PMID- 19541601 TI - Non-blinking and photostable upconverted luminescence from single lanthanide doped nanocrystals. AB - The development of probes for single-molecule imaging has dramatically facilitated the study of individual molecules in cells and other complex environments. Single-molecule probes ideally exhibit good brightness, uninterrupted emission, resistance to photobleaching, and minimal spectral overlap with cellular autofluorescence. However, most single-molecule probes are imperfect in several of these aspects, and none have been shown to possess all of these characteristics. Here we show that individual lanthanide-doped upconverting nanoparticles (UCNPs)--specifically, hexagonal phase NaYF(4) (beta-NaYF(4)) nanocrystals with multiple Yb(3+) and Er(3+) dopants--emit bright anti-Stokes visible upconverted luminescence with exceptional photostability when excited by a 980-nm continuous wave laser. Individual UCNPs exhibit no on/off emission behavior, or "blinking," down to the millisecond timescale, and no loss of intensity following an hour of continuous excitation. Amphiphilic polymer coatings permit the transfer of hydrophobic UCNPs into water, resulting in individual water-soluble nanoparticles with undiminished photophysical characteristics. These UCNPs are endocytosed by cells and show strong upconverted luminescence, with no measurable anti-Stokes background autofluorescence, suggesting that UCNPs are ideally suited for single-molecule imaging experiments. PMID- 19541602 TI - Influence of the coupling of interdomain and overall motions on NMR relaxation. AB - Most theoretical models for NMR relaxation in liquids assume that overall rotational motion can be described as rotational diffusion with a single diffusion tensor. Such models cannot handle motions (such as between "closed" and "open" states of an enzyme, or between conformers of a partially disordered system) where the shape of the molecule (and hence its rotational diffusion behavior) fluctuates. We provide here a formalism for dealing with such problems. The model involves jumps between discrete conformers with different overall diffusion tensors, and a master (rate) equation to describe the transitions between these conformers. Numerical examples are given for a two-site jump model where global and local motions are concerted, showing how the rate of conformational transitions (relative to the rate of rotational diffusion) affects the observed relaxation parameters. PMID- 19541603 TI - Positive selection optimizes the number and function of MHCII-restricted CD4+ T cell clones in the naive polyclonal repertoire. AB - T cell receptors (TCRs) on T lymphocytes in an individual bind foreign peptides bound to major histocompatibility complex (MHC) molecules expressed in that individual (designated MHC(A)). Results from radiation bone marrow chimeras and TCR transgenic mice indicate that this complex form of antigen recognition is the result of positive selection of clones with low affinity for self peptide:MHC(A) complexes during development. Here we used a sensitive peptide:MHC tetramer enrichment method to quantify the role of positive selection in the generation of the preimmune polyclonal T cell repertoire in normal individuals. We made the surprising observation that mouse and human naive T cells capable of binding to foreign peptide:MHC(A) were present at the same frequency in hosts that expressed MHC(A) or a different MHC isoform (MHC(B)). However, most of the clones in MHC(B) hosts also recognized self peptide:MHC(A) complexes. When these "alloreactive" T cells were removed from the MHC(B) repertoire via negative selection in an MHC(A) host, the number of foreign peptide:MHC(A)-binding T cells was reduced to one fifth and many of the remaining cells did not respond to the peptide. Therefore, although positive selection on MHC(A) was not required to produce foreign peptide:MHC(A)-binding clones, it had a large effect on selecting responsive clones. PMID- 19541604 TI - A metastable DWARF1 epigenetic mutant affecting plant stature in rice. AB - Epigenetic mutations confer heritable changes in gene expression that are not due to changes in the underlying sequence of the DNA. We identified a spontaneous rice mutant, Epi-d1, that shows a metastable dwarf phenotype. The phenotype is mitotically and meiotically inheritable and corresponds to the metastable epigenetic silencing of the DWARF1 (D1) gene. The silenced state is correlated with repressive histone and DNA methylation marks in the D1 promoter region but is not associated with DNA sequence alterations. Compared with other known epigenetic silenced loci in plants such as paramutable maize alleles and silent Arabidopsis genes, the Epi-d1 silencing phenomenon shows a high level of bidirectional metastable mutability. Epigenetic alleles such as Epi-d1 could thus provide for rapid adaptation under selective conditions. PMID- 19541605 TI - SPLUNC1 regulates airway surface liquid volume by protecting ENaC from proteolytic cleavage. AB - Many epithelia, including the superficial epithelia of the airways, are thought to secrete "volume sensors," which regulate the volume of the mucosal lining fluid. The epithelial Na(+) channel (ENaC) is often the rate limiting factor in fluid absorption, and must be cleaved by extracellular and/or intracellular proteases before it can conduct Na(+) and absorb excess mucosal liquid, a process that can be blocked by proteases inhibitors. In the airways, airway surface liquid dilution or removal activates ENaC. Therefore, we hypothesized that endogenous proteases are membrane-anchored, whereas endogenous proteolysis inhibitors are soluble and can function as airway surface liquid volume sensors to inhibit ENaC activity. Using a proteomic approach, we identified short palate, lung, and nasal epithelial clone (SPLUNC)1 as a candidate volume sensor. Recombinant SPLUNC1 inhibited ENaC activity in both human bronchial epithelial cultures and Xenopus oocytes. Knockdown of SPLUNC1 by shRNA resulted in a failure of bronchial epithelial cultures to regulate ENaC activity and airway surface liquid volume, which was restored by adding recombinant SPLUNC1 to the airway surface liquid. Despite being able to inhibit ENaC, recombinant SPLUNC1 had little effect on extracellular serine protease activity. However, SPLUNC1 specifically bound to ENaC, preventing its cleavage and activation by serine proteases. SPLUNC1 is highly expressed in the airways, as well as in colon and kidney. Thus, we propose that SPLUNC1 is secreted onto mucosal surfaces as a soluble volume sensor whose concentration and dilution can regulate ENaC activity and mucosal volumes, including that of airway surface liquid. PMID- 19541606 TI - Bayesian model predicts the response of axons to molecular gradients. AB - Axon guidance by molecular gradients plays a crucial role in wiring up the nervous system. However, the mechanisms axons use to detect gradients are largely unknown. We first develop a Bayesian "ideal observer" analysis of gradient detection by axons, based on the hypothesis that a principal constraint on gradient detection is intrinsic receptor binding noise. Second, from this model, we derive an equation predicting how the degree of response of an axon to a gradient should vary with gradient steepness and absolute concentration. Third, we confirm this prediction quantitatively by performing the first systematic experimental analysis of how axonal response varies with both these quantities. These experiments demonstrate a degree of sensitivity much higher than previously reported for any chemotacting system. Together, these results reveal both the quantitative constraints that must be satisfied for effective axonal guidance and the computational principles that may be used by the underlying signal transduction pathways, and allow predictions for the degree of response of axons to gradients in a wide variety of in vivo and in vitro settings. PMID- 19541607 TI - Transcription coactivator SAYP combines chromatin remodeler Brahma and transcription initiation factor TFIID into a single supercomplex. AB - Transcription activation by RNA polymerase II is a complicated process driven by combined, precisely coordinated action of a wide array of coactivator complexes, which carry out chromatin-directed activities and nucleate the assembly of the preinitiation complex on the promoter. Using various techniques, we have shown the existence of a stable coactivator supercomplex consisting of the chromatin remodeling factor Brahma (SWI/SNF) and the transcription initiation factor TFIID, named BTFly (Brahma and TFIID in one assembly). The coupling of Brahma and TFIID is mediated by the SAYP factor, whose evolutionarily conserved activation domain SAY can directly bind to both BAP170 subunit of Brahma and TAF5 subunit of TFIID. The integrity of BTFly is crucial for its ability to activate transcription. BTFly is distributed genome-wide and appears to be a means of effective transcription activation. PMID- 19541608 TI - Protein tyrosine phosphatase activity in the neural crest is essential for normal heart and skull development. AB - Mutations within the protein tyrosine phosphatase, SHP2, which is encoded by PTPN11, cause a significant proportion of Noonan syndrome (NS) cases, typically presenting with both cardiac disease and craniofacial abnormalities. Neural crest cells (NCCs) participate in both heart and skull formation, but the role of SHP2 signaling in NCC has not yet been determined. To gain insight into the role of SHP2 in NCC function, we ablated PTPN11 specifically in premigratory NCCs. SHP2 deficient NCCs initially exhibited normal migratory and proliferative patterns, but in the developing heart failed to migrate into the developing outflow tract. The embryos displayed persistent truncus arteriosus and abnormalities of the great vessels. The craniofacial deficits were even more pronounced, with large portions of the face and cranium affected, including the mandible and frontal and nasal bones. The data show that SHP2 activity in the NCC is essential for normal migration and differentiation into the diverse lineages found in the heart and skull and demonstrate the importance of NCC-based normal SHP2 activity in both heart and skull development, providing insight into the syndromic presentation characteristic of NS. PMID- 19541609 TI - mTOR and HIF-1alpha-mediated tumor metabolism in an LKB1 mouse model of Peutz Jeghers syndrome. AB - Peutz-Jeghers syndrome (PJS) is a familial cancer disorder due to inherited loss of function mutations in the LKB1/ STK11 serine/threonine kinase. PJS patients develop gastrointestinal hamartomas with 100% penetrance often in the second decade of life, and demonstrate an increased predisposition toward the development of a number of additional malignancies. Among mitogenic signaling pathways, the mammalian-target of rapamycin complex 1 (mTORC1) pathway is hyperactivated in tissues and tumors derived from LKB1-deficient mice. Consistent with a central role for mTORC1 in these tumors, rapamycin as a single agent results in a dramatic suppression of preexisting GI polyps in LKB1+/- mice. However, the key targets of mTORC1 in LKB1-deficient tumors remain unknown. We demonstrate here that these polyps, and LKB1- and AMPK-deficient mouse embryonic fibroblasts, show dramatic up-regulation of the HIF-1alpha transcription factor and its downstream transcriptional targets in an rapamycin-suppressible manner. The HIF-1alpha targets hexokinase II and Glut1 are up-regulated in these polyps, and using FDG-PET, we demonstrate that LKB1+/- mice show increased glucose utilization in focal regions of their GI tract corresponding to these gastrointestinal hamartomas. Importantly, we demonstrate that polyps from human Peutz-Jeghers patients similarly exhibit up-regulated mTORC1 signaling, HIF 1alpha, and GLUT1 levels. Furthermore, like HIF-1alpha and its target genes, the FDG-PET signal in the GI tract of these mice is abolished by rapamycin treatment. These findings suggest a number of therapeutic modalities for the treatment and detection of hamartomas in PJS patients, and potential for the screening and treatment of the 30% of sporadic human lung cancers bearing LKB1 mutations. PMID- 19541610 TI - Crystal structure of type I ryanodine receptor amino-terminal beta-trefoil domain reveals a disease-associated mutation "hot spot" loop. AB - Muscle contraction and relaxation is regulated by transient elevations of myoplasmic Ca(2+). Ca(2+) is released from stores in the lumen of the sarco(endo)plasmic reticulum (SER) to initiate formation of the Ca(2+) transient by activation of a class of Ca(2+) release channels referred to as ryanodine receptors (RyRs) and is pumped back into the SER lumen by Ca(2+)-ATPases (SERCAs) to terminate the Ca(2+) transient. Mutations in the type 1 ryanodine receptor gene, RYR1, are associated with 2 skeletal muscle disorders, malignant hyperthermia (MH), and central core disease (CCD). The evaluation of proposed mechanisms by which RyR1 mutations cause MH and CCD is hindered by the lack of high-resolution structural information. Here, we report the crystal structure of the N-terminal 210 residues of RyR1 (RyR(NTD)) at 2.5 A. The RyR(NTD) structure is similar to that of the suppressor domain of type 1 inositol 1,4,5 trisphosphate receptor (IP(3)Rsup), but lacks most of the long helix-turn-helix segment of the "arm" domain in IP(3)Rsup. The N-terminal beta-trefoil fold, found in both RyR and IP(3)R, is likely to play a critical role in regulatory mechanisms in this channel family. A disease-associated mutation "hot spot" loop was identified between strands 8 and 9 in a highly basic region of RyR1. Biophysical studies showed that 3 MH-associated mutations (C36R, R164C, and R178C) do not adversely affect the global stability or fold of RyR(NTD), supporting previously described mechanisms whereby mutations perturb protein protein interactions. PMID- 19541611 TI - Phase matching of high harmonic generation in the soft and hard X-ray regions of the spectrum. AB - We show how bright, tabletop, fully coherent hard X-ray beams can be generated through nonlinear upconversion of femtosecond laser light. By driving the high order harmonic generation process using longer-wavelength midinfrared light, we show that, in theory, fully phase-matched frequency upconversion can extend into the hard X-ray region of the spectrum. We verify our scaling predictions experimentally by demonstrating phase matching in the soft X-ray region of the spectrum around 330 eV, using ultrafast driving laser pulses at 1.3-microm wavelength, in an extended, high-pressure, weakly ionized gas medium. We also show through calculations that scaling of the overall conversion efficiency is surprisingly favorable as the wavelength of the driving laser is increased, making tabletop, fully coherent, multi-keV X-ray sources feasible. The rapidly decreasing microscopic single-atom yield, predicted for harmonics driven by longer-wavelength lasers, is compensated macroscopically by an increased optimal pressure for phase matching and a rapidly decreasing reabsorption of the generated X-rays. PMID- 19541612 TI - Zinc is an essential trace element for spermatogenesis. AB - Zinc (Zn) plays important roles in various biological activities but there is little available information regarding its functions in spermatogenesis. In our current study, we further examined the role of Zn during spermatogenesis in the Japanese eel (Anguilla japonica). Human CG (hCG) was injected into the animals to induce spermatogenesis, after which the concentration of Zn in the testis increased in tandem with the progression of spermatogenesis. Staining of testicular cells with a Zn-specific fluorescent probe revealed that Zn accumulates in germ cells, particularly in the mitochondria of spermatogonia and spermatozoa. Using an in vitro testicular organ culture system for the Japanese eel, production of a Zn deficiency by chelation with N,N,N',N'-tetrakis (2 pyridylemethyl)ethylenediamine (TPEN) caused apoptosis of the germ cells. However, this cell death was rescued by the addition of Zn to the cultures. Furthermore, an induced deficiency of Zn by TPEN chelation was found to inhibit the germ cell proliferation induced by 11-ketotestosterone (KT), a fish specific androgen, 17alpha,20beta-dihydroxy-4-pregnen-3-one (DHP), the initiator of meiosis in fish, and estradiol-17beta (E2), an inducer of spermatogonial stem cell renewal. We also investigated the effects of Zn deficiency on sperm motility and observed that TPEN treatment of eel sperm suppressed the rate and duration of their motility but that co-treatment with Zn blocked the effects of TPEN. Our present results thus suggest that Zn is an essential trace element for the maintenance of germ cells, the progression spermatogenesis, and the regulation of sperm motility. PMID- 19541613 TI - Biodesign of a renal-protective peptide based on alternative splicing of B-type natriuretic peptide. AB - Alternative RNA splicing may provide unique opportunities to identify drug targets and therapeutics. We identified an alternative spliced transcript for B type natriuretic peptide (BNP) resulting from intronic retention. This transcript is present in failing human hearts and is reduced following mechanical unloading. The intron-retained transcript would generate a unique 34 amino acid (aa) carboxyl terminus while maintaining the remaining structure of native BNP. We generated antisera to this carboxyl terminus and identified immunoreactivity in failing human heart tissue. The alternatively spliced peptide (ASBNP) was synthesized and unlike BNP, failed to stimulate cGMP in vascular cells or vasorelax preconstricted arterial rings. This suggests that ASBNP may lack the dose-limiting effects of recombinant BNP. Given structural considerations, a carboxyl-terminal truncated form of ASBNP was generated (ASBNP.1) and was determined to retain the ability of BNP to stimulate cGMP in canine glomerular isolates and cultured human mesangial cells but lacked similar effects in vascular cells. In a canine-pacing model of heart failure, systemic infusion of ASBNP.1 did not alter mean arterial pressure but increased the glomerular filtration rate (GFR), suppressed plasma renin and angiotensin, while inducing natriuresis and diuresis. Consistent with its distinct in vivo effects, the activity of ASBNP.1 may not be explained through binding and activation of NPR-A or NPR-B. Thus, the biodesigner peptide ASBNP.1 enhances GFR associated with heart failure while lacking the vasoactive properties of BNP. These findings demonstrate that peptides with unique properties may be designed based on products of alternatively splicing. PMID- 19541615 TI - Oviposition preference for and positional avoidance of acetic acid provide a model for competing behavioral drives in Drosophila. AB - Selection of appropriate oviposition sites is essential for progeny survival and fitness in generalist insect species, such as Drosophila melanogaster, yet little is known about the mechanisms regulating how environmental conditions and innate adult preferences are evaluated and balanced to yield the final substrate choice for egg-deposition. Female D. melanogaster are attracted to food containing acetic acid (AA) as an oviposition substrate. However, our observations reveal that this egg-laying preference is a complex process, as it directly opposes an otherwise strong, default behavior of positional avoidance for the same food. We show that 2 distinct sensory modalities detect AA. Attraction to AA-containing food for the purpose of egg-laying relies on the gustatory system, while positional repulsion depends primarily on the olfactory system. Similarly, distinct central brain regions are involved in AA attraction and repulsion. Given this unique situation, in which a single environmental stimulus yields 2 opposing behavioral outputs, we propose that the interaction of egg-laying attraction and positional aversion for AA provides a powerful model for studying how organisms balance competing behavioral drives and integrate signals involved in choice-like processes. PMID- 19541614 TI - Evaluating beta-turn mimics as beta-sheet folding nucleators. AB - Beta-turns are common conformations that enable proteins to adopt globular structures, and their formation is often rate limiting for folding. Beta-turn mimics, molecules that replace the i + 1 and i + 2 amino acid residues of a beta turn, are envisioned to act as folding nucleators by preorganizing the pendant polypeptide chains, thereby lowering the activation barrier for beta-sheet formation. However, the crucial kinetic experiments to demonstrate that beta-turn mimics can act as strong nucleators in the context of a cooperatively folding protein have not been reported. We have incorporated 6 beta-turn mimics simulating varied beta-turn types in place of 2 residues in an engineered beta turn 1 or beta-bulge turn 1 of the Pin 1 WW domain, a three-stranded beta-sheet protein. We present 2 lines of kinetic evidence that the inclusion of beta-turn mimics alters beta-sheet folding rates, enabling us to classify beta-turn mimics into 3 categories: those that are weak nucleators but permit Pin WW folding, native-like nucleators, and strong nucleators. Strong nucleators accelerate folding relative to WW domains incorporating all alpha-amino acid sequences. A solution NMR structure reveals that the native Pin WW beta-sheet structure is retained upon incorporating a strong E-olefin nucleator. These beta-turn mimics can now be used to interrogate protein folding transition state structures and the 2 kinetic analyses presented can be used to assess the nucleation capacity of other beta-turn mimics. PMID- 19541616 TI - Traceless protein splicing utilizing evolved split inteins. AB - Split inteins are parasitic genetic elements frequently found inserted into reading frames of essential proteins. Their association and excision restore host protein function through a protein self-splicing reaction. They have gained an increasingly important role in the chemical modification of proteins to create cyclical, segmentally labeled, and fluorescently tagged proteins. Ideally, inteins would seamlessly splice polypeptides together with no remnant sequences and at high efficiency. Here, we describe experiments that identify the branched intermediate, a transient step in the overall splicing reaction, as a key determinant of the splicing efficiency at different splice-site junctions. To alter intein specificity, we developed a cell-based selection scheme to evolve split inteins that splice with high efficiency at different splice junctions and at higher temperatures. Mutations within these evolved inteins occur at sites distant from the active site. We present a hypothesis that a network of conserved coevolving amino acids in inteins mediates these long-range effects. PMID- 19541617 TI - Nature of the protein universe. AB - The protein universe is the set of all proteins of all organisms. Here, all currently known sequences are analyzed in terms of families that have single domain or multidomain architectures and whether they have a known three dimensional structure. Growth of new single-domain families is very slow: Almost all growth comes from new multidomain architectures that are combinations of domains characterized by approximately 15,000 sequence profiles. Single-domain families are mostly shared by the major groups of organisms, whereas multidomain architectures are specific and account for species diversity. There are known structures for a quarter of the single-domain families, and >70% of all sequences can be partially modeled thanks to their membership in these families. PMID- 19541618 TI - Signaling dynamics of the KSR1 scaffold complex. AB - Scaffold proteins contribute to the spatiotemporal control of MAPK signaling and KSR1 is an ERK cascade scaffold that localizes to the plasma membrane in response to growth factor treatment. To better understand the molecular mechanisms of KSR1 function, we examined the interaction of KSR1 with each of the ERK cascade components, Raf, MEK, and ERK. Here, we identify a hydrophobic motif within the proline-rich sequence (PRS) of MEK1 and MEK2 that is required for constitutive binding to KSR1 and find that MEK binding and residues in the KSR1 CA1 region enable KSR1 to form a ternary complex with B-Raf and MEK following growth factor treatment that enhances MEK activation. We also find that docking of active ERK to the KSR1 scaffold allows ERK to phosphorylate KSR1 and B-Raf on feedback S/TP sites. Strikingly, feedback phosphorylation of KSR1 and B-Raf promote their dissociation and result in the release of KSR1 from the plasma membrane. Together, these findings provide unique insight into the signaling dynamics of the KSR1 scaffold and reveal that through regulated interactions with Raf and ERK, KSR1 acts to both potentiate and attenuate ERK cascade activation, thus regulating the intensity and duration of ERK cascade signaling emanating from the plasma membrane during growth factor signaling. PMID- 19541619 TI - An icosahedral algal virus has a complex unique vertex decorated by a spike. AB - Paramecium bursaria Chlorella virus-1 is an icosahedrally shaped, 1,900-A diameter virus that infects unicellular eukaryotic green algae. A 5-fold symmetric, 3D reconstruction using cryoelectron microscopy images has now shown that the quasiicosahedral virus has a unique vertex, with a pocket on the inside and a spike structure on the outside of the capsid. The pocket might contain enzymes for use in the initial stages of infection. The unique vertex consists of virally coded proteins, some of which have been identified. Comparison of shape, size, and location of the spike with similar features in bacteriophages T4 and P22 suggests that the spike might be a cell-puncturing device. Similar asymmetric features may have been missed in previous analyses of many other viruses that had been assumed to be perfectly icosahedral. PMID- 19541620 TI - The endocannabinoid system and pivotal role of the CB2 receptor in mouse spermatogenesis. AB - The exact role of the endocannabinoid system (ECS) during spermatogenesis has not been clarified. We used purified germ cell fractions representative of all phases of spermatogenesis and primary cultures of spermatogonia. This approach allowed the precise quantification of the cannabinoid receptor ligands, anandamide and 2 arachidonoylglycerol, and of the expression at transcriptional and transductional levels of their metabolic enzymes and receptors. Our data indicate that male mouse germ cells possess an active and complete ECS, which is modulated during meiosis, and suggest the presence of an autocrine endocannabinoid signal during spermatogenesis. Mitotic cells possess higher levels of 2-arachidonoylglycerol, which decrease in spermatocytes and spermatids. Accordingly, spermatogonia express higher and lower levels of 2-arachidonoylglycerol biosynthetic and degrading enzymes, respectively, as compared to meiotic and postmeiotic cells. This endocannabinoid likely plays a pivotal role in promoting the meiotic progression of germ cells by activating CB(2) receptors. In fact, we found that the selective CB(2) receptor agonist, JWH133, induced the Erk 1/2 MAPK phosphorylation cascade in spermatogonia and their progression toward meiosis, because it increased the number of cells positive for SCP3, a marker of meiotic prophase, and the expression of early meiotic prophase genes. PMID- 19541621 TI - Stress increases putative gonadotropin inhibitory hormone and decreases luteinizing hormone in male rats. AB - The subjective experience of stress leads to reproductive dysfunction in many species, including rodents and humans. Stress effects on reproduction result from multilevel interactions between the hormonal stress response system, i.e., the hypothalamic-pituitary-adrenal (HPA) axis, and the hormonal reproductive system, i.e., the hypothalamic-pituitary-gonadal (HPG) axis. A novel negative regulator of the HPG axis known as gonadotropin-inhibitory hormone (GnIH) was recently discovered in quail, and orthologous neuropeptides known as RFamide-related peptides (RFRPs) have also been identified in rodents and primates. It is currently unknown, however, whether GnIH/RFRPs influence HPG axis activity in response to stress. We show here that both acute and chronic immobilization stress lead to an up-regulation of RFRP expression in the dorsomedial hypothalamus (DMH) of adult male rats and that this increase in RFRP is associated with inhibition of downstream HPG activity. We also show that adrenalectomy blocks the stress-induced increase in RFRP expression. Immunohistochemistry revealed that 53% of RFRP cells express receptors for glucocorticoids (GCs), indicating that adrenal GCs can mediate the stress effect through direct action on RFRP cells. It is thought that stress effects on central control of reproduction are largely mediated by direct or indirect effects on GnRH-secreting neurons. Our data show that stress-induced increases in adrenal GCs cause an increase in RFRP that contributes to hypothalamic suppression of reproductive function. This novel insight into HPA-HPG interaction provides a paradigm shift for work on stress-related reproductive dysfunction and infertility, and indicates that future work on stress and reproductive system interactions must include investigation of the role of GnIH/RFRP. PMID- 19541622 TI - Multiple pathways guide oxygen diffusion into flavoenzyme active sites. AB - Dioxygen (O(2)) and other gas molecules have a fundamental role in a variety of enzymatic reactions. However, it is only poorly understood which O(2) uptake mechanism enzymes employ to promote efficient catalysis and how general this is. We investigated O(2) diffusion pathways into monooxygenase and oxidase flavoenzymes, using an integrated computational and experimental approach. Enhanced-statistics molecular dynamics simulations reveal spontaneous protein guided O(2) diffusion from the bulk solvent to preorganized protein cavities. The predicted protein-guided diffusion paths and the importance of key cavity residues for oxygen diffusion were verified by combining site-directed mutagenesis, rapid kinetics experiments, and high-resolution X-ray structures. This study indicates that monooxygenase and oxidase flavoenzymes employ multiple funnel-shaped diffusion pathways to absorb O(2) from the solvent and direct it to the reacting C4a atom of the flavin cofactor. The difference in O(2) reactivity among dehydrogenases, monooxygenases, and oxidases ultimately resides in the fine modulation of the local environment embedding the reactive locus of the flavin. PMID- 19541623 TI - Tools for thought or thoughts for tools? PMID- 19541624 TI - O2 reduction by a functional heme/nonheme bis-iron NOR model complex. AB - O(2) reactivity of a functional NOR model is investigated by using electrochemistry and spectroscopy. The electrochemical measurements using interdigitated electrodes show very high selectivity for 4e O(2) reduction with minimal production of partially reduced oxygen species (PROS) under both fast and slow electron flux. Intermediates trapped at cryogenic temperatures and characterized by using resonance Raman spectroscopy under single-turnover conditions indicate that an initial bridging peroxide intermediate undergoes homolytic O--O bond cleavage generating a trans heme/nonheme bis-ferryl intermediate. This bis ferryl species can oxygenate 2 equivalents of a reactive substrate. PMID- 19541625 TI - DDB2 decides cell fate following DNA damage. AB - The xeroderma pigmentosum complementation group E (XP-E) gene product damaged-DNA binding protein 2 (DDB2) plays important roles in nucleotide excision repair (NER). Previously, we showed that DDB2 participates in NER by regulating the level of p21(Waf1/Cip1). Here we show that the p21(Waf1/Cip1) -regulatory function of DDB2 plays a central role in defining the response (apoptosis or arrest) to DNA damage. The DDB2-deficient cells are resistant to apoptosis in response to a variety of DNA-damaging agents, despite activation of p53 and the pro-apoptotic genes. Instead, these cells undergo cell cycle arrest. Also, the DDB2-deficient cells are resistant to E2F1-induced apoptosis. The resistance to apoptosis of the DDB2-deficient cells is caused by an increased accumulation of p21(Waf1/Cip1) after DNA damage. We provide evidence that DDB2 targets p21(Waf1/Cip1) for proteolysis. The resistance to apoptosis in DDB2-deficient cells also involves Mdm2 in a manner that is distinct from the p53-regulatory activity of Mdm2. Our results provide evidence for a new regulatory loop involving the NER protein DDB2, Mdm2, and p21(Waf1/Cip1) that is critical in deciding cell fate (apoptosis or arrest) upon DNA damage. PMID- 19541626 TI - Dynamic features of gene expression control by small regulatory RNAs. AB - Small regulatory RNAs (sRNAs) in eukaryotes and bacteria play an important role in the regulation of gene expression either by binding to regulatory proteins or directly to target mRNAs. Two of the best-characterized bacterial sRNAs, Spot42 and RyhB, form a complementary pair with the ribosome binding region of their target mRNAs, thereby inhibiting translation or promoting mRNA degradation. To investigate the steady-state and dynamic potential of such sRNAs, we examine the 2 key parameters characterizing sRNA regulation: the capacity to overexpress the sRNA relative to its target mRNA and the speed at which the target mRNA is irreversibly inactivated. We demonstrate different methods to determine these 2 key parameters, for Spot42 and RyhB, which combine biochemical and genetic experiments with computational analysis. We have developed a mathematical model that describes the functional properties of sRNAs with various characteristic parameters. We observed that Spot42 and RyhB function in distinctive parameter regimes, which result in divergent mechanisms. PMID- 19541627 TI - Microfabricated tissue gauges to measure and manipulate forces from 3D microtissues. AB - Physical forces generated by cells drive morphologic changes during development and can feedback to regulate cellular phenotypes. Because these phenomena typically occur within a 3-dimensional (3D) matrix in vivo, we used microelectromechanical systems (MEMS) technology to generate arrays of microtissues consisting of cells encapsulated within 3D micropatterned matrices. Microcantilevers were used to simultaneously constrain the remodeling of a collagen gel and to report forces generated during this process. By concurrently measuring forces and observing matrix remodeling at cellular length scales, we report an initial correlation and later decoupling between cellular contractile forces and changes in tissue morphology. Independently varying the mechanical stiffness of the cantilevers and collagen matrix revealed that cellular forces increased with boundary or matrix rigidity whereas levels of cytoskeletal and extracellular matrix (ECM) proteins correlated with levels of mechanical stress. By mapping these relationships between cellular and matrix mechanics, cellular forces, and protein expression onto a bio-chemo-mechanical model of microtissue contractility, we demonstrate how intratissue gradients of mechanical stress can emerge from collective cellular contractility and finally, how such gradients can be used to engineer protein composition and organization within a 3D tissue. Together, these findings highlight a complex and dynamic relationship between cellular forces, ECM remodeling, and cellular phenotype and describe a system to study and apply this relationship within engineered 3D microtissues. PMID- 19541628 TI - His protects heme as it crosses the membrane. PMID- 19541629 TI - Diminished sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) expression contributes to airway remodelling in bronchial asthma. AB - Phenotypic modulation of airway smooth muscle (ASM) is an important feature of airway remodeling in asthma that is characterized by enhanced proliferation and secretion of pro-inflammatory chemokines. These activities are regulated by the concentration of free Ca(2+) in the cytosol ([Ca(2+)](i)). A rise in [Ca(2+)](i) is normalized by rapid reuptake of Ca(2+) into sarcoplasmic reticulum (SR) stores by the sarco/endoplasmic reticulum Ca(2+) (SERCA) pump. We examined whether increased proliferative and secretory responses of ASM from asthmatics result from reduced SERCA expression. ASM cells were cultured from subjects with and without asthma. SERCA expression was evaluated by western blot, immunohistochemistry and real-time PCR. Changes in [Ca(2+)](i), cell spreading, cellular proliferation, and eotaxin-1 release were measured. Compared with control cells from healthy subjects, SERCA2 mRNA and protein expression was reduced in ASM cells from subjects with moderately severe asthma. SERCA2 expression was similarly reduced in ASM in vivo in subjects with moderate/severe asthma. Rises in [Ca(2+)](i) following cell surface receptor-induced SR activation, or inhibition of SERCA-mediated Ca(2+) re-uptake, were attenuated in ASM cells from asthmatics. Likewise, the return to baseline of [Ca](i) after stimulation by bradykinin was delayed by approximately 50% in ASM cells from asthmatics. siRNA-mediated knockdown of SERCA2 in ASM from healthy subjects increased cell spreading, eotaxin-1 release and proliferation. Our findings implicate a deficiency in SERCA2 in ASM in asthma that contributes to its secretory and hyperproliferative phenotype in asthma, and which may play a key role in mechanisms of airway remodeling. PMID- 19541630 TI - NOD2 contributes to cutaneous defense against Staphylococcus aureus through alpha toxin-dependent innate immune activation. AB - Staphylococcus aureus is a major cause of community-acquired and nosocomial infections including the life-threatening conditions endocarditis, necrotizing pneumonia, necrotizing fasciitis, and septicemia. Toll-like receptor (TLR)-2, a membrane-bound microbial sensor, detects staphylococcal components, but macrophages lacking TLR2 or both TLR2 and TLR4 remain S. aureus responsive, suggesting that an alternative microbial recognition receptor might be involved. The cytoplasmic sensor nucleotide-binding oligomerization domain containing (NOD) 2/caspase recruitment domain (CARD) 15 detects muramyl dipeptide from bacterial peptidoglycans and mediates cytokine responses to S. aureus in vitro, but the physiological significance of these observations is not well defined. Here we show that NOD2-deficient mice exhibit a delayed but ultimately exacerbated ulcerative response and impaired bacterial clearance after s.c. infection with S. aureus. NOD2-dependent recognition of S. aureus and muramyl dipeptide is facilitated by alpha-toxin (alpha-hemolysin), a pore-forming toxin and virulence factor of the pathogen. The action of NOD2 is dependent on IL-1beta-amplified production of IL-6, which promotes rapid bacterial killing by neutrophils. These results significantly broaden the physiological importance of NOD2 in innate immunity from the recognition of bacteria that primarily enter the cytoplasm to the detection of bacteria that typically reside extracellularly and demonstrate that this microbial sensor contributes to the discrimination between commensal bacteria and bacterial pathogens that elaborate pore-forming toxins. PMID- 19541631 TI - Real-time molecular monitoring of chemical environment in obligate anaerobes during oxygen adaptive response. AB - Determining the transient chemical properties of the intracellular environment can elucidate the paths through which a biological system adapts to changes in its environment, for example, the mechanisms that enable some obligate anaerobic bacteria to survive a sudden exposure to oxygen. Here we used high-resolution Fourier transform infrared (FTIR) spectromicroscopy to continuously follow cellular chemistry within living obligate anaerobes by monitoring hydrogen bond structures in their cellular water. We observed a sequence of well orchestrated molecular events that correspond to changes in cellular processes in those cells that survive, but only accumulation of radicals in those that do not. We thereby can interpret the adaptive response in terms of transient intracellular chemistry and link it to oxygen stress and survival. This ability to monitor chemical changes at the molecular level can yield important insights into a wide range of adaptive responses. PMID- 19541632 TI - The glucanosyltransferase Gas1 functions in transcriptional silencing. AB - Transcriptional silencing is a crucial process that is mediated through chromatin structure. The histone deacetylase Sir2 silences genomic regions that include telomeres, ribosomal DNA (rDNA) and the cryptic mating-type loci. Here, we report an unsuspected role for the enzyme Gas1 in locus-specific transcriptional silencing. GAS1 encodes a beta-1,3-glucanosyltransferase previously characterized for its role in cell wall biogenesis. In gas1 mutants, telomeric silencing is defective and rDNA silencing is enhanced. We show that the catalytic activity of Gas1 is required for normal silencing, and that Gas1's role in silencing is distinct from its role in cell wall biogenesis. Established hallmarks of silent chromatin, such as Sir2 and Sir3 binding, H4K16 deacetylation, and H3K56 deacetylation, appear unaffected in gas1 mutants. Thus, another event required for telomeric silencing must be influenced by GAS1. Because the catalytic activity of Gas1 is required for telomeric silencing, Gas1 localizes to the nuclear periphery, and Gas1 and Sir2 physically interact, we propose a model in which carbohydrate modification of chromatin components provides a new regulatory element that may be critical for chromatin function but which is virtually unexplored in the current landscape of chromatin analysis. PMID- 19541633 TI - Osmolyte-induced separation of the mechanical folding phases of ubiquitin. AB - Solvent molecules play key roles in the conformational dynamics of proteins. Here we use single molecule force-clamp spectroscopy to probe the role played by the stabilizing osmolyte glycerol on the conformational ensembles visited by a single ubiquitin protein folding after mechanical extension. Using a variety of force pulse protocols, we find that glycerol stabilizes the native state of ubiquitin, making it more resistant to mechanical unfolding. We also find that although glycerol enhanced the hydrophobic collapse of unfolded and highly extended ubiquitins, it had no effect on the resulting collapsed states that are essential precursors of the folded state. These disparate effects of glycerol may be the result of distinct structural roles played by solvent molecules at the transition state of each folding ensemble. Our results open the way for a detailed analysis of the transition state structures that form along the folding trajectory of a mechanically extended protein. PMID- 19541634 TI - Functional analysis of an Orc6 mutant in Drosophila. AB - The origin recognition complex (ORC) is a 6-subunit complex required for the initiation of DNA replication in eukaryotic organisms. ORC is also involved in other cell functions. The smallest Drosophila ORC subunit, Orc6, is important for both DNA replication and cytokinesis. To study the role of Orc6 in vivo, the orc6 gene was deleted by imprecise excision of P element. Lethal alleles of orc6 are defective in DNA replication and also show abnormal chromosome condensation and segregation. The analysis of cells containing the orc6 deletion revealed that they arrest in both the G(1) and mitotic stages of the cell cycle. Orc6 deletion can be rescued to viability by a full-length Orc6 transgene. The expression of mutant transgenes of Orc6 with deleted or mutated C-terminal domain results in a release of mutant cells from G(1) arrest and restoration of DNA replication, indicating that the DNA replication function of Orc6 is associated with its N terminal domain. However, these mutant cells accumulate at mitosis, suggesting that the C-terminal domain of Orc6 is important for the passage through the M phase. In a cross-species complementation experiment, the expression of human Orc6 in Drosophila Orc6 mutant cells rescued DNA replication, suggesting that this function of the protein is conserved among metazoans. PMID- 19541635 TI - Direct observation of an ensemble of stable collapsed states in the mechanical folding of ubiquitin. AB - Statistical theories of protein folding have long predicted plausible mechanisms for reducing the vast conformational space through distinct ensembles of structures. However, these predictions have remained untested by bulk techniques, because the conformational diversity of folding molecules has been experimentally unapproachable. Owing to recent advances in single molecule force-clamp spectroscopy, we are now able to probe the structure and dynamics of the small protein ubiquitin by measuring its length and mechanical stability during each stage of folding. Here, we discover that upon hydrophobic collapse, the protein rapidly selects a subset of minimum energy structures that are mechanically weak and essential precursors of the native fold. From this much reduced ensemble, the native state is acquired through a barrier-limited transition. Our results support the validity of statistical mechanics models in describing the folding of a small protein on biological timescales. PMID- 19541636 TI - Sodium-calcium exchanger complexed with GM1 ganglioside in nuclear membrane transfers calcium from nucleoplasm to endoplasmic reticulum. AB - The inner membrane of the nuclear envelope (NE) was previously shown to contain a Na/Ca exchanger (NCX) tightly linked to GM1 ganglioside that mediates transfer of nucleoplasmic Ca(2+) to the NE lumen and constitutes a cytoprotective mechanism. This transfer was initially observed with isolated nuclei and is now demonstrated in living cells in relation to subcellular Ca(2+) dynamics. Four cell lines with varying expression of NCX and GM1 in the NE were transfected with cameleon fluorescent Ca(2+) indicators genetically targeted to NE/endoplasmic reticulum (ER) and nucleoplasm to monitor [Ca(2+)](ne/er) and [Ca(2+)](n) respectively. Cytosolic Ca(2+) ([Ca(2+)](cyt)) was indicated with fura-2. Thapsigargin caused progressive loss of [Ca(2+)](ne/er), which was rapidly replaced on addition of extrinsic Ca(2+) to those cells containing fully functional NCX/GM1: differentiated NG108-15 and C6 cells. Reduced elevation of [Ca(2+)](ne/er) following thapsigargin depletion occurred in cells containing little or no GM1 in the NE: undifferentiated NG108-15 and NG-CR72 cells. No change in [Ca(2+)](ne/er) due to applied Ca(2+) was seen in Jurkat cells, which entirely lack NCX. Ca(2+) entry to NE/ER was also blocked by KB-R7943, inhibitor of NCX. [Ca(2+)](n) and [Ca(2+)](cyt) were elevated independent of [Ca(2+)](ne/er) and remained in approximate equilibrium with each other. Ca(2+) rise in the ER originated in the NE region and extended to the entire ER network. These results indicate the nuclear NCX/GM1 complex acts to gate Ca(2+) transfer from cytosol to ER, an alternate route to the sarcoplasmic/endoplasmic reticulum calcium ATPase pump. They also suggest a possible contributory mechanism for independent regulation of nuclear Ca(2+). PMID- 19541637 TI - Sporulation in mycobacteria. AB - Mycobacteria owe their success as pathogens to their ability to persist for long periods within host cells in asymptomatic, latent forms before they opportunistically switch to the virulent state. The molecular mechanisms underlying the transition into dormancy and emergence from it are not clear. Here we show that old cultures of Mycobacterium marinum contained spores that, upon exposure to fresh medium, germinated into vegetative cells and reappeared again in stationary phase via endospore formation. They showed many of the usual characteristics of well-known endospores. Homologues of well-known sporulation genes of Bacillus subtilis and Streptomyces coelicolor were detected in mycobacteria genomes, some of which were verified to be transcribed during appropriate life-cycle stages. We also provide data indicating that it is likely that old Mycobacterium bovis bacillus Calmette-Guerin cultures form spores. Together, our data show sporulation as a lifestyle adapted by mycobacteria under stress and tempt us to suggest this as a possible mechanism for dormancy and/or persistent infection. If so, this might lead to new prophylactic strategies. PMID- 19541639 TI - Temporal lenses for attosecond and femtosecond electron pulses. AB - Here, we describe the "temporal lens" concept that can be used for the focus and magnification of ultrashort electron packets in the time domain. The temporal lenses are created by appropriately synthesizing optical pulses that interact with electrons through the ponderomotive force. With such an arrangement, a temporal lens equation with a form identical to that of conventional light optics is derived. The analog of ray diagrams, but for electrons, are constructed to help the visualization of the process of compressing electron packets. It is shown that such temporal lenses not only compensate for electron pulse broadening due to velocity dispersion but also allow compression of the packets to durations much shorter than their initial widths. With these capabilities, ultrafast electron diffraction and microscopy can be extended to new domains,and, just as importantly, electron pulses can be delivered directly on an ultrafast techniques target specimen. PMID- 19541638 TI - NAADP-mediated Ca2+ signaling via type 1 ryanodine receptor in T cells revealed by a synthetic NAADP antagonist. AB - The nucleotide NAADP was recently discovered as a second messenger involved in the initiation and propagation of Ca(2+) signaling in lymphoma T cells, but its impact on primary T cell function is still unknown. An optimized, synthetic, small molecule inhibitor of NAADP action, termed BZ194, was designed and synthesized. BZ194 neither interfered with Ca(2+) mobilization by d-myo-inositol 1,4,5-trisphosphate or cyclic ADP-ribose nor with capacitative Ca(2+) entry. BZ194 specifically and effectively blocked NAADP-stimulated [(3)H]ryanodine binding to the purified type 1 ryanodine receptor. Further, in intact T cells, Ca(2+) mobilization evoked by NAADP or by formation of the immunological synapse between primary effector T cells and astrocytes was inhibited by BZ194. Downstream events of Ca(2+) mobilization, such as nuclear translocation of "nuclear factor of activated T cells" (NFAT), T cell receptor-driven interleukin 2 production, and proliferation in antigen-experienced CD4(+) effector T cells, were attenuated by the NAADP antagonist. Taken together, specific inhibition of the NAADP signaling pathway constitutes a way to specifically and effectively modulate T-cell activation and has potential in the therapy of autoimmune diseases. PMID- 19541640 TI - Unbiased RNA-protein interaction screen by quantitative proteomics. AB - Mass spectrometry (MS)-based quantitative interaction proteomics has successfully elucidated specific protein-protein, DNA-protein, and small molecule-protein interactions. Here, we developed a gel-free, sensitive, and scalable technology that addresses the important area of RNA-protein interactions. Using aptamer tagged RNA as bait, we captured RNA-interacting proteins from stable isotope labeling by amino acids in cell culture (SILAC)-labeled mammalian cell extracts and analyzed them by high-resolution, quantitative MS. Binders specific to the RNA sequence were distinguished from background by their isotope ratios between bait and control. We demonstrated the approach by retrieving known and novel interaction partners for the HuR interaction motif, H4 stem loop, "zipcode" sequence, tRNA, and a bioinformatically-predicted RNA fold in DGCR-8/Pasha mRNA. In all experiments we unambiguously identified known interaction partners by a single affinity purification step. The 5' region of the mRNA of DGCR-8/Pasha, a component of the microprocessor complex, specifically interacts with components of the translational machinery, suggesting that it contains an internal ribosome entry site. PMID- 19541641 TI - Top-down high-resolution mass spectrometry of cardiac myosin binding protein C revealed that truncation alters protein phosphorylation state. AB - Cardiac myosin binding protein C (cMyBP-C), bound to the sarcomere's myosin thick filament, plays an important role in the regulation of muscle contraction. cMyBP C is a large multidomain protein that interacts with myosin, titin, and possibly actin. Mutations in cMyBP-C are the most common known cause of heritable hypertrophic cardiomypathies. Phosphorylation of cMyBP-C plays an essential role in the normal cardiac function. cMyBP-C (142 kDa) has 81 serine and 73 threonine residues presenting a major challenge for unequivocal identification of specific phosphorylation sites. Top-down mass spectrometry, which directly analyzes intact proteins, is a powerful technique to universally observe and quantify protein posttranslational modifications without a priori knowledge. Here, we have extended top-down electron capture dissociation mass spectrometry to comprehensively characterize mouse cMyBP-C expressed in baculovirus. We have unambiguously identified all of the phosphorylation sites in the truncated (28 115 kDa) and full-length forms of cMyBP-C (142 kDa) and characterized the sequential phosphorylations, using a combination of top-down and middle-down (limited proteolysis) MS approach, which ensures full sequence coverage. Unit mass resolution and high mass accuracy (<5 ppm) have been achieved for a 115-kDa protein (the largest protein isotopically resolved to date). Remarkably, we discovered that truncations in recombinant proteins, even a seemingly minor one, can dramatically alter its phosphorylation state, which is significant because truncated recombinant proteins are routinely substituted for their full-length forms in crystal structure and functional studies. Our study provides direct evidence of alterations in the posttranslational state between the truncated and full-length recombinant proteins, which can lead to variations in structure and function. PMID- 19541643 TI - Melting and crystallization of colloidal hard-sphere suspensions under shear. AB - Shear-induced melting and crystallization were investigated by confocal microscopy in concentrated colloidal suspensions of hard-sphere-like particles. Both silica and polymethylmethacrylate suspensions were sheared with a constant rate in either a countertranslating parallel plate shear cell or a counterrotating cone-plate shear cell. These instruments make it possible to track particles undergoing shear for extended periods of time in a plane of zero velocity. Although on large scales, the flow profile deviated from linearity, the crystal flowed in an aligned sliding layer structure at low shear rates. Higher shear rates caused the crystal to shear melt, but, contrary to expectations, the transition was not sudden. Instead, although the overall order decreased with shear rate, this was due to an increase in the nucleation of localized domains that temporarily lost and regained their ordered structure. Even at shear rates that were considered to have melted the crystal as a whole, ordered regions kept showing up at times, giving rise to very large fluctuations in 2D bond orientational order parameters. Low shear rates induced initially disordered suspensions to crystallize. This time, the order parameter increased gradually in time without large fluctuations, indicating that shear-induced crystallization of hard spheres does not proceed via a nucleation and growth mechanism. We conclude that the dynamics of melting and crystallization under shear differ dramatically from their counterparts in quiescent suspensions. PMID- 19541642 TI - FGF21 induces PGC-1alpha and regulates carbohydrate and fatty acid metabolism during the adaptive starvation response. AB - The liver plays a crucial role in mobilizing energy during nutritional deprivation. During the early stages of fasting, hepatic glycogenolysis is a primary energy source. As fasting progresses and glycogen stores are depleted, hepatic gluconeogenesis and ketogenesis become major energy sources. Here, we show that fibroblast growth factor 21 (FGF21), a hormone that is induced in liver by fasting, induces hepatic expression of peroxisome proliferator-activated receptor gamma coactivator protein-1alpha (PGC-1alpha), a key transcriptional regulator of energy homeostasis, and causes corresponding increases in fatty acid oxidation, tricarboxylic acid cycle flux, and gluconeogenesis without increasing glycogenolysis. Mice lacking FGF21 fail to fully induce PGC-1alpha expression in response to a prolonged fast and have impaired gluconeogenesis and ketogenesis. These results reveal an unexpected relationship between FGF21 and PGC-1alpha and demonstrate an important role for FGF21 in coordinately regulating carbohydrate and fatty acid metabolism during the progression from fasting to starvation. PMID- 19541644 TI - Cellular immune response to cryptic epitopes during therapeutic gene transfer. AB - The immune response has been implicated as a critical factor in determining the success or failure of clinical gene therapy trials. Generally, such a response is elicited by the desired transgene product or, in some cases, the delivery system. In the current study, we report the previously uncharacterized finding that a therapeutic cassette currently being used for human investigation displays alternative reading frames (ARFs) that generate unwanted protein products to induce a cytotoxic T lymphocyte (CTL) response. In particular, we tested the hypothesis that antigenic epitopes derived from an ARF in coagulation factor IX (F9) cDNA can induce CTL reactivity, subsequently killing F9-expressing hepatocytes. One peptide (p18) of 3 candidates from an ARF of the F9 transgene induced CD8(+) T cell reactivity in mice expressing the human MHC class I molecule B0702. Subsequently, upon systemic administration of adeno-associated virus (AAV) serotype 2 vectors packaged with the F9 transgene (AAV2/F9), a robust CD8(+) CTL response was elicited against peptide p18. Of particular importance is that the ARF epitope-specific CTLs eliminated AAV2/F9-transduced hepatocytes but not AAV2/F9 codon-optimized (AAV2/F9-opt)-transduced liver cells in which p18 epitope was deleted. These results demonstrate a previously undiscovered mechanism by which CTL responses can be elicited by cryptic epitopes generated from a therapeutic transgene and have significant implications for all gene therapy modalities. Such unforeseen epitope generation warrants careful analysis of transgene sequences for ARFs to reduce the potential for adverse events arising from immune responses during clinical gene therapy protocols. PMID- 19541645 TI - Activation of tumor cell integrin alphavbeta3 controls angiogenesis and metastatic growth in the brain. AB - The incidence of brain metastasis is rising and poses a severe clinical problem, as we lack effective therapies and knowledge of mechanisms that control metastatic growth in the brain. Here we demonstrate a crucial role for high affinity tumor cell integrin alpha(v)beta(3) in brain metastatic growth and recruitment of blood vessels. Although alpha(v)beta(3) is frequently up-regulated in primary brain tumors and metastatic lesions of brain homing cancers, we show that it is the alpha(v)beta(3) activation state that is critical for brain lesion growth. Activated, but not non-activated, tumor cell alpha(v)beta(3) supports efficient brain metastatic growth through continuous up-regulation of vascular endothelial growth factor (VEGF) protein under normoxic conditions. In metastatic brain lesions carrying activated alpha(v)beta(3), VEGF expression is controlled at the post-transcriptional level and involves phosphorylation and inhibition of translational respressor 4E-binding protein (4E-BP1). In contrast, tumor cells with non-activated alpha(v)beta(3) depend on hypoxia for VEGF induction, resulting in reduced angiogenesis, tumor cell apoptosis, and inefficient intracranial growth. Importantly, the microenvironment critically influences the effects that activated tumor cell alpha(v)beta(3) exerts on tumor cell growth. Although it strongly promoted intracranial growth, the activation state of the receptor did not influence tumor growth in the mammary fat pad as a primary site. Thus, we identified a mechanism by which metastatic cells thrive in the brain microenvironment and use the high-affinity form of an adhesion receptor to grow and secure host support for proliferation. Targeting this molecular mechanism could prove valuable for the inhibition of brain metastasis. PMID- 19541646 TI - Mesoscopic atomic entanglement for precision measurements beyond the standard quantum limit. AB - Squeezing of quantum fluctuations by means of entanglement is a well-recognized goal in the field of quantum information science and precision measurements. In particular, squeezing the fluctuations via entanglement between 2-level atoms can improve the precision of sensing, clocks, metrology, and spectroscopy. Here, we demonstrate 3.4 dB of metrologically relevant squeezing and entanglement for greater, similar 10(5) cold caesium atoms via a quantum nondemolition (QND) measurement on the atom clock levels. We show that there is an optimal degree of decoherence induced by the quantum measurement which maximizes the generated entanglement. A 2-color QND scheme used in this paper is shown to have a number of advantages for entanglement generation as compared with a single-color QND measurement. PMID- 19541648 TI - Narrowing of intersensory speech perception in infancy. AB - The conventional view is that perceptual/cognitive development is an incremental process of acquisition. Several striking findings have revealed, however, that the sensitivity to non-native languages, faces, vocalizations, and music that is present early in life declines as infants acquire experience with native perceptual inputs. In the language domain, the decline in sensitivity is reflected in a process of perceptual narrowing that is thought to play a critical role during the acquisition of a native-language phonological system. Here, we provide evidence that such a decline also occurs in infant response to multisensory speech. We found that infant intersensory response to a non-native phonetic contrast narrows between 6 and 11 months of age, suggesting that the perceptual system becomes increasingly more tuned to key native-language audiovisual correspondences. Our findings lend support to the notion that perceptual narrowing is a domain-general as well as a pan-sensory developmental process. PMID- 19541647 TI - Computing protein stabilities from their chain lengths. AB - New amino acid sequences of proteins are being learned at a rapid rate, thanks to modern genomics. The native structures and functions of those proteins can often be inferred using bioinformatics methods. We show here that it is also possible to infer the stabilities and thermal folding properties of proteins, given only simple genomics information: the chain length and the numbers of charged side chains. In particular, our model predicts DeltaH(T), DeltaS(T), DeltaC(p), and DeltaF(T)--the folding enthalpy, entropy, heat capacity, and free energy--as functions of temperature T; the denaturant m values in guanidine and urea; the pH temperature-salt phase diagrams, and the energy of confinement F(s) of the protein inside a cavity of radius s. All combinations of these phase equilibria can also then be computed from that information. As one illustration, we compute the pH and salt conditions that would denature a protein inside a small confined cavity. Because the model is analytical, it is computationally efficient enough that it could be used to automatically annotate whole proteomes with protein stability information. PMID- 19541649 TI - Contingencies and compounded rare perturbations dictate sudden distributional shifts during periods of gradual climate change. AB - Ecological responses to climate change may occur gradually with changing conditions, or they may occur rapidly once some threshold or "tipping point" has been reached. Here, we use a high-resolution, 30-year data set on the upper vertical limit of a high intertidal alga to demonstrate that distributional shifts in this species do not keep pace with gradual trends in air temperature or sea level, but rather occur in sudden, discrete steps. These steps occur when unusually warm air temperatures are associated with unusually calm seas and are contingent in the sense that neither atmospheric nor sea conditions by themselves were sufficient to generate the underlying physiological challenge. Shifts in the upper limit did not correlate with large environmental perturbations such as El Ninos; rather, they appeared to be associated with stochastic departures from otherwise gradual environmental trends. Our results exemplify the view that multiple environmental factors should be considered when attempting to understand ecological responses to climate change. Furthermore, punctuated responses such as those we have identified urge caution when attempting to infer causal mechanisms and project future distributional shifts using data of limited temporal resolution or scope. PMID- 19541650 TI - Signaling mechanisms involved in altered function of macrophages from diet induced obese mice affect immune responses. AB - Recent research links diet-induced obesity (DIO) with impaired immunity, although the underlying mechanisms remain unclear. We find that the induction of inducible NO synthase (iNOS) and cytokines is suppressed in mice with DIO and in bone marrow macrophages (BMMPhi) from mice with DIO exposed to an oral pathogen, Porphyromonas gingivalis. BMMPhi from lean mice pre-treated with free fatty acids (FFAs) and exposed to P. gingivalis also exhibit a diminished induction of iNOS and cytokines. BMMPhi from lean and obese mice exposed to P. gingivalis and analyzed by a phosphorylation protein array show a reduction of Akt only in BMMPhi from mice with DIO. This reduction is responsible for diminished NF-kappaB activation and diminished induction of iNOS and cytokines. We next observed that Toll-like receptor 2 (TLR2) is suppressed in BMMPhi from DIO mice whereas carboxy terminal modulator protein (CTMP), a known suppressor of Akt phosphorylation, is elevated. This elevation stems from defective TLR2 signaling. In BMMPhi from lean mice, both FFAs and TNF-alpha--via separate pathways--induce an increase in CMTP. However, in BMMPhi from DIO mice, TLR2 can no longer inhibit the TNF-alpha induced increase in CTMP caused by P. gingivalis challenge. This defect can then be restored by transfecting WT TLR2 into BMMPhi from DIO mice. Thus, feeding mice a high-fat diet over time elevates the CTMP intracellular pool, initially via FFAs activating TLR2 and later when the defective TLR2 is unable to inhibit TNF alpha-induced CTMP. These findings unveil a link between obesity and innate immunity. PMID- 19541652 TI - Nanogranular origin of concrete creep. AB - Concrete, the solid that forms at room temperature from mixing Portland cement with water, sand, and aggregates, suffers from time-dependent deformation under load. This creep occurs at a rate that deteriorates the durability and truncates the lifespan of concrete structures. However, despite decades of research, the origin of concrete creep remains unknown. Here, we measure the in situ creep behavior of calcium-silicate-hydrates (C-S-H), the nano-meter sized particles that form the fundamental building block of Portland cement concrete. We show that C-S-H exhibits a logarithmic creep that depends only on the packing of 3 structurally distinct but compositionally similar C-S-H forms: low density, high density, ultra-high density. We demonstrate that the creep rate ( approximately 1/t) is likely due to the rearrangement of nanoscale particles around limit packing densities following the free-volume dynamics theory of granular physics. These findings could lead to a new basis for nanoengineering concrete materials and structures with minimal creep rates monitored by packing density distributions of nanoscale particles, and predicted by nanoscale creep measurements in some minute time, which are as exact as macroscopic creep tests carried out over years. PMID- 19541651 TI - Adenosine A2A receptor is a unique angiogenic target of HIF-2alpha in pulmonary endothelial cells. AB - Hypoxia, through the hypoxia-inducible transcription factors HIF-1alpha and HIF 2alpha (HIFs), induces angiogenesis by up-regulating a common set of angiogenic cytokines. Unlike HIF-1alpha, which regulates a unique set of genes, most genes regulated by HIF-2alpha overlap with those induced by HIF-1alpha. Thus, the unique contribution of HIF-2alpha remains largely obscure. By using adenoviral mutant HIF-1alpha and adenoviral mutant HIF-2alpha constructs, where the HIFs are transcriptionally active under normoxic conditions, we show that HIF-2alpha but not HIF-1alpha regulates adenosine A(2A) receptor in primary cultures of human lung endothelial cells. Further, siRNA knockdown of HIF-2alpha completely inhibits hypoxic induction of A(2A) receptor. Promoter studies show a 2.5-fold induction of luciferase activity with HIF-2alpha cotransfection. Analysis of the A(2A) receptor gene promoter revealed a hypoxia-responsive element in the region between -704 and -595 upstream of the transcription start site. By using a ChIP assay, we demonstrate that HIF-2alpha binding to this region is specific. In addition, we demonstrate that A(2A) receptor has angiogenic potential, as assessed by increases in cell proliferation, cell migration, and tube formation. Additional data show increased expression of A(2A) receptor in human lung tumor cancer samples relative to adjacent normal lung tissue. These data also demonstrate that A(2A) receptor is regulated by hypoxia and HIF-2alpha in human lung endothelial cells but not in mouse-derived endothelial cells. PMID- 19541653 TI - The double-stranded RNA-binding protein, PACT, is required for postnatal anterior pituitary proliferation. AB - PACT is a double-stranded RNA-binding protein that also binds and activates the latent protein kinase, PKR, which plays a major role in cellular antiviral defense in mammals. For evaluating PACT's contribution to the innate immune system, Pact(-/-) mice have been generated; these mice exhibit notable developmental abnormalities including microtia, with craniofacial, ear, and hearing defects. Here we report that, in addition, Pact(-/-) mice had smaller body size and fertility defects, both of which were caused by defective pituitary functions. Pact(-/-) mice exhibited anterior pituitary lobe (AL) hypoplasia, which developed postnatally, when the second phase of pituitary expansion occurs. Among the 5 cell types in AL, the numbers of corticotrophs, gonadotrophs, and somatotrophs were equally decreased in Pact(-/-) mice with a greater impact on lactotrophs and a lesser impact on thyrotrophs. PACT mRNA and protein were highly expressed in the pituitary of wild-type (Wt) mice during the postnatal wave of AL proliferation, the same period in which the hypoplasia developed in Pact(-/-) mice. During this time, the pituitaries of Pact(-/-) mice did not exhibit significantly increased apoptosis compared with Wt mice but showed a decrease in cell proliferation. The inhibition of cell proliferation observed in vivo could be recapitulated in vitro in GH3 somato/lactotroph and LbetaT2 gonadotroph cell lines; knockdown of PACT expression with siRNA diminished the rate of proliferation of these cells. Our study revealed a physiologically significant role for PACT in cell proliferation and an essential role of a dsRNA-binding protein in mammalian pituitary expansion. PMID- 19541654 TI - Structural and kinetic modeling of an activating helix switch in the rhodopsin transducin interface. AB - Extracellular signals prompt G protein-coupled receptors (GPCRs) to adopt an active conformation (R*) and catalyze GDP/GTP exchange in the alpha-subunit of intracellular G proteins (Galphabetagamma). Kinetic analysis of transducin (G(t)alphabetagamma) activation shows that an intermediary R*xG(t)alphabetagamma.GDP complex is formed that precedes GDP release and formation of the nucleotide-free R*xG protein complex. Based on this reaction sequence, we explore the dynamic interface between the proteins during formation of these complexes. We start from the R* conformation stabilized by a G(t)alpha C terminal peptide (GalphaCT) obtained from crystal structures of the GPCR opsin. Molecular modeling allows reconstruction of the fully elongated C-terminal alpha helix of G(t)alpha (alpha5) and shows how alpha5 can be docked to the open binding site of R*. Two modes of interaction are found. One of them--termed stable or S-interaction--matches the position of the GalphaCT peptide in the crystal structure and reproduces the hydrogen-bonding networks between the C terminal reverse turn of GalphaCT and conserved E(D)RY and NPxxY(x)(5,6)F regions of the GPCR. The alternative fit--termed intermediary or I-interaction--is distinguished by a tilt (42 degrees ) and rotation (90 degrees ) of alpha5 relative to the S-interaction and shows different alpha5 contacts with the NPxxY(x)(5,6)F region and the second cytoplasmic loop of R*. From the 2 alpha5 interactions, we derive a "helix switch" mechanism for the transition of R*xG(t)alphabetagamma.GDP to the nucleotide-free R*xG protein complex that illustrates how alpha5 might act as a transmission rod to propagate the conformational change from the receptor-G protein interface to the nucleotide binding site. PMID- 19541655 TI - ClpXP protease degrades the cytoskeletal protein, FtsZ, and modulates FtsZ polymer dynamics. AB - FtsZ is the major cytoskeletal protein in bacteria and a tubulin homologue. It polymerizes and forms a ring where constriction occurs to divide the cell. We found that FtsZ is degraded by E. coli ClpXP, an ATP-dependent protease. In vitro, ClpXP degrades both FtsZ protomers and polymers; however, polymerized FtsZ is degraded more rapidly than the monomer. Deletion analysis shows that the N terminal domain of ClpX is important for polymer recognition and that the FtsZ C terminus contains a ClpX recognition signal. In vivo, FtsZ is turned over slower in a clpX deletion mutant compared with a WT strain. Overexpression of ClpXP results in increased FtsZ degradation and filamentation of cells. These results suggest that ClpXP may participate in cell division by modulating the equilibrium between free and polymeric FtsZ via degradation of FtsZ filaments and protomers. PMID- 19541656 TI - Task-dependent organization of brain regions active during rest. AB - The human brain demonstrates complex yet systematic patterns of neural activity at rest. We examined whether functional connectivity among those brain regions typically active during rest depends on ongoing and recent task demands and individual differences. We probed the temporal coordination among these regions during periods of language comprehension and during the rest periods that followed comprehension. Our findings show that the topography of this "rest network" varies with exogenous processing demands. The network encompassed more highly interconnected regions during rest than during listening, but also when listening to unsurprising vs. surprising information. Furthermore, connectivity patterns during rest varied as a function of recent listening experience. Individual variability in connectivity strength was associated with cognitive function: more attentive comprehenders demonstrated weaker connectivity during language comprehension, and a greater differentiation between connectivity during comprehension and rest. The regions we examined have generally been thought to form an invariant physiological and functional network whose activity reflects spontaneous cognitive processes. Our findings suggest that their function extends beyond the mediation of unconstrained thought, and that they play an important role in higher-level cognitive function. PMID- 19541657 TI - Lateral inferior prefrontal cortex and anterior cingulate cortex are engaged at different stages in the solution of insight problems. AB - Two studies used puzzles that required participants to find a word that satisfied a set of constraints. The first study used a remote-association task, where participants had to find a word that would form compound words with 3 other words. The second study required participants to complete a word fragment with an associate of another word. Both studies produced distinct patterns of activity in the lateral inferior prefrontal cortex (LIPFC) and the anterior cingulate cortex (ACC). Activation in the LIPFC rose only as long as the participants were trying to retrieve the solution and dropped off as soon as the solution was obtained. However, activation in the ACC increased upon the retrieval of a solution, reflecting the need to process that solution. The data of the second experiment are fit by an information-processing model that interprets the activity in the LIPFC as reflecting retrieval operations and the activity in the ACC as reflecting subgoal setting. PMID- 19541658 TI - Dendro-dendritic bundling and shared synapses between gonadotropin-releasing hormone neurons. AB - The pulsatile release of gonadotropin-releasing hormone (GnRH) is critical for mammalian fertility, but the mechanisms underlying the synchronization of GnRH neurons are unknown. In the present study, the full extent of the GnRH neuron dendritic tree was visualized by patching and filling individual GnRH neurons with biocytin in acute brain slices from adult GnRH-green fluorescent protein (GFP) transgenic mice. Confocal analysis of 42 filled GnRH neurons from male and female adult mice revealed that the dendrites of the great majority of GnRH neurons (86%) formed multiple close appositions with dendrites of other GnRH neurons. Two types of interactions were encountered; the predominant interaction was one of vertical dendritic bundling where dendrites were found to wrap around each other in the same axis. The other interaction was one in which a GnRH neuron dendrite intercepted other GnRH neuron dendrites in a perpendicular fashion. Electron microscopy using pre-embedded, silver-enhanced immunogold labeling for both GnRH and GFP peptides in GnRH-GFP transgenic mice, confirmed that GnRH neuron dendrites were often immediately juxtaposed. Membrane specializations, including punctae and zonula adherens, were found connecting adjacent dendritic elements of GnRH neurons. Remarkably, individual afferent axon terminals were found to synapse with multiple GnRH neuron dendrites at sites of bundling. Together, these data demonstrate that GnRH neurons are not isolated from one another but, rather, interconnected via their long dendritic extensions. The observation of shared synaptic input to bundled GnRH neuron dendrites suggests a mechanism of GnRH neuron synchronization. PMID- 19541659 TI - Modulation of intein activity by its neighboring extein substrates. AB - Inteins comprise a large family of phylogenetically widespread self-splicing protein catalysts that colonize diverse host proteins. The evolutionary and functional relationship between the intein and the split-host protein, the exteins, is largely unknown. To probe an association, we developed an in vivo and in vitro intein assay based on FRET. The FRET assay reports cleavage of the intein from its N-terminal extein. Applying this assay to randomized extein libraries, we show that the nature of the extein substrate bordering the intein can profoundly influence intein activity. Residues proximal to the intein splicing junction in both N- and C-terminal exteins can accelerate the N-terminal cleavage rate by >4-fold or attenuate cleavage by 1,000-fold, both resulting in compromised self-splicing efficiency. The existence and the magnitude of extein effects require consideration for maximizing the utility of inteins in biotechnological applications, and they predict biases in intein integration sites in nature. PMID- 19541660 TI - Algorithmic design of self-assembling structures. AB - We study inverse statistical mechanics: how can one design a potential function so as to produce a specified ground state? In this article, we show that unexpectedly simple potential functions suffice for certain symmetrical configurations, and we apply techniques from coding and information theory to provide mathematical proof that the ground state has been achieved. These potential functions are required to be decreasing and convex, which rules out the use of potential wells. Furthermore, we give an algorithm for constructing a potential function with a desired ground state. PMID- 19541661 TI - A temporal analysis of antioxidative defense responses in the phloem of Picea abies after attack by Ips typographus. AB - The temporal gradation of antioxidants was investigated on the phloem tissue of Norway spruce [Picea abies (L.) Karst.] in response to weather conditions and colonization levels of Ips typographus L. (Col., Scolytidae). Two weeks after pheromone dispensers were placed on trees, the initial reaction of Norway spruce to bark beetle attack resulted in moderately lowered levels of total glutathione (tGSH) and total cysteine. Likewise, the total ascorbic acid dropped slightly below the control levels, whereas the concentration of dehydroascorbic acid increased in comparison to the first sampling date. This transient degradation and oxidation of the glutathione and ascorbate system was accompanied by moderately increased concentrations of total phenolics. One month later, the shift in antioxidant balance after moderate attack differed quantitatively from the reaction after massive attack. An intensification of antioxidant defense occurred within moderately affected bark. Total cysteine and tGSH contents were markedly raised, whereas the concentrations of total ascorbic acid and total phenolics were slightly increased by moderate attack. On the other hand, massive bark beetle colonization caused a strong decrease in tGSH and total phenolics, whereas total cysteine and total ascorbic acid values remained at control level. Dependent upon the intensity and the success of the attack, a progressive degradation of antioxidants was determined at later sampling dates, which was accompanied by an obvious oxidation of the ascorbate and glutathione pools. With an unsuccessful defense upon massive attack, the thiols and total phenolics did not reach a new steady state, but deteriorated until the end of the brood beetles' development. In contrast, the dynamic antioxidative response within the moderately affected trees indicated an acclimation stage in the middle of July. It was characterized by a higher accumulation of tGSH, total ascorbic acid and total phenolics as well as a more reduced redox state of glutathione. A sequence of changes in the endogenous levels of antioxidant defense molecules in the bark beetle-affected Norway spruce showed consistency with the general ecophysiological stress-response concept, and provided important avenues for evaluating the role and effectiveness of antioxidants in systemic acquired resistance against the complex interactive effects of bark beetle attack and environmental factors. PMID- 19541662 TI - Assessment of cardiovascular risk in paediatric peritoneal dialysis patients: a Turkish Pediatric Peritoneal Dialysis Study Group (TUPEPD) report. AB - METHODS: We aimed to clarify arteriosclerotic risk and to document possible relationships between cardiovascular risk factors and echocardiographic parameters in paediatric peritoneal dialysis (PD) patients. M-mode/Doppler/tissue Doppler echocardiographic studies and lipid/lipoproteins, homocysteine, high sensitivity C-reactive protein (HS-CRP) levels and carotid intima-media thickness (CIMT) were determined in 59 patients (age: 14.2 +/- 4.5 years) and in 36 healthy subjects. RESULTS: Structural and functional cardiac abnormalities were observed in patients on maintenance dialysis. Increased left ventricular mass index (LVMI, P = 0.000), relative wall thickness (P = 0.000), myocardial performance index (MPI, P = 0.000) were documented in the patients. Lipoprotein (a) (P = 0.000), homocysteine (P = 0.001), HS-CRP (P = 0.000) and CIMT (P = 0.000) were significantly elevated in the patients. Left ventricular hypertrophy (LVH) was prevalent in 68% of the patients. Patients with LVH had higher levels of HS-CRP (P = 0.001) and CIMT (P = 0.028) than those without LVH. Haemoglobin was an independent predictor of LVMI (beta: -8.9, P = 0.001), while residual diuresis and CIMT were independent predictors of diastolic dysfunction (beta: -0.45, P = 0.034 and beta: 5.90, P = 0.008, respectively). Albumin (beta: -0.72, P = 0.018) and Kt/V urea (beta: -0.48, P = 0.012) were significant predictors of CIMT. There were positive correlations between LVMI and CIMT. HS-CRP was positively correlated with LVMI as well as CIMT. CONCLUSIONS: Elevated levels of atherosclerotic/ inflammatory risk factors, low haemoglobin levels and loss of residual renal function and their negative effects on heart are of remarkable importance in paediatric patients on maintenance peritoneal dialysis. Achieving recommended targets for haemoglobin, blood pressure and Kt/V urea, preserving residual renal function as well as managing inflammation and subsequent arteriosclerosis is obviously essential to improve the patients' prognosis. PMID- 19541663 TI - Nail-patella syndrome, infantile nephrotic syndrome: complete remission with antiproteinuric treatment. PMID- 19541666 TI - The Jupiter trial--new territory for statins? PMID- 19541664 TI - Total cholesterol content of erythrocyte membranes and coronary atherosclerosis: an intravascular ultrasound pilot study. AB - BACKGROUND: Increasing evidence suggests that erythrocytes may participate in atherogenesis. We sought to investigate the relationship between total cholesterol content in erythrocyte membranes (CEM) and coronary atheroma burden in patients with coronary artery disease (CAD). METHODS: We prospectively enrolled 28 participants: 11 patients with angiographically significant CAD and 17 controls. Intravascular ultrasound (IVUS) and 3-dimensional reconstruction of coronary arteries was performed in the patient subgroup. RESULTS: Cholesterol content of erythrocyte membranes was higher in patients compared to controls (P < .01). Cholesterol content of erythrocyte membranes correlated with total atheroma volume (r = .82, P < .01) and with percentage plaque area at the vessel site with minimal lumen area (r = .75, P < .05). On multivariate analysis, CEM was the only variable independently predicting total atheroma volume (P = .05). CONCLUSIONS: This pilot study is the first to demonstrate a significant relation between CEM and coronary atherosclerotic burden, suggesting a possible role of erythrocyte membrane-derived lipids in the expansion of atheromata. The results merit validation in larger studies. PMID- 19541667 TI - Tissue Doppler imaging must be performed to detect early left ventricular dysfunction in patients with type 1 diabetes mellitus. AB - AIMS: We sought to compare diastolic parameters in patients having type 1 diabetes mellitus (DM) and healthy controls using both pulse-wave (PW) Doppler and relatively novel tissue-Doppler imaging (TDI) to evaluate the possible effect of diabetes on left ventricular dysfunction. METHODS AND RESULTS: One hundred and thirty-two patients were evaluated (81 type 1 diabetic patients and 51 healthy volunteers). The detailed M-mode, two-dimensional, colour Doppler; PW Doppler; and TDI analyses were performed on resting subjects in a regular setting. Posterior wall thickness, left atrial indexed diameter, and A velocity were significantly higher in the diabetics when compared with control group (P = 0.019, <0.001, 0.033, respectively). Rest of the M-mode and PW Doppler parameters of diabetics were comparable with those of control subjects (P > 0.05 for all). However, both septal E' and lateral E' velocities were significantly lower in diabetics than in the control subjects on TDI echocardiographic examination (P < 0.001 and 0.011, respectively). In addition, E'/septal E' and E/lateral E' ratios were significantly higher in the diabetic group (P < 0.001 and 0.008, respectively). CONCLUSION: TDI is a more accurate and powerful method than PW or M-mode in determination of early cardiac involvement related to type 1 DM even in the subclinical phase as well as hereditary cardiomyopathies. PMID- 19541668 TI - ERalpha, ERbeta, and gpER: novel aspects of oestrogen receptor signalling in atherosclerosis. PMID- 19541670 TI - The utilization of ultrasound and microbubbles for therapy in acute coronary syndromes. AB - Ultrasound has become a useful high resolution imaging modality for examining the cardiac microcirculation. With the use of microbubbles as an ultrasound contrast agent, ultrasound can be utilized to image the microcirculation and detect capillary flow abnormalities in acute ischaemia. A wide range of ultrasound frequencies (including those used for diagnostic transthoracic imaging) have also been utilized therapeutically to augment the effectiveness of fibrinolytic therapy in ST-segment elevation myocardial infarction (STEMI). Ultrasound and microbubbles are now being explored as methods of improving both microcirculatory and epicardial flow in acute STEMI. This article will review the mechanisms by which ultrasound and microbubbles assist in thrombus detection and dissolution. In addition, the pre-clinical studies utilizing transthoracic ultrasound as a therapeutic entity in acute STEMI will be reviewed. Clinical studies, completed and ongoing, will also be presented. PMID- 19541669 TI - Endothelium-specific overexpression of human IC53 downregulates endothelial nitric oxide synthase activity and elevates systolic blood pressure in mice. AB - AIMS: Hypertension is one of the major risk factors for cardiovascular diseases. Endothelial cells (ECs) exert important functions in the regulation of blood pressure. A novel gene, IC53, as an isoform of the cyclin-dependent kinase (CDK) binding protein gene C53, is mainly expressed in vascular ECs and is upregulated in the failing heart of rats. Overexpression of IC53 promotes proliferation of ECs. To examine whether IC53 plays a role in the regulation of vascular tone and blood pressure, we constructed a transgenic (tg) mouse model of the IC53 gene and studied its phenotypes relevant to vascular function. METHODS AND RESULTS: IC53 cDNA was cloned from a human aorta cDNA library. Using the endothelium-specific VE-cadherin promoter, we constructed tg mice in which IC53 was specifically overexpressed in vascular endothelia and found that the tg mice exhibit elevated systolic blood pressure (SBP) in contrast to the wild-type (wt) controls. Further studies revealed impaired endothelium-dependent vasodilation, reduced nitric oxide (NO) production and decreased endothelial NO synthase (eNOS) expression, and activity in the tg mice. Inhibition of IC53 in human umbilical vein ECs induces upregulation of eNOS activity. CONCLUSION: Our results indicate that IC53 participates in the regulation of vascular homeostasis. Endothelium-specific overexpression of IC53 is associated with elevated SBP, which may be in part attributed to the downregulation of eNOS signalling. PMID- 19541671 TI - Serotonin enhances platelet procoagulant properties and their activation induced during platelet tissue factor uptake. AB - AIMS: Circulating tissue factor (TF) has been linked to thrombus propagation. Our group demonstrated that platelets possess mechanisms to capture TF-rich microvesicles (TF-MVs). Serotonin facilitates the development of platelets with increased procoagulant activity. An enhanced platelet serotonin uptake has been identified with increased cardiovascular risk. We have investigated the involvement of serotonergic mechanisms facilitating the interaction of human platelets with TF-MVs. Inhibitory strategies aimed at blocking serotonin and coagulation mechanisms were also studied. METHODS AND RESULTS: Standard aggregometry, flow cytometry, electron microscopy, and thrombin generation assays were performed. TF-MVs induced platelet aggregation in heparinized platelet-rich plasma (PRP) samples; this aggregation was further accelerated by serotonin. In washed platelets, serotonin enhanced platelet aggregation to TF-MVs with a maximum peak of 55.9 +/- 1.8 vs. 48.7 +/- 2.1% (P < 0.05). Inhibitory strategies with a selective serotonin re-uptake inhibitor and with lepirudin decreased these aggregations. Ultrastructural analysis revealed that serotonin induced platelet pseudopodia formation, thus facilitating the engulfment of TF-MVs. In general, serotonin significantly enhanced (P < 0.05) thrombin generation and the expression of activation markers and procoagulant activity in platelets measured for TF-MVs alone. CONCLUSION: Serotonin enhances the interaction of platelets with TF-MVs, increases platelet activation, and potentiates their overall procoagulant activity. The present results could have significant implications in thrombus formation and in the thrombogenic profile of pathological situations with increased cardiovascular risk. PMID- 19541673 TI - Development of generic calcium imaging assay for monitoring Gi-coupled receptors and G-protein interaction. AB - G-protein-coupled receptors (GPCRs) are important therapeutic targets for many areas of drug research and development. Although chimeric Galpha16 proteins are valuable tools for detecting the activation of Galpha(i/o)-coupled receptors, the details of the activation process remain unclear. The authors introduce a series of chimeras that combine both Galpha16 and Galpha(i/o) (Galpha(16/o), Galpha(16/i2), and Galpha(16/i3)) into a well-established transient expression system to examine the ability of these chimeras to interact with D2 long-form (D2L) dopamine and 5-HT1A serotonin receptors. The pEC50 data obtained for known agonists were similar to results from previous studies that used other cell-based assays, thus indicating sufficient sensitivity for the assay. Moreover, quinpirole exhibited similar intrinsic activity to dopamine at the D2L receptor, whereas S-(-)-3-PPP displayed partial activity of dopamine and quinpirole in the presence of the Galpha(16/o) chimera. The potency of dopamine for D2L receptors was similar among Galpha(16/o), Galpha(16/i2), and Galpha(16/i3). In contrast, the 5-HT1A receptor exhibited a significantly preferential coupling for Galpha(16/i3) compared with Galpha(16/i2) when serotonin was used as a ligand. This finding was in close agreement with the results of previous reports. The present system could therefore be used as a rapid functional assay for high throughput screening and deorphanization. PMID- 19541672 TI - Diabetes-related support, regimen adherence, and health decline among older adults. AB - OBJECTIVES. Social support is generally conceptualized as health promoting; however, there is little consensus regarding the mechanisms through which support is protective. Illness support has been proposed to promote regimen adherence and subsequent prevention of health decline. We hypothesize that (a) support for regimen adherence is negatively associated with self-reported health decline among older diabetic adults and that (b) regimen adherence is negatively associated with health decline among older diabetic adults. METHODS: We used the Health and Retirement Study data on individuals over the age of 60 years with type 2 diabetes mellitus (n = 1,788), examining change in self-reported health status over a 2-year period using binomial and cumulative ordinal logistic regression models. RESULTS: Diabetic support is not significantly associated with health decline, but it is strongly associated with adherence to health-promoting activities consisting of a diabetic regimen. Therefore, the extent to which one receives illness support for a given regimen component is highly positively associated with adhering to that component, although this adherence does not necessarily translate into protection against perceived decline in health. CONCLUSIONS: Illness-related support appears to be a mechanism through which social support matters in the diabetic population. Although this relationship did not extend to prevention of health status decline among diabetics, the relationship between support and illness management is promising. PMID- 19541674 TI - Workshop on ethical issues and justification for high-dose budget-limited procedures: a report from an exploratory and consensus meeting held at Hairmyres Hospital, Scotland. AB - This summary report presents aspects of work that was carried out as part of the European Commission SENTINEL Project (Safety and Efficacy for New Techniques and Imaging using New Equipment to Support European Legislation). It arises from the discussions at a workshop organised by the first author at Hairmyres Hospital, East Kilbride in September 2006. The second author facilitated the workshop and moderated the discussion. The paper notes key points from the discussion and provides some pertinent comments. PMID- 19541675 TI - Protecting the brain from gaseous and solid micro-emboli during coronary artery bypass grafting: a randomized controlled trial. AB - AIMS: The purpose of the study was to investigate whether intra-operative filter devices protect the brain during coronary artery bypass grafting (CABG) and to determine the impact of solid and gaseous micro-emboli on neuropsychological functioning. METHODS AND RESULTS: Patients undergoing CABG received either an intra-aortic filter (Embol-X) (n = 43), designed to reduce solid micro-emboli, a dynamic bubble trap (DBT) (n = 50), designed to reduce gaseous micro-emboli, or no additional device (control group) (n = 50). Cognitive functioning was assessed before and 3 months after CABG. Micro-emboli signals (MES) were detected during surgery using transcranial Doppler (TCD) sonography. Cerebral magnetic resonance imaging (MRI) was carried out before and after surgery. Primary endpoint was the cognitive outcome of the filter groups compared with the controls. Analysis of covariance was performed using the post-operative cognitive test scores as continuous variables in covariance of the corresponding pre-operative scores. Secondary endpoints were the MES rates and the number of acute ischaemic lesions after CABG. Compared with the controls, cognitive functioning of the DBT group was better in executive functioning (t = 2.525, P = 0.0065) and verbal short-term memory (t = 2.420, P = 0.009). The Embol-X group did not perform better in any test. The total number of MES was lower in the DBT group (median 99, P = 0.0019), but not in the Embol-X group (median 162.5, P > 0.05), both compared with controls (median 164.5). After surgery, 17 patients displayed small ischaemic brain lesions on MRI with equal distribution between the groups. CONCLUSION: Gaseous micro-embolization contributes to neuropsychological decline, which is measurable 3 months post-operatively. No filter device could protect the brain during CABG completely. However, the use of the DBT tends to improve the cognitive outcome after CABG. Gas filters are recommendable for neuroprotection during cardiac surgery. PMID- 19541677 TI - Predicting the efficacy of convection warming in anaesthetized children. AB - BACKGROUND: We previously described a convection warming technique (Cassey J, Armstrong P, Smith GE, Farrell PT. Paediatr Anaesth 2006; 16: 654-62). This study further analyses the children in that original study with three aims: (i) to investigate factors purported to influence children's heating rates, (ii) to describe the most effective usage of this warming technique, and (iii) to understand better the physiology of convection warming. METHODS: Children having anaesthesia for elective surgery lasting longer than 90 min in ambient temperature 21 degrees C were warmed by a 'Bair Hugger' attached to a custom built heat dissipation unit. Relationships between child and procedure characteristics and various thermal measures were analysed, and a thermodynamic model was evaluated. RESULTS: Thirty-nine children (aged 2 days to 12.5 yr) were studied. There were statistically significant correlations between a number of factors (e.g. height and weight) and heating efficacy. Our model demonstrated the impact of changing patient characteristics on temperature profiles. Neither the morphological characteristics nor our model could predict an individual's T(core) behaviour. CONCLUSIONS: (i) Although the effectiveness of this warming technique is influenced by patient/procedure characteristics, these do not predict normothermia (uncertainty +/-28 min). Effectiveness is independent of simple thermal measures. (ii) Previously described measures of vasoconstriction are not valid in children. (iii) Our model shows children's thermal properties change with their T(core). However, key factors are unknown for an individual and our model does not predict heating efficacy. (iv) To minimize the risk of hyperthermia, we recommend continuous measurement of T(core) during convection heating. The device air temperature should be turned to medium (38 degrees C) as T(core) approaches 37 degrees C. PMID- 19541676 TI - The impact of ataxin-1-like histidine insertions on polyglutamine aggregation. AB - Spinocerebellar ataxia type 1 (SCA1) is one of a group of nine expanded CAG repeat diseases, in which polyglutamine (polyQ) expansion above a threshold is associated with increased disease risk and aggregation. SCA1 is unique in which the polyQ in the disease protein, ataxin1, often contains a few His residues that appear to block toxicity. Here, we ask how His insertions affect aggregation by comparing a Q(30) peptide with and without a centrally inserted His-Gln-His sequence. We found that at pH 7.5-8.5, His interruptions decrease polyQ aggregation rates but do not change the spontaneous growth mechanism: nucleated growth polymerization with a critical nucleus of one without non-fibrillar intermediates. The decreased aggregation rates are because of reductions in nucleation equilibrium constants. At pH 6, however, the His-interrupted peptide aggregates by a different mechanism that involves a low ThT-binding intermediate and produces a polymorphic amyloid product. In aggregates grown at pH 7.5, the His residues are solvent-accessible. Aggregates of His-inserted polyQ are good seeds for Q(30) elongation, suggesting the potential to recruit polyQ proteins in the cell. Our data are therefore most consistent with His insertions blocking toxicity by suppressing rates and/or altering pathways of spontaneous aggregation. PMID- 19541678 TI - Indices of pulmonary oxygenation in pathological lung states: an investigation using high-fidelity, computational modelling. AB - BACKGROUND: Existing indices of pulmonary oxygenation vary misleadingly with external factors such as inspired oxygen fraction (FI(O2)), arterial carbon dioxide tension (PaCO2), and haemoglobin (Hb). Previous work suggested that some indices may be acceptably useful in particular scenarios such as acute respiratory distress syndrome (ARDS) or where FI(O2)>60%. However, it is not possible to identify such scenarios in most clinical contexts; therefore we aimed to examine the induced variability of existing indices in a population of patients with a variety of lung defects. METHODS: We configured nine virtual patients within the Nottingham Physiology Simulator, each with a unique pulmonary configuration but identical arterial blood gases at FI(O2) 30%, PaCO2 6.0 kPa and Hb 8.0 g dl(-1). Factors (FI(O2), P(CO2), Hb) were varied independently and indices of oxygenation including calculated venous admixture (Qs/Qt), arterial oxygen tension (PaO2/FI(O2)), arterio-alveolar gas tension gradient (PA-aO2), and respiratory index (PA-aO2/PaO2) were recorded. RESULTS: All indices varied with FI(O2), with greatest variation with lung defects having least true (absolute) shunt. Calculated Qs/Qt resisted induced variation best of all the indices, but varied by 30% of its mean value during FI(O2) variation. PaO2/FI(O2) varied greatly, especially during variation in FI(O2) (up to 74% of its average value), and most markedly in defects with little true (absolute) shunt. PaCO2 and Hb variation caused small, consistent changes in all indices that were similar between lung-states. CONCLUSIONS: No existing index of oxygenation adequately describes the severity of gas exchange defect. Existing indices of oxygenation vary with disease severity, disease type, and external factors such as FI(O2). A novel and robust index is needed. PMID- 19541679 TI - Addition of clonidine or dexmedetomidine to bupivacaine prolongs caudal analgesia in children. AB - BACKGROUND: Caudal block is a common technique for paediatric analgesia but with the disadvantage of short duration of action after single injection. Caudal dexmedetomidine and clonidine could offer significant analgesic benefits. We compared the analgesic effects and side-effects of dexmedetomidine and clonidine added to bupivacaine in paediatric patients undergoing lower abdominal surgeries. METHODS: Sixty patients (6 months to 6 yr) were evenly and randomly assigned into three groups in a double-blinded manner. After sevoflurane in oxygen anaesthesia, each patient received a single caudal dose of bupivacaine 0.25% (1 ml kg(-1)) combined with either dexmedetomidine 2 microg kg(-1) in normal saline 1 ml, clonidine 2 microg kg(-1) in normal saline 1 ml, or corresponding volume of normal saline according to group assignment. Haemodynamic variables, end-tidal sevoflurane, and emergence time were monitored. Postoperative analgesia, use of analgesics, and side-effects were assessed during the first 24 h. RESULTS: Addition of dexmedetomidine or clonidine to caudal bupivacaine significantly promoted analgesia time [median (95% confidence interval, CI): 16 (14-18) and 12 (3-21) h, respectively] than the use of bupivacaine alone [median (95% CI): 5 (4 6) h] with P<0.001. However, there was no statistically significant difference between dexmedetomidine and clonidine as regards the analgesia time (P=0.796). No significant difference was observed in incidence of haemodynamic changes or side effects. CONCLUSIONS: Addition of dexmedetomidine or clonidine to caudal bupivacaine significantly promoted analgesia in children undergoing lower abdominal surgeries with no significant advantage of dexmedetomidine over clonidine and without an increase in incidence of side-effects. PMID- 19541680 TI - Not so innocent bystander(s). AB - A patient with atrial flutter, intermittent non-sustained wide complex tachycardia and 1:1 AV conduction is reported. Electrophysiology testing showed counterclockwise isthmus-dependent right atrial flutter with conduction via the AV node and an innocent bystander left lateral accessory pathway. This explained the observed intermittent wide complex tachycardia. After successful bidirectional cavotricuspid isthmus conduction block, a sustained wide complex tachycardia with identical counterclockwise right atrial activation and rate occurred. This was due to antidromic AV re-entrant tachycardia with innocent bystander activation of the right atrium mimicking atrial flutter. Accessory pathway ablation effectively stopped tachycardia. PMID- 19541681 TI - Image integration increases efficacy of paroxysmal atrial fibrillation catheter ablation: results from the CartoMerge Italian Registry. AB - AIMS: The aim of this study was to investigate whether circumferential pulmonary vein (PV) isolation guided by image integration improves the procedural and clinical outcomes of atrial fibrillation (AF) ablation in comparison with segmental PV isolation and circumferential PV isolation guided by three dimensional (3D) electroanatomical mapping alone. METHODS AND RESULTS: Procedural and clinical outcomes of 573 patients who underwent their first catheter ablation for paroxysmal AF between January 2005 and April 2007 were collected from 12 centres. We evaluated three techniques: segmental ostial PV isolation (SOCA group, 240 patients), circumferential PV isolation guided by electroanatomical mapping (CARTO group, 107 patients), and circumferential PV isolation guided by electroanatomical mapping integrated with magnetic resonance/computed tomographic images of the left atrium (MERGE group, 226 patients). Procedure duration proved to be shorter in MERGE group patients than in CARTO group patients (P < 0.04), but longer than in SOCA group patients (P < 0.0001). During follow-up, atrial tachyarrhythmias relapsed more frequently in SOCA group patients (44.6%) and CARTO group patients (41.7%) than in MERGE group patients (22.6%; P < 0.0001). CONCLUSION: In patients with paroxysmal AF, circumferential PV isolation guided by image integration significantly improves clinical outcome in comparison with both circumferential PV isolation guided by 3D mapping alone and with segmental electrophysiologically guided PV isolation. PMID- 19541682 TI - Risk factors for respiratory syncytial virus hospitalisation in children with heart disease. AB - OBJECTIVE: To assess the risk and risk factors for respiratory syncytial virus (RSV) hospitalisation and determinants of the severity of RSV disease in children with heart disease. METHODS: By using a database on RSV tests in Denmark all children with RSV diagnosed with heart disease in Denmark from January 1996 to April 2003 were identified. For each case child one control child matched for age and centre was drawn from the population of children with heart disease. Clinical information was obtained through a review of all records. RESULTS: Data were obtained on 313 pairs. Median age at admission was 280 days (range 15-2379). In the multivariate analysis predictors of RSV hospitalisation were Down syndrome (odds ratio (OR) 3.24, 95% CI 1.80 to 5.80), cardiomyopathy (OR 5.84, 95% CI 1.26 to 27.16) and haemodynamically significant heart disease (OR 1.53, 95% CI 1.04 to 2.26). During RSV hospitalisation predictors of the need for respiratory support (supplemental oxygen, nasal continuous positive airway pressure or mechanical ventilation) were young age (relative risk (RR) 0.47, 95% CI 0.32 to 0.67 per additional year of age) and cardiac decompensation (RR 1.81, 95% CI 1.02 to 3.23). The incidence rate of RSV hospitalisation among children with any heart disease aged 0-23 months was 5.65 per 100 child-years. CONCLUSION: In children with heart disease risk factors for RSV admission are Down syndrome, cardiomyopathy and haemodynamically significant heart disease. Young age and cardiac decompensation are associated with a more severe course of RSV disease. PMID- 19541683 TI - Molecular profile of ductal carcinoma in situ of the breast in BRCA1 and BRCA2 germline mutation carriers. AB - AIMS: Ductal carcinoma in situ (DCIS) is an established late precursor of sporadic invasive breast cancer and to a large extent parallels its invasive counterpart with respect to molecular changes and immunophenotype. Invasive breast cancers in germline BRCA1 and BRCA2 mutation carriers have a distinct "basal" and "luminal" immunophenotype, respectively, but the immunophenotype of their precursor lesions has hardly been studied, and this was the aim of this study. METHODS: DCIS lesions of 25 proven BRCA1 and 9 proven BRCA2 germline mutation carriers and their 22 and 6, respectively, accompanying invasive lesions were stained by immunohistochemistry for oestrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor (HER)2/neu, cytokeratin (CK)5/6, CK14, epidermal growth factor receptor (EGFR) and Ki67. RESULTS: DCIS lesions in BRCA1 mutation carriers were mostly of the basal molecular type with low ER/PR/HER2 expression, while they frequently expressed CK5/6, CK14 and EGFR, and were mostly grade 3 and highly proliferative. DCIS lesions in BRCA2 mutation carriers were mostly of luminal molecular type with frequent expression of ER/PR, and infrequent expression of CK5/6, CK14 and EGFR, and they were mostly grade 3 and showed low proliferation. In BRCA1 and BRCA2 mutation carriers there was a high concordance between DCIS lesions and their concomitant invasive counterpart with regard to expression of individual markers as well as "molecular" subtype. CONCLUSIONS: Although the number of cases studied was low, DCIS lesions in BRCA1 and BRCA2 mutations carriers are usually of the basal and luminal molecular type, respectively, similar to their accompanying invasive cancers, thereby providing evidence that DCIS is a direct precursor lesion in these hereditary predisposed patients. This also suggests that crucial carcinogenetic events leading to these phenotypes in hereditary predisposed patients occur before the stage of invasion. PMID- 19541684 TI - Trisomy 7 and 17 mark papillary renal cell tumours irrespectively of variation of the phenotype. AB - BACKGROUND: Papillary renal cell tumours (RCTs) have been described as a genetic entity. Recently, papillary RCTs have been divided into small (type 1) and large (type 2) cell tumours. Subsequent DNA analyses have resulted in controversial data regarding putative genetic changes marking type 1 and type 2 tumours. AIM: The aim of this study was to improve the original description that papillary RCT is a genetic entity regardless of the phenotypic variation. METHODS: DNA from 163 papillary RCTs, including 82 multiplex tumours from eight hereditary cases, was analysed for copy number changes by chromosomal comparative genomic hybridisation (CGH) and/or for allelic changes at chromosomes 7 and 17 by microsatellite analysis. The results of the genetic analysis were compared with the cytological characteristics of the tumours. RESULTS: The results showed alterations of chromosomes 7 and 17 at similar frequencies in papillary RCTs with characteristics ranging from small to large cell, nuclear grade 1 to 3, and 3 mm to 16 cm diameter. CONCLUSION: Trisomies of chromosomes 7 and 17 are specific genetic alterations in papillary RCTs irrespective of their size, grade and cellular differentiation. PMID- 19541685 TI - Mutation prediction models in Lynch syndrome: evaluation in a clinical genetic setting. AB - BACKGROUND/AIMS: The identification of Lynch syndrome is hampered by the absence of specific diagnostic features and underutilisation of genetic testing. Prediction models have therefore been developed, but they have not been validated for a clinical genetic setting. The aim of the present study was to evaluate the usefulness of currently available prediction models. METHODS: The authors collected data of 321 index probands who were referred to the department of clinical genetics of the Erasmus Medical Center because of a family history of colorectal cancer. These data were used as input for five previously published models. External validity was assessed by discriminative ability (AUC: area under the receiver operating characteristic curve) and calibration. For further insight, predicted probabilities were categorised with cut-offs of 5%, 10%, 20% and 40%. Furthermore, costs of different testing strategies were related to the number of extra detected mutation carriers. RESULTS: Of the 321 index probands, 66 harboured a germline mutation. All models discriminated well between high risk and low risk index probands (AUC 0.82-0.84). Calibration was well for the Premm(1,2) and Edinburgh model, but poor for the other models. Cut-offs could be found for the prediction models where costs could be saved while missing only few mutations. CONCLUSIONS: The Edinburgh and Premm(1,2) model were the models with the best performance for an intermediate to high risk setting. These models may well be of use in clinical practice to select patients for further testing of mismatch repair gene mutations. PMID- 19541686 TI - The spectrum of structural abnormalities on CT scans from patients with CF with severe advanced lung disease. AB - RATIONALE: In cystic fibrosis (CF), lung disease is the predominant cause of morbidity and mortality. Little is known about the spectrum of structural abnormalities on CT scans from patients with CF with severe advanced lung disease (SALD). No specific CT scoring system for SALD is available. OBJECTIVES: To design a quantitative CT scoring system for SALD, to determine the spectrum of structural abnormalities in patients with SALD and to correlate the SALD system with an existing scoring system for mild CF lung disease and pulmonary function tests (PFTs). METHODS: 57 patients with CF contributed one CT made during screening for lung transplantation. For the SALD system, lung tissue was divided into four components: infection/inflammation (including bronchiectasis, airway wall thickening, mucus and consolidations), air trapping/hypoperfusion, bulla/cysts and normal/hyperperfused tissue. The volume proportion of the components was estimated on a 0-100% scale; mean volumes for the whole lung were computed. Scores were correlated with Brody-II scores and PFTs. RESULTS: The SALD system identified a wide spectrum of structural abnormalities ranging from predominantly infection/inflammation to predominantly air trapping/hypoperfusion. SALD infection/inflammation scores correlated with Brody-II scores (r(s) = 0.36 0.64) and SALD normal/hyperperfusion scores correlated with forced expiratory volume in 1 s (FEV(1); r(s) = 0.37). Reproducibility for both systems was good. CONCLUSIONS: A CT scoring system was developed to characterise the structural abnormalities in patients with SALD. A wide spectrum was observed in SALD, ranging from predominantly air trapping to predominantly infection/inflammation related changes. This spectrum may have clinical implications for patients with SALD. PMID- 19541687 TI - Microbleeds and post-stroke emotional lability. AB - OBJECTIVE: The clinical significance of microbleeds (MBs) in the development of psychiatric conditions following a stroke is unknown. Lesions located in various cortical and subcortical areas are thought to be involved in the pathophysiology of post-stroke emotional lability (PSEL). This study examined the association between PSEL and MBs. METHODS: A total of 519 Chinese patients with acute ischaemic stroke consecutively admitted to the acute stroke unit of a university affiliated regional hospital in Hong Kong were screened for PSEL 3 months after their index stroke. The number and location of MBs were evaluated with MRI. RESULTS: According to Kim's criteria, 74 (14.3%) patients had PSEL. In comparison with the non-PSEL group, patients in the PSEL group were more likely to have MBs in the thalamus as a whole (16.2% vs 6.5%; p = 0.004), its anterior (6.9% vs 2.0%, p = 0.02) and paramedian territories (8.1% vs 3.1%; p = 0.04), and a higher number of MBs in the entire brain (1.7+3.4 vs 1.3+5.0; p = 0.031). MBs in the thalamus remained an independent predictor of PSEL in the multivariate analysis, with an odds ratio of 4.7 (p = 0.002). CONCLUSION: Our results suggest that MBs in the thalamus may play a role in the development of PSEL. The importance of MBs in PSEL and other psychiatric conditions in stroke survivors warrants further investigation. PMID- 19541688 TI - Somatosensory temporal discrimination in patients with primary focal dystonia. AB - PURPOSES: To determine whether somatosensory temporal discrimination will reliably detect subclinical sensory impairment in patients with various forms of primary focal dystonia. METHODS: The somatosensory temporal discrimination threshold (STDT) was tested in 82 outpatients affected by cranial, cervical, laryngeal and hand dystonia. Results were compared with those for 61 healthy subjects and 26 patients with hemifacial spasm, a non-dystonic disorder. STDT was tested by delivering paired stimuli starting with an interstimulus interval of 0 ms followed by a progressively increasing interstimulus interval. RESULTS: STDT was abnormal in all the different forms of primary focal dystonias in all three body regions (eye, hand and neck), regardless of the distribution and severity of motor symptoms. Receiver operating characteristic curve analysis calculated in the three body regions yielded high diagnostic sensitivity and specificity for STDT abnormalities. CONCLUSIONS: These results provide definitive evidence that STDT abnormalities are a generalised feature of patients with primary focal dystonias and are a valid tool for screening subclinical sensory abnormalities. PMID- 19541689 TI - White matter hyperintensities and medial temporal lobe atrophy in clinical subtypes of mild cognitive impairment: the DESCRIPA study. AB - BACKGROUND: Clinical subtypes of mild cognitive impairment (MCI) may represent different underlying aetiologies. METHODS: This European, multicentre, memory clinic based study (DESCRIPA) of non-demented subjects investigated whether MCI subtypes have different brain correlates on MRI and whether the relation between subtypes and brain pathology is modified by age. Using visual rating scales, medial temporal lobe atrophy (MTA) (0-4) and white matter hyperintensities (WMH) (0-30) were assessed. RESULTS: Severity of MTA differed between MCI subtypes (p<0.001), increasing from a mean of 0.8 (SD 0.7) in subjective complaints (n = 77) to 1.3 (0.8) in non-amnestic MCI (n = 93), and from 1.4 (0.9) in single domain amnestic MCI (n = 70) to 1.7 (0.9) in multiple domain amnestic MCI (n = 89). The association between MCI subtype and MTA was modified by age and mainly present in subjects >70 years of age. Severity of WMH did not differ between MCI subtypes (p = 0.21). However, the combination of MTA and WMH differed between MCI subtypes (p = 0.02) CONCLUSION: We conclude that MCI subtypes may have different brain substrates, especially in older subjects. Isolated MTA was mainly associated with amnestic MCI subtypes, suggesting AD as the underlying cause. In non-amnestic MCI, the relatively higher prevalence of MTA in combination with WMH may suggest a different pathophysiological origin. PMID- 19541690 TI - Long-term effects of the Niigata-Chuetsu earthquake in Japan on acute myocardial infarction mortality: an analysis of death certificate data. AB - OBJECTIVE: To determine if the Niigata-Chuetsu earthquake of October 2004 increased long-term mortality from acute myocardial infarction (AMI). DESIGN: A comparative study of mortality rates before and after the earthquake, as well as between the disaster and control areas, by analysing death certificate data from 1 October 1999 to 30 September 2007. SETTING: The disaster area and a control area in Niigata Prefecture (n = 2 448 025 in 1 October 2004) in Japan. POPULATION: The total population of Niigata Prefecture observed for five years (12 333 429 person-years) before and three years (7 279 076 person-years) after the earthquake. MAIN OUTCOME MEASURES: Mortality from AMI (ICD-10, I21 and I22). RESULTS: Overall mortality rates from AMI five years before and three years after the earthquake in the disaster area were 47.3 and 53.9 per 100 000 person-years, respectively. Change (+6.6 or +14.0%) was significantly different (p = 0.0008), compared to the control area, where mortality rates were 42.5 and 42.6 per 100 000 person-years, respectively, and was not significantly different (p = 0.9028). In men, a change in AMI mortality before and after the earthquake in the disaster area was +7.1 per 100 000 person-years (+13.4%, p = 0.0172), and +2.0 (+4.2%, p = 0.2362) in the control area. In women, a change in AMI mortality in the disaster area was +6.2 per 100 000 person-years (+14.9%, p = 0.0184) and -1.6 (-4.2%, p = 0.2735) in the control area. CONCLUSIONS: The Niigata-Chuetsu earthquake significantly increased long-term mortality from AMI in both men and women. Clinicians and policymakers in public health must recognise the need for long term prevention of AMI in earthquake disaster areas. PMID- 19541691 TI - Collagen-gentamicin implant for prevention of sternal wound infection; long-term follow-up of effectiveness. AB - In a previous randomized controlled trial (LOGIP trial) the addition of local collagen-gentamicin reduced the incidence of postoperative sternal wound infections (SWI) compared with intravenous prophylaxis only. Consequently, the technique with local gentamicin was introduced in clinical routine at the two participating centers. The aim of the present study was to re-evaluate the technique regarding the prophylactic effect against SWI and to detect potential shifts in causative microbiological agents over time. All patients in this prospective two-center study received prophylaxis with application of two collagen-gentamicin sponges between the sternal halves in addition to routine intravenous antibiotics. All patients were followed for 60 days postoperatively. From January 2007 to May 2008, 1359 patients were included. The 60-day incidences of any SWI was 3.7% and of deep SWI 1.5% (1.0% mediastinitis). Both superficial and deep SWI were significantly reduced compared with the previous control group (OR=0.34 for deep SWI, P<0.001). There was no increase in the absolute incidence of aminoglycoside resistant agents. The majority of SWI were caused by coagulase negative staphylococci (CoNS). The incidence of deep SWI caused by Staphylococcus aureus was 0.07%. The results indicate a maintained effect of the prophylaxis over time without absolute increase in aminoglycoside resistance. (ClinicalTrials.gov NCT00484055). PMID- 19541692 TI - Thoracoscopic removal of a transesophageal ingested mediastinal foreign body. AB - Transesophageal migration of a foreign body into the mediastinum in human adults is rare. This event can result in life-threatening situations and is associated with a high mortality rate. Only a few case reports of thoracoscopic removal of a foreign body in adults have been published so far. The authors describe the case of a successfully performed thoracoscopic approach to remove an ingested needle from the mediastinum posterior of a 69-year-old patient. The authors are discussing why, in similar cases, removal of the foreign body should be performed and they describe the preliminary conditions for this method. A video file is attached. PMID- 19541693 TI - Comprehensive analysis of 4205 patients with chest trauma: a 10-year experience. AB - Thoracic trauma is one of the leading causes of morbidity and mortality in developing countries. In this retrospective study, we present our 10-year experience in the management and clinical outcome of 4205 cases with chest trauma associated with blunt and penetrating injuries in a level I trauma hospital in Turkey. In 66% of the cases, blunt injury mostly related to traffic accidents was the cause of chest trauma. Additional organ injuries were found in 35% (n=1471). Conservative treatment was administered for most patients. Tube thoracostomy was administered in 40% of all cases, whereas thoracotomy was performed in 6% (n=252), of which 209 were early interventions (P=0.001). The morbidity rate in all victims was 25.2%. The mortality rate was 9.3% for all patients and was 6.8% in blunt, 1.4% in penetrating, and 17.7% in associated organ injuries. Mortality and injury severity score (ISS) increased in patients having early surgery (P=0.001). Although most patients could be managed with conservative approaches, early thoracotomy was required in some cases. We believe that urgent hospital admission, early diagnosis, and multidisciplinary approach are very important to improve outcome. PMID- 19541694 TI - Pleural electrophysiology variations according to location in pleural cavity. AB - The aim of the study was to compare the electrophysiology profile of sheep pleura originated from different locations of the pleural cavity with the respective profile in humans. Sheep specimens obtained from upper and lower lung lobes, 1st 4th and 8th-12th rib, ventral-dorsal diaphragm and mediastinum were mounted between Ussing chambers. Human visceral tissues were obtained from patients subjected to lobectomy. Trans-mesothelial resistance (R(TM)) was determined as an indicator of the tissue permeability, while amiloride and ouabain were used as inhibitors of cellular transportation via ion transporters. Control values R(TM) were low in lower lobe visceral, caudal costal parietal and diaphragmatic pleura. Amiloride increased R(TM) at all locations except upper visceral and mediastinum. Higher R(TM) increases were found in caudal parietal and dorsal diaphragmatic samples. Ouabain increased R(TM) of lower visceral, caudal parietal and diaphragmatic pleura but not of mediastinal specimens. Observations made in sheep tissue were comparable with human visceral, parietal and mediastinal regions. In conclusion, results suggest heterogeneity of trans-mesothelial permeability among different pleural locations in sheep as was the case for humans. Thoracic surgeons should consider physiology function of each part of pleural cavity before pleural tissue manipulation. Observations made in sheep may be used to understand human physiology. PMID- 19541695 TI - Health systems and increased longevity in people with HIV and AIDS. PMID- 19541696 TI - Risk of pre-eclampsia in first and subsequent pregnancies: prospective cohort study. AB - OBJECTIVE: To investigate whether pre-eclampsia is more common in first pregnancies solely because fewer affected women, who presumably have a higher risk of recurrence, go on to have subsequent pregnancies. DESIGN: Prospective cohort study. SETTING: Swedish Medical Birth Register. PARTICIPANTS: 763 795 primiparous mothers who had their first births in Sweden, 1987-2004. MAIN OUTCOME MEASURES: Pre-eclampsia. RESULTS: The risk of pre-eclampsia was 4.1% in the first pregnancy and 1.7% in later pregnancies overall. However, the risk was 14.7% in the second pregnancy for women who had had pre-eclampsia in their first pregnancy and 31.9% for women who had had pre-eclampsia in the previous two pregnancies. The risk for multiparous women without a history of pre-eclampsia was around 1%. The incidence of pre-eclampsia associated with delivery before 34 weeks' gestation was 0.42% in primiparous women, 0.11% in multiparous women without a history of pre-eclampsia, and 6.8% and 12.5% in women who had had one or two previous pregnancies affected, respectively. The proportion of women who went on to have a further pregnancy was 4-5% lower after having a pregnancy with any pre eclampsia but over 10% lower if pre-eclampsia was associated with very preterm delivery. The estimated risk of pre-eclampsia in parous women did not change with standardisation for pregnancy rates. CONCLUSIONS: Having pre-eclampsia in one pregnancy is a poor predictor of subsequent pregnancy but a strong predictor for recurrence of pre-eclampsia in future gestations. The lower overall risk of pre eclampsia among parous women was not explained by fewer conceptions among women who had had pre-eclampsia in a previous gestation. Early onset pre-eclampsia might be associated with a reduced likelihood of a future pregnancy and with more recurrences than late onset pre-eclampsia when there are further pregnancies. Findings are consistent with the existence of two distinct conditions: a severe recurrent early onset type affected by chronic factors, genetic or environmental, and a milder sporadic form affected by transient factors. PMID- 19541697 TI - Diagnosing and managing vaginismus. PMID- 19541700 TI - Editor's choice: new debate. PMID- 19541701 TI - AIDS epidemic in South Africa may be slowing, survey shows. PMID- 19541702 TI - Specialists recommend counselling women on risks of childbirth after 35. PMID- 19541703 TI - Expert group calls for more funding to tackle malaria. PMID- 19541704 TI - Safety watchdog warns of high dosage errors among children and young people. PMID- 19541705 TI - Is it acceptable for people to take methylphenidate to enhance performance? Yes. PMID- 19541706 TI - Is it acceptable for people to take methylphenidate to enhance performance? No. PMID- 19541707 TI - Effect of withdrawal of co-proxamol on prescribing and deaths from drug poisoning in England and Wales: time series analysis. AB - OBJECTIVE: To assess the effect of the UK Committee on Safety of Medicines' announcement in January 2005 of withdrawal of co-proxamol on analgesic prescribing and poisoning mortality. DESIGN: Interrupted time series analysis for 1998-2007. SETTING: England and Wales. DATA SOURCES: Prescribing data from the prescription statistics department of the Information Centre for Health and Social Care (England) and the Prescribing Services Unit, Health Solutions Wales (Wales). Mortality data from the Office for National Statistics. MAIN OUTCOME MEASURES: Prescriptions. Deaths from drug poisoning (suicides, open verdicts, accidental poisonings) involving single analgesics. RESULTS: A steep reduction in prescribing of co-proxamol occurred in the post-intervention period 2005-7, such that number of prescriptions fell by an average of 859 (95% confidence interval 653 to 1065) thousand per quarter, equating to an overall decrease of about 59%. Prescribing of some other analgesics (co-codamol, paracetamol, co-dydramol, and codeine) increased significantly during this time. These changes were associated with a major reduction in deaths involving co-proxamol compared with the expected number of deaths (an estimated 295 fewer suicides and 349 fewer deaths including accidental poisonings), but no statistical evidence for an increase in deaths involving either other analgesics or other drugs. CONCLUSIONS: Major changes in prescribing after the announcement of the withdrawal of co-proxamol have had a marked beneficial effect on poisoning mortality involving this drug, with little evidence of substitution of suicide method related to increased prescribing of other analgesics. PMID- 19541708 TI - Biomarkers of infection for the differential diagnosis of pleural effusions. AB - We aimed to investigate whether pleural fluid concentrations of biomarkers for bacterial infection, namely triggering receptor expressed on myeloid cells (sTREM 1), procalcitonin (PCT), lipopolysaccharide-binding protein (LBP) and C-reactive protein (CRP), might identify infectious effusions and discriminate between complicated (CPPEs) and uncomplicated parapneumonic effusions (UPPEs). Stored pleural fluid samples from 308 patients with different causes of pleural effusion were used to measure the four biomarkers. Receiver-operating characteristic analysis determined the accuracy of the new tests. Median pleural fluid levels of CRP, sTREM-1 and LBP were significantly higher in CPPE compared with those in other aetiologies. The area under the curve for distinguishing infectious (parapneumonics and tuberculosis) from noninfectious effusions was 0.87 for CRP, 0.86 for sTREM-1, 0.57 for PCT and 0.87 for LBP. Regarding the discrimination of nonpurulent CPPE versus UPPE, a multivariate analysis found that pleural fluid glucose < or =60 mg x dL(-1), LBP > or =17 microg x mL(-1) and CRP > or =80 mg x L(-1) were the best parameters. Individually, none of the new biomarkers achieved better performance characteristics than pH, glucose or lactate dehydrogenase in labelling CPPE. In conclusion, elevated pleural fluid levels of CRP, sTREM and LBP identify patients with infectious effusions, particularly those with CPPE. PCT has no value for the differential diagnosis of pleural effusions. PMID- 19541709 TI - Admission hypoglycaemia is associated with adverse outcome in community-acquired pneumonia. AB - The aim of our study was to investigate if hypoglycaemia correlates with outcome in community-acquired pneumonia (CAP). We performed a prospective, observational study of consecutive patients presenting with a primary diagnosis of CAP in Lothian (UK). Admission plasma glucose was measured and, on this basis, patients were divided into two groups, hypoglycaemic (<4.4 mmol x L(-1) or <79.0 mg x dL( 1)) and nonhypoglycaemic (> or = 4.4 mmol x L(-1) or > or = 79.0 mg x dL(-1)). Outcomes of interest were 30-day mortality, need for mechanical ventilation and inotropic support. Multivariable logistic regression was used to compare these outcomes in hypoglycaemic patients to nonhypoglycaemic patients, adjusting for diabetes mellitus, prior statin use and Pneumonia Severity Index. In total, 1,050 patients were included in the study with 5.4% classified as hypoglycaemic. Increased rates of 30-day mortality (28.1% versus 7.5%, p<0.0001), need for mechanical ventilation (29.8% versus 6.5%, p<0.0001) and need for inotropic support (21.1% versus 4.8%, p<0.0001) were observed in hypoglycaemic patients compared with nonhypoglycaemic patients. On multivariable analysis, hypoglycaemia was independently associated with increased 30-day mortality (OR 2.25, 95% CI 1.1 4.7; p = 0.03), need for mechanical ventilation (OR 3.8, 95% CI 1.9-7.5; p = 0.0002) and need for inotropic support (OR, 2.9, 95% CI 1.4-6.3; p = 0.0006). Admission hypoglycaemia is associated with increased 30-day mortality, need for mechanical ventilation and inotropic support in patients presenting with CAP. PMID- 19541710 TI - Corticosteroid responsiveness and clinical characteristics in childhood difficult asthma. AB - This study describes the clinical characteristics and corticosteroid responsiveness of children with difficult asthma (DA). We hypothesised that complete corticosteroid responsiveness (defined as improved symptoms, normal spirometry, normal exhaled nitric oxide fraction (F(eNO)) and no bronchodilator responsiveness (BDR <12%)) is uncommon in paediatric DA. We report on 102 children, mean+/-sd age 11.6+/-2.8 yrs, with DA in a cross-sectional study. 89 children underwent spirometry, BDR and F(eNO) before and after 2 weeks of systemic corticosteroids (corticosteroid response study). Bronchoscopy was performed after the corticosteroid trial. Of the 102 patients in the cross sectional study, 88 (86%) were atopic, 60 (59%) were male and 52 (51%) had additional or alternative diagnoses. Out of the 81 patients in the corticosteroid response study, nine (11%) were complete responders. Of the 75 patients with symptom data available, 37 (49%) responded symptomatically, which was less likely if there were smokers in the home (OR 0.31, 95% CI 0.02-0.82). Of the 75 patients with available spirometry data, 35 (46%) had normal spirometry, with associations being BAL eosinophilia (OR 5.43, 95% CI 1.13-26.07) and high baseline forced expiratory volume in 1 s (FEV(1)) (OR 1.08, 95% CI 1.02-1.12). Of these 75 patients, BDR data were available in 64, of whom 36 (56%) had <12% BDR. F(eNO) data was available in 70 patients, of whom 53 (75%) had normal F(eNO). Airflow limitation data was available in 75 patients, of whom 17 (26%) had persistent airflow limitation, which was associated with low baseline FEV(1) (OR 0.93, 95% CI 0.90-0.97). Only 11% of DA children exhibited complete corticosteroid responsiveness. The rarity of complete corticosteroid responsiveness suggests alternative therapies are needed for children with DA. PMID- 19541711 TI - Signalling pathways involved in the contractile response to 5-HT in the human pulmonary artery. AB - Serotonin (5-hydroxytryptamine; 5-HT) is a potent pulmonary vasoconstrictor and mitogenic agent whose plasma level is increased in pulmonary hypertensive patients. Thus, we explored the signalling pathways involved in the contractile response to 5-HT in human pulmonary arteries (HPAs). Intact and beta-escin permeabilised rings from HPAs mounted in an organ bath system were used to assess both tension and myofilament Ca(2+)-sensitisation. Microspectrofluorimetry was used for intracellular Ca(2+) recordings in cultured HPA smooth muscle cells. Voltage-operated Ca(2+) channel blockers (nitrendipine and nifedipine) partially reduced the contraction to 5-HT. Thapsigargin or cyclopiazonic acid (CPA), known to deplete sarcoplasmic reticulum Ca(2+) stores, also partially inhibited the contraction, whereas removal of extracellular Ca(2+) under these conditions further inhibited the contraction. Changing from Ca(2+)-free to Ca(2+) containing solution, in the presence of nitrendipine and CPA, a protocol known to stimulate store-operated Ca(2+) channels, induced HPA contractions that were blocked by nickel. Nickel or gadolinium also reduced the contraction to 5-HT. Finally, 5-HT increased intracellular Ca(2+) responses in cultured HPA smooth muscle cells and myofilament Ca(2+)-sensitisation in HPA rings. Collectively, these results indicate that voltage-operated and voltage-independent Ca(2+) channels, as well as Ca(2+) release and myofilament Ca(2+)-sensitisation, participate in 5-HT induced contraction in HPAs. PMID- 19541712 TI - Hyperpolarised 3He MRI versus HRCT in COPD and normal volunteers: PHIL trial. AB - The aim of the present study was to apply hyperpolarised (HP) (3)He magnetic resonance imaging (MRI) to identify patients with chronic obstructive pulmonary disease (COPD) and alpha(1)-antitrypsin deficiency (alpha(1)-ATD) from healthy volunteers and compare HP (3)He MRI findings with high-resolution computed tomography (HRCT) in a multicentre study. Quantitative measurements of HP (3)He MRI (apparent diffusion coefficient (ADC)) and HRCT (mean lung density (MLD)) were correlated with pulmonary function tests. A prospective three centre study enrolled 122 subjects with COPD (either acquired or genetic) and age-matched never-smokers. All diagnostic studies were completed in 94 subjects (52 with COPD; 13 with alpha(1)-ATD; 29 healthy subjects; 63 males; and 31 females; median age 62 yrs). The consensus assessment of radiologists, blinded for other test results, estimated nonventilated lung volume (HP (3)He MRI) and percentage diseased lung (HRCT). Quantitative evaluation of all data for each centre consisted of ADC (HP (3)He MRI) and MLD measurements (HRCT), and correlation with forced expiratory volume in 1 s (FEV(1))/forced vital capacity (FVC) indicating airway obstruction, and the diffusing capacity of the lung for carbon monoxide (D(L,CO)) indicating alveolar destruction. Using lung function tests as a reference, regional analysis of HP (3)He MRI and HRCT correctly categorised normal volunteers in 100% and 97%, COPD in 42% and 69% and alpha(1)-ATD in 69% and 85% of cases, respectively. Direct comparison of HP (3)He MRI and CT revealed 23% of subjects with moderate/severe structural abnormalities had only mild ventilation defects. In comparison with lung function tests, ADC was more effective in separating COPD patients from healthy subjects than MLD (p<0.001 versus 0.038). ADC measurements showed better correlation with D(L,CO) than MLD (r = 0.59 versus 0.29). Hyperpolarised (3)He MRI correctly categorised patients with COPD and normal volunteers. It offers additional functional information, without the use of ionising radiation whereas HRCT gives better morphological information. We showed the feasibility of a multicentre study using different magnetic resonance systems. PMID- 19541713 TI - Does educational level influence lung function decline (Doetinchem Cohort Study)? AB - Low socioeconomic status is associated with reduced lung function in adults. In addition, there are indications that lung function decline with age is accelerated in low socioeconomic groups, but, to date, findings have been inconclusive. In order to investigate the relation between educational level, forced expiratory volume in 1 s (FEV1) and decline in FEV1 over time, linear mixed-effects models were fitted to baseline and 10-yr-follow-up data from the Doetinchem Cohort Study. The study population (26-66 yrs at baseline) consisted of 2,679 males and 3,026 females with an FEV1 measurement in at least one of the three rounds of follow-up and information on relevant covariables. High educational level was used as the reference class. Low educational level was associated with a higher prevalence of smoking and with a lower smoking-adjusted FEV1 at baseline (-148 mL in males and -47 mL in females). In females, low educational level was associated with a faster FEV1 decline (3.4 mL x yr(-1), age and height-adjusted), which was not explained by smoking. In males, no differences in rates of decline between educational levels were observed. FEV1 decline was faster in less-educated females, independent of smoking. In males, FEV1 decline did not differ between educational levels. PMID- 19541714 TI - Control and severity of asthma during pregnancy are associated with asthma incidence in offspring: two-stage case-control study. AB - The extent to which childhood asthma incidence is influenced by asthma control and severity during pregnancy is unknown. We have studied this association during the child's first 10 yrs of life. A two-stage, case-control study, nested in a cohort of 8,226 children of asthmatic mothers, was conducted using three interlinked databases of Quebec, Canada, and mailed questionnaires. A total of 2,681 asthmatic children and 30,318 age-matched controls were selected (< or =20 controls.case(-1); stage 1), and 3,254 selected mothers were mailed questionnaires to obtain additional information (stage 2). Asthma control and severity was defined using validated indexes and childhood asthma incidence based on at least one asthma-related diagnosis and prescription received within 2 yrs. A total of 44 confounders were considered. Compared with children of mild controlled asthmatic mothers, children whose mothers had moderate-to-severe uncontrolled asthma during pregnancy had an increased risk of asthma (adjusted OR 1.27, 95% CI 1.06-1.52). No increased risk was observed for children of mild uncontrolled and moderate-to-severe controlled mothers. Based on one of the largest studies of children of asthmatic mothers, a significant increase in asthma risk was demonstrated among children whose mothers had poor control and increased severity of asthma during pregnancy, indicating that this element should be added to the expanding list of determinants of childhood asthma. As it constitutes a risk factor where pregnant asthmatic females can intervene, it is of great importance for physicians to optimally treat asthmatic females during pregnancy and to encourage females to be adherent to the prescribed asthma medications. PMID- 19541715 TI - Prognostic factors for lung function in systemic sclerosis: prospective study of 105 cases. AB - The aims of the present study were to identify prognostic factors for systemic sclerosis (SSc)-related interstitial lung disease and to clarify the possible causative role of manometric oesophageal involvement. Consecutive SSc patients underwent pulmonary function tests and oesophageal manometry. They were included in the study if pulmonary function tests were repeated >12 months after baseline. The primary end-point was a decrease of >or=10% of the predicted value in forced vital capacity (FVC). The secondary end-points were a decrease of >or=15% pred in lung carbon monoxide diffusing capacity (D(L,CO)) and a decrease of >or=20% pred in FVC. Of the 105 patients (45 diffuse SSc; median disease duration 2.0 yrs), 23 (23%) had a FVC of <80% pred, 60 (59%) had a D(L,CO) of <80% pred and 57 (54%) showed severe oesophageal hypomotility at baseline. Over 72+/-46 months, 29 (28%) patients displayed a decrease of >or=10% pred in FVC, 39 (40%) of 98 patients displayed D(L,CO) decline and 19 (18%) patients displayed a decrease of >or=20% pred in FVC. On multivariate analysis, diffuse SSc was a significant predictor for a decrease of >or=10% pred in FVC (p = 0.01). No other predictor of a decrease in pulmonary function was identified. Only diffuse SSc was predictive of a decrease in pulmonary function in this early-SSc cohort. This does not support preliminary data suggestive of a causative role of oesophageal involvement. PMID- 19541716 TI - Incidence and outcome of weaning from mechanical ventilation according to new categories. AB - Weaning from mechanical ventilation was categorised as simple, difficult or prolonged by an international task force of the American Thoracic Society/European Respiratory Society/European Society of Intensive Care Medicine/Society of Critical Care Medicine/Societe de Reanimation de Langue Francaise in 2007. This new classification has not been tested in clinical practice. The objective of the present study was to determine the incidence and outcome of weaning according to the new categories. We included medical and surgical patients who required mechanical ventilation in a prospective, multicentre, 6-month cohort study. From an initial cohort of 510 patients, 257 intubated patients started weaning. Of these patients, the cumulative incidences of simple, difficult, and prolonged weaning were 152 (59%), 68 (26%) and 37 (14%), respectively. Hospital mortality was increased in patients with prolonged (32%) but not difficult (9%) weaning in comparison with those with simple weaning (13%), overall p = 0.0205. In a multivariate logistic regression model, prolonged but not difficult weaning was associated with an increased risk of death. Ventilator-free days and intensive care unit (ICU)-free days were decreased in both difficult and prolonged weaning. In conclusion, the new weaning category prolonged weaning is associated with increased mortality and morbidity in the ICU. The new category difficult to wean was associated with increased morbidity, but not mortality. PMID- 19541717 TI - A randomised crossover trial of chest physiotherapy in non-cystic fibrosis bronchiectasis. AB - Regular chest physiotherapy is advocated in non-cystic fibrosis bronchiectasis despite little evidence supporting its routine use. This study aimed to establish the efficacy of regular chest physiotherapy in non-cystic fibrosis bronchiectasis compared with no regular chest physiotherapy. 20 patients not practising regular chest physiotherapy were enrolled in a randomised crossover trial of 3 months of twice daily chest physiotherapy using an oscillatory positive expiratory pressure device compared with 3 months of no chest physiotherapy. The primary end-point was the Leicester Cough Questionnaire (LCQ). Additional outcomes included 24-h sputum volume, forced expiratory volume in 1 s (FEV(1)), forced vital capacity (FVC), forced expiratory flow at 25-75% of FVC (FEF(25-75%)), maximum inspiratory pressure (MIP), maximum expiratory pressure (MEP), exercise capacity, sputum microbiology and St George's Respiratory Questionnaire (SGRQ). The treatment effect was estimated using the differences of the pairs of observations from each patient. There was a significant improvement in all domains and total LCQ score with regular chest physiotherapy (median (interquartile range) total score improvement 1.3 (-0.17-3.25) units; p = 0.002). 24-h sputum volume increased significantly with regular chest physiotherapy (2 (0-6) mL; p = 0.02), as did exercise capacity (40 (15-80) m; p = 0.001) and SGRQ total score (7.77 (-0.99 14.5) unit improvement; p = 0.004). No significant differences were seen in sputum bacteriology, FEV(1), FVC, FEF(25-75%), MIP or MEP. Regular chest physiotherapy in non-cystic fibrosis bronchiectasis has small, but significant benefits. PMID- 19541718 TI - Measuring airway dimensions during bronchoscopy using anatomical optical coherence tomography. AB - Airway dimensions are difficult to quantify bronchoscopically because of optical distortion and a limited ability to gauge depth. Anatomical optical coherence tomography (aOCT), a novel imaging technique, may overcome these limitations. This study evaluated the accuracy of aOCT against existing techniques in phantom, excised pig and in vivo human airways. Three comparative studies were performed: 1) micrometer-derived area measurements in 10 plastic tubes were compared with aOCT-derived area; 2) aOCT-derived airway compliance curves from excised pig airways were compared with curves derived using an endoscopic technique; and 3) airway dimensions from the trachea to subsegmental bronchi were measured using aOCT in four anaesthetised patients during bronchoscopy and compared with computed tomography (CT) measurements. Measurements in plastic tubes revealed aOCT to be accurate and reliable. In pig airways, aOCT-derived compliance measurements compared closely with endoscopic data. In human airways, dimensions measured with aOCT and CT correlated closely. Bland-Altman plots showed that aOCT diameter and area measurements were higher than CT measurements by 7.6% and 15.1%, respectively. Airway measurements using aOCT are accurate, reliable and compare favourably with existing imaging techniques. Using aOCT with conventional bronchoscopy allows real-time measurement of airway dimensions and could be useful clinically in settings where knowledge of airway calibre is required. PMID- 19541719 TI - Incidence, characteristics and outcomes of patients with severe community acquired-MRSA pneumonia. AB - We evaluated the published data for the incidence, characteristics and outcomes of patients with community-acquired pneumonia (CAP) due to methicillin-resistant Staphylococcus aureus (MRSA). The estimated incidence of MRSA CAP is 0.51-0.64 cases per 100,000. We identified 74 articles reporting data on 114 patients. Influenza like symptoms was reported in 41% of patients. Pneumonia improved in 59 (54.1%) out of 109 patients; 49 (44.5%) out of 110 patients died. The duration of hospitalisation was 38.1+/-24.9 and 8.3+/-11.7 days, respectively. The duration of intensive care unit (ICU) stay was 18.9+/-13.6 and 6.8+/-9.7 days, respectively. 76 strains carried the Panton-Valentine leukocidin gene. The univariate analysis showed that multi-organ failure (p<0.001), leukopenia (p<0.001), admission to ICU (p<0.001), mechanical ventilation (p<0.001), use of aminoglycosides after culture results (p<0.001), shock (p = 0.001), acute respiratory distress syndrome (p = 0.001), influenza like symptoms (p = 0.008), disseminated intravascular coagulation (p = 0.042) and rash (p = 0.04) were the factors associated with death. PMID- 19541720 TI - Clinical characteristics and prognostic factors of pulmonary MALT lymphoma. AB - Mucosa-associated lymphoid tissue-derived (MALT) lymphoma, a low grade B-cell extranodal lymphoma, is the most frequent subset of primary pulmonary lymphoma. Our objective was to evaluate the initial extent of disease and to analyse the characteristics and long-term outcome of these patients. All chest and pathological departments of teaching hospitals in Paris were contacted in order to identify patients with a histological diagnosis of primary pulmonary lymphoma of the MALT subtype. 63 cases were identified. The median age was 60 yrs. 36% of cases had no symptoms at diagnosis. 46% of patients had at least one extrapulmonary location of lymphoma. The estimated 5- and 10-yr overall survival rates were 90% and 72%, respectively. Only two of the nine observed deaths were related to lymphoma. Age and performance status were the only two adverse prognostic factors for survival. Extrapulmonary location of lymphoma was not a prognostic factor for overall survival or for progression-free survival. Treatment with cyclophosphamide or anthracycline was associated with shorter progression-free survival, when compared with chlorambucil. The survival data confirm the indolent nature of pulmonary MALT lymphoma. Better progression-free survival was observed with chlorambucil when compared with cyclophosphamide or anthracycline. PMID- 19541721 TI - Troponin T predicts in-hospital and 1-year mortality in patients with pulmonary embolism. AB - We aimed to determine the prognostic value of troponin T (TNT) for in-hospital and 1-yr mortality in a large sample of patients with pulmonary embolism (PE). Patients presenting at the emergency department of a tertiary care centre from January 1998 to December 2006 with PE were included. A blood sample was taken at the time of presentation. To determine in-hospital and 1-yr mortality, data from the hospital records and the national death register were used. TNT was determined in 563 out of 737 patients with proven PE. TNT was elevated (>0.03 ng x mL(-1)) in 27%. In-hospital survival was 79% in TNT-positive patients compared with 94% in TNT-negative patients (p<0.001). 1-yr survival was 71% in TNT positive patients compared with 90% in TNT-negative patients (p<0.001). Elevated TNT levels meant a four-times higher risk of in-hospital death and a three-times higher risk of 1-yr mortality, even after adjustment for the other most important risk factors of death in this population. Elevated TNT independently predicts in hospital and 1-yr mortality in patients with acute PE. PMID- 19541722 TI - Multiplex immune serum biomarker profiling in sarcoidosis and systemic sclerosis. AB - Multiplex protein technology has the potential to identify biomarkers for the differentiation, classification and improved understanding of the pathogenesis of interstitial lung disease. The aim of this study was to determine whether a 30 inflammatory biomarker panel could discriminate between healthy controls, sarcoidosis and systemic sclerosis (SSc) patients independently of other clinical indicators. We also evaluated whether a panel of biomarkers could differentiate between the presence or absence of lung fibrosis in SSc patients. We measured 30 circulating biomarkers in 20 SSc patients, 21 sarcoidosis patients and 20 healthy controls using Luminex bead technology and used Fisher's discriminant function analysis to establish the groups of classification mediators. There were significant differences in median concentration measurements between study groups for 20 of the mediators but with considerable range overlap between the groups, limiting group differentiation by single analyte measurements. However, a 17 analyte biomarker model correctly classified 90% of study individuals to their respective group and another 14-biomarker panel correctly identified the presence of lung fibrosis in SSc patients. These findings, if they are corroborated by independent studies in other centres, have potential for clinical application and may generate novel insights into the modulation of immune profiles during disease evolution. PMID- 19541723 TI - Cautious epoprostenol therapy is a safe bridge to lung transplantation in pulmonary veno-occlusive disease. AB - Pulmonary veno-occlusive disease (PVOD) carries a poor prognosis and lung transplantation is the only curative treatment. In PVOD, epoprostenol therapy is controversial, as this condition may be refractory to specific therapy with an increased risk of pulmonary oedema. We retrospectively reviewed clinical, functional and haemodynamic data of 12 patients with PVOD (10 with histological confirmation) treated with continuous intravenous epoprostenol and priority listed for lung transplantation after January 1, 2003. All PVOD patients had severe clinical, functional and haemodynamic impairment at presentation. Epoprostenol was used at low dose ranges with slow dose increases and high dose diuretics. Only one patient developed mild reversible pulmonary oedema. After 3-4 months, improvements were seen in the New York Heart Association functional class (class IV to III in seven patients), cardiac index (1.99+/-0.68 to 2.94+/-0.89 L x min(-1) x m(-2)) and indexed pulmonary vascular resistance (28.4+/-8.4 to 17+/ 5.2 Wood units x m(-2); all p<0.01). A nonsignificant improvement in the 6-min walk distance was also observed (+41 m, p = 0.11). Two patients died, one patient was alive on the transplantation waiting list on December 1, 2008 and nine patients were transplanted. Cautious use of continuous intravenous epoprostenol improved clinical and haemodynamic parameters in PVOD patients at 3-4 months without commonly causing pulmonary oedema, and may be a useful bridge to urgent lung transplantation. PMID- 19541724 TI - Pesticide use and adult-onset asthma among male farmers in the Agricultural Health Study. AB - Although specific pesticides have been associated with wheeze in farmers, little is known about pesticides and asthma. Data from 19,704 male farmers in the Agricultural Health Study were used to evaluate lifetime use of 48 pesticides and prevalent adult-onset asthma, defined as doctor-diagnosed asthma after the age of 20 yrs. Asthma cases were categorised as allergic (n = 127) and nonallergic (n = 314) based on their history of eczema or hay fever. Polytomous logistic regression, controlling for age, state, smoking and body mass, was used to assess pesticide associations. High pesticide exposure events were associated with a doubling of both allergic and nonallergic asthma. For ever-use, 12 individual pesticides were associated with allergic asthma and four with nonallergic asthma. For allergic asthma, coumaphos (OR 2.34; 95% CI 1.49-3.70), heptachlor (OR 2.01; 95% CI 1.30-3.11), parathion (OR 2.05; 95% CI 1.21-3.46), 80/20 mix (carbon tetrachloride/carbon disulfide) (OR 2.15; 95% CI 1.23-3.76) and ethylene dibromide (OR 2.07; 95% CI 1.02-4.20) all showed ORs of >2.0 and significant exposure-response trends. For nonallergic asthma, DDT (dichlorodiphenyltrichloroethane) showed the strongest association (OR 1.41; 95% CI 1.09-1.84), but with little evidence of increasing asthma with increasing use. Current animal handling and farm activities did not confound these results. There was little evidence that allergy alone was driving these associations. In conclusion, pesticides may be an overlooked contributor to asthma risk among farmers. PMID- 19541725 TI - Doing more with less: a conversation with Kerry Weems. AB - The Centers for Medicare and Medicaid Services (CMS) is a resource-starved agency with few friends on Capitol Hill. From the perspective of its last administrator, neither Congress nor the White House, regardless of party, provides CMS with the resources to effectively manage its vast responsibilities-particularly when it comes to fighting fraud, which costs taxpayers billions every year. Although its annual expenditures approach $700 billion, CMS has fewer full-time employees today (about 4,600) than it had a decade ago. With the prospect of health care reform looming, the role that Congress may assign CMS in its weakened state becomes a major policy question. PMID- 19541726 TI - Cytotoxic Activities of Physalis minima L. Chloroform Extract on Human Lung Adenocarcinoma NCI-H23 Cell Lines by Induction of Apoptosis. AB - Physalis minima L. is reputed for having anticancer property. In this study, the chloroform extract of this plant exhibited remarkable cytotoxic activities on NCI H23 (human lung adenocarcinoma) cell line at dose- and time-dependent manners (after 24, 48 and 72 h of incubation). Analysis of cell-death mechanism demonstrated that the extract exerted apoptotic programed cell death in NCI-H23 cells with typical DNA fragmentation, which is a biochemical hallmark of apoptosis. Morphological observation using transmission electron microscope (TEM) also displayed apoptotic characteristics in the treated cells, including clumping and margination of chromatins, followed by convolution of the nuclear and budding of the cells to produce membrane-bound apoptotic bodies. Different stages of apoptotic programed cell death as well as phosphatidylserine externalization were confirmed using annexin V and propidium iodide staining. Furthermore, acute exposure to the extract produced a significant regulation of c-myc, caspase-3 and p53 mRNA expression in this cell line. Due to its apoptotic effect on NCI-H23 cells, it is strongly suggested that the extract could be further developed as an anticancer drug. PMID- 19541727 TI - H1N1 influenza vaccine: Global access for a global problem. PMID- 19541728 TI - Incidence of TNFRSF1A mutations in German children: epidemiological, clinical and genetic characteristics. AB - OBJECTIVE: TNF receptor 1-associated periodic syndrome (TRAPS) is a rare disease belonging to the heterogeneous group of hereditary periodic fever (HPF) syndromes. By their monogenic origins, the HPF syndromes are clearly differentiated from other periodic inflammatory episodes occurring in autoimmune, neoplastic and infectious diseases. We aim to determine the incidence of TRAPS and the spectrum of mutations in the TNFRSF1A gene, and to give a brief survey of clinical signs. METHODS: A prospective surveillance of children with TRAPS was conducted in Germany during a time period of 3 years (2003-06). Monthly inquiries were sent to 370 children's hospitals by the German Pediatric Surveillance Unit (Clinic-ESPED, n1) and to 23 laboratories (Laboratory-ESPED, n2). Inclusion criteria were TNFRSF1A mutation-positive patients < or =16 years of age, more than three self-limiting episodes of fever >38.5 degrees C, and increased inflammation markers. Clinical, epidemiological and genetic data were evaluated via questionnaires. RESULTS: Of the 23 cases included, 19 were identical in 20 clinical and 22 laboratory reports. The incidence of TRAPS in German children was estimated to be approximately 5.6 per 10(7) person-years. In 20 TRAPS patients of the Clinic-ESPED, median age of onset and duration of fever periods were 6 (range 1-16) years and 6.3 (range 2-24) days, respectively. Main symptoms were arthralgia, abdominal pain, lymphadenopathy, headache and skin involvement. The R92Q substitution was found in 19 (83%) of 23 cases. CONCLUSION: The incidence of TRAPS is low and corresponds to 6-10 newly diagnosed patients < or =16 years per year in Germany. PMID- 19541729 TI - Defining disabling foot pain in older adults: further examination of the Manchester Foot Pain and Disability Index. AB - OBJECTIVE: To identify a practical definition of disabling foot pain in older adults for clinical and research use, using the Manchester Foot Pain and Disability Index (FPDI). METHODS: Adults aged > or =50 years registered with three general practices were mailed a two-stage cross-sectional survey. A total of 1342 respondents who reported foot pain in the previous 12 months and completed the FPDI and 58 participants in a test-retest repeatability study were included. RESULTS: Confirmatory factor analysis verified the three-construct FPDI structure (pain intensity, functional limitation and appearance). Internal consistency for the three constructs was good (Cronbach's alpha 0.74, 0.92 and 0.77, respectively). A total of 1320 (98.4%) of those persons with foot pain reported disability (at least one of the 17 FPDI items experienced on at least some days -- Definition A). After restricting this definition to problems experienced on most/every day(s) (Definition B), 996 (74.2%) of those with foot pain reported disability (percentage difference 24.2%; 95% CI 21.9, 26.5%). For each of the three constructs, the prevalence of disability among persons with foot pain was significantly higher under Definition A than under Definition B. Test-retest repeatability for the individual constructs ranged from fair to substantial. Physical function, measured by the SF-36 physical function sub scale, was poorer in those who reported problems within the function construct compared with those with problems in pain and/or appearance constructs only. CONCLUSION: A practical definition of disabling foot pain [at least one of the 10 FPDI function items experienced on most/every day(s)] is proposed, which appears valid, repeatable and suitable for use in older adults. PMID- 19541730 TI - Initial orthostatic hypotension is unrelated to orthostatic tolerance in healthy young subjects. AB - The physiological challenge of standing upright is evidenced by temporary symptoms of light-headedness, dizziness, and nausea. It is not known, however, if initial orthostatic hypotension (IOH) and related symptoms associated with standing are related to the occurrence of syncope. Since IOH reflects immediate and temporary adjustments compared with the sustained adjustments during orthostatic stress, we anticipated that the severity of IOH would be unrelated to syncope. Following a standardized period of supine rest, healthy volunteers [n=46; 25+/-5 yr old (mean+/-SD)] were instructed to stand upright for 3 min, followed by 60 degrees head-up tilt with lower-body negative pressure in 5-min increments of -10 mmHg, until presyncope. Beat-to-beat blood pressure (radial arterial or Finometer), middle cerebral artery blood velocity (MCAv), end-tidal PCO2, and cerebral oxygenation (near-infrared spectroscopy) were recorded continuously. At presyncope, although the reductions in mean arterial pressure, MCAv, and cerebral oxygenation were similar to those during IOH (40+/-11 vs. 43+/ 12%; 36+/-18 vs. 35+/-13%; and 6+/-5 vs. 4+/-2%, respectively), the reduction in end-tidal CO2 was greater (-7+/-6 vs. -4+/-3 mmHg) and was related to the decline in MCAv (R2=0.4; P<0.05). While MCAv pulsatility was elevated with IOH, it was reduced at presyncope (P<0.05). The cardiorespiratory and cerebrovascular changes during IOH were unrelated to those at presyncope, and interestingly, there was no relationship between the hemodynamic changes and the incidence of subjective symptoms in either scenario. During IOH, the transient nature of physiological changes can be well tolerated; however, potentially mediated by a reduced MCAv pulsatility and greater degree of hypocapnic-induced cerebral vasoconstriction, when comparable changes are sustained, the development of syncope is imminent. PMID- 19541731 TI - Rat Achilles tendon healing: mechanical loading and gene expression. AB - Injured tendons require mechanical tension for optimal healing, but it is unclear which genes are upregulated and responsible for this effect. We unloaded one Achilles tendon in rats by Botox injections in the calf muscles. The tendon was then transected and left to heal. We studied mechanical properties of the tendon calluses, as well as mRNA expression, and compared them with loaded controls. Tendon calluses were studied 3, 8, 14, and 21 days after transection. Intact tendons were studied similarly for comparison. Altogether 110 rats were used. The genes were chosen for proteins marking inflammation, growth, extracellular matrix, and tendon specificity. In intact tendons, procollagen III and tenascin-C were more expressed in loaded than unloaded tendons, but none of the other genes was affected. In healing tendons, loading status had small effects on the selected genes. However, TNF-alpha, transforming growth factor-beta1, and procollagens I and III were less expressed in loaded callus tissue at day 3. At day 8 procollagens I and III, lysyl oxidase, and scleraxis had a lower expression in loaded calluses. However, by days 14 and 21, procollagen I, cartilage oligomeric matrix protein, tenascin-C, tenomodulin, and scleraxis were all more expressed in loaded calluses. In healing tendons, the transverse area was larger in loaded samples, but material properties were unaffected, or even impaired. Thus mechanical loading is important for growth of the callus but not its mechanical quality. The main effect of loading during healing might thereby be sought among growth stimulators. In the late phase of healing, tendon-specific genes (scleraxis and tenomodulin) were upregulated with loading, and the healing tissue might to some extent represent a regenerate rather than a scar. PMID- 19541732 TI - Three-dimensional measurement of alveolar airspace volumes in normal and emphysematous lungs using micro-CT. AB - In pulmonary emphysema, the alveolar structure progressively breaks down via a three-dimensional (3D) process that leads to airspace enlargement. The characterization of such structural changes has, however, been based on measurements from two-dimensional (2D) tissue sections or estimates of 3D structure from 2D measurements. In this study, we developed a novel silver staining method for visualizing tissue structure in 3D using micro-computed tomographic (CT) imaging, which showed that at 30 cmH20 fixing pressure, the mean alveolar airspace volume increased from 0.12 nl in normal mice to 0.44 nl and 2.14 nl in emphysematous mice, respectively, at 7 and 14 days following elastase induced injury. We also assessed tissue structure in 2D using laser scanning confocal microscopy. The mean of the equivalent diameters of the alveolar airspaces was lower in 2D compared with 3D, while its variance was higher in 2D than in 3D in all groups. However, statistical comparisons of alveolar airspace size from normal and emphysematous mice yielded similar results in 2D and 3D: compared with control, both the mean and variance of the equivalent diameters increased by 7 days after treatment. These indexes further increased from day 7 to day 14 following treatment. During the first 7 days following treatment, the relative change in SD increased at a much faster rate compared with the relative change in mean equivalent diameter. We conclude that quantifying heterogeneity in structure can provide new insight into the pathogenesis or progression of emphysema that is enhanced by improved sensitivity using 3D measurements. PMID- 19541733 TI - Effect of compliance and hematocrit on wall shear stress in a model of the entire coronary arterial tree. AB - A hemodynamic analysis is implemented in the entire coronary arterial tree of diastolically arrested, vasodilated pig heart that takes into account vessel compliance and blood viscosity in each vessel of a large-scale simulation involving millions of vessels. The feed hematocrit (Hct) is varied at the inlet of the coronary arterial tree, and the Fahraeus-Lindqvist effect and phase separation are considered throughout the vasculature. The major findings are as follows: 1) vessel compliance is the major determinant of nonlinearity of the pressure-flow relation, and 2) changes in Hct influence wall shear stress (WSS) in epicardial coronary arteries more significantly than in transmural and perfusion arterioles because of the Fahraeus-Lindqvist effect. The present study predicts the flow rate as a second-order polynomial function of inlet pressure due to vessel compliance. WSS in epicardial coronary arteries increases >15% with an increase of feed Hct from 45% to 60% and decreases >15% with a decrease of feed Hct from 45% to 30%, whereas WSS in small arterioles is not affected as feed Hct changes in this range. These findings have important implications for acute Hct changes under vasodilated conditions. PMID- 19541734 TI - Evaluation of individual skeletal muscle activity by glucose uptake during pedaling exercise at different workloads using positron emission tomography. AB - Skeletal muscle glucose uptake closely reflects muscle activity at exercise intensity levels <55% of maximal oxygen consumption (VO2max). Our purpose was to evaluate individual skeletal muscle activity from glucose uptake in humans during pedaling exercise at different workloads by using [18F]fluorodeoxyglucose (FDG) and positron emission tomography (PET). Twenty healthy male subjects were divided into two groups (7 exercise subjects and 13 control subjects). Exercise subjects were studied during 35 min of pedaling exercise at 40 and 55% VO2max exercise intensities. FDG was injected 10 min after the start of exercise or after 20 min of rest. PET scanning of the whole body was conducted after completion of the exercise or rest period. In exercise subjects, mean FDG uptake [standardized uptake ratio (SUR)] of the iliacus muscle and muscles of the anterior part of the thigh was significantly greater than uptake in muscles of control subjects. At 55% VO2max exercise, SURs of the iliacus muscle and thigh muscles, except for the rectus femoris, increased significantly compared with SURs at 40% VO2max exercise. Our results are the first to clarify that the iliacus muscle, as well as the muscles of the anterior thigh, is the prime muscle used during pedaling exercise. In addition, the iliacus muscle and all muscles in the thigh, except for the rectus femoris, contribute when the workload of the pedaling exercise increases from 40 to 55% VO2max. PMID- 19541735 TI - Tissue memory in healing tendons: short loading episodes stimulate healing. AB - Intact tendons adapt slowly to changes in mechanical loading, whereas in healing tendons the effect of mechanical loading or its absence is dramatic. The longevity of the response to a single loading episode is, however, unknown. We hypothesized that the tissue has a "memory" of loading episodes and that therefore short loadings are sufficient to elicit improved healing. The Achilles tendon of 70 female rats was transected and unloaded by tail suspension for 12 days (suspension started on day 2 after surgery). Each day, the rats were let down from suspension for short daily training episodes according to different regimes: 15 min of cage activity or treadmill running for 15, 30, 60, or 2x15 min. Rats with transected Achilles tendons and full-time cage activity served as controls. The results demonstrated that full-time cage activity increased the peak force over three times compared with unloading. Short daily loading episodes (treadmill running) increased the peak force about half as much as full-time activity. Prolongation of treadmill running above 15 min or dividing the daily training in two separate episodes had minimal further effect. This mechanical stimulation increased the cross-sectional area but had no effect on the mechanical properties of the repair tissue. The findings indicate that once the tissue had received information from a certain loading type and level, this is "memorized" and leads to a response lasting many hours. This suggests that patients might be allowed early short loading episodes following, e.g., an Achilles tendon rupture for a better outcome. PMID- 19541736 TI - Bronchodilator effects of exercise hyperpnea and albuterol in mild-to-moderate asthma. AB - In asthmatic patients, either bronchodilatation or bronchoconstriction may develop during exercise. In 18 patients with mild-to-moderate asthma, we conducted two studies with the aims to 1) quantify the bronchodilator effect of hyperpnea induced by incremental-load maximum exercise compared with effects of inhaled albuterol (study 1, n=10) and 2) determine the time course of changes in airway caliber during prolonged constant-load exercise (study 2, n=8). In both studies, it was also investigated whether the bronchodilator effects of exercise hyperpnea and albuterol are additive. Changes in airway caliber were measured by changes in partial forced expiratory flow. In study 1, incremental-load exercise was associated with a bronchodilatation that was approximately 60% of the maximal bronchodilatation obtainable with 1,500 microg of albuterol. In study 2, constant load exercise was associated with an initial moderate bronchodilatation and a late airway renarrowing. In both studies, premedication with inhaled albuterol (400 microg) promoted sustained bronchodilatation during exercise, which was additive to that caused by exercise hyperpnea. In conclusion, in mild-to-moderate asthmatic individuals, hyperpnea at peak exercise was associated with a potent yet not complete bronchodilatation. During constant-load exercise, a transient bronchodilatation was followed by airway renarrowing, suggesting prevalence of constrictor over dilator effects of hyperpnea. Finally, the bronchodilator effect of hyperpnea was additive to that of albuterol. PMID- 19541738 TI - Comparison of regional patch collection vs. whole body washdown for measuring sweat sodium and potassium loss during exercise. AB - This study compared simultaneous whole body washdown (WBW) and regional skin surface (REG) sweat collections to generate regression equations to predict WBW sweat Na(+) concentration ([Na(+)]) and K(+) concentration ([K(+)]) from single- and five-site REG sweat patch collections. Athletes (10 men, 10 women) cycled in a plastic chamber for 90 min in the heat. Before exercise, the subject and bike were washed with deionized water. After the onset of sweating, sterile patches were attached to the forearm, back, chest, forehead, and thigh and removed on saturation. After exercise, the subject and bike were washed with ammonium sulfate solution to collect all sweat electrolyte loss and determine the volume of unevaporated sweat. All individual patch sites and five-site REG (weighted for local sweat rate and body surface area) were significantly (P = 0.000) correlated with WBW sweat [Na(+)]. The equation for predicting WBW sweat [Na(+)] from five site REG was y = 0.68x + 0.44 [r = 0.97, intraclass correlation coefficient (ICC) = 0.70] and did not differ between sexes. There were sex differences in the regression results between five-site REG and WBW sweat [K(+)] (men: y = 0.74x + 0.30, r = 0.89, ICC = 0.73; women: y = 0.04x + 3.18, r = 0.03, ICC = 0.00). Five site REG sweat [Na(+)] and [K(+)] significantly overestimated that of WBW sweat (59 +/- 27 vs. 41 +/- 19 meq/l, P = 0.000 and 4.4 +/- 0.7 vs. 3.6 +/- 0.7 meq/l, P = 0.000, respectively). For both sexes, the best sites for predicting WBW sweat [Na(+)] and [K(+)] were the thigh (1 +/- 8 meq/l < WBW, P = 1.000, y = 0.75x + 11.37, r = 0.96, ICC = 0.93) and chest (0.2 +/- 0.3 meq/l > WBW, P = 1.000, y = 0.76x + 0.55, r = 0.89, ICC = 0.87), respectively. In conclusion, regression equations can be used to accurately and reliably predict WBW sweat [Na(+)] and [K(+)] from REG sweat collections when study conditions and techniques are similar to that of the present protocol. PMID- 19541737 TI - Sympathetic neural responses to mental stress during acute simulated microgravity. AB - Neural and cardiovascular responses to mental stress and acute 6 degrees head down tilt (HDT) were examined separately and combined. We hypothesized sympathoexcitation during mental stress, sympathoinhibition during HDT, and an additive neural interaction during combined mental stress and HDT. Muscle sympathetic nerve activity (MSNA), mean arterial pressure (MAP), and heart rate (HR) were recorded in 16 healthy subjects (8 men, 8 women) in the supine position during three randomized trials: 1) mental stress (via mental arithmetic), 2) HDT, and 3) combined mental stress and HDT. Mental stress significantly increased MSNA (7+/-1 to 12+/-2 bursts/min; P<0.01), MAP (91+/-2 to 103+/-2 mmHg; P<0.01), and HR (70+/-3 to 82+/-3 beats/min; P<0.01). HDT did not change MSNA or HR, but MAP was reduced (91+/-2 to 89+/-3 mmHg; P<0.05). Combined mental stress and HDT significantly increased MSNA (7+/-1 to 10+/-1 bursts/min; P<0.01), MAP (88+/-3 to 99+/-3 mmHg; P<0.01), and HR (70+/-3 to 82+/-3 beats/min; P<0.01). Increases in MSNA and HR during the combination trial were not different from the sum of the individual trials. However, the increase in MAP during the combination trial was significantly greater than the sum of the individual trials (change of 11+/-1 vs. 9+/-1 mmHg; P<0.05). We conclude that the interaction for MSNA and HR are additive during combined mental stress and HDT but that MAP responses are slightly augmented during the combined trial. These findings demonstrate that sympathetic neural responses to mental stress are unaltered by simulated microgravity. PMID- 19541739 TI - The fastest runner on artificial legs: different limbs, similar function? AB - The recent competitive successes of a bilateral, transtibial amputee sprint runner who races with modern running prostheses has triggered an international controversy regarding the relative function provided by his artificial limbs. Here, we conducted three tests of functional similarity between this amputee sprinter and competitive male runners with intact limbs: the metabolic cost of running, sprinting endurance, and running mechanics. Metabolic and mechanical data, respectively, were acquired via indirect calorimetry and ground reaction force measurements during constant-speed, level treadmill running. First, we found that the mean gross metabolic cost of transport of our amputee sprint subject (174.9 ml O(2)*kg(-1)*km(-1); speeds: 2.5-4.1 m/s) was only 3.8% lower than mean values for intact-limb elite distance runners and 6.7% lower than for subelite distance runners but 17% lower than for intact-limb 400-m specialists [210.6 (SD 13.2) ml O(2)*kg(-1)*km(-1)]. Second, the speeds that our amputee sprinter maintained for six all-out, constant-speed trials to failure (speeds: 6.6-10.8 m/s; durations: 2-90 s) were within 2.2 (SD 0.6)% of those predicted for intact-limb sprinters. Third, at sprinting speeds of 8.0, 9.0, and 10.0 m/s, our amputee subject had longer foot-ground contact times [+14.7 (SD 4.2)%], shorter aerial [-26.4 (SD 9.9)%] and swing times [-15.2 (SD 6.9)%], and lower stance averaged vertical forces [-19.3 (SD 3.1)%] than intact-limb sprinters [top speeds = 10.8 vs. 10.8 (SD 0.6) m/s]. We conclude that running on modern, lower-limb sprinting prostheses appears to be physiologically similar but mechanically different from running with intact limbs. PMID- 19541740 TI - Relationship between body mass index and adiposity in prepubertal children: ethnic and geographic comparisons between New York City and Jinan City (China). AB - Body mass index (BMI) is often used as a surrogate estimate of percent body fat in epidemiological studies. This study tested the hypothesis that BMI is representative of body fatness independent of age, sex, ethnicity, and geographic location in prepubertal children. The study sample included a total of 605 prepubertal children (275 girls and 330 boys) of which 247 were Chinese from Jinan, Shandong, Mainland China, and 358 children were from various ethnic backgrounds in New York City (NYC): 121 Caucasians, 94 African Americans, and 143 Asians (Chinese and Korean). In this cross-sectional study, dual energy X-ray absorptiometry was used to quantify total body fat (TBF) and percent body fat (PBF). Prepubertal status was assessed by the criteria of Tanner. Multiple regression models were developed with TBF and PBF as the dependent variables and BMI, age, sex, and ethnicity as independent variables. Multiple regression analysis showed that BMI alone explained 85% and 69% of between-subject variance for TBF and PBF, respectively. Sex was a significant contributor to the models (P<0.001) with girls having higher TBF and PBF than boys. Ethnicity and geographic location were significant contributors to the model (P<0.0001) with Asians (Jinan and NYC Asians) having higher PBF than all non-Asian groups (P<0.0001), and Jinan Asians having higher TBF and PBF than NYC-Asians. Among prepubertal children, for the same BMI, Asians have significantly higher PBF compared with African Americans and Caucasians. Caution is warranted when applying BMI across sex and ethnic prepubertal groups. PMID- 19541741 TI - Mechanical effects of obesity on airway responsiveness in otherwise healthy humans. AB - We investigated whether obesity is associated with airway hyperresponsiveness in otherwise healthy humans and, if so, whether this correlates with a restrictive lung function pattern or a decreased number of sighs at rest and/or during walking. Lung function was studied before and after inhaling methacholine (MCh) in 41 healthy subjects with body mass index ranging from 20 to 56. Breathing pattern was assessed during a 60-min rest period and a 30-min walk. The dose of MCh that produced a 50% decrease in the maximum expiratory flow measured in a body plethysmograph (PD50MCh) was inversely correlated with body mass index (r2=0.32, P<0.001) and waist circumference (r2=0.25, P<0.001). Significant correlations with body mass index were also found with the maximum changes in respiratory resistance (r2=0.19, P<0.001) and reactance (r2=0.40, P<0.001) measured at 5 Hz. PD50MCh was also positively correlated with functional residual capacity (r2=0.56, P<0.001) and total lung capacity (r2=0.59, P<0.001) in men, but not in women. Neither PD50MCh nor body mass index correlated with number of sighs, average tidal volume, ventilation, or breathing frequency. In this study, airway hyperresponsiveness was significantly associated with obesity in otherwise healthy subjects. In obese men, but not in women, airway hyperresponsiveness was associated with the decreases in lung volumes. PMID- 19541742 TI - In vivo strain of the medial vs. lateral quadriceps tendon in patellofemoral pain syndrome. AB - Patellofemoral pain (PFP) is thought to be related to patellar maltracking due to imbalances in the knee extensor. However, no study has evaluated the in vivo biomechanical properties of the quadriceps tendon in PFP syndrome. Our purpose was to compare the biomechanical properties of the quadriceps tendons in vivo and noninvasively in patients with PFP syndrome to those of control subjects. The null hypothesis was that the quadriceps tendons of PFP subjects would have significantly decreased strain compared with control subjects. Fourteen subjects (7 control, 7 PFP) performed voluntary ramp isometric contractions to a range of torque levels, while quadriceps tendon elongation was measured using ultrasonography. Tendon strain was calculated for the vastus medialis obliquus (VMO) and vastus lateralis (VL) portion of the quadriceps tendon and compared between subjects (control vs. PFP) and within subjects (VMO vs. VL). PFP subjects showed significantly less VMO tendon strain than control subjects (P<0.001), but there was no difference in VL tendon strain between PFP and control subjects (P=0.100). Relative weakness of the VMO is the most likely cause of the decreased tendon strain seen in subjects with PFP. VMO weakness not only explains the decreased medial tendon strain but also explains the presence of increased lateral patellar translation and lateral patellar spin (distal pole rotates laterally) reported in the literature in this population. This technique can potentially be used in a clinical setting to evaluate quadriceps tendon properties and infer the presence of muscle weakness in PFP. PMID- 19541743 TI - Minireview: Pref-1: role in adipogenesis and mesenchymal cell fate. AB - Preadipocyte factor-1 [Pref-1; also called Dlk1 (Delta-like protein 1)] is made as an epidermal growth factor-repeat containing transmembrane protein that produces a biologically active soluble form by TNF-alpha-converting enzyme (TACE) mediated cleavage. Soluble Pref-1 activates the MAPK kinase/ERK pathway. In adipose tissue, Pref-1 is specifically expressed in preadipocytes but not in adipocytes and thus is used as a preadipocyte marker. Inhibition of adipogenesis by Pref-1 has been well established in vitro as well as in vivo by ablation and overexpression of Pref-1. SRY (sex determining region Y)-box 9 (Sox9), a transcription factor expressed in preadipocytes to suppress CCAAT enhancer binding protein beta and (C/EBP) delta expression, is required to be down regulated before adipocyte differentiation can proceed. By activating MAPK kinase/ERK, Pref-1 prevents down-regulation of Sox9, resulting in inhibition of adipogenesis. Furthermore, by inducing Sox9, Pref-1 promotes chondrogenic induction of mesenchymal cells but prevents chondrocyte maturation as well as osteoblast differentiation. Thus, Pref-1 directs multipotent mesenchymal cells toward the chondrogenic lineage but inhibits differentiation into adipocytes as well as osteoblasts and chondrocytes. Pref-1, encoded by an imprinted gene, has also been detected in progenitor cells in various tissues during regeneration and therefore may have a more general role in maintaining cells in an undifferentiated state. PMID- 19541744 TI - Structural and functional characterization of the interdomain interaction in the mineralocorticoid receptor. AB - The mineralocorticoid receptor (MR) plays a central role in electrolyte homeostasis and in cardiovascular disease. We have previously reported a ligand dependent N/C-interaction in the MR. In the present study we sought to fully characterize the MR N/C-interaction. By using a range of natural and synthetic MR ligands in a mammalian two-hybrid assay we demonstrate that in contrast to aldosterone, which strongly induces the interaction, the physiological ligands deoxycorticosterone and cortisol weakly promote the interaction but predominantly inhibit the aldosterone-mediated N/C-interaction. Similarly, progesterone and dexamethasone antagonize the interaction. In contrast, the synthetic agonist 9alpha-fludrocortisol robustly induces the interaction. The ability of the N/C interaction to discriminate between MR agonists suggests a subtle conformational difference in the ligand-binding domain induced by these agonists. We also demonstrate that the N/C interaction is not cell specific, consistent with the evidence from a glutathione-S-transferase pull-down assay, of a direct protein protein interaction between the N- and C-terminal domains of the MR. Examination of a panel of deletions in the N terminus suggests that several regions may be critical to the N/C-interaction. These studies have identified functional differences between physiological MR ligands, which suggest that the ligand specific dependence of the N/C-interaction may contribute to the differential activation of the MR that has been reported in vivo. PMID- 19541745 TI - Subcellular trafficking of the TRH receptor: effect of phosphorylation. AB - Activation of the G protein-coupled TRH receptor leads to its phosphorylation and internalization. These studies addressed the fundamental question of whether phosphorylation regulates receptor trafficking or endosomal localization regulates the phosphorylation state of the receptor. Trafficking of phosphorylated and dephosphorylated TRH receptors was characterized using phosphosite-specific antibody after labeling surface receptors with antibody to an extracellular epitope tag. Rab5 and phosphoreceptor did not colocalize at the plasma membrane immediately after TRH addition but overlapped extensively by 15 min. Dominant-negative Rab5-S34N inhibited receptor internalization. Later, phosphoreceptor was in endosomes containing Rab5 and Rab4. Dephosphorylated receptor colocalized with Rab4 but not with Rab5. Dominant-negative Rab4, -5, or 11 did not affect receptor phosphorylation or dephosphorylation, showing that phosphorylation determines localization in Rab4(+)/Rab5(-) vesicles and not vice versa. No receptor colocalized with Rab7; a small amount of phosphoreceptor colocalized with Rab11. To characterize recycling, surface receptors were tagged with antibody, or surface receptors containing an N-terminal biotin ligase acceptor sequence were labeled with biotin. Most recycling receptors did not return to the plasma membrane for more than 2 h after TRH was removed, whereas the total cell surface receptor density was largely restored in less than 1 h, indicating that recruited receptors contribute heavily to early repopulation of the plasma membrane. PMID- 19541746 TI - Dual ablation of Grb10 and Grb14 in mice reveals their combined role in regulation of insulin signaling and glucose homeostasis. AB - Growth factor receptor bound (Grb)10 and Grb14 are closely related adaptor proteins that bind directly to the insulin receptor (IR) and regulate insulin induced IR tyrosine phosphorylation and signaling to IRS-1 and Akt. Grb10- and Grb14-deficient mice both exhibit improved whole-body glucose homeostasis as a consequence of enhanced insulin signaling and, in the case of the former, altered body composition. However, the combined physiological role of these adaptors has remained undefined. In this study we utilize compound gene knockout mice to demonstrate that although deficiency in one adaptor can enhance insulin-induced IRS-1 phosphorylation and Akt activation, insulin signaling is not increased further upon dual ablation of Grb10 and Grb14. Context-dependent limiting mechanisms appear to include IR hypophosphorylation and decreased IRS-1 expression. In addition, the compound knockouts exhibit an increase in lean mass comparable to Grb10-deficient mice, indicating that this reflects a regulatory function specific to Grb10. However, despite the absence of additive effects on insulin signaling and body composition, the double-knockout mice are protected from the impaired glucose tolerance that results from high-fat feeding, whereas protection is not observed with animals deficient for individual adaptors. These results indicate that, in addition to their described effects on IRS-1/Akt, Grb10 and Grb14 may regulate whole-body glucose homeostasis by additional mechanisms and highlight these adaptors as potential therapeutic targets for amelioration of the insulin resistance associated with type 2 diabetes. PMID- 19541747 TI - Functional characterization of a promoter polymorphism in APE1/Ref-1 that contributes to reduced lung cancer susceptibility. AB - Apurinic/apyrimidinic endonuclease 1/redox effector factor-1 (APE1/Ref-1) is a ubiquitous multifunctional protein that possesses both DNA-repair and redox regulatory activities. Although it was originally identified as a DNA-repair enzyme, accumulating evidence supports a role of APE1/Ref-1 in tumor development. To investigate association between APE1/Ref-1 polymorphisms and lung cancer risk in Chinese populations, we first genotyped three variants of APE1/Ref-1 and found a -141 T-to-G variant (rs1760944) in the promoter associated with decreased risk of lung cancer [odds ratio (OR) = 0.62 for GG; P=0.043]. Similar results were obtained in a follow-up replication study. Combined data from the two studies comprising a total of 1072 lung cancer patients and 1064 cancer-free control participants generated a more significant association (P=0.002). We observed lower APE1/Ref-1 mRNA levels in the presence of the protective G allele in human peripheral blood mononuclear cells and normal lung tissues. The -141G-allele promoter construct exhibited decreased luciferase reporter gene expression. Electrophoretic mobility shift assays and surface plasmon resonance analysis showed that the -141G allele impaired the binding affinity of some transcription factor, accounting for lower APE1/Ref-1-promoter activity. Supershift assays further revealed that the protein of interest was octamer-binding transcription factor-1 (Oct-1). Chromatin immunoprecipitation reconfirmed binding of Oct-1 to the APE1/Ref-1 -141-promoter region. We also found that Oct-1 conferred attenuated transactivation capacity toward the -141G variant by exogenously introducing Oct-1. These data indicate that genetic variations in APE1/Ref-1 may modify susceptibility to lung cancer and provide new insights into an unexpected effect of APE1/Ref-1 on lung carcinogenesis. PMID- 19541748 TI - The validity and clinical utility of post-traumatic stress disorder in Afghanistan. AB - This study examined the validity and utility of PTSD among 320 adults in Afghanistan. Findings support the validity of PTSD in this cultural context: PTSD symptoms were highly prevalent, shared common variance, and correlated as expected with exposure to traumatic stress. However, only limited support was found for the clinical utility of PTSD. Other types of psychiatric symptomatology, including depression and a culturally specific measure of general distress, correlated more highly with traumatic stress than did PTSD; and PTSD accounted for limited variance in functioning beyond that explained by depression and general distress. Implications for research and intervention are considered. PMID- 19541749 TI - A qualitative study of mental health problems among children displaced by war in northern Uganda. AB - While multiple studies have found that children affected by war are at increased risk for a range of mental health problems, little research has investigated how mental health problems are perceived locally. In this study we used a previously developed rapid ethnographic assessment method to explore local perceptions of mental health problems among children and adults from the Acholi ethnic group displaced by the war in northern Uganda. We conducted 45 free list interviews and 57 key informant interviews. The rapid assessment approach appears to have worked well for interviewing caretakers and children aged 10-17 years. We describe several locally defined syndromes: two tam/par/kumu (depression and dysthymia like syndromes), ma lwor (a mixed anxiety and depression-like syndrome), and a category of conduct problems referred to as kwo maraco/gin lugero. The descriptions of these local syndromes were similar to western mood, anxiety and conduct disorders, but included culture-specific elements. PMID- 19541750 TI - Immigrants' continuing bonds with their native culture: assimilation analysis of three interviews. AB - Three case studies of immigrants to the US from China, Iraq, and Mexico were used to build a theory of acculturation in immigrants by integrating the continuing bonds model, which describes mourning in bereavement with the assimilation model, which describes psychological change in psychotherapy. Participants were interviewed about the loss of their native culture and their life in the US. One participant had not fully assimilated the loss of her native culture, but used her continuing bonds with her culture as a source of solace. Another participant used his continuing bonds with his culture as a source of solace, but these bonds had become a source of conflict with the host culture. The third participant had largely assimilated the loss of his native culture such that the voices of this culture were linked via meaning bridges with the voices of the host culture, and the continuing bonds were resources that helped him in his land of immigration. PMID- 19541751 TI - Problemas de nervos: a multivocal symbol of distress for Portuguese immigrants. AB - This article outlines research on a previous unstudied form of suffering specific to the Portugese immigrant community: problemas de nervos. Thirty-two Portuguese immigrant women (in Waterloo, ON and Boston, MA) were interviewed and each completed a questionnaire. Cluster analysis demonstrated that problemas de nervos has many meanings. The study profiled symptoms, causes and therapies associated with four variations of this culture-specific form of distress: "mal da cabeca" meaning problems with/in the head (e.g., lack of control, visions); " aflicao" meaning affliction (e.g., nervous attacks, heart problems); immigration stress (causing sleep disturbances); and, conflicts with others (resulting in pressure within the body). None of the symptom clusters reported matched criteria for a DSM-IV-TR diagnosis, suggesting that problemas de nervos represents an idiomatic rather than universal expression of distress. PMID- 19541752 TI - What research tells us about the mental health and psychosocial wellbeing of Sudanese refugees: a literature review. AB - Sudan has long been one of the world's chief refugee-producing nations. Many researchers and practitioners have developed considerable interest in culturally specific information on the mental health and wellbeing of Sudanese refugees. In this selective review of studies with Sudanese refugees, on mental health and psychosocial wellbeing, coping strategies and interventions, most quantitative studies found high rates of psychopathology, particularly PTSD and depression. However, some studies using mixed methods cautioned that while many Sudanese refugees have symptoms of traumatic stress, their functioning was not necessarily reduced, and they themselves often reported more concern with current stressors such as family problems than with past trauma. Some qualitative studies suggest that many Sudanese refugees use coping strategies such as silence, stoicism, and suppression. Few studies were available regarding appropriate interventions for Sudanese refugees and it remains unclear which aspects of standard treatments used by western-trained mental health practitioners may be beneficial for members of this population. PMID- 19541753 TI - Working together for public health. AB - Italy's recent economic growth and strategic position in the Mediterranean Sea have made it a prime destination for immigrants and asylum seekers in Europe. Despite its well-developed health care system, statistics on foreign citizens' health are worrisome. In 1998 public health services were extended to illegal immigrants, giving them the right to necessary urgent and non-urgent medical assistance, even for a prolonged period. This paper examines a two-year joint intervention project between Centre for the Study and Research of Public Health (Mental Health), Local Health Agency ROMA E (LHA RME) and the non-governmental organization Medecins Sans Frontieres (MSF) in Rome. PMID- 19541754 TI - The role of leisure in the lives of psychotic patients: a qualitative study. AB - Quality of life is defined by indicators that measure the life patterns of a person or community in relation to an ideal model. Leisure is an important component of quality of life and a tool for rehabilitation for patients with chronic psychosis. This qualitative study evaluated the role of leisure in the life of patients living with psychosis. Interviews with patients and relatives indicate that sociability is an important value for both groups. Deeper analysis, however, indicates that sociability is a culturally learned concept for patients and does not relate to their real needs. Improvement of symptoms and respect of individuality are more central to their well-being. PMID- 19541755 TI - Autism in Saudi Arabia: presentation, clinical correlates and comorbidity. AB - Although autism is a global disorder, relatively little is known about its presentation and occurrence in many developing countries, such as Saudi Arabia. This article describes the reasons for referral and clinical characteristics of a sample of 49 children (37 males and 12 females) diagnosed with an autistic spectrum disorder at a tertiary referral center in Saudi Arabia. The diagnosis of autism was based on DSM-IV criteria supplemented by information obtained from parent and child interviews, rating scales, and examination of school and hospital records. Females were older than males at the time of referral. Eleven patients had a history of seizure disorder and one patient had a chromosome abnormality. Twenty-five patients were taking psychotropic medications and 14 patients were the product of consanguineous marriages. Clinical and research implications of these findings are discussed. PMID- 19541756 TI - Strange, incredible and impossible things: the early anthropology of Reginald Scot. AB - Reginald Scot has been acclaimed as an early rationalist for his critical consideration of witchcraft in 1584. At the same time, the Discoverie of Witchcraft appears organized much as later classic anthropological monographs. This article considers whether his methods and writing might indeed correspond to what we recognise as the procedures of medical or psychiatric anthropology. PMID- 19541757 TI - Mass possession state in a family setting. PMID- 19541758 TI - The cultural myth of Koro conceptualization: time for a rethink. PMID- 19541759 TI - The prevalence of problematic gambling behaviour: a Scandinavian comparison. AB - AIMS: On the basis of a new large-scale screening for the prevalence of problematic gambling behaviour in Denmark, the aim of the study was: (a) to evaluate the effect of applying different screening tools; and (b) to compare gambling behaviour in Denmark with the prevalence of problematic gambling behaviour in other Scandinavian countries. METHODS: The screening tools applied were the internationally validated SOGS-R and NODS. These were used within the same survey, thus allowing for comparison with surveys conducted in other Scandinavian countries where either the one or the other tool has been utilized. RESULTS: The prevalence of at-risk gamblers, problematic gamblers and pathological gamblers was significantly lower in Denmark than in the other Scandinavian countries. This held true for both genders as well as for different age groups in comparisons of Denmark and Norway. CONCLUSIONS: There is a considerable variation in the prevalence of problematic gambling between the Scandinavian countries, with Denmark having the smallest number of gamblers with problematic behaviour. The variation might be due to national differences in gambling preferences, access to games, public policies concerning gambling, etc., but investigation of this would require further comparative research. PMID- 19541760 TI - Mortality patterns in geographical areas with a high vs. low Sami population density in Arctic Norway. AB - AIMS: The aim of this study was to study mortality patterns in the population in different geographical locations in northern Norway on the basis of Sami population density. METHODS: Mortality statistics by sex and age from 1991 to 2006 were obtained from Statistics Norway. Mortality rates, including infant mortality rates, were compared across geographical locations. The data material was divided into within and outside the the geographical areas of the Sami Development Fund (SUF) and into coastal and inland residence. Differences in mortality rates were tested by chi-quadrate tests. RESULTS: Overall, no pronounced difference in mortality rate was found in the population between geographical areas. For men, mortality rates differed only between geographical residence in the time intervals 1991-95 and 1996-2000 for the age groups 45-59 years and 60-74 years, respectively, with outside SUF area, inland having the lowest rate. For women, there were no differences between geographical areas except for the age group 60-74 years in the time period 1991-95, where SUF coast had the highest mortality rate. For the SUF area, men had the lowest probabilities for surviving the age of 74 years and women had among the highest. No geographical difference was found in infant mortality. During the time period 1991-2006, the outside SUF area, inland had an average infant mortality rate of 1.8/1000 per year, and for SUF coast it was 6.2/1000 per year. This difference was, however, not significant (p = 0.08). CONCLUSIONS: Overall, mortality rates were similar across geographical areas with low and with high Sami population density. However, indications of geographical differences in infant mortality should be investigated further. PMID- 19541761 TI - Socioeconomic inequalities in the employment impact of ischaemic heart disease: a longitudinal record linkage study in Sweden. AB - AIMS: Low socioeconomic status is associated with increased risk of ischaemic heart disease (IHD) in many European and North American countries. We hypothesize that the adverse impact of having IHD may also be heavier in lower socioeconomic groups and may constitute a mechanism for generating or reinforcing social inequalities in health. METHODS: Population registers of the 1.8 million residents of Stockholm County were used to assess the employment consequences over five years of having a diagnosis of IHD (600 cases) requiring hospital admission in 1996. We calculated annual age-standardized employment rates and age adjusted odds of leaving employment during 1997-2001 by social class for in patients and the general population. RESULTS: Men and women who had an in-patient episode for IHD had lower age-standardized employment rates than the general population. Following hospital admission, the likelihood of patients with IHD leaving employment increased annually, and by 2001 their adjusted odds were almost four times greater (odds ratio 3.95, 95% confidence interval 3.23-4.83) than for the general population. The impact of IHD on employment was more severe with decreasing social class and patients employed in low-skilled manual occupations were significantly more likely to lose employment than professional workers with the same diagnosis. Within each social class patients had significantly higher odds of leaving employment than the general population. CONCLUSIONS: In spite of its progressive employment and rehabilitation policies, our study revealed considerable and socially differentiated employment consequences of IHD in Sweden. More account needs to be taken of such differential impact in health and social policy development. PMID- 19541762 TI - Review article: chronobiology: influence of circadian rhythms on the therapy of severe pain. AB - Modern pain therapy widely follows the WHO (World Health Organization) guidelines using a three-step 'ladder' for pain relief. This escalating step scheme includes the administration in the order nonopioids, mild opioids and strong opioids, and adjuvants at any step. Analgesics should be given 'by the clock' rather than 'on demand'. However, the chronobiological parameters circadian pain rhythm, circadian efficacy of analgesics, and individual circadian need for analgesics are to be considered. The results of a multitude of studies in chronobiology are not consistent. Therefore, further studies with standardized protocols are needed that allow to assign more consistent rhythms to diseases, pain causes, and analgesic efficacy of opioids. In many cases, each patient perceives pain and its intensity individually during the time of day. By administration of analgesics over a constant or continuous dosage time fluctuations in pain perception and the outcomes of many studies in chronobiology are ignored that prove the influence of biological rhythms on the pharmacokinetic and pharmacodynamic aspects of analgesics. As different types of pain show different rhythms (highest pain intensities arising at different times of the day) analgesics should be dosed flexibly. It is also very important that drug therapy can be adjusted individually to the pain rhythm of the patient as well as to the type and cause of pain. In severe pain, therapy should be particularly careful. A flexible dosage depending on pain intensity and rapid dose adjustment are essentials of a modern pain therapy. Therefore, opioids that are flexible to use are better suited to treat the individual pain of the patient than rigid modified release oral or transdermal systems. PMID- 19541763 TI - Oncology pharmacy practice in a teaching hospital in Nepal. AB - OBJECTIVE: To highlight the patient care activities performed by pharmacists during their ward rounds in medical oncology ward of a tertiary care hospital in western region of Nepal. METHODS: The study was conducted for 3 months. Two pharmacists joined the clinicians and nurses in the ward rounds every morning as a member of healthcare team. The data used in this study was obtained from different documented files in hospital and were analyzed as per study objectives. RESULTS: During 3 months (May 20, 2008 to August 20, 2008), pharmacists provided answers to eight queries asked by clinicians and nurses for the patient care purpose. During the same period pharmacists detected four cases of medication errors and provided suggestions on them. A total of 30 adverse drug reaction (ADR) reports were reported to the regional pharmacovigilance center during the study time. Altogether there were 84 types of adverse drug reaction seen on those patients. Most of them were related to hematological system. CONCLUSION: The study evaluated the drug information provided by pharmacists, spontaneous reporting of ADRs by the pharmacists, and their intervention on treatment plan of patients during ward round. This suggests pharmacists can play a significant role on patient care when he or she joins round with other healthcare personnel in the oncology ward. PMID- 19541764 TI - Use of combined androgen blockade for advanced prostate cancer in British Columbia. AB - OBJECTIVES: Initial androgen deprivation therapy (ADT) for metastatic prostate cancer with combined androgen blockade (luteinizing-hormone releasing hormone agonist [LHRH agonist] plus antiandrogen) is not recommended in British Columbia (BC). However, this is difficult to monitor since ADT includes concurrent antiandrogen for the first month of LHRH agonist to prevent disease flare. We describe the prevalence of CAB use in BC and its financial impact. METHODS: This was a population-based, retrospective analysis. Patients started on LHRH agonist in January 2005 to December 2006 were identified from the BC Cancer Agency database. CAB was defined as greater than 1 month of antiandrogen concurrently with LHRH agonist. Incremental cost of CAB was based on an average 18 months of therapy from the pivotal CAB study. Incremental cost-effectiveness ratio (ICER) was based on life-year gained (LYG) from the Prostate Cancer Trialists' Collaborative Group meta-analysis. Estimated financial impact for 2007-2008 was based on an annual increase by 5.5% in prevalence of prostate cancer in BC. RESULTS: A total of 2751 patients were identified. CAB was used in 607 patients (22%), associated with an incremental cost of CDN$1768 and ICER of CDN$11,220/LYG per patient. Total incremental cost was CDN$1,073,176 and estimated to be CDN$1,398,644 for January 2007 to December 2008. CONCLUSION: Nearly one-quarter of patients were treated with CAB for metastatic prostate cancer even though it was not recommended in BC. Additional cost of CAB use was considerable, at CDN$1768 per patient. With increased prevalence of prostate cancer, this has important budget implication for funding agencies which do to recommend CAB. PMID- 19541765 TI - The notch target gene HES1 regulates cell cycle inhibitor expression in the developing pituitary. AB - The pituitary is an endocrine gland responsible for the release of hormones, which regulate growth, metabolism, and reproduction. Diseases such as hypopituitarism or pituitary adenomas are able to disrupt pituitary function leading to suboptimal function of the entire endocrine system. Growth of the pituitary during development and adulthood is a tightly regulated process. Hairy and enhancer of split (HES1), a transcription factor whose expression is initiated by the Notch signaling pathway, is a repressor of cell cycle inhibitors. We hypothesize that with the loss of Hes1, pituitary progenitors are no longer maintained in a proliferative state, choosing instead to exit the cell cycle. To test this hypothesis, we examined the expression of cell cycle regulators in wild-type and Hes1-deficient pituitaries. Our studies indicate that in early pituitary development [embryonic day (e) 10.5], cells contained in the Rathke's pouch of Hes1 mutants have decreased proliferation, indicated by changes in phosphohistone H3 expression. Furthermore, pituitaries lacking Hes1 have increased cell cycle exit, shown by significant increases in the cyclin-dependent kinase inhibitors, p27 and p57, from e10.5 to e14.5. Additionally, Hes1 mutant pituitaries have ectopic expression of p21 in Rathke's pouch progenitors, an area coincident with increased cell death. These observations taken together indicate a role for HES1 in the control of cell cycle exit and in mediating the balance between proliferation and differentiation, allowing for the properly timed emergence of hormone secreting cell types. PMID- 19541767 TI - Autophagy-mediated insulin receptor down-regulation contributes to endoplasmic reticulum stress-induced insulin resistance. AB - Endoplasmic reticulum (ER) stress is associated with obesity-induced insulin resistance, yet the underlying mechanisms remain to be fully elucidated. Here we show that ER stress-induced insulin receptor (IR) down-regulation may play a critical role in obesity-induced insulin resistance. The expression levels of IR are negatively associated with the ER stress marker C/EBP homologous protein (CHOP) in insulin target tissues of db/db mice and mice fed a high-fat diet. Significant IR down-regulation was also observed in fat tissue of obese human subjects and in 3T3-L1 adipocytes treated with ER stress inducers. ER stress had little effect on IR tyrosine phosphorylation per se but greatly reduced IR downstream signaling. The ER stress-induced reduction in IR cellular levels was greatly alleviated by the autophagy inhibitor 3-methyladenine but not by the proteasome inhibitor N-benzoyloxycarbonyl (Z)-Leu-Leu-leucinal (MG132). Inhibition of autophagy prevented IR degradation but did not rescue IR downstream signaling, consistent with an adaptive role of autophagy in response to ER stress induced insulin resistance. Finally, chemical chaperone treatment protects cells from ER stress-induced IR degradation in vitro and obesity-induced down regulation of IR and insulin action in vivo. Our results uncover a new mechanism underlying obesity-induced insulin resistance and shed light on potential targets for the prevention and treatment of obesity-induced insulin resistance and type 2 diabetes. PMID- 19541766 TI - Male-biased effects of gonadotropin-releasing hormone neuron-specific deletion of the phosphoinositide 3-kinase regulatory subunit p85alpha on the reproductive axis. AB - GnRH neurosecretion is subject to regulation by insulin, IGF-I, leptin, and other neuroendocrine modulators whose effects may be conveyed by activation of phosphoinositide 3-kinase (PI3K)-mediated pathways. It is not known, however, whether any of these regulatory actions are exerted directly, via activation of PI3K in GnRH neurons, or whether they are primarily conveyed via effects on afferent circuitries governing GnRH neurosecretion. To investigate the role of PI3K signaling in GnRH neurons, we used conditional gene targeting to ablate expression of the major PI3K regulatory subunit, p85alpha, in GnRH neurons. Combined in situ hybridization and immunohistochemistry confirmed reduction of p85alpha mRNA expression in GnRH neurons of GnRH-p85alpha knockout (KO) animals. Females of both genotypes exhibited estrous cyclicity and had comparable serum LH, estradiol-17beta, and FSH levels. In male GnRH-p85alphaKO mice, serum LH, testosterone, and sperm counts were significantly reduced compared with wild type. To investigate the role of the other major regulatory subunit, p85beta, on the direct control of GnRH neuronal function, we generated mice with a GnRH neuron-specific p85alpha deletion on a global betaKO background. No additional reproductive effects in male or female mice were found, suggesting that p85beta does not substitute p85 activity toward PI3K function in GnRH neurons. Our results suggest that p85alpha, and thus PI3K activity, participates in the control of GnRH neuronal activity in male mice. The sex-specific phenotype in these mice raises the possibility that PI3K activation during early development may establish sex differences in GnRH neuronal function. PMID- 19541768 TI - Identification of the HIT-45 protein from Trypanosoma brucei as an FHIT protein/dinucleoside triphosphatase: substrate specificity studies on the recombinant and endogenous proteins. AB - A new member of the FHIT protein family, designated HIT-45, has been identified in the African trypanosome Trypanosoma brucei. Recombinant HIT-45 proteins were purified from trypanosomal and bacterial protein expression systems and analyzed for substrate specificity using various dinucleoside polyphosphates, including those that contain the 5'-mRNA cap, i.e., m(7)GMP. This enzyme exhibited typical dinucleoside triphosphatase activity (EC 3.6.1.29), having its highest specificity for diadenosine triphosphate (ApppA). However, the trypanosome enzyme contains a unique amino-terminal extension, and hydrolysis of cap dinucleotides with monomethylated guanosine or dimethylated guanosine always yielded m(7)GMP (or m(2,7)GMP) as one of the reaction products. Interestingly, m(7)Gpppm(3)(N6, N6, 2'O)A was preferred among the methylated substrates. This hypermethylated dinucleotide is unique to trypanosomes and may be an intermediate in the decay of cap 4, i.e., m(7)Gpppm(3)(N6, N6, 2'O)Apm(2'O)Apm(2'O)Cpm(2)(N3, 2'O)U, that occurs in these organisms. PMID- 19541769 TI - IGF2BP1 enhances HCV IRES-mediated translation initiation via the 3'UTR. AB - The positive-strand RNA genome of the Hepatitis C virus (HCV) contains an internal ribosome entry site (IRES) in the 5'untranslated region (5'UTR) and structured sequence elements within the 3'UTR, but no poly(A) tail. Employing a limited set of initiation factors, the HCV IRES coordinates the 5'cap-independent assembly of the 43S pre-initiation complex at an internal initiation codon located in the IRES sequence. We have established a Huh7 cell-derived in vitro translation system that shows a 3'UTR-dependent enhancement of 43S pre-initiation complex formation at the HCV IRES. Through the use of tobramycin (Tob)-aptamer affinity chromatography, we identified the Insulin-like growth factor-II mRNA binding protein 1 (IGF2BP1) as a factor that interacts with both, the HCV 5'UTR and 3'UTR. We report that IGF2BP1 specifically enhances translation at the HCV IRES, but it does not affect 5'cap-dependent translation. RNA interference against IGF2BP1 in HCV replicon RNA-containing Huh7 cells reduces HCV IRES mediated translation, whereas replication remains unaffected. Interestingly, we found that endogenous IGF2BP1 specifically co-immunoprecipitates with HCV replicon RNA, the ribosomal 40S subunit, and eIF3. Furthermore eIF3 comigrates with IGF2BP1 in 80S ribosomal complexes when a reporter mRNA bearing both the HCV 5'UTR and HCV 3'UTR is translated. Our data suggest that IGF2BP1, by binding to the HCV 5'UTR and/or HCV 3'UTR, recruits eIF3 and enhances HCV IRES-mediated translation. PMID- 19541770 TI - The alpha-galactomannan Davanat binds galectin-1 at a site different from the conventional galectin carbohydrate binding domain. AB - Galectins are a sub-family of lectins, defined by their highly conserved beta sandwich structures and ability to bind to beta-galactosides, like Gal beta1-4 Glc (lactose). Here, we used (15)N-(1)H HSQC and pulse field gradient (PFG) NMR spectroscopy to demonstrate that galectin-1 (gal-1) binds to the relatively large galactomannan Davanat, whose backbone is composed of beta1-4-linked d mannopyranosyl units to which single d-galactopyranosyl residues are periodically attached via alpha1-6 linkage (weight-average MW of 59 kDa). The Davanat binding domain covers a relatively large area on the surface of gal-1 that runs across the dimer interface primarily on that side of the protein opposite to the lactose binding site. Our data show that gal-1 binds Davanat with an apparent equilibrium dissociation constant (K(d)) of 10 x 10(-6) M, compared to 260 x 10(-6) M for lactose, and a stiochiometry of about 3 to 6 gal-1 molecules per Davanat molecule. Mannan also interacts at the same galactomannan binding domain on gal 1, but with at least 10-fold lower avidity, supporting the role of galactose units in Davanat for relatively strong binding to gal-1. We also found that the beta-galactoside binding domain remains accessible in the gal-1/Davanat complex, as lactose can still bind with no apparent loss in affinity. In addition, gal-1 binding to Davanat also modifies the supermolecular structure of the galactomannan and appears to reduce its hydrodynamic radius and disrupt inter glycan interactions thereby reducing glycan-mediated solution viscosity. Overall, our findings contribute to understanding gal-1-carbohydrate interactions and provide insight into gal-1 function with potentially significant biological consequences. PMID- 19541771 TI - Unusual accumulation of sulfated glycosphingolipids in colon cancer cells. AB - The structures of glycosphingolipids from highly purified colorectal cancer cells and normal colorectal epithelial cells of 16 patients have been analyzed in fine detail (Misonou Y, Shida K, Korekane H, Seki Y, Noura S, Ohue M, Miyamoto Y. 2009. Comprehensive Clinico-Glycomic Study of 16 Colorectal Cancer Specimens: Elucidation of aberrant glycosylation and ts mechanistic causes in colorectal cancer cells. J Proteome Res. 8:2990-3005). Further structural analyses demonstrated that colon cancer cells from two patients accumulated unusual glycosphingolipids which were not observed in either colorectal cancer cells or normal colorectal epithelial cells from the other patients. Mass spectrometry analyses revealed that the unusual structures include sulfated oligosaccharides. The structures of the glycosphingolipids of the cancer cells from these two cases were analyzed by methods which include enzymatic release of carbohydrate moieties, fluorescent labeling with aminopyridine and identification using two dimensional mapping, enzymatic digestion and mass spectrometry together with methanolysis, and the use of newly synthesized sulfo-fucosylated oligosaccharides as standards. The colon cancer cells from one of the patients demonstrate a variety of oligosaccharides as major components which are sulfated at the C6 position of subterminal GlcNAc and at C3 positions of terminal galactose with or without sialylation or fucosylation. These include 6-sulfo Le(x), 6'-sialyl 6 sulfo lactosamine, and 3'-sialyl 6-sulfo Le(x), in addition to sialylated or fucosylated derivatives of type-1 and type-2 hybrid oligosaccharides. The colon cancer cells from the other patient have two kinds of sulfated oligosaccharides, a 6-sulfo Le(x) structure and a 3'-sulfo Le(x) structure, as minor components. Taking into consideration the clinical features of the two patients, the biological significance of sulfated glycosphingolipids on cancer cells is discussed. PMID- 19541772 TI - Screening for elevated levels of fear-avoidance beliefs regarding work or physical activities in people receiving outpatient therapy. AB - BACKGROUND: Screening people for elevated levels of fear-avoidance beliefs is uncommon, but elevated levels of fear could worsen outcomes. Developing short screening tools might reduce the data collection burden and facilitate screening, which could prompt further testing or management strategy modifications to improve outcomes. OBJECTIVE: The purpose of this study was to develop efficient yet accurate screening methods for identifying elevated levels of fear-avoidance beliefs regarding work or physical activities in people receiving outpatient rehabilitation. DESIGN: A secondary analysis of data collected prospectively from people with a variety of common neuromusculoskeletal diagnoses was conducted. METHODS: Intake Fear-Avoidance Beliefs Questionnaire (FABQ) data were collected from 17,804 people who had common neuromusculoskeletal conditions and were receiving outpatient rehabilitation in 121 clinics in 26 states (in the United States). Item response theory (IRT) methods were used to analyze the FABQ data, with particular emphasis on differential item functioning among clinically logical groups of subjects, and to identify screening items. The accuracy of screening items for identifying subjects with elevated levels of fear was assessed with receiver operating characteristic analyses. RESULTS: Three items for fear of physical activities and 10 items for fear of work activities represented unidimensional scales with adequate IRT model fit. Differential item functioning was negligible for variables known to affect functional status outcomes: sex, age, symptom acuity, surgical history, pain intensity, condition severity, and impairment. Items that provided maximum information at the median for the FABQ scales were selected as screening items to dichotomize subjects by high versus low levels of fear. The accuracy of the screening items was supported for both scales. LIMITATIONS: This study represents a retrospective analysis, which should be replicated using prospective designs. Future prospective studies should assess the reliability and validity of using one FABQ item to screen people for high levels of fear-avoidance beliefs. CONCLUSIONS: The lack of differential item functioning in the FABQ scales in the sample tested in this study suggested that FABQ screening could be useful in routine clinical practice and allowed the development of single-item screening for fear-avoidance beliefs that accurately identified subjects with elevated levels of fear. Because screening was accurate and efficient, single IRT-based FABQ screening items are recommended to facilitate improved evaluation and care of heterogeneous populations of people receiving outpatient rehabilitation. PMID- 19541773 TI - A controlled examination of medical and psychosocial factors associated with low back pain in combination with widespread musculoskeletal pain. AB - BACKGROUND: Little is known about chronic low back pain (CLBP) in combination with widespread musculoskeletal pain (WMP). OBJECTIVE: This study examined factors that may be common to these conditions, with the objective of detecting factors that could improve the course of these diseases. DESIGN: This was a prospective case control study. PATIENTS AND INTERVENTION: A group of patients with CLBP and WMP (CLBP+WMP group, n=97) was compared with a group of individuals who were pain-free and without a history of musculoskeletal problems (control group, n=97) and with a group of patients with CLBP but without WMP (CLBP-only group, n=52). The mean age of the participants was 42.9 years (SD=8.74); 76% were women, and 24% were men. MEASUREMENTS: A total of 74 variables were measured, including sociodemographic, physical, and psychosocial variables. After univariate examination for group differences and analyses of variables available for all 3 groups, logistic regression on selected factors was performed. The alpha level was set at .05, but was adjusted to avoid randomly significant results. RESULTS: For a number of variables, significant differences among the 3 groups were observed. For regression model 1 for the CLBP+WMP and control groups, 4 out of 9 variables showed significant likelihood tests: income (lower in the WMP group), depression, anxiety, and fear-avoidance behavior. For regression model 2, 2 out of 13 variables showed significant likelihood tests: endurance capacity (more in the CLBP+WMP group) and balance capability (worse in the CLBP+WMP group). The models predicted at least 91.2% of all cases to the correct group. The regression analysis regarding the CLBP+WMP and CLBP-only groups predicted 86.7% of all cases to the correct group. Three out of 10 variables showed significant likelihood tests: high disability, fear-avoidance behavior, and number of treatments. LIMITATIONS: Some variables in testing the patients with WMP and the individuals who were pain-free were not used with the patients with CLBP only. CONCLUSIONS: Patients with CLBP and WMP should be examined for indicated physical and psychosocial factors. Therapeutic management should consider them in the early stage of the disease. These findings also might apply to patients with fibromyalgia or myofascial pain. PMID- 19541774 TI - Comparison of maximum tolerated muscle torques produced by 2 pulse durations. AB - BACKGROUND: Neuromuscular electrical stimulation (NMES) is an effective therapeutic technique for strengthening weak muscles. A positive dose-response relationship exists between the elicited muscle forces during training and strength (force-generating capacity) gains. Patient discomfort limits NMES muscle forces, potentially compromising efficacy. OBJECTIVE: The purpose of this study was to compare the NMES muscle torques produced by stimulation trains consisting of 2 different pulse durations. DESIGN: During a single testing session, the 2 pulse duration conditions (50 and 200 microseconds) were tested on the opposite lower extremities of the participants. METHODS: The study participants were 10 adults without remarkable medical histories. The maximum tolerated isometric knee extensor torque was the primary dependent variable. The peak currents and phase charges that produced the maximally tolerated torques, as well as the sensory, motor, and pain thresholds for the 2 pulse conditions, were compared. RESULTS: The 200-microsecond pulse duration condition resulted in participants tolerating significantly greater muscle torques; it was associated with significantly greater phase charges but significantly lower peak currents. LIMITATIONS: This study only compared muscle torques in response to stimulation trains consisting of pulses with short (50-microsecond) or medium (200-microsecond) durations and did not examine long ( approximately 400- to 600-microsecond) durations. Furthermore, the result of this study may not apply to NMES that uses stimulation patterns other than monophasic, square-wave pulsed current. CONCLUSIONS: It has been suggested that short pulse durations are most appropriate for NMES because they are less likely to recruit nociceptors. The results of this study, however, support the use of a medium pulse duration rather than a short pulse duration when the goal is to produce a maximum torque response from a muscle. These observations may be related to the currents and phase charges for the pain thresholds for the 2 pulse duration conditions. PMID- 19541775 TI - Fractional anisotropy--threshold dependence in tract-based diffusion tensor analysis: evaluation of the uncinate fasciculus in Alzheimer disease. AB - BACKGROUND AND PURPOSE: Tract-based analysis can be used to investigate required tracts extracted from other fiber tracts. However, the fractional anisotropy (FA) threshold influences tractography analysis. The current study evaluated the influence of the FA threshold in measuring diffusion tensor parameters for tract based analysis of the uncinate fasciculus in subjects with Alzheimer disease (AD). MATERIALS AND METHODS: Subjects included 30 patients with AD and 10 healthy controls. We acquired tractographies of the uncinate fasciculus by using different FA thresholds. We measured mean FA and the apparent diffusion coefficient (ADC) along the uncinate fasciculus for different FA thresholds and evaluated the correlation between diffusion tensor parameters (FA, ADC) and the Mini-Mental State Examination (MMSE) scores. RESULTS: The uncinate fasciculus showed lower mean FA and higher mean ADC values in cases with more severe AD. A higher FA threshold led to a lower mean ADC value and a higher mean FA value along the uncinate fasciculus, whereas the relative order of measured values according to the severity of AD was not influenced by the FA threshold. An FA threshold of 0.2 showed higher correlation between mean ADC values and MMSE scores. FA thresholds of 0.15 and 0.20 showed higher correlation between mean FA values and MMSE scores. CONCLUSIONS: Appropriate selection of the FA threshold leads to higher correlation between diffusion tensor parameters and the severity of AD. For tract-based analysis of degenerative diseases such as AD, appropriate selection of the FA threshold for tractography is important. PMID- 19541776 TI - CT assessment of woodworkers' nasal adenocarcinomas confirms the origin in the olfactory cleft. AB - BACKGROUND AND PURPOSE: Endoscopic endonasal surgery let us observe that woodworkers' nasal adenocarcinomas originate in the olfactory cleft. Our aim was the identification of CT imaging features that corroborate the olfactory cleft as the site of origin for woodworkers' adenocarcinoma. MATERIALS AND METHODS: We designed a retrospective study to compare CT scans of 27 unilateral olfactory cleft adenocarcinomas with 30 cases of nasosinusal polyposis (NSP) and 33 healthy sinus controls. Enlargement of the olfactory cleft, lateralization of the ethmoidal turbinate wall, and contralateral bulging of the nasal septum were measured on coronal scans passing through crista galli and posterior half of both ocular globes. Comparisons have been performed by using analysis of variance and the Bonferroni procedure. RESULTS: The nasal septum was significantly bulging across the midline in adenocarcinoma (4.6 +/- 3 mm; range, -0.1-13.7 mm) compared with NSP (0.7 +/- 1 mm; range, -2.1-2.3 mm) or healthy sinus controls (0.5 +/- 1 mm; range, -1.2-2 mm) (P < .001). The olfactory cleft was significantly wider in adenocarcinoma (15.1 +/- 4.5 mm; range, 8.6-25.7 mm) than in NSP (3.6 +/- 0.4 mm; range, 2.8-4.6 mm) or healthy sinus controls (3.3 +/- 0.7 mm; range, 1.4-4.6 mm). The ethmoidal labyrinth width was significantly smaller on the pathologic side in adenocarcinoma (7.2 +/- 2.7 mm; range, 3.2-14.2 mm) than in the control groups (P < .001). Whereas the angle between the conchal lamina and vertical midline was close to zero degrees in NSP (0.03 +/- 2.25 degrees ; range, -5 degrees -3 degrees ) and healthy sinus controls (0.45 +/- 2.13 degrees , range, -5 degrees 5 degrees ), it reached 39.76 +/- 13.83 degrees (P < .001) in adenocarcinoma. CONCLUSIONS: Radiologists should suspect nasal adenocarcinoma on sinus CT scans showing a unilateral expanding opacity of the olfactory cavity. PMID- 19541777 TI - Cerebellar lesions in multiple system atrophy: postmortem MR imaging-pathologic correlations. AB - BACKGROUND AND PURPOSE: Cerebellar atrophy and white matter T2-hyperintensities have been characterized as cerebellar lesions of multiple system atrophy (MSA). The aim of the study was to correlate MR images with histologic findings in cerebellar lesions of MSA. MATERIALS AND METHODS: Postmortem T2-weighted images using 1.5T were compared with histologic findings in 7 postmortem-proved cases with MSA. The MR imaging findings in the cerebellar cortices and deep white matter dentate nucleus regions were compared with their histologic findings in each case. RESULTS: We detected 3 types of cerebellar changes: type 1, no apparent atrophy or signal-intensity changes; type 2, cerebellar atrophy and inhomogeneous (patchy and/or confluent) cerebellar white matter hyperintensities; and type 3, cerebellar atrophy and diffuse white matter hyperintensities. Hypointensities were seen in the dentate nucleus regions. Atrophy of the cerebellar white matter was more severe than that of cerebellar cortices, and this anatomy was well depicted on coronal images. Histologically, degeneration was more severe in the cerebellar white matter than in the cerebellar cortices. Hyperintensities in the cerebellar white matter showed loss of myelinated fibers and gliosis. Hypointensities in the dentate nucleus regions revealed diffuse ferritin deposition in preserved dentate nuclei and white matter both around and within the nuclei. CONCLUSIONS: Hyperintensities in the cerebellar white matter reflect degenerated white matter associated with loss of myelinated fibers and gliosis, whereas hypointensities in the dentate nucleus regions reflect diffuse ferritin deposition in preserved dentate nuclei and white matter around and within the nuclei. Degeneration is more severe in the cerebellar white matter than in the cerebellar cortices. PMID- 19541778 TI - Prevalence of hippocampal malrotation in a population without seizures. AB - BACKGROUND AND PURPOSE: Hippocampal malrotation (HIMAL) is a failure of hippocampal inversion that occurs during normal fetal development and has been seen on MR imaging examinations of people with epilepsy, but it has not been studied in patients without epilepsy. We intended to evaluate the prevalence of HIMAL in MR imaging examinations of patients without seizures to better understand the significance of HIMAL in the population with seizure. MATERIALS AND METHODS: A total of 497 MR imaging examinations with thin-section imaging through the temporal lobes of patients referred for conditions other than seizures were reviewed. The examinations were performed on 1.5T magnets. Sagittal T1-weighted and coronal T2-weighted images were used to evaluate each MR image for the distinctive features of HIMAL. As previously described in the literature, the criteria for HIMAL include unilateral involvement and incomplete rotation of a hippocampus that is normal in size and signal intensity but abnormally rounded in shape, with blurred inner structure. In addition, ipsilateral findings of an atypical collateral sulcus angle and atypical position and size of the fornix were noted. The corpus callosum is normal, and the temporal lobe remains normal in size, though the temporal horn may appear enlarged. RESULTS: None of the patients' examinations fulfilled all of the HIMAL criteria. Six studies satisfied 2 or more criteria, which included an abnormally rounded hippocampus and a vertical collateral sulcus. These HIMAL findings were all seen on the left. Forniceal asymmetry was the most prevalent abnormality, with 289 patients manifesting a low position of 1 fornix. CONCLUSIONS: Hippocampal malrotation is a rare finding in patients without seizures. HIMAL is therefore likely to be a pathologic finding. PMID- 19541779 TI - In utero eye development documented by fetal MR imaging. AB - BACKGROUND AND PURPOSE: To date, very limited attention has been given to ocular abnormalities or growth parameters detected by fetal MR imaging. Our objective was to retrospectively determine the relationship between different parameters of eye development and estimated gestational age in the human fetus by use of fetal MR imaging. MATERIALS AND METHODS: A retrospective study was performed to measure the transverse diameter, interocular distance, and lens diameter of the globes of 127 fetuses who had a morphologically normal central nervous system. Multiple single-shot T2 fast spin-echo images were obtained with a 1.5T magnet by use of contiguous 3-mm intervals in at least 2 orthogonal planes. Loess curves were fitted to explore the relationship between gestational age and each of the 3 measurements of interest. Different models were compared statistically to determine the model of best fit. RESULTS: For each variable of interest, the "best" model of eye growth was a quadratic function. Specifically, lens growth seems to plateau after 36 weeks of gestation, interocular distance plateaus after 36 weeks of gestation, and globe growth plateaus after 42 weeks of gestation. CONCLUSIONS: The lens, orbit, and interocular distance growth of the fetus can be demonstrated on fetal MR imaging. All 3 measurements suggest a quadratic model of growth, which indicates slowing of growth toward the end of gestation. PMID- 19541780 TI - ScreenTape as a tool for the rapid differentiation of Mycobacterium tuberculosis isolates. PMID- 19541781 TI - Penicillin/ampicillin efficacy among children with severe pneumonia due to penicillin-resistant pneumococcus (MIC=4 microg ml(-1)). PMID- 19541782 TI - Characterization of cytotoxic necrotizing factor 1-producing Escherichia coli strains from faeces of healthy macaques. AB - Twenty-five (27 %) of 92 clinically normal macaques were found to have beta haemolytic Escherichia coli isolated from their faeces. Five of six isolates chosen for further characterization had multiple antibiotic resistance and were PCR-positive for cytotoxic necrotizing factor 1 (cnf1) with a demonstrated cytopathic effect in vitro. By repetitive element sequence-based PCR genotyping, genetic similarity was established for selected isolates. We believe this to be the first report of E. coli strains producing CNF1 in non-human primates. PMID- 19541783 TI - Levofloxacin-resistant-Streptococcus mitis endophthalmitis: a unique presentation of bacterial endocarditis. AB - Endogenous endophthalmitis is a rare complication of infective endocarditis and has been decreasing due to the availability of effective antibiotics. We highlight a case of endogenous endophthalmitis due to levofloxacin-resistant Streptococcus mitis presenting as infective endocarditis. Endogenous endophthalmitis should be considered as a manifestation of an underlying systemic disease, especially in patients who present with non-specific signs and symptoms with no obvious source of precipitating infection. PMID- 19541784 TI - High prevalence of bla(CTX-M-1)-group extended-spectrum beta-lactamase genes in Enterobacteriaceae isolates from Emilia-Romagna, Italy. PMID- 19541785 TI - Plasmid-mediated quinolone resistance in Salmonella from South Africa. PMID- 19541786 TI - Vertebral osteomyelitis and discitis due to Gardnerella vaginalis. AB - Gardnerella vaginalis is a facultative anaerobic Gram-variable pleomorphic rod that forms part of the normal vaginal flora. It is most commonly associated with infection of the genital tract in women, but recognition of extravaginal G. vaginalis infection is becoming more frequent. We describe an unusual case of G. vaginalis vertebral osteomyelitis and discitis in a 38-year-old woman with no apparent predisposing factors. PMID- 19541787 TI - A newly identified protein of Leptospira interrogans mediates binding to laminin. AB - Pathogenic Leptospira is the aetiological agent of leptospirosis, a life threatening disease that affects populations worldwide. The search for novel antigens that could be relevant in host-pathogen interactions is being pursued. These antigens have the potential to elicit several activities, including adhesion. This study focused on a hypothetical predicted lipoprotein of Leptospira, encoded by the gene LIC12895, thought to mediate attachment to extracellular matrix (ECM) components. The gene was cloned and expressed in Escherichia coli BL21 Star (DE3)pLys by using the expression vector pAE. The recombinant protein tagged with N-terminal hexahistidine was purified by metal charged chromatography and characterized by circular dichroism spectroscopy. The capacity of the protein to mediate attachment to ECM components was evaluated by binding assays. The leptospiral protein encoded by LIC12895, named Lsa27 (leptospiral surface adhesin, 27 kDa), bound strongly to laminin in a dose dependent and saturable fashion. Moreover, Lsa27 was recognized by antibodies from serum samples of confirmed leptospirosis specimens in both the initial and the convalescent phases of the disease. Lsa27 is most likely a surface protein of Leptospira as revealed in liquid-phase immunofluorescence assays with living organisms. Taken together, these data indicate that this newly identified membrane protein is expressed during natural infection and may play a role in mediating adhesion of L. interrogans to its host. PMID- 19541788 TI - N-Chlorotaurine shows high in vitro activity against promastigotes and amastigotes of Leishmania species. AB - Protozoan parasites of the genus Leishmania are the causative agents of life threatening visceral as well as cutaneous and mucocutaneous leishmaniasis. First line drugs are antimonials, but toxicity and resistance in some endemic areas cause serious problems. In the current study, the antileishmanial activity of the weak oxidant N-chlorotaurine (NCT) was investigated. NCT is a derivative of the amino acid taurine produced by granulocytes and monocytes during oxidative burst, but can also be synthesized chemically and used topically as an antiseptic at a concentration of 1 % (55 mM) in vivo. NCT susceptibility tests were performed in vitro with promastigotes and amastigotes of Leishmania infantum and Leishmania donovani. As NH(4)Cl is known to increase the activity of NCT by the formation of monochloramine (NH(2)Cl), co-treatment assays were included in the study. Mean EC(50) values after 1 h of treatment were 5.94 mM for L. infantum and 9.8 mM for L. donovani promastigotes. Co-treatment with 5.5 mM NCT plus 19 mM NH(4)Cl led to complete killing of promastigotes of both strains within 15 min. Amastigotes were inactivated by treatment with 2 mM NCT alone. The results of this study indicate a high potential of NCT against Leishmania species. PMID- 19541789 TI - Detection of Aspergillus DNA by a nested PCR assay is able to improve the diagnosis of invasive aspergillosis in paediatric patients. AB - Fungal infections are a leading cause of morbidity and mortality in severely immunocompromised patients and have been increasing in incidence in recent years. Invasive aspergillosis (IA) is the most common filamentous fungal infection and is, in adults as well as in children, difficult to diagnose. Several PCR assays to detect Aspergillus DNA have been established, but so far, studies on molecular tools for the diagnosis of IA in children are few. We evaluated the results of a nested PCR assay to detect Aspergillus DNA in clinical samples from paediatric and adolescent patients with suspected IA. Blood and non-blood samples from immunocompromised paediatric and adolescent patients with suspected invasive fungal infection were sent for processing Aspergillus PCR to our laboratory. PCR results from consecutive patients from three university children's hospitals investigated between November 2000 and January 2007 were evaluated. Fungal infections were classified according to the EORTC classification on the grounds of clinical findings, microbiology and radio-imaging results. Two hundred and ninety-one samples from 71 patients were investigated for the presence of Aspergillus DNA by our previously described nested PCR assay. Two, 3 and 34 patients had proven, probable and possible IA, respectively. Sensitivity (calculated from proven and probable patients, n=5) and specificity (calculated from patients without IA, n=32) rates of the PCR assay were 80 and 81 %, respectively. Our nested PCR assay was able to detect Aspergillus DNA in blood, cerebrospinal fluid and bronchoalveolar lavage samples from paediatric and adolescent patients with IA with high sensitivity and specificity rates. PMID- 19541791 TI - Cytokines and angiogenic factors in patients with metastatic renal cell carcinoma treated with interferon-alpha: association of pretreatment serum levels with survival. AB - BACKGROUND: To correlate serum cytokine and angiogenic factor (CAF) levels with overall survival (OS) in metastatic renal cell carcinoma (mRCC) treated with interferon-alpha (IFN-alpha). PATIENTS AND METHODS: Serum CAF levels were measured in 103 patients treated on a randomized trial with IFN-alpha 0.5 million units (MU) twice daily or 5 MU daily. Concentrations of 17 analytes were determined by multiplex bead immunoassays [vascular endothelial growth factor A (VEGF(A)) and several cytokines] or enzyme-linked immunosorbent assay (basic fibroblast growth factor). We used proportional hazards models to evaluate the effect of CAF levels and clinical factors on OS. RESULTS: Pretreatment serum interleukin (IL) 5, IL-12 p40, VEGF(A), and IL-6 levels and Memorial Sloan Kettering Cancer Center risk grouping independently correlated with OS, with hazard ratios of 2.33, 2.00, 2.07, 1.82, and 0.39, respectively (concordance index = 0.69 for the combined model versus 0.60 for the CAF model versus 0.52 for the clinical model). Based on an index derived from these five risk factors (RFs), patients with 0-2 RF had a median OS time of 32 months versus 9 months for patients with 3-5 RF (P < 0.0001). CONCLUSIONS: Serum CAF profiling contributes to prognostic evaluation in mRCC and helps to identify a subset of patients with 20% 5-year OS. PMID- 19541792 TI - Randomized, double-blind, dose-ranging trial of the oral neurokinin-1 receptor antagonist casopitant mesylate for the prevention of cisplatin-induced nausea and vomiting. AB - BACKGROUND: Casopitant mesylate is a novel, oral neurokinin-1 receptor antagonist with demonstrated antiemetic efficacy. We conducted a randomized, double-blind, controlled phase II trial to evaluate three casopitant doses as part of a triple therapy regimen for the prevention of nausea and vomiting associated with high dose cisplatin. The aim of the study was to detect a dose response. PATIENTS AND METHODS: A total of 493 patients with solid tumors receiving a first cycle of cisplatin > or =70 mg/m(2) were randomly assigned among six treatment arms. The primary analysis compared a control arm [ondansetron/dexamethasone (Ond/Dex)] with three investigational treatments (Ond/Dex plus oral casopitant 50, 100, or 150 mg administered daily for 3 days). Two exploratory arms were included: one evaluating a single oral casopitant dose of 150 mg added to standard Ond/Dex and another with 3-day oral aprepitant-based therapy (Ond/Dex plus aprepitant 125 mg day 1, 80 mg days 2-3). RESULTS: The complete response (CR) rate (defined as no vomiting, retching, rescue therapy, or premature discontinuation) was significantly increased in each casopitant arm relative to control over the 120-h evaluation period: 76% (50 mg), 86% (100 mg), 77% (150 mg), and 60% with control (P = 0.0036). The CR rate for the single oral dose regimen was similar to the CR rate reported for the 3-day regimens. No differences were observed in the incidence of nausea or significant nausea among groups in the primary analysis. The most common adverse events related to treatment included headache (n = 10) and hiccups (n = 14). CONCLUSION: All doses of oral casopitant as a 3-day regimen (and likely as a 150-mg single oral dose) in combination with Ond/Dex provided significant improvement in the prevention of cisplatin-induced emesis. PMID- 19541793 TI - Prognostic significance of mid- and post-ABVD PET imaging in Hodgkin's lymphoma: the importance of involved-field radiotherapy. AB - BACKGROUND: Although positron emission tomography (PET) response to chemotherapy (CT) has prognostic significance in Hodgkin's lymphoma (HL), it is unclear whether patients with 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG)-PET positivity during and/or after CT can be rendered disease free with consolidative involved-field radiotherapy (IFRT). METHODS: Patients with HL treated with adriamycin, bleomycin, vinblastine and dacarbazine (ABVD)-based CT and radiotherapy (RT) at our institution from January 2000 to March 2007 were eligible. All patients had either a post-treatment PET or PET-CT before initiation of RT or a negative midtreatment PET or PET-CT. The primary end point was failure-free survival (FFS) for patients with and without residual FDG avidity after ABVD. The treatment outcome of patients with interim PET positivity during CT was also reported. RESULTS: Seventy-three patients were included in this study. Twenty patients (out of 46) were PET positive on interim PET, and 13 patients (out of 73) were PET positive at the conclusion of CT. At a median follow-up of 3.4 years for surviving patients, the 2-year FFSs for patients PET negative versus PET-positive disease after ABVD were 95% and 69%, respectively (P < 0.01). On bivariable Cox regression, post-ABVD positivity (hazard ratio 4.8, P = 0.05) was predictive of disease recurrence after controlling for bulky disease. Of the 20 patients with interim PET positivity, three recurred, with a 2-year FFS of 85%. Among the 13 patients with interim PET positivity, but became PET negative at the completion of CT, the 2-year FFS was 92%. CONCLUSION: Sixty-nine per cent of patients with residual FDG avidity after ABVD were free of disease after consolidative RT, indicating a majority of patients with persistent lymphoma can be cured by sterilizing this PET-positive disease. PMID- 19541794 TI - Fusarial toxin-induced toxicity in cultured cells and in isolated mitochondria involves PTPC-dependent activation of the mitochondrial pathway of apoptosis. AB - Mycotoxins produced by the Fusarium molds can cause a variety of human diseases and economic losses in livestock. Fusaria produce predominantly two types of mycotoxins: the nonestrogenic trichothecenes including T-2 toxin and the mycoestrogens such as zearalenone (ZEN). In a previous report, we demonstrated that the hepatotoxicity of these mycotoxins involves the mitochondrial pathway of apoptosis. Here, we observed that both fusarotoxins induced cell death by a mitochondria-dependent apoptotic process which includes opening of the mitochondrial permeability transition pore complex (PTPC), loss of mitochondrial transmembrane potential, increase in O(2)(.-) production, mitochondrial relocalization of Bax, cytochrome c release, and caspase activation. Studies performed on isolated mouse liver mitochondria showed that both ZEN and T-2 toxin might act directly on mitochondria to induce a PTPC-dependent permeabilization of mitochondrial membranes. Moreover, they may target different members of PTPC. Indeed, although the inner membrane protein adenine nucleotide translocase could be the target of T-2 toxin, ZEN seems to target the outer membrane protein voltage-dependent anion channel. Cells pretreatment with the p53 inhibitor pifithrin-alpha suggested that ZEN but not T-2 toxin triggered a p53-dependent mitochondrial apoptotic pathway. Finally, mitochondrial alterations induced by ZEN and T-2 toxin are mediated by Bcl-2 family proteins, such as Bax, and prevented by Bcl-x(L) and to a lesser extent by Bcl-2. Taken together, these data indicate that mitochondria play a pivotal role in both ZEN- and T-2 toxin-induced apoptosis and that PTPC members and proteins of Bcl-2 family should be interesting targets to overcome fusarotoxin toxicity. PMID- 19541795 TI - Atrazine oral exposure of peripubertal male rats downregulates steroidogenesis gene expression in Leydig cells. AB - In the present study, we investigated the effects of oral dosing of atrazine (2 chloro-4-ethylamino-6-isopropylamino-s-triazine) to peripubertal male rats (50 and 200 mg/kg body weight daily from postnatal days 23-50) on ex vivo Leydig cell steroidogenesis. Leydig cells from treated rats were characterised by significant decline in mRNA transcripts of several genes responsible for steroidogenesis: luteinizing hormone receptor (LHR), scavenger receptor-B1, steroidogenic acute regulatory protein, translocator protein, steroidogenic factor-1, phosphodiesterase 4B, 3beta-hydroxysteroid dehydrogenase (HSD), CYP17A1, and 17betaHSD. In the presence of human chorion gonadotropin, the dose-dependent decrease in extracellular cAMP level and accordingly strong inhibition of androgenesis were obtained. The transcription of LHR gene in Leydig cells of atrazine-treated rats was downregulated in a dose-dependent manner, which could be the reason for reduction in cAMP level and expression of cAMP-dependent genes. To clarify the activity of the steroidogenic enzymes responsible for androgenesis, purified Leydig cells were challenged with different steroid substrates (22OH-cholesterol, pregnenolone, progesterone, and Delta(4) androstenedione), and the obtained results indicated inhibition of androgen production in Leydig cells isolated from atrazine-treated animals in the presence of all those substrates. However, when Leydig cells were challenged with 22OH cholesterol, the progesterone level in the incubation medium was unchanged, indicating that decrease in cholesterol transport and/or CYP17A1 and 17betaHSD activity are most probably responsible for inhibition of androgen production after the addition of different substrates. Our results demonstrated that in vivo exposure to atrazine affects Leydig cell steroidogenesis via the inhibition of steroidogenesis gene expression, which is accompanied by decreased androgenesis. PMID- 19541796 TI - Long-term exposure to zidovudine delays cell cycle progression, induces apoptosis, and decreases telomerase activity in human hepatocytes. AB - Zidovudine (3'-azido-3'-deoxythymidine; AZT), which is currently used in the treatment of acquired immunodeficiency syndrome, has been shown to have anticancer properties. In the present study, we examined the mechanisms contributing to increased sensitivity of cancer cells to the growth-inhibitory effects of AZT. This was accomplished by incubating a hepatoma cell line (HepG2) and a normal liver cell line (THLE2) with AZT in continuous culture for up to 4 weeks and evaluating the number of viable and necrotic cells, the induction of apoptosis, cell cycle alterations, and telomerase activity. In HepG2 cells, AZT (2-100 microM) caused significant dose-dependent decreases in the number of viable cells at exposures > 24 h. During a 1-week recover period, there was only a slight increase in the number of viable cells treated with AZT. The decrease in viable cells was associated with an induction of apoptosis, a decrease in telomerase activity, and S and G2/M phase arrest of the cell cycle. During the recovery period, the extent of apoptosis and telomerase activity returned to control levels, whereas the disruption of cell cycle progression persisted. Western blot analysis indicated that AZT caused a decrease in checkpoint kinase 1 (Chk1) and kinase 2 (Chk2) and an increase in phosphorylated Chk1 (Ser345) and Chk2 (Thr68). Similar effects, to lesser extent, were observed in THLE2 cells given much higher concentrations of AZT (50-2500 microM). These data show that HepG2 cells are much more sensitive than THLE2 cells to AZT. They also indicate that a combination of a delay of cell cycle progression, an induction of apoptosis, and a decrease in telomerase activity is contributing to the decrease in the number of viable cells from AZT treatment, and that checkpoint enzymes Chk1 and Chk2 may play an important role in the delay of cell cycle progression. PMID- 19541797 TI - Nearest-neighbor and logistic regression analyses of clinical and heart rate characteristics in the early diagnosis of neonatal sepsis. AB - OBJECTIVES: To test the hypothesis that nearest-neighbor analysis adds to logistic regression in the early diagnosis of late-onset neonatal sepsis. DESIGN: The authors tested methods to make the early diagnosis of neonatal sepsis using continuous physiological monitoring of heart rate characteristics and intermittent measurements of laboratory values. First, the hypothesis that nearest-neighbor analysis makes reasonable predictions about neonatal sepsis with performance comparable to an existing logistic regression model was tested. The most parsimonious model was systematically developed by excluding the least efficacious clinical data. Second, the authors tested the hypothesis that a combined nearest-neighbor and logistic regression model gives an outcome prediction that is more plausible than either model alone. Training and test data sets of heart rate characteristics and laboratory test results over a 4-y period were used to create and test predictive models. MEASUREMENTS: Nearest-neighbor, regression, and combination models were evaluated for discrimination using receiver-operating characteristic areas and for fit using the Wald statistic. RESULTS: Both nearest-neighbor and regression models using heart rate characteristics and available laboratory test results were significantly associated with imminent sepsis, and each kind of model added independent information to the other. The best predictive strategy employed both kinds of models. CONCLUSION: The authors propose nearest-neighbor analysis in addition to regression in the early diagnosis of subacute, potentially catastrophic illnesses such as neonatal sepsis, and they recommend it as an approach to the general problem of predicting a clinical event from a multivariable data set. PMID- 19541798 TI - Gender and age differences in facial expressions. AB - The aim of this research was to determine a reliable method for quantitatively evaluating the facial expressions of children and adults in order to assess their dependence on age and gender. This study evaluated 80 healthy subjects divided into four groups: 20 girls (mean age 10.6 years), 20 boys (mean age 10.8 years), 20 females (average age 25.6 years), and 20 males (average age 27.0 years). A video was used to record each individual executing three facial expressions: a rest pose, a lip pucker, and a posed smile. Representative video frames were chosen for each individual's expressions; they were digitized and then analysed with software that extracted a set of horizontal and vertical distances of the face. All distances measured in the posed smile and lip pucker were expressed as a percentage change from the rest pose. Statistical analysis with a two-way multivariate analysis of variance (MANOVA) was performed, with gender and age as the independent variables. It was evident that the ability to produce certain facial expressions differs between groups of individuals due to gender and age. Males had a greater upward vertical movement capacity in the studied facial expressions than females. Females had a more pronounced horizontal component in the posed smile. There was a trend from childhood to adulthood showing an increase in the percentage change in most vertical movements. This trend was present in both genders, though more pronounced in males. Using a robust quantitative method for collecting and analysing facial expressions, gender differences in adults were detected as well as differences between adults and children. The trend toward increasing vertical movements in adults compared with children suggests the possibility that the mimic musculature is developmentally regulated. PMID- 19541799 TI - Molecular aspects of thyroid hormone transporters, including MCT8, MCT10, and OATPs, and the effects of genetic variation in these transporters. AB - Thyroid hormone is a pleiotropic hormone with widespread biological actions. For instance, adequate levels of thyroid hormone are critical for the development of different tissues such as the central nervous system, but are also essential for the regulation of metabolic processes throughout life. The biological activity of thyroid hormone depends not only on serum thyroid hormone levels, but is also regulated at the tissue level by the expression and activity of deiodinases, which activate thyroid hormone or mediate its degradation. In addition, thyroid hormone transporters are necessary for the uptake of thyroid hormone into target tissues. With the discovery of monocarboxylate transporter 8 (MCT8) as a specific thyroid hormone transporter and the finding that mutations in this transporter lead to a syndrome of severe psychomotor retardation and elevated serum 3,3',5 tri-iodothyronine levels known as the Allan-Herndon-Dudley syndrome, the interest in this area of research has greatly increased. In this review, we will focus on the molecular aspects of thyroid hormone transporters, including MCT8, MCT10, organic anion transporting polypeptides, and the effects of genetic variation in these transporters. PMID- 19541802 TI - Asbestos, lung cancer and mesothelioma in the British Journal of Industrial Medicine. PMID- 19541800 TI - Differential actions of estrogen and SERMs in regulation of the actin cytoskeleton of endometrial cells. AB - Estrogen and selective estrogen receptor modulators (SERMs) differentially impact endometrial cell function, however, the biological basis of these differences is not established. Deregulated cell adhesion to the extracellular matrix, cell movement and invasion are related to endometrial disorders, such as endometriosis or endometrial cancer. Remodeling of the actin cytoskeleton is required to achieve cell adhesion and movement. Estrogen receptor (ER) regulates actin and cell membrane remodeling through extra-nuclear signaling cascades. In this article, we show that administration of 17beta-estradiol (E2) and tamoxifen (TAM) to immortalized Ishikawa endometrial cells or to human endometrial stromal cells (ESC) results in remodeling of actin fibers and cell membrane. This is linked to rapid phosphorylation on Thr(558) of the actin-binding protein moesin and enhanced migration and invasion of normal and Ishikawa cells. Raloxifene (RAL) does not result in moesin activation or actin remodeling. When endometrial cells are exposed to E2 in the presence of TAM or RAL, both SERMs interfere with the recruitment of moesin, with the remodeling of the cytoskeleton, and with cell movement and migration induced by E2. The differential actions of E2, TAM and RAL are linked to a distinct modulation of the extra-nuclear signaling of ER to G proteins and to the Rho-associated kinase. These findings increase our understanding of the actions of estrogen and SERMs in endometrial cells and highlight potential molecular targets to interfere with the estrogen-related altered cell adhesion encountered in endometrial disorders. PMID- 19541803 TI - From cross-sectional survey to cohort study. PMID- 19541804 TI - Assessing workplace exposures: turning to the past for guidance. PMID- 19541805 TI - Suicide among male regular UK Armed Forces personnel, 1984-2007. AB - OBJECTIVE: To examine the number of suicide and open verdict deaths in the regular UK Armed Forces and to make comparisons with the UK general population. METHODS: Age and calendar year-adjusted standardised mortality ratios (SMRs) and 95% CI were calculated to compare the number of suicides among the UK Armed Forces with the general population of the UK. RESULTS: Between 1984 and 2007, there were 694 suicide and open verdict deaths among male UK Armed Forces personnel. The UK Armed Forces had statistically significantly fewer suicides than expected compared with the UK general population (SMR = 58, 95% CI 54 to 63, based on 694 deaths). This was evident for each of the three Services (Naval Service, Army and Royal Air Force). For each age group, the number of suicides in each Service was lower than the number expected based on UK general population rates, except for Army males under 20 years of age, where there were 1.5 times more deaths than expected (SMR = 150, 95% CI 118 to 190, based on 68 deaths). CONCLUSION: The UK Armed Forces are subject to a number of unique occupational stressors, so it is reassuring that they experience lower than expected numbers of suicides in comparison with the UK general population. This is true for each Service and all age groups except young Army males. PMID- 19541806 TI - Maternal occupational exposure to solvents and congenital malformations: a prospective study in the general population. AB - OBJECTIVE: To study the relations between maternal occupational exposure to solvents during pregnancy and the risk of congenital malformations. METHODS: A prospective population-based cohort, specifically designed to study the impact of maternal exposure to solvents on the risk of congenital malformations, began in 2002 in three districts of Brittany (France). 3421 pregnant women were recruited until the end of 2005 by physicians before 19 weeks of gestation and followed through birth. Information on pregnancy outcomes was obtained from the hospital. Occupational exposure to solvents at the beginning of pregnancy was assessed from the women's self-reported occupational exposures at inclusion and from a job exposure matrix (JEM). Sociodemographic characteristics, lifestyle factors, medical history, diseases during pregnancy were obtained at inclusion and from hospital records. Analyses were restricted to working women (n = 3005). Logistic regressions were used to adjust for potential confounders. RESULTS: 30.2% of the working women declared regular exposure to at least one product that may contain solvents. 21.3% of them were classified at least in the medium exposure category using the JEM. Occupations mainly classified as exposed by both assessment methods were hairdressers, nurses' aides, nurses and chemists/biologists. Significant associations were found between major congenital malformations and maternal occupational exposure to solvents, assessed by both self-report odds ratio (OR = 2.48, 95% CI 1.4 to 4.4 for regular exposure vs no exposure) and the JEM (OR = 3.48, 95% CI 1.4 to 8.4 for highest level of exposure vs no exposure). A significant dose-response trend was observed with both assessment methods. Several subgroups of major malformations were associated with maternal exposure to solvents (oral clefts, urinary malformations and male genital malformations). CONCLUSION: This study provides further evidence of an association between exposure to solvents during pregnancy and the risk of major malformations. PMID- 19541807 TI - Identifying tumour sites in the IARC Monographs. PMID- 19541808 TI - H-Y incompatibility predicts short-term outcomes for kidney transplant recipients. AB - A recent report suggested that female recipients of male deceased-donor kidneys are at increased risk for graft failure because of H-Y antigen mismatch. In an attempt to confirm and extend these results, we studied all adult recipients of deceased-donor kidney transplants from 1990 through 2004 in the US Renal Data System. Compared with all other gender combinations, female recipients of male donor kidneys had a 12% increased risk for graft failure at 1 yr (hazard ratio 1.12; 95% confidence interval 1.05 to 1.19) but no excess risk at 10 yr (hazard ratio 1.03; 95% confidence interval 0.98 to 1.07). We observed a similar pattern of short- and long-term risk for both death-censored graft failure and mortality. The main results were consistent across several prespecified patient subgroups and were robust to sensitivity analyses. In conclusion, compared with other recipient-donor gender combinations, female recipients of male donor kidney transplants in the United States have an increased short-term risk but not long term risk for adverse outcomes. PMID- 19541810 TI - Defining contrast-induced nephropathy. PMID- 19541811 TI - Reliability of Medicare claim forms for outcome studies in kidney transplant recipients: epidemiology in clinical outcome trials. PMID- 19541809 TI - Inhibition of integrin-linked kinase attenuates renal interstitial fibrosis. AB - Integrin-linked kinase (ILK) is an intracellular serine/threonine protein kinase that regulates cell adhesion, survival, and epithelial-to-mesenchymal transition (EMT). In this study, we investigated the kinase activity of ILK during tubular EMT induced by TGF-beta1 and examined the therapeutic potential of an ILK inhibitor in obstructive nephropathy. TGF-beta1 induced a biphasic activation of ILK in renal tubular epithelial cells, with rapid activation starting at 5 min and the second wave of activation peaking at 24 h; the latter paralleled the induction of ILK protein expression. Pharmacologic inhibition of ILK with small molecule inhibitor QLT-0267 abolished TGF-beta1-induced phosphorylation of Akt and glycogen synthase kinase-3beta, suppressed cyclin D1 expression, and largely restored the expression of E-cadherin and zonula occludens 1. Inhibition of ILK also blocked TGF-beta1-mediated induction of fibronectin, Snail1, plasminogen activator inhibitor 1, and matrix metalloproteinase 2. In a mouse model of obstructive nephropathy, administration of QLT-0267 inhibited beta-catenin accumulation; suppressed Snail1, alpha-smooth muscle actin, fibronectin, vimentin, and type I and type III collagen expression; and reduced total tissue collagen content. Inhibition of ILK did not affect kidney structure or function in normal mice. These findings suggest that increased ILK activity mediates EMT and the progression of renal fibrosis. Pharmacologic inhibition of ILK signaling may hold therapeutic potential for fibrotic kidney diseases. PMID- 19541812 TI - Intradialytic administration of daptomycin in end stage renal disease patients on hemodialysis. AB - BACKGROUND AND OBJECTIVES: Infusion of intravenous antibiotics after hemodialysis (HD) may delay initiation of treatment for the next HD shift. Intradialytic administration of drugs such as vancomycin during the final hour of HD obviates these delays. Daptomycin has potent activity against Gram-positive bacteria, but the manufacturer recommends that the dose be infused after HD ends. This study determined the pharmacokinetics of intradialytically dosed daptomycin in patients with ESRD. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This prospective crossover study compared single-dose daptomycin (6 mg/kg, 30-min intravenous infusion) pharmacokinetics administered after HD versus during the last part of HD with high-permeability (HP) and low-permeability (LP) dialyzers to seven patients who had ESRD and were on thrice-weekly HD. Serial blood samples were collected to determine daptomycin serum concentrations and protein binding. Statistical analysis was done using linear mixed model analysis. RESULTS: The maximum serum concentration observed with a 6 mg/kg post-HD dose was 61.1 +/- 7.6 microg/ml with a mean protein binding of 89.2%. Intradialytic daptomycin administration resulted in reduced maximum serum concentration and area under the curve values that were approximately 12 to 20% lower when administered during HD with LP dialyzers and approximately 35% lower with HP dialyzers. CONCLUSIONS: Intradialytic daptomycin administration during the last 30 min of HD is feasible, provided that larger dosages are used to compensate for intradialytic drug loss. On the basis of our findings, intradialytic doses of approximately 7 mg/kg (LP) or approximately 9 mg/kg (HP) theoretically should be bioequivalent to 6 mg/kg infused after HD. The calculated dosages are mathematically driven and must be validated in prospective clinical trials. PMID- 19541814 TI - Half of kidney transplant candidates who are older than 60 years now placed on the waiting list will die before receiving a deceased-donor transplant. AB - BACKGROUND AND OBJECTIVES: Waiting times to deceased-donor transplantation (DDTx) have significantly increased in the past decade. This trend particularly affects older candidates given a high mortality rate on dialysis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We conducted a retrospective analysis from the national Scientific Registry of Transplant Recipients database that included 54,669 candidates who were older than 60 yr and listed in the United States for a solitary kidney transplant from 1995 through 2007. Using survival models, we estimated time to DDTx and mortality after candidate listing with and without patients initially listed as temporarily inactive (status 7). RESULTS: Almost half (46%) of candidates who were older than 60 yr and listed in 2006 through 2007 are projected to die before receiving a DDTx. This proportion varied by individual characteristics: Diabetes (61%), age > or =70 yr (52%), black (62%), blood types O (60%) and B (71%), highly sensitized (68%), and on dialysis at listing (53%). Marked variation also existed by United Network for Organ Sharing region (6 to 81%). The overall projected proportion was reduced to 35% excluding patients who initially were listed as status 7. CONCLUSIONS: These data highlight the prominent and growing challenge facing the field of kidney transplantation. Older candidates are now at significant risk for not surviving the interval in which a deceased-donor transplant would become available. Importantly, this risk is variable within this population, and specific information should be disseminated to patients and caregivers to facilitate informed decision-making and potential incentives to seek living donors. PMID- 19541813 TI - HFE mutations modulate the effect of iron on serum hepcidin-25 in chronic hemodialysis patients. AB - BACKGROUND AND OBJECTIVES: Increased serum hepcidin has been reported in patients receiving chronic hemodialysis, and hypothesized to contribute to the alterations of iron metabolism of end-stage renal disease. However, no quantitative assessment is available to date; the clinical determinants are still under definition; and the role of genetic factors, namely HFE mutations, has not yet been evaluated. The aim of this study was to quantitatively assess serum hepcidin 25 in hemodialysis patients versus controls, and analyze the relationship between hepcidin, iron indices, HFE genotype, and erythropoietic parameters. DESIGN, SETTING, PARTICIPANTS & MEASUREMENTS: Sixty-five hemodialysis patients and 57 healthy controls were considered. Hepcidin-25 was evaluated by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, HFE genotype by restriction analysis. RESULTS: Serum hepcidin-25 was higher in hemodialysis patients compared with controls. In patients, hepcidin-25 correlated positively with ferritin and C reactive protein, and negatively with serum iron after adjustment for confounders. Hepcidin/ferritin ratio was lower in patients with (n = 25) than in those without (n = 40) HFE mutations. At multivariate analysis, hepcidin-25 was independently associated with ferritin and HFE status. In a subgroup of 22 "stable" patients, i.e., with Hb levels on target, normal CRP levels, and absence of complications for at least 1 yr, hepcidin-25 was negatively correlated with Hb levels independently of confounders. CONCLUSIONS: Serum hepcidin-25 is increased in hemodialysis patients, regulated by iron stores and inflammation, and relatively reduced in subjects carrying frequent HFE mutations. Hepcidin-25 may contribute to the pathogenesis of anemia by decreasing iron availability. PMID- 19541815 TI - Encapsulating peritoneal sclerosis in the new millennium: a national cohort study. AB - BACKGROUND AND OBJECTIVES: The study aim was to establish the incidence and characterize all encapsulating peritoneal sclerosis (EPS) cases in patients treated by peritoneal dialysis (PD). DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: The patient cohort, which started PD from January 1, 2000, to December 31, 2007, was identified from the Scottish Renal Registry (n = 1238). Possible EPS cases were identified by the ten adult Scottish renal units. Patient records were examined to ensure cases met diagnostic criteria. RESULTS: Forty-six cases were identified; 19 had their first PD exposure after January 1, 2000. The rate was 1.5%, an incidence of 4.9 per 1000 person-years. The incidence increased with PD duration, with rates of 0, 0.6, 2.0, 3.5, 8.1, 8.8 and 5% at <1, 1 to 2, >2 to 3, >3 to 4, >4 to 5, >5 to 6 and >6 yr PD exposure, respectively. The median PD duration of EPS cases was 5.1 yr (interquartile range [IQR] 3.4 to 6.1 yr). At diagnosis, 12 (26%) were on PD and 33 (72%) were diagnosed <2 yr after PD stopped. The cases had a median of 3.3 episodes of peritonitis (range 0 to 20, IQR 1 to 4.5). Thirty (65%) had used 3.86% dextrose dialysate and 45 (98%) had used Extraneal. The mortality was 42% at 1 yr postdiagnosis with a median survival of 149 d (IQR 61 to 408 d). CONCLUSIONS: The incidence reported in this study may be used to inform patients of the minimum risk of developing EPS on PD. PMID- 19541816 TI - Inflammation in renal transplantation. AB - BACKGROUND AND OBJECTIVES: Renal transplant recipients experience premature cardiovascular disease and death. The association of inflammation, all-cause mortality, and cardiovascular events in renal transplant recipients has not been examined in a large prospective controlled trial. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: ALERT was a randomized, double-blind, placebo-controlled study of the effect of fluvastatin on cardiovascular and renal outcomes in 2102 renal transplant recipients. Patients initially randomized to fluvastatin or placebo in the 5- to 6-yr trial were offered open-label fluvastatin in a 2-yr extension to the original study. The association between inflammation markers, high sensitivity C-reactive protein (hsCRP), and IL-6 on cardiovascular events and all cause mortality was investigated. RESULTS: The baseline IL-6 value was 2.9 +/- 1.9 pg/ml (n = 1751) and that of hsCRP was 3.8 +/- 6.7 mg/L (n = 1910). After adjustment for baseline values for established risk factors, the hazard ratios for a major cardiac event and all-cause mortality for IL-6 were 1.08 [95% confidence interval (CI), 1.01 to 1.15, P = 0.018] and 1.11 (95% CI, 1.05 to 1.18, P < 0.001), respectively. The adjusted hazard ratio for hsCRP for a cardiovascular event was 1.10 (95% CI, 1.01 to 1.20, P = 0.027) and for all-cause mortality was 1.15 (95% CI, 1.06 to 1.1.25, P = 0.049). CONCLUSIONS: The inflammation markers IL-6 and hsCRP are independently associated with major cardiovascular events and all-cause mortality in renal transplant recipients. PMID- 19541817 TI - Sensitivity of billing claims for cardiovascular disease events among kidney transplant recipients. AB - BACKGROUND AND OBJECTIVES: Billing claims are increasingly examined beyond administrative functions as outcomes measures in observational research. Few studies have described the performance of billing claims as surrogate measures of clinical events among kidney transplant recipients. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We investigated the sensitivity of Medicare billing claims for clinically verified cardiovascular diagnoses (five categories) and procedures (four categories) in a novel database linking Medicare claims to electronic medical records of one transplant program. Cardiovascular events identified in medical records for 571 Medicare-insured transplant recipients in 1991 through 2002 served as reference measures. RESULTS: Within a claims ascertainment period spanning +/-30 d of clinically recorded dates, aggregate sensitivity of single claims was higher for case definitions incorporating Medicare Parts A and B for diagnoses and procedures (90.9%) compared with either Part A (82.3%) or Part B (84.6%) alone. Perfect capture of the four procedures was possible within +/-30 d or with short claims window expansion, but sensitivity for the diagnoses trended lower with all study algorithms (91.2% with window up to +/-90 d). Requirement for additional confirmatory diagnosis claims did not appreciably reduce sensitivity. Sensitivity patterns were similar in the early compared with late periods of the study. CONCLUSIONS: Combined use of Medicare Parts A and B billing claims composes a sensitive measure of cardiovascular events after kidney transplant. Further research is needed to define algorithms that maximize specificity as well as sensitivity of claims from Medicare and other insurers as research measures in this population. PMID- 19541819 TI - Leukocyte proteases cleave von Willebrand factor at or near the ADAMTS13 cleavage site. AB - The function of von Willebrand factor (VWF) is regulated by proteolysis, which limits its multimeric size and ability to tether platelets. The importance of ADAMTS13 metalloprotease in VWF regulation is demonstrated by the association between severe deficiency of ADAMTS13 and thrombotic thrombocytopenic purpura (TTP). However, ADAMTS13 activity levels do not always correlate with the clinical course of TTP, suggesting that other proteases could be important in regulating VWF. We identified 4 leukocyte proteases that cleave the synthetic VWF substrate FRETS-VWF73 and multimeric VWF. Elastase and proteinase 3 (PR3) cleave multimeric VWF and FRETS-VWF73 at the V(1607)-T(1608) peptide bond; cathepsin G and matrix metalloprotease 9 cleave VWF substrates at the Y(1605)-M(1606) and M(1606)-V(1607) bonds, respectively. Isolated intact human neutrophils cleave FRETS-VWF73 at the V(1607)-T(1608) peptide bond, suggesting that elastase or PR3 expressed on leukocyte surfaces might cleave VWF. In the presence of normal or ADAMTS13-deficient plasma, cleavage of FRETS-VWF73 by resting neutrophils is abolished. However, activated neutrophils retain proteolytic activity toward FRETS-VWF73 in the presence of plasma. Although the in vivo relevance remains to be established, these studies suggest the existence of a "hot spot" of VWF proteolysis in the VWF A2 domain, and support the possibility that activated leukocytes may participate in the proteolytic regulation of VWF. PMID- 19541818 TI - Chronic Renal Insufficiency Cohort (CRIC) Study: baseline characteristics and associations with kidney function. AB - BACKGROUND AND OBJECTIVES: The Chronic Renal Insufficiency Cohort (CRIC) Study was established to examine risk factors for the progression of chronic kidney disease (CKD) and cardiovascular disease (CVD) in patients with CKD. We examined baseline demographic and clinical characteristics. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Seven clinical centers recruited adults who were aged 21 to 74 yr and had CKD using age-based estimated GFR (eGFR) inclusion criteria. At baseline, blood and urine specimens were collected and information regarding health behaviors, diet, quality of life, and functional status was obtained. GFR was measured using radiolabeled iothalamate in one third of participants. RESULTS: A total of 3612 participants were enrolled with mean age +/- SD of 58.2 +/- 11.0 yr; 46% were women, and 47% had diabetes. Overall, 45% were non-Hispanic white, 46% were non-Hispanic black, and 5% were Hispanic. Eighty-six percent reported hypertension, 22% coronary disease, and 10% heart failure. Mean body mass index was 32.1 +/- 7.9 kg/m(2), and 47% had a BP >130/80 mmHg. Mean eGFR was 43.4 +/- 13.5 ml/min per 1.73 m(2), and median (interquartile range) protein excretion was 0.17 g/24 h (0.07 to 0.81 g/24 h). Lower eGFR was associated with older age, lower socioeconomic and educational level, cigarette smoking, self-reported CVD, peripheral arterial disease, and elevated BP. CONCLUSIONS: Lower level of eGFR was associated with a greater burden of CVD as well as lower socioeconomic and educational status. Long-term follow-up of participants will provide critical insights into the epidemiology of CKD and its relationship to adverse outcomes. PMID- 19541820 TI - In essential thrombocythemia, multiple JAK2-V617F clones are present in most mutant-positive patients: a new disease paradigm. AB - In essential thrombocythemia (ET), the JAK2-V617F mutation is usually restricted to a subpopulation of neutrophils and platelets, and production of JAK2 wild-type (WT) platelets is not suppressed. Nonmutated precursor cells may, therefore, be susceptible to the acquisition of further JAK2 mutations. We used a common single nucleotide polymorphism (SNP) in the JAK2 coding sequence to genotype V617F alleles obtained either by allele-specific restriction enzyme digestion (RED) or by cloning. Both SNP alleles were detected in JAK2 mutant-positive alleles from neutrophils of 10 of 11 ET patients studied using RED compared with 0 of 5 with polycythemia vera. These results were confirmed in cloned products from 5 ET patients and indicate the occurrence of at least 2 separate JAK2 mutation events in the majority of ET patients investigated. In a further ET patient, JAK2 mutant positive erythroid colonies with either X-allele inactivated were detected, demonstrating they could not have arisen from a common clonal precursor. These results indicate that at least 2 independent JAK2-V617F events occur commonly in ET patients, and they may arise on a polyclonal background. The presence of a JAK2 mutation in ET patients should not, therefore, be equated with a malignant disease. PMID- 19541821 TI - Restoration of NET formation by gene therapy in CGD controls aspergillosis. AB - Chronic granulomatous disease (CGD) patients have impaired nicotinamide adenine dinucleotide phosphate (NADPH) oxidase function, resulting in poor antimicrobial activity of neutrophils, including the inability to generate neutrophil extracellular traps (NETs). Invasive aspergillosis is the leading cause of death in patients with CGD; it is unclear how neutrophils control Aspergillus species in healthy persons. The aim of this study was to determine whether gene therapy restores NET formation in CGD by complementation of NADPH oxidase function, and whether NETs have antimicrobial activity against Aspergillus nidulans. Here we show that reconstitution of NET formation by gene therapy in a patient with CGD restores neutrophil elimination of A nidulans conidia and hyphae and is associated with rapid cure of preexisting therapy refractory invasive pulmonary aspergillosis, underlining the role of functional NADPH oxidase in NET formation and antifungal activity. PMID- 19541822 TI - Forodesine has high antitumor activity in chronic lymphocytic leukemia and activates p53-independent mitochondrial apoptosis by induction of p73 and BIM. AB - Chronic lymphocytic leukemia (CLL) is an incurable disease derived from the monoclonal expansion of CD5(+) B lymphocytes. High expression levels of ZAP-70 or CD38 and deletions of 17p13 (TP53) and 11q22-q23 (ATM) are associated with poorer overall survival and shorter time to disease progression. DNA damage and p53 play a pivotal role in apoptosis induction in response to conventional chemotherapy, because deletions of ATM or p53 identify CLL patients with resistance to treatment. Forodesine is a transition-state inhibitor of the purine nucleoside phosphorylase with antileukemic activity. We show that forodesine is highly cytotoxic as single agent or in combination with bendamustine and rituximab in primary leukemic cells from CLL patients regardless of CD38/ZAP-70 expression and p53 or ATM deletion. Forodesine activates the mitochondrial apoptotic pathway by decreasing the levels of antiapoptotic MCL-1 protein and induction of proapoptotic BIM protein. Forodesine induces transcriptional up-regulation of p73, a p53-related protein able to overcome the resistance to apoptosis of CLL cells lacking functional p53. Remarkably, no differences in these apoptotic markers were observed based on p53 or ATM status. In conclusion, forodesine induces apoptosis of CLL cells bypassing the DNA-damage/ATM/p53 pathway and might represent a novel chemotherapeutic approach that deserves clinical investigation. PMID- 19541823 TI - KW-2449, a novel multikinase inhibitor, suppresses the growth of leukemia cells with FLT3 mutations or T315I-mutated BCR/ABL translocation. AB - KW-2449, a multikinase inhibitor of FLT3, ABL, ABL-T315I, and Aurora kinase, is under investigation to treat leukemia patients. In this study, we examined its possible modes of action for antileukemic effects on FLT3-activated, FLT3 wild type, or imatinib-resistant leukemia cells. KW-2449 showed the potent growth inhibitory effects on leukemia cells with FLT3 mutations by inhibition of the FLT3 kinase, resulting in the down-regulation of phosphorylated-FLT3/STAT5, G(1) arrest, and apoptosis. Oral administration of KW-2449 showed dose-dependent and significant tumor growth inhibition in FLT3-mutated xenograft model with minimum bone marrow suppression. In FLT3 wild-type human leukemia, it induced the reduction of phosphorylated histone H3, G(2)/M arrest, and apoptosis. In imatinib resistant leukemia, KW-2449 contributed to release of the resistance by the simultaneous down-regulation of BCR/ABL and Aurora kinases. Furthermore, the antiproliferative activity of KW-2449 was confirmed in primary samples from AML and imatinib-resistant patients. The inhibitory activity of KW-2449 is not affected by the presence of human plasma protein, such as alpha1-acid glycoprotein. These results indicate KW-2449 has potent growth inhibitory activity against various types of leukemia by several mechanisms of action. Our studies indicate KW-2449 has significant activity and warrants clinical study in leukemia patients with FLT3 mutations as well as imatinib-resistant mutations. PMID- 19541824 TI - In vitro inhibition of multiple cytochrome P450 isoforms by xanthone derivatives from mangosteen extract. AB - Mangosteen is a xanthone-containing fruit found in Southeast Asia for which health claims include maintaining healthy immune and gastrointestinal systems to slowing the progression of tumor growth and neurodegenerative diseases. Previous studies have identified multiple xanthones in the pericarp of the mangosteen fruit. The aim of the current study was to assess the drug inhibition potential of mangosteen in vitro as well as the cytochrome P450 (P450) enzymes responsible for the metabolism of its individual components. The various xanthone derivatives were found to be both substrates and inhibitors for multiple P450 isoforms. Aqueous extracts of the mangosteen pericarp were analyzed for xanthone content as well as inhibition potency. Finally, in vivo plasma concentrations of alpha mangostin, the most abundant xanthone derivative found in mangosteen, were predicted using Simcyp and found to be well above their respective in vitro K(i) values for CYP2C8 and CYP2C9. PMID- 19541825 TI - Aromatic substitution reaction of 2-chloropyridines catalyzed by microsomal glutathione S-transferase 1. AB - We investigated the substitution reaction of a series of 2-chloropyridine derivatives catalyzed by rat liver microsomal glutathione S-transferase 1. Various 2-chloropyridine derivatives were metabolized to the corresponding substituted glutathione conjugates via displacement of chlorine atom with glutathione. The reaction was affected by the electron-withdrawing strength and position of the substituents. Molecular orbital calculations on the change in Gibbs free energy between the initial and transition states verified the presence of a Meisenheimer complex and its influence on the reaction rate. PMID- 19541826 TI - Contrasting influence of NADPH and a NADPH-regenerating system on the metabolism of carbonyl-containing compounds in hepatic microsomes. AB - Carbonyl containing xenobiotics may be susceptible to NADPH-dependent cytochrome P450 (P450) and carbonyl-reduction reactions. In vitro hepatic microsome assays are routinely supplied NADPH either by direct addition of NADPH or via an NADPH regenerating system (NRS). In contrast to oxidative P450 transformations, which occur on the periphery of a microsome vesicle, intraluminal carbonyl reduction depends on transport of cofactors across the endoplasmic reticulum (ER) membrane into the lumen. Glucose 6-phosphate, a natural cofactor and component of the NRS matrix, is readily transported across the ER membrane and facilitates intraluminal NADPH production, whereas direct addition of NADPH has limited access to the lumen. In this study, we compared the effects of direct addition of NADPH and use of an NRS on the P450-mediated transformation of propiconazole and 11 beta-hydroxysteroid dehydrogenase type 1 (HSD1) carbonyl reduction of cortisone and the xenobiotic triadimefon in hepatic microsomes. Our results demonstrate that the use of NADPH rather than NRS can underestimate the kinetic rates of intraluminal carbonyl reduction, whereas P450-mediated transformations were unaffected. Therefore, in vitro depletion rates measured for a carbonyl containing xenobiotic susceptible to both intraluminal carbonyl reduction and P450 processes may not be properly assessed with direct addition of NADPH. In addition, we used in silico predictions as follows: 1) to show that 11 beta-HSD1 carbonyl reduction was energetically more favorable than oxidative P450 transformation; and 2) to calculate chemical binding score and the distance between the carbonyl group and the hydride to be transferred by NADPH to identify other 11 beta-HSD1 substrates for which reaction kinetics may be underestimated by direct addition of NADPH. PMID- 19541827 TI - Metabolism of ticlopidine in rats: identification of the main biliary metabolite as a glutathione conjugate of ticlopidine S-oxide. AB - We have identified several novel metabolites of ticlopidine, a well known antiplatelet agent and have revealed its metabolic route in rats. The main biliary metabolite of ticlopidine was characterized as a glutathione (GSH) conjugate of ticlopidine S-oxide, in which conjugation had occurred at carbon 7a in the thienopyridine moiety. Quantitative analysis revealed that 29% of the dose was subjected to the formation of reactive intermediates followed by conjugation with GSH after oral administration of ticlopidine (22 mg/kg) to rats. In vitro incubation of ticlopidine with rat liver 9000 g supernatant fraction (S9) fractions led to the formation of multiple metabolites, including 2-oxo ticlopidine, the precursor for the pharmacologically active ticlopidine metabolite, [1-(2-chlorobenzyl)-4-mercaptopiperidin-(3Z)-ylidene] acetic acid. A novel thiophene ring-opened metabolite with a thioketone group and a carboxylic acid moiety has also been detected after incubation of 2-oxo-ticlopidine with rat liver microsomes or upon incubation of ticlopidine with rat liver S9 fractions. PMID- 19541828 TI - Genetic polymorphisms in the TATA box and upstream phenobarbital-responsive enhancer module of the UGT1A1 promoter have combined effects on UDP glucuronosyltransferase 1A1 transcription mediated by constitutive androstane receptor, pregnane X receptor, or glucocorticoid receptor in human liver. AB - Transcription of UDP-glucuronosyltransferase (UGT) 1A1 is regulated by the transcription factors, constitutive androstane receptor (CAR), pregnane X receptor (PXR), glucocorticoid receptor (GR), hepatocyte nuclear factor (HNF) 1 alpha, and HNF4 alpha. The purpose of this study was to determine whether the genetic polymorphisms in the RNA polymerase II core promoter and the upstream phenobarbital-responsive element module (PBREM) of the UGT1A1 promoter have combined effects on UGT1A1 transcription mediated by the transcription factors. A polymorphism of A(TA)(5-8)TAA in the UGT1A1 TATA box and a single nucleotide polymorphism of -3279T>G in PBREM were genotyped in 98 human liver samples. Relative mRNA levels of CAR, PXR, GR, HNF1 alpha, HNF4 alpha, and UGT1A1 were quantified by a multiplex branched DNA technique. Correlations of mRNA levels between UGT1A1 and the transcription factors were established in liver samples with different combined genetic polymorphisms. Correlation of mRNA levels between UGT1A1 and CAR, PXR, or GR, but not HNF1 alpha or HNF4 alpha, was abolished in the samples with the combined genotype of TA7/7 plus -3279G/G, which was also associated with significantly lower UGT1A1 mRNA levels compared with other combined genotypes. Correlations of mRNA levels between UGT1A1 and CAR or PXR were reduced but not abolished in the samples with the combined genotype of TA6/7 plus -3279 G/G, which showed significantly lower UGT1A1 mRNA levels compared with the combined genotype of TA6/7 plus -3279T/G and other genotypes containing TA6/6. In conclusion, the combined genotypes containing A(TA)(7)TAA and -3279G decrease UGT1A transcription mediated by CAR, PXR, or GR but not by HNF1 alpha or HNF4 alpha. PMID- 19541829 TI - Substrate-dependent functional alterations of seven CYP2C9 variants found in Japanese subjects. AB - CYP2C9 is a polymorphic enzyme that metabolizes a number of clinically important drugs. In this study, catalytic activities of seven alleles found in Japanese individuals, CYP2C9*3 (I359L), *13 (L90P), *26 (T130R), *28 (Q214L), *30 (A477T), *33 (R132Q), and *34 (R335Q), were assessed using three substrates (diclofenac, losartan, and glimepiride). When expressed in a baculovirus-insect cell system, the holo and total (apo and holo) CYP2C9 protein expression levels were similar among the wild type (CYP2C9.1) and six variants except for CYP2C9.13. A large part of CYP2C9.13 was present in the apo form P420. Compared with CYP2C9.1, all variants except for CYP2C9.34 exhibited substrate-dependent changes in K(m), V(max), and intrinsic clearance (V(max)/K(m)). For diclofenac 4'-hydroxylation, the intrinsic clearance was decreased markedly (by >80%) in CYP2C9.13, CYP2C9.30, and CYP2C9.33 and variably (63-76%) in CYP2C9.3, CYP2C9.26, and CYP2C9.28 due to increased K(m) and/or decreased V(max) values. For losartan oxidation, CYP2C9.13 and CYP2C9.28 showed 2.5- and 1.8-fold higher K(m) values, respectively, and all variants except for CYP2C9.34 showed >77% lower V(max) and intrinsic clearance values. For glimepiride hydroxylation, the K(m) of CYP2C9.13 was increased 7 fold, and the V(max) values of all variants significantly decreased, resulting in reductions in the intrinsic clearance by >80% in CYP2C9.3, CYP2C9.13, CYP2C9.26, and CYP2C9.33 and by 56 to 75% in CYP2C9.28 and CYP2C9.30. These findings suggest the necessity for careful administration of losartan and glimepiride to patients bearing these six alleles. PMID- 19541830 TI - Flt3L, DCs, and NTregs: team contra GVHD? PMID- 19541831 TI - NKT cells in p53 deficiency. PMID- 19541832 TI - MicroRNA: mIR-ly regulators of DNMT? PMID- 19541833 TI - Babies born without safety NET. PMID- 19541834 TI - Escaping the niche. PMID- 19541835 TI - Dissecting VHL-associated pathologies. PMID- 19541836 TI - Is a little GVHD a good thing? PMID- 19541837 TI - The proposed diagnostic criteria change for chronic lymphocytic leukemia: unintended consequences? PMID- 19541838 TI - Lymphocytes, B lymphocytes, and clonal CLL cells: observations on the impact of the new diagnostic criteria in the 2008 Guidelines for Chronic Lymphocytic Leukemia (CLL). PMID- 19541840 TI - p73, miR106b, miR34a, and Itch in chronic lymphocytic leukemia. PMID- 19541842 TI - MicroRNA expression in human airway smooth muscle cells: role of miR-25 in regulation of airway smooth muscle phenotype. AB - Defining mechanisms by which differentiated, contractile smooth muscle cells become proliferative and secretory in response to mechanical and environmental stress is crucial for determining the contribution of airway smooth muscle (ASM) to inflammatory responses that result in airway disease. Regulation by microRNAs (miRNAs) has emerged as an important post-transcriptional mechanism regulating gene expression that may modulate ASM phenotype, but little is known about the expression and functions of miRNA in smooth muscle. In the present study we used microarrays to determine whether miRNAs in human ASM cells are altered by a proinflammatory stimulus. In ASM cells exposed to IL-1beta, TNF-alpha, and IFN gamma, we found 11 miRNAs to be significantly down-regulated. We verified decreased expression of miR-25, miR-140*, mir-188, and miR-320 by quantitative PCR. Analysis of miR-25 expression indicates that it has a broad role in regulating ASM phenotype by modulating expression of inflammatory mediators such as RANTES, eotaxin, and TNF-alpha; genes involved in extracellular matrix turnover; and contractile proteins, most notably myosin heavy chain. miRNA binding algorithms predict that miR-25 targets Kruppel-like factor 4 (KLF4), a potent inhibitor of smooth muscle-specific gene expression and mediator of inflammation. Our study demonstrates that inhibition of miR-25 in cytokine stimulated ASM cells up-regulates KLF4 expression via a post-transcriptional mechanism. This provides novel evidence that miR-25 targets KLF4 in ASM cells and proposes that miR-25 may be an important mediator of ASM phenotype. PMID- 19541843 TI - Role of the serotonergic system in reduced pulmonary function after exposure to methamphetamine. AB - Although use of methamphetamine (MA) by smoking is the fastest growing method of administration, very limited data are available describing the effects of smoked MA. Using a murine inhalation exposure system, we explored the pulmonary effects of low-dose acute inhalation exposure to MA vapor (smoke). Inhalation of MA vapor resulted in transiently reduced pulmonary function, as measured by transpulmonary resistance, dynamic compliance, and whole-body plethysmography compared with unexposed control animals. These changes were associated with an approximately 34% reduction in serotonin (5-hydroxytryptamine [5-HT]) metabolism/inactivation to 5-hydroxyindolacetic acid, and a nearly 40% reduction in monoamine oxidase (MAO)-A activity in the lung. Pretreatment of mice with a selective 5-HT reuptake inhibitor completely ablated the MA-induced changes in pulmonary function, confirming a key role for the 5-HT transporter (serotonin transporter [SERT]) and the serotonergic system in this effect. Immunofluorescent staining of mouse lung tissue confirmed high expression of SERT in airway epithelial cells. Using mouse airway epithelial cell line, LA-4, and purified human MAO-A, it was demonstrated that MA impedes 5-HT metabolism through direct inhibition of MAO-A activity in vitro. Together, these data demonstrate that low-dose exposure to MA results in reduced pulmonary function mediated via SERT and subsequent perturbation of 5-HT metabolism in the lung. This supports a role for the serotonergic system in MA mediated pulmonary effects. PMID- 19541845 TI - Receptors for advanced glycation end-products targeting protect against hyperoxia induced lung injury in mice. AB - Patients with acute lung injury almost always require supplemental oxygen during treatment; however, elevated oxygen itself is toxic. Receptors for advanced glycation end-products (RAGE) are multi-ligand cell surface receptors predominantly localized to alveolar type I cells that influence development and cigarette smoke-induced inflammation, but studies that address the role of RAGE in acute lung injury are insufficient. In the present investigation, we test the hypothesis that RAGE signaling functions in hyperoxia-induced inflammation. RAGE null mice exposed to hyperoxia survived 3 days longer than age-matched wild-type mice. After 4 days in hyperoxia, RAGE-null mice had less total cell infiltration into the airway, decreased total protein leak, diminished alveolar damage in hematoxylin and eosin-stained lung sections, and a lower lung wet-to-dry weight ratio. An inflammatory cytokine antibody array revealed decreased secretion of several proinflammatory molecules in lavage fluid obtained from RAGE knockout mice when compared with wild-type control animals. Real-time RT-PCR and immunoblotting revealed that hyperoxia induced RAGE expression in primary alveolar epithelial cells, and immunohistochemistry identified increased RAGE expression in the lungs of mice after exposure to hyperoxia. These data reveal that RAGE targeting leads to a diminished hyperoxia-induced pulmonary inflammatory response. Further research into the role of RAGE signaling in the lung should identify novel targets likely to be important in the therapeutic alleviation of lung injury and associated persistent inflammation. PMID- 19541844 TI - Conditional overexpression of connective tissue growth factor disrupts postnatal lung development. AB - Connective tissue growth factor (CTGF) is a member of an emerging family of immediate-early gene products that coordinates complex biological processes during development, differentiation, and tissue repair. Overexpression of CTGF is associated with mechanical ventilation with high tidal volume and oxygen exposure in newborn lungs. However, the role of CTGF in postnatal lung development and remodeling is not well understood. In the present study, a double-transgenic mouse model was generated with doxycycline-inducible overexpression of CTGF in respiratory epithelial cells. Overexpression of CTGF from Postnatal Days 1-14 resulted in thicker alveolar septa and decreased secondary septal formation. This is correlated with increased myofibroblast differentiation and disorganized elastic fiber deposition in alveolar septa. Overexpression of CTGF also decreased alveolar capillary network formation. There were increased alpha-smooth muscle actin expression and collagen deposition, and dramatic thickening in the peribronchial/peribronchiolar and perivascular regions in the double-transgenic lungs. Furthermore, overexpression of CTGF increased integrin-linked kinase expression, activated its downstream signaling target, Akt, as well as increased mRNA expression of fibronectin. These data demonstrate that overexpression of CTGF disrupts alveologenesis and capillary formation, and induces fibrosis during the critical period of alveolar development. These histologic changes are similar to those observed in lungs of infants with bronchopulmonary dysplasia. PMID- 19541846 TI - Latissimus dorsi and teres major tears in professional baseball pitchers: a case series. AB - BACKGROUND: Latissimus dorsi and teres major tendon tears are uncommon injuries. Only a few case reports exist, mainly in high-level athletes. PURPOSE: To describe a series of latissimus dorsi and teres major tendon tears in professional baseball pitchers. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Injury data from 3 Major League Baseball clubs were collected over a total of 10 seasons. Any baseball player who sustained an injury to either the latissimus dorsi or teres major identified on magnetic resonance imaging (MRI) was included. All injured players were treated nonoperatively with a goal of returning to full velocity throwing at 3 months from the time of injury. RESULTS: Ten players sustained injuries to the latissimus dorsi and/or the teres major during pitching. The MRI findings documented 5 isolated latissimus dorsi tears, 4 isolated teres major tears, and 1 combined injury. All athletes returned to pitching, and all but 1 player returned to baseball at the same level of competition in the same season. Nine of 10 players returned at 3 months from the time of their injury. One recurrence was seen 6 months after returning to throwing; however, this healed with further nonoperative treatment, and the player returned to competition at the same level 6 weeks later. One player had continued shoulder symptoms and retired at the end of the season. CONCLUSION: Although uncommon, tears of the latissimus dorsi and teres major occur in professional baseball players. Acute injuries are demonstrated on standard shoulder MRI, although larger field-of-view images are required to accurately assess the injury. Most heal successfully with nonoperative treatment, and most players are able to return to the same level of competition in 3 months. PMID- 19541847 TI - Delayed foreign body reaction due to bioabsorbable pins used for femoral fixation in anterior cruciate ligament reconstruction: a case report. PMID- 19541848 TI - Ablation of adhesion molecule L1 in mice favours Schwann cell proliferation and functional recovery after peripheral nerve injury. AB - The adhesion molecule L1 is one of the few adhesion molecules known to be beneficial for repair processes in the adult central nervous system of vertebrates by promoting axonal growth and neuronal survival. In the peripheral nervous system, L1 is up-regulated by myelination-competent Schwann cells and regenerating axons after nerve damage but its functional role has remained unknown. Here we tested the hypothesis that L1 is, as in the central nervous system, beneficial for nerve regeneration in the peripheral nervous system by performing combined functional and histological analyses of adult L1-deficient mice (L1y/-) and wild-type (L1y/+) littermates. Contrary to our hypothesis, quantitative video-based motion analysis revealed better locomotor recovery in L1y/- than in L1y/+ mice at 4-12 weeks after transection and surgical repair of the femoral nerve. Motoneuron regeneration in L1y/- mice was also enhanced as indicated by attenuated post-traumatic loss of motoneurons, enhanced precision of motor reinnervation, larger cell bodies of regenerated motoneurons and diminished loss of inhibitory synaptic input to motoneurons. In search of mechanisms underlying the observed effects, we analysed peripheral nerves at short time periods (3-14 days) after transection and found that Schwann cell proliferation is strongly augmented in L1y/- versus L1y/+ mice. L1-deficient Schwann cells showed increased proliferation than wild-type Schwann cells, both in vivo and in vitro. These findings suggest a novel role for L1 in nerve regeneration. We propose that L1 negatively regulates Schwann cell proliferation after nerve damage, which in turn restricts functional recovery by limiting the trophic support for regenerating motoneurons. PMID- 19541849 TI - Identification of small gains and losses in single cells after whole genome amplification on tiling oligo arrays. AB - Clinical DNA is often available in limited quantities requiring whole-genome amplification for subsequent genome-wide assessment of copy-number variation (CNV) by array-CGH. In pre-implantation diagnosis and analysis of micrometastases, even merely single cells are available for analysis. However, procedures allowing high-resolution analyses of CNVs from single cells well below resolution limits of conventional cytogenetics are lacking. Here, we applied amplification products of single cells and of cell pools (5 or 10 cells) from patients with developmental delay, cancer cell lines and polar bodies to various oligo tiling array platforms with a median probe spacing as high as 65 bp. Our high-resolution analyses reveal that the low amounts of template DNA do not result in a completely unbiased whole genome amplification but that stochastic amplification artifacts, which become more obvious on array platforms with tiling path resolution, cause significant noise. We implemented a new evaluation algorithm specifically for the identification of small gains and losses in such very noisy ratio profiles. Our data suggest that when assessed with sufficiently sensitive methods high-resolution oligo-arrays allow a reliable identification of CNVs as small as 500 kb in cell pools (5 or 10 cells), and of 2.6-3.0 Mb in single cells. PMID- 19541850 TI - Activation of interferon regulatory factor-3 via toll-like receptor 3 and immunomodulatory functions detected in A549 lung epithelial cells exposed to misplaced U1-snRNA. AB - U1-snRNA is an integral part of the U1 ribonucleoprotein pivotal for pre-mRNA splicing. Toll-like receptor (TLR) signaling has recently been associated with immunoregulatory capacities of U1-snRNA. Using lung A549 epithelial/carcinoma cells, we report for the first time on interferon regulatory factor (IRF)-3 activation initiated by endosomally delivered U1-snRNA. This was associated with expression of the IRF3-inducible genes interferon-beta (IFN-beta), CXCL10/IP-10 and indoleamine 2,3-dioxygenase. Mutational analysis of the U1-snRNA-activated IFN-beta promoter confirmed the crucial role of the PRDIII element, previously proven pivotal for promoter activation by IRF3. Notably, expression of these parameters was suppressed by bafilomycin A(1), an inhibitor of endosomal acidification, implicating endosomal TLR activation. Since resiquimod, an agonist of TLR7/8, failed to stimulate A549 cells, data suggest TLR3 to be of prime relevance for cellular activation. To assess the overall regulatory potential of U1-snRNA-activated epithelial cells on cytokine production, co-cultivation with peripheral blood mononuclear cells (PBMC) was performed. Interestingly, A549 cells activated by U1-snRNA reinforced phytohemagglutinin-induced interleukin-10 release by PBMC but suppressed that of tumor necrosis factor-alpha, indicating an anti-inflammatory potential of U1-snRNA. Since U1-snRNA is enriched in apoptotic bodies and epithelial cells are capable of performing efferocytosis, the present data in particular connect to immunobiological aspects of apoptosis at host/environment interfaces. PMID- 19541851 TI - Establishment of histone modifications after chromatin assembly. AB - Every cell has to duplicate its entire genome during S-phase of the cell cycle. After replication, the newly synthesized DNA is rapidly assembled into chromatin. The newly assembled chromatin 'matures' and adopts a variety of different conformations. This differential packaging of DNA plays an important role for the maintenance of gene expression patterns and has to be reliably copied in each cell division. Posttranslational histone modifications are prime candidates for the regulation of the chromatin structure. In order to understand the maintenance of chromatin structures, it is crucial to understand the replication of histone modification patterns. To study the kinetics of histone modifications in vivo, we have pulse-labeled synchronized cells with an isotopically labeled arginine ((15)N(4)) that is 4 Da heavier than the naturally occurring (14)N(4) isoform. As most of the histone synthesis is coupled with replication, the cells were arrested at the G1/S boundary, released into S-phase and simultaneously incubated in the medium containing heavy arginine, thus labeling all newly synthesized proteins. This method allows a comparison of modification patterns on parental versus newly deposited histones. Experiments using various pulse/chase times show that particular modifications have considerably different kinetics until they have acquired a modification pattern indistinguishable from the parental histones. PMID- 19541852 TI - Revisiting the association between cardiovascular risk factors and diabetes: data from a large population-based study. AB - PURPOSE: The purpose of this study was to examine the association between cardiovascular risk factors and the presence of diabetes in a large population level dataset. METHODS: A secondary analysis was conducted using data from the 2007 Behavioral Risk Factor Surveillance System, a population-based survey (n = 403,137) conducted in the United States. RESULTS: The majority of the respondents were middle-aged and overweight. Approximately half of the sample reported little or no physical activity. Estimates from a logistic regression model for a weighted sample of white, black, and Hispanic adults revealed that having hypertension or elevated cholesterol was a strong predictor of diabetes even when controlling for age, gender, race, education, income, body mass index, smoking status, and physical activity. CONCLUSIONS: The results confirmed the importance of diabetes educators counseling patients with hypertension or hypercholesterolemia about their increased risk for developing diabetes. PMID- 19541853 TI - Increased skin carcinogenesis in caspase-activated DNase knockout mice. AB - Caspase-activated DNase (CAD), also called DNA fragmentation factor (DFF), is the enzyme responsible for DNA fragmentation during apoptosis, a hallmark of programmed cell death. CAD/DFF has been shown to suppress radiation-induced carcinogenesis by preventing genomic instability in cells. In this study, we have investigated the role of CAD in chemical carcinogenesis using CAD-null mice and two-stage model of skin carcinogenesis. After topical treatment of mouse skin with dimethylbenz[a]anthracene (DMBA) as an initiator and 12-O tetradecanoylphorbol-13-acetate (TPA) as a promoting agent, there was a 4-fold increase in the number of papillomas per mouse and 50.8% increase in the incidence of papilloma formation in the CAD knockout mice compared with wild-type littermates. The papillomas in CAD-null mice grew faster and reached larger sizes. These data indicate that loss of CAD function enhances tumorigenesis induced by a chemical carcinogen in the DMBA/TPA two-stage model of skin carcinogenesis in mice. PMID- 19541854 TI - Interferon-beta treatment increases human papillomavirus early gene transcription and viral plasmid genome replication by activating interferon regulatory factor (IRF)-1. AB - Interferons (IFNs) have been used to treat mucosal lesions caused by human papillomavirus (HPV) infection, such as intraepithelial precursor lesions to cancer of the uterine cervix, genital warts or recurrent respiratory papillomatosis, to potentially reduce or eliminate replicating HPV plasmid genomes. Mucosal HPVs have evolved mechanisms that impede IFN-beta synthesis and downregulate genes induced by IFN. Here we show that these HPV types directly subvert a cellular transcriptional response to IFN-beta as a potential boost in infection. Treatment with low levels of human IFN-beta induced initial amplification of HPV-16 and HPV-11 plasmid genomes and increased HPV-16 or HPV-31 DNA copy numbers up to 6-fold in HPV-immortalized keratinocytes. IFN treatment also increased early gene transcription from the major early gene promoters in HPV-16, HPV-31 and HPV-11. Furthermore, mutagenesis of the viral genomes and ectopic interferon regulatory factor (IRF) expression in transfection experiments using IRF-1(-/-), IRF-2(-/-) and dual knockout cell lines determined that these responses are due to the activation of IRF-1 interaction with a conserved interferon response element demonstrated in several mucosal HPV early gene promoters. Our results provide a molecular explanation for the varying clinical outcomes of IFN therapy of papillomatoses and define an assay for the modulation of the HPV gene program by IFNs as well as other cytokines and signaling molecules in infection and therapy. PMID- 19541855 TI - Effect of folic acid supplementation on the progression of colorectal aberrant crypt foci. AB - Whether or not folic acid supplementation promotes the progression of colorectal preneoplastic lesions to cancer is an important public health issue, given mandatory fortification and widespread supplemental use of folic acid in North America. We investigated the effect of folic acid supplementation on the progression of aberrant crypt foci (ACF), the earliest precursor of colorectal cancer. Male Sprague-Dawley rats (n = 152) were placed on a control diet (2 mg folic acid/kg diet) at weaning and ACF were induced by azoxymethane. Six weeks post-ACF induction, rats were randomized to receive 0, 2, 5 or 8 mg folic acid/kg diet. At 34 weeks of age, rats were killed, and colorectal tumor parameters, plasma folate and homocysteine (a sensitive inverse indicator of tissue folate status) concentrations and rectal epithelial proliferation were determined. Although the number of ACF increased as dietary folic acid levels increased (P = 0.015), the incidence of colorectal tumors did not differ significantly among the four dietary groups. However, tumor multiplicity was positively correlated with dietary folic acid levels (r = 0.32; P = 0.002) and inversely with plasma homocysteine concentrations (r = -0.32; P = 0.005). Tumor burden was positively correlated with dietary folic acid levels (r = 0.35; P = 0.001) and plasma folate concentrations (r = 0.33; P = 0.008) and inversely with plasma homocysteine concentrations (r = -0.42; P < 0.001). Rectal epithelial proliferation was positively correlated with dietary folic acid levels (r = 0.39; P < 0.001) and plasma folate concentrations (r = 0.34; P < 0.001) and inversely with plasma homocysteine concentrations (r = -0.37; P < 0.001). Our data suggest that folic acid supplementation may promote the progression of ACF to colorectal tumors. PMID- 19541856 TI - Life-years gained among US adults from modern treatments and changes in the prevalence of 6 coronary heart disease risk factors between 1980 and 2000. AB - Has the recent US decline in coronary heart disease (CHD) mortality increased life expectancy? The authors estimated the number of life-years gained from CHD treatments and changes in the prevalence of cardiovascular disease risk factors for the US population between 1980 and 2000. The previously validated IMPACT model was used to integrate data on numbers of CHD patients, treatment uptake, treatment effectiveness, population risk factor trends, and median survival among US adults. There were 308,900 fewer CHD deaths in 2000 among Americans aged 25-84 years than if 1980 mortality rates had applied. These 308,900 fewer deaths represented approximately 3,147,800 life-years gained (sensitivity analysis range, 2,448,900-3,744,900). Treatments for patients accounted for approximately 1,092,400 (751,700-1,387,000) life-years gained, whereas changes in the prevalence of population risk factors accounted for a gain of 2,055,500 (1,697,200-2,346,300) life-years. However, the 2,770,500 life-years gained through decreased levels of smoking, cholesterol, blood pressure, and physical inactivity were diminished by a loss of 715,000 life-years attributable to increased rates of obesity and diabetes. Therefore, modest reductions in the prevalence of several major cardiovascular disease risk factors accounted for more than twice as many life-years gained as did treatments. Unfortunately, these gains were partially offset by substantial increases in obesity and diabetes. PMID- 19541857 TI - Alcohol and risk of breast cancer by histologic type and hormone receptor status in postmenopausal women: the NIH-AARP Diet and Health Study. AB - Little is known about the association between alcohol and breast cancer by different tumor characteristics. The study consisted of 184,418 postmenopausal women aged 50-71 years in the National Institutes of Health-AARP Diet and Health Study (1995-2003). Alcohol use, diet, and potential risk factors for cancer were assessed with a mailed questionnaire at baseline. The relative risks and 95% confidence intervals were estimated by using Cox proportional hazards regression. Breast cancer cases and estrogen receptor and progesterone receptor status were identified through linkage to state cancer registries. During an average of 7 years of follow-up, 5,461 breast cancer cases were identified. Alcohol was significantly positively associated with total breast cancer: Even a moderate amount of alcohol (>10 g/day) significantly increased breast cancer risk. In a comparison of >35 g versus 0 g/day, the multivariate relative risks were 1.35 (95% confidence interval (CI): 1.17, 1.56) for total breast cancer, 1.46 (95% CI: 1.22, 1.75) for ductal tumors, and 1.52 (95% CI: 0.95, 2.44) for lobular tumors. The multivariate relative risks for estrogen receptor-positive/progesterone receptor-positive, estrogen receptor-positive/progesterone receptor-negative, and estrogen receptor-negative/progesterone receptor-negative tumors were 1.46 (95% CI: 1.12, 1.91) for >35 g versus 0 g/day, 1.13 (95% CI: 0.73, 1.77) for >20 g versus 0 g/day, and 1.21 (95% CI: 0.79, 1.84) for >20 g versus 0 g/day, respectively. Moderate consumption of alcohol was associated with breast cancer, specifically hormone receptor-positive tumors. PMID- 19541858 TI - Contact with beach sand among beachgoers and risk of illness. AB - Recent studies of beach sand fecal contamination have triggered interest among scientists and in the media. Although evidence shows that beach sand can harbor high concentrations of fecal indicator organisms, as well as fecal pathogens, illness risk associated with beach sand contact is not well understood. Beach visitors at 7 US beaches were enrolled in the National Epidemiological and Environmental Assessment of Recreational Water (NEEAR) Study during 2003-2005 and 2007 and asked about sand contact on the day of their visit to the beach (digging in the sand, body buried in the sand). Then, 10-12 days after their visit, participants were telephoned to answer questions about any health symptoms experienced since the visit. The authors completed 27,365 interviews. Digging in the sand was positively associated with gastrointestinal illness (adjusted incidence proportion ratio (aIPR) = 1.13, 95% confidence interval (CI): 1.02, 1.25) and diarrhea (aIPR = 1.20, 95% CI: 1.05, 1.36). The association was stronger between those buried in the sand and gastrointestinal illness (aIPR = 1.23, 95% CI: 1.05, 1.43) and diarrhea (aIPR = 1.24, 95% CI: 1.01, 1.52). Nonenteric illnesses did not show a consistent association with sand contact activities. Sand contact activities were associated with enteric illness at beach sites. Variation in beach-specific results suggests that site-specific factors may be important in the risk of illness following sand exposure. PMID- 19541859 TI - Characterization of the hCG variants recognized by different hCG immunoassays: an important step toward standardization of hCG measurements. PMID- 19541860 TI - Distribution and correlates of midregional proadrenomedullin in the general population. PMID- 19541861 TI - Performance of first-trimester screening between gestational weeks 7 and 13. AB - BACKGROUND: Screening for fetal chromosome abnormalities in the first trimester includes analysis of the serological markers pregnancy-associated plasma protein A (PAPP-A) and free beta human choriogonadotropin (free beta hCG). The blood sample is traditionally taken around week 12 of gestation, but the performance of earlier blood sampling is not well documented. METHODS: We studied 44,537 singleton pregnancies. Complete first-trimester screening took place between November 2003 and March 2009, and blood samples were taken between 7 weeks + 5 days and 13 weeks + 6 days. RESULTS: Of 120 cases of trisomy 21, 108 were diagnosed in the first-trimester screening (detection rate 90%). When the blood sample was taken before gestational week 10, the detection rate of trisomy 21 was 97% (70 of 72), whereas 80% were detected (38 of 48) after week 10 (chi(2) = 0.0035). For trisomy 18, trisomy 13, and triploidy, 65% (13 of 20) were detected before gestational week 10, and 73% (11 of 15) after (not significant). All 6 cases of triploidy before and after gestational week 10 were detected. The screen positive rate and the maternal age were similar before and after week 10 of gestation. CONCLUSIONS: Screening for fetal aneuploidy can be performed with good results with the blood sample taken as early as week 7 of gestation. Blood samples taken before gestational week 10 showed a high detection rate of fetal trisomy 21, with no difference in the detection of fetal trisomy 18, trisomy 13, or triploidy. PMID- 19541862 TI - Understanding resistance to tamoxifen in hormone receptor-positive breast cancer. PMID- 19541863 TI - Multiplex detection of 60 hepatitis B virus variants by maldi-tof mass spectrometry. AB - BACKGROUND: Variations in the hepatitis B virus (HBV) genome may develop spontaneously or under selective pressure from antiviral therapy. Such variations may confer drug resistance or affect virus replication capacity, resulting in failure of antiviral therapy. METHODS: A duplex PCR was used to amplify the region of the reverse transcriptase gene, the precore promoter, and the basal core promoter of the HBV genome. Four multiplex primer-extension reactions were used to interrogate 60 frequently observed HBV variants during antiviral therapy. Automated MALDI-TOF mass spectrometry (MS) was used for mutation detection. Capillary sequencing was used to confirm the MS results. RESULTS: The limit of quantification was 1000 HBV copies/mL for multiplex detection of HBV variants. Fifty-three variants (88.3%) were analyzed successfully in at least 90% of the sera from 88 treatment-naive patients and 80 patients with virologic breakthrough. MS was able to detect twice as many minor variants as direct sequencing while achieving close to full automation. MS and direct sequencing showed only 0.1% discordance in variant calls. CONCLUSIONS: This platform based on multiplex primer extension and MALDI-TOF MS was able to detect 60 HBV variants in 4 multiplex reactions with accuracy and low detection limits. PMID- 19541864 TI - Differences in recognition of the 1st WHO international reference reagents for hCG-related isoforms by diagnostic immunoassays for human chorionic gonadotropin. AB - BACKGROUND: The 1st WHO International Reference Reagents (IRRs) for 6 human chorionic gonadotropin (hCG)-related molecular variants, highly purified and calibrated in substance concentrations by the IFCC Working Group for hCG, permit experimental elucidation of what commercially available hCG methods measure in molar terms and enable assessment of their fitness for clinical purposes. METHODS: Pools containing known amounts of the IRRs spiked into normal human serum were issued to participants through the UK National External Quality Assessment Service for hCG for a period of 7 years. Among 16 assays used, 4 recognized only hCG, whereas 6 recognized hCG and its free beta-subunit (hCGbeta), and 6 recognized hCG, hCGbeta, and the beta core fragment. RESULTS: Differences in calibration of current hCG assays are moderate. Mean recovery of the current International Standard (IS), hCG IS 75/589, was 107% (range 93% to 126%), whereas that of the IRR 99/688 for hCG was 139% (range 109%-164%). Between method variation for the latter (CV 12.3%) was also greater than for IS 75/589 (CV 8.8%). Recognition of hCGbeta varied markedly (CV 37%). Most assays overestimated it, but 2 RIAs produced results that were slight underestimations. Recognition of the beta core fragment was even more variable (CV 57%) and was closest to equimolarity for the RIAs. CONCLUSIONS: Assays for hCG show considerable variation in their recognition of various forms of hCG, and this variability is the most important cause of method-related differences in hCG results in serum and an even more important cause of method-related differences in urine measurements. Equimolar recognition of the major hCG isoforms is essential if between-method comparability for hCG is to be improved. PMID- 19541866 TI - CYP2D6 genotyping for functional-gene dosage analysis by allele copy number detection. AB - BACKGROUND: Cytochrome P450 2D6 (CYP2D6), one of the most important drug metabolizing enzymes, has been reported to possess variation in the encoding CYP2D6 gene (cytochrome P450, family 2, subfamily D, polypeptide 6) that affects enzymatic activity. For the pharmacogenetic study of CYP2D6, accurate measurement of the dosage of the functional gene is essential; however, current genotyping techniques are insufficient because of their inability to provide the exact copy number of functional CYP2D6 genes. METHODS: We developed 3 quantitative real-time PCR (qPCR) assays for estimating the total copy number of the CYP2D6 gene, as well as 24-multiplex PCR-based real-time Invader assays (mPCR-RETINAs) for estimating the allele ratio at each variation locus. After determining the allele copy number at each locus, we estimated the frequencies of CYP2D6 alleles in a population and the diplotype in each individual by a CNVphaser (copy number variation phaser). The qPCR assays and RETINAs used for HapMap Japanese and Chinese samples were applied to 455 Japanese individuals. RESULTS: Forty-two individuals (9.2%) had one CYP2D6 gene copy, 207 (45.5%) had 2 copies, 161 (35.4%) had 3 copies, 40 (8.8%) had 4 copies, and 5 (1.1%) had 5 copies of the CYP2D6 gene. We found 16 different CYP2D6 alleles, with frequencies similar to those described in previous reports. In the diplotype analysis, we observed that CYP2D6*1/*1 and *1/*10-*36 were the most common diplotypes (approximately 20%) in our population. CONCLUSIONS: Our method is the first to determine the exact number of functional CYP2D6 gene copies. We believe our method will facilitate and accelerate the detailed pharmacogenetic analysis of CYP2D6. PMID- 19541865 TI - Asymmetric dimethylarginine reference intervals determined with liquid chromatography-tandem mass spectrometry: results from the Framingham offspring cohort. AB - BACKGROUND: Accumulating evidence links higher circulating asymmetric dimethylarginine (ADMA) to greater risk of cardiovascular disease (CVD). Relatively small differences in ADMA concentrations between healthy individuals and those with disease underscore the need to formulate reference intervals that may aid risk stratification of individuals. METHODS: We formulated reference intervals for plasma ADMA concentrations using a community-based reference sample from the Framingham Offspring Study consisting of 1126 nonsmoking individuals [mean (SD) age 56 (9) years; 60% women] who were free of clinical CVD, hypertension, diabetes, and obesity and who attended a routine examination at which ADMA was assayed. ADMA concentrations were determined using a validated tandem mass spectrometry-liquid chromatography assay. RESULTS: In the study sample, the mean ADMA concentration was 0.52 (0.11) micromol/L, and the reference limits were 0.311 and 0.732 (2.5th and 97.5th percentile). The sex-specific reference limits were 0.310 and 0.745 in men and 0.313 and 0.721 micromol/L in women. In multivariable regression analysis, ADMA plasma concentrations were positively correlated with age and total plasma homocysteine (both P < 0.001). CONCLUSIONS: Reference limits calculated for circulating ADMA in our large community-based healthy reference sample confirm the previous observation of a relatively narrow distribution of concentrations. This suggests a tight physiological control of ADMA plasma concentrations, presumably by dimethylarginine dimethylaminohydrolase (DDAH) metabolism of ADMA. PMID- 19541868 TI - Preanalytical stability of 25(OH)-vitamin D3 in human blood or serum at room temperature: solid as a rock. PMID- 19541869 TI - Opportunity value. PMID- 19541867 TI - Comparative analysis of PCR-based biomarker assay methods for colorectal polyp detection from fecal DNA. AB - BACKGROUND: Aberrantly methylated genes are promising biomarkers for the detection of colon adenomas and colorectal cancers (CRCs). The optimal assay type and specific methylated genes for these assays remain to be determined. METHODS: We used genomewide microarray-based assays to identify methylated genes as candidate biomarkers for colon neoplasms. The frequency of aberrant methylation of these genes in primary tumors was assessed with methylation-specific PCR (MSP). The limits of detection and specificities for different types of PCR-based assays were then assessed with the most promising genes identified in this screen. Finally, we assessed the best-performing MSP assay as an early-detection marker using fecal DNA samples. RESULTS: ITGA4 [integrin, alpha 4 (antigen CD49D, alpha 4 subunit of VLA-4 receptor)] was identified as a novel gene frequently methylated in CRC. Methylated ITGA4 is present in 75% of colon adenomas (n = 36) and 92% of colon adenocarcinomas (n = 75). Comparison of end point MSP, end point MSP with clamped primers, and quantitative fluorescent MSP (qMSP) approaches revealed that both types of end point MSP assays could routinely detect as little as 70 pg DNA, whereas the qMSP assay could routinely detect as little as 7 pg. A fecal DNA qMSP assay for methylated ITGA4 can detect 69% of individuals with colon adenomas (n = 13) with a diagnostic specificity of 79% (n = 28). CONCLUSIONS: Methylated ITGA4 is a promising marker gene for the early detection of colonic neoplasms. qMSP has the lowest limit of detection of the MSP assay types tested, and a qMSP assay that detects methylated ITGA4 has potential as an early-detection assay for colon neoplasms. PMID- 19541870 TI - Paying for better outcomes - the English way. PMID- 19541871 TI - Sources of variation in the costs of health care for asthma patients in Australia. AB - OBJECTIVES: Individuals with chronic conditions, such as asthma, on average incur high health care costs, though good control can reduce costs and improve health outcomes. However, there may be substantial variation between patients in their use of services and therefore costs. Our objective was to investigate the sources of such variation in health system and out-of-pocket costs for people with asthma. METHODS: A longitudinal observational study of 252 people with asthma in New South Wales, Australia, followed for three years, using six-monthly postal surveys and individual administrative data. Factors associated with costs were investigated using generalized linear mixed models. RESULTS: There was substantial variability in costs between individuals but relatively little within person change over time for the majority. Costs to the health system and out-of pocket costs were higher with increasing asthma-related health problems and increasing age. Health system costs were less for patients living outside the state capital (Sydney) and for those in the middle income group relative to high and low income groups. CONCLUSIONS: Those with poorly-controlled asthma and the elderly require more carefully targeted strategies to improve their health and ensure appropriate use of resources. Access to appropriate services for those living outside of major cities should be improved. Co-payments for the middle income groups and those living outside major cities should be reduced to improve equity in the use of services. PMID- 19541872 TI - Delivering health care through community pharmacies: are working conditions deterring female pharmacists' participation? AB - OBJECTIVE: Recent UK government policy has placed community pharmacists at the frontline of health care delivery in order to improve patient access. Community pharmacy has been beset by recruitment and retention problems which potentially threaten health service delivery. This is largely a consequence of an increased demand for pharmacists. Additionally, the proportion of female pharmacists in the profession has risen. Consequently, interrupted career patterns and part-time working practices have increased, shrinking the pool of available workers. This study aimed to examine the importance of factors influencing female community pharmacists' work patterns. METHOD: Q methodology was used in a sample of 40 female UK-based community pharmacists. RESULTS: Nine distinct factors emerged from a factor analysis of Q sorts: fulfilled pharmacists; family first or pharmacy shelved; low stress altruist; permanent part-time employees; focused on free time and finances; pressurized modernizers; wandering wage slaves; overloaded and under resourced for the new contract; and pin money part-timers. Female community pharmacists often worked below their potential and part-time at a practitioner level in response to a combination of domestic commitments and intensifying work place pressures. CONCLUSIONS: Family-friendly flexible work environments, adequate staffing levels and improved management support, might be more effective in increasing workforce participation than enhanced salary levels in this group of workers. PMID- 19541873 TI - Patient and hospital characteristics associated with claims and compensations for patient injuries in coronary artery bypass grafting in Finland. AB - OBJECTIVES: To analyse the association between individual patients' risk factors and rates of claims and compensations for patient injuries in an insurance scheme in which proof of negligence is not required. And to explore whether either hospital productivity or volume of procedures is related to claims and compensation rates. METHODS: A two-step sequential logistic regression was applied on data collected from administrative registers. It included 17,834 patients who had undergone coronary artery bypass grafting at public hospitals in Finland between 1998 and 2002. The main outcome measure was the odds of claiming and receiving compensation. RESULTS: Men were less likely to claim compensation (odds ratio [OR] 0.66; 95% confidence interval 0.54-0.81), but among those having claimed were more likely to receive compensation (OR 2.08; 1.15-3.75) than women. Patients with a co-morbidity were more likely to claim (OR 1.29; 1.06-1.57), but among those having claimed were less likely to receive compensation (OR 0.52; 0.31-0.86) than those without a co-morbidity. Advanced age reduced the probability of claiming (OR 0.71; 0.52-0.96). CONCLUSIONS: Although high-risk patients file a claim more frequently than low-risk patients, the latter have a higher probability of getting their claims accepted and receiving compensation. This risk pattern is probably a reflection of compensation practices related to patient injuries involving an infection. PMID- 19541874 TI - Developing a framework for transferring knowledge into action: a thematic analysis of the literature. AB - OBJECTIVES: Although there is widespread agreement about the importance of transferring knowledge into action, we still lack high quality information about what works, in which settings and with whom. While there are a large number of models and theories for knowledge transfer interventions, the majority are untested, meaning that their applicability and relevance is largely unknown. This paper describes the development of a conceptual framework of translating knowledge into action and discusses how it can be used for developing a useful model of the knowledge transfer process. METHODS: A narrative review of the knowledge transfer literature identified 28 different models which explained all or part of the knowledge transfer process. The models were subjected to a thematic analysis to identify individual components and the types of processes used when transferring knowledge into action. The results were used to build a conceptual framework of the process. RESULTS: Five common components of the knowledge transfer process were identified: problem identification and communication; knowledge/research development and selection; analysis of context; knowledge transfer activities or interventions; and knowledge/research utilization. We also identified three types of knowledge transfer processes: a linear process; a cyclical process; and a dynamic multidirectional process. From these results a conceptual framework of knowledge transfer was developed. The framework illustrates the five common components of the knowledge transfer process and shows that they are connected via a complex, multidirectional set of interactions. As such the framework allows for the individual components to occur simultaneously or in any given order and to occur more than once during the knowledge transfer process. CONCLUSION: Our framework provides a foundation for gathering evidence from case studies of knowledge transfer interventions. We propose that future empirical work is designed to test and refine the relevance, importance and applicability of each of the components in order to build a more useful model of knowledge transfer which can serve as a practical checklist for planning or evaluating knowledge transfer activities. PMID- 19541875 TI - Tackling climate change close to home: mobile breast screening as a model. AB - OBJECTIVE: Health services contribute significantly to carbon dioxide (CO(2)) emissions and, while services in the UK are beginning to address this, the focus has been on reducing energy consumption rather than road transport, a major component of emissions. We aimed to compare the distances travelled by patients attending mobile breast screening clinics compared to the distance they would need to travel if screening services were centralized. METHODS: Anonymized postcode records were analysed to determine driving distances potentially saved through attendance at 20 mobile breast screening clinics rather than at two centralized locations. Based on assumptions for the typical car used, the CO(2) emissions were calculated for the current case of decentralized service through mobile clinics compared to a hypothetical case where only centralized services are available over one complete three-year cycle of breast screening invitations. RESULTS: The availability of mobile breast screening clinics for the 60,675 women who underwent screening over a three-year cycle led to a return journey distance savings of 1,429,908 km. Taking into account the CO(2) emissions of the tractor unit used for moving the mobile clinics around, this equates to approximately 75 tonnes of CO(2) saved in any one year. CONCLUSIONS: Decentralizing health care delivery can potentially provide substantial reductions in emissions at the same time as improving the patient experience. Thus, the 'care close to home' agenda can simultaneously improve health outcomes and the environment. PMID- 19541876 TI - Reducing waiting times for hospital treatment: lessons from the English NHS. AB - In recent years, the English NHS has achieved substantial reductions in waiting times for hospital treatment. This paper considers first whether the data used by the Government provide an accurate description of changes in waiting times and identifies some of the limitations of the measures used. It then attempts to identify how reductions have been achieved. It argues that some features of central government policy have been important - such as the use of targets - others, such as the introduction of new private sector capacity have not. It also shows that changes at local level have been critical to achieving the recorded improvements, but the precise impact of these is hard to identify. PMID- 19541877 TI - What is the relationship between patients' and clinicians' reports of the outcomes of elective surgery? AB - OBJECTIVES: To identify studies in which patients' and clinicians' reports of health status and complications of one of four elective operations - hip and knee replacement, varicose vein surgery and groin hernia repair - are reported, and to describe the associations that have been reported between clinicians' and patients' reports. METHODS: Systematic search of several bibliographic databases and review of citations of articles meeting inclusion criteria. A narrative summary of the findings was conducted. RESULTS: Most of the 62 studies of health status identified were for hip (23) or knee (33) disease. The literature on complications was even more limited with 12 studies of surgical site infection, one for urinary tract infection and none for lower respiratory tract infections. Procedure-specific complications were restricted to one for arthroplasties and three for hernia repair. Despite considerable variation in the findings of studies, some clear patterns emerge, albeit they are largely based on arthroplasty. Patients' and clinicians' views of health status generally correlate moderately (0.5-0.6) when both are reporting on the same dimension of health status. Inevitably this is confined to disability, though patients' and clinicians' reports of symptoms are also moderately correlated. In contrast, comparisons of different dimensions, such as patients' reports of disability and clinicians' reports of impairment, result in poor correlation (0.3). There is huge variation in the way postoperative complications are measured which limits the extent to which an overview can be undertaken. Despite that, moderate to strong correlations have been reported between patients' and clinicians' views of complications. CONCLUSIONS: Patients' views of their level of disability reflect clinicians' views and can be relied upon to assess this dimension of health status. In addition, patients are the 'gold standard' judges of symptoms and quality of life. Given these findings, clinicians, provider managers, commissioners and politicians can be confident that patients' reports provide an accurate indication of the outcome of elective surgery. PMID- 19541878 TI - Measuring the quality of medical care. PMID- 19541879 TI - UK chiropractic: regulated but unruly. AB - Since 1994 chiropractic has been regulated by statute in the UK. Despite this air of respectability, a range of important problems continue to bedevil this profession. Professional organizations of chiropractic and their members make numerous claims which are not supported by sound evidence. Many chiropractors adhere to concepts which fly in the face of science and most seem to regularly violate important principles of ethical behaviour. The advice chiropractors give to their clients is often dangerously misleading. If chiropractic in the UK is to grow into an established health care profession, the General Chiropractic Council and its members should comply with the accepted standards of today's health care. PMID- 19541880 TI - In praise of chiropractic. AB - The rationales used in chiropractic are largely in keeping with current evidence based guidance. When listening to debates for and against the scientific basis of chiropractic, it is important to be aware of the selective use of evidence on both sides and of the limitations of logical positivistic arguments when it comes to health care areas that have a substantial psychosocial component. PMID- 19541881 TI - Comparison of queen-specific gene expression in related lower termite species. AB - The molecular mechanisms regulating caste determination and reproductive division of labor, the hallmarks of insect societies, are poorly defined. The identification of key genes involved in these developmentally important processes will be essential to gain a better understanding of the mechanisms controlling one of the most impressive examples of polyphenism, the caste structure of eusocial species. Here, we applied representational difference analysis of cDNAs, to study differential gene expression between queens (female neotenics) and workers in the dry wood termite Cryptotermes cynocephalus and identified 13 genes that were highly expressed in queens. In addition, we partially cloned several homologous genes of the related termite species Cryptotermes secundus and compared the expression profiles of 10 homologous genes. In most cases, the preferential expression in female neotenics was not conserved between species, despite the close phylogenetic relationship of both Cryptotermes species. It is possible that these genes are associated with known species-specific differences in caste development modes. Only three genes (Neofem1, 2, and 3) showed a conserved and highly preferential expression in female neotenics, suggesting that their products may play important roles in female reproductives, in particular in controlling caste determination and reproductive division of labor. PMID- 19541882 TI - Girls with social and/or attention deficits: a descriptive study of 100 clinic attenders. AB - OBJECTIVE: Examine clinical correlates and distinguishing features of autism spectrum disorders (ASD), ADHD, and tic disorders in girls referred for social impairment, attention/academic deficits, and/or tics. METHOD: One hundred 3- to 18-year-old girls referred for social impairment and attention symptoms were assessed in detail. Sixty of these girls, 7 to 16 years of age (IQ >or= 80) were compared with age-matched girls (IQ >or= 80) from the community. RESULTS: Main diagnoses of ASD, ADHD, tic disorders, and "other psychiatric disorder" were made in 46, 46, 3, and 5, respectively, of the referred girls. The ASD and ADHD groups (mean age at diagnosis 8.8 and 13.0 years, respectively) had the same types and high rates of psychiatric comorbidity. Girls with ASD had more problems with global functioning and adaptive levels of daily living skills than girls with ADHD. Differences between these girls referred for investigation and the community sample of girls were very considerable across a range of factors. CONCLUSIONS: Girls referred for social and/or attention deficits usually meet diagnostic criteria for either ASD or ADHD. They have severe psychiatric comorbidities and low global levels of functioning. PMID- 19541883 TI - How to contain generalized HIV epidemics? A plea for better evidence to displace speculation. AB - In the worst generalized HIV epidemics in East and Southern Africa, from one quarter to three-quarters of women aged 15 years can expect to be living with HIV or to have died with AIDS by age 40 years. This disaster continues in the face of massive HIV prevention programmes based on current inexact knowledge of HIV transmission pathways and risks. To stop this disaster, both the public and public health experts need better information about the specific factors that allow HIV to propagate so extensively in countries with generalized epidemics. This knowledge could be acquired by tracing HIV infections to their source - especially tracing HIV infections in women of all ages, and tracing unexplained HIV infections in children with HIV-negative mothers. PMID- 19541884 TI - A brief review of potential mechanisms of immune reconstitution inflammatory syndrome in HIV following antiretroviral therapy. AB - A subset of HIV-infected individuals who receive antiretroviral therapy (ART) develop a paradoxical pathological response that significantly increases morbidity and sometimes mortality. Following the induction of highly active ART, a rapid decline in the viral load results within weeks and coincides with a steep rise in the CD4(+) T-cell counts and immune hyperactivation. Although no mechanistic pathway has been elucidated for the development of immune reconstitution inflammatory syndrome (IRIS), it is thought that change in the nature of the immune response is a predominant factor in the development of reconstitution disease. In this article, we review the current state of knowledge in this field and provide a model for the development of IRIS. PMID- 19541885 TI - 2009 European (IUSTI/WHO) guideline on the diagnosis and treatment of gonorrhoea in adults. PMID- 19541886 TI - 2009 European guideline on the management of male non-gonococcal urethritis. PMID- 19541887 TI - Chlamydia trachomatis infection during pregnancy and the risk of preterm birth: a case-control study. AB - Our goal was to define the risks of preterm birth associated with Chlamydia trachomatis (CT) and other sexually transmitted infections (STIs) among pregnant women. We accessed clinical records from July 2005 to February 2008. The study population included all pregnant women who gave birth to a singleton newborn of at least 20 weeks' gestation, and who had antenatal care information. We estimated the impact of CT and other STI on the odds of preterm birth using logistic regression. Overall, 2127 women were included in this analysis. The prevalence of CT infection was 4.7%. CT diagnosis was not associated with preterm birth. In conclusion, this study did not find an association between CT and preterm birth. The lack of an association may be explained by early treatment. Future studies evaluating the timing of screening for STIs may help clarify whether pregnant women would benefit more from earlier screening. PMID- 19541888 TI - Targeting of HIV pretest discussion and risk-reduction interventions in a low prevalence setting. AB - Genitourinary medicine clinic attendees who were newly diagnosed as HIV antibody positive in a population with low overall HIV prevalence were compared with HIV ve control groups in a retrospective study. Demographics and clinical data from the clinic attendance at which the HIV test was performed were analysed. Of 25,627 HIV tests, 113 were positive. Seventy-eight percent had an identified risk factor for HIV and more than half had symptoms of, or were contacts of, a sexually transmitted infection (STI) or HIV. Only eight clients testing HIV+ve had attended for routine STI testing and only two of these had no identified risk factor. Groups shown to be at higher HIV risk included attendees with past history of STI, men-who-have-sex-with-men (MSM) and those requesting HIV test without an STI screen. MSM testing HIV-ve had high rates of HIV risk behaviour. PMID- 19541889 TI - Prevention of human immunodeficiency virus mother-to-child transmission in Israel. AB - The objective of the study was to investigate the HIV-mother-to-child transmission (MTCT) rate in Israel. This was a retrospective study of HIV infected pregnant women, mainly immigrants from Ethiopia, in six Israeli AIDS centres, in 2000-2005. Medical records of mothers and newborns were evaluated for HIV status, treatment and MTCT rates. Three hundred pregnancies of 241 HIV infected women, resulting in 304 live births, were studied. In 86/241(36%) women, HIV diagnosis was made during the current pregnancy or shortly after labour. Thirty others were diagnosed during previous pregnancies. Highly active antiretroviral therapy (HAART) was prescribed in 76% of pregnancies. The mean viral load before labour was 23,000 +/- 100,000 copies/mL with a mean CD4 of 406 +/- 223 (range 4-1277) cells/mm(3). Caesarian sections were preformed in 175/300 pregnancies (103/175 with viral load <1000 copies/mL). During labour, azidothymidine (AZT) was given to 80% and nevirapine to 8% of the women. Eighty eight percent of the neonates received AZT for six weeks. The overall HIV-MTCT rate was 3.6%. MTCT correlated significantly with delayed HIV diagnosis, low CD4, lack of HAART during pregnancy and lack of perinatal treatment. HIV treatment of mothers and their newborns throughout pregnancy, labour and perinatal period are crucial for effective prevention of MTCT, emphasizing the need for early HIV screening, diagnosis and treatment. PMID- 19541890 TI - HIV infection and associated risk factors among long-distance truck drivers travelling through Azerbaijan. AB - The aim of this study was to assess HIV prevalence and associated risk behaviours among international truck drivers (TDs) in Azerbaijan. The subjects signed consent and completed a questionnaire. Blood was tested using two rapid HIV tests: Determine and OraQuick. Genotyping was performed on 13 positives. Overall, 3763 TDs from 21 countries were enrolled. Fifty-eight (1.54%) were HIV-positive. Highest prevalence was among Russians (2.88%), Ukrainians (1.66%) and Azerbaijani (1.09%). On univariate analysis, highest prevalence (60%) was among injecting drug users (IDUs) compared with 0.4% among non-IDUs (P < 0.001). The prevalence in men who had sex with men (MSM) (42.9%) was high (P 500 cells/microL were anaemic, compared with 64% of those with CD4 counts <100 cells/microL (P < 0.001). In multivariate analysis, CD4 count <100 cells/microL (odds ratio [OR]:5.0, confidence interval [CI]:4.0-6.3), underweight body mass index (OR:4.8, CI:3.6-6.5), female gender (OR:3.1, CI:2.8-3.6) and tuberculosis (TB) (OR:1.6, CI:1.4-1.8) were significantly associated with anaemia. In this setting, management of anaemia should focus on antiretroviral therapy, nutritional supplementation and TB control. The high anaemia prevalence among patients meeting criteria for antiretroviral therapy highlights the need for increased access to non-zidovudine nucleoside reverse transcriptase inhibitors in developing countries. PMID- 19541894 TI - Fourteen years of surveillance of HIV-1 prevalence among pregnant women attending antenatal care clinics in western Uganda. AB - This study monitored long-term temporal trends in HIV-1 prevalence in antenatal clinic attendees living in western Uganda. Semi-annual data collection was done from 1991 to 2004. For each woman the following data were recorded: HIV-1 status, age, educational status, marital status, occupation and parity. The results show that the overall HIV-1 prevalence was 15.3% during the entire time period (urban 21.3%, semi-urban 12.7% and rural 7.1%). Between 1991 and 2004, we observed a gradual decline in the HIV-1 prevalence. The decline was most pronounced in urban women aged 15-19 years old and least pronounced in rural women aged 20-24 years. Women above 25 years of age did not show any decline in HIV-1 prevalence over time. The declining HIV-1 prevalence in the younger age groups (15-24 years) likely represents a declining risk for acquiring HIV infection as we have previously shown in the urban sub-sample of this data set. PMID- 19541895 TI - General practitioner involvement and patient outcomes in HIV management. AB - We undertook this study to try to determine whether disease outcomes were poorer in patients with HIV infection whose general practitioner (GP) was unaware of their status compared with those whose GP was aware. The notes of 375 HIV positive patients attending Edinburgh's genitourinary (GU) medicine clinic were reviewed. The GPs of 292 patients (78%) had been informed of their patient's HIV infection. Advancing disease was associated with disclosure of the status to GPs (P = 0.037) but no significant association was found between informing GPs and the viral load results of treated (P = 0.389) and untreated patients (P = 0.070). Twenty-three percent of patients had had one or more bacterial sexually transmitted infections (STIs) while receiving their HIV care at a GU medicine clinic. Patients diagnosed with an STI were less likely to disclose their HIV status to their GP (P < 0.0005). Non-disclosure of the HIV status to a GP may be a predictor of unsafe sexual practices. PMID- 19541896 TI - Why are HIV-positive mothers refusing to have their children screened for vertically transmitted HIV infection? AB - A previous audit showed that the HIV status of the majority (73%) of children of HIV-positive mothers attending a genitourinary clinic in the United Kingdom was unknown because mothers did not take up the offer of testing. The objectives of a re-audit were to establish the impact of the audit process on the uptake of testing and reasons for not taking up the offer of screening of offspring. One year after the previous audit, 13/92 (14%) of children not previously tested had their HIV status established. The reason for not testing was, in 43/52 (82%) mothers, the perception that a well child can not be infected with HIV. This was the only reason for not having their children tested in 16 mothers (31%). The next most common theme was fear of disclosure to others, in 29 (56%). The barriers for screening included fear of feeling guilty if the child was found to be positive. PMID- 19541897 TI - Sexual health of adults working in pornographic films. AB - We report the frequency of sexually transmitted infections (STIs) diagnosed in performers in the adult pornographic film industry. Over a 13 month period, 445 STI screens were performed in 115 patients, 56 women and 59 men. All reported unprotected sex during filming. Seventy-five percent (86) had at least one sexual partner outside work, and 90% used condoms inconsistently with them. Women worked exclusively with women (23%), men only (38%) or both genders (39%). Almost all men (97%) worked exclusively heterosexually. Thirty-eight percent (44/115) were diagnosed with 77 STIs, including non-specific urethritis (51), gonorrhoea (10), chlamydia (6) and genital warts (6). Gonorrhoea was found exclusively at the pharynx in three heterosexual men. There were no cases of HIV, syphilis, hepatitis B or hepatitis C. Monthly screening and certification is a working requirement for this population but STIs are common in an industry where unprotected sex is the norm. PMID- 19541898 TI - Mondor's disease of the penis associated with primary syphilis. AB - A case of penile Mondor's disease associated with syphilitic chancre is reported. The aetiology and management of Mondor's disease is also discussed. PMID- 19541899 TI - Breakage of arm of Multiload intrauterine device during removal. AB - We report a case of intrauterine device (IUD) removal five years after its insertion which was broken during the removal procedure. One of the arms of the IUD remained inside the uterus. We allowed three months for spontaneous expulsion. When this did not happen, we proceeded with a hysteroscopic removal. Because of its embebbed position in the myometrium this could not be removed during hysteroscopy and finally it was left in situ. The patient was monitored every three months for a period of 12 months. There was no problem. We report this case because doctors may find it necessary to remove the IUD in certain situations and be aware of such a breakage and various precautions to prevent this. PMID- 19541900 TI - HIV positive: who do they tell? PMID- 19541901 TI - An entomological surprise. PMID- 19541908 TI - Increased locomotor activity induced by heroin in mice: pharmacokinetic demonstration of heroin acting as a prodrug for the mediator 6-monoacetylmorphine in vivo. AB - We investigated the relative importance of heroin and its metabolites in eliciting a behavioral response in mice by studying the relationship between concentrations of heroin, 6-monoacetylmorphine (6MAM), and morphine in brain tissue and the effects on locomotor activity. Low doses (subcutaneous) of heroin (< or =5 micromol/kg) or 6MAM (< or =15 micromol/kg) made the mice run significantly more than mice given equimolar doses of morphine. There were no differences in the response between heroin and 6MAM, although we observed a shift to the left of the dose-response curve for the maximal response of heroin. The behavioral responses were abolished by pretreatment with 1 mg/kg naltrexone. Heroin was detected in brain tissue after injection, but the levels were low and its presence too short-lived to be responsible for the behavioral response observed. The concentration of 6MAM in brain tissue increased shortly after administration of both heroin and 6MAM and the concentration changes during the first hour roughly reflected the changes in locomotor activity. Both the maximal and the total concentration of 6MAM were higher after administration of heroin than after administration of 6MAM itself. The morphine concentration increased slowly after injection and could not explain the immediate behavioral response. In summary, the locomotor activity response after injection of heroin was mediated by 6MAM, which increased shortly after administration. Heroin acted as an effective prodrug. The concentration of morphine was too low to stimulate the immediate response observed but might have an effect on the later part of the heroin-induced behavioral response curve. PMID- 19541907 TI - Mechanism of differential cardiovascular response to propofol in Dahl salt sensitive, Brown Norway, and chromosome 13-substituted consomic rat strains: role of large conductance Ca2+ and voltage-activated potassium channels. AB - Cardiovascular sensitivity to general anesthetics is highly variable among individuals in both human and animal models, but little is known about the genetic determinants of drug response to anesthetics. Recently, we reported that propofol (2,6-diisopropylphenol) causes circulatory instability in Dahl salt sensitive SS/JRHsdMcwi (SS) rats but not in Brown Norway BN/NHsdMcwi (BN) rats and that these effects are related to genes on chromosome 13. Based on the hypothesis that propofol does target mesenteric circulation, we investigated propofol modulation of mesenteric arterial smooth muscle cells (MASMC) in SS and BN rats. The role of chromosome 13 was tested using SS-13(BN)/Mcwi and BN 13(SS)/Mcwi consomic strains with chromosome 13 substitution. Propofol (5 microM) produced a greater in situ hyperpolarization of MASMC membrane potential in SS than BN rats, and this effect was abrogated by iberiotoxin, a voltage-activated potassium (BK) channel blocker. In inside-out patches, the BK channel number, P(o), and apparent Ca(2+) sensitivity, and propofol sensitivity all were significantly greater in MASMC of SS rats. The density of whole-cell BK current was increased by propofol more in SS than BN myocytes. Immunolabeling confirmed higher expression of BK alpha subunit in MASMC of SS rats. Furthermore, the hyperpolarization produced by propofol, the BK channel properties, and propofol sensitivity were modified in MASMC of SS-13(BN)/Mcwi and BN-13(SS)/Mcwi strains toward the values observed in the background SS and BN strains. We conclude that differential function and expression of BK channels, resulting from genetic variation within chromosome 13, contribute to the enhanced propofol sensitivity in SS and BN-13(SS)/Mcwi versus BN and SS-13(BN)/Mcwi strains. PMID- 19541909 TI - Can the whole be less than the sum of its parts? Pathway analysis in genome-scale metabolic networks using elementary flux patterns. AB - Elementary modes represent a valuable concept in the analysis of metabolic reaction networks. However, they can only be computed in medium-size systems, preventing application to genome-scale metabolic models. In consequence, the analysis is usually constrained to a specific part of the known metabolism, and the remaining system is modeled using abstractions like exchange fluxes and external species. As we show by the analysis of a model of the central metabolism of Escherichia coli that has been previously analyzed using elementary modes, the choice of these abstractions heavily impacts the pathways that are detected, and the results are biased by the knowledge of the metabolic capabilities of the network by the user. In order to circumvent these problems, we introduce the concept of elementary flux patterns, which explicitly takes into account possible steady-state fluxes through a genome-scale metabolic network when analyzing pathways through a subsystem. By being similar to elementary mode analysis, our concept now allows for the application of many elementary-mode-based tools to genome-scale metabolic networks. We present an algorithm to compute elementary flux patterns and analyze a model of the tricarboxylic acid cycle and adjacent reactions in E. coli. Thus, we detect several pathways that can be used as alternative routes to some central metabolic pathways. Finally, we give an outlook on further applications like the computation of minimal media, the development of knockout strategies, and the analysis of combined genome-scale networks. PMID- 19541910 TI - Next-generation tag sequencing for cancer gene expression profiling. AB - We describe a new method, Tag-seq, which employs ultra high-throughput sequencing of 21 base pair cDNA tags for sensitive and cost-effective gene expression profiling. We compared Tag-seq data to LongSAGE data and observed improved representation of several classes of rare transcripts, including transcription factors, antisense transcripts, and intronic sequences, the latter possibly representing novel exons or genes. We observed increases in the diversity, abundance, and dynamic range of such rare transcripts and took advantage of the greater dynamic range of expression to identify, in cancers and normal libraries, altered expression ratios of alternative transcript isoforms. The strand-specific information of Tag-seq reads further allowed us to detect altered expression ratios of sense and antisense (S-AS) transcripts between cancer and normal libraries. S-AS transcripts were enriched in known cancer genes, while transcript isoforms were enriched in miRNA targeting sites. We found that transcript abundance had a stronger GC-bias in LongSAGE than Tag-seq, such that AT-rich tags were less abundant than GC-rich tags in LongSAGE. Tag-seq also performed better in gene discovery, identifying >98% of genes detected by LongSAGE and profiling a distinct subset of the transcriptome characterized by AT-rich genes, which was expressed at levels below those detectable by LongSAGE. Overall, Tag-seq is sensitive to rare transcripts, has less sequence composition bias relative to LongSAGE, and allows differential expression analysis for a greater range of transcripts, including transcripts encoding important regulatory molecules. PMID- 19541911 TI - Circos: an information aesthetic for comparative genomics. AB - We created a visualization tool called Circos to facilitate the identification and analysis of similarities and differences arising from comparisons of genomes. Our tool is effective in displaying variation in genome structure and, generally, any other kind of positional relationships between genomic intervals. Such data are routinely produced by sequence alignments, hybridization arrays, genome mapping, and genotyping studies. Circos uses a circular ideogram layout to facilitate the display of relationships between pairs of positions by the use of ribbons, which encode the position, size, and orientation of related genomic elements. Circos is capable of displaying data as scatter, line, and histogram plots, heat maps, tiles, connectors, and text. Bitmap or vector images can be created from GFF-style data inputs and hierarchical configuration files, which can be easily generated by automated tools, making Circos suitable for rapid deployment in data analysis and reporting pipelines. PMID- 19541912 TI - Profiling the T-cell receptor beta-chain repertoire by massively parallel sequencing. AB - T-cell receptor (TCR) genomic loci undergo somatic V(D)J recombination, plus the addition/subtraction of nontemplated bases at recombination junctions, in order to generate the repertoire of structurally diverse T cells necessary for antigen recognition. TCR beta subunits can be unambiguously identified by their hypervariable CDR3 (Complement Determining Region 3) sequence. This is the site of V(D)J recombination encoding the principal site of antigen contact. The complexity and dynamics of the T-cell repertoire remain unknown because the potential repertoire size has made conventional sequence analysis intractable. Here, we use 5'-RACE, Illumina sequencing, and a novel short read assembly strategy to sample CDR3(beta) diversity in human T lymphocytes from peripheral blood. Assembly of 40.5 million short reads identified 33,664 distinct TCR(beta) clonotypes and provides precise measurements of CDR3(beta) length diversity, usage of nontemplated bases, sequence convergence, and preferences for TRBV (T cell receptor beta variable gene) and TRBJ (T-cell receptor beta joining gene) gene usage and pairing. CDR3 length between conserved residues of TRBV and TRBJ ranged from 21 to 81 nucleotides (nt). TRBV gene usage ranged from 0.01% for TRBV17 to 24.6% for TRBV20-1. TRBJ gene usage ranged from 1.6% for TRBJ2-6 to 17.2% for TRBJ2-1. We identified 1573 examples of convergence where the same amino acid translation was specified by distinct CDR3(beta) nucleotide sequences. Direct sequence-based immunoprofiling will likely prove to be a useful tool for understanding repertoire dynamics in response to immune challenge, without a priori knowledge of antigen. PMID- 19541913 TI - Evidence-based gene predictions in plant genomes. AB - Automated evidence-based gene building is a rapid and cost-effective way to provide reliable gene annotations on newly sequenced genomes. One of the limitations of evidence-based gene builders, however, is their requirement for transcriptional evidence-known proteins, full-length cDNAs, or expressed sequence tags (ESTs)-in the species of interest. This limitation is of particular concern for plant genomes, where the rate of genome sequencing is greatly outpacing the rate of EST- and cDNA-sequencing projects. To overcome this limitation, we have developed an evidence-based gene build system (the Gramene pipeline) that can use transcriptional evidence across related species. The Gramene pipeline uses the Ensembl computing infrastructure with a novel data processing scheme. Using the previously annotated plant genomes, the dicot Arabidopsis thaliana and the monocot Oryza sativa, we show that the cross-species ESTs from within monocot or dicot class are a valuable source of evidence for gene predictions. We also find that, using only EST and cross-species evidence, the Gramene pipeline can generate a plant gene set that is comparable in quality to the human genes based on known proteins and full-length cDNAs. We compare the Gramene pipeline to several widely used ab initio gene prediction programs in rice; this comparison shows the pipeline performs favorably at both the gene and exon levels with cross species gene products only. We discuss the results of testing the pipeline on a 22-Mb region of the newly sequenced maize genome and discuss potential application of the pipeline to other genomes. PMID- 19541914 TI - Abundant primary piRNAs, endo-siRNAs, and microRNAs in a Drosophila ovary cell line. AB - Piwi proteins, a subclass of Argonaute-family proteins, carry approximately 24-30 nt Piwi-interacting RNAs (piRNAs) that mediate gonadal defense against transposable elements (TEs). We analyzed the Drosophila ovary somatic sheet (OSS) cell line and found that it expresses miRNAs, endogenous small interfering RNAs (endo-siRNAs), and piRNAs in abundance. In contrast to intact gonads, which contain mixtures of germline and somatic cell types that express different Piwi class proteins, OSS cells are a homogenous somatic cell population that expresses only PIWI and primary piRNAs. Detailed examination of its TE-derived piRNAs and endo-siRNAs revealed aspects of TE defense that do not rely upon ping-pong amplification. In particular, we provide evidence that a subset of piRNA master clusters, including flamenco, are specifically expressed in OSS and ovarian follicle cells. These data indicate that the restriction of certain TEs in somatic gonadal cells is largely mediated by a primary piRNA pathway. PMID- 19541915 TI - Genome-wide comparisons of variation in linkage disequilibrium. AB - Current genome-wide surveys of common diseases and complex traits fundamentally aim to detect indirect associations where the single nucleotide polymorphisms (SNPs) carrying the association signals are not biologically active but are in linkage disequilibrium (LD) with some unknown functional polymorphisms. Reproducing any novel discoveries from these genome-wide scans in independent studies is now a prerequisite for the putative findings to be accepted. Significant differences in patterns of LD between populations can affect the portability of phenotypic associations when the replication effort or meta analyses are attempted in populations that are distinct from the original population in which the genome-wide study is performed. Here, we introduce a novel method for genome-wide analyses of LD variations between populations that allow the identification of candidate regions with different patterns of LD. The evidence of LD variation provided by the introduced method correlated with the degree of differences in the frequencies of the most common haplotype across the populations. Identified regions also resulted in greater variation in the success of replication attempts compared with random regions in the genome. A separate permutation strategy introduced for assessing LD variation in the absence of genome-wide data also correctly identified the expected variation in LD patterns in two well-established regions undergoing strong population-specific evolutionary pressure. Importantly, this method addresses whether a failure to reproduce a disease association in a disparate population is due to underlying differences in LD structure with an unknown functional polymorphism, which is vital in the current climate of replicating and fine-mapping established findings from genome-wide association studies. PMID- 19541918 TI - Best conventional therapy versus modular impairment-oriented training for arm paresis after stroke: a single-blind, multicenter randomized controlled trial. AB - BACKGROUND: The study investigated whether passive splinting or active motor training as either individualized best conventional therapy or as standardized impairment-oriented training (IOT) would be superior in promoting motor recovery in subacute stroke patients with mildly or severely paretic arms. METHODS: A total of 148 anterior circulation ischemic stroke patients were randomly assigned to 45 minutes of additional daily arm therapy over 3 to 4 weeks as either (a) passive therapy with inflatable splints or active arm motor therapy as either (b) individualized best conventional therapy (CONV) or (c) standardized IOT, that is Arm BASIS training for severe paresis or Arm Ability training for mild paresis. MAIN OUTCOME MEASURES: included the following: Fugl-Meyer arm motor score (severely paretic arms) and the TEMPA time scores (mildly affected arms). Pre post (immediate effects) and pre-4 weeks follow-up analyses (long-term effects) were performed. RESULTS: Overall improvements were documented (mean baseline and change scores efficacy: Fugl-Meyer, arm motor scores, 24.4, +9.1 points; TEMPA, 119, -26.6 seconds; P<.0001), but with no differential effects between splint therapy and the combined active motor rehabilitation groups. Both efficacy and effectiveness analyses indicated, however, bigger immediate motor improvements after IOT as compared with best conventional therapy (Fugl-Meyer, arm motor scores: IOT +12.3, CONV +9.2 points; TEMPA: IOT -31.1 seconds, CONV -20.5 seconds; P=.0363); for mildly affected patients long-term effects could also be substantiated. CONCLUSIONS: Specificity of active training seemed more important for motor recovery than intensity (therapy time). The comprehensive modular IOT approach promoted motor recovery in patients with either severe or mild arm paresis. PMID- 19541916 TI - Aerobic exercise improves cognition and motor function poststroke. AB - BACKGROUND: Cognitive deficits impede stroke recovery. Aerobic exercise (AEX) improves cognitive executive function (EF) processes in healthy individuals, although the learning benefits after stroke are unknown. OBJECTIVE: To understand AEX-induced improvements in EF, motor learning, and mobility poststroke. METHODS: Following cardiorespiratory testing, 38 chronic stroke survivors were randomized to 2 different groups that exercised 3 times a week (45-minute sessions) for 8 weeks. The AEX group (n = 19; 9 women; 10 men; 64.10 +/- 12.30 years) performed progressive resistive stationary bicycle training at 70% maximal heart rate, whereas the Stretching Exercise (SE) group (n = 19; 12 women; 7 men; 58.96 +/- 14.68 years) performed stretches at home. Between-group comparisons were performed on the change in performance at "Post" and "Retention" (8 weeks later) for neuropsychological and motor function measures. RESULTS: VO(2)max significantly improved at Post with AEX (P = .04). AEX also improved motor learning in the less-affected hand, with large effect sizes (Cohen's d calculation). Specifically, AEX significantly improved information processing speed on the serial reaction time task (SRTT; ie, "procedural motor learning") compared with the SE group at Post (P = .024), but not at Retention. Also, at Post (P = .038), AEX significantly improved predictive force accuracy for a precision grip task requiring attention and conditional motor learning of visual cues. Ambulation and sit-to-stand transfers were significantly faster in the AEX group at Post (P = .038), with balance control significantly improved at Retention (P = .041). EF measurements were not significantly different for the AEX group. CONCLUSION: AEX improved mobility and selected cognitive domains related to motor learning, which enhances sensorimotor control after stroke. PMID- 19541917 TI - Multicenter randomized trial of robot-assisted rehabilitation for chronic stroke: methods and entry characteristics for VA ROBOTICS. AB - BACKGROUND: Chronic upper extremity impairment due to stroke has significant medical, psychosocial, and financial consequences, but few studies have examined the effectiveness of rehabilitation therapy during the chronic stroke period. OBJECTIVE: . To test the safety and efficacy of the MIT-Manus robotic device for chronic upper extremity impairment following stroke. METHODS: . The VA Cooperative Studies Program initiated a multicenter, randomized, controlled trial in November 2006 (VA ROBOTICS). Participants with upper extremity impairment >/=6 months poststroke were randomized to robot-assisted therapy (RT), intensive comparison therapy (ICT), or usual care (UC). RT and ICT consisted of three 1 hour treatment sessions per week for 12 weeks. The primary outcome was change in the Fugl-Meyer Assessment upper extremity motor function score at 12 weeks relative to baseline. Secondary outcomes included the Wolf Motor Function Test and the Stroke Impact Scale. RESULTS: . A total of 127 participants were randomized: 49 to RT, 50 to ICT, and 28 to UC. The majority of participants were male (96%), with a mean age of 65 years. The primary stroke type was ischemic (85%), and 58% of strokes occurred in the anterior circulation. Twenty percent of the participants reported a stroke in addition to their index stroke. The average time from the index stroke to enrollment was 56 months (range, 6 months to 24 years). The mean Fugl-Meyer score at entry was 18.9. CONCLUSIONS: . VA ROBOTICS demonstrates the feasibility of conducting multicenter clinical trials to rigorously test new rehabilitative devices before their introduction to clinical practice. The results are expected in early 2010. PMID- 19541919 TI - Collaborative models for translational neuroscience and rehabilitation research. AB - Little formal research has been conducted on strategies to structure basic, preclinical, and clinical research to increase the likelihood of discovering efficacious interventions for patients with neurological diseases. How academic research is organized and funded by government agencies and foundations seems likely to affect the quality and rate of production of valued therapeutic agents. Few models for translational biomedical research, however, have been defined and no strategies have been compared. Given the narrow width of expertise and laboratory capacity of individual investigators, the complexity of identifying and manipulating mechanisms of disease components over time, and the demand for solutions from society, our continued reliance on funding therapeutic discovery through standalone investigators and projects seems counterproductive. Models are described for funding collaborations of basic and clinical scientists to work in iterative, adaptable, cross-disciplinary interactions around key progress limiting questions. Problem-oriented collaborations require leadership, incentives, trust, ongoing assessment, and an efficient infrastructure that overcomes barriers. These models are as testable as the hypotheses that drive scientific research. PMID- 19541920 TI - Direct-to-consumer genetic testing: two sides of the coin. PMID- 19541923 TI - p53 inhibition of AP1-dependent TFF2 expression induces apoptosis and inhibits cell migration in gastric cancer cells. AB - Overexpression of trefoil factor 2 (TFF2) is associated with increased cell migration, resistance to apoptosis, and possibly increased gastric cancer invasion. Dysregulation of p53 is frequently observed in preneoplastic conditions of the stomach. Here, we investigated the effect of p53 on the expression and function of TFF2 in gastric cancer cell lines. Gene expression was determined by reverse transcription-polymerase chain reaction, and promoter activity was assessed by dual luciferase reporter assays. Apoptosis was detected by flow cytometry, and cell migration was evaluated by the Boyden chamber assay. Exogenous expression of p53 dose dependently inhibited endogenous TFF2 mRNA, protein, and promoter activity and resulted in induction of cell apoptosis and inhibition of cell migration. Downregulation of TFF2 by small interfering RNA sensitized gastric cancer cells to drug-induced p53-dependent apoptosis. Addition of human TFF2 peptide reversed p53-dependent apoptosis and inhibition of cell migration. The p53-responsive element was mapped to an AP-1-like cis-element at 182 bp upstream of the TFF2 transcription start site. Mutation of this AP-1-like element abrogated p53-mediated inhibition of TFF2 promoter activity. Gel shift and chromatin immunoprecipitation assays demonstrated that c-Jun and c-Fos bind to this AP-1-like element. Ectopic expression of c-Jun/c-Fos or p300 or treatment of cells with phorbol 12-myristate 13-acetate (PMA) stimulated endogenous TFF2 mRNA expression and promoter activity, and p53 inhibited the effects of AP-1 and PMA on TFF2. p53 induces cell apoptosis and inhibits cell migration in part by downregulating TFF2 expression through an AP-1-like site, suggesting that TFF2 may be an important downstream target of p53. PMID- 19541921 TI - CpG methylation analysis--current status of clinical assays and potential applications in molecular diagnostics: a report of the Association for Molecular Pathology. AB - Methylation of CpG islands in gene promoter regions is a major molecular mechanism of gene silencing and underlies both cancer development and progression. In molecular oncology, testing for the CpG methylation of tissue DNA has emerged as a clinically useful tool for tumor detection, outcome prediction, and treatment selection, as well as for assessing the efficacy of treatment with the use of demethylating agents and monitoring for tumor recurrence. In addition, because CpG methylation occurs early in pre-neoplastic tissues, methylation tests may be useful as markers of cancer risk in patients with either infectious or inflammatory conditions. The Methylation Working Group of the Clinical Practice Committee of the Association of Molecular Pathology has reviewed the current state of clinical testing in this area. We report here our summary of both the advantages and disadvantages of various methods, as well as the needs for standardization and reporting. We then conclude by summarizing the most promising areas for future clinical testing in cancer molecular diagnostics. PMID- 19541924 TI - Trans fat feeding results in higher serum alanine aminotransferase and increased insulin resistance compared with a standard murine high-fat diet. AB - Diets high in trans fats are associated with an increased risk of cardiovascular disease and components of the metabolic syndrome. The influence of these toxic fatty acids on the development of nonalcoholic fatty liver disease has not been significantly examined. Therefore, we sought to compare the effect of a murine diet high in trans fat to a standard high-fat diet that is devoid of trans fats but high in saturated fats. Male AKR/J mice were fed a calorically identical trans fat diet or standard high-fat diet for 10 days, 4 wk, and 8 wk. Serum alanine aminotransferase (ALT), lipid, insulin, and leptin levels were determined and the quantitative insulin-sensitivity check index (QUICKI) was calculated as a measure of insulin resistance. Additionally, hepatic triglyceride content and gene expression of several proinflammatory genes were assessed. By 8 wk, trans fat-fed mice exhibited higher ALT values than standard high-fat-fed mice (126 +/- 16 vs. 71 +/- 7 U/l, P < 0.02) despite similar hepatic triglyceride content at each time point. Trans fat-fed mice also had increased insulin resistance compared with high-fat-fed mice at 4 and 8 wk with significantly higher insulin levels and lower QUICKI values. Additionally, hepatic interleukin-1beta (IL 1beta) gene expression was 3.6-fold higher at 4 wk (P < 0.05) and 5-fold higher at 8 wk (P < 0.05) in trans fat-fed mice compared with standard high-fat-fed mice. Trans fat feeding results in higher ALT values, increased insulin resistance, and elevated IL-1beta levels compared with standard high-fat feeding. PMID- 19541925 TI - Ca2+-independent contraction of longitudinal ileal smooth muscle is potentiated by a zipper-interacting protein kinase pseudosubstrate peptide. AB - As a regulator of smooth muscle contraction, zipper-interacting protein kinase (ZIPK) can directly phosphorylate the myosin regulatory light chains (LC20) and produce contractile force. Synthetic peptides (SM-1 and AV25) derived from the autoinhibitory region of smooth muscle myosin light chain kinase can inhibit ZIPK activity in vitro. Paradoxically, treatment of Triton-skinned ileal smooth muscle strips with AV25, but not SM-1, potentiated Ca2+-independent, microcystin- and ZIPK-induced contractions. The AV25-induced potentiation was limited to ileal and colonic smooth muscles and was not observed in rat caudal artery. Thus the potentiation of Ca2+-independent contractions by AV25 appeared to be mediated by a mechanism unique to intestinal smooth muscle. AV25 treatment elicited increased phosphorylation of LC20 (both Ser-19 and Thr-18) and myosin phosphatase-targeting subunit (MYPT1, inhibitory Thr-697 site), suggesting involvement of a Ca2+ independent LC20 kinase with coincident inhibition of myosin phosphatase. The phosphorylation of the inhibitor of myosin phosphatase, CPI-17, was not affected. The AV25-induced potentiation was abolished by pretreatment with staurosporine, a broad-specificity kinase inhibitor, but specific inhibitors of Rho-associated kinase, PKC, and MAPK pathways had no effect. When a dominant-negative ZIPK [kinase-dead ZIPK((1-320))-D161A] was added to skinned ileal smooth muscle, the potentiation of microcystin-induced contraction by AV25 was blocked. Furthermore, pretreatment of skinned ileal muscle with SM-1 abolished AV25-induced potentiation. We conclude, therefore, that, even though AV25 is an in vitro inhibitor of ZIPK, activation of the ZIPK pathway occurs following application of AV25 to permeabilized ileal smooth muscle. Finally, we propose a mechanism whereby conformational changes in the pseudosubstrate region of ZIPK permit augmentation of ZIPK activity toward LC(20) and MYPT1 in situ. AV25 or molecules based on its structure could be used in therapeutic situations to induce contractility in diseases of the gastrointestinal tract associated with hypomotility. PMID- 19541926 TI - Small intestinal efflux mediated by MRP2 and BCRP shifts sulfasalazine intestinal permeability from high to low, enabling its colonic targeting. AB - Sulfasalazine is characterized by low intestinal absorption, which essentially enables its colonic targeting and therapeutic action. The mechanisms behind this low absorption have not yet been elucidated. The purpose of this study was to investigate the role of efflux transporters in the intestinal absorption of sulfasalazine as a potential mechanism for its low small-intestinal absorption and colonic targeting following oral administration. The effects of P glycoprotein (P-gp), multidrug resistance-associated protein 2 (MRP2), and breast cancer resistance protein (BCRP) inhibitors on sulfasalazine bidirectional permeability were studied across Caco-2 cell monolayers, including dose-response analysis. Sulfasalazine in vivo permeability was then investigated in the rat jejunum by single-pass perfusion, in the presence vs. absence of inhibitors. Sulfasalazine exhibited 19-fold higher basolateral-to-apical (BL-AP) than apical to-basolateral (AP-BL) Caco-2 permeability, indicative of net mucosal secretion. MRP2 inhibitors (MK-571 and indomethacin) and BCRP inhibitors [fumitremorgin C (FTC) and pantoprazole] significantly increased AP-BL and decreased BL-AP sulfasalazine Caco-2 transport in a concentration-dependent manner. No effect was observed with the P-gp inhibitors verapamil and quinidine. The IC50 values of the specific MRP2 and BCRP inhibitors MK-571 and FTC on sulfasalazine secretion were 21.5 and 2.0 microM, respectively. Simultaneous inhibition of MRP2 and BCRP completely abolished sulfasalazine Caco-2 efflux. Without inhibitors, sulfasalazine displayed low (vs. metoprolol) in vivo intestinal permeability in the rat model. MK-571 or FTC significantly increased sulfasalazine permeability, bringing it to the low-high permeability boundary. With both MK-571 and FTC present, sulfasalazine displayed high permeability. In conclusion, efflux transport mediated by MRP2 and BCRP, but not P-gp, shifts sulfasalazine permeability from high to low, thereby enabling its colonic targeting and therapeutic action. To our knowledge, this is the first demonstration of intestinal efflux acting in favor of oral drug delivery. PMID- 19541927 TI - Amino acids in the rat intestinal lumen regulate their own absorption from a distant intestinal site. AB - Intestinal nutrient transport is altered in response to changes in dietary conditions and luminal substrate level. It is not clear, however, whether an amino acid in the intestinal lumen can acutely affect its own absorption from a distant site. Our aim is to study the effect of an amino acid present in rat small intestinal segment on its own absorption from a proximal or distal site and elucidate the underlying mechanisms. The effect of instillation of alanine (Ala) in either jejunum or ileum on its own absorption at ileal or jejunal level was examined in vivo. The modulation of this intestinal regulatory loop by the following interventions was studied: tetrodotoxin (TTX) added to Ala, subdiaphragmatic vagotomy, chemical ablation of capsaicin-sensitive primary afferent (CSPA) fibers, and IV administration of calcitonin gene-related peptide (CGRP) antagonist. In addition, the kinetics of jejunal Ala absorption and the importance of Na+-dependent transport were studied in vitro after instilling Ala in the ileum. Basal jejunal Ala absorption [0.198 +/- 0.018 micromol x cm(-1) x 20 min(-1) (means +/- SD)] was significantly decreased with the instillation of 20 mM Ala in the ileum or in an adjacent distal jejunal segment (0.12 +/- 0.015; P < 0.0001 and 0.138 +/- 0.014; P < 0.002, respectively). Comparable inhibition was observed in the presence of proline in the ileum. Moreover, basal Ala absorption from the ileum (0.169 +/- 0.025) was significantly decreased by the presence of 20 mM Ala in the jejunum (0.103 +/- 0.027; P < 0.01). The inhibitory effect on jejunal Ala absorption was abolished by TTX, subdiaphragmatic vagotomy, neonatal capsaicin treatment, and CGRP antagonism. In vitro studies showed that Ala in the ileum affects Na+-mediated transport and increases K(m) without affecting Vmax. Intraluminal amino acids control their own absorption from a distant part of the intestine, by affecting the affinity of the Na+-mediated Ala transporter, through a neuronal mechanism that involves CSPA and CGRP. PMID- 19541928 TI - High fat diet induced hepatic steatosis establishes a permissive microenvironment for colorectal metastases and promotes primary dysplasia in a murine model. AB - Non-alcoholic fatty liver disease (NAFLD), which includes steatosis and its progression to non-alcoholic steatohepatitis, is a liver disorder of increasing clinical significance. Here we characterize a murine model of high fat diet induced NAFLD with progression from liver steatosis to histological features compatible with steatohepatitis and more advanced stages of NAFLD in humans, including chronic portal inflammation, pericellular and bridging fibrosis, Mallory body formation, and bile ductular reaction. Chronic changes induced by the prolonged consumption of a high-fat diet alone culminate in the development of primary liver dysplasias. Importantly, we extend these studies to demonstrate that even the early stages of uncomplicated steatosis provide a permissive microenvironment for the growth of colon cancer cells that are metastatic to the liver. High fat diet-induced steatosis, coupled with a splenic injection model of experimental liver metastasis using syngeneic MC38 colon cancer cells, resulted in an increased number of secondary tumor nodules and metastatic burden in steatotic livers. Metastatic nodules were associated with focal peritumoral areas of infiltrating inflammatory cells and associated apoptotic cell populations. These results suggest that the modulation of specific host factors in the steatotic liver contributes to tumor progression in the microenvironment of NAFLD. PMID- 19541929 TI - Modulation of the E2F1-driven cancer cell fate by the DNA damage response machinery and potential novel E2F1 targets in osteosarcomas. AB - Osteosarcoma is the most common primary bone cancer. Mutations of the RB gene represent the most frequent molecular defect in this malignancy. A major consequence of this alteration is that the activity of the key cell cycle regulator E2F1 is unleashed from the inhibitory effects of pRb. Studies in animal models and in human cancers have shown that deregulated E2F1 overexpression possesses either "oncogenic" or "oncosuppressor" properties, depending on the cellular context. To address this issue in osteosarcomas, we examined the status of E2F1 relative to cell proliferation and apoptosis in a clinical setting of human primary osteosarcomas and in E2F1-inducible osteosarcoma cell line models that are wild-type and deficient for p53. Collectively, our data demonstrated that high E2F1 levels exerted a growth-suppressing effect that relied on the integrity of the DNA damage response network. Surprisingly, induction of p73, an established E2F1 target, was also DNA damage response-dependent. Furthermore, a global proteome analysis associated with bioinformatics revealed novel E2F1 regulated genes and potential E2F1-driven signaling networks that could provide useful targets in challenging this aggressive neoplasm by innovative therapies. PMID- 19541930 TI - Endothelin-converting enzyme-2 is increased in Alzheimer's disease and up regulated by Abeta. AB - Alzheimer's disease (AD) is thought to be caused by the accumulation of amyloid beta (Abeta) peptide within the brain. Endothelin-converting enzyme-2 (ECE-2), which is expressed in neural tissues, cleaves 'big endothelin' to produce the vasoconstrictor endothelin-1. ECE-2 also degrades Abeta. We have examined ECE-2 expression in the temporal cortex of brain tissue from patients with AD, vascular dementia, and controls. Immunohistochemistry with specific antibodies showed ECE 2 to be abundant within pyramidal neurons in both the hippocampus and neocortex, but also to be present in certain astrocytes and microglia, particularly in AD brains. Quantitative real-time PCR showed ECE-2 mRNA to be markedly elevated in AD but not in vascular dementia. ECE-2 protein concentration, measured by sandwich enzyme-linked immunosorbent assay, was also significantly elevated in AD but not in vascular dementia. Exposure of SH-SY5Y human neuroblastoma cells to monomeric or oligomeric Abeta(1-42) caused an initial decrease in ECE-2 mRNA at 4 hours, but a marked increase by 24 hours. Our findings indicate that Abeta accumulation in AD is unlikely to be caused by ECE-2 deficiency. However, ECE-2 expression is up-regulated, perhaps to minimize Abeta accumulation, but this may also be a mechanism through which endothelin-1 production is increased and cerebral blood flow is reduced in AD. Our findings suggest that endothelin-1 receptor antagonists, already licensed for treating other diseases, could be of benefit in AD therapies. PMID- 19541931 TI - Expression and functional regulation of myoglobin in epithelial cancers. AB - Myoglobin is a multifunctional heme protein that is thought to be expressed exclusively in myocytes. Its importance in both oxygen transport and free radical scavenging has been extensively characterized. We hypothesized that solid tumors could take advantage of proteins such as myoglobin to cope with hypoxic conditions and to control the metabolism of reactive oxygen and nitrogen species. We therefore sought to establish whether myoglobin might be expressed and functionally regulated in epithelial tumors that are known to face hypoxia and oxidative stress during disease progression. We analyzed the expression of myoglobin in human epithelial cancers at both transcriptional and protein levels; moreover, we investigated the expression levels of myoglobin in cancer cell lines subjected to different conditions, including hypoxia, oxidative stress, and mitogenic stimuli. We provide evidence that human epithelial tumors, including breast, lung, ovary, and colon carcinomas, express high levels of myoglobin from the earliest stages of disease development. In human cancer cells, myoglobin is induced by a variety of signals associated with tumor progression, including mitogenic stimuli, oxidative stress, and hypoxia. This study provides evidence that myoglobin, previously thought to be restricted to myocytes, is expressed at high levels by human carcinoma cells. We suggest that myoglobin expression is part of a cellular program aimed at coping with changed metabolic and environmental conditions associated with neoplastic growth. PMID- 19541932 TI - Decrease in tumor necrosis factor-alpha receptor-associated death domain results from ubiquitin-dependent degradation in obstructive renal injury in rats. AB - Increased expression levels of tumor necrosis factor-alpha (TNFalpha) is involved in tubulointerstitial cell proliferation and apoptosis in obstructive renal injury. Two TNFalpha receptors (TNFRs), TNFR1 and TNFR2, are known to exist. On TNFalpha binding, TNFR1 recruits TNFR-associated death domain (TRADD), an assembly platform to mediate TNFR1 signaling. We investigated postreceptor TRADD regulation in rat kidneys with unilateral ureteral obstruction (UUO). Whereas UUO was associated with increased expression levels of TNFalpha, TNFR1, TNFR2, and TRADD mRNAs, it resulted in the marked decrease of TRADD protein levels (which appeared at day 1 and persisted thereafter) and a slight decrease in TNFR1 protein levels at days 7 and 14. Both ubiquitination and degradation of TRADD were increased in UUO kidneys, degradation of TRADD was stimulated by TNFalpha in HK-2 cells, and TRADD degradation was suppressed by proteasome inhibitor. Inhibition of TNFalpha by soluble TNFR2, etanercept, reduced significantly, although transiently, tubular and interstitial cell proliferation, fibronectin expression, and apoptosis in UUO kidneys, and also suppressed TRADD degradation. These data suggest that the decrease in TRADD resulting from enhanced ubiquitin dependent degradation is involved in obstructive renal injury. Since TRADD is not incorporated into TNFR2-mediated TNFalpha signaling, the persistent decrease in TRADD, associated with a mild decrease in TNFR1 levels, may function, at least in part, to divert TNFalpha signals toward a TNFR2-mediated pathway in UUO kidneys. PMID- 19541933 TI - Complex integration of matrix, oxidative stress, and apoptosis in genetic emphysema. AB - Alveolar enlargement, which is characteristic of bronchopulmonary dysplasia, congenital matrix disorders, and cigarette smoke-induced emphysema, is thought to result from enhanced inflammation and ensuing excessive matrix proteolysis. Although there is recent evidence that cell death and oxidative stress punctuate these diseases, the mechanistic link between abnormal lung extracellular matrix and alveolar enlargement is lacking. We hypothesized that the tight-skin (TSK) mouse, which harbors a spontaneous internal duplication in the microfibrillar glycoprotein fibrillin-1, might show whether matrix alterations are sufficient to promote oxidative stress and cell death, injury cascades central to the development of clinical emphysema. We observed no evidence of increased metalloprotease activation by histochemical and zymographic methods. We did find initial oxidative stress followed by increased apoptosis in the postnatal TSK lung. Both blunted antioxidant production and reduced extracellular superoxide dismutase activity were evident in the neonatal lung. High-dose antioxidant treatment with N-acetylcysteine improved airspace caliber and attenuated oxidative stress and apoptosis in neonatal and adult TSK mice. These data establish that an abnormal extracellular matrix without overt elastolysis is sufficient to confer susceptibility to postnatal normoxia, reminiscent of bronchopulmonary dysplasia. The resultant oxidative stress and apoptosis culminate in profound airspace enlargement. The TSK lung exemplifies the critical interplay between extracellular matrix, oxidative stress, and cell-death cascades that may contribute to genetic and acquired airspace enlargement. PMID- 19541934 TI - Complement factor h is critical in the maintenance of retinal perfusion. AB - Vascular pathologies are known to be associated with age-related macular degeneration. Recently, age-related macular degeneration was associated with a single-nucleotide substitution of the complement factor H (CFH) gene, part of the alternative pathway of the complement system, a critical element in the innate immune response. Such polymorphisms are found in more than 50% of cases of age related macular degeneration. Here we show that the absence of CFH causes an autoimmune response that targets the vascular endothelium of both the inner and outer retinal vascular networks. In CFH-knockout (cfh(-/-)) mice, C3 and C3b, key components of the complement system, are progressively deposited on retinal vessels, which subsequently become restricted and wither, resulting in a reduction of retinal blood supply. This result leads to increased oxygen stress. While such effects are not systemic, these structural changes are mirrored in functional changes with a substantial decline in retinal blood flow dynamics. When the system is challenged functionally by laser-induced choroidal neovascularization, fluorescein leakage was significantly smaller in cfh(-/-) mice compared with controls, likely due to reduced retinal perfusion. These data reveal that in both the presence and absence of exogenous challenge to the innate immune system, CFH is required to maintain normal levels of retinal perfusion. It is likely that C3 and C3b accumulation in the aged CFH-deficient retina is associated with complement-mediated retinal endothelium destruction. PMID- 19541935 TI - Up-regulation of soluble Axl and Mer receptor tyrosine kinases negatively correlates with Gas6 in established multiple sclerosis lesions. AB - Multiple sclerosis is a disease that is characterized by inflammation, demyelination, and axonal damage; it ultimately forms gliotic scars and lesions that severely compromise the function of the central nervous system. Evidence has shown previously that altered growth factor receptor signaling contributes to lesion formation, impedes recovery, and plays a role in disease progression. Growth arrest-specific protein 6 (Gas6), the ligand for the TAM receptor tyrosine kinase family, consisting of Tyro3, Axl, and Mer, is important for cell growth, survival, and clearance of debris. In this study, we show that levels of membrane bound Mer (205 kd), soluble Mer ( approximately 150 kd), and soluble Axl (80 kd) were all significantly elevated in homogenates from established multiple sclerosis lesions comprised of both chronic active and chronic silent lesions. Whereas in normal tissue Gas6 positively correlated with soluble Axl and Mer, there was a negative correlation between Gas6 and soluble Axl and Mer in established multiple sclerosis lesions. In addition, increased levels of soluble Axl and Mer were associated with increased levels of mature ADAM17, mature ADAM10, and Furin, proteins that are associated with Axl and Mer solubilization. Soluble Axl and Mer are both known to act as decoy receptors and block Gas6 binding to membrane-bound receptors. These data suggest that in multiple sclerosis lesions, dysregulation of protective Gas6 receptor signaling may prolong lesion activity. PMID- 19541938 TI - Osteoid osteoma of the odontoid process of the axis associated with atlanto-axial fusion. AB - Osteoid osteoma is a primary benign bone tumour with characteristic imaging and pathological features. In about 10% of the cases it involves the spine, especially the posterior elements. We report a pathologically proven case of osteoid osteoma of the odontoid process of the second cervical vertebra associated with atlanto-axial fusion. The case was diagnosed pre-operatively using plain radiography, multiplanar helical CT, contrast-enhanced MRI and radioisotope bone scanning. PMID- 19541936 TI - NOLC1, an enhancer of nasopharyngeal carcinoma progression, is essential for TP53 to regulate MDM2 expression. AB - Nasopharyngeal carcinoma (NPC) is one of the most common cancers among Chinese living in South China, Singapore, and Taiwan. At present, its etiological factors are not well defined. To identify which genetic alterations might be involved in NPC pathogenesis, we identified genes that were differentially expressed in NPC cell lines and normal nasomucosal cells using subtractive hybridization and microarray analysis. Most NPC cell lines and biopsy specimens were found to have higher expression levels of the gene encoding nucleolar and coiled-body phosphoprotein 1 (NOLC1) as compared with normal cells. Severe combined immunodeficiency mice bearing NPC xenografts derived from NOLC1-short hairpin-RNA transfected animals were found to have 82% lower levels of tumor growth than control mice as well as marked tumor cell apoptosis. Measuring the expression levels of genes related to cell growth, apoptosis, and angiogenesis, we found that the MDM2 gene was down-regulated in the transfectants. Both co-transfection and chromatin immunoprecipitation experiments showed that tumor protein 53 regulated expression of the MDM2 gene requires co-activation of NOLC1. These findings suggest that NOLC1 plays a role in the regulation of tumorigenesis of NPC and demonstrate that both NOLC1 and tumor protein 53 work together synergistically to activate the MDM2 promoter in NPC cells. PMID- 19541937 TI - Modulation of TGFbeta1-dependent myofibroblast differentiation by hyaluronan. AB - Myofibroblasts are contractile cells that are characterized by the expression of alpha-smooth muscle actin and mediate the closure of wounds and the formation of collagen-rich scars. Their presence in organs such as lungs, liver, and kidney has long been established as a marker of progressive fibrosis. The transforming growth factor beta(1)-driven differentiation of fibroblasts is a major source of myofibroblasts, and recent data have shown that hyaluronan is a major modulator of this process. This study examines this differentiation mechanism in more detail. Transforming growth factor beta(1)-dependent differentiation to the myofibroblastic phenotype was antagonized by the inhibition of hyaluronan synthesis, confirming that hyaluronan was necessary for differentiation. This response, however, was not reproduced by simply adding hyaluronan to fibroblasts, as the results implicated hyaladherins, as well as the macromolecular assembly of de novo hyaluronan, as essential in this process. We previously suggested that there is a relocalization of lipid-raft components during myofibroblastic differentiation. The present study demonstrates that the hyaluronan receptor CD44, the hyaluronidase HYAL 2, and the transforming growth factor beta(1) receptor ALK5 all relocalized from raft to non-raft locations, which was reversed by the addition of exogenous hyaluronan. These data highlight a role for endogenous hyaluronan in the mediation of myofibroblastic differentiation. While hyaluronan synthesis was both essential and necessary for differentiation, exogenously provided hyaluronan antagonized differentiation, underscoring a pathological role for hyaluronan in such cell fate processes. PMID- 19541939 TI - Cystic meningioangiomatosis in neurofibromatosis type 2: an MRI-pathological study. AB - We report cerebral cystic meningioangiomatosis in a patient with neurofibromatosis type 2. An 18-year-old woman presented with progressive hemiparesis secondary to a meningioma at the foramen magnum. Her MR examination also demonstrated three small cortical and subcortical cystic lesions. She underwent surgery for the meningioma, but died from brainstem infarction. Post mortem histopathological examination of the cystic lesions showed enlarged subcortical perivascular spaces with overlying meningioangiomatosis. The unusual features and possible pathogenesis are discussed. PMID- 19541940 TI - Multiple gastrointestinal stromal tumours presenting as a haemoperitoneum in a patient with Type 1 neurofibromatosis: MDCT findings. AB - We report multidetector row CT images of multiple gastrointestinal tumours in a patient with Type 1 neurofibromatosis who presented with haemoperitoneum. The use of CT demonstrated multiple, variably sized, ovoid, relatively well-enhancing masses in the intramural or subserosal location of the jejunum. The largest mass that showed a haemorrhagic necrosis caused haemoperitoneum. These imaging features are unique and differ from previous imaging features that have been reported in the radiological literature. PMID- 19541941 TI - Postural change in ventilation and perfusion secondary to a thoracic scoliosis with complete resolution after surgery. AB - We present the case of a child with a thoracic scoliosis causing respiratory impairment in whom pre-surgical ventilation-perfusion lung scintigraphy in different postures was able to predict improvement in ventilation and perfusion after surgery. PMID- 19541942 TI - Left-sided gallbladder: a complicated percutaneous cholecystostomy and subsequent hepatic embolisation. AB - A 68-year-old male patient with chronic hypertension, diabetes mellitus and chronic renal failure was diagnosed with acute calculous cholecystitis. A percutaneous cholecystostomy using a transperitoneal approach was performed after two failed attempts with a right-sided transhepatic approach. Subsequent hepatic embolisation was performed for the treatment of haemoperitoneum due to hepatic injury after the percutaneous cholecystostomy. The presence of a left-sided gallbladder was confirmed by laparoscopic cholecystectomy after 1 week. Prior identification of this anomaly would have prevented hepatic injury through the use of a cautious procedure against mobility or careful selection of the approach routes. In conclusion, the transperitoneal approach can be easier or more preferable to perform for a percutaneous cholecystostomy of a left-sided gallbladder. PMID- 19541943 TI - Ultrasound and CT findings in hepatic and pancreatic parenchyma in acute schistosomiasis. AB - A case of acute toxaemic schistosomiasis is presented. Abdominal ultrasound examination revealed hepatosplenomegaly, lymphadenomegaly, heterogeneous hepatic parenchyma and heterogeneous focal lesions in the pancreas. CT confirmed lymph node enlargement and hepatosplenomegaly, and showed multiple small focal nodular lesions in the liver and focal lesions in the pancreas. To our knowledge, this is the first description of pancreatic lesions associated with acute Schistosoma mansoni infection. PMID- 19541944 TI - CT lung cancer screening in the UK. AB - Lung cancer is the most common cause of cancer-related death in the UK. Despite aggressive primary prevention measures and improved medical care, the 5-year survival rate is less than 10% for patients in the UK who present with symptoms. The possibility of CT screening for lung cancer provides some hope of reducing mortality. However, the case for screening remains unproven. This article explores the issues surrounding lung cancer screening in the context of historical studies, trials in progress and tentative plans for a UK CT lung cancer screening trial. PMID- 19541945 TI - Differential diagnosis of hyperintense cerebrospinal fluid on fluid-attenuated inversion recovery images of the brain. Part II: non-pathological conditions. AB - The fluid-attenuated inversion recovery imaging sequence is a widely used MRI sequence of the brain. It is an inversion recovery pulse sequence, designed to suppress signals from the cerebrospinal fluid. It is highly sensitive for detection of lesions adjacent to or within the cerebrospinal fluid, associated with T(2) prolongation or T(1) shortening. The term "hyperintense cerebrospinal fluid" is used to describe failed suppression or hyperintensity of cerebrospinal fluid on fluid-attenuated inversion recovery imaging of the brain. It is often encountered in many important pathological conditions, including subarachnoid haemorrhage, meningitis and leptomeningeal metastasis. However, certain non pathological states in which there is no definite cerebrospinal fluid abnormality can also present with hyperintense cerebrospinal fluid. Correct interpretation of abnormalities is important to arrive at an appropriate diagnosis. This pictorial review provides fluid-attenuated inversion recovery images of hyperintense cerebrospinal fluid of the brain and describes distinguishing features, focusing on non-pathological conditions. PMID- 19541946 TI - Letter to the editor concerning "320-slice CT neuroimaging: initial clinical experience and image quality evaluation" (Siebert E et al: Br J Radiol 2009;82:561-70). PMID- 19541947 TI - A longitudinal study of policy effect (smoke-free legislation) on smoking norms: ITC Scotland/United Kingdom. AB - INTRODUCTION: The longitudinal ITC Scotland/U.K. survey was used to investigate adult smokers' support for smoke-free legislation and whether this support was associated with higher quit intentions at follow-up, either directly or indirectly, via the mediation of perceived social unacceptability of smoking. METHODS: Structural equation modeling was employed to compare differences between the two samples (507 adult smokers from Scotland and 507 from the rest of the United Kingdom) across two waves (February/March 2006 and March 2007). During these two waves, a smoking ban was introduced in Scotland but not the rest of the United Kingdom. RESULTS: For smokers in both samples, support for smoke-free legislation at baseline significantly heightened perceived unacceptability of smoking, although perceptions of unacceptability were somewhat stronger in Scotland than the rest of the United Kingdom postban. Unlike the rest of the United Kingdom, support for a ban at baseline among smokers in Scotland was associated with higher quit intentions at follow-up. For both samples, quit intentions were significantly associated with heightened perceived unacceptability at follow-up. The overall variance explained in quit intentions was greater in Scotland than in the rest of the United Kingdom but not significantly so. DISCUSSION: Support for smoke-free legislation at baseline significantly increased support at follow-up for both samples. However, this did not independently increase quit intentions among smokers from both Scotland and the rest of the United Kingdom. The findings suggest that normative influences are one of the mechanisms through which comprehensive smoke-free legislation influences quit intentions. PMID- 19541948 TI - Feasibility of an exercise counseling intervention for depressed women smokers. AB - INTRODUCTION: Depressive symptoms negatively impact smoking abstinence. However, few interventions have been targeted to smokers with current depression. Exercise improves mood and may benefit depressed smokers. This pilot study investigated the feasibility of an exercise intervention for depressed female smokers (Center for Epidemiological Studies Depression Scale [CES-D] score > or =16). METHODS: Participants (M = 41 years, 98% White) were randomized to 10 weeks of individually delivered exercise counseling (n = 30) or a health education contact control condition (n = 30). All participants received nicotine patch therapy and behavioral counseling for smoking cessation. RESULTS: The intervention was feasible as indicated by ability to recruit participants, exercise counseling session attendance (M = 7.6 of 10 sessions attended), and significant increase in exercise frequency and stage of change from baseline to end of treatment (EOT) (Week 10). Participant attrition rate was 35% by Week 10 but did not differ significantly between groups. Smoking abstinence rates at Week 10, using intention-to-treat analysis, were 17% for exercise counseling participants and 23% for health education participants (p = .75). DISCUSSION: An exercise counseling intervention was found to be feasible for depressed women smokers. More intensive intervention may be needed to increase smoking abstinence rates, and methods should be refined to reduce participant burden and attrition. PMID- 19541949 TI - Consumer awareness and attitudes related to new potential reduced-exposure tobacco product brands. AB - INTRODUCTION: In recent years, there has been a proliferation of potential reduced-exposure tobacco products (PREPs) marketed that claim to be less harmful or less addictive, compared with conventional cigarettes. Tobacco control scientists have raised concerns about the potential adverse impact of marketing of these products for smoking prevention and cessation efforts. Although these products have not been widely used among smokers, there are few data available on consumers' awareness and attitudes toward these products. METHODS: Data were obtained from the 2003 and 2005 Health Information National Trends Survey, a nationally representative telephone survey of adults 18 years and older regarding health communication and associated beliefs and behaviors. Our study population consisted of 6,369 respondents in 2003 and 5,586 respondents in 2005, of whom 19% were current smokers and 28% were former smokers. RESULTS: In 2005, 45% of respondents had heard of at least one PREP product, while only 4.8% had actually tried one. Awareness and use were substantially higher among current smokers (55.6% and 12.7%). Awareness was highest for Marlboro Ultra Smooth (MUS) (30.2%), Eclipse (18.2%), Quest (7.8%), and Ariva (5.4%), while less than 2% for any other product. Of respondents who had tried a PREP, 50% cited harm reduction or assistance in quitting as a reason for trying the product and 30% believed that the product was less harmful than their usual brand. In the combined 2003 and 2005 dataset, 54.4% of current smokers stated that they would be "very" or "somewhat" interested in trying a cigarette advertised as less harmful, while only 3.2% of former smokers and 1.1% of never-smokers were interested. Among current smokers, interest was higher in females and non-Hispanic Whites, and among daily smokers, those who smoked 20 or more cigarettes per day and those who were not considering quitting. Smokers interested in PREPs were substantially more likely to rate their perceived lung cancer risk as high (40.3% vs. 8.3%) and to worry frequently about developing lung cancer (19.7% vs. 4%). DISCUSSION: These results suggest that there is a substantial level of interest among current smokers in cigarettes marketed with claims of reduced exposure or harm. Of particular concern is that "health conscious" smokers and heavy smokers not planning to quit may be especially vulnerable to PREP marketing messages and view such products as an alternative to smoking cessation. PMID- 19541950 TI - Smokers of illicit tobacco report significantly worse health than other smokers. AB - INTRODUCTION: The aim of this study was to ascertain whether the health of past and current smokers of illicit tobacco (chop-chop) differs from that of smokers of licit tobacco. METHODS: The design was a telephone survey, stratified by state, using computer aided telephone interviewing, with households selected by random digit dialing from the telephone white pages. Setting was all Australian states and territories, 1,621 regular tobacco smokers aged 18+ years. Measures were social and personal characteristics of tobacco smokers, smoking histories and patterns, and health status (SF-8 and disability weights). Binary logistic regression was used to identify variables associated with current and lifetime chop-chop use. RESULTS: Compared with licit-only tobacco smokers, current users of chop-chop had significantly greater odds of beginning smoking aged <16 years (odds ratio [OR] 1.65, 95% CI = 1.09-2.50), of reporting below-average social functioning (OR 1.61, 95% CI = 1.06-2.44), and of a measurable disability (OR 1.95, 95% CI = 1.08-3.51). Lifetime chop-chop users were relatively likely to be less than 45 years of age (OR 1.82, 95% CI = 1.38-2.39), report below-average mental health (OR 1.61, 95% CI = 1.22-2.13) and above-average bodily pain (OR 1.40, 95% CI = 1.06-1.85), smoke more than 120 cigarettes/week (OR 1.39, 95% CI = 1.06-1.83), and to have begun smoking aged <16 years (OR 1.33, 95% CI = 1.01 1.75). DISCUSSION: Current and lifetime users of chop-chop report significantly worse health than smokers of licit tobacco. Investigation of how to communicate this finding to current and potential chop-chop smokers is warranted. PMID- 19541951 TI - Estimating smokers' mouth-level exposure to select mainstream smoke constituents from discarded cigarette filter butts. AB - INTRODUCTION: Standardized machine smoking measurements are poor predictors of exposure. We have refined a method using the solanesol deposited in discarded cigarette butts as a marker for estimating deliveries of mainstream smoke constituents. Developing a fast and accurate method for measuring solanesol in cigarette filters to assess tobacco smoke intake could provide a way to assess how people smoke under natural conditions. We have developed and validated a new, lower-cost, high-throughput method to measure the solanesol content in discarded cigarette filter butts and correlated these measurements with mainstream smoke deliveries of nicotine and tobacco-specific nitrosamines (TSNAs). METHODS: Cigarettes were machine smoked under a variety of conditions to cover a wide range of nicotine deliveries and solanesol levels in the spent cigarette filter. Following machine smoking, a 1-cm portion of filter material, measured from the mouth end, was removed from the cigarette butts for analysis. Although an isotopically labeled solanesol analog is currently not commercially available, we achieved excellent quantitative results using a structurally similar compound, geranylgeraniol, as an internal standard (IS). After spiking with IS and solvent extracted, solanesol extracts were then analyzed using liquid chromatography coupled with a single-quadrupole mass analyzer. Analysis was carried out using manual preparation as well as a high-throughput 48-well format using automated liquid handlers. RESULTS: Recoveries of solanesol from cigarette butts exceeded 95% with excellent precision and exhibited excellent linearity for both preparation methods. In addition, we show that the mouth-level exposure for both nicotine and TSNAs may be estimated by their relation to the solanesol retained in the cigarette filter. DISCUSSION: We believe that this method provides excellent versatility and throughput for the estimation of mouth-level exposure to a wide range of toxins in cigarette smoke under naturalistic conditions. In addition, this method allows a far more accurate measure of exposure both from a single cigarette as well as from daily smoking. PMID- 19541952 TI - Consistency between satellite-derived and modeled estimates of the direct aerosol effect. AB - In the Intergovernmental Panel on Climate Change Fourth Assessment Report, the direct aerosol effect is reported to have a radiative forcing estimate of -0.5 Watt per square meter (W m(-2)), offsetting the warming from CO2 by almost one third. The uncertainty, however, ranges from -0.9 to -0.1 W m(-2), which is largely due to differences between estimates from global aerosol models and observation-based estimates, with the latter tending to have stronger (more negative) radiative forcing. This study demonstrates consistency between a global aerosol model and adjustment to an observation-based method, producing a global and annual mean radiative forcing that is weaker than -0.5 W m(-2), with a best estimate of -0.3 W m(-2). The physical explanation for the earlier discrepancy is that the relative increase in anthropogenic black carbon (absorbing aerosols) is much larger than the overall increase in the anthropogenic abundance of aerosols. PMID- 19541953 TI - Successful conservation of a threatened Maculinea butterfly. AB - Globally threatened butterflies have prompted research-based approaches to insect conservation. Here, we describe the reversal of the decline of Maculinea arion (Large Blue), a charismatic specialist whose larvae parasitize Myrmica ant societies. M. arion larvae were more specialized than had previously been recognized, being adapted to a single host-ant species that inhabits a narrow niche in grassland. Inconspicuous changes in grazing and vegetation structure caused host ants to be replaced by similar but unsuitable congeners, explaining the extinction of European Maculinea populations. Once this problem was identified, UK ecosystems were perturbed appropriately, validating models predicting the recovery and subsequent dynamics of the butterfly and ants at 78 sites. The successful identification and reversal of the problem provides a paradigm for other insect conservation projects. PMID- 19541954 TI - Translocator protein (18 kD) as target for anxiolytics without benzodiazepine like side effects. AB - Most antianxiety drugs (anxiolytics) work by modulating neurotransmitters in the brain. Benzodiazepines are fast and effective anxiolytic drugs; however, their long-term use is limited by the development of tolerance and withdrawal symptoms. Ligands of the translocator protein [18 kilodaltons (kD)] may promote the synthesis of endogenous neurosteroids, which also exert anxiolytic effects in animal models. Here, we found that the translocator protein (18 kD) ligand XBD173 enhanced gamma-aminobutyric acid-mediated neurotransmission and counteracted induced panic attacks in rodents in the absence of sedation and tolerance development. XBD173 also exerted antipanic activity in humans and, in contrast to benzodiazepines, did not cause sedation or withdrawal symptoms. Thus, translocator protein (18 kD) ligands are promising candidates for fast-acting anxiolytic drugs with less severe side effects than benzodiazepines. PMID- 19541955 TI - Penumbral structure and outflows in simulated sunspots. AB - Sunspots are concentrations of magnetic field on the visible solar surface that strongly affect the convective energy transport in their interior and surroundings. The filamentary outer regions (penumbrae) of sunspots show systematic radial outward flows along channels of nearly horizontal magnetic field. These flows were discovered 100 years ago and are present in all fully developed sunspots. By using a comprehensive numerical simulation of a sunspot pair, we show that penumbral structures with such outflows form when the average magnetic field inclination to the vertical exceeds about 45 degrees. The systematic outflows are a component of the convective flows that provide the upward energy transport and result from anisotropy introduced by the presence of the inclined magnetic field. PMID- 19541957 TI - Science journalism goes global. PMID- 19541956 TI - Functional amyloids as natural storage of peptide hormones in pituitary secretory granules. AB - Amyloids are highly organized cross-beta-sheet-rich protein or peptide aggregates that are associated with pathological conditions including Alzheimer's disease and type II diabetes. However, amyloids may also have a normal biological function, as demonstrated by fungal prions, which are involved in prion replication, and the amyloid protein Pmel17, which is involved in mammalian skin pigmentation. We found that peptide and protein hormones in secretory granules of the endocrine system are stored in an amyloid-like cross-beta-sheet-rich conformation. Thus, functional amyloids in the pituitary and other organs can contribute to normal cell and tissue physiology. PMID- 19541958 TI - Swine flu. After delays, WHO agrees: the 2009 pandemic has begun. PMID- 19541959 TI - Public health. Expanded U.S. drug agency to control tobacco. PMID- 19541960 TI - U.S. STEM education. Report calls for grassroots but comprehensive changes. PMID- 19541963 TI - China. After outcry, government backpedals over internet-filtering software. PMID- 19541962 TI - Arms control. Verification experts puzzled over North Korea's nuclear test. PMID- 19541964 TI - Law of the sea. A final push to divvy up the sea by all the rules. PMID- 19541966 TI - Europe. Italy's MIT grows, and so does controversy over it. PMID- 19541967 TI - American Astronomical Society 214th meeting, 7-11 June 2009, Pasadena, California. Star studies yield better yardsticks. PMID- 19541968 TI - American Astronomical Society 214th meeting, 7-11 June 2009, Pasadena, California. Dark-matter model multiplies mass of galactic black holes. PMID- 19541969 TI - Genetics. The promise of a cure: 20 years and counting. PMID- 19541970 TI - China. Radio astronomers go for high gain with mammoth telescope. PMID- 19541971 TI - Economic recovery. ISO ... 3.5 million U.S. jobs. PMID- 19541972 TI - Physics. Is quantum mechanics tried, true, wildly successful, and wrong? PMID- 19541973 TI - Cataloguing names the old-fashioned way. PMID- 19541974 TI - Venezuelan science at risk. PMID- 19541975 TI - Machines fall short of revolutionary science. PMID- 19541976 TI - Livestock genomics in developing countries. PMID- 19541977 TI - Too much quantification hinders creativity. PMID- 19541978 TI - Comment on "Experimental test of self-shielding in vacuum ultraviolet photodissociation of CO". AB - Chakraborty et al. (Reports, 5 September 2008, p. 1328) concluded that an anomalously enriched atomic oxygen reservoir can be generated through carbon monoxide photodissociation without self-shielding. We show that this conclusion is based on the incorrect assumption that the spectral shifts of the 97.03 nanometers and 107.61-nanometers vibrational bands for C16O, C17O, and C18O are negligible and point out shortcomings of the low-resolution light source used in their experiments. PMID- 19541979 TI - Comment on "Experimental test of self-shielding in vacuum ultraviolet photodissociation of CO". AB - Chakraborty et al. (Reports, 5 September 2008, p. 1328) suggested that experimental results provide support for CO photodissociation having caused the oxygen isotope ratio associated with the early solar nebula. We point out that further analysis is required before other mechanisms, such as self-shielding, are shown to be of little importance. PMID- 19541980 TI - Comment on "Experimental test of self-shielding in vacuum ultraviolet photodissociation of CO". AB - Chakraborty et al. (Reports, 5 September 2008, p. 1328) demonstrated very large, wavelength-dependent mass-independent isotopic effects during carbon monoxide (CO) photodissociation and argued that self-shielding in CO was not responsible. We suggest that variations in band oscillator strengths and linewidths among CO isotopologs are responsible for most of the wavelength dependence observed and that the reported experiments confirm the importance of self-shielding during CO photodissociation. PMID- 19541981 TI - Agriculture. Nutrient imbalances in agricultural development. PMID- 19541982 TI - Microbiology. No place too cold. PMID- 19541983 TI - Chemistry. Squeezing the water out of HCl(aq). PMID- 19541984 TI - Ecology. Thriving in salt. PMID- 19541985 TI - Immunology. A chronic need for IL-21. PMID- 19541986 TI - Chemistry. Extracting potentials from spectra. PMID- 19541987 TI - Neuroscience. Bridging the gap and staying local. PMID- 19541988 TI - Evolution. Uniting alignments and trees. PMID- 19541989 TI - Graphene: status and prospects. AB - Graphene is a wonder material with many superlatives to its name. It is the thinnest known material in the universe and the strongest ever measured. Its charge carriers exhibit giant intrinsic mobility, have zero effective mass, and can travel for micrometers without scattering at room temperature. Graphene can sustain current densities six orders of magnitude higher than that of copper, shows record thermal conductivity and stiffness, is impermeable to gases, and reconciles such conflicting qualities as brittleness and ductility. Electron transport in graphene is described by a Dirac-like equation, which allows the investigation of relativistic quantum phenomena in a benchtop experiment. This review analyzes recent trends in graphene research and applications, and attempts to identify future directions in which the field is likely to develop. PMID- 19541990 TI - Sexual intercourse involving giant sperm in Cretaceous ostracode. AB - Reproduction with giant sperm occurs in distinct groups scattered over the animal kingdom. Although experiments in Drosophila assessed the influence of different selection pressures on this character, no information was available on its long term stability. Sub-micrometer-resolution synchrotron quantitative phase tomography (holotomography) of exceptionally well-preserved three-dimensional Cretaceous ostracode fossils from the Brazilian Santana Formation indicates that ostracode reproduction with giant sperm persisted for at least over the past 100 million years. Remnants of the male sperm pumps as well as giant, inflated female sperm receptacles evidence that, despite high costs, reproduction with giant sperm can be an evolutionary successful strategy. PMID- 19541991 TI - Solar-like oscillations in a massive star. AB - Seismology of stars provides insight into the physical mechanisms taking place in their interior, with modes of oscillation probing different layers. Low-amplitude acoustic oscillations excited by turbulent convection were detected four decades ago in the Sun and more recently in low-mass main-sequence stars. Using data gathered by the Convection Rotation and Planetary Transits mission, we report here on the detection of solar-like oscillations in a massive star, V1449 Aql, which is a known large-amplitude (beta Cephei) pulsator. PMID- 19541992 TI - Colloidal quantum-dot photodetectors exploiting multiexciton generation. AB - Multiexciton generation (MEG) has been indirectly observed in colloidal quantum dots, both in solution and the solid state, but has not yet been shown to enhance photocurrent in an optoelectronic device. Here, we report a class of solution processed photoconductive detectors, sensitive in the ultraviolet, visible, and the infrared, in which the internal gain is dramatically enhanced for photon energies Ephoton greater than 2.7 times the quantum-confined bandgap Ebandgap. Three thin-film devices with different quantum-confined bandgaps (set by the size of their constituent lead sulfide nanoparticles) show enhancement determined by the bandgap-normalized photon energy, Ephoton/Ebandgap, which is a clear signature of MEG. The findings point to a valuable role for MEG in enhancing the photocurrent in a solid-state optoelectronic device. We compare the conditions on carrier excitation, recombination, and transport for photoconductive versus photovoltaic devices to benefit from MEG. PMID- 19541993 TI - Aggregation-induced dissociation of HCl(H2O)4 below 1 K: the smallest droplet of acid. AB - Acid dissociation and the subsequent solvation of the charged fragments at ultracold temperatures in nanoenvironments, as distinct from ambient bulk water, are relevant to atmospheric and interstellar chemistry but remain poorly understood. Here we report the experimental observation of a nanoscopic aqueous droplet of acid formed within a superfluid helium cluster at 0.37 kelvin. High resolution mass-selective infrared laser spectroscopy reveals that successive aggregation of the acid HCl with water molecules, HCl(H2O)n, readily results in the formation of hydronium at n = 4. Accompanying ab initio simulations show that undissociated clusters assemble by stepwise water molecule addition in electrostatic steering arrangements up to n = 3. Adding a fourth water molecule to the ringlike undissociated HCl(H2O)3 then spontaneously yields the compact dissociated H3O+(H2O)3Cl- ion pair. This aggregation mechanism bypasses deep local energy minima on the n = 4 potential energy surface and offers a general paradigm for reactivity at ultracold temperatures. PMID- 19541994 TI - Atmospheric carbon dioxide concentration across the mid-Pleistocene transition. AB - The dominant period of Pleistocene glacial cycles changed during the mid Pleistocene from 40,000 years to 100,000 years, for as yet unknown reasons. Here we present a 2.1-million-year record of sea surface partial pressure of CO2 (Pco2), based on boron isotopes in planktic foraminifer shells, which suggests that the atmospheric partial pressure of CO2 (pco2) was relatively stable before the mid-Pleistocene climate transition. Glacial Pco2 was approximately 31 microatmospheres higher before the transition (more than 1 million years ago), but interglacial Pco2 was similar to that of late Pleistocene interglacial cycles (<450,000 years ago). These estimates are consistent with a close linkage between atmospheric CO2 concentration and global climate, but the lack of a gradual decrease in interglacial Pco2 does not support the suggestion that a long-term drawdown of atmospheric CO2 was the main cause of the climate transition. PMID- 19541995 TI - Fossil plant relative abundances indicate sudden loss of Late Triassic biodiversity in East Greenland. AB - The pace of Late Triassic (LT) biodiversity loss is uncertain, yet it could help to decipher causal mechanisms of mass extinction. We investigated relative abundance distributions (RADs) of six LT plant assemblages from the Kap Stewart Group, East Greenland, to determine the pace of collapse of LT primary productivity. RADs displayed not simply decreases in the number of taxa, but decreases in the number of common taxa. Likelihood tests rejected a hypothesis of continuously declining diversity. Instead, the RAD shift occurred over the upper two-to-four fossil plant assemblages and most likely over the last three (final 13 meters), coinciding with increased atmospheric carbon dioxide concentration and global warming. Thus, although the LT event did not induce mass extinction of plant families, it accompanied major and abrupt change in their ecology and diversity. PMID- 19541996 TI - Rapid and accurate large-scale coestimation of sequence alignments and phylogenetic trees. AB - Inferring an accurate evolutionary tree of life requires high-quality alignments of molecular sequence data sets from large numbers of species. However, this task is often difficult, slow, and idiosyncratic, especially when the sequences are highly diverged or include high rates of insertions and deletions (collectively known as indels). We present SATe (simultaneous alignment and tree estimation), an automated method to quickly and accurately estimate both DNA alignments and trees with the maximum likelihood criterion. In our study, it improved tree and alignment accuracy compared to the best two-phase methods currently available for data sets of up to 1000 sequences, showing that coestimation can be both rapid and accurate in phylogenetic studies. PMID- 19541997 TI - Merkel cells are essential for light-touch responses. AB - The peripheral nervous system detects different somatosensory stimuli, including pain, temperature, and touch. Merkel cell-neurite complexes are touch receptors composed of sensory afferents and Merkel cells. The role that Merkel cells play in light-touch responses has been the center of controversy for over 100 years. We used Cre-loxP technology to conditionally delete the transcription factor Atoh1 from the body skin and foot pads of mice. Merkel cells are absent from these areas in Atoh1(CKO) animals. Ex vivo skin/nerve preparations from Atoh1(CKO) animals demonstrate complete loss of the characteristic neurophysiologic responses normally mediated by Merkel cell-neurite complexes. Merkel cells are, therefore, required for the proper encoding of Merkel receptor responses, suggesting that these cells form an indispensible part of the somatosensory system. PMID- 19541999 TI - Oceanobacter-related bacteria are important for the degradation of petroleum aliphatic hydrocarbons in the tropical marine environment. AB - Petroleum-hydrocarbon-degrading bacteria were obtained after enrichment on crude oil (as a 'chocolate mousse') in a continuous supply of Indonesian seawater amended with nitrogen, phosphorus and iron nutrients. They were related to Alcanivorax and Marinobacter strains, which are ubiquitous petroleum-hydrocarbon degrading bacteria in marine environments, and to Oceanobacter kriegii (96.4-96.5 % similarities in almost full-length 16S rRNA gene sequences). The Oceanobacter related bacteria showed high n-alkane-degrading activity, comparable to that of Alcanivorax borkumensis strain SK2. On the other hand, Alcanivorax strains exhibited high activity for branched-alkane degradation and thus could be key bacteria for branched-alkane biodegradation in tropical seas. Oceanobacter related bacteria became most dominant in microcosms that simulated a crude oil spill event with Indonesian seawater. The dominance was observed in microcosms that were unamended or amended with fertilizer, suggesting that the Oceanobacter related strains could become dominant in the natural tropical marine environment after an accidental oil spill, and would continue to dominate in the environment after biostimulation. These results suggest that Oceanobacter-related bacteria could be major degraders of petroleum n-alkanes spilt in the tropical sea. PMID- 19542000 TI - Inactivation of the ilvB1 gene in Mycobacterium tuberculosis leads to branched chain amino acid auxotrophy and attenuation of virulence in mice. AB - Acetohydroxyacid synthase (AHAS) is the first enzyme in the branched-chain amino acid biosynthesis pathway in bacteria. Bioinformatics analysis revealed that the Mycobacterium tuberculosis genome contains four genes (ilvB1, ilvB2, ilvG and ilvX) coding for the large catalytic subunit of AHAS, whereas only one gene (ilvN or ilvH) coding for the smaller regulatory subunit of this enzyme was found. In order to understand the physiological role of AHAS in survival of the organism in vitro and in vivo, we inactivated the ilvB1 gene of M. tuberculosis. The mutant strain was found to be auxotrophic for all of the three branched-chain amino acids (isoleucine, leucine and valine), when grown with either C(6) or C(2) carbon sources, suggesting that the ilvB1 gene product is the major AHAS in M. tuberculosis. Depletion of these branched chain amino acids in the medium led to loss of viability of the DeltailvB1 strain in vitro, resulting in a 4-log reduction in colony-forming units after 10 days. Survival kinetics of the mutant strain cultured in macrophages maintained with sub-optimal concentrations of the branched-chain amino acids did not show any loss of viability, indicating either that the intracellular environment was rich in these amino acids or that the other AHAS catalytic subunits were functional under these conditions. Furthermore, the growth kinetics of the DeltailvB1 strain in mice indicated that although this mutant strain showed defective growth in vivo, it could persist in the infected mice for a long time, and therefore could be a potential vaccine candidate. PMID- 19542001 TI - Pet secretion, internalization and induction of cell death during infection of epithelial cells by enteroaggregative Escherichia coli. AB - In an in vitro model using HEp-2 cells treated with purified plasmid-encoded toxin (Pet), we have identified morphological changes characterized by cell rounding and detachment after toxin internalization; these changes progress to cell death. However, these effects have not yet been shown to occur during the infection of epithelial cells by enteroaggregative Escherichia coli (EAEC). Here, we show that the secretion of Pet by EAEC is regulated at the transcriptional level, since secretion was inhibited in eukaryotic cell culture medium, although Pet was efficiently secreted in the same medium supplemented with tryptone. Inefficient secretion of Pet by EAEC in DMEM prevented cell detachment, whereas efficient Pet secretion in DMEM/tryptone increased cell detachment in a HEp-2 cell adherence assay. Interestingly, Pet toxin was efficiently delivered to epithelial cells, since it was internalized into epithelial cells infected with EAEC at similar concentrations to those obtained by using 37 microg ml(-1) purified Pet protein. Additionally, Pet was not internalized when the epithelial cells were infected with a pet clone, HB101(pCEFN1), unlike the wild-type strain, which has a high adherence capability. There is a correlation between Pet secretion by EAEC, the internalization of Pet into epithelial cells, cell detachment and cell death in EAEC-infected cells. The ratio between live and dead cells decreased in cells treated with wild-type EAEC in comparison with cells treated with an isogenic mutant in the pet gene, whereas the effects were restored by complementing the mutant with the pet gene. All these data indicate that Pet is an important virulence factor in the pathogenesis of EAEC infection. PMID- 19542002 TI - Interactions of Burkholderia cenocepacia and other Burkholderia cepacia complex bacteria with epithelial and phagocytic cells. AB - Burkholderia cenocepacia is a member of the B. cepacia complex (Bcc), a group of opportunistic bacteria that infect the airways of patients with cystic fibrosis (CF) and are extraordinarily resistant to almost all clinically useful antibiotics. Infections in CF patients with Bcc bacteria generally lead to a more rapid decline in lung function, and in some cases to the 'cepacia syndrome', a virtually deadly exacerbation of the lung infection with systemic manifestations. These characteristics of Bcc bacteria contribute to higher morbidity and mortality in infected CF patients. In the last 10 years considerable progress has been made in understanding the interactions between Bcc bacteria and mammalian host cells. Bcc isolates can survive either intracellularly within eukaryotic cells or extracellularly in host tissues. They survive within phagocytes and respiratory epithelial cells, and they have the ability to breach the respiratory epithelium layer. Survival and persistence of Bcc bacteria within host cells and tissues are believed to play a key role in pulmonary infection and to contribute to the persistent inflammation observed in patients with CF. This review summarizes recent findings concerning the interaction between Bcc bacteria and epithelial and phagocytic cells. PMID- 19542003 TI - Streptomyces roseoverticillatus produces two different poly(amino acid)s: lariat shaped gamma-poly(L-glutamic acid) and epsilon-poly(L-lysine). AB - The poly(amino acid)s gamma-poly(dl-glutamic acid) (gPGA) and epsilon-poly(l lysine) (ePL) are known to be natural linear poly(amino acid)s secreted by Bacillus spp. and Streptomyces spp., respectively. In this study, a Streptomyces strain producing both ePL and gPGA was identified. Mass spectrometry and other analyses revealed that the gPGA is a mixture of oligomers consisting of 10-13 l glutamic acid residues linked by isopeptide bonds. In contrast to the known Bacillus gPGA, the glutamic acid oligomers have a cyclodehydrated structure in each molecule. We previously reported that the ePL molecules secreted by the same Streptomyces strain disperse only slightly in an agar culture plate, as though they were larger molecules. This phenomenon is explicable by the observed polyion complex formation between the glutamic acid oligomers and ePLs. The glutamic acid oligomers control the ePL's dispersion, which would also affect the spatial distribution of the ePL's antimicrobial activity. Therefore, gene clustering or common use of the gene was presumed for biosynthesis of the two poly(amino acid)s. However, no gene for biosynthesis of the glutamic acid oligomer was found in the neighbouring region of that for ePL biosynthesis, and the glutamic acid oligomer was produced by a mutant in which the ePL biosynthetic gene was inactivated by gene disruption. PMID- 19542004 TI - Multiple redundant stress resistance mechanisms are induced in Salmonella enterica serovar Typhimurium in response to alteration of the intracellular environment via TLR4 signalling. AB - Toll-like receptor 4 (TLR4) senses bacterial LPS and is required for the control of systemic Salmonella enterica serovar Typhimurium infection in mice. The mechanisms of TLR4 activation and its downstream signalling cascades are well described, yet the direct effects on the pathogen of signalling via this receptor remain unknown. To investigate this we used microarray-based transcriptome profiling of intracellular S. Typhimurium during infection of primary bone marrow derived macrophages from wild-type and TLR4-deficient mice. We identified 17 S. Typhimurium genes that were upregulated in the presence of functional TLR4. Nine of these genes have putative functions in oxidative stress resistance. We therefore examined S. Typhimurium gene expression during infection of NADPH oxidase-deficient macrophages, which lack normal oxidative killing mechanisms. We identified significant overlap between the 'TLR4-responsive' and 'NADPH oxidase responsive' genes. This is new evidence for a link between TLR4 signalling and NADPH oxidase activity. Interestingly, with the exception of a dps mutant, S. Typhimurium strains lacking individual TLR4- and/or oxidative stress-responsive genes were not attenuated during intravenous murine infections. Our study shows that TLR4 activity, either directly or indirectly, induces the expression of multiple stress resistance genes during the intracellular life of S. Typhimurium. PMID- 19542005 TI - DNA polymerase X from Deinococcus radiodurans implicated in bacterial tolerance to DNA damage is characterized as a short patch base excision repair polymerase. AB - The Deinococcus radiodurans R1 genome encodes an X-family DNA repair polymerase homologous to eukaryotic DNA polymerase beta. The recombinant deinococcal polymerase X (PolX) purified from transgenic Escherichia coli showed deoxynucleotidyltransferase activity. Unlike the Klenow fragment of E. coli, this enzyme showed short patch DNA synthesis activity on heteropolymeric DNA substrate. The recombinant enzyme showed 5'-deoxyribose phosphate (5'-dRP) lyase activity and base excision repair function in vitro, with the help of externally supplied glycosylase and AP endonuclease functions. A polX disruption mutant of D. radiodurans expressing 5'-dRP lyase and a truncated polymerase domain was comparatively less sensitive to gamma-radiation than a polX deletion mutant. Both mutants showed higher sensitivity to hydrogen peroxide. Excision repair mutants of E. coli expressing this polymerase showed functional complementation of UV sensitivity. These results suggest the involvement of deinococcal polymerase X in DNA-damage tolerance of D. radiodurans, possibly by contributing to DNA double strand break repair and base excision repair. PMID- 19542006 TI - Identification and characterization of a family of toxin-antitoxin systems related to the Enterococcus faecalis plasmid pAD1 par addiction module. AB - The par locus of the Enterococcus faecalis plasmid pAD1 is an RNA-regulated addiction module encoding the peptide toxin Fst. Homology searches revealed that Fst belongs to a family of at least nine related peptides encoded on the chromosomes and plasmids of six different Gram-positive bacterial species. Comparison of an alignment of these peptides with the results of a saturation mutagenesis analysis indicated regions of the peptides important for biological function. Examination of the genetic context of the fst genes revealed that all of these peptides are encoded within par-like loci with conserved features similar to pAD1 par. All four Ent. faecalis family members were demonstrated to produce the expected toxin-encoding and regulatory RNA products. The locus from the Ent. faecalis plasmid pAMS1 was demonstrated to function as an addiction module and Fst was shown to be toxic to Staphylococcus aureus, suggesting that a plasmid-encoded module in that species is performing the same function. Thus, the pAD1-encoded par locus appears to be the prototype of a family of related loci found in several Gram-positive species. PMID- 19542007 TI - Illumination stimulates cAMP receptor protein-dependent transcriptional activation from regulatory regions containing class I and class II promoter elements in Synechocystis sp. PCC 6803. AB - The cAMP receptor protein (Crp) is a global transcriptional regulator that binds sequence-specific promoter elements when associated with cAMP. In the motile cyanobacterium Synechocystis sp. strain PCC 6803, intracellular cAMP increases when dark-adapted cells are illuminated. Previous work has established that Crp binds proposed Crp target sites upstream of slr1351 (murF), sll1874 (chlA(II)), sll1708 (narL), slr0442 and sll1268 in vitro, and that slr0442 is downregulated in a crp mutant during photoautotrophic growth. To identify additional Crp target genes in Synechocystis, 11 different Crp binding sites proposed during a previous computational survey were tested for in vitro sequence-specific binding and crp dependent transcription. The results indicate that murF, chlA(II) and slr0442 can be added as 'target genes of Sycrp1' in Synechocystis. Promoter mapping of the targets revealed the same close association of RNA polymerase and Crp as that found in Escherichia coli class I and class II Crp-regulated promoters, thereby strongly suggesting similar mechanisms of transcriptional activation. PMID- 19542008 TI - Uptake of the fluorescent probe FM4-64 by hyphae and haemolymph-derived in vivo hyphal bodies of the entomopathogenic fungus Beauveria bassiana. AB - The entomopathogenic fungus Beauveria bassiana is under intensive study as a pest biological control agent. B. bassiana produces several distinct single-cell types that include aerial conidia, in vitro blastospores and submerged conidia. Under appropriate nutrient conditions these cells can elaborate germ tubes that form hyphae, which in turn lead to the formation of a fungal mycelium. In addition, B. bassiana displays a dimorphic transition, producing in vivo specific yeast-like hyphal bodies during growth in the arthropod haemolymph. The amphiphilic styryl dye FM4-64 was used to investigate internalization and morphological features of in vitro and in vivo insect haemolymph-derived B. bassiana cells. In vitro blastospores and submerged conidia displayed a punctate pattern of internal labelling, whereas aerial conidia failed to internalize the dye under the conditions tested. FM4-64 was also taken up into both apical and subapical compartments of living hyphae in a time-dependent manner, with clearly observable vesicle labelling. Internalization, where occurring, was reversibly disrupted by lowering the temperature of the assay or by treatment with azide/fluoride and latrunculin A. Treatment with cytochalasin D and monensin also caused abnormal vesicle trafficking, although some staining of vesicles was noted. Fungal cells derived from infected Heliothis virescens haemolymph (in vivo cells) actively internalized FM4-64. The in vivo blastospores or hyphal bodies displayed bright membrane and internal vesicle staining, although diffuse staining of internal structures was also visible. These results suggest active uptake by different developmental stages of B. bassiana, including haemolymph-derived cells that can evade the insect immune system. PMID- 19542009 TI - The interaction between Listeria monocytogenes and the host gastrointestinal tract. AB - Listeria monocytogenes is a ubiquitous bacterium that causes significant foodborne disease with high mortality rates in immunocompromised adults. In pregnant women foodborne infection can give rise to infection of the fetus resulting in miscarriage. In addition, the bacterium has recently been demonstrated to cause localized gastrointestinal symptoms, predominantly in immunocompetent individuals. The murine model of systemic L. monocytogenes infection has provided numerous insights into the mechanisms of pathogenesis of this organism. However, recent application of transcriptomic and proteomic approaches as well as the development of new model systems has allowed a focus upon factors that influence adaptation to gastrointestinal environments and adhesion to and invasion of the gastrointestinal mucosa. In addition, the availability of a large number of complete L. monocytogenes genome sequences has permitted inter-strain comparisons and the identification of factors that may influence the emergence of 'epidemic' phenotypes. Here we review some of the exciting recent developments in the analysis of the interaction between L. monocytogenes and the host gastrointestinal tract. PMID- 19542010 TI - Burkholderia cenocepacia zinc metalloproteases influence resistance to antimicrobial peptides. AB - Burkholderia cenocepacia secretes two zinc-dependent metalloproteases, designated ZmpA and ZmpB. Previously, ZmpA and ZmpB have been shown to cleave several proteins important in host defence. In this study, the ability of ZmpA and ZmpB to digest and inactivate antimicrobial peptides involved in innate immunity was examined. ZmpB but not ZmpA cleaved beta-defensin-1. ZmpA but not ZmpB cleaved the cathelicidin LL-37. Both enzymes cleaved elafin and secretory leukocyte inhibitor, which are antimicrobial peptides as well as neutrophil elastase inhibitors. Both ZmpA and ZmpB cleaved protamine, a fish antimicrobial peptide, and a zmpA zmpB mutant was more sensitive to protamine killing than the parental strain. ZmpA or ZmpB cleavage of elafin inactivated its anti-protease activity. The effect of ZmpA and ZmpB on the neutrophil proteases elastase and cathepsin G was also examined but neither enzyme was active against these host proteases. These studies suggest that ZmpA and ZmpB may influence the resistance of B. cenocepacia to host antimicrobial peptides as well as alter the host protease/anti-protease balance in chronic respiratory infections. PMID- 19542011 TI - Effects of a diet higher in carbohydrate/lower in fat versus lower in carbohydrate/higher in monounsaturated fat on postmeal triglyceride concentrations and other cardiovascular risk factors in type 1 diabetes. AB - OBJECTIVE: To compare the effects of a eucaloric diet higher in carbohydrate/lower in fat versus lower in carbohydrate/higher in monounsaturated fat on postmeal triglyceride (TG) concentrations and other cardiovascular disease risk factors in nonobese subjects with type 1 diabetes and in good glycemic control. RESEARCH DESIGN AND METHODS: In a parallel group design study, 30 subjects were randomly assigned and completed one of the two eucaloric diets. Assessments included: BMI, blood pressure, A1C, plasma lipids, and markers of oxidation, thrombosis, and inflammation. At 6 months, subjects were hospitalized for 24 h to measure plasma TG excursions. RESULTS: There were no significant differences between groups other than decreased plasminogen activator inhibitor 1 (PAI-1) levels and weight gain in the lower-carbohydrate/higher-monounsaturated fat group. During the 24-h testing, the lower-carbohydrate/higher-monounsaturated fat group had a lower plasma TG profile. CONCLUSIONS: A diet lower in carbohydrate/higher in monounsaturated fat could offer an appropriate choice for nonobese type 1 diabetic individuals with good metabolic and weight control. PMID- 19542012 TI - Effects of a protein preload on gastric emptying, glycemia, and gut hormones after a carbohydrate meal in diet-controlled type 2 diabetes. AB - OBJECTIVE: We evaluated whether a whey preload could slow gastric emptying, stimulate incretin hormones, and attenuate postprandial glycemia in type 2 diabetes. RESEARCH DESIGN AND METHODS: Eight type 2 diabetic patients ingested 350 ml beef soup 30 min before a potato meal; 55 g whey was added to either the soup (whey preload) or potato (whey in meal) or no whey was given. RESULTS: Gastric emptying was slowest after the whey preload (P < 0.0005). The incremental area under the blood glucose curve was less after the whey preload and whey in meal than after no whey (P < 0.005). Plasma glucose-dependent insulinotropic polypeptide, insulin, and cholecystokinin concentrations were higher on both whey days than after no whey, whereas glucagon-like peptide 1 was greatest after the whey preload (P < 0.05). CONCLUSIONS: Whey protein consumed before a carbohydrate meal can stimulate insulin and incretin hormone secretion and slow gastric emptying, leading to marked reduction in postprandial glycemia in type 2 diabetes. PMID- 19542013 TI - Reversible oxidative modification: a key mechanism of Na+-K+ pump regulation. AB - Angiotensin II (Ang II) inhibits the cardiac sarcolemmal Na(+)-K(+) pump via protein kinase (PK)C-dependent activation of NADPH oxidase. We examined whether this is mediated by oxidative modification of the pump subunits. We detected glutathionylation of beta(1), but not alpha(1), subunits in rabbit ventricular myocytes at baseline. beta(1) Subunit glutathionylation was increased by peroxynitrite (ONOO(-)), paraquat, or activation of NADPH oxidase by Ang II. Increased glutathionylation was associated with decreased alpha(1)/beta(1) subunit coimmunoprecipitation. Glutathionylation was reversed after addition of superoxide dismutase. Glutaredoxin 1, which catalyzes deglutathionylation, coimmunoprecipitated with beta(1) subunit and, when included in patch pipette solutions, abolished paraquat-induced inhibition of myocyte Na(+)-K(+) pump current (I(p)). Cysteine (Cys46) of the beta(1) subunit was the likely candidate for glutathionylation. We expressed Na(+)-K(+) pump alpha(1) subunits with wild type or Cys46-mutated beta(1) subunits in Xenopus oocytes. ONOO(-) induced glutathionylation of beta(1) subunit and a decrease in Na(+)-K(+) pump turnover number. This was eliminated by mutation of Cys46. ONOO(-) also induced glutathionylation of the Na(+)-K(+) ATPase beta(1) subunit from pig kidney. This was associated with a approximately 2-fold decrease in the rate-limiting E(2)- >E(1) conformational change of the pump, as determined by RH421 fluorescence. We propose that kinase-dependent regulation of the Na(+)-K(+) pump occurs via glutathionylation of its beta(1) subunit at Cys46. These findings have implications for pathophysiological conditions characterized by neurohormonal dysregulation, myocardial oxidative stress and raised myocyte Na(+) levels. PMID- 19542014 TI - MicroRNA-145, a novel smooth muscle cell phenotypic marker and modulator, controls vascular neointimal lesion formation. AB - Phenotypic modulation of vascular smooth muscle cells (VSMCs) plays a critical role in the pathogenesis of a variety of proliferative vascular diseases. Recently, we have found that microRNA (miRNA) miR-145 is the most abundant miRNA in normal vascular walls and in freshly isolated VSMCs; however, the role of miR 145 in VSMC phenotypic modulation and vascular diseases is currently unknown. Here we find that miR-145 is selectively expressed in VSMCs of the vascular wall and its expression is significantly downregulated in the vascular walls with neointimal lesion formation and in cultured dedifferentiated VSMCs. More importantly, both in cultured rat VSMCs in vitro and in balloon-injured rat carotid arteries in vivo, we demonstrate that the noncoding RNA miR-145 is a novel phenotypic marker and a novel phenotypic modulator of VSMCs. VSMC differentiation marker genes such as SM alpha-actin, calponin, and SM-MHC are upregulated by premiR-145 or adenovirus expressing miR-145 (Ad-miR-145) but are downregulated by the miR-145 inhibitor 2'OMe-miR-145. We have further identified that miR-145-mediated phenotypic modulation of VSMCs is through its target gene KLF5 and its downstream signaling molecule, myocardin. Finally, restoration of miR-145 in balloon-injured arteries via Ad-miR-145 inhibits neointimal growth. We conclude that miR-145 is a novel VSMC phenotypic marker and modulator that is able of controlling vascular neointimal lesion formation. These novel findings may have extensive implications for the diagnosis and therapy of a variety of proliferative vascular diseases. PMID- 19542016 TI - An unbiased chemical biology screen identifies agents that modulate uptake of oxidized LDL by macrophages. AB - Macrophage-derived foam cells are thought to play a major role in atherosclerotic lesion formation and progression. An automated assay was established to evaluate the uptake of fluorescently labeled oxidized low-density lipoprotein (oxLDL) by a monocyte/macrophage cell line. The assay was used to screen 480 known bioactive compounds. Twenty-two active compounds were identified. Efficacy studies in peritoneal macrophages demonstrated a high rate of concordance with the initial screening results. Inhibitory compounds confirmed important previous findings and identified new drugs of interest including: 3 blockers of nuclear factor kappab activation, 2 protein kinase C inhibitors, a phospholipase C inhibitor, and 2 antipsychotic drugs. In addition, an opioid receptor agonist was found to increase the oxLDL uptake of macrophages. The involvement of nuclear factor kappaB in oxLDL uptake was validated in peritoneal macrophages in vivo. The results support a model in which oxLDL uptake is dependent on the activation of multiple intracellular signaling pathways that culminate in actin-mediated lipoprotein internalization. PMID- 19542015 TI - Angiomodulin is a specific marker of vasculature and regulates vascular endothelial growth factor-A-dependent neoangiogenesis. AB - Blood vessel formation is controlled by the balance between pro- and antiangiogenic pathways. Although much is known about the factors that drive sprouting of neovessels, the factors that stabilize and pattern neovessels are undefined. The expression of angiomodulin (AGM), a vascular endothelial growth factor (VEGF)-A binding protein, was increased in the vasculature of several human tumors as compared to normal tissue, raising the hypothesis that AGM may modulate VEGF-A-dependent vascular patterning. To elucidate the expression pattern of AGM, we developed an AGM knockin reporter mouse (AGM(lacZ/+)), with which we demonstrate that AGM is predominantly expressed in the vasculature of developing embryos and adult organs. During physiological and pathological angiogenesis, AGM is upregulated in the angiogenic vasculature. Using the zebrafish model, we found that AGM is restricted to developing vasculature by 17 to 22 hours postfertilization. Blockade of AGM activity with morpholino oligomers results in prominent angiogenesis defects in vascular sprouting and remodeling. Concurrent knockdown of both AGM and VEGF-A results in synergistic angiogenesis defects. When VEGF-A is overexpressed, the compensatory induction of the VEGF-A receptor, VEGFR2/flk-1, is blocked by the simultaneous injection of AGM morpholino oligomers. These results demonstrate that the vascular-specific marker AGM modulates vascular remodeling in part by temporizing the proangiogenic effects of VEGF-A. PMID- 19542018 TI - Add some fat to vascular progenitor cell therapy. PMID- 19542017 TI - Dysregulation of positive transcription elongation factor B and myocardial hypertrophy. PMID- 19542019 TI - Somitovasculin, a novel endothelial-specific transcript involved in the vasculature development. AB - OBJECTIVE: We recently isolated and characterized endothelial-like CD31(+) cells derived from mouse embryonic stem (mES) cells and identified their transcriptome. The main objective of this study was to determine the functional relevance of the transcripts of unknown function (TUF) for vasculature development. METHODS AND RESULTS: We selected 2 TUFs of more than 27 to study their role for blood vessel development in zebrafish. Morpholino (MO) knockdown of the zebrafish orthologs of the first TUF (TUF1, mouse cDNA BC022623) showed disruption of the intersegmental vessels (ISV) at 2 days postfertilization as observed by live imaging of fli:EGFP transgenic embryos. The morphants showed abnormal blood circulation, but no effect on hematopoiesis was observed as demonstrated by gata-1 in situ hybridizations. Because knockdown of TUF1 resulted in disruption of the ISV patterning we named the TUF1 somitovasculin. TUF2 has been identified as cDNA clone BC020535. The MO knockdown of TUF2 resulted in a phenotype with an enlarged heart and the embryos lacked circulation completely. CONCLUSIONS: We have shown the participation of a novel transcript (named somitovasculin) in circulatory vessel development. The combination of expression profiling in differentiating mES cells and the zebrafish model has the potential for rapid identification and functional characterization of TUFs. PMID- 19542020 TI - Is diabetes mellitus an independent risk factor for venous thromboembolism?: a population-based case-control study. AB - OBJECTIVE: Although diabetes mellitus is reported as a risk factor for venous thromboembolism (VTE), persons with diabetes are frequently hospitalized for medical illness or surgery, or confined to a nursing home, all major VTE risk factors. Consequently, we tested diabetes for an independent association with incident VTE. METHODS AND RESULTS: Using Rochester Epidemiology Project resources, we identified all Olmsted County, Minn residents who met objective criteria for incident VTE over the 25-year period, 1976 to 2000 (n=1922), and 1 to 2 resident controls per case, matched on age, gender, and length of medical history (n=2115). Complete medical histories in the community were reviewed for previously identified independent VTE risk factors and diabetes-related variables. We tested diabetes and diabetes complications (retinopathy, nephropathy or neuropathy, and ketoacidosis) as potential VTE risk factors, both alone and after adjusting for VTE risk factors. Univariately, diabetes by clinical diagnosis or by stricter criteria (fasting ambulatory blood glucose >/=140 mg/dL or antidiabetic drug therapy), and diabetes complications, were associated with overall incident VTE. However, after controlling for hospitalization for major surgery or medical illness and nursing home confinement, diabetes was no longer associated with VTE. CONCLUSIONS: Diabetes mellitus and diabetes complications are not independent risk factors for incident VTE. PMID- 19542021 TI - Activation of NF-kappaB by palmitate in endothelial cells: a key role for NADPH oxidase-derived superoxide in response to TLR4 activation. AB - OBJECTIVE: We investigated whether NADPH oxidase-dependent production of superoxide contributes to activation of NF-kappaB in endothelial cells by the saturated free fatty acid palmitate. METHODS AND RESULTS: After incubation of human endothelial cells with palmitate at a concentration known to induce cellular inflammation (100 mumol/L), we measured superoxide levels by using electron spin resonance spectroscopy and the spin trap 1-hydroxy-3 methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine (CMH). Palmitate exposure induced a >2-fold increase in superoxide levels, an effect associated with activation of NF-kappaB signaling as measured by phospho-IkappaBalpha, NF-kappaB activity, IL 6, and ICAM expression. Reduction in superoxide levels by each of 3 different interventions-pretreatment with superoxide dismutase (SOD), diphenylene iodinium (DPI), or knockdown of NADPH oxidase 4 (NOX4) by siRNA-attenuated palmitate mediated NF-kappaB signaling. Inhibition of toll like receptor-4 (TLR4) signaling also suppressed palmitate-mediated superoxide production and associated inflammation, whereas palmitate-mediated superoxide production was not affected by overexpression of a phosphorylation mutant IkappaBalpha (NF-kappaB super repressor) that blocks cellular inflammation downstream of IKKbeta/NF-kappaB. Finally, high-fat feeding increased expression of NOX4 and an upstream activator, bone morphogenic protein (BMP4), in thoracic aortic tissue from C57BL/6 mice, but not in TLR4(-/-) mice, compared to low-fat fed controls. CONCLUSIONS: These results suggest that NADPH oxidase-dependent superoxide production links palmitate-stimulated TLR4 activation to NF-kappaB signaling in endothelial cells. PMID- 19542022 TI - Stimulation of coronary collateral growth by granulocyte stimulating factor: role of reactive oxygen species. AB - OBJECTIVE: The purpose of this study was to determine whether G-CSF promotes coronary collateral growth (CCG) and decipher the mechanism for this stimulation. METHODS AND RESULTS: In a rat model of repetitive episodic myocardial ischemia (RI, 40 seconds LAD occlusion every 20 minutes for 2 hours and 20 minutes, 3 times/d for 5 days) CCG was deduced from collateral-dependent flow (flow to LAD region during occlusion). After RI, G-CSF (100 microg/kg/d) increased CCG (P<0.01) (0.47+/-0.15) versus vehicle (0.14+/-0.06). Surprisingly, G-CSF treatment without RI increased CCG (0.57+/-0.18) equal to G-CSF+RI. We evaluated ROS by dihydroethidine (DHE) fluorescence (LV injection, 60 microg/kg, during two episodes of ischemia). DHE fluorescence was double in G-CSF+RI versus vehicle+RI (P<0.01), and even higher in G-CSF without RI (P<0.01). Interestingly, the DHE signal did not colocalize with myeloperoxidase (immunostaining, neutrophil marker) but appeared in cardiac myocytes. The study of isolated cardiac myocytes revealed the cytokine stimulates ROS which elicit production of angiogenic factors. Apocynin inhibited G-CSF effects both in vivo and in vitro. CONCLUSIONS: G-CSF stimulates ROS production directly in cardiomyocytes, which plays a pivotal role in triggering adaptations of the heart to ischemia including growth of the coronary collaterals. PMID- 19542023 TI - Targeted metabolomic evaluation of arginine methylation and cardiovascular risks: potential mechanisms beyond nitric oxide synthase inhibition. AB - BACKGROUND: We examine the relationship of related posttranslational modification products of arginine methylation and coronary artery disease (CAD) phenotypes. METHODS AND RESULTS: Plasma was isolated from 1011 consecutive consenting subjects undergoing elective diagnostic cardiac catheterization, and future major adverse cardiac events (MACE, including myocardial infarction, stroke, and death) at 3 years were investigated. Plasma levels of asymmetrical dimethylarginine (ADMA, endogenous nitric oxide synthase [NOS] inhibitor), symmetrical dimethylarginine (SDMA, which lacks NOS inhibitory activity), N-mono methylarginine (MMA, a potent NOS inhibitor), methyl-lysine (Methyl-Lys, an unrelated methylated amino acid), arginine, and its major catabolites (citrulline and ornithine) were quantified simultaneously by stable isotope dilution HPLC with online electrospray ionization tandem mass spectrometry and adjusted for traditional risk factors, C-reactive protein, and estimated creatinine clearance. High SDMA levels (adjusted odds ratio [OR] 1.6, 95%CI, 1.1 to 2.6, P<0.001), low MMA (adjusted OR 0.5, 95%CI 0.4 to 0.8, P=0.007), but not ADMA (adjusted OR 1.3, 95%CI 0.88 to 2.0, P=0.177) were associated with increased prevalence of significantly obstructive CAD. Elevated levels of SDMA (adjusted Hazard Ratio [HR] 2.4, 95%CI 1.2 to 4.6, P=0.009), ADMA (adjusted HR 2.2, 95%CI 1.2 to 4.0, P=0.015), as well as an integrated index of arginine methylation [ArgMI=(ADMA+SDMA)/MMA] (adjusted HR 2.4, 95%CI 1.3 to 4.5, P=0.006) were significant independent predictors of incident MACE. ArgMI was predictive of incident MACE even following adjustments for global arginine bioavailability, particularly within secondary prevention patients. CONCLUSIONS: ADMA, SDMA, and the integrated quantification of arginine methylation (in the form of a methylation index) provided independent risk prediction for both significantly obstructive CAD and incident MACE in stable patients undergoing cardiac evaluation. These results suggest that factors beyond direct NOS inhibition contribute to the clinical associations between methylarginines and CAD outcomes. PMID- 19542024 TI - Heme oxygenase-1 increases endothelial progenitor cells. AB - OBJECTIVE: Induction of heme oxygenase-1 (HO-1) protects against atherosclerotic disease in part by promoting reendothelialization. As endothelial progenitor cells (EPCs) contribute to reendothelialization, we examined the role of HO-1 on bone marrow and circulating EPCs. METHODS AND RESULTS: In a rabbit model of aortic balloon injury, pharmacological induction of HO-1 enhanced reendothelialization at sites with and without adjacent blood vessels, the latter indicative of a contribution by EPCs. Coinciding with maximal HO-1 induction in the injured vessel, plasma concentrations of bilirubin and the numbers of circulating progenitor cells were elevated. Both processes were abolished by cotreatment of the animals with an inhibitor of HO-1. Inducers of HO-1 promoted bone marrow cells to form progenitor cell colonies, and Flk1(+)/Sca-1(+)-cells to adhere to the luminal surface of the injured vessel. In noninjured mice, HO-1 inducers also increased bone marrow and circulating EPCs, and the ability of these cells to differentiate and form colonies. Compared to wild-type mice, bone marrow cells from HO-1(-/-) mice generated fewer endothelial colony-forming cells, and HO-1 inducers failed to promote CFU-Hill colony formation. CONCLUSIONS: These findings suggest that HO-1 contributes to vascular repair by increasing circulating EPCs derived from the bone marrow. PMID- 19542025 TI - Political economy of US states and rates of fatal occupational injury. AB - OBJECTIVES: We investigated the extent to which the political economy of US states, including the relative power of organized labor, predicts rates of fatal occupational injury. METHODS: We described states' political economies with 6 contextual variables measuring social and political conditions: "right-to-work" laws, union membership density, labor grievance rates, state government debt, unemployment rates, and social wage payments. We obtained data on fatal occupational injuries from the National Traumatic Occupational Fatality surveillance system and population data from the US national census. We used Poisson regression methods to analyze relationships for the years 1980 and 1995. RESULTS: States differed notably with respect to political-economic characteristics and occupational fatality rates, although these characteristics were more homogeneous within rather than between regions. Industry and workforce composition contributed significantly to differences in state injury rates, but political-economic characteristics of states were also significantly associated with injury rates, after adjustment accounting for those factors. CONCLUSIONS: Higher rates of fatal occupational injury were associated with a state policy climate favoring business over labor, with distinct regional clustering of such state policies in the South and Northeast. PMID- 19542026 TI - Social conditions and health. PMID- 19542027 TI - Indoor residual spraying of DDT for malaria control. PMID- 19542028 TI - Smoking affects womens' sex hormone-regulated body form. PMID- 19542029 TI - Carrier detection and clinical uncertainty: the case for public health ethics. PMID- 19542030 TI - Clinical and ethical considerations in managing carrier detection. PMID- 19542031 TI - AJPH endorsement of transparency, clarity, and rigor. PMID- 19542032 TI - Promoting health in American-occupied Japan. Resistance to allied public health measures, 1945-1952. AB - As soon as the authority of the Public Health and Welfare Section (PHW) of the Supreme Commander for Allied Powers waned in May 1951, the Japanese government overturned several measures it had implemented. Although the PHW contributed greatly toward improving public health conditions, not all of its activities were models of cooperative success. Many Japanese perceived some measures-terminated pensions for wounded Japanese veterans, lack of support for segregated orphanages for mixed-race children, and suppression of Japanese atomic bomb medical reports as promoting US national interest at the expense of Japanese public health needs. Similarly, the PHW's upgrade of nursing education and separation of the professions of medicine and pharmacy were reversed because neither professionals nor the public saw these measures as urgent. Their reinstitution toward the end of the twentieth century suggests that the progressive measures were sound, but broke too sharply with Japanese tradition and were enforced prematurely. PMID- 19542033 TI - The limits of collaboration: a qualitative study of community ethical review of environmental health research. AB - OBJECTIVES: We assessed the effectiveness of various systems of community participation in ethical review of environmental health research. METHODS: We used situation analysis methods and a global workspace theoretical framework to conduct comparative case studies of 3 research organizations at 1 medical center. RESULTS: We found a general institutional commitment to community review as well as personal commitment from some participants in the process. However, difficulty in communicating across divides of knowledge and privilege created serious gaps in implementation, leaving research vulnerable to validity threats (such as misinterpretation of findings) and communities vulnerable to harm. The methods used in each collaboration solved some, but not all, of the problems that hindered communication. CONCLUSIONS: Researchers, community spokespersons, and institutional review boards constitute organizational groups with strong internal ties and highly developed cultures. Few cross-linkages and little knowledge of each other cause significant distortion of information and other forms of miscommunication between groups. Our data suggest that organizations designed to protect human volunteers are in the best position to take the lead in implementing community review. PMID- 19542034 TI - Effects of a prekindergarten educational intervention on adult health: 37-year follow-up results of a randomized controlled trial. AB - OBJECTIVES: We used 37 years of follow-up data from a randomized controlled trial to explore the linkage between an early educational intervention and adult health. METHODS: We analyzed data from the High/Scope Perry Preschool Program (PPP), an early school-based intervention in which 123 children were randomized to a prekindergarten education group or a control group. In addition to exploring the effects of the program on health behavioral risk factors and health outcomes, we examined the extent to which educational attainment, income, family environment, and health insurance access mediated the relationship between randomization to PPP and behavioral and health outcomes. RESULTS: The PPP led to improvements in educational attainment, health insurance, income, and family environment Improvements in these domains, in turn, lead to improvements in an array of behavioral risk factors and health (P = .01). However, despite these reductions in behavioral risk factors, participants did not exhibit any overall improvement in physical health outcomes by the age of 40 years. CONCLUSIONS: Early education reduces health behavioral risk factors by enhancing educational attainment, health insurance coverage, income, and family environments. Further follow-up will be needed to determine the long-term health effects of PPP. PMID- 19542035 TI - Prevalence, characteristics, and associated health and health care of family homelessness among fifth-grade students. AB - OBJECTIVES: We describe the lifetime prevalence and associated health-related concerns of family homelessness among fifth-grade students. METHODS: We used a population-based, cross-sectional survey of 5147 fifth-grade students in 3 US cities to analyze parent-reported measures of family homelessness, child health status, health care access and use, and emotional, developmental, and behavioral health and child-reported measures of health-related quality of life and exposure to violence. RESULTS: Seven percent of parents reported that they and their child had experienced homelessness (i.e., staying in shelters, cars, or on the street). Black children and children in the poorest families had the highest prevalence of homelessness (11%). In adjusted analyses, most general health measures were similar for children who had and had not been homeless. Children who had ever experienced homelessness were more likely to have an emotional, behavioral, or developmental problem (odds ratio [OR] = 1.7; 95% confidence interval [CI] = 1.1, 2.6; P = .01), to have received mental health care (OR = 2.2; 95% CI = 1.6, 3.2; P < .001), and to have witnessed serious violence with a knife (OR = 1.6; 95% CI = 1.1, 2.3; P = .007) than were children who were never homeless. CONCLUSIONS: Family homelessness affects a substantial minority of fifth-grade children and may have an impact on their emotional, developmental, and behavioral health. PMID- 19542036 TI - Secondhand smoke in Pennsylvania casinos: a study of nonsmokers' exposure, dose, and risk. AB - OBJECTIVES: I assessed air pollution, ventilation, and nonsmokers' risk from secondhand smoke (SHS) in Pennsylvania casinos exempted from a statewide smoke free workplace law. METHODS: I measured respirable suspended particles (RSPs), particulate polycyclic aromatic hydrocarbons (PPAHs), and carbon dioxide inside and outside casinos; measured changes in patrons' urine cotinine after casino visits; and assessed SHS impact on workers and patrons, using exposure-response models, air quality standards, and odor and irritation thresholds. RESULTS: PPAH and RSP concentrations in casinos were, on average, 4 and 6 times, respectively, that of outdoor levels despite generous ventilation and low smoking prevalence. SHS infiltrated into nonsmoking gaming areas. Patrons' urine cotinine increased 1.9 ng/mL on average after about 4-hour visits. CONCLUSIONS: SHS-induced heart disease and lung cancer will cause an estimated 6 Pennsylvania casino workers' deaths annually per 10,000 at risk, 5-fold the death rate from Pennsylvania mining disasters. Casinos should not be exempt from smoke-free workplace laws. PMID- 19542038 TI - Pathways to and from homelessness and associated psychosocial outcomes among adolescents leaving the foster care system. AB - OBJECTIVES: We evaluated the prevalence and nature of housing problems among adolescents leaving foster care because of their age to provide evidence that can inform public and programmatic policies designed to prevent homelessness. METHODS: Housing and psychosocial outcomes in a sample of 265 adolescents who left the foster care system in 2002 and 2003 in a large midwestern metropolitan area were evaluated over a 2-year follow-up period. Analyses focused on identifying latent housing trajectory categories across the first 2 years after participants' exit from foster care. RESULTS: Findings revealed 4 latent housing classifications. Most participants (57%) had experienced stable housing situations since their exit from foster care. Those in the remaining 3 categories endured housing problems, and 20% were chronically homeless during the follow-up period. Housing instability was related to emotional and behavioral problems, physical and sexual victimization, criminal conviction, and high school dropout. CONCLUSIONS: Adolescents in foster care are at considerable risk of homelessness. Preventive initiatives can reduce homelessness in this population by implementing improved foster care programming and developing empirically informed interventions targeting foster care adolescents. PMID- 19542037 TI - Use of a social and character development program to prevent substance use, violent behaviors, and sexual activity among elementary-school students in Hawaii. AB - OBJECTIVES: We assessed the effectiveness of a 5-year trial of a comprehensive school-based program designed to prevent substance use, violent behaviors, and sexual activity among elementary-school students. METHODS: We used a matched pair, cluster-randomized, controlled design, with 10 intervention schools and 10 control schools. Fifth-graders (N = 1714) self-reported on lifetime substance use, violence, and voluntary sexual activity. Teachers of participant students reported on student (N = 1225) substance use and violence. RESULTS: Two-level random-effects count models (with students nested within schools) indicated that student-reported substance use (rate ratio [RR] = 0.41; 90% confidence interval [CI] = 0.25, 0.66) and violence (RR = 0.42; 90% CI = 0.24, 0.73) were significantly lower for students attending intervention schools. A 2-level random effects binary model indicated that sexual activity was lower (odds ratio = 0.24; 90% CI = 0.08, 0.66) for intervention students. Teacher reports substantiated the effects seen for student-reported data. Dose-response analyses indicated that students exposed to the program for at least 3 years had significantly lower rates of all negative behaviors. CONCLUSIONS: Risk-related behaviors were substantially reduced for students who participated in the program, providing evidence that a comprehensive school-based program can have a strong beneficial effect on student behavior. PMID- 19542039 TI - Using sickness absence records to predict future depression in a working population: prospective findings from the GAZEL cohort. AB - OBJECTIVES: We tested the hypothesis that sickness absence from work predicts workers' risk of later depression. METHODS: Study participants (n = 7391) belonged to the French GAZEL cohort of employees of the national gas and electricity company. Sickness absence data (1996-1999) were obtained from company records. Participants' depression in 1996 and 1999 was assessed with the Center for Epidemiologic Studies-Depression (CES-D) scale. The analyses were controlled for baseline age, gender, marital status, occupational grade, tobacco smoking status, alcohol consumption, subthreshold depressive symptoms, and work stress. RESULTS: Among workers who were free of depression in 1996, 13% had depression in 1999. Compared with workers with no sickness absence during the study period, those with sickness absence were more likely to be depressed at follow-up (for 1 period of sickness absence, fully adjusted odds ratio [OR] = 1.53, 95% confidence interval [CI] = 1.28, 1.82; for 2 or more periods, fully adjusted OR = 1.95, 95% CI = 1.61, 2.36). Future depression was predicted both by psychiatric and nonpsychiatric sickness absence (fully adjusted OR = 3.79 [95% CI = 2.81, 5.10] and 1.41 [95% CI = 1.21, 1.65], respectively). CONCLUSIONS: Sickness absence records may help identify workers vulnerable to future depression. PMID- 19542040 TI - Influences of cross-border mobility on tuberculosis diagnoses and treatment interruption among injection drug users in Tijuana, Mexico. AB - OBJECTIVES: We sought to identify correlates of reported lifetime diagnoses of TB among injection drug users in the border city of Tijuana, Mexico. METHODS: Injection drug users in Tijuana were recruited into a prospective cohort study during 2006 and 2007. We used weighted multivariate logistic regression to identify correlates of TB diagnoses. RESULTS: Of the 1056 participants, 103 (9.8%) reported a history of TB, among whom 93% received anti-TB medication and 80% were diagnosed in the United States. Treatment was prematurely halted among 8% of patients; deportation from the United States was the cause of half of these treatment interruptions. History of travel to (odds ratio [OR] = 6.44; 95% confidence interval [CI] = 1.53, 27.20) or deportation from (OR = 1.83; 95% CI = 1.07, 3.12) the United States and incarceration (OR = 2.20; 95% CI = 1.06, 4.58) were independently associated with a reported lifetime diagnosis of TB. CONCLUSIONS: Mobility and migration are important factors in identifying and treating TB patients diagnosed in the US-Mexico border region. Strengthening capacity on both sides of the border to identify, monitor, and treat TB is a priority. PMID- 19542041 TI - Gender-specific correlates of incarceration among marginally housed individuals in San Francisco. AB - OBJECTIVES: We assessed how different patterns of housing instability affect incarceration and whether correlates of incarceration are gender specific. METHODS: We used multivariate logistic regression to assess associations between patterns of housing instability and recent jail stays among a reproducible sample of 1175 marginally housed adults in San Francisco, California. RESULTS: Over the previous year, 71% of men and 21% of women in the sample reported jail stays. Among women, long-term single-room occupancy hotel stays ( > 90 days) were protective for incarceration. Stays in the street were associated with incarceration among both genders, but among men, short-term (i.e., or =85 years old. Hospital charges totaled $1.51 billion; 24.0% ($362.5 million) of this total was attributable to racial disparities, 70.8% ($256.5 million) of which stemmed from the 36.6% of patients <65 years old. Most of the acute outcomes were poorer for AAs compared with CAs across age groups. CONCLUSIONS: Racial disparities in stroke admissions are more pronounced in younger age groups and result in significant economic consequences. Although AA stroke patients experienced generally worse acute outcomes than did CAs, these differences appear to be relatively consistent across age groups. PMID- 19542050 TI - Liver natural killer and natural killer T cells: immunobiology and emerging roles in liver diseases. AB - Hepatic lymphocytes are enriched in NK and NKT cells that play important roles in antiviral and antitumor defenses and in the pathogenesis of chronic liver disease. In this review, we discuss the differential distribution of NK and NKT cells in mouse, rat, and human livers, the ultrastructural similarities and differences between liver NK and NKT cells, and the regulation of liver NK and NKT cells in a variety of murine liver injury models. We also summarize recent findings about the role of NK and NKT cells in liver injury, fibrosis, and repair. In general, NK and NKT cells accelerate liver injury by producing proinflammatory cytokines and killing hepatocytes. NK cells inhibit liver fibrosis via killing early-activated and senescent-activated stellate cells and producing IFN-gamma. In regulating liver fibrosis, NKT cells appear to be less important than NK cells as a result of hepatic NKT cell tolerance. NK cells inhibit liver regeneration by producing IFN-gamma and killing hepatocytes; however, the role of NK cells on the proliferation of liver progenitor cells and the role of NKT cells in liver regeneration have been controversial. The emerging roles of NK/NKT cells in chronic human liver disease will also be discussed.Understanding the role of NK and NKT cells in the pathogenesis of chronic liver disease may help us design better therapies to treat patients with this disease. PMID- 19542055 TI - Cryptogenic stroke: cryptic definition? PMID- 19542056 TI - The impact of body mass index on mortality after stroke. AB - BACKGROUND AND PURPOSE: Little is known about the contribution of obesity to the higher mortality risk among stroke survivors. We assessed the independent association between body mass index (BMI) and mortality among stroke survivors. METHODS: Cross-sectional and prospective data from a nationally representative survey of noninstitutionalized civilian U.S. population aged 25 or older (n=20 050) with a baseline history of stroke (n=644) followed up from survey participation (1988-1994) through mortality assessment in 2000. Relationships between BMI and mortality attributable to all causes or cardiovascular causes were examined after adjusting for established prognosticators after stroke. RESULTS: Stroke survivors were more likely to be overweight (BMI 25 to 29 kg/m2) or obese (BMI > or =30 kg/m2 than those without stroke (64.3% versus 53.2%, P=0.003). In multivariable analysis, overall risk for all-cause mortality increased per kg/m2 of higher BMI (P=0.030), but an interaction between age and BMI (P=0.009) revealed that the association of higher BMI with mortality risk was strongest in younger individuals and declined linearly with increasing age, such that in the elderly, overweightness and obesity had a protective effect. The results were similar for the cardiovascular mortality outcome. CONCLUSIONS: Higher BMI after stroke is associated with a greater risk of all-cause and cardiovascular death among younger individuals. Younger stroke survivors may especially benefit from more vigorous efforts to monitor and treat obesity. PMID- 19542057 TI - Urgency of carotid endarterectomy for secondary stroke prevention: results from the Registry of the Canadian Stroke Network. AB - BACKGROUND AND PURPOSE: The benefit of carotid endarterectomy for preventing recurrent stroke is maximal when surgery is performed within 2 weeks after ischemic stroke or transient ischemic attack; the benefit is reduced when surgery is delayed >2 weeks and essentially lost if delayed >3 months. Guidelines recommend endarterectomy within 2 weeks poststroke/transient ischemic attack for patients with symptomatic carotid stenosis. This study examined time to endarterectomy at designated stroke centers as a measure of evidence-based best practices for stroke prevention. METHODS: From the Registry of the Canadian Stroke Network, we identified all consecutive patients presenting with acute ischemic stroke or transient ischemic attack at 12 provincial stroke centers (Ontario, Canada, 2003 to 2006) and selected those with unilateral symptomatic carotid stenosis of moderate (50% to 69%) or severe (70% to 99%) degree. Using linkages to administrative databases, we identified patients who underwent carotid endarterectomy within 6 months after the symptomatic event and calculated the time intervals between the index event and surgery. We compared the timing of surgery according to age, sex, degree of stenosis, index event, geographic region, and year. Logistic regression assessed variables associated with early surgery. RESULTS: One hundred five patients underwent endarterectomy for unilateral symptomatic carotid stenosis (50% to 99%) within 6 months of the index event. The median time from index event to surgery was 30 days (interquartile range, 10 to 81). Only one third (38 of 105) received endarterectomy within the recommended 2-week target timeframe, and in one fourth (26 of 105), surgery was delayed >3 months. Surgery within 2 weeks was more likely if the index event was a transient ischemic attack rather than a stroke. Access to early endarterectomy varied markedly between hospitals across the province and improved over time from 2003 to 2006. CONCLUSIONS: In this hospital-based cohort, the majority of patients undergoing carotid endarterectomy after a transient ischemic attack or stroke had surgery delayed well beyond the period of maximum effectiveness. To enhance secondary stroke prevention, greater efforts are needed to minimize delays to diagnosis and surgical treatment for patients with symptomatic carotid stenosis. PMID- 19542058 TI - Judgments of critical care nurses about risk for secondary brain injury. AB - BACKGROUND: Interdisciplinary care for patients with traumatic brain injury focuses on treating the primary brain injury and limiting further brain damage from secondary injury. Intensive care unit nurses have an integral role in preventing secondary brain injury; however, little is known about factors that influence nurses' judgments about risk for secondary brain injury. OBJECTIVE: To investigate which physiological and situational variables influence judgments of intensive care unit nurses about patients' risk for secondary brain injury, management solely with nursing interventions, and management by consulting another member of the health care team. METHODS: A multiple segment factorial survey design was used. Vignettes reflecting the complexity of real-life scenarios were randomly generated by using different values of each independent variable. Surveys containing the vignettes were sent to nurses at 2 level I trauma centers. Multiple regression was used to determine which variables influenced judgments about secondary brain injury. RESULTS: Judgments about risk for secondary brain injury were influenced by a patient's oxygen saturation, intracranial pressure, cerebral perfusion pressure, mechanism of injury, and primary diagnosis, as well as by nursing shift. Judgments about interventions were influenced by a patient's oxygen saturation, intracranial pressure, and cerebral perfusion pressure and by nursing shift. The initial judgments made by nurses were the most significant variable predictive of follow-up judgments. CONCLUSIONS: Nurses need standardized, evidence-based content for management of secondary brain injury in critically ill patients with traumatic brain injury. PMID- 19542059 TI - A survey of nurses' beliefs about the medical emergency team system in a canadian tertiary hospital. AB - BACKGROUND: Nurses are the primary activators of the medical emergency team (MET). Although the MET system can empower nurses to seek help in managing acutely ill patients, few data on nurses' beliefs about the system are available. OBJECTIVE: To evaluate nurses' beliefs and behaviors about the MET system. METHODS: Nurses from a large academic hospital in Canada were surveyed (2 demography-related questions and 17 Likert-scale questions). RESULTS: Of 614 nurses employed on units participating in the MET system, 293 (47.7%) were approached and 275 completed the survey (response rate, 93.9%). Most respondents (84.2%) believed that the MET could prevent cardiopulmonary arrest in acutely ill patients, and 94% believed that the MET allowed them to seek help for patients they were worried about. Most nurses (75.9%) would call the responsible physician before activating the MET. Fifteen percent indicated reluctance to activate the MET because of fear of criticism, but only 2.2% considered the MET overused. Most (81.3%) believed that the MET did not increase their workload, and 91.3% did not believe that the MET reduced their skills. Forty-eight percent of nurses indicated that they would activate the MET for a patient they were worried about, even if the patient had normal vital signs. CONCLUSION: Nurses value the MET system. Nurses believe that the MET can help them care for acutely ill patients and improve outcomes. However, barriers to MET activation exist, including a fear of criticism and an adherence to a more traditional model of first contacting the responsible physician before activating the MET. PMID- 19542061 TI - The descriptive epidemiology of congenital and acquired cryptorchidism in a UK infant cohort. AB - INTRODUCTION: Recent studies in other European countries suggest that the prevalence of congenital cryptorchidism continues to increase. This study aimed to explore the prevalence and natural history of congenital cryptorchidism in a UK centre. METHODS: Between October 2001 and July 2008, 784 male infants were born in the prospective Cambridge Baby Growth Study. 742 infants were examined by trained research nurses at birth; testicular position was assessed using standard techniques. Follow-up assessments were completed at ages 3, 12, 18 and 24 months in 615, 462, 393 and 326 infants, respectively. RESULTS: The prevalence of cryptorchidism at birth was 5.9% (95% CI 4.4% to 7.9%). Congenital cryptorchidism was associated with earlier gestational age (p<0.001), lower birth weight (p<0.001), birth length (p<0.001) and shorter penile length at birth (p<0.0001) compared with other infants, but normal size after age 3 months. The prevalence of cryptorchidism declined to 2.4% at 3 months, but unexpectedly rose again to 6.7% at 12 months as a result of new cases. The cumulative incidence of "acquired cryptorchidism" by age 24 months was 7.0% and these cases had shorter penile length during infancy than other infants (p = 0.003). CONCLUSIONS: The prevalence of congenital cryptorchidism was higher than earlier estimates in UK populations. Furthermore, this study for the first time describes acquired cryptorchidism or "ascending testis" as a common entity in male infants, which is possibly associated with reduced early postnatal androgen activity. PMID- 19542065 TI - Towards evidence based medicine for paediatricians. Is cranio-sacral therapy useful in the management of crying babies? PMID- 19542067 TI - Towards evidence based medicine for paediatricians. Role of antibiotic line locks in the treatment of infected central venous access devices. PMID- 19542068 TI - Towards evidence based medicine for paediatricians. How effective is varicella zoster immunoglobulin (VZIG) in preventing chickenpox in neonates following perinatal exposure? PMID- 19542069 TI - Adolescents with altitude: young people do well on Kilimanjaro. PMID- 19542070 TI - A regional database for autism spectrum disorders. PMID- 19542071 TI - Do we neglect infant safety during air travel? PMID- 19542074 TI - Functional outcomes after the Ross (pulmonary autograft) procedure assessed with magnetic resonance imaging and cardiopulmonary exercise testing. AB - OBJECTIVE: To assess autograft, homograft and ventricular function, as well as exercise capacity, in adult patients who have undergone the Ross procedure. SETTING: Single centre paediatric and adult congenital heart disease unit. Patients 45 subjects (24.6 years, range 16.9-52.2 years) who underwent the Ross procedure between 1994 and 2006 (8.1 years after the Ross operation, range 2.0 14.0 years). Interventions Cardiovascular magnetic resonance imaging, echocardiography and cardiopulmonary exercise testing. MAIN OUTCOME MEASURES: Autograft and homograft stenosis, and regurgitation. Autograft size. Biventricular function, scar volume and exercise capacity. RESULTS: Mean autograft regurgitation was 6.8%+/-8.3% (trivial regurgitation) and diameter was 40.0+/-7.0 mm. Mean homograft velocity was 2.4+/-0.6 m/s (mild-moderate stenosis) and regurgitation was 6.1%+/-8.3% (trivial regurgitation). Biventricular systolic function was normal (LV EF 63.1+/-6.4% and RV EF 60.1%+/-7.6%). In 38% of cases there was evidence of LV scar, mostly noted within the inter-ventricular septum. The mean exercise capacity achieved was 87%+/-22% of predicted. There was no correlation between exercise capacity and ventricular function or scar. CONCLUSION: This study demonstrates minor autograft and homograft dysfunction in the majority of patients after the Ross procedure, associated with good ventricular function and exercise capacity. In addition, minor scar was present in a third of patients with no functional consequences. PMID- 19542075 TI - Staphylococcus pasteuri bacteraemia in a patient with leukaemia. PMID- 19542076 TI - Hazard ratio funnel plots for survival comparisons. AB - BACKGROUND: Funnel plots are a form of control chart that give a snapshot of many institutions at a particular moment in time. This paper describes how funnel plots may be constructed for survival analyses based on hazard ratios obtained from a Cox regression model with adjustment for covariates and allowance for overdispersion. METHOD: Analysis of simulated and real survival data. RESULTS: It describes how centred hazard ratio estimates adjusted for covariates can be obtained from a Cox regression and gives details of the necessary programming in Stata. Allowance for overdispersion can be made by multiplying the standard errors by a factor based on either the model or the log-rank chi(2) statistics. Simulated results and a real example are presented. CONCLUSION: Funnel plots based on hazard ratios are easier to interpret than multiple Kaplan-Meier survival plots, and in contrast to funnel plots based on survival at, say, 5 years, are less open to accusations of bias and use more information. The interpretation of such plots may be enhanced by using standard meta-analysis methods. Hazard ratio comparisons may now be added to the repertoire of techniques used by Cancer Registries, Primary Care Trusts, and other commissioners of healthcare. PMID- 19542077 TI - A cross-national comparative perspective on racial inequities in health: the USA versus Canada. AB - BACKGROUND: Cross-national comparisons allow the examination of the malleability of associations between race and health. Racial inequities in chronic conditions, indicators of health status and behavioural risk factors between two similar advanced capitalist countries were compared. It was hypothesised that racial inequities will be mitigated in Canada compared with the USA. METHODS: Population based, cross-sectional data from the 2002-3 Joint Canada-USA Survey of Health (JCUSH) with 4953 adult respondents from the USA and 3455 from Canada. Models adjusted for age, sex, foreign birth, marital status, health insurance, education, income and home ownership. RESULTS: Compared with the USA, racial inequities in health were attenuated in Canada. In the USA, racial inequities in chronic diseases and fair or poor self-rated health were largely driven by inequities found among the native born. Strikingly, in Canada, however, there were few significant racial inequities and those occurred exclusively among the foreign born. Within strata of race and foreign birth, Canadians fared better, with both white people and non-white people reporting better health than their American counterparts. Foreign-born Canadians and Americans were more similar to each other in terms of health than native-born Canadians and Americans. Only among the native born did American white people and American non-white people have higher adjusted odds of hypertension, diabetes and obesity than Canadian white people and Canadian non-white people respectively. Self-rated health was worse for non-white Americans than non-white Canadians regardless of foreign birth. CONCLUSION: The influence of race on health is context dependent. There is no necessary link between race and a variety of health indicators. PMID- 19542078 TI - TP53 PIN3 and MDM2 SNP309 polymorphisms as genetic modifiers in the Li-Fraumeni syndrome: impact on age at first diagnosis. AB - BACKGROUND: Li-Fraumeni and Li-Fraumeni-like syndromes (LFS/LFL), characterised by the development of multiple early onset cancers with heterogeneous tumour patterns, are associated with germline TP53 mutations. Polymorphisms in the TP53 pathway (TP53 PEX4 at codon 72, rs1042522; MDM2 SNP309, rs2279744) have modifier effects on germline TP53 mutations that may account for the individual and familial diversity of tumour patterns. METHODS AND RESULTS: Four polymorphisms were analysed in a series of 135 Brazilian LFS/LFL cancer patients (32 TP53 mutation carriers and 103 wild-type subjects). We report for the first time that another polymorphism in the TP53 gene, TP53 PIN3 (rs17878362), has a strong modifier effect on germline TP53 mutations. This polymorphism, which consists of a 16 bp duplication in intron 3 (A1, non-duplicated allele; A2, duplicated allele), is associated with a difference of 19.0 years in the mean age at the first diagnosis in TP53 mutation carriers (n = 25, A1A1: 28.0 years; n = 7, A1A2: 47.0 years; p = 0.01). In addition, cancer occurrence before the age of 35 years is exclusively observed in A1A1 homozygotes. In this series, the effect of TP53 PEX4 and MDM2 SNP309 on age at diagnosis was similar to the one reported in other series and was smaller than the one of TP53 PIN3 (TP53 PIN3: difference of 19.0 years; TP53 PEX4: 8.3 years; MDM2 SNP309: 12.5 years). CONCLUSION: These results suggest that TP53 PIN3 is another polymorphism in the TP53 pathway that may have a modifier effect on germline TP53 mutations and may contribute to the phenotypic diversity of germline TP53 mutations associated with LFS/LFL patients. PMID- 19542080 TI - Addition of pathology and biomarker information significantly improves the performance of the Manchester scoring system for BRCA1 and BRCA2 testing. AB - BACKGROUND: Selection for genetic testing of BRCA1/BRCA2 is an important area of healthcare. Although testing costs for mutational analysis are falling, costs in North America remain in excess of US$3000 (UK price can be 690 pounds). Guidelines in most countries use a 10-20% threshold of detecting a mutation in BRCA1/2 combined within a family before mutational analysis is considered. A number of computer-based models have been developed. However, use of these models can be time consuming and difficult. The Manchester scoring system was developed in 2003 to simplify the selection process without losing accuracy. METHODS: In order to increase accuracy of prediction, breast pathology of the index case was incorporated into the Manchester scoring system based on 2156 samples from unrelated non-Jewish patients fully tested for BRCA1/2, and the scores were adapted accordingly. Results/ DISCUSSION: Data from breast pathology allowed adjustment of BRCA1 and combined BRCA1/2 scores alone. There was a lack of pathological homogeneity for BRCA2, therefore specific pathological correlates could not be identified. Upward adjustments in BRCA1 mutation prediction scores were made for grade 3 ductal cancers, oestrogen receptor (ER) and triple-negative tumours. Downward adjustments in the score were made for grade 1 tumours, lobular cancer, ductal carcinoma in situ and ER/HER2 positivity. Application of the updated scoring system led to four and nine more mutations in BRCA1 being identified at the 10% and 20% threshold, respectively. Furthermore, 65 and 58 fewer cases met the 10% and 20% threshold, respectively, for testing. Moreover, the adjusted score significantly improved the trade-off between sensitivity and specificity for BRCA1/2 prediction. PMID- 19542079 TI - Defective complex I assembly due to C20orf7 mutations as a new cause of Leigh syndrome. AB - BACKGROUND: Leigh syndrome is an early onset, progressive, neurodegenerative disorder with developmental and motor skills regression. Characteristic magnetic resonance imaging abnormalities consist of focal bilateral lesions in the basal ganglia and/or the brainstem. The main cause is a deficiency in oxidative phosphorylation due to mutations in an mtDNA or nuclear oxidative phosphorylation gene. METHODS AND RESULTS: A consanguineous Moroccan family with Leigh syndrome comprise 11 children, three of which are affected. Marker analysis revealed a homozygous region of 11.5 Mb on chromosome 20, containing 111 genes. Eight possible mitochondrial candidate genes were sequenced. Patients were homozygous for an unclassified variant (p.P193L) in the cardiolipin synthase gene (CRLS1). As this variant was present in 20% of a Moroccan control population and enzyme activity was only reduced to 50%, this could not explain the rare clinical phenotype in our family. Patients were also homozygous for an amino acid substitution (p.L159F) in C20orf7, a new complex I assembly factor. Parents were heterozygous and unaffected sibs heterozygous or homozygous wild type. The mutation affects the predicted S-adenosylmethionine (SAM) dependent methyltransferase domain of C20orf7, possibly involved in methylation of NDUFB3 during the assembly process. Blue native gel electrophoresis showed an altered complex I assembly with only 30-40% of mature complex I present in patients and 70-90% in carriers. CONCLUSIONS: A new cause of Leigh syndrome can be a defect in early complex I assembly due to C20orf7 mutations. PMID- 19542081 TI - A common variant in the adiponutrin gene influences liver enzyme values. AB - BACKGROUND: Two recent genome-wide association studies identified the liver expressed transmembrane protein adiponutrin to be associated with liver related phenotypes such as non-alcoholic fatty liver disease and liver function enzymes. These associations were not uniformly reported for various ethnicities. The aim of this study was to investigate a common non-synonymous variant within adiponutrin (rs738409, exon 3) with parameters of liver function in three independent West Eurasian study populations including a total of 4290 participants. METHODS: The study was performed in (1) the population based Bruneck Study (n=783), (2) the Salzburg Atherosclerosis Prevention Program in Subjects at High Individual Risk Study from Austria based on a healthy working population (n=1705), and the Utah Obesity Case-Control Study including a group of 1019 severely obese individuals (average body mass index 46.0 kg/m(2)) and 783 controls from the same geographical region of Utah. Liver enzymes measured were alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma glutamyl transferase (GGT). RESULTS: A strong recessive association of this polymorphism was found with age and gender adjusted ALT and AST concentrations: being homozygous for the minor allele resulted in a highly significant increase of ALT concentration of 3.53 U/l (p=1.86 x 10(-9)) and of AST concentration of 2.07 U/l (p=9.58 x 10(-6)), respectively. The associations were consistently found in all three study populations. CONCLUSION: The highly significant associations of this transversion polymorphism within the adiponutrin gene with increased ALT and AST concentrations support a role for adiponutrin as a susceptibility gene for hepatic dysfunction. PMID- 19542082 TI - Penetrance of marked cognitive impairment in older male carriers of the FMR1 gene premutation. AB - BACKGROUND: Male carriers of the FMR1 premutation are at risk of developing the fragile X-associated tremor/ataxia syndrome (FXTAS), a newly recognised and largely under-diagnosed late onset neurodegenerative disorder. Patients affected with FXTAS primarily present with cerebellar ataxia and intention tremor. Cognitive decline has also been associated with the premutation, but the lack of data on its penetrance is a growing concern for clinicians who provide genetic counselling. METHODS: The Mattis Dementia Rating Scale (MDRS) was administered in a double blind fashion to 74 men aged 50 years or more recruited from fragile X families (35 premutation carriers and 39 intrafamilial controls) regardless of their clinical manifestation. Based on previous publications, marked cognitive impairment was defined by a score or =65 yr) and young (< or =40 yr) surgical patients who received propofol until loss of consciousness (LOC) using stepwise increasing effect-site concentrations. LOC was defined as an observer assessment of alertness/sedation score <2, corresponding to the absence of response to mild prodding or shaking. RESULTS: We analysed 35 elderly [average age, 78 yr (range, 67-96)] and 34 young [35 (19-40)] patients. At LOC, all indices were significantly higher in elderly compared with young patients: BIS(LOC), median 70 (range, 58-91) vs 58 (40-70); SE(LOC), 71 (31-88) vs 55.5 (23-79); and RE(LOC), 79 (35-96) vs 59 (25-80) (P<0.001 for all comparisons). With all three monitors, only a minority of elderly patients lost consciousness within a 40-60 index range: two (5.7%) with BIS and RE each, and seven (20%) with SE. In young patients, the respective numbers were 20 (58.8%) for BIS, 13 (38.2%) for SE, and nine (26.5%) for RE. CONCLUSIONS: In adults undergoing propofol induction, BIS, SE, and RE indices at LOC are significantly affected by age. PMID- 19542104 TI - Prevention of propofol-induced pain in children: combination of alfentanil and lidocaine vs alfentanil or lidocaine alone. AB - BACKGROUND: Pain from a propofol injection is a common side-effect in paediatric patients. This prospective, randomized, double-blind study evaluated the efficacy of a combined pretreatment of alfentanil with lidocaine on the incidence and severity of propofol injection pain in children. METHODS: After obtaining parental consent, 120 paediatric patients were allocated randomly into one of the three groups (n=40, in each). The patients in the alfentanil group received alfentanil 15 microg kg(-1) 90 s before the propofol injection. The patients in the lidocaine group received propofol 3 mg kg(-1) premixed with lidocaine 0.1% over a 15 s period. The patients in the combination group received both alfentanil and lidocaine. RESULTS: The incidence of propofol injection pain (severity 2 or more) in the combination group (2.6%) was significantly lower than that in the alfentanil and lidocaine groups (30% and 38.5%, respectively) (P=0.001 and <0.001, respectively). No patient in the combination group complained of moderate or severe pain from propofol injection. CONCLUSIONS: Our study demonstrated that the combination treatment of two different analgesic modalities, alfentanil and lidocaine, could prevent the moderate and severe pain on propofol injection, and reduce the incidence of mild pain compared with each drug alone. PMID- 19542105 TI - Diabetic muscle infarction. PMID- 19542106 TI - Management of the effects of exposure to tear gas. PMID- 19542107 TI - Assessing candiduria in a critically ill patient. PMID- 19542109 TI - Pedobacter nyackensis sp. nov., Pedobacter alluvionis sp. nov. and Pedobacter borealis sp. nov., isolated from Montana flood-plain sediment and forest soil. AB - Three Gram-negative, rod-shaped, non-spore-forming eubacterial strains were isolated in western Montana, USA, and subjected to taxonomic studies. Strains NWG II14(T) and NWER-II11(T) were isolated from hyporheic sediments of a large alluvial flood plain, whereas strain G-1(T) was isolated from a conifer forest soil. On the basis of 16S rRNA gene sequence similarity, strains NWG-II14(T), NWER-II11(T) and G-1(T) were shown to belong to the family Sphingobacteriaceae and are most closely related to various species of the genus Pedobacter. The results of molecular, physiological and biochemical tests allowed genotypic and phenotypic differentiation of these three strains from 23 Pedobacter species with validly published names. The three isolates therefore represent novel species, for which the names Pedobacter nyackensis sp. nov. (type strain NWG-II14(T) =DSM 19625(T) =LMG 24260(T)), Pedobacter alluvionis sp. nov. (type strain NWER-II11(T) =DSM 19624(T) =LMG 24258(T)) and Pedobacter borealis sp. nov. (type strain G-1(T) =DSM 19626(T) =LMG 24259(T)) are proposed. PMID- 19542108 TI - Campylobacter cuniculorum sp. nov., from rabbits. AB - Eight strains of an unknown thermotolerant Campylobacter species were isolated from the caecal contents of rabbits (Oryctolagus cuniculus). All strains were initially identified as belonging to the genus Campylobacter by means of genus specific PCR, but none were identified using species-specific PCR for known thermophilic species. Cells were spiral shaped with bipolar unsheathed flagella, with no periplasmic fibres, and appeared coccoid after 10-12 days of incubation. Phylogenetic analyses based on 16S rRNA gene, rpoB and groEL sequences revealed that all strains formed a robust clade that was very distinct from recognized Campylobacter species. 16S rRNA gene sequence pairwise comparisons of strain 150B(T) with the type strains of other Campylobacter species revealed that the nearest phylogenetic neighbour was Campylobacter helveticus NCTC 12470(T), with 96.6 % similarity. The uniqueness of these rabbit isolates was confirmed by whole cell protein electrophoresis. Taken together, these data indicate that the strains belong to a novel Campylobacter species for which the name Campylobacter cuniculorum sp. nov. is proposed, with 150B(T) (=LMG 24588(T) =CCUG 56289(T)) as the type strain. PMID- 19542111 TI - Isolation of bacteria from the ileal mucosa of TNFdeltaARE mice and description of Enterorhabdus mucosicola gen. nov., sp. nov. AB - The diversity of bacteria associated with inflamed mucosa was investigated by culturing ileal samples from TNF(deltaARE) mice on a selective medium containing mucin. Among eight isolates, two strains (Mt1B3 and Mt1B8(T)) belonged to bacterial groups not yet cultured from the mouse intestine. Whereas strain Mt1B3 was identified as a member of the family Planococcaceae and is closely related to Sporosarcina species and Filibacter limicola DSM 13886(T), strain Mt1B8(T) was a novel bacterium. Based on phylogenetic analysis, strain Mt1B8(T) is a member of the family Coriobacteriaceae. The closest relatives with validly published names were Asaccharobacter celatus, Adlercreutzia equolifaciens (<96 % similarity) and Eggerthella species (<92 %). With respect to Asaccharobacter celatus and Eggerthella, the phylogenetic position of strain Mt1B8(T) was confirmed at the chemotaxonomic level by Fourier-transform infrared spectroscopic analysis. The major fatty acid of strain Mt1B8(T) is C(16 : 0) (23.9 %). Menaquinones were monomethylated. DNA-DNA relatedness between strain Mt1B8(T) and Asaccharobacter celatus DSM 18785(T) was 28 %. Strain Mt1B8(T) is a Gram-positive-staining rod that does not form spores and has a high DNA G+C content (64.2 mol%). Cells are aerotolerant but grow only under strictly anoxic conditions. They are sensitive to cefotaxime, clarithromycin, erythromycin, metronidazole, tetracycline, tobramycin and vancomycin. API and VITEK analysis showed the ability of strain Mt1B8(T) to convert a variety of amino acid derivatives. According to these findings, it is proposed to create a novel genus and species, Enterorhabdus mucosicola gen. nov., sp. nov., to accommodate strain Mt1B8(T). The type strain of Enterorhabdus mucosicola is Mt1B8(T) (=DSM 19490(T) =CCUG 54980(T)). PMID- 19542112 TI - Proposed minimal standards for describing new genera and species of the suborder Micrococcineae. AB - The Subcommittee on the Taxonomy of the Suborder Micrococcineae of the International Committee on Systematics of Prokaryotes has agreed on minimal standards for describing new genera and species of the suborder Micrococcineae. The minimal standards are intended to provide bacteriologists involved in the taxonomy of members of the suborder Micrococcineae with a set of essential requirements for the description of new taxa. In addition to sequence data comparisons of 16S rRNA genes or other appropriate conservative genes, phenotypic and genotypic criteria are compiled which are considered essential for the comprehensive characterization of new members of the suborder Micrococcineae. Additional features are recommended for the characterization and differentiation of genera and species with validly published names. PMID- 19542113 TI - Bacillus korlensis sp. nov., a moderately halotolerant bacterium isolated from a sand soil sample in China. AB - A Gram-positive-staining, rod-shaped, motile, spore-forming and moderately halotolerant bacterium, designated ZLC-26(T), was isolated from a sand soil sample collected from Xinjiang Province, China, and was characterized by using a polyphasic taxonomic approach. This isolate grew optimally at 30-37 degrees C and pH 7-8. It grew with 0-8% NaCl (optimum, 0-2 %). Comparative 16S rRNA gene sequence analysis showed that strain ZLC-26(T) was closely related to members of the genus Bacillus, exhibiting the highest 16S rRNA gene sequence similarities to Bacillus nealsonii DSM 15077(T) (97.1 %), B. shackletonii LMG 18435(T) (97.0 %), B. siralis 171544(T) (97.0 %), B. circulans IAM 12462(T) (96.7 %) and B. pocheonensis Gsoil 420(T) (96.7 %). Strain ZLC-26(T) contained MK-7 as the predominant menaquinone. The diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. The major cellular fatty acids were iso-C(15 : 0), C(16 : 1)omega11c and anteiso-C(15 : 0). The DNA G+C content was 38.2 mol%. These chemotaxonomic results supported the affiliation of strain ZLC 26(T) to the genus Bacillus. However, low DNA-DNA relatedness values and distinguishing phenotypic characteristics allowed genotypic and phenotypic differentiation of strain ZLC-26(T) from recognized Bacillus species. On the basis of the evidence presented, strain ZLC-26(T) is considered to represent a novel species of the genus Bacillus, for which the name Bacillus korlensis sp. nov. is proposed. The type strain is ZLC-26(T) (=CCTCC AB 207172(T)=NRRL B 51302(T)). PMID- 19542114 TI - Microbulbifer variabilis sp. nov. and Microbulbifer epialgicus sp. nov., isolated from Pacific marine algae, possess a rod-coccus cell cycle in association with the growth phase. AB - Phylogenetic and taxonomic characterization was performed for 14 strains of bacteria that produce anticancer antibiotics (pelagiomicins) (represented by strain Ni-2088(T)) and one strain that produces UV-absorbing substances (strain F 104(T)), isolated from marine algae and seagrass collected from coastal areas of tropical Pacific islands and a subtropical island of Japan. All 15 isolates were Gram-negative, strictly aerobic, non-motile and non-spore-forming. Sequence analysis of the 16S rRNA gene showed that the isolates occupied positions in the phylogenetic radiation of the genus Microbulbifer, with similarities of 93.6-97.6 %. The cells possessed a clearly discernible rod-coccus cell cycle in association with the growth phase; cells were rods during the growth phase and all converted to coccoid-ovoid cells when proliferation ceased. The coccoid-ovoid cells were optically denser than the rod cells and were viable for extended periods. They were considered to constitute a resting form. The type strains of described species of Microbulbifer were also found to possess identical rod-coccus cell cycles. The G+C content of the DNA was 48.1-49.7 mol%. The major respiratory quinone system was ubiquinone-8. The major fatty acids were C(18 : 1)omega7c and C(16 : 0), and the hydroxy acids comprised C(10 : 0) 3-OH, C(12 : 0) 3-OH and iso C(11 : 0) 3-OH. The polar lipids comprised phosphatidylethanolamine, phosphatidylglycerol and phosphatidylserine. The group of 14 pelagiomicin producing strains and strain F-104(T) each constituted a single genomic species. Based on phylogenetic affiliation, phenotypic characteristics and genomic distinctness, the isolates represent two novel species in the genus Microbulbifer, for which the names Microbulbifer variabilis sp. nov. (type strain Ni-2088(T) =MBIC01082(T) =ATCC 700307(T)) and Microbulbifer epialgicus sp. nov. (type strain F-104(T) =MBIC03330(T) =DSM 18651(T)) are proposed. PMID- 19542115 TI - Pontibacillus halophilus sp. nov., a moderately halophilic bacterium isolated from a sea urchin. AB - A Gram-positive-staining, moderately halophilic, strictly aerobic, catalase- and oxidase-positive, rod-shaped bacterium, designated strain JSM 076056(T), was isolated from a sea urchin collected from the South China Sea. Cells were motile by means of peritrichous flagella and formed ellipsoidal endospores lying in subterminal swollen sporangia. Strain JSM 076056(T) was able to grow at salinities of 2-25 % (w/v) total salts and at pH 6.0-10.0 and 15-40 degrees C; optimum growth was observed with 5-10 % (w/v) total salts and at pH 7.0-8.0 and 25-30 degrees C. It was incapable of growing with NaCl as the sole salt. meso Diaminopimelic acid was present in the cell-wall peptidoglycan. The major cellular fatty acids were iso-C(15 : 0), iso-C(16 : 0), iso-C(14 : 0,) anteiso C(15 : 0) and C(16 : 1)omega7c alcohol. The predominant respiratory quinone was MK-7 and the genomic DNA G+C content was 45.5 mol%. A phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JSM 076056(T) belonged to the family Bacillaceae and was related most closely to the type strains of the two recognized species of the genus Pontibacillus, namely Pontibacillus chungwhensis BH030062(T) (96.4 % sequence similarity) and Pontibacillus marinus BH030004(T) (96.2 %); these three strains formed a robust cluster in the phylogenetic tree. In combination, the phylogenetic, phenotypic and chemotaxonomic data indicate that strain JSM 076056(T) represents a novel species of the genus Pontibacillus, for which the name Pontibacillus halophilus sp. nov. is proposed. The type strain is JSM 076056(T) (=CCTCC AA 207029(T) =DSM 19796(T) =KCTC 13190(T)). PMID- 19542116 TI - Oceanibaculum indicum gen. nov., sp. nov., isolated from deep seawater of the Indian Ocean. AB - A taxonomic study was carried out on strain P24(T), which was isolated from a polycyclic aromatic hydrocarbon-degrading consortium, enriched from a deep seawater sample collected from the Indian Ocean. The isolate was Gram-negative, rod-shaped, motile by means of a polar flagellum, moderately halophilic and capable of reducing nitrate to nitrite. Growth was observed at salinities of 0-9 % and at temperatures of 10-42 degrees C. The strain was unable to degrade Tween 80 or gelatin. The dominant fatty acids were C(16 : 0) (15.2 % of the total), C(18 : 0) (10.3 %), C(18 : 1)omega7c (52.0 %), C(18 : 1) 2-OH (4.7 %) and C(19 : 0)omega8c cyclo (4.7 %). The G+C content of the chromosomal DNA was 64.8 mol%. 16S rRNA gene sequence comparisons showed that strain P24(T) was related most closely to Thalassobaculum litoreum CL-GR58(T) (92.7 % similarity); levels of similarity between strain P24(T) and type strains of recognized species in the family Rhodospirillaceae were all less than 90.8 %. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain P24(T) formed a distinct evolutionary lineage within the family Rhodospirillaceae. Strain P24(T) could be distinguished from phylogenetically related genera based on differences in several phenotypic properties. On the basis of the phenotypic and phylogenetic data presented, strain P24(T) is considered to represent a novel species of a new genus, for which the name Oceanibaculum indicum gen. nov., sp. nov. is proposed. The type strain is P24(T) (=CCTCC AB 208226(T)=LMG 24626(T)=MCCC 1A02083(T)). PMID- 19542117 TI - Differentiation of species of the family Acetobacteraceae by AFLP DNA fingerprinting: Gluconacetobacter kombuchae is a later heterotypic synonym of Gluconacetobacter hansenii. AB - Amplified fragment length polymorphism (AFLP) DNA fingerprinting was investigated as a tool for fast and accurate identification of acetic acid bacteria (AAB) to the species level. One hundred and thirty five reference strains and 15 additional strains, representing 50 recognized species of the family Acetobacteraceae, were subjected to AFLP analysis using the restriction enzyme combination ApaI/TaqI and the primer combination A03/T03. The reference strains had been previously subjected to either DNA-DNA hybridization or 16S-23S rRNA spacer region gene sequence analysis and were regarded as being accurately classified at the species level. The present study revealed that six of these strains should be reclassified, namely Gluconacetobacter europaeus LMG 1518 and Gluconacetobacter xylinus LMG 1510 as Gluconacetobacter xylinus and Gluconacetobacter europaeus, respectively; Gluconacetobacter kombuchae LMG 23726(T) as Gluconacetobacter hansenii; and Acetobacter orleanensis strains LMG 1545, LMG 1592 and LMG 1608 as Acetobacter cerevisiae. Cluster analysis of the AFLP DNA fingerprints of the reference strains revealed one cluster for each species, showing a linkage level below 50 % with other clusters, except for Acetobacter pasteurianus, Acetobacter indonesiensis and Acetobacter cerevisiae. These three species were separated into two, two, and three clusters, respectively. At present, confusion exists regarding the taxonomic status of Gluconacetobacter oboediens and Gluconacetobacter intermedius; the AFLP data from this study supported their classification as separate taxa. The 15 additional strains could all be identified at the species level. AFLP analysis further revealed that some species harboured genetically diverse strains, whereas other species consisted of strains showing similar banding patterns, indicating a more limited genetic diversity. It can be concluded that AFLP DNA fingerprinting is suitable for accurate identification and classification of a broad range of AAB, as well as for the determination of intraspecific genetic diversity. PMID- 19542118 TI - Planobacterium taklimakanense gen. nov., sp. nov., a member of the family Flavobacteriaceae that exhibits swimming motility, isolated from desert soil. AB - A Gram-negative and rod-shaped bacterial strain, X-65(T), isolated from desert soil of Xinjiang, China, was subjected to phenotypic and phylogenetic studies. On the basis of 16S rRNA gene sequence analyses, the isolate showed closest relationship to members of the genera Chryseobacterium (92.4-95.4 % similarity), Cloacibacterium (94.0 %), Sejongia (93.9 %) and Elizabethkingia (93.8 %) and to Riemerella columbina LMG 11607(T) (93.3 %). The strain differed from its phylogenetic neighbours by its swimming motility in liquid cultures and the presence of long appendages and large amounts of the straight-chain fatty acids 16 : 0 and 18 : 0. Growth was observed at 18-37 degrees C (optimum 30 degrees C), at pH 6-12 (optimum pH 7-8) and in the presence of 0-3 % NaCl (optimum 0 %). Flexirubin pigments were absent. The major isoprenoid quinone was MK-6. The DNA G+C content of X-65(T) was 41.5 mol%. The data presented in this study indicate that the isolate should be classified in a novel genus and species within the family Flavobacteriaceae, for which the name Planobacterium taklimakanense gen. nov., sp. nov. is proposed. The type strain of Planobacterium taklimakanense is X 65(T) (=CCTCC AB 208154(T) =NRRL B-51322(T)). PMID- 19542119 TI - Marinobacter lacisalsi sp. nov., a moderately halophilic bacterium isolated from the saline-wetland wildfowl reserve Fuente de Piedra in southern Spain. AB - A Gram-negative, non-spore-forming, motile, moderately halophilic, aerobic, rod shaped bacterium, designated strain FP2.5(T), was isolated from the inland hypersaline lake Fuente de Piedra, a saline-wetland wildfowl reserve located in the province of Malaga in southern Spain. Strain FP2.5(T) was subjected to a polyphasic taxonomic study. It produced colonies with a light-yellow pigment. Strain FP2.5(T) grew at salinities of 3-15 % (w/v) and at temperatures of 20-40 degrees C. The pH range for growth was 5-9. Strain FP2.5(T) was able to utilize various organic acids as sole carbon and energy source. Its major fatty acids were C(16 : 0), C(18 : 1)omega9c and C(16 : 1)omega9c. The DNA G+C content was 58.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain FP2.5(T) appeared to be a member of the genus Marinobacter and clustered closely with the type strains of Marinobacter segnicrescens, Marinobacter bryozoorum and Marinobacter gudaonensis (levels of 16S rRNA gene sequence similarity of 98.1, 97.4 and 97.2 %, respectively). However, DNA-DNA relatedness between the new isolate and the type strains of its closest related Marinobacter species was low; levels of DNA-DNA relatedness between strain FP2.5(T) and M. segnicrescens LMG 23928(T), M. bryozoorum DSM 15401(T) and M. gudaonensis DSM 18066(T) were 36.3, 32.1 and 24.9 %, respectively. On the basis of phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data, strain FP2.5(T) is considered to represent a novel species of the genus Marinobacter, for which the name Marinobacter lacisalsi sp. nov. is proposed. The type strain is FP2.5(T) (=CECT 7297(T)=LMG 24237(T)). PMID- 19542120 TI - Flavobacterium chungangense sp. nov., isolated from a freshwater lake. AB - A yellow-pigmented, rod-shaped, Gram-staining-negative, aerobic bacterial strain, designated CJ7(T), was isolated from a freshwater lake at Chung-Ang University in Anseong, South Korea. Results from 16S rRNA gene sequence analysis revealed that the isolate was phylogenetically affiliated with members of the genus Flavobacterium. Strain CJ7(T) showed sequence similarity values of 91.5-98.0 % to the type strains of recognized Flavobacterium species. The DNA-DNA relatedness between strain CJ7(T) and Flavobacterium hercynium DSM 18292(T), which showed the highest 16S rRNA gene sequence similarity, was 25.4 %. The predominant cellular fatty acids were iso-C(15 : 0) (18.3 %), summed feature 3 (comprising iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c) (16.1 %), iso-C(17 : 0) 3-OH (8.9 %), iso-C(15 : 0) 3-OH (7.2 %), iso-C(17 : 1)omega9c (6.1 %) and iso-C(15 : 1) (5.9 %). Flexirubin-type pigments were absent. The major isoprenoid quinone was menaquinone-6. The DNA G+C content was 34.5 mol%. The results obtained from this study, with a polyphasic taxonomic approach, suggested that strain CJ7(T) represents a novel species of the genus Flavobacterium for which the name Flavobacterium chungangense sp. nov. is proposed. The type strain is CJ7(T) (=KACC 13353(T)=JCM 15651(T)). PMID- 19542121 TI - Pseudozyma pruni sp. nov., a novel ustilaginomycetous anamorphic fungus from flowers in Taiwan. AB - Two ustilaginomycetous anamorphic strains were isolated from flowers in Taiwan. Phylogenetic analysis based on the combined rRNA gene sequence of internal transcribed spacer 1 (ITS1)-5.8S-ITS2 and large-subunit D1/D2 domains indicated that the closest recognized species was Pseudozyma fusiformata. The results of DNA-DNA hybridization and physiological characteristics showed that the two strains represent a novel species within the genus Pseudozyma. The name Pseudozyma pruni sp. nov. is proposed, with FIRDI 005(T) (=BCRC 34227(T) =CBS 10937(T)) as the type strain. PMID- 19542122 TI - Paenibacillus tundrae sp. nov. and Paenibacillus xylanexedens sp. nov., psychrotolerant, xylan-degrading bacteria from Alaskan tundra. AB - Eight psychrotolerant, xylan-degrading strains of bacteria that were catalase positive, oxidase-negative and able to reduce nitrate to nitrite were isolated from soil beneath moist non-acidic and acidic tundra in northern Alaska. The DNA G+C contents for the strains ranged from 46.4-50.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that each strain belonged to the genus Paenibacillus. The highest level of 16S rRNA gene similarity was found between the eight strains and Paenibacillus amylolyticus NRRL NRS-290(T) (98.9-99.1 %). However, despite relatively high 16S rRNA gene similarity, DNA-DNA hybridization, repetitive elements genotyping and phenotypic analysis revealed that at least two of the strains differed from P. amylolyticus NRRL NRS-290(T). DNA-DNA hybridization values between strain A10b(T) and P. amylolyticus NRRL NRS-290(T) (4.3 %), between strain B22a(T) and P. amylolyticus NRRL NRS-290(T) (48.8 %) and between strain A10b(T) and strain B22a(T) (11.0 %) were below those recommended by the ad hoc committee for those belonging to the same species. Significant phenotypic features that differentiate these novel strains from P. amylolyticus included their inability to utilize l-arabinose and ability to utilize glycogen as sole carbon sources. Unlike strains 1B4a and B22a(T), strains A6a and A10b(T) produced ethanol as an end product of glucose fermentation, utilized acetic acid and 2,3-butanediol and did not utilize d-gluconic acid. MK-7 was the major isoprenoid quinone and anteiso-C(15 : 0) was the most abundant fatty acid for strains A10b(T) and B22a(T). On the basis of these results, strains A10b(T) and B22a(T) are each considered to represent a novel species of the genus Paenibacillus, for which the names Paenibacillus tundrae sp. nov. and Paenibacillus xylanexedens sp. nov. are proposed, respectively. The type strain of Paenibacillus tundrae sp. nov. is A10b(T) (=NRRL B-51094(T)=DSM 21291(T)). The type strain of Paenibacillus xylanexedens sp. nov. is B22a(T) (=NRRL B 51090(T)=DSM 21292(T)). PMID- 19542123 TI - Clostridium sulfidigenes sp. nov., a mesophilic, proteolytic, thiosulfate- and sulfur-reducing bacterium isolated from pond sediment. AB - A novel strictly anaerobic, endospore-forming, rod-shaped, Gram-positive bacterium, designated strain SGB2(T), was isolated from a mixed culture from a pond sediment during screening for sulfate-reducing bacteria capable of degrading cyanophycin (CGP). In this study, the taxonomic characterization of this mesophilic, proteolytic Clostridium isolate and the role which it, and its phylogenetic relatives, may play in peptide degradation and in the sulfur cycle are reported. Strain SGB2(T) was a commensal strain, utilizing CGP degradation products produced by other micro-organisms. Cells were motile until sporulation, forming oval, terminal spores that swell the cells. It showed optimum growth at 34 degrees C, pH 6.6 and in the absence of NaCl. Strain SGB2(T) utilized proteinaceous compounds such as peptone, Casamino acids, gelatin and trypticase soy, in addition to several amino acids and pyruvate. Utilization of many of these compounds was enhanced in the presence of thiosulfate. The isolate was unable to use any of the carbohydrates or alcohols investigated or CGP as carbon and energy sources. Thiosulfate and elemental sulfur were used as terminal electron acceptors. Phylogenetic analysis revealed that strain SGB2(T) belongs to the low-G+C-containing Clostridiales group. It exhibited 99 % 16S rRNA gene sequence similarity to its closest relatives Clostridium thiosulfatireducens Lup21(T) and Clostridium subterminale DSM 6970(T). DNA-DNA hybridization values with these two strains were 39.4 and 42.1 %, respectively. Based on phenotypic, genotypic and phylogenetic characteristics, we conclude that the isolate represents a novel species of the genus Clostridium, Clostridium sulfidigenes sp. nov. The type strain is SGB2(T) (=DSM 18982(T) =ATCC BAA-1538(T)). PMID- 19542124 TI - Butyricimonas synergistica gen. nov., sp. nov. and Butyricimonas virosa sp. nov., butyric acid-producing bacteria in the family 'Porphyromonadaceae' isolated from rat faeces. AB - Two bacterial strains, designated MT01(T) and MT12(T), isolated from rat faeces were characterized by using a polyphasic taxonomic approach that included analysis of their phenotypic and biochemical features, cellular fatty acid profiles, menaquinone profiles and phylogeny based on 16S rRNA gene sequences. The 16S rRNA gene sequence analysis showed that these strains were members of the family 'Porphyromonadaceae'. The strains shared 94 % 16S rRNA gene sequence similarity with each other and were related to Odoribacter splanchnicus NCTC 10825(T) (86-87 % sequence similarity). The strains consisted of obligately anaerobic, non-pigmented, non-spore-forming, non-motile, Gram-negative rods. Growth of the strains was inhibited on medium containing 20 % bile. The two strains produced significant levels of butyric and isobutyric acids as end products from glucose. Although the major cellular fatty acid of these two strains and O. splanchnicus JCM 15291(T) was iso-C(15 : 0), strains MT01(T) and MT12(T) showed a higher level of iso-C(15 : 0) (66 and 74 %, respectively) than did O. splanchnicus JCM 15291(T) (48 %). In addition, the ratios of iso-C(15 : 0) to anteiso-C(15 : 0) in whole-cell methanolysates of the two isolates were very much higher than that of O. splanchnicus JCM 15291(T). The major menaquinone of the isolates was MK-10. This menaquinone composition was different from those of other genera of the family 'Porphyromonadaceae', such as Barnesiella (predominant menaquinones: MK-11 and MK-12), Odoribacter (MK-9), Paludibacter (MK-8), Parabacteroides (MK-9 and MK-10), Porphyromonas (MK-9 and MK-10) and Tannerella (MK-10 and MK-11). Menaquinone composition is therefore an important chemotaxonomic characteristic of these micro-organisms. Strains MT01(T) and MT12(T) have DNA G+C contents of 46 mol%. On the basis of these data, strains MT01(T) and MT12(T) represent two novel species of a novel genus, for which the names Butyricimonas synergistica gen. nov., sp. nov. and Butyricimonas virosa sp. nov., respectively, are proposed. The type strains of B. synergistica and B. virosa are MT01(T) (=JCM 15148(T) =CCUG 56610(T)) and MT12(T) (=JCM 15149(T)=CCUG 56611(T)), respectively. PMID- 19542125 TI - Massilia niabensis sp. nov. and Massilia niastensis sp. nov., isolated from air samples. AB - Two bacterial isolates, designated strains 5420S-26(T) and 5516S-1(T), were recovered from air samples collected in Suwon, Korea. Cells of both strains were aerobic, Gram-negative, motile rods. Phylogenetically, these strains were positioned within the radius of the genus Massilia. 16S rRNA gene sequence analysis showed that the strains shared 97.3 % sequence similarity and had sequence similarities of 94.9-98.1 % with respect to type strains of species belonging to the genus Massilia. In DNA-DNA hybridization tests, the two strains showed <39 % relatedness with respect to strains of closely related species of the genus Massilia and 27 % relatedness to each other. Both strains contained Q-8 as the predominant isoprenoid quinone and possessed summed feature 3 (comprising C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH) as the major fatty acid. Strain 5516S 1(T) was found to contain the fatty acid C(20 : 0) (in small amounts), a feature that served to distinguish it from both 5420S-26(T) and recognized members of the genus Massilia. The DNA G+C contents of 5420S-26(T) and 5516S-1(T) were 67.8 and 66.6 mol%, respectively. Phylogenetic, phenotypic and chemotaxonomic data accumulated in this study revealed that 5420S-26(T) and 5516S-1(T) represent novel species of the genus Massilia, for which the names Massilia niabensis sp. nov. (type strain 5420S-26(T) =KACC 12632(T) =DSM 21312(T)) and Massilia niastensis sp. nov. (type strain 5516S-1(T) =KACC 12599(T) =DSM 21313(T)) are proposed, respectively. PMID- 19542126 TI - Gracilibacillus saliphilus sp. nov., a moderately halophilic bacterium isolated from a salt lake. AB - A novel Gram-positive-staining, aerobic, spore-forming, rod-shaped bacterium, designated strain YIM 91119(T), was isolated from Ebinur Lake in Xinjiang Province, north-west China. Cells were motile, produced terminal endospores and grew at pH 6.0-8.0 (optimally at pH 7.0), 4-45 degrees C (optimally at 28-37 degrees C) and 1-22 % (w/v) NaCl, (optimally at 10-15 %, w/v). Comparative 16S rRNA gene sequence analysis showed that strain YIM 91119(T) belongs to the genus Gracilibacillus, exhibiting the highest sequence similarity with respect to the type strain of Gracilibacillus orientalis (97.8 %); the next most similar 16S rRNA gene sequences were those of the type strains of Gracilibacillus boraciitolerans (96.8 %), Gracilibacillus dipsosauri (96.5 %) and Gracilibacillus halotolerans (95.8 %). DNA-DNA hybridization with G. orientalis AS 1.4250(T) showed a relatedness of 55 %. The major fatty acids of strain YIM 91119(T) were anteiso-C(15 : 0), iso-C(15 : 0) and anteiso-C(17 : 0). The peptidoglycan type was A1gamma (directly cross-linked meso-diaminopimelic acid). The genomic DNA G+C content was 40.1 mol% and the predominant respiratory quinone was MK-7. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. On the basis of the evidence from this polyphasic study, strain YIM 91119(T) represents a novel species of the genus Gracilibacillus, for which the name Gracilibacillus saliphilus sp. nov. is proposed, with YIM 91119(T) (=DSM 19802(T) =CCTCC AA 208015(T)) as the type strain. PMID- 19542127 TI - Tenacibaculum crassostreae sp. nov., isolated from the Pacific oyster, Crassostrea gigas. AB - A rod-shaped, yellow-pigmented, aerobic, Gram-negative bacterium, designated strain JO-1(T), was isolated from an apparently healthy Pacific oyster, Crassostrea gigas, collected at Wan Island, Korea. It grew at 15-37 degrees C (optimum 30 degrees C) only in the presence of sea salts. Strain JO-1(T) hydrolysed casein, Tween 80 and starch. The major fatty acids were iso-C(15 : 0) (23.8 %), summed feature 3 (comprising C(16 : 1)omega7c and/or iso-C(15 : 0) 2 OH; 14.5 %) and iso-C(15 : 1) G (14.1 %). Analysis of the 16S rRNA gene sequence indicated that strain JO-1(T) was a member of the genus Tenacibaculum in the family Flavobacteriaceae, with sequence similarity of 94.6-97.8 % to the type strains of recognized members of the genus. The G+C content of the genomic DNA was 31.4 mol%. DNA-DNA relatedness levels between strain JO-1(T) and the five closest relatives, Tenacibaculum litoreum KCCM 42115(T), T. lutimaris KCTC 12302(T), T. aestuarii KCTC 12569(T), T. mesophilum DSM 13764(T) and T. adriaticum JCM 14633(T), were less than 28 %. Phylogenetic analyses and differences in physiological and biochemical characteristics suggested that strain JO-1(T) (=KCTC 22329(T) =JCM 15428(T)) should be classified as the type strain of a novel species within the genus Tenacibaculum, for which the name Tenacibaculum crassostreae sp. nov. is proposed. PMID- 19542129 TI - Desulfurococcus kamchatkensis sp. nov., a novel hyperthermophilic protein degrading archaeon isolated from a Kamchatka hot spring. AB - A novel obligately anaerobic, hyperthermophilic, organotrophic archaeon, designated strain 1221n(T), was isolated from a hot spring of Uzon Caldera (Kamchatka Peninsula, Russia). Cells of strain 1221n(T) were non-motile regular cocci, 0.6-1 mum in diameter. The temperature range for growth at pH 6.5 was 65 87 degrees C, with an optimum at 85 degrees C. The pH range for growth at 85 degrees C was 5.5-7.5, with an optimum at pH 6.5. Growth was not observed at or below 6 degrees C or at or above 90 degrees C, as well as at or below pH 5.0 and at or above pH 8.0. The isolate fermented a wide range of substrates including proteins: alpha-keratin, albumin and gelatin. Elemental sulfur was not essential for growth, but stimulated growth. Strain 1221n(T) synthesized 40 and 120 kDa proteinases localized on the cell envelope. The G+C content of the DNA was 44.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparison indicated that strain 1221n(T) was affiliated with the genus Desulfurococcus. The level of 16S rRNA gene sequence similarity with other Desulfurococcus species was 96.7 98.1 %, and Desulfurococcus amylolyticus was found to be the most closely related organism. Based on the data from the phylogenetic analysis and the physiological properties of the novel isolate, strain 1221n(T) should be classified as representing a novel species, for which the name Desulfurococcus kamchatkensis sp. nov. is proposed. The type strain is 1221n(T) (=DSM 18924(T)=VKM B-2413(T)). PMID- 19542128 TI - Streptosporangium canum sp. nov., isolated from soil. AB - An actinomycete strain, designated HBUM 170018(T), was isolated from soil from Hebei Province, China, and subjected to a polyphasic taxonomic analysis. This study included morphological and physiological investigations and analyses concerning cell chemistry, genomic DNA G+C content, DNA-DNA hybridization and phylogeny. The data obtained were consistent with the assignment of strain HBUM 170018(T) to the genus Streptosporangium and were confirmed by the results of 16S rRNA gene sequence analysis. Strain HBUM 170018(T) can be differentiated from all Streptosporangium species with validly published names with reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results. On the basis of 16S rRNA gene sequencing, DNA-DNA hybridization and phenotypic data, strain HBUM 170018(T) represents a novel species of the genus Streptosporangium, for which the name Streptosporangium canum sp. nov. is proposed. The type strain is HBUM 170018(T) (=7177(T) =DSM 45034(T) =CGMCC 4.2126(T)). PMID- 19542130 TI - Clostridium clariflavum sp. nov. and Clostridium caenicola sp. nov., moderately thermophilic, cellulose-/cellobiose-digesting bacteria isolated from methanogenic sludge. AB - Two novel anaerobic, moderately thermophilic and cellulose-/cellobiose-digesting bacteria, EBR45(T) and EBR596(T), were isolated from anaerobic sludge of a cellulose-degrading methanogenic bioreactor. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these strains belonged to cluster III within the low-G+C-content Gram-positive bacteria. The close relatives of EBR45(T) were Clostridium straminisolvens DSM 16021(T) (sequence identity, 94.6 %) and Clostridium thermocellum DSM 1237(T) (93.4 %). The closest relative of EBR596(T) was Clostridium stercorarium DSM 8532(T) (95.9 %). Both isolates were rod-shaped sporulators, growing optimally at 60 degrees C. EBR45(T) was Gram-staining reaction-variable and non-motile, formed bright-yellow colonies on solid media, and grew on a relatively narrow range of carbohydrates including cellulose and cellobiose. EBR596(T) was Gram-staining-reaction-negative and motile, formed glossy white colonies and grew on cellobiose and various carbohydrates except cellulose. Major fatty acid compositions were 16 : 0 iso, 16 : 0 and 16 : 0 dimethylacetal (strain EBR45(T)) and 15 : 0 iso, 16 : 0 iso, 15 : 0 anteiso and 17 : 0 anteiso (strain EBR596(T)). The DNA G+C contents were 36.9 mol% (EBR45(T)) and 51.1 mol% (EBR596(T)). Based on the phenotypic and phylogenetic data and genomic distinctiveness, strains EBR45(T) and EBR596(T) represent two novel species, for which the names Clostridium clariflavum sp. nov. (type strain EBR45(T) =DSM 19732(T) =NBRC 101661(T)) and Clostridium caenicola sp. nov. (type strain EBR596(T) =DSM 19027(T) =NBRC 102590(T)) are proposed. PMID- 19542131 TI - Parasutterella excrementihominis gen. nov., sp. nov., a member of the family Alcaligenaceae isolated from human faeces. AB - A novel, strictly anaerobic, non-spore-forming, Gram-negative coccobacillus (strain YIT 11859(T)) was isolated from human faeces. Biochemically, this strain was largely unreactive and was asaccharolytic. Growth of strain YIT 11859(T) in peptone-yeast extract broth produced no visible turbidity, and a trace amount of propionate was detected as an end product of metabolism. 16S rRNA gene sequence analysis showed that strain YIT 11859(T) was related most closely to the type strains of Sutterella species, with 90.8-88.0 % sequence similarity. Phylogenetic analysis of these and other related sequences confirmed that strain YIT 11859(T) was phylogenetically most closely associated with Sutterella species, but formed a separate cluster, indicating that strain YIT 11859(T) represents a novel member of the family Alcaligenaceae. Fatty acid analysis demonstrated the presence of a high concentration of C(18 : 1)omega9c (75 % of the total). The main respiratory quinones were menaquinone (MK-6) and methylated menaquinone (MMK-6). The G+C content of the DNA was 49.8 mol%. These results suggest that strain YIT 11859(T) represents a novel species of a new genus, for which the name Parasutterella excrementihominis gen. nov., sp. nov. is proposed. The type strain of Parasutterella excrementihominis is YIT 11859(T) (=DSM 21040(T) =JCM 15078(T)). PMID- 19542132 TI - Dyella soli sp. nov. and Dyella terrae sp. nov., isolated from soil. AB - Two novel strains isolated from soils, JS12-10(T) and JS14-6(T), were characterized using a polyphasic approach to determine their taxonomic positions. These isolates were found to be aerobic, Gram-negative, motile with one polar flagellum, non-spore-forming and rod-shaped. Phenotypic and fatty acid data supported the affiliation of JS12-10(T) and JS14-6(T) to the genus Dyella. However, chemotaxonomic data and DNA-DNA relatedness values allowed differentiation of these strains from other Dyella species with validly published names. Strains JS12-10(T) and JS14-6(T) showed the highest 16S rRNA gene sequence similarities with Dyella ginsengisoli Gsoil 3046(T) (98.4 %) and Dyella japonica XD53(T) (97.9 %), respectively, and the 16S rRNA gene sequence similarity between them was 97.1 %. DNA-DNA hybridization values between the novel isolates and strains of other recognized Dyella species were 29-38 %. Therefore, strains JS12 10(T) and JS14-6(T) represent two novel species of the genus Dyella, for which the names Dyella soli sp. nov. (type strain JS12-10(T) =KACC 12747(T) =JCM 15423(T)) and Dyella terrae sp. nov. (type strain JS14-6(T) =KACC 12748(T) =JCM 15424(T)) are proposed. PMID- 19542133 TI - Porphyromonas bennonis sp. nov., isolated from human clinical specimens. AB - During our investigation of the bacteriology of human wound infections and abscesses, a novel anaerobic, non-spore-forming, Gram-negative bacillus was frequently isolated. On the basis of morphological and biochemical criteria, the strains were tentatively identified as belonging to the family Bacteroidaceae, but they did not appear to correspond to any recognized species of this family. Comparative 16S rRNA gene sequencing showed that the 14 novel strains were genotypically homogeneous and confirmed their placement in the genus Porphyromonas. Sequence divergence values >10 % with respect to reference Porphyromonas species demonstrated that the strains isolated represent a novel species. On the basis of biochemical criteria and phylogenetic considerations, it is proposed that these strains isolated from human sources should be assigned to a novel species of the genus Porphyromonas, named Porphyromonas bennonis sp. nov., with WAL 1926C(T) (=ATCC BAA-1629(T) =CCUG 55979(T)) as the type strain. PMID- 19542134 TI - Jeotgalicoccus marinus sp. nov., a marine bacterium isolated from a sea urchin. AB - A novel non-sporulating, non-motile, catalase- and oxidase-positive, facultatively anaerobic, moderately halophilic, Gram-positive coccus, designated JSM 076033(T), was isolated from a sea urchin (Hemicentrotus pulcherrimus) collected from the South China Sea. Strain JSM 076033(T) was able to grow in the presence of 0.5-25.0 % (w/v) total salts and at pH 6.0-10.0 and 10-45 degrees C; optimum growth was observed with 5.0-10.0 % (w/v) total salts and at pH 7.0-8.0 and 25-30 degrees C. The major amino acid constituents of the cell wall were glycine, lysine and alanine. The major cellular fatty acids were anteiso-C(15 : 0), iso-C(15 : 0) and anteiso-C(17 : 0). The respiratory quinones were MK-7 (60.7 %) and MK-6 (39.3 %) and the polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unidentified phospholipid. The DNA G+C content was 40.3 mol%. A phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain JSM 076033(T) should be assigned to the genus Jeotgalicoccus. The sequence similarities between the novel isolate and the type strains of recognized Jeotgalicoccus species were in the range 95.2-97.2 %. The results of the phylogenetic analysis, combined with DNA-DNA relatedness data, phenotypic characteristics and chemotaxonomic information, support the view that strain JSM 076033(T) represents a novel species of the genus Jeotgalicoccus, for which the name Jeotgalicoccus marinus sp. nov. is proposed. The type strain is JSM 076033(T) (=CCTCC AA 207028(T) =DSM 19772(T) =KCTC 13189(T)). PMID- 19542135 TI - Algoriphagus aquatilis sp. nov., isolated from a freshwater lake. AB - A Gram-negative, non-spore-forming, non-motile bacterium, strain A8-7(T), was isolated from fresh water of a slightly alkaline lake, Longhu Lake, in Daqing, north-east China, and its taxonomic position was studied by using a polyphasic approach. Strain A8-7(T) was aerobic, heterotrophic and positive for catalase and oxidase. It grew at 20-37 degrees C (optimum 30 degrees C) and pH 5.5-10.5 (optimum pH 7.5) and in the presence of 0-3 % (w/v) NaCl. It formed pink pigmented, smooth and circular colonies, 1-2 mm in diameter, on R3A-V agar plates after incubation at 30 degrees C for 3 days. Cells of strain A8-7(T) were rods, 0.2-0.4 mum wide and 1.6-4.0 mum long. The major fatty acids (>10 %) were iso C(15 : 0) (40.3 %) and summed feature 3 (C(16 : 1)omega7c and/or iso-C(15 : 0) 2 OH; 12.1 %). The menaquinone was MK-7. The DNA G+C content was 43 mol% (T(m)). Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain A8 7(T) was phylogenetically related to members of the genus Algoriphagus, with sequence similarities of 92.6-95.2 %, the highest sequence similarity being to the sequence from Algoriphagus mannitolivorans IMSNU 14012(T). On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain A8-7(T) was considered to represent a novel species of the genus Algoriphagus, for which the name Algoriphagus aquatilis sp. nov. is proposed. The type strain is A8-7(T) (=CGMCC 1.7030(T) =NBRC 104237(T)). PMID- 19542137 TI - Adhaeribacter terreus sp. nov., isolated from forest soil. AB - A bacterial strain, designated DNG6(T), was isolated from forest soil of the Changbai mountains, Heilongjiang province, China. Cells of strain DNG6(T) were Gram-negative, 0.3-0.5 mum in diameter and 1.0-2.0 mum in length, strictly aerobic and produced large amounts of extracellular fibrillar material. Growth occurred at 16-33 degrees C (optimum, 28 degrees C), at pH 6.0-9.0 (optimum, pH 7.0-7.5) and in the presence of 0-1 % NaCl (optimum, 0 %). Strain DNG6(T) contained MK-7 as the major respiratory quinone and iso-C(15 : 0) (40.6 %) and summed feature 4 comprising iso-C(17 : 1) I and/or anteiso-C(17 : 1) B (26.5 %) as the major cellular fatty acids. The DNA G+C content was 48.1 mol% (T(m)). Phylogenetic analysis based on 16S rRNA gene sequences indicated that the closest relative of strain DNG6(T) was Adhaeribacter aquaticus MBRG1.5(T) with 93.9 % sequence similarity. Based on these results, it is concluded that strain DNG6(T) represents a novel species of the genus Adhaeribacter, for which the name Adhaeribacter terreus is proposed, with strain DNG6(T) (=CGMCC 1.6961(T)=NBRC 104235(T)) as the type strain. PMID- 19542136 TI - Vibrio azureus sp. nov., a luminous marine bacterium isolated from seawater. AB - Two luminous marine bacterial strains, LC2-005(T) and LC2-102, were isolated from seawater at Kuroshio Region and Sagami Bay in Japan, respectively. These bacteria were Gram-negative, oxidase-positive, catalase-positive, motile and rod-shaped. On the basis of 16S rRNA gene sequence analysis, strains LC2-005(T) and LC2-102 formed a cluster within the Vibrio harveyi species group. However, multilocus sequence analysis using five loci (pyrH, ftsZ, mreB, gyrB and gapA) and DNA-DNA hybridization experiments indicated that these strains were distinct from the currently known Vibrio species. Additionally, these strains differ from related Vibrio species in utilization of glucose, mannitol, inositol, sorbitol, rhamnose, sucrose, melibiose and arabinose, production of lysine decarboxylase, ornithine decarboxylase, tryptophan deaminase, esterase (C4), lipase (C4), chymotrypsin, acid phosphatase, alpha-glucosidase, beta-glucosidase and N-acetyl-beta glucosaminidase and the ability to reduce nitrate to nitrite. The major fatty acids were C(15 : 0) iso 2-OH and/or C(16 : 1)omega7c, C(16 : 0), C(18 : 1)omega7c and C(14 : 0). The DNA G+C contents of strains LC2-005(T) and LC2-102 were 45.2 and 45.5 mol%, respectively. On the basis of the polyphasic taxonomic evidence presented in this study, it can be concluded that strains LC2-005(T) and LC2-102 belong to the same genospecies and represent a novel species of the genus Vibrio, for which the name Vibrio azureus sp. nov. is proposed. The type strain is LC2-005(T) (=NBRC 104587(T) =KCTC 22352(T)). PMID- 19542138 TI - Promicromonospora flava sp. nov., isolated from sediment of the Baltic Sea. AB - A Gram-positive, non-spore-forming actinomycete, designated strain CC 0387(T), was isolated from a sediment sample from the Baltic Sea, Germany. Classification using a polyphasic approach and comparative 16S rRNA gene sequencing showed that strain CC 0387(T) belonged to the genus Promicromonospora and displayed more than 3 % 16S rRNA gene sequence divergence from all Promicromonospora species with validly published names. Strain CC 0387(T) did not produce aerial mycelium. Substrate mycelia were yellowish white to pale orange-yellow and fragmented into bacillary or coccoid elements. The cell wall contained lysine and alanine. Whole cell hydrolysates contained galactose, glucose, rhamnose and ribose. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, an unknown phospholipid and an unknown glycolipid. The predominant menaquinone was MK-9(H(4)) (86 %). The G+C content of the DNA was 71.87 mol%. Based on morphological, chemotaxonomic, phenotypic and genetic characteristics, strain CC 0387(T) (=CCTCC AA208024(T)=DSM 21481(T)) represents a novel species, for which the name Promicromonospora flava sp. nov. is proposed. PMID- 19542139 TI - Lachancea dasiensis sp. nov., an ascosporogenous yeast isolated from soil and leaves in Taiwan. AB - A novel yeast species, Lachancea dasiensis sp. nov., is proposed in this paper based on two strains isolated from leaves and one strain from soil in Taiwan. The strains produce one or two spherical ascospores in each ascus, preceded by conjugation of individual cells. Genus assignment and distinction of the species from other recognized species of Lachancea are based on morphological and physiological characteristics and on phylogenetic analysis of nucleotide sequences of the D1/D2 domains of the large subunit (LSU) rRNA gene. Sequence analysis of the D1/D2 domains of the LSU rRNA gene reveals that the phylogenetically closest relatives of L. dasiensis sp. nov. are Lachancea thermotolerans and Lachancea waltii. The novel species could be differentiated from the latter two species based on electrophoretic karyotypes. The type strain of Lachancea dasiensis sp. nov. is SC5L02(T) (=CBS 10888(T) =BCRC 23139(T)), which was isolated from a leaf of Angiopteris lygodiifolia Rosenst. in Dasi, Taoyuan, Taiwan. PMID- 19542140 TI - Microbacterium insulae sp. nov., isolated from soil. AB - A Gram-positive, non-motile, rod- or coccoid-shaped Microbacterium-like bacterium, designated strain DS-66(T), was isolated from soil of Dokdo, Korea, and its exact taxonomic position was investigated by using a polyphasic approach. Strain DS-66(T) grew optimally at 30 degrees C and pH 6.5-7.0 in the presence of 0.5-1.0 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain DS-66(T) belonged to the genus Microbacterium. Strain DS-66(T) had a peptidoglycan type based on B2beta with partial substitution of glutamic acid by 3-hydroxy glutamic acid (Glu/Hyg-Gly-d-Orn), and galactose, rhamnose and ribose as whole-cell sugars. The acyl type was glycolyl. Strain DS-66(T) contained MK-13, MK-12 and MK-14 as predominant menaquinones and anteiso-C(15 : 0), anteiso-C(17 : 0), iso-C(17 : 0) and iso-C(16 : 0) as major fatty acids. Major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid and an unidentified glycolipid. The DNA G+C content was 69.9 mol%. Phylogenetic distinctiveness, DNA-DNA relatedness data and differential phenotypic properties demonstrated that strain DS-66(T) is distinguishable from recognized Microbacterium species. On the basis of the data presented, strain DS-66(T) is considered to represent a novel species of the genus Microbacterium, for which the name Microbacterium insulae sp. nov. is proposed. The type strain is DS-66(T) (=KCTC 19247(T)=CCUG 54523(T)). PMID- 19542141 TI - Sphaerisporangium flaviroseum sp. nov. and Sphaerisporangium album sp. nov., isolated from forest soil in China. AB - Two Gram-positive, aerobic actinomycete strains, designated YIM 48771(T) and YIM 48782(T), were isolated from virgin forest soil samples collected in Hunan Province, China. 16S rRNA gene sequence similarities of the two novel isolates ranged from 96.3 to 97.6 % with species of the genus Sphaerisporangium with validly published names but, in the tree based on 16S rRNA gene sequences, the isolates formed distinct phyletic lines. The level of 16S rRNA gene sequence similarity between the two novel isolates was 97.1 %. DNA-DNA hybridization of strains YIM 48771(T) and YIM 48782(T) with recognized species of the genus Sphaerisporangium revealed that the level of DNA-DNA relatedness was below 70 %. The DNA G+C contents of strains YIM 48771(T) and YIM 48782(T) were 67.1 and 71 mol%, respectively. Chemotaxonomic data [major menaquinone, MK-9(H(4)); major polar lipids, diphosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylethanolamine and phosphoglycolipids; major fatty acids, iso-C(16 : 0) and 10-methyl C(17 : 0)] supported the affiliation of the two isolates with the genus Sphaerisporangium. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of the two isolates from recognized Sphaerisporangium species. Based on morphological, chemotaxonomical and phylogenetic data, strains YIM 48771(T) and YIM 48782(T) are considered to represent two novel species of the genus Sphaerisporangium, for which the names Sphaerisporangium flaviroseum sp. nov. (type strain, YIM 48771(T)=DSM 45170(T)=KCTC 19393(T)) and Sphaerisporangium album sp. nov. (type strain, YIM 48782(T)=DSM 45172(T)=CCTCC AA 208026(T)) are proposed. PMID- 19542144 TI - Sediminibacillus albus sp. nov., a moderately halophilic, Gram-positive bacterium isolated from a hypersaline lake, and emended description of the genus Sediminibacillus Carrasco et al. 2008. AB - A moderately halophilic, Gram-positive-staining, endospore-forming, rod-shaped and strictly aerobic bacterium, designated strain NHBX5(T), was isolated from Lake Nanhuobuxun in China. Strain NHBX5(T) was able to grow at NaCl concentrations of 0-22 % (w/v) (optimally at 7 %, w/v), at pH 6.0-9.0 (optimally at pH 7.5) and at temperatures of 10-45 degrees C (optimally at 37 degrees C). The cell-wall peptidoglycan of strain NHBX5(T) contained meso-diaminopimelic acid as the diagnostic diamino acid. The predominant isoprenoid quinone was MK-7. The major cellular fatty acids were anteiso-C(15 : 0) and anteiso-C(17 : 0). The polar lipids were phosphatidylglycerol, diphosphatidylglycerol and a glycolipid. The genomic DNA G+C content of strain NHBX5(T) was 44.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain NHBX5(T) was most closely related to Sediminibacillus halophilus EN8d(T) (98.6 % gene sequence similarity). The level of DNA-DNA relatedness between strain NHBX5(T) and S. halophilus CGMCC 1.6199(T) was 34.6 %. Based on the data presented, strain NHBX5(T) represents a novel species of the genus Sediminibacillus, for which the name Sediminibacillus albus sp. nov. is proposed. The type strain is NHBX5(T) (=DSM 19340(T) =CGMCC 1.6502(T)). In addition, an emended description of the genus Sediminibacillus is presented. PMID- 19542142 TI - Rhodovulum lacipunicei sp. nov., an obligate sulfide-demanding phototrophic alphaproteobacterium isolated from a purple pond in India. AB - An ovoid-rod-shaped, phototrophic, purple non-sulfur bacterium, designated strain JA322(T), was isolated in pure culture from a water sample collected from a saline pond with purple-coloured water, located near Satpada in Orissa, India. Strain JA322(T) was Gram-negative and non-motile and grew photoheterotrophically with a number of organic compounds serving as carbon sources or electron donors. Intracellular photosynthetic membranes were of the vesicular type. Bacteriochlorophyll a and carotenoids of the spheroidene series were present as photosynthetic pigments. Biotin and thiamine were required for growth of strain JA322(T). Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA322(T) was related most closely to the type strains of Rhodovulum euryhalinum (95.8 % similarity) and Rhodovulum strictum (94.8 %) and clustered with species of the genus Rhodovulum of the family Rhodobacteraceae. On the basis of phenotypic and molecular genetic evidence, it is suggested that strain JA322(T) represents a novel species of the genus Rhodovulum, for which the name Rhodovulum lacipunicei sp. nov. is proposed. The type strain is JA322(T) (=NBRC 104147(T)=KCTC 5700(T)). PMID- 19542146 TI - Vibrio breoganii sp. nov., a non-motile, alginolytic, marine bacterium within the Vibrio halioticoli clade. AB - Seven non-motile, facultatively anaerobic, alginolytic marine bacteria were isolated from the cultured clams Ruditapes philippinarum and Ruditapes decussatus. Phylogenetic analysis based on 16S rRNA gene sequences showed that these marine bacteria were closely related to the recently described species Vibrio comitans, Vibrio rarus and Vibrio inusitatus (>or=99.0 % sequence similarity). Phylogenetic analysis based on the housekeeping genes rpoA, recA and atpA grouped the isolates together and allocated them to the Vibrio halioticoli species group. Amplified fragment length polymorphism DNA fingerprinting also grouped them together and enabled them to be differentiated from recognized species of the V. halioticoli clade. DNA-DNA hybridizations showed that the isolates belonged to a novel species; phenotypic features such as the ability to grow at 4 degrees C and in the presence of 6 % NaCl also enabled them to be separated from other species. The DNA G+C content of RD 15.11(T) is 44.4 mol%. The genotypic and phenotypic data showed that the isolates represent a novel species in the V. halioticoli clade. The name Vibrio breoganii sp. nov. is proposed, with RD 15.11(T) (=CECT 7222(T) =LMG 23858(T)) as the type strain. PMID- 19542147 TI - Roseomonas frigidaquae sp. nov., isolated from a water-cooling system. AB - A non-motile, coccobacilli-shaped, pale-pink-pigmented bacterium, designated strain CW67(T), was isolated from a water-cooling system in Gwangyang, Republic of Korea. Cells were found to be Gram-negative, catalase-positive and oxidase positive, the major fatty acids were C(18 : 1)omega7c (43.6 %) and C(16 : 0) (15.8 %), the predominant respiratory lipoquinone was Q-10 and the DNA G+C content was 69.5 mol%. A phylogenetic tree based on 16S rRNA gene sequence comparisons showed that strain CW67(T) forms an evolutionary lineage within the radiation of the genus Roseomonas and that its closest relative is Roseomonas gilardii subsp. rosea MDA5605(T) (94.7 % sequence similarity). Evidence from this polyphasic study showed that strain CW67(T) could not be assigned to any recognized species. It therefore represents a novel species, for which the name Roseomonas frigidaquae sp. nov. is proposed, with CW67(T) (=KCTC 22211(T) =JCM 15073(T)) as the type strain. PMID- 19542148 TI - Increasing the coverage of a metapopulation consensus genome by iterative read mapping and assembly. AB - MOTIVATION: Most microbial species can not be cultured in the laboratory. Metagenomic sequencing may still yield a complete genome if the sequenced community is enriched and the sequencing coverage is high. However, the complexity in a natural population may cause the enrichment culture to contain multiple related strains. This diversity can confound existing strict assembly programs and lead to a fragmented assembly, which is unnecessary if we have a related reference genome available that can function as a scaffold. RESULTS: Here, we map short metagenomic sequencing reads from a population of strains to a related reference genome, and compose a genome that captures the consensus of the population's sequences. We show that by iteration of the mapping and assembly procedure, the coverage increases while the similarity with the reference genome decreases. This indicates that the assembly becomes less dependent on the reference genome and approaches the consensus genome of the multi-strain population. CONTACT: dutilh@cmbi.ru.nl SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19542149 TI - GR-Aligner: an algorithm for aligning pairwise genomic sequences containing rearrangement events. AB - MOTIVATION: Homologous genomic sequences between species usually contain different rearrangement events. Whether some specific patterns existed in the breakpoint regions that caused such events to occur is still unclear. To resolve this question, it is necessary to determine the location of breakpoints at the nucleotide level. The availability of sequences near breakpoints would further facilitate the related studies. We thus need a tool that can identify breakpoints and align the neighboring sequences. Although local alignment tools can detect rearrangement events, they only report a set of discontinuous alignments, where the detailed alignments in the breakpoint regions are usually missing. Global alignment tools are even less appropriate for these tasks since most of them are designed to align the conserved regions between sequences in a consistent order, i.e. they do not consider rearrangement events. RESULTS: We propose an effective and efficient pairwise sequence alignment algorithm, called GR-Aligner (Genomic Rearrangement Aligner), which can find breakpoints of rearrangement events by integrating the forward and reverse alignments of the breakpoint regions flanked by homologously rearranged sequences. In addition, GR-Aligner also provides an option to view the alignments of sequences extended to the breakpoints. These outputs provide materials for studying possible evolutionary mechanisms and biological functionalities of the rearrangement. PMID- 19542150 TI - FancyGene: dynamic visualization of gene structures and protein domain architectures on genomic loci. AB - SUMMARY: FancyGene is a fast and user-friendly web-based tool for producing images of one or more genes directly on the corresponding genomic locus. Starting from a variety of input formats, FancyGene rebuilds the basic components of a gene (UTRs, intron, exons). Once the initial representation is obtained, the user can superimpose additional features-such as protein domains and/or a variety of biological markers-in specific positions. FancyGene is extremely flexible allowing the user to change the resulting image dynamically, modifying colors and shapes and adding and/or removing objects. The output images are generated either in portable network graphics (PNG) or portable document format (PDF) formats and can be used for scientific presentations as well as for publications. The PDF format preserves editing capabilities, allowing picture modification using any vector graphics editor. PMID- 19542151 TI - VarScan: variant detection in massively parallel sequencing of individual and pooled samples. AB - SUMMARY: Massively parallel sequencing technologies hold incredible promise for the study of DNA sequence variation, particularly the identification of variants affecting human disease. The unprecedented throughput and relatively short read lengths of Roche/454, Illumina/Solexa, and other platforms have spurred development of a new generation of sequence alignment algorithms. Yet detection of sequence variants based on short read alignments remains challenging, and most currently available tools are limited to a single platform or aligner type. We present VarScan, an open source tool for variant detection that is compatible with several short read aligners. We demonstrate VarScan's ability to detect SNPs and indels with high sensitivity and specificity, in both Roche/454 sequencing of individuals and deep Illumina/Solexa sequencing of pooled samples. PMID- 19542152 TI - SHREC: a short-read error correction method. AB - MOTIVATION: Second-generation sequencing technologies produce a massive amount of short reads in a single experiment. However, sequencing errors can cause major problems when using this approach for de novo sequencing applications. Moreover, existing error correction methods have been designed and optimized for shotgun sequencing. Therefore, there is an urgent need for the design of fast and accurate computational methods and tools for error correction of large amounts of short read data. RESULTS: We present SHREC, a new algorithm for correcting errors in short-read data that uses a generalized suffix trie on the read data as the underlying data structure. Our results show that the method can identify erroneous reads with sensitivity and specificity of over 99% and 96% for simulated data with error rates of up to 3% as well as for real data. Furthermore, it achieves an error correction accuracy of over 80% for simulated data and over 88% for real data. These results are clearly superior to previously published approaches. SHREC is available as an efficient open-source Java implementation that allows processing of 10 million of short reads on a standard workstation. PMID- 19542153 TI - Copy number variation has little impact on bead-array-based measures of DNA methylation. AB - MOTIVATION: Integration of various genome-scale measures of molecular alterations is of great interest to researchers aiming to better define disease processes or identify novel targets with clinical utility. Particularly important in cancer are measures of gene copy number DNA methylation. However, copy number variation may bias the measurement of DNA methylation. To investigate possible bias, we analyzed integrated data obtained from 19 head and neck squamous cell carcinoma (HNSCC) tumors and 23 mesothelioma tumors. RESULTS: Statistical analysis of observational data produced results consistent with those anticipated from theoretical mathematical properties. Average beta value reported by Illumina GoldenGate (a bead-array platform) was significantly smaller than a similar measure constructed from the ratio of average dye intensities. Among CpGs that had only small variations in measured methylation across tumors (filtering out clearly biological methylation signatures), there were no systematic copy number effects on methylation for three and more than four copies; however, one copy led to small systematic negative effects, and no copies led to substantial significant negative effects. CONCLUSIONS: Since mathematical considerations suggest little bias in methylation assayed using bead-arrays, the consistency of observational data with anticipated properties suggests little bias. However, further analysis of systematic copy number effects across CpGs suggest that though there may be little bias when there are copy number gains, small biases may result when one allele is lost, and substantial biases when both alleles are lost. These results suggest that further integration of these measures can be useful for characterizing the biological relationships between these somatic events. PMID- 19542154 TI - Reconstructing signaling pathways from RNAi data using probabilistic Boolean threshold networks. AB - MOTIVATION: The reconstruction of signaling pathways from gene knockdown data is a novel research field enabled by developments in RNAi screening technology. However, while RNA interference is a powerful technique to identify genes related to a phenotype of interest, their placement in the corresponding pathways remains a challenging problem. Difficulties are aggravated if not all pathway components can be observed after each knockdown, but readouts are only available for a small subset. We are then facing the problem of reconstructing a network from incomplete data. RESULTS: We infer pathway topologies from gene knockdown data using Bayesian networks with probabilistic Boolean threshold functions. To deal with the problem of underdetermined network parameters, we employ a Bayesian learning approach, in which we can integrate arbitrary prior information on the network under consideration. Missing observations are integrated out. We compute the exact likelihood function for smaller networks, and use an approximation to evaluate the likelihood for larger networks. The posterior distribution is evaluated using mode hopping Markov chain Monte Carlo. Distributions over topologies and parameters can then be used to design additional experiments. We evaluate our approach on a small artificial dataset, and present inference results on RNAi data from the Jak/Stat pathway in a human hepatoma cell line. PMID- 19542157 TI - Age and Ageing. Editor's view. PMID- 19542156 TI - ISOLATE: a computational strategy for identifying the primary origin of cancers using high-throughput sequencing. AB - MOTIVATION: One of the most deadly cancer diagnoses is the carcinoma of unknown primary origin. Without the knowledge of the site of origin, treatment regimens are limited in their specificity and result in high mortality rates. Though supervised classification methods have been developed to predict the site of origin based on gene expression data, they require large numbers of previously classified tumors for training, in part because they do not account for sample heterogeneity, which limits their application to well-studied cancers. RESULTS: We present ISOLATE, a new statistical method that simultaneously predicts the primary site of origin of cancers and addresses sample heterogeneity, while taking advantage of new high-throughput sequencing technology that promises to bring higher accuracy and reproducibility to gene expression profiling experiments. ISOLATE makes predictions de novo, without having seen any training expression profiles of cancers with identified origin. Compared with previous methods, ISOLATE is able to predict the primary site of origin, de-convolve and remove the effect of sample heterogeneity and identify differentially expressed genes with higher accuracy, across both synthetic and clinical datasets. Methods such as ISOLATE are invaluable tools for clinicians faced with carcinomas of unknown primary origin. AVAILABILITY: ISOLATE is available for download at: http://morrislab.med.utoronto.ca/software CONTACT: gerald.quon@utoronto.ca; quaid.morris@utoronto.ca SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19542155 TI - Reconstruct modular phenotype-specific gene networks by knowledge-driven matrix factorization. AB - MOTIVATION: Reconstructing gene networks from microarray data has provided mechanistic information on cellular processes. A popular structure learning method, Bayesian network inference, has been used to determine network topology despite its shortcomings, i.e. the high-computational cost when analyzing a large number of genes and the inefficiency in exploiting prior knowledge, such as the co-regulation information of the genes. To address these limitations, we are introducing an alternative method, knowledge-driven matrix factorization (KMF) framework, to reconstruct phenotype-specific modular gene networks. RESULTS: Considering the reconstruction of gene network as a matrix factorization problem, we first use the gene expression data to estimate a correlation matrix, and then factorize the correlation matrix to recover the gene modules and the interactions between them. Prior knowledge from Gene Ontology is integrated into the matrix factorization. We applied this KMF algorithm to hepatocellular carcinoma (HepG2) cells treated with free fatty acids (FFAs). By comparing the module networks for the different conditions, we identified the specific modules that are involved in conferring the cytotoxic phenotype induced by palmitate. Further analysis of the gene modules of the different conditions suggested individual genes that play important roles in palmitate-induced cytotoxicity. In summary, KMF can efficiently integrate gene expression data with prior knowledge, thereby providing a powerful method of reconstructing phenotype-specific gene networks and valuable insights into the mechanisms that govern the phenotype. PMID- 19542158 TI - Abstracts of the British Geriatrics Society Autumn Scientific Meeting, 12-14 November 2008, Birmingham, UK. PMID- 19542175 TI - Disrupted gene pattern in patients with repeated in vitro fertilization (IVF) failure. AB - BACKGROUND: Repeated implantation failure (RIF) is a severe obstacle in human assisted reproduction treatment. METHODS: Aiming to identify global gene profile in RIF patients, gene-array analyses were performed on endometrial samples collected on day 21 of the cycle from fertile women (n = 12) and from RIF patients (n = 20). Validation of cyclin E2, Slug, dickkopf homolog 1 (DKK1), lymphoid enhancer-binding factor 1 (LEF1) and secreted frizzled-related protein 1 (SFRP1) was carried out by real-time PCR. RESULTS: Gene-array analysis revealed 313 genes exhibiting modified expression levels in RIF patients. Of these, 288 genes (92%) were down-regulated and only 25 genes (8%) were up-regulated. Classification of the down-regulated genes to biological pathways revealed cell cycle, Wnt signaling and cellular adhesion pathways. Real-time PCR validation of cyclin E2, SFRP1 and LEF1 showed significantly lower expression levels in RIF-IVF patients as compared with fertile women. In addition, two up-regulated genes, Slug and DKK1, were also validated. Interestingly, about 8% of the down-regulated genes were estrogen-dependent. Western blot of estrogen receptor alpha revealed low expression of this protein in the RIF group. CONCLUSIONS: The evaluation of the endometrium of RIF patients by gene array analysis demonstrates that the expression of various genes is altered, including those belonging to the cell cycle, Wnt signaling and cellular adhesion pathways. PMID- 19542176 TI - Inhibin alpha gene and susceptibility to premature ovarian failure: a data synthesis. AB - Candidate-gene association studies that examined the association between polymorphisms of the inhibin alpha gene (G769A, C16T and A124G) and premature ovarian failure (POF) have reported contradictory results. Thus, a meta-analysis of these studies was carried out. The random effects odds ratio (OR) with the corresponding 95% confidence interval and the heterogeneity among studies were estimated. Existence of potential bias and consistency of effect sizes across ethnicities were explored. Cumulative meta-analysis was also performed. The studies provided 1030/1660, 936/1398 and 938/1446 cases/controls for G769A, C16T and A124G polymorphisms, respectively. The meta-analysis showed significant heterogeneity among the studies (P(Q) = 0.01, I(2) = 74%) and lack of evidence that carriers of the G769A variant confer risk of POF: OR = 1.38 (0.48-3.94). Asian Indians (only two studies) produced significant association [OR = 8.10 (1.27-51.6)]. Regarding C16T and A124G polymorphisms, 16T and 124G alleles were not associated with POF: OR = 0.94 (0.76-1.16) and OR = 0.98 (0.86-1.11), respectively. The cumulative meta-analysis for G769A and C16T polymorphisms showed a trend in time towards to non-significance for both polymorphisms. Cumulative meta-analysis indicated that more evidence is needed to draw safer conclusions regarding the effect sizes. There was no differential magnitude of effect in large versus small studies. In conclusion, there is no evidence of association between the studied polymorphisms and POF. PMID- 19542177 TI - Mouse strain determines the outcome of wound healing after myocardial infarction. AB - AIMS: Our objective was to study the effect of the genetic background on the wound healing process after myocardial infarction (MI) in mice. METHODS AND RESULTS: MI was induced in five different mouse strains (BalbC, C57Bl6, FVB, 129S6, and Swiss). At 3, 14, and 28 days after MI, cardiac dimensions were monitored by echocardiography and histology, whereas cardiac function was determined by direct intraventricular pressure measurements (dP/dt). Furthermore, matrix metalloproteinases were measured by zymography, and mRNA expression by quantitative PCR. Infarct rupture, which typically occurred at 3-6 days post-MI, was most frequent in 129S6 mice (62%), followed by C57Bl6 (36%), FVB (29%), Swiss (23%), and BalbC (5%). The high incidence of infarct rupture in 129S6 mice was associated with high systolic blood pressure and increased influx of inflammatory cells. Cardiac dilatation was most marked in Swiss mice and least prominent in 129S6 mice. The degree of dilatation was associated with a reduced ejection fraction and decreased dP/dt values at 14 and 28 days post-MI. At day 14 and 28 post-MI, secondary thinning of the infarct area was marked in BalbC, FVB, and Swiss, but absent in C57Bl6 and 129S6 mice. In the latter two groups, this was paralleled by the highest number of myofibroblasts at day 14 post-MI. CONCLUSION: The outcome of infarct healing in mice strongly depends on genetic background. On the basis of our results, we suggest that for studies on infarct rupture, the 129S6 mouse is the background of choice, whereas BalbC and Swiss mice are the preferred models to study infarct thinning post-MI. PMID- 19542178 TI - Copper-induced regression of cardiomyocyte hypertrophy is associated with enhanced vascular endothelial growth factor receptor-1 signalling pathway. AB - AIMS: Vascular endothelial growth factor (VEGF) has been well documented to stimulate cell proliferation and differentiation; however, we have observed that copper (Cu)-induced regression of heart hypertrophy was VEGF-dependent. The present study was undertaken to test the hypothesis that Cu causes alterations in the distribution of VEGF receptors (VEGFRs) in hypertrophic cardiomyocytes so that it switches the signalling pathway from stimulation of cell growth to reversal of cell hypertrophy. METHODS AND RESULTS: Primary cultures of neonatal rat cardiomyocytes were exposed to phenylephrine (PE) at a final concentration of 100 microM in cultures for 48 h to induce cell hypertrophy. The hypertrophic cardiomyocytes were exposed to copper sulfate at a final concentration of 5 microM in cultures for 24 h with a concomitant presence of PE. Flow cytometry, gene silencing, and ELISA procedures were used to analyse the changes in VEGFRs and their relationship with regression of cardiomyocyte hypertrophy. Cu did not change the concentration of VEGF in culture media, but increased the ratio of VEGFR-1 to VEGFR-2 two-fold. Gene silencing of VEGFR-2, in the absence of Cu addition, reversed PE-induced cardiomyocyte hypertrophy, which was suppressed by an anti-VEGF antibody. Gene silencing of VEGFR-1 blocked Cu-induced regression of cell hypertrophy and decreased the activity of cGMP-dependent protein kinase-1 (PKG-1). A PKG-1 antagonist, Rp-8-pCPT-cGMPS, blocked both Cu- and VEGFR-2 gene silencing-induced regression of cardiomyocyte hypertrophy. CONCLUSION: Enhanced VEGFR-1 signalling is involved in Cu regression of cardiomyocyte hypertrophy, and the PKG-1 pathway is likely associated with VEGFR-1. PMID- 19542180 TI - A Synechocystis homolog of SipA protein, Ssl3451, enhances the activity of the histidine kinase Hik33. AB - In the cyanobacterium Synechocystis sp. PCC 6803, the histidine kinase Hik33 regulates the expression of several stress-inducible genes. Recently, a yeast two hybrid screen revealed a specific interaction between Hik33 and a small protein, Ssl3451. To investigate the function of Ssl3451, we developed an assay to monitor the autophosphorylation of Hik33 in vitro. Addition of Ssl3451 to the reaction mixture dramatically enhanced the autophosphorylation activity of Hik33. Pulse chase experiments revealed that Ssl3451 stimulated the autophosphorylation of Hik33 but did not affect its dephosphorylation. These findings indicated that Ssl3451 might be an activator of Hik33. When the amount of Hik33 was kept constant and the amount of Ssl3451 was increased in the reaction mixture, the extent of autophosphorylation of Hik33 reached a plateau when equimolar concentrations were present, suggesting that Ssl3451 enhances the activity of Hik33 by associating with it with a 1 : 1 stoichiometry. Disruption of the gene for Ssl3451 resulted in increased expression of the hliB gene, which is induced by Hik33 under standard growth conditions, but it did not affect the levels of the hliB mRNA at low temperature. Together, these results suggest that Ssl3451 might enhance the activity of Hik33 both in vitro and in vivo. PMID- 19542179 TI - d14, a strigolactone-insensitive mutant of rice, shows an accelerated outgrowth of tillers. AB - Recent studies using highly branched mutants of pea, Arabidopsis and rice have demonstrated that strigolactones, a group of terpenoid lactones, act as a new hormone class, or its biosynthetic precursors, in inhibiting shoot branching. Here, we provide evidence that DWARF14 (D14) inhibits rice tillering and may act as a new compo-nent of the strigolactone-dependent branching inhibition pathway. The d14 mutant exhibits increased shoot branch-ing with reduced plant height like the previously characterized strigolactone-deficient and -insensitive mutants d10 and d3, respectively. The d10-1 d14-1 double mutant is phenotypically indistinguishable from the d10-1 and d14-1 single mutants, consistent with the idea that D10 and D14 function in the same pathway. However, unlike with d10, the d14 branching phenotype could not be rescued by exogenous strigolactones. In addition, the d14 mutant contained a higher level of 2'-epi-5-deoxystrigol than the wild type. Positional cloning revealed that D14 encodes a protein of the alpha/beta-fold hydrolase superfamily, some members of which play a role in metabolism or signaling of plant hormones. We propose that D14 functions downstream of strigolactone synthesis, as a component of hormone signaling or as an enzyme that participates in the conversion of strigolactones to the bioactive form. PMID- 19542181 TI - Analysis of unintended events in hospitals: inter-rater reliability of constructing causal trees and classifying root causes. AB - BACKGROUND: Root cause analysis is a method to examine causes of unintended events. PRISMA (Prevention and Recovery Information System for Monitoring and Analysis: is a root cause analysis tool. With PRISMA, events are described in causal trees and root causes are subsequently classified with the Eindhoven Classification Model (ECM). It is important that root cause analysis tools are reliable, because they form the basis for patient safety interventions. OBJECTIVES: Determining the inter-rater reliability of descriptions, number and classifications of root causes. DESIGN: Totally, 300 unintended event reports were sampled from a database of 2028 events in 30 hospital units. The reports were previously analysed using PRISMA by experienced analysts and were re analysed to compare descriptions and number of root causes (n = 150) and to determine the inter-rater reliability of classifications (n = 150). MAIN OUTCOME MEASURES: Percentage agreement and Cohen's kappa (kappa). RESULTS: Agreement between descriptions of root causes was satisfactory: 54% agreement, 17% partial agreement and 29% no agreement. Inter-rater reliability of number of root causes was moderate (kappa = 0.46). Inter-rater reliability of classifying root causes with the ECM was substantial from highest category level (kappa = 0.71) to lowest subcategory level (kappa = 0.63). Most discrepancies occurred in classifying external causes. CONCLUSIONS: Results indicate that causal tree analysis with PRISMA is reliable. Analysts formulated similar root causes and agreed considerably on classifications, but showed variation in number of root causes. More training on disclosure of all relevant root causes is recommended as well as adjustment of the model by combining all external causes into one category. PMID- 19542182 TI - The Balanced Scorecard of acute settings: development process, definition of 20 strategic objectives and implementation. AB - CONTEXT: Strategy development and implementation in acute care settings is often restricted by competing challenges, the pace of policy reform and the existence of parallel hierarchies. OBJECTIVE: To describe a generic approach to strategy development, illustrate the use of the Balanced Scorecard as a tool to facilitate strategy implementation and demonstrate how to break down strategic goals into measurable elements. DESIGN: Multi-method approach using three different conceptual models: Health Promoting Hospitals Standards and Strategies, the European Foundation for Quality Management (EFQM) Model and the Balanced Scorecard. A bundle of qualitative and quantitative methods were used including in-depth interviews, standardized organization-wide surveys on organizational values, staff satisfaction and patient experience. SETTING: Three acute care hospitals in four different locations belonging to a German holding group. PARTICIPANTS: Chief executive officer, senior medical officers, working group leaders and hospital staff. INTERVENTION(S): Development and implementation of the Balanced Scorecard. MAIN OUTCOME MEASURE(S): Twenty strategic objectives with corresponding Balanced Scorecard measures. RESULTS: A stepped approach from strategy development to implementation is presented to identify key themes for strategy development, drafting a strategy map and developing strategic objectives and measures. CONCLUSIONS: The Balanced Scorecard, in combination with the EFQM model, is a useful tool to guide strategy development and implementation in health care organizations. As for other quality improvement and management tools not specifically developed for health care organizations, some adaptations are required to improve acceptability among professionals. The step-wise approach of strategy development and implementation presented here may support similar processes in comparable organizations. PMID- 19542183 TI - Antihypertension Induced by Tanshinone IIA Isolated from the Roots of Salvia miltiorrhiza. AB - Tanshinone IIA is one of the active principles in danshen (Salvia miltiorrhiza Bge) widely used in treatment of cardiovascular disorders. We investigated the effect of danshen or tanshinone IIA on blood pressure and its possible mechanisms. An i.p. injection of danshen at 10 mg kg(-1) significantly lowered systolic blood pressure (SBP) of spontaneously hypertensive rats (SHRs) but failed to modify the SBP in normotensive Wistar-Kyoto rats (WKY). Oral administration of tanshinone IIA also decreased SBP in SHR but not in WKY. Tanshinone IIA produced a concentration-dependent relaxation in isolated SHR aortic rings precontracted with phenylephrine (10 nmol l(-1)) or potassium chloride (KCl) (40 mmol l(-1)). The relaxing effect of tanshinone IIA on tonic contraction of phenylephrine in isolated aortic rings without endothelium remained produced. Glibenclamide at concentration sufficient to block adenosine triphosphatase (ATP)-sensitive potassium (K(+)) channel attenuated this tanshinone IIA-induced relaxation that was not influenced by other inhibitors. We further investigated the effect of tanshinone IIA on the changes of intracellular calcium concentration ([Ca(2+)](i)) in cultured aortic smooth muscle (A7r5) cells using fura-2 as indicator. Tanshinone IIA decreased [Ca(2+)](i) elicited by phenylephrine (10 nmol l(-1)) or KCl (40 mmol l(-1)) in a concentration-dependent manner; glibenclamide, but not other inhibitors for K(+) channel, abated this effect. Our results suggest that tanshinone IIA acts as an active principle of danshen showing vasodilation through ATP-sensitive K(+) channel to lower [Ca(2+)](i). PMID- 19542184 TI - Obesity and cancer: Mendelian randomization approach utilizing the FTO genotype. AB - BACKGROUND: Obesity is a risk factor for several cancers although appears to have an inverse association with cancers strongly related to tobacco. Studying obesity is difficult due to numerous biases and confounding. METHODS: To avoid these biases we used a Mendelian randomization approach incorporating an analysis of variants in the FTO gene that are strongly associated with BMI levels among 7000 subjects from a study of lung, kidney and upper-aerodigestive cancer. RESULTS: The FTO A allele which is linked with increased BMI was associated with a decreased risk of lung cancer (allelic odds ratio (OR) = 0.92, 95% confidence interval (CI) 0.84-1.00). It was also associated with a weak increased risk of kidney cancer, which was more apparent before the age of 50 (OR = 1.44, CI 1.09 1.90). CONCLUSION: Our results highlight the potential for genetic variation to act as an unconfounded marker of environmentally modifiable factors, and offer the potential to obtain estimates of the causal effect of obesity. However, far larger sample sizes than studied here will be required to undertake this with precision. PMID- 19542186 TI - A genome-wide library of CB4856/N2 introgression lines of Caenorhabditis elegans. AB - Recombinant inbred lines (RILs) derived from Caenorhabditis elegans wild-type N2 and CB4856 are increasingly being used for mapping genes underlying complex traits. To speed up mapping and gene discovery, introgression lines (ILs) offer a powerful tool for more efficient QTL identification. We constructed a library of 90 ILs, each carrying a single homozygous CB4856 genomic segment introgressed into the genetic background of N2. The ILs were genotyped by 123 single nucleotide polymorphism (SNP) markers. The proportion of the CB4856 segments in most lines does not exceed 3%, and together the introgressions cover 96% of the CB4856 genome. The value of the IL library was demonstrated by identifying novel loci underlying natural variation in two ageing-related traits, i.e. lifespan and pharyngeal pumping rate. Bin mapping of lifespan resulted in six QTLs, which all have a lifespan-shortening effect on the CB4856 allele. We found five QTLs for the decrease in pumping rate, of which four colocated with QTLs found for average lifespan. This suggests pleiotropic or closely linked QTL associated with lifespan and pumping rate. Overall, the presented IL library provides a versatile resource toward easier and efficient fine mapping and functional analyses of loci and genes underlying complex traits in C. elegans. PMID- 19542185 TI - Non-coding murine centromeric transcripts associate with and potentiate Aurora B kinase. AB - Non-coding RNAs are emerging as key players in many fundamental biological processes, including specification of higher-order chromatin structure. We examined the implication of RNA transcribed from mouse centromeric minor satellite repeats in the formation and function of centromere-associated complexes. Here we show that the levels of minor satellite RNA vary during cell cycle progression, peaking in G2/M phase, concomitant with accumulation of proteins of the chromosomal passenger complex near the centromere. Consistent with this, we describe that murine minor satellite RNA are components of CENP-A associated centromeric fractions and associate with proteins of the chromosomal passenger complex Aurora B and Survivin at the onset of mitosis. Interactions of endogenous Aurora B with CENP-A and Survivin are sensitive to RNaseA. Likewise, the kinase activity of Aurora B requires an RNA component. More importantly, Aurora B kinase activity can be potentiated by minor satellite RNA. In addition, decreased Aurora B activity after RNA depletion can be specifically rescued by restitution of these transcripts. Together, our data provide new functional evidence for minor satellite transcripts as key partners and regulators of the mitotic kinase Aurora B. PMID- 19542188 TI - Role of thyroid hormones in the developmental control of tissue glycogen in fetal sheep near term. AB - Developmental and glucocorticoid-induced changes in tissue glycogen content occur in the fetus near term coincident with an increase in plasma triiodothyronine (T(3)), although the role of thyroid hormones in mediating these changes is unknown. This study investigated glycogen content in the liver, heart and skeletal muscle of sheep fetuses after experimental manipulation of thyroid hormone concentration in utero by T(3) infusion and fetal thyroidectomy (TX). At 130 days of gestation (term 145 +/- 2 days), hepatic glycogen was greater, and muscle glycogen was lower, in the TX fetuses than in the intact fetuses. However, between 130 and 144 days of gestation the normal increment in hepatic glycogen, and decrement in cardiac glycogen, seen in intact fetuses was abolished when the prepartum rise in T(3), but not cortisol, was prevented by TX. At 144 days of gestation, hepatic glycogen was lower, and cardiac glycogen was higher, in the TX compared with intact fetuses. In intact fetuses at 130 days of gestation, 5 days of intravenous T(3) infusion (8-12 microg kg(-1) day(-1)) caused a small but significant increase in hepatic glycogen, although the concentration achieved was not as great as that observed in intact fetuses infused with cortisol (2-3 mg kg( 1) day(-1)) for 5 days. Infusion of T(3) reduced cardiac glycogen to the level observed in mature fetuses near term and immature fetuses infused with cortisol for 5 days. Glycogen content in fetal skeletal muscle increased between 100 and 115 days of gestation, but was unaffected by cortisol or T(3) infusion. Therefore, thyroid hormones are important in the developmental control of hepatic and cardiac glycogen content in the ovine fetus near term and may mediate, in part, the maturational effects of cortisol. PMID- 19542187 TI - A computational procedure to identify significant overlap of differentially expressed and genomic imbalanced regions in cancer datasets. AB - The integration of high-throughput genomic data represents an opportunity for deciphering the interplay between structural and functional organization of genomes and for discovering novel biomarkers. However, the development of integrative approaches to complement gene expression (GE) data with other types of gene information, such as copy number (CN) and chromosomal localization, still represents a computational challenge in the genomic arena. This work presents a computational procedure that directly integrates CN and GE profiles at genome wide level. When applied to DNA/RNA paired data, this approach leads to the identification of Significant Overlaps of Differentially Expressed and Genomic Imbalanced Regions (SODEGIR). This goal is accomplished in three steps. The first step extends to CN a method for detecting regional imbalances in GE. The second part provides the integration of CN and GE data and identifies chromosomal regions with concordantly altered genomic and transcriptional status in a tumor sample. The last step elevates the single-sample analysis to an entire dataset of tumor specimens. When applied to study chromosomal aberrations in a collection of astrocytoma and renal carcinoma samples, the procedure proved to be effective in identifying discrete chromosomal regions of coordinated CN alterations and changes in transcriptional levels. PMID- 19542189 TI - Feeding signals to the hungry mind. AB - Obesity, due to its associated co-morbidities, including type 2 diabetes and cardiovascular disease, is at the forefront of today's health care concerns. Our need for novel, multifaceted approaches to tackle the global increase of waistlines is urgent, and understanding the physiological processes underlying our vulnerability to weight gain is an important one of them. Evidence for considerable heritability of body weight indicates genetic influences in the susceptibility to our obesogenic environment. Here, we will focus on neurons in brain structures such as the hypothalamus, which sense the body's metabolic state and, through an intricate cascade of events, elicit an appropriate response. We will explore the use of genetically modified mouse models in the investigation of physiological functions of genes and pathways in neuronal regulation of metabolic balance. Use of these techniques allows us to make manipulations at the molecular level (e.g. in the neuronal metabolic sensing mechanism) and combine this with systems-level physiological analysis (e.g. body weight). Recent technological advances also enable the investigation of the contributions of genes to the co morbidities of obesity, such as obesity-induced hypertension. Reviewing examples of improvements as well as large gaps in our knowledge, this lecture aims to incite interest in whole body physiological research. PMID- 19542190 TI - Tea polyphenols decrease serum levels of prostate-specific antigen, hepatocyte growth factor, and vascular endothelial growth factor in prostate cancer patients and inhibit production of hepatocyte growth factor and vascular endothelial growth factor in vitro. AB - The purpose of this study was to determine the effects of short-term supplementation with the active compounds in green tea on serum biomarkers in patients with prostate cancer. Twenty-six men with positive prostate biopsies and scheduled for radical prostatectomy were given daily doses of Polyphenon E, which contained 800 mg of (-)-epigallocatechin-3-gallate (EGCG) and lesser amounts of ( )-epicatechin, (-)-epigallocatechin, and (-)-epicatechin-3-gallate (a total of 1.3 g of tea polyphenols), until time of radical prostatectomy. Serum was collected before initiation of the drug study and on the day of prostatectomy. Serum biomarkers hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), insulin-like growth factor (IGF)-I, IGF binding protein-3 (IGFBP 3), and prostate-specific antigen (PSA) were analyzed by ELISA. Toxicity was monitored primarily through liver function enzymes. Changes in serum components were analyzed statistically using the Wilcoxon signed rank test. Cancer associated fibroblasts were treated with EGCG, and HGF and VEGF protein and mRNA levels were measured. HGF, VEGF, PSA, IGF-I, IGFBP-3, and the IGF-I/IGFBP-3 ratio decreased significantly during the study. All of the liver function tests also decreased, five of them significantly: total protein, albumin, aspartate aminotransferase, alkaline phosphatase, and amylase. The decrease in HGF and VEGF was confirmed in prostate cancer-associated fibroblasts in vitro. Our results show a significant reduction in serum levels of PSA, HGF, and VEGF in men with prostate cancer after brief treatment with EGCG (Polyphenon E), with no elevation of liver enzymes. These findings support a potential role for Polyphenon E in the treatment or prevention of prostate cancer. PMID- 19542192 TI - The incidence rate of premature rupture of membranes and its influence on fetal neonatal health: a report from mainland China. AB - Premature rupture of membranes (PROM) is a common pregnancy complication and is associated with significant risks of fetal and neonatal morbidity and mortality. However, there is a lack of data concerning PROM in China. We investigated the incidence rate and analysed the influence of PROM on fetal and neonatal health. This used data from five specialist hospitals and one general hospital in mainland China. The total number of deliveries and the number of those complicated by PROM were recorded between January 2003 and December 2007. The time from initiation of PROM until delivery (latent period), the volume of amniotic fluid at delivery, fetal conditions and neonatal clinical conditions were recorded. The results suggest that the incidence of PROM was 19.53% and it could influence various aspects of the health of fetuses and neonates, including platelet parameters, erythrocyte parameters, neonatal jaundice and myocardial injury. PMID- 19542191 TI - Lower risk of cervical intraepithelial neoplasia in women with high plasma folate and sufficient vitamin B12 in the post-folic acid fortification era. AB - The purpose of this study was to determine the influence of plasma folate and vitamin B12 concentrations on cervical cancer risk in the U.S. after the folic acid fortification era. The study included 376 premenopausal women of childbearing age who tested positive for infections with high-risk (HR) human papillomaviruses (HPVs) and were diagnosed with cervical intraepithelial neoplasia (CIN) grade 2 or higher (CIN 2+, cases) or 19.8 ng/mL) who also had sufficient plasma vitamin B12 (>or=200.6 pg/mL) had 70% lower odds of being diagnosed with CIN 2+ (P = 0.04) when compared with women with plasma folate of 6 years) patients with severe peritonitis lasting for almost 1 month, nine patients after long-term PD (>6 years) without peritonitis and three normal subjects were included in the present study. Expressions of decorin, versican, hyaluronan, MMP-2, alfa smooth muscle actin (alphaSMA) and CD68 for macrophages in these specimens were examined by immunohistochemical staining. RESULTS: Although expression of decorin was detected in normal subjects, it was markedly decreased with long-term PD treatment. In long-term PD patients, the expression of versican was observed in their fibrotic-thickened peritoneum. Versican was present in fibrous regions, elastic lamina of the peritoneum, vascular walls and perivascular regions. Hyaluronan was observed in the whole thickened peritoneum, but its distribution differed in part from that of versican. MMP-2 was mainly observed around the blood vessels. Alfa SMA-positive cells, namely 'myofibroblasts' and CD68-positive cells, i.e. macrophages, were observed in the fibrotic-thickened peritoneum of long-term PD patients. Expressions of MMP-2, hyaluronan, SMA and CD68 in the peritoneum were marked in long-term PD patients' samples, which were strongly immunostained by versican, and were especially high in peritonitis patients. CONCLUSIONS: It appears that alterations in PGs, including marked induction of versican with peritonitis and disappearance of decorin, are involved in peritoneal remodelling in PD patients. Versican expression was closely related to the appearance of myofibroblasts and macrophages. These observations suggest that the alteration in PG components following PD therapy and severe inflammation contribute to fibrous thickening of the peritoneum. PMID- 19542197 TI - Independent responses to ultraviolet radiation and herbivore attack in broccoli. AB - The plant responses to ultraviolet-B radiation (UV-B) and to insect herbivory are believed to be partially similar. In this study, responses to these factors were investigated in the crop species broccoli (Brassica oleracea L. convar. botrytis, Brassicaceae). Plants were first grown under three UV-B regimes (80%, 23%, and 4% transmittance of ambient UV-B) in greenhouses covered with either innovative materials (high and medium transmittance) or conventional glass (low transmittance). Half of the plants then remained under these conditions, but the other half were transferred to the field with ambient light and herbivore access for up to 3 d. The plant responses to distinct environmental conditions were examined by analysing the morphological and chemical parameters of plants kept inside and plants exposed in the field. Furthermore, suitability of field-exposed plants to naturally occurring insects was investigated in relation to UV-B pretreatment. High levels of UV-B radiation led to increased flavonoid concentrations, but to a lower biomass accumulation in broccoli. These patterns remained after outdoor exposure. However, UV-induced changes of plant traits did not alter attractiveness to herbivorous insects: thrips, whiteflies, and aphids attacked plants independently of UV-B pretreatment. A 3-fold increase of indolyl glucosinolate concentrations occurred in above-ground tissue of all the plants, most likely due to massive herbivore attack after 3 d of field exposure. The results show that plants respond with high specificity to different abiotic and biotic impacts, demonstrating the separate perception and processing of stress factors. PMID- 19542198 TI - Transition to adult services among behaviorally infected adolescents with HIV--a qualitative study. AB - OBJECTIVES: The present study aimed to describe the experiences of youth with behaviorally acquired HIV who transitioned to adult care, to identify difficulties encountered, and to explore areas for improvement. METHODS: Semi structured interviews were conducted with 10 young adults ranging from 24 to 29 years old. Themes were derived from coding participant interviews. RESULTS: Participants experienced adolescent care providers as an important source of support, felt anxiety about transition, provided recommendations for improving the process, and described significant changes associated with adult HIV care. CONCLUSIONS: Findings support the development of a clear and structured transition process to address patients' fears and worries through early communication, planning, and coordination for adult healthcare, highlighting the need for future research in this area. PMID- 19542199 TI - Behavioral outcome in children with a history of neonatal encephalopathy following perinatal asphyxia. AB - OBJECTIVE: To examine the effects of mild and moderate neonatal encephalopathy (NE) on behavioral functioning, and prevalence of psychiatric diagnoses at 9-10 years. METHODS: The Child Behavior Checklist (CBCL), Teacher's Report Form (TRF), Diagnostic Interview Schedule for Children IV (DISC-IV), and the Children's Social Behavior Questionnaire (CSBQ) were used to assess behavioral outcome of 34 children with mild NE, 47 children with moderate NE, and 53 typically developing controls. RESULTS: Both children with mild and moderate NE showed more problematic behaviors than controls, which are related to a diversity of behavioral domains: elevated rates of social problems, anxiety and depression, attention regulation problems, and thought problems. No group differences were found in percentages of children with a DISC-IV (DSM-IV) classification. CONCLUSIONS: NE has a mildly negative effect on behavioral functioning, but does not lead to elevated levels or specific patterns of developmental psychopathology. PMID- 19542200 TI - Insulin storage and glucose homeostasis in mice null for the granule zinc transporter ZnT8 and studies of the type 2 diabetes-associated variants. AB - OBJECTIVE: Zinc ions are essential for the formation of hexameric insulin and hormone crystallization. A nonsynonymous single nucleotide polymorphism rs13266634 in the SLC30A8 gene, encoding the secretory granule zinc transporter ZnT8, is associated with type 2 diabetes. We describe the effects of deleting the ZnT8 gene in mice and explore the action of the at-risk allele. RESEARCH DESIGN AND METHODS: Slc30a8 null mice were generated and backcrossed at least twice onto a C57BL/6J background. Glucose and insulin tolerance were measured by intraperitoneal injection or euglycemic clamp, respectively. Insulin secretion, electrophysiology, imaging, and the generation of adenoviruses encoding the low- (W325) or elevated- (R325) risk ZnT8 alleles were undertaken using standard protocols. RESULTS: ZnT8(-/-) mice displayed age-, sex-, and diet-dependent abnormalities in glucose tolerance, insulin secretion, and body weight. Islets isolated from null mice had reduced granule zinc content and showed age-dependent changes in granule morphology, with markedly fewer dense cores but more rod-like crystals. Glucose-stimulated insulin secretion, granule fusion, and insulin crystal dissolution, assessed by total internal reflection fluorescence microscopy, were unchanged or enhanced in ZnT8(-/-) islets. Insulin processing was normal. Molecular modeling revealed that residue-325 was located at the interface between ZnT8 monomers. Correspondingly, the R325 variant displayed lower apparent Zn(2+) transport activity than W325 ZnT8 by fluorescence-based assay. CONCLUSIONS: ZnT8 is required for normal insulin crystallization and insulin release in vivo but not, remarkably, in vitro. Defects in the former processes in carriers of the R allele may increase type 2 diabetes risks. PMID- 19542201 TI - Tissue-specific remodeling of the mitochondrial proteome in type 1 diabetic akita mice. AB - OBJECTIVE: To elucidate the molecular basis for mitochondrial dysfunction, which has been implicated in the pathogenesis of diabetes complications. RESEARCH DESIGN AND METHODS: Mitochondrial matrix and membrane fractions were generated from liver, brain, heart, and kidney of wild-type and type 1 diabetic Akita mice. Comparative proteomics was performed using label-free proteome expression analysis. Mitochondrial state 3 respirations and ATP synthesis were measured, and mitochondrial morphology was evaluated by electron microscopy. Expression of genes that regulate mitochondrial biogenesis, substrate utilization, and oxidative phosphorylation (OXPHOS) were determined. RESULTS: In diabetic mice, fatty acid oxidation (FAO) proteins were less abundant in liver mitochondria, whereas FAO protein content was induced in mitochondria from all other tissues. Kidney mitochondria showed coordinate induction of tricarboxylic acid (TCA) cycle enzymes, whereas TCA cycle proteins were repressed in cardiac mitochondria. Levels of OXPHOS subunits were coordinately increased in liver mitochondria, whereas mitochondria of other tissues were unaffected. Mitochondrial respiration, ATP synthesis, and morphology were unaffected in liver and kidney mitochondria. In contrast, state 3 respirations, ATP synthesis, and mitochondrial cristae density were decreased in cardiac mitochondria and were accompanied by coordinate repression of OXPHOS and peroxisome proliferator-activated receptor (PPAR)-gamma coactivator (PGC)-1alpha transcripts. CONCLUSIONS: Type 1 diabetes causes tissue specific remodeling of the mitochondrial proteome. Preservation of mitochondrial function in kidney, brain, and liver, versus mitochondrial dysfunction in the heart, supports a central role for mitochondrial dysfunction in diabetic cardiomyopathy. PMID- 19542204 TI - Glutathione--a review on its role and significance in Parkinson's disease. AB - Parkinson's disease (PD) is the second most common neurodegenerative disease, affecting over a million people in the United States alone, and is characterized by rigidity, bradykinesia, resting tremor, and postural instability. Its main neuropathological feature is the loss of dopaminergic neurons of the substantia nigra pars compacta. However, the pathogenesis of this loss is not understood fully. One of the earliest biochemical changes seen in PD is a reduction in the levels of total glutathione, a key cellular antioxidant. Traditionally, it has been thought that this decrease in GSH levels is the consequence of increased oxidative stress, a process heavily implicated in PD pathogenesis. However, emerging evidence suggests that GSH depletion may itself play an active role in PD pathogenesis. This review aims to explore the contribution of GSH depletion to PD pathogenesis. PMID- 19542202 TI - SH2B1 enhances insulin sensitivity by both stimulating the insulin receptor and inhibiting tyrosine dephosphorylation of insulin receptor substrate proteins. AB - OBJECTIVE: SH2B1 is a SH2 domain-containing adaptor protein expressed in both the central nervous system and peripheral tissues. Neuronal SH2B1 controls body weight; however, the functions of peripheral SH2B1 remain unknown. Here, we studied peripheral SH2B1 regulation of insulin sensitivity and glucose metabolism. RESEARCH DESIGN AND METHODS: We generated TgKO mice expressing SH2B1 in the brain but not peripheral tissues. Various metabolic parameters and insulin signaling were examined in TgKO mice fed a high-fat diet (HFD). The effect of SH2B1 on the insulin receptor catalytic activity and insulin receptor substrate (IRS)-1/IRS-2 dephosphorylation was examined using in vitro kinase assays and in vitro dephosphorylation assays, respectively. SH2B1 was coexpressed with PTP1B, and insulin receptor-mediated phosphorylation of IRS-1 was examined. RESULTS: Deletion of peripheral SH2B1 markedly exacerbated HFD-induced hyperglycemia, hyperinsulinemia, and glucose intolerance in TgKO mice. Insulin signaling was dramatically impaired in muscle, liver, and adipose tissue in TgKO mice. Deletion of SH2B1 impaired insulin signaling in primary hepatocytes, whereas SH2B1 overexpression stimulated insulin receptor autophosphorylation and tyrosine phosphorylation of IRSs. Purified SH2B1 stimulated insulin receptor catalytic activity in vitro. The SH2 domain of SH2B1 was both required and sufficient to promote insulin receptor activation. Insulin stimulated the binding of SH2B1 to IRS-1 or IRS-2. This physical interaction inhibited tyrosine dephosphorylation of IRS-1 or IRS-2 and increased the ability of IRS proteins to activate the phosphatidylinositol 3-kinase pathway. CONCLUSIONS: SH2B1 is an endogenous insulin sensitizer. It directly binds to insulin receptors, IRS-1 and IRS-2, and enhances insulin sensitivity by promoting insulin receptor catalytic activity and by inhibiting tyrosine dephosphorylation of IRS proteins. PMID- 19542205 TI - Large gene family expansions and adaptive evolution for odorant and gustatory receptors in the pea aphid, Acyrthosiphon pisum. AB - Gaining insight into the mechanisms of chemoreception in aphids is of primary importance for both integrative studies on the evolution of host plant specialization and applied research in pest control management because aphids rely on their sense of smell and taste to locate and assess their host plants. We made use of the recent genome sequence of the pea aphid, Acyrthosiphon pisum, to address the molecular characterization and evolution of key molecular components of chemoreception: the odorant (Or) and gustatory (Gr) receptor genes. We identified 79 Or and 77 Gr genes in the pea aphid genome and showed that most of them are aphid-specific genes that have undergone recent and rapid expansion in the genome. By addressing selection within sets of paralogous Or and Gr expansions, for the first time in an insect species, we show that the most recently duplicated loci have evolved under positive selection, which might be related to the high degree of ecological specialization of this species. Although more functional studies are still needed for insect chemoreceptors, we provide evidence that Grs and Ors have different sets of positively selected sites, suggesting the possibility that these two gene families might have different binding pockets and bind structurally distinct classes of ligand. The pea aphid is the most basal insect species with a completely sequenced genome to date. The identification of chemoreceptor genes in this species is a key step toward further exploring insect comparative genetics, the genomics of ecological specialization and speciation, and new pest control strategies. PMID- 19542206 TI - Potential role of alpha-synuclein and metallothionein in lead-induced inclusion body formation. AB - Lead (Pb) produces aggresome-like inclusion bodies (IBs) in target cells as a toxic response. Our prior work shows metallothionein (MT) is required for this process. We used MT-I/II double knockout (MT-null) and parental wild-type (WT) cell lines to further explore the formation process of Pb-induced IBs. Unlike WT cells, MT-null cells did not form IBs after Pb exposure. Western blot of cytosol showed soluble MT protein in WT cells was lost during Pb exposure as IBs formed. Transfection of MT-I into MT-null cells allowed IBs formation after Pb exposure. Considering Pb-induced IBs may be like disease-related aggresomes, which often contain alpha-synuclein (Scna), we investigated Scna expression in cells capable (WT) and incapable (MT-null) of producing IBs after Pb exposure. Scna protein showed poor basal expression in MT-null cells. Pb exposure increased Scna expression only in WT cells. MT transfection increased Scna transcript to WT levels. In WT or MT-transfected MT-null cells, Pb-induced Scna expression rapidly increased and then decreased over 48 h as Pb-induced IBs were formed. A direct interaction between Scna and MT was confirmed ex vivo by antibody pulldown assay where the proteins coprecipitated with an antibody to MT. Pb exposure caused increased colocalization of MT and Scna proteins with time only in WT cells. In WT mice after chronic Pb exposure Scna was localized in renal cells containing forming IBs, whereas MT-null mice did not form IBs. Thus, Scna could be component of Pb-induced IBs and, with MT, may play a role in IBs formation. PMID- 19542207 TI - Sustained effects of interleukin-1 receptor antagonist treatment in type 2 diabetes. AB - OBJECTIVE: Interleukin (IL)-1 impairs insulin secretion and induces beta-cell apoptosis. Pancreatic beta-cell IL-1 expression is increased and interleukin-1 receptor antagonist (IL-1Ra) expression reduced in patients with type 2 diabetes. Treatment with recombinant IL-1Ra improves glycemia and beta-cell function and reduces inflammatory markers in patients with type 2 diabetes. Here we investigated the durability of these responses. RESEARCH DESIGN AND METHODS: Among 70 ambulatory patients who had type 2 diabetes, A1C >7.5%, and BMI >27 kg/m(2) and were randomly assigned to receive 13 weeks of anakinra, a recombinant human IL-1Ra, or placebo, 67 completed treatment and were included in this double blind 39-week follow-up study. Primary outcome was change in beta-cell function after anakinra withdrawal. Analysis was done by intention to treat. RESULTS: Thirty-nine weeks after anakinra withdrawal, the proinsulin-to-insulin (PI/I) ratio but not stimulated C-peptide remained improved (by -0.07 [95% CI -0.14 to 0.02], P = 0.011) compared with values in placebo-treated patients. Interestingly, a subgroup characterized by genetically determined low baseline IL 1Ra serum levels maintained the improved stimulated C-peptide obtained by 13 weeks of IL-1Ra treatment. Reductions in C-reactive protein (-3.2 mg/l [-6.2 to 1.1], P = 0.014) and in IL-6 (-1.4 ng/l [-2.6 to -0.3], P = 0.036) were maintained until the end of study. CONCLUSIONS: IL-1 blockade with anakinra induces improvement of the PI/I ratio and markers of systemic inflammation lasting 39 weeks after treatment withdrawal. PMID- 19542208 TI - Short sleep duration is associated with a blood pressure nondipping pattern in type 1 diabetes: the DIAPASOM study. AB - OBJECTIVE: To assess whether nocturnal blood pressure dipping status in type 1 diabetes is correlated with specific sleep characteristics and differences in nocturnal glycemic profiles. RESEARCH DESIGN AND METHODS: Twenty type 1 diabetic adult patients underwent sleep studies with simultaneous 24-h ambulatory blood pressure monitoring and continuous nocturnal glucose monitoring. RESULTS: Altogether, 55% of patients exhibited blunted blood pressure dipping. They did not differ from the dipper group in age, BMI, or systolic (SBP) and diastolic (DBP) blood pressure. Total sleep period (TSP) was higher in the dipper group (497 +/- 30 vs. 407 +/- 44 min for dippers and nondippers, respectively, P < 0.001). TSP was correlated with SBP and DBP day-night differences (r = 0.44 and 0.49, respectively). Periods of nocturnal hypoglycemia (i.e., % of TSP with glycemia <70 mg/dl) were longer in the dipper group (8.1 +/- 10.7 vs. 0.1 +/- 0.4% for dippers and nondippers, respectively, P = 0.02). CONCLUSIONS: Dipping status in type 1 diabetes was associated with longer sleep duration and with hypoglycemia unawareness. PMID- 19542209 TI - Cystic fibrosis-related diabetes: current trends in prevalence, incidence, and mortality. AB - OBJECTIVE: Cystic fibrosis (CF)-related diabetes (CFRD) diagnosis and management have considerably changed since diabetes was first shown to be associated with a poor prognosis in subjects with CF. Current trends in CFRD prevalence, incidence, and mortality were determined from a comprehensive clinical database. RESEARCH DESIGN AND METHODS: Data were reviewed from 872 CF patients followed at the University of Minnesota during three consecutive intervals: 1992-1997, 1998-2002, and 2003-2008. RESULTS: CFRD is currently present in 2% of children, 19% of adolescents, and 40-50% of adults. Incidence and prevalence are higher in female subjects aged 30-39 years; otherwise, there are no sex differences. In younger individuals, CFRD without fasting hyperglycemia predominates, but fasting hyperglycemia prevalence rises with age. CFRD mortality has significantly decreased over time. From 1992-1997 to 2003-2008, mortality rate in female subjects dropped by >50% from 6.9 to 3.2 deaths per 100 patient-years and in male subjects from 6.5 to 3.8 deaths per 100 patient-years. There is no longer a sex difference in mortality. Diabetes was previously diagnosed as a perimorbid event in nearly 20% of patients, but of 61 patients diagnosed with diabetes during 2003 2008, only 2 died. Lung function but not nutritional status is still worse in CF patients with diabetes compared with those without diabetes. Nutritional status and pulmonary status are similar between patients without fasting hyperglycemia and those with fasting hyperglycemia. CONCLUSIONS: Previously noted sex differences in mortality have disappeared, and the gap in mortality between CF patients with and without diabetes has considerably narrowed. We believe that early diagnosis and aggressive treatment have played a major role in improving survival in these patients. PMID- 19542210 TI - Impact of a natural disaster on diabetes: exacerbation of disparities and long term consequences. AB - OBJECTIVE: To examine the impact of Hurricane Katrina on the health of individuals with diabetes. RESEARCH DESIGN AND METHODS: This was an observational study in 1,795 adults with an A1C measurement 6 months before and 6-16 months after Hurricane Katrina in three health care systems: private (Tulane University Hospital and Clinic [TUHC]), state (Medical Center of Louisiana at New Orleans [MCLNO]), and Veterans Affairs (VA). Glycemic control (A1C), blood pressure, and lipids before the hurricane were compared with the patients' first measurement thereafter. The CORE Diabetes Model was used to project life expectancy and health economic impact. RESULTS: Mean predisaster A1C levels differed between MCLNO and VA patients (mean 7.7 vs. 7.3%, P < 0.001) and increased significantly among MCLNO patients to 8.3% (P < 0.001) but not among VA and TUHC patients. Mean systolic blood pressure increased in all three systems (130-137.6 mmHg for TUHC and 130.7-143.7 for VA, P < 0.001; 132-136 for MCLNO, P = 0.008). Mean LDL cholesterol increased in the VA (97.1-104.3 mg/dl) and TUHC patients (103.4 115.5; P < 0.001). Hurricane Katrina increased modeled direct, indirect, and total health care costs and also reduced life expectancy as well as quality adjusted life expectancy, with the economic impact being quite substantial because of the large population size affected. We estimate a lifetime cost of USD $504 million for the adult population affected, with the largest economic impact seen among MCLNO patients. CONCLUSIONS: A major disaster had a significant effect on diabetes management and exacerbated existing disparities. These effects may have a lasting impact on both health and economic implications. PMID- 19542211 TI - Elevated serum uric acid concentrations independently predict cardiovascular mortality in type 2 diabetic patients. AB - OBJECTIVE: There is limited information on whether increased serum uric acid levels are independently associated with cardiovascular mortality in type 2 diabetes. We assessed the predictive role of serum uric acid levels on all-cause and cardiovascular mortality in a large cohort of type 2 diabetic individuals. RESEARCH DESIGN AND METHODS: The cohort included 2,726 type 2 diabetic outpatients, who were followed for a mean period of 4.7 years. The independent association of serum uric acid levels with all-cause and cardiovascular mortality was assessed by Cox proportional hazards models and adjusted for conventional risk factors and several potential confounders. RESULTS: During follow-up, 329 (12.1%) patients died, 44.1% (n = 145) of whom from cardiovascular causes. In univariate analysis, higher serum uric acid levels were significantly associated with increased risk of all-cause (hazard ratio 19 [95% CI 1.12-1.27], P < 0.001) and cardiovascular (1.25 [1.16-1.34], P < 0.001) mortality. After adjustment for age, sex, BMI, smoking, hypertension, dyslipidemia, diabetes duration, A1C, medication use (allopurinol or hypoglycemic, antihypertensive, lipid-lowering, and antiplatelet drugs), estimated glomerular filtration rate, and albuminuria, the association of serum uric acid with cardiovascular mortality remained statistically significant (1.27 [1.01-1.61], P = 0.046), whereas the association of serum uric acid with all-cause mortality did not. CONCLUSIONS: Higher serum uric acid levels are associated with increased risk of cardiovascular mortality in type 2 diabetic patients, independent of several potential confounders, including renal function measures. PMID- 19542212 TI - The epidemiology of Takayasu arteritis in the UK. AB - OBJECTIVES: Takayasu arteritis (TAK) is a large-vessel vasculitis of unknown aetiology. The annual incidence in hospital-based studies is 1-2/million. The UK General Practice Research Database (UKGPRD) contains complete primary care records on 3.6 million people. There are no data on the incidence of TAK in the UK or from primary care anywhere in the world. The aim of this study was to determine the annual incidence of TAK in the UK using the UKGPRD and in a well defined hospital population [Norfolk Vasculitis Register (NORVASC)]. METHODS: We identified all patients in the UKGPRD with a first diagnosis of TAK during 2000 05, using the Read code (G757); and in the NORVASC population. The annual incidence was calculated as the number of incident cases divided by total person years. RESULTS: A total of 14 (13 females) subjects were identified with a first diagnosis of TAK during 2000-05 in the UKGPRD. The median age was 51.0 years (interquartile range 28-66). The overall annual incidence of TAK was 0.8/million (95% CI 0.4, 1.3). The incidence was stable throughout the study period. The mean prevalence of TAK was 4.7/million. There were six patients (five females) aged <40 years presenting in 2000-05 with TAK. The annual incidence in those aged <40 years was 0.3/million. In the NORVASC population, one case was identified (0.4/million/year) with three prevalent cases (7.1/million). CONCLUSION: This is the first population-based study of the epidemiology of TAK. The annual incidence and prevalence are consistent with previous studies. PMID- 19542213 TI - Aspects relevant for functioning in patients with ankylosing spondylitis according to the health professionals: a Delphi study with the ICF as reference. AB - OBJECTIVE: In AS there is no agreed definition of which aspects are important when describing functioning. This limits the possibility to classify, evaluate and investigate the consequences of the disease. This study aimed to achieve consensus among health professionals on which aspects of functioning are typical and relevant for AS patients using the International Classification of Functioning, Disability and Health (ICF) as reference. METHODS: An international Delphi study through e-mail was performed among different health professions. Answers to open questions on areas relevant for functioning in the first round were linked to ICF categories and analysed in the two following two rounds for the degree of consensus. RESULTS: Of the 267 experts invited, 126 agreed to participate and 74 participated in all rounds; 28 were rheumatologists, 6 rheumatology nurses, 24 physiotherapists, 2 occupational therapists, 4 psychologists, 8 rehabilitation physicians and 2 social workers. More than 80% agreement was reached on 141 ICF categories, of which 30 (21%) were part of Body functions; 27 (19%) of Body structures; 56 (40%) of Activities and Participation; and 28 (20%) of Environmental factors. In addition, two Personal factors-illness knowledge and coping-were agreed upon. CONCLUSION: 141 ICF categories and two personal factors represent the reference of functioning in AS from the perspective of health professional. The largest number of categories concerned restrictions in activities. Also, the impact of AS on participation in life situations and the role of environmental factors were underscored. This broadens the view on functioning in AS and has implications for future research into functioning. PMID- 19542214 TI - Immunohistochemical detection of intravascular platelet microthrombi in patients with lupus nephritis and anti-phospholipid antibodies. AB - OBJECTIVES: To evaluate whether the use of platelet immunohistochemistry (IHC) markers improves the sensitivity of histological methods to detect microthrombosis in SLE nephritis and aPLs and to analyse the clinicopathological correlations of microthrombosis in this setting. METHODS: Kidney biopsy specimens from 65 patients with SLE, including 36 with positive aPLs, were studied by IHC using antibodies against platelet glycoproteins CD41 and CD61. Clinical data at the time of kidney biopsy and during a mean follow-up of 7.5 years after biopsy were recorded and analysed with regard to histological or IHC data. RESULTS: Histological lesions previously defined as APS nephropathy were found in 33% of the SLE kidney biopsies and were not associated with positive aPLs. Microthrombi detected as intravascular CD61(+) platelet deposits were present in 43% of the tissues and were significantly associated with positive aPLs, but not with histological APS nephropathy, nephritis manifestations nor with renal outcome. Histological APS lesions but not CD61(+) microthrombi correlated with an older age at nephritis presentation, previous cardiovascular risk factors and worse renal outcome. CONCLUSIONS: Immunodetection of intravascular CD61(+) platelet aggregates is more sensitive than histological evaluation to detect acute microthrombosis and provides a better correlation with aPLs in SLE patients. In contrast, histological lesions consistent with APS nephropathy were not associated with aPLs but with cardiovascular risk factors and worse renal outcome. PMID- 19542215 TI - Expression of advanced glycation end products and their receptor in skin from patients with systemic sclerosis with and without calcinosis. AB - OBJECTIVES: Our aim was to establish which tissue components express advanced glycation/lipoperoxidation end products (AGEs) and their receptor (RAGE) in skin from patients with SSc, and how their expression relates to the disease subtypes and various clinical parameters. METHODS: Skin punch biopsies were taken from the forearms of 61 SSc patients with lcSSc; 32 with calcinosis (lcSScCal) and 29 without lcSSc, 36 with the dcSSc subtype and 22 healthy control subjects. Immunohistochemical localization of AGE-CML [N(epsilon)-(carboxymethyl) lysine] and RAGE was assessed semi-quantitatively on the microvascular endothelium, dermal fibroblasts and the cutaneous extracellular matrix (ECM). The Kruskal Wallis one-way ANOVA was used to compare data between groups. RESULTS: AGE-CML expression on the papillary dermis ECM of lcSScCal was greater than in the control group (P = 0.016). The reticular dermis of lcSScCal showed increased AGE N(epsilon)-(carboxymethyl) lysine (CML) expression compared with controls (P = 0.002), dcSSc (P = 0.024) and lcSSc (P = 0.025). Increased immunostaining for RAGE was seen on the reticular dermis ECM of the lcSScCal group compared with controls (P = 0.007). The lcSScCal subgroup showed statistically significant correlations for AGE-CML, and to a lesser extent for RAGE, with increased RP duration. There was no consistent evidence that the expression of AGE-CML or RAGE related to autoantibody status, clinical or histological skin score or patient age. CONCLUSIONS: Our results indicate the possible contribution of AGE-CML deposition on the ECM in the dermis of the lcSScCal subgroup to the pathogenesis of formation of calcinotic deposits. PMID- 19542216 TI - PGC-1{alpha} and PGC-1{beta} regulate mitochondrial density in neurons. AB - Recent studies indicate that regulation of cellular oxidative capacity through enhancing mitochondrial biogenesis may be beneficial for neuronal recovery and survival in human neurodegenerative disorders. The peroxisome proliferator activated receptor gamma coactivator-1alpha (PGC-1alpha) has been shown to be a master regulator of mitochondrial biogenesis and cellular energy metabolism in muscle and liver. The aim of our study was to establish whether PGC-1alpha and PGC-1beta control mitochondrial density also in neurons and if these coactivators could be up-regulated by deacetylation. The results demonstrate that PGC-1alpha and PGC-1beta control mitochondrial capacity in an additive and independent manner. This effect was observed in all studied subtypes of neurons, in cortical, midbrain, and cerebellar granule neurons. We also observed that endogenous neuronal PGC-1alpha but not PGC-1beta could be activated through its repressor domain by suppressing it. Results demonstrate also that overexpression of SIRT1 deacetylase or suppression of GCN5 acetyltransferase activates transcriptional activity of PGC-1alpha in neurons and increases mitochondrial density. These effects were mediated exclusively via PGC-1alpha, since overexpression of SIRT1 or suppression of GCN5 was ineffective where PGC-1alpha was suppressed by short hairpin RNA. Moreover, the results demonstrate that overexpression of PGC-1beta or PGC-1alpha or activation of the latter by SIRT1 protected neurons from mutant alpha-synuclein- or mutant huntingtin-induced mitochondrial loss. These evidences demonstrate that activation or overexpression of the PGC-1 family of coactivators could be used to compensate for neuronal mitochondrial loss and suggest that therapeutic agents activating PGC-1 would be valuable for treating neurodegenerative diseases in which mitochondrial dysfunction and oxidative damage play an important pathogenic role. PMID- 19542218 TI - Aqueous accessibility to the transmembrane regions of subunit c of the Escherichia coli F1F0 ATP synthase. AB - Rotary catalysis in F(1)F(0) ATP synthase is powered by proton translocation through the membrane-embedded F(0) sector. Proton binding and release occur in the middle of the membrane at Asp-61 on transmembrane helix (TMH) 2 of subunit c. Previously the reactivity of Cys substituted into TMH2 revealed extensive aqueous access at the cytoplasmic side as probed with Ag(+) and other thiolate-directed reagents. The analysis of aqueous accessibility of membrane-embedded regions in subunit c was extended here to TMH1 and the periplasmic side of TMH2. The Ag(+) sensitivity of Cys substitutions was more limited on the periplasmic versus cytoplasmic side of TMH2. In TMH1, Ag(+) sensitivity was restricted to a pocket of four residues lying directly behind Asp-61. Aqueous accessibility was also probed using Cd(2+), a membrane-impermeant soft metal ion with properties similar to Ag(+). Cd(2+) inhibition was restricted to the I28C substitution in TMH1 and residues surrounding Asp-61 in TMH2. The overall pattern of inhibition, by all of the reagents tested, indicates highest accessibility on the cytoplasmic side of TMH2 and in a pocket of residues around Asp-61, including proximal residues in TMH1. Additionally subunit a was shown to mediate access to this region by the membrane-impermeant probe 2-(trimethylammonium)ethyl methanethiosulfonate. Based upon these results and other information, a pocket of aqueous accessible residues, bordered by the peripheral surface of TMH4 of subunit a, is proposed to extend from the cytoplasmic side of cTMH2 to Asp-61 in the center of the membrane. PMID- 19542217 TI - Antigen-specific proteolysis by hybrid antibodies containing promiscuous proteolytic light chains paired with an antigen-binding heavy chain. AB - The antigen recognition site of antibodies consists of the heavy and light chain variable domains (V(L) and V(H) domains). V(L) domains catalyze peptide bond hydrolysis independent of V(H) domains (Mei, S., Mody, B., Eklund, S. H., and Paul, S. (1991) J. Biol. Chem. 266, 15571-15574). V(H) domains bind antigens noncovalently independent of V(L) domains (Ward, E. S., Gussow, D., Griffiths, A. D., Jones, P. T., and Winter, G. (1989) Nature 341, 544-546). We describe specific hydrolysis of fusion proteins of the hepatitis C virus E2 protein with glutathione S-transferase (GST-E2) or FLAG peptide (FLAG-E2) by antibodies containing the V(H) domain of an anti-E2 IgG paired with promiscuously catalytic V(L) domains. The hybrid IgG hydrolyzed the E2 fusion proteins more rapidly than the unpaired light chain. An active site-directed inhibitor of serine proteases inhibited the proteolytic activity of the hybrid IgG, indicating a serine protease mechanism. The hybrid IgG displayed noncovalent E2 binding in enzyme linked immunosorbent assay tests. Immunoblotting studies suggested hydrolysis of FLAG-E2 at a bond within E2 located approximately 11 kDa from the N terminus. GST E2 was hydrolyzed by the hybrid IgG at bonds in the GST tag. The differing cleavage pattern of FLAG-E2 and GST-E2 can be explained by the split-site model of catalysis, in which conformational differences in the E2 fusion protein substrates position alternate peptide bonds in register with the antibody catalytic subsite despite a common noncovalent binding mechanism. These studies provide proof-of-principle that the catalytic activity of a light chain can be rendered antigen-specific by pairing with a noncovalently binding heavy chain subunit. PMID- 19542219 TI - Structure of insoluble rat sperm glyceraldehyde-3-phosphate dehydrogenase (GAPDH) via heterotetramer formation with Escherichia coli GAPDH reveals target for contraceptive design. AB - Sperm glyceraldehyde-3-phosphate dehydrogenase has been shown to be a successful target for a non-hormonal contraceptive approach, but the agents tested to date have had unacceptable side effects. Obtaining the structure of the sperm-specific isoform to allow rational inhibitor design has therefore been a goal for a number of years but has proved intractable because of the insoluble nature of both native and recombinant protein. We have obtained soluble recombinant sperm glyceraldehyde-3-phosphate dehydrogenase as a heterotetramer with the Escherichia coli glyceraldehyde-3-phosphate dehydrogenase in a ratio of 1:3 and have solved the structure of the heterotetramer which we believe represents a novel strategy for structure determination of an insoluble protein. A structure was also obtained where glyceraldehyde 3-phosphate binds in the P(s) pocket in the active site of the sperm enzyme subunit in the presence of NAD. Modeling and comparison of the structures of human somatic and sperm-specific glyceraldehyde-3-phosphate dehydrogenase revealed few differences at the active site and hence rebut the long presumed structural specificity of 3-chlorolactaldehyde for the sperm isoform. The contraceptive activity of alpha-chlorohydrin and its apparent specificity for the sperm isoform in vivo are likely to be due to differences in metabolism to 3-chlorolactaldehyde in spermatozoa and somatic cells. However, further detailed analysis of the sperm glyceraldehyde-3-phosphate dehydrogenase structure revealed sites in the enzyme that do show significant difference compared with published somatic glyceraldehyde-3-phosphate dehydrogenase structures that could be exploited by structure-based drug design to identify leads for novel male contraceptives. PMID- 19542220 TI - Mirk regulates the exit of colon cancer cells from quiescence. AB - Mirk/Dyrk1B is a serine/threonine kinase widely expressed in colon cancers. Serum starvation induced HD6 colon carcinoma cells to enter a quiescent G0 state, characterized by a 2N DNA content and a lower RNA content than G1 cells. Compared with cycling cells, quiescent cells exhibited 16-fold higher levels of the retinoblastoma protein p130/Rb2, which sequesters E2F4 to block entry into G1, 10 fold elevated levels of the CDK inhibitor p27kip1, and 10-fold higher levels of Mirk. However, depletion of Mirk did not prevent entry into G0, but enabled quiescent HD6, SW480, and colo320 colon carcinoma cells to acquire some biochemical characteristics of G1 cells, including increased levels of cyclin D1 and cyclin D3 because of slower turnover, increased activity of their CDK4/cyclin D complexes, and increased phosphorylation and decreased E2F4 sequestering ability of the CDK4 target, p130/Rb2. As a result, depletion of Mirk allowed some cells to escape quiescence and enabled cells released from quiescence to traverse G1 more quickly. The kinase activity of Mirk was increased by the chemotherapeutic drug 5-fluorouracil (5-FU). Treatment of p53 mutant colon cancer cells with 5-FU led to an elongated G1 in a Mirk-dependent manner, as G1 was shortened by ectopic overexpression of cyclin D1 mutated at the Mirk phosphorylation site (T288A), but not by wild-type cyclin D1. Mirk, through regulating cyclin D turnover, and the CDK inhibitor p27, as shown by depletion studies, functioned independently and additively to regulate the exit of tumor cells from quiescence. PMID- 19542221 TI - Differential processing of amyloid-beta precursor protein directs human embryonic stem cell proliferation and differentiation into neuronal precursor cells. AB - The amyloid-beta precursor protein (AbetaPP) is a ubiquitously expressed transmembrane protein whose cleavage product, the amyloid-beta (Abeta) protein, is deposited in amyloid plaques in neurodegenerative conditions such as Alzheimer disease, Down syndrome, and head injury. We recently reported that this protein, normally associated with neurodegenerative conditions, is expressed by human embryonic stem cells (hESCs). We now report that the differential processing of AbetaPP via secretase enzymes regulates the proliferation and differentiation of hESCs. hESCs endogenously produce amyloid-beta, which when added exogenously in soluble and fibrillar forms but not oligomeric forms markedly increased hESC proliferation. The inhibition of AbetaPP cleavage by beta-secretase inhibitors significantly suppressed hESC proliferation and promoted nestin expression, an early marker of neural precursor cell (NPC) formation. The induction of NPC differentiation via the non-amyloidogenic pathway was confirmed by the addition of secreted AbetaPPalpha, which suppressed hESC proliferation and promoted the formation of NPCs. Together these data suggest that differential processing of AbetaPP is normally required for embryonic neurogenesis. PMID- 19542222 TI - Paradoxical condensation of copper with elevated beta-amyloid in lipid rafts under cellular copper deficiency conditions: implications for Alzheimer disease. AB - Redox-active copper is implicated in the pathogenesis of Alzheimer disease (AD), beta-amyloid peptide (Abeta) aggregation, and amyloid formation. Abeta.copper complexes have been identified in AD and catalytically oxidize cholesterol and lipid to generate H2O2 and lipid peroxides. The site and mechanism of this abnormality is not known. Growing evidence suggests that amyloidogenic processing of the beta-amyloid precursor protein (APP) occurs in lipid rafts, membrane microdomains enriched in cholesterol. beta- and gamma-secretases, and Abeta have been identified in lipid rafts in cultured cells, human and rodent brains, but the role of copper in lipid raft amyloidogenic processing is presently unknown. In this study, we found that copper modulates flotillin-2 association with cholesterol-rich lipid raft domains, and consequently Abeta synthesis is attenuated via copper-mediated inhibition of APP endocytosis. We also found that total cellular copper is associated inversely with lipid raft copper levels, so that under intracellular copper deficiency conditions, Abeta.copper complexes are more likely to form. This explains the paradoxical hypermetallation of Abeta with copper under tissue copper deficiency conditions in AD. PMID- 19542223 TI - Lactogenic hormonal induction of long distance interactions between beta-casein gene regulatory elements. AB - Lactogenic hormone regulation of beta-casein gene expression in mammary epithelial cells provides an excellent model in which to study the mechanisms by which steroid and peptide hormone signaling control gene expression. Prolactin- and glucocorticoid-mediated induction of beta-casein gene expression involves two principal regulatory regions, a proximal promoter and a distal enhancer located in the mouse approximately -6 kb upstream of the transcription start site. Using a chromosome conformation capture assay and quantitative real time PCR, we demonstrate that a chromatin loop is created in conjunction with the recruitment of specific transcription factors and p300 in HC11 mammary epithelial cells. Stimulation with both prolactin and hydrocortisone is required for the induction of these long range interactions between the promoter and enhancer, and no DNA looping was observed in nontreated cells or cells treated with each of the hormones separately. The lactogenic hormone-induced interaction between the proximal promoter and distal enhancer was confirmed in hormone-treated primary three-dimensional mammary acini cultures. In addition, the developmental regulation of DNA looping between the beta-casein regulatory regions was observed in lactating but not in virgin mouse mammary glands. Furthermore, beta-casein mRNA induction and long range interactions between these regulatory regions were inhibited in a progestin-dependent manner following stimulation with prolactin and hydrocortisone in HC11 cells expressing human PR-B. Collectively, these data suggest that the communication between these regulatory regions with intervening DNA looping is a crucial step required to both create and maintain active chromatin domains and regulate transcription. PMID- 19542224 TI - The first draft of the endostatin interaction network. AB - Endostatin is a C-terminal proteolytic fragment of collagen XVIII that is localized in vascular basement membrane zones in various organs. It binds to heparin/heparan sulfate and to a number of proteins, but its molecular mechanisms of action are not fully elucidated. We have used surface plasmon resonance (SPR) arrays to identify new partners of endostatin, and to give further insights on its molecular mechanism of action. New partners of endostatin include glycosaminoglycans (chondroitin and dermatan sulfate), matricellular proteins (thrombospondin-1 and SPARC), collagens (I, IV, and VI), the amyloid peptide Abeta-(1-42), and transglutaminase-2. The biological functions of the endostatin network involve a number of extracellular proteins containing epidermal growth factor and epidermal growth factor-like domains, and able to bind calcium. Depending on the trigger event, and on the availability of its members in a given tissue at a given time, the endostatin network might be involved either in the control of angiogenesis, and tumor growth, or in neurogenesis and neurodegenerative diseases. PMID- 19542225 TI - Modulation of carcinogen metabolism by nitric oxide-aspirin 2 is associated with suppression of DNA damage and DNA adduct formation. AB - Nitric oxide (NO)-donating non-steroidal anti-inflammatory drugs (NSAIDs) represent a promising new class of drugs developed to provide a safer alternative than their conventional NSAID counterparts in chemoprevention. We tested the effects of NO-aspirin 2 on Phase I and Phase II carcinogen-metabolizing enzymes. In HepG2 human hepatoma cells and in LS180 colonic adenocarcinoma cells, NO aspirin 2 inhibited 2,3,7,8-tetrachlordibenzo-p-dioxin (TCDD)-induced cytochrome P450 (CYP) enzyme activity and CYP1A1 and CYP1A2 mRNA expression. These effects were further characterized as being mediated through transcriptional regulation: NO-aspirin 2 inhibited binding of ligand (TCDD)-activated aryl hydrocarbon receptor to the CYP1A1 enhancer sequence; additionally, NO-aspirin 2 suppressed carcinogen-induced expression of CYP1A heterogeneous nuclear RNA. The fate of carcinogen metabolites depends not only on activation by CYP enzymes but also detoxification by Phase II enzymes. Both HepG2 and LS180 cells treated with NO aspirin 2 showed an increase in glutathione S-transferase-P1 (GST-P1), glutamate cysteine ligase (GCL), and NAD(P)H:quinone oxidoreductase-1 (NQO1) expression. Compared with two other NO-releasing compounds, diethylenetriamine-NO and the organic nitrate, isosorbide dinitrate, the inhibitory effects of NO-aspirin 2 on TCDD-induced CYP activity and mRNA expression were considerably more potent. Furthermore, aspirin alone had no inhibitory effect on TCDD-induced CYP activity, nor did aspirin up-regulate GCL, GST-P1, or NQO1 expression. Consequent to the effects on carcinogen-metabolizing enzymes, NO-aspirin 2 inhibited [3H]benzo[a]pyrene-DNA adduct formation and DNA damage elicited by TCDD or benzo[a]pyrene. Our results demonstrate that NO-aspirin 2 may be an effective chemopreventive agent by favorably affecting the inhibitory and enhancing effects of Phase I and Phase II carcinogen metabolism, thereby protecting DNA from carcinogenic insult. PMID- 19542226 TI - Conserved active site sequences in Arabidopsis plastid terminal oxidase (PTOX): in vitro and in planta mutagenesis studies. AB - The plastid terminal oxidase (PTOX) is distantly related to the mitochondrial alternative oxidase (AOX). Both are members of the diiron carboxylate quinol oxidase (DOX) class of proteins. PTOX and AOX contain 20 highly conserved amino acids, six of which are Fe-binding ligands. We have previously used in vitro and in planta activity assays to examine the functional importance of the Fe-binding sites. In this report, we conduct alanine-scanning mutagenesis on the 14 other conserved sites using our in vitro and in planta assay procedures. We found that the 14 sites fall into three classes: (i) Ala-139, Pro-142, Glu-171, Asn-174, Leu 179, Pro-216, Ala-230, Asp-287, and Arg-293 are dispensable for activity; (ii) Tyr-234 and Asp-295 are essential for activity; and (iii) Leu-135, His-151, and Tyr-212 are important but not essential for activity. Our data are consistent with the proposed role of some of these residues in active site conformation, substrate binding, and/or catalysis. Titration experiments showed that down regulation of PTOX to approximately 3% of wild-type levels did not compromise plant growth, at least under ambient growth conditions. This suggests that PTOX is normally in excess, especially early in thylakoid membrane biogenesis. PMID- 19542227 TI - LFA-1-dependent Ca2+ entry following suboptimal T cell receptor triggering proceeds without mobilization of intracellular Ca2+. AB - A surge in cytosolic calcium ion concentration by entry of extracellular Ca2+ is a hallmark of T cell activation. According to store-operated Ca2+ entry mechanism, the Ca2+ entry is preceded by activation of phospholipase C-gamma1 (PLC-gamma1) and the consequent mobilization of intracellular Ca2+. Using membrane vesicles expressing the mouse class I major histocompatibility complex, i.e. Ld plus costimulatory ligands, i.e. B7-1 and intercellular adhesion molecule 1 along with 2C T cell receptor transgenic T cells, we investigated the roles of CD28 and LFA-1 (lymphocyte function-associated antigen-1) in the activation of PLC-gamma1 and Ca2+ signaling. Both CD28 and LFA-1 made significant and comparable contributions to the activation of PLC-gamma1 as gauged by the level of its phosphorylation at tyrosine 783. In contrast, their roles in Ca2+ signaling were quite distinct so that LFA-1/intercellular adhesion molecule-1 interaction exerted a determining role, whereas CD28/B7-1 interaction played only a minimal role. In particular, when the T cells were activated by suboptimal T cell receptor stimulation, LFA-1 played an indispensable role in the Ca2+ signaling. Further experiments using Ca2+-free medium demonstrated that the entry of extracellular Ca2+ was not always accompanied by mobilization of intracellular Ca2+. Thus, intracellular Ca2+ mobilization was hardly detected under the condition that LFA-1 played the indispensable role in the entry of extracellular Ca2+, while a distinct level of intracellular Ca2+ mobilization was readily detected under the condition that LFA-1 played only the supporting role. These results ensure the unique role of LFA-1 in T cell Ca2+ signaling and reveal that LFA-1-dependent Ca2+ entry proceeds via a mechanism separate from store-operated Ca2+ entry. PMID- 19542228 TI - Structural and functional elucidation of the mechanism promoting error-prone synthesis by human DNA polymerase kappa opposite the 7,8-dihydro-8-oxo-2' deoxyguanosine adduct. AB - Human polymerase kappa (hPol kappa) is one of four eukaryotic Y-class DNA polymerases and may be an important element in the cellular response to polycyclic aromatic hydrocarbons such as benzo[a]pyrene, which can lead to reactive oxygenated metabolite-mediated oxidative stress. Here, we present a detailed analysis of the activity and specificity of hPol kappa bypass opposite the major oxidative adduct 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxoG). Unlike its archaeal homolog Dpo4, hPol kappa bypasses this lesion in an error-prone fashion by inserting mainly dATP. Analysis of transient-state kinetics shows diminished "bursts" for dATP:8-oxoG and dCTP:8-oxoG incorporation, indicative of non-productive complex formation, but dATP:8-oxoG insertion events that do occur are 2-fold more efficient than dCTP:G insertion events. Crystal structures of ternary hPol kappa complexes with adducted template-primer DNA reveal non productive (dGTP and dATP) alignments of incoming nucleotide and 8-oxoG. Structural limitations placed upon the hPol kappa by interactions between the N clasp and finger domains combined with stabilization of the syn-oriented template 8-oxoG through the side chain of Met-135 both appear to contribute to error-prone bypass. Mutating Leu-508 in the little finger domain of hPol kappa to lysine modulates the insertion opposite 8-oxoG toward more accurate bypass, similar to previous findings with Dpo4. Our structural and activity data provide insight into important mechanistic aspects of error-prone bypass of 8-oxoG by hPol kappa compared with accurate and efficient bypass of the lesion by Dpo4 and polymerase eta. PMID- 19542229 TI - The elucidation of the structure of Thermotoga maritima peptidoglycan reveals two novel types of cross-link. AB - Thermotoga maritima is a Gram-negative, hyperthermophilic bacterium whose peptidoglycan contains comparable amounts of L- and D-lysine. We have determined the fine structure of this cell-wall polymer. The muropeptides resulting from the digestion of peptidoglycan by mutanolysin were separated by high-performance liquid chromatography and identified by amino acid analysis after acid hydrolysis, dinitrophenylation, enzymatic determination of the configuration of the chiral amino acids, and mass spectrometry. The high-performance liquid chromatography profile contained four main peaks, two monomers, and two dimers, plus a few minor peaks corresponding to anhydro forms. The first monomer was the d-lysine-containing disaccharide-tripeptide in which the D-Glu-D-Lys bond had the unusual gamma-->epsilon arrangement (GlcNAc-MurNAc-L-Ala-gamma-D-Glu-epsilon-D Lys). The second monomer was the conventional disaccharide-tetrapeptide (GlcNAc MurNAc-L-Ala-gamma-D-Glu-L-Lys-D-Ala). The first dimer contained a disaccharide-L Ala as the acyl donor cross-linked to the alpha-amine of D-Lys in a tripeptide acceptor stem with the sequence of the first monomer. In the second dimer, donor and acceptor stems with the sequences of the second and first monomers, respectively, were connected by a D-Ala4-alpha-D-Lys3 cross-link. The cross linking index was 10 with an average chain length of 30 disaccharide units. The structure of the peptidoglycan of T. maritima revealed for the first time the key role of D-Lys in peptidoglycan synthesis, both as a surrogate of L-Lys or meso diaminopimelic acid at the third position of peptide stems and in the formation of novel cross-links of the L-Ala1(alpha-->alpha)D-Lys3 and D-Ala4(alpha- >alpha)D-Lys3 types. PMID- 19542230 TI - Mechanism and regulatory function of CpG signaling via scavenger receptor B1 in primary B cells. AB - It is well established that CpG promotes pro-inflammatory cytokine and antibody production by B cells via the Toll-like receptor 9 (TLR9)-dependent pathway. However, scavenger receptors (SRs) are also capable of binding such pathogen derived molecules, yet their contribution to CpG-induced signaling events has not yet been evaluated. Here we identified a novel TLR9-independent mechanism of CpG induced signaling and immune function that is mediated by the scavenger B1 receptor (SR-B1). Specifically, we show that CpG/SR-B1 triggers calcium entry into primary B lymphocytes via phospholipase C gamma-1-mediated activation of TRPC3 channels and also B cell adhesion to vascular cell adhesion molecule-1. CpG induced calcium signals and vascular cell adhesion molecule-1 adhesion are TLR9 independent and are mediated exclusively by SR-B1. Although pro-inflammatory cytokine and Ig production induced by CpG require TLR9 expression, we also found that SR-B1 negatively regulates TLR9-dependent production of interleukin-6, interleukin-10, and IgM. Thus, our results provide a novel perspective on the complexity of CpG signaling within B cells by demonstrating that SR-B1 is an alternative pathway for nucleic acid-induced signaling that provides feedback inhibition on specific TLR9-dependent responses of B cells. Consequently, these results have wide implications for understanding the mechanisms regulating immune tolerance to nucleic acids and pathogen-associated molecules. PMID- 19542231 TI - Requirement of myosin Vb.Rab11a.Rab11-FIP2 complex in cholesterol-regulated translocation of NPC1L1 to the cell surface. AB - Niemann-Pick C1-like 1 (NPC1L1) plays a critical role in the enterohepatic absorption of free cholesterol. Cellular cholesterol depletion induces the transport of NPC1L1 from the endocytic recycling compartment to the plasma membrane (PM), and cholesterol replenishment causes the internalization of NPC1L1 together with cholesterol via clathrin-mediated endocytosis. Although NPC1L1 has been characterized, the other proteins involved in cholesterol absorption and the endocytic recycling of NPC1L1 are largely unknown. Most of the vesicular trafficking events are dependent on the cytoskeleton and motor proteins. Here, we investigated the roles of the microfilament and microfilament-associated triple complex composed of myosin Vb, Rab11a, and Rab11-FIP2 in the transport of NPC1L1 from the endocytic recycling compartment to the PM. Interfering with the dynamics of the microfilament by pharmacological treatment delayed the transport of NPC1L1 to the cell surface. Meanwhile, inactivation of any component of the myosin Vb.Rab11a.Rab11-FIP2 triple complex inhibited the export of NPC1L1. Expression of the dominant-negative mutants of myosin Vb, Rab11a, or Rab11-FIP2 decreased the cellular cholesterol uptake by blocking the transport of NPC1L1 to the PM. These results suggest that the efficient transport of NPC1L1 to the PM is dependent on the microfilament-associated myosin Vb.Rab11a.Rab11-FIP2 triple complex. PMID- 19542232 TI - Activation of Cu,Zn-superoxide dismutase in the absence of oxygen and the copper chaperone CCS. AB - Eukaryotic Cu,Zn-superoxide dismutases (SOD1s) are generally thought to acquire the essential copper cofactor and intramolecular disulfide bond through the action of the CCS copper chaperone. However, several metazoan SOD1s have been shown to acquire activity in vivo in the absence of CCS, and the Cu,Zn-SOD from Caenorhabditis elegans has evolved complete independence from CCS. To investigate SOD1 activation in the absence of CCS, we compared and contrasted the CCS independent activation of C. elegans and human SOD1 to the strict CCS-dependent activation of Saccharomyces cerevisiae SOD1. Using a yeast expression system, both pathways were seen to acquire copper derived from cell surface transporters and compete for the same intracellular pool of copper. Like CCS, CCS-independent activation occurs rapidly with a preexisting pool of apo-SOD1 without the need for new protein synthesis. The two pathways, however, strongly diverge when assayed for the SOD1 disulfide. SOD1 molecules that are activated without CCS exhibit disulfide oxidation in vivo without oxygen and under copper-depleted conditions. The strict requirement for copper, oxygen, and CCS in disulfide bond oxidation appears exclusive to yeast SOD1, and we find that a unique proline at position 144 in yeast SOD1 is responsible for this disulfide effect. CCS dependent and -independent pathways also exhibit differential requirements for molecular oxygen. CCS activation of SOD1 requires oxygen, whereas the CCS independent pathway is able to activate SOD1s even under anaerobic conditions. In this manner, Cu,Zn-SOD from metazoans may retain activity over a wide range of physiological oxygen tensions. PMID- 19542233 TI - Hyperphosphorylation and aggregation of Tau in laforin-deficient mice, an animal model for Lafora disease. AB - Lafora progressive myoclonous epilepsy (Lafora disease; LD) is caused by mutations in the EPM2A gene encoding a dual specificity protein phosphatase named laforin. Our analyses on the Epm2a gene knock-out mice, which developed most of the symptoms of LD, reveal the presence of hyperphosphorylated Tau protein (Ser(396) and Ser(202)) as neurofibrillary tangles (NFTs) in the brain. Intriguingly, NFTs were also observed in the skeletal muscle tissues of the knock out mice. The hyperphosphorylation of Tau was associated with increased levels of the active form of GSK3 beta. The observations on Tau protein were replicated in cell lines using laforin overexpression and knockdown approaches. We also show here that laforin and Tau proteins physically interact and that the interaction was limited to the phosphatase domain of laforin. Finally, our in vitro and in vivo assays demonstrate that laforin dephosphorylates Tau, and therefore laforin is a novel Tau phosphatase. Taken together, our study suggests that laforin is one of the critical regulators of Tau protein, that the NFTs could underlie some of the symptoms seen in LD, and that laforin can contribute to the NFT formation in Alzheimer disease and other tauopathies. PMID- 19542234 TI - Purification and functional reconstitution of monomeric mu-opioid receptors: allosteric modulation of agonist binding by Gi2. AB - Despite extensive characterization of the mu-opioid receptor (MOR), the biochemical properties of the isolated receptor remain unclear. In light of recent reports, we proposed that the monomeric form of MOR can activate G proteins and be subject to allosteric regulation. A mu-opioid receptor fused to yellow fluorescent protein (YMOR) was constructed and expressed in insect cells. YMOR binds ligands with high affinity, displays agonist-stimulated [(35)S]guanosine 5'-(gamma-thio)triphosphate binding to Galpha(i), and is allosterically regulated by coupled G(i) protein heterotrimer both in insect cell membranes and as purified protein reconstituted into a phospholipid bilayer in the form of high density lipoprotein particles. Single-particle imaging of fluorescently labeled receptor indicates that the reconstituted YMOR is monomeric. Moreover, single-molecule imaging of a Cy3-labeled agonist, [Lys(7), Cys(8)]dermorphin, illustrates a novel method for studying G protein-coupled receptor-ligand binding and suggests that one molecule of agonist binds per monomeric YMOR. Together these data support the notion that oligomerization of the mu-opioid receptor is not required for agonist and antagonist binding and that the monomeric receptor is the minimal functional unit in regard to G protein activation and strong allosteric regulation of agonist binding by G proteins. PMID- 19542235 TI - MocA is a specific cytidylyltransferase involved in molybdopterin cytosine dinucleotide biosynthesis in Escherichia coli. AB - We have purified and characterized a specific CTP:molybdopterin cytidylyltransferase for the biosynthesis of the molybdopterin (MPT) cytosine dinucleotide (MCD) cofactor in Escherichia coli. The protein, named MocA, shows 22% amino acid sequence identity to E. coli MobA, the specific GTP:molybdopterin guanylyltransferase for molybdopterin guanine dinucleotide biosynthesis. MocA is essential for the activity of the MCD-containing enzymes aldehyde oxidoreductase YagTSR and the xanthine dehydrogenases XdhABC and XdhD. Using a fully defined in vitro assay, we showed that MocA, Mo-MPT, CTP, and MgCl2 are required and sufficient for MCD biosynthesis in vitro. The activity of MocA is specific for CTP; other nucleotides such as ATP and GTP were not utilized. In the defined in vitro system a turnover number of 0.37+/-0.01 min(-1) was obtained. A 1:1 binding ratio of MocA to Mo-MPT and CTP was determined to monomeric MocA with dissociation constants of 0.23+/-0.02 microm for CTP and 1.17+/-0.18 microm for Mo-MPT. We showed that MocA was also able to convert MPT to MCD in the absence of molybdate, however, with only one catalytic turnover. The addition of molybdate after one turnover gave rise to a higher MCD production, revealing that MCD remains bound to MocA in the absence of molybdate. This work presents the first characterization of a specific enzyme involved in MCD biosynthesis in bacteria. PMID- 19542236 TI - Zebrafish and mouse alpha2,3-sialyltransferases responsible for synthesizing GM4 ganglioside. AB - We have previously reported that fish pathogens causing vibriosis specifically adhere to GM4 on the epithelial cells of fish intestinal tracts (Chisada, S., Horibata, Y., Hama, Y., Inagaki, M., Furuya, N., Okino, N., and Ito, M. (2005) Biochem. Biophys. Res. Commun. 333, 367-373). To identify the gene encoding the enzyme for GM4 synthesis in the fish intestinal tract, a phylogenetic tree of vertebrate ST3GalVs, including Danio rerio and Oryzias latipes, was generated in which two putative subfamilies of fish ST3GalVs were found. Two putative ST3GalVs of zebrafish (zST3GalV-1 and -2), each belonging to different subfamilies, were cloned from the zebrafish cDNA library. Interestingly, zST3GalV-1 synthesized GM3 (NeuAcalpha2-3Galbeta1-4Glcbeta1-1'Cer) but not GM4, whereas zSTGalV-2 synthesized both gangliosides in vitro when expressed in CHO-K1 and RPMI1846 cells. Flow cytometric analysis using anti-GM4 antibody revealed that the transformation of RPMI1846 cells with zST3GalV-2 but not zST3GalV-1 cDNA increased the cell-surface expression of GM4. Whole mount in situ hybridization showed that the zST3GalV-2 transcript was strongly expressed in the gastrointestinal tract, whereas zST3GalV-1 was expressed in the brain and esophagus but not gastrointestinal tract in 3-day post-fertilization embryos. It has long been a matter of controversy which enzyme is responsible for the synthesis of GM4 in mammals. We found that three isoforms of mouse ST3GalV (mST3GalV) having different N-terminal sequences can synthesize GM4 as well as GM3 when expressed in RPMI1846 and CHO-K1 cells. Furthermore, mST3GalV knock-out mice were found to lack GM4 synthase activity and GM4 in contrast to wild-type mice. These results clearly indicate that zST3GalV-2 and mST3GalV are the enzymes responsible for the synthesis of GM4 in zebrafish and mice, respectively. PMID- 19542238 TI - Long-term benefits of testosterone replacement therapy on angina threshold and atheroma in men. AB - INTRODUCTION: In short-term studies, testosterone replacement therapy has been shown to protect male subjects from exercise-induced ischaemia and modify cardiovascular risk factors such as insulin resistance, fat mass and lipid profiles. METHODS: This randomised parallel group controlled trial was designed to assess the treatment effect of testosterone therapy (Nebido) compared with placebo in terms of exercise-induced ischaemia, lipid profiles, carotid intima media thickness (CIMT) and body composition during 12 months treatment in men with low testosterone levels and angina. RESULTS: A total of 15 men were recruited but 13 (n=13) reached adequate duration of follow-up; seven were treated with testosterone and six with placebo. Testosterone increased time to ischaemia (129+/-48 s versus 12+/-18, P=0.02) and haemoglobin (0.4+/-0.6 g/dl versus -0.03+/-0.5, P=0.04), and reduced body mass index (-0.3 kg/m(2) versus 1.3+/-1, P=0.04) and triglycerides (-0.36+/-0.4 mmol/l versus 0.3+/-1.2, P=0.05). The CIMT decreased in the testosterone group more than placebo, but full between group analyses suggested this was only a statistical trend (-0.5+/-0.1 vs -0.09+/ 0.06, P=0.16). There were no significant effects on serum prostate specific antigen, total or high-density lipoprotein cholesterol; or on mood and symptom scores as assessed by Seattle Angina Score and EuroQol. CONCLUSION: The protective effect of testosterone on myocardial ischaemia is maintained throughout treatment without decrement. Previously noted potentially beneficial effects of testosterone on body composition were confirmed and there were no adverse effects. PMID- 19542239 TI - Low prevalence of the metabolic syndrome but high occurrence of various metabolic disorders in Chinese women with polycystic ovary syndrome. AB - OBJECTIVE: Variations in the prevalence of metabolic syndrome (MetS) among women with polycystic ovary syndrome (PCOS) in different races were reported. We sought to report this prevalence and its components in Chinese women with PCOS and compared these characteristics with healthy controls. DESIGN: Anthropometric measurements and biochemical parameters were evaluated in 578 PCOS patients diagnosed by the Rotterdam criteria and 281 age- and body mass index (BMI) matched controls. International Diabetes Federation criteria for MetS were used. RESULTS: The prevalence of MetS was 16.8% in this study, and 60.7% of patients displayed at least one component of MetS. Among the patients, the rates of dyslipidemia, impaired fasting glucose, and elevated blood pressure were 41.6, 19.8, and 16.1% respectively; the rates of these corresponding components in age- and BMI-matched controls were 14.6, 5.3, and 5.7% respectively. In PCOS patients, the prevalence of MetS was 0.0, 3.9, 20.2, and 51.1% for four different BMI groups respectively; the prevalence of MetS was 7.3, 14.9, 24.2, and 42.4% in the four age groups respectively. Nearly 90% of patients diagnosed with MetS belonged to overweight and obese groups. BMI and age rather than free testosterone, free androgen index, fasting insulin, or sex hormone-binding globulin were included in formulation for predicting MetS according to multivariable logistic regression. CONCLUSIONS: Low prevalence of MetS but high occurrence of various metabolic disorders was found in women with PCOS compared with age- and BMI-matched controls in this study. BMI and age appeared to contribute more to developing MetS than other parameters associated with insulin resistance or hyperandrogenism. PMID- 19542237 TI - Structure-function relationships in miscoding by Sulfolobus solfataricus DNA polymerase Dpo4: guanine N2,N2-dimethyl substitution produces inactive and miscoding polymerase complexes. AB - Previous work has shown that Y-family DNA polymerases tolerate large DNA adducts, but a substantial decrease in catalytic efficiency and fidelity occurs during bypass of N2,N2-dimethyl (Me2)-substituted guanine (N2,N2-Me2G), in contrast to a single methyl substitution. Therefore, it is unclear why the addition of two methyl groups is so disruptive. The presence of N2,N2-Me2G lowered the catalytic efficiency of the model enzyme Sulfolobus solfataricus Dpo4 16,000-fold. Dpo4 inserted dNTPs almost at random during bypass of N2,N2-Me2G, and much of the enzyme was kinetically trapped by an inactive ternary complex when N2,N2-Me2G was present, as judged by a reduced burst amplitude (5% of total enzyme) and kinetic modeling. One crystal structure of Dpo4 with a primer having a 3'-terminal dideoxycytosine (Cdd) opposite template N2,N2-Me2G in a post-insertion position showed Cdd folded back into the minor groove, as a catalytically incompetent complex. A second crystal had two unique orientations for the primer terminal Cdd as follows: (i) flipped into the minor groove and (ii) a long pairing with N2,N2 Me2G in which one hydrogen bond exists between the O-2 atom of Cdd and the N-1 atom of N2,N2-Me2G, with a second water-mediated hydrogen bond between the N-3 atom of Cdd and the O-6 atom of N2,N2-Me2G. A crystal structure of Dpo4 with dTTP opposite template N2,N2-Me2G revealed a wobble orientation. Collectively, these results explain, in a detailed manner, the basis for the reduced efficiency and fidelity of Dpo4-catalyzed bypass of N2,N2-Me2G compared with mono-substituted N2 alkyl G adducts. PMID- 19542240 TI - Thirst perception and arginine vasopressin production in a kindred with an activating mutation of the type 2 vasopressin receptor: the pathophysiology of nephrogenic syndrome of inappropriate antidiuresis. AB - BACKGROUND: Activating mutations of the vasopressin receptor gene on the X chromosome cause the nephrogenic syndrome of inappropriate antidiuresis (NSIAD). We describe a male child who presented with persistent hyponatraemia and whose mother was also found to be hyponatraemic. She had learnt to avoid excess fluid consumption because of associated malaise. Both individuals had a subnormal ability to excrete a water load with mother also demonstrating a heightened sense of thirst at low serum osmolalities. RESULTS: Mother and child were found to have the previously characterised activating mutation (p.Arg137Cys) of the arginine vasopressin receptor type 2 gene (AVPR2), but had measurable levels of AVP when hyponatraemic. CONCLUSIONS: We conclude that female carriers of activating mutations of the vasopressin receptor are susceptible to hyponatraemia and therefore need to be provided with advice regarding fluid intake. An altered thirst perception may increase susceptibility to hyponatraemia. We confirm that the presence of measurable amounts of AVP in patients with hyponatraemia does not exclude the diagnosis of NSIAD. PMID- 19542241 TI - Inflammatory mediators in morbidly obese subjects: associations with glucose abnormalities and changes after oral glucose. AB - OBJECTIVE: To explore inflammatory mediators in morbidly obese (MO) subjects with various categories of glucose tolerance and to study the changes in these mediators after an oral glucose load. DESIGN: Cross-sectional and experimental study. METHODS: A total of 144 MO subjects were classified into three categories: normal glucose tolerance (NGT); pre-diabetes; and new onset diabetes mellitus (NODM) were included, as were 27 normal weight normoglycemic controls. Serum osteoprotegerin (OPG), visfatin, leptin, adiponectin, interleukin-1 receptor antagonist (IL-1Ra), and C-reactive protein (CRP) were analyzed during an oral glucose tolerance test (OGTT). RESULTS: Fasting levels of leptin and IL-1Ra were consistently higher in obese persons (P<0.001 and P<0.05). MO subjects with NGT had higher CRP levels (P<0.001) and lower adiponectin levels (P<0.05) compared to controls. Yet when compared with MO subjects with NODM, those with NGT had lower CRP levels and higher adiponectin levels (both P<0.05). Baseline OPG and visfatin levels did not differ between the groups (P=0.326 and P=0.198). During OGTT, OPG levels decreased (P<0.001) and visfatin levels increased transiently (P=0.018). The response in OPG and visfatin did not differ between the groups (P=0.690 and P=0.170). There were minor changes in adiponectin and leptin levels. CONCLUSIONS: Morbid obesity and glucose intolerance were associated with lower adiponectin levels and higher CRP levels, thus supporting a relationship between obesity, glucose homeostasis, and inflammation. Oral glucose suppressed OPG levels and transiently enhanced visfatin levels independent of obesity and glucose tolerance status, indicating that glucose may be involved in the acute regulation of these proteins. PMID- 19542242 TI - Differences in expression and activity of 11beta-hydroxysteroid dehydrogenase type 1 and 2 in human placentas of term pregnancies according to birth weight and gender. AB - BACKGROUND: Fetal exposure to maternal glucocorticoids may determine fetal growth and the programing of later disorders. Availability of the glucocorticoids in the placenta is regulated by the 11beta-hydroxysteroid dehydrogenase (11beta-HSDs) enzymes. To date, there are discrepancies with regard to cortisol (F) cord blood levels in fetuses with intrauterine growth retardation in different species. Objective To study the expression and activity of 11beta-HSDs in placentas from full term small for gestational age (SGA), appropriate for gestational age (AGA) and large for gestational age (LGA) newborns, and cortisol cord blood concentration. METHODS: Twenty-five placentas from AGA, 24 SGA and 25 LGA were collected. RESULTS: SGA newborns had significantly lower and LGA newborns had significantly higher birth weight, birth length, head circumference, and placental weight than AGA counterparts. We observed a direct correlation between placental weight and birth weight, birth length and head circumference, and higher cord F levels in SGA newborns. The 11beta-HSD1 expression was similar among the SGA, AGA, and LGA placentas. However, within the placentas of SGA newborns, the 11beta-HSD1 mRNA levels were significantly reduced in the chorionic plate compared with basal plate. An inverse correlation between cord F levels and activity of 11beta-HSD1 in the chorionic plate of the SGA placentas was detected. The 11beta-HSD2 activity was seven- to eightfold higher compared with 11beta-HSD1 in the placentas, and there was a lower 11beta-HSD2 activity in females' SGA placentas compared with the male SGA placentas. CONCLUSION: We observed a lower expression and activity of 11beta-HSD1 in the chorionic plate of the SGA placentas, suggesting a possible compensatory mechanism to diminish the higher cortisol fetal concentrations observed in fetuses with intrauterine growth restriction. PMID- 19542243 TI - Endogenous sex hormones and the prospective association with cardiovascular disease and mortality in men: the Tromso Study. AB - OBJECTIVE: To study the impact of endogenous testosterone levels in community dwelling men on later risk for myocardial infarction (MI) and all-cause, cardiovascular disease (CVD), and ischemic heart disease (IHD) mortality. DESIGN: Population-based prospective cohort study. METHODS: For the analyses, we used a cohort of 1568 randomly selected men, with sex-hormone data participating in the fourth Tromso Study (1994-1995). Defined end points were first-ever MI (fatal or nonfatal), all-cause, CVD, and IHD mortality. A committee performed thorough ascertainment of end points, following a detailed protocol. Complete ascertainment of end points was until 30 September 2007 for all-cause mortality, until 31 December 2005 for CVD/IHD mortality, and until 31 December 2004 for first-ever MI. The prospective association between total and free testosterone and end points were examined using Cox proportional hazard regression, allowing for multivariate adjustment for age and cardiovascular risk factors. RESULTS: During follow-up, there were 395 deaths from all causes, 130 deaths from CVD and 80 deaths from IHD, while 144 men experienced a first-ever MI. There was a significant increase in all-cause mortality risk for men with free testosterone in the lowest quartile (<158 pmol/l) compared with the higher quartiles after age adjustment hazard ratios (HR 1.24, 95% confidence interval, CI 1.01-1.53) and after multivariate adjustments (HR 1.24, 95% CI 1.01-1.54). Total testosterone was not associated with mortality risk. Likewise, there were no significant changes in risk for first-ever MI across different total or free testosterone levels. CONCLUSION: Men with free testosterone levels in the lowest quartile had a 24% increased risk of all-cause mortality. PMID- 19542244 TI - The patient experience of services for thyroid eye disease in the United Kingdom: results of a nationwide survey. AB - BACKGROUND: A recent consensus statement from the European Group on Graves' Orbitopathy recommends referring all patients with thyroid eye disease (TED), except the mildest cases, to a specialist multidisciplinary clinic. OBJECTIVE: To study the patients' experiences of accessing services for the treatment of TED in the UK. METHODS: A postal questionnaire survey of 395 members of two patients support organisations for TED in the UK, the TED Charitable Trust and the British Thyroid Foundation. RESULTS: The response rate was 67%. The majority of responders were females (91%) and aged above 45 (74%). There were delays in the diagnosis and referral. In 26% of responders, the time lapsed from the first symptoms to the diagnosis of TED for over 12 months. There was a wide variation in the type of clinic and healthcare professionals involved in the treatment of TED. Only 25% of the responders attended a specialist TED clinic. Out of these, 33% waited over 6 months from the first consultation with a doctor to being seen at a specialist TED clinic. Only 56% of responders were satisfied with the treatment they received for TED. More responders who had attended a specialist TED clinic were satisfied with the treatment than those who had not attended a specialist clinic (67 vs 52%, P<0.05). CONCLUSION: Only a minority of patients with TED are treated at a specialist TED clinic in the UK. Those patients who are treated at a specialist TED clinic are more likely to be satisfied with the treatment. PMID- 19542246 TI - Mepacrine inhibits subepithelial fibrosis by reducing the expression of arginase and TGF-beta1 in an extended subacute mouse model of allergic asthma. AB - Asthma is a dynamic disorder of airway inflammation and airway remodeling with an imbalance in T helper type 1 (Th(1))/Th(2) immune response. Increased Th(2) cytokines such as IL-4 and IL-13 induce arginase either directly or indirectly through transforming growth factor-beta(1) (TGF-beta(1)) and lead to subepithelial fibrosis, which is a crucial component of airway remodeling. Synthetic antimalarials have been reported to have immunomodulatory properties. Mepacrine is known for its reduction of airway inflammation in short-term allergen challenge model by reducing Th(2) cytokines and cysteinyl leukotrienes, which has an important role in the development of airway remodeling features. Therefore, we hypothesized that mepacrine may reduce airway remodeling. For this, extended subacute ovalbumin mice model of asthma was developed; these mice showed an increased expression of profibrotic mediators, subepithelial fibrosis, and goblet cell metaplasia along with airway inflammation, increased Th(2) cytokines, allergen-specific IgE, IgG(1), increased cytosolic PLA(2) (cPLA(2)), and airway hyperresponsiveness. Presence of intraepithelial eosinophils and significant TGF beta(1) expression in subepithelial mesenchymal regions by repeated allergen exposures indicate that asthmatic mice of this study have developed human mimicking as well as late stages of asthma. However, mepacrine treatment decreased Th(2) cytokines and subepithelial fibrosis and alleviated asthma features. These reductions by mepacrine were associated with a decrease in levels and expression of TGF-beta(1) and the reduction in activity, expression of arginase in lung cytosol, and immunolocalization in inflammatory cells present in perivascular and peribronchial regions. These results suggest that mepacrine might reduce the development of subepithelial fibrosis by reducing the arginase and TGF-beta(1). These effects of mepacrine likely underlie its antiairway remodeling action in asthma. PMID- 19542247 TI - TLR3 activation stimulates cytokine secretion without altering agonist-induced human small airway contraction or relaxation. AB - Respiratory infections exacerbate chronic lung diseases promoting airway inflammation and hyperreactivity. Toll-like receptor 3 (TLR3) recognizes viral double-stranded (ds) RNA such as polyinosinic-polycytidylic acid [poly(I:C)] and stimulates innate immune responses. The objective of this study was to test the hypothesis that dsRNA promotes lung inflammation and alters airway responsiveness to cholinergic and beta-adrenergic receptor agonists in human lung slices. Human airway smooth muscle (ASM) was incubated for 24 h in poly(I:C) +/- TNFalpha and a TLR3 monoclonal antibody. Precision-cut lung slices (PCLS; 250-microm thickness) from healthy human lungs containing a small airway were incubated in 0, 10, or 100 microg/ml poly(I:C) for 24 h. Intravital microscopy of lung slices was used to quantify contractile and relaxation responsiveness to carbachol and isoproterenol, respectively. Supernatants of ASM and PCLS were analyzed for cytokine secretion using a 25-multiplex bead assay. In human ASM, poly(I:C) (0.5 microg/ml) increased macrophage inflammatory protein-1alpha (MIP-1alpha) and RANTES that was prevented by a TLR3 monoclonal receptor antibody. Incubation of human PCLS with poly(I:C) (10 and 100 microg/ml) had little effect on the log EC(50) or maximum drug effect (E(max)) for contraction and relaxation in response to carbachol and isoproterenol, respectively. The responsiveness of the same human PCLS to poly(I:C) incubation was confirmed by the robust increase in chemokines and cytokines. In separate experiments, incubation of PCLS with IL-13 or TNFalpha (100 ng/ml) increased airway sensitivity to carbachol. Poly(I:C) promotes inflammatory mediator release that was not associated with enhanced bronchoconstriction or attenuated bronchodilation in normal healthy human lung slices. Transduction at the TLR3 initiated by dsRNA stimulates downstream innate immune responses. PMID- 19542249 TI - Increased quantity of tumor-infiltrating FOXP3-positive regulatory T cells is an independent predictor for improved clinical outcome in extranodal NK/T-cell lymphoma. AB - BACKGROUND: Extranodal natural killer/T-cell lymphoma (NKTCL) is a clinically heterogeneous disease with a poor prognosis, requiring risk-stratified management in affected patients. Recently, tumor microenvironment including regulatory T cells (Tregs) has been implicated as a prognostic marker in certain types of lymphoma. PATIENTS AND METHODS: We collected 64 NKTCL cases and numerically quantified the amount of tumor-infiltrating FOXP3-positive Tregs by automated slide scanning and image analysis program after immunohistochemical staining using anti-FOXP3 antibody. RESULTS: Patients were able to be classified into two end groups by their level of Tregs. Twenty-eight (44%) patients had Tregs <50/0.40 mm(2), while 36 (56%) had Tregs > or =50/0.40 mm(2) within the tumor. The decreased number of Tregs (<50/0.40 mm(2)) was more common in patients with poor performance status or in those presented in non-upper aerodigestive tract. However, the level of Tregs was not associated with other prognostic factors, including stage, lactate dehydrogenase level, International Prognostic Index, and NKTCL Prognostic Index. Importantly, patients with increased numbers of Tregs (> or =50/0.40 mm(2)) showed prolonged overall and progression-free survival (P = 0.0005 and P = 0.0079, respectively). The number of FOXP3-positive Tregs was an independent prognostic factor (P = 0.001) by multivariate analysis. CONCLUSION: Increased quantity of tumor-infiltrating Tregs predicted improved clinical outcome in NKTCL patients. PMID- 19542245 TI - UTP regulation of ion transport in alveolar epithelial cells involves distinct mechanisms. AB - UTP is known to regulate alveolar fluid clearance. However, the relative contribution of alveolar type I cells and type II cells to this process is unknown. In this study, we investigated the effects of UTP on ion transport in type I-like cell (AEC I) and type II-like cell (AEC II) monolayers. Luminal treatment of cell monolayers with UTP increased short-circuit current (I(sc)) of AEC II but decreased I(sc) of AEC I. The Cl(-) channel blockers NPPB and DIDS inhibited the UTP-induced changes in I(sc) (DeltaIsc) in both types of cells. Amiloride, an inhibitor of epithelial Na(+) channels (ENaC), abolished the UTP induced DeltaI(sc) in AEC I, but not in AEC II. The general blocker of K(+) channels, BaCl(2), eliminated the UTP-induced DeltaI(sc) in AEC II, but not in AEC I. The intermediate conductance (IK(Ca)) blocker, clofilium, also blocked the UTP effect in AEC II. The signal transduction pathways mediated by UTP were the same in AEC I and AEC II. Furthermore, UTP increased Cl(-) secretion in AEC II and Cl(-) absorption in AEC I. Our results suggest that UTP induces opposite changes in I(sc) in AEC I and AEC II, likely due to the reversed Cl(-) flux and different contributions of ENaC and IK(Ca). Our results further imply a new concept that type II cells contribute to UTP-induced fluid secretion and type I cells contribute to UTP-induced fluid absorption in alveoli. PMID- 19542250 TI - A feasibility study of sequential doublet chemotherapy comprising carboplatin doxorubicin and carboplatin-paclitaxel for advanced endometrial adenocarcinoma and carcinosarcoma. AB - BACKGROUND: Platinum compounds, taxanes and anthracyclines provide the major effective drug classes in the treatment of advanced and recurrent endometrial cancer and carcinosarcoma. PATIENTS AND METHODS: A total of 52 women with advanced or recurrent endometrial cancer and carcinosarcoma were treated with four cycles of carboplatin area under the curve (AUC) 5 and doxorubicin (50 mg/m(2)) for four cycles before or after four cycles of carboplatin AUC5 and paclitaxel (175 mg/m(2)) with each cycle administered at 21-day intervals. RESULTS: Thirty-seven patients (71.2%) completed all planned treatment. Excluding six patients who did not complete treatment for non-drug-related causes, 80.4% completed all planned treatment. Three hundred and seventy-one treatment cycles were administered and 303 (81.7%) occurred on time. Common Toxicity Criteria grade 3/4 haematological toxic effects, particularly neutropenia and thrombocytopenia, were the predominant cause of treatment delays and dose reductions. A low incidence of grade 3 neurotoxicity and no cardiac toxicity were observed. The overall response rates for patients with evaluable disease were 82.1% and 66.7% for endometrial and carcinosarcoma, respectively. At a median follow-up of 21 months, the median progression-free survival for the endometrial adenocarcinoma and carcinosarcoma cohorts were 15.3 and 12.0 months, respectively. CONCLUSION: This regimen is generally well tolerated with encouraging efficacy. PMID- 19542251 TI - Combination of sunitinib, cetuximab and irradiation in an orthotopic head and neck cancer model. AB - BACKGROUND: Recent preclinical and clinical studies indicate beneficial effects from combining radiotherapy with either anti-angiogenic drugs or anti-epidermal growth factor receptor (EGFR)-targeting agent. To investigate the effect of combining these approaches, we evaluated in vivo the antitumor efficacy of the anti-angiogenic compound sunitinib, an oral, multi-targeted tyrosine kinase inhibitor that inhibits among others vascular endothelial growth factor (VEGF) receptors-1, -2 and -3, cetuximab, a mAb targeting the EGFR, and irradiation (RT) given alone and in combination. MATERIALS AND METHODS: Investigations were carried out using a VEGF-secreting human head and neck tumor cell line, CAL33, with a high EGFR content, growing as orthotopic xenografts in nude mice. Three days after tumor cell injection, sunitinib (20 mg/kg, p.o.), cetuximab (1 mg/kg, i.p.), both 5 days/week seven doses, and RT (6 Gy, 3 days/week, four doses) were administered alone and in combination during 9 days. RESULTS: Concomitant administration of drugs produced a marked and significant supra-additive decrease, and the addition of RT completely abolished tumor growth. The drug association markedly reduced tumor cell proliferation (Ki67) and the number of the vessels, but enhanced cell differentiation. CONCLUSION: The efficacy of this combination of sunitinib, cetuximab and RT may be of clinical importance in the management of head and neck cancer patients. PMID- 19542253 TI - FGF2 is crucial for the development of bovine luteal endothelial networks in vitro. AB - The development of the corpus luteum requires angiogenesis, and involves the complex interplay between factors such as vascular endothelial growth factor A (VEGFA), fibroblast growth factor 2 (FGF2) and platelet-derived growth factor (PDGF). However, the relative role of these factors remains to be elucidated. This study used a new physiologically relevant mixed luteal cell culture system to test the hypotheses that: a) FGF2 and VEGFA are critical for bovine luteal angiogenesis; and b) local luteal PDGF signalling stimulates the formation of endothelial networks. Cells were treated with receptor tyrosine kinase inhibitors against VEGFA (SU1498), FGF2 (SU5402) or PDGF (AG1295) activity. After 9 days in culture, endothelial cells were immunostained for von Willebrand factor (VWF) and quantified by image analysis. Highly organised intricate endothelial networks were formed in the presence of exogenous VEGFA and FGF2. The inhibition of FGF2 activity reduced the total area of VWF staining versus controls (>95%; P<0.001). Inhibition of VEGF and PDGF activity reduced the endothelial network formation by more than 60 and 75% respectively (P<0.05). Progesterone production increased in all treatments from day 1 to 7 (P<0.001), and was unaffected by FGF2 or PDGF receptor kinase inhibition (P>0.05), but was reduced by the VEGF receptor inhibitor on days 5 and 7 (P<0.001). In conclusion, this study confirmed that VEGF signalling regulates both bovine luteal angiogenesis and progesterone production. However, FGF2 was crucial for luteal endothelial network formation. Also, for the first time, this study showed that local luteal PDGF activity regulates bovine luteal endothelial network formation in vitro. PMID- 19542252 TI - Ca2+-stores in sperm: their identities and functions. AB - Intracellular Ca2+ stores play a central role in the regulation of cellular [Ca2+](i) and the generation of complex [Ca2+] signals such as oscillations and waves. Ca2+ signalling is of particular significance in sperm cells, where it is a central regulator in many key activities (including capacitation, hyperactivation, chemotaxis and acrosome reaction) yet mature sperm lack endoplasmic reticulum and several other organelles that serve as Ca2+ stores in somatic cells. Here, we review i) the evidence for the expression in sperm of the molecular components (pumps and channels) which are functionally significant in the activity of Ca2+ stores of somatic cells and ii) the evidence for the existence of functional Ca2+ stores in sperm. This evidence supports the existence of at least two storage organelles in mammalian sperm, one in the acrosomal region and another in the region of the sperm neck and midpiece. We then go on to discuss the probable identity of these organelles and their discrete functions: regulation by the acrosome of its own secretion and regulation by membranous organelles at the sperm neck (and possibly by the mitochondria) of flagellar activity and hyperactivation. Finally, we consider the ability of the sperm discretely to control mobilisation of these stores and the functional interaction of stored Ca2+ at the sperm neck/midpiece with CatSper channels in the principal piece in regulation of the activities of mammalian sperm. PMID- 19542254 TI - Qualitative analysis of high-resolution CT scans in severe asthma. AB - BACKGROUND: High-resolution CT (HRCT) scanning is part of the management of severe asthma, but its application varies between centers. We sought to describe the HRCT scan abnormalities of a large severe asthma cohort and to determine the utility of clinical features to direct the use of HRCT scanning in this group of patients. METHODS: Subjects attending our Difficult Asthma Clinic (DAC) between February 2000 and November 2006 (n = 463) were extensively re-characterized and 185 underwent HRCT scan. The HRCT scans were analyzed qualitatively and the interobserver variability was assessed. Using logistic regression we defined clinical parameters that were associated with bronchiectasis (BE) and bronchial wall thickening (BWT) alone or in combination. RESULTS: HRCT scan abnormalities were present in 80% of subjects and often coexisted with BWT (62%), BE (40%), and emphysema (8%). The interobserver agreement for BE (kappa = 0.76) and BWT (kappa = 0.63) was substantial. DAC patients who underwent HRCT scanning compared with those who did not were older, had longer disease duration, had poorer lung function, were receiving higher doses of corticosteroids, and had increased neutrophilic airway inflammation. The sensitivity and specificity of detecting BE clinically were 74% and 45%, respectively. FEV(1)/FVC ratio emerged as an important predictor for both BE and BWT but had poor discriminatory utility for subjects who did not have airway structural changes (FEV(1)/FVC ratio, >or= 75%; sensitivity, 67%; specificity, 65%). CONCLUSION: HRCT scan abnormalities are common in patients with severe asthma. Nonradiologic assessments fail to reliably predict important bronchial wall changes; therefore, CT scan acquisition may be required in all patients with severe asthma. PMID- 19542255 TI - Quantitative sensory testing in the oropharynx: a means of showing nervous lesions in patients with obstructive sleep apnea and snoring. AB - BACKGROUND: It is not fully understood why habitual snoring frequently progresses to obstructive sleep apnea syndrome (OSAS). Vibrations per se may cause peripheral nerve lesions. Therefore, snoring vibrations could cause nervous lesions, leading to impaired reflex activation of dilating muscles at inspiration. In this study, the methodology for quantitative sensory testing in the oropharynx was developed, and the presence of sensory nerve lesions in patients with OSAS and snoring was evaluated. METHODS: Vibration detection thresholds (VDTs) and/or cold detection thresholds (CDTs) were tested at the tonsillar pillars, tongue, lip, and finger in 23 nonsnoring individuals, 13 habitual snorers (apnea-hypopnea index [AHI] < 10), and 31 patients with OSAS (AHI > 20). RESULTS: At tonsillar pillars, there were significant gender differences in both VDT and CDT, with women having lower thresholds. VDT showed no significant differences between any of the three groups when men only were tested. Two nonsnoring control subjects could not detect vibrations at all. When both genders were tested, there was significant difference only between nonsnorers and patients with OSAS (p = 0.003). CDT showed significant differences between nonsnorers and snorers (p = 0.001) and also between nonsnorers and patients with OSAS (p < 0.001), but not between snorers and patients with OSAS. CDT was easier to test than VDT with low variability in nonsnorers. CONCLUSIONS: CDT gave more discriminative results than VDT. Signs of sensory nervous lesions were present in the oropharynx of most patients with OSAS and some snorers, supporting the hypothesis of a progressive oropharyngeal nervous lesion. CDT testing could be a useful clinical method to evaluate the degree of oropharyngeal nervous lesions in patients who snore and in those with OSAS. PMID- 19542256 TI - Prospective evaluation of right ventricular function and functional status 6 months after acute submassive pulmonary embolism: frequency of persistent or subsequent elevation in estimated pulmonary artery pressure. AB - BACKGROUND: No published data have systematically documented pulmonary artery pressure over an intermediate time period after submassive pulmonary embolism (PE). The aim of this work was to document the rate of pulmonary hypertension, as assessed noninvasively by estimated right ventricular systolic pressure (RVSP) of >or= 40 mm Hg 6 months after the diagnosis of submassive PE. METHODS: We enrolled 200 normotensive patients with CT angiography-proven PE and a baseline echocardiogram to estimate RVSP. All patients received therapy with unfractionated heparin initially, but 21 patients later received alteplase in response to circulatory shock or respiratory failure. Patients returned at 6 months for repeat RVSP measurement, and assessments of the New York Heart Association (NYHA) score and 6-min walk distance (6MWD). RESULTS: Six months after receiving a diagnosis, 162 of 180 survivors (90%) returned for follow-up, including 144 patients who had been treated with heparin (heparin-only group) and 18 patients who had been treated with heparin plus alteplase (heparin-plus alteplase group). Among the heparin-only patients, the RVSP at diagnosis was >or= 40 mm Hg in 50 of 144 patients (35%; 95% CI, 27% to 43%), compared with 10 of 144 patients at follow-up (7%; 95% CI, 3% to 12%). However, the RVSP at follow-up was higher than the baseline RVSP in 39 of 144 patients (27%; 95% CI, 9% to 35%), and 18 of these 39 patients had a NYHA score of >or= 3 or exercise intolerance (6MWD, < 330 m). Among heparin-plus-alteplase patients, the RVSP was >or= 40 mm Hg in 11 of 18 patients at diagnosis (61%; 95% CI, 36% to 83%), compared with 2 of 18 patients at follow-up (11%; 95% CI, 1% to 35%). The RVSP at follow-up was not higher than at the time of diagnosis in any of the heparin-plus-alteplase patients (95% CI, 0% to 18%). CONCLUSIONS: Six months after experiencing submassive PE, a significant proportion of patients had echocardiographic and functional evidence of pulmonary hypertension. PMID- 19542257 TI - The metabolic syndrome in patients with chronic bronchitis and COPD: frequency and associated consequences for systemic inflammation and physical inactivity. AB - BACKGROUND: The metabolic syndrome is a condition frequently found among individuals > 60 years of age. It predisposes affected individuals to systemic inflammation and physical inactivity. Systemic inflammation and physical inactivity are relevant extrapulmonary markers of morbidity and mortality in patients with COPD. Here, we studied the following: (1) the frequency of the coexisting metabolic syndrome in patients with chronic bronchitis (CB) and COPD of different severities; and (2) its association with systemic inflammation and physical inactivity. METHODS: In 30 patients with CB (normal spirometry finding) and in 170 patients with COPD (Global Initiative for Chronic Obstructive Lung Disease [GOLD] stages I to IV), we measured the characteristics of the metabolic syndrome, systemic inflammation (high-sensitivity C-reactive protein [hs-CRP], interleukin-6, fibrinogen), and the physical activity level. RESULTS: The frequencies of the metabolic syndrome in patients with CB, GOLD stages I, II, III, and IV, were 53%, 50%, 53%, 37%, and 44%, respectively (average, 47.5%). The levels of hs-CRP and interleukin-6 were significantly increased in patients with the metabolic syndrome, while the physical activity level was significantly decreased. Multivariate linear regression analyses revealed metabolic syndrome, physical activity level, and CB/GOLD stages to be independent predictors of hs CRP and interleukin-6 levels, and physical activity level to be a predictor of fibrinogen levels. CONCLUSIONS: In our study, almost one-half of the patients with CB/COPD had coexisting metabolic syndrome, with a slightly lower frequency in patients with severe COPD. The coexisting metabolic syndrome is associated with an increase in the levels of some systemic inflammatory markers and physical inactivity, independent of lung function impairment. PMID- 19542258 TI - Characteristics of children intubated and mechanically ventilated in 16 PICUs. AB - BACKGROUND: When designing multicenter clinical trials, it is important to understand the characteristics of children who have received ventilation in PICUs. METHODS: This study involved the secondary analysis of an existing data set of all children intubated and mechanically ventilated from 16 US PICUs who were initially screened for a multicenter clinical trial on pediatric acute lung injury (ALI). RESULTS: A total of 12,213 children between 2 weeks and 18 years of age who were intubated and mechanically ventilated were included, representing 30% of PICU admissions (center range, 20 to 64%). Of the children who received ventilation, 22% had cyanotic congenital heart disease; 26% had respiratory failure but not bilateral pulmonary infiltrates on chest radiograph; 8% had chronic respiratory disease; 7% had upper airway obstruction; and 5% had reactive airway disease. At least 1,457 patients (15%) with respiratory failure lacked an arterial line. Of these patients, 97% had a positive end-expiratory pressure 18 months of chemotherapy. CONCLUSIONS: The higher dose of CAM allowed for better culture conversion. Daily combination chemotherapy that includes CAM (800 mg) seems appropriate as an initial treatment against treatment naive patients with nodular and bronchiectatic MAC pulmonary disease. PMID- 19542260 TI - Effects of sleep fragmentation on glucose metabolism in normal subjects. AB - BACKGROUND: Sleep disorders are increasingly associated with insulin resistance, glucose intolerance, and type 2 diabetes mellitus. Whether the metabolic toll imposed by sleep-related disorders is caused by poor-quality sleep or due to other confounding factors is not known. The objective of this study was to examine whether experimental sleep fragmentation across all sleep stages would alter glucose metabolism, adrenocortical function, and sympathovagal balance. METHODS: Sleep was experimentally fragmented across all stages in 11 healthy, normal volunteers for two nights using auditory and mechanical stimuli. Primary outcomes included insulin sensitivity (S(I)), glucose effectiveness (S(G)), and insulin secretion, as determined by the intravenous glucose tolerance test. Secondary outcomes included measures of sympathovagal balance and serum levels of inflammatory markers, adipokines, and cortisol. RESULTS: Following two nights of sleep fragmentation, S(I) decreased from 5.02 to 3.76 (mU/L)(-1)min(-1) (P < .0001). S(G), which is the ability of glucose to mobilize itself independent of an insulin response, also decreased from 2.73 x 10(-2) min(-1) to 2.16 x 10(-2) min(-1) (P < .01). Sleep fragmentation led to an increase in morning cortisol levels and a shift in sympathovagal balance toward an increase in sympathetic nervous system activity. Markers of systemic inflammation and serum adipokines were unchanged with sleep fragmentation. CONCLUSIONS: Fragmentation of sleep across all stages is associated with a decrease in S(I) and S(G). Increases in sympathetic nervous system and adrenocortical activity likely mediate the adverse metabolic effects of poor sleep quality. PMID- 19542261 TI - Standards for sample composition and impairment classification in neuropsychological studies of multiple sclerosis. PMID- 19542263 TI - Measures in the first year of therapy predict the response to interferon beta in MS. AB - BACKGROUND AND OBJECTIVE: Several criteria for treatment response to interferon beta (IFNbeta) have been proposed, although there is no consensus among different investigators. Hence, the aim of this study was to investigate magnetic resonance imaging (MRI) and clinical predictors of response during the first 12 months of therapy. METHODS: This is a prospective and longitudinal study of relapsing remitting multiple sclerosis (RRMS) patients treated with IFNbeta. Patients were classified based on the presence of new lesions on MRI, relapses, confirmed disability increase, or combinations of all these variables after 1 year of therapy. Regression analysis was performed in order to identify variables of response after a follow-up of 3 years. RESULTS: We included 222 RRMS patients. The logistic model demonstrated that only the combination of new active lesions on MRI with the presence of relapses (OR 4.4; 95% CI 1.6-12.5) or disability progression (Odds Ratio (OR) 7.1; 95% Confidence Interval (CI) 1.6-33.9), or both (OR 6.5; 95% CI 1.9-23.4) achieved significant values to identify those patients with a poor outcome. CONCLUSIONS: In RRMS patients treated with IFNbeta, the combination of measures of disease activity and the presence of new active lesions on MRI may have a prognostic value for identifying patients with disease activity in the second and third year of therapy. PMID- 19542264 TI - Cognitive deficits in multiple sclerosis patients with cerebellar symptoms. AB - BACKGROUND: Cerebellar dysfunction is common in patients with multiple sclerosis (MS). However, neuropsychological studies of this clinical feature are lacking. OBJECTIVE: We investigate the neuropsychological features in relapsing-remitting MS (RR-MS) patients with and without cerebellar dysfunction. METHODS: Twenty-one RR-MS patients with cerebellar dysfunction (RR-MSc), characterized by prevalent ataxic gait and nystagmus, and 21 RR-MS patients without any cerebellar manifestation (RR-MSnc) pair-matched for demographical and clinical variables were studied. All patients from each group underwent an extensive battery of neuropsychological tests. Magnetic resonance imaging analysis included hyperintense fast fluid-attenuated inversion-recovery lesion load in the whole brain as well as in the four lobes separately. RESULTS: Any significant differences were detected in total and regional lesion load measurements between the two groups. RR-MSc group performed equally as well as the RR-MSnc group on many of the cognitive exploration measures. Nevertheless, the RR-MSc group performed more poorly than the RR-MSnc group on attention tests (Symbol Digit Modalities Test) and verbal fluency tests (Controlled Oral Word Association Test); neither of the test results proved to be affected by regional lesion loads. CONCLUSION: These results highlight the importance of considering cognitive deficits associated with the presence of cerebellar symptoms in RR-MS. PMID- 19542265 TI - Multiple sclerosis produces significant changes in urinary bladder innervation which are partially reflected in the lower urinary tract functional status sensory nerve fibers role in detrusor overactivity. AB - BACKGROUND: Detrusor overactivity is often observed in patients with multiple sclerosis (MS), and neurotoxins are emerging as second-line therapies albeit with different degrees of success per patient basis. OBJECTIVE: To investigate lower urinary tract (LUT) functional status and bladder innervation (calcitonin gene related peptide [CGRP] and substance P [SP] positive nerve fibers) in patients with MS. METHOD: Eighteen MS patients with LUT symptoms underwent urodynamic investigations, and six non-MS patients undergoing cystoscopy due to microscopic hematuria served as controls. Cold cut bladder biopsies were taken from the bladder trigone region. Neurotransmitter expression was determined by individual immunohistochemical staining. RESULTS: Two distinct groups could be distinguished: group 1 with pronounced neurogenic detrusor overactivity and mild outflow obstruction; group 2 with some degree of neurogenic detrusor overactivity, detrusor hypocontractility during voiding, and high degree of an outflow obstruction. The presence of SP and CGRP immunoreactive + fiber density was observed in greater numbers in group 1. CONCLUSION: Density of CGRP and SP positive nerve fibers within the urinary bladder of patients with MS may be suggestive of functional status of the lower urinary tract, namely denser innervation is observed in patients with mild outflow obstruction and strong detrusor overactivity. This observation could be useful when planning second-line treatment (neurotoxins) in these patients. Patients with denser innervation probably will respond better to such a therapy. PMID- 19542262 TI - Cognitive impairment and its relation with disease measures in mildly disabled patients with relapsing-remitting multiple sclerosis: baseline results from the Cognitive Impairment in Multiple Sclerosis (COGIMUS) study. AB - BACKGROUND: Cognitive impairment is a common symptom of multiple sclerosis (MS), but the association between cognitive impairment and magnetic resonance imaging (MRI) disease measures in patients with relapsing-remitting (RR) MS is unclear. OBJECTIVES: To study the prevalence of cognitive impairment and its relation with MRI disease measures in mildly disabled patients with RRMS. METHODS: Patients aged 18-50 years with RRMS (McDonald criteria) and an Expanded Disability Status Scale (EDSS) score or=3 cognitive tests) was present in approximately 20% of all patients and in the subgroup who underwent MRI. T2 hyperintense and T1 hypointense lesion volumes were significantly higher in patients with cognitive impairment (defined as impaired performance on at least three tests of the Rao's battery) than those without. EDSS score was also significantly higher in cognitively impaired than in cognitively preserved patients. Disease duration, depression, and years in formal education did not differ significantly between cognitively impaired and cognitively preserved patients. T2 lesion volume, performance intelligence quotient, and age were significant predictors of cognitive impairment in this population. Weak correlations were found between performance on individual cognitive tests and specific MRI measures, with T1 and T2 lesion volumes correlating with performance on most cognitive tests. CONCLUSIONS: Cognitive impairment occurs in approximately one-fifth of mildly disabled patients with MS and is associated with specific MRI disease measures. Assessment of cognitive function at diagnosis could facilitate the identification of patients who may benefit from therapeutic intervention with disease-modifying therapies to prevent further lesion development. PMID- 19542266 TI - Proposed modifications to the McDonald criteria for use in Asia. PMID- 19542267 TI - Geographical variation in the provision of elective primary hip and knee replacement: the role of socio-demographic, hospital and distance variables. AB - BACKGROUND: To explore inequalities in the provision of hip/knee replacement surgery and produce small-area estimates of provision to inform local health planning. METHODS: Hospital Episode Statistics were used to explore inequalities in the provision of primary hip/knee operations in English NHS hospitals in 2002. Multilevel Poisson regression modelling was used to estimate rates of surgical provision by socio-demographic, hospital and distance variables. GIS software was used to estimate road travel times and create hospital catchment areas. RESULTS: Rates of joint replacement increased with age before falling in those aged 80+. Women received more operations than men. People living in the most deprived areas obtained fewer hip, but more knee operations. Those in urban areas received less hip surgery, but there was no association for knee replacement. Controlling for hospital and distance measures did not attenuate the effects. Geographical variation across districts was observed with some districts showing inequality in socio-demographic factors, whereas others showed none at all. CONCLUSIONS: This study found evidence of inequalities in the provision of joint replacement surgery. However, before we can conclude that there is inequity in receipts of healthcare, future research must consider whether these patterns are explained by variations in need across socio-demographic groups. PMID- 19542268 TI - Reaching every district (RED) approach to strengthen routine immunization services: evaluation in the African region, 2005. AB - BACKGROUND: This evaluation was undertaken in 2005, in the African region, to better understand the reaching every district (RED) implementation process that provides a framework for strengthening immunization services at the district level. METHODS: In June 2005, a convenience sample of five countries was selected to evaluate the implementation of RED. Evaluation teams consisting of key partners conducted site visits to the national, district and health facility levels using standardized qualitative questionnaires. RESULTS: RED was implemented in a similar manner in all five countries, i.e. starting with training and micro-planning. All RED components were implemented to some degree in the countries. Common implementation factors included development of plans, expanding outreach services (defined as services provided in sites outside fixed immunization sites), planning of supervisory visits and efforts to link with communities and utilize community volunteers. Monitoring tools such as wall charts and maps were observed and reportedly used. CONCLUSIONS: Evaluation of the RED implementation process provided evidence of improvement in delivery of routine immunization services. The RED framework should continue to be used to strengthen the immunization delivery system to meet continuing new demands, such as the introduction of new vaccines and integrated delivery of other child survival interventions. PMID- 19542269 TI - High rate of transmission among contacts in large London outbreak of isoniazid mono-resistant tuberculosis. AB - BACKGROUND: For a continuing London outbreak of isoniazid mono-resistant tuberculosis (TB), we aimed to determine transmission rates and risk factors for contacts of early cases, in order to inform future guidance on contact tracing. METHODS: Paper-based proformas were completed by TB nurses, and then analysed using EpiInfo/SAS statistical software. RESULTS: Forty community contacts (11%) became cases, 45 (13%) were recommended chemoprophylaxis and 270 (76%) were discharged clear of infection. The highest transmission rate was among contacts exposed to two or more cases (29% became cases) and close contacts of sputum smear-positive cases (22%). Other risk factors were being male and exposure to drug-using cases or cases with prison links. The number needed to be screened (NNS) to detect one case was lowest [5 (95% CI: 4-8)] for contacts of sputum smear-positive pulmonary cases, although the NNS was still only 20 (95% CI:8-72) for casual contacts of smear-positive cases. CONCLUSIONS: Transmission of disease to contacts was high (11%) compared with other documented outbreaks (0.7-2%). The results support recommended guidelines for contact tracing but also provide grounds to recommend, for outbreak cases, screening of casual contacts of smear positive cases and contacts exposed to more than one case, drug users or prisoners. PMID- 19542270 TI - Role of the sigmaD-dependent autolysins in Bacillus subtilis population heterogeneity. AB - Exponentially growing populations of Bacillus subtilis contain two morphologically and functionally distinct cell types: motile individuals and nonmotile multicellular chains. Motility differentiation arises because RNA polymerase and the alternative sigma factor sigma(D) activate expression of flagellin in a subpopulation of cells. Here we demonstrate that the peptidoglycan remodeling autolysins under sigma(D) control, LytC, LytD, and LytF, are expressed in the same subpopulation of cells that complete flagellar synthesis. Morphological heterogeneity is explained by the expression of LytF that is necessary and sufficient for cell separation. Moreover, LytC is required for motility but not at the level of cell separation or flagellum biosynthesis. Rather, LytC appears to be important for flagellar function, and motility was restored to a LytC mutant by mutation of either lonA, encoding the LonA protease, or a gene encoding a previously unannotated swarming motility inhibitor, SmiA. We conclude that heterogeneous activation of sigma(D)-dependent gene expression is sufficient to explain both the morphological heterogeneity and functional heterogeneity present in vegetative B. subtilis populations. PMID- 19542271 TI - Polyamines are not required for aerobic growth of Escherichia coli: preparation of a strain with deletions in all of the genes for polyamine biosynthesis. AB - A strain of Escherichia coli was constructed in which all of the genes involved in polyamine biosynthesis--speA (arginine decarboxylase), speB (agmatine ureohydrolase), speC (ornithine decarboxylase), spe D (adenosylmethionine decarboxylase), speE (spermidine synthase), speF (inducible ornithine decarboxylase), cadA (lysine decarboxylase), and ldcC (lysine decarboxylase)--had been deleted. Despite the complete absence of all of the polyamines, the strain grew indefinitely in air in amine-free medium, albeit at a slightly (ca. 40 to 50%) reduced growth rate. Even though this strain grew well in the absence of the amines in air, it was still sensitive to oxygen stress in the absence of added spermidine. In contrast to the ability to grow in air in the absence of polyamines, this strain, surprisingly, showed a requirement for polyamines for growth under strictly anaerobic conditions. PMID- 19542272 TI - Optical mapping reveals a large genetic inversion between two methicillin resistant Staphylococcus aureus strains. AB - Staphylococcus aureus is a highly versatile and evolving bacterium of great clinical importance. S. aureus can evolve by acquiring single nucleotide polymorphisms and mobile genetic elements and by recombination events. Identification and location of novel genomic elements in a bacterial genome are not straightforward, unless the whole genome is sequenced. Optical mapping is a new tool that creates a high-resolution, in situ ordered restriction map of a bacterial genome. These maps can be used to determine genomic organization and perform comparative genomics to identify genomic rearrangements, such as insertions, deletions, duplications, and inversions, compared to an in silico (virtual) restriction map of a known genome sequence. Using this technology, we report here the identification, approximate location, and characterization of a genetic inversion of approximately 500 kb of a DNA element between the NRS387 (USA800) and FPR3757 (USA300) strains. The presence of the inversion and location of its junction sites were confirmed by site-specific PCR and sequencing. At both the left and right junction sites in NRS387, an IS1181 element and a 73-bp sequence were identified as inverted repeats, which could explain the possible mechanism of the inversion event. PMID- 19542273 TI - Genome sequence of the emerging pathogen Helicobacter canadensis. AB - We determined the genome sequence of the type strain of Helicobacter canadensis, an emerging human pathogen with diverse animal reservoirs. Potential virulence determinants carried by the genome include systems for N-linked glycosylation and capsular export. A protein-based phylogenetic analysis places H. canadensis close to Wolinella succinogenes. PMID- 19542274 TI - CodY activates transcription of a small RNA in Bacillus subtilis. AB - Regulatory small RNAs (sRNAs) in bacterial genomes have become a focus of research over the past 8 years. Whereas more than 100 such sRNAs have been found in Escherichia coli, relatively little is known about sRNAs in gram-positive bacteria. Using a computational approach, we identified two sRNAs in intergenic regions of the Bacillus subtilis genome, SR1 and SR2 (renamed BsrF). Recently, we demonstrated that SR1 inhibits the translation initiation of the transcriptional activator AhrC. Here, we describe detection of BsrF, its expression profile, and its regulation by CodY. Furthermore, we mapped the secondary structure of BsrF. BsrF is expressed in complex and minimal media in all growth phases in B. subtilis and, with a similar expression profile, also in Bacillus amyloliquefaciens. Neither overexpression nor deletion of bsrF affected the growth of B. subtilis. BsrF was found to be long-lived in complex and minimal media. Analysis of 13 putative transcription factor binding sites upstream of bsrF revealed only an effect for CodY. Here, we showed by using Northern blotting, lacZ reporter gene fusions, in vitro transcription, and DNase I footprinting that the transcription of bsrF is activated by CodY in the presence of branched-chain amino acids and GTP. Furthermore, BsrF transcription was increased 1.5- to 2-fold by glucose in the presence of branched-chain amino acids, and this increase was independent of the known glucose-dependent regulators. BsrF is the second target for which transcriptional activation by CodY has been discovered. PMID- 19542275 TI - CtrA, a global response regulator, uses a distinct second category of weak DNA binding sites for cell cycle transcription control in Caulobacter crescentus. AB - CtrA controls cell cycle programs of chromosome replication and genetic transcription. Phosphorylated CtrA approximately P exhibits high affinity (dissociation constant [K(d)], <10 nM) for consensus TTAA-N7-TTAA binding sites with "typical" (N = 7) spacing. We show here that ctrA promoters P1 and P2 use low-affinity (K(d), >500 nM) CtrA binding sites with "atypical" (N not equal 7) spacing. Footprints demonstrated that phosphorylated CtrA approximately P does not exhibit increased affinity for "atypical" sites, as it does for sites in the replication origin. Instead, high levels of CtrA (>10 microM) accumulate, which can drive CtrA binding to "atypical" sites. In vivo cross-linking showed that when the stable CtrADelta3 protein persists during the cell cycle, the "atypical" sites at ctrA and motB are persistently bound. Interestingly, the cell cycle timing of ctrA P1 and P2 transcription is not altered by persistent CtrADelta3 binding. Therefore, operator DNA occupancy is not sufficient for regulation, and it is the cell cycle variation of CtrA approximately P phosphorylation that provides the dominant "activation" signal. Protein dimerization is one potential means of "activation." The glutathione S-transferase (GST) protein dimerizes, and fusion with CtrA (GST-CtrA) creates a stable dimer with enhanced affinity for TTAA motifs. Electrophoretic mobility shift assays with GST-CtrA revealed cooperative modes of binding that further distinguish the "atypical" sites. GST CtrA also binds a single TTAA motif in ctrA P1 aided by DNA in the extended TTAACCAT motif. We discuss how "atypical" sites are a common yet distinct category of CtrA regulatory sites and new implications for the working and evolution of cell cycle control networks. PMID- 19542276 TI - Cell envelope perturbation induces oxidative stress and changes in iron homeostasis in Vibrio cholerae. AB - The Vibrio cholerae type II secretion (T2S) machinery is a multiprotein complex that spans the cell envelope. When the T2S system is inactivated, cholera toxin and other exoproteins accumulate in the periplasmic compartment. Additionally, loss of secretion via the T2S system leads to a reduced growth rate, compromised outer membrane integrity, and induction of the extracytoplasmic stress factor RpoE (A. E. Sikora, S. R. Lybarger, and M. Sandkvist, J. Bacteriol. 189:8484 8495, 2007). In this study, gene expression profiling reveals that inactivation of the T2S system alters the expression of genes encoding cell envelope components and proteins involved in central metabolism, chemotaxis, motility, oxidative stress, and iron storage and acquisition. Consistent with the gene expression data, molecular and biochemical analyses indicate that the T2S mutants suffer from internal oxidative stress and increased levels of intracellular ferrous iron. By using a tolA mutant of V. cholerae that shares a similar compromised membrane phenotype but maintains a functional T2S machinery, we show that the formation of radical oxygen species, induction of oxidative stress, and changes in iron physiology are likely general responses to cell envelope damage and are not unique to T2S mutants. Finally, we demonstrate that disruption of the V. cholerae cell envelope by chemical treatment with polymyxin B similarly results in induction of the RpoE-mediated stress response, increased sensitivity to oxidants, and a change in iron metabolism. We propose that many types of extracytoplasmic stresses, caused either by genetic alterations of outer membrane constituents or by chemical or physical damage to the cell envelope, induce common signaling pathways that ultimately lead to internal oxidative stress and misregulation of iron homeostasis. PMID- 19542277 TI - CsrRS regulates group B Streptococcus virulence gene expression in response to environmental pH: a new perspective on vaccine development. AB - To identify factors involved in the response of group B streptococci (GBS) to environmental pH, we performed a comparative global gene expression analysis of GBS at acidic and neutral pHs. We found that the transcription of 317 genes was increased at pH 5.5 relative to that at pH 7.0, while 61 genes were downregulated. The global response to acid stress included the differential expression of genes involved in transport, metabolism, stress response, and virulence. Known vaccine candidates, such as BibA and pilus components, were also regulated by pH. We observed that many of the genes involved in the GBS response to pH are known to be controlled by the CsrRS two-component system. Comparison of the regulon of wild-type strain 2603 V/R with that of a csrRS deletion mutant strain revealed that the pH-dependent regulation of 90% of the downregulated genes and 59.3% of the up-regulated genes in strain 2603 V/R was CsrRS dependent and that many virulence factors were overexpressed at high pH. Beta-hemolysin regulation was abrogated by selective inactivation of csrS, suggesting the implication of the CsrS protein in pH sensing. These results imply that the translocation of GBS from the acidic milieu of the vagina to the neutral pH of the neonatal lung signals the up-regulation of GBS virulence factors and conversion from a colonizing to an invasive phenotype. In addition, the fact that increased exposure of BibA on the bacterial surface at pH 7.0 induced opsonophagocytic killing of GBS in immune serum highlights the importance of pH regulation in the protective efficacy of specific antibodies to surface-exposed GBS proteins. PMID- 19542278 TI - Enhancement of the synthesis of RpoE and StpA by polyamines at the level of translation in escherichia coli under heat shock conditions. AB - Proteins whose synthesis is enhanced by polyamines at the level of translation were identified with a polyamine-requiring mutant cultured in the presence of 0.1% glucose and 0.02% glutamate at 42 degrees C. Polyamines had a greater effect on cell growth at 42 degrees C than at 37 degrees C. At 42 degrees C, the synthesis of RpoE (sigma(24)) and StpA, which are involved in the transcription of a number of heat shock response genes, was stimulated by polyamines at the level of translation. In the rpoE and stpA mRNAs, a Shine-Dalgarno (SD) sequence is located at 13 and 12 nucleotides, respectively, upstream of the initiation codon AUG. When the SD sequences were moved to the more common position 7 nucleotides upstream of the initiation codon AUG, the degree of polyamine stimulation was reduced, although the level of RpoE and StpA synthesis was markedly increased. The mechanism underlying polyamine stimulation of RpoE synthesis was then studied. Polyamine stimulation of RpoE synthesis was reduced by changing the bulged-out structure in the initiation site of rpoE mRNA, although the level of RpoE synthesis increased. A selective structural change of this bulged-out region induced by spermidine at 42 degrees C was observed by circular dichroism. Polyamine stimulation of fMet-tRNA binding to ribosomes at 42 degrees C also disappeared by changing the bulged-out structure in the initiation site of rpoE mRNA. The results suggest that polyamines enhance the synthesis of RpoE by changing the bulged-out structure in the initiation site of rpoE mRNA. PMID- 19542279 TI - Proteomic and genomic characterization of highly infectious Clostridium difficile 630 spores. AB - Clostridium difficile, a major cause of antibiotic-associated diarrhea, produces highly resistant spores that contaminate hospital environments and facilitate efficient disease transmission. We purified C. difficile spores using a novel method and show that they exhibit significant resistance to harsh physical or chemical treatments and are also highly infectious, with <7 environmental spores per cm(2) reproducibly establishing a persistent infection in exposed mice. Mass spectrometric analysis identified approximately 336 spore-associated polypeptides, with a significant proportion linked to translation, sporulation/germination, and protein stabilization/degradation. In addition, proteins from several distinct metabolic pathways associated with energy production were identified. Comparison of the C. difficile spore proteome to those of other clostridial species defined 88 proteins as the clostridial spore "core" and 29 proteins as C. difficile spore specific, including proteins that could contribute to spore-host interactions. Thus, our results provide the first molecular definition of C. difficile spores, opening up new opportunities for the development of diagnostic and therapeutic approaches. PMID- 19542280 TI - Effect of perturbation of ATP level on the activity and regulation of nitrogenase in Rhodospirillum rubrum. AB - Nitrogenase activity in Rhodospirillum rubrum and in some other photosynthetic bacteria is regulated in part by the availability of light. This regulation is through a posttranslational modification system that is itself regulated by P(II) homologs in the cell. P(II) is one of the most broadly distributed regulatory proteins in nature and directly or indirectly senses nitrogen and carbon signals in the cell. However, its possible role in responding to light availability remains unclear. Because P(II) binds ATP, we tested the hypothesis that removal of light would affect P(II) by changing intracellular ATP levels, and this in turn would affect the regulation of nitrogenase activity. This in vivo test involved a variety of different methods for the measurement of ATP, as well as the deliberate perturbation of intracellular ATP levels by chemical and genetic means. To our surprise, we found fairly normal levels of nitrogenase activity and posttranslational regulation of nitrogenase even under conditions of drastically reduced ATP levels. This indicates that low ATP levels have no more than a modest impact on the P(II)-mediated regulation of NifA activity and on the posttranslational regulation of nitrogenase activity. The relatively high nitrogenase activity also shows that the ATP-dependent electron flux from dinitrogenase reductase to dinitrogenase is also surprisingly insensitive to a depleted ATP level. These in vivo results disprove the simple model of ATP as the key energy signal to P(II) under these conditions. We currently suppose that the ratio of ADP/ATP might be the relevant signal, as suggested by a number of recent in vitro analyses. PMID- 19542282 TI - Respiration of Escherichia coli can be fully uncoupled via the nonelectrogenic terminal cytochrome bd-II oxidase. AB - The respiratory chain of Escherichia coli is usually considered a device to conserve energy via the generation of a proton motive force, which subsequently may drive ATP synthesis by the ATP synthetase. It is known that in this system a fixed amount of ATP per oxygen molecule reduced (P/O ratio) is not synthesized due to alternative NADH dehydrogenases and terminal oxidases with different proton pumping stoichiometries. Here we show that P/O ratios can vary much more than previously thought. First, we show that in wild-type E. coli cytochrome bo, cytochrome bd-I, and cytochrome bd-II are the major terminal oxidases; deletion of all of the genes encoding these enzymes results in a fermentative phenotype in the presence of oxygen. Second, we provide evidence that the electron flux through cytochrome bd-II oxidase is significant but does not contribute to the generation of a proton motive force. The kinetics support the view that this system is as an energy-independent system gives the cell metabolic flexibility by uncoupling catabolism from ATP synthesis under non-steady-state conditions. The nonelectrogenic nature of cytochrome bd-II oxidase implies that the respiratory chain can function in a fully uncoupled mode such that ATP synthesis occurs solely by substrate level phosphorylation. As a consequence, the yield with a carbon and energy source can vary five- to sevenfold depending on the electron flux distribution in the respiratory chain. A full understanding and control of this distribution open new avenues for optimization of biotechnological processes. PMID- 19542281 TI - SlyA, a MarR family transcriptional regulator, is essential for virulence in Dickeya dadantii 3937. AB - SlyA, a MarR family transcriptional regulator, controls an assortment of biological functions in several animal-pathogenic bacteria. In order to elucidate the functions of SlyA in the phytopathogen Dickeya dadantii (formerly Erwinia chrysanthemi) 3937, a slyA gene deletion mutant (denoted DeltaslyA) was constructed. The mutant exhibited increased sensitivity to sodium hypochlorite, the cationic antimicrobial peptide polymyxin B, and oxidative stress. The mutant showed reduced production of pectate lyase and exopolysaccharide and an inability to form a pellicle. The mutant lacking a functional slyA gene showed a significantly reduced ability to cause maceration of potato tubers. Accordingly, the mutant exhibited significantly reduced bacterial growth and failed to hyperinduce pectate lyase production in planta. Introduction of a plasmid containing slyA into the DeltaslyA mutant caused all of these phenotypes to recover to wild-type levels. These results suggest that SlyA plays an important role in virulence to plants by positively regulating the expression of multiple pathogenicity-related traits of D. dadantii 3937. PMID- 19542284 TI - A grand view of the flagellar motor. PMID- 19542283 TI - Protein domains and residues involved in the CheZ/CheAS interaction. AB - CheZ localizes to chemoreceptor patches by binding CheA-short (CheA(S)). Residues 70 to 134 of CheZ, constituting the apical loops and part of the dimerization domain, suffice for localization. Replacements of Tyr-118, Ile-119, Leu-123, Arg 124, and Leu-126 of CheA interfere with localization. These residues are exposed in the 'P1 domain of CheA(S). PMID- 19542285 TI - Interspecies signaling between Veillonella atypica and Streptococcus gordonii requires the transcription factor CcpA. AB - Streptococcus gordonii and Veillonella atypica, two early-colonizing members of the dental plaque biofilm, participate in a relationship that results in increased transcription of the S. gordonii gene amyB, encoding an alpha-amylase. We show that the transcription factor CcpA is required for this interspecies interaction. PMID- 19542286 TI - igr Genes and Mycobacterium tuberculosis cholesterol metabolism. AB - Recently, cholesterol was identified as a physiologically important nutrient for Mycobacterium tuberculosis survival in chronically infected mice. However, it remained unclear precisely when cholesterol is available to the bacterium and what additional bacterial functions are required for its metabolism. Here, we show that the igr locus, which we previously found to be essential for intracellular growth and virulence of M. tuberculosis, is required for cholesterol metabolism. While igr-deficient strains grow identically to the wild type in the presence of short- and long-chain fatty acids, the growth of these bacteria is completely inhibited in the presence of cholesterol. Interestingly, this mutant is still able to respire under cholesterol-dependent growth inhibition, suggesting that the bacteria can metabolize other carbon sources during cholesterol toxicity. Consistent with this hypothesis, we found that the growth-inhibitory effect of cholesterol in vitro depends on cholesterol import, as mutation of the mce4 sterol uptake system partially suppresses this effect. In addition, the Delta igr mutant growth defect during the early phase of disease is completely suppressed by mutating mce4, implicating cholesterol intoxication as the primary mechanism of attenuation. We conclude that M. tuberculosis metabolizes cholesterol throughout infection. PMID- 19542287 TI - Phylogenetic ubiquity and shuffling of the bacterial RecBCD and AddAB recombination complexes. AB - RecBCD and AddAB are bacterial enzymes that share similar helicase and nuclease activities and initiate repair of DNA double-strand breaks by homologous recombination. Examination of the phylogenetic distribution of AddAB and RecBCD revealed that one or the other complex is present in most sequenced bacteria. In addition, horizontal gene transfer (HGT) events involving addAB and recBCD appear to be common, with the genes encoding one complex frequently replacing those encoding the other. HGT may also explain the unexpected identification of archaeal addAB genes. More than 85% of addAB and recBCD genes are clustered on the genome, suggesting operon structures. A few organisms, including the Mycobacteria, encode multiple copies of these complexes of either the same or mixed classes. The possibility that the enzymatic activities of the AddAB and RecBCD enzymes promote their horizontal transfer is discussed, and the distribution of AddAB/RecBCD is compared to that of the RecU/RuvC resolvases. Finally, it appears that two sequence motifs, the Walker A box involved in ATP binding and an iron-sulfur-cysteine cluster, are present only in subsets of AddB proteins, suggesting the existence of mechanistically distinct classes of AddB. PMID- 19542288 TI - Site-directed mutagenesis identifies a molecular switch involved in copper sensing by the histidine kinase CinS in Pseudomonas putida KT2440. AB - In the presence of copper, Pseudomonas putida activates transcription of cinAQ via the two-component system CinS-CinR. The CinS-CinR TCS was responsive to 0.5 microM copper and was specifically activated only by copper and silver. Modeling studies of CinS identified a potential copper binding site containing H37 and H147. CinS mutants with H37R and H147R mutations had an almost 10-fold reduced copper-dependent induction of cinAQ compared to the wild type. PMID- 19542289 TI - Isolation and characterization of Xenorhabdus nematophila transposon insertion mutants defective in lipase activity against Tween. AB - We identified Xenorhabdus nematophila transposon mutants with defects in lipase activity. One of the mutations, in yigL, a conserved gene of unknown function, resulted in attenuated virulence against Manduca sexta insects. We discuss possible connections between lipase production, YigL, and specific metabolic pathways. PMID- 19542290 TI - Characterization of the Synechocystis strain PCC 6803 penicillin-binding proteins and cytokinetic proteins FtsQ and FtsW and their network of interactions with ZipN. AB - Because very little is known about cell division in noncylindrical bacteria and cyanobacteria, we investigated 10 putative cytokinetic proteins in the unicellular spherical cyanobacterium Synechocystis strain PCC 6803. Concerning the eight penicillin-binding proteins (PBPs), which define three classes, we found that Synechocystis can survive in the absence of one but not two PBPs of either class A or class C, whereas the unique class B PBP (also termed FtsI) is indispensable. Furthermore, we showed that all three classes of PBPs are required for normal cell size. Similarly, the putative FtsQ and FtsW proteins appeared to be required for viability and normal cell size. We also used a suitable bacterial two-hybrid system to characterize the interaction web among the eight PBPs, FtsQ, and FtsW, as well as ZipN, the crucial FtsZ partner that occurs only in cyanobacteria and plant chloroplasts. We showed that FtsI, FtsQ, and ZipN are self-interacting proteins and that both FtsI and FtsQ interact with class A PBPs, as well as with ZipN. Collectively, these findings indicate that ZipN, in interacting with FtsZ and both FtsI and FtQ, plays a similar role to the Escherichia coli FtsA protein, which is missing in cyanobacteria and chloroplasts. PMID- 19542292 TI - Kinematics of intracellular chlamydiae provide evidence for contact-dependent development. AB - A crucial process of chlamydial development involves differentiation of the replicative reticulate body (RB) into the infectious elementary body (EB). We present experimental evidence to provide support for a contact-dependent hypothesis for explaining the trigger involved in differentiation. We recorded live-imaging of Chlamydia trachomatis-infected McCoy cells at key times during development and tracked the temporospatial trajectories of individual chlamydial particles. We found that movement of the particles is related to development. Early to mid-developmental stages involved slight wobbling of RBs. The average speed of particles increased sharply at 24 h postinfection (after the estimated onset of RB to EB differentiation). We also investigated a penicillin supplemented culture containing EBs, RBs, and aberrantly enlarged, stressed chlamydiae. Near-immobile enlarged particles are consistent with their continued tethering to the chlamydial inclusion membrane (CIM). We found a significantly negative, nonlinear association between speed and size/type of particles, providing further support for the hypothesis that particles become untethered near the onset of RB to EB differentiation. This study establishes the relationship between the motion properties of the chlamydiae and developmental stages, whereby wobbling RBs gradually lose contact with the CIM, and RB detachment from the CIM is coincidental with the onset of late differentiation. PMID- 19542293 TI - Investigating translational repression by microRNAs in Arabidopsis. PMID- 19542291 TI - Genetic analysis of activation of the Vibrio cholerae Cpx pathway. AB - The Cpx two-component system is thought to mediate envelope stress responses in many gram-negative bacteria and has been implicated in the pathogenicity of several enteric pathogens. While cues that activate the Escherichia coli Cpx system have been identified, the nature of the molecular signals that stimulate this pathway is not well understood. Here, we investigated stimuli that trigger this system in Vibrio cholerae, a facultative pathogen that adapts to various niches during its life cycle. In contrast to E. coli, there was no basal activity of the V. cholerae Cpx pathway under standard laboratory conditions. Furthermore, several known stimuli of the E. coli pathway did not induce expression of this system in V. cholerae. There were no defects in intestinal growth in V. cholerae cpx mutants, arguing against the idea that this pathway promotes V. cholerae adaptation to conditions in the mammalian host. We discovered that chloride ions activate the V. cholerae Cpx pathway, raising the possibility that this signal transduction system provides a means for V. cholerae to sense and respond to alterations in salinity. We used a genetic approach to screen for mutants in which the Cpx pathway is activated. We found that mutations in genes whose products are required for periplasmic disulfide bond isomerization result in activation of the Cpx pathway, suggesting that periplasmic accumulation of proteins with aberrant disulfide bonds triggers the V. cholerae Cpx pathway. PMID- 19542294 TI - The tomato Pto kinase uses shared and unique surfaces to recognize divergent avirulence proteins. PMID- 19542295 TI - The plastidic bile acid transporter 5 is required for the biosynthesis of methionine-derived glucosinolates in Arabidopsis thaliana. AB - Aliphatic glucosinolate biosynthesis is highly compartmentalized, requiring import of 2-keto acids or amino acids into chloroplasts for side chain elongation and export of the resulting compounds into the cytosol for conversion into glucosinolate. Aliphatic glucosinolate biosynthesis in Arabidopsis thaliana is regulated by three R2R3-MYB transcription factors, the major player being High Aliphatic Glucosinolate 1 (HAG1/MYB28). Here, we show that BAT5, which belongs to the putative bile acid transporter family, is the only member of this family that is transactivated by HAG1/MYB28, HAG2/MYB76, and HAG3/MYB29. Furthermore, two isopropylmalate isomerases genes, IPMI1 and IPMI2, and the isopropylmalate dehydrogenase gene, IPMDH1, were identified as targets of HAG1/MYB28 and the corresponding proteins localized to plastids, suggesting a role in plastidic chain elongation reactions. The BAT proteins also localized to plastids; however, only mutants defective in BAT5 function contained strongly reduced levels of aliphatic glucosinolates. The bat5 mutant chemotype was rescued by induced overexpression of BAT5. Feeding experiments using 2-keto acids and amino acids of different chain length suggest that BAT5 is a plastidic transporter of (chain elongated) 2-keto acids. Mechanical stimuli and methyl jasmonate transiently induced BAT5 expression in inflorescences and leaves. Thus, BAT5 was identified as the first transporter component of the aliphatic glucosinolate biosynthetic pathway. PMID- 19542296 TI - A role for multiple circadian clock genes in the response to signals that break seed dormancy in Arabidopsis. AB - Plant seeds can sense diverse environmental signals and integrate the information to regulate developmental responses, such as dormancy and germination. The circadian clock confers a growth advantage on plants and uses environmental information for entrainment. Here, we show that normal circadian clock gene function is essential for the response to dormancy-breaking signals in seeds. We show that mutations in the clock genes LATE ELONGATED HYPOCOTYL, CIRCADIAN CLOCK ASSOCIATED1 (CCA1), and GIGANTEA (GI) cause germination defects in response to low temperature, alternating temperatures, and dry after-ripening. We demonstrate that the transcriptional clock is arrested in an evening-like state in dry seeds but rapidly entrains to light/dark cycles in ambient temperatures upon imbibition. Consistent with a role for clock genes in seed dormancy control, CCA1 expression is transcriptionally induced in response to dry after-ripening and that after-ripening affects the amplitude of subsequent transcriptional clock gene oscillations. Control of abscisic acid- and gibberellin-related gene expression in seeds requires normal circadian function, and GI and TIMING OF CAB EXPRESSION1 regulate the response to ABA and GA in seeds. We conclude that circadian clock genes play a key role in the integration of environmental signaling controlling dormancy release in plants. PMID- 19542299 TI - Modeling the hydraulics of root growth in three dimensions with phloem water sources. AB - Primary growth is characterized by cell expansion facilitated by water uptake generating hydrostatic (turgor) pressure to inflate the cell, stretching the rigid cell walls. The multiple source theory of root growth hypothesizes that root growth involves transport of water both from the soil surrounding the growth zone and from the mature tissue higher in the root via phloem and protophloem. Here, protophloem water sources are used as boundary conditions in a classical, three-dimensional model of growth-sustaining water potentials in primary roots. The model predicts small radial gradients in water potential, with a significant longitudinal gradient. The results improve the agreement of theory with empirical studies for water potential in the primary growth zone of roots of maize (Zea mays). A sensitivity analysis quantifies the functional importance of apical phloem differentiation in permitting growth and reveals that the presence of phloem water sources makes the growth-sustaining water relations of the root relatively insensitive to changes in root radius and hydraulic conductivity. Adaptation to drought and other environmental stresses is predicted to involve more apical differentiation of phloem and/or higher phloem delivery rates to the growth zone. PMID- 19542298 TI - The rice aquaporin Lsi1 mediates uptake of methylated arsenic species. AB - Pentavalent methylated arsenic (As) species such as monomethylarsonic acid [MMA(V)] and dimethylarsinic acid [DMA(V)] are used as herbicides or pesticides, and can also be synthesized by soil microorganisms or algae through As methylation. The mechanism of MMA(V) and DMA(V) uptake remains unknown. Recent studies have shown that arsenite is taken up by rice (Oryza sativa) roots through two silicon transporters, Lsi1 (the aquaporin NIP2;1) and Lsi2 (an efflux carrier). Here we investigated whether these two transporters also mediate the uptake of MMA(V) and DMA(V). MMA(V) was partly reduced to trivalent MMA(III) in rice roots, but only MMA(V) was translocated to shoots. DMA(V) was stable in plants. The rice lsi1 mutant lost about 80% and 50% of the uptake capacity for MMA(V) and DMA(V), respectively, compared with the wild-type rice, whereas Lsi2 mutation had little effect. The short-term uptake kinetics of MMA(V) can be described by a Michaelis-Menten plus linear model, with the wild type having 3.5 fold higher V(max) than the lsi1 mutant. The uptake kinetics of DMA(V) were linear with the slope being 2.8-fold higher in the wild type than the lsi1 mutant. Heterologous expression of Lsi1 in Xenopus laevis oocytes significantly increased the uptake of MMA(V) but not DMA(V), possibly because of a very limited uptake of the latter. Uptake of MMA(V) and DMA(V) by wild-type rice was increased as the pH of the medium decreased, consistent with an increasing proportion of the undissociated species. The results demonstrate that Lsi1 mediates the uptake of undissociated methylated As in rice roots. PMID- 19542297 TI - AINTEGUMENTA and AINTEGUMENTA-LIKE6 act redundantly to regulate Arabidopsis floral growth and patterning. AB - An Arabidopsis (Arabidopsis thaliana) flower consists of four types of organs arranged in a stereotypical pattern. This complex floral structure is elaborated from a small number of floral meristem cells partitioned from the shoot apical meristem during reproductive development. The positioning of floral primordia within the periphery of the shoot apical meristem depends on transport of the phytohormone auxin with floral anlagen arising at sites of auxin maxima. An early marker of lateral organ fate is the AP2/ERF-type transcription factor AINTEGUMENTA (ANT), which has been proposed to act downstream of auxin in organogenic growth. Here, I show that the related, AINTEGUMENTA-LIKE6 (AIL6)/PLETHORA3 gene acts redundantly with ANT during flower development. ant ail6 double mutants show defects in floral organ positioning, identity, and growth. These floral defects are correlated with changes in the expression levels and patterns of two floral organ identity genes, APETALA3 and AGAMOUS. ant ail6 flowers also display altered expression of an auxin-responsive reporter, suggesting that auxin accumulation and/or responses are not normal. Furthermore, I show that ANT expression in incipient and young floral primordia depends on auxin transport within the inflorescence meristem. These results show that ANT and AIL6 are important regulators of floral growth and patterning and that they may act downstream of auxin in these processes. PMID- 19542300 TI - Minor viral and host genetic polymorphisms can dramatically impact the biologic outcome of an epitope-specific CD8 T-cell response. AB - Human immunodeficiency virus-1 subtypes A and C differ in the highly conserved Gag-TL9 epitope at a single amino acid position. Similarly, the TL9 presenting human leukocyte antigen (HLA) class I molecules B42 and B81 differ only at 6 amino acid positions. Here, we addressed the influence of such minor viral and host genetic variation on the TL9-specific CD8 T-cell response. The clonotypic characteristics of CD8 T-cell populations elicited by subtype A or subtype C were distinct, and these responses differed substantially with respect to the recognition and selection of TL9 variants. Irrespective of the presenting HLA class I molecule, CD8 T-cell responses elicited by subtype C exhibited largely comparable TL9 variant cross-recognition properties, expressed T-cell receptors that used almost exclusively the TRBV 12-3 gene, and selected for predictable patterns of viral variation within TL9. In contrast, subtype A elicited TL9 specific CD8 T-cell populations with completely different, more diverse TCRBV genes and did not select for viral variants. Moreover, TL9 variant cross recognition properties were extensive in B81(+) subjects but limited in B42(+) subjects. Thus, minor viral and host genetic polymorphisms can dramatically alter the immunologic and virologic outcome of an epitope-specific CD8 T-cell response. PMID- 19542301 TI - c-Myc is a target of RNA-binding motif protein 15 in the regulation of adult hematopoietic stem cell and megakaryocyte development. AB - RNA-binding motif protein 15 (RBM15) is involved in the RBM15-megakaryoblastic leukemia 1 fusion in acute megakaryoblastic leukemia. Although Rbm15 has been reported to be required for B-cell differentiation and to inhibit myeloid and megakaryocytic expansion, it is not clear what the normal functions of Rbm15 are in the regulation of hematopoietic stem cell (HSC) and megakaryocyte development. In this study, we report that Rbm15 may function in part through regulation of expression of the proto-oncogene c-Myc. Similar to c-Myc knockout (c-Myc-KO) mice, long-term (LT) HSCs are significantly increased in Rbm15-KO mice due to an apparent LT-HSC to short-term HSC differentiation defect associated with abnormal HSC-niche interactions caused by increased N-cadherin and beta(1) integrin expression on mutant HSCs. Both serial transplantation and competitive reconstitution capabilities of Rbm15-KO LT-HSCs are greatly compromised. Rbm15-KO and c-Myc-KO mice also share related abnormalities in megakaryocyte development, with mutant progenitors producing increased, abnormally small low-ploidy megakaryocytes. Consistent with a possible functional interplay between Rbm15 and c-Myc, the megakaryocyte increase in Rbm15-KO mice could be partially reversed by ectopic c-Myc. Thus, Rbm15 appears to be required for normal HSC-niche interactions, for the ability of HSCs to contribute normally to adult hematopoiesis, and for normal megakaryocyte development; these effects of Rbm15 on hematopoiesis may be mediated at least in part by c-Myc. PMID- 19542302 TI - NFI-A directs the fate of hematopoietic progenitors to the erythroid or granulocytic lineage and controls beta-globin and G-CSF receptor expression. AB - It is generally conceded that selective combinations of transcription factors determine hematopoietic lineage commitment and differentiation. Here we show that in normal human hematopoiesis the transcription factor nuclear factor I-A (NFI-A) exhibits a marked lineage-specific expression pattern: it is upmodulated in the erythroid (E) lineage while fully suppressed in the granulopoietic (G) series. In unilineage E culture of hematopoietic progenitor cells (HPCs), NFI-A overexpression or knockdown accelerates or blocks erythropoiesis, respectively: notably, NFI-A overexpression restores E differentiation in the presence of low or minimal erythropoietin stimulus. Conversely, NFI-A ectopic expression in unilineage G culture induces a sharp inhibition of granulopoiesis. Finally, in bilineage E + G culture, NFI-A overexpression or suppression drives HPCs into the E or G differentiation pathways, respectively. These NFI-A actions are mediated, at least in part, by a dual and opposite transcriptional action: direct binding and activation or repression of the promoters of the beta-globin and G-CSF receptor gene, respectively. Altogether, these results indicate that, in early hematopoiesis, the NFI-A expression level acts as a novel factor channeling HPCs into either the E or G lineage. PMID- 19542303 TI - Long-term clinical outcome of open meniscal allograft transplantation. AB - BACKGROUND: Meniscal allograft transplantation is an accepted treatment for the symptomatic postmeniscectomized knee in younger patients; however, long-term data are scarce on clinical outcome. HYPOTHESIS: Cryopreserved meniscal allograft transplantations can, in the long term, be a good alternative for the symptomatic postmeniscectomized knee in younger patients. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Sixty-three meniscal allografts (40 lateral and 23 medial) were transplanted with an open procedure in 57 patients. Clinical outcome and failure rate were evaluated at 13.8 +/- 2.8 years. Mean age at time of transplantation was 39.4 +/- 6.9 years. RESULTS: Eight medial and 10 lateral allografts failed (overall failure rate, 29%). Overall Lysholm scores significantly improved from 36 +/- 18 preoperatively to 61 +/- 20 at long-term follow-up. Scores were not significantly different in the following subgroups: medial allografts, female patients, and left treated knees. All subgroups had poor scores at mean follow-up of 13.8 years, except the male patients group, which had a fair score. Short-term Lysholm scores were 79 +/- 19 at 3.1 +/- 1.5 years. All subgroups demonstrated a significant difference between short- and long-term Lysholm scores. There were no significant differences in Lysholm scores between posttransplanted survivors and posttransplanted nonsurvivors who received a total knee arthroplasty. For 2 other measures-the Knee injury and Osteoarthritis Outcome Score and the International Knee Documentation Committee scoring system-significant decline was present between men and women only. CONCLUSION: Long-term follow-up results show that meniscal allograft transplantation is a beneficial procedure. Good improvements in clinical function and pain relief have been shown at short-term follow-up in this population. Despite the deterioration in function scores over time, there is still improvement in level of function at long-term follow-up but not at a high level. As such, meniscal allograft transplantation is a good salvage option for the treatment of degenerative arthritis of the symptomatic post-meniscectomized knee. Meniscal allograft transplantation can be used to postpone total knee arthroplasty in younger patients. PMID- 19542304 TI - Treatment of cartilage defects in the knee using alginate beads containing human mature allogenic chondrocytes. AB - BACKGROUND: The repair of osteochondral lesions is imperfect and transient; chondral lesions do not heal in mature cartilage. Attempts have been made to restore cartilage lesions by filling the defects with a temporary artificial biocompatible matrix. PURPOSE: To determine whether the implantation of alginate beads containing human mature allogenic chondrocytes is feasible and safe for the treatment of symptomatic cartilage defects in the knee. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: A biodegradable, alginate-based, biocompatible scaffold containing human mature allogenic chondrocytes was used for the treatment of chondral and osteochondral lesions in the knee. Twenty-one patients were clinically and prospectively evaluated with use of the Western Ontario and McMaster Universities Osteoarthritis Index and a visual analog scale for pain preoperatively and at 3, 6, 9, 12, 18, and 24 months of follow-up. Of the 21 patients, 13 consented to having a biopsy sample taken for investigative purposes from the area of implantation at 12 months of follow-up, allowing histologic assessment of the repair tissue. RESULTS: A statistically significant clinical improvement became apparent after 6 months, and patients improved during the 24 months of follow-up. Adverse reactions to the alginate/fibrin matrix seeded with the allogenic cartilage cells were not observed. Histologic analysis of the biopsy specimens rated the repair tissue as hyaline-like in 15.3% of the samples, as mixed tissue in 46.2%, as fibrocartilage in 30.8%, and as fibrous in 7.7%. CONCLUSION: The results of this short-term pilot study show that the alginate-based scaffold containing human mature allogenic chondrocytes is feasible and safe for the treatment of symptomatic cartilage defects of the knee. The described technique provides clinical and histologic outcomes that are equal but not superior to those of other cartilage repair techniques. PMID- 19542305 TI - An overview of nested genes in eukaryotic genomes. PMID- 19542306 TI - {beta}-glucanase Eng2 is required for ascus wall endolysis after sporulation in the fission yeast Schizosaccharomyces pombe. AB - Meiosis is the developmental program by which sexually reproducing diploid organisms generate haploid gametes. In yeast, meiosis is followed by spore morphogenesis. When Schizosaccharomyces pombe diploid cells undergo meiosis, they differentiate into asci containing four haploid ascospores that are highly resistant to environmental stress. The formation of the ascospore wall requires the activity of several enzymes involved in the biosynthesis and modification of its components, such as alpha- and beta-glucan synthases. Once the spores are completely mature, the wall of the ascus undergoes an endolytic process that results in the release of ascospores from the ascus, allowing their dispersal into the environment. This process requires the activity of the endo-alpha-1,3 glucanase Agn2. Here, we focus on the characterization of the endo-beta-1,3 glucanase Eng2, which is also required for ascospore release from the ascus. Although Eng2 is present during the mitotic cycle, the protein accumulates after meiosis II. The expression of eng2(+) is required for the efficient release of ascospores, as shown by placing eng2(+) under the control of a repressible promoter. Furthermore, a point mutation that destroys the catalytic activity of the protein results in a phenotype similar to that of the mutant strain. Finally, we demonstrate that exogenous addition of purified Eng2 releases the ascospores from asci generated by an eng2Delta mutant. We propose that Eng2 would act together with Agn2 to completely hydrolyze the ascus wall, thereby assisting in the release of ascospores in S. pombe. PMID- 19542307 TI - Remodeling of global transcription patterns of Cryptococcus neoformans genes mediated by the stress-activated HOG signaling pathways. AB - The ability to sense and adapt to a hostile host environment is a crucial element for virulence of pathogenic fungi, including Cryptococcus neoformans. These cellular responses are evoked by diverse signaling cascades, including the stress activated HOG pathway. Despite previous analysis of central components of the HOG pathway, its downstream signaling network is poorly characterized in C. neoformans. Here we performed comparative transcriptome analysis with HOG signaling mutants to explore stress-regulated genes and their correlation with the HOG pathway in C. neoformans. In this study, we not only provide important insights into remodeling patterns of global gene expression for counteracting external stresses but also elucidate novel characteristics of the HOG pathway in C. neoformans. First, inhibition of the HOG pathway increases expression of ergosterol biosynthesis genes and cellular ergosterol content, conferring a striking synergistic antifungal activity with amphotericin B and providing an excellent opportunity to develop a novel therapeutic method for treatment of cryptococcosis. Second, a number of cadmium-sensitive genes are differentially regulated by the HOG pathway, and their mutation causes resistance to cadmium. Finally, we have discovered novel stress defense and HOG-dependent genes, which encode a sodium/potassium efflux pump, protein kinase, multidrug transporter system, and elements of the ubiquitin-dependent system. PMID- 19542308 TI - Production of extracellular polysaccharides by CAP mutants of Cryptococcus neoformans. AB - The human pathogen Cryptococcus neoformans causes meningoencephalitis. The polysaccharide capsule is one of the main virulence factors and consists of two distinct polysaccharides, glucuronoxylomannan (GXM) and galactoxylomannan (GalXM). How capsular polysaccharides are synthesized, transported, and assembled is largely unknown. Previously, it was shown that mutations in the CAP10, CAP59, CAP60, and CAP64 genes result in an acapsular phenotype. Here, it is shown that these acapsular mutants do secrete GalXM and GXM-like polymers. GXM and GalXM antibodies specifically reacted with whole cells and the growth medium of the wild type and CAP mutants, indicating that the capsule polysaccharides adhere to the cell wall and are shed into the environment. These polysaccharides were purified from the medium, either with or without anion-exchange chromatography. Monosaccharide analysis of polysaccharide fractions by gas-liquid chromatography/mass spectrometry showed that wild-type cells secrete both GalXM and GXM. The CAP mutants, on the other hand, were shown to secrete GalXM and GXM like polymers. Notably, the GalXM polymers were shown to contain glucuronic acid. One-dimensional (1)H nuclear magnetic resonance confirmed that the CAP mutants secrete GalXM and also showed the presence of O-acetylated polymers. This is the first time it is shown that CAP mutants secrete GXM-like polymers in addition to GalXM. The small amount of this GXM-like polymer, 1 to 5% of the total amount of secreted polysaccharides, may explain the acapsular phenotype. PMID- 19542309 TI - Role of Ndt80p in sterol metabolism regulation and azole resistance in Candida albicans. AB - The Ndt80p transcription factor modulates azole tolerance in Candida albicans by controlling the expression of the gene for the drug efflux pump Cdr1p. To date, the contribution of this transcriptional modulator to drug tolerance is not yet well understood. Here, we investigate the role of Ndt80p in mediating fluconazole tolerance by determining its genome-wide occupancy using chromatin immunoprecipitation coupled to high-density tiling arrays. Ndt80p was found to bind a large number of gene promoters with diverse biological functions. Gene ontology analysis of these Ndt80p targets revealed a significant enrichment in gene products related to the cell wall, carbohydrate metabolism, stress responses, hyphal development, multidrug transport, and the cell cycle. Ndt80p was found on the promoters of ergosterol biosynthesis genes, including on the azole target Erg11p. Additionally, expression profiling was used to identify fluconazole-responsive genes that require Ndt80p for their proper expression. We found that Ndt80p is crucial for the expression of numerous fluconazole responsive genes, especially genes involved in ergosterol metabolism. Therefore, by combining genome-wide location and transcriptional profiling, we have characterized the Ndt80p fluconazole-dependent regulon and demonstrated the key role of this global transcriptional regulator in modulating sterol metabolism and drug resistance in C. albicans. PMID- 19542311 TI - Adaptations in the glucose metabolism of procyclic Trypanosoma brucei isolates from tsetse flies and during differentiation of bloodstream forms. AB - Procyclic forms of Trypanosoma brucei isolated from the midguts of infected tsetse flies, or freshly transformed from a strain that is close to field isolates, do not use a complete Krebs cycle. Furthermore, short stumpy bloodstream forms produce acetate and are apparently metabolically preadapted to adequate functioning in the tsetse fly. PMID- 19542312 TI - Functional genomics of adhesion, invasion, and mycelial formation in Schizosaccharomyces pombe. AB - Investigation into the switch between single-celled and filamentous forms of fungi may provide insights into cell polarity, differentiation, and fungal pathogenicity. At the molecular level, much of this investigation has fallen on two closely related budding yeasts, Candida albicans and Saccharomyces cerevisiae. Recently, the much more distant fission yeast Schizosaccharomyces pombe was shown to form invasive filaments after nitrogen limitation (E. Amoah Buahin, N. Bone, and J. Armstrong, Eukaryot. Cell 4:1287-1297, 2005) and this genetically tractable organism provides an alternative system for the study of dimorphic growth. Here we describe a second mode of mycelial formation of S. pombe, on rich media. Screening of an S. pombe haploid deletion library identified 12 genes required for mycelial development which encode potential transcription factors, orthologues of S. cerevisiae Sec14p and Tlg2p, and the formin For3, among others. These were further grouped into two phenotypic classes representing different stages of the process. We show that galactose-dependent cell adhesion and actin assembly are both required for mycelial formation and mutants lacking a range of genes controlling cell polarity all produce mycelia but with radically altered morphology. PMID- 19542310 TI - Msb2 signaling mucin controls activation of Cek1 mitogen-activated protein kinase in Candida albicans. AB - We have characterized the role that the Msb2 protein plays in the fungal pathogen Candida albicans by the use of mutants defective in the putative upstream components of the HOG pathway. Msb2, in cooperation with Sho1, controls the activation of the Cek1 mitogen-activated protein kinase under conditions that damage the cell wall, thus defining Msb2 as a signaling element of this pathway in the fungus. msb2 mutants display altered sensitivity to Congo red, caspofungin, zymolyase, or tunicamycin, indicating that this protein is involved in cell wall biogenesis. Msb2 (as well as Sho1 and Hst7) is involved in the transmission of the signal toward Cek1 mediated by the Cdc42 GTPase, as revealed by the use of activated alleles (Cdc42(G12V)) of this protein. msb2 mutants have a stronger defective invasion phenotype than sho1 mutants when tested on certain solid media that use mannitol or sucrose as a carbon source or under hypoxia. Interestingly, Msb2 contributes to growth under conditions of high osmolarity when both branches of the HOG pathway are altered, as triple ssk1 msb2 sho1 mutants (but not any single or double mutant) are osmosensitive. However, this phenomenon is independent of the presence of Hog1, as Hog1 phosphorylation, Hog1 translocation to the nucleus, and glycerol accumulation are not affected in this mutant following an osmotic shock. These results reveal essential functions in morphogenesis, invasion, cell wall biogenesis, and growth under conditions of high osmolarity for Msb2 in C. albicans and suggest the divergence and specialization of this signaling pathway in filamentous fungi. PMID- 19542313 TI - Special Sm core complex functions in assembly of the U2 small nuclear ribonucleoprotein of Trypanosoma brucei. AB - The processing of polycistronic pre-mRNAs in trypanosomes requires the spliceosomal small ribonucleoprotein complexes (snRNPs) U1, U2, U4/U6, U5, and SL, each of which contains a core of seven Sm proteins. Recently we reported the first evidence for a core variation in spliceosomal snRNPs; specifically, in the trypanosome U2 snRNP, two of the canonical Sm proteins, SmB and SmD3, are replaced by two U2-specific Sm proteins, Sm15K and Sm16.5K. Here we identify the U2-specific, nuclear-localized U2B'' protein from Trypanosoma brucei. U2B'' interacts with a second U2 snRNP protein, U2-40K (U2A'), which in turn contacts the U2-specific Sm16.5K/15K subcomplex. Together they form a high-affinity, U2 specific binding complex. This trypanosome-specific assembly differs from the mammalian system and provides a functional role for the Sm core variation found in the trypanosomal U2 snRNP. PMID- 19542315 TI - Effect of 393T>C polymorphism of GNAS1 gene on dobutamine response in Chinese healthy subjects. AB - The purpose of this study was to characterize the functional consequences of the 393T>C polymorphism of the GNAS1 gene in vivo. PCR-RFLP assays were used to identify GNAS1 and beta 1-adrenoceptor genotypes. The heart rate (HR), blood pressure, left ventricular fractional shortening (LVFS), and left ventricular ejection fraction (LVEF) were determined in different genotypes through a modified dobutamine stress echocardiography protocol. Our results showed that individuals with homozygous or heterozygous C393 had an increased cardiovascular agonistic response to dobutamine, and the increases from baseline in LVFS at the 3 dosage levels of dobutamine were 19.3% +/- 1.0% versus 32.0% +/- 2.9%, 36.7% +/ 3.1% versus 41.3% +/- 4.1%, and 51.7% +/- 3.3% versus 58.7% +/- 2.6% in T393 homozygotes and C393 homozygotes or heterozygotes, respectively (P = .026). Significant differences were also found between these 2 groups with the increases from baseline in LVEF (P = .007) and SBP (P = .048). In addition, there were significant differences in the increases from atopine in LVFS (P = .011), LVEF (P = .004), and SBP (P = .046) between the T393 homozygotes and C393 homozygotes or heterozygotes. The change of LVEF in C393 homozygous was higher than that in T393 homozygous at the dose of 40 microg/kg/min (28.9% +/- 4.0% vs 36.4% +/- 2.1%; 95% CI, 18.8%-38.9%; P = .046). These data suggested that the 393T>C polymorphism of GNAS1 was functionally relevant in vivo. PMID- 19542314 TI - Comparison of semiautomated and fully automated methods for QT measurement during a thorough QT/QTc study: variability and sample size considerations. AB - This study compares the ability of 2 semiautomated methods with a fully automated method for QT measurement to minimize the sample size required to detect a moxifloxacin effect and exclude a placebo effect in a thorough QT/QTc study. The fully automated and 1 of 2 semiautomated methods used a global QT measurement in 12 leads, whereas the other semiautomated method used a tangent method on single lead raw complexes. Mean QTcF intervals were greater when measured on a global QT electrocardiogram than on raw complexes, but the mean magnitudes of DeltaQTcF were similar for all methods. The 3 methods detected a statistically significant increase in QTcF for moxifloxacin compared to placebo and were able to exclude a placebo effect on QTcF in all 62 participants. However, due to a smaller variability, the semiautomated methods allowed these detections with fewer than 20 participants, whereas the fully automated required at least 27 participants. PMID- 19542316 TI - Congenital subglottic cyst in a term neonate. PMID- 19542317 TI - Neurological outcome following isolated 10-12 mm fetal ventriculomegaly. PMID- 19542318 TI - Surfactant replacement after acute massive milk aspiration in a very low birthweight infant. PMID- 19542319 TI - Biochemical and electrophysiological characterization of almorexant, a dual orexin 1 receptor (OX1)/orexin 2 receptor (OX2) antagonist: comparison with selective OX1 and OX2 antagonists. AB - Recent preclinical and clinical research has shown that almorexant promotes sleep in animals and humans without disrupting the sleep architecture. Here, the pharmacology and kinetics of [(3)H]almorexant binding to human orexin 1 receptor (OX(1))- and human orexin 2 receptor (OX(2))-human embryonic kidney 293 membranes were characterized and compared with those of selective OX(1) and OX(2) antagonists, including 1-(5-(2-fluoro-phenyl)-2-methyl-thiazol-4-yl)-1-((S)-2-(5 phenyl-(1,3,4)oxadiazol-2-ylmethyl)-pyrrolidin-1-yl)-methanone (SB-674042), 1 (6,8-difluoro-2-methyl-quinolin-4-yl)-3-(4-dimethylamino-phenyl)-urea (SB 408124), and N-ethyl-2-[(6-methoxy-pyridin-3-yl)-(toluene-2-sulfonyl)-amino]-N pyridin-3-ylmethyl-acetamide (EMPA). The effect of these antagonists was also examined in vitro on the spontaneous activity of rat ventral tegmental area (VTA) dopaminergic neurons. [(3)H]Almorexant bound to a single saturable site on hOX(1) and hOX(2) with high affinity (K(d) of 1.3 and 0.17 nM, respectively). In Schild analyses using the [(3)H]inositol phosphates assay, almorexant acted as a competitive antagonist at hOX(1) and as a noncompetitive-like antagonist at hOX(2). In binding kinetic analyses, [(3)H]almorexant had fast association and dissociation rates at hOX(1), whereas it had a fast association rate and a remarkably slow dissociation rate at hOX(2). In the VTA, orexin-A potentiated the basal firing frequency to 175 +/- 17% of control in approximately half of the neurons tested. In the presence of 1 microM SB-674042 or SB-408124, the effect of orexin-A was only partially antagonized. However, in the presence of 1 microM EMPA or 1 microM almorexant, the effect of orexin-A was completely antagonized. In conclusion, almorexant exhibited a noncompetitive and long-lasting pseudo irreversible mode of antagonism as a result of its very slow rate of dissociation from OX(2). The electrophysiology data suggest that OX(2) might be more important than OX(1) in mediating the effect of orexin-A on slow-firing of VTA dopaminergic neurons. PMID- 19542320 TI - Adenovirus-transduced human butyrylcholinesterase in mouse blood functions as a bioscavenger of chemical warfare nerve agents. AB - Human serum butyrylcholinesterase (Hu BChE) is a promising therapeutic against the toxicity of chemical warfare nerve agents. We have showed previously that recombinant (r) Hu BChE can be expressed at very high levels, 400 to 600 U/ml in mouse blood, by delivering the Hu BChE gene using adenovirus (Ad). Here, we report the biochemical properties of the Ad-expressed full-length and truncated rHu BChE in mouse blood. The molecular sizes of the full-length rHu BChE subunit and its oligomers were similar to those of native Hu BChE, although only a small portion of the full-length rHu BChE subunit underwent assembly into dimers and tetramers. As expected, Ad containing the truncated Hu BChE gene transduced the expression of monomeric rHu BChE only. Compared with 415 U of rHu BChE per milliliter in blood, tissues including liver, lung, heart, brain, kidney, muscle, intestine, diaphragm, salivary gland, and fat expressed <10 U/g of rHu BChE activity. Ad-expressed rHu BChE in mouse blood neutralized soman and O-ethyl S-2 N,N-diisopropylaminoethyl methylphosphonothiolate at rates similar to those of native Hu BChE and rHu BChE expressed in vitro. Because the expression of rHu BChE rapidly decreased 6 days after virus administration, sera were assayed for the presence of anti-Hu BChE antibodies. Anti-Hu BChE antibodies were detected on day 7 and in increased amounts thereafter, which coincided with the loss of Hu BChE expression in sera. In conclusion, the delivery of Hu BChE gene using Ad can be a promising strategy that can provide protection against multiple lethal doses of chemical warfare nerve agents in vivo. PMID- 19542322 TI - Abstracts of the British Maternal and Fetal Medicine Society 13th Annual Conference. June 18-19, 2009. Liverpool, United Kingdom. PMID- 19542321 TI - The aldo-keto reductase Akr1b7 gene is a common transcriptional target of xenobiotic receptors pregnane X receptor and constitutive androstane receptor. AB - Aldo-keto reductase (AKR) family 1, member 7 (AKR1B7), a member of the AKR superfamily, has been suggested to play an important role in the detoxification of lipid peroxidation by-products. The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are xenosensors postulated to alleviate xeno- and endobiotic chemical insults. In this study, we show that the mouse Akr1b7 is a shared transcriptional target of PXR and CAR in the liver and intestine. Treatment of wild-type mice with the PXR agonist pregnenolone-16alpha carbonitrile (PCN) activated Akr1b7 gene expression, whereas the effect was abrogated in PXR(-/-) mice. Similarly, the activation of Akr1b7 gene expression by the CAR agonist 1,4-bis[2-(3,5-dichlorpyridyloxyl)]-benzene, seen in wild-type mice, was abolished in CAR(-/-) mice. The promoter of Akr1b7 gene was activated by PXR and CAR, and this activation was achieved through the binding of PXR retinoid X receptor (RXR) or CAR-RXR heterodimers to direct repeat-4 type nuclear receptor-binding sites found in the Akr1b7 gene promoter. At the functional level, treatment with PCN in wild-type mice, but not PXR(-/-) mice, led to a decreased intestinal accumulation of malondialdehyde, a biomarker of lipid peroxidation. The regulation of Akr1b7 by PXR was independent of the liver X receptor (LXR), another nuclear receptor known to regulate this AKR isoform. Because a major function of Akr1b7 is to detoxify lipid peroxidation, the PXR-, CAR-, and LXR-controlled regulatory network of Akr1b7 may have contributed to alleviate toxicity associated with lipid peroxidation. PMID- 19542323 TI - Genetic diversity and multihost pathogenicity of clinical and environmental strains of Burkholderia cenocepacia. AB - A collection of 54 clinical and agricultural isolates of Burkholderia cenocepacia was analyzed for genetic relatedness by using multilocus sequence typing (MLST), pathogenicity by using onion and nematode infection models, antifungal activity, and the distribution of three marker genes associated with virulence. The majority of clinical isolates were obtained from cystic fibrosis (CF) patients in Michigan, and the agricultural isolates were predominantly from Michigan onion fields. MLST analysis resolved 23 distinct sequence types (STs), 11 of which were novel. Twenty-six of 27 clinical isolates from Michigan were genotyped as ST-40, previously identified as the Midwest B. cenocepacia lineage. In contrast, the 12 agricultural isolates represented eight STs, including ST-122, that were identical to clinical isolates of the PHDC lineage. In general, pathogenicity to onions and the presence of the pehA endopolygalacturonase gene were detected only in one cluster of related strains consisting of agricultural isolates and the PHDC lineage. Surprisingly, these strains were highly pathogenic in the nematode Caenorhabditis elegans infection model, killing nematodes faster than the CF pathogen Pseudomonas aeruginosa PA14 on slow-kill medium. The other strains displayed a wide range of pathogenicity to C. elegans, notably the Midwest clonal lineage which displayed high, moderate, and low virulence. Most strains displayed moderate antifungal activity, although strains with high and low activities were also detected. We conclude that pathogenicity to multiple hosts may be a key factor contributing to the potential of B. cenocepacia to opportunistically infect humans both by increasing the prevalence of the organism in the environment, thereby increasing exposure to vulnerable hosts, and by the selection of virulence factors that function in multiple hosts. PMID- 19542324 TI - Variability of Burkholderia pseudomallei strain sensitivities to chlorine disinfection. AB - Burkholderia pseudomallei is a select agent and the causative agent of melioidosis. Variations in previously reported chlorine and monochloramine concentration time (Ct) values for disinfection of this organism make decisions regarding the appropriate levels of chlorine in water treatment systems difficult. This study identified the variation in Ct values for 2-, 3-, and 4 log(10) reductions of eight environmental and clinical isolates of B. pseudomallei in phosphate-buffered water. The greatest calculated Ct values for a 4-log(10) inactivation were 7.8 mg.min/liter for free available chlorine (FAC) at pH 8 and 5 degrees C and 550 mg.min/liter for monochloramine at pH 8 and 5 degrees C. Ionic strength of test solutions, culture hold times in water, and cell washing were ruled out as sources of the differences in prior observations. Tolerance to FAC was correlated with the relative amount of extracellular material produced by each isolate. Solid-phase cytometry analysis using an esterase-cleaved fluorochrome assay detected a 2-log(10)-higher level of organisms based upon metabolic activity than did culture, which in some cases increased Ct values by fivefold. Despite strain-to-strain variations in Ct values of 17-fold for FAC and 2.5-fold for monochloramine, standard FAC disinfection practices utilized in the United States should disinfect planktonic populations of these B. pseudomallei strains by 4 orders of magnitude in less than 10 min at the tested temperatures and pH levels. PMID- 19542325 TI - Anaerobic respiration of elemental sulfur and thiosulfate by Shewanella oneidensis MR-1 requires psrA, a homolog of the phsA gene of Salmonella enterica serovar typhimurium LT2. AB - Shewanella oneidensis MR-1, a facultatively anaerobic gammaproteobacterium, respires a variety of anaerobic terminal electron acceptors, including the inorganic sulfur compounds sulfite (SO3(2-)), thiosulfate (S2O3(2-)), tetrathionate (S4O6(2-)), and elemental sulfur (S(0)). The molecular mechanism of anaerobic respiration of inorganic sulfur compounds by S. oneidensis, however, is poorly understood. In the present study, we identified a three-gene cluster in the S. oneidensis genome whose translated products displayed 59 to 73% amino acid similarity to the products of phsABC, a gene cluster required for S(0) and S2O3(2 ) respiration by Salmonella enterica serovar Typhimurium LT2. Homologs of phsA (annotated as psrA) were identified in the genomes of Shewanella strains that reduce S(0) and S2O3(2-) yet were missing from the genomes of Shewanella strains unable to reduce these electron acceptors. A new suicide vector was constructed and used to generate a markerless, in-frame deletion of psrA, the gene encoding the putative thiosulfate reductase. The psrA deletion mutant (PSRA1) retained expression of downstream genes psrB and psrC but was unable to respire S(0) or S2O3(2-) as the terminal electron acceptor. Based on these results, we postulate that PsrA functions as the main subunit of the S. oneidensis S2O3(2-) terminal reductase whose end products (sulfide [HS-] or SO3(2-)) participate in an intraspecies sulfur cycle that drives S(0) respiration. PMID- 19542326 TI - Synthetic ultrashort cationic lipopeptides induce systemic plant defense responses against bacterial and fungal pathogens. AB - A new family of synthetic, membrane-active, ultrashort lipopeptides composed of only four amino acids linked to fatty acids was tested for the ability to induce systemic resistance and defense responses in plants. We found that two peptides wherein the third residue is a d-enantiomer (italic), C16-KKKK and C16-KLLK, can induce medium alkalinization of tobacco suspension-cultured cells and expression of defense-related genes in cucumber and Arabidopsis seedlings. Moreover, these compounds can prime systemic induction of antimicrobial compounds in cucumber leaves similarly to the plant-beneficial fungus Trichoderma asperellum T203 and provide systemic protection against the phytopathogens Botrytis cinerea B05, Pseudomonas syringae pv. lachrimans, and P. syringae pv. tomato DC3000. Thus, short cationic lipopeptides are a new category of compounds with potentially high utility in the induction of systemic resistance in plants. PMID- 19542327 TI - Genetic determinants of symptoms on viral DNA satellites. AB - Begomovirus-DNA-beta disease complexes induce different symptom phenotypes in their hosts. To investigate the genetic determinants of the phenotypic differences, Nicotiana spp. and tomato plants were inoculated with infectious clones of Tobacco curly shoot virus (TbCSV)/TbCSV DNA-beta (TbCSB) and Tomato yellow leaf curl China virus (TYLCCNV)/TYLCCNV DNA-beta (TYLCCNB) pseudorecombinants and showed that TYLCCNB induced characteristic vein-thickening and enation symptoms, while TbCSB only slightly exacerbated the leaf-curling symptoms, regardless of the helper virus being used. The roles of DNA-beta encoded betaC1 and a 430-nucleotide fragment containing the A-rich region and the putative betaC1 promoter region of the betaC1 gene (referred to as AP) in symptom development were further investigated by constructing hybrid satellites in which the betaC1 coding region or AP was exchanged between the two satellite molecules. A TYLCCNB hybrid with TbCSB betaC1 lost the ability to elicit the vein-thickening and enation phenotypes. TbCSB hybrids containing the TYLCCNB betaC1 or AP fragment failed to induce the characteristic vein thickening and enations. A TYLCCNB hybrid having the TbCSB AP fragment produced the enations, but the number of enations was less and their sizes were reduced. Differently from the phloem specific pattern of the TYLCCNB promoter, a full-length fragment upstream of the TbCSB betaC1 gene confers a constitutive beta-glucuronidase expression pattern in transgenic tobacco plants. The above results indicate that the DNA-beta-encoded betaC1 protein is the symptom determinant, but the promoter of the betaC1 gene has influence on symptom production. PMID- 19542328 TI - Roles of small, acid-soluble spore proteins and core water content in survival of Bacillus subtilis spores exposed to environmental solar UV radiation. AB - Spores of Bacillus subtilis contain a number of small, acid-soluble spore proteins (SASP) which comprise up to 20% of total spore core protein. The multiple alpha/beta-type SASP have been shown to confer resistance to UV radiation, heat, peroxides, and other sporicidal treatments. In this study, SASP defective mutants of B. subtilis and spores deficient in dacB, a mutation leading to an increased core water content, were used to study the relative contributions of SASP and increased core water content to spore resistance to germicidal 254-nm and simulated environmental UV exposure (280 to 400 nm, 290 to 400 nm, and 320 to 400 nm). Spores of strains carrying mutations in sspA, sspB, and both sspA and sspB (lacking the major SASP-alpha and/or SASP-beta) were significantly more sensitive to 254-nm and all polychromatic UV exposures, whereas the UV resistance of spores of the sspE strain (lacking SASP-gamma) was essentially identical to that of the wild type. Spores of the dacB-defective strain were as resistant to 254-nm UV-C radiation as wild-type spores. However, spores of the dacB strain were significantly more sensitive than wild-type spores to environmental UV treatments of >280 nm. Air-dried spores of the dacB mutant strain had a significantly higher water content than air-dried wild-type spores. Our results indicate that alpha/beta-type SASP and decreased spore core water content play an essential role in spore resistance to environmentally relevant UV wavelengths whereas SASP-gamma does not. PMID- 19542329 TI - Alleviation of proteolytic sensitivity to enhance recombinant lipase production in Escherichia coli. AB - Two amino acids, Leu149 and Val223, were identified as proteolytically sensitive when Pseudozyma antarctica lipase (PalB) was heterologously expressed in Escherichia coli. The functional expression was enhanced using the double mutant for cultivation. However, the recombinant protein production was still limited by PalB misfolding, which was resolved by DsbA coexpression. PMID- 19542330 TI - Significant enhanced expression and solubility of human proteins in Escherichia coli by fusion with protein S from Myxococcus xanthus. AB - Protein S is a major spore coat protein of Myxococcus xanthus, consisting of two homologous domains, the N-terminal domain (NTD) and the C-terminal domain, both of which contain a Ca(2+)-binding site. Protein S tightly binds to myxospores in a Ca(2+)-dependent manner. Here, we constructed a novel expression vector, pCold PST, encoding two tandem repeat NTDs (PrS2). By using this vector, a number of human proteins that were expressed at low levels or in insoluble forms by a pET vector were expressed not only at high levels but also in soluble forms. We also demonstrated that an Escherichia coli protein tagged with PrS2 fully retained its function, indicating that it is folded independently from the tag. This technology not only allows simple, one-step protein purification using myxospores, but can also be used for the identification of proteins interacting with a protein of interest and will prove immensely useful for structural studies of proteins which are difficult to produce or are insoluble. PMID- 19542331 TI - Role of P450 monooxygenases in the degradation of the endocrine-disrupting chemical nonylphenol by the white rot fungus Phanerochaete chrysosporium. AB - The white rot fungus Phanerochaete chrysosporium extensively degraded the endocrine disruptor chemical nonylphenol (NP; 100% of 100 ppm) in both nutrient limited cultures and nutrient-sufficient cultures. The P450 enzyme inhibitor piperonyl butoxide caused significant inhibition (approximately 75%) of the degradation activity in nutrient-rich malt extract (ME) cultures but no inhibition in defined low-nitrogen (LN) cultures, indicating an essential role of P450 monooxygenase(s) in NP degradation under nutrient-rich conditions. A genome wide analysis using our custom-designed P450 microarray revealed significant induction of multiple P450 monooxygenase genes by NP: 18 genes were induced (2- to 195-fold) under nutrient-rich conditions, 17 genes were induced (2- to 6-fold) in LN cultures, and 3 were induced under both nutrient-rich and LN conditions. The P450 genes Pff 311b (corresponding to protein identification number [ID] 5852) and Pff 4a (protein ID 5001) showed extraordinarily high levels of induction (195- and 167-fold, respectively) in ME cultures. The P450 oxidoreductase (POR), glutathione S-transferase (gst), and cellulose metabolism genes were also induced in ME cultures. In contrast, certain metabolic genes, such as five of the peroxidase genes, showed partial downregulation by NP. This study provides the first evidence for the involvement of P450 enzymes in NP degradation by a white rot fungus and the first genome-wide identification of specific P450 genes responsive to an environmentally significant toxicant. PMID- 19542333 TI - Plasmid-based system for high-level gene expression and antisense gene knockdown in Bartonella henselae. AB - Six broad-host-range plasmid vectors were developed to study gene expression in Bartonella henselae. The vectors were used to express a beta-galactosidase reporter gene in B. henselae and to generate antisense RNA for gene knockdown. When applied to ompR, a putative transcription response regulator of B. henselae, this antisense RNA gene knockdown strategy reduced bacterial invasion of human endothelial cells by over 60%. PMID- 19542334 TI - Prevalence of Escherichia coli O157:H7 in organically and naturally raised beef cattle. AB - We determined the prevalence of Escherichia coli O157:H7 in organically and naturally raised beef cattle at slaughter and compared antibiotic susceptibility profiles of the isolates to those of isolates from conventionally raised beef cattle. The prevalences of E. coli O157:H7 were 14.8 and 14.2% for organically and naturally raised cattle, respectively. No major difference in antibiotic susceptibility patterns among the isolates was observed. PMID- 19542332 TI - A new Borrelia species defined by multilocus sequence analysis of housekeeping genes. AB - Analysis of Lyme borreliosis (LB) spirochetes, using a novel multilocus sequence analysis scheme, revealed that OspA serotype 4 strains (a rodent-associated ecotype) of Borrelia garinii were sufficiently genetically distinct from bird associated B. garinii strains to deserve species status. We suggest that OspA serotype 4 strains be raised to species status and named Borrelia bavariensis sp. nov. The rooted phylogenetic trees provide novel insights into the evolutionary history of LB spirochetes. PMID- 19542335 TI - Role of growth temperature in freeze-thaw tolerance of Listeria spp. AB - The food-borne pathogen Listeria monocytogenes can grow in a wide range of temperatures, and several key virulence determinants of the organism are expressed at 37 degrees C but are strongly repressed below 30 degrees C. However, the impact of growth temperature on the ability of the bacteria to tolerate environmental stresses remains poorly understood. In other microorganisms, cold acclimation resulted in enhanced tolerance against freezing and thawing (cryotolerance). In this study, we investigated the impact of growth temperature (4, 25, and 37 degrees C) on the cryotolerance of 14 strains of L. monocytogenes from outbreaks and from food processing plant environments and four strains of nonpathogenic Listeria spp. (L. welshimeri and L. innocua). After growth at different temperatures, cells were frozen at -20 degrees C, and repeated freeze thaw cycles were applied every 24 h. Pronounced cryotolerance was exhibited by cells grown at 37 degrees C, with a <1-log decrease after 18 cycles of freezing and thawing. In contrast, freeze-thaw tolerance was significantly reduced (P < 0.05) when bacteria were grown at either 4 or 25 degrees C, with log decreases after 18 freeze-thaw cycles ranging from 2 to >4, depending on the strain. These findings suggest that growth at 37 degrees C, a temperature required for expression of virulence determinants of L. monocytogenes, is also required for protection against freeze-thaw stress. The negative impact of growth at low temperature on freeze-thaw stress was unexpected and has not been reported before with this or other psychrotrophic microorganisms. PMID- 19542336 TI - Chickens and cattle as sources of sporadic domestically acquired Campylobacter jejuni infections in Finland. AB - A substantial sampling among domestic human campylobacter cases, chicken process lots, and cattle at slaughter was performed during the seasonal peak of human infections. Campylobacter jejuni isolates (n = 419) were subtyped using pulsed field gel electrophoresis with SmaI, and isolates representing overlapping types (n = 212) were further subtyped using KpnI for restriction. The SmaI/KpnI profiles of 55.4% (97/175) of the human isolates were indistinguishable from those of the chicken or cattle isolates. The overlapping SmaI/KpnI subtypes accounted for 69.8% (30/43) and 15.9% (32/201) of the chicken and cattle isolates, respectively. The occurrence of identical SmaI/KpnI subtypes with human C. jejuni isolates was significantly associated with animal host species (P < 0.001). A temporal association of isolates from chickens and patients was possible in 31.4% (55/175) of the human infections. Besides chickens as sources of C. jejuni in the sporadic infections, the role of cattle appears notable. New approaches to restrict the occurrence of campylobacters in other farm animals may be needed in addition to hygienic measures in chicken production. However, only about half of the human infections were attributable to these sources. PMID- 19542337 TI - Energy taxis drives Campylobacter jejuni toward the most favorable conditions for growth. AB - Campylobacter jejuni is a serious food-borne bacterial pathogen in the developed world. Poultry is a major reservoir, and C. jejuni appears highly adapted to the gastrointestinal tract of birds. Several factors are important for chicken colonization and virulence, including a taxis mechanism for environmental navigation. To explore the mechanism of chemotaxis in C. jejuni, we constructed mutants with deletions of five putative mcp (methyl-accepting chemotaxis protein) genes (tlp1, tlp2, tlp3, docB, and docC). Surprisingly, the deletions did not affect the chemotactic behavior of the mutants compared to that of the parental strain. However, the tlp1, tlp3, docB, and docC mutant strains displayed a 10 fold decrease in the ability to invade human epithelial and chicken embryo cells, hence demonstrating that the corresponding proteins affect the host interaction. l-Asparagine, formate, d-lactate, and chicken mucus were identified as new attractants of C. jejuni, and we observed that chemical substances promoting tactic attraction are all known to support the growth of this organism. The attractants could be categorized as carbon sources and electron donors and acceptors, and we furthermore observed a correlation between an attractant's potency and its efficiency as an energy source. The tactic attraction was inhibited by the respiratory inhibitors HQNO (2-n-heptyl-4-hydroxyquinoline N oxide) and sodium azide, which significantly reduce energy production by oxidative phosphorylation. These findings strongly indicate that energy taxis is the primary force in environmental navigation by C. jejuni and that this mechanism drives the organism toward the optimal chemical conditions for energy generation and colonization. PMID- 19542338 TI - Evolution of Lactococcus lactis phages within a cheese factory. AB - We have sequenced the double-stranded DNA genomes of six lactococcal phages (SL4, CB13, CB14, CB19, CB20, and GR7) from the 936 group that were isolated over a 9 year period from whey samples obtained from a Canadian cheese factory. These six phages infected the same two industrial Lactococcus lactis strains out of 30 tested. The CB14 and GR7 genomes were found to be 100% identical even though they were isolated 14 months apart, indicating that a phage can survive in a cheese plant for more than a year. The other four genomes were related but notably different. The length of the genomes varied from 28,144 to 32,182 bp, and they coded for 51 to 55 open reading frames. All five genomes possessed a 3' overhang cos site that was 11 nucleotides long. Several structural proteins were also identified by nano-high-performance liquid chromatography-tandem mass spectrometry, confirming bioinformatic analyses. Comparative analyses suggested that the most recently isolated phages (CB19 and CB20) were derived, in part, from older phage isolates (CB13 and CB14/GR7). The organization of the five distinct genomes was similar to the previously sequenced lactococcal phage genomes of the 936 group, and from these sequences, a core genome was determined for lactococcal phages of the 936 group. PMID- 19542339 TI - Inactivation of PadR, the repressor of the phenolic acid stress response, by molecular interaction with Usp1, a universal stress protein from Lactobacillus plantarum, in Escherichia coli. AB - The phenolic acid decarboxylase gene padA is involved in the phenolic acid stress response (PASR) in gram-positive bacteria. In Lactobacillus plantarum, the padR gene encodes the negative transcriptional regulator of padA and is cotranscribed with a downstream gene, usp1, which encodes a putative universal stress protein (USP), Usp1, of unknown function. The usp1 gene is overexpressed during the PASR. However, the role and the mechanism of action of the USPs are unknown in gram positive bacteria. Therefore, to gain insights into the role of USPs in the PASR; (i) a usp1 deletion mutant was constructed; (ii) the two genes padR and usp1 were coexpressed with padA under its own promoter as a reporter gene in Escherichia coli; and (iii) molecular in vitro interactions between the PadR, Usp1, and the padA promoter were studied. Although the usp1 mutant strain retained phenolic acid-dependent PAD activity, it displayed a greater sensitivity to strong acidic conditions compared to that of the wild-type strain. PadR cannot be inactivated directly by phenolic acid in E. coli recombinant cultures but is inactivated by Usp1 when the two proteins are coexpressed in E. coli. The PadR inactivation observed in recombinant E. coli cells was supported by electrophoretic mobility shift assays. Although Usp1 seems not to be absolutely required for the PASR, its capacity to inactivate PadR indicates that it could serve as an important mediator in acid stress response mechanisms through its capacity to interact with transcriptional regulators. PMID- 19542340 TI - Controlled gene expression in the plant pathogen Cryphonectria parasitica by use of a copper-responsive element. AB - We have developed a tool for controlled expression of heterologous or ectopic genes in the chestnut pathogen Cryphonectria parasitica using the promoter region from a putative copper-regulated transporter gene. In addition, we have found that expression control via this system is not affected by the virulence attenuating hypovirus CHV1-EP713. PMID- 19542342 TI - Kinetic characterization of OmcA and MtrC, terminal reductases involved in respiratory electron transfer for dissimilatory iron reduction in Shewanella oneidensis MR-1. AB - We have used scaling kinetics and the concept of kinetic competence to elucidate the role of hemeproteins OmcA and MtrC in iron reduction by Shewanella oneidensis MR-1. Second-order rate constants for OmcA and MtrC were determined by single turnover experiments. For soluble iron species, a stopped-flow apparatus was used, and for the less reactive iron oxide goethite, a conventional spectrophotometer was used to measure rates. Steady-state experiments were performed to obtain molecular rate constants by quantifying the OmcA and MtrC contents of membrane fractions and whole cells by Western blot analysis. For reduction of soluble iron, rates determined from transient-state experiments were able to account for rates obtained from steady-state experiments. However, this was not true with goethite; rate constants determined from transient-state experiments were 100 to 1,000 times slower than those calculated from steady state experiments with membrane fractions and whole cells. In contrast, addition of flavins to the goethite experiments resulted in rates that were consistent with both transient- and steady-state experiments. Kinetic simulations of steady state results with kinetic constants obtained from transient-state experiments supported flavin involvement. Therefore, we show for the first time that OmcA and MtrC are kinetically competent to account for catalysis of soluble iron reduction in whole Shewanella cells but are not responsible for electron transfer via direct contact alone with insoluble iron-containing minerals. This work supports the hypothesis that electron shuttles are important participants in the reduction of solid Fe phases by this organism. PMID- 19542341 TI - Bacterial and archaeal phylogenetic diversity of a cold sulfur-rich spring on the shoreline of Lake Erie, Michigan. AB - Studies of sulfidic springs have provided new insights into microbial metabolism, groundwater biogeochemistry, and geologic processes. We investigated Great Sulphur Spring on the western shore of Lake Erie and evaluated the phylogenetic affiliations of 189 bacterial and 77 archaeal 16S rRNA gene sequences from three habitats: the spring origin (11-m depth), bacterial-algal mats on the spring pond surface, and whitish filamentous materials from the spring drain. Water from the spring origin water was cold, pH 6.3, and anoxic (H(2), 5.4 nM; CH(4), 2.70 microM) with concentrations of S(2-) (0.03 mM), SO(4)(2-) (14.8 mM), Ca(2+) (15.7 mM), and HCO(3)(-) (4.1 mM) similar to those in groundwater from the local aquifer. No archaeal and few bacterial sequences were >95% similar to sequences of cultivated organisms. Bacterial sequences were largely affiliated with sulfur metabolizing or chemolithotrophic taxa in Beta-, Gamma-, Delta-, and Epsilonproteobacteria. Epsilonproteobacteria sequences similar to those obtained from other sulfidic environments and a new clade of Cyanobacteria sequences were particularly abundant (16% and 40%, respectively) in the spring origin clone library. Crenarchaeota sequences associated with archaeal-bacterial consortia in whitish filaments at a German sulfidic spring were detected only in a similar habitat at Great Sulphur Spring. This study expands the geographic distribution of many uncultured Archaea and Bacteria sequences to the Laurentian Great Lakes, indicates possible roles for epsilonproteobacteria in local aquifer chemistry and karst formation, documents new oscillatorioid Cyanobacteria lineages, and shows that uncultured, cold-adapted Crenarchaeota sequences may comprise a significant part of the microbial community of some sulfidic environments. PMID- 19542343 TI - Novel denitrifying bacterium Ochrobactrum anthropi YD50.2 tolerates high levels of reactive nitrogen oxides. AB - Most studies of bacterial denitrification have used nitrate (NO3-) as the first electron acceptor, whereas relatively less is understood about nitrite (NO2-) denitrification. We isolated novel bacteria that proliferated in the presence of high levels of NO2- (72 mM). Strain YD50.2, among several isolates, was taxonomically positioned within the alpha subclass of Proteobacteria and identified as Ochrobactrum anthropi YD50.2. This strain denitrified NO2-, as well as NO3-. The gene clusters for denitrification (nar, nir, nor, and nos) were cloned from O. anthropi YD50.2, in which the nir and nor operons were linked. We confirmed that nirK in the nir-nor operon produced a functional NO2- reductase containing copper that was involved in bacterial NO2- reduction. The strain denitrified up to 40 mM NO2- to dinitrogen under anaerobic conditions in which other denitrifiers or NO3- reducers such as Pseudomonas aeruginosa and Ralstonia eutropha and nitrate-respiring Escherichia coli neither proliferated nor reduced NO2-. Under nondenitrifying aerobic conditions, O. anthropi YD50.2 and its type strain ATCC 49188(T) proliferated even in the presence of higher levels of NO2- (100 mM), and both were considerably more resistant to acidic NO2- than were the other strains noted above. These results indicated that O. anthropi YD50.2 is a novel denitrifier that has evolved reactive nitrogen oxide tolerance mechanisms. PMID- 19542344 TI - Bacillus thuringiensis bel protein enhances the toxicity of Cry1Ac protein to Helicoverpa armigera larvae by degrading insect intestinal mucin. AB - Bacillus thuringiensis has been used as a bioinsecticide to control agricultural insects. Bacillus cereus group genomes were found to have a Bacillus enhancin like (bel) gene, encoding a peptide with 20 to 30% identity to viral enhancin protein, which can enhance viral infection by degradation of the peritrophic matrix (PM) of the insect midgut. In this study, the bel gene was found to have an activity similar to that of the viral enhancin gene. A bel knockout mutant was constructed by using a plasmid-free B. thuringiensis derivative, BMB171. The 50% lethal concentrations of this mutant plus the cry1Ac insecticidal protein gene were about 5.8-fold higher than those of the BMB171 strain. When purified Bel was mixed with the Cry1Ac protein and fed to Helicoverpa armigera larvae, 3 mug/ml Cry1Ac alone induced 34.2% mortality. Meanwhile, the mortality rate rose to 74.4% when the same amount of Cry1Ac was mixed with 0.8 mug/ml of Bel. Microscopic observation showed a significant disruption detected on the midgut PM of H. armigera larvae after they were fed Bel. In vitro degradation assays showed that Bel digested the intestinal mucin (IIM) of Trichoplusia ni and H. armigera larvae to various degrading products, similar to findings for viral enhancin. These results imply Bel toxicity enhancement depends on the destruction of midgut PM and IIM, similar to the case with viral enhancin. This discovery showed that Bel has the potential to enhance insecticidal activity of B. thuringiensis-based biopesticides and transgenic crops. PMID- 19542345 TI - Comparative genome-wide transcriptional profiling of Azorhizobium caulinodans ORS571 grown under free-living and symbiotic conditions. AB - The whole-genome sequence of the endosymbiotic bacterium Azorhizobium caulinodans ORS571, which forms nitrogen-fixing nodules on the stems and roots of Sesbania rostrata, was recently determined. The sizes of the genome and symbiosis island are 5.4 Mb and 86.7 kb, respectively, and these sizes are the smallest among the sequenced rhizobia. In the present study, a whole-genome microarray of A. caulinodans was constructed, and transcriptomic analyses were performed on free living cells grown in rich and minimal media and in bacteroids isolated from stem nodules. Transcriptional profiling showed that the genes involved in sulfur uptake and metabolism, acetone metabolism, and the biosynthesis of exopolysaccharide were highly expressed in bacteroids compared to the expression levels in free-living cells. Some mutants having Tn5 transposons within these genes with increased expression were obtained as nodule-deficient mutants in our previous study. A transcriptomic analysis was also performed on free-living cells grown in minimal medium supplemented with a flavonoid, naringenin, which is one of the most efficient inducers of A. caulinodans nod genes. Only 18 genes exhibited increased expression by the addition of naringenin, suggesting that the regulatory mechanism responding to the flavonoid could be simple in A. caulinodans. The combination of our genome-wide transcriptional profiling and our previous genome-wide mutagenesis study has revealed new aspects of nodule formation and maintenance. PMID- 19542346 TI - Aerobic biodegradation of N-nitrosodimethylamine by the propanotroph Rhodococcus ruber ENV425. AB - The propanotroph Rhodococcus ruber ENV425 was observed to rapidly biodegrade N nitrosodimethylamine (NDMA) after growth on propane, tryptic soy broth, or glucose. The key degradation intermediates were methylamine, nitric oxide, nitrite, nitrate, and formate. Small quantities of formaldehyde and dimethylamine were also detected. A denitrosation reaction, initiated by hydrogen atom abstraction from one of the two methyl groups, is hypothesized to result in the formation of n-methylformaldimine and nitric oxide, the former of which decomposes in water to methylamine and formaldehyde and the latter of which is then oxidized further to nitrite and then nitrate. Although the strain mineralized more than 60% of the carbon in [(14)C]NDMA to (14)CO(2), growth of strain ENV425 on NDMA as a sole carbon and energy source could not be confirmed. The bacterium was capable of utilizing NDMA, as well as the degradation intermediates methylamine and nitrate, as sources of nitrogen during growth on propane. In addition, ENV425 reduced environmentally relevant microgram/liter concentrations of NDMA to <2 ng/liter in batch cultures, suggesting that the bacterium may have applications for groundwater remediation. PMID- 19542347 TI - Comparative metagenomic analysis of a microbial community residing at a depth of 4,000 meters at station ALOHA in the North Pacific subtropical gyre. AB - The deep sea (water depth of >2,000 m) represents the largest biome on Earth. Yet relatively little is known about its microbial community's structure, function, and adaptation to the cold and deep biosphere. To provide further genomic insights into deep-sea planktonic microbes, we sequenced a total of approximately 200 Mbp of a random whole-genome shotgun (WGS) library from a microbial community residing at a depth of 4,000 m at Station ALOHA in the Pacific Ocean and compared it to other available WGS sequence data from surface and deep waters. Our analyses indicated that the deep-sea lifestyle is likely facilitated by a collection of very subtle adaptations, as opposed to dramatic alterations of gene content or structure. These adaptations appear to include higher metabolic versatility and genomic plasticity to cope with the sparse and sporadic energy resources available, a preference for hydrophobic and smaller-volume amino acids in protein sequences, unique proteins not found in surface-dwelling species, and adaptations at the gene expression level. The deep-sea community is also characterized by a larger average genome size and a higher content of "selfish" genetic elements, such as transposases and prophages, whose propagation is apparently favored by more relaxed purifying (negative) selection in deeper waters. PMID- 19542348 TI - Identification of three alcohol dehydrogenase genes involved in the stereospecific catabolism of arylglycerol-beta-aryl ether by Sphingobium sp. strain SYK-6. AB - Degradation of arylglycerol-beta-aryl ether is the most important process in bacterial lignin catabolism. Sphingobium sp. strain SYK-6 degrades guaiacylglycerol-beta-guaiacyl ether (GGE) to alpha-(2-methoxyphenoxy)-beta hydroxypropiovanillone (MPHPV), and then the ether linkage of MPHPV is cleaved to generate alpha-glutathionyl-beta-hydroxypropiovanillone (GS-HPV) and guaiacol. We have characterized three enantioselective glutathione S-transferase genes, including two genes that are involved in the ether cleavage of two enantiomers of MPHPV and one gene that is involved in the elimination of glutathione from a GS HPV enantiomer. However, the first step in the degradation of four different GGE stereoisomers has not been characterized. In this study, three alcohol dehydrogenase genes, ligL, ligN, and ligO, which conferred GGE transformation activity in Escherichia coli, were isolated from SYK-6 and characterized, in addition to the previously cloned ligD gene. The levels of amino acid sequence identity of the four GGE dehydrogenases, which belong to the short-chain dehydrogenase/reductase family, ranged from 32% to 39%. Each gene was expressed in E. coli, and the stereospecificities of the gene products with the four GGE stereoisomers were determined by using chiral high-performance liquid chromatography with recently synthesized authentic enantiopure GGE stereoisomers. LigD and LigO converted (alphaR,betaS)-GGE and (alphaR,betaR)-GGE into (betaS) MPHPV and (betaR)-MPHPV, respectively, while LigL and LigN transformed (alphaS,betaR)-GGE and (alphaS,betaS)-GGE to (betaR)-MPHPV and (betaS)-MPHPV, respectively. Disruption of the genes indicated that ligD is essential for the degradation of (alphaR,betaS)-GGE and (alphaR,betaR)-GGE and that both ligL and ligN contribute to the degradation of the two other GGE stereoisomers. PMID- 19542350 TI - Why elementary teachers might be inadequately prepared to teach reading. AB - Several national reports have suggested the usefulness of systematic, explicit, synthetic phonics instruction based on English word structure along with wide reading of quality literature for supporting development in early reading instruction. Other studies have indicated, however, that many in-service teachers are not knowledgeable in the basic concepts of the English language. They may be well versed in children's literature but not know how to address the basic building blocks of language and reading. The authors hypothesized that one of the reasons for this situation is that many instructors responsible for training future elementary teachers are not familiar with the concepts of the linguistic features of English language. This hypothesis was tested by administering a survey of language concepts to 78 instructors. The results showed that even though teacher educators were familiar with syllabic knowledge, they performed poorly on concepts relating to morphemes and phonemes. In a second study, 40 instructors were interviewed about best practices in teaching components and subskills of reading. Eighty percent of instructors defined phonological awareness as letter-sound correspondence. They also did not mention synthetic phonics as a desirable method to use for beginning reading instruction, particularly for students at risk for reading difficulties. In conclusion, providing professional development experiences related to language concepts to instructors could provide them the necessary knowledge of language concepts related to early literacy instruction, which they could then integrate into their preservice reading courses. PMID- 19542349 TI - Evolution and diversity of facultative symbionts from the aphid subfamily Lachninae. AB - Many aphids harbor a variety of endosymbiotic bacteria. The functions of these symbionts can range from an obligate nutritional role to a facultative role in protecting their hosts against environmental stresses. One such symbiont is "Candidatus Serratia symbiotica," which is involved in defense against heat and potentially also in aphid nutrition. Lachnid aphids have been the focus of several recent studies investigating the transition of this symbiont from a facultative symbiont to an obligate symbiont. In a phylogenetic analysis of Serratia symbionts from 51 lachnid hosts, we found that diversity in symbiont morphology, distribution, and function is due to multiple independent origins of symbiosis from ancestors belonging to Serratia and possibly also to evolution within distinct symbiont clades. Our results do not support cocladogenesis of "Ca. Serratia symbiotica" with Cinara subgenus Cinara species and weigh against an obligate nutritional role. Finally, we show that species belonging to the subfamily Lachninae have a high incidence of facultative symbiont infection. PMID- 19542353 TI - Cell rearrangements, cell divisions and cell death in a migrating epithelial sheet in the abdomen of Drosophila. AB - During morphogenesis, cell movements, cell divisions and cell death work together to form complex patterns and to shape organs. These events are the outcome of decisions made by many individual cells, but how these decisions are controlled and coordinated is elusive. The adult abdominal epidermis of Drosophila is formed during metamorphosis by divisions and extensive cell migrations of the diploid histoblasts, which replace the polyploid larval cells. Using in vivo 4D microscopy, we have studied the behaviour of the histoblasts and analysed in detail how they reach their final position and to what extent they rearrange during their spreading. Tracking individual cells, we show that the cells migrate in two phases that differ in speed, direction and amount of cellular rearrangement. Cells of the anterior (A) and posterior (P) compartments differ in their behaviour. Cells near the A/P border are more likely to change their neighbours during migration. The mitoses do not show any preferential orientation. After mitosis, the sisters become preferentially aligned with the direction of movement. Thus, in the abdomen, it is the extensive cell migrations that appear to contribute most to morphogenesis. This contrasts with other developing epithelia, such as the wing imaginal disc and the embryonic germband in Drosophila, where oriented mitoses and local cell rearrangements appear to direct morphogenesis. Furthermore, our results suggest that an active force created by the histoblasts contributes to the formation of the adult epidermis. Finally, we show that histoblasts occasionally undergo apoptosis. PMID- 19542351 TI - The transcriptional foundation of pluripotency. AB - A fundamental goal in biology is to understand the molecular basis of cell identity. Pluripotent embryonic stem (ES) cell identity is governed by a set of transcription factors centred on the triumvirate of Oct4, Sox2 and Nanog. These proteins often bind to closely localised genomic sites. Recent studies have identified additional transcriptional modulators that bind to chromatin near sites occupied by Oct4, Sox2 and Nanog. This suggests that the combinatorial control of gene transcription might be fundamental to the ES cell state. Here we discuss how these observations advance our understanding of the transcription factor network that controls pluripotent identity and highlight unresolved issues that arise from these studies. PMID- 19542352 TI - Cux2 functions downstream of Notch signaling to regulate dorsal interneuron formation in the spinal cord. AB - Obtaining the diversity of interneuron subtypes in their appropriate numbers requires the orchestrated integration of progenitor proliferation with the regulation of differentiation. Here we demonstrate through loss-of-function studies in mice that the Cut homeodomain transcription factor Cux2 (Cutl2) plays an important role in regulating the formation of dorsal spinal cord interneurons. Furthermore, we show that Notch regulates Cux2 expression. Although Notch signaling can be inhibitory to the expression of proneural genes, it is also required for interneuron formation during spinal cord development. Our findings suggest that Cux2 might mediate some of the effects of Notch signaling on interneuron formation. Together with the requirement for Cux2 in cell cycle progression, our work highlights the mechanistic complexity in balancing neural progenitor maintenance and differentiation during spinal cord neurogenesis. PMID- 19542355 TI - LATERAL ORGAN FUSION1 and LATERAL ORGAN FUSION2 function in lateral organ separation and axillary meristem formation in Arabidopsis. AB - Plant organs are generated from meristems throughout development. Patterning and elaboration of organ primordia occur as a result of organized cell division and expansion, processes that are likely to be controlled, in part, by meristem derived signals. Communication between the meristem and lateral organs is crucial for meristem maintenance and organ patterning, and organ boundaries are thought to be important for mediating this communication. Arabidopsis thaliana LATERAL ORGAN FUSION1 (LOF1) encodes a MYB-domain transcription factor that is expressed in organ boundaries. lof1 mutants display defects in organ separation as a result of abnormal cell division and expansion during early boundary formation. lof1 mutants also fail to form accessory shoot meristems. Mutations in the closely related LATERAL ORGAN FUSION2 (LOF2) gene enhance the lof1 phenotype, such that lof1 lof2 double mutants display additional fusion defects. Genetic interactions with the CUP-SHAPED COTYLEDON genes CUC2 and CUC3 revealed a role for LOF1 in both organ separation and axillary meristem formation. Expression of the meristem determinant STM was reduced in lof1 mutant paraclade junctions and lof1 enhanced the weak stm-10 mutant, such that double mutants had severe defects in meristem maintenance and organ separation. Our data implicate LOF1 and LOF2 in boundary specification, meristem initiation and maintenance, and organ patterning. PMID- 19542354 TI - The RNA-binding protein Mex3b has a fine-tuning system for mRNA regulation in early Xenopus development. AB - Post-transcriptional control by RNA-binding proteins is a precise way to assure appropriate levels of gene expression. Here, we identify a novel mRNA regulatory system involving Mex3b (RKHD3) and demonstrate its role in FGF signaling. mex3b mRNA has a 3' long conserved UTR, named 3'LCU, which contains multiple elements for both mRNA destabilization and translational enhancement. Notably, Mex3b promotes destabilization of its own mRNA by binding to the 3'LCU, thereby forming a negative autoregulatory loop. The combination of positive regulation and negative autoregulation constitutes a fine-tuning system for post-transcriptional control. In early embryogenesis, Mex3b is involved in anteroposterior patterning of the neural plate. Consistent with this, Mex3b can attenuate FGF signaling and destabilize mRNAs for the FGF signaling components Syndecan 2 and Ets1b through their 3' UTRs. These data suggest that the 3'LCU-mediated fine-tuning system determines the appropriate level of mex3b expression, which in turn contributes to neural patterning through regulating FGF signaling. PMID- 19542356 TI - Regulation of stem cell maintenance by the Polycomb protein FIE has been conserved during land plant evolution. AB - The Polycomb group (PcG) complex is involved in the epigenetic control of gene expression profiles. In flowering plants, PcG proteins regulate vegetative and reproductive programs. Epigenetically inherited states established in the gametophyte generation are maintained after fertilization in the sporophyte generation, having a profound influence on seed development. The gametophyte size and phase dominance were dramatically reduced during angiosperm evolution, and have specialized in flowering plants to support the reproductive process. The moss Physcomitrella patens is an ideal organism in which to study epigenetic processes during the gametophyte stage, as it possesses a dominant photosynthetic gametophytic haploid phase and efficient homologous recombination, allowing targeted gene replacement. We show that P. patens PcG protein FIE (PpFIE) accumulates in haploid meristematic cells and in cells that undergo fate transition during dedifferentiation programs in the gametophyte. In the absence of PpFIE, meristems overproliferate and are unable to develop leafy gametophytes or reach the reproductive phase. This aberrant phenotype might result from failure of the PcG complex to repress proliferation and differentiation of three faced apical stem cells, which are designated to become lateral shoots. The PpFIE phenotype can be partially rescued by FIE of Arabidopsis thaliana, a flowering plant that diverged >450 million years ago from bryophytes. PpFIE can partially complement the A. thaliana fie mutant, illustrating functional conservation of the protein during evolution in regulating the differentiation of meristematic cells in gametophyte development, both in bryophytes and angiosperms. This mechanism was harnessed at the onset of the evolution of alternating generations, facilitating the establishment of sporophytic developmental programs. PMID- 19542357 TI - C. elegans Rab GTPase activating protein TBC-2 promotes cell corpse degradation by regulating the small GTPase RAB-5. AB - During apoptosis, dying cells are quickly internalized by neighboring cells or phagocytes, and are enclosed in phagosomes that undergo a maturation process to generate the phagoslysosome, in which cell corpses are eventually degraded. It is not well understood how apoptotic cell degradation is regulated. Here we report the identification and characterization of the C. elegans tbc-2 gene, which is required for the efficient degradation of cell corpses. tbc-2 encodes a Rab GTPase activating protein (GAP) and its loss of function affects several events of phagosome maturation, including RAB-5 release, phosphatidylinositol 3 phosphate dynamics, phagosomal acidification, RAB-7 recruitment and lysosome incorporation, which leads to many persistent cell corpses at various developmental stages. Intriguingly, the persistent cell corpse phenotype of tbc-2 mutants can be suppressed by reducing gene expression of rab-5, and overexpression of a GTP-locked RAB-5 caused similar defects in phagosome maturation and cell corpse degradation. We propose that TBC-2 functions as a GAP to cycle RAB-5 from an active GTP-bound to an inactive GDP-bound state, which is required for maintaining RAB-5 dynamics on phagosomes and serves as a switch for the progression of phagosome maturation. PMID- 19542358 TI - FGF signaling is strictly required to maintain early telencephalic precursor cell survival. AB - The FGF family of extracellular signaling factors has been proposed to play multiple roles in patterning the telencephalon, the precursor to the cerebrum. In this study, unlike previous ones, we effectively abolish FGF signaling in the anterior neural plate via deletion of three FGF receptor (FGFR) genes. Triple FGFR mutant mice exhibit a complete loss of the telencephalon, except the dorsal midline. Disruption of FGF signaling prior to and coincident with telencephalic induction reveals that FGFs promote telencephalic character and are strictly required to keep telencephalic cells alive. Moreover, progressively more severe truncations of the telencephalon are observed in FGFR single, double and triple mutants. Together with previous gain-of-function studies showing induction of Foxg1 expression and mirror-image duplications of the cortex by exogenous FGF8, our loss-of-function results suggest that, rather than independently patterning different areas, FGF ligands and receptors act in concert to mediate organizer activity for the whole telencephalon. PMID- 19542359 TI - Promotion of proliferation in the developing cerebral cortex by EphA4 forward signaling. AB - Eph receptors are widely expressed during cerebral cortical development, yet a role for Eph signaling in the generation of cells during corticogenesis has not been shown. Cortical progenitor cells selectively express one receptor, EphA4, and reducing EphA4 signaling in cultured progenitors suppressed proliferation, decreasing cell number. In vivo, EphA4(-/-) cortex had a reduced area, fewer cells and less cell division compared with control cortex. To understand the effects of EphA4 signaling in corticogenesis, EphA4-mediated signaling was selectively depressed or elevated in cortical progenitors in vivo. Compared with control cells, cells with reduced EphA4 signaling were rare and mitotically inactive. Conversely, overexpression of EphA4 maintained cells in their progenitor states at the expense of subsequent maturation, enlarging the progenitor pool. These results support a role for EphA4 in the autonomous promotion of cell proliferation during corticogenesis. Although most ephrins were undetectable in cortical progenitors, ephrin B1 was highly expressed. Our analyses demonstrate that EphA4 and ephrin B1 bind to each other, thereby initiating signaling. Furthermore, overexpression of ephrin B1 stimulated cell division of neighboring cells, supporting the hypothesis that ephrin B1-initiated forward signaling of EphA4 promotes cortical cell division. PMID- 19542361 TI - Evaluating the claim of enhanced persistence: the case of osteoporosis and implications for payers. AB - Cost-effectiveness analysis (CEA) has been widely used in evaluating treatments for osteoporosis. To study the claim of enhanced persistence, this research determined the effects of persistence (the proportion of individuals who remain on treatment) and efficacy on incremental cost-effectiveness ratios (ICERs) for bisphosphonate treatment relative to no bisphosphonate treatment in the United States. For 2 age groups, 55 to 59 and 75 to 79, the authors relied on published fracture rates and applied them to 1000 postmenopausal osteoporotic patients with bone mineral density (BMD) T score < or = -2.5 during 3 years of treatment. After developing an algebraic ICER, with effectiveness measured by either quality adjusted life years (QALYs) gained or number of fractures averted, they determined the effects of persistence and efficacy and then calibrated the model to variable estimates from the literature. For the younger (older) cohort, the cost per fracture averted was $66,606 ($18,256), consistent with a validated Markov simulation model. The effect of a 1 percentage point change in vertebral efficacy was 24 (5) times the effect of a 1 percentage point change in persistence for the younger cohort when QALYs (fractures) were involved. Nonvertebral efficacy had approximately 27 (9) times the effect of persistence. For the older cohort, the ratios were 15 (4.5) and 33 (10) for vertebral and nonvertebral fractures, respectively. In evaluating the claim of enhanced persistence, formulary decision makers need to exercise caution to ensure that efficacy is not compromised. Two drugs would have to be virtually identical in efficacy for better persistence to improve cost-effectiveness. PMID- 19542360 TI - Insm1 (IA-1) is an essential component of the regulatory network that specifies monoaminergic neuronal phenotypes in the vertebrate hindbrain. AB - Monoaminergic neurons include the physiologically important central serotonergic and noradrenergic subtypes. Here, we identify the zinc-finger transcription factor, Insm1, as a crucial mediator of the differentiation of both subtypes, and in particular the acquisition of their neurotransmitter phenotype. Insm1 is expressed in hindbrain progenitors of monoaminergic neurons as they exit the cell cycle, in a pattern that partially overlaps with the expression of the proneural factor Ascl1. Consistent with this, a conserved cis-regulatory sequence associated with Insm1 is bound by Ascl1 in the hindbrain, and Ascl1 is essential for the expression of Insm1 in the ventral hindbrain. In Insm1-null mutant mice, the expression of the serotonergic fate determinants Pet1, Lmx1b and Gata2 is markedly downregulated. Nevertheless, serotonergic precursors begin to differentiate in Insm1 mutants, but fail to produce serotonin because of a failure to activate expression of tryptophan hydroxylase 2 (Tph2), the key enzyme of serotonin biosynthesis. We find that both Insm1 and Ascl1 coordinately specify Tph2 expression. In brainstem noradrenergic centres of Insm1 mutants, expression of tyrosine hydroxylase is delayed in the locus coeruleus and is markedly deficient in the medullary noradrenergic nuclei. However, Insm1 is dispensable for the expression of a second key noradrenergic biosynthetic enzyme, dopamine beta-hydroxylase, which is instead regulated by Ascl1. Thus, Insm1 regulates the synthesis of distinct monoaminergic neurotransmitters by acting combinatorially with, or independently of, Ascl1 in specific monoaminergic populations. PMID- 19542362 TI - Successful treatment of bacterial infection hinders development of acquired immunity. AB - Antibiotics are routinely used to control bacterial infection, but the acquisition of acquired immunity following successful treatment has rarely been examined. We developed a model that allows visualization of acquired immunity during and following antibiotic treatment of typhoid. Pathogen-specific humoral and cellular immune responses were activated rapidly in antibiotic-treated mice, but were not sustained after successful antibiotic treatment and did not confer protection to secondary infection. In marked contrast, pathogen-specific Th1 and Ab responses matured over several weeks following immunization with a live vaccine strain. The deficiency in protective immunity following antibiotic treatment could be overcome by administering flagellin during antibiotic therapy. Thus, development of protective immunity is hindered by rapid therapeutic elimination of bacteria, but can be overcome by providing additional inflammatory and/or antigenic stimuli. PMID- 19542363 TI - Gene expression profiling and real-time PCR analyses identify novel potential cancer-testis antigens in multiple myeloma. AB - Cancer-testis (CT) Ags are attractive targets for immunotherapeutic strategies since they are aberrantly expressed in malignant cells and not, or in limited number, in somatic tissues, except germ cells. To identify novel CT genes in multiple myeloma, we used Affymetrix HG-U133 gene expression profiles of 5 testis, 64 primary multiple myeloma cells (MMC), and 24 normal tissue samples. A 5-filter method was developed to keep known CT genes while deleting non-CT genes. Starting from 44,928 probe sets, including probe sets for 18 previously described CT genes, we have obtained 82 genes expressed in MMC and testis and not detected in more than 6 normal tissue samples. This list includes 14 of the 18 known CT genes and 68 novel putative CT genes. Real-time RT-PCR was performed for 34 genes in 12 normal tissue samples, 5 MMC samples, and one sample of five pooled testes. It has validated the CT status of 23 of 34 genes (67%). We found one novel "testis-restricted" gene (TEX14, expression in testis and tumor only), eight "tissue-restricted" (mRNA detected in one or two nongametogenic tissues), and seven "differentially expressed" (mRNA detected in three to six nongametogenic tissues) CT genes. Further studies are warranted to determine the immunogenicity of these novel CT Ag candidates. PMID- 19542364 TI - Glycogen synthase kinase-3beta facilitates IFN-gamma-induced STAT1 activation by regulating Src homology-2 domain-containing phosphatase 2. AB - Glycogen synthase kinase-3beta (GSK-3beta)-modulated IFN-gamma-induced inflammation has been reported; however, the mechanism that activates GSK-3beta and the effects of activation remain unclear. Inhibiting GSK-3beta decreased IFN gamma-induced inflammation. IFN-gamma treatment rapidly activated GSK-3beta via neutral sphingomyelinase- and okadaic acid-sensitive phosphatase-regulated dephosphorylation at Ser(9), and proline-rich tyrosine kinase 2 (Pyk2)-regulated phosphorylation at Tyr(216). Pyk2 was activated through phosphatidylcholine specific phospholipase C (PC-PLC)-, protein kinase C (PKC)-, and Src-regulated pathways. The activation of PC-PLC, Pyk2, and GSK-3beta was potentially regulated by IFN-gamma receptor 2-associated Jak2, but it was independent of IFN-gamma receptor 1. Furthermore, Jak2/PC-PLC/PKC/cytosolic phospholipase A(2) positively regulated neutral sphingomyelinase. Inhibiting GSK-3beta activated Src homology-2 domain-containing phosphatase 2 (SHP2), thereby preventing STAT1 activation in the late stage of IFN-gamma stimulation. All these results showed that activated GSK-3beta synergistically affected IFN-gamma-induced STAT1 activation by inhibiting SHP2. PMID- 19542365 TI - SHIP regulates the reciprocal development of T regulatory and Th17 cells. AB - Maintaining an appropriate balance between subsets of CD4(+) Th and T regulatory cells (Tregs) is critical to maintain immune homeostasis and prevent autoimmunity. Through a common requirement for TGF-beta, the development of peripherally induced Tregs is intimately linked to that of Th17 cells, with the resulting lineages depending on the presence of proinflammatory cytokines such as IL-6. Currently very little is known about the molecular signaling pathways that control the development of Tregs vs Th17 cells. Reduced activity of the PI3K pathway is required for TGF-beta-mediated induction of Foxp3 expression and the suppressive activity of Tregs. To investigate how negative regulators of the PI3K pathway impact Treg development, we investigated whether SHIP, a lipid phosphatase that regulates PI3K activity, also plays a role in the development and function of Tregs. SHIP-deficient Tregs maintained suppressive capacity in vitro and in a T cell transfer model of colitis. Surprisingly, SHIP-deficient Th cells were significantly less able to cause colitis than were wild-type Th cells due to a profound deficiency in Th17 cell differentiation, both in vitro and in vivo. The inability of SHIP-deficient T cells to develop into Th17 cells was accompanied by decreased IL-6-stimulated phosphorylation of STAT3 and an increased capacity to differentiate into Treg cells under the influence of TGF beta and retinoic acid. These data indicate that SHIP is essential for normal Th17 cell development and that this lipid phosphatase plays a key role in the reciprocal regulation of Tregs and Th17 cells. PMID- 19542366 TI - Peripheral induction of tolerance by retinal antigen expression. AB - The contribution of peripheral expression of tissue-specific CNS Ags to the generation of tolerance is uncertain. To study this question, we examined mice transgenic (Tg) for expression of beta-galactosidase (beta gal) on the retinal photoreceptor cell arrestin promoter, in conjunction with TCR Tg mice producing CD4(+) T cells specific for beta gal (beta galTCR). Several strategies were used to test the hypothesis that betagal expressed in the retina supported thymus independent tolerance and regulatory T cell development. Retinal expression generated an immunoregulatory response that depressed development of immune responses to beta gal following systemic immunization with beta gal. This regulation was transferable to naive mice by CD3(+)4(+)25(+) T cells from naive retinal beta gal(+) donors. Experiments that removed the beta gal(+) retina by enucleation showed that subsequent development of a regulatory response was lost. Adoptive transfer of CD25(-) beta galTCR T cells into retinal beta gal Tg mice on the Rag(-/-) background led to regulatory activity that limited lymphopenia induced proliferation of beta galTCR T cells in mice with retinal expression of beta gal and inhibited the ear-swelling assay for delayed type hypersensitivity. These results show that retinal expression of very small amounts of a tissue specific Ag can generate tolerance that includes regulatory T cells. PMID- 19542367 TI - Prostaglandin E2 differentially modulates proinflammatory/prodestructive effects of TNF-alpha on synovial fibroblasts via specific E prostanoid receptors/cAMP. AB - The present study investigated the influence of PGE(2), E prostanoid (EP) receptors, and their signaling pathways on matrix metalloproteinase (MMP)-1 and IL-6 expression in synovial fibroblasts (SFs) from rheumatoid arthritis (RA) patients. RASFs expressed all four EP receptors, with selective induction of EP2 by TNF-alpha. TNF-alpha time-dependently increased intracellular cAMP/protein kinase A signaling (maximum, 6-12 h) and PGE(2) secretion (maximum, 24 h). PGE(2) and the EP2 agonists butaprost or ONO-AE1-259 ((16)-9-deoxy-9beta-chloro-15-deoxy 16-hydroxy-17,17-trimethylene-19,20-didehydro PGE(1)), in turn, induced a rapid, time-dependent (maximum, 15-30 min) increase of cAMP. Additionally, cyclooxygenase-2 inhibition by NS-398 (N-(2-cyclohexyloxy-4-nitrophenyl) methanesulfonamide) reduced the TNF-alpha-induced increase in IL-6 mRNA/protein, which was restored by stimulation with PGE(2) or EP2, EP3, and EP4 agonists. In contrast, TNF-alpha-induced MMP-1 secretion was not influenced by NS-398 and diminished by PGE(2) via EP2. Finally, 3-isobutyl-1-methylxanthine enhanced the effects of PGE(2) on MMP-1, but not on IL-6 mRNA. In conclusion, PGE(2) differentially affects TNF-alpha-induced mRNA expression of proinflammatory IL-6 and prodestructive MMP-1 regarding the usage of EP receptors and the dependency on cAMP. Although specific blockade of EP2 receptors is considered a promising therapeutic strategy in RA, opposite regulation of proinflammatory IL-6 and prodestructive MMP-1 by PGE(2) via EP2 may require more complex approaches to successfully inhibit the cyclooxygenase-1/2 cAMP axis. PMID- 19542368 TI - HSV-1-induced SOCS-1 expression in keratinocytes: use of a SOCS-1 antagonist to block a novel mechanism of viral immune evasion. AB - Keratinocytes are important for the acute phase of HSV-1 infection and subsequent persistence in sensory nervous tissue. In this study, we showed that keratinocytes (HEL-30) were refractory to IFN-gamma induction of an antiviral state to HSV-1 infection, while IFN-gamma did induce an antiviral state in fibroblasts (L929). This led us to examine the possible role of suppressor of cytokine signaling-1 (SOCS-1) in this refractiveness. RT-PCR analysis of SOCS-1 mRNA expression in HSV-1-infected cells showed a 4-fold increase for keratinocytes while having a negligible effect on fibroblasts. A similar pattern was observed at the level of SOCS-1 protein induction. Activation of STAT1alpha in keratinocytes was inhibited by HSV-1 infection. A direct effect of HSV-1 on the SOCS-1 promoter was shown in a luciferase reporter gene assay. We have developed a small peptide antagonist of SOCS-1, pJAK2(1001-1013), that had both an antiviral effect in keratinocytes against HSV-1 as well as a synergistic effect on IFN-gamma induction of an antiviral state. HSV-1 ICP0 mutant was inhibited by IFN-gamma in HEL-30 cells and was less effective than wild-type virus in induction of SOCS-1 promoter. We conclude that SOCS-1 plays an important role in the inhibition of the antiviral effect of IFN-gamma in keratinocytes infected with HSV-1. The use of SOCS-1 antagonist to abrogate this refractiveness could have a transformational effect on therapy against viral infections. PMID- 19542369 TI - The influence of IgE-enhancing and IgE-suppressive gammadelta T cells changes with exposure to inhaled ovalbumin. AB - It has been reported that the IgE response to allergens is influenced by gammadelta T cells. Intrigued by a study showing that airway challenge of mice with OVA induces in the spleen the development of gammadelta T cells that suppress the primary IgE response to i.p.-injected OVA-alum, we investigated the gammadelta T cells involved. We found that the induced IgE suppressors are contained within the Vgamma4(+) subset of gammadelta T cells of the spleen, that they express Vdelta5 and CD8, and that they depend on IFN-gamma for their function. However, we also found that normal nonchallenged mice harbor IgE enhancing gammadelta T cells, which are contained within the larger Vgamma1(+) subset of the spleen. In cell transfer experiments, airway challenge of the donors was required to induce the IgE suppressors among the Vgamma4(+) cells. Moreover, this challenge simultaneously turned off the IgE enhancers among the Vgamma1(+) cells. Thus, airway allergen challenge differentially affects two distinct subsets of gammadelta T cells with nonoverlapping functional potentials, and the outcome is IgE suppression. PMID- 19542370 TI - B cell intrinsic MyD88 signals drive IFN-gamma production from T cells and control switching to IgG2c. AB - The question of whether Ab responses to T-dependent Ags require B cell intrinsic signaling via the main TLR adaptor (MyD88) has become embroiled in confusion. In part this may be related to the methods used to analyze B cell intrinsic signaling. We have used a mixed bone marrow chimera model to generate mice in which the B cell compartment is completely deficient in MyD88 expression, while the other hematopoietic lineages are largely normal. These mice were immunized with T-dependent Ags or infected with Salmonella. We found that the Ag-specific IgG2c primary response was absolutely dependent on MyD88 signaling to B cells, while other Ig classes were not (IgG1 and IgG3) or much less so (IgG2b, IgA). The MyD88(B-/-) chimeric mice exhibited an impairment of development of IFN-gamma effector T cells, a likely contributory factor in the lack of IgG2c. We also found that B cell intrinsic MyD88 signals are required for the production of natural Abs. The data emphasize the nonredundant role of B cells as programmers of T cell differentiation in vivo. PMID- 19542371 TI - Tristetraprolin is required for full anti-inflammatory response of murine macrophages to IL-10. AB - IL-10 is essential for inhibiting chronic and acute inflammation by decreasing the amounts of proinflammatory cytokines made by activated macrophages. IL-10 controls proinflammatory cytokine and chemokine production indirectly via the transcription factor Stat3. One of the most physiologically significant IL-10 targets is TNF-alpha, a potent proinflammatory mediator that is the target for multiple anti-TNF-alpha clinical strategies in Crohn's disease and rheumatoid arthritis. The anti-inflammatory effects of IL-10 seem to be mediated by several incompletely understood transcriptional and posttranscriptional mechanisms. In this study, we show that in LPS-activated bone marrow-derived murine macrophages, IL-10 reduces the mRNA and protein levels of TNF-alpha and IL-1alpha in part through the RNA destabilizing factor tristetraprolin (TTP). TTP is known for its central role in destabilizing mRNA molecules containing class II AU-rich elements in 3' untranslated regions. We found that IL-10 initiates a Stat3-dependent increase of TTP expression accompanied by a delayed decrease of p38 MAPK activity. The reduction of p38 MAPK activity releases TTP from the p38 MAPK mediated inhibition, thereby resulting in diminished mRNA and protein levels of proinflammatory cytokines. These findings establish that TTP is required for full responses of bone marrow-derived murine macrophages to IL-10. PMID- 19542372 TI - Cutting edge: TNF-alpha mediates sensitization to ATP and silica via the NLRP3 inflammasome in the absence of microbial stimulation. AB - The Nlrp3 inflammasome is critical for the activation of caspase-1 in response to danger signals and particulate matter. However, its role in sterile inflammation remains unclear because prestimulation of phagocytic cells with microbial molecules is required for caspase-1 activation. We show here that exposure of macrophages and dendritic cells to TNF-alpha promotes ATP- or silica-mediated caspase-1 activation and IL-1beta secretion in the absence of microbial stimulation. The effect of TNF-alpha was abolished in macrophages deficient in TNF receptor I and II, Nlrp3, or ASC, whereas that induced by TLR ligands required MyD88/Trif. In addition to TNF-alpha, IL-1alpha and IL-1beta promoted caspase-1 activation via Nlrp3 in response to ATP. Remarkably, macrophages tolerized to TNF-alpha, but not to LPS, retained full sensitivity to ATP stimulation via Nlrp3. These results provide a mechanism by which danger signals and particulate matter mediate inflammation via the Nlrp3 inflammasome in the absence of microbial infection. PMID- 19542373 TI - The cytoplasmic tail of the T cell receptor CD3 epsilon subunit contains a phospholipid-binding motif that regulates T cell functions. AB - The CD3 epsilon subunit of the TCR complex contains two defined signaling domains, a proline-rich sequence and an ITAM. We identified a third signaling sequence in CD3 epsilon, termed the basic-rich stretch (BRS). Herein, we show that the positively charged residues of the BRS enable this region of CD3 epsilon to complex a subset of acidic phospholipids, including PI(3)P, PI(4)P, PI(5)P, PI(3,4,5)P(3), and PI(4,5)P(2). Transgenic mice containing mutations of the BRS exhibited varying developmental defects, ranging from reduced thymic cellularity to a complete block in T cell development. Peripheral T cells from BRS-modified mice also exhibited several defects, including decreased TCR surface expression, reduced TCR-mediated signaling responses to agonist peptide-loaded APCs, and delayed CD3 epsilon localization to the immunological synapse. Overall, these findings demonstrate a functional role for the CD3 epsilon lipid-binding domain in T cell biology. PMID- 19542374 TI - A MyD88-dependent early IL-17 production protects mice against airway infection with the obligate intracellular pathogen Chlamydia muridarum. AB - We found that IL-17, a signature cytokine of Th17, was produced early in the innate immunity phase after an intranasal infection with the obligate intracellular pathogen Chlamydia muridarum. The airway IL-17, which peaked at 48 h after infection, was dependent on live chlamydial organism replication and MyD88-mediated signaling pathways. Treatment with antibiotics or knockout of the MyD88 gene, but not Toll/IL receptor domain-containing adapter-inducing IFN-beta, can block the early IL-17 production. Treatment of mice with an anti-IL-17 neutralizing mAb enhanced growth of chlamydial organisms in the lung, dissemination to other organs, and decreased mouse survival, whereas treatment with an isotype-matched control IgG had no effect. Although IL-17 did not directly affect chlamydial growth in cell culture, it enhanced the production of other inflammatory cytokines and chemokines by Chlamydia-infected cells and promoted neutrophil infiltration in mouse airways during chlamydial infection, which may contribute to the antichlamydial effect of IL-17. These observations suggest that an early IL-17 response as an innate immunity component plays an important role in initiating host defense against infection with intracellular bacterial pathogens in the airway. PMID- 19542375 TI - Prednisolone treatment induces tolerogenic dendritic cells and a regulatory milieu in myasthenia gravis patients. AB - FOXP3-expressing naturally occurring CD4(+)CD25(high) T regulatory cells (Treg) are relevant in the control of autoimmunity, and a defect in this cell population has been observed in several human autoimmune diseases. We hypothesized that altered functions of peripheral Treg cells might play a role in the immunopathogenesis of myasthenia gravis, a T cell-dependent autoimmune disease characterized by the presence of pathogenic autoantibodies specific for the nicotinic acetylcholine receptor. We report in this study a significant decrease in the in vitro suppressive function of peripheral Treg cells isolated from myasthenia patients in comparison to those from healthy donors. Interestingly, Treg cells from prednisolone-treated myasthenia gravis patients showed an improved suppressive function compared with untreated patients, suggesting that prednisolone may play a role in the control of the peripheral regulatory network. Indeed, prednisolone treatment prevents LPS-induced maturation of monocyte derived dendritic cells by hampering the up-regulation of costimulatory molecules and by limiting secretion of IL-12 and IL-23, and enhancing IL-10. In addition, CD4(+) T cells cultured in the presence of such tolerogenic dendritic cells are hyporesponsive and can suppress autologous CD4(+) T cell proliferation. The results shown in this study indicate that prednisolone treatment promotes an environment that favors immune regulation rather than inflammation. PMID- 19542377 TI - Thickening of small bowel folds. PMID- 19542376 TI - IL-17A controls IL-17F production and maintains blood neutrophil counts in mice. AB - G-CSF, its receptor, and IL-17 receptor A (IL-17RA) are all required to maintain baseline neutrophil counts in mice. In this study, we tested whether IL-17F could compensate and maintain baseline neutrophil counts in the absence of IL-17A. Unlike the reduced neutrophil counts found in IL-17RA-deficient mice, neutrophil counts were mildly increased in IL-17A-deficient (Il17a(-/-)) animals. There was no evidence for infection or altered neutrophil function. Plasma G-CSF and IL-17F levels were elevated in Il17a(-/-) compared with wild-type mice. IL-17F was mainly produced in the spleen and mesenteric lymph nodes, but IL-23 was unaltered in Il17a(-/-) mice. Instead, Il17a(-/-) splenocytes differentiated with IL-6, TGF beta, and IL-23 ex vivo produced significantly more IL-17F in response to IL-23 than wild-type cells. Adding rIL-17A to Il17a(-/-) splenocyte cultures reduced IL 17F mRNA and protein secretion. These effects were also observed in wild-type but not IL-17RA-deficient cells. We conclude that IL-17A mediated suppression of IL 17F production and secretion requires IL-17RA and is relevant to maintain the normal set point of blood neutrophil counts in vivo. PMID- 19542378 TI - Novel treatment options for chronic heart failure: a radiologist's perspective. AB - OBJECTIVE: This article aims to familiarize radiologists with novel treatment options for chronic heart failure that is unresponsive to medical therapy, such as mechanical cardiac assist devices, surgical procedures, resynchronization therapy with biventricular pacing, and cellular cardiomyoplasty, and their radiographic appearances. CONCLUSION: Heart transplantation as a treatment of debilitating heart failure provides an opportunity for meaningful long-term survival but is limited by a shortage of donor hearts. This has spurred the development of new treatment options for chronic heart failure that is unresponsive to medical therapy. PMID- 19542379 TI - Cardiac MRI for detection of unrecognized myocardial infarction in patients with end-stage renal disease: comparison with ECG and scintigraphy. AB - OBJECTIVE: The purposes of this study were to use the myocardial delayed enhancement technique of cardiac MRI to investigate the frequency of unrecognized myocardial infarction (MI) in patients with end-stage renal disease, to compare the findings with those of ECG and SPECT, and to examine factors that may influence the utility of these methods in the detection of MI. SUBJECTS AND METHODS: We prospectively performed cardiac MRI, ECG, and SPECT to detect unrecognized MI in 72 patients with end-stage renal disease at high risk of coronary artery disease but without a clinical history of MI. RESULTS: Fifty-six patients (78%) were men (mean age, 56.2 +/- 9.4 years) and 16 (22%) were women (mean age, 55.8 +/- 11.4). The mean left ventricular mass index was 103.4 +/- 27.3 g/m(2), and the mean ejection fraction was 60.6% +/- 15.5%. Myocardial delayed enhancement imaging depicted unrecognized MI in 18 patients (25%). ECG findings were abnormal in five patients (7%), and SPECT findings were abnormal in 19 patients (26%). ECG findings were false-negative in 14 cases and false positive in one case. The accuracy, sensitivity, and specificity of ECG were 79.2%, 22.2%, and 98.1% (p = 0.002). SPECT findings were false-negative in six cases and false-positive in seven cases. The accuracy, sensitivity, and specificity of SPECT were 81.9%, 66.7%, and 87.0% (not significant). During a period of 4.9-77.9 months, 19 cardiac deaths were documented, but no statistical significance was found in survival analysis. CONCLUSION: Cardiac MRI with myocardial delayed enhancement can depict unrecognized MI in patients with end stage renal disease. ECG and SPECT had low sensitivity in detection of MI. Infarct size and left ventricular mass can influence the utility of these methods in the detection of MI. PMID- 19542380 TI - Defining normal vertebral angulation at the thoracolumbar junction. AB - OBJECTIVE: The purpose of this cross-sectional study is to define the normal range of endplate angulation at T12 and L1 and, by doing so, to validate the angle measurement tools that are readily available on nearly all PACS. MATERIALS AND METHODS: Two hundred consecutive lateral scout CT scans were examined in patients who were either 25 (n = 100) or 35 (n = 100) years old. The endplate angles for T12 and L1 were measured using a "Cobb angle" tool on a standard PACS workstation. Twenty-two cadaveric vertebrae were also imaged, and measurements obtained from the lateral scout CT image using electronic calipers were compared with measurements obtained with a goniometer. RESULTS: The mean endplate angle at T12 measures 4.34 degrees (2 SD, 4.5 degrees) and at L1, 4.48 degrees (4.26 degrees). The normal range of endplate angulation is therefore -0.16 degrees to 8.84 degrees at T12 and 0.22-8.74 degrees at L1. No statistically significant difference was seen in the endplate angulation when men were compared with women or 25- and 35-year-old age groups were compared. A strong correlation exists between direct and CT-derived endplate angle measurements. CONCLUSION: Vertebral endplate angulation can be reliably measured using widely available PACS workstation tools. The mean endplate angle for T12 and L1 is approximately 4.5 degrees, with an approximate range extending from 0 degrees to 9 degrees. For practical purposes, an endplate angle of 10 degrees or more can be considered outside the normal range. PMID- 19542381 TI - Palliative treatment of inoperable malignant tracheobronchial obstruction: temporary stenting combined with radiation therapy and/or chemotherapy. AB - OBJECTIVE: The purpose of our study was to evaluate the clinical results of temporary stenting followed by radiation and/or chemotherapy in patients with inoperable malignant tracheobronchial strictures. CONCLUSION: Temporary stenting combined with radiation therapy and/or chemotherapy may be clinically effective in the palliative treatment of patients with malignant tracheobronchial strictures. Stent placement may serve as an effective bridge to radiation and/or chemotherapy, allowing the latter to consolidate durable relief of obstructing symptoms by reducing tumor burden. Furthermore, our treatment strategy may increase patients' quality of life by reducing stent-related complications. PMID- 19542383 TI - CT angiography signs of lower extremity vascular trauma. AB - OBJECTIVE: Specific CT angiography (CTA) signs of vascular injury can be readily detected, and additional information regarding osseous and soft-tissue injuries can also be routinely obtained. In this article, we illustrate the important CTA signs of lower extremity vascular injury. CONCLUSION: CTA is efficient and accurate in the evaluation of clinically significant lower extremity arterial injuries after trauma. PMID- 19542382 TI - Risk factors involved in the development of pneumothorax during radiofrequency ablation of lung neoplasms. AB - OBJECTIVE: The purpose of this study was to retrospectively evaluate the risk factors involved in the development of pneumothorax during radiofrequency ablation of lung tumors. MATERIALS AND METHODS: This retrospective study covered 124 ablation sessions for lung tumors (10 primary lesions, 114 metastatic lesions) in 82 patients (46 men, 36 women; mean age, 64.0 years) treated between December 2005 and January 2008. The exclusion criteria for ablation therapy were lesions with a maximal diameter greater than 5 cm and the presence of more than five lesions. A bipolar electrode needle was used under CT guidance. Four patients were treated with two ablation electrodes simultaneously. RESULTS: The incidence of pneumothorax (detected with CT) was 11.3% (14 of 124 sessions). Pneumothorax was graded mild (lung surface retraction, < or = 2 cm), moderate (lung surface retraction, 2-4 cm), or severe (lung surface retraction, > or = 4 cm). Significant risk factors encountered in the development of pneumothorax were age greater than 60 years (p = 0.046), emphysema (p = 0.02), tumor diameter < or = 1.5 cm (p = 0.0008), lesions in lower part of lung, (p = 0.027), aerated lung parenchyma traversed by the needle track for a distance > or = 2.6 cm (p = 0.0017), and traversal of a major pulmonary fissure (p = 0.0004). Pneumothorax developed in one of the four patients in whom multiple electrodes were used. The mean depth of lung lesions complicated by pneumothorax was 2.9 +/- 1.55 cm (range, 0-5.5 cm). Conservative treatment was performed in four of the 14 pneumothorax sessions (28.6%). In six of the 14 sessions (42.9%), immediate complete evacuation was achieved with an intercostal catheter and manual evacuation; chest tube placement was indicated in four sessions (28.6%). Two patients were treated with manual evacuation because evidence of a progressive increase in pneumothorax on the 24-hour follow-up CT scan indicated failure of conservative treatment. CONCLUSION: The development of pneumothorax complicating radiofrequency ablation can be unpredictable, but the many risk factors involved can make the incidence higher among some patients than others. Some of these risk factors are technically avoidable and have to be ruled out. PMID- 19542384 TI - Invasive micropapillary carcinoma of the breast: mammographic, sonographic, and MRI features. AB - OBJECTIVE: The purpose of this study was to describe the clinical, imaging, and histopathologic findings of invasive micropapillary carcinoma of the breast. CONCLUSION: The imaging characteristics of invasive micropapillary carcinoma are highly suggestive of malignancy. The lesion is a high-density irregular mass with indistinct margins associated with microcalcifications on mammograms; a solid irregular hypoechoic mass with indistinct margins and frequent axillary nodal involvement on sonograms; and a multifocal mass on MR images. This tumor may necessitate aggressive management. PMID- 19542385 TI - Dermatofibrosarcoma protuberans of the breast: imaging features and review of the literature. AB - OBJECTIVE: The objective of our study was to present the imaging features, including an MRI example, of dermatofibrosarcoma protuberans of the breast, an uncommon soft-tissue neoplasm of the breast, and review the literature. CONCLUSION: Dermatofibrosarcoma protuberans is an extremely rare malignancy of the breast, with few published reports. This is the largest collection of such cases in a single institution with analysis of the imaging features. PMID- 19542386 TI - Hepatocellular carcinoma variants: radiologic-pathologic correlation. AB - OBJECTIVE: The purpose of this article is to show the imaging findings of variant types of hepatocellular carcinoma (HCC) with pathologic correlations. CONCLUSION: The variant types of HCC may not share its typical imaging characteristics. An understanding of the radiologic findings for variant types of HCC can be helpful in the differential diagnosis of hepatic tumors. PMID- 19542387 TI - Invasive carcinoma of the breast accompanied by coarse calcification. AB - OBJECTIVE: We describe the case of a woman with a left breast mass. At mammography, the mass was shown to be irregular and accompanied by coarse calcification. Core needle biopsy revealed invasive carcinoma and a mastectomy was performed. Histopathology showed fibrosis with partial hyalinization eccentrically placed within the tumor with a large area of calcification at the core. CONCLUSION: Benign calcifications within a breast mass are not diagnostic of a benign process if the imaging characteristics of the mass are suspicious. PMID- 19542389 TI - Prone breast dual-time-point PET/CT compared with MRI for determining breast cancer. PMID- 19542388 TI - Humeral avulsion of the glenohumeral ligament. PMID- 19542390 TI - CT of rib lesions. AB - OBJECTIVE: This article discusses how ribs are involved in a variety of traumatic, metabolic, inflammatory, neoplastic, and congenital disorders. CONCLUSION: We review the normal anatomy pertinent to rib imaging and illustrate the key features of a variety of rib lesions, emphasizing the diagnostic value of CT. PMID- 19542392 TI - In defense of body CT. AB - OBJECTIVE: Rapid technical developments and an expanding list of applications that have supplanted less accurate or more invasive diagnostic tests have led to a dramatic increase in the use of body CT in medical practice since its introduction in 1975. Our purpose here is to discuss medical justification of the small potential risk associated with the ionizing radiation used in CT and to provide perspectives on practice-specific decisions that can maximize overall patient benefit. In addition, we review available dose management and optimization techniques. CONCLUSION: Dose reduction strategies described in this article must be well understood and properly used, but also require broad-based practice strategies that extend beyond the CT scanner console and default, generic manufacturer settings. In the final analysis, physicians must request the imaging examination that best addresses the specific medical question without allowing worries about radiation to dissuade them or their patients from obtaining needed CT examinations. Ongoing efforts to ensure that CT examinations are both medically justified and optimally performed must continue, and education must be provided to the medical community and general public that put both the potential risks--and benefits--of CT examinations into proper perspective. PMID- 19542393 TI - Colorectal cancer screening with CT colonography: key concepts regarding polyp prevalence, size, histology, morphology, and natural history. AB - OBJECTIVE: The purpose of this article is to provide a timely update on a variety of key polyp topics to construct a proper framework for physicians who are interested in providing CT colonography screening as a clinical service. CONCLUSION: As the medical community considers the expansion of CT colonography for screening, we believe it is prudent to update and review several key concepts regarding colorectal polyps. In particular, it is important to replace the older literature derived from high-risk and symptomatic cohorts with the wealth of newer and more applicable data from average-risk and asymptomatic screening cohorts. Familiarity with current concepts regarding flat (nonpolypoid) lesions and the natural history of small colorectal polyps is also vital to the effective application of this technique. PMID- 19542391 TI - Advanced MRI methods for assessment of chronic liver disease. AB - OBJECTIVE: With recent advances in technology, advanced MRI methods such as diffusion-weighted and perfusion-weighted MRI, MR elastography, chemical shift based fat-water separation, and MR spectroscopy can now be applied to liver imaging. We will review the respective roles of these techniques for assessment of chronic liver disease. CONCLUSION: MRI plays an increasingly important role in assessment of patients with chronic liver disease because of the lack of ionizing radiation and the possibility of performing multiparametric imaging. PMID- 19542394 TI - Dual-energy and low-kVp CT in the abdomen. AB - OBJECTIVE: The purpose of this article is to discuss the influence of tube potential on CT images and explore the potential impact of dual-energy CT on imaging of the abdomen and pelvis. CONCLUSION: Low peak tube voltage (kVp) settings provide high conspicuity of contrast materials at CT but may result in high image noise, particularly in larger patients. Material decomposition at dual energy CT can differentiate renal stones by their composition, quantify tissue iron stores, improve the detection of pathologic hyperenhancement, and reduce contrast material and radiation dose compared with conventional CT. Further clinical research and technique refinement will be needed as the usage of these exciting technologies spreads. PMID- 19542395 TI - Contrast-enhanced ultrasound: what is the evidence and what are the obstacles? AB - OBJECTIVE: Although ultrasound contrast agents (UCAs) are popular and widely used in Europe and Asia, the U.S. Food and Drug Administration (FDA) has not approved a microbubble agent for radiology imaging in the United States. Herein, we discuss the evidence for and the obstacles to using UCAs for contrast-enhanced ultrasound (CEUS). CONCLUSION: Despite the obstacles to the use of UCAs for CEUS including regulatory and practice patterns, the evidence indicates that radiologists and patients will be missing an effectual imaging option if we do not encourage the use of CEUS and strongly support the approval of UCAs by the FDA. The evidence outweighs the obstacles: CEUS is cost-effective; can be performed at the bedside; uses no ionizing radiation; has no nephrotoxicity; and, most importantly, can provide accurate diagnostic information comparable to CT and MRI. PMID- 19542396 TI - Features of nephrogenic systemic fibrosis on radiology examinations. AB - OBJECTIVE: The objective of this article is to illustrate the spectrum of imaging findings with photographic and histopathologic correlation in patients with biopsy-proven nephrogenic systemic fibrosis (NSF). CONCLUSION: Features of NSF may be evident on the patient's skin as well as on routine imaging studies, although these imaging findings are nonspecific and are more likely to occur with other diseases. PMID- 19542397 TI - Sensitivity of CT colonography for nonpolypoid colorectal lesions interpreted by human readers and with computer-aided detection. AB - OBJECTIVE: The purpose of our study was to determine the sensitivity of CT colonography (CTC) interpreted by human readers and with computer-aided detection (CAD) for genuinely nonpolypoid colorectal lesions, defined as 2 mm or less in lesion height at colonoscopy. MATERIALS AND METHODS: A computerized database search for a 33-month period found 21 patients who had undergone both colonoscopy and CTC and who had a total of 23 genuinely nonpolypoid colorectal lesions: eight adenomas (9-30 mm in width), 10 stage Tis or T1 adenocarcinomas (10-25 mm), and five nonadenomatous lesions (8-20 mm). CTC was performed using a cathartic preparation and fecal tagging and was interpreted by experienced readers in a blinded manner using a primary 3D method and with CAD. RESULTS: The sensitivities of human readers for nonpolypoid adenomatous lesions (i.e., both adenomas and adenocarcinomas), adenocarcinomas, and nonadenomatous lesions were 66.7% (12/18), 90% (9/10), and 0% (0/5), respectively. Sensitivities were 55.6% (10/18), 90% (9/10), and 0% (0/5) for CAD. A 10-mm stage T1 adenocarcinoma was missed by a human reader on blinded review but was detected with CAD. Both human readers and CAD yielded significantly higher sensitivity for adenomatous lesions than for nonadenomatous lesions (p = 0.014 and 0.046, respectively) and for adenocarcinomas than for noncancerous lesions (p = 0.003 and 0.0001, respectively). CONCLUSION: CTC showed a high sensitivity for nonpolypoid stage Tis and T1 adenocarcinomas 10 mm or greater in width despite the limited overall sensitivity for nonpolypoid adenomatous lesions, when performed using cathartic preparation and fecal tagging. PMID- 19542398 TI - Volume imaging in the abdomen with ultrasound: how we do it. AB - OBJECTIVE: The objective of our study was to evaluate the feasibility of volumetric acquisition of the abdominal organs using performance guidelines that we developed in our preliminary experience. MATERIALS AND METHODS: Mechanical volumetric acquisitions of each abdominal organ, including the liver, gallbladder, pancreas, kidneys, spleen, bowel, and aorta, were performed in 200 consecutive patients. RESULTS: One thousand four hundred fifty-four volume data sets were graded for feasibility of performance and technical adequacy from I (impossible, incomplete) to V (excellent, complete). The most successfully imaged organ was the right kidney (grades IV and V, 95.0%) and the least successfully imaged, the spleen (grades IV and V, 69.0%). Very good to excellent grades (IV and V) were obtained in 1,215 (83.6%) of the 1,454 volumes. One hundred twelve (7.7%) of the 1,454 volumes were failures (grades I and II). The three organs with the highest success compared with the right kidney were the left kidney, gallbladder, and liver. The data sets of all the other organs showed a statistically significant difference in the feasibility of performance from the right kidney. Liver acquisition failures were associated with end-stage liver cirrhosis (n = 6), fatty liver (n = 3), and obesity (n = 3). Other acquisition failures, similar to conventional sonography, were associated with bowel gas interference and poor acoustic window. The technical limitations include poor resolution in the B and C planes and a limited range of frequencies; these limitations can be overcome in the future with matrix transducers and introduction of the technology to a broader frequency range. CONCLUSION: Volumetric acquisition in the abdomen performed using defined guidelines is feasible with recognized limitations. Technology advances will improve this imaging technique in the future. PMID- 19542399 TI - Efficacy of perflubutane microbubble-enhanced ultrasound in the characterization and detection of focal liver lesions: phase 3 multicenter clinical trial. AB - OBJECTIVE: The purpose of this study was to assess the efficacy and safety of contrast-enhanced ultrasound performed with perflubutane microbubbles in comparison with unenhanced ultrasound and dynamic CT in the characterization of focal liver lesions during the vascular phase of imaging and in the detection of lesions during the Kupffer phase. SUBJECTS AND METHODS: A total of 196 patients were enrolled at 15 centers in Japan. Vascular phase images were obtained before contrast injection until 1 minute after injection. Kupffer phase images were obtained 10 minutes after injection. Dual-phase CT was performed as determined by standard clinical practice at each center. Unenhanced ultrasound, contrast enhanced ultrasound, and CT images were read by blinded reviewers, and the results they reached regarding characterization and detection were compared with reference standard findings made by onsite investigators. The safety observation period was 72 hours after contrast administration. RESULTS: Among the 190 patients included in the characterization analysis, the accuracy of contrast enhanced ultrasound (88.9%) was significantly greater than that of unenhanced ultrasound (68.4%) and dynamic CT (80.5%) (p < 0.001 and p = 0.008). Among the 191 patients in the detection analysis, the efficacy of contrast-enhanced ultrasound in detection of lesions was significantly higher than that of unenhanced ultrasound and dynamic CT (p < 0.001 and p = 0.008), predominantly because more metastatic lesions were detected (both p < 0.001). In particular, contrast-enhanced ultrasound was superior to dynamic CT in the detection of metastatic lesions measuring 1 cm or smaller. The incidence of adverse events was 49.2% and that of adverse drug reactions was 10.4%. All adverse drug reactions were mild. CONCLUSION: Compared with unenhanced ultrasound and dynamic CT, contrast-enhanced ultrasound with perflubutane microbubbles improved diagnostic efficacy in both characterization and detection of focal liver lesions with no serious adverse drug reactions. PMID- 19542400 TI - Diagnosis of acute appendicitis with sliding slab ray-sum interpretation of low dose unenhanced CT and standard-dose i.v. contrast-enhanced CT scans. AB - OBJECTIVE: The purpose of this study was to compare low-dose unenhanced CT with standard-dose i.v. contrast-enhanced CT in the diagnosis of appendicitis. MATERIALS AND METHODS: Two hundred seven adults with suspected appendicitis underwent CT with mean effective doses of both 4.2 and 8.0 mSv. Two radiologists retrospectively reviewed thin-section images by sliding a 5-mm-thick ray-sum slab. They rated the likelihood of appendicitis and appendiceal visualization on 5- and 3-point scales, respectively, and proposed alternative diagnoses. Likelihood > or = 3 was considered a positive diagnosis. Receiver operating characteristics analysis, the McNemar test, and the Wilcoxon's signed-rank test were used. RESULTS: Seventy-eight patients had appendicitis. The values of the area under the receiver operating characteristics curve were 0.98 for the low dose unenhanced acquisition and 0.97 for the standard-dose contrast-enhanced acquisition for reader 1 (95% CI for the difference, -0.02 to 0.03) and 0.99 and 0.98 (-0.02 to 0.02) for reader 2. Sensitivity was 98.7% for low-dose unenhanced CT and 100% for standard-dose contrast-enhanced CT for reader 1 (p = 1.00) and 100% for both techniques for reader 2. Specificity was 95.3% and 93.0% (p = 0.25) and 96.9% and 96.9%. The interpretation was indeterminate (score 3) in 0.5% and 1.4% of cases for reader 1 (p = 0.63) and 0.5% and 0% for reader 2 (p = 1.00). A normal appendix was not visualized in 5.4% and 3.9% of cases by reader 1 (p = 0.63) and 3.9% and 2.3% of cases by reader 2 (p = 0.50). None of the patients whose appendix was not visualized had appendicitis. Diagnostic confidence, visualization score for a normal appendix, and correct alternative diagnosis tended to be compromised with use of low-dose unenhanced CT, showing a significant difference for a reader's confidence in the diagnosis of appendicitis (p = 0.004). The two techniques were comparable in the diagnosis of appendiceal perforation. CONCLUSION: Low-dose unenhanced CT is potentially useful in the diagnosis of appendicitis. PMID- 19542401 TI - Unenhanced MR portography with a half-Fourier fast spin-echo sequence and time space labeling inversion pulses: preliminary results. AB - OBJECTIVE: For this study, we aimed to selectively visualize the intrahepatic portal veins using 3D half-Fourier fast spin-echo (FSE) MR angiography (MRA) with a time-space labeling inversion pulse (T-SLIP) and to optimize the acquisition protocol. SUBJECTS AND METHODS: Respiratory-triggered 3D half-Fourier FSE scans were obtained in 25 healthy adult subjects combined with two different T-SLIPs: one placed on the liver and the thorax to suppress signals of the liver parenchyma, hepatic veins, and abdominal arteries and the other on the lower abdomen to suppress the ascending signal of the inferior vena cava. One of the most important factors was the inversion time (TI) of the inversion pulse for the liver and thorax. Image quality was evaluated in terms of signal-to-noise ratio, contrast-to-noise ratio, and mean visualization scores at four different TIs: 800, 1,200, 1,600, and 2,000 milliseconds. RESULTS: Selective visualization of the portal vein was successfully achieved in all volunteers, and anatomic variations were also seen in three subjects. A TI of 1,200 milliseconds was optimal in our protocol because it was sufficient for peripheral portal vein visualization and was most suitable for signal suppression of the hepatic veins and liver parenchyma. CONCLUSION: Half-Fourier FSE scanning with T-SLIPs enabled selective visualization of the portal vein without an exogenous contrast agent. PMID- 19542402 TI - Prospective comparison of state-of-the-art MR enterography and CT enterography in small-bowel Crohn's disease. AB - OBJECTIVE: The objective of our study was to prospectively obtain pilot data on the accuracy of MR enterography for detecting small-bowel Crohn's disease compared with CT enterography and with a clinical reference standard based on imaging, clinical information, and ileocolonoscopy. SUBJECTS AND METHODS: The study group for this blinded prospective study was composed of 33 patients with suspected active Crohn's ileal inflammation who were scheduled for clinical CT enterography and ileocolonoscopy and had consented to also undergo MR enterography. The MR enterography and CT enterography examinations were each interpreted by two radiologists with disagreements resolved by consensus. The reports from ileocolonoscopy with or without mucosal biopsy were interpreted by a gastroenterologist. The reference standard for the presence of small-bowel Crohn's disease was based on the final clinical diagnosis by the referring gastroenterologist after reviewing all of the available information. RESULTS: All 33 patients underwent CT enterography and ileocolonoscopy, 30 of whom also underwent MR enterography. The sensitivities of MR enterography and CT enterography for detecting active small-bowel Crohn's disease were similar (90.5% vs 95.2%, respectively; p = 0.32). The image quality scores for MR enterography examinations were significantly lower than those for CT enterography (p = 0.005). MR enterography and CT enterography identified eight cases (24%) with a final diagnosis of active small-bowel inflammation in which the ileal mucosa appeared normal at ileocolonoscopy. Furthermore, enterography provided the only available imaging in three additional patients who did not have ileal intubation. CONCLUSION: MR enterography and CT enterography have similar sensitivities for detecting active small-bowel inflammation, but image quality across the study cohort was better with CT. Cross-sectional enterography provides complementary information to ileocolonoscopy. PMID- 19542404 TI - Doppler ultrasound findings in the hepatic artery shortly after liver transplantation. AB - OBJECTIVE: The purpose of this article is to describe the Doppler waveform findings in the hepatic artery in the early posttransplantation period, both in the absence and presence of arterial complications. CONCLUSION: The presence of transient high-resistance Doppler waveforms in normal hepatic arteries is a common finding after grafting. Hepatic artery thrombosis and stenosis, and arterial steal syndromes can be diagnosed by Doppler in the early liver transplantation period. PMID- 19542403 TI - Feasibility of in vivo MR elastographic splenic stiffness measurements in the assessment of portal hypertension. AB - OBJECTIVE: Liver stiffness is associated with portal hypertension in patients with chronic liver disease. However, the relation between spleen stiffness and clinically significant portal hypertension remains unknown. The purposes of this study were to determine the feasibility of measuring spleen stiffness with MR elastography and to prospectively test the technique in healthy volunteers and in patients with compensated liver disease. MATERIALS AND METHODS: Spleen stiffness was measured with MR elastography in 12 healthy volunteers (mean age, 37 years; range, 25-82 years) and 38 patients (mean age, 56 years; range, 36-60 years) with chronic liver disease of various causes. For patients with liver disease, laboratory findings, spleen size, presence and size of esophageal varices, and liver histologic results were recorded. Statistical analyses were performed to assess all measurements. RESULTS: MR elastography of the spleen was successfully performed on all volunteers and patients. The mean spleen stiffness was significantly lower in the volunteers (mean, 3.6 +/- 0.3 kPa) than in the patients with liver fibrosis (mean, 5.6 +/- 5.0 kPa; range, 2.7-19.2 kPa; p < 0.001). In addition, a significant correlation was observed between liver stiffness and spleen stiffness for the entire cohort (r(2) = 0.75; p < 0.001). Predictors of spleen stiffness were splenomegaly, spleen volume, and platelet count. A mean spleen stiffness of 10.5 kPa or greater was identified in all patients with esophageal varices. CONCLUSION: MR elastography of the spleen is feasible and shows promise as a quantitative method for predicting the presence of esophageal varices in patients with advanced hepatic fibrosis. PMID- 19542405 TI - Typical and atypical manifestations of serous cystadenoma of the pancreas: imaging findings with pathologic correlation. AB - OBJECTIVE: The purpose of this article is to present the typical and atypical manifestations of serous cystadenoma, which can be visualized with cross sectional imaging. CONCLUSION: Serous cystadenomas of the pancreas have various distinguishing imaging features. Typically, a serous cystadenoma is morphologically classified as having either a polycystic, honeycomb, or oligocystic pattern. Atypical manifestations of serous cystadenoma can include giant tumors with ductal dilatation, intratumoral hemorrhages, solid variants, unilocular cystic tumors, interval growth, and a disseminated form. PMID- 19542406 TI - Dual-energy CT for the assessment of contrast material distribution in the pulmonary parenchyma. AB - OBJECTIVE: The purpose of this study was to assess the feasibility and diagnostic value of dual-energy CT iodine mapping at pulmonary CT angiography. SUBJECTS AND METHODS: Ninety-three patients underwent CT angiography with the dual-energy technique on a dual-source CT scanner. Postprocessing was used to map iodine in the lung parenchyma on the basis of its spectral behavior, and image quality was assessed by two readers. Iodine distribution patterns were rated as homogeneous, patchy, or circumscribed defects. Conventional CT angiographic images reconstructed from the same data sets were reviewed for the presence and localization of pulmonary embolism, whether embolic occlusion was partial or complete, and the presence of changes in the lung parenchyma. Dual-energy perfusion findings were correlated with the CT angiographic and lung-window CT findings in per-patient and per-segment analyses. RESULTS: Iodine distribution was homogeneous in 49 patients, of whom CT angiography showed no pulmonary embolism in 46 patients and nonocclusive pulmonary emboli in three patients. Images of 29 patients showed a patchy pattern; 24 of these patients had no pulmonary embolism, and five had nonocclusive pulmonary emboli with solely nonocclusive intravascular clots. Images of 15 patients showed segmental or subsegmental defects; four of these patients had evidence of pulmonary embolism, and 11 had occlusive pulmonary emboli with at least one occlusive clot in the pulmonary vasculature. CONCLUSION: Dual-energy CT is reliable in the detection of defects in pulmonary parenchymal iodine distribution that correspond to embolic vessel occlusion. PMID- 19542407 TI - Low-risk patients with chest pain in the emergency department: negative 64-MDCT coronary angiography may reduce length of stay and hospital charges. AB - OBJECTIVE: The current standard-of-care workup of low-risk patients with chest pain in an emergency department takes 12-36 hours and is expensive. We hypothesized that negative 64-MDCT coronary angiography early in the workup of such patients may enable a shorter length of stay and reduce charges. MATERIALS AND METHODS: The standard-of-care evaluation consisted of serial cardiac enzyme tests, ECGs, and stress testing. After informed consent, we added cardiac CT early in the standard-of-care workup of 53 consecutive patients. Fifty patients had negative CT findings and were included in this series. The length of stay and charges were analyzed using actual patient data for all patients in the standard of-care workup and for two earlier discharge scenarios based on negative cardiac CT results: First, CT plus serial enzyme tests and ECGs during an observation period followed by discharge if all were negative; and second, CT plus one set of enzyme tests and one ECG followed by discharge if all were negative. Comparisons were made using paired Student's t tests. RESULTS: For standard of care and the two CT-based earlier discharge analyses, the mean lengths of stay were 25.4, 14.3, and 5.0 hours; mean charges were $7,597, $6,153, and $4,251. Length of stay and charges were both significantly less (p < 0.001) for the two CT-based analyses. CONCLUSION: In low-risk patients with chest pain, discharge from the emergency department based on negative cardiac CT, enzyme tests, and ECG may significantly decrease both length of stay and hospital charges compared with the standard of care. PMID- 19542408 TI - Best ways to provide feedback to radiologists on mammography performance. AB - OBJECTIVE: The Mammography Quality Standards Act of 1992 required a minimum performance audit of radiologists performing mammography. Since then, no studies have evaluated radiologists' perceptions of their audit reports, such as which performance measures are the most or least useful, or what the best formats are to present performance data. MATERIALS AND METHODS: We conducted a qualitative study with focus groups and interviews of 25 radiologists currently practicing mammography. All radiologists practiced at one of three sites in the Breast Cancer Surveillance Consortium (BCSC). The discussion guide included open-ended questions to elicit opinions on the following subjects: the most useful performance outcome measures, examples of reports and formats that are easiest to understand (e.g., graphs or tables), thoughts about comparisons between individual-level and aggregate data, and ideas about additional performance measures they would find useful. All discussions were tape-recorded and transcribed. We developed a set of themes and used ethnographic software to qualitatively analyze and extract quotes from transcripts. RESULTS: Radiologists thought that almost all performance measures were useful. They particularly liked seeing individual data presented in graphic form with a national benchmark or guideline for each performance measure clearly marked on the graph. They appreciated comparisons between their individual data and their peers' data (within their facility or state) and requested comparisons with national data (such as the BCSC). Many thought customizable, Web-based reports would be useful. CONCLUSION: Radiologists think that most audit statistics are useful; however, presenting performance data graphically with clear benchmarks may make them easier to understand. PMID- 19542409 TI - Improving patient safety: effects of a safety program on performance and culture in a department of radiology. AB - OBJECTIVE: Emphasis is being placed on improving the safety performance of the health care delivery system. The purpose of this study was to evaluate the effects of a program on safety performance and culture in a pediatric radiology department. MATERIALS AND METHODS: A comprehensive safety program implemented in a department of radiology included error prevention training for all employees, a safety coach program, safety awards, Crucial Conversations training, and operational rounds with radiology leaders. The number of serious safety events (events with deviation from best practice, patient harm, and causation) that in part involved radiology were compared for 2 years after implementation of the program and the previous 2 years (baseline). A U.S. Agency for Healthcare Research and Quality safety culture survey was distributed to radiology employees, and the responses were compared for periods early in the program and after full implementation of the program. Fisher's exact test was used to evaluate for statistically significant differences (p < 0.05) in the survey responses and the frequency of serious safety events. RESULTS: Before introduction of the safety program, radiology contributed to a serious safety event an average of once every 200 days as opposed to once in 780 days after implementation of the program (one event in more than two academic years) (p = 0.37). Improvement was found in all 12 dimensions of the culture survey after implementation of the program. Radiology scored higher than hospital averages in 10 of 12 dimensions of the survey. CONCLUSION: The safety program had a positive effect on safety culture. Although it is early in the process and proving statistical significance for rare events such as serious safety events is difficult, the mean number of days between serious safety events has increased from 200 to 780. We conclude that the program is having a positive effect on safety performance. PMID- 19542410 TI - Should you remove ionic, high-osmolality contrast media from your department? PMID- 19542411 TI - Anatomic study of the superficial peroneal nerve using sonography. AB - OBJECTIVE: The purpose of our study was to show that sonography allows precise assessment of the location and course of the superficial peroneal nerve and of its relationship with other structures. MATERIALS AND METHODS: This study, initially undertaken in cadavers, was followed by sonographic studies of 30 healthy adult volunteers (60 legs) by two radiologists in consensus. The location and course of the superficial peroneal nerve and its relationship with the adjacent anatomic structures were analyzed. RESULTS: The entire course of the superficial peroneal nerve could be identified using sonography. The level at which the superficial peroneal nerve emerges between the peroneus longus and extensor digitorum longus muscles and the level at which it pierces the crural fascia and becomes subcutaneous were found to be highly variable. The superficial peroneal nerve was found to be located in the anterior compartment in 26.7% of the legs and to divide before piercing the crural fascia in 6.7% of the legs. CONCLUSION: The superficial peroneal nerve can be clearly depicted by sonography. Knowledge of the nerve's precise location, which may show individual variations, may have useful clinical applications. PMID- 19542412 TI - Real-time sonoelastography of lateral epicondylitis: comparison of findings between patients and healthy volunteers. AB - OBJECTIVE: The purpose of this study was to evaluate real-time sonoelastography in the assessment of the origins the common extensor tendon in healthy volunteers and in patients with symptoms of lateral epicondylitis. The findings were compared with those obtained at clinical examination, ultrasonography, and power Doppler sonography. SUBJECTS AND METHODS: Thirty-eight elbows of 32 consecutively registered patients with symptoms of lateral epicondylitis and 44 asymptomatic elbows of 28 healthy volunteers were assessed with ultrasound and real-time sonoelastography. A clinical examination was performed, and pain was classified with a visual analog scale. RESULTS: In healthy volunteers, real-time sonoelastographic images showed hard tendon structures in 96% of tendon thirds and mild alterations in 4%. Real-time sonoelastography of patients showed hard structures in 33% of tendon thirds but softening of different grades in 67%, a statistically significant difference in relation to the findings in healthy volunteers (p < 0.001). Lateral collateral ligament involvement and overlying fascial involvement were more commonly detected with real-time sonoelastography. The sensitivity of real-time sonoelastography was 100%, the specificity 89%, and the accuracy 94% with clinical examination as the reference standard. Good correlation with ultrasound findings was found (r > or = 0.900). No correlation was observed between ultrasound or real-time sonoelastographic findings and power Doppler sonographic findings, but power Doppler sonographic findings had a strong correlation with the visual analog scale score. CONCLUSION: Real-time sonoelastography is valuable in the detection of the intratendinous and peritendinous alterations of lateral epicondylitis and facilitates differentiation between healthy and symptomatic extensor tendon origins with excellent sensitivity and excellent correlation with ultrasound findings. PMID- 19542413 TI - Ischiofemoral impingement syndrome: an entity with hip pain and abnormalities of the quadratus femoris muscle. AB - OBJECTIVE: The purpose of this study was to describe the MRI findings of an entity in which patients present with hip pain, abnormal MR signal intensity of the quadratus femoris muscle, and narrowing of the ischiofemoral space. MATERIALS AND METHODS: We reviewed MR images of 12 hips in nine patients with hip pain and abnormal MR signal intensity of the quadratus femoris muscle. Using axial MR images, two musculoskeletal radiologists measured the ischiofemoral and quadratus femoris spaces. We also examined changes to muscles and tendons for the presence of edema and tears. Data were compared with 11 hips in 10 control subjects. Statistical analyses determined interobserver variability and differences between groups. RESULTS: Subjects with an abnormal quadratus femoris muscle were all women 30-71 years old (mean age, 53 years) and had significantly narrower ischiofemoral spaces when compared with control subjects (13 +/- 5 vs 23 +/- 8 mm, respectively; p = 0.002). The quadratus femoris space was significantly narrower in affected subjects (7 +/- 3 vs 12 +/- 4 mm; p = 0.002). Abnormalities of the quadratus femoris muscle included edema (100%), partial tear (33%), and fatty infiltration (8%). The hamstring tendons of affected subjects showed evidence of edema (50%) and partial tears (25%). CONCLUSION: Ischiofemoral impingement may represent a cause of hip pain and should be considered in cases with MR signal abnormality of quadratus femoris muscle. PMID- 19542414 TI - Acute toxic leukoencephalopathy: potential for reversibility clinically and on MRI with diffusion-weighted and FLAIR imaging. AB - OBJECTIVE: Toxic leukoencephalopathy may present acutely or subacutely with symmetrically reduced diffusion in the periventricular and supraventricular white matter, hereafter referred to as periventricular white matter. This entity may reverse both on imaging and clinically. However, a gathering together of the heterogeneous causes of this disorder as seen on MRI with diffusion-weighted imaging (DWI) and an analysis of their likelihood to reverse has not yet been performed. Our goals were to gather causes of acute or subacute toxic leukoencephalopathy that can present with reduced diffusion of periventricular white matter in order to promote recognition of this entity, to evaluate whether DWI with apparent diffusion coefficient (ADC) values can predict the extent of chronic FLAIR abnormality (imaging reversibility), and to evaluate whether DWI can predict the clinical outcome (clinical reversibility). MATERIALS AND METHODS: Two neuroradiologists retrospectively reviewed the MRI examinations of 39 patients with acute symptoms and reduced diffusion of periventricular white matter. The reviewers then scored the extent of abnormality on DWI and FLAIR. ADC ratios of affected white matter versus the unaffected periventricular white matter were obtained. Each patient's clinical records were reviewed to determine the cause and clinical outcome. Histology findings were available in three patients. Correlations were calculated between the initial MRI markers and both the clinical course and the follow-up extent on FLAIR using Spearman's correlation coefficient. RESULTS: Of the initial 39 patients, seven were excluded because of a nontoxic cause (hypoxic-ischemic encephalopathy [HIE] or congenital genetic disorders) or because of technical errors. In the remaining 32 patients, no correlation was noted between any of the initial MRI markers (percentage of ADC reduction, DWI extent, or FLAIR extent) with the clinical outcome. Three patients had histologic correlation. However, moderate correlation was seen between the extent of abnormality on initial FLAIR and the extent on follow-up FLAIR (r = 0.441, p = 0.047). Of the 13 patients who underwent repeat MRI at 21 days or longer, the reduced diffusion resolved in all but one. Significant differences were noted between ADC values in affected white matter versus unaffected periventricular white matter on initial (p < 0.0001) but not on follow up MRI (p = 0.13), and in affected white matter on initial versus follow-up (p = 0.0014) in those individuals who underwent repeat imaging on the same magnet (n = 9), confirming resolution of the DWI abnormalities. CONCLUSION: Acute toxic leukoencephalopathy with reduced diffusion may be clinically reversible and radiologically reversible on DWI, and may also be reversible, but to a lesser degree, on FLAIR MRI. None of the imaging markers measured in this study appears to correlate with clinical outcome, which underscores the necessity for prompt recognition of this entity. Alerting the clinician to this potentially reversible syndrome can facilitate treatment and removal of the offending agent in the early stages. PMID- 19542416 TI - 18F-FDG PET evaluation of sinonasal papilloma. AB - OBJECTIVE: It has been suggested that (18)F-FDG uptake determined by PET can differentiate squamous cell carcinoma from benign sinonasal papilloma. We wish to present our experience with sinonasal papillomas and PET/CT to determine if the degree of FDG uptake is indicative of benign or malignant disease. CONCLUSION: Benign sinonasal papilloma may be associated with intense FDG uptake on PET/CT. FDG PET/CT does not appear to reliably differentiate benign from malignant sinonasal papilloma. PMID- 19542415 TI - Pattern recognition of benign nodules at ultrasound of the thyroid: which nodules can be left alone? AB - OBJECTIVE: The purpose of this study was to evaluate morphologic features predictive of benign thyroid nodules. MATERIALS AND METHODS: From a registry of the records of 1,232 fine-needle aspiration biopsies performed jointly by the cytology and radiology departments at a single institution between 2005 and 2007, the cases of 650 patients were identified for whom both a pathology report and ultrasound images were available. From the alphabetized list generated, the first 500 nodules were reviewed. We analyzed the accuracy of individual sonographic features and of 10 discrete recognizable morphologic patterns in the prediction of benign histologic findings. RESULTS: We found that grouping of thyroid nodules into reproducible patterns of morphology, or pattern recognition, rather than analysis of individual sonographic features, was extremely accurate in the identification of benign nodules. Four specific patterns were identified: spongiform configuration, cyst with colloid clot, giraffe pattern, and diffuse hyperechogenicity, which had a 100% specificity for benignity. In our series, identification of nodules with one of these four patterns could have obviated more than 60% of thyroid biopsies. CONCLUSION: Recognition of specific morphologic patterns is an accurate method of identifying benign thyroid nodules that do not require cytologic evaluation. Use of this approach may substantially decrease the number of unnecessary biopsy procedures. PMID- 19542417 TI - Vesselplasty: a new technical approach to treat symptomatic vertebral compression fractures. AB - OBJECTIVE: The objective of our study was to evaluate the effectiveness and safety of vesselplasty to treat symptomatic vertebral compression fractures (VCFs). SUBJECTS AND METHODS: Twenty-nine patients undergoing vesselplasty at our institution between April 2006 and February 2008 were enrolled in the study. All patients had been undergoing medical therapy for one or more painful VCFs. Pain, mobility, and analgesic use scores were obtained, and restoration of vertebral body height was evaluated. A two-tailed paired Student's t test was used to compare differences in the mean scores for levels of pain, mobility, and analgesic use before and after the procedure and to evaluate changes in vertebral body height. We analyzed the influence of the age of the fracture and its cause in the variations in the pain, mobility, and analgesic use scores. RESULTS: Seven of the 29 patients had fractures in more than one level, for a total of 37 procedures. The cause of the vertebral collapse was osteoporosis in 27 (73%), high-impact trauma in five (13.5%), myeloma in three (8%), and metastatic fracture in two (5.4%). The average pain score before treatment was 8.72 +/- 1.25 (SD), whereas the average pain score after treatment was 3.38 +/- 2.35. The average mobility score before treatment was 2.31 +/- 1.94, whereas the average mobility score after treatment was 0.59 +/- 1.05 (p < 0.001). The average analgesic use score before treatment was 3.07 +/- 1.46, whereas it was 1.86 +/- 1.90 after treatment (p < 0.001). There was no evidence of clinical complications. CONCLUSION: Vesselplasty offers statistically significant benefits in improvements of pain, mobility, and the need for analgesia in patients with symptomatic VCFs, thus providing a safe alternative in the treatment of these fractures. PMID- 19542418 TI - Survey of aorta and coronary arteries with prospective ECG-triggered 100-kV 64 MDCT angiography. AB - OBJECTIVE: The purpose of this study was to investigate the feasibility of prospective ECG-triggered axial 64-MDCT angiography of the aorta and coronary arteries performed at a tube voltage of 100 kV. SUBJECTS AND METHODS: Thirty patients with a heart rate less than 75 beats/min who were referred for aortic CT angiography were enrolled. The image quality of the ascending aorta, aortic valve, and coronary arteries was evaluated for motion artifacts. Contrast enhancement (mean attenuation) was measured in the ascending aorta, descending aorta, and bifurcation of the aorta. Aortic valve and coronary artery lesions were surveyed. RESULTS: Acceptable image quality was achieved in 100% (30/30) of cases for the ascending aorta, 97% (29/30) of cases for the aortic valve, and 98% (442/452) of coronary arterial segments. Contrast enhancement was greater than 200 HU and was satisfactory (ascending aorta, 379 +/- 80 HU; descending aorta, 354 +/- 72 HU; bifurcation, 355 +/- 96 HU). Lesions found in the aortic valve were plaque (n = 16) and bicuspid valve (n = 1) and in the coronary arteries were > or = 50% luminal stenosis (n = 5), plaque (n = 21), myocardial bridge (n = 12), and anomalous origin (n = 1). The effective radiation dose was estimated to be 7.5 +/- 1.7 mSv. CONCLUSION: For patients with a heart rate less than 75 beats/min, prospective ECG-triggered axial CT angiography at a tube voltage of 100 kV has the potential to provide clinically relevant information about the aorta and coronary arteries with low radiation exposure. PMID- 19542419 TI - Peripheral bolus-chase MR angiography: analysis of risk factors for nondiagnostic image quality of the calf vessels--a combined retrospective and prospective study. AB - OBJECTIVE: The objective of our study was to evaluate the influence of endogenous and exogenous risk factors on the rate of nondiagnostic examinations of the calves in peripheral bolus-chase MR angiography (MRA). SUBJECTS AND METHODS: Peripheral bolus-chase MRA runoff studies in 177 patients with peripheral arterial occlusive disease (PAOD) were retrospectively assessed with regard to the rate of nondiagnostic image quality due to substantial venous overlay in the calf arteries requiring repeated MRA examinations. Logistic regression was used to analyze the rate of nondiagnostic MRA examinations as a function of several endogenous and exogenous risk factors and of the stage of PAOD. To probe the retrospective data, 22 consecutive patients were prospectively included and underwent a standard peripheral MRA examination if the probability of a nondiagnostic examination was less than 50% based on the results of logistic regression; otherwise, a hybrid MRA examination was ordered. RESULTS: Nondiagnostic image quality of the calf arteries was found in 53 patients (30%). The incidence increased with each stage of PAOD up to 39% for stage IV. For each increase in the stage of PAOD, the probability of nondiagnostic image quality increased by a factor of 1.5561 (p = 0.0024). With an increasing number of risk factors, a significantly (p = 0.0074) higher rate of nondiagnostic images was found. CONCLUSION: Based on the retrospective statistical analysis of PAOD stages and risk factors, selected patients can be triaged to undergo a specific hybrid MRA technique and thus circumvent the occurrence of nondiagnostic images and the need for repeated MRA examinations. PMID- 19542420 TI - Metallic stent placement in the palliative treatment of malignant gastric outlet obstructions: primary gastric carcinoma versus pancreatic carcinoma. AB - OBJECTIVE: The objective of our study was to compare the clinical effectiveness of metallic stent placement for relief of gastric outlet obstruction caused by gastric carcinoma and pancreatic carcinoma. MATERIALS AND METHODS: A total of 207 patients with gastric outlet obstruction caused by inoperable gastric carcinoma (n = 147) or pancreatic carcinoma (n = 60) underwent metallic stent placement. RESULTS: Technical success of metallic stent placement was achieved in all patients. Clinical success was achieved in 97% and 93% of patients with gastric and pancreatic carcinoma, respectively (p = 0.286). The overall complication rate did not differ significantly between the gastric (29%) and pancreatic (23%) carcinoma groups (p = 0.441). Stent collapse was significantly more frequent in the gastric carcinoma group (11%) than the pancreatic carcinoma group (2%) (p = 0.027), whereas serious complications, including gastrointestinal bleeding and intestinal perforation, occurred more frequently in the pancreatic (7%) than the gastric (1%) carcinoma group (p = 0.026). The cumulative survival period was significantly longer in the gastric carcinoma (median, 153 days) than the pancreatic carcinoma (median, 90 days) group (p = 0.041), but cumulative stent patency did not differ significantly between the gastric carcinoma (median, 350 days) and pancreatic carcinoma (median, 385 days) groups (p = 0.415). CONCLUSION: Metallic stent placement was clinically effective in the palliative treatment of gastric outlet obstruction in patients with gastric and pancreatic carcinoma. The two groups differed significantly in the rates of stent collapse and serious complications and patient survival after stent placement. PMID- 19542421 TI - Dual-design expandable colorectal stent for malignant colorectal obstruction: comparison of flared ends and bent ends. AB - OBJECTIVE: The purpose of this study was to compare, focusing on colonic perforation and stent migration, the clinical safety and efficacy of dual-design expandable colorectal stents with flared ends with those of stents with bent ends in the treatment of patients with malignant colorectal obstruction. SUBJECTS AND METHODS: A total of 122 patients with malignant colorectal obstruction underwent implantation of dual-design stents with flared (n = 69) or bent (n = 53) ends. RESULTS: Stent placement was technically successful in 116 of 122 patients (95.1%), 65 of 69 patients (94.2%) with flared-end stents and 51 of 53 patients (96.2%) with bent-end stents (p > 0.05). Clinical success was achieved within 2 days in 61 of 65 patients (93.8%) with bent-end stents and in 46 of 51 patients (90.2%) with flared-end stents (p > 0.05). Complications included seven cases of colonic perforation (6%), seven cases of stent migration (6%), three cases of tumor overgrowth (2.6%), four cases of severe rectal pain (3.4%), and four cases of bleeding (3.4%). There were no significant differences between the rates of colonic perforation and stent migration in the two groups (6.2% vs 5.9%), and the overall complication rates were similar (p > 0.05). CONCLUSION: Dual-design expandable colorectal stents with flared ends and those with bent ends are equally safe and effective, having similar perforation and migration rates. PMID- 19542422 TI - Diffusion-weighted imaging of mucinous carcinoma of the breast: evaluation of apparent diffusion coefficient and signal intensity in correlation with histologic findings. AB - OBJECTIVE: The purposes of this study were to compare the apparent diffusion coefficient (ADC) of mucinous carcinoma of the breast with that of other breast tumors and to analyze correlations between signal intensity on diffusion-weighted images and the histologic features of mucinous carcinoma. SUBJECTS AND METHODS: Two hundred seventy-six patients with 277 lesions, including 15 mucinous carcinomas (13 pure type, two mixed type), 204 other malignant tumors, and 58 benign lesions, were examined with 1.5-T MRI at b values of 0 and 1,500 s/mm(2). The correlations between cellularity and ADC, homogeneity of signal intensity on diffusion-weighted images, and histopathologic findings were analyzed. The difference was statistically significant (p < 0.05). RESULTS: The mean ADC of mucinous carcinoma (1.8 +/- 0.4 x 10(-3) mm(2)/s) was statistically higher than that of benign lesions (1.3+/- 0.3 x 10(-3) mm(2)/s) and other malignant tumors (0.9 +/- 0.2 x 10(-3) mm(2)/s) (p < 0.001). The ADC of pure type mucinous carcinoma (1.8 +/- 0.3 x 10(-3) mm(2)/s) was higher than that of mixed type mucinous carcinoma (1.2 +/- 0.2 x 10(-3) mm(2)/s) (p < 0.001) and other histologic types (p > 0.05). The correlation between mean cellularity and the ADC of mucinous carcinoma was significant (rho(s) = -0.754; p = 0.001). The homogeneity of signal intensity on diffusion-weighted images correlated with the homogeneity of histologic structures of mucinous carcinoma (p < 0.001; kappa = 0.826). CONCLUSION: Mucinous carcinoma can be clearly differentiated from other breast tumors on the basis of ADC. The low signal intensity of mucinous carcinoma on diffusion-weighted images appears to reflect the presence of mucin and low cellularity. High signal intensity on diffusion-weighted images may reflect the presence of fibrovascular bundles, increased cell density, or a combination of these features. PMID- 19542424 TI - The original intrathymic progenitor from which T cells originate. PMID- 19542423 TI - Long-term quality of life assessment among patients undergoing uterine fibroid embolization. AB - OBJECTIVE: The purpose of this study was assessment of the long-term outcome of fibroid-associated quality of life among patients treated with uterine fibroid embolization. MATERIALS AND METHODS: A retrospective follow-up cohort study included all patients described in a 2006 publication. Analysis was performed with a questionnaire consisting of 49 questions about six topics. Assessment was focused on comparing symptoms and quality of life in long-term follow-up. RESULTS: The analysis was based on questionnaires completed by 39 patients. The median follow-up period was 7 years (interquartile range, 1.5 years). Uterine fibroid embolization led to a reduction of bleeding symptoms in 89.7% of the patients, pain in 78.9%, bulk-related symptoms in 89.5%, fatigue in 76.9%, limitations of social life in 92.9%, and depression in 78.6%. The median impairment scores for bleeding and pain decreased significantly from 7 to 0 and from 5 to 0 (both p < 0.001). The general quality-of-life index increased significantly from 4.5 to 9 (p < 0.001). In the long term, there was no significant difference in parameters assessed compared with the midterm follow-up findings. Six patients (15.4%) underwent hysterectomy an average of 32.1 months after intervention. Thirty-two patients (82.1%) continued to be satisfied with the intervention, and 30 patients (76.9%) answered that they would recommend uterine fibroid embolization to other patients. CONCLUSION: Uterine fibroid embolization seems to lead to notable long-term relief of fibroid-associated symptoms. In comparison with the midterm results, long-term outcome shows a clear continuance of improvement in general quality of life. PMID- 19542425 TI - Early T lymphocytes. Differentiation in vivo of adult intrathymic precursor cells. 1985. PMID- 19542427 TI - The effects of cytokines on suppression of lymphocyte proliferation by dexamethasone. AB - Treatment failure occurs in up to 30% of patients treated with steroids for inflammatory diseases. The aim of this study was to explore the potential role of 21 cytokines in steroid-resistant inflammatory disease and to develop methods to restore steroid sensitivity through cytokine manipulation. The dexamethasone inhibition of lymphocyte proliferation assay correlates with the outcome of steroid therapy in ulcerative colitis (UC) and other inflammatory diseases. Using this assay, PBMC production of 21 cytokines, assayed by cytokine bead array, was correlated with percentage of suppression of proliferation by 10(-6) M dexamethasone (Imax) in 26 healthy volunteers. Effects of the addition of exogenous cytokines to induce steroid resistance in PBMCs from healthy volunteers and cytokine blockade to improve steroid sensitivity in PBMCs from patients with steroid-resistant UC were then explored. Production of IL-1alpha, IL-10, IL-17, IFN-gamma, G-CSF, GM-CSF, TNF-alpha, and IFN-inducible protein 10 (IP-10) correlated significantly with in vitro steroid sensitivity; however, only IL-2 and TNF-alpha reduced steroid sensitivity when added exogenously. Addition of IL 10 enhanced steroid suppression. Immunoneutralization or receptor blockade of IL 2, but not TNF-alpha, IFN-gamma, IL-4, IL-17, or IP-10 increased steroid sensitivity in cells from steroid-resistant UC patients. Neutralization of IL-10 reduced steroid sensitivity. Of the large panel of cytokines studied, IL-2 appears to have the greatest antagonistic effect on the antiproliferative effect of steroids. These data suggest that IL-2 inhibition in vivo may improve the response to steroids in steroid-resistant individuals. PMID- 19542426 TI - Modulating the expression of IFN regulatory factor 8 alters the protumorigenic behavior of CD11b+Gr-1+ myeloid cells. AB - CD11b(+)Gr-1(+)-expressing cells, termed myeloid-derived suppressor cells, can mediate immunosuppression and tumor progression. However, the intrinsic molecular events that drive their protumorigenic behavior remain to be elucidated. Although CD11b(+)Gr-1(+) cells exist at low frequencies in normal mice, it also remains unresolved whether they are biologically distinct from those of tumor-bearing hosts. These objectives were investigated using CD11b(+)Gr-1(+) cells from both implantable (4T1) and autochthonous (mouse mammary tumor virus-polyomavirus middle T Ag (MMTV-PyMT)) mouse models of mammary carcinoma. Limited variation was observed in the expression of markers associated with immunoregulation between CD11b(+)Gr-1(+) cells of both tumor models, as well as with their respective controls (Cnt). Despite limited differences in phenotype, tumor-induced CD11b(+)Gr-1(+) cells were found to produce a more immunosuppressive cytokine profile than that observed by Cnt CD11b(+)Gr-1(+) cells. Furthermore, when admixed with tumor cells, CD11b(+)Gr-1(+) cells from tumor-bearing mice significantly enhanced neoplastic growth compared with counterpart cells from Cnt mice. However, the protumorigenic behavior of these tumor-induced CD11b(+)Gr-1(+) cells was significantly diminished when the expression of IFN regulatory factor 8, a key myeloid-associated transcription factor, was enhanced. The loss of this protumorigenic effect occurred independently of the host immune system and correlated with a CD11b(+)Gr-1(+) cytokine/chemokine production pattern that resembled cells from nontumor-bearing Cnt mice. Overall, our data indicate that 1) tumor-induced CD11b(+)Gr-1(+) cells from both cancer models were phenotypically similar, but biologically distinct from their nontumor-bearing counterparts and 2) modulation of IFN regulatory factor 8 levels in tumor-induced CD11b(+)Gr-1(+) cells can significantly abrogate their protumorigenic behavior, which may have important implications for cancer therapy. PMID- 19542428 TI - Role of CD44 in the differentiation of Th1 and Th2 cells: CD44-deficiency enhances the development of Th2 effectors in response to sheep RBC and chicken ovalbumin. AB - CD4 T cells can be primarily polarized to differentiate into Th1 or Th2 cells. CD44 is a marker of T cell activation and a property of long-lived memory cells and implicated in cell migration, activation, and differentiation. To date, whether CD44 has a role in regulating Th1-Th2 differentiation has not been determined. In this study, we compared Th1 and Th2 responses in wild-type and CD44-deficient mice in response to sheep RBC and chicken OVA, as well as examined Th1-Th2 differentiation in vivo and in vitro from CD44-sufficient and CD44 deficient naive CD4 T cells. We observed that deficiency of CD44 tended to inhibit Th1 while promoting Th2 differentiation. Furthermore, chimeric studies suggested that CD44 expression by CD4 T cells was essential for such Th2 bias. The regulation by CD44 occurred at the transcription level leading to up regulated GATA3 and down-regulated T-bet expression in activated CD4 T cells. We also noted that CD44-deficiency could modify the state of dendritic cell subsets to induce a Th2-biased development. Results presented in this study demonstrate for the first time that CD44 participates in the regulation of Th1-Th2 differentiation. PMID- 19542429 TI - Vaccine-induced CD8+ T cell-dependent suppression of airway hyperresponsiveness and inflammation. AB - Suppressing the abnormalities associated with asthma has been difficult to accomplish using immunotherapy or vaccination once the disease is established. The effector cells necessary for effective immunization/vaccination and immunotherapy of asthma are also not well understood. Therefore, we vaccinated allergen (OVA)-sensitized mice to determine whether therapeutic immunization could suppress airway hyperresponsiveness (AHR) and inflammation and to identify key immune effector cells and cytokines. Mice were immunized with a vaccine comprised of Ag and cationic liposome-DNA complexes (CLDC), a vaccine which has previously been shown to elicit strong CD4(+) and CD8(+) T cell responses and activation of Th1 immunity. We showed that immunization with the OVA-CLDC vaccine significantly suppressed AHR, eosinophilia, goblet cell metaplasia, and Th2 cytokine production. In contrast, immunization with CLDC alone suppressed eosinophilia and Th2 cytokine production, but failed to suppress AHR and goblet cell changes. Using adoptive transfer experiments, we found that suppression of AHR was mediated by Ag-specific CD8(+) T cells and was dependent on IFN-gamma production by the transferred T cells. Thus, we conclude that generation of strong, allergen-specific CD8(+) T cell responses by immunization may be capable of suppressing AHR and allergic airway inflammation, even in previously sensitized and challenged mice. PMID- 19542430 TI - A microbial polysaccharide reduces the severity of rheumatoid arthritis by influencing Th17 differentiation and proinflammatory cytokines production. AB - Rheumatoid arthritis (RA) is a chronic and debilitating autoimmune disease characterized by chronic joint inflammation with subsequent cartilage and bone destruction. RA is emerging as a model of IL-17-driven autoimmune inflammatory disease. IL-17 is a marker for Th17 cells, with its master regulator being the retinoic acid receptor-related orphan receptor (RORgammat) regulated by STAT3 signaling. Glucuronoxylomannan (GXM), a polysaccharide representing the main component of the capsular material of the opportunistic yeast Cryptococcus neoformans, exhibits potent immunosuppressive properties both in vitro and in vivo. The present study investigates the effects of GXM treatment on the progression of collagen-induced arthritis. GXM suppressed clinical signs of collagen-induced arthritis and blocked joint erosion progression. This effect was mediated by down-regulation of key cytokines involved in the pathogenesis of RA such as TNF-alpha and IL-1beta, and up-regulation of the inhibitory cytokine IL 10. Moreover, a reduction of IL-6 and TGF-beta, which inhibit Th17 differentiation with consequent decreased IL-17 production at the local and systemic level, was observed. The effect of GXM on Th17 differentiation mirrored the reduction in STAT3 activation and inhibition of RORgammat synthesis. Consequently, this work highlights the beneficial properties of an efficacious compound that could eventually be destined to the clinic. PMID- 19542431 TI - Cytokine-dependent modification of IL-12p70 and IL-23 balance in dendritic cells by ligand activation of Valpha24 invariant NKT cells. AB - CD1d-restricted invariant NKT (iNKT) cells play crucial roles in various types of immune responses, including autoimmune diseases, infectious diseases and tumor surveillance. The mechanisms underlying their adjuvant functions are well understood. Nevertheless, although IL-4 and IL-10 production characterize iNKT cells able to prevent or ameliorate some autoimmune diseases and inflammatory conditions, the precise mechanisms by which iNKT cells exert immune regulatory function remain elusive. This study demonstrates that the activation of human iNKT cells by their specific ligand alpha-galactosylceramide enhances IL-12p70 while inhibiting the IL-23 production by monocyte-derived dendritic cells, and in turn down-regulating the IL-17 production by memory CD4(+) Th cells. The ability of the iNKT cells to regulate the differential production of IL-12p70/IL-23 is mainly mediated by a remarkable hallmark of their function to produce both Th1 and Th2 cytokines. In particular, the down-regulation of IL-23 is markedly associated with a production of IL-4 and IL-10 from iNKT cells. Moreover, Th2 cytokines, such as IL-4 and IL-13 play a crucial role in defining the biased production of IL-12p70/IL-23 by enhancement of IL-12p70 in synergy with IFN gamma, whereas inhibition of the IFN-gamma-promoted IL-23 production. Collectively, the results suggest that iNKT cells modify the IL-12p70/IL-23 balance to enhance the IL-12p70-induced cell-mediated immunity and suppress the IL-23-dependent inflammatory pathologies. These results may account for the long appreciated contrasting beneficial and adverse consequence of ligand activation of iNKT cells. PMID- 19542432 TI - IFN-gamma attenuates antigen-induced overall immune response in the airway as a Th1-type immune regulatory cytokine. AB - Allergic inflammation in the airway is generally considered a Th2-type immune response. However, recent studies demonstrated that Th1- and Th17-type immune responses also play important roles in this process. IFN-gamma is a Th1-type cytokine that generally counteracts the Th2 response. Although previous studies suggest that exogenous IFN-gamma suppresses allergic airway inflammation, the mechanism of suppression has not been fully clarified. In this study, we elucidated whether IFN-gamma suppresses Ag-induced immune responses including the production of Th1- and Th17-type cytokines in the lung, and examined its mechanism of action. BALB/c mice were sensitized and challenged with OVA-Ag to induce airway inflammation. An IFN-gamma-producing plasmid vector was delivered before systemic Ag sensitization. IFN-gamma suppressed indicators of Th2-type immune responses such as airway eosinophilia, IL-5 and IL-13 production in the lung, and bronchial mucus production. Moreover, IFN-gamma also suppressed the production of IL-17 and IFN-gamma itself. The suppression was not mediated by inducing regulatory T cells or by inducing apoptosis in immunocytes. Instead, IFN gamma suppressed the Ag-presenting capacity and cytokine production of splenic dendritic cells and thus subsequently suppressed OVA-induced activation of CD4(+) T cells. Furthermore, IFN-gamma also attenuated allergic airway inflammation when delivered during the OVA challenge. Various functions of lung CD11c(+) APCs and their migration to regional lymph nodes were also suppressed. These results suggest that the Th1 cytokine IFN-gamma has broad immune regulatory potential through suppressing APC functions. They also suggest that delivery of IFN-gamma could be an effective strategy for regulating Ag-induced immune responses in the lung. PMID- 19542433 TI - Sphingosine kinase 1 is pivotal for Fc epsilon RI-mediated mast cell signaling and functional responses in vitro and in vivo. AB - Mast cell degranulation is pivotal to allergic diseases; investigating novel pathways triggering mast cell degranulation would undoubtedly have important therapeutic potential. FcepsilonRI-mediated degranulation has contradictorily been shown to require SphK1 or SphK2, depending on the reports. We investigated the in vitro and in vivo specific role(s) of SphK1 and SphK2 in FcepsilonRI mediated responses, using specific small interfering RNA-gene silencing. The small interfering RNA-knockdown of SphK1 in mast cells inhibited several signaling mechanisms and effector functions, triggered by FcepsilonRI stimulation including: Ca(2+) signals, NFkappaB activation, degranulation, cytokine/chemokine, and eicosanoid production, whereas silencing SphK2 had no effect at all. Moreover, silencing SPHK1 in vivo, in different strains of mice, strongly inhibited mast cell-mediated anaphylaxis, including inhibition of vascular permeability, tissue mast cell degranulation, changes in temperature, and serum histamine and cytokine levels, whereas silencing SPHK2 had no effect and the mice developed anaphylaxis. Our data differ from a recent report using SPHK1(-/-) and SPHK2(-/-) mice, which showed that SphK2 was required for FcepsilonRI-mediated mast cell responses. We performed experiments in mast cells derived from SPHK1(-/-) and SPHK2(-/-) mice and show that the calcium response and degranulation, triggered by FcepsilonRI-cross-linking, is not different from that triggered in wild-type cells. Moreover, IgE-mediated anaphylaxis in the knockout mice showed similar levels in temperature changes and serum histamine to that from wild-type mice, indicating that there was no protection from anaphylaxis for either knockout mice. Thus, our data strongly suggest a previously unrecognized compensatory mechanism in the knockout mice, and establishes a role for SphK1 in IgE-mediated mast cell responses. PMID- 19542434 TI - SHIP1 is a repressor of mast cell hyperplasia, cytokine production, and allergic inflammation in vivo. AB - SHIP1 inhibits immune receptor signaling through hydrolysis of the PI3K product phosphatidylinositol 3,4,5-trisphosphate, forming phosphatidylinositol 3,4 bisphosphate. In mast cells, SHIP1 represses FcepsilonRI- and cytokine-mediated activation in vitro, but little is known regarding the function of SHIP1 in mast cells in vivo or the susceptibility of Ship1(-/-) mice to mast cell-associated diseases. In this study, we found that Ship1(-/-) mice have systemic mast cell hyperplasia, increased serum levels of IL-6, TNF, and IL-5, and heightened anaphylactic response. Further, by reconstituting mast cell-deficient mice with Ship1(+/+) or Ship1(-/-) mast cells, we found that the above defects were due to loss of SHIP1 in mast cells. Additionally, we found that mice reconstituted with Ship1(-/-) mast cells suffered worse allergic asthma pathology than those reconstituted with Ship1(+/+) mast cells. In summary, our data show that SHIP1 represses allergic inflammation and mast cell hyperplasia in vivo and exerts these effects specifically in mast cells. PMID- 19542435 TI - Type V collagen-induced oral tolerance plus low-dose cyclosporine prevents rejection of MHC class I and II incompatible lung allografts. AB - Autoimmunity to type V collagen (col(V)) is a major risk factor for lung allograft rejection. Although col(V)-induced oral tolerance abrogates rejection of minor histoincompatible lung transplants, its ability to prevent rejection of fully MHC incompatible lung allografts is unknown. Rat lung allografts fully incompatible at MHC class I and II loci (Brown Norway (RT1(n))) were transplanted into untreated Wistar Kyoto rat recipients (WKY, RT1(l)), or WKY rats were fed col(V) pretransplantation. To determine whether col(V) enhanced cyclosporine (CsA)-mediated immune suppression, WKY rats were treated with low-dose CsA (5 mg/kg), posttransplant, or oral col(V) plus CsA. The data showed that in contrast to col(V) or CsA, col(V) plus low-dose CsA significantly prevented rejection pathology, down-regulated alloantigen-induced production of IFN-gamma and IL-17A, and suppressed chemotaxis for lung macrophages in allograft bronchoalveolar lavage fluid that was associated with lower local levels of MCP-1 (CCL2). Col(V) plus CsA was associated with alloantigen-induced expression of IL-10 in mediastinal lymph node or splenic T cells, intragraft expression of IL-10 and Foxp3 in perivascular and peribronchiolar mononuclear cells, and constitutive production of IL-10 from allograft alveolar macrophages. These data demonstrate that col(V) enhances low-dose CsA-mediated immune suppression, and suggest a role for oral col(V) in immune modulation in lung transplantation. PMID- 19542436 TI - Regulatory properties of copolymer I in Th17 differentiation by altering STAT3 phosphorylation. AB - Th17 and Th1 play an important role in multiple sclerosis for which copolymer I (COP-I) is a treatment option. We described here that the treatment effect of COP I correlated with its unique regulatory properties on differentiation and survival of Th17 in experimental autoimmune encephalomyelitis mice, which was mediated through down-regulation of STAT3 phosphorylation. The effect of COP-I on Th17 differentiation required CD14(+) monocytes through IL-6 signaling as a key mediator to regulate STAT3 phosphorylation and subsequent RORgammat expression in Th17 cells. The observed effect was markedly dampened when monocytes were genetically deficient for IL-6. Similar regulatory properties of COP-I were demonstrated in human Th17 differentiation. The study revealed the differential regulatory roles and the novel mechanism of action of COP-I chiefly responsible for its treatment efficacy in experimental autoimmune encephalomyelitis and multiple sclerosis. PMID- 19542437 TI - Production of both IL-27 and IFN-gamma after the treatment with a ligand for invariant NK T cells is responsible for the suppression of Th2 response and allergic inflammation in a mouse experimental asthma model. AB - Using an allergen-induced airway inflammation model, we show that an injection of alpha-galactosylceramide (alpha-GalCer), a ligand for invariant NK T (iNKT) cells, induced IL-27 and that this process is essential for the attenuation of the Th2 response. After the systemic administration of alpha-GalCer into the mice primed with OVA in alum, Th2 cytokine production of OVA-primed CD4(+) T cells in their lymph nodes, IgG1 and IgE Ab formation, and infiltration of eosinophils in bronchoalveolar lavage after the OVA challenge were suppressed. Systemic administration of rIFN-gamma into OVA-primed mice could not reproduce these effects of alpha-GalCer. IL-27p28 was detected both in the culture supernatant of alpha-GalCer-stimulated spleen cells and in the serum of the alpha-GalCer-treated mice, but not in the iNKT cell-deficient mice. Splenic iNKT cells produced IL 27p28 in the culture supernatant upon stimulation with PMA plus ionomycin, although the transcript of IL-27p28 in the iNKT cells was constitutively expressed regardless of the stimulation. By contrast, the transcript of IL-27EBI3 was induced in the iNKT cells upon stimulation with PMA plus ionomycin in vitro and with alpha-GalCer treatment in vivo, suggesting that IL-27 (p28/EBI3) could be produced by iNKT cells in an activation-dependent manner. Although repeated injections of rIL-27 did not substitute for the effects of a single injection of alpha-GalCer, administration of rIL-27 along with rIFN-gamma reproduced in vivo effects of the alpha-GalCer injection. These data indicate that production of both IL-27 and IFN-gamma by the alpha-GalCer treatment is responsible for suppression of the Th2 response and allergic inflammation. PMID- 19542439 TI - Gap junctions at the dendritic cell-T cell interface are key elements for antigen dependent T cell activation. AB - The acquired immune response begins with Ag presentation by dendritic cells (DCs) to naive T cells in a heterocellular cell-cell contact-dependent process. Although both DCs and T cells are known to express connexin43, a gap junction protein subunit, the role of connexin43 on the initiation of T cell responses remains to be elucidated. In the present work, we report the formation of gap junctions between DCs and T cells and their role on T cell activation during Ag presentation by DCs. In cocultures of DCs and T cells, Lucifer yellow microinjected into DCs is transferred to adjacent transgenic CD4(+) T cells, only if the specific antigenic peptide was present at least during the first 24 h of cocultures. This dye transfer was sensitive to gap junction blockers, such as oleamide, and small peptides containing the extracellular loop sequences of conexin. Furthermore, in this system, gap junction blockers drastically reduced T cell activation as reflected by lower proliferation, CD69 expression, and IL-2 secretion. This lower T cell activation produced by gap junction blockers was not due to a lower expression of CD80, CD86, CD40, and MHC-II on DCs. Furthermore, gap junction blocker did not affect polyclonal activation of T cell induced with anti-CD3 plus anti-CD28 Abs in the absence of DCs. These results strongly suggest that functional gap junctions assemble at the interface between DCs and T cells during Ag presentation and that they play an essential role in T cell activation. PMID- 19542438 TI - MEK/ERK-mediated phosphorylation of Bim is required to ensure survival of T and B lymphocytes during mitogenic stimulation. AB - Survival and death of lymphocytes are regulated by the balance between pro- and antiapoptotic members of the Bcl-2 family; this is coordinated with the control of cell cycling and differentiation. Bim, a proapoptotic BH3-only member of the Bcl-2 family, can be regulated by MEK/ERK-mediated phosphorylation, which affects its binding to pro-survival Bcl-2 family members and its turnover. We investigated Bim modifications in mouse B and T lymphoid cells after exposure to apoptotic stimuli and during mitogenic activation. Treatment with ionomycin or cytokine withdrawal caused an elevation in Bim(EL), the most abundant Bim isoform. In contrast, in mitogenically stimulated T and B cells, Bim(EL) was rapidly phosphorylated, and its levels declined. Pharmacological inhibitors of MEK/ERK signaling prevented both of these changes in Bim, reduced proliferation, and triggered apoptosis of mitogen-stimulated T and B cells. Loss of Bim prevented this cell killing but did not restore cell cycling. These results show that during mitogenic stimulation of T and B lymphocytes MEK/ERK signaling is critical for two distinct processes, cell survival, mediated (at least in part) through phosphorylation and consequent inhibition of Bim, and cell cycling, which proceeds independently of Bim inactivation. PMID- 19542440 TI - Altered thymic selection and increased autoimmunity caused by ectopic expression of DRAK2 during T cell development. AB - Negative regulation of TCR signaling is an important mechanism enforcing immunological self-tolerance to prevent inappropriate activation of T cells and thus the development of autoimmune diseases. The lymphoid-restricted serine/threonine kinase death-associated protein-related apoptotic kinase-2 (DRAK2) raises the TCR activation threshold by targeting TCR-induced calcium mobilization in thymocytes and peripheral T cells and regulates positive thymic selection and peripheral T cell activation. Despite a hypersensitivity of peripheral drak2-deficient T cells, drak2-deficient mice are enigmatically resistant to induced autoimmunity in the model experimental autoimmune encephalomyelitis. To further evaluate the differential role of DRAK2 in central vs peripheral tolerance and to assess its impact on the development of autoimmune diseases, we have generated a transgenic (Tg) mouse strain ectopically expressing DRAK2 via the lck proximal promoter (1017-DRAK2 Tg mice). This transgene led to highest expression levels in double-positive thymocytes that are normally devoid of DRAK2. 1017-DRAK2 Tg mice displayed a reduction of single-positive CD4(+) and CD8(+) thymocytes in context with diminished negative selection in male HY TCR x 1017-DRAK2 Tg mice as well as peripheral T cell hypersensitivity, enhanced susceptibility to experimental autoimmune encephalomyelitis, and spontaneous autoimmunity. These findings suggest that alteration in thymocyte signaling thresholds impacts the sensitivity of peripheral T cell pools. PMID- 19542441 TI - Early life exposure to lipopolysaccharide suppresses experimental autoimmune encephalomyelitis by promoting tolerogenic dendritic cells and regulatory T cells. AB - The rising incidence of autoimmune diseases such as multiple sclerosis (MS) in developed countries might be due to a more hygienic environment, particularly during early life. To investigate this concept, we developed a model of neonatal exposure to a common pathogen-associated molecular pattern, LPS, and determined its impact on experimental autoimmune encephalomyelitis (EAE). Mice exposed to LPS at 2 wk of age showed a delayed onset and diminished severity of myelin oligodendrocyte glycoprotein (MOG)-induced EAE, induced at 12 wk, compared with vehicle-exposed animals. Spinal cord transcript levels of CD3epsilon and F4/80 were lower in LPS- compared with PBS-exposed EAE animals with increased IL-10 levels in the LPS-exposed group. Splenic CD11c(+) cells from LPS-exposed animals exhibited reduced MHC class II and CD83 expression but increased levels of CD80 and CD86 both before and during EAE. MOG-treated APC from LPS-exposed animals stimulated less T lymphocyte proliferation but increased expansion of CD4(+)FoxP3(+) T cells compared with APC from PBS-exposed animals. Neuropathological studies disclosed reduced myelin and axonal loss in spinal cords from LPS-exposed compared with PBS-exposed animals with EAE, and this neuroprotective effect was associated with an increased number of CD3(+)FoxP3(+) immunoreactive cells. Analyses of human brain tissue revealed that FoxP3 expression was detected in lymphocytes, albeit reduced in MS compared with non-MS patients' brains. These findings support the concept of early-life microbial exposure influencing the generation of neuroprotective regulatory T cells and may provide insights into new immunotherapeutic strategies for MS. PMID- 19542442 TI - Vav1 regulates the migration and adhesion of dendritic cells. AB - Dendritic cells (DCs) are the most potent APCs for activating naive T cells, a process facilitated by the ability of immature DCs to mature and home to lymph nodes after encountering an inflammatory stimulus. Proteins involved in cytoskeletal rearrangement play an important role in regulating the adherence and motility of DCs. Vav1, a guanine nucleotide exchange factor for Rho family GTPases, mediates cytoskeletal rearrangement in hematopoietic cells following integrin ligation. We show that Vav1 is not required for the normal maturation of DCs in vitro; however, it is critical for DC binding to fibronectin and regulates the distribution but not the formation of podosomes. We also found that DC Vav1 was an important component of a signaling pathway involving focal adhesion kinase, phospholipase C-gamma2, and ERK1/2 following integrin ligation. Surprisingly, Vav1(-/-) DCs had increased rates of migration in vivo compared with wild-type control DCs. In vitro findings show that the presence of adhesive substrates such as fibronectin resulted in inhibition of migration. However, there was less inhibition in the absence of Vav1. These findings suggest that DC migration is negatively regulated by adhesion and integrin-mediated signaling and that Vav1 has a central role in this process. PMID- 19542443 TI - Clonotype selection and composition of human CD8 T cells specific for persistent herpes viruses varies with differentiation but is stable over time. AB - Protection from reactivation of persistent herpes virus infection is mediated by Ag-specific CD8 T cell responses, which are highly regulated by still poorly understood mechanisms. In this study, we analyzed differentiation and clonotypic dynamics of EBV- and CMV-specific T cells from healthy adults. Although these T lymphocytes included all subsets, from early-differentiated (EM/CD28(pos)) to late-differentiated (EMRA/CD28(neg)) stages, they varied in the sizes/proportions of these subsets. In-depth clonal composition analyses revealed TCR repertoires, which were highly restricted for CMV- and relatively diverse for EBV-specific cells. Virtually all virus-specific clonotypes identified in the EMRA/CD28(neg) subset were also found within the pool of less differentiated "memory" cells. However, striking differences in the patterns of dominance were observed among these subsets, because some clonotypes were selected with differentiation while others were not. Late-differentiated CMV-specific clonotypes were mostly characterized by TCR with lower dependency on CD8 coreceptor interaction. Yet all clonotypes displayed similar functional avidities, suggesting a compensatory role of CD8 in the clonotypes of lower TCR avidity. Importantly, clonotype selection and composition of each virus-specific subset upon differentiation was highly preserved over time, with the presence of the same dominant clonotypes at specific differentiation stages within a period of 4 years. Remarkably, clonotypic distribution was stable not only in late-differentiated but also in less-differentiated T cell subsets. Thus, T cell clonotypes segregate with differentiation, but the clonal composition once established is kept constant for at least several years. These findings reveal novel features of the highly sophisticated control of steady state protective T cell activity in healthy adults. PMID- 19542444 TI - The initial phase of an immune response functions to activate regulatory T cells. AB - An early reaction of CD4(+) T lymphocytes to Ag is the production of cytokines, notably IL-2. To detect cytokine-dependent responses, naive Ag-specific T cells were stimulated in vivo and the presence of phosphorylated STAT5 molecules was used to identify the cell populations responding to IL-2. Within hours of T cell priming, IL-2-dependent STAT5 phosphorylation occurred primarily in Foxp3(+) regulatory T cells. In contrast, the Ag-specific T cells received STAT5 signals only after repeated Ag exposure or memory differentiation. Regulatory T cells receiving IL-2 signals proliferated and developed enhanced suppressive activity. These results indicate that one of the earliest events in a T cell response is the activation of endogenous regulatory cells, potentially to prevent autoimmunity. PMID- 19542445 TI - Human placenta expresses and secretes NKG2D ligands via exosomes that down modulate the cognate receptor expression: evidence for immunosuppressive function. AB - During mammalian pregnancy maternal-fetal tolerance involves a number of immunosuppressive factors produced by placenta. Recently, placenta-derived exosomes have emerged as new immune regulators in the maternal immune tolerance. Exosomes are membrane nanovesicles with defined morphology, which are secreted from endosomal multivesicular bodies (MVB) upon fusion with the plasma membrane. Previously, we reported that the MHC class I chain-related (MIC) proteins A and B, human ligands of the activating NK cell receptor NKG2D, are expressed by placenta, sorted to MVB of syncytiotrophoblast and probably released via MIC bearing exosomes. In this report, we show that the second family of human NKG2D ligands, the UL-16 binding proteins (ULBP), is also expressed by placenta. Importantly, this expression was not due to placental CMV infection. Immunoelectron microscopy disclosed that ULBP1-5 are produced and retained in MVB of the syncytiotrophoblast on microvesicles/exosomes. Using human placenta explant cultures and different assays, we demonstrate that exosomes bearing NKG2D ligands are released by human placenta. Isolated placental exosomes carried ULBP1 5 and MIC on their surface and induced down-regulation of the NKG2D receptor on NK, CD8(+), and gammadelta T cells, leading to reduction of their in vitro cytotoxicity without affecting the perforin-mediated lytic pathway. Release of placental NKG2D ligands via exosomes is an alternative mechanism for generation of bioactive soluble form of these ligands. These findings highlight a role for NKG2D ligand-bearing placental exosomes in the fetal immune escape and support the view of placenta as a unique immunosuppressive organ. PMID- 19542446 TI - Notch ligands expressed by follicular dendritic cells protect germinal center B cells from apoptosis. AB - The Notch signaling pathway is one of the most conserved mechanisms to regulate cell fate in many tissues during development and postnatal life. In the immune system, Notch signaling regulates T and B cell development and modulates the differentiation of T and B cells. In this study, we investigated the functional roles of Notch signaling in human B cell differentiation within the germinal center (GC). Notch ligands, Delta-like 1 (Dll1) and Jagged 1 (Jg1), are expressed by follicular dendritic cells (FDC) but not by B cells in the GC, while GC-B cells express the Notch receptors, Notch1 and Notch2. The blockade of Notch signaling pathways using a gamma-secretase inhibitor, DAPT (N-[N-(3,5 difluorophenacetyl-l-alanyl)]-S-phenylglycine t-butyl ester), reduces the survival of GC-B cells in the presence of FDC/HK cells. Jg1 has a dominant effect on GC-B cell survival mediated by Notch signaling. Furthermore, Notch cooperates with another anti-apoptotic factor, BAFF/Blys produced by FDC to support GC-B cell growth. Taken together, our data shows the important role of Notch signaling provided by FDC in the survival of GC-B cells in vitro. PMID- 19542448 TI - Elimination of immunodominant epitopes from multispecific DNA-based vaccines allows induction of CD8 T cells that have a striking antiviral potential. AB - Immunodominance limits the TCR diversity of specific antiviral CD8 T cell responses elicited by vaccination or infection. To prime multispecific T cell responses, we constructed DNA vaccines that coexpress chimeric, multidomain Ags (with CD8 T cell-defined epitopes of the hepatitis B virus (HBV) surface (S), core (C), and polymerase (Pol) proteins and/or the OVA Ag as stress protein capturing fusion proteins. Priming of mono- or multispecific, HLA-A*0201- or K(b) restricted CD8 T cell responses by these DNA vaccines differed. K(b)/OVA(257-264) and K(b)/S(190-197)-specific CD8 T cell responses did not allow priming of a K(b)/C(93-100)-specific CD8 T cell response in mice immunized with multidomain vaccines. Tolerance to the S- Ag in transgenic Alb/HBs mice (that express large amounts of transgene-encoded S- Ag in the liver) facilitated priming of subdominant, K(b)/C(93-100)-specific CD8 T cell immunity by multidomain Ags. The "weak" (i.e., easily suppressed) K(b)/C(93-100)-specific CD8 T cell response was efficiently elicited by a HBV core Ag-encoding vector in 1.4HBV-S(mut) tg mice (that harbor a replicating HBV genome that produces HBV surface, core, and precore Ag in the liver). K(b)/C(93-100)-specific CD8 T cells accumulated in the liver of vaccinated 1.4HBV-S(mut) transgenic mice where they suppressed HBV replication. Subdominant epitopes in vaccines can hence prime specific CD8 T cell immunity in a tolerogenic milieu that delivers specific antiviral effects to HBV expressing hepatocytes. PMID- 19542447 TI - Activated integrin VLA-4 localizes to the lamellipodia and mediates T cell migration on VCAM-1. AB - Lymphocyte migration from blood into lymphoid tissues or to sites of inflammation occurs through interactions between cell surface integrins and their ligands expressed on the vascular endothelium and the extracellular matrix. VLA-4 (alpha(4)beta(1)) is a key integrin in the effective trafficking of lymphocytes. Although it has been well established that integrins undergo functionally significant conformational changes to mediate cell adhesion, there is no mechanistic information that explains how these are dynamically and spatially regulated during lymphocyte polarization and migration. Using dynamic fluorescence resonance energy transfer analysis of a novel VLA-4 FRET sensor under total internal reflection fluorescence microscopy, we show that VLA-4 activation localizes to the lamellipodium in living cells. During T cell migration on VCAM-1, VLA-4 activation concurs with spatial redistribution of chemokine receptor and active Rap1 at the leading edge. Selective inhibition of the activated VLA-4 at the leading edge with a small molecule inhibitor is sufficient to block T cell migration. These data suggest that a subpopulation of activated VLA-4 is mainly localized to the leading edge of polarized human T cells and is critical for T cell migration on VCAM-1. PMID- 19542449 TI - IL-23 promotes the production of IL-17 by antigen-specific CD8 T cells in the absence of IL-12 and type-I interferons. AB - In contrast to CD4 T cells, CD8 T cells inherently differentiate into IFN-gamma producing effectors. Accordingly, while generation of IFN-gamma-producing Th1 CD4 T cells was profoundly impaired in mice deficient for both type-I IFN and IL-12 signaling in response to infection with Listeria monocytogenes, generation of Ag specific, IFN-gamma-producing CD8 T cells was unimpaired. However, a fraction of these CD8 T cells also produced IL-17 in an IL-23-dependent manner. Furthermore, the addition of IL-23 in vitro was sufficient for some naive CD8 T cells to differentiate into IFN-gamma/IL-17 dual-producing cells and was associated with increased expression of ROR-gammat and ROR-alpha. Addition of IL-6 and TGF-beta to IL-23 further augmented ROR-gammat and ROR-alpha expression and suppressed Eomes expression, thereby enhancing IL-17 production by CD8 T cells. A loss of cytotoxic function accompanied the production of IL-17, as the addition of IL-6 and TGF-beta resulted in a marked reduction of granzyme B and perforin expression. Thus, CD8 T cells retain sufficient plasticity to respond to environmental cues and can acquire additional effector functions in response to their environmental context. PMID- 19542450 TI - Increased intracellular, cell surface, and secreted inducible heat shock protein 70 responses are triggered during the monocyte to dendritic cell (DC) transition by cytokines independently of heat stress and infection and may positively regulate DC growth. AB - Physiologic triggers and functional consequences of endogenous heat shock protein (HSP) responses in dendritic cells (DC) are poorly defined. In this study, we show that even in the absence of heat stress and infection, a specific cohort of DC/proinflammatory cytokines (IL-4-IL-13/IL-6/GM-CSF) institutes an enhanced inducible (i)HSP70 intracellular and extracellular response in human monocyte derived DC, especially during the monocyte to DC transition. Interestingly, whereas heat stress alone initiated an intracellular iHSP70 response in monocyte DC precursors, it did not promote cell surface or secreted iHSP70 responses, both of which were induced by cytokines independently of heat. The cytokine-induced iHSP70 response, which did not occur in lymphocytes, or monocytes-macrophages generated with M-CSF, was instituted within 48 h of cytokine exposure, and peaked upon commitment to DC growth at 72 h. Although a return to baseline levels was noted after this period, a distinct rise in iHSP70 occurred again during terminal DC maturation. Chemical inhibition of the iHSP70 response with either triptolide or KNK-437 was coupled with inhibition of DC differentiation and yielded cells displaying features of monocytes-macrophages. Exogenously supplied riHSP70 amplified events associated with cytokine-advanced DC differentiation/maturation, most notably the up-regulation of antiapoptotic proteins (Bcl-x(L)). Engaging the HSP receptor CD40 with CD40L produced identical results as extracellular riHSP70, and, moreover, an enhanced iHSP70 response. Thus, distinct iHSP70 and HSP receptor-mediated responses are triggered by cytokines irrespective of heat stress and infection in monocyte-derived DC and may function to positively regulate monocyte-derived DC, especially during critical periods of their growth. PMID- 19542451 TI - CD11c+CD8alpha+ dendritic cells promote protective immunity to respiratory infection with Bordetella pertussis. AB - CD11c(+)CD8alpha(+) and CD103(+) dendritic cells (DC) have been shown to promote regulatory T cell responses and mediate tolerance in the gastrointestinal tract. These cells have also been identified in the lung, but their role in immunity to respiratory tract infection is not clear. In this study, we have used a murine model of infection with Bordetella pertussis to examine the function of DC subtypes in protective immunity in the lungs. We found a dramatic increase in the numbers of CD11c(+)CD8alpha(+) DC in the cervical lymph nodes within 4 h of challenge with B. pertussis and these DC could acquire particulate Ag from the upper respiratory tract. CD11c(+)CD8alpha(+) DC also infiltrated the lung with a peak 7 days after B. pertussis challenge. The infiltrating CD11c(+)CD8alpha(+) DC expressed MHC, costimulatory and activation markers indicative of mature DC. The CD11c(+)CD8alpha(+) DC in the cervical lymph nodes expressed IL-4 and IL-10 and lower levels of IFN-gamma, but in the lungs expressed predominantly IFN-gamma. Depletion of CD8alpha(+) cells early in infection attenuated Th1 responses in the lungs and significantly reduced bacterial clearance. Conversely, transfer of FLT3 ligand (FL)-expanded CD11c(+)CD8alpha(+) DC enhanced bacterial clearance, whereas GM-CSF-expanded conventional DC had no effect. The numbers of CD11c(+)CD8alpha(+)CD103(+) cells were also increased during the early phase of infection. Blocking CD103 function caused a significant delay in bacterial clearance and a reduction in cellular infiltration into the lungs. These findings demonstrate that CD11c(+)CD8alpha(+) and CD11c(+)CD103(+)DC play a protective role in mediating immunity to B. pertussis infection in the respiratory tract. PMID- 19542452 TI - Neutrophil elastase represses IL-8/CXCL8 synthesis in human airway smooth muscle cells through induction of NF-kappa B repressing factor. AB - NF-kappaB repressing factor (NRF), a nuclear inhibitor of NF-kappaB, is constitutively expressed and is implicated in the basal silencing of specific NF kappaB targeting genes, including IFN-beta, IL-8/CXCL8, and iNOS. Little is known about the regulation of NRF and its role in response to stimuli. Airway smooth muscle (ASM) is a rich source of inflammatory mediators that may regulate the development and progression of airway inflammation. We have previously reported that NE activates NF-kappaB in primary human ASM (hASM), leading to induction of TGF-beta1. In this study, we describe that, instead of inducing the NF-kappaB response gene IL-8/CXCL8, NE suppressed IL-8/CXCL8 release and mRNA expression in hASM cells. Transcriptional blockade studies using actinomycin D revealed a similar degradation rate of IL-8/CXCL8 mRNA in the presence or absence of NE, suggesting an involvement at the transcription level. Mechanistically, the NE repressive effect was mediated by inducing NRF, as shown by RT-PCR and Western blotting, which was subsequently recruited to the native IL-8/CXCL8 promoter leading to removal of RNA polymerase II from the promoter, as demonstrated by chromatin immunoprecipitation assays. Knockdown of NRF by small interfering RNA prevented NE-induced suppression of IL-8/CXCL8 expression. In contrast, NE did not induce NRF expression in A549 and Beas-2B cells, where NE only stimulates NF kappaB activation and IL-8/CXCL8 induction. Forced expression of NRF in A549 cells by an NRF expression plasmid suppressed IL-8/CXCL8 expression. Hence, we describe a novel negative regulatory mechanism of NE-induced NRF, which is restricted to hASM and mediates the suppression of IL-8/CXCL8 expression. PMID- 19542453 TI - Structural basis for proteolytic specificity of the human apoptosis-inducing granzyme M. AB - Granzyme M (GzmM), a unique serine protease constitutively expressed in NK cells, is important for granule-mediated cytolysis and can induce rapid caspase dependent apoptosis of tumor cells. However, few substrates of GzmM have been reported to date, and the mechanism by which this enzyme recognizes and hydrolyzes substrates is unknown. To provide structural insights into the proteolytic specificity of human GzmM (hGzmM), crystal structures of wild-type hGzmM, the inactive D86N-GzmM mutant with bound peptide substrate, and the complexes with a catalytic product and with a tetrapeptide chloromethylketone inhibitor were solved to 1.96 A, 2.30 A, 2.17 A and 2.70 A, respectively. Structure-based mutagenesis revealed that the N terminus and catalytic triad of hGzmM are most essential for proteolytic function. In particular, D86N-GzmM was found to be an ideal inactive enzyme for functional studies. Structural comparisons indicated a large conformational change of the L3 loop upon substrate binding, and suggest this loop mediates the substrate specificity of hGzmM. Based on the complex structure of GzmM with its catalytic product, a tetrapeptide chloromethylketone inhibitor was designed and found to specifically block the catalytic activity of hGzmM. PMID- 19542455 TI - The transmembrane E3 ligase GRAIL ubiquitinates and degrades CD83 on CD4 T cells. AB - Ubiquitination of eukaryotic proteins regulates a broad range of cellular processes, including T cell activation and tolerance. We have previously demonstrated that GRAIL (gene related to anergy in lymphocytes), a transmembrane RING finger ubiquitin E3 ligase, initially described as induced during the induction of CD4 T cell anergy, is also expressed in resting CD4 T cells. In this study, we show that GRAIL can down-modulate the expression of CD83 (previously described as a cell surface marker for mature dendritic cells) on CD4 T cells. GRAIL-mediated down-modulation of CD83 is dependent on an intact GRAIL extracellular protease-associated domain and an enzymatically active cytosolic RING domain, and proceeds via the ubiquitin-dependent 26S proteosome pathway. Ubiquitin modification of lysine residues K168 and K183, but not K192, in the cytoplasmic domain of CD83 was shown to be necessary for GRAIL-mediated degradation of CD83. Reduced CD83 surface expression levels were seen both on anergized CD4 T cells and following GRAIL expression by retroviral transduction, whereas GRAIL knock-down by RNA interference in CD4 T cells resulted in elevated CD83 levels. Furthermore, CD83 expression on CD4 T cells contributes to T cell activation as a costimulatory molecule. This study supports the novel mechanism of ubiquitination by GRAIL, identifies CD83 as a substrate of GRAIL, and ascribes a role for CD83 in CD4 T cell activation. PMID- 19542454 TI - Structural bases for the affinity-driven selection of a public TCR against a dominant human cytomegalovirus epitope. AB - Protective T cell responses elicited along chronic human CMV (HCMV) infections are sometimes dominated by CD8 T cell clones bearing highly related or identical public TCR in unrelated individuals. To understand the principles that guide emergence of these public T cell responses, we have performed structural, biophysical, and functional analyses of an immunodominant public TCR (RA14) directed against a major HLA-A*0201-restricted HCMV Ag (pp65(495-503)) and selected in vivo from a diverse repertoire after chronic stimulations. Unlike the two immunodominant public TCRs crystallized so far, which focused on one peptide hotspot, the HCMV-specific RA14 TCR interacts with the full array of available peptide residues. The conservation of some peptide-MHC complex-contacting amino acids by lower-affinity TCRs suggests a shared TCR-peptide-MHC complex docking mode and supports an Ag-driven selection of optimal TCRs. Therefore, the emergence of a public TCR of an oligoclonal Ag-specific response after repeated viral stimulations is based on a receptor displaying a high structural complementarity with the entire peptide and focusing on three peptide hotspots. This highlights key parameters underlying the selection of a protective T cell response against HCMV infection, which remains a major health issue in patients undergoing bone marrow transplantation. PMID- 19542456 TI - The coding ECP 434(G>C) gene polymorphism determines the cytotoxicity of ECP but has minor effects on fibroblast-mediated gel contraction and no effect on RNase activity. AB - Eosinophil cationic protein (ECP) is a secretory protein of the eosinophil granulocyte, a cell involved in innate immunity. Functional studies have implicated ECP in numerous processes, such as tissue remodeling in allergic inflammation and cytotoxicity toward a variety of pathogens. Recent genetic studies have suggested that the ECP 434(G>C) polymorphism resulting in an arg97thr substitution would alter the function of ECP in vivo. Functional (in vitro) studies of ECP up until now have either been conducted with native preparations containing an unknown mixture of the ECP(97arg) and ECP(97thr) variants, or with recombinant proteins. Therefore, we have now for the first time extracted the native ECP(97arg) and ECP(97thr) variants from healthy blood donors and tested them functionally in vitro. Our results show that the arg97thr shift dramatically alters the cytotoxic capacity of ECP in vitro; the tested ECP(97arg) variants were cytotoxic toward the small-cell lung cancer cell line NCI-H69, whereas ECP(97thr) was noncytotoxic. RNase activity was unaffected by the arg97thr substitution. Both ECP(97arg) and ECP(97thr) stimulated fibroblast mediated collagen gel contraction, an experimental model, which depicts wound healing, in a dose-dependent manner. In conclusion, our results demonstrate that the ECP 434(G>C) gene polymorphism affects the functional properties of native ECP, but also that there is a dissociation between different biological activities; the arg97thr substitution impairs the cytotoxic potential of ECP but less the gel contraction and not at all the RNase activity. PMID- 19542457 TI - Antibodies in a heavy chain knock-in mouse exhibit characteristics of early heavy chain rearrangement. AB - Studies in autoantibody transgenic mice have demonstrated receptor editing rearrangements at Ab H and L chain loci. However, the physiologic role of H chain editing (V(H) replacement and rearrangement on the second allele) has been called into question. It is unclear if additional rounds of H chain rearrangement are driven by BCR specificity. In this study, we analyze the manner in which B cells undergo additional H chain rearrangements in an anti-DNA H chain knock-in mouse, B6.56R. We find that rearrangements in 56R(+) B cells tend to involve the D gene locus on both alleles and the most J(H)-proximal V(H) gene segments on the endogenous allele. As a result, some B cells exhibit V(D)J rearrangements on both H chain alleles, yet allelic exclusion is tightly maintained in mature 56R B cells. As B cells mature, a higher proportion expresses the nontransgenic H chain allele. Rearrangements on both H chain alleles exhibit junctional diversity consistent with TdT-mediated N-addition, and TdT RNA is expressed exclusively at the pro-B cell stage in B6.56R. Collectively, these findings favor a single, early window of H chain rearrangement in B6.56R that precedes the expression of a functional BCR. B cells that happen to successfully rearrange another H chain may be favored in the periphery. PMID- 19542458 TI - Immunization with the DNA-encoding N-terminal domain of proteophosphoglycan of Leishmania donovani generates Th1-type immunoprotective response against experimental visceral leishmaniasis. AB - Leishmania produce several types of mucin-like glycoproteins called proteophosphoglycans (PPGs) which exist as secretory as well as surface-bound forms in both promastigotes and amastigotes. The structure and function of PPGs have been reported to be species and stage specific as in the case of Leishmania major and Leishmania mexicana; there has been no such information available for Leishmania donovani. We have recently demonstrated that PPG is differentially expressed in sodium stibogluconate-sensitive and -resistant clinical isolates of L. donovani. To further elucidate the structure and function of the ppg gene of L. donovani, a partial sequence of its N-terminal domain of 1.6 kb containing the majority of antigenic determinants, was successfully cloned and expressed in prokaryotic as well as mammalian cells. We further evaluated the DNA-encoding N terminal domain of the ppg gene as a vaccine in golden hamsters (Mesocricetus auratus) against the L. donovani challenge. The prophylactic efficacy to the tune of approximately 80% was observed in vaccinated hamsters and all of them could survive beyond 6 mo after challenge. The efficacy was supported by a surge in inducible NO synthase, IFN-gamma, TNF-alpha, and IL-12 mRNA levels along with extreme down-regulation of TGF-beta, IL-4, and IL-10. A rise in the level of Leishmania-specific IgG2 was also observed which was indicative of enhanced cellular immune response. The results suggest the N-terminal domain of L. donovani ppg as a potential DNA vaccine against visceral leishmaniasis. PMID- 19542459 TI - Multivalent binding of carbohydrates by the human alpha-defensin, HD5. AB - Four of the six human alpha-defensins (human neutrophil peptides 1-3 and human alpha-defensin 5; HD5) have a lectin-like ability to bind glycosylated proteins. Using HD5 as a model, we applied surface plasmon resonance techniques to gain insights into this property. HD5 bound natural glycoproteins > neoglycoproteins based on BSA > nonglycosylated BSA >> free sugars. The affinity of HD5 for simple sugars covalently bound to BSA was orders of magnitude greater than its affinity for the same sugars in solution. The affinity of HD5 for protein-bound carbohydrates resulted from multivalent interactions which may also involve noncarbohydrate residues of the proteins. HD5 showed concentration-dependent self association that began at submicromolar concentrations and proceeded to dimer and tetramer formation at concentrations below 5 microM. The (R9A, R28A) and (R13A, R32A) analogs of HD5 showed greatly reduced self-association as well as minimal binding to BSA and to BSA-affixed sugars. From this and other evidence, we conclude that the extensive binding of HD5 to (neo)glycoproteins results from multivalent nonspecific interactions of individual HD5 molecules with carbohydrate and noncarbohydrate moieties of the target molecule and that the primary binding events are magnified and enhanced by subsequent in situ assembly and oligomerization of HD5. Self-association and multivalent binding may play integral roles in the ability of HD5 to protect against infections caused by viruses and other infectious agents. PMID- 19542460 TI - Downstream signals for MyD88-mediated phagocytosis of Borrelia burgdorferi can be initiated by TRIF and are dependent on PI3K. AB - We previously have shown that MyD88 is important for uptake of Borrelia burgdorferi by bone marrow derived macrophages (BMDMs). The mechanism by which MyD88 is involved in uptake of B. burgdorferi is currently is not well characterized. Here, we report that MyD88-mediated defect in the phagocytosis of B. burgdorferi can be complemented by TLR3/Toll/IL-1R domain-containing adaptor inducing IFN-beta (TRIF) activation in BMDMs from MyD88(-/-) mice. This effect of TLR3/TRIF activation was not due to its induction of type I IFNs, suggesting instead a convergence of signaling pathways downstream of MyD88 and TRIF. To characterize signaling pathways involved in MyD88-mediated phagocytosis of B. burgdorferi, BMDMs were treated with specific inhibitors of MAPK, protein kinase C, JAK/STAT, or PI3K. Only inhibition of PI3K resulted in a significant decrease of B. burgdorferi uptake. Consistent with this, B. burgdorferi activation of MyD88 or TLR3/TRIF signaling resulted in increased activity of PI3K. Additionally, association of B. burgdorferi with actin-related protein (Arp2/3) complexes, which facilitate actin rearrangements during phagocytosis, was similarly reduced in MyD88(-/-) BMDMs and in BMDMs treated with a PI3K inhibitor. Taken together, these findings define an essential pathway whereby downstream signals from MyD88 or TRIF converge on PI3K, which triggers actin polymerization to initiate the phagocytosis of B. burgdorferi. PMID- 19542461 TI - Soluble TLR2 reduces inflammation without compromising bacterial clearance by disrupting TLR2 triggering. AB - TLR overactivation may lead to end organ damage and serious acute and chronic inflammatory conditions. TLR responses must therefore be tightly regulated to control disease outcomes. We show in this study the ability of the soluble form of TLR2 (sTLR2) to regulate proinflammatory responses, and demonstrate the mechanisms underlying sTLR2 regulatory capacity. Cells overexpressing sTLR2, or stimulated in the presence of the sTLR2 protein, are hyporesponsive to TLR2 ligands. Regulation was TLR2 specific, and affected NF-kappaB activation, phagocytosis, and superoxide production. Natural sTLR2-depleted serum rendered leukocytes hypersensitive to TLR2-mediated stimulation. Mice administered sTLR2 together with Gram-positive bacteria-derived components showed lower peritoneal levels of the neutrophil (PMN) chemoattractant, keratinocyte-derived chemokine; lower PMN numbers; and a reduction in late apoptotic PMN. Mononuclear cell recruitment remained unaffected, and endogenous peritoneal sTLR2 levels increased. Notably, the capacity of sTLR2 to modulate acute inflammatory parameters did not compromise the ability of mice to clear live Gram-positive bacteria-induced infection. Mechanistically, sTLR2 interfered with TLR2 mobilization to lipid rafts for signaling, acted as a decoy microbial receptor, and disrupted the interaction of TLR2 with its coreceptor, CD14, by associating with CD14. These findings establish sTLR2 as a regulator of TLR2-mediated inflammatory responses, capable of blunting immune responses without abrogating microbial recognition and may inform the design of novel therapeutics against acute and chronic inflammatory conditions. PMID- 19542462 TI - Generation of protective T cell-independent antiviral antibody responses in SCID mice reconstituted with follicular or marginal zone B cells. AB - B cells generated in the bone marrow of adult mice enter the periphery as transitional B cells and subsequently differentiate into one of two phenotypically and functionally distinct subsets, marginal zone (MZ) or follicular (Fo) B cells. Recent reports indicate, however, that in response to environmental cues, such as lymphopenia, mature Fo B cells can change to display phenotypic markers characteristic of MZ B cells. Previously, we found that splenic B cells transferred to SCID mice responded to polyoma virus (PyV) infection with T cell-independent (TI) IgM and IgG secretion, reducing the viral load and protecting mice from the lethal effect of the infection. The contribution of MZ and Fo B cell subsets to this antiviral TI-2 response, however, has not been addressed. In this study, we show that both sort-purified MZ and Fo B cells generate protective TI Ab responses to PyV infection when transferred into SCID mice. Moreover, the transferred Fo B cells in the spleens of the PyV-infected SCID mice change phenotype, with many of them displaying MZ B cell characteristics. These findings demonstrate the plasticity of the B cell subsets in virus-infected hosts and show for the first time that B cells derived exclusively from Fo B cells can effectively function in antiviral TI-2 responses. PMID- 19542463 TI - Follicular dendritic cells activate HIV-1 replication in monocytes/macrophages through a juxtacrine mechanism mediated by P-selectin glycoprotein ligand 1. AB - Follicular dendritic cells (FDCs) are located in the lymphoid follicles of secondary lymphoid tissues and play a pivotal role in the selection of memory B lymphocytes within the germinal center, a major site for HIV-1 infection. Germinal centers are composed of highly activated B cells, macrophages, CD4(+)T cells, and FDCs. However, the physiological role of FDCs in HIV-1 replication remains largely unknown. We demonstrate in our current study that FDCs can efficiently activate HIV-1 replication in latently infected monocytic cells via an intercellular communication network mediated by the P-selectin/P-selectin glycoprotein ligand 1 (PSGL-1) interaction. Upon coculture with FDCs, HIV-1 replication was significantly induced in infected monocytic cell lines, primary monocytes, or macrophages. These cocultures were found to synergistically induce the expression of P-selectin in FDCs via NF-kappaB activation and its cognate receptor PSGL-1 in HIV-1-infected cells. Consistent with this observation, we find that this response is significantly blocked by antagonistic Abs against PSGL 1 and almost completely inhibited by PSGL-1 small interfering RNA. Moreover, a selective inhibitor for Syk, which is a downstream effector of PSGL-1, blocked HIV-1 replication in our cultures. We have thus elucidated a novel regulatory mechanism in which FDCs are a potent positive bystander that facilitates HIV-1 replication in adjacent infected monocytic cells via a juxtacrine signaling mechanism. PMID- 19542464 TI - In vivo lipopolysaccharide exposure of human blood leukocytes induces cross tolerance to multiple TLR ligands. AB - In vitro and in vivo experiments in mice have shown that exposure of cells to the TLR4 ligand LPS induces tolerance toward a second exposure to LPS and induces cross-tolerance to certain other TLR ligands. Recently, we found that LPS tolerance in experimental human endotoxemia and Gram-negative sepsis is associated with elevated levels of IL-1R-associated kinase M, an intracellular negative regulator of MyD88-dependent TLR signaling. In the present study, we investigated whether in vivo exposure of humans to LPS induces tolerance in circulating leukocytes to other TLR agonists that rely either on MyD88- dependent or on MyD88-independent signaling. Analysis of TNF, IL-1beta, IL-6, and IL-10 levels in whole blood demonstrated that leukocytes were hyporesponsive to ex vivo LPS restimulation 3-8 h after i.v. LPS injection (4 ng/kg). Reduced cytokine release during the same interval was also observed in whole blood further stimulated with MyD88-dependent ligands for TLR2, TLR5, and TLR7 or with whole bacteria. Strikingly, blood leukocytes were also tolerant to a ligand for TLR3, which signals solely through a MyD88-independent (Toll IL-1R domain-containing adaptor-inducing IFN-beta (TRIF)-dependent) pathway. The hyporesponsiveness of leukocytes to TLR3 ligation was associated with reduced rather than increased levels of the recently identified TRIF inhibitor SARM. Taken together, these data indicate that systemic LPS challenge of human volunteers induces cross-tolerance to multiple TLR ligands that signal in a MyD88-dependent or MyD88-independent manner and suggest that LPS exposure of human blood leukocytes may hamper the inflammatory response to various microbial components. PMID- 19542465 TI - LL-37 complexation with glycosaminoglycans in cystic fibrosis lungs inhibits antimicrobial activity, which can be restored by hypertonic saline. AB - There is an abundance of antimicrobial peptides in cystic fibrosis (CF) lungs. Despite this, individuals with CF are susceptible to microbial colonization and infection. In this study, we investigated the antimicrobial response within the CF lung, focusing on the human cathelicidin LL-37. We demonstrate the presence of the LL-37 precursor, human cathelicidin precursor protein designated 18-kDa cationic antimicrobial protein, in the CF lung along with evidence that it is processed to active LL-37 by proteinase-3. We demonstrate that despite supranormal levels of LL-37, the lung fluid from CF patients exhibits no demonstrable antimicrobial activity. Furthermore Pseudomonas killing by physiological concentrations of exogenous LL-37 is inhibited by CF bronchoalveolar lavage (BAL) fluid due to proteolytic degradation of LL-37 by neutrophil elastase and cathepsin D. The endogenous LL-37 in CF BAL fluid is protected from this proteolysis by interactions with glycosaminoglycans, but while this protects LL-37 from proteolysis it results in inactivation of LL-37 antimicrobial activity. By digesting glycosaminoglycans in CF BAL fluid, endogenous LL-37 is liberated and the antimicrobial properties of CF BAL fluid restored. High sodium concentrations also liberate LL-37 in CF BAL fluid in vitro. This is also seen in vivo in CF sputum where LL-37 is complexed to glycosaminoglycans but is liberated following nebulized hypertonic saline resulting in increased antimicrobial effect. These data suggest glycosaminoglycan LL-37 complexes to be potential therapeutic targets. Factors that disrupt glycosaminoglycan-LL-37 aggregates promote the antimicrobial effects of LL-37 with the caveat that concomitant administration of antiproteases may be needed to protect the now liberated LL-37 from proteolytic cleavage. PMID- 19542466 TI - Cholinergic neural signals to the spleen down-regulate leukocyte trafficking via CD11b. AB - The cholinergic anti-inflammatory pathway is a physiological mechanism that inhibits cytokine production and diminishes tissue injury during inflammation. Recent studies demonstrate that cholinergic signaling reduces adhesion molecule expression and chemokine production by endothelial cells and suppresses leukocyte migration during inflammation. It is unclear how vagus nerve stimulation regulates leukocyte trafficking because the vagus nerve does not innervate endothelial cells. Using mouse models of leukocyte trafficking, we show that the spleen, which is a major point of control for cholinergic modulation of cytokine production, is essential for vagus nerve-mediated regulation of neutrophil activation and migration. Administration of nicotine, a pharmacologic agonist of the cholinergic anti-inflammatory pathway, significantly reduces levels of CD11b, a beta(2)-integrin involved in cell adhesion and leukocyte chemotaxis, on the surface of neutrophils in a dose-dependent manner and this function requires the spleen. Similarly, vagus nerve stimulation significantly attenuates neutrophil surface CD11b levels only in the presence of an intact and innervated spleen. Further mechanistic studies reveal that nicotine suppresses F-actin polymerization, the rate-limiting step for CD11b surface expression. These studies demonstrate that modulation of leukocyte trafficking via cholinergic signaling to the spleen is a specific, centralized neural pathway positioned to suppress the excessive accumulation of neutrophils at inflammatory sites. Activating this mechanism may have important therapeutic potential for preventing tissue injury during inflammation. PMID- 19542467 TI - Major role of gamma delta T cells in the generation of IL-17+ uveitogenic T cells. AB - We show that in vitro activation of interphotoreceptor retinoid-binding protein (IRBP)-specific T cells from C57BL/6 mice immunized with an uveitogenic IRBP peptide (IRBP(1-20)) under TH17-polarizing conditions is associated with increased expansion of T cells expressing the gammadelta TCR. We also show that highly purified alphabeta or gammadelta T cells from C57BL/6 mice immunized with IRBP(1-20) produced only small amounts of IL-17 after exposure to the immunizing Ag in vitro, whereas a mixture of the same T cells produced greatly increased amounts of IL-17. IRBP-induced T cells from IRBP-immunized TCR-delta(-/-) mice on the C57BL/6 genetic background produced significantly lower amounts of IL-17 than did wild-type C57BL/6 mice and had significantly decreased experimental autoimmune uveitis-inducing ability. However, reconstitution of the TCR-delta(-/ ) mice before immunization with a small number of gammadelta T cells from IRBP immunized C57BL/6 mice restored the disease-inducing capability of their IRBP specific T cells and greatly enhanced the generation of IL-17(+) T cells in the recipient mice. Our study suggests that gammadelta T cells are important in the generation and activation of IL-17-producing autoreactive T cells and play a major role in the pathogenesis of experimental autoimmune uveitis. PMID- 19542468 TI - IRAK4 kinase activity is required for Th17 differentiation and Th17-mediated disease. AB - Both IL-23- and IL-1-mediated signaling pathways play important roles in Th17 cell differentiation, cytokine production, and autoimmune diseases. The IL-1R associated kinase 4 (IRAK4) is critical for IL-1/TLR signaling. We show here that inactivation of IRAK4 kinase in mice (IRAK4 KI) results in significant resistance to experimental autoimmune encephalomyelitis due to a reduction in infiltrating inflammatory cells into the CNS and reduced Ag-specific CD4(+) T cell-mediated IL 17 production. Adoptive transfer of myelin oligodendrocyte glycoprotein 35-55 specific IRAK4 KI Th17 cells failed to induce experimental autoimmune encephalomyelitis in either wild-type or IRAK4 KI recipient mice, indicating the lack of autoantigen-specific Th17 cell activities in the absence of IRAK4 kinase activity. Furthermore, the absence of IRAK4 kinase activity blocked induction of IL-23R expression, STAT3 activation by IL-23, and Th17 cytokine expression in differentiated Th17 cells. Importantly, blockade of IL-1 signaling by IL-1RA inhibited Th17 differentiation and IL-23-induced cytokine expression in differentiated Th17 cells. The results of these studies demonstrate that IL-1 mediated IRAK4 kinase activity in T cells is essential for induction of IL-23R expression, Th17 differentiation, and autoimmune disease. PMID- 19542470 TI - Pleiotropic roles of S100A12 in coronary atherosclerotic plaque formation and rupture. AB - Macrophages, cytokines, and matrix metalloproteinases (MMP) play important roles in atherogenesis. The Ca(2+)-binding protein S100A12 regulates monocyte migration and may contribute to atherosclerosis by inducing proinflammatory cytokines in macrophages. We found significantly higher S100A12 levels in sera from patients with coronary artery disease than controls and levels correlated positively with C-reactive protein. S100A12 was released into the coronary circulation from ruptured plaque in acute coronary syndrome, and after mechanical disruption by percutaneous coronary intervention in stable coronary artery disease. In contrast to earlier studies, S100A12 did not stimulate proinflammatory cytokine production by human monocytes or macrophages. Similarly, no induction of MMP genes was found in macrophages stimulated with S100A12. Because S100A12 binds Zn(2+), we studied some functional aspects that could modulate atherogenesis. S100A12 formed a hexamer in the presence of Zn(2+); a novel Ab was generated that specifically recognized this complex. By chelating Zn(2+), S100A12 significantly inhibited MMP 2, MMP-9, and MMP-3, and the Zn(2+)-induced S100A12 complex colocalized with these in foam cells in human atheroma. S100A12 may represent a new marker of this disease and may protect advanced atherosclerotic lesions from rupture by inhibiting excessive MMP-2 and MMP-9 activities by sequestering Zn(2+). PMID- 19542469 TI - Differential regulation of P2X7 receptor activation by extracellular nicotinamide adenine dinucleotide and ecto-ADP-ribosyltransferases in murine macrophages and T cells. AB - Extracellular NAD induces the ATP-independent activation of the ionotropic P2X(7) purinergic receptor (P2X(7)R) in murine T lymphocytes via a novel covalent pathway involving ADP-ribosylation of arginine residues on the P2X(7)R ectodomain. This modification is catalyzed by ART2.2, a GPI-anchored ADP ribosyltransferase (ART) that is constitutively expressed in murine T cells. We previously reported that ART2.1, a related ecto-ART, is up-regulated in inflammatory murine macrophages that constitutively express P2X(7)R. Thus, we tested the hypothesis that extracellular NAD acts via ART2.1 to regulate P2X(7)R function in murine macrophages. Coexpression of the cloned murine P2X(7)R with ART2.1 or ART2.2 in HEK293 cells verified that P2X(7)R is an equivalent substrate for ADP-ribosylation by either ART2.1 or ART2.2. However, in contrast with T cells, the stimulation of macrophages or HEK293 cells with NAD alone did not activate the P2X(7)R. Rather, NAD potentiated ATP-dependent P2X(7)R activation as indicated by a left shift in the ATP dose-response relationship. Thus, extracellular NAD regulates the P2X(7)R in both macrophages and T cells but via distinct mechanisms. Although ADP-ribosylation is sufficient to gate a P2X(7)R channel opening in T cells, this P2X(7)R modification in macrophages does not gate the channel but decreases the threshold for gating in response to ATP binding. These findings indicate that extracellular NAD and ATP can act synergistically to regulate P2X(7)R signaling in murine macrophages and also suggest that the cellular context in which P2X(7)R signaling occurs differs between myeloid and lymphoid leukocytes. PMID- 19542471 TI - Pulmonary eosinophils and their role in immunopathologic responses to formalin inactivated pneumonia virus of mice. AB - Enhanced disease is the term used to describe the aberrant Th2-skewed responses to naturally acquired human respiratory syncytial virus (hRSV) infection observed in individuals vaccinated with formalin-inactivated viral Ags. Here we explore this paradigm with pneumonia virus of mice (PVM), a pathogen that faithfully reproduces features of severe hRSV infection in a rodent host. We demonstrate that PVM infection in mice vaccinated with formalin-inactivated Ags from PVM infected cells (PVM Ags) yields Th2-skewed hypersensitivity, analogous to that observed in response to hRSV. Specifically, we detect elevated levels of IL-4, IL 5, IL-13, and eosinophils in bronchoalveolar lavage fluid of PVM-infected mice that were vaccinated with PVM Ags, but not among mice vaccinated with formalin inactivated Ags from uninfected cells (control Ags). Interestingly, infection in PVM Ag-vaccinated mice was associated with a approximately 10-fold reduction in lung virus titer and protection against weight loss when compared with infected mice vaccinated with control Ags, despite the absence of serum-neutralizing Abs. Given recent findings documenting a role for eosinophils in promoting clearance of hRSV in vivo, we explored the role of eosinophils in altering the pathogenesis of disease with eosinophil-deficient mice. We found that eosinophil deficiency had no impact on virus titer in PVM Ag-vaccinated mice, nor on weight loss or levels of CCL11 (eotaxin-1), IFN-gamma, IL-5, or IL-13 in bronchoalveolar lavage fluid. However, levels of both IL-4 and CCL3 (macrophage inflammatory protein 1alpha) in bronchoalveolar lavage fluid were markedly diminished in PVM Ag vaccinated, PVM-infected eosinophil-deficient mice when compared with wild-type controls. PMID- 19542473 TI - Vaccine-induced, simian immunodeficiency virus-specific CD8+ T cells reduce virus replication but do not protect from simian immunodeficiency virus disease progression. AB - Our limited understanding of the interaction between primate lentiviruses and the host immune system complicates the design of an effective HIV/AIDS vaccine. To identify immunological correlates of protection from SIV disease progression, we immunized two groups of five rhesus macaques (RMs) with either modified vaccinia Ankara (MVA) or MVADeltaudg vectors that expressed SIVmac239 Gag and Tat. Both vectors raised a SIV-specific CD8(+) T cell response, with a magnitude that was greater in mucosal tissues than in peripheral blood. After challenge with SIVmac239, all vaccinated RMs showed mucosal and systemic CD8(+) T cell recall responses that appeared faster and were of greater magnitude than those in five unvaccinated control animals. All vaccinated RMs showed a approximately 1-log lower peak and early set-point SIV viral load than the unvaccinated animals, and then, by 8 wk postchallenge, exhibited levels of viremia similar to the controls. We observed a significant direct correlation between the magnitude of postchallenge SIV-specific CD8(+) T cell responses and SIV viral load. However, vaccinated RMs showed no protection from either systemic or mucosal CD4(+) T cell depletion and no improved survival. The observation that vaccine-induced, SIV specific CD8(+) T cells that partially control SIVmac239 virus replication fail to protect from immunological or clinical progression of SIV infection underscores both the complexity of AIDS pathogenesis and the challenges of properly assessing the efficacy of candidate AIDS vaccines. PMID- 19542472 TI - Aberrant tissue localization of fungus-specific CD4+ T cells in IL-10-deficient mice. AB - Aspergillus fumigatus, a common environmental fungus, can cause lethal invasive infections in immunocompromised hosts. In immunocompetent individuals, however, inhaled A. fumigatus spores prime CD4(+) T cells and activate immune responses that prevent invasive infection. Calibration of inflammatory responses to levels that prevent fungal invasion without inducing collateral tissue damage is essential for host survival, but the underlying regulatory mechanisms remain undefined. Although IL-10 is a validated regulatory cytokine that suppresses immune responses, and IL-10 deficiency or blockade generally enhances immune responses, we find that A. fumigatus-specific T cell frequencies are markedly reduced in airways of IL-10-deficient mice. T cell priming, proliferation, and survival were unaffected by IL-10 deficiency and did not account for decreased frequencies of A. fumigatus-specific T cells in the airways of IL-10-deficient mice. Instead, IL-10 deficiency results in redistribution of A. fumigatus specific T cells from infected lungs to the gut, a process that is reversed by antibiotic-mediated depletion of intestinal microbes. Our studies demonstrate that disregulated immune responses in the gut can result in dramatic redistribution of pathogen-specific T cells within the host. PMID- 19542474 TI - Dissociation between memory accuracy and memory confidence following bilateral parietal lesions. AB - Numerous functional neuroimaging studies have observed lateral parietal lobe activation during memory tasks: a surprise to clinicians who have traditionally associated the parietal lobe with spatial attention rather than memory. Recent neuropsychological studies examining episodic recollection after parietal lobe lesions have reported differing results. Performance was preserved in unilateral lesion patients on source memory tasks involving recollecting the context in which stimuli were encountered, and impaired in patients with bilateral parietal lesions on tasks assessing free recall of autobiographical memories. Here, we investigated a number of possible accounts for these differing results. In 3 experiments, patients with bilateral parietal lesions performed as well as controls at source recollection, confirming the previous unilateral lesion results and arguing against an explanation for those results in terms of contralesional compensation. Reducing the behavioral relevance of mnemonic information critical to the source recollection task did not affect performance of the bilateral lesion patients, indicating that the previously observed reduced autobiographical free recall might not be due to impaired bottom-up attention. The bilateral patients did, however, exhibit reduced confidence in their source recollection abilities across the 3 experiments, consistent with a suggestion that parietal lobe lesions might lead to impaired subjective experience of rich episodic recollection. PMID- 19542475 TI - Association between tobacco smoking and active tuberculosis in Taiwan: prospective cohort study. AB - RATIONALE: Previous case-control studies and a small number of cohort studies in high-risk populations have found an association between tobacco and active tuberculosis, but no cohort studies have been conducted in the general population on this association to date. OBJECTIVES: To investigate the association between tobacco smoking and active tuberculosis in a cohort of a general population. METHODS: 17,699 participants (>or=12 y of age) in Taiwan National Health Interview Survey were followed up from 2001 to 2004. Smoking status and other covariates were measured by an in-person interview at baseline. Incident cases of active tuberculosis were identified from the National Health Insurance database. Multivariate logistic regression was used to estimate the association between smoking status and active tuberculosis, with adjustment for age, sex, alcohol consumption, socioeconomic status, and other covariates. MEASUREMENTS AND MAIN RESULTS: Fifty-seven new cases of active tuberculosis occurred during the 3.3 years of follow-up. Current smoking was associated with an increased risk of active tuberculosis (adjusted odds ratio [OR], 1.94; 95% confidence interval, 1.01-3.73). The association was stronger among those less than 65 years of age (adjusted OR, 3.04) than those greater than 65 years of age (adjusted OR, 0.78; P(interaction) = 0.036). We found significant dose-response relations for cigarettes per day (P(trend) = 0.0036), years of smoking (P(trend) = 0.023), and pack-years (P(trend) = 0.0023). CONCLUSIONS: Tobacco smoking was associated with a twofold increased risk of active tuberculosis in a representative cohort of Taiwan's population. PMID- 19542477 TI - Vascular dysfunction in chronic obstructive pulmonary disease. AB - RATIONALE: Cardiovascular disease is a major cause of morbidity and mortality in patients with chronic obstructive pulmonary disease (COPD), which may in part be attributable to abnormalities of systemic vascular function. It is unclear whether such associations relate to the presence of COPD or prior smoking habit. OBJECTIVES: To undertake a comprehensive assessment of vascular function in patients with COPD and healthy control subjects matched for smoking history. METHODS: Eighteen men with COPD were compared with 17 healthy male control subjects matched for age and lifetime cigarette smoke exposure. Participants were free from clinically evident cardiovascular disease. MEASUREMENTS AND MAIN RESULTS: Pulse wave velocity and pulse wave analysis were measured via applanation tonometry at carotid, radial, and femoral arteries. Blood flow was measured in both forearms using venous occlusion plethysmography during intrabrachial infusion of endothelium-dependent vasodilators (bradykinin, 100 1,000 pmol/min; acetylcholine, 5-20 microg/min) and endothelium-independent vasodilators (sodium nitroprusside, 2-8 microg/min; verapamil, 10-100 microg/min). Tissue plasminogen activator (t-PA) was measured in venous plasma before and during bradykinin infusions. Patients with COPD have greater arterial stiffness (pulse wave velocity, 11 +/- 2 vs. 9 +/- 2 m/s; P = 0.003; augmentation index, 27 +/- 10 vs. 21 +/- 6%; P = 0.028), but there were no differences in endothelium-dependent and -independent vasomotor function or bradykinin-induced endothelial t-PA release (P > 0.05 for all). CONCLUSIONS: COPD is associated with increased arterial stiffness independent of cigarette smoke exposure. However, this abnormality is not explained by systemic endothelial dysfunction. Increased arterial stiffness may represent the mechanistic link between COPD and the increased risk for cardiovascular disease associated with this condition. PMID- 19542476 TI - Shortening treatment in adults with noncavitary tuberculosis and 2-month culture conversion. AB - RATIONALE: Cavitary disease and delayed culture conversion have been associated with relapse. Combining patient characteristics and measures of bacteriologic response might allow treatment shortening with current drugs in some patients. OBJECTIVES: To assess whether treatment could be shortened from 6 to 4 months in patients with noncavitary tuberculosis whose sputum cultures converted to negative after 2 months. METHODS: This study was a randomized, open-label equivalence trial. HIV-uninfected adults with noncavitary tuberculosis were treated daily with isoniazid, rifampin, pyrazinamide, and ethambutol for 2 months, followed by 2 months of isoniazid and rifampin. After 4 months, patients with drug-susceptible TB whose sputum cultures on solid media were negative after 8 weeks of treatment were randomly assigned to continue treatment for 2 more months or to stop treatment. Patients were followed for relapse for 30 months after beginning treatment. MEASUREMENTS AND MAIN RESULTS: Enrollment was stopped by the safety monitoring committee after 394 patients were enrolled due to apparent increased risk for relapse in the 4-month arm. A total of 370 patients were eligible for per protocol analysis. Thirteen patients in the 4-month arm relapsed, compared with three subjects in the 6-month arm (7.0 vs. 1.6%; risk difference, 0.054; 95% confidence interval with Hauck-Anderson correction, 0.01 0.10). CONCLUSION: Shortening treatment from 6 to 4 months in adults with noncavitary disease and culture conversion after 2 months using current drugs resulted in a greater relapse rate. The combination of noncavitary disease and 2 month culture conversion was insufficient to identify patients with decreased risk for relapse. PMID- 19542478 TI - Early daycare is associated with an increase in airway symptoms in early childhood but is no protection against asthma or atopy at 8 years. AB - RATIONALE: Daycare exposes young children to more infections early in life and may thereby prevent the development of asthma and allergy. OBJECTIVES: To prospectively study the effect of daycare on the development of asthma and allergic sensitization during the first 8 years of life. METHODS: In the Prevention and Incidence of Asthma and Mite Allergy birth cohort 3,963 newborn children were followed prospectively for 8 years. Daycare use and respiratory health were assessed yearly by questionnaires. At 8 years, sensitization to airborne allergens and airway responsiveness were measured. Daycare was defined as early (aged 0-2 yr), late (aged 2-4 yr), or none (no daycare before age 4 yr). Associations of daycare and/or older siblings with asthma symptoms (wheezing, shortness of breath, and inhaled steroids taken in the last year), airway responsiveness, and allergic sensitization were assessed in a longitudinal repeated-event analysis. MEASUREMENTS AND MAIN RESULTS: Children with early daycare had more wheezing in the first years of life, but less wheezing and steroid use between 4 and 8 years of age. At the age of 8 years, early daycare was not protective for asthma symptoms (adjusted odds ratio [aOR], 0.99; 95% confidence interval [CI], 0.74-1.32), allergic sensitization (aOR 0.86; 95% CI, 0.63-1.18), or airway hyperresponsiveness (aOR, 0.80; 95% CI, 0.57-1.14). The transient reduction in airway symptoms between age 4 and 8 years was only observed in children without older siblings. CONCLUSION: Early daycare is associated with an increase in airway symptoms until the age of 4 years, and fewer symptoms between the ages of 4 and 8 years. We found no protection against asthma symptoms, hyperresponsiveness, or allergic sensitization at the age of 8 years. PMID- 19542479 TI - Inhomogeneity of lung parenchyma during the open lung strategy: a computed tomography scan study. AB - RATIONALE: The open lung strategy aims at reopening (recruitment) of nonaerated lung areas in patients with acute respiratory distress syndrome, avoiding tidal alveolar hyperinflation in the limited area of normally aerated tissue (baby lung). OBJECTIVES: We tested the hypothesis that recruited lung areas do not resume elastic properties of adjacent baby lung. METHODS: Twenty-five anesthetized, mechanically ventilated pigs were studied. Four lung-healthy pigs served as controls and the remaining 21 were divided into three groups (n = 7 each) in which lung injury was produced by surfactant lavage, lipopolysaccharide infusion, or hydrochloride inhalation. Computed tomography scans, respiratory mechanics, and gas exchange parameters were recorded under three conditions: at baseline, during lung recruitment maneuver, and at end-expiration and end inspiration when ventilating after an open lung protocol. MEASUREMENTS AND MAIN RESULTS: During recruitment maneuver and open lung protocol, the gas volume entering the insufficiently aerated compartment was 96% (75-117%) and 48% (41 63%) (median [interquartile range]) of the functional residual capacity measured before and at zero end-expiratory pressure, respectively. Nonetheless, the volume of hyperinflated lung increased during both recruitment maneuver (by 1-28% of total lung volume; P < 0.01) and open lung protocol ventilation at end inspiration (by 1-15% of total lung volume; P < 0.01). Regional elastance of recruited lung tissue was consistently higher than that of the baby lung regardless of the ARDS model (P < 0.01). CONCLUSIONS: Alveolar recruitment is not protective against hyperinflation of the baby lung because lung parenchyma is inhomogeneous during ventilation with the open lung strategy. PMID- 19542481 TI - Daily physical activity in patients with chronic obstructive pulmonary disease is mainly associated with dynamic hyperinflation. AB - RATIONALE: Although the major limitation to exercise performance in patients with COPD is dynamic hyperinflation, little is known about its relation to daily physical activity. OBJECTIVES: To analyze the contribution of dynamic hyperinflation, exercise tolerance, and airway oxidative stress to physical activity in patients with COPD. METHODS: In a cross-sectional study, we included 110 patients with moderate to very severe COPD. Daily physical activity was measured using a triaxial accelerometer providing a mean of 1-minute movement epochs as vector magnitude units (VMU). Patients performed the 6-minute walk test, incremental exercise test with measurement of breathing pattern and operating lung volumes, and constant-work rate test at 75% of maximal work rate. MEASUREMENTS AND MAIN RESULTS: Using the GOLD stage and BODE index, we determined arterial blood gases, lung volumes, diffusing capacity, and biomarkers in exhaled breath condensate. Daily physical activity was lower in the 89 patients who developed dynamic hyperinflation than in the 21 who did not (n =161 [SD 70] vs. n = 288 [SD 85] VMU; P = 0.001). Physical activity was mainly related to distance walked in 6 minutes (r = 0.72; P = 0.001), Vo(2) (r = 0.63; P = 0.001), change in end-expiratory lung volume during exercise (r = -0.73; P = 0.001), endurance time (r = 0.61; P = 0.001), and 8-isoprostane in exhaled breath condensate (r = -0.67; P = 0.001). In a multivariate linear regression analysis using VMU as a dependent variable, dynamic hyperinflation, change in end-expiratory lung volume, and distance walked in 6 minutes were retained in the prediction model (r(2) = 0.84; P = 0.001). CONCLUSIONS: Daily physical activity of patients with COPD is mainly associated with dynamic hyperinflation, regardless of severity classification. PMID- 19542480 TI - Cigarette smoking is associated with subclinical parenchymal lung disease: the Multi-Ethnic Study of Atherosclerosis (MESA)-lung study. AB - RATIONALE: Cigarette smoking is a risk factor for diffuse parenchymal lung disease. Risk factors for subclinical parenchymal lung disease have not been described. OBJECTIVES: To determine if cigarette smoking is associated with subclinical parenchymal lung disease, as measured by spirometric restriction and regions of high attenuation on computed tomography (CT) imaging. METHODS: We examined 2,563 adults without airflow obstruction or clinical cardiovascular disease in the Multi-Ethnic Study of Atherosclerosis, a population-based cohort sampled from six communities in the United States. Cumulative and current cigarette smoking were assessed by pack-years and urine cotinine, respectively. Spirometric restriction was defined as a forced vital capacity less than the lower limit of normal. High attenuation areas on the lung fields of cardiac CT scans were defined as regions having an attenuation between -600 and -250 Hounsfield units, reflecting ground-glass and reticular abnormalities. Generalized additive models were used to adjust for age, gender, race/ethnicity, smoking status, anthropometrics, center, and CT scan parameters. MEASUREMENTS AND MAIN RESULTS: The prevalence of spirometric restriction was 10.0% (95% confidence interval [CI], 8.9-11.2%) and increased relatively by 8% (95% CI, 3-12%) for each 10 cigarette pack-years in multivariate analysis. The median volume of high attenuation areas was 119 cm(3) (interquartile range, 100-143 cm(3)). The volume of high attenuation areas increased by 1.6 cm(3) (95% CI, 0.9-2.4 cm(3)) for each 10 cigarette pack-years in multivariate analysis. CONCLUSIONS: Smoking may cause subclinical parenchymal lung disease detectable by spirometry and CT imaging, even among a generally healthy cohort. PMID- 19542482 TI - Cholinergic receptor signaling modulates spontaneous firing of sinoatrial nodal cells via integrated effects on PKA-dependent Ca(2+) cycling and I(KACh). AB - Prior studies indicate that cholinergic receptor (ChR) activation is linked to beating rate reduction (BRR) in sinoatrial nodal cells (SANC) via 1) a G(i) coupled reduction in adenylyl cyclase (AC) activity, leading to a reduction of cAMP or protein kinase A (PKA) modulation of hyperpolarization-activated current (I(f)) or L-type Ca(2+) currents (I(Ca,L)), respectively; and 2) direct G(i) coupled activation of ACh-activated potassium current (I(KACh)). More recent studies, however, have indicated that Ca(2+) cycling by the sarcoplasmic reticulum within SANC (referred to as a Ca(2+) clock) generates rhythmic, spontaneous local Ca(2+) releases (LCR) that are AC-PKA dependent. LCRs activate Na(+)-Ca(2+) exchange (NCX) current, which ignites the surface membrane ion channels to effect an AP. The purpose of the present study was to determine how ChR signaling initiated by a cholinergic agonist, carbachol (CCh), affects AC, cAMP, and PKA or sarcolemmal ion channels and LCRs and how these effects become integrated to generate the net response to a given intensity of ChR stimulation in single, isolated rabbit SANC. The threshold CCh concentration ([CCh]) for BRR was approximately 10 nM, half maximal inhibition (IC(50)) was achieved at 100 nM, and 1,000 nM stopped spontaneous beating. G(i) inhibition by pertussis toxin blocked all CCh effects on BRR. Using specific ion channel blockers, we established that I(f) blockade did not affect BRR at any [CCh] and that I(KACh) activation, evidenced by hyperpolarization, first became apparent at [CCh] > 30 nM. At IC(50), CCh reduced cAMP and reduced PKA-dependent phospholamban (PLB) phosphorylation by approximately 50%. The dose response of BRR to CCh in the presence of I(KACh) blockade by a specific inhibitor, tertiapin Q, mirrored that of CCh to reduced PLB phosphorylation. At IC(50), CCh caused a time-dependent reduction in the number and size of LCRs and a time dependent increase in LCR period that paralleled coincident BRR. The phosphatase inhibitor calyculin A reversed the effect of IC(50) CCh on SANC LCRs and BRR. Numerical model simulations demonstrated that Ca(2+) cycling is integrated into the cholinergic modulation of BRR via LCR-induced activation of NCX current, providing theoretical support for the experimental findings. Thus ChR stimulation-induced BRR is entirely dependent on G(i) activation and the extent of G(i) coupling to Ca(2+) cycling via PKA signaling or to I(KACh): at low [CCh], I(KACh) activation is not evident and BRR is attributable to a suppression of cAMP-mediated, PKA dependent Ca(2+) signaling; as [CCh] increases beyond 30 nM, a tight coupling between suppression of PKA-dependent Ca(2+) signaling and I(KACh) activation underlies a more pronounced BRR. PMID- 19542483 TI - Differential structural and functional changes in penile and coronary arteries from obese Zucker rats. AB - Erectile dysfunction frequently coexists with coronary artery disease and has been proposed as a potential marker for silent coronary artery disease in type 2 diabetes. In the present study, we comparatively assessed the structural and functional changes of both penile arteries (PAs) and coronary arteries (CAs) from a prediabetic animal model. PAs and CAs from 17- to 18-wk-old obese Zucker rats (OZRs) and from their control counterparts [lean Zucker rats (LZRs)] were mounted in microvascular myographs to evaluate vascular function, and stained arteries were subjected to morphometric analysis. Endothelial nitric oxide (NO) synthase (eNOS) protein expression was also assessed. The internal diameter was reduced and the wall-to-lumen ratio was increased in PAs from OZRs, but structure was preserved in CAs. ACh-elicited relaxations were severely impaired in PAs but not in CAs from OZRs, although eNOS expression was unaltered. Contractions to norepinephrine and 5-HT were significantly enhanced in both PAs and CAs, respectively, from OZRs. Blockade of NOS abolished endothelium-dependent relaxations in PAs and CAs and potentiated norepinephrine and 5-HT contractions in arteries from LZRs but not from OZRs. The vasodilator response to the phosphodiesterase 5 inhibitor sildenafil was reduced in both PAs and CAs from OZRs. Pretreatment with SOD reduced the enhanced vasoconstriction in both PAs and CAs from OZRs but did not restore ACh-induced relaxations in PAs. In conclusion, the present results demonstrate vascular inward remodeling in PAs and a differential impairment of endothelial relaxant responses in PAs and CAs from insulin-resistant OZRs. Enhanced superoxide production and reduced basal NO activity seem to underlie the augmented vasoconstriction in both PAs and CAs. The severity of the structural and functional abnormalities in PAs might anticipate the vascular dysfunction of the more preserved coronary vascular bed. PMID- 19542484 TI - Phenylephrine and sustained acidosis activate the neonatal rat cardiomyocyte Na+/H+ exchanger through phosphorylation of amino acids Ser770 and Ser771. AB - The mammalian Na(+)/H(+) exchanger isoform 1 (NHE1) is a ubiquitously expressed membrane protein that regulates intracellular pH in the myocardium. NHE1 is also important in mediating myocardial hypertrophy, and the blockage of NHE1 activity prevents hypertrophy and reduces ischemia-reperfusion injury in animal models. We recently demonstrated that extracellular-regulated kinase (ERK)-mediated activation of NHE1 occurs during ischemia-reperfusion of the myocardium. To understand the regulation of NHE1 in the myocardium by phosphorylation, we expressed a series of adenoviruses that express wild-type and mutant cDNA for NHE1. All exogenous cDNA for NHE1 had additional mutations [Leu(163)Phe/Gly(174)Ser], which increases NHE1 resistance to EMD-87580 (a specific blocker of NHE1) 100-fold, and allowed the measurement of exogenous NHE1 while inhibiting endogenous NHE1. By examining the effects of a series of mutations of the NHE1 cytosolic region, we determined that the amino acids Ser(770) and Ser(771) were essential for the acute activation of NHE1 activity in rat cardiomyocytes. The specific mutation of either residue prevented the rapid activation of exchanger activity by a sustained intracellular acidosis through ERK-dependent pathways. The same amino acids were critical to phenylephrine mediated, ERK-dependent activation of NHE1 activity and increased the phosphorylation in intact rat cardiomyocytes. The results demonstrate that both sustained intracellular acidosis and phenylephrine rapidly activate the NHE1 protein in intact cardiac cells through ERK-dependent pathways that act on a common pathway mediated by amino acids Ser(770) and Ser(771) of the cytosolic tail of the protein. PMID- 19542485 TI - Lysozyme, a mediator of sepsis that intrinsically generates hydrogen peroxide to cause cardiovascular dysfunction. AB - In septic shock, cardiovascular collapse is caused by the release of inflammatory mediators. We previously found that lysozyme (Lzm-S), released from leukocytes, contributed to the myocardial depression and arterial vasodilation that develop in canine models of septic shock. To cause vasodilation, Lzm-S generates hydrogen peroxide (H(2)O(2)) that activates the smooth muscle soluble guanylate cyclase (sGC) pathway, although the mechanism of H(2)O(2) generation is not known. To cause myocardial depression, Lzm-S binds to the endocardial endothelium, resulting in the formation of nitric oxide (NO) and subsequent activation of myocardial sGC, although the initial signaling event is not clear. In this study, we examined whether the myocardial depression produced by Lzm-S was also caused by the generation of H(2)O(2) and whether Lzm-S could intrinsically generate H(2)O(2) as has been described for other protein types. In a canine ventricular trabecular preparation, we found that the peroxidizing agent Aspergillus niger catalase, that would breakdown H(2)O(2), prevented Lzm-S- induced decrease in contraction. We also found that compound I, a species of catalase formed during H(2)O(2) metabolism, could contribute to the NO generation caused by Lzm-S. In tissue-free experiments, we used a fluorometric assay (Ultra Amplex red H(2)O(2) assay) and electrochemical sensor techniques, respectively, to measure H(2)O(2) generation. We found that Lzm-S could generate H(2)O(2) and, furthermore, that this generation could be attenuated by the singlet oxygen quencher sodium azide. This study shows that Lzm-S, a mediator of sepsis, is able to intrinsically generate H(2)O(2). Moreover, this generation may activate H(2)O(2)-dependent pathways leading to cardiovascular collapse in septic shock. PMID- 19542486 TI - The arrythmogenic potential of post-myocardial infarction cytokine treatment. PMID- 19542488 TI - Dose-dependent activation of distinct hypertrophic pathways by serotonin in cardiac cells. AB - There is substantial evidence supporting a hypertrophic action of serotonin [5 hydroxytryptamine (5-HT)] in cardiomyocytes. However, little is known about the mechanisms involved. We previously demonstrated that 5-HT-induced hypertrophy depends, in part, on the generation of reactive oxygen species by monoamine oxidase-A (MAO-A) (see Ref. 3). Cardiomyocytes express 5-HT(2) receptors, which may also participate in hypertrophy. Here, we analyzed the respective contribution of 5-HT(2) receptors and MAO-A in H9C2 cardiomyoblast hypertrophy. 5 HT induced a dose-dependent increase in [(3)H]leucine incorporation and stimulation of two markers of cardiac hypertrophy, ANF-luc and alphaSK-actin-luc reporter genes. Experiments using 1 microM 5-HT showed that hypertrophic response occurred independently from MAO-A. Using pharmacological inhibitors (M100907 and ketanserin), we identified a novel mechanism of action involving 5-HT(2A) receptors and requiring Ca(2+)/calcineurin/nuclear factor of activated T-cell activation. The activation of this hypertrophic pathway was fully prevented by 5 HT(2A) inhibitors and was unaffected by MAO inhibition. When 10 microM 5-HT was used, an additional hypertrophic response, prevented by the MAO inhibitors pargyline and RO 41-1049, was observed. Unlike the 5-HT(2A)-receptor-mediated H9C2 cell hypertrophy, MAO-A-dependent hypertrophic response required activation of extracellular-regulated kinases. In conclusion, our results show the existence of a dose-dependent shift of activation of distinct intracellular pathways involved in 5-HT-mediated hypertrophy of cardiac cells. PMID- 19542487 TI - Proliferating cardiac microtubules. PMID- 19542489 TI - Mechanical stress analysis of a rigid inclusion in distensible material: a model of atherosclerotic calcification and plaque vulnerability. AB - The role of atherosclerotic calcification in plaque rupture remains controversial. In previous analyses using finite element model analysis, circumferential stress was reduced by the inclusion of a calcium deposit in a representative human anatomical configuration. However, a recent report, also using finite element analysis, suggests that microscopic calcium deposits increase plaque stress. We used mathematical models to predict the effects of rigid and liquid inclusions (modeling a calcium deposit and a lipid necrotic core, respectively) in a distensible material (artery wall) on mechanical failure under uniaxial and biaxial loading in a range of configurations. Without inclusions, stress levels were low and uniform. In the analytical model, peak stresses were elevated at the edges of a rigid inclusion. In the finite element model, peak stresses were elevated at the edges of both inclusions, with minimal sensitivity to the wall distensibility and the size and shape of the inclusion. Presence of both a rigid and a soft inclusion enlarged the region of increased wall stress compared with either alone. In some configurations, the rigid inclusion reduced peak stress at the edge of the soft inclusion but simultaneously increased peak stress at the edge of the rigid inclusion and increased the size of the region affected. These findings suggest that the presence of a calcium deposit creates local increases in failure stress, and, depending on relative position to any neighboring lipid pools, it may increase peak stress and the plaque area at risk of mechanical failure. PMID- 19542490 TI - Hypoxia induces B-type natriuretic peptide release in cell lines derived from human cardiomyocytes. AB - B-type natriuretic peptide (BNP) is a peptide hormone of myocardial origin with significant cardioprotective properties. Patients with myocardial ischemia present with high levels of BNP in plasma and elevated expression in the myocardium. However, the molecular mechanisms of BNP induction in the ischemic myocardium are not well understood. The aim of the investigation was to assess whether myocardial hypoxia induces the production of BNP in human ventricular myocytes. To test the hypothesis that reduced oxygen tension can directly stimulate BNP gene expression and release in the absence of hemodynamic or neurohormonal stimuli, we used an in vitro model system of cultured human ventricular myocytes (AC16 cells). Cells were cultured under normoxic (21% O(2)) or hypoxic (5% O(2)) conditions for up to 48 h. The accumulation of BNP, atrial natriuretic peptide (ANP), and vascular endothelial growth factor (VEGF) was then measured. Hypoxia stimulated the protein release of BNP and VEGF but not ANP. In concordance, the increased mRNA levels of BNP and VEGF but not ANP were found on culturing AC16 cells under hypoxic conditions. The analysis of the transcriptional activity of the hypoxia-inducible factor 1 (HIF-1) in nuclear extracts showed that HIF-1 activity was induced under hypoxic conditions. Finally, the treatment of AC16 cells with the HIF-1 inhibitor rotenone in hypoxia inhibited BNP and VEGF release. In conclusion, these data indicate that hypoxia induces the synthesis and secretion of BNP in human ventricular myocytes, likely through HIF-1-enhanced transcriptional activity. PMID- 19542492 TI - Temporal changes in vascular reactivity in early diabetes mellitus in rats: role of changes in endothelial factors and in phosphodiesterase activity. AB - The aims of this study were to study the influence of the duration of diabetes, the role of endothelial-derived vasodilators, and the role of phosphodiesterase (PDE) isoform activity in the early changes in vascular reactivity of aortic rings from diabetic rats. Diabetes mellitus was induced in female rats by intravenous streptozotocin (85 mg/kg). Two or 4 wk later, thoracic aortic rings from control and diabetic rats were isolated, and vascular responses to acetylcholine (ACh), S-nitroso-N-acetylpenicillamine (SNAP) [nitric oxide (NO) donor], DMPPO (PDE5 inhibitor), and phenylephrine (PE) were obtained in the presence and absence of endothelium or other drugs. PDE isoform activity was also measured. At 2 wk, responses to ACh and DMPPO were enhanced, whereas those to PE were attenuated in diabetic rats relative to controls. Indomethacin and SQ-29548 (a thromboxane A(2) receptor antagonist), but not N(G)-nitro-L-arginine methyl ester, corrected these differences. The responses to SNAP, and cAMP and cGMP hydrolytic activities, were similar in the two groups. In contrast, at 4 wk, ACh, DMPPO, and PE produced similar responses in the two groups: N(G)-nitro-L-arginine methyl ester rendered the response to PE lower in the diabetic group, and this was corrected by indomethacin, but not SQ-29548, treatment. The response to SNAP was greater in the diabetic group, and this was corrected by DMPPO. Activity of all PDEs was decreased at 4 wk. We conclude that, at 2 wk, there is modulation of thromboxane A(2) production, but no change in the NO system or PDE isoform activities. At 4 wk, a reduction in NO activity is superimposed; at this stage, PDE activity is reduced, together with increased production of vasodilating prostaglandins, possibly as a compensatory mechanism to maintain normal vascular reactivity. PMID- 19542491 TI - Abl knockout differentially affects p130 Crk-associated substrate, vinculin, and paxillin in blood vessels of mice. AB - Actin polymerization has recently emerged as an important cellular process that regulates smooth muscle contraction. Abelson tyrosine kinase (Abl) has been implicated in the regulation of actin dynamics and force development in vascular smooth muscle. In the present study, the systolic blood pressure was lower in Abl(-/-) knockout mice compared with wild-type mice. The knockout of Abl diminished the tyrosine phosphorylation of p130 Crk-associated substrate (CAS, an adapter protein associated with smooth muscle contraction) in resistance arteries upon stimulation with phenylephrine or angiotensin II. The agonist-elicited enhancement of F-actin-to-G-actin ratios in arteries assessed by fluorescent microscopy was also reduced in Abl(-/-) mice. It has been known that vinculin is a structural protein that links actin filaments to extracellular matrix via transmembrane integrins, whereas paxillin is a signaling protein associated with focal contacts mediating actin cytoskeleton remodeling. The expression of vinculin and paxillin at protein and messenger levels was lower in arterial vessels from Abl knockout mice. However, the agonist-induced increase in myosin phosphorylation was not attenuated in arteries from Abl knockout mice. These results indicate that Abl differentially regulates Crk-associated substrate, vinculin, and paxillin in arterial vessels. The Abl-regulated cellular process and blood pressure are independent of myosin activation in vascular smooth muscle. PMID- 19542494 TI - Promoting colorectal cancer screening: which interventions work? PMID- 19542493 TI - Epigenetic profiling at mouse imprinted gene clusters reveals novel epigenetic and genetic features at differentially methylated regions. AB - Genomic imprinting arises from allele-specific epigenetic modifications that are established during gametogenesis and that are maintained throughout somatic development. These parental-specific modifications include DNA methylation and post-translational modifications to histones, which create allele-specific active and repressive domains at imprinted regions. Through the use of a high-density genomic tiling array, we generated DNA and histone methylation profiles at 11 imprinted gene clusters in the mouse from DNA and from chromatin immunoprecipitated from sperm, heart, and cerebellum. Our analysis revealed that despite high levels of differential DNA methylation at non-CpG islands within these regions, imprinting control regions (ICRs) and secondary differentially methylated regions (DMRs) were identified by an overlapping pattern of H3K4 trimethylation (active chromatin) and H3K9 trimethylation (repressive chromatin) modifications in somatic tissue, and a sperm differentially methylated region (sDMR; sperm not equal somatic tissue). Using these features as a common signature of DMRs, we identified 11 unique regions that mapped to known imprinted genes, to uncharacterized genes, and to intergenic regions flanking known imprinted genes. A common feature among these regions was the presence of a CpG island and an array of tandem repeats. Collectively, this study provides a comprehensive analysis of DNA methylation and histone H3K4me3 and H3K9me3 modifications at imprinted gene clusters, and identifies common epigenetic and genetic features of regions regulating genomic imprinting. PMID- 19542495 TI - End-of-life care: Communication and a stable patient-physician relationship lead to better decisions. PMID- 19542496 TI - Use of crossbreeding with beef bulls in dairy herds: effect on age, body weight, price, and market value of calves sold at livestock auctions. AB - The aim of this study was to investigate the effect of different breeds and breed crosses on age (AC, d), BW (kg), price (PR, $/kg), and market value (MV, $/calf) of purebred and crossbred calves sold for veal and beef production. The Kovieh wholesale cattle organization (Bolzano, Italy) grouped calves from several dairy herds located in the Trentino-Sudtirol region in Italy and sold them by public auctions. Data on AC, BW, PR, and MV from 96,458 calves were recorded from January 2003 to December 2007 and consisted of 4 pure breeds [2 dairy, Brown Swiss (BS) and Holstein-Friesian (HF); and 2 dual-purpose, Simmental (SI) and Alpine Grey (AG)], and 8 crossbreds by crosses of Limousin (LI) and Belgian Blue (BB) with the 4 dam breeds. Least squares means for AC, BW, PR, and MV were calculated for breeds and breed crosses with a model that included fixed effects of herd of birth, age (except for AC), sex, and breed of the calf, year and season of auction, and interactions between the main effects. The coefficients of determination of the models were 0.41, 0.51, 0.84, and 0.82 for AC, BW, PR, and MV, respectively. Sex, age, and breed were the most relevant sources of variation for BW (P < 0.001), whereas breed and sex were the most important sources of variation for AC, PR, and MV (P < 0.001). Also, PR and MV were significantly influenced (P < 0.01) by all the effects included in the model, except for season x age interaction in the case of MV. Market value of male was greater (P < 0.001) than that of female calves, with the exception of BS (-$28.76/calf) and HF ( $20.70/calf) purebred males. Dual-purpose purebred calves presented greater (P < 0.001) PR and MV than dairy purebreds (MV of $426.97/calf and $307.96/calf for SI and AG, and $256.24/calf and $275.65/calf for BS and HF, respectively). Calves from SI and AG dams had greater (P < 0.001) BW, PR, and MV than calves from BS and HF dams. Calves from SI cows had greater (P < 0.001) BW, PR, and MV than calves from AG cows. Crossbreeding with beef bulls increased (P < 0.001) BW, PR, and MV of calves from dairy and dual-purpose dams. Crossbreeding with BB bulls increased PR (+$2.58 +/- 0.04/kg; P < 0.001) and MV (+$190.84 +/- 3.62/calf; P < 0.001) of calves much more than LI. The use of beef bull semen on dairy herds resulted in an economic revenue from selling crossbred calves. PMID- 19542497 TI - Effect of blood lactate-guided conditioning of horses with exercises of differing durations and intensities on heart rate and biochemical blood variables. AB - The velocity at which blood lactate concentration ([LA]) of 4 mmol/L is reached (v(4)) is widely used to determine fitness, but there are few published data on using [LA] as a guide for the exercise speed for training in horses. In this study, the effect of 3 conditioning programs with [LA] guided exercise speeds on v(4), v(200) (speed at a heart rate of 200 beats/min), blood [LA], plasma FFA ([FFA]), and alanine concentrations ([alanine]), before and after exercise, as well as heart rate during exercise, of horses was examined. Six 2-yr-old Haflinger stallions underwent an initial treadmill-based standard exercise test (SET). A regression analysis [LA]-speed relationship was used to calculate v(1.5), v(2.5), and v(4). Horses were then randomly assigned to 1 of 3 conditioning programs according to a 3 x 6 Latin square design. During 6 wk, horses exercised on the treadmill every other day for 45 min at their calculated v(1.5) or v(2.5) or 25 min at their v(4). Each conditioning period (CDP) was followed by 5 wk without conditioning. At 2 and 9 d, and 5 wk, after the end of the CDP, the horses performed another SET to evaluate again the v(4) and v(200). Blood [LA], plasma [FFA], and [alanine] were measured before and after heart rate during exercise sessions 1, 11, and 21 in each CDP. None of the exercise programs had an effect on v(4) and v(200) (P > 0.05). The increase of the mean [LA] after exercise decreased during CDP (P < 0.05), and the increase of mean heart rate during exercise tended to decrease as well (P = 0.07). There was no difference among the conditioning programs. Plasma [FFA] before exercise was not influenced by the CDP (P > 0.05). The plasma [FFA] was always greater after exercise (P < 0.05), but there was no difference among conditioning programs. Overall, the increase was greatest after the 21st exercise session compared with the 1st and 11th exercise sessions (P < 0.05). The mean plasma [alanine] before exercise remained similar during all CDP (P > 0.05). Mean plasma [alanine] of the horses was increased after all exercise sessions measured (P < 0.05). There was no difference among conditioning programs (P > 0.05). It is concluded that conditioning with the exercise types used had small effects. This could have been because the exercise stress was too small, but also because the workload was not increased during the CDP. PMID- 19542498 TI - Direct and maternal genetic effects on first litter size, maturation age, and animal size in Finnish minks. AB - Variance components were estimated for maturing age, first litter size, and animal size in Finnish minks. The fitted animal models had direct genetic and maternal genetic effects, litter effects, and maternal environmental effects. Multivariate analysis was performed to determine covariances between the traits. Maternal effects represented a significant source of phenotypic variance in the maturation age and animal size. For litter size, maternal effects were not as clear. Moreover, in maturation age and animal size, the covariance between the direct additive effect and the maternal additive effect was negative. In addition, litter effect variances were larger than maternal variances for all traits. Therefore, it is crucial to also estimate environmental effects common to littermates for these traits. Direct heritability and the response to selection are overestimated, especially for maturation age and also for animal size, when maternal and common litter effects are not considered. PMID- 19542499 TI - Effect of rearing system and of dietary protein level on leptin, growth, and carcass composition in young Podolian bulls. AB - The present study aimed to evaluate the effects of rearing system and level of dietary protein supplementation on growth performance, metabolic profile, plasma leptin, and carcass composition of young Podolian bulls. At the beginning of the finishing period (about 14 mo of age), animals were divided into 3 groups according to rearing system and dietary protein level: indoor receiving a diet with 15% CP of DM (IND); and outdoor, at pasture, receiving a supplementation with 12% CP of DM (OUT12) or with 15% CP of DM (OUT15). Plasma leptin concentration increased (P < 0.05) with age, whereas it was unaffected by rearing system and protein level. Positive correlations between plasma leptin concentration, ADG (r = 0.861, P < 0.001), and blood glucose concentration (r = 0.977, P < 0.001) were observed, whereas NEFA and triglycerides were not correlated with leptin concentration. At the end of the finishing period (about 20 mo of age), young bulls in the OUT12 group showed greater (P < 0.05) cholesterol concentrations in blood than the OUT15 group, whereas the OUT15 group showed greater urea concentrations compared with IND (P < 0.01) and OUT12 groups (P < 0.001). Average daily gains, feed conversion ratio, and final BW were similar between groups. Intramuscular lipid content of LM, semitendinosus, and semimembranosus muscles was correlated with plasma leptin before slaughtering (r = 0.77, P < 0.01; r = 0.67, P < 0.01, r = 0.79, respectively). Correlations between plasma leptin and intramuscular fat were found starting from 18 mo of age for LM (r = 0.44; P < 0.05) and from 19 mo for semimembranosus (r = 0.56; P < 0.05) and semitendinosus (r = 0.58; P < 0.05). Mean value of intramuscular fat and intramuscular fat of LM was greater in OUT15 than OUT12 (P < 0.05). No differences were found between 12 and 15% CP supplementations for growth and carcass data. Therefore, we conclude that the extensive rearing system supplemented with 12% CP may be adequate to sustain good growth performance and carcass quality from the Podolian breed. These data also provide confirmation that leptin, in cattle, is directly related to nutritional status and may represent a suitable predictor for body fat starting from 1 mo before slaughtering. PMID- 19542500 TI - Effect of sward dry matter digestibility on methane production, ruminal fermentation, and microbial populations of zero-grazed beef cattle. AB - Increasing the digestibility of pasture for grazing ruminants has been proposed as a low-cost practical means of reducing ruminant CH(4) emissions. At high feed intake levels, the proportion of energy lost as CH(4) decreases as the digestibility of the diet increases. Therefore, improving forage digestibility may improve productivity as DM and energy intake are increased. A zero-grazing experiment was conducted to determine the effect of sward DM digestibility (DMD) on DMI, CH(4) emissions, and indices of rumen fermentation of beef animals. Twelve Charolais-cross heifers were assigned to 1 of 2 treatments, with 6 heifers per dietary treatment. Additionally, 4 cannulated Aberdeen Angus-cross steers were randomly allocated to each of these 2 treatments in a crossover design. Dietary treatments consisted of swards managed to produce (i) high digestibility pasture (high DMD) or (ii) pasture with less digestibility (low DMD), both offered for ad libitum intake. All animals were zero-grazed and offered freshly cut herbage twice daily. In vitro DMD values for the high and low DMD swards were 816 and 706 g/kg of DM. Heifers offered the high DMD grass had greater (P < 0.001) daily DMI of 7.66 kg compared with 5.38 kg for those offered the low DMD grass. Heifers offered the high DMD grass had greater (P = 0.003) daily CH(4) production (193 g of CH(4)/d) than those offered the low DMD grass (138 g of CH(4)/d). However, when corrected for DMI, digestible DMI, or ingested gross energy, there was no difference (P > 0.05) in CH(4) production between dietary treatments. For cannulated steers, intake tended (P = 0.06) to be greater for the high DMD grass (5.56 vs. 4.27 kg of DM/d), but rumen protozoa (4.95 x 10(4)/mL; P = 0.62); rumen ammonia (34 mg of N/L; P = 0.24); rumen total VFA (103 mM; P = 0.58), and rumen pH (6.8; P = 0.43) did not differ between treatments. There was no difference in total bacteria numbers, relative expression of the mcrA gene, and numbers of cycles to threshold for fungi when determined using quantitative PCR between dietary treatments with mean values of 73.0 ng/microL, 0.958, and 21.75 C(T), respectively. Results of this study demonstrate that there was no difference in CH(4) production when corrected for intake or rumen fermentation variables of beef cattle offered a high or low digestibility sward. PMID- 19542501 TI - Effects of syndyphalin-33 on feed intake and circulating measures of growth hormone, cortisol, and immune cell populations in the recently weaned pig. AB - The synthetic met-enkephalin syndyphalin-33 (SD-33) increases feed intake in sheep and transiently increases circulating GH concentrations in sheep, rats, and pigs. Two experiments were performed to evaluate the effects of SD-33 on recently weaned pigs. In a preliminary experiment, pigs were administered SD-33 (0.5 micromol/kg, given intramuscularly) or saline immediately before a 3-h transport and subsequent placement into group pens. Treatment with SD-33 increased (P = 0.01) daily feed intake; cumulatively, pen intake over 7 d postweaning tended (P = 0.06) to be greater than in control pens. In Exp. 2, pigs were weaned and fitted with jugular catheters. The following day, pigs were treated with SD-33 or saline as described above. Transient increases (P < 0.05) in circulating concentrations of GH (at 1 and 1.5 h postinjection) and cortisol (at 3.5 and 4 h postinjection) were observed in pigs treated with SD-33 relative to controls. No difference in feed intake was observed between treatments over 4 d postinjection. Increased (P < 0.05) numbers of circulating neutrophils, lymphocytes, and monocytes were observed in both treatment groups over 4 d postinjection, and treatment with SD-33 tended (P = 0.07) to selectively increase monocyte numbers. Although SD-33 has potential to be used to increase feed intake and decrease the negative effects of stress during weaning in pigs, further investigation is needed to better understand the timing of effect and to rule out possible immunosuppressive effects. PMID- 19542502 TI - Effects of type of canola protein supplement on ruminal fermentation and nutrient flow to the duodenum in beef heifers. AB - Ruminal fermentation, nutrient digestion, and flows to the duodenum in growing cattle fed differently produced canola protein supplements were studied in a 4 x 4 Latin square design using Speckle Park heifers (initial BW = 451 +/- 26 kg). Canola protein supplement treatments consisted of the following: 1) 8.78% regular canola meal (RCM); 2) 9.25% RCM plus 1.80% canola oil (RCMO); 3) 11.1% canola presscake from biodiesel oil extraction (CPC); and 4) 8.14% high ruminally undegradable protein (RUP) canola meal (RUCM) plus 1.32% canola oil (RUCMO). Experimental diets also contained 39.9, 40.2, 39.9, and 39.9% barley grain; 31.7, 31.4, 31.2, and 31.4% barley silage; and 17.5, 15.2, 15.6, and 16.5% oat hulls for the RCM, RCMO, CPC, and RUCMO diets, respectively. Feeding the CPC, RCMO, and RUCMO diets decreased (P < or = 0.05) ruminal NH(3)-N concentration compared with feeding the RCM diet. Compared with the RCM diet, adding canola oil in the RCMO diet or residual oil in the CPC diet resulted in greater ruminal concentrations of propionate (P < or = 0.09). Additionally, feeding the RCMO diet also resulted in greater ruminal concentrations of acetate (P = 0.07), valerate (P = 0.06), and total VFA (P = 0.07) compared with the RCM diet. Also, compared with the RCM diet, heifers on the RUCMO diet had decreased acetate (P = 0.02) concentrations. The changes in ruminal concentrations of acetate and propionate resulted in reduced acetate:propionate ratios in the RCMO (P = 0.08), CPC (P = 0.02), and RUCMO (P < 0.01) diets. Ruminal digestion and flows of nutrients to the duodenum were not affected by dietary treatment. However, adding canola oil to the RCMO and RUCMO dietary treatments decreased the digestibility of ADF (P < or = 0.08) and NDF (P < or = 0.08) in the total tract compared with the RCM diet. Total tract digestibility of OM was also decreased (P = 0.02) in heifers fed the RUCMO compared with the RCM diet. Notwithstanding the different processing methods employed in making RCM, CPC, or RUCM, there were no differences among the diets for ruminally degraded protein, ruminal microbial protein synthesis, and the flow of N fractions to the duodenum. PMID- 19542503 TI - Interrelationships among growth, endocrine, immune, and temperament variables in neonatal Brahman calves. AB - Interrelationships among growth, endocrine, immune, and temperament variables were assessed in neonatal Brahman calves. The velocity upon exiting a working chute (exit velocity) of an animal was measured and used as an objective indicator of temperament to classify calves as calm, intermediate, or temperamental. Calves (n = 116) were weighed weekly between d 0 and 21 to 24, and blood samples were collected for plasma and serum on d 0, 1, 2, 7, 14, and 21 to 24 after birth to measure concentrations of immunoglobulins, cortisol, and epinephrine (EPI). Body weight increased from d 0 through d 21 to 24 (P < 0.001) with bulls (n = 60) having greater BW than heifers (n = 56; P = 0.02). Serum concentrations of cortisol were greatest on d 0 before declining (P < 0.001) over the ensuing 21 to 24 d and were not related to temperament (P = 0.89) or sex (P = 0.97). Concentrations of EPI were affected by time, with an increase in EPI concentrations in temperamental bulls between 2 and 14 d of age (P < 0.008). Concentrations of EPI were not affected by temperament (P = 0.44) or sex (P = 0.68). Serum immunoglobulin concentrations peaked on d 1 before declining (P < 0.01) but were not related to temperament (P = 0.40 to 0.68). Of the stress hormones measured (cortisol and EPI), only cortisol was associated with the early performance of the calf. Calf BW at d 21 to 24 and BW gain were positively associated with serum immunoglobulin concentrations, yet negatively associated with concentrations of cortisol. Serum immunoglobulin concentrations were negatively correlated with cortisol concentrations (r = -0.28; P = 0.003), yet positively associated with EPI concentrations (r = 0.51; P = 0.003). During the neonatal period in this study, there was no relationship of temperament with passive immunity or stress hormone concentrations; however, growth was positively associated with passive immunity and negatively associated with stress hormones. Measuring exit velocity as early in life as d 21 to 24 fails to accurately predict temperament at weaning in over 40% of Brahman calves. Our conclusion is that measurement of exit velocity should be done nearer to the time of weaning than to birth. These data can be beneficial in developing best management practices for young calves. PMID- 19542504 TI - Differences in mitochondrial efficiency between lines of mice divergently selected for heat loss. AB - Divergent selection for heat loss was applied to lines of mice for 15 generations (G) in 3 replicates. Selection resumed at G42 and continued through G51 across all replicates. At the end of G51, differences in heat loss and feed intake per unit of BW were approximately 56 and 34%, respectively, between high heat loss (MH) and low heat loss (ML) lines, as a percentage of the control line (MC) mean. Rates of liver mitochondrial respiration states, degree of coupling, and mitochondrial efficiency were measured in G58 using a Clark-type oxygen electrode to investigate possible causes of underlying variation in maintenance requirements. Body composition, BW, liver weight, feed intake, and residual feed intake (RFI) were also measured or calculated. Results reported here represent data from 197 mature male mice from all replicates. There were no differences in BW (P = 0.91) between the selection lines. Selection had an effect on lean percentage (P = 0.02), with MH mice being leaner. Fat percentage differences between the selection lines tended toward significance (P = 0.13). Livers of MH mice were approximately 13% larger than livers of ML mice (P = 0.01). An effect of selection was observed (P < 0.01) in feed intake per unit BW, with MH mice consuming 29% more feed than ML mice in G58. Differences in state 2 and state 4 respiration rates were significant (P = 0.01), whereas state 3 rates approached significance (P = 0.06). Mitochondria of MH mice respired at a greater rate than mitochondria of ML mice in all states of respiration; ML mice had respiratory control ratios that were, on average, 8% greater than MH mice (P = 0.14). Although this difference only tended toward significance, we suspect a greater degree of coupling of mitochondrial processes exists in ML animals. Mice selected for reduced heat loss had ADP:oxygen ratios that were approximately 20% greater than MH mice (P = 0.03). Therefore, greater mitochondrial efficiency was expressed in the ML animals. Within a line-replicate, there was no correlation between ADP:O and feed intake per unit BW (P = 0.71). In addition, no correlation of ADP:O and RFI existed (P = 0.92). Although the selection lines differed in mitochondrial traits, including overall mitochondrial efficiency (ADP:oxygen), these differences were not a significant underlying cause of variation in feed intake per unit BW or in RFI estimates. PMID- 19542505 TI - Effects of phenyllactic acid on growth performance, nutrient digestibility, microbial shedding, and blood profile in pigs. AB - Ninety weanling pigs in Exp. 1 (6.27 +/- 0.73 kg; 21 d of age) and 96 growing pigs in Exp. 2 (21.73 kg +/- 1.29 kg; 56 d of age) were used in two 42-d experiments to evaluate the effect of phenyllactic acid (PLA) on growth performance, apparent total tract digestibility (ATTD) of DM and N, fecal pH value, microbial shedding, and blood profiles. In Exp. 1, the 3 dietary treatments were 1) negative control (NC), 2) positive control (PC), NC + antibiotics, and 3) PLA, NC + 0.5% PLA. In Exp. 2, dietary treatments were 1) control diet (CON), 2) PLA-0.1, CON + 0.1% PLA, 3) PLA-0.2, CON + 0.2% PLA, and 4) PLA-0.3, CON + 0.3% PLA. In Exp. 1, pigs fed the PC and PLA diets had greater ADFI during the overall period (P < 0.05) and tended to have greater ADG and G:F from d 7 to 21 (P < 0.10) than those fed the NC diet. The ATTD of DM was greatest in pigs fed the PLA diet on d 20 and 41, and N digestibility on d 20 was greater in pigs fed the PLA diet (P < 0.05) than those fed the NC diet. The numbers of white blood cell and lymphocyte concentrations on d 42 were increased (P < 0.05) by the inclusion of antibiotics and PLA in the diet. In Exp. 2, G:F tended to increase when PLA was added (quadratic, P < 0.10). The ATTD of DM did not differ among treatments, but there was a tendency (quadratic, P < 0.10) for N digestibility to increase as PLA levels increased. The lymphocyte percentage on d 42 increased linearly as dietary PLA increased (P < 0.05). Additionally, the white blood cell counts on d 42 tended to increase as PLA levels increased (P < 0.10). In both experiments, there was no effect of treatment on the fecal pH or presence of Lactobacillus, but the number of Escherichia coli in feces on d 41 decreased in response to the addition of PLA [P < 0.05 and 0.001 (linear) in Exp. 1 and 2, respectively]. In conclusion, PLA can decrease the number of E. coli, and this novel dietary acid may have potential to stimulate the immune system for both weanling and growing pigs. Thus, it could be a good candidate as an alternative to antibiotics in pig diets. PMID- 19542506 TI - A comparison of consumer sensory acceptance, purchase intention, and willingness to pay for high quality United States and Spanish beef under different information scenarios. AB - Tests were performed to identify variation across consumer evaluation ratings for 2 types of beef (Spanish yearling bull beef and US Choice and Prime beef), using 3 information levels (blind scores; muscle fat content + production conditions; and all production data including geographical origin) and 3 consumer evaluation ratings (hedonic rating, willingness to pay, and purchase intention). Further testing was carried out to assess the extent to which expert evaluations converged with those of untrained consumers. Taste panel tests involving 290 consumers were conducted in Navarra, a region in northern Spain. The beef samples were 20 loins of Pyrenean breed yearling bulls that had been born and raised on private farms located in this Spanish region and 20 strip loins from high quality US beef that ranged from high Choice to average Prime US quality grades. The Spanish beef were slaughtered at 507 +/- 51 kg of BW and 366 +/- 23 d of age. The US beef proved more acceptable to consumers and received greater ratings from the trained panel, with greater scores for juiciness (3.33), tenderness (3.33), flavor (3.46), and fat content (5.83) than for Spanish beef (2.77, 2.70, 3.14, 1.17). The differences in sensory variable rating were more pronounced for the Spanish beef than for the US beef, always increasing with the level of information. The variation in the ratings across different information levels was statistically significant in the case of the Spanish beef, whereas the variation observed in the ratings of the US beef was highly significant in the willingness of consumers to pay a premium. Consumers who appreciated greater quality were also more willing to pay for the additional level of quality. PMID- 19542507 TI - Leucine/glutamic acid/lysine protein 1 is localized to subsets of myonuclei in bovine muscle fibers and satellite cells. AB - Skeletal muscle growth is accomplished chiefly through the actions of satellite cells, a heterogeneous population that includes the adult muscle stem cell. Located adjacent to a mature muscle fiber, satellite cells typically reside in a quiescent state. Little information exists detailing satellite cell regulation of reversible G(0). One member of the mitosin family of centromere proteins, LEK1 (leucine/glutamic acid/lysine protein 1), is present in the nucleus of nondividing mouse satellite cells. The objective of this study was to evaluate LEK1 as a marker of quiescent bovine satellite cells (BSC) in vitro and in vivo. The BSC were isolated from young bull calves (< or =7 d) and cultured in vitro for up to 9 d before fixation and immunostaining for LEK1. Results demonstrated that all myogenic cells contain the protein, with immunostaining primarily within the nucleus and immediate perinuclear region. Immunocytochemical detection of LEK1 in cryosections of mature cows revealed that the protein was present in a fraction of satellite cells and muscle fiber nuclei. Approximately 20% of Pax7 expressing satellite cells contained LEK1. An equivalent percentage of myonuclei, as defined by nuclei within a dystrophin boundary, contained nuclear LEK1. To gain insight into the functional role of LEK1, BSC were transiently transfected with plasmids coding for putative dominant inhibitory LEK1 proteins [DeltaLEK1(991) and DeltaLEK1(911)] and evaluated for cell proliferation. Both forms of DeltaLEK1 inhibited (P < 0.05) BSC proliferation, as indicated by a decrease in Ki67 immunopositive cells. In C2C12 myoblasts, DeltaLEK1(911) inhibited (P < 0.05) myoblast determination protein 1 (MyoD)-directed muscle gene transcriptional activity; DeltaLEK1(991) had no effect on TnI-Luc transcription. By contrast, both DeltaLEK1 fusion proteins inhibited myogenin expression in BSC without disrupting myoblast fusion. These results provide evidence that LEK1 serves to coordinate proliferation and differentiation in myogenic cells. Coupling the immunostaining pattern and functional data, we propose that LEK1 may serve as a useful marker for satellite cells that are preparing to fuse into adjacent fibers as well as an indicator of recently added myonuclei. PMID- 19542508 TI - Effect of corn processing method and corn wet distillers grains plus solubles inclusion level in finishing steers. AB - Two experiments were conducted to determine the effect of corn processing method and corn wet distillers grains plus solubles (WDGS) level on steer performance and metabolism. In Exp. 1, 480 crossbred steer calves (314 +/- 18 kg of BW) were used in a finishing experiment with a randomized complete block design and a 3 x 4 treatment structure. Diets were based on dry-rolled (DRC), high-moisture (HMC), or steam-flaked corn (SFC) with increasing levels of WDGS (0, 15, 27.5, or 40%; DM basis). A corn processing x WDGS level interaction (P < 0.01) was observed for ADG and G:F. Average daily gain and G:F increased linearly (P < 0.01) in steers fed DRC; ADG increased quadratically (P = 0.04) and G:F increased linearly (P = 0.02) in steers fed HMC; and ADG decreased quadratically (P = 0.02) with no change in G:F (P = 0.52) in steers fed SFC as WDGS increased. In Exp. 2, 7 ruminally fistulated steers (440 +/- 41 kg of BW) were used in a 6-period crossover design with 3 x 2 factorial treatment structure. Diets were the same as those fed in Exp. 1, except they contained only 2 levels of WDGS (0 or 40% of diet DM). Total tract starch digestibility was greater (P < 0.01) for steers fed SFC than for steers fed DRC or HMC. Minimum ruminal pH was less (P < 0.01) for steers fed SFC than for steers fed HMC or DRC. Variance of ruminal pH was different among all 3 processing methods with DRC < HMC < SFC (P < 0.10). In situ 22-h DM digestibility of DRC and HMC and starch digestibility of DRC were greater (P < 0.10) in steers fed DRC compared with steers fed HMC or SFC. Steers fed 0% WDGS had less (P < or = 0.02) intake of DM, OM, NDF, and ether extract compared with steers fed 40% WDGS. Total tract digestibility of DM and OM was greater (P < or = 0.08) and digestibility of ether extract tended (P = 0.11) to be less for steers fed 0% WDGS compared with steers fed 40% WDGS. Maximum ruminal pH and pH variance were greater (P < or = 0.08) in steers fed 0% WDGS. A corn processing x WDGS level interaction (P = 0.09) was observed for ruminal acetate to propionate ratio (A:P). Within diets containing 0% WDGS, A:P in steers fed SFC was less (P < or = 0.08). In diets containing 40% WDGS, A:P was similar between processing methods and not different from the SFC with 0% WDGS. The corn processing x WDGS level interaction observed in the finishing experiment may be due to the decreased ruminal A:P in DRC and HMC diets with 40% WDGS. PMID- 19542509 TI - Effects of dietary changes and yeast culture (Saccharomyces cerevisiae) on rumen microbial fermentation of Holstein heifers. AB - The effects of a dietary challenge to induce digestive upsets and supplementation with yeast culture on rumen microbial fermentation were studied using 12 Holstein heifers (277 +/- 28 kg of BW) fitted with a ruminal cannula, in a crossover design with 2 periods of 5 wk. In each period, after 3 wk of adaptation to a 100% forage diet, the dietary challenge consisted of increasing the amount of grain at a rate of 2.5 kg/d (as-fed basis) over a period of 4 d, until a 10:90 forage:concentrate diet was reached, and then it was maintained for 10 d. Between periods, animals were fed again the 100% forage diet without any treatment for 1 wk as a wash-out period. Treatments started the first day of each period, and they were a control diet (CL) or the same diet with addition of yeast culture (YC, Diamond V XPCLS). Digestive upsets were determined by visual observation of bloat or by a reduction in feed intake (as-fed basis) of 50% or more compared with intake on the previous day. Feed intake was determined daily at 24-h intervals during the adaptation period and daily at 2, 6, and 12 h postfeeding during the dietary challenge. Ruminal liquid samples were collected daily during the dietary challenge to determine ruminal pH at 0, 3, 6, and 12 h postfeeding, and total and individual VFA, lactic acid, ammonia-N, and rumen fluid viscosity at 0 and 6 h postfeeding. The 16s rRNA gene copies of Streptococcus bovis and Megasphaera elsdenii were determined by quantitative PCR. Foam height and strength of the rumen fluid were also determined the day after the digestive upset to evaluate potential foam production. A total of 20 cases (83.3%) of digestive upsets were recorded in both periods during the dietary challenge, all diagnosed due to a reduction in feed intake. Rumen fermentation profile at 0 h on the digestive upset day was characterized by low ruminal pH, which remained under 6.0 for 18 h, accompanied by elevated total VFA concentration and, in some cases, by elevated lactate concentration. Addition of YC during the dietary challenge did not affect the incidence (10 cases per treatment) or time (7.00 +/- 0.62 d) to digestive upset. However, YC reduced (P < 0.05) the foam strength on the day after digestive upset, suggesting potential benefits of reducing the risk of developing bloat. The proposed dietary challenge model was successful in causing a digestive upset as indicated by reduced feed intake, but the YC addition had no significant impact on rumen fermentation. PMID- 19542510 TI - Associations between portion size acceptability of beef cuts and ribeye area of beef carcasses. AB - Carcasses that do not conform to mainstream specifications (i.e., those with nonconforming ribeye area) may not achieve their full potential value. Research was conducted to evaluate the relationship between beef carcass LM area at the 12th and 13th rib interface (LMA) and portion size acceptability of other muscles in the carcass. Sixty beef carcass sides of varying LMA sizes (between 67.74 and 116.13 cm(2)) were fabricated to generate 14 individual muscle cuts (triceps brachii long head, infraspinatus, chuckeye complexus, pectoralis profundus, longissimus thoracis, latissimus dorsi, gluteus medius, longissimus lumborum, tensor fasciae latae, psoas major, semimembranosus, biceps femoris, semitendinosus, and vastus lateralis). Retail portion size (g/1.27-cm-thick steak) as well as face surface area and dimensions were recorded for each steak cut perpendicular at the midpoint of the longitudinal axis of each muscle. Subsequently, a nationwide survey was conducted with foodservice chefs and retail meat merchandisers to evaluate acceptability of portion sizes and dimensions of individual muscle cuts. Simple linear regression and nonparametric regression analyses were used to evaluate results of the carcass muscle evaluation and survey, respectively. Results demonstrated that LMA did not affect (P < 0.05) retail portion size of 7 of the 14 muscles (chuckeye complexus, pectoralis profundus, psoas major, semimembranosus, tensor fasciae latae, triceps brachii, and vastus lateralis). Similarly, LMA did not affect (P < 0.05) surface area of steak cross-sectional face areas from 7 of the 14 muscles (chuckeye complexus, psoas major, semimembranosus, tensor fasciae latae, infraspinatus, vastus lateralis, and latissimus dorsi). Muscles for which carcass LMA (P < 0.05) was related to portion size or surface area of portion steaks, or both, were included in the survey. Results of the survey demonstrated that portion size for many muscles were still acceptable to retail merchandisers and foodservice chefs, even though carcass LMA was outside the range of commercially acceptable sizes. Results of this study demonstrated that carcass LMA is not an accurate determinant of the size, and subsequent acceptability, of many other muscles of beef in the carcasses, and may not be a good determinant of value of the beef carcass. PMID- 19542511 TI - Supplemental energy and extruded-expelled cottonseed meal as a supplemental protein source for beef cows consuming low-quality forage. AB - Three experiments were conducted to evaluate the efficacy of supplemental energy and extruded-expelled cottonseed meal (ECSM; 30.6% CP; 44% NDF, 10.2% fat; DM basis) as a protein supplement (SUP) to spring-calving beef cows (n = 96; 535 kg initial BW; 5.4 initial BCS) consuming low-quality forage during late gestation and early lactation. Supplementation of ECSM was compared with 2 traditional cottonseed meal-based SUP. For all experiments, SUP provided equal CP. On a DM basis, SUP included 1) a blend of 76% wheat middlings and 18% solvent-extracted cottonseed meal (WMCSM); 2) solvent-extracted cottonseed meal (CSM); and 3) delinted, extruded-expelled cottonseed meal (ECSM). In Exp. 1, cows were individually fed SUP 3 d/wk until calving and 4 d/wk during lactation; total SUP period was 95 d. Tall-grass prairie hay (4.4% CP; 74% NDF; DM basis) was provided during the SUP period. Changes in cow BW during gestation (P = 0.23), over the SUP period (P = 0.27), and over the 301-d experiment (P = 0.56) were similar. Changes in BCS were similar during gestation (P = 0.78), over the SUP period (P = 0.95) and over the 301-d experiment (P = 0.37). Calf birth weights (P = 0.21) and BW at weaning (P = 0.76) were not different. Percentage of cows exhibiting luteal activity at the beginning of breeding season (P = 0.59), AI conception rate (P = 0.71), and pregnancy rate at weaning (P = 0.88) were not different. In Exp. 2, 18 cows in early lactation from Exp. 1 were used to determine the effect of SUP on hay intake and digestion. Hay intake tended (P = 0.10) to be greater for CSM than ECSM. Intake of OM and DM was greater for WMCSM (P 0.10). Similarly, 24-h milk production was not different (P = 0.25). Neither greater energy intake of cows consuming WMCSM nor greater fat intake of cows consuming ECSM influenced cow performance measures or calf weaning weight. Cow response to SUP with ECSM compared with traditional cottonseed meal-based SUP indicates that ECSM is a viable source of supplemental protein for beef cows consuming low quality forage. PMID- 19542512 TI - In situ ruminal degradation characteristics of by-product feedstuffs for beef cattle consuming low-quality forage. AB - Eight ruminally cannulated steers (BW = 753 +/- 48 kg) were used to evaluate in situ N, NDF, and DM degradation characteristics of by-product feeds and their application for beef cows consuming low-quality forage. Experimental feedstuffs included (DM basis) 1) extruded-expelled cottonseed meal (ECSM; 33% CP and 55% NDF), 2) extruded-expelled cottonseed meal with linters (ECSML; 25% CP and 41% NDF), 3) dried distillers grains with solubles (DGS; 33% CP and 36% NDF), 4) solvent-extracted cottonseed meal (CSM; 43% CP and 29% NDF), and 5) a blend of 76% wheat middlings with 18% CSM (WMCSM; 23% CP and 40% NDF). Steers were fed chopped prairie hay (4.8% CP, 69% NDF; DM basis) ad libitum and received 0.38 kg/100 kg of BW of WMCSM daily. In situ degradation kinetics of N, NDF, and DM components included the following fractions: A (immediately soluble), B (potentially degradable), and C (undegradable). Calculated rumen degradable protein (RDP) for ECSM was the greatest among all feedstuffs (83.8%; P < 0.01), which was composed of a large A fraction of N (41%). Similar RDP values were observed for DGS and ECSML (50.7 and 50.9%, respectively, P = 0.93). The B fraction N for ECSML was large (88.9%); however, most of this was unavailable for ruminal degradation. The amount of RDP in CSM and WMCSM was similar (78.2 and 73.5%, respectively; P = 0.12) though the A fraction of N was greater for WMCSM compared with CSM (P < 0.01). Degradability of NDF was greatest (P < 0.01) for DGS (67.4%) and was similar (P = 0.48) for WMCSM and CSM (54.5 and 57.0%, respectively). The least degradability of NDF was calculated for ECSM (29.3%; P < 0.01), attributed to greater lignin content (13.3%, DM). Degradability of DM was greatest (P < 0.01) for CSM and WMCSM (63.7 and 59.4%, respectively) and least (P < 0.01) for ECSM (36.5%) and ECSML (40.6%). Ruminal N degradation characteristics of ECSM were similar to more traditional supplements containing CSM and WMCSM. The RDP for ECSML and DGS N was less compared with other feedstuffs, indicating these feeds may need to be blended with other ingredients containing greater concentrations of degradable N, particularly in situations in which forage RDP is low. PMID- 19542513 TI - Metabolic maturity at birth and neonate lamb survival and growth: the effects of maternal low-dose dexamethasone treatment. AB - Perinatal mortality is a major contributing factor to reproductive wastage in grazing sheep industries. Enhanced metabolic and endocrine maturity at birth may improve the behavioral competency and thermoregulatory ability of neonates, potentially improving lamb survival over the first 72 h of life. Maternal glucocorticoid treatment in late gestation was investigated as a mechanism for manipulating metabolic and endocrine maturity in the ovine neonate. Multiparous, fine-wool Merino ewes (n = 150) were divided into 3 groups to lamb on pasture. Within each group, 5 single-lamb and 5 twin-lamb bearing ewes were randomly allocated to 1 of 5 treatments. Treatments included a saline control (1 mL), or dexamethasone (2 mg/mL as the sodium phosphate) injected intramuscularly at 1 of 2 dose rates (1.5 or 3.0 mg) at d 130 or 141 of gestation. One-half of the control ewes were injected at d 130 and the remainder at d 141. Dexamethasone treatment had no effect on lamb survival to 72 h after birth, although there tended (P = 0.09) to be a smaller proportion of lambs dying due to dystocia than for control lambs. Heart girth at birth in singleton and twin lambs was reduced (P < 0.01) at the greater dose rate. Further, treatment also reduced birth weight (by about 5%) and presuckling rectal temperatures in twin lambs, but not in singleton lambs. These reductions were also dependent on the sex of the lamb. Dexamethasone treatment did not alter gestation length or lamb presuckling plasma glucose, NEFA, urea, or leptin concentrations, but treatment at d 141 increased (P < 0.05) ghrelin concentrations in singleton and male lambs. Behavioral interactions between ewes and neonatal lambs were generally unaffected, although treatment at d 130 produced lambs that took longer to bleat than lambs of untreated ewes (P < 0.05). Treatment did not affect the concentration of measured blood metabolites or hormones at weaning. Although there were interactions between litter size, lamb sex, and the dose rate and time of treatment on weaning weight, BW recorded 73 d after weaning was unaffected by treatment. Despite changes in birth weight, rectal temperature, lamb behavior, and presuckling plasma ghrelin concentrations, survival in the first 72 h of life, and lamb growth performance were unaffected by periparturient maternal glucocorticoid treatment. PMID- 19542514 TI - Genetic relationships among pig behavior, growth, backfat, and loin muscle area. AB - The objective of this study was to estimate repeatabilities and heritabilities for measures of pig behavior and their relationship with performance. Measures of behavior and performance included the backtest, resident-intruder test, human approach test (HAT), novel object test (NOT), d 1 BW, backfat depth (BF), loin muscle area (LMA), ADG in the farrowing house, ADG, 21-d BW, and 140-d BW (W). Each behavioral trait was measured twice. The study consisted of 95 litters from 31 sires with an average of 3 litters per sire (n >or= 457). Between 7 and 14 d of age, the backtest was conducted by placing each pig in a supine position for 60 s. Total time spent struggling (TTS) and total number of attempts to struggle (TAS) were recorded. The resident intruder test involved 2 nursery pigs, a resident pig and an unfamiliar intruder pig. The resident pen was divided in half by a solid partition. A resident pig was placed in the test area, and an intruder pig was then introduced. Latency until an attack occurred (LAT) and total number of attacks over 2 tests (RIS) were recorded. Amount of time taken for each finishing pig to make snout contact with an unfamiliar human or object was recorded. Dam and sire effects influenced all traits (P < 0.01). Sex and pen affected LAT, RIS, HAT, and NOT (P < 0.10). Repeatabilities of TTS, TAS, RIS, LAT, HAT, and NOT were 0.38, 0.21, 0.07, 0.08, 0.17, and 0.11, respectively. The phenotypic correlations of TTS with TAS and HAT with NOT were 0.61 and 0.34, respectively. Phenotypic correlation between RIS and LAT was -0.85. Total time spent struggling and TAS tended to be phenotypically correlated with 21-d BW and ADG in the farrowing house. Total attempts to struggle were phenotypically correlated with BF (0.15). Latency until an attack occurred was phenotypically correlated with LMA (0.23). Resident intruder score was phenotypically correlated with ADG (-0.13), W (-0.13), and LMA (-0.21) and estimated to be lowly heritable (h(2) = 0.12). Heritabilities of TTS and TAS were 0.31 and 0.53, respectively. Genetic correlation of TAS with ADG and W was 0.38. Genetic correlations of TTS with BF, W, and TAS were 0.14, 0.18, and 0.81, respectively. The backtest is a heritable and repeatable measure of a behavioral characteristic in pigs that is phenotypically and genetically correlated with performance. PMID- 19542515 TI - Bovine myofiber characteristics are influenced by postweaning nutrition. AB - This study determined the extent to which bovine longissimus lumborum muscle (LLM) myofibers are influenced by nutrition for 120 d from weaning and the time course of recovery after severe postweaning nutritional restriction. After weaning, 3 groups of Belmont Red cattle, a tropically adapted breed, were fed to achieve rapid growth (RG, > or =0.6 kg of BW gain/d; n = 16), slow growth (SG, 0.2 kg of BW gain/d; n = 17), or BW loss (WL, 10% loss of weaning weight; n = 17) over 120 d. They were then grazed as 1 group at pasture with forage supplementation for 600 d until slaughter at approximately 500 kg of BW. Samples of LLM were taken from 8 to 12 animals per treatment 6 d before (baseline) and 115, 204, 324, and 476 d after commencement of the study and from all cattle at slaughter (d 721). Myofiber characteristics were determined by immunocytochemical staining of myosin heavy chains. Cross-sectional areas (CSA) of the major myofiber types 1, 2A, and 2X in WL were reduced at d 115 compared with baseline and with the growth groups (all P < 0.001); however, there was little difference in the percentage of the different myofiber types (all P > 0.10). Differences in CSA of the major myofiber types between WL and the growth groups at 115 d were smallest for type 1 (slow oxidative) and greatest for type 2X (fast glycolytic). Consequently, the relative area (percentage of total myofiber area) of type 1 myofibers in WL was significantly greater at 115 d than in the growth groups (P < 0.001). During recovery from postweaning nutritional restriction, significant differences in major myofiber type percentages were not evident (all P > 0.10), and by 721 d CSA of myofiber types differed little between the treatment groups, although SG had greater CSA of type 1 (P < 0.05) and type 2A (P < 0.01) myofibers than WL and RG. At 721 d, the relative area of type 2A myofibers was less in WL compared with SG (P < 0.01) and RG (P < 0.05) and of type 2X myofibers greater (P < 0.05) in WL compared with SG. It is concluded that in the LLM of cattle undergoing severe nutritional restriction immediately postweaning, the size of the more glycolytic fiber types is more adversely affected than the more oxidative types, resulting in an increased relative area of type 1, slow oxidative myofibers. However, given adequate time and nutriment at pasture, LLM myofiber characteristics of cattle recovered to near normal after severe, chronic nutritional restriction immediately postweaning, consistent with earlier findings for beef quality. PMID- 19542516 TI - Movie smoking and urge to smoke among adult smokers. AB - INTRODUCTION: Few studies have assessed the association between exposure to movie smoking and urge to smoke under real-world conditions. METHODS: We conducted exit interviews with 4,073 movie patrons, of whom 2,817 were aged 18 years or older. Some 536 were smokers and had complete data. Subjects had exited 26 movies, of which 12 contained smoking. We used least squares regression to assess the association between exposure to movie smoking and urge to smoke (scale range 0 10), controlling for movie rating, age, sex, heaviness of smoking index (HSI, range 0-6), and time since last cigarette smoked. RESULTS: Median age was 27 years and 52% were female. Median urge to smoke level at movie exit was 7. The dose-response between higher categories of movie smoking and median urge to smoke was one point for two lower categories (1-11 and 11-54 s) and two for the highest category (>or=55 s), but these differences were not statistically significant. In the multivariate analysis, attendance of a movie with smoking was associated with a 0.81-point increase (95% CI = 0.46-1.16) in urge to smoke. For comparison, an HSI score of 3 (vs. 0) was associated with a 2-point increase in urge to smoke. DISCUSSION: In this sample of adult smokers, exposure to movie smoking was associated with higher urge to smoke after the movie, independent of movie rating. The effect size was consistent with responses seen in cue reactivity experiments. Exposure to movie smoking may affect urge to smoke among adult smokers. PMID- 19542517 TI - Continuous individual support of smoking cessation using text messaging: a pilot experimental study. AB - INTRODUCTION: The objective of this study was to test the feasibility and acceptance of an intervention using text messaging (short message service [SMS]) for continuous individual support of smoking cessation in young adults. Additionally, the optimal feedback intensity was investigated, and short-term efficacy of the intervention was explored. METHODS: In a cafeteria of the University of Greifswald, 575 visitors were screened for smoking status and usage of text messaging. From these, 194 persons who fulfilled the inclusion criteria of daily smoking and weekly usage of SMS were invited for participation in an SMS based intervention. From these, 174 (90%) consented to participate. The participants were randomly allocated to one of three study groups: (a) control condition without intervention, (b) intervention with one weekly SMS feedback (1SMS), or (c) intervention with three weekly SMS feedbacks (3SMS). In study groups (b) and (c), individualized SMS feedbacks were sent to the participants weekly, based on data from the baseline assessment and a weekly SMS assessment of the stages of change according to the transtheoretical model. Program use and acceptance were compared between the two intervention groups differing in support intensity. An exploration of the short-term efficacy of the program was conducted by comparing the three study groups at the end of the 3-month intervention program on smoking variables. RESULTS: The median number of replies to the weekly SMS assessments was 12.5 in the 1SMS group and 13.0 in the 3SMS group (not significant). The acceptance of the program did not differ between the intervention groups. At postassessment, no significant differences between the three study groups emerged on the examined smoking variables. DISCUSSION: The high participation and retention rates suggest that SMS-based smoking cessation interventions are attractive for young adults. Support intensity did not affect the acceptance of the program. Longer follow-up periods and larger samples are required to obtain conclusive results about the efficacy of this intervention approach. PMID- 19542518 TI - Psychometric properties of the Wisconsin Inventory of Smoking Dependence Motives (WISDM-68): a replication and extension. AB - Introduction The Wisconsin Inventory of Smoking Dependence Motives (WISDM-68), a relatively new measure, assesses nicotine dependence in terms of distinct motivations for smoking. We examined psychometric properties of the WISDM-68 in a population-based sample that is on average older and includes heavier smokers than the original sample used for the validation of the instrument. Methods Participants were adult regular smokers (N = 431) who were offspring of pregnant women enrolled in the New England sites of the National Collaborative Perinatal Project (1959-1966). We examined the internal consistency of the WISDM-68's 13 subscales, replicated and extended the confirmatory factor analysis (CFA) by Piper et al., assessed the interdependence of the subscales, examined the association between smoking heaviness and subscale scores, and conducted additional validation tests. Results Internal consistency for WISDM's 13 subscales ranged from 0.78 for the Tolerance to 0.89 for the Cognitive Enhancement and Affiliative Attachment subscales. Similar reliabilities were obtained for demographic and smoking-relevant subgroups. CFAs suggest that a 13 factor model fit our data better than a single-factor model and better than an empirically derived 10-factor model. Regression models supported the validity of the 13 subscales, although follow-up analyses suggested possibility of maintaining WISDM's 13-factor structure with fewer than 68 items. Conclusion The WISDM-68 consists of 13 internally consistent subscales. The independence of the majority of the subscales supports the perspective that nicotine dependence is a heterogeneous construct. PMID- 19542519 TI - Aging among Jewish Americans: implications for understanding religion, ethnicity, and service needs. AB - PURPOSE: This article challenges popular conceptions of the nature of ethnicity and religiousness in the gerontological literature. Using the example of older Jewish Americans, the authors argue for more nuanced definitions and usage of terms such as "religion" and "ethnicity" in order to begin to understand the complex interweaving of these two dimensions in the lives of older persons. DESIGN AND METHODS: The analyses used data from the 2000-2001 National Jewish Population Survey (NJPS) as well as comparisons with the 1990 NJPS. There were 1,099 respondents aged 65 years and older in the 2000-2001 NJPS who reported themselves to be Jewish. This sample was then split into three groups: those who reported observing only Judaism and were affiliated with a denomination within Judaism (n = 776) or were not affiliated (n = 277) and those who reported observing another faith in addition to Judaism (n = 46). RESULTS: Respondents to the 2000-2001 NJPS were older, wealthier, and less likely to be members of a religious denomination than those in the 1990 NJPS. Denominational affiliates were more likely than the other two groups to have a strong ethnic identity but less likely to indicate that religion was important in their lives. Denomination members were also more likely to be children or grandchildren of immigrants. IMPLICATIONS: Several key assumptions in the study of ethnicity and religiousness in much of current gerontological research need reassessment especially assumptions about the links between religious identification, beliefs, practices, and communal solidarity. PMID- 19542520 TI - Wisdom of generations: a pilot study of the values transmitted in ethical wills of nursing home residents and student volunteers. AB - PURPOSE: This is a pilot study that provides a description of the values older persons report in ethical wills and their reasoning for the values they chose, and compares the values in ethical wills of seniors and students. Nursing home residents rarely get the opportunity or venue to discuss these topics and the ethical will enables them to have conversations about issues they feel are important. DESIGN AND METHODS: The 22 Questions for Ethical Wills(c) assessment was administered to 15 residents of a large nursing home in suburban Maryland and to 11 student volunteers. Raters identified recurring themes and independently analyzed the text from the ethical wills based on the themes. Questions that prompted similar responses were combined in the analyses. RESULTS: The most prevalent theme among both nursing home residents and students was "interpersonal relations." Differences between groups of respondents were especially evident for the theme of "education," which was given by over a half of the older persons and none of the students. Greater variability was found in the responses of the older participants, as was a greater likelihood of providing detailed explanations for their relayed values. IMPLICATIONS: The 22 Questions for Ethical Wills(c) is a useful methodology to elicit meaningful discussions of values and life lessons in persons both young and old. This process offers an intriguing comparison between the similarities and differences of life views of persons at opposite ends of the age spectrum. PMID- 19542522 TI - A novel role for Gtb1p in glucose trimming of N-linked glycans. AB - Glucosidase II (GluII) is a glycan-trimming enzyme active on nascent glycoproteins in the endoplasmic reticulum (ER). It trims the middle and innermost glucose residues (Glc2 and Glc1) from N-linked glycans. The monoglucosylated glycan produced by the first GluII trimming reaction is recognized by calnexin/calreticulin and serves as the signal for entry into this folding pathway. GluII is a heterodimer of alpha and beta subunits corresponding to yeast Gls2p and Gtb1p, respectively. While Gls2p contains the glucosyl hydrolase active site, the Gtb1p subunit has previously been shown to be essential for the Glc1 trimming event. Here we demonstrate that Gtb1p also determines the rate of Glc2 trimming. In order to further dissect these activities we mutagenized a number of conserved residues across the protein. Our data demonstrate that both the MRH and G2B domains of Gtb1p contribute to the Glc2 trimming event but that the MRH domain is essential for Glc1 trimming. PMID- 19542523 TI - A simple micro-method for determining precise oligosaccharidic specificity of mannose-binding lectins. AB - A simple and inexpensive method was developed to rapidly define the specificity of mannose-specific lectins toward oligomannoside-type structures. The method involved the interaction of a mixture of N-[(14)C]-acetylated glycoasparagines, prepared by exhaustive pronase digestion of bovine pancreatic ribonuclease B and N-[(14)C]-acetylation with [(14)C]-acetic anhydride and containing all the possible oligomannoside-type N-glycans, with the lectin immobilized on Sepharose 4B. After exhaustive desalting, the obtained fractions were separated by high performance thin-layer chromatography on silica gel plates and visualized by autoradiography with intensifying screen. As an example of the usefulness of this method, the fine specificity of artocarpin, the mannose-specificity lectin isolated from seeds of jackfruit (Artocarpus integrifolia) toward oligomannoside type structures is presented. On the basis of such a determination, the best oligomannosidic ligand recognized by a mannose-specific lectin can be selected for studies of crystal structures of the lectin in complex with the defined ligand. Furthermore, some of these immobilized lectins, after definition of their precise specificities with the method, could represent valuable tools for the fractionation and characterization of oligomannose-type structures, present in complex mixtures. PMID- 19542524 TI - The specific localization of seminolipid molecular species on mouse testis during testicular maturation revealed by imaging mass spectrometry. AB - More than 90% of the glycolipid in mammalian testis consists of a unique sulfated glyceroglycolipid called seminolipid. The galactosylation of the molecule is catalyzed by UDP-galactose:ceramide galactosyltransferase (CGT). Disruption of the CGT gene in mice results in male infertility due to the arrest of spermatogenesis, indicating that seminolipid plays an important role in reproductive function. Seminolipid molecules can be assigned to different molecular species based on the fatty acid composition. In this report, we investigated the localizations of the molecular species of seminolipid by imaging mass spectrometry and demonstrated that major molecule (C16:0-alkyl-C16:0-acyl) was expressed throughout the tubules: some (C16:0-alkyl-C14:0-acyl and C14:0 alkyl-C16:0-acyl) were predominantly expressed in spermatocytes and the other (C17:0-alkyl-C16:0-acyl) was specifically expressed in spermatids and spermatozoa. This is the first report to show the cell-specific localization of each molecular species of seminolipid during testicular maturation. PMID- 19542525 TI - Ziprasidone vs olanzapine in recent-onset schizophrenia and schizoaffective disorder: results of an 8-week double-blind randomized controlled trial. AB - INTRODUCTION: Head-to-head comparisons of antipsychotics have predominantly included patients with chronic conditions. The aim of the present study was to compare the efficacy and tolerability of ziprasidone and olanzapine in patients with recent-onset schizophrenia. METHODS: The study was an 8-week, double-blind, parallel-group, randomized, controlled multicenter trial (NCT00145444). Seventy six patients with schizophreniform disorder, schizophrenia or schizoaffective disorder (diagnosis < 5 y), and a maximum lifetime antipsychotic treatment < 16 weeks participated in the study. Efficacy of ziprasidone (80-160 mg/d) and olanzapine 10-20 mg was measured using the Positive and Negative Syndrome Scale (PANSS), the Clinical Global Impression (CGI) Scale, the Calgary Depression Scale for Schizophrenia (CDSS), and the Heinrich Quality of Life Scale (HQLS); tolerability assessments included laboratory assessments, body weight, and electroencephalogram. RESULTS: Olanzapine (n = 34) and ziprasidone (n = 39) showed equal efficacy as measured by the PANSS, CDSS, CGI, and HQLS. However, mean weight gain was significantly higher in the olanzapine group (6.8 vs 0.1 kg, P < .001). Ziprasidone was associated with decreasing levels of triglycerides, cholesterol, and transaminases, while these parameters increased in the olanzapine group (all P values < .05). There were no significant differences in fasting glucose and prolactin levels or in cardiac or sexual side effects. Patients on ziprasidone used biperiden for extrapyramidal side effects more frequently (P < .05). DISCUSSION: The results of this study indicate that ziprasidone and olanzapine have comparable therapeutic efficacy but differ in their side effect profile. However, there is a risk of a type II error with this sample size. Clinically significant weight gain and laboratory abnormalities appear early after initiating treatment and are more prominent with olanzapine, while more patients on ziprasidone received anticholinergic drugs to treat extrapyramidal symptoms. PMID- 19542526 TI - Hippocampi, thalami, and accumbens microstructural damage in schizophrenia: a volumetry, diffusivity, and neuropsychological study. AB - Volumetric abnormalities in the subcortical structures have been described in schizophrenia. However, it still has to be clarified if subtle microstructural damage is also present. Thus, we aimed to detect subcortical volume and mean diffusivity (MD) alterations in 45 patients with diagnosis of schizophrenia compared with 45 age-, gender-, and educational attainment-matched healthy comparison (HC) participants, by using a combined volumetry and diffusion tensor imaging (DTI) method. A secondary aim was to identify the neuropsychological correlates of subcortical abnormalities in the schizophrenic group. We found thalami and hippocampi bilaterally and left accumbens to show MD increase in the schizophrenic group. No volumetric decrease was found. Moreover, significant correlations between the MD values in subcortical structures (right thalamus and hippocampus and left accumbens) and working memory performance were found. Thus, subcortical microstructural alterations are present in schizophrenia even in absence of volumetric abnormalities. Furthermore, microstructural damage in subcortical areas is linked to working memory, suggesting the presence of a subtle microstructural subcortical dysfunction in the pathoetiological mechanism underlying high cognitive load performances in schizophrenia. Finally, our findings indicate that MD is a more sensitive marker of brain tissue deficits than signal intensity variations measured in T1-weighted imaging data, consistently with previous reports. Thus, DTI appears to be an invaluable tool to investigate subcortical pathology in schizophrenia, greatly enhancing the ability to detect subtle brain changes in this complex disorder. PMID- 19542527 TI - Are screening instruments valid for psychotic-like experiences? A validation study of screening questions for psychotic-like experiences using in-depth clinical interview. AB - Individuals who report psychotic-like experiences are at increased risk of future clinical psychotic disorder. They constitute a unique "high-risk" group for studying the developmental trajectory to schizophrenia and related illnesses. Previous research has used screening instruments to identify this high-risk group, but the validity of these instruments has not yet been established. We administered a screening questionnaire with 7 items designed to assess psychotic like experiences to 334 adolescents aged 11-13 years. Detailed clinical interviews were subsequently carried out with a sample of these adolescents. We calculated sensitivity and specificity and positive predictive value (PPV) and negative predictive value (NPV) for each screening question for the specific symptom it enquired about and also in relation to any psychotic-like experience. The predictive power varied substantially between items, with the question on auditory hallucinations ("Have you ever heard voices or sounds that no one else can hear?") providing the best predictive power. For interview-verified auditory hallucinations specifically, this question had a PPV of 71.4% and an NPV of 90.4%. When assessed for its predictive power for any psychotic-like experience (including, but not limited to, auditory hallucinations), it provided a PPV of 100% and an NPV of 88.4%. Two further questions-relating to visual hallucinations and paranoid thoughts-also demonstrated good predictive power for psychotic-like experiences. Our results suggest that it may be possible to screen the general adolescent population for psychotic-like experiences with a high degree of accuracy using a short self-report questionnaire. PMID- 19542528 TI - Characterization of cis-9 trans-11 trans-15 C18:3 in milk fat by GC and covalent adduct chemical ionization tandem MS. AB - Rumen biohydrogenation of dietary alpha-linolenic acid gives rise in ruminants to accumulation of fatty acid intermediates, some of which may be transferred into milk. Rumelenic acid [cis-9 trans-11 cis-15 C18:3 (RLnA)] has recently been characterized, but other C18:3 minor isomers are still unknown. The objective of this work was to identify a new isomer of octatridecenoic acid present in milk fat from ewes fed different sources of alpha-linolenic acid. Structural characterization of this fatty acid was achieved by GC-MS. Analysis of dimethyloxazoline and picolinyl ester derivatives allowed for location of the double bond positions. Covalent adduct chemical ionization tandem mass spectrometry confirmed the positional structure 9-11-15, identical to RLnA, and helped to establish double bond geometry (cis-trans-trans). This new C18:3 isomer could be formed by isomerization of cis-15 bond of RLnA and subsequently converted by hydrogenation to trans-11 trans-15 C18:2, an octadecadienoic acid also detected in this study. PMID- 19542529 TI - The barrel cortex as a model to study dynamic neuroglial interaction. AB - There is increasing evidence that glial cells, in particular astrocytes, interact dynamically with neurons. The well-known anatomofunctional organization of neurons in the barrel cortex offers a suitable and promising model to study such neuroglial interaction. This review summarizes and discusses recent in vitro as well as in vivo works demonstrating that astrocytes receive, integrate, and respond to neuronal signals. In addition, they are active elements of brain metabolism and exhibit a certain degree of plasticity that affects neuronal activity. Altogether these findings indicate that the barrel cortex presents glial compartments overlapping and interacting with neuronal compartments and that these properties help define barrels as functional and independent units. Finally, this review outlines how the use of the barrel cortex as a model might in the future help to address important questions related to dynamic neuroglia interaction. PMID- 19542531 TI - The use of sedation to relieve cancer patients' suffering at the end of life: addressing critical issues. PMID- 19542530 TI - Secreted versus membrane-anchored collagenases: relative roles in fibroblast dependent collagenolysis and invasion. AB - Fibroblasts degrade type I collagen, the major extracellular protein found in mammals, during events ranging from bulk tissue resorption to invasion through the three-dimensional extracellular matrix. Current evidence suggests that type I collagenolysis is mediated by secreted as well as membrane-anchored members of the matrix metalloproteinase (MMP) gene family. However, the roles played by these multiple and possibly redundant, degradative systems during fibroblast mediated matrix remodeling is undefined. Herein, we use fibroblasts isolated from Mmp13(-/-), Mmp8(-/-), Mmp2(-/-), Mmp9(-/-), Mmp14(-/-) and Mmp16(-/-) mice to define the functional roles for secreted and membrane-anchored collagenases during collagen-resorptive versus collagen-invasive events. In the presence of a functional plasminogen activator-plasminogen axis, secreted collagenases arm cells with a redundant collagenolytic potential that allows fibroblasts harboring single deficiencies for either MMP-13, MMP-8, MMP-2, or MMP-9 to continue to degrade collagen comparably to wild-type fibroblasts. Likewise, Mmp14(-/-) or Mmp16(-/-) fibroblasts retain near-normal collagenolytic activity in the presence of plasminogen via the mobilization of secreted collagenases, but only Mmp14 (MT1 MMP) plays a required role in the collagenolytic processes that support fibroblast invasive activity. Furthermore, by artificially tethering a secreted collagenase to the surface of Mmp14(-/-) fibroblasts, we demonstrate that localized pericellular collagenolytic activity differentiates the collagen invasive phenotype from bulk collagen degradation. Hence, whereas secreted collagenases arm fibroblasts with potent matrix-resorptive activity, only MT1-MMP confers the focal collagenolytic activity necessary for supporting the tissue invasive phenotype. PMID- 19542532 TI - Palliative sedation therapy does not hasten death: results from a prospective multicenter study. AB - BACKGROUND: Palliative sedation therapy (PST) is indicated for and used to control refractory symptoms in cancer patients undergoing palliative care. We aimed to evaluate whether PST has a detrimental effect on survival in terminally ill patients. METHODS: This multicenter, observational, prospective, nonrandomized population-based study evaluated overall survival in two cohorts of hospice patients, one submitted to palliative sedation (A) and the other managed as per routine hospice practice (B). Cohorts were matched for age class, gender, reason for hospice admission, and Karnofsky performance status. RESULTS: Of the 518 patients enrolled, 267 formed cohort A and 251 cohort B. In total, 25.1% of patients admitted to the participating hospices received PST. Mean and median duration of sedation was 4 (standard deviation 6.0) and 2 days (range 0-43), respectively. Median survival of arm A was 12 days [90% confidence interval (CI) 10-14], while that of arm B was 9 days (90% CI 8-10) (log rank = 0.95, P = 0.330) (unadjusted hazard ratio = 0.92, 90% CI 0.80-1.06). CONCLUSION: PST does not shorten life when used to relieve refractory symptoms and does not need the doctrine of double effect to justify its use from an ethical point of view. PMID- 19542533 TI - Immune thrombocytopenic purpura (ITP) and breast cancer. Does adjuvant therapy for breast cancer improve platelet counts in ITP? PMID- 19542534 TI - Successful treatment of ifosfamide-induced hyponatremia with AVP receptor antagonist without interruption of hydration for prevention of hemorrhagic cystitis. PMID- 19542536 TI - BRCA2 splice site mutations in an Italian breast/ovarian cancer family. PMID- 19542537 TI - CK2-dependent phosphorylation determines cellular localization and stability of ataxin-3. AB - The nuclear presence of the expanded disease proteins is of critical importance for the pathogeneses of polyglutamine diseases. Here we show that protein casein kinase 2 (CK2)-dependent phosphorylation controls the nuclear localization, aggregation and stability of ataxin-3 (ATXN3), the disease protein in spinocerebellar ataxia type 3 (SCA3). Serine 340 and 352 within the third ubiquitin-interacting motif of ATXN3 were particularly important for nuclear localization of normal and expanded ATXN3 and mutation of these sites robustly reduced the formation of nuclear inclusions; a putative nuclear leader sequence was not required. ATXN3 associated with CK2alpha and pharmacological inhibition of CK2 decreased nuclear ATXN3 levels and the formation of nuclear inclusions. Moreover, we found that ATXN3 shifted to the nucleus upon thermal stress in a CK2 dependent manner, indicating a key role of CK2-mediated phosphorylation of ATXN3 in SCA3 pathophysiology. PMID- 19542538 TI - Intra-His bundle block in second-degree Mobitz I atrioventricular block with right bundle branch block. AB - Because second-degree Mobitz I atrioventricular (AV) block is usually associated with a disturbance at the AV nodal level, and rarely at the infra- or intra-His bundle level, it has been considered as mostly benign in character. We report on a patient with intra-His bundle block in second-degree Mobitz I AV block with right bundle branch block. PMID- 19542539 TI - Varying implantable cardioverter defibrillator referral patterns from implanting and non-implanting hospitals. AB - AIMS: To assess the impact of hospital type on implantable cardioverter defibrillation (ICD) prescription rates. METHODS AND RESULTS: The Wessex Cardiothoracic Unit is a regional implanting centre serving eight district general hospitals (DGHs). We audited all new ICD implants performed in our institution over 4 years. Hospitals implanting or referring patients elsewhere were excluded. We categorized patients into three different groups depending on local hospital type-regional centre (one hospital), DGH with a device specialist (one hospital), DGH without a device specialist (two hospitals). For each hospital type, we assessed the overall implant rate based on local population. There were 459 new ICD implants; of which 381 were included in the analysis. Implant rates were higher in areas whose local hospital was a regional centre (103/million/year), when compared with DGHs with (49/million/year) or without a device specialist (48/million/year). This disparity was greatest with respect to coronary artery disease primary prevention indications-implant rates 29, 14, and 9/million/year, respectively. CONCLUSIONS: ICD implant rates are affected by hospital type and are significantly higher in regional centres when compared with DGHs. To increase ICD implant rates, the widespread implementation of clinical pathways to identify prospective primary prevention patients may be needed. PMID- 19542540 TI - 320-detector row cardiac computed tomography in the diagnosis of arrhythmogenic right ventricular cardiomyopathy. PMID- 19542541 TI - Polymorphisms in cell death pathway genes are associated with altered sperm apoptosis and poor semen quality. AB - BACKGROUND: The FAS/FASLG system has been proposed to play a key role in germ cell apoptosis. To elucidate the role of the genetic variants of cell death pathway genes in male infertility, we genotyped the polymorphisms of FAS, FASLG and caspase-8 (CASP8) genes and evaluated their effects on sperm apoptosis and semen quality in infertile men. METHODS: The genotypes of FAS, FASLG and CASP8 were determined in 620 infertile men. Sperm apoptosis rates were measured by terminal deoxynucleotidyl transferase (Tdt)-mediated dUTP nick end labelling (TUNEL) assay, and the semen quality analysis was performed using computer assisted sperm analysis. RESULTS: We found that polymorphisms of FAS-670A/G (rs1800682: A>G) and CASP8-652 6N ins/del (rs3834129: -/CTTACT) were associated with sperm apoptosis and semen quality. Individuals with FAS-670GG showed low apoptosis rate and decreased sperm concentration, compared with the FAS-670AA genotype. Similarly, in comparison with the CASP8-652 6N ins/ins genotype, the CASP8-652 6N (ins/del+del/del) genotypes also showed significantly decreased sperm apoptosis rate and poor sperm motility. Other polymorphisms investigated did not appear to affect sperm apoptosis and semen quality. CONCLUSIONS: This study showed for the first time that functional variants of FAS and CASP8 might contribute to the dysfunctional apoptotic mechanism in germ cells, which could result in poor semen quality of ejaculated sperm. PMID- 19542542 TI - Is there a uniform basal endometrial gene expression profile during the implantation window in women who became pregnant in a subsequent ICSI cycle? AB - BACKGROUND: To understand which genes are really involved in the implantation process, we planned to study the gene basal expression profile during the window of implantation (WOI) of patients who became pregnant in a subsequent ICSI cycle. METHODS: Women attending their first ICSI cycle at ANDROS Day Surgery for severe male factor infertility were included in the study. An endometrial biopsy was performed during the WOI, in one of the last two cycles before the ICSI cycle. Forty-seven selected gene profiles were analyzed using Low Density Array technology. Only biopsies from women who subsequently became pregnant were evaluated, to exclude any bias regarding embryo viability and embryo transfer difficulties. RESULTS: Fifteen patients were included in the analysis as they became pregnant after ICSI procedure. Four of 47 selected genes were excluded from the analysis. Of the 43 genes analyzed, only 6 genes (VEGFA, PLA2G2A, ALPL, LIF, NNMT and STC1) showed a statistically uniform expression among patients who subsequently became pregnant. For all the other genes analyzed there were considerable differences in their expression levels amongst women who subsequently became pregnant. CONCLUSIONS: Our data suggest that very few genes, which change their expression level during the WOI, show a quantitative homogeneous expression in endometrially-receptive patients. In conclusion, in this study only six genes showed a homogeneous expression, and are probably involved in embryo implantation mechanisms. PMID- 19542543 TI - Expression profiling of DNA repair genes in human oocytes and blastocysts using microarrays. AB - BACKGROUND: The early preimplantation embryo relies on mRNA and protein from the oocyte to detect DNA damage and activate DNA repair, cell cycle arrest or apoptosis. Expression of some repair genes has been detected in mammalian oocytes and embryos; however, little is known about DNA repair gene expression in human blastocysts. In this study, DNA repair gene expression was investigated in human oocytes and blastocysts to identify the pathways involved at these stages and detect potential differences in repair mechanisms pre- and post-embryonic genome activation. METHODS: Triplicate sets of pooled metaphase II oocytes or blastocysts were processed for analysis using the Human Genome Survey Microarrays V2.0 (Applied Biosystems). RESULTS: Of 154 DNA repair genes investigated, 109 were detected in blastocysts and 107 in oocytes. Among differentially expressed DNA repair genes, 40/55 (73%) had lower expression levels in blastocysts compared with oocytes (P < 0.05, fold change >3). CONCLUSION: Despite experimental limitations due to culture or freezing and thawing of samples, large numbers of repair genes were detected indicating that all DNA repair pathways are potentially functional in human oocytes and blastocysts. The higher mRNA level for most repair genes in oocytes compared with blastocysts ensures sufficient availability of template until embryonic genome activation. PMID- 19542544 TI - International estimates on infertility prevalence and treatment seeking: potential need and demand for medical care. PMID- 19542546 TI - MATER protein as substrate of PKCepsilon in human cumulus cells. AB - High activity of the phosphoinositide 3-kinase/Akt pathway in cumulus cells plays an important role in FSH regulation of cell function and Protein Kinase C epsilon (PKCepsilon) collaborates with these signalling pathways to regulate cell proliferation. Relevant roles in follicular development are played by Maternal Antigen That Embryos Require (MATER) that is a cumulus cell- and oocyte-specific protein dependent on the maternal genome. We recently demonstrated that human MATER localizes at specific domains of oocytes and, for the first time, also in cumulus cells. MATER contains a carboxy-terminal leucine-rich repeat domain involved in protein-protein interactions regulating different cellular functions. Here we investigated the functional role of MATER. Thus, we performed coimmunoprecipitation experiments using HEK293T cells expressing human MATER; a similar approach was then followed in human cumulus/follicular cells. In MATER(+)HEK293T cells, we observed that this protein acts as a phosphorylation substrate of PKCepsilon. Western blot experiments indicate that, unlike oocytes, human cumulus cells express PKCepsilon. Immunoprecipitation and confocal analysis suggest for the first time that MATER protein interacts with this protein kinase in cumulus cells under physiological conditions. Since PKCepsilon is known to collaborate with antiapoptotic signalling pathways, this suggests a novel mechanism for the function of MATER in follicular maturation. PMID- 19542545 TI - Regulation of HSD1 in seeds of Arabidopsis thaliana. AB - The hydroxysteroid dehydrogenase HSD1, identified in the proteome of oil bodies from mature Arabidopsis seeds, is encoded by At5g50600 and At5g50700, two gene copies anchored on a duplicated region of chromosome 5. Using a real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) approach, the accumulation of HSD1 mRNA was shown to be specifically and highly induced in oil-accumulating tissues of maturing seeds. HSD1 mRNA disappeared during germination. The activity of HSD1 promoter and the localization of HSD1 transcripts by in situ hybridization were consistent with this pattern. A complementary set of molecular and genetic analyses showed that HSD1 is a target of LEAFY COTYLEDON2, a transcriptional regulator able to bind the promoter of HSD1. Immunoblot analyses and immunolocalization experiments using anti-AtHSD1 antibodies established that the pattern of HSD1 deposition faithfully reflected mRNA accumulation. At the subcellular level, the study of HSD1:GFP fusion proteins showed the targeting of HSD1 to the surface of oil bodies. Transgenic lines overexpressing HSD1 were then obtained to test the importance of proper transcriptional regulation of HSD1 in seeds. Whereas no impact on oil accumulation could be detected, transgenic seeds exhibited lower cold and light requirements to break dormancy, germinate and mobilize storage lipids. Interestingly, overexpressors of HSD1 over-accumulated HSD1 protein in seeds but not in vegetative organs, suggesting that post-transcriptional regulations exist that prevent HSD1 accumulation in tissues deprived of oil bodies. PMID- 19542548 TI - Enemies or allies? The organ transplant medical community, the federal government, and the public in the United States, 1967-2000. AB - The transplant medical community in the United States has frequently been divided over the appropriate role of the federal government and of the public in matters related to organ transplantation. Using public statements in government hearings, newspapers, and press releases, this article traces the thinking of the transplant medical community in particular during three especially politicized periods: the heart transplant and brain death controversies in the late 1960s, consideration of the National Organ Transplant Act and other legislation during the mid-1980s, and the controversy over organ allocation regulations issued by the Department of Health and Human Services in the late 1990s. Even while sometimes denouncing "politicization," over time surgeons, physicians, representatives of the United Network for Organ Sharing, and other leaders in the field became increasingly politically active and more accustomed to the notion that because of the unique nature of organ transplantation, both the public and the federal government have a legitimate and potentially beneficial oversight role. PMID- 19542547 TI - Dexmedetomidine, an alpha2-adrenergic agonist, inhibits neuronal delayed rectifier potassium current and sodium current. AB - BACKGROUND: Dexmedetomidine (DEX), a selective agonist of alpha2-adrenergic receptors, is recognized to facilitate analgesia and anaesthesia in humans. Despite the potential for wide use, its effects on ion currents and membrane potential in neurones remain largely unclear. METHODS: We investigated the effects of DEX on ion channels in NG108-15 neuronal cells differentiated with dibutyryl cyclic AMP and in cultured cerebellar neurones. RESULTS: DEX suppressed the amplitude of delayed rectifier K+ current [I(K(DR))] in a concentration dependent manner with an IC50 value of 4.6 microM in NG108-15 cells. No change in the steady-state inactivation of I(K(DR)) was evident in the presence of DEX. A minimal binding scheme was also used to evaluate DEX-induced block of I(K(DR)). Inhibition of I(K(DR)) by DEX was still observed in cells preincubated with yohimbine (10 microM) or efaroxan (10 microM). DEX depressed the peak amplitude of Na+ current (I(Na)), whereas it had minimal effect on L-type Ca2+ current. Under current-clamp configuration, DEX increased the duration of action potentials (APs). I(K(DR)) and I(Na) in response to AP waveforms were more sensitive to block by DEX than those elicited during rectangular pulses. In isolated cerebellar granule cells, DEX also effectively suppressed I(K(DR)). CONCLUSIONS: The effects of DEX are not limited to its interactions with alpha2 adrenergic receptors. Inhibitory effects on I(K(DR)) and I(Na) constitute one of the underlying mechanisms through which DEX and its structurally related compounds might affect neuronal activity in vivo. PMID- 19542550 TI - Deaths of neonatal calves common in Scotland in March. PMID- 19542549 TI - Health and social care responses to the Department of Health Heatwave Plan. AB - BACKGROUND: The increasing risk of heatwaves in England poses a particular threat to the health of elderly people. A National Heatwave Plan has been produced to ensure that adaptation plans are established. The objective was to explore the perceptions of frontline statutory and voluntary sector staff on the feasibility of implementing the Heatwave Plan for elderly people in the community. METHODS: Semi-structured interviews and focus groups with 109 health, social care and voluntary staff from three London Boroughs. RESULTS: Few frontline staff were aware of the Plan. Most respondents did not perceive heatwaves to be a sufficiently frequent event to require prioritization within their routine summer workloads. They highlighted the complexities associated with defining vulnerability and identifying vulnerable individuals as well as barriers to implementation of the Plan. Respondents suggested a multi-faceted approach to interventions including a public health campaign, community engagement and increasing the responsiveness of statutory services. CONCLUSION: The issues highlighted could hinder effective implementation of the Heatwave Plan. Ensuring continuity of care so that timely information can be recorded and disseminated may address the problems associated with shifting vulnerability. Best practice with respect to inter-sectoral collaboration should be identified and innovative multi-faceted interventions should be designed and evaluated. PMID- 19542551 TI - Congenital tremor and hypomyelination associated with bovine viral diarrhoea virus in 23 British cattle herds. AB - This paper presents data from 23 British herds investigated between 1991 and 2007 where neurological disease in calves was caused by bovine viral diarrhoea virus (BVDV) infection. A variety of clinical signs, most commonly tremor or trembling, were apparent in the calves from birth, and most were recumbent or unable to stand unsupported. Severe diffuse neuraxial hypomyelination was present in all of the calves, and immunohistochemistry revealed cerebral neuronal labelling consistent with congenital persistent pestivirus infection in each brain. BVDV was detected in peripheral blood samples from eight of 15 calves tested using an antigen ELISA, and was isolated in culture from samples of viscera, brain or blood collected from 17 of 24 affected calves. TaqMan RT-PCR for pestivirus RNA was positive for BVDV-1 in all six calves tested. Six of the virus isolates on which molecular classification was carried out, obtained from calves in four of the herds, were identified as BVDV-1a, while three isolates from one affected and two unaffected calves on a fifth farm were confirmed as BVDV-1b. PMID- 19542553 TI - Effects of administration of an avirulent live vaccine of Lawsonia intracellularis on mares and foals. PMID- 19542552 TI - Secondary immune-mediated thrombocytopenia in dogs naturally infected by Leishmania infantum. AB - Forty-four dogs naturally infected by Leishmania infantum were divided into two groups: 20 thrombocytopenic dogs with fewer than 150 x 10(9) platelets/l, and 24 non-thrombocytopenic dogs with more than 200 x 10(9) platelets/l. Ten clinically healthy dogs were used as controls. A haematological profile was obtained and the dogs' serum was used to assess the presence of platelet-binding IgM and IgG antibodies using a flow cytometry technique. Nineteen of the 20 thrombocytopenic dogs, and 13 of the 24 non-thrombocytopenic dogs had detectable levels of platelet-binding immunoglobulins, but none of the control dogs did so. The differences were significantly different for both IgM and IgG platelet-binding antibodies. PMID- 19542554 TI - Abomasal end-to-end anastomosis as treatment for abomasal fistulation and herniation in a cow. PMID- 19542555 TI - Use of antimicrobials. PMID- 19542556 TI - Use of antimicrobials. PMID- 19542557 TI - Echinococcus multilocularis and possible cycles in UK wildlife. PMID- 19542558 TI - Bovine tuberculosis in Eurasian otters. PMID- 19542559 TI - Proposed veterinary union. PMID- 19542560 TI - NIH peer review reform. PMID- 19542561 TI - Proline-rich sequence recognition: II. Proteomics analysis of Tsg101 ubiquitin-E2 like variant (UEV) interactions. AB - The tumor maintenance protein Tsg101 has recently gained much attention because of its involvement in endosomal sorting, virus release, cytokinesis, and cancerogenesis. The ubiquitin-E2-like variant (UEV) domain of the protein interacts with proline-rich sequences of target proteins that contain P(S/T)AP amino acid motifs and weakly binds to the ubiquitin moiety of proteins committed to sorting or degradation. Here we performed peptide spot analysis and phage display to refine the peptide binding specificity of the Tsg101 UEV domain. A mass spectrometric proteomics approach that combines domain-based pulldown experiments, binding site inactivation, and stable isotope labeling by amino acids in cell culture (SILAC) was then used to delineate the relative importance of the peptide and ubiquitin binding sites. Clearly "PTAP" interactions dominate target recognition, and we identified several novel binders as for example the poly(A)-binding protein 1 (PABP1), Sec24b, NFkappaB2, and eIF4b. For PABP1 and eIF4b the interactions were confirmed in the context of the corresponding full length proteins in cellular lysates. Therefore, our results strongly suggest additional roles of Tsg101 in cellular regulation of mRNA translation. Regulation of Tsg101 itself by the ubiquitin ligase TAL (Tsg101-associated ligase) is most likely conferred by a single PSAP binding motif that enables the interaction with Tsg101 UEV. Together with the results from the accompanying article (Kofler, M., Schuemann, M., Merz, C., Kosslick, D., Schlundt, A., Tannert, A., Schaefer, M., Luhrmann, R., Krause, E., and Freund, C. (2009) Proline-rich sequence recognition: I. Marking GYF and WW domain assembly sites in early spliceosomal complexes. Mol. Cell. Proteomics 8, 2461-2473) on GYF and WW domain pathways our work defines major proline-rich sequence-mediated interaction networks that contribute to the modular assembly of physiologically relevant protein complexes. PMID- 19542563 TI - Solution structure of human cardiac troponin C in complex with the green tea polyphenol, (-)-epigallocatechin 3-gallate. AB - Heart muscle contraction is regulated by Ca(2+) binding to the thin filament protein troponin C. In cardiovascular disease, the myofilament response to Ca(2+) is often altered. Compounds that rectify this perturbation are of considerable interest as therapeutics. Plant flavonoids have been found to provide protection against a variety of human illnesses such as cancer, infection, and heart disease. (-)-Epigallocatechin gallate (EGCg), the prevalent flavonoid in green tea, modulates force generation in isolated guinea pig hearts (Hotta, Y., Huang, L., Muto, T., Yajima, M., Miyazeki, K., Ishikawa, N., Fukuzawa, Y., Wakida, Y., Tushima, H., Ando, H., and Nonogaki, T. (2006) Eur. J. Pharmacol. 552, 123-130) and in skinned cardiac muscle fibers (Liou, Y. M., Kuo, S. C., and Hsieh, S. R. (2008) Pflugers Arch. 456, 787-800; and Tadano, N., Yumoto, F., Tanokura, M., Ohtsuki, I., and Morimoto, S. (2005) Biophys. J. 88, 314a). In this study we describe the solution structure of the Ca(2+)-saturated C-terminal domain of troponin C in complex with EGCg. Moreover, we show that EGCg forms a ternary complex with the C-terminal domain of troponin C and the anchoring region of troponin I. The structural evidence indicates that the binding site of EGCg on the C-terminal domain of troponin C is in the hydrophobic pocket in the absence of troponin I, akin to EMD 57033. Based on chemical shift mapping, the binding of EGCg to the C-terminal domain of troponin C in the presence of troponin I may be to a new site formed by the troponin C.troponin I complex. This interaction of EGCg with the C-terminal domain of troponin C.troponin I complex has not been shown with other cardiotonic molecules and illustrates the potential mechanism by which EGCg modulates heart contraction. PMID- 19542564 TI - Chronic kidney disease delays VLDL-apoB-100 particle catabolism: potential role of apolipoprotein C-III. AB - To determine the relative contribution of obesity and/or insulin resistance (IR) in the development of dyslipidemia in chronic kidney disease (CKD), we investigated the transport of apolipoprotein (apo) B-100 in nonobese, nondiabetic, nonnephrotic CKD subjects and healthy controls (HC). We determined total VLDL, VLDL(1), VLDL(2), intermediate density lipoprotein (IDL), and LDL apoB-100 using intravenous D3-leucine, GC-MS, and multicompartmental modeling. Plasma apoC-III and apoB-48 were immunoassayed. In this case control study, we report higher plasma triglyceride, IDL-, VLDL-, VLDL(1)-, and VLDL(2)-apoB-100 concentrations in CKD compared with HC (P < 0.05). This was associated with decreased fractional catabolic rates [FCRs (pools/day)] [IDL:CKD 3.4 (1.6) vs. HC 5.0 (3.2), P < 0.0001; VLDL:CKD 4.8 (5.2) vs. HC 7.8 (4.8), P = 0.038; VLDL(1):CKD 10.1 (8.5) vs. HC 29.5 (45.1), P = 0.007; VLDL(2):CKD 5.4 (4.6) vs. HC 10.4 (3.4), P = 0.001] with no difference in production rates. Plasma apoC-III and apoB-48 were significantly higher in CKD (P < 0.001) and both correlated with impaired FCRs of VLDL, VLDL(1), and VLDL(2) apoB-100 (P < 0.05). In CKD, apoC-III concentration was the only independent predictor of clearance defects in VLDL and its subfractions. Moderate CKD in the absence of central adiposity and IR is associated with mild hypertriglyceridemia due to delayed catabolism of triglyceride rich lipoproteins, IDL, and VLDL, without changes in production rate. Altered apoC-III metabolism may contribute to dyslipidemia in CKD, and this requires further investigation. PMID- 19542562 TI - Mechanical regulation of the proangiogenic factor CCN1/CYR61 gene requires the combined activities of MRTF-A and CREB-binding protein histone acetyltransferase. AB - Smooth muscle-rich tissues respond to mechanical overload by an adaptive hypertrophic growth combined with activation of angiogenesis, which potentiates their mechanical overload-bearing capabilities. Neovascularization is associated with mechanical strain-dependent induction of angiogenic factors such as CCN1, an immediate-early gene-encoded matricellular molecule critical for vascular development and repair. Here we have demonstrated that mechanical strain dependent induction of the CCN1 gene involves signaling cascades through RhoA mediated actin remodeling and the p38 stress-activated protein kinase (SAPK). Actin signaling controls serum response factor (SRF) activity via SRF interaction with the myocardin-related transcriptional activator (MRTF)-A and tethering to a single CArG box sequence within the CCN1 promoter. Such activity was abolished in mechanically stimulated mouse MRTF-A(-/-) cells or upon inhibition of CREB binding protein (CBP) histone acetyltransferase (HAT) either pharmacologically or by siRNAs. Mechanical strain induced CBP-mediated acetylation of histones 3 and 4 at the SRF-binding site and within the CCN1 gene coding region. Inhibition of p38 SAPK reduced CBP HAT activity and its recruitment to the SRF.MRTF-A complex, whereas enforced induction of p38 by upstream activators (e.g. MKK3 and MKK6) enhanced both CBP HAT and CCN1 promoter activities. Similarly, mechanical overload-induced CCN1 gene expression in vivo was associated with nuclear localization of MRTF-A and enrichment of the CCN1 promoter with both MRTF-A and acetylated histone H3. Taken together, these data suggest that signal-controlled activation of SRF, MRTF-A, and CBP provides a novel connection between mechanical stimuli and angiogenic gene expression. PMID- 19542565 TI - GPIHBP1 stabilizes lipoprotein lipase and prevents its inhibition by angiopoietin like 3 and angiopoietin-like 4. AB - Glycosylphosphatidylinositol-anchored HDL-binding protein (GPIHBP1) binds both LPL and chylomicrons, suggesting that GPIHBP1 is a platform for LPL-dependent processing of triglyceride (TG)-rich lipoproteins. Here, we investigated whether GPIHBP1 affects LPL activity in the absence and presence of LPL inhibitors angiopoietin-like (ANGPTL)3 and ANGPTL4. Like heparin, GPIHBP1 stabilized but did not activate LPL. ANGPTL4 potently inhibited nonstabilized LPL as well as heparin stabilized LPL but not GPIHBP1-stabilized LPL. Like ANGPTL4, ANGPTL3 inhibited nonstabilized LPL but not GPIHBP1-stabilized LPL. ANGPTL3 also inhibited heparin stabilized LPL but with less potency than nonstabilized LPL. Consistent with these in vitro findings, fasting serum TGs of Angptl4(-/-)/Gpihbp1(-/-) mice were lower than those of Gpihbp1(-/-) mice and approached those of wild-type littermates. In contrast, serum TGs of Angptl3(-/-)/Gpihbp1(-/-) mice were only slightly lower than those of Gpihbp1(-/-) mice. Treating Gpihbp1(-/-) mice with ANGPTL4- or ANGPTL3-neutralizing antibodies recapitulated the double knockout phenotypes. These data suggest that GPIHBP1 functions as an LPL stabilizer. Moreover, therapeutic agents that prevent LPL inhibition by ANGPTL4 or, to a lesser extent, ANGPTL3, may benefit individuals with hyperlipidemia caused by gene mutations associated with decreased LPL stability. PMID- 19542566 TI - ["Risk factors for childhood depression"--research design, implementation, proceedings: history of 13 years: experience in grant preparation, writing, organization in connection with an American NIMH Grant]. AB - The authors summarize their experiences in research organization accumulated during 13 years. At first they outline preliminary studies which are prerequisites of high prestige international grants. Then they describe the huge administrative apparatus dedicated - besides skilled professionals - for the construction and organization of the research, the management, continuous checking and evaluation of data in such a multisite study. Finally, they report on the scientific results obtained after 13 years of hard work. PMID- 19542567 TI - [Hungarian translation of the Dimensional Yale-Brown Obsessive-Compulsive Scale and our first experiences with the test]. AB - According to the dimensional approach of the obsessive-compulsive disorder (OCD), it is possible to identify phenotypic subtypes--so called dimensions--within the heterogeneous clinical manifestations of OCD by statistical techniques (principal component analysis). The Dimensional Yale-Brown Obsessive Compulsive Scale (DY BOCS) measures the presence of the obsessive-compulsive symptoms within the phenotype. The scale is a semi-structured instrument, which includes a symptom list divided into six dimensions for the patients and a part reserved for the clinician. The authors gave their written permission for the translation of the DY-BOCS test into Hungarian, revised the back-translation of the Hungarian version and the final, revised version is published for the first time, in Hungary. The Hungarian DY-BOCS was administered to seventeen OCD patients at 2nd Department of Psychiatry, Nyiro Gyula Hospital. The reliability analysis produced strong correlation coefficients. The internal consistency (Cronbach alpha) scores were high, ranging from 0.97 to 0.99. The scores of the validity scales were also good. There was a strong convergent validity between the gold standard, Yale Brown Obsessive-Compulsive Scale (Y-BOCS) global severity scores and the DY-BOCS global severity scores (p=0.001 / r=0.744). The examination of the internal validity showed that the DY-BOCS global score and the aggressive and miscellaneous dimension severity rating scores are strongly correlated. We also experienced strong correlation between the DY-BOCS global score and the component scores of the global severity ratings (time, distress, interference, and impairment), the Pearson's r ranged from 0.815 to 0.960. There was no significant inter-correlation between the dimensions, which supports the assumption that these dimensions are independent constructs. In the divergent validity measures, we used the Hamilton Depression Scale (HAM-D) to compare OCD dimensions with the most common comorbid disorder (depression) symptoms. The severity scores of the HAM-D were correlated with the severity scores of the DY-BOCS dimensions. In this latter validity test, we found only one significant correlation between the aggressive dimension and the severity of the depression. In the case of the remaining dimensions, there was no significant correlation between the dimensions and the severity of depression; thus, it seems that the two tests examine widely different psychological phenomena . The evaluating scores of the Hungarian version of the DY-BOCS are very close to the original published scores. These results indicate that the Hungarian version of the DY-BOCS is a reliable and a valid clinical tool and we hope that by using the test, Hungarian professionals will become familiar with and accept the dimensional approach of OCD. PMID- 19542568 TI - [The importance and possibilities of cognitive reconstructing in the therapy of pathological gambling]. AB - Cognitive reconstructing is a major component of cognitive and cognitive behavioural therapies. Cognitive reconstructing means identification and modification of cognitive distortions. Research into this field has shown that cognitive distortions about gambling are clearly implicated in the development and persistence of pathological gambling. This paper outlines the use of cognitive reconstructing for the therapy of pathological gambling. It highlights the type of cognitive distortions most common to this population and outlines the role of semistructured interviews as tool for exploring these distortions. Modification of these beliefs with Socratic dialog is also discussed in the paper. Finally, it describes the studies conducted so far to assess the therapeutic efficacy of cognitive reconstructing in treating pathological gambling. PMID- 19542569 TI - [Associations between symptom severity and emotion recognition in depressed patients]. AB - INTRODUCTION: In this study we tried to explore whether there are any associations between symptom severity and emotional facial expression recognition. Based on the literature and our earlier findings, we believe that the impairments of overall (not emotion specific) facial expression recognition is associated with a more general, non-specific psychiatric distress. Thus, first it was hypothesized that overall facial expression recognition would be correlated negatively with SCL-90 global severity index (GSI). Our second hypothesis was that the severity of the impairment in facial expression recognition would be correlated with symptom severity measured by the BECK Depression Inventory (BDI). METHODS: One hundred and six depressed subjects participated in the study. We used a 'Virtual Human' for presenting the 35 emotional and neutral facial expressions. Subjects filled in the BDI and SCL-90 questionnaires after the facial expression recognition task. RESULTS: Significant negative association was found between the GSI of SCL-90 and the overall facial expression recognition. Furthermore, significant negative correlation was found between the recognition of happy facial expressions and the depression severity measured by the BDI questionnaire and the SCL-90 depression subscale. DISCUSSION: The negative association found between overall facial expression recognition and the GSI score provide empirical support to our first hypothesis, which states that the impairment of not emotion specific, overall facial expression recognition may correlate with a general psychiatric distress. Our findings indicate that those depressed subjects who had higher scores on BDI could recognize happy facial expression at a significantly lower rate. Abnormalities in reward sensitivity may explain this association between happy face recognition difficulties and the severity of depressive symptoms among depressive patients. Consequently, we can conclude that the recognition of happy facial expressions can be a useful tool for monitoring the severity of depression. PMID- 19542570 TI - [The relationship between burnout and religious belief among nuns serving as nurses]. AB - In this paper the findings of a study designed to assess whether nuns serving as nurses and having definite religious persuasions feel the same degree of burnout in their service of helping patients as non-religious nurses are presented. For this purpose two questionnaires were used, namely the Post-Critical Belief Scale and the Maslach Burnout Inventory. Subjects of the study had been working as a nurse for at least 5 years and 188 such subjects were enrolled. The causative factors behind burnout were studied. Only women were included in the study. Statistical analysis has surprisingly shown that the extent of burnout did not increase in direct proportion with the years spent at work and burnout as a dependent variable could be explained by age only to a minimal extent. After this, the influence of another factor, i.e. religious belief, on burnout was studied. However, religious belief cannot be described and quantified with a single numerical value; therefore, three aspects were measured. These aspects included the self-rated degree of religious belief, attendance at religious services and the "Post-Critical Belief Scale". The values based on these aspects showed a linear relationship and were indirectly proportional with burnout. Consequently, religious belief can be regarded as a protective factor against burnout in the present sample. In addition, the extent of burnout was found to be the lowest in the community of nuns living in a religious order, it was higher among nurses working at religious institutions and it was the highest among nurses working at state hospitals. Thus, religious belief has a decisive role in avoiding a burnout. PMID- 19542571 TI - Efficient sparse kernel feature extraction based on partial least squares. AB - The presence of irrelevant features in training data is a significant obstacle for many machine learning tasks. One approach to this problem is to extract appropriate features and, often, one selects a feature extraction method based on the inference algorithm. Here, we formalize a general framework for feature extraction, based on Partial Least Squares, in which one can select a user defined criterion to compute projection directions. The framework draws together a number of existing results and provides additional insights into several popular feature extraction methods. Two new sparse kernel feature extraction methods are derived under the framework, called Sparse Maximal Alignment (SMA) and Sparse Maximal Covariance (SMC), respectively. Key advantages of these approaches include simple implementation and a training time which scales linearly in the number of examples. Furthermore, one can project a new test example using only k kernel evaluations, where k is the output dimensionality. Computational results on several real-world data sets show that SMA and SMC extract features which are as predictive as those found using other popular feature extraction methods. Additionally, on large text retrieval and face detection data sets, they produce features which match the performance of the original ones in conjunction with a Support Vector Machine. PMID- 19542572 TI - Watershed cuts: minimum spanning forests and the drop of water principle. AB - We study the watersheds in edge-weighted graphs. We define the watershed cuts following the intuitive idea of drops of water flowing on a topographic surface. We first establish the consistency of these watersheds: They can be equivalently defined by their "catchment basins" (through a steepest descent property) or by the "dividing lines" separating these catchment basins (through the drop of water principle). Then, we prove, through an equivalence theorem, their optimality in terms of minimum spanning forests. Afterward, we introduce a linear-time algorithm to compute them. To the best of our knowledge, similar properties are not verified in other frameworks and the proposed algorithm is the most efficient existing algorithm, both in theory and in practice. Finally, the defined concepts are illustrated in image segmentation, leading to the conclusion that the proposed approach improves, on the tested images, the quality of watershed-based segmentations. PMID- 19542573 TI - Appearance derivatives for isonormal clustering of scenes. AB - A new technique is proposed for scene analysis, called "appearance clustering." The key result of this approach is that the scene points can be clustered according to their surface normals, even when the geometry, material, and lighting are all unknown. This is achieved by analyzing an image sequence of a scene as it is illuminated by a smoothly moving distant light source. In such a scenario, the brightness measurements at each pixel form a "continuous appearance profile." When the source path follows an unstructured trajectory (obtained, say, by smoothly hand-waving a light source), the locations of the extrema of the appearance profile provide a strong cue for the scene point's surface normal. Based on this observation, a simple transformation of the appearance profiles and a distance metric are introduced that, together, can be used with any unsupervised clustering algorithm to obtain isonormal clusters of a scene. We support our algorithm empirically with comprehensive simulations of the Torrance Sparrow and Oren-Nayar analytic BRDFs, as well as experiments with 25 materials obtained from the MERL database of measured BRDFs. The method is also demonstrated on 45 examples from the CURET database, obtaining clusters on scenes with real textures such as artificial grass and ceramic tile, as well as anisotropic materials such as satin and velvet. The results of applying our algorithm to indoor and outdoor scenes containing a variety of complex geometry and materials are shown. As an example application, isonormal clusters are used for lighting-consistent texture transfer. Our algorithm is simple and does not require any complex lighting setup for data collection. PMID- 19542575 TI - Automatic writer identification of ancient Greek inscriptions. AB - This paper introduces a novel methodology for the classification of ancient Greek inscriptions according to the writer who carved them. Inscription writer identification is crucial for dating the written content, which in turn is of fundamental importance in the sciences of history and archaeology. To achieve this, we first compute an ideal or "platonic" prototype for the letters of each inscription separately. Next, statistical criteria are introduced to reject the hypothesis that two inscriptions are carved by the same writer. In this way, we can determine the number of distinct writers who carved a given ensemble of inscriptions. Next, maximum likelihood considerations are employed to attribute all inscriptions in the collection to the respective writers. The method has been applied to 24 Ancient Athenian inscriptions and attributed these inscriptions to six different identified hands in full accordance with expert epigraphists' opinions. PMID- 19542574 TI - Automatic summarization of changes in biological image sequences using algorithmic information theory. AB - An algorithmic information-theoretic method is presented for object-level summarization of meaningful changes in image sequences. Object extraction and tracking data are represented as an attributed tracking graph (ATG). Time courses of object states are compared using an adaptive information distance measure, aided by a closed-form multidimensional quantization. The notion of meaningful summarization is captured by using the gap statistic to estimate the randomness deficiency from algorithmic statistics. The summary is the clustering result and feature subset that maximize the gap statistic. This approach was validated on four bioimaging applications: 1) It was applied to a synthetic data set containing two populations of cells differing in the rate of growth, for which it correctly identified the two populations and the single feature out of 23 that separated them; 2) it was applied to 59 movies of three types of neuroprosthetic devices being inserted in the brain tissue at three speeds each, for which it correctly identified insertion speed as the primary factor affecting tissue strain; 3) when applied to movies of cultured neural progenitor cells, it correctly distinguished neurons from progenitors without requiring the use of a fixative stain; and 4) when analyzing intracellular molecular transport in cultured neurons undergoing axon specification, it automatically confirmed the role of kinesins in axon specification. PMID- 19542576 TI - Canonical correlation analysis of video volume tensors for action categorization and detection. AB - This paper addresses a spatiotemporal pattern recognition problem. The main purpose of this study is to find a right representation and matching of action video volumes for categorization. A novel method is proposed to measure video-to video volume similarity by extending Canonical Correlation Analysis (CCA), a principled tool to inspect linear relations between two sets of vectors, to that of two multiway data arrays (or tensors). The proposed method analyzes video volumes as inputs avoiding the difficult problem of explicit motion estimation required in traditional methods and provides a way of spatiotemporal pattern matching that is robust to intraclass variations of actions. The proposed matching is demonstrated for action classification by a simple Nearest Neighbor classifier. We, moreover, propose an automatic action detection method, which performs 3D window search over an input video with action exemplars. The search is speeded up by dynamic learning of subspaces in the proposed CCA. Experiments on a public action data set (KTH) and a self-recorded hand gesture data showed that the proposed method is significantly better than various state-of-the-art methods with respect to accuracy. Our method has low time complexity and does not require any major tuning parameters. PMID- 19542577 TI - A hybrid feature extraction selection approach for high-dimensional non-Gaussian data clustering. AB - This paper presents an unsupervised approach for feature selection and extraction in mixtures of generalized Dirichlet (GD) distributions. Our method defines a new mixture model that is able to extract independent and non-Gaussian features without loss of accuracy. The proposed model is learned using the Expectation Maximization algorithm by minimizing the message length of the data set. Experimental results show the merits of the proposed methodology in the categorization of object images. PMID- 19542579 TI - On symmetry, perspectivity, and level-set-based segmentation. AB - We introduce a novel variational method for the extraction of objects with either bilateral or rotational symmetry in the presence of perspective distortion. Information on the symmetry axis of the object and the distorting transformation is obtained as a by--product of the segmentation process. The key idea is the use of a flip or a rotation of the image to segment as if it were another view of the object. We call this generated image the symmetrical counterpart image. We show that the symmetrical counterpart image and the source image are related by planar projective homography. This homography is determined by the unknown planar projective transformation that distorts the object symmetry. The proposed segmentation method uses a level-set-based curve evolution technique. The extraction of the object boundaries is based on the symmetry constraint and the image data. The symmetrical counterpart of the evolving level-set function provides a dynamic shape prior. It supports the segmentation by resolving possible ambiguities due to noise, clutter, occlusions, and assimilation with the background. The homography that aligns the symmetrical counterpart to the source level-set is recovered via a registration process carried out concurrently with the segmentation. Promising segmentation results of various images of approximately symmetrical objects are shown. PMID- 19542578 TI - A novel knowledge-based system for interpreting complex engineering drawings: theory, representation, and implementation. AB - We present a novel knowledge-based system to automatically convert real-life engineering drawings to content-oriented high-level descriptions. The proposed method essentially turns the complex interpretation process into two parts: knowledge representation and knowledge-based interpretation. We propose a new hierarchical descriptor-based knowledge representation method to organize the various types of engineering objects and their complex high-level relations. The descriptors are defined using an Extended Backus Naur Form (EBNF), facilitating modification and maintenance. When interpreting a set of related engineering drawings, the knowledge-based interpretation system first constructs an EBNF-tree from the knowledge representation file, then searches for potential engineering objects guided by a depth-first order of the nodes in the EBNF-tree. Experimental results and comparisons with other interpretation systems demonstrate that our knowledge-based system is accurate and robust for high-level interpretation of complex real-life engineering projects. PMID- 19542580 TI - Probabilistic modeling of scene dynamics for applications in visual surveillance. AB - We propose a novel method to model and learn the scene activity, observed by a static camera. The proposed model is very general and can be applied for solution of a variety of problems. The motion patterns of objects in the scene are modeled in the form of a multivariate nonparametric probability density function of spatiotemporal variables (object locations and transition times between them). Kernel Density Estimation is used to learn this model in a completely unsupervised fashion. Learning is accomplished by observing the trajectories of objects by a static camera over extended periods of time. It encodes the probabilistic nature of the behavior of moving objects in the scene and is useful for activity analysis applications, such as persistent tracking and anomalous motion detection. In addition, the model also captures salient scene features, such as the areas of occlusion and most likely paths. Once the model is learned, we use a unified Markov Chain Monte Carlo (MCMC)-based framework for generating the most likely paths in the scene, improving foreground detection, persistent labeling of objects during tracking, and deciding whether a given trajectory represents an anomaly to the observed motion patterns. Experiments with real world videos are reported which validate the proposed approach. PMID- 19542581 TI - Synergy between object recognition and image segmentation using the expectation maximization algorithm. AB - In this work, we formulate the interaction between image segmentation and object recognition in the framework of the Expectation-Maximization (EM) algorithm. We consider segmentation as the assignment of image observations to object hypotheses and phrase it as the E-step, while the M-step amounts to fitting the object models to the observations. These two tasks are performed iteratively, thereby simultaneously segmenting an image and reconstructing it in terms of objects. We model objects using Active Appearance Models (AAMs) as they capture both shape and appearance variation. During the E-step, the fidelity of the AAM predictions to the image is used to decide about assigning observations to the object. For this, we propose two top-down segmentation algorithms. The first starts with an oversegmentation of the image and then softly assigns image segments to objects, as in the common setting of EM. The second uses curve evolution to minimize a criterion derived from the variational interpretation of EM and introduces AAMs as shape priors. For the M-step, we derive AAM fitting equations that accommodate segmentation information, thereby allowing for the automated treatment of occlusions. Apart from top-down segmentation results, we provide systematic experiments on object detection that validate the merits of our joint segmentation and recognition approach. PMID- 19542582 TI - Statistical Hough Transform. AB - The Standard Hough Transform is a popular method in image processing and is traditionally estimated using histograms. Densities modeled with histograms in high dimensional space and/or with few observations, can be very sparse and highly demanding in memory. In this paper, we propose first to extend the formulation to continuous kernel estimates. Second, when dependencies in between variables are well taken into account, the estimated density is also robust to noise and insensitive to the choice of the origin of the spatial coordinates. Finally, our new statistical framework is unsupervised (all needed parameters are automatically estimated) and flexible (priors can easily be attached to the observations). We show experimentally that our new modeling encodes better the alignment content of images. PMID- 19542584 TI - Scale-space behavior of planar-curve corners. AB - The curvature scale-space (CSS) technique is suitable for extracting curvature features from objects with noisy boundaries. To detect corner points in a multiscale framework, Rattarangsi and Chin investigated the scale-space behavior of planar-curve corners. Unfortunately, their investigation was based on an incorrect assumption, viz., that planar curves have no shrinkage under evolution. In the present paper, this mistake is corrected. First, it is demonstrated that a planar curve may shrink nonuniformly as it evolves across increasing scales. Then, by taking into account the shrinkage effect of evolved curves, the CSS trajectory maps of various corner models are investigated and their properties are summarized. The scale-space trajectory of a corner may either persist, vanish, merge with a neighboring trajectory, or split into several trajectories. The scale-space trajectories of adjacent corners may attract each other when the corners have the same concavity, or repel each other when the corners have opposite concavities. Finally, we present a standard curvature measure for computing the CSS maps of digital curves, with which it is shown that planar curve corners have consistent scale-space behavior in the digital case as in the continuous case. PMID- 19542583 TI - Exploration of shape variation using localized components analysis. AB - Localized Components Analysis (LoCA) is a new method for describing surface shape variation in an ensemble of objects using a linear subspace of spatially localized shape components. In contrast to earlier methods, LoCA optimizes explicitly for localized components and allows a flexible trade-off between localized and concise representations, and the formulation of locality is flexible enough to incorporate properties such as symmetry. This paper demonstrates that LoCA can provide intuitive presentations of shape differences associated with sex, disease state, and species in a broad range of biomedical specimens, including human brain regions and monkey crania. PMID- 19542585 TI - Piecewise linear models with guaranteed closeness to the data. AB - This paper addresses the problem of piecewise linear approximation of point sets without any constraints on the order of data points or the number of model components (line segments). We point out two problems with the maximum likelihood estimate (MLE) that present serious drawbacks in practical applications. One is that the parametric models obtained using a classical MLE framework are not guaranteed to be close to data points. It is typically impossible, in this classical framework, to detect whether a parametric model fits the data well or not. The second problem is related to accurately choosing the optimal number of model components. We first fit a nonparametric density to the data points and use it to define a neighborhood of the data. Observations inside this neighborhood are deemed informative; those outside the neighborhood are deemed uninformative for our purpose. This provides us with a means to recognize when models fail to properly fit the data. We then obtain maximum likelihood estimates by optimizing the Kullback-Leibler Divergence (KLD) between the nonparametric data density restricted to this neighborhood and a mixture of parametric models. We prove that, under the assumption of a reasonably large sample size, the inferred model components are close to their ground-truth model component counterparts. This holds independently of the initial number of assumed model components or their associated parameters. Moreover, in the proposed approach, we are able to estimate the number of significant model components without any additional computation. PMID- 19542586 TI - Comments on "what the back of the object looks like: 3D reconstruction from line drawings without hidden lines". AB - I comment on a paper describing a method for deducing the hidden topology of an object portrayed in a 2D natural line drawing. The principal problem with this paper is that it cannot be considered an advance on (or even an equal of) the state of the art as the approach it describes makes the same limiting assumptions as approaches proposed 10 years ago. There are also important omissions in the review of related work. PMID- 19542587 TI - Responses to the comments on "what the back of the object looks like: 3D reconstruction from line drawings without hidden lines". AB - Varley [1] made comments on our paper in [2] section by section. We answer them in this response paper. PMID- 19542588 TI - [Testosterone, endothelial function, cardiovascular health and androgen deficiency in the old man]. AB - Testosterone determination in an old men population has demonstrated its about the as general health marker, not only sexual, prompting a greater in to arrest for this analytic determination and the potential relations of testosterone with other markers of cardiovascular health, obesity, hypertension, erectile dysfunction, sarcopenia, metabolic syndrome, ageing, and other conditions. We specifically review the relationship between cardiovascular health, erectile dysfunction, and androgen deficiency, processes easily recognizable, prevented and treated. Current information gives such a prominence to testosterone as a health reference that its determination seems to be inexcusable in the ageing male consult. PMID- 19542589 TI - [Charles II: From spell to genitourinary pathology]. AB - OBJECTIVES: We attempt to unravel the complex condition of the last king of the Hapsburg dynasty in Spain, Charles II, called The Bewitched, in whom a genitourinary disorder was preponderant, preventing him from fulfilling one of the objectives of the monarchial institution, engendering a heir, and causing a series of complications that led to his death. METHODS: We review the works describing the life of the King, with special emphasis on his bloodline, the doubts about his sex at birth, his pathological processes, the repercussion among European Courts, but above all on his marriages and the inability to engender an heir. We also emphasize the thought of 17th century Spanish society which led to the belief that he was bewitched. The nickname he passed into history with. RESULTS: It was deduced that he could have presented posterior hypospadias which, together with monorchism and atrophic testicle, led to the belief that he presented an intersexual state with ambiguous genitals. The physical phenotype leans more towards true hermaphro-ditism and above all a XX male, rather than the more often attributed Klinefelter's syndrome. This is probably also associated with a fragile X syndrome. Very possibly congenital monorenal, death was due to chronic kidney failure caused by glomerulopathy or interstitial nephro-pathy as a consequence of renal lithiasis plus recurrent infections of the urinary tract. CONCLUSIONS: As a result of a reiterated endogamic matrimonial policy, the Hapsburg dynasty died out in Spain in 1700, represented by Charles II, a pluripatho-logical king who can only be freed from speculation by chromosomal and genetic studies of his remains buried in El Escorial monastery. PMID- 19542590 TI - [Approach of renal parenchyma infection: retrospective study of period 2005 2007]. AB - BACKGROUND: The renal parenchyma acute infection, known as acute pyelonephritis (APN), is diagnosed and treated in some Hospital Departments of the H.C.U. Lozano Blesa. We want to know if the process was made in a homogeneous way, fixed to the described quality standards and if we could detect improvement areas. METHODS: Retrospective study in admitted patients with the diagnosis of APN over and 2 year period (11-05/10-07), evaluating some variables referred to epidemiological data, diagnosis, treatment and evolution. RESULTS: We studied 118 patients (80.5% women, mean age 34 years), that supposed 0.39% of patients admitted from the Emergency Department (36% in Urology and the rest in Paediatric, Obstetric, Internal Medicine and others).45% showed risk factors (urinary anomalies, diabetes, pregnancy, immunosuppressant, ...), and it is considered complicated APN (CAPN). In children there was a high rate of urinary tract malformations. Diagnosis was clinical in 82.2%. The rest required imaging techniques, specially children, using gamma scan. In 82.7% there was leukocyturia and in 79.9% leukocytosis. A urine culture pre-treatment was made in 76.3%, with a positive rate of 55.5%, detecting negative Gram germs in 94% (E. coli in 82%). Treatment was empiric in all cases, based on cephalosporin, amoxicillin/clavulanic acid and fluoroquinilones. Evolution was favorable in 93.1% (95.1% of NCAPN and 85.7% of CAPN). The mean hospital stay was 6.4 days (5.6 in NCAPN and 7.5 in CAPN). CONCLUSIONS: Acute pyelonephritis management in our hospital is highly satisfactory and similar to the revised medical literature. We could emphasize the low rate of urinary cultures pretreatment (negative in quite a lot of the cases, due to early beginning of antibiotic treatment) and that mean hospital stay could be reduced in CAPN. We want to congratulate all involved professionals at the hospital for the good practice demonstrated. PMID- 19542591 TI - [Implantation technique of the artificial urinary sphincter "FlowSecure" in the bulbar urethra]. AB - BACKGROUND: Artificial urinary sphincter "FlowSecure" is a prosthesis designed for stress urinary incontinence that has achieved excellent results. Although implantation is easy, some urologist used to other prosthesis may find difficult the change to this new technique. This article shows how easily and quickly this new sphincter can be implanted and discuss the differences with the artificial sphincter AMS-800. METHODS: Following the case of a patient who was implanted artificial urinary sphincter "FlowSecure" in our center we describe with pictures the technique of implantation and give some advices to make this quicker and easier. RESULTS: Surgical time was 90 minutes. Patient presented uneventful recovery, urethral catheter was removed 24 hours after surgery and discharge was done 72 hours after surgery. Three months after implantation patient describes complete resolution of his stress urinary incontinence. CONCLUSIONS: Artificial urinary sphincter "FlowSecure" is easy and quick to implant, and as its use is extended we would compare if results are better than those of the AMS-800 model. PMID- 19542592 TI - [Correlation between symptoms and survival in patients with renal cell carcinoma]. AB - OBJECTIVES: To evaluate the correlation between type of presentation in patients with renal cell carcinoma and survival. METHODS: Longitudinal retrospective study of 42 patients with clinical and histological diagnosis of renal cell carcinoma (RCC) between July 2002 and September 2007 at University Hospital Manuel Fajardo. The following variables were studied: Sex, age, clinical presentation: 1) asymptomatic: 2) symptomatic: 3) paraneoplastic syndrome; tumor stage (TNM 1997) and survival. RESULTS: There is a predominance of men over women, the age of greater incidence was between the fifth a and seventh decades. The tumors were classified as: asymptomatic 7%, 3/42; symptomatic 67%, 28/42, with flank pain 31%, 13/42; with hematuria 24%, 10/42, and tumor 10%, 4/42, the classic triad of hematuria, flank pain and mass was 2.3%, 1/42 of the patients and paraneoplastic syndrome 24%, 11/42. The patients with non-metastatic disease were 57%, 24/42 and 43%, 18/42 presented metastases. Five-year survival for patients with RCC by TNM stage was T1, 100% ( 12/12) , T2, 97% (11/12) , T3, 36% (4/11) , T4, 0% (0/7) . The survival of localized and metastatic RCC was 95%, 23/24 and 22%, 4/18 cases respectively and all stage was 64% (27/42). CONCLUSIONS: The asymptomatic RCCs were smaller; pain, hematuria, and mass were the most common manifestations in symptomatic RCC. The prognosis is dismal in patients with RCC showing paraneoplastic syndromes. PMID- 19542593 TI - [Clinical presentation of renal cell carcinoma in renal transplant]. AB - OBJECTIVES: To analyze the clinical presentation and therapeutic response of renal cell carcinoma (RCC) of the renal graft. METHODS: Analysis of the cases described in our centre and review of current literature. RESULTS: RCC has a higher incidence in transplant patients, affecting the graft in less than 10% of the cases. Detection is usually a casual event during follow-up due to the absence of innervation, although its presentation may be as an acute abdomen in case of breakage of the graft. Conventional treatment consists of transplant nephrectomy, but partial nephrectomy has been performed in recent years with good results. The modification of immunosuppression is a routine measure after treatment. CONCLUSIONS: The incidence of RCC after renal transplants in our series is 0.7%, of which 22% are originated in the graft. The clinical presentation of the primitive RCC of the graft is variable. Partial nephrectomy is technically feasible and oncologically safe in the treatment of RCC of the renal graft. PMID- 19542594 TI - [CT SCAN as a predictor of composition and fragility of urinary lithiasis treated with extracorporeal shock wave lithotripsy in vitro]. AB - OBJECTIVES: To evaluate the ability of non contrast computed tomography (NCCT) to predict stone composition and fragility for treatment with extracorporeal shock wave lithotripsy (ESWL). METHODS: 27 stones of about 10 mm from patients who had undergone different endourological procedures were collected. All patients had been evaluated with NCCT. To perform in vitro ESWL an experimental device was designed. Three thousand pulses were applied with 17.2 Kv intensity using an electromagnetic generator (Lithostar) to all stones. Composition of each fragment was studied with crystallographic study. Results were statistically analyzed with Student Test, Chi2 Test and multivariate study. RESULTS: In vitro ESWL had a success rate of 59.26%. Average stone HU, grouped by composition: cistine 1015 HU, Calcium monohydrate oxalate 1193 HU, uric acid 419 HU, dihydrate calcium oxalate 2122 HU, struvite 1543 HU and basic phosphate magnesium 1517 HU. A statistically significant relationship was found between values which were lower than 500 HU and uric acid composition (p=0.0006), as well as values higher than 2000 HU and composition of dihydrated calcium acid (p=0.0244). In the group of stones with less than 1000 HU (n=11) efficacy was 81.1%, whereas it was 43.75% in the others (p=0.0479). We found a statistically significant relationship between uric acid and effectiveness (p=0.021). There was not statistically significant relationship between size and treatment effectiveness. CONCLUSIONS: The use of NCCT will allow predicting stone composition and fragility. PMID- 19542595 TI - [Testicular metastasis as first clinical expression of unknown origin malignant melanoma]. AB - OBJECTIVE: To report a rare process and to stimulate clinicians and pathologists to have this possibility present in their diagnoses. METHODS: We report the case of an adult male with pain and swelling in the left inguinal region and increased volume of the homolateral testicle. CONCLUSIONS: It is appropriate to bear in mind the possibility of testicular metastasis of a primary unknown tumor in adult males since metastatic affectation can constitute the first manifestation of the disease. PMID- 19542596 TI - [Current indications of open surgery for the treatment of renal lithiasis. Ureterocalycostomy as definitive treatment for lithiasis in a female with recurrent disease]. AB - OBJECTIVE: We describe one case of recurrent lithiasis associated with anatomical alteration of the renal pelvis related to previous surgery. METHODS/RESULTS: The patient presented a urinary tract infection episode, complicated with pyonephrosis and septicemia. In the intravenous urography, infectious radiopaque pyelocaliceal multiple and complex lithiasis can be seen, as well as kidney hydronephrosis grade III-IV. Important pyelic sclerosis secondary to previous surgery on the renal unit was seen. Nephrectomy was performed with lower pole nephro-lithotomy and reconstruction of the upper urinary tract through ureterocalicostomy. Two and a half years after surgery, control urogram shows absence of urolithiasis and a slight delay of renal function. CONCLUSIONS: Ureterocalicostomy is indicated in cases of ureteropelvic junction obstruction associated with intrarenal pelvis caused by alterations of fusion, rotation or location of kidney. It is also indicated in cases of severe peripyelic fibrosis secondary to previous pyeloplasty failure or renal surgery. In our case, in addition to the infectious component of lithiasis, an anatomical alteration, probably secondary to previous surgery, caused the chronification of lithiasis. Facing such suspicion a surgical management was undertaken to eliminate the lithiasis and get a correct derivation of the working area of the kidney, in order to prevent further recurrences. PMID- 19542597 TI - [Bilateral obstructive uropathy as clinical presentation of primary bladder lymphoma]. AB - OBJECTIVE: We report the event of an atypical presentation of primary bladder lymphoma, treated in our hospital, and review the literature of such tumors, representing 0.2% of bladder tumors, being macroscopic hematuria with clots the most frequent reason for patient consultation. METHODS: We report the case of an 83 years old man who went to the emergency room because of oligoanuria of 48 hours of evolution. He was diagnosed of bladder tumor. RESULTS: The pathology turned out to be a primary bladder lymphoma cell B. CONCLUSIONS: Primary bladder lymphoma is a rare entity that presents a similar behaviour to other bladder tumors, having a good response to treatment with chemotherapy. PMID- 19542598 TI - Assessment of visual field dependence: comparison between the mechanical 3D rod and-frame test developed by Oltman in 1968 with a 2D computer-based version. AB - The identification of subject's perceptual style regarding multisensory integration is a central issue for spatial perception and sensorimotricity. In spatial orientation studies, the weighting of visual frame of reference (visual field dependence) is classically assessed by using verticality perception tasks, and especially the mechanical 3D rod-and-frame test (3D RFT). The validation of a 2D computer-based version of the RFT by virtue of its portability would facilitate the identification of modes of spatial referencing for the design and evaluation of sensory and motor rehabilitation programs. We question here whether the computerized 2D RFT yields frame effects similar (in amplitude, direction) and correlated to those induced by the mechanical 3D RFT. In both devices, 35 young and healthy males' subjects were seated and tasked with aligning a rod to the gravity vertical within a square frame that was tilted at 18 degrees . The results showed significantly larger rod deviations from the verticality in the 3D RFT. 3D and 2D RFT errors significantly correlated but shared a small amount of common variance (r;{2} = 0.35). In addition, left-right tilt asymmetry changes from one device to another. These results suggest that the mechanical 3D RFT for verticality perception remains a more robust test for identifying the subject's perceptual style. PMID- 19542600 TI - Self-motion perception during conflicting visual-vestibular acceleration. AB - Self-motion is known to be falsely perceived during exposure to the movement of visual surroundings. This illusory perception of visually-induced self-motion is known as "vection." The present study was conducted to examine the relative strengths of vection versus whole-body angular acceleration as they determine perceived self-rotation under conditions in which they individually provide conflicting information. Each subject was rotated for 90 s about a vertical axis at a constant acceleration, and a large-field visual surround in front of the subject was simultaneously rotated at a constant acceleration in the same direction, but at a magnitude of acceleration twice that of the body. This stimulus condition creates a sensory conflict between information from the vestibular/somatosensory systems and information from the visual system with respect to the direction of self-rotation. The subject eventually perceived self acceleration in the direction of circular vection (CV), even though he or she was actually being accelerated in the direction opposite to CV. When the magnitude of contradictory chair acceleration exceeded the vestibular perceptual threshold, the onset latency of CV was significantly delayed. Our results suggest that visual information contributes to the perception of self-acceleration, and that illusory self-motion could overwhelm the feeling of self-acceleration due to inertial motion. CV would thus be a significant factor in determining spatial orientation in certain operational environments and flight conditions. PMID- 19542599 TI - Constructive perception of self-motion. AB - This review focusses attention on a ragged edge of our knowledge of self-motion perception, where understanding ends but there are experimental results to indicate that present approaches to analysis are inadequate. Although self-motion perception displays processes of "top-down" construction, it is typically analyzed as if it is nothing more than a deformation of the stimulus, using a "bottom-up" and input/output approach beginning with the transduction of the stimulus. Analysis often focusses on the extent to which passive transduction of the movement stimulus is accurate. Some perceptual processes that deform or transform the stimulus arise from the way known properties of sensory receptors contribute to perceptual accuracy or inaccuracy. However, further constructive processes in self-motion perception that involve discrete transformations are not well understood. We introduce constructive perception with a linguistic example which displays familiar discrete properties, then look closely at self-motion perception. Examples of self-motion perception begin with cases in which constructive processes transform particular properties of the stimulus. These transformations allow the nervous system to compose whole percepts of movement; that is, self-motion perception acts at a whole-movement level of analysis, rather than passively transducing individual cues. These whole-movement percepts may be paradoxical. In addition, a single stimulus may give rise to multiple perceptions. After reviewing self-motion perception studies, we discuss research methods for delineating principles of the constructed perception of self-motion. The habit of viewing self-motion illusions only as continuous deformations of the stimulus may be blinding the field to other perceptual phenomena, including those best characterized using the mathematics of discrete transformations or mathematical relationships relating sensory modalities in novel, sometimes discrete ways. Analysis of experiments such as these is required to mathematically formalize elements of self-motion perception, the transformations they may undergo, consistency principles, and logical structure underlying multiplicity of perceptions. Such analysis will lead to perceptual rules analogous to those recognized in visual perception. PMID- 19542601 TI - Effects of multi-directional vibrotactile feedback on vestibular-deficient postural performance during continuous multi-directional support surface perturbations. AB - Single-axis vibrotactile feedback of trunk tilt provided in real-time has previously been shown to significantly reduce the root-mean-square (RMS) trunk sway in subjects with vestibular loss during single-axis perturbation. This research examines the effect of multi-directional vibrotactile feedback on postural sway during continuous multi-directional surface perturbations when the subjects' eyes are closed. Eight subjects with vestibular loss donned a multi axis feedback device that mapped body tilt estimates onto their torsos with a 3 row by 16-column array of tactile actuators (tactors). Tactor row indicated tilt magnitude and tactor column indicated tilt direction. Root-mean-square trunk tilt, elliptical fits to trunk sway trajectory areas, percentage of time spent outside a no vibrotactile feedback zone, RMS center of pressure, and anchoring index parameters indicating intersegmental coordination were used to assess the efficacy of the multi-directional vibrotactile balance aid. Four tactor display configurations in addition to the tactors off configuration were evaluated. Subjects had significantly reduced RMS trunk sway, significantly smaller elliptical fits of the trajectory area, and spent significantly less time outside of the no feedback zone in the tactors on versus the tactors off configuration. Among the displays evaluated in this study, there was not an optimal tactor column configuration for standing tasks involving continuous surface perturbations. Furthermore, subjects performed worse when erroneous information was displayed. Therefore, a spatial resolution of 90 degrees (4 columns) seems to be as effective as a spatial resolution of 22.5 degrees (16 columns) for control of standing. PMID- 19542602 TI - Vibrotactile stimulators and virtual 3-D audio countermeasures, training and motion sickness symptoms with a simulated graveyard spin illusion. AB - The vestibular system by itself is incapable of effectively compensating for the graveyard spin illusion. We examined two countermeasures, i.e., efficacy of vibrotactile stimulation around the waist and virtual 3-D audio presented independently and jointly for controlling a simulated graveyard spin. We also examined: a) additional training with these countermeasures to improve "intuitiveness;" b) included non-perturbation trials along with perturbation trials; and c) monitored changes in well-being as measured by a motion sickness scale from pre- to post-test and immediately following each trial. Ten volunteers received two training and two test sessions. The somatogyral illusion was generated by accelerating a chair for 24 s until it attained a peak rotation of 120 degrees /s and then stopped. Over the ensuing 40 s the chair rotated in one of two random perturbation patterns or remained stationary. During this period, participants were required to eliminate all movements of the chair by turning a knob in the opposite direction of perceived rotation. For the control trials, participants relied solely on vestibular signals to cancel perceived movements. For the experimental trials, vibrotactile stimulation around the waist, virtual 3 D audio, or both were presented. The mean Cancellation Error (CE) for the control trials was 52 degrees /s. The application of the countermeasures significantly reduced the CE. Additional training and testing did not improve intuitiveness. Perturbations to the chair resulted in a higher CE than no perturbations. Motion sickness symptoms showed no differences from pre- to post-test and very rarely reported after each trial. PMID- 19542603 TI - Usefulness of some current balance tests for identifying individuals with disequilibrium due to vestibular impairments. AB - The goal of this study was to determine which of several clinical balance tests best identifies patients with vestibular disorders. We compared the scores of normals and patients on the Berg Balance Scale (Berg), Dynamic Gait Index (DGI), Timed Up and Go (TUG), Computerized Dynamic Posturography Sensory Organization Test (SOT), and a new obstacle avoidance test: the Functional Mobility Test (FMT). The study was performed in an out-patient balance laboratory at a tertiary care center. Subjects were 40 normal adults, and 40 adults with vestibular impairments. The main outcome measures were the sensitivity of tests to patients and specificity to normals. When adjusted for age the Berg, TUG, DGI and FMT had moderate sensitivity and specificity. SOT had moderately high sensitivity and specificity. SOT and FMT, combined, had high sensitivity and moderate specificity. Therefore, the kinds of tests of standing and walking balance that clinicians may use to screen patients for falling are not as good for screening for vestibular disorders as SOT. SOT combined with FMT is better. When screening patients for vestibular disorders, when objective diagnostic tests of the vestibular system, itself, are unavailable, tests of both standing and walking balance, together, give the most information about community-dwelling patients. These tests may also indicate the presence of sub-clinical balance problems in community-dwelling, asymptomatic adults. PMID- 19542604 TI - The microtubule-associated protein tau is also phosphorylated on tyrosine. AB - Tau protein is the principal component of the neurofibrillary tangles found in Alzheimer's disease (AD), where it is hyperphosphorylated on serine and threonine residues. It is hypothesized that this hyperphosphorylation contributes to neurodegeneration through the destabilization of microtubules. There is now evidence that phosphorylation of tau can also occur on tyrosine residues. Human tau has five tyrosines numbered 18, 29, 197, 310, and 394, according to the sequence of the longest CNS isoform. Tyrosines 18, 197, and 394 have been shown to be phosphorylated in the brain of patients with AD whereas tyrosine 394 is the only residue that has been described to date that is phosphorylated in physiological conditions. Src family kinases and spleen tyrosine kinase (Syk) have been shown to phosphorylate tyrosine 18 while c-Abl is capable of phosphorylating tyrosine 394. Recently, a dual specificity kinase termed TTBK1 has been characterized in human brain and shown to be able to phosphorylate residue 197 of tau. Data about the role of tau tyrosine phosphorylation in neuronal physiology are still scarce and preliminary. In contrast, there is mounting evidence suggesting that tau tyrosine phosphorylation is an early event in the pathophysiology of AD and that Fyn and c-Abl are critical in the neurodegenerative process which occurs in tauopathies. PMID- 19542605 TI - The role of neuropsychology in distinguishing the posterior cortical atrophy syndrome and Alzheimer's disease. AB - This study investigated the neuropsychological hallmarks of posterior cortical atrophy (PCA). Seventeen patients with PCA, 17 patients with probable Alzheimer's disease (PAD), and 17 healthy age-matched subjects underwent neuropsychological testing for abstract reasoning, visuospatial abilities, memory, language, executive functions, praxes, and attention. The PCA patients were significantly more impaired in visual perception, spatial memory, visual attention, and visuospatial reasoning compared to the PAD patients who were relatively more impaired in episodic memory. In the PCA group, no test score correlated with disease duration or age of clinical onset, whereas, in the PAD group, several scores correlated with disease duration. Compared to the healthy subjects, both patient groups showed multiple cognitive deficits. Thus, PCA is characterised by distinctive visuospatial deficits that reflect the distribution of brain damage and contrast with the memory impairment of PAD patients. Specific neuropsychological tests may contribute to early identification of cortical dementia for diagnostic and research purposes. PMID- 19542606 TI - Education attenuates the effect of medial temporal lobe atrophy on cognitive function in Alzheimer's disease: the MIRAGE study. AB - Functional imaging and neuropathological studies suggest that individuals with higher education have better cognitive performance at the same level of brain pathology than less educated subjects. No in vivo studies are available that directly test how education modifies the effect of structural pathology on cognition in Alzheimer's disease (AD). The present study therefore aimed to measure this effect using data from a large multi-center study. 270 patients with AD underwent cognitive testing using the Mini Mental State Examination (MMSE), apolipoprotein E (APOE) genotyping, and cerebral magnetic resonance imaging. A linear regression analysis was used to examine the relation of medial temporal lobe atrophy (MTA), as a proxy of AD pathology, to MMSE score, adjusting for age, gender, APOE, cerebrovascular disease, ethnicity, education, and disease duration. An interaction term for MTA and education was introduced to test the hypothesis that education modifies the effect of MTA on cognition. There was a significant inverse association between MTA and cognition. Most interestingly, the interaction term between education and MTA was significant suggesting that education modifies the relation of MTA to cognition. At any level of pathology, cognition remained higher for better educated individuals. PMID- 19542607 TI - High fruit and vegetable intake is positively correlated with antioxidant status and cognitive performance in healthy subjects. AB - A higher daily intake of fruits and vegetables in healthy elderly is associated with an improved antioxidant status in comparison to subjects consuming diets poor in fruits and vegetables, but the impact on cognitive performance is unclear. Healthy community dwellers (45 to 102 years old, n=193) underwent cognitive testing and blood withdrawal for the measurement of antioxidant micronutrients and biomarkers of oxidative stress as well as administration of a food frequency questionnaire to assess the daily intake of fruits and vegetables (high intake HI, low intake LI). Ninety-four subjects of the HI group had significantly higher cognitive test scores, higher levels of carotenoids, alpha- and gamma-tocopherol as well as lower levels of F2 alpha isoprostanes than the 99 subjects of the LI group. Cognitive scores were directly correlated with blood levels of alpha-tocopherol and lycopene and negatively correlated with F2 alpha isoprostanes and protein carbonyls. The results were independent of age, gender, body mass index, education, total cholesterol, LDL- and HDL-cholesterol, triglycerides, and albumin. Healthy subjects of any age with a high daily intake of fruits and vegetables have higher antioxidant levels, lower levels of biomarkers of oxidative stress, and better cognitive performance than healthy subjects of any age consuming low amounts of fruits and vegetables. Modification of nutritional habits aimed at increasing intake of fruits and vegetables should be encouraged to lower prevalence of cognitive impairment in later life. PMID- 19542608 TI - Alzheimer's amyloid-beta rescues yeast from hydroxide toxicity. AB - Amyloid-beta(Abeta42), which is known to be toxic to neuronal cells, protects yeast cells from severe sodium hydroxide toxicity. More than 85% cell death was caused by treatment with 1 mM NaOH and approximately 95% was observed at a 2 mM concentration. However, greater than 55% cells survived the treatment in the presence of Abeta42. A strong protective effect of the peptide was also evident from the differential staining of the treated culture with propidium iodide. PMID- 19542610 TI - Interactions between glycogen synthase kinase 3beta, protein kinase B, and protein phosphatase 2A in tau phosphorylation in mouse N2a neuroblastoma cells. AB - In this study, we investigated how tau phosphorylation is regulated by protein kinase glycogen synthase kinase 3beta (GSK3 beta), protein kinase B (PKB), and protein phosphatase 2A (PP2A) in mouse N2a neuroblastoma cells. Results showed that GSK3 beta overexpression significantly increased PKB phosphorylation at the S473 site but not the T308 site. Neither GSK3 beta nor PKB overexpression could reduce the PP2AC phosphorylation at the Y307 site. In contrast, either PKB or GSK3 beta knockdown could increase PP2A phosphorylation at the Y307 site. PP2AC knockdown increased GSK3 beta phosphorylation at the S9 site but not at the Y216 site, and PKB phosphorylation at the T308 site but not at the S473 site. Tau phosphorylation at the S396 site was increased by GSK3 beta or PKB overexpression. Tau phosphorylation at the S214 site was only induced by PKB overexpression in the study. While GSK3 beta knockdown decreased tau phosphorylation at the S396 site, PKB knockdown increased tau phosphorylation at both the S396 and S214 sites. PP2AC knockdown decreased tau phosphorylation at the S396 and S214 sites. These findings suggest that tau phosphorylation at the S396 and S214 sites is differentially regulated by GSK3 beta, PKB, and PP2A in N2a cells. The final phosphorylation state of tau is possibly caused by the synergic action of the three enzymes. PMID- 19542611 TI - On the diagnosis of CADASIL. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a genetic arteriopathy related to Notch3 mutations, is difficult to diagnosis. The goal of this study was to determine the value of clinical, immunohistochemical, and molecular techniques for the diagnosis of CADASIL. Clinical features and the immunohistochemical and molecular findings in 200 subjects with suspected CADASIL in whom 93 biopsies and 190 molecular studies are reported. Eighteen pathogenic mutations of the Notch3 gene, six of them previously unreported, were detected in 67 patients. The clinical features did not permit differentiation between CADASIL and CADASIL-like syndromes. The sensitivity and specificity of the skin biopsies was 97.7% and 56.5%, respectively, but increased to 100% and 81.5%, respectively, in cases with proven family history. In conclusion, a clinical diagnosis of CADASIL is difficult to determine and confirmatory techniques should be used judiciously. PMID- 19542612 TI - Temporal relationship between depressive symptoms and cognitive impairment: the Italian Longitudinal Study on Aging. AB - The temporal relationship between depression and cognitive decline has not been extensively investigated in prospective population-based studies, and most of these have only looked in one direction. We estimated the bidirectional temporal relationship between depressive symptoms and cognitive function in older subjects, excluding subjects with a clinical diagnosis of dementia or mild cognitive impairment (MCI). In a total of 2,963 individuals from the Italian Longitudinal Study on Aging, depressive symptoms, global cognitive function, and episodic memory were measured. Dementia, Alzheimer's disease, vascular dementia, and MCI were classified using current clinical criteria. Depressive symptoms at baseline were associated with an accelerated global cognitive function decline and an accelerated rate of episodic memory delayed recall decline in a 3.5-year follow-up. Finally, an accelerated increase with time of depressive symptoms during the same follow-up period was not associated with global cognitive function and episodic memory (immediate and delayed recall). In older subjects non-cognitively impaired, depressive symptoms at baseline predicted change over time of global cognitive decline and episodic memory delayed recall. Global cognitive function and episodic memory at baseline were not associated with the course of depressive symptoms during the follow-up. PMID- 19542614 TI - Total prion protein levels in the cerebrospinal fluid are reduced in patients with various neurological disorders. AB - We performed a study on levels of the total prion protein (PrP) in humans affected by different neurological diseases and assessed the influence of several factors such as age, gender, and disease severity on the cerebrospinal fluid PrP levels. PrP-ELISA technique was used to analyze cerebrospinal fluid (CSF) samples. 293 CSF samples of patients with Creutzfeldt-Jakob-disease (CJD), Alzheimer's disease, dementia with Lewy-bodies, Parkinson's disease, multiple sclerosis, cerebral ischemia, generalized epileptic seizures, and meningitis and encephalitis in comparison to controls were analyzed. We found a significant reduction of CSF PrP levels in patients suffering from all neurodegenerative disorders analyzed. This group exhibited mean PrP values of 164 ng/ml while non neurodegenerative disorder patients and healthy controls showed PrP levels of 208 ng/ml and 226 ng/ml, respectively. CSF levels correlated with disease severity in CJD, Alzheimer's disease, and dementia with Lewy-bodies. The finding of decreased PrP levels in the CSF of patients not only with CJD but also in other neurodegenerative disorders is intriguing. Age-, gender-, and genetic-specific factors might be involved in the PrP c regulation. PMID- 19542613 TI - BDNF variants, premorbid educational attainment, and disease characteristics in Alzheimer's disease: an exploratory study. AB - Brain-derived neurotrophic factor (BDNF) is a neurotrophin that promotes neuronal survival, growth, and differentiation. The role of BDNF in learning and memory suggests that it may also modulate the clinical course of Alzheimer's disease (AD). This study aimed to determine whether BDNF genetic variants are related to premorbid educational attainment, progression of cognitive and functional decline, and associated neuropsychiatric symptoms in AD patients. A sample of AD subjects (N = 341) was genotyped for the BDNF polymorphisms: Val66Met, C270T, and G-712A. Subjects received tests of cognition and daily function at baseline and at multiple subsequent time points. They were also characterized for the frequency and severity of neuropsychiatric symptoms. There was a significant effect of Val66Met genotype on educational attainment (F = 7.49, df = 2,329, P = 0.00066), with Met/Met homozygotes having significantly lower education than both the Val/Met and Val/Val groups. No association was observed between any BDNF polymorphism and measures of cognitive or functional decline. The T-allele of the C270T polymorphism was associated with a higher prevalence of neuropsychiatric symptoms and specifically with the presence of hallucinations. The effect of the Val66Met polymorphism on premorbid educational attainment is intriguing and should be verified in a larger sample. PMID- 19542615 TI - Neuropathology-based risk scoring for dementia diagnosis in the elderly. AB - Current neuropathologic consensus criteria for diagnosis of dementia yield a classification of processes that likely contributed to dementia in that individual. While dementia diagnosis currently relies on clinical criteria, practicing neuropathologists and researchers might benefit from a simple, accurate risk scoring protocol for the neuropathologic diagnosis of dementia. Using 232 consecutive autopsies from the population-based Adult Changes in Thought study, we developed two logistic regression-based risk scoring systems; one solely using neuropathologic measures and a second additionally including demographic information. Inverse-probability weighting was used to adjust for inherent selection bias in autopsy-based studies of dementing illnesses. Both systems displayed high levels of predictive accuracy; bias-adjusted area-under the-curve statistics were 0.78 (95% CI 0.71, 0.85) and 0.87 (95% CI 0.83, 0.92), indicating improved performance with the inclusion of demographic characteristics, specifically age and birth cohort information. Application of the combined neuropathology/demographic model yielded bias-adjusted sensitivity and specificity of 81% each. In contrast, application of NIA-Reagan criteria yielded sensitivity and specificity of 53% and 84%. Our proposed scoring systems provide neuropathologists with tools to make a diagnosis, and interpret their diagnosis in the light of known sensitivity and specificity estimates. Evaluation in independent samples will be important to verify our findings. PMID- 19542617 TI - Early changes in hippocampal Eph receptors precede the onset of memory decline in mouse models of Alzheimer's disease. AB - Synapse loss occurs early in Alzheimer's disease (AD) and is considered the best pathological correlate of cognitive decline. Ephrins and Eph receptors are involved in regulation of excitatory neurotransmission and play a role in cytoskeleton remodeling. We asked whether alterations in Eph receptors could underlie cognitive impairment in an AD mouse model overexpressing human amyloid beta protein precursor (hA beta PP) with familial mutations (hA beta PP swe-ind mice). We found that EphA4 and EphB2 receptors were reduced in the hippocampus before the development of impaired object recognition and spatial memory. Similar results were obtained in another line of transgenic A beta PP mice, Tg2576. A reduction in Eph receptor levels was also found in postmortem hippocampal tissue from patients with incipient AD. At the time of onset of memory decline inhA beta PP swe-ind mice, no change in surface expression of AMPA or NMDA receptor subunits was apparent, but we found changes in Eph-receptor downstream signaling, in particular a decrease in membrane-associated phosho-cofilin levels that may cause cytoskeletal changes and disrupted synaptic activity. Consistent with this finding, Eph receptor activation in cell culture increased phosho-cofilin levels. The results suggest that alterations in Eph receptors may play a role in synaptic dysfunction in the hippocampus leading to cognitive impairment in a model of AD. PMID- 19542618 TI - Vascular endothelial barrier dysfunction mediated by amyloid-beta proteins. AB - Neuronal inflammation is very common in Alzheimer's disease (AD). This inflammation can be caused by infiltration of neutrophils across the blood brain barrier. Endothelial permeability changes are required for the infiltration of high molecular weight components to the brain. Deposition of toxic amyloid-beta (A beta) fibrils in the cerebral vasculature, as well as in brain neurons, has been implicated in the development of AD. This study investigates the effect of A beta fibrils on the permeability of the endothelium and the mechanism for the observed permeability changes. A beta(1-40) and A beta(1-42) fibrils, but not monomers, were found to increase permeability of bovine pulmonary arterial endothelial cells in a dose- and time dependent manner as detected by transendothelial electrical resistance. This increase in permeability is only partially (25%) inhibited by catalase and is not associated with an increase in cytosolic Ca+2 or tyrosine phosphorylation. These results indicate that hydrogen peroxide is not the primary mediator for the permeability changes. Treatment of cells with both amyloid fibrils resulted in stress fiber formation, disruption and aggregation of actin filaments, and cellular gap formation. The results of this study reveal that A beta increases the permeability of endothelium by inducing change in the cytoskeleton network. PMID- 19542619 TI - Expression of a truncated human tau protein induces aqueous-phase free radicals in a rat model of tauopathy: implications for targeted antioxidative therapy. AB - Oxidative stress has been implicated in the pathogenesis of many neurodegenerative diseases including Alzheimer's disease (AD). We investigated the effect of a truncated form of the human tau protein in the neurons of transgenic rats. Using electron paramagnetic resonance we observed significantly increased accumulation of ascorbyl free radicals in brains of transgenic animals (up to 1.5-fold increase; P < 0.01). Examination of an in vitro model of cultured rat corticohippocampal neurons revealed that even relatively low level expression of human truncated tau protein (equal to 50% of endogenous tau) induced oxidative stress that resulted in increased depolarization of mitochondria (approximately 1.2-fold above control, P < 0.01) and increases in reactive oxygen species (approximately 1.3-fold above control, P < 0.001). We show that mitochondrial damage-associated oxidative stress is an early event in neurodegeneration. Furthermore, using two common antioxidants (vitamin C and E), we were able significantly eliminate tau-induced elevation of reactive oxygen species. Interestingly, vitamin C was found to be selective in the scavenging activity, suggesting that expression of truncated tau protein preferentially leads to increases in aqueous phase oxidants and free radicals such as hydrogen peroxide and hydroxyl and superoxide radicals. Our results suggest that antioxidant strategies designed to treat AD should focus on elimination of aqueous phase oxidants and free radicals. PMID- 19542616 TI - The neurodegenerative mitochondriopathies. AB - Mitochondria are physically or functionally altered in many neurodegenerative diseases. This is the case for very rare neurodegenerative disorders as well as extremely common age-related ones such as Alzheimer's disease and Parkinson's disease. In some disorders very specific patterns of altered mitochondrial function or systemic mitochondrial dysfunction are demonstrable. Some disorders arise from mitochondrial DNA mutation, some from nuclear gene mutation, and for some the etiology is not definitively known. This review classifies neurodegenerative diseases using mitochondrial dysfunction as a unifying feature, and in doing so defines a group of disorders called the neurodegenerative mitochondriopathies. It discusses what mitochondrial abnormalities have been identified in various neurodegenerative diseases, what is currently known about the mitochondria-neurodegeneration nexus, and speculates on the significance of mitochondrial function in some disorders not classically thought of as mitochondriopathies. PMID- 19542620 TI - The accuracy of Medicare claims as an epidemiological tool: the case of dementia revisited. AB - Our study estimates the sensitivity and specificity of Medicare claims to identify clinically-diagnosed dementia, and documents how errors in dementia assessment affect dementia cost estimates. We compared Medicare claims from 1993 2005 to clinical dementia assessments carried out in 2001-2003 for the Aging Demographics and Memory Study (ADAMS) cohort (n = 758) of the Health and Retirement Study. The sensitivity and specificity of Medicare claims was 0.85 and 0.89 for dementia (0.64 and 0.95 for AD). Persons with dementia cost the Medicare program (in 2003) $7,135 more than controls (P < 0.001) when using claims to identify dementia, compared to $5,684 more when using ADAMS (P < 0.001). Using Medicare claims to identify dementia results in a 110% increase in costs for those with dementia as compared to a 68% increase when using ADAMS to identify disease, net of other variables. Persons with false positive Medicare claims notations of dementia were the most expensive group of subjects ($11,294 versus $4,065, for true negatives P < 0.001). Medicare claims overcount the true prevalence of dementia, but there are both false positive and negative assessments of disease. The use of Medicare claims to identify dementia results in an overstatement of the increase in Medicare costs that are due to dementia. PMID- 19542622 TI - Albumin-blunted deleterious effect of amyloid-beta by preventing the internalization of the peptide into neurons. AB - Amyloid-beta (A beta) is the main component of senile plaques, one of the hallmarks of Alzheimer's disease. Our results showed that A beta(25-35) decreased neuronal viability while it increased generation of reactive oxygen species (ROS). Under these circumstances, albumin (BSA) prevented ROS production and neuronal death in a dose- and time-dependent manner. In addition, BSA partially prevented the decrease in the expression of GAP-43, MAP-2, and tubulin, and the phosphorylation of tau protein caused by A beta, suggesting that BSA protects against the loss of plasticity caused by the peptide. Our findings suggest that BSA exerts its protective effect by binding to A beta in an equimolecular way, which prevents heterodimer (A beta-BSA) entry into neurons. In fact, BSA prevented A beta internalization, as shown by confocal immunocytochemistry, suggesting that BSA causes its protective effect by sequestrating A beta, which cannot reach its intracellular targets. This is consistent with the idea that A beta must enter neurons to exert its deleterious effects. PMID- 19542624 TI - Immunoglobulin GM genes as functional risk and protective factors for the development of Alzheimer's disease. AB - There is growing body of evidence for the involvement of herpes simplex virus type 1 (HSV1) in the etiology of Alzheimer's disease (AD). HSV1 has evolved strategies for decreasing the efficacy of the host immune response and interfering with viral clearance. Based on their putative role as the modulators of these immune avoidance strategies, I hypothesize that immunoglobulin (Ig) GM genes - genetic markers of IgG heavy chains located on chromosome 14 - are functional risk and protective factors for AD. Results from genome-wide association and linkage studies in support of this hypothesis, testable predictions, and possible therapeutic implications are discussed. PMID- 19542623 TI - SPECT predictors of cognitive decline and Alzheimer's disease in mild cognitive impairment. AB - Baseline brain single photon emission computed tomography (SPECT) was evaluated in eighty subjects with mild cognitive impairment (MCI) who were followed for a mean of about two years, when twelve patients developed Alzheimer's disease (AD), nineteen showed memory decline (D), and forty-three had normal cognition assessment (stable: S) (six drop-out). Volumetric Regions of Interest (VROI) analysis was performed in six associative cortical areas in each hemisphere. ANOVA for repeated measures showed significant effects for both the group (S, D, and AD; p < 0.004) and VROI (p < 0.0001) factors, with significant group*region interaction (p < 0.01). At post-hoc comparison, hippocampal VROIs values were lower in AD than in D and S, while parietal VROIs values were lower in D and AD than in S. These four VROI significantly correlated with verbal delayed recall score at follow-up visit. Receiver operating characteristic (ROC) curves for the mean hippocampal VROI value showed 0.81 sensitivity with 0.86 specificity in separation of S+D from AD (p < 0.0001), and 0.69 sensitivity with 0.75 specificity in separation of S from D+AD (p < 0.0002). ROC curves for the mean parietal VROI value showed 0.62 sensitivity with 0.70 specificity in separation of S from D+AD (p < 0.0002). Baseline SPECT can support outcome prediction in subjects with MCI. PMID- 19542625 TI - Why did tarenflurbil fail in Alzheimer's disease? AB - There has been a lot of disappointment surrounding the recent failure of the largest ever study in patients with Alzheimer's disease (AD) with tarenflurbil, a compound believed to modulate the activity of gamma-secretase, the pivotal enzyme that generates the amyloid-beta (A beta) peptide from the amyloid-beta protein precursor. What are the reasons for this setback after the previous apparently encouraging results in a Phase II study? A straightforward explanation of this failure is that the gamma-secretase is not the right target for therapy or that, in general, blocking A beta does not produce clinical benefits in AD. If one still accepts a physiopathological role of A beta in AD, tarenflurbil could not be the right compound because of its weak pharmacological activity as an A beta(1 42) lowering agent and its poor brain penetration. In addition, based on previous negative results with several anti-inflammatory drugs in AD, it is hypothesized that the residual anti-inflammatory activity of tarenflurbil may have a detrimental effect on disease progression. PMID- 19542621 TI - Mechanisms of nitrosamine-mediated neurodegeneration: potential relevance to sporadic Alzheimer's disease. AB - Streptozotocin (STZ) is a nitrosamine-related compound that causes Alzheimer's disease (AD)-type neurodegeneration with cognitive impairment, brain insulin resistance, and brain insulin deficiency. Nitrosamines and STZ mediate their adverse effects by causing DNA damage, oxidative stress, lipid peroxidation, pro inflammatory cytokine activation, and cell death, all of which occur in AD. We tested the hypothesis that exposure to N-nitrosodiethylamine (NDEA), which is widely present in processed/preserved foods, causes AD-type molecular and biochemical abnormalities in central nervous system (CNS) neurons. NDEA treatment of cultured post-mitotic rat CNS neurons (48 h) produced dose-dependent impairments in ATP production and mitochondrial function, and increased levels of 8-hydroxy-2'-deoxyguanosine, 4-hydroxy-2-nonenal, phospho-tau, amyloid-beta protein precursor-amyloid-beta (A beta PP-A beta), and ubiquitin immunoreactivity. These effects were associated with decreased expression of insulin, insulin-like growth factor (IGF)-I, and IGF-II receptors, and choline acetyltransferase. Nitrosamine exposure causes neurodegeneration with a number of molecular and biochemical features of AD including impairments in energy metabolism, insulin/IGF signaling mechanisms, and acetylcholine homeostasis, together with increased levels of oxidative stress, DNA damage, and A beta PP-A beta immunoreactivity. These results suggest that environmental exposures and food contaminants may play critical roles in the pathogenesis of sporadic AD. PMID- 19542626 TI - Amyloid-beta, tau, and dementia. AB - Alzheimer's disease (AD) is clinically characterized as a progressive dementia starting with memory dysfunction and characterized pathologically as neurodegeneration accompanied by deposition of amyloid-beta, neurofibrillary tangles, and neuronal loss. AD research has endeavored to explain the clinical symptoms of AD through pathological changes and to develop various therapies for AD. Fulfillment of these goals, however, remains on the horizon. In this article, I review the relationship between neuropathological changes that occur in the brain and clinical progression of AD, and propose a hypothesis that brain aging, characterized by neurofibrillary tangles in entorhinal cortex, is pre-requisite for development of AD. PMID- 19542627 TI - A systematic review of neuropsychiatric symptoms in mild cognitive impairment. AB - Mild cognitive impairment (MCI) is a clinical concept proposed as an intermediate state between normal aging and dementia. This condition has multiple heterogeneous sources, including clinical presentation, etiology, and prognosis. Recently, the prevalence and associated features of neuropsychiatric symptoms (NPS) in MCI have been described. We systematically searched the PubMed database (last accessed on August 31, 2008) for articles on NPS in MCI. Included articles used strict selection criteria, and outcome variables were extracted in duplicate; of the 27 articles included, 14 (52%) used prospective cohorts. The global prevalence of NPS in MCI ranged from 35% to 85%. The most common behavioral symptoms were depression, anxiety, and irritability. Hospital-based samples reported a higher global prevalence of NPS than population-based studies; this discrepancy probably reflected differences in demographics, study setting, MCI diagnostic criteria, and behavioral instruments used. Prospective studies showed that NPS, particularly depression, may represent risk factors for MCI or predictors for the conversion of MCI to Alzheimer's disease (AD). NPS are very prevalent in subjects with MCI, displaying a similar pattern of symptoms compared to dementia and AD. Large cohort studies using standardized MCI criteria and behavioral instruments are required to evaluate the prognostic role of NPS in MCI. PMID- 19542628 TI - Validity of cerebrospinal fluid biomarkers as endpoints in early-phase clinical trials for Alzheimer's disease. AB - A systematic literature review was performed to assess the suitability of cerebrospinal fluid (CSF) levels of protein amyloid-beta42 (Abeta42) and tau as markers to detect the disease-modifying effects of drugs in clinical trials of AD treatments. All databases were searched for observational studies, single-arm clinical trials, and randomized controlled trials involving patients with AD in which CSF Abeta42 and tau were measured. A meta-analytic random-effects model was used to evaluate the mean absolute change in protein concentration over time. Spearman correlation was used to assess the association between change in CSF protein concentration and change in cognitive function. The mean changes per month in observational studies were -0.4 pg/ml/month (95% CI: -1.9 to 1.1) for 8 CSF Abeta42 and 1.5 pg/ml/month (95% CI: 0.1 to 3.0) for 12 CSF tau studies. The correlation coefficients for the relationship between CSF protein concentration and cognition were 0.43 (p = 0.068) for all 18 Abeta42 studies and -0.05 (p = 0.857) for all 18 tau studies. A trend in which CSF Abeta42 decreases and tau protein increases over time was identified in AD patients. CSF Abeta42 and tau concentrations should be used with caution as surrogate endpoints in early-phase clinical trials for AD. PMID- 19542629 TI - Systemic inflammation and the risk of Alzheimer's disease and dementia: a prospective population-based study. AB - Inflammation is suggested to be involved in the pathogenesis of Alzheimer's disease (AD). Serum interleukin-6 (IL-6) and high sensitivity serum reactive protein C (hsCRP) as markers of systemic inflammation were analyzed at two examinations of the ULSAM-study, a longitudinal, community-based study of elderly men (age 70, n = 1062 and age 77, n = 749). In addition, serum amyloid protein A (SAA) and urinary prostaglandin F2alpha (PGF2alpha) metabolite levels were analyzed at age 77 in this cohort. Two serial samples (at ages 70 and 77) were available from 704 individuals. Using Cox regression analyses, associations between serum IL-6, hsCRP, SAA and PGF2alpha metabolite levels and risk of AD, any type of dementia (all-cause dementia) and non-AD dementia were analyzed. On follow-up (median, 11.3 years) in the age 70 cohort, 81 subjects developed AD and 165 subjects developed all-cause dementia. Serum IL-6, hsCRP, SAA, or PGF2alpha levels were not associated with risk of AD. At age 70, high IL-6 levels were associated with an increased risk of non-AD dementia (Hazard ratio 2.21 for above vs. below/at median, 95%confidence interval 1.23-3.95, p-value = 0.008). A longitudinal change in CRP or IL-6 levels was not associated with AD ordementia. In conclusion, Serum IL-6, hsCRP, SAA, and PGF2alpha levels are not associated with the risk of AD. High serum IL-6 levels may be associated with increased risk of non-AD dementia. PMID- 19542630 TI - Insulin is a two-edged knife on the brain. AB - Insulin, long known as an important regulator of blood glucose levels, plays important and multifaceted roles in the brain. It has been reported that insulin is an important neuromodulator, contributing to several neurobiological processes in particular energy homeostasis and cognition. Dysregulation of insulin signaling has been linked to aging and metabolic and neurodegenerative disorders. The first part of this review is devoted to discussion of the critical role of insulin signaling in normal brain function. Then the involvement of impaired insulin signaling in the pathophysiology of diabetes, Alzheimer's, Parkinson's and Huntington's diseases and amyotrophic lateral sclerosis will be discussed. Finally, the potential therapeutic effect of insulin and insulin sensitizers will be examined. PMID- 19542632 TI - Systemic tocopherols and F2-isoprostanes and the risk of Alzheimer's disease and dementia: a prospective population-based study. AB - Oxidative stress in the brain is suggested to be involved in the pathophysiology of Alzheimer's disease (AD). In this study, serum alpha- and gamma-tocopherol, the two major systemic antioxidants, were analyzed at two examinations of the ULSAM-study, a longitudinal, community-based study of elderly men (age 70, n = 616 and age 77, n = 761). In addition, urinary F2-isoprostane levels, as markers of systemic oxidative stress, were analyzed at the age of 77 in this cohort (n = 679). Cox regression analyses were used to examine associations between serum alpha-, gamma-tocopherol and urinary F2-isoprostane levels and AD, any type of dementia (all-cause dementia) and non-AD dementia. On follow-up (median, 12.3 years), 40 subjects developed AD and 86 subjects developed all-cause dementia. Serum alpha- and gamma-tocopherol or urinary F2-isoprostane levels were not associated with the future risk of AD or dementia. In conclusion, systemic serum alpha- and gamma-tocopherol and urinary F2-isoprostane levels are not associated with the future risk of AD or dementia and do not seem to be useful predictors of clinical AD or dementia. PMID- 19542633 TI - Coffee "breaks" Alzheimer's disease. PMID- 19542634 TI - Antibiotic resistant bacteria in wastewater processed by the Metropolitan Water Reclamation District of Greater Chicago system. AB - The Metropolitan Water Reclamation District of Greater Chicago (District) initiated a research study to determine the total numbers and percentages of antibiotic resistant fecal coliform (FC) bacteria in raw sewage (RS) entering and final effluents (FE) discharged from its seven Water Reclamation Plants (WRPs). The density of FC was determined on m-FC agar containing ampicillin (ampR-16 microg/ml), gentamycin (genR-8 microg/ml), tetracycline (tetR-8 microg/ml), or all three antibiotics. The study was primarily undertaken to determine whether secondary sewage treatment at the District WRPs adequately reduces the numbers and percentages of FC(ampR), FC(tetR), FC(genR), FC(amp/tet/genR) in the FE. The numbers of ampR, tetR, genR, and amp/tet/genR FC observed in RS ranged from 2.0 x 10(5) to 1.1 x 10(7), 9.5 x 10(4) to 2.2 x 10(6), 95 to 1.5 x 10(4) and 90 to 9.5 x 10(3) per 100 mL, respectively. Secondary sewage treatment without disinfection was shown to reduce the number of antibiotic resistant FC by two-three orders of magnitude. The numbers of FC(ampR), FC(tetR), FC(genR), and FC(amp/tet/genR) observed in non-disinfected FE ranged from 2.0 x 10(2) to 6.4 x 10(3), 2.2 x 10(2) to 4.1 x 10(3), 9 to <20 and 9 to <20 per 100 mL, respectively. The relative percentages of antibiotic resistant FC observed in FE followed the same trend observed in RS: FC(ampR) > FC(tetR) > FC(genR) > FC(amp/tet/genR). Only one FC(amp/tet/genR) bacteria was found in this study indicating that multiple antibiotic resistant FC was virtually eliminated by secondary sewage treatment. The results of multivariate regression analysis showed that the percentages of antibiotic resistant FC in the FE from all seven District WRPs were lower than the percentages of these organisms in RS (p<0.01). These results support the conclusion that secondary sewage treatment in the District effectively reduces the number of antibiotic resistant FC and that the environments of the District's seven WRPs are not conducive to the propagation or survival of antibiotic resistant fecal coliform bacteria. PMID- 19542631 TI - Mapping cofilin-actin rods in stressed hippocampal slices and the role of cdc42 in amyloid-beta-induced rods. AB - Dissociated hippocampal neurons exposed to a variety of degenerative stimuli form neuritic cofilin-actin rods. Here we report on stimulus driven regional rod formation in organotypic hippocampal slices. Ultrastructural analysis of rods formed in slices demonstrates mitochondria and vesicles become entrapped within some rods. We developed a template for combining and mapping data from multiple slices, enabling statistical analysis for the identification of vulnerable sub regions. Amyloid-beta (Abeta) induces rods predominantly in the dentate gyrus region, and Abeta-induced rods are reversible following washout. Rods that persist 24 h following transient (30 min) ATP-depletion are broadly distributed, whereas rods formed in response to excitotoxic glutamate localize within and nearby the pyramidal neurons. Time-lapse imaging of cofilin-GFP-expressing neurons within slices shows neuronal rod formation begins rapidly and peaks by 10 min of anoxia. In approximately 50% of responding neurons, Abeta-induced rod formation acts via cdc42, an upstream regulator of cofilin. These new observations support a role for cofilin-actin rods in stress-induced disruption of cargo transport and synaptic function within hippocampal neurons and suggest both cdc42-dependent and independent pathways modulate cofilin activity downstream from Abeta. PMID- 19542635 TI - Stochastic approach to the derivation of emission limits for wastewater treatment plants. AB - Stochastic approach to the derivation of WWTP emission limits meeting probabilistically defined environmental quality standards (EQS) is presented. The stochastic model is based on the mixing equation with input data defined by probability density distributions and solved by Monte Carlo simulations. The approach was tested on a study catchment for total phosphorus (P(tot)). The model assumes input variables independency which was proved for the dry-weather situation. Discharges and P(tot) concentrations both in the study creek and WWTP effluent follow log-normal probability distribution. Variation coefficients of P(tot) concentrations differ considerably along the stream (c(v)=0.415-0.884). The selected value of the variation coefficient (c(v)=0.420) affects the derived mean value (C(mean)=0.13 mg/l) of the P(tot) EQS (C(90)=0.2 mg/l). Even after supposed improvement of water quality upstream of the WWTP to the level of the P(tot) EQS, the WWTP emission limits calculated would be lower than the values of the best available technology (BAT). Thus, minimum dilution ratios for the meaningful application of the combined approach to the derivation of P(tot) emission limits for Czech streams are discussed. PMID- 19542636 TI - Two-phase flow modelling for oxygen renewal estimation in vertical flow filter: luxury or necessity? AB - Scientists and practitioners exhibit an increasing interest on effluent transfer and degradation modelling in Vertical Flow Sand Filters (VFSF) and Vertical Flow Constructed Wetland (VFCW). Modelling software used to this purpose is mainly monophasic: in the unsaturated zone, only water flow is taken into account and air phase influence is assumed to be negligible. In hydrology, many studies have point out the limitations of this assumption in order to quantify air phase movement but little has been done in the modelling of vertical flow filter. Despite its complexity, two-phase flow modelling allows to overcome these difficulties. In this work, we describe the complex air and water flows in the particular case of vertical flow filter fed intermittently using both numerical and experimental results. Complete different behaviour is observed depending on ponding occurs or not. If it does, flow is clearly influenced by air entrapment which is responsible of a reduction of the infiltration speed and of the drainage of a part of the water kept at the interface between the sand and the drainage layer. Finally, we study the dependency of oxygen income by convection on hydraulic load and compare numerical results with experimental results obtained on oxygen consumption. PMID- 19542638 TI - Stochastic long term modelling of a drainage system with estimation of return period uncertainty. AB - Long term prediction of maximum water levels and combined sewer overflow (CSO) in drainage systems are associated with large uncertainties. Especially on rainfall inputs, parameters, and assessment of return periods. This paper proposes a Monte Carlo based methodology for stochastic prediction of both maximum water levels as well as CSO volumes based on operations of the urban drainage model MOUSE in a single catchment case study. Results show quite a wide confidence interval of the model predictions especially on the large return periods. Traditionally, return periods of drainage system predictions are based on ranking, but this paper proposes a new methodology for the assessment of return periods. Based on statistics of characteristic rainfall parameters and correlation with drainage system predictions, it is possible to predict return periods more reliably, and with smaller confidence bands compared to the traditional methodology. PMID- 19542637 TI - Water quality and occurrence of water-borne diseases in the Douala 4th District, Cameroon. AB - The monthly occurrence and mean age distribution of water-borne diseases in the Douala 4th District, Cameroon (1995-2006) were studied and probable causes of diseases spread were established. Diseases of interest included gastroenteritis, amoebic dysentery, typhoid fever and cholera. Water-borne disease occurrence was observed to follow a seasonal pattern with peaks occurring between the months of January and May followed by drops between June and October and rose again from November. Children below 5 years were found to be more vulnerable to diarrhoea, gastroenteritis, amoebic dysentery while persons between 15-44 years were more vulnerable to typhoid and cholera. Physico-chemically, water samples had turbidities varying between 5.5-86 NTU, pH values between 4.2 and 7.1 and zero residual chlorine. Bacteriological analysis showed that the total coliform count was averagely 74/100 ml, the faecal colform count was 43/100 ml and the faecal streptococci count was 27/100 ml. Lack of access to potable water, absence of sanitation facilities and environmental factors could be advanced as the probable causes of water-borne disease spread. PMID- 19542639 TI - Community-focused greywater management in two informal settlements in South Africa. AB - South Africa is struggling to provide services to the millions of poor people migrating to the major centres and living in informal settlements (shanty towns). Whilst the local authorities are generally able to provide potable water from the municipal network to communal taps scattered around the settlements, there is usually inadequate provision of sanitation and little or no provision for the drainage of either stormwater or greywater. This paper describes an investigation into ways of engaging with community structures in the settlements with a view to encouraging "self-help" solutions to greywater management requiring minimal capital investment as an interim "crisis" solution until such time that local and national government is able to provide formal services to everyone. The work was carried out in three settlements encompassing a range of different conditions. Only two are described here. It has become clear that the management of greywater has a low priority amongst the residents of informal settlements. The lack of effective political structures and the breakdown in communication with Ward Councillors and local government officials have contributed to the lack of progress. As the project progressed it became evident that greywater cannot be considered separately from stormwater, sanitation and refuse removal. PMID- 19542640 TI - Acute toxicity and relationship between metabolites and ecotoxicity during the biodegradation process of non-ionic surfactants: fatty-alcohol ethoxylates, nonylphenol polyethoxylate and alkylpolyglucosides. AB - The toxicity values of fatty-alcohol ethoxylates, nonylphenol polyethoxylate, and alkylpolyglucosides have been determined by applying assays with luminescent bacteria. Also, the relation between metabolites and ecotoxicity during the biodegradation process has been determined. The biodegradation tests were carried out according to the OECD 301 E test for ready biodegradability. In these tests a solution of the surfactant, representing the sole carbon source for the microorganisms, was tested in a mineral medium, inoculated and incubated under aerobic conditions in the dark. The toxicity of surfactants is related to their molecular structure (Quantitative Structure Activity Relationships, QSAR). For the alkylpolyglucosides, toxicity expressed as EC(50) is related with the critical micelle concentration (CMC), the hydrophilic-lipophilic balance (HLB) of the surfactant, and the hydrophobic alkyl chain (R). The results indicate that toxicity increased as the CMC decreased and as the hydrophobicity increased and R rose. For fatty-alcohol ethoxylates, parameters characteristic studied have been HLB, number of units of ethylene oxide and the alkyl chain length. Relationships found are in agreement with the fact that increasing the alkyl chain length leads to a lower EC(50), whereas increasing ethoxylation leads to a lower toxicity. An analysis of the behaviour of the toxicity and HLB again indicates that the toxicity was greater for surfactants with a smaller HLB. The evolution of the toxicity was studied over the biodegradation process, expressed as a percentage of inhibition. For all the non-ionic surfactants assayed, except for the nonylphenol polyethoxylate, a major decline was found in toxicity during the first days of the biodegradation assay and at all the concentrations tested. PMID- 19542641 TI - Pollutant removal from oily wastewater discharged from car washes through sedimentation-coagulation. AB - Wastewater from car washes represents a potential problem for the sewer system due to its emulsified oils and suspended material. Treatment of wastewater discharged from four car washes was investigated by sedimentation and coagulation. The effect of the coagulants Servical P (aluminium hydroxychloride), Servican 50 (poly(diallyldimethylammonium chloride)), aluminium sulfate and ferric chloride was evaluated. The achieved removal using sedimentation was of 82%, 88% 73% and 51% for oils, total suspended solids, COD, and turbidity, respectively. In the treatment by coagulation we achieved average efficiencies nearly to 74% for COD removal, greater than 88% in the case of total suspended solids removal and 92% in the case of turbidity and except the performance of Servican 50 greater than 90% in oil removal. We concluded that the oil residual concentration and COD in the treated water allows pouring it in the sewer system complying with the limits of the Mexican rule NOM-002-ECOL-1996 and it is possible even its reuse, at least in the case of the chassis washing of cars. PMID- 19542642 TI - Advanced nitrogen removal via nitrite from municipal wastewater in a pilot-plant sequencing batch reactor. AB - To obtain economically sustainable wastewater treatment, advanced nitrogen removal from municipal wastewater and the feasibility of achieving and stabilizing short-cut nitrification and denitrification were investigated in a pilot-plant sequencing batch reactor (SBR) with a working volume of 54 m(3). Advanced nitrogen removal, from summer to winter, with effluent TN lower than 3 mg/L and nitrogen removal efficiency above 98% was successfully achieved in pulsed-feed SBR. Through long-term application of process control in pulsed-feed SBR, nitrite accumulation reached above 95% at normal temperature of 25 degrees C. Even in winter, at the lowest temperature of 13 degrees C, nitrite was still the end production of nitrification and nitrite accumulation was higher than 90%. On the basis of achieving advanced nitrogen removal, short-cut nitrification and denitrification was also successfully achieved. Compare to the pulse-feed SBR with fixed time control, the dosage of carbon source and energy consumption in pulsed-feed SBR with process control were saved about 30% and 15% respectively. In pulsed-feed SBR with process control, nitrogen removal efficiency was greatly improved. Moreover, consumption of power and carbon source was further saved. PMID- 19542643 TI - Nitrogen removal performance of anaerobic ammonia oxidation co-culture immobilized in different gel carriers. AB - Four materials were prepared as carriers for immobilizing anaerobic ammonium oxidizing sludge. Nitrogen removal performance by these immobilized gel bead groups was evaluated. The removal ratios of ammonium and nitrite by CMC anammox immobilized beads were 100% and 95.3% in 48 hours, respectively. The removal efficiencies of ammonium and nitrite by SA, PVA-SA and PVA anammox-immobilized beads were lower than the CMC beads. Subsequently, the physical properties of the beads were studied. PVA-SA was found to be the best support material among the four by comparing the case of the immobilization procedure, nitrogen removal efficiencies, and the costs of materials. PVA-SA gel entrapment was optimized by an orthogonal experiment. The SEM micrographs displayed that the surface structure of PVA-SA immobilized beads is loose and finely porous, which facilitates diffusion of the nitrogen. The SEM micrographs also clearly showed that anammox bacteria existed in the gel beads. All results clearly demonstrate that immobilizing anammox sludge in gel carriers is feasible and exhibit good performance. This research provided a new route to maintain sufficient amount of anammox sludge in a practical anammox reactor. PMID- 19542644 TI - Preparation of activated carbons from wet activated sludge by direct chemical activation. AB - An improved method for preparing activated carbons from wet waste activated sludge (WAS) by direct chemical activation was studied in this paper. The effects of processing parameters on iodine adsorption capacity of the product were investigated. Results show that sludge-based activated carbon prepared with KOH had a larger iodine value than those activated with ZnCl(2) and KCl. The maximum iodine value was observed at the KOH concentration of 0.50 M. Increasing the impregnation time from 10 to 20 h resulted in a 20% increase in the iodine value. The highest iodine value was obtained at the activation temperature of 600 degrees C and holding time of 1 h. Sludge water content had insignificant effects on the iodine value of products. Raw WAS with a water content of 93.2% can be converted into an activated carbon with a high specific surface area of 737.6 m(2) g(-1) and iodine value of 864.8 mgg(-1) under optimum experimental conditions. Other physical properties such as total pore volume, micropore volume and mean pore diameter of the product were also reported and compared with those of commercial activated carbon. PMID- 19542645 TI - Developing a multipoint titration method with a variable dose implementation for anaerobic digestion monitoring. AB - Determination of metabolites from an anaerobic digester with an acid base titration is considered as superior method for many reasons. This paper describes a practical at line compatible multipoint titration method. The titration procedure was improved by speed and data quality. A simple and novel control algorithm for estimating a variable titrant dose was derived for this purpose. This non-linear PI-controller like algorithm does not require any preliminary information from sample. Performance of this controller is superior compared to traditional linear PI-controllers. In addition, simplification for presenting polyprotic acids as a sum of multiple monoprotic acids is introduced along with a mathematical error examination. A method for inclusion of the ionic strength effect with stepwise iteration is shown. The titration model is presented with matrix notations enabling simple computation of all concentration estimates. All methods and algorithms are illustrated in the experimental part. A linear correlation better than 0.999 was obtained for both acetate and phosphate used as model compounds with slopes of 0.98 and 1.00 and average standard deviations of 0.6% and 0.8%, respectively. Furthermore, insensitivity of the presented method for overlapping buffer capacity curves was shown. PMID- 19542646 TI - Indole degrading of ammonia oxidizing bacteria isolated from swine wastewater treatment system. AB - Three new strains named LPA11, LPB11 and LPC24 were isolated to investigate the patterns of indole degradation and ammonia oxidation in swine wastewater from different parts of a swine wastewater treatment system by the direct spreading plate method. These three isolates were all identified as Pseudomonas putida based on 16S-rDNA gene sequences, main physiological and biochemical analysis. They were capable of decomposing 1.0 mM indole completely in 10, 16 and 18 days respectively. According to the results of HPLC and GC/MS, the possible pathway for the degradation was via oxindole, isatin and anthranilic acid. The three bacteria were capable of oxidizing ammonia, and the strains LPA11 and LPC24 were capable of effectively reducing nitrate and nitrite. PMID- 19542647 TI - Upflow anaerobic clarification tank (UACT) to upgrade existing anaerobic effluents. AB - The Upflow Anaerobic Clarification Tank (UACT) is of interest as a post-treatment step for high-rate anaerobic wastewater treatment systems. The UACT contributes to extra capture of methane from the raw wastewater. It also significantly facilitates the reuse of the treated water. The efficiency of the UACT process is demonstrated in this study using laboratory-scale reactors. The water produced by the UACT had a low total COD, in most cases below 145 mg/L and a soluble COD below 60 mg/L at surface loading rates between 0.1-0.4 m(3)/(m(2) h) and hydraulic retention times between 5-6 h. The sludge bed of the UACT was characterised by 6-10 times higher substrate affinity (for ethanol, acetate and propionate) compared to the parent UASB and similar specific activity. The results suggest that a UASB followed by the UACT can, upon subsequent application of a rapid filtration, qualify as a total COD removal, no longer necessitating aerobic treatment. PMID- 19542648 TI - Evaluation of reflection and refraction in simulations of ultraviolet disinfection using the discrete ordinates radiation model. AB - Simulations of UV disinfection systems require accurate models of UV radiation within the reactor. Processes such as reflection and refraction at surfaces within the reactor can impact the intensity of the simulated radiation field, which in turn impacts the simulated dose and performance of the UV reactor. This paper describes a detailed discrete ordinates radiation model and comparisons to a test that recorded the UV radiation distribution around a low pressure UV lamp in a water-filled chamber with a UV transmittance of 88%. The effects of reflection and refraction at the quartz sleeve were investigated, along with the impact of wall reflection from the interior surfaces of the chamber. Results showed that the inclusion of wall reflection improved matches between predicted and measured values of incident radiation throughout the chamber. The difference between simulations with and without reflection ranged from several percent near the lamp to nearly 40% further away from the lamp. Neglecting reflection and refraction at the quartz sleeve increased the simulated radiation near the lamp and reduced the simulated radiation further away from the lamp. However, the distribution and trends in the simulated radiation field both with and without the effects of reflection and refraction at the quartz sleeve were consistent with the measured data distributions. PMID- 19542649 TI - Simulation on combined rapid gravity filtration and backwash models. AB - Combined rapid gravity filtration and backwash models have been applied to simulate filtration and backwash cycles. The simulated results from the backwash model suggest that an optimum air flow rate exists to maximise particle removal efficiency in the backwash operation for a certain backwash system. The simulation of combined rapid gravity filtration and backwash models suggests that the filter should not be completely cleaned up in the backwash and a certain amount of particles retained on filter grains after backwash can be beneficial for subsequent filtration runs. This is consistent with the experimental results in the literature. PMID- 19542650 TI - Effect of environmental flow management on river water quality: a case study at Yeongsan River, Korea. AB - This paper describes a management scheme to control river water quality using additional water discharges from upstream dams, which results in an increase environmental flow (EF) followed by an enhancement of water quality in a target river. To suggest a creditable management plan among a suite of ideal scenarios, the monthly averaged water quality monitoring data from 2001 to 2006 at the Yeongsan (YS) River, Korea were investigated with respect to seasonal variation and spatial distribution. From the analysis, it was found that while biochemical oxygen demand (BOD(5)) level in the YS River was extremely high during the dry/drought season (April, May, and June; AMJ), the level was subsequently decreased during the monsoon season (July, August, and September; JAS) due mainly to the dilution effect of rainfall. To improve the water quality in AMJ, we here suggested a scenario of increasing EF using surplus water discharges from upstream dams, which was examined by one dimensional riverine water quality model, QUAL2E model. Simulation result showed that additional discharge from the upstream dams could lead, on average, to a 36% of water quality improvement in mainstream with respect to BOD(5). Model coefficients were validated by comparing the six year monitoring data to minimize a sum of squares error, and showed a good agreement with the observed data. Overall, the methodology developed in this paper appears to be quite clear and straightforward, and thus, can be applied to a wide range of the flow managements or water quality controls in a stream with artificial structures. PMID- 19542651 TI - Evaluation of effectiveness of chemical and physical sewage treatment technologies for removal of retinoic acid receptor agonistic activity detected in sewage effluent. AB - Retinoic acid receptor (RAR) is a nuclear receptor involved in vertebrate morphogenesis, growth, cellular differentiation, and tissue homeostasis. Excess expression of the retinoid signaling can cause various developmental toxicities in animals and humans. We previously found that influents from sewage treatment plants (STPs) in Japan had a RAR agonistic activity and the activity cannot be removed completely by conventional biological treatments. In this study, we assessed the performance of chemical and physical sewage treatment technologies ozonation, ultraviolet treatment, chlorination, coagulation using polyaluminium chloride (PAC) and ferric sulfate, and filtration with ultrafiltration (UF), nanofiltration (NF), and reverse osmosis (RO) membranes-in removal of RAR agonistic activity of STP effluent. All water treatment experiments were conducted in laboratory-scale reactors. The RAR agonistic activity of samples was measured using a yeast two-hybrid assay. Results showed that the effectiveness of tested technologies on the removal of RAR agonistic activity can be ranked as RO or NF > chlorination > ozonation > MF > UV > coagulation with ferric sulfate>>coagulation with PAC. Furthermore, the effectiveness of chlorination might rank lower because excess reaction might bring a side effect by producing some RAR agonistic by-product(s). PMID- 19542652 TI - Hydraulic performance of a mature wetland treating milkhouse wastewater and agricultural runoff. AB - A tracer study is an efficient method of determining flow dynamics within a constructed wetland. In previous studies, a number of tracer studies have been carried out on various constructed wetlands covering a wide range of configurations. From these tracer studies it is evident that all constructed wetlands perform differently and generally with less efficiency than assumed by theoretical design computations. During the summer of 2004, a tracer study was performed on a constructed wetland located in Embrun, Ontario (Canada) treating milkhouse wastewater and agricultural runoff to determine its actual hydraulic performance. Sediment height and vegetation density profiles were also obtained and examined to explain the preferential flow pathways that were observed during the tracer analysis. It was determined that the constructed wetland had an effective treatment area representing 79% of the total area, and that the hydraulic efficiency of the system was 74%. Examination of the sediment height and vegetation density profiles resulted in no evidence of physical pathways that could be attributed to the establishment of preferential flow. The hydraulic efficiency was therefore attributed to the inflow and outflow layout of the constructed wetland cell, combined with wind induced mixing. PMID- 19542653 TI - Characterisation of titanium tetrachloride and titanium sulfate flocculation in wastewater treatment. AB - Flocculation with titanium tetrachloride (TiCl(4)) and titanium sulfate (Ti(SO(4))(2)) was investigated in terms of different coagulant doses, pH, turbidity, dissolved organic carbon (DOC), UV-254, colour, zeta potential, particle size and molecular weight distribution. The two coagulants were compared with the commonly used coagulants such as ferric chloride (FeCl(3)) and aluminium sulfate (Al(2)(SO(4))(3)). Titanium tetrachloride showed the highest turbidity removal, while titanium sulfate showed the highest reduction of UV-254 and colour at all pH values. The four coagulants were found to have similar organic removal up to 60-67% and resulted in similar organic removal in terms of various MW ranges. The decantability of the settled flocs was very high for titanium tetrachloride, titanium sulfate and ferric chloride compared with aluminium sulfate. The dominating coagulation mechanisms for titanium tetrachloride and titanium sulfate are still to be studied, since different precipitation reactions might take place at different pH even without flocculant addition. Titanium tetrachloride and titanium sulfate were found as effective new coagulants in wastewater treatment not only in terms of organic matter removal, but also in sludge reduction through the production of titanium dioxide. PMID- 19542654 TI - Performance of electron beam irradiation for treatment of groundwater contaminated with acetone. AB - The purposes of this study were to evaluate the efficiency of acetone removal by electron beam irradiation in groundwater and the effect of various conditions. According to the results, the removal kinetics of acetone were pseudo first order, and the removal efficiencies were expressed to the (%) removal and G values. By adding sulfite, it was confirmed that acetone was mainly degraded by the reaction with the hydrated electrons. The presence of nitrate caused the removal of acetone to decrease. But there was no significant effect of alkalinity on the removal of acetone. The effect of the initial pH values (pH 5 to 9) on the acetone removal efficiency was negligible, but the pH value decreases due to the formation of acidic compounds after irradiation. Consequently, the radiation induced removal reactions of acetone followed the pseudo-first-order kinetic model; in addition to the initial concentration of acetone, nitrate and the absorbed dose were important factors in removing acetone from an aqueous solution using electron beam irradiation. The effects of general pH and alkalinity on the degrading acetone were negligible. PMID- 19542655 TI - Medium-term performance and maintenance of SUDS: a case-study of Hopwood Park Motorway Service Area, UK. AB - One of the main barriers to implementing SUDS is concern about performance and maintenance costs since there are few well-documented case-studies. This paper summarizes studies conducted between 2000 and 2008 of the performance and maintenance of four SUDS management trains constructed in 1999 at the Hopwood Park Motorway Service Area, central England. Assessments were made of the wildlife value and sedimentation in the SUDS ponds, the hydraulic performance of the coach park management train, water quality in all management trains, and soil/sediment composition in the grass filter strip, interceptor and ponds. Maintenance procedures and costs were also reviewed. Results demonstrate the benefits of a management train approach over individual SUDS units for flow attenuation, water treatment, spillage containment and maintenance. Peak flows, pond sediment depth and contaminant concentrations in sediment and water decreased through the coach park management train. Of the 2007 annual landscape budget of pounds 15,000 for the whole site, the maintenance costs for SUDS only accounted for pounds 2,500 compared to pounds 4,000 for conventional drainage structures. Furthermore, since sediment has been attenuated in the management trains, the cost of sediment removal after the recommended period of three years was only pounds 554 and, if the design is not compromised, less frequent removal will be required in future. PMID- 19542656 TI - Development of statistical linear regression model for metals from transportation land uses. AB - The transportation landuses possessing impervious surfaces such as highways, parking lots, roads, and bridges were recognized as the highly polluted non-point sources (NPSs) in the urban areas. Lots of pollutants from urban transportation are accumulating on the paved surfaces during dry periods and are washed-off during a storm. In Korea, the identification and monitoring of NPSs still represent a great challenge. Since 2004, the Ministry of Environment (MOE) has been engaged in several researches and monitoring to develop stormwater management policies and treatment systems for future implementation. The data over 131 storm events during May 2004 to September 2008 at eleven sites were analyzed to identify correlation relationships between particulates and metals, and to develop simple linear regression (SLR) model to estimate event mean concentration (EMC). Results indicate that there was no significant relationship between metals and TSS EMC. However, the SLR estimation models although not providing useful results are valuable indicators of high uncertainties that NPS pollution possess. Therefore, long term monitoring employing proper methods and precise statistical analysis of the data should be undertaken to eliminate these uncertainties. PMID- 19542657 TI - Fabrication of zero valent iron (ZVI) nanotube film via potentiostatic anodization and electroreduction. AB - Zero valent iron has been successfully used for the degradation of a wide range of contaminants. However, this reaction of using ZVI particle produces a large quantity of iron sludge. To solve the problem, we report the synthesis of self organized nanoporous zero valent iron film treated with anodization and electro reduction of iron foil. The iron nanotubes were fabricated in 1 M Na(2)SO(4) + 0.5 wt% NaF electrolyte by supplying constant electric currents of 50 mV/s, and holding the potential at 20, 40 and 60 V for 20 min. Nanoporous shape was produced by anodic oxidation of iron film. After anodizing process, electro reduction of nanoporous iron film converted crystallization iron oxide to zero valent iron. Electro-reduction process was carried out by electro-reducing with powersupply to and holding the potential at 20 V for 20 min. The surface of iron nanotube film was examined by BET and the thickness of the oxidized films was evaluated by scanning electron microscope (SEM). The crystalline structures of the fabricated films were evaluated using X-ray diffraction (XRD). PMID- 19542659 TI - Gaining from gaming. PMID- 19542658 TI - Effect of key operating parameters on the non-catalytic wet oxidation of olive mill wastewaters. AB - The non-catalytic wet air oxidation (WAO) of olive mill wastewaters was investigated. The effect of operating conditions, such as initial organic loading (1,000 and 4,500 mg/L COD), reaction temperature (140 and 180 degrees C), treatment time (1 and 4 h), initial pH (4.8 and 7) and the use of 500 mg/L H(2)O(2) as an additional oxidant, on treatment efficiency was assessed implementing a factorial experimental design. Of the five parameters tested, the first two had a considerable effect on COD removal, while treatment time was of no significance implying that all oxidation reactions occur during the first hour of treatment. Although the level of mineralization was generally moderate, this was accompanied by nearly complete total phenols and color removal. The analysis was repeated at more intense conditions, i.e. initial COD up to 8,000 mg/L and reaction temperature up to 200 degrees C; at this level, none of the studied effects were important. However, at optimal experimental conditions (i.e. 180 degrees C, 1 h treatment and initial COD of 8,100 mg/L) WAO yielded 34, 94 and 74% removal of COD, total phenols and color respectively. Moreover, ecotoxicity to V. fischeri was slightly reduced after 2 h of treatment at the above conditions. PMID- 19542660 TI - Phase 1 trials in oncology in India: the clear case for innovation. PMID- 19542661 TI - Role of human papilloma virus in the oral carcinogenesis: an Indian perspective. AB - Oral squamous cell carcinoma (OSCC) is one of the most common cancers in the Indian subcontinent. Although tobacco and alcohol are the main etiologic factors for nearly three-fourth of these cancers, no definite etiologic factor can be identified in one-fourth of the cases. There is growing evidence that human papilloma virus (HPV) may act as a cocarcinogen, along with tobacco, in the causation of oral cancers. The role of HPV in the etiology of anogenital cancers has been firmly established, and infection with this virus has also been shown to have prognostic significance. However, there is no clear evidence to support its involvement in oral carcinogenesis. We searched the PubMed database for all literature published from 1985 to 2008 and performed a systemic review in order to understand the relationship of HPV with oral cancers and its prevalence in various sub-sites in the oral cavity. Association of HPV is strongest for oropharyngeal cancers, especially cancers of the tonsils, followed by those of the base of tongue. High-risk HPV-16 is the predominant type; it commonly affects the younger age-groups, with males appearing to have a predisposition for infection with this strain. Its prevalence increases from normal to dysplasia and finally to cancer. HPV prevalence has been reported to be twice as high in premalignant lesions as in normal mucosa and is nearly five times higher in OSCC. The overall prevalence of HPV in OSCC ranges between 20-50%. OSCCs associated with HPV have been found to have better outcomes, being more responsive to radiotherapy and showing higher survival rates. In view of the association of HPV with OSCC, it should be worthwhile to conduct further experimental studies to elucidate its role in oral carcinogenesis. PMID- 19542662 TI - Normal tissue complication probability: does simultaneous integrated boost intensity-modulated radiotherapy score over other techniques in treatment of prostate adenocarcinoma. AB - AIM: The main objective of this study was to analyze the radiobiological effect of different treatment strategies on high-risk prostate adenocarcinoma. MATERIALS AND METHODS: Ten cases of high-risk prostate adenocarcinoma were selected for this dosimetric study. Four different treatment strategies used for treating prostate cancer were compared. Conventional four-field box technique covering prostate and nodal volumes followed by three-field conformal boost (3D + 3DCRT), four-field box technique followed by intensity-modulated radiotherapy (IMRT) boost (3D + IMRT), IMRT followed by IMRT boost (IMRT + IMRT), and simultaneous integrated boost IMRT (SIBIMRT) were compared in terms of tumor control probability (TCP) and normal tissue complication probability (NTCP). The dose prescription except for SIBIMRT was 45 Gy in 25 fractions for the prostate and nodal volumes in the initial phase and 27 Gy in 15 fractions for the prostate in the boost phase. For SIBIMRT, equivalent doses were calculated using biologically equivalent dose assuming the alpha/beta ratio of 1.5 Gy with a dose prescription of 60.75 Gy for the gross tumor volume (GTV) and 45 Gy for the clinical target volume in 25 fractions. IMRT plans were made with 15-MV equispaced seven coplanar fields. NTCP was calculated using the Lyman-Kutcher-Burman (LKB) model. RESULTS: An NTCP of 10.7 +/- 0.99%, 8.36 +/- 0.66%, 6.72 +/- 0.85%, and 1.45 +/- 0.11% for the bladder and 14.9 +/- 0.99%, 14.04 +/- 0.66%, 11.38 +/- 0.85%, 5.12 +/- 0.11% for the rectum was seen with 3D + 3DCRT, 3D + IMRT, IMRT + IMRT, and SIBIMRT respectively. CONCLUSIONS: SIBIMRT had the least NTCP over all other strategies with a reduced treatment time (3 weeks less). It should be the technique of choice for dose escalation in prostate carcinoma. PMID- 19542663 TI - Is electroglottography-based videostroboscopic assessment of post-laryngectomy prosthetic speech useful? AB - OBJECTIVES: To use an electroglottography (EGG)-based videostroboscopy tool to assess the anatomical and morphologic characteristics of the pharyngoesophageal (PE) segment in tracheoesophageal (TO) speakers. STUDY DESIGN: Cross-sectional cohort study. SUBJECTS: Fifty-two post-laryngectomy patients with no recurrence and using prosthetic (Blom-Singer) speech. INTERVENTION: An electroglottography (EGG)-based videostroboscopy tool EGG-based rigid videostroboscopy as well as perceptual evaluation. OUTCOME MEASURES: Stroboscopic protocol included nine subjective/visual parameters to evaluate the neoglottis and study correlation of the G (GRBAS scale) and the overall voice quality (OVQ) with the treatment variables. RESULTS: Of the 52 laryngectomees, videostroboscopic recordings were possible in 46 patients (36 males and 10 females) with a mean age of 63.4 +/-10.5 (SD) an electroglottography (EGG)-based videostroboscopy tool years. All used the Blom-Singer valve and the median time since Total Laryngectomy was 2 years. The neoglottis was assessable in 26 patients. We were able to strobe only 9 patients. There was excellent correlation between G and OVQ (Spearman rho > 0.9). Statistically significant correlation was found between G1 and saliva (P = 0.03) and between good OVQ and saliva (P = 0.02); similarly, there was significant correlation between G1 and LVV (P = 0.05) and between good OVQ and LVV (P = 0.03). CONCLUSIONS: This study is the first to examine the use of an EGG-based stroboscopy instrument to evaluate TO speech. Our observations suggest that from the standpoint of functional voice, saliva and the LVV had statistically significant effect in determining voice quality. PMID- 19542664 TI - Accelerated partial-breast irradiation vs conventional whole-breast radiotherapy in early breast cancer: a case-control study of disease control, cosmesis, and complications. AB - CONTEXT: Accelerated partial-breast irradiation (APBI) using various approaches is being increasingly employed for selected women with early breast cancer (EBC). AIMS: To conduct a case-control study comparing disease control, cosmesis, and complications in patients with EBC undergoing APBI using multicatheter interstitial brachytherapy vs those receiving conventional whole breast radiotherapy (WBRT). SETTINGS AND DESIGN: Women with EBC fulfilling the American Brachytherapy Society (ABS) criteria were selected as 'cases' if treated with APBI or as 'controls' if offered WBRT during the period from May 2000 to December 2004. MATERIALS AND METHODS: APBI patients were treated with high-dose-rate brachytherapy (HDR) to a dose of 34 Gy/10#/6-8 days. WBRT was delivered to the whole breast to a dose of 45 Gy/25# followed by tumor bed boost, either with electrons (15 Gy/6#) or interstitial brachytherapy (HDR 10 Gy/1#). RESULTS: At the median follow-up of 43.05 months in APBI and 51.08 months in WBRT there was no difference in overall survival (OS), disease-free survival (DFS), late arm edema, and symptomatic fat necrosis between the two groups. However, APBI resulted in increase in mild breast fibrosis at the tumor bed. Telangiectasias were observed in three patients of the APBI group. The cosmetic outcome was significantly better in the APBI group as compared to the WBRT group (P = 0.003). CONCLUSIONS: This study revealed equivalent locoregional and distant disease control in the two groups. APBI offered better overall cosmetic outcome, though at the cost of a slight increase in mild breast fibrosis and telangiectasias. PMID- 19542665 TI - Results of combined modality treatment for nasopharyngeal cancer. AB - CONTEXT: Radiotherapy is the cornerstone of treatment in nasopharyngeal cancer (NPC); the addition of chemotherapy has shown improved results. AIMS: To compare the results of concurrent chemoradiation with that of radiotherapy alone in NPC. MATERIALS AND METHODS: One hundred and ninety consecutive NPC patients, without distant metastasis, who reported to the institute from January 1992 to December 2001, received external-beam radiation to 66 Gy in 33 fractions. Seventy-five of these patients received concurrent chemotherapy with cisplatin and 5-fluorouracil (5-FU) for four cycles. We compared the results of treatment in these two groups. RESULTS: The 5-year disease-free survival rates were 40% and 60%, respectively, for patients who had radiotherapy alone and those who had chemoradiation (P = 0.002), while the median survival was 45 months and 60 months, respectively (P = 0.0028). CONCLUSION: A significant improvement in local control and survival was observed by the addition of concurrent chemotherapy with cisplatin and 5-FU to radical radiation in this nonrandomized study on patients with NPC. PMID- 19542667 TI - A simple technique for cranio-spinal irradiation in pediatric patients. AB - PURPOSE: Field matching poses challenges in craniospinal irradiation (CSI) as it leads either to underdosage or overdosage in the junctional area. A simple technique for CSI in pediatric patients is proposed. MATERIALS AND METHODS: Computed tomography scans were performed in the prone position. Two lateral cranial fields and a direct posterior spinal field were planned with a common central axis. Half-beam-blocked cranial fields with zero collimator rotation were used for treating the cranium. A half-beam-blocked field defined with jaws was used to treat the spinal column at an extended source-to-surface distance. Before treating the patient, matching of the cranial and spinal field junction along the central axis was verified with an extended dose range film. RESULTS AND CONCLUSION: The technique described is simple and easy to implement and can be applied to pediatric patients undergoing CSI. This method has the potential to reduce daily setup time and setup errors. This technique is ideally suitable for patients with spinal fields less than 30 cm. PMID- 19542666 TI - Time trial: a prospective comparative study of the time-resource burden for three dimensional conformal radiotherapy and intensity-modulated radiotherapy in head and neck cancers. AB - INTRODUCTION: An ongoing institutional randomized clinical trial comparing three dimensional conformal radiotherapy (3D CRT) and intensity-modulated radiotherapy (IMRT) provided us an opportunity to document and compare the time-manpower burden with these high-precision techniques in head and neck cancers. MATERIALS AND METHODS: A cohort of 20 consecutive patients in the ongoing trial was studied. The radiotherapy planning and delivery process was divided into well defined steps and allocated human resource based on prevalent departmental practice. Person-hours for each step were calculated. RESULTS: Twelve patients underwent IMRT and eight patients had 3D CRT. The prerandomization steps (upto and including approval of contours) were common between the two arms, and expectedly, the time taken to complete each step was similar. The planning step was carried out postrandomization and the median times were similar for 3D CRT (312 min, 5.2 person-hours) and IMRT (325.6 min, 5.4 person-hours). The median treatment delivery time taken per fraction varied between the two arms, with 3D CRT taking 15.2 min (0.6 person-hours), while IMRT taking 27.8 min (0.9 person hours) (P< 0.001). The total treatment time was also significantly longer in the IMRT arm (median 27.7 versus 17.8 person-hours, P< 0.001). The entire process of IMRT took 48.5 person-hours while 3D CRT took a median of 37.3 person-hours. The monitor units delivered per fraction and the actual "beam-on" time was also statistically longer with IMRT. CONCLUSIONS: IMRT required more person-hours than 3D CRT, the main difference being in the time taken to deliver the step-and-shoot IMRT and the patient-specific quality assurance associated with IMRT. PMID- 19542668 TI - Resource-sparing and cost-effective strategies in current management of breast cancer. AB - Breast cancer is the leading cause of death in women throughout the world. There have been significant advances in the practice of breast oncology over the past few years. However, most of these advances have an associated price tag or are resource intensive. The present article discusses means to achieve cost effectiveness in the treatment of breast cancer, while retaining the benefits of the modern anticancer approaches. PMID- 19542669 TI - The great debate: stroboscopy vs high- speed imaging for assessment of alaryngeal phonation. AB - Assessment of total laryngectomy patients has always been problematic and controversial. This is particularly so when one wants to assess and characterize the mucosal wave using the existing modalities of assessment. Videostroboscopy is generally regarded as an easily available and clinically relevant technique for adequately assessing this important parameter. However, it has its limitations, especially in laryngectomies. Recently, high-speed imaging has been recommended as the ideal tool for studying the physiology and mucosal parameters in this set of patients. In this brief article we examine this debate on the 'best' modality for assessment of the total laryngectomy patient. PMID- 19542670 TI - Cylindric cell carcinoma of the base of the tongue: a rare variant of squamous cell carcinoma. AB - Cylindric cell carcinomas (transitional cell carcinomas) are a rare and distinct histopathological entity presenting in the head and neck region. They have been known by myriads of nomenclature like cylindric carcinomas, nonkeratinizing sinonasal carcinoma, papillary carcinoma, cylindrical or columnar cell carcinoma, intermediate cell carcinoma, Schneiderian carcinoma, and Ringertz carcinoma. They are considered a variant of nonkeratinizing squamous cell carcinoma. Cylindric carcinomas are usually described in the sinus and nasal cavity and rarely said to involve nasopharynx and larynx. Only passing references have been made for its presentation in oropharynx including tonsils and the base of the tongue. We report here a rare case of transitional cell carcinoma presenting in the base of the tongue. There are no separate treatment recommendations in the literature, and the management is on the lines of treatment of squamous cell carcinoma. We report here a case of cylindric cell carcinoma presenting in the base of the tongue. The patient was staged as having cT2 N3 M0 (Stage IV B) disease. The patient received palliative radiotherapy of 20 Gy in five fractions followed by chemotherapy with injection paclitaxel and carboplatin. A partial response to treatment was achieved at the time of writing this report. PMID- 19542671 TI - Sister Mary Joseph nodule as the presenting sign of disseminated prostate carcinoma. AB - Sister Mary Joseph's nodule is referred to as metastasis of visceral malignancy to the umbilicus. Most common primaries are in the gastrointestinal or genital tract, while other locations are rare. We recently encountered a 76-year-old male who was referred to the surgery clinic with an erythematous nodule in the umbilicus measuring 6 cm in diameter with complaints of painless profuse hematuria. History revealed severe obstructive voiding symptoms of 2-year duration, along with significant loss of weight and difficulty in walking. A detailed examination showed hard nodular hepatomegaly, along with grade IV prostatomegaly. Serum prostate-specific antigen was 3069 ng/ml. A pelvic radiograph displayed multiple osteolytic lesions, while ultrasonography showed multiple iso- and hypoechoic lesions in both lobes of the liver, suggestive of metastasis. Histopathology of a Tru-cut biopsy of the prostate confirmed an adenocarcinoma (Gleason score 9) with umbilical metastasis. The patient was on regular follow-up and died 3 months later. PMID- 19542672 TI - Dural sinus vein thrombosis in a patient with colon cancer treated with FOLFIRI/bevacizumab. AB - The adverse effects of regimes in cancer treatment have forced us to change to new targeted therapy options. Understanding these side effects, which can lead to discontinuation of the new therapy strategies, will allow the clinical management of these side effects and result in continuing therapies with effective medications. Bevacizumab, which is an IgG1 antibody against vascular endothelial growth factor, has side effects such as proteinuria, hypertension, venous and arterial thromboembolic events, and hemorrhage. This is the first reported case of dural sinus vein thrombosis, during the treatment with bevacizumab. PMID- 19542674 TI - Brain metastasis from esophageal carcinoma. AB - Esophageal carcinoma rarely metastasizes to brain. In our center, among 504 cases of esophageal cancer registered for treatment during a 15-year (1990-2005) period, brain metastasis from esophageal carcinoma was detected in only 1 case. An unusual case of esophageal carcinoma that presented with brain metastasis is reported here. PMID- 19542673 TI - Small cell carcinoma of the urinary bladder: a case report and review of the literature. AB - Small cell carcinoma of the bladder (SCCB) is extremely rare. In this paper, we present a case of metastatic SCCB managed by chemotherapy and we would provide a brief review of the epidemiology, clinical features, diagnosis, pathologic features, staging, treatment, and prognosis of SCCB. A 52-year-old man was admitted with signs and symptoms suggestive of a bladder cancer. Computed tomography of the pelvis and abdomen showed a large tumor at the right bladder wall, measuring 10 cm in diameter, and a multinodular liver disease. Diagnosis of small cell carcinoma was established from the histological study of the transurethral resection of the bladder tumor. The patient received 12 cycles of platinum-based chemotherapy with a good partial response of bladder tumor and liver metastasis. The patient is still alive, 18 months after diagnosis. PMID- 19542675 TI - Diffuse large B-cell lymphoma of the uterine cervix: a rare case managed novelly. AB - Non-Hodgkin's lymphoma (NHL) of the uterine cervix is exceedingly rare. The management of the disease is not standardized. A 44-year-old lady presented with a history of bleeding pervaginum and a foul-smelling discharge of 2 months' duration. A 7 x 7 growth was seen in the cervix. A biopsy revealed it to be a CD20-positive diffuse large B-cell (DLBCL)-type NHL. She was diagnosed as stage IE after staging work-up, and managed with three courses of rituximab, cyclophosphamide, vincristine, adriamycin, and prednisolone followed by external beam radiotherapy (46 Gy in 23 fractions) by 3D conformal technique. She attained a complete response, and has been in remission for 1 year 3 months. PMID- 19542676 TI - Higher prevalence of dry symptoms in skin, eyes, nose and throat among workers in clean rooms with moderate humidity. AB - OBJECTIVE: To determine whether working under relative humidity (RH) around 55 +/ 5% may lead to dry symptoms among workers in tropical regions. METHODS: We recruited 3,154 Taiwanese workers who had no history of skin diseases and compared dry symptoms between clean room workers (RH around 55 +/- 5%) and other workers (RH around 65 +/- 5%). RESULTS: Clean room workers had higher prevalences of dry symptoms of the eye (odds ratio [OR]=1.62, 95% confidence interval [CI]: 1.40 to 1.86), nose and throat (OR=2.15, 95% CI: 1.66 to 2.79), and skin (OR=1.46, 95% CI: 1.23 to 1.73). In clean room workers, however, dry skin symptoms affected the palms (OR=1.72, 95% CI: 1.24 to 2.39), which are covered by gloves, more frequently than the face (OR=0.65, 95% CI: 0.45 to 0.94), which is exposed to the room air. We found working in clean rooms (adjusted OR [AOR]=1.38, 95% CI: 1.08 to 1.77), 24 to 30 yr of age (AOR=0.78, 95% CI: 0.62 to 0.99), family history of atopic diseases (AOR=1.75, 95% CI: 1.37 to 2.25), and skin moisturizer use (AOR=1.64, 95% CI: 1.30 to 2.06) were independent predictors of skin symptoms. In addition, working in clean rooms was an independent predictor of dry eye (AOR=1.30, 95% CI: 1.06 to 1.60) and dry nose and throat (AOR=1.70, 95% CI: 1.28 to 2.26) symptoms. CONCLUSIONS: Whereas the humidity in such working environments is not very low, for workers living in a high humidity environment, the relatively low humidity may still cause dry symptoms of the eye, nose, and throat. PMID- 19542677 TI - Participatory action oriented training for hospital nurses (PAOTHN) program to prevent musculoskeletal disorders. AB - OBJECTIVES: The purpose of this study was to construct and test the feasibility and potential utility of a Participatory Action Oriented Training for Hospital Nurses (PAOTHN) program to prevent work-related musculoskeletal disorders. The PAOTHN program emphasized the active participation of nurses, resulting in practical and low-cost solutions for improving their work environment by reducing risk factors of musculoskeletal disorders. METHODS: The PAOTHN program was conducted in a hospital located in a suburb of Seoul, South Korea. Of the 24 units in the hospital, 16 units participated in the study. The main components of the intervention were a series of structured workshops, continuous technical assistance by the research team, and periodical forums for sharing "best practices" among the participants. RESULTS: Through the workshops, head nurses of the participating units identified a total of 46 strategic (23 short-term and 23 long-term) plans for reducing musculoskeletal disorders risks across five dimensions: (1) patient care and treatment, (2) safe handling of drugs, medical devices, and equipment, (3) workstation design, (4) physical environment, and (5) welfare facilities and administration. Over the course of the year-long project, 18 of the suggested plans were completed, for an overall completion rate of 39.1%. CONCLUSIONS: The PAOTHN program was found to be feasible and potentially useful in reducing the musculoskeletal disorder risks faced by hospital nurses, and in identifying both risk factors and improvement opportunities at the individual and organizational levels. PMID- 19542678 TI - Optimization of the determination of ortho-phthalaldehyde in air by derivatization with 2,4-dinitorophenylhyrazine (DNPH). PMID- 19542679 TI - Rapid and effective speciation analysis of arsenic compounds in human urine using anion-exchange columns in HPLC-ICP-MS. PMID- 19542680 TI - Development of a work improvement checklist for occupational mental health focused on requests from workers. AB - OBJECTIVES: To develop tools offering definite orientation for managers and employees to support their work improvement through occupational mental health. This research was a part of the Mental Health Improvement & Reinforcement Study (MIR study), conducted from October 2004 to March 2006. METHODS: We developed a trial version named the Kaizen Check List (KCL) by referring to problem solving methods for quality management. Then we improved it for a formal version named MIR Research of Recognition (MIRROR). A feedback form named MIR Action Guidance (MIRAGe) was also developed. We analyzed data from 1,953 respondents at five manufacturing enterprises in Japan using MIRROR and the Brief Job Stress Questionnaire (BJSQ) to determine whether or not the workers requesting work improvement had more stress than other workers. RESULTS: The KCL had 47 items, which indicated desirable working conditions for mental health at work, and four answer categories. MIRROR has 45 selected items and improved answer categories. MIRAGe displays the results of MIRROR and step-by-step guidance for work improvement. Respondents with request had significantly higher scores in stressor and lower scores in buffer factors compared with respondents without request in many items of MIRROR. CONCLUSIONS: A combinational use of MIRROR and stress scales is useful for finding worksites with high risk factors for mental health and for directing focus on work improvement at these worksites according to workers' requests. PMID- 19542681 TI - The chondroprotective agent ITZ-1 inhibits interleukin-1beta-induced matrix metalloproteinase-13 production and suppresses nitric oxide-induced chondrocyte death. AB - In a screening program aimed at discovering anti-osteoarthritis (OA) drugs, we identified an imidazo[5,1-c][1,4]thiazine derivative, ITZ-1, that suppressed both interleukin-1beta (IL-1beta)-induced proteoglycan and collagen release from bovine nasal cartilage in vitro and suppressed intra-articular infusion of IL 1beta-induced cartilage proteoglycan degradation in rat knee joints. ITZ-1 did not inhibit enzyme activities of various matrix metalloproteinases (MMPs), which have pivotal roles in cartilage degradation, while it selectively inhibited IL 1beta-induced production of MMP-13 in human articular chondrocytes (HAC). IL 1beta-induced MMP production has been shown to be mediated by extracellular signal-regulated protein kinase (ERK), p38 kinase, and c-Jun N-terminal kinase (JNK) of the mitogen-activated protein kinase (MAPK) family signal transduction molecules. An ERK-MAPK pathway inhibitor (U0126), but not a p38 kinase inhibitor (SB203580) or a JNK inhibitor (SP600125), also selectively inhibited IL-1beta induced MMP-13 production in HAC. Furthermore, ITZ-1 selectively inhibited IL 1beta-induced ERK activation without affecting p38 kinase and JNK activation, which may account for its selective inhibition of MMP-13 production. Inhibition of nitric oxide (NO)-induced chondrocyte apoptosis has been another area of interest as a therapeutic strategy for OA, and ITZ-1 also suppressed NO-induced death in HAC. These results suggest that ITZ-1 is a promising lead compound for a disease modifying anti-OA drug program. PMID- 19542682 TI - Hypoxic ventilatory response in platelet-derived growth factor receptor-beta knockout mice. AB - The present study investigated whether the platelet-derived growth factor receptor (PDGFR)-beta-mediated mechanisms are involved in the hypoxic ventilatory response through modulating the N-methyl-D-aspartate (NMDA) function. The ventilatory changes during hypoxic challenge (10% O(2), 30 min) were measured plethysmographically in mice selectively lacking the PDGFR-beta in neurons (KO mice) and in control wild-type mice (WT mice) before and after blockade of NMDA receptors. In baseline breathing at rest, respiratory rate, tidal volume, and minute ventilation were similar between WT and KO mice. Hypoxia caused an increase of ventilation during the early period of exposure (an initial excitation), followed by a progressive decrease along with the exposure period (a late decline). The initial excitation occurred similarly in KO and WT mice, while the late decline was markedly attenuated in KO mice. Administration of an antagonist of NMDA receptors, dizocilpine (0.3 mg/kg, i.p.) decreased the initial excitation and hastened the late decline of hypoxic ventilatory response. Furthermore, the hypoxic ventilatory response in KO mice was indistinguishable from that in WT mice after blockade of NMDA receptors. The present study suggests that the PDGF-BB/PDGFR-beta signal axis contributes to the hypoxic ventilatory response by its inhibitory effect on the NMDA receptor-mediated function. PMID- 19542684 TI - Correlation of receptor occupancy of metabotropic glutamate receptor subtype 1 (mGluR1) in mouse brain with in vivo activity of allosteric mGluR1 antagonists. AB - The aim of this study was to clarify the relationship between receptor occupancy and in vivo pharmacological activity of mGluR1 antagonists. The tritiated mGluR1 selective allosteric antagonist [(3)H]FTIDC (4-[1-(2-fluoropyridin-3-yl)-5-methyl 1H-1,2,3-triazol-4-yl]-N-isopropyl-N-methyl-3,6-dihydropyridine-1(2H) carboxamide) was identified as a radioligand having high affinity for mGluR1 expressing CHO cells (K(D) = 2.1 nM) and mouse cerebellum (K(D) = 3.7 nM). [(3)H]FTIDC bound to mGluR1 was displaced by structurally unrelated allosteric antagonists, suggesting there is a mutual binding pocket shared with different allosteric antagonists. The binding specificity of [(3)H]FTIDC for mGluR1 in brain sections was demonstrated by the lack of significant binding to brain sections prepared from mGluR1-knockout mice. Ex vivo receptor occupancy with [(3)H]FTIDC revealed that the receptor occupancy level by FTIDC correlated well with FTIDC dosage and plasma concentration. Intracerebroventricular administration of (S)-3,5-dihydroxyphenylglycine is known to elicit face washing behavior that is mainly mediated by mGluR1. Inhibition of this behavioral change by FTIDC correlated with the receptor occupancy level of mGluR1 in the brain. A linear relationship between the receptor occupancy and in vivo activity was also demonstrated using structurally diverse mGluR1 antagonists. The receptor occupancy assays could help provide guidelines for selecting appropriate doses of allosteric mGluR1 antagonist for examining the function of mGluR1 in vivo. PMID- 19542683 TI - Polaprezinc (Zinc L-carnosine) is a potent inducer of anti-oxidative stress enzyme, heme oxygenase (HO)-1 - a new mechanism of gastric mucosal protection. AB - Heme oxygenase (HO)-1 is implicated in cytoprotection in various organs. We tested a possibility that polaprezinc (PZ), an anti-ulcer drug, could induce HO-1 in the gastric mucosa. Male 6-week-old Wistar rats were intragastrically administered PZ. Gastric expression of HO-1 was assessed by real time RT-PCR and western blotting, and localization of HO-1 was observed by in situ hybridization and immunohistochemistry. The levels of HO-1 mRNA were increased in a dose dependent manner. The levels of HO-1 mRNA were increased 4-fold by PZ at the dose of 200 mg/kg at 3 h as compared with control levels. The levels of immunoreactive HO-1 were increased 3-fold at 6 h. Signals for HO-1 mRNA and immunoreactivity were detected strongly in the surface gastric mucosal cells and moderately in the gastric macrophages. Treatment with an HO-1 inhibitor, stannous mesoporphyrin (SnMP) significantly worsened the HCl-induced acute gastric mucosal lesions and increased the apoptosis of mucosal cells. Mucosal lesions were decreased by pretreatment with PZ, while they were increased by co-administration with SnMP. These data indicate for the first time that PZ is an effective inducer of HO-1 in the stomach. PZ-induced HO-1 functions as a part of the mucosal protective effects of PZ. PMID- 19542685 TI - Change in mean height of Thai military recruits from 1972 through 2006. AB - BACKGROUND: Records in Western countries reveal that adult height has been increasing over the last 250 years. These height gains have been biologically associated with healthier childhoods, less illness, and longer life spans-a health-risk transition. To measure such progress in Thailand we studied height change over the last 3 decades. METHODS: We analyzed height records for 33 000 21 year-old male military recruits, sampling 1000 per year from 1972 through 2006. We compared the height trend in Thailand to those noted in Europe, and discuss the former in the context of improvements in living circumstances in Thailand. RESULTS: Over 35 years, mean height increased from 164.4 to 169.2 cm, an increment of nearly 5 cm. The height increase was negligible in the first decade (1972-1981), but substantially accelerated after that. In the period after 1990 the increase exceeded 3 cm. A similar overall height gain in Britain occurred over a much longer period (1750-1886). CONCLUSIONS: The increase in height among Thai men is biological evidence that a Thai health-risk transition-defined by both changing risks and outcomes-is well underway for height. Military recruits born during the 1960s through the 1980s had progressively healthier childhoods. Over this period child nutrition improved, infection and mortality rates declined, and preventive health services expanded. The combined effect of these factors is indicated by the increased adult height of Thai military recruits. PMID- 19542686 TI - End-stage renal disease in Taiwan: a case-control study. AB - BACKGROUND: Taiwan has the highest incidence of end-stage renal disease (ESRD) in the world. The epidemiologic features of ESRD, however, have not been investigated. In this case-control study, we evaluated the risk of ESRD associated with a number of putative risk factors. METHODS: We studied 200 patients among whom ESRD had been newly diagnosed between 1 January 2005 and 31 December 2005; 200 controls were selected from among relatives of patients treated in the general surgery unit. Using a structured questionnaire, we collected information related to socioeconomic factors, history of disease, regular blood or urine screening, lifestyle, environmental exposure, consumption of vitamin supplements, and regular drug use at 5 years before disease onset. RESULTS: Our primary multivariate risk models indicated that low socioeconomic status was a strong predictor of ESRD (education: odds ratio [OR], 2.78; 95% confidence interval [CI], 1.49-5.19; income: OR, 2.86, 95% CI, 1.48-5.52), even after adjusting for other risk factors. Other significant predictors for ESRD were a history of hypertension (OR, 3.63-3.90), history of diabetes (OR, 3.85 5.50), and regular intake of folk remedies or over-the-counter Chinese herbs (OR, 10.84-12.51). Regular intake of a multivitamin supplement 5 years before diagnosis was associated with a decreased risk of ESRD (OR, 0.12-0.14). CONCLUSIONS: Our findings indicate that low socioeconomic status, history of hypertension, diabetes, and regular use of folk remedies or over-the-counter Chinese herbs were significant risk factors for ESRD, while regular intake of a multivitamin supplement was associated with a decreased risk of ESRD. PMID- 19542688 TI - Subsite distribution of gastric cancer in an area of high prevalence--northwest Iran. AB - BACKGROUND: The aim of the present study was to determine subsites of gastric cancer in East Azerbaijan, Iran-a high incidence region for gastric cancer and Helicobacter pylori infection. METHODS: Data were collected from 2002 through 2007 from patients who sought treatment for gastrointestinal symptoms or signs at a university clinic and subsequently underwent upper gastrointestinal endoscopy. RESULTS: Cancer was diagnosed and histologically confirmed in 362 patients (352 adenocarcinomas). The mean age of the patients was 64.57 +/- 11.32 (range, 16-94 years) and the male-to-female ratio was 2.8:1. The gastric cardia was involved in 40.3% of patients with gastric adenocarcinoma, while the gastric fundus was involved in 3.7%, the gastric body in 49.1%, and the gastric antrum in 24.1% of patients. Complete evaluation for metastasis was possible in 144 patients; 61 were free of metastasis, and most of these patients underwent surgical therapy. Cardia involvement was not associated with the sex or age of patients. CONCLUSIONS: Noncardia gastric cancer is still more frequent in East Azerbaijan, which is likely due to the very high prevalence of infection with Helicobacter pylori. The low rate of cancer involving the fundus is a target for further research on the etiology of gastric cancer. PMID- 19542687 TI - Sociodemographic variation in the perception of barriers to exercise among Japanese adults. AB - BACKGROUND: The perception of barriers to exercise is an important correlate of exercise participation. However, only a limited number of studies-mostly from Western countries-have attempted to describe the perceptions of barriers to exercise in specific population groups. This study examined the associations between sociodemographic attributes and perceived barriers to exercise in Japanese adults. METHODS: A population-based cross sectional study of 865 participants (age: 20-69 years old, men: 46.5%) was conducted in 4 cities in Japan. Nine sociodemographic attributes (sex, age, location of residence, educational attainment, marital status, employment status, presence of dependents in the household, self-rated health, body mass index), along with exercise frequency and perception of barriers to exercise (discomfort, lack of motivation, lack of time, lack of social support, poor environment) were assessed by self administered questionnaire. RESULTS: The most strongly perceived barrier was lack of time. Five of 9 sociodemographic attributes were significantly related to certain types of perceived barriers. Participants who more strongly perceived barriers were younger, more highly educated, more likely to be employed, and had relatively poor self-rated health and a high BMI. The specific types of barriers that were strongly perceived varied with the sociodemographic attributes of the participants. CONCLUSIONS: The results show that the perception of barriers to exercise varies among specific population groups, which indicates the importance of targeting exercise promotion strategies to specific populations. PMID- 19542689 TI - Determination of (129)I and (127)I concentration in soil samples from the Chernobyl 30-km zone by AMS and ICP-MS. AB - A large amount of radioiodine isotopes (mainly (131)I, t(1/2) = 8 days) was released from the accident at Chernobyl Nuclear Power Plant (CNPP) in April-May 1986. An increase in childhood-thyroid cancer in the contaminated areas in Belarus, Russia and the Ukraine was demonstrated to be caused by radioiodine released at the time of the accident. However, there is a lack of quantitative data on the (131)I levels in the local environment (e.g. air, plant, soil). At this point, a long-lived iodine isotope, (129)I (t(1/2) = 15.7 million years), also released with a certain ratio to (131)I from CNPP, could be used for estimating the (131)I levels in the environment. In this paper we present analytical results of the (129)I concentrations and (129)I/(127)I atom ratios in soil samples collected from the CNPP exclusion zone (30-km zone), with the aim of assessing current contamination levels and distribution patterns. For the analysis of the iodine fraction in the investigated soil samples, the pyrohydrolysis method was utilized for separation of (127)I and (129)I nuclides, and subsequently their concentration was determined using inductively coupled plasma mass spectrometry (ICP-MS) and accelerator mass spectrometry (AMS), respectively. The concentration of (129)I and the (129)I/(127)I atom ratio in the surface soil samples in the 30 km-zone of CNPP ranged from 4.6 to 170 mBq/kg, and from 1.4 x 10(-6) to 13 x 10(-6), respectively. These values are significantly higher than those from global (129)I fallout, indicating that most of the measured (129)I was due to the deposition of the accident. Stable iodine concentrations in this area were found to be very low (below 1 ppm) for most of the samples, suggesting the environmental iodine levels in this area to be potentially low. The (129)I/(137)Cs activity ratio in surface and sub-surface soils was not so constant, i.e., in the range (7.3-20.2) x 10(-7). This might be due to the different behavior of deposition and/or migration of these nuclides in soil. These results suggest the obtained data of (129)I to be useful for the reconstruction of the (131)I deposition in the contaminated areas. PMID- 19542690 TI - Emerging connection between centrosome and DNA repair machinery. AB - Centrosomes function in proper cell division in animal cells. The centrosome consists of a pair of centrioles and the surrounding pericentriolar matrix (PCM). After cytokinesis, daughter cells each acquire one centrosome, which subsequently duplicates at the G1/S phase in a manner that is dependent upon CDK2/cyclin-E activity. Defects in the regulation of centrosome duplication lead to tumorigenesis through abnormal cell division and resulting inappropriate chromosome segregation. Therefore, maintenance of accurate centrosome number is important for cell fate. Excess number of centrosomes can be induced by several factors including ionizing radiation (IR). Recent studies have shown that several DNA repair proteins localize to the centrosome and are involved in the regulation of centrosome number possibly through cell cycle checkpoints or direct modification of centrosome proteins. Furthermore, it has been reported that the development of microcephaly is likely caused by defective expression of centrosome proteins, such as ASPM, which are also involved in the response to IR. The present review highlights centrosome duplication in association with genotoxic stresses and the regulatory mechanism mediated by DNA repair proteins.Translated and modified from Radiat. Biol. Res. Comm. Vol.43; 343-356 (2008.12, in Japanese). PMID- 19542691 TI - Targeted heavy-ion microbeam irradiation of the embryo but not yolk in the diapause-terminated egg of the silkworm, bombyx mori, induces the somatic mutation. AB - Using heavy-ion microbeam, we report target irradiation of selected compartments within the diapause-terminated egg and its mutational consequences in the silkworm, Bombyx mori. On one hand, carbon-ion exposure of embryo to 0.5-6 Gy increased the somatic mutation frequency, suggesting targeted radiation effects. On the other, such increases were not observed when yolk was targeted, suggesting a lack of nontargeted bystander effect. PMID- 19542692 TI - Bremsstrahlung and Photoneutron leakage from steel shielding board impinged by 12 24 MeV electrons beams. AB - Many medical linear accelerators generate not only high-energy photons, but also high-energy electrons, and they are no longer equipped with beam stoppers. Therefore, shields might be necessary against bremsstrahlung and photoneutron generated by high-energy electron beams. However, there are few physical studies, and no recommendations are made about shields nowadays. In this report, the leakage doses of bremsstrahlung and photoneutron were calculated by the use of Monte Carlo simulation. To verify the calculated results, the photoneutron leakage dose was measured with a rem counter. The results clearly show that the bremsstrahlung and photoneutron leakage dose generated by electron beams of 24 MeV or below is negligible. PMID- 19542693 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 1 Summary of findings in the 2008 UK Renal Registry Report. PMID- 19542694 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 7 Survival and causes of death of UK adult patients on renal replacement therapy in 2007: national and centre-specific analyses. AB - INTRODUCTION: These analyses examine survival from the start of renal replacement therapy (RRT), based on the total incident UK dialysis population reported to the Registry, including the 21% who started on PD and the 5% who received a pre emptive transplant. Survival of prevalent patients and changes in survival between 1997-2006 are reported. The article includes a discussion on the technical definition for the date of start of both PD and HD. METHODS: Survival was calculated for both incident and prevalent patients on RRT and compared between the UK countries after adjustment for age. Survival of incident patients (starting during 2006) was calculated with and without a 90 day RRT start cut off. Survival of incident patients is shown with and without censoring at transplantation. Both the Kaplan-Meier and Cox adjusted models were used to calculate survival. Causes of death were analysed for both groups. Relative risk of death was calculated compared with the general UK population. RESULTS: The 2006 unadjusted 1 year after 90 day survival for patients starting RRT was 86%. In incident 18-64 year olds the unadjusted 1 year survival had risen from 85.9% in 1997 to 91.5% in 2006 and for those aged > or = 65 it had risen from 63.8% to 72.9%. The age adjusted survival of prevalent dialysis patients rose from 85% in 2000 to 89% in 2007. Diabetic patient survival rose from 76.6% in 2000 to 84.0% in 2007. The relative risk of death on RRT compared with the general population was 30 at age 30 years compared with 3 at age 80 years. In the prevalent RRT dialysis population, cardiovascular disease accounted for 34% of deaths, infection 20% and treatment withdrawal 14%. CONCLUSIONS: Incident and prevalent patient survival on RRT in all the UK countries for all age ranges and also for patients with diabetes continued to improve. The relative risk of death on RRT compared with the general population has fallen since 2001. Death rates on dialysis in the UK remained lower than when compared with a similar aged population on dialysis in the USA. PMID- 19542695 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 3 ESRD incident rates in 2007 in the UK: national and centre-specific analyses. AB - INTRODUCTION: This chapter describes the characteristics of adult patients starting renal replacement therapy (RRT) in the UK in 2007 and the acceptance rate for RRT in Primary Care Trusts (PCT) or equivalent Health Authority (HA) areas in the UK. METHODS: The basic demographics are reported for all UK centres and clinical characteristics of patients starting RRT from all except 1 centre in the UK. Late presentation, defined as time between first being seen by a nephrologist and start of RRT being <90 days was also studied. Age and gender standardised ratios for acceptance rate in PCTs or equivalent HAs were calculated. RESULTS: In 2007, the acceptance rate in the UK was 109 per million population (pmp) compared to 111 pmp in 2006. Acceptance rates in England (107 pmp), Scotland (108 pmp) and Northern Ireland (105 pmp) have fallen slightly, whilst that in Wales (140 pmp) has risen. There were wide variations between PCTs/HAs with respect to the standardised ratios which were lower in more PCTs in the North West and South East of England and higher in London, the West Midlands and Wales. The median age of all incident patients was 64.1 years and for non Whites 57.1 years. There was an excess of males in all age groups starting RRT and nearly 80% of patients were reported to be White. Diabetic renal disease remained the single most common cause of renal failure (21.9%). By 90 days, 67.4% of patients were on haemodialysis, 21.3% on peritoneal dialysis, 5.2% had had a transplant and 6.1% had died or had stopped treatment. The incidence of late presentation in those centres supplying adequate data was 21%. CONCLUSIONS: The acceptance rate has fallen in England, Northern Ireland and Scotland but continues to rise in Wales with wide variations in acceptance rate between PCTs/HAs. PMID- 19542696 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 8 Adequacy of haemodialysis in UK renal centres in 2007: national and centre-specific analyses. AB - BACKGROUND: Outcome in patients treated with haemodialysis (HD) is influenced by the delivered dose of dialysis. The UK Renal Association (RA) publishes Clinical Practice Guidelines which include recommendations for dialysis dose. The urea reduction ratio (URR) is a widely used measure of dialysis dose. AIM: To determine the extent to which patients received the recommended dose of HD in the UK. METHODS: Seventy-one renal centres in the UK submit data electronically to the UK Renal Registry (UKRR). Two groups of patients were included in the analyses: the prevalent patient population on 31st December 2007 and the incident patient population for 2007. Centres returning data on <50% of their patient population were excluded from centre-specific comparisons. RESULTS: Data regarding URR were available from 61 renal centres in the UK. Forty six centres provided URR data on more than 90% of prevalent patients. 81% of prevalent HD patients met the UK Clinical Practice Guideline for URR (>65%) in 2007. There has been an increase from 56% in 1998 to 81% in 2007 in the proportion of patients in the UK who achieved a URR >65%. The HD dose (URR) delivered to patients who have just started dialysis treatment is lower than that of patients who have been treated for longer and increases further with time. CONCLUSIONS: The delivered dose of HD for patients with established renal failure has increased over 9 years. There was considerable variation from one centre to another, with 8 centres attaining the RA clinical practice guideline in >90% of patients and 7 centres attaining the standard in <60% of patients. PMID- 19542697 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 9 Haemoglobin, ferritin and erythropoietin amongst patients receiving dialysis in the UK in 2007: national and centre-specific analyses. AB - BACKGROUND: The UK Renal Association (RA) and National Institute for Health and Clinical Excellence (NICE) have published Clinical Practice Guidelines which include recommendations for management of anaemia in established renal failure. AIMS: To determine the extent to which the guidelines for anaemia management are met in the UK. METHODS: Quarterly data (haemoglobin (Hb) and factors that influence Hb) extracts from renal centres in England, Wales and Northern Ireland (EWNI), and annual data from the Scottish Renal Registry for incident and prevalent renal replacement therapy (RRT) cohorts for 2007 were analysed by the UK Renal Registry (UKRR). RESULTS: In the UK, in 2007 58% of patients commenced dialysis therapy with Hb > or = 10.0 g/dl (median Hb 10.3 g/dl). Of incident patients 81% and 87% had a Hb > or = 10.0 g/dl by 3 and 6 months of dialysis treatment respectively. The median Hb of haemodialysis (HD) patients was 11.6 g/dl with an interquartile range (IQR) of 10.6-12.6 g/dl. Of HD patients 86% had a Hb > or = 10.0 g/dl. The median Hb of peritoneal dialysis (PD) patients in the UK was 11.9 g/dl (IQR 11.0-12.8 g/dl). 91% of UK PD patients had a Hb > or = 10.0 g/dl. The median ferritin in HD patients in EWNI was 417 microg/L (IQR 270-598) and 95% of HD patients had a ferritin > or = 100 microg/L. The median ferritin in PD patients was 255 microg/L (IQR 143-411) with 85% of PD patients having a ferritin > or = 100 microg/L. In EWNI the mean ESA dose was higher for HD than PD patients (9,300 vs. 6,100 IU/week). CONCLUSIONS: This year for the first time there has been a small fall (from 85.9% in 2006 to 85.6%) in the percentage of HD patients with an Hb of > or = 10 g/dl. This contrasts with previous annual improvements in this figure and is related to implementation of the new Hb Standard which has a target range of 10.5-12.5 g/dl. PMID- 19542698 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 10 Biochemistry profile of patients receiving dialysis in the UK in 2007: national and centre specific analyses. AB - INTRODUCTION: The UK Renal Association Clinical Practice Guidelines include clinical performance measures for biochemical parameters in dialysis patients [1]. The UK Renal Registry (UKRR) annually audits dialysis centre performance against these measures as part of its role in promoting continuous quality improvement. METHODS: Cross sectional performance analyses were undertaken to compare dialysis centre achievement of clinical audit measures for prevalent haemodialysis (HD) and peritoneal dialysis (PD) cohorts in 2007. The biochemical variables studied were phosphate, adjusted calcium, parathyroid hormone, bicarbonate and total cholesterol. In addition longitudinal analyses were performed (2000-2007) to show changes in achievement of clinical performance measures over time. RESULTS: Serum phosphate was between 1.1-1.8 mmol/L in 53% of HD and 64% of PD patients. Since 2003 there has been annual improvement in phosphate control for both HD and PD patients, largely through a reduction in phosphate >1.8 mmol/L. PD patients this year also showed a reduction in the percentage with a low phosphate. Adjusted calcium was between 2.2-2.6 mmol/L in 73% of HD and 78% of PD patients. Parathyroid hormone was between 16-32 pmol/L in 25% of HD and 27% of PD patients. The audit measure for bicarbonate was achieved in 71% of HD and 50% of PD patients. There was inter-centre variation for all variables studied. CONCLUSIONS: The UKRR consistently demonstrates inter-centre variation in achievement of biochemical clinical audit measures. Understanding the causes of this variation is an important part of improving the care of dialysis patients in the UK. PMID- 19542699 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 11 Blood pressure profile of prevalent patients receiving dialysis in the UK in 2007: national and centre-specific analyses. AB - INTRODUCTION: Blood pressure (BP) control is assessed annually from patients on Renal Replacement Therapy at renal centres in England, Wales and Northern Ireland by the UK Renal Registry. METHODS: Patients alive and receiving RRT on 31st December 2007 with a BP reading in either the fourth or third quarter of 2007 were included. Summary statistics were calculated for each renal centre, nation and renal disease category. Linear regression analyses were performed for prevalent patients between 2000 and 2007. RESULTS: Significantly more haemodialysis patients achieved the BP standard (44.6% pre-HD and 48.8% post-HD) than peritoneal dialysis (32.8%) or renal transplant patients (26.7%). Median BP fell significantly between 2000 and 2007 for each treatment modality. There was significant variability in BP control between renal centres (p < 0.0001) for haemodialysis and transplant patients. Hypertension was significantly more common in haemodialysis patients with vascular disorders such as diabetes and renovascular disease (56.8%) than in glomerulonephritis (51.0%) or tubular disorders (45.1%). The effect was less prominent in peritoneal dialysis and not evident in transplant patients where few achieved the BP standard. CONCLUSION: A minority of patients on RRT achieved BP standards in 2007. There remained a significant variation in achievement of standards between renal centres. PMID- 19542700 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 12 Epidemiology of Methicillin Resistant Staphylococcus aureus bacteraemia amongst patients receiving Renal Replacement Therapy in England in 2007. AB - From April 2007, all centres providing Renal Replacement Therapy in England were asked to provide additional data on patients with Methicillin Resistant Staphylococcus aureus (MRSA) bacteraemia using a secure web-based system. Data were recorded on modality of treatment and the type of vascular access in use at diagnosis and in the previous 28 days. From April 2007 until March 2008, 188 discrete episodes of MRSA bacteraemia were reported in patients receiving dialysis for established renal failure. Over the same period 4,448 MRSA bacteraemias were reported in England, indicating that 4.2% of all cases occurred in dialysis patients. Of the 188 episodes, additional data from the renal centres were available in 92 cases (49%). All patients with completed records were on haemodialysis at the time of the bacteraemia. Of those, 65/92 (70.7%) were using venous catheters, the majority tunnelled lines (n = 55, 59.8%), and 2 other cases had used venous catheters in the previous 28 days. The relative risk of MRSA bacteraemia was about 100 fold higher for a dialysis patient in comparison to the general population and 8 fold higher for a patient using a catheter in comparison to a fistula. The mean rate for all patients was 0.92 +/- 0.85 episodes/100 prevalent dialysis patients/year but the rate varied between renal centres with a range of 0-3.28. Using just haemodialysis patients as the denominator, the mean was 1.14 +/- 0.95 episodes/100 patients/year with a range of 0-3.93. Compared to previous Registry reports, absolute numbers of reported MRSA bacteraemias has fallen by approximately 62% from 2004. Many centres have substantially reduced the numbers of cases. Dialysis patients are at increased risk of MRSA bacteraemia; this is closely associated with the use of venous catheters. The rate of MRSA bacteraemia is falling substantially within the prevalent dialysis population, but with variation in performance between centres. PMID- 19542701 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 13 Demography of the UK paediatric renal replacement therapy population. AB - AIMS: To describe the demographics of the paediatric RRT population in the UK and analyse changes in demographics with time. METHODS: Extraction and analysis of data from the UK paediatric Renal Registry. RESULTS: The UK paediatric established renal failure (ERF) population in April 2008 was 875 patients. The prevalence under the age of 16 years was 55 per million age related population (pmp) and the incidence 7.92 pmp. The incidence and prevalence for South Asian and Other ethnic groups were 3 times that of the White and Black populations. Renal dysplasia was the most common cause of ERF accounting for 33% of prevalent cases. Diseases with autosomal recessive inheritance were more common in patients from ethnic minority groups. The spectrum of diseases seen has changed over a generation. Overall 5 year survival for children with ERF was 91.8%. Five year survival of infants starting dialysis was just 62%. Transplanted patients accounted for 74% of the current population. The proportion with grafts from living donors has steadily risen to 34%. Children from ethnic minority groups were less likely to have an allograft and living donation was less frequent in this population. For those on dialysis, 57% were receiving peritoneal dialysis. This was the main treatment modality for patients under 4 years of age. CONCLUSIONS: The paediatric ERF population continued to expand slowly. Incidence and prevalence rates were stable and similar to other developed nations. The high incidence in patients from ethnic minority groups will lead to a greater proportion of the population being from these groups in time. To maintain the high proportion of engrafted patients it will be necessary to encourage living donation in the ethnic minority population. The spectrum of diseases seen has already changed over a generation with the treatment of young children with diseases such as congenital nephrosis. The incidence of cystinosis causing ERF was reduced, probably reflecting better early treatment. PMID- 19542702 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 14 UK Renal Registry and international comparisons. AB - BACKGROUND: The aim of this study is to report Renal Replacement Therapy (RRT) incidence and prevalence rates, the percentage of incident patients with diabetes mellitus as cause of renal disease, the RRT modality mix and the transplant rate in different countries. The number of national or regional registries collecting and reporting data pertaining to traditional cardiovascular (CV) risk factors in prevalent dialysis patients is also examined. METHODS: Data on numbers of incident and prevalent RRT patients in England, Wales, Scotland and Northern Ireland for the year 2007 were collected from the UK Renal Registry (UKRR) database and collated to meet the specifications on the US Renal Data System (USRDS) international data collection form. RESULTS: In 2007, the incidence and prevalence of RRT in the UK were 110 and 759 per million of the population (pmp) respectively. Incidence of RRT placed the UK 34th out of the 43 countries reporting to the USRDS in 2006. In the majority of reporting countries, 20-44% have diabetes as the primary cause of end stage renal disease. Only the Finnish, Malaysian and US Renal Registries were found to routinely report attainment of cardiovascular risk standards. CONCLUSIONS: A comparison among international renal registries about RRT epidemiology and reporting cardiovascular risk factors in prevalent RRT patients forms an important part of the quality improvement process and often allows for improving standards and performances between reporting countries. Despite the high CV morbidity associated with RRT, few renal registries routinely report data on CV risk management; where data are reported there is little agreement in what represents quality of care, making direct comparison difficult. PMID- 19542704 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 2 Introduction to the 2008 UK Renal Registry Report. PMID- 19542703 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 15 The UK Renal Registry, UKRR database, validation and methodology. AB - The UK Renal Registry receives encrypted data extracts quarterly from each centre providing Renal Replacement Therapy (RRT) in England, Wales and Northern Ireland. Summary data is received from the Scottish Renal Registry to allow national statistics to be compiled. Data from patients receiving haemodialysis in satellite units or at home are reported through the main renal centre. Data from patients with functioning kidney transplants are reported through the centre providing routine clinical follow-up. The data are extracted from a variety of IT systems with varying functionality and no common messaging system, necessitating extensive data validation and cleaning prior to analysis. Growing confidence in the analyses since the inception of the Registry in 1995 has allowed de anonymised centre-specific analyses of all outcomes, including survival, to be published, although incomplete data returns for primary renal diagnosis and comorbidity at start of RRT limit ability to adjust for case-mix. PMID- 19542705 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 4 ESRD prevalent rates in 2007 in the UK: national and centre-specific analyses. AB - INTRODUCTION: This chapter describes the demographics of UK RRT patients in 2007. METHODS: Complete data were electronically collected from 71 UK centres with the remaining 1 centre submitting summary data. A series of cross-sectional and longitudinal analyses were performed to describe the demographics of prevalent UK RRT patients in 2007 at a centre and a national level. RESULTS: There were 45,484 adult patients receiving RRT on 31/12/2007. The population prevalence for adults was 746 per million population per year (pmp) with an annual increase in prevalence of approximately 5% per annum. There was substantial variation in standardised prevalence ratios between Primary Care Trust (PCT)/Health Authority (HA) areas which were associated with geographical factors and differences in ethnicity with mean standardised prevalence ratios (SPR) significantly higher in PCTs/HAs with a high proportion of ethnic minorities. The median age of prevalent RRT patients was 57 years (HD 65 years, PD 60 years, transplant 50 years). Median RRT vintage was 5.3 years (HD 2.8 years, PD 2.1 years, transplant 10.4 years). For all ages, crude prevalence rates in males exceeded those in females, peaking in the 75-79 year age band for males at 2,506 pmp and in females in the 70-74 year age band at 1,314 pmp. The most common identifiable diagnosis was glomerulonephritis (15.3%) but in those over 65 it was diabetes (15.1%). The most common treatment modality was transplantation (46.6%), closely followed by centre based HD (42.1%) in either the primary centre (25.2%) or the satellite unit (16.9%). The HD population has continued to expand, and the PD population to contract. HD was increasingly prominent with increasing age at the expense of transplantation. CONCLUSIONS: There were national, area and dialysis centre level variation in the prevalent UK RRT population. This has implications for service planning and ensuring equity of care for RRT patients. PMID- 19542707 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 6 Comorbidities and current smoking status amongst patients starting renal replacement therapy in England, Wales and Northern Ireland: national and centre-specific analyses. AB - INTRODUCTION: The prevalence of 13 comorbid conditions and smoking status at the time of starting renal replacement therapy (RRT) in England, Wales and Northern Ireland are described. METHODS: Adult patients starting RRT between 2002 and 2007 in centres reporting to the UK Renal Registry (UKRR) and with data on comorbidity (n = 13,293) were included. The association of comorbidity with patient demographics, treatment modality, haemoglobin, renal function at start of RRT and subsequent listing for kidney transplantation were studied. Association between comorbidities and mortality at 90 days and one year after 90 days from start of RRT was explored using Cox regression. RESULTS: Completeness of data on comorbidity returned to the UKRR remained poor. Of patients with data, 52% had one or more comorbidities. Diabetes mellitus and ischaemic heart disease were the most common conditions seen in 28.9% and 22.5% of patients respectively. Comorbidities became more common with increasing age (up to the 65-74 age group), were more common amongst Whites and were associated with a lower likelihood of pre-emptive transplantation, a greater likelihood of starting on haemodialysis (rather than peritoneal dialysis) and a lower likelihood of being listed for kidney transplantation. In multivariable survival analysis, malignancy and ischaemic/neuropathic ulcers were the strongest predictors of poor survival at 1 year after 90 days from start of RRT. CONCLUSIONS: The majority of patients had at least one comorbid condition and comorbidity is an important predictor of early mortality on RRT. PMID- 19542706 TI - UK Renal Registry 11th Annual Report (December 2008): Chapter 5 Demographics and biochemistry profile of kidney transplant recipients in the UK in 2007: national and centre-specific analyses. AB - INTRODUCTION: Outcomes following renal transplantation are usually reported as graft or patient survival. However, graft function, haemoglobin and blood pressure are also important measures of quality of care. METHODS: Transplant activity and incident graft survival data were obtained from NHS Blood and Transplant (NHSBT), laboratory and clinical variables and prevalent survival data were obtained from the UK Renal Registry (UKRR). Data were analysed separately for prevalent and one year post-transplant patients. RESULTS: Increasing live and non-heartbeating donors were responsible for the increasing transplant activity. Transplant waiting list numbers continued to rise by 8%. Graft failure occurred in 3.2% of prevalent transplant patients. Death rates remained stable at 2.3/100 patient years. Malignancy accounted for 21% of these deaths. There was centre variation in outcomes such as eGFR and haemoglobin in prevalent and 1 year post transplant recipients. Analysis of prevalent transplants by chronic kidney disease stage showed 16% with eGFR <30 and 2.2% <15. Of those in stage 5T, 26% had Hb <10 g/dl, 27% phosphate > or = 1.8 mmol/L and 50% an iPTH > or = 32 pmol/L. These patients were less likely to achieve the UK standards in comparison to CKD5 dialysis patients. CONCLUSION: Wide variations in clinical and biochemical outcomes may be secondary to variations in the care administered to transplant recipients across the UK. PMID- 19542709 TI - [Cancer of unknown primary site-current status and future direction]. AB - Cancer of unknown primary site(CUP)is not a rare entity for 3-5% of all malignant neoplasms. CUPis diagnosed with metastatic lesion so they are all in the advanced stage. The diagnosis is made by biopsy. Patients with CUP need to have a physical examination, basic blood examination and appropriate diagnostic imaging. Those are valuable for final diagnosis of primary site determination. Systemic chemotherapy is applied in many cases, but the standard therapeutic strategy has not yet been determined. PMID- 19542708 TI - [Nomogram as predictive model in clinical practice]. AB - A nomogram which is developed based on logistic regression analysis with multiple factors provides accurate prediction in various situations. The ability of the nomograms to predict diagnosis, staging and prognosis in prostate cancer and other disease has been confirmed to be better than other predictive models such as risk stratification and artificial neural network. Making a nomogram requires a fixed number of patients and multiple steps such as validations and calibrations. And when nomograms are developed at other institutions, validations are essential for physicians before use at the actual clinical level. We review the clinical significance of nomograms and introduce the process of making a nomogram. PMID- 19542710 TI - [The pathological practice for metastatic carcinoma of unknown primary-the current status and future prospect]. AB - The pathological practice for metastatic carcinoma of unknown primary is mainly to decide the histological type and suggestive primary site, which are very important for the therapeutic approach. In addition, for the morphological features with ordinary hematoxylin-eosin(HE)staining, one of the most universal and important methods is immunohistochemical staining. In recent years, detailed investigations could be performed with the combination of various primary antibodies with high organ specificity, a different pattern or amount of expression in each organ, although some cases, especially those of poor to undifferentiated carcinomas, have remained unindentified. PMID- 19542711 TI - [Clinical diagnosis of primary unknown cancer-the present situation and problems]. AB - The first step of diagnosis of primary unknown cancer(PUC)the detailed history intake and physical examination including breast, genitourinary system and rectum. Laboratory test, chest X-p and systemic computed tomography are allowed to be performed for all patients with PUC. Other tests should be performed according to the results of clinical and pathological evaluation. Utility of the tumor marker is limited, and this test is not recommended as a routine usage. There is not enough evidence on the utility of FDG-PTT or FDG-PET/CT for patients with PUC. Diagnosis of PUC should be made within one month from a patient's first visit to a hospital. PMID- 19542712 TI - [Treatment of cancer of unknown primary, today and future]. AB - Cancer of unknown primary(CUP)is a malignant disease with metastases such as lymph nodes, lung or liver metastasis, and without an identified primary site. CUP constitutes 5% of all human malignant diseases. CUPs are divided into two sub groups, favorable and unfavorable, based on their clinical output. In the unfavorable group, platinumbased CUPs or those combined with taxane regimens have been examined in phase II trials and practically conducted with high frequency. However, no standard regimens have thus far been established. Chemotherapy based on the primary site is not applicable for the treatment of CUP. It is very difficult to apply any regimens without information on the primary sites. To resolve this problem, molecular diagnostic technologies using a gene expression profiling platform to identify the primary site of CUP are now applied. Primary site-dependent chemotherapy could be conducted in accordance with the result of the molecular diagnosis of the primary site. On the other hand, the idea that the unfavorable group is an independent entity of malignancy is supported. The unfavorable CUPs might have a common molecular mechanism. Identification of the underlying mechanism that is specific to the unfavorable CUP may be a clue to develop a novel treatment for CUP. PMID- 19542713 TI - [Molecular diagnostic methods designed for clinical approach to cancer of unknown origin]. AB - Several studies have shown that patterns of gene expression remain consistent with the tissue of origin in cancer samples. Gene expression profiling may therefore offer a promising new technology to build a site of origin classifier with the ultimate aim to determine the origin of cancer of unknown primary(CUP). A single cDNA microarray platform was used to profile 229 tumors of known origin(14 tumor types). This data set was subsequently used for training and validation of a support vector machine(SVM)classifier, demonstrating 89% accuracy to predict a site of origin(13 types). Applying this microarray SVM classifier to 13 cases of CUP, a high confidence prediction was made in 11 of 13 cases. These predictions were supported by comprehensive review of the patients' clinical histories. Thus, data generated using both microarray and quantitative PCR can be used to train and validate a cross-platform SVM model with high prediction accuracy. PMID- 19542714 TI - [Examination of s-1 therapy for adjuvant chemotherapy in patients with advanced gastric cancer]. AB - PURPOSE: In our department, S-1 has been administered for 1 year as postoperative adjuvant chemotherapy for advanced gastric cancer since 2000. It was started by a standard dosage of 4-week administration with 2 weeks rest since 2000(A group). However, since 2002, it was changed with the expectation of the reduction of side effects by 2-week administration with a one-week rest(B group). Treatment continuity, adverse events and efficacy in both A and B groups were examined. SUBJECTS: The subjects were 96 patients with fStage II, III A and III B who were treated with S-1 after curative operation between 2000 and 2006. RESULTS: The percentage of patients who complied with the dosing instructions completely during a 1-year period was 70.2% in the A group and 77.6% in the B group. The incidence of Grade 3 and 4 toxicity was 1 nausea, 2 appetite loss, 1 neutropenia, 1 liver dysfunction in the A group and 1 nausea, 3 neutropenia in the B group, against, 76.6% in the A group and 44.9% in the B group, respectively, in the case of discontinuation, Thus, the trend in significantly high incidence in the A group was recognized. The 3-year survival rates in cases with a case of over 3 years were 88.5% in the A group and 87.5% in the B group, i. e., no difference. CONCLUSION: Though the difference was not recognized in continuation rate, efficacy and adverse events in both A and B groups, there were significantly few withdrawal cases in the B group, and it seemed to be an effective medication method. PMID- 19542715 TI - [Transcatheter arterial chemoembolization for hepatocellular carcinoma using 3 French catheter system]. AB - PURPOSE: To describe the initial experience of transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC)using the 3 French(F)catheter system designed for visceral angiography. MATERIAL AND METHODS: Twenty patients with HCC underwent TACE via the right femoral artery using the combination of a 3 F sheath introducer, a 3.3 F catheter, and a 2-2.4 F microcatheter compatible with the 3.3 F catheter. Completion of homeostasis at the puncture site was assessed after five minutes of manual compression. We assessed whether the resumption of ambulating two hours after TACE was safe and feasible. RESULTS: In 18/20 patients(90%), selective TACE was achieved with the 3 F catheter system. In three of these patients, it was necessary to use a 4 F dilator before insertion of the 3 F sheath introducer. In two patients, the 3.3 F catheter could not enter the celiac artery, and needed to be changed to the 4 F catheter system. In one patient, it took eight minutes until homeostasis was completed. In sixteen patients, the scheduled resumption of ambulation was obtained. In contrast, two patients experienced rehemorrhage, and required further compression. CONCLUSION: TACE can be performed using the 3 F catheter system with early resumption of ambulating in most cases. PMID- 19542716 TI - [S-1 monotherapy in patients with pretreated advanced non-small cell lung cancer]. AB - BACKGROUND: The objective of this study was to evaluate the efficacy and toxicity of S-1 monotherapy in patients with pretreated advanced non-small cell lung cancer(NSCLC). METHODS: We prospectively analyzed patients previously treated with a platinum-containing regimen or monotherapy with a third-generation chemotherapeutic drug. S-1 was orally administered at approximately 80 mg/m / 2day for 28 days followed by a 2-week rest period. We evaluated the efficacy and toxicities. RESULTS: A total of 15 patients received S-1 monotherapy. Three partial responses were observed among them with an overall response rate of 20%. Toxicities of grade 3 or higher included anemia(13%), thrombocytopenia( 6%), fatigue(6%), anorexia(13%), diarrhea(13%), interstitialpneumonitis(6%)and infection(6%). The ratio of outpatient treatment period was 73.5%. The median time to progression, median survivaltime and 1-year survival rate were 4.2 months, 7.8 months and 27.8% respectively. CONCLUSIONS: S-1 monotherapy was suggested to be effective and tolerable in patients with pretreated advanced NSCLC. PMID- 19542717 TI - Prevention of venous pain and phlebitis caused by epirubicin hydrochloride. AB - Many patients complain of venous pain or develop phlebitis following treatment with epirubicin hydrochloride(EPI). To ensure effective and safe treatment with this drug, it is essential to deal with the adverse events associated with it appropriately. At our hospital, EPI was previously administered by drip infusion(diluted with 50mL of physiological saline)over 15 minutes after pretreatment(EPI main route). With this method of treatment, venous pain and phlebitis developed in 14 of 15 cases. In 3 of these 14 cases, the regimen was modified. Following this experience, EPI administration was switched to drip infusion from the fully-opened side tube used for pretreatment(EPI sub-route). Switching to this route resulted in a sharp decrease in the incidence of venous pain and phlebitis, to only 1 of 15 cases. Stimulation of vascular tunica intima by EPI has been considered a factor principally responsible for the venous pain and phlebitis seen after EPI therapy. To prevent these adverse reactions, it is necessary to modify the method of administration so that strong or long-term exposure of blood vessels to EPI can be reduced. The results of this study suggest that the EPI sub-route we devised is useful in achieving this goal. PMID- 19542718 TI - [A case of recurrent gastrointestinal stromal tumor of the stomach with complete response to imatinib mesilate]. AB - We report a patient who long had a complete response by chemotherapy with imatinib mesilate(IM)for locally recurrent gastrointestinal stromal tumor(GIST)of the stomach. On July 2000, a 58-year-old woman was pointed out to have anemia in the course of surveillance for malignant melanoma of skin. Endoscopic examination revealed continuous bleeding from a submucosal tumor with ulceration on the posterior wall of the stomach. After endoscopic homeostasis failed, emergency laparotomy was performed and a biopsy was also done. The diagnosis made was GIST from immunohistological findings of positive c-kit, positive CD34, negative HMB45, and negative S100. After diagnosis, total gastrectomy, distal pancreatectomy, and splenectomy were performed. On September 2003, a local recurrence was recognized, and then chemotherapy by 400 mg IM daily was started. After beginning with a dose of IM 400 mg daily, the reduction of the tumor was monitored. The IM dose then had to be reduced to 300 mg daily. Because of the adverse side effects of IM, systemic edema and body weight increased. After reduction of IM, the adverse reactions resolved promptly, and a complete response of the primary tumor has been maintained for 4 years 3 months. PMID- 19542719 TI - [Two advanced gastric cancer cases with peritoneal metastases successfully treated by s-1/paclitaxel combination therapy]. AB - Two unresectable advanced gastric cancer cases with peritoneal metastases were successfully treated by the combination therapy of S-1 and paclitaxel. S-1 (1.25m(2): 80 mg/day, 1.25m(2)-1.50m(2)<:120 mg/day) was administered orally for 14 consecutive days followed by 14 days rest and a 2-hour infusion of paclitaxel (50 mg/m(2)) was administered on day 1 and 15 of each course. Treatment was repeated every 4 weeks unless disease progression or severe adverse effects were observed. Case 1: 65-year-old male (performance status: PS 3) with type 1 gastric cancer with malignant ascites. Case 2: 66-year-old male (PS3) with peritoneal metastases whose primary gastric lesion was surgically resected. Partial response was obtained in the former and complete response in the latter. Combination therapy of S-1 and paclitaxel can be highly recommended for patients with inoperable gastric cancer with poor PS. PMID- 19542720 TI - [A case of marked response to CPT-11+CDDP neoadjuvant chemotherapy for advanced gastric cancer with paraaortic lymph node metastasis enabling curative resection and over 10-year survival]. AB - We report a case of advanced gastric carcinoma successfully treated with a combination of irinotecan and cisplatin as neoadjuvant chemotherapy. The patient, a 51-year-old woman, had a type 2 gastric carcinoma, which had metastasized to paraaortic lymph nodes. She was treated with irinotecan, 70 mg on day 1 and day 15, and cisplatin, 80 mg on day 1. The course was repeated every 4 weeks. Two courses of treatment resulted in a marked reduction of both the primary tumor and lymph nodes. Subsequently, the patient underwent curative surgery consisting of pancreatoduodenectomy and D3 lymph node dissection. No surgical complication was observed. On microscopic examination, only a few tumor cells were detected in the granulation tissues of the lymph nodes, but viable cancer cells still remained in the resected stomach. This patient has been free of disease for more than 10 years after operation. This case demonstrates the effectiveness and safety of irinotecan and cisplatin used in an neoadjuvant setting for treatment of advanced gastric carcinoma. PMID- 19542721 TI - [A case of multiple hepatic metastases of gastric cancer that showed complete regression by systemic chemotherapy using paclitaxel and UFT-E]. AB - We report a case of gastric cancer with simultaneous multiple liver metastasis that was successfully treated by paclitaxel and UFT-E. A 54-year-old man with gastric cancer was admitted to our hospital for further examination and treatment. A type III gastric cancer was located in the lower to middle part of the gastric body. Abdominal CT revealed multiple liver metastases and lymph node metastases. Then, we performed distal gastrectomy and cholecystectomy. Postoperative pathological diagnosis was stage IV(a type 3 tumor( 78x65 mm), pT3, por 2, INF g, ly3, v0, pN2(+)(26/ 28), H1(bilobular multiple metastases), CY0, P0). Postoperatively, he was treated with S-1 po at 100 mg/body/day as first-line chemotherapy. Thirteen days after S-1 initiation, he was readmitted due to grade 3 diarrhea, and S-1 was immediately stopped. After his general condition was improved, paclitaxel was administered biweekly at a dose of 80 mg/m2. He was discharged after twice administration, and the regimen was continued at an outpatient clinic. Four months after the operation, abdominal computed tomography(CT)showed a remarkable reduction of the multiple liver metastases, and the serum levels of tumor markers(CEA, CA19-9)were reduced. Five months after the operation, the serum levels of tumor markers elevated again. Then, additional administration of UFT-E po(300 mg/body daily) was started. Seven months after the operation, abdominal CT showed a complete regression of the multiple liver metastasis, and the serum levels of tumor markers were also reduced within the normal range. During chemotherapy at an outpatient clinic, critical adverse effects did not appear. Paclitaxel or paclitaxel combined with UFT-E might be an effective regimen as second- or third-line chemotherapy for the liver metastases of gastric cancer. PMID- 19542722 TI - [Local recurrence after surgical resection of pancreatic cancer effectively treated with combined chemoradiotherapy]. AB - We report a patient for whom systemic chemotherapy using gemcitabine was effective against local recurrence of pancreatic cancer. A 59-year-old man underwent pancreatoduodenectomy for pancreatic head cancer. The diagnosis was moderately-differentiated tubular adenocarcinoma(tubular type, pT2, pN0, fM0, fStage II ). Ten months after surgery, the patient had a CT examination which revealed a mass at the cut-end of the pancreas. The serum CA19-9 level was found to be elevated 790(U/mL). Chemotherapy with GEM(1,000 mg/m2)was administered intravenously on days 1, 8, and 15. Courses were repeated every 28 days. The patient received 8 courses of chemotherapy by GEM, and the regimen was changed to every two weeks because of the adverse event, leucopenia(grade 2)and thrombocytopenia( grade 2). Twenty-one months after chemotherapy, CT examination revealed regrowth at the same location at the cut-end of the pancreas, and so radiotherapy was performed at a total 63 Gy. The serum CA19-9 level dropped to within the normal range. The patient had been receiving systemic chemotherapy as an outpatient for 48 months without deterioration of quality of life. Unfortunately, the patient died of bacteriogenous meningitis 56 months after recurrence. Our experience suggests that this chemotherapy is simple and possible to continue safely on an ambulatory basis while maintaining quality of life. PMID- 19542723 TI - A case of advanced mucinous cystadenocarcinoma of the pancreas with peritoneal dissemination responding to gemcitabine. AB - Mucinous cystic neoplasm(MCN)of the pancreas is a rare disease. A 34-year-old female was referred to our hospital for a giant cystic tumor in the left epigastrium, suspected of being a pancreatic MCN. The surgical findings revealed that the tumor originated in the pancreatic tail with the presence of peritoneal dissemination. A distal pancreatectomy and a splenectomy were performed, and the resected specimen histologically revealed an invasive mucinous cystadenocarcinoma of the pancreas. The postoperative computed tomography(CT)scan showed metastatic tumors of the Douglas pouch and the left ovary. Gemcitabine(GEM)was thereafter systemically administered for palliative chemotherapy with a regimen of 1,000 mg/m / 2week for 3 weeks, followed by a week of rest. When assessed by a CT scan after 4 courses of chemotherapy, marked shrinkage of the tumors was identified, and we could not detect the tumors clearly. Moreover, the serum CA19-9 level fell from 341 U/mL to almost normal and there were no severe adverse events. Therefore, systemic chemotherapy with GEM is considered to possibly be an effective treatment against MCN. We describe herein the first case of advanced mucinous cystadenocarcinoma of the pancreas with peritoneal dissemination responding to GEM and a brief review of the literature. PMID- 19542724 TI - [A case of advanced ampullary carcinoma successfully resected after primary chemotherapy with s-1 and gemcitabine]. AB - The patient was a 53-year-old male who was admitted on an emergency basis with obstructive jaundice. He was diagnosed as having advanced ampullary carcinoma(T4 N0 H1, Stage IV b). To reduce the degree of obstructive jaundice, a self expandable metallic stent was placed in the area of the biliary obstruction. Immediately after relief from obstructive jaundice, combination chemotherapy of S 1 and gemcitabine was given. Subsequently, CA19-9, the tumormarker level was reduced, the metastatic liver tumor disappeared, and the size of the primary lesion was remarkably reduced. Therefore, a curative surgical resection was done. This is a very instructive case for developing effective chemotherapy options to treat biliary tract cancers involving ampullary carcinoma. PMID- 19542725 TI - [Two cases of KRAS wild-type unresectable or recurrent colorectal cancer effectively treated by cetuximab after progression of prior chemotherapy]. AB - Cetuximab, an anti-EGFR human/mouse chimeric antibody, has just been approved in Japan for patients with EGFR positive unresectable or recurrent colorectal cancer, as monotherapy or in combination with irinotecan. We reported two cases of unresectable or recurrent colorectal cancer effectively treated by cetuximab after the progression of the prior chemotherapy. Case 1: A51-year-old male suffered from sigmoid colon cancer with synchronous liver metastases. He received cetuximab plus irinotecan combination therapy in the third-line setting. Amonth after the initiation of the chemotherapy, abdominal CT showed tumor shrinkage of liver metastases. Case 2: A57-year-old female suffered from sigmoid colon cancer with metachronous liver, ovarian metastases, ascites and pleural effusion. Her performance status(PS)according to ECOG performance scale was 1, and she complained of dyspnea on exertion. She received cetuximab monotherapy in the fourth-line setting. Five weeks after initiation of chemotherapy, her chest, abdominal and pelvic CT showed tumor shrinkage of the liver metastases and the reduction of both ascites and pleural effusion, together with resolution of her dyspnea on exertion. Before cetuximab administration, we investigated KRAS status on cancer tissue previously resected in the above 2 cases, which showed KRAS wild type. Cetuximab could be effective for KRAS wild-type colorectal cancer, as well as the previous reports from Western countries. PMID- 19542726 TI - [A case of retroperitoneal tumor successfully resected thanks to effective chemotherapy]. AB - We reported a case of retroperitoneal immature teratoma with embryonal carcinoma. First, pathological diagnosis of sarcoma was made by fine needle aspiration, then secondary pathological diagnosis of suspected malignant schwannoma was made by a partial resection. The final pathological diagnosis by operation was immature teratoma with embryonal carcinoma. The preoperative diagnosis was difficult in this case, and the tumor had grown too large after partial resection to surgically resect. The effective chemotherapy reduced this tumor enough to allow successful resection. Although the prognosis of immature retroperitoneal teratoma is poor, this case has survived 10 years after operation without metastasis and recurrence. PMID- 19542727 TI - [Efficacy of treatment with high-dose toremifene in patients with metastatic breast cancer resistant to aromatase inhibitor treatment]. AB - The efficacy of aromatase inhibitors(AI)has been established for adjuvant and metastatic breast cancer. However, decision making regarding treatment becomes difficult after AI treatment. Recently, high-dose toremifene(HD-TOR, TOR 120 mg daily)showed efficacy in these patients. We attempted to study retrospectively the efficacy and safety of HD-TOR treatment. Seven patients received HD-TOR. The overall response rate was 29%(PR 2)and clinical benefit (CR, PR, long SD)was 57%(PR 2, long SD 7). HD-TOR may be an optional treatment for MBC after AI treatment. PMID- 19542728 TI - [A case of spindle cell carcinoma of the breast including metaplastic lesion with poor prognosis]. AB - A 57-year-old woman complained of a huge and rapid growing mass with bleeding in the left breast. Breast imaging (CT and MRI)showed a large, irregular and unevenly enhanced tumor with lymph node swelling in the left axilla. Mastectomy and axillary lymph node dissection were performed for control of bleeding from the tumor in the left breast. Pathological diagnosis was spindle cell carcinoma of the breast with transition from papillotubular carcinoma. Although the patient was treated with adjuvant chemotherapy and trastuzumab according to the treatment guideline for conventional breast cancer, she had an early relapse with mediastinal metastasis and died 9 months after operation. The tumor showed metaplastic change from epithelial tumor to spindle cell carcinoma. Because the epithelial part expressed weakly positive estrogen receptor(ER), progesterone receptor(PgR)-negative and HER2-positive, we used trastuzumab for adjuvant therapy. However, part of the spindle cell tumor mainly showed triple-negative, and ER, PgR and HER2 expression were negative, which might explain her poor prognosis for resistance to trastuzumab. PMID- 19542729 TI - [A case of anaplastic carcinoma of thyroid administered peroral fluorinated pyrimidine for long-term survival during three years]. AB - We report a case of anaplastic carcinoma of the thyroid administered peroral fluorinated pyrimidine, providing longterm survival during three years. Three years ago, a 70-year-old woman diagnosed with anaplastic carcinoma of the thyroid was admitted for chemoradiation therapy after tumor resection and tracheostomy. The tumor was confirmed to remain on the margin of resected specimens by postoperative histopathology. Combination chemotherapy, including docetaxel, 5 fluorouracil, cisplatin, and radiotherapy, was performed for four weeks. After completion of radiotherapy, CT scan showed residual tumor and lytic clavicular bone findings. But the patient did not desire the same heavy chemotherapy, wishing rather to undergo chemotherapy as an outpatient, if possible. She received chemotherapy with S-1 which was peroral fluorinated pyrimidine for 9 months. In this period, PET-CT showed that accumulation of FDG and lytic clavicular bone findings were continuously confirmed. She was changed to tegafur uracil(UFT)from S-1, because of myelosuppression. She is alive at this writing in good physical condition. CT shows no tumor growth and no remote metastasis. PMID- 19542730 TI - [Safe and successful chemoradiotherapy for a patient with cardiac pacemaker and triple cancers]. AB - The patient was a 70-year-old male with superficial hypopharyngeal cancer and advanced cancers of the esophagus and stomach. In his past history, a cardiac pacemaker was implanted for sick sinus syndrome. Further examination showed esophageal cancer had metastasized to the cervical lymph nodes and invaded the trachea. There was no surgical indication. In terms of chemoradiotherapy, it was thought to be possible because the patient was not pacemaker- dependent and radiotherapy could be planned in such a way as to keep the dose to the pacemaker as low as possible. Electrocardiogram was monitored during the treatment. In addition to the usual observation, the patient's cardiac symptoms and pacemaker status were assessed before and soon after the completion of radiotherapy. S-1 was selected as a concomitant chemotherapy. In liaising with the cardiologist and radiologist, chemoradiotherapy was achieved without pacemaker malfunction, and shrinking of tumors was also detected. PMID- 19542731 TI - [Sorafenib(Nexavar)]. AB - Sorafenib(Nexavar)is a multikinase inhibitor, with disruptive activity at intracellular C-RAF, B-RAF and mutant BRAF receptors, and extracellular C-KIT, FLT-3, VEGFR-2, VEGFR-3 and PDGFRb receptors. In the phase III study, as compared with placebo, treatment with sorafenib significantly prolonged progression free survival(PFS)in patients with advanced renal cell carcinoma in whom previous therapy has failed. Diarrhea, rash, fatigue, hand-foot skin reactions, and hypertension were the most common adverse events associated with sorafenib. As sorafenib was associated with similar rates of clinically manageable side effects in elderly patients as compared to younger patients, response rates to sorafenib in elderly patients were comparable to those of younger patients. Sorafenib was approved multinationally for the treatment of advanced and/or metastatic renal cell carcinoma. Sorafenib and sunitinib are reference standards of care for the treatment of advanced renal cell carcinoma and are recommended by current clinical guidelines. For the future, research of biomarker, adverse drug reaction, and combined regimens are needed to maximize the effects of molecular targeted drugs. PMID- 19542732 TI - [Future prospects of neoadjuvant endocrine therapy in postmenopausal breast cancer]. AB - Third-generation aromatase inhibitors(AIs)have recently overtaken tamoxifen(TAM)to play a key role in the adjuvant treatment of post-menopausal hormone receptor-positive breast cancer patients. The accumulating evidence of neoadjuvant chemotherapy is now contributing to growing interest in neoadjuvant endocrine therapy using AIs. Recent research has revealed that administration of AIs 3-4 months prior to surgery is more effective than TAM in terms of the clinical response rate and the breast-conserving surgical procedure rate. In addition, the Preoperative Endocrine Prognostic Index(PEPI), which uses four factors, namely MIB1/Ki67 levels, pathological tumor size, and lymph node and ER status following neoadjuvant endocrine therapy, has been demonstrated very effective for predicting the prognosis of patients treated with AIs. Investigation of the different efficacies of various AIs, the optimal dosing period, and suitable combination with adjuvant chemotherapy will enable breast cancer drug therapy to be individualized to suit each patient's condition. PMID- 19542735 TI - Efficacy of the new long-acting formulation of lanreotide (lanreotide Autogel) in somatostatin analogue-naive patients with acromegaly. AB - OBJECTIVE: To evaluate efficacy and safety of lanreotide autogel (ATG) 120 mg injections every 4-8 weeks in somatostatin analogue-naive patients with acromegaly. DESIGN: Open, non-comparative, phase III, multicenter clinical study. METHODS: Fifty-one patients (28 women, aged 19-78 yr): 39 newly diagnosed (de novo) and 12 who had previously undergone unsuccessful surgery (post-op, 11 macro and 1 micro) were studied. ATG 120 mg was initially given every 8 weeks for 24 weeks and subsequently changed according to GH levels: if 5 microg/l every 4 weeks (group C, 19 patients). Treatment duration was 48-52 weeks. The primary objective was to control GH and IGF-I levels (GH0.05). In Groups D and I, the UA levels showed a negative correlation with GIR. No significant difference was observed in the levels of 25-hydroxyvitamin D [25-(OH)D]. The levels of 1,25 (OH)2D in Groups D and T1 were markedly lower than that of Groups N or T2 (p<0.01). The 1,25-(OH)2D level in Group I was lower than that of Group N (p<0.05), but higher than that of Group D (p<0.01). The 1,25-(OH)2D level in Group T2 was nearly equivalent to that of Group N. In Groups D and I, the levels of 1,25-(OH)2D were negatively correlated with UA, and positively correlated with GIR. The BMD levels in lumbar vertebrae or femoral bone in Groups D and T1 were similar, but both were lower than that of Groups T2 (p<0.05) and N (p<0.01). The BMD levels were lower in Groups I and T2 compared with that of Group N (p<0.05), but higher than that of Groups D and T1 (p<0.05). The BMD levels in lumbar vertebrae or femoral bone in Groups I and D were positively correlated with GIR. The BMD level in lumbar vertebrae or femoral bone in Group D showed negative correlation with UA. CONCLUSION: In elderly rats with T2DM or IR, renal injury may cause decreased activity of renal 1-alpha hydroxylase, which may result in bone loss and disturbance in VD metabolism, mainly manifesting as a significant reduction in the 1,25-(OH)2D level. PMID- 19542736 TI - Increased clearance of cortisol by 5beta-reductase in a subgroup of women with adrenal hyperandrogenism in polycystic ovary syndrome. AB - OBJECTIVE: Increased peripheral metabolism of cortisol may explain compensatory ACTH-dependent adrenal steroidogenesis and hence hyperandrogenism in polycystic ovary syndrome (PCOS). Previous studies have described an increased 5alpha reduction of cortisol or impaired regeneration of cortisol by 11beta-HSD1 in PCOS. However, these observations may be confounded by obesity. Moreover, the relationship between alterations in cortisol metabolism and the extent of adrenal androgen hyper-secretion in response to ACTH has not been established. This study aimed to examine the association between cortisol metabolism and ACTH-dependent adrenal hyperandrogenism in PCOS, independently of obesity. DESIGN: We compared 90 PCOS women (age 18-45 yr) stratified by adrenal androgen responses to ACTH1-24 and 45 controls matched for age and body weight. METHODS: PCOS women were stratified as normal responders (NR), intermediate responders (IR), and high responders (HR) to 250 microg ACTH1-24: NR (no.=27) had androstenedione and DHEA responses within 2 SD of the mean in controls; IR (no.=43) had DHEA responses >2 SD above controls; HR (no.=20) had both androstenedione and DHEA responses >2 SD above controls. RESULTS: All groups were similar for age, body weight, and body fat distribution. Basal testosterone, androstenedione, and 5alpha dihydrotestosterone plasma levels were similarly elevated among the 3 groups of PCOS compared with controls, whereas basal DHEA-S was higher in HR (2.8+/-1.2 microg/ml) and IR (2.4+/-1.1 microg/ml) than in NR (1.8+/-0.8 microg/ml) and controls (1.7+/-0.6 microg/ml). The HR group had the lowest basal plasma cortisol levels (101+/-36 ng/ml vs IR 135+/-42 ng/ml, NR 144+/-48 ng/ml, and controls 165+/-48 ng/ml; all p<0.01), but the greatest cortisol response to ACTH1-24 (Delta(60-0)cortisol 173+/-60 ng/ml vs IR 136+/-51 ng/ml, NR 114+/-50 ng/ml, and controls 127+/-50 ng/ml; all p<0.01), and the highest urinary excretion of total and 5beta-reduced cortisol metabolites (eg 5beta-tetrahydrocortisol/ cortisol ratio 25.2+/-15.3 vs IR 18.8+/-10.7, NR 19.7+/-11.4, and controls 17.2+/-13.7; all p<0.05). There were no differences in urinary excretion of 5alpha-reduced cortisol metabolites or in 5alpha-dihydrotestosterone/testosterone ratio between groups. CONCLUSIONS: Adrenal androgen excess in PCOS is associated with increased inactivation of cortisol by 5beta-reductase that may lower cortisol blood levels and stimulate ACTH-dependent steroidogenesis. PMID- 19542737 TI - The relationship of the apolipoprotein E gene polymorphism Turkish Type 2 diabetic patients with and without nephropathy. AB - OBJECTIVE: Apolipoprotein E (ApoE) genetic variation which is a major constituent of plasma lipoproteins causes diabetic nephropathy progress. Chronic kidney disease is associated with increased E2 allele and the decreased E4 allele risk. The aim of this study was to investigate the association between ApoE gene polymorphism in the development of diabetic nephropathy in Type 2 diabetes Turkish patients. RESEARCH DESIGN AND METHODS: The objective of the study is to investigate the influence of ApoE gene polymorphism in the development of diabetic nephropathy in Turkish Type 2 diabetes. The ApoE genotypes were determined retrospectively in 46 patients with nephropathy and 56 without nephropathy and a control group of 35 healthy individuals. Genomic DNA was extracted from peripheral leukocytes of the subjects using the High Pure PCR Template Preparation Kit. For the detection of the presence of the three ApoE E alleles epsilon2, epsilon3, and epsilon4 (codon 112 and 158) were analyzed by the commercial LightCycler ApoE Mutation Detection Kit. RESULTS: No differences in ApoE genotype or the allelic frequencies of epsilon2, epsilon3 or epsilon4 were found between the Type 2 diabetic patient group (with and without nephropathy) and a control group. CONCLUSIONS: We conclude that the ApoE gene polymorphism is not associated with the development of diabetic nephropathy in Turkish Type 2 diabetic patients. Lack of association between ApoE gene polymorphism and Type 2 diabetic nephropathy might be due to ethnic differences. PMID- 19542738 TI - Prevalence of Y microdeletions in azoospermic and severe oligozoospermic men in Southern Italy: application of a rapid capillary electrophoresis method. AB - Male infertility is correlated with several genetic and non-genetic conditions. Microdeletions of Y chromosome are one of the most frequent genetic defects associated with male infertility. Evaluating this in infertile patients is important to assess an etiological diagnosis and possible prognosis of infertility, as well as to address clinical decision during treatment of infertility by intracytoplasmatic sperm injection, where the probability of success depends on the type and the number of deleted regions (azoospermia factor regions). To improve genetic counseling, it is useful to characterize Yq regions by a rapid and accurate method. In the current study, we evaluated the diagnostic efficiency and the time required of an in-house automated capillary electrophoresis method for Y microdeletions screening and applied it to estimate the prevalence of Y microdeletions in 100 infertile males affected by azoospermia or severe oligozoospermia (sperm count <5x10(6)/ml) and in 100 fertile male controls. In south Italian infertile men, the overall frequency of Y microdeletions was 9% (12.7% in azoospermic and 4.5% in severe oligozoospermic men). In conclusion, we think that the abovementioned procedure is suitable for the routine characterization of Y microdeletions. PMID- 19542739 TI - Effectiveness of retinoic acid treatment for redifferentiation of thyroid cancer in relation to recovery of radioiodine uptake. AB - BACKGROUND: Retinoic acid (RA) treatment has been used for redifferentiation of metastatic thyroid neoplasia that have lost radioiodine (131I) uptake with heterogeneous results. AIM: Retrospective analysis of the recovery rate of 131I uptake after RA treatment in patients from 11 Spanish hospitals. METHODS: Twenty seven patients (14 men, 13 women) with papillary [21], follicular [4], and oncocytic [2] thyroid cancer initially treated with total thyroidectomy plus 131I, and with 131I negative metastatic disease, were given 13-cis RA (0.66-1.5 mg/kg for 5-12 weeks) followed by a therapeutic 131I dose (3700-7400 MBq); 3 months later thyroglobulin levels and computed tomography imaging were performed. RESULTS: In 9 out 27 cases (33%) (8 papillary, 1 follicular) optimal positive 131I scan was observed after RA treatment; in the remaining 18, 10 had a suboptimal uptake (7 papillary, 2 follicular, 1 oncocytic) and in the rest there was no 131I uptake recovery (6 papillary, 1 follicular, 1 oncocytic). In 17 positive responses to RA (either optimal or suboptimal) in which image follow-up was available, decrease or stabilization of metastatic growth was observed in 7, while tumor mass increased at short term in the remaining 10. No major side effects were detected. CONCLUSION: Quite a high rate of 131I uptake recovery after RA treatment may be obtained in advanced differentiated thyroid cancer, but the potential modification of the natural course of the disease is uncertain. A better biological characterization of these tumors allowing the identification of potential responders to RA may improve the outcome of RA coadjuvant therapy. PMID- 19542740 TI - Degree of catecholamine hypersecretion is the most important determinant of intra operative hemodynamic outcomes in pheochromocytoma. AB - Pheochromocytoma resection is often complicated by intra-operative hypertension and post-resection hypotension. Factors associated with these hemodynamic alterations are not well defined. The aim of this study was to analyse the clinical-laboratory features associated with hemodynamic parameters during pheochromocytoma resection. Twenty-seven patients submitted to tumor resection - either open (no.=18) or video laparoscopic - between 1978-2007 were included. Nineteen received pre-operative alpha-blockers. Intra-operative hemodynamic data analysed were: maximum and minimum mean arterial blood pressure (MABP), no. of severe hypertensive (systolic BP >200 mmHg) and hypotensive episodes (MABP <60 mmHg), maximum and minimum heart rate (HR), no. of episodes of tachycardia and bradycardia, need to receive iv intra-operative treatment for hypertension and hypotension and the volume of fluids administered during surgery. Patients were 39.4+/-14.4-yr-old, 66% women. Intra-operative hemodynamic parameters were not different in patients submitted to open or video laparoscopic resection. Maximum intraoperative HR and the percentage of patients with HR>100 beats/min were higher in patients without pre-operative alpha- blocker treatment (no.=8). Pre operative urinary vanylmandelic acid was positively associated with intra operative maximum MABP (r=0.535, p=0.047) and with maximum transoperative systolic BP (r=0.805, p=0.016). Pre-operative urinary catecholamine (Pearson correlation r=0.575, p=0.03) and vanylmandelic acid (Pearson correlation r=0.605, p=0.04) levels were associated with maximum intra- operative MABP, adjusted for the presence of pheochromocytoma symptoms, surgical approach and pre-operative alpha-blockers. In conclusion, the degree of pre-operative catecholamine secretion was the most important aspect of transoperative BP control. PMID- 19542741 TI - Mutations in TAZ/WWTR1, a co-activator of NKX2.1 and PAX8 are not a frequent cause of thyroid dysgenesis. AB - AIM: In 80-85% of cases, congenital hypothyroidism is associated with thyroid dysgenesis (TD), but only in a small percentage of cases mutations in thyroid transcription factors (NKX2.1, PAX8, FOXE1, and NKX2.5) have been associated with the disease. Several studies demonstrated that the activity of the transcription factors can be modulated by the interaction with other proteins, such as coactivators and co-repressors, and TAZ (transcriptional co-activator with PDZ binding motif or WWTR1) is a co-activator interacting with both NKX2.1 and PAX8. In the present study we investigate the role of TAZ in the pathogenesis of TD. MATERIAL AND METHODS: By Single Stranded Conformational Polymorphism, we screened the entire TAZ coding sequence for mutations in 96 patients with TD and in 96 normal controls. RESULTS: No mutations were found in patients and controls, but we found several polymorphisms in both groups. No significant differences could be demonstrated in the prevalence of the mutations between patients and controls. CONCLUSIONS: Our data indicate that TAZ mutations are not a cause of TD in the series of patients studied. PMID- 19542742 TI - Effects of recombinant human relaxin upon proliferation of cardiac fibroblast and synthesis of collagen under high glucose condition. AB - Cardiac fibrosis is a key component of diabetes and involves the proliferation and differentiation of matrix-producing fibroblasts. We determined the influence of high glucose (HG) conditions on cardiac fibroblasts (CF) functions and the effects of recombinant human (rh) relaxin (RLX) in these responses. We cultured neonatal rat CF in either normal glucose (NG) or HG media. The mRNA of procollagen types I and III, and RLX-1 were assessed by real time PCR and procollagen type I C-terminal peptide (PICP) and procollagen type III amino terminal peptide (PIIINP), matrix metalloproteinases 2 (MMP2), MMP9 were assessed by enzyme linked immunosorbent assay. The results are as follows: a) CF proliferation was significantly increased by HG; rhRLX significantly inhibited HG fibroblast proliferation, while it had no marked effect on CF proliferation in NG. b) CF treated with HG significantly increased the production of PICP and PIIINP. rhRLX had no marked effect on production of PICP and PIIINP in NG. rhRLX blocked the HG-induced increases in collagen synthesis. c) The production of MMP2 and MMP9 is significantly increased by HG. rhRLX decreased overproduction of MMP2 and MMP9 in the presence of HG. d) The RLX- 1 mRNA expression of HG group was higher than in the NG group. We concluded that rhRLX could inhibit both the proliferation of CF and the synthesis of collagen under the HG condition. HG concentration could stimulate the expression of endogenous RLX. PMID- 19542743 TI - Assessment of left ventricular functions by tissue Doppler echocardiography in patients with Cushing's disease. AB - OBJECTIVE: To verify whether tissue Doppler imaging (TDI) could contribute to a better understanding of the natural history of cardiomyopathy in active Cushing's disease (CD), through its enhanced sensitivity to diastolic dysfunction, and identifying preliminary regional signs of systolic dysfunction before the appearance of clinical symptoms of cardiac pathologies. METHODS: Eleven women with newly diagnosed CD and 32 control cases, purposely matched for gender, age, body mass index and co-incidental diseases were enrolled in this study. Echocardiographic examinations were assessed by conventional echocardiography and tissue Doppler imaging. The peak systolic velocity (S'm), early diastolic myocardial peak velocity (E'm), late diastolic myocardial peak velocity (A'm), isovolumic acceleration (IVA), myocardial pre-contraction time (PCT'm), myocardial contraction time (CT'm) and myocardial relaxation time (RT'm) were measured at septal and lateral mitral anulus. RESULTS: In TDI, E'm and, E'm/A'm ratio were significantly lower, and PCT'm/CT'm ratio was higher, S'm, A'm, peak early diastole/E'm ratio, PCT'm, and isovolumetric myocardial relaxation time values were similar at lateral and septal anulus in patients with CD than controls (p>0.05). Lateral and septal anulus IVA were significantly lower in patients with CD than the control group (p<0.05). Correlation analysis showed that IVA time at lateral anulus correlated positively with S'm at lateral anulus (r=0.58; p=0.002) and IVA time at septal anulus correlated positively with S'm at septal anulus (r=0.51; p=0.008). CONCLUSION: Our study confirms that patients with CD have impaired diastolic function. More importantly, we also demonstrated an impairment of myocardial systolic function in patients with CD by TDI. We recommend using TDI in addition to conventional echocardiography parameters for the cardiovascular risk assessment of patients with Cushing' syndrome. PMID- 19542744 TI - Thyroid hormone predisposes rabbits to atrial arrhythmias by shortening monophasic action period and effective refractory period: results from an in vivo study. AB - BACKGROUND: Atrial arrhythmias are common complications of hyperthyroidism, but the underlying mechanisms remain to be further clarified. Thus, in this study, we try to investigate the effects of thyroid hormone on atrial electrophysiology by using a hyperthyroidism model in vivo. MATERIALS AND METHODS: Twenty-four New Zealand white rabbits were randomized into Thyroxine group (no.=12) and Control group (no.=12). In Thyroxine group, Levo-thyroxine (L-T(4)) solution (1 mg/kg x d(-1)) was injected daily into the peritoneum for 2 weeks. In Control group, the same amount of saline was injected. On day 15, 8 rabbits in each group were chosen randomly to receive electrophysiological experiment in vivo, in which electrophysiological parameters and atrial arrhythmias induced by electrical stimulation were recorded and serum thyroid hormone levels were examined. The others were killed so as to exam the L-type calcium current of atrium. RESULTS: Atrial monophasic action potential at 90 repolarization (AMAP(90)) and effective refractory period (AERP) were significantly shorter in Thyroxine group than in Control group (AMAP(90): 103.21+/-1.94 vs 122.14+/-6.13, p<0.01; AERP: 82.69+/ 0.99 vs 102.46+/-2.32, p<0.01). There are significant differences in the incidence of atrial arrhythmias between the two groups. The mean peak of L-type calcium current (I(Ca,L)) densities (pA/pF) at -10mV was significantly higher in Thyroxine group than in Control group (-8.59+/-0.68 vs -6.54+/-0.49, no.=8, p<0.001). CONCLUSIONS: In our hyperthyroidism model, thyroid hormone predisposed rabbits to atrial arrhythmias by shortening AMAP and AERP, which might be associated with increased I(Ca,L) current densities in atrium. PMID- 19542745 TI - Association of 1704G/T and G82S polymorphisms in the receptor for advanced glycation end products gene with diabetic retinopathy in Chinese population. AB - BACKGROUND: The receptor for advanced glycation end products (RAGE) plays an important role in the pathogenesis of diabetic complications. The aim of this study is to investigate the association of the 1704 G/T and G82S polymorphisms in the RAGE gene with diabetic retinopathy (DR). METHODS: The 1704G/T and G82S polymorphisms were genotyped in 340 Type 2 diabetes (T2DM) without DR subjects (DR- group); 166 T2DM with DR subjects (DR+ group), and 182 normal glucose tolerant subjects (NGT group). The genotypes were detected by the methods of ligase detection reaction coupled PCR. RESULTS: There was no evident difference in the 1704G/T and G82S genotypic and allelic frequencies distribution between NGT and T2DM subjects. However, the frequences of G/A+AA genotypes (60.6%) and A allele (36.4%) of G82S were significantly higher in DR+ group than those (38.4%; 20.9%, respectively) in DR- group (p=0.01and p=0.007, respectively). Furthermore, haplotype analysis revealed that the frequency of G-A haplotype containing 1704G and 82S allele in DR+ group was significantly higher than that in DR- group (33.5% vs 19.6%, p=0.01). The multiple logistic regression analysis indicated that the G82S polymorphism [odds ratio (OR): 2.964, 95% confidence interval (CI): 1.013-5.46, p=0.029] and diabetes duration (OR: 1.013, 95% CI: 1.007-1.02, p<0.001) were independent risk factors for DR. CONCLUSIONS: G82S polymorphism in the RAGE gene is associated with DR and G-A haplotype containing 1704G and 82S allele might be a genetic marker of DR in Chinese T2DM patients. PMID- 19542746 TI - Diabetes mellitus after kidney transplantation: role of the impaired fasting glucose in the outcome of kidney transplantation. AB - INTRODUCTION: Most patients suffering from chronic renal insufficiency show impaired carbohydrate metabolism. Our goals were to analyze the accumulated incidence of impaired fasting glucose (IFG) and post-transplant diabetes mellitus (PTDM) after kidney transplantation in our hospital, to assess their impacts on the survival of the graft and of the patient, and to discover the major risk factors for the development of PTDM. MATERIALS AND METHODS: We examined alterations in carbohydrate metabolism in 920 adult patients after they received kidney transplantation. Patients were followed for a minimum period of 5 yr. RESULTS: One year after transplantation, 12.8% of the patients had developed PTDM, and 10.3% showed an IFG level. The IFG had a negative and statistically significant influence on graft and patient survival. Host and donor age, weight, hepatitis C virus infection, and acute rejection were found to be significant risk factors. DISCUSSION: Our study found a high incidence of PTDM, as described in previous studies, but with an emphasis on a greater role played by IFG, not only in its incidence, but also as a prognostic factor for the outcome of graft and patient survival. Identifying patients at risk of developing PTDM is important in offering them early and appropriate treatment. PMID- 19542748 TI - Mutant epidermal growth factor receptor vIII increases cell motility and clonogenecity in a prostate cell line RWPE1. AB - Epidermal growth faxtor receptor (EGFR)-vIII mutant has been demonstrated to over express as prostatic neoplasms progressed from intraepithelial changes to metastatic disease. In this study, we transfected the EGFRvIII expression vector into an immortalized normal prostate epithelium cell line RWPE-1 and established stable transfectants. The cell growth, glandular morphogenesis, cell motility, and soft-agar colony formation efficiency were then studied. The results showed that EGFR-vIII mutation increased the RWPE1 cell motility and clone formation efficiency, while it had no significant effect on the cell growth when compared to non-transfected as well as mock transfected RWPE-1 cells. Moreover, EGFR-vIII changed the RWPE1 acinar morphogenesis. Further study showed that these effects of EGFR-vIII mutation may be related to down-regulation of E-cadherin and up regulation of beta-catenin. PMID- 19542747 TI - Thyrotropin variations may explain some positive radioiodine therapy scans in patients with negative diagnostic scans. AB - Thyroglobulin (Tg) is a specific marker of residual thyroid cancer or tumor recurrence. In patients with elevated Tg levels and negative diagnostic radioiodine (131I) whole-body scans (dWBS), administration of a therapy dose may reveal foci that were not initially apparent. The aim of this study was to identify factors, other than 131I activity, which might explain why a post therapy 131I whole-body scan is sometimes positive despite a negative dWBS. PATIENTS AND METHODS: We reviewed data on all patients with elevated Tg levels and negative dWBS with 185 MBq 131I off-T4 at followup, who subsequently received an empiric therapy dose of 3700 MBq of 131I. During a 5-yr period, 22 patients met these criteria. 131I therapy could be given immediately after negative dWBS in 9 patients, with an average of 8 extra days of hypothyroidism. In the other 13 patients, therapy was given an average of 8 months later. RESULTS: The therapy scan was negative in 16 patients, while it showed uptake in the thyroid bed in 5 patients and distant metastases in two. In the latter two patients, the TSH level was suboptimal at the time of dWBS (9 and 25 microIU/ml), and had risen to 34 and 70 microIU/ml respectively at the time of therapy. Overall, a positive scan following therapy occurred in 7 patients (6/9 patients treated immediately and 1/13 patients treated in a separate setting; p<0.01). In patients with positive therapy scans, the mean TSH level was 73 microIU/ml at the time of dWBS and 103.5 microIU/ml at the time of therapy (41% increase; p<0.05). In patients with negative therapy scans the mean TSH level was 84 microIU/ml at dWBS and 86 microIU/ml at the time of the therapy scan (2% increase). CONCLUSIONS: Our study suggests that interval increase in TSH level with a longer period of stimulation may have contributed to making the whole-body scan positive at the time of therapy. Nowadays, patients with elevated Tg are directly given a therapy dose of 131I. Special care should be taken when preparing patients who have been on suppressive levothyroxine therapy for a long time, in order to avoid misclassifying the tumor as non-functioning. PMID- 19542749 TI - The relationship between adiponectin, progesterone, and temperature across the menstrual cycle. AB - BACKGROUND: The mechanisms regulating adiponectin, a highly abundant adipokine produced by adipocytes, have not been fully elucidated. Adiponectin levels are significantly higher in women when compared to men, suggesting sex-hormone involvement in its regulation. Previously, we have observed an inverse association between adiponectin and basal body temperature in pregnant women. These findings suggest that states where progesterone and temperature fluctuate, such as the menstrual cycle, could be associated with fluctuating adiponectin levels. AIM: The aim of this study was to examine the relationship between adiponectin, progesterone, and temperature across the menstrual cycle. SUBJECTS AND METHODS: A prospective study was performed. Fifteen non-obese pre-menopausal female subjects, all with regular cycles, and on no medication recorded a daily temperature and underwent blood sampling, indirect calorimetry, and bio impendence studies in both the follicular and luteal phases of the menstrual cycle. RESULTS: Serum adiponectin levels did not vary significantly across the menstrual cycle or between those who did and did not ovulate. No correlation was found between adiponectin levels and sex steroids, insulin and glucose levels or basal energy expenditure and body composition. CONCLUSIONS: Our results indicate that adiponectin is not related to sex steroids or body composition in healthy premenstrual women. PMID- 19542750 TI - Somatotropin adenoma and resistance to thyroid hormone. AB - Resistance to thyroid hormone (RTH) is a rare disease characterized by non suppressed TSH in spite of high free thyroid hormone levels. Up to date, in the literature, there are more than 600 RTH cases, but co-incidental hypophyseal adenoma was reported in only 1 case. In the literature, despite reported cases with thyrotropinoma accompanying RTH, we could not find a case with somatotropinoma accompanying RTH. Here, we report a 34-yr-old male patient, who was admitted to the hospital with complaints of dyspnea, chest pain, and palpitation in 2003. His alpha- subunit value was normal and the alpha subunit/TSH molar ratio was <1. His response to TRH stimulation test was normal. His TSH level was suppressed in the T3 suppression test. Hypophyseal magnetic resonance imaging showed a 6-mm hypophyseal microadenoma. Levels of all anterior hypophyseal hormones, including GH and IGF-I, were normal. Oral glucose tolerance test (OGTT)-GH suppression test was normal. The patient was followed with the diagnosis of RTH and incidental hypophyseal adenoma. After 3 yr, because of high levels of IGF-I: 901 ng/ml (68-324), the OGTT-GH suppression test was reported and no suppression was detected. Thus, the patient was referred to surgery with the pre-diagnosis of RTH and acromegaly. Immunohistochemistry was showed as strong GH staining with low Ki 67 index while TSH and other anterior hypophyseal hormones stainings were negative. Post-operative thyroid hormones were still high. PMID- 19542752 TI - Elevations of inflammatory cytokines during and after pregnancy in gestational diabetes. PMID- 19542751 TI - Autoimmune hemolityc anemia concomitant with sequential autoimmune hepatitis primary biliary cirrhosis overlap syndrome and Hashimoto's thyroiditis: a new entity of autoimmune polyendocrine syndrome. PMID- 19542753 TI - High prevalence of vitamin D insufficiency during late winter and spring in healthy young women in Germany. PMID- 19542754 TI - Drawing of a tooth to a giant-acromegalic man: Indian bas-relief (II century BC). PMID- 19542755 TI - Graves' disease prevalence in a young population with Turner syndrome. PMID- 19542756 TI - Maternal isolated hypothyroxinemia: To treat or not to treat? PMID- 19542758 TI - Unusual endocrine and somatic phenotypic abnormalities in a 14-year-old boy with classic Klinefelter syndrome (47,XXY). PMID- 19542757 TI - Abnormal serum alanine aminotransferase levels are associated with impaired insulin sensitivity in young women with polycystic ovary syndrome. AB - BACKGROUND AND AIM: Non-alcoholic fatty liver disease (NAFLD) and polycystic ovary syndrome (PCOS) are both associated with insulin resistance.We assessed whether NAFLD is associated with impaired insulin sensitivity in PCOS women independently of age and total adiposity. SUBJECTS AND METHODS: We enrolled 14 young PCOS women with NAFLD, 14 women with PCOS alone and 14 healthy controls, who were matched for age, body mass index, and total body fat (by bio-impedance analyzer). NAFLD was diagnosed by the surrogate measure of abnormal serum alanine aminotransferase (ALT) concentrations (defined as ALT>19 U/l) after excluding other secondary causes of liver disease (alcohol, virus, and medications). Insulin sensitivity was measured by euglycemic hyperinsulinemic clamp. RESULTS: Insulin sensitivity was markedly decreased (p<0.001) in PCOS women with abnormal ALT levels, whereas it was similar between PCOS women with normal ALT levels andmatched healthy controls (8.3+/-2.5 vs 12.1+/-1.7 vs 13.2+/-1.8 mg/min x kg of fat-free mass, respectively). PCOS women with abnormal ALT levels also had higher plasma triglycerides and lower HDLcholesterol concentrations than those with PCOS alone. There was a strong inverse association between serum ALT levels and insulin sensitivity in the whole group of PCOS women (r=-0.59, p=0.0013). CONCLUSIONS: Abnormal serum ALT levels, as surrogate measure of NAFLD, are closely associated with impaired insulin sensitivity in young PCOS women in a manner that is independent from the contribution of age and total adiposity. Early recognition of NAFLD by radiological imaging tests in this group of young patients is warranted. PMID- 19542759 TI - Prospective evaluation of tumor size and hormonal status in adrenal incidentalomas. AB - BACKGROUND: Because of the increased use of imaging interventions, more subjects have been diagnosed with adrenal incidentaloma in recent years. AIM: To evaluate the risk of mass enlargement, hormone hypersecretion and development of adrenal carcinomas during short-term followup. SUBJECTS AND METHODS: There were 317 subjects with incidentally discovered adrenal tumors in the registry. Forty subjects were excluded because of clinically overt hormone secretion at diagnosis and subjects with complete data were included in radiological (no.=150) and hormonal (no.=150) follow- up. Radiological evaluation was performed with computed tomography (CT) and/or magnetic resonance imaging (MRI). There were 143 subjects with adrenal adenomas and 7 subjects with other tumor types (cyst or myelolipoma). Median follow-up duration was 24 months. RESULTS: Increase in tumor size was detected in 25 subjects (17.4%) with adenomas and 1 subject with adrenal myelolipoma (14.3%). Decrease in tumor size was found in 7 subjects (4.8%) with adrenal adenomas. One patient was diagnosed with adrenocortical carcinoma during follow-up. In subjects with non-functioning adrenal adenoma (NFA, no.=120) or subclinical Cushing syndrome (sCS) (no.=30), no subject developed clinically overt hormone hypersecretion, while 8 (6%) subjects in the NFA group developed sCS. Tumor diameter and follow-up duration were significantly higher in subjects who developed sCS. CONCLUSION: In conclusion, we demonstrated that, despite being infrequent, adrenal tumors may increase in size, develop overt or subclinical hormone secretion or feature malignant transformation. Therefore, radiological and hormonal follow-up should be recommended to the patients. More investigations are needed for the establishment of long-term follow-up protocols. PMID- 19542760 TI - Diet influences the content of bioactive peptides in goat milk. AB - Leptin, insulin, ghrelin, and IGF-I are circulating peptide hormones concerned with energy homeostasis and the regulation of GH axis. They are present in human milk, and are thought to promote neonatal development. The aim of the present study was to detect these substances in goat milk and determine whether their levels can be modified by changing the macronutrient content of the lactating animals' diet. Sixteen Saanen goats in mid-lactation were divided into two balanced groups, one given a diet containing 17% starch (LS) and the other a diet of 33% starch (HS). Eighty days later, leptin, insulin, ghrelin, and IGF-I were determined by human radioimmunoassay kits in plasma before and after feeding, and in sonicated milk centrifuged to remove fat from morning and evening milking. The HS diet was associated with higher plasma and milk insulin and IGF-I, and plasma ghrelin. Leptin, insulin, and ghrelin in milk were two-three times higher than in plasma; milk IGF-I was only 5-20% of plasma level. Plasma insulin correlated positively with plasma IGF-I; morning milk IGFI and insulin correlated positively with morning plasma levels. These findings demonstrate that human immuno activities of bioactive peptides are present in goat milk, and also that levels of insulin and IGF-I in milk can be altered by changing the macronutrient content of the diet. Further research is required to determine whether these substances can be transferred from the milk to suckling animals and humans, and whether they have biological activity in such animals. PMID- 19542762 TI - Chest pain relieved with a bronchodilator or other asthma drugs. AB - BACKGROUND: Although many patients who experience chest pain or pressure consult their physicians, unfortunately a large number of them do not, and consequently they remain undiagnosed and untreated. Chest pain, in a subset of these patients, may be relieved with a bronchodilator or other asthma drugs. METHODS: This retrospective study included twenty cases of chest pain that were relieved with asthma drugs. Chest pain was categorized into three types: chest pain variant asthma, bronchial asthma with chest pain, and non-asthmatic allergic chest pain. Chest pain variant asthma was defined as chest pressure that improved in response to a bronchodilator, without the characteristic attacks of bronchial asthma. Bronchial asthma with chest pain was defined as chest pressure, with the characteristic attacks of bronchial asthma that improved following the administration of a leukotriene receptor antagonist, systemic corticosteroid, or bronchodilator. Non-asthmatic allergic chest pain was defined as chest pressure without the typical asthma attack, but with chest pressure that improved in response to a leukotriene receptor antagonist or systemic corticosteroid, but not a bronchodilator. RESULTS: Fourteen cases of chest pain were diagnosed as variant asthma, three cases were diagnosed as bronchial asthma with chest pain, and three cases were diagnosed as non-asthmatic allergic chest pain. CONCLUSIONS: The results suggest that the mechanism underlying chest pain that is relieved with asthma drugs can involve either an airway constriction pathway or a non constrictive pathway presumably airway inflammation. Analysis of the patient's response to treatment with asthma medication is useful for the correct diagnosis of the source of chest pain. PMID- 19542761 TI - Endogenous and exogenous thioredoxin 1 prevents goblet cell hyperplasia in a chronic antigen exposure asthma model. AB - BACKGROUND: Goblet cell hyperplasia with mucus hypersecretion contribute to increased morbidity and mortality in bronchial asthma. We have reported that thioredoxin 1 (TRX1), a redox (reduction/oxidation)-active protein acting as a strong antioxidant, inhibits pulmonary eosinophilic inflammation and production of chemokines and Th2 cytokines in the lungs, thus decreasing airway hyperresponsiveness (AHR) and airway remodeling in mouse asthma models. In the present study, we investigated whether endogenous or exogenous TRX1 inhibits goblet cell hyperplasia in a mouse asthma model involving chronic exposure to antigen. METHODS: We used wild-type Balb/c mice and Balb/c background human TRX1 transgenic mice constitutively overproducing human TRX1 protein in the lungs. Mice were sensitized 7 times (days 0 to 12) and then challenged 9 times with ovalbumin (OVA) (days 19 to 45). Every second day from days 18 to 44 (14 times) or days 35 to 45 (6 times), Balb/c mice were treated with 40 microg recombinant human TRX1 (rhTRX1) protein. Goblet cells in the lungs were examined quantitatively on day 34 or 45. RESULTS: Goblet cell hyperplasia was significantly prevented in TRX1-transgenic mice in comparison with TRX1 transgene negative mice. rhTRX1 administration during OVA challenge (days 18 to 44) significantly inhibited goblet cell hyperplasia in OVA-sensitized and -challenged wild-type mice. Moreover, rhTRX1 administration after the establishment of goblet cell hyperplasia (days 35 to 45) also significantly ameliorated goblet cell hyperplasia in OVA-sensitized and -challenged wild-type mice. CONCLUSIONS: Our results suggest that TRX1 prevents the development of goblet cell hyperplasia, and also ameliorates established goblet cell hyperplasia. PMID- 19542763 TI - Very short gastroesophageal acid reflux during the upright position could be associated with asthma in children. AB - BACKGROUND: Gastroesophageal reflux disease (GERD) is diagnosed by the reflux index of 24-hour pH monitoring (pH monitoring). In our previous study, GER episodes during the upright position were more frequent than those during the supine position in asthmatic children. In this study, we investigated the clinical usefulness of the mean hourly number of acid refluxes, designated as the mean number of acid refluxes/hour (h) during the upright position in addition to the pH index for the diagnosis of GERD. METHODS: The subjects were 22 preschool asthmatic children. When the reflux index was over 4% or the mean number of acid refluxes/h during the upright position were three times more frequent than those during the supine position even if the reflux index was below 4%, we prescribed famotidine. Children whose asthmatic symptoms improved with famotidine were included in a GERD group. Children who did not meet the criteria by pH monitoring were included in a non-GERD group in asthmatic children. RESULTS: The GERD group was comprised of 9 children. In 2 out of 9 GERD group children, the reflux index was below 4%. The median of the mean number of acid refluxes/h during the upright position was 12.9 in the GERD group, and 3.15 in the non-GERD group. The mean number of acid refluxes/h during the upright position were associated with asthmatic symptoms (p < 0.05). CONCLUSIONS: Reflux during the upright position was associated with asthmatic symptoms. The mean number of acid refluxes/h during the upright position in addition to the reflux index could be useful in the diagnosis of GERD when associated with asthma. PMID- 19542765 TI - Effect of suplatast tosilate on antileukotriene non-responders with mild-to moderate persistent asthma. AB - BACKGROUND: Immunomodulatory therapy has been recently introduced for the management of asthma. Suplatast tosilate (ST), a new immune-modifying drug, is known to improve the airway function by inhibiting the release of Th-2 cytokines. However, its efficacy as a controller listed in the guideline, Global Initiative for Asthma 2005 has not been established. In this study we investigated the role of ST in leukotriene receptor antagonist (LTRA) non-responders with mild-to moderate persistent asthma before initiating corticosteroids inhalation therapy. METHODS: This was a prospective open-level clinical trial. LTRAs was given to 41 patients with asthma for 4 weeks and clinical efficacy was assessed using daily symptom scores. The 10 patients, aged 2.5-8.5 years, who failed to show clinical improvement, were defined as LTRA non-responders. After a 1-week washout period, the efficacy of ST was investigated and compared with LTRA non-responders for the following 4 weeks. RESULTS: LTRA non-responders showed a significant improvement in the average symptom score, peak expiratory flow, use of rescue medication and the proportion of symptom-free days with ST therapy. CONCLUSIONS: ST is a good choice for patients who have failed to respond to LTRAs. ST should therefore be added to the list of treatment options for such patients. PMID- 19542764 TI - Polyclonal IgE induces mast cell survival and cytokine production. AB - BACKGROUND: Ag-dependent activation of IgE-bearing mast cells is a critical first step in immediate hypersensitivity and other allergic responses. Recent studies have revealed Ag-independent effects of monoclonal mouse IgE molecules on mast cell survival and activation. However, no studies have been performed on the effects of polyclonal IgE molecules. Here, we tested whether polyclonal mouse and human IgE molecules affect survival and cytokine production in mast cells. METHODS: Mast cells were cultured in the presence of polyclonal mouse and human IgE molecules, and cell survival and cytokine production were analyzed. RESULTS: Polyclonal mouse IgE molecules in sera from mice with atopic dermatitis-like allergic skin inflammation, enhanced survival and cytokine production in mast cell cultures. Similar to the effects of monoclonal IgE, the polyclonal IgE effects were mediated by the high-affinity IgE receptor, FcepsilonRI. Human polyclonal IgE molecules present in sera from atopic dermatitis patients were also capable of activating mast cells, and inducing IL-8 production in human cord blood-derived mast cells. CONCLUSIONS: These results imply that polyclonal IgE in atopic dermatitis and other atopic conditions might modulate mast cell number and function, thus amplifying the allergic response. PMID- 19542766 TI - The efficacy of early treatment of seasonal allergic rhinitis with benifuuki green tea containing O-methylated catechin before pollen exposure: an open randomized study. AB - BACKGROUND: We previously reported that 'benifuuki' green tea containing O methylated catechin significantly relieved the symptoms of perennial or seasonal rhinitis compared with a placebo green tea that did not contain O-methylated catechin in randomized double-blind clinical trials. In this study we assessed the effects of 'benifuuki' green tea on clinical symptoms of seasonal allergic rhinitis. METHODS: An open-label, single-dose, randomized, parallel-group study was performed on 38 subjects with Japanese cedar pollinosis. The subjects were randomly assigned to long-term (December 27, 2006-April 8, 2007, 1.5 months before pollen exposure) or short-term (February 15, 2007: after cedar pollen dispersal--April 8, 2007) drinking of a 'benifuuki' tea drink containing 34 mg O methylated catechin per day. Each subject recorded their daily symptom scores in a diary. The primary efficacy variable was the mean weekly nasal symptom medication score during the study period. RESULTS: The nasal symptom medication score in the long-term intake group was significantly lower than that of the short-term intake group at the peak of pollen dispersal. The symptom scores for throat pain, nose-blowing, tears, and hindrance to activities of daily living were significantly better in the long-term group than the short-term group. In particular, the differences in the symptom scores for throat pain and nose blowing between the 2 groups were marked. CONCLUSIONS: We conclude that drinking 'benifuuki' tea for 1.5 months prior to the cedar pollen season is effective in reducing symptom scores for Japanese cedar pollinosis. PMID- 19542767 TI - Two thirds of forest walkers with Japanese cedar pollinosis visit forests even during the pollen season. AB - BACKGROUND: The most common type of pollinosis in Japan is Japanese cedar pollinosis (JCP). While forest walking is a common form of recreation for Japanese people, it has been unclear whether forest walkers with JCP still choose to visit forested areas during the pollen season or whether they avoid those areas, and as such, the aim of this study was to investigate this question. METHODS: The study participants were all healthy men and women volunteers aged 20 years or over who visited the Tokyo University Forest in Chiba during 4 different days. The survey was conducted using self-administered questionnaires. RESULTS: The number of available responses was 498. Of these, 112 participants who experienced JCP were included in the analysis. Seventy-three participants (65.2%) responded that they visit forests even during the pollen season. The association between forest walking choices during the pollen season and self-rated levels of pollinosis symptoms was not statistically significant (Cramer's V = 0.13, p = 0.47). As many as 60% of the participants who reported serious symptom levels responded that they visit forested areas even during the pollen season. CONCLUSIONS: These results revealed that two thirds of forest walkers who had experienced JCP visited forests even during the pollen season. This indicates the further need for public service announcements informing people with JCP that the risk of pollen exposure and subsequent JCP reaction is increased by visiting forested areas during the pollen season. PMID- 19542768 TI - Neonatal nurse practitioners: a value to nursing. PMID- 19542769 TI - Advanced practice registered nurse consensus model. PMID- 19542771 TI - Pain management during retinopathy of prematurity eye examinations: a systematic review. AB - Retinopathy of prematurity (ROP) remains a concern for many preterm infants. Early detection and timely treatment have been shown to be effective in improving visual outcomes; moreover, it is crucial that a series of indirect ophthalmic examinations be performed until an infant is considered no longer at risk for the disease. The purpose of this systematic review is to summarize and evaluate the published evidence regarding characteristics and effectiveness of pain management interventions during the ROP examination. Implications for practice are discussed and suggestions for further research are made. Despite the general consensus that ROP examination is a painful procedure with considerable amount of discomfort, evidence shows that pain management during the ROP examination is inadequate. Although there are currently clear recommendations and guidelines for performing the ROP examination, there are no standard protocols for pharmacological and nonpharmacological pain management during the ROP examination. This is an area where much work is still needed to address the needs of the infant during this critical examination. PMID- 19542772 TI - Rapid sequence intubation in the neonate. AB - Rapid sequence intubation (RSI) is premedication prior to intubation that includes atropine, a sedative, and a neuromuscular blockage. Rapid sequence intubation is infrequently performed in neonates despite evidence that it is safe and effective. Neonates that experience endotracheal intubation often display apnea and cardiac arrhythmias, decreased or obstructed nasal airflow, increased systolic blood pressure, and decreased heart rate and transcutaneous oxygen tension. Infants can also experience increased anterior fontanel pressure, which can place them at greater risk for intraventricular hemorrhage. Rapid sequence intubation has been shown to facilitate better intubation conditions including no movement from the infant and better visualization of the airway. Infants receiving RSI were successfully intubated twice as fast as infants who were not premedicated. Infants with premedication also had fewer changes in baseline heart rate. Neonatal RSI can be easily and safely performed in the neonate. Knowledge and skill allow for the best conditions when intubating the infant. Future research must focus on the best combination of medications for RSI in the neonate. PMID- 19542774 TI - Klippel-Trenaunay syndrome: a case study. AB - Klippel-Trenaunay syndrome is a rare but well-documented congenital malformation. Klippel-Trenaunay syndrome has sometimes been used interchangeably with Klippel Trenaunay-Weber syndrome. However, Klippel-Trenaunay syndrome is the correct term used for the triad of congenital anomalies. Klippel-Trenaunay-Weber syndrome or Parkes-Weber syndrome is accepted as a separate entity consisting of the triad of Klippel-Trenaunay syndrome accompanied by a clinically apparent arteriovenous fistula. Hemodynamically insignificant arteriovenous malformations do not preclude a diagnosis of Klippel-Trenaunay syndrome. It is important to differentiate between the 2 syndromes because treatment and prognosis are so different. Parkes-Weber syndrome has a poor prognosis for limb viability. This article describes a case study of an infant presenting with Klippel-Trenaunay including a review of the syndrome and treatment recommendations. PMID- 19542775 TI - Neonatal nurse practitioners: past, present, and future. AB - The advanced practice nursing role in neonatal intensive care enjoys an almost 40 year history. The 1970s and the 1980s were fraught with growing pains, including what to call these providers, and role clarity that was defined in most settings by community need. With the birth of NANN, 25 years ago, the neonatal nurse practitioner (NNP) role began to receive support and advocacy that led to the development of educational standards and definition of the role. This article reviews the role's history and identifies current and future issues that will require attention by the national NNP leadership. PMID- 19542776 TI - Lansoprazole, ranitidine, and metoclopramide: comparison of practice patterns at 4 level III NICUs within one healthcare system. AB - PURPOSE: Evidence is needed to guide NICU use of lansoprazole (Prevacid), ranitidine (Zantac), and metoclopramide (Reglan). As a step toward that goal, we conducted a historic cohort analysis of all patients who received any of these medications in 4 Intermountain Healthcare NICUs during the year 2006. SUBJECTS: Data were obtained from all patients admitted between January 1 and December 31, 2006, to any of 4 Intermountain Healthcare NICUs. DESIGN: This was a retrospective descriptive design. METHODS: Data were obtained from electronic pharmacy records and electronic medical records. The NICUs involved were blinded and included McKay-Dee Hospital Center, Ogden, Utah; LDS Hospital, Salt Lake City, Utah; Utah Valley Regional Medical Center, Provo, Utah; and Dixie Regional Medical Center, St George, Utah. RESULTS: Although the demographics of the patients at the 4 centers were similar, significant differences were seen among the centers in drug use patterns. Lansoprazole use ranged from a high of 17% of patients in one center to a low of 7% of patients in another. Ranitidine use ranged from 9% in one center to 1% in another. Metoclopramide use ranged from 9% of patients in one center to <1% in another. CONCLUSIONS: The extreme variability among the centers in use patterns of these 3 medications suggests lack of an adequate evidence base to guide practice and indicates that case controlled studies or random controlled trials are needed to devise a consistent evidence based approach. PMID- 19542777 TI - Benevolent injustice: a neonatal dilemma. AB - There is a little-recognized cohort of NICU patients whose outcomes are the result of a "benevolent injustice" in their healthcare course. Many of these infants are saved by technology; however, they are left both medically fragile and medically dependent, and many of them are required to live in a medical facility. Many of these babies never get to go home with their parents. This emerging cohort of patients may evolve from the difficult ability to prognosticate outcomes for neonates, overtreatment, and acquiescing to parental demands for continued aggressive care. Neonatology is an unpredictable process and one that is never intended to harm, but carries with it the potential of devastating consequences, thus creating a benevolent injustice. PMID- 19542778 TI - Mobility, assistive technology use, and social integration among adults with spina bifida. AB - OBJECTIVE: Many individuals with spina bifida have impairments that limit mobility and functional independence. Sedentary lifestyles and social isolation are very prevalent. This study evaluated the association between the use of mobility devices and degree of socialization. DESIGN: A retrospective chart review was performed on 208 adults with spina bifida attending a university-based clinic. Data collected included the Craig Handicap Assessment Reporting Technique Short Form, Beck Depression Inventory, and data on wheelchair and other assistive technology use. We hypothesized that community and home mobility and social integration, as measured by the Craig Handicap Assessment Reporting Technique Short Form, would be lower for manual and power wheelchair users than for ambulators, regardless of depression scores or shunt history. RESULTS: We found that individuals with spina bifida who used both manual and power wheelchairs do have lower daily home and community activity levels compared with ambulators, but that most individuals with spina bifida have low social integration and economic self-sufficiency scores, regardless of whether they can ambulate or use wheelchairs. These findings were not explained by wheelchair quality because most were prescribed high-quality devices. A high prevalence of depression was also found. CONCLUSIONS: Special considerations for wheelchair provision are discussed. Additional research is needed to identify other barriers to social integration. PMID- 19542779 TI - Predicting recovery after fibular nerve injury: which electrodiagnostic features are most useful? AB - OBJECTIVE: Although it is important to have strong predictors of outcome in peripheral mononeuropathies so that appropriate management can be instituted early, little is known about the prognostic value of electrodiagnostic results for these lesions. Our objective in this study was to evaluate the prognostic value of electrodiagnostic studies in fibular neuropathy. DESIGN: In this retrospective study, 39 of 138 subjects with fibular neuropathy met the inclusion criteria. Electrodiagnostic results at the time of testing were evaluated for their value in predicting outcome. Good outcome was defined as grade 4 or higher strength on the Medical Research Council Scale in ankle dorsiflexion. RESULTS: Compound muscle action potential responses from extensor digitorum brevis and tibialis anterior predicted prognosis: 81% of subjects with any tibialis anterior response and 94% with any extensor digitorum brevis response had a good outcome vs. those with absent responses (46% and 52%, respectively). Importantly, there was still a high likelihood of good outcome with absent compound muscle action potential responses. Tibialis anterior compound muscle action potential gave additional prognostic information when extensor digitorum brevis response was absent. Recruitment in tibialis anterior was predictive in traumatic cases. All patients with nontraumatic compression had a good outcome. CONCLUSIONS: We conclude that electrodiagnostic studies produce useful prognostic information in fibular neuropathy, particularly in traumatic cases. PMID- 19542780 TI - Influence of glove type on mobility performance for wheelchair rugby players. AB - OBJECTIVE: The purpose of this study was to determine the effectiveness of different glove types on mobility performance in a series of field tests specific to wheelchair rugby. DESIGN: Ten international wheelchair rugby players performed three drills in each glove condition: (i) players' current glove selection, (ii) American football glove, (iii) building glove, and (iv) new prototype glove. Performance was assessed by a combination of outcomes including test times, peak velocities, and accelerations within each drill. Peak velocities and accelerations were measured using a velocometer sampling at 100 Hz. RESULTS: A two-way analysis of variance with repeated measures revealed that participants performed statistically better for measures of acceleration and sprinting when wearing their current choice of glove compared with the new prototype glove (P < 0.05). Subjective data identified that players also favored their current gloves compared with others, although slight discrepancies were evident among classification levels concerning the building gloves. Building gloves seemed to be less suited to low-point players because of the reduced grip and protection they provided. CONCLUSIONS: It may be concluded that participants' current gloves that have been modified for the specific demands of wheelchair rugby are more effective for aspects of mobility performance than other glove types. PMID- 19542781 TI - The 200-m fast-walk test compared with the 6-min walk test and the maximal cardiopulmonary test: a pilot study. AB - OBJECTIVE: The 200-m fast-walk test has been proposed as a high- intensity performance test in healthy, elderly subjects. Adaptation of low-risk coronary artery disease patients during this test were compared with those in a 6-min walk test and a maximal cardiopulmonary exercise test. DESIGN: Thirty patients with stable coronary artery disease (51.9 +/- 8.7 yrs), referred to the cardiac rehabilitation department, performed a cardiopulmonary exercise test, then a 200 m fast-walk test and a 6-min walk test in a random order, before and after the training period (6 wks, 3 days per week). Heart rate was monitored during each test. Peak workload of cardiopulmonary exercise test, distance walked on the 6 min walk test, and time to perform the 200-m fast-walk test were measured. A subsample of ten patients performed the exercise test with gas exchange measurements, with ventilatory threshold determination. RESULTS: All subjects completed walk tests without complaint or incidents. Compared with the cardiopulmonary exercise test, the cardiac relative intensity was higher during the 200-m fast-walk test than during the 6-min walk test, both before (89.6% vs. 78.1% of cardiopulmonary exercise test maximal heart rate; P < 0.05) and after (83.8% vs. 74.3%; P < 0.05) training. Among the subsample of ten patients, the 200-m fast-walk test heart rate was significantly higher than the ventilatory threshold heart rate, which did not differ from the 6-min walk test heart rate. The 200-m fast-walk test time significantly decreased after training (-9.1%, P < 0.01). CONCLUSION: In patients with coronary artery disease at low risk, the 200 m fast-walk test explores higher levels of cardiorespiratory capacity than the 6 min walk test. Thus, this could be a useful field test in complement to the cardiopulmonary exercise test to assess functional capacity improvement and update training targets regularly during the course of high-intensity rehabilitation programs in this population. PMID- 19542782 TI - Validation of functional fitness tests as surrogates for strength measurement in frail, older adults with chronic obstructive pulmonary disease. AB - OBJECTIVE: This study evaluated the relationship of functional field tests to traditional laboratory strength tests in 40 frail, older adults with chronic obstructive pulmonary disease. DESIGN: Participants completed two trials of upper (chest press) and lower (leg press) body maximal strength (maximal weight lifted one time) testing and one trial of functional fitness testing (Arm Curl, Lift and Reach, Chair Stand, Up and Go). The maximal weight lifted one-time values from both trials were compared with functional fitness test values to identify valid surrogates. RESULTS: Among upper-body functional fitness tests, the Arm Curl had a moderately strong relationship to chest press (trial 1: r = 0.55, P = 0.01; trial 2: r = 0.56, P = 0.01) whereas the Lift and Reach did not. Among lower-body functional fitness tests, the Chair Stand had a fairly strong relationship to leg press (trial 1: r = 0.46, P = 0.01; trial 2: r = 0.38, P = 0.05), but the Up and Go did not. CONCLUSIONS: Only the Arm Curl and Chair Stand tests were valid surrogates. Although multiple field tests to measure strength in a clinical setting may be desirable, these data support limiting functional testing to the Arm Curl for upper-body and the Chair Stand for lower-body strength assessment. PMID- 19542785 TI - Re: pre- and postcardiopulmonary rehabilitation in hospitalized patients undergoing coronary artery bypass surgery: a randomized control trial. PMID- 19542788 TI - Re: cutaneous electrical stimulation treatment in unresolved facial nerve paralysis. PMID- 19542789 TI - The role of the physiatrist in stroke rehabilitation: a European survey. PMID- 19542790 TI - Acute care and rehabilitation of patients with stroke in Hungary. PMID- 19542791 TI - Predictive factors of postoperative pain after day-case surgery. AB - OBJECTIVES: Despite the growing number of ambulatory operations knowledge of predictive factors of postoperative pain after ambulatory surgery is limited. Therefore, the aim of this study was to identify predictive factors of postoperative pain after ambulatory surgery. METHODS: In this cross-sectional study, 648 patients were included. A wide variety of elective ambulatory operations were performed. Pain assessments were made before the operation and during a 4-day period postoperatively, using a 100 mm visual analog scale. Patient characteristics, type of surgery, and type of anesthesia were recorded. In addition, preoperative expectations of postoperative pain by physician and patient were assessed. Finally, several scores about psychologic parameters were measured: pain catastrophizing, surgical anxiety, and optimism. Stepwise logistic regression analysis was performed to identify factors that independently predict the risk of having postoperative pain (defined by a visual analog scale >40 mm) on days 0 to 4. RESULTS: The most important predictor of postoperative pain was the presence of preoperative pain. Other predictors were anticipated postoperative pain by the clinician, preoperative high expectations of postoperative pain by the patient, younger age, and fear of short-term consequences of the operation. Regional anesthetic technique compared with general anesthesia decreased the risk of acute postoperative pain only on the day of the operation. DISCUSSION: Several predictive factors of postoperative pain after ambulatory surgery were identified in this study. These factors should be taken into account when planning postoperative analgesia for ambulatory surgery. PMID- 19542792 TI - Comparisons of the efficacy and safety of duloxetine for the treatment of fibromyalgia in patients with versus without major depressive disorder. AB - OBJECTIVES: To investigate whether comorbid major depressive disorder (MDD) influenced the efficacy and safety of duloxetine in treating fibromyalgia (FM). METHODS: This was a post-hoc analysis using pooled data from 4 double-blind, placebo-controlled studies of patients with American College of Rheumatology defined primary FM with or without MDD. Patients were randomized to duloxetine [60 or 120 mg/d (N=797)] or placebo (N=535) for approximately 3 months. Efficacy measures included the Brief Pain Inventory average pain score, 17-item Hamilton Depression Rating Scale, Fibromyalgia Impact Questionnaire, and Patient's/Clinician's Global Impressions of Improvement/Severity scales. RESULTS: At baseline, 26% of patients met diagnostic criteria for MDD. At endpoint (3 mo or last observation), duloxetine showed significantly (P<0.05) greater improvement versus placebo on the Brief Pain Inventory, Fibromyalgia Impact Questionnaire, Patient's Global Impressions of Improvement scale, and Clinician's Global Impressions of Severity scale in patients with and without comorbid MDD. The effect of duloxetine on these efficacy measures was consistent across FM patients with or without MDD (P>0.1 for treatment-by-strata interaction). On the 17-item Hamilton Depression Rating Scale, duloxetine showed significantly (P<0.05) greater improvement versus placebo in patients with comorbid MDD. The safety profile of duloxetine versus placebo with respect to serious adverse events and discontinuation owing to adverse events was similar for FM patients with versus without MDD (P>0.1 treatment-by-strata interaction). DISCUSSION: Duloxetine was effective in reducing pain and other symptoms in FM patients with and without MDD and demonstrated a similar safety profile for both groups. PMID- 19542793 TI - Predictors of placebo response in pooled lamotrigine neuropathic pain clinical trials. AB - OBJECTIVES: One limitation of neuropathic pain clinical trials is the often large and variable extent of response in the placebo group, possibly obscuring true medication effects. We pooled data from 252 individuals in the placebo arms of 3 clinical trials of lamotrigine in patients with neuropathic pain to examine the relationship of baseline patient and study site characteristics with 12-week change in the Pain Intensity Numerical Rating Scale score (DeltaPI-NRS). The 574 patients in the pooled lamotrigine treatment arms were used as a replication dataset. MATERIALS AND METHODS: We performed univariable and multivariable regression analysis of predictors of DeltaPI-NRS. Clinical factors examined were baseline pain intensity score (mean daily PI-NRS over the week prior to randomization), age, sex, diagnosis, prior and concurrent gabapentin use, prior and concurrent tricyclic antidepressant use, pain duration, variability of daily pain scores during the baseline week, and slope of daily pain scores over the baseline week. Site factors evaluated were study site, US geographic region, recruitment rate, and recruitment period. RESULTS: Baseline PI-NRS and site recruitment rate were independent predictors of the 12-week DeltaPI-NRS in the last observation carried forward, observed case, and repeated measures analyses. Patients with higher baseline PI-NRS scores had a significantly greater 12-week reduction in pain intensity than patients with lower baseline scores. Patients within sites with a faster recruitment rate also had a significantly greater reduction of pain intensity than those in sites with slower recruitment. DISCUSSION: These results suggest that both patient and study site characteristics can influence the response in the placebo arms of neuropathic pain studies. PMID- 19542795 TI - Opioid-sparing effects of ketorolac and its correlation with the recovery of postoperative bowel function in colorectal surgery patients: a prospective randomized double-blinded study. AB - OBJECTIVES: Postoperative ileus (PI) is one of many common complications in major abdominal surgery. PI results in patient discomfort, increased gastrointestinal leakage, prolonged hospital stay, and increased medical expenses. In this study, we have investigated the morphine-sparing effects of ketorolac and its correlation with the duration of PI in patients with colorectal surgeries. METHODS: We collected data from 102 patients who had received elective colorectal resection. The patients were randomly allocated into 2 groups and received intravenous patient-controlled analgesia (IVPCA) morphine (M group) or IVPCA morphine plus ketorolac (M+K group). Time-scale morphine consumption (per 12 h), recovery of bowel functions (the first bowel movement and passage of flatus), pain scores, and opioid-related side effects were then recorded. RESULTS: Patients in the M+K group received 18.3% less morphine than those in the M group within 72 postoperative hours. The maximal opioid-sparing effects of ketorolac appeared in 12 to 24 postoperative hours. The onset of the first bowel movement and passage of flatus was significantly less in the M+K group than in the M group. The M group showed a 5.25 times greater risk of inducing PI, a result comparable with the M+K group in colorectal surgery patients. DISCUSSION: The addition of ketorolac to IVPCA morphine has demonstrated a clear opioid-sparing effect and benefits in regards to the shortening of the duration of bowel immobility. We suggest that adding ketorolac to morphine IVPCA be included in the multimodal postoperative rehabilitation program for the early restoration of normal bowel function. PMID- 19542794 TI - Prescription opioid aberrant behaviors: a pilot study of sex differences. AB - OBJECTIVES: Patients who are prescribed opioids often display 1 or more aberrant prescription use behaviors (eg, requesting early refills, borrowing medication from family), which raise concern among healthcare professionals. Little is known about the sex differences in specific types of aberrant behaviors or sex-specific predictors of such behaviors. The current study is aimed to begin addressing this gap in the literature. METHODS: A battery of anonymous, self-report assessments was administered to 121 (49 men, 72 women) chronic pain patients enrolled in an outpatient pain management clinic. Most of the participants were white women with an average age of 51.6 years (SD=13.2). RESULTS: Significantly more men than women were taking a prescribed opioid (91.7% vs. 77.8%, P=0.05). Women were significantly more likely than men to hoard unused medication (67.6% vs. 47.7%, P=0.04) and to use additional medications to enhance the effectiveness of pain medication (38.8% vs. 20.0%, P=0.04). A trend toward men using alternative routes of administration (eg, crushing and snorting pills) more often than women was observed (8.9% vs. 1.5%, P=0.08). Among men, high rates of aberrant prescription use behaviors were associated with current alcohol use and the use of oxycodone and morphine. Among women, use of hydrocodone was associated with high rates of aberrant prescription use behaviors. DISCUSSION: Some aberrant prescription use behaviors are common among chronic pain patients and may be sex-specific. Predictors of aberrant prescription use behaviors may also differ by sex. Additional research is needed to help identify aberrant prescription use behaviors that best predict sex-specific risk for developing opioid abuse or dependence. PMID- 19542797 TI - Efficacy of microcurrent therapy in the treatment of chronic nonspecific back pain: a pilot study. AB - OBJECTIVES: Microcurrent therapy (MCT) is a novel treatment for pain syndromes. The MCT patch is hypothesized to produce stimuli that promote tissue healing by facilitating physiologic currents. Solid evidence from randomized clinical trials is lacking. To evaluate the efficacy of MCT in treating aspecific, chronic low back pain, we conducted a double-blind, randomized, crossover, pilot trial. METHODS: Ten succeeding patients presenting with nonspecific, chronic low-back pain in our university hospital were included. Patients started with two, 9-day baseline period followed by a 5-day treatment periods. During the treatment periods, either a placebo or MCT (verum) patch was randomly assigned. Mean and worst pain scores were evaluated daily by a visual analog scale (VAS). Furthermore, analgesic use, side effects, and quality of life were assessed after each period. Differences between the last 4 days of a treatment period and the baseline period were calculated. Differences between verum and placebo periods per patient were compared using paired t tests. A 20-mm VAS score reduction was considered clinically relevant. RESULTS: The VAS score was lower during verum treatment, with a reduction [95% confidence interval (CI] of -0.43 (-1.74; 0.89) in mean and -1.07 (-2.85; 0.71) in worst pain. Analgesic use decreased during verum treatment, except for nonsteroid anti-inflammatory drug use, which increased. Quality of life improved during verum treatment. However, note of the findings were statistically significant. DISCUSSION: A positive trend in MCT use for aspecific, chronic low-back pain is reported. Further investigations are required to evaluate the significance and relevance of this. PMID- 19542796 TI - A randomized controlled trial of vapocoolant for pediatric immunization distress relief. AB - OBJECTIVES: The purpose of this study was to evaluate the effectiveness of vapocoolant for preschoolers' immunization injection pain relief. METHODS: Fifty seven 4 to 6-year-old children were randomized into vapocoolant alone or typical care conditions. Pain was measured at the baseline and at injection via self report, caregiver report, nurse report, and by an observational scale. RESULTS: Self-report of pain suggested that children in the vapocoolant alone condition demonstrated stronger increases in pain from baseline to injection than children in the typical care condition. All other measures showed significant increases in pain from baseline to injection, but no other measures indicated treatment effects. DISCUSSION: This study revealed that vapocoolant is not an effective pain management intervention for children's intramuscular injections. PMID- 19542798 TI - Quantification of abdominal wall pain using pain pressure threshold algometry in patients with chronic pelvic pain. AB - OBJECTIVE: Chronic pelvic pain (CPP) is a syndrome involving 1 or more pain generating organs in the pelvis, which includes pain from the lower anterior abdominal wall. This entity has been termed myofascial pain syndrome (MFPS), but its characteristics, definition, and quantification have not been well described. In this study, pain pressure threshold (PPT) testing of the lower anterior abdominal wall in CPP patients was performed to determine the range and distribution of values at each site, and the clinical utility of using PPT in a definition of MFPS. METHODS: Fifty-six patients evaluated in a CPP specialty clinic underwent PPT algometry of 14 sites on the lower anterior abdominal wall. These values were described and evaluated before and after treatment. PPT values were also evaluated in patients found to be drug seeking. RESULTS: Twenty percent of the PPT tests reached the threshold of 3 kgf/cm2. The abnormal tests usually formed a normal distribution. PPT testing had a weak but appropriate correlation with other pain symptom measures. After trigger point injection there was a 75% improvement in PPT, and response to medical therapy resulted in a 60% improvement. A composite measure was able to distinguish drug-seeking patients with statistical accuracy. DISCUSSION: PPT testing can be used to evaluate MFPS in CPP patients. One suggested definition would exclude patients with low scores in the upper abdomen while including patients with low scores in the lower abdomen. PMID- 19542799 TI - Multiple active myofascial trigger points and pressure pain sensitivity maps in the temporalis muscle are related in women with chronic tension type headache. AB - OBJECTIVE: To describe the common locations of active trigger points (TrPs) in the temporalis muscle and their referred pain patterns in chronic tension type headache (CTTH), and to determine if pressure sensitivity maps of this muscle can be used to describe the spatial distribution of active TrPs. METHODS: Forty women with CTTH were included. An electronic pressure algometer was used to assess pressure pain thresholds (PPT) from 9 points over each temporalis muscle: 3 points in the anterior, medial and posterior part, respectively. Both muscles were examined for the presence of active TrPs over each of the 9 points. The referred pain pattern of each active TrP was assessed. RESULTS: Two-way analysis of variance detected significant differences in mean PPT levels between the measurement points (F=30.3; P<0.001), but not between sides (F=2.1; P=0.2). PPT scores decreased from the posterior to the anterior column (P<0.001). No differences were found in the number of active TrPs (F=0.3; P=0.9) between the dominant side the nondominant side. Significant differences were found in the distribution of the active TrPs (chi2=12.2; P<0.001): active TrPs were mostly found in the anterior column and in the middle of the muscle belly. The analysis of variance did not detect significant differences in the referred pain pattern between active TrPs (F=1.1, P=0.4). The topographical pressure pain sensitivity maps showed the distinct distribution of the TrPs indicated by locations with low PPTs. CONCLUSIONS: Multiple active TrPs in the temporalis muscle were found, particularly in the anterior column and in the middle of the muscle belly. Bilateral posterior to anterior decreased distribution of PPTs in the temporalis muscle in women with CTTH was found. The locations of active TrPs in the temporalis muscle corresponded well to the muscle areas with lower PPT, supporting the relationship between multiple active muscle TrPs and topographical pressure sensitivity maps in the temporalis muscle in women with CTTH. PMID- 19542800 TI - No impaired hemoglobin oxygenation in forearm muscles of patients with chronic CRPS-1. AB - OBJECTIVES: Physiotherapy is considered an important treatment option in patients with upper limb complex regional pain syndrome type-1 (CRPS-1). In case of chronic CRPS-1, exercise therapy of the affected limb forms an important part of the physiotherapeutic program. We investigated whether muscle loading in chronic CRPS-1 patients is associated with impairments in muscle circulation of the forearm of the affected limb. METHODS: Thirty patients with chronic CRPS-1 unilaterally affecting their upper limbs, and 30 age-matched and sex-matched control participants were included in this study. Local muscle blood flow and hemoglobin oxygenation were measured by near infrared spectroscopy within the muscles of the forearm at rest, after 1-minute isometric handgrip exercises, and after arterial occlusion. Main outcome parameters were: local muscle blood flow, O2 consumption (mVO2), and postischemic reoxygenation (ReOx). RESULTS: We found no differences in baseline muscle blood flow, mVO2, and ReOx between the affected CRPS-1, unaffected CRPS-1, and control arms. After exercise, mVO2 of the affected CRPS-1 arms was not different from the clinically unaffected CRPS-1 arms. Furthermore, in comparison with the control arms, unaffected CRPS-1 arms showed no difference in mVO2 or ReOx. CONCLUSIONS: Muscle loading does not seems to be related to impairments in muscle oxygen uptake in forearm muscles of upper limbs affected by chronic CRPS-1. Our results suggest that exercise therapy can be safely used in physiotherapeutic training programs for chronic CRPS-1 of the upper limb. PMID- 19542801 TI - Fear avoidance and self-efficacy in relation to pain and sexual impairment in women with provoked vestibulodynia. AB - BACKGROUND: Provoked vestibulodynia is believed to be the most frequent cause of vulvodynia in women of childbearing age, with prevalence rates of up to 12% in the general population. Despite this high prevalence and the fact that vestibulodynia impacts negatively on quality of life, in particular sexual functioning, there has been a paucity of sound research to elucidate the condition's etiology. More specifically, few studies have focused on the role of psychologic factors in the experience of vulvo-vaginal pain and associated sexual impairment. OBJECTIVES: The present study aimed to determine the extent to which fear avoidance variables (catastrophizing, anxiety, fear of pain, hypervigilance) and self-efficacy differentially influenced changes in levels of induced and intercourse pain and also associated sexual dysfunction in these women. METHODS: Data were obtained from 75 vestibulodynia participants who completed a gynecologic examination, structured interview, and standardized questionnaires. RESULTS: The results of regression analyses revealed that higher catastrophizing, fear of pain, and hypervigilance in addition to lower self-efficacy together accounted for 15% of the variation in increased intercourse pain intensity. Among these, only catastrophizing contributed unique variance to intercourse pain. Results also showed that higher state anxiety and fear of pain (escape/avoidance) and also lower self-efficacy explained 22% of the variation in women's sexual impairment. However, only self-efficacy was found to be an independent correlate of sexual impairment. CONCLUSION: Findings support a theoretical model of vestibulodynia as a pain disorder influenced among others by cognitive and affective factors. PMID- 19542802 TI - Anticonvulsants for neuropathic pain: gaps in the evidence. AB - OBJECTIVE: To summarize the available evidence on the use of anticonvulsant drugs for the treatment of various conditions of neuropathic pain. METHODS: This is a systematic review on quantity and quality of evidence for using anticonvulsants in the management of neuropathic pain. Medical Subject Heading terms searched were "anticonvulsants" and "pain." Data sources used were the Cochrane Controlled Trials Register, Cochrane Database of Systematic Reviews, Medline 1966 to May 2006, Embase 1980 to May 2006, and CINAHL 1982 to May 2006. Additional studies were identified by hand searching the reference lists of retrieved papers and by autoalerts subsequent to May 2006. Randomized controlled trials, systematic reviews, and meta-analyses included. Non-English papers excluded. Evidence and evidence gaps with regard to specific conditions and drugs identified. RESULTS: Concise summary of all existing evidence or lack thereof was produced. A succinct table is presented of the efficacy of specific drugs for specific conditions and the nature and quality of evidence. The paper outlines the incidence of specific neuropathic pain conditions within various population groups and assesses the quantity of available evidence in regard to the frequency that those conditions are likely to occur. DISCUSSION: Gaps in the evidence are striking. Recommendations directly supported by the evidence by drug and by condition are made. PMID- 19542803 TI - Effectiveness of interventions of specific complaints of the arm, neck, or shoulder (CANS): musculoskeletal disorders of the hand. AB - OBJECTIVES: The aim of this study was to provide an evidence-based overview of the effectiveness of (conservative and surgical) interventions for the 4 specific pain disorders of the hand: trigger finger, primary Raynaud's phenomenon, Dupuytren disease, and De Quervain's disease. This information can help clinicians in the selection of interventions in daily practice, and may give direction to future research. METHODS: Relevant review publications and randomized clinical trials (RCTs) in PubMed were searched. Data extraction and quality assessment were performed. To summarize the results of the included reviews and RCTs, a best-evidence synthesis was used. RESULTS: For primary Raynaud's phenomenon (1 review, 20 RCTs), we found strong evidence for calcium channel blockers and moderate evidence for laser therapy. Limited evidence was found for Ketanserin, Prozasin, Buflomedil, transdermal glyceryl trinitrate patches, Ginkgo biloba, and behavioral treatment with temperature feedback. Other interventions did not show clear favorable treatment effects. For Trigger finger one very small RCT was found that showed limited evidence for steroid injection. For Dupuytren disease (4 RCTs) limited evidence was found in favor of use of staples versus sutures in the Dupuytren's surgery, and for intermittent compression on the postoperative hand after surgery. For other interventions no clear positive effects could be demonstrated. For De Quervain's disease (2 RCTs), we found no efficacy of Nimesulide as addition to a Triamcinolone injection, and no clear differences between a corticosteroid injection and a splint in pregnant patients or patients breast-feeding. DISCUSSION: Well-designed and well-conducted RCTs are clearly needed in this field. PMID- 19542806 TI - Ambulatory care management. Introduction. PMID- 19542807 TI - The impact of the electronic health record on an academic pediatric primary care center. AB - Many hospitals and practices are transitioning to electronic health records (EHR), but there is little information on the impact on patient care in a busy pediatric academic setting. The objective of this study was to determine the impacts of EHR on documentation, clinical processes, billing, ancillary staff responsibilities, scheduling, and cycle time. A descriptive study to assess the impact of EHR implementation and subsequent use on documentation, clinical processes, and patient access and flow was performed in a large urban academic pediatric primary care health center. Six months and 2 years after implementation, EHR impact on documentation was collected and compared with baseline value by measuring the percentage of charts with problem lists present. Several clinical improvement processes were collected at baseline and 6 months later including medication refill turnaround time, percentage of charts without attending signature at 3 days, and type of appointment billed on ill visits. The volume of appointments and cycle time were measured at regular intervals from baseline to 2 years after implementation. The percentage of paper charts attempted to be pulled for patient visits was obtained at baseline and 1 year later. Of the 500 charts audited before implementation, and 25 charts audited 6 months and 2 years after the implementation, the percentage of the presence of problem lists improved from 29% to 84%. Medication refill turnaround time improved from an average of 48 hours to 12 hours. Charts with incomplete documentation at 3 days postvisit decreased from 3% to 1.6%. Visit coding for detailed level visits (99214) increased by 13% and for problem-focused visits (99212) decreased by 7%, resulting in increased revenue collected. Medical records support staff needs decreased from 1 full-time equivalent to 0.5 full time equivalent. One year after the EHR implementation, the medical records staff pulled and refiled 5.2% of paper charts compared with 100% at baseline. Despite plans to return to full volume of scheduling patients by 4 weeks postimplementation, volume continued to be reduced by 10% for 3 additional months because of user inefficiency and high number of new learners/users. Patient cycle time was increased from 76 minutes preimplementation to 119 minutes immediately postimplementation and decreased to 85 to 90 minutes 2 years later. EHR can be successfully implemented in a large urban academic pediatric healthcare center. EHR implementation improved documentation of patient care, improved clinical processes, and resulted in increased revenue. However, the implementation of the EHR also led to short-term decreased appointment availability and a persistent longer cycle time. Ongoing information system training support is a key for maintaining efficiency due to the large number of new learners. PMID- 19542808 TI - Improvement in the family-centered medical home enhances outcomes for children and youth with special healthcare needs. AB - Family-centered, coordinated, comprehensive, and culturally competent care for children and youth with special healthcare needs is a national priority. Access to a primary care medical home is a US Maternal and Child Health Bureau performance measure. Most primary care practices lack methods by which to partner with families and improve care. Gaps remain in the number of children with access to a high-quality medical home. The Medical Home Index and Medical Home Family Index and Survey resulting from 10 pilot practices reveal improvements in practice capacity and subsequently in child and family outcomes. PMID- 19542809 TI - Care management for children with special needs: part I: the role of health plans. AB - This study documents screening methods and services provided by health plan case managers for high need children in a Washington State health plan. Enrollees were screened to identify 315 children who had or were at risk of developing a chronic condition and were high users of health services. From this group, 46 children/families could be contacted and needed case management. Services included assessment of physical/social needs, patient education, referral to community resources, and benefit utilization. These services were different from care coordination provided in primary care practices. PMID- 19542810 TI - Care management for children with special needs: Part II: the role of primary care. AB - This study documents care management services in 2 pediatric clinics for children with or at risk for a chronic condition during 8 months in 2005. Patients were identified by the clinic staff from a list provided by the health plan of patients at risk for or with a chronic condition. Care management services were documented for 161 of 189 selected patients. Services included family support, condition management, medical equipment management, and referrals to specialty care. Pediatric clinical care management activities directly relate to patient care and are complementary to, not duplicative of, case management provided by health plan managers. PMID- 19542811 TI - Ambulatory specialist use by nonhospitalized patients in us health plans: correlates and consequences. AB - Approximately 7 of 10 (and 95% of the elderly) people in US health plans see one or more specialists in a year. Controlling for extent of morbidity, discontinuity of primary care physician visits is associated with seeing more different specialists. Having a general internist as the primary care physician is associated with more different specialists seen. Controlling for differences in the degree of morbidity, receiving care from multiple specialists is associated with higher costs, more procedures, and more medications, independent of the number of visits and age of the patient. PMID- 19542812 TI - Healthcare intensity is associated with lower ratings of healthcare quality by younger adults. AB - Previous studies suggest a negative relationship between regional intensity of healthcare services and Medicare patients' perceptions of healthcare quality. However, no studies to date have examined the relationship across a broad age range of the population. We describe the responses of 64 088 respondents aged 19 to 69 years to an online interactive patient assessment and feedback system. We learn that in higher-intensive regions, specialty use is higher regardless of patient illness burden and that overall quality of care was lower in higher intensity regions. The findings that "more is not better" previously associated with Medicare patients seems to apply across a much broader age segment of the population. PMID- 19542813 TI - Differences in risk-adjusted mortality between medicaid-eligible patients enrolled in medicare advantage plans and those enrolled in the veterans health administration. AB - BACKGROUND: We compared risk-adjusted mortality rates between Medicaid-eligible patients in the Medicare Advantage plans ("MA dual enrollees") and Medicaid eligible patients in the Veterans Health Administration ("VHA dual enrollees"). METHODS: We used the Death Master File to ascertain the vital status of 1912 MA and 2361 VHA dual enrollees. We used Cox regression models to estimate hazard ratios (HRs) with 95% confidence intervals (CIs). RESULTS: The 3-year mortality rates of VHA and MA dual enrollees were 15.8% and 19.0%, respectively. The adjusted HR of mortality in the MA dual enrollees was significantly higher than in the VHA dual enrollees (HR, 1.260 [95% CI, 1.044-1.520]). This was also the case for elderly patients and those from racial/ethnic minority groups. CONCLUSIONS: The VHA had better health outcomes than did MA plans. The VHA's performance is reassuring, given its emphasis on equal access to healthcare in an environment that is less dependent on patient financial considerations. PMID- 19542814 TI - Developing a prospective payment system based on episodes of care. AB - A patient-centered approach to defining episodes of care around a hospitalization can provide the basis for creating expanded bundles of services that can be used as the basis of payment. Paying by episodes of care strengthens the incentive to providers to deliver care efficiently. A hospital-based episode of care prospective payment system can be phased in over time by gradually expanding the services and the time period included in the episode. Establishing equitable prospective episode payment amounts requires that the severity of illness of the patient during the hospitalization and the chronic disease burden of the patient be taken into account. PMID- 19542816 TI - TROT line: live and direct from the Republic of Texas. PMID- 19542815 TI - The 2008 Erice statement toward a more humanistic oncology. PMID- 19542817 TI - What if ...: you need to help someone who's a lawyer? PMID- 19542818 TI - Why don't gastroenterologists follow colon polyp surveillance guidelines?: results of a national survey. AB - GOALS: To measure knowledge and acceptance of colon polyp surveillance guidelines among gastroenterologists. BACKGROUND: Gastroenterologists often perform surveillance colonoscopy sooner than recommended by guidelines. Lack of knowledge may be an important factor, but gastroenterologists could also simply disagree with guideline recommendations. STUDY: A 24-item multiple-choice survey was developed from the 2003 multisociety colorectal cancer screening and surveillance guideline. The survey was distributed to practicing gastroenterologists preparing for the gastroenterology board recertification examination at 2 major national gastroenterology board review courses. For each question, subjects were given a clinical scenario and asked: (1) the guideline recommendation for the scenario; (2) their usual practice in the scenario; and, (3) if they definitely knew the recommendation or were simply guessing at the correct answer. If a respondent knew the recommendation but differed in their usual practice, this was considered disagreement with the recommendation. RESULTS: The survey was completed by 57.1% (116/203) of gastroenterologists preparing for 2004 recertification. Seventy eight percent reported that they were familiar with the 2003 guideline, though only 57% reported that guidelines were "very influential" in their practice. Many did not correctly answer questions on the recommended interval for hyperplastic polyps (12%), 2 small adenomas (36%), 3 small adenomas (49%), and adenoma with high-grade dysplasia (41%). Of gastroenterologists who knew the guideline recommendations, up to 76% disagreed with the recommendations and chose to perform surveillance sooner than recommended. CONCLUSIONS: Though many gastroenterologists lack knowledge about guideline recommendations for colon polyp surveillance, even those who know the recommendations often ignore them and perform surveillance colonoscopy sooner than recommended. PMID- 19542819 TI - Journal of the American Academy of Child & Adolescent Psychiatry. In this issue. PMID- 19542820 TI - Youth binge drinking: progress made and remaining challenges. PMID- 19542821 TI - Medicalize depression, not sadness. PMID- 19542822 TI - Should screening for depression among children and adolescents be demedicalized? PMID- 19542823 TI - Conceptualizing autism: the role for emergence. PMID- 19542824 TI - Cortisol levels in childhood and psychosis risk in late adolescence. PMID- 19542825 TI - Long-term durability of cognitive behavioral therapy gains for pediatric obsessive-compulsive disorder. PMID- 19542827 TI - Pediatric Research--the evolution continues. PMID- 19542828 TI - Mining the x-chromosome for disease genes by deep resequencing. PMID- 19542829 TI - Endo-laparoscopic reduction and resection of gastroduodenal intussuception of gastrointestinal stromal tumor (GIST): a synchronous endoscopic and laparoscopic treatment. AB - Gastrointestinal stromal tumor (GIST) commonly occurs in the stomach. We would like to report an uncommon presentation of gastric GIST with gastroduodenal intussuception. A patient with known history of gastric GIST at fundus for 10 years presented to the casualty department with recurrent epigastric pain, deranged liver function, and hyperamylasemia. Computed tomography of the abdomen showed intussuception of the gastric GIST into duodenum. Emergency operation with synchronous endoscopic reduction and laparoscopic wedge resection was performed and patient had uneventful recovery. This simultaneous endoscopic and laparoscopic treatment should be considered for this rare complication of GIST. PMID- 19542830 TI - Splenic rupture after colonoscopy: a case report. AB - Iatrogenic splenic tear after a colonoscopy is a rare complication. Conditions predisposing to splenocolic adhesions such as previous abdominal surgery, pancreatitis, or inflammatory bowel disease may increase the risk of this injury after colonoscopy. We present a case of a 47-year-old woman with an iatrogenic splenic tear after a routine colonoscopy, who had several of these predisposing factors and required an urgent splenectomy. PMID- 19542831 TI - Abdominal aortic pseudoaneurysm managed with endovascular stent graft. AB - Angioplasty, stenting, endovascular stent grafts, and other minimally interventional techniques are becoming common techniques used for a myriad of vascular pathology. As the technology, comfort level, and technical expertise improve, the envelope of overuse is being approached or possibly superceded. We present an unusual complication of pancreatitis, pseudoaneurysm of the abdominal aorta, which was successfully treated with an endovascular stent graft. PMID- 19542832 TI - Transrectal intraperitoneal assess with transanal endoscopic operation (TEO) device in pig model. AB - BACKGROUND: Gaining peritoneal access with subsequent safe closure is a prerequisite for natural orifice translumenal surgery (NOTES). We explored the possibility of transanal endoscopic operation (TEO) device to perform transrectal peritoneoscopy in a pig model. OBJECTIVE: Performing transrectal peritoneoscopy with TEO device in pig model. METHODS: Two pigs were used for transrectal peritoneoscopy. A 40-mm rectoscope was inserted via the anus after the induction of general anesthesia. Proctotomy was then created with diathermy and the endoscope was passed through the pararectal space into the peritoneal cavity. Proctotomy was closed with absorbable suture after completion of peritoneoscopy. RESULTS: Completion of the procedures was achieved in 2 pigs. Both pigs survived for more than 30 days. Necropsy revealed completely healed rectum with no evidence of leakage or abscess formation. Adhesions around the colostomy site were minimal. CONCLUSIONS: In conclusion, incorporation of TEO system is safe and useful in animal model for creation and closure of proctotomy for natural orifice translumenal surgery in transrectal access, further experiment should be performed to validate the possibility of application in human. Potential complications need to be addressed and well documented. PMID- 19542833 TI - Endoscopic thyroidectomy with the da Vinci robot system using the bilateral axillary breast approach (BABA) technique: our initial experience. AB - INTRODUCTION: Robotic surgery is useful in areas with difficult access like the pelvis. The ideal indications for robotic surgery are still to be established. The neck area, especially the thyroid gland poses a difficult challenge for many endoscopic surgeons. Robotic surgery is useful in this area due to its excellent magnification and endowrist function. We present our initial experience with robotic endoscopic thyroidectomy using the bilateral axillary breast approach (BABA). METHODS AND RESULTS: Between March and May 2008, 15 patients diagnosed with papillary thyroid cancer underwent robotic-assisted endoscopic thyroidectomy using the BABA technique. The mean operating time was 218 minutes. There was a steady decrease in operative time from the initial case to the 15th case. The blood loss was minimal. The recurrent laryngeal nerve and parathyroid glands were identified in great detail with ease and preserved in all cases. There were no postoperative complications in any case. CONCLUSIONS: Robotic endoscopic thyroidectomy using the BABA technique is a feasible procedure and can be performed safely. It provides an excellent operative field view enabling easy identification of vital structures. It also gives the desired cosmetic results and minimal postoperative pain similar to conventional endoscopic thyroid surgery using the BABA technique. PMID- 19542834 TI - Laparoscopic-assisted stomach-partitioning gastrojejunostomy for the palliation of gastric outlet obstruction from antral gastric cancer. AB - Conventional gastrojejunostomy is performed for gastric outlet obstruction; however, we have experience of some patients for whom oral intake was not adequately restored. Open stomach-partitioning gastrojejunostomy is a useful technique for the relief of unresectable and obstructing antral gastric cancers. We herein report the successful laparoscopic application of this technique in 2 patients: a 62-year-old male and a 65-year-old female with obstructing antral gastric cancer. There were no operative complications, and blood loss was minimal. Oral fluid intake was resumed 3 and 4 days postoperatively. Chemotherapy by oral administration was possible for each patient. There was no recurrence of the gastric outlet obstruction in either patient up until they died 6 and 10 months after surgery. Laparoscopic stomach-partitioning gastrojejunostomy is a safe and useful technique for treating unresectable and obstructing antral gastric cancer, allowing the possibility of adequate oral intake and permitting the administration of anticancer drugs. PMID- 19542835 TI - Laparoscopic intestinal derotation: original technique. AB - The intestinal derotation technique, introduced by Cattel and Valdoni 40 years ago, is carried out using a laparoscopic procedure, which is described here for the first time. The method is effective in the treatment of malign lesions of the III and IV duodenum and during laparoscopic subtotal colectomy with anastomosis between the ascending colon and the rectum. Ultimately, the procedure allows for the verticalization of the duodenal C and the anterior positioning of the mesenteric vessels, facilitating biopsy and resection of the III and IV duodenal portions and allowing anastomosis of the ascending rectum, avoiding both subtotal colectomy and the risk of torsion of the right colic loop. Although the procedure calls for extensive experience with advanced video-laparoscopic surgery, it is both feasible and repeatable. In our experience we have observed no mortality or morbidity. PMID- 19542836 TI - Endoscopic subcutaneous mastectomy for the treatment of gynecomastia: a report of 65 cases. AB - The aim of this study is to introduce the endoscopic subcutaneous mastectomy without skin excision as the standard surgical technique for grade IIB and III gynecomastia. Endoscopic subcutaneous mastectomy was performed successfully in 125 breasts of 65 patients with Simon's grade IIB and III gynecomastia. The volume of gland resected in the 125 breasts was 80 to 300 mL, with the mean of 146 mL. The operation time was 65 to 155 minutes, with the mean of 82 minutes. There were a few operative complications, including partial nipple necrosis in 2 cases and subcutaneous hydrops in 1 case; but no complete nipple necrosis, subcutaneous emphysema, postoperative active bleeding, local skin necrosis, or operation-side infection occurred. Satisfactory chest contour was gained in all cases without any abnormality, skin redundancy, or recurrence during the follow up of 3 to 36 months. Endoscopic subcutaneous mastectomy is distinctive and practicable in manipulation as well as safe and can get esthetic effect. It is a new choice for the treatment of gynecomastia. PMID- 19542837 TI - Fibrin sealant (Tissucol) for the fixation of hiatal mesh in the repair of giant paraesophageal hernia: a case report. AB - INTRODUCTION: The use of hiatal meshes for the repair of giant paraesophageal hernias (GPH) is associated with a significantly decreased rate of recurrences compared with mesh free techniques. Many surgeons refrain from mesh implantation at the gastroesophageal junction owing to reported complications, such as mesh migration, strictures, and risks of tack or suture placement. This case report presents the laparoscopic application of a titanium-coated mesh (TiSure, GfE, Germany) designated for hiatal repair, with fibrin sealant fixation (Tissucol, Baxter, Austria) in a patient with GPH. METHODS: A patient (male, 59 y) presented at our outpatient department with a 3-year history of epigastric pain and decreasing lung capacity. A GPH with an intrathoracic upside-down stomach had already been radiologically diagnosed 3 years before admission. In elective laparoscopy, the stomach was repositioned and the crura of the diaphragm were approximated with nonresorbable sutures. The defect was reinforced with a preshaped titanium-coated mesh and fibrin sealant (2 mL) applied with a 45 degree angled tip laparoscopic spraying device. No perforating fixation device was used for mesh fixation itself. The patient was discharged on the seventeenth postoperative (postOP) day. The clinical follow-up included the assessment of postOP pain with a visual analog score and a confirmative computed tomography scan 6 months after surgery. RESULTS: The patient has fully recovered, showing no recurrence or adverse effects 1 year postOP. DISCUSSION: Based on previous good results from own experimental trials, the mesh sealing approach in hiatal hernia repair was performed clinically, yielding an excellent result in this case. Multicenter trials to assess the full impact of FS mesh fixation in combination with macroporous hiatal meshes seem mandatory. PMID- 19542838 TI - Laparoscopic resection for a gastric pheochromocytoma: a rare presentation of an uncommon tumor. AB - Pheochromocytoma is a rare tumor of the stomach, having only been reported 6 times in the published literature. We report the first case of gastric pheochromocytoma treated by laparoscopic partial gastrectomy. PMID- 19542839 TI - Direct percutaneous endoscopic duodenostomy: a case report. AB - A 78-year-old male, who had undergone left upper lobectomy because of traumatic pulmonary contusion at the age of 25 years, succumbed to left hemiplegia with impaired consciousness that was caused by right putaminal hemorrhage. Aspiration pneumonia was complicated and he was set on tracheostomy and tube feeding through a nasogastric tube. Although it was apparent that enteral nutrition through the percutaneous endoscopic gastrostomy was appropriate for him, performing a percutaneous endoscopic gastrostomy seemed impossible as the computed tomography revealed that the elevated stomach was located behind the heart. In this patient, we successfully performed a direct percutaneous endoscopic duodenostomy without any complication. PMID- 19542843 TI - Laparoscopic splenectomy: 16 years since Delaitre with review of current literature. AB - Since Delaitre reported his first laparoscopic splenectomy it has been accepted universally and today it is probably regarded as a Gold standard in removal of benign and normal sized spleens. We discuss the indications, advantages, techniques, and recent advances of Laparoscopic splenectomy after 16 years of Delaitre. PMID- 19542840 TI - Clinical prospect of applying the chemical bile duct embolization to achieve a chemical hepatectomy in the treatment of highly selected hepatolithiasis. AB - The high recurrence rate of hepatolithiasis, together with the high operative risk of hepatectomy for specifically located stones shows that an effective treatment for intrahepatic stones has not been settled upon. It is commonly accepted that a diseased biliary duct mucosa is a prerequisite for the development of intrahepatic stones, and that segmental biliary obstruction is able to induce hepatic atrophy, fibrosis, and "self-cut" the obstructed hepatic segment. Therefore, we previously put forward the hypothesis that performing deliberate chemical bile duct embolization (CBDE) to induce a segmental chemical hepatectomy might be the way of treating hepatolithiasis. In this study, we review the relative experimental basis for CBDE, preliminary report on its clinical use in 2 patients, and speculate on its future application. To completely embolize a diseased biliary duct, absolute ethanol or phenol is firstly used to ablate the biliary mucosa and eradicate biliary bacteria. Subsequently, cyanoacrylate or tissue adhesive glue is used to permanently fill the duct lumen, occupying the space where the stones would have formed. Our prior laboratory investigations and preliminary clinical treatments have confirmed that this combination of embolization agents could not only achieve the desired aim of preventing stone recurrence but could also lead to complete atrophy of the targeted hepatic segment, thereby achieving a chemical hepatectomy. In the future, CBDE is likely to help in resolving the problem of calculous recurrence and thereby reduce the incidence of surgical reintervention and endoscopic stone extractions, which are so frequently needed in patients with hepatolithiasis. Also chemical hepatectomy might provide a new less-invasive hepatectomy method, especially for the more difficult resections of the caudate or right posterior lobes. PMID- 19542844 TI - Prevention and treatment of abdominal wall bleeding complications at trocar sites: review of the literature. AB - Abdominal wall bleeding may complicate any laparoscopic procedure. Piercing or laceration of vessels transversing the abdominal wall during trocar placement is generally the cause. Bleeding may occur at the very beginning of the surgery but, in some cases, it may go unrecognized for a while complicating the operation and the postoperative course. Planned and careful trocar placement can prevent most of these instances that otherwise can be readily managed avoiding severe morbidity. PMID- 19542845 TI - Results of laparoscopic Heller myotomy for extreme megaesophagus: an alternative to esophagectomy. AB - Heller myotomy is recognized as the optimal treatment for achalasia. However, treatment of the markedly dilated esophagus has been debated in the literature. Although esophagectomy has been the standard treatment historically, several studies have examined successful treatment of achalasia with laparoscopic Heller myotomy in the setting of a markedly dilated esophagus (>6 cm). Patients with extreme megaesophagus (>10 cm) are often treated with esophagectomy. We report the successful treatment of 4 patients with extreme megaesophagus with laparoscopic Heller myotomy. Three of the 4 patients also had Toupet fundoplication. The average esophageal diameter was 11.2 cm (10 to 12 cm). In addition to severe dysphagia, all patients had preoperative signs, symptoms, and radiographic evidence of esophageal compression of their heart and lungs. All patients reported relief of their preoperative symptoms. Esophagectomy has not been required to maintain adequate clinical results in any of our patients. We conclude that laparoscopic Heller myotomy is an appropriate alternative to esophagectomy and can be offered to patients with extreme megaesophagus. PMID- 19542846 TI - Comparison of laparoscopic and open gastrectomy on cancer cells exfoliating from the cancer-invaded serosa. AB - BACKGROUND: Whether laparoscopic gastrectomy may reduce the frequency of gastric cancer cells exfoliating from the cancer-invaded serosa remains unclear. This study aimed to compare the detection of free gastric cancer cells in the peritoneal cavity during laparoscopic and open gastrectomy. METHODS: Intraoperative peritoneal washings were collected from 63 gastric cancer patients undergoing laparoscopic gastrectomy and 61 patients undergoing open surgery. Hematoxylin and eosin staining and real time reverse transcription-polymerase chain reaction were used to examine the free cancer cells. RESULTS: The postoperative positive rates of free cancer cells detected by cytologic and real time reverse transcription-polymerase chain reaction were 39.68% and 44.26% in the laparoscopic and open groups, respectively. The depth of tumor invasion, area of invaded serosa, regional lymph node involvement, and higher tumor node metastasis staging were significantly associated with the presence of free cancer cells. CONCLUSIONS: The laparoscopic techniques used in gastric cancer surgery were not associated with a greater risk for the intraperitoneal dissemination of cancer cells than conventional techniques. PMID- 19542847 TI - Radiofrequency ablation for metachronous hepatic metastases from gastric cancer. AB - The objective of this study was to evaluate the results of radiofrequency ablation (RFA) for the treatment of metachronous hepatic metastases from gastric adenocarcinoma. Between January 2000 and February 2008, we retrospectively reviewed 7 cases for which RFA was performed for treating metachronous hepatic metastases after resection of the primary gastric adenocarcinoma. The median period between curative gastrectomy and metachronous hepatic metastasis was 14 months (range: 6 to 48 mo). The median survival of the patients was 11.0 months (range: 5.5 to 39.2) after the first detection of hepatic metastases and 10.0 months (range: 5 to 38.2) after the first RFA. Hepatic metastases recurred 3 to 21 months after RFA in all patients. A second RFA was performed for a single recurrent hepatic metastasis in 1 patient and this patient survived more than 3 years without recurrence until the time of this study. In conclusion, the efficacy of RFA alone for metachronous hepatic metastases from gastric adenocarcinoma was disappointing due to multiple intrahepatic recurrences. Combination therapy such as systemic chemotherapy or hepatic arterial infusion chemotherapy adjuvant to RFA would more reasonable for treating hepatic metastases from gastric cancer. PMID- 19542848 TI - Monopolar electrosurgery on the extrahepatic bile ducts during laparoscopic cholecystectomy: an experimental controlled trial. AB - INTRODUCTION: The purpose of this study is to investigate the occurrence of thermal injury in the extrahepatic bile ducts when monopolar electrosurgery is used to perform dieresis of the cystic duct and cystic vessels in laparoscopic cholecystectomy. METHODS: Female pigs (n=40) of the Large White breed were separated into 2 groups of 20 animals. In the experimental groups, dieresis of the cystic duct and cystic vessels was performed with monopolar electrosurgery using a hook-like dissector using a power setting of 20 W, whereas in the control group this procedure was performed with a pair of Metzenbaum scissors disconnected from any kind of thermal energy source. Occurrence of distal thermal injury was evaluated on 2 occasions, on the 3rd and 28th days postoperatively. It consisted of exploratory laparotomy, cholangiography and both macroscopic and microscopic examination of the surgical specimen, which included cystic duct stump, hepatic duct and choledochus. RESULTS: The presence of distal thermal injury, classified as second degree, 1.2 mm in the portion near the clipping area, was observed in only one of the cystic duct stumps after microscopic examination, without statistical significance. No thermal injury was observed in the extrahepatic bile ducts. CONCLUSIONS: Monopolar electrosurgery produced negligible thermal injury in the extrahepatic bile ducts after laparoscopic cholecystectomy. PMID- 19542849 TI - Migration of plastic biliary stents and endoscopic retrieval: an experience of three referral centers. AB - BACKGROUND: Proximal or distal migration of a plastic biliary stent is uncommon, but its management can be a technical challenge to the pancreatobiliary endoscopist. PATIENTS AND METHODS: All cases (n=51) of proximally and distally migrated plastic biliary stents over an 8-year period at 3 referral pancreaticobiliary centers were included in this retrospective study. Indications for stenting, risk factors for migration, presentation of migration, and various techniques used for stent's retrieval are herein analyzed. RESULTS: Twenty-one proximal and 30 distal bile duct-migrated stents were identified. All patients with proximally and 17 (56.7%) with distally migrated stents were symptomatic. Choledocholithiasis, dilated common bile duct, short and large size stent were the main risk factors. The retrieval of proximally migrated stents was successful in 15 patients (71.4%) and in all symptomatic patients with distal migration. The retrieval techniques included forceps, Dormia basket, snare, Soehendra stent retriever, and balloon. One patient died of sepsis due to peritonitis from duodenal perforation from a distally migrated stent. CONCLUSIONS: Retrieval of a proximally migrated stent requires experience with different endoscopic devices. Moreover, distal migration needs attention because it can cause severe complications. PMID- 19542850 TI - The treatment of gallbladder dyskinesia based upon symptoms: results of a 2-year, prospective, nonrandomized, concurrent cohort study. AB - BACKGROUND: Gallbladder dyskinesia (GD) is controversial. We sought to determine the success rate of cholecystectomy or observation in treating patients with GD with intervention decisions based upon clearly defined symptoms. METHODS: Ninety three consecutive patients with documented GD were enrolled into a 2-year prospective study. Based upon the presenting symptoms categorized as either classic for gallbladder pathology or atypical, patients underwent cholecystectomy (classic) or observation (atypical). We defined dyskinesia as a cholecystokinin (CCK)-stimulated ejection fraction (EF) <35% on nuclear cholescintigraphy and a negative gallbladder ultrasound. RESULTS: Classic gallbladder symptoms were identified in 61 patients and an atypical presentation occurred in 32 patients. The EF with CCK stimulation was not significantly different between the groups (19+/-9% vs. 16+/-7%, P=0.12). Of those with atypical symptoms, 28% (9 out of 32) had resolution of their symptoms without surgery. About 72% (23 out of 32) had worsening or progressive symptoms that did not resolve during observation, and later underwent surgery. Of these, 57% (13 out of 23) had resolution of their symptoms after surgery, but 43% (10 out of 23) had no improvement. Of those with classic symptoms, 60 patients underwent laparoscopic cholecystectomy with resolution of symptoms in 58 (97%). Patients with classic symptoms were 22 times more likely to have relief after cholecystectomy (odds ratio 22.3, P=0.0002). Eight patients had their symptoms recur more than 1 year after surgery (3 atypical and 5 classic) such that at long-term follow-up, cholecystectomy had helped only 43% of the atypical patients and 88% of the classic patients. CONCLUSIONS: Classic biliary symptoms are more predictive of success after cholecystectomy in patients with GD than is EF. The symptoms that are most predictive of success after surgery are right upper quadrant pain, pain after meals, and reproduction of the pain after CCK administration. Patients with atypical symptoms are much less likely to have improvement after surgery and should be observed; however, recurrent or progressive symptoms should prompt intervention if all additional testing is negative. PMID- 19542851 TI - Laparoscopic cholecystectomy in older patients: clinical experience from 56 consecutive patients in a rural community hospital in Taiwan. AB - PURPOSE: Laparoscopic cholecystectomy (LC) is a safe and effective treatment for gallbladder disease in elderly patients. However, there is little convincing prospective data regarding the outcome of LC in patients above 75 years of age. METHODS: Fifty-six cholecystitis patients were enrolled. Group 1 consisted of 17 patients above 76-years-old, group 2 of 39 patients aged between 65 and 75 years. Intraoperative and postoperative parameters were measured. RESULTS: Hypertension, diabetes mellitus, and coronary artery disease were the most common medical comorbidities. Operation time, length of hospital stay, and incidence of complications did not differ significantly between the 2 groups. Blood loss did increase significantly in group 1. Similar results were noted in patients with acute cholecystitis. No mortality occurred. CONCLUSIONS: LC is a safe and effective treatment for older patients suffering from cholelithiasis, but these patients are at greater risk of blood loss. Preoperative coagulation profiles should be arranged. PMID- 19542852 TI - Surgical treatment for xanthogranulomatous cholecystitis: a report of 74 cases. AB - AIMS: To be more aware of the presence of xanthogranulomatous cholecystitis (XGC) and find a better surgical measure of its treatment. METHODS: Data from 74 cases of XGC treated between May 1996 and May 2008 at our hospital were retrospectively analyzed and reported here. Laparoscopic and laparotomy group were compared with respect to operative time, postoperative hospital stay, postoperative complication, etc. RESULTS: In the 74 cases, 47 underwent laparoscopic surgery, the rest 27 underwent laparotomy surgery. The mean operative time of laparotomy and laparoscopic cases were 113.9 minutes and 69.4 minutes, respectively, which shows statistically significant difference between the 2 groups (P<0.01). The postoperative hospital stay of the laparotomy and laparoscopic group is 18.3 days and 8.66 days, respectively (P<0.01). The converting rate of the laparoscopic group is 10.6%. CONCLUSIONS: Surgical treatment remains the most effective and feasible option for XGC. PMID- 19542853 TI - A prospective randomized, controlled study comparing low pressure versus high pressure pneumoperitoneum during laparoscopic cholecystectomy. AB - BACKGROUND: The increase in intra-abdominal pressure by insufflation of carbon dioxide during laparoscopy brings certain changes in function of organ systems and also leads to postoperative pain. Degree of intra-abdominal pressure is directly related with such change. Laparoscopic cholecystectomy can be performed at low pressure pneumoperitoneum. However, available space for dissection is less than the high pressure pneumoperitoneum. METHODS: Twenty-six patients for elective laparoscopic cholecystectomy were studied in a prospective, randomized, patient, and surgeon blinded manner. The intra-abdominal pressure was kept either in low pressure (8 mm Hg) or in high pressure (12 mm Hg). All patients underwent two dimensional echocardiography, pulmonary function test and color Doppler examination of lower limb vessels preoperatively and postoperatively. Arterial blood gas analysis and End Tidal CO2 monitored before insufflation, during surgery and after deflation. Pain score was measured by visual analog scale and surgeon's comfort level was recorded. Postoperative analgesia requirement, complications, and hospital stay were recorded. Student t test used for the statistical analysis. RESULTS: Both groups match for the demographic parameters. Four patients required conversion to high pressure. Intraoperative pO2 level, postoperative pain, analgesic requirement, pulmonary function, and hospital stay were favoring low pressure pneumoperitoneum in a statistically significant manner. There was no difference between 2 groups for duration of surgery, intraoperative, and postoperative complications. However, the technical difficulties were graded more (statistically nonsignificant) with low pressure pneumoperitoneum. CONCLUSIONS: An uncomplicated gall stone disease can be treated by low pressure laparoscopic cholecystectomy with reasonable safety by an experienced surgeon. Though surgeons experience more difficulty in dissection during low pressure pneumoperitoneum, it is significantly advantageous in terms of postoperative pain, use of analgesics, preservation of pulmonary function, and hospital stay. PMID- 19542854 TI - Second-look laparoscopy after laparoscopic relief of strangulated small bowel obstruction. AB - Laparoscopy is increasingly used for the treatment of complex small bowel obstructions (SBO). Conventional treatment of strangulated bowel is segmental resection. We have used second-look laparoscopy to preserve bowel in 4 cases of strangulated SBO. Of the 17 patients with bowel obstruction treated laparoscopically 4 had ischemic bowel. The obstructions were relieved and second look laparoscopy was performed 24 hours later. In 3 cases the bowel had largely regained its normal appearance; 1 case required resection based on persistent ischemia. An average of 20 cm of bowel was preserved per patient, and there were no complications in these 4 patients. In sum, we have shown second-look laparoscopy for strangulated SBO to be feasible, safe, and, in most cases, bowel conserving. PMID- 19542855 TI - Risk factors of postoperative infections in adults with complicated appendicitis. AB - BACKGROUND: Acute appendicitis is the most common surgical emergency. Infectious complications are uncommon but carry significant morbidity especially in complicated appendicitis. This study aimed to determine predictive factors of postoperative infections in adults with complicated appendicitis. METHOD: Retrospective review of clinical records from CDARS of Hong Kong Hospital Authority was performed in all patients aged 18 or above, operated with the diagnosis of gangrenous appendicitis, acute appendicitis with generalized peritonitis or peritoneal abscess from January 2003 to December 2005 in a regional hospital. Primary end point was postoperative infections, including wound infection and postoperative intra-abdominal abscess (PIAA). Independent predictors were determined by logistic regression analysis. RESULTS: One seventy three patients were operated with the diagnosis of complicated appendicitis. Postoperative infections occurred in 40 patients (23.1%). Thirty-two patients had wound infection and 10 patients had PIAA. Two patients had both infectious complications. There were 2 deaths. Diabetes mellitus was the only significant factor associated with PIAA in univariate analysis (odds ratio, 6.6; 95% confidence interval, 1.67-25.9). No independent factors can be identified in multivariate analysis. Successful laparoscopic appendicectomy was the only independent negative risk factor of wound infection in logistic regression. CONCLUSIONS: Laparoscopic appendicectomy is the first choice of procedure in complicated appendicitis. Patients with diabetes mellitus need close monitoring of postoperative infections. PMID- 19542856 TI - Evolution of laparoscopic colorectal surgery in Brazil: results of 4744 patients from the national registry. AB - BACKGROUND: Since its introduction, laparoscopic colorectal surgery has raised intense debate and controversies regarding its safety and effectiveness. METHODS: This multicentric registry reports the experience of 28 Brazilian surgical teams specializing in laparoscopic colorectal surgery. RESULTS: Between 1992 and 2007, 4744 patients (1994 men--42% and 2750 women--58%) were operated upon, with ages ranging from 13 to 94 years (average 57.5 y). Benign diseases were diagnosed in 2356 patients (49.6%). Most diseases were located in 50.7% of the left and sigmoid colon, 28.2% in the rectum and anal canal, 8.0% in the right colon, and diffuse 7.0%. There were 181 (3.8%) intraoperative complications (from 0% to 14%). There were 261 (5.5%) reported conversions to laparotomy (from 0% to 16.5%), mainly during the early experience (n=119 -59.8%). Postoperative complications were registered in 683 (14.5%) patients (from 5.0% to 50%). Mortality occurred in 43 patients (0.8%). Surgeons who performed less than 50 cases reported similar rates of intraoperative (4.2% vs. 3.8%; P=0.7), postoperative complications (20.8% vs. 14.3%; P=0.07), and mortality (1.0% vs. 0.9%; P=0.5), but the conversion rate was higher (10.4% vs. 5.4%; P=0.04). Two thousand three hundred and eighty-nine (50.4%) malignant tumors were operated upon, and histologic classification showed 2347 (98%) adenocarcinomas, 30 (0.6%) spinocelular carcinomas, and 12 (0.2%) other histologic types. Tumor recurrence rate was 16.3% among patients followed more than 1 year. After an average follow up of 52 months, 19 (0.8%) parietal recurrences were reported, 18 of which were in port sites and 1 in a patient with disseminated disease. There was no incisional recurrence in the ports used to withdraw the pathologic specimen. Compared with other registries, there was a 75% increase in the number of groups performing laparoscopic colorectal surgery and a decrease in conversions (from 10.5% to 5.5%) and mortality (from 1.5% to 0.9%) rates. CONCLUSIONS: (1) The number of patients operated upon increased expressively during the last years; (2) operative indications for benign and malignant diseases were similar, and diverticular disease of the colon comprised 40% of the benign ones; (3) conversion and mortality rates decreased over time; (4) surgeon's experience did not influence the complication rates, but was associated with a lower conversion; and (5) oncologic outcome expressed by recurrence rates showed results similar to those reported in conventional surgery. PMID- 19542857 TI - Laparoscopic colon resection of benign polyps: high grade dysplasia on endoscopic biopsy and polyp location predict risk of cancer. AB - BACKGROUND: To ensure that an oncologic resection is not compromised, laparoscopic surgery is not recommended for colon cancer patients during the learning curve. Although patients with colon polyps are frequently selected as "learning cases," several studies have found a high incidence of occult cancers in this subset of patients questioning the safety of this approach. A retrospective review was performed of laparoscopic resections for colonic polyps to determine the incidence of occult cancer and, clinical and pathologic factors predictive of cancer. METHODS AND RESULTS: From January 2004 and September 2007, 44 colectomies were performed for colonic polyps. Data from a prospective database was examined and supplemented by pathology reports. Of 44 patients, operative conversion was 5% with a 4-day median length of stay, a morbidity of 20% with no mortalities. All patients had clear surgical margins with a median of 12 lymph nodes detected. An occult cancer was detected in 6 of 44 cases (14%). High-grade dysplasia on endoscopic biopsy and polyp location distal to the splenic flexure was associated with a significantly (P<0.05) increased risk of cancer. Forty-three percent of cancers were detected in polyps with high-grade dysplasia compared with 8% of polyps without dysplasia on biopsy. Polyps distal to the splenic flexure had a 43% rate of occult cancers detected compared with 8% in polyps proximal to the splenic flexure. CONCLUSIONS: Colonic polyps with high grade dysplasia on endoscopic biopsy and polyps located distal to the splenic flexure are associated with the highest risk of cancer and may not be appropriate cases during the learning curve. PMID- 19542858 TI - Laparoscopic versus open splenectomy for hypersplenism secondary to liver cirrhosis. AB - BACKGROUND: Since the first laparoscopic splenectomy (LS) in 1991, LS has become the gold standard for the removal of normal-to-moderately enlarged spleens in benign conditions. Compared with open splenectomy (OS), fewer postsurgical complications and better postoperative recovery were observed, but it is contraindicated for hypersplenism secondary to liver cirrhosis owing to technical difficulties associated with splenomegaly, well-developed collateral circulation, and increased risk of bleeding. With the improvements of laparoscopic technique, the concept is changing. METHODS: OS and LS performed for hypersplenism secondary to liver cirrhosis at our institution were analyzed. Relationships between postoperative increases in platelet counts, white blood cell counts, hemoglobin, and liver function were examined. Perioperative data of LSs were compared with those of OSs, including operative time, blood loss, excised spleen weight, complications, and hospital stays. RESULTS: A total of 216 splenectomies (135 OS and 81 LS) were performed from April 1999 to March 2007. Five laparoscopic cases were converted to open surgery owing to operative bleeding or bleeding of splenic fossa. The other 76 patients were performed LSs successfully. No major operative complications occurred. There was no operative death. Excised spleen weight >400 g was present in 56% of cases in this series. At 7 days postoperatively, the platelet counts, white blood cell counts, and hemoglobin significantly increased after open and laparoscopic surgeries, and increase of alanine aminotransferase, aspartate aminotransferase, total bilirubin, and directed bilirubin of LS were significantly different with open cases. Operation times of LS and OS were 2.9+/ 0.7 hours and 2.6+/-0.6 hours, respectively. Blood losses were 150.6+/-135.4 mL and 633.8+/-340.3 mL (P<0.01), excised spleen weights were 585.7+/-184.6 g and 591.1+/-153.4 g (P>0.05), and hospital stay were 8.2+/-2.0 days and 11.9+/-3.8 days (P<0.01). Operative associated complications were noted in both LS and OS. Less blood loss, shorter hospital stay, and less impairment of liver function were observed in LS than OS. CONCLUSIONS: LS is feasible, effective, and safe procedures for hypersplenism secondary to liver cirrhosis and contributes to less impairment of liver function, less blood loss, and shorter hospital stay. PMID- 19542860 TI - Inguinal hernia repair in a community hospital setting--have attitudes changed because of laparoscopy? A review of a general surgeon's experience over the last 5 years. AB - OBJECTIVES: The laparoscopic transabdominal preperitoneal (TAPP) approach for inguinal hernia repair is well documented in numerous studies as an excellent choice when performed by an experienced surgeon. In this report we wish to evaluate our experience of TAPP laparoscopic inguinal repair performed in a Community Hospital over the last 5 years, focusing on the feasibility of the technique and the incidence of complications when performed by general surgeons in this setting. We also wish to report and discuss how our attitudes concerning inguinal hernia repair have changed since we adopted the laparoscopic approach. MATERIALS AND METHODS: Between January 2003 and January 2008 a total of 193 patients, at "Civil Hospital" in Vittorio Veneto (TV), underwent TAPP laparoscopic inguinal hernia repair. The total hernias repaired were 362. RESULTS: The overall mean operative time was 45.58 min. (+/-15.2 min). All the procedures were performed in day surgery. There were no conversions to open repair or deaths in our series. We had 2 cases of small bowel occlusion and 5 relapsing hernias (1.3%), which we preferred to treat with an anterior approach. The mean follow-up was 30.4 (+/-5.6; range: 1 to 60 mo) months. No patients reported severe pain at 10 days, 7.2% (14 cases) reported mild pain at 3 months. There were no reports of night pain at 30 days. Approximately 90% of the patients had a return to physical-work capacity within 7 days, the remainder within 14 days. All patients were completely satisfied at 3 months. CONCLUSIONS: The analysis of the short-term postoperative outcomes of our experience enabled us to conclude that in the proper setting TAPP is feasible, effective, safe, and beneficial for patients and should be a routine part of any surgical practice providing adequate training has been undertaken and proper preparation observed. PMID- 19542859 TI - Impact of closed-suction drain in preperitoneal space on the incidence of seroma formation after laparoscopic total extraperitoneal inguinal hernia repair. AB - PURPOSE: To study the effect of closed suction drain in preperitoneal space on seroma formation after laparoscopic total extraperitoneal repair (TEP). METHODS: A closed suction drain-12G was put through the midline 5 mm port incision and removed with in 24 hours of operation in TEP repair. The results were compared with a group with TEP without any drain. RESULTS: In 929 patients (1753 hernias), drain was put in 849 patients (1607 hernias) and no drain was put in 80 patients (146 hernias). Follow-up ranged from 9 to 45 months. Seroma formation was significantly lower in the drain group (12/1607; 0.75%) compared with the nondrain group (22/146; 15.1%) (P<0.0001). Both the groups were comparable in pain scores, conversion to open, hospital stay, and days taken to return to normal activity and recurrence rates. There was no infection in either group. CONCLUSIONS: Drain after TEP significantly reduces the incidence of seroma formation without increasing the risk of infection or recurrence. PMID- 19542861 TI - Robotic camera holder as good as expert camera holder: a randomized crossover trial. AB - INTRODUCTION: This study aimed to compare the impact of robotic camera holder (RCH) and human camera holder (HCH) on product quality and procedure effectiveness of a simulated laparoscopic procedure. METHODS: This was a prospective randomized crossover trial including voluntary surgical residents. Block randomization generated RCH-HCH or HCH-RCH sequence allocation. The task was suturing a duodenal perforation on foam stomach with intracorporeally knot tying in a simulator. The camera was operated by the same robot and same expert. Product quality was measured by accuracy error, tissue damage, sliding knot, and leak. Procedure effectiveness was measured by operating time, nongoal directed actions, and dangerous actions. Kendall's coefficient tau_b was used for interrater reliability between 2 blinded assessors. RESULTS: Forty-four subjects performed their tasks as allocated. Product quality and procedure effectiveness were similar when first attempt of task was compared with the repeat task by same subject ignoring the type of camera holder. There was no evidence of significant unequal carryover effect when comparison was stratified by RCH-HCH or HCH-RCH sequences. There were no differences in product quality and procedure effectiveness when RCH was compared with HCH. Coefficient tau_b was > or = 0.80 for all but dangerous actions (0.72, P=0.08). CONCLUSIONS: RCH and HCH had similar impact on product quality and procedure effectiveness of simulated laparoscopic procedure. PMID- 19542862 TI - Is the health care system the next victim of the financial crisis? PMID- 19542863 TI - Association of abacavir and impaired endothelial function in treated and suppressed HIV-infected patients. AB - BACKGROUND: HIV-infected patients have accelerated atherosclerosis. Abacavir has been associated with increased risk of cardiovascular events, for reasons that remain to be elucidated. As endothelial dysfunction is central to the pathogenesis of atherosclerosis, we tested the hypothesis that current treatment with abacavir is associated with impaired endothelial function. METHODS: We studied a cohort of 61 antiretroviral-treated patients who had undetectable plasma HIV RNA levels. Endothelial function was assessed by measuring flow mediated dilation (FMD) of the brachial artery. We compared FMD in patients treated with or without abacavir, while adjusting for traditional risk factors and HIV-specific characteristics. RESULTS: The median age was 50 years (interquartile range 45-57). The median duration of HIV infection was 18 years, and the median CD4 cell count was 369 cells/microl. Thirty patients (49%) were receiving abacavir. Overall, the median FMD in the HIV-infected patients was low (3.5%; interquartile range 2.3-5.6%). The FMD was lower in the abacavir-treated patients than those not on abacavir (2.8 vs. 4.9%, P = 0.01). After adjustment for traditional risk factors, HIV-specific factors, and baseline brachial artery diameter, current abacavir use was independently associated with lower FMD (P = 0.017). Duration of therapy and CD4 cell count were not associated with reduced FMD. CONCLUSION: Endothelial function, a central mechanism in atherosclerosis and a marker of cardiovascular risk, is impaired among antiretroviral-treated patients with undetectable viral loads. Current use of abacavir was independently associated with impaired endothelial function. This finding suggests that abnormal endothelial function may underlie the clinically observed increased risk in myocardial infarction among abacavir-treated patients. PMID- 19542864 TI - Hepatitis C virus infections among HIV-infected men who have sex with men: an expanding epidemic. AB - BACKGROUND: Since 2000 outbreaks of sexually transmitted hepatitis C Virus (HCV) infections have been reported among HIV-infected men who have sex with men (MSM). We studied the prevalence and determinants of HCV-infection among MSM attending a large sexually transmitted infection (STI) clinic in the Netherlands. METHODS: In 2007-2008, 3125 attendees of the STI clinic Amsterdam, including 689 MSM, participated in an anonymous biannual crosssectional survey. Participants were interviewed and screened for HIV and HCV antibodies. Additionally, all anti-HCV positive and HIV-infected individuals were tested for HCV RNA. Using phylogenetic analysis, HCV strains of the STI clinic attendees were compared with those isolated from MSM with acute HCV in 2000-2007. Determinants of HCV-infection were analysed using logistic regression. RESULTS: Two of 532 (0.4%) HIV-negative MSM and 28 of 157 (17.8%) HIV-positive MSM were infected with HCV. Over the study period, HCV prevalence among HIV-infected MSM increased (14.6%-20.9%). Seven of 28 (25.0%) HIV/HCV coinfected MSM had acute HCV infection. Only five of 28 (17.9%) HIV/HCV coinfected MSM ever injected drugs (IDU). HIV-infection, IDU, fisting and gamma hydroxy butyrate (GHB)-use were significantly associated with HCV-infection. Phylogenetic analyses revealed a high degree of MSM-specific clustering. CONCLUSION: We found a high and increasing HCV prevalence in HIV infected MSM. Though not statistically significant, this trend, and the relatively large proportion of acute infections suggest ongoing transmission of HCV in HIV-positive MSM. Regardless of IDU, rough sexual techniques and use of recreational drugs were associated with HCV-infection; phylogenetic analysis supported sexual transmission. Targeted prevention, like raising awareness and routine testing, is needed to stop the further spread among HIV-infected MSM, and to prevent possible spillover to HIV-negative MSM. PMID- 19542865 TI - Tumor necrosis factor region polymorphisms are associated with AIDS and with cytomegalovirus retinitis. AB - BACKGROUND: The tumor necrosis factor (TNF) gene is located within the highly polymorphic major histocompatibility complex region, exhibiting the -308 GA promoter region polymorphism and six microsatellites (TNFa-f) spanning the region nearby the TNF locus. OBJECTIVE: In the present study, we evaluated the frequency of -308 GA and TNFa-e polymorphisms and respective haplotypes (in chromosomal sequence: TNFd-TNFe-308GA-TNFc-TNFa-TNFb), in 222 patients with AIDS, 52 of whom exhibited cytomegalovirus retinitis, and in 202 healthy HIV-negative individuals. METHOD: TNF microsatellite and single nucleotide polymorphism typings were performed by PCR followed by polyacrylamide gel electrophoresis. RESULTS: The TNF 308A allele and the 4-3-G-2-7-1 haplotype were associated with susceptibility to AIDS, whereas the TNFb4 allele and the 3-3-G-1-11-4 haplotype were associated with protection against AIDS development. The TNFc2 allele and the 4-1-G-2-2-1 haplotype, which contains the TNFc2 allele, were associated with cytomegalovirus retinitis. CONCLUSION: This study highlights that polymorphic sites spanning the region nearby the TNF locus are associated with AIDS per se and with cytomegalovirus retinitis in AIDS patients. PMID- 19542867 TI - Knowledge of HIV status, sexual risk behaviors and contraceptive need among people living with HIV in Kenya and Malawi. AB - BACKGROUND: Several studies support the need for effective interventions to reduce HIV transmission risk behaviors among people living with HIV/AIDS (PLWHAs). DESIGN: Cross-sectional nationally representative demographic health survey of Kenya (2003) and Malawi (2004-2005) that included HIV testing for consenting adults. METHODS: We analyzed demographic health survey data for awareness of HIV status and sexual behaviors of PLWHAs (Kenya: 412; Malawi: 664). The analysis was adjusted (weighted) for the design of the survey and the results are nationally representative. FINDINGS: Eighty-four percent of PLWHAs in Kenya and 86% in Malawi had sex in the past 12 months and in each country, 10% reported using condoms at last intercourse. Among sexually active PLWHAs, 86% in Kenya and 96% in Malawi reported their spouse or cohabiting partner as their most recent partner. In multivariate logistic regression models, married or cohabiting PLWHAs were significantly more likely to be sexually active and less likely to use condoms. Over 80% of PLWHAs were unaware of their HIV status. Of HIV-infected women, nearly three-quarters did not want more children either within the next 2 years or ever, but 32% in Kenya and 20% in Malawi were using contraception. INTERPRETATION: In 2003-2005, majority of PLWHAs in Kenya and Malawi were unaware of their HIV status and were sexually active, especially married or cohabiting PLWHAs. Of HIV-infected women not wanting more children, few used contraception. HIV testing should be expanded, prevention programs should target married or cohabiting couples and family planning services should be integrated with HIV services. PMID- 19542866 TI - Randomized, double-blind, placebo-matched, multicenter trial of abacavir/lamivudine or tenofovir/emtricitabine with lopinavir/ritonavir for initial HIV treatment. AB - BACKGROUND: Abacavir sulfate/lamivudine (ABC/3TC) and tenofovir DF/emtricitabine (TDF/FTC) are widely used nucleoside reverse transcriptase inhibitors for initial HIV-1 treatment. This is the first completed, randomized clinical trial to directly compare the efficacy, safety, and tolerability of these agents, each in combination with lopinavir/ritonavir in antiretroviral-naive patients. METHODS: Six hundred and eighty-eight antiretroviral-naive, HIV-1-infected patients were randomized in this double-blind, placebo-matched, multicenter, noninferiority study to receive a once-daily regimen of either ABC/3TC 600 mg/300 mg or TDF/FTC 300 mg/200 mg, both with lopinavir/ritonavir 800 mg/200 mg. Primary endpoints were the proportion of patients with HIV-1 RNA below 50 copies/ml at week 48 (missing = failure, switch included analysis) and the proportion of patients experiencing adverse events over 96 weeks. RESULTS: At week 48, 68% in the ABC/3TC group vs. 67% in the TDF/FTC group achieved an HIV-1 RNA below 50 copies/ml (intent-to-treat exposed missing = failure, 95% confidence interval on the difference -6.63 to 7.40, P = 0.913), demonstrating the noninferiority of ABC/3TC to TDF/FTC at week 48. Noninferiority of the two regimens was sustained at week 96 (60% vs. 58%, respectively, 95% confidence interval -5.41 to 9.32, P = 0.603). In addition, efficacy of both regimens was similar in patients with baseline HIV-1 RNA >or= 100 000 copies/ml or CD4 cell counts below 50 cells/microl. Median CD4 recovery (ABC/3TC vs. TDF/FTC, cells/microl) was +250 vs. +247 by week 96. Premature study discontinuation due to adverse events occurred in 6% of patients in both groups. Protocol-defined virologic failure occurred in 14% of patients in both groups. CONCLUSION: Both ABC/3TC and TDF/FTC provided comparable antiviral efficacy, safety, and tolerability when each was combined with lopinavir/ritonavir in treatment-naive patients. PMID- 19542868 TI - Intercalated duct lesions of salivary gland: a morphologic spectrum from hyperplasia to adenoma. AB - Intercalated duct lesions (IDLs) are rare, poorly understood and not well-studied lesions that have been associated with a small number of epithelial-myoepithelial carcinomas (EMC) and basal cell adenomas. To examine the nature of IDLs and their association with salivary gland tumors, we reviewed 34 lesions in 32 patients. The IDLs were stained with CK7, estrogen receptors (ER), progesterone receptors, lysozyme, S100, calponin, and CK14. The patients ranged in age from 19 to 80 years (mean 53.8) with a 1.7:1 female predominance. The majorities of IDLs were parotid lesions (82%), were small and nodular (average size 3.1 mm) and showed 3 architectural patterns: hyperplasia (20), adenoma (9), and hybrid forms (5). In 59% of cases, IDLs were seen in conjunction with another salivary gland tumor, most commonly basal cell adenoma (8 cases), followed by EMC (3 cases). One case showed a combination of intercalated duct hyperplasia and basal cell adenoma. The IDLs stained diffusely with CK7 (100%) and S100 (73%) and focally for ER (91%) and lysozyme (100%). Calponin and CK14 highlighted a thin myoepithelial cell layer around all ducts (100%). Normal intercalated ducts were also consistently positive for CK7 and lysozyme, and focally for ER, but were S100 negative. In summary, IDLs have a variety of patterns ranging from hyperplasia to adenoma with hybrid lesions and share morphologic and immunophenotypic features with normal intercalated ducts. There is an association with basal cell adenomas and EMC, which lends credence to their role as a putative precursor lesion. PMID- 19542869 TI - Complete hydatidiform mole with retained maternal chromosomes 6 and 11. AB - Distinction of hydatidiform moles from nonmolar specimens and their subclassification as complete hydatidiform mole (CHM) versus partial hydatidiform mole (PHM) are important for clinical practice and investigational studies to refine ascertainment of risk of persistent gestational trophoblastic disease which differs among these entities. Immunohistochemical analysis of p57 expression, a paternally imprinted maternally expressed gene on 11p15.5, and molecular genotyping are useful for improving diagnosis. CHMs are characterized by androgenetic diploidy, with the loss of p57 expression owing to lack of maternal DNA. Loss of p57 expression distinguishes CHMs from both PHMs (diandric triploidy) and nonmolar specimens (biparental diploidy) which retain expression. In the process of evaluating molar specimens in our laboratory with p57 immunohistochemistry and molecular genotyping, we identified a morphologically typical androgenetic diploid CHM with aberrant diffuse p57 expression. Molecular genotyping by short tandem repeat markers and genome-wide copy number analysis by single nucleotide polymorphism array established androgenetic diploidy with retained maternal copies of chromosomes 6 and 11, with aberrant p57 expression attributable to the latter. This case, only the second reported to date, illustrates the value of combined traditional pathologic and ancillary molecular techniques for refined diagnosis of molar specimens. Specimens with morphologic features suggestive of CHM yet retaining p57 expression should be subjected to molecular genotyping to establish a definitive diagnosis because misclassification as PHM underestimates the risk of persistent gestational trophoblastic disease. We recommend use of p57 immunohistochemistry and molecular genotyping to evaluate all products of conception specimens for which there is any consideration of a diagnosis of hydatidiform mole. Genome-wide analysis has the potential to assist in localizing imprinted genes critical for determining the morphologic and behavioral phenotypes of hydatidiform moles. PMID- 19542870 TI - Pathogenetic pathways in ovarian endometrioid adenocarcinoma: a molecular study of 29 cases. AB - It has been recently suggested that ovarian serous carcinoma follows a dualistic pathway with low-grade carcinomas arising from borderline tumors and high-grade carcinomas originating de novo. Similarly, our group has shown that based on their molecular profile endometrioid borderline tumors could predate low-grade endometrioid ovarian carcinomas (EOC). It is not clearly understood if low-grade EOC is in turn related to high-grade EOC, or if high-grade EOC may also arise de novo. The aim of our study was to compare the molecular profile of grade 1, 2, and 3 EOCs. Twenty-nine EOCs were selected including 10 grade 1 (G1), 11 grade 2 (G2), and 8 grade 3 (G3). Selected blocks were immunostained with beta-catenin and p53, and also microdissected, DNA extracted and amplified by polymerase chain reaction with primers for exon 3 of the beta-catenin gene, codons 12 and 13 of KRAS and codons 1 to 9 of PTEN. The length of BAT-26 and BAT-25 was analyzed to determine microsatellite instability (MSI). Patients with G1 EOC ranged from 21 to 71 (mean 52) years, those with G2 tumors ranged from 43 to 66 (mean 56) years, and patients with G3 EOC ranged from 41 to 67 (mean 57) years. Immunohistochemical analysis for beta-catenin showed nuclear staining in 14 cases (7 G1, 5 G2, and 2 G3 tumors), whereas the rest showed membranous staining. Beta catenin mutations were found in only 3 G1 tumors. KRAS mutations were seen in 5 EOCs (2 G1 and 3 G2). MSI and mutations of PTEN were both detected in 1 G1 and 1 G2 tumor, respectively. There was no overlapping expression of MSI, beta-catenin, PTEN, or KRAS mutations. Finally, p53 overexpression was present in 6 EOCs (5 G3 and 1 G2), all G3 p53 positive tumors being negative for all other markers, whereas the G2 tumor also showed a KRAS mutation. In conclusion, beta-catenin and KRAS mutations, and MSI were strongly associated with low-grade EOC. In contrast, p53 overexpression characterized high-grade EOC, with no other molecular alterations present in the vast majority of these tumors. On the basis of these results, we suggest that there may also be a dual pathogenetic pathway for EOC. PMID- 19542871 TI - Invasive urothelial carcinoma with chordoid features: a report of 12 distinct cases characterized by prominent myxoid stroma and cordlike epithelial architecture. AB - Urothelial carcinoma is morphologically heterogeneous and many variant forms have been described. We have encountered several invasive urothelial carcinomas with a unique chordoid morphology characterized by prominent cellular cording and associated myxoid stromal matrix, a pattern closely resembling extraskeletal myxoid chondrosarcoma. This morphologic appearance, to our knowledge, has not been formally described in urothelial carcinoma. A series of 166 consecutive invasive urothelial carcinomas were reviewed to identify cases with cellular cording and myxoid stroma. The patient age, sex, tumor stage, morphologic features, association with typical urothelial carcinoma, and clinical outcome were recorded. Immunostains for p63, cytokeratin (CK) 34BE12, CK20, calponin, glial fibrillary acidic protein, S-100 protein, oncofetal protein glypican-3, and brachyury were performed on 7 cases. Mucin histochemistry was performed on 8 cases to evaluate the extracellular myxoid material. Eleven of the 166 (7%) consecutive invasive urothelial carcinomas had areas with a chordoid appearance. A total of 12 cases were analyzed including the addition of a consult case. The patients' ages ranged from 50 to 85 years (mean: 68 y); there were 8 males and 4 females. The specimens consisted of 5 cystectomies, 6 transurethral resections, and 1 anterior exenteration with right nephroureterectomy. Morphologically, each case had at least focal areas in which acellular myxoid stroma was associated with the carcinoma cells. When well developed, the neoplastic cells had scant eosinophilic cytoplasm and were arranged into cords closely mimicking extraskeletal myxoid chondrosarcoma, chordoma, mixed tumor/myoepithelioma of soft tissue, and yolk sac tumor. The percentage of tumor with a chordoid appearance ranged from 5% to 95% (mean: 39%; median: 25%). No conventional sarcomatous differentiation, no intracytoplasmic mucin, and no glandular formation were present in any case. All 12 cases had foci of typical urothelial carcinoma present at least focally and a gradual transition to the chordoid pattern was commonly seen. Immunophenotypically, all 7 cases evaluated showed strong immunoreactivity for p63 (nuclear) and CK34BE12 (cytoplasmic). Immunostains for CK20, calponin, glial fibrillary acidic protein, oncofetal protein glypican-3, and brachyury and were negative in the 7 cases studied (0 out of 7), whereas S 100 protein had focal staining (<5%) in 1 case. The myxoid stromal component was diffusely colloidal iron and Alcian blue positive in all 8 cases examined; periodic acid Schiff was negative in all 8 cases, whereas mucicarmine was only focally positive in 2 of 8 cases. Most cases were high stage (pT4: 5, pT3: 4, pT2: 2, and pT1: 1), and 6 of 8 cases (75%) with nodal sampling had metastatic disease. In 1 case, the lymph node metastasis had areas with chordoid morphology. Nine of 12 patients had available follow-up: 2 were dead of disease (1 and 10 mo), 4 were alive with disease (5 to 8 mo) with distant metastasis in 3, and 3 had no evidence of disease at last follow-up (2 to 120 mo). In summary, we describe a morphologic pattern of urothelial carcinoma with a distinct chordoid appearance that may potentially mimic a spectrum of primary vesical and nonvesical neoplasms with myxoid or mucinous components. These carcinomas maintain an immunophenotype characteristic of urothelial carcinoma and usually present with high stage disease. PMID- 19542872 TI - Uterine tumors resembling ovarian sex cord tumors (UTROSCT) lack the JAZF1-JJAZ1 translocation frequently seen in endometrial stromal tumors. AB - Uterine tumor resembling ovarian sex cord tumor (UTROSCT) is a rare uterine neoplasm composed predominantly or exclusively of cells which resemble those seen in sex cord tumors of the ovary. Since its initial morphologic description, it has been unclear whether UTROSCT represents a variant within the spectrum of endometrial stromal tumors (ESTs), which may rarely exhibit areas of sex cord like differentiation, or whether it is a distinct uterine neoplasm unrelated to ESTs. Recently, several studies have revealed a recurrent t(7;17) translocation resulting in a JAZF1-JJAZ1 gene fusion in over 60% of EST and its variants, including 2 out of 4 endometrial stromal tumors with sex cord-like elements (ESTSCLE). We examined UTROSCTs for evidence of the JAZF1-JJAZ1 gene fusion by fluorescence in situ hybridization and by reverse transcriptase polymerase chain reaction in 24 and 20 cases, respectively. The JAZF1-JJAZ1 gene fusion was not identified in any tumor by either method. Although we cannot entirely exclude that UTROSCT represents a variant of ESTSCLE which lacks this translocation, our findings suggest that UTROSCT does not share the genetic mechanism common to the majority of ESTs with or without sex cord-like differentiation, and therefore most likely represents a distinct neoplasm unrelated to ESTSCLE. PMID- 19542873 TI - Mycophenolic acid (cellcept and myofortic) induced injury of the upper GI tract. AB - BACKGROUND: Mycophenolic acid (MPA) is an immunosuppressant drug commonly used in patients undergoing solid organ transplant. Although its pattern of inducing injury in the colon is well-known and features prominent crypt apoptosis that mimics graft-versus-host-disease, the injury pattern in the upper gastrointestinal (GI) tract is less extensively documented. We studied the pattern of upper GI tract injury in symptomatic patients taking MPA. DESIGN: Twenty solid organ transplant patients who were taking MPA and had concurrent upper GI tract biopsies were identified on a laboratory information system search. From these 20 patients, 17 duodenal, 18 gastric, and 7 esophageal biopsies were examined. All patients were symptomatic. Apoptosis and patterns of chronic and active injuries were assessed on standard hematoxylin and eosin, periodic acid-Schiff/Alcian blue, and Diff-Quik stained slides. To measure the significance of apoptosis, we standardized the apoptotic counts in normal biopsies using duodenal, gastric, and esophageal biopsies from 45 normal cases and performed statistical analysis. For the purposes of this study, we regarded apoptotic counts higher than the mean plus 2 SDs as significant. Thus the cut-off values for apoptosis were > or =2 apoptotic bodies/100 crypts for duodenum, > or =3/100 glands for stomach, and > or =2/10 high-power field for esophagus. RESULTS: GI-related symptoms and abnormalities manifested between 1 month and 10 years posttransplant and included diarrhea (55%); nausea (45%); abdominal pain (35%); vomiting (25%); GI bleeding (15%); dysphagia (10%); dyspepsia, anemia, and hematemesis (5% for each). Most (14 out of 17, 82%) duodenal biopsies showed apoptotic counts of > or =2/100 crypts, 28% (5 out of 18) of gastric biopsies showed apoptotic counts of > or =3/100 glands, and 57% (4 out of 7) of esophageal biopsies showed apoptotic counts of > or =2/10 high-power field. Four gastric biopsies showed a previously undescribed injury pattern of parietal cells resembling the ballooning degeneration. Additional pathologic findings included: chronic peptic duodenitis (6 out of 17, 35%), active duodenitis (1 out of 17, 6%), and celiac-like features (2 out of 17, 12%) in the duodenum; chemical gastropathy (3 out of 18, 17%), active chronic gastritis without Helicobacter pylori (2 out of 18, 11%), and erosion (1/18,6%) in the stomach; reactive epithelial change (3 out of 7, 43%), active esophagitis (3 out of 7, 43%), ulceration (2 out of 7, 29%), and erosion (1 out of 7, 14%) in the esophagus. Serum MPA levels were available in 7 patients, 6 of whom had abnormal duodenal apoptotic counts. On follow-up, available for 16 patients, symptoms improved in all the patients whose dose was decreased or whose medication was withdrawn (10 out of 10) and symptoms persisted in all the patients whose dose was not altered (6 out of 6). Follow-up biopsies after reduction of the medication dose were available for 1 patient and showed substantial reduction in apoptosis. In contrast, follow-up biopsies from 1 patient whose dosage was not altered showed persistent abnormal apoptotic counts in the duodenum. CONCLUSIONS: As noted by others (Parfitt JR, Jayakumar S, Driman DK. Am J Surg Pathol. 2008; 32:1367 1372), mycophenolate mofetil-associated injury of the upper GI tract, like that in the colon, is characterized by prominent apoptosis similar to that of mild or grade I graft-versus-host-disease injury. We offer apoptotic count guidelines, which we hope will facilitate recognition of mycophenolate mofetil-associated injury in the upper GI tract. PMID- 19542874 TI - Clusterin is expressed in normal synoviocytes and in tenosynovial giant cell tumors of localized and diffuse types: diagnostic and histogenetic implications. AB - Tenosynovial giant cell tumors arise from synovium of joints, bursae, or tendon sheaths, and are classified into localized and diffuse types based on the growth pattern and clinical behavior. The mononuclear component of these tumors includes small histiocytoid cells and large mononuclear cells, which are positive for desmin in about 50% of cases. This study seeks to further characterize the immunophenotype of these tumors, and investigates the utility of clusterin as a diagnostic marker. Immunostaining for clusterin was performed on 40 cases of tenosynovial giant cell tumor (11 localized and 29 diffuse). Most cases were also stained for desmin, CD163, CD21, and CD35. Four cases were stained for podoplanin/D2-40 and CXCL13. Clusterin staining was diffuse and strong in the large mononuclear cells in all cases. Desmin positivity in the large cells was identified in 24 out of 34 cases (71%), but was seen in only a subset of cells (<5% to 80%), with 19 out of 24 cases (79%) showing positivity in 10% or less. The large cells were positive for podoplanin in 4 out of 4 cases, but negative for CD163, CD21, CD35, and CXCL13. The smaller histiocytoid cells were positive for CD163 and negative for all other markers. When present, non-neoplastic synoviocytes were positive for clusterin and podoplanin, and focally positive for desmin. Clusterin is a highly sensitive marker for tenosynovial giant cell tumors, which has diagnostic utility in challenging cases. The observed staining patterns provide evidence linking the large mononuclear cells with normal synoviocytes and support that tenosynovial giant cell tumors are neoplasms showing synovial differentiation. PMID- 19542875 TI - Arrhythmia and acute coronary syndrome suppression and cardiac resuscitation management with bretylium. AB - It is well known that electric shock can both initiate and terminate ventricular fibrillation. Refractory ventricular fibrillation (RVF) may often be an iatrogenic paradoxical result of early, frequent, excessive salvos of DC current countershocks and inappropriate off-label drug use, particularly aggressive epinephrine administration. Evidence suggests that the current advanced cardiac life support pharmacology protocol for cardiac resuscitation may contribute to disappointing survival in patients with out-of-hospital cardiac arrest. Controlled studies and new theoretical consideration suggest the protocol may induce RVF. In contrast, studies suggest that immediate adequate intravenous bretylium administration therapy together with sustained effective chest compressions can induce chemical defibrillation or facilitate electrical defibrillation as well as reduce the intensity, or even need for potentially heart-damaging countershock, where early frequent excessive current shocks are likely to increase refractory arrhythmia as demonstrated in animals and in humans. Salvos of shocks do not allow time between shocks for uniform recovery of normal electrical parameters needed to restore a stable heart rhythm. This may occur by inadvertently administering shock during the vulnerable period of the cardiac cycle. There are compelling existing data to demonstrate that bretylium and cardiopulmonary resuscitation (CPR) delivered before initiating shock therapy is likely to provide the best outcome in cardiac arrest. But, most importantly, adequate CPR chest compressions administered while bretylium is being infused also provide the opportunity to wash out electrically destabilizing electrolytes that have leaked from or are abnormally transported by functionally damaged membranes of fibrillation-induced ischemic myocytes. This may cause abnormal compartment redistribution of electrolytes that may facilitate RVF by heterogeneously partially depolarizing ischemic myocytes. Although efforts have been made to provide hard science for advanced life support, the guidelines are a product of consensus, the give and take of collegiality and intuition rather than rigorous controlled studies. Bretylium has a direct antifibrillatory action normalizing myocyte membrane currents, which restores intracompartmental normal electrolyte balance. In addition, adrenergic blockade by bretylium dilates coronary arteries, increasing effective O2 delivery by CPR. The free and aggressive use of epinephrine is toxic. Catechalomines cause coronary spasm and puts myocardial metabolism into damaging hypermetabolic overdrive to support the "fight or flight reflex" rapidly depleting adenosine triphosphate needed for cardiac electrical and mechanical recovery. Moreover, the value of epinephrine to resuscitation has never been demonstrated in a controlled human study, whereas its potential damage has been largely ignored. Epinephrine's potential deleterious actions that might compromise resuscitation are well established and reviewed here. PMID- 19542877 TI - Superior and superomedial pedicle wise-pattern reduction mammaplasty: maximizing cosmesis and minimizing complications. AB - Our center has refined a wise-pattern superior and superomedial pedicle breast reduction technique with predictable quality outcomes giving high patient satisfaction and a very low complication or revision rate. Fifty patients who underwent bilateral superior or superomedial pedicle wise-pattern breast reduction were included in the study. Demographic and perioperative data were collected, as were details of complications. A 21-item postal questionnaire was used to evaluate satisfaction with outcome parameters. The superomedial pedicle technique was used in 86% reductions, and the superior pedicle in 14%, and the mean weight removed from the breasts was 778 g (range, 244-1766 g). Mean patient satisfaction with outcome parameters, as well as maintenance of shape was excellent, as were mean improvements in symptoms of macromastia. No patient required a revision procedure. Superior and superomedial pedicle wise-pattern scar reduction mammaplasty with the technique presented predictably gives excellent results and patient satisfaction, with long-term preservation of shape. PMID- 19542876 TI - Temporomandibular joint dislocation reduction technique: a new external method vs. the traditional. AB - The traditional intraoral approach for temporomandibular joint dislocations reduction, although effective, has some disadvantages. Here, a new extraoral approach is described. This study was performed to evaluate this new method's success rate. Patients visiting an emergency room were randomly allocated to 2 groups; one group was reduced with the extraoral approach and the other with the intraoral method. Among 29 attempts with the conventional method, 25 were successful (86.2%; 95% confidence interval: 73-100) and among 29 attempts with the external method, 16 were successful (55.2%; 95% confidence interval: 39-79). This difference was statistically significant. Because of the benefits of the external approach, such as avoiding hand bites and disease transfer, it can be a reasonable choice to reduce a dislocated temporomandibular joint. PMID- 19542878 TI - Donor site sensitivity after breast reconstruction with deep inferior epigastric artery perforator flap. AB - The aim of this study was to examine pressure sensitivity at the donor site after breast reconstruction with deep inferior epigastric artery perforator (DIEAP). In a cross-sectional survey, 2 groups of patients were analyzed. The DIEAP group consisted of 30 women who had previously had secondary breast reconstruction with DIEAP flap after mastectomy for breast cancer. The control group consisted of 7 women with no previous abdominal incisions planned for secondary breast reconstruction with DIEAP. Pressure thresholds were tested within the margins of the abdominal wall using Semmes-Weinstein monofilaments. In the DIEAP group a pattern of higher pressure thresholds was observed in the proximity of the scar. Comparing the 2 groups, significant higher pressure thresholds were found in the DIEAP group in the scar on both sides and in the midline from the scar to the umbilical level. Our data show that the abdominoplasty performed during breast reconstruction with DIEAP reduces cutaneous sensitivity in the donor site area. PMID- 19542879 TI - A review of intraoperative airway management for midface facial bone fracture patients. AB - In craniofacial trauma patients, oral route endotracheal intubation may thwart the accuracy of dental occlusion and nasotracheal intubation carries the risk of intracranial invasion in skull base fracture cases. Between November 2005 and June 2006, patients receiving facial bone fracture operations at Kaohsiung Medical University Hospital were enrolled in this study. Intraoperatively, the endotracheal tube was pushed to either the retromolar space or the missing tooth space and secured by two 4.0 silk stitches. Then, surgeons could perform the usual procedure to explore the fracture sites, check the occlusion and correct the deviated nose without limitation. Also, for better understanding the time needed for various intubation techniques, a time-measuring study was performed. Ninety-one patients were treated by this method. Most of them were satisfied with the result of occlusion and nasal contour. Only 2 patients received second surgery to correct nasal deformity. One hundred seventeen anesthesia procedures were checked. In average, an experienced anesthesiologist could successfully intubate a patient in less than 105 seconds. The advantages and reported complications of different intubation methods were discussed. This retromolar position and tooth fixation technique allowed surgeons to correct the dental occlusion and nasal deformity simultaneously. It has served well for zygoma fracture, maxilla fracture, and Le Fort II fracture patients. It is worthy of consideration in management of middle face trauma patients involving occlusion change and nasal deviation. PMID- 19542880 TI - Renal venous thrombosis in a newborn with prothrombotic risk factors. AB - Renal venous thrombosis (RVT) is a rare but a well recognized entity in children and neonates. The clinical signs of neonatal RVT include hypertension, enlarged kidney(s), hematuria, renal insufficiency, proteinuria, thrombocytopenia, or all. Persisting impairment of kidney function and hypertension are serious and common complications in patients with RVT. Risk factors for the development of RVT include maternal diabetes mellitus, pathologic states associated with thrombosis (e.g., shock, dehydration, perinatal asphyxia, polycythemia), and sepsis. Inherited prothrombotic abnormalities have been described in some reports of RVT. We report the case of a male newborn with left RVT and associated homozygosity for both factor V Leiden (G1691A) and methylenetetrahydrofolate reductase C677T mutations in addition to elevated serum lipoprotein (a). The patient was treated with heparin. We believe our case to be the first reported case in the English medical literature of such an association between neonatal RVT and homozygosity for both factor V Leiden and methylenetetrahydrofolate reductase. This case and other studies clearly demonstrate that neonatal RVT should be evaluated for thrombophilia conditions. PMID- 19542881 TI - Gene therapy for hemophilia A. Friend or foe? PMID- 19542882 TI - The clinical importance of laboratory-defined aspirin resistance in patients presenting with non-ST elevation acute coronary syndromes. AB - In this study, we aimed to assess the factors associated with laboratory-defined aspirin resistance and the relationship of this laboratory-defined aspirin resistance with thrombolysis in myocardial infarction risk score, markers of cardiac necrosis, and inflammatory and thrombotic risk factors in patients with unstable angina or non-ST elevation myocardial infarction. Ninety-seven patients who were under aspirin therapy and hospitalized with unstable angina/non-ST elevation myocardial infarction were included in the study. Laboratory-defined aspirin sensitive and resistant groups were determined by platelet function analyzer; aspirin resistance was defined as collagen/epinephrine closure time less than 165 s. Laboratory-defined aspirin resistance was noted in 29 patients (29.9%), and non-ST elevation myocardial infarction was observed in 46 patients (47.4%). Patients in the group with laboratory-defined aspirin resistance had significantly higher thrombolysis in myocardial infarction risk scores (P < 0.001). When the details of cardiac myonecrosis markers were compared, baseline and follow-up creatine kinase-myocardial band and troponin I values were higher in laboratory-defined aspirin-resistant group. Multivariate analyses revealed that laboratory-defined aspirin resistance was an independent predictor of non-ST elevation myocardial infarction (P = 0.022). Laboratory-defined aspirin resistance is associated with non-ST elevation myocardial infarction, higher markers of cardiac necrosis and thrombolysis in myocardial infarction risk score in patients hospitalized with unstable angina/non-ST elevation myocardial infarction. PMID- 19542883 TI - Cardiorenal syndromes. AB - PURPOSE OF REVIEW: The purpose of the present review is to identify the mechanisms involved in the syndrome related to combined heart-kidney dysfunction. RECENT FINDINGS: The bidirectionality of the syndrome and the various time frames involved in the different clinical pictures have induced to generate a new definition of the cardiorenal syndrome, focusing on five specific subtypes (acute cardiorenal syndrome, type 1; chronic cardiorenal syndrome, type 2; acute renocardiac syndrome, type 3; chronic renocardiac syndrome, type 4; and secondary cardiorenal syndrome, type 5). SUMMARY: The new definition allows to characterize the various clinical scenarios and to identify patients with different subtypes in which the primary disorder and the sequelae are clearly described. Biomarkers for early diagnosis of heart and kidney dysfunction may further contribute to a clearer definition of the disorder. The new classification will allow to test diagnostic tools and prevention strategies as well as therapeutic measures that in the past might not have been applied properly due to the lack of a consistent classification of the syndrome. PMID- 19542884 TI - Vasoactive drugs for vasodilatory shock in ICU. AB - PURPOSE OF REVIEW: Vasoactive drugs are the mainstay of hemodynamic management of vasodilatory shock when fluids fail to restore adequate tissue perfusion. In this review, studies published during the past year that increase our understanding of the use of vasoactive drugs in the ICU are discussed. RECENT FINDINGS: The Vasopressin and Septic Shock Trial did not find a difference between low-dose vasopressin and norepinephrine vs. norepinephrine alone in the hemodynamic support of septic shock, suggesting that either approach is reasonable. However, vasopressin may be beneficial in the less severe septic shock subgroup. In this study, patients who were also treated with corticosteroids, vasopressin, compared with norepinephrine, were associated with significantly decreased mortality. Epinephrine, phenylephrine and terlipressin can be used safely in the ICU setting as first-line therapy for septic shock. The incidence of global left ventricular hypokinesia in patients with septic shock is 60%, much higher than previously described. Although dobutamine remains the gold standard therapy for septic myocardial depression, combined milrinone and metoprolol therapy may be an effective alternative therapy. SUMMARY: Current evidence does not support a clear recommendation of one vasopressor over another; indeed norepinephrine, vasopressin, terlipressin, phenylephrine and epinephrine may be used safely with similar survival outcomes. PMID- 19542885 TI - Respiratory complications after major surgery. AB - PURPOSE OF REVIEW: To discuss the recent literature concerning the significance of respiratory complications as a determining factor in postoperative complications after major surgery. Although many studies have identified risk factors focusing on the prevention of respiratory complications, these complications continue to be a significant cause of morbidity and mortality. Here, we will examine the diagnosis, contributing factors, consequences, and early treatment of respiratory complications. RECENT FINDINGS: General anesthesia and surgery are the main causes of postoperative respiratory complications. Atelectasis, a common respiratory complication, may contribute to pneumonia and acute respiratory failure. Recently, it has been shown that activation of abdominal muscles during the induction of anesthesia contributes to a reduction of lung capacity, leading to a higher degree of atelectasis. Additionally, long term mortality at 5 and 10 years has recently been shown to remain significantly increased in patients with respiratory complications. Prevention or early therapy of respiratory complications may, therefore, be beneficial in improving outcome in postoperative patients. SUMMARY: Postoperative respiratory complications may have significant deleterious consequences. Increasing the understanding of the underlying causes of respiratory complications and developing early treatment strategies will likely provide improved benefits. To date, early treatment with prophylactic or therapeutic continuous positive airway pressure has proved beneficial in an abdominal surgical patient population; however, the efficacy in a general population remains unclear. PMID- 19542886 TI - Psychogenic movement disorders. AB - PURPOSE OF REVIEW: This review summarizes the progress made in the area of psychogenic movement disorders (PMDs) over the past 2 years, and a simplified classification of diagnostic certainty is proposed that incorporates electrophysiological assessment. RECENT FINDINGS: Functional magnetic resonance imaging studies have demonstrated altered blood flow in conversion disorders that may reflect changes in synaptic activity. Electrophysiological testing shows limitations in distinguishing between psychogenic and organic propriospinal myoclonus and dystonia. Recent evidence cautions against the uncritical acceptance of all cases of posttraumatic myoclonus and 'jumpy stump' as being organic in nature. 'Essential palatal tremor' is recognized as a rather heterogeneous group of tremors that includes psychogenic tremor. Two recent studies evaluating the long-term prognosis of psychogenic tremor differ in the degree of unfavorable outcome. Different groups of PMDs might have distinctive gait characteristics with prognostic, diagnostic, or therapeutic value. Two recent reviews provide comprehensive information on the understudied area of PMDs in children. SUMMARY: The diagnosis of PMDs should not be regarded as a diagnosis of exclusion. Careful clinical assessment is critical, and imaging or electrophysiological studies may provide important insights and confirmation of the diagnosis though some cases remain challenging and current assessments fail to provide needed clarification. Treatment is often delayed, contributing to a largely unfavorable long-term outcome. Well designed randomized control trials that validate and compare therapeutic options are urgently required. PMID- 19542887 TI - Genetic dissection of the mouse CNS using magnetic resonance microscopy. AB - PURPOSE OF REVIEW: Advances in magnetic resonance microscopy (MRM) make it practical to map gene variants responsible for structural variation in brains of many species, including mice and humans. We review results of a systematic genetic analysis of MRM data using as a case study a family of well characterized lines of mice. RECENT ADVANCES: MRM has matured to the point that we can generate high contrast, high-resolution images even for species as small as a mouse, with a brain merely 1/3000th the size of humans. We generated 21.5-micron data sets for a diverse panel of BXD mouse strains to gauge the extent of genetic variation, and as a prelude to comprehensive genetic and genomic analyses. Here we review MRM capabilities and image segmentation methods; heritability of brain variation; covariation of the sizes of brain regions; and correlations between MRM and classical histological data sets. SUMMARY: The combination of high throughput MRM and genomics will improve our understanding of the genetic basis of structure-function correlations. Sophisticated mouse models will be critical in converting correlations into mechanisms and in determining genetic and epigenetic causes of differences in disease susceptibility. PMID- 19542888 TI - Transplantation tolerance in an age of induced pluripotency. AB - PURPOSE OF REVIEW: In a seminal paper in 2006, Yamanaka and coworkers reported the reprogramming of terminally differentiated murine cells to a pluripotent state, largely indistinguishable from conventional embryonic stem cells. The introduction of defined transcription factors via retroviral transduction revealed, in principle, how pluripotency, once thought to be lost at an early stage of embryogenesis, could be reawakened in adulthood. RECENT FINDINGS: Since these initial findings, induced pluripotency has been reported using human as well as mouse cells and through the introduction of recombinant proteins, thereby avoiding the use of retroviruses for genetic modification. This approach, therefore, harnesses the traditional plasticity of embryonic stem cells as a source of therapeutic cell types and tissues, but without the many ethical issues with which they have become synonymous. The potential for exploiting pluripotency in this way also raises the tantalizing prospect of personalized therapies, spawning a new chapter in the story of regenerative medicine. SUMMARY: Given the promise of induced pluripotency and the disarming ease with which it can be achieved, it is perhaps timely to address the likely effect it will have on the field of transplantation and to ask whether the generation of induced pluripotent stem cells of autologous origin is ever likely to render redundant the need for transplantation tolerance. PMID- 19542889 TI - Innate immunity in heart transplantation. AB - PURPOSE OF REVIEW: Cardiac transplantation is the treatment of choice for end stage heart failure, but its efficacy is limited by the development of cardiac allograft vasculopathy (CAV). Although the adaptive immune system is efficiently suppressed by conventional drugs, the innate immune system is largely unaffected. The innate response may contribute both to stimulation of the adaptive response and to the future development of CAV. RECENT FINDINGS: Stimulation of Toll-like receptors by endogenous ligands released in response to ischemia/reperfusion causes an inflammatory milieu favorable to graft rejection and unfavorable to tolerance. New evidence suggests that natural killer cells have previously unknown memory-like features and are capable of graft rejection. Their role in rejecting the cardiac allograft has previously been underestimated. Complement deposition may also contribute to acute cellular rejection and CAV. SUMMARY: The innate immune system is an important but neglected component of allograft rejection. Drugs that target Toll-like receptors, natural killer cells and complement may play an important role in preventing CAV and achieving tolerance to cardiac allografts. PMID- 19542890 TI - Renal-sparing strategies in cardiac transplantation. AB - PURPOSE OF REVIEW: Renal dysfunction due to calcineurin inhibitor (CNI) toxicity is a major clinical problem in cardiac transplantation. The aim of the article is to review the efficacy and safety of various renal sparing strategies in cardiac transplantation. RECENT FINDINGS: Small studies have documented that late initiation of CNI is safe in patients treated with induction therapy at the time of transplantation. Use of mycophenolate is superior when compared with azathioprine to allow for CNI reduction. More substantial reduction in CNI levels is safe and effective with the introduction of sirolimus or everolimus. However, studies that use very early CNI discontinuation have found an increased risk of allograft rejection, and this strategy requires further study before it can be routinely recommended. CNI discontinuation late after cardiac transplantation seems more effective than CNI reduction in terms of preserving renal function. Patients with longstanding CNI treatment or proteinuria are less likely to respond favourably to a switch from a CNI-based regimen to a proliferation signal inhibitor-based regimen. SUMMARY: Each cardiac transplant recipient with renal dysfunction must be individually evaluated with respect to degree of renal dysfunction, proteinuria and rejection risk and a renal sparing strategy chosen accordingly. PMID- 19542891 TI - New-onset diabetes after transplantation: a review of recent literature. AB - PURPOSE OF REVIEW: New-onset diabetes after transplantation (NODAT) is a serious complication of organ transplantation. Data from kidney transplant studies show that NODAT is a strong independent predictor of graft failure and cardiovascular mortality. This article reviews NODAT in context of some of the recent data on definition, incidence, risk factors, genetics, and the impact on graft survival and cardiovascular events. RECENT FINDINGS: The reported incidence of NODAT continues to be high. The variability in the incidence can be attributed to varying definitions used in studies and also to the immunosuppressive regimens used at various centers. A 5-day oral glucose tolerance test may be a better predictor for developing NODAT. Comparison studies of various immunosuppressants in contributing to this condition show variable and conflicting results. Hepatitis C has emerged as a strongly associated risk factor and sirolimus may not be less diabetogenic, as thought before. In addition to serious infections, NODAT has been associated with increased cardiovascular risk and atherosclerosis and higher graft failures. SUMMARY: New-onset diabetes continues to be a common and potentially serious complication after organ transplantation. Risk stratification, early diagnosis, and intervention for this condition may contribute to better long-term graft survival and help in reducing cardiovascular mortality. PMID- 19542892 TI - Cardiac imaging in myocardial sarcoidosis and other cardiomyopathies. AB - PURPOSE OF REVIEW: The clinical role of emerging imaging technologies for diagnosing cardiac sarcoidosis and other cardiomyopathies is evolving. An up-to date review of the role of various imaging modalities in the evaluation of cardiac sarcoidosis and other cardiomyopathies is presented. RECENT FINDINGS: No study prospectively established the accuracy of each of the various techniques for diagnosing myocardial involvement in patients with suspected cardiac sarcoidosis. Cardiac magnetic resonance imaging (CMR) is demonstrated to have a sensitivity of 100% and specificity of approximately 80%, and positive predictive value of approximately 55% in diagnosing cardiac sarcoidosis. Recent studies have shown that 18F-2-fluoro-2-deoxyglucose positron emission tomography (FDG-PET) has 100% sensitivity of detecting earlier stages of sarcoidosis. Both the FDG-PET and CMR may provide complementary information for the diagnosis and assessment of efficacy of therapy in patients with cardiac involvement from sarcoidosis. SUMMARY: Clinical and subclinical cardiac involvement is common among patients with sarcoidosis. A structured clinical assessment incorporating advanced cardiac imaging with CMR and FDG-PET scanning is more sensitive than the established clinical criteria. CMR is an established imaging modality in the diagnosis of various other cardiomyopathies. Well designed prospective clinical trials are awaited to define the exact role of these imaging studies in the diagnosis and guidance of therapy. PMID- 19542893 TI - Seasonal variability of venous thromboembolism. AB - PURPOSE OF REVIEW: Seasonal variations in the incidence of cardiovascular disease, mainly characterized by a winter peak, have been consistently reported. Some evidence now also exists on potential seasonal variations in the incidence of venous thromboembolism (VTE). Of interest, seasonal variability in the incidence of deep vein thrombosis and fatal and nonfatal pulmonary embolism appears to differ; however, the results of available studies are not unequivocal. This review will briefly summarize current evidence in this area. RECENT FINDINGS: Recent studies indicate a seasonal variability in the incidence of VTE, with a pattern that is independent of sex, age, type of event, or underlying risk factors. Usually, these studies found a higher VTE incidence during the winter months and a lower incidence in the summer months. The exact mechanism of this variability is not completely understood, but it may be at least in part explained by changes in coagulation factor levels. SUMMARY: Confirmation of these findings and a better understanding of underlying mechanisms could help physicians to identify patients or periods with increased risk of VTE in order to further improve current prophylactic strategies. PMID- 19542894 TI - Home blood pressure monitoring in children and adolescents: a systematic review. AB - OBJECTIVE: As in the adults, in children and adolescents with elevated blood pressure (BP), the conventional office BP measurements might lead to incorrect diagnosis. Therefore, out-of-office BP measurements are often needed. Several studies have demonstrated the value of ambulatory BP (ABP) monitoring in pediatric hypertension, whereas home BP (HBP) monitoring has only recently been evaluated. METHODS: A systematic review of the evidence on HBP monitoring in children and adolescents has been performed (Medline/PubMed, Embase and Cochrane Library). RESULTS: A total of 27 studies (19 original study reports, two surveys, three guidelines documents, two reviews and one letter) were identified. These data suggest that by using electronic arm devices, reliable HBP readings are obtained. Unfortunately, very few electronic devices have been successfully validated in children. The reproducibility of HBP in children appears to be superior to office and similar to ABP measurements. Three-day-HBP monitoring with duplicate morning and evening measurements is the minimum schedule required, yet 6-7-day monitoring is recommended. HBP in children and adolescents is lower than daytime ABP, whereas no such difference exists in the adults. A school-based study in 778 children and adolescents provided the first HBP normalcy data. HBP has similar diagnostic value in children as in the adults and appears to be a reliable alternative to ABP monitoring in the detection of white-coat hypertension. CONCLUSION: HBP monitoring appears to have considerable potential in pediatric hypertension. More research is needed on the clinical application of this method in children and adolescents. PMID- 19542895 TI - Excessive sympathetic activation in heart failure with chronic renal failure: role of chemoreflex activation. AB - OBJECTIVE: Sympathetic activation contributes both to the initiation and progression of heart failure. The role of chronic renal failure (CRF) in determining sympathetic overactivity in chronic heart failure (CHF) patients is unknown. We tested the hypothesis that in CHF patients, CRF could lead to increase sympathetic activity through tonic activation of excitatory chemoreceptor afferents. METHODS: We conducted a double-blind, randomized, vehicle-controlled study to examine the effect of chemoreflex deactivation on muscle sympathetic nerve activity in CHF patients with or without CRF. We compared effect of breathing 100% oxygen for 15 min in 15 stable CHF patients with CRF and 15 control CHF patients matched for age, sex, blood pressure and BMI. RESULTS: The baseline muscle sympathetic nerve activity was significantly elevated in CHF patients with CRF as compared with simple CHF patients (61 +/- 3 versus 42 +/- 4 bursts/min; P < 0.01). Administration of 100% oxygen led to a significant decrease in muscle sympathetic nerve activity in CHF patients with CRF (from 61 +/- 3 to 55 +/- 4 bursts/min; P < 0.05). By contrast, neither 100% oxygen nor room air changed muscle sympathetic nerve activity or hemodynamics in patients with solely CHF. CONCLUSION: Tonic activation of excitatory chemoreflex afferents contributes to increased efferent sympathetic activity to muscle circulation and to blood pressure control in CHF patients with CRF. These findings may have important implications for understanding how CRF contributes to the progression of CHF and increases morbidity and mortality in CHF patients. PMID- 19542896 TI - Accuracy of colour duplex sonography for the diagnosis of renal artery stenosis. AB - OBJECTIVE: To examine the diagnostic value of novel velocimetric colour duplex sonography indices in the screening of renal artery stenosis (RAS). METHODS: We performed a retrospective analysis of all consecutively studied patients at our centre with suspected RAS, and a colour duplex sonography carried out at less than 4 months (mean 34 days) before renal angiography during a 6-year period (2002-2007). A significant RAS was defined as an at least 60% stenosis on angiography or a transstenotic mean arterial pressure gradient of at least 10 mmHg or both. RESULTS: In a total of 169 patients, 111 stenotic and 206 nonstenotic kidneys were examined. The sensitivity and specificity for acceleration of blood flow in early systole (ACCmax) were 85 and 75%, respectively, and for the acceleration index (ACCmax/peak systolic velocity, AImax) 83 and 79%, respectively. Corresponding values in patients with estimated glomerular filtration rate of less than 30 ml/min/1.73 m2 were 90 and 73% (for ACCmax) and 74 and 88% (for AImax). In addition, the transstenotic mean arterial pressure gradient showed a significant, though weak, negative correlation to ACCmax (r = -0.26, P = 0.02) and AImax (r = -0.29, P = 0.01) in stenotic kidneys. CONCLUSION: ACCmax and AImax provide similar, good diagnostic accuracy in the detection of a haemodynamically significant RAS, even in patients with markedly reduced glomerular filtration rate. Presumably, the lack of superiority of the novel index AImax could be explained by a highly homogenous methodological approach in the present single-centre study. PMID- 19542897 TI - The idiopathic environmental intolerance symptom inventory: development, evaluation, and application. AB - OBJECTIVE: To develop, evaluate, and apply a questionnaire-based instrument for investigation of specific symptoms in idiopathic environmental intolerance (IEI), called the Idiopathic Environmental Intolerance Symptom Inventory (IEISI). METHODS: Participants with IEI to chemicals responded to 82 candidate symptoms and to three subscales of the Quick Environmental Exposure and Sensitivity Inventory (QEESI) at a test (n = 207) and retest (n = 193) occasion. RESULTS: The 27 most commonly reported symptoms were selected and grouped into five symptom categories. Internal consistency, test-retest reliability, and concurrent validity were found to be satisfying. Cluster analysis identified two subgroups of IEI to chemicals. CONCLUSIONS: The results provide support for the IEISI being a reliable, valid, and fast tool for the study of specific symptom prevalence in IEI and encourage further study of subgroups. PMID- 19542898 TI - A review and analysis of the clinical and cost-effectiveness studies of comprehensive health promotion and disease management programs at the worksite: update VII 2004-2008. AB - OBJECTIVE: To conduct the seventh periodic review and analysis of the clinical and cost-effectiveness research conducted in worksite/corporate environments between 2004 and 2008. METHODS: A literature search of US-based research was conducted using a multistage process including MEDLINE, ADI, EDGAR, CARL, Inform, Lexis-Nexis, as well as direct inquiries to key researchers in this area of expertise. RESULTS: From 2004 to 2008, there were 16 new studies that met the Inclusion/Exclusion criteria. These studies are critiqued and entered into a data table consisting of 13 variables and citations of the 16 new studies. CONCLUSIONS: Clinical and cost-effectiveness research in the worksite/corporate environment continues to evolve although there was only one randomized clinical trial conducted during the most recent interval. There are innovations in pilot studies, quasi-experimental methodologies, and econometric modeling as indicative of future trends. PMID- 19542899 TI - Second-line combination chemotherapy with docetaxel and nedaplatin for Cisplatin pretreated refractory metastatic/recurrent esophageal squamous cell carcinoma. AB - BACKGROUND: There is an urgent need for an effective second-line chemotherapy regimen after failure of the standard cisplatin and 5-fluorouracil therapy. PATIENTS AND METHODS: This study investigated the efficacy and toxicity of the combination of docetaxel (30 mg/m2) during a 1-hour infusion, followed by nedaplatin (50 mg/m2) during a 2-hour infusion (both drugs were administered on day 1 as an outpatient regimen and repeated every 2 weeks) as second-line chemotherapy for patients with cisplatin-pretreated refractory metastatic/recurrent esophageal squamous cell carcinoma after surgery. RESULTS: Forty-six of the 48 patients (95.8%) were assessable for response. Partial response was confirmed in 13 of 48 cases yielding a response rate of 27.1% (95% confidence interval [CI], 14.5-39.7%). The median overall time to progression and overall survival was 3.1 months (95% CI, 2.3-3.9 months) and 5.9 months (95% CI, 3.9-7.8 months), respectively. The estimate of overall survival at 12 months was 16.7% (95% CI, 6.1-27.2%). Grade 3 anemia leucopenia, grade 4 anemia leucopenia and neutropenia were detected in only 4 (8.7%), 8 (17.4%), and 9 patients (19.6%), respectively. CONCLUSIONS: The combination chemotherapy of docetaxel and nedaplatin in the outpatient setting is well tolerated and useful as second-line chemotherapy for cisplatin-pretreated refractory metastatic/recurrent esophageal squamous cell carcinoma. PMID- 19542900 TI - Drosophila, a golden bug, for the dissection of the genetic basis of tolerance and susceptibility to hypoxia. AB - We have previously discovered that the adult Drosophila melanogaster is tolerant to a low O2 environment, withstanding hours of total O2 deprivation without showing any evidence of cell injury. Subsequently, our laboratory embarked on the study of hypoxia tolerance using a mutagenesis and overexpression screens to begin to investigate loss-of-function or gain-of-function phenotypes. Both have given us promising results and, in this article, we detail some of the interesting results. Furthermore, several years ago, we have also started an experimental "Darwinian" selection to generate a fly strain that can perpetuate through all of its life cycle stages in hypoxic environments. Through microarrays and bioinformatic analyses, we have obtained genes (e.g. Notch pathway genes) that play an important role in hypoxia resistance. In addition, we also detail a proof of principle that Drosophila genes that are beneficial in fly resistance to hypoxia can also be as well in mammalian cells. We believe that the mechanisms that we are uncovering in Drosophila will allow us to gain insight regarding susceptibility and tolerance to low O2 and will therefore pave the way to develop better therapies for ailments that afflict humans as a consequence of low O2 delivery or low blood O2 levels. PMID- 19542902 TI - The -174GG interleukin-6 genotype is protective from retinopathy and nephropathy in juvenile onset type 1 diabetes mellitus. AB - The aim of our study was to determine an association between the -174G>C IL-6 polymorphism (rs1800795) and occurrence of retinopathy and nephropathy in type 1 diabetes mellitus (T1DM) patients. Two hundred ten children/adolescents with long standing T1DM (16.5 +/- 3.8 y; with diabetes duration of 8.4 +/- 3.0 y) were enrolled into the study. A group of 170 healthy young (16.9 +/- 5.2 y) sex matched volunteers was qualified as the control. The IL-6 polymorphism was genotyped with the PCR-RFLP method. Serum and urine IL-6 concentrations were measured by the ultra-sensitive ELISA tests. The -174GG genotype was under represented in the diabetic patients compared with the controls. Patients with this genotype were free from nephropathy and retinopathy. The group of -174GG carriers was characterized by the highest urine IL-6 concentrations in relation to other genotypes. In the multivariate logistic regression analysis adjusted for age, duration of the disease, age of disease onset, HbA1c, and albumin excretion rate, the -174GG genotype was the only independent variable that significantly decreased the risk of jointly analyzed retinopathy and nephropathy [OR = 0.65; 95% CI = 0.52-0.82; p = 0.0003]. We propose that the -174GG patients are protected from late diabetic complications by different IL-6 dependent mechanisms. PMID- 19542901 TI - Early treatment with alglucosidase alpha prolongs long-term survival of infants with Pompe disease. AB - In a previous 52-wk trial, treatment with alglucosidase alpha markedly improved cardiomyopathy, ventilatory function, and overall survival among 18 children <7 mo old with infantile-onset Pompe disease. Sixteen of the 18 patients enrolled in an extension study, where they continued to receive alglucosidase alpha at either 20 mg/kg biweekly (n = 8) or 40 mg/kg biweekly (n = 8), for up to a total of 3 y. These children continued to exhibit the benefits of alglucosidase alpha at the age of 36 mo. Cox regression analyses showed that over the entire study period, alglucosidase alpha treatment reduced the risk of death by 95%, reduced the risk of invasive ventilation or death by 91%, and reduced the risk of any type of ventilation or death by 87%, compared with an untreated historical control group. Cardiomyopathy continued to improve and 11 patients learned and sustained substantial motor skills. No significant differences in either safety or efficacy parameters were observed between the 20 and 40 mg/kg biweekly doses. Overall, long-term alglucosidase alpha treatment markedly extended survival as well as ventilation-free survival and improved cardiomyopathy. PMID- 19542903 TI - A critical role for the IL-1 receptor in lung injury induced in neonatal rats by 60% O2. AB - IL-1 beta, a proinflammatory cytokine, may contribute to the development of the chronic neonatal lung injury, bronchopulmonary dysplasia. Chronic neonatal lung injury was induced in rats, by exposure to 60% O2 for 14 d from birth, to determine whether pulmonary IL-1 expression was up-regulated and, if so, whether a daily s.c. IL-1 receptor antagonist injections would be protective. Exposure to 60% O2 for 14 d caused pulmonary neutrophil and macrophage influx, increased tissue fraction and tyrosine nitration, reduced VEGF-A and angiopoietin-1 expression, and reduced small vessel (20-65 microm) and alveolar numbers. Lung IL 1 alpha and -1 beta contents were increased after a 4-d exposure to 60% O2. IL-1 receptor antagonist treatment attenuated the 60% O2-dependent neutrophil influx, the increased tissue fraction, and the reduced alveolar number. Treatment did not restore VEGF-A or angiopoietin-1 expression and only partially attenuated the reduced vessel number in 60% O2-exposed pups. It also caused a paradoxical increase in macrophage influx and a reduction in small vessels in air-exposed pups. We conclude that antagonism of IL-1-mediated effects can, in major part, protect against lung injury in a rat model of 60% O2-induced chronic neonatal lung injury. PMID- 19542904 TI - Elevated free thyroxine levels detected by a neonatal screening system. AB - In Sapporo city of Japan, neonatal screening for congenital hypothyroidism has used the measurement of free thyroxine (T4) and thyroid-stimulating hormone (TSH) in the filter-paper blood spot. This system has enabled us to identify hyperthyroxinemic diseases. Filter papers were collected from neonatal infants born at 4-6 d of age and neonates who showed elevated free T4 (>4.0 ng/dL, 4 SD above the mean) were studied. Between January 2000 and December 2006, 83,232 newborns were screened. Eleven infants demonstrated persistent hyperthyroxinemia. One patient with slightly elevated free T4 and normal TSH was diagnosed as having familial dysalbuminemic hyperthyroxinemia (FDH). The other two patients with elevated free T4 without suppressed TSH were considered as having resistance of thyroid hormone (RTH), and analysis of thyroid hormone receptor (TR) beta gene confirmed the diagnosis. The remaining eight patients were diagnosed as having neonatal Graves' disease (NGD). Seven of eight pregnant women were treated with antithyroid drug and thus only one unrecognized NGD during pregnancy was detected by screening. Our screening system enables for early awareness of RTH and FDH. Regarding Graves' disease, the benefit of elevated free T4 screening is small, because most pregnant women with Graves' disease were managed. PMID- 19542905 TI - Effect of sustained inflation length on establishing functional residual capacity at birth in ventilated premature rabbits. AB - The effect of inflation length on lung aeration pattern, tidal volumes, and functional residual capacity (FRC) immediately after birth was investigated. Preterm rabbits (28 d), randomized into four groups, received a 1-, 5-, 10-, or 20-s inflation (SI) followed by ventilation with 5 cm H2O end-expiratory pressure. Gas volumes were measured by plethysmography and uniformity of lung aeration by phase contrast x-ray imaging for 7 min. The first inspiratory volume significantly (p < 0.001) increased with inflation duration from a median (IQR) of 0.2 (0.1-3.1) mL/kg for 1-s inflation to 23.4 (19.3-30.4) mL/kg for 20-s SI. The lung was uniformly aerated, and the FRC and tidal volume fully recruited after 20-s SI. A 10-s SI caused a higher FRC (p < 0.05) at 7 min, and a 20-s SI caused a higher FRC (p < 0.05) at 20 s and 7 min than a 1- or 5-s SI. The mean (SD) time for 90% of the lung to aerate was 14.0 (4.1) s using 35 cm H2O peak inflation pressure. In these rabbits, 10- and 20-s SI increased the inspiratory volume and produced a greater FRC, and a 20-s SI uniformly aerated the lung before ventilation started. PMID- 19542906 TI - Impaired voltage gated potassium channel responses in a fetal lamb model of persistent pulmonary hypertension of the newborn. AB - We investigated the hypothesis that oxidative stress in persistent pulmonary hypertension of the newborn (PPHN) impairs voltage gated potassium (Kv) channel function. We induced PPHN in fetal lambs by prenatal ligation of ductus arteriosus; controls had sham ligation. We studied changes in the tone of pulmonary artery (PA) rings and Kv channel current of freshly isolated PA smooth muscle cells (PASMC) using standard techniques. 4-Aminopyridine (4-AP), a Kv channel antagonist, induced dose-dependent constriction of control PA rings; this response was attenuated in PPHN pulmonary arteries. Exogenous superoxide and peroxynitrite inhibited the response to 4-AP in control rings. Tiron, a superoxide scavenger, improved the response to 4-AP in PPHN rings. 4-AP inhibited the NOS-independent relaxation response to ATP in control PA rings. Relaxation response to ATP was blunted in PPHN rings and was improved by NOS antagonist, N nitro-L-arginine methyl ester (L-NAME). 4-AP attenuated this response in L-NAME treated PPHN rings. Exogenous superoxide suppressed 4-AP sensitive Kv current in control PASMC. Kv channel current was attenuated in cells from PPHN lambs and was restored by tiron. Oxidative stress impairs Kv channel function in PPHN. Superoxide scavengers may improve pulmonary vasodilation in PPHN in part by restoring Kv channel function. PMID- 19542907 TI - Antenatal corticosteroids increase fetal, but not postnatal, pulmonary blood flow in sheep. AB - The lungs of very preterm infants have immature airways and gas exchange structures and are usually surfactant deficient. Antenatal corticosteroids are commonly used to enhance fetal lung maturation in preterm infants, but little is known of their effects on pulmonary blood flow (PBF) before and immediately after birth. Our aim was to determine the effects of antenatal betamethasone on PBF before birth and during the postnatal transition in very preterm lambs. Antenatal betamethasone treatment significantly increased mean fetal PBF from 20.2 +/- 5.1 to 84.3 +/- 18.3 mL/min at 30 h after administration; the PBF waveform was also significantly altered. Mean diastolic PBF increased from -38.5 +/- 4.9 pretreatment to -10.2 +/- 11.0 mL/min at approximately 36 h after the initial betamethasone dose (negative values indicate retrograde flow away from the lungs). Within 10 min after delivery, PBF was similar in control and betamethasone-treated lambs. These data demonstrate that antenatal betamethasone significantly increases fetal PBF and alters the PBF waveform but has little effect on postnatal PBF. PMID- 19542908 TI - The relationship of insulin resistance with SNP 276G>T at adiponectin gene and plasma long-chain polyunsaturated fatty acids in obese children. AB - This study examined the association of insulin resistance with single-nucleotide polymorphism (SNP) 276G>T at adiponectin gene and the plasma long-chain polyunsaturated fatty acids (LCPUFAs) profile in obese children. One hundred thirty-one normolipidaemic obese children aged 8-13 y (53 girls and 68 boys) entered the study. The prevalence of T allele carriers at SNP276 was 48.8%. Mean [SD] values of fasting insulin and homeostasis model assessment-insulin resistance (HOMA-IR) index in noncarriers versus carriers of T allele were 12.4 [6.4] versus 20.6 [6.3] muU/mL (p = 0.039) and 2.6 [1.4] versus 4.5 [1.7] (p = 0.032). Mean [SD] values of plasma C18:3n - 3, C20:5n - 3/C20:4n - 6, and n - 6/n - 3 LCPUFA in phospholipids in noncarriers versus carriers of T allele were 0.10 [0.04] versus 0.08 [0.03] % (p = 0.013), 0.04 [0.01] versus 0.03 [0.01] % (p = 0.045), and 4.4 [0.7] versus 4.9 [0.9] % (p = 0.005), respectively. Insulin resistance was independently associated with SNP 276G>T (p = 0.002) and n - 6/n - 3 LCPUFA (p = 0.042) in plasma phospholipids, and interaction was found between SNP 276G>T and n - 6/n - 3 LCPUFA (p = 0.046). These findings suggest that obese children carriers of the SNP 276G>T may be at increased risk of metabolic complications compared with noncarriers, possibly due in part to a different plasma phospholipids profile. PMID- 19542909 TI - Sex differences in a hypoxia model of preterm brain damage. AB - Male sex is a well-established risk factor for poor neurodevelopmental outcome after premature birth. The mechanisms behind this sex-related difference are unknown. The damage associated with prematurity can be mimicked in rodents by prolonged exposure to sublethal postnatal hypoxia. This chronic hypoxia leads to anatomical changes in mice that strongly resemble the loss of volume, decreased myelination, and ventriculomegaly seen in preterm newborns. However, no sex differences have been previously noted in this rodent model. We hypothesized that sex comparisons in hypoxic mice would show sex-related differences in brain volume and white matter loss in response to the same degree of hypoxic insult. Mice were placed in chronic sublethal hypoxia from postnatal day 3-11. Cortical, hippocampal, and cerebellar volumes and myelination indices were measured. We found that the male hippocampus, normally larger than the female, undergoes a greater volume loss compared with females (p < 0.05). Myelination, generally greater in males, was significantly disrupted by hypoxia in neonatal male forebrain. These results support the use of this rodent model to investigate the basis of sex-related susceptibility to brain damage and develop new sex-based neuroprotective strategies. PMID- 19542910 TI - Siglec-9 and SHP-1 are differentially expressed in neonatal and adult neutrophils. AB - Neonatal PMN (polymorphonuclear neutrophils) exhibit altered inflammatory responsiveness and greater longevity compared with adult PMN; however, the involved mechanisms are incompletely defined. Receptors containing immunoreceptor tyrosine-based inhibitory motif (ITIM) domains promote apoptosis by activating inhibitory phosphatases, such as Src homology domain 2-containing tyrosine phosphatase-1 (SHP-1), that block survival signals. Sialic acid-binding immunoglobulin-like lectin (Siglec)-9, an immune inhibitory receptor with an ITIM domain, has been shown to induce cell death in adult PMN in association with SHP 1. To test our hypothesis that neonatal PMN inflammatory function may be modulated by unique Siglec-9 and SHP-1 interactions, we compared expression of these proteins in adult and neonatal PMN. Neonatal PMN exhibited diminished cellular expression of Siglec-9, which was phosphorylated in the basal state. Granulocyte-macrophage colony-stimulating factor (GM-CSF) treatment decreased Siglec-9 phosphorylation levels in neonatal PMN but promoted its phosphorylation in adult PMN, observations associated with altered survival signaling. Although SHP-1 expression was also diminished in neonatal PMN, GM-CSF treatment had minimal effect on phosphorylation status. Further analysis revealed that Siglec-9 and SHP-1 physically interact, as has been observed in other immune cells. Our data suggest that age-specific interactions between Siglec-9 and SHP-1 may influence the altered inflammatory responsiveness and longevity of neonatal PMN. PMID- 19542912 TI - Immunohistochemical expression of S100A6 in cellular neurothekeoma: clinicopathologic and immunohistochemical analysis of 31 cases. AB - Neurothekeoma is a term introduced by Gallager and Helwig describing a superficial tumor of purported nerve sheath derivation, with cellular and myxoid types. Recently, it has been suggested that the cellular type does not have nerve sheath differentiation. This subtype represents an uncommon neoplasm and sometimes can be problematic to diagnose because it can be easily mistaken for melanoma. We studied the immunohistochemical features of 31 cases of cellular neurothekeomas to evaluate their immunoprofile. Immunohistochemical studies were performed in all 31 cases with formalin-fixed paraffin-embedded tissue sections with antibodies against S100 protein, S100A6, and melanoma antigen recognized by T-cells (MART-1). In addition, 8 cases were evaluated for HMB-45 antigen, keratin (with a pankeratin cocktail), epithelial membrane (EMA), and smooth muscle antigen (SMA). The lesions were from 8 men and 23 women aged 6-64 years (mean 35 years). Four tumors were located on the nose; 4 scalp; 4 finger; 3 thigh; 2 shoulder; 2 wrist; 2 hand; and 1 each on pelvis, cheek, toe, chest, eyebrow, forearm, penis, axilla, mouth, and leg. All tumors were positive for S100A6 (100%) and negative for cytokeratin, HMB-45 antigen, MART-1, and EMA (100%); 29 cases were negative for S100 protein (93.5%; the 2 positive cases had only scattered cells labeled), and only 2 cases were focally positive for SMA (7.5%). Therefore, the combination of strong immunoreactivity for S100A6, in nested dermal spindle cell proliferations, and lack of S100 protein or keratin, supports a diagnosis of cellular neurothekeoma. PMID- 19542911 TI - Physiological partial aldosterone resistance in human newborns. AB - In the neonatal period, the human kidney is characterized by an impaired ability to regulate water and sodium homeostasis, resembling partial aldosterone resistance. The aim of our study was to assess this hormonal insensitivity in newborn infants and to determine its relationship with neonatal sodium handling. We conducted a prospective study in 48 healthy newborns and their mothers. Aldosterone, renin, and electrolyte concentrations were measured in umbilical cords and in maternal plasma. Urinary aldosterone concentrations and sodium excretion were determined at urination within 24 h after birth. A significant difference was observed between aldosterone and renin levels in newborn infants compared with their mothers (817 +/- 73 versus 575 +/- 55 pg/mL and 79 +/- 10 versus 15 +/- 2 pg/mL, respectively, p < 0.001). This hyperactivation of the renin-angiotensin-aldosterone system was associated with hyponatremia and hyperkalemia in the newborn infants, and high urinary sodium loss, consistent with a partial aldosterone resistance at birth. Unlike plasma aldosterone, urinary aldosterone concentration was found highly correlated with plasma potassium concentrations, thus representing the best index for accurate evaluation of mineralocorticoid sensitivity. Our study represents a comprehensive characterization of the renin-aldosterone axis in newborn infants and provides evidence for physiologic partial aldosterone resistance in the neonatal period. PMID- 19542913 TI - Low-fat and fat-free pleomorphic lipomas: a diagnostic challenge. AB - Pleomorphic lipomas are benign tumors that most commonly present as subcutaneous masses in the head and neck, shoulder, or back region of middle-aged to elderly men. They are related to spindle cell lipomas based on shared cytogenetic aberrations and histologic features. When little or no fat is present, the diagnosis can be challenging. A review of 38 pleomorphic lipomas seen in consultation revealed 7 cases in which fat was present in reduced (<5%) amounts (n = 5) or absent (n = 2). Six of 7 cases were from men with a mean age of 59 years. Excluding 1 case where the site was not specified, they all presented as solitary well-circumscribed subcutaneous masses in the head and neck (n = 3) or shoulder (n = 2) region. The seventh case was an intradermal tumor from the nose of a 48-year-old woman. All displayed pleomorphic and multinucleated floret cells interspersed among bland spindle cells and ropey collagen. They were diffusely immunoreactive for CD34. Referring diagnoses, when provided, included myxofibrosarcoma, giant cell fibroblastoma, and granulomatous rosacea for the tumor from the nose; none considered pleomorphic lipomas. When fat is absent or present in reduced amounts, clinical context and identification of classic nonlipogenic components are essential for the diagnosis of pleomorphic lipomas. PMID- 19542914 TI - Differential expression of the antioxidant repair enzyme methionine sulfoxide reductase (MSRA and MSRB) in human skin. AB - Recently, the antioxidant repair enzymes methionine-S-sulfoxide reductase A (MSRA) and methionine-R-sulfoxide reductase B (MSRB) were described in human epidermal keratinocytes and melanocytes. Methionine sulfoxide reductases (MSRs) are thought to protect against reactive oxygen species-induced oxidative damage in many organs, including the most environmentally exposed organ, human skin. We sought to examine the expression and distribution of this enzyme family (MSRA, MSRB1, MSRB2, and MSRB3) within the various compartments of healthy and diseased human skin. Expression was assessed using polyclonal MSR antibodies and immunohistochemical staining of human skin biopsies from various anatomical sites. Remarkably, MSRA expression was not only found in the epidermis as previously described but also in hair follicles and eccrine glands and was most pronounced in sebaceous glands. Furthermore, MSRB2 expression was found in melanocytes while MSRB1 and MSRB3 were both expressed within vascular endothelial cells. In conclusion, MSR enzymes are differentially expressed in human skin. Thus, modulation of MSR repair antioxidants may have implications for cutaneous aging and carcinogenesis. PMID- 19542915 TI - The utility of PU.1 as an immunohistochemical marker for histiocytic and dendritic lesions of the skin. AB - PU.1 is a transcription factor restricted to the hematopoietic system. It is expressed in myeloid lineage and B lymphocytes but is absent in mature T cells and nonhematopoietic cells. Among myeloid lineage-derived cells, PU.1 is overexpressed in monocytes, histiocytes, and dendritic cells. We evaluated PU.1 expression in 78 cases of primary skin neoplasms, including 9 reticulohistiocytomas, 9 Langerhans cell histiocytoses, 7 juvenile xanthogranulomas, 9 fibrous papules, 8 dermatofibromas, 12 dermatofibrosarcoma protuberans, 9 Spitz nevi, and 15 malignant melanomas. Strong nuclear staining for PU.1 was seen in all cases of histiocyte and dendritic cell origin, including 9/9 reticulohistiocytomas, 9/9 Langerhans cell histiocytoses, and 7/7 juvenile xanthogranulomas. No staining for PU.1 was seen in any studied cases of fibrous papules, dermatofibrosarcoma protuberans, dermatofibromas, Spitz nevi, or malignant melanomas. This study indicates that PU.1 is a valuable immunohistochemical marker for identifying cutaneous histiocyte- and dendritic cell-derived lesions. PU.1 staining is easily interpreted due to the sharp nuclear staining as compared with the irregular and often variable cytoplasmic staining seen with CD68. PMID- 19542916 TI - Eruptive syringoma of the penis. A report of 2 cases and a review of the literature. AB - Syringoma exclusively located to the penis is an extraordinary rare dermatological condition, and only 6 patients with syringoma exclusively located to the penis have been published previously. Light microscopical examination of routine hematoxylin and eosin-stained sections from punch biopsies with adequate depth from 2 patients was performed. Classical syringoma was diagnosed. The lesions occurred in an eruptive fashion in both patients on the dorsal and lateral aspects of the penis shaft. There was a variation in clinical appearances ranging from yellow-white papules, a few millimeter in size to, in one of the patients, formation of several slightly elevated plaques, 10 mm in size, with a brownish-red color. We describe clinicopathological features of 2 patients with syringoma exclusively located to the penis. These are compared with all the previously published cases. Clinical and histopathological differential diagnoses are discussed, and a literature review is presented. PMID- 19542917 TI - Ingrown toenail: histopathologic and immunohistochemical study. AB - Although ingrown toenail (IT) is a common problem, the morphologic alterations presented in this condition have rarely been presented in literature. We have studied 14 cases from 12 different patients and studied the biopsies of all of them, not only with routine techniques but also with histochemical stains (Masson trichrome, periodic acid-Schiff, and Gram) and with immunohistochemical techniques mindbomb homolog-1 (MIB-1), p63, D2-40, and AE1-AE3). The patients were clinically classified into 4 stages (1, 2a, 2b, and 3). The main changes evidenced were ingrowing of the squamous epithelium (either from the matrix or mostly from the hyponichium) and chronic plasma-cell rich inflammation with prominent vascularization. The intensity of these changes varied depending on the stage: whereas they were mild in stages 1 and 2a, they were prominent in stages 2b and 3. In advanced stages, the epithelial nests showed certain dyskeratosis. The expression of MIB-1 was, nevertheless, only basal and parabasal. p63 was intensively expressed by the epithelial nests. We therefore conclude that (a) IT shows epithelial ingrowing with accompanying granulation tissue; (b) according to the clinical stage of the disease, this pattern can be more or less complex; (c) in advanced stages of IT, the pattern can mimic a well-differentiated squamous cell carcinoma; (d) MIB-1 is not helpful in distinguishing IT from squamous cell carcinoma; (e) the high expression of p63 by the ingrowing epithelium should not be the source of a misdiagnosis of malignancy. PMID- 19542918 TI - Requirement of DHCR24 for postnatal development of epidermis and hair follicles in mice. AB - Desmosterolosis is an autosomal recessive disorder with severe developmental anomalies due to mutations in the DHCR24 gene, encoding an enzyme to convert desmosterol to cholesterol. We reported that DHCR24 [knockout (KO)] mice were born with wrinkleless taut skin and with impaired development of epidermis. In this study, we investigated the postnatal development of epidermis and hair follicle in the skin of KO mice grafted to the nude mice. Skin graft was required since the KO mice die within few hours after birth. Forty days after the skin graft, epidermis from the KO mice revealed the characteristic phenotype observed at birth. Furthermore, the number of hair follicles in the skin graft from KO mice to the nude mice was significantly less and development was delayed than that from control. These findings implicate that DHCR24 plays important roles for normal development of epidermis and hair follicle even in postnatal life. PMID- 19542919 TI - Neurofibroma with clear cell change. AB - Cutaneous clear cell proliferations and degenerative change have been seen in a variety of entities including nevi, dermatofibromas, fibrous papules, atypical fibroxanthomas, basal cell carcinomas, and squamous cell carcinomas, to name a few. However, there have been no reports of clear cells within neurofibromas. We received a biopsy and excision from a 61-year-old man with a papule on his right lateral clavicle. The initial biopsy showed a proliferation of clear cells that stained positive with S-100 and focally with CD68. A clear cell granular cell tumor was favored. Subsequent excision showed the same population of clear cells as seen on the initial biopsy. Interestingly, a neurofibroma was also present immediately beneath the clear cells with areas of transition. A p75 stain highlighted both populations of cells. This is the first case of neurofibroma with clear cells reported in the literature. We postulate that the clear cells are due to degenerative change. PMID- 19542920 TI - An unusual squamo-melanocytic tumor of uncertain biologic behavior: a variant of melanoma? AB - The presence of two intermingled malignant components within the same cutaneous tumor is rare. Herein, we report a case of an exceedingly rare squamo-melanocytic tumor, favored to be a melanoma variant. Although the biologic behavior and prognostic significance of this tumor remain to be established, this unique case reaffirms the potential morphologic diversity of melanoma. We review the literature reporting similar neoplasms and discuss the potential histogenesis of this squamo-melanocytic tumor. PMID- 19542921 TI - Cutaneous focal mucinosis causing follicular induction of the epidermis. AB - Cutaneous focal mucinosis has been rarely reported in association with follicular induction of the epidermis. We present 2 cases of focal mucinosis with follicular induction and describe the histopathologic findings to create awareness of this association and to prevent confusion with other diagnoses such as dermatofibroma with follicular induction or superficial basal cell carcinoma. PMID- 19542922 TI - A vulvar fibroepithelial stromal polyp appearing in infancy. AB - We present here a first case of vulvar fibroepithelial stromal polyp (vFSP) appearing in infancy, including previously undescribed papule as an early form of vFSP. A 7-month-old girl presented with a small (adzuki-sized), soft, erythematous papule in the right labium majus. The size of the lesion increased up to 2.3 cm in the following 7 months and exhibited cerebriform or frond-like configuration. The surgically removed polyp was characterized by the proliferation of bland spindle cells with tapered cytoplasm, and the diagnosis of vFSP was established. Twenty-one months after the first surgery, a recurrent polyp 2.0 cm in size was resected. No further recurrence has been observed in the 17 months since the second surgery. PMID- 19542923 TI - Synchronous presence of cellular angiofibroma and lipoma in vulvoinguinal region: a unique case report. AB - A 26-year-old woman presented with a 7-month history of a slowly growing right sided, well-circumscribed, subcutaneous, soft, painless, large vulvoinguinal lump that now measured up to 12 cm. Adjacent to but separate from this was a well circumscribed, soft, painless, mobile, large inguinal tumor 8 cm in diameter that had been present unchanged for 2 years. Both lesions were completely excised. Histologically, the first mass was consistent with cellular angiofibroma, whereas the second one had the typical features of lipoma. According to the available literature, this is the first case of synchronous presence of cellular angiofibroma and lipoma in one patient. PMID- 19542924 TI - Nodular amyloidosis: differentiation from colloid milium by electron microscopy. AB - Nodular amyloidosis is a primary cutaneous amyloidosis characterized by the deposition of amyloid L-type fibril proteins in the dermis. Clinical history and routine histology may not be sufficient to differentiate nodular amyloidosis from colloid milium. We present a case of a 45-year-old man with nodular amyloidosis, whose diagnosis was confirmed by the characteristic appearance of filaments on electron microscopy. PMID- 19542925 TI - Resolution of CD8+ lymphomatoid papulosis after surgical excision of the type AB thymoma. AB - Lymphomatoid papulosis (LyP) is a disease characterized by cyclic eruptions of papulonodular lesions, which undergo spontaneous healing. Lymphoid malignancies are present in 10%-15% of cases. The type AB thymoma is an epithelial neoplasm composed of both type A (lymphocyte-poor) and type B (lymphocyte-rich) areas. Chemokines are chemotactic cytokines, responsible for leukocyte motility and direct movement. Interactions between chemokines and their receptors have been correlated with homing of lymphoma cells to various tissues. We describe a patient whose type B LyP CD8 lesions completely resolved after surgical removal of type AB thymoma. The chemokine profile was similar in both tissues: thymus- and activation-regulated chemokine and CCR4 were focally positive in the thymoma. Thymus- and activation-regulated chemokine was positive in the epidermotropic cells and in the majority of the dermal lymphocytes in the LyP specimen, whereas CCR4 was focally positive in the dermal lymphocytes. Monokine induced by interferon-g (Mig) staining showed strong positivity in the dermal lymphocytes and in localized areas of the thymoma, but an immunostain for the Mig receptor (CXCR3) highlighted only a few scattered cells in both tissues (less than 3%). Both tissues were negative for regulated upon activation, normal T-cell expressed and secreted (RANTES) and CCR3. In summary, we report the association of a CD30 negative CD8-predominant LyP and a type AB thymoma, with similar chemokine profiles. The rarity of both conditions and the precise regression of LyP after removal of the thymoma argue against a coincidental observation. We recommend that a search for thymoma be included in the workup of LyP. Further chemokine profiling in other cases of LyP may assist in understanding their role in this disease. PMID- 19542926 TI - Trichogerminoma. AB - A case of distinctive benign follicular neoplasm previously reported under the designation of trichogerminoma is described. A 45-year-old man presented with an asymptomatic nodule on the scalp since 3 years. Histologically, the lesion corresponded to a well-organized, symmetrical dermal nodule made up of basophilic lobules included in a fibrocytic stroma. The lesion had the characteristics of hair germ tumors; however, most lobules depicted a distinctive pattern of rounded nests of concentrically arranged clear cells. Small follicle bulb-like basophilic structures, foci of sebaceous differentiation, and areas of infundibulocystic, isthmic, and outer sheath keratinization were also seen. This neoplasm and the other tumors with hair germ differentiation such as trichoblastoma and panfolliculoma seem to represent the same spectrum of hair follicle neoplasms only distinguishable by their degree of differentiation. PMID- 19542927 TI - Cutaneous solitary neural hamartoma: report of an unusual case. AB - Cutaneous hamartomas are tumor-like proliferations of tissue indigenous to the organ but arranged abnormally. There are examples in the literature of cutaneous hamartomas composed of a variety of different components. To our knowledge, there is no previous report of such cutaneous solitary neural hamartoma. Our case occurred in a 51-year-old man with pain and paresthesia in the right shoulder associated with a nodule that was surgically removed. There was no history of trauma, other skin nodules, neurofibromatosis, or tuberous sclerosis. Histologically, there was an unencapsulated nodule, composed of mature nerve bundles noted abnormally high within the papillary dermis, extending to the reticular dermis with periadnexal distribution. Immunohistochemically, the nerve bundles were positive for S-100, including the smaller nerve twigs, and the perineurium was highlighted by epithelial membrane antigen, reminiscent of normal peripheral nerves. Although, neural components including mature nerve bundles have been described in various cutaneous hamartomas, this represents a peculiar case of a cutaneous mature peripheral nerve hamartoma. Whether this is related to other entities of cutaneous hamartomas (ie, neurofollicular hamartoma, folliculosebaceous cystic hamartoma) is not yet apparent, although it is probably a unique entity. PMID- 19542928 TI - Cutaneous rosai-dorfman disease and morphea: coincidence or association? AB - Rosai-Dorfman disease or sinus histiocytosis with massive lymphadenopathy is a rare disease. A third of patients with this disease have extranodal involvement affecting the skin. Of these individuals, only around 3% will have purely cutaneous Rosai-Dorfman disease, which is limited to skin manifestations without systemic involvement. Cutaneous (localized) scleroderma or morphea, on the other hand, is a more common disease that most often affects women of all ages. Both conditions have unknown etiologies. Presented here is a case of a 60-year-old white woman with cutaneous Rosai-Dorfman disease and coexisting morphea. Representative biopsies from both areas were performed: one showing a dermal S 100+ histiocytic infiltrate with emperipolesis and the other showing a deep perivascular and interstitial plasma cell infiltrate with dermal sclerosis and loss of perieccrine fat. A laboratory and radiologic workup revealed no evidence of systemic involvement by either entity. The diagnosis of coexisting cutaneous Rosai-Dorfman disease and morphea was established. To our knowledge, this is the first report of these 2 entities found simultaneously in 1 patient. PMID- 19542929 TI - Metastatic desmoplastic malignant melanoma associated with low-grade myofibroblastic sarcoma. AB - The authors report a 63-year-old man with a 2-year history of a primary desmoplastic malignant melanoma on the right heel with lymph node metastasis in the right groin, presented with a large painless mass in the right inguinal area. Microscopically, there was a dense spindle cell proliferation in the reticular dermis, subcutis, and subjacent soft tissue. The neoplastic cells were moderately pleomorphic and showed mitotic figures including atypical ones. Immunohistochemical studies showed a biphasic cellular composition, with one component being S-100 protein positive and alpha-smooth muscle actin negative and another component with a reverse phenotype, namely, alpha-smooth muscle actin positive and S-100 protein negative. Pleomorphic cells and atypical mitotic figures were evident in both components. Electron microscopy identified cells with peripheral bundles of thin cytoplasmic myofilaments with focal dense bodies, fibronexus junctions, abundant rough endoplasmic reticulum, micropinocytic vesicles, and strikingly folded nuclei. These cells had features of myofibroblasts. We conclude that the myofibroblastic proliferation in this case clearly exceeded just a stromal reaction and fulfilled the criteria of a low grade sarcoma. PMID- 19542930 TI - Squamomelanocytic tumor: an unusual and distinctive entity of uncertain biological potential. AB - We report a case of a squamomelanocytic tumor of the skin. Clinically, the lesion was felt to be a melanocytic or vascular tumor but histologically was characterized by epithelioid cells with focal squamous differentiation. Immunohistochemical staining showed that half of cells stained with MNF116 and a smaller proportion stained with S100 and Melan A. A third population did not stain with either set of antigens. The lesion has some similarities to a melanocytic matricoma but no evidence of matrical differentiation. The biological potential of this distinctive tumor is not known because so few have been reported. PMID- 19542932 TI - Skin metastasis from hepatocarcinoma. AB - Cutaneous metastases may be the first sign of an internal neoplasm; hepatocellular carcinoma is a very rare source of skin involvement and presents with a protean morphology: papules, nodules, and masses that may be by natural sources or by artificial mechanisms. We present a 41-year-old man with hepatocellular carcinoma and hepatic cirrhosis who developed metastatic disease, which in the skin showed as a pyogenic granuloma. We analyze 38 cases from the literature and present a practical review of the subject. PMID- 19542931 TI - Subcutaneous atypical fibrous histiocytoma. AB - Benign fibrous histiocytoma is one of the most frequent benign neoplasms mainly composed of a mixture of fibroblastic and histiocytic cells, especially found in the skin (dermatofibroma), particularly in the limbs. The diagnosis of cutaneous benign fibrous histiocytoma is generally easy; however, rare variants may be difficult to identify, and the diagnosis only confirmed after exhaustive histopathological examination. Thus, deep subcutaneous dermatofibroma may be difficult to distinguish from dermatofibrosarcoma protuberans and dermatofibroma with monster giant cells from malignant fibrous histiocytoma and atypical fibroxanthoma. We report a case of a 38-year-old woman with a painless swelling on the abdominal wall, which was totally excised and histopathologically diagnosed as subcutaneous atypical fibrous histiocytoma. The lesion was deeply located within the subcutaneous tissue and consisted of interlacing fascicles of predominant histiocyte-like spindle cells intermingled with pleomorphic giant cells with bizarre large nuclei (bilobed and multilobed) and prominent eosinophilic nucleoli. Only 1 mitotic figure was found in the whole lesion. Prominent hyaline collagen bundles surrounded by tumor cells were observed, predominantly at the periphery of the lesion. Immunohistochemical study showed positivity only for vimentin and factor XIIIa, whereas pan-keratins, actin, desmin, CD34, CD10, and S-100 protein were negative. Recognition of dermatofibroma is important, allowing sequential excision and optimal results. Definitive diagnosis, although especially difficult in our case, is established by characteristic histological and immunohistochemical criteria. To the best of our knowledge, we report the first case of subcutaneous fibrous histiocytoma with monster cells. PMID- 19542933 TI - A verrucous plaque of the eyebrow. AB - Phaeohyphomycosis is a rare dematiaceous fungal infection requiring surgical excision or appropriate systemic antifungal therapy. The objective of this study is to report a case of phaeohyphomycosis of the right eyebrow successfully treated with surgical excision. We performed an excision of a dermal plaque of phaeohyphomycosis using 4-mm margins on the right brow extending onto the right forehead. An island pedicle flap was used to close the defect. Histopathologic evaluation confirmed the presence of numerous dematiaceous fungal organisms fully consistent with phaeohyphomycosis, and the margins were verified to be free of fungal organisms. Culture of a central portion of the excised lesion confirmed that the causative organism was a saprophytic fungus. Adjuvant systemic antifungal therapy was offered to the patient, but because of financial resources, the patient did not desire any further treatment. The patient remains free of the organism and has healed well, without complications. Phaeohyphomycosis can be effectively treated with excision alone, and when located in a risky anatomic location, such as near the orbit, adjuvant treatment with systemic antifungals may be warranted to prevent recurrence. PMID- 19542934 TI - Lichen planus esophagitis: diagnostic implications. PMID- 19542935 TI - Diagnosis of cutaneous sarcomatoid B-cell lymphoma: some words of caution. PMID- 19542936 TI - Congenital plaque-like glomangioma of the scalp. PMID- 19542938 TI - Differences between multifibrin U and conventional Clauss fibrinogen assays: data from UK National External Quality Assessment Scheme surveys. PMID- 19542939 TI - ABO blood group does not increase the risk of thrombosis in Philadelphia-negative myeloproliferative disorders. PMID- 19542940 TI - Bleeding rates in elderly patients on vitamin K antagonist therapy for nonvalvular atrial fibrillation. PMID- 19542941 TI - Lymphoscintigraphy in angiomyomatous hamartomas and primary lower limb lymphedema. AB - PURPOSE: Angiomyomatous hamartoma (AH) of the lymph node is a rare vascular benign disease of unknown etiology with a predisposition for the lymph nodes of the inguinal area. Only 18 cases have been described up to now in the literature and the disorder was reported to be associated with lymphedema or swelling of the ipsilateral limb in 4 patients. However, scintigraphic investigation of the lymphatic system in these patients was reported in only 2 cases. MATERIAL AND METHODS: Five patients where the biopsy of inguinal nodes for suspected lymphadenitis led to the diagnosis of angiomyomatous hamartoma were investigated using lymphoscintigraphic techniques (1 girl aged 15; 1 boy aged 9 at the time of first biopsy and 11 at the time of the second one; and 3 men aged 30, 50, and 57). The operated limb was lymphedematous in 3 and 1 developed lymphedema after biopsy. The fifth patient developed a contralateral lymphedema after his second nodal biopsy. RESULTS: In all cases, lymphoscintigraphic investigation of the limbs showed extensive lymph node abnormalities on the operated side and in 4 cases on the opposite side. CONCLUSIONS: These observations support not only the hypothesis that lymphatic disturbance was involved in the pathogenesis of these tumors but also the proposition that lymphoscintigraphy should be performed in cases of inguinal lymphadenitis of unknown origin to diagnose the underlying situation of latent lymphedema. PMID- 19542942 TI - Radionuclide cisternography in detecting cerebrospinal fluid leak in spontaneous intracranial hypotension: a series of four case reports. AB - Spontaneous intracranial hypotension (SIH) is an infrequent clinical entity characterized by cerebrospinal fluid (CSF) hypovolemia due to a CSF leak. The cause of the leak in SIH, however, is largely unknown, though structural meningeal weakness and mechanical stress factors have been postulated. Patients with SIH typically present with postural headaches, and occasionally with other symptomology as well, such as nausea, emesis, neck stiffness, and photophobia. In this case series, we present 4 patients who underwent radionuclide cisternography (RNC) for suspected CSF leak. All patients underwent RNC and MR and/or CT for evaluation. We found that RNC accurately detected and localized a CSF abnormality in all 4 patients, with each patient experiencing symptomatic relief following directed epidural blood patch. PMID- 19542943 TI - Progressive osteoblastic bone metastases in breast cancer negative on FDG-PET. AB - Positron emission tomography using F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) is increasingly used in breast cancer. The new generation cameras integrate PET and CT within the same camera, allowing the simultaneous assessment of the structural and metabolic aspects of disease. There is presently a controversy on the clinical significance of osteoblastic bone metastases in breast cancer which are not detected on FDG-PET. It has been suggested that these radiologically dense lesions represent the result of successful treatment of initially osteolytic lesions. We report a case of a 65-year-old woman with a suspicion of recurrent breast cancer based on an increasing serum tumor marker. Serial PET/CT showed progressive blastic bone metastases on the CT without FDG uptake. These lesions were confirmed by bone single photon emission computed tomography. This case report shows: first, that progressive osteoblastic lesions can lack FDG-avidity, leading to a false-negative PET; and secondly, that bone scintigraphy should not be replaced by FDG-PET/CT for the detection of bone metastases in breast cancer. PMID- 19542944 TI - Dopamine transporter SPECT/CT and perfusion brain SPECT imaging in idiopathic basal ganglia calcinosis. AB - A case of idiopathic basal ganglia calcification in a 56-year-old woman with parkinsonism and cognitive impairment is described. The nigrostriatal dopaminergic pathway and regional cerebral blood flow were evaluated using dopamine transporter (DAT) brain single photon emission tomography combined with a low-dose x-ray computerized tomography transmission (hybrid SPECT/CT) and Tc 99m HMPAO brain perfusion SPECT study, respectively. DAT SPECT/CT imaging revealed a reduction in DAT binding in both striatum regions coinciding with bilateral calcifications in the basal ganglia. Brain perfusion scan showed hypoperfusion in basal ganglia regions, posterior parietal cortex bilaterally, left frontopolar and dorsolateral prefrontal cortex, and left temporal lobe. These findings correlated well with the clinical condition of the patient. Mineralization may play a critical role in the pathogenesis of neuronal degeneration. Cortical perfusion changes in patients may better explain the patient's altered cognitive and motor functions. PMID- 19542945 TI - Comparison of lung scintigraphy with multi-slice spiral computed tomography in the diagnosis of pulmonary embolism. AB - PURPOSE: To compare the diagnostic efficacy of lung perfusion scans combined with ventilation (V/Q) scans and/or chest radiography (CR) with contrast-enhanced multislice spiral CT pulmonary angiography (CTPA) in diagnosing pulmonary embolism (PE). MATERIALS AND METHODS: Eighty-two consecutive patients with suspected PE underwent CTPA, lung perfusion scan, and CR. Of them, 28 patients underwent V/Q scans. The final diagnosis was made using a composite reference test. RESULTS: The overall sensitivity and specificity were 89.2% and 92.1% for V/Q scan or perfusion scan combined with CR, and 97.3% and 97.4% for CTPA. For the 28 patients with V/Q scan, the sensitivity and specificity were 91.7% and 92.9% for V/Q scan, and 91.7% and 100.0% for CTPA. The segmental agreement rate between perfusion scan and CTPA was 69.5% (kappa = 0.30, P < 0.05). The perfusion scan revealed significantly more subsegmental abnormalities than CTPA (59 vs. 10, chi2 test, P < 0.05). CONCLUSIONS: V/Q scan, perfusion scan combined with CR and CTPA all show high efficacy in diagnosing PE. V/Q scan or perfusion scan combined with CR is as accurate as CTPA. PMID- 19542946 TI - Extensive FDG uptake in accessory muscles of respiration in a patient with shortness of breath. PMID- 19542947 TI - Tuberculous synovitis of the hip joint diagnosed by FDG PET-CT. PMID- 19542948 TI - Widespread tophaceous gout demonstrating avid F-18 fluorodeoxyglucose uptake. PMID- 19542949 TI - Mandibular lesion differential diagnoses in a patient with a previous history of locally advanced head and neck carcinoma. PMID- 19542950 TI - Hepatosplenic candidiasis imaged with F-18 FDG PET/CT. PMID- 19542951 TI - Simultaneously significant hepatic and mild splenic uptake of Tc-99m MDP resulting from Waldenstrom macroglobulinemia. PMID- 19542953 TI - Small bowel metastasis from lung cancer detected on FDG PET/CT. PMID- 19542952 TI - Breast angiosarcoma: FDG PET findings. PMID- 19542954 TI - F-18 FDG PET/CT imaging in a case of primary choriocarcinoma in the retroperitoneum. PMID- 19542955 TI - Suppression of F-18 fluorodeoxyglucose uptake in benign uterine leiomyomas with danazol. PMID- 19542956 TI - F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) and Tc-99m pertechnate scan findings of a patient with unilateral subacute thyroiditis. PMID- 19542958 TI - F-18 fluorodeoxyglucose positron emission tomography in a transitional meningioma. PMID- 19542957 TI - Ectopic ACTH syndrome due to occult bronchial carcinoid. PMID- 19542959 TI - Ventriculoperitoneal shunt catheter and cerebral spinal fluid infection initially detected by FDG PET/CT scan. PMID- 19542960 TI - Scintigraphic detection of I-125 seeds migration after permanent brachytherapy for prostate cancer: how far do seeds travel? PMID- 19542961 TI - F-18 FDG PET/CT scan in biliary cystadenocarcinoma. PMID- 19542962 TI - Tc-99m tetrofosmin SPECT in coronary cameral fistula. PMID- 19542963 TI - "Tight-fitting cloth"-like uptake of Ga-67 in a patient with psoriasis. PMID- 19542964 TI - Tc-99m labeled RBC scintigraphy of an inflammatory pseudotumor of the spleen. PMID- 19542965 TI - Tc-99m pertechnetate/sestamibi imaging in a case of recurrent parathyroid carcinoma with metabolic bone disorder. PMID- 19542967 TI - Current readings in nuclear medicine. PMID- 19542966 TI - Metastatic melanoma with multiple small bowel intussusceptions. PMID- 19542968 TI - Foreword. Healthy workplaces in acute care hospitals. PMID- 19542969 TI - Healthy, healthful, and healing environments: a nursing imperative. AB - The literature is replete with evidence about the effects of the work environment on nurses' stress levels, interdisciplinary collaboration, workload, job conflict, job satisfaction, and anticipated turnover. Healthcare leaders have been challenged by the American Association of Critical-Care Nurses (AACN), other professional organizations, and regulatory agencies to develop and sustain healthy work environments that support the professional practice of nursing. Magnet designation, the Beacon award, and other organizational structures and cultures led by authentic and transformational leaders have been the stimulus to ensure that workplaces are both healthy and healthful. The positive effect of healing environments on patient and provider outcomes has caused many healthcare leaders to strive to develop healing attributes within their philosophies of care and organizational cultural initiatives. PMID- 19542970 TI - Authentic leadership, organizational culture, and healthy work environments. AB - The purpose of this article is to showcase the relationship among authentic leadership, organizational culture, and healthy work environments using a stress and coping lens. A qualitative descriptive study was conducted to determine what situations contribute to nurse manager stress, what coping strategies they utilize, what health outcomes they report, and what decision-making processes they follow to address stressful situations in their roles. A purposive sample of 21 nurse managers employed at 3 US acute care hospitals completed a demographic questionnaire and 14-question interview incorporating components of the Critical Decision Method. A secondary analysis of the data was conducted to identify differences in nurse manager narratives based upon differences in the organizational cultures where the managers worked. Of the 21 nurse managers studied, differences were evident in the organizational cultures reported. Nurse managers working in the positive organizational cultures (n = 12) generally worked in healthy work environments and engaged in more authentic leadership behaviors. Conversely, nurse managers working in the negative organizational cultures (n = 9) worked in unhealthy work environments and reported less optimism and more challenges engaging in authentic leadership practices. Organizational culture and leadership matter in creating and sustaining healthy work environments. Nurse managers play a pivotal role in creating these environments, yet they need supportive structures and resources to more effectively execute their roles. PMID- 19542971 TI - An ethical framework for developing and sustaining a healthy workplace. AB - Acute and critical care nurses provide care that is intellectually challenging and physically demanding in environments that are often characterized by ethical concerns and dilemmas. As complex adaptive systems, these environments include nurses, physicians, social workers, chaplains, case managers, and administrators who interact together in situations of tremendous uncertainty and frequently in the face of professional or social disagreement about patient/family care, processes, and outcomes. Nurses' ability to identify and resolve concerns in such situations as part of a collaborative ethical climate is a significant retention factor and a hallmark of a healthy workplace. The purpose of this article is to describe a pluralist ethical framework that evolved from the formation of a healthy workplace and that may be integrated to sustain an ethical climate. Discussion of development of a healthy workplace employing participatory action research will be followed by a description of ethical theories supporting the framework and their relevance to creating and sustaining a healthy workplace and ethical practice environment. Finally, specific application examples of the ethical framework from nursing units' staff participatory action research studies will be presented. PMID- 19542973 TI - Teamwork: building healthier workplaces and providing safer patient care. AB - A changing healthcare landscape requires nurses to care for more patients with higher acuity during their shift than ever before. These more austere working conditions are leading to increased burnout. In addition, patient safety is not of the quality or level that is required. To build healthier workplaces where safe care is provided, formal teamwork training is recommended. Formal teamwork training programs, such as that provided by the MedTeams group, TeamSTEPPS (Team Strategies and Tools to Enhance Performance and Patient Safety), or participatory action research programs such as the Healthy Workplace Intervention, have decreased errors in the workplace, increased nurse satisfaction and retention rates, and decreased staff turnover. This article includes necessary determinants of teamwork, brief overviews of team-building programs, and examples of research programs that demonstrate how teamwork brings about healthier workplaces that are safer for patients. Teamwork programs can bring about these positive results when implemented and supported by the hospital system. PMID- 19542972 TI - Building organizational capacity for a healthy work environment through role based professional practice. AB - The professional practice of registered nurses (RNs) and their professional role competence are key variables that have an impact on quality and patient safety. Organizations in which RNs practice must have the capacity to fully support the professional role of those RNs in exercising their legitimate power derived through nurse licensing laws and professional standards and ethics. The interplay of individual RN practice and organizational practice, and measurement thereof, are the essence of organizational capacity. Two models are discussed that tie together the attributes of healthy workplace environments and provide the structure to guide and sustain organizational capacity. PMID- 19542974 TI - Case study: creating a healthy workplace in a surgical trauma intensive care unit. AB - This case study describes the implementation of the Parsons' Healthy Workplace Theory and Intervention in a surgical intensive care unit in a level 1 trauma facility. This intervention and change strategy was the impetus for the creation of a more positive work environment by developing and empowering staff. The process led to shared decision making and development of action planning teams that subsequently became unit-based committees. The committees have focused goals, action plans, and timelines for achieving those goals. Transforming care and the enculturation of shared decision making at the bedside is essential for improving quality of patient care and for recruitment and retention of nurses. PMID- 19542975 TI - HIPP lead to self-health: healthy infection prevention practices in intensive care units. AB - Healthy infection prevention practices (HIPP) include hand hygiene, respiratory etiquette, environmental cleanliness, and use of personal protective equipment. These healthy practices are most used to protect individuals against exposure to bacterial and viral infections in the workplace, as well as in the home. Most often these infection prevention behaviors in critical care units are promoted to protect the patient from healthcare-associated infections. Yet, these practices are just as important to the health of the critical care nurse. Self-health in the workplace is essential to creating a healthy workplace environment, which is the goal of many intensive care units today. The benefits of creating a healthy work environment are improvement of patient/nurse satisfaction and nurse retention. HIPP reduce the risk of the critical care nurse's exposure to microbial pathogens such as methicillin-resistant Staphylococcus aureus and influenza. Pathogens that cause infections may contaminate the hands, the clothing, equipment, and blood, putting the nurse at risk for unhealthy hands, unhealthy flora, and unhealthy blood. The intensive care nurse is encouraged to embrace HIPP to nurture self, as well as protect the patient. PMID- 19542979 TI - Bone disease in chronic kidney disease: unraveling the details. PMID- 19542977 TI - Healthy workplaces and ethical environments: a staff nurse's perspective. AB - Healthy workplaces contribute to improved patient safety and job satisfaction. In the development of healthy workplace cultures, various frameworks have been offered as strategy guides. There has also been a growing body of evidence that ethical climates and education can positively influence patient safety and job satisfaction. Within an ethical context, the American Association of Critical Care Nurses's Standards for Establishing and Sustaining Healthy Work Environments have been examined. It is proposed that the underpinnings of healthy workplace initiatives be built upon an ethical foundation that is apparent to staff nurses. The benefit of incorporation of an ethical context into workplace change is discussed. Evidence to support the proposal is cited and examples are given. PMID- 19542980 TI - Podocyte polarity signalling. AB - PURPOSE OF REVIEW: The glomerular filtration barrier is a unique structure characterized by a specialized three-dimensional framework of podocytes. This review is aimed at describing the latest advances made in the understanding of polarity signalling pathways regulating the formation and the maintenance of the complex podocyte architecture. RECENT FINDINGS: Podocytes are composed of a large cell body that extends primary and secondary processes. An apicobasal polarity axis allows for podocyte orientation between the urinary space and the glomerular basement membrane. Recent studies document that conserved polarity protein complexes such as the partitioning defective 3 (Par3), partitioning defective 6 (Par6) and atypical protein kinase C (aPKC) complex are essential regulators of podocyte morphology. Glomerular development, slit diaphragm targeting and apicobasolateral distribution of molecules seem to be tightly regulated by these polarity signalling pathways. SUMMARY: Accumulating evidence indicates that conserved polarity protein complexes are essential for normal podocyte morphology and differentiation. The diseased podocyte, which typically presents with foot process effacement, might require these molecular guideposts when recovering from stress and when restoring normal podocyte morphology. PMID- 19542981 TI - Current world literature. PMID- 19542982 TI - Physical activity and risk of endometrial cancer: an Italian case-control study. AB - Most epidemiological studies on the relationship between physical activity and endometrial cancer found risk reductions of about 25-30% among most active women, but results were not consistent among studies. A multicentric case-control study was conducted in Italy between 1992 and 2006. Cases were 454 women with incident, histologically confirmed endometrial cancer and controls were 908 women admitted to hospital for acute non-neoplastic, nonhormonal conditions. Multivariate odds ratios (ORs) and the corresponding 95% confidence intervals were obtained after allowance for major potential confounding factors. The ORs of endometrial cancer for women in the highest level of occupational physical activity (compared with the lowest) were 1.69, 1.33, 1.17 and 0.82, respectively, at ages 12, 15-19, 30 39 and 50-59 years, with no trend in risk at any age. The corresponding ORs for leisure-time physical activity were 0.82, 0.78, 1.12 and 0.97. The risk of endometrial cancer for each level of occupational physical activity at age 30-39 years was not significantly heterogeneous across strata of age at diagnosis, body mass index, menopausal status and education. These findings do not support a strong relationship between physical activity and endometrial cancer risk. PMID- 19542983 TI - The efficacy of pregabalin and benzodiazepines in generalized anxiety disorder presenting with high levels of insomnia. AB - The objective of this study was to assess the impact of high levels of insomnia on response to pregabalin (PGB) in patients with generalized anxiety disorder (GAD). Pooled data were analyzed from six double-blind, placebo-controlled, 4- to 6-week trials of outpatients who met the Diagnostic and Statistical Manual of Mental Disorders (fourth edition) criteria for GAD with a minimum Hamilton Rating Scale for Anxiety (HAM-A) score = 18. Response was evaluated for three fixed-dose PGB groups (150, 300-450, 600 mg/day), and for a benzodiazepine group (alprazolam or lorazepam). A 'high-insomnia' subgroup was defined by a baseline HAM for Depression (HAM-D) insomnia factor score greater than 3 (maximum = 6). At baseline, 1002 (54%) patients met the criteria for the high-insomnia subgroup, and 852 (46%) for the low-insomnia subgroup. Mean baseline HAM-A scores were 1-2 points higher in high-insomnia versus low-insomnia patients. In high-insomnia patients, PGB produced significantly greater improvement in HAM-A total scores at last observation carried forward endpoint on 300-450 mg (-13.1+/-0.6) and 600 mg (-11.2+/-0.5) dose groups compared with placebo (-8.3+/-0.5; P<0.0001 for both comparisons); the improvement on PGB 150 mg was not significant (-9.9+/-0.7; P = 0.051). Improvement was significant in the benzodiazepine group (-11.0+/-0.6; P<0.0001). In the high-insomnia subgroup, treatment with PGB significantly (P<0.001) improved the HAM-D insomnia factor scores on both the 300-450 mg ( 2.73) and 600 mg (-2.35) doses, and on benzodiazepines (-2.52) compared with placebo (-1.51); improvement on PGB 150 mg (-1.69) was not significant. Rates of treatment-emergent insomnia were lower on PGB compared with placebo in both the high- and low-insomnia subgroups. In conclusion, PGB was well tolerated, and improved overall anxiety symptoms, while specifically improving insomnia in patients with GAD presenting with high levels of concurrent insomnia. PMID- 19542984 TI - Fibrinogen, genes, and arterial stiffness. PMID- 19542985 TI - Metabolic syndrome, insulin resistance and oxidative stress: adding insights to improve cardiovascular prevention. PMID- 19542986 TI - Large artery stiffness and microalbuminuria: a causal relationship? PMID- 19542987 TI - Shortness of breath, prescription of bronchodilators and the risk of myocardial infarction. PMID- 19542988 TI - Sunitinib-aggravated hypertension. PMID- 19542989 TI - Increased burden of high blood pressure and related risk factors among participants of World Hypertension Day 2008 in Cameroon. PMID- 19542990 TI - Cardiovascular risk in young adults. PMID- 19542991 TI - Stroke risk and antihypertensive drug therapy. PMID- 19542994 TI - The trigemino-cardiac reflex: an update of the current knowledge. AB - The trigemino-cardiac reflex (TCR) is clinically defined as the sudden onset of parasympathetic activity, sympathetic hypotension, apnea, or gastric hypermotility during central or peripheral stimulation of any of the sensory branches of the trigeminal nerve. Clinically, the TCR has been reported to occur during craniofacial surgery, manipulation of the trigeminal nerve/ganglion and during surgery for lesion in the cerebellopontine angle, cavernous sinus, and the pituitary fossa. Apart from the few clinical reports, the physiologic function of this brainstem reflex has not yet been fully explored. The manifestation of the TCR can vary from bradycardia and hypotension to asystole. From the experimental findings, the TCR represents an expression of a central reflex leading to rapid cerebrovascular vasodilatation generated from excitation of oxygen-sensitive neurons in the rostral ventro-lateral medulla oblongata. By this physiologic response, the systemic and cerebral circulations may be adjusted in a way that augments cerebral perfusion. This review summarizes the current state of knowledge about TCR. PMID- 19542995 TI - Pulmonary function after surgery for congenital atlantoaxial dislocation: a comparison with surgery for compressive cervical myelopathy and craniotomy. AB - Deterioration of pulmonary function after surgery for congenital atlantoaxial dislocation (AAD) has been documented in a few studies. We proposed that this deterioration in AAD is much higher than what can be expected after a surgical procedure under general anesthesia or what occurs after any surgery on the cervical spine. To test this hypothesis, we recorded forced vital capacity (FVC), forced expiratory ratio (FEV 1.0), forced expiratory flow (FEF 25%-75%) and muscle power in the extremities in 25 patients undergoing surgical correction of AAD (AAD group), 29 patients undergoing surgery for compressive cervical spine lesions (cervical spine group) and 20 patients undergoing craniotomy for an intracranial lesion (craniotomy group). The observations were made before surgery and on postoperative days 1 and 7. The demographic characters were comparable among the 3 groups. All patients underwent an uneventful surgery and their trachea was extubated in the operating room. There was no decrease in the muscle power in the postoperative period in any of the groups. A significant decrease in FVC (expressed as percentage of the predicted value) was seen postoperatively in all the 3 groups. The reduction of FVC was significantly different among the groups, with the AAD group having the lowest values (P<0.001). The FVC values in the AAD group were 74.6+/-19.6%, 49.6+/-17.7%, 64.0+/-20.8% at baseline, on postoperative days 1 and 7, respectively (P<0.001). Postoperative change in forced expiratory ratio was also significantly different among the groups (P=0.03). A significant difference was found between the AAD and cervical spine group (89.8+/-8.3%, 88.2+/-17.6%, 89.3+/-9.8% in the AAD group and 95.5+/-20.5%, 78.4+/-13.4%, 72.7+/-19.1% in the cervical spine group at baseline and on postoperative days 1 and 7, respectively, P<0.05). FEF 25%-75% changes were also significantly different among the groups (P<0.001). The decrease in the AAD and cervical spine groups was significantly higher than that in the craniotomy group (P<0.001). In conclusion, during the first week after surgery, deterioration of pulmonary function in the AAD group is significantly different from that seen in patients undergoing surgery for compressive cervical lesions or craniotomy for a cerebral lesion. The data imply the need for special attention to respiratory function in patients operated for AAD in the postoperative period. PMID- 19542996 TI - Sevoflurane provides better recovery as compared with isoflurane in children undergoing spinal surgery. AB - Rapid recovery is desirable in pediatric neurosurgical patients to obtain an early neurologic assessment. We compared the recovery characteristics of 2 commonly used anesthetic agents, sevoflurane and isoflurane, under bispectral index-guided anesthesia in children undergoing spinal surgery. Eighty children who underwent surgery for occult spinal dysraphism at the lumbar and sacral level were randomized to anesthesia with sevoflurane or isoflurane in oxygen and nitrous oxide. Anesthesia depth was guided by a bispectral index monitor kept between 40 and 60. In addition to time at emergence, extubation and discharge, recovery (modified Aldrete) score, and hemodynamics were compared. The 2 groups did not differ significantly with respect to demographics, duration of surgery and anesthesia, and intraoperative hemodynamic changes. Extubation (6.4+/-3.3 vs. 10.7+/-4.6) and emergence (7.8+/-3.4 vs. 12.8+/-5.6) times (minutes) were significantly shorter with sevoflurane (P<0.001). Time (minutes) to achieve full Aldrete (modified) scores was less with sevoflurane (13.9+/-5.3 vs. 20.3+/-6.5) than isoflurane (P<0.001). However, the time (minutes) to achieve discharge criteria from postanesthesia care unit (140.7+/-49.3 vs. 146+/-43.3) and first dose of postoperative analgesic (60+/-24.1 vs. 72+/-33.4) in addition to incidence of postoperative agitation were similar in both groups (P>0.05). Sevoflurane results in an earlier recovery and assessment of modified Aldrete score when compared with isoflurane. PMID- 19542998 TI - A single dose of esmolol blunts the increase in bispectral index to tracheal intubation during sevoflurane but not desflurane anesthesia. AB - Activation of the peripheral nerve system by endotracheal intubation is accompanied by an increase in bispectral index (BIS). Esmolol produces a dose dependent attenuation of the adrenergic response to endotracheal intubation. Desflurane increases sympathetic nerve activity and plasma norepinephrine relative to sevoflurane. The authors hypothesized that esmolol might blunt the BIS response to endotracheal intubation more during sevoflurane anesthesia than desflurane anesthesia. In this double blind, randomized study, after the induction of anesthesia, patients were mask-ventilated with either sevoflurane or desflurane (end-tidal 1 minimum alveolar concentration) and received normal saline or esmolol (0.5 mg/kg) 1 minute before intubation (sevoflurane-control, sevoflurane-esmolol, desflurane-control, and desflurane-esmolol groups, n=20/group). BIS, mean arterial pressure, and heart rate were measured before the induction of anesthesia (awake), before esmolol injection (time point -1), immediately before intubation (time point 0), and every minute for 5 minutes after tracheal intubation (time point 1 to 5). Compared with preintubation, esmolol attenuated the increase in BIS at 1 minute after intubation during sevoflurane anesthesia (5.1% for esmolol and 31.7% for control) but not during desflurane anesthesia (28.6% for esmolol and 30.8% for control). Mean arterial pressure and heart rate increased after intubation in all groups but the changes were greater in the control groups than the esmolol groups. In conclusion, a single dose of esmolol blunted the increase in BIS to tracheal intubation during sevoflurane but not desflurane anesthesia. PMID- 19542997 TI - Early postoperative cognitive recovery and gas exchange patterns after balanced anesthesia with sevoflurane or desflurane in overweight and obese patients undergoing craniotomy: a prospective randomized trial. AB - Overweight and obese patients are at especially high risk for delayed awakening after general surgery. Whether this risk also applies to cerebral neurosurgical procedures remains unclear. This study evaluated early postoperative cognitive recovery and gas exchange patterns, after balanced anesthesia with sevoflurane or desflurane, in overweight and obese patients undergoing craniotomy for supratentorial expanding lesions. Fifty-six patients were consecutively enrolled, and randomly assigned to 1 of 2 study groups to receive balanced anesthesia with sevoflurane or desflurane. Cognitive function was evaluated with the Short Orientation Memory Concentration Test and the Rancho Los Amigos Scale and gas exchange patterns (pH, PaO2, and PaCO2) were recorded in all patients at 5 time points: preoperatively and postoperatively, after patients reached an Aldrete score >or=9, at 15, 30, 45, and 60 minutes. Preoperative cognitive status was similar in the 2 treatment groups. Early postoperative cognitive recovery was more delayed and Short Orientation Memory Concentration Test scores at 15 and 30 minutes postanesthesia were lower in patients receiving sevoflurane-based anesthesia than in those receiving desflurane-based anesthesia (21.5+/-3.5 vs. 14.9+/-3.5) (P<0.005) and (26.9+/-0.7 vs. 21.5+/-1.4) (P<0.005), and the postoperative Rancho Los Amigos Scalegrade 8 showed a similar trend (25/28 patients 89% vs. 8/28 patients 28% (P<0.005) and 28/28 patients (100% vs. 13/28 patients 46%) (P<0.005). Similarly, gas-exchange analysis showed higher PaCO2 at 15 and 30 minutes and lower pH up to 45 minutes postextubation in patients receiving sevoflurane-based anesthesia. In overweight and obese patients undergoing craniotomy desflurane-based anesthesia allows earlier postoperative cognitive recovery and reversal to normocapnia and normal pH. PMID- 19542999 TI - Evaluation of the FloTrac uncalibrated continuous cardiac output system for perioperative hemodynamic monitoring after subarachnoid hemorrhage. AB - Early hemodynamic assessment is of particular importance for adequate cerebral circulation in patients with aneurysmal subarachnoid hemorrhage (SAH), but is often precluded by the invasiveness and complexity of the established cardiac output determination techniques. We examined the utility of an uncalibrated arterial pressure-based cardiac output monitor (FloTrac) for intraoperative and postoperative hemodynamic management after SAH. In 16 SAH patients undergoing surgical clipping, arterial pulse contour cardiac index, and stroke volume variation (SVV) were analyzed via the radial FloTrac system. The hemodynamic values after induction of anesthesia until 12 hours after surgery were compared with reference transpulmonary thermodilution cardiac index (TPCI), calibrated pulse contour CI, and global end-diastolic volume index determined by the PiCCO system and central venous pressure. Arterial pulse contour cardiac index underestimated CI as overall bias+/-SD of 0.57+/-0.44 L/min/m2 and 0.54+/-0.46 L/min/m2 compared with TPCI and calibrated pulse contour CI, resulting in a percentage error of 24.8% and 26.6%, respectively. Subgroup analysis revealed a percentage error of 29.3% for values obtained intraoperatively and 20.4% for values measured under spontaneously breathing after tracheal extubation. Better prediction of cardiac responsiveness to defined volume loading for increasing stroke volume index >10% was observed for SVV under mechanical ventilation with greater area under the receiver operating characteristics curve than that for global end-diastolic volume index or central venous pressure. These data suggest that the FloTrac underestimates the reference CI, and is not as reliable as transpulmonary thermodilution for perioperative hemodynamic monitoring after SAH. SVV is considered to be an acceptable preload indicator under mechanical ventilation. PMID- 19543000 TI - A randomized, double-blinded comparison of ondansetron, granisetron, and placebo for prevention of postoperative nausea and vomiting after supratentorial craniotomy. AB - Postoperative nausea and vomiting (PONV) are frequent and distressing complications after neurosurgical procedures. We evaluated the efficacy of ondansetron and granisetron to prevent PONV after supratentorial craniotomy. In a randomized double-blind, placebo controlled trial, 90 adult American Society of Anesthesiologists I, II patients were included in the study. A standard anesthesia technique was followed. Patients were divided into 3 groups to receive either placebo (saline), ondansetron 4 mg, or granisetron 1 mg intravenously at the time of dural closure. After extubation, episodes of nausea and vomiting were noted for 24 hours postoperatively. Statistical analysis was performed using chi2 test and 1-way analysis of variance. Demographic data, duration of surgery, intraoperative fluids and analgesic requirement, and postoperative pain (visual analog scale) scores were comparable in all 3 groups. It was observed that the incidence of vomiting in 24 hours, severe emetic episodes, and requirement of rescue antiemetics were less in ondansetron and granisetron groups as compared with placebo (P<0.001). Both the study drugs had comparable effect on vomiting. However, the incidence of nausea was comparable in all 3 groups (P=0.46). A favorable influence on the patient satisfaction scores, and number needed to prevent emesis was seen in the 2 drug groups. No significant correlation was found between neurosurgical factors (presence of midline shift, mass effect, pathologic diagnosis of tumor, site of tumor) and the occurrence of PONV. We conclude that ondansetron 4 mg and granisetron 1 mg are comparably effective at preventing emesis after supratentorial craniotomy. However, neither drugs prevented nausea effectively. PMID- 19543001 TI - Effect of tetracaine-induced spinal anesthesia on pial microcirculation in pentobarbital anesthetized rats. AB - Local anesthetic is administrated intrathecally to produce spinal anesthesia. This study examines the effect of spinal anesthesia induced by intrathecal tetracaine on cerebral pial microcirculation in rats. We monitored changes in the mean arterial pressure, internal diameter (ID) of the pial arteriole, intracranial pressure, and red blood cell velocity. The regional cerebral blood flow was calculated from the product of cross sectional area and red blood cell velocity. To induce spinal anesthesia, tetracaine was administered via a polyethylene tube at the L4-5 intervertebral space after laminectomy. Arterial blood pressure was monitored via a catheter in the femoral artery. A left parietal craniotomy with an encapsulated cranial window was prepared for biomicroscopy. Change in the arteriolar ID was measured by image shearing. Red blood cell velocity was measured by the dual-slit photometric method and correlation technique. Three levels of spinal anesthesia were evaluated. Tetracaine at 0.1 and 0.2 mg/kg, which produced T10 and T6 sensory block, respectively, did not cause significant change of ID of the pial arteriole and red blood cell velocity. Tetracaine (0.3 mg/kg) produced total spinal block, resulting in a significant decrease of the calculated cerebral blood flow and caused brief pial arteriolar vasodilation followed by vasoconstriction. Our results show that total spinal anesthesia with tetracaine causes significant changes in rat cerebral microcirculation. PMID- 19543002 TI - The neuroprotective effects of oxaloacetate in closed head injury in rats is mediated by its blood glutamate scavenging activity: evidence from the use of maleate. AB - INTRODUCTION: Treatment with oxaloacetate after traumatic brain injury has been shown to decrease blood glutamate levels and protect against the neurotoxic effects of glutamate on the brain. A number of potential mechanisms have been suggested to explain oxaloacetate-induced neuroprotection. We hypothesize that the primary mechanism by which intravenous oxaloacetate provides neuroprotection is by activation of the blood glutamate-scavenging enzyme glutamate-oxaloacetate transaminase, increasing thereby the driving force for the efflux of excess glutamate from brain interstitial fluids into blood. If so, coadministration of maleate, a glutamate-oxaloacetate transaminase-blocker is expected to prevent the neuroprotective effects of oxaloacetate. MATERIALS AND METHODS: A neurological severity score (NSS) was measured 1 hour after closed head injury (CHI) in rats. Then, rats received 30 microL/min/100 g infusion of saline, or 1 mmol/100 g solution of oxaloacetate, maleate, or a mixture of oxaloacetate and maleate. NSS was reassessed at 24 and 48 hour after CHI. Blood glutamate and glucose levels were measured at 0, 60, 90, and 120 minutes. RESULTS: NSS improved significantly at 24 hour (P<0.001) and 48 hour (P<0.001) only in the rats treated with oxaloacetate. Blood glutamate decreased significantly in the oxaloacetate-treated group at 90 minute (at the conclusion of oxaloacetate administration) (P<0.00001), but not in the control, maleate or oxaloacetate+maleate groups. A strong correlation r2=0.86 was found to exist between the percent decrease in blood glutamate levels and percent improvement in NSS. DISCUSSION: The results of this study demonstrate that the primary mechanism by which oxaloacetate provides neuroprotective activity after CHI is related to its blood glutamate scavenging activity. Management of blood glutamate concentration may have important implications in the treatment of acute brain conditions, including CHI and stroke. PMID- 19543003 TI - Spectral analysis of heart rate variability during asleep-awake craniotomy for tumor resection. AB - Anesthesia during asleep-awake craniotomy should provide adequate analgesia and sedation whereas permitting language testing. In this work, we used the analysis of heart rate variability (HRV) to quantify the sympatho-vagal balance and better evaluate patient's stress response during asleep-awake craniotomy. Patients admitted to our hospital for tumor resection with language testing were studied (n=21, age range: 22 to 53 y ). Heart rate and systolic arterial blood pressure were collected at 5 time points: T1: preanesthesia; T2: dura mater opening; T3: cortical mapping; T4: subcortical mapping; T5: dura mater suturing. Patients were anesthetized with propofol/remifentanil infusion and ventilated via laryngeal mask during T2, but were awakened for language testing at T3 and T4, and resedated with remifentanil during T5. At each time point, HRV was analyzed by power spectrum analysis: overall variance, very low frequency (VLF), low frequency (LF) and high frequency (HF) powers, and LF/HF ratio (an index of prevalence of sympathetic over parasympathetic tone) were derived. A significant increase in both heart rate and systolic arterial blood pressure was observed from time point T3 through T5 (P<0.05, compared with T1). HRV analysis revealed that the LF/HF ratio progressively increased to reach values during T4 that were significantly higher than preanesthesia values (P<0.05). During T5, LF/HF ratio returned to preanesthesia level. HRV analysis confirmed the presence of moderate intraoperative stress response, indicating a significant increase in the LF/HF ratio during the awake phases. This information might help in tailoring the protocol and the duration of awake phase according to the individual autonomic response. PMID- 19543004 TI - Hemodynamic changes after administration of mannitol measured by a noninvasive cardiac output monitor. AB - Mannitol is the most commonly used hyperosmotic agent in neurosurgery. Being an agent that increases intravascular volume by withdrawing water from the brain, it may cause significant changes in stroke volume (SV), cardiac output (CO), systemic vascular resistance and blood pressure. In this study, we monitored the hemodynamic changes in response to a single dose of mannitol by using a noninvasive CO monitor based on the thoracic electrical bioimpedance technique, in patients undergoing craniotomy. Eleven adult patients undergoing elective craniotomy received mannitol 1.0 g/kg 15 minutes before dural opening. The following hemodynamic variables were recorded: heart rate, systolic blood pressure, diastolic blood pressure, SV, CO, and cardiac index. The measurements were made before the administration of mannitol, at 1, 2, 5, 10, 15, 20, 25, 30, 35, 40, and 45 minutes after the termination of the mannitol infusion. Urine output was measured at 10, 20, 30, 40, 50, 60, 90, and 120 minutes after termination of the mannitol infusion. Heart rate values from 25 to 45 minutes were significantly lower compared with the premannitol values (P<0.05). All the postmannitol systolic blood pressure values were significantly lower than the premannitol value (P<0.05). SV increased significantly for 15 minutes after administration of mannitol (P<0.05). SV at 45 minutes was significantly lower than that from 1 to 30 minutes (P<0.05). Cardiac index also showed a similar change with a significant increase at 1 to 10 minutes and a decrease at 40 to 45 minutes compared with 1 to 15 minutes.The rate of urine secretion was higher during the first 10 minutes (40+/-15 mL/kg/ h) than during the rest of the study period. The overall fluid balance at the end of 120 minutes was -370+/-987 mL. In this study using noninvasive measurement of CO by thoracic bioimpedance plethysmography during craniotomy, a single bolus dose of mannitol 1.0 g/kg caused a significant but short duration changes in the hemodynamic variables. The changes in SV, and CO, lasted for only 15 minutes after the infusion. PMID- 19543008 TI - Electrocardiographic changes due to vagosympathetic coactivation during the trigeminocardiac reflex. PMID- 19543005 TI - Functional magnetic resonance imaging (fMRI) in children sedated with propofol or midazolam. AB - Magnetic resonance imaging (MRI) requires patient immobility and children generally need to be sedated. The ideal sedative agent for functional MRI (fMRI) should only minimally hamper the neurophysiologic effect of the administered sensorial stimulation. This study compares the effect of propofol and midazolam on the fMRI auditory activation pattern in children. Fourteen children in the 3 to 7 year age group without neurologic or auditory deficits were randomly assigned to receive propofol or midazolam for sedation during auditory fMRI. Two patients in the midazolam group were excluded due to positive baseline MRIs. The children were stimulated using a passive listening task. The fMRI signal was modeled using various functions (hemodynamic response function, temporal derivative, and dispersion derivative) to check for the differing temporal characteristics of the signal between the groups. Patients in the propofol group showed activation only in the primary auditory cortex and exhibited a pattern more similar to that of nonsedated adults. Patients in the midazolam group exhibited a more complex pattern, presenting activation areas other than the primary auditory cortex; a delay in the functional response and higher duration variability were also observed. Our sample sizes are too small to derive a conclusive inference. Our preliminary study encourages the hypothesis that propofol is preferable to midazolam to maintain sedation in 3 to 7-year-old children during auditory fMRI because it facilitates the elicitation of a more focused auditory cortical activation pattern with less temporal and spatial dispersion. PMID- 19543010 TI - Trauma is a team sport. PMID- 19543014 TI - Utilizing trauma nurse practitioners to decrease length of stay. AB - This article describes a nurse practitioner model utilized to decrease the length of stay and improve the quality of discharge planning for hospitalized trauma patients between 1999 and 2006. An observational method employing nurse practitioners to decrease length of stay for the trauma population during these years is described. Adding nurse practitioners to the trauma team has resulted in decreasing the length of stay in all 4 of the injury severity score groups. Adding nurse practitioners to the trauma team provides a core member in a revolving trauma service. Consequently, length of stay and discharge planning have been positively impacted. PMID- 19543015 TI - "Helping them understand": nurses' caring behaviors as perceived by family members of trauma patients. AB - This qualitative study explored family members' perceptions of nurses' caring behaviors. Semistructured interviews were conducted with 10 family members of moderately to severely injured trauma patients cared for at a level II trauma center. The dominant behavior identified as being caring was explaining what was going on and interpreting medical jargon. Appearing hurried and abrupt was noted as behavior that was not caring. In a highly stressful situation such as trauma, the interpersonal relationship the nurse develops with the family influences the perception of the care received. PMID- 19543016 TI - Management of embedded foreign body: penetrating stab wound to the head. AB - Penetrating craniocerebral trauma is an injury in which a projectile violates the skull but does not exit. The significance of penetrating injuries to the head depends largely on the circumstances of the injury, the velocity of impact, and attributes of the projectile. While most penetrating head injuries are caused by firearms, lower-velocity mechanisms of penetrating brain injury present unique challenges for the multidisciplinary team involved with the delivery of care. Appropriate management can lead to optimal outcomes and limit secondary brain injury. PMID- 19543017 TI - Senior lifestyles and injury prevention: evaluating the effectiveness of an injury prevention program for older adults. AB - The purpose of this multicenter, before-and-after observational study was to determine whether a short educational intervention was associated with improvement in self-reported safety behavior in older adults. We developed 4 original injury prevention presentations with companion testing materials: Motor Vehicle Safety, Fall Prevention, Pedestrian Safety, and Home Safety. Participants also completed pre-post Short Form Health Survey Instrument (SF-12) quality-of life surveys. Of 414 participants, 226 completed follow-up testing and SF-12 surveys, for a 54.6% response rate. Those who completed either Pedestrian or Home Safety program showed no significant changes (P > .05) in either test scores or SF-12, and they comprised 61.9% of the final sample. Participants in the Motor Vehicle Safety and Fall Prevention programs accounted for 38.1% of the final sample and did show significant improvements between pre-post test scores. Only Fall Prevention participants showed significant differences in pre-post SF-12 scores. In the Fall Prevention group, numerous SF-12 subscores from the initial survey were significantly inversely correlated with pretest scores, and improvements in some SF-12 subscores correlated with improvements in test scores. Findings from the Fall Prevention group suggest that seniors with quality-of-life limitations may be aware of their increased risk and more willing to make changes to enhance safety. Further study is needed because many questions regarding optimal approaches to injury prevention in the aging demographic remain unanswered. PMID- 19543018 TI - Older adults with traumatic rib fractures: an evidence-based approach to their care. AB - It is expected that over the next decade the population of older adults in the United States will increase dramatically. As the older adult population increases, the number of older adults involved in traumatic accidents is also expected to climb. The older population is at an increased risk for complications and poor outcomes following trauma. Practitioners caring for these older adults will need to use evidence-based practice guidelines in an attempt to improve outcomes. This article provides a clinical guideline for the assessment and management of pain in older adults with traumatic rib fractures, and an approach for pain assessment, which includes the use of the numeric rating scale as well as incentive spirometry. The modalities used for pain management include epidural analgesia, paravertebral analgesia, patient controlled analgesia, and the use of oral opioids. PMID- 19543019 TI - Wheelchair falls: 5 years of data from a level I trauma center. AB - Falling from a wheelchair can result in a serious injury. Our report details injuries sustained by 30 individuals who fell from a wheelchair and presented to our level I trauma center over 5 years. Most fall victims (60%) were older than 65 years. The most common injuries were traumatic brain injury, femur fractures, and concussion. The most serious injuries were traumatic brain injuries and a vertebral fracture with resultant spinal cord injury. In the trauma setting, practitioners discharging patients using wheelchairs should be aware of this mechanism of injury and should provide education to ensure proper fit and use of the device. PMID- 19543020 TI - Suicidal behavior after a traumatic event. AB - This article reviews the literature on the psychosocial and psychiatric consequences (such as a diagnosis of posttraumatic stress disorder and elevated suicide risk) for a range of traumatic life experiences, including military combat, criminal victimization, interpersonal and domestic violence, child abuse and neglect, sexual retraumatization, natural disasters, torture, and the Holocaust. Possible mechanisms mediating the relationship between trauma and suicide are presented along with suicide risk assessment implications for trauma nurses. PMID- 19543021 TI - Danger, Will Robinson, danger!! PMID- 19543022 TI - Promoting research in clinical practice: strategies for implementing research initiatives. AB - Promoting the use and conduct of research in clinical practice is an essential component in ensuring best practices and promoting positive outcomes for patients. Yet, it is often difficult for nurses in clinical practice to devote time to research activities because the daily demands of practice often take precedence. This article reviews key strategies for implementing research in clinical practice. Examples of ongoing initiatives being conducted at a variety of clinical settings are highlighted to showcase strategies for implementing clinically focused research projects. PMID- 19543024 TI - Tough love for nursing. PMID- 19543031 TI - Reducing the cost of catheter-related bloodstream infections. PMID- 19543033 TI - . . . About organ donation. PMID- 19543034 TI - HIV treatment update released. PMID- 19543035 TI - Unlocking the secrets of locked-in syndrome. PMID- 19543037 TI - Severe sunburn. PMID- 19543038 TI - How to respond when lightning strikes. PMID- 19543039 TI - Getting to the root of your patient's back pain. PMID- 19543040 TI - Legally speaking...when can you say NO? PMID- 19543042 TI - A dose of compassion: it would take all of my skills and creativity to calm Walter. PMID- 19543044 TI - Looking into problems with transvaginal surgical mesh. PMID- 19543045 TI - Understanding headache pain in children. PMID- 19543046 TI - On the lookout for listeriosis. PMID- 19543047 TI - Combination drugs... PMID- 19543048 TI - Documenting gastric lavage. PMID- 19543049 TI - Hypokalemia. PMID- 19543050 TI - Psychiatric care of patients undergoing organ transplantation. AB - Transplantation provides life-saving therapy to critically ill patients with end stage organ failure. We found that the specialized knowledge of mental health clinicians and physicians, committed to the evaluation and management of transplant recipients, is essential for the optimal care of these patients who require a life-long regimen of immunosuppressive medication and dependency on the transplant team. Critical issues requiring psychiatric input are frequently encountered in the following areas (1) patient selection and informed consent; (2) recognition and pretransplant treatment of psychiatric disorders; (3) substance or alcohol abuse; (4) compliance with medical regimens; (5) posttransplant psychosis; and (6) immunosuppressive/psychiatric drug-drug interactions. This review focuses on the priorities in psychological and psychiatric care of these unique patients. PMID- 19543051 TI - Suppression of the obliteration process by ventilation in a mouse orthotopic tracheal transplantation model. AB - BACKGROUND: Obliterative airway disease (OAD) has been a major obstacle to long term survival after lung or tracheal transplantations, but the role of airflow has not been examined in the orthotopic or the heterotopic tracheal transplantation models. METHODS: Sixty mice were assigned to two experimental groups. Two C57BL/6 tracheal segments were surgically prepared and then orthotopically transplanted into allogeneic BALB/c recipients. In group A mice, both segments were left patent, whereas in group B mice, one of the donor tracheas was occluded with a silk knot to obstruct airflow. Histology, quantitative OAD measurements, electron microscopy, immunohistochemical staining, and apoptosis measurement of the epithelium were performed. RESULTS: Gross examination at harvest showed patent lumens of all tracheal segments. Group A allografts (ventilating tracheas) showed a markedly higher proportion of ciliated epitheliums and less lymphocyte infiltration in the lamina propria, whereas the epithelium appeared metaplastic in group B, with a higher proportion of flattened attenuated epithelium and loss of the normal ciliate architecture. Quantitative morphometric measurements suggested more prominent OAD manifestations in the nonventilating allografts of group B than were present in group A, although recipient-derived epithelium was observed in all allografts under immunohistochemical staining. The apoptotic indexes of the epithelium were 12.1% in allografts with adequate ventilation (group A) and 66.2% in ventilation occluded allotracheas (group B). CONCLUSIONS: OAD severity and the epithelial repopulation process are closely related to the physiologic environment of airflow. Further research is warranted to explore the underlying mechanisms of this phenomenon. PMID- 19543052 TI - Mesenchymal stem cells prolong composite tissue allotransplant survival in a swine model. AB - BACKGROUND: This study investigated whether mesenchymal stem cells (MSCs) combined with bone marrow transplantation (BMT), irradiation, or short-term immunosuppressant therapy could prolong composite tissue allotransplant survival in a swine hind-limb model. METHODS: Heterotopic hind-limb transplantation was performed in outbred miniature swine. Group I (n=5) was the untreated control. Group II (n=3) received MSCs alone (given on days -1, +3, +7, +14, +21). Group III (n=6) received cyclosporine A (CsA days 0 to +28). Group IV (n=4) received preconditioning irradiation (day -1), BMT (day +1), and CsA (days 0 to +28). Group V (n=5) received irradiation (day -1), BMT (day +1), CsA (days 0 to +28), and MSCs (days +1, +7,+14). The expression and localization of CD4/CD25 T cells and MSCs were assessed using flow cytometry and immunohistochemistry. RESULTS: The allografts survival with MSCs alone revealed a significant prolongation, when compared with the controls (P=0.02). Allografts with CsA treatment exhibited delayed rejection. Irradiation and BMT-CsA treatment revealed no significant allograft survival benefit when compared with the CsA treatment group, but graft versus-host disease (GVHD) was evident. However, combination of MSCs-BMT-CsA treatment demonstrated significant prolongation of allograft survival (>200 days, P<0.001) and no signs of GVHD with the lowest degree of rejection in the allo skin and interstitial muscle layers. The CD4/CD25 regulatory-like T-cell expression in the circulating blood and allo-skin significantly increased in the MSC-BMT-CsA group. Examination of bromodeoxyuridine-labeled MSCs revealed donor MSC engraftment into the recipient and donor skin and the recipient liver parenchymal tissue. CONCLUSION: These results suggested that the regulatory activity of MSCs on T cells and GVHD might contribute to significant prolongation of composite tissue allotransplant survival in the MSC-BMT-CsA treatment. PMID- 19543053 TI - Interaction of programmed death-1 and programmed death-1 ligand-1 contributes to testicular immune privilege. AB - BACKGROUND: Immune responses are tempered in immunologically privileged sites including the testis. Previous studies have shown that islet transplantation in the testis significantly prolongs islet allograft survival. However, mechanisms underlying testicular immune privilege and intratesticular allograft survival remain unclear. METHODS: Allogeneic murine islets were transplanted in the testis. Programmed death-1 ligand-1 (PD-L1) expression was detected by immunohistochemstry and real-time polymerase chain reaction. Infiltrating T-cell proliferation was measured by bromodeoxyuridine uptakes, whereas their apoptosis was quantified by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling methods. Transgenic T cells were used to track allospecific memory T cell generation. RESULTS: We found that programmed death-1 (PD-1):PD-L1 negative costimulation is essential for prolonged survival of intratesticular islet allografts, as blocking PD-L1 or PD-1, but not PD-L2 and cytotoxic T-lymphocyte antigen 4, abrogated long-term survival of intratesticular islet allografts. As controls, blocking PD-1 or PD-L1 did not significantly accelerate the acute rejection of islet allografts transplanted under the renal capsule, a conventional islet-grafting site. We also found for the first time that PD-L1 is constitutively expressed mainly by spermatocytes and spermatids in seminiferous tubules of the testis. Moreover, infiltrating T cells underwent less vigorous proliferation but faster apoptosis in the testis than in the kidney. Blocking PD 1:PD-L1 costimulation largely abolished the suppression of T-cell proliferation and acceleration of T-cell apoptosis. Importantly, testicular immune privilege significantly suppressed the generation and proliferation of donor-specific memory CD8 T cells. CONCLUSIONS: The constitutive expression of PD-L1 in the testis is an important mechanism underlying testicular immune privilege and long term survival of intratesticular islet allografts. PMID- 19543054 TI - Prolonged survival of fully mismatched cardiac allografts and generation of regulatory cells by Sairei-to, a Japanese herbal medicine. AB - Sairei-to (TJ114), a 12-component Japanese herbal medicine, is used to treat immune-related diseases. We investigated the effects of oral administration of TJ114 in a murine model of cardiac transplantation with fully mismatched allografts. Untreated CBA mice rejected C57BL/6 hearts acutely (median survival time [MST], 7 days), whereas survival of allografts from mice given TJ114 was significantly prolonged (MST >100 days). Secondary CBA recipients of C57BL/6 hearts also had prolonged allograft survival (MST >100 days) after adoptive transfer of whole or CD4 splenocytes from primary CBA allograft recipients given TJ114. None of the individual components of TJ114 prolonged allograft survival, suggesting that its effects require administration of the combination agent. In mixed leukocyte cultures, proliferation of splenocytes from TJ114-treated CBA recipients was markedly suppressed compared with that of splenocytes from untreated mice, and interferon-gamma production was significantly reduced. Thus, in our model, TJ114 treatment induced hyporesponsiveness to cardiac allografts and generated CD4 regulatory cells. PMID- 19543055 TI - Human CMV infection of porcine endothelial cells increases adhesion receptor expression and human leukocyte recruitment. AB - BACKGROUND: Potential xenozoonosis is a concern for the clinical application of xenotransplantation. Human cytomegalovirus (HCMV) is one of the most important pathogens in allotransplantation, but the consequences of HCMV cross-species infection of porcine xenografts are unknown. Therefore, we investigated the effects of HCMV infection of porcine endothelial cells (pEC) on cell surface molecule expression and human leukocyte recruitment. METHODS: Infection of pEC inoculated with untreated, UV-inactivated, or heparin-treated HCMV at a multiplicity of infection (MOI) of 1 was analyzed by immediate early (IE) antigen expression. Cell surface receptor expression was studied by flow cytometry on pEC bulk cultures and differentially on IE-positive and -negative pEC. Adhesion of human leukocytes was tested on pEC monolayers. pEC supernatants were analyzed for cytokine content, chemotactic activity, and stimulatory effect on resting secondary pEC cultures. RESULTS: At day 2 postinfection, IE staining was evident in 10% to 20% of HCMV-infected cells. Cell-surface expression of E-selectin and vascular cell adhesion molecule-1 (VCAM-1) was upregulated in both IE-negative and -positive fractions of HCMV-infected pEC. In contrast, porcine major histocompatibility complex class I expression was upregulated in IE-negative cells, but reduced in IE-positive cells. The receptor alterations in the IE negative fraction were mediated by pEC-derived soluble factors. The increased adhesion receptor expression was paralleled by enhanced human leukocyte chemotaxis and adhesion to infected pEC cultures. Pretreatment of HCMV with heparin, but not UV-inactivation, prevented adhesion-receptor modulation and reversed the increased adhesion and chemotaxis. CONCLUSIONS: After pig-to-human solid organ transplantation HCMV may infect and activate the porcine endothelium, rendering the xenograft more susceptible to human leukocyte recruitment and rejection. PMID- 19543056 TI - Genotypic variation and phenotypic characterization of granzyme B gene polymorphisms. AB - BACKGROUND: Granzyme B has been associated with allograft rejection in solid organ transplantation. Single nucleotide polymorphisms (SNPs) in the granzyme B gene might impact its expression. The aims of this study were (1) to establish the frequency of two granzyme B SNPs (A-295G; Q-55R) in pediatric heart transplant (PHTx) recipients and (2) to determine their phenotypic expression in healthy individuals. METHODS: Three hundred ninety-six PHTx patients (245 white non-Hispanic, 49 black non-Hispanic, 82 Hispanics, and 20 others) and 52 healthy controls were screened for Q-55R and A-295G. For the control samples, we assessed the frequency of granzyme B positive cells by ELISPOT assay after mitogen stimulation. RESULTS: Among the PHTx recipients, 57% percent of the population carried the Q/Q genotype, whereas 6% were R/R homozygotes. Seven of 49 (14%) black non-Hispanics were R/R homozygotes, whereas 13 of 245 (5%) of white non Hispanics and 5 of 82 (6%) Hispanics carried the R/R genotype (P=0.02). The A allele frequency of granzyme B A-295G (49.6%) was similar to that of the G allele (50.4%). However, 80% of Black non-Hispanics were A allele carriers compared with 68% of White non-Hispanics (P<0.0001). After mitogen stimulation, the frequency of granzyme B positive cells was higher in the Q/Q homozygotes compared with R/R carriers (P=0.006), whereas a similar frequency of granzyme B positive cells was noticed among the genotypes of A-295G SNP. CONCLUSIONS: These data indicate that 55 Q/Q genotype is associated with increased in vitro expression of granzyme B. PMID- 19543057 TI - Urinary Peptide patterns in native kidneys and kidney allografts. AB - BACKGROUND: The use of biopsies to determine kidney health after kidney transplantation is an invasive procedure with some risk to the patient. Consequently, a noninvasive test for transplanted kidney health would provide a significant advantage over current clinical practice. METHODS: Urines from kidney donors before nephrectomy, pretransplant patients with native kidney disease, and posttransplant kidney recipients were examined for protein biomarkers to diagnose or prognose kidney disease. Proteins were extracted by C4 reverse phase affinity and analyzed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: Urine from individuals with healthy kidneys showed few components other than two ubiquitous saposin B glycoisoforms. Patients with kidney disease lacked saposin B and showed new components in two patterns: the most common contained beta-2 microglobulin (B2M, m/z=11,732) plus one or more peaks at m/z=10,350, 9480, 4337, and 4180. Pattern 2 lacked beta-2 microglobulin but contained several degradation products of alpha-1 antitrypsin. Other pathologic components included urinary protein 1 (m/z=15,835), transthyretin (m/z=13,880), and a component at m/z=13,350. CONCLUSIONS: Patients with acute rejection showed profiles that ranged from those of kidney donors to those of advanced kidney disease. The range of patterns may be useful for analysis of transplant patients without complications and persons with developing kidney disease before or after transplant. PMID- 19543058 TI - Noninvasive detection of acute and chronic injuries in human renal transplant by elevation of multiple cytokines/chemokines in urine. AB - BACKGROUND: Injuries in kidney transplant is currently diagnosed by needle biopsy. A noninvasive test that sensitively detects these injuries would benefit the patients. METHODS: Urine samples were collected from healthy controls and kidney transplant recipients. Urine samples were screened first with an antibody array consisting of 120 chemokines and cytokines and then with a multiplex beads assay. Representative parameters, including macrophage inflammatory protein 1Delta, osteoprotegerin, monokine induced by interferon-gamma (IFN), and IFN gamma-induced protein of 10 kDa, were simultaneously determined by a quadruplex assay in urine samples from 84 patients with renal allograft injury, 29 patients with stable graft function, and 19 healthy individuals. RESULTS: Twenty-three cytokines/chemokines were found to be elevated in urine samples of patients with acute rejection by the antibody array. The second round of screening confirmed that 11 of the 23 parameters were elevated in the patients but not in the healthy controls. Induced protein of 10 kDa and monokine induced by IFN-gamma were significantly elevated in urine samples of patients with acute renal injury, and macrophage inflammatory protein-1Delta and osteoprotegerin were significantly elevated in patients with both acute and chronic renal injuries. The combination of the four parameters had a high positive detection rate (97.6%) for renal transplant injury and could differentiate between acute and chronic injury. CONCLUSION: These results might indicate that the present multiplex assay provides a basis to establish a noninvasive method for the diagnosis and monitoring of renal transplant injury. PMID- 19543059 TI - Calcineurin inhibitor-based immunosuppressive therapy, donor age, and long-term outcome after kidney transplantation. AB - BACKGROUND: It is unclear whether the choice of maintenance immunosuppression modulates the negative effect of advanced donor age on outcome after renal transplantation. METHODS: All 1829 patients who received their first transplant between 1990 and 2003 at the Vienna Medical Centre and had a functioning graft after 90 days were studied. At this time point, 1587 received calcineurin inhibitors (CNI+), 242 did not (CNI-). Actual and functional graft survival was analyzed in subgroups based on donor age (<36, 36-49, 50-64, and >64 years) and immunosuppressive therapy. RESULTS: The median follow-up time was 7 years. In total, we observed 312 deaths and 275 graft losses. After adjusting for several variables considered as potential confounders, actual graft survival was better in CNI+ patients compared with CNI- patients only if donor age was less than 36 years (adjusted hazard ratio 0.25, 95% confidence interval 0.17-0.38) or 36 to 49 years (0.43, 95% confidence interval 0.29-0.62). Similar results were obtained for functional graft survival. Patient survival was significantly better in CNI+ subjects irrespective of donor age (0.41, 95% confidence interval 0.30-0.57). DISCUSSION: Use of CNI 90 days after transplantation is associated with improved patient survival even after adjustment for confounders, but its beneficial association with actual and functional graft survival is lost or at least reduced if kidneys from donors older than 50 years are used. PMID- 19543060 TI - Renal transplant from very old donors: how far can we go? AB - BACKGROUND: The organ shortage has led many transplant centers to accept kidneys from old, suboptimal deceased donors, and make increasing use of old-for-old allocation systems. We report the experience of an Italian transplant center in the utilization of "ultra-old" (>75 years old) donors. METHODS: Sixty grafts from donors aged 75 years or older (mean age 79.1 years, range 75-90 years) were used for 38 patients: 16 as single and 22 as double transplants. RESULTS: The actuarial graft survival rate was 73.7% for year 1, 69.8% for year 2, and 64.0% for year 3. The patient survival rate was 81.2% and remained stable for years 1, 2, and 3. The delayed graft function rate was 57.9%. Acute rejection and chronic allograft nephropathy rates were comparable with our other expanded criteria donors. The majority of the patients had stable creatinine levels, between 2 and 3 mg/mL after the second month, with sufficient creatinine clearance. CONCLUSIONS: Our results seems encouraging with patient and graft survival rates, complication rates, and renal function parameters being slightly worse than in expanded criteria donors, but still generally acceptable. The use of old kidneys in old recipients, bearing in mind their usual life expectancy, gives them a properly functioning kidney and improved quality of life. PMID- 19543061 TI - Rituximab and intravenous immunoglobulin treatment of chronic antibody-mediated kidney allograft rejection. AB - Kidney transplant rejections are classified into T-cell-mediated and antibody mediated rejections (AMR). C4d staining on allograft biopsies and solid-phase assays to measure donor-specific alloantibodies have helped to precisely define the latter. Although for acute AMRs, therapy mainly relies on plasmapheresis or immunoadsorption, no studies for treatment of chronic AMR are available. Here, we report on four kidney allograft recipients suffering from chronic AMR 1 to 27 years posttransplant, who were treated with a combination of rituximab and intravenous immunoglobulin (IVIG). Rituximab/IVIG improved kidney allograft function in all four patients, whereas donor-specific antibodies were reduced in 2 of 4 patients. However, in one patient an acute rejection episode occurred 12 months after this treatment, and another patient had severe, possibly rituximab associated lung toxicity. Thus, rituximab/IVIG may be a useful strategy for the treatment of chronic AMR, but further randomized multicenter studies are necessary to establish its efficacy and safety profile. PMID- 19543062 TI - Hepaticoduodenostomy is an alternative to Roux-en-Y hepaticojejunostomy for biliary reconstruction in live donor liver transplantation. AB - INTRODUCTION: A Roux-en-Y hepaticojejunostomy (HJ) is usually performed during live donor liver transplantation (LDLT) when a duct-to-duct reconstruction is not possible. However, direct anastomosis of the bile duct to the duodenum (hepaticoduodenostomy [HD]) is an alternative technique for biliary repair that has been previously used for conventional biliary surgery and at our center for cadaveric liver transplant. We provide the first evidence that HD is an alternative technique for biliary reconstruction in LDLT. METHODS: We performed a total of 71 LDLT between 2002 and 2008. An end-to-end anastomosis was used in 30 patients. Forty-one patients had a biliary enteric anastomosis in which seven were reconstructed with an HD. Accessory ducts were fashioned into a common duct or implanted into the duodenum separately. RESULTS: There were no patient deaths or retransplants in a follow-up period that ranged from 90 to 771 days after surgery. One patient was diagnosed with cholangitis that responded to intravenous antibiotics and removal of the stent by endoscopy. CONCLUSIONS: This preliminary case series suggests that that HD is a feasible alternative to HJ biliary anastomosis when a duct-to-duct anastomosis cannot be performed. HD offers the possible advantage of simple postoperative access to the biliary system by endoscopy and avoids complications caused by HJ bowel anastomosis. PMID- 19543063 TI - Risk of fractures after renal transplantation in the United States. AB - BACKGROUND: Although it is known that the incidence of fracture events is increased in renal transplantation recipients, the timing and the factors associated with risk of fractures are less well understood. The objective of this study was to estimate the time to fracture in renal transplantation recipients and to determine whether risk was associated with patient and transplantation characteristics. METHODS: Using the U. S. Renal Data System, we retrospectively studied 68,814 patients, who underwent renal transplantation between 1988 and 1998. Fractures were identified from International Classification of Diseases, Ninth Revision, Clinical Modification diagnosis codes in billing data. Time to first fracture was modeled during the first 5 years posttransplant using the Kaplan-Meier method and Cox proportional hazards models. RESULTS: Of the patients who underwent transplantation, 22.5% developed a fracture within 5 years. Woman (hazard ratio [HR] 1.36, P<0.0001), patients older than 45 years of age (HR 1.14, P<0.0001) especially older than 65 years (HR 1.69, P<0.0001), and whites (HR 1.28, P<0.0001) were at increased risk of a fracture. Additionally, receipt of a deceased donor kidney (HR 1.30, P<0.0001), increased human leukocyte antigen mismatches (HR 1.09, P<0.014), diabetes (HR 1.88, P<0.0001), pretransplant dialysis (HR 1.08, P<0.0001), and an aggressive induction immunosuppression regimen (HR 1.14, P<0.0001) all significantly increased risk of fracture events during the first 5 years. CONCLUSIONS: In addition to patient demographic features, donor factors, including suboptimal organ quality and the need for more intense immunosuppression, were associated with an increased risk of fractures during the first 5 years after a renal transplant. PMID- 19543064 TI - Experience with immune monitoring in lung transplant recipients: correlation of low immune function with infection. AB - BACKGROUND: Lung transplants, in particular, have the highest rate of infections among solid organ transplant recipients. However, there is no existing objective measure to predict the development of infections. We report the correlation between Cylex ImmuKnow (ng/mL ATP) values and various infectious syndromes in a large prospective cohort of lung transplant recipients. METHODS: We followed up 175 lung transplants that developed 129 infectious episodes. Multiple logistic regression analysis was performed; generalized estimating equations were used to determine the odds ratio for infections. RESULTS: The median ImmuKnow values in cytomegalovirus disease (49.3 ng/mL ATP), viral infection (70 ng/mL ATP), and bacterial pneumonia (92 ng/mL ATP) were significantly different from stable state (174.8 ng/mL ATP). The median ImmuKnow values of fungal disease (85 ng/mL ATP) and tracheobronchitis (123 ng/mL ATP) had a tendency to be lower than stable state (P=0.10), whereas patients with fungal colonization had comparable ImmunKnow values (167 vs. 174.8 ng/mL ATP). Of the patients colonized with fungus who subsequently developed fungal disease within 100 days, the median value of ImmuKnow was significantly lower than in those who did not develop fungal disease (22.5 vs. 183.5 ng/mL ATP; P<0.0001). Generalized estimating equation regression analysis showed ImmuKnow values less than or equal to 100 ng/mL ATP to be an independent predictor of infections (odds ratio 2.81). CONCLUSIONS: Cylex ImmuKnow assay monitoring has the potential to identify the patients at risk of developing infection and those colonized with fungus that are at risk of developing disease. PMID- 19543065 TI - Donor risk index and MELD interactions in predicting long-term graft survival: a single-centre experience. AB - INTRODUCTION: Feng et al. described the donor risk index (DRI) in North American liver transplant recipients. We evaluated the effect of the DRI and model for end stage liver disease (MELD) score on liver transplant recipients from a single center in the United Kingdom. METHOD: Prospectively, collected data of all patients transplanted at our center between January 1995 and December 2005 were included in the analysis (n=1090). Outcomes evaluated included patient-censored and death-censored graft survival. Outcomes of liver transplantation from "high" and "low" DRI groups (> or =1.8 and <1.8, respectively) on patients categorized into low (<15), intermediate (15-30), and high (>30) MELD categories were analyzed. RESULTS: MELD at transplant was the only significant predictor of patient survival. MELD at transplant and DRI more than 1.7 were associated with a poorer graft survival (P=0.03). There was a trend toward poorer graft survival in high DRI grafts transplanted in low and "intermediate" MELD categories (P=0.47 and 0.006, respectively). However, in the high MELD category, there was a similar graft survival for both high and low DRI grafts. CONCLUSION: Patients with low and intermediate MELDs at transplantation may be better served by a low DRI graft, whereas patients with high MELD may not be compromised by receiving a high DRI graft. PMID- 19543066 TI - Association of arterial hypertension with renal target organ damage in kidney transplant recipients: the predictive role of ambulatory blood pressure monitoring. AB - BACKGROUND: Although arterial hypertension is a powerful predictor of graft failure, only few studies have evaluated 24-hr blood pressure (BP) profile in renal transplant recipients (RTRs). METHODS: We performed ambulatory blood pressure monitoring (ABPM) in 94 RTRs (65 men; age 28-71 years) with 1-year functioning grafts. Serum biochemical parameters, daily proteinuria, and transplantation-related data were evaluated in all subjects. RESULTS: ABPM showed that only 5% of RTRs were normotensives (BP<130/80 mm Hg) and identified 29% of patients with nocturnal hypertension. A strong, direct correlation was shown between each set of both systolic BP and diastolic BP measured by ABPM and serum creatinine, daily proteinuria, and serum triglycerides (P at least <0.025 for each). Serum creatinine immediately after transplantation and 1-yr asleep diastolic BP were the only significant predictors of 1-yr creatinine (P<0.0001; r=0.49), whereas awake systolic BP was the only predictor of daily proteinuria (r=0.39; P=0.005) by multiple regression analysis. CONCLUSIONS: BP assessed by ABPM proved to be a stronger predictor of renal graft damage than traditional immunologic factors. ABPM improved the diagnostic accuracy of arterial hypertension in RTRs and was the only effective tool in disclosing the association of BP with 1-year renal transplant outcome. PMID- 19543068 TI - Cyclosporine withdrawal improves long-term graft survival in renal transplantation. AB - BACKGROUND: The reduction in renal transplant rejection rates achieved over the last 20 years have not translated into a commensurate improvement in long-term graft survival. Cyclosporine has been central to immunosuppressive regimens throughout this period but its effect on long-term transplant outcomes remains unclear. METHODS: This randomized controlled trial allocated first cadaveric renal transplant recipients in seven centers around Australia to three immunosuppressive regimens: azathioprine and prednisolone (AP), long-term cyclosporine alone (Cy), or cyclosporine initiation followed by withdrawal at 3 months and azathioprine and prednisolone replacement (WDL). RESULTS: Between 1983 and 1986, 489 patients were randomized with 98% follow-up to a median of 20.6 years. Mean graft survival (censoring deaths) was superior in the WDL group (14.8 years) when compared with both AP (12.4 years, P=0.01 log-rank test) and Cy (12.5 years, P=0.01 log-rank test) groups by intention-to-treat. Without death censoring, graft survival with WDL was superior to AP (9.5 years vs. 6.7 years, P=0.04) and of borderline superiority to Cy (9.5 years vs. 8.5 years, P=0.06). Patient survival was not different between the three groups. Renal function was superior in AP (at 1, 10, and 15 years posttransplant) and WDL (at 1, 5, 10, 15, and 20 years) groups when compared with Cy. CONCLUSION: This study illustrates superior long-term renal transplant survival and preservation of renal function with a protocol using cyclosporine withdrawal. If long-term renal transplant outcomes are to improve, we should reconsider guidelines recommending universal maintenance use of cyclosporine. PMID- 19543067 TI - Insulin hyposecretion in nondiabetic, tacrolimus-treated renal transplant recipients more than 6 months posttransplantation. AB - BACKGROUND: New-onset diabetes after transplantation is an important complication of renal transplantation. Few studies have examined nondiabetic renal transplant recipients for occult defects in insulin sensitivity or secretion. The aims of this study were to identify abnormalities of glucose metabolism in nondiabetic, tacrolimus-treated renal transplant recipients more than 6 months posttransplantation and characterize determinants. METHODS: Eighteen nondiabetic renal transplant recipients and 20 healthy control subjects were examined with a frequently sampled intravenous glucose tolerance test and meal tolerance test to derive first-phase insulin secretion (FPIR), insulin sensitivity, and second phase insulin secretion. RESULTS: FPIR was higher in controls compared with transplant recipients (2225 vs. 1173 pmol/L; P=0.003). In transplant subjects, there was a strong positive linear correlation between glomerular filtration rate (GFR) and FPIR (Pearson correlation r=0.7; P=0.002). There were strong negative correlations between trough tacrolimus concentration at study entry (r=-0.56; P=0.01) and 3-month tacrolimus exposure (r=-0.49; P=0.02) with GFR, but not with FPIR. CONCLUSIONS: Nondiabetic renal transplant recipients had lower FPIR than controls with normal renal function. Declining FPIR predicts type 2 diabetes in the general population. Interpreting the relationship between FPIR and GFR requires exploration of dual tacrolimus toxicity on both renal and pancreatic beta-islet cell function. PMID- 19543069 TI - Use of the abdominal rectus fascia as a nonvascularized allograft for abdominal wall closure after liver, intestinal, and multivisceral transplantation. AB - INTRODUCTION: Abdominal wall closure management has become an important challenge during recipient candidate selection, at the time of donor to recipient matching and during the planning of the surgical procedure for intestinal or multiorgan transplantation. Different strategies have been proposed to overcome the lack of abdominal domain: to reduce the graft size or to increase the abdominal domain. Based on the recent concept of using an acellular dermis matrix (Alloderm) and the availability of abdominal wall tissues from the same organ donor, we conceived the idea of using the fascia of the rectus muscle (FoRM) as a nonvascularized tissue allograft. MATERIALS AND METHODS: This is a retrospective report of a series of 16 recipients of FoRM as part of a liver, intestinal, or multiorgan transplant procedure performed between October 2004 and May 2008 at three different transplant centers. RESULTS: Of the 16 recipients of FoRM, all but one case was performed during their transplantation (four multivisceral, two modified multivisceral, three isolated intestine, and two livers). Five patients underwent a retransplant surgery (two livers, two multivisceral, and one isolated intestine). Abdominal wall infection was present in 7 of 16 cases. Nine patients are still alive. No deaths were related to wound infection. Long-term survival showed complete wound healing and only one ventral hernia. DISCUSSION: The use of a nonvascularized FoRM is a novel and simple surgical option to resolve complex abdominal wall defects in liver/intestinal/multivisceral transplant recipients when it can be covered with the recipient skin. PMID- 19543071 TI - Are all donor-specific antibodies detected by solid-phase assay before transplantation clinically relevant? PMID- 19543073 TI - Mesenchymal stem cell and immunosuppressive drug interactions. PMID- 19543072 TI - Pyoderma vegetans in a renal transplant recipient: first case of human infection with Nocardia vinacea. PMID- 19543070 TI - Comparative outcome analysis of ABO-incompatible and positive crossmatch renal transplantation: a single-center experience. AB - BACKGROUND: ABO-incompatible (ABOi) and positive crossmatch (XM) renal transplants pose special immunologic challenges. It is important to compare outcomes, study resource utilization, and attempt to risk stratify patients in these higher risk transplant settings. METHODS: We compared apheresis utilization and transplant outcomes in ABOi, XM, and combined ABOi-XM renal transplants. We also analyzed multiple parameters, including patient and laboratory variables, to identify predictors of transplant outcome. RESULTS: Incidences of early (< or =30 days posttransplant) antibody-mediated rejection (AMR) and acute cellular rejection (ACR) were similar among the three incompatible groups whereas they differed in allograft rejection for late (>30 days posttransplant) AMR and ACR. Notably, there were no episodes of late AMR among ABOi patients. Patients treated with more than four pretransplant plasmapheresis/intravenous immunoglobulin (PP/IVIg) had a greater likelihood of experiencing early AMR. The median number of posttransplant PP/IVIg treatments was greater than twofold higher in ABOi-XM and XM patients compared to ABOi patients. Patients who required more than five posttransplant PP/IVIg procedures and those with one or more prior renal transplants had higher incidences of late ACR. CONCLUSIONS: Our analysis aids in defining apheresis resource utilization and helps in risk stratification of incompatible renal transplantation. It also aids in predicting allograft rejection and provides an opportunity for preemptive monitoring and treatment. PMID- 19543075 TI - Belatacept as immunosuppression in patient with recurrence of hemolytic uremic syndrome after renal transplantation. PMID- 19543076 TI - Feasibility and efficacy of preoperative chemotherapy with docetaxel, cisplatin and S-1 in gastric cancer patients with para-aortic lymph node metastases. AB - We performed preoperative chemotherapy with combined docetaxel, cisplatin and S-1 (DCS therapy) for treatment of advanced gastric cancer with para-aortic lymph node metastases. The aim of this study was to determine the maximum tolerated dose (MTD) and the dose-limiting toxicities. Furthermore, we evaluated the feasibility of DCS therapy in a preoperative setting, and also examined the pathological response. Fifteen patients received intravenous docetaxel and cisplatin (30, 35 or 40 mg/m2, each dose escalation was reciprocal) on days 1 and 15 and oral S-1 (40 mg/m2 twice daily) on days 1-14 every 4 weeks. After one cycle of chemotherapy, toxicities were evaluated and after two cycles of chemotherapy, patients who were judged to be candidates for curative resection underwent gastrectomy with D2 lymphadenectomy plus para-aortic lymph node dissection. The MTD of this combination was presumed to be at dose level 3 (docetaxel 40 mg/m2 and cisplatin 35 mg/m2). The dose-limiting toxicities were grade 4 neutropenia in one patient, grade 3 febrile neutropenia in two patients and grade 3 diarrhoea in two patients. Thirteen of the 15 patients received complete resection and there was no operation-related death. Good pathological responses were observed in 12 cases with lesions in the lymph nodes (complete response, n = 4; partial response, n = 8) and 11 patients with primary stomach lesions (complete response, n = 2; partial response, n = 9). This preoperative DCS therapy was considered feasible and provided a high pathological response rate in gastric cancer patients with para-aortic lymph node metastases. PMID- 19543077 TI - Relationship between chromogenic factor X and international normalized ratio differs during early warfarin initiation compared with chronic warfarin administration. AB - Chromogenic factor X (CFX) monitoring is necessary in patients with potential international normalized ratio (INR) artifacts during warfarin therapy. The relationship of CFX with the INR needs to be quantitated to have warfarin protocols that are equivalent with either test as a monitoring parameter. This study investigated whether the CFX/INR relationship is different during warfarin initiation compared with that during chronic warfarin therapy. Outpatients (N = 164) taking chronic doses of warfarin and inpatients (N = 137) initiating warfarin therapy had plasma samples tested for CFX and INR. The best fit mathematical relationship of CFX and INR was determined for both groups. A six hundred and twenty-five bed, adult-only, private, tertiary care teaching hospital was the setting of the study. The best fit equation for chronic warfarin patients was quadratic using a reciprocal transformation of the INR. The best fit equation for the warfarin initiation patients was linear using logarithmic transformation of CFX and INR. The predicted CFX from INRs over the range of 1.4-2.2 was 7-18% higher in the warfarin initiation patients than in the chronic warfarin patients. Translation of CFX values into equivalent INRs for use in warfarin initiation and maintenance protocols is improved when using equations specific to the patient situation. PMID- 19543078 TI - Challenges of defining reliable clinical surrogate end points in haemophilia trials: a critical review. AB - The success of a treatment in haemophilia patients experiencing a bleeding episode is very difficult to define. A variety of efficacy assessment tools have been developed in an effort to better assess when haemostasis has been achieved. These assessment tools are particularly important for the evaluation of the efficacy of therapeutic agents whose mechanism of action is based on pharmacological activity in haemostasis rather than upon the principle of 'replacement therapy'. This review focuses on a number of efficacy measures, summarizing their methodology and discussing their validity. In addition, future developments and requirements in order to evaluate the effectiveness of haemostatic treatment are discussed. The majority of end points used for evaluation of haemostasis relate to the relief of symptoms arising from bleeds. The results of this review highlight that several efficacy end points are frequently combined in order to provide a more comprehensive assessment of efficacy. Key limitations of current methodology are the subjectivity of assessment by either the patient or clinician, and the incomparability of results between trials. PMID- 19543079 TI - Testing for platelet function: do we diagnose inherited disorders appropriately? PMID- 19543081 TI - Resistance training reduces the blood pressure response of older men during submaximum aerobic exercise. AB - OBJECTIVE: The objective of this study was to determine whether 16 weeks of resistance training (RT) can reduce the blood pressure response and improve the cardiovascular function of men aged 70-80 years during submaximum aerobic exercise. METHODS: Twenty-four men aged between 70 and 80 years were randomly assigned to an RT group (n = 12) and control group (n = 12). Training consisted of three sets of six to 10 repetitions at 70-90% of one repetition maximum, three times per week, on an incline squat machine for 16 weeks. Blood pressure and cardiovascular function were assessed during submaximum cycle exercise at 40 W, and 50 and 70% of maximum oxygen consumption (VO2max) before training and after 16 weeks of training. Leg strength and VO2max were assessed every 4 weeks of the 16-week study. RESULTS: At 40 W, heart rate, systolic blood pressure, and rate pressure product were lower and stroke volume was significantly higher after 16 weeks of training. At 50% VO2max, heart rate and rate pressure product were lower after 16 weeks of training and at 70% VO2max, cycle ergometry power, VO2, and arterio-venous oxygen difference were higher after 16 weeks of training. Leg strength significantly increased after 16 weeks of training. CONCLUSION: Sixteen weeks of RT significantly reduces the blood pressure response and improves the cardiovascular function of older men during submaximum aerobic exercise. Therefore, RT not only increases muscular strength and hypertrophy but also provides significant cardiovascular benefits for older individuals. PMID- 19543080 TI - Reduction of morning blood pressure surge after treatment with nifedipine GITS at bedtime, but not upon awakening, in essential hypertension. AB - OBJECTIVES: The extent of morning blood pressure (BP) surge upon wakening has been associated with increased incidence of stroke and cardiovascular mortality. This trial investigated the antihypertensive efficacy and effects on the morning BP surge of awakening versus bedtime administration of nifedipine in essential hypertension. METHODS: We studied 238 previously untreated hypertensive patients (108 men and 130 women), 53.3+/-11.4 years of age, randomly assigned to receive nifedipine (30 mg/day) as a monotherapy either upon awakening or at bedtime. BP was measured for 48 h before and after 8 weeks of treatment. RESULTS: The BP reduction after the treatment was significantly greater with bedtime dosing (P<0.001). The proportion of patients with controlled ambulatory BP thus increased from 28 to 43% (P = 0.019) with bedtime treatment. The sleep time relative BP decline was unchanged after morning treatment, but increased toward a more dipping pattern after bedtime dosing (P = 0.026 between groups). The morning BP surge was unchanged after the administration of nifedipine upon awakening (1.4/1.2 mmHg reduction in systolic/diastolic BP surge, P>0.270), but significantly reduced after bedtime dosing (6.2/4.4 mmHg reduction, P<0.001). CONCLUSION: Nifedipine efficiently reduces BP for the entire 24 h and to a significantly larger extent after bedtime administration. The significant added efficacy on reducing night-time BP, the decrease in the prevalence of a nondipper BP pattern, and the significant decrease in morning BP surge (all relevant markers of cardiovascular morbidity and mortality) of bedtime as compared with morning administration, consistently indicate that nifedipine should preferably be administered at bedtime in patients with essential hypertension. PMID- 19543082 TI - Childhood-onset hemiatrophy caused by unilateral morphea. PMID- 19543083 TI - The myeloperoxidase gene and its influence on myocardial infarction in a Swedish population: protective role of the -129A allele in women. AB - OBJECTIVE: The aim of this study is to investigate the potential role of functional myeloperoxidase (MPO) promoter polymorphisms in the occurrence of myocardial infarction (MI) in the Stockholm Heart Epidemiology Program. METHODS AND RESULTS: Two MPO promoter polymorphisms, -129G/A and -463G/A, were genotyped in the Stockholm Heart Epidemiology Program population (n = 2774). The -129A allele was associated with a lower risk of MI in women [odds ratio (OR) (95% confidence interval): 0.65 (0.43-0.98), P = 0.03] but not in men [OR: 1.12 (0.86 1.47), P = 0.38]. When women were stratified by age and hormone replacement therapy, the protective effect of the -129A allele was only evident in women younger than 55 years or not receiving hormone replacement therapy. In these two groups, OR (95% confidence interval) for carriers of the -129A allele were 0.34 (0.12-0.92) (P = 0.03) and 0.51 (0.32-0.81) (P = 0.004), respectively. For the 463G/A polymorphism, no associations to MI risk were observed either in women or in men. CONCLUSION: The A allele of the MPO -129G/A promoter polymorphism is associated with a reduced MI risk in women. PMID- 19543085 TI - Duration of treadmill exercise testing combined with QRS score predicts adverse cardiac outcome at long-term follow-up. AB - OBJECTIVE: Total exercise duration and abnormal QRS score values are treadmill exercise testing (TET) prognostic parameters that have been shown to be significantly and independently associated with cardiac mortality. We evaluated the prognostic value of a new index (M score, Michaelides score) incorporating TET duration and QRS score values in a simple index. METHODS: In this study, we included 626 patients, who underwent TET and coronary arteriography. Cardiac catheterization showed the presence of coronary artery disease in 64.3% of these patients. The M score was calculated by adding the value of the Athens QRS score to the duration of TET (in minutes). The outcome measure was a composite of myocardial infarction or death. Patients were prospectively followed for 38+/-21 months (median 36 months). RESULTS: The composite endpoint was more frequent among the patients of the 1st quartile (M-score values <-5.8). In univariate analysis, mortality of the first-quartile patients was significantly higher (14 vs. 1.1%, P<0.001). In multivariate Cox's regression analysis for age, sex, diabetes, smoking status, hypertension, hypercholesterolemia, maximum ST depression at TET, angina during TET, coronary artery disease on angiography, and echocardiographic left ventricular ejection fraction, the first quartile of M score values was found to be independently associated with the composite endpoint (relative risk = 3.26, 95% confidence interval = 2.01-5.29, P<0.001). CONCLUSION: This study shows that a new index termed the M score, which incorporates QRS score and exercise duration, predicts mortality and occurrence of myocardial infarction at long-term follow-up of high-risk individuals, independently of TET induced ST-segment changes. PMID- 19543084 TI - Safety and efficacy of a prolonged bivalirudin infusion after urgent and complex percutaneous coronary interventions: a descriptive study. AB - OBJECTIVE: Bivalirudin, a direct thrombin inhibitor, provides similar ischemic outcomes with significantly less major bleeding compared with unfractionated heparin (UFH) plus a glycoprotein IIb/IIIa inhibitor (GPI) in patients undergoing percutaneous coronary interventions (PCI). Although the approved labeling for bivalirudin allows for low-dose prolonged postprocedure administration, this practice is not routine. Therefore, we sought to evaluate the safety and efficacy of longer post-PCI infusion. METHODS: From our database, we retrospectively compared two groups of patients with acute coronary syndrome undergoing complex PCI, one group treated with UFH+GPI (n = 59) and another with a periprocedural and post-PCI bivalirudin infusion for 4 h (n = 50). Endpoints included periprocedural myocardial infarction (MI), 30-day major adverse cardiac events, and in-hospital major and minor bleeding. RESULTS: There were no significant differences in the baseline and procedural characteristics of the two groups; most patients (approximately 90%) had complex coronary lesions (the American College of Cardiology/American Heart Association type B2/C). There was no significant difference in the rates of periprocedural MI (11.9 vs. 8.0%, P = NS) or 30-day major adverse cardiac events (8.5 vs. 6.0%, P = NS) among patients treated with UFH+GPI or bivalirudin. However, patients who received bivalirudin had significantly lower rates of minor bleeding (20.3 vs. 4.0%, P<0.05), and a trend toward significantly less major bleeding (8.5 vs. 4.0%, P = 0.07). When we compared the group treated with a prolonged bivalirudin infusion with a historical group treated with peri-PCI-only bivalirudin infusion, we observed in the latter an increased incidence of periprocedural MI and a comparable incidence of bleeding. CONCLUSION: A prolonged bivalirudin infusion after urgent PCI seems effective in protecting myocardium without increasing bleeding rates, and represents an attractive alternative to the standard pharmacological treatment of UFH+GPI in the catheterization laboratory. PMID- 19543086 TI - Spectrum of electrocardiographic and angiographic coronary artery disease findings in patients with cocaine-associated myocardial infarction. AB - BACKGROUND: Cocaine is the most common abused drug in patients presenting to the emergency room with chest pain and frequently leads to cardiac catheterization procedure. The extent of severity underlying coronary artery disease (CAD) in this subgroup of patients has not been well defined. This study set out to define the coronary anatomy as well as the extent of CAD in patients with cocaine associated myocardial infarction (MI) and correlate that to the presenting electrocardiogram (ECG). METHODS: Ninety-seven consecutive patients with documented MI and positive urine drug screen for cocaine metabolites were included in the study. Demographic, clinical, ECG and coronary angiography variables were collected. RESULTS: ST elevation MI was encountered in 32% of the patients. Other ECG findings included ST segment depression, T-wave inversion, left ventricular hypertrophy, conduction blocks and/or old MI in more than 80% of cases. Of the total of 66 patients who underwent angiography, 82% had obstructive CAD, with single-vessel disease being the most frequent finding. None of these presenting ECG findings correlated with angiographic location or severity of obstructive CAD. In nearly one-fifth of the patients, troponin elevation suggestive of cardiac myonecrosis occurred in the absence of ECG findings or angiographic coronary disease. CONCLUSION: The majority of patients with cocaine associated MI have obstructive CAD with predominant single-vessel disease. Although ECG abnormalities are frequently encountered, they are of limited diagnostic value in the clinical decision making. PMID- 19543087 TI - Coronary plaque dimensions and composition by intravascular ultrasound radio frequency lesion segment analysis in stable and unstable angina patients. AB - AIMS: We hypothesized that the plaque composition and plaque type classification differs between acute coronary syndrome (ACS) and stable angina (SA) patients. METHODS AND RESULTS: We analyzed culprit lesion (CL) and nonculprit lesion (NCL) of ACS patients compared with target lesion (TL) and nontarget lesion (NTL) of SA patients by intravascular ultrasound radio frequency analysis in 874 lesion segments of 424 patients (ACS: 193 patients/SA: 231 patients). Comparing all lesion segments in ACS and SA patients did not show significant differences in absolute or relative plaque composition. However, necrotic core area was larger in CL versus TL (0.9+/-0.7 vs. 0.7+/-0.5 mm, P=0.005) and all plaque components were significantly higher in CL compared with NCL and TL compared with NTL, respectively. A higher amount of thin cap fibroatheroma lesions (15.2 vs. 5.1%, P<0.0001) was detected in ACS compared with SA patients. Fibrocalcific lesions were lower in ACS patients (3 vs. 10.5%, P<0.0001). CONCLUSION: The differentiation in CL/NCL of ACS and TL/NTL of SA patients revealed significant differences in plaque composition and plaque types when examined by intravascular ultrasound radiofrequency analysis. However, considerable overlap between plaque characteristics exists for ACS and SA patients. PMID- 19543088 TI - Caustic ingestions in children. AB - PURPOSE OF REVIEW: The purpose of this review is to outline the current epidemiology, mechanism of injury, clinical manifestations, management and long term complications of caustic ingestions in pediatric patients. RECENT FINDINGS: Recent data suggest that more than 200 000 exposures to household or industrial cleaning products occur annually in the United States. It is difficult to determine what fraction of these exposures represents caustic ingestions. Caustic ingestions occur most commonly in children less than age 6 years. Because of the accidental nature of the ingestions, the case fatality rate for pediatric patients is significantly less than that of adolescents and adults. Despite laws to limit the concentration of household cleaning products, farm and industrial products and products stored in nonoriginal containers represent a significant source of caustic agents. Endoscopy remains the preferred method of staging injury. In children the absence of symptoms does not predict lack of relevant injury. However, the presence of three or more symptoms is associated with a high likelihood of significant injury. Long-term complications in pediatric patients may be severe and include esophageal cancer. SUMMARY: Caustic ingestions remain a significant cause of pediatric morbidity in the United States and abroad. Endoscopy is the primary method of staging injury following a caustic ingestion. Extent of injury at initial evaluation remains the best predictor of morbidity and mortality in pediatric patients following an accidental caustic ingestion. PMID- 19543089 TI - Are the new moisturizers more effective? AB - PURPOSE OF REVIEW: The authors discuss the role of moisturizers in the daily skin care regimen of healthy skin and in atopic dermatitis, as well as the efficacy of newer products. RECENT FINDINGS: Recent findings have shown that the affected skin of atopic individuals is deficient in ceramides, an integral lipid component of the stratum corneum. Several manufacturers have developed topical products with ceramides in an attempt to replace the missing molecules. These products, though more expensive, have shown moderate efficacy improvements over traditional moisturizers. SUMMARY: Moisturizers are recommended to nearly every atopic patient, and should be part of the daily skin care of these patients, as they have the potential to reduce the quantity of topical steroids necessary to keep disease quiescent. The vast array of choices available to consumers is overwhelming, and most patients would like their physicians to recommend specific products. We attempt to provide a review of the newer products available, with special emphasis on those products that contain ceramides. PMID- 19543090 TI - Intraocular pressure measurement in children under general anaesthesia with sevoflurane. PMID- 19543091 TI - Increase of propofol requirement after repeated administration for experimental anaesthesia. PMID- 19543092 TI - Multicenter randomized controlled trial of a home walking intervention after outpatient cardiac rehabilitation on health-related quality of life in women. AB - BACKGROUND: Poor health-related quality of life (HRQL) has been shown to be predictive of adverse outcomes in cardiac patients. As women with coronary heart disease have been shown to have lower HRQL than men with coronary heart disease, women are at greater risk of a poor clinical outcome. This study tested the effect of a 12-week home walking intervention after completion of outpatient cardiac rehabilitation (OCR) on HRQL and maintenance of physical activity among women. DESIGN: Multicenter two-group randomized trial. METHODS: After completion of OCR, participants were randomly allocated to the intervention or usual care groups. The outcomes were HRQL (assessed using the MacNew Heart Disease HRQL instrument) and self-reported physical activity (assessed using the Stages of Change model of exercise behavior) at 3, 6, and 12 months after OCR. RESULTS: Seventy-two women were randomized to the intervention and 81 to usual care. Attrition was greater in the treatment group (13 vs. 1%). HRQL scores increased relative to the base level in both arms and were significantly higher in the intervention group at 6 months, but not at 3 or 12 months. Maintenance of physical activity declined over time in both groups, however, this decline was significantly reduced among women in the intervention group. CONCLUSION: HRQL improved in both groups, but seemed to increase earlier among women in the intervention group. As maintenance of physical activity was higher among women in the intervention group, this minimal intervention could be used to facilitate women's progression from supervised to independent exercise. PMID- 19543093 TI - A reliable and valid protocol for measuring maximal oxygen uptake in pigs. AB - BACKGROUND: Most knowledge about cellular and molecular adaptation in the heart after exercise training comes from rodent models, and this has substantially improved our knowledge about exercise-induced cardiac adaptations. However, in rodents, the electrophysiological properties of the heart are different from the human heart. Therefore, the need of exercise-training models in larger animal models is obvious. Physiological studies of cardio-respiratory fitness require training regimens that give robust and adequate testing procedures to quantify the outcome. METHODS: We developed a valid and reproducible protocol for measuring maximal oxygen uptake (VO2max) in young pigs. As previous studies have exercised pigs using horizontal treadmills, we determined whether treadmill inclination may influence the level of peak oxygen uptake (VO2peak) achieved, and whether the true VO2max was reached. Eight young pigs were used. Submaximal and VO2peak were tested at five different inclinations from 13 to 30 degrees . RESULTS: At submaximal VO2, there was an excellent test-retest at all treadmill inclinations (r = 0.99, coefficient of variation = 1.8%). The level of VO2peak was dependent upon treadmill inclination and the true VO2max, defined as a levelling-off of VO2 despite increased running speed, was only reached a treadmill inclination of 24 degrees . For VO2peak we only observed a significant test-retest correlation when using 19 and 24 degrees inclination of the treadmill (r = 0.88, coefficient of variation = 9.7%). CONCLUSION: The use of inappropriate treadmill inclination might hide training-induced adaptations if the true VO2max is not reached. This study shows that the present test protocol can be used in future studies of exercise on treadmill, when the aim is to measure submaximal and VO2max in pigs. PMID- 19543094 TI - Genetic variation in IGF-1 and breast cancer risk in Ashkenazi carriers and noncarriers of BRCA1/2 mutations. AB - Earlier studies indicate that high circulating levels of insulin-like growth factor-1 (IGF-1) may be associated with premenopausal breast cancer. We studied variations in the IGF-1 gene and the growth hormone (GH1) gene in relation to risk of breast cancer in 667 Ashkenazi Jewish women (321 cases, 346 controls) from a population-based case-control study in Northern Israel, and a clinical series of 331 founder BRCA mutation carriers (161 affected, 170 unaffected). All participants were tested for six polymorphisms in the IGF-1 gene and one GH1 polymorphism. Logistic regression models were used to estimate odds ratios for haplotype-specific and genotype-specific age-adjusted risks. Two common IGF-1 haplotypes (ATTCAC, GAGTGT) were found, when compared with the most prevalent haplotype ATTCGC (32.5%), to be associated with a decreased risk of breast cancer in premenopausal noncarrier women only. Age-adjusted odds ratios were 0.5 (95% confidence interval: 0.28-0.92) for ATTCAC and 0.46 (95% confidence interval: 0.24-0.89) for GAGTGT. The GH1 polymorphism did not influence the risk of breast cancer in our study population. The IGF-1 gene seems to be associated with breast cancer risk in premenopausal Ashkenazi Jewish women who are not carriers of mutations in BRCA1/2 genes. PMID- 19543095 TI - Are emergency department visits really a teachable moment? Smoking cessation promotion in emergency department. AB - OBJECTIVE: To compare the effectiveness of smoking cessation counseling in the emergency department (ED) versus in outpatient clinics (OCs) setting. METHODS: Over a 3-month period, smokers and recent quitters presenting to ED or OCs were questioned about their smoking habits and desire to quit. They also completed the Fagerstrom Test for Nicotine Dependence (FTND) questionnaire and Prochaska's stages of change (PSC) survey. Standardized 5 min counseling session was carried out, and stop smoking pamphlet and phone number of the hospital's smoking cessation unit were given. One month after initial counseling, patients were telephoned, FTND, PSC, desire to quit, and daily cigarette consumption were asked. Data from those unable to be contacted within 6 weeks were excluded from analysis. RESULTS: Of the 392 patients (197 ED, 195 OC) counseled initially, 340 (87%) were reached for telephone follow-up. Counseling was effective in both groups: FTND and PSC scores had improved, and daily cigarette consumption decreased significantly (17.17-12.49 cigs/day; P=0.000). Smokers counseled in the ED were found more inclined to stop smoking compared with smokers who counseled in OCs, after 1 month of the intervention (95% confidence interval=14.7-7.5%; P=0.051). Only one patient (0.6%) from the ED and 10 (6.6%) from the OC attended the smoking cessation program. CONCLUSION: ED-based counseling for smoking cessation was as effective as that performed in the OC setting. Referral of smokers from the ED to a smoking cessation program was unsuccessful in our patient population. PMID- 19543096 TI - Impact of a program to improve pain management in an emergency department. AB - The objective of this study is to assess the impact of various actions in the management of pain in a pediatric hospital emergency department. This is an observational study, preaction (phase 1) and postaction (phase 2), with two cohorts of patients diagnosed with abdominal pain, chest pain, and severe headache. Between the two phases, various actions were carried out (distribution of pain assessment scales and a new guide for the management of pain, and the holding of clinical training sessions). Three hundred patients were included in the study, with an average age of 9 years and average evolution time of pain of 20 h. Pain assessment in phases 1 and 2 was 30 and 99.3%, respectively. Analgesics were administered to 23% (phase 1) and 38.6% (phase 2) of the patients with pain. No side-effects from the analgesics given were recorded. In conclusion, the various actions carried out yielded an improvement in pain management, especially in its assessment. PMID- 19543097 TI - Fracture characteristics predict patient mortality after blunt force cervical trauma. AB - Fractures of the cervical spine after blunt cervical trauma are associated with high rates of patient mortality. The purpose of this study was to investigate patient and fracture characteristics that predict mortality. This is a retrospective, case cohort study of all adult patients admitted to our institution between January 1998 and June 2008 with cervical fracture after blunt cervical trauma (N=218). All patient records were reviewed. The patient and fracture characteristics and outcome data were stored. Age (P=0.002), involvement of the fourth cervical vertebra (P=0.002), lamina fracture (P=0.001), and a facet fracture (P=0.006) were identified as independent significant risk factors for mortality. In conclusion, mortality is highly affected by patient age, but fracture location and fracture pattern are also predictive of poor patient outcome. Fracture patterns may increase the risk of spinal cord injury at a level that can affect but not eliminate neural control of the diaphragm, which have the worst prognosis for patient mortality. PMID- 19543098 TI - Takotsubo cardiomyopathy associated with diving. AB - Takotsubo cardiomyopathy consists of cardiomyopathy with transient apical ballooning and left ventricular systolic dysfunction in the absence of atheromatous disease of the coronary arteries, accompanied by ECG changes together with elevated cardiac enzymes appearing in a context of emotional or physical stress. A 51-year-old woman was referred to our emergency department for treatment after chest pain associated with acute dyspnoea during diving. On questioning, the patient confirmed that she had twice dived to 35 m without any missed decompression stops and informed us that she had experienced tightening of the chest followed by sudden dyspnoea during her second ascent in a setting of marked emotional stress since the previous evening. Her ECG showed inverted T waves in DI, aVL, V1 and V2, whereas chest radiographs revealed bilateral infiltration of the lower half of the lung fields and a globular heart. Laboratory tests revealed: troponin Ic 7.49 MUg/l, myoglobin 206 MUg/l, creatine phosphokinase 341 IU/l and N-terminal prohormone brain natriuretic peptide 7919 ng/l. Echocardiography performed in the emergency department showed left ventricular hypokinesia in the medioapical segment with an appearance of apical ballooning, hyperkinesia of the basal portion and a left ventricular ejection fraction of 35%. Coronary angiography revealed healthy coronary arteries. In conclusion, diagnosis of takotsubo cardiomyopathy is based on the guidelines issued at a consensus conference of the Idiopathic Cardiomyopathy Research Committee. This case shows the possibility of this syndrome occurring while diving. PMID- 19543099 TI - The Page kidney phenomenon secondary to a traumatic fall. AB - Page kidney is a rare phenomenon of hyperreninemic hypertension caused by compression of the renal parenchyma. It has been reported in healthy individuals after blunt abdominal or flank trauma, and in patients after invasive nephrological interventions. We present a case of acute on chronic renal failure and Page kidney phenomenon in an elderly male after a traumatic fall, who underwent effective medical management until spontaneous recovery to baseline was observed. A brief discussion on the Page kidney phenomenon is provided with a suggested algorithmic approach towards the management of this process. PMID- 19543100 TI - Inflammatory bowel disease in children: the role of a positive family history. AB - OBJECTIVES: The aim of this study was to evaluate any potential influence of a family history of inflammatory bowel disease (IBD) on the clinical phenotypes and the course of IBD in children. METHODS: In this retrospective study, the notes of 411 children with the diagnosis of IBD, 244 (59.4%) with ulcerative colitis, 129 (31.4%) with Crohn's disease and 38 (9.2%) with IBD unclassified, who were admitted to our department between 1 January 1981 and 31 December 2007 were reviewed. The aim was to assess the prevalence of familial IBD and its impact on the age of disease onset, clinical phenotypes according to the Montreal classification, course and outcome of disease. The control group consisted of IBD children without a family history of IBD, who were admitted to the hospital during the same time period. RESULTS: Thirty five (8.5%) children had a family history of IBD, (ulcerative colitis 6.6%, Crohn's disease 10.9% and IBD unclassified 13.2%). Sixty-eight percent of the 22 pairs of first-degree relatives were concordant for the clinical phenotype of disease. Significantly, more children with familial IBD had symptom onset and/or disease diagnosis before 5 years of age compared with sporadic IBD (P = 0.01 and P = 0.014, respectively); however, no differences were seen in sex, clinical phenotypes, need for aggressive treatment and/or surgery. CONCLUSION: Children with familial IBD had earlier onset of disease compared with those with sporadic IBD. However, this had no significant impact on the clinical phenotypes, the course and/or the outcome of disease. PMID- 19543101 TI - Development of a Web-based concept for patients with ulcerative colitis and 5 aminosalicylic acid treatment. AB - BACKGROUND: Ulcerative colitis (UC) is a lifelong disease with increasing incidence. UC requires frequent outpatient clinic visits and continuous medical treatment. Web-based self-management in other chronic diseases influences disease course, and increases self-adherence, compliance and quality of life (QoL). Lack of easy access to inflammatory bowel disease clinics and patient education, their understanding of the importance of early treatment at relapse, poor compliance and self-adherence can be partly solved by a newly developed Web-based concept. AIMS: To describe the development and validation of the Web-based 'Constant-Care' concept. METHODS: A Web-based treatment program (www.constant-care.dk) and a Patient Educational Centre for UC patients were developed. The feasibility and acceptance of the concept was validated before (group A) and 6 months after (group B) the start of a randomized controlled trial. Patients' level of disease specific knowledge, QoL, anxiety and depression were evaluated. RESULTS: Ten (group A) and 11 (group B) patients fulfilling the diagnostic criteria of mild-to moderate UC participated in the study. All patients reported an ability to initiate self-treatment after the educational training (ET). A significant increase in knowledge from 36 to 69% (group A) and 28 to 75% (group B) was obtained. A majority of the patients were satisfied with the ET. Patients' QoL, anxiety, depression and general well-being showed no difference after the ET. CONCLUSION: Patient education and training through a Web-based program (www.constant-care.dk) seems to be a feasible concept for increasing patients' ability to self-initiate treatment and increase the level of disease-specific knowledge. Relevant adjustment of the concept was implemented. The final outcome of the 'Constant-Care' concept is pending. PMID- 19543102 TI - Modulation of fecal polyamines by viewing humorous films in patients with atopic dermatitis. AB - BACKGROUND/OBJECTIVE: Alteration of intestinal flora was involved in the pathogenesis of atopic dermatitis. Patients with atopic dermatitis were less colonized with Lactobacilli or Bifidobacterium, whereas they were more colonized with Staphylococcus aureus or Enterobacteria. Consequently, fecal levels of bacterial metabolite (polyamines) were reduced. In contrast, stress also induced intestinal mucosal dysfunction against bacteria and impaired intestinal barrier function. We studied the effect of relaxation by viewing humorous films on fecal flora and fecal levels of polyamines. METHODS: Twenty-four healthy individuals and 24 patients with atopic dermatitis either viewed seven control nonhumorous films or seven humorous films sequentially for 7 days. Before and after viewing, feces were obtained, and fecal flora and fecal levels of polyamines were assessed. RESULTS: Neither viewing humorous films nor viewing control nonhumorous films had any effect on healthy individuals. In contrast, viewing humorous films (i) increased colonization with lactobacilli and bifidobacterium, (ii) decreased colonization with S. aureus and Enterobacteria, and (iii) increased fecal levels of polyamines; whereas viewing control nonhumorous films failed to do so in patients with atopic dermatitis. CONCLUSION: Viewing humorous films may modulate fecal levels of polyamines by restoring intestinal flora in atopic dermatitis. PMID- 19543103 TI - Evaluation of gastric fundic and oesophageal varices by 64-row multidetector computed tomography before and after transjugular intrahepatic portosystemic shunt with concurrent left gastric vein embolization. AB - OBJECTIVE: To determine the utility of 64-row multidetector computed tomography (MDCT) in assessing the changes of gastric fundic and oesophageal varices before and after transjugular intrahepatic portosystemic shunt (TIPS) with concurrent left gastric vein embolization. METHODS: Twenty-eight consecutive patients with gastric fundic and oesophageal varices resulted from posthepatitic cirrhosis, clinically confirmed with endoscopy, were enrolled into our study. All the patients underwent thoracicoabdominal triphasic enhancement scans with 64-row MDCT 2 weeks before TIPS along with embolizing left gastric vein, and 6 months after the therapy. Before and after the therapy, the varices shown on multiplanar reconstruction (MPR) were compared by Fisher's exact test to evaluate the changes. RESULTS: Gastric fundic and oesophageal varices were shown in 26 patients (92.9%) and 25 (89.3%) on MPR before the therapy, respectively. After the therapy, these varices diminished markedly, and were illustrated in six (21.4%) and 10 patients (35.7%), respectively (all P<0.05). As for the main inflowing vessel of the varices (left gastric vein), it was illustrated in 28 patients (100.0%) on preoperative CT-MPR. After the embolization, the left gastric vein had been completely occluded on postoperative CT-MPR. CONCLUSION: Significant eradication of gastric fundic and oesophageal varices on follow-up CT MPR portography can be considered to assess the changes of gastric fundic and oesophageal varices before and after TIPS with concurrent left gastric vein embolization, and 64-row MDCT can be an effective modality to evaluate the changes. PMID- 19543104 TI - Chronic Budd-Chiari syndrome as a rare complication of Crohn's disease: a case report. AB - The Budd-Chiari syndrome (BCS) is characterized by hepatic venous outflow obstruction involving the hepatic veins, inferior vena cava, or both. BCS has occasionally been reported in the literature as a very rare complication of ulcerative colitis. However, association of Crohn's disease (CD) and BCS is extremely rare with only a single case reported in the world literature to date. We report a case of a young woman with chronically active, therapy-resistant CD who developed massive ascites, elevation of liver enzymes, and coagulopathy in the course of her disease. She was subsequently diagnosed with BCS for which a successful liver transplantation was performed. Chronically active therapy resistant CD and methylenetetrahydrofolate reductase gene mutation have been identified as possible risk factors for development of BCS in this patient. PMID- 19543105 TI - Internet use among inflammatory bowel disease patients: an Italian multicenter survey. AB - BACKGROUND: The internet has been increasingly used as a resource for accessing health-related information. A recent US survey found that approximately half of inflammatory bowel disease (IBD) patients in an IBD clinic use the internet to gather IBD-specific information. AIM: The aim of this study was to evaluate the use of the internet among Italian IBD patients. METHODS: The study was performed in seven Italian IBD referral centers by using a 28-item anonymous questionnaire. RESULTS: In all, 495 questionnaires were returned for analysis, in which 305 of 495 patients (61.6%) indicated that they are able to access the internet. A large proportion used the internet to gather health-related information (180 of 305, 59.1%) and IBD-related information (226 of 305, 74.2%). The use of the internet increased significantly with level of education (P<0.0001) and household income (P<0.0001). In addition, the use of the internet to gather IBD-related information increased significantly with the increase of disease activity and severity. CONCLUSION: Approximately half of the patients in Italian IBD referral centers used the internet to gather IBD-related information. This use positively correlated with disease activity and severity. The great majority of patients indicated that it was very important for IBD referral centers to have their own IBD-dedicated website. PMID- 19543106 TI - Left ventricular trabeculae: quantification in different cardiac diseases and impact on left ventricular morphological and functional parameters assessed with cardiac magnetic resonance. AB - OBJECTIVES: Left ventricle trabeculae (LVT) are frequently seen in different cardiac diseases. Normal reference values of LVT in different cardiac conditions are not known. The aim of the study was to quantify with cardiac magnetic resonance (CMR), LVT mass (LVTM) and LVTM percentage (LVTM%) in different heart diseases and to evaluate their influence on left ventricular morphological and functional parameters. METHODS: Fifty-nine patients (14 controls, 17 ischemic cardiomyopathy, 15 nonischemic dilated cardiomyopathy, 7 valvular heart disease and 6 with left ventricle hypertrophy) were enrolled. Cine-MR images were acquired with steady-state free-precession sequence in a short-axis view. LVTM was calculated as the difference between LVM excluding/including trabecuale from the blood cavity. LVTM% was calculated as the percentage of the whole left ventricle mass excluding trabeculae from the blood cavity. RESULTS: Mean age was 47.60 +/- 22.03 years; male 62.7%. Mean LVTM was of 33.38 +/- 16.1 g with mean LVTM% of 19.22 +/- 6.5%. Significant differences between groups for both parameters with P values of 0.02 were obtained. Nonischemic dilated cardiomyopathy showed the highest degree of LVTM (44.73 +/- 16.0 g) and LVTM% (23.26 +/- 6%). Significant differences were noted in left ventricular morphological and functional parameters with inclusion/exclusion of LVT in the myocardial mass. CONCLUSIONS: Reference values and differences of LVTM and LVTM% in various cardiac conditions are given for the first time. Quantification of these parameters with CMR may be clinically useful in the differential diagnosis between left ventricular noncompaction and other cardiac diseases. Exclusion of LVT from myocardium alters left ventricular morphological and functional parameters, which have significant clinical importance. PMID- 19543107 TI - Persistent atrial fibrillation associated with syncope due to orthostatic hypotension: a case report. AB - A 40-year-old man was referred to our Cardiology Unit because of three syncopal episodes. The ECG showed atrial fibrillation with a mean heart rate of 104 bpm. The patient was unable to maintain the upright position because of marked orthostatic hypotension associated with bradycardia and presyncope. Two days later the patient underwent direct current shock with sinus rhythm restoration. After the procedure both symptoms and orthostatic hypotension disappeared. A tilt test induced marked hypotension and bradycardia only after nitroglycerine. This is the first report of a patient with persistent atrial fibrillation associated with syncope caused by orthostatic hypotension and bradycardia. The patient's clinical features and positive tilt test only after nitroglycerine lead us to hypothesize that the arrhythmia plays a more important role in the etiology of symptoms with respect to autonomic impairment and that the pathophysiological mechanism responsible for these symptoms could be mainly the baroreceptor reflex. PMID- 19543108 TI - Endovascular repair for isolated iliac artery aneurysms: case report and review of the current literature. AB - For a long time, open surgical repair has been considered the preferred management option for the exclusion of isolated iliac artery aneurysms.The development of transluminally placed endovascular stent grafts, which have been studied extensively for aneurysm exclusion in the abdominal and thoracic aorta, provided a less invasive approach for exclusion of iliac artery aneurysms as compared with surgical reconstruction or open surgical graft placement.Here we report a case of concomitant bilateral common and internal iliac artery aneurysms excluded successfully with multiple stent-grafting deployment with extension to the external iliac arteries and without coil embolization requirements. PMID- 19543109 TI - Mesenteric vasculitis as the initial presentation in children with systemic lupus erythematosus. PMID- 19543110 TI - Introduction of complementary feeding in 5 European countries. AB - OBJECTIVES: Little is known about the practice of introducing complementary feeding across Europe. We aim at describing times of solid introduction in healthy infants in 5 European countries. MATERIALS AND METHODS: Between October 2002 and June 2004, 1678 healthy term infants were either breast-fed (BF) for at least 4 months (n = 588) or study formula-fed (FF) (n = 1090) with different protein contents. Three-day-weighed food protocols were obtained at ages 1, 2, 3, 4, 5, 6, 7, 8, 9, and 12 completed months. RESULTS: Solids were introduced earlier in FF infants (median 19 weeks, interquartile range 17-21) than BF infants (median 21 weeks, interquartile range 19-24). Some 37.2% of FF infants and 17.2% of BF infants received solid foods at 4 completed months, which is earlier than recommended in Europe. Solids had been introduced at 7 completed months in 99.3% of FF infants and 97.7% of BF infants, respectively. Belgium had the highest percentage of solids feeding in FF infants at 3 (15.8%) and 4 (55.6%) completed months, and in BF infants at 4 (43%) and 5 (84.8%) completed months. Multiple regression showed low maternal age, low education level, and maternal smoking to predictors an early introduction of solids at 3 and 4 completed months. CONCLUSIONS: Complementary feeding is introduced earlier than recommended in a sizeable number of infants, particularly among FF infants. Country- and population-specific approaches to adequately inform parents should be explored. PMID- 19543111 TI - Moving from tube to oral feeding in medically fragile nonverbal toddlers. AB - The current article describes a 14-week outpatient protocol for transitioning from gastrostomy tube to oral feeding in toddlers with medical complications. The team ensured that eating skills were mastered before treating patients for 8 weeks with continuous gastrojejunal drip tube feedings and low-dose tricyclic antidepressant and/or gabapentin. We prescribed 6 weeks of megestrol for hunger provocation while withdrawing tube feedings. A chart review after treatment demonstrated 9 subjects were eating exclusively orally and 1 was eating 50% orally. PMID- 19543112 TI - Do some adolescents with rumination syndrome have "supragastric vomiting"? PMID- 19543113 TI - Association between genotypes and phenotypes in coeliac disease. AB - BACKGROUND: Coeliac disease (CD) is a genetically driven immunological intolerance to dietary gluten with a wide range of clinical presentations. The aim of this study was to investigate the heritability of the phenotype in CD and the influence on the phenotype of different genes associated with the disease. PATIENTS AND METHODS: One hundred and seven families with at least 2 siblings with CD were collected. The patients were grouped in symptom grades on the basis of the clinical presentation, the age at diagnosis, and sex. Stratification analyses of the human leucocyte antigen-DQA1 and human leucocyte antigen-DQB1 genotypes, the CTLA4 +49A/G polymorphism, the CTLA4 haplotype MH30*G: 1147*T:+49*A:CT60*G:CT61*A, and the 5q31-33 loci were done. RESULTS: The heritability of the phenotype was estimated to be 0.45. Significant association and linkage was found between the clinical presentation and the CTLA4 +49A/G polymorphism but not for the other genotypes. No correlation was found between genotypes and age at diagnosis or sex. CONCLUSIONS: Our results indicate that the heritability is determiner of the phenotype in CD. The CTLA4 +49A/G polymorphism is correlated to the clinical presentation: the AA genotype is associated with clinically silent disease. PMID- 19543115 TI - Headache in pediatric patients with celiac disease and its prevalence as a diagnostic clue. AB - OBJECTIVES: To establish the prevalence of headache in children with celiac disease (CD), the response to a gluten-free diet, and the prevalence of CD in children affected by headache. METHODS: This hospital-based study included 2 steps. In the retrospective part, 354 children with CD answered a questionnaire investigating the presence of headache before and after the gluten-free diet. The same questionnaire was administered to 200 healthy children matched for sex and age. In the prospective part, 79 children affected by headache were screened for CD by antitransglutaminase IgA. Diagnosis of CD was confirmed by duodenal biopsy; before starting a gluten-free diet patients underwent a brain positron emission tomography study. After 6 months of follow-up children were reevaluated for the presence of headache. RESULTS: Overall, 88 patients with CD complained of headaches before the diagnosis of CD as compared with 16 in the control group (24.8% vs 8%, P < 0.001). After the institution of a gluten-free diet, the headaches significantly improved in 68 children (77.3%), of whom 24 (27.3%) were headache-free during the study period. Four of 79 (5%) headache patients were found to have CD compared with 0.6% of the general population (P = 0.005). The brain positron emission tomography studies did not show any anomalies. During the follow-up, headaches improved in all 4 children with CD. CONCLUSIONS: We recorded -- in our geographical area -- a high frequency of headaches in patients with CD and vice versa with a beneficial effect of a gluten-free diet. Screening for CD could be advised in the diagnostic work-up of patients with headache. PMID- 19543116 TI - Growth assessment of paediatric patients with CF comparing different auxologic indicators: A multicentre Italian study. AB - OBJECTIVES: To evaluate growth in Italian patients with cystic fibrosis (CF). PATIENTS AND METHODS: A multicentre cross-sectional study was carried out on patients with CF attending Italian reference centres. Anthropometric data were evaluated using the Centers for Disease Control and Prevention 2000 reference data. Nutritional failure was defined as height-for-age percentile (HAP) <5th (all patients); weight-for-length percentile (WLP) <10th (patients <2 years); body mass index percentile (BMIp) <15th (patients between 2 and 18 years). The risk of malnutrition (defined as HAP, WLP, and BMIp <25th) and the proportion of patients below the "BMIp goal" (BMIp > or =50th) were also evaluated. Nutritional status was evaluated in the whole population and in relation to age, sex, pancreatic insufficiency, meconium ileus, and lung function. RESULTS: A total of 892 patients with CF (50.7% males, mean age 9.2 years, range 0.1-18 years) were enrolled. The proportion of children with HAP <5th, WLP<10th and BMIp<15th was 12.2%. 12.9%, 20.9%, respectively, and 54.4% did not fulfill the BMIp > or =50th goal. HAP <25th identified the highest proportion of children at risk of malnutrition, whereas BMIp <15th identified the highest proportion of children with nutritional failure. Whatever the criterion used to define malnutrition, the highest proportion of children with nutritional failure was found in adolescence (11-18 years). z scores for height, weight, and BMI were significantly associated with pancreatic status and lung function. Differences among centres for the auxologic parameters were not significant, except for BMIp. CONCLUSIONS: Nutritional failure is present in a minority of Italian patients with CF, particularly during adolescence. Different auxologic indicators should be used for identifying children at risk for or with actual malnutrition. PMID- 19543118 TI - A rare variant of tibial hemimelia and its treatment. AB - Tibial hemimelia is a rare condition. We present a variant of tibial hemimelia not reported in literature before. In this variant, we were able to successfully salvage the limb with excellent function to carry on with life. PMID- 19543117 TI - Feasibility of video capsule endoscopy in the management of children with Peutz Jeghers syndrome: a blinded comparison with barium enterography for the detection of small bowel polyps. AB - OBJECTIVES: Peutz-Jeghers syndrome (PJS) in children may present with anaemia, intussusception, or obstruction from an early age and surgery is common. Prophylactic polypectomy may reduce subsequent complications. Traditional barium enterography (BE) has poor sensitivity and requires significant radiation. We compared the performance of capsule endoscopy (CE) with BE in children with PJS. MATERIALS AND METHODS: Children with PJS (ages 6.0-16.5 years) were prospectively recruited and underwent BE followed by CE, each reported by expert reviewers blinded to the alternate modality. Number of "significant" (>10 mm) and total number of polyps were recorded. Child preference was assessed using a visual analogue questionnaire. Definitive findings were assessed at laparotomy or enteroscopy, when performed. RESULTS: There was no significant difference for >10 mm polyp detection. Six polyps were found in 3 children by both modalities: 3 polyps in 2 children at CE, 3 polyps in 1 child at BE (P=0.50). Re-review of 1 CE identified 3 polyps that were missed in 1 child at initial reading. Significantly more <10 mm polyps were identified by CE than BE: 61 vs 6 (P=0.02). CE was significantly more comfortable than BE (median score CE 76 [interquartile range 69-87] vs BE 37 [interquartile range 31-68], P=0.03) and was the preferred investigation in 90% (P=0.02). CONCLUSIONS: CE is a feasible, safe, and sensitive test for small bowel polyp surveillance in children with PJS. It is significantly more comfortable than BE and is the preferred test of most children for future surveillance. There is a learning curve for reporting CE studies in PJS and appropriate training is essential. PMID- 19543114 TI - Randomized controlled trial of feeding a concentrated formula to infants born to women infected by human immunodeficiency virus. AB - OBJECTIVE: We tested the hypothesis that concentrated formula (CF) begun within the first 2 weeks of life increases growth in infants born to human immunodeficiency virus (HIV)-infected mothers. MATERIALS AND METHODS: HIV-exposed infants from the United States, the Bahamas, and Brazil were randomized in a double-blind, controlled trial to receive either a CF (87 kcal/100 mL [26 kcal/oz]) or a standard formula (SF; 67 kcal/100 mL [20 kcal/oz]) for 8 weeks. This article presents results for infants who were not determined to be HIV infected based on testing at 4 weeks. Primary outcomes were safety, tolerability, and growth in weight and length. RESULTS: Two thousand ninety-seven infants were enrolled, of whom 1998 were uninfected and had study formula dispensed. At weeks 4 and 8, uninfected infants receiving CF showed higher energy intake than those who were receiving SF (P < 0.001). By week 8, uninfected infants assigned to CF weighed more than infants receiving SF. There were no consistent differences in measures of tolerability, and rates of discontinuation or perceived formula intolerance were similar between treatment groups. CONCLUSIONS: A CF is well tolerated and results in increased weight gain compared with SF. Until the HIV status of an infant is reliably determined, early introduction of a CF in HIV exposed children may have beneficial effects on growth. The role of early nutritional intervention remains to be determined for individuals living in countries with endemic malnutrition for whom formula feeding is a viable option. PMID- 19543119 TI - Bronchioloalveolar carcinoma (lepidic growth) component is a more useful prognostic factor than lymph node metastasis. AB - INTRODUCTION: Although many factors predictive of patient survival have been reported for lung cancer, no comparative studies have attempted to determine those that are most significant for practical medicine. METHODS: We conducted a retrospective review of 139 patients who underwent complete resection of adenocarcinomas less than 2 cm in diameter between 1993 and 2000 at the National Cancer Center Hospital (Tokyo, Japan). The MIB-1 labeling index (LI), immunohistochemical staining for carcinoembryonic antigen (CEA), p53, p27, epidermal growth factor receptor (EGFR), phosphorylated-EGFR (pEGFR), Cox-2, neuronatin, gammaH2AX, and thyroid transcription factor-1 (TTF-1), the prevalence of a micropapillary pattern, and the ratio of the bronchioloalveolar cell carcinoma (BAC) or lepidic growth (LG) component were determined, and their significance as prognostic factors for lung adenocarcinoma was compared. RESULTS: Univariate analysis showed that lymph node metastasis (p-N status), BAC/LG component, vascular invasion (p-V status), MIB-1 LI, pEGFR, and CEA were prognostically significant (p-N status: p < 0.0001, BAC/LG: p = 0.0005, p-V status: 0.002, MIB-1 LI: p = 0.005, pEGFR: p = 0.024, and CEA: p = 0.049). Multivariate analysis showed that only p-N status (p = 0.013) was of prognostic significance. However, BAC/LG component (p = 0.051) was a more reliable prognostic factor than p-N status in mixed adenocarcinoma with a BAC/LG component. CONCLUSION: In comparison with other immunohistochemical and histopathologic factors, BAC/LG component is independently and reliably prognostic for small adenocarcinoma of the lung, and, in particular, for the major histologic subtype (adenocarcinoma mixed subtype with BAC/LG), BAC/LG component is more reliably prognostic than lymph node metastasis. PMID- 19543120 TI - Why consider patients' preferences? A discourse analysis of clinical practice guideline developers. AB - BACKGROUND: Several organizations are advocating for patients' preferences to be considered in clinical practice guideline development and implementation. However, lack of agreement on the goal and meaning of this policy curtails evaluation and development of patient involvement programs. GOAL: To describe guideline developers' discourses on the goal of considering patients' preferences. METHOD: DESIGN: Qualitative study using discourse analysis. SUBJECTS: 18 participants (patients, health professionals, and public health experts) from 2 groups of British guideline developers. DATA COLLECTION AND ANALYSIS: Template analysis of semi-structured individual interviews was strengthened by active search for deviant cases, team debriefing, and member checking. RESULTS: All respondents supported the idea of taking account of patients' preferences in guidelines. Divergences with the goal and meaning of considering preferences were structured in 4 discourses: (1) The Governance discourse constructs guideline development as a rational process of synthesizing population data-including evidence on patients' preferences-to maximize public health within the constraints of available resources; (2) the Informed Decision discourse aims at fostering patients' choice by providing tailored information on the risks and benefits of interventions; (3) the Professional Care discourse insists on basing professionals' recommendations on the individual characteristics of patients; (4) The Consumer Advocacy discourse argues for greater political power and influence over guideline development and clinical decision making. CONCLUSIONS: The identified discourses provide a set of hypothesis on how patient involvement programs are expected to work, which could help clarify the goals pursued by guideline organizations and anchor further evaluation efforts. PMID- 19543122 TI - The relationship between perceived practice quality and quality improvement activities and physician practice dissatisfaction, professional isolation, and work-life stress. AB - BACKGROUND: The importance of physician well-being has been well-documented. However, little is known about how physicians' self-reported quality improvement (QI) activities and quality of care are related to their practice dissatisfaction, professional isolation, and work-life stress. METHODS: We surveyed a random sample of 1884 physicians in Massachusetts by mail and assessed their practices' participation in QI activities and quality of care, as well as their feelings of professional isolation, work-life stress, and practice dissatisfaction. RESULTS: A total of 1345 physicians responded (71.4% response rate). Most respondents reported QI activities in their practices (85%) and subsequent evaluation of these activities (62%). Approximately one-third (33%) reported quality problems in their practice. In linear regression analyses, the presence of quality problems was independently associated with increased professional isolation, work-life stress, and practice dissatisfaction. In contrast, physicians from practices that were involved in the evaluation of QI activities had significantly less isolation, stress, and dissatisfaction. Participation in QI activities was also independently associated with less dissatisfaction. A substantial fraction of physicians reported moderate to severe problems with isolation (17%), work-life stress (31%), and dissatisfaction (27%). CONCLUSIONS: Substantial practice dissatisfaction, professional isolation, and work-life stress are experienced by physicians and they seem to be inversely correlated with QI activities. Physicians who perceive quality problems in their practices are more likely to experience dissatisfaction, isolation, and stress. Efforts to engage physicians in QI and systems change should assess how these programs affect physicians themselves and the care that they deliver. PMID- 19543121 TI - Differences between individual and societal health state valuations: any link with personality? AB - OBJECTIVE: The concept of "adaptation" has been proposed to account for differences between individual and societal valuations of specific health states in patients with chronic diseases. Little is known about psychological indices of adaptational capacity, which may predict differences in individual and societal valuations of health states. We investigated whether such differences were partially explained by personality traits in chronic disease patients. RESEARCH DESIGN: Analysis of baseline data of randomized controlled trial. SUBJECTS: Three hundred seventy patients with chronic disease. MEASURES: The NEO-five factor inventory measure of personality, EuroQoL-5D (EQ-5D) societal-based, and the EQ visual analogue scale individually-based measures of health valuation. RESULTS: Regression analyses modeled Dev, a measure of difference between the EQ-Visual Analogue Scale and EQ-5D, as a function of personality traits, sociodemographic factors, and chronic diseases. Individual valuations were significantly and clinically higher than societal valuations among patients in the second and third quartile of conscientiousness (Dev = 0.08, P = 0.01); among covariates, only depression (Dev = -0.04, P = 0.046) was also associated with Dev. CONCLUSION: Compared with societal valuations of a given health state, persons at higher quartiles of conscientiousness report less disutility associated with poor health. The effect is roughly twice that of some estimates of minimally important clinical differences on the EQ-5D and of depression. Although useful at the aggregate level, societal preference measures may systematically undervalue the health states of more conscientious individuals. Future work should examine the impact this has on individual patient outcome evaluation in clinical studies. PMID- 19543123 TI - Quality assessments by sick and healthy beneficiaries in traditional Medicare and Medicare managed care. AB - BACKGROUND: The Centers for Medicare & Medicaid Services pays for services provided through traditional fee-for-service (FFS) Medicare and managed care plans (Medicare Advantage [MA]). It is important to understand how financing and organizational arrangements relate to quality of care. OBJECTIVES: To compare care experiences and preventive services receipt in traditional Medicare and MA for healthy and sick beneficiaries. METHODS: Randomly selected beneficiaries responded to the 2003 and 2004 Consumer Assessments of Healthcare Providers and Systems (CAHPS(R)) surveys. We analyzed 237,221 MA responses (80% response rate) and 153,535 from FFS (68% response rate). We compared case-mix-adjusted CAHPS scores between FFS and MA for healthy and sick beneficiaries on 7 CAHPS measures of care experiences and 3 preventive service measures. RESULTS: CAHPS scores were lower in MA than FFS for all care experience measures except office wait time. The sick had less favorable care experiences than the healthy for all measures, but were more likely to receive each preventive service (P < 0.001). FFS-MA differences were larger for the sick than the healthy for 5 of 7 experience measures (P < 0.05), and were twice as large for physician ratings and interactions. Office wait time and rates of immunization were better in MA than FFS (P < 0.001), with no differences between healthy and sick groups. CONCLUSIONS: Beneficiaries in health plans report less favorable care experiences than those in FFS, particularly among the sick, but preventive service measures are higher in MA. The Centers for Medicare and Medicaid Services should strengthen efforts to improve care experiences of the sick, particularly in MA, and preventive service receipt in FFS. PMID- 19543124 TI - Patient, physician, and consumer drivers: referrals for short stature and access to specialty drugs. AB - BACKGROUND: Candidates for specialty drugs, the fastest growing and costliest pharmaceuticals, typically originate with primary care referrals. However, little is known about what drives such referrals-especially for large populations such as short, otherwise normal children (idiopathic short stature). Recent expanded approval of growth hormone (GH) makes more than 585,000 US children eligible for such treatment, potentially costing over $11 billion/y. METHODS: To quantify the relative impact of patient physiological indicators, physician characteristics, and consumer preferences on referrals to endocrinologists (and potential access to GH) for short children, a national study of 1268 randomly selected US pediatricians was conducted, based on a full factorial experimental design in a structured survey. RESULTS: While patient indicators (height, growth pattern) influenced referrals (P < 0.001), consumer drivers (family concern) and physician attitudes had almost as great an impact-especially for children with less severe growth impairment (P < 0.001). Physician belief that short stature impairs emotional well-being and physician characteristics (female, older, shorter, beliefs about drug company information) increased referrals (P < 0.03-0.001) independent of growth parameters. CONCLUSIONS: Referral recommendations that create the pool of candidates for the specialty drug GH are heavily swayed by physician characteristics and consumer preferences, particularly in the absence of compelling physiological evidence. This makes most of children with short stature strikingly susceptible to nonphysiological influences on referrals that render them candidates for this specialty drug. Only 1 additional referral per US pediatrician would likely increase GH costs by over $100 million/y. PMID- 19543125 TI - Cyclooxygenase-2 immunohistochemistry in human melanoma: differences between results obtained with different antibodies. AB - Several groups have reported that cyclooxygenase-2 (COX-2) expression is significantly enhanced in human melanomas, and that the expression of this protein may be useful as diagnostic and prognostic marker for the disease. At the same time, collective analysis of immunohistochemical data on the COX-2 expression in melanomas, presented by different researchers, shows a clear lack of consistency of reported results commonly assigned to differences in protocols used for the staining. This paper describes a study involving the parallel use of three different primary anti-COX-2 antibodies targeting different COX-2 epitopes. A surprising outcome is that although the three antibodies gave very consistent results for the COX-2 expression in keratinocytes, they showed significant differences in immunoreactivity for both melanocytic naevi and melanomas. This phenomenon has not been described before, and has implications for the selection of antibodies for studies on the diagnostic potential of COX-2 for melanoma. PMID- 19543126 TI - The anatomic distribution of melanoma and relationships with childhood nevus distribution in Colorado. AB - The nature of the relationship between nevus development in childhood and later development of melanoma is unclear. Data on melanoma diagnoses by histologic type and anatomic site were obtained for 2351 white, non-Hispanics in Colorado from the Colorado Central Cancer Registry between 2000 and 2004. Nevus size and body site were ascertained during skin exams conducted in the summer of 2007 on 717 white, non-Hispanic children aged 8-9 years. Chi-square goodness-of-fit analysis was used to assess the association between the anatomic site distributions of nevi versus melanoma. Superficial spreading melanoma was the most frequent histology, followed by lentigo maligna melanoma. Nodular melanoma was the least common histology. For males, there was no significant difference between the distribution of medium-sized (> or =2 mm) nevi and the distribution of both superficial spreading and nodular melanomas. For females, there was no significant difference between the anatomic distribution of small-sized (<2 mm) nevi and the distribution of nodular melanoma, and there was marginal evidence for a difference between the distribution of medium-sized (> or =2 mm) nevi and the distribution of nodular melanoma. There was evidence for a difference between all of the nevus distributions and the distributions of superficial spreading and lentigo maligna melanoma in females. The similarities between the nevus and melanoma distributions are interesting findings, but it is difficult to interpret the significance of these findings based on the current state of knowledge of melanoma etiology. PMID- 19543127 TI - Multiple melanoma arising on a burn scar and extensive sunburn: a case report and a review of the literature. AB - Melanoma arising in a burn scar is very uncommon. We report a recent case of a female patient in whom two different melanomas arose on a wide back burn scar at different times, focusing attention on three different potential risk factors for melanoma the patient had: sunburns, laser therapy on back and burn scar. PMID- 19543128 TI - Life satisfaction, distress, and resiliency across the life span of women. AB - OBJECTIVE: This study aimed to determine (1) the relationship between life satisfaction, mental disorders, and aging in the female community and (2) to identify the impact of vulnerability factors, personal (resilience, self-esteem), and social resources on life satisfaction and distress. METHODS: A stratified random sample of the German female population (N = 2,540) was investigated using standardized questionnaires of life satisfaction (Questions on Life Satisfaction), depression, anxiety (Patient Health Questionnaire), resilience scale (RS-11), and self-esteem (Rosenberg Self-Esteem Scale). RESULTS: When participants were divided into six age groups (18-30, 31-40, 41-50, 51-60, 61-70, and >70 y), we found lower life satisfaction, satisfaction with partnership, and sexuality beyond the age of 50 years. Satisfaction with health was decreased already at early adulthood. Although the importance of partnership/sexuality was reduced around midlife, the importance of health increased. Depression was highest in the oldest group, whereas anxiety remained fairly stable across the cohorts. Life satisfaction was strongly associated with resilience, a good household income, the presence of a partnership, absence of anxiety and depression, lack of unemployment, positive self-esteem, religious affiliation, and-least-age. CONCLUSIONS: Personal and social resources and the absence of anxiety and depression are of crucial importance for the maintenance of life satisfaction in aging women. PMID- 19543129 TI - Bazedoxifene, a selective estrogen receptor modulator: effects on the endometrium, ovaries, and breast from a randomized controlled trial in osteoporotic postmenopausal women. AB - OBJECTIVE: The aim of this study was to evaluate the endometrial, ovarian, and breast safety of bazedoxifene used as a treatment for postmenopausal osteoporosis. METHODS: Healthy women (aged 55-85 y) with osteoporosis were enrolled in a randomized, double-blind, placebo-controlled phase 3 trial. Participants were randomized to treatment with bazedoxifene 20 or 40 mg, raloxifene 60 mg, or placebo daily for 3 years. Endometrial and ovarian safety was assessed by periodic transvaginal ultrasonography and endometrial biopsy through 24 months. Gynecologic and breast-related adverse events were recorded throughout the study. RESULTS: Among 753 participants with available transvaginal ultrasonography data, there were no significant between-group differences in overall endometrial thickness or in the percentage of participants with endometrial thickness greater than 5 mm at 12 or 24 months. Changes in the mean endometrial thickness (+/-SE) from baseline were -0.07 +/- 0.11 mm (bazedoxifene 20 mg), 0.10 +/- 0.11 mm (bazedoxifene 40 mg), 0.16 +/- 0.12 mm (raloxifene 60 mg), and -0.08 +/- 0.11 mm (placebo) at 24 months. There was one report of endometrial hyperplasia in each group, and there were zero, two, two, and three reports of endometrial carcinoma with bazedoxifene 20 and 40 mg, raloxifene 60 mg, and placebo, respectively. There were no clinically important changes from baseline in the number or size of ovarian cysts among groups. There was a significantly lower incidence of fibrocystic breast disease (P or =1: 40%) contrasted with that of subjects spontaneously reporting respiratory problems (9%). Among these, 72% reported limitations in daily-life activities but only 14% considered that they were severely affected by their respiratory status. A very low proportion of subjects knew the term COPD (8%) and only 66% of these associated COPD with a respiratory disease. CONCLUSIONS: There is a wide gap between the high number of subjects at risk of COPD in the general population and the frequency of breathlessness in these subjects on one hand, and the poor knowledge of the disease, poor perception of symptoms, and under-diagnosis and under-use of spirometry on the other. Increasing awareness of COPD in the population is needed. PMID- 19543172 TI - [A stairclimbing test in COPD patients assessment]. AB - Patients suffering from Chronic Obstructive Pulmonary Disease (COPD) are usually evaluated for cardio-respiratory capacity on a flat surface even though these patients generally encounter discomfort only when climbing stairs or ascending a slope. The purpose of this study was to describe the cardio-respiratory evaluation of these patients on climbing stairs. A staircase test was carried out and compared to the 6-min walking test. The maximum duration of ascent was 2 min, covering a maximum of four flights of stairs. The conditions of the staircase test and the instructions given to the patients regarding walking were designed to be consistent with the activities of daily life. We recorded the height reached and the distance covered; we also recorded dyspnoea, oxygen saturation (SpO2) and heart rate (HR); we estimated the maximal oxygen consumption (VO2max) and the physiological cost index (PCI) during each test. The statistical analysis of the results was surprising: it showed that there was no significant difference between the 6-min walking test and the stair climbing test on the following criteria: estimated VO2max, SpO2, and variation in heart rate between rest and effort (DeltaHR); it also showed that these two tests correlated well for dyspnoea (p<0.0001). It appears that the PCI of stair climbing is much higher than the PCI of walking on a level surface. These results may be explained partly by the anaerobic metabolic pathway used predominantly by patients with COPD in brief periods of exertion and partly by the muscular dysfunction which also accompanies brief exertion. The use of the PCI adds greater precision to the interpretation of the results of cardio-respiratory evaluation of COPD patients. The value of this index is proportional to the incapacity described by these patients. The staircase test is complementary to the 6-min walking test, making it possible to evaluate restriction of activity in COPD patients. Its implementation is simple and accessible. PMID- 19543173 TI - [Practical methods for controlled diaphragmatic breathing in 2009: proposals for a consensus]. AB - BACKGROUND: This work has been completed at the request of the French Language Society of Pneumology. It is the result of collaboration between the 'Muscles and Respiration Group' and the 'Working Group in Physiotherapy', arriving at a consensus on diaphragmatic breathing. RESULTS: From the literature followed by a formalized consensus methodology, the authors specify the terminology and define the appropriate methods of this technique that should be used. CONCLUSION: Analysis of the literature to date does not allow assessment of the efficacy of this technique. The precise definition of the methods of diaphragmatic breathing as proposed in the present study will be used as a basis for forthcoming studies on this technique. PMID- 19543174 TI - [Remarkable evolution of hepatic, pleural and pulmonary amoebiasis: a case report]. AB - Entamoeba histolytica, is a human pathogen which is endemic in tropical areas. The most common extra-intestinal locations for disease after the liver are pleural and pulmonary. Although the outcome of pleural and pulmonary amoebiasis is generally favourable, it is important to note that a significant proportion of patients do develop pleural and pulmonary after-effects. We report a case of pleural and pulmonary amoebiasis where the evolution of a massive pleural effusion and 3 associated liver abscesses was spectacular. The surgical management of pleural, pulmonary and liver amoebiasis is discussed. PMID- 19543175 TI - [Respiratory consequences of inhalation of adulterated cannabis]. AB - Cannabis is widely smoked in Europe and its increasing use is becoming a major public health problem. Adulterating cannabis with glass beads or sand is a new trick used by dealers to increase the weight and boost profits. These recent practices are not without danger. We report two cases of respiratory symptoms related to the use of this kind of adulterated cannabis. The first case is a 33 year-old patient admitted for an acute inhalation pneumonitis secondary to smoking cannabis adulterated with grit sand. The CT scan showed patchy ground glass opacities, mainly in the upper lobes. A broncho-alveolar lavage, examined under polarized light, revealed birefringent intracellular particles, identified as silica, in alveolar macrophages. Spontaneously clinical and radiological improvements were observed after stopping the use of contaminated cannabis. The second patient, who smoked cannabis mixed with glass beads, described epistaxis, mouth ulcers, sore throats and cough. CT scan and BAL were normal. Adulteration of cannabis with microscopic glass beads, alone or mixed with sand, is a recent and widespread practice in Europe. These anecdotal reports prompted the French Department of Health to advise cannabis smokers of the harmfulness of these contaminants. PMID- 19543176 TI - [Allergic asthma due to sausage mould]. AB - INTRODUCTION: Chronic cough is a common reason for consultation in respiratory medicine. The three most frequent causes of chronic cough are asthma, postnasal drip syndrome or rhinosinusitis, and gastro-oesophageal reflux disease. CASE REPORT: We describe a case of chronic cough related to occupational asthma with sensitivity to dry sausage mould (Penicillium nalgiovensis) in a worker in a semi industrial pork butchers. The diagnosis was based on the history of symptoms, positive skin prick-tests, spirometry, a normal pulmonary CT-scan, and a favourable outcome after avoidance of the allergen. It was notified as a case of occupational disease. CONCLUSION: A diagnosis of asthma should be considered in a worker exposed to dry sausage mould presenting with respiratory symptoms like chronic cough without evidence of hypersensitivity pneumonitis. Skin prick-tests and allergen avoidance are useful in the diagnosis. PMID- 19543177 TI - [An infrequent tumor of the posterior and inferior mediastinum]. PMID- 19543187 TI - Diabetic limb salvage. PMID- 19543188 TI - Diabetic patients: epidemiology and global impact. AB - Definition of the exact epidemiology and the global impact of diabetes is not easy, being strictly related to the availability of data in developing countries and to the use in the existing population-based investigations of common criteria for the diagnosis and definition of diabetes. According to the World Health Organization (WHO) the total number of people with diabetes was 171 million in 2000, and is projected to rise up to 366 million in 2030. The true prevalence of peripheral arterial disease (PAD) in people with diabetes has been difficult to determine, as most patients are asymptomatic, many do not report their symptoms, screening modalities have not been uniformly agreed upon, and pain perception may be blunted by the presence of peripheral neuropathy. Population-based studies, using a validated and reproducible test, have revealed a prevalence of PAD in people with diabetes to be up to 30%. Among people with diabetes, the annual incidence of developing a foot ulcer ranges from 1% to 4.1% and the prevalence ranges from 4% to 10%, which suggests that the lifetime incidence may be as high as 25%. Foot ulcer associated to PAD requires revascularization, although it is generally considered that the outcome in those people is inferior to that in non diabetic patients. In summary, the increasing worldwide diabetes prevalence will inevitably result in increasing proportions of deaths from cardiovascular disease, as well as in increased prevalence and associated consequences of other complications of diabetes. As suggested by WHO, a concerted, global initiative is required to address the diabetes epidemic. PMID- 19543190 TI - Surgical treatment of the neuroischemic foot. AB - As many as 1% of older people in the Western world are at risk of developing a foot ulcer as a complication of diabetes mellitus. The resultant debility and disability constitutes a burden for both individuals and their health services. When peripheral arterial insufficiency complicates neuropathy there is a tenfold risk of ulceration progressing to infection, gangrene and amputation. Patient education and the vigilant implementation of preventive measures offer the best prospects for containment of the problem. Patients faced with ulceration and limb loss require access to a co-ordinated and comprehensive diabetic foot service offering detailed assessment and a full range of social, medical and surgical therapies. PMID- 19543191 TI - Other endovascular methods of treating the diabetic foot. AB - Critical limb ischemia (CLI) is an endstage manifestation of peripheral artery disease (PAD) and typically describes patients with ischemic rest pain (Rutherford Category 4), or patients with ischemic skin lesions, either ulcers or gangrene (Rutherford Category 5-6). CLI due to infrapopliteal lesions is often not a good indication for infrageniculate bypass surgery placement, due to the presence of prohibitive comorbidities, inadequate conduit, and lack of suitable distal targets for revascularization. Therefore, CLI patients due to blockage of below-the-knee arteries are in benefit of the endovascular approach. Infrapopliteal PTA became feasible with the introduction of low-profile peripheral balloon systems and the use of coronary balloons. Stent implantation, however, is generally reserved for cases with a suboptimal outcome after PTA (i.e. >50% residual stenosis, flow-limiting dissection). Because of the fear that early thrombosis and late luminal loss due to intimal hyperplasia formation will potentially lead to insufficient long-term patency rates, efforts to extend the range of endovascular approaches to CLI are presently focused on. Cryoplasty, scoring balloon angioplasty, excimer laser therapy and atherectomy using different types of atherectomy devices have been introduced. The current article provides an overview of the endovascular treatment strategies for infrapopliteal lesions in patients with CLI. PMID- 19543189 TI - Treatment of diabetic foot ulcers. AB - Diabetic foot ulcers are a major health care problem. Complications of foot ulcers are a leading cause of hospitalization and amputation in diabetic patients. Diabetic ulcers result from neuropathy or ischemia. Neuropathy is characterized by loss of protective sensation and biomechanical abnormalities. Lack of protective sensation allows ulceration in areas of high pressure. Autonomic neuropathy causes dryness of the skin by decreased sweating and therefore vulnerability of the skin to break down. Ischemia is caused by peripheral arterial disease, not by microangiopathy. Poor arterial inflow decreases blood supply to ulcer area and is associated with reduced oxygenation, nutrition and ulcer healing. Necrotic tissue is laden with bacteria apt to grow in such an environment, which also impairs general defence mechanisms against infection. Infections often complicate existing ulcers, but are seldom the cause for ulcers. Protective footwear helps to reduce ulceration in diabetic feet at risk. Relieving pressure on the ulcer area is necessary to allow healing. Blood supply needs to be improved by revascularisation whenever compromised. Systemic antibiotics are helpful in treating acute foot infections, but not uninfected ulcers. Osteomyelitis may underlie a diabetic ulcer and is often treated by resection of the infected bone and always by antibiotics, the mode and length of treatment depending on the adequacy of the debridement. The aim of ulcer bed preparation is to convert the molecular and cellular environment of the chronic ulcer to that of an acute healing wound by debridement, irrigating and cleaning. Moist dressings maintain wound environment favorable for healing. All attempts should be done to prevent diabetic foot ulceration and treat existing ulcers by multidisciplinary teams in order to decrease amputations. Indeed, improvement in ulcer healing has been observed with primary healing rates of 65-85% in mixed series. Even when healed, diabetic foot should be regarded as a life-long condition and treated accordingly to prevent recurrence. Long-term efforts have reduced amputation 37-75% in different European countries over 10-15 years. PMID- 19543192 TI - Below the knee angioplasty among diabetic patients. AB - The treatment of below knee arterial disease has undergone a gradual shift over the last few years to incorporate a greater proportion of endovascular treatments. Not only does this include patients who now have endovascular therapy rather than surgery, but also patients who in the past would have been offered supportive treatment only, due to factors such as being medically unfit for surgery, lacking sufficient donor vein for bypass grafting, or swelling. Diabetes mellitus is becoming increasingly common, potentially causing numerous comorbidities in patients. It tends to have a more distal pattern of peripheral vascular disease, presenting later and with generally high complication and failure rates following therapy (surgical or endovascular) and higher amputation rates. Given these comorbidities, the reduced morbidity and mortality of endovascular treatments may be beneficial in treating below-knee arterial disease. The success and complication rates of endovascular therapy vary between treatment for claudication and critical limb ischaemia, though success rates are improving and in some cases are comparable to the current gold standard of surgical bypass. PMID- 19543193 TI - Clinical results of below-the knee intervention using pedal-plantar loop technique for the revascularization of foot arteries. AB - AIM: Recent registries and randomized trials support the role of percutaneous revascularization in patients with critical limb ischemia (CLI) due to below-the knee (BTK) atherosclerotic disease, as percutaneous transluminal angioplasty (PTA) for BTK disease has shown to be feasible and safe in this setting. Nonetheless, succes rates remain suboptimal with current techniques. The authors aimed to appraise clinical results following PTA of foot vessels exploiting a novel technique, based on the recanalization of both pedal and plantar arteries and their anatomical anastomosis in order to restore direct arterial in-flow from both anterior and posterior tibial vessels, defined as the pedal-plantar loop technique. METHODS: Baseline, procedural and mid-term outcome data of all consecutive patients with CLI due to BTK disease in which PTA was attempted using the pedal-plantar loop technique were prospectively collected between January 2007 and September 2008. The primary end-point was acute success (i.e. the composite of technical, angiographic and procedural success). Secondary end points included limb salvage rate, major (above the ankle) and minor (below the ankle) amputation, change in Rutherford class and transcutaneous oxygen tension, reocclusion/restenosis, rehospitalization, and repeat revascularization after 12 months. RESULTS: A total of 1331 consecutive patients with CLI were treated using BTK PTA and 135 (10.1%) were approached with the pedal-plantar loop technique in order to recanalize the foot arteries. Target lesions were mostly occlusive and diffusely diseased, involving in most cases the tibial arteries as well as the in flow and out-flow vessels. Acute success was achieved for tibial PTA in 100% of the cases, with ability to position and inflate the balloon and achieve adequate angiographic results without peri-procedural complications in all, whereas acute success for the pedal-plantar loop technique was 85%. Clinical improvement in functional status was obtained and maintained after an average of 12 months, with a significant improvement of transcutaneous oxygen tension after 15 days, 59+/-16 mmHg in the group of patients in which the foot arteries revascularization was successfully feasible, versus 42+/-12 mmHg in patients achieving patency of two BTK vessels at the ankle level with partial out-flow in the foot (P<0.001). CONCLUSIONS: Percutaneous revascularization of foot arteries in patients with CLI is feasible and safe, and appears to provide positive clinical results at both acute and mid-term follow-up. PMID- 19543194 TI - Heparin-bonded expanded polytetrafluoroethylene graft for infragenicular bypass: five-year results. AB - AIM: This study assessed five-year results with a heparin-bonded expanded polytetrafluoroethylene (ePTFE) graft (GORE PROPATEN Vascular Graft) used in infragenicular bypasses in patients with peripheral arterial disease (PAD) with no available autologous vein for grafting. METHODS: A retrospective review was conducted of the records of the 75 patients given a heparin-bonded ePTFE graft in 2003 to 2007 and in whom a femoropopliteal 3 (FP 3; distal anastomosis below the knee articulation) or femorocrural (FC) anastomosis was performed. Graft patency rates were determined by life-table analysis. RESULTS: Most patients (88%) had previously been treated for PAD (Rutherford category 4 to 6 in 91% of patients overall); 31% had renal insufficiency; and 55% had only one patent run-off vessel. Three major procedure-related adverse events occurred within 30 days postoperatively: one death from congestive heart failure, one hemorrhage, and one graft infection. The 1-, 2-, 3-, 4-, and 5-year primary patency rates were, respectively, 77.3%, 71.4%, 71.4%, 71.4%, and undeterminable for the FP 3 bypasses and 64.4%, 56.8%, 49.7%, 49.7%, and 49.7% for the FC bypasses. The 1-, 2 , 3-, 4-, and 5-year secondary patency rates were, respectively, 87.7%, 82.5%, 82.5%, 82.5%, and undeterminable for the FP 3 bypasses and 87.1%, 78.0%, 71.5%, 71.5%, and 71.57% for the FC bypasses. The major amputation rate was 16% or limb salvage was 84% at five years. CONCLUSIONS: The heparin-bonded ePTFE graft provided good long-term results in infragenicular bypasses in patients with severe PAD. PMID- 19543195 TI - Carotid highly-calcified de novo stenosis and cutting-balloon angioplasty: a tool to prevent haemodynamic depression? AB - AIM: Severe highly-calcified de-novo lesions of carotid arteries are important predictors of haemodynamic depression (HD) after carotid artery stenting (CAS). Cutting-balloon angioplasty (CBA) using low-inflation pressures has been described as an effective method to achieve important luminal gain and avoid HD. METHODS: from January 2005 to December 2007 102 consecutive patients (mean age 76.36+/-7.54 years) with severe highly-calcified de novo lesions of carotid artery undergoing CAS were prospectively observed and randomized in two groups: group 1 (N=50) underwent standard CAS, group 2 (N=52) underwent CAS with CBA. Patients with prior ipsilateral carotid endarterectomy (CEA), betablockers therapy or arrhytmias were excluded. HD was defined as periprocedural hypotension (systolic blood pressure <90 mmHg) or bradycardia (heart rate <60 beats/min). CBA angioplasty was carried out using CB coronary device, 0.014 inch compatible, from 3 to 4 mm or CB peripheral device, 0.018 inch compatible, from 5 to 6 mm in diameter, inflated at a mean-maximum value of 8.6 atmospheres; the average number of cuts per lesion was 2.7. RESULTS: Demographic and clinical characteristic of both groups at the basal conditions were comparable. HD occurred in 18/50 (36%) procedures in group 1, and in 3/52 (5.76%) in group 2. The difference between the two groups concerning HD incidence was statistically significant (P<0.001). There was a strong (P<0.001) association between HD and CBA and the OR=0.109 (95%CI 0.019-0.425) confirmed the protective role of CBA. No major intraprocedural complications were observed in this series. Postprocedural Doppler ultrasound scan showed one case (2%) of in-stent restenosis in group 1 representing early failure due to recoil, and only 1 (1.92%) neurological adverse event (transient ischemic attack, TIA) in group 2. CONCLUSIONS: HD is a common occurrence after CAS, especially in patients with both long and calcified plaque. Only a tailored procedure with a correct remodelling of the plaque allows to avoid both HD and elastic recoil of the target lesion. PMID- 19543196 TI - Applicability and clinical results of percutaneous transluminal angioplasty with a novel, long, conically shaped balloon dedicated for below-the knee interventions. AB - AIM: The aim of this study was to assess the feasibility, safety and efficacy of percutaneous transluminal angioplasty (PTA) in patients with critical limb ischemia (CLI) using a novel balloon designed for below-the-knee (BTK) indications. METHODS: The authors have prospectively collected baseline, periprocedural and mid-term data of all consecutive patients with CLI due to BTK disease in which PTA was attempted using a long (210 mm), conically-shaped balloon (0.5 mm tapering from proximal to distal balloon edges). The primary objective was the assessment of acute success (composite of technical, angiographic and procedural success). The secondary assessments included limb salvage rate, major (above the ankle) and minor (below the ankle) amputation, change in Rutherford class and cutaneous oxygen tension, reocclusion/restenosis, rehospitalization, and repeat revascularization after one year. RESULTS: A total of 31 patients were treated with 36 long tapered balloons. Ten patients presented with ischemic tissue loss. Target lesions were mostly occlusive and diffuse, commonly involving the tibial arteries as well as the in-flow and out-flow vessels. Acute success was achieved in 100% of the cases without periprocedural complications. Clinical improvement in functional status was obtained and maintained after an average of 12 months, with a significant (P<0.001) decrease in Rutherford class, 100% limb salvage, no major amputation and five (16.1%) minor amputations. Duplex ultrasound control showed restenosis/reocclusion in two (6.5%) cases, whereas a total of seven (22.6%) patients underwent repeat revascularization (2 [6.5%] target lesion re-PTA). CONCLUSIONS: Infra-popliteal PTA with this new, BTK dedicated, long tapered balloon in patients with CLI was feasible and safe, and was associated favorable clinical results at both acute and mid-term follow-up. PMID- 19543197 TI - The effects of calcium dobesilate on the mechanical function of rat hearts. AB - AIM: Calcium dobesilate is an angio-protective agent that has positive effects on hemorheological parameters. It decreases blood and plasma viscosity, thrombocyte aggregation, and microvascular hyperpermeability. It is an antioxidant that increases endothelial-derived vasodilator substance secretion. In this experimental study, the effects of calcium dobesilate on myocardial ischemia reperfusion injury were investigated. METHODS: Using the Langendorff setup, 24 adult Wistar albino rat hearts were perfused. MeanP (mean pressure perfusing the coronary arteries), PSP (maximum left ventricle pressure), +dp/dt(max) (change in contraction power over time), -dp/dt(max) (change in relaxing power over time), PP (peak systolic pressure-minimum balloon pressure) and bpm (number of heart beats per minute) were evaluated. The control group (N.=6) was perfused with Tyrode solution alone. The other three groups (N.=6 for each group) were perfused with the Tyrode solution and calcium dobesilate either before ischemia, during the ischemia reperfusion period, or during the reperfusion-only period. RESULTS: The meanP values were significantly higher in groups perfused by calcium dobesilate. For other parameters, calcium dobesilate did not demonstrate a positive effect. CONCLUSIONS: This study showed that calcium dobesilate may have cardio-protective effects in isolated, perfused rat hearts. In hearts perfused by calcium dobesilate, the increase in mean P may be explained by the increase in endothelium-derived vasodilator substances. Further studies are needed to better characterize the myocardial protective effects of calcium dobesilate. PMID- 19543198 TI - Alcohol and suicide among racial/ethnic populations - 17 states, 2005-2006. AB - During 2001-2005, an estimated annual 79,646 alcohol-attributable deaths (AAD) and 2.3 million years of potential life lost (YPLL) were attributed to the harmful effects of excessive alcohol use. An estimated 5,800 AAD and 189,667 YPLL were associated annually with suicide. The burden of suicide varies widely among racial and ethnic populations in the United States, and limited data are available to describe the role of alcohol in suicides in these populations. To examine the relationship between alcohol and suicide among racial/ethnic populations, CDC analyzed data from the National Violent Death Reporting System (NVDRS) for the 2-year period 2005-2006 (the most recent data available). This report summarizes the results of that analysis, which indicated that the overall prevalence of alcohol intoxication (i.e., blood alcohol concentration [BAC] at or above the legal limit of 0.08 g/dL) was nearly 24% among suicide decedents tested for alcohol, with the highest percentage occurring among American Indian/Alaska Natives (AI/ANs) (37%), followed by Hispanics (29%) and persons aged 20-49 years (28%). These results indicate that many populations can benefit from comprehensive and culturally appropriate suicide-prevention strategies that include efforts to reduce alcohol consumption, especially programs that focus on persons aged <50 years. PMID- 19543199 TI - Novel influenza A (H1N1) virus infections among health-care personnel - United States, April-May 2009. AB - Soon after identification of novel influenza A (H1N1) virus infections in the United States in mid-April 2009, CDC provided interim recommendations to reduce the risk for transmission in health-care settings. These included recommendations on use of personal protective equipment (PPE), management of health-care personnel (HCP) after unprotected exposures, and instruction of ill HCP not to report to work. To better understand the risk for acquiring infection with the virus among HCP and the impact of infection-control recommendations, CDC solicited reports of infected HCP from state health departments. As of May 13, CDC had received 48 reports of confirmed or probable infections with novel influenza A (H1N1) virus; of these, 26 reports included detailed case reports with information regarding risk factors that might have led to infection. Of the 26 cases, 13 (50%) HCP were deemed to have acquired infection in a health-care setting, including one instance of probable HCP to HCP transmission and 12 instances of probable or possible patient to HCP transmission. Eleven HCP had probable or possible acquisition in the community, and two had no reported exposures in either health-care or community settings. Among 11 HCP with probable or possible patient to HCP acquisition and available information on PPE use, only three reported always using either a surgical mask or an N95 respirator. These findings suggest that transmission of novel influenza A (H1N1) virus to HCP is occurring in both health-care and community settings and that additional messages aimed at reinforcing current infection-control recommendations are needed. PMID- 19543200 TI - Large volume (20L+) filtration of coastal seawater samples. AB - The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For this method, samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. This video documents large volume (> or = 20 L) filtration of microbial biomass, ranging between 0.22 microm and 2.7 microm in diameter, from the water column. Two 20 L samples can be filtered simultaneously using a single pump unit equipped with four rotating heads. Filtration is done in the field on extended trips, or immediately upon return for day trips. It is important to record the amount of water passing through each sterivex filter unit. To prevent biofilm formation between sampling trips, all filtration equipment must be rinsed with dilute HCl and deionized water and autoclaved immediately after use. This procedure will take approximately 5 hours plus an additional hour for clean up. PMID- 19543201 TI - Small volume (1-3L) filtration of coastal seawater samples. AB - The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For today's demonstration samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. This video documents small volume (approximately 1 L) filtration of microbial biomass from the water column. The protocol is an extension of the large volume sampling protocol described earlier, with one major difference: here, there is no pre-filtration step, so all size classes of biomass are collected down to the 0.22 microm filter cut-off. Samples collected this way are ideal for nucleic acid analysis. The set-up, filtration, and clean-up steps each take about 20-30 minutes. If using two peristaltic pumps simultaneously, up to 8 samples may be filtered at the same time. To prevent biofilm formation between sampling trips, all filtration equipment must be rinsed with dilute HCl and deionized water and autoclaved immediately after use. PMID- 19543202 TI - FTO genotype is associated with exercise training-induced changes in body composition. AB - The fat mass (FM) and obesity-associated (FTO) gene is the first obesity susceptibility gene identified by genome-wide association scans and confirmed in several follow-up studies. Homozygotes for the risk allele (A/A) have 1.67 times greater risk of obesity than those who do not have the allele. However, it is not known whether regular exercise-induced changes in body composition are influenced by the FTO genotype. The purpose of our study was to test whether the FTO genotype is associated with exercise-induced changes in adiposity. Body composition was derived from underwater weighing before and after a 20-week endurance training program in 481 previously sedentary white subjects of the HERITAGE Family Study. FTO single-nucleotide polymorphism (SNP) rs8050136 was genotyped using Illumina GoldenGate assay. In the sedentary state, the A/A homozygotes were significantly heavier and fatter than the heterozygotes and the C/C homozygotes in men (P = 0.004) but not in women (P = 0.331; gene-by-sex interaction P = 0.0053). The FTO genotype was associated with body fat responses to regular exercise (P < 0.005; adjusted for age, sex, and baseline value of response trait): carriers of the C allele showed three times greater FM and %body fat losses than the A/A homozygotes. The FTO genotype explained 2% of the variance in adiposity changes. Our data suggest that the FTO obesity susceptibility genotype influences the body fat responses to regular exercise. Resistance to exercise-induced reduction in total adiposity may represent one mechanism by which the FTO A allele promotes overweight and obesity. PMID- 19543203 TI - The gene expression of the main lipogenic enzymes is downregulated in visceral adipose tissue of obese subjects. AB - Contradictory findings regarding the gene expression of the main lipogenic enzymes in human adipose tissue depots have been reported. In this cross sectional study, we aimed to evaluate the mRNA expression of fatty acid synthase (FAS) and acetyl-CoA carboxilase (ACC) in omental and subcutaneous (SC) fat depots from subjects who varied widely in terms of body fat mass. FAS and ACC gene expression were evaluated by real time-PCR in 188 samples of visceral adipose tissue which were obtained during elective surgical procedures in 119 women and 69 men. Decreased sex-adjusted FAS (-59%) and ACC (-49%) mRNA were found in visceral adipose tissue from obese subjects, with and without diabetes mellitus type 2 (DM-2), compared with lean subjects (both P < 0.0001). FAS mRNA was also decreased (-40%) in fat depots from overweight subjects (P < 0.05). Indeed, FAS mRNA was significantly and positively associated with ACC gene expression (r = 0.316, P < 0.0001) and negatively with BMI (r = -0.274), waist circumference (r = -0.437), systolic blood pressure (r = -0.310), serum glucose (r = -0.277), and fasting triglycerides (r = -0.226), among others (all P < 0.0001). Similar associations were observed for ACC gene expression levels. In a representative subgroup of nonobese (n = 4) and obese women (n = 6), relative FAS gene expression levels significantly correlated (r = 0.657, P = 0.034; n = 10) with FAS protein values. FAS protein levels were also inversely correlated with blood glucose (r = -0.640, P = 0.046) and fasting triglycerides (r = -0.832, P = 0.010). In conclusion, the gene expression of the main lipogenic enzymes is downregulated in visceral adipose tissue from obese subjects. PMID- 19543204 TI - How many calories are on our plate? Expected fullness, not liking, determines meal-size selection. AB - The availability of highly palatable food is thought to stimulate the selection of larger meals (leading to weight gain and obesity). In this article, we explore aspects of this proposition. Specifically, we scrutinize two basic assumptions: (i) palatable energy-dense foods are more rewarding (desired), and (ii) these palatable foods are selected in relatively larger portions. In combination with palatability, we also consider the relative role for "expected satiation"-the extent to which a food is expected to deliver satiation. A total of 17 commonly consumed foods were assessed by 28 normal-weight participants at lunchtime. Critically, our measure of food reward and expected satiation involves comparisons between foods based on equicaloric portions. When assessed in this way, we find that food reward and ideal portion sizes (in kcal) are both closely associated with expected satiation, but not with "expected liking." Low expected satiation (not expected liking) predicts the selection of large portion sizes (in kcal) and foods with this characteristic tend to be more energy dense and are regarded as less (not more) rewarding (when compared calorie for calorie). Together, these findings challenge the role of palatability in meal-size selection and they highlight the importance of expected satiation, a "nonaffective" component of food reward. PMID- 19543206 TI - BMI and all-cause mortality among Japanese older adults: findings from the Japan collaborative cohort study. AB - The association between BMI and all-cause mortality may vary with gender, age, and ethnic groups. However, few prospective cohort studies have reported the relationship in older Asian populations. We evaluated the association between BMI and all-cause mortality in a cohort comprised 26,747 Japanese subjects aged 65-79 years at baseline (1988-1990). The study participants were followed for an average of 11.2 years. Proportional-hazards regression models were used to estimate mortality hazard ratios (HRs) and 95% confidence intervals. Until 2003, 9,256 deaths occurred. The underweight group was associated with a statistically higher risk of all-cause mortality compared with the mid-normal-range group (BMI: 20.0-22.9); resulting in a 1.78-fold (95% confidence interval: 1.45-2.20) and 2.55-fold (2.13-3.05) increase in mortality risk among severest thin men and women (BMI: <16.0), respectively. Even within the normal-range group, the lower normal-range group (BMI: 18.5-19.9) showed a statistically elevated risk. In contrast, being neither overweight (BMI: 25.0-29.9) nor obese (BMI: > or =30.0) elevated the risk among men; however among women, HR was slightly elevated in the obese group but not in the overweight group compared with the mid-normal-range group. Among Japanese older adults, a low BMI was associated with increased risk of all-cause mortality, even among those with a lower normal BMI range. The wide range of BMI between 20.0 and 29.9 in both older men and women showed the lowest all-cause mortality risk. PMID- 19543205 TI - Acute effects of dietary glycemic index on antioxidant capacity in a nutrient controlled feeding study. AB - Oxidative stress, caused by an imbalance between antioxidant capacity and reactive oxygen species, may be an early event in a metabolic cascade elicited by a high glycemic index (GI) diet, ultimately increasing the risk for cardiovascular disease and diabetes. We conducted a feeding study to evaluate the acute effects of low-GI compared with high-GI diets on oxidative stress and cardiovascular disease risk factors. The crossover study comprised two 10-day in patient admissions to a clinical research center. For the admissions, 12 overweight or obese (BMI: 27-45 kg/m(2)) male subjects aged 18-35 years consumed low-GI or high-GI diets controlled for potentially confounding nutrients. On day 7, after an overnight fast and then during a 5-h postprandial period, we assessed total antioxidant capacity (total and perchloric acid (PCA) protein-precipitated plasma oxygen radical absorbance capacity (ORAC) assay) and oxidative stress status (urinary F(2alpha)-isoprostanes (F(2)IP)). On day 10, we measured cardiovascular disease risk factors. Under fasting conditions, total antioxidant capacity was significantly higher during the low-GI vs. high-GI diet based on total ORAC (11,736 +/- 668 vs. 10,381 +/- 612 micromol Trolox equivalents/l, P = 0.002) and PCA-ORAC (1,276 +/- 96 vs. 1,210 +/- 96 micromol Trolox equivalents/l, P = 0.02). Area under the postprandial response curve also differed significantly between the two diets for total ORAC and PCA-ORAC. No diet effects were observed for the other variables. Enhancement in plasma total antioxidant capacity occurs within 1 week on a low-GI diet, before changes in other risk factors, raising the possibility that this phenomenon may mediate, at least in part, the previously reported effects of GI on health. PMID- 19543207 TI - Role of endogenous ET-1 in the regulation of myocardial blood flow in lean and obese humans. AB - Endothelin is an important determinant of peripheral vascular tone, and increased endogenous endothelin activity contributes to peripheral vascular dysfunction in human obesity. The contributions of endothelin to the regulation of coronary vascular tone in health in humans have not been well studied. We hypothesized that the contribution of endothelin to the regulation of myocardial perfusion would be augmented in human obesity. Using [NH(3)]ammonia positron emission tomography (PET), we measured myocardial perfusion under resting and adenosine stimulated conditions on two separate days, with and without concurrent exposure to BQ123, an antagonist of type A endothelin receptors (1 micromol/min IV beginning 90 min before measurement). We studied 10 lean and 9 obese subjects without hypertension, hyperlipidemia, or diabetes mellitus. We observed a BQ123 induced increase in resting myocardial perfusion of approximately 40%, not different between lean and obese subjects (BQ123-induced increase in flow: lean 0.12 +/- 0.20, obese 0.32 +/- 0.51 ml/g/min, P = 0.02 BQ123 effect, P = 0.27 comparing response across groups). Although basal flow rates varied by region of the myocardium, the BQ123 effect was seen in all regions. BMI and cholesterol were significantly related to BQ123-induced increases in basal tone in multivariable analysis. There was no baseline difference in the adenosine stimulated increase in blood flow between lean and obese subjects, and BQ123 failed to augment these responses in either group. These observations suggest that endothelin is an important contributor to the regulation of myocardial perfusion under resting conditions in healthy lean and obese humans, with increased contributions in proportion to increasing obesity. PMID- 19543208 TI - BMI and mortality: results from a national longitudinal study of Canadian adults. AB - Although a clear risk of mortality is associated with obesity, the risk of mortality associated with overweight is equivocal. The objective of this study is to estimate the relationship between BMI and all-cause mortality in a nationally representative sample of Canadian adults. A sample of 11,326 respondents aged >or=25 in the 1994/1995 National Population Health Survey (Canada) was studied using Cox proportional hazards models. A significant increased risk of mortality over the 12 years of follow-up was observed for underweight (BMI <18.5; relative risk (RR) = 1.73, P < 0.001) and obesity class II+ (BMI >35; RR = 1.36, P <0.05). Overweight (BMI 25 to <30) was associated with a significantly decreased risk of death (RR = 0.83, P < 0.05). The RR was close to one for obesity class I (BMI 30 35; RR = 0.95, P >0.05). Our results are similar to those from other recent studies, confirming that underweight and obesity class II+ are clear risk factors for mortality, and showing that when compared to the acceptable BMI category, overweight appears to be protective against mortality. Obesity class I was not associated with an increased risk of mortality. PMID- 19543209 TI - The associations of LPIN1 gene expression in adipose tissue with metabolic phenotypes in the Chinese population. AB - The LPIN1 gene, encoding lipin-1 protein, plays critical roles in adipocyte differentiation and lipid metabolism. This study aimed to analyze the association of LPIN1 mRNA levels in human adipose tissue with metabolic phenotypes. We also examined the association of LPIN1 genetic variation with type 2 diabetes and related metabolic phenotypes in the Chinese population. The relative LPIN1 mRNA levels were measured in abdominal visceral (VAT) and subcutaneous adipose tissue (SAT) obtained from 102 nondiabetic Chinese females. Seven single-nucleotide polymorphisms (SNPs) spanning from the 5'-upstream region to the 3'-end of the LPIN1 gene were genotyped in 1,520 Chinese (760 type 2 diabetic cases and 760 controls). LPIN1 mRNA levels in VAT were negatively correlated with BMI (r = 0.21, P = 0.03), body fat percentage (r = -0.22, P = 0.02), plasma triglycerides levels (r = -0.21, P = 0.03), and plasma leptin levels (r = -0.63, P = 0.0002). LPIN1 mRNA levels were positively correlated with PPARG and ADIPOQ mRNA levels in both VAT and SAT. No single SNP of the LPIN1 gene was associated with type 2 diabetes in our population. One rare haplotype showed a significant association with type 2 diabetes (odds ratio (OR), 4.35; 95% confidence interval, 1.86-11.75; P = 4 x 10(-4)). No SNP or haplotype of the LPIN1 gene was associated with quantitative metabolic traits in the nondiabetic subjects. The results confirmed the association of LPIN1 gene expression in adipose tissue with lower adiposity and favorable metabolic profiles in the Chinese population. However, the LPIN1 gene seemed not to be a major susceptibility gene for type 2 diabetes or related metabolic phenotypes in the Chinese population. PMID- 19543210 TI - PPARG genotype accounts for part of individual variation in body weight reduction in response to calorie restriction. AB - Several studies indicate that expression of the peroxisome proliferator-activated receptor gamma (PPARG) gene is influenced by calorie restriction. The aim of this study was to investigate whether PPARG gene variations are associated with weight reduction and changes in coronary heart disease (CHD) risk factors in response to a 14-week calorie restriction. In total, 95 middle-aged, Japanese women (BMI>or=25 kg/m2) enrolled as subjects for 14 weeks and attended weekly dietary lectures instructing them on how to consume a nutritionally balanced diet of 1,200 kcal/day. Eight single-nucleotide polymorphisms (SNPs) in the PPARG gene (rs1801282 (Pro/Ala), rs2292101, rs2959272, rs1386835, rs709158, rs1175540, rs1175544, and rs1797912) were analyzed. Body weight decreased significantly ( 7.7+/-3.1 kg; -11.3+/-4.4%) during the intervention. Six PPARG SNPs (rs2959272, rs1386835, rs709158, rs1175540, rs1175544, and rs1797912) were significantly associated with the weight reduction, with rs1175544 having the strongest association (P=0.004). No differences across the rs1175544 genotypes were observed in any of the blood analyses or in blood pressure. In a multiple regression analysis, the rs1175544 genotypes accounted for 7% of the total weight reduction variance. These data suggest that one SNP of the PPARG genotype accounted for a significant portion of the total body weight reduction variance in response to a short-term intervention consisting of calorie restriction; however, no relationship was found between these SNPs and the changes in CHD risk factors which accompanied weight loss. PMID- 19543211 TI - Anandamide increases the differentiation of rat adipocytes and causes PPARgamma and CB1 receptor upregulation. AB - Anandamide (N-arachidonoylethanolamine, AEA) or its metabolites participate in energy balance mainly through feeding modulation. In addition, AEA has been found to increase 3T3-L1 adipocyte differentiation process. In this study, the effect of AEA, R(+)-methanandamide (R(+)-mAEA), URB597, and indomethacin on primary rat adipocyte differentiation was evaluated by a flow cytometry method and by Oil Red O staining. Reverse transcription-PCR and western blotting analysis were performed in order to study the effect of AEA on peroxisome proliferator activated receptor (PPAR)gamma2, cannabinoid receptors (CBRs), fatty acid amidohydrolase (FAAH), and cyclooxygenase-2 (COX-2) expression, during the differentiation process. AEA increased adipocyte differentiation in primary cell cultures in a concentration- and time-dependent manner and induced PPARgamma2 gene expression, confirming findings with 3T3-L1 cell line. CB1R, FAAH, and COX-2 expression was also increased while CB2R expression was decreased. Inhibition of FAAH and COX-2 attenuated the AEA-induced differentiation. Our findings indicate that AEA regulates energy homeostasis not only by appetite modulation but may also regulate adipocyte differentiation and phenotype. PMID- 19543212 TI - Effect of a high-fat diet on 24-hour pattern of circulating adipocytokines in rats. AB - We have shown a significant disruption of 24-h pattern of plasma pituitary, adrenal, and gonadal hormones in high-fat-fed rats. Our objective was to assess the effect of a high-fat diet (35% fat) on mean levels and 24-h pattern of several adipocytokines in rats. A normal diet-fed rats (4% fat) were used as controls. When body weight of high-fat-fed rats attained values about 25% higher than controls (after 66 days of treatment), the animals were killed at six different time intervals throughout a 24-h cycle. Plasma concentrations of insulin, adiponectin, interleukin (IL)-1, leptin, ghrelin, plasminogen activator inhibitor-1 (PAI-1), and monocyte chemoattractant protein-1 (MCP-1) were measured in a multianalyte profiling by using the Luminex-100 system. Tumor necrosis factor alpha (TNFalpha) and IL-6 were measured by enzyme-linked immunosorbent assay. A significant hyperglycemia developed in high-fat-fed rats, together with a significant increase in plasma insulin. Mean levels of plasma adiponectin, IL 1, IL-6, TNFalpha, and leptin augmented, and ghrelin decreased, in high-fat-fed rats. The normal daily pattern of plasma insulin, adiponectin, IL-1, IL-6, TNFalpha, leptin, ghrelin, and MCP-1 became disrupted in high-fat-fed rats. The results indicate that a high-fat diet may bring about signs of insulin resistance and mild inflammation in rats, together with the disruption in daily variations of circulating insulin and ghrelin, and of several adipocytokines including leptin, adiponectin, IL-1, IL-6, TNFalpha, and MCP-1. PMID- 19543213 TI - Cardiac function during exercise in obese prepubertal boys: effect of degree of obesity. AB - The purpose of the study was to evaluate the dynamics of diastolic and systolic function from rest to maximal exercise using conventional echocardiography and tissue Doppler imaging (TDI) in obese prepubertal boys compared to age-matched lean controls. Eighteen obese (10 with first degree obesity and 8 with second degree obesity according to French curves, BMI: 23.3+/-1.8 and 29.0+/-2.0 kg/m2, respectively) and 17 lean controls (BMI=17.6+/-0.6 kg/m2, P<0.001), aged 10-12 years were recruited. After resting echocardiography, all children performed a maximal exercise test. Regional diastolic and systolic myocardial velocities were acquired at rest and each workload. Stroke volume and cardiac output were calculated. At rest, obese boys had greater left ventricular (LV) diameters and LV mass. Boys in the first degree group showed no diastolic or systolic dysfunction, whereas boys with second degree obesity showed subtle diastolic dysfunction. During exercise, both obese groups showed greater stroke volume and cardiac output. First degree obese boys exhibited greater systolic and diastolic tissue Doppler velocities than controls, whereas second degree obese boys had lower diastolic tissue velocities irrespective of exercise intensity and lower fractional shortening at high exercise intensities than controls. In conclusion, no impairment in diastolic or systolic function is noticed in prepubertal boys with first degree of obesity. Enhanced regional myocardial function response to exercise was also demonstrated in this population, suggesting adaptive compensatory cardiac changes in mild obesity. However, when obesity becomes more severe, impaired global and regional cardiac function at rest and during exercise can be observed. PMID- 19543215 TI - Observable weight distributions and children's individual weight assessment. AB - Social networks theory suggests obesity is "contagious" within peer groups in that known friends highly influence weight. On the other hand, an alternative model suggests that observable weight distributions affect perception of one's own obesity level. We examine whether the BMI levels of the most obese classmates in the individual student's grade by gender is positively associated with "under assessment" of obesity and overweight (i.e., independently measured obesity or overweight, but subjective self-assessment of normal weight). The data are the 2004-2005 School Physical Activity and Nutrition III (SPAN), a stratified, multistage probability sample of 4th, 8th, and 11th grade public school children in Texas. We used logistic regression to test whether the gender-specific 85th percentile BMI level within the individual student's grade at their school is positively associated with "under-assessment" of obesity and overweight. The results show that students are much more likely to under-assess their own weight if the gender-specific 85th percentile BMI level is higher in their grade at their school. These data suggest that observable weight distributions play a key role in the obesity epidemic. PMID- 19543214 TI - Influences of normobaric hypoxia training on physical fitness and metabolic risk markers in overweight to obese subjects. AB - Previous studies suggested that hypoxia and exercise may have a synergistic effect on cardiovascular and metabolic risk factors. We conducted a single blind study in overweight to obese subjects to test the hypothesis that training under hypoxia (HG, n = 24, FiO(2) = 15%) results in similar or even greater improvement in body weight and metabolic risk markers compared with exercise under normoxia (NG, n = 21, FiO(2) = 21%). After an initial metabolic evaluation including incremental exercise testing, subjects trained in normoxic or hypoxic conditions thrice weekly over a 4-week period at a heart rate corresponding to 65% of maximum oxygen uptake (VO(2max)). The experimental groups were similar at the start of the investigation and weight stable during the training period. Subjects in the hypoxia group trained at a significantly lower workload (P < 0.05). Yet, both groups showed similar improvements in VO(2max) and time to exhaustion. Respiratory quotient and lactate at the anaerobic threshold as well as body composition improved more in the hypoxia group. We conclude that in obese subjects, training in hypoxia elicits a similar or even better response in terms of physical fitness, metabolic risk markers, and body composition at a lower workload. The fact that workload and, therefore, mechanic strain can be reduced in hypoxia could be particularly beneficial in obese patients with orthopedic comorbidities. PMID- 19543216 TI - Mate tea inhibits in vitro pancreatic lipase activity and has hypolipidemic effect on high-fat diet-induced obese mice. AB - The inhibitory effects of mate tea (MT), a beverage produced with leaves from Ilex paraguariensis, in vitro lipase activity and on obesity in obese mice models were examined. For the in vitro experiment, porcine and human pancreatic lipase (PL) activities were determined by measuring the rate of release of oleic acid from hydrolysis of olive oil emulsified with taurocholate, phospholipids, gum arabic, or polyvinyl alcohol. For the in vivo experiments, animals were fed with a standard diet (SD, n = 10) or high-fat diet (HFD, n = 30) for 16 weeks. After the first 8 weeks on the HFD, the animals were treated with 1 and 2 g/kg of body weight of MT. The time course of the body weight and obesity-related biochemical parameters were evaluated. The results showed that MT inhibited both porcine and human PL (half-maximal inhibitory concentration = 1.5 mg MT/ml) and induced a strong inhibition of the porcine lipase activity in the hydrolysis of substrate emulsified with taurocholate + phosphatidylcholine (PC) (83 +/- 3.8%) or PC alone (62 +/- 4.3%). MT suppressed the increases in body weight (P < 0.05) and decreased the serum triglycerides and low-density lipoprotein (LDL)-cholesterol concentrations at both doses (from 190.3 +/- 5.7 to 135.0 +/- 8.9 mg/dl, from 189.1 +/- 7.3 to 129.3 +/- 17.6 mg/dl; P < 0.05, respectively) after they had been increased by the HFD. The liver lipid content was also decreased by the diet containing MT (from 132.6 +/- 3.9 to 95.6 +/- 6.1 mg/g of tissue; P < 0.05). These results suggest that MT could be a potentially therapeutic alternative in the treatment of obesity caused by a HFD. PMID- 19543217 TI - Interaction of leptin and amylin in the long-term maintenance of weight loss in diet-induced obese rats. AB - We have previously shown that combined amylin + leptin agonism elicits synergistic weight loss in diet-induced obese (DIO) rats. Here, we assessed the comparative efficacy of amylin, leptin, or amylin + leptin in the maintenance of amylin + leptin-mediated weight loss. DIO rats pretreated with the combination of rat amylin (50 microg/kg/day) and murine leptin (125 microg/kg/day) for 4 weeks were subsequently infused with either vehicle, amylin, leptin, or amylin + leptin for an additional 4 weeks. Food intake, body weight, body composition, plasma parameters, and the expression of key metabolic genes in liver and white adipose tissue (WAT) were assessed. Amylin + leptin treatment (weeks 0-4) reduced body weight to 87.5% of baseline. Rats subsequently maintained on vehicle or leptin regained all weight (to 104.2 and 101.2% of baseline, respectively), those maintained on amylin had partial weight regain (97.0%). By contrast, weight loss was largely maintained with continued amylin + leptin treatment (91.4%), associated with a 10% decrease in adiposity. Cumulative food intake (weeks 5-8) was reduced by amylin and amylin + leptin, but not by leptin alone. Amylin + leptin, but not amylin or leptin alone, reduced plasma triglycerides (by 55%), total cholesterol (by 19%), and insulin (by 57%) compared to vehicle. Amylin + leptin also reduced hepatic stearoyl-CoA desaturase-1 (Scd1) mRNA, and increased WAT mRNA levels of adiponectin, fatty acid synthase (Fasn), and lipoprotein lipase (Lpl). We conclude that, in DIO rats, maintenance of amylin + leptin mediated weight loss requires continued treatment with both agonists, and is accompanied by sustained improvements in body composition, and indices of lipid metabolism and insulin sensitivity. PMID- 19543218 TI - Central administration of an endoplasmic reticulum stress inducer inhibits the anorexigenic effects of leptin and insulin. AB - Leptin and insulin are important anorexigenic hormones acting on the hypothalamus. However, most obese humans and animals have reduced hypothalamic responses to leptin and insulin. Increased endoplasmic reticulum (ER) stress has been shown to cause insulin resistance in the livers of obese animals. In the present study, we investigated a role of ER stress in the development of central leptin and insulin resistance. Intracerebroventricular (ICV) administration of the ER stress inducer thapsigargin (TG) increased food intake and body weight. Furthermore, ICV or intra-hypothalamic administration of TG inhibited the anorexigenic and weight-reducing effects of leptin and insulin. ICV administration of TG by itself activated signal-transduction-activated-transcript 3 (STAT3) and Akt in the hypothalamus, but prevented a further activation of hypothalamic STAT3 and Akt by leptin and insulin. We also found that the expression of the ER stress markers such as phosphorylation of the inositol requiring kinase-1 (IRE1), spliced form of X-box-binding protein-1 (XBP-1s), glucose-regulated/binding immunoglobulin protein-78, and C/EBP homology protein (CHOP) increased in the hypothalami of diet-induced obese (DIO) mice. Furthermore, treatment of chemical chaperone 4-phenyl butylic acid significantly improved central leptin resistance in DIO mice. These findings suggest that increased hypothalamic ER stress in obese animals may induce central leptin and insulin resistance. PMID- 19543219 TI - New sites of action for GIRK and SK channels. AB - It was recently discovered that two different types of voltage-insensitive K+ channels, G protein-coupled inwardly rectifying K+ (GIRK) and small-conductance Ca2+-activated K+ (SK) channels, are located on dendritic branches, spines and shafts in the postsynaptic densities of excitatory synapses in many central neurons. Together with increases in our knowledge of how these channels are regulated through stable protein-protein interactions in multi-protein complexes, this has added another layer of complexity to our understanding of synaptic transmission and plasticity. PMID- 19543220 TI - Mitochondrial membrane permeabilization in neuronal injury. AB - Acute neurological conditions such as cerebrovascular diseases and trauma are associated with irreversible loss of neurons and glial cells. Severe or prolonged injury results in uncontrollable cell death within the core of lesions. Conversely, cells that are less severely damaged succumb in a relatively slow fashion, frequently via the intrinsic pathway of cell death, through the deterioration of mitochondrial functions. The permeabilization of mitochondrial membranes determines whether cells will succumb to or survive the injury, and represents a 'point of no return' in mitochondrial cell death. It is therefore an attractive target for the development of new neuroprotective interventions. PMID- 19543222 TI - Principles of neural ensemble physiology underlying the operation of brain machine interfaces. AB - Research on brain-machine interfaces has been ongoing for at least a decade. During this period, simultaneous recordings of the extracellular electrical activity of hundreds of individual neurons have been used for direct, real-time control of various artificial devices. Brain-machine interfaces have also added greatly to our knowledge of the fundamental physiological principles governing the operation of large neural ensembles. Further understanding of these principles is likely to have a key role in the future development of neuroprosthetics for restoring mobility in severely paralysed patients. PMID- 19543223 TI - The 'big pharma' dilemma: develop new drugs or promote existing ones? PMID- 19543221 TI - Circuits controlling vertebrate locomotion: moving in a new direction. AB - Neurobiologists have long sought to understand how circuits in the nervous system are organized to generate the precise neural outputs that underlie particular behaviours. The motor circuits in the spinal cord that control locomotion, commonly referred to as central pattern generator networks, provide an experimentally tractable model system for investigating how moderately complex ensembles of neurons generate select motor behaviours. The advent of novel molecular and genetic techniques coupled with recent advances in our knowledge of spinal cord development means that a comprehensive understanding of how the motor circuitry is organized and operates may be within our grasp. PMID- 19543224 TI - Assessing the translatability of drug projects: what needs to be scored to predict success? AB - Drug development projects have high attrition rates, often because efficacy and safety issues have not been foreseen. More effective prediction of 'translational success' could therefore have a key role in addressing the widely acknowledged problems with weak drug development pipelines. Here, I discuss how a scoring system to systematically assess key determinants of translational success, such as biomarkers and animal and human data, could help identify deficiencies and potential improvements, and increase the reliability of portfolio risk estimates. PMID- 19543226 TI - CD95, BIM and T cell homeostasis. AB - The relative importance of the intrinsic and extrinsic apoptotic pathways in the control of haematopoietic cell homeostasis has been a matter of debate for many years. Cell death is omnipresent in this cellular compartment and ensures the removal of cells that are not properly equipped to assume their function as well as those that have assumed function but are no longer required. In this Review we focus on the roles of CD95 (also known as FAS) and BCL-2-interacting mediator of cell death (BIM), two major regulators of apoptosis in T cell homeostasis, with a particular emphasis on their cooperation in the shutdown of T cell responses. PMID- 19543227 TI - TIP150 interacts with and targets MCAK at the microtubule plus ends. AB - The microtubule (MT) cytoskeleton orchestrates the cellular plasticity and dynamics that underlie morphogenesis and cell division. Growing MT plus ends have emerged as dynamic regulatory machineries in which specialized proteins-called plus-end tracking proteins (+TIPs)-bind to and control the plus-end dynamics that are essential for cell division and migration. However, the molecular mechanisms underlying the plus-end regulation by +TIPs at spindle and astral MTs have remained elusive. Here, we show that TIP150 is a new +TIP that binds to end binding protein 1 (EB1) in vitro and co-localizes with EB1 at the MT plus ends in vivo. Suppression of EB1 eliminates the plus-end localization of TIP150. Interestingly, TIP150 also binds to mitotic centromere-associated kinesin (MCAK), an MT depolymerase that localizes to the plus end of MTs. Suppression of TIP150 diminishes the plus-end localization of MCAK. Importantly, aurora B-mediated phosphorylation disrupts the TIP150-MCAK association in vitro. We reason that TIP150 facilitates the EB1-dependent loading of MCAK onto MT plus ends and orchestrates the dynamics at the plus end of MTs. PMID- 19543228 TI - Futures of ELSA. Science & Society Series on Convergence Research. PMID- 19543229 TI - Water: the invisible problem. Access to fresh water is considered to be a universal and free human right, but dwindling resources and a burgeoning population are increasing its economic value. PMID- 19543225 TI - New insights into the regulation of T cells by gamma(c) family cytokines. AB - Common cytokine receptor gamma-chain (gamma(c)) family cytokines have crucial roles in the development, proliferation, survival and differentiation of multiple cell lineages of both the innate and adaptive immune systems. In this Review, we focus on our current understanding of the distinct and overlapping effects of interleukin-2 (IL-2), IL-7, IL-9, IL-15 and IL-21, as well as the IL-7-related cytokine thymic stromal lymphopoietin (TSLP), on the survival and proliferation of conventional alphabeta T cells, gammadelta T cells and regulatory T cells. This knowledge potentially allows for the therapeutic manipulation of immune responses for the treatment of cancer, autoimmunity, allergic diseases and immunodeficiency, as well as for vaccine development. PMID- 19543230 TI - The role of nuclear pores in gene regulation, development and disease. AB - Nuclear-pore complexes (NPCs) are large protein channels that span the nuclear envelope (NE), which is a double membrane that encloses the nuclear genome of eukaryotes. Each of the typically 2,000-4,000 pores in the NE of vertebrate cells is composed of multiple copies of 30 different proteins known as nucleoporins. The evolutionarily conserved NPC proteins have the well-characterized function of mediating the transport of molecules between the nucleoplasm and the cytoplasm. Mutations in nucleoporins are often linked to specific developmental defects and disease, and the resulting phenotypes are usually interpreted as the consequences of perturbed nuclear transport activity. However, recent evidence suggests that NPCs have additional functions in chromatin organization and gene regulation, some of which might be independent of nuclear transport. Here, we review the transport-dependent and transport-independent roles of NPCs in the regulation of nuclear function and gene expression. PMID- 19543231 TI - Linear polyubiquitination: a new regulator of NF-kappaB activation. AB - The ubiquitin-conjugation system regulates a vast range of biological phenomena by affecting protein function mostly through polyubiquitin conjugation. The type of polyubiquitin chain that is generated seems to determine how conjugated proteins are regulated, as they are recognized specifically by proteins that contain chain-specific ubiquitin-binding motifs. An enzyme complex that catalyses the formation of newly described linear polyubiquitin chains--known as linear ubiquitin chain-assembly complex (LUBAC)--has recently been characterized, as has a particular ubiquitin-binding domain that specifically recognizes linear chains. Both have been shown to have crucial roles in the canonical nuclear factor-kappaB (NF-kappaB)-activation pathway. The ubiquitin system is intimately involved in regulating the NF-kappaB pathway, and the regulatory roles of K63-linked chains have been studied extensively. However, the role of linear chains in this process is only now emerging. This article discusses the possible mechanisms underlying linear polyubiquitin-mediated activation of NF-kappaB, and the different roles that K63-linked and linear chains have in NF-kappaB activation. Future directions for linear polyubiquitin research are also discussed. PMID- 19543232 TI - Induced pluripotent stem cells and the stability of the differentiated state. AB - For much of the last century, the differentiated state that characterizes the many cell types of an adult organism was thought to be stable and abrogated only in rare instances by transdifferentiation, metaplasia or cancer. This stability was thought to reside in the autoregulatory molecular circuitry that exists between the cytoplasm and the nucleus, a status quo that could be disrupted during somatic cell nuclear transfer, to reprogramme cells to a pluripotent state. Pioneering work in the 1980s showed that transdifferentiation of cell lineages could be induced by the addition of transcription factors. However, these conversions were usually confined to cell types from the same germ layer, and proof of conversion was difficult to obtain. This deficiency has now been overturned by demonstrations that exogenously added transcription factors can convert differentiated cell types into embryonic-like induced pluripotent stem cells. Here, we highlight the recent progress, and the implications of this work for our understanding of the relationship between the pluripotent and more differentiated cell states. PMID- 19543233 TI - Genetic basis of hypertension for the development of tailored medicine. AB - Similar to a tsunami wave, a new surge of genome-wide association studies in common complex diseases has succeeded in identifying the causative genetic risk factors of hypertension. The current status of genomic studies in hypertension, however, remains disorganized, and there are no clear solutions in sight. One possible reason for this disorganization is the small effect of each genetic variant on the predisposition to hypertension. Another reason could be that the morbidity of hypertension is typically used as a target to assess genetic contribution, which could be resolved by introducing new technology into the classical genetic analysis. In this review, we reaffirm the genetic basis of hypertension and outline recent progress in genomics. In addition, we discuss both the possibility and usefulness of genomic information in the development of tailored medicine. PMID- 19543234 TI - Combination therapy of angiotensin receptor blocker with statin or thiazolidinediones as promising therapeutic strategy for metabolic syndrome and atherosclerosis. PMID- 19543235 TI - Procaspase 8 overexpression in non-small-cell lung cancer promotes apoptosis induced by FLIP silencing. AB - We found that procaspase 8 was overexpressed in non-small-cell lung cancers (NSCLCs) compared with matched normal tissues. The caspase 8 inhibitor FLICE inhibitory protein (FLIP) was also overexpressed in the majority of NSCLCs. Silencing FLIP induced caspase 8 activation and apoptosis in NSCLC cell lines, but not in normal lung cell lines. Apoptosis induced by FLIP silencing was mediated by the TRAIL death receptors DR4 and DR5, but was not dependent on ligation of the receptors by TRAIL. Furthermore, the apoptosis induced by FLIP silencing was dependent on the overexpression of procaspase 8 in NSCLC cells. Moreover, in NSCLC cells, but not in normal cells, FLIP silencing induced co localization of DR5 and ceramide, and disruption of this co-localization abrogated apoptosis. FLIP silencing supra-additively increased TRAIL-induced apoptosis of NSCLC cells; however, normal lung cells were resistant to TRAIL, even when FLIP was silenced. Importantly, FLIP silencing sensitized NSCLC cells but not normal cells to chemotherapy in vitro, and silencing FLIP in vivo retarded NSCLC xenograft growth and enhanced the anti-tumour effects of cisplatin. Collectively, our results suggest that due to frequent procaspase 8 overexpression, NSCLCs may be particularly sensitive to FLIP-targeted therapies. PMID- 19543237 TI - IL-17 stimulates the proliferation and differentiation of human mesenchymal stem cells: implications for bone remodeling. AB - Interleukin-17 (IL-17) is a cytokine secreted primarily by T(H)-17 cells. Although IL-17 is primarily associated with the induction of tissue inflammation, the other biological roles of IL-17, including non-immune functions, have yet to be thoroughly explored. Here, we report that T-cell-produced IL-17 can induce proliferation of human bone marrow-derived mesenchymal stem cells (hMSCs) in a manner dependent on the generation of reactive oxygen species (ROS). Rac1 GTPase and NADPH oxidase 1 (Nox1) are activated by IL-17 to produce ROS, which in turn stimulates hMSC proliferation. The activation of the MEK-ERK pathway is also crucial for IL-17-dependent hMSC proliferation. TRAF6 and Act1 are required to activate Nox 1 and to phosphorylate MEK on IL-17 stimulation. Interestingly, IL 17 not only accelerates the proliferation of hMSCs, but also induces their migration, motility, and osteoblastic differentiation. Furthermore, IL-17 induces the expression of M-CSF and receptor activator of NF-kappaB ligand (RANKL) on hMSCs, thereby supporting osteoclastogenesis both in vivo and in vitro. On the basis of these results, we suggest that IL-17 can function as a signal to induce extensive bone turnover by regulating hMSC recruitment, proliferation, motility, and differentiation. PMID- 19543236 TI - The multiple levels of regulation by p53 ubiquitination. AB - p53 is a central integrator of a plethora of signals and outputs these signals in the form of tumor suppression. It is well accepted that ubiquitination plays a major part in p53 regulation. Nonetheless, the molecular mechanisms by which p53 activity is controlled by ubiquitination are complex. Mdm2, a RING oncoprotein, was once thought to be the sole E3 ubiquitin ligase for p53, however recent studies have shown that p53 is stabilized but still degraded in the cells of Mdm2 null mice. Although the essential role of Mdm2 in p53 regulation is well established, there are an increasing number of other E3 ligases implicated in Mdm2-independent regulation of p53 by ubiquitination. The different types of ubiquitination on p53 by various E3 ligases have been linked to its differential effects on p53-mediated stress responses. In addition to proteasome-mediated degradation, ubiquitination of p53 acts as signals for degradation-independent functions, such as nuclear export. The function of ubiquitinated p53 varies in the nucleus and cytosol underlying the many potential contributions ubiquitinated p53 may have in promoting cell proliferation or death. Thus, p53 requires multiple layers of regulatory control to ensure correct temporal and spatial functions. PMID- 19543239 TI - Histopathological findings in an epimacular membrane after intraoperative use of perfluorocarbon liquid. PMID- 19543238 TI - Interfering with multimerization of netrin-1 receptors triggers tumor cell death. AB - Netrin-1 was recently proposed to control tumorigenesis by inhibiting apoptosis induced by the dependence receptors DCC (Deleted in colorectal cancer) and UNC5H. Although the loss of these dependence receptors' expression has been described as a selective advantage for tumor growth and progression in numerous cancers, recent observations have shown that some tumors may use an alternative strategy to block dependence receptor-induced programmed cell death: the autocrine expression of netrin-1. This alternative strategy has been observed in a large fraction of aggressive breast cancers, neuroblastoma, pancreatic adenocarcinoma, and lung cancer. This observation is of potential interest regarding future targeted therapy, as in such cases interfering with the ability of netrin-1 to inhibit DCC or UNC5H-induced cell death is associated with apoptosis of netrin-1 expressing tumor cells in vitro, and with inhibition of tumor growth or metastasis in different animal tumor models. The understanding of the mechanism by which netrin-1 inhibits cell death is therefore of interest. Here, we show that netrin-1 triggers the multimerization of both DCC and UNC5H2 receptors, and that multimerization of the intracellular domain of DCC and UNC5H2 is the critical step to inhibit the proapoptotic effects of both of these receptors. Taking advantage of this property, we utilized a recombinant specific domain of DCC that (i) interacts with netrin-1 and (ii) inhibits netrin-1-induced multimerization, to trigger apoptosis in netrin-dependent tumor cells. PMID- 19543240 TI - Bilateral intraocular inflammation after intravitreal bevacizumab in Behcet's disease. PMID- 19543241 TI - Corneal hysteresis in eyes undergoing phototherapeutic keratectomy. PMID- 19543242 TI - Effects of photodynamic therapy on subfoveal blood flow in neovascular age related macular degeneration patients. AB - AIM: To assess the short-term changes in choroidal blood flow (ChBF) after photodynamic therapy (PDT) in patients with neovascular age-related macular degeneration (AMD). METHODS: Fourteen patients with exudative AMD were included after a complete ophthalmological examination, fluorescein and indocyanine green angiography and optical coherence tomography. Subfoveal ChBF was assessed using laser Doppler flowmetry (LDF) in both treated (n=14) and nontreated contralateral (n=8) eyes, 1 h and 1 week after PDT. Ocular perfusion pressure was calculated. RESULTS: The detection sensitivity of the LDF measurements at 2-min intervals before PDT in treated eyes was 7.4% for volume, 6.3% for velocity, and 10.4% for ChBF. The initial mean visual acuity was 0.68+/-0.3 logMar. Macular thickness at baseline as measured by OCT3 was at median (interquartile range), 326.5 microm (188-367). At 1 h and 7 days after PDT, a significant increase in velocity (15.8 and 24.4%, respectively) and a significant decrease in volume (11 and 17.9%, respectively) were noted in treated eyes. Choroidal blood flow and ocular perfusion pressure (OPP) remained similar during follow-up. No significant change in flow parameters was reported in untreated eyes. CONCLUSION: The LDF technique provides feasible and reliable measurements of blood flow parameters before and after PDT in a selective population of patients with exudative AMD. The prognostic value of these early blood flow parameter changes also needs to be assessed. PMID- 19543243 TI - Enhancement of antitumor activity of gammaretrovirus carrying IL-12 gene through genetic modification of envelope targeting HER2 receptor: a promising strategy for bladder cancer therapy. AB - The objective of this study was to develop an HER2-targeted, envelope-modified Moloney murine leukemia virus (MoMLV)-based gammaretroviral vector carrying interleukin (IL)-12 gene for bladder cancer therapy. It displayed a chimeric envelope protein containing a single-chain variable fragment (scFv) antibody to the HER2 receptor and carried the mouse IL-12 gene. The fragment of anti erbB2scFv was constructed into the proline-rich region of the viral envelope of the packaging vector lacking a transmembrane subunit of the carboxyl terminal region of surface subunit. As compared with envelope-unmodified gammaretroviruses, envelope-modified ones had extended viral tropism to human HER2-expressing bladder cancer cell lines, induced apoptosis, and affected cell cycle progression despite lower viral titers. Moreover, animal studies showed that envelope-modified gammaretroviruses carrying IL-12 gene exerted higher antitumor activity in terms of retarding tumor growth and prolonging the survival of tumor-bearing mice than unmodified ones, which were associated with enhanced tumor cell apoptosis as well as increased intratumoral levels of IL-12, interferon-gamma, IL-1beta, and tumor necrosis factor-alpha proteins. Therefore, the antitumor activity of gammaretroviruses carrying the IL-12 gene was enhanced through genetic modification of the envelope targeting HER2 receptor, which may be a promising strategy for bladder cancer therapy. PMID- 19543244 TI - Candidate serous cancer precursors in fallopian tube epithelium of BRCA1/2 mutation carriers. AB - Occult invasive and intraepithelial carcinomas have been identified in the tubal fimbria of BRCA mutation carriers undergoing prophylactic surgery, and recently described lesions overexpressing p53 in the distal tubes of mutation carriers, and non-carriers, have been proposed as histological precursors of high-grade serous carcinoma. The aim of this study was to confirm these findings in a larger, independent case set, to further characterize the cancer precursor lesions, and to determine their frequency in BRCA mutation-positive (n=176) and control groups (n=64). For the purposes of this study, we excluded cases without documentation of a germline mutation of BRCA1/2, and without histological examination of the entire tube, and cases with a diagnosis of invasive carcinoma. Controls included salpingectomies from women undergoing surgery for reasons other than ovarian malignancy. Diagnostic categories were assigned based on combined histological review and immunostaining results. Histological abnormalities were identified in 23% of the BRCA group and in 25% of the control group, and included localized p53 overexpression in 20% of the BRCA group and 25% of the control group. Tubal intramucosal carcinoma was identified in 8% of the BRCA cases and in 3% of the control group. Four cases of intraepithelial carcinoma (21%) did not overexpress p53. There was no significant difference in the median age, frequency of histological abnormalities, p53 signatures, or tubal intraepithelial carcinoma between the BRCA mutation-positive and control groups. This large, blinded review of tubes from BRCA mutation carriers confirms previous reports of putative cancer precursors in distal tubal mucosa, and that p53 signatures occur with similar frequency in women at low and high genetic risk of tubal/ovarian carcinoma. Tubal intraepithelial carcinoma, which, like invasive serous cancer, usually but not always overexpresses p53 protein, is more frequent in BRCA mutation carriers. PMID- 19543245 TI - Adenomatoid tumors of the female and male genital tracts: a clinicopathological and immunohistochemical study of 44 cases. AB - Adenomatoid tumors of the female and male genital tracts are well characterized as mesothelial in origin, but a detailed histological and immunohistochemical analysis comparing both traditional and newer mesothelial markers across gender and site has not been formally conducted. A variety of morphologic features previously described as characteristic of adenomatoid tumors were evaluated in 44 adenomatoid tumors from the male and female genital tracts. Immunohistochemical analysis with pankeratin (AE1/CAM5.2), WT-1, calretinin, CK5/6, D2-40, and caldesmon was also performed. The extent and intensity of staining were scored semiquantitatively on one representative section per case and mean value for each parameter was calculated. All (n=44) the adenomatoid tumors from both the female and male genital tracts demonstrated a distinctive thread-like bridging strand pattern. Lymphoid aggregates were seen in all 12 adenomatoid tumors of male patients, but in only 4 of 32 (13%) tumors in female patients (P<0.0001). The remaining morphologic features were variably present with no clear sex predilection. Pankeratin, calretinin, and D2-40 reactivity were identified in all female (n=32) and male (n=12) genital tract adenomatoid tumors. Adenomatoid tumors expressed WT-1 in 11/12 (92%) male patients and in 31/32 (97%) female patients. In male patients, reactivity for CK5/6 and caldesmon was found in 1/12 (8%) and 0/12 (0%) adenomatoid tumors (respectively), whereas reactivity in female patients was found in 5/32 (16%) and 1/32 (3%); respectively. Female tumors differ from their male counterparts by the frequent absence of lymphoid aggregates and the presence of a circumscribed margin when occurring in the fallopian tube. Of the putative mesothelial markers evaluated, calretinin, D2-40, and WT-1 show a similar immunoprofile and have a higher sensitivity than CK5/6 and caldesmon in genital tract adenomatoid tumors. However, the presence of additional, often strong expression of WT-1 in normal tissues of the female genital tract limits the utility of WT-1 in this setting. PMID- 19543246 TI - Papillary carcinoma of the breast lacks evidence of RET rearrangements despite morphological similarities to papillary thyroid carcinoma. AB - Rare breast neoplasms resembling the tall-cell variant of papillary thyroid carcinoma have been reported. In addition, papillary carcinoma of the breast occasionally displays nuclear features reminiscent of papillary thyroid carcinoma. In this study, we evaluated 33 intraductal/intracystic papillary carcinomas of the breast for the presence and extent of nuclear overlap, grooves, clearing, and inclusions, as well as features of the tall-cell or columnar-cell variants of papillary thyroid carcinoma. RET rearrangements were assessed in a subset of these cases. Paired probes localizing to the centromeric and telomeric ends of the RET gene on chromosome 10 were used for FISH using a break-apart approach. Single round and nested PCR was performed to detect RET/PTC1, RET/PTC2, RET/PTC3 and ELKS-RET fusion transcripts. Nuclear overlap, grooves, stratification, and clearing were identified in 24 (73%), 14 (42%), 11 (33%), and 9 (27%) cases respectively, whereas nuclear inclusions and 'tall-cell' features were each seen in only one (3%) and two (6%) cases, respectively. Four of 19 tested cases displayed split FISH signals in a low percentage of cells and were considered borderline for RET rearrangement. All 19 tested cases with amplifiable RNA were negative for the four RET fusion transcripts evaluated by RT-PCR. Although papillary carcinomas of breast often display one or more cytoarchitectural features of papillary thyroid carcinoma, there is no evidence that RET rearrangements are involved. PMID- 19543247 TI - Chromosomal copy number changes supporting the classification of lentiginous junctional melanoma of the elderly as a subtype of melanoma. AB - Recently the term lentiginous melanoma of the elderly has been suggested for a pattern of melanocytic neoplasia characterized by frequent occurrence in elderly patients, broad lentiginous growth pattern, with poorly cohesive nesting, suprabasilar extension of melanocytes and moderate cytological atypia. However, there are limited reported cases with follow-up information to confirm the malignant nature of these neoplasms. Using fluorescence in situ hybridization (FISH) targeting chromosomal loci that are frequently found to have copy number changes in melanoma, we evaluated cases of lentiginous junctional melanoma of the elderly in order to compare with the frequencies and patterns of chromosomal aberrations identified in other subtypes of melanoma. Previous studies have shown that by using a FISH assay targeting 6p25, 6q23, 11q13 and CEP6 with previously validated criteria, one could discriminate benign nevi from melanoma with high sensitivity and specificity. In this study, 16 of 19 cases (84%) showed sufficient copy number changes in one of the targeted chromosomal loci to meet FISH criteria for melanoma. A total of 17 control cases of lentiginous junctional nevi tested negative for all criteria. These findings support the classification of this pattern of melanocytic neoplasia as a subtype of melanoma. PMID- 19543253 TI - Required modifications and designated member reviews. Expedite the process. PMID- 19543254 TI - Required modifications and designated member reviews. PMID- 19543256 TI - Required modifications and designated member reviews. A time-saver. PMID- 19543255 TI - Required modifications and designated member reviews. Not a shortcut. PMID- 19543257 TI - Required modifications and designated member reviews. A word from OLAW and USDA. PMID- 19543258 TI - Blistering dermatosis in a Yucatan minipig. Spontaneous bullous pemphigoid. PMID- 19543260 TI - Canine and feline tail amputation. PMID- 19543261 TI - Comparison of ketamine-pentobarbital anesthesia and fentanyl-pentobarbital anesthesia for open-heart surgery in minipigs. AB - The authors analyzed and compared the cardiovascular effects of two anesthetic combinations in minipigs undergoing open-heart surgery and cardiopulmonary bypass. Pigs in group K (n = 15) were anesthetized with low-dose ketamine combined with pentobarbital (5 mg per kg and 20 mg per kg, respectively, for induction; continuous intravenous infusion of 5 mg per kg per h and 10 mg per kg per h, respectively, for maintenance). Pigs in group F (n = 15) were treated with fentanyl and pentobarbital (20 microg per kg and 20 mg per kg, respectively, for induction; continuous intravenous infusion of 20 microg per kg per h and 10 mg per kg per h, respectively, for maintenance). Most pigs remained stable during the surgical procedures and survived for at least one day after surgery; two pigs in group F died during or soon after surgery. Heart rate and mean arterial pressure after bypass were significantly lower in group F than in group K, and pigs in group F required higher doses of inotropic agents to maintain cardiac function. Results suggest that for open-heart surgery and bypass in minipigs, ketamine-pentobarbital anesthesia is associated with more stable cardiovascular conditions than is fentanyl-pentobarbital anesthesia. PMID- 19543262 TI - Sonographically guided placement of intravenous catheters in minipigs. AB - Many procedures in minipigs require establishment of reliable deep venous access with a large-bore catheter. In animal experiments, such catheters are typically implanted surgically. In clinical settings, however, ultrasound imaging is routinely used to facilitate safe, minimally invasive puncture of deep vessels. The authors describe a technique for using ultrasound guidance to puncture and cannulate the minipig femoral vein. They carried out the procedure in six minipigs for the purpose of injecting contrast agents for subsequent imaging scans. The procedure was ultimately successful in all pigs, took 10 min on average and resulted in no physiological complications. In one minipig, however, a 10-cm-long catheter became dislodged from the femoral vein; use of a longer (25 cm-long) catheter was optimal for establishing reliable intravenous access. PMID- 19543263 TI - A simple method for assessing analgesic requirements and efficacy in rodents. AB - Evaluation of pain in the clinical setting is an ongoing challenge for veterinarians, researchers and IACUCs. Behavioral assessment, a common technique for evaluating pain, is subjective and difficult to translate into quantifiable data. The authors propose measuring changes in body weight, food consumption and water consumption as a simple and objective method for evaluating postsurgical pain and analgesic efficacy in rodents. PMID- 19543264 TI - An officer and a businessman. PMID- 19543265 TI - The CREB coactivator CRTC2 links hepatic ER stress and fasting gluconeogenesis. AB - In fasted mammals, circulating pancreatic glucagon stimulates hepatic gluconeogenesis in part through the CREB regulated transcription coactivator 2 (CRTC2, also referred to as TORC2). Hepatic glucose production is increased in obesity, reflecting chronic increases in endoplasmic reticulum (ER) stress that promote insulin resistance. Whether ER stress also modulates the gluconeogenic program directly, however, is unclear. Here we show that CRTC2 functions as a dual sensor for ER stress and fasting signals. Acute increases in ER stress triggered the dephosphorylation and nuclear entry of CRTC2, which in turn promoted the expression of ER quality control genes through an association with activating transcription factor 6 alpha (ATF6alpha, also known as ATF6)--an integral branch of the unfolded protein response. In addition to mediating CRTC2 recruitment to ER stress inducible promoters, ATF6alpha also reduced hepatic glucose output by disrupting the CREB-CRTC2 interaction and thereby inhibiting CRTC2 occupancy over gluconeogenic genes. Conversely, hepatic glucose output was upregulated when hepatic ATF6alpha protein amounts were reduced, either by RNA interference (RNAi)-mediated knockdown or as a result of persistent stress in obesity. Because ATF6alpha overexpression in the livers of obese mice reversed CRTC2 effects on the gluconeogenic program and lowered hepatic glucose output, our results demonstrate how cross-talk between ER stress and fasting pathways at the level of a transcriptional coactivator contributes to glucose homeostasis. PMID- 19543266 TI - mTOR regulates memory CD8 T-cell differentiation. AB - Memory CD8 T cells are a critical component of protective immunity, and inducing effective memory T-cell responses is a major goal of vaccines against chronic infections and tumours. Considerable effort has gone into designing vaccine regimens that will increase the magnitude of the memory response, but there has been minimal emphasis on developing strategies to improve the functional qualities of memory T cells. Here we show that mTOR (mammalian target of rapamycin, also known as FRAP1) is a major regulator of memory CD8 T-cell differentiation, and in contrast to what we expected, the immunosuppressive drug rapamycin has immunostimulatory effects on the generation of memory CD8 T cells. Treatment of mice with rapamycin following acute lymphocytic choriomeningitis virus infection enhanced not only the quantity but also the quality of virus specific CD8 T cells. Similar effects were seen after immunization of mice with a vaccine based on non-replicating virus-like particles. In addition, rapamycin treatment also enhanced memory T-cell responses in non-human primates following vaccination with modified vaccinia virus Ankara. Rapamycin was effective during both the expansion and contraction phases of the T-cell response; during the expansion phase it increased the number of memory precursors, and during the contraction phase (effector to memory transition) it accelerated the memory T cell differentiation program. Experiments using RNA interference to inhibit expression of mTOR, raptor (also known as 4932417H02Rik) or FKBP12 (also known as FKBP1A) in antigen-specific CD8 T cells showed that mTOR acts intrinsically through the mTORC1 (mTOR complex 1) pathway to regulate memory T-cell differentiation. Thus these studies identify a molecular pathway regulating memory formation and provide an effective strategy for improving the functional qualities of vaccine- or infection-induced memory T cells. PMID- 19543267 TI - The establishment of gene silencing at single-cell resolution. AB - The establishment of silencing in Saccharomyces cerevisiae is similar to heterochromatin formation in multicellular eukaryotes. Previous batch culture studies determined that the de novo establishment of silencing initiates during S phase and continues for up to five cell divisions for completion. To track silencing phenotypically, we developed an assay that introduces Sir3 protein into individual sir3Delta mutant cells synchronously and then detects the onset of silencing with single-cell resolution. Silencing was completed within the first one to two cell divisions in most cells queried. Moreover, we uncovered unexpected complexity in the contributions of a histone acetyltransferase (Sas2), two histone methytransferases (Dot1 and Set1) and one histone demethylase (Jhd2) to the dynamics of silencing. Our findings showed that removal of methyl modifications at H3K4 and H3K79 were important steps in silent chromatin formation and that Jhd2 and Set1 had competing roles in the process. PMID- 19543268 TI - Wnt9b signaling regulates planar cell polarity and kidney tubule morphogenesis. AB - Although many vertebrate organs, such as kidneys, lungs and liver, are composed of epithelial tubules, little is known of the mechanisms that establish the length or diameter of these tubules. In the kidney, defects in the establishment or maintenance of tubule diameter are associated with one of the most common inherited human disorders, polycystic kidney disease. Here we show that attenuation of Wnt9b signaling during kidney morphogenesis affects the planar cell polarity of the epithelium and leads to tubules with significantly increased diameter. Although previous studies showed that polarized cell divisions maintain the diameter of postnatal kidney tubules, we find that cell divisions are randomly oriented during embryonic development. Our data suggest that diameter is established during early morphogenetic stages by convergent extension processes and maintained by polarized cell divisions. Wnt9b, signaling through the non canonical Rho/Jnk branch of the Wnt pathway, is necessary for both of these processes. PMID- 19543269 TI - The DNA replication FoSTeS/MMBIR mechanism can generate genomic, genic and exonic complex rearrangements in humans. AB - We recently proposed a DNA replication-based mechanism of fork stalling and template switching (FoSTeS) to explain the complex genomic rearrangements associated with a dysmyelinating central nervous system disorder in humans. The FoSTeS mechanism has been further generalized and molecular mechanistic details have been provided in the microhomology-mediated break-induced replication (MMBIR) model that may underlie many structural variations in genomes from all domains of life. Here we provide evidence that human genomic rearrangements ranging in size from several megabases to a few hundred base pairs can be generated by FoSTeS/MMBIR. Furthermore, we show that FoSTeS/MMBIR-mediated rearrangements can occur mitotically and can result in duplication or triplication of individual genes or even rearrangements of single exons. The FoSTeS/MMBIR mechanism can explain both the gene duplication-divergence hypothesis and exon shuffling, suggesting an important role in both genome and single-gene evolution. PMID- 19543270 TI - Centromere assembly requires the direct recognition of CENP-A nucleosomes by CENP N. AB - Centromeres are specialized chromosomal domains that direct kinetochore assembly during mitosis. CENP-A (centromere protein A), a histone H3-variant present exclusively in centromeric nucleosomes, is thought to function as an epigenetic mark that specifies centromere identity. Here we identify the essential centromere protein CENP-N as the first protein to selectively bind CENP-A nucleosomes but not H3 nucleosomes. CENP-N bound CENP-A nucleosomes in a DNA sequence-independent manner, but did not bind soluble CENP-A-H4 tetramers. Mutations in CENP-N that reduced its affinity for CENP-A nucleosomes caused defects in CENP-N localization and had dominant effects on the recruitment of CENP-H, CENP-I and CENP-K to centromeres. Depletion of CENP-N using siRNA (short interfering RNA) led to similar centromere assembly defects and resulted in reduced assembly of nascent CENP-A into centromeric chromatin. These data suggest that CENP-N interprets the information encoded within CENP-A nucleosomes and recruits other proteins to centromeric chromatin that are required for centromere function and propagation. PMID- 19543272 TI - Increasing organismal healthspan by enhancing mitochondrial protein quality control. AB - Degradation of damaged proteins by members of the protein quality control system is of fundamental importance in maintaining cellular homeostasis. In mitochondria, organelles which both generate and are targets of reactive oxygen species (ROS), a number of membrane bound and soluble proteases are essential components of this system. Here we describe the regulation of Podospora anserina LON (PaLON) levels, an AAA(+) family serine protease localized in the matrix fraction of mitochondria. Constitutive overexpression of PaLon results in transgenic strains of the fungal ageing model P. anserina showing increased ATP dependent serine protease activity. These strains display lower levels of carbonylated (aconitase) and carboxymethylated proteins, reduced secretion of hydrogen peroxide and a higher resistance against exogenous oxidative stress. Moreover, they are characterized by an extended lifespan without impairment of vital functions such as respiration, growth and fertility. The reported genetic manipulation proved to be a successful intervention in organismal ageing and it led to an increase in the healthy lifespan, the healthspan, of P. anserina. PMID- 19543271 TI - TGF-beta activates Akt kinase through a microRNA-dependent amplifying circuit targeting PTEN. AB - Akt kinase is activated by transforming growth factor-beta1 (TGF-beta) in diabetic kidneys, and has important roles in fibrosis, hypertrophy and cell survival in glomerular mesangial cells. However, the mechanisms of Akt activation by TGF-beta are not fully understood. Here we show that TGF-beta activates Akt in glomerular mesangial cells by inducing the microRNAs (miRNAs) miR-216a and miR 217, both of which target PTEN (phosphatase and tensin homologue), an inhibitor of Akt activation. These miRNAs are located within the second intron of a non coding RNA (RP23-298H6.1-001). The RP23 promoter was activated by TGF-beta and miR-192 through E-box-regulated mechanisms, as shown previously. Akt activation by these miRs led to glomerular mesangial cell survival and hypertrophy, which were similar to the effects of activation by TGF-beta. These studies reveal a mechanism of Akt activation through PTEN downregulation by two miRs, which are regulated by upstream miR-192 and TGF-beta. Due to the diversity of PTEN function, this miR-amplifying circuit may have key roles, not only in kidney disorders, but also in other diseases. PMID- 19543273 TI - OsHAL3 mediates a new pathway in the light-regulated growth of rice. AB - Plants show distinct morphologies in different light conditions through a process called photomorphogenesis. A predominant feature of photomorphogenesis is the reduced growth of seedlings under light conditions compared with darkness. For this adaptive event, the most well-known molecular mechanism involves photoreceptor-mediated inhibition of cell elongation. However, it is not known whether additional pathways exist. Here, we describe a newly discovered pathway of light-modulated plant growth mediated by the halotolerance protein HAL3, a flavin mononucleotide (FMN)-binding protein involved in cell division. We found that light, especially blue light, suppresses growth of rice seedlings by reducing the activity of Oryza sativa (Os) HAL3. Both in vitro and in vivo studies showed that OsHAL3 is structurally inactivated by light through photo oxidation and by direct interaction with photons. In addition, the transcriptional expression of OsHAL3 is synergistically regulated by different light conditions. Further investigation suggested that OsHAL3 promotes cell division by recruiting a ubiquitin system, rather than by its 4' phosphopantothenoylcysteine (PPC) decarboxylase activity. Our results uncover a new mechanism for light-regulated plant growth, namely, light not only inhibits cell elongation but also suppresses cell division through HAL3 and E3 ubiquitin ligase. This study thus brings new insights into our understanding of plant photomorphogenesis. PMID- 19543276 TI - Linking a completely three-dimensional nanostrain to a structural transformation eigenstrain. AB - Ni-Ti is one of the most popular shape-memory alloys, a phenomenon resulting from a martensitic transformation. Commercial Ni-Ti-based alloys are often thermally treated to contain Ni(4)Ti(3) precipitates. The presence of these precipitates can introduce an extra transformation step related to the so-called R-phase. It is believed that the strain field surrounding the precipitates, caused by the matrix-precipitate lattice mismatch, lies at the origin of this intermediate transformation step. Atomic-resolution transmission electron microscopy in combination with geometrical phase analysis is used to measure the elastic strain field surrounding these precipitates. By combining measurements from two different crystallographic directions, the three-dimensional strain matrix is determined from two-dimensional measurements. Comparison of the measured strain matrix to the eigenstrain of the R-phase shows that both are very similar and that the introduction of the R-phase might indeed compensate the elastic strain introduced by the precipitate. PMID- 19543275 TI - The impact of negative selection on thymocyte migration in the medulla. AB - Developing thymocytes are screened for self-reactivity before they exit the thymus, but how thymocytes scan the medulla for self antigens is unclear. Using two-photon microscopy, we observed that medullary thymocytes migrated rapidly and made frequent, transient contacts with dendritic cells. In the presence of a negative selecting ligand, thymocytes slowed, became confined to areas of approximately 30 microm in diameter and had increased contact with dendritic cells surrounding confinement zones. One third of polyclonal medullary thymocytes also showed confined, slower migration and may correspond to autoreactive thymocytes. Our data suggest that many autoreactive thymocytes do not undergo immediate arrest and death after encountering a negative selecting ligand but instead adopt an altered migration program while remaining in the medullary microenvironment. PMID- 19543274 TI - The Arp2/3 complex and WASp are required for apical trafficking of Delta into microvilli during cell fate specification of sensory organ precursors. AB - Cell fate decisions mediated by the Notch signalling pathway require direct cell cell contact between adjacent cells. In Drosophila melanogaster, an external sensory organ (ESO) develops from a single sensory organ precursor (SOP) and its fate specification is governed by differential Notch activation. Here we show that mutations in actin-related protein-3 (Arp3) compromise Notch signalling, leading to a fate transformation of the ESO. Our data reveal that during ESO fate specification, most endocytosed vesicles containing the ligand Delta traffic to a prominent apical actin-rich structure (ARS) formed in the SOP daughter cells. Using immunohistochemistry and transmission electron microscopy (TEM) analyses, we show that the ARS contains numerous microvilli on the apical surface of SOP progeny. In Arp2/3 and WASp mutants, the surface area of the ARS is substantially reduced and there are significantly fewer microvilli. More importantly, trafficking of Delta-positive vesicles from the basal area to the apical portion of the ARS is severely compromised. Our data indicate that WASp-dependent Arp2/3 actin polymerization is crucial for apical presentation of Delta, providing a mechanistic link between actin polymerization and Notch signalling. PMID- 19543277 TI - Atomic-scale imaging of individual dopant atoms in a buried interface. AB - Determining the atomic structure of internal interfaces in materials and devices is critical to understanding their functional properties. Interfacial doping is one promising technique for controlling interfacial properties at the atomic scale, but it is still a major challenge to directly characterize individual dopant atoms within buried crystalline interfaces. Here, we demonstrate atomic scale plan-view observation of a buried crystalline interface (an yttrium-doped alumina high-angle grain boundary) using aberration-corrected Z-contrast scanning transmission electron microscopy. The focused electron beam transmitted through the off-axis crystals clearly highlights the individual yttrium atoms located on the monoatomic layer interface plane. Not only is their unique two-dimensional ordered positioning directly revealed with atomic precision, but local disordering at the single-atom level, which has never been detected by the conventional approaches, is also uncovered. The ability to directly probe individual atoms within buried interface structures adds new dimensions to the atomic-scale characterization of internal interfaces and other defect structures in many advanced materials and devices. PMID- 19543278 TI - The dynamic organic p-n junction. AB - Static p-n junctions in inorganic semiconductors are exploited in a wide range of today's electronic appliances. Here, we demonstrate the in situ formation of a dynamic p-n junction structure within an organic semiconductor through electrochemistry. Specifically, we use scanning kelvin probe microscopy and optical probing on planar light-emitting electrochemical cells (LECs) with a mixture of a conjugated polymer and an electrolyte connecting two electrodes separated by 120 microm. We find that a significant portion of the potential drop between the electrodes coincides with the location of a thin and distinct light emission zone positioned >30 microm away from the negative electrode. These results are relevant in the context of a long-standing scientific debate, as they prove that electrochemical doping can take place in LECs. Moreover, a study on the doping formation and dissipation kinetics provides interesting detail regarding the electronic structure and stability of the dynamic organic p-n junction, which may be useful in future dynamic p-n junction-based devices. PMID- 19543279 TI - Sequential click reactions for synthesizing and patterning three-dimensional cell microenvironments. AB - Click chemistry provides extremely selective and orthogonal reactions that proceed with high efficiency and under a variety of mild conditions, the most common example being the copper(I)-catalysed reaction of azides with alkynes. While the versatility of click reactions has been broadly exploited, a major limitation is the intrinsic toxicity of the synthetic schemes and the inability to translate these approaches into biological applications. This manuscript introduces a robust synthetic strategy where macromolecular precursors react through a copper-free click chemistry, allowing for the direct encapsulation of cells within click hydrogels for the first time. Subsequently, an orthogonal thiol-ene photocoupling chemistry is introduced that enables patterning of biological functionalities within the gel in real time and with micrometre-scale resolution. This material system enables us to tailor independently the biophysical and biochemical properties of the cell culture microenvironments in situ. This synthetic approach uniquely allows for the direct fabrication of biologically functionalized gels with ideal structures that can be photopatterned, and all in the presence of cells. PMID- 19543280 TI - Cellular and systems mechanisms of memory strength as a constraint on auditory fear reconsolidation. AB - Memory reconsolidation has been demonstrated in various tasks and species, suggesting it is a fundamental process. However, there are experimental parameters that can inhibit reconsolidation from occurring (boundary conditions). These conditions and their mechanisms remain poorly defined. Here, we characterize the ability of strong training to inhibit reconsolidation at the behavioral, systems and molecular levels. We demonstrate that strong memories in rats initially are resistant to reconsolidation, but after sufficient time will undergo reconsolidation, suggesting that boundary conditions can be transient. At the systems level, we show that the hippocampus is necessary for inhibiting reconsolidation in the amygdala. At the molecular level, we demonstrate that NR2B NMDA-receptor subunits which are critical for the induction of reconsolidation of auditory memories in the amygdala, are downregulated only under conditions when strong memories do not undergo reconsolidation. This suggests that one molecular mechanism for mediating boundary conditions is through downregulation of reconsolidation induction mechanisms. PMID- 19543281 TI - Roles of stargazin and phosphorylation in the control of AMPA receptor subcellular distribution. AB - Understanding how the subcellular fate of newly synthesized AMPA receptors (AMPARs) is controlled is important for elucidating the mechanisms of neuronal function. We examined the effect of increased synthesis of AMPAR subunits on their subcellular distribution in rat hippocampal neurons. Virally expressed AMPAR subunits (GluR1 or GluR2) accumulated in cell bodies and replaced endogenous dendritic AMPAR with little effect on total dendritic amounts and caused no change in synaptic transmission. Coexpressing stargazin (STG) or mimicking GluR1 phosphorylation enhanced dendritic GluR1 levels by protecting GluR1 from lysosomal degradation. However, STG interaction or GluR1 phosphorylation did not increase surface or synaptic GluR1 levels. Unlike GluR1, STG did not protect GluR2 from lysosomal degradation or increase dendritic GluR2 levels. In general, AMPAR surface levels, and not intracellular amounts, correlated strongly with synaptic levels. Our results suggest that AMPAR surface expression, but not its intracellular production or accumulation, is critical for regulating synaptic transmission. PMID- 19543282 TI - Scavenger receptor B2 is a cellular receptor for enterovirus 71. AB - Enterovirus 71 (EV71) belongs to human enterovirus species A of the genus Enterovirus within the family Picornaviridae. EV71, together with coxsackievirus A16 (CVA16), are most frequently associated with hand, foot and mouth disease (HFMD). Although HFMD is considered a mild exanthematous infection, infections involving EV71, but not CVA16, can progress to severe neurological disease, including fatal encephalitis, aseptic meningitis and acute flaccid paralysis. In recent years, epidemic and sporadic outbreaks of neurovirulent EV71 infections have been reported in Taiwan, Malaysia, Singapore, Japan and China. Here, we show that human scavenger receptor class B, member 2 (SCARB2, also known as lysosomal integral membrane protein II or CD36b like-2) is a receptor for EV71. EV71 binds soluble SCARB2 or cells expressing SCARB2, and the binding is inhibited by an antibody to SCARB2. Expression of human SCARB2 enables normally unsusceptible cell lines to support EV71 propagation and develop cytopathic effects. EV71 infection is hampered by the antibody to SCARB2 and soluble SCARB2. SCARB2 also supports the infection of the milder pathogen CVA16. The identification of SCARB2 as an EV71 and CVA16 receptor contributes to a better understanding of the pathogenicity of these viruses. PMID- 19543284 TI - Human P-selectin glycoprotein ligand-1 is a functional receptor for enterovirus 71. AB - Enterovirus 71 (EV71) is a major causative agent of hand, foot and mouth disease (HFMD), a common febrile disease occurring mainly in young children. Although clinical manifestations of HFMD are usually mild and self limiting, a severe EV71 outbreak can lead to a diverse array of neurological diseases. Identification of the specific cellular receptors is crucial for elucidating the mechanism of early virus-host interactions and the pathogenesis of enteroviruses. Here we identify human P-selectin glycoprotein ligand-1 (PSGL-1; CD162), a sialomucin membrane protein expressed on leukocytes that has a major role in early stages of inflammation, as a functional receptor for EV71 using an expression cloning method by panning. The N-terminal region of PSGL-1 binds specifically to EV71. Stable PSGL-1 expression allowed EV71 entry and replication, and development of cytopathic effects in nonsusceptible mouse L929 cells. Five out of eight EV71 strains bound soluble PSGL-1 and used intact PSGL-1 as the primary receptor for infection of Jurkat T cells. Three other EV71 strains did not use PSGL-1, suggesting the presence of strain-specific replication of EV71 in leukocytes. EV71 replicated in nonleukocyte cell lines in a PSGL-1-independent manner, indicating the presence of alternative receptor(s) for EV71. The identification of PSGL-1 as a receptor for EV71 sheds new light on a role for PSGL-1-positive leukocytes in cell tropism and pathogenesis during the course of HFMD and other EV71-mediated diseases. PMID- 19543283 TI - HIV reservoir size and persistence are driven by T cell survival and homeostatic proliferation. AB - HIV persists in a reservoir of latently infected CD4(+) T cells in individuals treated with highly active antiretroviral therapy (HAART). Here we identify central memory (T(CM)) and transitional memory (T(TM)) CD4(+) T cells as the major cellular reservoirs for HIV and find that viral persistence is ensured by two different mechanisms. HIV primarily persists in T(CM) cells in subjects showing reconstitution of the CD4(+) compartment upon HAART. This reservoir is maintained through T cell survival and low-level antigen-driven proliferation and is slowly depleted with time. In contrast, proviral DNA is preferentially detected in T(TM) cells from aviremic individuals with low CD4(+) counts and higher amounts of interleukin-7-mediated homeostatic proliferation, a mechanism that ensures the persistence of these cells. Our results suggest that viral eradication might be achieved through the combined use of strategic interventions targeting viral replication and, as in cancer, drugs that interfere with the self renewal and persistence of proliferating memory T cells. PMID- 19543285 TI - Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers. AB - The use of fluorescently labeled major histocompatibility complex multimers has become an essential technique for analyzing disease- and therapy-induced T-cell immunity. Whereas classical major histocompatibility complex multimer analyses are well-suited for the detection of immune responses to a few epitopes, limitations on human-subject sample size preclude a comprehensive analysis of T cell immunity. To address this issue, we developed a combinatorial encoding strategy that allows the parallel detection of a multitude of different T-cell populations in a single sample. Detection of T cells from peripheral blood by combinatorial encoding is as efficient as detection with conventionally labeled multimers but results in a substantially increased sensitivity and, most notably, allows comprehensive screens to be performed. We obtained proof of principle for the feasibility of large-scale screening of human material by analysis of human leukocyte antigen A3-restricted T-cell responses to known and potential melanoma associated antigens in peripheral blood from individuals with melanoma. PMID- 19543286 TI - Simultaneous detection of many T-cell specificities using combinatorial tetramer staining. AB - The direct detection of antigen-specific T cells using tetramers of soluble peptide-major histocompatibilty complex (pMHC) molecules is widely used in both basic and clinical immunology. However, the number of specificities that can be assessed simultaneously has been a major limitation. Here we describe and validate a method using combinations of fluorescent pMHC tetramers to simultaneously detect and enrich for many (>or=15) T-cell specificities in a single human blood sample. PMID- 19543287 TI - Filter-based hybridization capture of subgenomes enables resequencing and copy number detection. AB - To exploit contemporary sequencing technologies for targeted genetic analyses, we developed a hybridization enrichment strategy for DNA capture that uses PCR products as subgenomic traps. We applied this strategy to 115 kilobases of the human genome encompassing 47 genes implicated in cardiovascular disease. Massively parallel sequencing of captured subgenomic libraries interrogated 99.8% of targeted nucleotides >or=20 times ( approximately 40,000-fold enrichment), enabling sensitive and specific detection of sequence variation and copy-number variation. PMID- 19543288 TI - Mode of VAMP substrate recognition and inhibition of Clostridium botulinum neurotoxin F. AB - Clostridium botulinum neurotoxins (BoNTs) cleave neuronal proteins responsible for neurotransmitter release, causing the neuroparalytic disease botulism. BoNT serotypes B, D, F and G cleave and inactivate vesicle-associated membrane protein (VAMP), each at a unique peptide bond. The specificity of BoNTs depends on the mode of substrate recognition. We have investigated the mechanism of substrate recognition of BoNT F by determining the crystal structures of its complex with two substrate-based inhibitors, VAMP 22-58/Gln58D-cysteine and 27-58/Gln58D cysteine. The inhibitors bind to BoNT F in the canonical direction (as seen for BoNTs A and E substrates) but are positioned specifically via three major exosites away from the active site. The cysteine sulfur of the inhibitors interacts with the zinc and exists as sulfinic acid in the inhibitor VAMP 27 58/Gln58D-cysteine. Arg133 and Arg171, which form part of two separate exosites, are crucial for substrate binding and catalysis. PMID- 19543289 TI - AID upmutants isolated using a high-throughput screen highlight the immunity/cancer balance limiting DNA deaminase activity. AB - DNA deaminases underpin pathways in antibody diversification (AID) and anti-viral immunity (APOBEC3s). Here we show how a high-throughput bacterial papillation assay can be used to screen for AID mutants with increased catalytic activity. The upmutations focus on a small number of residues, some highlighting regions implicated in AID's substrate interaction. Many of the upmutations bring the sequence of AID closer to that of APOBEC3s. AID upmutants can yield increased antibody diversification, raising the possibility that modification of AID's specific activity might be used to regulate antibody diversification in vivo. However, upmutation of AID also led to an increased frequency of chromosomal translocations, suggesting that AID's specific activity may have been limited by the risk of genomic instability. PMID- 19543290 TI - Control of alternative splicing through siRNA-mediated transcriptional gene silencing. AB - When targeting promoter regions, small interfering RNAs (siRNAs) trigger a previously proposed pathway known as transcriptional gene silencing by promoting heterochromatin formation. Here we show that siRNAs targeting intronic or exonic sequences close to an alternative exon regulate the splicing of that exon. The effect occurred in hepatoma and HeLa cells with siRNA antisense strands designed to enter the silencing pathway, suggesting hybridization with nascent pre-mRNA. Unexpectedly, in HeLa cells the sense strands were also effective, suggesting that an endogenous antisense transcript, detectable in HeLa but not in hepatoma cells, acts as a target. The effect depends on Argonaute-1 and is counterbalanced by factors favoring chromatin opening or transcriptional elongation. The increase in heterochromatin marks (dimethylation at Lys9 and trimethylation at Lys27 of histone H3) at the target site, the need for the heterochromatin-associated protein HP1alpha and the reduction in RNA polymerase II processivity suggest a mechanism involving the kinetic coupling of transcription and alternative splicing. PMID- 19543293 TI - Pediatrics: Intensive insulin therapy in critically ill children. PMID- 19543292 TI - Reproductive endocrinology: Male hormonal contraceptive passes efficacy test in China. PMID- 19543291 TI - Structure of a lamprey variable lymphocyte receptor in complex with a protein antigen. AB - Variable lymphocyte receptors (VLRs) are leucine-rich repeat proteins that mediate adaptive immunity in jawless vertebrates. VLRs are fundamentally different from the antibodies of jawed vertebrates, which consist of immunoglobulin (Ig) domains. We determined the structure of an anti-hen egg white lysozyme (HEL) VLR, isolated by yeast display, bound to HEL. The VLR, whose affinity resembles that of IgM antibodies, uses nearly all its concave surface to bind the protein, in addition to a loop that penetrates into the enzyme active site. The VLR-HEL structure combined with sequence analysis revealed an almost perfect match between ligand-contacting positions and positions with highest sequence diversity. Thus, it is likely that we have defined the generalized antigen-binding site of VLRs. We further demonstrated that VLRs can be affinity matured by 13-fold to affinities as high as those of IgG antibodies, making VLRs potential alternatives to antibodies for biotechnology applications. PMID- 19543294 TI - Adrenal function: An overlooked cause of glucocorticoid deficiency? PMID- 19543295 TI - Pancreas: C-peptide reveals possible beta-cell reactivation in pregnancy. PMID- 19543296 TI - Diabetes: Aspirin and prevention of diabetes still a topic of debate. PMID- 19543297 TI - Marine-derived oligosaccharide sulfate (JG3) suppresses heparanase-driven cell adhesion events in heparanase over-expressing CHO-K1 cells. AB - AIM: To elucidate the detailed mechanisms underlying the appreciable effects of JG3, a novel marine-derived oligosaccharide, on cell migration using a Chinese hamster ovary (CHO) cell line stably over-expressing heparanase. METHODS: A retrovirus infection system was used to establish a CHO-K1 cell line stably transfected with heparanase. Immunocytochemistry was used to assess cell morphology. Flow cytometry was selected to analyze the activation of beta1 integrin, and Western blotting was used to analyze the downstream effects on the cell adhesion pathway. An affinity precipitation assay was used to determine activation of the small GTPases, Rac1, and RhoA. RESULTS: JG3 abolished heparanase-driven formation of focal adhesions and cell spreading. Although JG3 failed to block the heparanase-triggered activation of beta1-integrin or the phosphorylation of Src, the oligosaccharide caused a significant dephosphorylation of FAK and subsequent inactivation of Erk. Furthermore, JG3 was found to arrest the activation of Rac1. CONCLUSION: All these findings help form an alternative view to understand the mechanisms underlying the inhibitory effects of JG3 on cell motility.Acta Pharmacologica Sinica (2009) 30: 1033-1038; doi: 10.1038/aps.2009.97; published online 22 June 2009. PMID- 19543298 TI - The expression of efflux and uptake transporters are regulated by statins in Caco 2 and HepG2 cells. AB - AIM: Statin disposition and response are greatly determined by the activities of drug metabolizing enzymes and efflux/ uptake transporters. There is little information on the regulation of these proteins in human cells after statin therapy. In this study, the effects of atorvastatin and simvastatin on mRNA expression of efflux (ABCB1, ABCG2 and ABCC2) and uptake (SLCO1B1, SLCO2B1 and SLC22A1) drug transporters in Caco-2 and HepG2 cells were investigated. METHODS: Quantitative real-time PCR was used to measure mRNA levels after exposure of HepG2 and Caco-2 cells to statins. RESULTS: Differences in mRNA basal levels of the transporters were as follows: ABCC2>ABCG2>ABCB1>SLCO1B1>>>SLC22A1>SLC O2B1 for HepG2 cells, and SLCO2B1>>ABCC2>ABCB1>ABCG2>>>SLC22A1 for Caco-2 cells. While for HepG2 cells, ABCC2, ABCG2 and SLCO2B1 mRNA levels were significantly up regulated at 1, 10 and 20 micromol/L after 12 or 24 h treatment, in Caco-2 cells, only the efflux transporter ABCB1 was significantly down-regulated by two-fold following a 12 h treatment with atorvastatin. Interestingly, whereas treatment with simvastatin had no effect on mRNA levels of the transporters in HepG2 cells, in Caco-2 cells the statin significantly down-regulated ABCB1, ABCC2, SLC22A1, and SLCO2B1 mRNA levels after 12 or 24 h treatment. CONCLUSION: These findings reveal that statins exhibits differential effects on mRNA expression of drug transporters, and this effect depends on the cell type. Furthermore, alterations in the expression levels of drug transporters in the liver and/or intestine may contribute to the variability in oral disposition of statins.Acta Pharmacologica Sinica (2009) 30: 956-964; doi: 10.1038/aps.2009.85; published online 22 June 2009. PMID- 19543299 TI - Pharmacokinetic studies of meloxicam following oral and transdermal administration in Beagle dogs. AB - AIM: The potential for topical delivery of meloxicam was investigated by examining its pharmacokinetic profiles in plasma and synovial fluid following oral and transdermal administration in Beagle dogs. METHODS: The experiment was a two-period, crossover design using 6 Beagle dogs. Meloxicam tablets were administered orally at a dose of 0.31 mg/kg, and meloxicam gel was administered transdermally at a dose of 1.25 mg/kg. Drug concentrations in plasma and synovial fluid were determined by liquid chromatography-tandem mass spectrometry (LC/MS/MS). The pharmacokinetic parameters were calculated using the Topfit 2.0 program. RESULTS: The pharmacokinetic results showed that AUC(0-t) (23.9+/-8.26 microg.h.mL(-1)) in plasma after oral administration was significantly higher than after transdermal delivery (1.00+/-0.43 microg.h.mL(-1)). In contrast, the ratio of the average concentration in synovial fluid to that in plasma following transdermal administration was higher than that for an oral delivery. The synovial fluid concentration in the treated leg was much higher than that in the untreated leg, whereas the synovial fluid concentration in the untreated leg was similar to the plasma concentration. CONCLUSION: The high concentration ratio of synovial fluid to plasma indicates direct penetration of meloxicam following topical administration to the target tissue. This finding is further supported by the differences observed in meloxicam concentrations in synovial fluid in the treated and untreated joints at the same time point. Our results suggest that transdermal delivery of meloxicam is a promising method for decreasing its adverse systemic effects.Acta Pharmacologica Sinica (2009) 30: 1060-1064; doi: 10.1038/aps.2009.73; published online 8 June 2009. PMID- 19543300 TI - MS80, a novel sulfated oligosaccharide, inhibits pulmonary fibrosis by targeting TGF-beta1 both in vitro and in vivo. AB - AIM: The pro-fibrogenic cytokine transforming growth factor-beta 1 (TGF-beta1) has attracted much attention for its potential role in the etiology of idiopathic pulmonary fibrosis (IPF). Here, we demonstrate that MS80, a novel sulfated oligosaccharide extracted from seaweed, can bind TGF-beta1. The aim of the present study was to determine whether MS80 is capable of combating TGF-beta1 mediated pulmonary fibrotic events both in vitro and in vivo, and to investigate the possible underlying mechanisms. METHODS: Surface plasmon resonance was used to uncover the binding profiles between the compound and TGF-beta. MTT assay, flow cytometry, Western blot analysis, BCA protein assay and SDS-PAGE gelatin zymography were used to probe the antifibrotic mechanisms of MS80. The in vivo fibrotic efficacy was evaluated in a bleomycin instillation-induced rat model. RESULTS: We report that MS80, a new kind of sulfated oligosaccharide extracted from seaweed, inhibits TGF-beta1-induced pulmonary fibrosis in vitro and bleomycin-induced pulmonary fibrosis in vivo. Our results indicated that MS80 competitively inhibited heparin/HS-TGF-beta1 interaction through its high binding affinity for TGF-beta1. Moreover, MS80 arrested TGF-beta1-induced human embryo pulmonary fibroblast (HEPF) cell proliferation, collagen deposition and matrix metalloproteinase (MMP) activity. Intriguingly, MS80 deactivated both the ERK and p38 signaling pathways. MS80 was also a potent suppressor of bleomycin-induced rat pulmonary fibrosis in vivo, as evidenced by improved pathological settings and decreased lung collagen contents. CONCLUSION: MS80 in particular, and perhaps oligosaccharide in general, offer better pharmacological profiles with appreciably few side effects and represent a promising class of drug candidates for IPF therapy.Acta Pharmacologica Sinica (2009) 30: 973-979; doi: 10.1038/aps.2009.86; published online 22 June 2009. PMID- 19543301 TI - Chronic intermittent hypobaric hypoxia protects the heart against ischemia/reperfusion injury through upregulation of antioxidant enzymes in adult guinea pigs. AB - AIM: To investigate the protection and the anti-oxidative mechanism afforded by chronic intermittent hypobaric hypoxia (CIHH) against ischemia/reperfusion (I/R) injury in guinea pig hearts. METHODS: Adult male guinea pigs were exposed to CIHH by mimicking a 5000 m high altitude (p(B)=404 mmHg, p(O2)=84 mmHg) in a hypobaric chamber for 6 h/day for 28 days. Langendorff-perfused isolated guinea pig hearts were used to measure variables of left ventricular function during baseline perfusion, ischemia and the reperfusion period. The activity and protein expression of antioxidant enzymes in the left myocardium were evaluated using biochemical methods and Western blotting, respectively. Intracellular reactive oxygen species (ROS) were assessed using ROS-sensitive fluorescence. RESULTS: After 30 min of global no-flow ischemia followed by 60 min of reperfusion, myocardial function had better recovery rates in CIHH guinea pig hearts than in control hearts. The activity and protein expression of superoxide dismutase (SOD) and catalase (CAT) were significantly increased in the myocardium of CIHH guinea pigs. Pretreatment of control hearts with an antioxidant mixture containing SOD and CAT exerted cardioprotective effects similar to CIHH. The irreversible CAT inhibitor aminotriazole (ATZ) abolished the cardioprotection of CIHH. Cardiac contractile dysfunction and oxidative stress induced by exogenous hydrogen peroxide (H(2)O(2)) were attenuated by CIHH and CAT. CONCLUSIONS: These data suggest that CIHH protects the heart against I/R injury through upregulation of antioxidant enzymes in guinea pig.Acta Pharmacologica Sinica (2009) 30: 947-955; doi: 10.1038/aps.2009.57; published online 22 June 2009. PMID- 19543302 TI - BMP6 reverses TGF-beta1-induced changes in HK-2 cells: implications for the treatment of renal fibrosis. AB - AIM: The aim of the study was to investigate the potential role of BMP6 in TGF beta1-mediated changes in HK-2 cells. METHODS: BMP6 was purified via heparin affinity and reverse phase liquid chromatography. The purity, specificity, and bioactivity of BMP6 were determined by SDS-PAGE, Western blot assays, and the induction of alkaline phosphatase (ALP) activity, respectively. Cell proliferation, morphology, and expression levels of alpha-SMA and E-cadherin were assessed by cell viability, microscopy, and Western blot assays, respectively. In addition, cell adhesion abilities were determined by counting the number of attached cells. The expression of fibronectin, collagen IV, matrix metalloproteinases 2 (MMP-2), and tissue inhibitors of matrix metalloproteinases 2 (TIMP-2) were analyzed using RT-PCR. MMP-2 activity was analyzed by zymography, whereas the activation of the MAPKs and Smad signaling were analyzed using Western blot assays and a reporter gene assay, respectively. RESULTS: Our results indicated that recombinant BMP6 induced ALP activity in a dose-dependent and time course-dependent manner. Treatment with TGF-beta1 reduced both the cell proliferation and the expression of E-cadherin, induced a morphological transformation, decreased the expression and activity of MMP-2, and increased the expression levels of alpha-SMA, fibronectin, and TIMP-2 in HK-2 cells. All of these effects were inhibited when cells were treated with TGF-beta1 in combination with rhBMP6, whereas rhBMP6 alone demonstrated no such effect. Treatment with TGF-beta1, rhBMP6, or a combination of both had no effect on the expression of collagen IV. In addition, the administration of rhBMP6 prevented the enhanced adhesion behavior triggered by TGF-beta1. Furthermore, the addition of rhBMP6 abrogated the JNK and Smad2/3 signaling that was activated by TGF beta1. CONCLUSION: BMP6 ameliorated the TGF-beta1-induced changes in HK-2 cells. The suppression of TGF-beta1-mediated JNK and Smad2/3 signaling activation were implicated in these effects.Acta Pharmacologica Sinica (2009) 30: 994-1000; doi: 10.1038/aps.2009.56; published online 22 June 2009. PMID- 19543304 TI - Racial differences and cardiovascular response to psychological stress. PMID- 19543306 TI - Value for money. AB - 2009 is turning out to be an interesting year for research funding in the UK. Everyone wants the best science to be funded, but it's not clear that the proposed policy changes will achieve this. PMID- 19543305 TI - Genetic and pharmacogenetic associations between NOS3 polymorphisms, blood pressure, and cardiovascular events in hypertension. PMID- 19543308 TI - Metamaterials: Towards invisibility in the visible. PMID- 19543309 TI - Organic conductors: Polymers as one-dimensional metals. PMID- 19543310 TI - DNA nanotechnology: Hot and sticky or cold and aloof. PMID- 19543311 TI - Nanocontacts: The importance of being entangled. PMID- 19543313 TI - Alkali metal crystalline polymer electrolytes. AB - Polymer electrolytes have been studied extensively because uniquely they combine ionic conductivity with solid yet flexible mechanical properties, rendering them important for all-solid-state devices including batteries, electrochromic displays and smart windows. For some 30 years, ionic conductivity in polymers was considered to occur only in the amorphous state above Tg. Crystalline polymers were believed to be insulators. This changed with the discovery of Li(+) conductivity in crystalline poly(ethylene oxide)(6):LiAsF(6). However, new crystalline polymer electrolytes have proved elusive, questioning whether the 6:1 complex has particular structural features making it a unique exception to the rule that only amorphous polymers conduct. Here, we demonstrate that ionic conductivity in crystalline polymers is not unique to the 6:1 complex by reporting several new crystalline polymer electrolytes containing different alkali metal salts (Na(+), K(+) and Rb(+)), including the best conductor poly(ethylene oxide)(8):NaAsF(6) discovered so far, with a conductivity 1.5 orders of magnitude higher than poly(ethylene oxide)(6):LiAsF(6). These are the first crystalline polymer electrolytes with a different composition and structures to that of the 6:1 Li(+) complex. PMID- 19543314 TI - Rational design and application of responsive alpha-helical peptide hydrogels. AB - Biocompatible hydrogels have a wide variety of potential applications in biotechnology and medicine, such as the controlled delivery and release of cells, cosmetics and drugs, and as supports for cell growth and tissue engineering. Rational peptide design and engineering are emerging as promising new routes to such functional biomaterials. Here, we present the first examples of rationally designed and fully characterized self-assembling hydrogels based on standard linear peptides with purely alpha-helical structures, which we call hydrogelating self-assembling fibres (hSAFs). These form spanning networks of alpha-helical fibrils that interact to give self-supporting physical hydrogels of >99% water content. The peptide sequences can be engineered to alter the underlying mechanism of gelation and, consequently, the hydrogel properties. Interestingly, for example, those with hydrogen-bonded networks of fibrils melt on heating, whereas those formed through hydrophobic fibril-fibril interactions strengthen when warmed. The hSAFs are dual-peptide systems that gel only on mixing, which gives tight control over assembly. These properties raise possibilities for using the hSAFs as substrates in cell culture. We have tested this in comparison with the widely used Matrigel substrate, and demonstrate that, like Matrigel, hSAFs support both growth and differentiation of rat adrenal pheochromocytoma cells for sustained periods in culture. PMID- 19543315 TI - GATA4 is a regulator of astrocyte cell proliferation and apoptosis in the human and murine central nervous system. AB - The GATA transcription factors consist of six family members, which bind to the consensus DNA-binding element, W-GATA-R, and are poorly characterized in the central nervous system (CNS). Using retroviral gene trapping on transgenic mouse glioma models, we identified GATA6 to be a novel tumor suppressor gene in glioblastoma multiforme. We now show GATA4, a family member of GATA6, to be expressed in the neurons and glia of normal murine and human embryonic and adult CNS. Silencing GATA4 in normal astrocytes did not alter their growth properties. In contrast, knockdown of Gata4 in p53 null non-transformed murine astrocytes induced transformation, with increased proliferation and resistance to chemotherapy or radiation-induced apoptosis. Furthermore, GATA4 expression was lost in a panel of human malignant astrocytoma cell lines. GATA4 overexpression in normal human and murine astrocytes resulted in a cell cycle block in G1 phase, with increased apoptosis. Mechanistically, GATA4 was a transcriptional inducer of the cyclin-dependent kinase inhibitor, p15INK4B, leading to the attenuation of cyclin D1. GATA4 expression was also induced by transforming growth factor-beta, leading to the inhibition of astrocyte proliferation. Collectively, we show that GATA4 is expressed in the embryonic and adult CNS and acts as a negative regulator of astrocyte proliferation and growth. PMID- 19543316 TI - SOCS-mediated downregulation of mutant Jak2 (V617F, T875N and K539L) counteracts cytokine-independent signaling. AB - Recently, mutations in the gene of Janus kinase 2 (Jak2) were discovered in patients suffering from chronic myeloproliferative disorders (MPD) and leukemia. As suppressors of cytokine signaling (SOCS) proteins are potent feedback inhibitors of Jak-mediated signaling, we investigated their role in signal transduction through constitutively active Jak2 mutants. We selected two mutants, Jak2-V617F and Jak2-K539L, found in patients with MPDs and Jak2-T875N identified in acute megakaryoblastic leukemia. We found SOCS family members to be induced through Jak2-V617F in human leukemia cell lines expressing the mutant allele and in stable HEK transfectants inducibly expressing constitutively active Jak2 mutants. SOCS proteins were recruited to the membrane and bound to the constitutively active Jaks. In contrast to wild-type Jak2, the mutant proteins were constitutively ubiquitinated and degraded through the proteasome. Taken together, we show a SOCS-mediated downregulation of the constitutively active, disease-associated mutant Jak2 proteins. Furthermore, a threshold level of mutant Jak expression has to be overcome to allow full cytokine-independent constitutive activation of signaling proteins, which may explain progression to homozygocity in MPDs as well as gene amplification in severe phenotypes and leukemia. PMID- 19543318 TI - Alterations of ubiquitin ligases in human cancer and their association with the natural history of the tumor. AB - Protein ubiquitination is critical for many cellular processes, through its ability to regulate protein degradation and various signaling mechanisms. In the ubiquitin (Ub) system, substrate specificity is achieved through the E3 family of Ub ligases. Because alterations of the ubiquitination machinery have been reported in human cancers, the selective interference with Ub ligases might represent a powerful therapeutic tool. Here, we report the first wide survey of misregulation of Ub ligases in cancer. We analysed 82 Ub ligases in nine types of cancer by in situ hybridization on tissue microarrays. We found 27 instances in which an Ub ligase was altered in a given type of tumor, when compared with normal tissues: 21 cases of overexpression and 6 cases of underexpression. We further analysed selected Ub ligases in large cohorts of breast and non-small cell lung carcinomas. In five, of six, of these extended analyses (HUWE1, CCNB1IP1, SIAH1 and SIAH2 in breast cancer and CCNB1IP1 in lung cancer), we found that the levels of Ub ligases correlated significantly with relevant prognostic factors, and with clinical outcome. Our findings show that the alteration of Ub ligases is a frequent event in cancer and identify candidate targets for molecular therapies. PMID- 19543317 TI - BMI1 cooperates with H-RAS to induce an aggressive breast cancer phenotype with brain metastases. AB - B-lymphoma Moloney murine leukaemia virus insertion region-1 (BMI1) is a member of the polycomb group of transcription repressors, which functions in stem cell maintenance and oncogenesis through the inhibition of the INK4A/ARF tumour suppressor locus. Overexpression of BMI1 is associated with poor prognosis in several human cancers, including breast cancer. We have previously shown that BMI1 collaborates with H-RAS to induce transformation of MCF10A human mammary epithelial cells through dysregulation of multiple growth pathways independent of the INK4A/ARF locus. In this study, we show that BMI1 collaborates with H-RAS to promote increased proliferation, invasion and resistance to apoptosis in vitro, and an increased rate of spontaneous metastases from mammary fat pad xenografts including novel metastases to the brain. Furthermore, in collaboration with H RAS, BMI1 induced fulminant metastatic disease in the lung using a tail vein model of haematogenous spread through accelerated cellular proliferation and inhibition of apoptosis. Finally, we show that knockdown of BMI1 in several established breast cancer cell lines leads to decreased oncogenic behaviour in vitro and in vivo. In summary, BMI1 collaborates with H-RAS to induce an aggressive and metastatic phenotype with the unusual occurrence of brain metastasis, making it an important target for diagnosis and treatment of aggressive breast cancer. PMID- 19543319 TI - Cyclic GMP-dependent protein kinase II inhibits cell proliferation, Sox9 expression and Akt phosphorylation in human glioma cell lines. AB - Earlier we used a glioma model to identify loci in the mouse genome, which were repeatedly targeted by platelet-derived growth factor (PDGF)-containing Moloney murine leukemia viruses. The gene Prkg2, encoding cyclic guanosine monophosphate (cGMP)-dependent protein kinase II, cGKII, was tagged by retroviral insertions in two brain tumors. The insertions were both situated upstream of the kinase domain and suggested creating a truncated form of the cGKII protein. We transfected different human glioma cell lines with Prkg2 and found an overall reduction in colony formation and cell proliferation compared with controls transfected with truncated Prkg2 (lacking the kinase domain) or empty vector. All glioma cells transfected with the cGKII phosphorylate vasodilator-stimulated phosphoprotein, VASP, after cGMP analog treatment. Glioma cell lines positive for the Sox9 transcription factor showed reduced Sox9 expression when Prkg2 was stably transfected. When cGKII was activated by cGMP analog treatment, Sox9 was phosphorylated, Sox9 protein expression was suppressed and the glioma cell lines displayed loss of cell adhesion, inhibition of Akt phosphorylation and G1 arrest. Sox9 repression by siRNA was similarly shown to reduce glioma cell proliferation. Expression analysis of stem and glial lineage cell markers also suggests that cGKII induces differentiation of glioma cell lines. These findings describe an anti-proliferative role of cGKII in human glioma biology and would further explain the retroviral tagging of the cGKII gene during brain tumor formation in PDGF-induced tumors. PMID- 19543321 TI - XEDAR as a putative colorectal tumor suppressor that mediates p53-regulated anoikis pathway. AB - Colorectal cancers with mutations in the p53 gene have an invasive property, but its underlying mechanism is not fully understood. Through the screening of two data sets of the genome-wide expression profile, one for p53-introduced cells and the other for the numbers of cancer tissues, we report here X-linked ectodermal dysplasia receptor (XEDAR), a member of the TNFR superfamily, as a novel p53 target that has a crucial role in colorectal carcinogenesis. p53 upregulated XEDAR expression through two p53-binding sites within intron 1 of the XEDAR gene. We also found a significant correlation between decreased XEDAR expressions and p53 gene mutations in breast and lung cancer cell lines (P=0.0043 and P=0.0122, respectively). Furthermore, promoter hypermethylation of the XEDAR gene was detected in 20 of 20 colorectal cancer cell lines (100%) and in 6 of 12 colorectal cancer tissues (50%), respectively. Thus, the XEDAR expression was suppressed to <25% of surrounding normal tissues in 12 of 18 colorectal cancer tissues (66.7%) due to either its epigenetic alterations and/or p53 mutations. We also found that XEDAR interacted with and subsequently caused the accumulation of FAS protein, another member of p53-inducible TNFR. Moreover, XEDAR negatively regulated FAK, a central component of focal adhesion. As a result, inactivation of XEDAR resulted in the enhancement of cell adhesion and spreading, as well as resistance to p53-induced apoptosis. Taken together, our findings showed that XEDAR is a putative tumor suppressor that could prevent malignant transformation and tumor progression by regulating apoptosis and anoikis. PMID- 19543320 TI - Proteinase-activated receptor 2 expression in breast cancer and its role in breast cancer cell migration. AB - Proteinase-activated receptor 2 (PAR2) is a G protein-coupled receptor that is activated by trypsin-like proteinases. PAR2 is detected in breast tumor specimens; however, it is not clear how PAR2 level in breast cancer cell/tissues compares with normal cell/tissues. Here, we show the elevation of PAR2 protein level in 76 of 105 breast tumor specimens but only 5 of 24 normal breast tissues. PAR2 level is also higher in breast cancer cell lines than that in normal breast cells and non-cancerous breast cell lines. To determine the role of PAR2 in breast carcinogenesis, we examined the effect of PAR2 agonists on cell proliferation and migration. Our studies show that PAR2 agonists (PAR2-activating peptide and trypsin) are neither potent growth enhancers nor chemoattractants to breast cancer cells. Instead, PAR2 agonists induce significant chemokinesis. PAR2 mediated chemokinesis is G(alphai)-dependent, and inhibiting Src kinase activity or silencing c-Src expression blocks PAR2-mediated chemokinesis. These results suggest that c-Src works downstream of G(alphai) to mediate this PAR2 agonist induced event. To characterize c-Src effector, we reveal that PAR2 agonists activate JNKs in a Src-dependent manner and that JNK activity is essential for PAR2-mediated chemokinesis. Moreover, PAR2 agonist stimulation leads to paxillin Ser(178) phosphorylation and paxillin(S178A) mutant inhibits PAR2-mediated chemokinesis. In conclusion, our studies show that PAR2 agonists facilitate breast cancer cell chemokinesis through the G(alphai)-c-Src-JNK-paxillin signaling pathway. PMID- 19543323 TI - Malonyl-CoA decarboxylase inhibition is selectively cytotoxic to human breast cancer cells. AB - Fatty acid synthase (FAS) inhibition initiates selective apoptosis of cancer cells both in vivo and in vitro, which may involve malonyl-CoA metabolism. These findings have led to the exploration of malonyl-CoA decarboxylase (MCD) as a potential novel target for cancer treatment. MCD regulates the levels of cellular malonyl-CoA through the decarboxylation of malonyl-CoA to acetyl-CoA. Malonyl-CoA is both a substrate for FAS and an inhibitor of fatty acid oxidation acting as a metabolic switch between anabolic fatty acid synthesis and catabolic fatty acid oxidation. We now report that the treatment of human breast cancer (MCF7) cells with MCD small interference RNA (siRNA) reduces MCD expression and activity, reduces adenosine triphosphate levels, and is cytotoxic to MCF7 cells, but not to human fibroblasts. In addition, we synthesized a small-molecule inhibitor of MCD, 5-{(Morpholine-4-carbonyl)-[4-(2,2,2-trifluoro-1-hydroxy-1-trifluoromethyl-ethyl) phenyl]-amino}-pentanoic acid methyl ester (MPA). Similar to MCD siRNA, MPA inhibits MCD activity in MCF7 cells, increases cellular malonyl-CoA levels and is cytotoxic to a number of human breast cancer cell lines in vitro. Taken together, these data indicate that MCD-induced cytotoxicity is likely mediated through malonyl-CoA metabolism. These findings support the hypothesis that MCD is a potential therapeutic target for cancer therapy. PMID- 19543322 TI - A combinatorial mechanism for determining the specificity of E2F activation and repression. AB - Various studies have detailed the role of E2F proteins in both transcription activation and repression. Further study has shown that distinct promoter elements, but comprising the same E2F-recognition motif, confer positive or negative E2F control and that this reflects binding of either activator or repressor E2F proteins, respectively. We now show that the specificity of binding of an activator or repressor E2F protein is determined by adjacent sequences that bind a cooperating transcription factor. We propose that the functional E2F element is a module comprising not only the E2F-binding site but also the adjacent site for the cooperating transcription factor. PMID- 19543325 TI - Reduced-intensity conditioning using fludarabine with either antithymocyte globulin and BU, or low-dose TBI allowing allogeneic hematopoietic SCT. AB - In a single-center study, we analyzed the outcomes of 66 patients with advanced hematological malignancies receiving two reduced-intensity conditioning regimens for allogeneic transplantation: fludarabine and low-dose TBI (flu/TBI, n=25), or fludarabine, antithymocyte globulin and BU (flu/ATG/BU, n=41). The selection criteria were based on the hypothesis that flu/TBI patients were expected to achieve autologous recovery in the event of non-engraftment. Sixty-three patients (95%) engrafted. Regimen-related mortality at day 100 and 1 year was 6 and 15%, respectively. With median follow-up of 50.4 months, survival did not differ by regimen. Multivariate analysis confirmed that the type of regimen did not affect relapse. In patients achieving full donor chimerism by day 30, those conditioned with flu/TBI showed greater overall survival (P=0.02). Engraftment failure occurred in two patients (3%), both of whom received flu/TBI. We conclude that conditioning with flu/TBI or flu/ATG/BU yields comparable survival and remission outcomes. By contrast to our hypothesis, patients receiving flu/TBI who subsequently failed engraftment did not achieve autologous recovery. Yet, rapid attainment of full donor chimerism after flu/TBI is associated with greater survival than after flu/ATG/BU. Further, larger prospective randomized studies are required to define the advantage of one regimen over the other. PMID- 19543324 TI - Significance of RRM1 and ERCC1 expression in resectable pancreatic adenocarcinoma. AB - The identification of molecular markers, useful for therapeutic decisions in pancreatic cancer patients, is crucial for advances in disease management. Gemcitabine, although a cornerstone of current therapy, has limited efficacy. RRM1 is a key molecule for gemcitabine efficacy and is also involved in tumor progression. We determined in situ RRM1 and excision repair cross complementation group 1 (ERCC1) protein levels in 68 pancreatic cancer patients. All had R0 resections without preoperative therapy. Protein levels were determined by automated quantitative analysis (AQUA), a fluorescence-based immunohistochemical method. The relationship between protein expressions and clinical outcomes, including response to gemcitabine at the time of disease recurrence, was determined. Patients with high RRM1 showed significantly better overall survival than patients with low expression (P=0.0196). There was a trend toward better overall survival for patient with high ERCC1 (P=0.0552). When both markers were considered together, patients with both high RRM1 and ERCC1 faired the best in terms of overall and disease-free survival (P=0.0066, P=0.0127). In addition, treatment benefit from gemcitabine in patients with disease recurrence was observed only in patients with low RRM1. The combination of RRM1 and ERCC1 expression is prognostic in pancreatic cancer patients after a complete resection. On disease recurrence, only patients with low RRM1 derive benefit from gemcitabine. PMID- 19543326 TI - Unrelated umbilical cord blood transplantation in children with immune deficiency: results of a multicenter study. AB - In the absence of a related donor, unrelated cord blood transplant (CBT) may be a potential option for patients with a primary immune deficiency (PID). Most published experience consists of single-center data using multiple preparative regimens and GVHD prophylaxis. We report the results of a multicenter prospective trial of unrelated CBT for PID. A total of 24 children with PID, with a median age of 1 year (range: 0.23-7.81 years) and a median weight of 10.5 kg (range: 4 24.4 kg) received unrelated CBT between 1999 and 2003. All patients received a fully ablative conditioning regimen with identical GVHD prophylaxis and supportive care. Most patients (79%) received a 1 or 2 HLA Ag-mismatched cord unit with a median nucleated cell infused of 9.3 x 10(7)/kg (range: 1.0-31.2) and a median CD34 of 2.7 x 10(5)/kg 2.9 (range: 0.6-84.5). The cumulative incidence of neutrophil engraftment by day 42 was 58% (95% CI: 38-79%) at a median of 19 days. Cumulative incidence estimates of grade III-IV acute GVHD at day 100 and chronic GVHD at 1 year were 29% (95% CI: 10-48%) and 24% (95% CI: 3-44%), respectively. The probability of survival at 180 days and 1 year was 66.7% (95% CI: 44.3-81.7%) and 62.5% (95% CI: 40.3-78.4%), respectively. Unrelated CBT should be considered in children with PID. PMID- 19543328 TI - Isolated testicular relapse after allo-SCT in boys with ALL: outcome without second transplant. PMID- 19543327 TI - 5-Azacitidine for myelodysplasia before allogeneic hematopoietic cell transplantation. AB - Relapse remains a leading cause for treatment failure after hematopoietic cell transplantation (HCT) in patients with intermediate- or high-risk myelodysplastic syndrome (MDS). To discern the impact of 5-azacitine treatment pretransplant on the risk for relapse after HCT, we analyzed the post transplant outcomes of all 54 consecutive patients with MDS or chronic myelomonocytic leukemia who received HCT from HLA-compatible donors according to pretransplant 5-azacitidine exposure. Thirty patients received a median of four (1-7) cycles of 5-azacitidine, and 24 patients did not receive 5-azacitidine before HCT. The 1-year estimates of overall survival, relapse-free survival and cumulative incidence of relapse were 47, 41 and 20%, for 5-azacitidine patients and 60, 51 and 32%, respectively, for non-5-azacytidine patients. These observations suggest that outcomes are similar in both groups with a trend toward decreased early relapse in patients receiving 5-azacitidine. 5-Azacitidine may be of value in stabilizing the disease, thereby allowing time for patients to reach transplant and does not appear to affect transplant outcomes. PMID- 19543329 TI - T-cell clones persisting in the circulation after autologous hematopoietic SCT are undetectable in the peripheral CD34+ selected graft. AB - In addition to its established hematological indications, autologous hematopoietic SCT (HSCT) can ameliorate the course of severe autoimmune disorders through a reconditioning of the immune system. We have shown earlier that HSCT determines extensive renewal of the TCR repertoire in multiple sclerosis patients. However, the observed persistence post-therapy of some pre-existing T cell clones suggested the potential for disease recapitulation. Here, we investigated whether TCRs that reappear after a myeloablative conditioning regimen and HSCT were reintroduced with the autologous, CD34-selected hematopoietic stem cell (HSC) graft. In all, we cloned and sequenced 2237 TCR clones from peripheral blood and HSC grafts from four patients who underwent autologous HSCT for severe multiple sclerosis. Surprisingly, in-frame TCR sequences were detectable in only one of four patient grafts and no TCR sequences were found to be shared between the graft and pre- or post-HSCT samples. These findings provide the first evidence from extensive sequencing analysis to suggest that T cells in autologous HSC grafts that have been mobilized with CY+G-CSF and CD34-selected have limited survival capacity and are therefore unlikely to be a major source of carryover of T-cell expansions potentially involved in autoimmune disease. PMID- 19543330 TI - Safety and efficacy assessment of plerixafor in patients with multiple myeloma proven or predicted to be poor mobilizers, including assessment of tumor cell mobilization. AB - This was an open-label, single-center, phase II study of 20 patients with multiple myeloma who were either proven poor mobilizers (n=10; group A) or predicted poor mobilizers (n=10; group B) and were planned for autologous hematopoietic SCT. The aim was to assess the safety and efficacy of plerixafor for stem cell mobilization and tumor cell contamination. The peripheral blood (PB) CD34+ cell count was generally very low pre- plerixafor and increased significantly post-plerixafor administration. Cumulative apheresis yields of > or =2 x 10(6) CD34+ cells/kg were observed in 7 of 10 patients (group A) and 8 of 10 patients (group B). Among the proven poor mobilizers, there was no evidence of tumor cell mobilization in the PB after G-CSF plus plerixafor treatment. Seventeen of 20 (85%) patients underwent transplantation. Neutrophil engraftment occurred at a median of 13 days for all patients. Platelet engraftment occurred at a median of 16 days and 19 days for all proven and predicted poor mobilizers, respectively. At 12 months, 12 of 17 patients had documented durable grafts, 3 of 17 patients died and 2 of 17 patients were lost to follow-up; but they had documented graft durability at the previous 3- and 6-month visit. The safety profile of plerixafor in all patients was consistent with previous reports. PMID- 19543332 TI - Non-turning-point monitoring improves narrow bandpass filters. AB - Historic practice has been to optically monitor narrow bandpass filters by the termination of each layer at a turning point. The problem is that turning points are the least sensitive points to the change of the optical signal with thickness and, thereby, those points are most prone to errors. It is shown that better performance in the production results can be achieved by designing and monitoring in order to terminate layers at non-turning points. A further advantage has been discovered wherein nonoptical monitoring of some layers is used to achieve even better stability in the production result. Simulation programs have been applied to such designs and demonstrate the advantages as compared to the historical approach. PMID- 19543331 TI - Reliability of a pretransplant i.v. BU test dose performed 2 weeks before myeloablative FluBu conditioning regimen. AB - A pretransplant test dose of i.v. BU was previously used in pediatric patients undergoing a reduced-intensity allogeneic hematopoietic SCT (HSCT). Here, we used a BU test dose in 23 adult patients who were not pancytopenic and underwent a myeloablative allogeneic HSCT prepared with fludarabine and i.v. BU (FluBU). Pharmacokinetics (PK) of BU were calculated after a test dose (0.8 mg/kg) was performed 2 weeks before transplant. Targeted BU area under the curve (AUC) range was 4800-5200 microM min. The mean BU dose calculated after the test dose was 3.5+/-0.5 mg/kg. To validate the test dose, PK studies were repeated in 17 patients after the first dose of BU during the conditioning regimen. An AUC below the therapeutic value of 4000 microM min was observed in 23% of the patients receiving a wt-based dose and in 0% of patients whose dose was calculated on the basis of the test dose (P=0.03). In patients who had a test dose, a significant correlation (P<0.0001) between the first and subsequent doses of BU during the conditioning regimen was observed. Our findings may allow more centers to pursue transplant strategies with targeted BU by overcoming the time limitation for PK studies during the conditioning regimen. PMID- 19543333 TI - Flexible meniscus/biconvex lens system with fluidic-controlled tunable-focus applications. AB - Controlled injection of fluid into a flexible lens chamber has previously produced tunable spherical lenses with spherical aberration problems. This paper presents a novel fluidic-controlled meniscus/biconvex lens system with relatively low distortion. The lens fabrication method is described and manufactured lenses are characterized for different processing parameters. Images captured through one of our lenses for a range of different pressures are presented. A change in focal length of five times (19.1-80.8 cm) is demonstrated. The focal length shows a quadratic relation to the applied pressure (0-5 kPa). For the simple prototype lens, distortion is less than 5% and relative illumination is above 0.74 by ZEMAX analysis. PMID- 19543334 TI - Expansion of field of view in digital in-line holography with a programmable point source. AB - We present a technique for programming the source of the spherical reference illumination in digital in-line holography using digital micromirror devices. The programmable point source is achieved by individually addressing the elements of a digital micromirror device to spatially control the illumination of the object located at some distance from the source of the spherical reference field. By moving the location of the "ON" element on the digital micromirror device, translation of both the source of the spherical reference beam and the captured holograms is achieved. Results obtained through numerical reconstruction of these translated holograms shows the possibility of expanding the field of view by about 263%. PMID- 19543335 TI - Correlation method for the measure of mask-induced line-edge roughness in extreme ultraviolet lithography. AB - As critical dimensions for leading-edge semiconductor devices shrink, the line edge roughness (LER) requirements are pushing well into the single digit nanometer regime. At these scales many new sources of LER must be considered. In the case of extreme ultraviolet (EUV) lithography, modeling has shown the lithographic mask to be a source of significant concern. Here we present a correlation-based methodology for experimentally measuring the magnitude of mask contributors to printed LER. The method is applied to recent printing results from a 0.3 numerical aperture EUV microfield exposure tool. The measurements demonstrate that such effects are indeed present and of significant magnitude. The method is also used to explore the effects of illumination coherence and defocus and has been used to verify model-based predictions of mask-induced LER. PMID- 19543336 TI - Phase reconstruction of digital holography with the peak of the two-dimensional Gabor wavelet transform. AB - We describe a numerical reconstruction technique for digital holography by means of the two-dimensional Gabor wavelet transform (2D-GWT). Applying the 2D-GWT to digital holography, the object wave can be reconstructed by calculating the wavelet coefficients of the hologram at the peak of the 2D-GWT automatically. At the same time the effect of the zero-order diffraction image and the twin image are eliminated without spatial filtering. Comparing the numerical reconstruction of a holographic image by the analysis of the one-dimensional Gabor wavelet transform (1D-GWT) with the 2D-GWT, we show that the 2D-GWT method is superior to the 1D-GWT method, especially when the fringes of the hologram are not just along the y direction. The theory and the results of a simulation and experiments are shown. PMID- 19543337 TI - Design and characterization of a nonuniform linear vertical-cavity surface emitting laser array with a Gaussian far-field distribution. AB - A 980 nm bottom-emitting vertical-cavity surface-emitting laser array with a nonuniform linear arrangement is reported to realize emission with a Gaussian far field distribution. This array is composed of five symmetrically arranged elements of 200 microm, 150 microm, and 100 microm diameter, with center spacing of 300 microm and 250 microm, respectively. An output power of 880 mW with a high power density of 1 kW/cm2 is obtained. The divergence angle is below 20 degrees in the range of operating current from 0 A to 6 A. The theoretical simulation of the near-field and the far-field distribution is in good agreement with the experimental result. The comparison between this nonuniform linear array, the single device, and the conventional two-dimensional array is carried out to demonstrate the good performance of the linear array. PMID- 19543338 TI - Retrievals of atmospheric CO2 from simulated space-borne measurements of backscattered near-infrared sunlight: accounting for aerosol effects. AB - Retrievals of atmospheric carbon dioxide (CO2) from space-borne measurements of backscattered near-infrared sunlight are hampered by aerosol and cirrus cloud scattering effects. We propose a retrieval approach that allows for the retrieval of a few effective aerosol parameters simultaneously with the CO2 total column by parameterizing particle amount, height distribution, and microphysical properties. Two implementations of the proposed method covering different spectral bands are tested for an ensemble of simulated nadir observations for aerosol (and cirrus) loaded scenes over low- and mid-latitudinal land surfaces. The residual aerosol-induced CO(2) errors are mostly below 1% up to aerosol optical thickness 0.5. The proposed methods also perform convincing for scenes where cirrus clouds of optical thickness 0.1 overlay the aerosol. PMID- 19543339 TI - Theoretical and experimental analysis of the gating performance of a photocathode gated image tube using a femtosecond laser. AB - We study theoretically and experimentally the gating performance of photocathode gated image tube. A cross-correlation method is proposed to analyze the rising and falling speed and width of the image tube gain. Femtosecond pulses generated by a fiber laser are used as the light source of ultrahigh temporal resolution and trapezoid electrical signals are applied to a photocathode electrode as gating pulses. By adjusting the time delay between the laser pulse and the electrical gating pulse, various acceleration procedures for the photoelectrons generated at the photocathode can be observed. The photoelectrons arriving at the multichannel plate (MCP) with different kinetic energies receive different gain according to Eberhardt's MCP gain model. The gain profile is obtained by measuring the output light power of the fluorescent screen at the output port of the tube. The theoretical analysis and experimental result show that the shape of the output gain curve of the image tube is deformed and the width is broadened in comparison with the symmetric electrical gating pulse. PMID- 19543340 TI - Four-dimensional measurement by a single-frame structured light method. AB - A four-dimensional [4D--three-dimensional (3D) shape varying in time] shape measurement system is described. A single 3D shape of an object is calculated from only one frame. The projected pattern is composed of sinusoidal intensity fringes and one color-encoded stripe, the analysis of which allows us to find the absolute coordinates of the measured object. During measurement, the position of the stripe changes due to the improvement of the quality of spatiotemporal unwrapping. The fringes deformed by the shape of the object are captured by a CCD camera and processed by an adaptive spatial carrier phase-shifting algorithm. The use of an algorithm based on fast Fourier transformation is proposed to approximate the local period of fringes. A new phase-unwrapping routine based on the spatiotemporal information is presented as well. All these features make the 3D shape measurement of an object in motion possible with the additional advantage of using a low-cost system. Experimental results of the developed method together with a preliminary assessment of measurement uncertainty are presented to show the validity of the method. PMID- 19543341 TI - Development of an ambient pressure laser-induced fluorescence instrument for nitrogen dioxide. AB - Concerns about the health effects of nitrogen dioxide (NO2) and its role in forming deleterious atmospheric species have made it desirable to have low-cost, sensitive ambient measurements of NO2. We have developed a continuous-wave laser diode laser-induced fluorescence (LIF) system for NO2 that operates at ambient pressure, thereby eliminating the need for an expensive pumping system. The current prototype system has achieved sensitivity several orders of magnitude beyond previous efforts at ambient pressure (limit of detection of 2 ppb, 60 s averaging time). Ambient measurements of NO2 were made in Portland, Oregon using both the standard NO2 chemiluminescence method and the LIF instrument and showed good agreement (r2 = 0.92). PMID- 19543342 TI - Cerenkov light spectrum in an optical fiber exposed to a photon or electron radiation therapy beam. AB - A Cerenkov signal is generated when energetic charged particles enter the core of an optical fiber. The Cerenkov intensity can be large enough to interfere with signals transmitted through the fiber. We determine the spectrum of the Cerenkov background signal generated in a poly(methyl methacrylate) optical fiber exposed to photon and electron therapeutic beams from a linear accelerator. This spectral measurement is relevant to discrimination of the signal from the background, as in scintillation dosimetry using optical fiber readouts. We find that the spectrum is approximated by the theoretical curve after correction for the wavelength dependent attenuation of the fiber. The spectrum does not depend significantly on the angle between the radiation beam and the axis of the fiber optic but is dependent on the depth in water at which the fiber is exposed to the beam. PMID- 19543343 TI - Demonstration of an infrared microcamera inspired by Xenos peckii vision. AB - We present an original and compact optical system inspired by the unusual eyes of a Strepsipteran insect called Xenos peckii. It is designed for a field of view of 30 degrees and is composed of multiple telescopes. An array of prisms of various angles is placed in front of these telescopes in order to set a different field of view for each channel. This type of camera operates in the [3-5 microm] spectral bandwidth and is entirely integrated in a Dewar in order to maximize its compactness. Experimental images are presented to validate this design. PMID- 19543344 TI - Low cost, high performance, self-aligning miniature optical systems. AB - The most expensive aspects in producing high quality miniature optical systems are the component costs and long assembly process. A new approach for fabricating these systems that reduces both aspects through the implementation of self aligning LIGA (German acronym for lithographie, galvanoformung, abformung, or x ray lithography, electroplating, and molding) optomechanics with high volume plastic injection molded and off-the-shelf glass optics is presented. This zero alignment strategy has been incorporated into a miniature high numerical aperture (NA = 1.0 W) microscope objective for a fiber confocal reflectance microscope. Tight alignment tolerances of less than 10 microm are maintained for all components that reside inside of a small 9 gauge diameter hypodermic tubing. A prototype system has been tested using the slanted edge modulation transfer function technique and demonstrated to have a Strehl ratio of 0.71. This universal technology is now being developed for smaller, needle-sized imaging systems and other portable point-of-care diagnostic instruments. PMID- 19543345 TI - Design and analysis of metal-dielectric nonpolarizing beam splitters in a glass cube. AB - A novel design of a 25-layer metal-dielectric nonpolarizing beam splitter in a cube is proposed by use of the optimization method and is theoretically investigated. The simulations of the reflectance and differential phases induced by reflection and transmission are presented. The simulation results reveal that both the amplitude and the phase characteristics of the nonpolarizing beam splitter could realize the design targets, the differences between the simulated and the target reflectance of 50% are less than 2%, and the differential phases are less than 3 degrees in the range of 530 nm-570 nm for both p and s components. PMID- 19543346 TI - Design of a relay lens with telecentricity in a holographic recording system. AB - We reveal a doubly telecentric Fourier 4f relay for a holographic recording system. These lenses are made of Zeonex and their aspherical surfaces were formed by a diamond-turning machining. The total stroke of the system is 244.69 mm, including six lenses. The relay system provides a small distortion and the root mean-square error of the wavefront within 1/4lambda (lambda = 532 nm). In addition, we describe the optical behavior of the designed system and present our experiences at manufacturing aspheric lens prototypes. PMID- 19543347 TI - Rigid and high-numerical-aperture two-photon fluorescence endoscope. AB - We present a rigid miniaturized optical system block fiber-optic two-photon endoscope based on a compact two-axis piezo scanner system and a miniature high (0.65) NA GRIN lens objective. The optical system is scanned as a whole by a piezo scanner allowing always an on-axis beam irradiation of the optical system. A photonic crystal fiber is used for excitation and ultrashort laser pulses can be delivered with typical power up to 100 mW at 800 nm. Two-photon fluorescence signal is collected by the use of a multimode fiber. Lateral resolution values for the system were experimentally measured to be 0.67 microm vertically and 1.08 microm horizontally. Axial resolution was found to be 5.8 microm. The endoscope is highly flexible and controllable in terms of time acquisition, resolution, and magnification. Fluorescence images were acquired over a 420 microm x 420 microm field of view. Results presented here demonstrate the ability of the system to resolve subcellular details and the potential of the technology for in vivo applications. PMID- 19543349 TI - Backward discrete wave field propagation modeling as an inverse problem: toward perfect reconstruction of wave field distributions. AB - We consider reconstruction of a wave field distribution in an input/object plane from data in an output/diffraction (sensor) plane. We provide digital modeling both for the forward and backward wave field propagation. A novel algebraic matrix form of the discrete diffraction transform (DDT) originated in Katkovnik et al. [Appl. Opt. 47, 3481 (2008)] is proposed for the forward modeling that is aliasing free and precise for pixelwise invariant object and sensor plane distributions. This "matrix DDT" is a base for formalization of the object wave field reconstruction (backward propagation) as an inverse problem. The transfer matrices of the matrix DDT are used for calculations as well as for the analysis of conditions when the perfect reconstruction of the object wave field distribution is possible. We show by simulation that the developed inverse propagation algorithm demonstrates an improved accuracy as compared with the standard convolutional and discrete Fresnel transform algorithms. PMID- 19543348 TI - High-speed linear detection time domain optical coherence tomography with reflective grating-generated spatial reference delay. AB - We developed a high-speed linear detection time-domain optical coherence tomography (OCT) technique that detected a reflective grating-generated spatial optical delay in the reference arm using a line scan camera during probe-beam scanning. Using an InGaAs line scan camera (512 pixels) operating at 47,000 lines/s, the calculation of the absolute value of the difference between two sequential lines can be approximately displayed as cross-sectional images with 500 lateral pixels at 94 frames/s . After data acquisition, we performed postprocessing that involves a Hilbert transform to improve the image quality of an OCT image. Our OCT system was successfully used to image a human finger in vivo with 93 dB sensitivity. PMID- 19543350 TI - Reduction of gain depletion and saturation on a Brillouin-erbium fiber laser utilizing a Brillouin pump preamplification technique. AB - This paper presents the characteristics of a multiwavelength L-band Brillouin erbium comb fiber laser with a preamplified Brillouin pump (BP) power technique at low pumping powers. The issue of erbium-doped fiber gain depletion and Brillouin gain saturation are resolved by the proposed structure. For long single mode fiber length, the Stokes line emission occurs at low pumping powers because of the high strength of spontaneous Brillouin scattering, which provides a strong seed for coherent regenerative amplification of the Stokes line in the laser cavity. The laser structure achieves a low threshold power of 17 mW and is able to produce high number of output channels at low pumping powers. We experimentally show that the fiber laser structure can produce up to 37 channels at 55 and 0.045 mW of 1480 nm pump and BP powers, respectively. PMID- 19543351 TI - Directly photoinscribed refractive index change and Bragg gratings in Ohara WMS 15 glass ceramic. AB - We inscribed thick volume gratings in WMS-15 glass ceramic by ultraviolet light at 193 and 248 nm. Unlike earlier work in ceramic materials, the inscription process modified the optical properties of the material without the need for any additional chemical or thermal processing. Experimental evidence from measurements of grating growth, thermal annealing, and spectral absorption indicates that two distinct physical mechanisms are responsible for the grating formation. Weak, easily thermally bleached gratings resulted from exposure fluences below 0.3 kJ/cm2. Optical absorption measurements suggest that these low fluence gratings are predominantly absorption gratings. More thermally stable gratings, found to be refractive index gratings with unsaturated refractive index modulation amplitude as large as 6 x 10(-5) were formed at cumulative fluences of 1 kJ/cm2 and above. PMID- 19543352 TI - Optical wave fields with lateral and longitudinal periodicity. AB - The propagation of stationary wave fields that exhibit simultaneously lateral and longitudinal periodicity is investigated. As a model, we use a Fabry-Perot resonator with periodically structured mirrors under monochromatic plane wave illumination. The resonator leads to a longitudinal periodicity, the grating mirrors to a lateral periodicity. The angular spectrum of the transmitted wave field is given as the product of two terms, one related to the lateral, the other to the longitudinal properties. Its modal structure can vary significantly depending on the ratio of the lateral and longitudinal periods and the reflectivity of the resonator's mirrors. For example, it is possible to generate bandgap behavior despite the fact that the periods may be significantly larger than the wavelength. The results of this investigation apply to the design of phase-coupled array resonators and multiplexers. PMID- 19543353 TI - Liquid crystal panel for high efficiency barrier type autostereoscopic three dimensional displays. AB - An autostereoscopic display with parallax barrier attached onto a liquid crystal panel suffers from the trade-off between brightness and crosstalk. One approach for making improvement by modifying the layout of light blocking components, such as thin film transistor, storage capacitor, and protrusion, in the liquid crystal pixel has been proposed. Ray tracing simulation shows that the aperture of the slanted barrier can be significantly increased, hence increasing efficiency, while keeping the same crosstalk level if those light blocking components can be shifted to the corner of the pixel. A six-view 2.83 in. (7.19 cm) prototype has shown improvement on both brightness and crosstalk compared to its counterpart using a traditional liquid crystal panel, which demonstrates an effective approach for a high-efficiency barrier-type autostereoscopic 3D display with a liquid crystal panel. PMID- 19543354 TI - Single- and dual-wavelength switchable erbium-doped fiber ring laser based on intracavity polarization selective tilted fiber gratings. AB - We propose and demonstrate a single- and dual-wavelength switchable erbium-doped fiber laser (EDFL) by utilizing intracavity polarization selective filters based on tilted fiber gratings (TFGs). In the cavity, one 45 degrees TFG functions as an in-fiber polarizer and the other 77 degrees TFG is used as a fiber polarization dependent loss (PDL) filter. The combined polarization effect from these two TFGs enables the laser to switch between the single- and the dual wavelength operation with a single-polarization state at room temperature. The laser output at each wavelength shows an optical signal-to-noise ratio (OSNR) of >60 dB, a side mode suppression ratio (SMSR) of >50 dB, and a polarization extinction ratio of approximately 35 dB. The proposed EDFL can give stable output under laboratory conditions. PMID- 19543355 TI - Acoustic effects of metal vapor lasers. AB - Some fluctuations in the output power of a copper vapor laser with a 16 mm bore were recorded by varying the excitation frequency from 13 to 33 kHz. The effect arises from the laser tube, which performs both as an optical and an acoustic resonator at the acoustic resonant frequencies. It is shown that a similar effect occurs in other metal vapor and copper halide lasers as well. PMID- 19543356 TI - Bragg cell laser intensity modulation: effect on laser Doppler velocimetry measurements. AB - In most laser Doppler velocimetry (LDV) systems, the frequency of one of the two laser beams that intersect to create the probe volume is shifted with an acousto optic element. It is shown here that Bragg shifting can impose a problematic fluctuation in intensity on the frequency-shifted beam, producing spurious velocity measurements. This fluctuation occurs at twice the Bragg cell frequency, and its relative amplitude to the time average intensity is a function of the ratio of the laser beam diameter to the Bragg cell acoustic wavelength. A physical model and a configuration procedure to minimize adverse effects of the intensity modulations are presented. PMID- 19543357 TI - Separate recording of rationally related vibration frequencies using digital stroboscopic holographic interferometry. AB - A method for separate recording of rationally related vibration frequencies is presented. To record and measure the mode shape of vibrations, a synchronized stroboscopic CCD camera is used. Synchronization and control of the camera acquisition for recording stroboscopic holographic sequence has been realized. The phase for different states of the object vibration is calculated using the Fourier-transform method. Experimental results are presented, and the advantages and disadvantages of the proposed method are discussed. PMID- 19543359 TI - Simple two-layer reflectance model for biological tissue applications. AB - A two-layer tissue diffuse reflectance model is described. The model is based on a simple one-layer model that we have recently developed and successfully applied to the analysis of in vivo skin reflectance. The model, which is specifically designed for use with a fiber optic probe, has as its main features simplicity and ease of application, and it is capable of estimating the thickness and the absorption coefficient of a superficial absorbing and scattering layer. Both of these parameters are of great interest for the noninvasive study of epithelial biological tissues. The validity range and accuracy of the model are tested on tissue phantoms in both the forward and inverse modes of application. PMID- 19543358 TI - Minimum refractometrically detectable concentrations of single atmospheric gases and simple mixtures with a Sagnac interferometer. AB - The minimum quantities of the nine most abundant, isolated, atmospheric gases that are detectable with a refractometer are calculated. An examination of the applicability of refractometric techniques for detecting and analyzing gaseous mixtures is discussed and a comparison made against other established techniques. Traditionally, most gas analysis performed with an interferometer is in determining the dispersion or refractivity of a known sample, presented here is the inverse approach, where refractivities are measured to determine the concentrations of particular species within a gas. The method, and experimental results for determining the minimum quantities of a particular species detectable in a mixture has been explored, as well as the complications, such as the indistinguishability of dynamic polarizabilities of different gases and the subsequent demands for accurate pressure and fringe measurements of using interferometric techniques. It is shown that the concentration of a single (isolated) gas, in units of number density, can be determined to within approximately 1-10 x 10(18) m(-3), and a mixture of the three most abundant gases, N2, O2 and Ar, to within 3.4 x 10(4) parts in 10(6) (ppm) when a minimum detectable fringe shift of lambda/100 is assumed. PMID- 19543361 TI - Bulk optical damage thresholds for doped and undoped, crystalline and ceramic yttrium aluminum garnet. AB - We measured the bulk optical damage thresholds of pure and Nd-doped ceramic yttrium aluminum garnet (YAG), and of pure, Nd-doped, Cr-doped, and Yb-doped crystalline YAG. We used 9.9 ns, 1064 nm, single-longitudinal mode, TEM00 pulses, to determine that the breakdown thresholds are deterministic, with multiple-pulse thresholds ranging from 1.1 to 2.2 kJ/cm2. PMID- 19543360 TI - Interpolated local model fitting method for accurate and fast single-shot surface profiling. AB - The local model fitting (LMF) method is a useful single-shot surface profiling algorithm based on spatial carrier frequency fringe patterns. The measurement principle of the LMF method relies on the assumption that the target surface is locally flat. In this paper, we first analyze the measurement error of the LMF method caused by violation of the locally flat assumption. More specifically, we theoretically prove that the measurement error is zero at fringe intensity extrema in an interference pattern even when the locally flat assumption is violated. Based on this theoretical finding, we propose a new surface profiling method called the interpolated LMF (iLMF) algorithm, which is more accurate and computationally efficient than the original LMF method. The practical usefulness of the iLMF method is shown through experiments. PMID- 19543362 TI - Eavesdropping in chaotic optical communication using the feedback length of an external-cavity laser as a key. AB - An external-cavity laser (ECL) operating in a chaotic state is usually used in a chaotic optical secure communication system and its feedback length (FL) is often regarded as an additional key. Our analyses show that an eavesdropper's (Eve) laser can synchronize with a transmitter (Alice) without any knowledge of the FL by simply increasing the injection strength. A sequence of a 1 Gbit/s nonreturn to-zero message encoded by the FL as the key is successfully eavesdropped. The reason for the synchronization deviation between Alice's and Eve's lasers is given. Our results indicate that the FL as a key cannot enhance the security of chaotic optical communication using long-ECLs. PMID- 19543363 TI - High G+C Content of Herpes Simplex Virus DNA: Proposed Role in Protection Against Retrotransposon Insertion. AB - The herpes simplex virus dsDNA genome is distinguished by an unusually high G+C nucleotide content. HSV-1 and HSV-2, for instance, have GC contents of 68% and 70% respectively, while that of the host (human) genome is 41%. To determine how GC content varies with genome location, GC content was measured separately in coding and intergenic regions of HSV-1 DNA. The results showed that the 75 genes constitute a uniform population with a mean GC content of 66.9 +/- 4.1%. In contrast, intergenic regions were found in two non-overlapping populations, one with a mean GC content (69.3 +/- 4.6% n=32) similar to the coding regions and another where the GC content is lower (56.0 +/- 4.9 n=30). Compared to other regions of the genome, intergenic regions with reduced GC content were found to be enriched in local GC minima, CACACA sequences and a primary target sequence (TTAAAA) for retrotransposition events. The results are interpreted to suggest that a high GC content is part of the way HSV-1 protects its genes from invasion by mobile genetic elements active during cell differentiation in the nervous system. PMID- 19543364 TI - The maternal-effect gene cellular island encodes aurora B kinase and is essential for furrow formation in the early zebrafish embryo. AB - Females homozygous for a mutation in cellular island (cei) produce embryos with defects in cytokinesis during early development. Analysis of the cytoskeletal events associated with furrow formation reveal that these defects include a general delay in furrow initiation as well as a complete failure to form furrow associated structures in distal regions of the blastodisc. A linkage mapping based candidate gene approach, including transgenic rescue, shows that cei encodes the zebrafish Aurora B kinase homologue. Genetic complementation analysis between the cei mutation and aurB zygotic lethal mutations corroborate gene assignment and reveal a complex nature of the maternal-effect cei allele, which appears to preferentially affect a function important for cytokinesis in the early blastomeres. Surprisingly, in cei mutant embryos a short yet otherwise normal furrow forms in the center of the blastodisc. Furrow formation is absent throughout the width of the blastodisc in cei mutant embryos additionally mutant for futile cycle, which lack a spindle apparatus, showing that the residual furrow signal present in cei mutants is derived from the mitotic spindle. Our analysis suggests that partially redundant signals derived from the spindle and astral apparatus mediate furrow formation in medial and distal regions of the early embryonic blastomeres, respectively, possibly as a spatial specialization to achieve furrow formation in these large cells. In addition, our data also suggest a role for Cei/AurB function in the reorganization of the furrow associated microtubules in both early cleavage- and somite-stage embryos. In accordance with the requirement for cei/aurB in furrow induction in the early cleavage embryo, germ plasm recruitment to the forming furrow is also affected in embryos lacking normal cei/aurB function. PMID- 19543365 TI - Peroxiredoxin Tsa1 is the key peroxidase suppressing genome instability and protecting against cell death in Saccharomyces cerevisiae. AB - Peroxiredoxins (Prxs) constitute a family of thiol-specific peroxidases that utilize cysteine (Cys) as the primary site of oxidation during the reduction of peroxides. To gain more insight into the physiological role of the five Prxs in budding yeast Saccharomyces cerevisiae, we performed a comparative study and found that Tsa1 was distinguished from the other Prxs in that by itself it played a key role in maintaining genome stability and in sustaining aerobic viability of rad51 mutants that are deficient in recombinational repair. Tsa2 and Dot5 played minor but distinct roles in suppressing the accumulation of mutations in cooperation with Tsa1. Tsa2 was capable of largely complementing the absence of Tsa1 when expressed under the control of the Tsa1 promoter. The presence of peroxidatic cysteine (Cys(47)) was essential for Tsa1 activity, while Tsa1(C170S) lacking the resolving Cys was partially functional. In the absence of Tsa1 activity (tsa1 or tsa1(CCS) lacking the peroxidatic and resolving Cys) and recombinational repair (rad51), dying cells displayed irregular cell size/shape, abnormal cell cycle progression, and significant increase of phosphatidylserine externalization, an early marker of apoptosis-like cell death. The tsa1(CCS) rad51- or tsa1 rad51-induced cell death did not depend on the caspase Yca1 and Ste20 kinase, while the absence of the checkpoint protein Rad9 accelerated the cell death processes. These results indicate that the peroxiredoxin Tsa1, in cooperation with appropriate DNA repair and checkpoint mechanisms, acts to protect S. cerevisiae cells against toxic levels of DNA damage that occur during aerobic growth. PMID- 19543367 TI - Avoiding dangerous missense: thermophiles display especially low mutation rates. AB - Rates of spontaneous mutation have been estimated under optimal growth conditions for a variety of DNA-based microbes, including viruses, bacteria, and eukaryotes. When expressed as genomic mutation rates, most of the values were in the vicinity of 0.003-0.004 with a range of less than two-fold. Because the genome sizes varied by roughly 10(4)-fold, the mutation rates per average base pair varied inversely by a similar factor. Even though the commonality of the observed genomic rates remains unexplained, it implies that mutation rates in unstressed microbes reach values that can be finely tuned by evolution. An insight originating in the 1920s and maturing in the 1960s proposed that the genomic mutation rate would reflect a balance between the deleterious effect of the average mutation and the cost of further reducing the mutation rate. If this view is correct, then increasing the deleterious impact of the average mutation should be countered by reducing the genomic mutation rate. It is a common observation that many neutral or nearly neutral mutations become strongly deleterious at higher temperatures, in which case they are called temperature-sensitive mutations. Recently, the kinds and rates of spontaneous mutations were described for two microbial thermophiles, a bacterium and an archaeon. Using an updated method to extrapolate from mutation-reporter genes to whole genomes reveals that the rate of base substitutions is substantially lower in these two thermophiles than in mesophiles. This result provides the first experimental support for the concept of an evolved balance between the total genomic impact of mutations and the cost of further reducing the basal mutation rate. PMID- 19543366 TI - A network of conserved damage survival pathways revealed by a genomic RNAi screen. AB - Damage initiates a pleiotropic cellular response aimed at cellular survival when appropriate. To identify genes required for damage survival, we used a cell-based RNAi screen against the Drosophila genome and the alkylating agent methyl methanesulphonate (MMS). Similar studies performed in other model organisms report that damage response may involve pleiotropic cellular processes other than the central DNA repair components, yet an intuitive systems level view of the cellular components required for damage survival, their interrelationship, and contextual importance has been lacking. Further, by comparing data from different model organisms, identification of conserved and presumably core survival components should be forthcoming. We identified 307 genes, representing 13 signaling, metabolic, or enzymatic pathways, affecting cellular survival of MMS induced damage. As expected, the majority of these pathways are involved in DNA repair; however, several pathways with more diverse biological functions were also identified, including the TOR pathway, transcription, translation, proteasome, glutathione synthesis, ATP synthesis, and Notch signaling, and these were equally important in damage survival. Comparison with genomic screen data from Saccharomyces cerevisiae revealed no overlap enrichment of individual genes between the species, but a conservation of the pathways. To demonstrate the functional conservation of pathways, five were tested in Drosophila and mouse cells, with each pathway responding to alkylation damage in both species. Using the protein interactome, a significant level of connectivity was observed between Drosophila MMS survival proteins, suggesting a higher order relationship. This connectivity was dramatically improved by incorporating the components of the 13 identified pathways within the network. Grouping proteins into "pathway nodes" qualitatively improved the interactome organization, revealing a highly organized "MMS survival network." We conclude that identification of pathways can facilitate comparative biology analysis when direct gene/orthologue comparisons fail. A biologically intuitive, highly interconnected MMS survival network was revealed after we incorporated pathway data in our interactome analysis. PMID- 19543368 TI - Disease-causing 7.4 kb cis-regulatory deletion disrupting conserved non-coding sequences and their interaction with the FOXL2 promotor: implications for mutation screening. AB - To date, the contribution of disrupted potentially cis-regulatory conserved non coding sequences (CNCs) to human disease is most likely underestimated, as no systematic screens for putative deleterious variations in CNCs have been conducted. As a model for monogenic disease we studied the involvement of genetic changes of CNCs in the cis-regulatory domain of FOXL2 in blepharophimosis syndrome (BPES). Fifty-seven molecularly unsolved BPES patients underwent high resolution copy number screening and targeted sequencing of CNCs. Apart from three larger distant deletions, a de novo deletion as small as 7.4 kb was found at 283 kb 5' to FOXL2. The deletion appeared to be triggered by an H-DNA-induced double-stranded break (DSB). In addition, it disrupts a novel long non-coding RNA (ncRNA) PISRT1 and 8 CNCs. The regulatory potential of the deleted CNCs was substantiated by in vitro luciferase assays. Interestingly, Chromosome Conformation Capture (3C) of a 625 kb region surrounding FOXL2 in expressing cellular systems revealed physical interactions of three upstream fragments and the FOXL2 core promoter. Importantly, one of these contains the 7.4 kb deleted fragment. Overall, this study revealed the smallest distant deletion causing monogenic disease and impacts upon the concept of mutation screening in human disease and developmental disorders in particular. PMID- 19543369 TI - Comprehensive linkage and association analyses identify haplotype, near to the TNFSF15 gene, significantly associated with spondyloarthritis. AB - Spondyloarthritis (SpA) is a chronic inflammatory disorder with a strong genetic predisposition dominated by the role of HLA-B27. However, the contribution of other genes to the disease susceptibility has been clearly demonstrated. We previously reported significant evidence of linkage of SpA to chromosome 9q31-34. The current study aimed to characterize this locus, named SPA2. First, we performed a fine linkage mapping of SPA2 (24 cM) with 28 microsatellite markers in 149 multiplex families, which allowed us to reduce the area of investigation to an 18 cM (13 Mb) locus delimited by the markers D9S279 and D9S112. Second, we constructed a linkage disequilibrium (LD) map of this region with 1,536 tag single-nucleotide polymorphisms (SNPs) in 136 families (263 patients). The association was assessed using a transmission disequilibrium test. One tag SNP, rs4979459, yielded a significant P-value (4.9 x 10(-5)). Third, we performed an extension association study with rs4979459 and 30 surrounding SNPs in LD with it, in 287 families (668 patients), and in a sample of 139 cases and 163 controls. Strong association was observed in both familial and case/control datasets for several SNPs. In the replication study, carried with 8 SNPs in an independent sample of 232 cases and 149 controls, one SNP, rs6478105, yielded a nominal P value<3 x 10(-2). Pooled case/control study (371 cases and 312 controls) as well as combined analysis of extension and replication data showed very significant association (P<5 x 10(-4)) for 6 of the 8 latter markers (rs7849556, rs10817669, rs10759734, rs6478105, rs10982396, and rs10733612). Finally, haplotype association investigations identified a strongly associated haplotype (P<8.8 x 10(-5)) consisting of these 6 SNPs and located in the direct vicinity of the TNFSF15 gene. In conclusion, we have identified within the SPA2 locus a haplotype strongly associated with predisposition to SpA which is located near to TNFSF15, one of the major candidate genes in this region. PMID- 19543370 TI - Sensitive detection of chromosomal segments of distinct ancestry in admixed populations. AB - Identifying the ancestry of chromosomal segments of distinct ancestry has a wide range of applications from disease mapping to learning about history. Most methods require the use of unlinked markers; but, using all markers from genome wide scanning arrays, it should in principle be possible to infer the ancestry of even very small segments with exquisite accuracy. We describe a method, HAPMIX, which employs an explicit population genetic model to perform such local ancestry inference based on fine-scale variation data. We show that HAPMIX outperforms other methods, and we explore its utility for inferring ancestry, learning about ancestral populations, and inferring dates of admixture. We validate the method empirically by applying it to populations that have experienced recent and ancient admixture: 935 African Americans from the United States and 29 Mozabites from North Africa. HAPMIX will be of particular utility for mapping disease genes in recently admixed populations, as its accurate estimates of local ancestry permit admixture and case-control association signals to be combined, enabling more powerful tests of association than with either signal alone. PMID- 19543371 TI - Genetic variation of promoter sequence modulates XBP1 expression and genetic risk for vitiligo. AB - Our previous genome-wide linkage analysis identified a susceptibility locus for generalized vitiligo on 22q12. To search for susceptibility genes within the locus, we investigated a biological candidate gene, X-box binding protein 1(XBP1). First, we sequenced all the exons, exon-intron boundaries as well as some 5' and 3' flanking sequences of XBP1 in 319 cases and 294 controls of Chinese Hans. Of the 8 common variants identified, the significant association was observed at rs2269577 (p_(trend) = 0.007, OR = 1.36, 95% CI = 1.09-1.71), a putative regulatory polymorphism within the promoter region of XBP1. We then sequenced the variant in an additional 365 cases and 404 controls and found supporting evidence for the association (p_(trend) = 0.008, OR = 1.31, 95% CI = 1.07-1.59). To further validate the association, we genotyped the variant in another independent sample of 1,402 cases and 1,288 controls, including 94 parent child trios, and confirmed the association by both case-control analysis (p_(trend) = 0.003, OR = 1.18, 95% CI = 1.06-1.32) and the family-based transmission disequilibrium test (TDT, p = 0.005, OR = 1.93, 95% CI = 1.21-3.07). The analysis of the combined 2,086 cases and 1,986 controls provided highly significant evidence for the association (p_(trend) = 2.94x10(-6), OR = 1.23, 95% CI = 1.13-1.35). Furthermore, we also found suggestive epistatic effect between rs2269577 and HLA-DRB1*07 allele on the development of vitiligo (p = 0.033). Our subsequent functional study showed that the risk-associated C allele of rs2269577 had a stronger promoter activity than the non-risk G allele, and there was an elevated expression of XBP1 in the lesional skins of patients carrying the risk associated C allele. Therefore, our study has demonstrated that the transcriptional modulation of XBP1 expression by a germ-line regulatory polymorphism has an impact on the development of vitiligo. PMID- 19543372 TI - Genome-wide identification of alternative splice forms down-regulated by nonsense mediated mRNA decay in Drosophila. AB - Alternative mRNA splicing adds a layer of regulation to the expression of thousands of genes in Drosophila melanogaster. Not all alternative splicing results in functional protein; it can also yield mRNA isoforms with premature stop codons that are degraded by the nonsense-mediated mRNA decay (NMD) pathway. This coupling of alternative splicing and NMD provides a mechanism for gene regulation that is highly conserved in mammals. NMD is also active in Drosophila, but its effect on the repertoire of alternative splice forms has been unknown, as has the mechanism by which it recognizes targets. Here, we have employed a custom splicing-sensitive microarray to globally measure the effect of alternative mRNA processing and NMD on Drosophila gene expression. We have developed a new algorithm to infer the expression change of each mRNA isoform of a gene based on the microarray measurements. This method is of general utility for interpreting splicing-sensitive microarrays and high-throughput sequence data. Using this approach, we have identified a high-confidence set of 45 genes where NMD has a differential effect on distinct alternative isoforms, including numerous RNA binding and ribosomal proteins. Coupled alternative splicing and NMD decrease expression of these genes, which may in turn have a downstream effect on expression of other genes. The NMD-affected genes are enriched for roles in translation and mitosis, perhaps underlying the previously observed role of NMD factors in cell cycle progression. Our results have general implications for understanding the NMD mechanism in fly. Most notably, we found that the NMD target mRNAs had significantly longer 3' untranslated regions (UTRs) than the nontarget isoforms of the same genes, supporting a role for 3' UTR length in the recognition of NMD targets in fly. PMID- 19543373 TI - A flexible and accurate genotype imputation method for the next generation of genome-wide association studies. AB - Genotype imputation methods are now being widely used in the analysis of genome wide association studies. Most imputation analyses to date have used the HapMap as a reference dataset, but new reference panels (such as controls genotyped on multiple SNP chips and densely typed samples from the 1,000 Genomes Project) will soon allow a broader range of SNPs to be imputed with higher accuracy, thereby increasing power. We describe a genotype imputation method (IMPUTE version 2) that is designed to address the challenges presented by these new datasets. The main innovation of our approach is a flexible modelling framework that increases accuracy and combines information across multiple reference panels while remaining computationally feasible. We find that IMPUTE v2 attains higher accuracy than other methods when the HapMap provides the sole reference panel, but that the size of the panel constrains the improvements that can be made. We also find that imputation accuracy can be greatly enhanced by expanding the reference panel to contain thousands of chromosomes and that IMPUTE v2 outperforms other methods in this setting at both rare and common SNPs, with overall error rates that are 15%-20% lower than those of the closest competing method. One particularly challenging aspect of next-generation association studies is to integrate information across multiple reference panels genotyped on different sets of SNPs; we show that our approach to this problem has practical advantages over other suggested solutions. PMID- 19543374 TI - A tether for Woronin body inheritance is associated with evolutionary variation in organelle positioning. AB - Eukaryotic organelles evolve to support the lifestyle of evolutionarily related organisms. In the fungi, filamentous Ascomycetes possess dense-core organelles called Woronin bodies (WBs). These organelles originate from peroxisomes and perform an adaptive function to seal septal pores in response to cellular wounding. Here, we identify Leashin, an organellar tether required for WB inheritance, and associate it with evolutionary variation in the subcellular pattern of WB distribution. In Neurospora, the leashin (lah) locus encodes two related adjacent genes. N-terminal sequences of LAH-1 bind WBs via the WB specific membrane protein WSC, and C-terminal sequences are required for WB inheritance by cell cortex association. LAH-2 is localized to the hyphal apex and septal pore rim and plays a role in colonial growth. In most species, WBs are tethered directly to the pore rim, however, Neurospora and relatives have evolved a delocalized pattern of cortex association. Using a new method for the construction of chromosomally encoded fusion proteins, marker fusion tagging (MFT), we show that a LAH-1/LAH-2 fusion can reproduce the ancestral pattern in Neurospora. Our results identify the link between the WB and cell cortex and suggest that splitting of leashin played a key role in the adaptive evolution of organelle localization. PMID- 19543376 TI - Selective processing and metabolism of disease-causing mutant prion proteins. AB - Prion diseases are fatal neurodegenerative disorders caused by aberrant metabolism of the cellular prion protein (PrP(C)). In genetic forms of these diseases, mutations in the globular C-terminal domain are hypothesized to favor the spontaneous generation of misfolded PrP conformers (including the transmissible PrP(Sc) form) that trigger downstream pathways leading to neuronal death. A mechanistic understanding of these diseases therefore requires knowledge of the quality control pathways that recognize and degrade aberrant PrPs. Here, we present comparative analyses of the biosynthesis, trafficking, and metabolism of a panel of genetic disease-causing prion protein mutants in the C-terminal domain. Using quantitative imaging and biochemistry, we identify a misfolded subpopulation of each mutant PrP characterized by relative detergent insolubility, inaccessibility to the cell surface, and incomplete glycan modifications. The misfolded populations of mutant PrPs were neither recognized by ER quality control pathways nor routed to ER-associated degradation despite demonstrable misfolding in the ER. Instead, mutant PrPs trafficked to the Golgi, from where the misfolded subpopulation was selectively trafficked for degradation in acidic compartments. Surprisingly, selective re-routing was dependent not only on a mutant globular domain, but on an additional lysine-based motif in the highly conserved unstructured N-terminus. These results define a specific trafficking and degradation pathway shared by many disease-causing PrP mutants. As the acidic lysosomal environment has been implicated in facilitating the conversion of PrP(C) to PrP(Sc), our identification of a mutant-selective trafficking pathway to this compartment may provide a cell biological basis for spontaneous generation of PrP(Sc) in familial prion disease. PMID- 19543375 TI - Vector transmission of leishmania abrogates vaccine-induced protective immunity. AB - Numerous experimental vaccines have been developed to protect against the cutaneous and visceral forms of leishmaniasis caused by infection with the obligate intracellular protozoan Leishmania, but a human vaccine still does not exist. Remarkably, the efficacy of anti-Leishmania vaccines has never been fully evaluated under experimental conditions following natural vector transmission by infected sand fly bite. The only immunization strategy known to protect humans against natural exposure is "leishmanization," in which viable L. major parasites are intentionally inoculated into a selected site in the skin. We employed mice with healed L. major infections to mimic leishmanization, and found tissue seeking, cytokine-producing CD4+ T cells specific for Leishmania at the site of challenge by infected sand fly bite within 24 hours, and these mice were highly resistant to sand fly transmitted infection. In contrast, mice vaccinated with a killed vaccine comprised of autoclaved L. major antigen (ALM)+CpG oligodeoxynucleotides that protected against needle inoculation of parasites, showed delayed expression of protective immunity and failed to protect against infected sand fly challenge. Two-photon intra-vital microscopy and flow cytometric analysis revealed that sand fly, but not needle challenge, resulted in the maintenance of a localized neutrophilic response at the inoculation site, and removal of neutrophils following vector transmission led to increased parasite specific immune responses and promoted the efficacy of the killed vaccine. These observations identify the critical immunological factors influencing vaccine efficacy following natural transmission of Leishmania. PMID- 19543377 TI - Identification of yeast transcriptional regulation networks using multivariate random forests. AB - The recent availability of whole-genome scale data sets that investigate complementary and diverse aspects of transcriptional regulation has spawned an increased need for new and effective computational approaches to analyze and integrate these large scale assays. Here, we propose a novel algorithm, based on random forest methodology, to relate gene expression (as derived from expression microarrays) to sequence features residing in gene promoters (as derived from DNA motif data) and transcription factor binding to gene promoters (as derived from tiling microarrays). We extend the random forest approach to model a multivariate response as represented, for example, by time-course gene expression measures. An analysis of the multivariate random forest output reveals complex regulatory networks, which consist of cohesive, condition-dependent regulatory cliques. Each regulatory clique features homogeneous gene expression profiles and common motifs or synergistic motif groups. We apply our method to several yeast physiological processes: cell cycle, sporulation, and various stress conditions. Our technique displays excellent performance with regard to identifying known regulatory motifs, including high order interactions. In addition, we present evidence of the existence of an alternative MCB-binding pathway, which we confirm using data from two independent cell cycle studies and two other physioloigical processes. Finally, we have uncovered elaborate transcription regulation refinement mechanisms involving PAC and mRRPE motifs that govern essential rRNA processing. These include intriguing instances of differing motif dosages and differing combinatorial motif control that promote regulatory specificity in rRNA metabolism under differing physiological processes. PMID- 19543379 TI - Intravenous inoculation of a bat-associated rabies virus causes lethal encephalopathy in mice through invasion of the brain via neurosecretory hypothalamic fibers. AB - The majority of rabies virus (RV) infections are caused by bites or scratches from rabid carnivores or bats. Usually, RV utilizes the retrograde transport within the neuronal network to spread from the infection site to the central nervous system (CNS) where it replicates in neuronal somata and infects other neurons via trans-synaptic spread. We speculate that in addition to the neuronal transport of the virus, hematogenous spread from the site of infection directly to the brain after accidental spill over into the vascular system might represent an alternative way for RV to invade the CNS. So far, it is unknown whether hematogenous spread has any relevance in RV pathogenesis. To determine whether certain RV variants might have the capacity to invade the CNS from the periphery via hematogenous spread, we infected mice either intramuscularly (i.m.) or intravenously (i.v.) with the dog-associated RV DOG4 or the silver-haired bat associated RV SB. In addition to monitoring the progression of clinical signs of rabies we used immunohistochemistry and quantitative reverse transcription polymerase chain reaction (qRT-PCR) to follow the spread of the virus from the infection site to the brain. In contrast to i.m. infection where both variants caused a lethal encephalopathy, only i.v. infection with SB resulted in the development of a lethal infection. While qRT-PCR did not reveal major differences in virus loads in spinal cord or brain at different times after i.m. or i.v. infection of SB, immunohistochemical analysis showed that only i.v. administered SB directly infected the forebrain. The earliest affected regions were those hypothalamic nuclei, which are connected by neurosecretory fibers to the circumventricular organs neurohypophysis and median eminence. Our data suggest that hematogenous spread of SB can lead to a fatal encephalopathy through direct retrograde invasion of the CNS at the neurovascular interface of the hypothalamus hypophysis system. This alternative mode of virus spread has implications for the post exposure prophylaxis of rabies, particularly with silver-haired bat associated RV. PMID- 19543378 TI - Connecting quorum sensing, c-di-GMP, pel polysaccharide, and biofilm formation in Pseudomonas aeruginosa through tyrosine phosphatase TpbA (PA3885). AB - With the opportunistic pathogen Pseudomonas aeruginosa, quorum sensing based on homoserine lactones was found to influence biofilm formation. Here we discern a mechanism by which quorum sensing controls biofilm formation by screening 5850 transposon mutants of P. aeruginosa PA14 for altered biofilm formation. This screen identified the PA3885 mutant, which had 147-fold more biofilm than the wild-type strain. Loss of PA3885 decreased swimming, abolished swarming, and increased attachment, although this did not affect production of rhamnolipids. The PA3885 mutant also had a wrinkly colony phenotype, formed pronounced pellicles, had substantially more aggregation, and had 28-fold more exopolysaccharide production. Expression of PA3885 in trans reduced biofilm formation and abolished aggregation. Whole transcriptome analysis showed that loss of PA3885 activated expression of the pel locus, an operon that encodes for the synthesis of extracellular matrix polysaccharide. Genetic screening identified that loss of PelABDEG and the PA1120 protein (which contains a GGDEF motif) suppressed the phenotypes of the PA3885 mutant, suggesting that the function of the PA3885 protein is to regulate 3,5-cyclic diguanylic acid (c-di GMP) concentrations as a phosphatase since c-di-GMP enhances biofilm formation by activating PelD, and c-di-GMP inhibits swarming. Loss of PA3885 protein increased cellular c-di-GMP concentrations; hence, PA3885 protein is a negative regulator of c-di-GMP production. Purified PA3885 protein has phosphatase activity against phosphotyrosine peptides and is translocated to the periplasm. Las-mediated quorum sensing positively regulates expression of the PA3885 gene. These results show that the PA3885 protein responds to AHL signals and likely dephosphorylates PA1120, which leads to reduced c-di-GMP production. This inhibits matrix exopolysaccharide formation, which leads to reduced biofilm formation; hence, we provide a mechanism for quorum sensing control of biofilm formation through the pel locus and suggest PA3885 should be named TpbA for tyrosine phosphatase related to biofilm formation and PA1120 should be TpbB. PMID- 19543381 TI - A probabilistic model of RNA conformational space. AB - The increasing importance of non-coding RNA in biology and medicine has led to a growing interest in the problem of RNA 3-D structure prediction. As is the case for proteins, RNA 3-D structure prediction methods require two key ingredients: an accurate energy function and a conformational sampling procedure. Both are only partly solved problems. Here, we focus on the problem of conformational sampling. The current state of the art solution is based on fragment assembly methods, which construct plausible conformations by stringing together short fragments obtained from experimental structures. However, the discrete nature of the fragments necessitates the use of carefully tuned, unphysical energy functions, and their non-probabilistic nature impairs unbiased sampling. We offer a solution to the sampling problem that removes these important limitations: a probabilistic model of RNA structure that allows efficient sampling of RNA conformations in continuous space, and with associated probabilities. We show that the model captures several key features of RNA structure, such as its rotameric nature and the distribution of the helix lengths. Furthermore, the model readily generates native-like 3-D conformations for 9 out of 10 test structures, solely using coarse-grained base-pairing information. In conclusion, the method provides a theoretical and practical solution for a major bottleneck on the way to routine prediction and simulation of RNA structure and dynamics in atomic detail. PMID- 19543380 TI - Innate immune sensing of modified vaccinia virus Ankara (MVA) is mediated by TLR2 TLR6, MDA-5 and the NALP3 inflammasome. AB - Modified vaccinia virus Ankara (MVA) is an attenuated double-stranded DNA poxvirus currently developed as a vaccine vector against HIV/AIDS. Profiling of the innate immune responses induced by MVA is essential for the design of vaccine vectors and for anticipating potential adverse interactions between naturally acquired and vaccine-induced immune responses. Here we report on innate immune sensing of MVA and cytokine responses in human THP-1 cells, primary human macrophages and mouse bone marrow-derived macrophages (BMDMs). The innate immune responses elicited by MVA in human macrophages were characterized by a robust chemokine production and a fairly weak pro-inflammatory cytokine response. Analyses of the cytokine production profile of macrophages isolated from knockout mice deficient in Toll-like receptors (TLRs) or in the adapter molecules MyD88 and TRIF revealed a critical role for TLR2, TLR6 and MyD88 in the production of IFNbeta-independent chemokines. MVA induced a marked up-regulation of the expression of RIG-I like receptors (RLR) and the IPS-1 adapter (also known as Cardif, MAVS or VISA). Reduced expression of RIG-I, MDA-5 and IPS-1 by shRNAs indicated that sensing of MVA by RLR and production of IFNbeta and IFNbeta dependent chemokines was controlled by the MDA-5 and IPS-1 pathway in the macrophage. Crosstalk between TLR2-MyD88 and the NALP3 inflammasome was essential for expression and processing of IL-1beta. Transcription of the Il1b gene was markedly impaired in TLR2(-/-) and MyD88(-/-) BMDM, whereas mature and secreted IL-1beta was massively reduced in NALP3(-/-) BMDMs or in human THP-1 macrophages with reduced expression of NALP3, ASC or caspase-1 by shRNAs. Innate immune sensing of MVA and production of chemokines, IFNbeta and IL-1beta by macrophages is mediated by the TLR2-TLR6-MyD88, MDA-5-IPS-1 and NALP3 inflammasome pathways. Delineation of the host response induced by MVA is critical for improving our understanding of poxvirus antiviral escape mechanisms and for designing new MVA vaccine vectors with improved immunogenicity. PMID- 19543382 TI - Taking the lag out of jet lag through model-based schedule design. AB - Travel across multiple time zones results in desynchronization of environmental time cues and the sleep-wake schedule from their normal phase relationships with the endogenous circadian system. Circadian misalignment can result in poor neurobehavioral performance, decreased sleep efficiency, and inappropriately timed physiological signals including gastrointestinal activity and hormone release. Frequent and repeated transmeridian travel is associated with long-term cognitive deficits, and rodents experimentally exposed to repeated schedule shifts have increased death rates. One approach to reduce the short-term circadian, sleep-wake, and performance problems is to use mathematical models of the circadian pacemaker to design countermeasures that rapidly shift the circadian pacemaker to align with the new schedule. In this paper, the use of mathematical models to design sleep-wake and countermeasure schedules for improved performance is demonstrated. We present an approach to designing interventions that combines an algorithm for optimal placement of countermeasures with a novel mode of schedule representation. With these methods, rapid circadian resynchrony and the resulting improvement in neurobehavioral performance can be quickly achieved even after moderate to large shifts in the sleep-wake schedule. The key schedule design inputs are endogenous circadian period length, desired sleep-wake schedule, length of intervention, background light level, and countermeasure strength. The new schedule representation facilitates schedule design, simulation studies, and experiment design and significantly decreases the amount of time to design an appropriate intervention. The method presented in this paper has direct implications for designing jet lag, shift-work, and non-24 hour schedules, including scheduling for extreme environments, such as in space, undersea, or in polar regions. PMID- 19543384 TI - Cytoplasmic streaming in neurospora: disperse the plug to increase the flow? PMID- 19543383 TI - Global prediction of tissue-specific gene expression and context-dependent gene networks in Caenorhabditis elegans. AB - Tissue-specific gene expression plays a fundamental role in metazoan biology and is an important aspect of many complex diseases. Nevertheless, an organism-wide map of tissue-specific expression remains elusive due to difficulty in obtaining these data experimentally. Here, we leveraged existing whole-animal Caenorhabditis elegans microarray data representing diverse conditions and developmental stages to generate accurate predictions of tissue-specific gene expression and experimentally validated these predictions. These patterns of tissue-specific expression are more accurate than existing high-throughput experimental studies for nearly all tissues; they also complement existing experiments by addressing tissue-specific expression present at particular developmental stages and in small tissues. We used these predictions to address several experimentally challenging questions, including the identification of tissue-specific transcriptional motifs and the discovery of potential miRNA regulation specific to particular tissues. We also investigate the role of tissue context in gene function through tissue-specific functional interaction networks. To our knowledge, this is the first study producing high-accuracy predictions of tissue-specific expression and interactions for a metazoan organism based on whole-animal data. PMID- 19543385 TI - Preclinical evaluation of a replication-deficient intranasal DeltaNS1 H5N1 influenza vaccine. AB - BACKGROUND: We developed a novel intranasal influenza vaccine approach that is based on the construction of replication-deficient vaccine viruses that lack the entire NS1 gene (DeltaNS1 virus). We previously showed that these viruses undergo abortive replication in the respiratory tract of animals. The local release of type I interferons and other cytokines and chemokines in the upper respiratory tract may have a "self-adjuvant effect", in turn increasing vaccine immunogenicity. As a result, DeltaNS1 viruses elicit strong B- and T- cell mediated immune responses. METHODOLOGY/PRINCIPAL FINDINGS: We applied this technology to the development of a pandemic H5N1 vaccine candidate. The vaccine virus was constructed by reverse genetics in Vero cells, as a 5:3 reassortant, encoding four proteins HA, NA, M1, and M2 of the A/Vietnam/1203/04 virus while the remaining genes were derived from IVR-116. The HA cleavage site was modified in a trypsin dependent manner, serving as the second attenuation factor in addition to the deleted NS1 gene. The vaccine candidate was able to grow in the Vero cells that were cultivated in a serum free medium to titers exceeding 8 log(10) TCID(50)/ml. The vaccine virus was replication deficient in interferon competent cells and did not lead to viral shedding in the vaccinated animals. The studies performed in three animal models confirmed the safety and immunogenicity of the vaccine. Intranasal immunization protected ferrets and mice from being infected with influenza H5 viruses of different clades. In a primate model (Macaca mulatta), one dose of vaccine delivered intranasally was sufficient for the induction of antibodies against homologous A/Vietnam/1203/04 and heterologous A/Indonesia/5/05 H5N1 strains. CONCLUSION/SIGNIFICANCE: Our findings show that intranasal immunization with the replication deficient H5N1 DeltaNS1 vaccine candidate is sufficient to induce a protective immune response against H5N1 viruses. This approach might be attractive as an alternative to conventional influenza vaccines. Clinical evaluation of DeltaNS1 pandemic and seasonal influenza vaccine candidates are currently in progress. PMID- 19543388 TI - Parallel loss of plastid introns and their maturase in the genus Cuscuta. AB - Plastid genome content and arrangement are highly conserved across most land plants and their closest relatives, streptophyte algae, with nearly all plastid introns having invaded the genome in their common ancestor at least 450 million years ago. One such intron, within the transfer RNA trnK-UUU, contains a large open reading frame that encodes a presumed intron maturase, matK. This gene is missing from the plastid genomes of two species in the parasitic plant genus Cuscuta but is found in all other published land plant and streptophyte algal plastid genomes, including that of the nonphotosynthetic angiosperm Epifagus virginiana and two other species of Cuscuta. By examining matK and plastid intron distribution in Cuscuta, we add support to the hypothesis that its normal role is in splicing seven of the eight group IIA introns in the genome. We also analyze matK nucleotide sequences from Cuscuta species and relatives that retain matK to test whether changes in selective pressure in the maturase are associated with intron deletion. Stepwise loss of most group IIA introns from the plastid genome results in substantial change in selective pressure within the hypothetical RNA binding domain of matK in both Cuscuta and Epifagus, either through evolution from a generalist to a specialist intron splicer or due to loss of a particular intron responsible for most of the constraint on the binding region. The possibility of intron-specific specialization in the X-domain is implicated by evidence of positive selection on the lineage leading to C. nitida in association with the loss of six of seven introns putatively spliced by matK. Moreover, transfer RNA gene deletion facilitated by parasitism combined with an unusually high rate of intron loss from remaining functional plastid genes created a unique circumstance on the lineage leading to Cuscuta subgenus Grammica that allowed elimination of matK in the most species-rich lineage of Cuscuta. PMID- 19543386 TI - Divergent cardiopulmonary actions of heme oxygenase enzymatic products in chronic hypoxia. AB - BACKGROUND: Hypoxia and pressure-overload induce heme oxygenase-1 (HO-1) in cardiomyocytes and vascular smooth muscle cells (VSMCs). HO-1(-/-) mice exposed to chronic hypoxia develop pulmonary arterial hypertension (PAH) with exaggerated right ventricular (RV) injury consisting of dilation, fibrosis, and mural thrombi. Our objective was to identify the HO-1 product(s) mediating RV protection from hypoxic injury in HO-1(-/-) mice. METHODOLOGY/PRINCIPAL FINDINGS: HO-1(-/-) mice were exposed to seven weeks of hypoxia and treated with inhaled CO or biliverdin injections. CO reduced right ventricular systolic pressure (RVSP) and prevented hypoxic pulmonary arteriolar remodeling in both HO-1(-/-) and control mice. Biliverdin had no significant effect on arteriolar remodeling or RVSP in either genotype. Despite this, biliverdin prevented RV failure in the hypoxic HO-1(-/-) mice (0/14 manifested RV wall fibrosis or thrombus), while CO treated HO-1(-/-) mice developed RV insults similar to untreated controls. In vitro, CO inhibited hypoxic VSMC proliferation and migration but did not prevent cardiomyocyte death from anoxia-reoxygenation (A-R). In contrast, bilirubin limited A-R-induced cardiomyocyte death but did not inhibit VSMC proliferation and migration. CONCLUSIONS/SIGNIFICANCE: CO and bilirubin have distinct protective actions in the heart and pulmonary vasculature during chronic hypoxia. Moreover, reducing pulmonary vascular resistance may not prevent RV injury in hypoxia-induced PAH; supporting RV adaptation to hypoxia and preventing RV failure must be a therapeutic goal. PMID- 19543387 TI - Endogenous TasiRNAs mediate non-cell autonomous effects on gene regulation in Arabidopsis thaliana. AB - BACKGROUND: Different classes of small RNAs (sRNAs) refine the expression of numerous genes in higher eukaryotes by directing protein partners to complementary nucleic acids, where they mediate gene silencing. Plants encode a unique class of sRNAs, called trans-acting small interfering RNAs (tasiRNAs), which post-transcriptionally regulate protein-coding transcripts, as do microRNAs (miRNAs), and both sRNA classes control development through their targets. TasiRNA biogenesis requires multiple components of the siRNA pathway and also miRNAs. But while 21mer siRNAs originating from transgenes can mediate silencing across several cell layers, miRNA action seems spatially restricted to the producing or closely surrounding cells. PRINCIPAL FINDINGS: We have previously described the isolation of a genetrap reporter line for TAS3a, the major locus producing AUXIN RESPONS FACTOR (ARF)-regulating tasiRNAs in the Arabidopsis shoot. Its activity is limited to the adaxial (upper) side of leaf primordia, thus spatially isolated from ARF-activities, which are located in the abaxial (lower) side. We show here by in situ hybridization and reporter fusions that the silencing activities of ARF-regulating tasiRNAs are indeed manifested non-cell autonomously to spatially control ARF activities. CONCLUSIONS/SIGNIFICANCE: Endogenous tasiRNAs are thus mediators of a mobile developmental signal and might provide effective gene silencing at a distance beyond the reach of most miRNAs. PMID- 19543389 TI - Cardiac arrest during gamete release in chum salmon regulated by the parasympathetic nerve system. AB - Cardiac arrest caused by startling stimuli, such as visual and vibration stimuli, has been reported in some animals and could be considered as an extraordinary case of bradycardia and defined as reversible missed heart beats. Variability of the heart rate is established as a balance between an autonomic system, namely cholinergic vagus inhibition, and excitatory adrenergic stimulation of neural and hormonal action in teleost. However, the cardiac arrest and its regulating nervous mechanism remain poorly understood. We show, by using electrocardiogram (ECG) data loggers, that cardiac arrest occurs in chum salmon (Oncorhynchus keta) at the moment of gamete release for 7.39+/-1.61 s in females and for 5.20+/-0.97 s in males. The increase in heart rate during spawning behavior relative to the background rate during the resting period suggests that cardiac arrest is a characteristic physiological phenomenon of the extraordinarily high heart rate during spawning behavior. The ECG morphological analysis showed a peaked and tall T-wave adjacent to the cardiac arrest, indicating an increase in potassium permeability in cardiac muscle cells, which would function to retard the cardiac action potential. Pharmacological studies showed that the cardiac arrest was abolished by injection of atropine, a muscarinic receptor antagonist, revealing that the cardiac arrest is a reflex response of the parasympathetic nerve system, although injection of sotalol, a beta-adrenergic antagonist, did not affect the cardiac arrest. We conclude that cardiac arrest during gamete release in spawning release in spawning chum salmon is a physiological reflex response controlled by the parasympathetic nervous system. This cardiac arrest represents a response to the gaping behavior that occurs at the moment of gamete release. PMID- 19543391 TI - Mineralization potential of polarized dental enamel. AB - BACKGROUND: Management of human teeth has moved from a surgical to a more conservative approach of inhibiting or preventing lesion progression. Increasing enamel mineralization is crucial in this regard. A potential difficulty is the preferential mineralization of the outermost portion of the enamel that can prevent overall mineralization. We describe a strategy for increasing the mineralization potential of dental enamel. METHODOLOGY/PRINCIPAL FINDINGS: Extracted human premolar teeth enamel (n = 5) were exposed to a high concentration of hydrogen peroxide with an energizing source. Samples were stored in artificial saliva at 37 degrees C for 1 wk. A desktop X-ray micro-CT system was used to evaluate the mineral density of samples. Mineral distribution was polarized between the lower and the higher mineralized portion of enamel by charged oxygen free radicals due to activation of permeated hydrogen peroxide. The kinetics of energy absorption in the deeper enamel region demonstrated improvement of preferential mineralization into the region without restricting overall mineralization of the enamel. Subsequent increasing mineralization, even in the dense mineralized outer portion of enamel, was also achieved. CONCLUSIONS/SIGNIFICANCE: This increased mineralization may promote resistance to acidic deterioration of the structure. The present study is one of the primary steps towards the development of novel application in reparative and restorative dentistry. PMID- 19543390 TI - Epigenetic silencing of host cell defense genes enhances intracellular survival of the rickettsial pathogen Anaplasma phagocytophilum. AB - Intracellular bacteria have evolved mechanisms that promote survival within hostile host environments, often resulting in functional dysregulation and disease. Using the Anaplasma phagocytophilum-infected granulocyte model, we establish a link between host chromatin modifications, defense gene transcription and intracellular bacterial infection. Infection of THP-1 cells with A. phagocytophilum led to silencing of host defense gene expression. Histone deacetylase 1 (HDAC1) expression, activity and binding to the defense gene promoters significantly increased during infection, which resulted in decreased histone H3 acetylation in infected cells. HDAC1 overexpression enhanced infection, whereas pharmacologic and siRNA HDAC1 inhibition significantly decreased bacterial load. HDAC2 does not seem to be involved, since HDAC2 silencing by siRNA had no effect on A. phagocytophilum intracellular propagation. These data indicate that HDAC up-regulation and epigenetic silencing of host cell defense genes is required for A. phagocytophilum infection. Bacterial epigenetic regulation of host cell gene transcription could be a general mechanism that enhances intracellular pathogen survival while altering cell function and promoting disease. PMID- 19543392 TI - Genotyping and phylogenetic analysis of Yersinia pestis by MLVA: insights into the worldwide expansion of Central Asia plague foci. AB - BACKGROUND: The species Yersinia pestis is commonly divided into three classical biovars, Antiqua, Medievalis, and Orientalis, belonging to subspecies pestis pathogenic for human and the (atypical) non-human pathogenic biovar Microtus (alias Pestoides) including several non-pestis subspecies. Recent progress in molecular typing methods enables large-scale investigations in the population structure of this species. It is now possible to test hypotheses about its evolution which were proposed decades ago. For instance the three classical biovars of different geographical distributions were suggested to originate from Central Asia. Most investigations so far have focused on the typical pestis subspecies representatives found outside of China, whereas the understanding of the emergence of this human pathogen requires the investigation of strains belonging to subspecies pestis from China and to the Microtus biovar. METHODOLOGY/PRINCIPAL FINDINGS: Multi-locus VNTR analysis (MLVA) with 25 loci was performed on a collection of Y. pestis isolates originating from the majority of the known foci worldwide and including typical rhamnose-negative subspecies pestis as well as rhamnose-positive subspecies pestis and biovar Microtus. More than 500 isolates from China, the Former Soviet Union (FSU), Mongolia and a number of other foci around the world were characterized and resolved into 350 different genotypes. The data revealed very close relationships existing between some isolates from widely separated foci as well as very high diversity which can conversely be observed between nearby foci. CONCLUSIONS/SIGNIFICANCE: The results obtained are in full agreement with the view that the Y. pestis subsp. pestis pathogenic for humans emerged in the Central Asia region between China, Kazakhstan, Russia and Mongolia, only three clones of which spread out of Central Asia. The relationships among the strains in China, Central Asia and the rest of the world based on the MLVA25 assay provide an unprecedented view on the expansion and microevolution of Y. pestis. PMID- 19543394 TI - Induction of antibody responses to African horse sickness virus (AHSV) in ponies after vaccination with recombinant modified vaccinia Ankara (MVA). AB - BACKGROUND: African horse sickness virus (AHSV) causes a non-contagious, infectious disease in equids, with mortality rates that can exceed 90% in susceptible horse populations. AHSV vaccines play a crucial role in the control of the disease; however, there are concerns over the use of polyvalent live attenuated vaccines particularly in areas where AHSV is not endemic. Therefore, it is important to consider alternative approaches for AHSV vaccine development. We have carried out a pilot study to investigate the ability of recombinant modified vaccinia Ankara (MVA) vaccines expressing VP2, VP7 or NS3 genes of AHSV to stimulate immune responses against AHSV antigens in the horse. METHODOLOGY/PRINCIPAL FINDINGS: VP2, VP7 and NS3 genes from AHSV-4/Madrid87 were cloned into the vaccinia transfer vector pSC11 and recombinant MVA viruses generated. Antigen expression or transcription of the AHSV genes from cells infected with the recombinant viruses was confirmed. Pairs of ponies were vaccinated with MVAVP2, MVAVP7 or MVANS3 and both MVA vector and AHSV antigen specific antibody responses were analysed. Vaccination with MVAVP2 induced a strong AHSV neutralising antibody response (VN titre up to a value of 2). MVAVP7 also induced AHSV antigen-specific responses, detected by western blotting. NS3 specific antibody responses were not detected. CONCLUSIONS: This pilot study demonstrates the immunogenicity of recombinant MVA vectored AHSV vaccines, in particular MVAVP2, and indicates that further work to investigate whether these vaccines would confer protection from lethal AHSV challenge in the horse is justifiable. PMID- 19543393 TI - Differential responses of human regulatory T cells (Treg) and effector T cells to rapamycin. AB - BACKGROUND: The immunosuppressive drug rapamycin (RAPA) promotes the expansion of CD4(+) CD25(high)Foxp3(+) regulatory T cells via mechanisms that remain unknown. Here, we studied expansion, IL-2R-gamma chain signaling, survival pathways and resistance to apoptosis in human Treg responding to RAPA. METHODOLOGY/PRINCIPAL FINDINGS: CD4(+)CD25(+) and CD4(+)CD25(neg) T cells were isolated from PBMC of normal controls (n = 21) using AutoMACS. These T cell subsets were cultured in the presence of anti-CD3/CD28 antibodies and 1000 IU/mL IL-2 for 3 to 6 weeks. RAPA (1-100 nM) was added to half of the cultures. After harvest, the cell phenotype, signaling via the PI3K/mTOR and STAT pathways, expression of survival proteins and Annexin V binding were determined and compared to values obtained with freshly-separated CD4(+)CD25(high) and CD4(+)CD25(neg) T cells. Suppressor function was tested in co-cultures with autologous CFSE-labeled CD4(+)CD25(neg) or CD8(+)CD25(neg) T-cell responders. The frequency and suppressor activity of Treg were increased after culture of CD4(+)CD25(+) T cells in the presence of 1 100 nM RAPA (p<0.001). RAPA-expanded Treg were largely CD4(+)CD25(high)Foxp3(+) cells and were resistant to apoptosis, while CD4(+)CD25(neg) T cells were sensitive. Only Treg upregulated anti-apoptotic and down-regulated pro-apoptotic proteins. Treg expressed higher levels of the PTEN protein than CD4(+)CD25(neg) cells. Activated Treg+/-RAPA preferentially phosphorylated STAT5 and STAT3 and did not utilize the PI3K/mTOR pathway. CONCLUSIONS/SIGNIFICANCE: RAPA favors Treg expansion and survival by differentially regulating signaling, proliferation and sensitivity to apoptosis of human effector T cells and Treg after TCR/IL-2 activation. PMID- 19543395 TI - Natural triterpenic diols promote apoptosis in astrocytoma cells through ROS mediated mitochondrial depolarization and JNK activation. AB - BACKGROUND: Triterpene alcohols and acids are multifunctional compounds widely distributed throughout the plant kingdom that exhibit a variety of beneficial health properties, being synthetic analogs of oleanolic acid under clinical evaluation as anti-tumoral therapeutic agents. However, the antineoplastic activity of two natural occurring triterpenoid alcohols extracted from olive oil, erythrodiol (an intermediate from oleanolic acid), and its isomer, uvaol, has barely been reported, particularly on brain cancer cells. Astrocytomas are among the most common and aggressive type of primary malignant tumors in the neurological system lacking effective treatments, and in this study, we addressed the effect of these two triterpenic diols on the human 1321N1 astrocytoma cell line. PRINCIPAL FINDINGS: Erythrodiol and uvaol effectively affected cell proliferation, as well as cell cycle phases and induced 1321N1 cell death. Both triterpenes successfully modulated the apoptotic response, promoting nuclear condensation and fragmentation. They caused retraction and rounding of cultured cells, which lost adherence from their supports, while F-actin and vimentin filaments disappeared as an organized cytoplasmic network. At molecular level, changes in the expression of surface proteins associated with adhesion or death processes were also observed. Moreover, triterpene exposure resulted in the production of reactive oxygen species (ROS) with loss of mitochondrial transmembrane potential, and correlated with the activation of c-Jun N-terminal kinases (JNK). The presence of catalase reversed the triterpenic diols-induced mitochondrial depolarization, JNK activation, and apoptotic death, indicating the critical role of ROS in the action of these compounds. CONCLUSIONS: Overall, we provide a significant insight into the anticarcinogenic action of erythrodiol and uvaol that may have a potential in prevention and treatment of brain tumors and other cancers. PMID- 19543396 TI - Linking HIV-infected TB patients to cotrimoxazole prophylaxis and antiretroviral treatment in India. AB - BACKGROUND: HIV-infected persons suffering from tuberculosis experience high mortality. No programmatic studies from India have documented the delivery of mortality-reducing interventions, such as cotrimoxazole prophylactic treatment (CPT) and antiretroviral treatment (ART). To guide TB-HIV policy in India we studied the effectiveness of delivering CPT and ART to HIV-infected persons treated for tuberculosis in three districts in Andhra Pradesh, India, and evaluated factors associated with death. METHODS AND FINDINGS: We retrospectively abstracted data for all HIV-infected tuberculosis patients diagnosed from March 2007 through August 2007 using standard treatment outcome definitions. 734 HIV infected tuberculosis patients were identified; 493 (67%) were males and 569 (80%) were between the ages of 24-44 years. 710 (97%) initiated CPT, and 351 (50%) collected >60% of their monthly cotrimoxazole pouches provided throughout TB treatment. Access to ART was documented in 380 (51%) patients. Overall 130 (17%) patients died during TB treatment. Patients receiving ART were less likely to die (adjusted hazard ratio [HR] 0.4, 95% confidence interval [CI] 0.3-0.6), while males and those with pulmonary TB were more likely to die (HR 1.7, 95% CI 1.1-2.7, and HR 1.9, 95% CI 1.1-3.2 respectively). CONCLUSIONS: Among HIV infected TB patients in India death was common despite the availability of free cotrimoxazole locally and ART from referral centres. Death was strongly associated with the absence of ART during TB treatment. To minimize death, programmes should promote high levels of ART uptake and closely monitor progress in implementation. PMID- 19543398 TI - Individual differences in personality predict how people look at faces. AB - BACKGROUND: Determining the ways in which personality traits interact with contextual determinants to shape social behavior remains an important area of empirical investigation. The specific personality trait of neuroticism has been related to characteristic negative emotionality and associated with heightened attention to negative, emotionally arousing environmental signals. However, the mechanisms by which this personality trait may shape social behavior remain largely unspecified. METHODOLOGY/PRINCIPAL FINDINGS: We employed eye tracking to investigate the relationship between characteristics of visual scanpaths in response to emotional facial expressions and individual differences in personality. We discovered that the amount of time spent looking at the eyes of fearful faces was positively related to neuroticism. CONCLUSIONS/SIGNIFICANCE: This finding is discussed in relation to previous behavioral research relating personality to selective attention for trait-congruent emotional information, neuroimaging studies relating differences in personality to amygdala reactivity to socially relevant stimuli, and genetic studies suggesting linkages between the serotonin transporter gene and neuroticism. We conclude that personality may be related to interpersonal interaction by shaping aspects of social cognition as basic as eye contact. In this way, eye gaze represents a possible behavioral link in a complex relationship between genes, brain function, and personality. PMID- 19543399 TI - Motor learning characterized by changing Levy distributions. AB - The probability distributions for changes in transverse plane fingertip speed are Levy distributed in human pole balancing. Six subjects learned to balance a pole on their index finger over three sessions while sitting and standing. The Levy or decay exponent decreased as a function of learning, showing reduced decay in the probability for large speed steps and was significantly smaller in the sitting condition. However, the probability distribution for changes in fingertip speed was truncated so that the probability for large steps was reduced in this condition. These results show a learning-induced tolerance for large speed step sizes and demonstrate that motor learning in continuous tasks may be characterized by changing distributions that reflect sensorimotor skill acquisition. PMID- 19543400 TI - Cytoplasmic continuity revisited: closure of septa of the filamentous fungus Schizophyllum commune in response to environmental conditions. AB - BACKGROUND: Mycelia of higher fungi consist of interconnected hyphae that are compartmentalized by septa. These septa contain large pores that allow streaming of cytoplasm and even organelles. The cytoplasm of such mycelia is therefore considered to be continuous. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show by laser dissection that septa of Schizophyllum commune can be closed depending on the environmental conditions. The most apical septum of growing hyphae was open when this basidiomycete was grown in minimal medium with glucose as a carbon source. In contrast, the second and the third septum were closed in more than 50% and 90% of the cases, respectively. Interestingly, only 24 and 37% of these septa were closed when hyphae were growing in the absence of glucose. Whether a septum was open or closed also depended on physical conditions of the environment or the presence of toxic agents. The first septum closed when hyphae were exposed to high temperature, to hypertonic conditions, or to the antibiotic nourseothricin. In the case of high temperature, septa opened again when the mycelium was placed back to the normal growth temperature. CONCLUSIONS/SIGNIFICANCE: Taken together, it is concluded that the septal pores of S. commune are dynamic structures that open or close depending on the environmental conditions. Our findings imply that the cytoplasm in the mycelium of a higher fungus is not continuous per se. PMID- 19543397 TI - Increased frequency and compromised function of T regulatory cells in systemic sclerosis (SSc) is related to a diminished CD69 and TGFbeta expression. AB - BACKGROUND: Regulatory T cells (Tregs) are essential in the control of tolerance. Evidence implicates Tregs in human autoimmune conditions. Here we investigated their role in systemic sclerosis (SSc). METHODS/PRINCIPAL FINDINGS: Patients were subdivided as having limited cutaneous SSc (lcSSc, n = 20) or diffuse cutaneous SSc (dcSSc, n = 48). Further subdivision was made between early dcSSc (n = 24) and late dcSSc (n = 24) based upon the duration of disease. 26 controls were studied for comparison. CD3+ cells were isolated using FACS and subsequently studied for the expression of CD4, CD8, CD25, FoxP3, CD127, CD62L, GITR, CD69 using flow cytometry. T cell suppression assays were performed using sorted CD4CD25(high)CD127(-) and CD4CD25(low)CD127(high) and CD3(+) cells. Suppressive function was correlated with CD69 surface expression and TGFbeta secretion/expression. The frequency of CD4(+)CD25(+) and CD25(high)FoxP3(high)CD127(neg) T cells was highly increased in all SSc subgroups. Although the expression of CD25 and GITR was comparable between groups, expression of CD62L and CD69 was dramatically lower in SSc patients, which correlated with a diminished suppressive function. Co-incubation of Tregs from healthy donors with plasma from SSc patients fully abrogated suppressive activity. Activation of Tregs from healthy donors or SSc patients with PHA significantly up regulated CD69 expression that could be inhibited by SSc plasma. CONCLUSIONS/SIGNIFICANCE: These results indicate that soluble factors in SSc plasma inhibit Treg function specifically that is associated with altered Treg CD69 and TGFbeta expression. These data suggest that a defective Treg function may underlie the immune dysfunction in systemic sclerosis. PMID- 19543401 TI - Toll-like receptor polymorphisms and susceptibility to urinary tract infections in adult women. AB - BACKGROUND: Although behavioral risk factors are strongly associated with urinary tract infection (UTI) risk, the role of genetics in acquiring this disease is poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: To test the hypothesis that polymorphisms in Toll-like receptor (TLR) pathway genes are associated with susceptibility to UTIs, we conducted a population-based case-control study of women ages 18-49 years. We examined DNA variants in 9 TLR pathway genes in 431 recurrent cystitis (rUTI) cases, 400 pyelonephritis cases, and 430 controls with no history of UTIs. In the Caucasian subgroup of 987 women, polymorphism TLR4_A896G was associated with protection from rUTI, but not pyelonephritis, with an odds ratio (OR) of 0.54 and a 95% confidence interval (CI) of 0.31 to 0.96. Polymorphism TLR5_C1174T, which encodes a variant that abrogates flagellin induced signaling, was associated with an increased risk of rUTI (OR(95%CI): 1.81 (1.00-3.08)), but not pyelonephritis. Polymorphism TLR1_G1805T was associated with protection from pyelonephritis (OR(95%CI): 0.53 (0.29-0.96)). CONCLUSIONS: These results provide the first evidence of associations of TLR5 and TLR1 variants with altered risks of acquiring rUTI and pyelonephritis, respectively. Although these data suggest that TLR polymorphisms are associated with adult susceptibility to UTIs, the statistical significance was modest and will require further study including validation with independent cohorts. PMID- 19543402 TI - Antigen-presenting cells represent targets for R5 HIV-1 infection in the first trimester pregnancy uterine mucosa. AB - BACKGROUND: During the first trimester of pregnancy, HIV-1 mother-to-child transmission is relatively rare despite the permissivity of placental cells to cell-to-cell HIV-1 infection. The placenta interacts directly with maternal uterine cells (decidual cells) but the physiological role of the decidua in the control of HIV-1 transmission and whether decidua could be a source of infected cells is unknown. METHODOLOGY/PRINCIPAL FINDINGS: To answer to this question, decidual mononuclear cells were exposed to HIV-1 in vitro. Decidual cells were shown to be more susceptible to infection by an R5 HIV-1, as compared to an X4 HIV-1. Infected cells were identified by flow cytometry analysis. The results showed that CD14(+) cells were the main targets of HIV-1 infection in the decidua. These infected CD14(+) cells expressed DC-SIGN, CD11b, CD11c, the Fc gamma receptor CD16, CD32 and CD64, classical MHC class-I and class-II and maturation and activation molecules CD83, CD80 and CD86. The permissivity of decidual tissue was also evaluated by histoculture. Decidual tissue was not infected by X4 HIV-1 but was permissive to R5 HIV-1. Different profiles of infection were observed depending on tissue localization. CONCLUSIONS/SIGNIFICANCE: The presence of HIV-1 target cells in the decidua in vitro and the low rate of in utero mother-to-child transmission during the first trimester of pregnancy suggest that a natural control occurs in vivo limiting cell-to-cell infection of the placenta and consequently infection of the fetus. PMID- 19543403 TI - Mapping insertions, deletions and SNPs on Venter's chromosomes. AB - BACKGROUND: The very recent availability of fully sequenced individual human genomes is a major revolution in biology which is certainly going to provide new insights into genetic diseases and genomic rearrangements. RESULTS: We mapped the insertions, deletions and SNPs (single nucleotide polymorphisms) that are present in Craig Venter's genome, more precisely on chromosomes 17 to 22, and compared them with the human reference genome hg17. Our results show that insertions and deletions are almost absent in L1 and generally scarce in L2 isochore families (GC-poor L1+L2 isochores represent slightly over half of the human genome), whereas they increase in GC-rich isochores, largely paralleling the densities of genes, retroviral integrations and Alu sequences. The distributions of insertions/deletions are in striking contrast with those of SNPs which exhibit almost the same density across all isochore families with, however, a trend for lower concentrations in gene-rich regions. CONCLUSIONS: Our study strongly suggests that the distribution of insertions/deletions is due to the structure of chromatin which is mostly open in gene-rich, GC-rich isochores, and largely closed in gene-poor, GC-poor isochores. The different distributions of insertions/deletions and SNPs are clearly related to the two different responsible mechanisms, namely recombination and point mutations. PMID- 19543404 TI - Streptococcus suis, an important cause of adult bacterial meningitis in northern Vietnam. AB - BACKGROUND: Streptococcus suis can cause severe systemic infection in adults exposed to infected pigs or after consumption of undercooked pig products. S. suis is often misdiagnosed, due to lack of awareness and improper testing. Here we report the first fifty cases diagnosed with S. suis infection in northern Viet Nam. METHODOLOGY/PRINCIPAL FINDINGS: In 2007, diagnostics for S. suis were set up at a national hospital in Hanoi. That year there were 43 S. suis positive cerebrospinal fluid samples, of which S. suis could be cultured in 32 cases and 11 cases were only positive by PCR. Seven patients were blood culture positive for S. suis but CSF culture and PCR negative; making a total of 50 patients with laboratory confirmed S. suis infection in 2007. The number of S. suis cases peaked during the warmer months. CONCLUSIONS/SIGNIFICANCE: S. suis was commonly diagnosed as a cause of bacterial meningitis in adults in northern Viet Nam. In countries where there is intense and widespread exposure of humans to pigs, S. suis can be an important human pathogen. PMID- 19543405 TI - Cholinergic pairing with visual activation results in long-term enhancement of visual evoked potentials. AB - Acetylcholine (ACh) contributes to learning processes by modulating cortical plasticity in terms of intensity of neuronal activity and selectivity properties of cortical neurons. However, it is not known if ACh induces long term effects within the primary visual cortex (V1) that could sustain visual learning mechanisms. In the present study we analyzed visual evoked potentials (VEPs) in V1 of rats during a 4-8 h period after coupling visual stimulation to an intracortical injection of ACh analog carbachol or stimulation of basal forebrain. To clarify the action of ACh on VEP activity in V1, we individually pre-injected muscarinic (scopolamine), nicotinic (mecamylamine), alpha7 (methyllycaconitine), and NMDA (CPP) receptor antagonists before carbachol infusion. Stimulation of the cholinergic system paired with visual stimulation significantly increased VEP amplitude (56%) during a 6 h period. Pre-treatment with scopolamine, mecamylamine and CPP completely abolished this long-term enhancement, while alpha7 inhibition induced an instant increase of VEP amplitude. This suggests a role of ACh in facilitating visual stimuli responsiveness through mechanisms comparable to LTP which involve nicotinic and muscarinic receptors with an interaction of NMDA transmission in the visual cortex. PMID- 19543406 TI - The potential impact on farmer health of enhanced export horticultural trade between the U.K. and Uganda. AB - The export of vegetables from African countries to European markets presents consumers with an ethical dilemma: should they support local, but relatively well off farmers, or poorer farmers from distant countries? This paper considers the issue of farm worker health in the U.K. and Uganda, and considers the dilemma facing U.K. consumers if Uganda achieves their aim of exporting more vegetables to the U.K. Self-reported health scores of 1,200 farm workers in the U.K. and Uganda were measured with the internationally recognised SF-36 questionnaire and compared to an international population norm. The age-corrected health status of U.K. farm workers was significantly lower than the population norm, whereas Ugandans scored significantly higher (indicating good health) for physical health and lower for mental health. If Ugandan produce enters U.K. markets, then consumers may wish to consider both the potential benefits that enhanced trade could offer Ugandan farmers compared with its impacts on U.K. workers. PMID- 19543408 TI - Development of the Jackson Heart Study Coordinating Center. AB - The public health burden caused by cardiovascular disease (CVD) continues to adversely affect individuals in terms of cost, life expectancy, medical, pharmaceutical and hospital care. This burden has been excessive in the case of African Americans. The objective of this paper is to chronicle the procedures and processes that were implemented in the development of the Jackson Heart Study Coordinating Center. The Jackson Heart Study (JHS) is a population-based investigation of traditional and emerging risk factors that predict progression to CVD among African Americans. In response to the struggle against CVD, the Jackson Heart Study has convened a professional, technical, and administrative staff with specific competence in the operation of a coordinating center to handle the wide variety of areas related to CVD studies. The Jackson Heart Study Coordinating Center (JHSCC) was created to assure validity of the JHS findings and provide the resources necessary to meet comprehensive statistical needs (planning, implementing and monitoring data analysis); data management (designing, implementing and managing data collection and quality control), and administrative support. The JHSCC began with a commitment to support study functions in order to increase participant recruitment, retention and safety, meet regulatory requirements, prepare progress reports, and facilitate effective communication with the community and between all JHS centers. The JHSCC facilitates the efforts of the JHS scientists through the development and implementation of the study protocol. The efforts of the JHSCC have resulted in the successful preparation of scientific reports and manuscripts for publication and presentation of study findings and results. In summary, the JHSCC has emerged as an effective research mechanism that serves as the driving force behind the Jackson Heart Study activities. PMID- 19543407 TI - A review of the urban development and transport impacts on public health with particular reference to Australia: trans-disciplinary research teams and some research gaps. AB - Urbanization and transport have a direct effect on public health. A transdisciplinary approach is proposed and illustrated to tackle the general problem of these environmental stressors and public health. Processes driving urban development and environmental stressors are identified. Urbanization, transport and public health literature is reviewed and environmental stressors are classified into their impacts and which group is affected, the geographical scale and potential inventions. Climate change and health impacts are identified as a research theme. From an Australian perspective, further areas for research are identified. PMID- 19543409 TI - A systematic review of arsenic exposure and its social and mental health effects with special reference to Bangladesh. AB - Underground water in many regions of the world is contaminated with high concentrations of arsenic and the resulting toxicity has created a major environmental and public health problem in the affected regions. Chronic arsenic exposure can cause many diseases, including various physical and psychological harms. Although the physical problems caused by arsenic toxicity are well reported in literature, unfortunately the consequences of arsenic exposure on mental health are not adequately studied. Therefore we conducted a review of the available literature focusing on the social consequences and detrimental effects of arsenic toxicity on mental health. Chronic arsenic exposures have serious implications for its victims (i.e. arsenicosis patients) and their families including social instability, social discrimination, refusal of victims by community and families, and marriage-related problems. Some studies conducted in arsenic affected areas revealed that arsenic exposures are associated with various neurologic problems. Chronic arsenic exposure can lead to mental retardation and developmental disabilities such as physical, cognitive, psychological, sensory and speech impairments. As health is defined by the World Health Organization as "a state of complete physical, mental and social wellbeing", the social dimensions have a large impact on individual's mental health. Furthermore studies in China und Bangladesh have shown that mental health problems (e.g. depression) are more common among the people affected by arsenic contamination. Our study indicates various neurological, mental and social consequences among arsenic affected victims. Further studies are recommended in arsenic-affected areas to understand the underlying mechanisms of poor mental health caused by arsenic exposure. PMID- 19543410 TI - Sexual orientation, drug use preference during sex, and HIV risk practices and preferences among men who specifically seek unprotected sex partners via the internet. AB - The present study entailed conducting a content analysis of 1,434 ads/profiles posted on one of the most popular "Men who have Sex with Men" (MSM) websites that specifically fosters unprotected sex. Ads/profiles were selected randomly based on the American ZIP code of residence (n = 1,316), with a randomly-drawn oversampling of profiles of men who self-identified as heterosexual or "curious" rather than gay or bisexual (n = 118). Data were collected between September 2006 and September 2007. The purpose of the present paper is to examine the conjoint effects of self-identified sexual orientation and preference for having/not having sex while high, on men's sought-after sexual risk. Analytical comparisons of the four groups showed that, on most measures, the combination of sexual orientation and drug use preference during sex differentiated the men. Generally speaking, gay/bisexual men who advertised online for partners with whom they could have sex while high expressed the greatest interest in risky sexual behaviors (e.g., felching, unprotected oral sex, unprotected anal sex) and various risk-related preferences (e.g., multiple partner sex, anonymous sex, eroticizing ejaculatory fluids). This is especially true when they are compared to their heterosexual/"curious" counterparts whose online profiles were not as likely to indicate a desire for having sex while high. PMID- 19543411 TI - Who is exposed to secondhand smoke? Self-reported and serum cotinine measured exposure in the U.S., 1999-2006. AB - This study presents self-reported and serum cotinine measures of exposure to secondhand smoke (SHS) for nonsmoking children, adolescents, and adults. Estimates are disaggregated by time periods and sociodemographic characteristics based on analyses of the 1999-2006 National Health and Nutrition Examination Survey. Self-reported exposure rates are found to be highest for children, followed by adolescents and adults. Important differences in exposure are found by socioeconomic characteristics. Using serum cotinine to measure exposure yields much higher prevalence rates than self-reports. Rates of SHS exposure remain high, but cotinine levels are declining for most groups. PMID- 19543412 TI - The risks of inappropriateness in cardiac imaging. AB - The immense clinical and scientific benefits of cardiovascular imaging are well established, but are also true that 30 to 50% of all examinations are partially or totally inappropriate. Marketing messages, high patient demand and defensive medicine, lead to the vicious circle of the so-called Ulysses syndrome. Mr. Ulysses, a typical middle-aged "worried-well" asymptomatic subject with an A-type coronary personality, a heavy (opium) smoker, leading a stressful life, would be advised to have a cardiological check-up after 10 years of war. After a long journey across imaging laboratories, he will have stress echo, myocardial perfusion scintigraphy, PET-CT, 64-slice CT, and adenosine-MRI performed, with a cumulative cost of >100 times a simple exercise-electrocardiography test and a cumulative radiation dose of >4,000 chest x-rays, with a cancer risk of 1 in 100. Ulysses is tired of useless examinations, exorbitant costs. unaffordable even by the richest society, and unacceptable risks. PMID- 19543413 TI - Estimated time for occurrence of smoking-related consequences among pregnant and non-pregnant women. AB - OBJECTIVES: To study time estimates by women smokers for when smoking-related consequences will occur given continuing or quitting smoking. The relationship of these estimates to pregnancy and intent to quit smoking was also investigated. METHODS: Over a two-week period, eighty women, selected to constitute four subgroups formed by pregnant vs. non-pregnant and trying vs. not trying to quit smoking, rated times at which they would expect smoking-related consequences to occur given continuing or quitting smoking. RESULTS: Somatic health consequences were estimated to occur later than consequences related to mood and social relations. All consequences were estimated to occur later given quitting smoking. Pregnancy had an effect on the estimated time that consequences would occur, with pregnant women estimating earlier occurrence of consequences related to mood and social relations than non-pregnant women did. CONCLUSION: Health messages should stress consequences for somatic health in quitting smoking, since outcomes later in time might have too low a value to exert a positive effect on decisions to quit smoking. PMID- 19543414 TI - Fluoride intake through consumption of tap water and bottled water in Belgium. AB - There is a tendency to align higher levels of fluoride in natural mineral water with the existing higher levels in tap water. Treatment of natural mineral waters could harm the preservation of their natural character. In this study fluoride intake through bottled and tap water consumption in the Belgian adult population was assessed, taking into account regional differences. A deterministic approach was used whereby consumption quantities of tap water and different brands of bottled water were linked with their respective fluoride concentrations. Data from the national food consumption survey (2004) were used and the Nusser methodology was applied to obtain usual intake estimates.Mean intake of fluoride through total water consumption in Flanders was 1.4+/-0.7 mg/day (97.5(th) percentile: 3.1 mg/day), while in the Walloon region it was on average 0.9+/-0.6 mg/day (97.5(th) percentile: 2.4 mg/day). The probability of exceeding the UL of 7 mg per day via a normal diet was estimated to be low. Consequently, there is no need to revise the existing norms, but higher fluoride concentrations should be more clearly indicated on the labels. Reliable data about total dietary fluoride intake in children, including intake of fluoride via tooth paste and food supplements, are needed. PMID- 19543416 TI - Trends in Free Time with a Partner: A Transformation of Intimacy? PMID- 19543415 TI - Cigarettes butts and the case for an environmental policy on hazardous cigarette waste. AB - Discarded cigarette butts are a form of non-biodegradable litter. Carried as runoff from streets to drains, to rivers, and ultimately to the ocean and its beaches, cigarette filters are the single most collected item in international beach cleanups each year. They are an environmental blight on streets, sidewalks, and other open areas. Rather than being a protective health device, cigarette filters are primarily a marketing tool to help sell 'safe' cigarettes. They are perceived by much of the public (especially current smokers) to reduce the health risks of smoking through technology. Filters have reduced the machine-measured yield of tar and nicotine from burning cigarettes, but there is controversy as to whether this has correspondingly reduced the disease burden of smoking to the population. Filters actually may serve to sustain smoking by making it seem less urgent for smokers to quit and easier for children to initiate smoking because of reduced irritation from early experimentation. Several options are available to reduce the environmental impact of cigarette butt waste, including developing biodegradable filters, increasing fines and penalties for littering butts, monetary deposits on filters, increasing availability of butt receptacles, and expanded public education. It may even be possible to ban the sale of filtered cigarettes altogether on the basis of their adverse environmental impact. This option may be attractive in coastal regions where beaches accumulate butt waste and where smoking indoors is increasingly prohibited. Additional research is needed on the various policy options, including behavioral research on the impact of banning the sale of filtered cigarettes altogether. PMID- 19543417 TI - Novel influenza A (H1N1): where are we? PMID- 19543418 TI - Overview of asbestos issues in Korea. AB - Asbestos is a carcinogen that causes diseases such as mesothelioma and lung cancer in humans. There was a sharp increase in the use of asbestos in Korea in the 1970s as Korea's economy developed rapidly, and asbestos was only recently banned from use. Despite the ban of its use, previously applied asbestos still causes many problems. A series of asbestos-related events that recently occurred in Korea have caused the general public to become concerned about asbestos. Therefore, it is necessary to take proper action to deal with asbestos-related events, such as mass outbreaks of mesothelioma among residents who lived near asbestos textile factories or asbestos mines. Although there have been no rapid increases in asbestos-related illnesses in Korea to date, such illnesses are expected to increase greatly due to the amount of asbestos used and long latency period. Decreasing the asbestos exposure level to levels as low as possible is the most important step in preventing asbestos-related illnesses in the next few decades. However, there is a lack of specialized facilities for the analysis of asbestos and experts to diagnose and treat asbestos-related illnesses in Korea; therefore, national-level concern and support are required. PMID- 19543419 TI - Therapeutic effects of topical application of ozone on acute cutaneous wound healing. AB - This study was undertaken to evaluate the therapeutic effects of topical ozonated olive oil on acute cutaneous wound healing in a guinea pig model and also to elucidate its therapeutic mechanism. After creating full-thickness skin wounds on the backs of guinea pigs by using a 6 mm punch biopsy, we examined the wound healing effect of topically applied ozonated olive oil (ozone group), as compared to the pure olive oil (oil group) and non-treatment (control group). The ozone group of guinea pig had a significantly smaller wound size and a residual wound area than the oil group, on days 5 (P<0.05) and 7 (P<0.01 and P<0.05) after wound surgery, respectively. Both hematoxylin-eosin staining and Masson-trichrome staining revealed an increased intensity of collagen fibers and a greater number of fibroblasts in the ozone group than that in the oil group on day 7. Immunohistochemical staining demonstrated upregulation of platelet derived growth factor (PDGF), transforming growth factor-beta (TGF-beta) and vascular endothelial growth factor (VEGF) expressions, but not fibroblast growth factor expression in the ozone group on day 7, as compared with the oil group. In conclusion, these results demonstrate that topical application of ozonated olive oil can accelerate acute cutaneous wound repair in a guinea pig in association with the increased expression of PDGF, TGF-beta, and VEGF. PMID- 19543420 TI - Advanced surface reconstruction technique to build detailed surface models of the liver and neighboring structures from the Visible Korean Human. AB - Unlike volume models, surface models, which are empty three-dimensional images, have small file size, so that they can be displayed, rotated, and modified in a real time. For the reason, the surface models of liver and neighboring structures can be effectively applied to virtual hepatic segmentectomy, virtual laparoscopic cholecystectomy, and so on. The purpose of this research is to present surface models of detailed structures inside and outside the liver, which promote medical simulation systems. Forty-seven chosen structures were liver structures such as portal triad, hepatic vein, and neighboring structures such as the stomach, duodenum, muscles, bones, and skin. The structures were outlined in the serially sectioned images from the Visible Korean Human to prepare segmented images. From the segmented images, serial outlines of each structure were stacked; on the popular commercial software, advanced surface reconstruction technique was applied to build surface model of the structure. A surface model of the liver was divided into eight models of hepatic segments according to distribution of the portal vein. The surface models will be distributed to encourage researchers to develop the various kinds of medical simulation of the abdomen. PMID- 19543421 TI - Ca2+-activated K+ current in freshly isolated c-Kit positive cells in guinea-pig stomach. AB - This study was designed to isolate Ca2+-activated K+ current (I(KCa)) and elucidate its physiological significance in freshly isolated interstitial cells of Cajal (ICCs) of guinea-pig stomach. Single ICC was freshly isolated by enzymatically dissociating from myenteric border of gastric antrum free of circular muscles, and conventional whole-cell voltage clamp technique including immunohistochemical techniques were employed to characterize the cells: In myenteric border of gastric antrum, ICC-MY (ICCs from myenteric border) were detected by immunohistochemical reactivity, and single ICC-MY which has many branches was immunohistochemically c-Kit positive. Under K+-rich and 0.1 mM ethylene glycol-bis (2-aminoethyl ether)-N,N,N',N'-tetraacetic acid pipette solution, ICC produced spontaneous inward current (-256 +/- 92.2 pA). When step depolarizing pulse from -80 to +80 mV was applied at holding potential (V(h)) of 80 mV, voltage-dependent outward currents were recorded with superimposed spontaneous transient outward currents (STOCs). Both STOCs and outward currents were reversibly affected by tetraethylammonium chloride (TEA) and iberiotoxin (IbTX); 2 mM TEA and 200 nM IbTX completely abolished STOCs and significantly inhibited outward K+ current over the whole potential range tested for current/voltage (I/V) relationship. In addition, TEA delayed repolarization phase of spontaneous inward current. The present results indicate the presence of I(KCa) in a single ICC, and it might be involved in regulation of repolarizing phase of spontaneous inward current in guinea-pig stomach. PMID- 19543422 TI - Severe hypocalcemia caused by absorption of sorbitol-mannitol solution during hysteroscopy. AB - Hysteroscopic procedure can be complicated by intravascular absorption of irrigating fluid. The clinical features of this complication are similar to transurethral resection of the prostate (TURP) syndrome. There have been few reports on hypocalcemia during endoscopic surgery and clinical implications of hypocalcemia in TURP syndrome have been underestimated. We report a case of TURP syndrome association with a decreased ionized calcium concentration of 0.53 mM/L after the absorption of a large amount of sorbitol-mannitol solution during hysteroscopy. The hypotension which occurred in TURP syndrome did not respond to vasopressor and inotropic agent but responded to the administration of calcium. This case was also accompanied by hyponatremia, hyperglycemia and lactic acidosis through the metabolism of sorbitol. PMID- 19543424 TI - Giant ovarian tumor presenting as an incarcerated umbilical hernia: a case report. AB - We report a rare case of a giant ovarian tumor presenting as an incarcerated umbilical hernia. A 61-yr-old woman was admitted to the hospital with severe abdominal pain, an umbilical mass, nausea and vomiting. On examination, a large, irreducible umbilical hernia was found. The woman underwent an urgent operation for a possible strangulated hernia. A large, multilocular tumor was found. The tumor was excised, and a total abdominal hysterectomy and bilateral salphingo oophorectomy were performed. The woman was discharged 6 days after her admission. This is the first report of incarcerated umbilical hernia containing a giant ovarian tumor within the sac. PMID- 19543423 TI - GnRH agonist therapy in a patient with recurrent ovarian granulosa cell tumors. AB - A 65-yr-old woman presented 17 yr status post-hysterectomy with bilateral ovarian salpingo-oophorectomy, attributable to ovarian cancer. She was admitted to our hospital, with multiple cystic liver masses and multiple large seeded masses in her abdomen and pelvic cavity. Histological examination of the pelvic masses demonstrated granulosa cell tumors. After two courses of systemic combination chemotherapy, with paclitaxel and carboplatin, the masses in the abdomen and pelvic cavity increased, and debulking surgery also failed because of peritoneal dissemination with severe adhesion. Finally, she underwent palliative radiotherapy for only the pelvic masses obstructing the urinary and GI tracts, and monthly hormonal therapy with a gonadotrophin-releasing hormone agonist; leuprorelin 3.75 mg IM. Subsequently, multiple masses beyond the range of the radiation as well as those within the radiotherapy field partially decreased. This partial response had been maintained for more than 8 months as of the last follow-up visit. Owing to its long and indolent course and the low metabolic rate of the tumors, advanced or recurrent granulosa cell tumor (GCT) requires treatment options beyond chemotherapy, surgery, and radiotherapy. Hormonal agents may provide another treatment option for advanced or recurrent GCT in those who are not candidates for surgery, chemotherapy, or radiotherapy. PMID- 19543425 TI - Pure basaloid squamous cell carcinoma of the uterine cervix: a case report. AB - Basaloid squamous cell carcinoma of the uterine cervix is an extremely rare malignancy of the female genital tract with a poorer clinical outcome than squamous cell carcinoma of the uterine cervix. We report a case of pure basaloid squamous cell carcinoma of the uterine cervix. A 70-yr-old woman with vaginal bleeding was referred to our institute. A basaloid squamous cell carcinoma of the uterine cervix, of International Federation of Gynecology and Obstetrics (FIGO) stage Ib1, was diagnosed by a loop electrosurgical excision procedure cone biopsy. A radical hysterectomy was performed, along with bilateral salpingo oophorectomy, pelvic lymph node dissection, and para-aortic lymph node sampling. Pathologic findings were consistent with a basaloid squamous cell carcinoma confined to the cervix without an extracervical tumor. No further treatment was administered and there was no clinical evidence of recurrence during the 12 months of follow-up. Follow-up for the patient is ongoing. Although basaloid squamous cell carcinoma of the uterine cervix is thought to behave aggressively, accumulation of data on these rare tumors is necessary to determine whether their behavior differs significantly from that of conventional cervical squamous cell carcinoma of similar clinical stage. These data would be useful for defining the best diagnosis and treatment for these rare tumors. PMID- 19543426 TI - Marginal analysis of panel counts through estimating functions. AB - We develop nonparametric estimation procedures for the marginal mean function of a counting process based on periodic observations, using two types of self consistent estimating equations. The first is derived from the likelihood studied in Wellner & Zhang (2000), assuming a Poisson counting process, and gives a nondecreasing estimator, which is the same as the nonparametric maximum likelihood estimator of Wellner & Zhang and thus is consistent without the Poisson assumption. Motivated by the construction of parametric generalized estimating equations, the second type is a set of data-adaptive quasi-score functions, which are likelihood estimating functions under a mixed-Poisson assumption. We evaluate the procedures via simulation, and illustrate them with the data from a bladder cancer study. PMID- 19543427 TI - The burden of prostate cancer in Canada. AB - The clinical and economic burden of prostate cancer in Canada is substantial, and is rising. Studies indicate that 1 in 7 men will develop prostate cancer during their lifetime, and another 1 in 27 will die because of it. It is estimated that 4300 Canadian men will die of prostate cancer in 2008. Age, family history, race and diet are all risks associated with the development of prostate cancer. A diagnosis of cancer carries a significant burden and like other cancers is a cause of significant anxiety and depression. Uncertainty regarding the value of screening for prostate cancer has been, and continues to be, a challenge for primary care physicians and urologists. PMID- 19543428 TI - The role of 5-alpha reductase inhibitors in prostate pathophysiology: Is there an additional advantage to inhibition of type 1 isoenzyme? AB - Normal growth and function of the prostate are contingent on the reduction of testosterone to dihydrotestosterone (DHT) by 5-alpha reductase (5-AR) enzymes types 1 and 2. It has been theorized that an overabundance of DHT may be implicated in the pathogenesis of both benign prostatic hyperplasia (BPH) and prostate cancer. Inhibitors of 5-AR such as dutasteride and finasteride may therefore have an important role in the prevention and treatment of BPH and prostate cancer. Dutasteride provides greater suppression of DHT than finasteride, thereby underlying the hypothesis that inhibition of both type 1 and type 2 would provide correspondingly greater protection than inhibition of type 2 alone. We review the potential significance of the 5-AR inhibitors in reducing the risk of prostate cancer according to the basic biology of prostate disease. PMID- 19543429 TI - Diagnosis and management of benign prostatic hyperplasia in primary care. AB - Benign prostatic hyperplasia (BPH), and its clinical manifestation as lower urinary tract symptoms (LUTS), is a major health concern for aging men. There have been significant advances in the diagnosis and treatment of BPH in recent years. There has been a renewed interest in medical therapies and less invasive surgical techniques. As a consequence, the treatment needs of men with mild to moderate LUTS without evidence of prostate cancer can now be accomplished in a primary care setting. There are differences in the way urologists and primary care physicians approach the evaluation and management of LUTS due to BPH, which is not reflected in Canadian Urological Association (CUA) and American Urological Association (AUA) guidelines. A "shared care" approach involving urologists and primary care physicians represents a reasonable and viable model for the care of men suffering from LUTS. The essence of the model centres around educating and communicating effectively with the patient on BPH. This article provides primary care physicians with an overview of the diagnostic and management strategies outlined in recent CUA and AUA guidelines so that they may be better positioned to effectively deal with this patient population. It is now apparent that we must move away from the urologist as the first-line physician, and allow primary care physicians to accept a new role in the diagnosis and management of BPH. PMID- 19543430 TI - Gleason grading controversies: what the chemoprevention trials have taught us. AB - The recent Prostate Chemoprevention Trial (PCPT), which assessed the efficacy of finasteride in reducing prostate cancer incidence, showed promising results. However, patients who developed cancer had higher Gleason scores than those on placebo. Moreover, recent evidence has shown that the biopsy Gleason scores in patients on finasteride were actually more accurate compared with patients on placebo when matched with the final, radical prostatectomy (RP) scores. This accuracy was due to a reduction in prostate volume induced by the drug, and better performance of prostate-specific antigen correlation for identifying men with high-grade cancer. Re-evaluation of the results based on the pathology of the RP specimens and longer follow-up showed a 30% reduction in cancer incidence with finasteride and no significant differences in Gleason scores compared with placebo. PMID- 19543431 TI - Internet Daters' Body Type Preferences: Race-Ethnic and Gender Differences. AB - Employing a United States sample of 5,810 Yahoo heterosexual internet dating profiles, this study finds race-ethnicity and gender influence body type preferences for dates, with men and whites significantly more likely than women and non-whites to have such preferences. White males are more likely than non white men to prefer to date thin and toned women, while African-American and Latino men are significantly more likely than white men to prefer female dates with thick or large bodies. Compatible with previous research showing non-whites have greater body satisfaction and are less influenced by mainstream media than whites, our findings suggest Latinos and African Americans negotiate dominant white idealizations of thin female bodies with their own cultures' greater acceptance of larger body types. PMID- 19543433 TI - The Senate Electoral Cycle and Bicameral Appropriations Politics. AB - We consider the consequences of the Senate electoral cycle and bicameralism for distributive politics, introducing the concept of contested credit claiming, i.e. that members of a state's House and Senate delegations must share the credit for appropriations that originate in their chamber with delegation members in the other chamber. Using data that isolates appropriations of each chamber, we test a model of the strategic incentives contested credit claiming creates. Our empirical analysis indicates that the Senate electoral cycle induces a back loading of benefits to the end of senatorial terms, but that the House blunts this tendency with countercyclical appropriations. Our analysis informs our understanding of appropriations earmarking, and points a way forward in studying the larger consequences of bicameral legislatures. PMID- 19543432 TI - [Not Available]. AB - The scale-free and small-world network models reflect the functional units of networks. However, when we investigated the network properties of a signaling pathway using these models, no significant differences were found between the original undirected graphs and the graphs in which inactive proteins were eliminated from the gene expression data. We analyzed signaling networks by focusing on those pathways that best reflected cellular function. Therefore, our analysis of pathways started from the ligands and progressed to transcription factors and cytoskeletal proteins. We employed the Python module to assess the target network. This involved comparing the original and restricted signaling cascades as a directed graph using microarray gene expression profiles of late onset Alzheimer's disease. The most commonly used method of shortest-path analysis neglects to consider the influences of alternative pathways that can affect the activation of transcription factors or cytoskeletal proteins. We therefore introduced included k-shortest paths and k-cycles in our network analysis using the Python modules, which allowed us to attain a reasonable computational time and identify k-shortest paths. This technique reflected results found in vivo and identified pathways not found when shortest path or degree analysis was applied. Our module enabled us to comprehensively analyse the characteristics of biomolecular networks and also enabled analysis of the effects of diseases considering the feedback loop and feedforward loop control structures as an alternative path. PMID- 19543434 TI - Conceptual Hierarchies in a Flat Attractor Network: Dynamics of Learning and Computations. AB - The structure of people's conceptual knowledge of concrete nouns has traditionally been viewed as hierarchical (Collins & Quillian, 1969). For example, superordinate concepts (vegetable) are assumed to reside at a higher level than basic-level concepts (carrot). A feature-based attractor network with a single layer of semantic features developed representations of both basic-level and superordinate concepts. No hierarchical structure was built into the network. In Experiment and Simulation 1, the graded structure of categories (typicality ratings) is accounted for by the flat attractor-network. Experiment and Simulation 2 show that, as with basic-level concepts, such a network predicts feature verification latencies for superordinate concepts (vegetable ). In Experiment and Simulation 3, counterintuitive results regarding the temporal dynamics of similarity in semantic priming are explained by the model. By treating both types of concepts the same in terms of representation, learning, and computations, the model provides new insights into semantic memory. PMID- 19543435 TI - Association of Hematological Parameters with Clustered Components of Metabolic Syndrome among Professional and Office Workers in Bangkok, Thailand. AB - BACKGROUND: Accumulating evidence documents associations between alterations in hematological parameters, indicative of prothrombotic and proinflammatory states, and risk of metabolic syndrome (MetS). We investigated associations of hematological parameters with MetS and individual criteria of the syndrome among Thai professional and office workers. METHODS: Study subjects were 1,314 patients (531 men and 783 women) who participated in annual health examinations during the period of August through December 2001. MetS was defined using the modified ATP III criteria. Multivariable logistic regression procedures were used to estimate odds ratios (OR) and 95% confidence intervals (95%CI) of MetS risk according to quartiles of each hematological parameter with the lowest quartile specified as the referent group. RESULTS: WBC counts increased with increasing numbers of MetS components in both men and women. Among women, platelet counts, hemoglobin and hematocrit concentrations increased with increasing numbers of MetS components (p<0.05). No similar trends were observed for men. Of the hematological parameters studied, elevated platelet and WBC were statistically significantly associated with MetS among men (OR=1.86, 95% CI: 1.03-3.36; OR=2.26, 95% CI: 1.27-4.02), respectively. Among women, MetS risk increased across successive quartiles of hemoglobin (1.00, 2.63, 3.59 and 4.36; p for trend = 0.002), hematocrit (1.00, 2.35, 3.04 and 5.70; p-for trend <0.001), platelet (1.00, 2.37, 2.83 and 3.11; p-for trend = 0.014) and WBC counts (1.00, 2.97, 4.09 and 5.41; p-for trend < 0.001). CONCLUSIONS: Our data are consistent with an emerging literature demonstrating altered hematological status in patients at high risk of MetS. PMID- 19543436 TI - A New Synthetic Method for the Preparation of Protonated-NHCs and Related Compounds. AB - Protonated versions of N-heterocyclic carbenes (NHC,H(+)) are classically prepared by closing the ring through the introduction of the CH+ fragment. He we report a totally different synthetic approach, which can be viewed as the addition of a 1,3-diazaallyl anion to a compound featuring two leaving groups (hereafter named "di-electrophile"). Using 1,3- and 1,4-dibromides, six and seven membered NHC,H(+)s have been prepared in good yields. Similarly, with 1,3,2 dioxathiolane-2,2-dioxide as a di-electrophile, imidazolidinium salts were obtained. To illustrate its broad scope of application, this synthetic route has been expanded to the preparation of protonated cyclic amino alkyl carbenes (CAACs) and amino thio carbenes, using 1-aza-allyl and 1,3-azathio-allyl anions, respectively. PMID- 19543437 TI - A Stable (Amino)(Phosphino)Carbene as bidentate ligand for Palladium and Nickel Complexes: Synthesis, Structure, and Catalytic Activity. AB - Two original complexes featuring an (amino)(phosphino)carbene eta(2)-bonded to the metal have been obtained in 60 and 80 % yields, by addition of the corresponding stable carbene to PdCl(2)(cod) and NiCl(2)(PPh(3))(2), respectively. Both complexes have been fully characterized including X-ray diffraction studies. The catalytic activity of the palladium complex has been evaluated for aryl amination reactions. PMID- 19543438 TI - Both exogenous and endogenous target salience manipulations support resource depletion accounts of the attentional blink: A reply to Olivers et al. AB - Input-control theories of the attentional blink (AB) suggest that this deficit results from impaired attentional selection caused by the post-Target 1 (T1) distractor (Di Lollo et al., 2005; Olivers et al., 2007). Accordingly, there should be no AB when there are no intervening distractors between the targets. Contrary to these hypotheses, Dux et al. (2008) observed an AB (T3 deficit) when three targets, from the same attentional set, were presented successively in a rapid stream of distractors if subjects increased the resources devoted to T1 processing, a result consistent with resource depletion accounts of the AB. However, Olivers et al. (this issue) argue that Dux et al.'s results can be better explained by the relationship between T1 and T2 rather than between T1 and T3, and by target discriminability effects. Here, we find that manipulating the resources subjects devote to T1, either exogenously (target perceptual salience) or endogenously (target task-relevance), affects T3 performance even when controlling for T2 and target discriminability differences. These results support Dux et al.'s conclusion that T1 resource depletion underlies the AB. PMID- 19543440 TI - Silicon Wafer-Based Platinum Microelectrode Array Biosensor for Near Real-Time Measurement of Glutamate in Vivo. AB - Using Micro-Electro-Mechanical-Systems (MEMS) technologies, we have developed silicon wafer-based platinum microelectrode arrays (MEAs) modified with glutamate oxidase (GluOx) for electroenzymatic detection of glutamate in vivo. These MEAs were designed to have optimal spatial resolution for in vivo recordings. Selective detection of glutamate in the presence of the electroactive interferents, dopamine and ascorbic acid, was attained by deposition of polypyrrole and Nafion. The sensors responded to glutamate with a limit of detection under 1muM and a sub-1-second response time in solution. In addition to extensive in vitro characterization, the utility of these MEA glutamate biosensors was also established in vivo. In the anesthetized rat, these MEA glutamate biosensors were used for detection of cortically-evoked glutamate release in the ventral striatum. The MEA biosensors also were applied to the detection of stress-induced glutamate release in the dorsal striatum of the freely-moving rat. PMID- 19543439 TI - Appetite for destruction: E3 ubiquitin-ligase protection in cardiac disease. AB - Over the course of 3 billion heartbeats in an average human lifetime, the heart must maintain constant protein quality control, including the coordinated and regulated degradation of proteins via the ubiquitin-proteasome system (UPS). Recent data highlight the specificity by which the UPS functions in the context of cardiac hypertrophy, ischemic heart disease and cardiomyopathies. Although curbing the appetite of the proteasome through the use of inhibitors in animal models of cardiac disease has proven effective experimentally, recent studies report proteasome inhibition as being cardiotoxic in some patients. Therefore, focusing on specific regulatory components of the proteasome, such as members of the E3 ubiquitin-ligase family of proteins, may hold promise for targeted therapeutics of cardiac disease. This review focuses on the UPS, its specific role in cardiac disease and opportunities for novel therapies. PMID- 19543441 TI - Anti-CTLA4 Antibody Clinical Trials in Melanoma. AB - The cytotoxic T lymphocyte-associated protein 4 (CTLA4) is a main negative regulator of the immune system, which inhibits the costimulatory signaling for T cells. Preclinical studies demonstrated that antibodies against CTLA4 induced regression of some murine tumors. Two CTLA4 blocking monoclonal antibodies have entered clinical development and are currently in pivotal clinical trial testing. Ipilimumab (formerly MDX010) is an IgG1 and tremelimumab (formerly CP-675,206 and transiently ticilimumab), is an IgG2, both being fully human monoclonal antibodies. Across several early clinical trials, including dose escalation, single dose, multi-dose, and in combination with a variety of other immune stimulants like peptide vaccines or interleukin-2, objective tumor responses in patients with metastatic melanoma have been observed in the in the range of 5 to 20%. A key feature is that some of these responses are extremely long-lived responses, lasting years. The early clinical testing also demonstrated that these CTLA4 blocking antibodies can lead to significant toxicities, most with an inflammatory or immune mediated mechanism of action. These include colitis and skin rash as the most common toxicities, and a variety of autoimmune and inflammatory processes against multiple organs. Some of these toxicities require immune suppressive therapy and may lead to permanent damage in occasional patients. In conclusion, two monoclonal antibodies blocking CTLA4 have demonstrated ability to break tolerance to self-tissues and result in long lasting objective cancer regressions, and have moved onto late stages of clinical development. PMID- 19543443 TI - A Field Experiment Testing the Utility of Regulatory Fit Messages for Promoting Physical Activity. AB - Guided by regulatory focus theory, we examined whether messages tailored to individuals' promotion- or prevention-goal orientation (regulatory focus) elicit positive thoughts and feelings about physical activity and increase participation in physical activity. Inactive participants (N = 206) were assigned randomly to receive either promotion-focused or prevention-focused messages encouraging physical activity. Two weeks after message exposure, we assessed participants' thoughts and feelings about physical activity and physical activity behavior. Tailored messages that fit individuals' regulatory focus led to greater physical activity participation and more positive feelings than non-fit messages, particularly in the promotion-focused condition. Furthermore, positive retrospective feelings about physical activity mediated the effects of the tailored messages on behavior. These findings provide support for regulatory focus theory and direction for enhancing the effectiveness of messages encouraging physical activity and other health behaviors. PMID- 19543442 TI - Silk as a Biomaterial. AB - Silks are fibrous proteins with remarkable mechanical properties produced in fiber form by silkworms and spiders. Silk fibers in the form of sutures have been used for centuries. Recently regenerated silk solutions have been used to form a variety of biomaterials, such as gels, sponges and films, for medical applications. Silks can be chemically modified through amino acid side chains to alter surface properties or to immobilize cellular growth factors. Molecular engineering of silk sequences has been used to modify silks with specific features, such as cell recognition or mineralization. The degradability of silk biomaterials can be related to the mode of processing and the corresponding content of beta sheet crystallinity. Several primary cells and cell lines have been successfully grown on different silk biomaterials to demonstrate a range of biological outcomes. Silk biomaterials are biocompatible when studied in vitro and in vivo. Silk scaffolds have been successfully used in wound healing and in tissue engineering of bone, cartilage, tendon and ligament tissues. PMID- 19543444 TI - COCIS: Markov processes in single molecule fluorescence. AB - This article examines the current status of Markov processes in single molecule fluorescence. For molecular dynamics to be described by a Markov process, the Markov process must include all states involved in the dynamics and the FPT distributions out of those states must be describable by a simple exponential law. The observation of non-exponential first-passage time distributions or other evidence of non-Markovian dynamics is common in single molecule studies and offers an opportunity to expand the Markov model to include new dynamics or states that improve understanding of the system. PMID- 19543445 TI - Exploring the Roles of Extracurricular Activity Quantity and Quality in the Educational Resilience of Vulnerable Adolescents: Variable- and Pattern-Centered Approaches. AB - This longitudinal study examines how extracurricular activity involvement contributes to "educational resilience"-the unexpected educational attainments of adolescents who are otherwise vulnerable to curtailed school success due to personal- and social-level risks. Educationally vulnerable youth characterized by significant risks and an absence of assets were identified during early adolescence (approximately age 14) using measures of academic motivation, achievement, and mental health as well as family, school, and peer contexts. Using a mixture of variable- and pattern-centered analytic techniques, we investigate how both the total amount time that vulnerable youth spent in positive extracurricular activities and the specific pattern of their extracurricular activity involvement during late adolescence (approximately age 17) predict their subsequent enrollment in college during early adulthood (up through approximately age 21). Educational resilience was predicted uniquely by some, but not all, activity patterns. These results suggest that positive extracurricular activity settings afford vulnerable youth developmentally appropriate experiences that promote educational persistence and healthy development. PMID- 19543446 TI - Synthesis of linoleic acids combinatorially-labeled at the vinylic positions as substrates for lipoxygenases. AB - Mammalian lipoxygenases have been implicated in a number of inflammation-related human diseases. Soybean lipoxygenase-1 is the archetypical example of known lipoxygenases. Here we report the synthesis of linoleic acid and (11,11)-d2 linoleic acid which are combinatorially labeled at the vinylic positions (9, 10, 12, and 13). Combinatorial labeling schemes allow for the simultaneous determination of KIEs in enzymatic reactions using NMR. Substrates are, thus, available as probes of detailed mechanism in kinetic isotope effect (KIE) studies of lipoxygenases. PMID- 19543447 TI - Surface Modification of Protein Nanocontainers and Their Self-Directing Character in Polymer Blends. AB - Tailoring the surfaces of a nanocontainer with polymer brushes that have different affinities to the components of a phase-separating polymer blend should impart self-directing properties to the nanocontainers. Such nanocontainers could then be used to deliver a variety of functional species in tunable amounts and in a site-specific manner to polymer systems. This paper describes the surface modification, subsequent characterization of nanocontainers derived from ferritin, and the effects of surface modification on their self-directing properties in a binary phase separating homopolymer blend. Wild ferritin was either PEGylated or alkylated by zero-length crosslinking to its surface carboxylate groups that were activated by carbodiimide. Modification was confirmed by ion-exchange chromatography, zeta-potential measurement, and electrospray ionization mass spectrometry. FT-IR spectrometry was used to quantify the extent of PEGylation by ratioing the intensity of the C-O-C asymmetric stretching vibration from the grafted PEG to that of the carbonyl stretching vibration (amide I band) from the protein. Importantly, modified ferritin was soluble in the organic solvent dichloromethane (DCM). Modified ferritin was introduced into a polymer blend of hydrophobic and hydrophilic polymers made up of poly (desaminotyrosyl tyrosine dodecyl ester carbonate) (PDTD) and PEG by solvent casting from solution in the common solvent DCM. Polymer thin films with an average thickness of ~ 200 mum were obtained upon evaporation of the solvent. Transmission electron micrographs of microtomed polymer films demonstrated remarkable selectivity of PEGylated ferritin to PEG domains, while alkylated ferritin self-directs to the PDTD matrix. PMID- 19543448 TI - Caged O-phosphorothioyl amino acids as building blocks for Fmoc-based solid phase peptide synthesis. AB - The synthesis of 1-(2-nitrophenylethyl) caged O-phosphorothioylserine, -threonine and -tyrosine derivatives is reported. These amino acid building blocks can be directly incorporated into peptides by Fmoc-based solid phase synthesis as their pentafluorophenyl esters or as symmetric anhydrides. Upon irradiation with UV light, the thiophosphate group, representing a hydrolysis resistant phosphate analog, is revealed. PMID- 19543449 TI - [Not Available]. AB - There has been substantial recent growth in the use of non-invasive optical brain imaging in studies of human brain function in health and disease. Near-infrared neuroimaging (NIN) is one of the most promising of these techniques and, although NIN hardware continues to evolve at a rapid pace, software tools supporting optical data acquisition, image processing, statistical modeling, and visualization remain less refined. Python, a modular and computationally efficient development language, can support functional neuroimaging studies of diverse design and implementation. In particular, Python's easily readable syntax and modular architecture allow swift prototyping followed by efficient transition to stable production systems. As an introduction to our ongoing efforts to develop Python software tools for structural and functional neuroimaging, we discuss: (i) the role of non-invasive diffuse optical imaging in measuring brain function, (ii) the key computational requirements to support NIN experiments, (iii) our collection of software tools to support NIN, called NinPy, and (iv) future extensions of these tools that will allow integration of optical with other structural and functional neuroimaging data sources. Source code for the software discussed here will be made available at www.nmr.mgh.harvard.edu/Neural_SystemsGroup/software.html. PMID- 19543450 TI - [Not Available]. AB - The Parallel Circuit SIMulator (PCSIM) is a software package for simulation of neural circuits. It is primarily designed for distributed simulation of large scale networks of spiking point neurons. Although its computational core is written in C++, PCSIM's primary interface is implemented in the Python programming language, which is a powerful programming environment and allows the user to easily integrate the neural circuit simulator with data analysis and visualization tools to manage the full neural modeling life cycle. The main focus of this paper is to describe PCSIM's full integration into Python and the benefits thereof. In particular we will investigate how the automatically generated bidirectional interface and PCSIM's object-oriented modular framework enable the user to adopt a hybrid modeling approach: using and extending PCSIM's functionality either employing pure Python or C++ and thus combining the advantages of both worlds. Furthermore, we describe several supplementary PCSIM packages written in pure Python and tailored towards setting up and analyzing neural simulations. PMID- 19543451 TI - ? AB - We present a case of a 22-year-old man who presented with traumatic unilateral testicular dislocation resulting from a blunt scrotal injury. Colour flow Doppler imaging revealed a viable testis dislocated in the inguinal canal. Inguinal exploration confirmed a healthy and viable testis, which was relocated in the scrotum and an orchiopexy was performed. The patient made an uneventful recovery. We recommend early intervention once the diagnosis of testicular dislocation is established to preserve testicular function. PMID- 19543452 TI - ? AB - We report the case of a 56-year-old postmenopausal woman who presented with incidental left hydronephrosis during an investigation for a gastrointestinal complaint. The patient denied any history of flank pain or hematuria. Contrast enhanced computed tomography revealed severe right-sided ureterohydronephrosis as well as renal atrophy. The contralateral kidney was normal, as was the patient's overall renal function. A retrograde ureterogram demonstrated complete ureteral obstruction 4 cm proximal to the ureterovesical junction. Subsequent ureteroscopy revealed a polypoid mass completely occupying the ureteral lumen, of which the biopsies demonstrated inconclusive atypical urothelial changes. The patient underwent a laparoscopic nephrectomy with open dissection of the distal ureter. The patient recovered well postoperatively. Final pathology revealed a benign obstructing endometrioma without evidence of invasion from periureteral tissue. This appears to be the first reported case of asymptomatic primary ureteral endometrioma with secondary renal atrophy. Earlier investigation and treatment may have allowed for renal preservation earlier in the course of the disease. PMID- 19543453 TI - ? AB - Castleman disease (CD) is an uncommon lymphoproliferative disorder and is especially rare in the retroperitoneum or perirenal area. We report the case of a 42-year-old woman in whom we found on routine physical examination a mass localized in the right abdomen without clinically important constitutional symptoms for 30 days. Abdominal ultrasound, magnetic resonance images and computed tomography scans revealed a solid mass localized just below the right kidney in the retroperitoneum. The patient subsequently underwent an exploratory laparotomy. Pathological examination revealed a right retroperitoneal mass of CD (hyaline vascular type). The patient completely recovered after surgery. Castleman disease is commonly misdiagnosed as malignant lymphoma, lymphadenitis or ectopic thymoma. So far, its diagnosis is mainly achieved via histopathological examination of surgically obtained tissue. After removal of the lesion, local CD has a good prognosis. PMID- 19543454 TI - ? AB - A 50-year-old woman presented with microscopic hematuria. Initial radiological investigations suggested the diagnosis of renal calculi in the left kidney. However, further assessment confirmed a renal artery aneurysm. We discuss the differential diagnosis of radio-opaque densities in the renal area. PMID- 19543455 TI - ? AB - We present the case of a 70-year-old man who had a prostate adenocarcinoma that metastatized to a previously unknown cranial meningioma. Central nervous system (CNS) metastases are very uncommon in patients with prostate cancer, and metastases to pre-existing primary CNS tumours are even more uncommon. Rare events like this can cause diagnostic uncertainty, as shown by this case. This case is a reminder for clinicians to consider prostate metastases in patients with known prostate carcinoma and focal neurological symptoms. PMID- 19543456 TI - ? AB - Carcinoid tumours, most frequently reported in the gastrointestinal and respiratory tracts, are exceedingly rare primary renal cancers. Few cases have been published to date. To our knowledge, we report the first case of a primary carcinoid tumour of the kidneys involving the inferior vena cava. We treated a 58 year-old woman with an open radical nephrectomy and cavotomy with thrombectomy. We describe the presentation, investigations and pathology results. We discuss the current experience with carcinoid tumours as a literature review relating to the diagnosis of the disease and the prognosis of patients with this neoplasm. Localized carcinoid tumours of the kidneys, including those involving the vena cava, can be successfully treated with surgical excision. PMID- 19543457 TI - [Not Available]. PMID- 19543458 TI - [Not Available]. PMID- 19543459 TI - ? PMID- 19543460 TI - ? AB - Difficulty in Foley catheter placement is a frequently encountered problem. We describe a simple and safe technique for this condition. Rather than using force, which may lead to the formation of a false passage, one should place a glidewire into the bladder through the area of resistance, followed by the placement of a Foley catheter over the glidewire. This is a very easy procedure and can be taught to nurses and nurse practitioners to avoid an unnecessary call for a urologist in the emergency department. PMID- 19543461 TI - ? AB - INTRODUCTION: Canadian health care policy faces unprecedented pressures to reform. With new advances in health care technologies and treatments, proven difficulties in obtaining timely access to necessary health care and the realities of limited fiscal resources sinking in, the status quo is being challenged with the increased role of privately funded health care. To assess the opinions of Canadian urologists on privatization of health care in Canada, the Socioeconomic Committee of the Canadian Urological Association (CUA) surveyed all active members on their beliefs on the role and impact private health care should have in urology. METHODS: We emailed a short survey of 9 questions proposed by the CUA Socioeconomic Committee regarding private health care delivery to all active CUA members in April 2007. We received responses by email, fax or mail over a 1-month period. RESULTS: Of the 440 emails sent out, 90 surveys were returned. Respondents believed that a parallel private heath care system would shorten wait times and improve access to care (74%), improve outcomes for those with private health care (58.8%), would not impair the outcomes of those without private health care (74.2%) and would not interfere with the accessibility of health care for most Canadians (73.3%). Most respondents (91.1%) believed that, if privately delivered health care was allowed, urologists should spend a fixed amount of time providing services within the public health care system as well. CONCLUSION: This survey on Canadian urologists' beliefs on the role and impact private health care should have in urology indicated that most respondents anticipate a growing influence of private health care and advocate for a regulated fixed proportion of service dedicated to the public system. PMID- 19543462 TI - ? PMID- 19543463 TI - ? AB - INTRODUCTION: Prostate cancer is the most common nonskin malignancy affecting men and is the second leading cause of cancer-related death in North America. The incidence of prostate cancer increases dramatically with age. However, many health authorities advocate the cessation of routine prostate cancer testing in men older than 75 because of the belief that most patients will have a clinically insignificant cancer and will not benefit from therapy. The true prevalence of clinically significant prostate cancer in elderly men is not known. METHODS: We analyzed 1446 needle biopsies of the prostate in men aged 75 or older. All pathological reviews were conducted by the pathology department at the Methodist Hospital in Houston, Tex. Data were collected from pathology reports, hospital and clinic databases, and medical records when available. Data obtained included age at biopsy, serum prostate-specific antigen (PSA) levels, number of positive core biopsies and Gleason grade. Statistical analysis was performed using Stata. Clinically significant cancer was defined by the pathological presence of Gleason grade 6 adenocarcinoma in more than 1 biopsy core or the presence of any Gleason 4 or 5 component in the biopsy. RESULTS: The median age of the patients included in the study was 78.8 and 95% of the patients were between the ages of 75 and 85. The mean serum PSA level for patients biopsied was 10.4 mug/L. Of all biopsies reviewed, 53% were positive for prostate cancer and 78% of these would be defined as clinically significant cancer. Regression analysis revealed age to be a significant (p < 0.05) factor for increased Gleason grade in positive biopsies. Logistic regression revealed age as a significant factor (p < 0.05) for clinically significant prostate cancer even when controlling for PSA. A serum PSA threshold value of 6.5 mug/L would have missed 38% of significant cancers and a threshold of 4.0 mug/L would have missed 8% of significant cancers. CONCLUSION: Our findings suggest that the prevalence of clinically significant prostate cancer in the elderly population may be higher than previously thought. As the population continues to live longer and healthier lives, it will become more common to confront prostate cancer morbidity in the eldery population. Using higher serum PSA thresholds to eliminate unnecessary biopsies in older men does not appear to help identify patients at greater risk of having clinically significant prostate cancer. Patients with prostate cancer having aggressive clinical features may benefit from treatment of their prostate cancer well into their eighth and ninth decades of life. Testing and diagnostic recommendations should reflect the potential benefit of identifying patients with aggressive prostate cancer even after age 75. PMID- 19543464 TI - ? PMID- 19543465 TI - Early detection of prostate cancer local recurrence by urinary prostate-specific antigen. AB - PURPOSE: We assessed the role of urinary prostate-specific antigen (uPSA) in the follow-up of prostate cancer after retropubic radical prostatectomy (RRP) for the early detection of local recurrences. METHODS: We recruited 50 patients previously treated for prostate cancer with RRP and who had not experienced a prostate-specific antigen (PSA) recurrence within their first postoperative year into a cross-sectional laboratory assessment and prospective 6-year longitudinal follow-up study. We defined biochemical failure as a serum PSA (sPSA) of 0.3 MUg/L or greater. Patients provided blood samples and a 50-mL sample of first voided urine. We performed Wilcoxon rank-sum and Fisher exact tests for statistical analysis. RESULTS: The median sPSA was 0.13 MUg/L. The median uPSA was 0.8 MUg/L, and was not significantly different when comparing Gleason scores or pathological stages. Of the 50 patients, 27 initially had a nondetectable sPSA but a detectable uPSA, and 11 patients experienced sPSA failure after 6 years. Six patients had detectable sPSA and uPSA initially. Fifteen patients were negative for both sPSA and uPSA, and 13 remained sPSA-free after 6 years. The odds ratio (OR) of having sPSA failure given a positive uPSA test was 4.5 if sPSA was undetectable, but was reduced to 2.6 if sPSA was detectable. The pooled Mantel-Haenszel OR of 4.2 suggested that a detectable uPSA quadrupled the risk of recurrence, independent of whether sPSA was elevated or not. The sensitivity of uPSA for detecting future sPSA recurrences was 81% and specificity was 45%. CONCLUSION: Urinary PSA could contribute to an early detection of local recurrences of prostate cancer after a radical prostatectomy. PMID- 19543466 TI - ? PMID- 19543467 TI - ? AB - BACKGROUND: We performed a retrospective chart review in 2006 to review oncology patients' use of banked semen samples in fertility treatments at a tertiary care centre. METHODS: From 2002 to 2005, 367 oncology patients banked semen. During the same period, 31 patients used banked samples in 48 treatment cycles. Samples were used for intrauterine insemination (IUI) in 28 cycles and for in vitro fertilization (IVF) with or without intracytoplasmic sperm injection (ICSI) in 20 cycles. RESULTS: Pregnancy rates per cycle were 21% for IUI and 50% for IVF with or without ICSI. Overall, 16 of the 31 couples achieved a pregnancy with assisted reproductive technologies (52%). CONCLUSION: This data indicates high pregnancy success rates with the use of banked semen samples from men with cancer. PMID- 19543468 TI - ? PMID- 19543469 TI - ? AB - INTRODUCTION: The endoscopic management of vesicoureteral reflux (VUR) with subureteric injection (STING) has become more popular. The low morbidity associated with the STING procedure has led to some authors advocating its use as a first-line therapy. Many parents are uncomfortable with this procedure being performed in children because of the potential morbidity associated with general anesthesia. We present an alternative without added anesthetic morbidity: offering the parents a STING when their child is undergoing an anesthetic for another surgical indication. METHODS: We reviewed the records of 10 children who underwent incidental dextranomer/hyaluronic acid copolymer (DHA) injection over a 2-year period. RESULTS: We considered the treatment outcome after a single STING procedure to be successful in 8 (80%) patients and a failure in 2 (20%). Distribution of VUR grade, according to the highest grade per patient, was high in 5 (50%) patients, moderate in 3 (30%) and low in 2 (20%). We observed no complications. CONCLUSION: The idea of performing STING in children under incidental anesthetic introduces yet another possibility in the paradigm of VUR care. Though the long-term efficacy of DHA remains to be determined, this option reduces the potential morbidity of DHA as first-line therapy while favourably altering the cost benefit. PMID- 19543470 TI - ? PMID- 19543472 TI - ? PMID- 19543471 TI - ? AB - The development, advancement and clinical integration of robotic technology in surgery continue at a staggering pace. In no other discipline has this rapid evolution occurred to a greater degree than in urology. Although radical prostatectomy has grown to become the prototypical application for the robot, the role of the robot in renal surgery remains controversial. Herein we review the literature on robotic renal surgery. A comprehensive PubMed literature search was performed to identify all published reports relating to robotic renal surgery. All clinically related articles involving human participants were critically appraised in this review. Fifty-one clinical articles were included, encompassing robot-assisted pyeloplasty, nephrectomy, nephroureterectomy, living-donor nephrectomy and partial nephrectomy. Feasibility has been shown for each of these procedures. Robot-assisted techniques have been described for almost all renal related procedures. However, the intersect between feasibility and necessity as it pertains to robotic renal surgery has yet to be defined. Also, the high cost of surgical robotic technology mandates critical appraisal before adoption, especially in a publicly funded health care system, such as the one present in Canada. PMID- 19543474 TI - ? PMID- 19543473 TI - ? PMID- 19543475 TI - ? PMID- 19543476 TI - ? PMID- 19543477 TI - ? PMID- 19543478 TI - ? PMID- 19543479 TI - Radiation-stimulated epigenetic reprogramming of adaptive-response genes in the lung: an evolutionary gift for mounting adaptive protection against lung cancer. AB - Humans are continuously exposed to low-level ionizing radiation from natural sources. However, harsher radiation environments persisted during our planet's early years and mammals survived via an evolutionary gift--a system of radiation induced natural protective measures (adaptive protection). This system includes antioxidants, DNA repair, apoptosis of severely damaged cells, epigenetically regulated apoptosis (epiapoptosis) pathways that selectively remove precancerous and other aberrant cells, and immunity against cancer. We propose a novel model in which the protective system is regulated at least in part via radiation-stress stimulated epigenetic reprogramming (epireprogramming) of adaptive-response genes. High-dose radiation can promote epigenetically silencing of adaptive response genes (episilencing), for example via promoter-associated DNA and/or histone methylation and/or histone deacetylation. Evidence is provided for low linear-energy-transfer (LET) radiation-activated natural protection (ANP) against high-LET alpha-radiation-induced lung cancer in plutonium-239 exposed rats and radon-progeny-exposed humans. Using a revised hormetic relative risk model for cancer induction that accounts for both epigenetic activation (epiactivation) and episilencing of genes, we demonstrate that, on average, >80% of alpha-radiation induced rat lung cancers were prevented by chronic, low-rate gamma-ray ANP. Interestingly, lifetime exposure to residential radon at the Environmental Protection Agency's action level of 4 pCi L(-1) appears to be associated with on average a > 60% reduction in lung cancer cases, rather than an increase. We have used underlined italics to indicate newly introduced terminology. PMID- 19543481 TI - The stimulatory effects of topical application of radioactive lantern mantle powder on wound healing. AB - Some people in different parts of Iran use burned mantles as a wound healing medicine. To perform surface area measurement, twenty rats were divided randomly into two groups of 10 animals each. The 1st group received topical burned radioactive lantern mantle powder at 1st-3rd day after making excision wounds. The 2nd group received non-radioactive lantern mantle powder. For histological study, 36 male rats randomly divided into two groups of 18 animals each. Full thickness excision wound (314+/-31.4 mm(2)) was made on the dorsal neck in all animals after inducing general anesthesia. For the first 3 days, cases received topical application of the radioactive lantern mantle powder. Finally, to measure the tensile strength, an incision was made on the dorsal neck of the rats. Surface area measurement of the wounds showed a progressive surface reduction in both groups. Histological study showed a significant statistically difference between cases and controls with respect to fibrinoid necrosis and neutrophilic exudate at the days 3 and 14. Considering the existence of granulation tissue, a significant difference was observed between case and control groups at days 3 and 7. Tensile strength study showed no significant difference between the cases and controls until 30 days after excision. PMID- 19543482 TI - Quantification of hormesis in anticancer-agent dose-responses. AB - Quantitative features of dose responses were analyzed for 2,189 candidate anticancer agents in 13 strains of yeast (Saccharomyces cerevisiae). The agents represent a diverse class of chemical compounds including mustards, other alkylating agents, and antimetabolites, inter alia. Previous analyses have shown that the responses below the toxic threshold were stimulatory and poorly predicted by a threshold dose-response model, while better explained by a hormetic dose-response model. We determined the quantitative features of the hormetic concentration-responses (n = 4,548) using previously published entry and evaluative criteria. The quantitative features that are described are: (1) the width of the concentration range showing stimulation above 10% of the control (mean of 5-fold), (2) the maximum stimulation of the concentration-responses (mean of 27% above the control), and (3) the width from the maximum stimulation to the toxicological threshold (mean of 3.7-fold). These results show that 52.5% of the 2,189 chemicals evaluated display hormetic concentration-responses in at least one of the 13 yeast strains. Many chemicals showed hormesis in multiple strains, and 24 agents showed hormesis in all 13 strains. The data are compared to previously reported quantitative features of hormesis based on published literature. PMID- 19543480 TI - Linear and non-linear dose-response functions reveal a hormetic relationship between stress and learning. AB - Over a century of behavioral research has shown that stress can enhance or impair learning and memory. In the present review, we have explored the complex effects of stress on cognition and propose that they are characterized by linear and non linear dose-response functions, which together reveal a hormetic relationship between stress and learning. We suggest that stress initially enhances hippocampal function, resulting from amygdala-induced excitation of hippocampal synaptic plasticity, as well as the excitatory effects of several neuromodulators, including corticosteroids, norepinephrine, corticotropin releasing hormone, acetylcholine and dopamine. We propose that this rapid activation of the amygdala-hippocampus brain memory system results in a linear dose-response relation between emotional strength and memory formation. More prolonged stress, however, leads to an inhibition of hippocampal function, which can be attributed to compensatory cellular responses that protect hippocampal neurons from excitotoxicity. This inhibition of hippocampal functioning in response to prolonged stress is potentially relevant to the well-described curvilinear dose-response relationship between arousal and memory. Our emphasis on the temporal features of stress-brain interactions addresses how stress can activate, as well as impair, hippocampal functioning to produce a hormetic relationship between stress and learning. PMID- 19543483 TI - Aspects of the relationship between drug dose and drug effect. AB - It is generally assumed that there exists a well-defined relationship between drug dose and drug effect and that this can be expressed by a dose-response curve. This paper argues that there is no such clear relation and that the dose response curve provides only limited information about the drug effect. It is demonstrated that tolerance development during the measurement of the dose response curve may cause major distortion of the curve and it is argued that the curve may only be used to indicate the response to the first administration of a drug, before tolerance has developed. The precise effect of a drug on an individual depends on the dynamic relation between several variables, particularly the level of tolerance, the dose anticipated by the organism and the actual drug dose. Simulations with a previously published mathematical model of drug tolerance demonstrate that the effect of a dose smaller than the dose the organism has developed tolerance to is difficult to predict and may be opposite to the action of the usual dose. PMID- 19543485 TI - Clinical outcome of paraquat poisoning. PMID- 19543486 TI - Marked recovery from paraquat-induced lung injury during long-term follow-up. AB - BACKGROUND/AIMS: Paraquat-induced lung injury has been considered a progressive and irreversible disease. The purpose of this study was to report the long-term evolution of lung lesions in eight survivors with significant paraquat-induced lung injuries who could be followed-up for longer than 6 months. METHODS: We retrospectively examined high-resolution computed tomography and pulmonary function test of eight survivors with significant paraquat-induced lung injurys. RESULTS: High-resolution computed tomography revealed a predominant pattern of irregularly shaped consolidation with traction bronchiectasis at 1-2 months after paraquat poisoning, a mixed pattern of irregularly shaped consolidation and ground-glass opacity at 3-12 months, and a mixed pattern of consolidation, ground glass opacity, and honeycombing at 1-2 years. At 3-12 months after paraquat ingestion, the areas of consolidation had markedly decreased and the decreased lung volume had returned to normal. At 1-2 years after paraquat poisoning, the cystic changes had disappeared. At 2-3 years after paraquat poisoning, the decrease in forced vital capacity had greatly improved to the normal range. CONCLUSIONS: Recovery of nearly normal pulmonary structure and function may occur over several years following paraquat poisoning. Pulmonary function (both forced vital capacity and forced expiratory volume in 1 sec) evolved toward normal in the long-term survivors of paraquat poisoning with initial prominent lung injuries. PMID- 19543484 TI - Established and newly proposed mechanisms of chronic cyclosporine nephropathy. AB - Cyclosporine (CsA) has improved patient and graft survival rates following solid organ transplantation and has shown significant clinical benefits in the management of autoimmune diseases. However, the clinical use of CsA is often limited by acute or chronic nephropathy, which remains a major problem. Acute nephropathy depends on the dosage of CsA and appears to be caused by a reduction in renal blood flow related to afferent arteriolar vasoconstriction. However, the mechanisms underlying chronic CsA nephropathy are not completely understood. Activation of the intrarenal renin-angiotensin system (RAS), increased release of endothelin-1, dysregulation of nitric oxide (NO) and NO synthase, up-regulation of transforming growth factor-beta1 (TGF-beta1), inappropriate apoptosis, stimulation of inflammatory mediators, enhanced innate immunity, endoplasmic reticulum stress, and autophagy have all been implicated in the pathogenesis of chronic CsA nephropathy. Reducing the CsA dosage or using other renoprotective drugs (angiotensin II receptor antagonist, mycophenolate mofetil, and statins, etc.) may ameliorate chronic CsA-induced renal injury. This review discusses old and new concepts in CsA nephropathy and preventive strategies for this clinical dilemma. PMID- 19543488 TI - The association between the serum sodium level and the severity of complications in liver cirrhosis. AB - BACKGROUND/AIMS: Dilutional hyponatremia associated with liver cirrhosis is caused by impaired free water clearance. Several studies have shown that serum sodium levels correlate with survival in cirrhotic patients. Little is known, however, regarding the relationship between the degree of dilutional hyponatremia and development of cirrhotic complications. The aim of this study was to evaluate the association between the serum sodium level and the severity of complications in liver cirrhosis. METHODS: Data of inpatients with cirrhotic complications were collected retrospectively. The serum sodium levels and severity of complications of 188 inpatients were analyzed. RESULTS: The prevalence of dilutional hyponatremia, classified as serum sodium concentrations of 0.174 ng/mL (best cut-off value) was considered positive for a diagnosis of bacterial PPE (sensitivity, 80%; specificity, 76%; AUC, 0.84). Pleural effusion PCT in the bacterial PPE is significantly different from those of the non-bacterial PPE and control groups, so the diagnostic use of PCT still warrants further investigation. PMID- 19543501 TI - Effects of mixed herbal extracts from parched Puerariae radix, gingered Magnoliae cortex, Glycyrrhizae radix and Euphorbiae radix (KIOM-79) on cardiac ion channels and action potentials. AB - KIOM-79, a mixture of ethanol extracts from four herbs (parched Puerariae radix, gingered Magnoliae cortex, Glycyrrhizae radix and Euphorbiae radix), has been developed for the potential therapeutic application to diabetic symptoms. Because screening of unexpected cardiac arrhythmia is compulsory for the new drug development, we investigated the effects of KIOM-79 on the action potential (AP) and various ion channel currents in cardiac myocytes. KIOM-79 decreased the upstroke velocity (V(max)) and plateau potential while slightly increased the duration of action potential (APD). Consistent with the decreased V(max) and plateau potential, the peak amplitude of Na+ current (I(Na)) and Ca2+ current (I(Ca,L)) were decreased by KIOM-79. KIOM-79 showed dual effects on hERG K+ current; increase of depolarization phase current (I(depol)) and decreased tail current at repolarization phase (I(tail)). The increase of APD was suspected due to the decreased I(tail). In computer simulation, the change of cardiac action potential could be well simulated based on the effects of KIOM-79 on various membrane currents. As a whole, the influence of KIOM-79 on cardiac ion channels are minor at concentrations effective for the diabetic models (0.1-10 microg/mL). The results suggest safety in terms of the risk of cardiac arrhythmia. Also, our study demonstrates the usefulness of the cardiac computer simulation in screening drug-induced long-QT syndrome. PMID- 19543502 TI - Ocular sarcoidosis in a Korean population. AB - The aim of current study was to evaluate the incidence and characteristics of ocular sarcoidosis in a Korean population. We conducted a retrospective study of 104 consecutive patients with biopsy-proven sarcoidosis seen at Asan Medical Center in Seoul, Korea, from 1993 to 2007. Medical records, photographs, and fluorescein angiograms were reviewed. Of 104 patients, 22 (21%) had intraocular involvement with female predominance (86%, M:F=3:19). Of the 39 eyes with ocular involvement, 16 (41%) eyes had isolated anterior uveitis, 12 (31%) eyes had intermediate uveitis, 6 eyes (15%) had panuveitis with retinal vasculitis, and 5 (13%) eyes had panuveitis with punched multifocal choroiditis. Mean duration of ophthalmologic follow-up was 62 months. All ocular inflammation was well managed with topical steroid and/or systemic steroid with relatively good final visual outcomes. Ocular complications such as cataract (12 eyes, 30%), glaucoma (6 eyes, 15%), vitreous opacity (1 eye, 3%), cystoid macular edema (3 eyes, 7%), neovascularization (2 eye, 5%), and epiretinal membrane (4 eye, 10%) were related to ocular sarcoidosis. In Korea, where sarcoidosis is very rare, our study indicates relatively low ocular and predominantly non posterior segment involvement with relatively good visual prognosis. PMID- 19543504 TI - Comparison of clinical and radiographic characteristics between nodular bronchiectatic form of nontuberculous mycobacterial lung disease and diffuse panbronchiolitis. AB - The nodular bronchiectatic form of nontuberculous mycobacterial (NTM) lung disease and diffuse panbronchiolits (DPB) show similar clinical and radiographic findings. The present study was performed to clarify the clinicoradiographic similarities as well as the differences between NTM lung disease and DPB. The initial clinicoradiographic features of 78 patients with the nodular bronchiectatic form of NTM lung disease (41 patients with Mycobacterium avium complex infection and 37 patients with Mycobacterium abscessus infection) were compared with those of 35 patients with DPB. Old age, female sex, a history of tuberculosis treatment, and hemoptysis were related to NTM lung disease while exertional dyspnea, coarse crackles, history of sinusitis, obstructive abnormalities in pulmonary function tests, and hypoxemia were related to DPB. The number of lobes involved with bronchiolitis and bronchiectasis on chest computed tomography were more numerous in DPB patients. There is considerable overlap in the clinical and radiographic appearances of the nodular bronchiectatic form of NTM lung disease and DPB, although some clinicoradiographic features differ between two diseases. The correct diagnosis, including aggressive microbiologic evaluation, should be made for the appropriate management of patients presenting with bilateral bronchiectasis and bronchiolitis. PMID- 19543505 TI - ACVR1 gene mutation in sporadic Korean patients with fibrodysplasia ossificans progressiva. AB - Fibrodysplasia ossificans progressiva (FOP; OMIM 135100) is a rare but extremely disabling genetic disorder of the skeletal system, and is characterized by the progressive development of ectopic ossification of skeletal muscles and subsequent joint ankylosis. The c.617G>A; p.R206H point mutation in the activin A type I receptor (ACVR1) gene has been reported to be a causative mutation of FOP. In the present study, mutation analysis of the ACVR1 gene was performed in 12 patients diagnosed or suspected to have FOP. All patients tested had a de novo heterozygous point mutation of c.617G>A; p.R206H in ACVR1. Mutation analysis confirmed a diagnosis of FOP in patients with ambiguous features, and thus, could be used for diagnostic purposes. Early confirmation through mutation analysis would allow medical professionals to advise on the avoidance of provoking events to delay catastrophic flare-ups of ectopic ossifications. PMID- 19543503 TI - Acute Physiology and Chronic Health Evaluation II and Simplified Acute Physiology Score II in predicting hospital mortality of neurosurgical intensive care unit patients. AB - We study the predictive power of Acute Physiology and Chronic Health Evaluation II (APACHE II) and Simplified Acute Physiology Score II (SAPS II) in neurosurgical intensive care unit (ICU) patients. Retrospective investigation was conducted on 672 consecutive ICU patients during the last 2 yr. Data were collected during the first 24 hours of admission and analyzed to calculate predicted mortality. Mortality predicted by two systems was compared and, multivariate analyses were then performed for subarachnoid hemorrhage (SAH) and traumatic brain injury (TBI) patients. Observed mortality was 24.8% whereas predicted mortalities were 37.7% and 38.4%, according to APACHE II and SAPS II. Calibration curve was close to the line of perfect prediction. SAPS II was not statistically significant according to a Lemeshow-Hosmer test, but slightly favored by area under the curve (AUC). In SAH patients, SAPS II was an independent predictor for mortality. In TBI patients, both systems had independent prognostic implications. Scoring systems are useful in predicting mortality and measuring performance in neurosurgical ICU setting. TBI patients are more affected by systemic insults than SAH patients, and this discrepancy of predicting mortality in each neurosurgical disease prompts us to develop a more specific scoring system targeted to cerebral dysfunction. PMID- 19543506 TI - Association of interleukin-10 A-592C polymorphism in Taiwanese children with Kawasaki disease. AB - Elevated serum levels of interleukin-10 (IL-10) have been reported in patients with Kawasaki disease (KD). IL-10 reduces the inflammatory actions of macrophages and T cells and it may play a significant role in the regulation of inflammatory vascular damage associated with systemic vasculitis. The aim of this study was to examine whether -592 IL-10 promoter polymorphism is a susceptibility or severity marker of KD in Chinese patients in Taiwan. The study included 105 KD patients and 100 normal controls. Genotype and allelic frequencies for the IL-10 gene polymorphism in both groups were compared. There were no significant between group differences in the genotype distribution of IL-10 A-592C gene polymorphism (P=0.08). However, the frequency of the -592*A allele was significantly increased in the patients with KD compared with controls (71.9% vs. 61.0%, P=0.019). The odds ratio for developing KD in individuals with IL-10-592*A allele was 1.64 (95% confidence interval, 1.06-2.52) compared to individuals with the IL-10-592*C allele. No significant difference was observed in the genotype and allelic frequencies for the IL-10 A-592C polymorphism between patients with and without coronary artery lesions. The IL-10-592*A allele may be involved in the development of KD in Taiwanese children. PMID- 19543507 TI - Non-association between polymorphisms of the frizzled receptor genes and bone mineral density in postmenopausal Korean women. AB - We investigated the association between single nucleotide polymorphisms (SNPs) in the frizzled (FZD) genes in the Wnt signal pathway and circulating osteoprotegerin (OPG), soluble receptor activator of NF-kappaB ligand (sRANKL) levels, bone turnover markers, and bone mineral density (BMD) in postmenopausal women. The SNPs in the FZD1, FZD5, FZD6, FZD7, and FZD9 genes were analyzed by direct sequencing in 371 postmenopausal Korean women. Levels of serum OPG, sRANKL, osteocalcin, C-telopeptide of type I collagen, calcium, parathyroid hormone and calcitonin, and BMD at the lumbar spine and femoral neck were measured. The SNPs in the FZD1, FZD5, FZD7, and FZD9 genes, and in exon 2 of the FZD6 gene were not observed. No significant differences in the adjusted BMD of lumbar spine and femoral neck and serum levels of OPG, sRANKL, and bone markers were noted among the single or haplotype genotypes of the L345M and E664A SNPs in the FZD6 gene and the distributions of these single or haplotype genotypes were not different according to the bone mass status. In conclusion, the polymorphisms of the FZD genes are not associated with BMD of the lumbar spine and femoral neck, bone turnover markers, or circulating OPG-sRANKL in Korean women. PMID- 19543508 TI - Epidermal growth factor receptor mutations and the clinical outcome in male smokers with squamous cell carcinoma of lung. AB - Epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) have been reported to be related to certain clinical characteristics (i.e., female, non-smokers with adenocarcinoma) and gefitinib responsiveness. This exploratory analysis was performed to determine the incidence of EGFR mutations in male smokers with squamous cell carcinoma, who were treated with EGFR tyrosine kinase inhibitor, gefitinib. Sixty-nine Korean NSCLC patients were treated with gefitinib in a prospective study. For a subset of 20 male patients with squamous cell carcinoma and a history of smoking, pretreatment tumor tissue samples were obtained and analyzed for EGFR mutations (exons 18 to 21). EGFR mutations were found in 3 (15%) patients, including in-frame deletions within exon 19 (n=2) and L858R missence mutation in exon 21 (n=1). These 3 patients with EGFR mutations responded to gefitinib, whereas only one of remaining 17 patients with wild-type EGFR achieved clinical response. Trend toward longer progression free (5.8 vs. 2.4 months; P=0.07) was noted in patients with EGFR mutations compared to those with wild-type EGFR. Although male smokers with squamous cell carcinoma have not been considered ideal candidates for gefitinib treatment, significant incidence of EGFR mutations was observed. The molecular markers should be considered to predict clinical benefits from gefitinib. PMID- 19543509 TI - Cytogenetic characterizations of central nervous system tumors: the first comprehensive report from a single institution in Korea. AB - The World Health Organization (WHO) classification of central nervous system (CNS) tumors incorporates morphology, cytogenetics, molecular genetics, and immunologic markers. Despite the relatively large number of CNS tumors with clonal chromosome abnormalities, only few studies have investigated cytogenetic abnormalities for CNS tumors in Korea. Thus, we investigated 119 CNS tumors by conventional G-banded karyotypes to characterize patterns of chromosomal abnormalities involving various CNS tumors, and 92.4% of them were cultured and karyotyped successfully. Totally, 51.8% of karyotypable CNS tumors showed abnormal cytogenetic results, including neuroepithelial tumors (75.0%), meningeal tumors (71.1%), pituitary adenomas (4.2%), schwannomas (44.4%), and metastatic tumors (100.0%). Glioblastomas had hyperdiploid, complex karyotypes, mainly involving chromosomes Y, 1, 2, 6, 7, 10, 12, 13, and 14. Monosomy 22 was observed in 56.4% of meningiomas. There was a significant increase in the frequencies of karyotypic complexity according to the increase of WHO grade between grades I and II (P=0.0422) or IV (P=0.0101). Abnormal karyotypes were more complex at high grade tumors, suggesting that the karyotype reflects the biologic nature of the tumor. More detailed cytogenetic and molecular characterizations of CNS tumors contribute to better diagnostic criteria and deeper insights of tumorigenesis, eventually resulting in development of novel therapeutic strategies. PMID- 19543510 TI - Neuroblastoma originating from extra-abdominal sites: association with favorable clinical and biological features. AB - Neuroblastomas originating from different sites might have different clinical and biological characteristics. In the present study, the clinical (age, sex and stage) and biological (N-myc amplification, Shimada pathology and levels of lactate dehydrogenase, ferritin and neuron-specific enolase) characteristics of patients with newly diagnosed neuroblastoma were compared according to the site of tumor origin (extra-abdominal versus abdominal). The event-free survival rate (EFS) was also compared between the two groups. Among 143 neuroblastomas, 115 tumors originated from the abdomen, 26 from extra-abdominal sites and 2 from unknown primary sites. Frequencies of stage 4 tumor and N-myc amplified tumor were lower in the extra-abdominal group than in the abdominal group (34.6% vs. 60.0%, P=0.019 and 4.2% vs. 45.0%, P<0.001, respectively). Levels of lactate dehydrogenase, ferritin and neuron-specific enolase were significantly lower in the extra-abdominal group than in the abdominal group. The probability of 5-yr EFS (+/-95% confidence interval) was higher in the extra-abdominal group than in the abdominal group (94.4+/-10.6% vs. 69.4+/-9.4%, P=0.026). Taken together, neuroblastomas originating from extra-abdominal sites might be associated with more favorable clinical and biological characteristics and a better outcome than neuroblastomas originating from abdomen. PMID- 19543511 TI - Forkhead box O-class 1 and forkhead box G1 as prognostic markers for bladder cancer. AB - Forkhead box O-class 1 (FOXO1) is a key regulator of glucose homeostasis, cell cycle progression, and apoptosis. Its functions are modulated by forkhead box G1 (FOXG1), which acts as a transcriptional repressor with oncogenic potential. Real time PCR and immunohistochemical staining were performed in 174 primary bladder cancer specimens and 21 normal bladder mucosae to evaluate these genes. FOXO1 and FOXG1 mRNA expression in cancer tissues were higher than in normal mucosae (each P<0.001). FOXO1 mRNA levels were significantly higher in samples of non progressed patients (P<0.001), but FOXG1 were enhanced in those of progressed patients (P=0.019). On univariate analysis, FOXO1 mRNA expression was significantly associated with grade, stage, recurrence, progression and survival (each P<0.05). On multivariate analysis, increased FOXO1 mRNA expression was associated with both reduced disease progression (odds ratio [OR], 0.367; 95% confidence interval [CI], 0.163-0.826, P=0.015) and enhanced disease-free survival (OR, 3.262; 95% CI, 1.361-7.820, P=0.008). At a median follow-up of 33 months (range 2 to 156), the patients with a high FOXO1 mRNA expression had a significantly prolonged survival (P=0.001). Immunohistochemical findings of FOXO1 were generally concordant with mRNA expression levels. In conclusion, FOXO1 may be a promising marker for predicting progression in human bladder cancers. PMID- 19543512 TI - EC-18, a synthetic monoacetyldiacylglyceride, inhibits hematogenous metastasis of KIGB-5 biliary cancer cell in hamster model. AB - EC-18 (monoacetyldiacylglyceride) stimulates T cell production of IL-2, IL-4, IL 12, IFN-gamma, and GM-CSF in vitro. To study the effects of these cytokines stimulated by EC-18 on cancer cells, we applied hamster biliary cancer model, a difficult cancer to treat. Cancer (KIGB-5) cells were given intravenously to produce hematogenous metastatic lung lesions which were treated with EC-18 at 10, 25, and 50 mg/kg/day respectively. The fourth group was untreated control. At 4th, 8th, and 12th week the lungs were examined. EC-18 treated groups showed only a few microscopic lung lesions and no evidence of metastatic lesion with highest dose whereas widespread gross lung lesions were observed in untreated control. To investigate whether the anti-tumor effect of EC-18 is associated with suppression of tumor cell Toll-like receptor 4 (TLR-4) expression in addition to stimulation of the immune cells, KIGB-5 cells were exposed to LPS with or without EC-18. TLR 4 mRNA and protein expression, measured by reverse transcriptase PCR (RT-PCR), real-time quantitative PCR and western blot analysis, showed suppression of TLR-4 expression in KIGB-5 cells treated with EC-18 compared with control. In conclusion, EC-18 has a significant anti-tumor effect in this experimental model of biliary cancer suggesting potential for clinical application to this difficult cancer. PMID- 19543513 TI - Clinicopathologic features of polypoid lesions of the gallbladder and risk factors of gallbladder cancer. AB - It is difficult to differentiate benign and malignancy in polypoid lesions of the gallbladder (PLG) by solely depending on imaging studies. Therefore clinicopathologic features of benign and malignant polyps are compared in an attempt to identify the risk factors of malignant polypoid lesions. The medical records of 291 patients who were confirmed to have PLG through cholecystectomy were reviewed and analyzed for age, sex, symptom, associated gallstone, morphology of PLG, size of PLG, number of PLG, and preoperative tumor markers. Benign PLG was found in 256 patients (88.0%) and malignant PLG in 35 patients (12.0%). Compared with benign group, the malignant group were older (61.1 yr vs. 47.1 yr, P<0.001), more often accompanied with symptoms (62.9% vs. 28.9%, P<0.001). Malignant PLG tended to be sessile (60.0% vs. 10.5%, P<0.001), larger (28.0 mm vs. 8.6 mm, P<0.001) and single lesion (65.7% vs. 44.1%, P<0.016). Age over 60 yr (P=0.021, odds ratio [OR], 8.16), sessile morphology (P<0.001, OR, 7.70), and size over 10 mm (P=0.009, OR, 8.87) were identified as risk factors for malignant PLG. Careful decision making on therapeutic plans should be made with consideration of malignancy for patients over 60 yr, with sessile morphology of PLG, and with PLG size of over 10 mm. PMID- 19543514 TI - Stereotactic body radiotherapy for isolated para-aortic lymph node recurrence after curative resection in gastric cancer. AB - The aim of this study was to investigate whether stereotactic body radiotherapy (SBRT) can salvage gastric cancer patients with para-aortic lymph node (PALN) recurrence. From January 2003 to December 2006, 7 patients were treated for isolated PALN recurrence from gastric cancer after curative resection. Follow up durations ranged from 19 to 33 months (median; 26 months), and SBRT doses from 45 Gy to 51 Gy (median 48 Gy) in 3 fractions. Disease progression-free and overall survivals and toxicities were recorded. Response to treatment was assessed by computed tomography. Final patient outcomes were as follows: 2 were alive without evidence of disease, 3 remained alive with disease, and 2 patients died of disease. Five of 7 patients showed complete response and 2 patients partial response between 3 and 11 months after SBRT. Three-year overall and disease progression-free survival rates post-SBRT were 43% and 29%, respectively. No severe complication was detected during follow-up. Selected patients with isolated PALN recurrence can be salvaged by SBRT without severe complications. PMID- 19543515 TI - Differential methylation pattern of ID4, SFRP1, and SHP1 between acute myeloid leukemia and chronic myeloid leukemia. AB - To gain insight into the differential mechanism of gene promoter hypermethylation in acute and chronic leukemia, we identified the methylation status on one part of 5'CpG rich region of 8 genes, DAB2IP, DLC-1, H-cadherin, ID4, Integrin alpha4, RUNX3, SFRP1, and SHP1 in bone marrows from acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) patients. Also, we compared the methylation status of genes in AML and CML using methylation-specific PCR (MSP). The frequencies of DNA methylation of ID4, SFRP1, and SHP1 were higher in AML patients compared to those in CML patients. In contrast, no statistical difference between AML and CML was detected for other genes such as DLC-1, DAB2IP, H-cadherin, Integrin alpha4, and RUNX3. Taken together, these results suggest that these methylation controlled genes may have different roles in AML and CML, and thus, may act as a biological marker of AML. PMID- 19543516 TI - The clinical outcome of FLAG chemotherapy without idarubicin in patients with relapsed or refractory acute myeloid leukemia. AB - A refractory and resistant disease to conventional induction chemotherapy and relapsed disease are considered as the most important adverse prognostic factors for acute myeloid leukemia (AML). Sixty-one patients (median age, 33.6 yr) with relapsed or refractory AML were treated with the FLAG regimen that consisted of fludarabine (30 mg/m(2), days 1-5), cytarabine (2.0 g/m(2), days 1-5) and granulocyte colony-stimulating factor. Of the treated patients 29 patients (47.5%) achieved complete remission (CR). Higher CR rates were observed for patients with a first or second relapse as compared to patients with a primary refractory response or relapse after stem cell transplantation (HSCT). There was a significant difference in the response rates according to the duration of leukemia-free survival (pre-LFS) before chemotherapy (P=0.05). The recovery time of both neutrophils (> or =500/microL) and platelets (> or =20,000/microL) required a median of 21 and 18 days, respectively. Treatment-related mortality (TRM) occurred in seven patients (11.4%), of which 71.4% of TRM was caused by an invasive aspergillosis infection. After achieving CR, 18 patients underwent consolidation chemotherapy and six patients underwent allogeneic HSCT. In conclusion, FLAG chemotherapy without idarubicin is a relatively effective and well-tolerated regimen for relapsed or refractory AML and the use of FLAG chemotherapy has allowed intensive post-remission therapy including HSCT. PMID- 19543517 TI - Pulmonary adenocarcinoma with heterotopic ossification. AB - Pulmonary adenocarcinoma is a common malignancy that often involves calcification; however, bone formation in primary lung adenocarcinoma is extremely rare. In ten cases of primary pulmonary adenocarcinoma with heterotopic ossification, we detected immunoreactivity against TGF-beta1, osteopontin, osteocalcin and Runx2 in the fibroblastic stroma and tumor cells within the area of ossification. Our results suggest that in primary pulmonary adenocarcinoma, heterotopic ossification occurs via intramembranous bone formation. To our knowledge, only 11 other cases of pulmonary adenocarcinoma with heterotopic ossification have been reported. Here, we present ten cases of pulmonary adenocarcinoma showing heterotopic ossification with a description of previously published results and the histogenesis of heterotopic bone formation. PMID- 19543518 TI - Clarithromycin susceptibility testing of Mycobacterium avium complex using 2,3 diphenyl-5-thienyl-(2)-tetrazolium chloride microplate assay with Middlebrook 7H9 broth. AB - A series of 119 Mycobacterium avium complex isolates were subjected to clarithromycin susceptibility testing using microplates containing 2,3-diphenyl-5 thienyl-(2)-tetrazolium chloride (STC). Among 119 isolates, 114 (95.8%) were susceptible to clarithromycin and 5 were resistant according to the new and the standard method. STC counts the low cost and reduces the number of procedures needed for susceptibility testing. PMID- 19543519 TI - A case of successful selective abortion using radio-frequency ablation in twin pregnancy suffering from severe twin to twin transfusion syndrome. AB - Twin to twin transfusion syndrome (TTTS) is one of the major complication of monochorionic twin pregnancy which is mainly understood by placental vascular anastomosis. Perinatal mortality and morbidity is high as 80-100% if untreated and even higher if the disease is developed at early stage. Variety of methods of isolating or intercepting placental vascular anastomosis are introduced, but they are only available in centers where all the required equipments are prepared. We report here a case of TTTS complicated with severe polyhydroamnios during the second trimester. The blood supply to donor twin was interrupted successfully at 19(+2) weeks of gestation by minimally invasive radio-frequency cord ablation, under ultrasound guidance. The normal recipient twin was delivered successfully at 35 weeks of gestation and had no eventful neonatal course. PMID- 19543520 TI - Congenital hemidiaphragmatic agenesis presenting as reversible mesenteroaxial gastric volvulus and diaphragmatic hernia: a case report. AB - A 70-yr-old woman complained of left sided chest pain and non-bilious vomiting for four days after taking a gastric bloating agent for an upper gastrointestinal study. The chest radiography revealed gastric air-fluid levels and bowel loops in the left thoracic cavity. An emergency thoracotomy was performed. The abdominal organs (stomach, spleen, splenic flexure of the colon) were in the left thorax and the entire left hemidiaphragm was absent. There were no diaphragmatic remnants visible for reconstruction of the left diaphragm. We provided warm saline irrigation and performed a left lower lobe adhesiotomy. Thirteen days after surgery, the chest radiography showed improvement in the herniation but mild haziness remained at the left lower lung field. Here we present the oldest case of congenital diaphragmatic agenesis presenting with transient gastric volvulus and diaphragmatic hernia. PMID- 19543521 TI - The first case of antibiotic-associated colitis by Clostridium difficile PCR ribotype 027 in Korea. AB - Clostridium difficile (C. difficile) is a common causative agent of pseudomembranous colitis (PMC). C. difficile-associated diarrhea (CDAD) ranges from mild diarrhea to life threatening PMC. Recently, a highly virulent strain of C. difficile polymerase chain reaction ribotype 027 was found in North America, Europe, and Japan. A 52-yr-old woman with anti-tuberculosis medication and neurogenic bladder due to traffic accident experienced five episodes of C. difficile PMC after taking antibiotics for pneumonia along with septic shock and acute renal failure. She was readmitted to the intensive care unit and treated with oral vancomycin with refractory of oral metronidazole, inotropics and probiotics for over 60 days. C. difficile isolated both at the first and the last admission was identified as C. difficile ribotype 027 by ribotyping, toxinotyping, and tcdC gene sequencing, which turned out the same pathogen as the epidemic hypervirulent B1/NAP1 strain. This is the first case of C. difficile PCR ribotype 027 in Korea. After discharge, she was maintained on probiotics and rifaximin for 3 weeks. She had no relapse for 6 months. PMID- 19543522 TI - A case of primary adrenal diffuse large B-cell lymphoma achieving complete remission with rituximab-CHOP chemotherapy. AB - Primary adrenal lymphoma is a very rare extranodal lymphoma; its clinical features consist of a high incidence of bilateral adrenal involvement and diffuse large B-cell lymphoma. We report a patient with primary bilateral adrenal diffuse large B-cell lymphoma who achieved complete remission with R-CHOP (rituximab cyclophosphamide, doxorubicin, vincristine, and prednisone) chemotherapy. A 52-yr old man presented with fever and progressive fatigue for 3 months. Computed tomography (CT) scans of the abdomen and pelvis demonstrated large bilateral adrenal masses, and a needle biopsy of the left adrenal mass revealed diffuse large B-cell lymphoma. After 6 cycles of R-CHOP chemotherapy, CT scans showed no residual disease. To our knowledge, this is the second report to date of a patient with primary bilateral adrenal diffuse large B-cell lymphoma who achieved complete remission using R-CHOP chemotherapy. PMID- 19543523 TI - Endometrial stromal sarcoma presenting as prevesical mass mimicking urachal tumor. AB - Endometrial stromal sarcoma (ESS) is a mesenchymal neoplasm that usually occurs as a primary tumor of the uterine corpus, but rarely arises in other sites, such as the ovary, pelvic cavity, mesentery, omentum and intestine. Herein, we present a rare case of low-grade ESS presented as prevesical mass. A 60-yr-old woman who had undergone total hysterectomy for endometriosis eleven years ago was presented with incidentally detected prevesical pelvic mass. Since malignant transformation of urachal remnants was possible, the mass was suspected to be a urachal tumor. Extraction of the mass was performed, and the histopathologic diagnosis was low grade ESS. In summary, prevesical tumor is rare but in patients with endometriosis, we suggest endometriosis and its possible malignant changes should be taken into account in the differential diagnosis of prevesical mass. PMID- 19543524 TI - 20-Hydroxycholecalciferol, product of vitamin D3 hydroxylation by P450scc, decreases NF-kappaB activity by increasing IkappaB alpha levels in human keratinocytes. AB - The side chain of vitamin D3 is hydroxylated in a sequential manner by cytochrome P450scc (CYP11A1) to form 20-hydroxycholecalciferol, which can induce growth arrest and differentiation of both primary and immortalized epidermal keratinocytes. Since nuclear factor-kappaB (NF-kappaB) plays a pivotal role in the regulation of cell proliferation, differentiation and apoptosis, we examined the capability of 20-hydroxycholecalciferol to modulate the activity of NF kappaB, using 1,25-dihydroxycholecalciferol (calcitriol) as a positive control. 20-hydroxycholecalciferol inhibits the activation of NFkappaB DNA binding activity as well as NF-kappaB-driven reporter gene activity in keratinocytes. Also, 20-hydroxycholecalciferol induced significant increases in the mRNA and protein levels of the NF-kappaB inhibitor protein, IkappaB alpha, in a time dependent manner, while no changes in total NF-kappaB-p65 mRNA or protein levels were observed. Another measure of NF-kappaB activity, p65 translocation from the cytoplasm into the nucleus was also inhibited in extracts of 20 hydroxycholecalciferol treated keratinocytes. Increased IkappaB alpha was concomitantly observed in cytosolic extracts of 20-hydroxycholecalciferol treated keratinocytes, as determined by immunoblotting and immunofluorescent staining. In keratinocytes lacking vitamin D receptor (VDR), 20-hydroxycholecalciferol did not affect IkappaB alpha mRNA levels, indicating that it requires VDR for its action on NF-kappaB activity. Comparison of the effects of calcitrol, hormonally active form of vitamin D3, with 20-hydrocholecalciferol show that both agents have a similar potency in inhibiting NF-kappaB. Since NF-kappaB is a major transcription factor for the induction of inflammatory mediators, our findings indicate that 20 hydroxycholecalciferol may be an effective therapeutic agent for inflammatory and hyperproliferative skin diseases. PMID- 19543525 TI - Shared visual attention and memory systems in the Drosophila brain. AB - BACKGROUND: Selective attention and memory seem to be related in human experience. This appears to be the case as well in simple model organisms such as the fly Drosophila melanogaster. Mutations affecting olfactory and visual memory formation in Drosophila, such as in dunce and rutabaga, also affect short-term visual processes relevant to selective attention. In particular, increased optomotor responsiveness appears to be predictive of visual attention defects in these mutants. METHODOLOGY/PRINCIPAL FINDINGS: To further explore the possible overlap between memory and visual attention systems in the fly brain, we screened a panel of 36 olfactory long term memory (LTM) mutants for visual attention-like defects using an optomotor maze paradigm. Three of these mutants yielded high dunce-like optomotor responsiveness. We characterized these three strains by examining their visual distraction in the maze, their visual learning capabilities, and their brain activity responses to visual novelty. We found that one of these mutants, D0067, was almost completely identical to dunce(1) for all measures, while another, D0264, was more like wild type. Exploiting the fact that the LTM mutants are also Gal4 enhancer traps, we explored the sufficiency for the cells subserved by these elements to rescue dunce attention defects and found overlap at the level of the mushroom bodies. Finally, we demonstrate that control of synaptic function in these Gal4 expressing cells specifically modulates a 20 30 Hz local field potential associated with attention-like effects in the fly brain. CONCLUSIONS/SIGNIFICANCE: Our study uncovers genetic and neuroanatomical systems in the fly brain affecting both visual attention and odor memory phenotypes. A common component to these systems appears to be the mushroom bodies, brain structures which have been traditionally associated with odor learning but which we propose might be also involved in generating oscillatory brain activity required for attention-like processes in the fly brain. PMID- 19543526 TI - Immune regulatory neural stem/precursor cells protect from central nervous system autoimmunity by restraining dendritic cell function. AB - BACKGROUND: The systemic injection of neural stem/precursor cells (NPCs) provides remarkable amelioration of the clinico-pathological features of experimental autoimmune encephalomyelitis (EAE). This is dependent on the capacity of transplanted NPCs to engage concurrent mechanisms of action within specific microenvironments in vivo. Among a wide range of therapeutic actions alternative to cell replacement, neuroprotective and immune modulatory capacities of transplanted NPCs have been described. However, lacking is a detailed understanding of the mechanisms by which NPCs exert their therapeutic plasticity. This study was designed to identify the first candidate that exemplifies and sustains the immune modulatory capacity of transplanted NPCs. METHODOLOGY/PRINCIPAL FINDINGS: To achieve the exclusive targeting of the peripheral immune system, SJL mice with PLP-induced EAE were injected subcutaneously with NPCs and the treatment commenced prior to disease onset. NPC injected EAE mice showed significant clinical improvement, as compared to controls. Exogenous NPCs lacking the expression of major neural antigens were reliably (and for long-term) found at the level of draining lymph nodes, while establishing sophisticated anatomical interactions with lymph node cells. Importantly, injected NPCs were never found in organs other than lymph nodes, including the brain and the spinal cord. Draining lymph nodes from transplanted mice showed focal up-regulation of major developmental stem cell regulators, such as BMP-4, Noggin and Sonic hedgehog. In lymph nodes, injected NPCs hampered the activation of myeloid dendritic cells (DCs) and steadily restrained the expansion of antigen-specific encephalitogenic T cells. Both ex vivo and in vitro experiments identified a novel highly NPC-specific-BMP-4-dependent-mechanism hindering the DC maturation. CONCLUSION/SIGNIFICANCE: The study described herein, identifies the first member of the TGF beta/BMP family of stem cell regulators as a novel tolerogenic factor released by NPCs. Full exploitation of this pathway as an efficient tool for vaccination therapy in autoimmune inflammatory conditions is underway. PMID- 19543528 TI - Association between common germline genetic variation in 94 candidate genes or regions and risks of invasive epithelial ovarian cancer. AB - BACKGROUND: Recent studies have identified several single nucleotide polymorphisms (SNPs) in the population that are associated with variations in the risks of many different diseases including cancers such as breast, prostate and colorectal. For ovarian cancer, the known highly penetrant susceptibility genes (BRCA1 and BRCA2) are probably responsible for only 40% of the excess familial ovarian cancer risks, suggesting that other susceptibility genes of lower penetrance exist. METHODS: We have taken a candidate approach to identifying moderate risk susceptibility alleles for ovarian cancer. To date, we have genotyped 340 SNPs from 94 candidate genes or regions, in up to 1,491 invasive epithelial ovarian cancer cases and 3,145 unaffected controls from three different population based studies from the UK, Denmark and USA. RESULTS: After adjusting for population stratification by genomic control, 18 SNPs (5.3%) were significant at the 5% level, and 5 SNPs (1.5%) were significant at the 1% level. The most significant association was for the SNP rs2107425, located on chromosome 11p15.5, which has previously been identified as a susceptibility allele for breast cancer from a genome wide association study (P-trend = 0.0012). When SNPs/genes were stratified into 7 different pathways or groups of validation SNPs, the breast cancer associated SNPs were the only group of SNPs that were significantly associated with ovarian cancer risk (P-heterogeneity = 0.0003; P trend = 0.0028; adjusted (for population stratification) P-trend = 0.006). We did not find statistically significant associations when the combined data for all SNPs were analysed using an admixture maximum likelihood (AML) experiment-wise test for association (P-heterogeneity = 0.051; P-trend = 0.068). CONCLUSION: These data suggest that a proportion of the SNPs we evaluated were associated with ovarian cancer risk, but that the effect sizes were too small to detect associations with individual SNPs. PMID- 19543527 TI - Non mycobacterial virulence genes in the genome of the emerging pathogen Mycobacterium abscessus. AB - Mycobacterium abscessus is an emerging rapidly growing mycobacterium (RGM) causing a pseudotuberculous lung disease to which patients with cystic fibrosis (CF) are particularly susceptible. We report here its complete genome sequence. The genome of M. abscessus (CIP 104536T) consists of a 5,067,172-bp circular chromosome including 4920 predicted coding sequences (CDS), an 81-kb full-length prophage and 5 IS elements, and a 23-kb mercury resistance plasmid almost identical to pMM23 from Mycobacterium marinum. The chromosome encodes many virulence proteins and virulence protein families absent or present in only small numbers in the model RGM species Mycobacterium smegmatis. Many of these proteins are encoded by genes belonging to a "mycobacterial" gene pool (e.g. PE and PPE proteins, MCE and YrbE proteins, lipoprotein LpqH precursors). However, many others (e.g. phospholipase C, MgtC, MsrA, ABC Fe(3+) transporter) appear to have been horizontally acquired from distantly related environmental bacteria with a high G+C content, mostly actinobacteria (e.g. Rhodococcus sp., Streptomyces sp.) and pseudomonads. We also identified several metabolic regions acquired from actinobacteria and pseudomonads (relating to phenazine biosynthesis, homogentisate catabolism, phenylacetic acid degradation, DNA degradation) not present in the M. smegmatis genome. Many of the "non mycobacterial" factors detected in M. abscessus are also present in two of the pathogens most frequently isolated from CF patients, Pseudomonas aeruginosa and Burkholderia cepacia. This study elucidates the genetic basis of the unique pathogenicity of M. abscessus among RGM, and raises the question of similar mechanisms of pathogenicity shared by unrelated organisms in CF patients. PMID- 19543529 TI - Rapid insulin-dependent endocytosis of the insulin receptor by caveolae in primary adipocytes. AB - BACKGROUND: The insulin receptor is localized in caveolae and is dependent on caveolae or cholesterol for signaling in adipocytes. When stimulated with insulin, the receptor is internalized. METHODOLOGY/PRINCIPAL FINDINGS: We examined primary rat adipocytes by subcellular fractionation to examine if the insulin receptor was internalized in a caveolae-mediated process. Insulin induced a rapid, t(1/2)<3 min, endocytosis of the insulin receptor in parallel with receptor tyrosine autophosphorylation. Concomitantly, caveolin-1 was phosphorylated at tyrosine(14) and endocytosed. Vanadate increased the phosphorylation of caveolin-1 without affecting insulin receptor phosphorylation or endocytosis. Immunocapture of endosomal vesicles with antibodies against the insulin receptor co-captured caveolin-1 and immunocapture with antibodies against tyrosine(14)-phosphorylated caveolin-1 co-captured the insulin receptor, demonstrating that the insulin receptor was endocytosed together with tyrosine(14)-phosphorylated caveolin-1. By immunogold electron microscopy the insulin receptor and caveolin-1 were colocalized in endosome vesicles that resembled caveosomes. Clathrin was not endocytosed with the insulin receptor and the inhibitor of clathrin-coated pit-mediated endocytosis, chlorpromazine, did not inhibit internalization of the insulin receptor, while transferrin receptor internalization was inhibited. CONCLUSION: It is concluded that in response to insulin stimulation the autophosphorylated insulin receptor in primary adipocytes is rapidly endocytosed in a caveolae-mediated process, involving tyrosine phosphorylation of caveolin-1. PMID- 19543530 TI - Plk1-dependent recruitment of gamma-tubulin complexes to mitotic centrosomes involves multiple PCM components. AB - The nucleation of microtubules requires protein complexes containing gamma tubulin, which are present in the cytoplasm and associate with the centrosome and with the mitotic spindle. We have previously shown that these interactions require the gamma-tubulin targeting factor GCP-WD/NEDD1, which has an essential role in spindle formation. The recruitment of additional gamma-tubulin to the centrosomes occurs during centrosome maturation at the G2/M transition and is regulated by the mitotic kinase Plk1. However, the molecular details of this important pathway are unknown and a Plk1 substrate that controls gamma-tubulin recruitment has not been identified. Here we show that Plk1 associates with GCP WD in mitosis and Plk1 activity contributes to phosphorylation of GCP-WD. Plk1 depletion or inhibition prevents accumulation of GCP-WD at mitotic centrosomes, but GCP-WD mutants that are defective in Plk1-binding and -phosphorylation still accumulate at mitotic centrosomes and recruit gamma-tubulin. Moreover, Plk1 also controls the recruitment of other PCM proteins implicated in centrosomal gamma tubulin attachment (Cep192/hSPD2, pericentrin, Cep215/Cdk5Rap2). Our results support a model in which Plk1-dependent recruitment of gamma-tubulin to mitotic centrosomes is regulated upstream of GCP-WD, involves multiple PCM proteins and therefore potentially multiple Plk1 substrates. PMID- 19543533 TI - Field evaluation of Calypte's AWARE blood serum plasma (BSP) and oral mucosal transudate (OMT) rapid tests for detecting antibodies to HIV-1 and 2 in plasma and oral fluid. AB - As programs to prevent and care for HIV-infected persons are scaled-up in Africa, there is the need for continuous evaluation of the performance of test kits that could best support these programs. The present study evaluated the sensitivity, specificity, ease of use, and cost of AWARE Blood Serum Plasma (BSP) and Oral Mucosal Transudate (OMT) Rapid HIV-1/2 test kits using real-time and archived samples of HIV-infected persons from Cameroon. Matched whole blood and OMT specimens were collected prospectively from HIV-positive and HIV-negative persons from different regions of Cameroon and tested using the AWARE BSP and OMT test kits, respectively. These results were compared to the gold standard that included a combination of Determine HIV-1/2 and Enzygnost HIV-1/2. The BSP Rapid test kit was further evaluated using well characterized panels of HIV-2 and HIV-1 group O samples. Cost and end-user analysis of the OMT test kit was done by comparing its actual cost, consumables, safety, bench time and manipulation with other test kits. Of the 732 matched samples, 412 (56.3%) and 320 (43.7%) were from females and males, respectively. Of these samples, 23 (3.1%) gave discordant results between Determine HIV-1/2 and Enzygnost HIV1/2 and were excluded from the analysis. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the AWARE BSP were 100%. The AWARE OMT had 98.8% sensitivity, 98.9% specificity, 98.0% PPV and 99.4% NPV. The results of a well-characterized archived panel of HIV-2 (n=7) and HIV-1 group O (n=3) samples using the AWARE BSP Rapid test kit gave 100% concordance. Total per patient cost of the AWARE OMT rapid test kit was US dollars 4.72 compared to a mean cost of US dollars 7.33 +/- 0.11 for the other test kits. Both the AWARE BSP and OMT Rapid test kits demonstrated high sensitivities and specificities on all samples tested and were well adapted for use in resource-constrained settings with high HIV heterogeneity such as Cameroon. The AWARE HIV-1/2 OMT Rapid test kit appears to be the cheapest, safest and easiest to use compared with other available test kits. PMID- 19543531 TI - Tissue-specific B-cell dysfunction and generalized memory B-cell loss during acute SIV infection. AB - BACKGROUND: Primary HIV-infected patients display severe and irreversible damage to different blood B-cell subsets which is not restored by highly efficient anti retroviral therapy (HAART). Because longitudinal investigations of primary HIV infection is limited by the availability of lymphoid organs, we studied the tissue-specific B-cell dysfunctions in acutely simian immunodeficiency virus (SIV) mac251-infected Cynomolgus macaques. METHODS AND FINDINGS: Experiments were performed on three groups of macaques infected for 14, 21 or 28 days and on three groups of animals treated with HAART for two-weeks either initiated at 4 h, 7 or 14 days post-infection (p.i.). We have simultaneously compared changes in B-cell phenotypes and functions and tissue organization of B-cell areas in various lymphoid organs. We showed that SIV induced a steady decline in SIgG-expressing memory (SIgD(-)CD27(+)) B-cells in spleen and lymph nodes during the first 4 weeks of infection, concomitant to selective homing/sequestration of B-cells to the small intestine and spleen. SIV non-specific Ig production was transiently increased before D14p.i., whereas SIV-specific Ig production was only detectable after D14p.i., coinciding with the presence of CD8(+) T-cells and IgG-expressing plasma cells within germinal centres. Transient B-cell apoptosis on D14p.i. and commitment to terminal differentiation contributed to memory B-cell loss. HAART abrogated B-cell apoptosis, homing to the small intestine and SIV-specific Ig production but had minimal effect on early Ig production, increased B-cell proportions in spleen and loss of memory B-cells. Therefore, virus-B-cell interactions and SIV-induced inflammatory cytokines may differently contribute to early B-cell dysfunction and impaired SIV/HIV-specific antibody response. CONCLUSIONS: These data establish tissue-specific impairments in B-cell trafficking and functions and a generalized and steady memory B-cell loss in secondary lymphoid organs. Characterization of underlying mechanisms would be helpful in designing new therapeutic strategies to dampen B-cell activation and increases HIV/SIV specific antibody response. PMID- 19543532 TI - Ligation of alpha-dystroglycan on podocytes induces intracellular signaling: a new mechanism for podocyte effacement? AB - BACKGROUND: Alpha-dystroglycan is a negatively charged glycoprotein that covers the apical and basolateral membrane of the podocyte. Its transmembrane binding to the cytoskeleton is regulated via tyrosine phosphorylation (pY892) of beta dystroglycan. At the basolateral side alpha-dystroglycan binds the glomerular basement membrane. At the apical membrane, it plays a role in the maintenance of the filtration slit. In this study, we evaluated whether ligation of alpha dystroglycan with specific antibodies or natural ligands induces intracellular signaling, and whether there is an effect on podocyte architecture. METHODOLOGY/PRINCIPAL FINDINGS: Conditionally immortalized podocytes were exposed in vitro to antibodies to alpha-dystroglycan, and to fibronectin, biglycan, laminin and agrin. Intracellular calcium fluxes, phosphorylation of beta dystroglycan and podocyte architecture were studied. Antibodies to alpha dystroglycan could specifically induce calcium signaling. Fibronectin also induced calcium signaling, and led to dephosphorylation of pY892 in beta dystroglycan. Ligation of alpha-dystroglycan resulted in an altered actin architecture, a decreased number of podocyte pedicles and a more flattened appearance of the podocyte. CONCLUSIONS/SIGNIFICANCE: We conclude that ligation of alpha-dystroglycan on podocytes induces intracellular calcium signaling, which leads to an altered cytoskeleton architecture akin to the situation of foot process effacement. In particular the ability of fibronectin to induce intracellular signaling events is of interest, since the expression and excretion of this protein is upregulated in several proteinuric diseases. Therefore, fibronectin-induced signaling via dystroglycan may be a novel mechanism for foot process effacement in proteinuric diseases. PMID- 19543534 TI - Pregnancy outcomes among HIV-infected women undergoing antiretroviral therapy. AB - BACKGROUND: The use of antiretroviral drugs (ARV) to prevent mother-to-child HIV transmission (PMTCT) promises to be effective. However, limited data on the adverse effects of ARV among pregnant women and pregnancy outcomes have been reported in clinical practice. OBJECTIVES: This study aimed to assess adverse effects and outcomes among pregnant HIV-infected women receiving antiretroviral drugs for either antiretroviral therapy (ART) or PMTCT. STUDY DESIGN: This cohort study was at Chonburi Hospital, Thailand, in 2002-2006. RESULTS: A total of 246 pregnant HIV-infected women with the median age (range) of 27 (16-41) years were included in this study. ART was initiated in 16.3% for treatment during ANC, 66.7% for PMTCT during ANC, and 17.1% for PMTCT in labor. Adverse effects, especially anemia, were significantly associated with continuing combined ART in pregnancy (p<0.001). 88.9% delivered normal-term neonates. The prevalence of pre term delivery was 10.2%. Overall, 24 adverse events from 21 pregnant women (8.5%) were noted. A significantly higher prevalence of pre-term delivery was noted in the groups continuing combined ART, or initiating of PMTCT during labor rather than ANC (p=0.02). The incidence of low Apgar scores was 3.6%, and these were associated with initiation of PMTCT during labor (p=0.004). CONCLUSION: Adverse ARV events were more numerous among the pregnant women who needed ART than PMTCT. ANC is beneficial and strongly recommended for all pregnant HIV-infected women for better pregnancy outcomes. PMID- 19543535 TI - Neurobiology of pain in children: an overview. AB - The evaluation of pain in the newborn and the infant is difficult because pain is mainly a subjective phenomenon. Until a few years ago, several myths persisted. First, the myth that children, especially infants, do not feel pain the way adults do, therefore there is no untoward consequences for them. Second, lack of assessment and reassessment for the presence of pain. Third, misunderstanding of how to conceptualise and quantify a subjective experience. Fourth, lack of knowledge of pain treatment. Fifth, the notion that addressing pain in children takes too much time and effort, in ultimate analysis resulting in wasting time. Sixth, fears of hidden -and not easy to diagnose or prevent- adverse effects of analgesic medications, including respiratory depression and addiction. Finally, from a conceptual point of view, high thresholds of pain in neonates and infants were considered to be present by natural character, and useful in protecting infant from pain during birth and transit through the narrow vaginal channel.The present review is focused on the description of different theories on the pain pathogenesis in children. PMID- 19543537 TI - [Not Available]. AB - Optimal design problem are widely known by their multiple performance measures that are often competing with each other. In this paper, an optimal multiproduct batch chemical plant design is presented. The design is firstly formulated as a multiobjective optimization problem, to be solved using the well suited non dominating sorting genetic algorithm (NSGA-II). The NSGA-II have capability to achieve fine tuning of variables in determining a set of non dominating solutions distributed along the Pareto front in a single run of the algorithm. The NSGA-II ability to identify a set of optimal solutions provides the decision-maker DM with a complete picture of the optimal solution space to gain better and appropriate choices. Then an outranking with PROMETHEE II helps the decision maker to finalize the selection of a best compromise. The effectiveness of NSGA II method with multiojective optimization problem is illustrated through two carefully referenced examples. PMID- 19543536 TI - Discrepancy among observational studies: example of naproxen-associated adverse events. AB - BACKGROUND: Observational studies assessing the cardiovascular adverse effect of naproxen have had conflicting results. It is not clear whether variation in population characteristics between studies may explain some of this discrepancy. OBJECTIVE: To determine whether changes in patient characteristics of naproxen users occurred between 1999 and 2004 in Quebec, Canada and to examine whether these temporal changes were accompanied by changes in estimates of naproxen related hospitalizations for gastrointestinal (GI) ulcers and myocardial infarction, using provincial health services administrative databases. METHODS: Demographic, pharmaceutical and physician billing records of patients 65 years and older, who received naproxen or acetaminophen prescriptions between 1999 and 2004 were used. Two identical cohort studies, labeled Study 1 and Study 2 were conducted and their results were compared. One study was confined to the time period 1999-2001 and the other to 2002-2004. Patient characteristics at index date (the date of the first naproxen or acetaminophen prescription during the corresponding period) were compared between the study cohorts in naproxen and acetaminophen users, respectively, and within each study cohort between naproxen and acetaminophen users, using logistic regression models. Cox regression models with time dependent exposure were used to assess the association between naproxen vs acetaminophen and hospitalizations for GI events or AMI, respectively within each study. Results were then compared between the two studies. RESULTS: Study 1 (1999-2001) cohort included 240,568 patients (205,238 acetaminophen and 35,330 naproxen) and Study 2 (2002-2004) cohort included 213,802 patients (193,918 acetaminophen and 19,884 naproxen). Patient characteristics of naproxen and acetaminophen users differed between the two studies. Naproxen users in Study 2 vs Study 1 were slightly younger, less likely to be females, less likely to have concomitant GI disease, less likely to have osteoarthritis and other co morbidities and more likely to have used proton pump inhibitors, antihypertensive agents, anticoagulants, clopidogrel and aspirin. In general, similar changes in patient characteristics were observed in acetaminophen users between the two study cohorts. Compared to acetaminophen (without aspirin), the estimates of the GI risks with naproxen whether, used with or without aspirin, were significantly higher in Study 2 vs Study 1 [Hazard Ratio (HR) (95% CI): 4.94 (3.48, 7.02)] vs [2.22 (1.62, 3.06)], naproxen with aspirin [4.94 (2.93, 8.33) vs 2.47 (1.48, 4.12)], and acetaminophen and aspirin: [2.31 (1.89, 2.82) vs 1.46 (1.20, 1.77)]. The estimate of the AMI risk with naproxen also seemed to be higher in Study 2 vs Study 1, however the increase was not statistically significant [HR (95% CI) in the naproxen group: 1.18 (0.83, 1.67) in Study 1 vs 0.94 (0.70, 1.25) in Study 2], naproxen with aspirin. [1.44 (0.95, 2.18) vs 1.05 (0.68, 1.61)]; and acetaminophen and aspirin. 1.15 (1.01, 1.30) vs 1.10 (0.97, 1.26). CONCLUSION: Variation in patient characteristics in naproxen users was observed between 1999 and 2004. This variation was likely to be accompanied by a variation in patient pre-disposition to GI events that may explain the increase in estimates of naproxen-related GI adverse events observed during this period. PMID- 19543538 TI - Over the Counter Availability of Antituberculosis Drugs in Tbilisi, Georgia in the Setting of a High Prevalence of MDR-TB AB - Georgia, a country of 4.5 million people, has a high incidence of tuberculosis (TB) including drug resistant cases. Easy access and inappropriate use of anti-TB drugs are risk factors for further development of multidrug resistant (MDR)-TB. We carried out an investigation to assess the availability of over the counter anti-TB agents in pharmacies in Tbilisi. During February 2006, 15 pharmacies were randomly selected and the pharmacist at each store was interviewed. We found that all anti-TB medications stocked by these pharmacies were available and sold without a prescription. All 15 pharmacies sold isoniazid, rifampicin, and streptomycin; 13 (87%) of 15 pharmacies also sold pyrazinamide, ethambutol. Second line anti-TB drugs such as amikacin and kanamycin (injectable agents) and older fluoroquinolones (ofloxacin and ciprofloxacin) were available at 13 pharmacies while newer generation fluoroquinolones were less available(3 sold leovofloxacin, none sold moxifloxacin). The ease access and availability of anti TB agents is of a great concern given the high prevalence of TB including MDR-TB in Georgia. The potential for misuse of these anti-TB drugs can lead to the development of further drug resistance. These drugs should only be available by prescription in order to reduce the chance of amplifying drug resistance. PMID- 19543539 TI - Metabolic Demand Stimulates CREB Signaling in the Limbic Cortex: Implication for the Induction of Hippocampal Synaptic Plasticity by Intrinsic Stimulus for Survival. AB - Caloric restriction by fasting has been implicated to facilitate synaptic plasticity and promote contextual learning. However, cellular and molecular mechanisms underlying the effect of fasting on memory consolidation are not completely understood. We hypothesized that fasting-induced enhancement of synaptic plasticity was mediated by the increased signaling mediated by CREB (cAMP response element binding protein), an important nuclear protein and the transcription factor that is involved in the consolidation of memories in the hippocampus. In the in vivo rat model of 18 h fasting, the expression of phosphorylated CREB (pCREB) was examined using anti-phospho-CREB (Ser133) in cardially-perfused and cryo-sectioned rat brain specimens. When compared with control animals, the hippocampus exhibited up to a twofold of increase in pCREB expression in fasted animals. The piriform cortex, the entorhinal cortex, and the cortico-amygdala transitional zone also significantly increased immunoreactivities to pCREB. In contrast, the amygdala did not show any change in the magnitude of pCREB expression in response to fasting. The arcuate nucleus in the medial hypothalamus, which was previously reported to up-regulate CREB phosphorylation during fasting of up to 48 h, was also strongly immunoreactive and provided a positive control in the present study. Our findings demonstrate a metabolic demand not only stimulates cAMP-dependent signaling cascades in the hypothalamus, but also signals to various limbic brain regions including the hippocampus by activating the CREB signaling mechanism. The hippocampus is a primary brain structure for learning and memory. It receives hypothalamic and arcuate projections directly from the fornix. The hippocampus is also situated centrally for functional interactions with other limbic cortexes by establishing reciprocal synaptic connections. We suggest that hippocampal neurons and those in the surrounding limbic cortexes are intimately involved in the metabolism dependent plasticity, which may be essential and necessary for successful achievement of adaptive appetitive behavior. PMID- 19543540 TI - Neurons in the white matter of the adult human neocortex. AB - The white matter (WM) of the adult human neocortex contains the so-called "interstitial neurons". They are most numerous in the superficial WM underlying the cortical gyri, and decrease in density toward the deep WM. They are morphologically heterogeneous. A subgroup of interstitial neurons display pyramidal-cell like morphologies, characterized by a polarized dendritic tree with a dominant apical dendrite, and covered with a variable number of dendritic spines. In addition, a large contingent of interstitial neurons can be classified as interneurons based on their neurochemical profile as well as on morphological criteria. WM- interneurons have multipolar or bipolar shapes and express GABA and a variety of other neuronal markers, such as calbindin and calretinin, the extracellular matrix protein reelin, or neuropeptide Y, somatostatin, and nitric oxide synthase. The heterogeneity of interstitial neurons may be relevant for the pathogenesis of Alzheimer disease and schizophrenia. Interstitial neurons are most prominent in human brain, and only rudimentary in the brain of non-primate mammals. These evolutionary differences have precluded adequate experimental work on this cell population, which is usually considered as a relict of the subplate, a transient compartment proper of development and without a known function in the adult brain. The primate-specific prominence of the subplate in late fetal stages points to an important role in the establishment of interstitial neurons. Neurons in the adult WM may be actively involved in coordinating inter-areal connectivity and regulation of blood flow. Further studies in primates will be needed to elucidate the developmental history, adult components and activities of this large neuronal system. PMID- 19543541 TI - Poly(3,4-ethylenedioxythiophene) as a Micro-Neural Interface Material for Electrostimulation. AB - Chronic microstimulation-based devices are being investigated to treat conditions such as blindness, deafness, pain, paralysis, and epilepsy. Small-area electrodes are desired to achieve high selectivity. However, a major trade-off with electrode miniaturization is an increase in impedance and charge density requirements. Thus, the development of novel materials with lower interfacial impedance and enhanced charge storage capacity is essential for the development of micro-neural interface-based neuroprostheses. In this report, we study the use of conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) as a neural interface material for microstimulation of small-area iridium electrodes on silicon-substrate arrays. Characterized by electrochemical impedance spectroscopy, electrodeposition of PEDOT results in lower interfacial impedance at physiologically relevant frequencies, with the 1 kHz impedance magnitude being 23.3 +/- 0.7 kOmega, compared to 113.6 +/- 3.5 kOmega for iridium oxide (IrOx) on 177 mum(2) sites. Further, PEDOT exhibits enhanced charge storage capacity at 75.6 +/- 5.4 mC/cm(2) compared to 28.8 +/- 0.3 mC/cm(2) for IrOx, characterized by cyclic voltammetry (50 mV/s). These improvements at the electrode interface were corroborated by observation of the voltage excursions that result from constant current pulsing. The PEDOT coatings provide both a lower amplitude voltage and a more ohmic representation of the applied current compared to IrOx. During repetitive pulsing, PEDOT-coated electrodes show stable performance and little change in electrical properties, even at relatively high current densities which cause IrOx instability. These findings support the potential of PEDOT coatings as a micro-neural interface material for electrostimulation. PMID- 19543542 TI - [Not Available]. AB - Since the introduction of nickel-titanium alloy to endodontics, there have been many changes in instrument design, but no significant improvements in the raw material properties, or enhancements in the manufacturing process. Recently, a new method to produce nickel-titanium rotary (NTR) instruments has been developed, in an attempt to obtain instruments that are more flexible and resistant to fatigue. NTR instruments produced using the process of twisting (TF, SybronEndo, Orange, CA) were compared to NTR instruments from different manufacturers produced by a traditional grinding process. The aim of the study was to investigate whether cyclic fatigue resistance is increased for TF NTR files. Tests were performed with a cyclic fatigue device that evaluated cycles to failure of rotary instruments inside curved artificial canals. Results indicated that size 06-25 TF instruments showed a significant increase (P< .05). In the mean number of cycles to failurewhen compared to the other tested 06-25 NTR. Hence, it can be concluded that size 06-25 TF NTR instruments were found to be significantly more resistant to fatigue than those produced with the traditional grinding process. PMID- 19543544 TI - [Not Available]. AB - The Authors describe two cases of alleged malpractice due to overfilling. The aim of this article is to underline some medico-legal aspects regarding the quantity of extruded material which may be considered acceptable and the consequent damage to the patient. Two cases are presented here: In the first case, the dentist's liability is clear due to excessive extrusion of endodontic material beyond the apical region combined with incomplete obturation of the canals. In the second case however, because two different dentists were involved, establishing the connection of causality between their work and the damage reported by the patient was not easy. This situation makes it difficult to establish the limits of potential responsibility, coupled with the complete absence of radiographic signs of periapical rarefaction and the small quantities of material beyond the apex. From a medico-legal point of view, a dentist may be held responsible for compensation and financial expenses of a patient for restoration of damage resulting from a dental procedure.Italian guidelines offer no indications as to when overfilling should be considered the result of a procedural error, or if it fits within the range labelled as "acceptable" and this gap offers extremely subjective interpretations of legal consultants. So, it would therefore be useful to adopt more precise qualitative/dimensional parameters, keeping in mind that the guidelines offer therapeutic recommendations and are not rigid protocols. PMID- 19543543 TI - [Not Available]. AB - Recent advancement in mass spectrometry leads us to a new era of proteomic analysis. Human saliva can be easily collected; however, the complexity of the salivary proteome in the past prevented the use of saliva for proteomic analysis. Here we review the development of proteomic analyses for human saliva and focus on the use of a new mass spectrometric technology known as surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF). SELDI-TOF, a modification of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF), combines the precision of mass spectrometry and the high-through-put nature of protein arrays known as Protein Chips. This technology shows a promising future for salivary proteomic analysis in monitoring treatments and diseases, as well as novel biomarker discovery. PMID- 19543545 TI - [Not Available]. AB - The objective was to assess the effect of examination rates on national caries indices of 5, 12 and 17-year-old children/adolescents in Finland. The data were gathered from patient records of the Public Dental Service (PDS) units (n=205, 73%) and from a national register. The data included PDS-specific total numbers of examined children/adolescents and means of decayed (d/D) teeth, dmft/DMFT values and proportions of caries-free. For analytical purposes, the PDS-specific mean values weighted by the PDS population were calculated to imply the traditionally calculated figures. New PDS-specific examination-rate-adjusted mean values were calculated by using the predicted outcome values at 100% examination rates. The results showed that low examination rates were associated with slightly poorer oral health. The examination-rate-adjusted mean d/D- values indicated better oral health than the traditional indices. The adjustment slightly worsened oral health in proportions of caries-free, and had almost no effect on dmft/DMFT-value. Overall, the influence was modest. The high proportions of healthy children that are examined (against recommendation) and the relatively small number of those having extensive disease (frequently examined as recommended) probably mask the influence of examination rates on the indices in Finland. We conclude that in international comparisons, traditionally calculated indices seem to be sufficiently valid. PMID- 19543546 TI - Comprehension of Health-related Written Materials by Older Adults. AB - This study examined how Flesch Reading Ease and text cohesion affect older adults' comprehension of common health texts. All older adults benefited when high Flesh Reading Ease was combined with high cohesion. Older adults with small working memories had more difficulty understanding texts high in Flesch Reading Ease. Additionally, older adults with low verbal ability or older than 77 years of age had difficulty understanding texts high in text cohesion but low in Flesch Reading Ease. These results imply that writers must increase Flesch Reading Ease without disrupting text cohesion to ensure comprehension of health-related texts. PMID- 19543547 TI - Factors related to quality of life in treatment-adherent, successfully treated HIV patients in France. AB - The objectives of this study were to document the psychosocial characteristics of treatment-adherent, successfully treated HIV patients and to examine the relationships between psychosocial variables. The sample was composed of 133 persons living with HIV, with optimal adherence to antiretroviral therapy and with no detectable viral load. The psychosocial profile of the cohort showed that participants had moderate levels of stress, used a wide range of strategies to adjust to their situation and perceived their social support to be good. As well, they evaluated their quality of life (Medical Outcome Study-HIV) as moderate. Quality of life in psychological domain is largely explained by perceived stress (53%). Quality of life in physical domain is largely dependent on discomfort reported arising from HIV-associated symptoms and treatment (39%). Findings suggest that treatment-adherent, successfully treated patients with HIV are still a fragile population, and that it is essential to provide interventions that reinforce their ways of coping. PMID- 19543548 TI - [Not Available]. AB - INTRODUCTION: E-Learning methods such as webcasting are being used increasingly in healthcare education, including that of nurses and midwives. Webcasting means live synchronous broadcasting over the internet, where students participate simultaneously in text 'chat room' interactive discussions when logged on to a webpage where they can see and hear a presentation such as a PowerPoint lecture, a list of other participants, and access 'chat rooms'. AIMS: This paper reports student participation and satisfaction with the use of webcasting in a third year undergraduate nursing and midwifery research methods module in one higher education institution faculty of health and social work in the southwest of England, with students from distributed geographical locations. MATERIALS AND METHODS: Students chose either webcasts or face-to-face lectures. Following each of the four webcasts, a web-based evaluation questionnaire was administered in a cross-sectional survey design. RESULTS: Two thirds of students took part in webcasts and found them to be an acceptable teaching and learning strategy. Travel and cost savings were noted through not travelling to the main university campus, and these were statistically significantly correlated with students' perception of gaining from the module and their overall satisfaction with webcasting. Across the four webcasts 5446 purposeful messages were posted indicating engagement with the material under study. CONCLUSIONS AND RECOMMENDATIONS: Webcasting is an effective teaching and learning strategy which is popular with students, allows remote access to teaching and learning, and offers time and cost savings to students. Further research is required to investigate the educational potential of this new technology. PMID- 19543549 TI - [Not Available]. AB - Human saliva can be separated by centrifugation into cell pellet and cell-free supernatant, which are called cellular phase and liquid phase in this study. While it is well documented that the cellular phase of saliva contains hundreds of oral bacteria species, little is known whether the liquid phase of saliva contains any information related to oral microbiota. In this study, we analyzed the bacterial nucleic acid contents of the liquid phase of saliva. Using primers universal to most eubacterial 16S rDNA, we detected large amounts of bacterial 16S rRNA and rDNA in the cell-free phase of saliva. Random sequencing analysis of forty PCR amplicons from the cell-free phase of saliva led to 15 operational taxonomic unit (OTU) groups. Furthermore, using denaturing gradient gel electrophoresis (DGGE), we compared 16S rRNA/rDNA profiles derived from liquid phases and cellular phases of saliva samples, and found positive correlations (Pearson Correlation=0.822, P<0.001) between these sample groups. These findings indicate that the liquid phase of saliva contains numerous bacterial 16S rRNA/rDNA molecules that have correlations with bacteria existing in the cellular phase. PMID- 19543551 TI - [Not Available]. AB - Relatively little is known about the microbial communities adapted to soil environments contaminated with aged complex hydrocarbon mixtures, especially in the subsurface soil layers. In this work we studied the microbial communities in two different soil profiles down to the depth of 7 m which originated from a 30 year-old site contaminated with petroleum hydrocarbons (PHCs) and from a clean site next to the contaminated site. The concentration of oxygen in the contaminated soil profile was strongly reduced in soil layers below 1 m depth but not in the clean soil profile. Total microbial biomass and community composition was analyzed by phospholipid fatty acid (PLFA) measurements. The diversity of fungi and actinobacteria was investigated more in detail by construction of rDNA based clone libraries. The results revealed that there was a significant and diverse microbial community in subsoils at depth below 2 m, also in conditions where oxygen was limiting. The diversity of actinobacteria was different in the two soil profiles; the contaminated soil profile was dominated by Mycobacterium related sequences whereas sequences from the clean soil samples were related to other, generally uncultured organisms, some of which may represent two new subclasses of actinobacteria. One dominating fungal sequence which matched with the ascomycotes Acremonium sp. and Paecilomyces sp. was identified both in clean and in contaminated soil profiles. Thus, although the relative amounts of fungi and actinobacteria in these microbial communities were highest in the upper soil layers, many representatives from these groups were found in hydrocarbon contaminated subsoils even under oxygen limited conditions. PMID- 19543552 TI - [Not Available]. AB - As a coagulase negative Staphylococcus species, S. caprae is not considered as a clinically-significant member, unlike S. epidermidis. In this report, we describe a case of sepsis resulting from S. caprae infection. This relatively young woman was in generally good health and contracted S. caprae most probably during her treatment of an acute pulmonary embolism. The purpose of this report is to raise awareness of this otherwise innocuous staphylococcal species in clinical settings. PMID- 19543550 TI - [Not Available]. AB - BACKGROUND: The healing potential of platelet growth factors has generated interest in using Platelet-Rich Plasma (PRP) in ridge preservation procedures. A canine study was performed to determine if extraction sites treated with platelet rich fibrin matrix (PRFM) exhibit enhanced healing compared to sites treated with non-viable materials. METHODS: Four dog's extraction sockets were treated individually with PRFM, PRFM and membrane, Demineralized Freeze-Dried Bone Allograft (DFDBA) and membrane, PRFM and DFDBA, and untreated control. Treatment sequencing permitted clinical and histologic evaluation of healing at 10 days, 2, 3, 6 and 12 weeks. RESULTS: Healing was more rapid in the PRFM and PRFM and membrane sites. By 3 weeks those sockets had osseous fill. Sites containing DFDBA had little new bone at 6 weeks. By 12 weeks those sockets had osseous fill but DFDBA particles were still noted in coronal areas. CONCLUSIONS: PRFM alone may be the best graft for ridge preservation procedures. ADVANTAGES: faster healing, and elimination of disadvantages involved in using barrier membranes. PMID- 19543553 TI - [Not Available]. AB - Production of IL-12 and IFN-gamma secretion are important components of the protective host response against the intracellular bacterial pathogen, Salmonella typhimurium. While infection with Salmonella does elicit this T helper type 1 response, its magnitude does not appear to be sufficient to prevent infection or limit pathogenesis. Therefore we have investigated factors which might limit a T helper type 1 response following infection. Previously we found that infection of antigen presenting cells with Salmonella dramatically increases cyclooxygenase-2 (COX-2) activity, resulting in high levels of prostaglandin E2 (PGE2). Since PGE2 production can have profound effects on initiation of T helper type 1 responses, we questioned whether this mediator might limit antigen-specific T cell activation. Here we show that blockage of COX-2 activity with the selective inhibitor celecoxib leads to enhancement of the T helper type 1 components stimulated by Salmonella infection. In vitro studies demonstrate the induction of IL-12 and IFN-gamma upon Salmonella exposure, which are further increased following COX-2 inhibition. Taken together these in vitro studies suggest that COX-2 activity can limit a salmonella-initiated T helper type 1 response. PMID- 19543554 TI - Chemerin and the recruitment of NK cells to diseased skin. AB - Natural killer (NK) cells play a major role in the initial control of many viral pathogens and in the rejection of tumors. Consistent with their roles as immune sentinels, NK cells are found in inflamed skin, including lichen planus, psoriasis and atopic dermatitis (AD) lesions. In oral lichen planus lesions, the recruitment as well as intradermal colocalization of NK cells and pDC (plasmacytoid dendritic cells) appear to be mediated by chemerin, a recently identified protein ligand for chemokine-like receptor 1 (CMKLR1), a chemoattractant receptor expressed by both cell types. Dendritic cells can regulate NK cell activity, and NK cells can regulate DC-mediated responses. Since chemerin was recently implicated in recruitment of pDC to psoriatic skin, in this work we determined whether chemerin facilitates interactions between NK and pDC in psoriatic plaques through controlling influx of NK cells to diseased skin. We demonstrate that circulating NK cells from normal donors as well as psoriasis and AD patients respond similarly in functional migration assays to chemerin. However, differences in the distribution of NK cells and pDC in skin lesions suggest that recruitment of both NK cells and pDC is unlikely to be controlled solely by chemerin. PMID- 19543555 TI - Extradermal melanin transfer? Lack of macroscopic spleen melanization in old C57BL/6 mice with de-synchronized hair cycle. AB - In quest of alternate, extradermal path of melanin transfer from skin to the visceral organs, we suggested that some portions of such melanin may be deposited in the spleen, which in young black C57BL/6 mice is often melanized. Here, we confirm these observation using young C57BL/6 female mice (up to 17 weeks) and show that this phenomenon cannot be observed in old animals where the hair cycle is not synchronized any more. The experiments were carried out both on spontaneous and depilation-induced hair cycle. We have checked it as a side observation over many other experiments carried out on young and old C57BL/6 female mice (up to 2.5 years of life). The presence or absence of melanin in the spleens was checked macroscopically, and histologically by Fontana-Masson (FM) staining, and synchronization of the hair cycle - by standard histomorphometric analysis of the back skin hair follicles. In about 40% of old spleens black FM stainable "debris" could be found under closer histological examination. This study shows that, at least in part, the phenomenon of splenic melanosis in C57BL/6 mice can be correlated with the synchronized skin melanization parallel to the hair cycle progress, and that splenic melanin undergoes gradual degradation during the mouse life. PMID- 19543556 TI - Fungi pathogenic to humans: molecular bases of virulence of Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus. AB - The frequency of severe systemic fungal diseases has increased in the last few decades. The clinical use of antibacterial drugs, immunosuppressive agents after organ transplantation, cancer chemotherapy, and advances in surgery are associated with increasing risk of fungal infections. Opportunistic pathogens from the genera Candida and Aspergillus as well as pathogenic fungi from the genus Cryptococcus can invade human organism and may lead to mucosal and skin infections or to deep-seated mycoses of almost all inner organs, especially in immunocompromised patients. Nowadays, there are some effective antifungal agents, but, unfortunately, some of the pathogenic species show increasing resistance. The identification of fungal virulence factors and recognition of mechanisms of pathogenesis may lead to development of new efficient antifungal therapies. This review is focused on major virulence factors of the most common fungal pathogens of humans: Candida albicans, Aspergillus fumigatus and Cryptococcus neoformans. The adherence to host cells and tissues, secretion of hydrolytic enzymes, phenotypic switching and morphological dimorphism contribute to C. albicans virulence. The ability to grow at 37 degrees C, capsule synthesis and melanin formation are important virulence factors of C. neoformans. The putative virulence factors of A. fumigatus include production of pigments, adhesion molecules present on the cell surface and secretion of hydrolytic enzymes and toxins. PMID- 19543557 TI - Exogenous nitric oxide inhibits shedding of ADAM17 substrates. AB - Both ADAM17, the secretase responsible for the shedding of ectodomains of numerous membrane proteins including TNF and its receptors, as well as nitric oxide synthesized by inducible nitric oxide synthase play regulatory roles in inflammation and tumor progression. We analyzed the effect of endogenous and exogenous nitric oxide on the expression and activity of ADAM17 in murine endothelial cells and a monocyte/macrophage cell line. We found that endogenous nitric oxide influenced neither ADAM17 mRNA level nor the shedding of two ADAM17 substrates, TNF and TNFR1. Exogenous NO significantly diminished the release of TNF and TNFR1 without affecting the ADAM17 transcript level. Our data seem contrary to a previous report that showed the activation of ADAM17 by nitric oxide (Zhang et al., 2000, J Biol Chem 275: 15839-15844). We discuss potential mechanisms of NO-mediated inhibition of ectodomain shedding and possible reasons of discrepancy between our results and the previous report. PMID- 19543559 TI - Lansoprazole is an uncompetitive inhibitor of tissue-nonspecific alkaline phosphatase. AB - Lansoprazole, a known H(+)/K(+)-ATPase inhibitor, is currently used as a therapeutical option for the initial treatment of gastroesophageal reflux disease. Recently, lansoprazole has been found to be an inhibitor of cytosolic PHOSPHO1 (a phosphatase which hydrolyses phosphocholine and phosphoethanolamine), providing a possible therapeutical target to cure pathological mineralization. Since PHOSPHO1 is present inside matrix vesicles, we tested the effect of lansoprazole on matrix vesicles containing several key enzymes for the mineralization process including tissue-nonspecific alkaline phosphatase. We found that lansoprazole can inhibit in an uncompetitive manner tissue-nonspecific alkaline phosphatase. A K(i) value of 1.74 +/- 0.12 mM has been determined for the inhibition of tissue-nonspecific alkaline phosphatase by lansoprazole. Lansoprazole, currently used for treating gastroesophageal disease, by inhibiting PHOSPHO1 and tissue-nonspecific alkaline phosphatase could prevent hydroxyapatite deposition disease and could serve as an adjunct treatment for osteoarthritis. PMID- 19543558 TI - Reference genes for gene expression studies on non-small cell lung cancer. AB - STUDY OBJECTIVE: The aim of this study was to test a panel of 6 reference genes in order to identify and validate the most suitable reference genes for expression studies in paired healthy and non-small cell lung cancer tissues. METHOD: Quantitative real-time PCR followed by the NormFinder- and geNorm-based analysis was employed. The study involved 21 non-small cell lung cancer patients. RESULTS: The analysis of experimental data revealed HPRT1 as the most stable gene followed by RPLP0 and ESD. In contrast, GAPDH was found to be the least stable gene. HPRT1 together with ESD was revealed as the pair of genes introducing the least systematic error into data normalization. Validation by bootstrap random sampling technique and by normalizing exemplary gene expression data confirmed the results. CONCLUSION: Although HPRT1 and ESD may by recommended for data normalization in gene expression studies on non-small cell lung cancer, the suitability of selected reference genes must be unconditionally validated prior to each study. PMID- 19543560 TI - [Cocaine-induced agitated delirium, a different consequence of cocaine addiction]. AB - In the past 10 years, chronic drug abuse health problems have become complex medical-legal problems. These include Cocaine-Induced Agitated Delirium, an idiosyncratic illness appears 1-2 hours after regular drug intake and can cause death without overdose is detected at autopsy. This review studies the molecular changes caused by cocaine abuse that derived into Cocaine-Induced Agitated Delirium. The molecular-clinical correlation links the phenomena induced by the abuse expressed at different levels of complexity, from the biochemical to the social. The purpose here is to induce a greater awareness of this illness to improve its prevention, to obtain its early diagnosis and to achieve its appropriate therapeutic-legal approach. Cocaine-Induced Agitated Delirium should be considered as the result of several unseen changes, that if they reach a critical threshold trigger the fatal outcome. This makes the abuse of cocaine and the individual predisposition solely responsible for its appearance. PMID- 19543561 TI - [A neuropsychoanalytic freudian model of psychic trauma and memory. Theoretical and clinical applications]. AB - The traumatic memory is conceptualized by means of an amplified Freudian neuropsychoanalytic model using a contemporary memory system based on its contents, conscious and unconscious recollection (explicit and implicit memories) highlighting the validity of the Freudian discoveries. This is then related to the psychoanalytical theories of consciousness, affects and thinking. Particular importance is given to Freud's seduction theory, its relation to memory and the clinical application of these concepts to the basic organization of the personality, together with the relation to Bowlby's concept of emotional deprivation. The development and working trough of trauma is postulated as a vector to make "real" or phantasized trauma unconscious through repression in neurosis, splitting in borderline personality organization, and primitive mechanisms of projection in psychosis. PMID- 19543562 TI - [Randomized clinical trials and real clinical practice]. AB - One of the emerging problems in modern medicine is that part of its highly efficacious treatments do not show significant effectiveness in real world systems of care. Efficacy studies address the appropriate dosages, short term response and feasibility of treatments in carefully selected populations, but they do not necessarily provide information for decisions in clinical practice. This review aims to present strengths and limitations of different methodological types of trials and to offer an overview of how knowledge from clinical trials can be used for clinical practice. The important effect of funding source on the outcome of randomized controlled trials is discussed. Some key questions in the treatment assessment of depression, schizophrenia and different medical conditions are discussed, with a focus on the possibilities and restrictions of translating clinical trial results into real-world settings. Empirical evidence shows that although randomized controlled trials are the gold standard for proving efficacy of a therapeutic procedure they often suffer from funding source bias and from lack of generalizability. Effectiveness studies evaluate effects of treatments under conditions approximating usual care. Another key area that can be addressed by effectiveness studies is the impact on important health policy measures such as disability days, days of work or medical costs, etc. Conclusions show that the future assessment of treatment regimes for clinical utility requires less biased efficacy studies and more effectiveness studies addressing major issues from all relevant perspectives. PMID- 19543563 TI - [STAR* D study: searching strategies regarding lack of response to an SSRI]. AB - STAR*D, the biggest investigation ever conducted on the treatment of major depression, was aimed to determine prospectively which of several treatments is the most effective "next step" for patients who do not reach remission with an initial or subsequent treatment or who cannot tolerate the treatment. Its main findings show that: 1) Monotherapy remission rates are low; 2) There were no outcome differences in changing to another SSRI, to a dual reuptake inhibitor o to another mechanism of action. 3) Augmentation strategies worked perhaps better in patients with partial response and good citalopram tolerability. There was a slight advantage of bupropion over buspirone. T3 had better tolerability and was easier to use than lithium; 4) There was no difference between cognitive therapy as a switch or as augmentation strategy versus medication as a switch or augmentation strategy. Antidepressants were faster in augmentation. 5) There were no statistical differences in remission rates among any of the medications compared in this study; 6) Remission rates drop level after level, more substantially after two failed treatments. Conversely, recurrence rates rise progressively; 7) Patients who achieve remission are less likely to relapse than patients who have only responded. PMID- 19543564 TI - [Effectiveness trials in chronic schizophrenic patients: CATIE. What can we learn?]. AB - The need for practical clinical trials on schizophrenic patients led to the development of the CATIE to study the effectiveness of antipsychotics in the "real world". This study has different phases that are described in this article, detailing their methodology, results and limitations. There were no significant differences between the antipsychotics of first and second generation. Olanzapine appeared as a drug of higher effectiveness but with important metabolic side effects. Clozapine had higher efficacy in patients that had not responded to an antipsychotic administered in a previous phase of this study. Risperidone appeared to be more convenient for patients that abandoned their antipsychotic treatment because of intolerability. Antipsychotics still leave a lot of therapeutic expectations without fulfillment. Other therapeutic measures should be carried out to accomplish a more comprehensive treatment. The treatment for each patient must be specifically designed and the achievement of compliance must be considered of great importance. PMID- 19543565 TI - [Atypical antipsychotics and CATIE-AD. Is this a turning point?]. AB - The Clinical Antipsychotic Trials of Intervention Effectiveness -Alzheimer Disease (CATIE-AD) is the first 36 weeks effectiveness trial of atypical antipsychotics in patients with Alzheimer's disease presenting with behavioral and psychiatric symptoms of dementia (BPSD). There were no significant differences between atypical antipsychotics and placebo in effectiveness measures which also included a cost-benefit analysis. This article not only reviews this landmark study but also summarizes the complex relationship between antipsychotics and BPSD during the last years. PMID- 19543566 TI - [Revision of systematic treatment enhancement program for bipolar disorder (STEP BD)]. AB - The STEP-BD is a public initiative, created to generate data, obtained in pragmatic or real life research environments, from the bipolar disorder. The primary objectives are: to investigate the effectiveness of therapeutic interventions in bipolar disorder, the impact on the disease course of these interventions in the 'real life' and to raise the knowledge about the disease. This program included in six years, n = 4361 patients. This program is not a study in phases but an infrastructure for more than 40 published studies and 30 conference presentations. These studies aim to capture the heterogeneity of the disease in their different clinical presentations, treatments, comorbidities, functionality and quality of life. Patients entering the program have a common assessment. The program consists of two main pathways: the standard treatment and randomized treatment of acute bipolar depression, bipolar depression and refractory relapse prevention. In addition several studies completed the program in an attempt to give coherence to the different clinical presentations of this disease, different treatments and interventions for similar clinical phenomena. PMID- 19543568 TI - The lost years: the impact of cirrhosis on the history of jazz. PMID- 19543569 TI - Therapy for hepatitis B: 'la nouvelle vague' Interview by Paul C. Adams. PMID- 19543570 TI - A noninvasive imaging technique to evaluate therapeutic efficacy after injection of n-butyl-2- cyanoacrylate tissue adhesive into gastric varices: a case report. AB - A novel use of multidetector computed tomographic intravenous (MDCT IV) portography in the evaluation of gastric varices treated with tissue adhesive is described. A 55-year-old man presented with upper gastrointestinal hemorrhage as a result of bleeding gastric varices. The patient was stabilized and the gastric varices were treated with n-butyl-2-cyanoacrylate (two injections, total 7.5 mL). MDCT IV portography performed after injection revealed thrombosis of all but one of the submucosally based gastric varices. The endoscopist who performed repeat endoscopy three weeks later was then able to direct therapy at the remaining patent submucosally based gastric varix. This represents the first reported use of MDCT IV portography in the evaluation of treatment adequacy in a patient with gastric varices treated with n-butyl-2-cyanoacrylate. PMID- 19543571 TI - Esophageal papillomatosis complicated by squamous cell carcinoma. AB - Esophageal papillomatosis is a very rare condition that is believed to have a benign clinical course. Recent reports underscore the potential development of a malignancy in association with squamous papillomatosis of the esophagus. A case of esophageal papillomatosis complicated by the development of esophageal invasive squamous cell carcinoma diagnosed after esophagectomy, despite multiple nondiagnostic endoscopic biopsies, is described. The patient also developed squamous cell carcinoma in the oral cavity and pyloric channel. The finding of extensive esophageal papillomatosis and unremitting dysphagia symptoms should prompt investigations into an underlying associated malignancy. PMID- 19543572 TI - Improving access to care by allowing self-referral to a hepatitis C clinic. AB - BACKGROUND: Estimates suggest that more than 250,000 Canadians are infected with hepatitis C virus (HCV), but less than 10% have been treated. Access to specialists in Canada is usually via health care professional (HCP) referral and, therefore, may be a barrier to HCV care. However, clinics that operate in conjunction with the Hepatitis Support Program, Edmonton, Alberta, allow self referral. It is hypothesized that this improves access to care without increasing inappropriate referrals. OBJECTIVE: To compare the baseline characteristics and outcomes of HCV patients who self-referred with those who were HCP-referred. METHODS: Data were collected from the Hepatitis Support Program HCV database and chart reviews. RESULTS: Between December 17, 2002, and December 31, 2007, 1563 patients were referred including 336 self- (21.5%) and 1227 HCP-referrals (78.5%). Self- and HCP-referred patients were similar in terms of age (mean [+/- SD] 43.0+/-10.3 years versus 43.9+/-10.0 years, respectively; P=0.18), sex (56.8% versus 62.0% [men], respectively; P=0.08) and risk factors for HCV (P=0.3), with 49.7% and 52.6%, respectively, identifying injection drug use as the primary risk factor. The two groups had similar HCV genotype distributions and liver biopsy fibrosis scores with similar treatment rates (31.3% versus 33.2%; P=0.6). Treatment outcomes were excellent (sustained virological response 40.2% for genotype 1, 67% for genotypes 2 and 3) in patients completing therapy and were similar between the two groups. CONCLUSION: Self-referred patients comprised 21.5% of patients accessing care in the clinic. Self- and HCP-referred patients had similar characteristics, treatment rates and outcomes. Facilitating self referral to an HCV clinic can improve access to care, including risk reduction education and HCV treatment. PMID- 19543573 TI - Liver biopsies for chronic hepatitis C: should nonultrasound-guided biopsies be abandoned? AB - BACKGROUND/OBJECTIVE: Liver biopsy has been the gold standard for grading and staging chronic hepatitis C virus (HCV)- mediated liver injury. Traditionally, this has been performed by trained practitioners using a nonimage-guided percutaneous technique at the bedside. Recent literature suggests an expanding role for radiologists in obtaining biopsies using an ultrasound (US)-guided technique. The present study was undertaken study to determine if the two techniques produced liver biopsy specimens of similar quality and hypothesized that at our institution, non-US-guided percutaneous liver biopsies for HCV would be of higher quality than US-guided specimens. METHODS: Liver biopsies from 100 patients with chronic HCV infection (50 consecutive US-guided and 50 consecutive non-US-guided), were retrospectively identified using a hospital histopathology database. All original biopsy slides were coded and prospectively reanalyzed by a single hepatopathologist who was blinded to the technique used in obtaining the biopsy. Additionally, all liver biopsies for chronic HCV infection completed at the centre from 1998 to 2007 were identified and the technique used was recorded. Biopsy quality was determined primarily by the number of complete portal tracts (CPTs) identifiable in the slides. The total length of specimen and the degree of fragmentation were secondary outcome measures. RESULTS: There was a slight difference observed between the US-guided and non-US-guided groups in mean age (46.3 years versus 42.5 years, respectively; P=0.018) but no differences in sex, presence of cirrhosis, bilirubin, creatinine, international normalized ratio, and grade or stage of disease. Biopsies obtained using the US-guided technique produced higher quality specimens than the non-US-guided technique based on our primary outcome of number of CPTs in the biopsy (11.8 versus 7.4; P<0.001). US guided specimens also were longer (24.4 mm versus 19.7 mm; P=0.001), had less fragmentation (P=0.016), and a higher overall histopathological quality assessment (P=0.026) than the non-US-guided biopsies. However, there was no significant difference between the two groups in the ability to grade and stage the disease (96% US-guided versus 90% in non-US-guided (P=0.20). Over a 10-year period, 763 biopsies for chronic HCV infection were identified with an obvious trend toward the increased use of US-guided technique observed at 2% in 1998 to 85% in 2007. CONCLUSIONS: US-guided liver biopsies for chronic HCV are the most common method of obtaining specimens at the Kingston General Hospital, Kingston, Ontario, and are of higher quality than non-US-guided specimens. However, there is no significant difference in the two techniques in the ability to grade and stage chronic HCV. PMID- 19543574 TI - Bone mineral densities in individuals with Gilbert's syndrome: a cross-sectional, case-control pilot study. AB - BACKGROUND: Unconjugated bilirubin inhibits osteoblastic proliferative activity in vitro, raising the possibility that Gilbert's syndrome (GS) patients are at increased risk of osteoporosis. OBJECTIVES: To compare bone mineral density (BMD), serum parathyroid hormone (PTH), C-telopeptide (CTX) and osteocalcin levels in GS subjects versus matched controls in a cross-sectional, case-control study. METHODS: BMD determinations were obtained with central dual energy x-ray absorptiometry. Serum PTH, CTX and osteocalcin levels were measured by enzyme immunoassay. RESULTS: A total of 17 GS and 30 control subjects were studied. Overall, there were no significant differences in BMD, PTH, CTX or osteocalcin levels between the two groups. However, when older (older than 40 years of age) and younger (40 years of age and younger) cohorts were considered separately, the older GS cohort had significantly decreased total hip BMD, T scores and Z scores, and femoral neck BMD, T scores and Z scores (P<0.005 for each parameter, respectively) compared with older control subjects. Serum osteocalcin levels were lower in the older versus younger GS cohort (P=0.006). An inverse correlation existed between all subjects' serum unconjugated bilirubin levels and total body BMD determinations (r=-0.42; P=0.04). On univariate analysis, the association between serum unconjugated bilirubin and total body BMD was not significant (P=0.066), nor was serum unconjugated bilirubin identified as a risk factor for low BMD when entered into multivariate analyses. CONCLUSIONS: The results of the present pilot study warrant further research involving larger numbers of subjects and longitudinal measurements to determine whether GS is associated with decreased BMD, particularly in older GS subjects. PMID- 19543575 TI - Who should be performing liver biopsies? PMID- 19543576 TI - Relevance of segmental colitis with diverticulosis (SCAD) to other forms of inflammatory bowel disease. AB - A well localized inflammatory process involving only the sigmoid colonic segment associated with diverticulosis (SCAD), has become increasingly recognized as a distinct clinical and pathological disorder, usually described in older adults, often with rectal bleeding. Although some resolve spontaneously, most patients appear to respond to treatment only with 5-aminosalicylate. Endoscopic evaluation reveals a nonspecific inflammatory process localized in the sigmoid colon that usually completely resolves with histologically normal colonic mucosa. Recurrent symptoms with evidence of recurrent segmental colitis may occur, but most have an entirely benign clinical course. Further definition of the underlying molecular signalling that occurs in this apparently distinctive disorder may be critically important to understand the elements of a colonic inflammatory process that can completely and spontaneously resolve. PMID- 19543577 TI - Abnormal hematological indices in cirrhosis. AB - Abnormalities in hematological indices are frequently encountered in cirrhosis. Multiple causes contribute to the occurrence of hematological abnormalities. Recent studies suggest that the presence of hematological cytopenias is associated with a poor prognosis in cirrhosis. The present article reviews the pathogenesis, incidence, prevalence, clinical significance and treatment of abnormal hematological indices in cirrhosis. PMID- 19543597 TI - Photophysical and magnetic properties of magnetic silica gel-supported silicon phthalocyanine complexes. AB - Enhanced S(1)--> T(1) intersystem crossing of silicon phthalocyanine (SiPc) owing to iron oxide nanoparticles and intense MCD signal of diamagnetic SiPc have been experimentally shown by using SiPcs supported on magnetic silica gel, in which the iron oxide nanoparticles were effectively dispersed. PMID- 19543598 TI - UV SERS at well ordered Pd sphere segment void (SSV) nanostructures. AB - Ultraviolet laser excited surface-enhanced Raman scattering was obtained for the first time at the well ordered palladium sphere segment void (SSV) nanostructures, using adenine as the probe molecule, and the UV-SERS enhancement is found to be correlated well with the plasmon absorption of Pd SSVs in the UV region. PMID- 19543599 TI - Maximum work in minimum time from a conservative quantum system. AB - This paper considers the problem of obtaining maximum work from a conservative quantum system corresponding to a given change in an external parameter in the Hamiltonian. The example we present is a non-interacting collection of harmonic oscillators with a shared frequency omega which changes from a given initial to a given final value. The example is interesting for its role in experiments at ultra-low temperatures and for probing finite-time versions of the third law of thermodynamics. It is also the simplest system displaying quantum friction, which represents loss mechanisms in any reversible prelude to a thermal process. The example leads to a new type of availability. It is also the first example of a minimum time for transitions between thermal states of a thermodynamic system. PMID- 19543600 TI - Quantum chemical studies of the adsorption of single acetone molecules on hexagonal ice I(h) and cubic ice I(c). AB - The interaction energies of free acetone molecules with surfaces of two different ice polymorphs have been investigated by quantum chemical methods. Special emphasis has been given to sites for adsorption on the (0001) surface of hexagonal ice (I(h)) and the (1[combining macron]01) surface of cubic ice (I(c)), respectively. The structural optimisations made use of conventional electronic structure methods including HF and B3LYP using moderate basis sets up to 6 31+G(d) as well as local and ONIOM methods using 2 or 3 layers which were treated at different levels of theory. The adsorption energies at T = 0 K were calculated for the optimised adsorption geometries performing single points at the B3LYP, MP2 and LMP2 level in conjunction with valence triple-zeta basis sets up to 6 311+G(d,p). Including corrections for basis set superposition errors (BSSE) the most extensive calculations provide adsorption energies (T = 0 K) of -39.1 and 57.5 kJ mol(-1) for the energetically most favourable sites for adsorption of a single acetone molecule on ice I(h) and ice I(c), respectively. By vibrational analysis this can be transformed to adsorption enthalpies at around a temperature of 200 K yielding values of -31.5 for adsorption on ice I(h) and -49.9 kJ mol(-1) for adsorption on ice I(c). The current results support experimental observations of Behr et al. (J. Phys. Chem. A, 2006, 110, 8098) in which evidence was presented that acetone adsorbs on ice around 200 K at two different sites; each of which has a different adsorption enthalpy. PMID- 19543601 TI - Experimental and theoretical studies of complexes of [Pb(m)Ag](-) (m = 1-4). AB - The metal clusters [Pb(m)Ag](-) (m = 1-4) are studied by photoelectron (PE) spectra and density functional theory (DFT). The adiabatic electron affinity (EA) and vertical detachment energy (VDE) of [Pb(m)Ag](-) are obtained from PE spectra at 308 nm. Theoretical calculation is carried out to search for the lowest-energy geometry and elucidate their structures and bonding mode. By comparing the theoretical results, including EA, VDE and simulated density of state (DOS) spectra, with the experimental determination, the lowest-energy structures for each species are obtained. The analysis of the molecular orbital composition provides evidence that the silver atom binds on lead clusters through an Ag-Pb sigma bond. Moreover, the clusters of [Pb(3)](2-), [Pb(4)](2+), Pb(4) and [Pb(4)](2-) are investigated for aromaticity. PMID- 19543602 TI - Hydrides and dimers of C(58) fullerenes: structures and stabilities. AB - A density functional theory (DFT) study of fullerene hydrides C(58)H(2x) (2x = 2,4,...,34) is presented. We consider two relevant isomers, the most stable classical isomer C(58)-C(3v) and the energetically close non-classical isomer C(58)-C(s), which contains a heptagonal ring. Iterative pairwise addition of hydrogen atoms to only the energetically favoured products of the previous iteration yields a set of low energy structures for each composition. From these, low energy pathways are extracted. Analysis of the C-H binding energies along the reaction pathways is performed to identify particularly stable hydride compositions. These are 2x = 6,18,28,34 for C(58)-C(s) and 2x = 10,26,30 for C(58)-C(3v). We therefore suggest that these hydrides are preferably formed in hydrogenation experiments and that it should be possible to distinguish between the two C(58) isomers. We further investigate the dimer formation based on low energy C(58)H(2) addition patterns. All dimers show binding energies of more than 1 eV whereby dispersion interactions play a significant role. Both C(58) isomers can also undergo further aggregation. This leads us to the conclusion that in the absence of other reactant molecules C(58) will form intercage bonds and cannot be isolated in molecular form, which is in accordance with experimental results. PMID- 19543603 TI - Onset of carbon-carbon bonding in the Nb(5)C(y) (y = 0-6) clusters: a threshold photo-ionisation and density functional theory study. AB - We have used photo-ionisation efficiency spectroscopy to determine the ionisation potentials (IPs) of the niobium-carbide clusters, Nb(5)C(y) (y = 0-6). Of these clusters Nb(5)C(2) and Nb(5)C(3) exhibit the lowest IPs. Complementary density functional theory calculations have been performed to locate the lowest energy isomers for each cluster. By comparing the experimental IPs with those calculated for candidate isomers, the structures of the Nb(5)C(y) clusters observed in the experiment are inferred. For all these structures, the underlying Nb(5) cluster has either a "prolate" or "oblate" trigonal bipyramid geometry. Both Nb(5)C(5) and Nb(5)C(6) are shown to contain carbon-carbon bonding in the form of one and two molecular C(2) units, respectively. PMID- 19543604 TI - Heterogeneous chemistry of toluene, kerosene and diesel soots. AB - Soot samples as potential mimics of atmospheric aerosols have been produced from the combustion of toluene, kerosene and diesel in order to compare the nature of soot produced from a simpler material, toluene, with soots from the fuels kerosene and diesel. Characterisation of the soots using elemental analysis, infrared spectroscopy, solvent extraction, thermal desorption and electron microscopy techniques before and after reaction with ozone allows assessment of the reactivity of soots from these different fuels. Despite the production of toluene and kerosene soots from identical combustion conditions, strong differences in structure and reactivity are observed in terms of their reaction with ozone. However, toluene soot is a much better mimic of diesel soot. It is proposed that the differing reactivities of the soots is related to the nature of the organic carbon and structure of the elemental carbon which vary with soots from the different fuels. PMID- 19543605 TI - Self-assembled dithiothreitol on Au surfaces for biological applications: phospholipid bilayer formation. AB - Self-assembly of dithiothreitol (DTT) on Au(111) from solution deposition has been studied by X-ray photoelectron spectroscopy and electrochemical data. DTT molecules self-assemble on Au(111) in a lying-down configuration irrespective of the concentration and temperature. XPS and electrochemical data indicate a DTT surface coverage of theta approximately 0.16 with two S-head-Au covalent bonds per DTT molecule. The DTT monolayer turns the Au surface hydrophilic enough to allow the formation of fluid dimyristoylphosphatidylcholine (DMPC) bilayer domains by vesicle fusion as revealed by in situ atomic force imaging. Methylene blue (MB) and flavin adenine dinucleotide (FAD) have been used as probes to study molecule transport across the bilayer. PMID- 19543606 TI - Bifurcation of self-motion depending on the reaction order. AB - As a simple example of an autonomous motor, the characteristic features of self motion coupled with the acid-base reaction were numerically and experimentally investigated at the air/aqueous interface. Oscillatory and uniform motion were categorized as a function of the reaction order by numerical computations using a mathematical model that incorporates both the distribution of the surface active layer developed from a material particle as the driving force and the kinetics of the acid-base reaction. The nature of the self-motion was experimentally observed for a boat adhered to a camphor derivative with a mono- or di-carboxylic acid on a phosphate aqueous phase as the base. PMID- 19543607 TI - Determining exact molar absorbance coefficients of single-wall carbon nanotubes. AB - A new spray technique coupled with atomic force microscopy is developed to obtain the absolute number of carbon nanotubes in a unit volume. By using the present technique, the absolute molar absorbance coefficient of single-wall carbon nanotubes (SWNTs) has been determined and found to be ca. 2-5 x 10(7) L mol(-1) cm(-1). This molar absorbance coefficient enables us to deal with carbon nanotubes as individual "molecules" and to adequately compare them with other related compounds such as fullerenes. The absorbance coefficient of SWNTs is found to be only 100 times as large as those of fullerenes. This indicates that the large aspect ratio, random orientation in dispersion solution and anisotropic absorption properties of SWNTs substantially reduce their absorption probability. PMID- 19543608 TI - Soft X-ray induced modifications of PVA-based microbubbles in aqueous environment: a microspectroscopy study. AB - We use scanning-transmission X-ray microspectroscopy (STXM) for in situ characterization of the physicochemical changes in air-filled poly(vinyl alcohol) (PVA) based microbubbles upon soft X-ray irradiation. The microbubbles were illuminated directly in aqueous suspension with 520 eV X-rays and a continuous shrinkage of the particles with an illumination time/radiation dose was observed. Utilizing the intrinsic absorption properties of the species and the high spatial resolution of the STXM, the modifications of the particles' structure were simultaneously recognized. A thorough characterization of the microbubble volume, membrane thickness and absorption coefficient was performed by quantitative fitting of the radial transmittance profiles of the targeted microbubbles. Apart from the observed volume contraction, there was no significant change in the shell thickness. The chemical changes in the membranes were clarified via C K edge near-edge X-ray absorption fine structure (NEXAFS) spectroscopy. It was revealed that the observed structural alterations go along with a continuous degradation of the PVA network associated with formation of carbonyl- and carboxyl-containing species as well as an increased content of unsaturated bonds. PMID- 19543622 TI - [Universal neonatal screening: reflections]. PMID- 19543609 TI - ENDOR and HYSCORE analysis and DFT-assisted identification of the third major stable radical in sucrose single crystals X-irradiated at room temperature. AB - Recently, the chemical structure of two of the three major stable radicals (T2 and T3) produced in sucrose single crystals by X-irradiation at room temperature was identified by comparing Density Functional Theory (DFT) calculations of Electron Magnetic Resonance parameters with experimental results [H. De Cooman, E. Pauwels, H. Vrielinck, E. Sagstuen, F. Callens and M. Waroquier, J. Phys. Chem. B, 2008, 112, 7298-7307]. Ambiguities concerning an unusual proton hyperfine coupling (HFC) tensor prevented the identification of the third major stable radical (T1). In the present work, experimental results of continuous wave Electron Nuclear Double Resonance experiments on sucrose single crystals and Hyperfine Sublevel Correlation Spectroscopy experiments on sucrose powder are presented that lift these remaining ambiguities. Using the final set of experimental HFC tensors and employing advanced DFT calculations, the chemical structure of the T1 radical is established: an allylic-type radical with approximately half of the spin density localised on the C2' carbon of the fructose unit, involving glycosidic bond cleavage at the fructose side and a concerted formation of a carbonyl group at the C1' carbon. The electronic structure of the T1 radical is discussed in more detail by means of additional DFT calculations, yielding a better understanding of the peculiar properties of the unusual proton HFC tensor mentioned above. PMID- 19543623 TI - [Auspicious changes are seen in physicians, professional medical societies, and industry relationships]. PMID- 19543624 TI - [Millennium Development Goals and food insecurity: where is the assessment leading us?]. PMID- 19543625 TI - [It will be a long time to understand and improve medical residences]. PMID- 19543626 TI - [Congenital malformations]. PMID- 19543628 TI - [Analysis of infant mortality from congenital malformations in Argentina during the 2002-2006 period]. AB - OBJECTIVE: To analyze the trend and spatial distribution of infant mortality from congenital malformations in Argentina between 2002 and 2006. MATERIALS AND METHODS: Data were provided by the Ministry of Public Health. Congenital malformations were classified according to the International Classification of Diseases, 10th revision. Early neonatal, late neonatal and postneonatal infant mortality components, congenital malformations subgroups and specific malformations were estimated by departments, provinces, and regions to determine: a) Congenital malformations- related death rate; b) infant mortality rate due to congenital malformations. Both indicators were correlated with the respective departmental latitude/longitude. RESULTS: Infant mortality rate due to congenital malformations was 3.33 per thousand and congenital malformations- related death rate 22.7%. Both indicators showed great spatial variability and did not correlate to latitude/longitude. The lowest infant mortality rate due to congenital malformations and congenital malformations-related death rates were found in Patagonia and the Argentine northeast, respectively. The early neonatal period showed the highest infant mortality rate due to congenital malformations and congenital malformations-related death rates was over 20% in the 3 periods. The Argentine northwest and Argentine northeast showed the lower congenital malformations rates-related death rates in the three components than Centro, Cuyo and Patagonia. CONCLUSIONS: The pattern of infant mortality from congenital malformations in Argentina is similar to that of developed countries, characterized by a decrease of infant mortality rate due to congenital malformations and increase of congenital malformations-related death rates, with a prevailing contribution of heart and nervous system malformations. PMID- 19543629 TI - [Highly active antiretroviral therapy in HIV sero-positive children. Disease progression by baseline clinical, immunological and virological status]. AB - INTRODUCTION: Highly active antiretroviral therapy (HAART) has been administered to children infected with human immunodeficiency virus (HIV) since 1996. This kind of therapy is effective in achieving viral suppression and stopping disease progression but prolonged administration increases the risk of toxic effects, favours the onset of viral resistance and leads to decreased adherence. The aim of the present study was to determine prognostic factors among clinical, immunological and virological parameters at the beginning of HAART. POPULATION AND METHODS: We performed a prospective-retrospective observational analysis of a cohort or 564 HIV+ children assisted in Hospitals of Buenos Aires and Rosario, Argentina, treated with HAART since 1998 (media of treatment: 46.78 months. Range: 2-91 months). Patients were divided in groups according to age (younger or older than one year), and outcome (favourable or unfavourable). Stage, CD4 lymphocytes percentage, CD4 lymphocyte cell count and viral load at the beginning of treatment were analyzed with outcome by means of chi(2) tests, and logistic regression. RESULTS: No differences were observed on the percentage of CD4 T cells and viral load at baseline, between children under one year of age with good (n= 79) or bad outcomes (n= 4). Among older children (450 with good outcome, 31 with unfavourable), the following were identified as predictors of bad outcome: HAART initiation during stage C (p= 0.006), CD4 T-cell percentage below 15 percent (p< 0.001) and CD4 absolute value below 500 cells/mm(3) (p= 0.003). CONCLUSIONS: Children older than one year will have better outcome when HAART is initiated before stage C, with more than 15% CD4 or more than 500 cells/mm(3). PMID- 19543630 TI - [Hunger perception and stunting among children living in poor conditions from the north of Argentina]. AB - INTRODUCTION: Nutritional status depicts not only the nourishing balance but also the conditions of life. Proper food availability, in form and amount, are aspects of food security. OBJECTIVE: To determine the relationships between hunger perception and stunting. METHOD: Stratified and multiple step sample from nine provinces from the north of Argentina (n= 3630, children between 6 months to 6 years old living under poor conditions). Hunger perception was explored according to the methodology of the Service of Economic Investigation of the Department of Agriculture of the United States. Row data were converted to z scores and compared against Argentinean growth standard. The comparison took into account the -2 standard deviation cut of point and the standardized prevalence of low height. RESULTS: 69.5% of the population experienced hunger, varying severe hunger between 38.0 to 48.0% according to provinces. The prevalence of stunting (< - 2SD) was between 10-15%. There was association between perception of hunger, height distribution and the standardized prevalence of low height. There was no association between jurisdiction and perception of hunger. CONCLUSION: For children, food insecurity conditions and structural poverty conditions were associated with stunting. PMID- 19543631 TI - [Neonatal cardiac surgery: assessment and comparison of surgical results using the RACHS-1 risk adjustment method]. AB - INTRODUCTION: The RACHS-1 method (Risk Adjustment for Congenital Heart Surgery) is widely used to predict mortality and risk adjustment in pediatric cardiovascular surgery and constitutes a valid tool to compare results among different health centers. OBJECTIVE: To analyze if the mortality observed in the neonatal group is related to age and/or the risk stratification according to RACHS-1. METHODS: From March 2001 to May 2008 we operated on 751 consecutive patients: 160 neonates (0-30 days), 309 infants (31 days-1 year) and 282 olders (1-18 years). Patients in each group were analyzed according to age, RACHS-1, and mortality. We used a logistic regression in which the mortality was the dependent variable and the age and RACHS-1 the independent variables. RESULTS: The total crude mortality was 4.3%, the neonatal 9.2%. We observed a significant statistical difference of RACHS-1 distribution according to age (chi(2)= 219, p< 0.0001). Logistic analysis showed no statistical difference of mortality (p> 0.05) in the age groups compared to RACHS-1. Furthermore, RACHS-1 is a most powerful mortality predictor (p< 0.001) while age is not (p= 0.8). Using our unit one of RACHS-1 as control group, the odds ratio of the different ages were 2.1 (CI 95%: 1.6-2.7) for each RACHS group. CONCLUSIONS: The age of surgery was not an independent risk factor as to mortality. The RACHS-1 method appeared as a powerful risk factor predictor of mortality; no differences were found in the age groups when classified by RACHS -1. PMID- 19543632 TI - [Near drowning in a pediatric population: epidemiology and prognosis]. AB - INTRODUCTION: Submersion injury is associated with high morbidity and mortality, being the third leading cause of accidental death among children. OBJECTIVES: To analyze and describe risk factors, prognosis, and survival of victims, admitted to a third level Community Teaching Hospital. Population, material and methods. A retrospective, observational, analytical study. We studied patients admitted to the pediatric critical care unit, between 06/2000 and 01/2008. The following variables were analyzed: age, sex, length of stay, days of mechanical ventilation, Glasgow Coma Scale (GCS) score, apnea, bradycardia; baseline, 24 and 48 h lactacidemia, submersion time, swimming pool watchers of the victims. Stata 8.0 software was used; continuous variables were analyzed using Wilcoxon test; for categorical variables Z test and Chi square test were used, and a logistic regression analysis was performed. RESULTS: 30 near-drowning victims were admitted, median age was 25 months (R = 11-144 months). 41.3% occurred during summer, 60% were under parental supervision. Sibling supervision was associated with an increased risk of near-drowning (RR: 2.1; 95% CI 1.1-3.2). Immersion time was > 10 minutes in 3.4%; 26% had apnea, and the GCS score was < 5 in 19.99%. Lactic acid at admission was > 3 mmol/l in 10 patients. Risk factors like glucose level > or = 300 mg% (OR: 3.325), apnea (OR: 2.752), bradycardia (OR: 4.74), GCS <5 (OR: 3.550) and inmersion time > 10 minutes (OR: 5.12), were associated with poor prognosis. Mortality was 2/30 patients. CONCLUSION: In our population, the presence of apnea, bradycardia, GCS <5, glucose level > or = 300 mg%, submersion time > 10 minutes, and lactic acid > 6 mmol/l at admission and the first 24 h, were associated with a poor prognosis and serious injury. PMID- 19543633 TI - [Effects of sleep deprivation on medical performance of pediatric residents]. AB - INTRODUCTION: The performance of medical residents can be adversely affected by sleep deprivation, which happens as a consequence of extended work hours. OBJECTIVE: To assess the effects of sleep deprivation on quick reaction, attention, and memory of pediatrics residents after a duty day. POPULATION, MATERIAL AND METHOD: A before and after, descriptive study was conducted. 44 medical residents, distributed in two groups, were included. The first group (23 residents) was subject to two evaluations, one on a non-duty day and then another on an on-duty day. The second group was evaluated only after a duty day and it was used to assess the learning effect. All participants took: a questionnaire including sleep hours, Epworth sleepiness scale, Maslach burnout inventory, Trail making test, PASAT test, digit span, Signoret memory battery and reaction time test. The degree of deterioration in the tests was assessed after an on-duty day. RESULTS: The average sleep hours on the on-duty day was 3 h. A significant deterioration was evidenced in the visual reaction speed tests. Attention, and memory were not affected after the duty hours. CONCLUSION: The speed of reaction is deteriorated by the sleep deprivation experienced by medical residents: PMID- 19543634 TI - [Etiologic evaluation of genetically-caused mental retardation. Diagnostic algorithm and new molecular techniques]. AB - Mental retardation affects 1-3% of the population. Its etiology is heterogeneous and approximately 47% of cases are caused by genetic factors. The aim of this paper is to report on etiologies of mental retardation, to present updates on new technologies of molecular diagnosis, and to analyze their limitations for the appropriate and rational use of them. Finally, an algorithm based on genetics is suggested for the study of mental retardation by reporting on the techniques available in Argentina and in developed countries. A well-defined etiology will lead to the proper management of children with mental retardation, and suitable family counseling. PMID- 19543635 TI - [Loxoscelism: report of a viscerocutaneous case with favorable resolution]. AB - Loxoscelism is a disease caused by the bite of spiders belonging to Loxosceles genus, represented in our country by L. laeta. It is characterized by necrosis of the skin, accompanied in variable percentage by systemic manifestations that can lead to the death. The objective of this presentation is to report the case of a 6-year-old patient who developed visceral cutaneous loxoscelism with favourable resolution. PMID- 19543636 TI - [Tinea corporis: Description of the case presented in the section "What is your diagnosis?" in the previous issue]. PMID- 19543638 TI - [Consensus on drowning prevention: "The child and the water". 1st part]. PMID- 19543639 TI - [Thiopurine S-methyltransferase gene polymorphism in Chilean blood donors]. AB - BACKGROUND: Thiopurine S-methyltransferase (TPMT) is a cytosolic enzyme that catalyzes the S-methylation of 6-mercaptopurine and azathioprine. Low-activity phenotypes are correlated with polymorphism in the TPMT gene. Patients with low or undetectable TMPT activity could develop severe myelosuppression when they are treated with standard doses of thiopurine drugs. Since ethnic differences in the TPMT gen polymorphism have been demonstrated worldwide, its assessment in the Chilean population is worthwhile. AIM: To investigate the TMPT gene polymorphism in a Chilean blood donor individuals. SUBJECTS AND METHODS: The frequency of four allelic variants of the TPMT gene, *2 (G238C), *3A (G460A and A719G), *3B (G460A) and *3C (A719G) were analyzed in 210 Chilean blood donors, using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and allele specific PCR-based assays. RESULTS: TPMT variants associated to low enzymatic activity, were detected in 16 subjects (8%), who had a heterozygous genotype (*3A in 12; *3C in three and *2 in one subject). No TPMT*3B allelic variant was found. The normal allele (wild-type) was found in 92% of studied individuals. CONCLUSIONS: The allele TPMT*3A, is the most prevalent in this group of Chilean blood donors, as in Caucasian populations. PMID- 19543641 TI - [Health related quality of life among patients on chronic hemodialysis]. AB - BACKGROUND: Health-related quality of life (HRQOL) among patients on chronic hemodialysis (CHD), is associated with mortality, complications and compliance to treatment. AIM: To assess HRQOL in a group of patients on CHD. PATIENTS AND METHODS: A cross-sectional multicenter study was carried out, involving 224 patients from five CHD units (3 private and 2 public) in Bio Bio Region, using the Kidney Disease Quality of Life -36 items (KDQOL-36) questionnaire and Karnofsky scale. Scores range from 0 to 100, with higher values representing a better HRQOL. RESULTS: Physical and Mental scales and subscales of symptoms, effect and the burden of kidney disease subscales rendered scores below 50 (the referential value), in 80%, 61%, 8%, 43% and 80% of evaluations, respectively. The lower scores were observed in patients with diabetes, coronary artery disease, hypoalbuminemia, serum creatinine below 9.4 mg/dL, age >or=55 years and in those with a low economic and educational level (p <0.05). CONCLUSIONS: HRQOL in patients on hemodialysis had values below the referential score in subjects with diabetes and coronary artery disease, poor nutritional status and a low educational and socioeconomic level. The incorporation of support professionals, such as social workers, psychologists, dieticians, covering psychosocial factors, could improve the patients quality of life. PMID- 19543640 TI - [Clinical and metabolic features of subjects with glucose intolerance and high fasting glucose levels]. AB - BACKGROUND: Subjects with glucose intolerance or high fasting glucose levels have a higher cardiovascular risk and frequently become diabetic. AIM: To assess clinical and metabolic characteristics of patients with glucose intolerance or high fasting glucose levels. MATERIAL AND METHODS: Fasting and post glucose load serum glucose and insulin levels were measured in 1404 people, aged 42,0 +/- 14,2 years (81% women) with high diabetic risk. We categorized subjects in different alterations of blood glucose, according to 2006 American Diabetes Association categories. Insulin resistance (RI), insulin secretion (beta %) and insulin disposition (ID), were calculated using fasting blood glucose and insulin levels, using the homeostasis model assessment (HOMA I and II). RESULTS: Sixty percent of studied subjects had first grade relatives with diabetes mellitus and 1097 (78%) were categorized as normal (N), 45 (3%) as Diabetes Mellitus (DM), 161 (11%) as high fasting glucose levels (GAA) and 103 (7%) as glucose intolerant (ITG). Fifty three of the 106 subjects with GAA (50%), were also glucose intolerant. Subjects with GAA had similar insulin sensitivity and lower beta cell function than N (insulin disposition 58 +/- 12 and 111 +/- 32%, respectively p < 0.01). ITG had less insulin sensitivity than N (HOMA-IR 2.6 +/- 1.50 +/- and 2.0 +/- 1.30, respectively) and only a mild decrease in beta cell function (insulin disposition 96 +/- 26 and 111 +/- 32% respectively, p < 0.01). Patients GAA plus ITG had similar alterations than those with DM (HOMA-IR 3.8 +/- 2.2 and 4.4 +/- 3.7 respectively; insulin disposition 57 +/- 10 and 56.0 +/- 26% respectively. CONCLUSIONS: Patients with higher fasting glucose levels behave differently from those with glucose intolerance. High fasting glucose levels are highly prevalent in subjects with high risk of DM and must be considered as risk indicator in preventive programs for diabetes mellitus. PMID- 19543642 TI - [Pathogenicity island region of clinical and environmental strains of Vibrio parahaemolyticus, isolated in Chile]. AB - BACKGROUND: Most clinical isolates of Vibrio parahaemolyticus produce a major virulence factor known as the thermostable direct hemolysin (TDH). TDH is encoded by the tdh gene which is located in a genomic pathogenicity island (PAI). Most environmental isolates are described as tdh negative. AIM: To assess if environmental strains lack the full pathogenicity island or if only the tdh gene is deleted. MATERIAL AND METHODS: Thirty eight clinical and 66 environmental strains of Vibrio parahaemolyticus were studied. PAI was characterized by polymerase chain reaction (PCR). The presence of tdhA and tdhS genes, was determined by Southern blot. RESULTS: Fifty three environmental strains (80%) lacked a full PAI when compared with clinical strains. In environmental strains, Southern blot and sequence analysis showed that a genetic region of 80 kilobase pairs including genes from VPA1310 to VPA1396 was missing. CONCLUSIONS: These results highlight the genetic dynamism of Vibrio parahaemolyticus pathogenecity island region and suggest that new pathogenic strains could appear by horizontal transfer of the island between toxigenic and non-toxigenic strains. PMID- 19543643 TI - [Changes in asthma prevalence among school children during a 6 -year period: Influence of socioeconomic status]. AB - BACKGROUND: Asthma prevalence is increasing worldwide, especially in developing countries. AIM: To determine the prevalence of asthma diagnosis and related symptoms in children, its changes during a 6-year period and the influence exerted by socioeconomic status. MATERIALS AND METHODS: Cross-sectional study conducted among school children of Santiago. A sample of 4,561 children aged 6-7 and 13-14 years attending public schools (central district, medium-low and low socioeconomic status) and private schools (eastern districts, high socioecomic status) were surveyed. Dependent variables were asthma diagnosis (AD), wheezing during the last 12 months (WHE12) and frequency of episodes of wheezing (FEW). Prevalence odds ratios were calculated using regression models, adjusted for several confounding variables. RESULTS: AD prevalence for the group as a whole was higher in boys than in girls (13.2% and 10.8%, respectively p =0.016). Prevalence rate of AD was also higher for children aged 13-14 than 6-7 years (13.8% and 10.1%, respectively p <0.01). An inverse association was found between SES and WHE12. Higher maternal education level was associated with lower prevalence of WHE12 in both age groups and with lower prevalence of FEW in 6-7 year-old children. Paternal education level was positively associated with AD among younger children. At the central district, increasing trends of WHE12 and FEW prevalence were observed during 1994-2000 among 6-7 years children. CONCLUSIONS: A significant association was observed between socioeconomic status, asthma diagnosis and symptoms in school children living in Santiago. Data support an increasing trend of asthma prevalence and severity among 6-7 year-old children during the period 1994-2000. PMID- 19543644 TI - [Familial risk factors for suicide among adolescents with depression]. AB - BACKGROUND: Suicide is the second or third cause of death among Chilean teenagers aged 15 to 19 years. AIM: To evaluate familial risk factors associated to suicidal attempt in adolescents with a depressive disorder. MATERIAL AND METHODS: Thirty two teenagers with depression, aged between 13 and 18 years, receiving psychological treatment in six public Mental Health Centers in Santiago and 32 fathers, mothers or tutors were evaluated. Beck's Depression Inventory and the Family Adaptability and Cohesion Evaluation Scale (FACES III, family version) were applied to adolescents. Adults were evaluated by the Goldberg's General Health Questionnaire and the FACES III parents version. RESULTS: No meaningful statistical association between adolescents suicidal attempt and family risk factors (family order, presence of family stressors and parent's mental health), was observed. However, there was a significant association between suicidal ideation of teenagers, their familiar adaptability and suicidal ideation of close relatives. CONCLUSIONS: Suicidal attempt in this group of teenagers was associated with familial adaptability and suicidal ideation by close relatives. Therefore close relatives must be integrated to the prevention programs for adolescents' suicide. PMID- 19543645 TI - [Surgery for pulmonary tuberculosis. Review of 33 operated patients]. AB - BACKGROUND: Surgical treatment for pulmonary tuberculosis is mainly limited to the management of sequelae such as bronchiectasis, hemoptysis and brochopleural fistulae. AIM: To review the data of patients who underwent surgical treatment for pulmonary tuberculosis. MATERIAL AND METHODS: Retrospective review of 33 patients aged 18 to 73 years (24 males) who underwent lung resection surgery for the management of pulmonary tuberculosis. Follow-up data were obtained from outpatient visit records and registries of the national tuberculosis program. RESULTS: The reasons to perform surgery were the following: fifteen for hemoptysis, nine for lung destruction and nine for an active and multiresistant disease. No patient died in the postoperative period. The morbidity observed included empyema (n =5), pneumothorax (n =2), bronchopleural fistula (n =2) and hemothorax (n =2). At six months of follow up, six of the nine patients with active tuberculosis had negative acid-fast bacilli on sputum smear. Two of these patients died, one due to respiratory failure and another by an unrelated cause. Both dead patients had negative acid-fast bacilli on sputum smear. CONCLUSIONS: Surgery in pulmonary tuberculosis has a high rate of complications but may be useful in selected patients. PMID- 19543646 TI - [Effectiveness of a repellent paint against the spider Loxosceles laeta]. AB - BACKGROUND: Loxoscelism is a severe reaction to the bite of the spider Loxosceles laeta. In recent years, a paint with repellent properties has been promoted in the commerce. However, there are no reports of experiments evaluating its effectiveness. AIM: To evaluate experimentally the repellent properties of a paint against Loxosceles laeta. MATERIAL AND METHODS: Males, females and nymphs of L laeta were deposited in cockpits that allow the free displacement of the spider. Half of the cockpit was covered with repellent paint. Daily observations during one week, determined how frequently the spiders occupied the space covered with repellent paint. The experiments were run in triplicate. RESULTS: No statistical differences in the occupancy of spaces covered with repellent paint or not covered with it were observed for nymphs (87% and 67%, respectively), males (72% and 77%, respectively) or females (91% and 84%, respectively). CONCLUSIONS: The tested paint does not have a repellent action against the spider Loxosceles laeta. PMID- 19543647 TI - [Problem based learning from the perspective of tutors]. AB - BACKGROUND: Problem based learning is a student centered learning technique that develops deductive, constructive and reasoning capacities among the students. Teachers must adapt to this paradigm of constructing rather than transmitting knowledge. AIM: To interpret the importance of tutors in problem based learning during a module of Health research and management given to medical, nursing, physical therapy, midwifery, technology and nutrition students. MATERIAL AND METHODS: Eight teachers that participated in a module using problem based learning accepted to participate in an in depth interview. The qualitative analysis of the textual information recorded, was performed using the ATLAS software. RESULTS: We identified 662 meaning units, grouped in 29 descriptive categories, with eight emerging meta categories. The sequential and cross generated qualitative analysis generated four domains: competence among students, competence of teachers, student-centered learning and evaluation process. CONCLUSIONS: Multiprofessional problem based learning contributes to the development of generic competences among future health professionals, such as multidisciplinary work, critical capacity and social skills. Teachers must shelter the students in the context of their problems and social situation. PMID- 19543648 TI - [Restless legs syndrome is highly underdiagnosed in a neurologic-psychiatric outpatient clinic]. AB - BACKGROUND: Restless legs syndrome (RLS) is a neurological condition that is characterized by the irresistible urge to move the legs and is very common. In the last decade, much attention has been focused on RLS, given its high occurrence, underdiagnosis, and impact on quality of life. AIM: To determine the frequency of RLS in a neurologic-psychiatric outpatient clinic. PATIENTS AND METHODS: We interviewed patients attending a private neurological outpatient clinic, using a standardized validated questionnaire, and an additional phone interview to confirm diagnosis. RESULTS: Of approximately 800 people attending the clinic, the questionnaire was answered by 238 subjects (168 females). Fifteen percent of respondents were affected by RLS and none had been diagnosed before. Most patients had a severe form that probably required treatment. CONCLUSIONS: A low awareness of RLS exists in Chile, even among specialized physicians. PMID- 19543649 TI - [Small bowel transplantation: Report of a single case]. AB - Small bowel transplantation is associated with a patient survival at one and five years, of 80% and 63%, respectively. We report a 36 year-old female with short bowel syndrome, subjected to the first small bowel transplantation performed in Chile. A cadaveric gran was used. Immunosuppression was achieved by means of alemtuzumab, tacrolimus, sirolimus, micofenolate mofetil and steroids. Serial endoscopies and biopsies were performed during seven months after transplantation. The most important late complications were a drug induced renal failure, infections caused by opportunistic agents and a gastrointestinal bleeding probably induced by drugs. After 29 months of follow up, the patient is ambulatory, on oral diet only and with no evidence of graft rejection. PMID- 19543650 TI - Morbid obesity in an adolescent with Prader-Willi syndrome. AB - Prader-Willi syndrome is an uncommon multisystem genetic disorder caused by defects of chromosome 15 (15qll-ql3), often due to deletions or uniparental disomy The syndrome is characterized by neonatal hypotonia, dysmorphic facial features, short stature, motor and mental disabilities, behavioral changes, hyperphagia, precocious obesity and hypogonadotropic hypogonadism. We present a 17 year-old woman, with a previous genetic diagnosis of Prader-Willi syndrome and BMI of 74 Kg/m(2), that was admitted in anasarca, with marked cyanosis, dyspnea and oliguria. She presented high levels of blood urea, creatinine and aminotransferases, in addition to hyperkalemia and hyperuricemia. She had been in regular use of fluoxetine during the last six months, and evolved with severe high blood pressure and respiratory failure, which needed intensive care support. Moreover, sequels and clear signs of recent self-injuries were observed in her trunk, forearms and hands. The findings of morbid obesity, anasarca, self-injury, hyperuricemia and hypoxemia in Prader-Willi syndrome are emphasized. PMID- 19543651 TI - [Histiocytic sarcoma of the small intestine: Report of one case]. AB - Histiocytic sarcoma (HS) is a rare malignant neoplasm of the lymphohematopoietic system, that occurs in lymph nodes, skin and at extranodal sites, particularly the gastrointestinal tract. Although it shows characteristics histological and immunohistochemical features, it may be misdiagnosed. We report a 67 year-old female patient presenting with colicky abdominal pain and vomiting. A CT scan of the abdomen revealed a tumor in the ileum, that was surgically removed. On pathology, the neoplastic cells displayed large abundant eosinophilic cytoplasm, with bizarre-shaped nuclei, that expressed CD 45, CD 68 and lisozyme. The diagnosis of HS requires the use of a panel of immunohistochemical markers and may be supported by ultrastructural findings. PMID- 19543652 TI - [Osteonecrosis of the jaw secondary to oral alendronate: Report of three cases]. AB - Osteonecrosis of the jaw associated to biphosphonate use is more common in cancer patients with bone metastases, that are using intravenous diphosphonates. When these drugs are used orally the risk of the complication is lower. We report 3 diabetic women aged 69, 76 and 82 years, receiving alendronate 70 mg every one week. The unveiling event was the extraction of several teeth without the use of antibiotics. All had bone pain, purulent discharge, loss of bone and halitosis. All improved five months after discontinuing alendronate. PMID- 19543653 TI - [Novel treatments for hepatitis C viral infection and the hepatic fibrosis]. AB - Hepatitis C virus (HCV) infection represents a global health problem due to its evolution to hepatic cirrhosis and hepatocellular carcinoma. The viral pathogenesis and infectious processes are not yet fully understood. The development of natural viral resistance towards the host immune system represents a mayor challenge for the design of alternative therapeutic interventions and development of viral vaccines. The molecular mechanisms of hepatic fibrosis are well described. New alternatives for the treatment of patients with HCV infection and hepatic cirrhosis are under intensive research. New drugs such as viral protease inhibitors and assembly inhibitors, as well as immune modulators have been studied in clinical trials. Additional alternatives include antifibrotic drugs, which reverse the hepatic cellular damage caused by HCV infection. This review makes reference to viral infective mechanisms, molecular pathways of liver fibrosis and overviews conventional and new treatments for HCV infection and liver fibrosis. PMID- 19543655 TI - [Biology of aging]. AB - The study of biological aging has seen a spectacularly fast progress in the last decade. Besides a better understanding and comprehension of physiological aspects, an important advance has been the identification of at least a hundred different genes which control the process of aging. Their mechanism of action falls within the expectations from a handful of theories which attempt to provide a global explanation of the phenomenon of aging, including free radicals, cell senescence and loss of regenerative capacity through the activation of stem cells. In this review we will concentrate in these biological aspects, with a special emphasis on animal models used to study both the genetics and physiology of aging as well as experimental approaches to test the aforementioned theories. It should be emphasized that, while the emphasis is in purely biological aspects of the process, the fast pace of aging of the world's population, including Chile, needs a rapid advance also in our understanding o fits social and economic implications. PMID- 19543654 TI - [Prevention and treatment of Alzheimer disease]. AB - The pharmacological interventions for Alzheimer disease should be based in its pathogenic mechanisms such as amyloidogenesis, tau hyperphosphorilation, disturbances in neurotransmission and changes in neuronal trophism. Other therapies derive from epidemiological observations, such as antioxidants and anti inflammatory drugs, estrogens, statins and anti hypertensive drugs. Some life style interventions, such as changes in diet, exercise and brain stimulation could also be beneficial for the prevention of Alzheimer disease. Ongoing research on pathogenic mechanisms promises the discovery of more effective therapies. Healthy life style should always be recommended due to its benefit and lack of untoward effects. PMID- 19543656 TI - [Systematic reviews of studies of diagnostic test accuracy]. PMID- 19543658 TI - [Latin American citation of Revista Medica de Chile]. PMID- 19543659 TI - Outcome of motor training programmes on arm and hand functioning in patients with cervical spinal cord injury according to different levels of the ICF: a systematic review. AB - OBJECTIVE: To investigate the outcome of motor training programmes on arm and hand functioning in patients with cervical spinal cord injury according to different levels of the International Classification of Functioning, Disability and Health (ICF). DESIGN: Systematic review. METHODS: A search of the following databases: Medline, Cochrane, Cumulative Index to Nursing and Allied Health Literature (CINAHL), EMBASE, Physiotherapy Evidence Database (PEDro) and Database of Abstracts of Reviews of Effects (DARE) from 1976 to August 2008 was performed using the following MeSH terms: Spinal Cord Injuries, Quadriplegia, Rehabilitation, Physical Education and Training, Exercise, Patient-Centered Care, Upper Extremity, Activities of Daily Living, Motor Skills, Motor Activity, Intervention Studies, Clinical Trial. The methodological quality of the selected articles was scored with the Van Tulder Checklist. Descriptive analyses were performed using the PICO principle (Patient characteristics, Intervention, Comparison, Outcome & results) along the ICF function and activity level. RESULTS: Twelve studies were included in the analyses. Overall, the methodological quality of the studies was acceptable, with a mean Van Tulder score of 9.58. Interventions included motor training programmes at the level of function, activity or a combined programme. Most studies reported improvements in arm and hand functioning at the level that was trained for. CONCLUSION: Motor training programmes may improve arm and hand functioning at function and/or activity level in cervical spinal cord injured patients. However, no general conclusion based on a meta-analysis can be drawn due to the wide variety of approaches. PMID- 19543660 TI - Problematic aspects of faecal incontinence according to the experience of adults with spina bifida. AB - OBJECTIVE: To describe problematic aspects of faecal incontinence according to the experience of adults with spina bifida. DESIGN: Qualitative interview study. SUBJECTS: Eleven adults with spina bifida and bowel problems. METHOD: Semi structured open-ended interviews and qualitative analysis. RESULTS: Problematic aspects were related to participation in terms of time consumption, communication, social isolation, love and sexuality, and accessibility. Also, to a sense of helplessness in terms of panic and worry, to a sense of shame in terms of impurity, social acceptance and self-image, and to bowel function in terms of decisions about colostomy, voluntary constipation, and changing patterns. CONCLUSION: The results reveal aspects relevant to supporting clinical practice and suggesting issues for questionnaire studies. PMID- 19543661 TI - Perceived functioning and disability in adults with myotonic dystrophy type 1: a survey according to the International Classification Of Functioning, Disability and Health. AB - OBJECTIVE: The purpose of this study was to describe and analyse self-rated perceived functioning, disability and environmental facilitators/barriers with regard to disease severity, using the International Classification of Functioning, Disability and Health (ICF) checklist, in adults with myotonic dystrophy type 1. DESIGN: Cross-sectional design. SUBJECTS: Forty-one women and 29 men with myotonic dystrophy type 1. METHODS: A modified ICF checklist was used for self-rating of perceived problems in 29 body-function categories, difficulties in 52 activity and participation categories, and facilitators/barriers in 23 environmental-factor categories according to the verbal anchors of the ICF qualifiers. Disease severity classification was based on the muscular impairment rating scale. RESULTS: Of the persons with myotonic dystrophy type 1, 80% perceived problems of excessive daytime sleepiness, 76% of muscle power, and 66% of energy and drive functions, while over 59% perceived difficulties in physically demanding mobility activities. Disabilities in mobility, self-care and domestic life were more frequently reported by persons with severe disease. Support from the immediate family, medicines and social security services were perceived as facilitators for 50-60% of the participants. CONCLUSION: Disabilities and important environmental facilitators in adults with myotonic dystrophy type 1 were identified, and this clinically-relevant information can be used for developing health services for people with this condition. PMID- 19543662 TI - Community integration following moderate to severe traumatic brain injury: a longitudinal investigation. AB - OBJECTIVE: To evaluate the course and identify determinants of community integration for up to 3 years following moderate to severe traumatic brain injury. DESIGN: Prospective cohort study. PATIENTS: A total of 119 moderate to severe traumatic brain injury patients aged 16-67 years. METHODS: The Community Integration Questionnaire was completed at 3, 6, 12, 18, 24 and 36 months post injury. Repeated measures analysis of variance was performed to determine changes over time in the Community Integration Questionnaire and its subscales. Bivariate and multivariate regression analyses were used to identify determinants of community integration 36 months post-injury. RESULTS: Compared with pre-injury, mean home integration, social integration, productivity, and total questionnaire scores decreased 3 months post-injury. Patient scores showed maximal improvement during the first year post--injury. Mean home integration, productivity, and total scores increased to a lesser extent during years 1-3 post-injury. Age, Barthel Index scores, hospital discharge destination, and pre-injury community integration scores were the major determinants of community integration 36 months post-injury (R2 = 60%). CONCLUSION: After an initial decline, mean community integration scores gradually improve following moderate to severe traumatic brain injury. Understanding the course and determinants of community integration is necessary in order to determine functional prognosis following traumatic brain injury. PMID- 19543663 TI - Goal attainment scaling: does it provide added value as a person-centred measure for evaluation of outcome in neurorehabilitation following acquired brain injury? AB - OBJECTIVE: To compare goal attainment scaling (GAS) and standardized measures in evaluation of person-centred outcomes in neurorehabilitation. DESIGN: A prospective cohort analysis from a tertiary inpatient neuro-rehabilitation service for younger adults with complex neurological disability. SUBJECTS/PATIENTS: Consecutive patients (n = 164) admitted for rehabilitation following acquired brain injury (any cause) over 3 years. Mean age 44.8 (standard deviation 14.4) years. DIAGNOSIS: 66% strokes, 18% trauma, 16% other. Male:female ratio 102:62. METHODS: GAS-rated achievement of 1-6 patient-selected goals was compared with the Functional Assessment Measure (UK FIM+FAM), and Barthel Index (BI), rated on admission and discharge. Personal goals were mapped retrospectively to the FIM+FAM and International Classification of Functioning, Disability and Health (ICF). RESULTS: Median (interquartile range; IQR) GAS T scores were 50.0 (44.2-51.8) and moderately correlated with changes in FIM+FAM and BI (both rho 0.38 (p < 0.001)). Standardized response means were 2.2, 1.6 and 1.4 for GAS, FIM+FAM and BI, respectively. Of 667 personal goals set, 495 (74%) were fully achieved. Although 413 (62%) goals were reflected by changes in FIM+FAM, over one-third of goals were set in other areas. CONCLUSION: GAS appeared to be more responsive, and captured gains beyond the FIM+FAM, thus providing added value as an adjunct to outcome measurement in patients with complex disability. PMID- 19543665 TI - Pain responses in repeated end-range spinal movements and psychological factors in sick-listed patients with low back pain: is there an association? AB - OBJECTIVE: Repeated end-range spinal movements producing specific pain responses (i.e. centralization or non-centralization) may be used for diagnostic and prognostic purposes. However, possible associations between psychological factors and pain responses have been reported. The aim of this study was to investigate the associations between pain responses in repeated end-range spinal movement tests and psychological factors. DESIGN: Cross-sectional clinical study. PATIENTS: Data from 331 patients sick-listed for 4-12 weeks due to low back pain with or without sciatica. METHODS: Initially the patients completed a questionnaire including questions about psychological factors. Then they underwent a standardized physical test procedure and were classified according to centralized or non-centralized pain response. RESULTS: Statistically significant associations were found between non-centralization and mental distress (p < 0.009) as well as depressive symptoms (p < 0.049). These associations remained present after adjustment for potential confounders by logistic regression: mental distress odds ratio (OR) 1.16 (95% confidence interval (CI) 1.03-1.30) (p = 0.013), depressive symptoms OR 1.23 (95% CI 1.01-1.51) (p = 0.044). CONCLUSION: The pain responses in repeated end-range spinal movements were not independent of psychological factors. Mental distress and depressive symptoms occurred more often among non-centralizers than among centralizers. It is recommended that the possible influence of psychological factors on the result of mechanical testing be accounted for in future studies. PMID- 19543664 TI - Botulinum toxin A for treatment of upper limb spasticity following stroke: a multi-centre randomized placebo-controlled study of the effects on quality of life and other person-centred outcomes. AB - OBJECTIVE: Botulinum toxin is known to relieve upper limb spasticity, which is a disabling complication of stroke. We examined its effect on quality of life and other person-centred perspectives. DESIGN: A multi-centre, randomized, double blind, placebo-controlled study. PATIENTS: Ninety-six patients were randomized (mean age 59.5 years) at least 6 months post-stroke. Mean time since stroke was 5.9 years. METHODS: Patients received either botulinum toxin type A or placebo into the affected distal upper limb muscles on 2 occasions, 12 weeks apart. Assessment was undertaken at baseline, 8, 12, 20 and 24 weeks. The primary outcome measure was the Assessment of Quality of Life scale (AQoL). Secondary outcome assessments included Goal Attainment Scaling (GAS), pain, mood, global benefit, Modified Ashworth Scale (MAS), disability and carer burden. RESULTS: The groups did not differ significantly with respect to quality of life, pain, mood, disability or carer burden. However, patients treated with botulinum toxin type A had significantly greater reduction in spasticity (MAS) (p < 0.001), which translated into higher GAS scores (p < 0.01) and greater global benefit (p < 0.01). CONCLUSION: Although no change in quality of life was demonstrated using the AQoL, botulinum toxin type A was found to be safe and efficacious in reducing upper limb spasticity and improving the ability to achieve personal goals. PMID- 19543666 TI - Is smoking and alcohol consumption associated with long-term sick leave due to unspecific back or neck pain among employees in the public sector? Results of a three-year follow-up cohort study. AB - OBJECTIVE: To investigate the associations between smoking and alcohol consumption, and long-term sick leave due to unspecific back or neck pain among employees in the public sector. DESIGN: A 3-year prospective cohort study. SUBJECTS: Approximately 9000 persons in the public sector in Sweden were invited to participate. Of these, 7533 answered a questionnaire and 6532 were included in the study, classified as having "good health for working". METHODS: New periods of sick leave >or= 28 days were consecutively reported from the employers or the occupational health service during a period of 3 years. Rate ratios were estimated by means of Cox proportional hazard regression model. RESULTS: Smoking was associated with an increased risk of long-term sick leave due to unspecific back or neck pain. Compared with people who have never smoked, "ever smokers" had a higher risk (rate ratio = 1.8, 95% confidence interval: 1.3-2.4). Alcohol consumption tended to be associated with a decreased risk, but the results were not statistically significant. CONCLUSION: Our results suggest that smoking is a risk factor for long-term sick leave due to unspecific back or neck pain. Moderate alcohol consumption tends to have a protective effect, at least among women in the public sector. PMID- 19543667 TI - Pain relief in women with fibromyalgia: a cross-over study of superficial warmth stimulation and transcutaneous electrical nerve stimulation. AB - OBJECTIVE: To compare the effects of portable superficial warmth with transcutaneous electrical nerve stimulation on pain in patients with fibromyalgia. METHODS: The study had a randomized cross-over design. A total of 32 patients with fibromyalgia were randomly assigned to 2 groups. After instruction, the patients treated themselves using a portable device providing superficial warmth (42 degrees C) or a transcutaneous electrical nerve stimulation apparatus. After 3 weeks the patients switched therapy. The patients rated pain intensity on a 0-100 numerical rating scale before and after each treatment. After 6 weeks, patients were questioned concerning therapy preference. RESULTS: There was no difference in level of pain relief when comparing the 2 treatment modes. Median pain intensity in patients using warmth therapy decreased from 77.5 on the numerical rating scale before treatment to 62.5 after treatment and in patients using transcutaneous electrical nerve stimulation from 80 to 62.5. Ten patients reported a reduction of 20 units or more on the numerical rating scale after warmth therapy, as did 10 after transcutaneous electrical nerve stimulation. Seventeen of 32 patients preferred warmth therapy and 10 preferred transcutaneous electrical nerve stimulation. CONCLUSION: Sensory stimulation with superficial warmth or transcutaneous electrical nerve stimulation yielded comparable temporary pain reduction in patients with fibromyalgia. Both procedures are self-administered, safe and in-expensive. PMID- 19543668 TI - Isokinetic characteristics of shoulder rotators in patients with adhesive capsulitis. AB - OBJECTIVE: To demonstrate the muscle strength characteristics of shoulder internal and external rotators and the effects of isokinetic exercise on muscle activity in patients with adhesive capsulitis. DESIGN: Cohort study with control subjects. PARTICIPANTS: Eight patients with adhesive capsulitis and 8 controls. METHODS: Maximal isometric and isokinetic strength tests of shoulder internal and external rotators in the scapular plane were carried out. Muscle activities of the rotators were recorded in resting and during maximal isometrics. Muscle strength variables (peak torque, total work and power) and myoelectric variables (resting root-mean-square amplitude pre- and post-tests and the external-internal rotator co-activity in resting and during isometric contractions) were recorded. A mixed repeated-measure analysis of variance test was used to examine the within group and between-group differences. RESULTS: For affected shoulders, smaller isometric average torque of internal rotators and high-speed peak torque, total work and power of external rotator were observed. The external/internal rotator ratio of peak torque in high-speed testing also exhibited significant decrease. The myoelectric variables showed no significant changes. CONCLUSION: High-speed external rotator strength and isometric internal rotator strength of the affected shoulders were decreased significantly. Isokinetic exercise may not increase the resting muscle activities and co-activity. These results provide a reference in planning muscle strengthening programmes and goals for these patients. PMID- 19543669 TI - Clinical effectiveness of an interdisciplinary pain management programme compared with standard inpatient rehabilitation in chronic pain: a naturalistic, prospective controlled cohort study. AB - OBJECTIVE: To compare the effects of an interdisciplinary pain management programme with those of standard in-patient rehabilitation by comprehensive biopsychosocial self--assessment. METHODS: In this naturalistic prospective controlled cohort study, 164 chronic pain patients who participated in the interdisciplinary pain programme and 143 who underwent standard rehabilitation were assessed using standardized instruments. Effect differences were compared bivariately and analysed by multivariate logistic regression to control for baseline differences in the outcome variables and confounders. RESULTS: On entry into the clinic, the interdisciplinary pain programme patients were younger and showed significantly worse mental and psychosocial health than the standard rehabilitation patients. At discharge, the interdisciplinary pain programme patients reported greater improvement on pain (multivariate p = 0.034), social functioning (bivariate p = 0.009), and in trend in catastrophizing and ability to decrease pain. At the 6-month follow-up, the effects experienced by the standard rehabilitation group were higher on physical functioning, social functioning, anxiety, and life control (multivariate p = 0.013-0.050). CONCLUSION: Intensive interdisciplinary rehabilitation with more behavioural therapies was accompanied by a greater improvement in patients who were severely affected by pain, compared with standard rehabilitation by the end of the stay, but not in the mid-term. Highly resource-consuming patients may benefit from subsequent, individually tailored outpatient care. PMID- 19543670 TI - Development of the rehabilitation patient experiences questionnaire: data quality, reliability and validity in patients with rheumatic diseases. AB - OBJECTIVE: To develop the Rehabilitation Patient Experiences Questionnaire for patients undergoing rehabilitation for rheumatological disorders. METHODS: Development of the instrument was based on literature review and adaptation of the Patient Experiences Questionnaire. The instrument was piloted and then administered in a multicentre cohort study of 12 rehabilitation units. RESULTS: The survey included 435 patients, of which 412 (94.7%) responded to the Rehabilitation Patient Experiences Questionnaire. Following principal component analysis, the initial 27 items were reduced to 18 items and 4 scales: rehabilitation care and organization, information and communication, availability of staff, and social environment. Item--total correlations ranged from 0.77 to 0.87. Cronbach's alpha exceeded the criterion of 0.7, and was 0.87, 0.86, 0.78, and 0.77 for the 4 scales, respectively. Construct validity was supported by correlations between the 4 scales and responses to individual questions, which were largely in the direction as hypothesized. Overall, patients reported good experiences. There were statistical differences across the rehabilitation settings in staff availability (p = 0.001) and social environment (p = 0.002), but no difference in care and organization and information/communication (p > 0.05). CONCLUSION: The 18-item Rehabilitation Patient Experiences Questionnaire is a promising outcome measure of experiences related to rehabilitation in patients with rheumatic diseases across different clinical settings. : PMID- 19543671 TI - Effectiveness of phantom exercises for phantom limb pain: a pilot study. AB - OBJECTIVE: To investigate the effects of phantom limb exercises on phantom limb pain. METHODS: A total of 20 traumatic amputees participated in the study. Ten received phantom exercises and prosthetic training, and 10 were treated with routine prosthetic training and a general exercise programme. Intensity of pain was evaluated using a 10-cm visual analogue scale before therapy and after 4 weeks of therapy. RESULTS: Baseline scores on the visual analogue scale were similar between the groups. Pain intensity decreased in all subjects after 4 weeks of treatment in both groups. According to the visual analogue scale scores at the end of 4 weeks, the phantom exercises group differed significantly from the general exercise group (p < 0.05). CONCLUSION: Phantom exercises appear to be effective in reducing phantom pain, but further research is required to confirm this.The results of this study indicate that phantom exercises can be used safely to alleviate phantom limb pain in lower and upper limb amputees. PMID- 19543673 TI - Reliability of lower limb kinematics, mechanics and energetics during gait in patients after stroke. AB - OBJECTIVE: To assess the reliability of kinematic, mechanical and energetic gait variables at short (1 day) and medium (1 month) intervals in adult patients after stroke. DESIGN: Prospective study. SUBJECTS: Ten patients with chronic post stroke (mean age 53.5 years; age range 25-80 years). METHODS: Three-dimensional gait analysis was performed 3 times in these subjects: at baseline (T0), after 1 day (T1) and after 1 month (T2). The reliability of the gait analysis was tested by comparing gait variables measured at T1 and T0 (1 day interval), at T2 and T0 (1 month interval). The inter-session reliability of kinematic, mechanical and energetic variables was calculated by intra-class correlation coefficient (ICC). RESULTS: The reliability of kinematic variables ranged from excellent to moderate (ICC >or= 0.51), except for the ankle position at heel strike (ICC = 0.44). The reliability of mechanical and energetic variables ranged from excellent to good (ICC >or= 0.71). The most reliable variable was external mechanical work (ICC = 0.96). The kinematic, mechanical and energetic variables did not change significantly between T0, T1 and T2 (repeated-measures analysis of variance). CONCLUSION: Kinematic, mechanical and energetic gait variables present good reliability when measured at 1 day and 1 month intervals in adult patients after stroke. PMID- 19543672 TI - Safety of methylphenidate following traumatic brain injury: impact on vital signs and side-effects during inpatient rehabilitation. AB - OBJECTIVE: The aim of the present study was to evaluate the safety of methylphenidate administered during inpatient rehabilitation following traumatic brain injury. METHODS: Forty inpatients with moderate-severe traumatic brain injury (mean 68.4 days post-injury) participated in a randomized, cross-over, double-blind, placebo-controlled trial of methylphenidate administered at a dose of 0.3 mg/kg body weight twice daily. RESULTS: Methylphenidate administration resulted in a statistically significant increase in pulse of 12.3 beats/min (95% confidence interval (CI) 9.25-15.36), diastolic blood pressure of 4.1 mmHg (95% CI 2.11-6.10), and mean arterial pressure of 3.75 mmHg (95% CI 1.79-5.72). These changes did not, however, appear to be symptomatic, as no participants were withdrawn due to adverse events, and there was no significant self-report of increased heart rate with methylphenidate. Blinding was successful. Significantly greater reporting of irritability of 0.14 points (95% CI 0.02-0.26), difficulty sleeping of 0.17 points (95% CI 0.02-0.31) and total side-effects of 0.68 points (95% CI 0.06-1.30) was associated with methylphenidate compared with placebo. CONCLUSION: Methylphenidate given at 0.3 mg/kg body weight appears to be safe in the inpatient rehabilitation phase. This trial is registered with the Australian New Zealand Clinical Trials Registry (12607000503426). PMID- 19543674 TI - Inappropriate citations used to describe the pharmacology of botulinum toxin type A preparations in clinical use. PMID- 19543675 TI - Alternative method of suprapubic assistance in operative cystoscopy. AB - Complications of tension-free vaginal tape (TVT) procedures have become more prevalent as the procedure has become more popular for the treatment of stress urinary incontinence. A common complication of TVT slings is bladder perforation, which is estimated to occur in 0.7-24% of treated patients. While easily treated if identified intraoperatively, unrecognized bladder perforation generally requires further surgery to correct. We propose a novel minimally invasive technique using the suture passer of the Carter-Thomason CloseSure system for suprapubic assistance during cystoscopic removal of TVT mesh from the bladder. This novel approach allows for the avoidance of an open incision or a larger accessory port placed through the bladder to assist with mesh excision. PMID- 19543676 TI - Conservative versus surgical management of prolapse: what dictates patient choice? AB - INTRODUCTION AND HYPOTHESIS: The aim of this study is to establish whether the presence of severe symptoms influences women's choice of pessaries or surgery for uterovaginal prolapse. METHODS: This is a prospective study using the validated Sheffield Prolapse Symptoms Questionnaire. RESULTS: Women choosing surgery (n = 251) were younger (58 versus 66 years), more bothered by dragging lower abdominal pain (33% versus 25%, P = 0.04), need for vaginal digitation (8% versus 3%, P = 0.02), and incomplete bowel emptying (27% versus 19%, P = 0.01) than women choosing pessaries (n = 429). More women opting for surgery were sexually active (51% versus 29%, P < 0.0001), perceived avoidance of sex due to prolapse (28% versus 17%, P = 0.000), and perceived prolapse interfering with sexual satisfaction as a severe problem (26% versus 15%, P = 0.000). CONCLUSIONS: Nearly two thirds of women with symptomatic prolapse initially opted for conservative management. Women choosing surgery over pessaries for treatment of prolapse describe more severe symptoms related to bowel emptying, sexual function, and quality of life and are bothered by them. PMID- 19543677 TI - Urolithiasis in pregnancy. AB - Urolithiasis is the most common cause of urological-related abdominal pain in pregnant women after urinary tract infection. The disease is not uncommon during pregnancy occurring in 1/200 to 1/2,000 women, which is not different from the incidence reported in the nonpregnant female population of reproductive age. During pregnancy, the frequency of stone localization is twice as higher in the ureter than in the renal pelvis or calyx, but there is no difference between the left and right kidney or ureter. Urinary stones during pregnancy are composed mainly of calcium phosphate (hydroxyapatite) in 74% of cases and calcium oxalate in the remaining 26% (Ross et al., Urol Res 36:99-102, 2008). In conclusion, urolithiasis during pregnancy can be serious, causing preterm labor in up to 40% of affected women. The pathogenesis, diagnosis, and management are analyzed. PMID- 19543678 TI - Polycystic kidney disease channel and synaptotagmin homologues play roles in schizosaccharomyces pombe cell wall synthesis/repair and membrane protein trafficking. AB - Eukaryotic cells can sense a wide variety of environmental stresses, including changes in temperature, pH, osmolarity and nutrient availability. They respond to these changes through a variety of signal-transduction mechanisms, including activation of Ca(2+)-dependent signaling pathways. This research has discovered important implications in the function(s) of polycystic kidney disease (PKD) channels and the mechanisms through which they act in the control of cell growth and cell polarity in Schizosaccharomyces pombe by ion channel-mediated Ca(2+) signaling. Pkd2 was expressed maximally during the exponential growth phase. At the cell surface pkd2 was localized at the cell tip during the G(2) phase of the cell cycle, although following cell wall damage, the cell surface-expressed protein relocalized to the whole plasma membrane. Pkd2 depletion affected Golgi trafficking, resulting in a buildup of vesicles at the cell poles, and strongly affected plasma membrane protein delivery. Surface-localized pkd2 was present in the plasma membrane for a very short time and was rapidly internalized. Internalization was dependent on Ca(2+), enhanced by amphipaths and inhibited by gadolinium. The pkd2 protein was in a complex with a yeast synaptotagmin homologue and myosin V. Depletion of pkd2 severely affected the localization of glucan synthase. A role for pkd2 in a cell polarity and cell wall synthesis signaling complex with a synaptotagmin homologue, myosin V and glucan synthase is proposed. PMID- 19543679 TI - A new approach for adipose tissue treatment and body contouring using radiofrequency-assisted liposuction. AB - A new liposuction technology for adipocyte lipolysis and uniform three dimensional tissue heating and contraction is presented. The technology is based on bipolar radiofrequency energy applied to the subcutaneous adipose tissue and subdermal skin surface. Preliminary clinical results, thermal monitoring, and histologic biopsies of the treated tissue demonstrate rapid preaspiration liquefaction of adipose tissue, coagulation of subcutaneous blood vessels, and uniform sustained heating of tissue. PMID- 19543680 TI - [Cognitive training in rehabilitation: a program to treat mild cognitive impairment]. AB - As part of a research project, a behavior therapy-oriented cognitive training program, designed to improve the cognitive ability of middle-aged employees (50 to 59 years), was implemented and evaluated. The goal of the training program was to analyze the cognitive, affective and behavior-related conditions of memory deficits in the workplace. The training focused on two topics: (1) development of new job tasks, (2) lack of time and deadline constraints. A total of 316 patients in the psychosomatic clinic of Bad Neustadt participated in the study. Those who showed cognitive impairments took part in the cognitive training program during the intervention phase. During the control phase, patients with cognitive impairments received no additional intervention. The neuropsychological screening was repeated with patients with cognitive impairments at discharge. The findings show that the objective and subjective memory performance of the participants improved significantly compared to that of the control group. At the 1-year follow-up, 84% of the participants reported that they could use the training strategies in their job. PMID- 19543681 TI - Predicting in-patient falls in a geriatric clinic: a clinical study combining assessment data and simple sensory gait measurements. AB - BACKGROUND: Falls are among the predominant causes for morbidity and mortality in elderly persons and occur most often in geriatric clinics. Despite several studies that have identified parameters associated with elderly patients' fall risk, prediction models -- e.g., based on geriatric assessment data -- are currently not used on a regular basis. Furthermore, technical aids to objectively assess mobility-associated parameters are currently not used. OBJECTIVES: To assess group differences in clinical as well as common geriatric assessment data and sensory gait measurements between fallers and non-fallers in a geriatric sample, and to derive and compare two prediction models based on assessment data alone (model #1) and added sensory measurement data (model #2). METHODS: For a sample of n=110 geriatric in-patients (81 women, 29 men) the following fall risk associated assessments were performed: Timed 'Up & Go' (TUG) test, STRATIFY score and Barthel index. During the TUG test the subjects wore a triaxial accelerometer, and sensory gait parameters were extracted from the data recorded. Group differences between fallers (n=26) and non-fallers (n=84) were compared using Student's t-test. Two classification tree prediction models were computed and compared. RESULTS: Significant differences between the two groups were found for the following parameters: time to complete the TUG test, transfer item (Barthel), recent falls (STRATIFY), pelvic sway while walking and step length. Prediction model #1 (using common assessment data only) showed a sensitivity of 38.5% and a specificity of 97.6%, prediction model #2 (assessment data plus sensory gait parameters) performed with 57.7% and 100%, respectively. DISCUSSION AND CONCLUSION: Significant differences between fallers and non-fallers among geriatric in-patients can be detected for several assessment subscores as well as parameters recorded by simple accelerometric measurements during a common mobility test. Existing geriatric assessment data may be used for falls prediction on a regular basis. Adding sensory data improves the specificity of our test markedly. PMID- 19543682 TI - [Patient view in the assessment of quality of life in old age: potentials and limits]. AB - Quality of life is a multidimensional construct commonly used in geriatric health care. The patient's view is increasingly incorporated in its determination. Pain and satisfaction with life are two important subjective facets of quality of life. Potentials and limits of an integration of the subjective view in seemingly heavily disadvantaged patient populations are discussed using three approaches: 1) Self-reports on pain in aphasic patients can often be obtained by means of nonverbal communication or verbal communication adapted to the disorder. 2) Even in people suffering from dementia, pain can be assessed in self-reports into the middle stages of the disease. In severe dementia, observational methods have been developed, but their significance with respect to the experience of pain is still being debated in the scientific community. 3) Differences in content and structure in the individual construction of life-satisfaction in multimorbid elderly without cognitive impairment can be reproduced by an individualized measurement tool (FLQM). It allows for determination as well of global and domain specific life-satisfaction as of differential determinants of longitudinal changes. All three approaches highlighted potentials for an extended integration of the patient's perspective in the assessment and evaluation of their quality of life. PMID- 19543683 TI - Achalasia--which method of treatment to choose for senior patients? AB - BACKGROUND: Achalasia is an uncommon illness affecting 1 per 100,000 patients a year. It encompasses a rare, primary motor disorder of the distal esophagus. METHODS: Over the period 1998-2006, 115 patients underwent various treatments for achalasia; the subgroup of seniors consisted of 26 patients. Six patients of these (age 69.7 y) underwent a modified Heller cardiomyotomy due to failure of previous endoscopic interventions. Standard esophageal manometry and 24 hour pH metry were performed pre- and postoperatively. RESULTS: Six senior patients with achalasia underwent a laparoscopic Heller myotomy. Average preoperative tonus of the LES was 55 mmHg, postoperative tonus of the LES decreased to 11 mmHg. We performed Toupet partial fundoplication in all patients; no microperforation of the esophagus was found in the preoperative esophagoscopy. We recorded minimal pathological gastroesophageal reflux in pH metry - the average preoperative DeMeester score was 8, postoperatively 10.5. Prolonged dysphagia was not present in any patient--preoperative GIQLI score was 94, postoperative score was 106. There was no mortality or morbidity in the group of the operated patients. CONCLUSION: Our operational results and postoperative follow-up show that laparoscopic Heller myotomy with Toupet partial fundoplication is a safe and effective treatment and can be recommended as the method of first choice for senior patients with no contraindication for laparoscopic operation. PMID- 19543684 TI - [Relationships between activities of daily living and glucose metabolism in geriatric patients with type 2 diabetes mellitus]. AB - In higher age, various diseases are more frequently present and occur in combination, with diabetes mellitus representing the comorbidity seen most frequently. Within the framework of a retrospective study, it has been evaluated whether the degree of metabolic compensation has an influence on the activities of daily life in geriatric patients after an acute event and following rehabilitation treatment. For this purpose over a period of 27 months (01/02/2000 - 03/31/2002), diabetic patients of a Saxonian geriatric center (n = 644, mean age 77.86 years/SD 7.115, 30.3% males) were examined before, directly after, and 2 - 3 years after rehabilitation. Independent of age, rehabilitation resulted in a significant increase in activities of daily living (ADL, Barthel index); however, after 2 - 3 years these values decreased again, not depending on age, but they remained significantly above the baseline values (multiple comparison of mean values). The values at the time of dismissal did not depend on metabolic parameters. The result of the rehabilitation treatment checked by means of a questionnaire 2 - 3 years after dismissal showed clear advantages for those patients having a near-normal compensation of metabolism. When HbA1c was below 7% at the time-point of the geriatric rehabilitation the Barthel index score in the questionnaire was found to be significantly higher (multiple comparisons of mean values) than in patients with a HbA1c of 7% or higher. With increasing age, the danger of hypoglycemia was seen to be significantly elevated (Fisher's exact test). PMID- 19543686 TI - Current topics in pancreato-biliary endotherapy: what can we do? AB - INTRODUCTION: Endotherapy is progressing steadily, especially for various pancreato-biliary diseases. This article introduced new procedures and devices, and revealed improvement of treatment outcomes. MATERIALS AND METHODS: Biliary covered metallic stent (CMS) has developed, and the indication of CMS placement is changing because of its removability. CMS is effective not only for unresectable biliary malignancies but also for resectable tumors, benign biliary strictures, and benign pancreatic strictures. Drug-eluting CMS can be used as anti-tumor agents. Interventional endoscopic ultrasonography (EUS) has shifted the treatment paradigm because it is possible to approach lesions through the digestive tract wall. The diagnosis and treatment of pancreatic cancer using interventional EUS technique are effective, feasible, and promising. Recently, trans-gastric necrosectomy for an infected pseudocyst was reported as an alternative treatment to surgery. Double- and single-balloon enteroscopy will be performed more frequently to treat the pancreato-biliary disorders in the patients with altered anatomy. Endoscopic papillary large balloon dilation (EPLBD), new procedure to the papilla, can treat large bile duct stones effectively without lithotripsy. CONCLUSION: This paper introduces current developments in endotherapy to surgeons. These procedures are of very great interest because they alter the treatment algorithms for many pancreato-biliary diseases. PMID- 19543685 TI - [Mobility and safety for elderly (MoSi), a new intervention to improve mobility and gait in elderly people]. AB - BACKGROUND: "Mobility and safety for the elderly (MoSi)") is a new intervention, especially designed for elderly people with gait disorders and for fallers. METHOD: 165 persons, 65 years or older, participated in this study.The participants were assessed prior to and after the 5-week intervention. The intervention included various elements of strength and balance training, stretching, reaction and coordination training, and information. The participants were instructed about how to prevent falls and what measures to take after a fall has occurred. Furthermore, they were taught home exercises. The intervention was set in an ambulatory physiotherapy department and consisted of 10 training sessions. RESULTS: Participants reported improved performance of gait (66 %), gait security (66 %), power (65 %), balance (62 %), sense of security (61 %) capacity (70 %) and well being (63 %).The Berg Balance Scale and the Balance test of the "Tinetti Balance Scale" showed significant improvements (p<.001). The Timed Up and Go Test, the gait test of the Tinetti Balance Scale showed improvements in persons with very impaired gait (p<.005). The Repeated Chair Stands Test showed no significant improvements. 137 (95 %) of the participants stated they would continue the exercises at home, 112 (79 %) stated they would continue the exercises under therapeutic supervision. DISCUSSION: The results show that the intervention is able to improve mobility and gait of elderly persons, subjectively and objectively, especially of persons with impaired gait. The motivation effect of the intervention was high which was marked by the large number of participants which wanted to continue their exercises at home. Furthermore, the intervention resulted in participants reporting improvements in other parameters such as capacity and well being. CONCLUSION: The intervention is able to improve important measures of balance in community dwelling older people. However, the effect on the number of falls still has to be investigated. The intervention is suitable for an ambulatory setting and shows significant effects after a short time. Due to the intervention's compactness it is appropriate for persons who normally would avoid training programs because of the duration. PMID- 19543687 TI - The association of pagophagia with Helicobacter pylori infection in patients with iron-deficiency anemia. AB - This study aimed to determine the relationship between pagophagia (compulsive ice eating) and H. pylori infection in patients with iron-deficiency anemia. We identified H. pylori infection using the (13)C-urea breath test in 45 patients with iron-deficiency anemia (group 1) and 55 patients with iron-deficiency anemia and pagophagia (group 2). Subgroups for testing oral intestinal iron absorption were randomly assigned from both groups. These subgroups consisted of (a) 10 patients with iron-deficiency anemia, (b) 10 patients with iron-deficiency anemia and pagophagia, (c) 10 patients with iron-deficiency anemia, pagophagia, and H. pylori infection before the eradication of H. pylori and (d) subgroup c after eradication therapy. There was no difference in the rate of H. pylori infection in the iron-deficiency anemia groups, with or without pagophagia. Furthermore, oral intestinal iron absorption was not influenced by pagophagia and/or H. pylori infection. Pagophagia did not increase the risk of H. pylori infection in patients with iron-deficiency anemia. Pagophagia and H. pylori infection do not synergistically affect the development of intestinal iron absorption abnormalities. PMID- 19543688 TI - Generation and evaluation of an H9N1 influenza vaccine derived by reverse genetics that allows utilization of a DIVA strategy for control of H9N2 avian influenza. AB - H9N2 avian influenza viruses have circulated widely in domestic poultry around the world, and their outbreaks have resulted in heavy morbidity and mortality. In addition, H9N2 avian influenza viruses were transmitted directly from birds to humans in Hong Kong and mainland China during 1998 and 2003, which prompted the public health authorities to seek protective strategies to control H9N2 influenza viruses. In this study, we attempted to develop a DIVA (differentiating infected and vaccinated animals) strategy for H9N2 avian influenza viruses. This strategy does not interfere with serological monitoring and allows effective control of H9N2 avian influenza. We generated a reassortant H9N1 influenza vaccine strain by reverse genetics and employed an enzyme-linked immunosorbent assay (ELISA) with a truncated N1 antigen expressed in E. coli to differentiate between vaccinated and naturally infected animals. Immunization of BALB/c mice with the inactivated reassortant H9N1 vaccine conferred protection against lethal challenge with H9N2 viruses. Meanwhile, the ELISA can be used to distinguish between vaccination and natural infection quickly and easily. Therefore, this study has opened up a new avenue for the control of H9N2 avian influenza. PMID- 19543689 TI - Coadministration of chicken GM-CSF with a DNA vaccine expressing infectious bronchitis virus (IBV) S1 glycoprotein enhances the specific immune response and protects against IBV infection. AB - Various approaches have been developed to improve the efficacy of DNA vaccination, such as the use of plasmids expressing cytokines as molecular adjuvants. The purpose of the present study was to determine whether co administration of a plasmid containing a chicken granulocyte-macrophage colony stimulating factor (GM-CSF) gene and a plasmid containing the S1 gene of infectious bronchitis virus (IBV) could enhance the immune response and protection efficacy in chickens against challenge by virulent IBV. Plasmids carrying the S1 gene of IBV (pVAX-S1) and the chicken GM-CSF gene (pVAX-chGM-CSF) were constructed. Seven-day-old chickens were injected intramuscularly with pVAX S1, pVAX-chGM-CSF, or both and boosted 2 weeks later. Chickens were challenged with virulent IBV at 3 weeks after the booster immunization and observed for 2 weeks. The results showed that co-administration of pVAX-chGM-CSF led to a significant enhancement of humoral and cellular responses over that of vaccination with pVAX-S1 alone. In addition, vaccination with pVAX-chGM-CSF and pVAX-S1 provided 86.7% protection (13/15) against IBV challenge. In contrast, only 73.3% of the chickens were protected against IBV challenge by pVAX-S1 vaccination alone. These results strongly indicate that chGM-CSF can be used as a molecular adjuvant to enhance the protective immunity induced by an IBV-specific DNA vaccine. PMID- 19543690 TI - SUNCT and optic nerve hypoplasia. AB - SUNCT has been reported in association with abnormalities of the brainstem and pituitary region. We present a patient with a history of left optic nerve hypoplasia, mild hypothalamic-pituitary dysfunction, and SUNCT starting in adolescence. SUNCT with an early age of onset may be associated with congenital abnormality of the hypothalamic-pituitary axis. PMID- 19543692 TI - Clinical ethics and the dynamics of group decision-making: applying the psychological data to decisions made by ethics committees. PMID- 19543693 TI - Giving from our bodily belongings: is donation an appropriate paradigm for the giving of bodies and body parts? : What else might be considered? PMID- 19543691 TI - The major inflammatory mediator interleukin-6 and obesity. AB - Adipose tissue is one of the main sources of inflammatory mediators, with interleukin-6 (IL-6) among them. Although high systemic levels of inflammatory mediators are cachectogenic and/or anorexic, today it is a widely propagated thesis that in the background of obesity, a low level of chronic inflammation can be found, with IL-6 being one of the many suggested mediators. This paper reviews the studies describing elevated IL-6 levels in obese patients and the role of adipocytes and adipose-tissue macrophages in the production of IL-6. The secretion of IL-6 is regulated by several physiologic or pathologic factors: hormones, cytokines, diet, physical activity, stress, hypoxia, and others. Adipose tissue-derived IL-6 may have an effect on metabolism through several mechanisms, including adipose tissue-specific gene expression, triglyceride release, lipoprotein lipase downregulation, insulin sensitivity, and so on. Having a better understanding of these mechanisms may contribute to the prevention and treatment of obesity. PMID- 19543694 TI - Influence of magnetic field on activity of given anaerobic sludge. AB - Two modes of magnetic fields were applied in the Cr(6+) removal sludge reactors containing two predominated strains--Bacillus sp. and Brevibacillus sp., respectively. The magnetic field mode I* of 0-4.5 or 0-14 mT between pieces was obtained by setting the magnetic pieces with the surface magnetic density of 0-6 or 0-20 mT into the reactor, and the magnetic field mode II* of 6, 20, or 40 mT on the return line was obtained by controlling the working distance of the permanent magnet outside the sludge return line. The effects of different magnetic fields on the activity of the given anaerobic sludge were studied by comparing with the control (absent of magnetic field). The results showed that the magnetic field of 0-4 mT improved the activity of given sludge most effectively, U(max) CH(4) (the peak methane-producing rate) and the methane producing volume per gCOD(Cr) reached 64.3 mlCH(4)/gVSS.d and 124 mlCH(4)/gCOD(Cr), which increased by 20.6 and 70.7%, respectively, compared with the control. And the magnetic field of 20 mT took second place. It could be concluded that the input of some magnetic field could improve the activity of anaerobic sludge by increasing the transformation efficiency of COD(Cr) matters to methane, and the total organic wastage did not increase. PMID- 19543695 TI - Plasma salicylate level and aspirin resistance in survivors of myocardial infarction. AB - To investigate the effect of aspirin on the platelets of survivors of myocardial infarction we correlated plasma salicylate level with platelet reactivity in ten patients and ten normal controls. The patients and controls were tested at the end of 2 week periods on 75, 150 and 300 mg aspirin daily by mouth. Platelet reactivity was measured, under high shear stress conditions, using cartridges containing adrenaline and adenosine diphosphate in a PFA-100 platelet function analyser. The time taken by the developing platelet aggregate to close an aperture in the collagen membrane of the cartridge, the closure time, was taken as an index of platelet reactivity. There was no difference in baseline haematocrit, platelet count or plasma vWF antigen level between the groups. There was a dose-dependent increase in closure time of the adrenaline containing cartridge in the controls (P < 0.001), but not in the patients (P = 0.08), compatible with a reduced anti-platelet effect of aspirin in the patients. Furthermore, plasma salicylate level was higher in the patient group (P < 0.05). PMID- 19543697 TI - Associations of serum Ca and Mg levels with mental health in adult women without psychiatric disorders. AB - Several lines of evidence from previous studies suggest that Calcium (Ca) and Magnesium (Mg) may be involved in intracellular and interneuronal processes associated with affective disorders. However, there have been inconsistent results on the effect of Ca and Mg on depressive mood disorder. This cross sectional study was conducted to determine whether serum Ca and Mg levels, as well as serum Ca/Mg ratio, are associated with mental health in relatively healthy, adult women without psychiatric disorders. One hundred and twelve adult women were recruited from the outpatient clinic in a university hospital setting. Serum Ca and Mg levels were measured and indicators of mental health such as depression, anxiety, and stress were evaluated using two validated questionnaires; the Hospital Anxiety Depression Scale and the Modified Brief Encounter Psychosocial Instrument Stress Scale. After categorizing the serum Ca and Mg levels, and the Ca/Mg ratio into tertiles, the mean scores on each mental health scale were compared using analysis of covariance. The risk of depressive mood disorder according to the tertiles of serum Mg level and serum Ca/Mg ratio was assessed using logistic regression analysis. Women in the middle tertile of serum Ca/Mg ratio had significantly lower scores on depression and stress scales (p = 0.004 and p = 0.007, respectively) and a lower odds ratio (OR) for the risk of depressive mood disorder (OR = 0.31, CI(95%) 0.10-0.93) than those in the highest tertile. The OR for the risk of depressive mood disorder was higher in women in the lowest tertile of serum Mg than in those in the highest tertile (OR = 3.92, CI(95%) 1.11-13.83). Serum Mg level and serum Ca/Mg ratio may be involved in the mechanism for the progression of depressive mood or stress perception in relatively healthy, adult women. PMID- 19543696 TI - A rationally designed heparin, M118, has anticoagulant activity similar to unfractionated heparin and different from Lovenox in a cell-based model of thrombin generation. AB - Unfractionated heparin (UFH) enhances antithrombin (AT) inhibition of thrombin (IIa) and factor Xa (FXa). Low molecular weight heparins (LMWH) primarily enhance AT inhibition of FXa. M118 is a LMWH produced from UFH and retains its ability to promote both FXa and IIa inhibition. We tested the hypothesis that M118 has anticoagulant activities similar to UFH in an in vitro model of coagulation. Platelet IIa generation was assessed in a cell-based model that mimics aspects of coagulation in vivo. Inhibition of IIa generation as a function of concentration was steeper for UFH than Lovenox. The effect of M118 closely paralleled that of UFH. By contrast, M118 did not prolong the aPTT to as great a degree as UFH, though both prolonged the aPTT more than did Lovenox. Our data suggest that the ability to inhibit platelet surface IIa generation correlates with the therapeutic level of heparins and confirms similarities between the anticoagulant properties of M118 and UFH. PMID- 19543699 TI - [Cyberstalking]. AB - The term cyberstalking appears in the media with increasing frequency. So far epidemiological studies are sparse. Since researchers have used different definitions and study samples for cyberstalking, widely varying prevalence rates have been published. We report here a case of cyberstalking and discuss available empirical data. Cyberstalking may cause psychological distress similar to that of real world stalking. The need for a scientific definition of cyberstalking and for future studies is presented. Since it is likely that psychiatrists will encounter victims of cyberstalking they should have knowledge of this phenomenon. PMID- 19543698 TI - [Skin and teeth]. AB - Skin and teeth not only share several characteristics in their development but also have a similar ontogenetic origin from the ectoderm. Thus, numerous congenital diseases, such as genetic, infectious, inflammatory, and immune disorders affect both skin and teeth. Particularly skin diseases are often identified by the synopsis of systemic symptoms. The understanding and appreciation of dental abnormalities guides dermatologists in the diagnosis and identification of syndromes as well as in finding an optimal therapy. PMID- 19543700 TI - Health risk estimation due to carbon monoxide pollution at different spatial levels in Santiago, Chile. AB - The Metropolitan Region of Santiago has been declared as nonattainment area for carbon monoxide (CO). The data was based on seven air monitoring stations that do not necessarily represent the real population exposure. Usually, a series of coefficients that relates the concentration of pollutants with health effects (betas of the concentration-response equations) are used. For the calculation of these betas, normally, a city average air pollution concentration is used; nevertheless, these betas do not necessarily represent the real risk of exposure that the inhabitants are experimented. For this reason, this study estimates two aspects that are unknown and that are necessary to policymakers in public health decisions. First, estimate the exposure at a lower spatial resolution level and, second, estimate a concentration-response curve that differs spatially and depends on the proportion of susceptible groups. We followed the Air Pollution and Health, European Approach/Spanish Multicenter Study on the Relationship Between Air Pollution and the Mortality methodology for the determination of the short-term health effect due to CO population exposure at the three spatial levels (city, zone, and county). We found that there are different risks in cardiorespiratory mortality in the group over 64 years old associated to an unequal exposure to CO in Santiago city. The higher risks are in counties located to the south-east of the city. These finding confirm the necessity to face, in a different way, the approach when evaluating control strategies and policy decision to control air pollution in Santiago. PMID- 19543701 TI - Mayfly (Insecta: Ephemeroptera) community structure as an indicator of the ecological status of a stream in the Niger Delta area of Nigeria. AB - Ephemeroptera is an important group of insects used in the bioassessment and monitoring of freshwater bodies worldwide because of their relative abundance in a wide variety of substrates and their increasing chances of detecting pollution impacts. In this study, their faunistic composition and spatiotemporal variations in density and diversity in River Orogodo (Southern Nigeria) was investigated at five ecologically distinct stations over a 12-month period. The mayfly nymph community responses to environmental variables were evaluated by means of biological measures and multivariate analysis (redundancy analysis [RDA]). Thirteen morphologically distinct taxa belonging to six families were identified. The dominant taxa were Afrobaetodes pusillus (23.1%), Baetis sp. (13.7%), and Caenis cibaria (11.4%). The density of Ephemeroptera differed significantly (p < 0.05) both in space and time. Diversity was influenced by substrate heterogeneity which in turn was influenced by catchment processes such as flooding and anthropogenic activities especially abattoir effluent. Based on the RDA ordination and relative abundance data, Baetis sp. dominated at impacted stations while a more equitable distribution of species were observed in less disturbed sites. Water velocity, canopy cover, nature of bottom sediments, and the amount of dissolved oxygen also accounted for the variations in Ephemeroptera densities at the different stations. Shannon diversity, taxa richness, and evenness were lowest in station 3 (the abattoir discharge site). PMID- 19543702 TI - Whole-cell bioreporters for the detection of bioavailable metals. AB - Whole-cell bioreporters are living microorganisms that produce a specific, quantifiable output in response to target chemicals. Typically, whole-cell bioreporters combine a sensor element for the substance of interest and a reporter element coding for an easily detectable protein. The sensor element is responsible for recognizing the presence of an analyte. In the case of metal bioreporters, the sensor element consists of a DNA promoter region for a metal binding transcription factor fused to a promoterless reporter gene that encodes a signal-producing protein. In this review, we provide an overview of specific whole-cell bioreporters for heavy metals. Because the sensing of metals by bioreporter microorganisms is usually based on heavy metal resistance/homeostasis mechanisms, the basis of these mechanisms will also be discussed. The goal here is not to present a comprehensive summary of individual metal-specific bioreporters that have been constructed, but rather to express views on the theory and applications of metal-specific bioreporters and identify some directions for future research and development. PMID- 19543703 TI - Cell-based genotoxicity testing : genetically modified and genetically engineered bacteria in environmental genotoxicology. AB - Genotoxicity test systems that are based on bacteria display an important role in the detection and assessment of DNA damaging chemicals. They belong to the basic line of test systems due to their easy realization, rapidness, broad applicability, high sensitivity and good reproducibility. Since the development of the Salmonella microsomal mutagenicity assay by Ames and coworkers in the early 1970s, significant development in bacterial genotoxicity assays was achieved and is still a subject matter of research. The basic principle of the mutagenicity assay is a reversion of a growth inhibited bacterial strain, e.g., due to auxotrophy, back to a fast growing phenotype (regain of prototrophy). Deeper knowledge of the -mutation events allows a mechanistic understanding of the induced DNA-damage by the utilization of base specific tester strains. Collections of such specific tester strains were extended by genetic engineering. Beside the reversion assays, test systems utilizing the bacterial SOS-response were invented. These methods are based on the fusion of various SOS-responsive promoters with a broad variety of reporter genes facilitating numerous methods of signal detection. A very important aspect of genotoxicity testing is the bioactivation of -xenobiotics to DNA-damaging compounds. Most widely used is the extracellular metabolic activation by making use of rodent liver homogenates. Again, genetic engineering allows the construction of highly sophisticated bacterial tester strains with significantly enhanced sensitivity due to overexpression of enzymes that are involved in the metabolism of xenobiotics. This provides mechanistic insights into the toxification and detoxification pathways of xenobiotics and helps explaining the chemical nature of hazardous substances in unknown mixtures. In summary, beginning with "natural" tester strains the rational design of bacteria led to highly specific and sensitive tools for a rapid, reliable and cost effective -genotoxicity testing that is of outstanding importance in the risk assessment of compounds (REACH) and in ecotoxicology. PMID- 19543704 TI - Monitoring mammalian cell cultivations for monoclonal antibody production using near-infrared spectroscopy. AB - Near-infrared (NIR) spectroscopy as a process monitoring and process supervision technique is reviewed in the context of biomanufacturing.An industrial pilot plant mammalian cell cultivation process has been chosen to illustrate the use of on-line in-situ NIR monitoring by means of an immersion transflectance NIR probe.NIR calibration development must be performed carefully and should incorporate a number of steps to obtain a properly validated model which exhibits long-term robustness and is independent of process scale. A description of such good modelling practises is given. In general, NIR can be as accurate as the reference methods employed and at least as precise provided that sufficient spectral selectivity and sensitivity exists.NIR can also be used as a direct technique for very fast process monitoring and process supervision, thus enabling one to follow the trajectory of a process. This alternative to the indirect use of NIR through laborious calibration development with direct reference methods has been little explored. Since NIR is sensitive to both chemical and physical properties, the analysis of whole samples enables relevant process information to be captured and thus generates better process state estimates than by simply looking at defined process parameters one at a time. PMID- 19543705 TI - On-chip detection of cellular activity. AB - The use of on-chip cellular activity monitoring for biological/chemical sensing is promising for environmental, medical and pharmaceutical applications. The miniaturization revolution in microelectronics is harnessed to provide on-chip detection of cellular activity, opening new horizons for miniature, fast, low cost and portable screening and monitoring devices. In this chapter we survey different on-chip cellular activity detection technologies based on electrochemical, bio-impedance and optical detection. Both prokaryotic and eukaryotic cell-on-chip technologies are mentioned and reviewed. PMID- 19543706 TI - Stem cells for cardiac regeneration by cell therapy and myocardial tissue engineering. AB - Congestive heart failure, which often occurs progressively following a myocardial infarction, is characterized by impaired myocardial perfusion, ventricular dilatation, and cardiac dysfunction. Novel treatments are required to reverse these effects - especially in older patients whose endogenous regenerative responses to currently available therapies are limited by age. This review explores the current state of research for two related approaches to cardiac regeneration: cell therapy and tissue engineering. First, to evaluate cell therapy, we review the effectiveness of various cell types for their ability to limit ventricular dilatation and promote functional recovery following implantation into a damaged heart. Next, to assess tissue engineering, we discuss the characteristics of several biomaterials for their potential to physically support the infarcted myocardium and promote implanted cell survival following cardiac injury. Finally, looking ahead, we present recent findings suggesting that hybrid constructs combining a biomaterial with stem and supporting cells may be the most effective approaches to cardiac regeneration. PMID- 19543708 TI - Soluble triggering receptor expressed on myeloid cells-1 and diagnosis of ventilator-associated pneumonia. PMID- 19543709 TI - Research ethics committees in Europe: trials and tribulations. AB - INTRODUCTION: Ethics committees have been an integral part of clinical research since 1975, when they were introduced through the amendment of the Declaration of Helsinki. Every proposal for clinical research on human subjects has to be submitted to an independent ethics committee for review and approval. The European Clinical Trials Directive 2001/20/EC was implemented in 2004 to harmonise the legislative framework for clinical research in Europe in order to make Europe more competitive in clinical research while at the same time improving the protection of research participants. RESULTS: We have evaluated the situation of ethics committees in Europe five years after the implementation of the new law with special consideration of the number of Ethics Committees per European Member State and the number of members within the specific committees, including the selection of members, also in regard to gender aspects and training requirements, the remuneration or compensation of members in regard of their review obligations, and also issues of conflicts of interest. CONCLUSION: Inadequate remuneration for professional services and gender imbalance are universal concerns across Europe. As the position of ethics committees changes continuously towards greater responsibility, further guidance is needed to uniformly adapt their structures to those needs. PMID- 19543710 TI - Characterization of Bacillus subtilis YfkE (ChaA): a calcium-specific Ca2+/H+ antiporter of the CaCA family. AB - YfkE, a protein from Bacillus subtilis, exhibits homology to the Ca(2+):Cation Antiporter (CaCA) Family. In a fluorescence-based assay of everted membrane vesicles prepared from Na(+)(Ca(2+))/H(+) antiporter-defective mutant Escherichia coli KNabc, YfkE exhibited robust Ca(2+)/H(+) antiport activity, with a K (m) for Ca(2+) estimated at 12.5 muM at pH 8.5 and 113 muM at pH 7.5. Neither Na(+) nor K(+) served as a substrate. Mg(2+) also did not serve as a substrate, but inhibited the Ca(2+)/H(+) antiporter activity. The Ca(2+) transport capability of YfkE was also observed directly by transport assays in everted membrane vesicles using radiolabeled (45)Ca(2+). Transcriptional analysis from the putative yfkED operon using beta-garactosidase activity as a reporter revealed that both of the yfkE and yfkD genes are regulated by forespore-specific sigma factor, SigG, and the general stress response regulator, SigB. These results suggest that YfkE may be needed for Ca(2+) signaling in the sporulation or germination process in B. subtilis. ChaA is proposed as the designation for YfkE of B. subtilis. PMID- 19543711 TI - Comment on Slama R, Cyrys J, Herbarth O, Wichmann H-E, Heinrich J. saying: "The authors did not wish to reply, given Dr. Morfeld's persistence in refusing to fill in the conflict of interest statement and in misleadingly quoting parts of the sentences of our publications". PMID- 19543712 TI - Occupational exposure to polycyclic aromatic hydrocarbons and DNA damage by industry: a nationwide study in Germany. AB - Exposure to polycyclic aromatic hydrocarbons (PAH) and DNA damage were analyzed in coke oven (n = 37), refractory (n = 96), graphite electrode (n = 26), and converter workers (n = 12), whereas construction workers (n = 48) served as referents. PAH exposure was assessed by personal air sampling during shift and biological monitoring in urine post shift (1-hydroxypyrene, 1-OHP and 1-, 2 + 9-, 3-, 4-hydroxyphenanthrenes, SigmaOHPHE). DNA damage was measured by 8-oxo-7,8 dihydro-2'-deoxyguanosine (8-oxodGuo) and DNA strand breaks in blood post shift. Median 1-OHP and SigmaOHPHE were highest in converter workers (13.5 and 37.2 microg/g crea). The industrial setting contributed to the metabolite concentrations rather than the air-borne concentration alone. Other routes of uptake, probably dermal, influenced associations between air-borne concentrations and levels of PAH metabolites in urine making biomonitoring results preferred parameters to assess exposure to PAH. DNA damage in terms of 8-oxo-dGuo and DNA strand breaks was higher in exposed workers compared to referents ranking highest for graphite-electrode production. The type of industry contributed to genotoxic DNA damage and DNA damage was not unequivocally associated to PAH on the individual level most likely due to potential contributions of co-exposures. PMID- 19543714 TI - Use of 18O water and ESI-MS detection in subsite characterisation and investigation of the hydrolytic action of an endoglucanase. AB - We present a novel method for investigating subsite-substrate interactions of glycoside hydrolases and the determination of the oligosaccharide cleavage point based on the analysis of the hydrolysis products produced in the presence of (18)O-labelled water. Conventional techniques for such determination of the hydrolysis pattern call for the chemical modification of the substrate, whereas the method presented makes it possible to use natural substrates, utilising the selectivity and sensitivity of mass spectrometry. This method is very useful for the detection and analysis of enzyme-catalysed hydrolysis, provided that the conditions are chosen where (18)O incorporation without the presence of the enzyme is absent or undetectable. Such conditions were found and used in incubations of cellopentaose with the well-characterised endoglucanase Cel5A from Bacillus agaradhaerens. We were able to confirm that the preferred glycoside bond to be hydrolysed is the third one counting from the non-reducing end of the cellopentaose. Thus, cellopentaose prefers to bind from the -3 to the +2 subsites, which is in accordance with published crystallographic data. The main advantage of the method presented is that there is no need for a priori chemical modification/labelling of oligosaccharide substrates, which are processes that can disturb the enzyme-substrate interaction. From (18)O incorporation we could demonstrate that the enzyme also has an oxygen-exchange activity on cellotriose and cellobiose. This is in agreement with the mechanism for transglycosylation and indicates that it is possible for the enzyme to perform such reactions. PMID- 19543715 TI - A microsystem of low-voltage-driven electrophoresis on microchip with array electrode pairs for the separation of amino acids. AB - In this paper, a new approach for the separation of amino acids on the electrophoresis chip-based low-voltage-driven electrophoresis was reported in detail. This low-voltage-driven electrophoresis process could be realized by powering directly the arrayed electrode pairs with low direct current (DC) voltage to generate a moving electric field along the separation microchannel, which could maintain enough electric field strength for electrophoresis. The proposed microfluidic electrophoresis chip was bonded directly with silicon-on insulator (SOI) substrate and polydimethylsiloxane (PDMS) cover plate at room temperature. The microfluidic channels and the arrayed electrodes were etched on SOI wafer by silicon microelectromechanical system technology. A specially integrated circuit was proposed to power a 30-60-V DC voltage to particular sets of these electrode pairs in a controlled sequence such that the moving electric field could be formed, and the low-voltage-driven electrophoresis could be realized in the microchannel. In the experiments, with 10(-4) mol/L phenylalanine and lysine as analytes, the separation of amino acids on the low-voltage-driven electrophoresis microchip was conducted by homemade integrated control circuit; a method for separating amino acids was well established. It was also shown that the phenylalanine and lysine mixture was effectively separated in less than 7 min and with a resolution of 2.0. To the best of our knowledge, the low-voltage driven microchip electrophoresis device could be of potential prospective in the fields of integrated and miniaturized biochemical analysis system. PMID- 19543716 TI - Forensic analysis of anthraquinone, azo, and metal complex acid dyes from nylon fibers by micro-extraction and capillary electrophoresis. AB - The extraction and separation of dyes present on textile fibers offers the possibility of enhanced discrimination between forensic trace fiber evidence. An automated liquid sample handling workstation was programmed to deliver varying solvent combinations to acid-dyed nylon samples, and the resulting extracts were analyzed by an ultraviolet/visible microplate reader to evaluate extraction efficiencies at different experimental conditions. Combinatorial experiments using three-component mixture designs varied three solvents (water, pyridine, and aqueous ammonia) and were employed at different extraction temperatures for various extraction durations. The extraction efficiency as a function of the three solvents (pyridine/ammonia/water) was modeled and used to define optimum conditions for the extraction of three subclasses of acid dyes (anthraquinone, azo, and metal complex) from nylon fibers. The capillary electrophoresis analysis of acid dye extracts is demonstrated using an electrolyte solution of 15 mM ammonium acetate in acetonitrile/water (40:60, v/v) at pH 9.3. Excellent separations and discriminating diode array spectra are obtained even for dyes of similar color. PMID- 19543717 TI - Multivariate standard addition method solved by net analyte signal calculation and rank annihilation factor analysis. AB - This article describes the use of the net analyte signal (NAS) concept and rank annihilation factor analysis (RAFA) for building two different multivariate standard addition models called "SANAS" and "SARAF." In the former, by the definition of a new subspace, the NAS vector of the analyte of interest in an unknown sample as well as the NAS vectors of samples spiked with various amounts of the standard solutions are calculated and then their Euclidean norms are plotted against the concentration of added standard. In this way, a simple linear standard addition graph similar to that in univariate calibration is obtained, from which the concentration of the analyte in the unknown sample and the analytical figures of merit are readily calculated. In the SARAF method, the concentration of the analyte in the unknown sample is varied iteratively until the contribution of the analyte in the response data matrix is completely annihilated. The proposed methods were evaluated by analyzing simulated absorbance data as well as by the analysis of two indicators in synthetic matrices as experimental data. The resultant predicted concentrations of unknown samples showed that the SANAS and SARAF methods both produced accurate results with relative errors of prediction lower than 5% in most cases. PMID- 19543718 TI - Quantitative methods for food allergens: a review. AB - The quantitative detection of allergens in the food chain is a strategic health objective as the prevalence of allergy continues to rise. Food allergenicity is caused by proteins either in their native form or in forms resulting from food processing. Progress in mass spectrometry greatly opened up the field of proteomics. These advances are now available for the detection and the quantification of traces of allergenic proteins in complex mixtures, and complete the set of biological tests used until now, such as ELISA or PCR. We review methods classified according to their ability to simultaneously quantify and identify allergenic proteins and underline major advances in the mass spectrometric methods. PMID- 19543719 TI - Multiplexing with SERS labels using mixed SAMs of Raman reporter molecules. AB - Surface-enhanced Raman scattering (SERS) offers a tremendous multiplexing capacity for the selective detection of biomolecules in targeted research. SERS labels comprising self-assembled monolayers (SAMs) of Raman reporter molecules on the surface of metal nanoparticles are sensitive and robust probes. Advantages of a SAM include maximum sensitivity, minimal unwanted co-adsorption of molecules from the surroundings, and reproducible SERS spectra with only few dominant Raman bands--all of these independent of a particular SERS substrate. We demonstrate experimentally how to increase the multiplexing capacity of SERS labels by using mixed SAMs with up to three different Raman reporter molecules on the surface of the metal colloid. Type and stoichiometry of a particular Raman label in a multi component SAM are additional parameters compared with one-component SAMs. All one , two-, and three-component SAMs on gold nanospheres can be easily discriminated, either by their original SERS spectra or the corresponding bar codes. PMID- 19543720 TI - Learning and generalization of time production in humans: rules of transfer across modalities and interval durations. AB - This article investigated both the ability of naive human subjects to learn interval production, as well as the properties of learning generalization across modalities and interval durations that varied systematically from the over trained interval. Human subjects trained on a 450-, 650-, or 850-ms single interval production task, using auditory stimuli to define the intervals, showed a significant decrease in performance variability with intensive training. This learning generalized to the visual modality and to non-trained durations following a Gaussian transfer pattern. However, the learning carryover followed different rules, depending on the duration of the trained interval as follows: (1) the dispersion of the generalization curve increased as a function of the trained interval, (2) the generalization pattern was tilted to the right in the visual condition, and (3) the transfer magnitude for 650 ms was less prominent than for the other two intervals. These findings suggest the existence of neural circuits that are tuned to specific time lengths and that show different temporal processing properties depending on their preferred interval duration. PMID- 19543721 TI - TMS disruption of V5/MT+ indicates a role for the dorsal stream in word recognition. AB - Although word recognition is a skill commonly expected to rely more on ventral rather than dorsal stream processing, there is some evidence for a magnocellular/dorsal impairment in dyslexia. The early rapid feedforward/feedback loop through the dorsal stream seen in primate has been suggested to allow an initial global analysis, and in human early activation of parietal attention mechanisms for detecting salient stimuli, facilitating more local level detailed ventral stream processing. To test this model in humans, transcranial magnetic stimulation (TMS) was used to probe the role of early visual cortex (V1/V2) and V5/MT+ in single word identification. TMS over V1/V2 between word onset and 36 ms post word onset disrupted accurate word discrimination, with disruption also evident at approximately 99 ms. TMS over V5/MT+ also disrupted accuracy following stimulation at approximately the same time as word onset and again at 130 ms post word onset. Thus, a role for V5/MT+ in accurate single word identification is apparent suggesting rapid triggering of attention to salient exogenous stimuli may be required prior to processing in primary and temporal cortical regions. PMID- 19543722 TI - Visuomotor transformation for interception: catching while fixating. AB - Catching a ball involves a dynamic transformation of visual information about ball motion into motor commands for moving the hand to the right place at the right time. We previously formulated a neural model for this transformation to account for the consistent leftward movement biases observed in our catching experiments. According to the model, these biases arise within the representation of target motion as well as within the transformation from a gaze-centered to a body-centered movement command. Here, we examine the validity of the latter aspect of our model in a catching task involving gaze fixation. Gaze fixation should systematically influence biases in catching movements, because in the model movement commands are only generated in the direction perpendicular to the gaze direction. Twelve participants caught balls while gazing at a fixation point positioned either straight ahead or 14 degrees to the right. Four participants were excluded because they could not adequately maintain fixation. We again observed a consistent leftward movement bias, but the catching movements were unaffected by fixation direction. This result refutes our proposal that the leftward bias partly arises within the visuomotor transformation, and suggests instead that the bias predominantly arises within the early representation of target motion, specifically through an imbalance in the represented radial and azimuthal target motion. PMID- 19543723 TI - Analysis of protein mobilities and interactions in living cells by multifocal fluorescence fluctuation microscopy. AB - The spatial and temporal fluctuation microscope (STFM) presented here extends the concept of a fluorescence confocal laser scanning microscope to illumination and detection along a line. The parallel multichannel acquisition of the fluorescence signal was accomplished by using a single line of an electron-multiplying charge coupled device camera at 14 mus time resolution for detection of the fluorescence signal. The STFM system provided fast confocal imaging (30 images per second) and allowed for the spatially resolved detection of particle concentration fluctuations in fluorescence correlation spectroscopy experiments. For the application of the STFM, an approximated theoretical description of the beam geometry, the point-spread function, and the fluorescence auto- and cross correlation functions were derived. The STFM was applied to studies of the dynamics of promyelocytic leukemia nuclear bodies, green fluorescent protein, and chromatin-remodeling complexes in living cells. The results demonstrate the unique capabilities of the STFM for characterizing the position-dependent translocations and interactions of proteins in the cell. PMID- 19543725 TI - The MR dimple sign in irreducible posterolateral knee dislocations. AB - While posterolateral knee dislocation is a rare entity, its identification has important implications. The invagination of skin seen in posterolateral knee dislocations is referred to as the "dimple sign" on physical exam. We report a case where the "dimple sign" was also demonstrated on magnetic resonance imaging. Failure to recognize this finding on imaging will render the knee irreducible, as a result of interposed tissues, until discovered by the orthopedic surgeon intraoperatively. The incarcerated medial capsule and medial collateral ligament could also be misinterpreted as torn meniscus, as occurred in our case. PMID- 19543724 TI - Long-term performance and microbial community analysis of a full-scale synthesis gas fed reactor treating sulfate- and zinc-rich wastewater. AB - The performance of a full-scale (500 m3) sulfidogenic synthesis gas fed gas-lift reactor treating metal- and sulfate-rich wastewater was investigated over a period of 128 weeks. After startup, the reactor had a high methanogenic activity of 46 Nm3.h(-1). Lowering the carbon dioxide feed rate during the first 6 weeks gradually lowered the methane production rate. Between weeks 8 and 93, less than 1% of the hydrogen supplied was used for methanogenesis. Denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified 16S rRNA gene fragments showed that the archaeal community decreased in diversity but did not disappear completely. After the carbon dioxide feed rate increased in week 88, the methane production rate also increased, confirming that methane production was carbon dioxide limited. Even though lowering the carbon dioxide feed appeared to affect part of the sulfate-reducing community, it did not prevent achieving the desired rates of sulfate reduction. The average sulfate conversion rate was 181 kg.h(-1) for the first 92 weeks. After 92 weeks, the sulfate input rate was increased and from week 94 to 128, the average weekly sulfate conversion rate was 295 kg.h(-1) (SD+/-87). Even higher sulfate conversion rates of up to 400 kg.h( 1) could be sustained for weeks 120-128. The long-term performance and stability together with the ability to control methanogenesis demonstrates that synthesis gas fed reactor can be used successfully at full scale to treat metal and sulfate rich wastewater. PMID- 19543726 TI - Construction of a DNA vaccine encoding Flk-1 extracellular domain and C3d fusion gene and investigation of its suppressing effect on tumor growth. AB - Although the critical role of complement component C3d as a molecular adjuvant in preventing virus infection is well established, its role in cancer prophylaxis and treatment is unclear. In this study, we constructed a recombinant plasmid encoding Flk-1 and C3d3 fusion proteins and investigated its transient expression in vitro in transfected eukaryotic cells and its antibody response in immunized mice. Subsequently, we investigated the vaccine's ability to elicit an immune response leading to suppression of angiogenesis and tumor growth in mice bearing bladder transitional cell carcinoma. Using Western blotting, immunocytochemistry, and flow cytometry, we detected the expression of Flk-1 and C3d3 fusion proteins in COS-7 cells transfected with these recombinant plasmids. Further binding experiment using CR2 (C3d receptor) positive Raji cells that were incubated with transfected COS-7 supernatant indicated that C3d was successfully fused to Flk-1. Although both vaccines elicited peak antibody levels at 5 weeks, Flk-1-specific antibody titer in pSG.SS.Flk-1(ECD).C3d3.YL-immunized mice was significantly higher when compared to pSG.SS.Flk-1(ECD).YL-immunized mice. The results of experiments with bladder tumor-bearing mice showed that the vaccine inhibited tumor growth significantly. These results suggest that C3d plays a critical role in tumor immunotherapy by promoting antibody response in Flk-1-based DNA vaccines. This approach may provide a new strategy for the rational design of anti-angiogenic therapies for the treatment of solid tumors and provide a basis for the further exploitation and application of the anti-angiogenesis DNA vaccines. PMID- 19543727 TI - Comparison of various criteria in predicting treatment response and prognosis of patients with myelodysplastic syndrome treated with azacitidine. AB - This study was performed to identify whether cytogenetics, International Prognostic Scoring System (IPSS), or World Health Organization Classification Based Prognostic Scoring System are predictive of the efficacy of azacitidine in patients with myelodysplastic syndrome (MDS). We retrospectively reviewed the clinical records of 113 patients with MDS treated with azacitidine. The "response alternating disease natural history," "cytogenetic response," and "hematologic improvement" were assessed by serial bone marrow biopsy, cytogenetic study, and hemogram analyses. The complete and partial remission rates were 17.6% and 3.9% in 51 evaluable patients. There were no significant differences in response rate in the different cytogenetic/IPSS/WPSS groups. The overall hematologic response (HR) rate was 49.6%, and the HR rate was significantly greater in patients classed as "very high" risk according to the WPSS compared with other patient groups. The 1-year overall survival (OS) rate was higher among patients with HR compared with those without HR (80.9% vs 63.3%, p = 0.046), and the 1-year OS rate among patients classed as being at high risk by each criteria was similar to that of patients classed as being at low risk. The hazard ratio of death among patients with HR compared with those without HR was 0.17 (95% CI 0.04-0.69) for high + very high risk group based on WPSS. Patients in the WPSS high-risk group had an increased HR rate compared with other patient groups, and the achievement of HR was associated with a significant increase in OS. Azacitidine showed similar efficacy in all patient groups, even in patients with poor cytogenetics and in high-risk groups. PMID- 19543728 TI - Temozolomide in malignant gliomas: current use and future targets. AB - Temozolomide (TMZ) is an oral alkylating agent that is regarded as a tolerable and effective drug. When combined with radiotherapy in patients with newly diagnosed glioblastoma, survival is significantly prolonged. This finding has led to widespread use of TMZ for patients with this disease. We summarize developing concerns regarding the use of TMZ, imaging of malignant gliomas, and the pharmacology of TMZ-mechanism of action, scheduling and strategies for overcoming resistance. PMID- 19543729 TI - Gene expression analysis of drug-resistant MCF-7 cells: implications for relation to extracellular matrix proteins. AB - PURPOSE: Since multidrug resistance is a multifactorial phenomenon, a large-scale expression analysis of drug-resistant cells by using high-density oligonucleotide microarrays may provide information about new candidate genes contributing to resistance. Extracellular matrix (ECM) is responsible for many aspects of proliferation and invasive/metastatic behavior of tumor cells. This study demonstrates alterations in gene expression levels of several ECM components, matrix metalloproteinases (MMPs), adamalysins (ADAMs and ADAMTSs) and tissue inhibitors of metalloproteinases (TIMPs) in paclitaxel, docetaxel, vincristine and doxorubicin-resistant MCF-7 cells. METHODS: Resistant MCF-7 cells were developed by stepwise selection of cells in increasing concentrations of drugs. Affymetrix GeneChip Human Genome U133 Plus 2.0 Array was used for hybridizations. Statistical significance was determined by independent sample t test. The genes having altered expression levels in drug-resistant sublines were selected and filtered by volcano plots. RESULTS: Genes up/downregulated more than twofolds were selected and listed. Expression of 25 genes encoding ECM proteins (including collagen, finronectin and syndecan) and integrin receptor subunits were found to be upregulated in drug-resistant cells. In addition, expression levels of, 13 genes encoding MMPs, ADAMs, ADAMTSs and TIMPs (including MMP1, MMP9, ADAM9 and TIMP3) were found to be altered in drug-resistant sublines when compared with sensitive MCF-7. CONCLUSIONS: Based on the expression analysis profiles, this report provides a preliminary insight into the relationship between drug resistance and ECM components, which are related to invasion and metastasis. Correlation of each specific ECM component with drug resistance requires further analysis. PMID- 19543730 TI - Macrophagic myofascitis associated with rheumatoid arthritis. AB - Macrophagic myofascitis (MMF) is an unusual inflammatory myopathy characterized by muscle infiltration by macrophages and lymphocytes. Here, we describe a case of MMF which is associated with rheumatoid arthritis. A 53-year-old Japanese rheumatoid arthritis (RA) patient presented with focal tenderness of lower extremities. Magnetic resonance imaging showed evidence of myofascitis involving fascias of anterior tibialis muscle. Muscle biopsy showed a unique pathological pattern of MMF. MMF is known to be associated with vaccination containing aluminum. However, our case was not related to aluminum containing vaccinations and etiologies are unknown. The possible link needs to be discussed. PMID- 19543731 TI - Discoid lupus erythematosus in a patient with scleroderma and hepatitis C virus infection. AB - A 49-year-old Japanese woman presented with discoid lupus erythematosus (DLE) on the face. The presence of Raynaud's phenomenon, swollen fingers, a high anti nuclear antibody titer, and the results of a biopsy revealed limited-type systemic sclerosis (lSSc). The association of SSc with DLE is rare, although some single case reports have been published in Japan. Our patient was positive for hepatitis C virus infection. Racial predisposition and immune imbalance are proposed to have played a role in the development of these lesions in our case. PMID- 19543732 TI - Doubled-haploid production in chickpea (Cicer arietinum L.): role of stress treatments. AB - This is the first report on the production of double-haploid chickpea embryos and regenerated plants through anther culture using Canadian cultivar CDC Xena (kabuli) and Australian cultivar Sonali (desi). Maximum anther induction rates were 69% for Sonali and 63% for CDC Xena. Under optimal conditions, embryo formation occurred within 15-20 days of culture initiation with 2.3 embryos produced per anther for CDC Xena and 2.0 embryos per anther for Sonali. For anther induction, the following stress treatments were used: (1) flower clusters were treated at 4 degrees C for 4 days, (2) anthers were subjected to electric shock treatment of three exponentially decaying pulses of 50-400 V with 25 microF capacitance and 25 Omega resistance, (3) anthers were centrifuged at 168-1,509g for 2-15 min, and finally (4) anthers were cultured for 4 days in high-osmotic pressure (563 mmol) liquid medium. Anthers were then transferred to a solid embryo development medium and, 15-20 days later, embryo development was observed concomitant with a small amount of callus growth of 0.1-3 mm. Anther-derived embryos were regenerated on plant regeneration medium. Electroporation treatment of anthers enhanced root formation, which is often a major hurdle in legume regeneration protocols. Cytological studies using DAPI staining showed a wide range of ploidy levels from haploid to tetraploid in 10-30-day-old calli. Flow cytometric analysis of calli, embryos and regenerated plants showed haploid profiles and/or spontaneous doubling of the chromosomes during early regeneration stages. PMID- 19543734 TI - Minimally invasive vas surgery using a newly designed double-ringed clamp. AB - INTRODUCTION: We have recently designed a novel surgical instrument, a double ringed clamp (Moon's clamp), with which the vas deferens can be located and isolated both simply and safely while minimizing unnecessary injury to the surrounding tissue. MATERIALS AND METHODS: With the use of the Moon's clamp, we successfully performed 1,140 cases of simplified, minimally invasive vasectomy and 216 cases of mini-incision vasectomy reversals (vasovasostomies) without any significant complications. CONCLUSION: This surgical technique, using Moon's clamp, may be effective in reducing the operative time, the postoperative complications and the recovery time after different types of vas surgeries, such as vasectomy or vasectomy reversal. PMID- 19543733 TI - Brain ventricular wall movement assessed by a gated cine MR trueFISP sequence in patients treated with endoscopic third ventriculostomy. AB - The purpose of this study was to investigate the value of brain ventricular wall movement assessment with a gated cine trueFISP MR sequence for the diagnosis of endoscopic third ventriculostomy (ETV) patency. Sixteen healthy volunteers and ten consecutive patients with noncommunicating hydrocephalus were explored with a MR scanner (Siemens, Avanto 1.5 T) before, 1 week and 3 months after ETV. TrueFISP was evaluated qualitatively (ventricular wall movement and CSF flow through ETV) and quantitatively [distance moved (DMLT) during a cardiac cycle by the lamina terminalis]. The third ventricle volume (TVV) was assessed. Statistical analysis was performed using nonparametric tests. There was no motion of the lamina terminalis (LT) detected on preoperative data. A pulsatile motion of the LT was found for patients with a patent ETV and for controls. DMLT and TVV were correlated (r = 0.79, P = 0.006). A transient dysfunction of ETV was successfully diagnosed on the trueFISP sequence with no motion of the LT or CSF flow observed. The trueFISP sequence appears reliable for the diagnosis of ETV patency and provides non-invasive assessment of the movement of the ventricular wall related to CSF pressure changes. PMID- 19543735 TI - Comments on the article "Biomarkers for prostate cancer" by Eric Schiffer. PMID- 19543736 TI - [Keratoplasty: laminate or penetrate? Part 2: lamellar keratoplasty]. AB - A number of corneal disorders sooner or later call for a decision as to whether corneal grafting is necessary. In contrast to the current gold standard, penetrating keratoplasty (PKP), former lamellar techniques had the disadvantage of worse visual outcomes due to interface problems. A number of recent new technical developments have led to a 'renaissance' of various lamellar keratoplasty methods. Deep anterior lamellar keratoplasty (DALK) avoids the risk of endothelial immune reactions. However, visual outcomes are comparable to PKP only is complete baring of Descemet's membrane is achieved. In Descemet stripping (automated) endothelial keratoplasty [DS(A)EK], a variation of posterior lamellar keratoplasty, only the diseased endothelium is removed with the Descetmet membrane (descemetorhexis). The lamellar, endothelium-bearing graft can subsequently be implanted via a corneoscleral/corneal tunnel, whereby little astigmatism is induced. The main advantage of DS(A)EK compared to PKP is faster visual recovery with almost unaltered refraction, although visual acuity is often slightly lower. However, valid study results are still lacking for this new surgical technique to allow a long-term comparison of the new techniques with conventional penetrating keratoplasty. PMID- 19543738 TI - Metachronous multiple carcinomas arising at a colostomy site 18 years after abdominoperineal resection. PMID- 19543737 TI - [Keratoplasty: laminate or penetrate? Part 1: penetrating keratoplasty]. AB - A number of corneal disorders sooner or later call for a decision as to whether corneal grafting is necessary. The problems of the current gold standard treatment, conventional penetrating keratoplasty (PKP), have yet to be fully solved, namely accelerated postoperative endothelial cell loss, endothelial immune reactions and the induction of astigmatism, which depends among other things on the trephination technique used. The use of a hand trephine may lead to different cutting angles between donor and recipient, resulting in increased postoperative astigmatism. This technique is therefore now only used for certain indications. One major advantage of mechanically guided trephine systems, which are now considered standard, is the possibility of trephining the donor disc from the epithelial side to avoid donor oversizing, therefore reducing postoperative astigmatism. Even less astigmatism is induced in keratoconus patients using an Excimer laser for trephination, as this technique does not lead to corneal deformation during trephination. Furthermore, this non-contact method can also be used to trephine unstable corneas. Femtosecond lasers allow the creation not only of straight but also profiled trephination edges, leading to more stable wound healing and hence earlier suture removal. PMID- 19543740 TI - Combined molecular analysis of BRAF and IDH1 distinguishes pilocytic astrocytoma from diffuse astrocytoma. AB - Separation of pilocytic astrocytoma from diffuse astrocytomas frequently poses problems mostly related to small sample size. Precise classification and grading are essential due to different therapeutic strategies prompted by diagnoses of pilocytic astrocytoma WHO grade I, diffuse astrocytomas WHO grade II or anaplastic astrocytoma WHO grade III. Recently, genomic aberrations with a high specificity for distinct glioma entities have been described. Pilocytic astrocytomas carry a duplication at chromosome band 7q34 containing a BRAF KIAA1549 gene fusion in the majority of cases. IDH1 mutations are observed very frequently in adult astrocytomas and IDH2 mutations have been reported in some astrocytomas. We examined a series of 120 astrocytomas including 70 pilocytic astrocytomas WHO grade I and 50 diffuse astrocytomas WHO grade II for both, BRAF KIAA1549 fusion with a newly developed FISH assay and mutations in IDH1 and IDH2 by direct sequencing. Pilocytic astrocytomas contained the BRAF fusion in 49 cases (70%) but neither IDH1 nor IDH2 mutations. Astrocytomas WHO grade II exhibited IDH1 mutations in 38 cases (76%) but neither IDH2 mutations nor BRAF fusions. Thus, combined molecular analysis of BRAF and IDH1 is a sensitive and highly specific approach to separate pilocytic astrocytoma from diffuse astrocytoma. PMID- 19543742 TI - Personal experience with tinnitus retraining therapy. AB - We present the results of tinnitus retraining therapy (TRT) in a group of patients suffering from tinnitus and/or hyperacusia. Based on the scores from a specific questionnaire and the Tinnitus Handicap Inventory (THI), the patients were classified into five categories and began therapy according to Jastreboff's criteria. Depending on the individual case, therapy envisaged counselling sessions, ambient sound enrichment, sound generators and hearing aids. At the end of the 18-month period, therapeutic success was observed in 79% of the patients. The initial numerical values of the scale of the symptoms and the THI seem predictive of treatment outcome. The use of instruments (sound generators) increases the success rate, but the study also demonstrates the effectiveness of counselling and ambient sound enrichment. Failures mainly involved patients with hypacusia who refused to wear hearing aids, as this influenced the effectiveness of ambient sound enrichment and counselling. Paralleling the data in the literature, the results demonstrate the effectiveness of TRT, which cannot be attributed to a placebo effect given the extended duration of treatment. PMID- 19543741 TI - Treatment of displaced intra-articular calcaneal fractures by ligamentotaxis: current concepts' review. AB - INTRODUCTION: A large variety of therapeutic modalities for calcaneal fractures have been described in the literature. No single treatment modality for displaced intra-articular calcaneal fractures has proven superior over the other. This review describes and compares the different percutaneous distractional approaches for intra-articular calcaneal fractures. The history, technique, anatomical and fracture considerations, limitations and the results of different distractional approaches reported in the literature are reviewed. METHOD: Literature review on different percutaneous distractional approaches for displaced intra-articular calcaneal fractures. RESULTS: Eight studies in which application of a distraction technique was used for the treatment of calcaneal fractures were identified. Because of the use of different classification, techniques, and outcome scoring systems, a meta-analysis was not possible. A literature review reveals overall fair to poor result in 10-29% of patients. Ten up to 26% of patients are unable to return to work after percutaneous treatment of their fracture. A secondary arthrodesis has to be performed in 2-15% of the cases. Infectious complications occur in 2-15%. Some loss of reduction is reported in 4-67%. CONCLUSION: Percutaneous distractional reduction and fixation appears to be a safe technique with overall good results and an acceptable complication rate, compared with other treatment modalities for displaced intra-articular calcaneal fractures. A meta-analysis, based on Cochrane Library criteria is not possible, because of a lack of level 1 and 2 trials on this subject. PMID- 19543743 TI - Characteristics of ClC7 Cl- channels and their inhibition in mutant (G215R) associated with autosomal dominant osteopetrosis type II in native osteoclasts and hClcn7 gene-expressing cells. AB - ClC7 Cl(-) channels (Clcn7) are crucial for osteoclastic bone resorption and have heterozygous mutation in autosomal osteopetrosis type II (ADO II) patients. Although extracellular acidification is known to induce ClC7 Cl(-) currents in Clcn7-transfected oocytes, other characteristics of this acid-induced Cl(-) current, as well as the effects of mutant Clcn7 in ADO II, remain to be determined. The present study showed that extracellular acidification evoked outward Cl(-) currents in mouse osteoclasts. Expression of wild-type human Clcn7 in HEK293 cells also induced a significant increase in acid-activated Cl(-) currents. These acid-activated Cl(-) currents were independent of intracellular acidification and [Ca(2+)]( i ) increase. HEK293 cells with the Clcn7 mutation associated with ADO II at G215R did not display these Cl(-) currents. These results suggest that osteoclastic ClC7 Cl(-) channels are activated under extracellar acidification and suppressed in Clcn7 mutant associated with ADO II during bone resorption. PMID- 19543744 TI - Expression of Wnt gene family and frizzled receptors in head and neck squamous cell carcinomas. AB - Genes of the Wnt and Frizzled class, expressed in HNSCC tissue and cell lines, have an established role in cell morphogenesis and differentiation, and also they have oncogenic properties. We studied Wnt and Fz genes as potential tumor associated markers in HNSCC by qPCR. Expression levels of Wnt and Fz genes in 22 unique frozen samples from HNSCC were measured. We also assessed possible correlation between the expression levels obtained in cancer samples in relation to clinicopathologic outcome. Wnt-1 was not expressed in the majority of the HNSCC studied, whereas Wnt-5A was the most strongly expressed by the malignant tumors. Wnt-10B expression levels were related with higher grade of undifferentiation. Related to Fz genes, Fz-5 showed more expression levels in no affectation of regional lymph nodes. Kaplan-Meier survival analyses suggest a reduced time of survival for low and high expression of Wnt-7A and Fz-5 mRNA, respectively. qPCR demonstrated that HNSCC express Wnt and Fz members, and suggested that Wnt and Fz signaling is activated in HNSCC cells. PMID- 19543746 TI - Studies on collar spines of echinostomatid trematodes. AB - This review is mainly concerned with the methods used to study collar spines in echinostomatid trematodes. The salient studies concerned with light microscopy on live and fixed larval and adult echinostomes are reviewed. Similar information is also provided for echinostomatid collar spines examined by scanning electron microscopy and transmission electron miscroscopy. The limited information on specific histochemical studies on these spines is also reviewed. Lastly, an examination of the few papers that have considered structure and function of collar spines is given. PMID- 19543745 TI - Glioblastoma cells express functional cell membrane receptors activated by daily used medical drugs. AB - PURPOSE: Calcium ions are highly versatile spacial and temporal intracellular signals of non-excitable cells and have an important impact on nearly every aspect of cellular life controlling cell growth, metabolism, fluid secretion, information processing, transcription, apoptosis, and motility. Neurons and glia respond to stimuli, including neurotransmitters, neuromodulators, and hormones, which increase the intracellular calcium concentration. The function of intracellular calcium in gliomas is unknown. Lots of daily used drugs may act via receptors that can be linked to the intracellular calcium system and therefore could influence glioma biology. METHODS: Glioma cells were loaded with the calcium ion sensitive dye Fura 2-AM. Subsequently, cells were stimulated with 25 different medical drugs for 30 s. The increase of free intracellular calcium ions was measured and calculated by a microscope-camera-computer-unit. RESULTS: Except for the buffer solution HEPES that served as negative control and for the cortisol derivative dexamethasone, all other 24 tested drugs induced a rise of intracellular calcium ions. The cellular calcium responses were classified into seven functional groups. The tested substances activated several types of calcium channels and receptors. CONCLUSIONS: Our study impressively demonstrates that medical drugs are potent inducers of intracellular calcium signals. Totally unexpected, the results show a high amount of functional cellular receptors and channels on glioma cells, which could be responsible for certain biological effects like migration and cell growth. This calcium imaging study proves the usability of the calcium imaging as a screening system for functional receptors on human glioma cells. PMID- 19543747 TI - Penetration of cercariae into the living human skin: Schistosoma mansoni vs. Trichobilharzia szidati. AB - We studied the skin invasion of Schistosoma mansoni cercariae by placing gamma irradiated and nonirradiated cercariae onto the living human skin and timing the behavior of 53 individuals. The skin invasion of S. mansoni was less efficient compared to the bird schistosome Trichobilharzia szidati. S. mansoni cercariae crept longer on the skin after attachment until they started penetration movements (median of 43 s [range of 15 s-6.58 min]; T. szidati, median of 8 s [range of 0-80 s]). Subsequent to this longer exploratory phase, 74% penetrated into wrinkles (T. szidati 84%), 22% into the smooth skin surface (T. szidati 0%), and 4% into hair follicles (T. szidati 16%). The S. mansoni cercariae needed, on average, 6.58 min (range of 1.57-13.13 min) for full entry, while T. szidati needed 4.0 min (range of 1.38-13.34 min); the fastest S. mansoni cercaria entered the skin within 94 s, while T. szidati entered within 83 s. Sixty percent of the S. mansoni cercariae had the tails still attached when the bodies disappeared in the skin whereas all T. szidati cercariae shed their tails within 0-105 s after the onset of penetration movements. The faster invasion of T. szidati may result from the more sophisticated host-finding mechanisms of this species. Regarding S. mansoni, cercarial dermatitis, as immediate skin response, developed after a sensitization period of 19 days. PMID- 19543748 TI - Genetic diversity of Dientamoeba fragilis isolates of irritable bowel syndrome patients by high-resolution melting-curve (HRM) analysis. AB - Dientamoeba fragilis is a parasite that has been recognized as a causative agent of gastrointestinal symptoms. The search for genetic variation in D. fragilis based on the small-subunit (SSU) rRNA gene using restriction fragment length polymorphism was found not useful for molecular epidemiology. In this study, genetic variability of different clinical isolates of D. fragilis was explored by high-resolution melting curve (HRM) following polymerase chain reaction (PCR) in a one-step closed-tube method. Thirty fecal samples from irritable bowel syndrome (IBS) patients having D. fragilis trophozoites and negative for other organisms were involved in this study. According to the type of diarrhea, eight patients had acute, 14 patients had chronic intermittent, and eight patients had diarrhea alternating with constipation. HRM proved that four profiles (subtypes) were present as detecting by scanning mutation. One of these profiles (profile 1) was predominant (50%). Profile 2 was present on 20%. Profiles 3 and 4 were present on 16.7% and 13.4%, respectively. No mixed profiles were detected among the samples. The melting curves characterized by T(m)1=77.17+/-0.29 degrees C in profile 1, T(m)1=77.37+/-1.45 degrees C in profile 2, T(m)1=74.24+/-0.08 degrees C and T(m)2=79.64+/-0.09 degrees C in profile 3, and T(m)1=75.51 +/- 0.09 degrees C and T(m)=79.42 +/- 0.09 degrees C in profile 4. The relation between these profiles and types of diarrhea proved that the majority of patients having profile 1 (73.4%) and profile 4 (75%) had chronic intermittent diarrhea. All of the patients having profile 2 had acute diarrhea while all of the patients having profile 3 had diarrhea alternating with constipation. Although profile 1 was detected among all types of diarrhea, it was corresponding to 11/14 of patients with chronic intermittent diarrhea. All the differences were clinically and statistically significant. In conclusion, HRM following PCR was proved as a wide variation on D. fragilis genotypes that could be related to the characters of diarrhea among IBS patients. As the differences in HRM reflect different sequences of SSU RNA gene, thus, another study for identifying the sequences of these isolates (profiles) will be done and published later. PMID- 19543750 TI - Variation in moss-associated nitrogen fixation in boreal forest stands. AB - Traditionally it has been thought that most boreal forest communities lack a significant input of biologically fixed nitrogen. Recent discoveries of nitrogen fixation by cyanobacteria associated with mosses have resulted in a re-evaluation of this view. While it is recognized that rates of nitrogen fixation in mosses can be highly variable, there is little understanding as to why this occurs. I monitored nitrogen fixation, using acetylene reduction, in wet lowland and dry upland boreal forest communities, in central Canada, over a growing season. At the peak of nitrogen fixation in mid summer, Sphagnum capillifolium had an 11 times higher rate of fixation than Pleurozium schreberi. Variation in canopy openness and precipitation had no effect on rates of fixation over the growing season. In P. schreberi fixation rates did not vary between sites. Temperature had a positive effect on fixation rates in both S. capillifolium and P. schreberi, but the effect was 4 times more pronounced in S. capillifolium. Seasonal rates of nitrogen fixation were estimated at 193 mg N m(-2) for S. capillifolium and 23 mg N m(-2) for P. schreberi. With moderate increases in climate warming, predicted increases in nitrogen fixation in S. capillifolium are sufficient to raise its decomposition rate. Increased temperatures may therefore act synergistically to change boreal systems from a sink to a source of carbon. PMID- 19543749 TI - Quantification and organization of WIS2-1A and BARE-1 retrotransposons in different genomes of Triticum and Aegilops species. AB - A real-time PCR approach was adopted and optimized to estimate and compare, through a relative quantification, the copy number of WIS2-1A and BARE-1 retrotransposons. The aim of this approach was to identify and quantify the presence of these retrotransposons in Triticum and Aegilops species, and to understand better the genome organization of these retroelements. The species were selected to assess and compare the evolution of the different types of genomes between the more recent species such as the diploid Triticum monococcum, tetraploid T. dicoccon and hexaploid T. spelta, and the corresponding genome donors of the ancient diploids Aegilops (Ae. speltoides, Ae. tauschii, Ae. sharonensis and Ae. bicornis) and T. urartu. The results of this study indicated the presence of great variation in copy number both within and among species, and the existence of a non-linear relationship between retrotransposon copy number and ploidy level. For WIS2-1A, as expected, T. monococcum showed the lowest copy number which instead was similar in T. dicoccon and T. spelta; also T. urartu (AA), Ae. speltoides (BB) and Ae. tauschii (DD) showed a higher WIS2-1A copy number. Similar results were observed for BARE-1 retroelements except for Ae. tauschii which as in T. monococcum showed lower retroelements content; a similar content for T. dicoccon and T. urartu, whereas a higher number was found in T. spelta and Ae. speltoides. The results presented here are in accord with previous studies and contribute to unravelling the structure and evolution of polyploidy and repetitive genomes. PMID- 19543751 TI - Cybernetic modeling based on pathway analysis for Penicillium chrysogenum fed batch fermentation. AB - A macrokinetic model employing cybernetic methodology is proposed to describe mycelium growth and penicillin production. Based on the primordial and complete metabolic network of Penicillium chrysogenum found in the literature, the modeling procedure is guided by metabolic flux analysis and cybernetic modeling framework. The abstracted cybernetic model describes the transients of the consumption rates of the substrates, the assimilation rates of intermediates, the biomass growth rate, as well as the penicillin formation rate. Combined with the bioreactor model, these reaction rates are linked with the most important state variables, i.e., mycelium, substrate and product concentrations. Simplex method is used to estimate the sensitive parameters of the model. Finally, validation of the model is carried out with 20 batches of industrial-scale penicillin cultivation. PMID- 19543752 TI - Tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 in the cerebrospinal fluid of patients with cervical myelopathy and lumbar radiculopathy. AB - There have been few reports describing cytokines in the cerebrospinal fluid (CSF) of patients with spinal degenerative disorders. This study investigated whether interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) could be detected in CSF of patients with cervical myelopathy or lumbar radiculopathy and whether the concentrations of those cytokines correlated with the severity of disease conditions. CSF samples were obtained from 21 patients with cervical myelopathy (Group M) and 19 patients with lumbar radiculopathy (Group R), and six volunteers (control). The concentration of IL-6 was significantly higher in Groups M and R than in the control, possibly demonstrating spinal cord and nerve root damage, respectively. However, TNF-alpha was lower than the detection limit. IL-1beta was detected in only five samples from three patients in Group M and two volunteers in the control. The concentrations of IL-6 did not show any correlation with symptom duration, the scoring system by the Japanese Orthopaedic Association, or the duration of nerve root block. There is a possibility that the concentration of inflammatory cytokines in CSF can indicate certain pathological aspects of cervical myelopathy or lumbar radiculopathy. PMID- 19543753 TI - In vivo range of motion of the lumbar spinous processes. AB - The study design included an in vivo laboratory study. The objective of the study is to quantify the kinematics of the lumbar spinous processes in asymptomatic patients during un-restricted functional body movements with physiological weight bearing. Limited data has been reported on the motion patterns of the posterior spine elements. This information is necessary for the evaluation of traumatic injuries and degenerative changes in the posterior elements, as well as for improving the surgical treatment of spinal diseases using posterior procedures. Eight asymptomatic subjects with an age ranging from 50 to 60 years underwent MRI scans of their lumbar segments in a supine position and 3D models of L2-5 were constructed. Next, each subject was asked to stand and was positioned in the following sequence: standing, 45 degrees flexion, maximal extension, maximal left and right twisting, while two orthogonal fluoroscopic images were taken simultaneously at each of the positions. The MRI models were matched to the osseous outlines of the images from the two orthogonal views to quantify the position of the vertebrae in 3D at each position. The data revealed that interspinous process (ISP) distance decreased from L2 to L3 to L4 to L5 when measured in the supine position; with significantly higher values at L2-3 and L3 4 compared with L4-5. These differences were not seen with weight-bearing conditions. During the maximal extension, the ISP distance at the L2-3 motion segment was significantly reduced, but no significant changes were detected at L3 4 and L4-5. During flexion the ISP distances were not significantly different than those measured in the MRI position at all segments. Going from the left to right twist positions, the L4-5 segment had greater amounts of ISP rotation, while all segments had similar ranges of translation in the transverse plane. The interspinous process distances were dependent on body posture and vertebral level. PMID- 19543754 TI - Comparison of neuropathic pain and neuronal apoptosis following nerve root or spinal nerve compression. AB - Altered dorsal root ganglion (DRG) function is associated with neuropathic pain following spinal nerve injury. However, compression of the cauda equina and dorsal rhizotomy proximal to the DRG do not induce significant pain, whereas in the spinal nerve and peripheral nerve, injury distal to the DRG does induce neuropathic pain. Caspase signaling induces apoptosis, and caspase inhibitors prevent pain-related behavior. The degree of DRG neuronal apoptosis is thought to play a role in pain behavior. We suggest that differences in pain behavior according to the injury sites within the DRG may be related to imbalances in apoptotic injuries. The aim of this study was to determine which compression injury was more painful and to compare behavior with expression of tumor necrosis factor (TNF)-alpha in DRG and apoptosis in the DRG following crush injury to the L5 nerve root or L5 spinal nerve. Sprague-Dawley rats received a crush injury to the L5 spinal nerve (distal to the DRG), crush injury to the L5 nerve root (proximal to the DRG), or no crush injury (sham). Mechanical allodynia was determined by the von Frey test. Expression of TNF-alpha was compared among three groups using immunoblot findings. Furthermore, we compared the percentage of neurons injured in the DRG using immunostaining for apoptotic cells and localization of activated caspase 3. Mechanical allodynia was observed in both crush injury groups. The duration of mechanical allodynia in the distal crush group was significantly longer than in the proximal crush group (P < 0.05). TNF alpha expression was increased in DRG neurons following injury. DRG apoptosis in the distal crush group was significantly higher than in the proximal group at each time point (P < 0.05). This study suggests that spinal nerve crush injuries produce a greater degree of DRG apoptosis than do corresponding nerve root crush injuries, and that the former injuries are associated with longer lasting mechanical allodynia. Thus, differences in the time course of mechanical allodynia might be associated with an imbalance in DRG apoptosis. PMID- 19543755 TI - Docking and 3D QSAR studies of protoporphyrinogen oxidase inhibitor 3H pyrazolo[3,4-d][1,2,3]triazin-4-one derivatives. AB - Docking and three dimensional quantitative structure - activity relationship (3D QSAR) studies have been performed for protoporphyrinogen oxidase (PPO) inhibitor 3H-pyrazolo[3,4-d][1,2,3]triazin-4-one analogues which are potential herbicides to protect agricultural products from unwanted weeds. The 3D-QSAR studies have been carried out using shape, spatial, electronic and molecular field descriptors along with a few structural parameters. The chemometric tools used for the analyses are genetic function approximation (GFA), partial least squares (PLS) and genetic partial least squares (G/PLS). The whole data set (n = 34) was divided into a training set (75% of the data set) and a test set (remaining 25%) on the basis of K-means clustering technique applied on topological, spatial and electronic descriptor matrix. Models developed from the training set were used to predict the activity of the test set compounds. All the models have been validated internally, externally and by Y-randomization technique. Docking studies suggest that the molecules bind with a hydrophobic pocket of the enzyme formed by some nonpolar amino acid (Ile168, Ile311, Ile412, Met365, Phe65 and Val164) residues. The QSAR studies suggest that for better activity the molecules should have symmetrical shape in the 3D space. For better PPO inhibitory activity, there should be a balance between the electrophilic and nucleophilic characters of the inhibitors. The charged surface area descriptors suggest that, the positive charge distributed over a large surface area may enhance the activity. Molecular field probes reflect that increase in steric volume and positively charged surface area may enhance the herbicidal activity. PMID- 19543756 TI - What do dolphins (Tursiops truncatus) understand about hidden objects? AB - Object permanence, the ability to mentally represent and reason about objects that have disappeared from view, is a fundamental cognitive skill that has been extensively studied in human infants and terrestrial animals, but not in marine animals. A series of four experiments examined this ability in bottlenose dolphins (Tursiops truncatus). After being trained on a "find the object" game, dolphins were tested on visible and invisible displacement tasks, and transpositions. In Experiments 1 and 2, dolphins succeeded at visible displacements, but not at invisible displacements or transpositions. Experiment 3 showed that they were able to pass an invisible displacement task in which a person's hand rather than a container was used as the displacement device. However, follow-up controls suggested they did so by learning local rules rather than via a true representation of the movement of hidden objects. Experiment 4 demonstrated that the dolphins did not rely on such local rules to pass visible displacement tasks. Thus, like many terrestrial animals, dolphins are able to succeed on visible displacement tasks, but seem unable to succeed on tasks requiring the tracking of hidden objects. PMID- 19543757 TI - CT evaluation of subarachnoid hemorrhage: a practical review for the radiologist interpreting emergency room studies. AB - Radiologists working in an emergency radiology setting frequently interpret computed tomography (CT) studies of patients with suspected subarachnoid hemorrhage (SAH). This article reviews the sensitivity of CT for detection of SAH, some major patterns of SAH related to a ruptured aneurysm, and the differential diagnosis of SAH not due to aneurysmal rupture. PMID- 19543758 TI - Phytoremediation and phytosensing of chemical contaminants, RDX and TNT: identification of the required target genes. AB - High explosives such as hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), and 2,4,6 trinitrotoluene (TNT) are important contaminants in the environment and phytoremediation has been viewed as a cost-effective abatement. There remains, however, an insufficient knowledge-base about how plants respond to explosives, especially in the steady state. Microarray analysis was conducted on Arabidopsis thaliana that were grown in Murashige and Skoog media containing steady-state levels of 0.5 mM RDX or 2.0 microM TNT to study the effect of these compounds on its transcriptional profile. Our results for both RDX and TNT were consistent with the existing theory for xenobiotic metabolism in plants. Among the genes that were differentially expressed included oxidoreductases, cytochrome P450s, transferases, transporters, and several unknown expressed proteins. We discuss the potential role of upregulated genes in plant metabolism, phytoremediation, and phytosensing. Phytosensing, the detection of field contamination using plants, is an end goal of this project. PMID- 19543760 TI - ESTROM 2008 International Conference. PMID- 19543759 TI - Screening pentachlorophenol degradation ability by environmental fungal strains belonging to the phyla Ascomycota and Zygomycota. AB - Pentachlorophenol (PCP) bioremediation by the fungal strains amongst the cork colonising community has not yet been analysed. In this paper, the co- and direct metabolism of PCP by each of the 17 fungal species selected from this community were studied. Using hierarchical data analysis, the isolates were ranked by their PCP bioremediation potential. Fifteen isolates were able to degrade PCP under co metabolic conditions, and surprisingly Chrysonilia sitophila, Trichoderma longibrachiatum, Mucor plumbeus, Penicillium janczewskii and P. glandicola were able to directly metabolise PCP, leading to its complete depletion from media. PCP degradation intermediates are preliminarily discussed. Data emphasise the significance of these fungi to have an interesting potential to be used in PCP bioremediation processes. PMID- 19543761 TI - Skeletal tuberculosis in children in the Western world: 18 new cases with a review of the literature. AB - PURPOSE: The occurrence of pediatric skeletal (extra-spinal) tuberculosis in the developed world is extremely rare. The purpose of this study was to review the cases at our institutions. METHODS: We performed a retrospective review of all pediatric biopsy-proven skeletal (extra-spinal) cases of tuberculosis over a five year period. RESULTS: Eighteen patients of biopsy-proven tuberculosis were identified. The mean age was 12 years (range 7-20). Lesion locations included: distal humerus, ulna, scapula, acetabulum, proximal femur, proximal tibia, distal tibia, and calcaneus. All had symptoms of pain, swelling, and stiffness. Five cases had multi-focal involvement. Twenty-four lesions were noted in 18 patients. Nineteen lesions were cystic in nature at presentation. The sedimentation rate was normal in six and purified protein derivative (PPD) was negative in five patients. All received chemotherapy. Six patients underwent surgery. CONCLUSIONS: The diagnosis of pediatric skeletal tuberculosis can be made with good correlation of clinical, radiographic, and laboratory findings. Biopsy and culture are the gold standards in diagnosis. Prognosis is good with chemotherapy and non-operative management. Surgical intervention may be needed in select cases. PMID- 19543764 TI - Age- and gender-dependent changes in three-dimensional microstructure of cortical and trabecular bone at the human femoral neck. AB - We investigated age- and gender-related variation of both cortical and trabecular microstructure in human femoral neck. We found that age-related change of cortical porosity is more noticeable than that of trabecular parameter. Our data may help to gain more insight into the potential mechanism of osteoporotic femoral neck fractures. INTRODUCTION: Variations in the microstructure of cortical and trabecular bone contribute to decreased bone strength. Age- and gender-related changes in cortical and trabecular microstructure of femoral neck is unclear. The aim of this study was to identify three-dimensional (3D) microstructural changes of both cortical and trabecular bone simultaneously in human femoral neck with age and gender, using micro-computed tomography (micro CT). We hypothesized that there would be differences in age-related changes of cortical and trabecular bone for both women and men. METHODS: We used 56 femoral necks of 28 women and men (57-98 years of age) from a Japanese population. The subjects were chosen to give an even age and gender distribution. Both women and men were divided into three age groups: middle (57-68 years), old (72-82 years), and elderly (87-98 years) groups. We examined cortical bone specimen from the inferior sector of femoral neck and trabecular bone specimen from the middle of femoral neck using micro-CT and 3D bone analysis software. RESULTS: Cortical thickness (Ct.Th) decreased by 10-15%, cortical porosity (Ca.V/TV) almost doubled, and canal diameter (Ca.Dm) increased by 65-77% between the middle-aged and elderly groups for both women and men. The trabecular bone volume fraction (BV/TV) decreased by around 20%; trabecular thickness (Tb.Th), trabecular number (Tb.N), and connectivity density (Conn.D) decreased; and trabecular separation (Tb.Sp) and structure model index (SMI) increased with age for both women and men. As compared with women, men had higher Ct.Th and BV/TV and lower Ca.V/TV and Ca.Dm among three age groups. There was a significant inverse correlation between Ca.V.TV and BV/TV for both women and men. CONCLUSION: Our findings indicate that Ct.Th and BV/TV decreased, and Ca.V/TV and Ca.Dm increased in femoral neck with age for both women and men. The most obvious age-related change is the increase of Ca.V/TV. The decrease of BV/TV with age is more noticeable than that of Ct.Th. This is the first study that has provided both cortical and trabecular microstructural data simultaneously in a Japanese sample. These data may help us to gain more insight into the potential mechanism of osteoporotic femoral neck fractures. PMID- 19543767 TI - Mean platelet volume in Henoch-Schonlein purpura: relationship to gastrointestinal bleeding. AB - Henoch-Schonlein purpura (HSP) is the most common systemic vasculitis in children. Gastrointestinal (GI) bleeding is one of the major complications affecting one third of the cases which may cause serious morbidity. Platelet volume directly correlates with the platelet function and activation. Small platelets have lower functional capabilities than larger ones. The aim of this retrospective study was to evaluate levels of mean platelet volume (MPV) in patients with HSP compared with healthy controls and to investigate the relationship between MPV and gastrointestinal bleeding. The study consisted of 43 HSP patients (male/female = 25/18, mean age = 6.2 +/- 2.6 years) and 27 age matched healthy children (male/female = 14/13, mean age = 6.9 +/- 2 years) as control group. HSP patients had significantly lower MPV levels than healthy controls (7.5 +/- 0.8 vs. 7.9 +/- 0.5, p = 0.027). Thirteen of 43 patients had gastrointestinal bleeding. MPV was significantly lower in patients with GI bleeding than patients without bleeding (7.0 +/- 0.8 vs. 7.7 +/- 0.6, p = 0.01). Platelet counts, white blood cell counts, and C-reactive protein levels were significantly higher in patients with GI bleeding when compared to patients without GI bleeding (p = 0.03, p = 0.004, and p = 0.03, respectively). This study suggests that low MPV may contribute to GI bleeding in HSP. PMID- 19543766 TI - Association analyses of vitamin D-binding protein gene with compression strength index variation in Caucasian nuclear families. AB - This study was conducted to test whether there exists an association between vitamin D-binding protein (DBP) gene and compression strength index (CSI) phenotype. Candidate gene association analyses were conducted in total sample, male subgroup, and female subgroup, respectively. Two single-nucleotide polymorphisms (SNPs) with significant association results were found in males, suggesting the importance of DBP gene polymorphisms on the variation in CSI especially in Caucasian males. INTRODUCTION: CSI of the femoral neck (FN) is a newly developed phenotype integrating information about bone size, body size, and bone mineral density. It is considered to have the potential to improve the performance of risk assessment for hip fractures because it is based on a combination of phenotypic traits influencing hip fractures rather than a single trait. CSI is under moderate genetic determination (with a heritability of approximately 44% found in this study), but the relevant genetic study is still rather scarce. METHODS: Based on the known physiological role of DBP in bone biology and the relatively high heritability of CSI, we tested 12 SNPs of the DBP gene for association with CSI variation in 405 Caucasian nuclear families comprising 1,873 subjects from the Midwestern US. Association analyses were performed in the total sample, male and female subgroups, respectively. RESULTS: Significant associations with CSI were found with two SNPs (rs222029, P = 0.0019; rs222020, P = 0.0042) for the male subgroup. Haplotype-based association tests corroborated the single-SNP results. CONCLUSIONS: Our findings suggest that the DBP gene might be one of the genetic factors influencing CSI phenotype in Caucasians, especially in males. PMID- 19543768 TI - Surgical approach with Er:YAG laser on osteonecrosis of the jaws (ONJ) in patients under bisphosphonate therapy (BPT). AB - Osteonecrosis of the jaw (ONJ) in patients on long-term bisphosphonate Therapy (BPT) has been reported with increasing frequency in literature over the past 4 years. Therapy for this condition is still a dilemma. Temporary suspension of BPT offers no short-term benefits; hyperbaric oxygen has no proven efficacy and therefore is not recommended. Intermittent or continuous antibiotic therapy with surgical debridement can be beneficial to palliate the symptoms. Er:YAG laser can be used to eliminate necrotic bone portions by partial or total resection as an alternative to conventional rotary devices. In our study, 91 patients affected by ONJ-BP lesion, for a total of 115 ONJ sites were observed between January 2004 and May 2008 (Department of Odontostomatology, University of Parma). Fifty-five ONJ sites were considered for this study in four different groups, retrospectively identified on the basis of treatment performed (G1-G4). G1: 13 ONJ-BP sites were treated with medical therapy (amoxicillin 1gr x 3/die per os with metronidazole 250 mg x 2/die per os) for at least 2 weeks; G2: 17 ONJ-BP sites received medical treatment in association with cycles of low-level laser therapy (LLLT) applications performed using an Nd:YAG laser (1,064 nm) once a week for 2 months; G3: 13 ONJ-BP sites were surgically treated (sequestrectomy of necrotic bone, debridement, corticotomy/surgical removal of alveolar and/or cortical bone); G4: 12 ONJ-BP sites were treated with surgical therapy performed using an Er:YAG laser (2,940 nm) in association with LLLT. Clinical success has been defined for each treatment performed as: (a) complete mucosal healing free from signs and symptoms (classified as stage "0") or (b) transition from a higher to a lower stage (Ruggiero staging) for at least 3 months. All the ONJ-BP sites treated with Er:YAG laser (G4 group) had a clinical improvement (100%) and 87.5% of sites had a complete mucosal healing with a mean follow-up of 13 months. The result obtained in the G4 is extremely significant in comparison with those obtained by medical treatment alone or in a traditional surgical approach. Thanks to the high degree of affinity of this wavelength for water and hydroxyapatite, both soft and bone tissues can be easily treated. This technique can also be used for conservative operations whereby necrotic bone is vaporized until healthy bone is reached. In addition, an additional advantage of the Er:YAG laser is its bactericidal and possible biostimulatory action, accelerating the healing of both soft tissues and bone tissues, in comparison to conventional treatments. In conclusion, from our experience, it is possible to observe that an early conservative surgical approach with Er:YAG laser associated with LLLT, for BP induced ONJ could be considered as more efficient in comparison with medical therapy or other conventional techniques. PMID- 19543765 TI - Global vitamin D status and determinants of hypovitaminosis D. AB - This review describes the vitamin D status in different regions of the world with the objective of understanding the scope of hypovitaminosis D and the factors related to its prevalence that may contribute to the pathogenesis of osteoporosis and fragility fractures. INTRODUCTION: Vitamin D status has been linked to the pathogenesis of hip fractures as well as other skeletal and non-skeletal disorders. The purpose of this review is to provide a global perspective of vitamin D status across different regions of the world and to identify the common and significant determinants of hypovitaminosis D. METHODS: Six regions of the world were reviewed-Asia, Europe, Middle East and Africa, Latin America, North America, and Oceania-through a survey of published literature. RESULTS: The definition of vitamin D insufficiency and deficiency, as well as assay methodology for 25-hydroxyvitamin D or 25(OH)D, vary between studies. However, serum 25(OH)D levels below 75 nmol/L are prevalent in every region studied whilst levels below 25 nmol/L are most common in regions such as South Asia and the Middle East. Older age, female sex, higher latitude, winter season, darker skin pigmentation, less sunlight exposure, dietary habits, and absence of vitamin D fortification are the main factors that are significantly associated with lower 25(OH)D levels. CONCLUSION: Reports from across the world indicate that hypovitaminosis D is widespread and is re-emerging as a major health problem globally. PMID- 19543769 TI - Carotid artery dissection following a minor household accident in a 10-month-old child. AB - INTRODUCTION: Dissection of the carotid arteries is an important cause of ischaemic stroke in childhood. Rarely, blunt trauma to the soft palate can result in an internal carotid artery (ICA) dissection leading to thrombosis, embolism and stroke. CASE PRESENTATION: A 10-month-old child was admitted to hospital having presented with minor intra-oral trauma following a fall from a sofa whilst carrying a spoon in her mouth. After an interval, the child's level of consciousness decreased. Computed tomography and magnetic resonance imaging of the brain and neck revealed occlusion of the left ICA, oedema of the perivascular tissues and cerebral ischaemia in the left middle cerebral artery territory. CONCLUSION: A carotid artery dissection after minor trauma to the soft palate is a rare, but potentially severe complication. The combination of minor precipitating trauma, prolonged symptom-free intervals and complexity of clinical neurological assessment in small children contributes to a difficult diagnostic dilemma. PMID- 19543770 TI - Comparison of age-dependent quantitative changes in the male labial gland secretion of Bombus terrestris and Bombus lucorum. AB - Age-related changes of antennal-active components of male labial gland extracts were studied in two closely related bumblebee species, Bombus terrestris and B. lucorum. In B. terrestris, compounds eliciting electroantennogram (EAG) responses of virgin queens were ethyl dodecanoate, 2,3-dihydrofarnesal, 2,3 dihydrofarnesol, hexadecan-1-ol, octadeca-9,12,15-trien-1-ol, and geranylcitronellol. Compounds that elicited EAG responses from queens of B. lucorum were ethyl dodecanoate, ethyl tetradec-7-enoate, ethyl tetradec-9-enoate, ethyl hexadec-9-enoate, hexadecan-1-ol, hexadec-7-enal, octadeca-9,12-dien-1-ol, octadeca-9,12,15-trien-1-ol, and octadecan-1-ol. Quantities of these compounds in the labial glands changed significantly over the lifetime of the respective males of the two species. In both species, concentrations of the respective compounds reached their maximum within seven days after eclosion. Subsequently, a rapid decrease in the amount of EAG-active compounds occurred in B. terrestris, whereas in B. lucorum the amount of active compounds stayed approximately constant or decreased at a slow rate. Microscopy showed that in B. terrestris secretory cells of the labial glands undergo apoptosis from the fifth to the tenth day of life, whilst in B. lucorum labial gland cells remain unchanged throughout the life of the males. PMID- 19543771 TI - Predictive factors of regional toxicity and serum creatine phosphokinase levels after isolated limb infusion for melanoma: a multi-institutional analysis. AB - BACKGROUND: Isolated limb infusion (ILI) is a minimally invasive technique delivering regional chemotherapy to treat in-transit extremity melanoma. Determining perioperative factors that could predict toxicity is important to optimize strategies to improve clinical outcomes after regional chemotherapy in melanoma. METHODS: Perioperative factors from 171 ILI patients performed at eight centers from 2001 to 2008 were reviewed. The Wieberdink limb toxicity scale and creatine phosphokinase (CK) levels were used to measure toxicity. Logistic regression analysis was used to estimate the association between toxicity and perioperative parameters. RESULTS: Mild (grades I-II) and severe (grades >or=III) limb toxicity developed in 68% and 32% of patients, respectively. Melphalan adjusted for ideal body weight (aIBW) and papaverine were used in 47% and 63% of patients, respectively. Median peak CK for all patients was 563 U/l, and median peak occurred at postoperative day 4. On univariate analysis, papaverine and high CK levels (>563 U/l) were significantly associated with higher toxicity. On the contrary, aIBW was significantly associated with a lower risk of severe toxicity. Perfusate blood gas at 30 min [pH, PaO(2), and base excess (BE) ], limb temperature, and ischemia time were not predictive of limb toxicity. On multivariate analysis, severe toxicity was associated with female sex (P = 0.01), papaverine (P = 0.01), and high peak CK levels (P < 0.01). Independent predictors of high CK levels included younger age, unadjusted melphalan dose, and low PaO(2) at 30 min. CONCLUSIONS: ILI can be performed with an acceptable morbidity. Papaverine use, female gender, and high peak CK were associated with higher limb toxicity. CK levels can be diminished significantly with aIBW. PMID- 19543773 TI - Isolated tumor cells in sentinel lymph node and clinical implications for early breast cancer. PMID- 19543774 TI - Dual tracer 11C-choline and FDG-PET in the diagnosis of biochemical prostate cancer relapse after radical treatment. AB - PURPOSE: The purpose of this study was to evaluate a dual tracer 2-deoxy-2-[F 18]fluoro-D: -glucose (FDG) and (11)C-choline positron emission tomography (PET) protocol in the detection of biochemical prostate cancer relapse. PROCEDURES: Seventy-three patients (median Prostate Specific Antigen (PSA) Test value 2.7 ng/ml (1.1-5.4)) after radical treatment. PET scans were performed by means of a ECAT-Exact HR+ in the first 18 patients and in a PET/computed tomography Biograph II in the remaining 55 patients. RESULTS: The sensitivity of (11)C-choline and FDG was 60.6% and 31%. In PSA levels over 1.9 ng/ml, sensitivity increased to 80% and 40%, respectively. In the group receiving adjuvant hormone therapy, the diagnostic yields were 71.2% and 43%, respectively. While (11)C-choline-PET could not differentiate well and poorly differentiated Gleason score patients, FDG-PET results were almost significant (p = 0.058). CONCLUSIONS: A PSA value higher than 1.9 ng/ml determines a significant increase in the diagnostic yield. Adjuvant hormonotherapy has no influence on the PET results. FDG has a better correlation with the Gleason score than (11)C-choline. PMID- 19543775 TI - Intravital imaging of tumor apoptosis with FRET probes during tumor therapy. AB - PURPOSE: The aim of the study is to dynamically and non-invasively monitor the apoptosis events in vivo during photodynamic therapy (PDT) and chemotherapy. PROCEDURES: A FRET probe, SCAT3, was utilized to determine activation of caspase 3 during tumor cell apoptosis in mice, induced by PDT, and cisplatin treatments. Using this method, dynamics of caspase-3 activation was observed both in vitro and in vivo. RESULTS: Analysis of the fluorescent missions from tumor cells indicated that the caspase-3 activation started immediately after PDT treatment. In contrast, the caspase-3 activation started about 13 and 36 h after cisplatin treatment in vitro and in vivo, respectively. CONCLUSIONS: FRET could be used effectively to monitor activation of caspase-3 in living organism. This method could be used to provide rapid assessment of apoptosis induced by anti-tumor therapies for improvement of treatment efficacy. PMID- 19543777 TI - Favorable outcome of percutaneous repair of achilles tendon ruptures in the elderly. AB - Percutaneous repair of Achilles tendon (AT) ruptures reportedly reduces the risk of rerupture compared to nonoperative treatment and reduces the risk of wound infection compared to open surgery. We retrospectively reviewed the postoperative Achilles tendon total rupture score (ATRS), and the maximum calf circumference in 35 patients over 65 years of age who sustained an acute tear of the AT and underwent percutaneous repair of the AT. There were 26 men and nine women with a mean age at operation of 73.4 +/- 8.7 years (range, 65-86 years). Of the 35 recruited patients, we report on 27 patients for whom we have a full data set. The minimum followup was 49 months (mean, 88 months; range, 49-110 months). The ATRS had a postoperative average rating of 69.4 +/- 14 (range, 56-93). All patients were able to bear weight fully on the affected limb by the eighth postoperative week. The data suggest that percutaneous repair of the AT is a suitable option for patients older than 65, producing similar outcomes when compared to percutaneous repair in younger patients of previous reports. LEVEL OF EVIDENCE: Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19543779 TI - Infection rate after spine surgery in cerebral palsy is high and impairs results: multicenter analysis of risk factors and treatment. AB - Infection is a serious complication of surgery to correct scoliosis in patients with cerebral palsy (CP). We obtained multicenter representative figures for deep and superficial infection rates, analyzed risk factors and treatment outcomes, and compared deformity correction relative to infection. We retrospectively reviewed 157 patients who had posterior spinal fusion for CP at one of eight centers. Preoperative and intraoperative variables were subjected to multivariate analysis to determine factors predictive of infection. There were 16 wound infections (10%; nine deep, seven superficial). Only two study factors predicted infection: higher preoperative white blood cell count (8.5 versus 6.4 [in those without infection] x 10(3)) and use of a unit rod (15% versus 5% for bent rods). Fourteen patients underwent irrigation and debridement procedures. Five infections required 2 months or longer to resolve. Two had implant removal. Final curve correction was lower for those with deep infections than those without (67% versus 53%, respectively). We noted a trend toward greater percentages of pain at last followup in those with deep infection than in those without infection (50% versus 18%, respectively) but the study was not adequately powered to confirm this point. Our infection rate in scoliosis surgery for CP was higher than that for most elective spinal deformity surgery. LEVEL OF EVIDENCE: Level III, retrospective case-control study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19543778 TI - Improved tendon radioprotection by combined cross-linking and free radical scavenging. AB - Allograft safety is a great concern owing to the risk of disease transmission from nonsterile tissues. Radiation sterilization is not used routinely because of deleterious effects on the mechanical integrity and stability of allograft collagen. We previously reported several individual cross-linking or free radical scavenging treatments provided some radioprotective effects for tendons. We therefore asked whether a combination of treatments would provide an improved protective effect after radiation exposure regarding mechanical properties and enzyme resistance. To address this question we treated 90 rabbit Achilles tendons with a combination of cross-linking (1-ethyl-3-[3-dimethyl aminopropyl] carbodiimide [EDC]) and one of three scavenging regimens (mannitol, ascorbate, or riboflavin). Tendons then were exposed to one of three radiation conditions (gamma or electron beam irradiation at 50 kGy or unsterilized). Combination treated tendons (10 per group) had increases in mechanical properties and higher resistance to collagenase digestion compared with EDC-only and untreated tendons. Irradiated tendons treated with EDC-mannitol, -ascorbate, and -riboflavin combinations had comparable strength to native tendon and had averages of 26%, 39%, and 37% greater, respectively, than those treated with EDC-only. Optimization of a cross-linking protocol and free radical scavenging cocktail is ongoing with the goal of ensuring sterile allografts through irradiation while maintaining their structure and mechanical properties. PMID- 19543780 TI - Economic incentives to promote innovation in healthcare delivery. AB - Economics influences how medical care is delivered, organized, and progresses. Fee-for-service payment encourages delivery of services. Fee-for-individual service, however, offers no incentives for clinicians to efficiently organize the care their patients need. Global capitation provides such incentives; it works well in highly integrated practices but not for independent practitioners. The failures of utilization management in the 1990s demonstrated the need for a third alternative to better align incentives, such as bundling payment for an episode of care. Building on Medicare's approach to hospital payment, one can define expanded diagnosis-related groups that include all hospital, physician, and other costs during the stay and appropriate preadmission and postdischarge periods. Physicians and hospitals voluntarily forming a new entity (a care delivery team) would receive such bundled payments along with complete flexibility in allocating the funds. Modifications to gainsharing and antikickback rules, as well as reforms to malpractice liability laws, will facilitate the functioning of the care delivery teams. The implicit financial incentives encourage efficient care for the patient; the episode focus will facilitate measuring patient outcomes. Payment can be based on the resources used by those care delivery teams achieving superior outcomes, thereby fostering innovation improving outcomes and reducing waste. PMID- 19543781 TI - Incidence of postthrombotic syndrome in patients undergoing primary total knee arthroplasty for osteoarthritis. AB - Postthrombotic syndrome (PTS) is characterized by edema, venous ectasia, hyperpigmentation, varicose veins, venous ulceration, and pain with calf compression after deep venous thrombosis (DVT). We determined the incidence of PTS after DVT diagnosed on screening ultrasound in patients undergoing primary total knee arthroplasty (TKA) for osteoarthritis (OA). We retrospectively reviewed the records of 1406 patients who underwent primary TKA for osteoarthritis and compared the incidence of PTS in patients without and with DVT. All patients had postoperative screening ultrasound. From these 1406 patients we identified 66 (4.7%) who had DVT, 50 of whom had a minimum of 1 year followup (mean, 4.97 years; range, 1.00-7.53 years). PTS was diagnosed if any two of six signs were documented in the medical record. Three of 50 patients with DVT (6%) had signs consistent with PTS; two of these three had a DVT proximal to the soleal arch. Seven (8%) of 88 patients randomly chosen for primary TKA because of OA with similar mean age and gender, but without DVT, had signs of PTS. PTS does not seem to be a major sequela of DVT in patients undergoing primary TKA for OA. LEVEL OF EVIDENCE: Level III, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19543782 TI - The 2009 Frank Stinchfield Award: "Hip squeaking": a biomechanical study of ceramic-on-ceramic bearing surfaces. AB - We designed and implemented an in vitro bench test to simulate and identify potential biomechanical causes for hip squeaking with alumina ceramic-on-ceramic bearing surfaces. All bearings were third-generation alumina ceramic with a 32-mm head coupled with a 56-mm acetabular component with a 32-mm ceramic insert. Conditions for testing were normal gait, high load, stripe wear, stripe wear in extreme load, metal transfer, edge wear with extreme load, and microfracture. Each condition was tested two times in dry conditions and two times in a lubricated condition with 25% bovine serum. Squeaking was reproduced in all dry conditions. It occurred quickly with high load, stripe wear, or metal transfer. Once squeaking occurred, it did not stop. Squeaking disappeared for all conditions when a small amount of lubricant was introduced. In lubricated conditions, squeaking was only reproduced for the material transfer condition. Our observations suggest squeaking is a problem of ceramic-ceramic lubrication and that this noise occurs when the film fluid between two surfaces is disrupted. Material (metal) transfer was the only condition that led to squeaking in a lubricated situation. PMID- 19543785 TI - Reduction of neointimal hyperplasia in porcine coronary arteries by 2-deoxy-D glucose. AB - BACKGROUND: The drug eluting stents have been shown to play a substantial role in preventing in-stent restenosis. This study was initiated to determine the efficacy of 2-deoxy-D-glucose (2-DG) in an in-stent restenosis model for reducing neointimal hyperplasia after coronary stent placement. METHODS: In a porcine overstretch model, three kinds of stents were investigated (n = 12 per group): bare metal stents (BMS), rapamycin-eluted stents (RES), and BMS after intracoronary short-term application of 2-DG (DGS). After 42 days histomorphometric and histopathological analyses were performed. RESULTS: Neointimal thickness (BMS: 0.38 +/- 0.08, RES: 0.24 +/- 0.11, DGS: 0.15 +/- 0.01), area stenosis (BMS: 47.39 +/- 2.76, RES: 32.2 +/- 2.08, DGS: 29.30 +/- 2.98) did not differ after 42 days between the RES and DGS but were significantly lower as compared to BMS only. Lumen area (BMS: 3.15 +/- 1.53, RES: 4.37 +/- 1.72, DGS: 4.77 +/- 2.14) was significantly higher in the DGS group in comparison to the BMS group. The calculated injury and inflammation scores were similar and re-endothelialization was confirmed in all groups. CONCLUSIONS: This study could demonstrate that in porcine stent model neointimal hyperplasia and re endothelialization after application of 2-DG are comparable to those seen in RES. Thus, 2-DG might be a promising clinical application for coronary stent coating. PMID- 19543786 TI - ADIPOQ gene polymorphism rs1501299 interacts with fibre intake to affect adiponectin concentration in children: the GENe-Diet Attica Investigation on childhood obesity. AB - BACKGROUND: Adiponectin, an adipose-derived hormone with central and peripheral actions, is involved in the regulation of energy homeostasis. Interactions between genetic and environmental factors have been associated with decrease in circulating adiponectin leading to obesity. AIM: We investigated whether variants of the ADIPOQ gene encoding adiponectin interact with diet to predict serum adiponectin concentration. METHODS: A cross-sectional study of healthy school aged children of Greek origin (n = 991), aged 11.2 +/- 0.6 years was conducted in 2005-2006. DNA was genotyped for two SNPs [rs1501299 (n = 741) and rs17300539 (n = 713)] located in the ADIPOQ gene. Detailed dietary, behavioural, lifestyle, anthropometric and biochemical data were recorded for all participants. RESULTS: Both SNPs were in HWE. The rs1501299 (GG vs GT + TT) x fibre interaction was significantly associated with adiponectin concentration (P = 0.028). When fibre intake was low, GG homozygotes exhibited significantly higher adiponectin concentrations compared to T allele carriers (mean +/- SD = 5.1 +/- 2.7 vs 4.2 +/ 2.3; P = 0.020). CONCLUSIONS: In the present study, the rs1501299 x fibre interaction was significantly associated with adiponectin levels; in specific, GG homozygotes exhibited higher adiponectin levels compared to T carriers under conditions of lower fibre intake. PMID- 19543787 TI - Ischemic postconditioning: experimental models and protocol algorithms. AB - Ischemic postconditioning, a simple mechanical maneuver at the onset of reperfusion, reduces infarct size after ischemia/reperfusion. After its first description in 2003 by Zhao et al. numerous experimental studies have investigated this protective phenomenon. Whereas the underlying mechanisms and signal transduction are not yet understood in detail, infarct size reduction by ischemic postconditioning was confirmed in all species tested so far, including man. We have now reviewed the literature with focus on experimental models and protocols to better understand the determinants of protection by ischemic postconditioning or lack of it. Only studies with infarct size as unequivocal endpoint were considered. In all species and models, the duration of index ischemia and the protective protocol algorithm impact on the outcome of ischemic postconditioning, and gender, age, and myocardial temperature contribute. PMID- 19543788 TI - [Antithrombotic therapy in atrial fibrillation: when vitamin K antagonists? When aspirin? When heparin? When combinations of anticoagulant and antiplatelet drugs?]. AB - Morbidity and mortality associated with atrial fibrillation are mainly related to thromboembolic complications, particularly ischemic strokes. The prevention of thromboembolism is an important component in the management of patients with atrial fibrillation. The choice of optimum antithrombotic therapy for a given patient depends on the risk of thromboembolism, on the one hand, and the risk of intracerebral hemorrhage, on the other hand. Concerning the benefit-to-risk stratification the problem lies in the similar and sometimes even identical risk factors for both thromboembolism and haemorrhage.At present, oral vitamin K antagonists are recommended for patients with atrial fibrillation at moderate or high risk of ischemic stroke. The thromboembolic risk should be assessed using validated stratification schemes, such as the CHADS(2) score. Aspirin alone is recommended for patients at low risk of thromboembolic complications. A combination of anticoagulant and antiplatelet drugs is necessary in patients with atrial fibrillation undergoing percutaneous coronary intervention and stent implantation, but the optimal therapeutic management of these patients has not yet been defined. Hopefully, the development of new antithrombotic agents being easier to use and having a superior benefit-to-risk ratio will extend effective prevention of thromboembolic events to a greater part of the atrial fibrillation population at risk. PMID- 19543790 TI - Does ADHD moderate the manifestation of anxiety disorders in children? AB - The main aim of this study was to examine the moderating effects of attention deficit hyperactivity disorder (ADHD) on anxiety disorders in children. Data were analyzed from a large referred sample of children with anxiety disorder without comorbid ADHD (anxiety disorder, N = 253), anxiety disorder plus comorbid ADHD (anxiety disorder + ADHD, N = 704), and ADHD without comorbid anxiety disorder (ADHD, N = 511). Children were comprehensively assessed, including by structured diagnostic interview (K-SADS-E). Overall rates of individual anxiety disorders, as well as age of onset and severity of illness were not significantly different in the presence of comorbid ADHD. School functioning in children with anxiety disorders was negatively impacted by the presence of comorbid ADHD. Frequency of mental health treatment in children with anxiety disorders was significantly increased in the presence of comorbid ADHD. ADHD had a limited impact on the manifestation of anxiety disorder in children suggesting that ADHD and anxiety disorders are independently expressed. PMID- 19543789 TI - Alzheimer's disease, but not ageing or depression, affects dual-tasking. AB - Two experiments are reported that assess dual task performance in Alzheimer's disease (AD), in chronic depression and in healthy old age. Results suggest that dual task impairments are present in AD but are not shown in depression. This is true even when episodic memory performance is equated between the groups. These results, together with those of previous studies, point to dual task performance as an aid to diagnosis of AD relative to depression. This is of particular relevance when episodic memory tests cannot distinguish between the two conditions. The dual task paradigm appears to have considerable promise in assisting the early detection of the specific cognitive deficits associated with AD, and in monitoring their progression, both in the laboratory setting and in everyday tasks. Results also are of theoretical interest in pointing to a specific dual task coordination function in the healthy human cognitive system that allows for the coordination of two tasks performed simultaneously and which is damaged in AD but not in depression. PMID- 19543791 TI - ADHD in international adoptees: a national cohort study. AB - Several investigators have reported an increased frequency of attention/hyperactivity symptoms in international adoptees, though population based studies are lacking. In this national cohort study, we aimed to determine the prevalence of ADHD medication in international adoptees in Sweden, in comparison to the general population. A further purpose was to study gender, age at adoption and region of origin as predictors of ADHD medication in international adoptees. The study population consisted of all Swedish residents born in 1985-2000 with Swedish-born parents, divided into 16,134 adoptees, and a comparison population of 1,326,090. ADHD medications were identified in the Swedish Prescribed Drug Register during 2006. Logistic regression was used to calculate the odds ratios. The rates of ADHD medication were higher in international adoptees than in the comparison population for both boys (5.3 vs. 1.5% for 10-15-year olds) and girls (2.1 vs. 0.3% for 10-15-year olds). International adoptees from all regions of birth more often consumed ADHD medication compared with the majority population, but the age and sex adjusted odds ratios were particularly high for adoptees from Eastern Europe, Middle East/Africa and Latin America. Adjusting for maternal education and single parenthood increased the odds ratios even further. The risk also increased with higher age at adoption. Adoptees from Eastern Europe have a very high risk for ADHD medication. A structured identification and support programme should be tailored for this group. Adoptees from other regions have a more moderately increased risk, which should be communicated to adoptive parents and to professionals who care for adoptees in their clinical practice. PMID- 19543793 TI - A case of a fibroadenoma coexisting with an invasive lobular carcinoma in the breast. AB - A case of a fibroadenoma coexisting with an invasive lobular carcinoma of the breast in a 60-year-old female is presented, and its pathological features are correlated with high-resolution magnetic resonance imaging (HR-MRI) and other imaging findings. The patient presented with the chief complaint of having a palpable mass in her right breast for 3 months. Mammography revealed a lobular mass with a micro-lobulated margin, which suggested a malignant nature; however, it included coarse calcifications. Sonographic imaging and HR-MRI findings were compatible with malignant tumor. Cytology was performed, and the results indicated an invasive carcinoma. Breast-conserving surgery was performed as a curative operation. The pathological features revealed a fibroadenoma coexisting with an invasive lobular carcinoma. This case suggests that radiologists should always pay attention to the associated malignant imaging characteristics, such as the shape and border of the mass, whenever a mass demonstrates benign-like calcifications. PMID- 19543792 TI - Psychiatric and cognitive phenotype in children and adolescents with myotonic dystrophy. AB - Myotonic dystrophy type 1 (DM1) is the most frequent inherited neuromuscular disorder. The juvenile form has been associated with cognitive and psychiatric dysfunction, but the phenotype remains unclear. We reviewed the literature to examine the psychiatric phenotype of juvenile DM1 and performed an admixture analysis of the IQ distribution of our own patients, as we hypothesised a bimodal distribution. Two-thirds of the patients had at least one DSM-IV diagnosis, mainly attention deficit/hyperactivity disorder and anxiety disorder. Two-thirds had learning disabilities comorbid with mental retardation on one hand, but also attention deficit, low cognitive speed and visual spatial impairment on the other. IQ showed a bi-modal distribution and was associated with parental transmission. The psychiatric phenotype in juvenile DM1 is complex. We distinguished two different phenotypic subtypes: one group characterised by mental retardation, severe developmental delay and maternal transmission; and another group characterised by borderline full scale IQ, subnormal development and paternal transmission. PMID- 19543794 TI - Effect of cadmium on the distribution of hydroxyl radical, superoxide and hydrogen peroxide in barley root tip. AB - In the present study, we investigated the alteration of reactive oxygen species production along the longitudinal axis of barley root tips during Cd treatment. In unstressed barley root tips, H(2)O(2) production decreased from the root apex towards the differentiation zone where again, a slight increase was observed towards the more mature region of root. An opposite pattern was observed for O(2)(*-) and OH(*) generation. The amount of both O(2)(*-) and OH(*) was highest in the elongation zone, decreased in the root apex and at the differentiation zone of root, then increased again towards the more mature region of root. An elevated Cd-induced O(2)(*-) production started in the elongation zone and increased further along the differentiation zone of barley root tip. In contrast, Cd-induced H(2)O(2) production was localised to the root elongation zone and to the beginning of the differentiation zone. In contrast to Cd-induced H(2)O(2) and O(2)(*-) production, Cd reduced OH(*) production along the whole barley root tip. Our results suggest that not only an increase but also the spatial distribution of reactive oxygen species production is involved in the Cd-induced stress response of barley root tip. PMID- 19543795 TI - beta-Alanine elevates dopamine levels in the rat nucleus accumbens: antagonism by strychnine. AB - Glycine receptors (GlyRs) in the nucleus accumbens (nAc) have recently been suggested to be involved in the reinforcing and dopamine-elevating properties of ethanol via a neuronal circuitry involving the VTA. Apart from ethanol, both glycine and taurine have the ability to modulate dopamine output via GlyRs in the same brain region. In the present study, we wanted to explore whether yet another endogenous ligand for the GlyR, beta-alanine, had similar effects. To this end, we monitored dopamine in the nAc by means of in vivo microdialysis and found that local perfusion of beta-alanine increased dopamine output. In line with previous observations investigating ethanol, glycine and taurine, the competitive GlyR antagonist strychnine completely blocked the dopamine elevation. The present results suggest that beta-alanine has the ability to modulate dopamine levels in the nAc via strychnine-sensitive GlyRs, and are consistent with previous studies suggesting the importance of this receptor for modulating dopamine output. PMID- 19543796 TI - Validity of self-reported occupational noise exposure. AB - In all epidemiological studies the validity of self-reported questionnaire data is an important issue as the exposure assessment based on such data is a major source of bias in the risk estimation. A validation study was conducted based on a case-control study including 94 acoustic neuroma cases and 191 matched controls from the German Interphone Study to investigate the level of agreement between self-reported occupational noise exposure and a job-exposure-matrix (JEM) on noise exposure derived from a lifetime occupation calendar. The JEM was generated based on measurement data collected in the literature for various occupations. Level of agreement was investigated by using sensitivity, specificity, kappa coefficient and the Youden-Index. The receiver operating characteristics curve yielded an optimal cut point of 80 decibel(Acoustic) (dB(A)) to dichotomize noise exposure, displaying a moderate agreement between self-reported exposure and the JEM-based exposure (kappa of 0.53) that was slightly higher for cases than controls (kappas of 0.62 and 0.48). The agreement was only slightly lower if the longest held job or the last held job were used instead of the loudest job of the lifetime job history. The cut point of 80 dB(A) corresponds with regulations for workers safety with a recommendation to wear noise protection. The good levels of agreement between self-reported high occupational noise exposure compared with JEM-data, together with no substantial differences between cases and controls, suggest that self-reported data on occupational noise exposure is a valid exposure metric. Noise exposure appears to be appropriate if only exposure information on the last or the longest held job is available. PMID- 19543797 TI - Different fibre populations distinguished by their calcium transient characteristics in enzymatically dissociated murine flexor digitorum brevis and soleus muscles. AB - Enzymatically dissociated flexor digitorum brevis (FDB) and soleus fibres from mouse were used to compare the kinetics of electrically elicited Ca2+ transients of slow and fast skeletal muscle fibres, using the fast Ca2+ dye MagFluo4-AM, at 20-22 degrees C. For FDB two Ca2+ transient morphologies, types I (MT-I, 11 fibres, 19%) and II (MT-II, 47 fibres, 81%), were found, the kinetic parameters (amplitude, rise time, half width, decay time, and time constants of decay) being statistically different. For soleus (n = 20) only MT-I was found, with characteristics similar to MT-I from FDB. Correlations with histochemically determined mATPase, reduced nicotinamide adenine dinucleotide diaphorase and alpha-glycerophosphate dehydrogenase activities, as well as immunostaining and myosin heavy chain electrophoretic analysis of both muscles suggest that signals classified as MT-I may correspond to slow type I and fast IIA fibres while those classified as MT-II may correspond to fast IIX/D fibres. The results point to the importance of Ca2+ signaling for characterization of muscle fibres, but also to its possible role in determining fibre function. PMID- 19543798 TI - Reproducibility of cardiac output measurement by the nitrous oxide rebreathing technique. AB - Techniques for the measurement of cardiac output from soluble gas uptake by the lungs include the rebreathing method using nitrous oxide. The accuracy of this technique is well accepted, but its repeatability of measurement (precision) has not been well documented. We assessed the repeatability of measurements of pulmonary blood flow by the Innocor, a device employing the nitrous oxide rebreathing method. Successive paired measurements of pulmonary blood flow were made separated by a 5 min interval by the nitrous oxide rebreathing method, in 8 patients pre- or post cardiac surgery, and in 8 healthy volunteers. The standard deviation of the difference between first and second measurements was 0.84 l/min in the cardiac surgery group, and 1.25 l/min in the healthy volunteers. There was no significant bias in successive paired measurements of pulmonary blood flow in either the cardiac surgery patients (mean [95%CI] = -0.02 l/min [-0.62 to 0.57] or the healthy volunteers (0.00 l/min [-0.88 to 0.88]). Intra-class correlation coefficients for the healthy and cardiac patients were 0.77 and 0.64 respectively. Multiple measurements should be made and averaged when using the inert gas rebreathing technique for pulmonary blood flow determination. When comparing agreement with other methods for cardiac output measurement, the internal consistency of both methods should be considered. PMID- 19543799 TI - Trends in colorectal cancer incidence rates by age, race/ethnicity, and indices of access to medical care, 1995-2004 (United States). AB - OBJECTIVE: Colorectal cancer (CRC) incidence rates in the US decreased rapidly since 1998. This is largely thought to reflect increases in utilization of CRC screening through detection and removal of adenomatous polyps. However, the extent to which the decrease varies by age, race/ethnicity, and differences in access to medical care is largely unknown. METHODS: Temporal trends in CRC incidence rates were examined from 1995 to 2004 by regression analysis according to age (50-64, >= 65), race/ethnicity (whites, African Americans, and Hispanics), and categories of county-level indicators of access to care (poverty, primary care physician supply [PCP], uninsured rate [age 50-64], and metro/nonmetro) using incidence data from 19 cancer registries, covering about 53% of the US population. Changes in colorectal endoscopic screening and fecal occult blood stool test (FOBT) from 1995-1997 to 2002-2004 for the same set of county-level indicators were also analyzed, using data from the Behavioral Risk Factor Surveillance System (BRFSS). RESULTS: Among whites, CRC incidence rates decreased significantly from 1998 through 2004 in age >= 65, but not in age 50-64 in counties with high uninsured or poverty rates, fewer PCPs, or in nonmetro areas. Among African Americans or Hispanics, rates did not decrease in age 50-64 in general and age >= 65 in counties with high poverty rates, low PCP supply, and nonmetro counties (African Americans). Colorectal endoscopic screening rates increased significantly among whites in both age groups, but not among Hispanics (aged 50-64 in general and aged >= 65 residing in high poverty counties) or African Americans residing in counties with higher uninsured rates (age 50-64), low PCP supply, high poverty rates, and nonmetro counties (age >= 65). FOBT rates remained unchanged during the study time period. CONCLUSIONS: Our results suggest that individuals residing in poorer communities with lower access to medical care have not experienced the reduction in CRC incidence rates that have benefited more affluent communities; these disparities may be related to health care access barriers to colorectal endoscopic screening. PMID- 19543800 TI - A convenient one-pot synthesis of 3-amino-2, 5-dihydropyridazine and pyrimidine derivatives in the presence of high surface area MgO as a highly effective heterogeneous base catalyst. AB - A new and efficient method for the preparation of 3-amino-2,5-dihydropyridazines from three-component reactions of (phenyl-hydrazono)-propan-2-one, aldehydes, and malononitrile or ethyl cyanoacetate in the presence of magnesium oxide (MgO) as a highly effective heterogeneous base catalyst is presented. Also, the three component reactions of aldehydes, amidine systems, and malononitrile or ethyl cyanoacetate for the formation of 4-amino-5-pyrimidine carbonitrile and pyrimidinone derivatives, respectively, are investigated. The salient features of these methods include high conversions, short reaction times, cleaner reaction profiles, and the use of inexpensive and readily available catalyst. PMID- 19543801 TI - Comparison between volatile emissions from transgenic apples and from two representative classically bred apple cultivars. AB - While most risk assessments contrast a transgenic resistant to its isogenic line, an additional comparison between the transgenic line and a classically bred cultivar with the same resistance gene would be highly desirable. Our approach was to compare headspace volatiles of transgenic scab resistant apple plants with two representative cultivars (the isogenic 'Gala' and the scab resistance gene containing 'Florina'). As modifications in volatile profiles have been shown to alter plant relationships with non-target insects, we analysed headspace volatiles from apple plants subjected to different infection types by gas chromatography-mass spectrometry. Marked differences were found between healthy and leafminer (Phyllonorycter blancardella) infested genotypes, where emissions between the transgenic scab resistant line and the two cultivars differed quantitatively in four terpenes and an aromatic compound. However, these modified odour emissions were in the range of variability of the emissions recorded for the two standard cultivars that proved to be crucial references. PMID- 19543802 TI - Socio-economic factors influencing the use of acaricides on livestock: a case study of the pastoralist communities of Nakasongola District, Central Uganda. AB - A study was conducted in Nakasongola district to determine socioeconomic factors that influence the use of acaricides on livestock. The information was got through focus group discussions (FGDs) and use of a questionnaire. Questionnaire was administered to one hundred households. Acaricides were used to kill ticks and biting flies which transmit diseases and cause discomfort to livestock. But to a less extent was also done for cosmetic purposes. Most of the farmers were aware of the correct acaricide dilutions as recommended by the manufacturers but they ignored them. But through trial and error came up with their own dilutions, which they said were very cost-effective. Further, they experimented on concoctions of different acaricide mixes and came up with acaricide combinations which were more effective in killing ticks and flies. Veterinarians and acaricide manufacturing companies had called this a malpractice. On the contrary, this should be treated as an innovation by farmers in their endeavour to find a cheaper sustainable method of controlling ticks and flies. Further research should therefore be done on these working "malpractices". PMID- 19543803 TI - Sero-prevalence of bovine brucellosis and its risk factors in Jimma zone of Oromia Region, South-western Ethiopia. AB - A cross sectional sero-prevalence study was conducted on 1,595 cattle in Jimma zone, Ethiopia to investigate the status of bovine brucellosis and identify potential risk factors. Sera samples were analyzed using Rose Bengal Plate Test (RBPT) and Complement Fixation Test (CFT). The overall individual and herd level sero-prevalences were 3.1% (n = 1,595) and 15.0% (n = 227), respectively. The sero-prevalence of bovine brucellosis at individual animal level was significantly higher in non-pregnant (11.18%) than pregnant (2.77%) and lactating (22.35%) than non-lactating animals (2.46%). Moreover, significantly higher sero prevalence was observed in herds of larger sizes. Individual animal sero prevalence was also positively associated with the occurrence of abortion (26.98 and 1.54% in those with and without previous history of abortion, respectively). Generally, the sero-prevalence of bovine brucellosis found in Jimma area was not high and the sero-prevalence was closely associated with some of the risk factors considered at individual animal and herd level. PMID- 19543804 TI - Oxygen detection in biological systems. AB - This article presents a brief description of analytical tools for monitoring evolution and consumption of molecular dioxygen in biological organisms. Based on its nature as a gas and its physical and chemical properties of the ground state 3Sigma(g)O2; different approaches have been developed for quantitative determinations: (i) manometry, (ii) formation of titratable sediments, (iii) solid state electrodes, (iv) EPR oximetry, (v) luminescence quenching, (vi) biological sensoring, (vii) mass spectrometry and (viii) amperometry. Among these methods mass spectrometry and amperometry are of special relevance for studies on the mechanisms of photosynthetic dioxygen evolution. Mass spectrometry is described in the article of Beckman et al. in this special issue. Therefore, the major part of this contribution focuses on amperometric methods that are currently widely used. Two different types of electrodes are described: (i) Clark type electrode and (ii) Joliot-type electrode. The complementary advantages of both systems are outlined. A more detailed description comprises the potential of the Joliot-type electrode for mechanistic studies on the reactivity of the different redox states of the water oxidizing complex (WOC). PMID- 19543805 TI - Aflatoxin and ochratoxin production by Aspergillus species under ex vivo conditions. AB - Aspergillus species are increasingly important human pathogens. It is not known whether toxic metabolites of many of these pathogenic species can act as virulence factors in aspergillosis. We examined isolates of aflatoxin and ochratoxin-producing species for toxin production in ex vivo conditions. Seven of the 21 aflatoxin-producing isolates screened produced aflatoxin at 35 and 37 degrees C on the general medium yeast extract sucrose agar (YES). However, none of them produced toxin at these temperatures on brain heart infusion agar (BHA), a medium that mimics human tissue, or on BHA with modified pH or sugar levels. Six of the 12 ochratoxin-producing isolates examined produced toxin at 35 degrees C on YES. All three isolates of A. alliaceus produced ochratoxin on BHA or modified BHA at 37 degrees C. One strain of A. pseudoelegans produced a minute amount of ochratoxin on modified BHA at 37 degrees C. These data indicate that aflatoxin is an unlikely virulence, factor but that ochratoxin may be a potential virulence factor in aspergillosis. PMID- 19543806 TI - An organizational perspective on ethics as a form of regulation. AB - In this paper we propose a theoretical framework for analysing the history and function of ethics as a form of regulation. Ethics in the form of codes, rules and declarations, constitutes regulatory policies, and we wish to suggest analysing such policies from an organizational perspective. In many instances ethics policies are reactions to particular events involving harm of patients or research participants. As such they seem to come forward as solutions to specific problems. However, not all such events that instigate the making of new policies, and policies often have other effects and are used for other purposes than what we might expect from the events preceding them: when ethics takes on the form of policy making, the relationship between problems and solutions is more complex. We suggest that an organizational perspective on ethics codes, rules and declarations can deliver a relevant framework for future studies of the implications of wanting to address ethical problems through policy making. PMID- 19543807 TI - Development of a battery of instruments for detailed measurement of health status in patients with COPD in routine care: the Nijmegen Clinical Screening Instrument. AB - PURPOSE: To compose a battery of instruments that provides a detailed assessment of health status (HS) in COPD but that is applicable and clinically meaningful in routine care. METHODS: In a previous study, we developed the Nijmegen Integral Assessment Framework (NIAF) that organizes existing tests and instruments by the sub-domains of HS they measure. Based on clinical and statistical criteria (correlation coefficients and Cronbach alpha's) we selected for each sub-domain instruments from the NIAF. A COPD-study group was used to determine c-scores, and two control groups were used to determine the score ranges indicating normal functioning versus clinically relevant problems for each sub-domain. Existing questionnaire completion software (TestOrganiser) was adapted to enhance clinical applicability. RESULTS: The NCSI measures eleven sub-domains of physiological functioning, symptoms, functional impairment, and quality of life. The TestOrganiser automatically processes the data and produces the graphical PatientProfileChart, which helps to easily interpret results. This envisages the problem areas and discrepancies between the different sub-domains. CONCLUSION: The NCSI provides a valid and detailed picture of a patient's HS within 15-25 min. In combination with the PatientProfileChart, the NCSI can be used perfectly in routine care as screening instrument and as a guide in patient-tailored treatment. PMID- 19543808 TI - Predicting EQ-5D utility scores from the 25-item National Eye Institute Vision Function Questionnaire (NEI-VFQ 25) in patients with age-related macular degeneration. AB - PURPOSE: In this study, we explored different statistical approaches to identify the best algorithm to predict EQ-5D utility scores from the NEI-VFQ 25 in patients with age-related macular degeneration (AMD). METHODS: Ordinary least squares (OLS), Tobit, and censored least absolute deviation (CLAD) approaches were compared using cross-sectional data (primary dataset, n = 151) at screening from a phase I/II clinical trial in patients with AMD. Three models were specified in this study: full (includes all 12 dimensions of the NEI-VFQ 25), short (includes only the general health dimension and the composite score), and reduced model (using stepwise regression). To evaluate the predictive accuracy of the models, the mean absolute prediction error (MAPE), mean error, and root means squared error were calculated using in-sample cross-validation (within the primary dataset) and out-of-sample validation using an independent dataset (n = 393). The model that provided the lowest prediction errors was chosen as the best model. RESULTS: In-sample cross-validation and out-of-sample validation consistently demonstrated that, compared to other approaches, heteroscedasticity adjusted OLS produced the lowest MAPE (mean values were 0.1400, 0.1593, respectively) for the full model, while CLAD performed best for the short and reduced models (mean values were 0.1299, 0.1483, respectively). The normality and homoscedasticity assumptions of both OLS and Tobit were rejected. CLAD, however, can accommodate these particular violations. CONCLUSIONS: The CLAD-short model is recommended for producing the EQ-5D utility scores when only the NEI-VFQ 25 data are available. PMID- 19543810 TI - The three-dimensional structure of diaminopimelate decarboxylase from Mycobacterium tuberculosis reveals a tetrameric enzyme organisation. AB - The three-dimensional structure of the enzyme diaminopimelate decarboxylase from Mycobacterium tuberculosis has been determined in a new crystal form and refined to a resolution of 2.33 A. The monoclinic crystals contain one tetramer exhibiting D(2)-symmetry in the asymmetric unit. The tetramer exhibits a donut like structure with a hollow interior. All four active sites are accessible only from the interior of the tetrameric assembly. Small-angle X-ray scattering indicates that in solution the predominant oligomeric species of the protein is a dimer, but also that higher oligomers exist at higher protein concentrations. The observed scattering data are best explained by assuming a dimer-tetramer equilibrium with about 7% tetramers present in solution. Consequently, at the elevated protein concentrations in the crowded environment inside the cell the observed tetramer may constitute the biologically relevant functional unit of the enzyme. PMID- 19543811 TI - The effect of the EU tissues and cells directive on bone banking in Denmark: a case study. AB - As a result of the EU Tissues and Cells Directive (2004/23/EC), therapeutic tissue banking is currently being restructured throughout Europe. The stated objectives are to enhance a safe and stable supply of bone and tissue in Europe and to facilitate internal exchange. We conducted an interview study to explore the effect of the Directive on Danish bone banks in terms of (1) organizational restructuring, (2) supply and range of exchange, (3) economic costs. We found that the Directive stimulated extensive re-organization of bone banks with a substantial adjoining workload; that it is doubtful whether it will increase supply and range of exchange; and that the transposition of the Directive is associated with considerable extra cost. Additionally, we found that elements in the documentation of safety were fabricated by surgeons to avoid what was seen as unnecessary questioning of potential donors. PMID- 19543809 TI - Development of physical and mental health summary scores from the patient reported outcomes measurement information system (PROMIS) global items. AB - BACKGROUND: The use of global health items permits an efficient way of gathering general perceptions of health. These items provide useful summary information about health and are predictive of health care utilization and subsequent mortality. METHODS: Analyses of 10 self-reported global health items obtained from an internet survey as part of the Patient-Reported Outcome Measurement Information System (PROMIS) project. We derived summary scores from the global health items. We estimated the associations of the summary scores with the EQ-5D index score and the PROMIS physical function, pain, fatigue, emotional distress, and social health domain scores. RESULTS: Exploratory and confirmatory factor analyses supported a two-factor model. Global physical health (GPH; 4 items on overall physical health, physical function, pain, and fatigue) and global mental health (GMH; 4 items on quality of life, mental health, satisfaction with social activities, and emotional problems) scales were created. The scales had internal consistency reliability coefficients of 0.81 and 0.86, respectively. GPH correlated more strongly with the EQ-5D than did GMH (r = 0.76 vs. 0.59). GPH correlated most strongly with pain impact (r = -0.75) whereas GMH correlated most strongly with depressive symptoms (r = -0.71). CONCLUSIONS: Two dimensions representing physical and mental health underlie the global health items in PROMIS. These global health scales can be used to efficiently summarize physical and mental health in patient-reported outcome studies. PMID- 19543812 TI - Piecing together the elephant: public engagement on nanotechnology challenges. PMID- 19543813 TI - London calling philosophy and engineering: WPE 2008. PMID- 19543816 TI - Powder dissolution method for estimating rotating disk intrinsic dissolution rates of low solubility drugs. AB - PURPOSE: The objective was to investigate the applicability and limitations of a novel approach for measuring intrinsic dissolution rates (IDR) of very small quantities of compounds introduced as powders to buffered solutions and comparing these results to disk IDR obtained using the traditional Wood's apparatus. METHODS: The powder dissolution profiles of 13 model drugs were determined at 37 degrees C in USP buffers at pH 1.2, 4.5, and 6.8, stirred at 100 RPM. As little as 0.06 mg of drug were added to 1 mL buffer media. Drug concentration was measured by an in situ fiber optic UV method. The results were converted to rotating disk IDR values by a novel mathematical procedure. RESULTS: The comparison of the powder-based IDR values to those obtained by traditional Wood's apparatus indicated r(2) = 0.97 (n = 26). CONCLUSION: The results demonstrate that using potentially 10,000-fold less drug material does not sacrifice the quality of the measurement, and lends support to an earlier study that the disk IDR measurement may possibly serve as a surrogate for the BCS solubility classification. PMID- 19543814 TI - Cognitive enhancement: methods, ethics, regulatory challenges. AB - Cognitive enhancement takes many and diverse forms. Various methods of cognitive enhancement have implications for the near future. At the same time, these technologies raise a range of ethical issues. For example, they interact with notions of authenticity, the good life, and the role of medicine in our lives. Present and anticipated methods for cognitive enhancement also create challenges for public policy and regulation. PMID- 19543815 TI - Periostin localizes to cells in normal skin, but is associated with the extracellular matrix during wound repair. AB - Epidermal tissue repair represents a complex series of temporal and dynamic events resulting in wound closure. Matricellular proteins, not normally expressed in quiescent adult tissues, play a pivotal role in wound repair and associated extracellular matrix remodeling by modulating the adhesion, migration, intracellular signaling, and gene expression of inflammatory cells, pericytes, fibroblasts and keratinocytes. Several matricellular proteins show temporal expression during dermal wound repair, but the expression pattern of the recently identified matricellular protein, periostin, has not yet been characterized. The primary aim of this study was to assess whether periostin protein is present in healthy human skin or in pathological remodeling (Nevus). The second aim was to determine if periostin is expressed during dermal wound repair. Using immunohistochemistry, periostin reactivity was detected in the keratinocytes, basal lamina, and dermal fibroblasts in healthy human skin. In pathological nevus samples, periostin was present in the extracellular matrix. In excisional wounds in mice, periostin protein was first detected in the granulation tissue at day 3, with levels peaking at day 7. Periostin protein co-localized with alpha-smooth muscle actin-positive cells and keratinocytes, but not CD68 positive inflammatory cells. We conclude that periostin is normally expressed at the cellular level in human and murine skin, but additionally becomes extracellular during tissue remodeling. Periostin may represent a new therapeutic target for modulating the wound repair process. PMID- 19543817 TI - Measuring cultural identity: validation of a modified Cortes, Rogler and Malgady Bicultural Scale in three ethnic groups in New York. AB - Cultural identity is central to health. Acculturation may be formulated with a bicultural model, assessing in parallel the degree of identification with both the original and the host culture. The Cortes, Rogler and Malgady Bicultural Scale (CRM-BS) is composed of two subscales: "original" culture and "mainstream United States" (US) culture. It was modified into three ethnic versions: Latino, Korean and Chinese. Validation of the CRM-BS was conducted using health professionals and psychiatric patients from the above three ethnic groups and a control sample of mainstream-US (main-US) health professionals in New York City (n = 394). Mean time of completion was 3.7 min and 73% judged it to be easy to use. Strong test-retest reliability correlation coefficients were found (original culture, 0.78; mainstream-US, 0.82). The internal consistency was documented by high Cronbach's alpha values (original culture, 0.88; mainstream-US, 0.80). Factorial analysis revealed two factors, the first one involving all the items of the original culture and the second all of the mainstream-US items. Concerning its discriminant validity, non-main-US subjects scored significantly higher than main-US subjects on the original culture subscale, and vice versa. Construct validity was assessed comparing intergenerational mean scores on both subscales; as generations become older, mean scores for the original culture decreased, while those for the "host" culture increased. Results for each specific ethnic version are also presented. Cutoff scores were calculated to categorize the involvement with the original culture or the host culture, both of them, or neither. PMID- 19543818 TI - The effect of parents' joint work schedules on infants' behavior over the first two years of life: evidence from the ECLSB. AB - We test whether infants living with employed, co-resident parents where at least one parent works a non-standard work shift exhibit significantly more behavior problems than children whose parents both work traditional day shifts. We use a sample of infants living with employed, co-resident parents and two waves of data from the Early Childhood Longitudinal Survey, Birth Cohort (ECLSB) to test whether infants' scores on the Infant-Toddler Symptom Checklist (ITSC) at the second wave (average age of 24.3 months) is affected by parents' shift work at the baseline (average age 10.3 months). Infants with at least one parent who works nonstandard hours have significantly more behavior problems than do infants with parents who both work regular day shifts. This relationship is partly accounted for by shift work's negative association with father-child interaction, marital quality, the frequency of shared family dinners, and parental health, including paternal depression. The results also indicate that shift work has larger effects on children's behavior when mothers, rather than fathers, work nonstandard shifts, and when mothers' day shifts regularly oppose fathers' evenings/night shifts. Policy should focus on giving individuals more choice in their work shift as well as more flexibility in when they start and stop working for the day. Given the importance of mediating factors, we should also focus on ameliorating the negative impacts of shift work when they do arise. This includes addressing issues of employee health and stress, and relationship conflict within couples where one or both partners work a non-standard shift. PMID- 19543820 TI - Synthetic lipopeptide MALP-2 inhibits intracellular growth of Mycobacterium bovis BCG in alveolar macrophages-preliminary data. AB - Alveolar macrophages (AM) are the primary target cell of the lung for inhaled mycobacterial pathogens. We investigated the effect of the synthetic lipopeptide MALP-2 on the interaction between AM from rats and Mycobacterium bovis BCG. AM were infected with M. bovis BCG at a multiplicity of infection (MOI) of 10 and then cultured in medium alone or medium supplemented with either synthetic macrophage activating lipopeptide-2 (MALP-2), or IFN-gamma, or both. Mycobacterial CFU were counted on days 3 and 7 and cell-free supernatants were collected for cytokine measurements. Treatment of macrophages with MALP-2 led to reduced bacterial loads by day 3 and 7 post-infection and at the same time enhanced the release of TNF-alpha, IL-6 und IL-10 compared to non-stimulated, M. bovis BCG-infected AM. Macrophages co-treated with MALP-2 and IFN-gamma or IFN gamma in the absence of MALP-2 limited the growth of M. bovis BCG only by day 3, but not day 7 post-stimulation. Our data show that MALP-2 is effective in decreasing bacterial loads in lung sentinel cells. PMID- 19543819 TI - Effect of salvia miltiorrhizae on pulmonary apoptosis of rats with severe acute pancreatitis or obstructive jaundice. AB - To investigate the effect of apoptosis about Salvia miltiorrhizae injection on the lungs of SAP and OJ rats. Total 288 rats were used for SAP-associated experiments and OJ-associated experiments, respectively. The rats were randomly divided into sham-operated, model control and treated group. According to the difference of time points after operation, the SAP rats in each group were subdivided into 3, 6 and 12 h groups while the OJ rats were divided into 7, 14, 21 and 28 days groups. The pathological changes, expression levels of Bax protein and apoptotic indexes in the lungs of SAP or OJ rats were observed, and the mortality rates of SAP or OJ rats were recorded, respectively. The numbers of dead SAP and OJ rats in treated groups declined. The pathological changes in the lungs of SAP or OJ rats in treated groups were relieved to varying degrees. There was no marked difference in pathological severity scores and the positive staining intensity of Bax protein between treated groups and model control groups (all P > 0.05). Salvia miltiorrhizae has some protective effect on the lungs of rats with SAP or OJ which may be related apoptosis although our results can not find significant difference between treated groups and model control groups. PMID- 19543822 TI - Characterization of the replication-competent porcine endogenous retrovirus class B molecular clone originated from Korean domestic pig. AB - Xenotransplantation from pigs offers an opportunity to resolve the shortage of human organs. The porcine endogenous retrovirus (PERV) cannot be eliminated because of its presence in the germline DNA. Three subgroups of the replication competent PERV (PERV-A, PERV-B, and PERV-C) have been identified in pigs. We constructed a molecular clone of PERV-B from a Korean domestic pig BAC clone containing PERV genomes, and its replication competency was characterized in human cells. The pol region of PERV-B was detected in the genomic DNA of 293T cells transfected with PERV-B (465D1) and in the genomic DNA of 293T cells infected with PERV-B (465D1) viruses. The 293T cells transfected with PERV-B (465D1) were maintained for 140 days. PERV-B (465D1) showed the stationary replication competence with no toxicity in the cell growth. PMID- 19543821 TI - Brief report: inhibitory control of socially relevant stimuli in children with high functioning autism. AB - The current study explored whether inhibitory control deficits in high functioning autism (HFA) emerged when socially relevant stimuli were used and whether arousal level affected the performance. A Go/NoGo paradigm, with socially relevant stimuli and varying presentation rates, was applied in 18 children with HFA (including children with autism or Asperger syndrome) and 22 typically developing children (aged 8-13 years). Children with HFA did not show inhibitory control deficits compared to the control group, but their performance deteriorated in the slow presentation rate condition. Findings were unrelated to children's abilities to recognize emotions. Hence, rather than a core deficit in inhibitory control, low arousal level in response to social stimuli might influence the responses given by children with HFA. PMID- 19543825 TI - The impacts of the HIV/AIDS pandemic and socioeconomic development on the living arrangements of older persons in sub-Saharan Africa: a country-level analysis. AB - This study investigates whether socioeconomic development and the HIV/AIDS pandemic are associated with living arrangement patterns in older persons in 23 sub-Saharan African countries. Country-level aggregate data were taken from previous household surveys and information provided by the United Nations, the World Bank, and the World Health Organization. Results showed that 13.5% of older persons (aged 60 years or over) were living with grandchildren but not adult children (i.e., skipped generation households). Countries higher in HIV/AIDS prevalence had more skipped generation households, and also more older persons living with spouse only and fewer older persons living with other relatives. Countries with higher socioeconomic development had fewer older persons living with children younger than 25 years old and more living with spouse only or with other relatives and unrelated persons. The pandemic and socioeconomic development combine to accelerate the breakdown of the extended family structure so that older persons are less and less likely to reside with, and to receive support from, their children. PMID- 19543824 TI - Antioxidant supplementation of culture medium during embryo development and/or after vitrification-warming; which is the most important? AB - PURPOSE: To determine the most optimal stage for antioxidant supplementation of culture medium to improve developmental competence, cryotolerance and DNA fragmentation of bovine embryos. METHODS: Presumptive zygotes were first cultured in presence or absence of beta-mercaptoethanol (beta-ME), for 8 days. Subsequently, half of the expanded blastocysts developed in both groups were vitrified, warmed within 30 min and post-warming embryos along with their corresponding non-vitrified embryos were cultured for two further days in presence or absence of (100 microM) betaME. RESULTS: For vitrified and non vitrified embryos, the best effect was found when betaME was added from day 1 of in vitro culture in continuation with post-warming culture period. Day 1-8 supplementation significantly increased the rates of cleavage, day 7 and day 8 blastocyst production. For non-vitrified embryos, betaME addition during day 1-8 and/or 9-10 of embryo culture improved both hatching rate and quality of hatched embryos. For vitrified embryos, however, the percentage of DNA-fragmentation (18.5%) was significantly higher (p < or = 0.05) than that of embryos developed in absence of betaME but supplemented with betaME during post-warming period (13.5%). CONCLUSIONS: Exogenous antioxidant increases the chance of embryos, even those of fair-quality, to develop to blastocyst. However, antioxidant inclusion during in vitro embryo development is not sufficient to maintain the redox state of these embryos during the critical period of post-warming embryo culture, and therefore, there should be a surplus source of exogenous antioxidant during post warming embryo culture. PMID- 19543823 TI - Emotional support and gender in people living with HIV: effects on psychological well-being. AB - Current research indicates that emotional support is strongly associated with physical and psychological adjustment in persons living with HIV/AIDS. While gender- differences in health and health behaviors of HIV positive patients are well studied, less is known about how men and women living with HIV/AIDS may differentially perceive and integrate support into their lives, and how it subsequently affects their psychological well-being. This cross-sectional study examines how emotional support received from partners and family/friends and gender explains psychological well-being (i.e., stress, depression, anxiety) in a sample of 409 partnered European HIV positive individuals. We hypothesized that gender would modify the associations between support and psychological well-being such that men would benefit more from partner support whereas women would benefit more from family/friend support. Results revealed that regardless of the source of support, men's well-being was more positively influenced by support than was women's well-being. Women's difficulties in receiving emotional support may have deleterious effects on their psychological well-being. PMID- 19543827 TI - Cyclic AMP inducible early repressor mediates the termination of corticotropin releasing hormone transcription in hypothalamic neurons. AB - Elevations of inducible cAMP early repressor (ICER), the repressor isoform of the cAMP-responsive element modulator (CREM), are associated with protein binding to the corticotrophin releasing hormone (CRH) promoter and termination of CRH transcriptional responses to stress. To determine whether endogenous ICER production represses CRH transcription, we examined the effect of CREM siRNA on forskolin-stimulated ICER formation and CRH transcription in the hypothalamic cell line, 4B, and in primary cultures of hypothalamic neurons. Cotransfection of 4B cells with CREM siRNA and a CRH promoter-driven luciferase reporter gene markedly reduced the induction of ICER by forskolin and potentiated the stimulatory effect of forskolin on CRH promoter activity, compared with cells cotransfected with a nonspecific oligonucleotide. The role of ICER on endogenous CRH expression was studied in primary cultures of hypothalamic neurons by examining the effect of CREM siRNA on forskolin-induced primary transcript (CRH hnRNA) using intronic real-time PCR. As observed during stress in vivo, forskolin stimulated CRH hnRNA was transient, increasing up to 60 min and declining to near basal values by 3 h. Transfection of CREM siRNA reduced forskolin-induced ICER by about 45% 48-h later and partially reversed the declining phase of CRH hnRNA production at 3 h. The data provide evidence that endogenous ICER formation is required for termination of CRH transcription and support the hypothesis that ICER is part of an intracellular feedback mechanism limiting the activation of CRH transcription during stress. PMID- 19543828 TI - Countering countermeasures in the concealed information test using covert respiration measures. AB - The effects of physical and mental countermeasures on the accuracy of the concealed information test (CIT) were examined in a mock crime experiment with 64 participants. To combat countermeasures, two covert respiration measures, hidden in the seat and back of the examination chair, were used in addition to the standard physiological measures (SCR, FPWL, RLL). Some guilty participants were trained to use either physical or mental countermeasures and apply them to distort the outcomes of the CIT. In the second phase of the experiment participants were detached from the standard polygraph devices and examined solely with the two covert measures. Results indicated that physical countermeasures lowered SCR accuracy but had a relatively small effect on the other standard measures. On the other hand, SCR was relatively resistant to mental countermeasures. Both covert measures were resistant to physical countermeasures in the polygraph phase. When the standard devices were removed, the covert seat measure was effective in the no countermeasure and in the mental countermeasure conditions but not when physical countermeasures were applied. The back measure was entirely ineffective. PMID- 19543826 TI - MDMA (Ecstasy) decreases the number of neurons and stem cells in embryonic cortical cultures. AB - Ecstasy, 3,4-methylenedioxymetamphetamine (MDMA), is a recreational drug used among adolescents, including young pregnant women. MDMA passes the placental barrier and may therefore influence fetal development. The aim was to investigate the direct effect of MDMA on cortical cells using dissociated CNS cortex of rat embryos, E17. The primary culture was exposed to a single dose of MDMA and collected 5 days later. MDMA caused a dramatic, dose-dependent (100 and 400 microM) decrease in nestin-positive stem cell density, as well as a significant reduction (400 microM) in NeuN-positive cells. By qPCR, MDMA (200 microM) caused a significant decrease in mRNA expression of the 5HT3 receptor, dopamine D(1) receptor, and glutamate transporter EAAT2-1, as well as an increase in mRNA levels of the NMDA NR1 receptor subunit and the 5HT(1A) receptor. In conclusion, MDMA caused a marked reduction in stem cells and neurons in embryonic cortical primary cell cultures, which was accompanied by changes in mRNA expression of specific receptors and transporters for glutamatergic and monoaminergic neurotransmitters. PMID- 19543830 TI - Reversal of the glycolytic phenotype by dichloroacetate inhibits metastatic breast cancer cell growth in vitro and in vivo. AB - The glycolytic phenotype is a widespread phenomenon in solid cancer forms, including breast cancer. Dichloroacetate (DCA) has recently been proposed as a novel and relatively non-toxic anti-cancer agent that can reverse the glycolytic phenotype in cancer cells through the inhibition of pyruvate dehydrogenase kinase. We have examined the effect of DCA against breast cancer cells, including in a highly metastatic in vivo model. The growth of several breast cancer cell lines was found to be inhibited by DCA in vitro. Further examination of 13762 MAT rat mammary adenocarcinoma cells found that reversal of the glycolytic phenotype by DCA correlated with the inhibition of proliferation without any increase in cell death. This was despite a small but significant increase in caspase 3/7 activity, which may sensitize cancer cells to other apoptotic triggers. In vivo, DCA caused a 58% reduction in the number of lung metastases observed macroscopically after injection of 13762 MAT cells into the tail vein of rats (P = 0.0001, n > or = 9 per group). These results demonstrate that DCA has anti proliferative properties in addition to pro-apoptotic properties, and can be effective against highly metastatic disease in vivo, highlighting its potential for clinical use. PMID- 19543829 TI - Telomere length in blood cells and breast cancer risk: investigations in two case control studies. AB - Telomere dysfunction, which leads to genomic instability, is hypothesized to play a causal role in the development of breast cancer. However, the few epidemiologic studies that assessed the relationship between telomere length in blood cells and breast cancer risk have been inconsistent. We conducted two case-control studies to further understand the role of telomere length and breast cancer risk. Overall telomere lengths were measured by telomere quantitative fluorescent in situ hybridization (TQ-FISH) and telomere quantitative real-time PCR (TQ-PCR). The associations between telomere length in blood leukocytes and risk of breast cancer were examined in two breast cancer case-control studies that were conducted at Roswell Park Cancer Institute (RPCI) and Lombardi Comprehensive Cancer Center (LCCC). Using the 50th percentile value in controls as a cut point, women who had shorter telomere length were not at significantly increased risk of breast cancer compared with women who had longer telomere length in the RPCI study (odds ratio [OR] = 1.34, 95% confidence interval [CI] = 0.84-2.12), in the LCCC study (OR = 1.18, 95% CI = 0.73-1.91), or in the combined RPCI and LCCC studies (OR = 1.23, 95% CI = 0.89-1.71). There was no significant dose-response relationship across quartiles of telomere length and no significant difference when comparing women in the lowest to highest quartile of telomere length. Overall telomere length in blood leukocytes was not significantly associated with the risk of breast cancer. PMID- 19543832 TI - A magnetic approach to decrease stent graft endoleak: ex-vivo validation. AB - Stent graft endoleak is a major problem in endovascular aneurysm repair (EVAR). Endoleak occurs after EVAR and may lead to aneurysm rupture, acute vessel thrombosis or occlusion. This study presents a novel design to potentially reduce endoleak with use of magnets. A ferromagnetic stent is deployed into the vessel lumen, and two external flexible magnetic rings are used to clamp on the proximal and distal necks of the stent. The rings impose epivascular magnetic pressure on the vessel wall to prevent the vessel wall from separating from the stent under elevated blood pressure. The geometry and magnetic properties of the stent and rings were designed to produce sufficient pressure, without overly compressing the vessel wall. Feasibility of this design was demonstrated with in vitro experiments using porcine abdominal aortas. The experiments showed that magnetic ring significantly improved the seal between the stent and vessel wall with use of moderate-sized stent. For aortas subjected to physiological axial stretch, rings that generate magnetic pressure of about 45 mmHg were found sufficient to prevent endoleak at pressure of 140 mmHg. Evaluation of this design in an in vivo animal model of aortic aneurysm is warranted. PMID- 19543831 TI - Decrease of Tau hyperphosphorylation by 17beta estradiol requires sphingosine kinase in a glutamate toxicity model. AB - Several studies have linked estrogens with sphingosine kinase (SphK) activity, enzyme responsible of sphingosine-1-phosphate synthesis (S-1P), however their possible interaction in the nervous system is not documented yet. In the present study, we developed a glutamate toxicity model in SH-SY5Y cells to evaluate the possible effect of the inhibition of SphK activity on the protective capability of 17beta-estradiol (E2). Glutamate induced cytoskeletal actin changes associated to cytotoxic stress, significant increase of apoptotic-like nuclear fragmentation, Tau hyperphosphorylation and increase of p25/p35 cleavage. These effects were prevented by E2 pre-treatment during 24 h. Although the inhibition of SphK did not block this protective effect, significantly increased Tau hyperphosphorylation by glutamate, in a way that was not reverted by E2. Our results suggest that the decrease of glutamate-induced Tau hyperphosphorylation by 17beta-estradiol requires SphK. PMID- 19543833 TI - Conceptual competence as a component of second language fluency. AB - In this article we argue that conceptual competence should be seen as a component of second language (L2) communicative competence. Abstract concepts are highly expressed by means of metaphors, metonymies, idioms and other types of figurative language. In literature it is suggested that knowledge and appropriate use of these lexical segments are closely related to L2 mastery and therefore conceptual instruction is expected to facilitate L2 learning. To test the relationship between conceptual and L2 competence we conducted an experiment in which Modern Greek learners were encouraged to express their views on the concept of happiness. The results showed their weak performance in conveying their ideas in a coherent and acceptable manner. PMID- 19543834 TI - Earthquake-prone cities. PMID- 19543836 TI - Identifying hidden sexual bridging communities in Chicago. AB - Bridge populations can play a central role in the spread of human immunodeficiency virus (HIV) by providing transmission links between higher and lower prevalence populations. While social network methods are well suited to the study of bridge populations, analyses tend to focus on dyads (i.e., risk between drug and/or sex partners) and ignore bridges between distinct subpopulations. This study takes initial steps toward moving the analysis of sexual network linkages beyond individual and risk group levels to a community level in which Chicago's 77 community areas are examined as subpopulations for the purpose of identifying potential bridging communities. Of particular interest are "hidden" bridging communities; that is, areas with above-average levels of sexual ties with other areas but whose below-average AIDS prevalence may hide their potential importance for HIV prevention. Data for this analysis came from the first wave of recruiting at the Chicago Sexual Acquisition and Transmission of HIV Cooperative Agreement Program site. Between August 2005 through October 2006, respondent driven sampling was used to recruit users of heroin, cocaine, or methamphetamine, men who have sex with men regardless of drug use, the sex partners of these two groups, and sex partners of the sex partners. In this cross-sectional study of the sexual transmission of HIV, participants completed a network-focused computer assisted self-administered interview, which included questions about the geographic locations of sexual contacts with up to six recent partners. Bridging scores for each area were determined using a matrix representing Chicago's 77 community areas and were assessed using two measures: non-redundant ties and flow betweenness. Bridging measures and acquired immunodeficiency syndrome (AIDS) case prevalence rates were plotted for each community area on charts representing four conditions: below-average bridging and AIDS prevalence, below-average bridging and above-average AIDS prevalence, above-average bridging and AIDS prevalence, and above-average bridging and below-average AIDS prevalence (hidden bridgers). The majority of the 1,068 study participants were male (63%), African American (74%), and very poor, and the median age was 44 years. Most (85%) were sexually active, and 725 provided useable geographic information regarding 1,420 sexual partnerships that involved 57 Chicago community areas. Eight community areas met or came close to meeting the definition of hidden bridgers. Six areas were near the city's periphery, and all eight areas likely had high inflows or outflows of low-income persons displaced by gentrification. The results suggest that further research on this method is warranted, and we propose a means for public health officials in other cities to duplicate the analysis. PMID- 19543835 TI - A multilevel analysis of social ties and social cohesion among Latinos and their neighborhoods: results from Chicago. AB - Research suggests that, among Latinos, there are health benefits associated with living in a neighborhood populated with coethnics. While social networks and social cohesion are the proposed explanation for the salubrious effect and are assumed to be characteristics of Latino immigrant enclaves, evidence for this is limited. We used multilevel regression to test the relative contribution of individual race/ethnicity and neighborhood concentration of Mexican Americans as predictors of social networks and social cohesion. After accounting for personal characteristics, we found a negative association between neighborhood concentration of Mexican Americans and social cohesion. Among Latinos, living in a neighborhood with increased coethnics was associated with increased social ties. Compared to non-Latino whites, Mexican Americans reported more social ties but lower social cohesion. Contrary to the assumption that Mexican immigrant enclaves beget social cohesion, we did not find this to be true in Chicago neighborhoods. PMID- 19543837 TI - Bridging sexual boundaries: men who have sex with men and women in a street-based sample in Los Angeles. AB - The purpose of the study was to determine the potential contribution of bisexual men to the spread of HIV in Los Angeles. We compare the characteristics and behaviors of men who have sex with men and women (MSMW) to men who have sex with only women (MSW) and men who have sex with only men (MSM) in Los Angeles. Men (N = 1,125) who participated in one of the two waves of data collection from 2005 to 2007 at the Los Angeles site for NIDA's Sexual Acquisition and Transmission of HIV-Cooperative Agreement Program were recruited using Respondent Driven Sampling. Participants completed Audio Computer Assisted Self Interviews and received oral HIV rapid testing with confirmatory blood test by Western Blot and provided urine specimens for detection of recent powder cocaine, crack cocaine, methamphetamine, or heroin use. MSM, MSW, or MSMW were defined by the gender of whom they reported sex with in the past 6 months. Chi-square tests and ANOVAs were used to test independence between these groups and demographic characteristics, substance use, and sexual behaviors. We fit generalized linear random intercept models to predict sexual risk behaviors at the partner level. Men were mostly of low income, unemployed, and minority, with many being homeless; 66% had been to jail or prison, 29% had ever injected drugs, and 25% had used methamphetamine in the past 30 days. The sample had high HIV prevalence: 12% of MSMW, 65% of MSM, and 4% of MSW. MSMW were behaviorally between MSW and MSM, except that more MSMW practiced sex for trade (both receiving and giving), and more MSMW had partners who are drug users than MSW. Generalized linear random intercept models included a partner-level predictor with four partner groups: MSM, MSMW-male partners, MSMW-female partners, and MSW. The following were significantly associated with unprotected anal intercourse (UAI): MSW (AOR 0.15, 95% CI 0.08, 0.27), MSMW-female partners (AOR 0.4, 95% CI 0.27, 0.61), HIV positive partners (AOR 2.03, 95% CI 1.31, 3.13), and being homeless (AOR 1.37, 95% CI 1.01, 1.86). The factors associated with giving money or drugs for sex were MSMW-female partners (AOR 1.70, 95% CI 1.09, 2.65), unknown HIV status partners (AOR 1.72, 95% CI 1.29, 2.30), being older (AOR 1.02, 95% CI 1.00, 1.04), history of incarceration (AOR 1.64, 95% CI 1.17, 2.29), and being homeless (AOR 1.73, 95% CI 1.27, 2.36). The following were associated with receiving money or drugs for sex: MSW (AOR 0.53, 95% CI 0.32, 0.89), African American (AOR 2.42, 95% CI 1.56, 3.76), Hispanic (AOR 1.85, 95% CI 1.12, 3.05), history of incarceration (AOR 1.44, 95% CI 1.04, 2.01), history of injecting drugs (AOR 1.57, 95% CI 1.13, 2.19), and had been recently homeless (AOR 2.14, 95% CI 1.57, 2.94). While overall HIV-positive MSM had more UAI with partners of any HIV status than MSMW with either partner gender, among HIV-positive MSMW, more had UAI with HIV-negative and HIV status unknown female partners than male partners. Findings highlight the interconnectedness of sexual and drug networks in this sample of men-as most have partners who use drugs and they use drugs themselves. We find a concentration of risk that occurs particularly among impoverished minorities-where many men use drugs, trade sex, and have sex with either gender. Findings also suggest an embedded core group of drug-using MSMW who may not so much contribute to spreading the HIV epidemic to the general population, but driven by their pressing need for drugs and money, concentrate the epidemic among men and women like themselves who have few resources. PMID- 19543838 TI - Evaluation of antineoplastic activity of extracellular asparaginase produced by isolated Bacillus circulans. AB - L-Asparaginase is an important component in the treatment of acute lymphoblastic leukemia in children. Its antineoplastic activity toward malignant cells is due to their characteristic nature in slow synthesis of L-asparagine (Asn), which causes starvation for this amino acid, while normal cells are protected from Asn starvation due to their ability to produce this amino acid. The relative selectivity with regard to the metabolism of malignant cells forces to look for novel asparaginase with little glutaminase-producing systems compared to existing enzyme. In this investigation, the role of the extracellular asparaginase enzyme produced by an isolated bacterial strain was studied. Biochemical characterization denoted that this isolated bacterial strain belongs to the Bacillus circulans species. The strain was tested for L-asparaginase production, and it was observed that, under an optimized environment, this isolate produces a maximum of 85 IU ml(-1) within 24-h incubation. This enzyme showed less (60%) glutaminase activity compared to commercial Erwinia sp. L-asparaginase. The partially purified enzyme showed an approximate molecular weight of 140 kDa. This enzyme potency in terms of antineoplastic activity was analyzed against the cancer cells, CCRF-CEM. Flow cytometry experiments indicated an increase of sub G1 cell population when the cells were treated with L-asparaginase. PMID- 19543839 TI - P53 Arg72Pro polymorphism in gastric cancer patients. AB - INTRODUCTION: Polymorphism in codon 72, exon 4 of p53 may alter apoptosis and cancer progression. PATIENTS AND METHODS: P53 Arg72Pro genotype was assessed by PCR from 84 gastric cancer patients and 185 controls. The control group was comparable in sex, race, age, smoking, and alcohol intake to the cancer group. RESULTS AND DISCUSSION: There was no difference among the frequency of the alleles or genotypes between the groups. P53 Pro/Pro was associated to a lower risk of metastatic disease (p = 0.02) but not to lymph nodes metastasis or worst prognosis. Arg/Arg or Arg/Pro genotype may be associated to metastatic disease. PMID- 19543840 TI - Thyroid hormones alterations during acute liver failure: possible underlying mechanisms and consequences. AB - Thyroid hormones are now recognized to change in different disease states with important consequences on severity and prognosis of disease. However, little is known about thyroid hormones' alterations in acute liver failure (ALF). To study the changes in thyroid hormones and cardiac thyroid receptors during ALF, we subjected seven female pigs to surgical liver devascularization. Liver function biochemical markers, thyroid hormones, endogenous opioids, malondialdehyde (MDA), and interleukins 1 and 6 were measured in serum for 24 h postoperatively. Heart biopsies were harvested at the end of the experiment. Baseline heart biopsies were taken from five additional animals. Serum thyroxin (T(4)) and triiodothyronine (T(3)) levels markedly decreased, whereas free-triiodothyronine and thyroxin-stimulating hormone levels did not change. T(4) and T(3) levels correlated with the degree of liver failure and with MDA and interleukin-6 levels. Beta-endorphin levels initially increased, whereas levels of leucine enkephalin did not change. Thyroid hormone receptor-alpha1 protein expression in the heart decreased 1.6-fold after ALF, whereas myocardial myosin isoform expression remained unchanged. The downregulation of T(4) and T(3) levels during ALF seems to correlate well with the severity of disease. This downregulation related to inflammation and oxidative stress and resulted in changes in myocardial thyroid receptors. PMID- 19543841 TI - Severe osteomalacia presenting with multiple vertebral fractures: a case report and review of the literature. AB - Adequate exposure to sunlight and fortification of dairy products with vitamin D have eliminated vitamin D deficiency secondary to inadequate endogenous production or nutrition in the majority of countries. Insufficient vitamin D intake secondary to using unfortified foods and social customs (such as avoiding sun exposure), however, contribute to the development of disease. Poor diet, a lack of sun exposure, and the age related decline in the dermal synthesis of 7 dehydrocholesterol are among the factors that predispose to vitamin D deficiency and consequent bone disease. Here, we present a case of severe osteomalacia presenting with multiple vertebral fractures due to poor diet and a lack of exposure to sunlight. PMID- 19543842 TI - Role of mesenchymal stem cells in immunological rejection of organ transplantation. AB - The discovery that Mesenchymal Stem Cells (MSCs) can strongly inhibit T cell proliferation in vitro and in vivo and exert similar inhibitory effects on B cells, dendritic cells, and natural killer cells has highlighted the potential for clinical translation of these cells as a new class of stem cell therapy for autoimmune disease, organ transplantation and treatment of graft-versus-host disease (GVHD). Even though the mechanism underlying these immunosuppressive effects of MSCs has not been clearly defined, their immunosuppressive properties are already being exploited in the clinical setting. Most of these early clinical studies are investigating the effect of MSCs in suppressing GVHD after allogenenic hematopoietic stem cell transplantation (HSCT). Additional studies, mostly in animal models, are being conducted in solid organ transplantation, such as: heart, renal, liver and skin. While the early results of these studies are conflicting, the potential for clinical benefit remains high and further studies are warranted in order to discover the best methods and settings for consistent clinical results. MSCs have opened a series of opportunities for researchers in the areas of transplantation and autoimmune disease. While it is important not to overestimate the potential therapeutic effects of MSCs, and well-designed preclinical trials should be done before clinical use. PMID- 19543843 TI - Better off in the community? A 5-year follow up study of long-term psychiatric patients discharged into the community. AB - BACKGROUND: The quality of life of long-term psychiatric inpatients relocated to the community was investigated in this study. The aim was to investigate what changes, if any occurred, on standardised quality of life related instruments between discharge from hospital and at 1 year after discharge into the community. We were also interested to see if these changes continued 5 years after discharge into the community. METHOD: 87 long-stay psychiatric patients were enrolled in the study. Each patient was assessed on four standardised assessment instruments designed to assess their attitudes towards community living and level of functioning in the community. RESULTS: Patients reported being satisfied in their new community environment. They showed improvements in their level of self-care and social functioning after 1 year in the community. These improvements were not maintained in their fifth year in the community. In addition, there were no improvements in patient's domestic skills, community skills or activity and social relations levels. Weekly occupation levels increased after 5 years in the community and their level of interests in things increased over the first year but not after 5 years in the community. CONCLUSIONS: This study adds to the previous work carried out on patients discharged from large psychiatric hospitals into the community. Patients expressed a desire to continue to live in the community and while they showed improvements in self care and social functioning in the first year following discharge, these improvements were not sustained after 5 years in the community. Further training is needed for staff in the community residences so that patients can achieve their maximum potential. PMID- 19543845 TI - Shorter unipedal standing time and lower bone mineral density in women with distal radius fractures. AB - Unipedal standing time was shorter and bone mineral density was lower in Japanese women aged 50 years and over with low-energy distal radius fractures resulting from falls than those in age-matched community-dwelling Japanese women without distal radius fractures. INTRODUCTION: The aim of this study was to compare unipedal standing time and bone mineral density (BMD) of women >or=50 years of age with distal radius fractures with those of age-matched women without fractures. METHODS: Fracture group was 54 Japanese women with low-energy distal radius fractures resulting from fall. Non-fracture group was 52 community dwelling Japanese women without fractures. Unipedal standing time and BMD were measured. RESULTS: There were no significant differences in age and body mass index between the two groups. The percentage of women with unipedal standing time <15 s was 44.4% in the fracture group and 13.5% in the non-fracture group, while the respective frequencies for >120 s were 20.4% and 50.0%. The T-score of BMD was significantly lower in the fracture than non-fracture group. Logistic regression analysis identified unipedal standing time <15 s and T-score <70% as significant factors associated with distal radius fractures. Notably, T-score <70% was significant in subjects <65 years, and unipedal standing time <15 s was significant in those >or=65 years. CONCLUSION: Unipedal standing time was shorter and BMD was lower in women >or=50 years of age with distal radius fractures than those in age-matched women without fractures. PMID- 19543844 TI - The prevalence of personality disorder, its comorbidity with mental state disorders, and its clinical significance in community mental health teams. AB - BACKGROUND: Personality status is seldom assessed in community mental health teams except at a rudimentary level. This study challenges the assumption that this policy is either prudent or wise. AIMS: To measure the prevalence of personality disorder within community mental health teams and to investigate its relationship to mental state disorders and overall pathology. METHOD: A cross sectional survey of 2,528 of 2,567 psychiatric patients (98.5%) managed by community mental health teams in four urban settings in the UK in which diagnoses of personality and mental state pathology were assessed separately. Of these, a sample of 400 was interviewed, with a 70.5% completion rate for more in depth information. RESULTS: In total, 40% of all patients in secondary care suffered from at least one personality disorder. Regression modelling showed personality pathology accounted for a greater degree of global psychopathology than psychosis, alcohol or drug dependence, but was associated with anxiety disorders. CONCLUSION: Comorbid personality pathology contributes greatly to overall psychopathology in secondary psychiatric care. It should be both recognised and managed. PMID- 19543847 TI - Left ventricular accessory chamber: a case report and review of the literature. AB - Congenital left ventricular out-pouching, consisting of left ventricular aneurysms, left ventricle diverticulae, and double-chambered left ventricle, is a rare cardiac malformation. Criteria to differentiate between left ventricular aneurysm and diverticulum are defined. Reports of left ventricular accessory chamber in the medical literature are rare. In this article, we present a case of left ventricular accessory chamber diagnosed during the late prenatal period. Review of the literature, imaging modalities used for differentiation of the nature of this lesion, and management plan are discussed. PMID- 19543846 TI - High interleukin-6 and tumor necrosis factor-alpha serum levels in hepatitis C infection associated or not with mixed cryoglobulinemia. AB - The objective of this study is to evaluate serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-a) in a series of patients with hepatitis C virus (HCV)-related mixed cryoglobulinemia (HCV-MC) and to correlate these parameters with the clinical features of the disease. Serum IL-6 and TNF-a were assayed in 61 patients with HCV-MC, in 61 sex- and age-matched patients with HCV chronic hepatitis without cryoglobulinemia (HCV+), and in 61 sex- and age-matched healthy controls. HCV-MC patients showed significantly higher mean IL-6 levels than controls (p=0.005) or HCV+ patients (p = 0.02). Moreover, IL-6 was increased in cryoglobulinemic patients with active vasculitis, even if the statistical significance was not reached (p=0.056). Serum TNF-a levels were significantly higher in HCV-MC than in HCV+ or in controls (p<0.01). The study demonstrates high IL-6 and TNF-a serum levels in HCV-MC patients; moreover, IL-6 levels tended to be higher in HCV-MC patients in presence of active vasculitis. PMID- 19543848 TI - Is the current therapeutic armamentarium in diabetes enough to control the epidemic and its consequences? What are the current shortcomings? AB - The prevalence of diabetes is expected to rise together with an increase in morbidity and a reduction in life expectancy. A leading cause of death is cardiovascular disease, and hypertension and diabetes are additive risk factors for this complication. Selected treatment options should neither increase cardiovascular risk in patients with diabetes, nor increase risk of hyperglycaemia in patients with hypertension. The efficacy of present antihyperglycaemic agents is limited and new therapies, such as incretin-targeted agents, are under development. Even though most patients do not achieve glycated haemoglobin targets, trial data show that such interventions reduce the incidence of macrovascular events; however, intensive lowering may be detrimental in patients with existing cardiovascular disease. Currently available oral drugs do not address the key driver of type 2 diabetes--loss of functional beta-cell mass. In the future, new oral treatments must improve this, whilst providing durable blood glucose control and long-term tolerability. PMID- 19543849 TI - Genomic sequence of a Japanese encephalitis virus isolate from southern China. AB - We determined the complete nucleotide sequence of a Japanese encephalitis virus (JEV) isolate (designated SH17M-2007) from a pool of Culex tritaeniorhynchus collected in southern China in 2007. The genome consisted of 10,965 nucleotides and included a single open reading frame (10,296 nucleotides) that encodes a 3,432-amino-acid polyprotein. The SH17M-2007 had 97.3 to 98.4% nucleotide identity with two Korean strains (KV1899, K94P05) and two Japanese strains (Ishikawa, JEV/sw/Mie/40/2004), but only 88.8% identity with the Chinese vaccine strain SA14-14-2. Five unique amino acid substitutions including one in the envelope (E) protein (Glu(E-306)-Lys) were found in the SH17M-2007 strain. Phylogenetic relationships based on the full-length nucleotide sequences were similar to those based on the E gene. PMID- 19543850 TI - Crystal structures of g-type lysozyme from Atlantic cod shed new light on substrate binding and the catalytic mechanism. AB - Crystal structures of Atlantic cod lysozyme have been solved with and without ligand bound in the active site to 1.7 and 1.9 A resolution, respectively. The structures reveal the presence of NAG in the substrate binding sites at both sides of the catalytic Glu73, hence allowing the first crystallographic description of the goose-type (g-type) lysozyme E-G binding sites. In addition, two aspartic acid residues suggested to participate in catalysis (Asp101 and Asp90) were mutated to alanine. Muramidase activity data for two single mutants and one double mutant demonstrates that both residues are involved in catalysis, but Asp101 is the more critical of the two. The structures and activity data suggest that a water molecule is the nucleophile completing the catalytic reaction, and the roles of the aspartic acids are to ensure proper positioning of the catalytic water. PMID- 19543852 TI - Proteomic analysis reveals significant elevation of heat shock protein 70 in patients with chronic heart failure due to arrhythmogenic right ventricular cardiomyopathy. AB - As proteins are the ultimate biological determinants of phenotype of disease, we screened altered proteins associated with heart failure due to arrhythmogenic right ventricular cardiomyopathy (ARVC) to identify biomarkers potential for rapid diagnosis of heart failure. By 2-dimensional gel electrophoresis and mass spectrometry, we identified five commonly altered proteins with more than 1.5 fold changes in eight ARVC failing hearts using eight non-failing hearts as reference. Noticeably, one of the altered proteins, heat shock protein 70 (HSP70), was increased by 1.64 fold in ARVC failing hearts compared with non failing hearts. The increase of cardiac HSP70 was further validated by Western blot, immunochemistry, and enzyme-linked immunosorbent assay (ELISA) in failing hearts due to not only ARVC, but also dilated (DCM, n = 18) and ischemic cardiomyopathy (ICM, n = 8). Serum HSP70 was also observed to be significantly increased in heart failure patients derived from the three forms of cardiomyopathies. In addition, we observed hypoxia/serum depletion stimulation induced significantly elevation of intracellular and extracellular HSP70 in cultured neonatal rat cardiomyocytes. For the first time to our knowledge, we revealed and clearly demonstrated significant up-regulation of cardiac and serum HSP70 in ARVC heart failure patients. Our results indicate that elevated HSP70 is the common feature of heart failure due to ARVC, DCM, and ICM, which suggests that HSP70 may be used as a biomarker for the presence of heart failure due to cardiomyopathies of different etiologies and may hold diagnostic/prognostic potential in clinical practice. PMID- 19543851 TI - Atypical choroid plexus papilloma: clinical experience in the CPT-SIOP-2000 study. AB - Atypical choroid plexus papilloma (APP) represents a novel intermediate-grade subtype of choroid plexus tumor (CPT), the clinical outcome of which has not been described yet. We present the first analysis of a group of APP patients enrolled in the ongoing CPT-SIOP-2000 study of CPTs. A worldwide registration and a randomized trial for those patients who require chemotherapy started in 2000. For APP, maximal surgical resection was recommended. After surgery, patients who had undergone complete resection were observed, whereas patients with incompletely resected or metastasized APP were treated with six chemotherapy courses (etoposide and vincristine, combined with either carboplatin or cyclophosphamide). Risk-adapted radiotherapy was given only to patients older than 3 years of age. Of the 106 patients with a centrally confirmed CPT histology, 30 had APP, 42 CPP and 34 CPC. APP patients were significantly younger (median = 0.7 years) than patients with CPP or CPC (both medians = 2.3 years). Complete resection was achieved in 68 (64%) patients (79% in CPP, 63% in APP, and 47% in CPC). Metastases were present at diagnosis in 17% of APP patients, 5% of CPP patients, and 21% of CPC patients. All nine APP patients who received postoperative chemotherapy showed an early response after two cycles: two had complete remission, four had partial response, and three had stable disease. In the observation group of 15 patients, one event was seen, and all patients were alive. In the treatment group, one patient with a metastasized tumor and incompletely resected APP died. While APP was defined histologically, median percentages of both the Ki-67/MIB-1 proliferation marker and the p53 tumor suppressor protein increased across the three histological subtypes (from CPP to APP and then CPC), suggesting that the subtypes comprise an ordinal categorization of increasingly severe CPT tumors. This ordering was reiterated by clinical outcome in the 92 patients treated per the study protocol, with 5-year EFS rates of 92% in 39 CPP patients, 83% in 24 APP patients, and 28% in 29 CPC patients. A similar ordering was seen when all 106 patients were evaluated for EFS. APP responded favorably to chemotherapy. The intermediate position of APP between CPP and CPC was supported by the clinical data. PMID- 19543853 TI - Endothelial dysfunction in the early- and late-stage type-2 diabetic Goto Kakizaki rat aorta. AB - As there are increasing evidences that human diabetes induces cardiovascular dysfunction, we investigated the type-2 diabetes-induced endothelial dysfunction in the early and late-stage Goto-Kakizaki (GK) rat aorta. We performed organ bath studies, and examined the changes in expression levels of muscarinic M(3) receptor, endothelial, inducible, and neuronal nitric oxide synthase (eNOS, iNOS, and nNOS, respectively) mRNAs in the rat aorta utilizing real-time polymerase chain reaction in 12-week-old and 70-week-old GK rats as well as in age-matched Wistar rats. In the 12-week-old GK rat aorta, a significant increase in norepinephrine-induced contraction and a significant decrease in acetylcholine induced relaxation as well as significant increases in expression levels of muscarinic M(3) receptor and eNOS and a significant decrease in nNOS mRNAs were observed compared to age-matched controls. In the older GK rat aorta, significant decreases in acetylcholine- and nitroglycerine-induced relaxations as well as significant decreases in the expression levels of muscarinic M(3) receptor, eNOS, iNOS, and nNOS mRNAs were observed compared to those in the younger GK rats. In contrast, although significant decreases in acetylcholine and nitroglycerine induced relaxations were observed, the expression levels of muscarinic M(3) receptor, eNOS, iNOS, and nNOS mRNAs in the older Wistar rats aorta were unchanged, increased, increased and decreased, respectively, compared to the younger Wistar rat aorta. These results indicate that endothelial dysfunction in the rat aorta progresses with age and development of diabetes condition, and that decreased relaxations in the late-stage rat aorta may be due to these alterations. PMID- 19543854 TI - Specific biofunctional performances of the hydroxyapatite-sodium maleate copolymer hybrid coating nanostructures evaluated by in vitro studies. AB - The nanohybrid structures consisting of hydroxyapatite (HA) and sodium maleate vinyl acetate copolymer (MP) deposited by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique on Ti surfaces were investigated for specific biological qualities required in bone implantology. The data from in vitro studies demonstrated that human primary osteoblasts (OBs) firmly adhered to Ti coated with HA-MP as indicated by cytoskeleton and vinculin dynamics. OBs spread onto biomaterial surface and formed groups of cells which during their biosynthetic activity expressed OB phenotype specific markers (collagen and non collagenous proteins) and underwent controlled proliferation. PMID- 19543855 TI - Integrity of a recombinant hemagglutinin protein of an avian influenza virus. AB - An open reading frame representing cDNA from a hemagglutinin (HA) encoding gene of a low pathogenic avian influenza virus (AIV) subtype H10N7 was cloned in the pNMT1-TOPO vector under the control of thiamine response promoter. This construct was designated as pNMT1-HA. The pNMT1-HA construct was transformed into Schizosaccharomyces pombe for expression of HA antigen. The correct expression of recombinant HA protein was confirmed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. The level of expression of recombinant HA protein was approximately 0.2% of total soluble protein. Purified yeast-derived recombinant HA protein showed hemagglutination activity. The 2-D and 3-D scanning images of recombinant HA protein were observed with an atomic force microscope (AFM). The structural integrity of the HA protein under AFM and hemagglutination activity provided support that the recombinant HA protein may be suitable for development of AIV subunit vaccine for mass administration to poultry. PMID- 19543856 TI - Gene-specific silencing induced by parallel complementary RNA in Pseudomonas aeruginosa. AB - To investigate whether parallel complementary RNA (pRNA) could induce gene specific silencing in Pseudomonas aeruginosa, pRNA of the mexA gene was expressed in it. Compared to the control strains, the strain expressing pRNA of mexA showed a 50% decrease in minimum inhibitory concentrations (MICs) of several antimicrobial agents and a twofold increase in the initial accumulation rate of ethidium bromide, all of which are substrates of the MexAB-OprM efflux pump. These results suggest that gene-specific silencing was induced by pRNA. This is the first time that such a route for gene silencing has been reported in a bacterium other than Escherichia coli. Gene-specific silencing induced by pRNA may be useful as a novel biotechnology tool for gene regulation in prokaryotes. PMID- 19543858 TI - Genetic transformation of cork oak (Quercus suber L.) for herbicide resistance. AB - The bar gene was introduced into the cork oak genome. Cork oak embryogenic masses were transformed using the Agrobacterium strain AGL1 which carried the plasmid pBINUbiBar. This vector harbours the genes, nptII and bar, the latter under control of the maize ubiquitin promoter. The transgenic embryogenic lines were cryopreserved. Varying activities of phosphinothricin acetyl transferase were detected among the lines, which carried 1-4 copies of the insert. Molecular and biochemical assays confirmed the stability and expression of the transgenes 3 months after thawing the cultures. These results demonstrate genetic engineering of herbicide tolerance in Quercus spp. PMID- 19543861 TI - [A 39-year-old patient with maculopapular rash, recurrent fever, and arthralgia]. AB - Adult-onset Still's disease is a rare disorder of unknown etiology. We report the case of a 39-year-old patient who showed the characteristic symptoms: recurrent attacks of fever, arthralgia, maculopapular rash, sore throat, and lymphadenopathy. After the possibility of an infectious or paraneoplastic process was excluded and the laboratory findings were evaluated (increased C-reactive protein, liver values, and ferritin level), the diagnosis was established according to the criteria of Yamaguchi. Therapy with steroids and nonsteroidal anti-inflammatory drugs was started successfully. PMID- 19543860 TI - [Strategy for avoidance of negative appendectomies]. AB - Appendectomy is the most commonly performed emergency surgical procedure. Even in the era of laparoscopic surgery and modern computed tomography, the rate of negative appendectomies, defined as the removal of a non-inflamed appendix, remains high (10-15%). The general problem and incidence, as well as the influence of modern diagnostic modalities on the rate of negative appendectomies are of particular clinical relevance. Several clinical scoring systems have been developed, but they did not find their way into the daily clinical routine. A proposed diagnostic algorithm could support further efforts to reduce the rate of negative appendectomies. The risk of overtreatment in the reduction of the negative appendectomy rate might potentially lead to acceptance of a higher perforation rate. PMID- 19543859 TI - Mycobacterium tuberculosis, autoimmunity, and vitamin D. AB - Mycobacterium tuberculosis (TB) is a major cause of morbidity and mortality worldwide. Current anti-TB chemotherapies, although effective, are associated with side effects and are limited in treating drug-resistant strands. Autoimmune diseases are a leading cause of morbidity and mortality, with a growing mass of evidence implicating infections (e.g., TB) as their triggers. The burden of TB might further increase by reactivation threats hovering over millions harboring latent infection, thus, calling for novel approaches for this dire ailment. In recent years, the non-calcemic physiological actions of vitamin D have drawn a great deal of attention. In this review, we will focus on the role of vitamin D in the innate immune defense against TB on the one hand and conversely on the immunomodulatory effects of vitamin D on autoimmunity. Taken together, the suggested dual role of vitamin D in treating TB infection and possibly preventing associated autoimmunity will constitute the basis of the current review. PMID- 19543862 TI - Is humeral segmental defect replacement device a stronger construct than locked IM nailing? AB - Intramedullary (IM) nailing is currently the most common method for treating patients with impending pathologic humeral fractures; however, this treatment is associated with known complications primarily owing to violation of the rotator cuff during insertion. A better option is needed. To determine if a humeral segmental replacement prosthesis would provide a stronger construct compared with an IM nail in this setting, we compared the mechanical properties of these two devices in a cadaver model simulating an impending pathologic fracture. In each of nine matched pairs of fresh human humeri one was randomly selected to undergo a 50% lateral middiaphyseal defect simulating an impending pathologic fracture and subsequent fixation with an IM nail and bone cement. The contralateral humerus underwent fixation using a humeral segmental defect prosthesis. We determined T-scores using DEXA. Each specimen subsequently was tested in torsion to failure. Peak torque and peak rotation at failure were greater for the prosthesis specimens whereas torsional stiffness was greater for the IM nail specimens. We found a linear relationship between peak torque and T-score for each device with the slopes of the lines suggesting the construct with the prosthesis can withstand greater forces than the IM nail and the differences between devices were greater in weaker bones. PMID- 19543863 TI - The John Charnley Award: The functional outcome of hip resurfacing and large-head THA is the same: a randomized, double-blind study. AB - Better functional outcome is believed by some to occur after hip resurfacing (HR) than conventional 28-mm total hip arthroplasty (THA) in young and active patients with hip osteoarthritis. However, the postulated superior outcome of HR over THA may simply be the result of a bias in patient selection or the use of a larger femoral head. We therefore asked whether HR would demonstrate superior functional outcome when compared with a THA with a large-diameter femoral head in a randomized, double-blind study. Gait speed and postural balance evaluations, functional tests, and clinical data were analyzed preoperatively and at 3, 6, and 12 months after surgery. Gait speed was used as the primary outcome measure. Forty-eight patients were randomized in the study and a third group of 14 healthy subjects served as controls. The gait speed and postural balance evaluations, the performance at most functional tests, and clinical scores were similar in HR and large-head THA groups at each followup period. The operated patients reached most control group values at 3 months postoperatively. By these measures, HR did not provide better clinical function over large-head THA. LEVEL OF EVIDENCE: Level I, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19543864 TI - Salvage of failed acetabular cages by nonbuttressed trabecular metal cups. AB - Acetabular revision of failed cages or rings may be facilitated by previously placed bone graft, enabling the use of highly porous cementless hemispheric cups. We retrospectively reviewed all 15 patients who had conversion of failed antiprotrusion cages (10 patients) or roof rings (five patients) to cementless cups. All patients had restoration of bone stock (three major column, eight morselized, four combined bone grafts) performed in conjunction with their index cage or ring reconstruction arthroplasty. The minimum followup was 24 months (average, 48.3 months; range, 24-72 months). Failure was defined as radiographic cup migration. In 12 of the 15 patients, there was no radiographic change in cup position at the last followup or symptoms indicative of loosening. The average Harris hip scores improved from 31 (range, 15-48) to 69 (range, 56-87) at latest followup. Cup failure occurred in three patients. In two patients, the failed cups were revised; the third patient refused additional surgery. Our experience suggests treatment of failed cages by highly porous cementless cups is a reasonable option. However, we recommend patients be followed closely to detect cup migration, which can occur until satisfactory bony ingrowth occurs. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19543867 TI - [Sexually transmitted diseases]. PMID- 19543865 TI - Outcomes after excision of pigmented villonodular synovitis of the knee. AB - Pigmented villonodular synovitis (PVNS) has a high but variable recurrence rate. Prior studies do not compare recurrence-free survival (RFS) for various surgical approaches or salvage surgery for relapse. We therefore determined: (1) RFS after excision; (2) RFS after salvage surgery for relapse; (3) factors associated with relapse. We retrospectively reviewed the medical records of 49 patients with previously untreated PVNS of the knee (12 localized, 37 diffuse) who were treated with synovectomy from 1991 to 2008; there were 22 males and 27 females, with mean age of 35.2 years (range, 10-73). Minimum followup was 1 year (mean, 6.2 years; range, 1-13). Twenty-one patients had a relapse. The RFS for index surgery was 75% and 53%; and for salvage surgery was 71% and 52% at 2 and 5 years respectively. The RFS was 95% for open versus 62% for arthroscopic synovectomy at 2 years, 71% and 41% at 5 years. The RFS was 91% for localized and 70% for diffuse PVNS at 2 years, 73% and 48% at 5 years. Diffuse disease (RR = 4.49) and arthroscopic synovectomy (RR = 3.30) were associated with relapse. Recurrence was frequent after synovectomy. Reexcision can salvage relapses as successfully as excision for primary disease; however, morbidity was associated with additional surgeries. LEVEL OF EVIDENCE: Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19543866 TI - [Prescription Tip: old and new]. PMID- 19543869 TI - [Activities of the Deutschen STD-Gesellschaft e.V. (Society for the Prevention of sexually transmitted diseases]]. PMID- 19543868 TI - ["Die grosse Barb" in the museum of the University of Marburg. An early documentation of acromegaly]. AB - The university museum for cultural history in the castle of Marburg has a portrait "Die grosse Barb", which represents a women suffering from acromegaly. She shows the typical pathologic alterations: thickening of the skin folds, thickening of the lips and the eyelids, growth of bones and cartilages, lengthening of the nose, enlargement of the ears, protrusion of the zygoma, mandible and the chin. Acromegaly is a consequence of enhanced secretion of growth hormone, which occurs also as a symptom of several syndromes, such as multiple endocrine neoplasia type 1, McCune-Albright-syndrome, and NAME syndrome (Carney complex type I). The most remarkable symptom of acromegaly is the gigantism. This occurs also in androgen-deficient states, such as the Klinefelter syndrome and some more genetic syndromes, of which the Simpson-Golabi-Behmel syndrome, the Sotos syndrome, the Marfan syndrome, the homocystinuria, and the fragile X-syndrome may be mentioned. Nothing is known on the further fate of the patient shown in the portrait. It is also unknown, whether she owes her position as a chambermaid to her gigantism, for it was a common use in courts to have people with abnormal body shapes in attendance. PMID- 19543870 TI - [Avoidance of wrong site surgery. Experiences by the introduction of measures for quality control and patient safety in a surgical casualty hospital]. AB - Wrong site surgery leads to substantial individual injury to the affected patient. Therefore, organizations such as the WHO, the Joint Commission International (JCI) and the Action Alliance Patient Safety (APS) have developed procedures for avoidance of wrong site surgery. This is based on the active incorporation of patients in the process of intervention identification. In this manner the disclosure to the patient, marking of the site of surgery, identification before induction of anesthesia and the team time out are documented in a checklist recommended by the APS before the operation begins. In the DRK Clinics in Berlin the procedure for avoidance of wrong site surgery was introduced before the second recertification by the JCI for implementation of the patient safety recommendations and compliance with the four stages documented in a standard form. For successful implementation all members of staff and in particular the heads of the clinics and departments must be actively involved in order to obtain the acceptance permanently and to ensure that wrong site surgery no longer occurs in the DRK Clinics Berlin. PMID- 19543871 TI - [Clean Hands Campaign. No chance for hospital infections!]. AB - There are approximately 500,000 hospital acquired infections per year in Germany of which about 20-30% (100,000-150,000) could be prevented. Hospital acquired infections are associated with increased mortality and prolonged hospital stay. Based on approximately 4 additional days of hospital stay, nosocomial infections cause additional 2 million hospital days per year. In other words, 6 hospitals each with 1,000 beds would be caring solely for patients with nosocomial infections for 1 year in Germany. Experts agree that careful hand hygiene is the single most effective measure to prevent transmission of pathogens. The rate of nosocomial infections can be reduced by improved compliance of hand hygiene as demonstrated in the literature. The ''AKTION Saubere Hande'' (Clean Hands Campaign) is a national campaign by the National Reference Centre for the Surveillance of Nosocomial Infections, the Society for Quality Management in the Health System and the Action Alliance Patient Safety aimed at the sustained improvement of hand hygiene behaviour in German hospitals. PMID- 19543872 TI - [Intracortical dedifferentiated chondrosarcoma]. AB - Chondrosarcoma is the second most frequent mesenchymal malignant tumour of the bone. Classification of this kind of tumour is made by clinical, radiological und pathological means. A case of an intracortical chondrosarcoma was first reported by Babinet et al. 2003 [2]. During the staging examination of a 59-year-old patient referred to our clinic because of a squamous cell carcinoma of the oropharynx, we also found a highly malignant intracortical dedifferentiated chondrosarcoma of the distal femur shaft. Due to the primary assumption of bone metastasis of the oropharynx tumour, marginal extralesional tumour resection was performed followed by composite osteosynthesis. Considerations on differential diagnosis and their implications for further therapy are discussed. PMID- 19543873 TI - [Standards of medical care for burn injuries, part 2]. PMID- 19543874 TI - [Plastic reconstructive surgery for burn injuries]. AB - The stage-adjusted therapy of thermal injuries is based on pathophysiologic mechanisms as well as functional and aesthetic requirements. Plastic reconstructive surgical approaches are highly important in the prevention of the frequent grave sequelae of thermal trauma and to achieve optimal functional rehabilitation and favourable outcome. In reconstructive surgery of burns operative goals are subdivided into acute, secondary reconstructive, functional and aesthetic indications. The achievement of early wound closure to preserve functional skin and soft tissue components is an essential part of acute reconstructive procedures. Functional reconstructive and aesthetic procedures supplement the conservative treatment modalities of the secondary phase of burn care with physical therapy, ergotherapy and psychological support. PMID- 19543875 TI - [Treatment of metacarpal fractures. Recommendations of the Hand Surgery Group of the German Trauma Society]. AB - The major goal in the treatment of metacarpal fractures is to restore the normal function of the hand. Radiological criteria and the clinical extent of displacement should be individually considered when taking the decision for or against conservative treatment. Internal fixation techniques must protect soft tissue structures. Small screws and plates have proven effective for head and shaft fractures, whereas intramedullary splinting is favoured for neck fractures. In instable and displaced fractures of the base of the first metacarpal, surgery is regularly performed to restore the bony shape and articular surface. To prevent functional impairments, early mobilization is desirable both during conservative treatment and following internal fixation. PMID- 19543877 TI - [Neuro-enhancement from an addiction specialist's viewpoint]. AB - The use of medications to enhance performance in healthy individuals is increasingly being propagated even by neuroscientists under such colorful terms as"neuro-enhancement". A large number of medications, including psychostimulants have been advocated in this context. Recent data from the German health insurance company DAK indicate that a number of employees already take medications to fulfill their professional needs and enhance performance. The use of new drugs in the healthy has even been advocated by prominent neurobiologists in a commentary in a recent edition of Nature. There are a number of ethical objections to this use and there is an apparent risk of addiction. To date German psychiatrists have surprisingly not been outspoken on this issue. The author would like to make an emphatic plea against"brain doping" in healthy individuals. PMID- 19543876 TI - Side biases in humans (Homo sapiens): three ecological studies on hemispheric asymmetries. AB - Hemispheric asymmetries and side biases have been studied in humans mostly in laboratory settings, and evidence obtained in naturalistic settings is scarce. We here report the results of three studies on human ear preference observed during social interactions in noisy environments, i.e., discotheques. In the first study, a spontaneous right-ear preference was observed during linguistic exchange between interacting individuals. This lateral bias was confirmed in a quasi experimental study in which a confederate experimenter evoked an ear-orienting response in bystanders, under the pretext of approaching them with a whispered request. In the last study, subjects showed a greater proneness to meet an experimenter's request when it was directly addressed to the right rather than the left ear. Our findings are in agreement both with laboratory studies on hemispheric lateralization for language and approach/avoidance behavior in humans and with animal research. The present work is one of the few studies demonstrating the natural expression of hemispheric asymmetries, showing their effect in everyday human behavior. PMID- 19543878 TI - [Urology update seminar UroUpdate. Dusseldorf 27th-28th February 2009]. PMID- 19543879 TI - [Immunomodulatory treatment approaches for prostate cancer]. AB - During the last 10 years different strategies for immunotherapy of prostate cancer have been investigated. These included unspecific and specific strategies to modulate or stimulate the immune system. For unspecific immunotherapy of prostate cancer innate humoral or cellular immune mechanisms are being stimulated, which are not specific to malignant cells. The global stimulation of the innate immune system is supposed to augment the immune reaction to prostate cancer by initiating an inflammatory reaction or other existing immune mechanisms. The main mediators and effectors of the unspecific immune system include humoral factors such as cytokines, complement system, and acute phase proteins and cellular components such as neutrophils, macrophages/monocytes, mast cells, and natural killer cells.In contrast, specific immunotherapy aims at adaptive immunity. This portion of the immune system can be amplified and thus specifically target tumor cells. Generation of a tumor-specific T cell reaction by vaccination or application of antibodies are the most promising approaches of specific immunotherapy. In a PubMed-based search of the current literature, publications regarding immunotherapy of prostate cancer were identified. The present article focuses on publications presenting clinical studies which investigate immunomodulatory treatments of prostate cancer. The results of these publications are described and discussed. PMID- 19543880 TI - Development and functional assessment of EST-derived 2RL-specific markers for 2BS.2RL translocations. AB - ESTs-derived markers are useful for comparative genomic analysis and can also serve as phenotype-linked functional markers. Here, we report the development of EST-derived 2RL-specific markers and the evaluation of the possibility of functional assessment of markers tagging 2RL, which carries Hessian fly resistance genes (loci). To identify transcripts specific to 2RL, unigene sequences in combination with wheat progenitor genomes were used. Total 275 contigs mapped to the long arms of homoeologous group 2 chromosomes were downloaded. To obtain a cluster corresponding to each of the wheat 275 contigs, unigene sequences of wheat, rice, barley, and rye were pooled for cross-species clusters. Out of 275 clusters examined, it was possible to design 112 cross species primer pairs for genome-specific amplifications. Out of 112 cross-species primer pairs, 45 primer pairs (40%) produced amplicons from at least one species (three wheat progenitors or rye). Among the 45 contigs, 73% were associated with one of known functions and 82% of the contigs associated with known functions were also associated with one of the GO categories. On the basis of the oligonucleotide sequence alignment of each of 45 genome-specific amplifications, 21 amplifications (47%) were suitable for designing RR genome-specific primers, which are specific to translocated rye chromatin 2RL. Six primer pairs (13%) successfully produced amplicons in the 2BS.2RL translocation lines and not in the non-2RLs. Functional assessment of one of the 2RL-specific markers, NSFT03P2_Contig4445, was performed on Hessian fly infested NILs. Under Hessian fly infestation, significantly high expression of a gene tagged by a 2RL-specific marker (NSFT03P2_Contig4445) was observed 1 day after infestation. EST-derived 2RL-specific marker development from this study provides a basis for the development of ESTs-derived markers for detecting wheat-rye translocations. In addition, these markers could be employed in elucidating functional analysis of genes on 2RL. PMID- 19543881 TI - Familial association between type 1 diabetes and other autoimmune and related diseases. AB - AIMS/HYPOTHESIS: In the era of genome-wide association studies, familial risks are used to estimate disease heritability and success in gene identification. We wanted to estimate associations between type 1 diabetes mellitus and 33 autoimmune and related diseases in parents, offspring, singleton siblings and twins. METHODS: The availability of a Multigeneration Register in Sweden provides reliable access to families throughout the last century. The diseases in individual family members were obtained through linkage to the Hospital Discharge Register. Standardised incidence ratios (SIRs) were calculated as relative risks of contracting type 1 diabetes in family members of affected patients compared with those lacking affected family members. RESULTS: Among a total of 450,899 patients, 21,168 were diagnosed with type 1 diabetes. Familial cases amounted to 10.3% of all type 1 diabetes patients. SIR for type 1 diabetes was 8.23 in offspring of affected parents, 11.92 in singleton siblings, 39.22 in multiplex families and 21.88 in twins; the calculated risk for monozygotic twins was 32.33. Type 1 diabetes in offspring was associated with 13 diseases in parents, including Addison's disease (SIR 2.41), asthma (1.38), coeliac disease (2.73), Graves' disease/hyperthyroidism (1.86), Hashimoto disease/hypothyroidism (2.35), pernicious anaemia (3.09), primary biliary cirrhosis (3.63), rheumatoid arthritis (2.12), sarcoidosis (1.62), systemic lupus erythematosus (2.04), ulcerative colitis (1.23) and Wegener's granulomatosis (2.12). CONCLUSIONS/INTERPRETATION: The concordant familial risks for type 1 diabetes were high and the calculated risk for multiplex families and monozygotic twins may be explained by epistatic gene x gene or gene x environment interactions. Familial associations with several autoimmune and related diseases suggest genetic sharing and challenge to gene identification. PMID- 19543882 TI - Is there an optimal breath pattern to minimize stress and strain during mechanical ventilation? AB - BACKGROUND: Inappropriate selection of tidal volume and rate on mechanical ventilators in patients with reduced lung volume may cause lung damage. In spite of this rather recent insight, the optimal breath pattern and the relative importance of elevating end-expiratory lung volume (EELV) are still debated. A recent hypothesis is that lung injury is caused by excessive stress and strain. This paper elaborates on that hypothesis and proposes a new approach to optimizing the breath pattern. METHODS: An index to quantify the impact of positive pressure ventilation on the lungs is defined (Stress-Strain Index, SSI) and calculated as a function of the breath pattern (tidal volume Vt and respiratory rate f) for five different levels of EELV. The breath pattern at which SSI is minimal (mSSI strategy) was compared with three other strategies: the "6 ml/kg Vt," minimal work of breathing and minimal force to breathe, for the different EELV levels. RESULTS: In the mathematical analysis, SSI was mainly determined by EELV and was much higher with low EELV. For each EELV level, a distinct minimum of SSI was found, defined by a particular Vt-f combination. The mSSI strategy yielded lower Vt and higher f (0.252 l and 39 b/min) as compared to the "6 ml/kg Vt" strategy (0.420 l and 17 b/min). CONCLUSION: The EELV is the main determinant of the SSI. For a given EELV, the SSI can be minimized by an optimal Vt-f combination. PMID- 19543884 TI - Determination of protein surface excess on a liquid/solid interface by single molecule counting. AB - Determination of protein surface excess is an important way of evaluating the properties of biomaterials and the characteristics of biosensors. A single molecule counting method is presented that uses a standard fluorescence microscope to measure coverage of a liquid/solid interface by adsorbed proteins. The extremely low surface excess of lysozyme and bovine serum albumin (BSA), in a bulk concentration range from 0.3 nmol L(-1) (0.02 microg mL(-1)) to 3 nmol L(-1) (0.2 microg mL(-1)), were measured by recording the counts of spatially isolated single molecules on either hydrophilic (glass) or hydrophobic (polydimethylsiloxane, PDMS) surfaces at different pH. The differences observed in amounts of adsorbed proteins under different experimental conditions can be qualitatively explained by the combined interactions of electrostatic and hydrophobic forces. This, in turn, implies that single-molecule counting is an effective way of measuring surface coverage at a liquid/solid interface. PMID- 19543885 TI - 3rd International IUPAC Symposium on Trace Elements in Food. PMID- 19543883 TI - Cognitive, psychomotor, and subjective effects of sodium oxybate and triazolam in healthy volunteers. AB - RATIONALE: Illicit gamma-hydroxybutyrate (GHB) has received attention as a "date rape drug" that produces robust amnesia; however, there is little experimental evidence in support of GHB's amnestic effects. OBJECTIVES: This study compared the cognitive effects of GHB (sodium oxybate) with those of triazolam in healthy volunteers. MATERIALS AND METHODS: Doses of sodium oxybate (1.125, 2.25, and 4.5 g/70 kg), triazolam (0.125, 0.25, and 0.5 mg/70 kg), and placebo were administered to 15 volunteers under repeated measures, counterbalanced, double blind, double-dummy conditions. The time course and peak physiological, psychomotor, subjective, and cognitive effects were examined. RESULTS: Sodium oxybate and triazolam produced similar increases in participant ratings of drug effects. Performance on psychomotor, working memory, and episodic memory tasks was impaired to a greater extent after triazolam than sodium oxybate. CONCLUSIONS: Together, these data suggest that sodium oxybate produces less psychomotor and cognitive impairment than triazolam at doses that produce equivalent participant-rated subjective effects in healthy volunteers. PMID- 19543886 TI - Forensic analysis of dyed textile fibers. AB - Textile fibers are a key form of trace evidence, and the ability to reliably associate or discriminate them is crucial for forensic scientists worldwide. While microscopic and instrumental analysis can be used to determine the composition of the fiber itself, additional specificity is gained by examining fiber color. This is particularly important when the bulk composition of the fiber is relatively uninformative, as it is with cotton, wool, or other natural fibers. Such analyses pose several problems, including extremely small sample sizes, the desire for nondestructive techniques, and the vast complexity of modern dye compositions. This review will focus on more recent methods for comparing fiber color by using chromatography, spectroscopy, and mass spectrometry. The increasing use of multivariate statistics and other data analysis techniques for the differentiation of spectra from dyed fibers will also be discussed. PMID- 19543888 TI - Incidental fat in the dural sinuses. PMID- 19543889 TI - Novel J stents reduce the risk of embolic stroke in vitro. AB - INTRODUCTION: Two and a half million Americans with atrial fibrillation are at an elevated risk for embolic stroke. Warfarin therapy is standard treatment for high risk patients, yet 40-65% of elderly patients do not receive anticoagulation therapy due to bleeding complications. To address this clinical need, we are evaluating a minimally invasive stent-based stroke prevention device to divert emboli from entering the arterial supply of the brain. METHODS: The feasibility of a J-shaped stroke prevention device was tested in a mock circulatory loop. Sixteen sets of 100 simulated emboli (1-5 mm(3)) were injected into the left atrium with and without J stents protecting the aortic arch vessels. To determine efficacy, emboli were trapped in filters in the aortic arch vessels and distal aorta for manual counting. RESULTS: J stents decreased the number of emboli that entered the brachiocephalic trunk by 93.7% (p < 0.0001), left common carotid artery by 79.8% (p < 0.0001), and left subclavian artery by 89.7% (p < 0.0001). CONCLUSIONS: In a mock circulation, J stents positioned in the aortic arch vessels and oriented downstream of aortic flow significantly decreased the number of emboli that entered the aortic arch vessels. These results warrant further investigation to determine the safety and efficacy of this prophylactic intervention to reduce embolic events, and chronic large animal studies are underway. PMID- 19543887 TI - Pharmacokinetics and dosage adjustment in patients with renal dysfunction. AB - INTRODUCTION: Chronic kidney disease is a common, progressive illness that is becoming a global public health problem. In patients with kidney dysfunction, the renal excretion of parent drug and/or its metabolites will be impaired, leading to their excessive accumulation in the body. In addition, the plasma protein binding of drugs may be significantly reduced, which in turn could influence the pharmacokinetic processes of distribution and elimination. The activity of several drug-metabolizing enzymes and drug transporters has been shown to be impaired in chronic renal failure. In patients with end-stage renal disease, dialysis techniques such as hemodialysis and continuous ambulatory peritoneal dialysis may remove drugs from the body, necessitating dosage adjustment. METHODS: Inappropriate dosing in patients with renal dysfunction can cause toxicity or ineffective therapy. Therefore, the normal dosage regimen of a drug may have to be adjusted in a patient with renal dysfunction. Dosage adjustment is based on the remaining kidney function, most often estimated on the basis of the patient's glomerular filtration rate (GFR) estimated by the Cockroft-Gault formula. Net renal excretion of drug is a combination of three processes: glomerular filtration, tubular secretion and tubular reabsorption. Therefore, dosage adjustment based on GFR may not always be appropriate and a re-evaluation of markers of renal function may be required. DISCUSSION: According to EMEA and FDA guidelines, a pharmacokinetic study should be carried out during the development phase of a new drug that is likely to be used in patients with renal dysfunction and whose pharmacokinetics are likely to be significantly altered in these patients. This study should be carried out in carefully selected subjects with varying degrees of renal dysfunction. In addition to this two-stage pharmacokinetic approach, a population PK/PD study in patients participating in phase II/phase III clinical trials can also be used to assess the impact of renal dysfunction on the drug's pharmacokinetics and pharmacodynamics. CONCLUSION: In conclusion, renal dysfunction affects more that just the renal handling of drugs and/or active drug metabolites. Even when the dosage adjustment recommended for patients with renal dysfunction are carefully followed, adverse drug reactions remain common. PMID- 19543891 TI - Effect of emodin on Candida albicans growth investigated by microcalorimetry combined with chemometric analysis. AB - Using the 3114/3115 thermal activity monitor (TAM) air isothermal microcalorimeter, ampoule mode, the heat output of Candida albicans growth at 37 degrees C was measured, and the effect of emodin on C. albicans growth was evaluated by microcalorimetry coupled with chemometric methods. The similarities between the heat flow power (HFP)-time curves of C. albicans growth affected by different concentrations of emodin were calculated by similarity analysis (SA). In the correspondence analysis (CA) diagram of eight quantitative parameters taken from the HFP-time curves, it could be deduced that emodin had definite dose effect relationship as the distance between different concentrations of it increased along with the dosage and the effect. From the principal component analysis (PCA) on eight quantitative parameters, the action of emodin on C. albicans growth could be easily evaluated by analyzing the change of values of the main two parameters, growth rate constant k (2) and maximum power output P(2)(m). The coherent results of SA, CA, and PCA showed that emodin at different concentrations had different effects on C. albicans growth metabolism: A low concentration (0-10 microg ml(-1)) poorly inhibited the growth of C. albicans, and a high concentration (15-35 microg ml(-1)) could notably inhibit growth of this fungus. This work provided a useful idea of the combination of microcalorimetry and chemometric analysis for investigating the effect of drug and other compounds on microbes. PMID- 19543892 TI - Cervical transforaminal epidural steroid injection for the management of cervical radiculopathy: a comparative study of particulate versus non-particulate steroids. AB - OBJECTIVE: To determine if a particulate steroid which has a risk for embolic infarct would be more effective than a non-particulate steroid for transforaminal epidural steroid injection (TFESI). The purpose of this study was (1) to compare the effect of cervical TFESI using particulate (e.g., triamcinolone) and non particulate (e.g., dexamethasone) steroids and (2) to evaluate the effectiveness of cervical TFESI in general. MATERIALS AND METHODS: From January 2006 to August 2008, 159 consecutive patients [male:female (M:F) 89:70; mean age 53 years, range 33-75 years] who underwent cervical TFESI were included in this non-randomized study. For cervical TFESI, triamcinolone was injected into 97 patients and dexamethasone into 62 patients. Short-term follow-up was conducted within 1 month. The outcome was classified as effective or ineffective. Fisher's exact test was used to analyze the difference of outcome according to the injected steroid (triamcinolone vs dexamethasone). Other possible outcome predictors, such as age, gender, duration of radiculopathy, predominant symptom, attack of radiculopathy, cause of radiculopathy, number of nerve root compression levels, previous operation, and failure of previous interlaminar epidural injection, were also analyzed. RESULTS: Cervical TFESI using triamcinolone (78/97, 80.4%) was slightly more effective than that using dexamethasone (43/62, 69.4%), which was not significant (P = 0.129). In general, cervical TFESIs were effective in 121 of 159 patients (76.1%) at short-term follow-up. The only significant outcome predictor was whether the patient had had a previous operation (6/13, 46/2%) or not (115/146, 78.8%) (P = 0.015). CONCLUSION: There was no significant difference between particulate or non-particulate steroid for the effect of cervical TFESI. Cervical TFESI was effective in managing cervical radiculopathy in general. PMID- 19543893 TI - Supraspinatus tendon tears: comparison of 3D US and MR arthrography with surgical correlation. AB - OBJECTIVE: The objective of the study was to compare the diagnostic reliability of 3D US with MR arthrography in diagnosing supraspinatus tendon tears, with arthroscopic findings used as the standard. MATERIALS AND METHODS: In a prospective study 50 patients who later underwent arthroscopic surgery of the rotator cuff were examined pre-operatively by 3D US with MR arthrography. The presence or absence of a full- or partial-thickness supraspinatus tendon tear and the tear size as demonstrated by each imaging and arthroscopy was recorded. The tear size was divided into three grades: small (<1 cm), medium (1-3 cm), and large (>3 cm). RESULTS: The arthroscopic diagnosis was a full-thickness tear in 40 patients, partial-thickness tears in 5, and intact supraspinatus tendon in 5. 3D US correctly diagnosed 35 out of 40 full-thickness tears and MR arthrography 39 out of 40 full-thickness tears. Regarding partial-thickness tears, 3D US underestimated 2 cases as no tear and overestimated 1 case as a full-thickness tear. MR arthrography underestimated 1 case as a partial-thickness tear and overestimated 2 cases as full-thickness and partial-thickness tears respectively. 3D US and MR arthrography yield a sensitivity for full-thickness tears of 87.5% and 97.5% with specificity of 90.0% and 90.0%. Based on the grading system, 3D US measurements correctly predicted the tear size of 23 (65.7%) of the 35 full thickness tears and MR arthrography 30 (75.0%) of the 39 full-thickness tears. CONCLUSION: Three-dimensional ultrasound seems to be a promising imaging modality comparable to MR arthrography for the assessment of the supraspinatus tendon tears. PMID- 19543894 TI - Jaccoud's arthropathy and psoriatic arthritis, a rare association. AB - Jaccoud's arthropathy is a syndrome of chronic progressive painless deformity of the hands and feet with surprisingly well-preserved functions. Although it is most frequently seen in patients with systemic lupus erythematosus, an association with other diseases has occasionally been described. A patient with long-standing psoriasis arthritis is presented in this report who developed an arthropathy with the clinical and radiologic finding indistinguishable from that of the Jaccoud's arthropathy. In patients with long-standing psoriasis arthritis, the disease may be associated with Jaccoud's arthropathy. PMID- 19543895 TI - Ultrasonography shows disappearance of monosodium urate crystal deposition on hyaline cartilage after sustained normouricemia is achieved. AB - This study aimed at determining whether lowering serum urate (SU) to less than 6 mg/dl in patients with gout affects ultrasonographic findings. Seven joints in five patients with monosodium urate (MSU) crystal proven gout and hyperuricemia were examined over time with serial ultrasonography. Four of the five patients were treated with urate lowering drugs (ULDs) (allopurinol, n = 3; probenecid, n = 1). One patient was treated with colchicine alone. Attention was given to changes in a hyperechoic, irregular coating of the hyaline cartilage in the examined joints (double contour sign or "urate icing"). This coating was considered to represent precipitate of MSU crystals. Index joints included metacarpophalangeal (MCP) joints (n = 2), knee joints (n = 3), and first metatarsophalangeal (MTP) joints (n = 2). The interval between baseline and follow-up images ranged from 7 to 18 months. Serial SU levels were obtained during the follow-up period. During the follow-up period, three patients treated with ULD (allopurinol, n = 2; probenecid, n = 1) achieved a SU level of <6 mg/dl. In two patients, SU levels remained above 6 mg/dl (treated with allopurinol, n = 1; treated with colchicine, n = 1). At baseline, the double contour sign was seen in all patients. In those patients who achieved SU levels of <6 ml/dl, this sign had disappeared at follow-up. Disappearance of the double contour sign was seen in two knee joints, two first MTP joints, and one MCP joint. In contrast, disappearance of the double contour sign was not seen in patients who maintained a SU level > or =7 mg/dl. In one patient treated with allopurinol, SU levels improved from 13 to 7 mg/dl during the follow-up period. Decrease, but not resolution of the hyperechoic coating was seen in this patient. In the patient treated with colchicine alone, SU levels remained >8 mg/dl, and no sonographic change was observed. In our patients, sonographic signs of deposition of MSU crystals on the surface of hyaline cartilage disappeared completely if sustained normouricemia was achieved. This is the first report showing that characteristic sonographic changes are influenced by ULDs once SU levels remain < or =6 mg/dl for 7 months or more. Sonographic changes of gout correlate with SU levels and may be a non-invasive means to track changes in the uric acid pool. Larger prospective studies are needed to further assess these potentially important findings. PMID- 19543896 TI - Twitchin of mollusc smooth muscles can induce "catch"-like properties in human skeletal muscle: support for the assumption that the "catch" state involves twitchin linkages between myofilaments. AB - Molluscan catch muscles can maintain tension with low or even no energy utilization, and therefore, they represent ideal models for studying energy saving holding states. For many decades it was assumed that catch is due to a simple slowing of the force-generating myosin head cross-bridge cycles. However, recently evidences increased suggesting that catch is rather caused by passive structures linking the myofilaments in a phosphorylation-dependent manner. One possible linkage structure is the titin-like thick filament protein twitchin, which could form bridges to the thin filaments. Twitchin is known to regulate the catch state depending on its phosphorylation state. Here, we found that twitchin induces a catch-like stiffness in skinned human skeletal muscle fibres, when these fibres are exposed to this protein. Subsequent phosphorylation of twitchin reduces the stiffness. These findings support the assumption that catch of molluscan smooth muscle involves twitchin linkages between thick and thin filaments. PMID- 19543898 TI - Colon replacement of vagina to restore menstrual function in 11 adolescent girls with vaginal or cervicovaginal agenesis. AB - AIM: Cervicovaginal or vaginal agenesis with functioning endometrial tissue is rare. We report the construction of a colon conduit which is anastomosed to posterior uterine wall or upper vaginal pouch to allow menstruation. MATERIALS AND METHODS: We report seven girls with cervicovaginal agenesis and four with lower vaginal agenesis (aged 12-20 years) who presented with painful cryptomenorrheoa. All the girls wanted to conserve their uterus and menstruate normally. A colon conduit was constructed for the egress of menstrual blood. The colon conduit was anastomosed to the posterior uterine wall in the seven girls with cervicovaginal agenesis and to the distended upper vaginal pouch in the four girls with vaginal agenesis. Utero-colonic neovaginal anastomosis was performed only after excising a circular portion of the posterior myometrium to prevent stenosis. RESULTS: The colon conduit functioned effectively, providing an egress for regular painless menstruation. One patient had stenosis of the perineal neovaginal orifice for which dilations were done. One girl has married and reports satisfactory intercourse. The mean follow up is 2.2 years. CONCLUSIONS: This group of patients forms a separate subgroup needing a conduit not only for sexual function but also for menstruation. However, if treated by the method described herein, they should be cautioned against pregnancy if they have cervicovaginal agenesis and against vaginal delivery if they have vaginal agenesis. PMID- 19543897 TI - Melatonin attenuates the acetylcholine-induced contraction in isolated intestine of a teleost fish. AB - The present study investigates the possible direct actions of melatonin (N-acetyl 5-methoxytryptamine) on intestinal motility in goldfish (Carassius auratus) using an in vitro system of isolated intestine in an organ bath engaged to an isometric transducer. The longitudinal strips from goldfish intestine in the organ bath showed a resting spontaneous myogenic rhythmic activity which is not altered by melatonin. The addition of acetylcholine (1 nmol l(-1)-10 mmol l(-1)) to the organ bath induces a significant contraction of the intestinal strips in a concentration-dependent manner. The addition of melatonin and its agonist, 2 iodomelatonin, induced a concentration-dependent attenuation of acetylcholine induced contractile response. The specificity of this effect is tested by the preincubation of the intestine strips in the presence of two melatoninergic antagonists, luzindole (a non-selective MT(1)/MT(2) melatonin receptor antagonist) and 4-P-PDOT (preferred antagonist of MT2 receptor subtype), which counteracted the melatonin-induced relaxation in a concentration-dependent manner. Finally, present results demonstrate that this melatoninergic effect on intestinal strips is a process highly dependent on extracellular calcium. In conclusion, this is the first study demonstrating the role of melatonin in the control of gut motility in a non-mammalian vertebrate. The melatonin effects on isolated intestine from goldfish are mediated by melatoninergic membrane receptors, and could suggest a delay in food transit time, supporting its anorectic effect reported on in vivo studies. PMID- 19543900 TI - Are cosmetic products which include an SPF appropriate for daily use? AB - The goal of this study is to investigate commercially available cosmetics (foundations, skin care creams) which also claim to include a sun protection factor (SPF). Are these products, which are not considered sunscreen products, helpful or could they be harmful? Using an in vitro method, we tested the effectiveness of 35 commercially available products against UVB and UVA radiation. For each product, our testing focused on determining the following four values in terms of current legal recommendations: SPF, UVA protection factor (PF-UVA), UVB/UVA ratio and critical wavelength (lambda(c)). We also tested each product's level of photostability. Effectively, when considering instructions for use (skincare products are applied once, in the morning) any product displaying an SPF must be particularly photostable, since its labeling does not indicate reapplication. In contrast, the packaging on sunscreen products clearly indicates the necessity of frequent reapplication. Out of the 35 products we tested, seven do not comply with legislation regarding sunscreen products. This non-compliance translates into insufficient protection against UVA radiation. The products sold in pharmacies did comply. In terms of photostability, only eight products out of the original 35 proved to be sufficiently photostable. It would seem inappropriate to use filters in the formulas of non-sunscreen cosmetics. PMID- 19543901 TI - A novel point mutation at donor splice-site in intron 42 of type III collagen gene resulting in the inclusion of 30 nucleotides into the mature mRNA in a case of vascular type of Ehlers-Danlos syndrome. AB - Vascular type of Ehlers-Danlos syndrome (EDS) is the most severe type of EDS. It is an autosomal dominantly inherited disorder that results from mutations within the alpha1 type III collagen gene (COL3A1). We report a novel point mutation at donor splice-site in intron 42 of type III collagen gene resulting in the inclusion of 30 nucleotides into the mature mRNA in a case of vascular type of EDS. Since the age of approximately 8 months, the patient had had repeated episodes of purpura and gradually developed thin, translucent skin. She had a past history of pneumothorax. At the initial examination, she was found to have the characteristic facies, i.e., bird-like face, of the vascular type of EDS, thinning of skin over the limbs and trunk, and scattered purpura. The blood vessels under the skin could be clearly visualized. She showed hypermobility of the small joints of all the four limbs and acrogeric changes of the hands and feet. Analysis of the amount of collagen synthesized from cultured dermal fibroblasts by SDS-polyacrylamide gel electrophoresis and fluorography was conducted based on the clinical suspicion of the vascular type of EDS, and a marked reduction in the synthesis of type III collagen was observed. Genetic analysis of the COL3A1 revealed a novel point mutation at the donor splice-site of intron 42, which resulted in the inclusion of 30 nucleotides into the mature mRNA of one allele. PMID- 19543899 TI - Analysis of mutations in TP53, APC, K-ras, and DCC genes in the non-dysplastic mucosa of patients with inflammatory bowel disease. AB - BACKGROUND AND AIMS: Patients with ulcerative colitis (UC) and Crohn's disease (CD) have a high risk for colorectal cancer (CRC). To understand the molecular basis of colitis-associated CRC, we analyzed alterations in TP53, APC, K-ras, and DCC genes in the non-dysplastic UC and CD colon. MATERIALS AND METHODS: Endoscopic biopsies were collected from six predefined colon sites of 35 UC and 12 CD patients for DNA extraction and genetic analysis. RESULTS: A mutation was found in codon 1141 of the APC gene of two CD patients, being somatic in one and germinative in the other. The mutation seen in both patients was a base exchange of thymine for cytosine, resulting in an exchange of leucine for serine. We did not detect any mutations in the other samples analyzed. CONCLUSIONS: Mutations in APC gene may occur in the non-dysplastic CD mucosa of patients with disease for more than 10 years. The follow-up of these patients will show the likelihood of mutant APC progressing to CRC in CD. Further analysis will be required for evaluating the impact of these findings in the context of cancer surveillance in inflammatory bowel disease. PMID- 19543902 TI - Impaired cutaneous wound healing with excess granulation tissue formation in TNFalpha-null mice. AB - We examined the effects of lacking tumor necrosis factor alpha (TNFalpha) on the healing process of a cutaneous wound in mice using TNFalpha-deficient mice. A full-thickness circular cutaneous wound 5.0 mm in diameter was produced in the dorsal skin of wild-type (WT) or TNFalpha-null (KO) mice. After specific intervals of healing, the healing pattern was evaluated by macroscopic observation, histology, immunohistochemistry, or real-time reverse transcription polymerase chain reaction. Effect of Smad7 gene transfer on the healing phenotype of KO mice was also examined. The results showed that loss of TNFalpha promotes granulation tissue formation and retards reepithelialization in a circular wound in mouse dorsal skin. Immunohistochemistry showed that distribution of macrophages and myofibroblasts in newly generated granulation tissue seemed similar between WT and KO mice. However, lacking TNFalpha enhanced mRNA expression of TGFbeta1 and collagen Ialpha2 in such tissue. Smad7 gene transfer counteracted excess granulation tissue formation in KO mice. In conclusion, lacking TNFalpha potentiates Smad-mediated fibrogenic reaction in healing dermis and retards reepithelialization in a healing mouse cutaneous wound. PMID- 19543903 TI - Posterior colpotomy revisited: a forgotten route for retrieving larger benign ovarian lesions following laparoscopic excision. AB - INTRODUCTION: Although simple, ovarian cysts can be decompressed (within an impermeable pouch) following laparoscopic excision to allow lesions of up to 10 cm to be delivered safely. It is more difficult to remove solid or semi-solid ovarian tumours (such as fibromas and dermoid cysts) through this route. We present a modification of laparoscopic-assisted transvaginal retrieval of ovarian tumours through a posterior colpotomy incision (in a bag) and suggest that this is a route that allows large specimens to be retrieved safely and with minimal spillage. METHOD: Women with solid or semi-solid ovarian cysts of greater than 5 cm, undergoing operative laparoscopy, were offered the option of having the specimens delivered through a posterior colpotomy, if retrieval through the abdominal ports proved difficult. RESULTS: The median specimen diameter was 6 cm (range 4-8 cm) and the median blood loss was 200 ml (range 150-250 ml). The median operating time was 50 min (range 40-90 min) with most patients being discharged within 24 h (median 18 h; range 16-21 h). There was no inadvertent spillage of cyst content or any intra or post-operative complications. CONCLUSION: Laparoscopic-assisted transvaginal removal of these lesions (in a bag!) allows large specimens to be removed securely and with minimal spillage. PMID- 19543904 TI - 15-Year review of laryngeal and oral dysplasias and progression to invasive carcinoma. AB - The objective of this study is to identify a management or follow-up strategy for patients with laryngeal and oral dysplasia. A chart review of all patients with laryngeal and oral dysplasia over a 15-year period was performed. All patients were followed for a minimum period of 5 years from initial diagnosis of oropharyngeal or laryngeal dysplasia. If invasive carcinoma was demonstrated on subsequent biopsies, the exact time of this was recorded and Kaplan-Meier survival curves were plotted. In the laryngeal cohort, 45 patients were identified, 15 (33%) developed carcinoma, 7 of 30 patients (23%) with mild or moderate dysplasia, compared with 8 of 15 (53%) with severe dysplasia or CIS (P = 0.01). Thirteen of the carcinomas (87%) developed within 36 months of original biopsy. In the oral cohort, 32 patients were identified, 17 (53%) developed carcinoma, 1 of 9 patients (11%) with mild dysplasia, compared with 8 of 12 (67%) with severe dysplasia and 8 of 10 (80%) with CIS (P < 0.001). Fifteen of the 17 patients (88%) developed carcinoma within 36 months of original biopsy. In conclusion, although numbers are small, our results show that mild and moderate laryngeal dysplasia behaves differently to severe dysplasia and CIS. Mild oral dysplasia also behaves differently to severe dysplasia or CIS. In general, progress to malignancy happens within a 3-year period. Severe dysplasia or CIS should be managed aggressively. PMID- 19543905 TI - Population stratification in Argentina strongly influences likelihood ratio estimates in paternity testing as revealed by a simulation-based approach. AB - A simulation-based analysis was carried out to investigate the potential effects of population substructure in paternity testing in Argentina. The study was performed by evaluating paternity indexes (PI) calculated from different simulated pedigree scenarios and using 15 autosomal short tandem repeats (STRs) from eight Argentinean databases. The results show important statistically significant differences between PI values depending on the dataset employed. These differences are more dramatic when considering Native American versus urban populations. This study also indicates that the use of Fst to correct for the effect of population stratification on PI might be inappropriate because it cannot account for the particularities of single paternity cases. PMID- 19543907 TI - The acute effects of running on blood pressure estimation using pulse transit time in normotensive subjects. AB - Pulse transit time (PTT) is a potential parameter for cuffless blood pressure (BP) estimation. Since exercise induces changes in arterial properties that can influence the relationship between BP and PTT, we investigate whether PTT can be used to estimate BP after successive bouts of exercise. PTT-foot, PTT-peak (time intervals from the peak of electrocardiogram R-wave to the foot and peak of photoplethysmogram, respectively) and BP of 41 normotensive subjects (aged 25 +/- 4 years) were measured in the first test. A repeatability test was then conducted on 14 subjects after 6 months. Each test included two periods of running on the treadmill at 10 and 8 km/h (with a rest in between). In both tests, systolic BP (SBP) was closely correlated with PTT-foot and PTT-peak. For each subject, the best fit linear relationships between SBP and PTTs were determined over all phases of each test. The differences between the linear fits and measured data were greater after the second period of running for all subjects in both tests. This implied that the relationships started to change after the second period of running. When SBP in the repeatability test was predicted using the linear regression coefficients from the first test, the linear fit after the first period of exercise was still better than after the second. The repeated observations in both tests suggest that PTT is a potential parameter for cuffless BP estimation after one period of exercise, but would need re-calibration (relationship between BP and PTTs) for measurements after successive phases of exercise. PMID- 19543906 TI - Visualizing vitreous in vitrectomy by triamcinolone. AB - BACKGROUND: Visualizing vitreous and retinal surface during vitrectomy, choromovitrectomy, is a novel approach in vitrectomy. METHODS: Triamcinolone acetonide (TA) is now most commonly used as an adjunct to vitrectomy for this purpose. Intraoperative use of TA can visualize posterior hyaloid, preretinal membrane, internal limiting membrane (ILM) during pars plana vitrectomy, and prolapsed vitreous during cataract surgery, which can make the procedure very safe and effective. TA-assisted vitrectomy is favorably applied to surgery for macular hole, proliferative vitreoretinopathy, diabetic retinopathy, uveitis, and others. In addition, this technique can disclose the residual hyaloid cortex pattern after surgical posterior vitreous detachment. Diffuse posterior hyaloids cortex is frequently seen in diabetic retinopathy and high myopia, and an island like cortex is often left on the macula, which can be a scaffold of future macular pucker. RESULTS: The prospective controlled clinical trial showed that TA assisted vitrectomy reduced the incidence of intraoperative retinal break and retinal detachment more significantly than conventional vitrectomy, although post operative visual acuity after 1 year was almost the same with each method. Adverse events related to TA-assisted vitrectomy included transient intraocular pressure elevation (approximately 5.0%); most of these events are manageable with topical treatment. The incidence of acute endophthalmitis was 0.03-0.05% , which was at a level comparable to conventional vitrectomy. CONCLUSIONS: In this article, current status and the possible problems of TA-assisted vitrectomy are reviewed. PMID- 19543908 TI - Different response to eccentric and concentric training in older men and women. AB - Sarcopenia is the age-related loss of muscle mass and strength and has been associated with an increased risk of falling and the development of metabolic diseases. Various training protocols, nutritional and hormonal interventions have been proposed to prevent sarcopenia. This study explores the potential of continuous eccentric exercise to retard age-related loss of muscle mass and function. Elderly men and women (80.6 +/- 3.5 years) were randomized to one of three training interventions demanding a training effort of two sessions weekly for 12 weeks: cognitive training (CT; n = 16), conventional resistance training (RET; n = 23) and eccentric ergometer training (EET; n = 23). Subjects were tested for functional parameters and body composition. Biopsies were collected from M. vastus lateralis before and after the intervention for the assessment of fiber size and composition. Maximal isometric leg extension strength (MEL: +8.4 +/- 1.7%) and eccentric muscle coordination (COORD: -43 +/- 4%) were significantly improved with EET but not with RET (MEL: +2.3 +/- 2.0%; COORD: -13 +/- 3%) and CT (MEL: -2.3 +/- 2.5%; COORD: -12 +/- 5%), respectively. We observed a loss of body fat (-5.0 +/- 1.1%) and thigh fat (-6.9 +/- 1.5%) in EET subjects only. Relative thigh lean mass increased with EET (+2.5 +/- 0.6%) and RET (+2.0 +/- 0.3%) and correlated negatively with type IIX/type II muscle fiber ratios. It was concluded that both RET and EET are beneficial for the elderly with regard to muscle functional and structural improvements but differ in their spectrum of effects. A training frequency of only two sessions per week seems to be the lower limit for a training stimulus to reveal measurable benefits. PMID- 19543909 TI - One night of sleep deprivation decreases treadmill endurance performance. AB - The aim was to test the hypothesis that one night of sleep deprivation will impair pre-loaded 30 min endurance performance and alter the cardio-respiratory, thermoregulatory and perceptual responses to exercise. Eleven males completed two randomised trials separated by 7 days: once after normal sleep (496 (18) min: CON) and once following 30 h without sleep (SDEP). After 30 h participants performed a 30 min pre-load at 60% [VO(2 max) followed by a 30 min self-paced treadmill distance test. Speed, RPE, core temperature (T(re)), mean skin temperature (T(sk)), heart rate (HR) and respiratory parameters VO(2 max), VCO(2), VE, RER pre-load only) were measured. Less distance (P = 0.016, d = 0.23) was covered in the distance test after SDEP (6037 (759) 95%CI 5527 to 6547 m) compared with CON (6224 (818) 95%CI 5674 to 6773 m). SDEP did not significantly alter T(re) at rest or thermoregulatory responses during the pre-load including heat storage (0.8 degrees C) and T(sk). With the exception of raised VO(2) at 30 min on the pre-load, cardio-respiratory parameters, RPE and speed were not different between trials during the pre-load or distance test (distance test mean HR, CON 174 (12), SDEP 170 (13) beats min(-1): mean RPE, CON 14.8 (2.7), SDEP 14.9 (2.6)). In conclusion, one night of sleep deprivation decreased endurance performance with limited effect on pacing, cardio-respiratory or thermoregulatory function. Despite running less distance after sleep deprivation compared with control, participants' perception of effort was similar indicating that altered perception of effort may account for decreased endurance performance after a night without sleep. PMID- 19543910 TI - Physiological responses to simulated stair climbing in professional firefighters wearing rubber and leather boots. AB - No studies have considered whether a firefighter's boots are a factor influencing physiological responses. The purpose of this study was to examine physiological responses to a fire simulation activity (stair climb) in professional firefighters wearing rubber boots (RB) and leather boots (LB). Twelve professional firefighters participated in two counterbalanced simulated firefighter stair climb (SFSC) sessions, one wearing RB and the other wearing LB. Heart rate, oxygen uptake (VO(2)), expiratory ventilation (V(E)), blood lactate (BLa), salivary cortisol (SCORT), and leg strength were assessed prior to and following a SFSC. LB elicited significantly greater SCORT values and knee flexion time to peak torque. Furthermore, RB revealed significantly greater ankle dorsiflexion peak torque after SFSC. BLa was positively related to knee flexion peak torque after SFSC in the RB. Firefighters when wearing the RB may be more effective at resisting fatigue and increase more force production. PMID- 19543911 TI - Development of the rhopalial nervous system in Aurelia sp.1 (Cnidaria, Scyphozoa). AB - We examined the development of the nervous system in the rhopalium, a medusa specific sensory structure, in Aurelia sp.1 (Cnidaria, Scyphozoa) using confocal microscopy. The rhopalial nervous system appears primarily ectodermal and contains neurons immunoreactive to antibodies against tyrosinated tubulin, taurine, GLWamide, and FMRFamide. The rhopalial nervous system develops in an ordered manner: the presumptive gravity-sensing organ, consisting of the lithocyst and the touch plate, differentiates first; the "marginal center," which controls swimming activity, second; and finally, the ocelli, the presumptive photoreceptors. At least seven bilaterally arranged neuronal clusters consisting of sensory and ganglion cells and their neuronal processes became evident in the rhopalium during metamorphosis to the medusa stage. Our analysis provides an anatomical framework for future gene expression and experimental studies of development and functions of scyphozoan rhopalia. PMID- 19543912 TI - Thyroid tumours of uncertain malignant potential: frequency and diagnostic reproducibility. AB - The term thyroid tumours of uncertain malignant potential (TT-UMP) has been proposed for a subgroup of follicular-patterned thyroid tumours for which benignancy or malignancy cannot be assessed with certainty. The frequency, diagnostic reproducibility, immunohistochemistry and molecular genetic profiling of such tumours have been poorly explored. We, therefore, investigated (1) the frequency of TT-UMP diagnosed in a single institution (Nice, France: 2004-2008), (2) the observer variation among four pathologists, (3) whether immunohistochemical and molecular genetic profiling of TT-UMP provide additional information concerning such lesions. A series of 31 diagnosed TT-UMP (2.9%) out of 1,078 consecutive thyroidectomies were analysed. It comprised 15 follicular thyroid tumours of UMP (FT-UMP) and 16 well-differentiated tumours of UMP (WDT UMP). Observer concordance was 70% for all TT-UMP. More than 50% of FT-UMP expressed galectin-3 and CK19, whereas more than 50% of WDT-UMP expressed HBME-1. Five cases of TT-UMP showed N-RAS mutations, while one showed H-RAS mutation and another PAX8/PPARgamma rearrangement. In conclusion, the frequency of TT-UMP is low in our institution. Diagnostic reproducibility is within the same range as other published data on follicular-patterned thyroid tumours. The ancillary methods have a low impact on aiding diagnosis of such lesions. PMID- 19543913 TI - Novel swine-origin influenza A virus in humans: another pandemic knocking at the door. AB - Influenza A viruses represent a continuous pandemic threat. In April 2009, a novel influenza A virus, the so-called swine-origin influenza A (H1N1) virus (S OIV), was identified in Mexico. Although S-OIV originates from triple-reassortant swine influenza A (H1) that has been circulating in North American pig herds since the end of the 1990s, S-OIV is readily transmitted between humans but is not epidemic in pigs. After its discovery, S-OIV rapidly spread throughout the world within few weeks. In this review, we sum up the current situation and put it into the context of the current state of knowledge of influenza and influenza pandemics. Some indications suggest that a pandemic may be mild but even "mild" pandemics can result in millions of deaths. However, no reasonable forecasts how this pandemic may develop can be made at this time. Despite stockpiling by many countries and WHO, antiviral drugs will be limited in case of pandemic and resistances may emerge. Effective vaccines are regarded to be crucial for the control of influenza pandemics. However, production capacities are restricted and development/production of a S-OIV vaccine will interfere with manufacturing of seasonal influenza vaccines. The authors are convinced that S-OIV should be taken seriously as pandemic threat and underestimation of the menace by S-OIV to be by far more dangerous than its overestimation. PMID- 19543914 TI - Overweight is associated with improved cancer-specific survival in patients with organ-confined renal cell carcinoma. AB - OBJECTIVES: Overweight/obesity is known to increase the risk of developing renal cell carcinoma (RCC). However, data on the prognostic impact of overweight in RCC is still conflicting. We assessed whether different body mass index (BMI) levels at the time of surgery had an effect on the long-term prognosis of RCC patients. METHODS: We evaluated 771 patients, with complete information about their BMI, who had undergone renal surgery for RCC between 1990 and 2005 at the authors' institution; the mean follow-up was 5.48 years. RESULTS: Underweight, normal weight, pre-obesity, and obesity were diagnosed in 4 (0.5%), 239 (31%), 356 (46.2%), and 172 (22.3%) RCC patients, respectively. Overweight (BMI >25) was significantly associated with younger age (P = 0.004) and positive nodal status (P = 0.04) but not with tumor stage, grade, visceral metastasis, gender, histological subtype, or tumor-related symptoms. Overweight patients had a significantly lower risk of cancer-related death; their median 5-year tumor specific survival rate was 80% as opposed to 72% for patients with a BMI below 25 (P = 0.003). Interestingly, subgroup analysis revealed that the positive association between overweight and survival was even more pronounced in organ confined (P < 0.001) RCC, but no correlation was observed in advanced disease (P = 0.23). CONCLUSION: We were able to identify overweight as an independent prognostic marker of improved tumor-specific survival in patients with organ confined RCC. Basic research is required to resolve the dilemma of why, if a higher BMI predisposes to RCC, it concurrently prolongs survival after patients have undergone (partial) nephrectomy. PMID- 19543916 TI - Light and electron microscopic studies on the development of the ovaries of Culex pipiens quinquefasciatus (Say) (Diptera: Culicidae). AB - Christopher's stages of ovarian development are redefined in Culex pipiens quinquefasciatus to provide a more useful description of mosquito physiology. During this ovarian development, seven growing stages of the oocyte are observed in the adult stage. Two distinct periods of egg follicle growth, the previtellogenesis and vitellogenesis, are seen first. The gradual increase in the oocyte size during these phases is attributed to yolk deposition. During vitellogenesis, the oocyte and nurse cell nucleus are similar in size (stage I). Then, the oocyte enlarges and occupies one third of the egg follicle, while the nurse cell (NC) occupies the remaining two thirds. At this phase, the yolk granules (stage III) enlarge, occupying about half of the egg follicle, while the NC contain the other half (stage IV). An increase and accumulation of yolk granules leads to the filling of about three fourths of the egg bulk and makes the oocyte nucleus hardly visible during stage V. The oocyte undergoes a remarkable growth, occupying nine tenth of the length of the follicle in stage VI. Stage VII corresponds to a mature cigar-shaped egg and contains two kinds of yolk globules: a large one and a small one. The oocyte and the NC at all stages are surrounded by a single layer of epithelial cells which increase in number and form the vitelline membrane and the chorion. The mitochondria in the NC, oocyte, and follicular epithelium appear in light microscopy as granules of different sizes. The Golgi apparatus appears in different sizes and shapes, since in the early stages of development, it is restricted to the perinuclear zone. When the development proceeds, it enlarges in size, spreads over the whole cytoplasm, and participates in the formation of the yolk. PMID- 19543915 TI - Plasmodium falciparum and Plasmodium vivax: so similar, yet very different. AB - The recently published whole genome sequence information of one of the human malaria parasites, Plasmodium vivax, have provided opportunities to compare similar features with Plasmodium falciparum that causes the most deadly form of human malaria. We herewith present comparative genomic insights into the whole genome of the two parasites and also to several other characteristics in terms of disease pathogenecity, evolution, etc. We show that while high similarities exist at the functional gene level, several contrasting features for other characteristics are hallmarks of these two human malaria parasites. PMID- 19543917 TI - Functional response of staging semipalmated sandpipers feeding on burrowing amphipods. AB - Despite its fundamental relevance to many ecological processes in predator-prey relationships, the functional response, which relates predator intake rate to prey density, remains difficult to document in the field. Here, I document the functional response of semipalmated sandpipers (Calidris pusilla) foraging on a burrowing amphipod Corophium volutator during three field seasons at the peak of fall migration in the upper Bay of Fundy (New Brunswick, Canada). I gathered data during the ebbing tide when all sandpipers are highly motivated to feed after a lengthy hide-tide fast. As birds follow the receding tideline, foragers encounter prey at different densities and do not aggregate in the richest food patches. Results show that intake rate increased at a decreasing rate with Corophium density, yielding a type II functional response typical of many shorebird species. Intake rate decreased in the later stages of migration stopover at a time where preferred prey items have been shown to occur at lower densities due to prior depletion. At this period of lower prey availability, intake rate also decreased with sandpiper density providing evidence for interference at low prey density. The results illustrate the fact that the functional response may not be unique but instead vary as a function of the type of competitive relationship among foragers. PMID- 19543918 TI - Life history plasticity and fitness in a caddisfly in response to proximate cues of pond-drying. AB - Pond-drying is a model for understanding the causes of life history variation in metamorphic organisms. However, we know relatively little about how interactions among specific proximate cues of pond-drying affect juvenile life history, how those responses might be mitigated by diet, and the post-metamorphic consequences for adult fitness. I manipulated larval diet, water depth, and water temperature during the aquatic larval stage of a temporary pond-dwelling caddisfly, Limnephilus indivisus. I predicted that shallow depths and warm temperatures (depth x temperature) associated with pond-drying would have negative effects on larval survival, growth, development, adult size, female fecundity, and adult longevity, but that supplementation of the larval diet should mitigate the trade off between juvenile growth and pre-reproductive mortality risk by ameliorating the negative effects of pond-drying (diet x depth, diet x temperature) on these traits. Larval survival was enhanced by diet supplementation but was not affected by depth or temperature. Larval diet and water temperatures acted independently on growth, development, and female size, and growth rates were higher when larval diets were supplemented relative to ambient diets; development times were shorter when temperatures were warmer relative to colder; adult females were larger when larvae were fed a supplemented diet but smaller when reared in warm water. Larval growth and development were not affected by depth, but female size was reduced under shallow relative to deep conditions. Female longevity and fecundity were affected by the larval diet x female size interaction. Surprisingly, this was independent of the depth x temperature interaction on female longevity and fecundity suggesting that reductions in adult fitness due to juvenile abiotic conditions can be independent of size-at-maturity. Future studies should quantify the effect of proximate cues of pond-drying on juvenile survival and life history as well as adult fitness correlates. PMID- 19543919 TI - The role of trematode parasites in larval anuran communities: an aquatic ecologist's guide to the major players. AB - Conservation strategies depend on our understanding of the ecosystem and community dynamics. To date, such understanding has focused mostly on predator prey and competitor interactions. It is increasingly clear, however, that parasite-host interactions may represent a large, and important, component of natural communities. The need to consider multiple factors and their synergistic interactions if we are to elucidate the contribution of anthropogenic factors to loss in biodiversity is exemplified by research into present-day amphibian declines. Only recently has the role of factors such as trematode parasite infections been incorporated into studies of the population and community dynamics of aquatic systems. We argue that this is due, at least in part, to difficulties faced by aquatic ecologists in sifting through the complex systematics that pervade the parasite literature. We note that two trematode species are of dominant importance with regard to North American larval anuran communities, and provide in this review a clear explanation of how to distinguish between the infective stages of these two parasites. We describe the general biology and life history of these parasites, as well as what is known about their effect on larval anurans, and the interactive effects of environmental stressors (typically anthropogenic in nature) and parasites on larval anurans. We hope that this review will convince the reader of the potential importance of these parasites to aquatic communities in general, and to amphibian communities specifically, and will also provide the information necessary for aquatic ecologists to more frequently consider the role of these parasites in their studies of aquatic ecology. PMID- 19543920 TI - Nonlinear effects of winter sea ice on the survival probabilities of Adelie penguins. AB - The population dynamics of Antarctic seabirds are influenced by variations in winter sea ice extent and persistence; however, the type of relationship differs according to the region and the demographic parameter considered. We used annual presence/absence data obtained from 1,138 individually marked birds to study the influence of environmental and individual characteristics on the survival of Adelie penguins Pygoscelis adeliae at Edmonson Point (Ross Sea, Antarctica) between 1994 and 2005. About 25% of 600 birds marked as chicks were reobserved at the natal colony. The capture and survival rates of Adelie penguins at this colony increased with the age of individuals, and five age classes were identified for both parameters. Mean adult survival was 0.85 (SE = 0.01), and no effect of sex on survival was evident. Breeding propensity, as measured by adult capture rates, was close to one, indicating a constant breeding effort through time. Temporal variations in survival were best explained by a quadratic relationship with winter sea ice extent anomalies in the Ross Sea, suggesting that for this region optimal conditions are intermediate between too much and too little winter sea ice. This is likely the result of a balance between suitable wintering habitat and food availability. Survival rates were not correlated with the Southern Oscillation Index. Low adult survival after a season characterized by severe environmental conditions at breeding but favorable conditions during winter suggested an additional mortality mediated by the reproductive effort. Adelie penguins are sensitive indicators of environmental changes in the Antarctic, and the results from this study provide insights into regional responses of this species to variability in winter sea ice habitat. PMID- 19543921 TI - Thermal radiation absorbed by dairy cows in pasture. AB - The goal of the present paper was to assess a method for estimating the thermal radiation absorbed by dairy cows (0.875 Holstein-0.125 Guzerath) on pasture. A field test was conducted with 472 crossbred dairy cows in three locations of a tropical region. The following environmental data were collected: air temperature, partial vapour pressure, wind speed, black globe temperature, ground surface temperature and solar radiation. Average total radiation absorbed by animals was calculated as R(abs) = 640.0 +/- 3.1 W .m(-2). Absorbed short-wave radiation (solar direct, diffuse and reflected) averaged 297.9 +/- 2.7 W m(-2); long wave (from the sky and from terrestrial surfaces) averaged 342.1 +/- 1.5 W m(-2). It was suggested that a new environmental measurement, the effective radiant heat load (ERHL), could be used to assess the effective mean radiant temperature (T*(mr)). Average T*(mr) was 101.4 +/- 1.2 degrees C, in contrast to the usual mean radiant temperature, T(mr) = 65.1 +/- 0.5 degrees C. Estimates of T*(mr) were considered as more reliable than those of T (mr) in evaluating the thermal environment in the open field, because T (mr) is almost totally associated only with long wave radiation. PMID- 19543922 TI - Chemistry, antiproliferative properties, tumor selectivity, and molecular mechanisms of novel gold(III) compounds for cancer treatment: a systematic study. AB - The antiproliferative properties of a group of 13 structurally diverse gold(III) compounds, including six mononuclear gold(III) complexes, five dinuclear oxo bridged gold(III) complexes, and two organogold(III) compounds, toward several human tumor cell lines were evaluated in vitro using a systematic screening strategy. Initially all compounds were tested against a panel of 12 human tumor cell lines, and the best performers were tested against a larger 36-cell-line panel. Very pronounced antiproliferative properties were highlighted in most cases, with cytotoxic potencies commonly falling in the low micromolar--and even nanomolar--range. Overall, good-to-excellent tumor selectivity was established for at least seven compounds, making them particularly attractive for further pharmacological evaluation. Compare analysis suggested that the observed antiproliferative effects are caused by a variety of molecular mechanisms, in most cases "DNA-independent," and completely different from those of platinum drugs. Remarkably, some new biomolecular systems such as histone deacetylase, protein kinase C/staurosporine, mammalian target of rapamycin/rapamycin, and cyclin-dependent kinases were proposed for the first time as likely biochemical targets for the gold(III) species investigated. The results conclusively qualify gold(III) compounds as a promising class of cytotoxic agents, of outstanding interest for cancer treatment, while providing initial insight into their modes of action. PMID- 19543923 TI - Molecular dynamics simulations of mouse ferrochelatase variants: what distorts and orientates the porphyrin? AB - Molecular dynamics simulations of the wild-type and variant forms of the mouse ferrochelatase in complex with the product (haem) have been performed using the GROMOS96 force field, in the NpT ensemble. Ferrochelatase, the last enzyme in the catalytic pathway of the haem biosynthesis, catalyses the reaction of insertion of a ferrous ion into protoporphyrin IX by distorting the planar geometry of the latter reactant. The simulations presented aim at understanding the role of active-site residues in this catalytic process. Analysis of the simulation trajectories explains the consequences of the mutations introduced and sheds more light on the role of the His209 residue in porphyrin macrocycle distortion. The function of residues coordinating propionate groups of the haem molecule is discussed in terms of stability of the substrate and product complexes. PMID- 19543924 TI - Interaction between cyanobacterial copper chaperone Atx1 and zinc homeostasis. AB - Cyanobacterial Atx1 is a copper chaperone which interacts with two copper transporting ATPases to assist copper supply to plastocyanin and cytochrome oxidase. ZiaA is a Zn(2+)-exporting ATPase and ziaA expression is regulated by ZiaR. Here we show that gene expression from the ziaA operator promoter, monitored using reverse transcriptase PCR and lacZ fusions, is elevated in Deltaatx1 mutants. Although Cu(+) tightly binds recombinant ZiaR in vitro, Cu(+) is less effective at dissociating ZiaR-DNA complexes than Zn(2+) and crucially ziaA expression responds to Zn(2+) but not copper in both wild-type and Deltaatx1 cells. Consistent with enhanced expression of ZiaA, Deltaatx1 cells have slightly elevated Zn(2+) resistance. Recombinant Zn(2+)-Atx1 is recovered from Zn(2+) supplemented Escherichia coli and even after copper supplementation substantial amounts of Zn(2+)-Atx1 are isolated. Taken together, these data suggest that Zn(2+)-Atx1 can form in vivo. PMID- 19543925 TI - The effect of thread design on stress distribution in a solid screw implant: a 3D finite element analysis. AB - The biomechanical behavior of implant thread plays an important role on stresses at implant-bone interface. Information about the effect of different thread profiles upon the bone stresses is limited. The purpose of this study was to evaluate the effects of different implant thread designs on stress distribution characteristics at supporting structures. In this study, three-dimensional (3D) finite element (FE) stress-analysis method was used. Four types of 3D mathematical models simulating four different thread-form configurations for a solid screw implant was prepared with supporting bone structure. V-thread (1), buttress (2), reverse buttress (3), and square thread designs were simulated. A 100-N static axial occlusal load was applied to occlusal surface of abutment to calculate the stress distributions. Solidworks/Cosmosworks structural analysis programs were used for FE modeling/analysis. The analysis of the von Mises stress values revealed that maximum stress concentrations were located at loading areas of implant abutments and cervical cortical bone regions for all models. Stress concentration at cortical bone (18.3 MPa) was higher than spongious bone (13.3 MPa), and concentration of first thread (18 MPa) was higher than other threads (13.3 MPa). It was seen that, while the von Mises stress distribution patterns at different implant thread models were similar, the concentration of compressive stresses were different. The present study showed that the use of different thread form designs did not affect the von Mises concentration at supporting bone structure. However, the compressive stress concentrations differ by various thread profiles. PMID- 19543927 TI - An early oral health care program starting during pregnancy: results of a prospective clinical long-term study. AB - This study covers phase IV of a prospective clinical long-term study. Objective of this clinical investigation was to analyze the effects of a long-term prevention program on dental and oral health of teenagers at the age of 13 to 14 years. The entire study was subdivided into four phases. Phase I comprised an individual preventive care during pregnancy ("primary-primary prevention"); phase II assessed mothers and their young children until the age of 3 years ("primary prevention"); and in phase III, mothers and children at the age of 6 years were investigated. In phase IV of the study, the oral health of 13- to 14-year-old teenagers was examined (13.4 +/- 0.5 years; n = 29). All phases consisted of an examination, education about oral health care, and treatment based on the concept of an early oral health care promotion. The control group consisted of randomly selected adolescents at the same age (n = 30). The following clinical parameters were assessed: decayed/missing/filled teeth (DMF-T)/decayed, missing, and filled surface teeth index, hygiene index, papilla bleeding index, Periodontal Screening Index, and Streptococcus mutans/Lactobacillus concentration in saliva. The teenagers of the "prevention" group of phase IV of our prospective study revealed a share of 89.7% caries-free dentitions (65.5% sound; 24.2% caries-free with fillings). Mean DMF-T was 0.55 +/- 1.0. The control group showed a significantly higher mean DMF-T of 1.5 +/- 1.5 (p < 0.05) and revealed 56.7% of caries-free dentitions (30% sound, 26.7% caries-free with restorations). Our data clearly document that an early oral health care promotion starting during pregnancy may cause a sustained and long-term improvement of the oral health of children. PMID- 19543926 TI - Undesirable and adverse effects of tooth-whitening products: a review. AB - Hydrogen peroxide (H(2)O(2)) is a powerful oxidising agent. It gives rise to agents known to be effective bleaching agents. The mechanisms of bleaching involve the degradation of the extracellular matrix and oxidation of chromophores located within enamel and dentin. However, H(2)O(2) produces also local undesirable effects on tooth structures and oral mucosa. In clinical conditions, the daily low-level doses used to produce tooth whitening never generate general acute and sub-acute toxic effects. Genotoxicity and carcinogenicity only occur at concentrations that are never reached during dental treatments. Some transient adverse effects have been reported on the oral mucosa and the digestive tract if the product is swallowed. Local effects may occur on the oral mucosa and dental tissues during whitening, namely, pulp sensitivity, cervical resorption, release of selected components of dental restorative materials, and alteration of the enamel surface. Most of the local effects are dependent of the technique and concentration of the product so far used, but as the results of bleaching obtained are not stable, repeated treatments add to the adverse effects. The informed decision to administer or not and the control of bleaching effects should stand in the hand of dental surgeons and certainly not as it appears at present, as cosmetics sold without any restriction despite the potential health hazards of peroxides. PMID- 19543930 TI - New jet-aeration system using 'Supercavitation'. AB - BACKGROUND, AIM AND SCOPE: A newly developed fine bubble aeration system, by which air is transferred under supercavitation conditions, shows a clearly better performance than traditional, well-known aerators that rely on the jet-pump principle and its performance can be compared to oxygen transfer rates achieved in membrane and foil plate aerators. MATERIALS AND METHOD: A prototype supercavitation aerator installed at a sewage treatment plant revealed an air input rate, which was about one third lower than that of the jet-pump system, which it replaced. RESULTS: In spite of this low air input rate, the daily demand of pure oxygen for the additionally installed membrane aeration system went down by approximately 49%, from the original level of about 1,200 m(3)/day to about 600 m(3)/day-and this over a test period of more than 7 months. CONCLUSIONS AND DISCUSSION: The observed high oxygen transfer rates cannot be explained by traditional mass transfer mechanisms. It is assumed that a large amount of water being transferred into the gas phase by supercavitation contacting directly oxygen also in the gas phase and thereby overcoming mass transfer hindrances which might be favoured by hydroxyl radicals. With this new aerator, during the first 3 months of test phase, already more than 10,000 Euros had been saved because of the reduced pure oxygen demand. PMID- 19543928 TI - Molecular-docking-guided 3D-QSAR studies of substituted isoquinoline-1,3-(2H,4H) diones as cyclin-dependent kinase 4 (CDK4) inhibitors. AB - The cyclin-dependent kinases (CDKs) are critical regulators of cell cycle progression, and are involved in uncontrolled cell proliferation-a hallmark of cancer. This suggests that small molecular inhibitors of CDKs might be attractive as prospective antitumor agents. To explore the relationship between the structures of substituted isoquinoline-1,3-(2H,4H)-diones and their inhibition of CDK4, 3D-QSAR studies were performed on a dataset of 48 compounds. The bioactive conformation of template compound 34 was obtained by performing molecular docking into the ATP binding site of the homology model of CDK4 and ranking by highest consensus score, which was then used to build and align the rest of the molecules in the series. The constructed comparative molecular similarity indices analysis (CoMSIA) produces significantly better results than comparative molecular field analysis (CoMFA), with r(2)(cv) = 0.707 and r(2) = 0.988. The contours analysis provides useful information about the structural requirements for substituted isoquinoline-1,3-(2H,4H)-diones for CDK4 inhibitory activity. PMID- 19543929 TI - Ammonium acetate enhances solvent production by Clostridium acetobutylicum EA 2018 using cassava as a fermentation medium. AB - Cassava, due to its high starch content and low cost, is a promising candidate substrate for large-scale fermentation processes aimed at producing the solvents acetone, butanol and ethanol (ABE). However, the solvent yield from the fermentation of cassava reaches only 60% of that achieved by fermenting corn. We have found that the addition of ammonium acetate (CH(3)COONH(4)) to the cassava medium significantly promotes solvent production from cassava fermented by Clostridium acetobutylicum EA 2018, a mutant with a high butanol ratio. When cassava medium was supplemented with 30 mM ammonium acetate, the acetone, butanol and total solvent production reached 5.0, 13.0 and 19.4 g/l, respectively, after 48 h of fermentation. This level of solvent production is comparable to that obtained from corn medium. Both ammonium (NH(4) (+)) and acetate (CH(3)COO(-)) were required for increased solvent synthesis. We also demonstrated substantially increased acetic and butyric acid accumulation during the acidogenesis phase as well as greater acid re-assimilation during the solventogenesis period in ammonium acetate-supplemented cassava medium. Reverse transcription-polymerase chain reaction analysis indicated that the transcription of several genes encoding enzymes related to acidogenesis and solventogenesis in C. acetobutylicum EA 2018 were enhanced by the addition of ammonium acetate to the cassava medium. PMID- 19543931 TI - The optimal linear transformation-based fMRI feature space analysis. AB - This paper proposes a method of extending the optimal linear transformation (OLT), an image analysis technique of feature space, from magnetic resonance imaging (MRI) to functional magnetic resonance imaging (fMRI) so as to improve the activation detection performance over conventional approaches of fMRI analysis. The method was: (1) ideal hemodynamic responses for different stimuli were generated by convolving the theoretical hemodynamic response model with the stimulus timing, (2) considering the ideal hemodynamic responses as hypothetical signature vectors for different activity patterns of interest, OLT was used to extract the features of fMRI data. The resultant feature space had particular geometric clustering properties. It was then classified into different groups, each pertaining to an activity pattern of interest; the applied signature vector for each group was obtained by averaging, (3) using the applied signature vectors, OLT was applied again to generate fMRI composite images with high SNRs for the desired activity patterns. Simulations and a blocked fMRI experiment were employed to validate the proposed method. The simulation and the experiment results indicated the proposed method was capable of improving some conventional methods to be more sensitive to activations, having strong contrast between activations and inactivations, and being more valid for complex activity patterns. PMID- 19543932 TI - Metabolites from an endophytic fungus sphaceloma sp. LN-15 isolated from the leaves of Melia azedarach. AB - Two new natural compounds, a symmetrical disulfide dimer didodecyl 3,3'' dithiodipropionate (1) and a pregnane steroid 5,16-pregnadien-3beta-ol-20-one acetate (2), were isolated together with two known compounds, ergosta 4,6,8(14),22-tetraen-3-one (3) and ergosterol peroxide (4), from the ethyl acetate soluble extract of fermentation broth of an endophytic fungus, Sphaceloma sp. LN-15 isolated from the leaves of Melia azedarach L. and grown in pure culture. Their structures were determined on the basis of spectroscopic methods including 1D and 2D nuclear magnetic resonance spectroscopy (NMR) experiments and by mass spectrometric measurements (MS). These fungal metabolites were isolated for the first time from the genus Sphaceloma. The structure of 1 was also confirmed by chemical synthesis. PMID- 19543933 TI - [Pulmonary vein isolation with multipolar ablation catheters for the treatment of paroxysmal atrial fibrillation]. AB - Atrial fibrillation (AFIB) is the most common atrial rhythm disease and is associated with an increased risk of thromboembolic and cardiac complications. Different therapies are used in clinical routine: frequency control with anticoagulants and rhythm control. In patients with paroxysmal AFIB but without structural heart disease, antiarrhythmic drug therapy was previously first priority; however, pulmonary vein catheter ablation is becoming more important. "Single tip" systems, ultrasound, and various balloon techniques have been used in clinical routine. New radiofrequency systems with multipolar radiofrequency ablation catheters, however, give hope for better outcomes with fewer intraprocedural complications, as well as shorter procedure and fluoroscopy times. PMID- 19543935 TI - Felix Vicq d'Azyr (1748-1794). PMID- 19543936 TI - Maternal representations, infant psychiatric status, and mother-child relationship in clinic-referred and non-referred infants. AB - To examine the relations between maternal representations, infant socio-emotional difficulties, and mother-child relational behavior, 49 clinic-referred infants and their mothers were compared to 30 non-referred controls. Clinic-referred infants' psychiatric status was determined with the DC 0-3-R classification of Zeanah and Benoit (Child Adolesc Psychiatry Clin N Am 4:539-554, 1995) and controls were screened for socio-emotional difficulties. Mothers were interviewed with the parent development interview (Aber et al. in The parent development interview. Unpublished manuscript, 1985) and dyads were observed in free play and problem-solving interactions. Group differences emerged for maternal representations and relational behaviors. Representations of clinic-referred mothers were characterized by lower joy, coherence, and richness, and higher anger experienced in the mother-infant relationship compared to controls. During free play, clinic-referred mothers showed lower sensitivity and higher intrusiveness and provided less adequate instrumental and emotional assistance and support during problem solving. Referred children showed lower social engagement during free play. Associations were found among maternal representations, maternal interactive behavior, child social engagement, and the child's ability to self-regulate during a challenging task. These findings provide empirical support for theoretical and clinical perspectives suggesting a reciprocal link between maternal negative representations and mother and child's maladaptive behaviors in the context of early socio-emotional difficulties and mental health referrals. PMID- 19543937 TI - Bacterial community and nitrogen fixation in the red turpentine beetle, Dendroctonus valens LeConte (Coleoptera: Curculionidae: Scolytinae). AB - The red turpentine beetle, Dendroctonus valens LeConte (Coleoptera: Curculionidae: Scolytinae), colonizes all pines species within its native range throughout North and Central America. Recently, this species was accidentally introduced to China, where it has caused severe damage in pine forests. It belongs to a group of beetles that spend most of their lives between the tree bark and sapwood, where it feeds on phloem: a poor substrate with very low nutritional value of nitrogen and toxic properties due to its high content of secondary defensive compounds. The aim of this study was to characterize the bacterial community of the D. valens gut by culture-dependent and -independent methods. Polymerase chain reaction denaturing gradient gel electrophoresis and ribosomal gene library analyses revealed that species diversity in the D. valens gut was relatively low, containing between six and 17 bacterial species. The bacterial community associated with larvae and adults was dominated by members of the following genera: Lactococcus, Acinetobacter, Pantoea, Rahnella, Stenothrophomonas, Erwinia, Enterobacter, Serratia, Janibacter, Leifsonia, Cellulomonas, and Cellulosimicrobium. The members of the last four genera showed cellulolytic activity in vitro and could be involved in cellulose breakdown in the insect gut. Finally, nitrogen fixation was demonstrated in live larvae and adults; however, capacity of nitrogen fixing in vitro was not found among enterobacterial species isolated in nitrogen-free media; neither were nifD nor nifH genes detected. In contrast, nifD gen was detected in metagenomic DNA from insect guts. The identification of bacterial species and their potential physiological capacities will allow exploring the role of gut symbiotic bacteria in the adaptation and survival of D. valens in a harsh chemical habitat poor in nitrogen sources. PMID- 19543938 TI - Benthic ecology of Vibrio spp. and pathogenic Vibrio species in a coastal Mediterranean environment (La Spezia Gulf, Italy). AB - We carried out a 16-month in situ study to investigate the ecology of Vibrio spp. and pathogenic Vibrio species in coastal sediments of the Mediterranean Sea, employing multiple-regression analysis to reveal the major environmental factors controlling their occurrence in the benthic environment. In addition, association between vibrios and sediment-inhabiting meiofauna, which is a major component of benthic ecosystems, was investigated. Culturable and total Vibrio spp. estimates by most-probable-number technique coupled with standard polymerase chain reaction (PCR) and real-time PCR methods, respectively, were at least one order of magnitude higher in sediment than in seawater. In addition, potential human pathogenic species Vibrio cholerae, Vibrio vulnificus and Vibrio parahaemolyticus occurred in the sediment with V. parahaemolyticus being the most frequently found. In the pelagic environment, 60% of total variance in culturable Vibrio data was explained by sea surface temperature (40%), salinity (13%) and organic matter concentration (7%). In the benthic environment, sea surface temperature was the only factor that significantly affected culturable Vibrio occurrence although it explained only 25% of total variance, suggesting that additional unexplored factors may play a role as well. No correlation was found between culturable Vibrio spp. concentrations and the abundance of harpacticoid copepods in the sediment whilst a negative correlation was found between Vibrio spp. and nematode abundance which accounted for almost 90% of the total meiofaunal density. Taxonomic analysis revealed that selective bacterial feeders accounted for nearly 50% of the total nematode community and included genera such as Terschellingia, Molgolaimus and Halalaimus, suggesting that top-down control by nematode grazing may be an important factor affecting Vibrio occurrence in these sediments. It is concluded that the benthic marine environment may function as a reservoir of Vibrio spp. and potential pathogenic vibrios whose ecological features appeared substantially different from the ones recognised in the pelagic environment. PMID- 19543939 TI - A double-blind, randomized, placebo-controlled, two-dose comparative study of botulinum toxin type A for treating glabellar lines in Japanese subjects: what if sample size and statistical tests mattered? PMID- 19543941 TI - Crocodile attacks in Australia: challenges for injury prevention and trauma care. AB - INTRODUCTION: Saltwater crocodiles are formidable predators in northern Australia, and crocodile attacks on humans are not rare. With recent deaths highlighting this as a public health issue, an evidence-based discourse about effective methods of minimizing the danger to humans is needed. METHODS: Using the Haddon Matrix for injury prevention, approaches to minimizing crocodile associated death and injury were sought. RESULTS: Possibilities for harm minimization before, during and after a crocodile attack are identified, and their merits appraised. The importance of excellent prehospital and surgical and critical care is emphasized. CONCLUSIONS: A combination of behavior adaptation, mutual respect, and minimizing contact will be the key to minimizing the harm from attacks, and excellent medical and surgical care will always be necessary for those unfortunate to be victims but fortunate to survive. PMID- 19543940 TI - One-stage augmentation-mastopexy with Wise pattern and inverted-T scar. PMID- 19543942 TI - Long-term results of percutaneous bilioenteric anastomotic stricture treatment in liver-transplanted children. AB - The purpose of this study was to evaluate the mid- and long-term results of percutaneous transhepatic cholangiography (PTC) and biliary drainage in children with isolated bilioenteric anastomotic stenosis (BAS) after pediatric liver transplantation. Sixty-four children underwent PTC from March 1993 to May 2008. Nineteen cholangiograms were normal; 10 showed intrahepatic biliary stenosis and BAS, and 35 showed isolated BAS. Cadaveric grafts were used in 19 and living donor grafts in 16 patients. Four patients received a whole liver, and 31 patients received a left lobe or left lateral segment. Roux-en-Y hepaticojejunostomy was performed in all patients. Indication for PTC was based on clinical, laboratory, and histopathologic findings. In patients with isolated BAS, dilation and biliary catheter placement, with changes every 2 months, were performed. Patients were separated into 4 groups according to number of treatment sessions required. The drainage catheter was removed if cholangiogram showed no significant residual stenosis and normal biliary emptying time after a minimum of 6 months. The relationship between risk factors (recipient's weight <10 kg, previous exposure to Cytomegalovirus, donor-recipient sex and weight relations, autoimmune disease as indication for transplantion, previous Kasai's surgery, use of reduced liver grafts, chronic or acute rejection occurrence) and treatment was evaluated. Before PTC, fever was observed in 46%, biliary dilation in 23%, increased bilirubin in 57%, and increased gamma-glutamyltransferase (GGT) in 100% of patients. In the group with BAS, 24 of 35 (69%) patients had histopathologic findings of cholestasis as did 9 of 19 (47%) patients in the group with normal PTC. Of the 35 patients, 23 (65.7%) needed 1 (group I), 7 needed 2 (group II), 4 needed 3 (group III), and 1 needed 4 treatment sessions (group IV). The best results were observed after 1 treatment session, and the mean duration of catheter placement and replacement was 10 months. The primary patency rate was 61.2%, and the recurrence rate was 34.3% (group I). Seven patients (7 of 35; 20%) had their stricture treated with a second treatment session (group II). The average drainage time in group II was 24 months. During a period >20 months, 4 patients (4 of 35; 11.4%) required 1 additional treatment session (group III), and 1 patient (1 of 35; 2.9%) had a catheter placed at the end of the study period (group IV). Drainage time in group I was significantly shorter than those in groups II, III, and IV (p < 0.05). There was no statistically significant relation between therapeutic response and the selected risk factors (p > 0.05). The majority of complications, such as catheter displacement and leakage, were classified as minor; however, 2 patients (5.7%) with hemobilia were noted. Complications increased according to the need for reintervention. In conclusion, balloon dilation and percutaneous drainage placement is safe and effective, and it has long-term patency for children with BAS after liver transplantation. Because of prolonged treatment time, reintervention may be necessary, thereby increasing the complication rate. Balloon dilation and percutaneous drainage placement should be considered as the first treatment option because of its minimally invasive nature. PMID- 19543943 TI - Archaeal diversity in deep-sea sediments estimated by means of different terminal restriction fragment length polymorphisms (T-RFLP) protocols. AB - Despite the increasing recognition of the quantitative importance of Archaea in all marine systems, the protocols for a rapid estimate of Archaeal diversity patterns in deep-sea sediments have been only poorly tested yet. Sediment samples from 11 deep-sea sites (from 79 degrees N to 36 degrees N, at depths comprised from 469 to 5,571 m) were used to compare the performance of two different primer sets (ARCH21f/ARCH958r and ARCH109f/ARCH 915r) and three restriction enzymes (AluI, Rsa I, and HaeIII) for the fingerprinting analysis of Archaeal diversity using terminal-restriction fragment length polymorphisms (T-RFLP). In silico and experimental analyses indicated that different combinations of primer sets and restriction enzymes provided different values of benthic Archaeal ribotype richness and different Archaeal assemblage compositions. The use of the ARCH109f/ARCH 915r primer set in combination with AluI provided the best results (a number of ribotypes up to four folds higher than other combinations), suggesting that this primer set should be used in future studies dealing with the analysis of the patterns of Archaeal diversity in deep-sea sediments. Multivariate multiple regression analysis revealed that, whatever the T-RFLP protocol utilized, latitude and temperature explained most of the variance in benthic Archaeal ribotype richness, while water depth had a negligible role. PMID- 19543945 TI - Biodegradation of crude oil by an Arctic psychrotrophic bacterium Pseudoalteromomas sp. P29. AB - A psychrotrophic petroleum-degrading bacterium Pseudoalteromonas sp. P29 was isolated from marine sediment, which was collected during 2nd Chinese Arctic Scientific Expedition. The phenotypic character and biodegradation efficiency on mixed oil or vacuum oil were tested at low temperature. The strain Pseudoalteromonas sp. P29 grew in a range of temperature from 5 to 35 degrees C and the optimum temperature was 25 degrees C. Gas chromatography analysis indicated that the strain might preferentially metabolize shorter-chain alkanes. The biodegradation efficiency were nearly 90 and 80%, respectively, after incubation at 5 degrees C for 28 days in the mineral medium supplement with mixed oil or vacuum oil as the sole carbon and energy source. The results showed a possible exploitation of the strain in future biotechnological processes especially in cold contaminated environments. PMID- 19543944 TI - Enhanced degradation of TNT by genome-shuffled Stenotrophomonas maltophilia OK-5. AB - In this study, the enhanced degradation of TNT using cultures of genome-shuffled Stenotrophomonas maltophilia OK-5 mt-3 has been examined and the proteome of shuffled strain was compared to the wild-type OK-5 strain. Genome shuffling of S. maltophilia OK-5 was used to achieve a rapid enhancement of TNT degradation. The initial mutant population was generated by NTG treatment and UV irradiation. The wild-type OK-5 strain was able to degrade 0.2 mM TNT within 6 days, yet barely tolerated 0.5 mM TNT while the shuffled OK-5 mt-3 was capable of completely degrading 0.5 mM TNT within 8 days, and 1.2 mM within 24 days. The proteomic analysis of the shuffled OK-5 mt-3 demonstrated the changes in the expression levels of certain proteins compared to wild-type OK-5. These results provide clues for understanding TNT tolerance and improved TNT degradation by shuffled S. maltophilia OK-5 mt-3 and have possible applications in the processing of industrial waste containing relatively high TNT concentrations. PMID- 19543946 TI - 131I-Tositumomab (Bexxar) vs. 90Y-Ibritumomab (Zevalin) therapy of low-grade refractory/relapsed non-Hodgkin lymphoma. AB - INTRODUCTION: The American Cancer Society estimated 66,120 new cases of non Hodgkin lymphoma (NHL) in the USA in 2008. Radioimmunotherapy has been shown in clinical trials to be an effective treatment for refractory/relapsed NHL. The available agents are Bexxar, a (131)I radiolabeled murine monoclonal antibody, and Zevalin, a (90)Y radiolabeled murine antibody. Both target CD20 receptors present on the surface of lymphocytes. We present our clinical experience with Bexxar and Zevalin in the management of low-grade refractory or relapsed NHL. METHODS: This is a retrospective study (Jan 2000-Jul 2006) of 67 patients with NHL, who were treated with Bexxar (31 patients, group A) or Zevalin (36 patients, group B) for refractory/relapsed disease. Group A included 16 men and 15 women, 35-81 years old (average, 59.3 +/- 13.4). Group B included 27 men and nine women, 36-85 years old (average, 55.4 +/- 13.8). Therapeutic doses ranged 40-138 mCi (average, 78.1 +/- 28.2) for Bexxar and 17-34 mCi (average, 28.8 +/- 4.37) for Zevalin. RESULTS: Objective responses were induced in 22 of the 31 patients (70.9%) in group A and 28 of the 36 patients (77.8%) in group B. Complete response was noted in 11 patients (35.5%), partial response in seven patients (22.6%), and mixed response in four patients (12.9%) in group A. There were five patients (16.1%) with stable disease and four patients (12.9%) with disease progression in the same group. Complete response was noted in 15 patients (41.7%), partial response in nine patients (25%), and mixed response in four patients (11.1%) in group B. There were four patients (11.1%) with stable disease and another four patients (11.1%) with disease progression in the same group. The average decreases at posttherapy nadir were 36.9% +/- 0.33 (group A) and 52.6% +/ 0.32 (group B) for platelets, 27.8% +/- 0.27 (group A) and 34.2% +/- 0.38 (group B) for leukocytes, and 4.9% +/- 0.15 (group A) and 7.6% +/- 0.11 (group B) for hemoglobin. Grades 3 and 4 hematological toxicity occurred in 14 patients (45.2%) treated with Bexxar and 22 patients (61.1%) treated with Zevalin, but was reversible. CONCLUSION: Our study suggests that clinical practice of Bexxar and Zevalin radioimmunotherapy is an effective and safe adjunctive treatment for patients with NHL refractory/relapsed to conventional treatment. However, due to the small number of subjects, it was not possible to determine whether differences in the outcomes or toxicities from the two agents were statistically significant. PMID- 19543948 TI - Rediscovering the individual in mediation analyses. AB - Using the article by von Eye and colleagues as a springboard, this commentary considers a variety of challenges to valid exploration of mediation processes. Along the way, a several alternatives for reinserting the individual back into mediation processes are entertained. The configural frequency approach to mediation analyses proposed by von Eye et al. (Integrative Psychological & Behavioral Science 43(3), 2009) represents a major step toward this objective. Idiographic approaches are proposed as the next step in the progression toward individual-centered mediation analyses. PMID- 19543947 TI - Incremental value of integrated FDG-PET/CT in evaluating indeterminate solitary pulmonary nodule for malignancy. AB - OBJECTIVES: The objective of this study was to evaluate the increased diagnostic benefit of integrated positron emission tomography/computed tomography (PET/CT) interpretation in evaluating solitary pulmonary nodules for malignancy. METHODS: One hundred seventeen patients (67 men and 50 women; mean age +/- SD, 61.7 +/- 13.6 years, range, 31-86 years) with indeterminate solitary pulmonary nodules and no previous history of malignancy were analyzed. PET/CT was performed with an integrated PET/CT scanner (Siemens Biograph BGO duo) 1 h after an intravenous injection of 370 MBq (10 mCi) (18)F-fluorodeoxyglucose. Patients fasted for 6 h before imaging. PET was interpreted alone or combined with CT and was graded according to a five-point scale. A malignant diagnosis was based on histological findings or a clinical and radiological follow-up after at least 24 months. The diagnostic performances of PET alone and integrated PET/CT interpretation were evaluated using discriminant analysis. RESULTS: PET alone correctly classified 85% of nodules and integrated PET/CT interpretation increased the correct classification to 89%, with similar sensitivity and specificity of 88% and 89%, respectively. False-positive PET results mainly resulted from granulomatous disorders. Four (50%) of the eight cases deemed indeterminate on PET alone were resolved with combined PET/CT interpretation. CONCLUSIONS: Although the benefit attributable to the CT component was limited when integrated PET/CT was used, PET and CT acted synergistically to significantly increase the diagnostic veracity for PET-indeterminate nodules. PMID- 19543949 TI - An unusual cyanobacterium from saline thermal waters with relatives from unexpected habitats. AB - Cyanobacteria that grow above seawater salinity at temperatures above 45 degrees C have rarely been studied. Cyanobacteria of this type of thermo-halophilic extremophile were isolated from siliceous crusts at 40-45 degrees C in a geothermal seawater lagoon in southwest Iceland. Iceland Clone 2e, a Leptolyngbya morphotype, was selected for further study. This culture grew only at 45-50 degrees C, in medium ranging from 28 to 94 g L(-1) TDS, It showed 3 doublings 24 h(-1) under continuous illumination. This rate at 54 degrees C was somewhat reduced, and death occurred at 58 degrees C. A comparison of the 16S rDNA sequence with all others in the NCBI database revealed 2 related Leptolyngbya isolates from a Greenland hot spring (13-16 g L(-1) TDS). Three other similar sequences were from Leptolyngbya isolates from dry, endolithic habitats in Yellowstone National Park. All 6 formed a phylogenetic clade, suggesting common ancestry. These strains shared many similarities to Iceland Clone 2e with respect to temperature and salinity ranges and optima. Two endolithic Leptolyngbya isolates, grown previously at 23 degrees C in freshwater medium, grew well at 50 degrees C but only in saline medium. This study shows that limited genotypic similarity may reveal some salient phenotypic similarities, even when the related cyanobacteria are from vastly different and remote habitats. PMID- 19543950 TI - The evolution of self-compatible and self-incompatible populations in a hermaphroditic perennial, Trillium camschatcense (Melanthiaceae). AB - The evolution of selfing from outcrossing ancestors is known to have occurred repeatedly in angiosperms. Theoretical studies have argued that the transition from outcrossing to selfing is accomplished more easily than the reverse case, and phylogenetic analyses involving self-compatible (SC) and self-incompatible (SI) species has basically supported this assumption. The evolutionary direction of self-compatibility and self-incompatibility was examined in Trillium camschatcense, which contains geographically widespread SC populations, and restricted SI populations. Ecological surveys have revealed that the SC populations were suitable for outcrossing, and selfing in these populations did not confer any fitness advantage. Since reproductive fitness indicates the possibility of an evolutionary shift from self-compatibility to self incompatibility, the phylogenetic relationships of SI and SC populations of T. camschatcense were investigated based on cpDNA variations and nuclear DNA microsatellite polymorphisms. Although phylogenetic analyses did not provide credible evidence to determine evolutionary direction, the SI populations turned out to be monophyletic with extremely low genetic differentiation. Based on these results, we proposed two possible scenarios for the evolutionary backgrounds of SI and SC populations in T. camschatcense. The plausibility of each scenario was evaluated based on the reproductive and geographical features of the mating systems. PMID- 19543951 TI - Rituximab for the treatment of corticosteroid-refractory chronic graft-versus host disease. AB - We prospectively evaluated the safety and efficacy of the anti-CD20 chimeric monoclonal antibody rituximab for the treatment of corticosteroid-refractory chronic graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation. Seven patients were treated with 375 mg/m(2) rituximab weekly for 4 consecutive weeks. Rituximab was well tolerated with no severe toxicity observed during treatment. At 1 year, 3 patients showed a partial response to rituximab therapy, 3 had stable disease, and 1 had progressive disease. Rituximab allowed a reduction in the dose of steroids in 4 patients. Responsive manifestations included mild to moderate skin and oral lesions, and immune hemolytic anemia, and thrombocytopenia. Severe manifestations involving the skin, fascia, and eye did not respond to treatment. These observations suggest that rituximab therapy may be effective for select patients with corticosteroid refractory chronic GVHD that is not advanced. PMID- 19543952 TI - A phase II, open-label, sequential-cohort, dose-escalation study of romiplostim in Japanese patients with chronic immune thrombocytopenic purpura. AB - This phase II, multicenter, open-label, sequential-cohort, dose-escalation study was designed to evaluate the safety and efficacy of romiplostim, a novel peptibody that increases platelet production, in Japanese patients with chronic immune thrombocytopenic purpura (ITP). Sequential cohorts of four patients each received romiplostim (1, 3, or 6 microg/kg) subcutaneously on days 1 and 8 of the dose-escalation phase. Patients who achieved platelet responses (doubling of baseline platelet counts to > or =50 x 10(9)/L) continued romiplostim weekly during the treatment-continuation phase. Romiplostim produced dose-dependent increases in mean and peak platelet counts. Five patients received romiplostim during the treatment-continuation phase, with platelet counts > or =50 x 10(9)/L maintained in approximately half of the weekly assessments. Romiplostim was well tolerated. No severe, serious, or life-threatening adverse events were reported. No binding antibodies to romiplostim or thrombopoietin were detected. Romiplostim is safe and well tolerated in Japanese patients with chronic ITP and is effective in producing platelet count increases, consistent with the results from studies in non-Japanese patients. On the basis of these findings, a starting dose of 3 microg/kg was recommended for phase III evaluation of romiplostim in Japanese patients with chronic ITP. PMID- 19543953 TI - Nail dystrophy due to dose-intensive chemotherapy. PMID- 19543954 TI - Abnormal IgD and IgA1 O-glycosylation in hyperimmunoglobulinaemia D and periodic fever syndrome. AB - In order to determine the glycosylation pattern for IgD, and to examine whether there are changes in the pattern of IgD and IgA1 O-glycosylation in patients with hyperimmunoglobulinaemia D and periodic fever syndrome (HIDS) during acute febrile attacks and during periods of quiescence, serum was obtained from 20 patients with HIDS and 20 control subjects. In the HIDS group, serum was obtained either during an acute febrile episode (n = 9) or during a period of quiescence (n = 11). The O-glycosylation profiles of native and desialylated IgA1 and IgD were measured in an ELISA-type system using the lectins Helix aspersa and peanut agglutinin, which bind to alternative forms of O-glycan moieties. IgD is more heavily O-galactosylated and less O-sialylated than IgA1 in healthy subjects. HIDS is associated with more extensive O-galactosylation of IgD and a reduction in O-sialylation of both IgD and IgA1. These changes are present both during acute febrile attacks and periods of quiescence. The T cell IgD receptor is a lectin with binding affinity for the O-glycans of both IgD and IgA1. The observed changes in IgD and IgA1 O-glycosylation are likely to have a significant effect on IgD/IgA1-T cell IgD receptor interactions including basal immunoglobulin synthesis, and possibly myeloid IgD receptor-mediated cytokine release. PMID- 19543957 TI - Charge transfer photophysics of tetra(alpha-amino) zinc phthalocyanine. AB - The absorption, fluorescence, and transient absorption spectra of Tetra(alpha amino) zinc phthalocyanine, ZnPc(alpha-NH2)4, have been measured in polar solvents and compared with that of ZnPc(alpha-R)4 (R = H, NO2, OCH(CH3)2). While the latter three showed the typical photophysics of phthalocyanines, ZnPc(alpha NH2)4 exhibits distinct spectral properties, a very low fluorescence quantum yield and a relatively long fluorescence lifetime. These observations are explained by the substantial charge transfer characters in the absorption and fluorescence spectra of ZnPc(alpha-NH2)4. NMR indicates that intramolecular H bonding makes atoms in NH2 actually coplanar with other elements of ZnPc(alpha NH2)4. The local excited state is non emissive and the weak emission is assigned to its charge transfer state. The transient absorption bands from laser flash photolysis located at 630 nm, 645 nm is assigned to the mono-charge transfer state, while that at 545 nm is assigned to the di-charge transfer state. PMID- 19543956 TI - Hydrogel matrix entrapping PLGA-paclitaxel microspheres: drug delivery with near zero-order release and implantability advantages for malignant brain tumour chemotherapy. AB - PURPOSE: To develop paclitaxel-delivering PLGA microspheres entrapped in a gel matrix with sustained drug release properties and implantability advantages for local glioma chemotherapy. METHODS: Paclitaxel-loaded PLGA microspheres were fabricated using electrohydrodynamic atomization and entrapped by electrospray and gelation. The physicochemical characterizations were performed using scanning electron microscopy and differential scanning calorimetry. The influence of various parameters on the disintegration time was investigated. In vitro release of paclitaxel was quantified using high performance liquid chromatography. Cytotoxicity of the formulations was assessed by the quantification of IC(50) and caspase-3 activity against C6 glioma cells in vitro. The formulations were tested against a subcutaneous C6 glioma tumour in mice. RESULTS: Highly monodisperse gel beads containing a uniform microsphere distribution were obtained. Gelation using Ca(2+) ions ensured entrapment of microspheres with high loading efficiency. With an increase in the gelation time, gelling bath concentration and decrease in microsphere loading, it was more difficult to disintegrate the beads and release the microspheres. The formulations demonstrated sustained drug release for more than 60 days at a near-constant rate and a low initial burst. Cell culture studies proved the cytotoxicity against C6 glioma and improved performance in comparison to Taxol. The formulations could reduce subcutaneous tumour volume to a greater extent compared to Taxol and the control. CONCLUSIONS: Paclitaxel loaded PLGA microspheres entrapped in an alginate gel matrix could be potential local chemotherapy implants to treat malignant glioma with critical advantages of implantability and sustained drug release with low initial burst. PMID- 19543955 TI - Gallic acid, an active constituent of grape seed extract, exhibits anti proliferative, pro-apoptotic and anti-tumorigenic effects against prostate carcinoma xenograft growth in nude mice. AB - PURPOSE: Gallic acid, a natural agent present in a wide-range of fruits and vegetables, has been of potential interest as an anti-cancer agent; herein, we evaluated its efficacy in androgen-independent DU145 and androgen-dependent-22Rv1 human prostate cancer (PCa) cells. MATERIALS AND METHODS: Cell viability was determined by MTT and apoptosis by Annexin V-PI assays. In vivo anti-cancer efficacy was assessed by DU145 and 22Rv1 xenograft growth in nude mice given normal drinking water or one supplemented with 0.3% or 1% (w/v) gallic acid. PCNA, TUNEL and CD31 immunostaining was performed in tumor tissues for in vivo anti-proliferative, apoptotic and anti-angiogenic effects of gallic acid. RESULTS: Gallic acid decreased cell viability in a dose-dependent manner in both DU145 and 22Rv1 cells largely via apoptosis induction. In tumor studies, gallic acid feeding inhibited the growth of DU145 and 22Rv1 PCa xenografts in nude mice. Immunohistochemical analysis revealed significant inhibition of tumor cell proliferation, induction of apoptosis, and reduction of microvessel density in tumor xenografts from gallic acid-fed mice as compared to controls in both DU145 and 22Rv1 models. CONCLUSION: Taken together, our findings show the anti-PCa efficacy of gallic acid and provide a rationale for additional studies with this naturally-occurring agent for its efficacy against PCa. PMID- 19543958 TI - Specific immunotherapy to birch allergen does not enhance suppression of Th2 cells by CD4(+)CD25(+) regulatory T cells during pollen season. AB - INTRODUCTION: The aim of this study was to investigate the suppressive capacity of CD25(+) regulatory T cells on birch allergen-induced T-cell responses during the first birch pollen season after initiation of specific immunotherapy (SIT). METHODS: CD25(pos) and CD25(neg) T cells were purified from blood of birch allergic SIT patients and birch-allergic controls, stimulated with birch pollen extract, and analyzed for T-cell proliferation and production of interferon gamma (IFN-gamma), interleukin (IL)-5 and IL-10. RESULTS: We show that allergen-induced proliferation and IFN-gamma production were suppressed equally well by CD25(pos) T cells from SIT patients and controls, while the IL-5 production was not suppressed by either of the groups. IL-10 levels were higher in SIT patients relative to controls only when CD25(neg) and CD25(pos) were cultured together. Furthermore, neither FOXP3 levels nor proportions of CD25(high) T cells were enhanced in SIT patients compared to allergic controls. DISCUSSION: These results suggest that the Th2-suppressive capacity of allergen-stimulated CD25(pos) Treg in vitro is not improved by SIT in spite of increased IL-10 production from T cells. PMID- 19543959 TI - IL-2, IFN-gamma, and IL-12 gene polymorphisms and susceptibility to multiple sclerosis. AB - BACKGROUND: Multiple sclerosis (MS) is a multifactorial disease. Positive genetic background could predispose individuals to this chronic disabling disease. In order to investigate the role of some proinflammatory cytokines (interleukin (IL) 2, IL-12, and interferon-gamma (IFN-gamma)) as a risk factor for MS, this study was performed. METHODS: Two hundred and eleven patients with relapsing-remitting form of MS were enrolled in this study and compared with 359 healthy individuals. Using polymerase chain reaction based on sequence-specific primer method, the cytokine genes were amplified, and alleles and genotypes were detected on gel electrophoresis. RESULTS: Significant increases for IFN-gamma AT (+874) genotype (54.5% vs. 37.8%, p = 0.0002) and IL-12 AA (-1188) genotype (60.8% vs. 49.7%, p = 0.014) were found in MS patients in comparison with healthy controls. A significant decrease in IFN-gamma TT (+874) genotype (17.7% vs. 27.5%, p = 0.01) and IL-12 CA (-1188) genotype (30.9% vs. 45%, p = 0.001) in MS patients was also detected. No significant differences of IL-2 G/T (-330) and IL-2 G/T (+166) in alleles and genotypes were observed between MS patients and normal subjects. CONCLUSIONS: It could be suggested that the genetic variation in IL-12 A/C ( 1188) and IFN-gamma A/T (+874) cytokine genes could be risk factors for MS patients. PMID- 19543961 TI - Endovascular bi-ventricular pacing-defibrillator placement using a trans-atrial approach. AB - Venous access for pectoral pacemaker and defibrillator lead placement can be compromised by venous occlusion due to previous pacing leads, access ports for medications such as chemotherapy, dialysis access, and other causes. On rare occasion, a femoral access is utilized for device placement. We report here a patient without venous access to the heart from either above or below due to retroperitoneal fibrosis. A bi-ventricular pacing-defibrillator was placed using a direct trans-atrial approach with good results. This minimally invasive approach to device placement may be useful in patients with poor venous access and avoids the placement of epicardial hardware. PMID- 19543960 TI - Modulation of the proteome of peripheral blood mononuclear cells from HIV-1 infected patients by drugs of abuse. AB - INTRODUCTION: We used proteomic analyses to assess how drug abuse modulates immunologic responses to infections with the human immunodeficiency virus type 1 (HIV-1). METHODS: Two-dimensional difference gel electrophoresis was utilized to determine changes in the proteome of peripheral blood mononuclear cells (PBMC) isolated from HIV-1-positive donors that occurred after treatment with cocaine or methamphetamine. Both drugs differentially regulated the expression of several functional classes of proteins. We further isolated specific subpopulations of PBMC to determine which subpopulations were selectively affected by treatment with drugs of abuse. Monocytes, B cells, and T cells were positively or negatively selected from PBMC isolated from HIV-1-positive donors. RESULTS: Our results demonstrate that cocaine and methamphetamine modulate gene expression primarily in monocytes and T cells, the primary targets of HIV-1 infection. Proteomic data were validated with quantitative, real-time polymerase chain reaction. These studies elucidate the molecular mechanisms underlying the effects of drugs of abuse on HIV-1 infections. Several functionally relevant classes of proteins were identified as potential mediators of HIV-1 pathogenesis and disease progression associated with drug abuse. PMID- 19543962 TI - Increased levels of 25 hydroxyvitamin D and 1,25-dihydroxyvitamin D after rosuvastatin treatment: a novel pleiotropic effect of statins? AB - OBJECTIVES: Low levels of 25-hydroxyvitamin D are associated with higher risk of cardiovascular morbidity and mortality. Large trials demonstrated that statins significantly decrease cardiovascular morbidity and mortality. 7 dehydrocholesterol is the precursor of both cholesterol and vitamin D. The aim of this study was to investigate the possible effect of rosuvastatin on vitamin D metabolism. METHODS: The study was performed in a prospective cohort design. The study group consisted of 91 hyperlipidemic patients who had not been treated with lipid lowering medications. Lipid parameters, 25 hydroxyvitamin-D, 1,25 dihydroxyvitamin D, and bone alkaline phosphatase were obtained at baseline and after 8 weeks of rosuvastatin treatment. RESULTS: None of the subjects withdrew from the study because of the adverse effects. The mean age was 59.9 +/- 12.5 years. The majority of the patients were male (55, 60%). Seventeen patients were diabetic, and 43 patients had systemic hypertension. There was a significant increase in 25-hydroxyvitamin D, from mean 14.0 (range 3.7- 67) to mean 36.3 (range 3.8 -117) ng/ml (p < 0.001), and also an increase of 1,25-dihydroxyvitamin D from mean 22.9 +/- 11.2 to 26.6 +/- 9.3 pg/dl (p = 0.023). Bone alkaline phosphatase decreased after 8 weeks of rosuvastatin treatment, mean 17.7 (range 2.6-214) to mean 9.5 (range 2.3-19.1) u/l (p < 0.001) rosuvastatin treatment. CONCLUSION: This study has shown an effect of rosuvastatin on vitamin D metabolism, with an increase in both 25-hydroxyvitamin D and 1,25 dihydroxyvitamin D. This may be an important pleiotropic effect whereby rosuvastatin reduces mortality in patients with coronary artery disease. Further studies are needed to clarify the relationship between statins and vitamin D metabolism. PMID- 19543963 TI - Variability of bursting patterns in a neuron model in the presence of noise. AB - Spiking and bursting patterns of neurons are characterized by a high degree of variability. A single neuron can demonstrate endogenously various bursting patterns, changing in response to external disturbances due to synapses, or to intrinsic factors such as channel noise. We argue that in a model of the leech heart interneuron existing variations of bursting patterns are significantly enhanced by a small noise. In the absence of noise this model shows periodic bursting with fixed numbers of interspikes for most parameter values. As the parameter of activation kinetics of a slow potassium current is shifted to more hyperpolarized values of the membrane potential, the model undergoes a sequence of incremental spike adding transitions accumulating towards a periodic tonic spiking activity. Within a narrow parameter window around every spike adding transition, spike alteration of bursting is deterministically chaotic due to homoclinic bifurcations of a saddle periodic orbit. We have found that near these transitions the interneuron model becomes extremely sensitive to small random perturbations that cause a wide expansion and overlapping of the chaotic windows. The chaotic behavior is characterized by positive values of the largest Lyapunov exponent, and of the Shannon entropy of probability distribution of spike numbers per burst. The windows of chaotic dynamics resemble the Arnold tongues being plotted in the parameter plane, where the noise intensity serves as a second control parameter. We determine the critical noise intensities above which the interneuron model generates only irregular bursting within the overlapped windows. PMID- 19543964 TI - Expression of porcine parvovirus VP2 gene requires codon optimized E. coli cells. AB - Porcine parvovirus (PPV) is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. The gene coding for the capsid protein VP2 of PPV was amplified and inserted into the plasmid pET-32a (+), which was then used to transform Escherichia coli Rosetta, the capsid protein of PPV was fused to a polyhistidine tag, and the position of the affinity tag is in N-terminus. VP2 was expressed using different expression host bacteria, including E. coli BL21, and Rosetta, and different plasmid vectors, including pET 30a (+), pET-32a (+), and pGEX-6p-1. After selection, only the fusion protein inserted into pET-32a (+) was expressed well in E. coli Rosetta. The recombinant bacterium produced high quantities of the fusion protein VP2, about 8% in total. The expressed VP2 was antigenically similar to the native capsid protein according to a Western blot assay performed with polyclonal antibodies obtained from pigs vaccinated with PPV. A simple, easily commercialized procedure was used to purify this protein. This study provides a foundation for the application of VP2 protein in the clinical diagnosis of PPV and in the vaccination against PPV. PMID- 19543965 TI - Comment on Winke T et al. J Assist Reprod Genet DOI 10.1007/S10815-008-9277-3. PMID- 19543966 TI - Prediction of high ovarian response to controlled ovarian hyperstimulation: anti Mullerian hormone versus small antral follicle count (2-6 mm). AB - PURPOSE: To evaluate the predictive value of basal serum anti-mullerian hormone level and small antral follicle count for high ovarian response to controlled ovarian hyperstimulation. METHODS: A total of 159 patients were prospectively included. Basal serum anti-mullerian hormone and small antral follicle count (2-6 mm) were measured. RESULTS: Small antral follicle count and anti-mullerian hormone have similar predictive accuracy for high ovarian response with area under curve of 0.961 and 0.922, respectively. The sensitivity and specificity for prediction of high ovarian response were 89% and 92% for small antral follicle count and 93% and 78% for anti-mullerian hormone at the cutoff values of > or = 16 and > or = 34.5 pmol/l, respectively. CONCLUSIONS: Small antral follicle count and anti-mullerian hormone are equally accurate predictors of high ovarian response and facilitate determination of the optimal strategy for controlled ovarian hyperstimulation. PMID- 19543969 TI - Genetic diversity and genetic structure analysis of the natural populations of Lilium brownii from Guangdong, China. PMID- 19543968 TI - Re-examining the epidemiology of sensory regulation dysfunction and comorbid psychopathology. AB - The aims of this study were threefold: (1) to compare prevalence of sensory regulation dysfunction based on previously established criteria to rates established with a more representative community sample of 796 4-year-olds; (2) to examine ethnic/racial and gender differences in prevalence according to the different criteria; and (3) to examine the co-occurrence of sensory regulation dysfunction and preschool psychiatric disorders. Prevalence rates ranged from 3.4% (current criteria) to 15.6% (previous criteria). In contrast to previous studies with less representative samples, there were no significant ethnic or racial differences using the current criteria. Boys were more likely to have sensory regulation dysfunction than girls according to all criteria. Depending upon impairment criteria used, 33-63% of children meeting criteria for sensory dysregulation also had a psychiatric disorder; 37-67% had only a sensory dysregulation disorder, indicating that sensory regulation dysfunction exists independent of psychiatric disorder, and is also a significant risk factor for disorder. PMID- 19543967 TI - Inhibition, reinforcement sensitivity and temporal information processing in ADHD and ADHD+ODD: evidence of a separate entity? AB - This study compared children with ADHD-only, ADHD+ODD and normal controls (age 8 12) on three key neurocognitive functions: response inhibition, reinforcement sensitivity, and temporal information processing. The goal was twofold: (a) to investigate neurocognitive impairments in children with ADHD-only and children with ADHD+ODD, and (b) to test whether ADHD+ODD is a more severe from of ADHD in terms of neurocognitive performance. In Experiment 1, inhibition abilities were measured using the Stop Task. In Experiment 2, reinforcement sensitivity and temporal information processing abilities were measured using a Timing Task with both a reward and penalty condition. Compared to controls, children with ADHD only demonstrated impaired inhibitory control, showed more time underestimations, and showed performance deterioration in the face of reward and penalty. Children with ADHD+ODD performed in-between children with ADHD-only and controls in terms of inhibitory controls and the tendency to underestimate time, but were more impaired than controls and children with ADHD-only in terms of timing variability. In the face of reward and penalty children with ADHD+ODD improved their performance compared to a neutral condition, in contrast to children with ADHD-only. In the face of reward, the performance improvement in the ADHD+ODD group was disproportionally larger than that of controls. Taken together the findings suggest that, in terms of neurocognitive functioning, comorbid ADHD+ODD is a substantial different entity than ADHD-only. PMID- 19543970 TI - Assigned versus random, countermeasure-like responses in the p300 based complex trial protocol for detection of deception: task demand effects. AB - We recently introduced an accurate and countermeasure resistant P300-based deception detection test called the complex trial protocol (Rosenfeld et al. in Psychophysiology 45(6):906-919, 2008). When subjects use countermeasures to all irrelevant items in the test, the probe P300 is increased rather than reduced (as it was in previous P300-based deception protocols), allowing detection of countermeasure users. The current experiment examines the role of task demand on the complex trial protocol by forcing the subject to make countermeasure-like response to stimuli. Subjects made either a simple random button response to both probe and irrelevant stimuli (experiment 1) or a more complex, assigned, button response to probe and irrelevant stimuli (experiment 2). We found that an increase in task demand reduced the effectiveness of the test. Using random responses we found a simple guilty hit rate of 11/12 with no false positives, but only a 4/11 hit rate for countermeasure-users. Using assigned responses we found a simple guilty hit rate of 8/15 with no false positives, and a 7/16 hit rate for countermeasure-users. We herein suggest that the high level of task demand associated with these countermeasure-like responses causes reduced hit rates. PMID- 19543971 TI - Is red meat intake a risk factor for breast cancer among premenopausal women? AB - Breast cancer is the second leading cause of cancer deaths in women today and is the most common cancer among women. Although a number of risk factors such as genetics, family history, parity, age at first birth, and age at menarche and menopause have been established, most are difficult to modify. Diet, however, is a potentially modifiable approach for prevention and a variety of dietary patterns have been examined with respect to their role in breast cancer. One such dietary factor is red meat consumption. Red meat intake has been hypothesized to increase breast cancer risk but while both case-control and ecologic studies have supported a positive association, prospective cohort studies have been inconsistent. One explanation for this inconsistency may be related to menopausal status. We performed a meta-analysis on the association between breast cancer risk and red meat consumption in premenopausal women. A total of ten studies were identified. The summary relative risk was 1.24 (95% CI 1.08-1.42). Case-control studies (N = 7) had a risk of 1.57 (95% CI 1.23-1.99), while cohort studies (N = 3) had a summary relative risk of 1.11 (95% CI 0.94-1.31). PMID- 19543972 TI - Characterization of BRCA1 ring finger variants of uncertain significance. AB - The majority of pathogenic mutations in BRCA1 result in a truncated protein. Although most missense changes in BRCA1 are of unknown functional significance, a handful of deleterious missense mutations have been identified. The majority of these occur in splice sites or highly conserved protein domains. Previously, we developed a predictive model, VUS Predict, to classify BRCA variants of uncertain significance as neutral or deleterious. It uses evolutionary prediction algorithms together with clinical information from cancer pathology reports and BRCA genetic testing results. Because of the higher probability that missense changes occurring in conserved BRCA1 domains are of pathogenic significance, we identified all individuals in our cohort who had been tested for BRCA1 and BRCA2 mutations who had missense changes in the BRCA1 ring finger domain and sought to classify those changes. We applied VUS Predict to three previously uncharacterized variants and four missense changes known to be deleterious. Two variants, L22S and T37K, were predicted to be deleterious and one variant, K45Q, was predicted to be neutral by VUS Predict. The mutations C39R, C44Y, C44S, and C61G were confirmed as deleterious. PMID- 19543973 TI - Impact of menopausal symptoms on quality of life 6 months after systemic breast cancer treatment: results from the Shanghai Breast Cancer Survival Study. AB - Menopausal symptoms (MPS) after breast cancer treatment are associated with reduced health-related quality of life (QOL) among Caucasian women. Little is known about whether MPS similarly impact QOL in Asian women with breast cancer. QOL was assessed by using the generic quality of life instruments, Medical Outcome Study of Quality of Life Inventory (MOSQOL-74) or Short Form-36 Health Survey (SF-36) in 4,976 Chinese participants of the Shanghai Breast Cancer Survival Study who were treated for incident, non-metastatic breast cancer within the 6 months before the study interview. Relationships between MPS and QOL were assessed by multiple linear regression, controlling for potential confounders. About 71.4% of study participants experienced MPS, including hot flashes, night sweats, vaginal dryness, depressed mood, and/or dry skin. Women with MPS reported lower overall QOL than women without MPS [mean scores 61.0 vs. 64.0, respectively (MOSQOL-74) and 54.9 vs. 66.9, respectively (SF-36); P < 0.01]. Adjusted mean differences (beta) in overall QOL in the presence and absence of MPS were -3.1 (95% CI -3.8, -2.4) with the MOSQOL-74 and -12.3 (95% CI -13.8, -10.9) with the SF-36. Women with any MPS had lower scores for the MOSQOL-74 physical and psychological domains and for the SF-36 social and emotional subscales than those without MPS (P < 0.05 for all). Having several MPS predicted poorer QOL in all domains measured regardless of the instrument used (P (trend) < 0.01 for all). Our study indicates that in Chinese women recently treated for breast cancer, MPS adversely impacts QOL. Actively soliciting and treating MPS in these women should significantly improve their QOL. PMID- 19543975 TI - Quantitative prediction of body surface potentials from myocardial action potentials using a summed dipole model. AB - This paper demonstrates quantitatively, using streamlined mathematics, how the transmembrane ionic currents in individual cardiac muscle cells act to produce the body surface potentials of the electrocardiogram (ECG). From fundamental principles of electrostatics, anatomy, and physiology, one can characterize the strength of apparent dipoles along a wavefront of depolarization in a local volume of myocardium. Net transmembrane flow of ionic current in actively depolarizing or repolarizing tissue induces extracellular current flow, which sets up a field of electrical potential that resembles that of a dipole. The local dipole strength depends upon the tissue cross section, the tissue resistivity, the resting membrane potential, the membrane capacitance, the volume fraction of intracellular fluid, the time rate of change of the action potential, and the cell radius. There are no unknown, "free" parameters. There are no arbitrary scale factors. Body surface potentials are a function of the summed local dipole strengths, directions, and distances from the measuring points. Calculations of body surface potentials can be made for the scenarios of depolarization (QRS complex), repolarization (T wave) and localized acute injury (ST segment shifts) and agree well with experimentally measured potentials. This simplified predictive dipole theory provides a solution to the forward problem of electrocardiography that explains from a physiological perspective how the collective depolarization and repolarization of individual cardiac muscle cells create body surface potentials in health and disease. PMID- 19543976 TI - BCL2DB: moving 'helix-bundled' BCL-2 family members to their database. PMID- 19543974 TI - Perceived mental illness stigma and HIV risk behaviors among adult psychiatric outpatients in Rio de Janeiro, Brazil. AB - We examined the associations between perceived mental illness stigma and HIV risk and protective behaviors among adults with severe mental illness (SMI) in Rio de Janeiro, Brazil. We measured mental illness stigma across three domains ("Personal Experiences," "Perceived Attractiveness," and "Relationship Discrimination"), and examined the relationship between experiences of stigma in each domain and HIV risk and protective behaviors over the past 3 months in 98 outpatients with SMI. Those who reported greater "Relationship Discrimination" stigma were significantly more likely to be sexually active and to have unprotected sex; they were significantly less likely to report deliberately having fewer partners as a way to protect themselves from HIV. The role of stigma in unprotected sexual behavior should be examined further and considered in any HIV prevention intervention for people with SMI. PMID- 19543977 TI - Sexual Acquisition and Transmission of HIV Cooperative Agreement Program (SATHCAP), July 2009: introduction. PMID- 19543978 TI - Refined homology model of monoacylglycerol lipase: toward a selective inhibitor. AB - Monoacylglycerol lipase (MGL) is primarily responsible for the hydrolysis of 2 arachidonoylglycerol (2-AG), an endocannabinoid with full agonist activity at both cannabinoid receptors. Increased tissue 2-AG levels consequent to MGL inhibition are considered therapeutic against pain, inflammation, and neurodegenerative disorders. However, the lack of MGL structural information has hindered the development of MGL-selective inhibitors. Here, we detail a fully refined homology model of MGL which preferentially identifies MGL inhibitors over druglike noninhibitors. We include for the first time insight into the active site geometry and potential hydrogen-bonding interactions along with molecular dynamics simulations describing the opening and closing of the MGL helical-domain lid. Docked poses of both the natural substrate and known inhibitors are detailed. A comparison of the MGL active-site to that of the other principal endocannabinoid metabolizing enzyme, fatty acid amide hydrolase, demonstrates key differences which provide crucial insight toward the design of selective MGL inhibitors as potential drugs. PMID- 19543979 TI - Identification of family-specific residue packing motifs and their use for structure-based protein function prediction: I. Method development. AB - Protein function prediction is one of the central problems in computational biology. We present a novel automated protein structure-based function prediction method using libraries of local residue packing patterns that are common to most proteins in a known functional family. Critical to this approach is the representation of a protein structure as a graph where residue vertices (residue name used as a vertex label) are connected by geometrical proximity edges. The approach employs two steps. First, it uses a fast subgraph mining algorithm to find all occurrences of family-specific labeled subgraphs for all well characterized protein structural and functional families. Second, it queries a new structure for occurrences of a set of motifs characteristic of a known family, using a graph index to speed up Ullman's subgraph isomorphism algorithm. The confidence of function inference from structure depends on the number of family-specific motifs found in the query structure compared with their distribution in a large non-redundant database of proteins. This method can assign a new structure to a specific functional family in cases where sequence alignments, sequence patterns, structural superposition and active site templates fail to provide accurate annotation. PMID- 19543982 TI - Effect of different operational conditions on the decolorization of molasses spent wash using once developed soil inoculum. AB - A stirred vessel coupled with membrane unit containing cellulose acetate (0.45 mum) membrane was used to study the decolorization of anaerobically digested molasses spent wash (MSW). The soil collected from the MSW disposal site was used as inoculum to study the decolorization without addition of any additives. The same inoculum was used over a period of 163 days at room temperature to study the decolorization of 12.5-50% (v/v) MSW using different operational conditions. The reactor was entered in to the inhibition mode after the feeding of 50% MSW, which was restored 100% without changing any operational condition. The maximum decolorization obtained for 12.5% (v/v) MSW was 77.22 +/- 0.13%. The decolorization achieved for 25, 37.5, and 50% (v/v) MSW was 70.41 +/- 0.12, 56.47 +/- 0.17, and 48.78 +/- 0.09%, respectively. Increase in the utilization of protein and reducing sugar was observed up to 25% MSW whereas, higher concentration showed decrease in the utilization. Results indicate 63% removal of chemical oxygen demand for 12.5% (v/v) MSW. Membrane flux which was significantly reduced after the feeding of 50% MSW was regenerated without changing the washing procedure, however, 35% decrease in sample flux was observed over the continuous use of membrane for the period of 198 days. PMID- 19543981 TI - Karyotypic characterization of Trachemys dorbigni (Testudines: Emydidae) and Chelonoidis (Geochelone) donosobarrosi (Testudines: Testudinidae), two species of Cryptodiran turtles from Argentina. AB - We describe for the first time the karyotypes of two species of Cryptodiran turtles from Argentina, namely, Trachemys dorbigni (Emydidae) and Chelonoidis (Geochelone) donosobarrosi (Testudinidae). The karyotype of T. dorbigni (2n = 50) consists of 13 pairs of macrochromosomes and 12 pairs of microchromosomes, whereas the karyotype of C. donosobarrosi (2n = 52) consists of 11 pairs of macrochromosomes and 15 pairs of microchromosomes. Fluorescence in situ hybridization (FISH) with a (TTAGGG)n telomeric probe showed that the chromosomes of these species have four telomeric signals, two at each end, indicating that none of the chromosomes of T. dorbigni and C. donosobarrosi are telocentric. The fact that no interstitial telomeric signals were observed after FISH, suggests that interstitial telomeric sequences did not have a major role in the chromosomal evolution of these species. Additional data will be needed to elucidate if interstitial telomeric sequences have a major role in the karyotypic evolution of Testudines. PMID- 19543980 TI - Homospermidine in transgenic tobacco results in considerably reduced spermidine levels but is not converted to pyrrolizidine alkaloid precursors. AB - Homospermidine synthase is the first specific enzyme in the biosynthesis of pyrrolizidine alkaloids. Whereas the substrates putrescine and spermidine are part of the highly dynamic polyamine pool of plants, the product homospermidine is incorporated exclusively into the necine base moiety of pyrrolizidine alkaloids. Recently, the gene encoding homospermidine synthase has been shown to have been recruited several times independently during angiosperm evolution by the duplication of the gene encoding deoxyhypusine synthase. To test whether high levels of homospermidine suffice for conversion, at least in traces, to precursors of pyrrolizidine alkaloids, transgenic tobacco plants were generated expressing homospermidine synthase. Analyses of the polyamine content revealed that, in the transgenic plants, about 80% of spermidine was replaced by homospermidine without any conspicuous modifications of the phenotype. Tracer feeding experiments and gas chromatographic analyses suggested that these high levels of homospermidine were not sufficient to explain the formation of alkaloid precursors. These results are discussed with respect to current models of pathway evolution. PMID- 19543983 TI - Metabolism of acenaphthylene via 1,2-dihydroxynaphthalene and catechol by Stenotrophomonas sp. RMSK. AB - Stenotrophomonas sp. RMSK capable of degrading acenaphthylene as a sole source of carbon and energy was isolated from coal sample. Metabolites produced were analyzed and characterized by TLC, HPLC and mass spectrometry. Identification of naphthalene-1,8-dicarboxylic acid, 1-naphthoic acid, 1,2-dihydroxynaphthalene, salicylate and detection of key enzymes namely 1,2-dihydroxynaphthalene dioxygenase, salicylaldehyde dehydrogenase and catechol-1,2-dioxygenase in the cell free extract suggest that acenaphthylene metabolized via 1,2 dihydroxynaphthalene, salicylate and catechol. The terminal metabolite, catechol was then metabolized by catechol-1,2-dioxygenase to cis,cis-muconic acid, ultimately forming TCA cycle intermediates. Based on these studies, the proposed metabolic pathway in strain RMSK is, acenaphthylene --> naphthalene-1,8 dicarboxylic acid --> 1-naphthoic acid --> 1,2-dihydroxynaphthalene --> salicylic acid --> catechol --> cis,cis-muconic acid. PMID- 19543984 TI - Myocardial electrical activity does not affect myocardial electrical impedance measurements. AB - OBJECTIVE: Myocardial electrical impedance (MEI) has shown to be an effective indicator of myocardial ischemia. We have previously developed a novel monitor for measuring MEI in near-real time. The object of this study was to test whether drug-induced changes in the frequency of the periodic myocardial electrical activity, as measured by the heart rate (HR), affect MEI measurements made with our monitor. METHODS: Thirty dogs were randomly assigned to one of three study groups (placebo, isoproterenol or esmolol) and then anesthetized with sodium thiamylal, intubated, ventilated, given isoflurane, and had venous, arterial, and pulmonary artery catheters placed. Median sternotomy was performed to facilitate myocardial exposure and to allow the left anterior descending (LAD) coronary artery to be isolated. Following baseline measurements, saline (control), isoproterenol or esmolol was administered and the LAD coronary artery was occluded in a timed sequence in order to study the effects of heart rate changes and demonstrate induced myocardial ischemia on MEI. RESULTS: Isoproterenol raised the HR and esmolol lowered the HR without affecting our MEI measurements. Myocardial electrical impedance increased during LAD coronary artery occlusion in all groups, as previously shown. CONCLUSION: These results demonstrate that our MEI monitor is unaffected by the frequency of the periodic myocardial electrical activity that generates the HR. PMID- 19543985 TI - Cancer profile of migrants from the Former Soviet Union in Germany: incidence and mortality. AB - This study compares cancer mortality and incidence of ethnic German migrants from the Former Soviet Union (FSU) in Germany. Data were obtained from two migrant cohorts residing in the federal state of North Rhine-Westphalia (NRW) (n = 34,393) and Saarland (n = 18,619). Vital status of the NRW cohort was ascertained through local population registries. Causes of death were obtained from the NRW statistical office or from local health offices. Cancer incidence of the Saarland cohort was derived from the Saarland cancer registry using record linkage. From 1990 to 2005, we observed 708 cancer deaths and 586 incident cancer cases. In males, both cancer incidence and cancer mortality were similar to the German population. Female cancer incidence and mortality were lower, the latter significantly. Site-specific standardized mortality and incidence ratios showed great variation in comparison to Germans and were remarkably similar to each other for most sites. Lung cancer was elevated among males, but lower among females. Stomach cancer was higher contrasting with lower ratios for prostate cancer, male colorectal cancer, and female breast cancer. Results confirm that FSU-migrants suffer from cancers, which may be prevented by prevention programs. Furthermore, we cannot conclude a different health-seeking behavior compared to Germans. PMID- 19543986 TI - Measuring cervical cancer risk: development and validation of the CARE Risky Sexual Behavior Index. AB - OBJECTIVES: To develop and validate a risky sexual behavior index specific to cervical cancer research. METHODS: Sexual behavior data on 428 women from the Community Awareness Resources and Education (CARE) study were utilized. A weighting scheme for eight risky sexual behaviors was generated and validated in creating the CARE Risky Sexual Behavior Index. Cutpoints were then identified to classify women as having a low, medium, or high level of risky sexual behavior. RESULTS: Index scores ranged from 0 to 35, with women considered to have a low level of risky sexual behavior if their score was less than six (31.3% of sample), a medium level if their score was 6-10 (30.6%), or a high level if their score was 11 or greater (38.1%). A strong association was observed between the created categories and having a previous abnormal Pap smear test (p < 0.001). CONCLUSIONS: The CARE Risky Sexual Behavior Index provides a tool for measuring risky sexual behavior level for cervical cancer research. Future studies are needed to validate this index in varied populations and test its use in the clinical setting. PMID- 19543988 TI - Heavy metals in Patella caerulea (Mollusca, Gastropoda) in polluted and non polluted areas from the Iskenderun Gulf (Mediterranean Turkey). AB - The concentrations of Cd, Cu, Zn, Fe, Pb, Ni, and Co were measured in gastropod mollusks Patella caerulea in the Mediterranean area. The organisms were collected at two coastal sites in Iskenderun Gulf during winter, spring, summer, and autumn 2008. Samples of the digestive gland, gill, and muscle were analyzed for heavy metals. The aim of study is to determine heavy metal levels in tissues of P. caerulea in different locations. Tissues of P. caerulea from the polluted site showed metal concentrations appreciably higher than unpolluted organisms. The highest metal levels were registered in the digestive gland of P. caerulea. Generally, digestive gland and gills showed higher metal concentrations than muscle. The average concentrations of heavy metals analyzed exhibited the following decreasing order: Fe>Zn>Cu>Ni>Cd>Pb>Co for both stations 1 and 2. Seasonal changes in metal concentrations were observed in the tissues of P. caerulea from a polluted and an unpolluted population. PMID- 19543987 TI - Access to mammography screening in a large urban population: a multi-level analysis. AB - OBJECTIVE: To understand area-based sociodemographics, physician and medical practice characteristics, and community indicators associated with mammography use in Los Angeles County. An earlier multi-level analysis by Gumpertz et al. found that distance to the nearest mammography facility helped explain the higher proportion of Latinas diagnosed with late stage breast cancer compared with non Latina Whites in Los Angeles County. Our study examined whether Latinas also have lower rates of mammography use. METHODS: We used a multi-level spatial modeling approach to examine individual and community level associations with mammography use among a diverse group of women aged 40-84 years in Los Angeles County. To build our multi-level spatial data set, we integrated five data sources: (1) 2001 California Health Interview Survey (CHIS) data, (2) 2001 Food and Drug Administration (FDA) certified mammography facility data, (3) 2003 LA Transit Authority data, (4) 2000 US Decennial Census data, and (5) 2001 Community Tracking Study (CTS) Physician's Survey data. RESULTS: Our study confirmed for Los Angeles County many associations for mammography use found in other locations. An unexpected finding was that women with limited English proficiency (predominantly Latina) were significantly more likely to have had a recent mammogram than English-proficient women. We also found that, after controlling for other factors, mammography use was higher in neighborhoods with a greater density of mammography facilities. CONCLUSION: Women with limited English proficiency were especially likely to report recent mammography in Los Angeles. This unexpected finding suggests that the intensive Spanish-language outreach program conducted by the Every Woman Counts (EWC) Program in low-income Latina communities in Los Angeles has been effective. Our study highlights the success of this targeted community-based outreach conducted between 1999 and 2001. These are the same populations that Gumpertz et al. identified as needing intervention. It would be useful to conduct another study of late-stage diagnosis in Los Angeles County to ascertain whether increased rates of mammography have also led to less late-stage diagnosis among Latinas in the neighborhoods where they are concentrated in Los Angeles. PMID- 19543991 TI - Retracted article: residues and dissipation dynamics of spirodiclofen in apple and soil. PMID- 19543989 TI - Trace metal dynamics in fishes from the southwest coast of India. AB - The concentration of Fe, Co, Ni, Cu, Zn, Cd, and Pb in the muscle tissue of marine fishes like Lates calcarifer, Nemipterus japonicus, Caranx melampygus, Rastrelliger kanagurta, and Cyanoglossus macrostomus was estimated from samples collected in the continental shelf waters off Kochi and Mangalore on southwest coast of India. Species-specific and spatially heterogeneous patterns of tissue metals loads were apparent within the pelagic and demersal fish species for the two regions. The concentration ranges of Fe (541.60 to 649.60 ppm), Ni (12.12 to 13.92 ppm), and Cu (3.09 to 3.62 ppm) were higher in the demersal species C. melampygus, whereas Co (9.10 to 11.80 ppm) and Zn (79.30 to 84.30 ppm) were higher in the pelagic species L. calcarifer and Cd (4.35 to 6.38 ppm) were higher in the demersal species N. japonicus, possibly due to enhanced bioavailability of these metals from ecological processes associated with upwelling during the summer monsoon. The fish species showed a great capacity to accumulate metals, with highest bioaccumulation for the essential element iron and lowest bioaccumulation for the non-essential element lead. Among the demersal species, C. melampygus and N. japonicus had high concentration factors for the metals Fe (280,268 to 322,808), Ni (88,252 to 96,891), Cu (2,351 to 2,600), and Cd (29,637 to 32,404). In contrast, the pelagic species L. calcarifer and R. kanagurta had high concentration factors for the metals Zn (40,812 to 46,892), Co (280,285 to 423,037), and Pb (854 to 1,404). PMID- 19543990 TI - Trace elements as tracers of environmental pollution in the canal sediments (alluvial formation of the Danube River, Serbia). AB - The purpose of this study was to examine the levels of trace elements and to discuss the origin and mobility of these contaminants in the canal sediments (alluvial formation of the Danube River). The most significant fractions binding all of the studied elements were oxides and silicates. The high proportion of elements in the residual fraction and the generally low contents of extractable elements reflected the background geochemical conditions. The contents of trace elements (except Hg and As) were dominantly controlled by the presence of Fe and Mn oxides as well as by the grain size and the geochemical composition of the sediment. The studied alluvial sediments were not heavily contaminated; there were only few sites where an anthropogenic influence on the concentrations of some metals (Cu, Cd, Zn, and Pb) was noticed. The results from this study show that impact of Pancevo industrial zone on the quality of the Danube River and its contamination with trace element is minor. PMID- 19543992 TI - TSP, PM depositions, and trace elements in the vicinity of a cement plant and their source apportionments using chemical mass balance model in Izmir, Turkey. AB - Total suspended particles mass concentrations (TSP) and bulk depositions of particulate matter (PM depositions) were measured around a cement plant located in the multi-impacted area to assess the affect of the plant on the ambient air in the vicinity in Izmir, Turkey. TSP samples were collected five times a month whereas PM depositions were sampled monthly at four sites between August 2003 and January 2004. The concentrations of Al, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, Pb, V, and Zn in TSP and PM depositions (except Cu) were reported. Chemical mass balance (CMB) receptor model with local source profiles was run in order to calculate the source contributions of the PM sources to the concentrations of TSP, PM depositions, and trace elements. Traffic was found to be the major contributor to TSP whereas PM depositions dominantly result from area sources including several stone quarries, concrete plants, lime kilns, and asphalt plants in the region. CMB model results indicate that the cement plant is a significant contributor to TSP, PM depositions, and trace elements, particularly Cd. PMID- 19543993 TI - Utilization of the water quality index method as a classification tool. AB - The study comprised modification of the Canadian Council of Ministers of the Environment (CCME) Water Quality Index (CCMEWQI) to obtain a tool in classification of surface waters according to quality defined by the European Legislation-75/440/EEC. Three categories were proposed, and the category ranges of CCMEWQI have been modified depending on the objective chosen. The application of the CCMEWQI with modified categorization scheme was demonstrated to assess overall water quality by integrating observed water quality determinants in the Kucuk Menderes Basin, Turkey. In this scope, the samples analyzed for pH, total dissolved solids (TDS), chlorides (Cl), nitrate-nitrogen (NO3-N), dissolved oxygen (DO), biochemical oxygen demand (BOD5), sulfate (SO4), and boron (B), variables taken monthly over 2 years from the five monitoring sites, were processed. Results revealed that the overall surface water mainly fell within the A2 water class. The CCMEWQI with modified categorization scheme is believed to assist water managers to integrate and interpret the picture of overall water quality based on the European legislation concerning the quality required of surface water intended for the abstraction of drinking water in the Member States (75/440/EEC). PMID- 19543995 TI - Assessment of ground water quality for drinking purpose, District Nainital, Uttarakhand, India. AB - The ground water quality of District Nainital (Uttarakhand, India) has been assessed to see the suitability of ground water for drinking and irrigation applications. This is a two-part series paper and this paper examines the suitability of ground water including spring water for drinking purposes. Forty ground water samples (including 28 spring samples) were collected during pre- and post-monsoon seasons and analyzed for various water quality constituents. The hydrochemical and bacteriological data was analyzed with reference to BIS and WHO standards and their hydrochemical facies were determined. The concentration of total dissolved solids exceeds the desirable limit of 500 mg/L in about 10% of the samples, alkalinity values exceed the desirable limit of 200 mg/L in about 30% of the samples, and total hardness values exceed the desirable limit of 300 mg/L in 15% of the samples. However, no sample crosses the maximum permissible limit for TDS, alkalinity, hardness, calcium, magnesium, chloride, sulfate, nitrate, and fluoride. The concentration of chloride, sulfate, nitrate, and fluoride are well within the desirable limit at all the locations. The bacteriological analysis of the samples does not show any sign of bacterial contamination in hand pump and tube-well water samples. However, in the case of spring water samples, six samples exceed the permissible limit of ten coliforms per 100 ml of sample. It is recommended that water drawn from such sources should be properly disinfected before being used for drinking and other domestic applications. Among the metal ions, the concentration of iron and lead exceeds the permissible limit at one location whereas the concentration of nickel exceeds the permissible limit in 60 and 32.5% of the samples during pre- and post-monsoon seasons, respectively. The grouping of samples according to their hydrochemical facies indicates that majority of the samples fall in Ca-Mg-HCO(3) hydrochemical facies. PMID- 19543994 TI - Transfer of natural and anthropogenic radionuclides to ants, bryophytes and lichen in a semi-natural ecosystem. AB - Few data are available to quantify the transfer of both natural and anthropogenic radionuclides to detritivorous invertebrates to facilitate estimation of the internal dose to such biota in models used to assess radiation exposure. To enhance the available data, activity concentrations of (137)Cs, (40)K, (90)Sr, (239 + 240)Pu, (241)Am, (235)U and (238)U were measured in ants (Formicidae) and corresponding undisturbed soil collected from the Zlatibor mountain in Serbia and ant/soil concentration ratios (CR) calculated. The (241)Am concentration ratios for ants were fourfold higher than those calculated for ants in a previous study whereas they are similar to the more numerous data previously reported for a range of detritivorous invertebrates in other studies. CR values for (137)Cs in ants were similar to the few other reported values and slightly lower than those for a range of detritivorous invertebrates. Those for (239 + 240)Pu were slightly higher than those for ants in two other studies but they were close to upper limit of a range of data reported for detritivorous invertebrates. All the CR values will be included in a future revision of the ERICA Tool database and will particularly improve the information available for uranium. PMID- 19543996 TI - The Rengen Grassland experiment: bryophytes biomass and element concentrations after 65 years of fertilizer application. AB - The Rengen Grassland Experiment in Germany, established in 1941, consists of the following fertilizer treatments applied under a two cut management: control, Ca, CaN, CaNP, CaNP-KCl, and CaNP-K(2)SO(4). The aim of this study was (1) to identify effects of fertilizer application on biomass and species composition of bryophytes and (2) to investigate the impact of fertilizer application on macro- (N, P, K, Ca, Mg), micro- (Cu, Fe, Mn, Zn), and toxic (As, Cd, Cr, Pb, Ni) element concentrations in bryophyte biomass. In June 2006, Rhytidiadelphus squarrosus was the only bryophyte species recorded in the control. In treatment Ca, R. squarrosus was the dominant bryophyte species whereas Brachythecium rutabulum occurred sporadically only in a single plot of that treatment. The latter was the only bryophyte species collected in CaN, CaNP, CaNP-KCl, and CaNP K(2)SO(4) treatments. Dry matter accumulation of bryophytes was highest in the control (180 g m(-2)) followed by Ca (46 g m(-2)), CaNP (25 g m(-2)), CaNP-KCl (15 g m(-2)), CaNP-K(2)SO(4) (9 g m(-2)), and CaN (2 g m(-2)) treatments. A negative correlation between biomass production of bryophytes and dry matter production of vascular plants was revealed up to a threshold value of 400 g m( 2). Above this limit, biomass production of bryophytes remained obviously unaffected by further increase in biomass production of vascular plants. A significant effect of treatment on As, Cd, Cr, Fe, Mn, Ni, Pb, P, Ca, Mg, K, and N concentrations was revealed. Concentrations of these elements were a function of amount of elements supplied with fertilizers. Bryophytes seem to be promising bio-indicators not only for airborne deposition of toxic element but also for fertilizer introduced as well. PMID- 19543997 TI - Aluminium concentrations in Swedish forest streams and co-variations with catchment characteristics. AB - The negative effects of elevated concentrations of inorganic aluminium on aquatic organisms are well documented. Acid deposition is often cited as a main driver behind the mobilisation and speciation of aluminium in soils and surface waters. In the study, we tested the hypothesis that sulphur deposition is the main driver for elevated concentrations of inorganic aluminium in 114 base poor, boreal Swedish streams. However, the deposition of anthropogenic sulphate has decreased substantially since it peaked in the 1970s, and at the current deposition levels, we hypothesise that local site parameters play an important role in determining vulnerability to elevated concentrations of inorganic aluminium in boreal stream waters. Presented here are the results of a principal components analysis of stream water chemistry, acid deposition data and local site variables, including forest composition and stem volume. It is shown that the concentrations of both organic and inorganic aluminium are not explained by either historical or current acid deposition, but are instead explained by a combination of local site characteristics. Sites with elevated concentrations of inorganic aluminium were characterised by small catchments (<500 ha) dominated by mature stands of Norway spruce with high stem volume. Using data from the Swedish National Forest Inventory the area of productive forest land in Sweden with a higher vulnerability for elevated inorganic aluminium concentrations in forests streams is approximately 1.5 million hectares or 7% of the total productive forest area; this is higher in the south of Sweden (10%) and lower in the north (2%). A better understanding of the effects of natural processes and forest management in controlling aquatic inorganic aluminium concentrations is therefore important in future discussions about measures against surface water acidification. PMID- 19543998 TI - Characterization of dissolved organic carbon at low levels in environmental waters by microfluidic-chip-based capillary gel electrophoresis with a laser induced fluorescence detector. AB - A microfluidic analytical system for characterization of dissolved organic carbon (DOC) in environmental waters, based on a capillary gel electrophoresis (CGE) device with a laser-induced fluorescence (LEF) detector, was developed. The applied voltage and the running buffer were investigated to control the simple floating injection and CGE separation for convenient cross-type microchips made from polymethyl-methacylate. We obtained reproducible peaks for standard organic solutions and the determination time is less than 70 s. The values of the relative standard deviation (RSD) were 0.17-2.01% for repetitive injection (n = 12). We demonstrated high-throughput characterization of DOC in environmental water from the Biwa Lake and the Hino River using microfluidic chip and determined that the content of DOC in the Biwa Lake changed with the seasons. PMID- 19543999 TI - Application of ANN and ANFIS models for reconstructing missing flow data. AB - Hydrological yearbooks, especially in developing countries, are full of gaps in flow data series. Filling missing records is needed to make feasibility studies, potential assessment, and real-time decision making. In this research project, it was tried to predict the missing data of gauging stations using data from neighboring sites and a relevant architecture of artificial neural networks (ANN) as well as adaptive neuro-fuzzy inference system (ANFIS). To be able to evaluate the results produced by these new techniques, two traditionally used methods including the normal ratio method and the correlation method were also employed. According to the results, although in some cases all four methods presented acceptable predictions, the ANFIS technique presented a superior ability to predict missing flow data especially in arid land stations with variable and heterogeneous data. Comparing the results, ANN was also found as an efficient method to predict the missing data in comparison to the traditional approaches. PMID- 19544000 TI - Relationships among functional markers, management, and husbandry in sheep: a Mediterranean case study. AB - Most sheep farmers are aware of the importance of monitoring animal health and well-being for profitable sheep production. Unfortunately, there are only a few benchmarked functional measures of sheep well-being but much can be gained from our understanding of other species. Moreover, comprehensive monitoring programs may be complex and relatively expensive to implement. Hence, this work reports the results of a research study on the usefulness of functional markers in measuring dairy sheep well-being, taking into account farm management and environmental conditions. The study was conducted on 11 farms breeding Italian islander sheep breeds. The husbandry and management parameters of each farm were assessed and, based on the findings, the farms were scored in ascending quality order. Flock information concerned housing, milking system, pen size, grazing hours, health management, and stockmanship. Medical history, clinical data, the most relevant haematological, chemical and biochemical parameters, as well as the haemoglobin genotype were recorded for 415 individuals. The whole data-set was analyzed by Spearman correlation and multivariate statistical procedures, showing that albumin, serum alkaline phosphatase, haematocrit, and haemoglobin were the most significant functional markers of a flock's general conditions. Haematocrit and haemoglobin reflect animal health status, while albumin and serum alkaline phosphatase are a measure of nutritional status and physical activity, respectively. These are objective parameters, which can be easily measured from blood samples and have proved to be effective for grouping to interpret animal well-being. PMID- 19544001 TI - Effect of mitratapide on body composition, body measurements and glucose tolerance in obese Beagles. AB - The objective of this study was to confirm that weight loss after treatment with mitratapide (Yarvitan(r)) is loss of adipose tissue. Obese dogs were treated with the recommended treatment schedule of mitratapide. Dual-energy X-ray absorptiometry (DEXA) was done before and after the treatment schedule. Body weight, feed consumption and pelvic circumference were recorded and a glucose tolerance test was performed. Dualenergy X-ray absorptiometry measurements showed an impressive loss of fat tissue, corresponding to a mean loss of approximately 41.6% of the body fat mass recorded before treatment. After treatment with mitratapide, the mean body fat percentage had returned within the normal range. At the end of the study, the dogs had lost on average 14.2% of their body weight and 15.2% of their pelvic circumference compared to baseline. The results also suggest that losing weight with mitratapide might help to reverse insulin resistance. PMID- 19544003 TI - Within-country variation in the ability of ruminants to degrade DHP following the ingestion of Leucaena leucocephala--a Thailand experience. AB - Goats fed Leucaena leucocephala (leucaena) at an experimental site in Thailand were shown to be excreting DHP in their urine. This was unexpected as earlier results from another site had shown that goats and cattle fed leucaena did not excrete DHP and so possessed DHP-degrading bacteria. Goats sampled near the earlier sample site excreted no DHP in their urine. Rumen fluid taken from these goats was successfully used to transfer DHP--degrading ability to the goats at the Experimental site some 350 km away that did not show the presence of DHP degrading bacteria. Degradation of mimosine in-vitro and excretion of DHP in the urine ceased 72 hr after addition of rumen fluid and infusion with rumen fluid from protected goats, respectively. The situation in Thailand may not be unique. Countries where leucaena is fed should check that animals are protected. Fortunately, the ferric chloride urine test is simple to use and effective in detecting the problem and also the recovery after transfer of rumen fluid from protected animals. PMID- 19544004 TI - Traditional llama husbandry and breeding management in the Ayopaya region, Bolivia. AB - The llama claims the largest population of the domestic South American camelids, most of which are raised in Bolivia. More than 53,000 rural families are dedicated to llama husbandry as part of their livelihood strategy. Contemporary Andean societies deliberately select animals for specific traits and employ substantial livestock management to secure subsistence. This study presents traditional llama husbandry and breeding management activities in the Ayopaya region, Bolivia. Traditional selection traits for male and female llamas are documented and assessed by a ranking and a ratio-scaled evaluation. Husbandry and management parameters are in concordance with other studies conducted in the region, but show a high variation. Average llama herd sizes are rather small (mu = 45.6). In some herds, breeding males are utilized for a long time and mix with other herds, causing concerns about inbreeding. Preferred trait groups for llama males according to farmers' responses were body conformation, fibre, testicle conformation, fleece colour and height at withers. Traditional selection criteria generally relate to the phenotype, but also include the commercially interesting fibre trait. The presented results should be considered in breeding and management programmes for the respective llama population to ensure sustainable use of this genetically and culturally valuable resource. PMID- 19544002 TI - Spermidine levels are implicated in heavy metal tolerance in a spermidine synthase overexpressing transgenic European pear by exerting antioxidant activities. AB - To verify whether spermidine synthase (SPDS) can confer long-term multi-heavy metal tolerance, in vitro shoots of a transgenic European pear (Pyrus communis L. 'Ballad') line #32 overexpressing apple SPDS (MdSPDS1), as well as a wild type (WT) line, were subjected to stress using either CdCl(2), PbCl(2), ZnCl(2), or a combination thereof. Based on either shoot height increment or fresh weight and morphological changes upon heavy metal stress, the performance of the transgenic line #32 was better than that of WT. Although SPDS expression levels and spermidine (Spd) contents in line #32 were higher than those in WT, possibly due to transgene (MdSPDS1) expression, no obvious inductions of SPDS expression and increases in Spd-content were observed by long-term stress treatments in both lines. When the glutathione (GSH) content was compared with or without stress in each line, GSH was significantly depleted in line #32 with stress, but not as much as in WT. The activities of glutathione reductase and superoxide dismutase and the content of malondialdehyde, an indicator for lipid peroxidation, changed upon stress toward a more favorable status for survival in line #32 than in WT. These antioxidant parameters were positively related to Spd-content. The accumulation of heavy metals tended to be less in line #32 than in WT except for Zn stress, and the Ca content showed an opposite trend. These results suggest that Spd-levels are implicated in enhanced heavy metal tolerance, possibly by exerting an antioxidant activity as well as by the properties of Spd per se including metal chelator. PMID- 19544005 TI - Temporal changes in endogenous estrogens and expression of behaviors associated with estrus during the periovulaory period in Murrah buffaloes (Bubalus bubalis). AB - The objective of this study were (1) to establish the duration of behavioral estrus signs and timing of ovulation in Murrah buffaloes (n = 10) and (2) to determine relationship between behavioral estrus signs with change in plasma estrogen concentrations. Estrus and its behavioral signs were detected at hourly intervals by visual observations, per recta examination of genitalia and bull parading four times in a day for 30 minutes each. Among the behavioral signs of estrus, swollen vulva (80%) was the best indicator of estrus followed by excitement (70%). Among the duration of behavioral estrus signs the first and longest duration of estrus signs was swollen vulva which was seen upto 19.8 +/- 0.8 h after onset of estrus. The mean total duration of estrus symptoms from appearance to disappearance of all the behavioral estrus symptoms was 23.5 +/- 1.7 h. All the behavioral estrus symptoms were observed during the period of estrogen surge. Endocrine profile during the periestrus period showed that the mean peak concentrations of total estrogen 23.9 +/- 3.9 pg/ml occurred at 8.8 +/- 1.7 h after onset of estrus. The average number of estrus symptoms observed per animal during onset of spontaneous estrus was 5.7. Ovulation occurred after 37.4 +/- 1.7 h after onset of estrus and 13.4 +/- 1.0 h after end of total estrogen surge respectively. In conclusion, our results suggest that all signs of behavioral estrus occurred during the preovulatory rise in estrogens. The first sign of estrus to be observed was a swollen vulva and this symptom persisted the longest. PMID- 19544006 TI - Effects of the level of sugarcane molasses on growth and carcass performance of Caribbean growing pigs reared under a ground sugarcane stalks feeding system. AB - The influence of the level of sugarcane (SC) molasses on growth performance, carcass traits, and meat quality in Creole (CR) growing pigs fed with ground sugarcane stalks (GCS)-based diet was studied in a mixed farming system context. The aim of the study was to optimize the growth performance of CR pigs with SC molasses as an energy source in this unconventional feeding. A total of 32 CR pigs were used from 30 to 60 kg of body weight (BW). The experimental dietary treatments consisted of four levels of inclusion of SC-molasses (200, 400, 600, and 800 g DM/d/pig) into a GCS diet, for diets 1, 2, 3 and 4 respectively. The GCS allowance was based on live BW (170 g/kg BW/d) and the diets were supplemented with a soya-bean meal supplement (350 g/d of a 49.2% CP and 16.6 MJ DE/kg). All the pigs were slaughtered at 60 kg BW. Increasing the level of molasses did not affect (p > 0.05) average BW gain (254 g/d), CP intake (154 g/d) and sugar extraction rate from the total ration (85%). A gradual inclusion of molasses in a GCS-based diet did not affect the carcass and meat quality of CR pigs. In conclusion, molasses supplementation does not allow the increase of growth performance in GCS fed pigs. PMID- 19544007 TI - Deriving room temperature excitation spectra for photosystem I and photosystem II fluorescence in intact leaves from the dependence of FV/FM on excitation wavelength. AB - The F(0) and F(M) level fluorescence from a wild-type barley, a Chl b-less mutant barley, and a maize leaf was determined from 430 to 685 nm at 10 nm intervals using pulse amplitude-modulated (PAM) fluorimetry. Variable wavelengths of the pulsed excitation light were achieved by passing the broadband emission of a Xe flash lamp through a birefringent tunable optical filter. For the three leaf types, spectra of F(V)/F(M) (=(F(M) - F(0))/F (M)) have been derived: within each of the three spectra of F(V)/F(M), statistically meaningful variations were detected. Also, at distinct wavelength regions, the (V)/F(M) differed significantly between leaf types. From spectra of F(V)/F (M), excitation spectra of PS I and PS II fluorescence were calculated using a model that considers PS I fluorescence to be constant but variable PS II fluorescence. The photosystem spectra suggest that LHC II absorption results in high values of F(V)/F(M) between 470 and 490 nm in the two wild-type leaves but the absence of LHC II in the Chl b-less mutant barley leaf decreases the F(V)/F(M) at these wavelengths. All three leaves exhibited low values of F(V)/F(M) around 520 nm which was tentatively ascribed to light absorption by PS I-associated carotenoids. In the 550-650 nm region, the F(V)/F(M) in the maize leaf was lower than in the barley wild-type leaf which is explained with higher light absorption by PS I in maize, which is a NADP-ME C(4) species, than in barley, a C(3) species. Finally, low values of F(V)/F(M) at 685 in maize leaf and in the Chl b-less mutant barley leaf are in agreement with preferential PS I absorption at this wavelength. The potential use of spectra of the F(V)/F(M) ratio to derive information on spectral absorption properties of PS I and PS II is discussed. PMID- 19544008 TI - Low-temperature single-molecule spectroscopy on photosynthetic pigment-protein complexes from purple bacteria. AB - The primary reactions of purple bacterial photosynthesis take place within two well characterized pigment-protein complexes, the core Reaction Center-Light Harvesting 1 (RC-LH1) complex and the more peripheral Light Harvesting 2 (LH2) complex. These antenna complexes serve to absorb incident solar radiation and to transfer it to the reaction-centers, where it is used to 'power' the photosynthetic redox reaction. This review provides an overview of how the character of the electronically excited states of these pigment-protein complexes are determined by quantum mechanics and how the respective spectral signatures can be observed by single-molecule spectroscopy. PMID- 19544009 TI - An unusual genetic variant in the MOCS1 gene leads to complete missplicing of an alternatively spliced exon in a patient with molybdenum cofactor deficiency. AB - Molybdenum cofactor deficiency (MOCOD) is a rare inherited metabolic disorder resulting in the combined deficiency of aldehyde oxidase (AO, EC 1.2.3.1), xanthine dehydrogenase (XDH, EC 1.1.1.204), and sulfite oxidase (SUOX, EC 1.8.3.1). The majority of patients typically present soon after birth with intractable seizures, developmental delay and lens dislocation and do not survive early childhood. Milder cases have been reported. We report an unusual mutation in the MOCS1 gene associated with a relatively mild clinical phenotype, in a patient who presented with normal uric acid (UA) levels in plasma. We also report a new MOCS1 mRNA splice variant in the 5' region of the gene. MOCS1 genomic DNA and cDNA from peripheral blood leukocytes were sequenced. MOCS1 mRNA splice variants were amplified with fluorescently labelled primers and quantitated. A novel homozygous mutation MOCS1c.1165+6T > C in intron 9 resulting in miss splicing of exon 9 was found. Multiple alternatively spliced MOCS1 transcripts have been previously reported. A new MOCS1 transcript in the 5' - exon 1 region was identified in both patient and controls. This new transcript derived from the Larin variant and lacked exon 1 d. PMID- 19544011 TI - What is at stake in the predictive genetic testing of children? PMID- 19544010 TI - Enhancement of Candida albicans virulence after exposition to cigarette mainstream smoke. AB - The habit of cigarette smoking is associated with higher oral candidal carriage and possible predisposition to oral candidosis. The effects of exposure to smoke on the virulence properties of oral yeasts remain obscure. Hence, we showed in vitro the effect of cigarette smoke condensate (CSC) on ten clinical isolates of Candida albicans obtained from nonsmoking volunteers, as well the type-strain CBS562. CSC was generated by complete burn of five commercial cigarettes in an in house smoking machine and used to prepare the culture broth in which the strains were grown. In 24-h intervals (T(24), T(48), and T(72)), the cells were harvested, washed, subcultured, and the resultant growth were evaluated for possible variations for secreted aspartyl protease, phospholipase, chondroitinase, and hemolysins, adhesion to acrylic and cell surface hydrophobicity (CSH). The results indicated a temporal increase in the secretion rates of enzymes, particularly when yeast cells were exposed to CSC for 48-72 h (P < 0.05). Similarly, adhesion to acrylic and CSH increased with exposure period (P < 0.05). Based on foregoing, we concluded that CSC may promote significant enhance in the secretion of candidal histolytic enzymes and adherence to denture surfaces, thereby promoting oral yeast carriage and possible infection. PMID- 19544012 TI - MicroRNAs in mental health: from biological underpinnings to potential therapies. AB - Psychiatric illnesses are disabling disorders with poorly understood underlying pathophysiologies. However, it is becoming increasingly evident that these illnesses result from disruptions across whole cellular networks rather than any particular monoamine system. Recent evidence continues to support the hypothesis that these illnesses arise from impairments in cellular plasticity cascades, which lead to aberrant information processing in the circuits that regulate mood, cognition, and neurovegetative functions (sleep, appetite, energy, etc.). As a result, many have begun to consider future therapies that would be capable of affecting global changes in cellular plasticity to restore appropriate synaptic function and neuronal connectivity. MicroRNAs (miRNAs) are non-coding RNAs that can repress the gene translation of hundreds of their targets and are therefore well-positioned to target a multitude of cellular mechanisms. Here, we review some properties of miRNAs and show they are altered by stress, glucocorticoids, mood stabilizers, and in a particular psychiatric disorder, schizophrenia. While this field is still in its infancy, we consider their potential for regulating behavioral phenotypes and targeting key predicted signaling cascades that are implicated in psychiatric illness. Clearly, considerable research is required to better determine any therapeutic potential of targeting miRNAs; however, these agents may provide the next generation of effective therapies for psychiatric illnesses. PMID- 19544013 TI - Possible association of prokineticin 2 receptor gene (PROKR2) with mood disorders in the Japanese population. AB - Several investigations have suggested that disruption of circadian rhythms may provide the foundation for the development of mood disorders such as bipolar disorder (BP) and major depressive disorder (MDD). Recent animal studies reported that prokineticin 2 or prokineticin 2 receptor gene deficient mice showed disruptions in circadian and homeostatic regulation of sleep. This evidence indicates that prokineticin 2 gene (PROK2) and prokineticin 2 receptor gene (PROKR2) are good candidate genes for the pathogenesis of mood disorders. To evaluate the association between PROK2, PROKR2, and mood disorders, we conducted a case-control study of Japanese samples (151 bipolar patients, 319 major depressive disorder patients, and 340 controls) with four and five tagging SNPs in PROK2 or PROKR2, respectively, selected by HapMap database. We detected a significant association between PROKR2 and major depressive disorder and bipolar disorder in the Japanese population. In conclusion, our findings suggest that PROKR2 may play a role in the pathophysiology of mood disorders in the Japanese population. However, because our samples were small, it will be important to replicate and confirm these findings in other independent studies using larger samples. PMID- 19544014 TI - Introduction to proteomics. AB - In this chapter, the evolvement of proteomics from classical protein chemistry is depicted. The challenges of complexity and dynamics led to several new approaches and to the firm belief that a valuable proteomics technique has to be quantitative. Protein-based vs. peptide-based techniques, gel-based vs. non-gel based proteomics, targeted vs. general proteomics, isotopic labeling vs. label free techniques, and the importance of informatics are summarized and compared. A short outlook into the near future is given at the end of the chapter. PMID- 19544015 TI - High-resolution two-dimensional electrophoresis. AB - Two-dimensional gel electrophoresis (2-DE) with immobilized pH gradients (IPGs) combined with protein identification by mass spectrometry is currently the workhorse for the majority of ongoing proteome projects. Although alternative/complementary technologies, such as MudPIT, ICAT, or protein arrays, have emerged recently, there is up to now no technology that matches 2-DE in its ability for routine parallel expression profiling of large sets of complex protein mixtures. 2-DE delivers a map of intact proteins, which reflects changes in protein expression level, isoforms, or post-translational modifications. High resolution 2-DE can resolve up to 5,000 proteins simultaneously ( approximately 2,000 proteins routinely), and detect and quantify <1 ng of protein per spot. Today's 2-DE technology with IPGs has largely overcome the former limitations of carrier ampholyte-based 2-DE with respect to reproducibility, handling, resolution, and separation of very acidic or basic proteins. Current research to further advance 2-DE technology has focused on improved solubilization/separation of hydrophobic proteins, display of low abundance proteins, and reliable protein quantitation by fluorescent dye technologies. Here, we provide a comprehensive protocol of the current high-resolution 2-DE technology with IPGs for proteome analysis and describe in detail the individual steps of this technique, i.e., sample preparation and protein solubilization, isoelectric focusing in IPG strips, IPG strip equilibration, and casting and running of multiple SDS gels. Last but not the least, a section on how to circumvent the major pitfalls is included. PMID- 19544016 TI - Non-classical 2-D electrophoresis. AB - Classical 2-D electrophoresis (IEF/SDS 2-DE) using isoelectric focusing (IEF) and SDS-PAGE for the second dimension offers very high resolution for the separation of complex protein mixtures, but hydrophobic proteins can aggregate and are considerably under-represented in these 2-D gels. Non-classical 2-DE, as described here, summarizes several heterogeneous techniques, some of which, like BAC/SDS 2-DE and doubled SDS-polyacrylamide gel electrophoresis (dSDS-PAGE), intend to isolate the difficult hydrophobic proteins that are not accessible by classical 2-DE. Other types of non-classical 2-DE start with 1-D separation of native proteins and complexes, like blue-native electrophoresis (BNE), clear native electrophoresis (CNE), and high-resolution clear-native electrophoresis (hrCNE). These electrophoretic techniques can substitute for chromatographic isolation of protein complexes, and can even isolate supramolecular physiological assemblies. Subsequent resolution in second dimension can be denaturing to resolve the subunits of complexes, as exemplified with BNE/SDS 2-DE, or native like in BNE/BNE 2-DE (the latter using different cathode buffers for 1-D BNE and 2-D BNE). After isolation of highly pure membrane protein complexes by two native electrophoretic separations, the separation protocol may be finished by denaturing 2-DE like BAC/SDS or doubled SDS-PAGE. Thus, a four-dimensional electrophoretic system with minimal loss of protein results that is useful as an efficient micro-scale protein separation protocol, e.g. for mass spectrometric analyses. PMID- 19544017 TI - Protein detection and quantitation technologies for gel-based proteome analysis. AB - Numerous protein detection and quantitation methods for gel-based proteomics have been devised that can be classified in three major categories: (1) Universal (or "general") detection techniques, which include staining with anionic dyes (e.g., Coomassie brilliant blue), reverse (or "negative") staining with metal cations (e.g., imidazole-zinc), silver staining, fluorescent staining or labeling, and radiolabeling, (2) specific staining methods for the detection of post translational modifications (e.g., glycosylation or phosphorylation), and (3) differential display techniques for the separation of multiple, covalently tagged samples in a single two-dimensional electrophoresis (2-DE) gel, followed by consecutive and independent visualization of these proteins to minimize methodical variations in spot positions and in protein abundance, to simplify image analysis, as well as to improve protein quantitation by including an internal standard. The most important properties of protein detection methods applied in proteome analysis include high sensitivity (i.e., low detection limit), wide linear dynamic range for quantitative accuracy, reproducibility, cost-efficiency, ease of use, and compatibility with downstream protein identification or characterization technologies, such as mass spectrometry (MS). Regrettably, no single detection method meets all these requirements, albeit fluorescence-based technologies are currently favored for most applications; hence, the major focus of this chapter is on fluorescent-dye-based protein detection and quantitation techniques. Although satisfying results with respect to sensitivity and reproducibility are also obtained by methods based on radioactive labeling of proteins (which is still unsurpassed in terms of sensitivity), radiolabeling is, however, largely impractical for routine proteomic profiling because of the costs and the health and safety concerns associated with handling radioactive compounds. PMID- 19544018 TI - MALDI MS. AB - Matrix-assisted laser desorption/ionization (MALDI) is a key technique in mass spectrometry (MS)-based proteomics. MALDI MS is extremely sensitive, easy-to apply, and relatively tolerant to contaminants. Its high-speed data acquisition and large-scale, off-line sample preparation has made it once again the focus for high-throughput proteomic analyses. These and other unique properties of MALDI offer new possibilities in applications such as rapid molecular profiling and imaging by MS. Proteomics and its employment in Systems Biology and other areas that require sensitive and high-throughput bioanalytical techniques greatly depend on these methodologies. This chapter provides a basic introduction to the MALDI methodology and its general application in proteomic research. It describes the basic MALDI sample preparation steps and two easy-to-follow examples for protein identification including extensive notes on these topics with practical tips that are often not available in the Subheadings 2 and 3 of research articles. PMID- 19544019 TI - Capillary electrophoresis coupled to mass spectrometry for proteomic profiling of human urine and biomarker discovery. AB - Currently, the main focus of clinical proteome analysis is on detection and identification of polypeptides that significantly change owing to pathological changes. Capillary electrophoresis coupled online to an electrospray ionization time of flight mass spectrometer (CE-MS) allows the differential display of a large number of polypeptides in a single, reproducible, and time-limited step and enables the comparison of different protein profiles for biomarker discovery. In addition to the reproducibility of the CE-MS setup, many further steps including data processing and mining, usage of biomarkers for diagnosis, and biomarker sequencing are necessary to answer the demands of biomarker discovery of clinical significance. In this chapter, we discuss materials and methods for CE-MS-based clinical proteomics allowing the reproducible profiling of urine. PMID- 19544020 TI - A newcomer's guide to nano-liquid-chromatography of peptides. AB - LC-MS/MS is one of the most powerful techniques in the field of proteomics allowing high throughput identification of proteins out of complex protein mixtures. Besides high sample throughput, the analytical sensitivity is one of the major benefits of this technology. A prerequisite for sensitive LC-MS/MS approaches is chromatography with very low flow rates in the nanoliter per minute range, usually referred to as nano-liquid chromatography (nano-LC). However, to perform this separation technology, an appropriate instrumental setup as well experienced operators are a prerequisite. The aim of this chapter is to help nano LC newcomers to get introduced to this fascinating technology. Technical components of nano-LC systems like solvent delivery systems, sample injection systems, and nano-chromatography columns are described. Detailed procedures to mount, test, and operate the system are outlined, and advices for an effective troubleshooting are provided. PMID- 19544021 TI - Multidimensional protein identification technology. AB - Over the past years, large-scale analysis of proteomes gained increased interest to obtain a fast but nevertheless comprehensive overview about cellular protein content. While a complete proteome cannot be covered using current technologies because of its enormous diversity, subfractionation to reduce the complexity has become mandatory. While 2D-PAGE is well established as a high-resolution protein separation technique, it suffers from drawbacks, which can be overcome by using peptide separation methods based on multidimensional liquid chromatography. One of these technologies is multidimensional protein identification technology (MudPIT). It consists of two orthogonal separation systems--strong cation exchange (SCX) and reversed phase (RP)--coupled online in an automated fashion to mass spectrometric detection. This method offers the possibility to analyze high complex peptide mixtures in a single experiment. PMID- 19544022 TI - Characterization of platelet proteins using peptide centric proteomics. AB - In modern proteomics, undersampling of low abundant, cumbersome, and hydrophobic proteins states one of the major problems. To overcome this, especially in two 2D PAGE (two-dimensional polyacrylamide gel electrophoresis) eminent drawbacks, the so-called peptide-centric techniques have been developed. These approaches do not separate proteins prior to digestion, but instead proteolytically generate peptide mixtures after it. However, by this procedure already complex protein mixtures become even more extensive peptide mixtures. Particularly, when dealing with large proteomes, the generated sample complexity is vast and therefore difficult to analyze. When separated and analyzed by LC/MS, too many peptides may enter the mass spectrometer at a certain time point, and only a small fraction of ions is selected for subsequent MS/MS analysis. Although protein hydrophobicity and size play minor roles (as long as protease cleavage sites are accessible), low copy number can severely limit identification rates. To reduce the amount of peptides entering the mass spectrometer simultaneously without the loss of overall proteomic information, different techniques have been developed. Among these, an approach is represented by COFRADIC (Combined Fractional Diagonal Chromatography). COFRADIC is a chromatography-based technique enabling the sorting of peptides due to retention time shifts after a specific modification step. In the original approach, a complex peptide mixture is separated by a primary RP-HPLC (reversed-phase high-performance liquid chromatography) run and fractions are retained. Subsequently, these fractions are modified to specifically change retention times of peptides and separated in one or more secondary RP-HPLC runs. In this chapter, COFRADIC approaches for methionine or cysteine containing as well as N-terminal peptides are described. Besides the reduction of sample complexity, the major advantage of COFRADIC might be seen in its versatility. Nearly every feature unique for a subset of peptides, which can be specifically modified by a sorting reaction, is accessible for COFRADIC. Among these are protein phosphorylation, N-glycosylation, and in vivo protein processing sites. Finally, COFRADIC allows the analysis of large numbers of samples and is highly automatable. PMID- 19544023 TI - Identification of the molecular composition of the 20S proteasome of mouse intestine by high-resolution mass spectrometric proteome analysis. AB - In the last years, intracellular protein degradation by the proteasome has become a focus area of scientific interest. Here, we describe a proteomics approach for the molecular mapping of the constituents of the proteolytically active core particle, the constitutive 20S proteasome from mouse intestine. In addition to the proteomics workflow widely used for protein isolation, gel electrophoretic separation, in-gel digestion, and UV-MALDI mass spectrometry, high-resolution Fourier transform ion cyclotron resonance mass spectrometry using infrared-MALDI ionisation (IR-MALDI FTICR-MS) has been employed as an efficient method for protein identification by peptide mass fingerprint. The 20S proteasome subunits alpha1-alpha7 and beta1-beta7 were completely and unambiguously identified. In addition to subunits beta1 and beta2, the corresponding inducible subunits being part of the immuno-proteasome were identified. The subunit beta5i was found to completely replace the corresponding constitutive subunit, suggesting a high proteolytic activity of the intestinal proteasome leading to increased production of antigenic peptides. The high mass accuracy in the low ppm range and resolution of FTICR-MS provide direct identifications of individual proteins as mixtures such as components resulting from incomplete electrophoretic separation. In addition, the comparison of UV- and IR-MALDI FTICR-MS may provide details of fragmentation and rearrangement reactions that may occur under UV-MALDI ionisation conditions. PMID- 19544024 TI - Liquid chromatography-mass spectrometry-based quantitative proteomics. AB - During the last decades, molecular sciences revolutionized biomedical research and gave rise to the biotechnology industry. During the next decades, the application of the quantitative sciences--informatics, physics, chemistry, and engineering--to biomedical research brings about the next revolution that will improve human healthcare and certainly create new technologies, since there is no doubt that small changes can have great effects. It is not a question of "yes" or "no," but of "how much," to make best use of the medical options we will have. In this context, the development of accurate analytical methods must be considered a cornerstone, since the understanding of biological processes will be impossible without information about the minute changes induced in cells by interactions of cell constituents with all sorts of endogenous and exogenous influences and disturbances. The first quantitative techniques, which were developed, allowed monitoring relative changes only, but they clearly showed the significance of the information obtained. The recent advent of techniques claiming to quantify proteins and peptides not only relative to each other, but also in an absolute fashion, promised another quantum leap, since knowing the absolute amount will allow comparing even unrelated species and the definition of parameters will permit to model biological systems much more accurate than before. To bring these promises to life, several approaches are under development at this point in time and this review is focused on those developments. PMID- 19544025 TI - iTRAQ-labeling of in-gel digested proteins for relative quantification. AB - In addition to standard MS-based protein identification, quantification of proteins by mass spectrometry (MS) is rapidly gaining acceptance in proteomic studies. MS-based quantification involves either the incorporation of stable isotopes or can be performed label-free. Recently, more attention has been devoted to label-free quantification; however, this approach has not been fully established among the proteomic community yet. More common is still the introduction of stable isotopes, which can be done by metabolic (e.g., SILAC) or by chemical (e.g., ICAT, iTRAQ, etc.) labeling. Here, we present an overall quantification strategy for chemical labeling of in-gel digested proteins using iTRAQ reagents. This includes (1) protein separation by gel electrophoresis, (2) excision of protein bands, (3) in-gel digestion and extraction of peptides, (4) labeling of peptides, (5) pooling the samples to be compared, (6) LC-MS/MS of labeled peptides, and (7) database search. The presented workflow is well suited for protein samples of moderate complexity (i.e., protein samples of 300-400 components), and it is exemplified by using different amounts of 25S [U4/U6.U5] tri-snRNPs. PMID- 19544027 TI - Algorithms and databases. AB - The capacity of proteomics methods and mass spectrometry instrumentation to generate data has grown substantially over the past years. This data volume growth has in turn led to an increased reliance on software to identify peptide or protein sequences from the recorded mass spectra. Diverse algorithms can be applied for the processing of these data, each performing a specific task such as spectrum quality filtering, spectral clustering and merging, assigning a sequence to a spectrum, and assessing the validity of these assignments. The key algorithms to mass spectral processing pipelines are the ones that assign a sequence to a spectrum. The most commonly used variants of these are crucially dependent on the information contained in the sequences database, which they use as a basis for identification. Since these sequence databases are constructed in different ways and can therefore vary substantially in the amount and type of data they contain, they are also discussed here. PMID- 19544026 TI - Electrospray mass spectrometry for quantitative plasma proteome analysis. AB - Electrospray ionization mass spectrometry (ESI-MS) is an efficient soft ionization procedure for macro biomolecules. However, it is a rather delicate process to produce charged molecules for mass-to-charge ratio (m/z) based measurement. In this chapter, the mechanism of ESI is briefly presented, and the experimental pipeline for quantitative profiling of plasma proteins (prefractionation immunodepletion, protein isotope tagging, 2D-HPLC separation of intact proteins, and LC-MS) is presented as applied by our group in studies of cancer biomarker discovery. PMID- 19544028 TI - Shotgun protein identification and quantification by mass spectrometry. AB - Shotgun proteomics is based on identification and quantification of peptides from digested proteins using tandem mass spectrometry. In this chapter, we discuss computational methods to analyze tandem mass spectra of peptides, including database searching, de novo peptide sequencing, hybrid approaches, library searching, and unrestricted modification search. A special focus is given to database searching programs since they are most widely used. The process of inferring proteins from identified peptides is then discussed. We also provide description of key steps in the quantitative analysis of mass spectrometry proteomics data. PMID- 19544029 TI - Proteomics identification of oxidatively modified proteins in brain. AB - Several studies demonstrated the involvement of free radicals in the pathophysiology of neurodegenerative diseases. Once formed, reactive oxygen species (ROS) can promote multiple forms of oxidative damage, including protein oxidation, and thereby influence the function of a diverse array of cellular processes leading inevitably to neuronal dysfunctions. Protein oxidation can therefore rapidly contribute to oxidative stress by directly affecting cell signaling, cell structure, and enzymatic processes such as metabolism. There are many different modes of inducing protein oxidation including metal-catalyzed oxidation, oxidation-induced cleavage of peptide chain, amino acid oxidation, and the covalent binding of lipid peroxidation products or advanced glycation end proteomics. In this paper we describe the protocol of redox proteomics, a tool to identify post-translational modifications of proteins. We focus our attention on the identification of carbonylated and 4-hydroxy-2-trans-nonenal-bound proteins. In redox proteomics, samples for the identification of protein carbonyls are first derivatized with 2,4-dinitrophenolhydrazine (DNPH) followed by two dimensional (2D) separation of these proteins based on their isoelectric point and rate of migration. The carbonylated proteins are then detected using 2D Western blot techniques. Similarly, HNE-bound proteins can be detected using the above-mentioned strategy except that the sample does not need to be derivatized. Separated proteins are identified following tryptic digestion, mass spectrometry, and interrogation of appropriate databases. PMID- 19544030 TI - Isotope-labeling and affinity enrichment of phosphopeptides for proteomic analysis using liquid chromatography-tandem mass spectrometry. AB - The reversible phosphorylation of proteins is a dynamic process that plays a major role in many vital physiological processes by transmitting signals within cellular pathways and networks. Proteomic measurements using mass spectrometry are capable of characterizing the sites of protein phosphorylation and to quantify their change in abundance. However, the low stoichiometry of protein phosphorylation events often preclude mass spectrometry detection and require additional sample preparation steps to facilitate their characterization. Many analytical methods have been used to map and quantify changes in phosphorylation, and this chapter will present two methods that can be used for extraction of phosphopeptides from protein and proteome digests to map phosphorylation sites using liquid chromatography-tandem mass spectrometry (LC/MS/MS). The first method describes an immobilized metal affinity chromatography (IMAC) technique using Ga3+ to enrich for phosphopeptides from protein digests. The second method describes the utilization of phosphoprotein isotope-coded solid-phase tags (PhIST) to label and enrich phosphopeptides from complex mixtures to both identify and quantify changes in protein phosphorylation. The IMAC and PhIST protocols can be applied to any isolated protein sample and is amenable to additional fractionation using strong cation/anion exchange chromatography prior to reversed-phase LC/MS/MS analysis. PMID- 19544031 TI - Organelle proteomics: reduction of sample complexity by enzymatic in-gel selection of native proteins. AB - One major problem in proteomics is the biochemical complexity of living cells. Therefore, strategies are needed to reduce the number of proteins to a manageable amount, enabling researchers to make a statement concerning protein functions. One possibility is the isolation of organelles, which reduces the protein complexity, e.g., for the chloroplast to an estimated number of 2,700 different proteins. For further limitation of the protein number, proteins can be divided into membrane and soluble proteins, which can be analyzed separately in a subsequent step. For membrane proteins, blue native polyacrylamide gel electrophoresis (BN-PAGE) in combination with enzymatic in-gel assays (e.g. detection of NADPH dehydrogenases) is a suitable method for a fast and easy visualization and identification of only one class of membrane proteins. PMID- 19544032 TI - Isolation of plasma membranes from the nervous system by countercurrent distribution in aqueous polymer two-phase systems. AB - The plasma membrane separates the cell-interior from the cell's environment. To maintain homeostatic conditions and to enable transfer of information, the plasma membrane is equipped with a variety of different proteins such as transporters, channels, and receptors. The kind and number of plasma membrane proteins are a characteristic of each cell type. Owing to their location, plasma membrane proteins also represent a plethora of drug targets. Their importance has entailed many studies aiming at their proteomic identification and characterization. Therefore, protocols are required that enable their purification in high purity and quantity. Here, we report a protocol, based on aqueous polymer two-phase systems, which fulfils these demands. Furthermore, the protocol is time-saving and protects protein structure and function. PMID- 19544033 TI - Enrichment and preparation of plasma membrane proteins from Arabidopsis thaliana for global proteomic analysis using liquid chromatography-tandem mass spectrometry. AB - The plasma membrane proteins are critical components in cellular control and differentiation and thus are of special interest to those studying signal transduction mechanisms in all organisms. When conducting proteomic studies on membrane components of cells and tissues, the complexity is not simply confined to the large number of proteins present in the sample but also to the highly hydrophobic nature of membrane proteins containing multiple transmembrane domains. Consequently, these proteins are more difficult to analyze by mass spectrometry, particularly if protein sequence coverage is to be established. This chapter contains a method for extraction, solubilization, alkylation, proteolysis, and identification of hydrophobic integral plasma membrane proteins for large-scale proteomic analysis using strong cation exchange chromatography (SCXC) and liquid chromatography-tandem mass spectrometry (LC/MS/MS). In our approach, microsomes are isolated from plant tissue and then subjected to a two phase extraction procedure to enrich for plasma membranes. Proteins are extracted and solubilized from the membrane using a methanol-aqueous buffer system that allows for effective reduction, cysteinyl alkylation, and tryptic digestion for subsequent SCXC-LC/MS/MS analysis. Our protocol is also amenable to isotope labeling methods to quantify integral membrane protein expression and post translational modifications. In addition to plants, the method can be applied to other systems quite readily; thus, we anticipate that it will be of general interest to those characterizing plasma membrane proteins of any organism. PMID- 19544034 TI - Tandem affinity purification of protein complexes from mammalian cells by the Strep/FLAG (SF)-TAP tag. AB - Isolation and dissection of native multiprotein complexes is a central theme in functional genomics. The development of the tandem affinity purification (TAP) tag has enabled efficient and large-scale purification of native protein complexes. The SF-TAP tag, a modified version of the TAP tag, allows a fast and straightforward purification of protein complexes from mammalian cells. It consists of a tandem Strep-tag II and a FLAG epitope (SF-TAP). The SF-TAP tag allows a native elution of protein complexes without proteolytic cleavage needed in the original TAP procedure. Besides the SF-TAP protocol, the principal idea of a pathway mapping by subsequent tagging of copurified proteins is demonstrated for the interactome of the MAPKKK Raf. PMID- 19544035 TI - Sequential peptide affinity purification system for the systematic isolation and identification of protein complexes from Escherichia coli. AB - Biochemical purification of affinity-tagged proteins in combination with mass spectrometry methods is increasingly seen as a cornerstone of systems biology, as it allows for the systematic genome-scale characterization of macromolecular protein complexes, representing demarcated sets of stably interacting protein partners. Accurate and sensitive identification of both the specific and shared polypeptide components of distinct complexes requires purification to near homogeneity. To this end, a sequential peptide affinity (SPA) purification system was developed to enable the rapid and efficient isolation of native Escherichia coli protein complexes (J Proteome Res 3:463-468, 2004). SPA purification makes use of a dual-affinity tag, consisting of three modified FLAG sequences (3X FLAG) and a calmodulin binding peptide (CBP), spaced by a cleavage site for tobacco etch virus (TEV) protease (J Proteome Res 3:463-468, 2004). Using the lambda phage Red homologous recombination system (PNAS 97:5978-5983, 2000), a DNA cassette, encoding the SPA-tag and a selectable marker flanked by gene-specific targeting sequences, is introduced into a selected locus in the E. coli chromosome so as to create a C-terminal fusion with the protein of interest. This procedure aims for near-endogenous levels of tagged protein production in the recombinant bacteria to avoid spurious, non-specific protein associations (J Proteome Res 3:463-468, 2004). In this chapter, we describe a detailed, optimized protocol for the tagging, purification, and subsequent mass spectrometry-based identification of the subunits of even low-abundance bacterial protein complexes isolated as part of an ongoing large-scale proteomic study in E. coli (Nature 433:531-537, 2005). PMID- 19544036 TI - Bioinformatical approaches to detect and analyze protein interactions. AB - Protein-protein interactions are the building blocks of cellular networks and at the heart of cellular regulation. However, their experimental identification is still a challenge. This chapter is concerned with the determination of protein protein interactions by bioinformatical methods. These often can operate just on sequence information. Further required information is derived from public knowledge in literature databanks and biochemical databases as well as from the sequences themselves and iterative sequence comparisons. Further tools include domain analysis, structure prediction, and genome context methods. The results are predicted binary interactions and complete interaction networks. PMID- 19544037 TI - Food derived carbonyl compounds affect basal and stimulated secretion of interleukin-6 and -8 in Caco-2 cells. AB - BACKGROUND: The carbonyl compounds methylglyoxal (MG) and glyoxal (G) are reactive intermediates generated in a variety of foods and beverages during processing and prolonged storage. AIM AND METHODS: We investigated direct effects of these compounds on intestinal cells determining the basal and stimulated secretion of IL-8 and IL-6 in vitro. RESULTS: MG or G induced a concentration dependent enhancement of IL-8 and IL-6 secretion compared to baseline levels. A co-incubation with pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) or lipopolysaccharides (LPS) and increasing MG concentrations further enhanced IL-8 and IL-6 secretion. For G, however, this additive effect was only observed in TNF-alpha and IL-1beta treated cells, but not after co-incubation with LPS. CONCLUSION: These results suggest a pro-inflammatory effect of G and MG at high concentrations in human intestinal cells by stimulating IL-8 and IL-6 cytokine levels. Effects of G and MG in combination with other cytokines may negatively affect inflammatory processes. PMID- 19544038 TI - Fine structural quantification of drought-stressed Picea abies (L.) organelles based on 3D reconstructions. AB - Ultrastructural investigations of cells and organelles by transmission electron microscopy (TEM) usually lead to two-dimensional information of cell structures without supplying exact quantitative data due to the limited number of investigated ultrathin sections. This can lead to misinterpretation of observed structures especially in context of their three-dimensional (3D) assembly. 3D investigations and quantitative morphometric analysis are therefore essential to get detailed information about the arrangement and the amount of subcellular structures inside a cell or organelle, respectively, especially when the plant sample was exposed to environmental stress. In the present research, serial sectioned chloroplasts, mitochondria, and peroxisomes from first year spruce needles (Picea abies (L.) Karst.) were 3D reconstructed and digitally measured using a computer-supported image analysis system in order to obtain a detailed quantitative characterization of complete cell organelles including precise morphological data of drought-induced fine structural changes. In control plants, chloroplast volume was composed of 56% stroma, 15% starch, 27% thylakoids, and 2% plastoglobules. In drought-stressed chloroplasts, the relative volume of both the thylakoids and the plastoglobules significantly increased to 37% and 12%, respectively. Chloroplasts of stressed plants differed from control plants not only in the mean thylakoid and plastoglobules content but also in the complete lack of starch grains. Mitochondria occurred in variable forms in both control and stressed samples. In stressed plants, mitochondria showed a significant smaller mean volume which was only 81% when compared with the control organelles. Peroxisomes were inconspicuous in both samples and their volume did not differ between control and drought-stressed samples. The present study shows that specific subcellular structures are subject to significant quantitative changes during drought stress of spruce needles giving a detailed insight in adaptation processes of the investigated cell organelles. PMID- 19544039 TI - Molecular analysis of the gene encoding a cold-adapted halophilic subtilase from deep-sea psychrotolerant bacterium Pseudoalteromonas sp. SM9913: cloning, expression, characterization and function analysis of the C-terminal PPC domains. AB - Only a few cold-adapted halophilic proteases have been reported. Here, the gene mcp03 encoding a cold-adapted halophilic protease MCP-03 was cloned from deep-sea psychrotolerant bacterium Pseudoalteromonas sp. SM9913, which contains a 2,130-bp ORF encoding a novel subtilase precursor. The recombinant MCP-03, expressed in Escherichia coli BL21 and purified from fermented broth, is a multi-domain protein with a catalytic domain and two PPC domains. Compared to mesophilic subtilisin Carlsberg, MCP-03 had characteristics of a typical cold-adapted enzyme (e.g., higher activity at low temperatures, lower optimum temperature and higher thermolability). MCP-03 also exhibited good halophilic ability with maximal activity at 3 M NaCl/KCl and good stability in 3 M NaCl. Deletion mutagenesis showed that the C-terminal PPC domains were unnecessary for enzyme secretion but had an inhibitory effect on MCP-03 catalytic efficiency and were essential for keeping MCP-03 thermostable. PMID- 19544041 TI - Recognition of downhill esophageal varices in hemodialysis patients requires a high index of clinical suspicion. PMID- 19544040 TI - Carboxylic ester hydrolases from hyperthermophiles. AB - Carboxylic ester hydrolyzing enzymes constitute a large group of enzymes that are able to catalyze the hydrolysis, synthesis or transesterification of an ester bond. They can be found in all three domains of life, including the group of hyperthermophilic bacteria and archaea. Esterases from the latter group often exhibit a high intrinsic stability, which makes them of interest them for various biotechnological applications. In this review, we aim to give an overview of all characterized carboxylic ester hydrolases from hyperthermophilic microorganisms and provide details on their substrate specificity, kinetics, optimal catalytic conditions, and stability. Approaches for the discovery of new carboxylic ester hydrolases are described. Special attention is given to the currently characterized hyperthermophilic enzymes with respect to their biochemical properties, 3D structure, and classification. PMID- 19544042 TI - Chiropractic treatment for fibromyalgia: a systematic review. AB - Many patients use chiropractic as a treatment of fibromyalgia, and many chiropractors seem to be convinced that it is effective for that condition. The aim of the study was to conduct a systematic review of randomised clinical trials testing the effectiveness of chiropractic care for fibromyalgia. Six electronic literature searches were conducted. No language restrictions were applied. Data extraction and validation were carried out by two independent reviewers. Three studies met the inclusion criteria. Their methodological quality was poor. They generated no evidence to suggest that chiropractic care is effective for fibromyalgia. Currently, there is insufficient evidence to conclude that chiropractic is an effective treatment for fibromyalgia. PMID- 19544043 TI - Comparison of splinting and splinting plus low-level laser therapy in idiopathic carpal tunnel syndrome. AB - This study aimed to compare the short-term efficacy of splinting (S) and splinting plus low-level laser therapy (SLLLT) in mild or moderate idiopathic carpal tunnel syndrome (CTS) with a prospective, randomized controlled study. The patients with unilateral, mild, or moderate idiopathic CTS who experienced symptoms over 3 months were included in the study. The SLLLT group received ten sessions of laser therapy and splinting while S group was given only splints. The patients were evaluated at the baseline and after 3 months of the treatment. Follow-up parameters were nerve conduction study (NCS), Boston Questionnaire (BQ), grip strength, and clinical response criteria. Forty-five patients with CTS completed the study. Twenty-four patients were in S and 21 patients were in SLLLT group. In the third-month control, SLLLT group had significant improvements on both clinical and NCS parameters (median motor nerve distal latency, median sensory nerve conduction velocities, BQ symptom severity scale, and BQ functional capacity scale) while S group had only symptomatic healing (BQ symptom severity scale). The grip strength of splinting group was decreased significantly. According to clinical response criteria, in SLLLT group, five (23.8%) patients had full and 12 (57.1%) had partial recovery; four (19%) patients had no change or worsened. In S group, one patient (4.2%) had full and 17 (70.8%) partial recovery; six (25%) patients had no change or worsened. Additionally, applied laser therapy provided better outcomes on NCS but not in clinical parameters in patients with CTS. PMID- 19544044 TI - The ATP-binding cassette family: a structural perspective. AB - The ATP-binding cassette family is one of the largest groupings of membrane proteins, moving allocrites across lipid membranes, using energy from ATP. In bacteria, they reside in the inner membrane and are involved in both uptake and export. In eukaryotes, these transporters reside in the cell's internal membranes as well as in the plasma membrane and are unidirectional-out of the cytoplasm. The range of substances that these proteins can transport is huge, which makes them interesting for structure-function studies. Moreover, their abundance in nature has made them targets for structural proteomics consortia. There are eight independent structures for ATP-binding cassette transporters, making this one of the best characterised membrane protein families. Our understanding of the mechanism of transport across membranes and membrane protein structure in general has been enhanced by recent developments for this family. PMID- 19544045 TI - Infection with murine gammaherpesvirus 68 exacerbates inflammatory bowel disease in IL-10-deficient mice. AB - OBJECTIVE: We questioned whether infection with murine gammaherpesvirus 68 (HV 68) might exacerbate inflammatory bowel disease using mice deficient in IL-10 (IL 10-/-) as a model of developing colitis. METHODS: Groups of C57BL/6 mice and IL 10-/- mice were mock-treated or infected with HV-68. Two months following infection, mice were euthanized and a variety of parameters were measured to quantify the extent of inflammation and the presence of virus. Measurements included survival, body weight, splenomegaly, colonic disease scores, liver histopathology, viable bacteria in the liver, and splenic viral burden. RESULTS: IL-10-/- mice infected with HV-68 displayed reduced survival, lower body weights, increased splenomegaly, exacerbated colonic disease scores, increased numbers of viable bacteria in the liver, and increased leukocyte liver infiltration when compared to mock-treated IL-10-/- mice or HV-68 infected C57BL/6 mice. Surprisingly, levels of infectious or latent virus were not significantly different between the groups of mice exposed to HV-68. CONCLUSIONS: The presence of HV-68 in IL-10-/- mice exacerbates the developing clinical disease in this animal model of colitis. PMID- 19544046 TI - Receptor mediation and nociceptin inhibition of bradykinin-induced plasma extravasation in the knee joint of the rat. AB - OBJECTIVE AND DESIGN: The aim was to investigate the signaling mechanisms and regulation of bradykinin (BK)-induced inflammation in rat knee joint. MATERIALS AND METHODS: Knee joints of anesthetized rats were perfused with BK (0.1-1.0 microM), and synovial plasma extravasation (PE) was evaluated by spectrophotometrical measurement of Evans Blue leakage. To examine the signaling pathway, B1 antagonist [des-Arg10]-HOE140 (0.1-1.0 microM) and B2 antagonist HOE140 (0.05-1.0 microM), calcitonin gene-related peptide (CGRP) antagonist CGRP8 37 (0.5-1.0 microM), prostaglandin E2 antagonist AH-6809 (0.1-1.0 microM), and histamine H1 antagonist mepyramine (0.1-1.0 microM) were used. Nociceptin (0.0001 1.0 microM) and antagonist J-113397 were tested for modulation of BK-induced PE. The analyses were compared side-by-side with 5-hydroxytryptamine-induced PE. RESULTS: BK perfusion dose-dependently induced PE, which was blocked by HOE140, CGRP8-37, AH-6809, and mepyramine. It was also inhibited by nociceptin, which could be reversed by antagonist J-113397. In contrast, 5-hydroxytryptamine induced PE was biphasically regulated by nociceptin and was not antagonized by CGRP8-37. CONCLUSIONS: BK-induced PE is mediated by B2 receptors and may involve CGRP, prostaglandin, and histamine pathways. BK-induced PE is inhibited by nociceptin through the activation of ORL1 receptors. There are differences between BK- and 5-hydroxytryptamine-induced inflammation in signaling and modulation. PMID- 19544047 TI - Overexpression of two chrysanthemum DgDREB1 group genes causing delayed flowering or dwarfism in Arabidopsis. AB - We isolated 13 DREB1 (dehydration responsive element binding factor 1) genes from chrysanthemum and further divided them into three groups, DgDREB1A, DgDREB1B and DgDREB1C, based on the phylogenetic analysis. Each group showed their unique expression patterns under cold, dehydration and salt stress conditions. Arabidopsis plants overexpressing DgDREB1A (1A plants) exhibited significantly stronger tolerance to freezing and drought than those overexpressing DgDREB1B (1B plants) and the control plants. In addition, 1A plants showed delayed flowering, but not dwarfism; while 1B plants showed dwarfism, but not delayed flowering. In 1A plants, the expression of three stress-related DREB1-downstream genes, COR47, COR15A, and RD29A, was strongly induced while the expression of CO and FT, two photoperiod responsive flowering-time genes, was inhibited. In 1B plants, the expression of GA2ox7, a GA-deactivation enzyme gene, was dramatically enhanced. The results above strongly suggest that members from different DgDREB1 groups may have distinct effects on plant development: DgDREB1A may be involved in photoperiod-related flowering-time determination and DgDREB1B in GA-mediated plant development. PMID- 19544051 TI - Integrated genetic map and genetic analysis of a region associated with root traits on the short arm of rye chromosome 1 in bread wheat. AB - A rye-wheat centric chromosome translocation 1RS.1BL has been widely used in wheat breeding programs around the world. Increased yield of translocation lines was probably a consequence of increased root biomass. In an effort to map loci controlling root characteristics, homoeologous recombinants of 1RS with 1BS were used to generate a consensus genetic map comprised of 20 phenotypic and molecular markers, with an average spacing of 2.5 cM. Physically, all recombination events were located in the distal 40% of the arms. A total of 68 recombinants was used and recombination breakpoints were aligned and ordered over map intervals with all the markers, integrated together in a genetic map. This approach enabled dissection of genetic components of quantitative traits, such as root traits, present on 1S. To validate our hypothesis, phenotyping of 45-day-old wheat roots was performed in five lines including three recombinants representative of the entire short arm along with bread wheat parents 'Pavon 76' and Pavon 1RS.1BL. Individual root characteristics were ranked and the genotypic rank sums were subjected to Quade analysis to compare the overall rooting ability of the genotypes. It appears that the terminal 15% of the rye 1RS arm carries gene(s) for greater rooting ability in wheat. PMID- 19544048 TI - Effect of CaCl2 hydrothermal treatment on the bone bond strength and osteoconductivity of Ti-0.5Pt and Ti-6Al-4V-0.5Pt alloy implants. AB - To achieve osteoconductivity, Ti-0.5Pt and Ti-6Al-4V-0.5Pt alloys were hydrothermally treated at 200 degrees C in 10 mmol/l CaCl(2) aqueous solution for 24 h (HT-treatment). We conducted histological investigations of the HT-treated materials by using Wistar strain rats (SD rats) to evaluate the usefulness of the treatment. To measure the bone bond strength, the specimens were implanted in the tibia of SD rats, and a pull-out test was conducted. From the early postoperative stages, direct bone contact was obtained for the HT-treated implants. Within 1-4 weeks of implantation, the bone contact ratios and bone bond strengths of the HT treated implants were higher than those of the non-treated implants. The Ti-0.5Pt and Ti-6Al-4V-0.5Pt alloys with HT-treatment showed the potential to develop a new implant with a high bone bond strength and rapid osteoconduction. PMID- 19544053 TI - Visual cueing with context relevant information for reducing change blindness. AB - OBJECTIVE: Physiological monitoring is a requisite for optimal care to ensure that the condition of a patient is maintained within safe levels. Monitoring can be jeopardized by the inability of a clinician to recognize important changes in the visual display of data throughout the duration of the monitoring task. We hypothesized that the addition of a visual cue imparting contextual information to a physiological display would improve the detection ability and response time of a clinician to a change in a patient variable. METHODS: Contextual information based on trend information was added to a physiological display in the form of a visual cue. Following IRB approval, the resulting enhanced display was evaluated by 22 anesthesiologists in a simulated operating room, through the observation of six simulated scenarios using a standard anesthesia display and the enhanced display. Demographic information, response time, accuracy of detection, and usability data were collected. RESULTS: The enhanced display reduced the detection time to a change in the simulated scenarios by 14.4 s (95% CI: -26.4 to -2.38), and reduced the expected number of missed events per scenario by 0.23 (95% CI: -0.439 to -0.0203), based on the repeated measures analysis (Poisson model). CONCLUSIONS: The data collected and analyzed in this study supports the addition of a visual cue to future physiological monitors. The graphic representation and the context relevant information that it transmits appears to aid clinicians. While the results indicate that enhanced visualization of context relevant information can lead to a significant improvement in event recognition and identification, further evaluation in clinical settings is required. PMID- 19544055 TI - Effect of Fusarium oxysporum f. sp. lycopersici on the degradation of humic acid associated with Cu, Pb, and Ni: an in vitro study. AB - The intent of this work was to gain further insight on the fungus-assisted degradation/solubilization of humic acid and the related changes in metal-binding profiles. In the experimental design, Aldrich reagent humic acid (HA) or HA enriched with Cu, Pb, and Ni (HA(Me)) was added to Fusarium oxysporum f. sp. lycopersici cultures in vitro. The cultures were supplied by different carbon- and nitrogen-containing nutrients (glucose, Glc, or glutamate, Glu and ammonium, NH4+, or nitrate, NO3-, ions, respectively) in order to examine their possible effect on HA and HA(Me) decomposition. During the first 48 h of fungus growth, gradual acidification to pH 2 was observed in medium containing Glc + NH4+, while for other cultures, alkalinization to pH 9 occurred and then, the above conditions were stable up to at least 200 h. Size exclusion chromatography (SEC) with UV/Vis detection showed progressive degradation and solubilization of both HA and HA(Me) with the increasing time of fungus growth. However, the molecular mass distributions of HA-related soluble species were different in the presence of metals (HA(Me)) as referred to HA and were also influenced by the composition of growth medium. The solubilization of Pb, Cu, and Ni and their association with HA molecular mass fractions were studied using inductively coupled plasma mass spectrometry (ICP-MS) detection. Under acidic conditions, relatively high concentrations of low-molecular-mass metallic species were found in culture supernatants, while in alkaline media, metal solubilization was generally poorer. In contrast to low pH culture, SEC-ICP-MS results obtained in alkaline supernatants indicated metal binding to degradation products of humic substances of MM > 5 kDa. In summary, the results of this study suggest that fungus-assisted degradation of HA and HA(Me) might be controlled using appropriate N- and C- sources required for fungus growth, which in turn would affect molecular mass distribution of soluble metallic species thus potentially influencing their actual bioaccessibility. PMID- 19544054 TI - Quantitative analysis of methyl green using surface-enhanced resonance Raman scattering. AB - Surface-enhanced resonance Raman scattering (SERRS) spectra of aqueous solutions of the triphenylmethane dye methyl green have been obtained for the first time by use of citrate-reduced silver colloids and a laser excitation wavelength of 632.8 nm. Given the highly fluorescent nature of the analyte, which precluded collection of normal Raman spectra of the dye in solution and powdered state, it was highly encouraging that SERRS spectra showed no fluorescence due to quenching by the silver sol. The pH conditions for SERRS were optimised over the pH range 0.5-10 and the biggest enhancement for SERRS of this charged dye was found to be at pH 2.02, thus this condition was used for quantitative analysis. SERRS was found to be highly sensitive and enabled quantitative determination of methyl green over the range 10(-9) to 10(-7) mol dm(-3). Good fits to correlation coefficients were obtained over this range using the areas under the vibrational bands at 1615 and 737 cm(-1). Finally, a limit of detection of 83 ppb was calculated, demonstrating the sensitivity of the technique. PMID- 19544052 TI - Subacute exposure to N-ethyl perfluorooctanesulfonamidoethanol results in the formation of perfluorooctanesulfonate and alters superoxide dismutase activity in female rats. AB - Perfluorooctanesulfonamides, such as N-ethyl perfluorooctanesulfonamidoethanol (N EtFOSE), are large scale industrial chemicals but their disposition and toxicity are poorly understood despite significant human exposure. The hypothesis that subacute exposure to N-EtFOSE, a weak peroxisome proliferator, causes a redox imbalance in vivo was tested using the known peroxisome proliferator, ciprofibrate, as a positive control. Female Sprague-Dawley rats were treated orally with N-EtFOSE, ciprofibrate or corn oil (vehicle) for 21 days, and levels of N-EtFOSE and its metabolites as well as markers of peroxisome proliferation and oxidative stress were assessed in serum, liver and/or uterus. The N-EtFOSE metabolite profile in liver and serum was in good agreement with reported in vitro biotransformation pathways in rats and the metabolite levels decreasing in the order perfluorooctanesulfonate >> perfluorooctanesulfonamide ~ N-ethyl perfluorooctanesulfonamidoacetate >> perfluorooctanesulfonamidoethanol approximately N-EtFOSE. Although N-EtFOSE treatment significantly decreased the growth rate, increased relative liver weight and activity of superoxide dismutases (SOD) in liver and uterus (total SOD, CuZnSOD and MnSOD), a metabolic study revealed no differences in the metabolome in serum from N-EtFOSE-treated and control animals. Ciprofibrate treatment increased liver weight and peroxisomal acyl Co-A oxidase activity in the liver and altered antioxidant enzyme activities in the uterus and liver. According to NMR metabolomic studies, ciprofibrate treated animals had altered serum lipid profiles compared to N EtFOSE-treated and control animals, whereas putative markers of peroxisome proliferation in serum were not affected. Overall, this study demonstrates the biotransformation of N-EtFOSE to PFOS in rats that is accompanied by N-EtFOSE induced alterations in antioxidant enzyme activity. PMID- 19544056 TI - Determination of the polar pesticide degradation product N,N-dimethylsulfamide in aqueous matrices by UPLC-MS/MS. AB - This study presents a fast, sensitive, and robust method for the determination of the polar pesticide degradation product N,N-dimethylsulfamide (DMS) in water based on ultraperformance liquid chromatography-tandem mass spectrometry (UPLC MS/MS). To provide a robust analysis method, the use of an internal standard for both natural waters and model water was examined in order to compensate for matrix effects. The relative standard deviation was found to be +/-15% (n = 10) and the limit of detection was 10 ng/L by direct injection in the UPLC-MS/MS system. The only sample preparation step required is the addition of the internal standard. The chromatographic analysis of one sample takes 4 min and thus is applicable for economic routine laboratory work. More than 600 samples of drinking water, surface water, and groundwater have been examined successfully with this method in the Rhine and Ruhr region of North Rhine Westphalia (Germany). Approximately 65% of analyzed samples contained measurable amounts of DMS at concentrations up to 63 microg/L. PMID- 19544057 TI - Pointing control using a moving base of support. AB - The purposes of this study were to determine whether gaze direction provides a control signal for movement direction for a pointing task requiring a step and to gain insight into why discrepancies previously identified in the literature for endpoint accuracy with gaze directed eccentrically exist. Straight arm pointing movements were performed to real and remembered target locations, either toward or 30 degrees eccentric to gaze direction. Pointing occurred in normal room lighting or darkness while subjects sat, stood still or side-stepped left or right. Trunk rotation contributed 22-65% to gaze orientations when it was not constrained. Error differences for different target locations explained discrepancies among previous experiments. Variable pointing errors were influenced by gaze direction, while mean systematic pointing errors and trunk orientations were influenced by step direction. These data support the use of a control strategy that relies on gaze direction and equilibrium inputs for whole body goal-directed movements. PMID- 19544058 TI - A multisensory approach to spatial updating: the case of mental rotations. AB - Mental rotation is the capacity to predict the outcome of spatial relationships after a change in viewpoint. These changes arise either from the rotation of the test object array or from the rotation of the observer. Previous studies showed that the cognitive cost of mental rotations is reduced when viewpoint changes result from the observer's motion, which was explained by the spatial updating mechanism involved during self-motion. However, little is known about how various sensory cues available might contribute to the updating performance. We used a Virtual Reality setup in a series of experiments to investigate table-top mental rotations under different combinations of modalities among vision, body and audition. We found that mental rotation performance gradually improved when adding sensory cues to the moving observer (from None to Body or Vision and then to Body & Audition or Body & Vision), but that the processing time drops to the same level for any of the sensory contexts. These results are discussed in terms of an additive contribution when sensory modalities are co-activated to the spatial updating mechanism involved during self-motion. Interestingly, this multisensory approach can account for different findings reported in the literature. PMID- 19544059 TI - Removal of visual feedback alters muscle activity and reduces force variability during constant isometric contractions. AB - The purpose of this study was to compare force accuracy, force variability and muscle activity during constant isometric contractions at different force levels with and without visual feedback and at different feedback gains. In experiment 1, subjects were instructed to accurately match the target force at 2, 15, 30, 50, and 70% of their maximal isometric force with abduction of the index finger and maintain their force even in the absence of visual feedback. Each trial lasted 22 s and visual feedback was removed from 8-12 to 16-20 s. Each subject performed 6 trials at each target force, half with visual gain of 51.2 pixels/N and the rest with a visual gain of 12.8 pixels/N. Force error was calculated as the root mean square error of the force trace from the target line. Force variability was quantified as the standard deviation and coefficient of variation (CVF) of the force trace. The EMG activity of the agonist (first dorsal interosseus; FDI) was measured with bipolar surface electrodes placed distal to the innervation zone. Independent of visual gain and force level, subjects exhibited lower force error with the visual feedback condition (2.53 +/- 2.95 vs. 2.71 +/- 2.97 N; P < 0.01); whereas, force variability was lower when visual feedback was removed (CVF: 4.06 +/- 3.11 vs. 4.47 +/- 3.14, P < 0.01). The EMG activity of the FDI muscle was higher during the visual feedback condition and this difference increased especially at higher force levels (70%: 370 +/- 149 vs. 350 +/- 143 microV, P < 0.01). Experiment 2 examined whether the findings of experiment 1 were driven by the higher force levels and proximity in the gain of visual feedback. Subjects performed constant isometric contractions with the abduction of the index finger at an absolute force of 2 N, with two distinct feedback gains of 15 and 3,000 pixels/N. In agreement with the findings of experiment 1, subjects exhibited lower force error in the presence of visual feedback especially when the feedback gain was high (0.057 +/- 0.03 vs. 0.095 +/- 0.05 N). However, force variability was not affected by the vastly distinct feedback gains at this force, which supported and extended the findings from experiment 1. Our findings demonstrate that although removal of visual feedback amplifies force error, it can reduce force variability during constant isometric contractions due to an altered activation of the primary agonist muscle most likely at moderate force levels in young adults. PMID- 19544060 TI - Grasping isoluminant stimuli. AB - We used a virtual reality setup to let participants grasp discs, which differed in luminance, chromaticity and size. Current theories on perception and action propose a division of labor in the brain into a color proficient perception pathway and a less color-capable action pathway. In this study, we addressed the question whether isoluminant stimuli, which provide only a chromatic but no luminance contrast for action planning, are harder to grasp than stimuli providing luminance contrast or both kinds of contrast. Although we found that grasps of isoluminant stimuli had a slightly steeper slope relating the maximum grip aperture to disc size, all other measures of grip quality were unaffected. Overall, our results do not support the view that isoluminance of stimulus and background impedes the planning of a grasping movement. PMID- 19544061 TI - Effect of glucosamine or chondroitin sulfate on the osteoarthritis progression: a meta-analysis. AB - The aim of this study was to assess the structural efficacies of daily glucosamine sulfate and chondroitin sulfate in patients with knee osteoarthritis (OA). The authors surveyed randomized controlled studies that examined the effects of long-term daily glucosamine sulfate and chondroitin sulfate on joint space narrowing (JSN) in knee OA patients using the Medline and the Cochrane Controlled Trials Register, and by performing manual searches. Meta-analysis was performed using a fixed effect model because no between-study heterogeneity was evident. Six studies involving 1,502 cases were included in this meta-analysis, which consisted of two studies on glucosamine sulfate and four studies on chondroitin sulfate. Glucosamine sulfate did not show a significant effect versus controls on minimum JSN over the first year of treatment (SMD 0.078, 95% CI 0.116 to -0.273, P = 0.429). However, after 3 years of treatment, glucosamine sulfate revealed a small to moderate protective effect on minimum JSN (SMD 0.432, 95% CI 0.235-0.628, P < 0.001). The same was observed for chondroitin sulfate, which had a small but significant protective effect on minimum JSN after 2 years (SMD 0.261, 95% CI 0.131-0.392, P < 0.001). This meta-analysis of available data shows that glucosamine and chondroitin sulfate may delay radiological progression of OA of the knee after daily administration for over 2 or 3 years. PMID- 19544062 TI - Temperature acclimation has no effect on ryanodine receptor expression or subcellular localization in rainbow trout heart. AB - In cardiomyocytes, ryanodine receptors (RYRs) mediate Ca(2+)-induced Ca(2+) release (CICR) from the sarcoplasmic reticulum (SR) during excitation-contraction (e-c) coupling. In rainbow trout heart, the relative importance of CICR increases with cold-acclimation. Thus, the aim of this study was to investigate the effect of temperature acclimation (4, 11 and 18 degrees C) on RYR intracellular localization and expression density. We used immunocytochemistry to assess intracellular localization in ventricular myocytes and Western blotting to assess RYR expression in both atrial and ventricular tissue. In ventricular myocytes, RYRs were localized peripherally in transverse bands aligning with sarcomeric m lines and centrally around mitochondria and the nucleus. Localization did not change with temperature acclimation. RYR expression was also unaffected by temperature acclimation. The localization of RYRs at the m-line is similar to neonatal mammalian cardiomyocytes. We suggest this positioning is indicative of myocytes which rely predominantly on transsarcolemmal Ca(2+)-influx, rather than CICR, during e-c coupling. PMID- 19544063 TI - Changes after voice therapy in objective and subjective voice measurements of pediatric patients with vocal nodules. AB - The aim of this study was to analyze the efficiency of the voice therapy in children with vocal nodules by using the acoustic analysis and subjective assessment. Thirty-nine patients with vocal fold nodules, aged between 7 and 14, were included in the study. Each subject had voice therapy led by an experienced voice therapist once a week. All diagnostic and follow-up workouts were performed before the voice therapy and after the third or the sixth month. Transoral and/or transnasal videostroboscopic examination and acoustic analysis were achieved using multi-dimensional voice program (MDVP) and subjective analysis with GRBAS scale. As for the perceptual assessment, the difference was significant for four parameters out of five. A significant improvement was found in the acoustic analysis parameters of jitter, shimmer, and noise-to-harmonic ratio. The voice therapy which was planned according to patients' needs, age, compliance and response to therapy had positive effects on pediatric patients with vocal nodules. Acoustic analysis and GRBAS may be used successfully in the follow-up of pediatric vocal nodule treatment. PMID- 19544064 TI - Image analyses of conjunctival hyperemia in guinea pig allergic conjunctivitis. PMID- 19544065 TI - Equivalent tamponade by room air as compared with SF(6) after macular hole surgery. AB - BACKGROUND: To evaluate the effect of tamponade by room air after vitrectomy for the treatment of idiopathic macular hole (MH). METHODS: There were 156 eyes of 151 patients studied. The patients' ages ranged from 35 to 88 years old (mean: 65.1 years). After conventional pars plana vitrectomy with internal limiting membrane peeling, fluid air exchange was performed using 20% SF(6) (Gas group: 91 eyes) or room air (Air group: 65 eyes). Surgical outcomes were retrospectively analyzed. RESULTS: Mean preoperative hole diameter was 352 microm in the Gas group and 370 microm in the Air group (P = 0.558). The closure rate of all cases was 91.0% after first surgery and 98.7% at last follow-up. The primary closure rate was 90.1% in the Gas group after 7.44 +/- 1.66 (mean +/- SD) days prone positioning period, and 92.3% in the Air group after 3.83 +/- 0.97 days of prone positioning. There was significant difference in prone positioning period (P < 0.0001), but not in the first closure rate (P = 0.132). CONCLUSION: This study suggests that room air may have an equivalent tamponade effect, in spite of the shorter prone positioning period, than SF(6) after MH surgery. PMID- 19544066 TI - The murine AE4 promoter predominantly drives type B intercalated cell specific transcription. AB - AE4 is an anion exchanger almost exclusively expressed in the collecting ducts of the kidney. This very restricted expression prompted us to analyze its transcription in more detail. 5' RACE yielded alternative transcriptional start sites that are predicted to code for N-terminal protein variants. Comparison of the 5' genomic sequence between species identified a transcriptionally active region with three conserved spans. In transgenic mice beta-galactosidase expression driven by this fragment resembled endogenous AE4 expression and was predominantly restricted to type B intercalated cells. Hence this promoter could prove useful to target type B intercalated cells by genetic approaches. PMID- 19544067 TI - Relationships between surface EMG variables and motor unit firing rates. AB - Although surface electromyography (sEMG) is a widely used electrophysiological technique, its physiological interpretation remains somewhat controversial. This study examined the relationship between motor unit firing rates (MUFR) and the root mean square (RMS) amplitude and mean power frequency (MPF) of the sEMG signal in the biceps brachii. Eleven subjects performed maximal isometric elbow flexion while indwelling and sEMG recordings were obtained from the biceps. The RMS amplitude and MPF of the surface signal, and the mean MUFR from the indwelling signal, were calculated over 500 ms epochs. Group means showed a strong MUFR-RMS amplitude relationship (r (2) = 0.91), but a weak MUFR-MPF relationship (r (2) = 0.20). Using all trials, the MUFR-RMS amplitude (r (2) = 0.19) and MUFR-MPF (r (2) = 0.0037) relationships were much weaker. Within individual subjects, the MUFR-RMS amplitude (mean r (2) = 0.13 +/- 0.17) and the MUFR-MPF (mean r (2) = 0.040 +/- 0.041) relationships were also weak. These results suggest that MUFR cannot be predicted from the characteristics of the sEMG signal. PMID- 19544068 TI - Sensitivity of HCN channel deactivation to cAMP is amplified by an S4 mutation combined with activation mode shift. AB - Hyperpolarisation-activation of HCN ion channels relies on the movement of a charged S4 transmembrane helix, preferentially stabilising the open conformation of the ion pore gate. The open state is additionally stabilised, (a) when cyclic AMP (cAMP) is bound to a cytoplasmic C-terminal domain or (b) when the "mode I" open state formed initially by gate opening undergoes a "mode shift" into a "mode II" open state with a new S4 conformation. We isolated a mutation (lysine 381 to glutamate) in S4 of mouse HCN4; patch-clamp of homomeric channels in excised inside-out membranes revealed a conditional phenotype. When cAMP-liganded K381E channels are previously activated by hyperpolarisation, tens of seconds are required for complete deactivation at a weakly depolarised potential; this "ultra sustained activation" is not observed without cAMP. Whilst cAMP slows deactivation of wild-type channels, the K381E mutation amplifies this effect to enable extraordinary kinetic stabilisation of the open state. K381E channels retain S4-gate coupling, with strong voltage dependence of the rate-limiting step for deactivation of mode II channels near -40 mV. At these voltages, the mode I deactivation pathway shows a different rate-limiting step, lacking strong voltage or cAMP dependence. Ultra-sustained activation thus reflects stabilisation of the mode II open state by the K381E mutation in synergistic combination with cAMP binding. Thus, the voltage-sensing domain is subject to strong functional coupling not only to the pore domain but also to the cytoplasmic cAMP-sensing domain in a manner specific to the voltage sensor conformation. PMID- 19544069 TI - On the mechanism of apoplastic H2O2 production during lignin formation and elicitation in cultured spruce cells--peroxidases after elicitation. AB - A cell culture of Picea abies (L.) Karst. was used for studies of H(2)O(2) generation during constitutive extracellular lignin formation and after elicitation by cell wall fragments of a pathogenic fungus, Heterobasidium parviporum. Stable, micromolar levels of H(2)O(2) were present in the culture medium during lignin formation. Elicitation induced a burst of H(2)O(2), peaking at ca. 90 min after elicitation. Of exogenous reducing substrates that may be responsible for the synthesis of H(2)O(2) from O(2), NADH stimulated H(2)O(2) production irrespective of elicitation. Cysteine (Cys) and glutathione (GSH) partially scavenged the constitutive H(2)O(2), but usually increased or prolonged elicitor-induced H(2)O(2) formation. Culture medium peroxidases were not able to generate H(2)O(2) in vitro with Cys or GSH as reductants. These thiols, however, generated H(2)O(2) non-enzymically at pH 4.5. [(35)S]Sulphate feeding to spruce cells showed that endogenous sulphur-containing compounds (including GSH, GSSG and cysteic acid) existed in the culture medium. The apoplastic levels of these were, however, undetectable by the monobromobimane method suggesting that their contribution to apoplastic H(2)O(2) formation is probably minor. Azide, an inhibitor of haem-containing enzymes, slightly inhibited constitutive H(2)O(2) generation but strongly delayed the elicitor-induced H(2)O(2) accumulation. Diphenylene iodonium, an inhibitor of flavin-containing enzymes, efficiently inhibited H(2)O(2) production irrespective of elicitation. Elicitation led to downregulation of the expression of several peroxidase genes, and peroxidase activity in the culture medium was slightly reduced. Expression of three other peroxidase genes and a respiratory burst oxidase homologue (rboh) gene were upregulated. These data suggest that both peroxidases and rboh may contribute to H(2)O(2) generation. PMID- 19544070 TI - Distribution and phylogeny of the blue light receptors aureochromes in eukaryotes. AB - The new type blue light (BL) receptor aureochrome (AUREO) was recently discovered in a stramenopile alga, Vaucheria (Takahashi et al. Proc Natl Acad Sci USA 104(49):19625-19630, 2007). AUREO has a bZIP (basic region/leucine zipper) and BL sensing light-oxygen-voltage (LOV) domain and functions as a BL-activated transcription factor. It mediates BL-induced branching and regulates the development of the sex organ in V. frigida. Although AUREO sequences have previously been found in Fucus and some diatoms, here we report that AUREO orthologs are commonly conserved in photosynthetic stramenopiles. Five AUREO orthologs were isolated from three stramenopile genera (Fucus, Ochromonas, and Chattonella). By BLAST search, several AUREO sequences were also detected in genomes in Aureococcus anophagefferens (Pelagophyceae). However, AUREO was not found in heterotrophic stramenopiles or in closely related phyla, such as haptophytes and cryptophytes, or in green plants. Stramenopiles do not possess phototropin, the well-known BL receptor for phototropism of green plants. From comparative analysis of LOV domains, together with kinship analysis of AUREO bZIP domains, AUREO can be regarded as the BL receptor specific to phototrophic stramenopiles. The evolution of AUREO and the phylogeny of LOV domains in stramenopiles and green plants are discussed. PMID- 19544071 TI - Surveillance study of apparent life-threatening events (ALTE) in the Netherlands. AB - SIDS and ALTE are different entities that somehow show some similarities. Both constitute heterogeneous conditions. The Netherlands is a low-incidence country for SIDS. To study whether the same would hold for ALTE, we studied the incidence, etiology, and current treatment of ALTE in The Netherlands. Using the Dutch Pediatric Surveillance Unit, pediatricians working in second- and third level hospitals in the Netherlands were asked to report any case of ALTE presented in their hospital from January 2002 to January 2003. A questionnaire was subsequently sent to collect personal data, data on pregnancy and birth, condition preceding the incident, the incident itself, condition after the incident, investigations performed, monitoring or treatment initiated during admission, any diagnosis made at discharge, and treatment or parental support offered after discharge. A total of 115 cases of ALTE were reported, of which 110 questionnaires were filled in and returned (response rate 97%). Based on the national birth rate of 200,000, the incidence of ALTE amounted 0.58/1,000 live born infants. No deaths occurred. Clinical diagnoses could be assessed in 58.2%. Most frequent diagnoses were (percentages of the total of 110 cases) gastro esophageal reflux and respiratory tract infection (37.3% and 8.2%, respectively); main symptoms were change of color and muscle tone, choking, and gagging. The differences in diagnoses are heterogeneous. In 34%, parents shook their infants, which is alarmingly high. Pre- and postmature infants were overrepresented in this survey (29.5% and 8.2%, respectively). Ten percent had recurrent ALTE. In total, 15.5% of the infants were discharged with a home monitor. In conclusion, ALTE has a low incidence in second- and third-level hospitals in the Netherlands. Parents should be systematically informed about the possible devastating effects of shaking an infant. Careful history taking and targeted additional investigations are of utmost importance. PMID- 19544072 TI - Isolation and characterization of microsatellite markers from the Mediterranean fruit fly, Ceratitis capitata: cross-species amplification in other Tephritidae species reveals a varying degree of transferability. AB - The Mediterranean fruit fly, Ceratitis capitata, is a pest of major economic importance and has become a model for the development of SIT control programs for insect pests. Significant information has been accumulated on classical and population genetics of this species during the past 2 decades. However, the availability of molecular markers is limited. Here, we present the isolation and characterization of 159 microsatellite clones and the development of 108 polymorphic microsatellite markers for this insect pest. Mapping by in situ hybridization to polytene chromosomes of 21 microsatellite clones enriched the cytogenetic map that was previously constructed by our group. The enriched map provides a large number of STSs for future genome mapping projects. Cross-species amplification of these microsatellite loci in 12 Tephritidae species and sequence analysis of several amplification products indicated a varying degree of transferability and their possible usefulness as molecular and genetic markers in these species where genetic and molecular tools are limited. PMID- 19544074 TI - Acorn crop size and pre-dispersal predation determine inter-specific differences in the recruitment of co-occurring oaks. AB - The contribution of pre-dispersal seed predation to inter-specific differences in recruitment remains elusive. In species with no resistance mechanisms, differences in pre-dispersal predation may arise from differences in seed abundance (plant satiation) or in the ability of seeds to survive insect infestation (seed satiation). This study aimed to analyse the impact of pre dispersal acorn predation by weevils in two co-occurring Mediterranean oaks (Quercus ilex and Quercus humilis) and to compare its relevance with other processes involved in recruitment. We monitored the patterns of acorn production and acorn infestation by weevils and we conducted experimental tests of acorn germination after weevil infestation, post-dispersal predation and seedling establishment in mixed forests. Monitoring and experimental data were integrated in a simulation model to test for the effects of pre-dispersal predation in recruitment. In both oaks pre-dispersal acorn infestation decreased with increasing acorn crop size (plant satiation). This benefited Q. ilex which exhibited stronger masting behaviour than Q. humilis, with almost a single and outstanding reproductive event in 6 years. Acorn infestation was more than twice as high in Q. humilis (47.0%) as in Q. ilex (20.0%) irrespective of the number of seeds produced by each species. Although germination of infested acorns (seed satiation) was higher in Q. humilis (60%) than in Q. ilex (21%), this could barely mitigate the higher infestation rate in the former species, to reduce seed loss. Conversely to pre-dispersal predation, no inter-specific differences were observed either in post-dispersal predation or seedling establishment. Our results indicate that pre-dispersal predation may contribute to differences in seed supply, and ultimately in recruitment, between co-existing oaks. Moreover, they suggest that seed satiation can barely offset differences in seed infestation rates. This serves as a warning against overemphasising seed satiation as a mechanism to overcome seed predation by insects. PMID- 19544073 TI - Relationship between reversed sexual dimorphism, breeding investment and foraging ecology in a pelagic seabird, the masked booby. AB - Reversed sexual dimorphism (RSD) may be related to different roles in breeding investment and/or foraging, but little information is available on foraging ecology. We studied the foraging behaviour and parental investment by male and female masked boobies, a species with RSD, by combining studies of foraging ecology using miniaturised activity and GPS data loggers of nest attendance, with an experimental study where flight costs were increased. Males attended the chick more often than females, but females provided more food to the chick than males. Males and females foraged during similar periods of the day, had similar prey types and sizes, diving depths, durations of foraging trips, foraging zones and ranges. Females spent a smaller proportion of the foraging trip sitting on the water and had higher diving rate than males, suggesting higher foraging effort by females. In females, trip duration correlated with mass at departure, suggesting a flexible investment through control by body mass. The experimental study showed that handicapped females and female partners of handicapped males lost mass compared to control birds, whereas there was no difference for males. These results indicate that the larger female is the main provisioner of the chick in the pair, and regulates breeding effort in relation to its own body mass, whereas males have a fixed investment. The different breeding investment between the sexes is associated with contrasting foraging strategies, but no clear niche differentiation was observed. The larger size of the females may be advantageous for provisioning the chick with large quantities of energy and for flexible breeding effort, while the smaller male invests in territory defence and nest guarding, a crucial task when breeding at high densities. In masked boobies, division of labour appears to be maximal during chick rearin-g-the most energy demanding period--and may be related to evolution of RSD. PMID- 19544076 TI - Variation of reaction dynamics for OH hydrogen abstraction from glycine between ab initio levels of theory. AB - The variation in reaction dynamics of OH hydrogen abstraction from glycine between HF, MP2, CCSD(T), M05-2X, BHandHLYP, and B3LYP levels was demonstrated. The abstraction mode shows distinct patterns between these five levels and determines the barrier height, and the spin density transfer between OH radical and glycine. These differences are mainly resulted from the spin density distribution and geometry of the alpha carbon during the abstraction. The captodative effect which is commonly believed as one of the major factors to stabilize the caron-centered radical can only be observed in DFT levels but not in HF and MP2 levels. Difference in the abstraction energy were found in these calculation levels, by using the result of CCSD(T) as reference, B3LYP, BHandHLYP, and M05-2X underestimated the reaction barrier about 5.1, 0.1, and 2.4 kcal mol(-1), while HF and MP2 overestimated 19.1 kcal mol(-1) and 1.6 kcal mol( 1), respectively. These differences can be characterized by the vibration mode of imaginary frequency of transition states, which indicates the topology around transition states and determines reaction barrier height. In this model system, BHandHLYP provides the best prediction of the energy barrier among those tested methods. PMID- 19544075 TI - What is an acceptable outcome of treatment before it begins? Methodological considerations and implications for patients with chronic low back pain. AB - Understanding changes in patient-reported outcomes is indispensable for interpretation of results from clinical studies. As a consequence the term "minimal clinically important difference" (MCID) was coined in the late 1980s to ease classification of patients into improved, not changed or deteriorated. Several methodological categories have been developed determining the MCID, however, all are subject to weaknesses or biases reducing the validity of the reported MCID. The objective of this study was to determine the reproducibility and validity of a novel method for estimating low back pain (LBP) patients' view of an acceptable change (MCID(pre)) before treatment begins. One-hundred and forty-seven patients with chronic LBP were recruited from an out-patient hospital back pain unit and followed over an 8-week period. Original and modified versions of the Oswestry disability index (ODI), Bournemouth questionnaire (BQ) and numeric pain rating scale (NRS(pain)) were filled in at baseline. The modified questionnaires determined what the patient considered an acceptable post treatment outcome which allowed us to calculate the MCID(pre). Concurrent comparisons between the MCID(pre), instrument measurement error and a retrospective approach of establishing the minimal clinically important difference (MCID(post)) were made. The results showed the prospective acceptable outcome method scores to have acceptable reproducibility outside measurement error. MCID(pre) was 4.5 larger for the ODI and 1.5 times larger for BQ and NRS(pain) compared to the MCID(post). Furthermore, MCID(pre) and patients post treatment acceptable change was almost equal for the NRS(pain) but not for the ODI and BQ. In conclusion, chronic LBP patients have a reasonably realistic idea of an acceptable change in pain, but probably an overly optimistic view of changes in functional and psychological/affective domains before treatment begins. PMID- 19544077 TI - A study of the evaporative deposition process: pipes and truncated transport dynamics. AB - We consider contact line deposition of an evaporating thin drop. Following Dupont's proposal (unpublished), we focus on transport dynamics truncated by a maximal concentration as the single deposition mechanism. The truncated transport process, formalized as the "pipe model", admits a characteristic shock front that has a robust functional form and depends only on local hydrodynamic properties. By applying the pipe model, we solve the density profile in different asymptotic regimes. In particular, we find that near the contact line the density profile follows a scaling law that is proportional to the square root of the concentration ratio defined as the initial solute volume concentration divided by the maximal concentration. The maximal deposit density occurs at about 2/3 of the total drying time for uniform evaporation and 1/2 for diffusion-controlled evaporation. Away from the contact line, areal density decays exponentially with the radial distance to the power of -3 for the uniform evaporation and -7 for the diffusion-controlled evaporation. PMID- 19544078 TI - AChR deficiency due to epsilon-subunit mutations: two common mutations in the Netherlands. AB - Congenital myasthenic syndromes are a clinically and genetically heterogeneous group of hereditary disorders affecting neuromuscular transmission. We have identified mutations within the acetylcholine receptor (AChR) epsilon-subunit gene underlying congenital myasthenic syndromes in nine patients (seven kinships) of Dutch origin. Previously reported mutations epsilon1369delG and epsilonR311Q were found to be common; epsilon1369delG was present on at least one allele in seven of the nine patients, and epsilonR311Q in six. Phenotypes ranged from relatively mild ptosis and external ophthalmoplegia to generalized myasthenia. The common occurrence of epsilonR311Q and epsilon1369delG suggests a possible founder for each of these mutations originating in North Western Europe, possibly in Holland. Knowledge of the ethnic or geographic origin within Europe of AChR deficiency patients can help in targeting genetic screening and it may be possible to provide a rapid genetic diagnosis for patients of Dutch origin by screening first for epsilonR311Q and epsilon1369delG. PMID- 19544079 TI - Reactivation of hepatitis B virus following systemic chemotherapy for malignant lymphoma. AB - Reactivation of hepatitis B virus (HBV) has been reported not only in HBsAg positive patients undergoing systemic chemotherapy, but also in a proportion of HBsAg-negative patients with HBc antibody and/or HBs antibody. Recently, rituximab-plus-steroid combination chemotherapy (R-CHOP, etc.) has been identified as a risk factor for HBV reactivation in HBsAg-negative patients with malignant lymphoma. Prophylaxis with antiviral drugs is essential for preventing HBV reactivation in HBsAg-positive patients, but there is little evidence on which to base the choice of drug or appropriate duration of prophylaxis. There are also few clinical data on HBsAg-negative patients and no established standard of care for such patients with HBV reactivation. Based on the limited number of previous reports, preemptive therapy, guided by serial HBV-DNA monitoring, is a reasonable strategy to prevent HBV reactivation in HBsAg-negative patients. However, clinical evidence alone is insufficient for determining optimal frequency of HBV-DNA monitoring during and after chemotherapy, or for determining when to stop preemptive therapy for HBV reactivation. Thus, well-designed clinical trials should be carried out to investigate the efficacy and safety of such preemptive therapy. Additionally, assessment of viral factors such as HBV genotypes and gene mutations may assist in the development of strategies to prevent the occurrence of severe hepatitis. In this review, we summarize the characteristics of HBV reactivation after systemic chemotherapy including rituximab, and propose a management strategy for malignant lymphoma patients suffering from HBV reactivation. PMID- 19544080 TI - Influence of the oxygen microenvironment on the proangiogenic potential of human endothelial colony forming cells. AB - Therapeutic angiogenesis is a promising strategy to promote the formation of new or collateral vessels for tissue regeneration and repair. Since changes in tissue oxygen concentrations are known to stimulate numerous cell functions, these studies have focused on the oxygen microenvironment and its role on the angiogenic potential of endothelial cells. We analyzed the proangiogenic potential of human endothelial colony-forming cells (hECFCs), a highly proliferative population of circulating endothelial progenitor cells, and compared outcomes to human dermal microvascular cells (HMVECs) under oxygen tensions ranging from 1% to 21% O2, representative of ischemic or healthy tissues and standard culture conditions. Compared to HMVECs, hECFCs (1) exhibited significantly greater proliferation in both ischemic conditions and ambient air; (2) demonstrated increased migration compared to HMVECs when exposed to chemotactic gradients in reduced oxygen; and (3) exhibited comparable or superior proangiogenic potential in reduced oxygen conditions when assessed using a vessel forming assay. These data demonstrate that the angiogenic potential of both endothelial populations is influenced by the local oxygen microenvironment. However, hECFCs exhibit a robust angiogenic potential in oxygen conditions representative of physiologic, ischemic, or ambient air conditions, and these findings suggest that hECFCs may be a superior cell source for use in cell-based approaches for the neovascularization of ischemic or engineered tissues. PMID- 19544081 TI - A highly selective, orally bioavailable, vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor has potent activity in vitro and in vivo. AB - Angiogenesis is a complex process that relies on a variety of growth factors and signaling pathways to stimulate endothelial cell responses and establish functional blood vessels. Signaling through the vascular endothelial growth factor (VEGF) receptors is an important mediator of angiogenesis, a hallmark of tumor growth and metastasis. Inhibition of signaling through VEGF has been clinically validated with FDA-approvals of bevacizumab, sorafenib, and suntinib. Our goal was to discover an orally available, selective VEGFR-2 inhibitor. A novel oxime, 1-{4-[6-amino-5-(methoxyimino-methyl)-pyrimidin-4-yloxy]-2-chloro phenyl}-3-ethyl-urea (JNJ-38158471), was identified as a potent and selective inhibitor of VEGFR-2. While JNJ-38158471 shares some structure features with sorafenib, unlike sorafenib, it lacks Raf kinase activity. JNJ-38158471 inhibits VEGFR-2 (IC50 = 40 nM) and closely related tyrosine kinases, Ret (180 nM) and Kit (500 nM); it has no significant activity (>1 microM) against VEGFR-1 and VEGFR-3. At nanomolar levels, it inhibits VEGF-stimulated autophosphorylation of VEGFR-2 in a whole cell assay and inhibits VEGF-dependent endothelial migration. Once daily oral dosing of JNJ-3815871 to nude mice bearing human A431, HCT116, and A375 tumors resulted in up to 90% tumor growth inhibition. Strikingly, after termination of JNJ-38158471 monotherapy-treatment of A375 xenografts, tumor growth delay was significantly prolonged up to 4 weeks. Anti-tumor efficacy correlated well with the observed dose concentrations (on a mg/kg basis) necessary to inhibit VEGF-induced corneal angiogenesis in C57BL/6J mice. In addition, the compound inhibited spontaneous polyp formation in the APC min-mouse model. These data demonstrate that JNJ-38158471 is a well tolerated, orally available, highly selective VEGFR-2 inhibitor that may have therapeutic benefit in human malignancies. PMID- 19544082 TI - Egg and whole-body amino acid profile of African bonytongue (Heterotis niloticus) with an estimation of their dietary indispensable amino acids requirements. AB - African bonytongue, Heterotis niloticus, is a river fish from the Central and West Africa basin. The species presents a great potential for fish farming and has been increasingly raised in Central and South Cameroon. The total amino acid and proximate composition of the whole body of egg, larva, juvenile, immature, and adult Heterotis were determined. Ash, moisture, whole-body protein, and lipid contents were significantly affected by size (P < 0.05). On the other hand, the amino acid composition of the whole-body tissue, when expressed as a percentage of dietary protein, was not significantly different among ontogenetic stages (ranging from 0.2 to 400 g mean body mass). The amino acid composition of the eggs was quite different to the one of whole-body tissue with lower levels of methionine, proline, and glycine, and higher levels of arginine, histidine, isoleucine, leucine, threonine, valine, serine, and alanine. The A/E ratios of adult Heterotis muscle tissue are similar to those obtained for other fish species, except for histidine and tryptophan. Based on whole-body or muscle tissue indispensable amino acids (IAA) to A/E ratios, the IAA requirement profiles for Heterotis (from larva to adult) were estimated and are similar to those of other omnivorous fish species, except for tryptophan and histidine. PMID- 19544083 TI - Simultaneous in vivo truncation of pectic side chains. AB - Despite the wide occurrence of pectin in nature only a few source materials have been used to produce commercial pectins. One of the reasons for this is that many plant species contain pectins with high levels of neutral sugar side chains or that are highly substituted with acetyl or other groups. These modifications often prevent gelation, which has been a major functional requirement of commercial pectins until recently. We have previously shown that modification of pectin is possible through heterologous expression of pectin degrading enzymes in planta. To test the effect of simultaneous modification of the two main neutral pectic side chains in pectic rhamnogalacturonan I (RGI), we constitutively expressed two different enzymes in Arabidopsis thaliana that would either modify the galactan or the arabinan side chains, or both side chains simultaneously. Our analysis showed that the simultaneous truncation of arabinan and galactan side chains is achievable and does not severely affect the growth of Arabidopsis thaliana. PMID- 19544084 TI - The effect of the Naked Neck genotype (Nana), feeding and outdoor rearing on growth and carcass characteristics of free range broilers in a hot climate. AB - Alternative poultry production with special reference to free range broilers has increased significantly since the nineties in many regions of the world. Numerous factors influence the productive performance of this type of broilers: genotype (namely the use of naked neck animals), feeding and access to an outdoor area. The aim of this paper is to study the influence of each of these factors on the productive performance of free range broilers under commercial rearing conditions. A total of 3200, day old chicks of both sexes from naked neck and normally feathered genotypes were used in this trial. After a joint initiation phase, animals were divided into four different treatments with the combination of two concentrates (high vs low energy content) and management (access to outside park or not). Experiment lasted a total of 12 weeks. Live weight date was recorded weekly and a samples of animals from the trial were sacrificed at the age of 8, 10 and 12 weeks, when carcass characteristics were determined. Besides sex, the only factor that seems to affect growth characteristics was genotype as naked neck animals had poorer growth rates than normally feathered. No effect was detected on carcass yields and percentages of carcass components for any of the variables. From the data presented in this trial the practises associated with free range production are of relative inconsequence to the technical animal production parameters and can only be justified by a pressing need to differentiate these products from standard poultry products in what concerns both welfare issues and meat characteristics. The results also indicate that genetic material from alternative poultry production in Europe can be a useful option in poultry production development projects in the tropics. PMID- 19544085 TI - Physiological response of dromedary camels to road transportation in relation to circulating levels of cortisol, thyroid hormones and some serum biochemical parameters. AB - Transportation is often considered as one of the main causes of stress raising considerable interest, both in economic and animal welfare terms. The objective of the current study was to determine physiological response of dromedary camels to road transportation in relation to circulating levels of cortisol, thyroid hormones and some serum biochemical factors during summer conditions. Ten Iranian dromedary camels, five males and five females, were selected for the study. The study was conducted on three consecutive days in August 2008. At first day, blood samples were collected at 08:30 A.M., 09:30 A.M. and 01:30 P.M. to determine any possible variation in individual measurements due to diurnal changes or as a result of food and water deprivation for 5 h. Travel commenced on day 2 at 08:30 A.M. for 5 h, with a total of about 300 km traveled. At second day, blood samples were collected immediately before loading, at 08:30 a.m., after 1 h transport, at 09:30 A.M., and on the end of transportation, after unloading, at 01:30 P.M. Final blood sample was taken 24 h after arrival. In the current study no significant difference was observed in any parameter between sexes at each sampling time. The data related to day before transport had no significant differences between different times except for values obtained for cortisol that at 01:30 P.M. showed a significant decrease in comparison with data at 08:30 and 09:30. Circulating cortisol, T(4), T(3) and fT(4) levels was significantly higher after transportation compared with pre-transport values and returned to basal values within 24 h after transport. Transportation had effects on metabolism as demonstrated by increase in serum concentrations of glucose, NEFA, and urea nitrogen. Serum concentrations of glucose, NEFA, and urea nitrogen returned to basal values in final bleeding at 24 h after transport termination. In the current study transportation had no significant effects on serum concentrations of fT(3), triglycerides, cholesterol, beta-hydroxybutyrate, albumin and total protein. Taken together, the results obtained for short road transportation of dromedary camels showed a strong physiological response and provide some biomarkers for stress detection in this species. Further research to validate these potential biomarkers is necessary. PMID- 19544086 TI - Causative agents of superficial mycoses in Istanbul, Turkey: retrospective study. AB - The aim of the present study was to determine the percentage of agents, which can give rise to superficial fungal infections in Istanbul, Turkey. Between 2000 and 2007, the clinical samples collected from 8,200 patients attending the outpatient Dermatology Clinic at Mihrimahsultan Medical Center were examined by direct microscopy and culture. Pathogen fungi were detected in 5,722 of the patients. Of the isolates were 4,218 (74%) dermatophytes, 1,196 (21%) Candida sp., 170 (3%) Malassezia furfur, and 138 (2%) Trichosporon sp. Among the dermatophytes, Trichophyton sp. was the most common isolate followed by Epidermophyton floccosum (243) and Microsporum sp. Among the Candida species, C. albicans (549) was also frequently found. Onychomycosis was the most prevalent type of infection, followed by tinea pedis, tinea cruris, tinea corporis, and tinea capitis. In conclusion, our study showed that the most common isolated agents from superficial infections were T. rubrum being Candida sp. the second most prevalent. PMID- 19544087 TI - Connexin43 interacts with Caveolin-3 in the heart. AB - Gap junctions (GJs), collections of multiple intercellular channels between neighboring cells, are specialized channels facilitating intercellular electrical and chemical communication. GJs are important for synchronizing coupling and coordinated contraction in the heart, and are crucial regulators of heart gene transcription, cardiac development, and protection of ischemic cardiomyocytes through second messenger communication. Identification of proteins that interact with Connexin43 (Cx43), the predominant protein in cardiac GJs, may contribute to the understanding of GJ functional regulation. Using a yeast two-hybrid system, we identified Caveolin-3 (Cav3) as a new Cx43-interacting protein. This interaction was confirmed by co-immunoprecipitation and co-localization experiments. CX43 interacts with Cav3, suggesting that Cav3 may participate in the functional regulation of GJs. PMID- 19544088 TI - Perceptions of the effectiveness of ethical guidelines: an international study of physicians. AB - The intent of ethics is to establish a set of standards that will provide a framework to modify, regulate, and possibly enhance moral behaviour. Eleven focus groups were conducted with physicians from six culturally distinct countries to explore their perception of formalized, written ethical guidelines (i.e., codes of ethics, credos, value and mission statements) that attempt to direct their ethical practice. Six themes emerged from the data: lack of awareness, no impact, marginal impact, other codes or value statements supersede, personal codes or values dictate, and ethical guidelines are useful. Overall, codes were valued only when they were congruent with existing personal morality. The findings suggest the need to re-evaluate the purpose, content, and delivery of codes for them to improve their function in promoting ethical conduct. PMID- 19544089 TI - PatientViewpoint: a website for patient-reported outcomes assessment. AB - PURPOSE: To develop a prototype website to collect patient-reported outcomes in outpatient clinical oncology and link the data with the electronic medical record (EMR). METHODS: A multidisciplinary Research Network, including experts in outcomes research, clinical oncology, nursing, social work, information technology, EMRs, behavioral science, decision science, clinical trials, law, and a cancer survivor, was formed to design the prototype website. The Research Network developed the initial website specifications, elicited feedback from patients (n = 20) and clinicians (n = 7), constructed the website, and conducted usability testing (n = 10). RESULTS: Clinicians reported that the website could improve clinical practice if it was not burdensome and were most interested in tracking change over time. Patients were interested in using the website because of the potential to facilitate communication with their clinicians. Patients emphasized the importance of short and simple surveys and a user-friendly interface. The PatientView-oint website was designed to meet these specifications. Usability testing suggested that patients had few problems accessing and using the site. CONCLUSIONS: Preliminary reports from clinicians and patients suggest that a website to collect PROs and link them with the EMR could help improve the quality of cancer care. Further pilot-testing will evaluate the use, usefulness, and acceptability of PatientViewpoint. PMID- 19544090 TI - When is a metric not a metric? Remarks on direct curve comparison in bioequivalence studies. AB - The majority of measures proposed to date for direct curve comparison in bioequivalence studies were investigated. These measures have often been called metrics, but in most cases this was incorrect in the mathematical sense. It was demonstrated, with a set of counter-examples, that the axioms of a metric are fulfilled only for the integral p-metric and some of its transforms. The Rescigno index and two other measures devised by Polli and McLean are the semi-metrics, lacking the triangle inequality, while others also lack symmetry. The use of the p-metric is therefore recommended, and statistical analysis is suggested as a point at which the scaling of differences might be carried out. PMID- 19544092 TI - Analysis of STAT5A/AvaI gene polymorphism in four Italian cattle breeds. AB - The STAT5A/AvaI polymorphism was investigated with PCR-RFLP in a sample of 339 cattle belonging to four breeds: Italian Friesian, Jersey, Italian Brown, and Podolica reared in south Italy. All three possible genotypes for the C/T polymorphism were identified. In these breeds, PCR-RFLP showed the predominance of the TT genotype in Italian Brown and Jersey cows; in Podolica and Italian Friesian CT is the most frequent genotype. The frequency of the T allele ranged from 0.55 to 0.81 in the analyzed populations. The distribution of genotypic and allelic frequencies at this locus was significantly different among the four populations based on a chi2 test (P < 0.001), suggesting that the molecular characteristics of the STAT5A gene could be significantly affected by the breed selection. Gene heterozygosity, gene homozygosity, effective allele number, fixation index, and polymorphism information content (PIC) were calculated. The observed heterozygosity, as well as the Ne and PIC values, indicates high genetic variability in the Podolica breed. Podolica could be considered an interesting reservoir of genetic diversity for a species under high selective pressure elsewhere. PMID- 19544093 TI - A 2.7-kb deletion in the human NLRP10 gene exon 2 occurred after the human chimpanzee divergence. AB - NLRP10 is a member of the NLRP protein family, which is involved in inflammation and apoptosis. Genome sequence comparisons revealed that a 2.7-kb deletion occurred in the human NLRP10 gene exon 2 after the divergence of humans and chimpanzees, resulting in replacement of the entire 3' untranslated region with the flanking LINE-1 element. The human NLRP10 protein lost 30 or more amino acids that are conserved in primates at its carboxy-terminus. The structural modification of the NLRP10 gene might have played a role in development or enhancement of human-specific traits during evolution. PMID- 19544091 TI - Using community-based participatory research to identify potential interventions to overcome barriers to adolescents' healthy eating and physical activity. AB - Using a community-based participatory research approach, we explored adolescent, parent, and community stakeholder perspectives on barriers to healthy eating and physical activity, and intervention ideas to address adolescent obesity. We conducted 14 adolescent focus groups (n = 119), 8 parent focus groups (n = 63), and 28 interviews with community members (i.e., local experts knowledgeable about youth nutrition and physical activity). Participants described ecological and psychosocial barriers in neighborhoods (e.g., lack of accessible nutritious food), in schools (e.g., poor quality of physical education), at home (e.g., sedentary lifestyle), and at the individual level (e.g., lack of nutrition knowledge). Participants proposed interventions such as nutrition classes for families, addition of healthy school food options that appeal to students, and non-competitive physical education activities. Participants supported health education delivered by students. Findings demonstrate that community-based participatory research is useful for revealing potentially feasible interventions that are acceptable to community members. PMID- 19544094 TI - The sustainability of evidence-based practices in routine mental health agencies. AB - The research presented here reports on sustainability of the practices within the National Implementing Evidence Based Practices Project for people with serious mental illness. Forty-nine sites completed the initial 2-year implementation phase and were the focus of our study. Our aims were to discern the number of sites that sustained practices 2 years after implementation, the reasons for sustaining or not sustaining, differences in characteristics between the two groups, and the extent and nature of practice adaptations. We used a mixed methods approach, based on a telephone survey that gathered qualitative and quantitative data from site representatives and others familiar with the sites and practices during the follow-up period. We found that 80% of sites sustained their practices for 2 years post-implementation, that sustainers differed from non-sustainers in several domains: financing, training, fidelity and agency leadership, and that most sites adapted practices moderately to meet state and local needs. PMID- 19544096 TI - Diabetes and apoptosis: liver. AB - The liver is a central regulator of glucose homeostasis and stores or releases glucose according to metabolic demands. In insulin resistant states or diabetes the dysregulation of hepatic glucose release contributes significantly to the pathophysiology of these conditions. Acute or chronic liver disease can aggravate insulin resistance and the physiological effects of insulin on hepatocytes are disturbed. Insulin resistance has also been recognized as an independent risk factor for the development of liver injury. In the healthy liver tissue homeostasis is achieved through cell turnover by apoptosis and dysregulation of the physiological process resulting in too much or too little cell death can have potentially devastating effects on liver tissue. The delineation of the signaling pathways that mediate apoptosis changed the paradigms of understanding of many liver diseases. These signaling events include cell surface based receptor-ligand systems and intracellular signaling pathways that are regulated through kinases on multiple levels. The dissection of these signaling pathways has shown that the regulators of apoptosis signaling events in hepatocytes can also modulate insulin signaling pathways and that mediators of insulin resistance in turn influence liver cell apoptosis. This review will summarize the potential crosstalk between apoptosis and insulin resistance signaling events and discuss the involved mediators. PMID- 19544095 TI - KLF4 suppresses HDACi induced caspase activation and the SAPK pathway by targeting p57(Kip2). AB - Kruppel-like factor 4 (KLF4) belongs to a family of evolutionarily conserved zinc finger-containing transcription factors. It has been shown to mediate self renewal and pluripotency, regulate adipogenesis and play a critical role in monocyte differentiation. KLF4 is also highly expressed in squamous cell carcinomas and in 70% of all primary human breast cancers, suggesting a putative role for KLF4 as being an oncogene and as an antiapoptotic factor. However, the mechanism of this regulation remains unclear. Here, we show that KLF4 is induced during histone deacetylase inhibitor treatment, and regulates the extrinsic apoptosis pathway by inhibiting caspase cleavage. In addition, KLF4 binds to the p57(Kip2) promoter and transcriptionally upregulates its expression, which in turn inhibits the stress activated protein kinase cascade and c-Jun phosphorylation. Our findings indicate that in cancer cells that express high levels of KLF4 may be refractory to HDACi treatment. Results of our study demonstrate an unexpected antiapoptotic function of KLF4, and suggest an important cell fate determinant following histone deacetylase inhibitor induced apoptosis. PMID- 19544097 TI - Safety profile of cefditoren. A pooled analysis of data from clinical trials in community-acquired respiratory tract infections. AB - INTRODUCTION: A high number of individuals in the population are exposed to antibiotics for the treatment of respiratory tract infections. It is important to review the adverse events profile related to antibiotic exposure during the clinical development of drugs that are or have been recently included in the therapeutic armamentarium. MATERIAL AND METHODS: Safety data from all 13 clinical trials of cefditoren on community acquired respiratory infections were reviewed. Safety population was defined as all randomized patients with at least one dose intake. Adverse events considered by investigators as related during antibiotic exposure were considered. RESULTS: The overall safety population consisted in 4,592 patients for cefditoren and 2,784 for comparators. Overall reported diarrhoea related to cefditoren administration was significantly higher (p < or = 0.001) than comparators (9.9% vs 6.9%) due to the significant difference in the pooled pharyngotonsillitis studies (8.3% vs 3.2%), while no significant differences in others pathologies were found, with 9.4% (with cefditoren) vs 10.3% (with comparators) in the case of community-acquired pneumonia (CAP). Dyspepsia and abdominal pain were reported as adverse events in < 2.7% patients regardless the treated disease. In females population lower related vaginosis rate was found in cefditoren vs comparators, mainly due to differences among patients treated for sinusitis (4.5% vs 8.1%) and CAP (2.3% vs 5.5%) although differences were not significant (p = 0.017 and p = 0.008, respectively). CONCLUSION: This study analysing reported adverse events from clinical trials showed an adverse events profile of cefditoren similar to those of standard antibiotics used in the treatment of respiratory tract infections. PMID- 19544098 TI - [Hydatid disease in the Department of Health #2 of the Valencian Community (Spain)]. AB - Hydatidosis is one of the most important zoonotic parasitic diseases of Mediterranean and South American countries where they constitute a significant health problem. Its diagnosis is realized essentially by image techniques and its treatment continues being eminently surgical. During the period 2000-2005, we reviewed clinical records of diagnosed and treated cases of hydatidosis carried out at the Department of Health 2 of the Valencia Community, Spain. Thirty three cases were evaluated of which we describe demographic, radiological, clinical, analytic, therapeutical and evolutionary data. We discuss the peculiarities found in them: thirty two cases were of hepatic location, essentially right, 48.5% unique cysts, five casual finding, 88.87% were diagnosed by ultrasonographic technique and in 2/3 of the cases the serology was positive. In addition, 52.17% of the twenty three cases treated surgically cured without recurrence and a 15.15% recurred without subsequent healing. Studies will require long-term monitoring of patients evolution. PMID- 19544099 TI - [Diagnosis of urethritis in men. A 3-year review]. AB - OBJECTIVES: The aim of this study is to know the prevalence and tendency of microorganisms producing urethritis, in men, in the City Centre of Madrid. METHODS: Cross-sectional study. The urethral samples of 1.248 men were analyzed, for 3 years. The samples were studied for: GRAM stain, when secretion exists; culture in habitual plates; detection of C. trachomatis, U. urealyticum and M. hominis, when there was suspicious, study of T. vaginalis and when suspicious injuries exist, study of virus Herpes simplex. RESULTS: The percentage of positive samples was 22.60%. The isolated microorganisms were: U. urealyticum 7.61%, N. gonorrhoeae 6.33%, C. trachomatis 4.81%, M. hominis 0.24%, H. parainfluenzae 1.76%, H. influenzae 1.12%, Candida spp 0.48%, S. pyogenes 0.16% and Herpes virus simplex (2) 0.08%. Two or more microorganisms were isolated in 1.68%. The percentage of positive samples in 2003 was 17.41% and N. gonorrhoeae the most frequent microorganism (6.22%). In 2004 was 25.57% and the most frequent U. urealyticum (10.18%). In 2005 the 24.50% of the samples were positive and U. urealyticum the most frequent (7.92%). The 79.41% of N. gonorrhoeae were susceptible to all antibiotics tested. It is not found resistance to ceftriaxone, claritromicine and amoxicilline/clavulanic acid. The 11.76% were betalactamase- producing. The 26.47% of Haemophilus spp. were betalactamase- producing and all strains were susceptible to cefotaxime. CONCLUSIONS: The isolated microorganisms most frequently were: U. urealyticum, N. gonorrhoeae and C. trachomatis. There is an increase of 7% of prevalence between the years 2003 and 2005. Ceftriaxone, claritromicine and amoxicilline/clavulanic acid were susceptible to all the strains studied and cefotaxime to all Haemophilus spp. PMID- 19544100 TI - [High percentage of clarithromycin and metronidazole resistance in Helicobacter pylori clinical isolates obtained from Spanish children]. AB - OBJECTIVE: To determine the primary and secondary resistance to several antimicrobial agents in Spanish Helicobacter pylori clinical isolates obtained from paediatric patients from January 2002 to June 2006. METHODS: Samples were collected from gastric biopsies of symptomatic paediatric patients and H. pylori cultured according to standard microbiological procedures. Resistance was determined by E-test. Strains were considered resistant if minimal inhibitory concentration (MIC) > or = 2 mg/l for amoxycillin, > or = 4 mg/l for tetracycline, > or = 8 mg/l for metronidazole, > or = 1 mg/l for clarithromycin, MIC > or = 4 mg/l for ciprofloxacin, MIC > or = 32 mg/l for rifampicin and intermediate if MIC = 0.5 mg/l for clarithromycin, and MIC = 2 mg/l for ciprofloxacin. RESULTS: A total of 101 patients were included: 38 males and 63 females (sex ratio M/F: 0.6). Average age was 10 years (range: 4-18 years). All strains were susceptible to amoxycillin, tetracycline and rifampicin, 35.7% were resistant to metronidazole, 54.6% to clarithromycin and 1.8% to ciprofloxacin. 2.0% were intermediate to clarithromycin and 1.8% to ciprofloxacin. Double resistance to metronidazole and clarithromycin rated at 17.2%. Thirty-five patients (34.7%) had a history of treatment failure, and were considered as secondary H. pylori. Primary resistance rates to metronidazole and clarithromycin were 32.8% and 49.2%, respectively, and secondary resistance rates were 41.2% and 70.6%, respectively. CONCLUSIONS: Resistance to clarithromycin (56.6%) was higher than to metronidazole (35.7%) in the H. pylori strains studied. Clarithromycin resistance was very high even in strains from paediatric patients not previously treated for H. pylori infection. PMID- 19544101 TI - [Antiparasitic drugs. Review of the useful drugs in the treatment of classic and emergent parasitic diseases]. AB - INTRODUCTION: In the 10 last years, the increase of international travels and immigration from low income countries to Spain was related with an increased of prevalence of parasitic diseases. SOURCE: Critical review of the literature. STRUCTURE: Firstly, several general considerations were made on the antiparasitic drugs revised in this paper. Chemical structures and mechanisms of action of the main drugs with antiparasitic effect were considered in the second part of the review. Further, antiparasitic spectrum of selected drugs, main pharmacokinetical characteristics, usual posology, possible side effects and contraindications were detailed. Finally, some practical aspects, such as interactions and the methods for practical obtention of these drugs are indicated. This information is relevant because in Spain many anti-parasitic drugs may be obtained using non conventional methods. CONCLUSION: In Spain, the increase of parasitic diseases necessitates an update on antiparasitics drugs for their treatment. PMID- 19544103 TI - Lung function, sociodemographic characteristics, and psychological reaction to transplant associated with chronic stress among lung recipients. AB - Chronic stress is a well-known consequence of somatic diseases. In this study, we investigated whether physical, sociodemographic, or transplant-related psychological factors were associated with the patient's chronic stress level. A cross-sectional study enrolling 76 patients measured chronic stress (Screening Scale, Screening Subscale of Chronic Stress of the Trier Inventory for the Assessment of Chronic Stress) and the emotional effects of the transplant (Transplant Effects Questionnaire), as well as physical and sociodemographic conditions (lung function, bronchiolitis obliterans syndrome, working status, and parenting). Chronic stress after a lung transplant was significantly lower than in a normal community sample. In the multiple regression analysis, worries concerning the transplant were significantly associated with the patient's chronic stress, but not with physical or sociodemographic parameters, nor with interactions between physical and psychological parameters. These results underscore the importance of transplant-related worries, regardless of the patient's current state of health. PMID- 19544104 TI - Metrics for comparing three word-based software programs used for augmentative and alternative communication. AB - The purpose of this investigation was to conduct an analysis of a set of metrics developed for comparing features of software used in speech generating devices (SGD) for augmentative and alternative communication (AAC). These metrics - measures of speed, efficiency, and accuracy - were employed during a sentence reconstruction task. Twenty two participants without disabilities reconstructed nine sentences using each of three SGD software programs, counterbalanced to remove order effects. Results revealed differences in the metrics across the software programs that were supported by a social validation survey completed by the participants. These results provide benchmark data about relative speed, accuracy, and efficiency of these software programs in structured use, and can inform professionals in matching potential AAC users with specific features of word-based software. PMID- 19544102 TI - [Tinidazole: a classical anaerobical drug with multiple potential uses nowadays]. AB - Tinidazole is a 5-nitroimidazole active in vitro against a wide variety of anaerobic bacteria and protozoa. Tinidazole is an effective treatment against anaerobic microorganisms based on its pharmacokinetic characteristics (C(max) 51 microg/ml, t(1/2) 12.5 h) and its excellent in vitro activity. Its long half-life allows once a day regimens. Tinidazole is as effective as metronidazole in the treatment of infections caused by T. vaginalis, giardiasis and amebiasis and bacterial vaginosis, malaria, odontogenic infections, anaerobic bacterial infections (pelvic inflammatory disease, diabetic foot), surgical prophylaxis (abdominal and hysterectomy) and Helicobacter pylori eradication. Tinidazole was recently approved by the Food and Drug Administration (FDA) for the treatment of infections caused by Trichomonas vaginalis, Entamoeba histolytica and Giardia lamblia. PMID- 19544105 TI - Reliability of speech generating devices: a 5-year review. AB - Individuals who use augmentative and alternative communication (AAC) depend on technology to meet their daily needs and form relationships. Speech generating devices (SGDs) are integral components of communication systems. Reliability of SGDs is critical for effective use in everyday life. This study examined the reliability of new SGDs and found that mean time to first failure was 42.7 (SD = 41.2) weeks and at least 40% required repairs within the first year of use. The components that most frequently broke down were touch screens, wiring, main boards, batteries, memory cards, and AC adaptors. The costs of repairing SGDs were analyzed. The clinical implications of device breakdown are identified for key stakeholders, including clients, families, service providers, funding agencies, and manufacturers. PMID- 19544108 TI - Chromosome 3p microsatellite allelotyping in neuroblastoma: a report on the technical hurdles. AB - Pinpointing critical regions of recurrent loss may help localize tumor suppressor genes. To determine the regions of loss on chromosome 3p in neuroblastoma, we performed loss of heterozygosity analysis using 16 microsatellite markers in a series of 65 primary tumors and 29 neuroblastoma cell lines. In this study, we report the results and discuss the technical hurdles that we encountered during data generation and interpretation that are of relevance for current studies or tests employing microsatellites. To provide functional support for the implication of 3p tumor suppressor genes in this childhood malignancy, we performed a microcell-mediated chromosome 3 transfer in neuroblastoma cells. PMID- 19544110 TI - Activated PKCalpha/ERK1/2 signaling inhibits tamoxifen-induced apoptosis in C6 cells. AB - Data have shown that tamoxifen (TAM) can be used to treat not only breast cancer with positive estrogen receptor (ER), but also negative ER including human glioma. However, the molecular mechanism of this drug against different kinds of cancers remains to be elucidated. In this study, we provided the evidence that PKCalpha-ERK1/2 signaling pathway plays a negative role in TAM-induced C6 cell apoptosis, and a combined utilization of TAM with inhibitors of PKCalpha or ERK1/2 could enhance the effectiveness of TAM on inhibiting tumor growth. PMID- 19544111 TI - Association between MGMT promoter hypermethylation and p53 mutation in glioblastoma. AB - O(6)-methylguanine-DNA methyltransferase (MGMT) is a DNA repair enzyme that removes alkyl groups from the O(6) position of guanine. MGMT is transcriptionally silenced by promoter hypermethylation in several human neoplasia. We used methylation-specific PCR (MSP) to analyze the MGMT promoter methylation status of 50 glioblastoma tumors. Hypermethylation was detected in 24 of 50 (48%) samples. We also analyzed mutant p53 expression by immunohistochemical analysis of glioblastoma tissue samples. A significant association was found between MGMT methylation and p53 mutation status (p< .05). These results suggested that epigenetic inactivation of MGMT plays an important role in the survival of glioblastoma patients and this inactivated gene involved in p53 mutation. PMID- 19544106 TI - The relationship of oxidative stress and cholesterol with dipping status before and after aerobic exercise training. AB - OBJECTIVE: The purpose of this study was to examine the effects of aerobic exercise training (AEXT) on dipping status in pre-hypertensive and stage-1 hypertensive individuals. A secondary purpose was to evaluate whether AEXT alters oxidative stress and endothelial biomarkers correlated to dipping status. METHODS: Twenty-three subjects underwent 24-h ambulatory blood pressure monitoring at baseline and after 6 months of AEXT. AEXT consisted of training at 70% VO(2max) 3 days/week for 6 months. Total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein (LDL)-cholesterol, oxidized LDL (ox-LDL), triglycerides, urinary and plasma nitric oxide end-products, superoxide dismutase and 8-iso-PGF(2alpha) were measured before and after AEXT. Statistically, ANOVA and linear regression were used. RESULTS: Before and after AEXT, there were no significant differences between dippers and non-dippers in any of the biomarkers except for total cholesterol following AEXT. In a sub analysis following AEXT, 14 subjects retained their original dipping status, five subjects changed from dippers to non-dippers and four subjects changed from non dippers to dippers. Significant differences existed between these groups in changes in total and LDL-cholesterol, ox-LDL, 8-iso-PGF(2alpha) and % Dip. CONCLUSIONS: Changes in cholesterol levels but not oxidative stress or endothelial biomarkers were related to changes in BP variables following AEXT in dippers and non-dippers. PMID- 19544113 TI - Very low birth weight and attention deficit/hyperactivity disorder. AB - It is well recognized that reaction time performance of term-born children with a normal birth weight (NBW > 2500 g) who fulfill the DSM-IV criteria for attention deficit/hyperactivity disorder (ADHD) in the primary school age is sensitive for the presentation rate of stimuli. They have been found to perform more poorly in conditions of relatively slow event rates as compared with fast event rates. The purpose of the current study was to explore whether reaction time performance of children with very low birth weight (VLBW < 1500 g) with or without ADHD showed the same sensitivity for the factor presentation rate of stimuli compared to children with a normal birth weight plus ADHD. To this end, reaction time performance of four groups of children was compared on a Go/No-Go test with a fast presentation rate of 1 second and a slow presentation rate of 6 seconds. Groups were: children with VLBW plus ADHD, children with VLBW only, children born full term with normal birth weight (NBW >2500 g) plus ADHD, and children born full term without ADHD (the control group). Findings indicated that, compared to the non-ADHD groups, the groups with ADHD (NBW and VLBW) showed a more dramatic decline in their reaction time performance in the slow condition: a state regulation deficit. In addition, both groups showed a response inhibition deficit. No difference was found in reaction time performance between the groups of children with VLBW only and the control group. PMID- 19544114 TI - Difference between palpation and optoelectronics recording of scapular motion. AB - The aim of this study is to determine the errors of scapular localisation due to skin relative to bone motion with an optoelectronic tracking system. We compared three-dimensional (3D) scapular positions obtained with skin markers to those obtained through palpation of three scapular anatomical landmarks. The scapular kinematics of nine subjects were collected. Static positions of the scapula were recorded with the right arm elevated at 0 degrees, 40 degrees, 80 degrees, 120 degrees and 160 degrees in the sagittal plane. Palpation and subsequent digitization of anatomical landmarks on scapula and thorax were done at the same positions. Scapular 3D orientation was also computed during 10 repeated movements of arm elevation between 0 degrees and 180 degrees. Significant differences in scapular kinematics were seen between static positions and palpation when considering anterior/posterior tilt and upward/downward rotation at angles over 120 degrees of humeral elevation and only at 120 degrees for internal/external rotation. There was no significant difference between positions computed during static positions and during the movement for the three scapular orientations. A rotation correction model is presented in order to reduce the errors between static position and palpation measurement. PMID- 19544115 TI - Sex after ART: sexual partnerships established by HIV-infected persons taking anti-retroviral therapy in Eastern Uganda. AB - This paper explores the social contexts that influence the formation and nature of sexual partnerships among people on anti-retroviral therapy (ART). We draw on the findings of a qualitative, longitudinal study of 70 people (36 women and 34 men) who have been participating in a home-based ART programme for over three years in Eastern Uganda. Since initiating ART, 32 (18 men and 14 women) participants reported having had a new partner. Five participants (4 men and 1 woman) renewed relationships with spouses with whom they had been prior to starting ART. Overall, 37 of the 70 participants had had a sexual partner after starting ART. Companionship, material support, social and cultural norms, as well as a desire for sex and children, are drivers of new relationships. The opportunity that ART brings for people to get on with their lives brings with it a reinstatement into a social world that places a value on marriage and child bearing. The sexual rights of those living with HIV and on ART need to be taken seriously and safer sex facilitated. PMID- 19544117 TI - Leptin concentrations in the peritoneal fluid of women with ovarian endometriosis are different according to the presence of a 'deep' or 'superficial' ovarian disease. AB - Some studies have suggested a possible role of leptin, an active cytokine produced by adipocytes, in the pathogenesis of pelvic endometriosis. The present study was designed to assess leptin levels in the peritoneal fluid (PF) of women with the 'deep' or 'superficial' types of ovarian endometriosis. Twenty-seven women with a single ovarian endometrioma having a mean diameter between 3 and 5 cm were included in the study. Patients were divided into two groups according to the type of ovarian endometriosis: Group A (n = 11) consisted of women with 'superficial' endometriomas located at the ovarian surface; Group B (n = 16) included patients with 'deep' intra-ovarian endometriomas. Women undergoing laparoscopy for unexplained infertility and not affected by pelvic and/or ovarian endometriosis were considered as controls (Group C, n = 10). Patients with an ovarian endometrioma had significantly increased PF leptin concentrations than endometriosis-free controls (Groups A and B vs. Group C, p < 0.01). Patients with 'superficial' endometriomas had significantly higher PF leptin levels compared with patients with 'deep' endometriomas (Group A vs. B, p < 0.01). This difference remained significant after correction for the BMI; moreover, a positive correlation between PF leptin and BMI was observed in Groups B and C, but not in women with 'superficial' endometrioma (Group A). Our observations suggest that: (a) leptin could play an active role in promoting the development of 'superficial' ovarian endometriomas and (b) 'superficial' and 'deep' ovarian endometriomas could have a different pathogenesis. PMID- 19544116 TI - Computerized analysis of heart rate in fetuses from mothers under levothyroxin treatment. AB - OBJECTIVE: To investigate fetal heart rate (FHR) of fetuses whose mothers are under levothyroxin treatment for chronic hypothyroidism. STUDY DESIGN: Sixty women under chronic therapy with levothyroxin and 180 controls at 37-39 weeks' gestation were studied by Sonycaid Sistem 8002(R) computerized cardiotocography (cCTG) for 30 min. cCTG parameters were expressed as mean and SD and the differences tested for statistics by Student t-test. Furthermore, cCTG parameters were related to levothyroxin dose by regression analysis. Significance was assessed at p < 0.05. RESULTS: Computerized cardiotocographic tracings of fetuses from mothers under levothyroxin treatment revealed: significant reduction of baseline FHR (130.1 +/- 9.47 vs. 134.9 +/- 4.68 bpm); increased number of FHR decelerations greater than 20 bpm (0.2 +/- 0.41 vs. 0.05 +/- 0.22); reduction of body movements per hour (6.68 +/- 11.72 vs. 10.65 +/- 11.74); and increased uterine contraction peaks (5.15 +/- 4.69 vs. 2.7 +/- 2.57). Those fetuses also showed significantly reduced neonatal weight (2668.2 +/- 766.65 vs. 3215.44 + 523.88 g) and lower 1-min Apgar score (8.6 +/- 0.95 vs. 9.3 +/- 1.11). Regression analysis showed a significant correlation between levothyroxin dose and baseline FHR (r = 0.60; p < 0.0001) and fetal body movements per hour (r = 0.52; p < 0.0001), and an inverse relationship with uterine contraction peaks (r = -0.35; p < 0.006), whilst no correlation was found with the number of FHR decelerations greater than 20 bpm. CONCLUSIONS: Maternal hypothyroidism and levothyroxin treatment influence FHR and cCTG is a sensible tool to reveal that influence. PMID- 19544118 TI - The contributions of resistin and adiponectin gene single nucleotide polymorphisms to the genetic risk for polycystic ovary syndrome in a Japanese population. AB - Polycystic ovary syndrome (PCOS) is a heterogeneous group of disorders that occur fairly commonly in women of reproductive age and are characterized by a variety of clinical manifestations, including insulin resistance that is independent of obesity. Recent studies suggest that altered adipocytokine gene expression is closely associated with insulin resistance and that single nucleotide polymorphisms (SNPs) modulate the expression and/or function of these genes, thereby affecting insulin sensitivity. With that in mind, we investigated whether SNPs at position -420 of the resistin gene (RETN) and/or -11377 of the adiponectin gene (ADIPOQ) modulate the susceptibility to PCOS. We evaluated the genotypes of 117 women with PCOS and 380 healthy fertile controls and measured the index of insulin resistance and hormonal profiles in the PCOS women. The RETN 420G/G homozygous variant genotype occurred significantly more frequently among the PCOS group than among the control group (15.4% vs. 8.4%, p = 0.035). PCOS women with the RETN-420G/G genotype also showed significantly higher BMIs and greater insulin resistance than those with RETN-420 C/C or C/G genotypes. The ADIPOQ SNP at -11377 showed no association with PCOS. We conclude that the RETN G/G at -420 genotype is associated with PCOS in Japanese women. PMID- 19544119 TI - Large-for-gestational-age infants of type 1 diabetic mothers: an effect of preprandial hyperglycemia? AB - OBJECTIVE: To determine how the frequency, timing and magnitude of hyperglycemia are associated with large-for-gestational-age (LGA) infants in pregnancies complicated by type 1 diabetes. METHODS: Charts from pregnant women with type 1 diabetes (n = 70) were reviewed. Indices of maternal glycemic control were determined for seven gestational periods (weeks 7-10, 11-15, 16-19, 20-24, 25-28, 29-32 and 33-38) and compared between women who delivered LGA infants and appropriate-for-gestational-age (AGA) infants. RESULTS: Of the 70 pregnancies, 57% of the infants were LGA (4.3 +/- 0.4 kg) and 43% were AGA (3.2 +/- 0.4 kg). Total maternal weight gain and rate of weight gain were significantly higher in mothers with LGA infants. The glycemic variables associated with an LGA infant were percentage of preprandial values above target for weeks 11-15, 25-28 and 29 32, and percentage of all values above target for weeks 33-38. For the entire pregnancy, the strongest predictors of an LGA infant were percentage of preprandial blood glucose values above target during weeks 29-32 and maternal weight gain. CONCLUSIONS: In pregnant women with type 1 diabetes, frequent episodes of preprandial hyperglycemia in the third trimester significantly impact the development of LGA infants. PMID- 19544120 TI - Hormonal replacement therapy in women with Turner's syndrome in Poland: Analysis of 176 cases. AB - AIM: Turner syndrome (TS) is one of the forms of gonadal malfunction. The study aims at the analysis of hypophysis-gonad axis (HGA) of women with TS who use and do not use hormonal replacement therapy (HRT). METHOD: One hundred and seventy six Poles with TS were investigated in the years 1995-2004. The information about the application of HRT was given during the interview. The HGA was examined by the estimation of gonadotropin (FSH, LH) and 17beta-estradiol (E(2)) levels. RESULTS: HRT was administered to 89% of women with TS at some point of their life. However, at the time of writing this report only 54% of them have been taking this drug. The variety of hormonal preparations used by the patients was great. In women with TS using the HRT, in contradiction to women with TS who do not use it, lower concentrations of FSH (32.1 +/- 22.1 vs. 44.2 +/- 23.3 IU/l) and LH (20.8 +/- 17.5 vs. 26.6 +/- 18.1 IU/l) as well as higher level of E(2) (135.5 +/- 147.9 vs. 89.9 +/- 100.6 pmol/l) were observed. The negative correlation between E(2) and FSH levels was not observed in women with TS using HRT, despite the elevation of 17beta-estradiol levels and the reduction of gonadotropin concentration. CONCLUSIONS: (1) Large percentage of women with TS does not use HRT. (2) Women with TS, who receive HRT, use this method of treatment insufficiently. PMID- 19544121 TI - Poor ovarian response in patients younger than 35 years: is it also a qualitative decline in ovarian function? AB - OBJECTIVE: To investigate whether poor response to controlled ovarian stimulation (COS) is due to a qualitative decline in ovarian function. METHODS: This retrospective cohort study included 436 patients younger than 35-years old, undergoing COS for intracytoplasmic sperm injection (ICSI). Patients with four or fewer MII oocytes after COS (poor-responder group, PR, n = 52) were age-matched with normoresponder patients (NR, n = 364). RESULTS: Although similar duration of stimulation (10.5 +/- 0.4 and 9.3 +/- 0.8 days; p = 0.1358), increased doses of gonadotrophins (2510 +/- 865 and 2253 +/- 572 IU; p = 0.0061) were used in the PR. The results show a increased chance of cycle ending of PR (PR: 26.9% and NR: 3.1%; p < 0.0001). Although the lower total number of oocytes retrieved (2.4 +/- 1.4 and 16.2 +/- 9.3; p < 0.0001), equal rate of fertilization (70.2% and 72.0%, p = 0.1190) and high quality embryos were obtained (50.0% and 45.2%; p = 0.4895), resulting in similar implantation (14.5% and 19.7%; p = 0.2246) and abortion (10.0% and 15.4%; p = 1.00) rates, respectively. A trend towards increased pregnancy rate per embryo transfer in NR group was noted (PR: 26.3% and NR: 42.2%; p = 0.0818). CONCLUSIONS: Low ovarian response could be associated mainly with a quantitative rather than a qualitative decline in ovarian function. Therefore, even if the ovarian response to stimulation is low, patients aged < or =35 years should process to oocyte retrieval. PMID- 19544124 TI - Introducing a special issue on the mental health of the girl child. PMID- 19544123 TI - Preimplantation genetic diagnosis for the prevention of sickle cell disease: current trends and barriers to uptake in a London teaching hospital. AB - INTRODUCTION: Sickle cell disease (SCD) is a clinically significant hemoglobinopathy with increasing global incidence. We describe our experience of using pre-implantation genetic diagnosis (PGD) for the prevention of SCD at a tertiary referral centre in London. METHODS: Between January 2002 and December 2007, of 78 at-risk couples referred for PGD treatment, 12 couples (15%) underwent 16 PGD cycles for the prevention of SCD. RESULTS. The live birth rate was 13% per initiated cycle, 18% per embryo transfer and 17% per couple. CONCLUSIONS: Although PGD for prevention of the birth of a child affected by SCD is a viable treatment option for couples at risk of having an affected child, potential barriers to uptake of this service need to be fully addressed to ensure its availability to all couples seeking to avoid having a child affected with SCD. PMID- 19544125 TI - Uprooted and displaced: a critical narrative study of homeless, Aboriginal, and newcomer girls in Canada. AB - Uprooting and displacement are a common part of everyday life for millions of girls and young women throughout the world. While much of the discourse has centered on movement from one country to another, uprooting and displacement are also a reality for many within Canada. Notably, a growing population of homeless girls and Aboriginal girls also have experienced uprooting and dislocation from home, community, and in some cases, family. For many of these girls, multiple forms of individual and systemic violence are central features of their lives. The primary purpose of this critical narrative study is to examine how uprooting and displacement have shaped mental health among three groups: (1) newcomers to Canada (immigrant and refugee girls); (2) homeless girls; and (3) Aboriginal girls. In-depth narrative interviews were conducted with 19 girls in Southwestern Ontario. Narrative themes revealed that although there is much diversity within and between these groups, uprooting and displacement create social boundaries and profound experiences of disconnections in relationships. Barriers to re/establishing connections generate dangerous spaces within interlocking systems of oppression. However, in negotiating new spaces, there is the potential for the forming and re-forming of alliances where sources of support hold the promise of hope. It is within these spaces of hope and pathways of engagement where connections offer a renewed sense of belonging and well-being. The findings highlight the relevance of the construct of uprootedness in girls' lives, provide beginning directions for the design of gender-specific and culturally meaningful interventions, and comprise a substantial contribution to the growing body of research related to girls and young women. PMID- 19544126 TI - Mental health of the African girl child: challenges for nurses. AB - In this paper, the ecological systems theory of child development is used as the framework for exploring research on the development of African children, especially girl children. The special problems faced by children and the challenges this raises for nurses also are described in this paper. PMID- 19544128 TI - Childhood maltreatment among community dwelling adult Latinas. AB - There is a lack of research about childhood maltreatment among adult Latinas in the United States. The aim of this secondary analysis was to explore the occurrence and co-occurrence of maltreatment during childhood in a community dwelling sample of adult Latinas. Participants were 62 women ages 18-45 who read and spoke English, attended work or school on a regular basis, and had no health or functional interferences with daily living. On average, participants were 29 years of age (M = 29.08, SD = 9.30), high school graduates (n = 36, 58.1%), married (n = 36, 58.1%), and employed (n = 27, 43.5%). Over three quarters of participants (n = 49, 79.0%) reported at least one form of childhood maltreatment: emotional abuse (n = 24, 38.7%), physical abuse (n = 29, 46.8%), sexual abuse (n = 29, 46.8%), emotional neglect (n = 27, 43.5%), and physical neglect (n = 23, 37.1%). Maltreatment severity increased with the number of maltreatment forms. Women sexually abused during childhood were more likely than women without this history to be employed and high school graduates. These Latinas may have developed more adaptive coping mechanisms than other Latinas. PMID- 19544127 TI - Attitudes and beliefs concerning sexual relationships among minority adolescent women. AB - Learning about one's sexuality is an important developmental task for the adolescent and has implications for a lifelong process of becoming a sexual being and reaching sexual maturity. Learning about intimacy in relationship with others is another important developmental task for the adolescent. The occurrence of interpersonal violence, particularly sexual, physical, or psychological violence, disrupts the development of intimacy which informs the process for becoming a sexual being. Society can assist adolescent females experiencing interpersonal violence to understand their sexual nature and make informed sexual decisions throughout their lifetime. This assistance requires a contextual understanding of the relationship between adolescent sexual development and the effects of interpersonal violence on this process. This study presents the groundwork needed to understand the contextual meanings of sexual relationships of adolescent females with a history of interpersonal violence and discusses the evolution of these adolescents' attitudes and beliefs for sexual health promotion. PMID- 19544129 TI - Barriers to depression treatment in low-income, unmarried, adolescent mothers in a southern, urban area of the United States. AB - This study explored barriers to depression treatment in low-income, unmarried, adolescent mothers in a southern, urban area of the United States. The authors utilized a phenomenological approach and focus group methodology. Participants (n = 9) were enrolled in a teen parent program, an option of the public school system. The metaphor of a merry-go-round emerged from the data and represented the ups and downs that the adolescent mothers experience as they struggle to adjust to the role of mother. Their knowledge of postpartum depression and depression treatment occurred in the context of their demographics and their desire to create a family for their baby, their fears, and surprise at the reality of mothering. Childbirth education for adolescent mothers should include information on depression and the process of depression treatment. PMID- 19544130 TI - Sexual abuse and violence among adolescent girls in Botswana: a mental health perspective. AB - The presence of sexual abuse among societies in Botswana is a phenomenon whose occurrence is usually denied albeit the police report on it and legal frameworks have been established to combat it. Several factors influence the concealment of sexual abuse among adolescent girls, which includes cultural factors and social status of the perpetrators. This paper espouses the concept of sexual abuse among adolescent girls, the existence of the problem, its magnitude, the factors that increase vulnerability to violence and abuse, and how these factors intersect with HIV and AIDS. Two case studies using a discovery method were used to explore the phenomenon under the study. The findings of the study indicated that sexual abuse and violence have profound mental health consequences including guilt, anxiety, depression and anger. Future research is suggested to explore this problem on a wider scale and develop interventions that can assist victims and perpetrators to cope with the situation. PMID- 19544132 TI - Interoperability--making information systems work together. PMID- 19544131 TI - The social context of sexual health and sexual risk for urban adolescent girls in the United States. AB - Sexually transmitted infections including HIV and teenage pregnancy have resulted in considerable morbidity and mortality among girls in the United States. There is a need to further strengthen prevention efforts against these persistent epidemics. In order to promote girls' sexual health and most effectively reduce sexual risk, it is important to understand the social factors that influence the development of a girl's sexuality. The purpose of this study was to begin to fill a void in the literature by exploring girls' perspectives about the social context in which they learn about sex, sexuality, and relationships. Coding and content analysis was used to identify patterns and themes in 33 individual interviews with African American and Euro-American girls. Participants identified family, friends/peers, partners, school, and the media as the most common sources for learning about sexual health. Girls sought out different types of information from each source. Many girls experienced conflicting messages about their sexual health and struggled to integrate the disparate cultural references to sex, sexuality, and relationships that emerged from these different spheres of social life. Girls often had to navigate the journey of their sexual development with little room for reflection about their own thoughts, feelings, desires, and decisions. Health care providers, especially those in mental health, are in an optimal position to promote girls' physical, developmental, and emotional sexual health. PMID- 19544133 TI - Empathy in frontotemporal dementia and Alzheimer's disease. AB - Using naturalistic stimuli, we assessed the ability to infer what other people are feeling in three groups of participants: healthy elderly adults, patients suffering from the behavioral variant of frontotemporal dementia (FTD-b), and patients suffering from Alzheimer's disease (AD). After watching videotaped interviews of everyday people (nonactors) discussing an emotionally relevant event in their lives, participants answered questions regarding the interviewee's feelings. Both patient groups inferred emotions as accurately as the healthy elderly, provided the emotions were displayed unambiguously and consistently across the interview. However, when the displayed emotions became more variable and ambiguous, performance in both patient groups became impaired relative to healthy elderly participants. The similar profile across the two clinical groups despite their differences in social skills suggests that nonsocial cognitive processes affected in dementia may be an important factor in drawing inferences about other people's feelings. PMID- 19544134 TI - Gaze cueing of attention in schizophrenia: individual differences in neuropsychological functioning and symptoms. AB - Inhibition of return (IOR) represents a well-known mechanism of human perception that biases attentional orienting to novel locations in the environment. Behaviorally, IOR reflects slower reaction time (RT) to stimuli presented in previously cued locations. In this study, we examined within patients with schizophrenia this inhibitory aftereffect using two different cue types--eye gaze and standard peripheral cues. Results indicated that patients showed evidence of IOR, as reflected in a 3.2% slowing in RT to previously peripherally cued locations. However, for eye gaze, patients failed to show evidence of IOR and instead had 1.7% faster RT to targets presented following delay in locations that had been previously cued. This inhibitory failure correlated strongly with reduced neuropsychological performance and global symptoms ratings of attention and bizarre behavior. Reduced inhibitory aftereffect in RT for eye-gaze cues may reflect disease-related abnormalities in social attention. PMID- 19544135 TI - Occupational exposure to benzene at the ExxonMobil refinery at Baton Rouge, Louisiana (1977-2005). AB - Because crude oil contains up to 3% benzene and there is an association between high chronic exposure to appreciable concentrations of benzene and acute myelogenous leukemia, exposure of refinery workers has been studied for many years. To date, no extensive industrial hygiene exposure analyses for historical benzene exposure have been performed, and none have focused on the airborne concentrations in the workplace at specific refineries or for specific tasks. In this study, the authors evaluated the airborne concentrations of benzene and their variability over time at the ExxonMobil refinery in Baton Rouge between 1977 and 2005. Refinery workers were categorized into 117 worker groups using company job descriptions. These 117 groups were further collapsed into 25 job categories based on similarity of measured exposure results. Results of 5289 personal air samples are included in this analysis; 3403 were considered nontask (>or= 180 min) personal samples, and 830 were considered task-related (< 180 min) personal samples; the remainder did not fit in either category. In general, nontask personal air samples indicated that exposures of the past 30 years were generally below the occupational exposure limit of 1 ppm, but there was only a small, decreasing temporal trend in the concentrations. The job sampled most frequently during routine operations was process technician and, as broken down by area, resulted in the following mean benzene concentrations: analyzers (mean = 0.12 ppm), coker (mean = 0.013 ppm), hydrofiner (mean = 0.0054 ppm), lube blending and storage (mean = 0.010 ppm), waste treatment (mean = 0.092 ppm), and all other areas (mean = 0.055 ppm). Task-based samples indicated that the highest exposures resulted from the sampling tasks, specifically from those performed on process materials; in general, though, even these tasks had concentrations well below the STEL of 5 ppm. The most frequently sampled task was gauging (mean = 0.12 ppm). Task-related exposures were also similar across job categories for a given task, with a few exceptions. This study thus provides a task-focused analysis for occupational exposure to benzene during refinery operations, which can be insightful for understanding exposures at this refinery and perhaps others operated since about 1975. PMID- 19544136 TI - Comparison of daily and retrospective reports of vaginal sex in heterosexual men and women. AB - This study examined the agreement between daily and retrospective reports of vaginal sex over a two-month period in a sample of 376 heterosexually active men and women. We also examined whether gender, age, or method of daily data collection (self-administered vs. interviewer administered) were related to agreement between daily and retrospective reports. Both counts and categorical measures of frequency of the behaviors were examined. There were no gender, age, or data collection method effects. When measured as a count, participants reported more instances of vaginal intercourse in the retrospective reports than on the daily reports. In contrast, comparison of retrospective categorical measures of frequency to daily reports showed considerable variability. Possible reasons for the over-reporting of counts of vaginal sex in retrospective reports are explored. PMID- 19544138 TI - Paraneoplastic cerebellar degeneration: initial presentation in a patient with anaplastic T-cell lymphoma, associated with ichthyosiform cutaneous lesions. PMID- 19544139 TI - Changes in serum tryptophan catabolism as an indicator of disease activity in adult T-cell leukemia/lymphoma. PMID- 19544140 TI - Report on the First International Workshop on Interim-PET-Scan in Lymphoma. AB - An international workshop, under the auspices of Group d'Etude des Lymphomes de l'Adulte (GELA) took place in Deauville, France, in April 3-4, 2009. The European experts with a published or personal experience on interim-PET in lymphoma were invited to the meeting. The aim of the workshop was twofold: (1) to reach a consensus on simple, reproducible criteria for interim-PET interpretation in Hodgkin lymphoma and diffuse large B-cell lymphoma (DLBCL), and (2) to launch two or more international validation studies, in an attempt to validate these rules. We concisely report here the minutes of the meeting and the conclusions/statements that have been reached. PMID- 19544142 TI - Covert face priming reveals a 'true face effect' in a case of congenital prosopagnosia. AB - Previous research indicates that individuals with congenital prosopagnosia (CP) fail to demonstrate significant priming from faces to related names in covert recognition tasks. The interpretation has been that CP precludes the ability to acquire face representations. In the current study we replicated this important finding. In addition, we also demonstrated significant 'true face effect' in a CP patient, where face primes that matched the probe names facilitated reaction times compared to unrelated face primes. These data suggest that some individuals with CP may possess degraded face representations that facilitate the priming of a person's identity, but not semantic associates. PMID- 19544141 TI - Sustained response of primary cutaneous CD30 positive anaplastic large cell lymphoma to bexarotene and photopheresis. PMID- 19544143 TI - Naming performance in two bilinguals with frontal vs. temporal glioma. AB - Two bilingual patients had World Health Organization Grade II Gliomas removed from a language area, one in the left mesiofronto-cingular region and one in the left postero-temporal region. They performed a picture naming task in their two languages before their surgery and afterwards. Both patients showed slowness in naming in their first language but different patterns of naming performance across their first and second language. Their patterns depended upon the site of their lesion and their language experience. These data, from brain-damaged, bilingual adult patients, contribute to the neuropsychological literature on brain organization and plasticity, and highlight the importance of assessing naming speed to obtain a better understanding of impairment and recovery mechanisms. PMID- 19544144 TI - Seizures in corticobasal degeneration: a case report. AB - Seizures are relatively common in Alzheimer disease (AD) and other neurodegenerative disorders. To our knowledge, however, there have been no reports of seizures associated with corticobasal degeneration (CBD). We describe a patient with brain biopsy features suggestive of CBD whose course was complicated by complex partial seizures with secondary generalization. Thus, the occurrence of seizures in a patient with dementia should not exclude the diagnosis of CBD. PMID- 19544145 TI - Longitudinal relations among negative affect, substance use, and peer deviance during the transition from middle to late adolescence. AB - The transition from middle to late adolescence brings challenges that increase risk for emotional, behavioral, and social problems. The nature of the associations among these types of problems is poorly understood. This National Institute on Drug Abuse-funded study examined longitudinal relations among negative affect, substance use, and peer deviance from ages 16 to 18 years. Multiwave youth and parent questionnaire data collected from 429 sixth graders (222 girls) and their families residing in the rural Midwestern United States and recruited in 1993 were analyzed via structural equation modeling. Consistent with the self-medication hypothesis, negative affect statistically predicted increased substance use over time. Implications for theory and prevention are discussed and the study's limitations are noted. PMID- 19544146 TI - Assessing the reliability of scores produced by the substance abuse subtle screening inventory. AB - The Substance Abuse Subtle Screening Inventory (SASSI) is a 10 scale indirect screening instrument used to detect substance use disorders. The current meta analytic study described reliability reporting practices across 48 studies involving the SASSI. Reliability generalization methods were then employed to evaluate typical score reliability for the screening measure. Results showed approximately 73% of studies did not report reliability estimates. Analysis of data from the remaining studies revealed adequate reliability for the total scale (alpha = .87) and face valid scales (FVA alpha = .88 and FVOD alpha = .92), but substantially lower reliability estimates for the indirect scales (range of alpha = .23-.65). The study's findings underscore the need for improved reliability reporting for the SASSI and suggest cautious use of the measure, especially its indirect scales, as an indicator of problematic substance use/abuse in clinical settings. PMID- 19544147 TI - Alcohol screening scores predict risk of subsequent fractures. AB - The Alcohol Use Disorders Identification Test-Consumption (AUDIT-C; 0-12 points) was included on health surveys in a cohort of 32,622 general medicine outpatients from seven US Department of Veterans Affairs (VA) hospitals. Cox proportional hazards models were used to estimate the risk of fracture (mean follow-up = 1.6 years) by AUDIT-C category. After adjusting for confounders, AUDIT-C scores of 8 9 and 10-12 were associated with significantly increased risks for subsequent fractures, HR (95% CI) = 1.37 (1.03 to 1.83) and 1.79 (1.38 to 2.33) respectively. These results can be used to provide feedback to patients linking their alcohol screening scores to medical outcomes-a critical component of evidence-based brief counseling for alcohol misuse. The study's limitations are noted. PMID- 19544149 TI - Killing time with enjoyment: a qualitative study of initiation into injecting drug use in north-east India. AB - Manipur and Nagaland are north-east Indian states characterized by a high prevalence of injecting drug use and HIV in a context of socio-economic underdevelopment and political instability. This qualitative study aims to increase understanding of the contextual factors associated with initiation into injecting drug use in these two states. Forty semi-structured in-depth interviews were conducted among injecting drug users (10 women, 30 men) aged 18-28 years in mid-2006. The interview transcripts were thematically analyzed. All participants were initiated into injecting by another person, most commonly a friend and often in the context of well-established social networks. Most were poly-drug users and unsafe injecting practices were frequently associated with the initiation experience. The subjective reasons for deciding to inject were pleasure-seeking, influence of peers, and economic reasons. We hypothesize that initiation into injecting in this part of the world is also linked to ideas of masculinity, and that young men engage in drug use in order to fill a social vacuum created by limited opportunities to meaningfully engage in adult roles within the community. The findings from this study suggest that harm reduction programs need to target (noninjecting) drug users, and that existing social networks could be creatively used to extend the reach of these programs. PMID- 19544148 TI - Ethnic differences in substance use, sexual risk behaviors, and psychosocial factors in a sample of heterosexual methamphetamine users. AB - Ethnic differences in measures of substance use, sexual risk behaviors, and psychosocial factors (depression, stigma, self-esteem) were examined in a sample of 402 heterosexual methamphetamine users (55.0% Caucasian, 29.9% African American, 15.1% Latino) who participated in a sexual risk reduction intervention between June 2001 and March 2005 in San Diego, California. Participants were primarily male (67%) and noncollege graduates (72%) and belonged to the low income group (66%). African Americans were older when they first used methamphetamine and had used fewer grams in the past 30 days; Caucasians were more likely to inject. A larger percentage of African Americans reported anonymous sex partners in the past 2 months. African Americans reported lower levels of social stigma, and Caucasians reported lower self-esteem. Limitations and potential applications of these findings to prevention and treatment programs for ethnic minority populations are discussed. PMID- 19544150 TI - Typologies of alcohol use in White and African American adolescent girls. AB - This study examined typologies of alcohol use among 2,948 White and African American adolescent girls using data from the National Longitudinal Study of Adolescent Health. Self-report data were collected on frequency and quantity of alcohol use, negative consequences, and high-risk drinking behaviors, as well as co-occurring internalizing and externalizing problems. Latent class analysis revealed a four-group typology for White girls and a three-group typology for African American girls. Problematic drinkers reported having more internalizing and externalizing problems in both racial groups. The study's limitations and implications are discussed. PMID- 19544151 TI - Markedly elevated serum alkaline phosphatase level in an uncomplicated pregnancy. AB - In normal pregnancy, although maternal serum alkaline phosphatase increases and reaches the two-fold of non-pregnant serum level, above this level, hepatic, renal, osteal diseases and malignancies should be kept in mind. We report a case of 28-year-old pregnant woman with increased maternal serum level of alkaline phosphatase activity (17-fold) due to an increased level of placental isozyme. She delivered vaginally a healthy female baby. After delivery, maternal serum alkaline phosphatase activity returned to normal reference range. The aim of this case report is to show this exaggerated isozyme activity of alkaline phosphatase in pregnancy, and to differentiate it from other causes of elevated level of alkaline phosphatase. PMID- 19544153 TI - The effect of caesarean section on self-esteem amongst primiparous women in South Western Nigeria: a case-control study. AB - OBJECTIVE: This study aims to assess the level of self-esteem of newly delivered mothers who had caesarean section (CS) and evaluate the sociodemographic and obstetrics correlates of low self-esteem in them. METHODS: Newly delivered mothers who had CS (n = 109) and who had spontaneous vaginal delivery (SVD) (n = 97) completed questionnaires on sociodemographic and obstetrics variables within 1 week of delivery. They also completed the Rosenberg self-esteem scale. RESULTS. Women with CS had statistically significant lower scores on the self-esteem scale than women with SVD (p = 0.006). Thirty (27.5%) of the CS group were classified as having low self-esteem compared with 11 (11.3%) of the SVD group (p = 004). The correlates of low self-esteem in the CS group included polygamy (odd ratio (OR) 4.99, 95% confidence interval (95% CI) 1.62-15.33) and emergency CS (OR 4.66, 95% CI 1.55-16.75). CONCLUSIONS: CS in South-Western Nigerian women is associated with lowered self-esteem in the mothers. PMID- 19544152 TI - Effect of plasma proteins on buprenorphine transfer across dually perfused placental lobule. AB - OBJECTIVE: The aim of this investigation is to determine the effect of human serum albumin (HSA) and alpha-acid glycoprotein (AAG) on buprenorphine (BUP) transplacental transfer and distribution. METHODS: The technique of dual perfusion of placental lobule (DPPL) was utilised. BUP was co-perfused with the marker compound antipyrine (AP). In each experiment, the radiolabelled isotopes [(3)H]-BUP and [(14)C]-AP were added to enhance their detection limits. Human plasma proteins, HSA and AAG, were added to both the maternal and fetal circuits separately and in combination at their physiological concentrations in maternal and fetal circulations close to term. RESULTS: Transplacental transfer of BUP, in absence of plasma proteins, is a two-step process: the first is its uptake by the syncytiotrophoblast from the maternal circuit, and the second is its transfer/release from the tissue to the fetal circuit. The addition of HSA to the perfusion medium affected only the second step of BUP transfer, but AAG affected both steps. The combined effect of HSA and AAG was not different from that observed in presence of the latter alone. CONCLUSIONS: Binding of BUP to circulating AAG could have an important role in the transfer of the drug from the maternal to fetal circulation. PMID- 19544157 TI - Amniotic fluid prostaglandin F2 increases even in sterile amniotic fluid and is an independent predictor of impending delivery in preterm premature rupture of membranes. AB - OBJECTIVE: To determine whether amniotic fluid (AF) concentration of prostaglandins (PGs) increases in patients with intra-amniotic inflammation and/or proven AF infection in preterm PROM, and can predict impending delivery. METHODS: AF PGF2a concentrations were determined by ELISA in 140 singleton pregnancies with preterm premature rupture of membranes (PROM) (< or =35 weeks). AF was cultured for aerobic and anaerobic bacteria, and genital mycoplasmas. Intra-amniotic inflammation was defined as an elevated AF matrix metalloproteinase-8 concentration (>23 ng/ml). Results. (1) Patients with intra amniotic inflammation and a negative AF culture had a significantly higher median AF PGF2a than those without intra-amniotic inflammation and with a negative culture (p < 0.001); (2) However, there was no difference in the median AF PGF2a between patients with intra-amniotic inflammation with a negative culture and those with culture-proven AF infection (p > 0.1); (3) Patients with an elevated AF PGF2a had a significantly shorter interval-to-delivery than those with a low AF PGF2a (< or =170 pg/mL) (p < 0.001); (4) An elevated AF PGF2a (< or =170 pg/mL) concentration was a significant predictor of the duration of pregnancy after adjusting for gestational age and AF inflammation/infection (p < 0.005). Conclusions. AF PGF2a (> or =170 pg/mL) concentration increased in patients with intra-amniotic inflammation regardless of AF culture results. Moreover, an elevated AF PGF2a concentration was an independent predictor of impending delivery in preterm PROM. PMID- 19544160 TI - Does eczema lead to asthma? AB - The nature of the relationship between eczema, asthma, and allergic rhinitis has been controversial. It has been commonly held that these disorders, while sharing genetic and environmental risk factors, are unrelated disorders that may develop sequentially along an atopic pathway. Conversely, the link between eczema and these later-onset respiratory disorders may be causal. This review examines the relationship between eczema, asthma, and allergic rhinitis in the context of the atopic march, the skin barrier, and recent developments in eczema genetics; and we propose that the relationship is causal. We describe a plausible biological pathway with eczema as the first step in a progressive atopic march that over time leads to asthma and/or allergic rhinitis. Such a pathway has implications for our understanding of these disorders and steps that might be made to prevent the development of asthma in particular. We propose that intervention studies in eczema should be conducted to confirm or refute this causal relationship. Such studies may materially improve the quality of life of eczema patients and will have important public health benefits if the interventions lead to a reduction in the burden of asthma. PMID- 19544161 TI - What affects asthma medicine use in children? Australian asthma educator perspectives. AB - The global burden of childhood asthma is significant. Health care systems are faced with increasing financial costs, while children with asthma and their caretakers are faced with poorer physical health, emotional health, and quality of life. Despite the availability of effective treatment, the quality use of asthma medicines in children remains suboptimal. An investigation was conducted to explore issues related to children's asthma medicine usage from the perspective of the health care professional. Although current literature has elicited the views of caretakers and children, the health care professional viewpoint has been relatively unexplored. Semi-structured qualitative interviews were conducted with a convenience sample of 21 Australian asthma educators. Interviews were audiotaped and transcribed, and transcripts were thematically analyzed with the assistance of NVivo 7. Emergent themes associated with health care professionals, parents, medicines, children, and educational resources were found. Major issues included a lack of information provided to parents, poor parental understanding of medicines, the high cost of medicines and devices, child self-image, the need for more child responsibility over asthma management, and the lack of standardization, access to, and funding for educational resources on childhood asthma. There are multitudes of key issues that may affect asthma medicines usage in children. This research will help inform the development of educational tools on the use of medicines in childhood asthma that can be evaluated for their effectiveness in getting key messages to target audiences such as children, caretakers, and teachers. PMID- 19544162 TI - Resolution of corticosteroid-induced diabetes in allergic bronchopulmonary aspergillosis with omalizumab therapy: a novel approach. PMID- 19544164 TI - The response to bronchodilators in adults is not predictive of bronchial hyperreactivity. AB - BACKGROUND: In some subjects with suspected asthma who have normal spirometry, administration of bronchodilators (BD) improves expiratory flow rates. The predictive value of this phenomenon in adults is not known. OBJECTIVES: To evaluate the predictive value of the response to BD for bronchial hyper responsiveness (BHR) using the metacholine challenge test (MCT). Patients and methods. The study population included 62 non-smoking adult patients (41.9% women) 29.5 +/- 15.5 years of age (range 18-64 years) with suspected asthma with normal spirometry that underwent MCT within 1 week. The response to BD (200 mu g inhaled salbutamol) was compared between subjects with positive and negative MCT using cutoff levels of provocative concentrations of metacholine causing a 20% decrease in forced expiratory volume in 1 second (FEV(1)) (PC(20)) of 4 and 8 mg/mL. RESULTS: Mean (+/- SD) baseline FEV(1) was 87.8 +/- 12% of predicted. After BD administration the mean FEV(1) increased by 4.3 +/- 3.9%. The prevalence of BHR was 17.7% and 25.8% for PC(20) for PC(20) of 4 mg/mL and 8 mg/mL, respectively. The post-BD FEV(1) increment for subjects with positive and negative MCT tests was 3.9% +/- 3.3% versus 4.4% +/- 4.1%, respectively; p = 0.89, using cutoff of 4 mg/mL. The corresponding figures for cutoff of 8 mg/ml were 4.3% +/- 3.1% vs. 4.3% +/- 4.2%, respectively; p = 0.8465. There was no correlation between post-BD FEV(1) increment and PC(20) values in patients with positive MCT test for the above-mentioned cutoff levels (correlation coefficient r = 0.1645, p = 0.6289; and r = 0.2417, p = 0.4051, respectively). CONCLUSIONS: In adults with suspected asthma who have normal spirometry, the response to BD cannot be used to predict BHR. PMID- 19544163 TI - Ethnic disparities in asthma morbidity in Chicago. AB - RATIONALE: The role of ethnicity and socioeconomic status in explaining variations in asthma morbidity is unclear. OBJECTIVES: To describe the magnitude of ethnic disparities in asthma morbidity in Chicago and to determine whether differences in socioeconomic status explain these disparities. METHODS: We conducted a survey of 561 school-age children and 353 young adults with asthma and measured their self-reported ethnicity, socioeconomic status (using 11 variables), and asthma morbidity (symptom frequency, asthma-specific quality of life, and frequency of severe asthma exacerbations). MEASUREMENTS AND MAIN RESULTS: White children and adults had better asthma-specific quality of life and fewer severe asthma exacerbations compared to black and Hispanic children and adults. White children also had fewer days with asthma symptoms, but among adults there were no ethnic differences in the frequency of asthma symptoms. Socioeconomic status explained a large portion of the ethnic disparities in asthma quality of life but explained little of the disparities in other aspects of asthma morbidity. CONCLUSIONS: There are large disparities across ethnic groups in Chicago in asthma quality of life and in the frequency of severe exacerbations. Differences in socioeconomic status do not fully explain these disparities. PMID- 19544165 TI - Identifying preschool children with asthma in Orange County. AB - Airway changes related to childhood asthma occur early in the disease process. This pilot study focuses on the validation of the Breathmobile Case Identification Survey (BCIS) in preschool-age children in Orange County, CA. Fifty-two children from low-income Spanish-speaking families participated in the study. Thirteen children were identified as possibly having asthma from the survey results compared with 20 children diagnosed by an asthma specialist. We found that the complete seven-question survey had a sensitivity of 0.65 and a specificity of 0.94. An abbreviated three-question version had a sensitivity of 0.70 and a specificity of 0.84. Our data suggest that the abbreviated BCIS, which is simple and easily analyzed, may be a useful tool in identifying young children who are at risk for asthma and need further evaluation and appropriate therapy. PMID- 19544166 TI - A randomized study comparing the effect of loratadine added to montelukast with montelukast, loratadine, and beclomethasone monotherapies in patients with chronic asthma. AB - BACKGROUND: Loratadine added to montelukast has been suggested to improve endpoints of asthma. OBJECTIVE: This study investigated the additive effects of concomitant montelukast and loratadine when compared with montelukast, loratadine, and inhaled beclomethasone monotherapies in asthma. Methods. Patients (N = 406) were 15 to 65 years of age with a forced expiratory volume in 1 second (FEV(1))-predicted of 50% to 85%, FEV(1) reversibility > or = 15%, and a minimal level of daytime symptoms and beta -agonist use. This three-part 2X2 crossover study consisted of two double-blind 6-week treatment periods where patients were administered once daily oral montelukast 10 mg, loratadine 10 mg, montelukast 10 mg + loratadine 10 mg, or twice daily inhaled beclomethasone 200 mu g. A subsequent 48-week extension study compared montelukast + loratadine with beclomethasone. The primary endpoint was the percentage change from baseline in FEV(1). RESULTS: Over 6 weeks of double-blind treatment, significant improvements (p < 0.05) in the primary endpoint of FEV(1) were seen for montelukast + loratadine versus loratadine (least-square mean percentage-point difference of 5.8%), beclomethasone versus montelukast + loratadine (2.35%), montelukast versus loratadine (5.94%), and beclomethasone versus montelukast (4.65%); a numerical improvement (p = 0.054) was seen for montelukast + loratadine versus montelukast (1.60%). Significant improvements for montelukast + loratadine versus montelukast were seen in some secondary endpoints (evening peak expiratory flow, nocturnal asthma symptom score, nocturnal awakenings, and asthma-specific quality of life) but not others. Significant improvements in most endpoints except daytime asthma symptoms score were seen for montelukast + loratadine versus loratadine. In the extension study, both montelukast + loratadine and beclomethasone improved several endpoints. All treatments were generally comparable in the percentage of patients with clinical and laboratory adverse experiences. CONCLUSION: In this study, the addition of loratadine to montelukast produced a small numerical, but not statistically significant, improvement in FEV(1) and, in general, no consistent improvement in other asthma endpoints. No improvement of montelukast + loratadine versus beclomethasone was seen in any endpoint. PMID- 19544167 TI - Smoking affects eotaxin levels in asthma patients. AB - BACKGROUND: Chronic airway inflammation is most important pathological finding in asthma. Cigarette smoking may modify type of inflammation as well as may influence disease severity and response to the treatment. OBJECTIVE: Thus the aim of this study was to investigate whether cigarette smoking may have an influence on the levels of eotaxin-1, eotaxin-2, eotaxin-3 and IL-5 in patients with stable mild/moderate asthma. METHODS: 45 steroid naive asthmatics (mean age: 55.2 +/- 2.2 yrs) and 23 "healthy" smokers and non-smokers control subjects (mean age: 54.4 +/- 9.7 yrs) were investigated. Asthmatics were divided into two subgroups according to their smoking histories: asthmatic smokers (n = 19) who currently smoke and have a history of > 10 pack-years and asthmatic never-smokers (n = 26). BAL and induced sputum were performed. Cytospins of induced sputum and BAL were stained with May-Grunwald-Giemsa for differential cell counts. Eotaxin-1, eotaxin 2, eotaxin-3 and IL-5 concentrations in serum, sputum and BAL supernatant was measured using a commercial ELISA kit. RESULTS: In sputum supernatant from asthma smokers was significantly higher concentration of eotaxin-1 than in non-smokers asthmatics (203.4 +/- 10.0 vs. 140.2 +/- 9.5 respectively, p < 0.05). In non smokers asthma patients levels of BAL eotaxin-1 strongly related to percent and absolute numbers of BAL eosinophils and neutrophils (Rs = 0.737 and Rs = 0.514 respectively, p < 0.05). The number and percent of sputum neutrophils and eosinophils, obtained from smokers asthmatics, significantly correlated with eotaxin-2 concentration in sputum supernatant (Rs = 0.58 and Rs = 0.75 respectively, p < 0.05). IL-5 levels in the serum and sputum from asthmatic never smokers were significantly higher than they were from asthmatic smokers and "healthy" smokers. Asthmatic never-smokers showed a significantly higher amount of IL-5 in serum and sputum than the asthmatic smokers showed. CONCLUSIONS: This study showed the elevated levels of sputum eotaxin-1 as well as serum, sputum and BAL eotaxin-2 in asthmatic smokers without a significant increase of eosinophils compared to asthmatic never-smokers. The eotaxin concentrations were related not only with number of eosinophils but also with the number of neutrophils in all the studied tissue compartments. The data herein permits a suggestion that smoking may influence change in asthmatic airway inflammation by stimulating the production of eotaxins. PMID- 19544168 TI - Does influenza vaccination improve pediatric asthma outcomes? AB - OBJECTIVE: Controversy exists regarding the effectiveness of influenza vaccination in preventing influenza-related asthma exacerbations in the pediatric population. While yearly influenza immunization is widely recommended for children with asthma, there is currently little evidence to support this practice. Several studies have demonstrated no measurable benefit in asthma outcomes. This study sought to determine whether influenza vaccination status is associated with indicators of asthma morbidity within the military pediatric population. METHODS: A survey was conducted of patients 3 to 18 years of age with a diagnosis of asthma enrolled in the pediatric clinics of Brooke Army Medical Center, Fort Sam Houston, Texas, and Wilford Hall Medical Center, Lackland Air Force Base, Texas. Management practices and outcomes for 80 children were evaluated. Data were analyzed using the statistical package SPSS version 12 (SPSS Inc., Chicago). Univariate analyses were performed to identify associations between influenza vaccination, selected demographic variables and asthma exacerbation defined by oral steroid prescription, hospital visits, and unscheduled clinic or emergency department visits for asthma symptoms. Logistic regression analyses were conducted to detect possible confounding variables. RESULTS: In the univariate analyses, current influenza vaccination status was associated with a significant reduction of oral steroid use in the 12 months before the survey. This relationship was appreciated to a lesser extent with emergency department or unscheduled clinic visits in the last 12 months. No significant differences were found regarding the distribution of influenza vaccination status across selected demographic variables. In the multivariate analyses, current influenza vaccination status was independently associated with significantly decreased odds of using oral steroids in the previous 12 months. There was no evidence of confounding or effect modification. CONCLUSIONS: This study suggests influenza vaccination is associated with fewer asthma exacerbations. After controlling for several potential confounding variables, administration of influenza vaccine was associated with a protective effect against indicators of asthma exacerbations. Our results indicate that children with asthma in the military beneficiary population may benefit from annual influenza vaccination. PMID- 19544169 TI - Allergic and nonallergic rhinitis: can we find the differences/similarities between the two pictures? AB - The diagnostic challenge of rhinitis is to determine the etiology, specifically whether it is allergic or nonallergic. We therefore evaluated the general features of patients with allergic (AR) and nonallergic rhinitis (NAR), as well as health-related quality of life (HRQoL). The study group consisted of 323 patients (201 F/122 M) with a mean age of 31.79 +/- 12.64 years. Almost two thirds of the population had AR (63.5%). Neither the demographic characteristics nor the duration of rhinitis was different between the two groups. Total immunoglobulin E was significantly higher in AR. Although both groups displayed a mild-intermittent rhinitis profile, patients with AR had more seasonal and NAR had more perennial symptoms (p = 0.01). Frequency of nasal obstruction was comparable in both groups, whereas patients with AR significantly complained of rhinorrhea (86.8%), followed by nasal obstruction, sneezing, and nasal itching compared with the NAR group. Conjunctivitis and sinusitis were more prominent in the AR than NAR group (p = 0.01). However, the prevalences of asthma and bronchial hyperreactivity were not different, as well as the other allergic or systemic comorbidities. Furthermore, the impairment in HRQoL was similar in both groups, using a generic questionnaire- Short form-36 (SF-36). In conclusion, although the allergy test results still remain the only relevant difference, the diagnosis of NAR is important as it has many differences/similarities with AR and is seen almost half as often as AR in patients with chronic rhinitis. PMID- 19544170 TI - Airway hyperresponsiveness: a comparative study of methacholine and exercise challenges in seasonal allergic rhinitis with or without asthma. AB - BACKGROUND: Asymptomatic airway hyperreactivity in allergic rhinitis is a risk factor for later development of asthma. Although non-specific bronchial hyperresponsiveness (BHR) has been measured by several stimuli, the most appropriate measurement technique still remains unclear. OBJECTIVE: To investigate whether an exercise challenge can be used to predict BHR in seasonal allergic rhinitis patients with or without asthma and to compare this bronchial reactivity with a methacholine challenge technique. METHODS: Forty-six consecutive patients with seasonal allergic rhinitis only (n = 31) and with both seasonal allergic rhinitis and asthma (n = 15) were included in the study during the pollination period. Subjects underwent first methacholine (mch) and then exercise challenge testing (ECT). There was a 1-week interval between the tests. ECT was performed on a bicycle ergometer. Positive result was defined as a 15% decrease in forced expiratory volume in 1 second (FEV(1)) post-exercise. A patient's bronchial reactivity to methacholine was considered as hyperresponsive if PC(20) was less than 8 mg/mL. RESULTS: Mch PC(20) values were significantly lower in patients with both rhinitis and asthma (p < 0.062). Among the 46 patients, mch PC(20) values were significantly different between patients who had positive and negative exercise challenge tests (p = 0.007). All patients with rhinitis alone had a negative ECT and 10 had a positive mch challenge. Change in FEV(1) values after ECT was significantly higher in patients with both rhinitis and asthma compared to those with rhinitis alone (p = 0.009). There was a significant relation between positivity of mch and exercise challenges (p = 0.025). ECT positivity was found to be a significant confounding factor in the diagnosis of asthma (p = 0.001). Specificity and sensitivity values were 100% and 24% for ECT and 68% and 100% for mch, respectively. CONCLUSION: Exercise challenge presents poor diagnostic value for detecting bronchial responsiveness in individuals with allergic rhinitis alone during the pollen season. PMID- 19544172 TI - Detection of undiagnosed and poorly controlled asthma in a hospital-based outpatient pediatric primary care clinic using a health risk assessment system. AB - OBJECTIVE: To determine the rate of undiagnosed and poorly controlled asthma detected by a computerized health risk assessment (HRA) survey system in an urban pediatric hospital-based outpatient teaching clinic. METHODS: A software-based HRA system uses survey answer patterns to identify children with (1) probable, (2) uncontrolled, and (3) moderate to severe asthma. Parents of patients > or = 2 years of age were asked by clinic staff to complete the touch screen computer survey before seeing their physician from August 2005 through July 2006. RESULTS: The HRA survey predicted 26% (282/1,098) to have probable asthma. Of these, 51% (144/282) were controlled and the parents reported a previous diagnosis of asthma; 14% (40/282) were controlled and the parents did not report a previous diagnosis of asthma; 25% (71/282) were uncontrolled and the parents reported a previous diagnosis of asthma; and 10% (27/282) were uncontrolled and no previous diagnosis of asthma was reported by the parents. Among active cases completing the baseline version survey (N = 217), 68% reported emergency department (ED) visits / hospitalizations in the last 2 years (44% > or =2), while 59% reported missed school days during the previous year (23% > or =5 days). Impairment, as defined by the 2007 National Heart, Lung, and Blood Institute/National Asthma Education and Prevention Program (NHLBI/NAEPP) asthma guidelines, tended to be higher in patients without a previous diagnosis of asthma, per parental report, but this trend only achieved significance in two measures: daytime symptoms > or =2 days per week in the last 4 weeks (p = 0.028) and more than 5 missed school days in the past year (p = 0.001). CONCLUSION: A previously validated HRA system can consistently identify a high rate of undiagnosed and poorly controlled asthma in an urban pediatric hospital-based teaching clinic. The utility of such a system would be to reduce missed opportunities for delivery of care and morbidity for the patients who currently have undiagnosed and/or uncontrolled asthma in the pediatric primary care outpatient setting. PMID- 19544171 TI - Enrolling adolescents in asthma research: adolescent, parent, and physician influence in the decision-making process. AB - BACKGROUND: The factors influencing family decisions to participate in adolescent asthma research are not well understood. Legal and ethical imperatives require adolescent research participation to be voluntary. While parents and adolescents often agree about research decisions, disagreements may also occur with relative frequency. Physician recommendations are also known to influence research participation decisions. Little attention has been given to how these dynamics may affect adolescents' involvement in decisions to participate in research. OBJECTIVE: To examine the influence of family and physician-investigator relationships and recommendations on adolescent asthma clinical research participation decisions. METHODS: A statewide community sample of 111 adolescents 11 to 17 years of age, with a diagnosis of asthma, and their parents participated in this study. Adolescents received a medical evaluation from an asthma specialist and then the family was offered participation in a hypothetical asthma clinical trial. By random assignment, the research study was presented by either the same or an unknown asthma specialist, and half the families in each group also received affirmative recommendations from the asthma specialist to participate in the hypothetical asthma clinical trial. Parents and adolescents made initial private decisions about participating in the trial. Then, following a family discussion of the clinical trial, a final research participation decision was made. RESULTS: Thirty-three percent of parents and adolescents initially disagreed about the research participation decision. When disagreements occurred, final decisions followed the parents' initial views except when the physician-investigator was known and a recommendation was made. Families with initial disagreement about participating were less likely to enroll when the investigator was unknown or when no recommendation was made. Adolescents who initially disagreed with parents' views were less likely to concur with the final research participation decision, felt less comfortable, and were less likely to feel they influenced the decision. CONCLUSIONS: Parents' views on research decisions take precedence over adolescents' views in most circumstances. Physician-investigator relationships may reduce parental resistance to participation and enhance adolescent decision-making autonomy when research participation is desired by the adolescent. PMID- 19544173 TI - The impact of normalization of esophageal acid profile by incremental protein pump inhibitors dosing in difficult asthma patients with proven gastro-esophageal acid reflux. AB - BACKGROUND: GER is common in patients with asthma and some, although not all, studies have shown benefit from identification and treatment of GER. METHODS: patients with persistent symptoms after optimisation of asthma therapy underwent pH monitoring and adjustment of GER therapy based on the results of repeated pH monitoring. Gastrointestinal symptom scores and asthma therapy requirements were recorded. RESULTS: Over a 2 year period, 51 patients with a definite diagnosis of asthma underwent pH monitoring with GER being identified in 32 (63%). Normal oesophageal acid exposure was achieved in 11 patients, 7 requiring a daily PPI dose greater than 20 mg; 8 patients had persisting abnormal oesophageal acid exposure (OAE) despite daily PPI doses up to 80 mg. 13 patients declined further pH studies. 8 (73%) patients with normalisation of OAE had meaningful reductions in long term asthma therapy with 2 patients stopping and 2 patients reducing long term oral corticosteroids and 4 others halving the dose of inhaled corticosteroids. No patient who had persisting GER had asthma therapy reduced, neither did any of the group of patients in whom GER was not identified. CONCLUSION: Tailoring GER therapy with repeated pH studies had a major impact in a subgroup of patients, greater than any other intervention employed in our clinic over the same period. This uncontrolled data adds to the evidence that effective management of GER reduces asthma symptomatology and allows therapy to be reduced in a subgroup of patients with difficult to control asthma. PMID- 19544174 TI - Exploring the supportiveness and preparedness of child care settings for children with asthma: a needs assessment. AB - BACKGROUND: The role of child care centers has increased to extreme importance as the majority of young children are in some form of child care setting. Child care centers are increasingly faced with providing health-related care to children with asthma. PURPOSE: The purpose of this study was to examine child care workers' perceived knowledge and confidence and their perceptions of their child care setting's preparedness to handle asthma-related issues. Methods. The study used a cross-sectional postal survey design that followed a modified Dillman's protocol. Questionnaires were sent to staff in 235 randomly selected child care centers across four Ontario public health regions. The questionnaire assessed several asthma-related areas: identifying and caring for children with asthma, emergency plans for asthma, and confidence in ability to handle asthma-related issues. RESULTS: A total of 489 (69% overall response rate) questionnaires were returned and 180 centers (76.6%) of the 235 child care centers participated. Most child care workers (67%) cared for a child with asthma, but only 21% ever received formal training on caring for a child with asthma. When asked about having an emergency plan for a child having an asthma flare-up/attack, 52.3% of the workers reported that their center lacked such a plan or they were unaware of it. Many (45%) reported feeling uncertain about how to manage worsening asthma. The area of trigger identification and management was also an issue regarding child center preparedness: 43.6% of centers had a plan or process and 48% of staff felt uncertain of their abilities in this area. Child care workers who reported receiving formalized training were more likely to have higher confidence scores in their ability to handle asthma-related issues compared with those who did not receive training in many areas. CONCLUSIONS: Gaps in asthma care preparedness exist within the child care system. The provision of formalized learning opportunities is one strategy that could narrow these gaps. PMID- 19544175 TI - Outcomes after periodic use of inhaled corticosteroids in children. AB - BACKGROUND: Many children with persistent asthma use inhaled corticosteroids on a periodic basis. Clinical trials in adults suggest that periodic use of inhaled corticosteroids may be effective for patients with mild persistent asthma. However, scant information exists on the clinical outcomes of children with asthma who are using inhaled corticosteroids on a periodic basis in real-world settings. OBJECTIVE: This prospective cohort study compared clinical outcomes during a 12-month follow-up period between children with persistent asthma whose parents believed that they were supposed to use inhaled steroids either (a) periodically or (b) daily year-round at the start of the period. The clinical outcomes studied were (1) asthma-related emergency department (ED) visits or hospitalizations, (2) uncontrolled asthma based on health care and medication use, and (3) outpatient visits for asthma. PATIENTS AND METHODS: The study population included children with persistent asthma from two health plans whose parents reported that they were using inhaled corticosteroids during a baseline telephone interview. The interviews collected information on whether the children's parents believed they were supposed to use inhaled corticosteroids on a periodic or daily basis, as well as baseline asthma symptom status, sociodemographic, and behavioral variables. We used computerized databases to identify clinical events for each child during the 12 months after their baseline interview. Uncontrolled asthma was defined as any asthma-related ED visit or hospitalization, two or more oral steroid prescription fills, or four or more beta-agonists canisters filled during the 12-month period. We compared these outcomes between the periodic versus daily users of inhaled corticosteroids using logistic regression analyses. We conducted both (1) a traditional logistic regression analysis in which we adjusted for selection bias by including covariates such as age, asthma physical status, sociodemographic and behavioral variables, and history of asthma-related health care use during the year before interview and (2) an analysis using propensity scores to more fully adjust for selection bias. RESULTS: Of a total of 476 children in the study, 55% of parents believed their children were supposed to be using inhaled corticosteroids on a periodic basis and 45% believed their children were supposed to be using them daily year-round based on the baseline parent interview. At baseline, periodic inhaled corticosteroid users had less severe asthma than daily users based on several measures including better asthma physical status scores on the Children's Health Survey for Asthma (mean 87 +/- 16.0 vs. 81 +/- 17.4, p = < 0.0001). During the year before the baseline interview, periodic users compared with daily users were less likely to have an ED visit or hospitalization (10% vs. 23%, p = 0.0001) and less likely to have had five or more albuterol prescription fills (13% vs. 31%, p < 0.0001). During the follow-up year, those who believed inhaled steroids were for periodic use were less likely than those who believed inhaled steroids were for daily use to have an ED visit or hospitalization for asthma (OR 0.36, 95% CI: 0.18-0.73), even after adjusting for baseline asthma status and other covariates. Similarly, those who believed inhaled steroids were for periodic use were less likely to have uncontrolled asthma, OR 0.38 (95% CI: 0.24-0.62). Analyses using propensity score adjustment yielded similar results to the logistic regression analyses. CONCLUSION: Children whose parents believed they were supposed to use inhaled corticosteroids on a periodic basis had less severe asthma at baseline than those whose parents believed they were supposed to be using them daily. Periodic users were less likely than daily users to have adverse asthma outcomes during 1-year follow-up. This suggests that clinicians may be applying appropriate selection criteria by choosing patients with less severe asthma for periodic inhaled corticosteroid regimens. PMID- 19544176 TI - Genetic variation of myeloperoxidase gene contributes to atopic asthma susceptibility: a preliminary association study in Russian population. AB - BACKGROUND: Recently, we have shown that both antioxidant and oxidant genes are proper candidates for asthma susceptibility genes. OBJECTIVES: In the present study we investigated whether a common polymorphism -463G > A in the promoter of myeloperoxidase (MPO) gene, an enzyme producing hypohalogenic oxidants, is associated with the risk of bronchial asthma. METHODS: We studied 429 unrelated Russian subjects including 215 asthmatic patients and 214 sex- and age-matched healthy controls from Central Russia. The genotyping of the polymorphism -463G > A in the MPO gene was performed by the polymerase chain reaction and the restriction fragment length polymorphism assays. RESULTS: It was found that a carriage of a -463A allele is associated with decreased risk of asthma (OR 0.64 95%CI 0.44-0.91, p = 0.013). Furthermore, variant genotypes (-463GA + AA) of the MPO gene were associated with decreased risk of asthma (OR adjusted by age, gender, and immunoglobulin E (IgE) level was 0.63 95%CI 0.42-0.95), but at a borderline statistical significance (Bonferroni corrected p = 0.017). Further analysis revealed that both a -463A allele and the -463GA/AA genotypes are significantly associated with decreased risk of atopic asthma (p = 0.01). No association of the -463G > A polymorphism of the MPO gene with non-atopic asthma has been revealed. We also found that the allele -463A (OR = 0.47 95%CI 0.27 0.81, p = 0.01) and the -463GA + AA genotypes (OR 0.43 95%CI 0.24-0.78, p = 0.005) are significantly associated with decreased risk of late-onset atopic asthma (the disease onset after 30 years). No association of both allele and genotypes of the polymorphism -463G > A of the MPO gene with early-onset of atopic and non-atopic asthma (the disease before 30 years) was seen. CONCLUSIONS: The results of this study provide novel insights into pathogenesis of bronchial asthma. We put forward a suggestion about a possible mechanism by which the -463G > A polymorphism of the MPO gene is involved into pathogenesis of asthma. PMID- 19544177 TI - A comprehensive review of the literature on exposure to metalworking fluids. AB - An extensive literature review was conducted of studies with exposure measurements to metalworking fluids (MWFs). A database of 155 arithmetic means based on 9379 aerosol measurements from published studies was compiled. Weighted arithmetic means (WAMs) and their variance calculated across studies were summarized based on decade (prior to 1970s through 2000s), industry (auto, auto parts, small job shops, and others), operation (grinding and machining), and fluid type (straight, soluble, synthetic, and semisynthetic). Total mass and total extractable mass measurements that were simultaneously collected were compared. Average concentrations by size fractions and mass median aerodynamic diameters (MMADs) were also analyzed. Analysis of the WAMs indicated a reduction in exposure levels over time regardless of industry or type of operation or fluid, with mean levels prior to the 1970s of 5.4 mg/m(3), which dropped to 2.5 mg/m(3) in the 1970s, to 1.2 mg/m(3) in the 1980s, and to 0.5 mg/m(3) in the 1990s. No further reduction was seen in the 2000s. A comparison by industry, operation, and fluid type found no consistent patterns in the measurement results. The percent extractable mass in the total aerosol samples varied by fluid type, with an average 84% in straight fluids, 58% in synthetic fluids, 56% in soluble fluids, and 42% in the semisynthetic fluids. Exposure means from the thoracic fraction (0.3-0.5 mg/m(3)) were slightly less than those for total aerosol for both the 1990s and 2000s, the only decades for which thoracic data were available. Respirable means did not change from the 1980s to the 2000s (generally about 0.2-0.3 mg/m(3)). The MMADs of the MWF aerosols averaged 4-6 microm. These measurement data indicate a clear reduction of exposure levels over time. They will be used for the retrospective assessment of exposure levels to MWFs in a population-based, case-control study of bladder cancer. PMID- 19544178 TI - Do coping and social support predict depression and anxiety in patients undergoing hematopoietic stem cell transplantation? AB - This study examined whether different types of coping and social support predict anxiety and depression in 212 hematopoietic stem cell transplant (HSCT) recipients. Data were collected prior to and 6 months after HSCT. Coping, social support, and gender predicted 26% of the variance in pre-HSCT anxiety and 24% of the variance in pre-HSCT depression. Coping and social support did not explain significant post-HSCT anxiety or depression when controlling for pretransplant anxiety or depression. High use of acceptance/ resignation coping, cognitive avoidance coping, lower tangible support, and lower belonging support were related to increased pre-HSCT anxiety and depression. Approach coping was not related to pre-HSCT anxiety. Patients who use acceptance/resignation coping and report low levels of two types of social support prior to HSCT may require additional intervention before HSCT, as they are at higher risk for depression and anxiety. PMID- 19544179 TI - Religiousness and prostate cancer screening in African American men. AB - This study was designed to examine the relationship between religiousness (organized, nonorganized, and intrinsic) and religious problem solving (collaborative, deferring, and self-directing) in prostate cancer screening (PCS) attitudes and behavior. Men (N = 481) of African descent between the ages of 40 and 70 participated. Hierarchical regression analyses revealed that religiousness and self-directed problem solving were associated with PCS attitudes. Intrinsic religiousness was associated with PCS attitudes after controlling for health and organized religiousness. Religiousness was not associated with PCS behavior. Intrinsic religiousness may be an important dimension of religiousness to be considered in tailoring cancer interventions for individuals from faith-based communities. PMID- 19544180 TI - Sexual desire and satisfaction in sexual life affecting factors in breast cancer survivors after mastectomy. AB - The purpose of this study was to examine impact of treatment and personality characteristics, and support need for femininity and body image on sexual desire and sexual satisfaction of women with breast cancer. The sample consisted of 123 married women who previously underwent mastectomy and applied to medical oncology clinics of a university hospital. Sexual desire of the women in this study was low, and they were slightly satisfied with sexual life. Consequently, the women required supporting needs of femininity and body image. The result of this study could be utilized to determine sexual desire and satisfaction related to sexual life of women with breast cancer. PMID- 19544181 TI - Life after cancer in India: coping with side effects and cancer pain. AB - The article aims to understand the coping strategies of postsurgery head and neck cancer patients in Mumbai, India. A descriptive research design with a sample of 80 patients suffering from head and neck cancer was selected to analyze their coping strategies in relation to sociodemographic profile and illness characteristics. The findings of the study highlighted that the spiritual methods of coping (such as prayer and meditation, adopting a positive attitude) were the most frequently used mainstream coping strategy, apart from other traditional methods (such as taking medications, indulging in exercise and activities to divert one's attention, etc.) of coping. The findings of the study help to broaden the understanding of various psychosocial aspects faced by the patient in India and provide progressive recommendations to improve the quality of life of the patient suffering from cancer. PMID- 19544182 TI - Suddenly, a carriage appears: social support needs of Latina breast cancer survivors. AB - Through focus groups and individual interviews, data were gathered on the emotional, informational, and instrumental support needs of 22 immigrant Latina women. A thematic analysis revealed that participants who perceived to receive social support reported less psychological distress and better adjustment to breast cancer than those who did not perceive this support. Types and sources of support varied across survivorship stages. Many needs were related to financial, linguistic, and cultural barriers participants encountered in the course of the disease. Based on the findings, we conclude with several clinical recommendations to improve the quality of life in this medically underserved population. PMID- 19544183 TI - Treatment definition in complex rehabilitation interventions. AB - Rehabilitation research is challenged to improve its evidence base, which requires more precise and more consistent conceptualisation and measurement of treatment ingredients. This paper presents the steps in defining and specifying treatments towards the construction of a therapy manual for experimental interventions, or a coding system for observational treatment research. Specifying the active ingredients, identifying the behavioural operations that are aligned with each, and developing procedural details and materials are discussed and illustrated using two on-going trials for treatment of traumatic brain injury. The process of assessing treatment fidelity, that is, the extent to which interventions are delivered, received and used as intended, is also discussed. PMID- 19544184 TI - Carrying out research into outcomes. Foreword. PMID- 19544185 TI - Use of the Adult Attachment Interview to measure process and change in psychotherapy. AB - The Adult Attachment Interview (AAI), developed and extensively tested in the domain of developmental psychology, has relevance to psychotherapy research. The authors compare and contrast the ways that social psychologists and developmental psychologists have operationalized the concept of attachment security and discuss corresponding implications for psychotherapy research. In addition, they provide an overview of the AAI and its development, reliability, and validation in developmental psychology and summarize recent work linking AAI responses with distinctive types of psychopathology. A summary of recent work showing the AAI to be a useful indicator of positive outcome in psychotherapy for adults with diverse problems, including depression, borderline personality disorder, and posttraumatic stress disorder, is provided. PMID- 19544186 TI - Volunteer client attachment moderates the relationship between trainee therapist attachment and therapist interventions. AB - This study explored the association between attachment security of therapists (alone and in interaction with clients' attachment) and therapist interventions in early sessions of short-term psychotherapy. Trainee therapists and volunteer clients (N=24) in short-term therapy completed the Experiences in Close Relationship Scale (Brennan, Clark, & Shaver, 1998) as a measure of adult romantic attachment orientations. Therapist interventions were identified and related to client and therapist attachment orientation. Results indicated that in early therapy sessions client attachment moderated the relationship between therapist attachment and therapist interventions. Specifically, avoidantly attached therapists intervened with more directive interventions when clients were high in attachment avoidance. PMID- 19544187 TI - Using functional magnetic resonance imaging in psychotherapy research: a brief introduction to concepts, methods, and task selection. AB - Functional magnetic resonance imaging (fMRI) has become an increasingly important methodology in the study of psychotherapy outcome and process. In this article, the authors offer a brief introduction to the use of fMRI in psychotherapy research aimed primarily at the informed clinician or investigator and with the goal of facilitating an understanding of study design and interpretation of research findings. After introducing the method and offering a rationale for its use in the study of psychotherapy, the authors outline major issues in fMRI data collection and analysis and emphasize the central role of the tasks used during the imaging session as critical to the interpretation of findings. They discuss how task selection influences the conclusions that can be drawn from fMRI studies of psychotherapeutic intervention and close with recommendations and caveats for the consumer of fMRI/psychotherapy research. PMID- 19544188 TI - Fragment-based prediction of cytochromes P450 2D6 and 1A2 inhibition by recursive partitioning. AB - The evaluation of the ADME (absorption, distribution, metabolism, and excretion) properties of drug candidates is an important stage in drug discovery. To speed up the numerous tests carried out on large databases of compounds, the help of robust and accurate in silico filters is increasingly required. We propose here a method to build predictive and interpretable models for the prediction of cytochrome P450 (CYP) 1A2 and 2D6 inhibition using recursive partitioning (RP), a well-known technique for the construction of decision trees. The originality of the work is the use of several descriptions of the molecules in terms of fragments, i.e. the MACCS keys and five in-house fingerprints based on the electron density properties of fragments, employed to draw easily understandable structure-activity models. The classifiers reached performances of 87.5% and 76.5% of prediction on a validation set for CYP1A2 and CYP2D6, respectively. The analysis of the first nodes of the RP trees permits us to highlight some relations between the structural fragments and the inhibition of CYPs. PMID- 19544189 TI - Using chemical categories to fill data gaps in hazard assessment. AB - Hazard assessments of chemicals have been limited by the availability of test data and the time needed to evaluate the test data. While available data may be inadequate for the majority of industrial chemicals, the body of existing knowledge for most hazards is large enough to permit reliable estimates to be made for untested chemicals without additional animal testing. We provide a summary of the growing use by regulatory agencies of the chemical categories approach, which groups chemicals based on their similar toxicological behaviour and fills in the data gaps in animal test data such as genotoxicity and aquatic toxicity. Although the categories approach may be distinguished from the use of quantitative structure-activity relationships (QSARs) for specific hazard endpoints, robust chemical categories are founded on quantifying the chemical structure with parameters that control chemical behaviour in conventional hazard assessment. The dissemination of the QSAR Application Toolbox by the Organisation for Economic Cooperation and Development (OECD) is an effort to facilitate the use of the categories approach and reduce the need for additional animal testing. PMID- 19544190 TI - Molecular simulation of polycyclic aromatic hydrocarbon sorption to black carbon. AB - Strong sorption of hydrophobic organic contaminants to soot or black carbon (BC) is an important environmental process limiting the bioremediation potential of contaminated soils and sediments. Reliable methods to predict BC sorption coefficients for organic contaminants are therefore required. A computer simulation based on molecular mechanics using force field methods has been applied in this study to calculate BC sorption coefficients of polycyclic aromatic hydrocarbons (PAHs). The free energy difference between PAHs dissolved in water and in water containing a model structure of BC was calculated by thermodynamic integration of Monte Carlo simulated energies of transfer. The free energies were calculated with a hypothetical reference state that has equal free energies in both phases and is therefore cancelled in the calculated free energy difference. The calculated sorption coefficient of phenanthrene (log K(BC) = 5.17 +/- 0.54 L/kg C), fluoranthene (6.33 +/- 0.64 L/kg C) and benzo[a]pyrene (7.38 +/ 0.36 L/kg C) corresponded very well to experimental values available in the literature. Furthermore, an average spacing distance of 3.73 A between PAHs and BC was determined that is only slightly lower than an experimentally determined value of 4.1 A. The method applied in this study enables the calculation of the extent of PAH sorption to a soot surface for which no experimental values are available nor data for related compounds as required in quantitative structure activity relationships. PMID- 19544191 TI - How not to develop a quantitative structure-activity or structure-property relationship (QSAR/QSPR). AB - Although thousands of quantitative structure-activity and structure-property relationships (QSARs/QSPRs) have been published, as well as numerous papers on the correct procedures for QSAR/QSPR analysis, many analyses are still carried out incorrectly, or in a less than satisfactory manner. We have identified 21 types of error that continue to be perpetrated in the QSAR/QSPR literature, and each of these is discussed, with examples (including some of our own). Where appropriate, we make recommendations for avoiding errors and for improving and enhancing QSAR/QSPR analyses. PMID- 19544192 TI - Estimation of molecular diffusivity of pure chemicals in water: a quantitative structure-property relationship study. AB - A quantitative structure-property relationship (QSPR) study was performed to predict the molecular diffusivity of pure chemicals in water. A genetic-algorithm based multivariate linear regression (GA-MLR) was applied to select the most statistically effective molecular descriptors for modelling the molecular diffusivity of pure chemicals in water. Based on the selected molecular descriptors, a three-layer feed forward neural network (FFNN) was constructed to predict the property. The obtained results showed that the FFNN was able to predict the molecular diffusivity of pure chemicals in water. PMID- 19544193 TI - DFT study on the bromination pattern dependence of electronic properties and their validity in quantitative structure-activity relationships of polybrominated diphenyl ethers. AB - With quantum chemical computation of density functional theory (DFT), the electronic properties including the polarisabilities, polarisability anisotropies and quadrupole moments of a total of 209 congeners of polybrominated diphenyl ethers (PBDEs) were evaluated. The electronic properties were shown to be highly dependent on the bromination pattern, i.e. their values changed sensitively with the number and sites of bromination. Being similar to the 2,3,7,8-, 1,4,6,9 chlorination of dioxins, respectively, 3,3',4,4'-, 2,2',5,5'-bromination of PBDEs can impose relatively greater effects on the electronic properties. Some of electronic properties were found to be potent in explaining the variance of toxicity, and the potency was verified by the development of quantitative structure-activity relationships (QSARs). To further improve the stability and predictability of QSARs for toxicity, two-dimensional topological indices were introduced. In QSARs, polarisability anisotropy was more significant than other polarisability tensors, indicating the implicit occurrence of dispersion interaction between the ligand and aryl hydrocarbon receptor (AhR). For PBDEs, the quadrupole moment was as significant as shown previously for dioxins. As interesting descriptors with encoded information about dispersion and electronics, the electronic properties analysed herein are helpful in obtaining a better understanding of the congener-specific toxicities of PBDEs, and are applicable and may be extended to research into the toxicology of structurally similar compounds, such as halogenated aromatics. PMID- 19544195 TI - Application of the linear interaction energy method for rational design of artemisinin analogues as haeme polymerisation inhibitors. AB - The anti-malarial activity of artemisinin-derived drugs appears to be mediated by an interaction of the drug's endoperoxide bridge with intra-parasitic haeme. The binding affinity of artemisinin analogues with haeme were computed using linear interaction energy with a surface generalised Born (LIE-SGB) continuum solvation model. Low levels of root mean square error (0.348 and 0.415 kcal/mol) as well as significant correlation coefficients (r(2) = 0.868 and 0.892) between the experimental and predicted free energy of binding (FEB) based on molecular dynamics and hybrid Monte Carlo sampling techniques establish the SGB-LIE method as an efficient tool for generating more potent inhibitors of haeme polymerisation inhibition. PMID- 19544194 TI - QSAR models for P450 (2D6) substrate activity. AB - Human Cytochrome P450 (CYP) is a large group of enzymes that possess an essential function in metabolising different exogenous and endogenous compounds. Humans have more than 50 different genes encoding CYP enzymes, among these a gene encoding for the CYP isoenzyme 2D6, a CYP able to metabolise drugs and other chemicals. A training set of 747 chemicals primarily based on in vivo human data for the CYP isoenzyme 2D6 was collected from the literature. QSAR models focusing on substrate/non-substrate activity were constructed by the use of MultiCASE, Leadscope and MDL quantitative structure-activity relationship (QSAR) modelling systems. They cross validated (leave-groups-out) with concordances of 71%, 81% and 82%, respectively. Discrete organic European Inventory of Existing Commercial Chemical Substances (EINECS) chemicals were screened to predict an approximate percentage of CYP 2D6 substrates. These chemicals are potentially present in the environment. The biological importance of the CYP 2D6 and the use of the software mentioned above were discussed. PMID- 19544196 TI - Design of topological indices: computer-oriented approach. AB - A novel method is suggested for constructing topological indices (TIs) of molecular graphs which models human logic. This method is described in terms of a block scheme, consisting of the mutually connected elementary blocks. In each block the simple transformations of a molecular graph are fulfilled. A variant of the transformation is selected from the list of possible variants. Every TI is obtained as a result of the sequential execution of a number of operations, corresponding to some 'walk' on the block scheme. This walk can be selected both randomly and by the investigator. The suggested method can serve as a basis for the development of the respective computer program which may be used for the automatic construction of any number of TIs of differing nature. By this process one can also obtain the TIs that are unlikely to be constructed manually, due to their complexity. The set of obtained TIs may be used for building the structure property models. In the case of an unsatisfactory result the obtained set of TIs may be changed using the described generator of TIs. A number of examples of application of the suggested approach for the building QSAR/QSPR models is given. PMID- 19544197 TI - Use of principal component analysis and a spectral mapping technique for the evaluation of the antifungal activity of anthracene-based synthetic dyes. AB - The antifungal activity of 14 anthracene-based synthetic dyes and 6 reference compounds was measured on 36 fungal strains and the data matrix was evaluated separately by principal component analysis (PCA) and using a spectral mapping technique (SPM). The dimensionality of the maps of principal component loadings and variables and the selectivity maps was reduced to two by non-linear mapping. Except for two compounds, the dyes showed marked antifungal activity. Calculations proved that both the strength and selectivity of the biological effect of anthracene-based dyes were highly dependent on the chemical structure of the dye and on the type of fungi. PCA and SPM revealed different aspects of the antifungal activity, therefore, their simultaneous application in future quantitative structure-activity relationship studies is highly recommended. PMID- 19544198 TI - The physicochemical basis of QSARs for baseline toxicity. AB - The physico-chemical properties relevant to the equilibrium partitioning (bioconcentration) of chemicals between organisms and their respired media of water and air are reviewed and illustrated for chemicals that range in hydrophobicity. Relationships are then explored between freely dissolved external concentrations such as LC50s and chemical properties for one important toxicity mechanism, namely baseline toxicity or narcosis. The 'activity hypothesis' proposed by Ferguson in 1939 provides a coherent and compelling explanation for baseline toxicity of chemicals in both water- and air-respiring organisms, as well as a reference point for identifying more specific toxicity pathways. From inhalation studies with fish and rodents, narcosis is shown to occur at a chemical activity exceeding approximately 0.01 and there is no evidence of narcosis at activities less than 0.001. The activity hypothesis provides a framework for directly comparing the toxic potency of chemicals in both air- and water-breathing animals. The activity hypothesis is shown to be consistent with the critical body residue concept, but it has the advantage of avoiding the confounding effect of lipid content of the test organism. It also provides a theoretically sound basis for assessing the baseline toxicity of mixtures. It is suggested that since activity is readily calculated from fugacity, observed or predicted environmental abiotic and biotic fugacities can be used to evaluate the potential for baseline toxicity. Further, models employing fugacity or activity can be used to improve the experimental design of bioassays, thus possibly reducing unnecessary animal testing. PMID- 19544199 TI - The effect of predictive history on the learning of sub-sequence contingencies. AB - Two experiments demonstrated that the prior predictive history of a cue governs the extent to which that cue engages in sequence learning. Using a serial reaction time task, we manipulated the predictiveness of the stimulus locations (cues) with respect to the location of the stimulus on the next trial (outcome), such that half of the cues were good predictors of their outcomes, whilst the other half were poorer predictors. Following this, all cues were then paired with novel outcomes. Learning about those cues that were previously established as good predictors proceeded more rapidly than learning for those cues previously established as poor predictors. When the simple recurrent network is modified to include a variable associability parameter, the effects are easily modelled. PMID- 19544200 TI - Summer samples. PMID- 19544201 TI - Abnormal uterine artery Doppler in pregnancies suspected of a SGA fetus is related to increased risk of recurrence during next pregnancy. AB - OBJECTIVE: To investigate if placental Doppler velocimetry can predict the recurrence of a small-for-gestational age (SGA) fetus in subsequent pregnancies. DESIGN: Retrospective study. SETTING: City cohort over 15 years attending a university hospital. METHODS: A total of 196 pregnancies suspected of a SGA fetus (<3rd percentile) evaluated by uterine and umbilical artery Doppler velocimetry. Blood velocity waveform was analyzed for pulsatility index (PI) as well as the uterine artery waveform for notching in early diastole. MAIN OUTCOME MEASURE: The occurrence of a SGA newborn during the succeeding pregnancy by Doppler results from the previous pregnancy. RESULTS: In the group of 196 pregnancies suspected for SGA, 27 (13.8%) delivered a SGA newborn in the following pregnancy. Thirty seven (18.9%) of the 196 had an abnormally high PI in the uterine arteries in their first pregnancy, 12 (32.4%) of these delivered a SGA child in the next pregnancy (relative risk 3.44, p<0.001). The corresponding figure for those with normal uterine artery PI was 15 (9.4%). Abnormal umbilical artery Doppler was a worse predictor of recurrence of SGA (p=0.051). Uterine artery notching was not related to a SGA newborn during next pregnancy. CONCLUSION: The results suggest that abnormal uterine artery Doppler is related to increased risk of recurrence of SGA during the next pregnancy. This knowledge might provide the clinician with an opportunity to initiate preventive treatment and increase surveillance to women at risk during succeeding pregnancy. PMID- 19544202 TI - First trimester PTX3 levels in women who subsequently develop preeclampsia and fetal growth restriction. AB - Pentraxin 3 (PTX3) and C-reactive protein (CRP) levels were measured in the first trimester of pregnancy in women who subsequently developed preeclampsia (PE, n=16) and fetal growth restriction (FGR, n=12) requiring iatrogenic delivery before 37 weeks, and those who had uncomplicated pregnancies delivering at term (n=60). Mean PTX3 levels were significantly higher in women who subsequently developed PE (7.31 ng/ml, SD = 4.12) when compared to those with normal pregnancy outcome (4.92 ng/ml, SD = 1.94, p=0.0046). There were no significant differences between PTX3 levels in women with FGR (4.82 ng/ml, SD = 2.35) compared to normal pregnancy outcome (p=0.88). The median CRP levels did not vary significantly between the three groups (p=0.26). PTX3 levels in women who subsequently develop PE are already elevated in the first trimester, but not in those that develop FGR. This supports the hypothesis of an excessive maternal inflammatory response to pregnancy in the etiology of PE. PMID- 19544203 TI - Tooth loss prevalence and risk indicators in an isolated population of Brazil. AB - OBJECTIVE: The aim of this study was to assess the prevalence, extent, and risk indicators of tooth loss in an isolated population of Brazil. MATERIAL AND METHODS: Two-hundred-and-forty-two subjects, ranging in age from 14 to 82 years (mean 36.2 years), were identified by census in an isolated population of Brazil. All consenting subjects received a full-mouth clinical (DFT index and information about missing teeth) and periodontal examination of 6 sites per tooth. Furthermore, they were interviewed using a structured written questionnaire in order to gather information about demographic, environmental, and biological variables. RESULTS: Of the 200 subjects (80% response rate), 19 (9.5%) were edentulous, 90% had lost at least one tooth, and 39% had lost more than 8 teeth. The mean number of teeth lost was 9.5 (95% CI = 8.2-10.8). First mandibular molars were the most commonly missing teeth. In a multiple logistic regression analysis based on a theoretical hierarchical model of tooth loss, having more than 8 teeth lost was strongly associated with adult age (OR = 18.3-17.3, 95% CIs = 4.8-69.7 and 4.0-75.1) and female gender (OR = 5.9, 95% CI = 1.9-18.2) in the final model. CONCLUSIONS: Tooth loss was highly prevalent and extensive in this isolated population. Demographic and behavioral factors played an important role in tooth loss prevalence in this population. PMID- 19544204 TI - Indication for elective neck dissection for N0 carcinoma of the parotid gland: a single institution's 20-year experience. AB - CONCLUSIONS: Elective neck dissection (END) may not be necessary in patients with low-grade malignancy. However, currently, END should be performed in all patients with parotid carcinoma because the preoperative diagnosis of lymph node metastasis is limited and the accuracy of preoperative grade diagnosis is low. OBJECTIVE: Our aim was to examine the indication for END in N0 patients and the extent of dissection by investigating the accuracy of diagnosing the grade and lymph node metastasis of parotid carcinoma. PATIENTS AND METHODS: The subjects were 77 patients with parotid carcinoma. Among N0 patients, modified radical neck dissection (MRND) was conducted in those in whom the histological grade was evaluated as high, and selective neck dissection (SND) in other patients. RESULTS: Lymph node metastasis was detected in 27 of 71 patients who underwent neck dissection. Node metastasis was detected in 19 (61.3%) of 31 patients in whom high-grade malignancy was suggested and in 8 (22.9%) of 35 patients in whom intermediate-grade malignancy was suggested preoperatively. There was no metastasis in any of the five patients in whom low-grade malignancy was suggested. Occult metastasis was noted in 8 of 51 patients. The rate at which the histological grade was accurately diagnosed before surgery was low, especially in patients with low-grade malignancy. PMID- 19544205 TI - Toll-like receptor 2 gene polymorphism and its relationship with SCS in dairy cattle. AB - Toll-like receptor 2 (TLR2) plays an important role in the innate immune response to a variety of pathogens. In this study, bovine TLR2 gene was taken as a candidate gene for mastitis resistance. Through PCR-SSCP analysis and sequencing, three missense mutations at T385 G, G398A, and G1884A were detected in the coding region that encoded extracellular domain. Altogether 240 dairy cattle of three breeds (Holstein, Simmental, and Sanhe cattle) were genotyped and allele frequencies were determined. The effects of TLR2 polymorphisms on somatic cell score (SCS) were analyzed and significant association was found between T385 G and SCS. The mean of genotype GG was significantly lower than those of genotype TT and TG. No significant associations were found with SCS for G398A and G1884A. Information provided in this research will be useful in further studies to determine the role of TLR2 gene in the mastitis resistance. PMID- 19544206 TI - Exploratory study on the transcriptional profile of pigs subclinically infected with porcine circovirus type 2. AB - The aim of this exploratory work was to use a microarray-based approach to study the global transcriptome profile of caesarean-derived, colostrum-deprived (CDCD) piglets experimentally infected with porcine circovirus type 2 (PCV2). PCV2 inoculated piglets developed a subclinical infection, as confirmed by serology, in situ hybridization and quantitative PCR. Total RNA from mesenteric lymph nodes and lungs was obtained by duplicate from 2 control and 2 PCV2-inoculated piglets and was hybridized to Affymetrix Porcine GeneChip. Among the 24,123 probesets studied, 25 and 33 were found to be significantly differentially expressed (DE) between control and PCV2 groups for mesenteric lymph node and lung, respectively. Most up-regulated genes in PCV2 group were closely related to the immune response, such as cytokines (CCL4L, CXCL9, CXCL11), MHC binding molecules (TCRalpha, CD8alpha), immunoglubulins (IgG) and T cell activation (LCK, KLRK1, RASSF2, GBP2). Quantitative real-time PCR was performed to verify the microarray results. Therefore, from a transcriptional point of view, PCV2-inoculated pigs were apparently able to activate a cell-mediated response and develop PCV2 specific antibodies, which probably led to a subclinical infection. The results from this study indicate that a microarray based approach is a helpful tool in order to better understand the pathogenesis of PCV2 infection. PMID- 19544207 TI - Nucleotide sequence and association analysis of pig apolipoprotein-B and LDL receptor genes. AB - Three genes are the major determinants of heritable hypercholesterolemia diseases in humans: APOB, LDLR and LDLRAP1, which encode for proteins that physically interact to promote cholesterol uptake in the cell. We have carried out association analyses of these variants with serum cholesterol and triglycerides concentrations in a half-sib Duroc pig population. Given the structure of the population (six paternal half-sib families), we have used a statistical model that considers separately the allele transmission through dams (at population level) and through sires (within-families from heterozygous sire). Only polymorphisms showing a relevant substitution effect for both male- and female transmitted alleles are likely to be causal mutations. Thus, although we have found statistical association between genotypes for LDLR and APOB polymorphisms and serum lipid levels (mean allele substitution effects ranging from 15 to 40% of the standard deviation of these traits), none of them seem to be the causal mutation but probably represent closely linked polymorphisms. We have shown here that these three genes also contribute to genetic variability in pigs, with the description of new polymorphisms in their coding regions. Moreover, we have demonstrated that variants on two of these three genes are segregating in a number of commercial breeds. Finally, we report here the coding region for the porcine LDLRAP1 gene and describe a polymorphism in the last exon of this gene. PMID- 19544208 TI - PLGA microsphere-mediated growth hormone release hormone expression induces intergenerational growth. AB - To improve animal growth, growth hormone-releasing hormone (GHRH) expression vectors that maintain constant GHRH expression can be directly injected into muscles. To deliver the GHRH expression vectors, biodegradable microspheres have been used as a sustained release system. Although administering GHRH through microspheres is a common practice, the intergenerational effects of this delivery system are unknown. To investigate the intergenerational effects of polylactic-co glycolic acid (PLGA) encapsulated plasmid-mediated GHRH supplements, pCMV-Rep GHRH microspheres were injected into pregnant mice. Growth and expression of GHRH were measured in the offspring. RT-PCR and immunohistochemistry reveal GHRH expression 3-21 days post-injection. The proportion of GH-positive cells in the GHRH treated offspring was 48.2% higher than in the control group (P < 0.01). The GHRH treated offspring were 6.15% (P < 0.05) larger than the control offspring. At day 49 post-injection, IGF-I serum levels were significantly higher in the treatment group than in the control group. This study confirms that intramuscular expression of GHRH mediated by PLGA microspheres significantly enhances intergenerational growth. PMID- 19544209 TI - Cloning and spatio-temporal expression of porcine CDK5 and CDK5R1(p35) genes. AB - Cyclin-dependent kinase 5 (CDK5) is a serine/threonine kinase homologue attributed to the mitotic cyclin-dependent kinase family. Both the kinase activity and the biological effects of CDK5 in central nervous system are mainly dependent on association with its regulatory subunit 1 known as CDK5R1 (p35). In the present study, the full-length coding regions of CDK5 and CDK5R1 were cloned from pigs. Radiation hybrid mapping localized porcine CDK5 to chromosome 18q12 13, whereas CDK5R1 was electro-localized to chromosome 12q12. Real-time quantitative RT-PCR (qRT-PCR) showed that CDK5 mRNA is ubiquitously present in all porcine tissues examined, with relatively high levels in cerebral cortex, cerebellum, testicle and lung. We also examined the expression profile of porcine CDK5/CDK5R1 in various tissues at different developmental stages. The results indicated that CDK5 mRNA reaches the highest level in cerebral cortex at two months of age and in cerebellum and liver at 4 months of age, respectively, whereas the peak level of CDK5R1 was observed in both cerebral cortex and cerebellum at two months of age, indicating the pivotal role of CDK5/CDK5R1 during the development of porcine brain. PMID- 19544210 TI - Somatostatin can alter fertility genes expression, oocytes maturation, and embryo development in cattle. AB - This study was designed to investigate the effect of somatostatin on oocytes maturation and subsequent embryo development in cattle. Bovine granulosa cells separated from oocytes, cultured for 24 h and transfected with pEGFP-N1 vector with mouse SST gene (Experimental) and with out plasmid transfection (Control). RT-PCR and Real-Time PCR were used to estimate the expression of bovine receptors of androgen, estrogen beta, growth hormone, and follicular stimulating hormone. Culture media concentrations of hormones were measured by kits using radioimmunoassay. COCs aspirated from ovaries were co-cultured with granulosa cells layers (transfected or control) at 38.5 degrees C in CO(2) incubator for maturation. We found a significant (2.37X) increase in estrogen receptor beta expression in experimental group. There was a decrease in androgen receptor, growth hormone releasing hormone receptor, and follicular stimulating hormone receptor (P < 0.05). But, 96 h of post transfection, culture media concentration of estradiol-17beta was increased significantly (P < 0.05) and testosterone, growth hormone and follicular stimulating hormone showed opposite trend (P < 0.05) in experimental groups. Co-culture of somatostatin transfected granulosa cells with oocytes, reduced the maturation rate from 70% to 66% but had no effect on subsequent fertilization and embryo development. PMID- 19544211 TI - Variable levels of 37-kDa/67-kDa laminin receptor (RPSA) mRNA in ovine tissues: potential contribution to the regulatory processes of PrPSc propagation? AB - The 37-kDa laminin receptor precursor/67-kDa laminin receptor (LRP/LR, also known as ribosomal protein SA, RPSA) has been reported to be involved in cancer development and prion internalization. Previous studies have shown that the LRP/LR is expressed in a wide variety of tissues. In particular, expression of LRP/LR mRNA may be closely related to the degree of PrP(Sc) propagation. This study presents a detailed investigation of the LRP/LR mRNA expression levels in eleven normal ovine tissues. Using real-time quantitative PCR, the highest LRP/LR expression was found in neocortex (p < 0.05). Slightly lower levels were found in the heart and obex. Intermediate levels were seen in hippocampus, cerebellum, spleen, thalamus, mesenteric lymph node, and the lowest levels were present in liver, kidney, and lung. In general, the LRP/LR mRNA levels were much higher in neuronal tissues than in peripheral tissues. The observation that differences in LRP/LR mRNA expression levels are consistent with the corresponding variation in PrP(Sc) accumulation suggests that the 37-kDa/67-kDa laminin receptor may be involved in the regulation of PrP(Sc) propagation. PMID- 19544212 TI - Identification of novel allelic variants of integrin beta 2 (ITGB2) gene and screening for Bubaline leukocyte adhesion deficiency syndrome in Indian water buffaloes (Bubalus bubalis). AB - A fragment of 570 bp corresponding to exon 5 and 6 of integrin beta 2 (ITGB2) gene was amplified for screening D128G mutation in one hundred and fifty two buffaloes (Bubalus bubalis) which causes bovine leukocyte adhesion deficiency syndrome (BLAD) in cattle, as well as to ascertain polymorphism. TaqI PCR-RFLP revealed no such mutation thus indicating the absence of bubaline leukocyte adhesion deficiency (BuLAD) allele in animals under study. However, the polymorphism studies using MspI restriction enzyme revealed two genotypic patterns viz. AA pattern (bands of 293, 141, 105, and 31 bp) and BB pattern (bands of 293, 105, 77, 64, and 31 bp). The sequences of A and B alleles were submitted to the GenBank (EU853307 and AY821799). PMID- 19544213 TI - New single nucleotide polymorphisms in the mu-calpain gene in Spanish maternal beef breeds. AB - Calpains play an important role in the postmortem tenderization process of meat and several SNP in the mu-calpain gene (CAPN1) have been reported to be associated with tenderness in beef cattle. Our objectives were to identify the previously reported CAPN1 331G>C SNP and to detect new polymorphisms in this gene in Spanish maternal beef breeds. A fragment (exon 8 to 10) of the bovine CAPN1 gene was sequenced and genotyped in a sample of the main Spanish maternal beef breeds including Retinta, Morucha, and Avilena Negra-Iberica. These breeds are characterized for their high meat quality, their adaptation to adverse environmental conditions, and their good maternal aptitude. This adaptation makes it possible to rear these breeds in the south and west of Spain, where drought and feed shortages occur frequently. Six SNP in the mu-calpain gene were found, five of which (CAPN1 80C>T, 302C>G, 310G>A, 445C>T, 524A>C) have not been reported previously. Sequences obtained for these five newly found SNP were submitted to GenBank (Accessions EU386166 to EU386183). PMID- 19544214 TI - Molecular cloning and characterization of cDNA encoding buffalo (Bubalus bubalis) cytochrome P450 aromatase in the ovary. AB - One of the predominant causes of poor reproduction in buffaloes is low levels of ovarian estrogens. A rate limiting enzyme in estrogen biosynthesis is cytochrome P450 aromatase (P450 AROM), the product of CYP19 gene. In the present study CYP19 cDNA was cloned and its 5'UTR was characterized by 5'RACE in granulosa cells of large follicles. CYP19 transcripts with four different 5'UTRs (206, 114, 90 and 3 bases) were found in buffalo granulosa cells of large ovarian follicles. Interestingly, a predominant aromatase transcript with short 5'UTR (3 nucleotides) was found. Further studies are required to understand the relevance of these transcripts and their translational efficiency in granulosa cells of large follicles during folliculogenesis of buffalo ovary. PMID- 19544215 TI - Association between sexual precocity and alleles of KISS-1 and GPR54 genes in goats. AB - KISS-1 and GPR54 were regarded as key regulators for the puberty onset and fundamental gatekeepers of sexual maturation in mammals. To explore the possible association between variations in KISS-1 and GPR54 with sexual precocity, mutation screening of exon 1 of KISS-1 and exon 1, exon 3, and partial exon 5 of GPR54 was performed in a sexual precocious breed (Jining Grey goats) and sexual late-maturing breeds (Inner Mongolia Cashmere, Angora, and Boer goats) by PCR SSCP. The results showed that five novel mutations were identified in exon 1 and partial exon 5 of GPR54 including C96 T, T173C, G176A, G825A, and C981 T. The Jining Grey goats with genotype BB or AB had 1.07 (P < 0.05) or 0.40 (P < 0.05) kids more than those with AA. The Jining Grey goats with genotype DD or CD had 1.80 (P < 0.05) or 0.55 (P < 0.05) kids more than CC, respectively. The present study preliminarily showed an association between alleles B and D of GPR54 with high litter size and sexual precocity in Jining Grey goats. PMID- 19544216 TI - Men sexually interested in transwomen (MSTW): gendered embodiment and the construction of sexual desire. AB - This article extends research on transgenderism by providing a sociological study of men who are sexually interested in transwomen (MSTW; viz., genetic males who use estrogen to feminize their body but retain their penis). We conducted fieldwork in a bar catering to transwomen and the men who were sexually interested in them, and did on-the-spot interviews with the MSTW. We initially examined the nature of MSTW's sexual attraction to transwomen, followed by how this related to their sexual orientation identity. Using a sexual fields approach (Green, 2008), we first found how the ambience of the bar helped to create an erotic environment through a heightened sense of gender. Then, focusing on the theory of embodiment (Jackson & Scott, 2007), we saw how the MSTW constructed a unique sexual desire according to the sexual orientation identity they brought to the situation. Those who identified as "straight" tended to gloss that the transwoman had a penis, while the bisexually identified men were more likely to incorporate the transwoman's penis into the sexual experience. PMID- 19544217 TI - CT-guided robotically-assisted infiltration of foot and ankle joints. AB - It was our aim to describe a CT-guided robotically-assisted infiltration technique for diagnostic injections in foot and ankle orthopaedics. CT-guided mechatronically-assisted joint infiltration was performed on 16 patients referred to the orthopaedic department for diagnostic foot and ankle assessment. All interventions were performed using an INNOMOTION-assistance device on a multislice CT scanner in an image-guided therapy suite. Successful infiltration was defined as CT localization of contrast media in the target joint. Additionally, pre- and post-interventional VAS pain scores were assessed. All injections (16/16 joints) were technically successful. Contrast media deposit was documented in all targeted joints. Significant relief of pain was noted by all 16 patients (p<0.01). CT-guided robotically-assisted intervention is an exact, reliable and safe application method for diagnostic infiltration of midfoot and hindfoot joints. The high accuracy and feasibility in a clinical environment make it a viable alternative to the commonly used fluoroscopic-guided procedures. PMID- 19544218 TI - A new thoracoscopic technique for pulmonary lobectomies. AB - Video-assisted thoracoscopic surgery (VATS) for pulmonary lobectomies has recently been introduced as a minimally invasive technique for selected patients. Herein I describe an original technique, which allows for lobectomies under the direct vision of a surgeon respecting the principles that are the basis of the thoracoscopic techniques. The originality of the technique consists in the supine position of the patient and the use of a modified retractor for lower lobectomies. This approach required two ports, an access incision (6-7 cm), and an additional port which may be used for lower lobectomies. Utilizing this technique I operated on twenty patients affected by stage I lung cancer with poor respiratory function. The ability to perform lobectomies with this method allows for shorter operative time than when utilizing previous techniques (80 min. mean operative time) including the lymph nodal dissection. Moreover, cosmetic results were good and patients benefited from excellent early pulmonary function and fast recovery time. I recommend the use of this approach which is a safe technique for the patient and an advantageous procedure for both surgeon and anaesthesiologist. PMID- 19544219 TI - Childhood trauma and dissociation in women with pseudoseizure-type conversion disorder. AB - BACKGROUND: Conversion disorder is thought to be associated with psychological factors because of the presence of conflict and other stressors prior to the condition. AIM: The aim of this study is to compare adult patients with pseudoseizure-type conversion disorder with healthy control group in terms of childhood trauma, dissociative disorder and family history of psychiatric disorders. METHOD: 56 female patients were admitted to the general psychiatry hospital outpatient clinic between January and July 2005. All patients had a negative experience about their families just before having the conversion. Diagnosis was made according to the DSM-IV criteria. A control group consisting of similar patient demographics of the disease group has been selected. Socio demographic information forms, the Childhood Trauma Questionnaire (CTQ) and Dissociation Questionnaire (DIS-Q), were completed on the patients. RESULTS: CTQ total (t=12.12, P<0.001) and subscales, emotional abuse and emotional neglect (EA EN) (t=12.74, P<0.001), physical abuse (PA) (t=10.05, P<0.001), and sexual abuse (SA) (t=7.69, P<0.001) were significantly high in the conversion group. DIS-Q mean points were statistically higher in the conversion group (t=11.05, P<0.001). CONCLUSIONS: The findings suggest that pseudoseizures (conversion disorder) should be included within dissociative disorders in DSM system as in ICD. It is usually uncommon for the patient to tell about childhood trauma without being specially questioned about this issue. Thus, it would be helpful to uncover these experiences by using related scales in conversion disorder patients. PMID- 19544220 TI - Acute homing of bone marrow-derived mononuclear cells in intramyocardial vs. intracoronary transplantation. AB - OBJECTIVES: Cell homing optimisation after transplantation is critical in myocardial infarction (MI) cell therapy. DESIGN: Eight pigs were randomized to receiving autologous purified (111)indium-labeled bone marrow mononuclear cells (BMMCs) (10(8) cells/2 ml) by intramyocardial (IM) (n=4) or by intracoronary (IC) (n=4) transplantation after 90 minutes occlusion of the CX-coronary artery. Dual isotope SPECT imaging was performed 2 and 24 hours postoperatively. Two animals were additionally analyzed on the sixth postoperative day. Tissue samples from the major organs were analyzed. RESULTS: In SPECT imaging revealed that BMMCs administered using IM injection remained in the injured area. In contrast, minor proportion of IC transplanted cells remained in the myocardium, as most of the cells showed homing in the lungs. Analysis of the biopsies showed a seven-fold greater number of cells in the myocardium for the IM method and a 10-fold greater number of cells in the lungs in the IC group (p < 0.001). CONCLUSIONS: In producing persistently high cell homing at the infarction site, the IM transplantation is superior to the IC transplantation. However, the IC administration might be more specific in targeting injured capillaries and epithelial cells within the infarcted myocardium. PMID- 19544221 TI - A single center experience with the HeartMate II left ventricular assist device (LVAD). AB - OBJECTIVES: Left ventricular assist devices (LVAD), used in the setting bridge-to transplantation and destination therapy, for patients with deteriorating severe heart failure are continuously developing. The second generation, the axial flow pumps, have been introduced since some years. DESIGN: Eleven consecutive patients, seven male, with severe heart failure due to ischemic cardiomyopathy (n=5), dilated cardiomyopathy (n=5) and cytotoxic etiology (n=1) were implanted with the HeartMate-II. They were preoperatively treated with inotropic support (n=9), ventricular assist device (n=2) and mechanical ventilation (n=4). RESULTS: Eight patients were bridged to transplant after median 155 days (range, 65 to 316 days). One patient is ongoing for 748 days, intended for destination therapy. Ten of eleven patients were discharged after median 64 days (range, 40 to 105 days). Four patients were reoperated due to bleeding. Two embolic events were recorded. One perioperative death. CONCLUSION: Eleven HM-II LVAD's have been implanted in our institution with good early results. Eight patients were successfully bridged to heart transplantation. One patient is intended for destination therapy and is ongoing since November 2006. In these severely ill patients, this technique offers a good chance surviving until heart transplantation. In selected cases the technique also offers the possibility of a permanent support and longevity. PMID- 19544222 TI - The influence of genotype on vascular endothelial growth factor and regulation of myocardial collateral blood flow in patients with acute and chronic coronary heart disease. AB - OBJECTIVE: To test the hypothesis that mutations in the vascular endothelial growth factor (VEGF) gene are associated with plasma concentration of VEGF and subsequently the ability to influence coronary collateral arteries in patients with coronary heart disease (CHD). METHODS: Blood samples from patients with chronic ischemic heart disease (n=53) and acute coronary syndrome (n=61) were analysed. Coronary collaterals were scored from diagnostic biplane coronary angiograms. RESULTS: The plasma concentration of VEGF was increased in patients with acute compared to chronic CHD (p=0.01). The genotype frequencies differed significantly from Hardy-Weinberg equilibrium in three of 15 examined loci. Four new mutations in addition to the already described were identified. The VEGF haplotype did not seem to predict plasma VEGF concentration (p=0.5). There was an association between the genotype in locus VEGF-1154 and coronary collateral size (p=0.03) and a significant association between the VEGF plasma concentration and the collateral size (p=0.03). CONCLUSION: VEGF plasma concentration seems related to coronary collateral function in patients with CHD. The results did not support the hypothesis that polymorphisms in the untranslated region of the VEGF gene were associated with the concentration of circulating VEGF. Increased understanding of VEGF in the regulation of myocardial collateral flow may lead to new therapies in CHD. PMID- 19544223 TI - A simple and objective marker for stress. AB - OBJECTIVE: To examine the association between pressure pain sensitivity (PPS) at sternum and various well established physiological stress measures among opera singers during a performance as a measure for transitional stress, and resting values in out-clinic patients as a measure for persistent stress. METHODS: Changes in PPS on the index finger and sternum, middle blood pressure (MAP), heart rate (HR), pressure-rate-product (PRP) and salivary cortisol (SCO) were recorded in 26 opera solo singers during a performance. Resting PPS, HR, MAP, PRP and presence of a noxious withdrawal reflex (NWR) were recorded in 181 out-clinic patients. RESULTS: During opera performance, the PPS on sternum changed concomitantly with MAP (correlation coefficient (r) r=0.42, p<0.005), HR (r=0.55, p<0.001), PRP (r=0.54, p<0.001) and SCO (r=0.26, p=0.066). During rest, a significant correlation was found between PPS on sternum and HR, PRP and presence of noxious withdrawal reflex (all p<0.01). CONCLUSIONS: The PPS measurement at sternum was associated with well established physiological stress measures and may represent a simple, objective and reliable measure of physiological stress used by both non-professional and professionals. PMID- 19544225 TI - Frequency of norovirus in stool samples from hospitalized children due to acute gastroenteritis in Anatolia, Turkey, 2006-2007. AB - Noroviruses are among the most common causes of sporadic enteritis in childhood. In this pilot study, the frequency of norovirus infection in children in mid western Turkey was investigated from November 2006 to June 2007. Noroviruses were detected in 17% of samples (15/88) by a combination of 2 different RT-PCR assays, both targeting an overlapping region of the RNA-dependent RNA polymerase gene. By sequence analysis, most strains were characterized as GIIb/Hilversum. One strain was characterized as GII.4/2006a, a variant that appeared worldwide in 2006, while another strain was characterized as a rare genotype, GII.6. This study demonstrates the importance of norovirus in paediatric diarrhoea and suggests the heterogeneity of circulating strains in Turkey. PMID- 19544224 TI - Entecavir to treat severe acute hepatitis B. PMID- 19544226 TI - Disinhibition of sequential actions following right frontal lobe damage. AB - Action disorganization syndrome (ADS) is a frontal lobe neuropsychological syndrome characterized by deficits in the performance of familiar sequential tasks such as making a cup of tea. In this study, we examined the performance of familiar sequential tasks by 3 patients with right frontal brain tumours and no other areas of brain damage. When task-irrelevant objects were presented as distractors in addition to the target objects, all patients used the distractors in accordance with the target task. These patients showed few of the sequential and omission errors that had been previously reported in patients with ADS. Although normal participants could suppress the activation of task-irrelevant objects, these patients had difficulty with this process, which would normally be carried out by the right frontal lobe to determine the most suitable action behaviour. The intact left frontal lobe may receive bottom-up activation from the distractors and modify this schema to match the behavioural context. Our findings suggest that patients with only right frontal lobe damage may be characterized more by action disinhibition than by disorganization. PMID- 19544227 TI - ACTN3 R577X polymorphism and Israeli top-level athletes. AB - A common genetic variation in the alpha-actinin-3 ( ACTN3) gene causes a replacement of an arginine (R) with a premature stop codon (X) at amino-acid 577 (rs1815739). While the R allele has been found to be associated with power oriented performance, the XX genotype may be linked with endurance ability. To test this hypothesis, we studied the distribution of ACTN3 genotypes in 155 Israeli athletes (age=35.9+12.2 years) classified by sport (endurance runners and sprinters) and in 240 sedentary individuals. The sprinters' allele frequencies (AF: R/X=0.7/0.3) and 577RR genotype distribution percentage (GD: RR=52%) differed markedly from those of the endurance athletes (AF: R/X=0.53/0.47, p=0.000007; GD: RR=18%, p=0.00009) and the control group (AF: R/X=0.55/0.45, p=0.0002; GD: RR=27.3%, p=0.000003). A comparison between the top-level and national-level sprinters revealed that the R allele occurs more often in the top level sprinters. A significantly higher proportion of the XX genotype was observed in endurance athletes (34%) compared with controls (18%, p=0.02) and sprinters (13%, p=0.002). However, top-level and national level endurance athletes had similar allele and genotype frequencies. We conclude that the ACTN3 R allele is associated with top-level sprint performance and the X allele and XX genotypes may not be critical but rather additive to endurance performance. PMID- 19544228 TI - Cystatin C a marker for renal function after exercise. AB - Renal impairment is common during and after severe exercise. In clinical practice, renal function is evaluated using serum creatinine, urine parameters, and equations to estimate the Glomular Filtration Rate (GFR). However, creatinine levels may be biased by skeletal muscle damage and the GFR equations, requiring age, gender and body weight, are shown to be inadequate in normals. In the present study, we show that serum cystatin C and creatinine concentrations were elevated after marathon running in 26% and 46% of the 70 recreational male runners, respectively, possibly because of reduction in renal blood flow. The mean cystatin C increase was twice as low as compared to creatinine (21% and 41%, respectively), suggesting that cystatin C is indeed less biased by muscle damage. Future research has to reveal whether training diminishes the elevation in renal markers. Overall, cystatin C seems a more reliable method to establish renal function during and after extensive exercise. PMID- 19544229 TI - [Follow-up of children with prenatal diagnosis of supraventricular extrasystole]. AB - PURPOSE: The aim of this study was the follow-up of children with a prenatal diagnosis of supraventricular extrasystole (SVES) up to an age of 5 years in order to assess the long term outcome of these children and in order to characterize factors influencing the outcome. MATERIALS AND METHOD: All fetuses diagnosed with prenatal SVES between April 1993 and August 2005 were identified and the children's parents and pediatricians contacted for data regarding the children's health. Follow-up data about the children's health up to the age of 5 years could be obtained in 77 (46.1%) children. RESULTS: 0.5% (167/34,770) of all fetuses were diagnosed with prenatal SVES. In 70% of cases the SVES resolved before or at birth. 30% of children presented with arrhythmia postpartum. 31% of children were diagnosed with cardiac anomalies postpartum. 87% of children were healthy at the age of 2 - 5 years. In the subgroup of children with isolated fetal SVES without further anomalies, 95% of children were healthy at the age of 2 - 5 years. In children with persisting arrhythmia and in children with cardiac anomalies, the prognosis was worse. CONCLUSIONS: Overall, by the long-term follow up of 77 children with prenatal SVES we could show that prenatal SVES has a good prognosis. However, 30% of children develop arrhythmia postpartum and 31% of children present with cardiac anomalies. These children still have a worse prognosis than children with isolated prenatal SVES. PMID- 19544230 TI - Transthoracic ultrasonography-guided core needle biopsy of pleural-based lung lesions: prospective randomized comparison between a Tru-cut-type needle and a modified Menghini-type needle. AB - PURPOSE: The diagnostic yield of the different types of cutting needles used to perform transthoracic biopsy is scarcely investigated. Aim of the study was to compare a Tru-cut-type (TCT) needle and a modified Menghini-type needle (MMT) in ultrasonography (US)-guided biopsy of pulmonary lesions. MATERIALS AND METHODS: 307 subjects (191 males and 116 females, mean age 58 years) with peripheral lung lesions selected to undergo US-guided biopsy were randomized to undergo biopsy by using an 18-gauge TCT or MMT needle. The specimens were imprinted on two to three slides for cytology and then put into a formalin solution for histology. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and overall accuracy were calculated. Fisher's exact test was used to compare histology recovery rate (HRR), diagnostic accuracy, and diagnostic yield of the combination of cytology and histology in comparison with cytology alone and histology alone. RESULTS: 155 biopsies were performed using the MMT needle, 152 using the TCT needle. HRR was 112 / 155 (72.3 %) and 144 / 152 (94.7 %), respectively (p < 0.0001). Sensitivity, specificity, PPV, and NPV were 81.6 %, 100 %, 100 %, and 62 % for the former, respectively, and 93.6 %, 100 %, 100 %, and 86 % for the latter. A correct diagnosis was achieved in 133 / 155 biopsies (85.8 %) performed with the MMT needle, and in 145 / 152 biopsies (95.4 %) performed with the TCT needle (p = 0.0041). The combination of cytology and histology had a higher diagnostic yield than cytology alone (p < 0.001) and histology alone (p < 0.001). CONCLUSION: The TCT needle performs better than the MMT needle, and improves the diagnostic accuracy of US-guided transthoracic biopsy of superficial lung lesions. PMID- 19544231 TI - Measurement of common carotid artery intima-media thickness in clinical practice: comparison of B-mode and RF-based technique. AB - PURPOSE: The common carotid artery intima-media thickness (CCA-IMT) is usually measured using B-mode ultrasound images. A different approach for CCA-IMT detection is based on radio frequency (RF) multiple M-line analysis. MATERIALS AND METHODS: The present study explores the relationship between B-mode and RF measurement of CCA-IMT, as well as the reproducibility of both methods in 136 patients recently diagnosed with cardiovascular disease. Within one session, repeated measurements were made in the distal CCA bilaterally, using the B-mode (averaged over 10 mm) and RF technique (averaging 12 M-lines over 14 mm). RESULTS: The two methods correlate well (Pearson r = 0.765). The CCA-IMT values measured with B-mode and RF were 0.779 +/- 0.196 mm and 0.734 +/- 0.172 mm, respectively. B-mode CCA-IMT is significantly larger than RF CCA-IMT (mean difference of 0.045 mm, SEM 7.8 microm; t = 5.82; p < 0.001). In the multivariate regression analysis, carotid artery stenosis, inhomogeneous IMT and diabetes mellitus were the main predictors of differences between B-mode and RF CCA-IMT. The intrapatient variation for B-mode and RF-based CCA-IMT is comparable (0.05 +/ 0.04 mm and 0.07 +/- 0.05 mm, respectively). CONCLUSION: CCA-IMT values measured with RF and B-mode have similar reproducibility and exhibit acceptable correlation, but RF CCA-IMT is significantly smaller. The difference between both methods is mainly due to advanced atherosclerosis. Hence, both methods can be used reliably to measure CCA-IMT in clinical practice. PMID- 19544232 TI - Fetal weight estimation by 2D and 3D ultrasound: comparison of six formulas. AB - PURPOSE: To evaluate and compare the accuracy of different formulas to estimate fetal weight using 2D and 3D ultrasound. MATERIALS AND METHODS: We performed a prospective study on unselected singleton pregnancies. All scanned fetuses delivered within 7 days in absence of structural and chromosomal abnormalities were included. The fetal weight was calculated using the 3D Schild, Chang, Liang and 2D Hansmann, Merz and Hadlock formulas. Absolute and mean deviations of estimated fetal weight were calculated. RESULTS: Of 249 scanned fetuses 200 that fulfilled the criteria were included. Birth weights ranged between 535 and 5020 g. The highest correlation between estimated fetal and birth weight was achieved by applying Schild's equation, and the absolute percentage error was 5.6%. The corresponding values for 2D Hansmann, Merz and four-parametric Hadlock formulas were 7.5%, 7.9%, and 9.2%, respectively, while these were 13.1% and 30.7% for Liang's and Chang's 3D equations. Using the Schild formula, a deviation from birth weight below 10 % was achieved in 80 % of fetuses, with Hansmann's in 73.5% and with Merz in 72.5%, while this parameter was much lower in the remaining equations. CONCLUSION: The best option with the highest accuracy for sonographic fetal weight estimation was the 3D Schild equation followed by the 2D Hansmann and Merz formulas. Published data of the accuracy could be reproduced with the exception of the "Asian" 3D equations in our European population. The limited improvement in weight agreement using the 3D technique compared to the 2D technique may be outweighed by the time consumption. PMID- 19544233 TI - Ultrasound characteristics of breast fibroadenomas are related to clinical and histological parameters. AB - PURPOSE: We hypothesized that ultrasound characteristics of breast fibroadenomas (FA) vary in relation to the clinical and histological parameters: patient age, tumor size and histological classification. MATERIALS AND METHODS: Eleven ultrasound characteristics frequently observed in breast tumors were defined before the onset of our study. These characteristics, as well as a semi quantitative score for vascularization on color-coded Doppler ultrasound, were analyzed in a retrospective study. Histology revealed adult type differentiation in all FA. They were divided into florid, regressive and mixed subtypes. The examiner was blinded for the histological classification during image analysis. RESULTS: Histological type: florid FA: more frequent in younger women (age group < 30 years; p < 0.001), and bigger than regressive FA (larger than 16 mm: p = 0.007). Statistically significant differences between florid and regressive FA regarding the ultrasound features: enhanced posterior ultrasound transmission (p < 0.001), homogenous echo pattern (p = 0.003) and lobulated margin contour (p = 0.042). Tumor size: patients with larger tumors (> 16 mm) were younger (mean age 35 vs. 43 years, p < 0.001). More often in bigger FA: enhanced dorsal ultrasound transmission (p < 0.001), hyperechoic spots (p < 0.001), strong vascularization (p < 0.001), inhomogeneous echo pattern (p = 0.001), horizontal axis (p = 0.009), lobulated margin contour (p = 0.009), lateral shadowing (p = 0.047). Age: more often in older patients (age group > 30 years): dorsal ultrasound shadowing (p = 0.008), irregular margin contour (p = 0.038), homogenous echo pattern (p = 0.047). CONCLUSION: Histological type, tumor size and patient age significantly influence ultrasound characteristics of breast FA. This might be helpful to consider when breast lesions are classified and decisions for biopsies are made. PMID- 19544234 TI - [Dehiscence of the uterine scar after three previous caesarean sections]. PMID- 19544235 TI - [Detection and treatment monitoring of secondary coeliac trunk endoleak following hybrid open-endovascular aortic aneurysm repair using B-flow and contrast enhanced ultrasound]. PMID- 19544236 TI - [An new generation of antineoplastic agents--targeted therapy in gastrointestinal tumors]. PMID- 19544238 TI - [Thoracic outlet syndrome--complications (OLG Hamm 3. Civil court, decision of 18th April 2005, Az. 3 U 259/04--treatment error due to outdated operation method?]. PMID- 19544239 TI - [What properties are required for once becoming a good psychotherapist?]. PMID- 19544240 TI - [Strengths orientation]. PMID- 19544241 TI - [Obituary to Margit von Kerekjarto]. PMID- 19544242 TI - [Empirical validation of a model for the etiology of eating disorders in adolescent girls: which role play the results of body comparisons?]. AB - Aim of this study was the empirical validation of an etiologic model on disturbed eating behavior by means of structural equation modeling. The model under consideration by Schutz and colleagues explains the influence of potential risk factors, especially body comparisons, on disturbed eating. 136 female sixth graders from secondary schools in Thuringia filled out questionnaires on their eating behavior, on the variables stated in the model and on the subjective meaning of their body weight. The model is supported by our data analyses. Particularly BMI and self esteem have a strong influence on body comparisons. The impact of body comparisons on disturbed eating is mediated by body dissatisfaction. An important addition to the model can be seen in the inclusion of the subjective effect of body comparisons into the model which has been neglected in research so far. PMID- 19544243 TI - [Revalidation of a questionnaire assessing women's satisfaction with maternity care in hospital]. AB - AIM: Revalidation of a survey instrument assessing women's satisfaction with maternity care in hospital. To determine the suitability of the questionnaire for its use in internal and external quality management. METHODS: Postal survey, 3 073 women from 38 hospitals received a questionnaire after discharge. Psychometric evaluation was done via exploratory factor analysis and reliability analysis. The questionnaire's ability to differentiate between subgroups and between maternity wards was evaluated. RESULTS: The mean response rate was 58%. Eleven factors (Cronbach's Alpha between 0.7 and 0.9) were extracted, explaining 55% of the variance. Parity and getting to know the staff beforehand was associated with women's satisfaction especially with issues encompassing interaction with the staff. CONCLUSION: Due to its good psychometric properties, the ability to differentiate between hospitals and its practicability the questionnaire is well suited for use in internal and external quality management of maternity wards. PMID- 19544244 TI - [Screening for depersonalization-derealization with two items of the cambridge depersonalization scale]. AB - Depersonalization (DP) and derealization (DR) are considered to be highly underdiagnosed. Therefore the development of screening instruments is important. From the Cambridge Depersonalization Scale (CDS) two items were extracted discriminating best patients with clinical significant DP from patients without DP. These two Items were assembled to a short version of the CDS. This short version (CDS-2) was tested in a sample of 38 patients with clinical significant DP-DR and 49 patients without or only mild DP-DR. Scores were compared against clinical diagnoses based on a structured interview (gold standard). The CDS-2 was able to differentiate patients with clinical significant DP well from other groups (cut-off of CDS-2>or=3, sensitivity=78.9%, specifity=85.7%) and also showed high reliability (Cronbachs alpha=0.92). Therefore the CDS-2 can be considered as a useful tool for screening and identification of DP-DR. PMID- 19544245 TI - Altered hypothalamic-pituitary-adrenal axis activity in patients with chronic heart failure. AB - Neuroendocrine factors play an important role in the pathogenesis of chronic heart failure. Despite numerous clinical and experimental studies, the role of the hypothalamic-pituitary-adrenal axis and glucocorticoid hormones is not fully characterised. Here we present a study of plasma cortisol concentration in 74 chronic heart failure patients, divided into four groups based on NYHA functional classes I-IV, and in 17 control subjects. In parallel, we performed morphological analysis of the hypothalamic-pituitary-adrenal axis components from 8 male patients who had died from chronic heart failure, and 9 male controls. In our study we applied immunohistochemical method and quantitative analysis to investigate an expression of hypothalamic neurohormones (corticotropin-releasing hormone, vasopressin) and adrenocorticotropin hormone in the pituitary, as well as performed general histological examination of the adrenal cortex. Measurement of morning cortisol concentration in plasma of chronic heart failure patients revealed neither difference compared to controls nor with the severity of the disease. Despite this, a two-fold increase in the density of corticotropin releasing hormone-immunoreactive neurons as well as a two-fold increase in the number of corticotropin-releasing hormone neurons co-expressing vasopressin in the hypothalamic paraventricular nucleus were found. In the anterior pituitary the density of adrenocorticotropin hormone-immunoreactive cells was significantly increased. General histological analysis of the adrenal cortex revealed a drastic thinning of the zona fasciculata and dystrophic changes in corticocytes. Structural changes, observed in the adrenal cortex, suggest a relative glucocorticoid deficiency, which may contribute to corticotropin-releasing hormone and adrenocorticotropin hormone upregulation in hypothalamus and pituitary of chronic heart failure patients. PMID- 19544246 TI - A randomized study to validate a midspinal canal depth nomogram in neonates. AB - Improving the accuracy of lumbar puncture (LP) in neonates should reduce the incidence of hemorrhagic contamination of cerebrospinal fluid (CSF) samples. We have previously demonstrated a linear correlation between neonatal weight and midspinal canal depth (MSCD), generating a nomogram and simple formula to allow MSCD estimation. In this study, we attempted to validate the nomogram by improving the quality of the CSF samples obtained. We consecutively randomized 99 infants in whom LP was clinically warranted to receive either a standard, "blind" ( N = 48) or "measured" ( N = 51) procedure. If allocated to the measured technique, the operator marked the LP needle with a Steri-Strip (TM) at the predicted depth of insertion (i.e., MSCD) derived from the weight-based nomogram. CSF samples were classified as clear (<500 red blood cells [rbc]/mL), mildly bloodstained (500 to 10,000 rbc/mL), heavily bloodstained (>10,000 rbc/mL or clotted), or failed procedures. Clear and mildly bloodstained LPs were "successful." Heavily bloodstained or failed procedures were considered "unsuccessful." We also recorded the number of attempts required to obtain a CSF sample. The overall success rate (or=4 positive nodes (P < 10(-5)). CONCLUSIONS: The pathological assessment of minimal 12 lymph nodes provides sufficient prognostic information. Three-tier nodal grouping is suggested for the next version of AJCC staging system for esophageal cancer. PMID- 19544365 TI - Enhanced fracture repair by leukotriene antagonism is characterized by increased chondrocyte proliferation and early bone formation: a novel role of the cysteinyl LT-1 receptor. AB - Inflammatory mediators and drugs which affect inflammation can influence the healing of injured tissues. Leukotrienes are potent inflammatory mediators, and similar to prostaglandins, are metabolites of arachidonic acid which can have positive or negative effects on bone and cartilage tissues. Here we tested the hypothesis that blocking the negative regulation of leukotrienes, would lead to enhanced endochondral bone formation during fracture repair. A closed femoral fracture was created in mice. Animals were divided into three groups for treatment with either montelukast sodium, a cysteinyl leukotriene type 1 receptor antagonist (trade name Singulair), zileuton, a 5-lipoxygenase enzyme inhibitor (trade name Zyflo), or carrier alone. The fractures were analyzed using radiographs, quantitative gene expression, histology and histomorphometry, and immunohistochemistry. Both the montelukast sodium group and the zileuton group exhibited enhanced fracture repair when compared with controls. Both treatment groups exhibited increased callous size and earlier bone formation when compared to controls as early as day 7. Gene expression analysis of treatment groups showed increased markers of chondrocyte proliferation and differentiation, and increased early bone formation markers when compared with controls. Treatment with montelukast sodium directly targeted the cysteinyl leukotriene type 1 receptor, leading to increased chondrocyte proliferation at early time points. These novel findings suggests a potential mechanism by which the cysteinyl leukotriene type 1 receptor acts as a negative regulator of chondrocyte proliferation, with important and previously unrecognized implications for both fracture repair, and in a broader context, systemic chondrocyte growth and differentiation. PMID- 19544366 TI - The effect of bulking agents on the chemical stability of acid-sensitive compounds in freeze-dried formulations: sucrose inversion study. AB - The goal of the study was to evaluate the impact of amorphous bulking agents on the chemical stability of freeze-dried materials. Polyvinylpyrrolidone and dextran of different molecular weights and lactose were used as bulking agents, and sucrose was used as an example of an acid-sensitive compound. Lyophiles containing bulking agent and sucrose at 10:1 (w/w) ratio, citrate buffer, and optionally bromophenol blue (pH indicator) were tested by X-ray powder diffractometry, differential scanning calorimetry, and Karl Fischer titrimetry. Diffuse reflectance UV-vis spectroscopy was used to obtain the concentration ratio of the deprotonated (In(2-)) to the protonated (HIn(-)) indicator species, from which the Hammett acidity function (H(2-)) was calculated. The extent of sucrose inversion in lyophiles stored at 60 degrees C was quantified by HPLC. The bulking agent had a major impact on both the apparent solid-state acidity (H(2-)) and the degradation rate, with the degradation rate constants value highest for dextran lyophiles (most "acidic", lower H(2-)) followed by lactose and polyvinylpyrrolidone lyophile (least "acidic", higher H(2-)). The Hammett acidity function can be used as an empirical solid-state acidity scale, to predict the rank-order stability of acid-sensitive compounds in lyophiles prepared with different bulking agents. PMID- 19544367 TI - Relative uptake of minoxidil into appendages and stratum corneum and permeation through human skin in vitro. AB - We examined uptake of the model therapeutic agent, minoxidil, into appendages, stratum corneum (SC), and through human skin, under the influence of different vehicles. Quantitative estimation of therapeutic drug deposition into all three areas has not previously been reported. Finite doses of minoxidil (2%, w/v) in formulations containing varying amounts of ethanol, propylene glycol (PG), and water (60:20:20, 80:20:0, and 0:80:20 by volume, respectively) were used. Minoxidil in SC (by tape stripping), appendages (by cyanoacrylate casting), and receptor fluid was determined by liquid scintillation counting. At early times (30 min, 2 h), ethanol-containing formulations (60:20:20 and 80:20:0) caused significantly greater minoxidil retention in SC and appendages, compared to the formulation lacking ethanol (0:80:20). A significant increase in minoxidil receptor penetration occurred with the PG-rich 0:80:20 formulation after 12 h. We showed that deposition of minoxidil into appendages, SC, and skin penetration into receptor fluid were similar in magnitude. Transport by the appendageal route is likely to be a key determinant of hair growth promotion by minoxidil. PMID- 19544368 TI - Paracetamol-loaded poly(epsilon-caprolactone) layered silicate nanocomposites prepared using hot-melt extrusion. AB - Composites of paracetamol loaded poly(epsilon-caprolactone) with layered silicates (nanoclays) were prepared using hot-melt extrusion. The paracetamol crystals and layered silicates formed both intercalated and partially exfoliated nanocomposite morphologies depending on composition. The dissolution and initial burst effect were retarded slightly by the nanoclay. T(m) and T(c) of poly(caprolactone) (PCL) were unaffected by the presence of nanoclay, but the crystalline content decreased. The highly dispersed nanoplatelets hindered the mobility of PCL chains and alter the crystallization behavior of PCL. The T(g) of PCL increased by up to 15 degrees C on addition of a synthetic fluromica, as the nanoclay constrained chain motion and tethered PCL chains through hydrogen bonding to hydroxyl groups on the edges of the clay platelets. The tensile mechanical properties of PCL were unaffected when a naturally derived clay (montmorillonite) and paracetamol were blended. In contrast, the modulus of PCL increased by 500% and the stress and elongation at break decreased by 30% for composites prepared with a partially synthetic fluoromica. The study has therefore demonstrated that nanocomposite formation is a potentially highly useful means of manipulating the mechanical properties of melt extrusion systems. PMID- 19544369 TI - Evidence of partial unfolding of proteins at the ice/freeze-concentrate interface by infrared microscopy. AB - The goal of this research was to use infrared spectroscopy in combination with a freeze drying stage to gain a better understanding of the mechanism of loss of protein integrity due to the stresses associated with freezing. Infrared spectra were collected in triplicate for the interstitial space between ice crystals and through ice crystals in a partially frozen system. Spectra were collected for lactate dehydrogenase (LDH) and human immune globulin (IgG) both in the presence and absence of an added surfactant (polysorbate 80). Spectra collected in the interstitial space, distant from the surface of ice crystals, were very similar to spectra collected from the initial solution regardless of the presence of a surfactant. Spectra collected through ice crystals, without added surfactant, were significantly different than spectra collected from the initial solution. An increase in bands characteristic of intermolecular beta-sheet structures (main component of aggregates) were present in these spectra. The presence of surfactant in both protein formulations resulted in a decrease in intermolecular beta-sheet signals in spectra of the proteins on the ice crystal surface. Additionally, much of the native state structure of LDH initially lost on the surface of ice crystals returned when surfactant was added to the formulation prior to freezing. PMID- 19544370 TI - Imaging pharmaceutical tablets with optical coherence tomography. AB - Optical coherence tomography (OCT) is a recently developed optical technique that produces depth profiles of three-dimensional objects. It is a nondestructive interferometric method responding to refractive index variation in the sample under study and can reach a penetration depth of a few millimetres. OCT employs near-infrared (NIR) light and therefore provides a link between NIR spectroscopy and Terahertz (THz) measurements that are often used to characterise tablets. In this article we assess the potential of OCT as a reliable and practical tool in the analysis of pharmaceutical tablets and coatings. A variety of tablets were tested with different shapes, formulations and coatings. We consider the origins of contrast in the obtained images and demonstrate that it correlates strongly with the expected tablet structure. The influence of absorption and scattering are considered for the wavelength ranges used. The results show that OCT is a promising diagnostic tool with an important role to play in the tablet and coating technologies. The high measurement speed of OCT and its relative ease of implementation make it also an attractive candidate technology for in-line quality control during manufacturing. PMID- 19544371 TI - Vibration effects of lab equipment on dissolution testing with USP paddle method. AB - Environmental vibration induced by laboratory equipment, building construction, or even by the analysts themselves is one of the more complicated factors affecting dissolution testing. It is difficult to control and/or calibrate by mechanical means or performance-based methods. In this study, dissolution apparatus vibration levels were measured in the frequency range from 10 to 270 Hz along all three axes using commercially available, single-axis accelerometers. The vibration distribution on the dissolution vessel plate was mapped, and acceleration was subsequently measured during dissolution runs involving NCDA#2 (10 mg prednisone) tablets using the paddle method. Several types of laboratory equipment were used to induce vibration during dissolution testing and vibration levels along the X-, Y-, and Z-axes of the vessel plate were measured in an attempt to establish possible correlation with dissolution results. In the frequency range studied, root mean square (RMS) acceleration values above 0.01 g, in either vertical or horizontal direction, typically affected dissolution results. PMID- 19544373 TI - Vital microscopic analysis of polymeric micelle extravasation from tumor vessels: macromolecular delivery according to tumor vascular growth stage. AB - Particles larger than a specific size have been thought to extravasate from tumor vessels but not from normal vessels. Therefore, various nanoparticles incorporating anticancer drugs have been developed to realize selective drug delivery to solid tumors. However, it is not yet clear whether nanoparticles extravasate readily from all tumor vessels including vessels of microtumors. To answer this question, we synthesized new polymeric micelles labeled with fluorescein isothiocyanate (FITC) and injected them into the tail vein of rats with implanted skinfold transparent chambers. We also analyzed, by means of time lapse vital microscopy with image analysis, extravasation of FITC micelles from tumor vessels at different stages of growth of Yoshida ascites sarcoma LY80. Polymeric micelles readily leaked from vessels at the interface between normal and tumor tissues and those at the interface between tumor tissues and necrotic areas. The micelles showed negligible extravasation, however, from the vascular network of microtumors less than 1 mm in diameter and did not accumulate in the microtumor. Our results suggest that we must develop a novel therapeutic strategy that can deliver sufficient nanomedicine to microtumors. PMID- 19544372 TI - Magnetic resonance imaging-guided adoptive cellular immunotherapy of central nervous system tumors with a T1 contrast agent. AB - Dendritic cells (DCs) are the most effective antigen-presenting cells (APCs) and are used in a variety of immunotherapeutic approaches. Adoptive cellular immunotherapy (ACI) of cancer using DCs has attracted much interest due to their capacity to promote immunity in prophylactic and therapeutic protocols. As one approach, DCs are injected into patients or tumor-bearing animals, to trigger specific antitumor immunity. In that framework, several approaches to DC delivery have been reported, including direct intratumoral injection; this has yielded positive but variable results. The underlying reasons for this have not been fully determined, but major hypotheses include technical difficulties in delivering cells into tumors and tumor-mediated immunosuppression. Image-guided ACI offers the potential to establish that DCs are efficiently delivered to the tumor site, which might eliminate some of the variability. Therefore, we developed highly sensitive methods for monitoring the injection or trafficking of DCs into tumors using a clinically approved formulation of a gadolinium-based magnetic resonance imaging (MRI) contrast agent, Gd(III)-HP-DO3A (ProHance). We determined the labeling efficiency of DCs with this formulation; that labeling DCs with this agent did not inhibit expression of surface markers important for antigen presentation and activation of naive T cells; that their capacity to interact with natural killer (NK) cells was not reduced; and that their migration was not diminished. Further, we determined that ProHance-labeled DCs can be effectively imaged in vivo in established central nervous system tumors. PMID- 19544375 TI - Tailoring treatment strategy for acute promyelocytic leukemia in low-income countries. PMID- 19544374 TI - Modulation of the activity of ABC transporters (P-glycoprotein, MRP2, BCRP) by flavonoids and drug response. AB - The present article aims to review the up-to-date information on the most recent studies of the interaction of flavonoids with ABC transporters, in particular the drug pharmacokinetic consequences of such a relationship. In addition, the modulation of the expression of the ABC transporters by flavonoids is also illustrated. Flavonoids are a large group of plant polyphenols present extensively in our daily diets and herbal products. High intake of isoflavones has been associated with a variety of beneficial effects on several common diseases. These polyphenols interact with ABC drug transporters involved in drug resistance and drug absorption, distribution and excretion. A number of studies have demonstrated inhibition of drug transporters by flavonoids. This flavonoid ABC-transporter interaction could be beneficial for poorly absorbed drugs but could also result in severe drug intoxication, especially drugs with a narrow therapeutic window. On the other hand, flavonoids are themselves substrates of ABC transporters. These proteins can affect the oral availability and tissue distribution of these compounds, modifying their beneficial effects. The challenge is to find a suitable way to predict harmful drug-flavonoid interactions mediated by these transporters. PMID- 19544376 TI - More than physical function: HRQOL among children and adolescents with lower extremity bone sarcoma. PMID- 19544377 TI - Guidelines for Epo use in children with cancer. PMID- 19544378 TI - Revisiting the role of positron-emission tomography/computed tomography in determining the need for planned neck dissection following chemoradiation for advanced head and neck cancer. AB - OBJECTIVES/HYPOTHESIS: Planned neck dissection following chemoradiation (CR) has been advocated in patients with head and neck squamous cell cancer (HNSCC) with advanced nodal disease and a clinical complete response to CR because of the potential for residual occult nodal disease. The utility of positron-emission tomography/computed tomography (PET-CT) in identifying occult nodal disease in this scenario is controversial. METHODS: The medical records of all patients treated with CR for advanced HNSCC with N2 or N3 disease from December 2003 to June 2007 were reviewed. Patients with a complete clinical response were included if PET-CT performed 8 to 11 weeks after CR showed no distant disease and they underwent planned neck dissection. RESULTS: Thirty-two patients met study criteria. PET-CT was positive for residual nodal disease in 20 patients (63%). Pathology revealed carcinoma in 10 patients (31%): six of 20 patients with positive PET-CT scans (30%) and four of 12 patients with negative PET-CT scans (33%). The sensitivity and specificity of PET-CT was 60% and 36%. Regional recurrence developed in two patients (6%) who were not successfully salvaged. CONCLUSIONS: PET-CT performed 8 to 11 weeks after CR does not reliably predict the need for planned post-treatment neck dissection in patients with a complete clinical response following CR. Regional recurrence rates are comparable to those reported for patients observed with PET-CT, suggesting no advantage for planned neck dissection, and salvage rates were poor. These data suggest that delaying the timing of PET-CT, with surgery reserved for positive findings, is a reasonable alternative to planned neck dissection to avoid unnecessary surgery. PMID- 19544379 TI - The assessment of pharyngocutaneous fistula rate in patients treated primarily with definitive radiotherapy followed by salvage surgery of the larynx and hypopharynx. AB - OBJECTIVES/HYPOTHESIS: To determine whether definitive radiotherapy prior to surgery increases the rate of pharyngocutaneous fistula (PCF) following laryngectomy or hypopharyngectomy and to determine if differences in duration of time between definitive radiotherapy and surgery alters PCF rate. STUDY DESIGN: A retrospective review of 152 patients treated surgically for primary laryngeal or hypopharyngeal squamous cell carcinoma. METHODS: Following previous definitive radiotherapy treatment 38 patients underwent salvage surgery and 114 patients underwent primary surgery with curative intent. The PCF rate was assessed in both groups. RESULTS: The rate of PCF was found to be significantly higher in the salvage surgery group than those undergoing primary surgery (34.2% vs. 15.7%) (P < .05). Fistula rate was also higher in the subgroup that received concurrent chemoradiation to radiotherapy alone (P = .002). The patients who developed PCF in the salvage surgery group had significantly lower median time to surgery (5.8 months) than the nonfistula group (9.8 months) (P = .032). PCF rate was 75% within 4 months of radiotherapy to salvage surgery compared to 25% after 4 months (P = .034). Within 12 months of radiotherapy this percentage was 48% compared to 0% after 12 months (P = .014). The median radiotherapy dose was significantly higher in those whose surgery was complicated by PCF (70 Gy) compared to patients who did not develop a fistula (64 Gy) (P = .001). CONCLUSIONS: Patients undergoing salvage surgery within 12 months, and in particular within 4 months, who have received high dose radiotherapy (>64 Gy) or concurrent chemoradiation are at high risk of developing PCF. PMID- 19544381 TI - Characterization and gene expression profiling in glioma cell lines with deletion of chromosome 19 before and after microcell-mediated restoration of normal human chromosome 19. AB - Nearly 10% of human gliomas are oligodendrogliomas. Deletion of chromosome arm 19q, often in conjunction with deletion of 1p, has been observed in 65-80% of these tumors. This has suggested the presence of a tumor suppressor gene located on the 19q arm. Chromosome 19 deletion is also of interest due to the better prognosis of patients with deletion, including longer survival and better response to chemotherapy, compared with patients without deletion. Two glioma cell lines with deletion of 19q were used for chromosome 19 microcell-mediated transfer, to assess the effect of replacing the deleted segment. Complementation with chromosome 19 significantly reduced the growth rate of the hybrid cells compared with the parental cell lines. Affymetrix U133 Plus 2.0 Gene Chip analysis was performed to measure and compare the expression of the chromosome 19 genes in the chromosome 19 hybrid cell lines to the parental cell line. Probes were considered significantly different when a P value <0.01 was seen in all of the cell line comparisons. Of 345 probes within the commonly deleted 19q region, seven genes (APOE, RCN3, FLJ10781, SAE1, STRN4, CCDC8, and BCL2L12) were identified as potential candidate genes. RT-PCR analysis of primary tumor specimens showed that several genes had significant differences when stratified by tumor morphology or deletion status. This suggests that one or more of these candidates may play a role in glioma formation or progression. PMID- 19544380 TI - A controlled safety study of diindolylmethane in the immature rat model. AB - OBJECTIVES/HYPOTHESIS: Diindolylmethane (DIM), a natural product from cruciferous vegetables, has been shown to be a dietary component that has inhibitory effects on some tumors (e.g., laryngeal papilloma). However, current evidence to support its safety is based on adult humans or mature animals. There is little to show on its safety in children. This study is designed to assess safety in the young rat model. STUDY DESIGN: Prospective controlled animal study. METHODS: Forty rats were separated into four treatment groups of 10 rats each, based on the amount of study drug they received in their daily food: 1) immature rats fed a low dose of DIM, (0.6 mg/kg/day); 2) immature rats fed a high dose of DIM (6.0 mg/kg/day); 3) immature rats fed no DIM (control); and 4) adult rats fed a high dose of DIM (6.0 mg/kg/day). At the conclusion of the study we collected blood to compare serum chemistries and vitamin D levels, and harvested organs to observe for any gross or histological changes among the groups. Statistical methods involved one-way analysis of variance and pairwise comparisons with Tukey multiple comparison adjustment. RESULTS: Although our numbers do not allow for statistical significance, there was no appreciable difference in rat weights among the immature groups, nor was there appreciable difference in serum chemistries, or gross or histological examination of liver, kidney, and bone. CONCLUSIONS: Diindolylmethane seems to have no adverse affects on the rat even when given in doses 3x what we propose to be therapeutic. This adds evidence to the safety of this drug in the pediatric population as a treatment option for recurrent respiratory papilloma. PMID- 19544382 TI - Involvement of cathepsin B in the processing and secretion of interleukin-1beta in chromogranin A-stimulated microglia. AB - Cathepsin B (CB) is a cysteine lysosomal protease implicated in a number of inflammatory diseases. Although it is now evident that caspase-1, an essential enzyme for maturation of interleukin-1beta (IL-1beta), can be activated through the inflammasome, there is still evidence suggesting the existence of lysosomal proinflammatory caspase pathways. In the present study, a marked induction of pro IL-1beta, its processing to the mature form and secretion were observed in the primary cultured microglia prepared from wild-type mice after stimulation with chromogranin A (CGA). Although pro-IL-1beta also markedly increased in microglia prepared from CB-deficient mice, CB-deficiency abrogated the pro-IL-1beta processing. CA-074Me, a specific inhibitor for CB, inhibited the pro-IL-1beta maturation and its release from microglia. Furthermore, the caspase-1 activation was also inhibited by CA-074Me and E-64d, a broad cysteine protease inhibitor. After treatment with CGA, CB was markedly induced at both protein and mRNA levels. The induced pro-CB was rapidly processed to its mature form. The immunoreactivity for CB co-localized with both that for caspase-1 and the cleaved IL-1beta, in the acidic enlarged lysosomes. Inconsistent with these in vitro observations, the immunoreactivity for the cleaved IL-1beta was markedly observed in microglia of the hippocampus from aged wild-type but not CB-deficient mice. These observations strongly suggest that CB plays a key role in the pro-IL-1beta maturation through the caspase-1 activation in enlarged lysosomes of CGA-treated microglia. Therefore, either pharmacological or genetic inhibition of CB may provide therapeutic intervention in inflammation-associated neurological diseases. PMID- 19544384 TI - Hydrogenotrophic denitrification and perchlorate reduction in ion exchange brines using membrane biofilm reactors. AB - Halophilic (salt loving), hydrogenotrophic (H(2) oxidizing) denitrifying bacteria were investigated for treatment of nitrate (NO3-) and perchlorate (ClO4-) contaminated groundwater and ion exchange (IX) brines. Hydrogenotrophic denitrifying bacteria were enriched from a denitrifying wastewater seed under both halophilc and non-halophilc conditions. The cultures were inoculated into bench-scale membrane biofilm reactors (MBfRs) with an "outside in" configuration, with contaminated water supplied to the lumen of the membranes and H(2) supplied to the shell. Abiotic mass transfer tests showed that H(2) mass transfer coefficients were lower in brines than in tap water at highest Reynolds number, possibly due to increased transport of salts and decreased H(2) solubility at the membrane/liquid interface. An average NO3- removal efficiency of 93% was observed for the MBfR operated in continuous flow mode with synthetic contaminated groundwater. Removal efficiencies of 30% for NO3- and 42% for ClO4- were observed for the MBfR operated with synthetic IX brine in batch operating mode with a reaction time of 53 h. Phylogenetic analysis focused on the active microbial community and revealed that halotolerant, NO3- -reducing bacteria of the bacterial classes Gamma-Proteobacteria and Sphingobacteria were the metabolically dominant members within the stabilized biofilm. This study shows that, despite decreased H(2) transfer under high salt conditions, hydrogenotrophic biological reduction may be successfully used for the treatment of NO3- and ClO- in a MBfR. PMID- 19544385 TI - Reduced L-arginine transport contributes to the pathogenesis of myocardial ischemia-reperfusion injury. AB - Myocardial injury due to ischemia-reperfusion (I-R) damage remains a major clinical challenge. Its pathogenesis is complex including endothelial dysfunction and heightened oxidative stress although the key driving mechanism remains uncertain. In this study we tested the hypothesis that the I-R process induces a state of insufficient L-arginine availability for NO biosynthesis, and that this is pivotal in the development of myocardial I-R damage. In neonatal rat ventricular cardiomyocytes (NVCM), hypoxia-reoxygenation significantly decreased L-arginine uptake and NO production (42 +/- 2% and 71 +/- 4%, respectively, both P < 0.01), maximal after 2 h reoxygenation. In parallel, mitochondrial membrane potential significantly decreased and ROS production increased (both P < 0.01). NVCMs infected with adenovirus expressing the L-arginine transporter, CAT1, and NVCMs supplemented with L-arginine both exhibited significant (all P < 0.05) improvements in NO generation and mitochondrial membrane potentials, with a concomitant significant fall in ROS production and lactate dehydrogenase release during hypoxia-reoxygenation. In contrast, L-arginine deprived NVCM had significantly worsened responses to hypoxia-reoxygenation. In isolated perfused mouse hearts, L-arginine infusion during reperfusion significantly improved left ventricular function after I-R. These improved contractile responses were not dependent on coronary flow but were associated with a significant decrease in nitrotyrosine formation and increases in phosphorylation of both Akt and troponin I. Collectively, these data strongly implicate reduced L-arginine availability as a key factor in the pathogenesis of I-R injury. Increasing L-arginine availability via increased CAT1 expression or by supplementation improves myocardial responses to I-R. Restoration of L-arginine availability may therefore be a valuable strategy to ameliorate I-R injury. PMID- 19544383 TI - Fungal rhinosinusitis: a categorization and definitional schema addressing current controversies. AB - BACKGROUND: Fungal (rhino-) sinusitis encompasses a wide spectrum of immune and pathological responses, including invasive, chronic, granulomatous, and allergic disease. However, consensus on terminology, pathogenesis, and optimal management is lacking. The International Society for Human and Animal Mycology convened a working group to attempt consensus on terminology and disease classification. DISCUSSION: Key conclusions reached were: rhinosinusitis is preferred to sinusitis; acute invasive fungal rhinosinusitis is preferred to fulminant, or necrotizing and should refer to disease of <4 weeks duration in immunocompromised patients; both chronic invasive rhinosinusitis and granulomatous rhinosinusitis were useful terms encompassing locally invasive disease over at least 3 months duration, with differing pathology and clinical settings; fungal ball of the sinus is preferred to either mycetoma or aspergilloma of the sinuses; localized fungal colonization of nasal or paranasal mucosa should be introduced to refer to localized infection visualized endoscopically; eosinophilic mucin is preferred to allergic mucin; and allergic fungal rhinosinusitis (AFRS), eosinophilic fungal rhinosinusitis, and eosinophilic mucin rhinosinusitis (EMRS) are imprecise and require better definition. In particular, to implicate fungi (as in AFRS and EMRS), hyphae must be visualized in eosinophilic mucin, but this is often not processed or examined carefully enough by histologists, reducing the universality of the disease classification. A schema for subclassifying these entities, including aspirin-exacerbated rhinosinusitis, is proposed allowing an overlap in histopathological features, and with granulomatous, chronic invasive, and other forms of rhinosinusitis. Recommendations for future research avenues were also identified. PMID- 19544386 TI - Activation and deactivation of periventricular white matter phagocytes during postnatal mouse development. AB - Brain microglia are related to peripheral macrophages but undergo a highly specific process of regional maturation and differentiation inside the brain. Here, we examined this deactivation and morphological differentiation in cerebral cortex and periventricular subcortical white matter, the main "fountain of microglia" site, during postnatal mouse development, 0-28 days after birth (P0 P28). Only macrophages in subcortical white matter but not cortical microglia exhibited strong expression of typical activation markers alpha5, alpha6, alphaM, alphaX, and beta2 integrin subunits and B7.2 at any postnatal time point studied. White matter phagocyte activation was maximal at P0, decreased linearly over P3 and P7 and disappeared at P10. P7 white matter phagocytes also expressed high levels of IGF1 and MCSF, but not TNFalpha mRNA; this expression disappeared at P14. This process of deactivation followed the presence of ingested phagocytic material but correlated only moderately with ramification, and not with the extent of TUNEL+ death in neighboring cells, their ingestion or microglial proliferation. Intravenous fluosphere labeling revealed postnatal recruitment and transformation of circulating leukocytes into meningeal and perivascular macrophages as well as into ramified cortical microglia, but bypassing the white matter areas. In conclusion, this study describes strong and selective activation of postnatally resident phagocytes in the P0-P7 subcortical white matter, roughly equivalent to mid 3rd trimester human fetal development. This presence of highly active and IGF1- and MCSF-expressing phagocytes in the neighborhood of vulnerable white matter could play an important role in the genesis of or protection against axonal damage in the fetus and premature neonate. PMID- 19544387 TI - Varied opinions on thrombolysis for venous thromboembolism in infants and children: findings from a survey of pediatric hematology-oncology specialists. AB - BACKGROUND: Recent guidelines discourage routine use of thrombolytic agents for treatment of venous thromboembolism (VTE) in pediatric patients, but actual practice patterns are unknown. PROCEDURE: An electronic survey was emailed to all active and trainee members of the American Society of Pediatric Hematology/Oncology in April 2008. Respondents were asked a series of multiple choice questions based on hypothetical case scenarios describing pediatric VTE, pertinent to the implementation of thrombolytic therapy and other professional demographic information. RESULTS: Two hundred eighty-five evaluable responses were obtained (22% response rate) which varied greatly with respect to all spheres of questioning. Tissue plasminogen activator (tPA) was the thrombolytic agent chosen by most respondents, but no clear consensus emerged as to appropriate indications (although preference for thrombolytic therapy increased with severity of the posed clinical scenario), mode of tPA delivery (systemic vs. catheter-directed), dose (high-dose vs. low-dose regimen) or a suitable maximum duration of therapy (range: 1-168 hr; varied according to specific dosing regimen chosen). Expertise in pediatric thrombosis, years out from fellowship training and volume of experience with cases of pediatric thrombosis were not largely associated with respondent choices; however, institutional experience with pharmacologic thrombolysis exhibited the most notable association of the professional demographic factors analyzed. CONCLUSIONS: The survey results support that clinical practice pertaining to use of thrombolytic agents in pediatric VTE varies widely but also provide useful benchmarks to aid clinical decision-making and future clinical trial design. Such varied practices stem from the lack of strong evidence supporting one therapeutic approach versus another. PMID- 19544389 TI - Validation of a new method for determination of cup orientation in THA. AB - Our goal was to validate accuracy, consistency, and reproducibility/reliability of a new method for determining cup orientation in total hip arthroplasty (THA). This method allows matching the 3D-model from CT images or slices with the projected pelvis on an anteroposterior pelvic radiograph using a fully automated registration procedure. Cup orientation (inclination and anteversion) is calculated relative to the anterior pelvic plane, corrected for individual malposition of the pelvis during radiograph acquisition. Measurements on blinded and randomized radiographs of 80 cadaver and 327 patient hips were investigated. The method showed a mean accuracy of 0.7 +/- 1.7 degrees (-3.7 degrees to 4.0 degrees) for inclination and 1.2 +/- 2.4 degrees (-5.3 degrees to 5.6 degrees) for anteversion in the cadaver trials and 1.7 +/- 1.7 degrees (-4.6 degrees to 5.5 degrees) for inclination and 0.9 +/- 2.8 degrees (-5.2 degrees to 5.7 degrees) for anteversion in the clinical data when compared to CT-based measurements. No systematic errors in accuracy were detected with the Bland Altman analysis. The software consistency and the reproducibility/reliability were very good. This software is an accurate, consistent, reliable, and reproducible method to measure cup orientation in THA using a sophisticated 2D/3D matching technique. Its robust and accurate matching algorithm can be expanded to statistical models. PMID- 19544388 TI - Sustained epidermal growth factor receptor levels and activation by tethered ligand binding enhances osteogenic differentiation of multi-potent marrow stromal cells. AB - Epidermal growth factor receptor (EGFR)-mediated signaling helps regulate bone development and healing through its effects on osteogenic cells. Here, we show how EGFR activity and osteogenic differentiation responses in primary human bone marrow-derived multipotent stromal cells (MSCs) are influenced by presenting covalently tethered epidermal growth factor (tEGF) on the culture substratum, a presentation mode that reduces EGFR internalization and restricts signaling to the cell surface. In both absence and presence of tEGF, MSCs increase expression levels of EGFR and its heterodimerization partner HER2 during the course of osteogenic differentiation. tEGF substrata increased levels of phosphorylated EGFR and phosphorylated extracellular regulated kinase (ERK) compared to control substrata, and these elevations were associated with a twofold enhancement of MSC alkaline phosphatase activity at day 7 and matrix mineralization at day 21. Surprisingly, addition of soluble EGF (sEGF) to cells cultured on tEGF substrata reduces osteogenic differentiation, even though EGFR signaling is more strongly activated in acute, short-term manner by sEGF treatment than by tEGF treatment. A striking concomitant result of the sEGF effects is near-complete downregulation of EGFR and HER2, demonstrating that the tEGF/EGFR interaction is dynamically reversible even though temporally sustained. Taken together, our results show that enhanced MSC osteogenic differentiation corresponds to a sustained combination of receptor expression and ligand presentation, both of which are maintained by tEGF. PMID- 19544390 TI - Growth patterns in children with sickle cell anemia during puberty. AB - BACKGROUND: Previous studies of children with homozygous sickle cell anemia (SCA) show impaired growth and maturation. The correlation of this suboptimal growth with metabolic and hematological factors during puberty is poorly understood. PROCEDURE: We studied a group of pre-adolescent children with SCA (19 males, 14 females) and healthy controls (16 males, 15 females) matched for race, sex, body size, and pubertal development. Height, weight, body mass index (BMI), and body composition changes were longitudinally assessed over a 2-year period and compared between the groups and with Z scores based on US growth charts. These changes were correlated with hemoglobin (Hgb) concentration and with energy expenditure (EE) measured using indirect whole-room calorimetry. RESULTS: Children with SCA progressed through puberty slower than control children. While, after 2 years, pubertal males with SCA were shorter, their annual increases in weight were not different from controls. The mean fat free mass (FFM) increments were significantly less in males and females with SCA than in control children. In males with SCA, growth in height declined over time and was significantly slower than in matched controls (P < 0.05). CONCLUSION: Growth delays were present during puberty in children with SCA. Decreased growth velocity in children with SCA was independently associated with decreased Hgb concentration and increased total EE. PMID- 19544391 TI - Thrombotic thrombocytopenic purpura in childhood. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare disease, especially in childhood, and has a high mortality rate in the absence of appropriate treatment. It is characterised by microangiopathic haemolytic anaemia and consumptive thrombocytopenia. TTP may be difficult to distinguish from haemolytic uraemic syndrome (HUS) because of similar clinical manifestations and laboratory findings. In the past, TTP and HUS have often been considered to represent variable expressions of a single entity. Our increased understanding of the pathogenesis of TTP has in turn resulted in significant improvements in its treatment and outcomes. Several immunomodulating agents are currently being used with variable outcomes. PMID- 19544392 TI - Hydrogen sulfide-releasing NSAIDs attenuate neuroinflammation induced by microglial and astrocytic activation. AB - Endogenously generated hydrogen sulfide (H(2)S) may have multiple functions in brain. It has been shown that H(2)S attenuates the expression of pro-inflammatory cytokines by lipopolysaccharide (LPS)-activated microglia. Here we demonstrate a neuroprotective effect of NaSH and three H(2)S-releasing compounds, ADT-OH, S diclofenac, and S-aspirin. When activated by LPS and gamma-interferon, human microglia and THP-1 cells release materials that are toxic to human neuroblastoma SH-SY5Y cells. These phenomena also occur with gamma-interferon-stimulated human astroglia and U118 cells. When these cell types are pretreated with aspirin, diclofenac, NASH, or ADT-OH, the supernatants are significantly less toxic. When they are treated with the NSAID-H(2)S hybrid molecules S-diclofenac and S aspirin, which are here referred to as S-NSAIDs, there is a significant enhancement of the protection. The effect is concentration and incubation time dependent. Such pretreatment also reduces the release of the proinflammatory mediators TNFalpha, IL-6, and nitric oxide. The H(2)S-releasing compounds are without effect when applied directly to SH-SY5Y cells. These data suggest that hybrid H(2)S releasing compounds have significant antiinflammatory properties and may be candidates for treating neurodegenerative disorders that have a prominent neuroinflammatory component such as Alzheimer disease and Parkinson disease. PMID- 19544393 TI - Humoral immunity to diphtheria, tetanus, measles, and hemophilus influenzae type b in children with acute lymphoblastic leukemia and response to re-vaccination. AB - OBJECTIVE: Loss of immunity to previous vaccination and timing of re-vaccination in children receiving chemotherapy remains controversial. The aim of this study was to investigate the immunity to vaccine preventable diseases in children with acute lymphoblastic leukemia (ALL). PROCEDURE: Sixty-one patients with ALL and 13 healthy siblings were enrolled. Three study groups included newly diagnosed patients (group 1), patients on maintenance chemotherapy (group 2), and patients that completed chemotherapy (group 3). Blood samples for baseline antibody titers were obtained from all the patients and controls. Patients in group 2 were vaccinated with diphtheria, tetanus, and hemophilus influenzae type b (Hib). Patients in group 3 and controls received the measles vaccine in addition to all the above vaccines. In groups 2 and 3, post-vaccination antibody titers were also obtained. RESULTS: Patients and controls had no Hib vaccine during primary vaccination. After chemotherapy median antibody levels against diphtheria, tetanus, measles, and Hib were decreased but tetanus antibodies were still at the protective levels. Proportions of the patients with protective levels were 11.1%, 83.3%, 16.7%, and 16.7% for diphtheria, tetanus, Hib, and measles, respectively. Vaccination achieved protective antibody levels in 81%, 100%, 89.5%, and 70% of the patients for diphtheria, tetanus, Hib, and measles, respectively. Vaccine responses during maintenance were also satisfying. CONCLUSION: We recommend re vaccination after 3 months of cessation of chemotherapy. Administration of Hib vaccine may be beneficial after the first 3 months of maintenance chemotherapy especially in children with no primary vaccination followed by a second booster dose after cessation of therapy to increase immunity. PMID- 19544394 TI - Expression of antigen processing and presenting molecules by Schwann cells in inflammatory neuropathies. AB - Schwann cells are the myelinating glia cells of the peripheral nervous system (PNS) and can become targets of an autoimmune response in inflammatory neuropathies like the Guillain-Barre syndrome (GBS). Professional antigen presenting cells (APCs) are known to promote autoimmune responses in target tissues by presenting self-antigens. Other cell types could participate in local autoimmune responses by acting as nonprofessional APCs. Using a combined approach of immunocytochemistry, immunohistochemistry, and flow cytometry analysis we demonstrate that human Schwann cells express the antigen processing and presenting machinery (APM) in vitro and in vivo. Moreover, cultured human Schwann cells increase the expression of proteasome subunit delta (Y), antigen peptide transporter TAP2, and HLA Class I and HLA Class II complexes in an inflammatory environment. In correlation with this observation, Schwann cells in sural nerve biopsies from GBS patients show increased expression of antigen processing and presenting molecules. Furthermore, cultured human Schwann cells can proteolytically digest fluorescently-labeled nonmammalian antigen ovalbumin. Taken together, our data suggest antigen processing and presentation as a possible function of Schwann cells that may contribute to (auto)immune responses within peripheral nerves. PMID- 19544396 TI - Imaging and spectroscopic findings in meningioangiomatosis. AB - Meningioangiomatosis (MA) is an uncommon brain tumor. The role of imaging techniques is underscored in cases where the tumor location makes resection (or even biopsy) dangerous. We report the case of a child with an MA tumor located deep in the right sylvian fissure. A computed tomography (CT) scan showed calcifications in a highly vascular lesion with surrounding edema. Magnetic resonance spectroscopy (MRS) showed a distinct choline (Cho) peak, which usually suggests a proliferating tumor. Fluorodeoxyglucose positron emission tomography (FDG-PET) showed the lesion lacked hypermetabolic features. These radiological features should put MA in the differential diagnosis. PMID- 19544397 TI - Evaluation of adipose-derived stromal vascular fraction or bone marrow-derived mesenchymal stem cells for treatment of osteoarthritis. AB - The purpose of this study was the assessment of clinical, biochemical, and histologic effects of intraarticular administered adipose-derived stromal vascular fraction or bone marrow-derived mesenchymal stem cells for treatment of osteoarthritis. Osteoarthritis was induced arthroscopically in the middle carpal joint of all horses, the contralateral joint being sham-operated. All horses received treatment on Day 14. Eight horses received placebo treatment and eight horses received adipose-derived stromal vascular fraction in their osteoarthritis affected joint. The final eight horses were treated the in osteoarthritis affected joint with bone marrow-derived mesenchymal stem cells. Evaluations included clinical, radiographic, synovial fluid analysis, gross, histologic, histochemical, and biochemical evaluations. No adverse treatment-related events were observed. The model induced a significant change in all but two parameters, no significant treatment effects were demonstrated, with the exception of improvement in synovial fluid effusion PGE2 levels with bone marrow-derived mesenchymal stem cells when compared to placebo. A greater improvement was seen with bone marrow-derived mesenchymal stem cells when compared to adipose-derived stromal vascular fraction and placebo treatment. Overall, the findings of this study were not significant enough to recommend the use of stem cells for the treatment of osteoarthritis represented in this model. PMID- 19544395 TI - Maintaining retinal astrocytes normalizes revascularization and prevents vascular pathology associated with oxygen-induced retinopathy. AB - Astrocytes are well known modulators of normal developmental retinal vascularization. However, relatively little is known about the role of glial cells during pathological retinal neovascularization (NV), a leading contributor to vision loss in industrialized nations. We demonstrate that the loss of astrocytes and microglia directly correlates with the development of pathological NV in a mouse model of oxygen-induced retinopathy (OIR). These two distinct glial cell populations were found to have cooperative survival effects in vitro and in vivo. The intravitreal injection of myeloid progenitor cells, astrocytes, or astrocyte-conditioned media rescued endogenous astrocytes from degeneration that normally occurs within the hypoxic, vaso-obliterated retina following return to normoxia. Protection of the retinal astrocytes and microglia was directly correlated with accelerated revascularization of the normal retinal plexuses and reduction of pathological intravitreal NV normally associated with OIR. Using astrocyte-conditioned media, several factors were identified that may contribute to the observed astrocytic protection and subsequent normalization of the retinal vasculature, including vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Injection of VEGF or bFGF at specific doses rescued the retinas from developing OIR-associated pathology, an effect that was also preceded by protection of endogenous glia from hypoxia-induced degeneration. Together, these data suggest that vascular-associated glia are also required for normalized revascularization of the hypoxic retina. Methods developed to target and protect glial cells may provide a novel strategy by which normalized revascularization can be promoted and the consequences of abnormal NV in retinal vascular diseases can be prevented. PMID- 19544399 TI - Differentiation of synovial CD-105(+) human mesenchymal stem cells into chondrocyte-like cells through spheroid formation. AB - Mesenchymal stem cells (MSCs) have the capacity to differentiate into several cell lineages, some of which can generate bone, cartilage, or adipose tissue. The presence of MSCs in the synovial membrane was recently reported. Data from comparative studies of MSCs derived from various mesenchymal tissues suggest that MSCs from synovial membranes have a superior chondrogenesis capacity. Previous chondrogenic differentiation studies have used the total population of MSCs, including cells with several MSC markers, such as CD44, CD90, CD105, or CD73. However the chondrogenic capacity of an individual population of MSCs has not been examined. Our aim was to study the chondrogenic capacity of the cellular MSC subset, CD105(+), derived from synovial membrane tissues of patients with osteoarthritis (OA) and normal donors. The tissues were digested with a cocktail of collagenase/dispase and the isolated MSCs were seeded into plates. The subpopulation of CD105(+)-MSCs was separated using a magnetic separator. The MSCs were then differentiated towards chondrocyte-like cells using a specific medium to promote spheroid formation. Spheroids were collected after 14, 28, and 46 days in chondrogenic medium and stained with hematoxylin, eosin, Safranin O or Alcian blue to evaluate the extracellular matrix. Immunohistochemistry was performed to study collagen types I (COLI) and II (COLII) and aggrecan expression. Phenotypic characterization of the isolated CD105(+)-MSCs shows that these cells are also positive for CD90 and CD44, but negatives for CD34 and CD45. In addition, this cellular subset expressed Sox-9. Spheroids appeared after 7 days in culture in the presence of chondrogenic medium. Our studies show no differences between MSCs obtained from OA and normal synovial membranes during chondrogenesis. The morphological analysis of spheroids revealed characteristics typical of chondrocyte cells. The intensity of Safranin O, Alcian blue and aggrecan staining was positive and constant throughout the culture period. However, the intensity of COL2 staining was higher at 28 days (84.29 +/- 0.1 U) than at 46 days (61.28 +/- 01 U), while COL1 staining was not detected in any samples analyzed. These results were confirmed by reverse transcriptase-polymerase chain reaction assays. We conclude that the cellular subset of CD105(+)-MSCs has chondrogenic capacity. The study also show the similar chondrogenic capacity of CD105(+)-MSCs cultured from normal and OA synovial membranes. PMID- 19544398 TI - Phosphoproteome sequence analysis and significance: mining association patterns around phosphorylation sites utilizing MAPRes. AB - Phosphorylation, one of the most common protein post-translational modifications (PTMs) on hydroxyl groups of S/T/Y is catalyzed by kinases and involves the presence or absence of certain amino acid residues in the vicinity of the phosphorylation sites. Using MAPRes, we have analyzed the substrate proteins of Phospho.ELM 7.0 and found that there are both general and specific requirements for the presence or absence of particular amino acids in the vicinity of phosphorylated S/T/Y for both of the phosphorylation data, whether or not kinase information was taken into account. Patterns extracted by MAPRes for kinase specific data have been utilized to find the consensus sequence motifs for various kinases required to catalyze the process of phosphorylation on S/T/Y. These consensus sequences for different kinase groups, families, and individual members are consistent with those described earlier with some novel consensus reported for the first time. A comparison study for the patterns mined by MAPRes with the results of existing prediction methods was performed by searching for these patterns in the vicinity of phosphorylation sites predicted by different available method. This comparison resulted in 87-98% conformity with the results of the predictions by available methods. Additionally, the patterns mined by MAPRes for substrate sites included 61 kinases, the highest number analyzed so far. PMID- 19544400 TI - Osteoprotegerin production by breast cancer cells is suppressed by dexamethasone and confers resistance against TRAIL-induced apoptosis. AB - Osteoprotegerin (OPG) is a decoy receptor for receptor activator of NF-kappaB ligand (RANKL) and TNF-related apoptosis-inducing ligand (TRAIL). While RANKL is essential for osteoclastogenesis and facilitates breast cancer migration into bone, TRAIL promotes breast cancer apoptosis. We analyzed the expression of OPG and TRAIL and its modulation in estrogen receptor-positive MCF-7 cells and receptor-negative MDA-MB-231 cells. In both cells, OPG mRNA levels and protein secretion were dose- and time-dependently enhanced by interleukin (IL)-1beta and suppressed by dexamethasone. In contrast to MCF-7 cells, MDA-MB-231 abundantly expressed TRAIL mRNA, which was enhanced by IL-1beta and inhibited by dexamethasone. TRAIL activated pro-apoptotic caspase-3, -7, and poly-ADP-ribose polymerase and decreased cell numbers of MDA-MB-231, but had no effect on MCF-7 cells. Gene silencing siRNA directed against OPG resulted in a 31% higher apoptotic rate compared to non-target siRNA-treated MDA-MB-231 cells. Furthermore, TRAIL induced significantly less apoptosis in cells cultured in conditioned media (containing OPG) compared to cells exposed to TRAIL in fresh medium lacking OPG (P < 0.01) and these protective effects were reversed by blocking OPG with its specific ligand RANKL (P < 0.05). The association between cancer cell survival and OPG production by MDA-MB-231 cells was further supported by the finding, that modulation of OPG secretion using IL-1beta or dexamethasone prior to TRAIL exposure resulted in decreased and increased rate of apoptosis, respectively (P < 0.05). Thus, OPG secretion by breast cancer cells is modulated by cytokines and dexamethasone, and may represent a critical resistance mechanism that protects against TRAIL-induced apoptosis. PMID- 19544401 TI - Calcium-dependent flagellar motility activation in Chlamydomonas reinhardtii in response to mechanical agitation. AB - Flagellar beating in Chlamydomonas was found to be activated by mechanical stimulation. Immediately after a wild-type cell suspension was vortexed, the average swimming velocity of cells increased from 130 mum/second to 150 mum/second, due to an elevation of flagellar beat frequency from approximately 60 Hz to approximately 70 Hz without detectable change in the flagellar waveforms. This response required outer arm dynein. Treatment with EGTA, Ca(2+)-channel blockers, or mechanosensitive-channel blockers inhibited it. In demembranated and reactivated cell models, a modest increase in Ca(2+) concentration elevated the axonemal beat frequency. These data indicate that the mechanical agitation increases beat frequency because it causes Ca(2+) influx into flagella, which then activates outer arm dynein. PMID- 19544402 TI - Graded actin filament polarity is the organization of oriented actomyosin II filament bundles required for fibroblast polarization. AB - Actomyosin II filament assemblies in cells are required for shaping the cell body and forming the cell rear during morphological polarization and triggering of migration. However, precise steps in myosin II-based mechanisms are unknown in this event; one reason is due to lack of information on the organization of the actin filament substrate for myosin II. Whilst muscle sarcomeric-like contraction drives cell tension in stationary nonmuscle cells, alternative nonsarcomeric modes of myosin II force-generation power forwards movement of the cell body in already migrating cells. Which one contributes to initial cell shape change has not previously been experimentally sought in any polarizing cell. Sarcomeric and nonsarcomeric-based force require completely different types of organization and filament polarity in the actin substrate for myosin II, and these can only currently be distinguished by labour-intensive submicron analysis and electron microscopy. For the first time in any polarizing cell using such analysis we have identified that oriented actomyosin II filament bundles, required for fibroblast polarization, are nonsarcomeric and are organized with graded filament polarity. As this actin organization is similar to the organization in already migrating fibroblasts, we conclude that graded filament polarity is a pivotal myosin II substrate coordinating initial cell shape change and triggering of migration. PMID- 19544403 TI - Connexins, cell motility, and the cytoskeleton. AB - Connexins (Cx) comprise a family of transmembrane proteins, which form intercellular channels between plasma membranes of two adjoining cells, commonly known as gap junctions. Recent reports revealed that Cx proteins interact with diverse cellular components to form a multiprotein complex, which has been termed "Nexus". Potential interaction partners include proteins such as cytoskeletal proteins, scaffolding proteins, protein kinases and phosphatases. These interactions allow correct subcellular localization of Cxs and functional regulation of gap junction-mediated intercellular communication. Evidence is accruing that Cxs might have channel-independent functions, which potentially include regulation of cell migration, cell polarization and growth control. In the current review, we summarize recent knowledge on Cx interactions with cytoskeletal proteins and highlight some aspects of their role in cellular motility. PMID- 19544404 TI - Balancing work and life: a conversation with George Daley. Interviewed by Majlinda Lako and Susan Daher. PMID- 19544405 TI - Balancing work and life: a conversation with Marella de Bruijn. Interviewed by Majlinda Lako and Susan Daher. PMID- 19544406 TI - Medicine on the fringe: stem cell-based interventions in advance of evidence. AB - Stem cell-based interventions (SCBIs) offer great promise; however, there is currently little internationally accepted, scientific evidence supporting the clinical use of SCBIs. The consensus within the scientific community is that a number of hurdles still need to be cleared. Despite this, SCBIs are currently being offered to patients. This article provides a content analysis of materials obtained from SCBI providers. We find content that strains credulity and almost no evidence of SCBIs being delivered in the context of clinical trials. We conclude that until scientific evidence is available, as a general rule, providers should only offer SCBIs in the context of controlled clinical trials. Clients should be aware that the risks and benefits of SCBIs are unknown, that their participation is unlikely to advance scientific knowledge, and they are likely to become ineligible to participate in future clinical trials of SCBIs. We recommend steps to promote patient education and enhance global oversight. PMID- 19544407 TI - Nanog regulates proliferation during early fish development. AB - Nanog is involved in controlling pluripotency and differentiation of stem cells in vitro. However, its function in vivo has been studied only in mouse embryos and various reports suggest that Nanog may not be required for the regulation of differentiation. To better understand endogenous Nanog function, more animal models should be introduced to complement the murine model. Here, we have identified the homolog of the mammalian Nanog gene in teleost fish and describe the endogenous expression of Ol-Nanog mRNA and protein during medaka (Oryzias latipes) embryonic development and in the adult gonads. Using medaka fish as a vertebrate model to study Nanog function, we demonstrate that Ol-Nanog is necessary for S-phase transition and proliferation in the developing embryo. Moreover, inhibition or overexpression of Ol-Nanog does not affect gene expression of various pluripotency and differentiation markers, suggesting that this transcription factor may not play a direct role in embryonic germ layer differentiation. STEM CELLS 2009;27:2081-2091. PMID- 19544408 TI - Abrogation of E-cadherin-mediated cell-cell contact in mouse embryonic stem cells results in reversible LIF-independent self-renewal. AB - We have previously demonstrated that differentiation of embryonic stem (ES) cells is associated with downregulation of cell surface E-cadherin. In this study, we assessed the function of E-cadherin in mouse ES cell pluripotency and differentiation. We show that inhibition of E-cadherin-mediated cell-cell contact in ES cells using gene knockout (Ecad(-/-)), RNA interference (EcadRNAi), or a transhomodimerization-inhibiting peptide (CHAVC) results in cellular proliferation and maintenance of an undifferentiated phenotype in fetal bovine serum-supplemented medium in the absence of leukemia inhibitory factor (LIF). Re expression of E-cadherin in Ecad(-/-), EcadRNAi, and CHAVC-treated ES cells restores cellular dependence to LIF supplementation. Although reversal of the LIF independent phenotype in Ecad(-/-) ES cells is dependent on the beta-catenin binding domain of E-cadherin, we show that beta-catenin null (betacat(-/-)) ES cells also remain undifferentiated in the absence of LIF. This suggests that LIF independent self-renewal of Ecad(-/-) ES cells is unlikely to be via beta-catenin signaling. Exposure of Ecad(-/-), EcadRNAi, and CHAVC-treated ES cells to the activin receptor-like kinase inhibitor SB431542 led to differentiation of the cells, which could be prevented by re-expression of E-cadherin. To confirm the role of transforming growth factor beta family signaling in the self-renewal of Ecad(-/-) ES cells, we show that these cells maintain an undifferentiated phenotype when cultured in serum-free medium supplemented with Activin A and Nodal, with fibroblast growth factor 2 required for cellular proliferation. We conclude that transhomodimerization of E-cadherin protein is required for LIF dependent ES cell self-renewal and that multiple self-renewal signaling networks subsist in ES cells, with activity dependent upon the cellular context. PMID- 19544410 TI - Human bone marrow-derived mesenchymal stromal cells expressing S-TRAIL as a cellular delivery vehicle for human glioma therapy. AB - Glioblastoma is among the most aggressive and treatment resistant of all human cancers. Conventional therapeutic approaches are unsuccessful because of diffuse infiltrative invasion of glioma tumor cells into normal brain parenchyma. Stem cell-based therapies provide a promising approach for the treatment of malignant gliomas because of their migratory ability to invasive tumor cells. Our therapeutic strategy was to use human bone marrow-derived mesenchymal stromal cells (hMSCs) as a cellular vehicle for the targeted delivery and local production of the biologic agent tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) at the glioma tumor site. hMSCs were transduced with a lentivirus expressing secretable TRAIL (S-TRAIL) and mCherry (red fluorescent protein). Our results clearly demonstrate the retention of tumor tropic ability of hMSC S-TRAIL cells by in vitro and in vivo migration assays. In vitro assays confirmed the expression, release, and biological activity of S-TRAIL produced by hMSC S-TRAIL cells. For the in vivo assessment of therapeutic efficacy, hMSCs were injected ipsilateral to an established intracranial glioma tumor in a mouse xenograft model. Genetically engineered hMSC S-TRAIL cells were effective in inhibiting intracranial U87 glioma tumor growth (81.6%) in vivo and resulted in significantly longer animal survival. Immunohistochemical studies demonstrated significant, eight fold greater tumor cell apoptosis in the hMSC S-TRAIL-treated group than in controls. Our study demonstrates the therapeutic efficacy of hMSC S TRAIL cells and confirms that hMSCs can serve as a powerful cell-based delivery vehicle for the site-specific release of therapeutic proteins. PMID- 19544409 TI - High aldehyde dehydrogenase activity: a novel functional marker of murine prostate stem/progenitor cells. AB - We have shown previously that prostatic stem/progenitor cells can be purified from isolated prostate ducts, based on their high expression of the Sca-1 surface antigen. We now report that high levels of aldehyde dehydrogenase (ALDH) activity are present in a subset of prostate epithelial cells that coexpress a number of antigens found on stem/progenitor cells of other origins (CD9, Bcl-2, CD200, CD24, prominin, Oct 3/4, ABCG2, and nestin). Almost all of these cells expressing high levels of ALDH activity also express Sca-1 and a third of them express high levels of this antigen. The cells with high levels of ALDH activity have greater in vitro proliferative potential than cells with low ALDH activity. Importantly, in an in vivo prostate reconstitution assay, the cells expressing high levels of ALDH activity were much more effective in generating prostatic tissue than a population of cells with low enzymatic activity. Thus, a high level of ALDH activity can be considered a functional marker of prostate stem/progenitor cells and allows for simple, efficient isolation of cells with primitive features. The elucidation of the role of ALDH in prostate stem/progenitor cells may lead to the development of rational therapies for treating prostate cancer and benign prostatic hyperplasia. PMID- 19544411 TI - Cell adhesion and spreading affect adipogenesis from embryonic stem cells: the role of calreticulin. AB - Calreticulin is an endoplasmic reticulum-resident multifunctional protein, which has been shown to influence numerous cellular processes, including cell adhesion. In this study, we characterized the adhesive properties of embryonic stem cells (ESCs) lacking calreticulin and showed that adipogenesis from ESCs is directly and reciprocally controlled by the adhesive status of a cell, which in turn is modulated by calreticulin. Calreticulin-deficient ESCs are not only highly adipogenic but also show elevated calmodulin/CaMKII signaling and poor adhesiveness compared with the wild-type ESCs. Calreticulin deficiency leads to a disorganized cytoskeleton and low levels of focal adhesion-related proteins, such as vinculin, paxillin, and phosphorylated focal adhesion kinase, which cause limited focal adhesion formation and limited fibronectin deposition. Moreover, differentiation on nonadhesive substrata, which hinder cell spreading, promoted adipogenesis in the wild-type ESCs that normally have low adipogenic potential, causing a decrease in focal adhesion protein expression and an increase in calmodulin/CaMKII signaling. In contrast, inhibition of CaMKII effectively increased focal adhesion protein levels and inhibited adipogenesis in calreticulin-deficient ESCs, causing them to behave like the low adipogenic, wild type ESCs. Thus, the adipogenic potential of ESCs is proportional to their calmodulin/CaMKII activity but is inversely related to their focal adhesion protein levels and degree of adhesiveness/spreading. PMID- 19544412 TI - Implanted adult human dental pulp stem cells induce endogenous axon guidance. AB - The human central nervous system has limited capacity for regeneration. Stem cell based therapies may overcome this through cellular mechanisms of neural replacement and/or through molecular mechanisms, whereby secreted factors induce change in the host tissue. To investigate these mechanisms, we used a readily accessible human cell population, dental pulp progenitor/stem cells (DPSCs) that can differentiate into functionally active neurons given the appropriate environmental cues. We hypothesized that implanted DPSCs secrete factors that coordinate axon guidance within a receptive host nervous system. An avian embryonic model system was adapted to investigate axon guidance in vivo after transplantation of adult human DPSCs. Chemoattraction of avian trigeminal ganglion axons toward implanted DPSCs was mediated via the chemokine, CXCL12, also known as stromal cell-derived factor-1, and its receptor, CXCR4. These findings provide the first direct evidence that DPSCs may induce neuroplasticity within a receptive host nervous system. PMID- 19544413 TI - Human alternatives to fetal bovine serum for the expansion of mesenchymal stromal cells from bone marrow. AB - Mesenchymal stromal cells (MSCs) are promising candidates for novel cell therapeutic applications. For clinical scale manufacturing, human factors from serum or platelets have been suggested as alternatives to fetal bovine serum (FBS). We have previously shown that pooled human serum (HS) and thrombin activated platelet releasate in plasma (tPRP) support the expansion of adipose tissue-derived MSCs. Contradictory results with bone marrow (BM)-derived MSCs have initiated a comprehensive comparison of HS, tPRP, and pooled human platelet lysate (pHPL) and FBS in terms of their impact on MSC isolation, expansion, differentiation, and immunomodulatory activity. In addition to conventional Ficoll density gradient centrifugation, depletion of lineage marker expressing cells (RosetteSep) and CD271+ sorting were used for BM-MSC enrichment. Cells were cultured in medium containing either 10% FBS, HS, tPRP, or pHPL. Colony-forming units and cumulative population doublings were determined, and MSCs were maximally expanded. Although both HS and tPRP comparable to FBS supported isolation and expansion, pHPL significantly accelerated BM-MSC proliferation to yield clinically relevant numbers within the first two passages. MSC quality and functionality including cell surface marker expression, adipogenic and osteogenic differentiation, and immunosuppressive action were similar in MSCs from all culture conditions. Importantly, spontaneous cell transformation was not observed in any of the culture conditions. Telomerase activity was not detected in any of the cultures at any passage. In contrast to previous data from adipose tissue derived MSCs, pHPL was found to be the most suitable FBS substitute in clinical scale BM-MSC expansion. PMID- 19544414 TI - A targeted neuroglial reporter line generated by homologous recombination in human embryonic stem cells. AB - In this study, we targeted Olig2, a basic helix-loop-helix transcription factor that plays an important role in motoneuron and oligodendrocyte development, in human embryonic stem cell (hESC) line BG01 by homologous recombination. One allele of Olig2 locus was replaced by a green fluorescent protein (GFP) cassette with a targeting efficiency of 5.7%. Targeted clone R-Olig2 (like the other clones) retained pluripotency, typical hESC morphology, and a normal parental karyotype 46,XY. Most importantly, GFP expression recapitulated endogenous Olig2 expression when R-Olig2 was induced by sonic hedgehog and retinoic acid, and GFP positive cells could be purified by fluorescence-activated cell sorting. Consistent with previous reports on rodents, early GFP-expressing cells appeared biased to a neuronal fate, whereas late GFP-expressing cells appeared biased to an oligodendrocytic fate. This was corroborated by myoblast coculture, transplantation into the rat spinal cords, and whole genome expression profiling. The present work reports an hESC reporter line generated by homologous recombination targeting a neural lineage-specific gene, which can be differentiated and sorted to obtain pure neural progenitor populations. PMID- 19544415 TI - Exercise increases neural stem cell number in a growth hormone-dependent manner, augmenting the regenerative response in aged mice. AB - The exercise-induced enhancement of learning and memory, and its ability to slow age-related cognitive decline in humans led us to investigate whether running stimulates periventricular (PVR) neural stem cells (NSCs) in aging mice, thereby augmenting the regenerative capacity of the brain. To establish a benchmark of normal aging on endogenous NSCs, we harvested the PVR from serial vibratome sections through the lateral ventricles of juvenile (6-8 weeks), 6-, 12-, 18-, and 24-month-old mice, culturing the cells in the neural colony-forming cell assay. A significant decline in NSC frequency was apparent by 6 months ( approximately 40%), ultimately resulting in a approximately 90% reduction by 24 months. Concurrent with this decline was a progressive loss in regenerative capacity, as reflected by an incomplete repopulation of neurosphere-forming cells following gamma cell irradiation-induced depletion of the PVR. However, voluntary exercise (i.e., 21 days of running) significantly increased NSC frequency in mice > or = 18 months of age, augmenting the regeneration of irradiation-ablated periventricular cells and restoring NSC numbers to youthful levels. Importantly, and consistent with the demonstrated ability of growth hormone (GH) to increase NSC proliferation, and the elevated secretion of GH during exercise, exercise failed to stimulate NSCs in GH receptor-null mice. These findings now provide a novel basis for understanding the ability of exercise to delay the onset and rate of decline in neurodegenerative conditions not typically associated with the hippocampus and suggest that the GH-dependent activation of endogenous NSCs may be effective in reversing or preventing age-related neurodegeneration in humans. PMID- 19544416 TI - Senescent growth arrest in mesenchymal stem cells is bypassed by Wip1-mediated downregulation of intrinsic stress signaling pathways. AB - Human mesenchymal stem cells (hMSCs) have been widely studied as a source of primary adult stem cells for cell therapy because of their multidifferentiation potential; however, the growth arrest (also known as "premature senescence") often found in hMSCs cultured in vitro has been a major obstacle to the in-depth characterization of these cells. In addition, the inability to maintain constant cell growth hampers the development of additional genetic modifications aimed at achieving desired levels of differentiation to specific tissues; however, the molecular mechanisms that govern this phenomenon remain unclear, with the exception of a few studies demonstrating that induction of p16INK4a is responsible for this senescence-like event. Here, we observed that the premature growth arrest in hMSCs occurs in parallel with the induction of p16INK4a, following abrogation of inhibitory phosphorylation of retinoblastoma protein. These stress responses were concurrent with increased formation of reactive oxygen species (ROSs) from mitochondria and increased p38 mitogen-activated protein kinase (MAPK) activity. The introduction of Wip1 (wild-type p53 inducible phosphatase-1), a well-studied stress modulator, significantly lowered p16INK4a expression and led to p38 MAPK inactivation, although it failed to affect the levels of ROSs. Moreover, the suppression of stress responses by Wip1 apparently extended the life span of hMSCs, compared with control conditions, while maintaining their multilineage differentiation potential. Based on these results, we suggest that senescent growth arrest in hMSCs may result from activation of stress signaling pathways and consequent onset of stress responses, due in part to ROS production during prolonged in vitro culture. PMID- 19544417 TI - Ionizing radiation induces ataxia telangiectasia mutated-dependent checkpoint signaling and G(2) but not G(1) cell cycle arrest in pluripotent human embryonic stem cells. AB - Human embryonic stem (ES) cells are highly sensitive to environmental insults including DNA damaging agents, responding with high levels of apoptosis. To understand the response of human ES cells to DNA damage, we investigated the function of the ataxia telangiectasia mutated (ATM) DNA damage signaling pathway in response to gamma-irradiation. Here, we demonstrate for the first time in human ES cells that ATM kinase is phosphorylated and properly localized to the sites of DNA double-strand breaks within 15 minutes of irradiation. Activation of ATM kinase resulted in phosphorylation of its downstream targets: Chk2, p53, and Nbs1. In contrast to murine ES cells, Chk2 and p53 were localized to the nucleus of irradiated human ES cells. We further show that irradiation resulted in a temporary arrest of the cell cycle at the G(2), but not G(1), phase. Human ES cells resumed cycling approximately 16 hours after irradiation, but had a fourfold higher incidence of aberrant mitotic figures compared to nonirradiated cells. Finally, we demonstrate an essential role of ATM in establishing G(2) arrest since inhibition with the ATM-specific inhibitor KU55933 resulted in abolishment of G(2) arrest, evidenced by an increase in the number of cycling cells 2 hours after irradiation. In summary, these results indicate that human ES cells activate the DNA damage checkpoint, resulting in an ATM-dependent G(2) arrest. However, these cells re-enter the cell cycle with prominent mitotic spindle defects. PMID- 19544419 TI - Purinergic signaling regulates cell proliferation of olfactory epithelium progenitors. AB - In the olfactory epithelium (OE) continuous neurogenesis is maintained throughout life. The OE is in direct contact with the external environment, and its cells are constantly exposed to pathogens and noxious substances. To maintain a functional sense of smell the OE has evolved the ability to permanently replenish olfactory receptor neurons and sustentacular cells lost during natural turnover. A cell population residing in the most basal part of the OE, the so-called basal cells (BCs), keep up this highly regulated genesis of new cells. The population of BCs is thought to include both the stem cells of the OE and various progenitor cells. In recent years a number of regulatory factors that positively and/or negatively regulate the proliferation within the OE have been identified, but a thorough comprehension of the complex interplay of these regulatory factors and the role of the different epithelial cell types is still illusive. Combining labeling techniques, immunohistochemistry, electron microscopy, functional calcium imaging, and a bromo-2'-deoxyuridine incorporation assay, we show for the first time that purinergic receptors are expressed in BCs of the OE of larval Xenopus laevis and that nucleotide-induced Ca(2+) signaling in these cells is involved in the regulation of the cell turnover in the OE. Our data contribute to a better understanding of the regulation of the cell turnover in the OE in particular and also of how the proliferation of neuronal progenitor cells is regulated in general. PMID- 19544418 TI - Smad, PI3K/Akt, and Wnt-dependent signaling pathways are involved in BMP-4 induced ESC self-renewal. AB - It is known that bone morphogenetic protein 4 (BMP-4) has a diverse effect on ESCs. However, its precise mechanism in mouse ESCs is not fully understood. We evaluated the effect of BMP-4 on ESC proliferation and its related signal cascades in this study. BMP-4 significantly increased the level of [(3)H] thymidine incorporation in time- (> or =8 hours) and dose- (> or =10 ng/ml) dependent manners. Additionally, BMP-4 increased cyclin D1 and decreased p27(kip1) expression values in a time-dependent manner. The increases in BMP-4 induced [(3)H]-thymidine incorporation and cyclin D1 expression were inhibited by the BMP-4 receptor antagonist noggin. BMP-4 increased Wnt1 expression. Wnt1 expression was attenuated by Smad4 small interfering RNA (siRNA), and BMP-4 induced cyclin D1 expression was inhibited by Smad4 and Wnt1 siRNAs. BMP-4 also activated beta-catenin, which was blocked by Smad4 and Wnt1 siRNAs. In addition, BMP-4 induced Akt phosphorylation. BMP-4-induced beta-catenin activation and cyclin D1 expression were attenuated by phosphatidyl inositol 3-kinase (PI3K) siRNA and Akt inhibitor. Additionally, downregulation of Smad4, Wnt1, and PI3K expression by siRNA decreased the levels of pluripotency marker mRNAs of ESCs, including Oct4, Sox2, and FoxD3. Our results suggested that BMP-4-induced [(3)H] thymidine incorporation was significantly attenuated by Smad4, Wnt1, and PI3K knockdown. In conclusion, BMP-4 contributed to the maintenance of cell proliferation and the pluripotent state by Smad, PI3K/Akt, and Wnt1/beta-catenin in mouse ESCs. PMID- 19544420 TI - Insulin-secreting cells from human eyelid-derived stem cells alleviate type I diabetes in immunocompetent mice. AB - Various attempts have been made to develop stem cell-based therapy to alleviate type I diabetes using animal models. However, it has been a question whether human insulin produced from explanted cells is solely responsible for the normoglycemia of diabetic animals. In this study, we isolated neural crest-like stem cells from the human eyelid fat and examined their therapeutic potentials for diabetes. The human eyelid adipose-derived stem cells (HEACs) displayed characteristics of neural crest cells. Using a two-step culture condition combined with nicotinamide, activin, and/or GLP-1, we differentiated HEACs into insulin-secreting cells and examined in vivo effects of differentiated cells by transplantation experiments. Following differentiation in vitro, HEACs released insulin and c-peptide in a glucose-dependent manner. Upon their transplantation under kidney capsules of streptozotocin-treated immunocompetent mice, we observed normalization of hyperglycemia in 10 of 20 recipient mice until sacrifice after 2 months. Only the human, but not the mouse, insulin and c-peptide were detected in the blood of recipient mice. Removal of the kidneys transplanted with HEACs resulted in a sharp increase of blood glucose level. Removed kidney tissues showed distinct expression of various human genes including insulin, and colocalization of the human insulin and the human nuclear protein in many cells. However, they showed diminished or null expression of some immune-related genes. In conclusion, human insulin alone produced from eyelid-derived stem cells following differentiation into insulin-secreting cells and transplantation could normalize type I diabetes in mice. PMID- 19544421 TI - Identification of nonepithelial multipotent cells in the embryonic olfactory mucosa. AB - Olfactory mucosal (OM) tissue, a potential source of stem cells, is currently being assessed in the clinic as a candidate tissue for transplant-mediated repair of spinal cord injury. We examined the ability of embryonic rat OM tissue to generate stem cells using culture conditions known to promote neural stem cell proliferation. Primary spheres formed that proliferated and exhibited two main morphologies: (a) CNS neurosphere-like (OM-I) and (b) small, tight spheroid-like (OM-II). The OM-I spheres expressed the neural stem cell marker nestin but also markers of peripheral glia, neurons, and connective tissue. Further studies demonstrated the presence of multipotential mesenchymal-like stem cells within OM I spheres that differentiated into bone, adipose, and smooth muscle cells. In contrast, the OM-II spheres contained mainly cytokeratin-expressing cells. Immunolabeling of rat olfactory tissue with Stro-1, CD90, and CD105 showed the presence of multipotent mesenchymal cells in the lamina propria, whereas cytokeratin was expressed by the epithelial cells of the olfactory epithelium. In addition, a comparable pattern of immunoreactivity was detected in human tissue using Stro-1 and cytokeratin, suggesting the presence of similar cells in this tissue. The identification of a nonepithelial multipotent cell in the OM may explain the varied reports on olfactory stem cell differentiation capacity in vitro and in vivo and illustrates the cellular complexity of this tissue as a potential source of stem cells for transplantation and translation to the clinic. PMID- 19544422 TI - CARM1 is required in embryonic stem cells to maintain pluripotency and resist differentiation. AB - Histone H3 methylation at R17 and R26 recently emerged as a novel epigenetic mechanism regulating pluripotency in mouse embryos. Blastomeres of four-cell embryos with high H3 methylation at these sites show unrestricted potential, whereas those with lower levels cannot support development when aggregated in chimeras of like cells. Increasing histone H3 methylation, through expression of coactivator-associated-protein-arginine-methyltransferase 1 (CARM1) in embryos, elevates expression of key pluripotency genes and directs cells to the pluripotent inner cell mass. We demonstrate CARM1 is also required for the self renewal and pluripotency of embryonic stem (ES) cells. In ES cells, CARM1 depletion downregulates pluripotency genes leading to their differentiation. CARM1 associates with Oct4/Pou5f1 and Sox2 promoters that display detectable levels of R17/26 histone H3 methylation. In CARM1 overexpressing ES cells, histone H3 arginine methylation is also at the Nanog promoter to which CARM1 now associates. Such cells express Nanog at elevated levels and delay their response to differentiation signals. Thus, like in four-cell embryo blastomeres, histone H3 arginine methylation by CARM1 in ES cells allows epigenetic modulation of pluripotency. PMID- 19544423 TI - c-Kit function is necessary for in vitro myogenic differentiation of bone marrow hematopoietic cells. AB - In recent years, the differentiation of bone marrow cells (BMCs) into myocytes has been extensively investigated, but the findings remain inconclusive. The purpose of this study was to determine the conditions necessary to induce myogenic differentiation in short-term cultures of adult BMCs, and to identify the BMC subpopulation responsible for this phenomenon. We report that high density cultures of murine hematopoietic BMCs gave rise to spontaneous beating cell clusters in the presence of vascular endothelial and fibroblast growth factors. These clusters originated from c-kit(pos) cells. The formation of the clusters could be completely blocked by adding a c-kit/tyrosine kinase inhibitor, Gleevec (imatinib mesylate; Novartis International, Basel, Switzerland, http://www.novartis.com), to the culture. Cluster formation was also blunted in BMCs from c-kit-deficient (Kit(W)/Kit(W-v)) mice. Clustered cells expressed cardiomyocyte-specific transcription factor genes Gata-4 and Nkx2.5, sarcomeric proteins beta-MHC and MLC-2v, and ANF and connexin-43. Immunostaining revealed alpha-sarcomeric actinin expression in more than 90% of clustered cells. Under electron microscopy, the clustered cells exhibited a sarcomeric myofiber arrangement and z-bands. This study defines the microenvironment required to achieve a reproducible in vitro model of beating, myogenic cell clusters. This model could be used to examine the mechanisms responsible for the postnatal myogenic differentiation of BMCs. Our results identify c-kit(pos) bone marrow hematopoietic cells as the source of the myogenic clusters. PMID- 19544424 TI - Coculture of human embryonic stem cells and human articular chondrocytes results in significantly altered phenotype and improved chondrogenic differentiation. AB - Human embryonic stem (hES) cells have been suggested as a cell source for the repair of cartilage lesions. Here we studied how coculture with human articular chondrocytes affects the expansion potential, morphology, expression of surface markers, and differentiation abilities of hES cells, with special regard to chondrogenic differentiation. Undifferentiated hES cells were cocultured with irradiated neonatal or adult articular chondrocytes in high-density pellet mass cultures for 14 days. Cocultured hES cells were then expanded on plastic and their differentiation potential toward the adipogenic, osteogenic, and chondrogenic lineages was compared with that of undifferentiated hES cells. The expression of different surface markers was investigated using flow cytometry and teratoma formation was studied using injection of the cells under the kidney capsule. Our results demonstrate that although hES cells have to be grown on Matrigel, the cocultured hES cells could be massively expanded on plastic with a morphology and expression of surface markers similar to mesenchymal stem cells. Coculture further resulted in a more homogenous pellet and significantly increased cartilage matrix production, both in high-density pellet mass cultures and hyaluronan-based scaffolds. Moreover, cocultured cells formed colonies in agarose suspension culture, also demonstrating differentiation toward chondroprogenitor cells, whereas no colonies were detected in the hES cell cultures. Coculture further resulted in a significantly decreased osteogenic potential. No teratoma formation was detected. Our results confirm the potential of the culture microenvironment to influence hES cell morphology, expansion potential, and differentiation abilities over several population doublings. PMID- 19544425 TI - Locally delivered growth factor enhances the angiogenic efficacy of adipose derived stromal cells transplanted to ischemic limbs. AB - Ischemia is a potentially fatal medical event that is associated with as many as 30% of all deaths. Stem cell therapy offers significant therapeutic promise, but poor survival following transplantation to ischemic tissue limits its efficacy. Here we demonstrate that nanosphere-mediated growth factor delivery can enhance the survival of transplanted human adipose-derived stromal cells (hADSCs) and secretion of human angiogenic growth factors per cell, and substantially improve therapeutic efficacy of hADSCs. In vitro, in hypoxic (1% oxygen) and serum deprived conditions that simulate in vivo ischemia, fibroblast growth factor-2 (FGF2) significantly reduced hADSC apoptosis and enhanced angiogenic growth factor secretion. In vivo, hADSCs delivered intramuscularly into ischemic hind limbs in combination with FGF2 resulted in significant improvements in limb survival and blood perfusion, as well as survival of the transplanted hADSCs and secretion of human angiogenic growth factors (i.e., vascular endothelial growth factor, hepatocyte growth factor, and FGF2). Interestingly, the majority of transplanted hADSCs were localized adjacent to the microvessels rather than being incorporated into them, suggesting that their major contribution to angiogenesis might be to increase paracrine secretion of angiogenic growth factors. This study demonstrates the potential of hADSCs in combination with growth factors for use in the treatment of ischemia. PMID- 19544426 TI - Generation of pancreatic hormone-expressing islet-like cell aggregates from murine adipose tissue-derived stem cells. AB - The success of cell replacement therapy for diabetes depends on the availability and generation of an adequate number of islets, preferably from an autologous origin. Stem cells are now being probed for the generation of physiologically competent, insulin-producing cells. In this investigation, we explored the potential of adipose tissue-derived stem cells (ASCs) to differentiate into pancreatic hormone-expressing islet-like cell aggregates (ICAs). We initiated ASC culture from epididymal fat pads of Swiss albino mice to obtain mesenchymal cells, murine epididymal (mE)-ASCs. Subsequent single-cell cloning resulted in a homogeneous cell population with a CD29(+)CD44(+)Sca-1(+) surface antigen expression profile. We formulated a 10-day differentiation protocol to generate insulin-expressing ICAs from mE-ASCs by progressively changing the differentiation cocktail on day 1, day 3, and day 5. Our stage-specific approach successfully differentiated mesodermic mE-ASCs into definitive endoderm (cells expressing Sox17, Foxa2, GATA-4, and cytokeratin [CK]-19), then into pancreatic endoderm (cells expressing pancreatic and duodenal homeobox [PDX]-1, Ngn3, NeuroD, Pax4, and glucose transporter 2), and finally into cells expressing pancreatic hormones (insulin, glucagon, somatostatin). Fluorescence-activated cell sorting analysis showed that day 5 ICAs contained 64.84% +/- 7.03% PDX-1(+) cells, and in day 10 mature ICAs, 48.17% +/- 3% of cells expressed C-peptide. Day 10 ICAs released C-peptide in a glucose-dependent manner, exhibiting in vitro functionality. Electron microscopy of day 10 ICAs revealed the presence of numerous secretory granules within the cell cytoplasm. Calcium alginate encapsulated day 10 ICAs (1,000-1,200), when transplanted i.p. into streptozotocin-induced diabetic mice, restored normoglycemia within 2 weeks. The data presented here demonstrate the feasibility of using ASCs as a source of autologous stem cells to differentiate into the pancreatic lineage. PMID- 19544427 TI - Species variation in the mechanisms of mesenchymal stem cell-mediated immunosuppression. AB - Bone marrow-derived mesenchymal stem cells (MSCs) hold great promise for treating immune disorders because of their immunoregulatory capacity, but the mechanism remains controversial. As we show here, the mechanism of MSC-mediated immunosuppression varies among different species. Immunosuppression by human- or monkey-derived MSCs is mediated by indoleamine 2,3-dioxygenase (IDO), whereas mouse MSCs utilize nitric oxide, under the same culture conditions. When the expression of IDO and inducible nitric oxide synthase (iNOS) were examined in human and mouse MSCs after stimulation with their respective inflammatory cytokines, we found that human MSCs expressed extremely high levels of IDO, and very low levels of iNOS, whereas mouse MSCs expressed abundant iNOS and very little IDO. Immunosuppression by human MSCs was not intrinsic, but was induced by inflammatory cytokines and was chemokine-dependent, as it is in mouse. These findings provide critical information about the immunosuppression of MSCs and for better application of MSCs in treating immune disorders. PMID- 19544428 TI - Stem cell transplant into preimplantation embryo yields myocardial infarction resistant adult phenotype. AB - Stem cells are an emerging strategy for treatment of myocardial infarction, limited however to postinjury intervention. Preventive stem cell-based therapy to augment stress tolerance has yet to be considered for lifelong protection. Here, pluripotent stem cells were microsurgically introduced at the blastocyst stage of murine embryo development to ensure stochastic integration and sustained organ contribution. Engineered chimera displayed excess in body weight due to increased fat deposits, but were otherwise devoid of obesity-related morbidity. Remarkably, and in sharp contrast to susceptible nonchimeric offspring, chimera was resistant to myocardial infarction induced by permanent coronary occlusion. Infarcted nonchimeric adult hearts demonstrated progressive deterioration in ejection fraction, while age-matched 12-14-months-old chimera recovered from equivalent ischemic insult to regain within one-month preocclusion contractile performance. Electrical remodeling and ventricular enlargement with fibrosis, prominent in failing nonchimera, were averted in the chimeric cohort characterized by an increased stem cell load in adipose tissue and upregulated markers of biogenesis Ki67, c-Kit, and stem cell antigen-1 in the myocardium. Favorable outcome in infarcted chimera translated into an overall benefit in workload capacity and survival. Thus, prenatal stem cell transplant yields a cardioprotective phenotype in adulthood, expanding cell-based indications beyond traditional postinjury applications to include pre-emptive therapy. PMID- 19544430 TI - Loss of ATM impairs proliferation of neural stem cells through oxidative stress mediated p38 MAPK signaling. AB - Ataxia-telangiectasia (A-T) is a genetic disorder caused by a mutation of the Atm gene, which controls DNA repair, cell cycling, and redox homeostasis. Even though oxidative stress has been implicated in the neurological anomalies in A-T, the effects of ATM loss on neural stem cell (NSC) survival has remained elusive. In this study, we investigated the effects of oxidative stress on NSC proliferation in an animal model for A-T neurodegeneration. We found that cultured subventricular zone neurosphere cells from Atm(-/-) mice show impaired proliferation, as well as intrinsic elevation of reactive oxygen species (ROS) levels, compared with those from Atm(+/+) mice. We also show that increasing the levels of ROS by H(2)O(2) treatment significantly reduces Atm(+/+) neurosphere formation and proliferation. In Atm(-/-) neurosphere cells, the Akt and Erk1/2 pathways are disrupted, together with enhanced activity of the p38 mitogen activated protein kinase (MAPK). Treatment of these cells with the antioxidant N acetyl-L-cysteine (NAC) or with a p38 MAPK inhibitor restores normal proliferation and reduced expression of p21(cip1) and p27(kip1) in the Atm(-/-) NSCs. These observations indicate that ATM plays a crucial role in NSC proliferation, by activating Akt and Erk1/2 pathways and by suppressing ROS-p38 MAPK signaling. Together, our results suggest that p38 MAPK signaling acts as a negative regulator of NSC proliferation in response to oxidative stress. These findings suggest a potential mechanism for neuronal cell loss as a result of oxidative stress in NSCs in progressive neurodegenerative diseases such as A-T. PMID- 19544429 TI - Epidermal growth factor induces the progeny of subventricular zone type B cells to migrate and differentiate into oligodendrocytes. AB - New neurons and oligodendrocytes are continuously produced in the subventricular zone (SVZ) of adult mammalian brains. Under normal conditions, the SVZ primary precursors (type B1 cells) generate type C cells, most of which differentiate into neurons, with a small subpopulation giving rise to oligodendrocytes. Epidermal growth factor (EGF) signaling induces dramatic proliferation and migration of SVZ progenitors, a process that could have therapeutic applications. However, the fate of cells derived from adult neural stem cells after EGF stimulation remains unknown. Here, we specifically labeled SVZ B1 cells and followed their progeny after a 7-day intraventricular infusion of EGF. Cells derived from SVZ B1 cells invaded the parenchyma around the SVZ into the striatum, septum, corpus callosum, and fimbria-fornix. Most of these B1-derived cells gave rise to cells in the oligodendrocyte lineage, including local NG2+ progenitors, and pre-myelinating and myelinating oligodendrocytes. SVZ B1 cells also gave rise to a population of highly-branched S100beta+/glial fibrillary acidic protein (GFAP)+ cells in the striatum and septum, but no neuronal differentiation was observed. Interestingly, when demyelination was induced in the corpus callosum by a local injection of lysolecithin, an increased number of cells derived from SVZ B1 cells and stimulated to migrate and proliferate by EGF infusion differentiated into oligodendrocytes at the lesion site. This work indicates that EGF infusion can greatly expand the number of progenitors derived from the SVZ primary progenitors which migrate and differentiate into oligodendroglial cells. This expanded population could be used for the repair of white matter lesions. PMID- 19544431 TI - A complex role for FGF-2 in self-renewal, survival, and adhesion of human embryonic stem cells. AB - The transcription program that is responsible for the pluripotency of human ESCs (hESCs) is believed to be comaintained by exogenous fibroblast growth factor-2 (FGF-2), which activates FGF receptors (FGFRs) and stimulates the mitogen activated protein kinase (MAPK) pathway. However, the same pathway is stimulated by insulin receptors, insulin-like growth factor 1 receptors, and epidermal growth factor receptors. This mechanism is further complicated by intracrine FGF signals. Thus, the molecular mechanisms by which FGF-2 promotes the undifferentiated growth of hESCs are unclear. Here we show that, in undifferentiated hESCs, exogenous FGF-2 stimulated the expression of stem cell genes while suppressing cell death and apoptosis genes. Inhibition of autocrine FGF signaling caused upregulation of differentiation-related genes and downregulation of stem cell genes. Thus, exogenous FGF-2 reinforced the pluripotency maintenance program of intracrine FGF-2 signaling. Consistent with this hypothesis, expression of endogenous FGF-2 decreased during hESC differentiation and FGF-2 knockdown-induced hESC differentiation. In addition, FGF-2 signaling via FGFR2 activated MAPK kinase/extracellular signal-regulated kinase and AKT kinases, protected hESC from stress-induced cell death, and increased hESC adhesion and cloning efficiency. This stimulation of self-renewal, cell survival, and adhesion by exogenous and endogenous FGF-2 may synergize to maintain the undifferentiated growth of hESCs. PMID- 19544432 TI - EpCAM is involved in maintenance of the murine embryonic stem cell phenotype. AB - Epithelial cell adhesion molecule EpCAM is a transmembrane glycoprotein that is expressed on subsets of normal epithelia, numerous stem- and progenitor-type cells, and most carcinomas and highly overexpressed on cancer-initiating cells. The role of EpCAM in early development, particularly in stem-like cells, has remained unclear. Here, we show that the maintenance of self-renewal in murine embryonic stem (ES) cells depends on the high-level expression of EpCAM. Cultivation of ES cells under differentiation conditions in the absence of leukemia inhibitory factor (LIF) caused down-regulation of EpCAM along with decreased expression of cellular myelocytomatosis oncogene (c-Myc), Sex determining region Y-Box 2, Octamer 3/4 (Oct3/4), and Stat3. As a consequence ES cells were morphologically differentiated and ceased to proliferate. RNA interference-mediated inhibition of EpCAM expression under self-renewal conditions resulted in quantitatively decreased proliferation, decreased Oct3/4, SSEA-1, and c-Myc expression, and diminished alkaline phosphatase activity. Conversely, exogenous expression of EpCAM partially compensated for the requirement of ES cells for LIF to retain a stem cell phenotype. Thus, murine EpCAM is a transmembrane protein, which is essential but by itself is not sufficient for maintenance of the ES cell phenotype. PMID- 19544433 TI - Proliferation of human glioblastoma stem cells occurs independently of exogenous mitogens. AB - Primary glial tumors of the central nervous system, most commonly glioblastoma multiforme (GBM), are aggressive lesions with a dismal prognosis. Despite identification and isolation of human brain tumor stem cells (BTSCs), characteristics that distinguish BTSCs from neural stem cells remain to be elucidated. We cultured cells isolated from gliomas, using the neurosphere culture system, to understand their growth requirements. Both CD133(+) and CD133( ) adult GBM BTSCs proliferated in the absence of exogenous mitogenic stimulation and gave rise to multipotent GBM spheres that were capable of self-renewal. Epidermal growth factor (EGF) and fibroblast growth factor-2 enhanced GBM BTSC survival, proliferation, and subsequent sphere size. Blockade of EGF receptor (EGFR) signaling reduced exogenous mitogen-independent GBM sphere growth. Implantation of as few as 10 exogenous mitogen-independent GBM BTSCs led to the formation of highly invasive intracranial tumors, which closely resembled human GBMs, in immunocompromised mice. These results demonstrate that exogenous mitogen independence, mediated in part through EGFR signaling, is one characteristic that distinguishes CD133(+) and CD133(-) GBM BTSCs from neural stem cells. This novel experimental system will permit the elucidation of additional constitutively activated mechanisms that promote GBM BTSC survival, self-renewal, and proliferation. PMID- 19544435 TI - mAb 84, a cytotoxic antibody that kills undifferentiated human embryonic stem cells via oncosis. AB - The monoclonal antibody mAb 84, which binds to podocalyxin-like protein-1 (PODXL) on human embryonic stem cells (hESCs), was previously reported to bind and kill undifferentiated cells in in vitro and in vivo assays. In this study, we investigate the mechanism responsible for mAb 84-induced hESCs cytotoxicity. Apoptosis was likely not the cause of mAb 84-mediated cell death because no elevation of caspase activities or increased DNA fragmentation was observed in hESCs following incubation with mAb 84. Instead, it was preceded by cell aggregation and damage to cell membranes, resulting in the uptake of propidium iodide, and the leakage of intracellular sodium ions. Furthermore, examination of the cell surface by scanning electron microscopy revealed the presence of pores on the cell surface of mAb 84-treated cells, which was absent from the isotype control. This mechanism of cell death resembles that described for oncosis, a form of cell death resulting from membrane damage. Additional data suggest that the binding of mAb 84 to hESCs initiates a sequence of events prior to membrane damage, consistent with oncosis. Degradation of actin-associated proteins, namely, alpha-actinin, paxillin, and talin, was observed. The perturbation of these actin-associated proteins consequently permits the aggregation of PODXL, thus leading to the formation of pores. To our knowledge, this is the first report of oncotic cell death with hESCs as a model. PMID- 19544434 TI - Regulation of neural specification from human embryonic stem cells by BMP and FGF. AB - Inhibition of bone morphogenetic protein (BMP) signaling is required for vertebrate neural induction, and fibroblast growth factors (FGFs) may affect neural induction through phosphorylation at the linker region of Smad1, thus regulating BMP signaling. Here we show that human embryonic stem cells efficiently convert to neuroepithelial cells in the absence of BMP antagonists, or even when exposed to high concentrations of exogenous BMP4. Molecular and functional analyses revealed multiple levels of endogenous BMP signaling inhibition that may account for the efficient neural differentiation. Blocking FGF signaling inhibited neural induction, but did not alter the phosphorylation of the linker region of Smad1, suggesting that FGF enhances human neural specification independently of BMP signaling. PMID- 19544436 TI - Efficient adipocyte and osteoblast differentiation from mouse induced pluripotent stem cells by adenoviral transduction. AB - Induced pluripotent stem (iPS) cells, which are generated from somatic cells by transducing four genes, are expected to have broad application to regenerative medicine. Although establishment of an efficient gene transfer system for iPS cells is considered to be essential for differentiating them into functional cells, the detailed transduction characteristics of iPS cells have not been examined. Previously, by using an adenovirus (Ad) vector containing the elongation factor-1alpha (EF-1alpha) and the cytomegalovirus enhancer/beta-actin (CA) promoters, we developed an efficient transduction system for mouse embryonic stem (ES) cells and their aggregate form, embryoid bodies (EBs). In this study, we applied our transduction system to mouse iPS cells and investigated whether efficient differentiation could be achieved by Ad vector-mediated transduction of a functional gene. As in the case of ES cells, the Ad vector containing EF-1alpha and the CA promoter could efficiently transduce transgenes into mouse iPS cells. At 3,000 vector particles/cell, 80%-90% of iPS cells expressed transgenes by treatment with an Ad vector containing the CA promoter, without a decrease in pluripotency or viability. We also found that the CA promoter had potent transduction ability in iPS cell-derived EBs. Moreover, exogenous expression of a PPARgamma gene or a Runx2 gene into mouse iPS cells by an optimized Ad vector enhanced adipocyte or osteoblast differentiation, respectively. These results suggest that Ad vector-mediated transient transduction is sufficient to increase cellular differentiation and that our transduction methods would be useful for therapeutic applications based on iPS cells. PMID- 19544437 TI - Nicotinamide rescues human embryonic stem cell-derived neuroectoderm from parthanatic cell death. AB - Abundant cell death is observed when human embryonic stem cells (hESCs) undergo neuralization, a critical first step for future cell-based therapies addressing neurodegeneration. Using hESC neuralization as an in vitro model of human development, we demonstrated that the developing neuroepithelium acquires increased susceptibility to spontaneous cell death. We found that poly(ADP ribose) polymerase-1 (PARP1)/apoptosis-inducing factor (AIF)-mediated cell death (parthanatos) is a dominant mechanism responsible for cell loss during hESC neuralization. The demise of neural progenitor cells, at least in part, is due to decreased endogenous antioxidant defenses and enhanced reactive oxygen species leakage from mitochondria fuelled by nonphysiological culture conditions. Under such conditions, PARP1 overactivation triggered cell death through the mitochondrial-nuclear translocation of AIF. Blocking PARP1 activity with small hairpin RNA interference or nicotinamide dramatically enhanced hESC neuralization, providing optimal survival of the developing neuroepithelium. Because nicotinamide is a physiological metabolite, our results raise the possibility that neural stem/progenitor cell survival in vivo requires a metabolic niche. We argue that small natural metabolites provide a powerful physiological tool to optimize hESC differentiation compatible with the requirements of regenerative medicine. PMID- 19544438 TI - Microgel iron oxide nanoparticles for tracking human fetal mesenchymal stem cells through magnetic resonance imaging. AB - Stem cell transplantation for regenerative medicine has made significant progress in various injury models, with the development of modalities to track stem cell fate and migration post-transplantation being currently pursued rigorously. Magnetic resonance imaging (MRI) allows serial high-resolution in vivo detection of transplanted stem cells labeled with iron oxide particles, but has been hampered by low labeling efficiencies. Here, we describe the use of microgel iron oxide (MGIO) particles of diameters spanning 100-750 nm for labeling human fetal mesenchymal stem cells (hfMSCs) for MRI tracking. We found that MGIO particle uptake by hfMSCs was size dependent, with 600-nm MGIO (M600) particles demonstrating three- to sixfold higher iron loading than the clinical particle ferucarbotran (33-263 versus 9.6-42.0 pg iron/hfMSC; p < .001). Cell labeling with either M600 particles or ferucarbotran did not affect either cellular proliferation or tri-lineage differentiation into osteoblasts, adipocytes, and chondrocytes, despite differences in gene expression on a genome-wide microarray analysis. Cell tracking in a rat photothrombotic stroke model using a clinical 1.5-T MRI scanner demonstrated the migration of labeled hfMSCs from the contralateral cortex to the stroke injury, with M600 particles achieving a five- to sevenfold higher sensitivity for MRI detection than ferucarbotran (p < .05). However, model-related cellular necrosis and acute inflammation limited the survival of hfMSCs beyond 5-12 days. The use of M600 particles allowed high detection sensitivity with low cellular toxicity to be achieved through a simple incubation protocol, and may thus be useful for cellular tracking using standard clinical MRI scanners. PMID- 19544439 TI - Noninvasive bioluminescent imaging demonstrates long-term multilineage engraftment of ex vivo-expanded CD34-selected umbilical cord blood cells. AB - The use of umbilical cord blood (UCB) grafts for hematopoietic stem cell transplantation (HSCT) is a promising technique that permits a degree of human leukocyte antigen mismatch between the graft and the host without the concomitant higher rate of graft-versus-host disease that would be observed between an adult marrow graft and a mismatched host. A disadvantage to the use of UCB for HSCT is that immune reconstitution may be significantly delayed because of the low stem cell dose available in the graft. Ex vivo expansion of UCB CD34 cells would provide a greater number of stem cells; however, there are persistent concerns that ex vivo-expanded CD34 cells may lose pluripotency and the ability to contribute meaningfully to long-term engraftment. To address this issue, we transduced CD34-selected UCB cells with a lentiviral construct expressing luciferase, and determined homing and engraftment patterns in vivo by noninvasive bioluminescent imaging in sublethally irradiated NOD/SCID/IL-2Rgamma(-/-) (NSG) mice. Graft contribution to multilineage commitment was also confirmed by analysis of primary and secondary transplants by flow cytometry and immunohistochemistry. Our results demonstrate that, other than a mild delay at the onset of engraftment, there were no significant differences in lineage repopulation or in long-term or secondary engraftment between culture-expanded and unexpanded UCB CD34-selected cells. The results suggest that multipotent stem cells can be expanded ex vivo and can contribute meaningfully to long-term hematopoietic engraftment. PMID- 19544440 TI - Novel STAT3 target genes exert distinct roles in the inhibition of mesoderm and endoderm differentiation in cooperation with Nanog. AB - Leukemia inhibitory factor (LIF) activates the transcription factor signal transducer and activator of transcription 3 (STAT3), which results in the maintenance of mouse embryonic stem cells in the pluripotent state by inhibiting both mesodermal and endodermal differentiation. How the LIF/STAT3 pathway inhibits commitment to both mesoderm and endoderm lineages is presently unknown. Using a hormone-dependent STAT3 and with microarray analysis, we identified 58 targets of STAT3 including 20 unknown genes. Functional analysis showed that 22 among the 23 STAT3 target genes analyzed contribute to the maintenance of the undifferentiated state, as evidenced by an increase in the frequency of differentiated colonies in a self-renewal assay and a concomitant elevation of early differentiation markers upon knockdown. Fourteen of them, including Dact1, Klf4, Klf5, Rgs16, Smad7, Ccrn4l, Cnnm1, Ocln, Ier3, Pim1, Cyr61, and Sgk, were also regulated by Nanog. Analysis of lineage-specific markers showed that the STAT3 target genes fell into three distinct categories, depending on their capacity to inhibit either mesoderm or endoderm differentiation or both. The identification of genes that harness self-renewal and are downstream targets of both STAT3 and Nanog shed light on the mechanisms underlying functional redundancy between STAT3 and Nanog in mouse embryonic stem cells. PMID- 19544441 TI - Direct transdifferentiation of stem/progenitor spermatogonia into reproductive and nonreproductive tissues of all germ layers. AB - Pluripotent stem cells have great clinical potential for tissue regeneration/repair in humans. The use of embryonic stem (ES) cells is ethically controversial, leading to searches for other sources of pluripotent stem cells. Testicular spermatogonial stem cells (SSCs) produce the spermatogenic lineage. Under in vitro conditions, SSCs have the ability to give rise to pluripotent ES like cells. We hypothesized that stem/progenitor spermatogonia could directly transdifferentiate into different tissue types if they were recombined with inductive mesenchymes from fetal/neonatal organs using a tissue separation/recombination methodology and grown in vivo. Green fluorescent protein transgenic mice were used to track cell lineages. Our results indicate that stem/progenitor spermatogonia recombined with the appropriate mesenchyme can directly transdifferentiate in vivo into tissues of all germ layers, including prostatic, uterine, and skin epithelium. In addition, transdifferentiated tissue expressed molecular, histological, and functional markers of the appropriate epithelium. The ability of stem/progenitor spermatogonia to directly generate various epithelia emphasizes their clinical potential, and if adult human SSCs have similar properties, this may have applications in human regenerative medicine. PMID- 19544442 TI - Notch increases T/NK potential of human hematopoietic progenitors and inhibits B cell differentiation at a pro-B stage. AB - Notch and its ligands regulate multiple cell fate decisions. However, several questions on the timing, durability, and reversibility of Notch signaling effects on human hematopoietic precursors are still unresolved. Here, we used recombinant Delta ligands to deliver temporally and dose-controlled signals to human immature cord blood CD34(+)CD38(low) cells at clonal cell levels. Notch activation increased the frequency of multipotent progenitors, skewed the T and natural killer (NK) cell potential of CD34(+)CD38(low) clones in a dose- and ligand dependent manner, and inhibited the differentiation of B cell clones. Low doses of ligands were sufficient for significantly increasing the frequency of NK cell precursors, whereas higher doses were required for increasing the frequency of T cell clones. Interestingly, we demonstrate that temporary Notch activation prevents the subsequent differentiation of CD34(+)CD38(low) cells beyond a pro-B CD79a(+)CD19(-) stage characterized as a common lymphoid progenitor (CLP). Moreover, the lymphoid potential of this pro-B/CLP was skewed toward NK cell potential while the B cell precursor frequency was dramatically reduced. These results indicate critical timing and quantitative aspects of Notch/Delta interactions, imprinting the potential of CD34(+)CD38(low) hematopoietic progenitors. These results may have implications both in physiology and for cell manipulation because they demonstrate a tight regulation of the fate of human progenitors by Notch signaling. PMID- 19544443 TI - A boost of BMP4 accelerates the commitment of human embryonic stem cells to the endothelial lineage. AB - Embryoid bodies (EBs) generated during differentiation of human embryonic stem cells (hESCs) contain vascular-like structures, suggesting that commitment of mesoderm progenitors into endothelial cells occurs spontaneously. We showed that bone morphogenetic protein 4 (BMP4), an inducer of mesoderm, accelerates the peak expression of CD133/kinase insert domain-containing receptor (KDR) and CD144/KDR. Because the CD133(+)KDR(+) population could represent endothelial progenitors, we sorted them at day 7 and cultured them in endothelial medium. These cells were, however, unable to differentiate into endothelial cells. Under standard conditions, the CD144(+)KDR(+) population represents up to 10% of the total cells at day 12. In culture, these cells, if sorted, give rise to a homogeneous population with a morphology typical of endothelial cells and express endothelial markers. These endothelial cells derived from the day 12 sorted population were functional, as assessed by different in vitro assays. When EBs were stimulated by BMP4, the CD144(+)KDR(+) peak was shifted to day 7. Most of these cells, however, were CD31(-), becoming CD31(+) in culture. They then expressed von Willebrand factor and were functional. This suggests that, initially, the BMP4-boosted day 7, CD144(+)KDR(+)CD31(-) population represents immature endothelial cells that differentiate into mature endothelial cells in culture. The expression of OCT3/4, a marker of immaturity for hESCs decreases during EB differentiation, decreasing faster following BMP4 induction. We also show that BMP4 inhibits the global expression of GATA2 and RUNX1, two transcription factors involved in hemangioblast formation, at day 7 and day 12. PMID- 19544444 TI - miR-200 regulates PDGF-D-mediated epithelial-mesenchymal transition, adhesion, and invasion of prostate cancer cells. AB - MicroRNAs have been implicated in tumor progression. Recent studies have shown that the miR-200 family regulates epithelial-mesenchymal transition (EMT) by targeting zinc-finger E-box binding homeobox 1 (ZEB1) and ZEB2. Emerging evidence from our laboratory and others suggests that the processes of EMT can be triggered by various growth factors, such as transforming growth factor beta and platelet-derived growth factor-D (PDGF-D). Moreover, we recently reported that overexpression of PDGF-D in prostate cancer cells (PC3 PDGF-D cells) leads to the acquisition of the EMT phenotype, and this model offers an opportunity for investigating the molecular interplay between PDGF-D signaling and EMT. Here, we report, for the first time, significant downregulation of the miR-200 family in PC3 PDGF-D cells as well as in PC3 cells exposed to purified active PDGF-D protein, resulting in the upregulation of ZEB1, ZEB2, and Snail2 expression. Interestingly, re-expression of miR-200b in PC3 PDGF-D cells led to reversal of the EMT phenotype, which was associated with the downregulation of ZEB1, ZEB2, and Snail2 expression, and these results were consistent with greater expression levels of epithelial markers. Moreover, transfection of PC3 PDGF-D cells with miR 200b inhibited cell migration and invasion, with concomitant repression of cell adhesion to the culture surface and cell detachment. From these results, we conclude that PDGF-D-induced acquisition of the EMT phenotype in PC3 cells is, in part, a result of repression of miR-200 and that any novel strategy by which miR 200 could be upregulated would become a promising approach for the treatment of invasive prostate cancer. PMID- 19544445 TI - Regenerative effects of transplanted mesenchymal stem cells in fracture healing. AB - Mesenchymal stem cells (MSC) have a therapeutic potential in patients with fractures to reduce the time of healing and treat nonunions. The use of MSC to treat fractures is attractive for several reasons. First, MSCs would be implementing conventional reparative process that seems to be defective or protracted. Secondly, the effects of MSCs treatment would be needed only for relatively brief duration of reparation. However, an integrated approach to define the multiple regenerative contributions of MSC to the fracture repair process is necessary before clinical trials are initiated. In this study, using a stabilized tibia fracture mouse model, we determined the dynamic migration of transplanted MSC to the fracture site, their contributions to the repair process initiation, and their role in modulating the injury-related inflammatory responses. Using MSC expressing luciferase, we determined by bioluminescence imaging that the MSC migration at the fracture site is time- and dose-dependent and, it is exclusively CXCR4-dependent. MSC improved the fracture healing affecting the callus biomechanical properties and such improvement correlated with an increase in cartilage and bone content, and changes in callus morphology as determined by micro-computed tomography and histological studies. Transplanting CMV-Cre-R26R-Lac Z-MSC, we found that MSCs engrafted within the callus endosteal niche. Using MSCs from BMP-2-Lac Z mice genetically modified using a bacterial artificial chromosome system to be beta-gal reporters for bone morphogenic protein 2 (BMP-2) expression, we found that MSCs contributed to the callus initiation by expressing BMP-2. The knowledge of the multiple MSC regenerative abilities in fracture healing will allow design of novel MSC-based therapies to treat fractures. PMID- 19544446 TI - Adult palatum as a novel source of neural crest-related stem cells. AB - Somatic neural and neural crest stem cells are promising sources for cellular therapy of several neurodegenerative diseases. However, because of practical considerations such as inadequate accessibility of the source material, the application of neural crest stem cells is strictly limited. The secondary palate is a highly regenerative and heavily innervated tissue, which develops embryonically under direct contribution of neural crest cells. Here, we describe for the first time the presence of nestin-positive neural crest-related stem cells within Meissner corpuscles and Merkel cell-neurite complexes located in the hard palate of adult Wistar rats. After isolation, palatal neural crest-related stem cells (pNC-SCs) were cultivated in the presence of epidermal growth factor and fibroblast growth factor under serum-free conditions, resulting in large amounts of neurospheres. We used immunocytochemical techniques and reverse transcriptase-polymerase chain reaction to assess the expression profile of pNC SCs. In addition to the expression of neural crest stem cell markers such as Nestin, Sox2, and p75, we detected the expression of Klf4, Oct4, and c-Myc. pNC SCs differentiated efficiently into neuronal and glial cells. Finally, we investigated the potential expression of stemness markers within the human palate. We identified expression of stem cell markers nestin and CD133 and the transcription factors needed for reprogramming of somatic cells into pluripotent cells: Sox2, Oct4, Klf4, and c-Myc. These data show that cells isolated from palatal rugae form neurospheres, are highly plastic, and express neural crest stem cell markers. In addition, pNC-SCs may have the ability to differentiate into functional neurons and glial cells, serving as a starting point for therapeutic studies. PMID- 19544447 TI - Wnt3a-induced mesoderm formation and cardiomyogenesis in human embryonic stem cells. AB - In vitro differentiation of human embryonic stem cells (hESCs) into pure human cardiomyocytes (hESCMs) would present a powerful tool to further the creation of cell models designed to advance preclinical drug development. Here, we report a novel differentiation method to substantially increase hESCM yield. Upon early and transient treatment of hESCs with Wnt3a, embryoid body and mesendoderm formation is enhanced, leading to greater differentiation toward cardiomyocytes. Moreover, the generated beating clusters are highly enriched with cardiomyocytes (50%) and express genes characteristic of cardiac cells, providing evidence that these hESCMs are competent to develop in vitro into functional and physiologically relevant cardiomyocytes. In summary, this protocol not only has the potential to guarantee a renewable supply of enriched cardiomyocyte populations for developing novel and more predictive cell models, but it also should provide valuable insights into pathways critical for cardiac regeneration. PMID- 19544448 TI - Low oxygen enhances primitive and definitive neural stem cell colony formation by inhibiting distinct cell death pathways. AB - Neural stem cells (NSCs) can be derived from single mouse embryonic stem cells (ESCs) in the absence of instructive factors. Clonal primitive NSC (pNSC) colonies are formed first, and then give rise to clonal, fibroblast growth factor dependent definitive neural stem cells (dNSCs). We tested low-oxygen culture as a potential method of alleviating the extensive cell death seen in pNSCs and dNSCs. Culture in low (4%) oxygen promoted survival of pNSCs by inhibiting apoptosis inducing factor (AIF)-dependent cell death, although pNSCs undergo both AIF- and caspase-mediated cell death in 20% oxygen. In contrast, survival of dNSCs in low oxygen was increased by inhibition of caspase-dependent cell death. In normoxia, AIF is implicated in promoting dNSC survival. Neither survival effect was dependent on the main transcriptional effector of hypoxia, hypoxia-inducible factor 1. Low-oxygen concentrations may be involved in expansion of early NSC populations by inhibiting cell death through different pathways in these sequential pNSC and dNSC populations. PMID- 19544449 TI - Monitoring a correctional mental health care system: the role of the mental health expert. AB - Class action litigation has been instrumental in jail and prison reform over the past four decades. This article provides a very brief introduction underlying the legal basis for such litigation. It focuses on the role of the mental health expert in monitoring a correctional mental health care system as a result of class action litigation including issues related to selection of the expert, development of the remedial plan, and monitoring of the implementation of the remedial plan. The importance of policies and procedures and a quality improvement process is emphasized. Essential elements of the monitoring process, prior to and during the site assessment, are described. Inmates and correctional staff alike have benefited substantially from such litigation in the form of increased resources and positive changes in institutional culture. PMID- 19544450 TI - ES cell cycle progression and differentiation require the action of the histone methyltransferase Dot1L. AB - Mouse embryonic stem cells (ESCs) proliferate with rapid cell cycle kinetics but without loss of pluripotency. The histone methyltransferase Dot1L is responsible for methylation of histone H3 at lysine 79 (H3K79me). We investigated whether ESCs require Dot1L for proper stem cell behavior. ESCs deficient in Dot1L tolerate a nearly complete loss of H3K79 methylation without a substantial impact on proliferation or morphology. However, shortly after differentiation is induced, Dot1L-deficient cells cease proliferating and arrest in G2/M-phase of the cell cycle, with increased levels of aneuploidy. In addition, many aberrant mitotic spindles occur in Dot1L-deficient cells. Surprisingly, these mitotic and cell cycle defects fail to trigger apoptosis, indicating that mouse ESCs lack stringent cell cycle checkpoint control during initial stages of differentiation. Transcriptome analysis indicates that Dot1L deficiency causes the misregulation of a select set of genes, including many with known roles in cell cycle control and cellular proliferation as well as markers of endoderm differentiation. The data indicate a requirement for Dot1L function for early stages of ESC differentiation where Dot1L is necessary for faithful execution of mitosis and proper transcription of many genes throughout the genome. PMID- 19544451 TI - Bone marrow mononuclear cells have neurovascular tropism and improve diabetic neuropathy. AB - Bone marrow-derived mononuclear cells (BMNCs) have been shown to effectively treat ischemic cardiovascular diseases. Because diabetic neuropathy (DN) is causally associated with impaired angiogenesis and deficiency of angiogenic and neurotrophic factors in the nerves, we investigated whether DN can be ameliorated by local injection of BMNCs. Severe peripheral neuropathy, characterized by a significant decrease in the motor and sensory nerve conduction velocities (NCVs), developed 12 weeks after the induction of diabetes with streptozotocin in rats. The injection of BMNCs restored motor and sensory NCVs to normal levels and significantly improved vascular density and blood flow in diabetic nerves over 4 weeks. Fluorescent microscopic observation revealed that DiI-labeled BMNCs preferentially engrafted in sciatic nerves. Whole-mount fluorescent imaging and confocal microscopic evaluation demonstrated that many of the BMNCs localized following the course of the vasa nervorum in close proximity to blood vessels without incorporation into vasa nervorum as endothelial cells at a detectable level. Real-time reverse transcription-polymerase chain reaction analysis showed that the levels of angiogenic and neurotrophic factors were significantly increased in the nerves by BMNC injection. Local transplantation of BMNCs improved experimental DN by augmenting angiogenesis and increasing angiogenic and neurotrophic factors in peripheral nerves. These findings suggest that BMNC transplantation may represent a novel therapeutic option for treating DN. PMID- 19544452 TI - Zinc finger protein 191 (ZNF191/Zfp191) is necessary to maintain neural cells as cycling progenitors. AB - The identification of the factors that allow better monitoring of stem cell renewal and differentiation is of paramount importance for the implementation of new regenerative therapies, especially with regard to the nervous and hematopoietic systems. In this article, we present new information on the function of zinc finger protein 191 (ZNF/Zfp191), a factor isolated in hematopoietic cell lines, within progenitors of the central nervous system (CNS). ZNF/Zfp191 has been found to be principally expressed in progenitors of the developing CNS of humans and mice. Such an overlap of the expression patterns in addition to the high homology of the protein in mammals suggested that ZNF/Zfp191 exerts a conserved function within such progenitors. Indeed, ZNF191 knockdown in human neural progenitors inhibits proliferation and leads to the exit of the cell cycle. Conversely, ZNF191 misexpression maintains progenitors in cycle and exerts negative control on the Notch pathway, which prevents them from differentiating. The present data, together with the fact that the inactivation of Zfp191 leads to embryonic lethality, confirm ZNF191 as an essential factor acting for the promotion of the cell cycle and thus maintenance in the progenitor stage. On the bases of expression data, such a function can be extended to progenitor cells of other tissues such as the hematopoietic system, which emphasizes the important issue of further understanding the molecular events controlled by ZNF/Zfp191. PMID- 19544453 TI - DNER, an epigenetically modulated gene, regulates glioblastoma-derived neurosphere cell differentiation and tumor propagation. AB - Neurospheres derived from glioblastoma (GBM) and other solid malignancies contain neoplastic stem-like cells that efficiently propagate tumor growth and resist cytotoxic therapeutics. The primary objective of this study was to use histone modifying agents to elucidate mechanisms by which the phenotype and tumor promoting capacity of GBM-derived neoplastic stem-like cells are regulated. Using established GBM-derived neurosphere lines and low passage primary GBM-derived neurospheres, we show that histone deacetylase (HDAC) inhibitors inhibit growth, induce differentiation, and induce apoptosis of neoplastic neurosphere cells. A specific gene product induced by HDAC inhibition, Delta/Notch-like epidermal growth factor-related receptor (DNER), inhibited the growth of GBM-derived neurospheres, induced their differentiation in vivo and in vitro, and inhibited their engraftment and growth as tumor xenografts. The differentiating and tumor suppressive effects of DNER, a noncanonical Notch ligand, contrast with the previously established tumor-promoting effects of canonical Notch signaling in brain cancer stem-like cells. Our findings are the first to implicate noncanonical Notch signaling in the regulation of neoplastic stem-like cells and suggest novel neoplastic stem cell targeting treatment strategies for GBM and potentially other solid malignancies. PMID- 19544454 TI - HOXA3 modulates injury-induced mobilization and recruitment of bone marrow derived cells. AB - The regulated recruitment and differentiation of multipotent bone marrow-derived cells (BMDCs) to sites of injury are critical for efficient wound healing. Previously we demonstrated that sustained expression of HOXA3 both accelerated wound healing and promoted angiogenesis in diabetic mice. In this study, we have used green fluorescent protein-positive bone marrow chimeras to investigate the effect of HOXA3 expression on recruitment of BMDCs to wounds. We hypothesized that the enhanced neovascularization induced by HOXA3 is due to enhanced mobilization, recruitment, and/or differentiation of BMDCs. Here we show that diabetic mice treated with HOXA3 displayed a significant increase in both mobilization and recruitment of endothelial progenitor cells compared with control mice. Importantly, we also found that HOXA3-treated mice had significantly fewer inflammatory cells recruited to the wound compared with control mice. Microarray analyses of HOXA3-treated wounds revealed that indeed HOXA3 locally increased expression of genes that selectively promote stem/progenitor cell mobilization and recruitment while also suppressing expression of numerous members of the proinflammatory nuclear factor kappaB pathway, including myeloid differentiation primary response gene 88 and toll interacting protein. Thus HOXA3 accelerates wound repair by mobilizing endothelial progenitor cells and attenuating the excessive inflammatory response of chronic wounds. PMID- 19544455 TI - Stem cell therapy restores transparency to defective murine corneas. AB - Corneal scarring from trauma and inflammation disrupts vision for millions worldwide, but corneal transplantation, the primary therapy for corneal blindness, is unavailable to many affected individuals. In this study, stem cells isolated from adult human corneal stroma were examined for the ability to correct stromal opacity in a murine model by direct injection of cells into the corneal stroma. In wild-type mice, injected human stem cells remained viable for months without fusing with host cells or eliciting an immune T-cell response. Human corneal-specific extracellular matrix, including the proteoglycans lumican and keratocan, accumulated in the treated corneas. Lumican-null mice have corneal opacity similar to that of scar tissue as a result of disruption of stromal collagen organization. After injection with human stromal stem cells, stromal thickness and collagen fibril defects in these mice were restored to that of normal mice. Corneal transparency in the treated mice was indistinguishable from that of wild-type mice. These results support the immune privilege of adult stem cells and the ability of stem cell therapy to regenerate tissue in a manner analogous to organogenesis and clearly different from that of normal wound healing. The results suggest that cell-based therapy can be an effective approach to treatment of human corneal blindness. PMID- 19544458 TI - MicroRNA 92b controls the G1/S checkpoint gene p57 in human embryonic stem cells. AB - Human embryonic stem (ES) cells exhibit a shorter G(1) cell cycle phase than most somatic cells. Here, we examine the role of an abundant, human ES cell-enriched microRNA, miR-92b, in cell cycle distribution. Inhibition of miR-92b in human ES cells results in a greater number of cells in the G(1) phase and a lower number in the S phase. Conversely, overexpression of miR-92b in differentiated cells results in a decreased number of cells in G1 phase and an increased number in S phase. p57, a gene whose product inhibits G(1) to S-phase progression, is one of the predicted targets of miR-92b. Inhibition of miR-92b in human ES cells increases p57 protein levels, and miR-92b overexpression in differentiated cells decreases p57 protein levels. Furthermore, miR-92b inhibits a luciferase reporter construct that includes part of the 3' untranslated region of the p57 gene containing the predicted target of the miR-92b seed sequence. Thus, we show that the miRNA miR-92b directly downregulates protein levels of the G(1)/S checkpoint gene p57. STEM CELLS 2009;27:1524-1528. PMID- 19544457 TI - Transplantation of embryonic and adult neural stem cells in the granuloprival cerebellum of the weaver mutant mouse. AB - Numerous studies have explored the potential of different stem and progenitor cells to replace at-risk neuronal populations in a variety of neurodegenerative disease models. This study presents data from a side-by-side approach of engrafting two different stem/progenitor cell populations within the postnatal cerebellum of the weaver neurological mutant mouse--cerebellar-derived multipotent astrocytic stem cells and embryonic stem cell-derived neural precursors--for comparative analysis. We show here that both donor populations survive, migrate, and appear to initiate differentiation into neurons within the granuloprival host environment. Neither of these disparate stem/progenitor cell populations adopted significant region-specific identities, despite earlier studies that suggested the potential of these cells to respond to in vivo cues when placed in a permissive/instructive milieu. However, data presented here suggest that molecular and cellular deficits present within weaver homozygous or heterozygous brains may promote a slightly more positive donor cell response toward acquisition of a neuronal phenotype. Hence, it is likely that a fine balance exists between a compromised host environment that is amenable to cell replacement and that of a degenerating cellular milieu where it is perhaps too deleterious to support extensive neuronal differentiation and functional cellular integration. These findings join a growing list of studies that show successful cell replacement depends largely on the interplay between the potentiality of the donor cells and the specific pathological conditions of the recipient environment, and that emergent therapies for neurological disorders involving the use of neural stem cells still require refinement. PMID- 19544459 TI - Embryonic stem cell-derived cardiomyogenesis: a novel role for calreticulin as a regulator. AB - A role for calreticulin, an endoplasmic reticulum (ER)-resident, Ca(2+)-binding chaperone, has recently emerged in the context of cardiomyogenesis. We previously proposed calreticulin to be a novel cardiac fetal gene, because calreticulin knockout causes embryonic lethality in mice as a result of cardiac defects, it is transiently activated during heart development, and heart-targeted overexpression of constitutively active calcineurin in calreticulin-null mice rescues the lethal phenotype. Calreticulin affects Ca(2+) homeostasis and expression of adhesion related genes. Using cardiomyocytes derived from both calreticulin-null and wild type embryonic stem (ES) cells, we show here that cardiomyogenesis from calreticulin-null ES cells is accelerated but deregulated, such that the myofibrils of calreticulin-null cardiomyocytes become disorganized and disintegrate with time in culture. We have previously shown that the disorganization of the actin cytoskeleton in calreticulin-null cells may be explained, at least in part, by the downregulation of adhesion proteins, implying that calreticulin ablation causes adhesion-related defects. Here, upon examination of adhesion proteins, we found that vinculin is downregulated in calreticulin-null cardiomyocytes. We also found c-Src activity to be higher in calreticulin-null cardiomyocytes than in wild-type cardiomyocytes, and c-Src activity is affected by both calreticulin and [Ca(2+)]. Finally, we show that calreticulin and calsequestrin, the major Ca(2+) storage proteins of the ER and sarcoplasmic reticulum, respectively, exhibit alternate distributions. This suggests that calreticulin may have a housekeeping role to play in mature cardiomyocytes as well as during cardiomyogenesis. We propose here that calreticulin, an ER Ca(2+) storage protein, is a crucial regulator of cardiomyogenesis whose presence is required for controlled cardiomyocyte development from ES cells. PMID- 19544456 TI - Differentiation of a highly tumorigenic basal cell compartment in urothelial carcinoma. AB - Highly tumorigenic cancer cell (HTC) populations have been identified for a variety of solid tumors and assigned stem cell properties. Strategies for identifying HTCs in solid tumors have been primarily empirical rather than rational, particularly in epithelial tumors, which are responsible for 80% of cancer deaths. We report evidence for a spatially restricted bladder epithelial (urothelial) differentiation program in primary urothelial cancers (UCs) and in UC xenografts. We identified a highly tumorigenic UC cell compartment that resembles benign urothelial stem cells (basal cells), co-expresses the 67-kDa laminin receptor and the basal cell-specific cytokeratin CK17, and lacks the carcinoembryonic antigen family member CEACAM6 (CD66c). This multipotent compartment resides at the tumor-stroma interface, is easily identified on histologic sections, and possesses most, if not all, of the engraftable tumor forming ability in the parental xenograft. We analyzed differential expression of genes and pathways in basal-like cells versus more differentiated cells. Among these, we found significant enrichment of pathways comprising "hallmarks" of cancer, and pharmacologically targetable signaling pathways, including Janus kinase-signal transducer and activator of transcription, Notch, focal adhesion, mammalian target of rapamycin, epidermal growth factor receptor (erythroblastic leukemia viral oncogene homolog [ErbB]), and wingless-type MMTV integration site family (Wnt). The basal/HTC gene expression signature was essentially invisible within the context of nontumorigenic cell gene expression and overlapped significantly with genes driving progression and death in primary human UC. The spatially restricted epithelial differentiation program described here represents a conceptual advance in understanding cellular heterogeneity of carcinomas and identifies basal-like HTCs as attractive targets for cancer therapy. PMID- 19544460 TI - Trafficking mesenchymal stem cell engraftment and differentiation in tumor bearing mice by bioluminescence imaging. AB - The objective of the study was to track the distribution and differentiation of mesenchymal stem cells (MSCs) in tumor-bearing mice. The 4T1 murine breast cancer cells were labeled with renilla luciferase-monomeric red fluorescence protein (rLuc-mRFP) reporter gene. The MSCs labeled with firefly luciferase-enhanced green fluorescence protein (fLuc-eGFP) reporter gene (MSCs-R) were isolated from L2G85 transgenic mice that constitutively express fLuc-eGFP reporter gene. To study the tumor tropism of MSCs, we established both subcutaneous and lung metastasis models. In lung metastasis tumor mice, we injected MSCs-R intravenously either on the same day or 4 days after 4T1 tumor cell injection. In subcutaneous tumor mice, we injected MSCs-R intravenously 7 days after subcutaneous 4T1 tumor inoculation. The tumor growth was monitored by rLuc bioluminescence imaging (BLI). The fate of MSCs-R was monitored by fLuc BLI. The localization of MSCs-R in tumors was examined histologically. The osteogenic and adipogenic differentiation of MSCs-R was investigated by alizarin red S and oil red O staining, respectively. The mechanism of the dissimilar differentiation potential of MSCs-R under different tumor microenvironments was investigated. We found that the 4T1 cells were successfully labeled with rLuc-mRFP. The MSCs-R isolated from L2G85 transgenic mice constitutively express fLuc-eGFP reporter gene. When injected intravenously, MSCs-R survived, proliferated, and differentiated in tumor sites but not elsewhere. The localization of GFP(+) MSCs R in tumor lesions was confirmed ex vivo. In conclusion, the MSCs-R can selectively localize, survive, and proliferate in both subcutaneous tumor and lung metastasis as evidenced by noninvasive bioluminescence imaging and ex vivo validation. The MSCs-R migrated to lung tumor differentiated into osteoblasts, whereas the MSCs-R targeting subcutaneous tumor differentiated into adipocytes. PMID- 19544461 TI - Maintenance of undifferentiated state and self-renewal of embryonic neural stem cells by Polycomb protein Ring1B. AB - Cell lineages generated during development and tissue maintenance are derived from self-renewing stem cells by differentiation of their committed progeny. Recent studies suggest that epigenetic mechanisms, and in particular the Polycomb group (PcG) of genes, play important roles in controlling stem cell self-renewal. Here, we address PcG regulation of stem cell self-renewal and differentiation through inactivation of Ring1B, a histone H2A E3 monoubiquitin ligase, in embryonic neural stem cells (NSCs) from the olfactory bulb of a conditional mouse mutant line. We show that neural stem/progenitor cell proliferation in vivo and in neurosphere assays is impaired, lacking Ring1B, and their self-renewal and multipotential abilities, assessed as sphere formation and differentiation from single cells, are severely affected. We also observed unscheduled neuronal, but not glial, differentiation of mutant stem/progenitor cells under proliferating conditions, an alteration enhanced in cells also lacking Ring1A, the Ring1B paralog, some of which turned into morphologically identifiable neurons. mRNA analysis of mutant cells showed upregulation of some neuronal differentiation related transcription factors and the cell proliferation inhibitor Cdkn1a/p21, as well as downregulation of effectors of the Notch signaling pathway, a known inhibitor of neuronal differentiation of stem/progenitor cells. In addition, differentiation studies of Ring1B-deficient progenitors showed decreased oligodendrocyte formation in vitro and enhanced neurogenesis and reduced gliogenesis in vivo. These data suggest a role for Ring1B in maintenance of the undifferentiated state of embryonic neural stem/progenitor cells. They also suggest that Ring1B may modulate the differentiation potential of NSCs to neurons and glia. PMID- 19544462 TI - Restoration of Runx1 expression in the Tie2 cell compartment rescues definitive hematopoietic stem cells and extends life of Runx1 knockout animals until birth. AB - Mice deficient in the runt homology domain transcription factor Runx1/AML1 fail to generate functional clonogenic hematopoietic cells and die in utero by embryonic day 12.5. We previously generated Runx1 reversible knockout mice, in which the Runx1 locus can be restored by Cre-mediated recombination. We show here that selective restoration of the Runx1 locus in the Tie2 cell compartment rescues clonogenic hematopoietic progenitors in early Runx1-null embryos and rescues lymphoid and myeloid lineages during fetal development. Furthermore, fetal liver cells isolated from reactivated Runx1 embryos are capable of long term multilineage lymphomyeloid reconstitution of adult irradiated recipients, demonstrating the rescue of definitive hematopoietic stem cells. However, this rescue of the definitive hematopoietic hierarchy is not sufficient to rescue the viability of animals beyond birth, pointing to an essential role for Runx1 in other vital developmental processes. PMID- 19544463 TI - Murine "cardiospheres" are not a source of stem cells with cardiomyogenic potential. AB - Recent remarkable studies have reported that clonogenic putative cardiac stem cells (CSCs) with cardiomyogenic potential migrate from heart tissue biopsies during ex vivo culture, and that these CSCs self-organize into spontaneously beating cardiospheres (CSs). Such data have provided clear promise that injured heart tissue may be repaired by stem cell therapy using autologous CS-derived cells. By further examining CSs from the original CS protocol using immunofluorescence, quantitative reverse transcription-polymerase chain reaction, and microscopic analysis, we here report a more mundane result: that spontaneously beating CSs from neonatal rats likely consist of contaminating myocardial tissue fragments. Thus, filtering away these tissue fragments resulted in CSs without cardiomyogenic potential. Similar data were obtained with CSs derived from neonatal mice as wells as adult rats/mice. Additionally, using in vitro culture, fluorescence-activated cell sorting, and immunofluorescence, we demonstrate that these CSs are generated by cellular aggregation of GATA 4(+)/collagen I(+)/alpha-smooth muscle actin (SMA)(+)/CD45(-) cells rather than by clonal cell growth. In contrast, we found that the previously proposed CS forming cells, dubbed phase bright cells, were GATA-4(-)/collagen I(-)/alpha-SMA( )/CD45(+) and unable to form CSs by themselves. Phenotypically, the CS cells largely resembled fibroblasts, and they lacked cardiomyogenic as well as endothelial differentiation potential. Our data imply that the murine CS model is unsuitable as a source of CSCs with cardiomyogenic potential, a result that is in contrast to previously published data. We therefore suggest, that human CSs should be further characterized with respect to phenotype and differentiation potential before initiating human trials. PMID- 19544465 TI - Bioluminescence imaging of Olig2-neural stem cells reveals improved engraftment in a demyelination mouse model. AB - A major issue in the potential application of neural stem cell (NSC)-based cell replacement therapy for demyelinating diseases is the question of the survival, functional behavior, and stability of implanted NSC-derived oligodendrocyte precursor cells (OPCs) over an extended period. To address this issue, we employed bioluminescence imaging (BLI) as a noninvasive longitudinal in vivo monitoring technique and followed the fate of NSCs isolated from luciferase-green fluorescent protein-actin transgenic mice after stereotactic implantation in the demyelinated corpus callosum of cuprizone-fed mice. We compared normal NSCs with NSCs that were primed to become OPCs by the induction of Olig2 overexpression (Olig2-NSCs). BLI, validated by immunohistochemistry, revealed that, after a steep cell loss after implantation during the first 3 weeks, approximately 10% of the Olig2-NSCs stably survived for 2 months after implantation, in contrast to <1% of the normal NSCs. Immunohistochemistry, at the light and electron microscopic levels, revealed that the majority of the surviving Olig2-NSCs had differentiated into an oligodendrocytic cell lineage and contributed to remyelination of axons in the corpus callosum. The number of axons remyelinated by the implanted cells, however, was a small fraction of the total number of axons remyelinated by endogenous oligodendrocytes. Apparently, most of the implanted NSCs did not survive the transition into an inappropriate non neurogenic niche, compressed by surrounding host tissue, in hostile, inflammatory conditions created by activated microglia. Only the ones that managed to differentiate rapidly into a mature neural cell type and become functionally integrated survived. PMID- 19544464 TI - The role of PTIP in maintaining embryonic stem cell pluripotency. AB - Pax transactivation domain-interacting protein (PTIP) is a ubiquitously expressed, nuclear protein that is part of a histone H3K4 methyltransferase complex and is essential for embryonic development. Methylation of H3K4 is an epigenetic mark found on many critical developmental regulatory genes in embryonic stem (ES) cells and, together with H3K27 methylation, constitutes a bivalent epigenetic signature. To address the function of PTIP in ES cells, we generated ES cell lines from a floxed ptip allele and deleted PTIP function with Cre recombinase. The ptip(-/-) ES cell lines exhibited a high degree of spontaneous differentiation to trophectoderm and a loss of pluripotency. Reduced levels of Oct4 expression and H3K4 methylation were observed. Upon differentiation, ptip(-/-) embryoid bodies showed reduced levels of marker gene expression for all three primary germ layers. These results suggest that the maintenance of H3K4 methylation is essential and requires PTIP function during the in vitro propagation of pluripotent ES cells. PMID- 19544466 TI - Gene targeting in a HUES line of human embryonic stem cells via electroporation. AB - Genetic modification is critical for achieving the full potential of human embryonic stem (ES) cells as a tool for therapeutic development and for basic research. Targeted modifications in human ES cells have met with limited success because of the unique culture conditions for many human ES cell lines. The HUES lines of human ES cells were developed for ease of manipulation and are gaining increased utility in stem cell research. We tested conditions for gene targeting via electroporation in the HUES-9 human ES cell line and demonstrate here successful gene targeting at the gene encoding Fezf2 (also known as Fezl), a transcription factor involved in corticospinal neuron development. With a targeting strategy involving positive and negative selection that is applicable to all genes, we observed a gene targeting frequency of approximately 1.5% for Fezf2, a gene not expressed in human ES cells. We found that conditions developed for gene targeting in mouse ES cells can be readily adapted to HUES cells with few key modifications. HUES-9 cells exhibit an intrinsically high efficiency of clonal expansion and sustain electroporation-based gene targeting procedures without any significant loss of pluripotency marker expression or karyotypic stability. Thus, human ES cell lines adapted for enzymatic passage and efficient clonal expansion can be highly amenable to genetic modifications, which will facilitate their application in basic science and clinical development. PMID- 19544467 TI - Embryonic stem cells and mammary luminal progenitors directly sense and respond to microbial products. AB - Stem cells are normally maintained in a quiescent state and proliferate only under certain conditions; however, little is known about the biological stimuli that initiate the proliferation and differentiation of stem cells. In this study, we found that functional Toll-like receptors (TLRs) are expressed on mouse embryonic stem (ES) cells and that TLR ligands stimulate ES cell proliferation and promote their hematopoietic differentiation. TLR ligands activate TLR mediated signaling pathways, leading to the altered expression of numerous genes in ES cells. Moreover, TLR ligands efficiently stimulate the proliferation and expansion of adult stem cells and progenitors of nonhematopoietic tissues, such as mammary glands and intestine as well. We further found that mammary luminal progenitor cells (Lin(-)CD29(+)CD61(+)) express TLR4-MD2 complex and actively proliferate, resulting in the enhanced growth of mammospheres in response to TLR ligands. Thus, mouse ES cells and adult tissue-specific stem cells/progenitors directly sense and respond to microbial products, which function as a class of foreign, but biological stimuli for stem cell/progenitor proliferation. This finding expands the biological role of TLRs and has implications in understanding stem cell biology, tissue repair/homeostasis, and the role of infection and inflammation in malignant transformation. PMID- 19544468 TI - Regulation of boundary cap neural crest stem cell differentiation after transplantation. AB - Success of cell replacement therapies for neurological disorders will depend largely on the optimization of strategies to enhance viability and control the developmental fate of stem cells after transplantation. Once transplanted, stem/progenitor cells display a tendency to maintain an undifferentiated phenotype or differentiate into inappropriate cell types. Gain and loss of function experiments have revealed key transcription factors which drive differentiation of immature stem/progenitor cells toward more mature stages and eventually to full differentiation. An attractive course of action to promote survival and direct the differentiation of transplanted stem cells to a specific cell type would therefore be to force expression of regulatory differentiation molecules in already transplanted stem cells, using inducible gene expression systems which can be controlled from the outside. Here, we explore this hypothesis by employing a tetracycline gene regulating system (Tet-On) to drive the differentiation of boundary cap neural crest stem cells (bNCSCs) toward a sensory neuron fate after transplantation. We induced the expression of the key transcription factor Runx1 in Sox10-expressing bNCSCs. Forced expression of Runx1 strongly increased transplant survival in the enriched neurotrophic environment of the dorsal root ganglion cavity, and was sufficient to guide differentiation of bNCSCs toward a nonpeptidergic nociceptive sensory neuron phenotype both in vitro and in vivo after transplantation. These findings suggest that exogenous activation of transcription factors expression after transplantation in stem/progenitor cell grafts can be a constructive approach to control their survival as well as their differentiation to the desired type of cell and that the Tet-system is a useful tool to achieve this. PMID- 19544469 TI - Initiation of dopaminergic differentiation of Nurr1(-) mesencephalic precursor cells depends on activation of multiple mitogen-activated protein kinase pathways. AB - Interleukin-1 (IL-1) plays a pivotal role in terminal dopaminergic differentiation of midbrain-derived neural precursor cells already committed to the mesencephalic dopaminergic phenotype (named mdNPCs for mesencephalic dopaminergic neural precursor cells). Here we characterized the molecular events in long-term expanded rat nuclear receptor related-1(-) (Nurr1(-)) mdNPCs in response to IL-1beta during their terminal dopaminergic specification. We showed that IL-1beta induced a rapid induction of mRNA of dopaminergic key fate determining transcription factors, such as Nurr1 and Pitx3, and a subsequent increase of tyrosine hydroxylase protein as an early marker for dopaminergic neurons in vitro. These effects of IL-1beta were specific for mdNPCs and were not observed in striatal neural precursor cells (NPCs). Surprisingly, IL-1beta did not activate the NF-kappaB pathway or the transcription factor activating protein 1 (AP-1), but inhibition of nuclear translocation of NF-kappaB by SN50 facilitated IL-1beta-induced Nurr1 expression and dopaminergic differentiation of mdNPCs. Incubation of mdNPCs with IL-1beta led to a rapid phosphorylation of ERK1/2 and p38 mitogen-activated protein (MAP) kinases within 1 to 3 hours, whereas Jun kinase was not phosphorylated in response to IL-1beta. Consistently, inhibition of the ERK1/2 pathway or p38 MAP kinase blocked Nurr1 upregulation and further dopaminergic specification of mdNPCs, but not differentiation into MAP2ab(+) neurons. IL-1 receptor antagonist did not block early dopaminergic differentiation events, suggesting that the effects of IL-1beta are not mediated through activation of IL-1 receptor type I. Our results indicate that induction of terminal dopaminergic specification of Nurr1(-) mdNPCs by IL-1beta depends on activation of the ERK1/2 and p38 MAP kinase pathway. PMID- 19544470 TI - p38 MAP kinase inhibits neutrophil development through phosphorylation of C/EBPalpha on serine 21. AB - Many extracellular stimuli regulate growth, survival, and differentiation responses through activation of the dual specificity mitogen activated protein kinase (MAPK) kinase three (MKK3) and its downstream effector p38 MAPK. Using CD34+ hematopoietic progenitor cells, here we describe a novel role for MKK3 p38MAPK in the regulation of myelopoiesis. Inhibition of p38MAPK utilizing the pharmacological inhibitor SB203580, enhanced neutrophil development ex vivo, but conversely reduced eosinophil differentiation. In contrast, constitutive activation of MKK3 dramatically inhibited neutrophil differentiation. Transplantation of beta2-microglobulin(-/-) nonobese diabetic/severe combined immune deficient (NOD/SCID) mice with CD34+ cells ectopically expressing constitutively active MKK3 resulted in reduced neutrophil differentiation in vivo, whereas eosinophil development was enhanced. Inhibitory phosphorylation of CCAAT/enhancer binding protein alpha (C/EBPalpha) on serine 21 was induced upon activation of p38MAPK. Moreover, ectopic expression of a non-phosphorylatable C/EBPalpha mutant was sufficient to abrogate MKK3-induced inhibition of neutrophil development. Furthermore, treatment of CD34+ progenitors from patients with severe congenital neutropenia with SB203580 restored neutrophil development. These results establish a novel role for MKK3-p38MAPK in the regulation of lineage choices during myelopoiesis through modulation of C/EBPalpha activity. This signaling module may thus provide an important therapeutic target in the treatment of bone marrow failure. PMID- 19544471 TI - Evaluating erythropoietin-associated tumor progression using archival tissues from a phase III clinical trial. AB - Despite the prevalence of anemia in cancer, recombinant erythropoietin (Epo) has declined in use because of recent Phase III trials showing more rapid cancer progression and reduced survival in subjects randomized to Epo. Since Epo receptor (EpoR), Jak2, and Hsp70 are well-characterized mediators of Epo signaling in erythroid cells, we hypothesized that Epo might be especially harmful in patients whose tumors express high levels of these effectors. Because of the insensitivity of immunohistochemistry for detecting low level EpoR protein, we developed assays to measure levels of EpoR, Jak2 and Hsp70 mRNA in formalin-fixed paraffin-embedded (FFPE) tumors. We tested 23 archival breast tumors as well as 136 archival head and neck cancers from ENHANCE, a Phase III trial of 351 patients randomized to Epo versus placebo concomitant with radiotherapy following complete resection, partial resection, or no resection of tumor. EpoR, Jak2, and Hsp70 mRNA levels varied >30-fold, >12-fold, and >13-fold across the breast cancers, and >30-fold, >40-fold, and >30-fold across the head and neck cancers, respectively. Locoregional progression-free survival (LPFS) did not differ among patients whose head and neck cancers expressed above- versus below-median levels of EpoR, Jak2 or Hsp70, except in the subgroup of patients with unresected tumors (n = 28), where above-median EpoR, above-median Jak2, and below-median Hsp70 mRNA levels were all associated with significantly poorer LPFS. Our results provide a framework for exploring the relationship between Epo, cancer progression, and survival using archival tumors from other Phase III clinical trials. PMID- 19544472 TI - Mesenchymal stem cells pretreated with delivered Hph-1-Hsp70 protein are protected from hypoxia-mediated cell death and rescue heart functions from myocardial injury. AB - Mesenchymal stem cell (MSC) therapy for myocardial injury has inherent limitations due to the poor viability of MSCs after cell transplantation. In this study, we directly delivered Hsp70, a protein with protective functions against stress, into MSCs, using the Hph-1 protein transduction domain ex vivo for high transfection efficiency and low cytotoxicity. Compared to control MSCs in in vitro hypoxic conditions, MSCs delivered with Hph-1-Hsp70 (Hph-1-Hsp70-MSCs) displayed higher viability and anti-apoptotic properties, including Bcl2 increase, reduction of Bax, JNK phosphorylation and caspase-3 activity. Hsp70 delivery also attenuated cellular ATP-depleting stress. Eight animals per group were used for in vivo experiments after occlusion of the left coronary artery. Transplantation of Hph-1-Hsp70-MSCs led to a decrease in the fibrotic heart area, and significantly reduced the apoptotic positive index by 19.5 +/- 2%, compared to no-treatment controls. Hph-1-Hsp70-MSCs were well-integrated into the infarcted host myocardium. The mean microvessel count per field in the infarcted myocardium of the Hph-1-Hsp70-MSC-treated group (122.1 +/- 13.5) increased relative to the MSC-treated group (75.9 +/- 10.4). By echocardiography, transplantation of Hph-1-Hsp70-MSCs resulted in additional increases in heart function, compared to the MSCs-transplanted group. Our results may help formulate better clinical strategies for in vivo MSC cell therapy for myocardial damage. PMID- 19544473 TI - Snail and slug mediate radioresistance and chemoresistance by antagonizing p53 mediated apoptosis and acquiring a stem-like phenotype in ovarian cancer cells. AB - The transcriptional repressors Snail and Slug contribute to cancer progression by mediating epithelial-mesenchymal transition (EMT), which results in tumor cell invasion and metastases. We extend this current understanding to demonstrate their involvement in the development of resistance to radiation and paclitaxel. The process is orchestrated through the acquisition of a novel subset of gene targets that is repressed under conditions of stress, effectively inactivating p53-mediated apoptosis, while another subset of targets continues to mediate EMT. Repressive activities are complemented by a concurrent derepression of specific genes resulting in the acquisition of stem cell-like characteristics. Such cells are bestowed with three critical capabilities, namely EMT, resistance to p53 mediated apoptosis, and a self-renewal program, that together define the functionality and survival of metastatic cancer stem cells. EMT provides a mechanism of escape to a new, less adverse niche; resistance to apoptosis ensures cell survival in conditions of stress in the primary tumor; whereas acquisition of "stemness" ensures generation of the critical tumor mass required for progression of micrometastases to macrometastases. Our findings, besides achieving considerable expansion of the inventory of direct genes targets, more importantly demonstrate that such elegant cooperative modulation of gene regulation mediated by Snail and Slug is critical for a cancer cell to acquire stem cell characteristics toward resisting radiotherapy- or chemotherapy-mediated cellular stress, and this may be a determinative aspect of aggressive cancer metastases. PMID- 19544474 TI - Astrocytes reverted to a neural progenitor-like state with transforming growth factor alpha are sensitized to cancerous transformation. AB - Gliomas, the most frequent primitive central nervous system tumors, have been suggested to originate from astrocytes or from neural progenitors/stem cells. However, the precise identity of the cells at the origin of gliomas remains a matter of debate because no pre-neoplastic state has been yet identified. Transforming growth factor (TGF)-alpha, an epidermal growth factor family member, is frequently overexpressed in the early stages of glioma progression. We previously demonstrated that prolonged exposure of astrocytes to TGF-alpha is sufficient to trigger their reversion to a neural progenitor-like state. To determine whether TGF-alpha dedifferentiating effects are associated with cancerous transforming effects, we grafted intracerebrally dedifferentiated astrocytes. We show that these cells had the same cytogenomic profile as astrocytes, survived in vivo, and did not give birth to tumors. When astrocytes dedifferentiated with TGF-alpha were submitted to oncogenic stress using gamma irradiation, they acquired cancerous properties: they were immortalized, showed cytogenomic abnormalities, and formed high-grade glioma-like tumors after brain grafting. In contrast, irradiation did not modify the lifespan of astrocytes cultivated in serum-free medium. Addition of TGF-alpha after irradiation did not promote their transformation but decreased their lifespan. These results demonstrate that reversion of mature astrocytes to an embryonic state without genomic manipulation is sufficient to sensitize them to oncogenic stress. PMID- 19544476 TI - Technical aspects and clinical applications of measuring BCR-ABL1 transcripts number in chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is a myeloproliferative disorder characterized by a triphasic clinical course, the morphologic expansion of a terminally differentiated myeloid cell and the presence of the BCR-ABL1 fusion gene, the hallmark of CML. The fusion gene is usually, but not always, associated with a Philadelphia chromosome, the result of a reciprocal exchange of genetic material between chromosome 22 and chromosome 9, which leads to the production of the activated BCR-ABL1 gene and oncoprotein. The breakpoint in the BCR gene occurs commonly downstream of exons e13 or e14 (M-BCR) and less frequently downstream of exons e1 and e2 (m-BCR). Less than 1% of cases carry a breakpoint downstream of exon 6 or 8 ("variant fusion genes") or exon 19 (mu-BCR). Breakpoints in the ABL1 gene cluster upstream of exon a2 (or of exon a3 in less than 5% of patients with CML). Conventional cytogenetic, fluorescence in situ hybridization, and molecular testing for the BCR-ABL1 fusion gene are key investigations for the diagnosis and monitoring of CML. Treatment using tyrosine kinase inhibitors has revolutionized the management of CML with hematologic and cytogenetic response within 12-18 months observed in >85% of patients. Nevertheless, between 15 and 20% of patients may evolve to blastic phase. Measurement of low level or "minimal" residual disease using molecular tests is becoming the gold-standard approach to measure response to therapy due to its higher sensitivity compared to other routine techniques. The technical aspects and clinical applications of molecular monitoring will be the main focus of this article. PMID- 19544475 TI - Changes in red cell ion transport, reduced intratumoral neovascularization, and some mild motor function abnormalities accompany targeted disruption of the Mouse Kell gene (Kel). AB - Kell (ECE-3), a highly polymorphic blood group glycoprotein, displays more than 30 antigens that produce allo-antibodies and, on red blood cells (RBCs), is complexed through a single disulfide bond with the integral membrane protein, XK. XK is a putative membrane transporter whose absence results in a late onset form of neuromuscular abnormalities known as the McLeod syndrome. Although Kell glycoprotein is known to be an endothelin-3-converting enzyme, the full extent of its physiological function is unknown. To study the functions of Kell glycoprotein, we undertook targeted disruption of the murine Kel gene by homologous recombination. RBCs from Kel(-/-) mice lacked Kell glycoprotein, Kell/XK complex, and endothelin-3-converting enzyme activity and had reduced levels of XK. XK mRNA levels in spleen, brain, and testis were unchanged. In Kel( /-) mice RBC Gardos channel activity was increased and the normal enhancement by endothelin-3 was blunted. Analysis of the microvessels of tumors produced from LL2 cells indicated that the central portion of tumors from wild-type mice were populated with many mature blood vessels, but that vessels in tumors from Kel(-/ ) mice were fewer and smaller. The absence of Kell glycoprotein mildly affected some motor activities identified by foot splay on the drop tests. The targeted disruption of Kel in mouse enabled us to identify phenotypes that would not be easily detected in humans lacking Kell glycoprotein. In this regard, the Kell knockout mouse provides a good animal model for the study of normal and/or pathophysiological functions of Kell glycoprotein. PMID- 19544477 TI - Paraneoplastic pemphigus associated with systemic mastocytosis. PMID- 19544478 TI - Synthesis, antibacterial and antifungal activities of novel 1,2,4-triazolium derivatives. AB - A series of novel 1,2,4-triazolium derivatives was synthesized starting from commercially available 1H-1,2,4-triazole, 2,4-dichlorobenzyl chloride, or 2,4 difluorobenzyl bromide. Their antibacterial and antifungal activities were evaluated against Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Bacillus proteus, Bacillus subtilis, Pseudomonas aeruginosa, Candida albicans ATCC 76615, and Aspergillus fumigatus. All structures of the new compounds were confirmed by NMR, IR, and MS spectra, and elemental analyses. The antimicrobial tests showed that most of synthesized triazolium derivatives exhibit significant antibacterial and antifungal activities in vitro. 1-(2,4-Difluorobenzyl)-4 dodecyl-1H-1,2,4-triazol-4-ium bromide and 1-(2,4-Dichlorobenzyl)-4-dodecyl-1H 1,2,4- triazol-4-ium bromide were the most potent compounds against all tested strains with the MIC values ranging from 1.05 to 8.38 microM. They exhibited much stronger activities than the standard drugs chloramphenicol and fluconazole which are in clinical use. The results also showed that the antimicrobial activities of triazolium derivatives depend upon the type of substituent, the length of the alkyl chain, and the number of triazolium rings. PMID- 19544480 TI - Synthesis and antibacterial activities of eperezolid analogs with glycinyl substitutions. AB - A series of eperezolid analogs with glycinyl substitutions were prepared and their antibacterial activities were studied against a panel of susceptible and resistant Gram-positive bacteria. The compounds with N-arylacyl or N heteroarylacyl glycinyl structural units showed good antibacterial activities. The compounds 11b, 11c, and 11e were twofold more active than linezolid against Staphylococcus epidermidis and Enterococcus faecalis. Several pyridine analogs were also prepared and found to have poor antibacterial activity against most of the tested Gram-positive bacteria, however, one of the compounds 12e showed very high activity against Enterococcus faecalis. PMID- 19544479 TI - Design, synthesis, and biological evaluation of prenylated chalcones as vasorelaxant agents. AB - Five prenylated chalcones and one allylated chalcone were prepared according to the analysis based on support vector machine (SVM) classification model. Most of the synthesized chalcones showed potent vasorelaxant activities through evaluation in aortic rings with the endothelium pre-contracted by phenylephrine (PE), indicating that the experimental activities were in good agreement with the theoretical ones. Structure-activity relationship of these compounds showed that the substituent pattern and number of hydroxyl groups were crucial for their vasorelaxant activities and that the replacement of prenyl group with allyl group retained the potent activity. PMID- 19544482 TI - Macroporous photocrosslinked elastomer scaffolds containing microposity: preparation and in vitro degradation properties. AB - The engineering of soft tissue would benefit from the development of effective biodegradable scaffolds capable of dynamic, elastic loading. For this purpose, highly porous, elastomeric scaffolds containing microporous struts were prepared using a dual porogen approach and a photocrosslinkable elastomer. The combination of paraffin microbeads distributed through a water-in-[star-poly(lactide-co epsilon-caprolactone) triacrylate dissolved in ethyl acetate] emulsion followed by photocrosslinking generated a macroporous foam scaffold of average porosities between 90% to 93%, with an average pore diameter of 104 +/- 31 microm with struts containing micropores of 3.1 +/- 2 microm average diameter. The mechanical properties of the scaffolds were readily manipulatable by altering the molecular weight of the star-poly(lactide-co-epsilon-caprolactone) triacrylate prepolymer used. The elastomer scaffolds degraded at the same rate as nonporous polymer samples of the same molecular weight, and exhibited similar changes in mass loss, mechanical properties, and sol fraction during in vitro degradation as found with the nonporous scaffolds. The modulus and stress at break of the scaffolds decreased continuously during degradation while the strain at break remained constant. These scaffolds show potential for use in the engineering of soft tissues. PMID- 19544481 TI - De novo generation of antimicrobial LK peptides with a single tryptophan at the critical amphipathic interface. AB - De novo design of amphipathic model peptides has been successful for generating many antimicrobial peptides with various lengths and amino acid compositions. Here, we suggest a very simple strategy to design antimicrobial peptides with a short length and a simple amino acid composition. Amphipathic helical properties were conferred by using only leucines and lysines and a single tryptophan was positioned at the critical amphipathic interface between the hydrophilic ending side and the hydrophobic starting side, in the helical wheel projection. According to this rule, the model peptides with 7 to 13 residues exhibited antimicrobial activity. Among them, the most potent activity against both Gram positive and Gram-negative bacteria, covering all of the nine bacterial strains tested in this study, was found for the 11-mer sequences having a 1:1 (L(5)K(5)W(6)) or a 3:2 (L(6)K(4)W(6)) ratio of leucines to lysines. In particular, the former peptide L(5)K(5)W(6) could be evaluated as the most useful agent, as it showed no significant hemolytic activity with a broad-spectrum of antimicrobial activity. PMID- 19544483 TI - Contribution of the trifluoroacetyl group in the thermodynamics of antigen antibody binding. AB - We analyzed the binding of the 7C8 antibody to the chloramphenicol phosphonate antigens-one containing a trifluoroacetyl group (CP-F) and the other containing an acetyl group (CP-H)-by using isothermal titration calorimetry (ITC). The thermodynamic difference due to the substitution of F by H was evaluated using free energy calculations based on molecular dynamics (MD) simulations. We have previously shown that another antibody, namely, 6D9, binds more weakly to CP-H than to CP-F, mainly due to the different hydration free energies of the dissociated state and not due to the unfavorable hydrophobic interactions with the antibody in the bound state. Unlike in the binding of the trifluoroacetyl group with 6D9, in its binding with 7C8, it is exposed to the solvent, as seen in the crystal structure of the complex of 7C8 with CP-F. The thermodynamic analysis performed in this study showed that the binding affinity of 7C8 for CP-H is similar to that for CP-F, but this binding to CP-H is accompanied with less favorable enthalpy and more favorable entropy changes. The free energy calculations indicated that, upon the substitution of F by H, enthalpy and entropy changes in the associated and dissociated states were decreased, but the magnitude of enthalpy and entropy changes in the dissociated state was larger than that in the associated state. The differences in binding free energy, enthalpy, and entropy changes determined by the free energy calculations for the substitution of F by H are in good agreement with the experimental results. PMID- 19544484 TI - Evaluation of the effect of intravenous volume expanders upon the volume of distribution of gentamicin in septic neonates. AB - Sepsis has been reported to increase the volume of distribution of gentamicin in neonates. To determine whether this was caused by the use of intravenous volume expanders a retrospective nested case-control study was performed comparing confirmed septic neonates with non-septic controls. Data were collected on intravenous administration of 0.45% saline/10% dextrose, 0.45% saline/5% dextrose, 0.9% normal saline, red blood cells, platelets, immunoglobulin (Intragam P) and albumin. A population pharmacokinetic analysis was performed using NONMEM for 116 neonates (29 confirmed septic) from which 363 gentamicin serum concentrations were available. The final covariate model was CL=0.097x(current weight/2)(1.3)x(postnatal age/7)(0.29) and V=1.07x(current weight/2)(0.8)+(confirmed sepsis)x0.13. The gentamicin volume of distribution was not significantly influenced by the cumulative intravenous volume expanders. The principal covariates that affected the gentamicin volume of distribution were current body weight and confirmed sepsis. PMID- 19544485 TI - Transcriptional and inflammasome-mediated pathways for the induction of IL-1beta production by Mycobacterium tuberculosis. AB - Proinflammatory cytokines of the IL-1 family play an important role for the anti mycobacterial host defense mechanisms. In the present study we have deciphered the pathways leading from recognition of Mycobacterium tuberculosis to the production and release of IL-1beta, the most important member of the IL-1 family. By stimulating cells defective in various pattern recognition receptors, we could demonstrate that IL-1beta production is induced by M. tuberculosis through pathways involving TLR2/TLR6 and NOD2 receptors. In contrast, TLR4, TLR9 and TLR1 receptors are not involved in IL-1beta induction. Recognition of M. tuberculosis by TLR and NOD2 leads to transcription of proIL-1beta through mechanisms involving ERK, p38 and Rip2, but not JNK. Interestingly, although caspase-1 is necessary for the processing of proIL-1beta, activation of caspase-1 is not dependent on the stimulation of cells by M. tuberculosis. Monocytes expressed constitutively active caspase-1. The secretion of IL-1beta is dependent on the activation of P2X7-induced pathways by endogenously released ATP. In conclusion, we have dissected the molecular mechanisms responsible for IL-1beta production by M. tuberculosis, and that may contribute to a deeper knowledge of the mechanisms of cell activation by M. tuberculosis. PMID- 19544486 TI - DC-induced CD8(+) T-cell response is inhibited by MHC class II-dependent DX5(+)CD4(+) Treg. AB - CD4(+) T cells are important for CD8(+) T-cell priming by providing cognate signals for DC maturation. We analyzed the capacity of CD4(+) T cells to influence CD8(+) T-cell responses induced by activated DC. Surprisingly, mice depleted for CD4(+) cells were able to generate stronger antigen-specific CD8(+) T-cell responses after DC vaccination than non-depleted mice. The same observation was made when mice were vaccinated with MHC class II(-/-) DC, indicating the presence of a MHC class II-dependent CD4(+) T-cell population inhibiting CD8(+) T-cell responses. Recently we described the expansion of DX5(+)CD4(+) T cells, a T-cell population displaying immune regulatory properties, upon vaccination with DC. Intriguingly, we now observe an inverse correlation between CD8(+) T-cell induction and expansion of DX5(+)CD4(+) T cells as the latter cells did not expand after vaccination with MHC class II(-/-) DC. In vitro, DX5(+)CD4(+) T cells were able to limit proliferation, modulate cytokine production and induce Foxp3(+) expression in OVA-specific CD8(+) T cells. Together, our data show an inhibitory role of CD4(+) T cells on the induction of CD8(+) T-cell responses by activated DC and indicate the involvement of DX5(+)CD4(+), but not CD4(+)CD25(+), T cells in this process. PMID- 19544489 TI - Mixed-substrate (glycerol tributyrate and fibrin) zymography for simultaneous detection of lipolytic and proteolytic enzymes on a single gel. AB - A new zymography method for simultaneous detection of two different enzymatic activities (lipolytic and proteolytic) using a single SDS-containing or native conformation gel and a mixed-substrate (glycerol tributyrate and fibrin) (MS)(1) gel was developed. After routine electrophoresis, SDS in the gel was removed by treatment with Triton X-100. Gel proteins were electrotransferred to the MS gel. To visualize lipolytic activity, the MS gel was incubated at 37 degrees C (for 6 or 24 h) until clear bands against an opaque background were observed. To detect proteolytic activity, the same MS gel was stained with Coomassie brilliant blue. Using this method, we show that six lipolytic enzymes from Staphylococcus pasteuri NJ-1 and four proteolytic enzymes from two Bacillus strains, B. licheniformis DJ-2 and B. licheniformis NJ-5, isolated from soil, can be simultaneously detected. PMID- 19544488 TI - B7-H3 is a potent inhibitor of human T-cell activation: No evidence for B7-H3 and TREML2 interaction. AB - B7-H3 belongs to the B7 superfamily, a group of molecules that costimulate or down-modulate T-cell responses. Although it was shown that B7-H3 could inhibit T cell responses, several studies - most of them performed in murine systems - found B7-H3 to act in a costimulatory manner. In this study, we have specifically addressed a potential functional dualism of human B7-H3 by assessing the effect of this molecule under varying experimental conditions as well as on different T cell subsets. We show that B7-H3 does not costimulate human T cells. In the presence of strong activating signals, B7-H3 potently and consistently down modulated human T-cell responses. This inhibitory effect was evident when analysing proliferation and cytokine production and affected naive as well as pre activated T cells. Furthermore, we demonstrate that B7-H3-T-cell interaction is characterised by an early suppression of IL-2 and that T-cell inhibition can be reverted by exogenous IL-2. Since the triggering receptor expressed on myeloid cells like transcript 2 (TREML2/TLT-2) has been recently described as costimulatory receptor of murine B7-H3 we have extensively analysed interaction of human B7-H3 with TREML2/TLT-2. In these experiments we found no evidence for such an interaction. Furthermore, our data do not point to a role for murine TREML2 as a receptor for murine B7-H3. PMID- 19544490 TI - Scaling of nucleic acid assays on microelectrophoresis array devices: high dynamic range multi-gene readout from less than ten transcripts. AB - In this paper we describe progress in using the prodigious data-collecting ability of multilane microelectrophoresis instruments to bear on problems in scaled nucleic acid assays. We emphasize compound stacking and solid-support loading as means to concentrate <100 pg samples for direct injection. Reaction Mapping is applied to readout quantitative polymerase chain reaction gene expression and as a way to practically overcome difficulty in interpreting amplification curves of multiplexed quantitative polymerase chain reaction at 20 50 gene/well complexity. We demonstrate multiplexed readout of gene expression over an abundancy range of 9 Log 2 units starting with reverse-transcribed samples as small as five molecules in each sample. PMID- 19544487 TI - Role of T cell TGF-beta signaling in intestinal cytokine responses and helminthic immune modulation. AB - Colonization with helminthic parasites induces mucosal regulatory cytokines, like IL-10 or TGF-beta, that are important in suppressing colitis. Helminths induce mucosal T cell IL-10 secretion and regulate lamina propria mononuclear cell (LPMC) Th1 cytokine generation in an IL-10-dependent manner in WT mice. Helminths also stimulate mucosal TGF-beta release. As TGF-beta exerts major regulatory effects on T lymphocytes, we investigated the role of T lymphocyte TGF-beta signaling in helminthic modulation of intestinal immunity. T cell TGF-beta signaling is interrupted in TGF-beta receptor II dominant negative (TGF-betaRII DN) mice by T-cell-specific over-expression of a TGF-betaRII DN. We studied LPMC responses in WT and TGF-betaRII DN mice that were uninfected or colonized with the nematode, Heligmosomoides polygyrus. Our results indicate an essential role of T cell TGF-beta signaling in limiting mucosal Th1 and Th2 responses. Furthermore, we demonstrate that helminthic induction of intestinal T cell IL-10 secretion requires intact T cell TGF-beta-signaling pathway. Helminths fail to curtail robust, dysregulated intestinal Th1 cytokine production and chronic colitis in TGF-betaRII DN mice. Thus, T cell TGF-beta signaling is essential for helminthic stimulation of mucosal IL-10 production, helminthic modulation of intestinal IFN-gamma generation and H. polygyrus-mediated suppression of chronic colitis. PMID- 19544492 TI - Chiral analysis of anti-acquired immunodeficiency syndrome drug, 9-(R)-[2 (phosphonomethoxy)propyl]adenine (tenofovir), and related antiviral acyclic nucleoside phosphonates by CE using beta-CD as chiral selector. AB - A new CZE method has been developed for chiral analysis of an important anti acquired immunodeficiency syndrome drug, 9-(R)-[2 (phosphonomethoxy)propyl]adenine ((R)-PMPA, tenofovir), and six related antiviral acyclic nucleoside phosphonates using beta-CD as a chiral selector. The influence of the composition, concentration and pH of the BGE and the type and concentration of chiral selector on enantiomer resolution was investigated. Complete separations of (R,S)-enantiomers of PMPA with very good resolution (R(s)=1.50-3.64) were achieved within a short time (4-15 min) in 20-50 mM sodium borate or sodium tetraborate BGEs, pH 10.0, at 20 mg/mL concentration of beta-CD. (R,S)-enantiomers of five similar PMPA analogs containing purine bases (adenine, diaminopurine or guanine) and hydroxyl or fluor substituents at C3 carbon atom of propyl chain were baseline separated within 10-17 min in 35 mM sodium tetraborate BGE, pH 10.0, at 20 mg/mL beta-CD concentration. Another important antiviral used by acquired immunodeficiency syndrome patients, derived from pyrimidine base cytosine, 1-(S)-[3-hydroxy-2-(phosphonomethoxy)propyl]cytosine (cidofovir), and the (R)-enantiomer of this drug were successfully separated in 50 mM sodium tetraborate BGE, pH 10.5, at 20 mg/mL beta-CD concentration within 45 min. Using the UV-absorption detection at 206 nm, the concentration detection limits of the analyzed acyclic nucleoside phosphonates were determined in the submicromolar to micromolar range (0.15-2.51 microg/mL level). PMID- 19544493 TI - The role of triphenylamine in the stabilization of highly efficient polyfluorene based OLEDs: a model oligomers study. AB - In order to understand the factors responsible for the improved efficiency and stability of organic light-emitting diodes (OLEDs) based on poly(9,9 dioctylfluorene) (PFO) when triphenylamine (TPA) is introduced as lateral fluorene substituent, we synthetize mono-disperse fluorene-thiophene oligomers as model compounds. Their blends with different concentrations of the fluorenone containing oligomer are studied in order to verify if only a reduction of ketonic defect sites or also an impeded energy transfer (ET) towards such sites are responsible for the suppression of the green emission band. We show that the introduction of TPA groups leads specifically both to an antioxidant action and a reduced ET towards residual defect sites, thanks to the environmental micro encapsulation role played by TPA units surrounding the polymer backbone. As a result, the performances and colour stability of a device fabricated with TPA substituted PFO (PFTPA) are strongly improved with respect to standard PFO device prepared in the same conditions. PMID- 19544491 TI - A proteomic workflow for discovery of serum carrier protein-bound biomarker candidates of alcohol abuse using LC-MS/MS. AB - The diagnosis and care of patients with alcohol abuse and dependence is hampered by a lack of sensitive and specific screening and monitoring tests. Proteomics is a good approach to search for biomarkers of alcohol abuse. Serum carrier protein bound proteins have attracted significant interest because they remain a relatively un-mined region of the proteome. In the present study, a proteomic workflow including LC-MS/MS with enrichment of serum carrier protein-bound biomarkers technique was applied to profile the changes in quality and quantity of serum carrier protein-bound proteins for the discovery of novel biomarker candidates of alcohol abuse. In total, 311 proteins identified with high confidence were discovered to be bound to serum carrier proteins. Complement isoforms, Ig fragments, and apolipoprotein family proteins are the main serum carrier-bound proteins. Protein quantification analysis with and without concern as to gender revealed that gender is a critical consideration for biomarker development in alcohol abuse. Identified proteins not previously associated with alcohol abuse include gelsolin, selenoprotein P, serotransferrin, tetranectin, hemopexin, histidine-rich glycoprotein, plasma kallikrein, and vitronectin. Altered abundance of these proteins suggests that they may be potential novel biomarkers for alcohol abuse. PMID- 19544494 TI - IL-23-driven encephalo-tropism and Th17 polarization during CNS-inflammation in vivo. AB - IL-23 but not IL-12 is essential for the development of autoimmune tissue inflammation in mice. Conversely, IL-12 and IL-23 impact on the polarization of Th1 and Th17 cells, respectively. While both polarized T helper populations can mediate autoimmune inflammation, their redundancy in the pathogenesis of EAE indicates that IL-23 exerts its crucial influence on the disease independent of its T helper polarizing capacity. To study the impact of IL-23 and IL-12 on the behavior of encephalitogenic T cells in vivo, we generated BM-chimeric mice in which we can trace individual populations of IL-23 or IL-12 responsive T helper cells during EAE. We observed that T cells, which lack IL-12Rbeta1 (no IL-12 and IL-23 signaling), fail to invade the CNS and do not acquire a Th17 phenotype. In contrast, loss of IL-12 signaling prevents Th1 polarization but does not prevent T-cell entry into the CNS. The loss of IL-12R engagement does not appear to alter T-cell expansion but leads to their accumulation in secondary lymphoid organs. We found that IL-23 licenses T cells to invade the target tissue and to exert their effector function, whereas IL-12 is critical for Th1 differentiation, but does not influence the pathogenic capacity of auto-reactive T helper cells in vivo. PMID- 19544498 TI - Highly regio- and stereoselective synthesis of nine- to twelve-membered cyclic compounds by a Pd0-catalyzed cyclization reaction between allenes with a nucleophilic functionality and organic halides. PMID- 19544497 TI - Pancreato-enteric anastomosis: the duct evagination technique. AB - Pancreatic anastomotic failure remains the most frequent and potentially life threatening complication following Pancreatoduodenectomy. Numerous modifications in the technique of the pancreatoenteric anastomosis have been reported. We suggest a simple modification which involves "evaginating" the cut end of the pancreatic duct. This technique helps avoid a compromise of the pancreatic ductal patency, and by achieving a wide pancreatic ductal opening can facilitate a safer pancreato-enteric anastomosis. In addition, by possibly decreasing the likelihood of post-operative pancreatic ductal stenosis, it has the potential to reduce post Pancreatoduodenectomy pancreatic exocrine insufficiency. The modification acts as an adjunct to an already established technique yielding good results. PMID- 19544499 TI - Efficient reoxidation of palladium by a hybrid catalyst in aerobic palladium catalyzed carbocyclization of enallenes. PMID- 19544501 TI - Asymmetric organocatalytic four-component quadruple domino reaction initiated by oxa-Michael addition of alcohols to acrolein. PMID- 19544500 TI - Preparation of 2-quinolones by sequential Heck reduction-cyclization (HRC) reactions by using a multitask palladium catalyst. AB - One-pot sequential Heck reduction-cyclization (HRC) reactions leading to the synthesis of substituted 2-quinolones have been developed by using a heterogeneous or mixed homogeneous/heterogeneous multitask palladium catalyst with charcoal as a support. The whole sequence occurs under very mild conditions without the need for additives (ligand or base) by taking advantage of the high reactivity of aryldiazonium salts as "super electrophiles". Recycling experiments showed that the reused heterogeneous Pd(0)/C catalyst was not able to promote another HRC sequence but was, however, still highly active for hydrogenation, hydrodehalogenation, as well as hydrogenolysis reactions. PMID- 19544502 TI - Cationic heterocycles as ligands: synthesis and reactivity with anionic nucleophiles of cationic triruthenium clusters containing C-metalated N methylquinoxalinium or N-methylpyrazinium ligands. AB - The cationic cluster complexes [Ru3(CO)10(mu-H)(mu-kappa2N,C-L1Me)]+ (3+; HL1=quinoxaline) and [Ru3(CO)10(mu-H)(mu-kappa2N,C-L2Me)]+ (5+; HL2=pyrazine) have been prepared as triflate salts by treatment of their neutral precursors [Ru3(CO)10(mu-H)(mu-kappa2N,C-Ln)] with methyl triflate. The cationic character of their heterocyclic ligands is responsible for their enhanced tendency to react with anionic nucleophiles relative to that of hydrido triruthenium carbonyl clusters that have neutral N-heterocyclic ligands. These clusters react instantaneously with methyl lithium and potassium tris-sec-butylborohydride (K selectride) to give neutral products that contain novel nonaromatic N heterocyclic ligands. The following are the products that have been isolated: [Ru3(CO)9(mu-H)(mu3-kappa2N,C-L1Me2)] (6; from 3+ and methyl lithium), [Ru3(CO)9(mu-H)(mu3-kappa2N,C-L1HMe)] (7; from 3+ and K-selectride), [Ru3(CO)9(mu H)(mu3-kappa2N,C-L2Me2)] (8; from 5+ and methyl lithium), and [Ru3(CO)9(mu-H)(mu3 kappa2N,C-L2HMe)] (11; from 5+ and K-selectride). Whereas the reactions of 3+ lead to products that arise from the attack of the corresponding nucleophile at the C atom of the only CH group adjacent to the N-methyl group, the reactions of 5+ give mixtures of two products that arise from the attack of the nucleophile at one of the C atoms located on either side of the N-methyl group. The LUMOs and the atomic charges of 3+ and 5+ confirm that the reactions of these clusters with anionic nucleophiles are orbital-controlled rather than charge-controlled processes. The N-heterocyclic ligands of all of these neutral products are attached to the metal atoms in nonconventional face-capping modes. Those of compounds 6-8 have the atoms of a ligand C=N fragment sigma-bonded to two Ru atoms and pi-bonded to the other Ru atom, whereas the ligand of compound 11 has a C-N fragment attached to a Ru atom through the N atom and to the remaining two Ru atoms through the C atom. A variable-temperature 1H NMR spectroscopic study showed that the ligand of compound 7 is involved in a fluxional process at temperatures above -93 degrees C, the mechanism of which has been satisfactorily modeled with the help of DFT calculations and involves the interconversion of the two enantiomers of this cluster through a conformational change of the ligand CH(2) group, which moves from one side of the plane of the heterocyclic ligand to the other, and a 180 degrees rotation of the entire organic ligand over a face of the metal triangle. PMID- 19544503 TI - Synthesis, structure, and bonding of novel homodinuclear cobalt and nickel borylene complexes. AB - Herein we report for the first time full details on the synthesis and structural characterization of novel homodinuclear bridging cobalt and nickel borylene complexes containing bridging carbonyl ligands, an unusual coordination motif rarely before observed for homodinuclear borylene complexes. Furthermore, the homodinuclear nickel complex represents the first instance of a nickel borylene complex. Quantum chemical analyses of charge-density topology, electron localization function (ELF) and natural charges indicate the absence of direct metal-metal bonds in both the cobalt and nickel systems, in contradiction with electron counting. The topology of the Laplacian of the electron density and of the ELF around the bridging boron atom is consistent with a bis-metallo substituted borane situation for the dicobalt system, but with a three-center bonding borylene for the dinickel complex. PMID- 19544504 TI - The AZARYPHOS family of ligands for ambifunctional catalysis: syntheses and use in ruthenium-catalyzed anti-Markovnikov hydration of terminal alkynes. AB - The family of AZARYPHOS (aza-aryl-phosphane) phosphane ligands, containing a phosphine unit and sterically shielded nitrogen lone pairs in the ligand periphery, is introduced as a tool for developing ambifunctional catalysis by the metal center and nitrogen lone pairs in the ligand sphere. General synthetic strategies have been developed to synthesize over 25 examples of structurally diverse (6-aryl-2-pyridyl)phosphanes (ARPYPHOS), (6-alkyl-2-pyridyl)phosphanes (ALPYPHOS), 4,6-disubsituted 1,3-diazin-2-ylphosphanes or 1,3,5-triazin-2 ylphosphanes, quinazolinylphosphanes, quinolinylphosphanes, and others. The scalable syntheses proceed in a few steps. The incorporation of AZARYPHOS ligands (L) into complexes [RuCp(L)(2)(MeCN)][PF(6)] (Cp = cyclopentadienyl) gives catalysts for the anti-Markovnikov hydration of terminal alkynes of the highest known activities. Electronic and steric ligand effects modulate the reaction kinetics over a range of two orders of magnitude. These results highlight the importance of using structurally diverse ligand families in the process of developing cooperative ambifunctional catalysis by a metal and its ligand. PMID- 19544505 TI - Preparation of polyfunctional arylmagnesium, arylzinc, and benzylic zinc reagents by using magnesium in the presence of LiCl. AB - The presence of LiCl considerably facilitates the insertion of magnesium into various aromatic and heterocyclic bromides. Several functional groups, such as OBoc, -OTs, -Cl, -F, -CF(3), -OMe, -NMe(2), and -N(2)NR(2), are well tolerated. The presence of a cyano group leads in some cases to competitive reduction of the organic halide to the corresponding ArH compound. The presence of sensitive groups such as methyl or ethyl ester is tolerated upon in situ trapping of the intermediate magnesium reagent with ZnCl(2). This method can also be applied to the preparation of functionalized benzylic zinc reagents from benzylic chlorides. In the case of di- or tribromoaryl derivatives, directing groups such as -OPiv, OTs, -N(2)NR(2), or -OAc orient the zinc insertion (Zn/LiCl) to the ortho position, while the reaction with Mg/LiCl or Mg/LiCl/ZnCl(2) leads to regioselective insertion into the para-carbon-bromine bond. Large-scale experiments (20-100 mmol) for all of the metalation procedures are described. PMID- 19544506 TI - Imidazolones in diastereoselective cyclization reactions and Cu(II)-catalysed cross-coupling reactions. PMID- 19544507 TI - Molecular nitrides with titanium and Group 13-15 elements. AB - Several heterometallic nitrido complexes were prepared by reaction of the imido nitrido titanium complex [{Ti(eta(5)-C(5)Me(5))(mu-NH)}(3)(mu(3)-N)] (1) with amido derivatives of Group 13-15 elements. Treatment of 1 with bis(trimethylsilyl)amido [M{N(SiMe(3))(2)}(3)] derivatives of aluminum, gallium, or indium in toluene at 150-190 degrees C affords the single-cube amidoaluminum complex [{(Me(3)Si)(2)N}Al{(mu(3)-N)(2)(mu(3)-NH)Ti(3)(eta(5)-C(5)Me(5))(3)(mu(3) N)}] (2) or the corner-shared double-cube compounds [M(mu(3)-N)(3)(mu(3) NH)(3){Ti(3)(eta(5)-C(5)Me(5))(3)(mu(3)-N)}(2)] [M = Ga (3), In (4)]. Complexes 3 and 4 were also obtained by treatment of 1 with the trialkyl derivatives [M(CH(2)SiMe(3))(3)] (M = Ga, In) at high temperatures. The analogous reaction of 1 with [{Ga(NMe(2))(3)}(2)] at 110 degrees C leads to [{Ga(mu(3)-N)(2)(mu(3) NH)Ti(3)(eta(5)-C(5)Me(5))(3)(mu(3)-N)}(2)] (5), in which two [GaTi(3)N(4)] cube type moieties are linked through a gallium-gallium bond. Complex 1 reacts with one equivalent of germanium, tin, or lead bis(trimethylsilyl)amido derivatives [M{N(SiMe(3))(2)}(2)] in toluene at room temperature to give cube-type complexes [M{(mu(3)-N)(2)(mu(3)-NH)Ti(3)(eta(5)-C(5)Me(5))(3)(mu(3)-N)}] [M = Ge (6), Sn (7), Pb (8)]. Monitoring the reaction of 1 with [Sn{N(SiMe(3))(2)}(2)] and [Sn(C(5)H(5))(2)] by NMR spectroscopy allows the identification of intermediates [RSn{(mu(3)-N)(mu(3)-NH)(2)Ti(3)(eta(5)-C(5)Me(5))(3)(mu(3)-N)}] [R = N(SiMe(3))(2) (9), C(5)H(5) (10)] in the formation of 7. Addition of one equivalent of the metalloligand 1 to a solution of lead derivative 8 or the treatment of 1 with a half equivalent of [Pb{N(SiMe(3))(2)}(2)] afford the corner shared double-cube compound [Pb(mu(3)-N)(2)(mu(3)-NH)(4){Ti(3)(eta(5) C(5)Me(5))(3)(mu(3)-N)}(2)] (11). Analogous antimony and bismuth derivatives [M(mu(3)-N)(3)(mu(3)-NH)(3){Ti(3)(eta(5)-C(5)Me(5))(3)(mu(3)-N)}(2)] [M = Sb (12), Bi (13)] were obtained through the reaction of 1 with the tris(dimethylamido) reagents [M(NMe(2))(3)]. Treatment of 1 with [AlCl(2){N(SiMe(3))(2)}(OEt(2))] affords the precipitation of the singular aluminum-titanium square-pyramidal aggregate [{{(Me(3)Si)(2)N}Cl(3)Al(2)}(mu(3) N)(mu(3)-NH)(2){Ti(3)(eta(5)-C(5)Me(5))(3)(mu-Cl)(mu(3)-N)}] (14). The X-ray crystal structures of 5, 11, 13, 14, and [AlCl{N(SiMe(3))(2)}(2)] were determined. PMID- 19544508 TI - Precursor and solvent effects in the nonhydrolytic synthesis of complex oxide nanoparticles for bioimaging applications by the ether elimination (Bradley) reaction. AB - Investigation of the solvent and alkoxide precursor effect on the nonhydrolytic sol-gel synthesis of oxide nanoparticles by means of an ether elimination (Bradley) reaction indicates that the best crystallinity of the resulting oxide particles is achieved on application of aprotic ketone solvents, such as acetophenone, and of smallest possible alkoxide groups. The size of the produced primary particles is always about 5 nm caused by intrinsic mechanisms of their formation. The produced particles, possessing the composition of natural highly insoluble minerals, are biocompatible. Optical characteristics of the perovskite complex oxide nanoparticles can easily be controlled through doping with rare earth cations; for example, by Eu(3+). They can be targeted through surface modification by anchoring the directing biomolecules through a phosphate or phosphonate moiety. Testing of the distribution of Eu-doped BaTiO(3) particles, modified with ethylphosphonic acid, demonstrates their facile uptake by the plants with active fluid transport, resulting finally in their enhanced concentration within the cell membranes. PMID- 19544509 TI - Chemoselective aromatic azido reduction with concomitant aliphatic azide employing Al/Gd triflates/NaI and ESI-MS mechanistic studies. AB - Aluminium and gadolinium triflates catalyze the chemoselective reduction of aromatic azides to the corresponding amines in combination with sodium iodide. This mild chemoselective method has been applied to the synthesis of various aryl amines, C2-azido-substituted pyrrolo[2,1-c][1,4]benzodiazepines, and fused[2,1 b]quinazolinones by an intramolecular azido reduction tandem cyclization reaction. Interestingly, this methodology selectively reduces aryl azides with enhanced yields and proceeds in shorter reaction times than previous strategies. The mechanistic aspects have been investigated and the intermediates associated with this selective transformation have been intercepted and characterized by online monitoring of the reaction by ESI-MS. PMID- 19544510 TI - Role of the support in catalysis: activation of a mononuclear ruthenium complex for ethene dimerization by chemisorption on dealuminated zeolite Y. AB - A set of supported ruthenium complexes with systematically varied ratios of chemisorbed to physisorbed species was formed by contacting cis [Ru(acac)(2)(C(2)H(4))(2)] (I; acac = C(5)H(7)O(2) (-)) with dealuminated zeolite Y. Extended X-ray absorption fine structure (EXAFS) spectra used to characterize the samples confirmed the systematic variation in the loadings of the two supported species and demonstrated that removal of bidentate acac ligands from I accompanied chemisorption to form [Ru(acac)(C(2)H(4))(2)](+) attached through two Ru-O bonds to the Al sites of the zeolite. A high degree of uniformity in the chemisorbed species was demonstrated by sharp bands in the infrared (IR) spectrum characteristic of ruthenium dicarbonyls that formed when CO reacted with the anchored complex. When the ruthenium loading exceeded 1.0 wt % (Ru/Al approximately 1:6), the additional adsorbed species were simply physisorbed. Ethene ligands on the chemisorbed species reacted to form butenes when the temperature was raised to approximately 393 K; acac ligands remained bonded to Ru. In contrast, ethene ligands on the physisorbed complex simply desorbed under the same conditions. The chemisorption activated the ruthenium complex and facilitated dimerization of the ethene, which occurred catalytically. IR and EXAFS spectra of the supported samples indicate that 1) Ru centers in the chemisorbed species are more electron deficient than those in the physisorbed species and 2) Ru-ethene bonds in the chemisorbed species are less symmetric than those in the physisorbed species, which implies the presence of a preferred configuration for the catalytic dimerization. PMID- 19544511 TI - Fabrication of luminescent CdS nanoparticles on short-peptide-based hydrogel nanofibers: tuning of optoelectronic properties. AB - The pH-induced self-assembly of three synthetic tripeptides in water medium is used to immobilize luminescent CdS nanoparticles. These peptides form a nanofibrillar network structure upon gelation in aqueous medium at basic pH values (pH 11.0-13.0), and the fabrication of CdS nanoparticles on the gel nanofiber confers the luminescent property to these gels. Atomic force microscopy, field-emission scanning electron microscopy, and high-resolution transmission electron microscopy clearly reveal the presence of CdS nanoparticles in a well-defined array on the gel nanofibers. This is a convenient way to make organic nanofiber-inorganic nanoparticle hybrid nanocomposite systems. The size of the CdS nanoparticles remains almost same before and after deposition on the gel nanofiber. Photoluminescence (PL) measurement of the CdS nanoparticles upon deposition on the gel nanofibers shows a significant blue shift in the emission spectrum of the nanoparticles, and there is a considerable change in the PL gap energy of the CdS nanoparticles after immobilization on different gel nanofibrils. This finding suggests that the optoelectronic properties of CdS nanoparticles can be tuned upon deposition on gel nanofibers without changing the size of the nanoparticles. PMID- 19544512 TI - Multicomponent supramolecular systems: self-organization in coordination-driven self-assembly. AB - The self-organization of multicomponent supramolecular systems involving a variety of two-dimensional (2 D) polygons and three-dimensional (3 D) cages is presented. Nine self-organizing systems, SS(1)-SS(9), have been studied. Each involves the simultaneous mixing of organoplatinum acceptors and pyridyl donors of varying geometry and their selective self-assembly into three to four specific 2 D (rectangular, triangular, and rhomboid) and/or 3 D (triangular prism and distorted and nondistorted trigonal bipyramidal) supramolecules. The formation of these discrete structures is characterized using NMR spectroscopy and electrospray ionization mass spectrometry (ESI-MS). In all cases, the self organization process is directed by: 1) the geometric information encoded within the molecular subunits and 2) a thermodynamically driven dynamic self-correction process. The result is the selective self-assembly of multiple discrete products from a randomly formed complex. The influence of key experimental variables- temperature and solvent--on the self-correction process and the fidelity of the resulting self-organization systems is also described. PMID- 19544514 TI - The birth of a chemical society. AB - A short description of the events leading to the birth of the Italian Chemical Society is given. The first Italian Chemical Society was established in 1909, and resulted from the merging of the Milan and Rome sections. PMID- 19544513 TI - Synthesis and biophysical studies on 35-deoxy amphotericin B methyl ester. AB - The use of molecular editing in the elucidation of the mechanism of action of amphotericin B is presented. A modular strategy for the synthesis of amphotericin B and its designed analogues is developed, which relies on an efficient gram scale synthesis of various subunits of amphotericin B. A novel method for the coupling of the mycosamine to the aglycone was identified. The implementation of the approach has enabled the preparation of 35-deoxy amphotericin B methyl ester. Investigation of the antifungal activity and efflux-inducing ability of this amphotericin B congener provided new clues to the role of the 35-hydroxy group and is consistent with the involvement of double barrel ion channels in causing electrolyte efflux. PMID- 19544515 TI - Electrochemical genotyping by using two ferrocene/isoquinoline-connected DNA probes with different redox potentials on a single electrode. PMID- 19544516 TI - Design of tRNA(Lys)3 ligands: fragment evolution and linker selection guided by NMR spectroscopy. AB - A fragment-based approach for the synthesis of ligands of tRNA(Lys) (3), the HIV reverse-transcription primer, is described. The use of NMR spectroscopy has proved to be very useful in this approach, not only to detect low-affinity complexes between small compounds and RNA, but also to provide information on their binding mode and on the way they can be connected. This NMR-spectroscopy guided analysis enabled us to design micromolar ligands after the optimisation and connection of millimolar fragments with an appropriate linker. The influence of the linker region on the binding affinity and selectivity outlines the importance of having a flexible assemblage strategy with a variety of linkers in such an approach. PMID- 19544517 TI - Emissive or nonemissive? A theoretical analysis of the phosphorescence efficiencies of cyclometalated platinum(II) complexes. AB - We herein report a theoretical analysis based on a density functional theory/time dependent density functional theory (DFT/TDDFT) approach to understand the different phosphorescence efficiencies of a family of cyclometalated platinum(II) complexes: [Pt(NCN)Cl] (1; NCN = 1,3-bis(2-pyridyl)phenyl(-)), [Pt(CNN)Cl] (2; CNN = 6-phenyl-2,2'-bipyridyl(-)), [Pt(CNC)(CNPh)] (3; CNC = 2,6 diphenylpyridyl(2-)), [Pt(R-CNN)Cl] (4; R-CNN = 3-(6'-(2''-naphthyl)-2' pyridyl)isoquinolinyl(-)), and [Pt(R-CNC)(CNPh)] (5; R-CNC = 2,6-bis(2' naphthyl)pyridyl(2-)). By considering both the spin-orbit coupling (SOC) and the electronic structures of these complexes at their respective optimized singlet ground (S(0)) and first triplet (T(opt)(1)) excited states, we were able to rationalize the experimental findings that 1) 1 is a strong emitter while its isomer 2 is only weakly emissive in CH(2)Cl(2) solution at room temperature; 2) although the cyclometalated ligand of 3 has a higher ligand-field strength than that of 1, 3 is nonemissive in CH(2)Cl(2) solution at 298 K; and 3) extension of pi conjugation at the lateral aryl rings of the cyclometalated ligands of 2 and 3 to give 4 and 5, respectively, leads to increased emission quantum yields under the same conditions. We found that Jahn-Teller and pseudo-Jahn-Teller effects are operative in complexes 2 and 3, respectively, on going from the optimized S(0) ground state to the optimized T(opt)(1) excited state, and thus lead to large excited-state structural distortions and hence fast nonradiative decay. Furthermore, a strong-field ligand may push the two different occupied d orbitals so far apart that the SOC effect is small and the radiative decay rate is slow. This work is an example of electronic-structure-driven tuning of the phosphorescence efficiency, and the DFT/TDDFT approach is demonstrated to be a versatile tool for the design of phosphorescent materials with target characteristics. PMID- 19544518 TI - Biological profiling of anti-HIV agents and insight into CCR5 antagonist binding using in silico techniques. AB - Molecular requirements and determinants for efficient binding to CCR5 were interpreted by computational techniques based on comparative receptor structure modeling, advanced 3D-QSAR, docking, and shape-based virtual screening of commercially available entry blockers. Results of this study may be valuable for predicting new HIV entry-blocking leads.Acquired immune deficiency syndrome (AIDS) is responsible for more than 31 million deaths, and many more people are affected by this disease worldwide. Novel ligands that are capable of blocking virus-cell fusion are emerging as promising candidate molecules against HIV-1 infection because they have the promise to overcome the major drawbacks of classical highly active antiretroviral (HAART) drugs. However, structure-based design continues to be hampered owing to the paucity of experimentally determined 3D information about HIV-1 cell-surface co-receptors. Using computational techniques based on comparative receptor structure modeling, advanced 3D-QSAR, and protein-ligand docking, we present recent results that define updated molecular requirements and determinants for efficient binding of small-molecule ligands to CCR5, a principal biological target for HIV entry blockers. These results are compared with shape- and property-based virtual screening results for commercially available entry blockers, and will be valuable for predicting new HIV entry-blocking leads. PMID- 19544519 TI - Nonmetal-mediated fragmentation of P4: isolation of P1 and P2 bis(carbene) adducts. PMID- 19544520 TI - Atropisomerism of alpha,beta-unsaturated amidines: stereoselective synthesis by catalytic cascade reaction and optical resolution. PMID- 19544521 TI - Insights into the geometrical features underlying beta-O-GlcNAc glycosylation: water pockets drastically modulate the interactions between the carbohydrate and the peptide backbone. PMID- 19544522 TI - Metal-catalyzed carboxylation of organometallic reagents with carbon dioxide. PMID- 19544523 TI - A "dual-function" photocage releasing nitric oxide and an anthrylmethyl cation with a single wavelength light. PMID- 19544525 TI - An examination of binding motifs associated with inter-particle interactions between facetted nano-crystals of acetylsalicylic acid and ascorbic acid through the application of molecular grid-based search methods. AB - Grid-based intermolecular search methods using atom-atom force fields are used to assess the structural nature of potential crystal-crystal interfacial binding associated with the examination of representative pharmaceutical formulation components, viz acetylsalicylic acid (aspirin) and ascorbic acid (vitamin C). Molecular models of nano-sized molecular clusters for these two compounds, shaped in accordance with an attachment energy model of the respective particle morphologies, are constructed and used together with a grid-based search method to model the likely inter-particle interactions. The most-stable, mutual alignments of the respective nano-clusters based on their interaction energies are identified in the expectation that these are indicative of the most likely inter-particle binding configurations. The stable inter-particle binding configurations identified reveal that the number of interfacial hydrogen bonds formed between the binding particles is, potentially, an important factor in terms of the stability of inter-particle cohesion. All preferred inter-particle alignments are found to involve either the (1 0 0) or the (1 1 0) face of aspirin crystals interacting with a number of the growth forms of ascorbic acid. Four main types of interfacial hydrogen bonds are found to be associated with inter particle binding and involve acceptor-donor interactions between hydroxyl, carbonyl, ester and lactone acceptor groups and hydroxyl donor groups. This hydrogen bonding network is found to be consistent with the surface chemistry of the interacting habit faces with, in general, the number of hydrogen bonds increasing for the more stable alignments. The likely usefulness of this approach for predicting solid-state formulation properties is reviewed. PMID- 19544524 TI - Effect of fiber distribution and realignment on the nonlinear and inhomogeneous mechanical properties of human supraspinatus tendon under longitudinal tensile loading. AB - Tendon exhibits nonlinear stress-strain behavior that may be partly due to movement of collagen fibers through the extracellular matrix. While a few techniques have been developed to evaluate the fiber architecture of other soft tissues, the organizational behavior of tendon under load has not been determined. The supraspinatus tendon (SST) of the rotator cuff is of particular interest for investigation due to its complex mechanical environment and corresponding inhomogeneity. In addition, SST injury occurs frequently with limited success in treatment strategies, illustrating the need for a better understanding of SST properties. Therefore, the objective of this study was to quantitatively evaluate the inhomogeneous tensile mechanical properties, fiber organization, and fiber realignment under load of human SST utilizing a novel polarized light technique. Fiber distributions were found to become more aligned under load, particularly during the low stiffness toe-region, suggesting that fiber realignment may be partly responsible for observed nonlinear behavior. Fiber alignment was found to correlate significantly with mechanical parameters, providing evidence for strong structure-function relationships in tendon. Human SST exhibits complex, inhomogeneous mechanical properties and fiber distributions, perhaps due to its complex loading environment. Surprisingly, histological grade of degeneration did not correlate with mechanical properties. PMID- 19544526 TI - Biological indicators of prognosis in Ewing's sarcoma: an emerging role for lectin galactoside-binding soluble 3 binding protein (LGALS3BP). AB - Starting from an experimental model that accounts for the 2 most important adverse processes to successful therapy of Ewing's sarcoma (EWS), chemoresistance and the presence of metastasis at the time of diagnosis, we defined a molecular signature of potential prognostic value. Functional annotation of differentially regulated genes revealed 3 major networks related to cell cycle, cell-to-cell interactions and cellular development. The prognostic impact of 8 genes, representative of these 3 networks, was validated in 56 EWS patients. High mRNA expression levels of HINT1, IFITM2, LGALS3BP, STOML2 and c-MYC were associated with reduced risk to death and lower risk to develop metastasis. At multivariate analysis, LGALS3BP, a matricellular protein with a role in tumor progression and metastasis, was the most important predictor of event-free survival and overall survival. The association between LGALS3BP and prognosis was confirmed at protein level, when expression of the molecule was determined in tumor tissues but not in serum, indicating a role for the protein at local tumor microenvironment. Engineered enhancement of LGALS3BP expression in EWS cells resulted in inhibition of anchorage independent cell growth and reduction of cell migration and metastasis. Silencing of LGALS3BP expression reverted cell behavior with respect to in vitro parameters, thus providing further functional validation of genetic data obtained in clinical samples. Thus, we propose LGALS3BP as a novel reliable indicator of prognosis, and we offer genetic signatures to the scientific communities for cross-validation and meta-analysis, which are indispensable tools for a rare tumor such as EWS. PMID- 19544527 TI - The human 1-8D gene (IFITM2) is a novel p53 independent pro-apoptotic gene. AB - The human 1-8 interferon inducible gene family consists of at least 3 functional genes; 9-27, 1-8D and 1-8U, which are all linked on an 18-kb fragment of chromosome 11 and are highly homologous. It has recently been shown by us and others that the 1-8D gene is overexpressed in colon carcinoma. Here, we show, by sequence comparison of the 1-8D in pairs of tumor/normal colon tissues, the existence of 6 different alleles, containing single-nucleotide polymorphisms with no mutations. Transformation assays revealed a possible role for the 1-8D gene as a transformation inhibitor. Further, transient expression of the human 1-8D gene in multiple mammalian cell lines showed accumulation of cells in the G1 phase followed by elevation in the subG1 phase. SubG1 elevation was confirmed as apoptosis by Annexin-V binding assays and transferase-mediated dUTP nick end labeling assays. Moreover, knock-down of 1-8D provided partial protection from Etoposide and UV-induced apoptosis. The induction of apoptosis by 1-8D is dependent on caspase activities but not on p53 expression. Although 1-8D induces apoptosis independently of p53, p53 expression downregulates 1-8D protein expression. Our data suggest a role for the 1-8D gene as a novel pro-apoptotic gene that will provide new insights into the regulated cellular pathways to death. PMID- 19544529 TI - Overexpression of low-density lipoprotein receptor and lipid accumulation in intestinal polyps in Min mice. AB - Apc-deficient Min mice feature low expression of lipoprotein lipase (LPL), high concentration of serum triglyceride (TG), fatty change of the liver and large numbers of intestinal polyps. We have reported that induction of LPL expression reduces serum lipid, especially TG, improves fatty change of the liver and inhibits intestinal polyp formation in the mice. In this study, fatty change/lipid accumulation in intestinal mucosa and polyps in Min mice were analyzed by Oil-red O staining and electron microscopy. A number of large lipid droplets were found in the epithelia of the upper part of polyps. On the other hand, small lipid droplets were only slightly observed at the tip of the villi in non-tumoros parts of the small intestine of Min mice and in the villi of wild type mice. Moreover, low-density lipoprotein receptor (LDLR) was overexpressed in the area where lipid droplets were observed. The expression levels of LDLR mRNA in the intestinal polyps of Min mice were approximately 3 times higher compared to those in the non-tumoros parts. Remarkable expression of cyclooxygenase-2 was mainly distributed in stromal cells and some in epithelial cells. It is speculated that lipid accumulation in the intestinal polyps may play an important role in intestinal polyp formation in Apc-deficient mice. PMID- 19544530 TI - FoxP3+ regulatory T cells are distinct from leukemia cells in HTLV-1-associated adult T-cell leukemia. AB - Human T-lymphotropic virus type 1 (HTLV-1) is the causative agent of adult T-cell leukemia/lymphoma (ATLL). It has been postulated that ATLL cells might act as regulatory T cells (T(regs)) which, in common with ATLL cells, express both CD25 and FoxP3, and so contribute to the severe immune suppression typical of ATLL. We report here that the frequency of CD25(+) cells varied independently of the frequency of FoxP3(+) cells in both a cross-sectional study and in a longitudinal study of 2 patients with chronic ATLL. Furthermore, the capacity of ATLL cells to suppress proliferation of heterologous CD4(+)CD25(-) cells correlated with the frequency of CD4(+) FoxP3(+) cells but was independent of CD25 expression. Finally, the frequency of CD4(+)FoxP3(+) cells was inversely correlated with the lytic activity of HTLV-1-specific CTLs in patients with ATLL. We conclude that ATLL is not a tumor of FoxP3(+) regulatory T cells, and that a population of FoxP3(+) cells distinct from ATLL cells has regulatory functions and may impair the cell-mediated immune response to HTLV-1 in patients with ATLL. PMID- 19544531 TI - Cotransplantation of placental mesenchymal stromal cells enhances single and double cord blood engraftment in nonobese diabetic/severe combined immune deficient mice. AB - Limited cell numbers in a unit restricts cord blood transplantation (CBT) in adults. We evaluated whether cotransplantation of placental mesenchymal stromal cells (MSCs) would enhance engraftment. Plastic adherent cells from placenta demonstrated typical characteristics of MSCs. In six individual experiments, 4 cohorts of 24 nonobese diabetic/severe combined immune deficient (NOD/SCID) mice were evaluated. Cohort 1 received 5 x 10(4) CD34+ cells from unit (U) one (SCBT); cohort 2 received 5 x 10(4) CD34+ cells from U1 + 4 x 10(4) MSCs (SCBT+MSCs); cohort 3 received 2.5 x 10(4) CD34+ cells from U1 + 2.5 x 10(4) CD34+ cells from U2 (double cord blood transplant [DCBT]); cohort 4 received 2.5 x 10(4) CD34+ cells from U1 + 2.5 x 10(4) CD34+ cells from U2 + 4 x 10(4) MSCs (DCBT+MSCs). Hematopoietic engraftment evaluated after 6 to 8 weeks, was similar in recipients of SCBT and DCBT. MSC cotransplantation demonstrated enhanced engraftment in DCBT (51.8 +/- 6.8% versus 14.9 +/- 6.5%; p = .04) with an increased trend in SCBT (48.7 +/- 7.7% versus 17.5 +/- 6.1%; p = .07). In DCBT, cotransplantation of placental MSCs reduced single cord dominance. Self-renewal capacity was assessed by serial transplantation in secondary recipients infused with engrafted human cells from primary mice transplanted with or without MSCs. In secondary transplant experiments, 13 of 17 evaluable mice engrafted at levels of 1% to 6.5%. Despite enhanced engraftment in primary mice, long-term engraftment capacity was unaltered with MSC cotransplantation. Imaging studies showed MSCs migrated to pelvic region and improved cord blood (CB) CD34+ homing. Cotransplantation of placental MSCs enhanced cord blood engraftment and may act by improving homing of CD34+ cells. PMID- 19544528 TI - Serum cholesterol levels in relation to the incidence of cancer: the JPHC study cohorts. AB - An inverse association between serum total cholesterol and cancer mortality cast a controversy for cause or result of low cholesterol on cancer risk. Therefore, we examined a total of 33,368 Japanese men and women aged 40-69 years, who were free of prior diagnosis of cancer and cardiovascular disease, undertook serum total cholesterol measurement and completed a food frequency questionnaire between 1990 and 1994. They were followed to ascertain incident total and major sites of cancer until the end of 2004 to examine sex-specific associations between cholesterol and cancer risk by incident time, stage and virus infection. After 412,714 person-years of follow-up, 2,728 incident cancers were documented. Serum total cholesterol levels were inversely associated with risk of total cancer in men, with strong inverse associations with stomach cancer in men and liver cancer in both sexes. After exclusion for first 3-year incident cases and advanced cases with metastasis, the inverse association diminished for total and stomach cancers but remained for liver cancer. The multivariable hazard ratios (95% CI) for serum total cholesterol <4.14 mmol/l versus 4.65-5.16 mmol/l were 1.15(0.92-1.43); p-trend across the overall cholesterol categories = 0.25 for total cancer and 1.18(0.79-1.75), p-trend = 0.04 for stomach cancer and 5.12(1.65 15.9), p-trend = 0.0011 for liver cancer in men, and 5.73(1.57-20.9), p-trend = 0.0007 for liver cancer in women. The sustained excess risk of liver cancer associated with low cholesterol was observed regardless of hepatitis-C-virus infection and drinking habits. Although the inverse association for liver cancer remained to be examined further, our findings do not support that low serum total cholesterol levels increase risks of total cancer and other major sites. PMID- 19544532 TI - Birth of parthenote mice directly from parthenogenetic embryonic stem cells. AB - Mammalian parthenogenetic embryos are not viable and die because of defects in placental development and genomic imprinting. Parthenogenetic ESCs (pESCs) derived from parthenogenetic embryos might advance regenerative medicine by avoiding immuno-rejection. However, previous reports suggest that pESCs may fail to differentiate and contribute to some organs in chimeras, including muscle and pancreas, and it remains unclear whether pESCs themselves can form all tissue types in the body. We found that derivation of pESCs is more efficient than of ESCs derived from fertilized embryos, in association with reduced mitogen activated protein kinase signaling in parthenogenetic embryos and their inner cell mass outgrowth. Furthermore, in vitro culture modifies the expression of imprinted genes in pESCs, and these cells, being functionally indistinguishable from fertilized embryo-derived ESCs, can contribute to all organs in chimeras. Even more surprisingly, our study shows that live parthenote pups were produced from pESCs through tetraploid embryo complementation, which contributes to placenta development. This is the first demonstration that pESCs are capable of full-term development and can differentiate into all cell types and functional organs in the body. PMID- 19544533 TI - Structural study of the salivary glands of Anocentor nitens (Acari: Ixodidae) during the feeding cycle. AB - The salivary glands of Anocentor nitens (Neumann,1897) occur in pairs and are located in the anterolateral region of the general cavity, with milky white color and approximately equal sizes. They consist of a secretory portion and an excretion duct. In some glandular acini, all the cells had a basophilic appearance they were stained by hematoxylin, whereas others presented cells with different staining affinities. In this work, we describe the variations observed in these glands during the feeding cycle of ticks [after feeding (0 h) and successively at 24, 48, 72, 96, 120, and 144 h]. The cells stained by hematoxylin were shown to be more reactive to Alcian blue, thus demonstrating the presence of acid glycosaminoglycans, whereas those stained using eosin presented weak or no reaction. A strong reaction was found by the use of the periodic acid-Schiff (PAS) technique, thereby suggesting the presence of glycogen and/or glycoconjugates containing hexose, confirmed by using salivary amylase before PAS, with partial destaining of the slides. Continuing presence of residual staining in these cells suggests the presence of glycoconjugates containing hexose. Cells with nuclei of circular outline and few granules (of different sizes) were found in type II acini, 72 h after collection. Type I acini presented wide lumina and walls composed of larger numbers of cells of cubic to cylindrical shape. The pronounced degranulation shown in this study over the course of the feeding cycle was associated with the release of substances for oviposition. PMID- 19544534 TI - Self-assembled monolayers of DNA on cysteamine modified Au(111) surface: Atomic force microscopy study. AB - The lambda-DNA molecules self-assemble on cysteamine-modified gold (111) surface to form flat-lying self-assembled monolayers (SAMs). The formation kinetics of such DNA SAMs is studied by atomic force microscopy (AFM). AFM results show that DNA molecules do not arrange themselves on cysteamine-modified gold (111) surface into a well-ordered monolayer. It is also found that the surface density of DNA monolayer does not increase as the DNA concentration increases. The high temperature of DNA solution and the immersing in ultrapure water produce some obvious DNA bundles. Whereas divalent cations in DNA solution result in the formation of more compact DNA films. The obtained information may be useful for practical application of the DNA films and further theoretical studies. PMID- 19544535 TI - Predictive modeling in prostate cancer. Conclusions and reflections. AB - Validated prognostic factors are an integral part of any cancer diagnosis. The available proliferation of new markers and randomized clinical trial results leads to a complex decision making looking for optimal treatment and outcome results. Advanced mathematic models have been developed to weigh each specific prognostic factor into a single outcome number in nomograms modeled for specific predictive accuracy in key phases of the treated history of the disease. Despite this progress, it is important to realize that the results are an outcome stratification for groups rather than for the individual patient. It is clear that in the tumor prognosis, host and environmental factors need to be evaluated before any clinical decision is made. Cancer 2009;115(13 suppl):3160-2. (c) 2009 American Cancer Society. PMID- 19544536 TI - Zero in on high-priority breast cancer trials. PMID- 19544537 TI - The diagnostic and prognostic utility of positron emission tomography/computed tomography-based follow-up after radiotherapy for head and neck cancer. AB - BACKGROUND: The detection of subclinical head and neck cancer recurrence or a second primary tumor may improve survival. In the current study, the authors investigated the clinical value of a follow-up program incorporating serial (18)F fluorodeoxyglucose-positron emission tomography integrated with computed tomography (PET/CT) in the detection of recurrent disease in patients with head and neck cancer. METHODS: A total of 240 PET/CT scans were reviewed in 80 patients with head and neck cancer who were treated with radiotherapy (RT) from July, 2005 through August, 2007. All patients were followed with clinical examination, PET/CT, and correlative imaging for a minimum of 11 months (median follow-up, 21 months). RESULTS: The sensitivity, specificity, and positive and negative predictive values of PET/CT-based follow-up for detecting locoregional recurrence were 92%, 82%, 42%, and 98%, respectively. Corresponding values for distant metastases or second primary tumors were 93%, 96%, 81%, and 98%, respectively. Eight patients (10%) developed disease recurrences or second primary tumors that were amenable to salvage surgery with negative surgical margins. The 2-year progression-free survival and 2-year overall survival rates were significantly different between patients who had a negative and those with a positive PET/CT result within 6 months of the completion of RT (93% vs 30% [P<.001] and 100% vs 32% [P<.001], respectively). CONCLUSIONS: Although post therapy follow-up using PET/CT is reportedly associated with a high false positive rate in the irradiated head and neck, PET/CT appears to be a highly sensitive technique for the detection of recurrent disease. Furthermore, negative PET/CT results within 6 months of the completion of RT offer significant prognostic value. PMID- 19544538 TI - Nomograms for staging, prognosis, and predicting treatment outcomes. AB - Prostate cancer is a heterogeneous disease with a wide prognostic spectrum and a variety of treatment options. Such a complex clinical scenario has led to uncertainty in risk assessment and prediction of outcome. Nomograms have served as scientific formulas designed to maximize the predictive accuracy. The use of nomograms in prostate cancer has been applied to many clinical states and outcomes and has provided the most accurate predictions. Cancer 2009;115(13 suppl):3107-11. (c) 2009 American Cancer Society. PMID- 19544540 TI - The current state of brachytherapy nomograms for patients with clinically localized prostate cancer. AB - For men diagnosed with clinically localized prostate cancer, definitive therapy with radical prostatectomy, external beam radiation therapy, or brachytherapy offers a high chance of cure. Currently, there are insufficient data to recommend 1 treatment approach over another, leaving physicians and patients to decide based on their own biases and preferences. Prediction tools, such as nomograms and probability tables, have been created as decision aids to facilitate patient counseling and decision making. Nomograms in particular can assess the therapeutic efficacy of a given therapy by providing individualized estimates of the risk of failure after treatment. The authors performed a comprehensive literature review to identify nomograms assessing the efficacy of brachytherapy in patients with clinically localized prostate cancer, and found a paucity of such models. Analysis of currently available brachytherapy nomograms reveals suboptimal predictive power compared with models based on other treatment modalities. The purpose of this review is to spur development of new and more accurate prediction tools for predicting outcomes after brachytherapy, offering physicians and patients the opportunity to equally assess the efficacy of all available treatment modalities for clinically localized prostate cancer. Cancer 2009;115(13 suppl):3121-7. (c) 2009 American Cancer Society. PMID- 19544539 TI - Predictive models in external beam radiotherapy for clinically localized prostate cancer. AB - Predictive models are being used increasingly in effort to allow physician and patient expectations to be aligned with outcomes that are based on available data. Most predictive models for men who receive external beam radiotherapy for clinically localized prostate cancer are based on Gleason score, clinical tumor classification, and prostate-specific antigen (PSA) levels. More sophisticated models also have been developed that incorporate treatment-related variables, such as the dose of radiation and the use of androgen-deprivation therapy. Most of the predictive models applied to prostate cancer were derived using PSA recurrence rates as the major endpoint, but clinical endpoints have been incorporated increasingly into predictive models. Biomarkers also are increasingly being added to predictive models in an effort to strengthen them. The Radiation Therapy Oncology Group (RTOG) has completed studies on a wide range of markers using tissue from 2 phase 3 trials (RTOG 8610 and 9202). To date, preliminary assessments of p53; DNA ploidy; p16/retinoblastoma 1 protein; Ki-67; mouse double-minute p53 binding protein homolog; Bcl-2/Bcl-2-associated X protein; cytosine, adenine, and guanine repeats; cyclooxygenase-2; signal transducer and activator of transcription 3; cytochrome P450 3A4; and protein kinase A have been completed. Although they are not ready for widespread, routine use, there are reasons to believe that future models will combine these markers with traditional pretreatment and treatment-related variables and will improve our ability to predict outcome and select the optimal treatment. Cancer 2009;115(13 suppl):3112-20. (c) 2009 American Cancer Society. PMID- 19544541 TI - Predictive models in palliative care. AB - It is important to identify prognostic and predictive factors concerning both life expectancy and quality of life in palliative care patients to facilitate ethical, clinical, and organizational decisions, but also to use resources in the best possible way. The authors reviewed the literature to identify the major factors that can predict survival of patients with solid tumors. They found only a few prospective assessments of prognostic factors. Clinical prognostic/predictors of survival based on physician's and/or nurse's judgment, performance status, dyspnea at rest, anorexia, dysphagia, or delirium are all considered to be of primary importance. Despite several contrasting findings, it is generally agreed that the type and site of the primary tumor and metastasis, psychosocial factors, and quality of life should be considered secondary to the organic effects in the final stages of life. Leukocytosis, lymphocytopenia, and elevated C-reactive protein are all reported to have prognostic significance, and low serum albumin and high lactate dehydrogenase levels must also be taken into consideration. Cancer 2009;115(13 suppl):3128-34. (c) 2009 American Cancer Society. PMID- 19544542 TI - Predictive models of toxicity in external radiotherapy: dosimetric issues. AB - Dose-volume modeling of late and acute toxicity in radiotherapy for prostate cancer is a rapidly evolving field of investigation. The availability of individual, 3-dimensional dose distribution and dose-volume histograms (DVHs) permits the quantitative assessment of dose-volume relations for specific endpoints by investigating the correlation between individual dose-volume data and clinical outcomes. These studies often entail a huge effort in collecting data from large populations that have been followed properly for long time. The rectum is the most investigated organ, especially concerning late bleeding, and a good consensus regarding serial-like behavior for this endpoint comes from various investigations. Concerning the bladder, the existence of a clear dose effect when the organ is wholly or partially irradiated is well known. Concerning erectile dysfunctions, the wide use of hormone therapy and drugs against impotence suggests that large, prospectively scored populations will be necessary to definitively assess dose-volume relations. Bowel irradiation during prostate cancer radiotherapy occurs during pelvic lymph node irradiation, and severe acute toxicity has yet to be modeled clearly. Evidence of a correlation between the DVH of the intestinal cavity and bowel toxicity recently was reported, providing important information about optimal dose-volume constraints to be used during whole-pelvis irradiation with intensity-modulated radiotherapy. Cancer 2009;115(13 suppl):3135-40. (c) 2009 American Cancer Society. PMID- 19544543 TI - Predictive models of toxicity with external radiotherapy for prostate cancer: clinical issues. AB - The objective of the current study was to analyze the state of the art and present limitations of available predictive clinical models (when available) estimating the risk of genitourinary tract and small bowel complications, erectile dysfunction, and acute and late symptoms of the rectal syndrome caused by prostate cancer external irradiation. An analysis of the literature indicated that very limited attention has been devoted to the development of "integrated," patient-tailored, user-friendly, and clinically usable tools for the prediction of external beam radiotoxicity. In this article, the authors reported on the multivariate correlation between late genitourinary and gastrointestinal toxicities and clinical/dosimetric risk factors, as well as on the first set of nomograms developed to predict acute and late rectal side effects. At the present state of knowledge, the use of nomograms as predictive instruments of radiotoxicity appears to be particularly attractive for several main reasons. They are "user friendly" and easily developed using the results of multivariate analyses, as they weigh the combined effects of multiple independent factors found to be correlated with the selected clinical endpoint. The integrated evaluation of clinical and dosimetric parameters in the single patient can help to provide a tailored probability of the specific outcome considered. Predicting a high probability of toxicity could avoid unnecessary daily costs for the individual patient in terms of quality of life modification during and after treatment, helping patients in the decision-making process of choosing the best individual, quality of life-related treatment, and clinicians in better tailoring the treatment to patient's characteristics. Cancer 2009;115(13 suppl):3141-9. (c) 2009 American Cancer Society. PMID- 19544545 TI - Development of a prostate cancer metagram: a solution to the dilemma of which prediction tool to use in patient counseling. AB - Many treatment options are available to the human with clinically localized prostate cancer, including surgery, radiation, and even active surveillance. To the authors' knowledge, there is no consensus on the optimal management of this patient population, with most clinicians tending to recommend the treatment with which they are most familiar. Effective patient counseling allowing informed decision making can be best achieved with a formalized system that offers accurate predictions of outcomes for all available treatment approaches. The authors organized the currently available prostate cancer prediction tools toward the formation of a metagram that can be used to tailor management to the individual patient. A comprehensive review of the literature was performed to identify published prediction tools intended for use in prostate cancer. Tools were categorized by a combination of treatment modality and the outcome being predicted, and incorporated into a metagram constructed of 16 different treatment options and 10 outcomes related to cancer control, survival, and morbidity. A search of the literature revealed 44 prostate cancer prediction tools that assessed at least 1 of the 160 treatment/outcome combinations that comprise the metagram. Only 31 cells of the metagram were populated with currently available tools. Prediction tools offer the most accurate estimates of outcomes in prostate cancer, but their current role in patient counseling is complicated by the large number of existing tools, as well as a lack of comparative data. To address this, the authors incorporated the most relevant prediction tools currently available into a prostate cancer metagram that may offer evidence-based and individualized predictions for multiple endpoints after all available treatment options in clinically localized prostate cancer. The metagram also reveals areas of deficiency in the current catalog of prediction tools. Many more prediction tools are needed. Cancer 2009;115(13 suppl):3039-45. (c) 2009 American Cancer Society. PMID- 19544544 TI - Prediction of sexual function after radical prostatectomy. AB - Radical prostatectomy (RP) is a commonly used procedure in the treatment of clinically localized prostate cancer. For this report, the authors critically analyzed the factors associated with recovery of erectile function after surgery. A systematic review of the literature using the Medline and CancerLit databases was conducted. Keywords for the literature search included prostate cancer, radical prostatectomy, erectile dysfunction, impotence, treatment, and prophylaxis. Accurate patient selection (based on patient age, preoperative erectile function, and comorbidity profile) and adequate surgical technique (ie, the preservation of neurovascular bundles) were the major determinants of postoperative erectile function. Moreover, better results were achieved when an appropriate pharmacologic treatment using either oral or local approaches was given. Therefore, the authors concluded that, if patients are stratified correctly according to preoperative, intraoperative, and postoperative factors, then a satisfactory functional recovery may be expected after surgery. For these reasons, an ideal multivariate model predicting the restoration of erectile function after surgery should include patient, surgeon, and postsurgical treatment variables. The authors also concluded that the stratification of patients with regard to their risk of developing erectile dysfunction after surgery was feasible based on several parameters, which should be taken into account for correct patient treatment and counseling. To address this objective, accurate tools for predicting the likelihood of complete functional recovery after surgery are needed. Cancer 2009;115(13 suppl):3150-9. (c) 2009 American Cancer Society. PMID- 19544546 TI - Genomic approaches to outcome prediction in prostate cancer. AB - Prostate cancer remains a common cause of cancer death in men. Applications of emerging genomic technologies to high-quality prostate cancer models and patient samples in multiple contexts have made significant contributions to our molecular understanding of the development and progression of prostate cancer. Genomic analysis of DNA, RNA, and protein alterations allows for the global assessment of this disease and provides the molecular framework to improve risk classification, outcome prediction, and development of targeted therapies. In this review, the author focused on highlighting recent work in genomics and its role in evaluating molecular modifiers of prostate cancer risk and behavior and the development of predictive models that anticipate the risk of developing prostate cancer, prostate cancer progression, and the response of prostate cancer to therapy. This framework has the exciting potential to be predictive and to provide personalized and individual treatment to the large number of men diagnosed with prostate cancer each year. Cancer 2009;115(13 suppl):3046-57. (c) 2009 American Cancer Society. PMID- 19544548 TI - Decision support systems for morphology-based diagnosis and prognosis of prostate neoplasms: a methodological approach. AB - Recent advances in computer and information technologies have allowed the integration of both numeric and non-numeric data, that is, descriptive, linguistic terms. This has led at 1 end of the spectrum of technology development to machine vision based on image understanding and, at the other, to decision support systems. This has had a significant impact on our capability to derive diagnostic and prognostic information from histopathological material with prostate neoplasms. Cancer 2009;115(13 suppl):3068-77. (c) 2009 American Cancer Society. PMID- 19544547 TI - Biomolecular markers of outcome prediction in prostate cancer. AB - Prostate cancer has a variable clinical outcome and, therefore, there is a clear need for novel molecular markers that are specifically associated with biologically aggressive disease to improve staging and prognostication and also to provide mechanistic information to facilitate treatment selection. Different candidate biomarkers have been identified that are linked to patient prognosis and/or response to specific treatments. Such molecules are involved in diverse cellular processes (including cell cycle regulation, cell death and apoptosis, signal transduction, cell adhesion, and angiogenesis) within which aberrant activity of several regulatory pathways has been seen in prostate cancer. Although the number of molecular markers continues to grow, mainly because of the advent of high-throughput methods, more work needs to be done to develop uniform standards for their characterization to enable comparison of markers across studies. Moreover, a rate-limiting step in the development of molecular markers is large-scale clinical assessment and their evaluation in the context of prediction model improvement. In fact, thus far, only a few studies have tested and demonstrated whether the addition of new biological markers improves the accuracy of standard clinical models (nomograms) in predicting biochemical progression in patients with clinically localized prostate cancer who underwent radical prostatectomy. Cancer 2009;115(13 suppl):3058-67. (c) 2009 American Cancer Society. PMID- 19544550 TI - Techniques and predictive models to improve prostate cancer detection. AB - The use of prostate-specific antigen (PSA) as a screening test remains controversial. There have been several attempts to refine PSA measurements to improve its predictive value. These modifications, including PSA density, PSA kinetics, and the measurement of PSA isoforms, have met with limited success. Therefore, complex statistical and computational models have been created to assess an individual's risk of prostate cancer more accurately. In this review, the authors examined the methods used to modify PSA as well as various predictive models used in prostate cancer detection. They described the mathematical underpinnings of these techniques along with their intrinsic strengths and weaknesses, and they assessed the accuracy of these methods, which have been shown to be better than physicians' judgment at predicting a man's risk of cancer. Without understanding the design and limitations of these methods, they can be applied inappropriately, leading to incorrect conclusions. These models are important components in counseling patients on their risk of prostate cancer and also help in the design of clinical trials by stratifying patients into different risk categories. Thus, it is incumbent on both clinicians and researchers to become familiar with these tools. Cancer 2009;115(13 suppl):3085 99. (c) 2009 American Cancer Society. PMID- 19544549 TI - Systems pathology: a paradigm shift in the practice of diagnostic and predictive pathology. AB - Diagnostic tumor pathology in the context of personalized medicine has progressed from an interpretive, subjective science to a more objective, evidence-based practice. This has resulted in the development of several tissue-based, molecular driven tests that provide information regarding prognosis and response to therapy. The challenge, however, for both the pathologist and the treating physician is how best to effectively integrate this data into a comprehensive treatment plan that includes a patient-specific risk assessment. To address this need, the authors developed a systems pathology approach to the practice of clinical molecular medicine through technical advances in object-oriented image analysis, and phenotyping at the microanatomical level using deparaffinized tissue section and quantitative biomarker multiplexing. With support vector regression for censored data, they have been able to integrate complex information and provide a patient-specific risk profile based on the clinical endpoint under investigation. Cancer 2009;115(13 suppl):3078-84. (c) 2009 American Cancer Society. PMID- 19544551 TI - Predictive models in diagnosing indolent cancer. AB - The early detection of prostate cancer has led to the increased incidence of tumors that are unlikely to become symptomatic during life: so-called indolent cancers. The ability to predict indolent prostate cancer is needed to avoid over treatment by unnecessary invasive therapies and to select men for active surveillance. For this report, the currently available nomograms that predict these low-risk tumors were reviewed. Many of these nomograms were based on clinical patient series, thus, their general application is restricted. Those nomograms based on screening series of the general population need further validation before generalized application is feasible. Cancer 2009;115(13 suppl):3100-6. (c) 2009 American Cancer Society. PMID- 19544552 TI - Racial and ethnic disparities in the incidence of invasive cervical cancer in Florida. AB - BACKGROUND: Although cervical cancer incidence has declined in the past decade, considerable racial and ethnic differences remain. The objective of this study was to examine differences in incidence by histology and cancer stage in Florida stratified further by race, ethnicity, and 5-year time intervals. METHODS: Women who were diagnosed with invasive cervical cancer in Florida between January 1985 and December 2004 were included in the analysis. Age-adjusted incidence rates by race and ethnicity were estimated for different histologic types and stages of cancer. The annual percentage of change in incidence also was calculated for each histologic type. Rate ratios were estimated by race and ethnicity using whites and non-Hispanics as the reference group. RESULTS: Overall, the incidence in Florida of cervical squamous cell carcinoma and transitional cell carcinoma declined significantly from 9.1 per 100,000 women in 1985 to 5.6 per 100,000 women in 2004 (P < .05), whereas the incidence of cervical adenocarcinoma remained stable (P > .05). The incidence of invasive cervical cancer was 9.6 per 100,000 women among whites and 13.13 per 100,000 women among African Americans from 2000 to 2004. African-American women were nearly 2 times more likely to be diagnosed at regional and distant cancer stages than white women for all periods examined. Furthermore, among African-American women aged >40 years, the age specific incidence of invasive cervical cancer increased considerably, whereas the rates among other racial groups decreased. CONCLUSIONS: The increasing rate of invasive cervical cancer among African-American women aged >40 years in Florida, coupled with their diagnosis at a later stage of cancer, is of great concern. Most screening organizations recommend stopping screening at age 65 years. The observations from these analyses highlighted the need to focus prevention and screening efforts on African-American women living in Florida, and particularly on women of postreproductive age. PMID- 19544553 TI - Predictive modeling in prostate cancer. Proceedings of the 2008 European School of Oncology Inside Track Conference. PMID- 19544554 TI - The 2008 European School of Oncology inside track conference, "predictive modeling in prostate cancer". AB - The European School of Oncology (ESO) Inside Track Conference, "Predictive Modeling in Prostate Cancer," the first event ever dedicated to prediction in prostate cancer, was organized in collaboration with the Prostate Program of Milan National Cancer Institute and the American Italian Cancer Foundation in the wonderful scenario of the Excelsior Lido Hotel in Venice on April 17 through 19, 2008. More than 240 participants from 23 countries attended this 3-day conference, which convened an exceptional group of experts from all over the world whose presentations provided a framework for understanding the state of the art in predictive modeling of prostate cancer and displayed future research trends in the uro-oncologic community. Cancer 2009;115(13 suppl):3035-8. (c) 2009 American Cancer Society. PMID- 19544555 TI - Anorexia nervosa: a role for L-arginine supplementation in cardiovascular risk factors? AB - OBJECTIVE: The aim of the present study was to evaluate the effects of a short term oral L-arginine treatment on platelet NO production, intracellular calcium concentration, iNOS and eNOS expression, in AN patients. METHOD: Forty outpatients belonging to restricting subtype and 40 normal participants age and sex matched were enrolled in the study. RESULTS: NO production was significantly elevated in the platelets from AN patients compared with controls while [Ca(2+)](i) was significantly decreased in patients with respect to controls. Western blot analysis demonstrated that iNOS isoform was more pronounced in the cell lysates from AN patients than controls. After supplementation with L arginine, both NO production and [Ca(2+)](i) seem to return to control levels, suggesting a probable recovery of their metabolisms. The same was found after western blot analysis of NOS expression. DISCUSSION: The results here proposed can be considered highly indicative of a positive effect of L-arginine supplementation on platelet NO production in AN patients. PMID- 19544556 TI - Therapeutic writing as an intervention for symptoms of bulimia nervosa: effects and mechanism of change. AB - OBJECTIVE: This study explored the effects on bulimic symptomatology of a writing task intended to reduce emotional avoidance. METHOD: Eighty individuals reporting symptoms of bulimia completed, by e-mail, a therapeutic or control writing task. Participants completed questionnaires on bulimic symptoms, mood, and potential moderating and mediating factors, and were followed up after 4 and 8 weeks. Writing content was explored using a word count package and qualitative framework analysis. RESULTS: Bulimic symptoms decreased in both groups, although in both groups the number of participants who improved was approximately equal to the number who did not improve. Symptom decreases were associated with increases in perceived mood regulation abilities, and decreases in negative beliefs about emotions. Participants preferred internet delivery to face to face discussion. DISCUSSION: For individuals experiencing symptoms of bulimia, the effects of therapeutic writing did not differ significantly from effects of a control writing task. PMID- 19544557 TI - A 4-year prospective study of eating disorder NOS compared with full eating disorder syndromes. AB - OBJECTIVE: To examine the course of Eating Disorder NOS (EDNOS) compared with anorexia nervosa (AN), bulimia nervosa (BN), and binge eating disorder (BED). METHOD: Prospective study of 385 participants meeting DSM-IV criteria for AN, BN, BED, and EDNOS at three sites. Recruitment was from the community and specialty clinics. Participants were followed at 6-month intervals during a 4-year period using the Eating Disorder Examination as the primary assessment. RESULTS: EDNOS remitted significantly more quickly that AN or BN but not BED. There were no differences between EDNOS and full ED syndromes, or the subtypes of EDNOS, in time to relapse following first remission. Only 18% of the EDNOS group had never had or did not develop another ED diagnosis during the study; however, this group did not differ from the remaining EDNOS group. DISCUSSION: EDNOS appears to be a way station between full ED syndromes and recovery, and to a lesser extent from recovery or EDNOS status to a full ED. Implications for DSM-V are examined. PMID- 19544558 TI - Mortality and predictors of death in a cohort of patients presenting to an eating disorders service. AB - OBJECTIVE: The main aim was to investigate mortality across the spectrum of eating disorders presenting to a specialized service for adults in the UK. A secondary aim was to explore whether any clinical and demographic factors may be associated with increased risk. METHOD: We conducted a "tracing" study of 1,892 patients assessed by the Service between 1992 and 2004. We used the Office of National Statistics to identify all deaths recorded up to August 2007. We also compared deceased patients with matched controls in terms of a number of clinical and demographic variables at initial presentation to the service. RESULTS: Anorexia nervosa had a ten-fold increased risk of early death, but there was also evidence of increased risk in other groups of patients, including eating disorder not otherwise specified. A number of clinical factors predicted increased mortality risk, especially very low body mass index and the presence of alcohol misuse. DISCUSSION: The study confirms previous evidence of a markedly increased mortality risk for anorexia nervosa, but also suggests that the risk is not confined to this eating disorder and that greater attention needs to be focused on wider psychiatric comorbidity. PMID- 19544559 TI - Synergistic effect of HLA class II loci and cytokine gene polymorphisms on the risk of gastric cancer in Japanese patients with Helicobacter pylori infection. AB - It has been reported that polymorphisms of human leukocyte antigen (HLA) genes and several cytokine genes are associated with an increased risk of developing gastric cancer (GC). However, the results of studies from different geographic regions, ethnic groups and study groups are inconsistent. The aim of this study was to evaluate the influence of H. pylori infection and host genetic factors on GC susceptibility in Japanese patients with GC. We analyzed genotypes for HLA class I and II, tumor necrosis factor alpha, interleukin (IL)-1beta, IL-1 receptor, IL-4, IL-4Ralpha and IL-10 in 330 H. pylori-infected noncardia patients with GC and 190 H. pylori-infected nonulcer dyspeptic controls. Haplotype analyses indicated that the frequencies of the HLA DRB1*0405 and DQB1*0401 alleles were increased in the patients with intestinal-type GC when compared with controls (both DRB1*0405 and DQB1*0401: p = 0.015, OR = 1.57, 95% CI = 1.09 2.26), but the changes were not statistically significant after correction for multiple comparisons. None of the cytokine gene polymorphisms were associated with GC susceptibility, whether patients with GC were analyzed as a group according to the histological subtype. Of interest was the comparison of controls and patients with intestinal-type GC. The frequency of an IL-10-592AA homozygote showing concomitant carriage of the HLA DRB1*0405-DQB1*0401 haplotype was significantly higher in patients with intestinal-type GC (chi(2) = 6.369, p = 0.0116, p(c) = 0.0464, OR = 2.43, 95% CI = 1.21-4.48). Our results suggest that the HLA class II and IL-10-592A/C polymorphisms synergistically affect the susceptibility to GC development of H. pylori-infected individuals in the Japanese population. PMID- 19544561 TI - Adiponectin is not effective against AOM-induced colon cancer but more evidence is required for its role in obesity-associated colon cancer: comment on the study by Ealey and Archer (2009). PMID- 19544563 TI - The novel HSP90 inhibitor STA-1474 exhibits biologic activity against osteosarcoma cell lines. AB - Osteosarcoma (OSA), the most common malignant bone tumor in dogs and children, exhibits a similar clinical presentation and molecular biology in both species. Unfortunately, 30-40% of children and 90% of dogs still die of disease despite aggressive therapy. The purpose of this study was to test the biologic activity of a novel heat shock protein 90 (HSP90) inhibitor, STA-1474, against OSA. Canine and human OSA cell lines and normal canine osteoblasts were treated with STA-1474 and evaluated for effects on proliferation (CyQuant), apoptosis (Annexin V, PARP cleavage, caspase 3/7 activation) and known HSP90 client proteins. HSP90 was immunoprecipitated from normal and malignant osteoblasts and Western blotting for co-chaperones was performed. Mice bearing canine OSA xenografts were treated with STA-1474, and tumors samples were evaluated for caspase-3 activation and loss of p-Akt/Akt. Treatment with STA-1474 promoted loss of cell viability, inhibition of cell proliferation and induction of apoptosis in OSA cell lines. STA-1474 and its active metabolite STA-9090 also demonstrated increased potency compared to 17 AAG. STA-1474 exhibited selectivity for OSA cells versus normal canine osteoblasts, and HSP90 co-precipitated with co-chaperones p23 and Hop in canine OSA cells but not in normal canine osteoblasts. Furthermore, STA-1474 downregulated the expression of p-Met/Met, p-Akt/Akt and p-STAT3. Finally, STA 1474 induced tumor regression, caspase-3 activation and downregulation of p Met/Met and p-Akt/Akt in OSA xenografts. Together, these data suggest that HSP90 represents a relevant target for therapeutic intervention in OSA. PMID- 19544560 TI - Antagonism of chemokine receptor CXCR3 inhibits osteosarcoma metastasis to lungs. AB - Metastasis continues to be the leading cause of mortality for patients with cancer. Several years ago, it became clear that chemokines and their receptors could control the tumor progress. CXCR3 has now been identified in many cancers including osteosarcoma and CXCR3 ligands were expressed by lungs that are the primary sites to which this tumor metastasize. This study tested the hypothesis that disruption of the CXCR3/CXCR3 ligands complexes could lead to a decrease in lungs metastasis. The experimental design involved the use of the CXCR3 antagonist, AMG487 and 2 murine models of osteosarcoma lung metastases. After tail vein injection of osteosarcoma cells, mice that were systematically treated with AMG487 according to preventive or curative protocols had a significant reduction in metastatic disease. Treatment of osteosarcoma cells in vitro with AMG487 led to decreased migration, decreased matrix metalloproteinase activity, decreased proliferation/survival and increased caspase-independent death. Taken together, our results support the hypothesis that CXCR3 and their ligands intervene in the initial dissemination of the osteosarcoma cells to the lungs and stimulate the growth and expansion of the metastatic foci in later stages. Moreover, these studies indicate that targeting CXCR3 may specifically inhibit tumor metastasis without adversely affecting antitumoral host response. PMID- 19544564 TI - Prediction of global and local quality of CASP8 models by MULTICOM series. AB - Evaluating the quality of protein structure models is important for selecting and using models. Here, we describe the MULTICOM series of model quality predictors which contains three predictors tested in the CASP8 experiments. We evaluated these predictors on 120 CASP8 targets. The average correlations between predicted and real GDT-TS scores of the two semi-clustering methods (MULTICOM and MULTICOM CLUSTER) and the one single-model ab initio method (MULTICOM-CMFR) are 0.90, 0.89, and 0.74, respectively; and their average difference (or GDT-TS loss) between the global GDT-TS scores of the top-ranked models and the best models are 0.05, 0.06, and 0.07, respectively. The average correlation between predicted and real local quality scores of the semi-clustering methods is above 0.64. Our results show that the novel semi-clustering approach that compares a model with top ranked reference models can improve initial quality scores generated by the ab initio method and a simple meta approach. PMID- 19544566 TI - Assessment of global and local model quality in CASP8 using Pcons and ProQ. AB - Model Quality Assessment Programs (MQAPs) are programs developed to rank protein models. These methods can be trained to predict the overall global quality of a model or what local regions in a model that are likely to be incorrect. In CASP8, we participated with two predictors that predict both global and local quality using either consensus information, Pcons, or purely structural information, ProQ. Consistently with results in previous CASPs, the best performance in CASP8 was obtained using the Pcons method. Furthermore, the results show that the modification introduced into Pcons for CASP8 improved the predictions against GDT_TS and now a correlation coefficient above 0.9 is achieved, whereas the correlation for ProQ is about 0.7. The correlation is better for the easier than for the harder targets, but it is not below 0.5 for a single target and below 0.7 only for three targets. The correlation coefficient for the best local quality MQAP is 0.68 showing that there is still clear room for improvement within this area. We also detect that Pcons still is not always able to identify the best model. However, we show that using a linear combination of Pcons and ProQ it is possible to select models that are better than the models from the best single server. In particular, the average quality over the hard targets increases by about 6% compared with using Pcons alone. PMID- 19544565 TI - Structural study of Carcinus maenas hemocyanin by native ESI-MS: interaction with L-lactate and divalent cations. AB - The interaction of L-lactate and divalent cations with Carcinus maenas hemocyanin has been probed by electrospray ionization mass spectrometry under conditions preserving noncovalent interactions (native ESI-MS). C. maenas native hemocyanin in the hemolymph occurs mainly as dodecamers and to a lesser extent as hexamers. A progressive acidification with formic acid after alkaline dissociation resulted in the preferential recruitment of the two lightest subunits into light dodecamers, a molecular complex absent from native hemolymph, in addition to regular dodecamers and hexamers. Addition of L-lactic acid also induced the recruitment of these subunits, even at alkaline pH. A dodecamer-specific subunit is needed to enable aggregation over the hexameric state. Experiments with EDTA suggested the existence of different binding sites and association constants for divalent cations within hexameric structures and at the interface between two hexamers. L-lactic acid specific interaction with the lightest subunits was not inhibited by removal of the divalent cations. PMID- 19544567 TI - Insight into the substrate length restriction of M32 carboxypeptidases: characterization of two distinct subfamilies. AB - M32 carboxypeptidases are a distinct family of HEXXH metalloproteases whose structures exhibit a narrow substrate groove that is blocked at one end. Structural alignments with other HEXXH metalloprotease-peptide complexes suggested an orientation in which the substrate is directed towards the back of the groove. This led us to hypothesize, and subsequently confirm that the maximum substrate length for M32 carboxypeptidases is restricted. Structural and sequence analyses implicate a highly conserved Arg at the back of the groove as being critical for this length restriction. However, the Thermus thermophilus and Bacillus subtilis M32 members lack this conserved Arg. Herein, we present the biochemical and structural characterization of these two proteins. Our findings support the important role of the conserved Arg in maintaining the length restriction, and reveal a proline-rich loop as an alternate blocking strategy. Based on our results, we propose that M32 carboxypeptidases from Bacilli belong to a separate subfamily. PMID- 19544568 TI - X-ray crystallographic studies of RNase A variants engineered at the most destabilizing positions of the main hydrophobic core: further insight into protein stability. AB - To investigate the structural origin of decreased pressure and temperature stability, the crystal structure of bovine pancreatic ribonuclease A variants V47A, V54A, V57A, I81A, I106A, and V108A was solved at 1.4-2.0 A resolution and compared with the structure of wild-type protein. The introduced mutations had only minor influence on the global structure of ribonuclease A. The structural changes had individual character that depends on the localization of mutated residue, however, they seemed to expand from mutation site to the rest of the structure. Several different parameters have been evaluated to find correlation with decrease of free energy of unfolding DeltaDeltaG(T), and the most significant correlation was found for main cavity volume change. Analysis of the difference distance matrices revealed that the ribonuclease A molecule is organized into five relatively rigid subdomains with individual response to mutation. This behavior consistent with results of unfolding experiments is an intrinsic feature of ribonuclease A that might be surviving remnants of folding intermediates and reflects the dynamic nature of the molecule. PMID- 19544570 TI - Cohesin diversity revealed by the crystal structure of the anchoring cohesin from Ruminococcus flavefaciens. AB - The cellulosome is an intriguing multienzyme complex found in cellulolytic bacteria that plays a key role in the degradation of plant cell-wall polysaccharides. In Ruminococcus flavefaciens, a predominant fiber-degrading bacterium found in ruminants, the cellulosome is anchored to the bacterial cell wall through a relatively short ScaE scaffoldin. Determination of the crystal structure of the lone type-III ScaE cohesin from R. flavefaciens (Rf-CohE) was initiated as a part of a structural effort to define cellulosome assembly. The structure was determined at 1.95 A resolution by single-wavelength anomalous diffraction. This is the first detailed description of a crystal structure for a type-III cohesin, and its features were compared with those of the known type-I and type-II cohesin structures. The Rf-CohE module folds into a nine-stranded beta-sandwich with jellyroll topology, typically observed for cohesins, and includes two beta-flaps in the midst of beta-strands 4 and 8, similar to the type II cohesin structures. However, the presence in Rf-CohE of an additional 13 residue alpha-helix located between beta-strands 8 and 9 represents a dramatic divergence from other known cohesin structures. The prominent alpha-helix is enveloped by an extensive N-terminal loop, not observed in any other known cohesin, which embraces the helix presumably enhancing its stability. A planar surface at the upper portion of the front face of the molecule, bordered by beta flap 8, exhibits plausible dimensions and exposed amino acid residues to accommodate the dockerin-binding site. PMID- 19544569 TI - The 2.1 A crystal structure of an acyl-CoA synthetase from Methanosarcina acetivorans reveals an alternate acyl-binding pocket for small branched acyl substrates. AB - The acyl-AMP forming family of adenylating enzymes catalyze two-step reactions to activate a carboxylate with the chemical energy derived from ATP hydrolysis. X ray crystal structures have been determined for multiple members of this family and, together with biochemical studies, provide insights into the active site and catalytic mechanisms used by these enzymes. These studies have shown that the enzymes use a domain rotation of 140 degrees to reconfigure a single active site to catalyze the two partial reactions. We present here the crystal structure of a new medium chain acyl-CoA synthetase from Methanosarcina acetivorans. The binding pocket for the three substrates is analyzed, with many conserved residues present in the AMP binding pocket. The CoA binding pocket is compared to the pockets of both acetyl-CoA synthetase and 4-chlorobenzoate:CoA ligase. Most interestingly, the acyl-binding pocket of the new structure is compared with other acyl- and aryl-CoA synthetases. A comparison of the acyl-binding pocket of the acyl-CoA synthetase from M. acetivorans with other structures identifies a shallow pocket that is used to bind the medium chain carboxylates. These insights emphasize the high sequence and structural diversity among this family in the area of the acyl binding pocket. PMID- 19544571 TI - Crystal structure and enhanced activity of a cutinase-like enzyme from Cryptococcus sp. strain S-2. AB - The structural and enzymatic characteristics of a cutinase-like enzyme (CLE) from Cryptococcus sp. strain S-2, which exhibits remote homology to a lipolytic enzyme and a cutinase from the fungus Fusarium solani (FS cutinase), were compared to investigate the unique substrate specificity of CLE. The crystal structure of CLE was solved to a 1.05 A resolution. Moreover, hydrolysis assays demonstrated the broad specificity of CLE for short and long-chain substrates, as well as the preferred specificity of FS cutinase for short-chain substrates. In addition, site-directed mutagenesis was performed to increase the hydrolysis activity on long-chain substrates, indicating that the hydrophobic aromatic residues are important for the specificity to the long-chain substrate. These results indicate that hydrophobic residues, especially the aromatic ones exposed to solvent, are important for retaining lipase activity. PMID- 19544573 TI - Structure, dynamics, and interactions of jacalin. Insights from molecular dynamics simulations examined in conjunction with results of X-ray studies. AB - Molecular dynamics simulations have been carried out on all the jacalin carbohydrate complexes of known structure, models of unliganded molecules derived from the complexes and also models of relevant complexes where X-ray structures are not available. Results of the simulations and the available crystal structures involving jacalin permit delineation of the relatively rigid and flexible regions of the molecule and the dynamical variability of the hydrogen bonds involved in stabilizing the structure. Local flexibility appears to be related to solvent accessibility. Hydrogen bonds involving side chains and water bridges involving buried water molecules appear to be important in the stabilization of loop structures. The lectin-carbohydrate interactions observed in crystal structures, the average parameters pertaining to them derived from simulations, energetic contribution of the stacking residue estimated from quantum mechanical calculations, and the scatter of the locations of carbohydrate and carbohydrate-binding residues are consistent with the known thermodynamic parameters of jacalin-carbohydrate interactions. The simulations, along with X ray results, provide a fuller picture of carbohydrate binding by jacalin than provided by crystallographic analysis alone. The simulations confirm that in the unliganded structures water molecules tend to occupy the positions occupied by carbohydrate oxygens in the lectin-carbohydrate complexes. Population distributions in simulations of the free lectin, the ligands, and the complexes indicate a combination of conformational selection and induced fit. PMID- 19544572 TI - Crystal structure of a histidine kinase sensor domain with similarity to periplasmic binding proteins. PMID- 19544574 TI - Comparative 3D quantitative analyses of trapeziometacarpal joint surface curvatures among living catarrhines and fossil hominins. AB - Comparisons of joint surface curvature at the base of the thumb have long been made to discern differences among living and fossil primates in functional capabilities of the hand. However, the complex shape of this joint makes it difficult to quantify differences among taxa. The purpose of this study is to determine whether significant differences in curvature exist among selected catarrhine genera and to compare these genera with hominin fossils in trapeziometacarpal curvature. Two 3D approaches are used to quantify curvatures of the trapezial and metacarpal joint surfaces: (1) stereophotogrammetry with nonuniform rational B-spline (NURBS) calculation of joint curvature to compare modern humans with captive chimpanzees and (2) laser scanning with a quadric based calculation of curvature to compare modern humans and wild-caught Pan, Gorilla, Pongo, and Papio. Both approaches show that Homo has significantly lower curvature of the joint surfaces than does Pan. The second approach shows that Gorilla has significantly more curvature than modern humans, while Pongo overlaps with humans and African apes. The surfaces in Papio are more cylindrical and flatter than in Homo. Australopithecus afarensis resembles African apes more than modern humans in curvatures, whereas the Homo habilis trapezial metacarpal surface is flatter than in all genera except Papio. Neandertals fall at one end of the modern human range of variation, with smaller dorsovolar curvature. Modern human topography appears to be derived relative to great apes and Australopithecus and contributes to the distinctive human morphology that facilitates forceful precision and power gripping, fundamental to human manipulative activities. PMID- 19544575 TI - Links between habitat degradation, and social group size, ranging, fecundity, and parasite prevalence in the Tana River mangabey (Cercocebus galeritus). AB - We investigated the effects of anthropogenic habitat degradation on group size, ranging, fecundity, and parasite dynamics in four groups of the Tana River mangabey (Cercocebus galeritus). Two groups occupied a forest disturbed by human activities, while the other two occupied a forest with no human disturbance. We predicted that the groups in the disturbed forest would be smaller, travel longer distances daily, and have larger home ranges due to low food tree abundance. Consequently, these groups would have lower fecundity and higher parasite prevalence and richness (number of parasite species). We measured the abundance of food trees and anthropogenic activity in the forests, the groups' daily travel distances and home range sizes, and censused social groups over 12 months. We also analyzed fecal samples for gastrointestinal parasites from three of the groups. The disturbed forest had a lower abundance of food trees, and groups in this forest traveled longer distances, had larger home range sizes, were smaller, and had lower fecundity. The groups in the disturbed forest had higher, although not statistically significant, parasite prevalence and richness. This study contributes to a better understanding of how anthropogenic habitat change influences fecundity and parasite infections in primates. Our results also emphasize the strong influence of habitat quality in determining daily travel distance and home range size in primates. PMID- 19544576 TI - Trophy-taking and dismemberment as warfare strategies in prehistoric central California. AB - We document evidence for trophy-taking and dismemberment with a new bioarchaeological database featuring 13,453 individuals from prehistoric central California sites. Our study reveals 76 individuals with perimortem removal of body parts consistent with trophy-taking or dismemberment; nine of these individuals display multiple types of trophy-taking and dismemberment for a total of 87 cases. Cases span almost 5,000 years, from the Early Period (3000-500 BC) to the Late Period (AD 900-1700). Collectively, these individuals share traits that distinguish them from the rest of the population: a high frequency of young adult males, an increased frequency of associated trauma, and a tendency towards multiple burials and haphazard burial positions. Eight examples of human bone artifacts were also found that appear related to trophy-taking. These characteristics suggest that trophy-taking and dismemberment were an important part of the warfare practices of central Californian tribes. Temporally, the two practices soared in the Early/Middle Transition Period (500-200 BC), which may have reflected a more complex sociopolitical system that encouraged the use of trophies for status acquisition, as well as the migration of outside groups that resulted in intensified conflict. Overall, trophy-taking and dismemberment appear to have been the product of the social geography of prehistoric central California, where culturally differentiated tribes lived in close proximity to their enemies. PMID- 19544577 TI - A new species of Pliopithecus Gervais, 1849 (Primates: Pliopithecidae) from the Middle Miocene (MN8) of Abocador de Can Mata (els Hostalets de Pierola, Catalonia, Spain). AB - Pliopithecus (Pliopithecus) canmatensis sp. nov. is described from several Late Aragonian localities from Abocador de Can Mata (ACM) in els Hostalets de Pierola (Valles-Penedes Basin, Catalonia, Spain), spanning from approximately 11.7 to 11.6 Ma (C5r.3r subchron), and being correlated to the MN8 (reference locality La Grive L3). The ACM remains display a pliopithecine dental morphology with well developed pliopithecine triangles on M/2 and M/3. This, together with other occlusal details, negates an attribution to the subgenus Epipliopithecus. Although slightly smaller, the ACM remains are most similar in size to comparable elements of P. piveteaui and P. antiquus. Several occlusal details (such as the greater development of the buccal cingulid in lower molars) and dental proportions (M/3 much longer than M/2), however, indicate greater similarities with P. antiquus from Sansan and La Grive. The ACM remains, however, differ from P. antiquus in dental proportions as well as occlusal morphology of the lower molars (including the less peripheral position of the protoconid and more medial position of the hypoconulid, the more mesial position of the buccal cuspids as compared to the lingual ones, the narrower but distinct mesial fovea, the higher trigonid, and the more extensive buccal cingulid, among others). These differences justify a taxonomic distinction at the species level of the ACM pliopithecid remains with respect to P. antiquus. Previous pliopithecid findings from the Valles-Penedes Basin, previously attributed to P. antiquus, are neither attributable to the latter species nor to the newly erected one. PMID- 19544578 TI - Cofactor binding and enzymatic activity in an unevolved superfamily of de novo designed 4-helix bundle proteins. AB - To probe the potential for enzymatic activity in unevolved amino acid sequence space, we created a combinatorial library of de novo 4-helix bundle proteins. This collection of novel proteins can be considered an "artificial superfamily" of helical bundles. The superfamily of 102-residue proteins was designed using binary patterning of polar and nonpolar residues, and expressed in Escherichia coli from a library of synthetic genes. Sequences from the library were screened for a range of biological functions including heme binding and peroxidase, esterase, and lipase activities. Proteins exhibiting these functions were purified and characterized biochemically. The majority of de novo proteins from this superfamily bound the heme cofactor, and a sizable fraction of the proteins showed activity significantly above background for at least one of the tested enzymatic activities. Moreover, several of the designed 4-helix bundles proteins showed activity in all of the assays, thereby demonstrating the functional promiscuity of unevolved proteins. These studies reveal that de novo proteins which have neither been designed for function, nor subjected to evolutionary pressure (either in vivo or in vitro)-can provide rudimentary activities and serve as a "feedstock" for evolution. PMID- 19544579 TI - Cystein-serine-rich nuclear protein 1, Axud1/Csrnp1, is essential for cephalic neural progenitor proliferation and survival in zebrafish. AB - The CSRNP (cystein-serine-rich nuclear protein) family has been conserved from Drosophila to human. Although knockout mice for each of the mammalian proteins have been generated, their function during vertebrate development has remained elusive. As an alternative to obtain insights on CSRNP's role in development, we have analysed the expression pattern and function of one member of this family, axud1, during zebrafish development. Our expression analysis indicates that axud1 is expressed from cleavage to larval stages in a dynamic pattern, becoming restricted after gastrulation to anterior regions of the developing neuraxis and later on concentrated predominantly in proliferating domains of the brain. Knockdown analysis using antisense morpholinos shows that reducing Axud1 levels impairs neural progenitor cell proliferation and survival, revealing an essential function of this gene for the growth of cephalic derivatives. The brain growth phenotypes elicited by decreasing Axud1 expression are specific and independent of anterior-posterior patterning events, initial establishment of neural progenitors, or neural differentiation occurring in this tissue. However, Axud1 is necessary for six3.1 expression and is positively regulated by sonic hedgehog. Phylogenetic examination shows that axud1 is likely to be the ortholog of the only member of this family present in Drosophila, as well as to the previously described mouse CSRNP1 and to human AXUD1 (Axin upregulated-1). Thus, we provide evidence as to the role of axud1 in brain growth in vertebrates. PMID- 19544580 TI - Effect of protein structure on deamidation rate in the Fc fragment of an IgG1 monoclonal antibody. AB - The effects of secondary structure on asparagine (N) deamidation in a 22 amino acid sequence (369-GFYPSDIAVEWESNGQPENNYK-390) of the crystallizable (Fc) fragment of a human monoclonal antibody (Fc IgG1) were investigated using high resolution ultra performance liquid chromatography with tandem mass spectrometry (UPLC/MS). Samples containing either the intact Fc IgG (approximately 50 kD) ("intact protein"), or corresponding synthetic peptides ("peptide") were stored in Tris buffer at 37 degrees C and pH 7.5 for up to forty days, then subjected to UPLC/MS analysis with high energy MS1 fragmentation. The peptide deamidated only at N(382) to form the isoaspartate (isoD(382)) and aspartate (D(382)) products in the ratio of approximately 4:1, with a half-life of approximately 3.4 days. The succinimide intermediate (Su(382)) was also detected; deamidation was not observed for the other two sites (N(387) and N(388)) in peptide samples. The intact protein showed a 30-fold slower overall deamidation half-life of approximately 108 days to produce the isoD(382) and D(387) products, together with minor amounts of D(382). Surprisingly, the D(382) and isoD(387) products were not detected in intact protein samples and, as in the peptide samples, deamidation was not detected at N(388). The results indicate that higher order structure influences both the rate of N-deamidation and the product distribution. PMID- 19544581 TI - Vibrational spectroscopic investigation of the ligand binding domain of kainate receptors. AB - Fourier transform infrared spectroscopy has been used to probe the agonist protein interactions in the ligand binding domain of the GluR6 subunit, one subunit of the kainate subtype of glutamate receptors. In order to study the changes in the interactions over a range of activations the investigations were performed using the wild type, N690S, and T661E mutations. These studies show that the strength of the interactions at the alpha-amine group of the agonist, as probed by studying the environment of the nondisulphide bonded Cys 432, acts as a switch with weaker interactions at lower activations and stronger interactions at higher activations. The alpha-carboxylate interactions of the agonist, however, are not significantly different over the wide range of activations, as measured by the maximum currents mediated by the receptors at saturating concentrations of agonists. Previous investigations of AMPA receptors show a similar dependence of the alpha-amine interactions on activation indicating that the roles of the alpha amine interactions in mediating receptor activation are similar for both subtypes of receptors; however, in the case of the AMPA receptors a tug of war type of change was observed between the alpha-amine and alpha-carboxylate interactions and this is not observed in kainate receptors. This decoupling of the two interactions could arise due to the larger cleft observed in kainate receptors, which allows for a more flexible interaction for the alpha-amine and alpha carboxylate groups of the agonists. PMID- 19544582 TI - Fibroblast growth factor (FGF) gene expression in the developing cerebellum suggests multiple roles for FGF signaling during cerebellar morphogenesis and development. AB - The cerebellum is derived from the anterior-most segment of the embryonic hindbrain, rhombomere 1 (r1). Previous studies have shown that the early development and patterning of r1 requires fibroblast growth factor (FGF) signaling. However, many of the developmental processes that shape cerebellar morphogenesis take place later in embryonic development and during the first 2 weeks of postnatal life in the mouse. Here, we present a more comprehensive analysis of the expression patterns of genes encoding FGF receptors and secreted FGF ligands during these later stages of cerebellar development. We show that these genes are expressed in multiple cell types in the developing cerebellum, in an astonishing array of distinct patterns. These data suggest that FGF signaling functions throughout cerebellar development to regulate many processes that shape the formation of a functional cerebellum. PMID- 19544583 TI - Effects of activation of hedgehog signaling on patterning, growth, and differentiation in Xenopus froglet limb regeneration. AB - Regenerating limbs of urodele amphibians and Xenopus tadpole are reconstructed along proximal-distal, anterior-posterior (AP), and dorsal-ventral axes. In contrast, a regenerated limb of the Xenopus froglet does not have digits, and only a simple cartilaginous structure referred to as a "spike" is formed. This suggests that repatterning along the AP axis is absent in the froglet blastema. Previous studies have shown that Shh and its target genes are not expressed in the froglet blastema. In this study, we activated Hedgehog signaling in the froglet blastema and found that target genes of Shh were inducible in the mesenchyme of limb blastema. Furthermore, we found that activation of the signaling had effects on blastema cell proliferation and chondrogenesis and resulted in the formation of multiple cartilaginous structures. These findings indicate that activation of signaling that is absent in the froglet blastema is effective for improvement of limb regeneration ability in the Xenopus froglet. PMID- 19544585 TI - Crosstalk between Wnt and bone morphogenic protein signaling: a turbulent relationship. AB - The Wnt and the bone morphogenic protein (BMP) pathways are evolutionarily conserved and essentially independent signaling mechanisms, which, however, often regulate similar biological processes. Wnt and BMP signaling are functionally integrated in many biological processes, such as embryonic patterning in Drosophila and vertebrates, formation of kidney, limb, teeth and bones, maintenance of stem cells, and cancer progression. Detailed inspection of regulation in these and other tissues reveals that Wnt and BMP signaling are functionally integrated in four fundamentally different ways. The molecular mechanism evolved to mediate this integration can also be summarized in four different ways. However, a fundamental aspect of functional and mechanistic interaction between these pathways relies on tissue-specific mechanisms, which are often not conserved and cannot be extrapolated to other tissues. Integration of the two pathways contributes toward the sophisticated means necessary for creating the complexity of our bodies and the reliable and healthy function of its tissues and organs. PMID- 19544586 TI - In vitro and in vivo differentiation of human embryonic stem cells into retina like organs and comparison with that from mouse pluripotent epiblast stem cells. AB - Correctly inducing the differentiation of pluripotent hESCs to a specific lineage with high purity is highly desirable for regenerative cell therapy. Our first effort to perform in vitro differentiation of hESCs resulted in a limited recapitulation of the ocular tissue structures. When undifferentiated hESCs were placed in vivo into the ocular tissue, in this case into the vitreous cavity, 3 dimensional retina-like structures reminiscent of the invagination of the optic vesicle were generated. Immunohistochemical analysis confirmed the presence of both a neural retina-like cell layer and a retinal pigmented epithelium-like cell layer, possibly equivalent to the developing E12.5 mouse retina. Furthermore, mouse epiblast-derived stem cells, which are reported to share some characteristics with hESCs, but not with mouse ESCs, also generated retinal anlage-like structures in vivo. hESC-derived retina-like structures present a novel therapeutic possibility for retinal diseases and also provide a novel experimental system to study early human eye development. PMID- 19544584 TI - Paramagnetic relaxation enhancements in unfolded proteins: theory and application to drkN SH3 domain. AB - Site-directed spin labeling in combination with paramagnetic relaxation enhancement (PRE) measurements is one of the most promising techniques for studying unfolded proteins. Since the pioneering work of Gillespie and Shortle (J Mol Biol 1997;268:158), PRE data from unfolded proteins have been interpreted using the theory that was originally developed for rotational spin relaxation. At the same time, it can be readily recognized that the relative motion of the paramagnetic tag attached to the peptide chain and the reporter spin such as (1)H(N) is best described as a translation. With this notion in mind, we developed a number of models for the PRE effect in unfolded proteins: (i) mutual diffusion of the two tethered spheres, (ii) mutual diffusion of the two tethered spheres subject to a harmonic potential, (iii) mutual diffusion of the two tethered spheres subject to a simulated mean-force potential (Smoluchowski equation); (iv) explicit-atom molecular dynamics simulation. The new models were used to predict the dependences of the PRE rates on the (1)H(N) residue number and static magnetic field strength; the results are appreciably different from the Gillespie-Shortle model. At the same time, the Gillespie-Shortle approach is expected to be generally adequate if the goal is to reconstruct the distance distributions between (1)H(N) spins and the paramagnetic center (provided that the characteristic correlation time is known with a reasonable accuracy). The theory has been tested by measuring the PRE rates in three spin-labeled mutants of the drkN SH3 domain in 2M guanidinium chloride. Two modifications introduced into the measurement scheme-using a reference compound to calibrate the signals from the two samples (oxidized and reduced) and using peak volumes instead of intensities to determine the PRE rates-lead to a substantial improvement in the quality of data. The PRE data from the denatured drkN SH3 are mostly consistent with the model of moderately expanded random-coil protein, although part of the data point toward a more compact structure (local hydrophobic cluster). At the same time, the radius of gyration reported by Choy et al. (J Mol Biol 2002;316:101) suggests that the protein is highly expanded. This seemingly contradictory evidence can be reconciled if one assumes that denatured drkN SH3 forms a conformational ensemble that is dominated by extended conformations, yet also contains compact (collapsed) species. Such behavior is apparently more complex than predicted by the model of a random-coil protein in good solvent/poor solvent. PMID- 19544587 TI - Smoking induced worsening of dystonia. A case report. PMID- 19544589 TI - A novel approach to the rapid assignment of (13)C NMR spectra of major components of vegetable oils such as avocado, mango kernel and macadamia nut oils. AB - Assignment of (13)C nuclear magnetic resonance (NMR) spectra of major fatty acid components of South African produced vegetable oils was attempted using a method in which the vegetable oil was spiked with a standard triacylglycerol. This proved to be inadequate and therefore a new rapid and potentially generic graphical linear correlation method is proposed for assignment of the (13)C NMR spectra of major fatty acid components of apricot kernel, avocado pear, grapeseed, macadamia nut, mango kernel and marula vegetable oils. In this graphical correlation method, chemical shifts of fatty acids present in a known standard triacylglycerol is plotted against the corresponding chemical shifts of fatty acids present in the vegetable oils. This new approach (under carefully defined conditions and concentrations) was found especially useful for spectrally crowded regions where significant peak overlap occurs and was validated with the well-known (13)C NMR spectrum of olive oil which has been extensively reported in the literature. In this way, a full assignment of the (13)C{1H} NMR spectra of the vegetable oils, as well as tripalmitolein was readily achieved and the resonances belonging to the palmitoleic acid component of the triacylglycerols in the case of macadamia nut and avocado pear oil resonances were also assigned for the first time in the (13)C NMR spectra of these oils. PMID- 19544590 TI - Current aspects of aqueous humor dynamics and glaucoma - A Tribute to Douglas H. Johnson. PMID- 19544591 TI - Proceedings and abstracts of the 16th International Symposium on Endoscopic Ultrasonography, September 12-13, 2008, San Francisco, California, USA. PMID- 19544588 TI - Cognitive profiles of Chinese adolescents with dyslexia. AB - The present study sought to identify cognitive abilities that might distinguish Hong Kong Chinese adolescents with and without dyslexia and examined the cognitive profile of dyslexic adolescents in order to better understand this important problem. The performance of 27 Chinese adolescents with childhood diagnoses of dyslexia was compared with 27 adolescents of the same chronological age (CA) and 27 of matched reading level (RL) on measures of literacy and cognitive abilities: Chinese word reading, one-minute reading, reading comprehension, dictation, verbal short-term memory, rapid naming, visual orthographic knowledge, morphological and phonological awareness. The results indicated that the dyslexic group scored lower than the CA group, but similar to the RL group, especially in the areas of rapid naming, visual-orthographic knowledge and morphological awareness, with over half having multiple deficits exhibited 2 or more cognitive areas. Furthermore, the number of cognitive deficits was associated with the degree of reading and spelling impairment. These findings suggest that adolescents with childhood diagnoses of dyslexia have persistent literacy difficulties and seem to have multiple causes for reading difficulties in Chinese. PMID- 19544592 TI - Special issue: Future Directions in Cannabinoid Therapeutics II: From the Bench to the Clinic. Proceedings of the Center for Medicinal Cannabis Research (CMCR) June 2004 Workshop. PMID- 19544593 TI - Climate talks snarled up. PMID- 19544594 TI - Seroprevalence of HTLV-1/2 infection among native and immigrant pregnant women in Spain. AB - HTLV-1=2 antenatal screening is not mandatory in European countries. The rapid increase in immigrants coming from areas endemic for HTLV-1 infection has compelled a review of this policy in Spain. From February 2006 to December 2007, a cross-sectional study was carried out in all pregnant women attended at 10 different Spanish hospitals. An enzyme immunoassay (EIA) was used to test serum HTLV-1=2 antibodies; reactive samples were further confirmed by Western blot and=or polymerase chain reaction. A total of 20,518 pregnant women were examined, of whom 18,266 (89%) were native Spaniards. Overall, 946 (4.6%) of the immigrants came from HTLV-1 endemic areas (mainly Central and South America and sub-Saharan Africa). Four samples were EIA seroreactive for HTLV-1=2, two of them in women infected with HTLV-1 coming from endemic areas. The other two women were infected with HTLV-2; one was an immigrant from Bolivia and another was a native Spaniard who admitted prior injection drug use and was HIV-1 positive. The overall HTLV 1=2 seroprevalence was 0.19 per 1000 (95% CI: 0.05-0.49=1000). For HTLV-1, the seroprevalence was 2.11 per 1000 (95% CI: 0.26-7.62=1000) in pregnant women from endemic areas. The seroprevalence of HTLV-1=2 infection is below 0.02% among pregnant women in Spain, and therefore universal screening for HTLV-1=2 infection in antenatal clinics is not warranted. However, HTLV-1=2 screening could be considered in pregnant women coming from endemic areas, in whom the rate of infection is nearly 1000-fold higher than in native Spaniards and are the only group infected with the more pathogenic HTLV-1. PMID- 19544596 TI - Wanted, dead or alive. PMID- 19544595 TI - From the editors: series of articles on modelling. PMID- 19544597 TI - Phylogenomics and its Future: Devoted to Masami Hasegawa. PMID- 19544599 TI - Proceedings of the 5th International Conference on Countercurrent Chromatography Dedicated to Dr. Yoichiro Ito on the Occasion of his 80th Birthday, Rio de Janeiro, Brazil, 26-29 July 2008. PMID- 19544598 TI - Studies on the Ocular Lens - Dedicated to George Duncan. PMID- 19544600 TI - Special issue: Biomarker discovery and related topics, dedicated to the 70th birthday of Professor Barry Karger. PMID- 19544601 TI - Patchy pig monitoring may hide flu threat. PMID- 19544602 TI - Commentary on DuBois. PMID- 19544603 TI - Comment on DuBois's article, "Increasing rates of organ donation: exploring the IOM's boldest recommendation". PMID- 19544604 TI - HIV clinicians and patients should prepare now for fall pandemic flu. Recent novel H1N1 flu outbreak offers lessons. PMID- 19544605 TI - Recent flu vaccine and HIV trial holds promising news. HIV patients could quickly, safely enroll. PMID- 19544607 TI - HIV prevention trials require sophisticated community preparation. Controversial PrEP trials serve as cautionary tale. PMID- 19544606 TI - Q&A on H1N1 for people with HIV/AIDS. Drug-drug reactions not clear, should be reported. PMID- 19544608 TI - FDA notifications. EUA issued for treating novel 2009 H1N1 flu. PMID- 19544609 TI - FDA notifications. Tenofovir in India receives tentative approval. PMID- 19544610 TI - CDC launches new AIDS awareness campaign. PMID- 19544611 TI - 2009 H1N1/swine flu--anatomy of an outbreak. PMID- 19544612 TI - Swine-origin influenza A (H1N1) and HIV. The CDC offers guidance for HIV-infected patients potentially exposed to swine flu. PMID- 19544613 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. No clinical benefit from adding IL-2 to ART. PMID- 19544614 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. Timing of ART initiation--still no definitive answers. PMID- 19544615 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. Concomitant HIV/TB treatment improves survival. PMID- 19544616 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. Treatment is preventive, but some risk remains. PMID- 19544618 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. First (sort of) positive anti-HIV microbicide trial. PMID- 19544617 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. Lopinavir/r is superior to nevirapine in women who previously received single dose nevirapine. PMID- 19544619 TI - Assessment of lumbar spinal canal morphology with digital computed tomography. AB - BACKGROUND: The study of lumbar spine pathology requires adequate preparation and knowledge of the normal structure of this part of the spine. The main goal of the study was to assess spinal canal morphology with computed tomography. The sagittal and interpedicular dimensions and surface area were considered the most important measurements. An additional goal was to assess the shape of the spinal canal and intervertebral joint angles. MATERIAL AND METHODS: Computed tomography was used to assess the lumbar spinal canals of 42 people to an accuracy of 0.01 using a special console for digital analysis. RESULTS: The mean sagittal dimension showed minor differences and ranged from 15.75+/-0.886 at the L3 level to 17.77+/-1.619 at the L5 level. The mean interpedicular dimension was significantly different between the levels, increasing from 24.75+/-2.173 at L3 to 34.57+/-3.332 at L5. Similar results were obtained as regards the mean surface area of the spinal canal. The surface area was 277.2+/-36.15 mm2 at the L3 level, compared to 297+/-9.90 mm2 at L4 and 386.5+/-50.55 mm2 at L5. The spinal canal shape at the L4-L5 level was triangular or trefoil in all 42 patients. No significant differences were found between the angles of right and left intervertebral joints. CONCLUSIONS: Our results do not differ from those described in literature. A sagittal dimension over 15 mm, and an interpedicular dimension of more than 25 mm are characteristic of a normal spinal canal. The results constitute reference data for further studies concerning lumbar stenosis. PMID- 19544620 TI - [The message of the president of the IASGO-Romanian Section (International Association of Surgeons, Gastroenterologists and Oncologists) and of the ARCHBPTH (Romanian Association of Hepato-Bilio-Pancreatic Surgery and Liver Transplantation). The 11th Symposium of IASGO and the 4th Symposium of the ARCHBPTH]. PMID- 19544621 TI - Mitochondrial translocation of amyloid precursor protein and its cleaved products: relevance to mitochondrial dysfunction in Alzheimer's disease. AB - Alzheimer's disease (AD) is a multifactorial disorder. Mitochondrial dysfunction is one of the key characteristics of AD pathogenesis. However, the mechanisms underlying the progression of mitochondrial impairment during AD are not clear. Growing evidence suggests a causative role for intracellular accumulation of amyloid precursor protein (APP) and its proteolytic products in the pathogenesis of AD. Furthermore, APP possesses several domains including a mitochondrial targeting sequence. Recent literature suggests that mitochondrial localization of full length APP and its C-terminal proteolytically cleaved derivative beta amyloid (Abeta) are associated with the mitochondrial dysfunction. Here, we review the nature of mitochondrial localization of APP and Abeta and their pathological implications in AD mitochondrial dysfunction. PMID- 19544622 TI - A role for the human amygdala in higher cognition. AB - Considerable evidence suggests that the human amygdala plays an important role in higher cognitive functions in addition to its well-known role in emotional processing. In this article we review representative evidence showing the involvement of the human amygdala in long-term memory, working memory and attention. Results are discussed in terms of their relevance to current theories of amygdala function that can integrate its cognitive and emotional functions in a comprehensive framework. PMID- 19544623 TI - Reconsolidation revisited: a review and commentary on the phenomenon. AB - Consolidation and reconsolidation constitute a large proportion of current research into memory formation. The evidence in favour of the Consolidation Theory is widespread, on both the cellular and systems level. Research has indicated that consolidation and reconsolidation employ similar mechanisms; both consolidation and reconsolidation of memory require protein synthesis and glutaminergic input, and both seem to be associated with the hippocampal formation. Despite this, other data seem to argue that the two concepts are entirely separate processes. The great interest in this topic is shown in the proliferation of studies. The current literature has been subject to extensive and continual review. The current manuscript attempts to address the inconsistency in the consolidation-reconsolidation literature by providing a selective review of some of the most pertinent experimental work in both areas. The core question underpinning this review paper is whether reconsolidation is an entity distinct from consolidation, or merely an extension of the consolidation process. It is concluded that consolidation and reconsolidation may be distinct, albeit similar, processes, and that only a subset of the brain areas involved in consolidation are implicated in reconsolidation. In addition, with advances in our understanding of, and approach to these processes (i.e., incorporation of boundary conditions of reconsolidation into the design of contemporary studies and the increased awareness of the need to temper the interpretation of data emerging from studies employing divergent methodologies), it is suggested that future reconsolidation research may yield significant progress into the vast potential underpinning the reconsolidation phenomenon. PMID- 19544624 TI - Interaction between orexins and the mesolimbic system for overriding satiety. AB - In North American society, it is all too common for the intake of calories to outweigh an individual's energy demands. Such over-consumption where high-energy foods are readily available undoubtedly contributes to the growing problem of obesity. Palatable food stimulates brain circuits similar to those that mediate behavioral responses to drugs of abuse, which may underlie the continuation of food intake long after energy requirements are met. Among the brain areas implicated in reward and food intake, the lateral hypothalamus (LH) has long been recognized as a common region involved in both. It has been suggested that orexin neurons that are expressed exclusively within and adjacent to the LH comprise a major cellular substrate for the functioning of the LH. Here, we review the idea that the orexin neuropeptides play a key role in the rewarding aspects of food intake through interactions with both peripheral and central signals reflecting current energy stores as well as the classic reward pathway--the mesolimbic dopamine system. Furthermore, a possible heterogeneity of orexin neurons is discussed. Uncovering orexin's role in food reinforcement may provide insight into hyperphagia and obesity. In addition, the idea that food intake and substance abuse involve similar brain circuitry suggests potential for a single treatment aiding both obesity and addiction. PMID- 19544626 TI - [The establishment and running status of Chinese measles laboratory anetwork]. PMID- 19544625 TI - Estrogen actions on brain and behavior: recent insights and future challenges. AB - Recent advances in our knowledge of estrogen action in the brain suggest that this steroid is not solely an endocrine factor but plays important but hitherto largely unrecognized physiological and pathophysiological roles that are not directly involved in reproductive processes. Estrogens are now known to influence a wide variety of functions in the mammalian brain ranging from regulation of various aspects of neurotransmitter function and modulation of behaviour to the stimulation of differentiation and plasticity of distinct neuronal populations and circuits. Acting via both genomic and nongenomic mechanisms estrogens can influence higher cognitive functions, pain mechanisms, fine motor skills, mood, susceptibility to. seizures, and also appear to have important neuroprotective function in relation to stroke damage and neurodegenerative disorders. This review focuses on new advancements from clinical and basic studies on estrogen action in the central nervous system especially illustrating the brain regions and cell types in which estrogens produce their effects, emphasizing new knowledge regarding estrogen actions outside the hypothalamus and pituitary gland. Current therapeutic strategies to develop suitable estrogen receptor modulators with a selective spectrum of action in the brain and priorities for future research are also briefly discussed. PMID- 19544627 TI - [Application of a simple method for the detection of measles virus genome by loop mediated isothermal amplification (LAMP)]. AB - OBJECTIVE: A new simple RT-LAMP method was applied to detect measles virus nucleic acid and compared with nest-RT-PCR. METHODS: Compare the detection rate of the RT-LAMP method with that of nest-RT-PCR by detecting measles virus nucleic acid from measles virus and clinical samples. RESULTS: The nucleic acid positive rates of all 23 strains of measles virus are all 100% by the two methods. But to the detection of 18 clinical samples which are negative in measles isolation, the nest-RT-LAMP showed 56.52% positive rate of nucleic acid of measles virus and nest-RT-PCR showed 47.83%. CONCLUSION: RT-LAMP is more sensitive than nest-RT PCR. PMID- 19544628 TI - [Measles pathogenic surveillance from 2005 to 2007 in Guangdong Province]. AB - OBJECTIVE: To develop pathogenic surveillance on measles and to effectively isolate measles virus. To know the genetic characterizations and molecular epidemiology of wildtype measles viruses from 2005 to 2007 in Guangdong Province, and provide the scientific basis for measles control and eradication. METHODS: Vero/Slam cell line were used, measles viruses were isolated from throat swabs or urine specimens collected from uspected measles patients in outbreaks and sporadic patients. A 450 nucleotides fragment of the C-terminal of the nucleoprotein (N) gene was amplified and by RT-PCR and subjected to sequence and phylogenetic analysis using Bio-Edit software. RESULTS: 82 wild-type measels virus were obtained from 377 throat swabs and urine specimens from 2005 to 2007 in Guangdong Province measles lab. The measles isolation rate was 23.58% in 2005, 17.11% in 2006, 39.13% in 2007. The succeed rate of virus isolation is related to the quality of specimens collected and the days after rash occurrence. CONCLUSIONS: We have grasped the technicalability of measles virus isolation and confirm action, and got higher isolation ratio. The wild-type measles virus isolated from Guangdong Province is of H1 genotype from 2005 to 2007, which is the same as the dominant genotype circulation. PMID- 19544629 TI - [Genotype and gene charactrization of wild-type measles viruses circulated in Jilin Province]. AB - OBJECTIVE: To study the genotype and gene characterization of measles wild viruses circulated in Jilin provinces, and to provide scientific evidences for setting down controlling and preventing strategy and measures. METHODS: 38 strains of measles virus isolated in 2001-2006 were genotyped by RT-PCR-RFLP, some strains of measles virus in Jilin province were chosen for the phylogenetic analysis and for the homology analysis of nucleotide and amino acid sequences. RESULTS: All the 38 strains of measles virus were identified as H1 genotype by RT PCR-RFLP, and 29 strains of them were identified further as H1 a by sequence analysis. The homology of nucleotide was 88.0%-89.4% and the homology of amino acid was 91.8%-92.7% .The average diversity was less than 1.4%. CONCLUSION: The measles virus of H1a genotype was the circulating virus within recent years in Jilin province. There were the same measles virus strains circulating and transmitting at different years and also the different H1a measles virus strains co-circulating at the same year. There were the same transmission chain caused by the same measles virus with other provinces. PMID- 19544630 TI - [The epidemiology character of the 503 residual paralysis of acute flaccid paralysis cases]. AB - OBJECTIVE: Analyze the epidemiology character of the residual paralysis(RP) of acute flaccid paralysis (AFP) in Guangdong during 1994-2007. METHODS: The viruses isolated from the excrement of RP cases were identified and typed in Guangdong from 1994 to 2007. Statistics analysis was performed to reveal the relationship among the immunization history,age,gender and the distribution of the etiology. RESULTS: A total of 503 RP cases were reported. 150 of which were isolated with PV and 59 were isolated with NPEV. From 1994 to 2007, The PV isolating rate ranged from 18.92% to 47.06% and was higher in winter and spring, while the NPEV isolating rate ranged from 4.17% to 25.00%. and was higher in summer and autumn. The PV isolating rate decreased as the age increased,and its isolating rate (61.11%) was highest in "0" year group. The PV isolating rate of the population of < or =2 times OPV was far higher than 3 times. The PV and NPEV isolating rate of the RP cases was higher than without RP. CONCLUSION: The case with RP caused by wild poliovirus wasn't found from 1994 to 2007 in Guangdong, but the relationship of RP case was observed between < or =2 years group and < or =1 time OPV, and NPEVs probably are the potential etiological agent that cause children RP. PMID- 19544631 TI - [Immune response induced by recombinant adenovirus combined with recombinant adeno-associated virus type 1 containing HPV16 L1 gene]. AB - OBJECTIVE: To evaluate the immune potency of recombinant adenovirus combined with rAAV1 vector expressing HPV16L1 protein in mice. METHODS: The rAdV and rAAV1 vector containing codon-modified HPV16L1 gene was constructed using Admax and AAVmax packaging system respectively. C57 BL/6 mice were immunized with purified rAdV and rAAV1 vector through intramuscular and intranasal inoculation routes, and the titer of neutralizing antibody was determined by neutralization assay based HPV16 pseudovirus. RESULTS: Intramuscular immunization by rAAV1-mod. HPV16L1 or combined with rAd-mod. HPV16L1 can induce higher titer of neutralizing antibody in serum than that of other groups. The titer of neutralizing antibody of intranasal groups is significantly lower than that of intramuscular group, although the prime-boost strategy using in intranasal group was effective to enhance the specific humoral immunity. CONCLUSION: The rAAV1-mod. HPV16L1 combined with rAd-mod. HPV16L1 can induce higher titer of neutralizing antibody in serum through intramuscular route than that of other groups at the 16th week after the first immunization. PMID- 19544632 TI - [The construction and identification of the PRNP vectors with ubiquitin or the lysosome-targeting signal]. AB - OBJECTIVE: To evaluate PrP expression characteristic of PRNP nucleic acid vaccine vector with ubiquitin or the lysosome-targeting signal. METHODS: The gene of ubiquitin and lysosome-targeting signal were ligated to PRNP and pcDNA3.1 vector that is, pcDNA3.1-UPrP and pcDNA3.1-PrPL were constructed. The expression characteristics of PrP with two signals were evaluated by Western Blot and the localization was observed by indirect immune fluorescence. RESULTS: The protein expressed by pcDNA3.1-UPrP and pcDNA3.1-PrPL with ubiquitin and lysosome targeting signal can be recognized by prion-specific antibody. The protein has three glycosylation molecules form as native PrP.PrP with ubiquitin was degraded gradually with time extension,whereas quantity of PrP with lysosome signal reduced in 48 h after transfection. The protein with two location signals can direct fusion proteins to cytoplasm. CONCLUSION: The PRNP vectors with ubiquitin or the lysosome-targeting signal were constructed and expressed in eukaryocyte successfully. There will be one of good foundation on PRNP nucleic acid vaccine. PMID- 19544634 TI - [Hepatocyte apoptosis in the patients with chronic hepatitis B]. AB - OBJECTIVE: To investigate the relationship between hepatocyte apoptosis and the level of inducible nitric oxide synthase (iNOS) in hepatic tissue in the patients with chronic hepatitis B(Ct-IB). METHODS: We observed 37 cases with CHB and 10 normal controls. Transferase-mediated-UTP-biotin nick-end labling (TUNEL) technique was used to detect apoptosis cells and immunohistochemical staining were also performed to investigate the expression of inducible nitric oxide synthase (iNOS) in biopsy samples. The serum level of ALT, HBV DNA, grading of necroinflammatory activity and staging of fibrosis were also assessed. RESULTS: Hepatocytes in all CHB liver tissues were positively stained by TUNEL in various degree. In contrast, control tissues did not show DNA fragmentation. A significant correlation was seen between apoptosis index (AI) and necroinflammatory grading (r = 0.404, P = 0.015) and serum iNOS level (r = 0.465, P = 0.004). It did not correlate with fibrosis stage and serum alanine aminotransferase level. CONCLUSION: The oxidative stress in patients with CHB may reflected the apoptosis of hepatocyte. Apoptosis involves in liver injury of CHB, but with no significant correlation to serum level of ALT. PMID- 19544633 TI - [Relationships between liver function test, serum HBeAg or HBV DNA level and liver pathological changes in patients with chronic hepatitis B]. AB - OBJECTIVE: The aim of this study was to investigate the relationship between liver function test, serum HBeAg, HBV DNA level and liver pathological changes in patients with chronic hepatitis B. METHODS: 233 patients with chronic hepatitis B accept liver puncture biopsy, liver function test, HBeAg detection and HBV DNA fluorescent quantitation PCR detection. Comparisons of liver function test, HBeAg and HBV DNA level were conducted among different liver pathological changes including inflammation grading and fibrosis staging. RESULTS: In different inflammation grading groups, ALT was highest in group G3 and lowest in group G(0 1)(P = 0.016); TBil was highest in group G4 and lowest in group G(0-1) (P = 0.000); HBV DNA level was highest in group G4 and lowest in group G(0-1), but not statistically significant among groups (P = 0.463). In different fibrosis staging groups, ALT was highest in group S3 and lowest in group S(0-1), but not statistically significant among groups (P = 0.562); TBil was highest in group S4 and lowest in group S2 (P = 0.039); HBV DNA level was highest in group S3 and lowest in group S(0-1), but not statistically significant among groups (P = 0.395). In HBeAg positive group,the proportion of G(3-4) in inflammation grading or S(3-4) in fibrosis staging was lower than that in HBeAg negative group (46% vs. 52%, P = 0.438; 38% vs. 53%, P = 0.025; respectively). CONCLUSION: HBV DNA level can not indicate the severity of liver inflammation or fibrosis in chronic HBV infection. Patients with HBeAg negative often are complicated with more severity of liver fibrosis. In routine liver function test, TBil level correlates with liver inflammation grading or fibrosis staging; ALT level also correlates with liver inflammation grading but not with fibrosis staging. PMID- 19544635 TI - [Construction and identification of replication deficient recombinant adenovirus encoding F gene of subgroup A human respiratory syncytial virus]. AB - OBJECTIVE: A strain of replication deficient recombinant adenvirus encoding fusion glycoprotein (F) of subgroup A human respiratory syncytial virus (RSV) was constructed and the expression of F was identified. METHODS: The F gene was obtained from pGEM3zf-F with Xho I and Hind III, cloned into adenoviruse shuttle vector pShuttle-CMV,and then the resulting pShuttle-CMV/F was transformed into E. coli BJ5183/p with pAdeasy-1 to produce pre-adenoviral plasmid encoding F by homologous recombination. This resultant plasmid was linearized by digestion with Pac I and transfected into 293 packaging cells to generate FGAd-F. Finally, the expression of F protein was identified by Western Blot analysis. RESULTS: FGAd/F was successfully constructed, and the expression of RSV F protein was identified by Western Blot. CONCLUSION: We have obtained a strain of replication-defective adenovirus FGAd/F encoding RSV F protein, which can be used further to investigate its protective efficacy against RSV infection in vivo. PMID- 19544636 TI - [Effect on mRNA and secretion levels of proinflammatory cytokines in DC infected by SARS-CoV N gene recombinant adenovirus]. AB - OBJECTIVE: To investigate the exact mechanism of SARS-CoV pathogenesis at the protein level. METHOD: Under the condition of the establishment of dendrtic cells (DC) culture method, we used recombinant adenovirus to infect mature DC to make clear the development changes in mRNA levels and secreted protein levels of proinflammatory cytokines of IL-1beta, IL-6, IL-8 and TNF-alpha by using RT-PCR and ELISA. RESULT: We found that mRNA levels and secreted protein levels of IL-6 and TNF-alpha in DC had increased gradually after rAd-N infection during first 24 h compared with the control DC infected by rAd-LacZ (P < 0.05 or P < 0.01). CONCLUSION: It is suggested that N protein may be related to the excessive secretion of proinflammatory cytokines during SARS-CoV infection at the acute phase. PMID- 19544637 TI - [Molecular characterization and sequence comparison of the M and S segments of Hantavirus ZJ5 strain]. AB - OBJECTIVE: In order to understand the molecular characters of Hantavirus ZJ5 strain, its complete M and S genome were sequenced and compared with that of other hantavirus strains. METHODS: We prepared the total RNA from ZJ5. Infected cells and the raw or purified RT-PCR product was cloned and sequenced. RESULTS: With sequence compation, we found ZJ5 strain complete M and S segment had higher homology with SEO-type strains than other type of HV, but differential genes were 11.7%-19.2% and 6.7%-14.5% from SEOV. The phylogenetic trees constructed by complete M ind S segment showed that ZJ5 strain was located in SEOV group, and structured alone embranchment. CONCLUSION: The ZJ5 strain was believed to belong to SEO-type virus,and suggest that ZJ5 strain is a new subtype S SEOV group,and structured alone embranchment. CONCLUSION: The ZJ5 strain was believed to belong to SEO-type virus, and suggest that ZJ5 strain is a new subtype from other SEO viruses. PMID- 19544638 TI - [Improved expression of human rotavirus G1VP7 and G3VP7 antigens in the recombinant adenoviruses by codon optimization]. AB - OBJECTIVE: To increase the recombinant adenovirus vector mediated human rotavirus G1VP7 and G3VP7 gene expression through coden optimization. METHODS: We have artificially synthesized two genes of group A human rotavirus that encode G1VP7 and G3VP7 according to the human biased codon. The modified genes were transfected into 293 cells using adenovirus vectors and the gene products, the respective proteins were produced. The expression level of optimized genes and wild type genes was detected by Western Blot. RESULTS: A remarkable increase of the expression level of optimized G1VP7 and G3VP7 genes in comparison with the wild type control. CONCLUSION: The coden optimization indeed help increasing the recombinant adenovirus mediated human rotavirus gene expression, which indicated the potential applicationof such recombinant adenoviruses in the development of adenoviral-vectored rotavirus vaccines. PMID- 19544639 TI - [Occult hepatitis B virus infection in chronic viral hepatitis patients with non A to E hepatitis virus infection]. AB - OBJECTIVE: To observe the status of occult hepatitis B virus infection in chronic viral hepatitis patients with non-A to E hepatitis virus infection and explore the diagnostic value of fluorescence quantitative polymerase chain reaction (FQ PCR) technique for occult hepatitis B virus infection. METHODS: The amount of HBV DNA in serum and liver tissue from 57 patients with non-A to E hepatitis virus infection who were diagnosed as chronic viral hepatitis by Menghini method liver biopsy were detected by using FQ-PCR technique, then the relation between the viral load of HBV DNA in liver tissue and hepatic inflammatory activity were analyzed. RESULTS: Thirteen (22.81%), 22 (38.60%) patients were positive for HBV DNA in serum and liver tissue, respectively. The positive rate and the level of HBV DNA quantity in liver tissue were significantly higher than those in serum; HBV DNA was found positive in both serum and liver tissue in 13 cases, negative in both serum and liver tissue in 35, positive in liver tissue but negative in serum in 9, and in none of the cases HBV DNA was positive in serum but negative in liver tissue (P < 0.01). The logarithmic value of HBV DNA from 13 patients in liver tissue and in serum was respectively: (6.62 +/- 1.21) copies/g vs.(4.03 +/- 1.06) copies/ml, P < 0.01. The hepatic lesions of all HBV DNA positive patients were active pathologic changes, but the level of HBV DNA in liver tissue was not significantly correlated with the grade of hepatic inflammation activity (P > 0.05). CONCLUSION: Occult HBV infection is the etiology of part of the chronic viral hepatitis patients with non-A-E hepatitis virus infection. Missed diagnosis will occur if diagnosis of hepatitis B is only based on detection of serum HBV markers. It is useful for improvement of the diagnostic level of HBV infection via detection of HBV DNA quantitatively in serum especially in liver tissue of chronic viral hepatitis patients with non-A-E hepatitis virus infection by using FQ-PCR technique. The chronic viral hepatitis patients with occult HBV infection should be also given effective anti-viral therapy. PMID- 19544640 TI - [Identification and analysis of an SEO strain of hemorrhagic fever with renal syndrome hantavirus from Guangxi]. AB - OBJECTIVE: To identify hantavirus from lung specimens of rats captured in Guangxi. METHODS: Rats were collected from various areas in Guangxi in combination with plague surveillance and rat lung specimens were examined by ELISA for hantavirus antigen, and M segment of positive specimen was partially amplified with RT-PCR and sequenced. Phylogenetic analysis was conducted for genotyping. RESULTS: From a total of 306 rat lung specimens, a strain of SEO hantavirus was detected from lung specimen of a Norvegicus that was from coastal area of Guangxi Qinzhou city. CONCLUSIONS: For the first time SEO hantanvirus was detected in the coastal area of Guangxi Qinzhou city, its epidemiological significance needs further study. PMID- 19544641 TI - [Immune effects of mutated hepatitis B virus precore-core DNA vaccines in mice]. AB - OBJECTIVE: To observe the immune effect of DNA vaccines encoding mutated HBV pre c/c gene (VE2,VE4) in mice. METHODS: Three kinds of plasmid VEC(DNA vaccines encoding HBV pre-c/c gene), VE2 and VE4 were injected into the thigh muscles of different group of BALB/c mice.Blood and splenocytes from mice were isolated at 4 weeks after immunization. We also have mouse groups immunized with three of these plasmid combined with IFN-gamma gene plasmids. The anti-HBc and anti-HBe antibody in peripheral blood in mice were detected by enzyme linked immunosorbent assay (ELISA), antigen-specific cell immune responses were detected by CTL test and enzyme linked immunospot assay(ELISpot). RESULTS: We found that anti-HBe titers of VE2 and VE4 immunizing groups are higher than VEC group (P < 0.05). We also observed that VE2 and VE4 could induce stronger antigen-specific immune responses than VEC and when combined with IFN-gamma plasmid,the antigen-specific immune responses are stronger than those without combination immunization in mice (P < 0.05). CONCLUSIONS: The DNA vaccine VE2 and VE4 could induces stronger antigen specific immune responses than VEC, and when combined with IFN-gamma plasmid,the antigen-specific immune responses are improved in mice. PMID- 19544642 TI - [Hepatitis viruses infection situation in Mianyang of the Sichuan province]. AB - OBJECTIVE: To investigate the seroprevalence of hepatitis viruses in Mianyang of the Sichuan province. METHODS: EIISA was used for detecting anti-HAV IgG, HBsAg/HBsAb, anti-HCV IgG and anti-HEV IgG of the serum samples. All sample were collected in Mianyang areas in 2007. RESULTS: 1352 samples were detected. The positive rates of anti-HAV, HBsAg/HBsAb, anti-HCV,and anti-HEV are 81.07% (1096/1352), 5.40% (73/1352) and 61.32% (829/1352), 0.37% (5/1352) and 49.26% (666/1352), respectively. The positive rate at different age group, for anti-HAV was 38.21% of 10-19 years old, 83% of 20-29 years old, 88% of 30-39 years old, 95.03% of 40-49 years old, 97% of 50-59 years old, 97.77% of 60-69 years old, 97.52% of > or =70 years old. For HBsAg/HBsAb were 5.65% or 50.83%, 10.0% or 68.0%, 5.20% or 78.80%, 5.97% or 78.11%, 6.50% or 62.50%, 1.12% or 51.40%, 4.96% or 30.58% at the same age group, respectively,for anti-HCV, was 0.33% of 10-19 years old, 0.80% of 30-39 years, 0.56% of 60-69 years old, 0.83% of > or =70 years old.For HEV-IgG was 26.58% of 10-19 years old, 42.0% of 20-29 years old, 55.22%-61.0% of 30-> or =70 years old, for anti-HEV IgM, was 10.06% (53/527) in the positive samples of HEV-IgG. CONCLUSION: The inoculation againt HAV and HBV is enhanced in the young population. HBsAg carrier and HCV infection is decreasing. The HEV infection is actually increasing. PMID- 19544643 TI - [Vasoactive intestinal peptide expression and its clinical significance in gastric adenocarcinoma]. AB - OBJECTIVE: To investigate the expression of vasoactive intestinal peptide (VIP) in gastric adenocarcinoma, and to evaluate the correlation of VIP level with clinical pathologic parameters. METHODS: The level of VIP in sera from gastric adenocarcinoma patients and healthy people was investigated by ELISA. Moreover, the differential gene expression between gastric adenocarcinoma, gastric dysplasia, and the corresponding normal gastric mucosa were determined by RT-PCR. Western Blot was also used to measure the expression of VIP in the gastric adenocarcinoma and the normal gastric mucosa. RESULTS: The serum level of VIP was (5.794 +/- 0.014) ng/ ml in normal control and was (14.437 +/- 0.825) ng/ml in gastric adenocarcinoma patients, showing significant difference (P < 0.05). Meanwhile,the V/B of gastric adenocarcinoma tissues was greater than that of gastric dysplasia and the corresponding normal gastric mucosa (P <0.01), the values of V/B were 1.5261 +/- 0.3028, 0.9334 +/- 0.2872,and 0.9051 +/- 0.2794, respectively. The values of V/B between normal gastric mucosa and gastric dysplasia were not different significantly (P > 0.05). There were significantly negative correlation between the VIP mRNA expression of the differentiation degree of tumor (P < 0.05). The VIP mRNA expression was higher in gastric adenocarcinoma with lymph node metastasis than that without lymph node matastsis (P < 0.05). The VIP protein expression of the gastric adenocarcinoma tissues was greater than that of normal control. CONCLUSION: This findings provide a direct evidence to support the possibility that VIP play a cofactor role in the pathogenesis of gastric adenocarcinoma. PMID- 19544644 TI - [The expression of apoptosis related genes Bcl-2 and Bax protein in dysfunctional uterine bleeding]. AB - OBJECTIVE: To research the expression of apoptosis related genes Bcl-2 and Bax protein in dysfunctional uterine bleeding. METHODS: Detecting the expression of Bcl-2 and Bax protein in 40 cases of endometrium tissue of dysfunctional uterine bleeding patients with immunohistochemistry antibiotics protein-peroxide emzyme (SP)methods. RESULTS: (1) The expression of Bcl-2 protein changes clearly and periodically in endometrium in normal cycle, the difference is obvious (P < 0.05). (2) The Bcl-2 protein develops with the hyperplasia in endometrium and the expression intensity is enhanced, the difference is obvious( P < 0.05). (3) The expression of Bax gene is masculine in endometrium in normal menstrual cycle. (4) The expression of Bax gene descends gradually with hyperplasia in uterine endometrium (P < 0.05). CONCLUSION: Apoptosis of endometrium cell is restrained from the excessive expression of Bcl-2 protein and deficient expression of Bax protein, so the uterine endometrium gets to hyperplasia or notypical hyperplasia. PMID- 19544645 TI - [Protection effects of acetylcholinesterase inhibitors on apoptosis]. AB - OBJECTIVE: To investigate whether tacrine and donepezil can prevent apoptosis induced by Lipopolysaccharides. METHODS: Phase-contrast microscopes was used to observe the morphological changes of Vero cells. Cell counting kit-8 was used to measure cell survival vitro. DNA fragment was analyzed by agarose gel electrophoresis to determine the apoptotic biochemical changes. RESULTS: Vero cells treated with 400 microg/ml and 500 microg/ml Lipopolysaccharides exhibited cell apoptosis. 10 micromol/L tacrine provided protective effect to 500 microg/ml Lipopolysaccharides induced cell apoptosis measured by Phase-contrast microscopes, cell counting kit-8 and DNA fragment analyze. However, donepezil did not show any protective effect of the apoptosis induced by 500 microg/ml Lipopolysaccharides. CONCLUSION: Lipopolysaccharides can induce apoptosis in Vero cells to built an apoptotic model in vitro. Tacrine rather than donepezil can inhibit Lipopolysaccharides induced apoptosis in Vero cells. PMID- 19544646 TI - [Pathogenic diagnosis of an unexplained epidemic event of infectious diarrhea]. AB - OBJECTIVE: To determine the pathogen of an unexplained epidemic event of infectious diarrhea by laboratory diagnosis of suspected cases samples. METHODS: 28 samples from 28 suspected cases (22 fecal samples, 3 vomitus samples, 3 anus swab samples) were tested for Norovirus by RT-PCR. Sequencing and phylogenetic analysis were acomplished of 5 positive samples. RESULTS: 160 of 5694 population were ill with an attack rate of 2.81%. The peak period was 7-9, March. 14 of 28 samples were tested Norovirus positive.Sequencing and phylogenetic analysis showed Norovirus type GII/4 was the causative agent and it had highest identity (97. 9%) with epidemic strain 2006b. CONCLUSION: The epidemic event ofinfectious diarrhea were caused by GII/4 Norovirus strains. PMID- 19544647 TI - [Detection of C-terminal 30 bp-deletion mutation of latent membrane protein 1 of EBV in childhood lymphoma]. AB - OBJECTIVE: To study C-terminal 30 bp-deletion mutation of latent membrane protein 1 of the virus from childhood lymphoma. METHODS: Nested-PCR was used to amplify C terminal of EBV-LMP1 from childhood lymphoma and non-lymphoma associated to EBV, including Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL)and reactive hyperplasia of lymph node (RL). Sequence analysis was performed on the positive PCR product. RESULTS: LMP1 with 30 bp deletion in C-terminal was detected in 11/25 HL,3/8 NHL and 5/15 RL cases respectively. There were no significant differences among HL, NHL and RL (P = 0.793). Sequence analysis showed that LMP1 detected in this study belongs to the following three subgroups: B95.8, China1 and China2. CONCLUSION: LMP1 with 30 bp deletion in C-terminal widely existed in childhood HL,NHL and RL. There was no correlation between special types of LMP1 and the diseases. There were 3 LMP1 subgroups of EBV in children's lymphoma in the cases studied, including B95.8, China 1 and China 2. PMID- 19544648 TI - [Antibody detection and sequencing analysis of hepatitis E virus in human population, swine and chicken in Sichuan region in China]. AB - OBJECTIVE: To investigate the seroprevalence of HEV infection and genotype. METHODS: ELISA were used for detecting anti-HEV IgG of the serum samples, the nested reverse transcriptase PCR (RT-nPCR) was used for detecting HEV RNA in patient serum and swine bile samples. All samples were collected in 2005-2007 in some districts in Sichuan province. The primers used for genotyping were the ORF2 region of HEV genome. RESULTS: The anti-HEV IgG was detected positive in childrens 6.10% (41/672), adults 42.26% (280/ 661), swines 88.89% (32/36), chickens negative (0/59). 1 case of 15 serum samples of anti-HEV IgM positive and 3 of 54 swine bile samples were positive for HEV RNA by RT-PCR.Sequence analysis of 4 isolates has 92.1% to 98.6% nucleotide sequence homology. These isolates from human and swine were identified closely related to Ch-T21 strain 90.1%-96.9% sequence homology, which belonged to HEV genotype 4B. CONCLUSIONS: The swine were the risk factors in the spread of hepatitis E virus. PMID- 19544649 TI - [Relationship between EB virus, Cytomegalo virus, herpes simplex virus and coxsackievirus infection and relapse of multiple sclerosis]. AB - OBJECTIVE: To study the relation between the recent active infection with Epstein Barr virus, cytomegalovirus,herpes simplex virus-1, coxsackievirus B I-IV and the relapse of relapsing-remitting multiple sclerosis (RR MS). METHODS: Using ELISA method, IgM antibodies to Epstein-Barr virus, cytomegalovirus, herpes simplex virus-1, coxsackievirus BI-IV in the plasma from 34 RR MS patients and 200 normal controls were detected. The rates of recent active infection with the above mentioned viruses of the patients and controls were compared.For patients group,comparison was also made between the clinical data of recent active infected patients and patients without recent active infection. RESULTS: There was no statistically significant difference in positive rates of positive rates of IgM antibodies against Epstein-Barr virus, cytomegalovirus, herpes simplex virus-1 and coxsackievirus BI, II, III or VI between the two groups. While there was statistically significant difference in positive rates of IgM antibodies to coxsackievirus B VI and V in the RR MS patients and those in the controls (being 3/34 and 0/200 P < 0.05; 2/34 and 0/200 P < 0.05, respectively). In the patient group, when patients who had active infection with any of the viruses were compared with those who had no active infection, no significant difference between them was found in terms of age, course, frequency, body temperature on admission, differential leukocyte count (neutrophilic granulocyte, lymphocyte and monocytes), use of glucocorticoids, and EDSS point value. CONCLUSIONS: There is a high rate of recent active infection with coxsackievirus B VI and V in RR MS patients at relapsing stage. While the recent virus active infection is unrelated to the severity of the symptoms. PMID- 19544650 TI - [Study of the clinical and laboratory features of hand-foot-mouth disease]. AB - OBJECTIVE: To study the clinical and laboratory features of the mild and severe hand-foot-mouth diseases (HFMD) in Shenzhen in 2008. METHODS: 145 cases were observed in East-Lake Hospital and Shenzhen Children's Hospital. Of the 145 cases, 124 mild cases and 21 severe cases were involved.All the clinical data and laboratory findings were collected and summarized. After collection of the acute and convalescent consecutive stools and peripheral bloods from the patients with HFMDI, EV71 genes were amplified from these samples by RT-PCR. Enterovirus 71 were cultured and isolated using Vero cell line and R&D cell line. RESULTS: The WBC counts and blood glucose levels of the severe cases were significantly elevated, but the ages of the severe ones significantly decreased compared with those of the mild cases (P < 0.05). EV71 genes could be detected by RT-PCR with 35% positive rate in mild cases and 67% in severe cases. The EV71 gene detection rate of the severe cases was significantly increased in contrast to that of the mild ones. The EV71 were isolated and cultured from the stools of 9 patients, one specimens from the dead's stool. Two severe cases died of neurogenic pulmonary edema and brain-stem encephalitis. CONCLUSIONS: EV71 mainly contributes to HFMD and is responsible for death of some severe cases. High fever, less rash, elevated white blood cell counts and blood glucose concentrations as well as age less than 4 years old should be used for prediction of severe cases. PMID- 19544651 TI - [Clinic value of combination of high-risk human papillomavirus test and cervical cytology test in diagnosing cervical lesions]. AB - OBJECTIVE: To investigate the clinic value of combination of high-risk human papillomavirus test and cervical cytology test in diagnosis of cervical lesions. METHODS: Patients underwent physical examination at our department were checked by high-risk human papillomavirus test, cervical cytology test and colposcope from October 2004 to December 2006. Abnormal patients with cervical abnormalities were asked for pathological test.The diagnostic value of cervical lesions among these different methods were compared. RESULTS: Based on the criteria of histopathology, the sensitivity, specificity, positive-predictive value and negative-predictive value of high-risk human papillomavirus test for detecting all cases of CIN II and CIN III were 94.83%, 31.06%, 55.22% and 87.02% respectively, and those of the cervical cytology were 92.10%, 31.06%, 54.50% and 81.43% respectively.Those values changed to 99.65%, 18.55%, 61.46% and 97.62% respectively if two methods were combined. CONCLUSIONS: Human papillomavirus test and cervical cytology test combined with pathological test can improve the detective rate of cervical lesions and facilitate the treatment. PMID- 19544652 TI - [The clinic features and the short-term efficacy of entecavir treatment of the HBeAG negative acute-on-chronic hepatitis B liver failure]. AB - OBJECTIVE: To investigate the clinical features and the short-term efficacy of ETV treatment of 132 HBeAg negative and 51 HBeAg the HBeAg negative acute-on chronic hepatitis B liver failure. METHODS: positive patients with acute-on chronic liver failure were rolled. 84 patients were treated with ETV (0.5 mg daily) and routine supportive therapy. 99 patients in the control group received supportive therapy only. The clinical features and the short-term (180 days) efficacy of ETV were compared between the HBeAg negative and HBeAg positive groups. RESULTS: The age of HBeAg negative patients was significantly higher (P = 0.001). The viral load was significantly higher in the HBeAg positive group (P = 0.001). There was no significant difference in the severity and the mortality in control group between the different HBeAg groups. In the antiviral treatment groups, 20/25 HBeAg positive patients survived and 32/59 HBeAg negative patients survived (P = 0.004). The mortality of HBeAg negative patients in control group was higher than that in treatment group (P = 0.032). In HBeAg negative patients with the HBV DNA <5 log copies/ml and HBV DNA >3 log copies/ml, 10/18 patients in treatment group survived and 6/30 patients in control group survived (P = 0.011). CONCLUSIONS: There was no significant difference in the mortality between different HBeAg patients in routine treatment group. Antiviral therapy with ETV significantly improve survival rate in HBeAg negative patients. In patients with the HBeAg negative and HBV DNA <5 log copies/ml and >3 log copies/ml, antiviral therapy with ETV can improve the survival rate. PMID- 19544654 TI - [In vitro inhibitory mechanism of respiratory syncytial virus with solution prescription of zhidanhuayu]. AB - OBJECTIVE: To study the effective part of solution prescription of Zhidanhuayu (ZDHY) against respiratory syncytial virus (RSV) in vitro. METHODS: Observe the pathology of RSV to Hep-2 under the condition of different concentrations and each effective part of ZDHY. RESULTS: The concentration limit causing celluar toxicity of ZDHY is 5.5 mg/ml. The ZDHY failed to block the absorption of RSV to Hep-2 within this concentration, and consequently the cell fell into the full pathological changes. During the concentration of 2.75-5.50 mg/ml, the ZDHY directly destroyed virus array,meanwhile, the infected cells that treated by the medicine kept healthy also. CONCLUSION: ZDHY could not defend the infection of RSV, but is able to destroy the RSV directly and inhibit the RSV inhabiting in the cell. PMID- 19544653 TI - [Clinilal study of medical ozone therapy in chronic hepatitis B of 20 patients]. AB - OBJECTIVE: To study the effect of medical ozone in the treatment of chronic hepatitis B. METHODS: 42 patients with chronic hepatitis B were divided randomly into two groups. 22 patients treated with basic therapy were as a control group. 20 patients treated with basic therapy plus ozone therapy were taken as a treatment group. Index of biochemistry and virology were studied at initial and post-treatment 8 weeks. RESULTS: After the treatment, liver function of the treatment group and the control group had more significant improvement. The treatment group complete effective and partial effective were 10% and 35% difference. The control group complete effective and partial effective were 4.6% and 13.6% (P < 0.05). CONCLUSION: Treatment of medical ozone on patients with chronic hepatitis B is effective. PMID- 19544655 TI - [Effective inactivatian test of inactivated hepatitis A vaccine using integrated cell culture/strand-specific reverse transcriptase-polymerase chain reaction]. AB - OBJECTIVE: To establish an quick, sensitive and specific assay for effective inactivatian test of inactivated hepatitis A vaccine. METHODS: effective inactivatian test of inactivated hepatitis A vaccine were carried out using integrated cell culture/strand-specific RT-PCR (ICC/strand-specific RT-PCR) assay compared with traditional ELISA and nest RT-PCR assay. RESULTS: all the samples were infectious negative detecting by both ICC/ strand-specific RT-PCR and ELISA assay,while some samples appeared false positive detecting by nest RT-PCR. CONCLUSION: ICC/strand-specific RT-PCR assay is a novel, rapid, sensitive and reliable method for effective inactivatian test of inactivated hepatitis A vaccine. Shorting detection period largely, this assay may be used as an alternative method for routine inactivated hepatitis A vaccines test. PMID- 19544656 TI - [Expression of recombinant VP1 protein of enterovirus 71 and development of serological assay for detection of EV71 infection]. AB - OBJECTIVE: To obtain a recombinant purified Enterovirus 71 VPI protein and establishment of an early, rapid and accurate serological ELISA (enzyme-linked immunosorbent assay) for detection of EV71 infection. METHODS: VP1 gene was amplified by PCR and clonel into pET-21b (+) vector, the positive recombinant plasmid were transformed into E. coli BI21(DE3), and was induced with IPTG, the recombinant protein by SDS-PAGE and Western Blot assays. Finally, the recombinant purified VP1 protein was used as a coated antigen for detection of serum anti-IgM and IgG against EV71 by ELISA. RESULTS: The purified VP1 was obtained, and it can be recognized by sera of patients with EV71 infection associated with hand-foot mouth disease. The A values of anti-EV71 IgM and IgG were significantly elevated as compared to healthy objects and HFMD patients without EV71 infection (P < 0.05). The sensitivity and specificity of IgM to EV71 were 73% and 77% compared with the RT-PCR results, respectively;and those of IgG being 82% and 83%, respectively. CONCLUSIONS: The recombinant protein VP1 was produced and purified, and it was proved to have a good antigenicity and could be used to develop a serological diagnosis kit for EV71 infection in the future. PMID- 19544657 TI - [Research on an improved Lowry method to determine content of protein in Sabin IPV]. AB - OBJECTIVE: To establish a method for the content determination of protein in Sabin IPV. METHODS: Using lowry method combined with being precipitated by trichloroacetic acid to determine the content of protein in Sabin IPV. Changing different conditions to optimize the experiment to establish a improved lowry method. And the sample recovery test was also conducted. RESULTS: The method can exclude the interference of free aminoacid, phenols and some other additives. The calibration curve was in good linearity of protein within the range of 2.5 microg/ml-40 Microg/ml, r = 0.9998. Under the best conditions, the mean recovery was 95.32%, the CV in a batch and between batches were both < 10%. CONCLUSION: The method can be used to determine the micro content of protein in vaccines. PMID- 19544659 TI - [Detection of influenza viruses/avian influenza viruses and identification of virulence using a microarray]. AB - OBJECTIVE: To establish the DNA microarray to detect influenza viruses and avian influenza viruses, and identify their virulence. METHODS: Hemagglutinin (HA), neuramidinase (NA) and nucleoprotein(NP) genes were chosen simultaneously as targets for designing a microarray used for detection of viruses and identification virulence. The nucleic acid were amplified by single primer amplication (SPA). And then its specificity,sensitivity and reproducibility were evaluated. RESULTS: The microarray was able to specially detect H1N1, H3N2, B influenza viruses and H5N1, H9N2 avian influenza viruses. Their limits were 8HAU, 16HAU, 32HAU, and 8HAU, 8HAU respectively. The limit for virulence was 32HAU. When samples were analyzed by both RT-PCR and microarray in parallel, the results agreed in 83.9% (47/56). CONCLUSION: The microarray can detect and distinguish five tested viruses, and especially identify virulence. It not only supplies an assistant tool for clinical diagnosis and control of infectious disease, but also is valuable for controlling and preventing outbreak of avian influenza epidemic. PMID- 19544658 TI - [The development and clinical application of papillomavirus genotyping by HPV DNA suspension array the development and clinical application of papillomavirus genotyping by HPV DNA suspension array]. AB - OBJECTIVE: To develop a new platform for genotyping human papillomavirus(HPV) and to investigate its effect in clinical application. METHODS: By combining L1 consensus PCR and multiplex hybridization using a Luminex xMAP system-based suspension array, we developed a rapid high-throughput assay,the HPV DNA suspension array (HPV-SA), capable of simultaneously typing 30 HPVs, including 18 high-risk HPV genotypes and 12 low-risk HPV genotypes. 810 clinical specimens were used to investigate the effect of HPV-SA. Veracity of the genotyping result was verified by E7 type-specific PCR-DNA sequencing. RESULTS: Among the 810 clinical specimens, 243 were found to be HPV positive,including high-risk HPV subtypes 16, 18, 26, 31, 33, 39, 45, 51, 53, 56, 58, 59, 66, 68, 73 and 82,and low-risk HPV6, 11, 34, 54, 61, 67, 70 and 84. The sensitivity tested by standard samples was up to 10 copies of HPV DNA. CONCLUSION: The HPV-SA developed here showed high sensitivity and specificity, suitable to be applied in clinical practice for HPV diagnosis and investigation on the prevalence of HPV sub-types. PMID- 19544660 TI - [Establishment of a fluorescent quantitative PCR detection method for rabies virus and preparation of RNA positive controls]. AB - OBJECTIVE: Establish the fluorescent quantitative RT-PCR detection method for rabies virus (RV) and construct RNase-resistant virus-like particles as positive controls. METHODS: Analyze the database in GenBank, the probe and the primers were designed in the conservative region of N gene of rabies virus and the method of real-time fluorescent quantitative PCR was obtained; On the basis of MS2 phage, with the technology of gene recombination, prepare the RNase-resistant virus-like particles for RV positive controls; RESULTS: RNase-resistant virus like particles were obtained after prokaryotic expression in E. coli. The designed primers and probe were confirmed to be very specific and conservative, and be sensitive to-concentration of 15 copies/microl. CONCLUSION: Established the method of detecting rabies virus by reverse transcription real-time quantitative PCR,obtained the RNase-resistant and no infectivity virus-like particles as positive controls of rabies virus. PMID- 19544661 TI - [Atomic force microscopy investigation of the denudation of the envelope of influenza virus treated with Nonidet P-40]. AB - OBJECTIVE: Through observing the morphology and topography of the prepared influenza viruses (H1N1) treated with the different Nonidet P-40 solutions using atomic force microscopy (AFM), to explore the application of AFM on the research of the internal character of viral morphology and structural virology. METHODS: The virus samples were treated with serial diluted Nonidet P-40 solutions from 0.05% to 0.20% and then investigated by AFM with the tapping mode in air at room temperature to obtain the morphology and topography changes including height data,amplitude data and phase data for both spherical and filamentous influenza virus A. RESULTS: The serial AFM images show that the erosion degree of the virions is proportional with the improvement of NP-40 concentration,and partly denuded virion image appeared at 0.05% NP-40 treatment, which was revealed clearly on both amplitude images and phase images. CONCLUSION: This work demonstrated for the first time that the internal topography of influenza virion could be revealed by AFM via suitable nonionic surfactants chemical dissection. PMID- 19544662 TI - [Detection and identification of human metapneumovirus by real time reverse transcription PCR]. AB - OBJECTIVE: To develop a rapid, sensitive and specific real time reverse transcription PCR for detecting and identifying human metapneumovirus. METHODS: The Hmpv-L gene of human metapneumovirus was chosen as target gene, the primers and TaqMan probe were designed, and the PCR reaction was optimized systematically. The total RNA was extracted from respiratory specimens, and reverse transcription was performed through random primer. The cDNA was detected by using real time PCR. The specificity, sensitivity and reproducibility of real time PCR were estimated. The real time PCR was applied to detect 180 clinical respiratory specimens. RESULTS: The human metapneumovirus can be detected using real time reverse transcription PCR accurately and quickly, and the sensitivity was 1 copy/microl. The coefficient of variation of intra-assay and inter-assay was less than 5%. Among those 180 specimens, 28 (15.56%) were positive for human metapneumovirus, the clinical diagnoses for these 28 patients were pneumonia (15.60%, 17/109) and bronchiolitis (15.49%, 11/71). 21 positive specimens were from patients under 2 years of age, and 6 positive specimens were from patients between 2 and 5 years of age, only 1 positive specimens was from patients over 5 years. CONCLUSION: It is demonstrated that real time reverse transcription PCR is a reliable, accurate and feasible assay for human metapneumovirus, which has become one of the most important pathogens induced acute respiratory infections in pediatric patients. PMID- 19544663 TI - Evolution of pharmacogenomics. AB - Pharmacogenetics dates back more than 2,000 years to observations by Pythagoras, however it was not until the 1950s when some enzyme polymorphisms (e.g., N acetyltransferase, G6PD) were discovered that the term was coined by Vogel. Pharmacogenetics then went into decline as being too esoteric a subject. In the 1970s the discovery of the CYP2D6 polymorphism and its resultant effect on drug toxicity and response led to many observations of pharmacogenetic-based variations in pharmacokinetics. These and other discoveries and the subsequent ability to genotype led to the term pharmacogenomics. Today, there are an increasing number of genes for which polymorphisms have been identified that are associated with variable drug response whether it be at the drug metabolizing enzyme, transporter or receptor level and, mainly through a candidate genes(s) approach. Increasing use of genome-wide analysis is identifying hitherto unpredictable new genes associated with disease and drug response. Although some old and most new drugs coming onto the market have a "pharmacogenomic footprint", the clinical and practical usefulness of pharmacogenomics has been generally lacking. To date, clinical translation of pharmacogenetics has focused on narrow therapeutic index drugs for toxicity (e.g., azathioprine) and more recently for efficacy and toxicity (e.g., warfarin) purposes. Pharmacogenetics and genomics will be advanced through lower cost, rapid whole genome sequencing methods combined with sophisticated algorithms allowing individualised dosage recommendations but not necessarily their adoption. However, complicating this is the influence of changes in gene expression by environmental and genetic factors. Therefore translation of pharmacogenetics into "personalised medicine" will depend on many factors including clinical relevance, environmental-genetic interactions, cost and education. PMID- 19544664 TI - Pheno- or genotype for the CYP2C19 drug metabolism polymorphism: the influence of disease. AB - A genotyping approach for pharmacogenes such as CYP2C19 is often advocated to "personalise therapy", since individuals with the homozygous null allele genotype are poor metabolisers (PM) of many drugs. The genotype-phenotype relationship is a validated approach for S-mephenytoin, proguanil and omeprazole in young, healthy populations. However, this relationship may not be valid in patients with diseases such as cancer and congestive heart failure, or in old age. The high phenotypic discordance in the genotypic EM group means that in many clinical situations the true number of CYP2C19 poor metabolisers may be under-estimated if only genotypic approaches are used. PMID- 19544665 TI - Characteristics of agonist-induced Ca2+ responses in diseased human valvular myofibroblasts. AB - Valvular myofibroblasts (VMFs) are present in large numbers in the cardiac valve. Although the functional roles of VMFs in vivo remain to be determined, evidence suggests the cells' ability to contract in vitro. Since Ca2+ is important in the contractility of many cell types, we examined Ca2+ responses induced by different agonists in normal and rheumatic VMFs. The agonists used were histamine, adenosine triphosphate (ATP) and 5-hydroxytryptamine (5-HT), all of which are important mediators in cardiac function. Cytosolic Ca2+ concentrations ([Ca2+]i) in fura-2-loaded VMFs were measured with ratiometric fluorescence microscopy. VMFs were challenged with a single concentration of each agonist in either Ca2+ containing (+Ca) or Ca2+-free (-Ca) physiological salt solution (PSS). From the resulting Ca2+ response, area under curve (AUC) was calculated from the point of drug addition (i.e., baseline) until the response reached the first peak (i.e., maximum). Our data show that more Ca2+ was mobilized in normal than in rheumatic VMFs, suggesting possible Ca2+-mobilizing dysfunction in the initial phase of a response under disease conditions. The most prominent difference was observed with 5-HT stimulation in +Ca PSS, where normal VMFs showed significantly greater AUC than rheumatic VMFs. The investigation of agonist-induced Ca2+ signaling characteristics in VMFs may provide information pertaining to Ca2+-associated changes and their consequences in cardiac valvular diseases. PMID- 19544666 TI - Effects of inhibitory amino acids on adenosine release in the mouse hippocampus. AB - Adenosine is a neuromodulator which inhibits the synaptic release of neurotransmitters. However, only little is known of the effects of inhibitory neurotransmitters and modulators on adenosine release. We studied these effects with hippocampal slices from developing (7-day-old) and young adult (3-month-old) mice under normoxic and ishemic conditions. In normoxia, adenosine release was about 3-fold greater in adults than in developing mice. beta-Alanine and L-serine (both 0.1 mM) significantly reduced the release of [3H]adenosine in normoxia in developing mice. Glycine, beta-Alanine and L-serine (all 0.1 mM) had similar effects in ischemia. Taurine depressed concentration- dependently the release in developing mice but had no effect in adults. The simultaneous release of taurine and adenosine may constitute an important protective mechanism under ischemic conditions. PMID- 19544667 TI - Gender differences in brain peripheral benzodiazepine receptor (PBR) expression and seizures produced by heptachlor during development. AB - Heptachlor has been widely used as an insecticide. It is a GABA-A antagonist and causes seizures. It also increases peripheral benzodiazepine receptors (PBRs) in brain. PBRs are found on the outer mitochondrial membrane in glia, rather than in neurons, and are necessary for steroidogenesis in brain. We compared the effects of acute oral administration of heptachlor (60 mg/ml oil/kg body wt) at 10 ages from postnatal day (PND) 0 to 60 on brain PBR expression and seizure severity in both male and female rats at 1 and 2 hr after administration. From PND 10 through 60, brain PBR expression was increased about 175-225% of controls at both 1 and 2 hr after heptachlor in females. In males however, PBRs were only increased at 30 60 days at 1 hr but not at any age at 2 hr. At 2 hr after heptachlor at 30-60 days in males, PBRs were significantly lower than at 1 hr and even tended to be lower than control levels. By contrast, seizure intensity was greater in males than in females from 10 through 20 days of age at 1 hr and was even greater at 2 hr from 16 through 30 days of age, reflecting the lower PBR levels at 2 hr than at 1 hr in males. Thus, the gender difference in PBR expression was the opposite of the gender difference in seizure intensity. PBRs in brain synthesize several neurosteroids, including allopregnanolone, which is a potent anticonvulsant agent. We hypothesize that the gender differences in seizure intensity after heptachlor were due to the action of heptachlor in greatly increasing PBR expression in females but not in males. Thus the greater expression of PBRs in females would result in more synthesis of allopregnanolone than in males. Therefore, because of allopregnanolone's anticonvulsant effects, seizure intensity was less in females than in males. By comparison, maximal electroshock (MES) caused seizures and increased PBRs in brain in both male and female developing rats with no gender differences at 10-20 days of age. PMID- 19544668 TI - Agonist-specific regulation of inositol phosphate metabolism in cardiac endothelial cells. AB - The actions of nucleotides and hormones at endothelial cell (EC) receptors are known to result in the release of ATP that acts as a local hormone to facilitate release of mediators such as NO and PGI2. Stimulation of ECs with the P2Y1 receptor agonist 2-MeSATP leads to the rapid release of Ca2+ from stores consistent with a role for inositol trisphosphate (Ins-1,4,5-P3) in mediating the action of extracellular nucleotides. Guinea pig ECs were grown in primary culture. [3H]d-myo-inositol (30 Ci/mmol) labeling studies revealed maximal incorporation of radioactivity into [3H]Ins-1,4,5-P3 when glucose in the labeling buffer was lowered to 1 mM and non-radioactive inositol was added at 10 microM. Stimulation of EC for one sec led to the dose-dependent accumulation of [3H]Ins 1,4,5-P3 as well as [3H]IP4, [3H]IP5, and [3H]IP6. Unexpectedly, the metabolism of [3H]Ins-1,4,5-P3 to IP1 was disparat in stimulated versus un-stimulated cells. In [3H]d-Ins labeled stimulated EC or in homogenates derived from unlabeled, stimulated EC, dephosphorylation of [3H]Ins-1,4,5-P3 led to the exclusive formation of [3H]Ins-4-P1. Addition of on-nucleotide agonists such as bradykinin gave the same results suggesting that the dephosphorylation pathway for IP formation in EC is dependent on agonist stimulation and may be correlated with regulation of agonist responsiveness or heretofore unrecognized actions of IP isomers in stimulated versus unstimulated cells. PMID- 19544669 TI - Levels of glutathione and some biogenic amines in the human brain putamen after traumatic death. AB - Mexico City is among the world's largest metropolitan city centers and one of the most difficult and challenging cities in which to drive a motor vehicle. During peak transit hours and maximum congestion, numerous accidents occur, many of them fatal. The aim of the study presented here was to analyze the levels of select indicators against oxidative stress and levels of biogenic amines as a consequence of accident or altercation and fear deaths. Eighteen cases were studied (sixteen males, two females). Subjects ranged from twelve to eighty-one years of age. Nine of the deaths studied were the result of motor vehicle or subway accidents. Eight of the eighteen deaths were the result of a violent altercation, while one of the deaths resulted from a drug overdose and cardiac arrest. Biopsies of cadaver putamen were homogenized and analyzed for Tryptophan (Trp), 5-hydroxyindole acetic acid (5-HIAA), Dopamine (DA), and Glutathione (GSH) levels by fluorometric methods. Trp, 5-HIAA, DA, and GSH levels showed an increase in the subjects who's death was caused by violent altercation combined with fear, while DA levels showed significant differences in all accident groups. This data suggest that biogenic amines in cadaver putamen tissue, such as DA, can be telling biochemical markers, indicative of altercation and fear deaths. PMID- 19544671 TI - Parenteral nutrition given to children and adults: experience of a procedure applied in Mexico. AB - Nutritional support is a critical step in caring for hospitalized patients both to avoid possible metabolic alterations that would worsen the patient's condition, or as a direct result of a particular disease. The purpose of the present study was to describe a procedure for the prescription of total parenteral nutrition (TPN), its administration, monitoring and the complications experienced in a third level hospital in Mexico, as applied to pediatric and adult patients given TPN. The study was carried out for a period of 30 months. TPN was prescribed according to the clinical status of patients. The study reviewed 4,000 parenteral nutrition records from January 2005 to June 2007 (30 months). Based on data here presented a guideline was applied to improve the nutritional support of patients as part of the need to ensure their recuperation during their hospitalization. We observed that TPN must be individualized, based on daily nutrient recommendations, which can be useful to assess the nutritional status of the hospitalized patient with diverse pathologies. PMID- 19544672 TI - Oral calcium administration attenuates thrombocytopenia in patients with dengue fever. Report of a pilot study. AB - Global climate change is one of the instigating and contributing factors for epidemic outbreaks of infectious diseases in human populations. In the years 2003 to 2005 the city of Tampico, in the northern state of Tamaulipas, Mexico, experienced recurrent outbreaks of dengue virus infections (DV) and the resulting dengue fever (DF). One of the hallmark symptoms of DF, which appears to worsen as the environmental temperature increases, is thrombocytopenia. In as much as it is a hallmark for hemorrhagic manifestations, thrombocytopenia is a useful sign to monitor the course of infected patients. Extracellular calcium (Ca2+ o) plays a key role in blood clotting; its chelation in vitro with ethylenediaminetetracetic acid (EDTA) or citrate prevents clotting, while exogenous recalcification of plasma leads to shortening of clotting time. In vivo, Ca2+ o is essential for platelet function and for the regulation of the immune response. In this work we report a significant increase (p<0.05) in the number of blood platelets of patients with clinical signs and symptoms of DF following oral administration of calcium carbonate (CAL, 1.2 to 1.8 g/day; n=10) when compared with a control group (CTL, n=10): 89 (46-132) versus 206 (155-257). Data expressed as mean value (95% confidence interval, C.I.) for x1000 cells/mm3. CAL also improved overall clinical condition and reduced by 36 % the duration of signs and symptoms of DF: 6.7-11.3 days, versus 11.5-16.6 days (95 % C.I., p<0.05) when compared with CTL patients. The possible mechanism of calcium attenuated thrombocytopenia and clinical improvement is discussed. PMID- 19544670 TI - Inhibition of Nm23H2 gene product (NDPK-B) by angiostatin, polyphenols and nucleoside analogs. AB - Human breast cancer cells (MDA-MB-435s) secrete a nucleoside diphosphate kinase (NDPK-B) as a phosphoprotein capable of converting diphosphate nucleosides to triphosphate nucleotides for one round in the absence of a phosphoryl donor. Incubation of the partially purified NDPK-B (Nm23-H2 by Western blot) from [gamma32P]Pi-labeled cells with non-radioactive ADP results in the formation of [gamma32P]ATP (Proc. West. Pharmacol. Soc. 44: 61-63, 2001). The presence of a secreted protein that can maintain ATP levels in the vicinity of capillary and lymph vessels may support cancer metastasis in several ways based on the known actions of ATP at P2Y receptors: facilitate intravasation of breast cancer cells that migrate from a solid tumor, support their extravasation at a distal site, and stimulate angiogenesis. The putative role of angiostatin (AS) as an ATP synthase inhibitor led us to test the notion that AS blocks NDPK-B activity. Addition of commercial AS (kringles 1-4) did not alter enzyme activity. However, AS produced by us and never lyophilized, blocked NDPK activity in a dose dependent fashion consistent with the notion that extracellular ATP generation by tumor cells may be important to the development of metastases. The ability of 0.5 mg/ml angiostatin to block NDPK-B activity to approximately 75% of control activity compared poorly with the polyphenol inhibitors of. The catechin gallates, theaflavins and ellagic acid inhibited NDPK-B completely with the rank order of potency: EA > theaflavins > EGCG > ECG > PAPS. Our results suggest that the biological activity of angiostatin as a putative metastasis inhibitor may be in part the result of nm23 inhibition and that the production, lyophilization, packaging or storage of commercial angiostatin leads to the alteration of its biological activity against NDPK-B. Ellagic acid is a potent (IC50 = 10.5 microM) NDPK-B inhibitor that may prove useful in elucidating the role of cancer-cell secreted NDPK-B in tumor development. PMID- 19544673 TI - A case of serotonin syndrome associated with methadone overdose. AB - A chronic pain patient prescribed 20 mg of methadone per day was seen at the Emergency Department within one hour following a witnessed intentional 200 mg ingestion. In addition, he was taking the serotonin re-uptake inhibitor antidepressant drugs, sertraline and venlafaxine as prescribed. Methadone is also a serotonin re-uptake inhibitor which has been involved in serotonin toxicity reactions. Initially, no symptoms of narcotic overdose (depressed central nervous system, respiration, or blood pressure) could be distinguished, and the standard narcotic urine screen was negative. No decontamination or antagonist therapy was given, and the patient was discharged to a psychiatric unit for observation. At 5 hours post-ingestion he presented in a panic with hallucinations and elevated blood pressure, pulse, and respiration. These symptoms are characteristic of serotonin syndrome which is often described as mental status changes, autonomic hyperactivity, and neuromuscular abnormalities. At 10 hours post-ingestion the patient was found unconscious. He had aspirated stomach contents into his lungs. His respiration, blood pressure, and pulse were all severely depressed. He never regained conciousness, and he died 5 days later. The medical examiner's finding was probable acute methadone intoxication. In this case serotonin syndrome appears to have opposed and delayed typical narcotic symptoms. Methadone has additional pharmacologic and toxicologic properties which may complicate the assessment and treatment in overdose situations. PMID- 19544674 TI - Coronary artery disease risk factors in patients with diastolic dysfunction. AB - Diastolic dysfunction (DD) is a common entity associated with advanced age and hypertension, often present in patients with ischemic heart disease. The correlation of DD and coronary artery disease (CAD) in asymptomatic young patients is not known. We sought to determine the prevalence of CAD risk factors in asymptomatic patients with DD. We present data from a retrospective chart review of CAD risk factors in asymptomatic patients diagnosed with DD during an echocardiograph examination. We screened all patients that had an echocardiographic examination at Lenox Hill Hospital from January 2004 until July 2007. DD was diagnosed if an impaired filling pattern with an E/A ratio less than one was noted on the mitral inflow pulse wave doppler. One-hundred and one patients met study criteria. Data regarding the presence of the following risk factors was collected: sex, hypertension, hyperlipidemia, smoking, diabetes, peripheral artery disease, and family history of CAD. The mean age of patients was 48 +/- 6; 50% were male. The most prevalent CAD risk factor in this group of patients with DD was hypertension (53%), followed by hyperlipidemia (31%) and diabetes (22%). These data can help identify those patients who are asymptomatic in terms of DD who may be at risk for the condition. These patients should undergo an echocardiograph examination in order to rule out presence of DD, even if symptoms are not present. PMID- 19544675 TI - Takotsubo cardiomyopathy: case report and review of the literature. AB - Takotsubo cardiomyopathy, also known as stress induced cardiomyopathy and transient left ventricular apical ballooning syndrome, is a rare syndrome that is characterized by a transient decrease in ejection fraction. This is accompanied by hypokinesis of the left ventricle and ballooning of the apex, with hypercontractile base and non-obstructive coronary artery disease. Takotsubo was first described in Japan in the early 1990's. It was named such due to the morphology of the apical ballooning that is similar in shape to a 'takotsubo', which is a pot with a round bottom and narrow neck used for trapping octopuses. Though most often described in Asians, reports of Takotsubo in Caucasian populations is becoming more common, possibly due to heightened awareness and detection. The most common presenting symptom of Takotsubo is acute chest pain mimicking myocardial infarction. Patients may also present with dyspnea, pulmonary edema and, more rarely, cardiogenic shock. Clinical symptoms are accompanied by transient left ventricular dysfunction. Despite clinical symptoms consistent with acute myocardial infarction, normal coronary arteries are usually detected upon cardiac catheterization. The case presented here is of an Asian woman who developed symptoms of acute myocardial infarction during a stressful hospital stay. PMID- 19544676 TI - The influence of malnutrition on the oral pharmacokinetics of nimesulide in rats. AB - Malnutrition is a health problem in Mexico. It has been established that malnutrition may produce important changes in the pharmacological response to drugs, since changes in the pharmacodynamics and pharmacokinetics may occur. It has been described that a reduction of plasma proteins and in hepatic enzymes may occur. Due to these changes, absorption, distribution and elimination of drugs can be modified. Nimesulide is a non-steroidal anti-inflammatory agent that is widely used. This drug is importantly bound to plasma proteins and is metabolized through cytochrome P-450, two systems that are altered in malnutrition. In order to establish if malnutrition can modify the pharmacokinetics of nimesulide, a comparison of pharmacokinetic parameters obtained in control and protein-calorie malnourished rats was carried out. Two groups of 7 rats were employed in this study. At 45 days of age, group 1 received a standard balanced diet for 4 weeks, whereas, group 2 received a low protein diet for the same period. Then, rats received an oral dose of 10 mg/kg nimesulide and blood samples were drawn at selected times for 12 hr. Nimesulide whole blood levels were determined by HPLC and the pharmacokinetic parameters; Cmax 1.18 +/- 0.13 and 1.03 +/- 0.10 microg/ml, tmax 5.25 +/- 1.03 and 7.48 +/- 1.09 h and AUC12h 8.64 +/- 1.19 and 8.27 +/- 0.85 microg x h/ml were obtained. We conclude that malnutrition does not modify the oral pharmacokinetics of nimesulide. PMID- 19544677 TI - Splenic non-Hodgkin's lymphoma presenting as recurrent kidney stones -- an "incidentaloma"? AB - Splenic lymphoma, or primary malignant lymphoma of the spleen (PMLS), is an uncommon condition whose true nature is difficult to define due to the variable ways it has been classified. Out of all non-Hodgkin's lymphomas it comprises less than 2% of cases. Some experts suggest that PMLS only involves the spleen and splenic hilum, while others consider PMLS to be an entity that develops within the spleen and later has the potential for invading adjacent organs and metastasizing. Clinical features of splenic lymphoma are characterized by nonspecific systemic symptoms such as low grade fevers, night sweats and symptoms related to considerable splenomegaly. Most of these lymphomas are of B-cell origin showing low or intermediate-grade lymphoma on histological analysis. The case we present here is of a patient presenting with left sided flank pain, and given a previous history of nephrolithiasis, a presumably simple diagnosis of kidney stones was made. However, further investigation led to the discovery of splenic lymphoma, which was asymptomatic earlier but may have manifested symptoms that mimicked renal colic. PMID- 19544678 TI - Comparison of suspension composition on the pharmacokinetics of nimesulide in rats. AB - Nimesulide is a non-steroidal anti-inflammatory agent that is widely used in the treatment of inflammatory pain. The drug belongs to the class II of Biopharmaceutical Classification System (low solubility, high permeability) and therefore, absorption of this drug is limited by its dissolution. It has been established that complex formation of insoluble substances with cyclodextrins may increase their oral bioavailability since solubility is improved. In order to provide test this hypothesis, a comparison on the oral pharmacokinetics of two suspensions of nimesulide in rats was carried out. Two groups of 7 rats were employed. One group received an oral dose of 10 mg/kg of a suspension prepared with 0.5% carboxymethylcellulose solution in water, whereas the other group received a commercially available formulation containing the complex of nimesulde beta cyclodextrins (Eskaflam). Blood samples were obtained at selected times for a period of 12 hours and analyzed by an HPLC method. Pharmacokinetic parameters obtained were as follows: Cmax 1.18 +/- 0.16 and 1.93 +/- 0.12 microg/ml, tmax 5.25 +/- 1.03 and 3.21 +/- 0.91 h and AUC 8.65 +/- 1.19 and 13.74 +/- 0.70 microg hr/ml for carboxy-methylcellulose and Eskaflam, respectively. Values for Cmax and AUC12hr were increased and a reduction of tmax was observed indicating improved absorption of nimesulide in the formulation containing beta-cyclodextrins. PMID- 19544679 TI - Platelet aggregation inhibition in patients receiving statins either fully or partially metabolized by CYP3A4. AB - Clopidogrel therapy is the standard for prevention of cardiovascular thrombotic events. Clopidogrel is converted to an active thiol by the cytochrome P450 CYP 3A4 and 2C19 enzymes. Recent studies suggest that statins metabolized by CYP3A4 attenuate the anti-aggregatory effect of clopidogrel. We evaluated the effect of CYP3A4-metabolized statins (atorvastatin, group 1) and partially-CYP3A4 metabolized statins (simvastatin, group 2) on platelet aggregation inhibition (PAI) when given concomitantly with clopidogrel as compared to patients who were statin naive (group 3). PAI was measured by PlateletWorks (Helena Laboratories ICHOR) using the platelet P2Y12 receptor agonist ADP (20 micromol). All patients were on clopidogrel therapy (75 mg/day). Non-responsiveness was defined as a PAI of < 35%. There was no statistical difference in mean PAI among groups; a higher prevalence of clopidogrel non-responders was noted in group 1 compared to group 3 (p=0.002). Multivariate analysis, adjusting for unequal presence of metabolic syndrome and hypertension, we found no statistical difference between groups. Our data suggests that statins, either fully or partially metabolized by CYP3A4, do not influence PAI when clopidogrel is used at 75 mg/day, even after adjusting for risk factors. We concluded that concomitant statins with clopidogrel therapy does not influence the effect of clopidogrel in PAI. PMID- 19544680 TI - Comparison of the oral pharmacokinetics of fluconazole and itraconazole in Mexicans. AB - Fluconazole and itraconazole are antimycotics widely used in Mexico. However, limited information about their pharmacokinetics is available. It has been reported that physicochemical characteristics of these compounds are disparate, leading to different pharmacokinetic profiles. Moreover, it has been suggested that pharmacokinetics of some drugs may vary in Mexicans when compared with Caucasians due to reduced metabolism by CYP3A4. Based on these distinctions, it is important to carry out local studies in order to establish dosage regimens according the characteristics of each population. The purpose of this study was to compare the oral pharmacokinetics of fluconazole and itraconazole in Mexicans and to compare our results with those reported in other populations. Two groups of 16 subjects volunteered for this study that was approved by the Institutional Research and Ethics Committees. All subjects gave written informed consent for participation. After an overnight fast, volunteers received an oral dose of 100 mg fluconazole or itraconazole and blood samples were obtained at selected times over 96 hr. Plasma was obtained and analyzed by HPLC and pharmacokinetic parameters were obtained. As expected, fluconazole plasma levels were higher than itraconazole due to a lower volume of distribution. Additionally, less variability was observed for fluconazole. When data obtained in Mexicans was compared with those obtained in other populations, no differences were observed, suggesting that there are not interethnic differences in the pharmacokinetics of fluconazole and itraconazole. PMID- 19544681 TI - Herpetic and candidal infections of the esophagus in an elderly male. AB - We present the unique case of an eighty-nine-year-old male without any immunodeficiency state or taking immunosuppressive medication and who did not have conditions affecting clearance of the esophageal lumen who was diagnosed with simultaneous herpetic esophagitis and candidal duodenitis. PMID- 19544682 TI - A pilot study of the effect of diclofenac with B vitamins for the treatment of acute pain following lower-limb fracture and surgery. AB - The aim of this pilot study was to compare the efficacy and tolerability of the non-steroidal anti-inflammatory drug (NSAID), diclofenac (2-(2,6-dichloranilino) phenylacetic acid), for treatment of acute pain originated by lower-limb fracture and surgery, with that of diclofenac plus B vitamins. This clinical trial was single-center, prospective randomized and double-blinded. After giving informed consent, patients with lower-limb closed fractures rated their pain on a 10-cm visual analog scale (VAS). Patients were then randomized to receive 75 mg diclofenac or 75 mg diclofenac plus B vitamins (thiamine, pyridoxine and cyanocobalamin) twice daily (all intramuscularly). Patient evaluations of pain intensity were recorded throughout two periods: twenty-four hours pre-surgically and twenty-four hours postsurgical. Twenty-four hours after the first drug administration, patients underwent elective lower-limb surgery. Standardized general anesthetic techniques were used for all patients. Fourteen patients completed the study. The subjects' assessments of limb pain on the visual analog scale showed a significant reduction from baseline values regardless of the treatment group when surveyed at 12, 24, 36 and 48 hr post operation. All treatments showed a similar profile in pain reduction. There were reports of pain in the administration site, but in general, all the regimens were well tolerated. PMID- 19544683 TI - Persistent pulmonary hypertension in the newborn: therapeutic effect of sildenafil. AB - Persistent pulmonary hypertension of the newborn (PPHN) is a cardiopulmonary disorder characterized by systemic arterial hypoxemia secondary to elevated pulmonary vascular resistance with resultant shunting of pulmonary blood flow to the systemic circulation. PPHN is a serious illness that becomes progressively worse and is sometimes fatal. Management of the disease includes treatment of underlying causes, sedation and analgesia, maintenance of adequate systemic blood pressure, and ventilator and pharmacologic measures to increase pulmonary vasodilatation, decrease pulmonary vascular resistance, increase blood and tissue oxygenation, and normalize blood pH. Inhaled nitric oxide (NO) has been one of the latest measures to successfully treat PPHN. If all other treatments fail, extracorporeal membrane oxygenation (ECMO) can be used. Recently, preclinical and clinical studies have demonstrated the utility of the selective inhibitor of phosphodiesterase type 5 (PDE5), sildenafil, in decreasing pulmonary hypertension (HP). Sildenafil was first employed to offset rebound pulmonary hypertension in infants upon withdrawal of NO treatment. Later, several case reports demonstrated the effectiveness of sildenafil in the treatment of PPHN. Two randomized blinded studies with sildenafil in infants with PPHN were published in 2006. In both studies, the patients treated with sildenafil showed a steady improvement in pulse oxygen saturation over time. Likewise, the frequency death was lower in the groups treated with sildenafil. Recent evidence shows the utility of sildenafil in the treatment of PPHN. However, since existing data are limited there is an urgent need for multicenter blinded, placebo controlled, randomized trials. PMID- 19544684 TI - Nitric oxide stimulation of cGMP accumulation in myometrial cells from pregnant women is antagonized by oxytocin. AB - The role of cGMP in the myometrium of pregnant women is not completely known. We have previously shown in guinea pig, monkey and man that NO-induced relaxation of oxytocin-induced contractions is independent of cGMP accumulation. To approach an understanding of the role of cGMP in myometrium, we have developed smooth muscle cell cultures from pregnant women undergoing caesarian section at term. Cells, grown in standard media containing progesterone, express smooth muscle cell markers and are used within five doublings. Cells stimulated with NO donors increase their cGMP levels nearly 100 fold (basal = approximately 9 pmol/mg protein). In the presence of oxytocin (OT; 1 microM), cGMP accumulation in the presence of NO (100 microM) is significantly blunted (25 fold). Cyclic GMP degradation is inhibited by the presence of the phosphodiesterase inhibitor zaprinast; suggesting that the ability of OT to attenuate cGMP accumulation is unlikely to be due to degradation. We propose that the elevation of intracellular calcium following the addition of OT suppress the activity of a calcium-sensitive guanylyl cyclase. The diminution of cGMP synthetic potential in myometrial cells from pregnant women is consistent with the absence of a role for cGMP in the NO induced relaxation of uterine muscle. PMID- 19544686 TI - Breast cancer in young women: clinical decision-making in the face of uncertainty. PMID- 19544685 TI - Breast cancer in women under 40. AB - Breast cancer among young women is rare and often associated with high-risk features. Age alone has been shown to be an independent risk factor for recurrence, although the biologic explanation for the increased risk among younger women remains unknown. Due to the higher risk of recurrence and otherwise typically good health of young women with breast cancer, one must carefully consider both the maximally effective approach to treatment and the potential long-term consequences of therapy. This manuscript focuses on the clinical decisions that we confront when considering systemic therapy for young women with early-stage breast cancer and the evolving data to guide the care of these patients. PMID- 19544687 TI - Pregnancy and fertility with breast cancer: what are the options? PMID- 19544688 TI - Complexities of adjuvant endocrine therapy in young premenopausal women. AB - Overall, this excellent overview of management issues for breast cancer patients under 40 years of age highlights the major questions surrounding choices of adjuvant systemic therapy. Treatment choices are dependent on breast cancer biology (receptor status) as well as patient factors (ovarian function and desire for future fertility). As many unanswered questions remain, it is important to continue to enroll subjects in clinical trials focused on this important patient population. PMID- 19544690 TI - The pathway ahead in melanoma trials. PMID- 19544689 TI - Treatment of metastatic melanoma: an overview. AB - The 10-year survival rate for patients with metastatic melanoma is less than 10%. Although surgery and radiation therapy have a role in the treatment of metastatic disease, systemic therapy is the mainstay of treatment for most patients. Single agent chemotherapy is well tolerated but is associated with response rates of only 5% to 20%. Combination chemotherapy and biochemotherapy may improve objective response rates but do not extend survival and are associated with greater toxicity. Immunotherapeutic approaches such as high-dose interleukin-2 are associated with durable responses in a small percentage of patients. In this article, we review the treatments for metastatic melanoma including promising investigational approaches. PMID- 19544691 TI - Treating metastatic melanoma: further considerations. PMID- 19544692 TI - Despite past disappointments, the future of melanoma therapy appears bright. PMID- 19544693 TI - Adjuvant or induction cisplatin-based chemotherapy for operable lung cancer. AB - Despite aggressive surgical management, 5-year survival rates of patients with non-small-cell lung cancer (NSCLC) range from 73% for those with pathologic stage IA to 25% for stage IIIA. Given the low survival rate associated with treatment by surgery alone, numerous trials have investigated the use of induction or adjuvant strategies with chemotherapy or thoracic irradiation, either alone or in combination. A meta-analysis published in 1995 showed that cisplatin-based regimens produce the best adjuvant chemotherapy results in NSCLC patients, and this finding has been validated by three large randomized trials. Neoadjuvant chemotherapy offers theoretical advantages over adjuvant chemotherapy, including improved patient compliance, a smaller primary tumor, and pathologic evaluation of treatment efficacy. However, most large randomized trials of neoadjuvant chemotherapy have failed to show statistically significant results. This article reviews the pros and cons of each strategy, current guidelines, and treatment methods that are being explored. PMID- 19544694 TI - Key questions for perioperative chemotherapy in resectable lung cancer: not pre vs post, but who and what? PMID- 19544695 TI - Adjuvant vs neoadjuvant chemotherapy in resectable NSCLC: is that the real question? PMID- 19544696 TI - Therapeutic options in relapsed or refractory diffuse large B-cell lymphoma. Part 1. current treatment approaches. AB - The addition of rituximab to systemic chemotherapy has improved the response rates, progression-free survival, and overall survival of patients with newly diagnosed diffuse large B-cell lymphoma (DLBCL) compared to chemotherapy alone. In the front-line setting, the use of rituximab is changing the biology and clinical behavior in DLBCL patients who fail to respond or relapse following chemoimmunotherapy. Based on retrospective studies, it is becoming evident that the subset of patients with rituximab/chemotherapy-relapsed/refractory DLBCL represents a different clinical entity with a higher degree of chemotherapy resistance compared to DLBCL patients receiving upfront chemotherapy alone. The backbone of treatment for "sensitive"patients with relapsed/refractory disease continues to be salvage chemotherapy with or without rituximab, followed by high dose chemotherapy and autologous stem cell support. Patients who are not eligible for high-dose chemotherapy may benefit from a growing number of regimens integrating novel agents with promising activity and manageable toxicity. Advances in biotechnology have led to the development of novel biomarkers used for categorization or risk stratification in DLBCL patients. This two-part review summarizes treatment options for patients with relapsed/refractory DLBCL and stresses the emerging therapeutic challenges for patients who were previously exposed to rituximab. PMID- 19544697 TI - Complementary therapies, herbs, and other OTC agents. PMID- 19544698 TI - Eradicating malaria. AB - The renewed interest in malaria research and control is based on the intolerable toll this disease takes on young children and pregnant women in Africa and other vulnerable populations; 150 to 300 children die each hour from malaria amounting to 1 to 2 million deaths yearly. Malaria-induced neurologic impairment, anemia, hypoglycemia, and low birth weight imperil normal development and survival. Resistance of Plasmodium falciparum to drugs and Anopheles mosquitoes to insecticides has stimulated discovery and development of artemisinin-based combination treatments (ACTs) and other drugs, long-lasting insecticide-treated bednets (with synthetic pyrethroids) and a search for non-toxic, long-lasting, affordable insecticides for indoor residual spraying (IRS). Malaria vaccine development and testing are progressing rapidly and a recombinant protein (RTS,S/AS02A) directed against the circumsporozoite protein is soon to be in Phase 3 trials. Support for malaria control, research, and advocacy through the Global Fund for HIV/AIDS, Tuberculosis and Malaria, the U.S. President's Malaria Initiative, the Bill & Melinda Gates Foundation, WHO and other organizations is resulting in decreasing morbidity and mortality in many malarious countries. Sustainability of effective programs through training and institution strengthening will be the key to malaria elimination coupled with improved surveillance and targeted research. PMID- 19544699 TI - Oil from algae; salvation from peak oil? AB - A review is presented of the use of algae principally to produce biodiesel fuel, as a replacement for conventional fuel derived from petroleum. The imperative for such a strategy is that cheap supplies of crude oil will begin to wane within a decade and land-based crops cannot provide more than a small amount of the fuel the world currently uses, even if food production were allowed to be severely compromised. For comparison, if one tonne of biodiesel might be produced say, from rape-seed per hectare, that same area of land might ideally yield 100 tonnes of biodiesel grown from algae. Placed into perspective, the entire world annual petroleum demand which is now provided for by 31 billion barrels of crude oil might instead be met from algae grown on an area equivalent to 4% of that of the United States. As an additional benefit, in contrast to growing crops it is not necessary to use arable land, since pond-systems might be placed anywhere, even in deserts, and since algae grow well on saline water or wastewaters, no additional burden is imposed on freshwater-a significant advantage, as water shortages threaten. Algae offer the further promise that they might provide future food supplies, beyond what can be offered by land-based agriculture to a rising global population. PMID- 19544700 TI - Surveillance of infectious complications in hemato-oncological patients. AB - BACKGROUND: Monitoring the rate of infections in individual centers that treat patients with hematological malignancies is of major importance. However, there are no uniform guidelines for infection surveillance. OBJECTIVES: To describe the epidemiology of bacterial and fungal infections in a single hematology ward and to compare methods for reporting surveillance and infection rates in other centers in Israel. METHODS: We conducted a prospective surveillance of all patients admitted to our hematology ward, applying standard definitions for invasive fungal infections and adapting definitions for non-fungal infections. Incidence rates were calculated using patients, admissions, hospital days and neutropenia days. We performed a search for other reported surveillance studies in Israel. , RESULTS: We detected 79 infectious episodes among 159 patients admitted to the hematology ward during 1 year. Using neutropenia days as the denominator for calculation of incidence discriminated best between patients at high and low risk for infection. The incidence of invasive fungal infections was 7, 10 and 18 per 1000 neutropenia days, among all patients, those with acute leukemia and those with acute leukemia undergoing induction therapy, respectively. Only 10 reports from Israel were identified, 6 of which were prospective. Our data could not be compared to these reports because of the varying definitions and denominators used. CONCLUSIONS: Hematology centers should monitor infection rates and report them in a uniform methodology. PMID- 19544701 TI - Are the "good old" antibiotics still appropriate for early-onset neonatal sepsis? A 10 year survey. AB - BACKGROUND: Early-onset neonatal sepsis is a major cause of morbidity and mortality among newborn infants. OBJECTIVES: To determine the incidence, type of pathogens and resistance to antibiotics among newborns with early-onset neonatal sepsis, and to identify the risk factors predisposing infants to resistant pathogens in order to reevaluate antibiotic regimens appropriate for resistant bacteria in these high risk neonates. METHODS: We retrospectively studied maternal and neonatal variables of 73 term and near-term infants and 30 preterm infants, born over a period of 10.5 years and exhibiting early-onset neonatal sepsis (positive blood cultures in the first 72 hours of life). RESULTS: Predominant pathogens in term and near-term infants were gram-positive compared with gram-negative organisms (mostly Escherichia coli) in preterm infants. Mothers of infants with antibiotic-resistant organisms weremore likely to have prolonged rupture of membranes and prolonged hospitalization before delivery and to be treated with antibiotics. No trends towards more resistant strains of pathogens were recorded over the 10.5 years of the study period. CONCLUSIONS: Early-onset neonatal sepsis in term infants differs in bacterial species from that in preterm infants, with predominantly gram-positive organisms in term and near-term infants and gram-negative organisms in preterms. Rates of bacterial resistance to the combination of ampicillin and gentamicin, though higher among infants born to mothers with prolonged hospitalization who had been treated with antibiotics, still remained very low in our department. Thus, it seems that our classic antibiotic regimen is still appropriate for both term and preterm newborns. PMID- 19544702 TI - MRI, not CT, to rule out recurrent cholesteatoma and avoid unnecessary second look mastoidectomy. AB - BACKGROUND: Aural cholesteatoma is an epidermal cyst of the middle ear or mastoid that can be eradicated only by surgical resection. It is usually managed with radical or modified radical mastoidectomy. Clinical diagnosis of recurrent cholesteatoma in a closed postoperative cavity is difficult. Thus, the accepted protocol in most otologic centers for suspected recurrence consists of second look procedures performed approximately 1 year after the initial surgery. Brain herniation into a post-mastoidectomy cavity is not rare and can be radiologically confused with cholesteatoma on the high resolution computed tomographic images of temporal bones that are carried out before second-look surgery. OBJECTIVES: To present our experience with meningoceles that were confused with recurrent disease in patients who had undergone primary mastoidectomy for cholesteatoma and to support the use of magnetic resonance imaging as more suitable than CT in postoperative follow-up protocols for cholesteatoma. METHODS: We conducted a retrospective chart review of four patients. RESULTS: Axial CT sections demonstrated a soft tissue mass in the middle ear and mastoid in all four patients. Coronal reconstructions of CT scans showed a tympanic tegmen defect in two patients. CT failed to exclude cholesteatoma in any patient. Each underwent a second-look mastoidectomy and the only finding at surgery was meningocele in all four patients. CONCLUSIONS: Echo-planar diffusion-weighted MRI can differentiate between brain tissue and cholesteatoma more accurately than CT. We recommend that otolaryngologists avoid unnecessary revision procedures by using the newest imaging modalities for more precise diagnosis of patients who had undergone mastoidectomy for cholesteatoma in the past. PMID- 19544703 TI - QT interval disturbances in hospitalized elderly patients. AB - BACKGROUND: The QT interval reflects the total duration of ventricular myocardial repolarization. Its prolongation is associated with increased risk of polymorphic ventricular tachycardia, or torsade de pointes, which can be fatal. OBJECTIVES: To assess the prevalence of both prolonged and short QT interval in patients admitted to an acute geriatric ward. METHODS: This retrospective study included the records over 6 months of all patients hospitalized in an acute geriatric ward. Excluded were patients with pacemaker, bundle branch block and slow or rapid atrial fibrillation. The standard 12 lead electrocardiogram of each patient was used for the QT interval evaluation. RESULTS: We screened the files of 422 patients. QTc prolongation based on the mean of 12 ECG leads was detected in 115 patients (27%). Based on lead L2 only, QTc was prolonged in 136 (32%). Associated factors with QT prolongation were congestive heart failure and use of hypnotics. Short QTwas found in 30 patients (7.1%) in lead L2 and in 19 (4.5%) by the mean 12 leads. Short QT was related to a higher heart rate, chronic atrial fibrillation and schizophrenia. CONCLUSIONS: Our study detected QT segment disturbances in a considerable number of elderly patients admitted acutely to hospital. Further studies should confirm these results and clinicians should consider a close QT interval follow-up in predisposed patients. PMID- 19544704 TI - Hyperimmune gammaglobulin for the treatment of West Nile virus encephalitis. AB - BACKGROUND: West Nile virus, the etiologic agent of West Nile fever, is an emerging mosquito-borne disease. WNV was recognized as a cause of severe human meningoencephalitis in elderly patients during outbreaks in various parts of the world. OBJECTIVES: To analyze WNV encephalitis therapy and its outcome after prescribing hyperimmune gammaglobulin therapy. METHODS: Eight subjects with WNV encephalitis were treated with supportive therapy and 5 days of IVIG 0.4 g/kg/day containing high WNV antibodies obtained from healthy blood donors. RESULTS: Patients who were treated with IVIG as soon as possible exhibited an improvement in their symptoms. All subjects presented with high fever, progressive confusion and headaches, nausea and vomiting. The Glasgow Coma Screen for six patients ranged between 8 and 13 and all were discharged with a score of 15. The remaining two subjects died during their hospitalization. CONCLUSIONS: In severe WNV infection, where the disease affects the central and/or peripheral nervous system, early intervention with IVIG together with supportive treatment is recommended. PMID- 19544705 TI - Clinical results of unprotected left main coronary stenting. AB - BACKGROUND: Although unprotected left main coronary artery disease is considered by contemporary guidelines to be an indication for surgery, percutaneous coronary intervention may be necessary in patients at high surgical risk. OBJECTIVES: To assess the outcome of angioplasty in the treatment of unprotected LMCA disease. METHODS: Angiographic and clinical data were collected prospectively for all patients who underwent emergent or nonemergent (planned) therapeutic PCI for unprotected LMCA disease at our center from 2003 to 2007. Baseline values were compared with findings at 1, 6 and 12 months after the procedure. RESULTS: The study group comprised 71 consecutive patients with a mean age of 74 +/- 12 years; 63% were men, and 31% had diabetes. Forty-three patients had a planned procedure and 28 an emergent procedure. Mean EuroScore was 7.3 +/- 3.6 (range 5-12). Forty nine percent of the procedures were performed with bare metal stents and 51% with drug-eluting stents. Procedural success was achieved in 100% of cases. The overall mortality rate was 11.3% at 1 month, 18.3% at 6 months and 19.7% at 12 months. Elective PCI was associated with significantly lower mortality (2.3% vs. 25% at 1 month, 4.6% vs. 39% at 6 months and 6.9% vs. 39% at 12 months), and the use of drug-eluting stents was associated with lower rates of target vessel revascularization and major adverse cardiac events than use of bare metal stents (2.8% vs. 14% at 1 month, 8.3% vs. 43% at 6 and 12 months). Variables that correlated with increased mortality or MACE at 6 and 12 months were cardiogenic shock, emergent PCI, ejection fraction < 35%, renal failure, distal left main stenosis location, and reference diameter < 3 mm. CONCLUSIONS: PCI is a feasible and relatively safe therapeutic option for unprotected LMCA. The less favorable outcome of emergent compared to planned PCI is probably attributable to the overwhelming acute myocardial ischemic injury in emergent cases. The use of drug eluting stents may improve the intermediate-term restenosis rate. PMID- 19544706 TI - Vision screening among northern Israeli Jewish and Arab schoolchildren. AB - BACKGROUND: Uncorrected refractive error is the leading cause of visual impairment in children. In 2002 a screening project was launched in Israel to provide data on the effectiveness of the illiterate E-chart in identifying Jewish and Arab schoolchildren in need of a comprehensive eye examination. OBJECTIVES: To present the aims, design and initial results of the visual screening project and the prevalence of vision abnormality in the study population. METHODS: A cross-sectional population-based study was conducted during 2002-2003 among first and eighth-graders in 70 schools in northern Israel. The nurse's test included use of the illiterate E-chart to measure visual acuity. The medical examination included vision history, clinical eye examination, VA and retinoscopy. The ophthalmologist's evaluation as to whether a child needed a referral for, diagnostic procedures, treatment and/or follow-up was recorded and compared with explicit referral criteria formulated after data collection. RESULTS: Of 1975 schoolchildren, 31% had abnormal VA, defined as VA worse than 6/6 in at least one eye, and a quarter had VA equal or worse than 6/12 in both eyes. The prevalence of vision abnormality among the children was 22.4% when based on the evaluation of the field ophthalmologist and 26.1% when based on two sets of explicit severity scores and referral criteria. CONCLUSIONS: Vision abnormality is a significant health problem among northern Israeli schoolchildren. This project is unique in scope and importance, providing evidence to assist policy making with regard to vision screening for schoolchildren (including data on test reliability and validity) and optimal VA cutoff level, and confirming the need for clinical guidelines regarding referral criteria. PMID- 19544707 TI - Endoscopic ultrasound: doubtful accuracy for restaging esophageal cancer after preoperative chemotherapy. AB - BACKGROUND: The role of endoscopic ultrasound in evaluating the response of esophageal cancer to neoadjuvant chemotherapy is controversial. OBJECTIVES: To evaluate the accuracy of EUS in restaging patients who underwent NAC. METHODS: The disease stage of patients with esophageal cancer was established by means of the TNM classification system. The initial staging was determined by chest and abdominal computed tomography and EUS. Patients who needed NAC underwent a preoperative regimen consisting of cisplatin and fluouracil. Upon completion of the chemotherapy, patients were restaged and then underwent esophagectomy. The results of the EUS staging were compared with the results of the surgical pathology staging. This comparison was done in two groups of patients: the study group (all patients who received NAC) and the control group (all patients who underwent primary esophagectomy without NAC). RESULTS: NAC was conducted in 20 patients with initial stage IIB and III carcinoma of the esophagus (study group). Post-chemotherapy EUS accurately predicted the surgical pathology stage in 6 patients (30%). Pathological down-staging was noted in 8 patients (40%). However, the EUS was able to observe it in only 2 patients (25%). The accuracy of EUS in determining the T status alone was 80%. The accuracy for N status alone was 35%. In 65% of examinations the EUS either overestimated (35%) or underestimated (30%) the N status. Thirteen patients with initial stage I-IIA underwent primary esophagectomy after the initial staging (control group). EUS accurately predicted the surgical pathology disease stage in 11 patients (85%). CONCLUSIONS: EUS is an accurate modality for initial staging of esophageal carcinoma. However, it is not a reliable tool for restaging esophageal cancer after NAC and it cannot predict response to chemotherapy. PMID- 19544708 TI - Elastofibroma at the scapular region. AB - BACKGROUND: Elastofibroma is a rare type of lesion consisting of elastic fibers within a stroma of collagen and fatty tissue. It is usually located on the lower scapular region attached firmly to the thoracic cage, often causing debilitating pain. Its clinical presentation mimics a soft tissue tumor. OBJECTIVES: To evaluate the diagnosis and treatment results of elastofibroma. METHODS: Clinical and radiographic evaluations were performed in 11 patients with thoracic wall mass. In five of them a biopsy was taken before surgery. All patients were operated and the diagnosis of elastofibroma was confirmed by histology. RESULTS: Two patients had a postoperative seroma that resolved spontaneously within a few days. All patients reesumed their preoperative activities, including sports. CONCLUSIONS: Considering the slow-growing nature of this tumor and its typical presentation, we believe that when this diagnosis is suspected, investigation does not necessitate staging (as in sarcomas). Also, marginal surgical excision is sufficient. Observation is an acceptable alternative to surgery. PMID- 19544709 TI - Angiographic evaluation of epicardial and microvascular coronary flow. PMID- 19544710 TI - Aspirin in recurrent miscarriage: is there an indication? PMID- 19544711 TI - Influenza vaccine and autoimmunity. PMID- 19544712 TI - Ischemic stroke in a patient with lupus following influenza vaccination: a questionable association . PMID- 19544713 TI - Clinical cross-reactivity between danaparoid and heparin antibodies successfully managed with bivalirudin. PMID- 19544715 TI - Stress: a dilemma concerning healthy CJD mutation carriers. PMID- 19544714 TI - Cervical thymic cyst: unusual age and site. PMID- 19544716 TI - [Is the "psychopathy concept" applicable to expert opinions in civil suits?]. AB - The assessment of the psychological consequences of an accident is a complex and sophisticated challenge for physicians. Pre-traumatic personality traits and the post-traumatic course must be assessed with care. The evaluation of pre-traumatic personality traits is often an issue of particular debate. In this paper, the concept of "psychopathy" according to Hare is outlined, and the potential application of this concept in the assessment procedure is discussed. Typical personality traits such as glibness, superficial charm, egocentricity, pathological lying and deception, lack of remorse and lack of empathy may provide valuable hints for the expert opinion. PMID- 19544717 TI - [Workplace-related anxiety, workplace phobia and disorders of participation]. AB - Work is an important domain of life. It is therefore clear that problems at the workplace and mental disorders will have negative interactions. Job-related anxieties are of special importance as any workplace causes or intensifies anxiety by its very nature. A common final pathway of mental disorders in general and workplace-related anxieties in particular is workplace phobia. Similarly to agoraphobia, it is characterised by panic when approaching or even thinking of the stimulus, in this case the workplace. Workplace phobia has serious negative consequences for the further course of illness. It impairs the ability to work, and can lead to sick leave and early retirement. It requires special therapeutic interventions. This paper describes workplace-related anxieties and workplace phobia and gives a conceptual framework for their understanding. PMID- 19544718 TI - [The "Interview for evaluation of the psychosocial dimensions of capability performance"--a systematic add-on instrument for the expert assessment of work capacity]. AB - The diagnosis of psychiatric or psychosomatic disorders as such does not represent a complete indicator of the work capacity of an individual. Further sociomedical patterns need to be recognised for an expert assessment of capability. As far as we know, no scientifically accepted diagnostic instrument for work-capacity testimonials currently exists that also appreciates psychosocial dimensions. The "Interview for the evaluation of the psychosocial dimensions of capability performance" represents a potentially suitable instrument for the assessment of capability performance in various contexts. It is highly viable in terms of the time required and can be applied without special training on the content. PMID- 19544719 TI - [Quality assurance by routine data (QAR)--a new dimension in quality management of inpatient treatment?]. AB - Patients, admitting doctors and payers want to have more and more valid information about the medical results in hospitals really are. The quality of medical care in hospitals is demonstrated in the quality reports through structure and process data, data of volume about the most common main diagnoses and procedures and, in the current version, also through data of external quality assurance. For patients and payers, this means that it may be difficult to obtain sufficient information on relevant quality criteria for different therapies in hospital departments. It may also mean that it is difficult to choose between hospitals. The project "Quality assurance of inpatient treatment by routine data" (QAR) is a new and extended attempt in quality management. By changing the reimbursement procedures of hospitals to DRG, one receives data from which one can easily deduce marks of quality. For special treatment the QAR marks of quality provide patients, insurance companies and the internal quality management of hospitals with information on the quality of the hospitals and their departments. In this paper, the opportunities and prospects of quality assurance by using routine date will be discussed. PMID- 19544720 TI - [Philosophy of the mutual biotic system of man-environment]. AB - With regard to environmental changes, outstanding importance is meanwhile to be attached to the cultural side of human evolution. The evolution both of mankind and of its environment are mutually dependent as processes of change and together they form a complete biotic system. First disorders of balance concerning the close relationship network between mankind and environment eventually developed following man's change from the biosphere to the "noosphere" created by him. In the course of the "neolithic revolution" mankind, while becoming more and more settled, began to become increasingly estranged from its ecological surroundings. Environmental problems caused by man led to climatic changes already about 8,000 years ago. So far they have caused an extraordinary climatic stability following the Ice Age. "Environmental art" i. e. an improved evolution - is required to escape an imminent "collapse" caused by pollution. Nowadays mankind is on the way to being the almost exclusive carrier of future evolution of this planet. PMID- 19544721 TI - An influential reach. Nurses have different roles, but all seek to improve practice, workplace. PMID- 19544722 TI - We put safety first. PMID- 19544723 TI - Funding for public health services tightens as need rises. PMID- 19544725 TI - University program supports students, builds diverse nursing workforce. PMID- 19544724 TI - ANA program goes mobile to keep nurses safe from chemical substances. PMID- 19544726 TI - Where and when to revegetate: a quantitative method for scheduling landscape reconstruction. AB - Restoration of native vegetation is required in many regions of the world, but determining priority locations for revegetation is a complex problem. We consider the problem of determining spatial and temporal priorities for revegetation to maximize habitat for 62 bird species within a heavily cleared agricultural region, 11000 km2 in area. We show how a reserve-selection framework can be applied to a complex, large-scale restoration-planning problem to account for multi-species objectives and connectivity requirements at a spatial extent and resolution relevant to management. Our approach explicitly accounts for time lags in planting and development of habitat resources, which is intended to avoid future population bottlenecks caused by delayed provision of critical resources, such as tree hollows. We coupled species-specific models of expected habitat quality and fragmentation effects with the dynamics of habitat suitability following replanting to produce species-specific maps for future times. Spatial priorities for restoration were determined by ranking locations (150-m grid cells) by their expected contribution to species habitat through time using the conservation planning tool, "Zonation." We evaluated solutions by calculating expected trajectories of habitat availability for each species. We produced a spatially explicit revegetation schedule for the region that resulted in a balanced increase in habitat for all species. Priority areas for revegetation generally were clustered around existing vegetation, although not always. Areas on richer soils and with high rainfall were more highly ranked, reflecting their potential to support high-quality habitats that have been disproportionately cleared for agriculture. Accounting for delayed development of habitat resources altered the rank-order of locations in the derived revegetation plan and led to improved expected outcomes for fragmentation-sensitive species. This work demonstrates the potential for systematic restoration planning at large scales that accounts for multiple objectives, which is urgently needed by land and natural resource managers. PMID- 19544727 TI - Growth and nutritional state of American Crow nestlings vary between urban and rural habitats. AB - In urbanized areas, many adult birds find sufficient foods to survive, but the anthropogenic foods that are abundant there may be detrimental to nestling growth. In fact, American Crow (Corvus brachyrhynchos) nestlings are smaller in suburban than rural areas, possibly because of nutrient limitation. Here, we seek to identify possible causes of size differences by comparing both size and blood chemistry measures in rural and suburban crow nestlings. We quantified land use in known crow territories and distinguished three distinct environments: suburban residential, suburban-managed (e.g., golf courses), and rural. We measured nestlings near fledging age in each environment and bled them for determination of unbound plasma calcium, total protein, and corticosterone. We supplemented a subset of broods in suburban-residential and rural areas with a food high in protein and calcium. Rural nestlings were significantly larger than suburban residential crows and had higher total serum protein. Nestlings in suburban managed areas were intermediate in size and serum protein but had the lowest plasma calcium levels. Nestling corticosterone levels did not differ significantly among habitats, indicating that, although suburban nestlings may be food-limited, they were not starving. Supplemented nestlings in suburban residential areas were significantly larger in some growth measures than their unsupplemented counterparts. Unexpectedly, supplemented rural nestlings were significantly smaller than unsupplemented rural ones, suggesting that parents use easily accessible food even when it is nutritionally suboptimal. Our results indicate that nestlings in suburban areas are nutrient restricted, rather than calorie restricted. PMID- 19544728 TI - Goats, birds, and emergent diseases: apparent and hidden effects of exotic species in an island environment. AB - Exotic species can have devastating effects on recipient environments and even lead to the outbreak of emergent diseases. We present here several hidden effects that the introduction of goats has had on the Lesser Short-toed Lark, Calandrella rufescens, the commonest native bird inhabiting the island of Fuerteventura (Canary Islands). Vegetation structure varied with grazing pressure, and indeed, vegetation was all but eradicated from the locality with greatest goat densities, which was also where the lowest density of Lesser Short-toed Larks was recorded. The impact of habitat impoverishment, however, was partially compensated for by changes in the foraging behavior of birds, which benefited from the abundant food provided to goats on farms. Capture-resighting methods showed that birds visiting farms outnumbered the estimates for birds obtained in the surrounding natural habitat, suggesting that there was recruitment from a much larger area. Stable isotope analyses of feathers indicated that island birds feed largely on the maize supplied at goat farms, showing poorer body condition than birds from populations not associated with farms (peninsular Spain and Morocco). Moreover, larks from Fuerteventura had a very high prevalence of poxvirus lesions compared with other bird populations worldwide and may increase the risk of contracting the disease by feeding on farms, where they aggregate and coexist atypically with domestic birds. The island birds also had lower average productivity, which may be the consequence of the emergent disease and/or the poor nutritional state resulting from feeding on a low-protein diet. Diseased and non-diseased birds from Fuerteventura showed similar body condition and annual survival rates. However, the isotopic traces of delta 13C indicate that the diet of diseased birds was more uniform than that of non-diseased birds, being based on food from goat farms. Our results show how the combination of species frequently introduced onto islands (goats, poultry, and associated pathogens) can create ecological traps for native species that are not always easy to identify. Moreover, we stress that nutrition and infectious diseases are important determinants of the well-being and dynamics of animal populations, and thus health research must be included in the design of monitoring programs and conservation strategies. PMID- 19544729 TI - Bushmeat poaching reduces the seed dispersal and population growth rate of a mammal-dispersed tree. AB - Myriad tropical vertebrates are threatened by overharvest. Whether this harvest has indirect effects on nonhunted organisms that interact with the game species is a critical question. Many tropical birds and mammals disperse seeds. Their overhunting in forests can cause zoochorous trees to suffer from reduced seed dispersal. Yet how these reductions in seed dispersal influence tree abundance and population dynamics remains unclear. Reproductive parameters in long-lived organisms often have very low elasticities; indeed the demographic importance of seed dispersal is an open question. We asked how variation in hunting pressure across four national parks with seasonal forest in northern Thailand influenced the relative abundance of gibbons, muntjac deer, and sambar deer, the sole dispersers of seeds of the canopy tree Choerospondias axillaris. We quantified how variation in disperser numbers affected C. axillaris seed dispersal and seedling abundance across the four parks. We then used these data in a structured population model based on vital rates measured in Khao Yai National Park (where poaching pressure is minimal) to explore how variation in illegal hunting pressure might influence C. axillaris population growth and persistence. Densities of the mammals varied strongly across the parks, from relatively high in Khao Yai to essentially zero in Doi Suthep-Pui. Levels of C. axillaris seed dispersal and seedling abundance positively tracked mammal density. If hunting in Khao Yai were to increase to the levels seen in the other parks, C. axillaris population growth rate would decline, but only slightly. Extinction of C. axillaris is a real possibility, but may take many decades. Recent and ongoing extirpations of vertebrates in many tropical forests could be creating an extinction debt for zoochorous trees whose vulnerability is belied by their current abundance. PMID- 19544730 TI - The costs of anti-herbivore defense traits in agricultural crop plants: a case study involving leafhoppers and trichomes. AB - The expression of plant defenses is thought to entail costs (e.g., the allocation of resources away from growth or reproduction) that constrain the evolution of plant genotypes maximally defended against herbivores. Although central to the ecological theory underlying plant-insect interactions at large, the concept of defense costs is particularly evident in agricultural crops where plants may be under simultaneous selection for enhanced growth and/or reproduction (i.e., yield) and anti-herbivore resistance traits that deter pests. In this study we investigate the role of trichomes as a resistance mechanism against a sap-feeding insect (the leafhopper, Empoasca fabae) on potato. Natural variation in trichome density among 17 potato cultivars was used to test for the role of trichomes as a putative defense against leafhoppers, and evidence of costs in trichome expression. Two different types of costs were explored: (1) allocation costs (i.e., the relationship between trichomes and yield), and (2) costs involving trade-offs with alternative defense strategies (e.g., tolerance). Although leafhopper abundance did not decrease as trichome density increased, leafhopper injury to potato plants (foliar necrosis) was negatively correlated with trichome density. As a result, the per capita effect of leafhopper adults and nymphs on foliar damage was lower on plants with high trichome densities. We found no evidence, however, for costs of expressing this resistance trait; trichomes were not correlated with either potato yield or tolerance to herbivory. Thus, selection for multiple plant defenses to alleviate the impact of pests in agronomic crops may indeed be possible without inherent losses in plant yield. PMID- 19544731 TI - Ecological, behavioral, and genetic factors influencing the recombinant control of invasive pests. AB - Invasive species are a major threat to biodiversity, cost the world economy billions of dollars annually, and are often difficult, if not impossible, to control using current approaches. Recombinant technologies could revolutionize management of such pests but would be subject to a range of genetic, behavioral, and ecological factors that could limit their efficacy or applicability. We use a realistically parameterized combined population dynamics/genetics model to assess the potential of, and constraints on, a suite of recombinant approaches that have been suggested for pest control. We show that, of the options suggested to date, a genetic construct that distorts operational sex ratios by sterilizing, killing, or sex-changing one gender and being inherited through the other, is not only potentially the most effective means of pest control, but also one that remains effective over the widest range of ecological and behavioral conditions. All methods, however, are sensitive in particular to the degree of density dependence in the pest population and to operational issues such as maximum copy number and stocking levels, which affect introgression rates. Optimal investment strategies for an integrated pest management program that includes the nonlinear interactions of recombinant strategies and complementary management options can be assessed through the sensitivity analyses. The subtle effects of even minor variability in some parameters, such as extra mortality due to the presence of the construct, further suggest that genetic techniques be applied in an active adaptive management framework, so that strategies can be regularly optimized as the impacts of a release program are assessed. PMID- 19544732 TI - Development and application of DNA techniques for validating and improving pinniped diet estimates. AB - Polymerase chain reaction techniques were developed and applied to identify DNA from >40 species of prey contained in fecal (scat) soft-part matrix collected at terrestrial sites used by Steller sea lions (Eumetopias jubatus) in British Columbia and the eastern Aleutian Islands, Alaska. Sixty percent more fish and cephalopod prey were identified by morphological analyses of hard parts compared with DNA analysis of soft parts (hard parts identified higher relative proportions of Ammodytes sp., Cottidae, and certain Gadidae). DNA identified 213 prey occurrences, of which 75 (35%) were undetected by hard parts (mainly Salmonidae, Pleuronectidae, Elasmobranchii, and Cephalopoda), and thereby increased species occurrences by 22% overall and species richness in 44% of cases (when comparing 110 scats that amplified prey DNA). Prey composition was identical within only 20% of scats. Overall, diet composition derived from both identification techniques combined did not differ significantly from hard-part identification alone, suggesting that past scat-based diet studies have not missed major dietary components. However, significant differences in relative diet contributions across scats (as identified using the two techniques separately) reflect passage rate differences between hard and soft digesta material and highlight certain hypothesized limitations in conventional morphological-based methods (e.g., differences in resistance to digestion, hard part regurgitation, partial and secondary prey consumption), as well as potential technical issues (e.g., resolution of primer efficiency and sensitivity and scat subsampling protocols). DNA analysis of salmon occurrence (from scat soft-part matrix and 238 archived salmon hard parts) provided species-level taxonomic resolution that could not be obtained by morphological identification and showed that Steller sea lions were primarily consuming pink (Oncorhynchus gorbuscha) and chum (Oncorhynchus keta) salmon. Notably, DNA from Atlantic salmon (Salmo salar) that likely originated from a distant fish farm was also detected in two scats from one site in the eastern Aleutian Islands. Overall, molecular techniques are valuable for identifying prey in the fecal remains of marine predators. Combining DNA and hard-part identification will effectively alleviate certain predicted biases and will ultimately enhance measures of diet richness, fisheries interactions (especially salmon-related ones), and the ecological role of pinnipeds and other marine predators, to the benefit of marine wildlife conservationists and fisheries managers. PMID- 19544733 TI - Fishing from past to present: continuity and resilience of red abalone fisheries on the Channel Islands, California. AB - Archaeological data from coastal shell middens provide a window into the structure of ancient marine ecosystems and the nature of human impacts on fisheries that often span millennia. For decades Channel Island archaeologists have studied Middle Holocene shell middens visually dominated by large and often whole shells of the red abalone (Haliotis rufescens). Here we use modern ecological data, historical accounts, commercial red abalone catch records, and zooarchaeological data to examine long-term spatial and temporal variation in the productivity of red abalone fisheries on the Northern Channel Islands, California (USA). Historical patterns of abundance, in which red abalone densities increase from east to west through the islands, extend deep into the Holocene. The correlation of historical and archaeological data argue for long-term spatial continuity in productive red abalone fisheries and a resilience of abalone populations despite dramatic ecological changes and intensive human predation spanning more than 8000 years. Archaeological, historical, and ecological data suggest that California kelp forests and red abalone populations are structured by a complex combination of top-down and bottom-up controls. PMID- 19544734 TI - Mapping the bycatch seascape: multispecies and multi-scale spatial patterns of fisheries bycatch. AB - Fisheries bycatch is a worldwide conservation issue. Despite a growing awareness of bycatch problems in particular ocean regions, there have been few efforts to identify spatial patterns in bycatch events. Furthermore, many studies of fisheries bycatch have been myopic, focusing on a single species or a single region. Using a range of analytical approaches to identify spatial patterns in bycatch data, we demonstrate the utility and applications of area and point pattern analyses to single and multispecies bycatch seascapes of pelagic longline fisheries in the Atlantic and Pacific Oceans. We find clear evidence of spatial clustering within bycatch species in both ocean basins, both in terms of the underlying pattern of the locations of bycatch events relative to fishing locations and for areas of high bycatch rates. Furthermore, we find significant spatial overlap in the pattern of bycatch across species relative to the spatial distribution in fishing effort and target catch. These results point to the importance of considering spatial patterns of both single and multispecies bycatch to meet the ultimate goal of reducing bycatch encounters. These analyses also highlight the importance of considering bycatch relative to target catch as a way of identifying areas where fishing effort reduction may help to reduce multispecies bycatch with minimal impact on target catch. PMID- 19544735 TI - Effectiveness of engineered in-stream structure mitigation measures to increase salmonid abundance: a systematic review. AB - Engineered in-stream structures are often installed to increase salmonid abundance, either for commercial gain in fisheries or for conservation purposes in degraded habitats. Having been in widespread use for the last 80 years, at an estimated cost of hundreds of millions of U.S. dollars each year, the effectiveness of these structures is still widely debated in the literature. Many studies of varying quality have been undertaken that attempt to address this issue, but it has proved difficult for practitioners to develop a consensus regarding the utility of these structures, despite their continued use. Systematic review methodology was used to formally synthesize empirical evidence regarding the effectiveness of engineered in-stream structures as a management tool to increase salmonid abundance. Meta-analysis shows that evidence regarding the effectiveness of in-stream devices is equivocal. Heterogeneity is significant both for population size and local habitat preference. This heterogeneity is related to stream width, with in-stream devices being less effective in larger streams. Consequently, widespread use of in-stream structures for restoration, particularly in larger streams, is not supported by the current scientific evidence base. PMID- 19544736 TI - Effects of agricultural subsidies of nutrients and detritus on fish and plankton of shallow-reservoir ecosystems. AB - Agricultural activities increase exports of nutrients and sediments to lakes, with multiple potential impacts on recipient ecosystems. Nutrient inputs enhance phytoplankton and upper trophic levels, and sediment inputs can shade phytoplankton, interfere with feeding of consumers, and degrade benthic habitats. Allochthonous sediments are also a potential food source for detritivores, as is sedimenting autochthonous phytodetritus, the production of which is stimulated by nutrient inputs. We examined effects of allochthonous nutrient and sediment subsidies on fish and plankton, with special emphasis on gizzard shad (Dorosoma cepedianum). This widespread and abundant omnivorous fish has many impacts on reservoir ecosystems, including negative effects on water quality via nutrient cycling and on fisheries via competition with sportfish. Gizzard shad are most abundant in agriculturally impacted, eutrophic systems; thus, agricultural subsidies may affect reservoir food webs directly and by enhancing gizzard shad biomass. We simulated agricultural subsidies of nutrients and sediment detritus by manipulating dissolved nutrients and allochthonous detritus in a 2 x 2 factorial design in experimental ponds. Addition of nutrients alone increased primary production and biomass of zooplanktivorous fish (bluegill and young-of year gizzard shad). Addition of allochthonous sediments alone increased algal sedimentation and decreased seston and sediment C:P ratios. Ponds receiving both nutrients and sediments showed highest levels of phytoplankton and total phosphorus. Adult and juvenile gizzard shad biomass was enhanced equally by nutrient or sediment addition, probably because this apparently P-limited detritivore ingested similar amounts of P in all subsidy treatments. Nutrient excretion rates of gizzard shad were higher in ponds with nutrient additions, where sediments were composed mainly of phytodetritus. Therefore, gizzard shad can magnify the direct effects of nutrient subsidies on phytoplankton production, and these multiple effects must be considered in strategies to manage eutrophication and fisheries in warmwater reservoir lakes where gizzard shad can dominate fish biomass. PMID- 19544737 TI - Assessment of lake sensitivity to acidic deposition in national parks of the Rocky Mountains. AB - The sensitivity of high-elevation lakes to acidic deposition was evaluated in five national parks of the Rocky Mountains based on statistical relations between lake acid-neutralizing capacity concentrations and basin characteristics. Acid neutralizing capacity (ANC) of 151 lakes sampled during synoptic surveys and basin-characteristic information derived from geographic information system (GIS) data sets were used to calibrate the statistical models. The explanatory basin variables that were considered included topographic parameters, bedrock type, and vegetation type. A logistic regression model was developed, and modeling results were cross-validated through lake sampling during fall 2004 at 58 lakes. The model was applied to lake basins greater than 1 ha in area in Glacier National Park (n = 244 lakes), Grand Teton National Park (n = 106 lakes), Great Sand Dunes National Park and Preserve (n = 11 lakes), Rocky Mountain National Park (n = 114 lakes), and Yellowstone National Park (n = 294 lakes). Lakes that had a high probability of having an ANC concentration <100 microeq/L, and therefore sensitive to acidic deposition, are located in basins with elevations >3000 m, with <30% of the catchment having northeast aspect and with >80% of the catchment bedrock having low buffering capacity. The modeling results indicate that the most sensitive lakes are located in Rocky Mountain National Park and Grand Teton National Park. This technique for evaluating the lake sensitivity to acidic deposition is useful for designing long-term monitoring plans and is potentially transferable to other remote mountain areas of the United States and the world. PMID- 19544738 TI - Anthropogenic modification of the nitrogen cycling within the Greater Hangzhou Area system, China. AB - Based on the mass balance approach, a detailed quantification of nitrogen (N) cycling was constructed for an urban-rural complex system, named the Greater Hangzhou Area (GHA) system, for this paper. The GHA is located in the humid climatic region on the southeastern coast of China, one of the earliest regions in the Yangtze Delta to experience economic development. Total N input into the GHA was calculated at 274.66 Gg/yr (1 Gg = 10(9) g), and total output was calculated at 227.33 Gg/yr, while N accumulation was assessed at 47.33 Gg/yr (17.2% of the total N input). Human activity resulted in 73%of N input by means of synthetic fertilizers, human food, animal feed, imported N containing chemicals, fossil fuel combustion, and other items. More than 69.3% of N was released into the atmosphere, and riverine N export accounted for 22.2% of total N output. N input and output to and from the GHA in 1980 were estimated at 119.53 Gg/yr and 98.30 Gg/yr, respectively, with an increase of 130% and 131%, respectively, during a 24-year period (from 1980 to 2004). The N input increase was influenced by synthetic fertilizers (138%), animal feed (225%), N-containing chemicals (371%), riverine input (311%), and N deposition (441%). Compared to the N balance seen in the arid Central Arizona-Phoenix (CAP) system in the United States, the proportion of N transferred to water bodies in the humid GHA system was found to be 36 times higher than the CAP system. Anthropogenic activity, as it typically does, enhanced the flux of N biogeochemistry in the GHA; however, a lack of an N remover (N pollutant treatment facilities) causes excess reactive N (Nr; such as NH3, N2O, NOx), polluting water bodies and the atmosphere within the GHA. Therefore many challenges remain ahead in order to achieve sustainable development in the rapidly developing GHA system. PMID- 19544739 TI - Anthropogenic and climate influences on biogeochemical dynamics and molecular level speciation of soil sulfur. AB - The soil environment is a primary component of the global biogeochemical sulfur (S) cycle, acting as a source and sink of various S species and mediating oxidation state changes. However, ecological significance of the various S forms and the impacts of human intervention and climate on the amount and structural composition of these compounds are still poorly understood. We investigated the long-term influences of anthropogenically mediated transitions from natural to managed ecosystems on molecular-level speciation, biogeochemical dynamics, and the apparent temperature sensitivity of S moieties in temperate, subtropical, and tropical environments with mean annual temperature (MAT) ranging from 5 degrees C to 21 degrees C, using elemental analysis and X-ray absorption near-edge structure (XANES) spectroscopy. Land-use and land-cover changes led to the depletion of total soil S in all three ecoregions over a period of up to 103 years. The largest decline occurred from tropical forest agroecosystems (67% Kakamega and 76% Nandi, Kenya), compared to losses from temperate (36% at Lethbridge, Canada, and 40% at Pendleton, USA) and subtropical (48% at South Africa) grassland agroecosystems. The total S losses correlated significantly with MAT. Anthropogenic interventions profoundly altered the molecular-level composition and resulted in an apparent shift in oxidation states of organic S from native ecosystems composed primarily of S moieties in intermediate and highly reduced oxidation states toward managed agroecosystems dominated by organic S rich in strongly oxidized functionalities. The most prominent change occurred in thiols and sulfides, the proportion of which decreased by 46% (Lethbridge) and 57% (Pendleton) in temperate agroecosystems, by 46% in subtropical agroecosystems, and by 79% (Nandi) and 81% (Kakamega) in tropical agroecosystems. The proportion of organic S directly linked to O increased by 81%, 168%, 40%, 92%, and 85%, respectively. Among the various organic S functionalities, thiols and sulfides seem to have higher apparent temperature sensitivity, and thus these organic S moieties may become prone to losses due to land-use changes, even from the cooler regions of the world if MAT of these regions rise in the future. PMID- 19544740 TI - Climate and wildfire area burned in western U.S. ecoprovinces, 1916-2003. AB - The purpose of this paper is to quantify climatic controls on the area burned by fire in different vegetation types in the western United States. We demonstrate that wildfire area burned (WFAB) in the American West was controlled by climate during the 20th century (1916-2003). Persistent ecosystem-specific correlations between climate and WFAB are grouped by vegetation type (ecoprovinces). Most mountainous ecoprovinces exhibit strong year-of-fire relationships with low precipitation, low Palmer drought severity index (PDSI), and high temperature. Grass- and shrub-dominated ecoprovinces had positive relationships with antecedent precipitation or PDSI. For 1977-2003, a few climate variables explain 33-87% (mean = 64%) of WFAB, indicating strong linkages between climate and area burned. For 1916-2003, the relationships are weaker, but climate explained 25-57% (mean = 39%) of the variability. The variance in WFAB is proportional to the mean squared for different data sets at different spatial scales. The importance of antecedent climate (summer drought in forested ecosystems and antecedent winter precipitation in shrub and grassland ecosystems) indicates that the mechanism behind the observed fire-climate relationships is climatic preconditioning of large areas of low fuel moisture via drying of existing fuels or fuel production and drying. The impacts of climate change on fire regimes will therefore vary with the relative energy or water limitations of ecosystems. Ecoprovinces proved a useful compromise between ecologically imprecise state-level and localized gridded fire data. The differences in climate-fire relationships among the ecoprovinces underscore the need to consider ecological context (vegetation, fuels, and seasonal climate) to identify specific climate drivers of WFAB. Despite the possible influence of fire suppression, exclusion, and fuel treatment, WFAB is still substantially controlled by climate. The implications for planning and management are that future WFAB and adaptation to climate change will likely depend on ecosystem-specific, seasonal variation in climate. In fuel limited ecosystems, fuel treatments can probably mitigate fire vulnerability and increase resilience more readily than in climate-limited ecosystems, in which large severe fires under extreme weather conditions will continue to account for most area burned. PMID- 19544741 TI - Changes in vegetation in northern Alaska under scenarios of climate change, 2003 2100: implications for climate feedbacks. AB - Assessing potential future changes in arctic and boreal plant species productivity, ecosystem composition, and canopy complexity is essential for understanding environmental responses under expected altered climate forcing. We examined potential changes in the dominant plant functional types (PFTs) of the sedge tundra, shrub tundra, and boreal forest ecosystems in ecotonal northern Alaska, USA, for the years 2003-2100. We compared energy feedbacks associated with increases in biomass to energy feedbacks associated with changes in the duration of the snow-free season. We based our simulations on nine input climate scenarios from the Intergovernmental Panel on Climate Change (IPCC) and a new version of the Terrestrial Ecosystem Model (TEM) that incorporates biogeochemistry, vegetation dynamics for multiple PFTs (e.g., trees, shrubs, grasses, sedges, mosses), multiple vegetation pools, and soil thermal regimes. We found mean increases in net primary productivity (NPP) in all PFTs. Most notably, birch (Betula spp.) in the shrub tundra showed increases that were at least three times larger than any other PFT. Increases in NPP were positively related to increases in growing-season length in the sedge tundra, but PFTs in boreal forest and shrub tundra showed a significant response to changes in light availability as well as growing-season length. Significant NPP responses to changes in vegetation uptake of nitrogen by PFT indicated that some PFTs were better competitors for nitrogen than other PFTs. While NPP increased, heterotrophic respiration (RH) also increased, resulting in decreases or no change in net ecosystem carbon uptake. Greater aboveground biomass from increased NPP produced a decrease in summer albedo, greater regional heat absorption (0.34 +/- 0.23 W x m(-2) x 10 yr(-1) [mean +/- SD]), and a positive feedback to climate warming. However, the decrease in albedo due to a shorter snow season (-5.1 +/- 1.6 d/10 yr) resulted in much greater regional heat absorption (3.3 +/- 1.24 W x m(-2) x 10 yr(-1)) than that associated with increases in vegetation. Through quantifying feedbacks associated with changes in vegetation and those associated with changes in the snow season length, we can reach a more integrated understanding of the manner in which climate change may impact interactions between high-latitude ecosystems and the climate system. PMID- 19544742 TI - Physiological responses to fertilization recorded in tree rings: isotopic lessons from a long-term fertilization trial. AB - Nitrogen fertilizer applications are common land use management tools, but details on physiological responses to these applications are often lacking, particularly for long-term responses over decades of forest management. We used tree ring growth patterns and stable isotopes to understand long-term physiological responses to fertilization using a controlled fertilization experiment begun in 1964 in Washington State (USA), in which three levels of nitrogen fertilizer were applied: 157, 314; and 471 kg/ha. Basal area increment (BAI) increased more than fourfold in the highest treatment to twofold in the lowest, and a significant increase in BAI was observed for 20 years. Latewood delta 13C sharply decreased by 1.4 per thousand after fertilization and was significantly lower than controls for four years, but no differences existed between fertilization levels, and the effect disappeared after four years, indicating that intrinsic water use efficiency (A/gs) increased in response to fertilization. Earlywood delta 13C showed similar trends but was more variable. Latewood delta 18O increased significantly above controls by approximately 2 per thousand in all treatments, but the duration differed with treatment level, with the effect being longer for higher levels of fertilization and lasting as long as nine years after fertilization. Because source water and relative humidity were the same between experimental plots, we interpreted the delta 18O increase with treatment as a decrease in leaf-level transpiration. Earlywood delta 18O did not show any treatment effects. Because the Pacific Northwest has a mediterranean climate with dry summers, we speculated that fertilization caused a substantial increase in leaf area, causing the trees to transpire themselves into drought stress during the late summer. We estimate from the delta 18O data that stomatal conductance (gs) was reduced by approximately 30%. Using the delta 13C data to estimate assimilation rates (A), A during the late season was also reduced by 20 30%. If leaf-level A decreased, but BAI increased, we estimated that leaf area on those trees must have increased by fourfold with the highest level of treatment within this stand. This increase in leaf area resulting from fertilization caused a hydraulic imbalance within the trees that lasted as long as nine years after treatment at the highest levels of fertilization. PMID- 19544743 TI - Legacies of historical land use on regional forest composition and structure in Wisconsin, USA (mid-1800s-1930s-2000s). AB - Historical land use can influence forest species composition and structure for centuries after direct use has ceased. In Wisconsin, USA, Euro-American settlement in the mid- to late 1800s was accompanied by widespread logging, agricultural conversion, and fire suppression. To determine the maximum magnitude of change in forest ecosystems at the height of the agricultural period and the degree of recovery since that time, we assessed changes in forest species composition and structure among the (1) mid-1800s, at the onset of Euro-American settlement; (2) 1930s, at the height of the agricultural period; and (3) 2000s, following forest regrowth. Data sources included the original U.S. Public Land Survey records (mid-1800s), the Wisconsin Land Economic Inventory (1930s), and U.S. Forest Service Forest Inventory and Analysis data (2000s). We derived maps of relative species dominance and tree diameters for the three dates and assessed change using spatial error models, nonmetric multidimensional scaling ordination, and Sorenson distance measures. Our results suggest that since the mid-1800s, hemlock and white pine have declined in absolute area from 22% to 1%, and the proportion of medium (25-<50 cm) and large-diameter (> or = 50 cm) trees of all species has decreased from 71% to 27% across the entire state. Early-successional aspen-birch is three times more common than in the mid-1800s (9% vs. 3%), and maple and other shade-tolerant species are increasing in southern areas formerly dominated by oak forests and savannas. Since the peak agricultural extent in the 1930s, species composition and tree size in northern forests have shown some recovery, while southern forests appear to be on a novel trajectory of change. There is evidence of regional homogenization, but the broad north-south environmental gradient in Wisconsin constrains overall species composition. Although the nature of the future forests will be determined in part by climate change and other exogenous variables, land use is likely to remain the driving factor. PMID- 19544744 TI - Sleepy synapses. Commentary on Hanlon et al. "Effects of skilled training on sleep slow wave activity and cortical gene expression in the rat" Sleep 2009;32:719-729. PMID- 19544745 TI - Natural history of sleep-disordered breathing: shedding light on the early years. Commentary on Bixler et al. Sleep disordered breathing in children in a general population sample: prevalence and risk factors. Sleep 2009;32(6):731-736. PMID- 19544746 TI - All work and no play makes Jack lose sleep. Commentary on Virtanen et al. Long working hours and sleep disturbances: the Whitehall II prospective cohort study. Sleep 2009;32:737-745. PMID- 19544747 TI - Effects of skilled training on sleep slow wave activity and cortical gene expression in the rat. AB - STUDY OBJECTIVE: The best characterized marker of sleep homeostasis is the amount of slow wave activity (SWA, 0.5-4 Hz) during NREM sleep. SWA increases as a function of previous waking time and declines during sleep, but the underlying mechanisms remain unclear. We have suggested that SWA homeostasis is linked to synaptic potentiation associated with learning during wakefulness. Indeed, studies in rodents and humans found that SWA increases after manipulations that presumably enhance synaptic strength, but the evidence remains indirect. Here we trained rats in skilled reaching, a task known to elicit long-term potentiation in the trained motor cortex, and immediately after learning measured SWA and cortical protein levels of c-fos and Arc, 2 activity-dependent genes involved in motor learning. DESIGN: Intracortical local field potential recordings and training on reaching task. SETTING: Basic sleep research laboratory. PATIENTS OR PARTICIPANTS: Long Evans adult male rats. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: SWA increased post-training in the trained cortex (the frontal cortex contralateral to the limb used to learn the task), with smaller or no increase in other cortical areas. This increase was reversible within 1 hour, specific to NREM sleep, and positively correlated with changes in performance during the prior training session, suggesting that it reflects plasticity and not just motor activity. Fos and Arc levels were higher in the trained relative to untrained motor cortex immediately after training, but this asymmetry was no longer present after 1 hour of sleep. CONCLUSION: Learning to reach specifically affects gene expression in the trained motor cortex and, in the same area, increases sleep need as measured by a local change in SWA. PMID- 19544749 TI - Long working hours and sleep disturbances: the Whitehall II prospective cohort study. AB - STUDY OBJECTIVE: To examine whether exposure to long working hours predicts various forms of sleep disturbance; short sleep, difficulty falling asleep, frequent waking, early waking and waking without feeling refreshed. DESIGN: Prospective study with 2 measurements of working hours (phase 3, 1991-1994 and phase 5, 1997-1999) and 2 measurements of subjective sleep disturbances (phase 5 and phase 7, 2002-2004). SETTING: The Whitehall II study of British civil servants. PARTICIPANTS: Full time workers free of sleep disturbances at phase 5 and employed at phases 5 and 7 (n = 937-1594) or at phases 3, 5, and 7 (n = 886 1510). MEASUREMENTS AND RESULTS: Working more than 55 hours a week, compared with working 35-40 hours a week, was related to incident sleep disturbances; demographics-adjusted odds ratio (95% CI) 1.98 (1.05, 3.76) for shortened sleeping hours, 3.68 (1.58, 8.58) for difficulty falling asleep; and 1.98 (1.04, 3.77) for waking without feeling refreshed. Repeat exposure to long working hours was associated with odds ratio 3.24 (1.45, 7.27) for shortened sleep, 6.66 (2.64, 16.83) for difficulty falling asleep, and 2.23 (1.16, 4.31) for early morning awakenings. Some associations were attenuated after adjustment for other risk factors. To a great extent, similar results were obtained using working hours as a continuous variable. Imputation of missing values supported the findings on shortened sleep and difficulty in falling asleep. CONCLUSION: Working long hours appears to be a risk factor for the development of shortened sleeping hours and difficulty falling asleep. PMID- 19544748 TI - Sleep disordered breathing in children in a general population sample: prevalence and risk factors. AB - STUDY OBJECTIVES: Assess the prevalence based on clinically meaningful criteria (i.e., blood pressure) and identify risk factors of sleep disordered breathing (SDB) in a representative sample of elementary school children. DESIGN: A random sample of the local elementary school children (K-5) were assessed using a two phased strategy. In phase I a brief questionnaire was completed by a parent of each child in local elementary schools (N = 5,740), with a response rate of 78.5%. In phase II, randomly selected children and their parent spent a night in our sleep laboratory (N = 700) with a response rate of 70.0%. SETTING: University sleep laboratory. PARTICIPANTS: Children enrolled in local elementary schools. INTERVENTION: None. MEASUREMENT & RESULTS: Each child was assessed with a full polysomnogram and completed a history/physical examination including an electrocardiogram, otolaryngology examination, and pulmonary evaluation. The prevalence of moderate SDB (apnea-hypopnea index > or = 5) was 1.2%. The independent risk factors included nasal abnormalities and minority associated only with mild (1 < AHI < 5) SDB and snoring and waist circumference associated with all levels of SDB. Tonsil size, based on visual inspection, was not an independent risk factor. CONCLUSION: The prevalence of AHI > or = 5 was 1.2% in a representative sample of elementary school children. Risk factors for SDB included waist circumference, nasal abnormalities (e.g., chronic sinusitis/rhinitis), and minority. The strong linear relationship between waist circumference and BMI across all degrees of severity of SDB suggests that, as in adults, metabolic factors may be among the most important risk factors for SDB in children. PMID- 19544750 TI - Dubious bargain: trading sleep for Leno and Letterman. AB - STUDY OBJECTIVES: Sleeping less than 7 hours daily impairs alertness and is associated with increased obesity, morbidity, and mortality; yet up to 40% of US adults do so. Population data indicate work time is the primary activity reciprocally related to sleep time in the United States. Reducing work time and its economic benefits to increase sleep time may not be feasible for most of the population. We sought to identify waking activities under discretionary control and adjacent to the sleep period that would be a more feasible source for increasing sleep time. DESIGN/PARTICIPANTS: American Time Use Survey data from 21,475 respondents aged > or = 15 years were pooled for the years 2003-2006 to explore activities in 2-hour periods prior to going to bed and past getting up on weekdays. INTERVENTIONS: N/A. RESULTS: Long workers (> or = 8 hours) terminated bed time an average of 0.68 h earlier than short workers (< 8 hours, P < 0.0001) and 1.31 h earlier than respondents not working on the interview day (P < 0.001), but time of going to bed did not differ among groups (22:37 vs. 22:42 vs. 22:37, respectively, P = 0.385). Watching television was the primary activity people engaged in before going to bed, accounting for 55.6 min (46.3%) of the 2-h pre bed period. In the morning, travel time and work time increased steadily toward the end of the post-awakening 2-h period, accounting for 14.8% and 12.3%, respectively. CONCLUSIONS: Watching television may be an important social Zeitgeber for the time of going to bed. Watching less television in the evening and postponing work start time in the morning appear to be the candidate behavioral changes for achieving additional sleep. While the timing of work may not be flexible, giving up some TV viewing in the evening should be possible to reduce chronic sleep debt and promote adequate sleep in those who need it. PMID- 19544751 TI - Idiopathic hypersomnia with and without long sleep time: a controlled series of 75 patients. AB - OBJECTIVE: To characterize the clinical, psychological, and sleep pattern of idiopathic hypersomnia with and without long sleep time, and provide normative values for 24-hour polysomnography. SETTING: University Hospital. DESIGN: Controlled, prospective cohort. PARTICIPANTS: 75 consecutive patients (aged 34 +/ 12 y) with idiopathic hypersomnia and 30 healthy matched controls. INTERVENTION: Patients and controls underwent during 48 hours a face-to-face interview, questionnaires, human leukocyte antigen genotype, a night polysomnography and multiple sleep latency test (MSLT), followed by 24-h ad libitum sleep monitoring. RESULTS: Hypersomniacs had more fatigue, higher anxiety and depression scores, and more frequent hypnagogic hallucinations (24%), sleep paralysis (28%), sleep drunkenness (36%), and unrefreshing naps (46%) than controls. They were more frequently evening types. DQB1*0602 genotype was similarly found in hypersomniacs (24.2%) and controls (19.2%). Hypersomniacs had more frequent slow wave sleep after 06:00 than controls. During 24-h polysomnography, the 95% confidence interval for total sleep time was 493-558 min in controls, versus 672-718 min in hypersomniacs. There were 40 hypersomniacs with and 35 hypersomniacs without long ( > 600 min) sleep time. The hypersomniacs with long sleep time were younger (29 +/- 10 vs 40 +/- 13 y, P = 0.0002), slimmer (body mass index: 26 +/- 5 vs 23 +/- 4 kg/m2; P = 0.005), and had lower Horne-Ostberg scores and higher sleep efficiencies than those without long sleep time. MSLT latencies were normal (> 8 min) in 71% hypersomniacs with long sleep time. CONCLUSIONS: Hypersomnia, especially with long sleep time, is frequently associated with evening chronotype and young age. It is inadequately diagnosed using MSLT. PMID- 19544753 TI - Actigraphy in the assessment of insomnia: a quantitative approach. AB - STUDY OBJECTIVE: The lack of quantitative criteria for identifying insomnia using actigraphy represents an unresolved limit for the use of actigraphy in a clinical setting. The current study was conducted to evaluate the most efficient actigraphic parameter in the assessment of insomnia and to suggest preliminary quantitative actigraphic criteria (QAC). PARTICIPANTS AND MEASUREMENTS: Performing a retrospective study we recovered 408 actigraphic records from 3 sleep measure databases: 2 regarding insomnia patients (n = 126) and one normal sleepers (n = 282). We compared the 2 samples analyzing the following actigraphic sleep parameters: time in bed (TIB), sleep onset latency (SOL), total sleep time (TST), wake after sleep onset (WASO), sleep efficiency percentage (SE%), number of awakenings longer than 5 minutes (NA > 5) and mean motor activity (MA). Moreover, a linear discriminant function (LDF) was developed to identify and combine the most useful actigraphic sleep parameters to separate insomnia patients from normal sleepers. Using Youden index we calculated the preliminary QAC for each actigraphic sleep parameter and for LDF. Receiver operator characteristic (ROC) curves for classifying the accuracy of QAC were performed. RESULTS: All sleep parameters recorded by actigraphy significantly differentiated the 2 groups, except TIB. An LDF analysis showed that the most useful combination of actigraphic sleep parameters to assess insomnia was TST, SOL, and NA > 5, which obtained the best ROC and the best balance between positive and negative predictive values compared to any single actigraphic parameter. CONCLUSION: Actigraphy provided a satisfactory objective measurement of sleep quality in insomnia patients. The combination of TST, SOL, and NA > 5 proved the best way to assess insomnia using actigraphy. Acknowledging that the lack of a technological standard and some methodological limitations prevent us generalizing our results, we recommend additional studies on larger populations using different actigraph models. PMID- 19544752 TI - Nighttime blood pressure in normotensive subjects with chronic insomnia: implications for cardiovascular risk. AB - OBJECTIVE: To assess as whether insomniacs have higher nighttime blood pressure (BP) and a blunted day-to-night BP reduction, recognized markers of increased risk of cardiovascular morbidity and mortality. DESIGN: Prospective case-control study. SETTING: University hospital-based sleep research laboratory. PARTICIPANTS: Thirteen normotensive subjects with chronic primary insomnia (9 women, 42 +/- 7 y) and 13 sex- and age-matched good sleepers. MEASUREMENTS AND RESULTS: Subjects underwent 2-week sleep diary and 3 sleep studies to provide subjective and objective sleep variables, and 24-h beat-to-beat BP recording to provide daytime, night-time and day-to-night BP changes ([nighttime daytime]/daytime)*100) (BP dipping). Spectral analysis of the electroencephalogram (EEG) was also performed during sleep of night 3 to assess EEG activity in the beta frequency (16-32 Hz), a measure of brain cortical activation. Nighttime SBP was higher (111 +/- 15 vs 102 +/- 12 mm Hg, P < 0.01) and day-to-night SBP dipping was lower (-8% +/- 6% vs -15% +/- 5%, P < 0.01) in insomniacs than good sleepers. Insomniacs also had higher activity in EEG beta frequency (P < 0.05). Higher nighttime SBP and smaller SBP dipping were independently associated with increased EEG beta activity (P < 0.05). CONCLUSIONS: Higher nighttime SBP and blunted day-to-night SBP dipping are present in normotensive subjects with chronic insomnia and are associated with a hyperactivity of the central nervous system during sleep. An altered BP profile in insomniacs could be one mechanism implicated in the link between insomnia and cardiovascular morbidity and mortality documented in epidemiological studies. PMID- 19544754 TI - Polysomnographic and health-related quality of life correlates of restless legs syndrome in the Sleep Heart Health Study. AB - STUDY OBJECTIVES: Sleep disturbance is the primary clinical morbidity of restless legs syndrome (RLS). To date, sleep disturbance in RLS has been measured in (1) clinical samples with polysomnography (PSG) or (2) population-based samples by self-report. The objective of this study was to analyze sleep by PSG in a population-based sample with symptoms of RLS. DESIGN: Cross-sectional observational study. SETTING: Community-based. PARTICIPANTS: 3433 older men and women. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: RLS was evaluated using an 8-item self-administered questionnaire based on NIH diagnostic criteria and required symptoms occurring > or = five times per month and associated with at least moderate distress. Health-related quality of life (HRQOL) was determined using the SF-36. Unattended, in-home PSG was performed. Data were assessed using general linear models with adjustment for demographic, health-related variables, and apnea-hypopnea index (AHI). Subjects with RLS had longer adjusted mean sleep latency (39.8 vs 26.4 min, P < 0.0001) and higher arousal index (20.1 vs 18.0, P = 0.0145) than those without RLS. Sleep latency increased progressively as the frequency of RLS symptoms increased from 5-15 days per month to 6-7 days per week. No differences in sleep stage percentages were observed between participants with and without RLS. Subjects with RLS also reported poorer HRQOL in all physical domains as well as in the Mental Health and Vitality domains. CONCLUSIONS: These novel PSG data from a nonclinical, community-based sample of individuals with RLS document sleep disturbance in the home even in individuals with intermittent symptoms. PMID- 19544756 TI - Obstructive sleep apnea: a risk factor for work disability. AB - STUDY OBJECTIVE: To examine obstructive sleep apnea (OSA) as a risk factor for work disability. PATIENTS AND SETTING: Consecutive patients referred to the University of California San Francisco Sleep Disorders Center with suspected OSA (n = 183). DESIGN: All patients underwent overnight polysomnography after completing a written survey which assessed work disability due to sleep problems, occupational characteristics and excessive daytime sleepiness (EDS) defined as an Epworth Sleepiness Scale score > 10. RESULTS: Among 150 currently employed patients, 83 had OSA on polysomnography (apnea-hypopnea index > or = 5). Compared with patients in whom both OSA and EDS were absent, patients with the combination of OSA and EDS were at higher risk of both recent work disability (adjusted odds ratio [OR], 13.7; 95% confidence interval [CI], 3.9-48) and longer-term work duty modification (OR, 3.6; CI, 1.1-12). When either OSA or EDS were absent, the strength of the association with work disability was less than when both OSA and EDS were present. When OSA was examined without respect to EDS, patients with OSA were at increased risk of recent work disability relative to patients without OSA (OR 2.6; 95% CI 1.2-5.8), but the association of OSA with longer-term work duty modification did not meet standard criteria for statistical significance (OR = 2.0, 95% CI 0.8-5.0). CONCLUSIONS: The combination of OSA and EDS contributes to work disability, and OSA by itself contributes to recent work disability. These findings should highlight to employers and clinicians the importance of OSA in the workplace to encourage patients to be screened for OSA, particularly in situations of decreased productivity associated with EDS. PMID- 19544755 TI - Sleep disorders and their association with laboratory pain sensitivity in temporomandibular joint disorder. AB - STUDY OBJECTIVES: We characterized sleep disorder rates in temporomandibular joint disorder (TMD) and evaluated possible associations between sleep disorders and laboratory measures of pain sensitivity. DESIGN: Research diagnostic examinations were conducted, followed by two consecutive overnight polysomnographic studies with morning and evening assessments of pain threshold. SETTING: Orofacial pain clinic and inpatient sleep research facility. PARTICIPANTS: Fifty-three patients meeting research diagnostic criteria for myofascial TMD. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: We determined sleep disorder diagnostic rates and conducted algometric measures of pressure pain threshold on the masseter and forearm. Heat pain threshold was measured on the forearm; 75% met self-report criteria for sleep bruxism, but only 17% met PSG criteria for active sleep bruxism. Two or more sleep disorders were diagnosed in 43% of patients. Insomnia disorder (36%) and sleep apnea (28.4%) demonstrated the highest frequencies. Primary insomnia (PI) (26%) comprised the largest subcategory of insomnia. Even after controlling for multiple potential confounds, PI was associated with reduced mechanical and thermal pain thresholds at all sites (P < 0.05). Conversely, the respiratory disturbance index was associated with increased mechanical pain thresholds on the forearm (P < 0.05). CONCLUSIONS: High rates of PI and sleep apnea highlight the need to refer TMD patients complaining of sleep disturbance for polysomnographic evaluation. The association of PI and hyperalgesia at a nonorofacial site suggests that PI may be linked with central sensitivity and could play an etiologic role in idiopathic pain disorders. The association between sleep disordered breathing and hypoalgesia requires further study and may provide novel insight into the complex interactions between sleep and pain-regulatory processes. PMID- 19544758 TI - Logging on for better sleep: RCT of the effectiveness of online treatment for insomnia. AB - STUDY OBJECTIVES: Despite effective cognitive behavioral treatments for chronic insomnia, such treatments are underutilized. This study evaluated the impact of a 5-week, online treatment for insomnia. DESIGN: This was a randomized controlled trial with online treatment and waiting list control conditions. PARTICIPANTS: Participants were 118 adults with chronic insomnia. SETTING: Participants received online treatment from their homes. INTERVENTION: Online treatment consisted of psychoeducation, sleep hygiene, and stimulus control instruction, sleep restriction treatment, relaxation training, cognitive therapy, and help with medication tapering. MEASUREMENT AND RESULTS: From pre- to post-treatment, there was a 33% attrition rate, and attrition was related to referral status (i.e., dropouts were more likely to have been referred for treatment rather than recruited from the community). Using a mixed model analysis of variance procedure (ANOVA), results showed that online treatment produced statistically significant improvements in the primary end points of sleep quality, insomnia severity, and daytime fatigue. Online treatment also produced significant changes in process variables of pre-sleep cognitive arousal and dysfunctional beliefs about sleep. CONCLUSIONS: Implications of these findings are that identification of who most benefits from online treatment is a worthy area of future study. PMID- 19544757 TI - Neighborhood of residence is associated with daily adherence to CPAP therapy. AB - STUDY OBJECTIVES: Adherence to continuous positive airway pressure (CPAP) therapy for obstructive sleep apnea is poor. Risk factors for nonadherence are not well understood but may reflect individual or neighborhood socioeconomic factors. We sought to determine the association of socioeconomic status and initial CPAP adherence. DESIGN: Retrospective cohort study, 2005 to 2006. SETTING: Philadelphia VA Medical Center. PARTICIPANTS: Of 330 consecutive veterans who met study criteria for initiation of CPAP therapy for newly diagnosed sleep apnea, 266 had complete data for study inclusion. INTERVENTIONS: N/A. MEASUREMENTS: Through a multivariable logistic regression model, using an outcome of objectively measured CPAP use - 4 h daily during the first week of treatment, we tested whether patients from higher socioeconomic neighborhoods had higher CPAP adherence. We measured neighborhood socioeconomic status with an index derived from the 2000 U.S. Census at the block group-level composed of median household income, male and female employment, adult high school completion, married households, and minority composition. RESULTS: CPAP adherence > 4 h occurred on 48.9% of 1,805 patient-days observed for the 266 subjects. After adjustment for individual sociodemographic characteristics and medical comorbidity, the probability of daily CPAP use 4 h ranged from 34.1% (95% CI, 26.4-42.7) for subjects from a low socioeconomic neighborhood (5th percentile) to 62.3% (95% CI, 53.8-70.1) for subjects from a high (95th percentile) neighborhood. CONCLUSIONS: In a retrospective cohort of veterans, initial CPAP adherence was closely associated with higher neighborhood socioeconomic factors. Future investigation should target specific impediments to adherence in the home and neighborhood environment. PMID- 19544760 TI - Point/counterpoint. Molecular breast imaging will soon replace x-ray mammography as the imaging modality of choice for women at high risk with dense breasts. PMID- 19544759 TI - Cost-effectiveness of eszopiclone for the treatment of adults with primary chronic insomnia. AB - STUDY OBJECTIVE: To assess the cost-effectiveness of treatment with eszopiclone for chronic primary insomnia in adults. METHODS: A model using patient-level data from a 6-month, double-blind, placebo-controlled, clinical trial (n = 824), combined with data from a claims database and published literature, was used to assess the quality-adjusted life years (QALYs) gained and costs associated with eszopiclone versus placebo in adults with primary insomnia. Quality of life data were collected during the trial via the SF-36, from which preference-based utility scores were derived using published algorithms. Medical and absenteeism costs, estimated via a retrospective analysis of a claims and absenteeism database, were assigned to patients based on the degree of severity of their insomnia, assessed via the Insomnia Severity Index collected in the clinical trial. Presenteeism costs (lost productivity while at work) were estimated from responses to the Work Limitation Questionnaire collected during the trial. Six month gains in QALYs and costs for each treatment group were calculated to derive cost-effectiveness ratios. Uncertainty was addressed via univariate and multivariate sensitivity analyses. RESULTS: Over the 6-month period, eszopiclone use resulted in a net gain of 0.0137 QALYs over placebo at an additional cost of $67, resulting in an incremental cost per QALY gained of slightly less than $5,000. When absenteeism and presenteeism costs were excluded, the cost effectiveness ratio increased to approximately $33,000 per QALY gained, which is below the commonly used threshold of $50,000 used to define cost-effectiveness. Extensive sensitivity analyses indicate the results are generally robust. CONCLUSION: Our model, based on efficacy data from a clinical trial, demonstrated eszopiclone was cost-effective for the treatment of primary insomnia in adults, especially when lost productivity costs were included. PMID- 19544761 TI - Design and evaluation of a methodology to perform personalized visual biofeedback for reducing respiratory amplitude in radiation treatment. AB - A methodology to perform personalized visual biofeedback aimed to the reduction of respiratory amplitude is here proposed. A custom-made software allows to adapt the biofeedback parameters to a patient's respiratory pattern by calculating a limiting range for respiratory amplitude obtained from data acquired during free breathing. The proposed methodology has been tested on ten healthy volunteers and on five lung cancer patients undergoing radiotherapy treatment. The protocol for volunteers consisted of 3 min of data acquisition during the subject's free breathing, 2 min of visual biofeedback within the limits, and 3 min of free breathing. The patients' free breathing was acquired in 3 min and the visual biofeedback performed during all the sessions of the radiotherapy treatment, i.e., an average of eight sessions and an average total treatment time of 2000 s each patient. All the volunteers and three patients of the five found the protocol comfortable. The settlement time needed for considering the limiting range stabilized during free breathing has been calculated as 120 +/- 10 s (p < 0.05). During visual biofeedback the baseline shift was removed and the average respiratory amplitude was reduced by about 40% for all the subjects. The variability of the breathing amplitude remained unaltered during biofeedback. Eight volunteers and three patients remained within the limiting range for more than 90% of the biofeedback period; all subjects remained within the limiting range for more than 80% of the biofeedback period. During the biofeedback period both groups, volunteers and patients, showed a significant increase in breathing frequency which was mostly doubled. Patients with shallow breathing performed comfortably the biofeedback. PMID- 19544762 TI - Ophthalmic applicators: an overview of calibrations following the change to SI units. AB - Since the NIST dose to water standard for 90Sr/90Y ophthalmic applicators was introduced, numerous sources have undergone calibration either at NIST or at the University of Wisconsin Accredited Dosimetry Calibration Laboratory (UWADCL). From 1997 to 2008, 222 of these beta-emitting sources were calibrated at the UWADCL, and prior reference source strength values were available for 149 of these sources. A survey of UWADCL ophthalmic applicator calibrations is presented here, demonstrating an average discrepancy of -19% with a standard deviation of +/- 16% between prior reference values and the NIST-traceable UWADCL absorbed dose to water calibrations. Values ranged from -49% to +42%. PMID- 19544764 TI - Operation logic and functionality of automatic dose rate and image quality control of conventional fluoroscopy. AB - New generation of fluoroscopic imaging systems is equipped with spectral shaping filters complemented with sophisticated automatic dose rate and image quality control logic called "fluoroscopy curve" or "trajectory." Such fluoroscopy curves were implemented first on cardiovascular angiographic imaging systems and are now available on conventional fluoroscopy equipment. This study aims to investigate the control logic operations under the fluoroscopy mode and acquisition mode (equivalent to the legacy spot filming) of a conventional fluoroscopy system typically installed for upper-lower gastrointestinal examinations, interventional endoscopy laboratories, gastrointestinal laboratory, and pain clinics. PMID- 19544763 TI - Liquid scintillator for 2D dosimetry for high-energy photon beams. AB - Complex radiation therapy techniques require dosimetric verification of treatment planning and delivery. The authors investigated a liquid scintillator (LS) system for application for real-time high-energy photon beam dosimetry. The system was comprised of a transparent acrylic tank filled with liquid scintillating material, an opaque outer tank, and a CCD camera. A series of images was acquired when the tank with liquid scintillator was irradiated with a 6 MV photon beam, and the light data measured with the CCD camera were filtered to correct for scattering of the optical light inside the liquid scintillator. Depth-dose and lateral profiles as well as two-dimensional (2D) dose distributions were found to agree with results from the treatment planning system. Further, the corrected light output was found to be linear with dose, dose rate independent, and is robust for single or multiple acquisitions. The short time needed for image acquisition and processing could make this system ideal for fast verification of the beam characteristics of the treatment machine. This new detector system shows a potential usefulness of the LS for 2D QA. PMID- 19544765 TI - Pediatric organ dose measurements in axial and helical multislice CT. AB - An anthropomorphic pediatric phantom (5-yr-old equivalent) was used to determine organ doses at specific surface and internal locations resulting from computed tomography (CT) scans. This phantom contains four different tissue-equivalent materials: Soft tissue, bone, brain, and lung. It was imaged on a 64-channel CT scanner with three head protocols (one contiguous axial scan and two helical scans [pitch = 0.516 and 0.984]) and four chest protocols (one contiguous axial scan and three helical scans [pitch = 0.516, 0.984, and 1.375]). Effective mA s [= (tube current x rotation time)/pitch] was kept nearly constant at 200 effective mA s for head and 290 effective mA s for chest protocols. Dose measurements were acquired using thermoluminescent dosimeter powder in capsules placed at locations internal to the phantom and on the phantom surface. The organs of interest were the brain, both eyes, thyroid, sternum, both breasts, and both lungs. The organ dose measurements from helical scans were lower than for contiguous axial scans by 0% to 25% even after adjusting for equivalent effective mA s. There was no significant difference (p > 0.05) in organ dose values between the 0.516 and 0.984 pitch values for both head and chest scans. The chest organ dose measurements obtained at a pitch of 1.375 were significantly higher than the dose values obtained at the other helical pitches used for chest scans (p < 0.05). This difference was attributed to the automatic selection of the large focal spot due to a higher tube current value. These findings suggest that there may be a previously unsuspected radiation dose benefit associated with the use of helical scan mode during computed tomography scanning. PMID- 19544766 TI - Patient-specific finite element modeling of respiratory lung motion using 4D CT image data. AB - Development and optimization of methods for adequately accounting for respiratory motion in radiation therapy of thoracic tumors require detailed knowledge of respiratory dynamics and its impact on corresponding dose distributions. Thus, computer aided modeling and simulation of respiratory motion have become increasingly important. In this article a biophysical approach for modeling respiratory lung motion is described: Major aspects of the process of lung ventilation are formulated as a contact problem of elasticity theory which is solved by finite element methods; lung tissue is assumed to be isotropic, homogeneous, and linearly elastic. A main focus of the article is to assess the impact of biomechanical parameters (values of elastic constants) on the modeling process and to evaluate modeling accuracy. Patient-specific models are generated based on 4D CT data of 12 lung tumor patients. Simulated motion patterns of inner lung landmarks are compared with corresponding motion patterns observed in the 4D CT data. Mean absolute differences between model-based predicted landmark motion and corresponding breathing-induced landmark displacements as observed in the CT data sets are in the order of 3 mm (end expiration to end inspiration) and 2 mm (end expiration to midrespiration). Modeling accuracy decreases with increasing tumor size both locally (landmarks close to tumor) and globally (landmarks in other parts of the lung). The impact of the values of the elastic constants appears to be small. Outcomes show that the modeling approach is an adequate strategy in predicting lung dynamics due to lung ventilation. Nevertheless, the decreased prediction quality in cases of large tumors demands further study of the influence of lung tumors on global and local lung elasticity properties. PMID- 19544767 TI - New empirical formula for neutron dose level at the maze entrance of 15 MV medical accelerator facilities. AB - An easily applicable empirical formula was derived for use in the assessment of the photoneutron dose at the maze entrance of a 15 MV medical accelerator treatment room. The neutron dose equivalent rates around the Varian medical accelerator head calculated with the Monte Carlo code MCNPX were used as the source term in producing the base data. The dose equivalents were validated by measurements with bubble detectors. Irradiation geometry conditions expected to yield higher neutron dose rates in the maze were selected: a 20 x 20 cm2 irradiation field, gantry rotation plane parallel to the maze walls, and the photon beams directed to the opposite wall to the maze entrance. The neutron dose equivalents at the maze entrance were computed for 697 arbitrary single-bend maze configurations by extending the Monte Carlo calculations down to the maze entrance. Then, the empirical formula was derived by a multiple regression fit to the neutron dose equivalents at the maze entrance for all the different maze configurations. The goodness of the empirical formula was evaluated by applying it to seven operating medical accelerators of different makes. When the source terms were fixed, the neutron doses estimated from the authors' formula agreed better with the corresponding MCNPX simulations than the results of the Kersey method. In addition, compared with the Wu-McGinley formula, the authors' formula provided better estimates for the mazes with length longer than 8.5 m. There are, however, discrepancies between the measured dose rates and the estimated values from the authors' formula, particularly for the machines other than a Varian model. Further efforts are needed to characterize the neutron field at the maze entrance to reduce the discrepancies. Furthermore, neutron source terms for the machines other than a Varian model should be simulated or measured and incorporated into the formula for accurate extended application to a variety of models. PMID- 19544768 TI - Stochastic noise characteristics in matrix inversion tomosynthesis (MITS). AB - Matrix inversion tomosynthesis (MITS) uses known imaging geometry and linear systems theory to deterministically separate in-plane detail from residual tomographic blur in a set of conventional tomosynthesis ("shift-and-add") planes. A previous investigation explored the effect of scan angle (ANG), number of projections (N), and number of reconstructed planes (NP) on the MITS impulse response and modulation transfer function characteristics, and concluded that ANG = 20 degrees, N = 71, and NP = 69 is the optimal MITS imaging technique for chest imaging on our prototype tomosynthesis system. This article examines the effect of ANG, N, and NP on the MITS exposure-normalized noise power spectra (ENNPS) and seeks to confirm that the imaging parameters selected previously by an analysis of the MITS impulse response also yield reasonable stochastic properties in MITS reconstructed planes. ENNPS curves were generated for experimentally acquired mean-subtracted projection images, conventional tomosynthesis planes, and MITS planes with varying combinations of the parameters ANG, N, and NP. Image data were collected using a prototype tomosynthesis system, with 11.4 cm acrylic placed near the image receptor to produce lung-equivalent beam hardening and scattered radiation. Ten identically acquired tomosynthesis data sets (realizations) were collected for each selected technique and used to generate ensemble mean images that were subtracted from individual image realizations prior to noise power spectra (NPS) estimation. NPS curves were normalized to account for differences in entrance exposure (as measured with an ion chamber), yielding estimates of the ENNPS for each technique. Results suggest that mid- and high-frequency noise in MITS planes is fairly equivalent in magnitude to noise in conventional tomosynthesis planes, but low-frequency noise is amplified in the most anterior and posterior reconstruction planes. Selecting the largest available number of projections (N = 71) does not incur any appreciable additive electronic noise penalty compared to using fewer projections for roughly equivalent cumulative exposure. Stochastic noise is minimized by maximizing N and NP but increases with increasing ANG. The noise trend results for NP and ANG are contrary to what would be predicted by simply considering the MITS matrix conditioning and likely result from the interplay between noise correlation and the polarity of the MITS filters. From this study, the authors conclude that the previously determined optimal MITS imaging strategy based on impulse response considerations produces somewhat suboptimal stochastic noise characteristics, but is probably still the best technique for MITS imaging of the chest. PMID- 19544769 TI - A novel three-dimensional image reconstruction method for near-field coded aperture single photon emission computerized tomography. AB - Coded aperture imaging for two-dimensional (2D) planar objects has been investigated extensively in the past, whereas little success has been achieved in imaging 3D objects using this technique. In this article, the authors present a novel method of 3D single photon emission computerized tomography (SPECT) reconstruction for near-field coded aperture imaging. Multiangular coded aperture projections are acquired and a stack of 2D images is reconstructed separately from each of the projections. Secondary projections are subsequently generated from the reconstructed image stacks based on the geometry of parallel-hole collimation and the variable magnification of near-field coded aperture imaging. Sinograms of cross-sectional slices of 3D objects are assembled from the secondary projections, and the ordered subset expectation and maximization algorithm is employed to reconstruct the cross-sectional image slices from the sinograms. Experiments were conducted using a customized capillary tube phantom and a micro hot rod phantom. Imaged at approximately 50 cm from the detector, hot rods in the phantom with diameters as small as 2.4 mm could be discerned in the reconstructed SPECT images. These results have demonstrated the feasibility of the authors' 3D coded aperture image reconstruction algorithm for SPECT, representing an important step in their effort to develop a high sensitivity and high resolution SPECT imaging system. PMID- 19544770 TI - Differences among Monte Carlo codes in the calculations of voxel S values for radionuclide targeted therapy and analysis of their impact on absorbed dose evaluations. AB - Several updated Monte Carlo (MC) codes are available to perform calculations of voxel S values for radionuclide targeted therapy. The aim of this work is to analyze the differences in the calculations obtained by different MC codes and their impact on absorbed dose evaluations performed by voxel dosimetry. Voxel S values for monoenergetic sources (electrons and photons) and different radionuclides (90Y, 131I, and 188Re) were calculated. Simulations were performed in soft tissue. Three general-purpose MC codes were employed for simulating radiation transport: MCNP4C, EGSnrc, and GEANT4. The data published by the MIRD Committee in Pamphlet No. 17, obtained with the EGS4 MC code, were also included in the comparisons. The impact of the differences (in terms of voxel S values) among the MC codes was also studied by convolution calculations of the absorbed dose in a volume of interest. For uniform activity distribution of a given radionuclide, dose calculations were performed on spherical and elliptical volumes, varying the mass from 1 to 500 g. For simulations with monochromatic sources, differences for self-irradiation voxel S values were mostly confined within 10% for both photons and electrons, but with electron energy less than 500 keV, the voxel S values referred to the first neighbor voxels showed large differences (up to 130%, with respect to EGSnrc) among the updated MC codes. For radionuclide simulations, noticeable differences arose in voxel S values, especially in the bremsstrahlung tails, or when a high contribution from electrons with energy of less than 500 keV is involved. In particular, for 90Y the updated codes showed a remarkable divergence in the bremsstrahlung region (up to about 90% in terms of voxel S values) with respect to the EGS4 code. Further, variations were observed up to about 30%, for small source-target voxel distances, when low-energy electrons cover an important part of the emission spectrum of the radionuclide (in our case, for 131I). For 90Y and 188Re, the differences among the various codes have a negligible impact (within few percents) on convolution calculations of the absorbed dose; thus either one of the MC programs is suitable to produce voxel S values for radionuclide targeted therapy dosimetry. However, if a low-energy beta-emitting radionuclide is considered, these differences can affect also dose depositions at small source target voxel distances, leading to more conspicuous variations (about 9% for 1311) when calculating the absorbed dose in the volume of interest. PMID- 19544771 TI - A new automated method for the segmentation and characterization of breast masses on ultrasound images. AB - Segmentation is one of the first steps in most computer-aided diagnosis systems for characterization of masses as malignant or benign. In this study, the authors designed an automated method for segmentation of breast masses on ultrasound (US) images. The method automatically estimated an initial contour based on a manually identified point approximately at the mass center. A two-stage active contour method iteratively refined the initial contour and performed self-examination and correction on the segmentation result. To evaluate the method, the authors compared it with manual segmentation by two experienced radiologists (R1 and R2) on a data set of 488 US images from 250 biopsy-proven masses (100 malignant and 150 benign). Two area overlap ratios (AOR1 and AOR2) and an area error measure were used as performance measures to evaluate the segmentation accuracy. Values for AOR1, defined as the ratio of the intersection of the computer and the reference segmented areas to the reference segmented area, were 0.82 +/- 0.16 and 0.84 +/- 0.18, respectively, when manually segmented mass regions by R1 and R2 were used as the reference. Although this indicated a high agreement between the computer and manual segmentations, the two radiologists' manual segmentation results were significantly (p < 0.03) more consistent, with AOR1 = 0.84 +/- 0.16 and 0.91 +/- 0.12, respectively, when the segmented regions by R1 and R2 were used as the reference. To evaluate the segmentation method in terms of lesion classification accuracy, feature spaces were formed by extracting texture, width to-height, and posterior shadowing features based on either automated computer segmentation or the radiologists' manual segmentation. A linear discriminant analysis classifier was designed using stepwise feature selection and two-fold cross validation to characterize the mass as malignant or benign. For features extracted from computer segmentation, the case-based test A(z) values ranged from 0.88 +/- 0.03 to 0.92 +/- 0.02, indicating a comparable performance to those extracted from manual segmentation by radiologists (A(z) value range: 0.87 +/- 0.03 to 0.90 +/- 0.03). PMID- 19544773 TI - Algorithm and simulation for real-time positron emission based tumor tracking using a linear fiducial marker. AB - The effectiveness of radiotherapy in cancer treatment remains significantly limited by the accuracy of tumor dose delivery. The ideal solution lies in real time localization of patient tumors during therapy; one such method is by tracking implanted low-activity positron emitters using two pairs of orthogonally placed gamma-ray detectors. Prior studies have examined multiple point sources, which have potential patient complications during implantation. A linear source geometry is proposed as a less invasive alternative, with potential higher precision tracking. A source localization algorithm has been devised using cost function minimization of the source position estimate relative to annihilation gamma coincidence lines. The algorithm was tested via Monte Carlo simulation methods using a Geant4 application for emission tomography (GATE) package for a source of length of 2.00 cm and width of 0.1 mm. The midpoint of the linear marker was located within submillimeter accuracy at 200 coincidence events and the orientation of the source determined with less than 5 degrees (0.087 rad) angular deviation at 300 events. At an optimal event count of 700, tracking had mean midpoint error of 0.48 +/- 0.26 mm and mean angular deviation of 0.041 +/- 0.023 rad (1.4 degrees +/- 0.8 degree). The source and tracking algorithm may prove effective for future clinical implementation in radiotherapy treatment. PMID- 19544772 TI - Monte Carlo evaluation of the convolution/superposition algorithm of Hi-Art tomotherapy in heterogeneous phantoms and clinical cases. AB - The reliability of the convolution/superposition (C/S) algorithm of the Hi-Art tomotherapy system is evaluated by using the Monte Carlo model TomoPen, which has been already validated for homogeneous phantoms. The study was performed in three stages. First, measurements with EBT Gafchromic film for a 1.25 x 2.5 cm2 field in a heterogeneous phantom consisting of two slabs of polystyrene separated with Styrofoam were compared to simulation results from TomoPen. The excellent agreement found in this comparison justifies the use of TomoPen as the reference for the remaining parts of this work. Second, to allow analysis and interpretation of the results in clinical cases, dose distributions calculated with TomoPen and C/S were compared for a similar phantom geometry, with multiple slabs of various densities. Even in conditions of lack of lateral electronic equilibrium, overall good agreement was obtained between C/S and TomoPen results, with deviations within 3%/2 mm, showing that the C/S algorithm accounts for modifications in secondary electron transport due to the presence of a low density medium. Finally, calculations were performed with TomoPen and C/S of dose distributions in various clinical cases, from large bilateral head and neck tumors to small lung tumors with diameter of < 3 cm. To ensure a "fair" comparison, identical dose calculation grid and dose-volume histogram calculator were used. Very good agreement was obtained for most of the cases, with no significant differences between the DVHs obtained from both calculations. However, deviations of up to 4% for the dose received by 95% of the target volume were found for the small lung tumors. Therefore, the approximations in the C/S algorithm slightly influence the accuracy in small lung tumors even though the C/S algorithm of the tomotherapy system shows very good overall behavior. PMID- 19544774 TI - Solid water as phantom material for dosimetry of electron backscatter using low energy electron beams: a Monte Carlo evaluation. AB - This study evaluated the dosimetry of electron backscatter when Solid Water is used to substitute water as phantom in electron radiotherapy. Monte Carlo simulation (EGSnrc-based code) was employed to predict electron energy spectra and depth doses for the 0.5 and 1 cm of Solid Water and water slabs above 3 mm of lead (Pb) layers using electron beams with energies of 4 and 6 MeV. For comparison, Monte Carlo simulations were repeated with Pb layers taken out from the phantoms using the same experimental configuration. Analyses on electron energy spectra for the 4 and 6 MeV electron beams showed that deviations of electron energy distributions between the Solid Water and water phantom were more significant in the high-energy range (i.e., close to the maximal electron energy) than the lower range corresponding to the electron backscatter. These deviations of electron energy spectra varied with depth and were mainly due to the electron fluence or beam attenuation. Dosimetry results from Monte Carlo simulations showed that the Solid Water phantom had lower depth dose compared to water with the same experimental setup. For the 4 MeV electron beams with 0.5 cm of Solid Water, depth doses were 1.8%-3.9% and 2.3%-4.4% lower than those in water, with and without the Pb layer underneath, respectively. Thicker Solid Water of 1 cm resulted in different decreases in depth doses of 1.8%-4.6% (with Pb) and 2.3% 4.4% (without Pb) compared to water. For higher nominal electron beam energy of 6 MeV with 0.5 cm of Solid Water, depth doses decreased 1.7%-2.9% (with Pb) and 1.6%-2.1% (without Pb) compared to water. These decreases in depth doses changed to 1.7%-3.7% (with Pb) and 1.7%-3% (without Pb) when the thickness of Solid Water was increased to 1 cm. The dosimetry data in this study are useful in determining the correction factor when using Solid Water to substitute water for the electron backscatter measurement in electron radiotherapy. PMID- 19544775 TI - Comparison of organ residence time estimation methods for radioimmunotherapy dosimetry and treatment planning--patient studies. AB - The estimation of organ residence time is essential for high-dose myeloablative regimens in radioimmunotherapy (RIT). Frequently, this estimation is based on a series of simple planar scans and planar processing. The authors previously performed a simulation study which demonstrated that the accuracy of this methodology is limited compared to a hybrid planar/SPECT residence time estimation method. In this work the authors applied this hybrid method to data from a clinical trial of high-dose myeloablative yttrium-90 ibritumomab tiuxetan therapy. Image data acquired from 18 patients were comprised of planar scans at five time points ranging from 1 to 144 h postinjection and abdominal and thoracic SPECT/CT scans obtained at 24 h postinjection. The simple planar processing method used in this work was based on the geometric mean method with energy window based scatter compensation. No explicit background subtraction nor object or source thickness corrections were performed. The SPECT projections were reconstructed using iterative reconstruction with compensations for attenuation, scatter, and full collimator-detector response. Large differences were observed when residence times were estimated using the simple planar method compared to the hybrid method. The differences were not constant but varied in magnitude and sign. For the dose-limiting organ (liver), the average difference was -18% and variation in the difference was 19%, similar to the differences observed in a previously reported simulation study. The authors also looked at the relationship between the weight of the patient and the liver residence time and found that there was no meaningful correlation for either method. This indicates that weight would not be an adequate proxy for an experimental estimate of residence time when choosing the activity to administer for therapy. The authors conclude that methods such as the simple planar method used here are inadequate for RIT treatment planning. More sophisticated methods, such as the hybrid SPECT/planar method investigated here, are likely to be better predictors of organ dose and, as a result, organ toxicities. PMID- 19544777 TI - Impact of motion velocity on four-dimensional target volumes: a phantom study. AB - This study aims to assess the impact of motion velocity that may cause motion artifacts on target volumes (TVs) using a one-dimensional moving phantom. A 20 mm diameter spherical object embedded in a QUASAR phantom sinusoidally moved with approximately 5.0 or 10.0 mm amplitude (A) along the longitudinal axis of the computed tomography (CT) couch. The motion period was manually set in the range of 2.0-10.0 s at approximately 2.0 s interval. Four-dimensional (4D) CT images were acquired by a four-slice CT scanner (LightSpeed RT; General Electric Medical Systems, Waukesha, WI) with a slice thickness of 1.25 mm in axial cine mode. The minimum gantry rotation of 1.0 s was employed to achieve the maximum in-slice temporal resolution. Projection data over a full gantry rotation (1.0 s) were used for image reconstruction. Reflective marker position was recorded by the real-time positioning management system (Varian Medical Systems, Palo Alto, CA). ADVANTAGE 4D software exported ten respiratory phase volumes and the maximum intensity volume generated from all reconstructed data (MIV). The threshold to obtain static object volume (V0, 4.19 ml) was used to automatically segment TVs on CT images, and then the union of TVs on 4D CT images (TV(4D)) was constructed. TVs on MIV (TV(MIV)) were also segmented by the threshold that can determine the area occupied within the central slice of TV(MIV). The maximum motion velocity for each phase bin was calculated using the actual averaged motion period displayed on ADVANTAGE 4D software (T), the range of phases used to construct the target phase bin (phase range), and a mathematical model of sinusoidal function. Each volume size and the motion range of TV in the cranial-caudal (CC) direction were measured. Subsequently, cross-correlation coefficients between TV size and motion velocity as well as phase range were calculated. Both misalignment and motion-blurring artifacts were caused by high motion velocity, Less than 6% phase range was needed to construct the 4D CT data set, except for T of 2.0 s. While the positional differences between the TV and ideal centroid in the CC direction were within the voxel size for T > or = 6.0 s, the differences were up to 2.43 and 4.15 mm for (A,T) = (5.0 mm, 2.0 s) and (10.0 mm, 2.0 s), respectively. The maximum volumetric deviations between TV sizes and V0 were 43.68% and 91.41% for A of 5.0 and 10.0 mm, respectively. TV(MIV) sizes were slightly larger than TV(4D) sizes. Volumetric deviation between TV size and V0 had a stronger correlation with motion velocity rather than phase range. This phantom study demonstrated that motion artifacts were substantially reduced when the phantom moved longitudinally at low motion velocity during 4D CT image acquisition; therefore, geometrical uncertainties due to motion artifacts should be recognized when determining TVs, especially with a fast period. PMID- 19544776 TI - Quantitative imaging of element composition and mass fraction using dual-energy CT: three-material decomposition. AB - In principle, dual-energy CT can only accurately decompose a mixture into two materials. To decompose a mixture into three constitute materials using dual energy CT measurements, a third criteria must be provided to solve for three unknowns with only two spectral measurements. One solution is to assume that the sum of the volumes of three constituent materials is equivalent to the volume of the mixture (i.e., volume conservation), but this is not always true. A more generalized solution is to use the principle of mass conservation, which assumes that the sum of the masses of the three constituent materials is equivalent to the mass of the mixture. In this article, a mass-conservation based, three material decomposition dual-energy CT algorithm is described and experimental validation of the accuracy of the technique presented. The results demonstrate that the proposed method can accurately measure elemental concentrations under low noise imaging conditions. Clinically, this may be applied to measure the mass fraction of any chemical element in a three-material mixture of solutions without the requirement of volume conservation. PMID- 19544778 TI - Comparison of low-pitch and respiratory-averaged CT protocols for attenuation correction of cardiac PET studies. AB - PET/CT perfusion studies suffer from artifacts caused by misalignment of transmission and emission data due to contractile cardiac and respiratory motion. This study investigates whether substantial differences exist between two respiration-averaging approaches for attenuation correction (AC): low-pitch helical (HCT) and time-averaged CT (ACT). Fifty-four consecutive patients received paired HCT (0.45 pitch, 120 kVp, 76 mA, 24 x 1.2 mm collimated slice width, 1 s gantry rotation time, 4.93 mGy CTDI) and ACT (sequence mode: 6.1 s acq/bed, 80 kVp, 13 mA, 24 x 1.2 mm collimated slice width, 5.53 mGy CTDI) AC scans under free-breathing prior to Rb-82 rest/adenosine stress. Mismatch between the emission and paired transmission data was compared by calculating the volume of myocardial uptake overlying the left CT lung field. Data were then reconstructed with the CT AC scans and normalized to injected dose and bodyweight. Paired rest and paired stress PET images were reoriented identically along the short axis and sampled into a 17-segment polar map for comparison. The ratio of HCT-PET and ACT-PET polar maps at rest and stress was calculated and grouped by segment for all patients. 95% confidence intervals were calculated to compare changes in the polar map ratios between the two AC methods. No significant difference was observed between the HCT and ACT overlying volume in the rest or stress emission data. 68% of the patients presented visual respiratory artifacts in the HCT images compared to 32% in the ACT. That 23% of the ACT images presented with photon starvation artifacts and increasing BMI was a significant indicator for the occurrence of photon starvation in the ACT AC scans (p < 0.001). The ratio of the reconstructed PET polar segment data showed good agreement between AC methods with 95% confidence intervals ranging from 0.92 to 1.07 in the rest data and 0.93 to 1.07 in the stress data segments. Bias, calculated by averaging the polar segment ratios, showed 1% higher values in the ACT-PET rest reconstructions compared to the HCT-PET rest reconstructions and no measurable bias in the stress reconstructions. This study shows good agreement and negligible bias between low-pitch HCT and ACT protocols for attenuation correction of cardiac PET data. PMID- 19544779 TI - Fixed gantry tomosynthesis system for radiation therapy image guidance based on a multiple source x-ray tube with carbon nanotube cathodes. AB - The authors present the design and simulation of an imaging system that employs a compact multiple source x-ray tube to produce a tomosynthesis image from a set of projections obtained at a single tube position. The electron sources within the tube are realized using cold cathode carbon nanotube technology. The primary intended application is tomosynthesis-based 3D image guidance during external beam radiation therapy. The tube, which is attached to the gantry of a medical linear accelerator (linac) immediately below the multileaf collimator, operates within the voltage range of 80-160 kVp and contains a total of 52 sources that are arranged in a rectilinear array. This configuration allows for the acquisition of tomographic projections from multiple angles without any need to rotate the linac gantry. The x-ray images are captured by the same amorphous silicon flat panel detector employed for portal imaging on contemporary linacs. The field of view (FOV) of the system corresponds to that part of the volume that is sampled by rays from all sources. The present tube and detector configuration provides an 8 x 8 cm2 FOV in the plane of the linac isocenter when the 40.96 x 40.96 cm2 imaging detector is placed 40 cm from the isocenter. Since this tomosynthesis application utilizes the extremities of the detector to record image detail relating to structures near the isocenter, simultaneous treatment and imaging is possible for most clinical cases, where the treated target is a small region close to the linac isocenter. The tomosynthesis images are reconstructed using the simultaneous iterative reconstruction technique, which is accelerated using a graphic processing unit. The authors present details of the system design as well as simulated performance of the imaging system based on reprojections of patient CT images. PMID- 19544780 TI - Assessment of the limiting spatial resolution of an MRI scanner by direct analysis of the edge spread function. AB - Limiting spatial resolution is a key metric of the quality of magnetic resonance (MR) images, which can provide an indication of the smallest region that can effectively be imaged. In this paper a novel methodology for measuring the limiting spatial resolution of MR images is mathematically analyzed and successfully implemented on phantom images. The methodology presented in this paper is based on a direct fit of a mathematical expression of the edge spread function (ESF) profile to the ESF data acquired at the interface between different materials. The mathematical expression of ESF was determined by approximating the line spread function (LSF) of the system with a sinc function. The proposed methodology can be applied using signal data from magnitude MRI spin echo images and is not sensitive to noise amplification introduced by differentiating the ESF to produce the LSF, as performed in previous studies. In addition, the proposed methodology provides a quantitative, representative measurement of the limiting spatial resolution of MR images. PMID- 19544783 TI - Sinogram bow-tie filtering in FBP PET reconstruction. AB - Low-pass filtering of sinograms in the radial direction is the most common practice to limit noise amplification in filtered back projection (FBP) reconstruction of positron emission tomography studies. Other filtering strategies have been proposed to prevent the loss in resolution due to low-pass radial filters, although results have been diverse. Using the well-known properties of the Fourier transform of a sinogram, the authors defined a binary mask that matches the expected shape of the support region in the Fourier domain of the sinogram ("bow tie"). This mask was smoothed by a convolution with a ten point Gaussian kernel which not only avoids ringing but also introduces a pre emphasis at low frequencies. A new filtering scheme for FBP is proposed, comprising this smoothed bow-tie filter combined with a standard radial filter and an axial filter. The authors compared the performance of the bow-tie filtering scheme with that of other previously reported methods: Standard radial filtering, angular filtering, and stackgram-domain filtering. All the quantitative data in the comparisons refer to a baseline reconstruction using a ramp filter only. When using the smallest size of the Gaussian kernel in the stackgram domain, the authors achieved a noise reduction of 33% at the cost of degrading radial and tangential resolutions (14.5% and 16%, respectively, for cubic interpolation). To reduce the noise by 30%, the angular filter produced a larger degradation of contrast (3%) and tangential resolution (46% at 10 mm from the center of the field of view) and showed noticeable artifacts in the form of circular blurring dependent on the distance to the center of the field of view. For a similar noise reduction (33%), the proposed bow-tie filtering scheme yielded optimum results in resolution (gain in radial resolution of 10%) and contrast (1% increase) when compared with any of the other filters alone. Experiments with rodent images showed noticeable image quality enhancement when using the proposed bow-tie filtering scheme. PMID- 19544782 TI - Comparison of online IGRT techniques for prostate IMRT treatment: adaptive vs repositioning correction. AB - This study compares three online image guidance techniques (IGRT) for prostate IMRT treatment: bony-anatomy matching, soft-tissue matching, and online replanning. Six prostate IMRT patients were studied. Five daily CBCT scans from the first week were acquired for each patient to provide representative "snapshots" of anatomical variations during the course of treatment. Initial IMRT plans were designed for each patient with seven coplanar 15 MV beams on a Eclipse treatment planning system. Two plans were created, one with a PTV margin of 10 mm and another with a 5 mm PTV margin. Based on these plans, the delivered dose distributions to each CBCT anatomy was evaluated to compare bony-anatomy matching, soft-tissue matching, and online replanning. Matching based on bony anatomy was evaluated using the 10 mm PTV margin ("bone10"). Soft-tissue matching was evaluated using both the 10 mm ("soft10") and 5 mm ("soft5") PTV margins. Online reoptimization was evaluated using the 5 mm PTV margin ("adapt"). The replanning process utilized the original dose distribution as the basis and linear goal programming techniques for reoptimization. The reoptimized plans were finished in less than 2 min for all cases. Using each IGRT technique, the delivered dose distribution was evaluated on all 30 CBCT scans (6 patients x 5 CBCT/patient). The mean minimum dose (in percentage of prescription dose) to the CTV over five treatment fractions were in the ranges of 99%-100% (SD = 0.1% 0.8%), 65%-98% (SD = 0.4%-19.5%), 87%-99% (SD = 0.7%-23.3%), and 95%-99% (SD = 0.4%-10.4%) for the adapt, bone10, soft5, and soft10 techniques, respectively. Compared to patient position correction techniques, the online reoptimization technique also showed improvement in OAR sparing when organ motion/deformations were large. For bladder, the adapt technique had the best (minimum) D90, D50, and D30 values for 24, 17, and 15 fractions out of 30 total fractions, while it also had the best D90, D50, and D30 values for the rectum for 25, 16, and 19 fractions, respectively. For cases where the adapt plans did not score the best for OAR sparing, the gains of the OAR sparing in the repositioning-based plans were accompanied by an underdosage in the target volume. To further evaluate the fast online replanning technique, a gold-standard plan ("new" plan) was generated for each CBCT anatomy on the Eclipse treatment planning system. The OAR sparing from the online replanning technique was compared to the new plan. The differences in D90, D50, and D30 of the OARs between the adapt and the new plans were less than 5% in 3 patients and were between 5% and 10% for the remaining three. In summary, all IGRT techniques could be sufficient to correct simple geometrical variations. However, when a high degree of deformation or differential organ position displacement occurs, the online reoptimization technique is feasible with less than 2 min optimization time and provides improvements in both CTV coverage and OAR sparing over the position correction techniques. For these cases, the reoptimization technique can be a highly valuable online IGRT tool to correct daily treatment uncertainties, especially when hypofractionation scheme is applied and daily correction, rather than averaging over many fractions, is required to match the original plan. PMID- 19544781 TI - Implementation and validation of an ultrasonic tissue characterization technique for quantitative assessment of normal-tissue toxicity in radiation therapy. AB - The goal of this study was to implement and validate a noninvasive, quantitative ultrasonic technique for accurate and reproducible measurement of normal-tissue toxicity in radiation therapy. The authors adapted an existing ultrasonic tissue characterization (UTC) technique that used a calibrated 1D spectrum based on region-of-interest analysis. They modified the calibration procedure by using a reference phantom instead of a planar reflector. This UTC method utilized ultrasonic radiofrequency echo signals to generate spectral parameters related to the physical properties (e.g., size, shape, and relative acoustic impedance) of tissue microstructures. Three spectral parameters were investigated for quantification of normal-tissue injury: Spectral slope, intercept, and midband fit. They conducted a tissue-mimicking phantom study to verify the reproducibility of UTC measurements and initiated a clinical study of radiation induced breast-tissue toxicity. Spectral parameter values from measurements on two phantoms were reproducible within 1% of each other. Eleven postradiation breast-cancer patients were studied and significant differences between the irradiated and untreated (contralateral) breasts were observed for spectral intercept (p = 0.003) and midband fit (p < 0.001) but not for slope (p = 0.14). In comparison to the untreated breast, the average difference in the spectral intercept was 2.99 +/- 0.75 dB and the average difference in the midband fit was 3.99 +/- 0.65 dB. The preliminary clinical study demonstrated the feasibility of using the quantitative ultrasonic method to evaluate normal-tissue toxicity in radiation therapy. PMID- 19544784 TI - Evaluation of a novel 4D in vivo dosimetry system. AB - A prototype of a new 4D in vivo dosimetry system capable of simultaneous real time position monitoring and dose measurement has been developed. The radiation positioning system (RADPOS) is controlled by a computer and combines two technologies: MOSFET radiation detector coupled with an electromagnetic positioning device. Special software has been developed that allows sampling position and dose either manually or automatically in user-defined time intervals. Preliminary tests of the new device include a dosimetric evaluation of the detector in 60Co, 6 MV, and 18 MV beams and measurements of spatial position stability and accuracy. In addition, the effect of metals and other materials on the performance of the positioning system has been investigated. Results show that the RADPOS system can measure in-air dose profiles that agree, on average, within 3%-5% of diode measurements for the energies tested. The response of the detector is isotropic within 1.6% (1 SD) with a maximum deviation of +/- 4.0% over 360 degrees. The maximum variation in the calibration coefficient over field sizes from 6 x 6 to 25 x 25 cm2 was 2.3% for RADPOS probe with the high sensitivity MOSFET and 4.6% for the probe with the standard sensitivity MOSFET. Of the materials tested, only aluminum, lead, and brass caused shifts in the RADPOS read position. The magnitude of the shift varied between materials and size of the material sample. Nonmagnetic stainless steel (Grade 304) caused a distortion of less than 2 mm when placed within 10 mm of the detector; therefore, it can provide a reasonable alternative to other metals if required. The results of the preliminary tests indicate that the device can be used for in vivo dosimetry in 60Co and high-energy beams from linear accelerators. PMID- 19544786 TI - The energy dependence and dose response of a commercial optically stimulated luminescent detector for kilovoltage photon, megavoltage photon, and electron, proton, and carbon beams. AB - Optically stimulated luminescent detectors, which are widely used in radiation protection, offer a number of potential advantages for application in radiation therapy dosimetry. Their introduction into this field has been somewhat hampered by the lack of information on their radiation response in megavoltage beams. Here the response of a commercially available optically stimulated luminescent detector (OSLD) is determined as a function of energy, absorbed dose to water, and linear energy transfer (LET). The detector response was measured as a function of energy for absorbed doses from 0.5 to 4.0 Gy over the following ranges: 125 kVp to 18 MV for photons, 6-20 MeV for electrons, 50-250 MeV for protons, and 290 MeV/u for the carbon ions. For the low LET beams, the response of the detector was linear up to 2 Gy with supralinearity occurring at higher absorbed doses. For the kilovoltage photons, the detector response relative to 6 MV increased with decreasing energy due to the higher atomic number of aluminum oxide (11.2) relative to water (7.4). For the megavoltage photons and electrons, the response was independent of energy. The response for protons was also independent of energy, but it was about 6% higher than its response to 6 MV photons. For the carbon ions, the dose response was linear for a given LET from 0.5 to 4.0 Gy, and no supralinearity was observed. However, it did exhibit LET dependence on the response relative to 6 MV photons decreasing from 1.02 at 1.3 keV/microm to 0.41 at 78 keV/microm. These results provide additional information on the dosimetric properties for this particular OSL detector and also demonstrate the potential for their use in photon, electron, and proton radiotherapy dosimetry with a more limited use in high LET radiotherapy dosimetry. PMID- 19544785 TI - Analysis of outcomes in radiation oncology: an integrated computational platform. AB - Radiotherapy research and outcome analyses are essential for evaluating new methods of radiation delivery and for assessing the benefits of a given technology on locoregional control and overall survival. In this article, a computational platform is presented to facilitate radiotherapy research and outcome studies in radiation oncology. This computational platform consists of (1) an infrastructural database that stores patient diagnosis, IMRT treatment details, and follow-up information, (2) an interface tool that is used to import and export IMRT plans in DICOM RT and AAPM/RTOG formats from a wide range of planning systems to facilitate reproducible research, (3) a graphical data analysis and programming tool that visualizes all aspects of an IMRT plan including dose, contour, and image data to aid the analysis of treatment plans, and (4) a software package that calculates radiobiological models to evaluate IMRT treatment plans. Given the limited number of general-purpose computational environments for radiotherapy research and outcome studies, this computational platform represents a powerful and convenient tool that is well suited for analyzing dose distributions biologically and correlating them with the delivered radiation dose distributions and other patient-related clinical factors. In addition the database is web-based and accessible by multiple users, facilitating its convenient application and use. PMID- 19544787 TI - Investigation of lung nodule detectability in low-dose 320-slice computed tomography. AB - Low-dose imaging protocols in chest CT are important in the screening and surveillance of suspicious and indeterminate lung nodules. Techniques that maintain nodule detectability yet permit dose reduction, particularly for large body habitus, were investigated. The objective of this study was to determine the extent to which radiation dose can be minimized while maintaining diagnostic performance through knowledgeable selection of reconstruction techniques. A 320 slice volumetric CT scanner (Aquilion ONE, Toshiba Medical Systems) was used to scan an anthropomorphic phantom at doses ranging from approximately 0.1 mGy up to that typical of low-dose CT (LDCT, approximately 5 mGy) and diagnostic CT (approximately 10 mGy). Radiation dose was measured via Farmer chamber and MOSFET dosimetry. The phantom presented simulated nodules of varying size and contrast within a heterogeneous background, and chest thickness was varied through addition of tissue-equivalent bolus about the chest. Detectability of a small solid lung nodule (3.2 mm diameter, -37 HU, typically the smallest nodule of clinical significance in screening and surveillance) was evaluated as a function of dose, patient size, reconstruction filter, and slice thickness by means of nine-alternative forced-choice (9AFC) observer tests to quantify nodule detectability. For a given reconstruction filter, nodule detectability decreased sharply below a threshold dose level due to increased image noise, especially for large body size. However, nodule detectability could be maintained at lower doses through knowledgeable selection of (smoother) reconstruction filters. For large body habitus, optimal filter selection reduced the dose required for nodule detection by up to a factor of approximately 3 (from approximately 3.3 mGy for sharp filters to approximately 1.0 mGy for the optimal filter). The results indicate that radiation dose can be reduced below the current low-dose (5 mGy) and ultralow-dose (1 mGy) levels with knowledgeable selection of reconstruction parameters. Image noise, not spatial resolution, was found to be the limiting factor in detection of small lung nodules. Therefore, the use of smoother reconstruction filters may permit lower-dose protocols without trade-off in diagnostic performance. PMID- 19544788 TI - Characterization of a water-equivalent fiber-optic coupled dosimeter for use in diagnostic radiology. AB - This work reports on the characterization of a new fiber-optic coupled (FOC) dosimeter for use in the diagnostic radiology energy range. The FOC dosimeter was constructed by coupling a small cylindrical plastic scintillator, 500 microm in diameter and 2 mm in length, to a 2 m long optical fiber, which acts as a light guide to transmit scintillation photons from the sensitive element to a photo multiplier tube (PMT). A serial port interface on the PMT permits real-time monitoring of light output from the dosimeter via a custom computer program. The FOC dosimeter offered excellent sensitivity and reproducibility, allowing doses as low as 0.16 mGy to be measured with a coefficient of variation of only 3.64%. Dose linearity was also excellent with a correlation coefficient of 1.000 over exposures ranging from 0.16 to 57.29 mGy. The FOC dosimeter exhibited little angular dependence from axial irradiation, varying by less than 5% over an entire revolution. A positive energy dependence was observed and measurements performed within a scatter medium yielded a 10% variation in sensitivity as beam quality changed due to hardening and scatter across a 16 cm depth range. The current dosimetry system features an array of five PMTs to allow multiple FOC dosimeters to be monitored simultaneously. Overall, the system allows for rapid and accurate dose measurements relevant to a range of diagnostic imaging applications. PMID- 19544789 TI - Closed bore XMR (CBXMR) systems for aortic valve replacement: active magnetic shielding of x-ray tubes. AB - Hybrid closed bore x-ray/MRI systems are being developed to improve the safety and efficacy of percutaneous aortic valve replacement procedures by harnessing the complementary strengths of the x-ray and MRI modalities in a single interventional suite without requiring patient transfer between two rooms. These systems are composed of an x-ray C-arm in close proximity (approximately 1 m) to an MRI scanner. The MRI magnetic fringe field can cause the electron beam in the x-ray tube to deflect. The deflection causes the x-ray field of view to shift position on the detector receptacle. This could result in unnecessary radiation exposure to the patient and the staff in the cardiac catheterization laboratory. Therefore, the electron beam deflection must be corrected. The authors developed an active magnetic shielding system that can correct for electron beam deflection to within an accuracy of 5% without truncating the field of view or increasing exposure to the patient. This system was able to automatically adjust to different field strengths as the external magnetic field acting on the x-ray tube was changed. Although a small torque was observed on the shielding coils of the active shielding system when they were placed in a magnetic field, this torque will not impact their performance if they are securely mounted on the x-ray tube and the C-arm. The heating of the coils of the shielding system for use in the clinic caused by electric current was found to be slow enough not to require a dedicated cooling system for one percutaneous aortic valve replacement procedure. However, a cooling system will be required if multiple procedures are performed in one session. PMID- 19544790 TI - On-board SPECT for localizing functional targets: a simulation study. AB - Single photon emission computed tomography (SPECT) was investigated for imaging on-board radiation therapy machines in order to localize functional and molecular targets. A computer-simulated female NCAT phantom was positioned supine on a flat top treatment couch. Twenty tumor locations were defined in the upper torso. The eight lung tumors were subject to the effects of respiratory motion. Tumor diameters of 10.8, 14.4, and 21.6 mm were simulated for tumor-to-background ratios of 3:1 and 6:1 that are characteristic of the radiotracer 99mTc-sestamibi. Projection images representing scan times of 4, 8, and 20 min were simulated for an anterior, half-circular trajectory. Images were reconstructed with attenuation correction by ordered-subsets expectation maximization (OSEM) using six subsets and five iterations. Contrast-to-noise ratios (CNRs) were calculated from ensembles of 25 images. Cross correlation with a noise-free tumor template was used to select the most suspicious tumor location within a 14.4-mm-radius search volume surrounding each tumor, with only that one tumor in each search volume. Localization accuracy was assessed by calculating average distances between measured and true tumor locations. Localization accuracy and CNRs were strongly affected by tumor location relative to the detector trajectory. For example, CNR values near the chest wall were greater by a factor of 3.5 than for tumors near the spine and posterior ribs, a much greater effect than the factor of 1.6 difference in CNR between 6:1 and 3:1 tumor uptakes. Typically, tumors of 6:1 uptake were localized as accurately with 4 min of scan time as tumors of 3:1 uptake that had been imaged for 20 min. Using 4 min scans, 14.4 and 21.6 mm anterior tumors of 6:1 uptake were localized within 2 mm. These results suggest that SPECT, on-board radiation therapy machines, may be a viable modality for localizing certain functional and molecular targets using relatively short scan times. PMID- 19544791 TI - Exploration of the potential of liquid scintillators for real-time 3D dosimetry of intensity modulated proton beams. AB - In this study, the authors investigated the feasibility of using a 3D liquid scintillator (LS) detector system for the verification and characterization of proton beams in real time for intensity and energy-modulated proton therapy. A plastic tank filled with liquid scintillator was irradiated with pristine proton Bragg peaks. Scintillation light produced during the irradiation was measured with a CCD camera. Acquisition rates of 20 and 10 frames per second (fps) were used to image consecutive frame sequences. These measurements were then compared to ion chamber measurements and Monte Carlo simulations. The light distribution measured from the images acquired at rates of 20 and 10 fps have standard deviations of 1.1% and 0.7%, respectively, in the plateau region of the Bragg curve. Differences were seen between the raw LS signal and the ion chamber due to the quenching effects of the LS and due to the optical properties of the imaging system. The authors showed that this effect can be accounted for and corrected by Monte Carlo simulations. The liquid scintillator detector system has a good potential for performing fast proton beam verification and characterization. PMID- 19544792 TI - A method to dynamically balance intensity modulated radiotherapy dose between organs-at-risk. AB - The IMRT treatment planning process typically follows a path that is based on the manner in which the planner interactively adjusts the target and organ-at-risk (OAR) constraints and priorities. The time-intensive nature of this process restricts the planner from fully understanding the dose tradeoff between structures, making it unlikely that the resulting plan fully exploits the extent to which dose can be redistributed between anatomical structures. Multiobjective Pareto optimization has been used in the past to enable the planner to more thoroughly explore alternatives in dose trade-off by combining pre-generated Pareto optimal solutions in real time, thereby potentially tailoring a plan more exactly to requirements. However, generating the Pareto optimal solutions can be nonintuitive and computationally time intensive. The author presents an intuitive and fast non-Pareto approach for generating optimization sequences (prior to planning), which can then be rapidly combined by the planner in real time to yield a satisfactory plan. Each optimization sequence incrementally reduces dose to one OAR at a time, starting from the optimization solution where dose to all OARs are reduced with equal priority, until user-specified target coverage limits are violated. The sequences are computationally efficient to generate, since the optimization at each position along a sequence is initiated from the end result of the previous position in the sequence. The pre-generated optimization sequences require no user interaction. In real time, a planner can more or less instantaneously visualize a treatment plan by combining the dose distributions corresponding to user-selected positions along each of the optimization sequences (target coverage is intrinsically maintained in the combination). Interactively varying the selected positions along each of the sequences enables the planner to rapidly understand the nature of dose trade-off between structures and, thereby, arrive at a suitable plan in a short time. This methodology is demonstrated on a prostate cancer case and olfactory neuroblastoma case. PMID- 19544793 TI - Comparison of power spectra for tomosynthesis projections and reconstructed images. AB - Burgess et al. [Med. Phys. 28, 419-437 (2001)] showed that the power spectrum of mammographic breast background follows a power law and that lesion detectability is affected by the power-law exponent beta which measures the amount of structure in the background. Following the study of Burgess et al., the authors measured and compared the power-law exponent of mammographic backgrounds in tomosynthesis projections and reconstructed slices to investigate the effect of tomosynthesis imaging on background structure. Our data set consisted of 55 patient cases. For each case, regions of interest (ROIs) were extracted from both projection images and reconstructed slices. The periodogram of each ROI was computed by taking the squared modulus of the Fourier transform of the ROI. The power-law exponent was determined for each periodogram and averaged across all ROIs extracted from all projections or reconstructed slices for each patient data set. For the projections, the mean beta averaged across the 55 cases was 3.06 (standard deviation of 0.21), while it was 2.87 (0.24) for the corresponding reconstructions. The difference in beta for a given patient between the projection ROIs and the reconstructed ROIs averaged across the 55 cases was 0.194, which was statistically significant (p < 0.001). The 95% CI for the difference between the mean value of beta for the projections and reconstructions was [0.170, 0.218]. The results are consistent with the observation that the amount of breast structure in the tomosynthesis slice is reduced compared to projection mammography and that this may lead to improved lesion detectability. PMID- 19544794 TI - Dosimetric verification of a commercial Monte Carlo treatment planning system (VMC++) for a 9 MeV electron beam. AB - The aim of this work was to evaluate the performance of the voxel-based Monte Carlo algorithm implemented in the commercial treatment planning system ONCENTRA MASTERPLAN for a 9 MeV electron beam produced by a linear accelerator Varian Clinac 2100 C/D. In order to realize an experimental verification of the computed data, three different groups of tests were planned. The first set was performed in a water phantom to investigate standard fields, custom inserts, and extended treatment distances. The second one concerned standard field, irregular entrance surface, and oblique incidence in a homogeneous PMMA phantom. The last group involved the introduction of inhomogeneities in a PMMA phantom to simulate high and low density materials such as bone and lung. Measurements in water were performed by means of cylindrical and plane-parallel ionization chambers, whereas measurements in PMMA were carried out by the use of radiochromic films. Point dose values were compared in terms of percentage difference, whereas the gamma index tool was used to perform the comparison between computed and measured dose profiles, considering different tolerances according to the test complexity. In the case of transverse scans, the agreement was searched in the plane formed by the intersection of beam axis and the profile (2D analysis), while for percentage depth dose curves, only the beam axis was explored (1D analysis). An excellent agreement was found for point dose evaluation in water (discrepancies smaller than 2%). Also the comparison between planned and measured dose profiles in homogeneous water and PMMA phantoms showed good results (agreement within 2%-2 mm). Profile evaluation in phantoms with internal inhomogeneities showed a good agreement in the case of "lung" insert, while in tests concerning a small "bone" inhomogeneity, a discrepancy was particularly evidenced in dose values on the beam axis. This is due to the inaccurate geometrical description of the phantom that is linked to the calculation voxel size, a feature over which the user has no control. PMID- 19544795 TI - Measurement of relative output factors for the 8 and 4 mm collimators of Leksell Gamma Knife Perfexion by film dosimetry. AB - Three types of films, Kodak EDR2, Gafchromic EBT, and Gafchromic MD-V2-55, were used to measure relative output factors of 4 and 8 mm collimators of the Leksell Gamma Knife Perfexion. The optical density to dose calibration curve for each of the film types was obtained by exposing the films to a range of known doses. Ten data points were acquired for each of the calibration curves in the dose ranges from 0 to 4 Gy, 0 to 8 Gy, and 0 to 80 Gy for Kodak EDR2, Gafchromic EBT, and Gafchromic MD-V2-55 films, respectively. For the measurement of relative output factors, five films of each film type were exposed to a known dose. All films were scanned using EPSON EXPRESSION 10000 XL scanner with 200 dpi resolution in 16 bit gray scale for EDR2 film and 48 bit color scale for Gafchromic films. The scanned images were imported in the red channel for both Gafchromic films. The background corrections from an unexposed film were applied to all films. The output factors obtained from film measurements were in a close agreement both with the Monte Carlo calculated values of 0.924 and 0.805 for 8 and 4 mm collimators, respectively. These values are provided by the vendor and used as default values in the vendor's treatment planning system. The largest differences were noted for the Kodak EDR 2 films (-2.1% and -4.5% for 8 and 4 mm collimators, respectively). The best agreement observed was for EBT Gafchromic film (-0.8% and +0.6% differences for 8 and 4 mm collimators, respectively). Based on the present values, no changes in the default relative output factor values were made in the treatment planning system. PMID- 19544796 TI - Optimal number of respiratory gates in positron emission tomography: a cardiac patient study. AB - Respiratory gating is the method of dividing the data from a tomographic scan with respect to the respiratory phase of the patient. It enables more accurate images by reducing the effects of motion blur and attenuation artifacts due to motion. However, it induces image degradation due to higher noise levels as the number of events per gate is reduced. Due to lack of systematic studies in this regard, different numbers of gates are being used in the scientific and clinical practice. The present study aims at examining the relationship between the respiratory signal, the number of gates required for accurate motion detection, and the level of noise with two different methods of gating: (1) Amplitude-based gating and (2) time-based gating. Patient data with a wide range of motion are used for the study. The results show that time-based gating underestimates the real respiratory displacement by up to 50%. The optimal number of gates is 8 for amplitude- and 6 for time-based gatings. The noise properties remain the same with either method but noise increases with increasing number of gates. PMID- 19544797 TI - Systematic uncertainties in the Monte Carlo calculation of ion chamber replacement correction factors. AB - In a previous study [Med. Phys. 35, 1747-1755 (2008)], the authors proposed two direct methods of calculating the replacement correction factors (P(repl) or P(cav)P(dis)) for ion chambers by Monte Carlo calculation. By "direct" we meant the stopping-power ratio evaluation is not necessary. The two methods were named as the high-density air (HDA) and low-density water (LDW) methods. Although the accuracy of these methods was briefly discussed, it turns out that the assumption made regarding the dose in an HDA slab as a function of slab thickness is not correct. This issue is reinvestigated in the current study, and the accuracy of the LDW method applied to ion chambers in a 60Co photon beam is also studied. It is found that the two direct methods are in fact not completely independent of the stopping-power ratio of the two materials involved. There is an implicit dependence of the calculated P(repl) values upon the stopping-power ratio evaluation through the choice of an appropriate energy cutoff delta, which characterizes a cavity size in the Spencer-Attix cavity theory. Since the delta value is not accurately defined in the theory, this dependence on the stopping power ratio results in a systematic uncertainty on the calculated P(repl) values. For phantom materials of similar effective atomic number to air, such as water and graphite, this systematic uncertainty is at most 0.2% for most commonly used chambers for either electron or photon beams. This uncertainty level is good enough for current ion chamber dosimetry, and the merits of the two direct methods of calculating P(repl) values are maintained, i.e., there is no need to do a separate stopping-power ratio calculation. For high-Z materials, the inherent uncertainty would make it practically impossible to calculate reliable P(repl) values using the two direct methods. PMID- 19544798 TI - Improved critical structure sparing with biologically based IMRT optimization. AB - The impact of using biological models in treatment planning on plan quality is studied by comparing IMRT plans generated using selected commercially available treatment planning systems (TPSs) employing biological models/quantities in IMRT optimization (bIMRT) and the conventional physically (dose-volume) based optimization (pIMRT). A total of 25 IMRT plans, generated for five cases of different anatomic sites (brain, head and neck, lung, pancreas, and prostate) using five TPSs, two bIMRT (CMS Monaco and Phillips Pinnacle3 P3IMRT) and three pIMRT (CMS Xio, Phillips Pinnacle3, and Tomotherapy) systems, were compared. Dose volume histograms, maximum, minimum, and mean doses, target heterogeneity and conformity indices, equivalent uniform dose (EUD), and an overall plan-ranking index (fEUD) were used in the comparison. It is clear from the comparison that the use of biological models in treatment planning optimization can generate IMRT plans with significantly improved normal tissue sparing with similar or slightly increased dose heterogeneity in the target, as compared to the conventional dose volume based optimization for the same beam arrangement. For example, the bIMRT plans lead to smaller EUDs in 32 out of 37 normal structures in all five cases combined, as compared to the pIMRT plans. Caution should be exercised in choosing appropriate models and/or model parameters and in evaluating the plan obtained when using the biologically based treatment planning system. PMID- 19544800 TI - Novel dosimetric phantom for quality assurance of volumetric modulated arc therapy. AB - The objective of this work is to assess the suitability and performance of a new dosimeter system with a novel geometry for the quality assurance (QA) of volumetric modulated arc therapy (VMAT). The new dosimeter system consists of a hollow cylinder (15 and 25 cm inner and outer diameters) with 124 diodes embedded in the phantom's cylindrical wall forming four rings of detectors. For coplanar beams, the cylindrical geometry and the ring diode pattern offer the advantage of invariant perpendicular incidence on the beam central axis for any gantry angle and also have the benefit of increasing the detector density as both walls of the cylinder sample the beam. Other advantages include real-time readout and reduced weight with the hollow phantom shape. A calibration method taking into account the variation in radiation sensitivity of the diodes as a function of gantry angle was developed and implemented. In this work, the new dosimeter system was used in integrating mode to perform composite dose measurements along the cylindrical surface supporting the diodes. The reproducibility of the dosimeter response and the angular dependence of the diodes were assessed using simple 6 MV photon static beams. The performance of the new dosimeter system for VMAT QA was then evaluated using VMAT plans designed for a head and neck, an abdominal sarcoma, and a prostate patient. These plans were optimized with 90 control points (CPs) and additional versions of each plan were generated by increasing the number of CPs to 180 and 360 using linear interpolation. The relative dose measured with the dosimeter system for the VMAT plans was compared to the corresponding TPS dose map in terms of relative dose difference (% deltaD) and distance to agreement (DTA). The dosimeter system's sensitivity to gantry rotation offset and scaling errors as well as setup errors was also evaluated. For static beams, the dosimeter system offered good reproducibility and demonstrated small residual diode angular dependence after calibration. For VMAT deliveries, the agreement between measured and calculated doses was good with > or = 86.4% of the diodes satisfying 3% of % deltaD or 2 mm DTA for the 180 CP plans. The phantom offered sufficient sensitivity for the detection of small gantry rotation offset (3 degrees) and scaling errors (1 degree) as well as phantom setup errors of 1 mm, although the results were plan dependent. With its novel geometry, the dosimeter system was also able to experimentally demonstrate the discretization effect of the number of CPs used in the TPS to simulate a continuous arc. These results demonstrate the suitability of the new dosimeter system for the patient-specific QA of VMAT plans and suggest that the dosimeter system can be an effective tool in the routine QA and commissioning of treatment machines capable of VMAT delivery and cone-beam CT image guidance. PMID- 19544799 TI - Automatic image-to-world registration based on x-ray projections in cone-beam CT guided interventions. AB - Intraoperative imaging offers a means to account for morphological changes occurring during the procedure and resolve geometric uncertainties via integration with a surgical navigation system. Such integration requires registration of the image and world reference frames, conventionally a time consuming, error-prone manual process. This work presents a method of automatic image-to-world registration of intraoperative cone-beam computed tomography (CBCT) and an optical tracking system. Multimodality (MM) markers consisting of an infrared (IR) reflective sphere with a 2 mm tungsten sphere (BB) placed precisely at the center were designed to permit automatic detection in both the image and tracking (world) reference frames. Image localization is performed by intensity thresholding and pattern matching directly in 2D projections acquired in each CBCT scan, with 3D image coordinates computed using backprojection and accounting for C-arm geometric calibration. The IR tracking system localized MM markers in the world reference frame, and the image-to-world registration was computed by rigid point matching of image and tracker point sets. The accuracy and reproducibility of the automatic registration technique were compared to conventional (manual) registration using a variety of marker configurations suitable to neurosurgery (markers fixed to cranium) and head and neck surgery (markers suspended on a subcranial frame). The automatic technique exhibited subvoxel marker localization accuracy (< 0.8 mm) for all marker configurations. The fiducial registration error of the automatic technique was (0.35 +/-0.01) mm, compared to (0.64 +/- 0.07 mm) for the manual technique, indicating improved accuracy and reproducibility. The target registration error (TRE) averaged over all configurations was 1.14 mm for the automatic technique, compared to 1.29 mm for the manual in accuracy, although the difference was not statistically significant (p = 0.3). A statistically significant improvement in precision was observed-specifically, the standard deviation in TRE was 0.2 mm for the automatic technique versus 0.34 mm for the manual technique (p = 0.001). The projection based automatic registration technique demonstrates accuracy and reproducibility equivalent or superior to the conventional manual technique for both neurosurgical and head and neck marker configurations. Use of this method with C arm CBCT eliminates the burden of manual registration on surgical workflow by providing automatic registration of surgical tracking in 3D images within approximately 20 s of acquisition, with registration automatically updated with each CBCT scan. The automatic registration method is undergoing integration in ongoing clinical trials of intraoperative CBCT-guided head and neck surgery. PMID- 19544801 TI - Magnetic nanoparticle temperature estimation. AB - The authors present a method of measuring the temperature of magnetic nanoparticles that can be adapted to provide in vivo temperature maps. Many of the minimally invasive therapies that promise to reduce health care costs and improve patient outcomes heat tissue to very specific temperatures to be effective. Measurements are required because physiological cooling, primarily blood flow, makes the temperature difficult to predict a priori. The ratio of the fifth and third harmonics of the magnetization generated by magnetic nanoparticles in a sinusoidal field is used to generate a calibration curve and to subsequently estimate the temperature. The calibration curve is obtained by varying the amplitude of the sinusoidal field. The temperature can then be estimated from any subsequent measurement of the ratio. The accuracy was 0.3 degree K between 20 and 50 degrees C using the current apparatus and half-second measurements. The method is independent of nanoparticle concentration and nanoparticle size distribution. PMID- 19544802 TI - Virtual tagging for laxative-free CT colonography: pilot evaluation. AB - Laxative-free computed tomographic colonography (lfCTC) could significantly improve patient adherence to colorectal screening. However, the interpretation of lfCTC data is complicated by the presence of poorly tagged feces and partial volume artifacts that imitate colorectal lesions. The authors developed a method for virtual tagging of such artifacts. A probabilistic model of colonic wall was developed, and virtual tagging was performed on artifacts that were identified by the model. The method was evaluated with 46 clinical lfCTC cases that were prepared with dietary fecal tagging only. Visual examples show that the method can label partial-volume artifacts, poorly tagged feces, nonadhering completely untagged feces, and artifacts such as rectal tubes. The effect of virtual tagging was evaluated by comparing the detection accuracy of a fully automated polyp detection scheme without and with the method. With virtual tagging, the per lesion detection sensitivity was 100% for lesions > or = 10 mm (n = 4) with 3.8 false positives per patient (per two CT scan volumes) and 90% for lesions > or = 6 mm (n = 10) with 5.4 false positives per patient on average. The improvement in detection performance by virtual tagging was statistically significant (p = 0.03; JAFROC and JAFROC-1). PMID- 19544803 TI - Pseudomonoenergetic x-ray diffraction measurements using balanced filters for coherent-scatter computed tomography. AB - Coherent-scatter computed tomography (CSCT) is a method of "composition" imaging based on measurements of diffraction patterns from tissues. Use of an x-ray tube degrades scatter pattern angular resolution due to the x-ray spectral width, making it difficult to uniquely identify some materials. The use of two transmission filters with similar atomic numbers (balanced "Ross filters") to generate pseudomonoenergetic scatter patterns is described as it applies to CSCT. An analysis of angular-blur mechanisms reveals that focal spot size and beam width are the most important factors determining Bragg-peak width when Er-Tm filters are used. A relative RMS spectral width of 1% can be achieved in the difference spectrum and a Bragg-peak RMS angular width of approximately 0.14 degree (relative width of 3% at 5 degrees scatter angle) can be achieved with an effective energy of 58 keV. PMID- 19544804 TI - Monte Carlo study of LDR seed dosimetry with an application in a clinical brachytherapy breast implant. AB - A Monte Carlo (MC) study was carried out to evaluate the effects of the interseed attenuation and the tissue composition for two models of 125I low dose rate (LDR) brachytherapy seeds (Medi-Physics 6711, IBt InterSource) in a permanent breast implant. The effect of the tissue composition was investigated because the breast localization presents heterogeneities such as glandular and adipose tissue surrounded by air, lungs, and ribs. The absolute MC dose calculations were benchmarked by comparison to the absolute dose obtained from experimental results. Before modeling a clinical case of an implant in heterogeneous breast, the effects of the tissue composition and the interseed attenuation were studied in homogeneous phantoms. To investigate the tissue composition effect, the dose along the transverse axis of the two seed models were calculated and compared in different materials. For each seed model, three seeds sharing the same transverse axis were simulated to evaluate the interseed effect in water as a function of the distance from the seed. A clinical study of a permanent breast 125I implant for a single patient was carried out using four dose calculation techniques: (1) A TG-43 based calculation, (2) a full MC simulation with realistic tissues and seed models, (3) a MC simulation in water and modeled seeds, and (4) a MC simulation without modeling the seed geometry but with realistic tissues. In the latter, a phase space file corresponding to the particles emitted from the external surface of the seed is used at each seed location. The results were compared by calculating the relevant clinical metrics V85, V100, and V200 for this kind of treatment in the target. D90 and D50 were also determined to evaluate the differences in dose and compare the results to the studies published for permanent prostate seed implants in literature. The experimental results are in agreement with the MC absolute doses (within 5% for EBT Gafchromic film and within 7% for TLD-100). Important differences between the dose along the transverse axis of the seed in water and in adipose tissue are obtained (10% at 3.5 cm). The comparisons between the full MC and the TG-43 calculations show that there are no significant differences for V85 and V100. For V200, 8.4% difference is found coming mainly from the tissue composition effect. Larger differences (about 10.5% for the model 6711 seed and about 13% for the InterSource125) are determined for D90 and D50. These differences depend on the composition of the breast tissue modeled in the simulation. A variation in percentage by mass of the mammary gland and adipose tissue can cause important differences in the clinical dose metrics V200, D90, and D50. Even if the authors can conclude that clinically, the differences in V85, V100, and V200 are acceptable in comparison to the large variation in dose in the treated volume, this work demonstrates that the development of a MC treatment planning system for LDR brachytherapy will improve the dose determination in the treated region and consequently the dose outcome relationship, especially for the skin toxicity. PMID- 19544805 TI - Electron beam quality control using an amorphous silicon EPID. AB - An amorphous silicon EPID has been investigated to determine whether it is capable of quality control constancy measurements for linear accelerator electron beams. The EPID grayscale response was found to be extremely linear with dose over a wide dose range and, more specifically, for exposures of 95-100 MU. Small discrepancies of up to 0.8% in linearity were found at 6 MeV (8-15 MeV showed better agreement). The shape of the beam profile was found to be significantly altered by scatter in air over the approximately 60 cm gap between the end of the applicator and the EPID. Nevertheless, relative changes in EPID-measured profile flatness and symmetry were linearly related to changes in these parameters at 95 cm focus to surface distance (FSD) measured using a 2D diode array. Similar results were obtained at 90 degrees and 270 degrees gantry angles. Six months of daily images were acquired and analyzed to determine whether the device is suitable as a constancy checker. EPID output measurements agreed well with daily ion chamber measurements, with a 0.8% standard deviation in the difference between the two measurement sets. When compared to weekly parallel plate chamber measurements, this figure dropped to 0.5%. A Monte Carlo (MC) model of the EPID was created and demonstrated excellent agreement between MC-calculated profiles in water and the EPID at 95 and 157 cm FSD. Good agreement was also found with measured EPID profiles, demonstrating that the EPID provides an accurate measurement of electron profiles. The EPID was thus shown to be an effective method for performing electron beam daily constancy checks. PMID- 19544807 TI - The use of novel gradient directions with DTI to synthesize data with complicated diffusion behavior. AB - This study demonstrates a new technique for synthesizing diffusion tensor imaging (DTI) data sets that exhibit complex diffusion characteristics by performing operations on acquired DTI data of simple structures with anisotropic diffusive properties. The motivation behind this technique is to characterize the behavior of noise in complicated data using a phantom. Compared to simulations, an advantage to this approach is that the acquired data contain noise characteristic of the scanner and protocol. Using this technique, a simple capillary phantom is employed to infer the quality of data for more clinically realistic tissue structures (e.g., crossing fiber tracts). A water-filled phantom containing capillary arrays was constructed to demonstrate this technique, which uses a DTI protocol with typical clinical parameters. Eigenvalues and fractional anisotropy were calculated for the initial prolate data. Data were adjusted to synthesize different apparent diffusion coefficient (ADC) spatial distributions, which were compared to theoretical and analytical models. RMS differences and volumetric overlap between expected and measured ADC distributions were quantified for all synthesized distributions. Differences between synthesized and actual distributions were discussed. PMID- 19544806 TI - An MRI-compatible system for focused ultrasound experiments in small animal models. AB - The development of novel MRI-guided therapeutic ultrasound methods including potentiated drug delivery and targeted thermal ablation requires extensive testing in small animals such as rats and mice due to the widespread use of these species as models of disease. An MRI-compatible, computer-controlled three-axis positioning system was constructed to deliver focused ultrasound exposures precisely to a target anatomy in small animals for high-throughput preclinical drug delivery studies. Each axis was constructed from custom-made nonmagnetic linear ball stages driven by piezoelectric actuators and optical encoders. A range of motion of 5 x 5 x 2.5 cm3 was achieved, and initial bench top characterization demonstrated the ability to deliver ultrasound to the brain with a spatial accuracy of 0.3 mm. Operation of the positioning system within the bore of a clinical 3 T MR imager was feasible, and simultaneous motion and MR imaging did not result in any mutual interference. The system was evaluated in its ability to deliver precise sonications within the mouse brain, linear scanned exposures in a rat brain for blood barrier disruption, and circular scans for controlled heating under MR temperature feedback. Initial results suggest that this is a robust and precise apparatus for use in the investigation of novel ultrasound-based therapeutic strategies in small animal preclinical models. PMID- 19544808 TI - Characterization of a mini-multileaf collimator in a proton beamline. AB - A mini-multileaf collimator (MMLC) was mounted as a field shaping collimator in a proton beamline at the Massachusetts General Hospital. The purpose is to evaluate the device's dosimetric and mechanical properties for the use in a proton beamline. For this evaluation, the authors compared MMLC and brass aperture shaped dose distributions with regard to lateral and depth dose properties. The lateral fall off is generally broader with the MMLC, with difference varying with proton range from 0.2 to 1.2 mm. Central axis depth dose curves did not show a difference in peak-to-entrance ratio, peak width, distal fall off, or range. Two dimensional dose distributions to investigate the conformity of MMLC shaped doses show that the physical leaf width of approximately 2.5 mm does not have a significant impact. All differences seen in dose distribution shaped by the MMLC versus brass apertures were shown to be clinically insignificant. Measured neutron doses of 0.03-0.13 mSv/Gy for a closed brass beam block (depending on range) are very low compared to the previously published data. Irradiation of the tungsten MMLC, however, produced 1.5-1.8 times more neutrons than brass apertures. Exposure of the staff resulting from activation of the device is below regulatory limits. The measurements established an equivalency between aperture and MMLC shaped dose distributions. PMID- 19544809 TI - Search for IMRT inverse plans with piecewise constant fluence maps using compressed sensing techniques. AB - An intensity-modulated radiation therapy (IMRT) field is composed of a series of segmented beams. It is practically important to reduce the number of segments while maintaining the conformality of the final dose distribution. In this article, the authors quantify the complexity of an IMRT fluence map by introducing the concept of sparsity of fluence maps and formulate the inverse planning problem into a framework of compressing sensing. In this approach, the treatment planning is modeled as a multiobjective optimization problem, with one objective on the dose performance and the other on the sparsity of the resultant fluence maps. A Pareto frontier is calculated, and the achieved dose distributions associated with the Pareto efficient points are evaluated using clinical acceptance criteria. The clinically acceptable dose distribution with the smallest number of segments is chosen as the final solution. The method is demonstrated in the application of fixed-gantry IMRT on a prostate patient. The result shows that the total number of segments is greatly reduced while a satisfactory dose distribution is still achieved. With the focus on the sparsity of the optimal solution, the proposed method is distinct from the existing beamlet- or segment-based optimization algorithms. PMID- 19544810 TI - Focused RF hyperthermia using magnetic fluids. AB - Heat therapies such as hyperthermia and thermoablation are very promising approaches in the treatment of cancer. Compared with available hyperthermia modalities, magnetic fluid hyperthermia (MFH) yields better results in uniform heating of the deeply situated tumors. In this approach, fluid consisting of superparamagnetic particles (magnetic fluid) is delivered to the tumor. An alternating (ac) magnetic field is then used to heat the particles and the corresponding tumor, thereby ablating it. However, one of the most serious shortcomings of this technique is the unwanted heating of the healthy tissues. This results from the magnetic fluid diffusion from the tumor to the surrounding tissues or from incorrect localization of the fluids in the target tumor area. In this study, the authors demonstrated that by depositing appropriate static (dc) magnetic field gradients on the alternating (ac) magnetic fields, focused heating of the magnetic particles can be achieved. A focused hyperthermia system was implemented by using two types of coils: dc and ac coils. The ac coil generated the alternating magnetic field responsible for the heating of the magnetic particles; the dc coil was used to superimpose a static magnetic field gradient on the alternating magnetic field. In this way, focused heating of the particles was obtained in the regions where the static field was dominated by the alternating magnetic field. In vitro experiments showed that as the magnitude of the dc solenoid currents was increased from 0 to 1.8 A, the specific absorption rate (SAR) of the superparamagnetic particles 2 cm apart from the ac solenoid center decreased by a factor of 4.5, while the SAR of the particles at the center was unchanged. This demonstrates that the hyperthermia system is capable of precisely focusing the heat at the center. Additionally, with this approach, shifting of the heat focus can be achieved by applying different amounts of currents to individual dc solenoids. In vivo experiments were performed with adult rats, where magnetic fluids were injected percutaneously into the tails (with homogeneous fluid distribution inside the tails). Histological examination showed that, as we increased the dc solenoid current from 0.5 to 1.8 A, the total burned volume decreased from 1.6 to 0.2 cm3 verifying the focusing capability of the system. The authors believe that the studies conducted in this work show that MFH can be a much more effective method with better heat localization and focusing abilities. PMID- 19544811 TI - A flexible multicamera visual-tracking system for detecting and correcting motion induced artifacts in cardiac SPECT slices. AB - Patient motion is inevitable in SPECT and PET due to the lengthy period of time patients are imaged. The authors hypothesized that the use of external-tracking devices which provide additional information on patient motion independent of SPECT data could be employed to provide a more robust correction than obtainable from data-driven methods. Therefore, the authors investigated the Vicon MX visual tracking system (VTS) which utilizes near-infrared (NIR) cameras to stereo-image small retroreflective markers on stretchy bands wrapped about the chest and abdomen of patients during cardiac SPECT. The chest markers are used to provide an estimate of the rigid-body (RB) motion of the heart. The abdomen markers are used to provide a signal used to bin list-mode acquisitions as part of correction of respiratory motion of the heart. The system is flexible in that the layout of the cameras can be designed to facilitate marker viewing. The system also automatically adapts marker tracking to employ all of the cameras visualizing a marker at any instant, with visualization by any two being sufficient for stereo tracking. Herein the ability of this VTS to track motion with submillimeter and subdegree accuracy is established through studies comparing the motion of Tc-99m containing markers as assessed via stereo-tracking and from SPECT reconstructions. The temporal synchronization between motion-tracking data and timing marks embedded in list-mode SPECT acquisitions is shown to agree within 100 ms. In addition, motion artifacts were considerably reduced in reconstructed SPECT slices of an anthropomorphic phantom by employing within iterative reconstruction the motion-tracking information from markers attached to the phantom. The authors assessed the number and placement of NIR cameras required for robust motion tracking of markers during clinical imaging in 77 SPECT patients. They determined that they were able to track without loss during the entire period of SPECT and transmission imaging at least three of the four markers on the chest and one on the abdomen bands 94% and 92% of the time, respectively. The ability of the VTS to correct motion clinically is illustrated for ten patients who volunteered to undergo repeat-rest imaging with the original rest SPECT study serving as the standard against which to compare the success of correction. Comparison of short-axis slices shows that VTS-based motion correction provides better agreement with the original-rest-imaging slices than either no correction or the vendor-supplied software for motion correction on, our SPECT system. Comparison of polar maps shows that VTS-based motion-correction results in less numerical difference on average in the segments of the polar maps between the original-rest study and the second-rest study than the other two strategies. The difference was statistically significant for the comparison between VTS-based and clinical vendor-supplied software correction. Taken together, these findings suggest that VTS-based motion correction is superior to either no-motion correction or the vendor-supplied software the authors investigated in clinical practice. PMID- 19544812 TI - Secondary caries, a problem of primary concern. PMID- 19544813 TI - Buonocore Memorial Lecture. Dental erosion. AB - There is some evidence that dental erosion is steadily spreading. To diagnose erosion, dental professionals have to rely on clinical appearance, as there is no device available to detect it. Adequate preventive measures can only be initiated if the different risk factors and potential interactions between them are known. When substance loss, caused by erosive tooth wear, reaches a certain degree, oral rehabilitation becomes necessary. Prior to the most recent decade, the severely eroded dentition could only be rehabilitated by the provision of extensive crown and bridgework or removable dentures. As a result of the improvements in composite restorative materials and in adhesive techniques, it has become possible to rehabilitate eroded dentitions in a less invasive manner. PMID- 19544814 TI - Clinical evaluation of ceramic inlays compared to composite restorations. AB - This study compared the clinical performance of indirectly manufactured ceramic Evopress inlays with those of directly placed, fine particle hybrid Filtek Z250 composite restorations in posterior teeth. From January 2000 to October 2003, 109 patients received 264 Evopress (Wegold) ceramic inlays and 68 patients received 145 Filtek Z250 (3M ESPE) composite restorations in a dental office. Two-hundred and fifty ceramic inlays (95%) and 135 composite restorations (93%) were re examined up to 57 months after placement. Modified USPHS criteria were used for the study. The worst finding of all the assessments was the overall assessment of individual restorations. On the basis of these criteria, 220 (88%) Evopress ceramic inlays were assessed as Alpha at the time of clinical re-examination, 26 (10%) were judged Beta and four ceramic inlays (2%) were rated Delta in the re examination interval and thus categorized as failures. At the time of re examination, 91 of the 135 composite restorations (67%) were judged Alpha, 36 restorations (26%) were rated Beta and three restorations (2%) were judged Charlie. Five restorations (4%) were categorized as failures (Delta). In two cases, there were marginal gap formations; there were also two cases of secondary caries after 28 and 35 months, as well as a fracture after 13 months. According to Kaplan and Meier, the survival rate after 57 months was 94% for ceramic inlays and 93% for composite restorations. The log rank test showed no significant differences in the survival curves. The current study showed that indirectly manufactured Evopress ceramic inlays performed better than direct Filtek Z250 composite restorations in marginal adaptation, color match and anatomic form. However, with regard to survival probability, there was no significant difference. PMID- 19544815 TI - Effect of lateral excursive movements on the progression of abfraction lesions. AB - The theory of abfraction suggests that tooth flexure arising from occlusal loads causes the formation and progression of abfraction lesions. The current study investigated whether reducing occlusal loading by adjusting the occlusion on a tooth during lateral excursive movements had any effect on the rate of progression of existing abfraction lesions. Recruited were 39 subjects who had two non-carious cervical lesions in the maxillary arch that did not need restoration and were in group function during lateral excursive movements of the mandible. One of the teeth was randomly selected to have the excursive occlusal contacts reduced by using a fine grain diamond bur. Centric occlusal contacts were not reduced. Impressions of the lesion were taken over a 30-month period to enable monitoring of the wear rate, and duplicate dies were poured into epoxy resin to allow for sectioning. The size of the lesions was measured using stereomicroscopic analysis of the sectioned epoxy resin dies, and the results were analyzed using an Independent t-test. No statistically significant difference in wear rates between the adjusted and non-adjusted teeth was found (p > 0.05). Within the limitations of the current study, it was concluded that occlusal adjustment does not appear to halt the progression of non-carious cervical lesions; consequently, this procedure cannot be recommended. PMID- 19544816 TI - Bond strength of resin cements to a zirconia ceramic with different surface treatments. AB - This study evaluated the influence of surface treatments and metal primers on the bond strength of resin cements to a yttrium-stabilized tetragonal zirconia (Y TZP) ceramic. Two-hundred and forty plates of Y-TZP ceramic were randomly assigned to 24 groups (n = 10) according to the combination of surface treatment (none, air abrasion with Al2O3 particles, Er:YAG laser irradiation), metal primer (none, Alloy Primer, Metal Primer II or Metaltite) and resin cement (Calibra [Bis GMA-based] or Panavia F2.0 [MDP-based]). Fragments of dentin with a cylindrical edge (0.8 mm in diameter) were fixed to ceramic surfaces with the resin cements. The micro-shear bond test was carried out at a 1 mm/minute speed until failure, and the ceramic surfaces were examined after debonding. Bond strengths were analyzed through three-way ANOVA/Tukey test with a 5% significance level. Changes in topography after surface treatments were evaluated with scanning electron microscopy. Surface treatments significantly modified the topography of the Y-TZP ceramic. Air abrasion resulted in increased bond strength for both resin cements. However, air abraded and laser irradiated specimens presented higher bond strength with the Bis-GMA-based resin cement than with the MDP-based cement. Both cements presented similar behavior on untreated surfaces. The three metal primers yielded a significant increase in bond strength, regardless of the surface treatment and resin cement. Adhesive failures were the most prevalent. Air abrasion with Al2O3 particles and the application of metal primers increased bond strength to Y-TZP surfaces for both resin cements. PMID- 19544817 TI - The effect of three variables on shear bond strength when luting a resin inlay to dentin. AB - The current study evaluated the effects of three variables on the shear bond strength of indirect composite restorations to human dentin. The three variables examined included immediate dentin sealing (IDS), the thinning of dentin adhesives by air-blowing before cementation and light-curing the dentin adhesive before cementation. One-hundred and eighty cylinder composite inlays, 2 mm in diameter and 3 mm in length, were made using a Tescera ATL system (BISCO Inc). Tooth disks 2-mm thick were obtained from 90 freshly-extracted human premolars. Two indirect composite cylinders were assigned to a single tooth disk. The discs were randomly divided into six groups according to the luting methods. AdheSE (Ivoclar Vivadent) was used as the dentin-bonding agent (DBA) for all groups. In Groups 1, 2 and 3, the dentin was sealed with AdheSE before taking the impression. After priming, the adhesive was lightly air-blown, then light-cured. On the other hand, the dentin was not sealed before taking the impression in Groups 4, 5 and 6. Regarding the application of DBA before cementation, it was gently air-blown and light-cured before cementation in Groups 1 and 4; whereas, it was heavily air-blown and light-cured in Groups 2 and 5 and gently air-blown but not light-cured in Groups 3 and 6. Z-250 and Duo-Link were used as luting materials. After 24-hours of storage, the bonded inlays were subjected to a shear bond test. For each luting material, one-way ANOVA and Duncan's Multiple Range Test were used to compare the shear bond strength. Paired t-tests were also performed to compare the shear strength between the two luting materials. All the statistical tests were carried out at the 95% confidence level. In Z-250, the results of the shear bond strength were as follows: Group 1(14.90MPa) > Group 2(12.22MPa), Group 4(12.16MPa) Group 5(9.61MPa), Group 3(9.60MPa) Group 6(3.54MPa)(p<0.05). In Duo-Link, the following shear bond strengths were obtained: Group 1(14.65MPa) > Group 2(13.04MPa), Group 4(12.66MPa) > Group 5(10.10MPa) > Group 3(8.40MPa) > Group 6(2.88MPa) (p<0.05). The mean shear bond strength of Z-250 and Duo-Link were not statistically different with the exception of Group 5. In conclusion, the shear bond strength of the indirect composite restoration to dentin can be improved by dentin sealing with DBA before taking an impression, gently air drying and light curing the DBA before the luting procedure. PMID- 19544818 TI - Influence of the internal conditioning of indirect restorations of resin composite in relation to microleakage using LEDs and QTH units. AB - The current study evaluated in-vitro microleakage of indirect composite inlays fixed with flowable composite and resinous cement and cured by LEDs and QTH units, thus modifying the internal conditioning of the inlays. Thirty-two non carious teeth were selected and 64 cavity preparations were performed in both enamel and cementum. These teeth were divided into four groups: I--Elipar FreeLight appliance and Filtek Flowable composite; II--Ultralux appliance and Filtek Flowable composite; III--Elipar FreeLight appliance and RelyX cement and IV--Ultralux appliance and RelyX cement. The inlays were internally sandblasted with aluminum oxide, etched with 37% phosphoric acid, washed and silanized. The dental etching was carried out with 37% phosphoric acid for 15 seconds in dentin and 30 seconds in enamel. After being washed and dried, Single Bond dentinal adhesive was then applied, and the inlays were fixed with their respective luting agents, cured in close contact with their surfaces for 60 seconds, thermocycled and immersed in 0.5% basic fuchsine solution. The teeth were then washed and sectioned through the center of the restoration in order for the microleakage readings to be performed using the Image Tool Software. Then, ANOVA and Tukey's statistical tests were applied. In enamel, there was no significant difference for both groups with regard to microleakage; in cementum, the significant difference was 5% (p > 0.005). In agreement with the results, the inlays can be fixed with RelyX and Filtek Flowable resins. PMID- 19544819 TI - Effect of filler ratio in adhesive systems on the shear bond strength of resin composite to porcelains. AB - This in vitro study evaluated the effect of six different adhesive systems on the shear bond strength of resin composite to feldspathic and low-fusing porcelains. Sixty porcelain blocks were prepared for each low-fusing (Matchmaker) and feldspathic (MVK95) porcelain specimen. After surface preparation, the porcelain specimens were divided into six groups (n = 10) for different adhesive systems (Adper Prompt L-Pop, QuadrantUnil Bond, Te-Econom, PQ1, One-StepPlus and Prime&Bond NT). After adhesive application, a universal resin composite (FiltekZ250) was condensed on the specimens. The prepared specimens were then stored in distilled water at 37 degrees C for 24 hours, then all the samples were thermal cycled 1000 times between 5 degrees C and 55 degrees C. Shear testing was performed on a universal test machine using a crosshead speed of 0.5 mm/minute. The statistical analysis of the bond strength data included two-way ANOVA. Then, the means were compared by Tukey HSD test (alpha = 0.05). The lowest bond strength was observed in Adper Prompt L-Pop. No statistically significant difference was observed between One-Step Plus and Prime&Bond NT. The highest bond strength was observed in PQ1. When low-fusing or feldspathic porcelain restorations are repaired with resin composite, self-etching adhesive systems may not be indicated. If maximum bond strength is the goal in porcelain resin bonding, adhesive systems that have a high filler ratio should be used. PMID- 19544820 TI - The effect of flowable materials on the microleakage of Class II composite restorations that extend apical to the cemento-enamel junction. AB - This in vitro study investigated the effects of a thin layer of flowable composite or compomer on microleakage occurring in Class II packable and nanofilled composite restorations that extend apical to the cemento-enamel junction (CEJ). The current study also investigated any differences in microleakage that occur between restorations light-cured using a light-emitting diode (LED) and a quartz tungsten halogen (QTH) light-curing unit. Standardized Class II "slot" cavity preparations were prepared on the mesial and distal surfaces of 72 extracted, unrestored, non-carious premolars (n = 144). The gingival margins were placed 1 mm apical to the CEJ. The teeth were divided into two groups (n = 72) and restored with a "packable composite" (Filtek P60) or a "nano-filled composite" (Universal Filtek Supreme XT) with or without flowable composite (Flowable Filtek Supreme XT) or flowable composite (Dyract Flow) as gingival liners placed with thicknesses of 1.0 mm. Each increment was cured for 20 seconds before adding the next. One-half of the samples in each group was cured with QTH (Coltolux 75) and the other half with LED (Coltolux LED) LCUs. After a two-week incubation period at 37 degrees C, the specimens were thermocycled (5 degrees C-55 degrees C x 1500), immersed in 0.5% basic fuchsin dye for 24 hours, sectioned and the microleakage was then evaluated at the gingival margin by two examiners using a 0-3 score scale. Within the current study, when flowable liners were used, both the packable (Filtek P60) and nanofilled (Filtek Supreme XT Universal Restorative) composite materials had significantly less microleakage than when flowable liners were not used (p < 0.05). Both flowable liners (Flowable Filtek Supreme XT and Dyract Flow) resulted in a significant reduction of the microleakage occurring under both types of composite materials at the gingival floors ( p < 0.05), but there was no significant difference between them. The choice of light curing technology (LED vs QTH) had no significant effect on the amount of microleakage observed. PMID- 19544821 TI - Corrosion sealing of amalgam restorations in vitro. AB - Amalgam restorations, when first placed, have been shown to exhibit a gap at the amalgam/tooth interface. With time in service, this gap fills with corrosion products that have the potential to "seal" the restoration. With the advent of high-copper, more corrosion-resistant amalgams, there has been concern that the time required to create this seal would be increased significantly when compared with low-copper traditional amalgams. The current study was designed to address this concern. Amalgam was condensed into a MACOR mold, simulating a Class I cavity form and then immersed into a 1.0% NaCl solution to simulate oral conditions. Using an air pressure test, the sealing was monitored over time. The results showed that the sealing was influenced by the size of the initial gap prior to immersion as well as corrosion resistance of the amalgam and that a corrosion-resistant amalgam with a small initial gap size can seal as quickly as a corrosion-prone amalgam. Therefore, it is not possible to predict sealing behavior based on corrosion resistance, alone. Furthermore, the presence of zinc in the amalgam alloy has been shown to result in the formation of zinc corrosion products in the amalgam/mold margin, which contributes to more rapid sealing. Analysis of a tooth extracted after 16 years of clinical service that had been restored with an amalgam-containing zinc was also shown to contain zinc corrosion products in the occlusal marginal area. This could explain the reported reduction in marginal fracture of clinically placed amalgam restorations made from zinc containing alloys. PMID- 19544822 TI - Microleakage in resin composite restorations after antimicrobial pre-treatments: effect of KTP laser, chlorhexidine gluconate and Clearfil Protect Bond. AB - The current study evaluated the influence of KTP (Potassium-Titanyl-Phosphate) laser irradiation, 2% chlorhexidine gluconate and Clearfil Protect Bond on the microleakage of Class V composite restorations. Thirty human premolars were selected for cavity pretreatment. After Class V cavity restorations on the buccal and lingual surfaces, the teeth were randomly divided into four groups: Group I Clearfil SE Bond; Group II-KTP laser + Clearfil SE Bond; Group III-2% chlorhexidine gluconate + Clearfil SE Bond; Group IV-Clearfil Protect Bond. The cavities were restored with resin composite. The teeth were then thermocycled for 500 cycles, dried and sealed with nail varnish, leaving 1 mm around the restorations and immersed in 0.5% basic fuchsin for 24 hours. They were then rinsed, dried and sectioned, and microleakage was assessed by dye penetration at the occlusal and gingival surface of the teeth using a stereomicroscope (30x). There were no significant differences among the four groups at the gingival surface (p > 0.05). Microleakage at the occlusal margins of all the groups was compared; differences between the KTP laser and chlorhexidine gluconate group and the KTP laser and Clearfil Protect Bond group were found to be statistically significant (p < 0.05). PMID- 19544823 TI - The influence of luting systems on the microtensile bond strength of dentin to indirect resin-based composite and ceramic restorations. AB - Microtensile bond strength (microTBS) evaluation and fractographic analysis were used to compare four luting systems in the cementation of resin-based composite (RBC) and ceramic disks to dentin. Forty freshly-extracted molars were transversally sectioned to expose flat, deep dentin surfaces. Forty cylindrical specimens (5-mm diameter and 10-mm height), consisting of 20 RBC disks and 20 leucite-based glass ceramic disks, were produced. The RBC disks were sandblasted with 50-microm Al2O3. The ceramic disks were conditioned with 9.5% hydrofluoric acid gel and silane application. All the disks were then bonded to dentin surfaces according to the luting cements to be used: two etch-and-rinse luting agents (XP bond/CoreXFlow; Dentsply [XP]) (Enabond/EnaCem HF; Micerium [ENA]), a self-etch luting system (ED Primer II A+B/Panavia F2.0; Kuraray-Dental [PAN]) and a self-adhesive luting agent (RelyX Unicem; 3M ESPE [UNI]). The adhesive/luting cement systems were applied according to the manufacturers' instructions. The specimens were sectioned perpendicular to the adhesive interface to produce multiple beams, approximately 1 mm2 in area. All the specimen preparations were performed by the same operator. The beams were tested under tension at a crosshead speed of 0.5 mm/minute until failure. The microTBS data were analyzed by two different one-way-ANOVA and multiple comparison Tukey tests (alpha = 0.05). All the fractured beams were observed using a Scanning Electron Microscope (SEM) at 200x magnification for fracture mode determination. The mean bond strength in MPa (SD) for the RBC (Co) and ceramic (Ce) groups were: XP-Co = 31.39 (13.51), ENA-Co = 30.93 (10.17), PAN-Co = 18.29 (10.02), UNI-Co = 19.33 (7.91); XP-Ce = 4.83 (1.86), ENA-Ce = 5.15 (1.66), PAN-Ce = 4.36 (1.80), UNI-Ce = 7.16 (2.52). Statistical analysis showed that the bond strengths were significantly affected by the luting agent employed for both the RBC and ceramic groups (p<0.001). In particular, the XP-Co group and the ENA-Co group did not differ from each other (p > 0.05) and showed significantly higher bond strengths than the PAN-Co and UNI-Co groups (p < 0.05). On the contrary, the UNI-Ce group showed the highest bond strengths compared to the other ceramic experimental groups (p < 0.05). Regarding failure mode, differences were found between the RBC groups: for the etch-and-rinse luting systems (XP-Co and ENA-Co groups), most failures occurred cohesively in the luting agent, while the self-etch luting system (PAN Co group) and self-adhesive luting system (UNI-Co group) failed predominantly adhesively at the luting agent-dentin interface. Little differences were found between the ceramic groups, where failure type was primarily adhesive between cement and ceramic. PMID- 19544824 TI - A comparison of human raters and an intra-oral spectrophotometer. AB - Consistently choosing an accurate shade match is far more difficult than it appears. Recently, several electronic shade-matching devices have been marketed. One device is an intraoral spectrophotometer, Easyshade. The current study compared the accuracy and consistency of the Easyshade (ES) device to three clinicians experienced in tooth whitening trials and trained in the use of the Vitapan 3D Master shade. The maxillary anteriors of 16 participants were matched on three separate occasions one month apart. At each appointment, the three clinicians (R1, R2 & R3) and ES independently chose a single 3D Master tab. A trained research assistant used the Easyshade device to record CIE L*, C* and H* and a shade tab. In addition, color differences between shade tabs were calculated using the Delta E 2000 (delta e 00) formula. The CIE L*C*H* data were also used to establish standards for the five lightness groups of the 3D Master. An intrarater agreement was evaluated using an intraclass correlation statistic, and an inter-rater agreement was evaluated using a weighted Kappa statistic. The percentages of exact matches were: ES = 41%; R1 = 27%; R2 = 22% and R3 = 17%. Matches within a half-shade were also calculated. This represents a mismatch that is perceptible but acceptable. The percentages of matches within a half-tab were: ES = 91%; R1 = 69%; R2 = 85% and R3 = 79%. In terms of lightness, the intra-rater agreement was considered to be very good for ES and R2 and good for R1 and R3. For chroma, agreement for ES was considered good, and for the three clinicians, it was considered moderate. The mean color difference for the L*, C*, H* data recorded at each evaluation was 1.5, or only slightly greater than the color difference between the same tab on different guides (1.2). The delta e 00 data were the most accurate data collected, and they were used to establish a standard to which the tab choices of the four raters were compared. A weighted Kappa statistic was performed and, in terms of lightness, agreement was found to be good for all raters. For chroma, agreement was very good for ES and it was good for the clinicians. In terms of the number of exact matches and matches within a half-shade, the performance of ES was at least comparable to, if not better than, the dentists. Statistically, the same was true in terms of consistency and accuracy when making repeated matches of lightness and chroma using the 3D Master shade guide. PMID- 19544826 TI - Minimally retentive gold onlays: a six-year case report. AB - Partial veneer gold restorations, such as gold onlays, have changed little in preparation design over the past 40 years, as evidenced by textbooks that have long been in print. Such designs must have been based on the assumption that restorations would be luted with zinc phosphate cement but have not altered, despite the introduction of stronger luting cement classes, such as resin composite and resin-modified glass ionomer cements. It is well-established that both porcelain and base metal materials, which have been etched on the intaglio surface, can be combined with resin composite luting cements to produce restorations retained largely through adhesion. However, neither of these materials can equal the combination of minimal tooth reduction and margin adaptation that is possible with high noble gold restorative materials. Although high noble gold alloys cannot be etched, air abrasion of the intaglio surface of restorations likely improves micromechanical retention, but it is unknown whether this, along with a strong luting cement, is sufficient to stabilize high noble gold alloys in preparations with considerably less retention and resistance form than traditional designs. PMID- 19544825 TI - Y-TZP ceramics: key concepts for clinical application. AB - Compared to conventional ceramic systems, Yttrium-stabilized tetragonal zirconia (Y-TZP) ceramics have some superior mechanical properties, ensuring a broad application in dentistry. The current study aimed to present relevant information about Y-TZP ceramics gathered from peer-reviewed papers. A search of the English language peer-reviewed literature was conducted using the PubMed database between 1998 and 2008. Articles that did not focus exclusively on the clinical application of Y-TZP ceramic restorations were excluded from further evaluation. Selected papers describe the chief characteristics of zirconia ceramics and important clinical features, especially those related to cementation procedures. The literature shows that, although new substances and equipment for the surface preparation of zirconia ceramics are in development, the most promising luting protocol seems to be the use of air abrasion with aluminum oxide particles (silanated or not), followed by the application of resin cements or surface primers containing special reactive monomers. However, because zirconia ceramics have only recently been developed for dental applications, there is not enough clinical evidence to support any definitive cementation protocol. PMID- 19544827 TI - A suggested solution for endodontic provisional challenges. PMID- 19544829 TI - Mercury in seals linked to vanishing sea ice. PMID- 19544828 TI - Diagnosis of Sjogren's syndrome from a xerostomia case accompanied by multiple dental caries. PMID- 19544830 TI - In the mix: equine estrogens used in HRT. PMID- 19544831 TI - Calculating the costs of nanohazard testing. PMID- 19544832 TI - Tackling hypoxia in the Baltic Sea: is engineering a solution? PMID- 19544833 TI - Hypoxia-related processes in the Baltic Sea. AB - Hypoxia, a growing worldwide problem, has been intermittently present in the modern Baltic Sea since its formation ca. 8000 cal. yr BP. However, both the spatial extent and intensity of hypoxia have increased with anthropogenic eutrophication due to nutrient inputs. Physical processes, which control stratification and the renewal of oxygen in bottom waters, are important constraints on the formation and maintenance of hypoxia. Climate controlled inflows of saline water from the North Sea through the Danish Straits is a critical controlling factor governing the spatial extent and duration of hypoxia. Hypoxia regulates the biogeochemical cycles of both phosphorus (P) and nitrogen (N) in the water column and sediments. Significant amounts of P are currently released from sediments, an order of magnitude larger than anthropogenic inputs. The Baltic Sea is unique for coastal marine ecosystems experiencing N losses in hypoxic waters below the halocline. Although benthic communities in the Baltic Sea are naturally constrained by salinity gradients, hypoxia has resulted in habitat loss over vast areas and the elimination of benthic fauna, and has severely disrupted benthic food webs. Nutrient load reductions are needed to reduce the extent, severity, and effects of hypoxia. PMID- 19544834 TI - Molecular-level interactions in soils and sediments: the role of aromatic pi systems. AB - This review intends to deepen our understanding of mechanisms by which molecules with aromatic moieties attach to organic and mineral components of terrestrial environments. We present published evidence for the existence of specific, sorptive interactions of aromatic moieties with environmental sorbents. We find that aromatic pi-systems within organic compounds have the capacity to adsorb to minerals and organic soil and sediment components such as natural organic matter (NOM) and fire-derived black carbon (BC) through specific sorptive forces other than hydrophobic interactions. Polar interactions of aromatic pi-donor and acceptor compounds show adsorption energies between 4 and 167 kJ mol(-1). Bonding strengths of cation-pi interactions and pi-pi electron donor-acceptor (EDA) interactions appear to be larger than H bonding strengths and comparable to inner and outer-sphere complex formation. We conclude that, in analogy to polar and ionizable functional groups, components with aromatic pi-donor and -acceptor systems equip organic molecules with a substantial sorptive potential. This observation has important implications for the fate and transport of aromatic contaminants. The resulting sorptive interactions might also play a yet overlooked functional role in the complex chain of processes which preserve NOM against decomposition. PMID- 19544835 TI - Regional nutrient trends in streams and rivers of the United States, 1993-2003. AB - Trends in flow-adjusted concentrations (indicators of anthropogenic changes) and observed concentrations (indicators of natural and anthropogenic changes) of total phosphorus and total nitrogen from 1993 to 2003 were evaluated in the eastern, central, and western United States by adapting the Regional Kendall trend test to account for seasonality and spatial correlation. The only significant regional trend was an increase in flow-adjusted concentrations of total phosphorus in the central United States, which corresponded to increases in phosphorus inputs from fertilizer in the region, particularly west of the Mississippi River. A similar upward regional trend in observed total phosphorus concentrations in the central United States was not found, likely because precipitation and runoff decreased during drought conditions in the region, offsetting the increased source loading on the land surface. A greater number of regional trends would have been significant if spatial correlation had been disregarded, indicating the importance of spatial correlation modifications in regional trend assessments when sites are not spatially independent. PMID- 19544836 TI - Identification of green rust in groundwater. AB - Green rust, a family of Fe(II), Fe(III) layered double hydroxides, is believed to be present in environments close to the Fe(II)/ Fe(III) transition zone. Attempts to identify members of this family in nature have proven difficult because the material is oxidized after only a few minutes exposure to air. In this paper, we present a sampling method for capturing green rust so it is not oxidized. We then we used the method to identify the compound in a groundwater sample taken below the water table from fractures in granite. X-ray diffraction patterns were weak, but clearly identical to those of synthetic GR(CO3), the green rustfamily memberwhere carbonate and water occupy the interlayer between the iron-hydroxide layers. The method was then tested on samples taken from an artesian well and a deep underground experimental station, both within the Fe(II)/ Fe(III) redox zone. In both cases, GR(CO3), could be identified. Currently, transport models for predicting the behavior of contaminants in groundwater do not include parameters for green rust This work demonstrates they should. PMID- 19544837 TI - PCNs, PCDD/Fs, and non-orthoPCBs, in water and bottom sediments from the industrialized Norwegian Grenlandsfjords. AB - Chlorinated toxic planar aromatic compounds were analyzed in the heavily industrialized Grenlandsfjords, which is a system of silled fjords in southern Norway. Surface water samples contained 7.4-160 ng/m3 polychlorinated dibenzo-p dioxins and furans (PCDD/Fs), 14-410 ng/m3 polychlorinated naphthalenes (PCNs), and 0.31-2.4 ng/m3 non-orthochlorinated biphenyls (non-orthoPCBs). The concentrations of PCDD/Fs were about 300 times higher than in the Baltic Sea. Highest level of the compounds wasfound near a magnesium production plant Hepta CDFs and penta-CNs dominated in the inner-fjord waters, and tetra-CB 77 was the major non-orthoPCB congener. Sediment samples had PCDD/F concentration of 25-730 ng/g dw. Highest concentration was detected close to the magnesium plant. Octa CDFdominated in the fjord sediments, especially near the magnesium plant indicating a discharge-specific contamination with this congener. The isomer composition of PCDD/Fs and of PCNs, was unchanged when comparing samples from different layers of a sediment core from the deep anoxic water. This concludes that essentially zero degradation had occurred during approximately 50 years in this environment. PMID- 19544838 TI - A comparison of summertime secondary organic aerosol source contributions at contrasting urban locations. AB - Primary and secondary sources contributing to atmospheric organic aerosol during the months of July and August were quantitatively assessed in three North American urban areas: Cleveland, Ohio, and Detroit, Michigan, in the Midwest region and Riverside, California, in the Los Angeles Air Basin. Organic molecular marker species unique to primary aerosol sources and secondarytracers derived from isoprene, alpha-pinene, beta-caryophyllene, and toluene were measured using gas chromatography-mass spectrometry. Source contributions from motor vehicles, biomass burning, vegetative detritus, and secondary organic aerosol (SOA) were estimated using chemical mass balance (CMB) modeling. In Cleveland, primary sources accounted for 37 +/- 2% of ambient organic carbon, measured biogenic and anthropogenic secondary sources contributed 46 +/- 6%, and other unknown sources contributed 17 +/- 4%. Similarly, Detroit aerosol was determined to be 44 +/- 5% primary and 37 +/- 3% secondary, while 19 +/- 7% was unaccounted for by measured sources. In Riverside, 21 +/- 3% of organic carbon came from primary sources, 26 +/- 5% was attributed to measured secondary sources, and 53 +/- 3% came from other sources that were expected to be secondary in nature. The comparison of samples across these two regions demonstrated that summertime SOA in the Midwestern United States was substantially different from the summertime SOA in the Los Angeles Air Basin and indicated the need to exert caution when generalizing about the sources and nature of SOA across different urban areas. Furthermore, the results of this study suggestthatthe contemporary understanding of SOA sources and formation mechanisms is satisfactory to explainthe majority of SOA in the Midwest Additional SOA sources and mechanisms of formation are needed to explain the majority of SOA in the Los Angeles Air Basin. PMID- 19544839 TI - Tetracycline resistance genes and tetracycline resistant lactose-fermenting Enterobacteriaceae in activated sludge of sewage treatment plants. AB - Activated sludges were sampled from five sewage treatment plants (STPs) distributed in three geographically isolated areas, i.e., Hong Kong (Shatin, Stanley), Shanghai (Minhang) in China, and the bay area in California (Palo Alto and San Jose) of the United States. Among the tested 14 tetracycline resistance (tet) genes, nine genes encompassing efflux pumps (tetA, tetC, tetE, and tetG), ribosomal protection proteins (tetM, tetO, tetQ, and tetS), and enzymatic modification (tetX) were commonly detected in the STP sludge samples, whereas five genes encompassing efflux pumps [tetB, tetD, tetL, tetK, and tetA(P)] were not detected in any sludge sample. Additionally, 109 lactose-fermenting Enterobacteriaceae (LFE) strains were isolated from the activated sludge of the Shatin STP. Tetracycline-resistant (TR) LFE accounted for 32% of the total 109 LFE strains. The occurrence frequencies of tet genes among all TR-LEF strains varied from 0 to 91%, i.e., tetC (91%), tetA (46%), tetE (9%), tetG (6%), and tetD (6%). Finally, quantitative real-time polymerase chain reaction was used to quantify the change of tetC and tetA genes as the indicator of TR-LEF in the Shatin and Stanley STPs. The results showed that the concentrations of tetC and tetA genes in STP effluent ranged from 10(4) to 10(5) copies/mL, significantly lower than those in the influent by 3 orders of magnitude. PMID- 19544840 TI - Effect of climate change between 1984 and 2007 on precipitation chemistry at a site in northeastern U.S.A. AB - Climate change predictions for the northeastern US call for an increase in tropical storms and a decrease in extra tropical cyclones including continental storms. We ran 24-h back trajectories for each precipitation event that occurred at the Cary Institute of Ecosystem Studies in southeastern New York, U.S.A. from 1984 to 2007 and analyzed precipitation chemistry as well as air mass position 24 h prior to the onset of each precipitation event. The results showed an increase in marine precipitation and a slight but statistically insignificant decrease in continental precipitation during the 1984-2007 period. The chemistry of precipitation from the two directions was quite different marine storms were higher in Na4 and Cl- but lower in solutes associated with acid precipitation (H+, SO4(2-), NO3-, and NH4+). Annual mean concentrations of acid precipitation solutes declined for storms from both directions during the period. We used a simple mixing model based on the current rates of increase and decrease of marine and continental precipitation respectively to show that chemical changes in precipitation resulting from the shift in storm tracks are small compared to chemical changes due to emissions reductions. PMID- 19544841 TI - PAH biodegradative genotypes in Lake Erie sediments: evidence for broad geographical distribution of pyrene-degrading mycobacteria. AB - Despite a long history of anthropogenic contamination of Lake Erie sediments, little work has been done to understand the potential for PAH biodegradation by indigenous microbial communities. Pyrene-degrading Mycobacterium are prevalent in many polycyclic aromatic hydrocarbon (PAH)-contaminated freshwater sediments, and are of interest for their ability to degrade environmentally recalcitrant high molecular weight PAHs. This work tested the hypothesis that pyrene-degrading mycobacteria are prevalent in Lake Erie; an additional aim was to gain a baseline picture of the sediment microbial communities through sequencing a 16S rDNA clone library. Biodegradation potential of Lake Erie Mycobacterium populations was assessed through quantification of pyrene dioxygenase genes (nidA) and mycobacteria 16S rDNA genes using quantitative real time PCR. nidA was detected at all seven sampling sites across Lake Erie, with abundances ranging from 2.09 to 70.4 x 10(6) copies per gram sediment, with highest abundances at the most PAH contaminated site (Cleveland Harbor). This is in contrastto naphthalene dioxygenase genes commonly used as biomarkers of PAH degradation: nahAc (from gamma-proteobacteria) was not detected anywhere, and nagAc (from beta proteobacteria) was detected only in Cleveland Harbor, despite dominance by proteobacteria in Lake Erie sediment 16S rDNA clone libraries (>50% of clones). The prevalence of Mycobacterium nidA genotypes corroborated previous studies indicating that PAH-degrading mycobacteria have a cosmopolitan distribution and suggests they play an important but overlooked role in natural attenuation and cycling of PAHs in Lake Erie. PMID- 19544842 TI - Exploring estuarine nutrient susceptibility. AB - The susceptibility of estuaries to nutrient loading is an important issue that cuts across a range of management needs. We used a theory-driven but data-tested simple model to assist classifying estuaries according to their susceptibility to nutrients. This simple nutrient-driven phytoplankton model is based on fundamental principles of mass balance and empirical response functions for a wide variety of estuaries in the United States. Phytoplankton production was assumed to be stoichiometrically proportional to nitrogen load and an introduced "efficiency factor" intended to capture the myriad processes involved in converting nitrogen load to algal production. A Markov Chain Monte Carlo algorithm of Bayesian inference was then employed for parameter estimation. The model performed remarkably well for chlorophyll estimates, and the predicted estimates of primary production, grazing, and sinking losses are consistent with measurements reported in the literature from a wide array of systems. Analysis of the efficiency factor suggests that estuaries with the ratio of river inflow to estuarine volume (Q/V) greater than 2.0 per year are less susceptible to nutrient loads, and those with Q/V between 0.3 and 2.0 per year are moderately susceptible. This simple model analysis provides a first-order screening tool for estuarine susceptibility classification. PMID- 19544843 TI - Groundwater pollution by perfluorinated surfactants in Tokyo. AB - Perfluorinated surfactants (PFSs) in groundwater were analyzed to reveal their distribution and sources. Sixteen groundwater and spring samples were collected from the Tokyo metropolitan area, and nine PFSs, including perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA), were analyzed by liquid chromatography-tandem mass spectrometry. A column test using artificial street runoff was also performed to study their behavior. PFSs were detected in all groundwater samples, some at concentrations comparable to those in wastewater and street runoff, suggesting widespread contamination of groundwater by PFSs. In particular, PFOS -was more abundant in groundwater than in rivers, wastewater, and street runoff. This was attributed to its production from the degradation of its precursors, as supported by the column test. The occurrence of short-chain perfluorocarboxylates (PFCAs) in groundwater was also consistent with the results of the column test, showing that limited amounts of short-chain PFCAs were removed by soil, as the efficiency of removal increased with the chain length. We evaluated the contributions of PFCAs from wastewater and surface runoff to groundwater by using two indicators, the long/(short + long) ratio and the even(even + odd) ratio. Both ratios showed good agreement in their calculated contributions in heavily contaminated groundwater where breakthroughs likely occurred. Wastewater and surface runoff contributed to 54-86% and 16-46% of PFCAs, respectively, in groundwater. PMID- 19544844 TI - Linking the diffusion of water in compacted clays at two different time scales: tracer through-diffusion and quasielastic neutron scattering. AB - Diffusion of water and solutes through compacted clays or claystones is important when assessing the barrier function of engineered or geological barriers in waste disposal. The shape and the connectivity of the pore network as well as electrostatic interactions between the diffusant and the charged clay surfaces or cations compensating negative surface charges affect the resistance of the porous medium to diffusion. Comparing diffusion measurements performed at different spatial or time scales allows identification and extraction of the different factors. We quantified the electrostatic constraint q for five different highly compacted clays (rhob = 1.85 +/- 0.05 g/cm3) using quasielastic neutron scattering (QENS) data. We then compared the QENS data with macroscopic diffusion data for the same clays and could derive the true geometric tortuosities G of the samples. Knowing the geometric and electrostatic factors for the different clays is essential when trying to predict diffusion coefficients for other conditions. We furthermore compared the activation energies Ea for diffusion at the two measurement scales. Because Ea is mostly influenced by the local, pore scale surroundings of the water, we expected the results to be similar at both scales. This was indeed the case for the nonswelling clays kaolinite and illite, which had Ea values lower than that of bulk water, but not for montmorillonite, which had values lower than that in bulk water at the microscopic scale, but larger at the macroscopic scale. The differences could be connected to the strongly temperature dependent mobility of the cations in the clays, which may act as local barriers in the narrow pores at low temperatures. PMID- 19544845 TI - Polybrominated diphenyl ethers and their hydroxylated analogues in ringed seals (Phoca hispida) from Svalbard and the Baltic Sea. AB - The present study investigated the concentrations and patterns of PBDEs and hydroxylated (OH) PBDE analogues in two ringed seal populations: less contaminated Svalbard and more contaminated Baltic Sea. Mean concentration of hepatic sigma-PBDE, which was dominated by BDE47, was six times higher in the ringed seals from the Baltic Sea compared to the seals from Svalbard. BDE47/sigma PBDE was higher in the seals from Svalbard compared to that for Baltic seals, while the trend was opposite for BDE153 and 154. The geographical difference in contaminant pattern of PBDEs in ringed seals could be explained by biotransformation via oxidative metabolism and/or by dietary differences. OH PBDEs were detectable in the majority of plasma samples from both locations, and dominated by bioaccumulation of naturally occurring congeners. Low levels of 3-OH BDE47 and 4'-OH-BDE49 in the Baltic ringed seals suggested minor oxidative biotransformation of BDE47. In the Baltic seals, BDE153/sigma-PBDEs and BDE154/sigma-PBDEs increased and BDE28/sigma-PBDE decreased with increasing sigma POP concentration, which suggests BDE153 and 154 are more persistent than BDE28. Contrasting diets of the ringed seals in these two locations may influence the PBDE congener pattern due to selective long-range transport and direct effluent emissions to Svalbard and the Baltic, respectively. PMID- 19544846 TI - Impact of emissions from the Los Angeles port region on San Diego air quality during regional transport events. AB - Oceangoing ships emit an estimated 1.2-1.6 million metric tons (Tg) of PM10 per year and represent a significant source of air pollution to coastal communities. As shown herein, ship and other emissions near the Los Angeles and Long Beach Port region strongly influence air pollution levels in the San Diego area. During time periods with regional transport, atmospheric aerosol measurements in La Jolla, California show an increase in 0.5-1 microm sized single particles with unique signatures including soot, metals (i.e., vanadium, iron, and nickel), sulfate, and nitrate. These particles are attributed to primary emissions from residual oil sourcessuch as ships and refineries, as well as traffic in the port region, and secondary processing during transport. During regional transport events, particulate matter concentrations were 2-4 times higher than typical average concentrations from local sources, indicating the health, environmental, and climate impacts from these emission sources must be taken into consideration in the San Diego region. Unless significant regulations are imposed on shipping related activities, these emission sources will become even more important to California air quality as cars and truck emissions undergo further regulations and residual oil sources such as shipping continue to expand. PMID- 19544847 TI - Initial estimates of mercury emissions to the atmosphere from global biomass burning. AB - The average global annual mercury emission estimate from biomass burning (BMB) for 1997-2006 is 675 +/- 240 Mg/year. This is equivalentto 8% of all currently known anthropogenic and natural mercury emissions. By season, the largest global emissions occur in August and September, the lowest during northern winters. The interannual variability is large and region-specific, and responds to drought conditions. During this particular time period, the largest mercury emissions are from tropical and boreal Asia, followed by Africa and South America. They do not coincide with the largest carbon biomass burning emissions, which originate from Africa. Frequently burning grasslands in Africa and Australia, and agricultural waste burning globally, contribute relatively little to the mercury budget The released mercury from BMB is eventually deposited locally and globally and contributes to the formation of toxic bioaccumulating methyl mercury. Furthermore, increasing temperature in boreal regions, where the largest soil mercury pools reside, is expected to exacerbate mercury emission because of more frequent larger, and more intense fires. PMID- 19544848 TI - Mercury in a boreal forest stream--role of historical mercury pollution, TOC, temperature, and water discharge. AB - Over a one-year study period (2003), we monitored total Hg (HgT) and methyl Hg (MeHg) at two sites in a Swedish forest stream located above (Site(ref)) and below a stretch of Hg-contaminated sediments (SiteHg). We also monitored HgT, MeHg, and ancillary water chemistry in peat water close to the stream and HgT in open field wet deposition. Despite the presence of historical Hg contaminants, direct atmospheric Hg deposition and transfer of Hg from the catchment explained more than half of the annual HgT load at SiteHg. The concentrations of both HgT and MeHg were sensitive to changes in water discharge (Q) and water temperature (T) at both sites, suggesting that the stream HgT and MeHg load can change dramatically in response to changing weather conditions. The 2003 data together with data from 1996 disclosed intersite differences and temporal variation in the relationships between HgT, MeHg, and TOC (total organic carbon), reflecting variable sources of HgT, MeHg, and TOC and temporal changes in factors affecting Hg speciation. PMID- 19544849 TI - Two-decade reconstruction of algal blooms in China's Lake Taihu. AB - The algal blooming in the inland lakes has become a critically important issue for its impacts not only on local natural and social environments, but also on global human community. However, the occurrences of blooming on larger spatial scale and longer time scale have rarely been studied. As the third largest freshwater lake in China, Lake Taihu has drawn increasing attention from both public and scientific communities concerning its degradation. Using available satellite images, we reconstructed the spatial and temporal patterns of algal blooms in Lake Taihu through the pasttwo decades. The blooming characteristics over the past two decades were examined with the dynamic of initial blooming date being highlighted. The initial blooming dates were gradually becoming later and later from 1987 to 1997. Since 1998, however, the initial blooming date came earlier and earlier year by year, with approximately 11.42 days advancement per year. From 1987 to 2007, the annual duration of algal blooms lengthened year by year, in line with the substantial increases in the occurrences of algal blooms in spring and summer months. The algal blooms usually occur in northern bays and spread to center and south parts of Lake Taihu. The increases in previous winter's mean daily minimum temperature partially contributed to the earlier blooming onset. However, human activities, expressed as total gross domestic product (GDP) and population, outweighed the climatic contribution on the initial blooming date and blooming duration. This study may provide insights for the policy makers who try to curb the algal blooming and improve the water quality of inland freshwater lakes. PMID- 19544850 TI - Functional gene array-based analysis of microbial community structure in groundwaters with a gradient of contaminant levels. AB - To understand how contaminants affect microbial community diversity, heterogeneity, and functional structure, six groundwater monitoring wells from the Field Research Center of the U.S. Department of Energy Environmental Remediation Science Program (ERSP; Oak Ridge, TN), with a wide range of pH, titrate, and heavy metal contamination were investigated. DNA from the groundwater community was analyzed with a functional gene array containing 2006 probes to detect genes involved in metal resistance, sulfate reduction, organic contaminant degradation, and carbon and nitrogen cycling. Microbial diversity decreased in relation to the contamination levels of the wells. Highly contaminated wells had lower gene diversity but greater signal intensity than the pristine well. The microbial composition was heterogeneous, with 17-70% overlap between differentwells. Metal-resistant and metal-reducing microorganisms were detected in both contaminated and pristine wells, suggesting the potential for successful bioremediation of metal-contaminated groundwaters. In addition, results of Mantel tests and canonical correspondence analysis indicate that nitrate, sulfate, pH, uranium, and technetium have a significant (p < 0.05) effect on microbial community structure. This study provides an overall picture of microbial community structure in contaminated environments with functional gene arrays by showing that diversity and heterogeneity can vary greatly in relation to contamination. PMID- 19544851 TI - Factors influencing concentrations of polybrominated diphenyl ethers (PBDEs) in students from Antwerp, Belgium. AB - Human exposure to polybrominated diphenyl ethers (PBDEs) through food and indoor dust ingestion was assessed for 19 Belgian adults. The intake of PBDEs (sigma hepta BDEs and BDE 209) in the studied population is influenced mainly by diet Dietary intakes of sigmatri-hepta BDEs (BDEs 28, 47, 99, 100, 153, 154, and 183) were 5.9-22.0 ng/day (median 10.3), while those via dust ingestion were 0.1-1.4 ng/day (median 0.25) or 0.3-3.5 ng/day (average 0.6), assuming dust ingestion rates of 20 and 50 mg/day, respectively. Dietary intakes of BDE 209 were 50-238 ng/day (median 95), whereasthose via dust ingestion were 0.4-11 ng/day (median 1.8) or 1.0-29 ng/day (median 4.6) for dust ingestion rates of 20 and 50 mg/day, respectively. It is important to acknowledge the uncertainty associated with the dust ingestion rates. Concentrations of sigmatri-hepta BDEs measured in blood serum were 0.9-7.2 ng/g lipid weight (lw) (median 1.9). This is similar to other European populations, but lower than for nonoccupationally exposed Americans (average of 19 ng/g lw). When compared with estimates of exposure via both dietary and indoor dust ingestion for Americans, the exposures reported here are consistent with the hypothesis that the difference between European and American body burdens of PBDEs is attributable primarily to greater exposure via dust ingestion for Americans. The total intake of PBDEs through food and dustfor each participant could not be correlated with the corresponding serum concentration. Instead, it is hypothesized that past and episodic current higher intakes of PBDEs are more important determinants of body burden than continuous background exposures at the low levels measured in this study. PMID- 19544852 TI - Weathering and the fallout plume of heavy oil from strong petroleum seeps near Coal Oil Point, CA. AB - The Coal Oil Point (COP) seeps offshore Goleta, CA, are estimated to release 20 25 tons of oil daily, providing an ideal natural laboratory to investigate the fate of oil in the coastal ocean. To address the long-term fate of COP oil, we collected 15 sediment samples down current from the seeps and quantified petroleum content and individual biomarkers using traditional and comprehensive two-dimensional gas chromatography. Similarities in the distributions of hopane biomarkers link the oil in the sediments to fresh seep oil (n=5) and underlying reservoirs (n=3), although sediment oil is heavily weathered. The spatial distribution of oil forms a plume along the continental shelf that we suggest represents a chronic fallout pattern for heavy oil from the persistent surface slicks; average surface currents appear to modulate the distribution of the fallout over a period of 0.4-5 days. The extent of hydrocarbon loss is consistent for all sediments, indicating a common limit to oil weathering with contributions from evaporation, biodegradation, and dissolution. Considering the amount of oil and quantity of sediment impacted, we estimate a sediment oil burden of 0.3 x 10(12) to 3 x 10(12) g in the study area, equivalent to 8-80 spills of the Exxon Valdez accident of 1989. PMID- 19544853 TI - Characterization of metabolites formed during the biotransformation of 17alpha ethinylestradiol by Nitrosomonas europaea in batch and continuous flow bioreactors. AB - The biotransformation of 17alpha-ethinylestradiol (EE2) by an ammonia oxidizing bacteria, Nitrosomonas europaea, grown in batch (ammonia-rich) and continuous flow (chemostat, ammonia-limited) reactors was investigated. Both C-14 labeled EE2 (10 gammag/L) and unlabeled EE2 (1 mg/L) were used to facilitate metabolite identification under environmentally relevant physiological conditions. Whole cell ammonia monooxygenase (AMO) activity was not inhibited at the EE2 concentrations used in this study. Characterization of the primary metabolite formed during batch cultivation by liquid chromatography/ion-trap mass spectrometry (LC-ITMS) and nuclear magnetic resonance (NMR) spectroscopy showed modification at the ethinyl group and addition of a carboxyl group. This metabolite (M386) (revealed by m/z 385 in negative mode electrospray LC/ MS) was not formed in the abiotic control. In contrast, biotransformation of EE2 under continuous flow conditions showed formation of a monohydroxylated EE2 (revealed by m/z 311), but not M386. Furthermore, nitrated EE2 derivatives were formed in both batch and continuous flow cultures, as a result of abiotic transformation of EE2 in the presence of high concentrations of nitrite in the bioreactors. Results from this study underscore the importance of physiological state and growth conditions as critical variables that can dictate the metabolic pathway for EE2 biodegradation and the nature of byproducts formed. PMID- 19544854 TI - Uptake and acropetal translocation of polycyclic aromatic hydrocarbons by wheat (Triticum aestivum L.) grown in field-contaminated soil. AB - Uptake and acropetal translocation of 14 priority polycyclic aromatic hydrocarbons (PAHs) by wheat (Triticum aestivum L.) grown in 15 field contaminated soils were investigated in a growth chamber. PAH concentrations in roots correlated positively with the corresponding concentrations in soils and negatively with the contents of soil organic carbon (p < 0.01). No clear linear relationship was found between log RCF (root concentration factor, microg g(-1) root/microg g(-1) soil on dry weight basis) and log Kow of these PAHs. Four-ring PAHs had the highest tendency to be taken up by roots. PAH concentrations in shoots correlated well with their concentrations in soils and roots. Furthermore, distribution profiles of PAHs in shoots were fairly similar to those in soils. Acropetal translocation of 10 PAHs (with log Kow varying from 3.45 to 5.78) was also implicated by Rt (ratio of PAH from root-to-shoot translocation to the total accumulation in shoots) ranging from 53.6 to 72.6%. A negative linear relationship was found between log Rt and log Kow of these PAHs (p < 0.01), and acropetal translocation of PAHs depended on their chemical properties. PMID- 19544855 TI - Yields of carbonyl products from gas-phase reactions of fragrance compounds with OH radical and ozone. AB - Chamber studies to quantify formation yields of oxygenated organic reaction products were performed for gas-phase reactions of the hydroxyl radical (OH*) and ozone (03) with the common cleaning product terpene compounds limonene, alpha terpineol, and geraniol. The reaction products observed were identified and quantified using derivatization by O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) and gas chromatography/mass spectrometry. Limonene rate constants and product mechanisms have been examined previously. Several of these investigations have measured productyields from limonene reactions and those results are compared with the results presented here. Although rate constants and product mechanisms have previously been investigated for alpha-terpineol and geraniol, yields of oxygenated organic reaction products have not been measured. Reactions from the fragrance compounds in this study produced several dicarbonyl reaction products such as glyoxal, methylglyoxal, and 4-oxopentanal which were observed from all three terpenes. Total carbonyl yields ranged from 5.1% for the limonene + O3 reaction to 92% for the geraniol + O3 reaction. PMID- 19544856 TI - Temperature sensitivity indicates that chlorination of organic matter in forest soil is primarily biotic. AB - Old assumptions that chloride is inert and that most chlorinated organic matter in soils is anthropogenic have been challenged by findings of naturally formed organochlorines. Such natural chlorination has been recognized for several decades, but there are still very few measurements of chlorination rates or estimates of the quantitative importance of terrestrial chlorine transformations. While much is known about the formation of specific compounds, bulk chlorination remains poorly understood in terms of mechanisms and effects of environmental factors. We quantified bulk chlorination rates in coniferous forest soil using 36Cl-chloride in tracer experiments at different temperatures and with and without molecular oxygen (O2). Chlorination was enhanced by the presence of O2 and had a temperature optimum at 20 degrees C. Minimum rates were found at high temperatures (50 degrees C) or under anoxic conditions. The results indicate (1) that most of the chlorination between 4 and 40 degrees C was biotic and driven by O2 dependent enzymes, and (2) that there is also slower background chlorination occurring under anoxic conditions at 20 degrees C and under oxic conditions at 50 degrees C. Hence, while oxic and biotic chlorination clearly dominated, chlorination by other processes including possible abiotic reactions was also detected. PMID- 19544857 TI - Kinetics of C60 fullerene dispersion in water enhanced by natural organic matter and sunlight. AB - The industrial-scale production of Buckminster fullerene C60 elicits concerns over its impact on human health and ecosystems because of the reported, albeit debatable, toxicity. Assessment of the overall environment risk requires a good estimate of the level of exposure and careful characterization of the physicochemical properties of C60 in natural aqueous environments. The reported study investigates the role of various environmental factors, i.e., ionic composition, natural organic matter (NOM), and light in dispersion of C60 in the aqueous phase by simple mixing. The presence of NOM greatly enhances C60 dispersion, and the dispersion process is further accelerated by sunlight. At typical NOM concentrations found in natural waters, C60 concentrations of a few to tens of milligrams per liter can occur within 10 days of mixing, regardless of its extremely low water solubility. The rate of dispersing decreases with the increase of ionic strength. However, calcium ions significantly increase C60 concentration in the aqueous phase. Results from UV/vis absorbance characterization strongly suggest that C60 may have been chemically modified when dispersed in an NOM solution in the presence of sunlight. This reaction pathway has significant implication on the fate, transport, and environmental impact of C60 fullerene. PMID- 19544858 TI - Role of specific ion interactions in seawater RO membrane fouling by alginic acid. AB - Organic fouling plagues many environmental membrane processes. In this study, well-controlled laboratory experiments were performed to elucidate seawater RO membrane fouling by alginic acid. Interfacial free energies derived from multiple probe liquid contact angle analyses (including different seawater matrices) correlated strongly with the rates of membrane fouling. More importantly, the Lewis acid-base interfacial free energy quantitatively described the impacts of calcium-carboxylate complex formation and predicted membrane fouling and cleaning behavior. Calcium ions made polyamide composite RO membranes (and alginic acid) more hydrophobic, enhanced the rate and extent of flux decline, and reduced the effectiveness of chemical cleaning. The implications for seawater RO membrane fouling are clear. Selective removal of calcium ions via pretreatment can reduce the gel forming ability of carboxylate rich biomacromolecules and, hence, the extent to which they foul RO membranes. In addition, RO membranes should be produced with smooth, hydrophilic interfaces comprising monopolar electron-donor functionality and no carboxylic acid residue. More broadly, this paper presents a facile approach for quantifying the impacts of specific ion interactions on aquatic colloid stability, aggregation, and deposition. PMID- 19544859 TI - Soil burdens of persistent organic pollutants: their levels, fate, and risks. Part iv. Quantification of volatilization fluxes of organochlorine pesticides and polychlorinated biphenyls from contaminated soil surfaces. AB - A volatilization chamber, designed for direct measurements of the soil-air exchange of persistent organic pollutants (POPs) was applied for determination of the volatilization fluxes of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs). The volatilization fluxes were determined for 13 model compounds at 3-5 concentration levels, for two soil organic carbon contents, and two wind velocities. The flux values were strongly correlated with physicochemical properties of the compounds. The higher fluxes were measured for soils with lower organic carbon contents, for higher contamination, and higher wind velocities. Experimentally derived values were compared to those predicted by the fugacity model. In general, the fugacity model underestimated the volatilization fluxes, especially for the compounds with higher molecular weights, and soils with higher organic carbon contents. It has been demonstrated that variability of the wind velocities as an important parameter for quantification of the soil-air exchange should be better considered in current models. Presented results draw the attention to often overlooked secondary sources of the atmospheric pollution and point out that their impact can be much greater than indicated by the fugacity models. PMID- 19544860 TI - PbO2(s, plattnerite) reductive dissolution by aqueous manganous and ferrous ions. AB - Pb(IV)O2(s, plattnerite) nanoparticle aggregrates in aqueous suspension are readily reduced by Mn2+(aq) and Fe2+(aq) between pH 3.0 and 8.5, yielding pb2+(aq) and adsorbed Pb". Fe2+(aq) oxidation generates Fe(III) (hydr)oxides that impede Pb(IV) reduction, especially at pH > or =5. Under acidic conditions, production of dissolved Fe(III) may also be significant. Mn2+(aq) oxidation generates mixed Mn(III)/Mn(IV) (hydr)oxides that are less of an impediment to Pb(IV) reduction. Adding both Fe2+(aq) and Mn2+(aq) can set in motion a Mn redox cycle that catalyzes PbO2(s) reduction by Fe2+(aq). Reaction with Fe2+(aq) and Mn2+(aq) can affect subsequent reactions with natural organic matter. Hydroquinone was employed as a representative organic reductant. Both hydroquinone and its two-electron oxidation product p-benzoquinone are readily quantified by HPLC. Adding Fe2+(aq) before hydroquinone greatly diminishes p benzoquinone production. Adding Mn2+(aq) before hydroquinone has little effect on p-benzoquinone production. Free chlorine residual variations in premise plumbing can setthe stage for the reactions documented here. PMID- 19544861 TI - PbO2(s, plattnerite) reductive dissolution by natural organic matter: reductant and inhibitory subfractions. AB - Natural organic matter (NOM) is a diverse collection of molecules, each possessing its own reductant, complexant, and adsorption properties. Here, we are interested in the ability of NOM to bring about the reductive dissolution of Pb(IV)O2(s). Adding the coagulants FeCl3 or Al2(SO4)3 followed by membrane filtration is one way to remove a subset of NOM molecules from surface water samples. Another is to pass water samples through a granular activated carbon (GAC) column. Results from applying these treatments to Great Dismal Swamp water (DSW) and Nequasset Bog Water (NBW) can best be explained as follows: (i) GAC column treatment is more efficient at removing the NOM fraction most responsible for reductive dissolution. (ii) Coagulation/filtration, with either coagulant, is most efficient at removing a second, inhibitory fraction. Inhibition may arise from (i) adsorption at the mineral/water interface, which blocks approach of reductant molecules and (ii) a micelle-like aggregate nature, which provides hydrophobic pockets that capture reductantmolecules, again keeping them away from the mineral/water interface. Hypotheses regarding reductant and inhibitory fractions are further evaluated using representative low-molecular-weight compounds. Substituted hydroquinones are used as mimics of the reductant fraction, and malonic acid, quinic acid, trehalose, alginic acid, and polygalacturonic acid are used as mimics of the inhibitory fraction. PMID- 19544862 TI - Fate of conjugated natural and synthetic steroid estrogens in crude sewage and activated sludge batch studies. AB - Steroids are excreted from the human body in the conjugated form but are present in sewage influent and effluent as the free steroid, the major source of estrogenic activity observed in water courses. The fate of sulfate and glucuronide conjugated steroid estrogens was investigated in batch studies using activated sludge grown on synthetic sewage in a laboratory-scale Husmann simulation and crude sewage from the field. A clear distinction between the fate of sulfate and glucuronide conjugates was observed in both matrices, with sulfated conjugates proving more recalcitrant and glucuronide deconjugation preferential in crude sewage. For each conjugate, the free steroid was observed in the biotic samples. The degree of free steroid formation was dependent on the conjugate moiety, favoring the glucuronide. Subsequent degradation of the free steroid (and sorption to the activated sludge solid phase) was evaluated. Deconjugation followed the first order reaction rate with rate constants for 17alpha-ethinylestradiol 3-glucuronide, estriol l6alpha-glucuronide, and estrone 3-glucuronide determined as 0.32, 0.24, and 0.35 h respectively. The activated sludge solid retention time over the range of 3-9 days had 74 to 94% of sulfate conjugates remaining after 8 h. In contrast, a correlation between increasing temperature and decreasing 17alpha-ethinylestradiol 3-glucuronide concentrations in the activated sludge observed no conjugate present in the AS following 8 h at 22 degrees C Based on these batch studies and literature excretion profiles, a hypothesis is presented on which steroids and what form (glucuronide, sulfate, or free) will likely enter the sewage treatment plant. PMID- 19544863 TI - Measurement and estimation of organic-liquid/water interfacial areas for several natural porous media. AB - The objective of this study was to quantitatively characterize the impact of porous-medium texture on interfacial area between immiscible organic liquid and water residing within natural porous media. Synchrotron X-ray microtomography was used to obtain high-resolution, three-dimensional images of solid and liquid phases in packed columns. The image data were processed to generate quantitative measurements of organic-liquid/water interfacial area and of organic-liquid blob sizes. Ten porous media, comprising a range of median grain sizes, grain-size distributions, and geochemical properties, were used to evaluate the impact of porous-medium texture on interfacial area. The results show that fluid-normalized specific interfacial area (A) and maximum specific interfacial area (Am) correlate very well to inverse median grain diameter. These functionalities were shown to result from a linear relationship between effective organic-liquid blob diameter and median grain diameter. These results provide the basis for a simple method for estimating specific organic-liquid/water interfacial area as a function of fluid saturation for a given porous medium. The availability of a method for which the only parameter needed is the simple-to-measure median grain diameter should be of great utility for a variety of applications. PMID- 19544864 TI - Distribution of proton dissociation constants for model humic and fulvic acid molecules. AB - The intrinsic proton binding constants of 10 model humic acid and six model fulvic acid molecules are calculated using SPARC Performs Automated Reasoning in Chemistry (SPARC). The accuracy of the SPARC calculations is examined using estimated microscopic binding constants of various small organic acids. An equimolar mixture of the appropriate hypothetical molecules is used as a representation of soil and aqueous humic acid and fulvic acid. The probability distributions of the mixture microscopic proton binding constants and the intrinsic proton binding constants in the metal speciation models WHAM V and WHAM VI (Windermere humic aqueous models) are compared. The idea is to assess the predictive value of the molecular mixture models as representations of heterogeneous natural organic matter. For aqueous humic and fulvic acids, the results are comparable to the WHAM distribution. For soil humic acid, the WHAM probability distribution is less acidic for the carboxylic sites but similar to that of the phenolic sites. Computations made using the WHAM molecular distributions and WHAM VI are comparable to titration data for Suwannee River fulvic acid. These results suggest that mixture molecular models can be used to investigate and predict the binding of metal cations to humic and fulvic acids. PMID- 19544865 TI - Stormwater sediment and bioturbation influences on hydraulic functioning, biogeochemical processes, and pollutant dynamics in laboratory infiltration systems. AB - Stormwater sediments that accumulate at the surface of infiltration basins reduce infiltration efficiencies by physical clogging and produce anoxification in the subsurface. The present study aimed to quantify the influence of stormwater sediment origin (urban vs industrial catchments) and the occurrence of bioturbators (tubificid worms) on the hydraulic functioning, aerobic/anaerobic processes, and pollutant dynamics in stormwater infiltration systems. In laboratory sediment columns, effects of stormwater sediments and tubificids were examined on hydraulic conductivity, microbial processes, and pollutant releases. Significant differences in physical (particle size distribution) and chemical characteristics betoveen the two stormwater sediments led to distinct effects of these sediments on hydraulic and biogeochemical processes. Bioturbation by tubificid worms could increase the hydraulic conductivity in stormwater infiltration columns, but this effect depended on the characteristics of the stormwater sediments. Bioturbation-driven increases in hydraulic conductivity stimulated aerobic microbial processes and enhanced vertical fluxes of pollutants in the sediment layer. Our results showed that control of hydraulic functioning by stormwater sediment characteristics and/ or biological activities (such as bioturbation) determined the dynamics of organic matter and pollutants in stormwater infiltration devices. PMID- 19544866 TI - Arsenic redox changes by microbially and chemically formed semiquinone radicals and hydroquinones in a humic substance model quinone. AB - Arsenic is a redox-active metalloid whose toxicity and mobility strongly depends on its oxidation state, with arsenite (As(III)) being more toxic and mobile than arsenate (As(V)). Humic substances (HS) are also redox-active and can potentially react with arsenic and change its redox state. In this study we show that semiquinone radicals produced during microbial or chemical reduction of a HS model quinone (AQDS, 9,10-anthraquinone-2,6-disulfonic acid) are strong oxidants. They oxidize arsenite to arsenate, thus decreasing As toxicity and mobility. This reaction depends strongly on pH with more arsenite (up to 67.3%) being oxidized at pH 11 compared to pH 7 (12.6% oxidation) and pH 3 (0.5% oxidation). In addition to As(III) oxidation by semiquinone radicals, hydroquinones that were also produced during quinone reduction reduced As(V) to As(III) at neutral and acidic pH values (less than 12%) but not at alkaline pH. In order to understand redox reactions between arsenite/arsenate and reduced/oxidized HS, we quantified the radical content in reduced quinone solutions and constructed Eh-pH diagrams that explain the observed redox reactions. The results from this study can be used to better predict the fate of arsenic in the environment and potentially explain the occurrence of oxidized As(V) in anoxic environments. PMID- 19544867 TI - Mercury trends in ringed seals (Phoca hispida) from the western Canadian Arctic since 1973: associations with length of ice-free season. AB - We examined a unique time series of ringed seal (Phoca hispida) samples collected from a single location in the western Canadian Arctic between 1973 and 2007 to test for changes in total mercury (THg) in muscle tissue associated with (1) year and (2) length of ice-free season. We found no temporal trend with muscle THg whereas a curvilinear relationship existed with the length of ice-free season: seals attaimed higher THg in short (2 months) and long (5 months) ice-free seasons. delta 15N and delta13C in muscle tissue did not illustrate significant trends with ice-free days. We estimated that the turnover time of THg in muscle was about twice as long as stable isotope turnover in muscle, possibly explaining the lack of trend with stable isotopes in association with ice-free duration. Our discussion explains how summer environmental conditions may influence the composition of prey (mercury exposure) available to ringed seals. Results offer insight into how marine mammals may respond to directional changes in the Arctic ice-free season. PMID- 19544868 TI - Microbial chlorination of organic matter in forest soil: investigation using 36Cl chloride and its methodology. AB - Chloride, which comes into the forest ecosystem largely from the sea as aerosol (and has been in the past assumed to be inert), causes chlorination of soil organic matter. Studies of the chlorination showed that the content of organically bound chlorine in temperate forest soils is higher than that of chloride, and various chlorinated compounds are produced. Our study of chlorination of organic matter in the fermentation horizon of forest soil using radioisotope 36Cl and tracer techniques shows that microbial chlorination clearly prevails over abiotic, chlorination of soil organic matter being enzymatically mediated and proportional to chloride content and time. Long-term (>100 days) chlorination leads to more stable chlorinated substances contained in the organic layer of forest soil (overtime; chlorine is bound progressively more firmly in humic acids) and volatile organochlorines are formed. Penetration of chloride into microorganisms can be documented by the freezing/thawing technique. Chloride absorption in microorganisms in soil and in litter residues in the fermentation horizon complicates the analysis of 36Cl-chlorinated soil. The results show that the analytical procedure used should be tested for every soil type under study. PMID- 19544869 TI - Electrochemical evidences for promoted interfacial reactions: the role of Fe(II) adsorbed onto gamma-Al2O3 and TiO2 in reductive transformation of 2-nitrophenol. AB - This study was aimed at elucidating the role of adsorbed Fe(II) on minerals in the reductive transformation of 2-nitrophenol (2-NP) by using electrochemical methods. The studies of Fe(ll) adsorption and 2-NP reduction kinetics showed that the identity of minerals such as gamma-Al2O3 and TiO and the solution pH were crucial factors to determine the Fe(ll) adsorption behavior and to influence the rate constant (k) of 2-NP reduction. Furthermore, two electrochemical methods, cyclic voltammetry (CV) and electrochemical impedance spectrometry (EIS), were applied to characterize the Fe(II) reactivity with both the mineral-coated and mineral-free electrodes. The electrochemical evidence confirmed that the peak oxidation potential (Ep) of complex Fe(II) can be significantly affected by the solution pH;the enhanced reductive transformation of 2-NP can be related to the reduced Ep of surface-complex Fe(II) and the reduced charge transfer resistance (R(CT)) of the Fe(III)/Fe(II) couple. All these relationships were studied quantitatively. At pH 6.7, the measured Ep and R(CT) decreased in the order TiO2/GC < gamma-Al2O3/ GC < GC (Ep, 0.140 < 0.190 < 0.242 V; R(CT), 0.30 < 0.41 < 0.78 komega), while the 2-NP reduction on different minerals were in the order TiO2 > gamma-Al2O3 > nonmineral (k x 10-2, 7.91 > 0.64 > 0.077 min(-l)). PMID- 19544870 TI - Indirect photolysis of perfluorochemicals: hydroxyl radical-initiated oxidation of N-ethyl perfluorooctane sulfonamido acetate (N-EtFOSAA) and other perfluoroalkanesulfonamides. AB - Selected perfluorinated surfactants were irradiated in aqueous hydrogen peroxide solutions using artificial sunlight to study transformation under aquatic environmental conditions. Indirect photolysis mediated by hydroxyl radical was observed for N-ethyl perfluorooctane sulfonamidoethanol (N-EtFOSE), N-ethyl perfluorooctane sulfonamido acetate (N-EtFOSAA), N-ethyl perfluorooctane sulfonamide (N-EtFOSA), and perfluorooctane sulfonamide acetate (FOSAA). An upper limitforthe bimolecular reaction rate constant for reaction of *OH and N-EtFOSAA was determined to be (1.7 +/- 0.7) x 10(9) M(-1)s(-1). A proposed reaction pathwayfor degradation of the parent perfluorochemical, N-EtFOSE, to the other perfluoroalkanesulfonamides and perfluorooctanoate (PFOA) was developed and includes oxidation and N-dealkylation steps. As they did not undergo additional degradation, perfluorooctane sulfonamide (FOSA) and PFOA were the final degradation products of hydroxyl radical-initiated oxidation. UV-visible absorption spectra for the perfluorochemicals, showing absorbance in the UV region below the range of natural sunlight are also reported. In sunlit environments, indirect photolysis of perfluorochemicals is likely to be important in the determination of their environmental fate given the slow rates expected for biotransformation and weak sorption. Photolytic conversion of perfluorochemicals into refractory perfluorinated acids, mainly PFOA, could mean that a significant fraction of these compounds will accumulate in the world's oceans. PMID- 19544872 TI - Influence of magnetite stoichiometry on Fe(II) uptake and nitrobenzene reduction. AB - Magnetite (Fe3O4) is a common biomineralization product of microbial iron respiration and is often found in subsurface anoxic environments, such as groundwater aquifers where aqueous Fe(II) is present We investigated the reaction between aqueous Fe(II) and magnetite using the isotopic selectivity of 57Fe Mossbauer spectroscopy and revisited the reduction of nitrobenzene by magnetite. Similar to our previous findings with Fe3+ oxides, we did not observe the formation of a stable sorbed Fe(II) species; instead, we observed oxidation of the Fe(II) to a partially oxidized magnetite phase. Oxidation of Fe(II) was accompanied by reduction of the octahedral Fe3+ atoms in the underlying magnetite to octahedral Fe2+ atoms. The lack of a stable, sorbed Fe(II) species on magnetite prompted us to reevaluate what is controlling the extent of Fe(II) uptake on magnetite, as well as contaminant reduction in the presence of magnetite and Fe(II). Uptake of Fe(II) by magnetite appears to be limited by the stoichiometry of the magnetite particles, rather than the surface area of the particles. More oxidized (or less stoichiometric) magnetite particles take up more Fe(II), with the formation of stoichiometric magnetite (Fe2+/Fe3+ = 0.5) limiting the extent of Fe(II) uptake. We also showthat stoichiometric magnetite, in the absence of aqueous Fe(II), can rapidly reduce nitrobenzene. Based on these results, we speculate that contaminant reduction that was previously attributed to Fe(II) sorbed on magnetite is due to a process similar to negative (n) doping of a solid, which increases the stoichiometry of the magnetite and alters the bulk redox properties of the particle to make reduction more favorable. PMID- 19544871 TI - Effect of bioirrigation on sediment-water exchange of methylmercury in Boston Harbor, Massachusetts. AB - Coastal marine sediments are important sites of methylmercury (MMHg) production, and dissolved efflux provides an important source of MMHg to near-shore, and possibly offshore, water columns and food webs. We measured the flux of MMHg across the sediment-water interface at four stations in Boston Harbor that span a range of infaunal population densities and bioirrigation intensities. At each station we carried out total MMHg flux measurements using core incubations and collected near-surface pore waters to establish MMHg gradients for diffusive flux calculations. The flux cores were also imaged by CT scanning to determine the distribution of infaunal burrows, and pore-water sulfide and 222Rn profiles were measured. Total MMHg fluxes, measured using core incubations, ranged from -4 to 191 pmol m(-2) d(-1), and total MMHg fluxes were strongly correlated with burrow densities at the stations. Estimated diffusive fluxes, calculated based on MMHg concentration gradients below the sediment-water interface, were much lower than total fluxes at three of the stations, ranging from 2-19 pmol m(-2) d(-1). These results indicate that MMHg exchange may be significantly enhanced over molecular diffusion in bioturbated sediments. Furthermore, burrow density provides a strong predictor of total MMHg flux. Pore-water exchange of both dissolved MMHg and 222Rn, a naturally occurring pore-watertracer, increased across the range of observed burrow densities, suggesting that the presence of burrows enhances both MMHg production and flux. PMID- 19544873 TI - Effect of suspended-sediment concentration on nitrification in river water: importance of suspended sediment-water interface. AB - High suspended sediment (SPS) concentrations exist in many Asian river systems. In this research, the effects of SPS concentration on nitrification in river water systems were studied. With orwithout introducing ammonium-oxidizing bacteria isolated from the water and sediment samples of the Yellow River, the microbially mediated nitrification rate increased with SPS concentration as described by the power function y = a x x(b), where y is the nitrification rate, x is the SPS concentration, and a and b are constants. With an indigenous ammonium-oxidizing bacteria, nitrification rate constants, i.e., K4 (micromax/Ks) values obtained from the Monod model, were 0.0016, 0.0036, 0.0040, 0.0063, 0.0066, 0.0071, and 0.0077 day(-1) microM(-1) for the systems with SPS concentrations of 0, 0.2 1.0, 5.0, 10, 20, and 40 g/L, respectively. The sorption percentage of NH4+-N increased with SPS concentration as a power function. Bacteria tend to attach onto SPS, and the maximum specific growth rate at the SPS water interface was about twice that in the water phase. An increase of bacterial population and nitrification rate constant with SPS as a power function resulted in an increase of nitrification rate with SPS as a power function. Therefore, the high SPS concentration caused by erosion and bottom sediment resuspension and other factors will accelerate ammonium oxidation in many turbid river systems. This has useful implications for nitrogen removal from river systems. PMID- 19544874 TI - Accumulation, subcellular distribution and toxicity of copper in earthworm (Eisenia fetida) in the presence of ciprofloxacin. AB - Land application of wastes from concentrated animal feeding operations results in accumulation of copper (Cu) and antimicrobials in terrestrial systems. Interaction between Cu and antimicrobials may change Cu speciation in soil solution, and affect Cu bioavailability and toxicity. In this study, earthworms were exposed to quartz sand percolated with different concentrations of Cu and ciprofloxacin (CIP). Copper uptake by earthworms, its subcellular partition, and toxicity were studied. An increase in the applied CIP decreased the free Cu ion concentration in external solution and mortalities of earthworm, while Cu contents in earthworms increased. Copper and CIP in earthworms were fractionated into five fractions: a granular fraction (D), a fraction consisting of tissue fragments, cell membranes, and intact cells (E), a microsomal fraction (F), a denatured proteins fraction (G), and a heat-stable proteins fraction (H). Most of the CIP in earthworms was in fraction H. Copper was redistributed from the metal sensitive fraction E to fractions D, F, G, and H with increasing CIP concentration. These results challenge the free ion activity model and suggested that Cu may be partly taken up as Cu-CIP complexes in earthworms, changing the bioavailability, subcellular distribution, and toxicity of Cu to earthworms. PMID- 19544875 TI - An agglomeration-based model for colloid filtration. AB - This paper develops a model for colloid filtration which accounts for the possibility of aggregation of the colloidal particles in the aqueous phase. Depth wise variation of liquid-phase colloid concentration, C(x), is measured experimentally in the presence of monovalent cations at different concentrations and divalent cations, which confirm that log of C(x) deviates significantly from linearity for all of the above cases. It is also observed that in all systems showing significant removal, preaggregation of the colloids is observed, which confirms the correlation between aggregation of colloids with their retention in saturated porous media, which has been reported earlier. A new model for depth filtration which is based on material balances of the different sized aggregates, which could be present in the colloidal dispersion, is developed. This resulting model, based on differential deposition rates for different sized aggregates, shows good match with the experimentally observed variation of C(x) with depth, for all of the above conditions. A method for estimating model parameters from data is developed. This model remains independent of the actual deposition mechanism, which could be secondary minima attachment of these colloidal aggregates or the entrapment of these colloidal aggregates in regions of confined geometry. This model can be extended to predict the transport of colloids in groundwater. PMID- 19544876 TI - Patterns of bioaccumulation of polybrominated diphenyl ether and polychlorinated biphenyl congeners in marine mussels. AB - Marine mussels (Modiolus modiolus) and sediment from 14 stations near a municipal outfall and three reference locations were analyzed for polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) to evaluate and compare patterns of bioaccumulation of individual congeners between these two groups of chemicals. Of the 47 PBDEs and 209 PCBs analyzed, 34 PBDE and 153 PCB congeners or coeluting groups of congeners were detected in one or more matrices. The predominant PBDE congeners were BDEs 47, 99, 100, and 209, accounting for 80-90% of the total PBDEs in all matrices. PCBs and PBDEs exhibited a parabolic relationship of the biota-sediment accumulation factor (BSAF) versus the log octanol-water partition coefficient(Kow). Below Kow 10(5.5), BSAFs ranged between 1 and 3, reflecting approximate equilibrium between mussels and sedimentforthese relatively water soluble congeners. BSAFs increased with increasing Kow to maximum values of approximately 30-100 for congeners with Kow approximately 10(7) and then declined at higher Kow to a value of approximately 1 for BDE 209. BSAFs for PBDEs were generally 2- to 3-fold higher than those for PCBs of a similar Kow. The calculated BSAFs for PBDE congeners indicate that PBDEs have a pattern of bioaccumulative behavior in mussels similar to that of the PCBs, and that some PBDE congeners may be more bioaccumulative in mussels than PCBs. PMID- 19544877 TI - Integrated stochastic environmental risk assessment of the harbour area treatment scheme (HATS) in Hong Kong. AB - Submarine ocean outfalls are commonly used for the disposal of partially treated effluents in coastal cities. Typically, the greatest environmental risk caused by toxic substances occurs in the near field of the outfall discharge. The ecological impact of the effluent varies greatly under different discharge and environmental conditions that are characterized by both regular and stochastic variations. For a comprehensive environmental risk assessment of a coastal discharge, it is necessary to determine both the likelihood and severity of the adverse effects on the biological community. We present the first integrated stochastic (Monte Carlo) environmental risk assessment of a major coastal sewage outfall discharge--the Stonecutters Island outfall of the Harbour Area Treatment Scheme (HATS) in Hong Kong. Unionized ammonia (NH3) is used as the target pollutant. To accurately envisage the ambient concentrations of NH3, a Lagrangian jet model (JETLAG/VISJET) is used to analyze pollutant concentrations in the nearfield of the outfall. The environmental conditions are simulated from 3D hydrodynamic model simulations over a 4 month period for typical wet and dry seasons. Statistical characteristics of the effluent discharge and receiving water temperature are derived from field data. The probability distribution of predicted exposure concentrations (EC) is generated from this integrated simulation. A species sensitivity distribution, which represents a statistical distribution of threshold sublethal effects levels or benchmark concentrations (BC) for various marine organisms is constructed using available chronic toxicity data. The environmental risk of NH3 on the marine community is characterized by computing statistical distributions of Hazard Quotient (HO = EC/BC) using Monte Carlo simulation. It is found that the probability of HO > for HATS Stage 1 (1.6 million m3/day sewage treated with chemically enhanced primary treatment) is around 0.11 for wet season but just about 0.06 for the dry season. The risk increases by about 10% to 0.08-0.13 with additional sewage loads of 0.8 million m3/day at the same level of treatment (HATS Stage 2A). With an upgrade to secondary treatment (HATS Stage 2B), the probability will be reduced to 0.03 0.05. Compared to the use of "worst case" scenarios or point pollution threshold estimates, the present method offers a more holistic ecological assessment, and is much less sensitive to arbitrary choice of model parameters. The present risk assessment approach can be readily extended to the accurate determination of mixing zones based on statistical evaluation of ecological risks. PMID- 19544878 TI - Surface association of motile bacteria at granular porous media interfaces. AB - Bacterial populations were observed using dark-field light scattering at porous media interfaces comprised of a dilute solution containing the polymer additives methylcellulose and a transparent particulate suspension composed of mechanically agitated Gelrite gellan gum. Population-scale experiments with a nonchemotactic smooth-swimming mutant, Escherichia col HCB 437, yielded a variety of distinct and reproducible bacterial distributions that included highly concentrated bands of bacteria near the interface. While no physical attachment was observed between the bacteria and granular Gelrite media, the population exhibited surface associations characterized by reversible physical obstructions of the motile bacteria at the solid granular surfaces. These interactions decreased translational motion, which reduced bacterial migration and concentrated bacterial populations near the interface. Results from glass bead experiments indicated similar surface associations in high-surface area glass bead environments. Experimental results were semiquantitatively analyzed using a one dimensional population-scale transport model. Theoretical profiles were generated using a single set of parameters and simultaneously compared with averaged bacterial distributions from multiple interface configurations. Parameter estimates were consistent with expected values. The agreement between the theoretical and experimental data suggests a quantifiable approach for modeling bacterial migration within high-surface area granular media environments. PMID- 19544879 TI - Cost-effective hydraulic tomography surveys for predicting flow and transport in heterogeneous aquifers. AB - This study shows how a cost-effective hydraulic tomography survey (HTS) and the associated data estimator can be used to characterize flow and transport in heterogeneous aquifers. The HTS is an improved field hydraulic test that accounts for responses of hydraulic stresses caused by pumping or injection events at different locations of an aquifer. A sequential data assimilation method based on a cokriging algorithm is then used to map the aquifer hydraulic conductivity (K). This study uses a synthetic two-dimensional aquifer to assess the accuracy of predicted concentration breakthrough curves (BTCs) on the basis of the Kfields estimated by geometric mean, kriging, and HTS. Such Kfields represent different degrees of flow resolutions as compared with the synthetically generated one. Without intensive experimentsto calibrate accurate dispersivities at sites, the flow field based on the HTS Kfield can yield accurate predictions of BTC peaks and phases. On the basis of calculating mean absolute and square errors for estimated K fields, numerical assessments on the HTS operation strategy show that more pumping events will generally lead to more accurate estimations of Kfields, and the pump locations need to be installed in high Kzones to maximize the delivery of head information from pumps to measurement points. Additionally, the appropriate distances of installed wells are suggested to be less than one-third of the ln(K) correlation length in x direction. PMID- 19544880 TI - Space/time analysis of fecal pollution and rainfall in an eastern North Carolina estuary. AB - The Newport River Estuary (NPRE) is a high-priority shellfish harvesting area in eastern North Carolina that is impaired due to fecal contamination, specifically exceeding recommended levels for fecal coliforms. A hydrologic-driven mean trend model was developed, as a function of antecedent rainfall, in the NPRE to predict levels of Escherichia coli (EC, measured as a proxyforfecal coliforms). This mean trend model was integrated in a Bayesian Maximum Entropy (BME) framework to produce informative space/time (S/T) maps depicting fecal contamination across the NPRE during winter and summer months. These maps showed that during dry winter months, corretponding to the oyster harvesting season in North Carolina (October 1-March 30), predicted EC concentrations were below the shellfish harvesting standard (14 MPN/100 mL). However, after substantial rainfall of 3.81 cm (1.5 in.), the NPRE did not appear to mee this requirement. Warmer months resulted in the predicted EC concentrations exceeding the threshold for the NPRE. Predicted ENT concentrations were generally below the recreational water quality threshold (104 MPN/100 mL), except for warmer months after substantial rainfall. Once established, this combined approach produces near real-time visual information on which to base water quality management decisions. PMID- 19544881 TI - Modern space/time geostatistics using river distances: data integration of turbidity and E. coli measurements to assess fecal contamination along the Raritan River in New Jersey. AB - Escherichia coli (E. coli) is a widely used indicator of fecal contamination in water bodies. External contact and subsequent ingestion of bacteria coming from fecal contamination can lead to harmful health effects. Since E. coli data are sometimes limited, the objective of this study is to use secondary information in the form of turbidity to improve the assessment of E. coli at unmonitored locations. We obtained all E. coli and turbidity monitoring data available from existing monitoring networks for the 2000-2006 time period for the Raritan River Basin, New Jersey. Using collocated measurements, we developed a predictive model of E. coli from turbidity data. Using this model, soft data are constructed for E. coli given turbidity measurements at 739 space/time locations where only turbidity was measured. Finally, the Bayesian Maximum Entropy (BME) method of modern space/time geostatistics was used for the data integration of monitored and predicted E. coli data to produce maps showing E. coli concentration estimated daily across the river basin. The addition of soft data in conjunction with the use of river distances reduced estimation error by about 30%. Furthermore, based on these maps, up to 35% of river miles in the Raritan Basin had a probability of E coli impairment greater than 90% on the most polluted day of the study period. PMID- 19544882 TI - A novel approach to evaluate the production kinetics of extracellular polymeric substances (EPS) by activated sludge using weighted nonlinear least-squares analysis. AB - This paper develops a novel and convenient approach for evaluation of production kinetics of extracellular polymeric substances (EPS) by activated sludge. In this approach, the weighted least-squares analysis is employed to calculate approximate differences in EPS concentration between model predictions and data. An iterative search routine in the Monte Carlo method is utilized for optimization of the objective function by minimizing the sum of squared weighted errors. Application of the approach in this work shows that the fraction of substrate electrons diverted to EPS formation (k(EPS)) is 0.23 g COD(EPS) g(-1) CODs with a bacterial maximum growth rate of 0.32 h(-1). The obtained parameters are found to be reasonable as they are generally bounded. The validity of this approach is confirmed by both the independent EPS production tests and the EPS data reported in literature. It also corrects the overestimation of cellular production and identifies that k(EPS) is the key parameter in EPS production kinetics. Furthermore, this approach could estimate the kinetic parameters accurately using few data sets or even one set, which becomes very attractive for the processes where data are costly to obtain. PMID- 19544883 TI - Addressing temporal variability when modeling bioaccumulation in plants. AB - Steady state models are commonly used to predict bioaccumulation of organic contaminants in biota. However, the steady state assumption may introduce errors when complex dynamic processes such as growth, temperature fluctuations, and variable environmental concentrations significantly affect the major chemical uptake and elimination processes. In this study, a strategy for addressing temporal variability in bioaccumulation modeling is proposed. Chemical partitioning space plots are used to show the time necessary for organic contaminants to approach steady state in plant leaves and roots as well as the dominant uptake/elimination fluxes of chemicals as a function of the contaminants' physical chemical properties. The plots were produced with a novel nonsteady state model of bioaccumulation in plants, which is presented, parameterized, and evaluated. The first prerequisite identified for using a steady state model is that the duration of chemical exposure exceeds the time to approach steady state. Next, the dominant chemical transport processes for the chemical in question should be identified and the variability of parameters affecting these processes compared to the time to approach steady state. A major systematic variation in one of these parameters on a time scale similar to the time to approach steady state may cause an unacceptable deviation between the predicted and true chemical concentrations in vegetation. In such cases a nonsteady state model such as the one presented here should be used. The chemical partitioning plots presented provide guidance for understanding the dominant uptake/elimination processes and the time to approach steady state in relation to the partitioning properties of organic compounds. PMID- 19544884 TI - PAH bioavailability in field sediments: comparing different methods for predicting in situ bioaccumulation. AB - In situ exposure concentrations of chemicals in sediments and their depending risks are determined by site-specific parameters (e.g., sediment organic carbon composition), controlling bioavailability. Over the years, several analytical methods have been developed to assess bioavailable concentrations or fractions. Some of these methods have been successful in the laboratory, but few attempts have been made to test their potential for predicting actual in situ bioavailability. In this study, solid-phase microextraction (SPME)-fibers and aquatic worms (Lumbriculus variegatus) were exposed in situ at three locations. In addition, laboratory-based methods, i.e., methods with which sampling of the bioavailable fraction/concentration took place in the laboratory, being SPME, polyoxymethylene solid-phase extraction (POM-SPE), hydroxypropyl-beta-yclodextrin (HPCD), and 6 h-Tenax extraction were applied to sediments collected at the locations. Using equilibrium partitioning-based calculations, biotic PAH levels were calculated from the concentrations or fractions extracted by the used methods. In general, method-predicted concentrations were within a factor of 10 of those measured in field-exposed oligochaetes, with in situ SPME and laboratory based POM-SPE yielding the best results. As a reference, the currently used generic risk assessment approach overpredicted biotic concentrations by a factor of 10-100, which corresponded to in situ SPME-derived sediment-water distribution coefficients and biota-to-sediment accumulation factors being up to 2 orders of magnitude higher and lower, respectively, than generic values. These observations advocate site-specific risk assessment for PAHs, for which potential tools were evaluated in this study. PMID- 19544885 TI - Molecular characterization of nitrogen-containing organic compounds in biomass burning aerosols using high-resolution mass spectrometry. AB - Although nitrogen-containing organic compounds (NOC) are important components of atmospheric aerosols, little is known about their chemical composition. Here we present detailed characterization of the NOC constituents of biomass burning aerosol (BBA) samples using high-resolution electrospray ionization mass spectrometry (ESI/MS). Accurate mass measurements combined with MS/MS fragmentation experiments of selected ions were used to assign molecular structures to individual NOC species. Our results indicate that N-heterocyclic alkaloid compounds (species naturally produced by plants and living organisms) comprise a substantial fraction of NOC in BBA samples collected from test burns of five biomass fuels. High abundance of alkaloids in test burns of ponderosa pine (a widespread tree in the western U.S. areas frequently affected by large scale fires) suggests that N-heterocyclic alkaloids in BBA may play a significant role in dry and wet deposition of fixed nitrogen in this region. PMID- 19544886 TI - Simultaneous measurement of free and total concentrations of hydrophobic compounds. AB - Hydrophobic contaminants partition between water and sorbent phases such as dissolved organic matter (DOM) in environmental matrices. Consideration of bioavailability increasingly requires the measurement of freely dissolved concentration (Cfree) in addition to the total chemical concentration (Ctotal). However, current practices require the use of separate methods and samples to derive Cfee and Ctotal, and no method affords simultaneous measurementof both concentrations. We employed a method by coupling solid-phase microextraction (SPME) with the use of an isotope-labeled internal standard and gas chromatography-mass spectrometer (GC-MS) for detection. The method was demonstrated using 8 pyrethroids as model compounds and 13C-cis-permethrin as the labeled internal standard. Analysis of 4 spiked sediment porewater and 4 field contaminated samples showed that Ctotal obtained by this method was in close agreementwith the spiked concentrations or the total concentrations measured after whole-sample solvent extraction, while Cfree was only a small fraction of Ctotal, suggesting pronounced association of these compounds with DOM. The method displayed good reproducibility with average relative standard deviation <20%, and high sensitivity with method detection limits in the range of 1-5 ng L(-1). The method uses a small sample volume (<20 mL, as opposed to 1 L for conventional analysis), and is solventless and fast. The concurrent measurements also allow for estimation of K(DOC), a key parameter that is difficult to derive otherwise. The simultaneous determination of Cfree and Ctotal provides information for better relating chemical concentrations with potential ecotoxicological effects. PMID- 19544887 TI - Mitigation of arsenic accumulation in rice with water management and silicon fertilization. AB - Rice represents a major route of As exposure in populations that depend on a rice diet. Practical measures are needed to mitigate the problem of excessive As accumulation in paddy rice. Two potential mitigation methods, management of the water regime and Si fertilization, were investigated under greenhouse conditions. Growing rice aerobically during the entire rice growth duration resulted in the leastAs accumulation. Maintaining aerobic conditions during either vegetative or reproductive stage of rice growth also decreased As accumulation in rice straw and grain significantly compared with rice grown under flooded conditions. The effect of water management regimes was consistent with the observed effect of flooding-induced arsenite mobilization in the soil solution. Aerobic treatments increased the percentage of inorganic As in grain, but the concentrations of inorganic As remained lower than in the flooded rice. Silicon fertilization decreased the total As concentration in straw and grain by 78 and 16%, respectively, even though Si addition increased As concentration in the soil solution. Silicon also significantly influenced As speciation in rice grain and husk by enhancing methylation. Silicon decreased the inorganic As concentration in grain by 59% while increasing the concentration of dimethylarsinic acid (DMA) by 33%. There were also significant differences between two rice genotypes in grain As speciation. This study demonstrated that water management Si fertilization, and selection of rice cultivars are effective measures that can be used to reduce As accumulation in rice. PMID- 19544888 TI - Iodide-mediated photooxidation of arsenite under 254 nm irradiation. AB - The preoxidation of As(III) to As(V) is a desirable process to increase the removal efficiency of arsenic in water treatment In this work, the photooxidation of As(III) under 254 nm irradiation was investigated in the concentration range of 1-1000 microM in the presence of potassium iodide (typically 100 microM). Although the direct photooxidation of As(III) in water was negligible, the presence of iodide dramatically enhanced the oxidation rate. The quantitative conversion of As(III) to As(V) was achieved. The quantum yields of As(III) photooxidation ranged from 0.08 to 0.6, depending on the concentration of iodide and As(III). The excitation of iodides under 254 nm irradiation led to the generation of iodine atoms and triiodides, which seem to be involved in the oxidation process of As(III). Because the efficiency of iodine atom generation is highly dependent on the presence of suitable electron acceptors,the photooxidation of As(III) was efficient in an air- or N2O-saturated solution but markedly reduced in the N2-saturated solution. The production of H2O2 was also accompanied by the generation of As(V). The addition of excess methanol (OH radical scavenger) did not reduce the photooxidation rate at all, which ruled out the possibility of hydroxyl radical involvement. It was found that the in situ photogenerated triiodides oxidize As(III) with regenerating iodides by completing a cycle. The proposed UV254/KI/As(III) process is essentially an iodide-mediated photocatalysis. PMID- 19544889 TI - Diagnosis of treatment efficiency in industrial wastewater treatment plants: a case study at a refinery ETP. AB - Many industries employ the activated sludge process for biological removal of pollutants present in wastewater. Yet, treatment plants do notfunction at optimum potential. The biological component of such systems remains a black box, and reasons responsible for poor performance have not been identified. We have used genomic and physiological tools to understand the process and propose that analysis of catabolic signatures and nutrient levels, are crucial parameters in assessing and monitoring the performance of an effluent treatment plant. In this study, we use activated sludge collected from a refinery running at a capacity of 8 million metric tonnes of wastewater as a model. The presence of hydroxylases, oxygenases, and dioxygenases in the biomass was demonstrated by polymerase chain reaction and sequence analysis of aromatic-ring hydroxylating dioxygenase clones extracted from the metagenome, suggests the presence of hitherto unreported enzymes. The actual degradative state of the biomass was demonstrated by respirometric analysis using 11 substrates expected in refinery wastewater. Nutrient-levels required for the microbial population were estimated by on-site analysis. Diagnosis of the degradative potential of activated sludge can be carried out by incorporating these tools in regular monitoring procedures and can setthe rules for improving the efficiency of treatment PMID- 19544890 TI - Improvement of electrochemical wastewater treatment through mass transfer in a seepage carbon nanotube electrode reactor. AB - A seepage carbon nanotube electrode (SCNE) reactor was designed in order to facilitate contaminant mass transfer from bulk solution to the electrode surface, therefore to break the high cost bottleneck of electrochemical wastewater treatment. The innovative concept behind the reactor design is that the overall mass transfer would be significantly improved via contaminant migration through the porous carbon nanotube electrode. It was found out that the surface diffusivity D(s,i) in the external film was the controlling coefficient for electrochemical treatment, and the proposed process could improve the overall mass transfer coefficient by 116-161% compared with conventional electrochemical reactors under the same conditions. The research also showed that the current efficiency of the SCNE reactor was 340-519% higher than that of conventional reactors, and the energy consumption to mineralize the same amount of organics was only 16.5-22.3% of the conventional reactors. Also, the influences of potential, pH, and electrolyte concentration were investigated to optimize the operating parameters for the SCNE reactor. These results show that the SCNT reactor is promising because of its energy efficiency and has potential for application in wastewater treatment PMID- 19544891 TI - Effect of adsorbed polyelectrolytes on nanoscale zero valent iron particle attachment to soil surface models. AB - Polyelectrolyte coatings significantly increase the mobility of nanoscale zerovalent iron (NZVI) in saturated porous media. The effect can be attributed to improved colloidal stability of NZVI suspensions, decreased adhesion to soil surfaces, or a combination of the two effects. This research explicitly examines how coatings control NZVI adhesion to model soil surfaces. NZVI was coated with three different polyeleotrolyte block copolymers based on poly(methacrylic acid), poly(methyl methacrylate or butyl methacrylate), and poly(styrenesulfonate) or with a poly(styrenesulfonate) homopolymer. SiO2 and a humic acid film served as model soil surfaces. The polyelectrolytes increased the magnitude of the electrophoretic mobility of NZVI over a broad pH range relative to unmodified NZVI and shifted the isoelectric point outside the typical groundwater pH range. Quartz crystal microgravimetry measurements indicated extensive adhesion of unmodified NZVI to SiO2. Polyelectrolyte coatings decreased adhesion by approximately 3 orders of magnitude. Adding 50 mM NaCL to screen electrostatic repulsions did not significantly increase adhesion of modified NZVI. Coated NZVI did not adhere to humic acid films for either 1 mM NaHCO3 or 1 mM NaHCO3 + 50 mM NaCl. The lack of adhesion even in a high ionic strength medium was attributed to electrosteric repulsion, as opposed to electrostatic double layer repulsion, between the polyelectrolyte-coated NZVI and the negatively charged surfaces. The lack of significant adhesion on either model surface was observed for all polymer architectures investigated. PMID- 19544892 TI - Active filtration of phosphorus on Ca-rich hydrated oil shale ash: does longer retention time improve the process? AB - The purpose of this research was to investigate the phosphorus binding capacity of active filtration through hydrated Ca-rich oil shale ash sediment in a laboratory-scale experiment at different retention times. Three column experiments were established with hydraulic loading rates of 12 L d(-1), 18 L d( 1), and 24 L d(-1), with residence times of 12, 8, and 6 h, respectively. The results showed good removal efficiency (up to 91% at loading of 1.66 gP m(-2) d( 1)) in the experiment with the longest retention time. The overall performance of the filters did not, however, improve with increasing retention time, and the decrease in binding capacity was 41% at longer retention times. Chemical clogging by carbonate precipitates probably reduces the availability of Ca from the dissolution of unstable ash sediment mineral phases, and the phosphate removal decreases rapidly from 91% to 49% at loading of 1.66 gP m(-2) d(-1). PMID- 19544893 TI - Field application of activated carbon amendment for in-situ stabilization of polychlorinated biphenyls in marine sediment. AB - We report results on the first field-scale application of activated carbon (AC) amendment to contaminated sediment for in-situ stabilization of polychlorinated biphenyls (PCBs). The test was performed on a tidal mud flat at South Basin, adjacent to the former Hunters Point Naval Shipyard, San Francisco Bay, CA. The major goals of the field study were to (1) assess scale up of the AC mixing technology using two available, large-scale devices, (2) validate the effectiveness of the AC amendment at the field scale, and (3) identify possible adverse effects of the remediation technology. Also, the test allowed comparison among monitoring tools, evaluation of longer-term effectiveness of AC amendment, and identification of field-related factors that confound the performance of in situ biological assessments. Following background pretreatment measurements, we successfully incorporated AC into sediment to a nominal 30 cm depth during a single mixing event, as confirmed by total organic carbon and black carbon contents in the designated test plots. The measured AC dose averaged 2.0-3.2 wt% and varied depending on sampling locations and mixing equipment. AC amendment did not impact sediment resuspension or PCB release into the water column over the treatment plots, nor adversely impactthe existing macro benthic community composition, richness, or diversity. The PCB bioaccumulation in marine clams was reduced when exposed to sediment treated with 2% AC in comparison to the control plot Field-deployed semi permeable membrane devices and polyethylene devices showed about 50% reduction in PCB uptake in AC-treated sediment and similar reduction in estimated pore-water PCB concentration. This reduction was evident even after 13-month post-treatment with then 7 months of continuous exposure, indicating AC treatment efficacy was retained for an extended period. Aqueous equilibrium PCB concentrations and PCB desorption showed an AC-dose response. Field-exposed AC after 18 months retained a strong stabilization capability to reduce aqueous equilibrium PCB concentrations by about 90%, which also supports the long-term effectiveness of AC in the field. Additional mixing during or after AC deployment, increasing AC dose, reducing AC-particle size, and sequential deployment of AC dose will likely improve AC-sediment contact and overall effectiveness. The reductions in PCB availability observed with slow mass transfer under field conditions calls for predictive models to assess the long term trends in pore-water PCB concentrations and the benefits of alternative in situ AC application and mixing strategies. PMID- 19544894 TI - Fe0 nanoparticles remain mobile in porous media after aging due to slow desorption of polymeric surface modifiers. AB - Nanosized zerovalent iron (nZVI) is used for in situ remediation of contaminated groundwater. Polyelectrolyte surface coatings are used to inhibit nZVI aggregation and enhance mobility in the subsurface for emplacement. The fate of nZVI is of interest given the uncertainties regarding the effects of nanomaterials on the environment, and depends in part on the stability of these surface coatings against desorption and biodegradation. This study measured the rate and extent of desorption of polyelectrolyte coatings used to stabilize nZVI, including polyaspartate (PAP MW = 2.5 kg/mol and 10 kg/mol), carboxymethyl cellulose (CMC MW= 90 kg/nol and 700 kg/ mol), and polystyrene sulfonate (PSS MW = 70 kg/mol and 1000 kg/mol). The initial adsorbed mass of polyelectrolyte ranged from 0.85 to 3.71 mg/m2 depending on the type and molecular weight (MW). Polyelectrolyte adsorption was confirmed by an increase in nZVI electrophoretic mobility. In all cases, desorption of polyelectrolyte was slow, with less than 30 wt% desorbed after 4 months. The higher MW polyelectrolyte had a greater adsorbed mass and a slower desorption rate for PAP and CMC. nZVI mobility in sand columns after 8 month of desorption was similar to freshly modified nZVI, and significantly greater than unmodified nZVI aged for the same time under identical conditions. Based on these results, polyelectrolyte modified nanoparticles will remain more mobile than their unmodified counterparts even after-aging. Other factors potentially affecting the fate of coated nZVI must be evaluated, especially the potential for biodegradation of coatings. PMID- 19544895 TI - Ferrate (Fe(VI)) application for Municipal wastewater treatment: a novel process for simultaneous micropollutant oxidation and phosphate removal. AB - A novel technology for enhanced municipal wasterwater treatment was assessed based on the dual functions of Fe(VI) to oxidize micropollutants and remove phosphate by formation of ferric phosphates. Second-order rate constants (k) for the reactions of selected pharmaceuticals, endocrine disruptors, and organic model compounds with Fe(VI) were in the range of 1 (trimethylamine) to 9000 M(-1) s(-1) (aniline) in the pH-range 7-8. The selected compounds contained electron rich moieties (ERM) such as phenols, anilines, amines, and olefins. Oxidation experiments in wastewater spiked with micropollutants at concentrations in the low microM range at pH 7 and 8 showed that Fe(VI) doses higher than 5 mg Fe L(-1) are capable of eliminating various ERM-containing micropollutants by more than 85%. In comparison to ozone, Fe(VI) was as effective or slightly less effective in terms of micropollutants oxidation, with Fe(VI) having the benefit of phosphate removal. To lower phosphate from 3.5 to 0.8 mg PO4-P L(-1) (regulatory limit for wastewater discharge in Switzerland), a Fe(VI) dose of 7.5 mg Fe L(-1) was needed. Overall, this study demonstrates Fe(VI) as a promising tool for an enhanced wastewater treatment to remove micropollutants as well as to control phosphate in a single treatment step. PMID- 19544896 TI - Role of sulfur during acetate oxidation in biological anodes. AB - The treatment of wastewater containing sulfides in bioelec-trochemical systems (BES) causes deposition of sulfur on the anode as a result of a solely electrochemical process. In this study, we investigate whether microorganisms can use this sulfur, ratherthan the anode or soluble sulfate, as an electron acceptor for the oxidation of acetate. Our results indicate that microorganisms use electrodeposited sulfur as preferable electron acceptor over the anode and sulfate and produce sulfide irrespective of electrochemical conditions. Bioelectrochemical and biological sulfide generation pathways were studied under different electrochemical conditions. The obtained results show that the sulfide generation rate at open circuit condition (anode potential -235 +/- 5 mV versus standard hydrogen electrode, SHE)was higher in comparison to the electrochemical sulfide generation even at a lower potential of -275 mV (vs SHE), confirming that sulfide is produced through biological processes without any current generation. However, during closed circuit operation, the overall Coulombic efficiency (97% +/- 2%) is not affected as the produced sulfide (originating from the reduction of deposited sulfur) is spontaneously reoxidized to sulfur when a favorable potential is maintained. This confirms the mediator role of sulfur during acetate oxidation in BES. A diagrammatic representation of the mechanism is proposed to characterize the interactions between acetate oxidation and sulfur conversions on the anode. PMID- 19544897 TI - Evaluation of butyl rubber as sorbent material for the removal of oil and polycyclic aromatic hydrocarbons from seawater. AB - Ecological disasters resulting from oil spills have created a great need to find more efficient materials for oil spill cleanup. This research highlights the use of a novel macroporous polymeric material based on butyl rubber (BR) as a sorbent in an oil spill cleanup. The sorption capacity of BR for crude oil and petroleum products is 15-23 g g(-1) as compared to the value of 10-16 g g(-1) obtained using a nonwoven polypropylene (PP), a widely used commercial oil sorbent. BR sorbent is reusable after simple squeezing and its continuous sorption capacity for crude oil is 7.6 g g(-1) in each cycle, about 3 times the capacity of the PP sorbent BR sorbents also remove efficiently polycyclic aromatic hydrocarbons (PAHs) such as acenaphthene and pyrene from seawaters. The results suggest that the rubber sorbents are a better alternative to the widely used PP sorbents by improving the efficiency of oil sorption and the reusability of the sorbent. PMID- 19544898 TI - Ferrous and ferric ion generation during iron electrocoagulation. AB - Our research on arsenate removal by iron electrocoagulation (EC) produced highly variable results, which appeared to be due to Fe2+ generation without subsequent oxidation to Fe3+. Because the environmental technology literature is contradictory with regard to the generation of ferric or ferrous ions during EC, the objective of this research was to establish the iron species generated during EC with iron anodes. Experimental results demonstrated that Fe2+, not Fe3+, was produced at the iron anode. Theoretical current efficiency was attained based on Fe2+ production with a clean iron rod, regardless of current, dissolved-oxygen (DO) level, or pH (6.5-8.5). The Fe2+ remaining after generation and mixing decreased with increasing pH and DO concentration due to rapid oxidation to Fe3+. At pH 8.5, Fe2+ was completely oxidized, which resulted in the desired Fe(OH)3(s)/ FeOOH(s), whereas, at pH 6.5 and 7.5, incomplete oxidation was observed, resulting in a mixture of soluble Fe2+ and insoluble Fe(OH)3(s)/FeOOH(s). When compared with Fe2+ chemical coagulation, a transient pH increase during EC led to faster Fe2+ oxidation. In summary, for EC in the pH 6.5 7.5 range and at low DO conditions, there is a likelihood of soluble Fe2+ species passing through a subsequentfiltration process resulting in secondary contamination and inefficient contaminant removals. PMID- 19544899 TI - Oxidative removal of bisphenol A by manganese dioxide: efficacy, products, and pathways. AB - Bisphenol A (BPA) is a ubiquitous environmental contaminant with endocrine disruption potential. In this study, exploiting the outstanding oxidative capacity of manganese dioxide (delta-MnO2), we explored forthe firsttime the efficacy and mechanisms of BPA removal by MnO2. In aqueous solutions, MnO2 demonstrated an extremely efficient capacity to remove BPA. Nearly all BPA (>99%) was eliminated in 6 min in a pH 4.5 solution initially containing 800 microM MnO2 and 4.4 microM BPA. While humic acid showed negligible inhibition on BPA removal, coexisting metal ions such as Mn2+, Ca2+, Mg2+, and Fe3+ displayed suppressive effects and the inhibitive capacityfollowed the order Mn2+ > Ca2+ > Mg2+ approximately Fe3+. A total of 11 products or intermediates were indentified and a detailed reaction scheme was suggested. The products could be ascribed to a suite of reactions of radical coupling, fragmentation, substitution, and elimination, triggered by the BPA radical formed through electron transfers to MnO2. The exceptional efficiency of MnO2 in removing BPA represents a potential use of MnO2 to treat waters containing phenolic compounds and also suggests a potentially important role of oxide-facilitated abiotic transformations in BPA attenuation in natural soil and sediment environments. PMID- 19544900 TI - High-throughput membrane surface modification to control NOM fouling. AB - A novel method for synthesis and screening of fouling-resistant membrane surfaces was developed by combining a high-throughput platform (HTP) approach together with photoinduced graft polymerization (PGP)forfacile modification of commercial poly(aryl sulfone) membranes. This method is an inexpensive, fast, simple, reproducible, and scalable approach to identify fouling-resistant surfaces appropriate for a specific feed. In this research, natural organic matter (NOM) resistant surfaces were synthesized and indentified from a library of 66 monomers. Surfaces were prepared via graft polymerization onto poly(ether sulfone) (PES) membranes and were evaluated using an assay involving NOM adsorption, followed by pressure-driven-filtration. In this work new and previously tested low-fouling surfaces for NOM are identified, and their ability to mitigate NOM and protein (bovine serum albumin)fouling is compared. The best performing monomers were the zwitterion [2-(methacryloyloxy)ethyl]dimethyl-(3 sulfopropyl)ammonium hydroxide, and diacetone acrylamide, a neutral monomer containing an amide group. Other excellent surfaces were synthesized from amides, amines, basic monomers, and long-chain poly(ethylene) glycols. Bench-scale studies conducted for selected monomers verified the scalability of HTP-PGP results. The results and the synthesis and screening method presented here offer new opportunities for choosing new membrane chemistries that minimize NOM fouling. PMID- 19544901 TI - Reduction of lead oxide (PbO2) and release of Pb(II) in mixtures of natural organic matter, free chlorine and monochloramine. AB - The primary focus of this paper is to elucidate the influence of mixtures of natural organic matter (NOM) and free chlorine and NOM and monochloramine on the reduction of PbO2 in drinking water. Parallel experiments using PbO2 particles of two different sizes (approximately 20 and approximately 200 nm) were conducted to explore the effects of particle size on this process. In the absence of NOM, reduction of PbO2 was observed in monochloramine solutions but not in free chlorine solutions. In the presence of NOM, significant Pb(II) formation was observed in disinfectant-free solutions. The release of Pb(II) was suppressed by the additional presence of free chlorine until the point in time when free chlorine was exhausted. Monochloramine also repressed Pb(II) formation in the presence of NOM but not as significantly as free chlorine. The presence of NOM and monochloramine does not necessarily act additively or synergistically due to complex interactions including reduction of PbO2 by NOM, monochloramine mediated reduction of PbO2, and oxidation of NOM by monochloramine. Higher surface area normalized Pb(ll) formation was found in experiments using larger PbO2 particles. The high reactivity generally associated with nanoparticles was not observed in our study. PMID- 19544902 TI - Assessing the performance of biological filtration as pretreatment to low pressure membranes for drinking water. AB - Although the use of ultrafiltration membranes in drinking water treatment is increasing, fouling remains a major challenge. The objective of this study was to evaluate rapid biological filtration (without coagulant addition) as a pretreatmentto reduce fouling. Surface water was first passed through a pilot scale roughing filter followed by two parallel anthracite/sand biofilters having different contact times, before being fed to the ultrafiltration membrane. As a chemical-free pretreatment, this novel application of biofiltration removes biopolymers (polysaccharides and proteins) that are the most important component of organic matter for fouling, as well as removing particulate matter. Biopolymer removal was influenced by contact time and temperature. The biofilter with the longer contact time led to greater reductions in both hydraulically reversible and irreversible fouling. The extent of hydraulically reversible fouling was related to the membrane influent biopolymer concentration, but the level of hydraulically irreversible fouling was not, indicating that the composition of the biopolymer fraction may have been important. Biofiltration as a simple and robust pretreatment may be particularly suited for small drinking water systems. PMID- 19544903 TI - Greenhouse gas emissions embodied in reinforced concrete and timber railway sleepers. AB - In Australia, there are currently two main materials used for railway sleepers: timber (river red gum, a species of eucalypt) and reinforced concrete. Within the state of Victoria alone, there are currently seven million railway sleepers that make up the rail network. It is estimated that around two million sleepers, or 29%, are presently required to replace timber sleepers and upgrade the entire network, for which there are significant environmental implications, such as the emission of greenhouse gases. These emissions are mainly as a result of the energy and other resources required or "embodied" through the sleeper manufacture, including those associated with harvesting timber and mining raw materials for manufacturing cement. Where alternatives are readily available, it is important that the environmental impacts of the various choices are assessed, ensuring that these impacts are minimized. This study aimed to assess the life cycle greenhouse gas emissions associated with timber and reinforced concrete railway sleepers and showed that the life cycle emissions of reinforced concrete sleepers were up to six times less than the emissions associated with timber sleepers. Taking the potential errors associated with this type of assessment into account, there appearsto be a significant advantage in using reinforced concrete sleepers, in terms of reducing the life cycle emissions associated with the provision of railway sleepers. PMID- 19544904 TI - Partitioning-based dosing: an approach to include bioavailability in the effect directed analysis of contaminated sediment samples. AB - One of the major challenges of effect-directed identification of hazardous chemicals in sediments is the appropriate consideration of bioavailability for a hazard-based prioritization of fractions and toxicants. The equilibrium partitioning approach is frequently used to estimate the bioavailability of sediment contaminants. To simulate partitioning ofthe constituents of complex sediment contaminations in the sediment-water-biota system, a new dosing technique based on silicone elastomer stirrer bars (silicone rods) for use in a growth inhibition test with the green algae Scenedesmus vacuolatus was developed. The dosing technique has a high loading capacity and a fast achievement of equilibrium (5-50 min) for a broad range of model compounds. The subsequent delivery of compounds stored by the silicone during the bioassay compensates possible losses of compounds and keeps the concentrations constant. Furthermore, a dose-dependent response is achieved for single compounds and complex mixtures from sediment extracts. The extracts of three sediment samples from the Elbe River Basin were fractionated using a multistep HPLC method and investigated in the bioassay using conventional DMSO dosing and loaded silicone rods. The results of the toxicity test clearly show that the toxicity patterns are quite different for the two dosing techniques, whereas the differences between the sampling locations are less pronounced. We suggestthat partitioning-based dosing better reflects exposure in sediments and thus provides a more realistic identification of key toxicants. PMID- 19544906 TI - Oxidative potential of semi-volatile and non volatile particulate matter (PM) from heavy-duty vehicles retrofitted with emission control technologies. AB - Advanced exhaust after-treatment devices for diesel vehicles are less effective in controlling semivolatile species than the refractory PM fractions. This study investigated the oxidative potential (OP) of PM from vehicles with six retrofitted technologies (vanadium and zeolite based selective catalytic reduction (V-SCRT, Z-SCRT), Continuously regenerating technology (CRT), catalyzed DPX filter, catalyzed continuously regenerating trap (CCRT), and uncatalyzed Horizon filter) in comparison to a "baseline" vehicle (without any control device). Vehicles were tested on a chassis dynamometer atthree driving conditions, i.e., cruise, transient urban dynamometer driving schedule (UDDS), and idle. The consumption rate of dithiothreitol (DTT), one of the surrogate measures of OP, was determined for PM samples collected at ambient and elevated temperatures (thermally denuded of semivolatile species). Control devices reduced the OP expressed per vehicle distance traveled by 60-98%. The oxidative potential per unit mass of PM however, was highest for the Horizon followed by CRT, DPX Idle, SCRTs, and baseline vehicles. Significant reduction in OP (by 50-100%) was observed forthermally denuded PM from vehicles with retrofitted technologies (PM with significant semivolatile fraction), whereas particles emitted bythe baseline vehicle (with insignificant semivolatile fraction) did not demonstrate any measurable changes in oxidative activity. This suggests that the semivolatile fraction of particles are far more oxidative in nature than refractory particles a conclusion further supported by previous tunnel and ambient studies, demonstrating a decline in PM oxidative activity with increasing atmospheric dilution. Correlation analysis performed between all the species, showed that OP is moderately associated (R = 0.76) with organic carbon (OC) and strongly associated (R = 0.94) with the water-soluble organic carbon (WSOC). PMID- 19544905 TI - Environmental health impacts of equine estrogens derived from hormone replacement therapy. AB - Many factors have been considered in evaluations of the risk-benefit balance of hormone replacement therapy (HRT), used for treating menopausal symptoms in women, but not its potential risks for the environment We investigated the possible environmental health implications of conjugated equine estrogens (CEEs), the most common components of HRT, including their discharge into the environment, their uptake, potency, and ability to induce biological effects in wildlife. Influents and effluents from four U.K. sewage treatment works (STWs), and bile of effluent-exposed fish, were screened for six equine estrogens. In vitro estrogen receptor (ER) activation assays were applied in humans and fish to compare their potencies, followed by in vivo exposures of fish to equine estrogens and evaluation of bioaccumulation, estrogenic responses, and ER gene expression. The equine estrogen equilenin (Eqn), and its metabolite 17beta dihydroequilenin (17beta-Eqn), were detected by tandem GC-MSMS in all STW influent samples and 83% of STW effluent samples analyzed, respectively, at low concentrations (0.07-2.6 ng/L) and were taken-up into effluent-exposed fish. As occurs in humans, these estrogens bound to and activated the fish ERs, with potencies at ERalpha 2.4-3490% of thatfor 17beta-estradiol. Exposure of fish for 21 days to Eqn and 17beta-Eqn induced estrogenic responses including hepatic growth and vitellogenin production at concentrations as low as 0.6-4.2 ng/L. Associated with these effects were inductions of hepatic ERalpha and ERbeta1 gene expression, suggesting ER-mediated mechanism(s) of action. These data provide evidence for the discharge of equine estrogens from HRT into the aquatic environment and highlight a strong likelihood that these compounds contribute to feminization in exposed wildlife. PMID- 19544907 TI - Evaluation of the physiology of Medaka as a model animal for standardized toxicity tests of chemicals by using mRNA expression profiling. AB - The fish acute toxicity test in the Organisation for Economic CoOperation and Development "OECD Guidelines for Testing of Chemicals" is an essential test for environmental toxicity. Here, we have tried to evaluate the physiology of medaka during these test procedures by using genomics. Genomics technology can provide genome-wide expression profiles of mRNA, and these profiles correspond to the physiology of organisms. Thus, a comparison of mRNA expression profiles gives information on the reproducibility of experimental conditions. Expression profiles of mRNA were measured for medakas maintained within the allowable range of the test conditions and also under extreme conditions beyond the guideline limits. We confirmed the high physiological reproducibility of medaka kept in the recommended conditions of the fish acute toxicity test in the "OECD Guidelines for Testing of Chemicals" from the expression profiles obtained under all experimental conditions, except for the types of feeding. PMID- 19544908 TI - How might selenium moderate the toxic effects of mercury in stream fish of the western U.S.? AB - The ability of selenium (Se) to moderate mercury (Hg) toxicity is well established in the literature. Mercury exposures that might otherwise produce toxic effects are counteracted by Se, particularly when Se:Hg molar ratios approach or exceed 1. We analyzed whole body Se and Hg concentrations in 468 fish representing 40 species from 137 sites across 12 western U.S. states. The fish samples were evaluated relative to a published wildlife protective Hg threshold (0.1 sg Hg x g(-1) wet wt.), the currenttissue based methylmercury (MeHg) water quality criterion (WQC) for the protection of humans (0.3 microg Hg x g(-1) wet wt) and to presumed protections against Hg toxicity when Se:Hg molar ratios are >1. A large proportion (56%) of our total fish sample exceeded the wildlife Hg threshold, whereas a smaller, but significant proportion (12%), exceeded the MeHg WQC. However, 97.5% of the total fish sample contained more Se than Hg (molar ratio >1) leaving only 2.5% with Se: Hg ratios <1. All but one of the fish with Se:Hg <1, were of the genus Ptychochelius (pikeminnow). Scientific literature on Se counteracting Hg toxicity and our finding that 97.5% of the freshwater fish in our survey have sufficient Se to potentially protect them and their consumers against Hg toxicity suggests that Se in fish tissue (Se:Hg molar ratio) must be considered when assessing the potential toxic effects of Hg. PMID- 19544909 TI - Classification of chemicals based on concentration-dependent toxicological data using ToxClust. AB - Concentration-dependent response relationships provide essential information on the characteristics of chemical-induced effects on toxicological end points, which include effect (inhibition or induction), potency, and efficacy of the chemical. Recent developments in systems biology and high throughputtechnologies have allowed simultaneous examination of many chemicals at multiple end point levels. While this increase in the quantity of information generated offers great potential, it also poses a significant challenge to environmental scientists to efficiently manage and interpret these large data sets. Here we present a novel method, ToxClust, that allows clustering of chemicals on the basis of concentration-response data derived with single or multiple end points. This method utilizes a least distance-searching algorithm (LDSA) to measure the pattern dissimilarity of concentration-response curves between chemicals and their relative toxic potency. ToxClust was tested using simulated data and chemical test data collected from the human H295R cell-based in vitro steroidogenesis assay. ToxClust effectively identified similar patterns of simulated data and responses to the exposure with the five model chemicals and separated them into different groups on the basis of their dissimilarities. These observations demonstrate that ToxClust not only provides an effective data analysis and visualization tool, but also has value in hypothesis generation and mechanism-based chemical classification. PMID- 19544910 TI - Ecotoxicity of silver nanoparticles on the soil nematode Caenorhabditis elegans using functional ecotoxicogenomics. AB - In the present study, the ecotoxicity of silver nanoparticles (AgNPs) was investigated in Caenorhabditis elegans using survival, growth, and reproduction, as the ecotoxicological endpoints, as well as stress response gene expression. Whole genome microarray was used to screen global changes in C. elegans transcription profiles after AgNPs exposure, followed by quantitative analysis of selected genes. The integration of gene expression with organism and population level endpoints was investigated using C. elegans functional genomics tools, to test the ecotoxicological relevance of AgNPs-induced gene expression. AgNPs exerted considerable toxicity in C. elegans, most clearly as dramatically decreased reproduction potential. Increased expression of the superoxide dismutases-3 (sod-3) and abnormal dauer formation protein (daf-12) genes with 0.1 and 0.5 mg/L of AgNPs exposures occurred concurrently with significant decreases in reproduction ability. Overall results of functional genomic studies using mutant analyses suggested that the sod-3 and daf-12 gene expressions may have been related to the AgNPs-induced reproductive failure in C. elegans and that oxidative stress may have been an important mechanism in AgNPs toxicity. PMID- 19544911 TI - Uranium speciation in drinking water from drilled wells in southern Finland and its potential links to health effects. AB - Exceptionally high concentrations of natural uranium have been found in drinking water originating from drilled wells in Southern Finland. However, no clear clinical symptoms have been observed among the exposed population. Hence a question arose as to whether uranium speciation could be one reason for the lack of significant adverse health effects. Uranium species were determined using time resolved laser-induced fluorescence spectroscopy. We performed multi-element chemical analyses in these water samples, and predictive calculations were carried out using up-to-date thermodynamic data. The results indicated good agreement between measurements and modeling. The low toxicity of Finnish bedrockwater may be due to the predominance of two calcium-dependent species, Ca2UO2(CO3)3(aq) and CaUO2(CO3)3(2-), whose nontoxicity for cells has been described previously. This interdisciplinary study describes chemical speciation of drinking water with elevated uranium concentrations and the potential consequence on health. From these results, it appears that modeling could be used for a better understanding of uranium toxicity of drinking water in the event of contamination. PMID- 19544912 TI - CO2 reaction with hydrated class H well cement under geologic sequestration conditions: effects of flyash admixtures. AB - The rate and mechanism of reaction of pozzolan-amended Class H cement exposed to both supercritical CO2 and CO2-saturated brine were determined under geologic sequestration conditions to assess the potential impact of cement degradation in existing, wells on CO2 storage integrity. The pozzolan additive chosen, Type F flyash, is the most common additive used in cements for well sealing in oil-gas field operations. The 35:65 and 65:35 (v/v) pozzolan-cement blends were exposed to supercritical CO2 and CO2-saturated brine and underwent cement carbonation. Extrapolation of the carbonation rate for the 35:65 case suggests a penetration depth of 170-180 mm for both the CO2-saturated brine and supercritical CO2 after 30 years. Despite alteration in both pozzolan systems, the reacted cement remained relatively impermeable to fluid flow after exposure to brine solution saturated with CO2, with values well below the American Petroleum Institute recommended maximum well cement permeability of 200 microD. Analyses of 50: 50 pozzolan-cement cores from a production well in a sandstone reservoir exhibited carbonation and low permeability to brine solution saturated with CO2, which are consistent with our laboratory findings. PMID- 19544913 TI - Direct biological conversion of electrical current into methane by electromethanogenesis. AB - New sustainable methods are needed to produce renewable energy carriers that can be stored and used for transportation, heating, or chemical production. Here we demonstrate that methane can directly be produced using a biocathode containing methanogens in electrochemical systems (abiotic anode) or microbial electrolysis cells (MECs; biotic anode) by a process called electromethanogenesis. At a set potential of less than -0.7 V (vs Ag/AgCl), carbon dioxide was reduced to methane using a two-chamber electrochemical reactor containing an abiotic anode, a biocathode, and no precious metal catalysts. At -1.0 V, the current capture efficiency was 96%. Electrochemical measurements made using linear sweep voltammetry showed that the biocathode substantially increased current densities compared to a plain carbon cathode where only small amounts of hydrogen gas could be produced. Both increased current densities and very small hydrogen production rates by a plain cathode therefore support a mechanism of methane production directly from current and not from hydrogen gas. The biocathode was dominated by a single Archaeon, Methanobacterium palustre. When a current was generated by an exoelectrogenic biofilm on the anode growing on acetate in a single-chamber MEC, methane was produced at an overall energy efficiency of 80% (electrical energy and substrate heat of combustion). These results show that electromethanogenesis can be used to convert electrical current produced from renewable energy sources (such as wind, solar, or biomass) into a biofuel (methane) as well as serving as a method for the capture of carbon dioxide. PMID- 19544914 TI - Design and testing of an independently controlled urea SCR retrofit system for the reduction of NOx emissions from marine diesels. AB - Diesel engine emissions for on-road, stationary and marine applications are regulated in the United States via standards set by the Environmental Protection Agency (EPA). A major component of diesel exhaust that is difficult to reduce is nitrogen oxides (NOx). Selective catalytic reduction (SCR) has been in use for many years for stationary applications, including external combustion boilers, and is promising for NOx abatement as a retrofit for mobile applications where diesel compression ignition engines are used. The research presented in this paper is the first phase of a program focused on the reduction of NOx by use of a stand-alone urea injection system, applicable to marine diesel engines typical of work boats (e.g., tugs). Most current urea SCR systems communicate with engine controls to predict NOx emissions based on signals such as torque and engine speed, however many marine engines in use still employ mechanical injection technology and lack electronic communication abilities. The system developed and discussed in this paper controls NOx emissions independentof engine operating parameters and measures NOx and exhaust flow using the following exhaust sensor inputs: absolute pressure, differential pressure, temperature, and NOx concentration. These sensor inputs were integrated into an independent controller and open loop architecture to estimate the necessary amount of urea needed, and the controller uses pulse width modulation (PWM) to power an automotive fuel injector for airless urea delivery. The system was tested in a transient test cell on a 350 hp engine certified at 4 g/bhp-hr of NOx, with a goal of reducing the engine out NOx levels by 50%. NOx reduction capabilities of 41-67% were shown on the non road transient cycle (NRTC) and ICOMIA E5 steady state cycles with system optimization during testing to minimize the dilute ammonia slip to cycle averages of 5-7 ppm. The goal of 50% reduction of NOx can be achieved dependent upon cycle. Further research with control optimization, urea distribution and possible use of oxidation catalysts is recommended to improve the NOx reduction capabilities while minimizing ammonia slip. PMID- 19544915 TI - Comment on "Oxidative degradation of organic compounds using zero-valent iron in the presence of natural organic matter serving as an electron shuttle". PMID- 19544916 TI - Comment on "Sustainability of uranium mining and milling: toward quantifying resources and eco-efficiency". PMID- 19544917 TI - Comment on "Growing rice aerobically markedly decreases arsenic accumulation". PMID- 19544918 TI - Correspondence on Geisz et al. Melting glaciers: a probable source of DDT to the Antarctic marine ecosystem. PMID- 19544919 TI - New insight into the oxidation of arsenite by the reaction of zerovalent iron and oxygen. Comment on "pH dependence of Fenton reagent generation and As(III) oxidation and removal by corrosion of zero valent iron in aerated water". PMID- 19544920 TI - Comment on "Food-miles and the relative climate impacts of food choices in the United States". PMID- 19544921 TI - Child abuse, human rights and the role of the doctor. PMID- 19544922 TI - Euthanasia in Belgium five years after legalisation. AB - In 2002, Belgium became the second country to legalise euthanasia after the Netherlands. Three biannual reports have been published by the Federal Control and Evaluation Commission, the body which monitors the application of the law. This article explores how the Belgian law works and what is known about Belgian euthanasia practice both before and since legalisation. PMID- 19544924 TI - Advance directives: binding or merely indicative? Incoherence of the Portuguese National Council of Ethics for the Life Sciences and insufficiencies of newly proposed regulation. AB - The Portuguese National Council of Ethics for the Life Sciences issued in 2005 two important Opinions concerning persistent vegetative state (PVS) and refusal of blood transfusions. The first one advocated that advance directives should be respected; however, the second Opinion considered them "merely indicative." The different opinions of the National Council of Ethics reflect the difficulty of this matter. Portugal ratified the Convention on Human Rights and Biomedicine, which states that advance directives "should be taken into consideration" (Art. 9) and in order to regulate this generic rule, the Portuguese Association on Bioethics proposed to the Parliament a draft-law, which aims to legalize advance directives (including "living will" and "health-care proxy") and establish a National Registry of Advance Directives. This proposal dearly states that advance directives should be binding. However, some regulatory aspects, concerning the procedure that leads to the validity of a living will deserve further discussion. The Author argues in favour of a previous medical interview and a solemn formality in the case of binding advance directives, in order to assure the freedom and information of the refusal of treatment. PMID- 19544923 TI - End-of-life decisions: a view from Ontario and beyond. AB - This article is a comparative study of the law dealing with substitute decision making for incapable patients in end-of-life situations across Canadian provinces. In recent years, this issue has received growing consideration from judicial authorities in Canada and abroad. The article discusses the most significant decisions in Canada, with reference as well to leading cases in the UK, Germany and the US. Future policy and legislative options are discussed as well. PMID- 19544925 TI - Ovarian tissue cryopreservation: promises and uncertainties. AB - Cancer in children and young adults is increasingly being cured by operations, radiotherapy and/or chemotherapy. However, one of the serious side effects of these treatments is the risk of damage to fertility. Whereas the most important goal used to be survival, now increasing attention is being paid to the quality of life in the long term, thanks to the success of these treatments. Infertility affects the quality of life. In post-pubescent boys and men semen can be frozen for later use prior to treatment that harms the spermatogenesis. In girls and young women the solution for reduced fertility or infertility after ovary damaging treatment, may consist of the cryopreservation of ovarian tissue prior to this treatment. At a later stage a decision can be made to transplant this ovarian tissue into the patient or to follow an IVF procedure. There are important normative questions regarding this experimental treatment. The main question is, whether it may be introduced in health care as a regular treatment or should be subject to medical research first. In the Netherlands, a working party of both doctors and ethical, legal and psychological experts recommended to carry out proper research before introducing ovarian tissue cryopreservation in regular health care. This article is meant to elucidate this policy and, including some relevant updates, thus to contribute to the discussion on this question in other European countries. PMID- 19544926 TI - ECHR 2009/5 Case of Juppala v. Finland, 2 December 2008, no. 18620/03 (Fourth Section). PMID- 19544927 TI - ECHR 2009/6 Case of Frankowicz v. Poland, 16 December 2008, no. 53025/99 (Fourth Section). PMID- 19544928 TI - ECHR 2009/7 Case of Aleksanyan v. Russia, 22 December 2008, no. 46468/06 (First Section). PMID- 19544929 TI - ECHR 2009/8 Case of Slawomir Musial v. Poland, 20 January 2009, no. 28300/06 (Fourth Section). PMID- 19544930 TI - ECJ 2009/02 Hecht-Pharma GmbH v. Staatliches Gewerbeaufsichtsamt Luneburg and Vertreterin des Bundesinteresses beim Bundesverwaltungsgericht, 15 January 2009 (C-140/07). PMID- 19544931 TI - Pay-for-performance in nursing homes. AB - Information on the impact of pay-for-performance programs is lacking in the nursing home setting. This literature review (1980-2007) identified 13 prior examples of pay-for-performance programs in the nursing home setting: 7 programs were active as of 2007, while 6 had been terminated. The programs were mostly short-lived, varied considerably in the choice of performance measures and pay incentives, and evaluations of the impact were rare. PMID- 19544932 TI - Payment reduction and Medicare private fee-for-service plans. AB - Medicare private fee-for-service (PFFS) plans are paid like other Medicare Advantage (MA) plans but are exempt from many MA requirements. Recently, Congress set average payments well above the costs of traditional fee-for-service (FFS) Medicare, inducing dramatic increases in PFFS plan enrollment. This has significant implications for Medicare's budget, provoking calls for policy change. We predict the effect of proposals to cut PFFS payments on PFFS plan participation and enrollment. We find that small reductions in payment rates would reduce PFFS participation and enrollment; if Congress reduces payments to traditional FFS levels it would cause the vast majority (85 percent) of PFFS plans to exit the market. PMID- 19544933 TI - Medicaid consumers and informed decisionmaking. AB - In 2006, Florida's Medicaid reform required some Medicaid consumers to enroll in health plans that differed in terms of cost-sharing requirements and benefit limitations. In focus groups we found enthusiasm among Medicaid consumers for having choices among health plans; however, enthusiasm did not translate into comparison shopping for health plans. Survey findings suggested that Medicaid consumers had difficulty comprehending Medicaid health-plan comparison information, particularly if they were lower in numeracy or literacy skills. Given the number of plans offered and the numerous ways they differed, our efforts to simplify the comparison chart resulted in slightly higher comprehension, but only among those with higher skill levels. Our study suggests that policymakers should seek to simplify Medicaid Program information and design to encourage informed decisionmaking. PMID- 19544935 TI - Developing financial benchmarks for critical access hospitals. AB - This study developed and applied benchmarks for five indicators included in the CAH Financial Indicators Report, an annual, hospital-specific report distributed to all critical access hospitals (CAHs). An online survey of Chief Executive Officers and Chief Financial Officers was used to establish benchmarks. Indicator values for 2004, 2005, and 2006 were calculated for 421 CAHs and hospital performance was compared to the benchmarks. Although many hospitals performed better than benchmark on one indicator in 1 year, very few performed better than benchmark on all five indicators in all 3 years. The probability of performing better than benchmark differed among peer groups. PMID- 19544934 TI - Psychometric properties of an instrument to assess Medicare beneficiaries' prescription drug plan experiences. AB - Using data from 335,249 Medicare beneficiaries who responded to the 2007 Medicare Consumer Assessment of Healthcare Providers and Systems (CAHPS) survey, along with data from 22 cognitive interviews, we investigated the reliability and validity of an instrument designed to assess beneficiaries' experiences with their prescription drug plans. Composite measures derived from the instrument had acceptable internal consistency and sufficient plan-level reliability to inform consumer choice, quality improvement, and payor oversight. These measures were positively associated with members' overall rating of the plan and their willingness to recommend the plan. Moreover, each was independently useful in predicting beneficiaries' global ratings of their plan. This instrument can be an important tool for helping beneficiaries to choose a plan that best meets their needs. PMID- 19544936 TI - Use and knowledge of the new enrollee "welcome to Medicare" physical examination benefit. AB - The Medicare Current Beneficiary Survey (MCBS) is a large survey utilizing a nationally representative sample of the Medicare population. The MCBS collects data on a whole host of topics including health status, health insurance coverage and financing, access to care, knowledge and understanding of the Medicare Program, as well as use and effectiveness of new program benefits and changes. PMID- 19544937 TI - Medicare financial status, budget impact, and sustainability--which concept is which? AB - Medicare is continually undergoing change, as it must in order to reflect advances in medical technology, new health care delivery systems, financial pressures, and other developments. Modifications to the program are debated by policymakers in Congress and the administration, together with academic experts and others. These debates would be improved if policymakers and the public had a clearer understanding of Medicare and certain commonly cited views of the program's overall status. Three such concepts--the financial status of the Medicare trust funds, the impact of Medicare on the Federal budget, and the long run sustainability of Medicare-are often confused with each other and are sometimes used interchangeably. Each concept is important but needs to be used for its own purpose. This article clarifies the differences among these three views of Medicare and provides examples of each. PMID- 19544938 TI - Dynamometer measurements of grip and knee extension strength: are they indicative of overall limb and trunk muscle strength? AB - Single dynamometric measures are sometimes used to characterize overall muscle strength. This study examined the legitimacy of that practice by assessing how dynamometer measures of grip and knee extension strength related to manual muscle test grades of each upper and lower limb and of the trunk. Based on correlations and principal component analysis, grip dynamometry can be used to characterize upper limb strength but not lower limb strength; knee extension dynamometry can be used to characterize lower limb and trunk strength. PMID- 19544939 TI - Relation of body mass index and body fat mass for Spanish university students, taking into account leisure-time physical activity. AB - The objective of this cross-sectional study was to analyze the relation of Body Mass Index with body fat mass while taking into account the amount of leisure time physical activity for 299 male university students. Body fat mass was measured by bioelectrical impedance analysis. An estimation of energy expenditure in leisure-time physical activity in metabolic equivalents (METs) was obtained so participants were divided into six activity groups by percentile: no physical activity by the first group and participants physically active were divided into five groups by percentiles: < 25%, 26-50%, 51-75%, 76-90%, and 91-100%. Correlations of Body Mass Index with body fat mass were strong in different groups-values ranged from .76 to .85, except for the > 90% group. PMID- 19544940 TI - Adolescents' cognition of projectile motion: a pilot study. AB - Previous work on the development of intuitive knowledge about projectile motion has shown a dissociation between action knowledge expressed on an action task and conceptual knowledge expressed on a judgment task for young children. The research investigated the generality of dissociation for adolescents. On the action task, participants were asked to swing Ball A of a bifilar pendulum to some height then release it to collide with Ball B, which was projected to hit a target. On the judgment task, participants indicated orally the desired swing angle at which Ball A should be released so that Ball B would strike a target. Unlike previous findings with adults, the adolescents showed conceptual difficulties on the judgment task and well-developed action knowledge on the action task, which suggests dissociation between the two knowledge systems is also present among adolescents. The result further supports the hypothesis that the two knowledge systems follow different developmental trajectories and at different speeds. PMID- 19544941 TI - Measurement of spontaneity: the relationship between intensity and frequency of the spontaneous experience. AB - In the present article, the construct validity of one aspect of the definition of spontaneity is examined. Two ways of measuring spontaneity are presented. One involves the use of the revised version of a Spontaneity Assessment Inventory in which respondents are asked to indicate the intensity of the feeling of spontaneity they experience during a typical day. The second was a specially worded version of the Spontaneity Assessment Inventory-Revised: respondents were asked to indicate the frequency with which they experienced spontaneity during a typical day. 81 students responded to the two versions, which were given 1 wk. apart. The order of administration of the two versions was counterbalanced across the participants. The very high positive correlation between versions (r = .84) suggests that either measure may be used as an estimator of spontaneity. PMID- 19544942 TI - Goal priming and self-efficacy: independent contributions to motor performance. AB - It is well-established that goal pursuit and performance are responsive to the goal characteristics (e.g., difficulty, specificity) and beliefs about self and the task (e.g., self-efficacy). Also, contextual factors may lead to nonconscious goal activation and pursuit. Nevertheless, much remains to be discoverd concerning the way conscious and nonconscious goals are related. In this study, the way in which self-efficacy and nonconscious goal priming may affect a goal directed activity was explored. 67 right-handed participants with high or low self-efficacy on their motor skills performed a drawing task associated with an accuracy instruction. Before task performance, they were primed with accuracy related words (goal-compatible condition), inaccuracy-related words (goal incompatible condition), or received no priming manipulation (control condition). Analysis indicated that performance of the motor task was independently influenced by self-efficacy and goal priming. PMID- 19544943 TI - Effects of prior use of chopsticks on two different types of dexterity tests: Moving Beans Test and Purdue Pegboard. AB - The Purdue Pegboard and Moving Beans with Tweezers test have been used in the rehabilitation of persons with nervous system disorders; however, these two tests differ in their methodology. In the latter test, the testee picks up items with chopsticks or tweezers, but in the former test, the testee grasps items directly with the fingers of one hand. Use experience of a tool may strongly influence performance. The present study examined the use of chopsticks in daily life on performance of the two tests of finger dexterity by two groups of 20 youths who habitually used chopsticks to eat and 20 who did not. Three 30-sec. trials with the dominant and nondominant hands were given. Analysis of variance gave a main effect for number of beans moved between the two groups, hand (nondominant vs dominant), and trials. Significantly more beans were moved by the group with prior chopsticks use than the nonusers by the dominant hand than the nondominant hand, and on Trials 2 and 3 than Trial 1. For the Purdue Pegboard, the only significant difference for trials showed fewer pegs moved on Trial 1 than Trial 3. In conclusion, the groups who habitually used chopsticks performed better on the Moving Beans with Tweezers test than the group without such experience. Also, the marked laterality and practice effects for chopstick users was not observed on the Purdue Pegboard. PMID- 19544944 TI - Role of exercise stages in self-reported health-promoting behaviors of a group of Turkish adolescents at transition to university. AB - The purpose of this study was to examine the self-perceived health-promoting behaviors with respect to exercise stages of Turkish adolescents at transition from high school to university. Participants were 358 adolescents (192 boys, 166 girls) who were accepted to a research university for undergraduate education. The Adolescent Health Promotion Scale was used to examine Nutrition, Health Responsibility, Life Appreciation, Social Support, Exercise and Stress Management behaviors. Physical Activity Stages of Change Questionnaire was used to examine the exercise stages of change. Scores were lowest for Exercise and highest for Life Appreciation regardless of exercise stage. All health-promoting behaviors differed by exercise stage. Participants at lower exercise stages had lower scores. Interventions should focus on increasing physical activity of physically inactive adolescents for holistic development of all health-promoting behaviors. PMID- 19544945 TI - Orientation anisotropy in the Ternus phenomenon. AB - The Ternus display is an ambiguous, multi-element, apparent-movement animation which can be perceived in two alternative organizations. One of these involves the simultaneous movement of several elements and is called group movement. The other involves the movement of one element only and is called element movement. The present study examined the influence of retinal eccentricity and organizational orientation on the percentage of group-movement reports obtained across a range of interstimulus intervals. Six participants observed 48 variations of the Ternus display over 100 replications and judged the type of movement seen on each trial. Analysis showed that retinal eccentricity had no effect on the appearance of the display. However, compared with horizontally organized elements, obliquely organized and vertically organized elements yielded progressively more group movement across all but the shortest interstimulus intervals. These results represent an orientational anisotropy in the processing of Ternus apparent movement. PMID- 19544946 TI - Exercise stages of change in Turkish university students by sex, residence, and department. AB - To examine the exercise behavior of 953 Turkish university students (496 men, 457 women) by sex, residence, and department of study, a stratified sampling method, based on the total number of students in each department, was used to select participants. The mean age of the participants was 21.3 yr. (SD = 1.9). For completed Physical Activity Stages of Change Questionnaire and the International Physical Activity Questionnaire, descriptive statistics and chi-squared tests indicated that most students were at the lower stages of change in exercise habits: Precontemplation (15.2%), Contemplation (31.4%), and Preparation (25.3%), compared with the later stages of Action (7.5%) and Maintenance (20.6%). Women and students from the Department of Architecture were at lower stages of change than men and students from other departments (p < .05), but there were no significant differences between students who were living on or off campus. Stage based interventions would be appropriate, with focus on the physical activity of women and students in architecture. PMID- 19544947 TI - Facilitating effect of natural frequencies: size does not matter. AB - The question of whether humans are able to work in a Bayesian way is currently a topic of substantial investigation. An important finding, reported by Gigerenzer and Hoffrage in 1995 is that Bayesian reasoning is facilitated when the information format corresponds to natural frequencies. The present concern was whether the facilitating effect of frequencies persists when natural frequencies relate to samples which are not convenient multiples of 10. 150 undergraduates participated as volunteers (42 men, 108 women; M age = 23 yr.). Analysis showed the effect of natural frequency formats was not dependent on size of reference class. Theoretical and practical implications are discussed. PMID- 19544948 TI - Resampling permutation probability values for Cronbach's alpha. AB - A permutation procedure to compute a resampling probability value and empirical quantile limits is provided for Cronbach's alpha measure of internal reliability. The underlying assumptions for the conventional use of alpha are discussed. The use of a permutation test for Cronbach's alpha is highlighted as a valuable tool when sample sizes are small and necessary assumptions cannot be met. PMID- 19544949 TI - Effects of playing video games on pain response during a cold pressor task. AB - Two studies assessed whether playing video games would significantly distract participants from painful stimulation via a cold pressor test. In Study 1, participants (8 men, 22 women, M age = 18.5 yr., SD = 1.3) in an action-oriented game condition tolerated pain for a longer time period and reported lower pain intensity ratings than those in a nonaction-oriented game or a nongame control condition. No differences were found on scores of aggressiveness, competitiveness, or prior video game experience, suggesting that these factors play little role. In Study 2, participants (14 men, 13 women, M age = 19.7 yr., SD = 1.3) engaged in six video game conditions (action, fighting, puzzle, sports, arcade, and boxing) and a nongame control condition. Video game play produced an increase in pulse, which was greatest during the action, fighting, sports, and boxing games. Pain tolerance was greatest during the sports and fighting games. Thus, certain games produce greater distraction, which may have implications for the medical field as an adjunct to pain management. PMID- 19544950 TI - Effects of dichotically enhanced electronic communication on crash risk and performance during simulated driving. AB - In a driving simulator experiment, male and female college students received turn by-turn driving directions and cognitive tasks while interacting with an experimenter via an audio communication system. In an Enhanced condition (n = 29), lower speech frequencies (containing the speech fundamental frequency) were routed to participants' left ears (with right cerebral-hemisphere processing) and verbal frequencies above the speech fundamental frequency were routed to right ears (with left hemisphere processing). A control group (n = 31) heard unfiltered communications in both ears. Compared to those in the Control condition, participants in the Enhanced condition were significantly less likely to crash and had nonsignificantly lower rates of driving errors (speed infractions, improper lane position, and following distance errors). The results suggest a means of alleviating cognitive load pressure and mitigating crash risk when complex equipment is operated concurrently with two-way electronic communications (cell phone communication while driving, air-to-air and air-to-ground communications, etc.). PMID- 19544951 TI - Environmental characteristics and physical activity in racial/ethnic minority and Euro-American college students. AB - Little is known about the relations of environmental characteristics and physical activity of college students, especially students from diverse racial or ethnic backgrounds. Associations were examined between environmental characteristics and moderate and vigorous activity in racial or ethnic minority (n = 418; M age = 24.3 +/- 4.5 yr.; 54.2% women) and Euro-American (n = 297; M age = 23.5 +/- 4.4 yr.; 49.7% women) college students. Participants completed questionnaires assessing demographic measures, physical activity, exercise equipment at home, neighborhood characteristics, and convenient places for physical activity. Moderate and vigorous activity participation was similar between the two groups even though racial or ethnic minorities had less supportive home and neighborhood environments for activity and fewer convenient facilities. Greater moderate and vigorous activity were related with more convenient facilities in racial or ethnic minorities. Vigorous activity was associated with better neighborhood characteristics for Euro-Americans. The findings can inform the design and implementation of environmental approaches to promoting activity of college students from diverse ethnic backgrounds. PMID- 19544952 TI - Physical activity and physical self-concept in a sample of middle-age Basque adults. AB - This study examined the relation of physical activity and physical self-concept in a sample of midlife adults from the Basque Country, 248 participants (152 women and 96 men) whose ages ranged from 31 to 49 years. Measurements were made on the Autokontzeptu Fisikoaren Itaunketa, a measure of physical self-concept in Basque language, and questions about physical activity. The Student t test was used to examine differences between Active and Non-active people; and to analyse differences by frequency of activity, analysis of variance was used. Analysis indicated those women who identified themselves as more active had more positive self-perceptions with regard to physical ability, physical condition, strength, and physical self-concept than inactive people while men's scores were more positive for physical ability, physical condition, and physical self-concept. Likewise, both men and women who reported engaging in physical activity more than three times a week also had significantly more positive self-perceptions with regard to physical ability, physical condition, and physical self-concept than those who exercised less frequently. PMID- 19544953 TI - Comparison of expert and nonexpert swimmers' opinions about the value, potency, and activity of four standard swimming strokes and underwater undulatory swimming. AB - Underwater undulatory swimming (UUS) is often perceived to be a nonessential aspect of aquatic propulsion. Given their solid theoretical and practical training in swimming, physical education students should be capable of judging the true value of the "fifth stroke," since it appears to be the most efficient technique in high level, competitive swimming. To compare opinions and connotations associated with the stroke and the four official strokes (butterfly, backstroke, breaststroke, and crawl), 198 students (32 of whom were expert swimmers; M age = 20.6 yr., SD = 1.2), were surveyed using the semantic differential of Osgood, Suci, and Tannenbaum. Although answers of expert and nonexpert swimmers differed significantly (p < .01, except for the breaststroke), participants considered overall that undulatory stroke was less attractive, less powerful, and less rapid than the four surface strokes (d = 2.88 for the expert swimmers). Putting one arm in front of the other and repeating the sequence still remains the most solidly held representation of "the right way" to swim. However, the high observed standard deviations for the underwater undulatory stimulus (SD > or = 1.1 with SD max = 3 for the expert swimmers) attests to the view being less strongly held by swimming specialists. PMID- 19544954 TI - Correlates of learning in introductory biomechanics. AB - Characteristics hypothesized to be related to learning in introductory biomechanics classes were examined. Data from a 2003 national study were supplemented by additional instructor-reported data about experience, tests, laboratory hours, and average course-related expenditures. The number of credit hours of the course (r = .15) and money spent by departments on laboratory classes per year (r = -.18) were significantly and uniquely correlated with learning. Increasing credit hours from 3 to 4 with laboratory experience doubled learning. The results supported national standards for a 4-credit course and an emphasis on conceptual understanding rather than quantitative problem-solving. Researchers should seek to confirm these results as well as explore whether contact hours or the nature of laboratory learning experiences contribute to this effect. PMID- 19544955 TI - Learning and context of use for small-screen leading displays on visual performance in a Chinese sample. AB - Leading displays represent mechanisms for exhibiting temporal instead of spatial information to overcome the limited display space of mobile devices. Prior studies focused only on information presented but disregarded the influence of context of use and learnability. In this study, 12 Chinese-speaking college students were presented a small-screen mobile device that simultaneously showed 100 Chinese characters and a 30-character leading display. Analyzed were presentation rate (250, 350, and 450 characters per minute), presentation mode (character-by-character or word-by-word), and learning (5 practice sessions) by instructing the subjects to perform a static information search task to identify the location of target characters on the screen and through an evaluation of reading comprehension for the text presented in the leading display. There was no significant change in performance over the 5 days of practice, but the rate of presentation and word-by-word presentation significantly affected reading comprehension. Results indicated that none of the leading-display factors distracted subjects from the static information-search task, but they were influenced by comprehension of the leading-display content. PMID- 19544956 TI - Children's imitations of movements are goal-directed and context-specific. AB - Previous research indicates that imitation of gestures in preschool children is goal-directed. A goal may be a salient feature from a presented movement; that goal may be imitated correctly, but other features were ignored, resulting in observable errors. Objects (e.g., a dot on the table) can become the most salient features and presence or absence of objects influences imitation responses. Imitation responses were examined under conditions in which objects could not be used directly as the most salient feature. 60 children (M age = 5:6) were assigned to Gestural, Dot, No-dot, and Un-dot conditions, and they were asked to imitate 20 movements. The type of presented movement and the occurrence of correct, mirror, and error responses were examined. Responses in the Un-dot condition were similar to those in the Dot condition. Error responses in the Un dot condition were related to age. Children may extract a more abstract feature in a context without visible objects. This ability is associated with a cognitive mechanism developed in preschool years. PMID- 19544957 TI - A study of optimal handle shape and muscle strength distribution on lower arm when holding a foil. AB - The strength of five working muscle groups of the lower arms of 8 male fencers, including adductor pollicis, extensor carpi radialis, flexor carpi radialis, extensor carpi ulnaris, and flexor carpi ulnaris, were examined during competition. Root mean square values of muscular electromyographic signals indicated that the shape of foil handles significantly influenced distribution of working strength of each muscle group. Use of the Pistol-Viscounti type of foil handle showed better distribution of strength among the 5 muscle groups than did other types of foils. Using the Pistol-Viscounti foil handle not only reduced muscular fatigue but also lessened cumulative trauma symptoms while holding a foil for a long duration. PMID- 19544958 TI - Influence of attention to somatic information on emotional and autonomic responses. AB - The present study aimed to investigate the dissociable effects of two forms of self-focus on emotional and autonomic responses. One form is suppression, which includes the suppression of heart rate and self-evaluation of performance. The other is observation, which includes attention to one's own heart rate with no suppression and no evaluation. 26 undergraduate and graduate students from the Nagoya University campus (13 men, 13 women), ages 18 to 24 years (M = 20.7, SD = 1.6) were recruited. Participants were provided with their own heart rate as feedback for 5 min., during which participants conducted a self-focus manipulation. Several days after the experimental session for one condition, the same participants conducted another experimental session for the other condition. Instruction to suppress enhanced physiological arousal and subsequent negative emotions; however, instruction to observe did not increase physiological arousal or negative emotions. PMID- 19544959 TI - Verbal regulation of grip force in preschoolers. AB - The purpose of this study was to clarify the developmental processes in verbal regulation by preschool children. Participants were 152 typically developing children (74 boys, 78 girls) between 4 and 6 years of age (M = 5.3, SD = .8), and 30 healthy adults (15 men, 15 women) between 19 and 26 years of age (M = 20.8, SD = 1.4). In Exp. 1, the task was to regulate grip force based on quantitative instruction which implies using a scale for regulation. Participants were required to produce a half-grip force of the maximum (Task 1). In Exp. 2, the task was grip-force regulation based on nonquantitative instruction. The participants were asked to respond with a slightly weaker grip force than the maximum (Task 2) and then a further weaker grip force (Task 3) than that used on Task 2. The regulation rates produced the extent of regulation and suggest regulation by quantitative instruction may develop earlier than by nonquantitative instruction. Also, precise grip-force regulation based on the semantic aspect of instruction may be difficult for young children. The developmental changes in the rate of performance especially observed in children of 4 to 6 years indicate that the tendency to use too much grip force disappears during this preschool period. In addition, too little grip force in regulation may reflect the developmental process toward fine grasping movements. PMID- 19544960 TI - Relation of physical activity and self-esteem. AB - The aim of this study was to examine the relation between self-esteem appraisal and physical activity testing a convenience sample of 211 individuals, ages 19 to 35 years and selected from the general population after a brief structured interview. They were grouped by sport habits into three distinct groups named Athletes, Nonathletes, and Sedentary people, and then were examined for significant differences in self-esteem scores measured via the Heatherton and Polivy State Self-esteem Scale which assesses three correlated factors, respectively, Performance, Social, and Appearance. As hypothesized, self-esteem scores between-groups differences emerged for the Appearance factor only, and the Sedentary group scored comparatively lower than the other two groups. PMID- 19544961 TI - Interaction of hand preference with eye dominance on accuracy in archery. AB - In this study, the interaction of hand preference, eye dominance, and performance in archery was investigated. Beginners' accuracy (right-handed French students (48 men, 34 women) from the University of Sport Sciences (M age = 19.3 yr., SD = 1.7 yr.), were tested and 1,323 practitioners were given a laterality questionnaire. Analysis suggested the interaction of hand preference and eye dominance might influence accuracy of the novice archers (uncrossed patterns were more accurate) when the bow was used without sights, but the use of sights by practitioners seemed to eliminate this effect. PMID- 19544962 TI - Insincere utterances and gaze: eye contact during sarcastic statements. AB - Anecdotal evidence suggests that speakers often gaze away from their listeners during sarcastic utterances; however, this question has not been directly addressed empirically. This study systematically compared gaze-direction of speakers in dyadic conversation when uttering sincere and sarcastic statements. 18 naive participants were required to recite a series of contradictory statements on a single topic to a naive listener, while at the same time conveying their actual opinion about this topic. This latter task could only be accomplished through prosodic or nonverbal communication by indicating sincerity or insincerity (sarcasm) for the various statements and allowed examination of gaze across the two conditions for each participant. Subsequent analysis of the videotaped interaction indicated that, during the time for the actual utterance, sarcastic utterances were accompanied by greater gaze aversion than were sincere utterances. This effect occurred for 15 of 18 participants (3 men, 15 women; M age = 19.8, SD = 1.0) who had volunteered for a small credit in an Introductory Psychology course. Results are discussed in terms of nonverbal communication and possible miscommunication which may apply given cultural differences in use of nonverbal cues. PMID- 19544963 TI - Selection of resistance exercise intensity using ratings of perceived exertion from the OMNI-RES. AB - This investigation evaluated the use of OMNI-Resistance Exercise Scale ratings of perceived exertion as a method for selecting training intensities that are appropriate for improving muscular fitness. 20 women (M = 21.3 yr., SD = 2.2) completed a one-repetition maximum for the chest press and knee extension. Participants returned for three sessions during which they were asked to select resistances which elicited the feelings of exertion associated with ratings of 3 (i.e., Easy to Somewhat easy), 6 (i.e., Somewhat hard) and 9 (i.e., Hard to Extremely hard). The average percentage of maximum lifted at a 3, 6, and 9 was 50, 69, and 88% for the chest press and 56, 74, and 90% for the knee extension, respectively. Effect sizes indicated large differences across the three ratings of perceived exertion within sessions and small differences between sessions. Intraclass correlation coefficients ranged from .69 to .95. The results indicated that OMNI-Resistance Exercise Scale ratings of 3, 6, and 9 can be used to accurately and reliably select intensities that are appropriate for improving muscular fitness. PMID- 19544964 TI - Regional differences in spatial frame of reference systems for people in different areas of China. AB - Levinson defined spatial frame of reference systems into three types and found that people who spoke different languages all over the world had different habits in the use of these systems. Some quasi-experimental designed research in China suggested that Chinese people in various areas had different preferences for frame of reference systems in nonlinguistic spatial tasks. In this study, a newly designed and strictly controlled laboratory experiment was conducted to measure performance on linguistic spatial tasks. Response times of two groups of participants, respectively selected from the north (10 men and 12 women; M age = 24 yr., SD = 4) and south (11 men and 12 women; M age = 24 yr., SD = 3) of China, were compared on processing of spatial terms used in different systems: absolute (e.g., east) versus relative (e.g., right). To reduce the effect of living experience, the Northern participants with less than 4 yr. living experience in the south of China were included, and vice versa. Analysis showed that Southerners, but not Northerners, differed in response times to terms between absolute and relative systems. This finding suggests that speed of processing spatial terms differs among people who speak the uniform language Mandarin but live in geographically distinct home areas in China and use different frame of reference systems. PMID- 19544965 TI - Music and light during indoor cycling. AB - The present research is a field study assessing effects of music and light on physical performance and subjective feelings of 28 participants (14 men, 14 women) in fitness classes using indoor cycling. Participants performed four sessions under different conditions of music and light. Analysis showed a sense of pleasure was significantly higher when music was introduced during the exercise. A significant interaction of music and light indicated that participants experienced less sense of tiredness when they trained with music and lights were dimmed. Light alone had no effect on any of the subjective measures. Physiological measurements showed that light and music did not influence amount of energy exerted (heart rate) or energy expended (calories). Apparently, participants do not work harder when music is present; however, they report more pleasure and less tiredness while exercising with music and dimmed light. It is recommended that music and dimmed light might be used during such activities to heighten pleasure and reduce tiredness while emphasizing fitness achieved. PMID- 19544966 TI - Postural stability of athletes in Special Olympics. AB - To assess body equilibrium in athletes with intellectual disability, 60 adults with intellectual disability (30 Down syndrome, 30 nonsyndromic) participating in the 2005 Italian Special Olympics games were tested, and data for 30 healthy control adults were tested. Each subject performed four posturographic tests with open eyes, open eyes and cotton rolls between antagonist teeth, and closed eyes and cotton rolls between teeth. For each subject and test, oscillations of the body's center of foot pressure on a force platform were measured. Comparisons of the center of foot pressure sway area between groups were computed. Ratios of the sway area for the center of foot pressure among experimental conditions were compared for all athletes. The athletes with Down syndrome had larger sway of center of foot pressure area than controls and smaller than that of athletes for the nonsyndromic. All participants oscillated less with open eyes than with closed eyes. The cotton rolls reduced the sway area for the center of foot pressure by participants, while athletes with intellectual disability showed larger body sway than healthy ones, but cotton rolls between the teeth seemed to improve their postural performance. PMID- 19544967 TI - Perception of approaching and retreating floor-projected shapes in a large, immersive, multimedia learning environment. AB - Perception of floor-projected moving geometric shapes was examined in the context of the Situated Multimedia Arts Learning Laboratory (SMALLab), an immersive, mixed-reality learning environment. As predicted, the projected destinations of shapes which retreated in depth (proximal origin) were judged significantly less accurately than those that approached (distal origin). Participants maintained similar magnitudes of error throughout the session, and no effect of practice was observed. Shape perception in an immersive multimedia environment is comparable to the real world. One may conclude that systematic exploration of basic psychological phenomena in novel mediated environments is integral to an understanding of human behavior in novel human-computer interaction architectures. PMID- 19544968 TI - Is manipulation of color effective in study of the global precedence effect? AB - This article evaluates the use of color manipulation in studying the effect of global precedence and the possible involvement of the magnocellular processing system. The analysis shows variations of color used in three studies produced changes on the global precedence effect, but findings based on this technique present some methodological problems and have little theoretical support from the magnocellular processing-system perspective. For this reason, more research is required to develop knowledge about the origin of these variations in global precedence. PMID- 19544969 TI - Effects of colored lights, spacing between stimuli, and viewing distance on error in a depth-matching task. AB - A previous study indicated that color of a comparison stimulus, colored light, and illumination significantly affected the error in perceived depth with binocular and monocular vision. Here, the effects of spacing between standard and comparison stimuli, viewing distance, and five colored lights were investigated. 32 Taiwanese university students in five groups were assigned to one of five colored light conditions. Spacing between comparison and standard stimuli significantly affected error in a depth-matching task. Errors increased with increased spacing. Colored light also had a significant effect; error rate in the depth-matching task under the green light condition was significantly greater than the rates under the white, yellow, and red light conditions, but the difference in error rates between blue and green and among the white, yellow, and red light conditions were not significant. The error rate at a viewing distance of 110 cm was significantly less than errors under 70- and 150-cm conditions. In addition, a two-way interaction effect was detected: viewing distance with spacing between standard and comparison stimuli. PMID- 19544970 TI - Bone marrow-derived progenitor cells could modulate pancreatic cancer tumorigenesis via peritumoral microenvironment in a rat model. AB - Metaplastic tubular complexes (MTC) have been proposed as precursor lesions for pancreatic adenocarcinoma (PDAC). In this study, we investigated the potential role of bone marrow-derived progenitor cells (BMPC) in the formation of MTC and PDAC in a rat model. F344 rats defective for CD26 (dipeptidyl peptidase IV, DPPIV) expression were sublethally irradiated and received rescue bone marrow cells from wild-type F344 rats that express CD26. After confirming engraftment, recipient animals received dimethylbenzanthracene (DMBA) implantation in their pancreas. Animals were sacrificed monthly from 3 to 7 months. We observed both MTC and tumors in animals that received DMBA. These MTC were ductal complexes because they stained positive for cytokeratin but were negative for chymotrypsin and chromogranin A. Cells that expressed both CD26 and cytokeratin were rarely observed in the MTC. Cells expressing either both CD26 and CD45 or CD26 and smooth muscle actin were also found near the MTC. However, no CD26 signal was detected in the tumors. Within this model, there appeared to be no evidence supporting that BMPC turned into tumor cells directly. BMPC could modulate pancreatic cancer growth through tumor microenvironment. PMID- 19544971 TI - COX-2 polymorphisms in patients with familial adenomatous polyposis. AB - Cyclooxygenase-2 (COX-2) is an enzyme involved in the synthesis of prostaglandins and thromboxanes, which are regulators of biologic processes such as inflammation, cell proliferation, and angiogenesis. COX-2 has been found overexpressed in (pre)malignant tissues and may be relevant to cancer development. We investigated whether functional genetic polymorphisms in COX-2 may have a risk-modifying effect on duodenal adenomatosis in patients with familial adenomatous polyposis (FAP). Blood from 85 patients with FAP and 218 age and sex-matched healthy subjects was investigated for the presence of two functional promoter region polymorphisms (-1195G-->A and -765G-->C) in COX-2. Logistic regression analysis revealed an overrepresentation of the -1195GG genotype compared to the -1195AA genotype in patients with FAP (odds ratio = 2.81; 95% CI = 1.00-7.91, p = 0.042). No associations between single COX-2 polymorphisms or COX-2 haplotype were found when patients were evaluated according to their Spigelman stage. The predicted low COX-2 expression genotype 1195GG was found overrepresented in the patients with FAP. The COX-2 genotypes showed no association with the severity of duodenal adenomatosis. PMID- 19544972 TI - Identification of endogenous reference genes for qRT-PCR analysis in normal matched breast tumor tissues. AB - Quantitative gene expression measurements from tumor tissue are frequently compared with matched normal and/or adjacent tumor tissue expression for diagnostic marker gene selection as well as assessment of the degree of transcriptional deregulation in cancer. Selection of an appropriate reference gene (RG) or an RG panel, which varies depending on cancer type, molecular subtypes, and the normal tissues used for interindividual calibration, is crucial for the accurate quantification of gene expression. Several RG panels have been suggested in breast cancer for making comparisons among tumor subtypes, cell lines, and benign/malignant tumors. In this study, expression patterns of 15 widely used endogenous RGs (ACTB, TBP, GAPDH, SDHA, HPRT, HMBS, B2M, PPIA, GUSB, YWHAZ2, PGK1, RPLP0, PUM1, MRPL19, and RPL41), and three candidate genes that were selected through analysis of two independent microarray datasets (IL22RA1, TC22, ZNF224) were determined in 23 primary breast tumors and their matched normal tissues using qRT-PCR. Additionally, 18S rRNA, ACTB, and SDHA were tested using randomly primed cDNAs from 13 breast tumor pairs to assess the rRNA/mRNA ratio. The tumors exhibited significantly lower rRNA/mRNA ratio when compared to their normals, on average. The expression of the studied RGs in breast tumors did not exhibit differences in terms of grade, ER, or PR status. The stability of RGs was examined based on two different statistical models, namely GeNorm and NormFinder. Among the 18 tested endogenous reference genes, ACTB and SDHA were identified as the most suitable reference genes for the normalization of qRT-PCR data in the analysis of normal matched tumor breast tissue pairs by both programs. In addition, the expression of the gelsolin (GSN) gene, a well-known downregulated target in breast tumors, was analyzed using the two most suitable genes and different RG combinations to validate their effectiveness as a normalization factor (NF). The GSN expression of the tumors used in this study was significantly lower than that of normals showing the effectivity of using ACTB and SDHA as suitable RGs in this set of tumor-normal tissue panel. The combinational use of the best performing two RGs (ACTB and SDHA) as a normalization factor can be recommended to minimize sample variability and to increase the accuracy and resolution of gene expression normalization in tumor normal paired breast cancer qRT-PCR studies. PMID- 19544973 TI - Breast cancer risk associated with polymorphisms of IL-1RN and IL-4 gene in Indian women. AB - Interleukins and cytokines are important regulator of the aetio-pathogenesis of the majority of cancers. Mechanistic role of IL-1RN and IL-4, particularly in breast carcinogenesis, is well documented. However, the role of polymorphisms of IL-1RN and IL-4 combinations associated with risk of breast cancer is not reported. The IL-1RN and IL-4 gene polymorphisms were genotyped with VNTR-PCR in 100 patients (benign tumor n = 32 and breast cancer n = 68) and 200 normal healthy control subjects with normal mammogram. Genotype distribution and allelic frequencies between patients and controls were compared and odds ratios (OR) with 95% confidence intervals (CI) were calculated using SPSS software (version 12.0). There were no significant differences in the genotype distributions of both IL 1RN and IL-4 polymorphisms between cases and controls. Similarly, subgroup analysis showed that there is no significant association for pre- and postmenopausal women. However, BB genotype of IL-1RN significantly differs among benign and malignant stages of breast cancer. IL-1RN and IL-4 polymorphisms alone or in combination are not associated with risk of breast cancer in Indian patients. The association of IL-1RN with malignant stages may indicate its possible role in progression of breast cancer. Further studies in other population are needed to confirm our findings and to elucidate the role of IL-1RN in progression of breast cancer. PMID- 19544974 TI - Sodium selenite induces apoptosis in acute promyelocytic leukemia-derived NB4 cells through mitochondria-dependent pathway. AB - Our previous study has shown that sodium selenite can cause apoptosis in acute promyelocytic leukemia-derived NB4 cells in a caspase-dependent manner involving Deltapsim disruption and cleavage of Bcl-2, but more detailed mechanism(s) remain unclear. Here we showed that mitochondrial apoptosis signaling pathway played a vital role in apoptosis induced by sodium selenite based on the following findings: 1) cytochrome c release, activation of caspase 9, mitochondrial targeting, and oligermerization of Bax; 2) caspase 9, but not caspase 8, inhibitor could attenuate apoptosis; 3) downregulation of Bax and Bad by siRNA could delay sodium selenite-induced apoptosis. Further investigation showed that ROS was an essential inducer of deltapsim disruption and apoptosis by sodium selenite. Our findings here demonstrate that sodium selenite can induce apoptosis in NB4 cells through a mechanism involving ROS, activation of proapoptotic proteins Bad and Bax, Deltapsim disruption, release of cytochrome c, and consequent initiation of caspase cascade. PMID- 19544976 TI - Widen recruitment net to avoid crisis. PMID- 19544975 TI - Endothelin-1 inhibition by ambrisentan as a potential treatment adjunct after debulking surgery in epithelial ovarian cancer. AB - The 21 amino acid signaling peptide endothelin-1 is commonly elevated in epithelial ovarian cancer, and it mediates or facilitates much of this cancer's aggressive behavior. Ambrisentan (Letairis; Gilead Sciences Inc.) is an antagonist of endothelin-1 at its cognate receptor that has just been approved to treat pulmonary hypertension. Ambrisentan is a well-tolerated pill taken once daily. In theory, it should retard and inhibit lodgement and establishment of disseminated peritoneal micrometastases after debulking surgery. PMID- 19544977 TI - An end to violence? PMID- 19544978 TI - Current ward manager roles do not reflect nurses' career ambitions. PMID- 19544980 TI - Nurses' role in helping to rehabilitate patients to return to health after critical illness. PMID- 19544979 TI - Developing community-based activities for inpatients in a mental health hospital. AB - A regional mental health hospital with a successful hospital-based activity programme created an activity coordinator role. The aim was to promote social inclusion by encouraging patients to take part in activities in mainstream settings. This article outlines the process of developing the activity programme. Patients' feedback was extremely positive. PMID- 19544981 TI - Exploring patients' experiences of a nurse-led follow-up service after critical care. AB - AIM: To explore the impact of critical care experiences on patients' long-term health. METHOD: This was a mixed-method interview and questionnaire study that involved patients in the design. Patients were interviewed at different times up to 12 months after critical care discharge. RESULTS: Thirty-seven interviews were carried out with 28 patients. Themes included: rehabilitation from critical care; memories; uncertainty and fear; and empathy; with reassurance as a core theme. Psychological implications need to be considered and nurses' sensitivity and preventative care can help patients deal with the stress and aftermath of critical care. CONCLUSION: Follow-up services, such as the nurse-led service described here, are important in supporting patients. PMID- 19544982 TI - New guidance on how to avoid inserting female-length urinary catheters into men. PMID- 19544983 TI - Effective team leadership: techniques that nurses can use to improve teamworking. AB - This is the second article in a two-part series on leadership. The first examined transformational approaches to effective leadership in healthcare settings. This article describes the strategies that nurses can use to ensure healthcare teams are effective: team reflection; ensuring all members participate; and establishing ground rules. PMID- 19544984 TI - Health promotion in sexual health 2: how to put theory into practice and empower clients. AB - This is the second in a two-part unit on health promotion in sexual health. Part 1 outlined various theories and models on the issue. This part examines the factors that contribute towards successful health promotion, such as an effective communication style. It outlines how nurses can put health-promotion theory, competencies and guidance into practice. PMID- 19544985 TI - On how technology can help nurses improve patient care. PMID- 19544986 TI - 'The NHS has a moral duty not to waste its resources'. PMID- 19544987 TI - [Quantifying direct N2O emissions from paddy fields during rice growing season in China: model and input data validation]. AB - The models on direct N2O emissions from rice paddies under different water regimes developed by the authors were validated against field measurements in China reported in 2005-2007 and in other regions. In flooding rice paddies (F), N2O emission predicted by the model was consistent with previous reports in other regions. Under the water regime of flooding-midseason drainage-reflooding (F-D F), the model developed in this study was comparable to that established by using worldwide database. The models also well fitted N2O emissions from rice paddies under the water regime of flooding-midseason drainage-reflooding-moisture but without waterlogging (F-D-F-M) in China. Consistency of rice production data derived from the database of this study with those reported in previous studies suggests that the model input data of rice production had high reliability. The input data showed that water management and nitrogen input regimes have greatly changed in rice paddies since the 1950s. During the 1950s-1970s, about 20%-25% of the rice paddy was continuous water logging, and 75%-80% under the water regime of F-D-F. Since the 1980s, about 12%-16%, 77% and 7%-12% of paddy fields were under the water regimes of F, F-D-F and F-D-F-M, respectively. Total N input during the rice growing season averaged 87.49 kg x hm(-2) in the 1950s and 224.64 kg x hm(-2) in the 1990s. Chemical N input during the rice growing season has increased from 37.4 kg x hm(-2) in the 1950s to 198.8 kg x hm(-2) in the 1990s, accounting for 43% and 88% of the seasonal total N inputs, respectively. Manure N input was applied at stable rate, ranging from 45.2 kg x hm(-2) to 48.2 kg x hm( 2) during the 1950s-1970s, but thereafter it decreased over time. The contribution of manure N to total N inputs has decreased from 52% in the 1950s to 9% in the 1990s. Crop residue N retained during the rice growing season has increased from 4.9 kg x hm(-2) in the 1950s to 6.3 kg x hm(-2) in the 1980s. A high spatial variation of nitrogen inputs during the rice growing season was found in the 1950s-1970s, while it was not pronounced in the 1980s-1990s. Overall, the results of this study suggest that the models could be used to quantify direct N2O emissions from rice paddies under various water regimes in China. PMID- 19544988 TI - [Quantifying direct N2O emissions from paddy fields during rice growing season in China: model application]. AB - Based on statistical analysis of field N2O measurements in rice paddies in China, the models on direct N2O emissions under different water regimes were established. After successes in model validation and input data verification, the models were used to quantify changes in direct N2O emissions from paddy fields during the rice growing season in mainland China between the 1950s and the 1990s. Due to increases in rice planting area and nitrogen input and changes in water regime, the models predicted that seasonal N2O-N emissions from rice paddies have increased from 9.55 Gg each year in the 1950s to 32.26 Gg N2O-N in the 1990s, which was accompanied by the increase in rice yield over the period 1950s-1990s. During the period 1950s-1990s, seasonal N2O-N emissions from rice paddies have increased, on average, at a rate of 6.74 Gg per decade. Seasonal N2O fluxes in rice paddies were estimated to be 0.32 kg x hm(-2) in the 1950s and 1.00 kg x hm( 2) in the 1990s, which accounted for 0.37% and 0.46% of the seasonal total N inputs, respectively. The uncertainties in N2O estimate were estimated to be 59.8% in the 1950s and 37.5% in the 1990s. Seasonal N2O emissions from rice paddies in the region of middle and lower Yangtze River contributed 51% -56% to its national total. In the 1990s, N2O emissions during the rice growing season accounted for 8%-11% of the reported annual total of N2O emissions from croplands in China, suggesting that paddy rice development could have contributed to mitigating agricultural N2O emissions in the past decades. However, seasonal N2O emissions would be increased given that saving-water irrigation and nitrogen inputs are increasingly adopted in rice paddies in China. PMID- 19544989 TI - [Inventories of atmospheric arsenic emissions from coal combustion in China, 2005]. AB - Anthropogenic arsenic (As) emitted from coal combustion is one of key trace elements leading to negative air pollution and national economy loss. It is of great significance to estimate the atmospheric arsenic emission for proposing relevant laws or regulations and selecting proper pollution control technologies. The inventories of atmospheric arsenic emissions from coal combustion in China were evaluated by adopting the emission factor method based on fuel consumption. Arsenic emission sources were firstly classified into several categories by economic sectors, combustion types and pollution control technologies. Then, according to provincial coal consumption and averaged arsenic concentration in the feed fuel, the inventories of atmospheric arsenic emission from coal combustion in China in 2005 were established. Coal outputand consumption in China in 2005 were 2,119.8 and 2,099.8 Mt, respectively. The total emissions of arsenic released into the atmosphere in 2005 in China were estimated at about 1,564.4 t, and Shandong ranked the largest province with 144.4 t arsenic release, followed by Hunan (141.1 t), Hebei (108.5 t), Henan (77.7 t), and Jiangsu (77.0 t), which were mainly concentrated in the eastern and central provinces of China. The arsenic emissions were largely emitted by industry sector (818.8 t) and thermal power generation sector (303.4 t), contributing 52.3% and 19.4% of the totals, respectively. About 375.5 t arsenic was estimated to be released into the atmosphere in the form of gas phase in China in 2005, with a share of 24% of the totals. In general, arsenic pollution control from coal combustion should be highlighted for the power and industry sectors in the whole country. However, arsenic poisoning caused by residential coal burning should also be paid great attention in some areas such as Xinjiang, Gansu, Qinghai and Guishou. PMID- 19544990 TI - [Emission characteristics of fine particles from grate firing boilers]. AB - Grate firing boilers are the main type of Chinese industrial boilers, which accounts for 85% of the industrial boilers and is one of the most important emission sources of primary air pollutants in China. In this study, five boilers in three cities were selected and tested to measure the emission characteristics of PM2.5, and gaseous pollutants were applied by a compact dilution sampling system, which was developed for this field study. Results showed that particles mass size distributions for the five industrial boilers presented single peak or double peak, former peaks near 0.14 microm and the later peaks after 1.0 microm; the cyclone dust remover and wet scrubber dust remover had effective removal efficiencies not only to PM2.5, but also to PM1.0; and under the condition of same control techniques, grate firing boiler with high capacity has less PM2.5 emission than the boiler with low capacity. In the PM2.5 collected from flue gases, SO4(2-) was the most abundant ion, accounted for 20%-40% of the PM2.5; and C was the most abundant element (7.5%-31.8%), followed by S (8.4%-18.7%). Carbon balance method was applied to calculate the emission factors of these pollutants. The emission factors of PM2.5, NO, and SO2 were in the range of 0.046-0.486 g x kg(-1), 1.63-2.47 g x kg(-1), 1.35-9.95 g x kg(-1) respectively. The results are useful for the emission inventory development of industrial boilers and the source analysis of PM2.5 in atmospheric environment. PMID- 19544992 TI - [Contribution of isoprene emitted from vegetable to atmospheric formaldehyde in the ambient air of Beijing city]. AB - Methacrolein (MACR) and methyl vinyl ketone (MVK), which are the characteristic products of isoprene photooxidation in the atmosphere, as well as formaldehyde, were determined during March to November of 2006 in the ambient air of Beijing city. Cartridges coated with 2, 4-dinitrophenylhenylhydrazine (DNPH) were used to collect those carbonyls in ambient air and a high pressure liquid chromatography (HPLC) was used for quantification. Distinct amounts of MACR and MVK were detected during vegetable growing seasons from April to October with ambient levels in ranges of 0.11 x 10(-9) - 0.67 x 10(-9) and 0.19 x 10(-9) - 1.36 x 10( 9), respectively, with their maximal concentrations appearing in August. MACR and MVK were measured under the detecting limit in March and November, which may be ascribed to low isoprene emission from the broadleaf trees without leaves during two months. According to the stoichiometric coefficients among MVK, MACR, isoprene and formaldehyde, the concentrations of the reacted isoprene due to photo-oxidation can be calculated derived from the concentrations of the measured MVK and MACR. Then, the monthly (April to October) average concentrations of formaldehyde produced by the photo-oxidation of reacted isoprene were roughly estimated to be in the range of 0.35 x 10(-9) - 2.5 x 10(-9), which accounted for 4.6%-11.5% of total atmospheric formaldehyde. During the period with active atmospheric photo-oxidation (June to August), the contribution of biogenic isoprene to formaldehyde was predominant. Therefore, the biogenic isoprene is an important source to local photo-oxidant formation in Beijing and should be considered of. PMID- 19544991 TI - [Estimate of the formation potential of secondary organic aerosol in Beijing summertime]. AB - Fractional aerosol coefficients (FAC) are used in conjunction with measurements of volatile organic compounds (VOC) during ozone episodes to estimate the formation potential of secondary organic aerosols (SOA) in the summertime of Beijing. The estimation is based on the actual atmospheric conditions of Beijing, and benzene and isoprene are considered as the precursors of SOA. The results show that 31 out of 70 measured VOC species are SOA precursors, and the total potential SOA formation is predicted to be 8.48 microg/m3, which accounts for 30% of fine organic particle matter. Toluene, xylene, pinene, ethylbenzene and n undecane are the 5 largest contributors to SOA production and account for 20%, 22%, 14%, 9% and 4% of total SOA production, respectively. The anthropogenic aromatic compounds, which yield 76% of the calculated SOA, are the major source of SOA. The biogenic alkenes, alkanes and carbonyls produce 16%, 7% and 1% of SOA formation, respectively. The major components of produced SOA are expected to be aromatic compounds, aliphatic acids, carbonyls and aliphatic nitrates, which contribute to 72%, 14%, 11% and 3% of SOA mass, respectively. The SOA precursors have relatively low atmospheric concentrations and low ozone formation potential. Hence, SOA formation potential of VOC species, in addition to their atmospheric concentrations and ozone formation potential, should be considered in policy making process of VOCs control. PMID- 19544993 TI - [Elemental size distribution of airborne fine and ultrafine particulate matters in the suburb of Shanghai, China]. AB - The elemental size distributions of airborne fine/ultrafine particulate matters in the suburb of Shanghai were studied using synchrotron X-ray fluorescence. Median mass aerodynamic diameter (MMAD), elemental correlation coefficient as well as enrichment factor (EF) of each size fraction were calculated to characterize the sources of elements in fine/ultrafine particulate matters. Ca and Ti distributed mainly in coarse particles (> 2 microm) with size independent enrichment factors between 0.1 and 3.2, and the correlation coefficient between Ca and Ti was as high as 0.933, which implied strong contribution from nature sources, such as soil dusts and resuspended dusts. However, V, Cr, Mn, Ni, Zn, Cu, Pb, Cl, S mainly distributed in 0.1-1.0 microm particulate matters with MMAD between 0.56-0.94 microm. The EF of V, Cr, Ni, Cu, Zn, Pb increased with decreasing particle size. The highest EF were found for Pb in ultrafine particulate matters (< 0.1 microm) with EF of 2,023.7-2,244.2. The evidences suggested that these elements were significantly influenced by anthropogenic sources and enriched in fine/ultrafine particles smaller than 1 microm. Fe distributed uniformly in the particles larger than 0.2 microm with MMAD of 1.3 microm. The results indicated non-negligible influences of remote transmission of anthropogenic pollutions. PMID- 19544994 TI - [Element characteristics and sources of PM2.5 at Mount Dinghu in 2006]. AB - To study the characteristics and sources of particles in Pearl River Delta Region, samples of PM2.5 were collected by high volume sampler, and elements of the samples were determined by ICP-MS. The results showed that concentrations of Pb, V, Cu, As, Zn and Se were 216.24, 15.40, 60.56, 31.81,432.06 and 8.12 ng x m( 3) respectively, those were in high level. Factor analysis on the chemical composition of PM2.5 showed that fuel combustion, metal-working industry, ash and sea salt were the main sources of PM2.5 in this region. PMID- 19544995 TI - [Research on source profile of aerosol organic compounds in leather plant]. AB - Through investigating current air pollution condition for PM10 in every factories of different style leather plants in Pearl River Delta, characteristic profile of semi-volatile organic compounds in PM10 emitted from leather factories and their contents were researched by using ultrasonic and gas chromatography and mass spectrum technology. The 6 types of organic compounds containing 46 species in total were found in the collected samples, including phenyl compounds, alcohols, PAHs, acids, esters and amides. The concentrations of PM10 in leather tanning plant, leather dying plant and man-made leather plant were 678.5, 454.5, 498.6 microgm x m(-3) respectively, and concentration of organic compounds in PM10 were 10.04, 6.89, 14.21 microg x m(-3) in sequence. The more important type of pollutants in each leather plants had higher contribution to total organic mass as follows, esters and amides in tanning plants profile account for 43.47% and 36.51% respectively; esters and alcohols in dying plants profiles account for 52.52% and 16.16% respectively; esters and amide in man-made leather plant have the highest content and account for 57.07% and 24.17% respectively. In the aerosol organic source profiles of tested leather plants, 9-octadecenamide was the abundant important species with the weight of 26.15% in tanning plant, and Bis(2-ethylhexyl) phthalate was up to 44.19% in the dying plant, and Bis(2 ethylhexyl) maleate and 1-hydroxy-piperidine had obviously higher weight in man made plant than the other two plants. PMID- 19544996 TI - [Simultaneous desulfurization and denitrification by TiO2/ACF under different irradiation]. AB - The supported TiO2 photocatalysts were prepared in laboratory, and the experiments of simultaneous desulfurization and denitrification were carried out by self-designed photocatalysis reactor. The optimal experimental conditions were achieved, and the efficiencies of simultaneous desulfurization and denitrification under two different light sources were compared. The results show that the oxygen content of flue gas, reaction temperature, flue gas humidity and irradiation intensity are most essential factors to photocatalysis. For TiO2/ACF, the removal efficiencies of 99.7% for SO2 and 64.3% for NO are obtained respectively at optimal experimental conditions under UV irradiation. For TiO2/ACF, the removal efficiencies of 97.5% for SO2 and 49.6% for NO are achieved respectively at optimal experimental conditions under the visible light irradiation. The results of five times parallel experiments indicate standard deviation S of parallel data is little. The mechanism of removal for SO2 and NO is proposed under two light sources by ion chromatography analysis of the absorption liquid. PMID- 19544997 TI - [Establishment and application of four-dimensional fluxes emission factor model for construction fugitive dust]. AB - We established a four-dimensional fluxes mathematics model and a suit of relevant monitoring project that can calculate construction fugitive dust emission according to actual field monitoring data, this mathematics model is similar to exposure profiling method recommended by U.S. EPA. Four-dimensional fluxes mathematics model is simpler and more convenient to calculate construction fugitive dust emission, and it can preferably eliminate the influence of incidental factors on calculation result. Actual field monitoring data from forty construction sites of Beijing were inputted into the model to calculate TSP emission factor among construction fugitive dust of Beijing local which equaled to 0.492 kg/(m2 x 30 d). By comparing emission factor of Beijing and U.S. EPA's AP-42 documents, we found the ratio between Beijing's and U.S. EPA's emission factor value was 1.83. Beijing local emission factor from this paper is recommended to establish construction fugitive dust emission inventory of Beijing. PMID- 19544998 TI - [Estimation of the optically active substances in case II water of the lower Jinjiang River based on the bio-optical model]. AB - The water's Inherent Optical Properties (IOPs), including absorption and scattering coefficients of water components, are the essential parameters for bio optical model and retrieval of water quality using the semi-analytical method. Nevertheless, the application of the bio-optical model in river water studies is still very rare. Therefore, taking the lower Jinjiang River of Fujian, SE China as an example, this study measured and calculated the bio-optical properties of river water and concentrations of optically active substances based on in situ water samples collected from river in 2007. It shows that R(0(-))753, R(0( ))702/R(0(-))680 and R(0(-))670/R(0(-))423 can be used to estimate total suspended solids (TSS) concentration, phytoplankton pigment (PP) concentration and the CDOM absorption at 440 nm, respectively. The determination coefficients (R2) of the retrieval model of TSS, PP and CDOM are 0.953, 0.8205 and 0.6213, respectively. The corresponding relative errors of the models (RE) are 6.1%, 21.87% and 22.18%. The results show that the model for estimating TSS can achieve the highest accuracy, the PP-estimating model has the second highest accuracy and the CDOM-estimating model has the lowest. The relatively lower concentration of phytoplankton pigments, narrow characterized spectral range of CDOM and influence of CDOM's R(0(-)) by TSS and PP within this spectral range contributed to their relatively lower accuracy. PMID- 19544999 TI - [Numerical simulation of TP transport after overflow of rainwater into urban lake]. AB - Based on the three-dimensional advection-diffusion-reflection equation, a two dimensional TP transport equation was deduced to simulate TP distribution and transport after overflow rainwater into urban lake from storm sewer system during rainstorm. The model has a good agreement with a group of monitor data at Lake Lichee in Shenzhen, China. The model was applied to compute the scenario in Lake Lichee under the design rainstorm, and analyse the fate of TP. It shows that TP flux into lake is 15.385 kg under city storm intensity of 28 mm/h, in which 62.3% of flux goes into water in lake and 28.1% TP flux settles surface sediment. It would take 3.0 days for the integrated treatment project operation to recover TP to the level before the rain. PMID- 19545000 TI - [Algae-removal effect of AS/PDM composite coagulants to winter Taihu Lake raw water]. AB - The series of stable AS/PDM composite coagulants prepared by polydimethyldiallylammonium chloride (PDM) and aluminium sulphate (AS) were used to research the algae-removal effect to winter Taihu Lake raw water. The effects of dosage of composite coagulants, composite mass ratios (20:1-5:1) of AS and PDM, intrinsic viscosity values (0.55-3.99 dL/g) of PDM on algae-removal rates were studied through coagulation and algae-removal experiments. The feasibility of using composite coagulants to substitute prechlorination process was analysed. The results show that when residual turbidity of 2 NTU to water after coagulation and sediment is required by water plant, the dosage (based on Al2O3) of AS, AS/PDM (0.55/20:1-3.99/5:1) composite coagulants are 4.24 mg/L, 3.96-1.87 mg/L, and the algae-removal rates are 83.00%, 87.52%-90.93% respectively. When dosage to raw water are 4.24 mg/L, the algae-removal rates of AS, AS/PDM (0.55/20:1 3.99/5:1) composite coagulants are 83.00%, 88.29%-97.66%, and the residual turbidities are 2.00 NTU, 1.76-0.43 NTU respectively. When dosage to chlorine added water are 4.50 mg/L, the treatment effect of AS/PDM (1.53/10:1) composite coagulant to raw water is better than that of AS to chlorine-added water, and the treatment effect of AS/PDM (3.99/5:1) composite coagulant to raw water is better than that of AS, AS/PDM (0.55/20:1) and AS/PDM (1.53/10:1) composite coagulants to chlorine-added water. So using AS/PDM composite coagulants can enhance evidently the treatment effect of AS to winter Taihu Lake raw water. Compared with using AS solely, the dosage of AS in composite coagulants are saved when the residual turbidities are required in same level, and the treatment effect of AS is enhanced when the dosage of AS in composite coagulants are same as that of using AS solely. Moreover, using composite coagulants can replace the part chlorine-added function on increasing coagulation and algae-removal in prechlorination process and profitably increases safety of drinking water quality. PMID- 19545001 TI - [Effect of hydraulic loading on antioxidant enzyme and digestive enzyme activities of earthworm in biofilter]. AB - Through different working condition experiments, effect of hydraulic loading on antioxidant enzyme (superoxide dismutase, catalase) and digestive enzyme (cellulose, alkaline phosphatase) activities of earthworm in biofilter were studied. The results indicated that antioxidant enzyme and digestive enzyme activities of earthworms responded to hydraulic loading stresses differently. SOD and CAT activities of earthworms increased with the increase of hydraulic loading in the scale of 2.4-6.7 m3 x (m2 x d)(-1), earthworms resisted external environmental stress by coordination function of antioxidation system, so they could survive in different conditions. In addition, digestive enzyme activity, digest ability, sludge reduction and stabilization effect showed excellent correlation (p < 0.05). When the hydraulic loading was 4.8 m3 x (m2 x d)(-1), earthworm biofilter showed a high level of AKP, FP and digest rate (41.47%), which was notably better than that of other conditions, sludge and organic matter reduction rate came to the maximum, 48.2% and 65.5% respectively. However, higher level of hydraulic loading [> or = 6.0 m3 x (m2 x d)(-1)] restrained AKP and FP activity markedly and metabolism level was affected, sludge reduction rate had certain decadence, so it was unfavorable for earthworm biofilter to bring the ecological function into play. At last it is recommended that the hydraulic loading of earthworm biofilter should not exceed 6.0 m3 x (m2 x d)(-1). PMID- 19545003 TI - [Kinetics and degradation efficiency of benzotriazole in water by ozonation]. AB - Benzotriazole (BTri) is widely used in industrial processes and households as anti-icer and corrosion inhibitors. However, the removal of BTri hasn't been reported yet. The results of an experimental study are presented which provide values for the reaction rate constants between molecular ozone and BTri, and overall reaction rate constants for the degradation of BTri in the pH range of 7 8. The kinetic rate constant for BTri degradation by molecular ozone was calculated to be 20.18 L x (mol x s)(-1) according to the experimental data. The study showed that the reaction between ozone and BTri was influenced by some factors. The degradation rate of BTri increased with the increase of the solution pH value and the water temperature, but changed little in the presence of some anions, such as NO3(-), SO4(2-) and Cl(-). The hydroxyl radical inhibitor tert Butyl alcohol affected the removal of BTri to some degree, which proved that the ozone molecule was cooperated with the hydroxyl radical to remove BTri in the oxidation process. PMID- 19545002 TI - [Characteristics and comparative study of a new drinking-water defluoridation adsorbent Bio-F]. AB - To evaluate the application potentiality pf a new type drinking-water defluoridation adsorbent Bio-F, comparative study on the defluoridation characteristics of common adsorbents activated alumina (AA), bone char (BC), activated clinoptilolite (AC) with Bio-F was conducted. The defluoridation characteristics under different conditions, such as particle diameter, pH, retention time, fluorine concentration, regeneration stability, were investigated by continuous-flow column experiments and static tests. The defluoridation efficiency of high fluoride underground water by four types of adsorbents was also compared. The results showed that F(-) adsorption kinetics of Bio-F fitted the Lagergren First-order equation (R2 = 0.9580). F(-) adsorption by Bio-F was found to fit the Langmuir adsorption isotherm (R2 = 0.9992). The results indicated that the static defluoridation capacity (DC) of Bio-F was 4.0883 mg x g(-1), which was about 1.8 folds and 5.8 folds of those of AA and AC respectively. DC of all four adsorbents was positively correlated with F(-) concentration and negatively correlated with particle size. High concentration of CO3(2-) and HCO3(-) reduced the DC of Bio-F (p < 0.05), while high concentration of Ca2+, NO3(-), HPO4(2-) favored defluoridation by Bio-F (p < 0.001). The optimal retention time of Bio-F was 3-4 min, which was less than that of AC (20 min) and AA (11 min). The DC of Bio-F remained relatively stable in pH 4.0-9.0 and in regeneration since the DC variation was not more than 15%. The above results indicated that Bio-F was superior to AA, BC and AC in drinking-water defluoridation. PMID- 19545004 TI - [Modified kinetics model of degradation of MCLR by advanced oxidation process]. AB - Primary extracted MCLR (Microcystin-LR) from Taihu Lake degradation by UV/H2O2 and ozone process were investigated. The effects of intensity of UV radiation, initial concentration of MCLR, dosages of H2O2 and ozone, initial pH value of the solution and anions on degradation of MCLR were studied. The pH value and carbonate (or bicarbonate) concentration varieties during the reaction were discussed. A concise model based on the first order model was raised. The results reveal that in UV/H2O2 system (including UV alone), the average value of the pseudo first and second order kinetics correlation coefficient R2 are 0.927 and 0.827 respectively. In the ozone treatment process, the average valve of the first and second order kinetic correlation coefficient R2 are 0.911 and 0.925 respectively; at the same time, the accuracy of modified model was proved to be better, the average value of R2 in UV/H2O2 (including UV alone) and ozone system are up to 0.982 and 0.985 respectively. Meanwhile, it can be concluded from the model that: when the initial UV radiation, H2O2 dosage and the ozone dosage were set, pH value and carbonate (or bicarbonate) concentrations were the two main aspects which influenced the reaction rate. PMID- 19545005 TI - [Reduction of chromium (VI) by nanoscale zero-valent iron supported on Al pillared bentonite]. AB - In the presence of Al-pillared bentonite with good sorption capacity, nanoscale zero-valent iron supported on Al-pillared bentonite (NZVI/Al-PILC) was prepared with NaBH4 and FeSO4 aqueous solution. The structure of NZVI/Al-PILC was characterized by X-ray diffraction (XRD) and Brunauer-Emmett-Teller (BET). The effects of pH values and initial chromium (VI) concentrations on its removal rate by NZVI/Al-PILC were investigated, and were compared with those of unsupported nanoscale zero-valent iron (NZVI) containing the same iron mount of NZVI/Al-PILC. The results indicate that in the same experimental condition, the chromium (VI) removal by NZVI/Al-PILC reached 100% after 120 min. The removal is not only much higher than that (63.0%) of the NZVI containing same iron mount, but also superior to the sum of removal (75.4%) by NZVI containing the same iron amount and the Al-pillared bentonite containing the same clay amount with NZVI/Al-PILC. PMID- 19545006 TI - [Property analysis of nanosized iron sulfide produced by sulfate reducing bacteria and its application in the treatment of wastewater containing high concentration of Cr (VI)]. AB - The properties of novel nanosized iron sulphide produced by sulfate reducing bacteria (SRB) and its application to the removal of Cr (VI) from wastewater were studied. The structural properties were examined by transmission electron microscope (TEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). The influences of pH, temperature and dosage on removal of Cr (VI) from wastewater by nanosized iron sulphide were investigated, and the effect comparison of nanosized iron sulphide and other traditional wastewater treatment was also evaluated. TEM result shows that the length of the nanosized iron sulphide is approximately 45-80 nm, and the ratio of length to width is 10-15. XPS and XRD results indicate that the Fe/S is 1.07-1.11, and the main composition of nanosized iron sulphide is amorphous FeS and mackinawite. While in treating wastewater containing Cr (V), pH, temperature and dosage of nanosized iron sulphide are the main factors to control the reaction. The Cr (VI) reduction rate increases with decrease of pH and increase of dosage and temperature. Among these factors, pH is the most significant influence factor. At 25 degrees C, pH 3, when the mole ratio of nanosized iron sulphide to chromium (VI) reaches 1.17/1, wastewater containing Cr (VI) 0.03 mol/L can be treated to meet the discharge standard within 10 min. Moreover, in treating wastewater containing Cr (VI), compared with commercial iron sulfide (AR.), nanosized iron sulphide has advantages of low dosage and short treatment time, and compared to conventional biological treatment by SRB, nanosized iron sulphide has advantages of low effluent COD and small amount of sludge. PMID- 19545007 TI - [Phosphorus removal and mechanisms for advanced treatment of sewage by Spirogyra]. AB - The novel way of advanced treatment of municipal wastewater was proposed and characterized by purifying wastewater by Spirogyra. The characteristics of nitrogen and phosphorus removal by Spirogyra were studied aiming at nitrogen and phosphorus removal in advanced treatment of sewage. Under natural light, when Spirogyra's dosage (gross mass) was more than 3.05 g/L, total phosphorus concentration, NH4+-N concentration, TN concentration and permanganate index decreased to less than 0.09, 2.82, 4.31 and 16.86 mg/L respectively, with removal efficiencies of more than 96.84%, 88.60%, 85.49% and 24.56%. During the treatment of sewage, pH value increased, while both calcium cation and magnesium cation concentration decreased, and conductivity decreased. During the growth of Spirogyra, increasing pH value induced saline minerals precipitation, and the precipitated minerals adsorbed phosphate, which were considered as the main mechanisms of phosphorus removal. Furthermore, phosphorus removal by Spirogyra followed Langmuir adsorption isotherm model, and the saturated adsorption of Spirogyra (gross mass) for phosphorus was 3.159 mg/g under this test condition. The increasing pH value made ammonia evaporate during Spirogyra-growth, which might be the main mechanism of nitrogen removal. Under suitable Spirogyra's dosage and hydraulic retention time, the treated sewage might meet the demands of water supply for landscape impoundments on nitrogen and phosphorus concentrations. The better performance of Spirogyra removing nitrogen and phosphorus might provide a novel alternative way for advanced treatment of sewage. PMID- 19545009 TI - [Effects of salinity concentration on N2O production during nitrification]. AB - The experiment investigated the production and conversion rate of N2O during nitrification using saline sewage and municipal wastewater. Different kinds of sludge were used, domesticated by saline sewage influent (salinity 7.5 g/L) and municipal wastewater (salinity 0.1 g/L), respectively. The results showed that the production of N2O using saline sewage was 2.85 times higher than that using municipal wastewater. The production and conversion rate of N2O during nitrification under different salinities were also investigated. The results showed that the production of N2O was almost the same when salinity decreased from 7.5 g/L to 5.0 g/L, even 2.5 g/L. However, specific ammonia oxidation rate was increased with the decrease of salinity. The sudden increase of salinity, from 7.5 g/L to 10 g/L, resulted in the increase of N2O production and conversion rate but decrease of specific ammonia oxidation rate. Consequently, It is important to avoid the severe fluctuation of salinity resulted in the increase of N2O production and conversion rate. PMID- 19545008 TI - [Real-time process control of denitrifying phosphorus and nitrogen removal systems with NO3(-) and NO2(-) as electron acceptors]. AB - The simultaneous phosphorus and nitrogen removal using denitrifying phosphate accumulating organisms with NO3(-) and NO2(-) as electron acceptors was investigated in an anaerobic/anoxic process. The results showed that the concentrations of COD, PO4(3-) and electron acceptors (NO3(-) and NO2(-)) have a correlation with the variation of pH and ORP during anaerobic/anoxic process. In anaerobic phase, the occurrence of pH flat indicates the end of phosphorus release. In anoxic process, an inflexion was observed in ORP profile when the phosphorus uptake was finished. In addition, the phosphorus uptake rate was investigated in the process with different electron acceptors (NO3(-) and NO2( )). The average anoxic phosphate uptake rate was 32.68 mg/(L x h) in the early 30 min during anoxic phase when NO3(-) was as electron acceptor, and every 1 mg PO4(3-)-P uptake would consume 1.14 mg NO3(-)-N. In the process with NO2(-) as electron acceptor, the average anoxic phosphate uptake rate was 17.66 mg/(Lx h) in the early 30 min, and every 1 mg PO4(3-)-P uptake would consume 1.57 mg NO2(-) N. In conclusion, ORP and pH can be used as control parameters for simultaneous phosphorus and nitrogen removal in an anaerobic/anoxic process, and the system with NO3(-) as electron acceptor is superior to the system with NO2(-) as electron acceptor. PMID- 19545010 TI - [Analysis and establishment of control modes of advanced nitrogen removal via nitrite in SBR]. AB - To achieve stable advanced nitrogen removal via nitrite in SBR, factors influencing pH variation in treating real domestic wastewater were investigated. The characteristic points were found to be appeared obviously on pH profile at both ends of ammonia oxidization and denitrification by analyzing the mechanism of biological nitrogen removal and the balance of carbonic acid. Using pH as control parameter in SBR for treating domestic or municipal wastewater with low organic concentration and suitable alkalinity could achieve advanced nitrogen removal with effluent TN below 1 mg/L. Moreover, it could prevent the decrease of nitrite accumulation rate causing by excessive aeration, which plays an important role on the stability of advanced nitrogen removal via nitrite. Based on theoretical analysis and experimental study, the real-time control strategy of advanced nitrogen removal via nitrite was established. To accommodate the different water quality and retain the accuracy of control strategy, 18 adjustable variables were set up in control strategy. The control strategy established a foundation for developing control software and control system of advanced nitrogen removal via nitrite. PMID- 19545011 TI - [Denitrification using radiation-pretreated wheat straw as solid carbon source]. AB - Wheat straw after radiation pretreatment was used as solid carbon source and biofilm support for denitrifying microorganisms. Denitrification performance of radiation-pretreated wheat straw was compared to that of wheat straw without radiation pretreatment. The results showed that the denitrification rate of radiation-pretreated wheat straw was about 20% higher than that of wheat straw without radiation pretreatment. When the initial nitrate concentration was 65.3 mg/L, the denitrification rate using wheat straw after 300 kGy radiation with gamma-ray could reach 0.087 mg/(g x h) and the nitrate removal efficiency was above 90%. Parts of these results were confirmed by the IR analysis and SEM observation of wheat straw surface structure. PMID- 19545012 TI - [Impact of particle on UV disinfection for wastewater]. AB - Water samples with similar particle distribution were irradiated by low-pressure UV. Experiment results were input into the two-kinetic model for calculation. The approach applied in this experiment allowed the assessment of the effect of particles on UV inactivation of FC in their natural state. The results showed that FC concentration increased with the particle number. Particle significantly lowered the inactivation rate of the indigenous FC in the secondary water with elevated particle content when the particle number was less than 10(5) C x mL( 1). On the other hand, the impact was not very significant when the particle number exceeded 10(5) C x mL(-1). Based on the calculation results of a two kinetic model, for the different number of particles, the ratio of the FC which was difficult to inactivate was stable, while most of the FC which were holed in the particles were easy to inactivate. PMID- 19545013 TI - [Analysis of the secondary effluent of municipal wastewater in north China]. AB - The secondary effluent from a municipal sewage treatment plant was fractionated into three parts: the dissolved, the near-colloidal and the suspended. The variation of each composition was investigated individually during winter, spring and summer. The fractionation study revealed that the dissolved COD and DOC accounted for 78.2%-86.5% of total COD and 82.6%-86.6% of TOC, respectively. The molecular weight distribution (MWD) of the secondary effluent indicated that the organics with molecular weight (MW) < 1000 accounted for 56.3%-62.8% of total organics. The organic components constituted 75.54%-89.93 % of the mixture of suspended and near-colloidal matter. The size distribution of the particles showed that the particles ranging from 2.00 to 6.84 microm constituted more than 80% of the total particles. The secondary effluent was relatively poor biodegradable with only 0.195-0.283 of BOD5/COD ratio and 0.156-0.26 of BDOC/DOC ratio. The GC/MS analysis detected 43 and 26 kinds of compounds in dissolved and in suspended forms, respectively, and most of them are bio-refractory according to the functional groups of the chemicals. Biodegradability of the secondary effluent should be enhanced for improving the sequential biochemical removal efficiency for the organic matter. PMID- 19545014 TI - [Gradient elevation of temperature startup experiment of thermophilic ASBR treating thermal-hydrolyzed sewage sludge]. AB - Startup experiment was conducted for thermophilic anaerobic sequencing batch reactor (ASBR) treating thermal-hydrolyzed sewage sludge using the strategy of the step-wise temperature increment: 35 degrees C-->40 degrees C-->47 degrees C- >53 degrees C. The results showed that the first step-increase (from 35 degrees C to 40 degrees C) and final step-increase (from 47 degrees C to 53 degrees C) had only a slight effect on the digestion process. The second step-increase (from 40 degrees C to 47 degrees C) resulted in a severe disturbance: the biogas production, methane content, CODeffluent and microorganism all have strong disturbance. At the steady stage of thermophilic ASBR treating thermal-hydrolyzed sewage sludge, the average daily gas production, methane content, specific methane production (CH4/CODinfluent), TCOD removal rate and SCOD removal rate were 2.038 L/d, 72.0%, 188.8 mL/g, 63.8%, 83.3% respectively. The results of SEM and DGGE indicated that the dominant species are obviously different at early stage and steady stage. PMID- 19545015 TI - [Effect of acid and alkali on phosphorus release from sewage sludge in the microwave treatment process]. AB - Impacts of acid and alkali addition on phosphorus release from sewage sludge were investigated in a microwave reactor at atmospheric pressure. Results showed that in the microwave irradiation process, though NaOH pretreatment increased the release efficiency of phosphorus from 28.8% to 59.1%, large amounts of carbon and nitrogen were correspondingly released into the supernatant. Compared to the alkali pretreatment, pretreatment of H2SO4 not only resulted in the higher release efficiency of phosphorus by 3 times (up to 84.5%), but also sharply reduced the amount of carbon and nitrogen released into the supernatant. These results suggest that H2SO4 pretreatment of sewage sludge can selectively release carbon, nitrogen and phosphorus, which is more suitable for phosphorus recovery from sewage sludge. Meanwhile, the pH range of 2.1-2.2 was optimal for phosphorus release from sewage sludge in the microwave treatment process. PMID- 19545016 TI - [Effect of Ce3+ on volatile fatty acid concentrations during anaerobic granular sludge digestion]. AB - Batch experiments were conducted to investigate the effect of Ce3+ on volatile fatty acid(VFA) concentrations by anaerobic granular sludge digestion using D Glucose and acetic sodium as substrate in the state of stabilization and restart up. Results show that when the concentration of Ce3+ is lower than 1 mg/L, VFA concentration decreases, which suggests the transformation of butyric acid to acetic acid and acetic acid to methane is promoted. When the concentration of Ce3+ is 1-10 mg/L, the bacterial activity decreases and decomposition of the acetic acid and butyric acid becomes more difficult compared with the control. Adding Ce3+ brings little change in the constitution of VFA: 96% of VFA is acetic acid and butyric acid, while the propionic acid accounts for less than 3%. With the acetic sodium as the sole carbon and energy source, adding 0.05 mg/L Ce3+ could accelerate acetate degradation. After being conserved for 4 months, the activity of the Ce-containing anaerobic granular sludge is higher than that of the Ce-free sludge. The present of Ce contributes to the restart-up of anaerobic reactors. PMID- 19545017 TI - [Optimal formation conditions and analytical methods of the target product by MAP precipitation]. AB - In order to establish optimal conditions of the struvite (MAP: MgNH4PO4 x 6H2O) formation, acid dissolution was applied and developed to perform element analyses on the precipitates obtained from MAP precipitation, and a novel analyzing and calculating method was developed to quantitatively determine the struvite content (purity) in the harvested precipitates according to the NH4+-N content. With this method, the purities of struvite were respectively determined for both ultra pure water and tap water used as solutes. At the same time, the effect of pH and Ca2+ on the formation and crystallization of struvite was evaluated. The newly developed method was effective enough to determine the purities of struvite, which could be a better method than qualitative X-ray diffraction (XRD). Based on the developed method, it was found that the optimal pH ranges for having a high struvite content (> 90%) were respectively at 7.5-9.0 with ultra pure water as solute and at 7.0-7.5 with tap water as solute. In real wastewater, Ca2+ at pH > 8.0 might result in impurities rather than struvite. Therefore, a neutral pH range (< 8.0) is proposed to perform struvite precipitation in wastewater. PMID- 19545018 TI - [Polycyclic aromatic hydrocarbons partition in particle-water interface in the Yangtze Estuarine and nearby coastal areas]. AB - Based on polycyclic aromatic hydrocarbons (PAHs) field data of particle phase and dissolved phase, partition coefficients Kp in particle-water interface from the Yangtze Estuarine and nearby coastal areas were obtained. The results show partition coefficients of PAHs in dry season were higher than those in flood season, and all of them ranged from 507 L/kg to 10,179 L/kg, increasing with the rise of PAH compounds. The linear free-energy relationship coefficients between Koc value and octanol-water partition coefficients Ko, in dry season and flood season were 0.82 and 0.68, respectively. These relationships inferred that suspended particle matters in the Yangtze Estuarine and coastal areas had weakly lipophilic characteristics and adsorption ability. lgKoc values of PAH compounds in the different sampling sites were higher than the upper limit value predicted by the classic equilibrium partition model. The partition processes of PAHs were not controlled by concentrations of suspended particle matters, particle grain sizes, overlying water salinity and dissolved organic carbon ( R2 < 0.1), but influenced by particle organic carbon and heterogeneous mixtures of particle soot carbon. The expanded partition model which included soot partitioning could well predict in situ partition processes of PAH compounds whose octanol-water partition coefficients lgKow was lower than 6. PMID- 19545019 TI - [Sorption and desorption of phenanthrene on sediments]. AB - Batch experiments were employed to investigate the influences of salinity on the sorption of phenanthrene. Influences of initial phenanthrene concentration, sediment properties and salinity on desorption of phenanthrene were also studied and the mechanism for desorption hysteresis was deeply discussed. Salinity enhanced the sorption of phenanthrene on sediments, and the enhancement extent was related to sediment organic matter (SOM). A salting constant of 0.125 L x mol(-1) was derived from the PHE solubility in fresh water and salt water, and in the presence of sediment, salting constant was enhanced to 0.299 L x mol(-1). Desorption of phenanthrene all exhibited hysteresis to some extents and desorption hysteresis were described by a thermodynamic index TII. With the initial phenanthrene concentration increasing from 0.5 mg x L(-1) to 2.5 mg x L( 1), the value of TII first decreased from 0.80 to 0.38 and then increased to 0.55, suggesting the desorption hysteresis of phenanthrene first decreased and then increased. Sediment treated by NaOH showed a reduced sequestration ability on phenanthrene than the original sediment did, with the value of TII decreased from 0.55 to 0.33. Desorption of phenanthrene in fresh water was found for the first time to have a greater hysteresis than in salt water, indicated by the greater TII value of 0.55 in fresh water than that of 0.42 in salt water. Desorption hysteresis could be explained by the sorption of phenanthrene to sorption sites of different energies and sequestration inside the micropores of the sediments. PMID- 19545020 TI - [Dynamics of labeled substrate N by microorganism and soil clay immobilized and its residue fractions in typical paddy soils in Dongting Lake floodplain]. AB - Dynamics of immobilization of the labeled substrate N by microorganism and soil clay and its residue fractions in soils from the plough layers of two subtropical paddy soils (Reddish clayey soil, Purple alluvial soil) in the Dongting Lake floodplain were studied. A laboratory-flooded incubation experiment was composed of three treatments: control (CK), labeled ammonium sulphate group (15NA) and the combined of labeled ammonium sulphate and rice straw group (S + 15NA). During the incubation, microbial biomass N (BN) increased firstly and then those decreased and tended to stable subsequently, while the content of fixed ammonium changed little. Native BN acted as the major N pool because the percentage-of labeled substrate BN in reddish clayey soil and purple alluvial soil were 0.30%-6.67% and 1.00%-3.47%, respectively. The combined application of rice straw and chemical fertilizer (S + 15NA) improved the immobilization of inorganic N by microorganism, because the immobilization ratio of substrate N in reddish soil and purple alluvial soil were 6.78% and 10.78%, respectively, for S + 15NA treatment, higher than those for 15NA treatment. The immobilization ratio of substrate N by soil clay in reddish clayey soil and purple alluvial soil were 2.48%-10.57% and 12.55%-30.04%, respectively. However, the immobilization ratio of substrate N by soil clay in the two soils were 7.14% and 21.53%, respectively, for S + 15NA treatment, lower than those for NA treatment. The incorporation of chemical fertilizer and rice straw increased the N remain percent. The main residue formation of the labeled substrate N was total hydrolysable N (> 72%) in Reddish clayey soil, while it was total hydrolysable N (44.0%-53.2%) and fixed ammonium (35.2%-37.5%) in Purple alluvial soil. The residue of mineral nitrogen ranged 10%-20% in the two soils. In conclusion, fertilization method and the type of soil clay had important effects on the immobilization and mineralization of substrate N. The combined application of chemical fertilizer and straws increased the immobilization of inorganic nitrogen by microorganism and decreased immobilization of inorganic nitrogen by soil clay. The combined application of chemical fertilizer and straws decreased the loss of chemical fertilizer N, increased residue of nonhydrolysable N, and decreased residue of mineral nitrogen. PMID- 19545021 TI - [Responses of soil organic carbon content and fractions to land-use conversion from paddy field to upland]. AB - Natural 13C abundance determination method coupled with physical fractionation of soil organic carbon (SOC) was used to evaluate the responses of SOC and its fractions to long-term land-use conversion from paddy field to upland field (corn cultivation). Results showed that land-use conversion from paddy field to upland field led to significant decreases in the contents of SOC and total nitrogen (TN). Concentrations of total organic carbon (TOC) and TN were respectively greater by 76.7% and 47.6% in the paddy field than those in the corn field. Concentrations of occluded particulate organic matter (oPOM) and mineral associated organic matter (MOM) on a whole soil basis were two times higher in the paddy field than those in the upland field, while no significant difference was found in free particulate organic matter (fPOM). Carbon concentrations of oPOM and MOM fractions on their own weight basis were significantly greater in the paddy field than those in the upland field, especially the oPOM fraction, which was 6 times higher in the former than that in the latter. It could be concluded that SOC protection exerted by soil aggregates in paddy soil was greater than that in upland soil. After a 19-year conversion from paddy field to corn field, delta13C values of SOC fractions significantly increased. Maize derived carbon (C) accounted for 54.6%, 24.7%, and 19.0% in fPOM, oPOM and MOM, respectively. Mean residence time (MRT) of the initial rice-derived C increased in the order fPOM (24 a) < oPOM (67 a) < MOM (90 a). The above results further indicate that paddy field soil owns greater capability of carbon sequestration than upland soil mainly through increasing the contents of oPOM and MOM in the fractions of SOC. PMID- 19545022 TI - [Temporal and spatial variations of soil respiration in an Artemisia ordosica shrubland ecosystem in Kubuqi Desert]. AB - Based on the dynamic measurements of soil respiration using a closed dynamic chamber and its related environmental factors in a desert shrubland ecosystem regularly during the growing season (May-September) of 2006, we studied the diurnal and seasonal variations of soil respiration of two different land cover soils and their responses to soil temperature, soil water content and biotic factors. The objective was to evaluate the temporal and spatial patterns of soil respiration and their responsible factors in Artemisia ordosica shrubland in Kubuqi Desert, Inner Mongolia, China. The diurnal variation of soil respiration showed an asymmetric single-peak pattern, with the peak value occurring around 12:00. Soil respiration fluctuated greatly during the growing season, reaching peak values in July-August. There was a significant linear relationship between soil respiration rate and soil water content at 10 cm depth. Most of the seasonal variation in soil respiration (75%-77%) could be explained by the variation in soil water content. The mean soil respiration under the shrub canopy was (155.58 +/- 15.20) mg x (m2 x h)(-1), which was significantly higher than that for the bare ground between the shrubs (110.50 +/- 6.77) mg x (m2 x h)(-1). The sensitivity of soil respiration to soil water content was also significantly higher for the soils under the canopy than for the bare ground soils. The spatial variation of soil respiration was caused mainly by the root biomass, which can be explained about 43% of heterogeneity. The results suggest that variation on a small time and space scales must be taken into consideration when estimating soil CO2 efflux in the desert ecosystems. PMID- 19545023 TI - [Spatial distribution and ecological significance of heavy metals in soils from Chatian mercury mining deposit, western Hunan province]. AB - Ores, waste tailings and slag, together with three typical soil profiles (natural soil profiles far from mine entrance and near mine entrance, soil profile under slag) in Chatian mercury mining deposit (CMD), western Hunan province were sampled and their concentrations of mercury (Hg), arsenic (As), lead (Pb), cadmium (Cd), zinc (Zn) were determined by HG-ICP-AES and ICP-MS. Enrichment factor and correlation analysis were taken to investigate the origins, distribution and migration of Hg, as well as other heavy metals in the CMD. The results show that Hg is enriched in the bottom of the soil profile far from mine entrance but accumulated in the surface of soil profiles near mine entrance and under slag. The soil profiles near mine entrance and under slag are both contaminated by Hg, while the latter is contaminated more heavily. In the soil profile under slag, Hg concentration in the surface soil, Hg average concentration in the total profile, and the leaching depth of soil Hg are 640 microg x g(-1), (76.74 +/- 171.71) microg x g(-1), and more than 100 cm, respectively; while 6.5 microg x g(-1), (2.74 +/- 1.90) microg x g(-1), and 40 cm, respectively, are found in the soil profile near mine entrance. Soil in the mercury mine area is also polluted by Cd, As, Pb, Zn besides metallogenic element Hg, among which Cd pollution is relatively heavier than others. The mobility of the studied heavy metals in soil follows the order as Hg > Cd > As > Zn approximately equal to Pb. The leaching depth of the heavy metals is influenced by total concentration in the surface soil and soil physico-chemical parameters. The origins, distribution and migration of heavy metals in soil profile in the mining area are related to primary geological environment, and strongly influenced by human mining activities. PMID- 19545024 TI - [Characteristics and sources of polycyclic aromatic hydrocarbons in surface soil from chemical industrial areas]. AB - Concentrations and composition of 16 polycyclic aromatic hydrocarbons (PAHs) listed for prior control by US EPA were detected by analyzing 28 surface soil samples from the chemical industrial areas of Tianjin Binhai New Area with a gas chromatography equipped with mass spectrometry (GC/MS). Relative concentrations of PAH compounds with different benzene rings and principal component analysis were used to identify the possible sources of soil PAHs. The maximum PAH concentration in all the samples was 5,991.7 ng x g(-1), with a mean values of 1,185.0 ng x g(-1). The concentrations of four and five rings PAH components were higher than that of the other PAH components in Tanggu and Hangu chemical industrial areas, and three rings PAH components were major PAHs in Dagang petroleum industrial area. There is significant positive correlation (n = 28, R2 = 0.847, p < 0.01) between PAH concentrations and total organic carbon (TOC) contents in the soil samples. Coal combustion was the dominant source of PAHs in chemical industrial areas, while petroleum volatilization and leakage were main contributors of PAHs in Dagang petroleum industrial area. PMID- 19545025 TI - [Distribution of HCB discharged from a chemical plant in plants]. AB - The distribution characteristics of hexachlorobenzene (HCB) in plant and rhizosphere soil in contamination conduit, a nearby river and a cropland were studied and the impact factors were also discussed. The results are summarized as follows: the range of the HCB concentration in plant and rhizosphere soil in investigation area were respectively from 4.45 microg x kg(-1) to 1,189.89 microg x kg(-1) (dw) and from 27.93 microg x kg(-1) to 3,480.71 microg x kg(-1) (dw). Higher enrichment of HCB in woodplant than herbs due to higher fat concentration in woodplant in the contamination conduit and the rich concentrtion factor of woodplant and herbs were 0.41-2.55 and 0.01-1.34. The range of HCB concentrations in plants in nearby croplands was significantly wide (4.45-333.1 microg x kg(-1)) while HCB concentrations in different parts of plant were various, e.g. HCB concentrations in fruit, root and shoot of taro were 318.77 microg x kg(-1), 281.02 microg x kg(-1) and 10.94 microg x kg(-1). There was a remarkable positive relation between the concentrations of HCB in plant and fat concentration of plant while no relativity between the concentrations of HCB in plant and those in ground soils in the contamination conduit and cropland. The concentration levels of HCB in plant and rhizosphere soil in river were dramatically decreased with increasing distance from contaminated conduit. There was a remarkable positive relation between the concentrations of HCB in plant and those in ground soils but no relation between concentrations of HCB in plant and fat concentration of plant in river. The distribution characteristics of HCB in plants were influenced by contaminated levels, fat concentration and Partition-transfer model. PMID- 19545026 TI - [Effects of lower concentrations of Cd on micronutrients uptake and production of phytochelatins (PCs) in Ceratophyllum demersum]. AB - A solution experiment with a range of lower cadmium concentrations (0.01-0.64 micromol/L) was conducted to study micronutrients uptake and production of phytochelatins (PCs) in a submerged aquatic plant Ceratophyllum demersum after exposure for 7 d, 14 d, 21 d. The results showed the uptake of Cu, Zn in Ceratophyllum demersum was promoted when exposed to lower concentrations of cadmium, whereas Mn uptake was inhibited. Unlike Cu, Zn and Mn, the content of Fe was unaffected by Cd uptake. Our data showed that Ceratophyllum demersum had stronger Cd tolerance and obvious absorption and cumulative effects to external lower concentrations of Cd exposure. The inhibition of growth was observed after 21 d in 0.08 micromol/L Cd. The production of PCs was significantly induced by 0.02-0.64 micromol/L Cd for 7 d (p < 0.05), while declined after exposure for 14 d, 21 d. The good dose-related response of PCs was observed in manner of linear for 7 d and parabola for 14 d and 21 d. Regressive analysis showed that there was a significantly positive correlation between PC contents and Cd toxicity in Ceratophyllum demersum. The results suggested that PCs could be a sensitive biomarker for estimating Cd phytotoxicity and subsequently a qualitative tool for studies on Cd contamination. PMID- 19545027 TI - [Microbial community constitute and phylogenetic analysis on oil polluted soils in oilfields of China]. AB - The microbial communities in oil polluted soils from oilfields of China were analyzed using a modern molecular biological approach. The crude total DNA from soils were directly extracted, 16S rDNA fraction were amplified by PCR, the bands were separated using denaturing gradient gel electrophoresis (DGGE), and some of the typical bands were sequenced and compared with the records of NCBI to classify the germs. Comparison of the DNA extraction indicated that the yields were 1.4-2.2 times as much as the existing technique, and the DNA purity reached 1.8-2.0. With higher similarity and correlation in CQ and DQ oilfields but lower in that of SL and YM oilfields, the influence factors to microbial communities consisted in the elementary properties of soil samples such as oil contents, water ratio, etc. Extraction and sequencing analysis of selected 16S rDNA bands demonstrated a range of similarity of 89%-100% to reference bacteria. The Shannon Weiner index of the oil contaminated soils was 0.5-1.2, and which was increased slightly along with the enhancement of microbial amounts and FDA activities. According to this research, it has possibility in developing a reliable tool for researching oil contaminated soil bacteria diversity, and in applying a credible foundation for adjusting the microbial ecosystem and determining the dominant community in contaminated soils in oilfields. PMID- 19545028 TI - [Isolation and identification of a 4-aminobenzenesulphonate biodegrading strain and its degradation characteristics]. AB - A 4-aminobenzenesulphonate-degrading strain W1 was isolated from the activated sludge of Dalian municipal wastewater treatment plant. Strain W1 was able to utilize 4-ABS as sole carbon and energy source under aerobic condition. It was identified as Pannonibacter sp. according to its morphological, physiological and biochemical characteristics and the analysis of its 16S rDNA gene. The optimum conditions for 4-ABS biodegradation in the shaking flasks were 10% inoculum, 30 degrees C, pH 7.0 and were rotated at speed 150 r/min, respectively. Strain W1 could keep high degrading ability even in the presence of extra carbon source. Release of ammonia and sulfate were around 77.6% and 91.5% of theoretic concentration according to 4-ABS degradation, for 4-ABS could serve as the nitrogen and sulfur source. Strain W1 showed efficient biodegradability even at 2,500 mg/L, and 90% 4-ABS removal was achieved in 32 h. The 84.4% TOC removal was achieved after 94.7% consumption of 4-ABS, and no aromatic intermediates were detected, referring the complete mineralization of 4-ABS by strain W1. PMID- 19545029 TI - [Degradation of dichlorvos by Rhodobacter sphaeroides]. AB - Rhodobacter sphaeroides possesses an extensive range of energy acquiring mechanisms including photosynthesis, lithotrophy, aerobic and anaerobic respiration. It can produce 5-aminolevulinic acid, CoQ10, carotenoids, hydrogen, etc. by fermentation. A Rhodobacter sphaeroides strain was isolated, designated as EBL0706, from soil. The degradation of dichlorvos (DDVP) by the Rhodobacter sphaeroides was investigated. 98% of DDVP could be degraded in water solution in 12 h when 5 x 10(8) CFU/mL Rhodobacter sphaeroides was added to 400 mg/L DDVP solution under pH 6.9-7.5 and 20-50 degrees C. This strain could also degrade the DDVP residues on Chinese cabbage leaves effectively. PMID- 19545030 TI - [Interaction between calcium and lead affects the toxicity to embryo of zebrafish (Danio rerio)]. AB - This study tested the hypothesis that increased Ca2+ content increases the sensitivity of the developing embryos and larvae of zebrafish (Danio rerio) to Pb. And the aim of the study was to investigate the extent to which calcium can individually mitigate lead ion toxicity based on the concept of biotic ligand model (BLM). Embryos of the zebrafish were exposed to various Pb concentrations. Chemical characteristics of water and representative toxicological endpoints of zebrafish embryo were recorded. And general growth retardation as a major toxicological endpoint was used for analysis at 72 h due to its sensitivity and facility. The BLM software of Visual MINTEQ (Version 2.5.2) was employed to calculate the chemical speciation in the solution. The results showed that when Ca2+ concentration increased from 0.25 mmol/L to 2.00 mmol/L, the toxicity of lead on embryos of zebrafish (Danio rerio) decreased markedly after 72 h. And a large part of these decrease can be explained by the positive linear relations between EC50{Pb2+}/EC50[Pb]T (expressed as lead ion activity/dissolved total concentration) and activity/total concentration of Ca2+, through which the influence of Ca2+ on toxicity could be predicted. The results support the assumptions of the BLM and associated with competition between lead and calcium for binding on transport and toxic action sites on biological surfaces. However, when Ca2+ concentration increased from 2.00 mmol/L to 4.00 mmol/L, the toxicity of lead on embryos of zebrafish (Danio rerio) seemed to be constant at 72 h. PMID- 19545031 TI - [Acute toxicity analysis performance of CellSense biosensor with E. coli]. AB - E. coli microbial electrodes for CellSense biosensor were prepared by polycarbonate membrane immobilization process, and their performance for heavy metals and toxic organic compounds acute toxicity determination were studied. The results showed that when E. coli was in logarithmic and stationary phase, the CellSense biosensor with E. coli showed good performance in heavy metal ions and organic pollutants acute toxicity analysis, when E. coli was in its decline phase, the stability and sensitivity of the CellSense biosensor was poor. The EC50 values of Hg2+, Cu2+, Zn2+, o-chlorophenol (2-CP) and p-nitrophenol (4-NP) detected by CellSense biosensor with E. coli were 0.6, 3.1, 5.8, 180 and 94 microg/mL, respectively. The immobilized E. coli electrodes could still suit for acute toxicity assessment after 2 months storage at 4 degrees C. PMID- 19545032 TI - [Mechanisms of bioelectricity generation in Enterobacter aerogenes-based microbial fuel cells]. AB - Microbial fuel cells (MFCs) using hydrogen-producing bacteria (HPB) could utilize a large number of substrates to generate power. However, the coulombic efficiency is limited by the fact that only suspended cells are used as biocatalyst in anodic medium. MFCs using Fe (III)-reducing bacteria have high energy recovery efficiency, but can only utilize some simple organic matters. In this study, Enterobacter aerogenes XM02, a hydrogen-producing strain with Fe(III)-reducing activity, was selected as biocatalyst for MFCs, which could produce electricity by digesting lots of carbohydrates even starch. Graphite felt, a material with high specific surface area and hydrogen catalysis, instead of carbon paper supported platinum, was used as anode material. The coulombic efficiency had been substantially improved from 1.68% to 42.49%, higher than other HPB-based MFCs previously reported. The SEM image proved the ability of XM02 strain to colonize on the anode surface. Power generation of MFCs could restore quickly when anodic medium was completely replaced with non-growth medium containing glucose. This suggested that the attached cells contributed to electricity production because planktonic cells had been removed during the medium replacement. This study proposed the mechanism of power generated from in situ oxidation of hydrogen produced by the XM02 strain biofilm. PMID- 19545033 TI - [Electricity generation using high concentration terephthalic acid solution by microbial fuel cell]. AB - The high concentration terephthalic acid (TA) solution as the substrate of microbial fuel cell (MFC) was studied to generate electricity. The open circuit voltage was 0.54 V after inoculating for 210 h with anaerobic activated sludge, which proved that TA can be the substrate of microbial fuel cell to generate electricity. The influence of pH and substrate concentration on generating electricity was studied deeply. The voltage output of external resistance (R = 1,000 Omega) was the highest when pH was 8.0. It increased as the substrate concentration increasing and tended towards a maximum value. The maximum voltage output Umax was 0.5 V and Ks was 785.2 mg/L by Monod equation regression. When the substrate concentration (according to COD) was 4000 mg/L, the maximum power density was 96.3 mW/m2, coulomb efficiency was 2.66% and COD removal rate was 80.3%. PMID- 19545034 TI - [Electricity production from surplus sludge using microbial fuel cells]. AB - A single-chamber and membrane-less microbial fuel cells were successfully started up using anaerobic sludge as inoculums without any chemical substance for 20 d. The electricity generation of the microbial fuel cell using surplus sludge as fuel and the change of substrate were investigated. The results showed that the obtained maximum voltage and power density were 495 mV and 44 mW x m(-2) (fixed 1,000 Omega), and the internal resistance was about 300 Omega during steady state. In a cycle, the removal efficiency of SS and VSS were 27.3% and 28.7%, pH was 6.5-8.0. In addition, the COD increased from 617 mg x L(-1) to 1,150 mg x L( 1) and decreased afterwards with time. The change of glucose was similar to that of COD, glucose increased from 47 mg x L(-1) to 60 mg x L(-1) and decreased afterwards with time. Consequently, the microbial fuel cell can transform chemical energy of surplus sludge into the cleanest electrical energy, and it provides a new way of sludge recycling. PMID- 19545035 TI - [Effect of additive and washing pre-treatment on heavy metal vaporization during sintering process of MSWI fly ash]. AB - An experimental work was carried out to investigate the effect of additives (clay and coal fly ash) and washing-pretreatment on the stabilization of critical heavy metals (Cd, Cu, Zn, Pb, Cr, Ni) during a sintering process of municipal solid waste incinerator (MSWI) fly ash. The proportions of the three constituents were varied to adjust the mixture compositions. The washing time were 8 h, 16 h and 24 h. The material was compacted in cylindrical specimens at 3 kN and treated at 1,100 degrees C for 4 h. When the clay content was increased to 70%, the stabilized ratio was increased from 16.96% to 28.42% for Cd, from 10.58% to 37.02% for Pb, from 46.38% to 55.14% for Cu, from 42.14% to 64.47% for Zn, but the stabilized ratio of Ni and Cr was decreased. When coal fly ash was increased in the MSWI fly ash, the stabilized ratio was decreased from 16.96% to 4.67% for Cd, increased from 46.86% to 81.43% for Cu, but the addition of coal fly ash did not increased the stabilized ratio of Zn, Pb, Ni and Cr. Washing pre-treatment increased the stabilized ratio of Cd, Cu and Pb significantly. The leaching behavior of the heavy metals in the sintered products was studied by the toxicity characteristic leaching procedure (TCLP), the results showed that the leaching concentration of the six heavy metals were very low. PMID- 19545036 TI - [Impacts of different aeration modes on nutrients conservation during swine manure composting with magnesium salt addition]. AB - Experiments were carried out to investigate the effect of two aeration modes on nutrients conservation in the thermophilic stage of swine manure composting with adding magnesium chloride. These results indicated that in the end of thermophilic stage of composting, the ammonia nitrogen losses of the piles with the intermittent aeration and with continuous aeration, were 23.56 g and 56.98 g, respectively, which means the loss of ammonia nitrogen of swine composting by the intermittent aeration was just 41.35% of that by the continuous aeration. Such loss of ammonia nitrogen resulted in 9.8% higher of the total kjeldahl nitrogen (TKN) concentration in the pile with the intermittent aeration than that with the continuous aeration. No significant difference occurred between two piles in orthophosphate, sequential extraction phosphorus and the total phosphorus (TP). However, in the pile with intermittent aeration, the percentage of the easily dissolved phosphorous forms such as H2O-P and NaHCO3-P was increased from 27.6% of TP to 66.5%, and the other pile from 27.6% to 64.9%. The TP concentrations in both piles were 17.2 g/kg in the end of thermophilic composting stage. The mixed crystals containing magnesium and phosphorus were formed in both piles of swine composting. PMID- 19545037 TI - [Theoretical thinking in sustainable city construction of China]. AB - Based on the concept of ecosystem services and welfare, a definition of a sustainable city is proposed, and a quantitative model to describe sustainable welfare is established. With the analysis of the major issues and driving forces of sustainable city construction in China, the approaches for sustainable city construction are proposed as follows: to promote study on the theory and methodology of sustainable city construction, to set up and perfect institutional and management systems, and to establish a performance assessment system and an effective operation mechanism for sustainable city construction in China. PMID- 19545038 TI - [Evaluation of malaria and vector-control measures]. PMID- 19545039 TI - [Evaluation of the anopheline mosquito aggressivity and of malaria transmission risk: methods used in French Army]. AB - Identification of the anopheline mosquito species involved in local transmission as well as knowledge of its biology and behavior is necessary for malaria vector control. To allow such study, two methods are usually used to capture adult mosquitoes, i.e., night catches on human volunteers and light-trap collections with human bait. The purpose of this article is to describe these two methods including their advantages and disadvantages as well as a method of surveying breeding sites as implemented by French Army personnel. PMID- 19545040 TI - [Concept, application and use of verbal autopsy]. AB - Verbal autopsy (VA) is a technique designed for reliable determination of the cause of death by interviewing the entourage of the deceased. When conducted rigorously using a standardized questionnaire, VA can establish the most probable diagnosis. Data obtained by VA can be suitable for demographic or epidemiologic purposes. The VA technique depends on the exact purpose for which the data will be used. PMID- 19545041 TI - [Contribution of remote sensing to malaria control]. AB - Despite national and international efforts, malaria remains a major public health problem and the fight to control the disease is confronted by numerous hurdles. Study of space and time dynamics of malaria is necessary as a basis for making appropriate decision and prioritizing intervention including in areas where field data are rare and sanitary information systems are inadequate. Evaluation of malarial risk should also help anticipate the risk of epidemics as a basis for early warning systems. Since 1960-70 civilian satellites launched for earth observation have been providing information for the measuring or evaluating geo climatic and anthropogenic factors related to malaria transmission and burden. Remotely sensed data gathered for several civilian or military studies have allowed setup of entomological, parasitological, and epidemiological risk models and maps for rural and urban areas. Mapping of human populations at risk has also benefited from remotely sensing. The results of the published studies show that remote sensing is a suitable tool for optimizing planning, efficacy and efficiency of malaria control. PMID- 19545043 TI - [Vector control for armed forces: a historical requirement requiring continual adaptation]. AB - From time immemorial vector-borne diseases have decimated the fighting capacity of armies and caused suspension or cancellation of military operations. For this reason vector control strategies have always been a major requirement in ensuring the operational readiness of armed forces. Using malaria as an example, this article provides an update on vector control methods used by the French army in terms of available tools and of their applications in function of the operational situation. In coming years vector control strategies must be modernized not only by assessing vector-borne disease threats in the field and improving resources available to troops but also by professionalizing this service by creating specialized units. PMID- 19545042 TI - [Vector control methods against malaria and vector resistance to insecticides in Africa]. AB - Vector control methods against malaria must be adapted to the targeted vectors. Control methods against malaria currently depend on deploying insecticide-treated nets, indoor spraying of remanent insecticides and, to a lesser extent, eliminating larval breeding sites. The remanence of the insecticidal effect and efficacy of the vector control must be evaluated. Vector resistance to insecticides is one of the main limitations for use. PMID- 19545044 TI - [Evaluation of vector control techniques: an economic approach]. AB - Programs of disease control must be evaluated. Evaluation of these programs in terms of public health is straightforward but should be carried out in conjunction with economic evaluation to measure the cost effectiveness and cost benefit ratio. The purpose of this report is to clarify the importance of economic evaluation by explaining why it is necessary, when it should be carried out, and what methods should be used. Program evaluation is a process with several steps. Each step is associated with specific indicators. Determining these indicators is a prerequisite for construction of the database needed for evaluation. Two methods are proposed for data analysis, i.e., a simple one for cost-effectiveness and cost-benefit analysis and a more sophisticated one for impact analysis that must take into consideration treated and non-treated groups. Economic evaluation of disease control programs is as important as epidemiologic evaluation. However this evaluation cannot be carried out as a standalone procedure but only within a multidisciplinary framework. PMID- 19545045 TI - [Will climate and demography have a major impact on malaria in sub-Saharan Africa in the next 20 years?]. AB - The purpose of this review of the literature is to present factors possibly affecting the spread of malaria in sub-Saharan Africa over the next 20 years. Malaria is a vector-borne disease that depends on environmental and human constraints. The main environmental limitations involve susceptibility of the vector (mosquitoes of the Anopheles genus) and parasite (Plasmodium falciparum) to climate. Malaria is a stable, endemic disease over most of the African continent. Climatic change can only affect a few regions on the fringes of stable zones (e.g. altitude areas or Sahel) where malaria is an unstable, epidemic disease. Higher temperatures could induce a decrease of malaria transmission in regions of the Sahel or an increase in the highlands. The extent of these overall trends will depend on the unpredictable occurrence of major meteorological phenomenon as well as on human activities affecting the environment that could lead to dramatic but limited outbreaks in some locations. The most influential human factors could be runaway demographic growth and urban development. Estimations based on modeling studies indicate that urbanization will lead to a 53.5% drop in exposure to malaria by 2030. However this reduction could be less than expected because of adaptation of Anopheles gambiae and An. arabiensis, the main vectors of malaria in sub-Saharan Africa, to the urban environment as well as increasing vector resistance to insecticides. Another unforeseeable factor that could induce unexpected malaria epidemics is mass migration due to war or famine. Finally immunosuppressive illnesses (e.g. HIV and malnutrition) could alter individual susceptibility to malaria. Social constraints also include human activities that modify land use. In this regard land use (e.g. forest clearance and irrigation) is known to influence the burden of malaria that is itself dependent on local determinants of transmission. Overall the most important social constraint for the population will be access to malarial prevention and implementation action to control this scourge. PMID- 19545046 TI - [Hepatobiliary transport of glutathione and its role in cholestasis]. AB - Glutathione is a tripeptide comprised by L-glutamate, L-cysteine, and glycine, that serves antioxygenation and deintoxication functions within the cell. Recent study has found that glutathione is the main driving force for bile salt independent bile flow, impaired biliary excretion of glutathione can lead to cholestasis. This review focuses on hepatobiliary transport of glutathione and its role in cholestasis. Based on the evidence of choleretic effect of glutathione, enhancement of biliary excretion of glutathione may be a good strategy for prevention and treatment of cholestasis. PMID- 19545047 TI - [Progress in the study of drug delivery system based on nanoparticles to overcome multi-drug resistance]. AB - Multi-drug resistance (MDR) of cancer cells is a major cause of failure in chemotherapy. To the majority of anti-cancer drugs, tumor cells are able to generate a multi-drug resistance; but there is no common views on the mechanism of MDR. This review summarizes the use of drug delivery system based on nanoparticles to overcome MDR in recent years. Three kinds including non modified, ligand-modified and multifunctional drug delivery systems are described. Especially, the mechanism of reversing MDR based on nanoparticles is covered. Through efficiently offsetting and antagonizing the action of pumping drugs out of the tumor cells, drug delivery system based on nanoparticles can increase the concentration of the drug in tumors, while reduce the side effects on normal cells and overcome multi-drug resistance. The use of drug-loaded nanoparticles, which combines nanotechnology with the strategy of active and passive targeting administration, has shown significant prospect improving cancer therapy. PMID- 19545048 TI - [Neuroprotective effect of batroxobin on experimental intracerebral hemorrhage in rats]. AB - This study is to investigate if batroxobin has the protective effect against nerve injury caused by cerebral hemorrhage in rats and its possible mechanism. Animals were divided into sham group, model group, batroxobin 4, 8, and 16 BU x kg(-1) groups and nimodipine positive control group. On the brain stereotaxic apparatus, the rat intracerebral hemorrhage model was established by injecting collagenase with microinjector into the brain caudate nucleus which was located according to the brain stereotaxic atlas. Neuroethology of the rats was estimated. The brain tissue pathomorphology was observed with electron microscope. The water content of the brain tissue was quantitated with wet/dry weight measurement. SOD and MDA were determined according to the kit procedure, and free Ca2+ concentration in neurocyte was measured by fluorospectrophotometer. As shown in results, batroxobin could improve neuroethology scale of the rats, relieve histiocyte edema and bleeding degree. The water content of the brain tissue, MDA and free Ca2+ concentration were reduced and SOD activity was raised in batroxobin treatment groups. Therefore, it is possible that batroxobin has some protective effect against nerve injury caused by cerebral hemorrhage in rats, and its mechanism maybe relate to lessening brain edema, reducing MDA content, raising SOD activity, and inhibiting calcium overload. PMID- 19545049 TI - Protective effect of propofol against intracerebral hemorrhage injury in rats. AB - The neuroprotective effect of propofol against intracerebral hemorrhage (ICH) in rats was investigated. ICH was induced in rats by infusion of collagenase (Type VII) 0.5 U (1 U x microL(-1)) into the left caudate nucleus. Three doses of propofol were given intraperitoneally (i.p.) 10 min before collagenase infusion. Effects of propofol on neurological behavioral scores, brain water content (BWC), activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) in brain tissue, expression level of caspase-3 were studied. In propofol groups (30 and 100 mg x kg(-1)), the neurological behavioral score, BWC and the content of MDA were significantly lower than those in ICH group (P < 0.05, P < 0.01), whereas the activity of SOD was higher than that in ICH group (P < 0.05). Meanwhile, propofol (15, 30, and 100 mg x kg(-1)) inhibited caspase-3 expression in dose-dependent manner (r = 0.877). Brain damages caused by ICH in rats can be alleviated by propofol, which mechanism might be attributed to its antioxidant activity. PMID- 19545050 TI - [Phenylhexyl isothiocyanate induces gene p15 demethylation by down-regulating DNA methyltransferases in Molt-4 cells]. AB - This study is to investigate the effect of phenylhexyl isothiocyanate (PHI), which has been proved to be a novel histone deacetylase inhibitor (HDACi) recently, on gene p15 de novo expression in acute leukemia cell line Molt-4, and to further study its potential mechanism. Modified methylation specific PCR (MSP) was used to screen p15-M and p15-U mRNA. DNA methyltransferasel (DNMT1), 3A (DNMT3A), 3B (DNMT3B) and p15 mRNA were measured by RT-PCR. P15 protein was detected by Western blotting. Hypermethylation of gene p15 was reversed and activation transcription of gene p15 in Molt-4 was de novo after 5 days exposure to PHI in a concentration dependent manner. DNMT1 and DNMT3B were inhibited by exposure to PHI for 5 days (P < 0.05). Alteration of DNMT3A was not significant. It is showed that PHI could reverse hypermethylation of gene p15 and transcriptional activation of gene p15 is de novo by PHI. It may result from down regulating DNA methyltransferases, DNMT1 and DNMT3B, or up-regulating the histone acetylation that allows chromatin unfolding and the accessibility of regulators for transcriptional activation in the p15 promoter. PMID- 19545052 TI - [MPP+ decreased BDNF expression in PC12 cells]. AB - The aim of this study is to investigate the neurotoxic effect and mechanism of 1 methyl-4-phenylpyridinium (MPP+) on PC12 cells. MTT assay was used to investigate cell viability, Western blotting assay was performed to observe the protein level and phosphorylation, and dual-luciferase assay was used to study the transactivation. The experiment showed that MPP+ could decrease cell viability significantly in a dose-dependent manner and could decrease BDNF protein level, depress the phosphorylation of ERK, and attenuate the phosphorylation and transactivation of CREB, which is one of transcription factors of BDNF, but did not affect the activity of CaMK II in PC12 cells. So MPP+ might decrease BDNF protein level through MAPK/ERK signal pathway. PMID- 19545051 TI - [Establishment of pharmacological evaluation system for non-nucleoside reverse transcriptase inhibitors resistant HIV-1]. AB - Consistent non-nucleoside reverse-transcriptase inhibitors (NNRTIs) resistant HIV 1 strains occurred due to the clinical use for more than ten years of efavirenz (EFV), nevirapine (NVP), and delavirdine (DLV). In this study, we established nine cell-based pharmacological models according to most NNRTIs-resistant clinical tested strains, Resistant mutations were introduced into vector, pNL4 3.Luc.R-E-, by overlapping PCR. Then, pseudovirions were produced by co transfection of VSV-G plasmid and pNL4-3.Luc.R-E- -mut. All nine recombinant VSVG/HIV-mut pseudovirions (VSVG/HIV-wt, VSVG/HIV(-K103N), VSVG/HIV(-Y181C), VSVG/HIV(-L100I,K103N), VSVG/HIV(-Y188L), VSVG/HIV(-K103N,Y181C), VSVG/HIV( K103N,P225H), VSVG/HIV(-K103N,Y188L), VSVG/HIV(-K103N,G109A) and VSVG/HIV( K103N,V108I)) had high efficient infectivity. Furthermore, they all showed resistant characteristics to EFV and NVP with IC50 changes consisting with clinical reports, not to nucleoside reverse-transcriptase inhibitors (AZT and d4T). This series safe cell-based model, which could be carried out in BSL-2 laboratory, can be used for evaluating NNRTIs candidates. PMID- 19545053 TI - [Synthesis and antibacterial activity of 7 beta-[2-(2-substituted aminothiazol-4 yl)-(Z)-2-methoxyiminoacetylamido]-3-quaternaryammoniummethyl-cephalosporins]. AB - In order to find new cephalosporin with more and more potent antibacterial activity, nine new fourth-generation cephalosporins (N1-N9) were synthesized from ethyl 2-(2-aminothiazol-4-yl)-(Z)-2-methoxyiminoacetate (1) via acylation, substitution, hydrolysis, active esterification, condensation and salt formation. The structures of compounds (N1-N9) were confirmed by IR, MS, 1H NMR and elemental analysis. The target compounds possess different antimicrobial activities against Gram-positive and Gram-negative bacteria. The preliminary results of antibacterial activities revealed that they showed better antibacterial activities against Gram-positive bacteria than cefpirome sulfate. In particular, their activities against Staphylococcus aureus and Streptococcus albus are better. PMID- 19545054 TI - [Design, synthesis and antalgic activities of aralkyl-ketone-4-piperidol derivatives]. AB - A series of aralkyl-ketone-4-piperidol derivatives were synthesized and tested for their analgesic activities. All of the novel 30 compounds were prepared from 4-piperidone and alpha-halo-aralkyl-ketone through five steps, including Boc protection, nucleophilic addition in presence of CeCl3/NaI catalyst, deprotection, condensation and salification. Their structures were confirmed by 1H NMR and HRMS. Preliminary in vivo pharmacological trials showed that most of the synthesized compounds revealed analgesic effects. Among the tested compounds, 8, 13 and 22 exhibited potent analgesic activities in both mice writhing and mice hot plate model. The three compounds have low affinity for mu, delta, kappa receptors, which is a chance to find a better precursor of non-opioid analgesic for further optimization. PMID- 19545055 TI - [Pharmacophore model of integrin alphavbeta3 antagonists]. AB - In order to generate a pharmacophore model of integrin alphavbeta3 receptor antagonists and design lead compounds which have potent and selective activity against alphavbeta3 receptor with the help of this model. Thirty compounds (four categories) with highly inhibitory activity against the integrin alphavbeta3 receptor (IC50 < 110 nmol x L(-1)), amide, piperazine, piperidine, gamma valerolactam as the intermediate junction, separately, were selected as a training set to construct a three-dimensional pharmacophore models of integrin alphavbeta3 receptor antagonists with the Catalyst software. The best pharmacophore model of integrin alphavbeta3 receptor antagonists with RMS = 0.73, Correl = 0.90, Weight = 1.17, Config = 14.00 is found out, which consisting of four features: a neg ionizable core (NI), two aliphatic hydrophobic core (HP) and an aromatic ring center (RA). Some new and easily obtained compounds with fine ADME properties and highly potent activity against alphavbeta3 receptor were designed with the new pharmacophore models. PMID- 19545056 TI - [Synthesis, solubility, lipids-lowering and liver-protection activities of sulfonated formononetin]. AB - A water-soluble compound, sodium formononetin-3'-sulfonate with good lipid lowering and liver-protection activities was synthesized. It was synthesized by sulfonation reaction, and its structure was characterized by IR, NMR and elemental analyses. The solubility of sodium formononetin-3'-sulfonate in water and n-octanol/water partition coefficient were determined by UV spectrophotometry. The lipid-lowering and liver-protection activities of sodium formononetin-3'-sulfonate were tested by using rat's high fat model induce by feeding with high fat food. The results showed that sodium formononetin-3' sulfonate not only had favorable water, solubility but also had good lipid lowering and liver-protection activities. PMID- 19545057 TI - Isolation and characterization of xanthine oxidase inhibitory constituents of Pyrenacantha staudtii. AB - Six compounds have been isolated from the leaves of Pyrenacantha staudtii, two of which are new compounds. The new compounds have been characterized as kaempherol 3-O-beta-rhamnopyranosyl (1-->6)-beta-D-glucopyranoside (1) and 4-beta glucopyranosyl-(2-furyl)-5-methy-1,2-glucopyranoside phenylmethanone (2). The known compounds are 3-pyridinecarboxylic acid (3), beta-sitosterol (4), sitosterol 3-O-beta-glucopyranoside (5) and taraxerol (6). Their structures were determined by spectroscopic and chemical evidences. The two new compounds together with 3-pyridinecarboxylic acid showed significant in vitro xanthine oxidase inhibitory activity. To the best of our knowledge, this is the first report of these compounds from this plant. PMID- 19545059 TI - [Evaluating the quality of niuhuangjiedu tablets by the systematic quantified fingerprint method]. AB - The HPLC fingerprints of Niuhuangjiedu tablets (NHJDT) were established and assessed by systematic quantified fingerprint method. The chromatographic fingerprints (CFPs) were determined by reversed-phase HPLC, in which the CFPs containing 53 co-possessing peaks were marked by choosing baicalin peak as the referential peak. Fifteen batches of NHJDT were identified with hierarchical clustering analysis by using macro qualitative similarities and macro quantitative similarities as variables. According to the results of classification, the referential fingerprint (RFP) was synthesized from 10 batches of NHJDTs. Taking the RFP for the qualified model, the whole 15 batches of NHJDTs were evaluated by the systematic quantified fingerprint method. Among the 15 batches of NHJDT, 9 batches were completely qualified, the relative variance of leveling coefficient of 1 batch was obviously higher while the contents of other 5 batches were lower. The systematic quantified fingerprint method closely integrates the systematic fingerprints with the macro qualitative similarities and macro quantitative similarities to perform the entire qualification and quantitation analyses, which can briefly and effectively assess the quality of Chinese traditional medicine. PMID- 19545058 TI - [Enzyme kinetics of ligustilide metabolism in rat liver microsomes]. AB - To study the enzyme kinetics of ligustilide metabolism and the effects of selective CYP450 inhibitors on the metabolism of ligustilide in liver microsomes of rat, a LC-MS method was established for quantitative analysis of ligustilide in liver microsomes incubation system with nitrendipine as internal standard. The determination m/z for ligustilide was 173, and for nitrendipine, 315. An optimum incubation system was found and various selective CYP inhibitors were used to investigate their inhibitory effects on the metabolism of ligustilide. The results showed that enzyme kinetics of ligustilide could be significantly inhibited by ketoconazole, trimethoprim and a-naphthoflavon but scarcely inhibited by omeprazole, 4-methylpyrazole and quinidine. Therefore, CYP3A4, CYP2C9 and CYP1A2 are the major isoenzyme participated in in vitro metabolism of ligustilide. PMID- 19545060 TI - Comparison of different pharmacodynamic models for pharmacokinetic pharmacodynamic (PK-PD) modeling of carvedilol. AB - The paper is aimed to investigate the pharmacokinetic (PK) and the pharmacodynamic (PD) properties of carvedilol using indirect response and effect compartment link models, and compare the fitness of PK-PD models. Twenty male healthy Chinese volunteers received a single oral dose of 20 mg of carvedilol. The plasma concentrations of carvedilol were determined by reversed-phase HPLC method with fluorescence detection, and the pharmacokinetic parameters were calculated by DAS2.0. The mean arterial blood pressure was measured and the pharmacodynamics of carvedilol was characterized by tail-cuff manometry. The main pharmacokinetic parameters of carvedilol were as follows, t1/2 (4.56 +/- 2.56) h, Cmax (46.29 +/- 21.07) ng x mL(-1), AUC(0-infinity) (173.76 +/- 87.36) ng x mL( 1) x h. The estimated Kin was (0.41 +/- 0.31)% h(-1), Kout was (0.40 +/- 0.26) h( 1), the IC50 value was (24.40 +/- 21.10) ng x mL(-1) and the area under the effect curve (AUE) was (3.82 +/- 1.46)% h for the indirect response PD model. The Ke0 was (0.35 +/- 0.27) h(-1), the EC50 was (24.30 +/- 24.30) ng x mL(-1), and the AUE was (5.65 +/- 2.54)% h for the effect-compartment model. The HPLC method can be used for the pharmacokinetic study of carvedilol. The proposed effect compartment link model provided more appropriate and better-fitting PK/PD characteristics than the indirect response model in Chinese healthy volunteers according to Akaike's information criterion values. PMID- 19545061 TI - [Evaluation with compression equations of compression behavior of pellets with different intragranular pore volumes]. AB - Microcrystalline cellulose (MCC), calcium phosphate (DCP)/MCC (4:1, w/w) and lactose (Lac)/MCC (4:1) pellets with different intragranular porosity were prepared in an extrusion-spheronizator and three volume ratios of ethanol/water were used as binder agents to prepare pellets. The compression behaviors of these pellets with different intragranular pore volume were evaluated with the parameters of Kawakita model. The results showed that high pore volume of pellets made up of MCC had the best compressibility and low pore volume of pellets had a poor compactibility. However, the compressibility of different porosity of pellets made up of DCP/MCC (4:1) or Lac/MCC (4:1) was good, but they were not significantly different. The reason might be the main compression mechanism of high porosity of MCC pellets was plastic deformation and that of DCP/MCC pellets or Lac/MCC pellets was not plastic deformation but fragmentation. These results can be observed directly by the SEM photographs. According to these results, the conclusion could be drawn that high porosity MCC pellets and different porosity DCP/MCC pellets and Lac/MCC pellets can be used as cushion granules to maintain the original shape and release characteristics of drug pellets when pellets were tabletted. PMID- 19545062 TI - [The nasal mucosa permeability and toxicity of baicalin carrier systems liposomes, beta-cyclodextrin inclusion compound, and phospholipid complex]. AB - To increase drug concentration in the head through intranasal administration, we have investigated the excised animal nasal mucosa permeability and nasal toxicity of the baicalin drug carrier systems, such as baicalin liposomes, beta cyclodextrin inclusion compound, and phospholipid complex. A transport of baicalin drug carrier systems through nasal mucosa was simulated in diffusion chamber in vitro, and swine, caprine and rabbit nasal mucosa was used, the concentration of drug in the receptor was determined by HPLC. By taking the apparent permeability coefficients as evaluation standard, investigated the isolated animal nasal mucosa permeability of different baicalin drug systems was investigated for screening the best baicalin drug carrier system through nasal cavity administration. Toxicity of baicalin and its phospholipids complex on toad palate mucosal cilia movement and rats nasal mucosa long-term toxicity were studied in vivo. The apparent permeability coefficient of three kinds of baicalin drug carrier systems was better than that of baicalin (P < 0.05), and its lag time was obviously shortened. At the same time, the apparent permeability coefficient of phospholipid complex was higher than those of other two drug carrier systems (P < 0.05). The results showed that the baicalin phospholipids complex nasal mucosa permeability was obviously superior to the other two drug systems. Baicalin phospholipids complex had no toxicity to ciliary movement, and had no irritation to rat nasal mucosa. The results show that baicalin phospholipid complex was the best baicalin drug carrier system, it could significantly enhance the permeability of baicalin across nasal mucosa, had no toxicity to nasal mucosa, and could be used for intranasal administration. PMID- 19545064 TI - [Electrophoresis and fluorospectrophotometry methods to determine the content and entrapment efficiency of siRNA in cationic liposomes]. AB - To develop different methods for determining siRNA content and the entrapment efficiency of siRNA loaded liposomes, SYBR Gold electrophoresis method and Ribogreen fluorospectrophotometry method were used respectively. SYBR Gold electrophoresis method has a good linear relation in a range at 0.2-2.0 micromol x L(-1) (R = 0.9930), and the recovery at the high, middle and low concentrations were 96.35%, 96.92%, and 100.74%, respectively (n = 3). The intra-day and inter day RSD were far below 5% (n = 5). Ribogreen fluorospectrophotometry method has a good linear relation in a range at 10-50 nmol x L(-1) (R = 0.9971), and the recovery at the high, middle and low concentrations were 98.22%, 99.88% and 99.64%, respectively (n = 3). The intra-day and inter-day RSD were far below 5% (n = 5). The content and the entrapment efficiency of three batches of siRNA cationic liposomes were 98.52%, 97.85% and 99.20%, 96.45%, respectively, with these two methods. And there is no significant difference by ANOVA. Both of the two methods are accurate, sensitive, convenient method for determination of the siRNA content and the entrapment efficiency of siRNA loaded cationic liposomes. PMID- 19545063 TI - [In situ absorption of self-microemulsifying soft capsule of volatile oil from rhizome of ligusticum chuanxiong in rats' intestine]. AB - In order to investigate the rationality of formulation, the absorption behavior of volatile oil from rhizome of ligusticum chuanxiong (VOC) was compared with that of beta-cyclodextrin inclusion complex and self-microemulsifying soft capsule (SMESC). To study the properties of intestinal absorption in situ of SMESC, a series of studies were carried out including the absorption at different concentrations, at different intestinal regions and under different bile secretion conditions. The samples of the perfusion solution were collected at certain intervals. Ligustilide (LD) was chosen as marker component of VOC and the concentrations of which in the perfusion samples were determined by HPLC method. The results demonstrated that the absorption of LD in SMESC was the best and the absorption of VOC increased apparently (P < 0.001). The absorption of LD at concentration of 400 microg x mL(-1) was better than that at 200 microg x mL(-1) and 100 microg x mL(-1) (P < 0.001), while there was no significant difference between the absorption at concentrations of 200 microg x mL(-1) and 100 microg x mL(-1). The absorption of SMESC was not apparently influenced by bile secretion. SMESC could be absorbed in whole intestinal segments. The absorption rate constants (Ka) or apparent permeability coefficients (Papp) of SMESC showed duodenum > jejunum > colon = ileum. Ka and Papp of SMESC at duodenum were significantly higher than that at the other regions of intestine (P < 0.001). PMID- 19545065 TI - [Isolation and analysis of syringin in cell suspension cultures of Saussurea involucrata]. AB - Syringin is one of the main bioactive ingredients in Saussurea involucrata. In this study, various chromatographic techniques were employed to isolate and purify syringin in the polar extraction of cell suspension cultures of S. involucrata. The structure of syringin was characterized by the analysis of spectroscopic data. A quantitative analytical method for the content of syringin in cultures of S. involucrata was established with RP-HPLC. The method is convenient, accurate and reliable. All this results provided a basis for further studies on application of cell suspension cultures of S. involucrata for large scale production of bioactive compound syringin. PMID- 19545066 TI - [Preliminary exploration of inhibitory activities of Salvia species on PTP1B]. AB - An in vitro screening model was applied to test the inhibitory activities of 17 Salvia species on protein tyrosine phosphatase 1B (PTP1B). Root methanol extracts from wild-collected Salvia species were analyzed using this model. Most of the samples tested showed positive activities on human PTP1B. The inhibition rates of Salvia crude extracts varied from 9.76% to 100% at 30 microg x mL(-1), with the most convincing effects coming from Salvia evansiana and Salvia castanea. HPLC analysis revealed seven components shared by Salvia samples could be related to the inhibitory activities. PMID- 19545067 TI - [On the electrophysiopathological concept and ECG manifestations of ischemia, injury and necrosis]. PMID- 19545068 TI - Ten-year clinical and echocardiographic follow-up of patients undergoing percutaneous mitral commissurotomy with Inoue balloon. AB - Percutaneous mitral commissurotomy (PMC) has emerged as an effective nonsurgical technique for the treatment of patients with symptomatic mitral stenosis. This report highlights the immediate and long-term follow-up results of this procedure in an unselected cohort of patients with rheumatic mitral stenosis from a single center. PMC with Inoue balloon was performed in 70 patients in a 2-year period (1993-1994). Age of patients ranged from 18 to 67 years (mean 38 +/- 11). Atrial fibrillation was present in 18 (30%) patients. A detailed clinical and echocardiographic (two-dimensional, continuous-wave Doppler and color-flow imaging) assessment was done at follow-up. The procedure was technically successful in 61 (85%) patients with an increase in mitral valve area (MVA) from 0.96 +/- 0.2 to 1.7 +/- 0.28 cm2 (P < .001) and a reduction in mean trans-mitral gradient from 14.3 +/- 4.8 to 6.0 +/- 2.8 mmHg (P < .01). Mitral regurgitation appeared or worsened in 25 (30%) patients, of which 3 (4%) developed severe mitral regurgitation. Urgent mitral valve replacement was performed in these 3 patients. Data of 52 patients followed over a period of 105 +/- 10 months revealed MVA of 1.4 +/- 0.4 cm. Elective mitral valve replacement was done in 14 (23%) patients. Mitral restenosis diagnosed with echocardiography was seen in 24 (50%) patients, of which 14 were having recurrence of class III or more symptoms and were treated with surgery. Thus, percutaneous mitral commissurotomy is an effective and safe procedure and over 2/3 of the patients were event-free at the end of follow-up. The benefits are sustained in most of these patients on long term follow-up. PMID- 19545069 TI - [Kawasaki disease: clinical behaviour and cardiovascular complications in children in a tertiary-care level hospital]. AB - OBJECTIVES: To describe the clinical characteristics, outcome, and treatment response in a series of patients with Kawasaki disease (KD). METHODS: Case series, review of clinical records of children with KD diagnosis cared for from november 1999 to september 2006. RESULTS: 22 patients were included, male:female ratio, 1.4:1, 82% younger than 5 years, 14% with atypical presentation. Of the 22 patients, 10 (45%) received intravenous gammaglobulin (IVGG) in the first 10 days of symptoms onset at the recommended dose; four of them had coronary artery damage but none developed residual aneurysmatic lesions. Eight received IVGG in a different dose or after 10 days, six of them had coronary lesions and in two the damage was permanent. Four patients did not receive IVGG, and two had residual aneurysmatic lesions. None of those who received adequate treatment developed coronary residual damage, in comparison with 33% who received inadequate or no treatment, but difference was not statistically significant (P =.06). CONCLUSIONS: KD is a rare clinical entity in our country; even though most of the cases occurred with classic clinical criteria, late referral and delay of treatment worsen the prognosis. PMID- 19545070 TI - [Capillary blood gas test usefulness to evaluate gas exchange with 21% and 100% of oxygen inspired fractions in subjects with stable cardiopulmonary disease at 2,240 meters above sea level]. AB - OBJECTIVE: Capillary blood gas test has had ample use in the infantile population. In the adult population, the information is limited and controversial. The agreement between capillary-arterial gases seems to parallel the pH and the carbon dioxide pressure in different studied populations. In order to know the degree of agreement between these gases, we evaluate them at breathing room air and at 100% of oxygen fractions at 2,240 meters above sea level. METHODS: We obtained capillary-arterial blood gases simultaneously from subjects with stable cardiopulmonary disease in both conditions of inspired oxygen. Demographic, hemodynamic, diagnostic, and laboratory variables were gathered. STATISTICAL ANALYSIS: agreement was analyzed with the intraclass correlation coefficient and the Bland-Altman procedure. RESULTS: We studied 101 subjects, 48 men and 53 women, whose respective ages were 55 +/- 16 and 56 +/- 16. Mean systemic arterial pressure was 94.96 +/- 10.57 mmHg. Hemoglobin was 15.94 +/- 2.48 g/dl. The agreement between the variables with the inspired oxygen fractions, 21%, 100%, and the mean difference in parenthesis was respectively: potential hydrogen, 0.94 (0.0091), 0.94 (0.0039); oxygen pressure, 0.90 (2.94), 0.84 (74.99); carbon dioxide pressure, 0.97 (0.079), 0.97 (0.179); bicarbonate, 0.93 (-0.067), 0,96 (0.262); total dissolved carbon dioxide, 0.94 (-0.142), 0.93 (0.161); base excess: 0.94, (-0.125), 0.92 (0.235); oxygen saturation, 0.98 (0.764), 0.97(0.202). CONCLUSIONS: Capillary blood gas test could be a useful alternative to the arterial one, nevertheless, it is limited by its low agreement with the oxygen pressure in both oxygen inspired fractions. PMID- 19545071 TI - [The echocardiography in cardiac remodeling after acute myocardial infarction]. AB - OBJECTIVES: The goal of this study was to assess the impact of left ventricular diastolic filling on remodeling and survival after acute myocardial infarction. METHODS: We studied 36 patients with first acute myocardial treated with thrombolytic agents. A Doppler echocardiography was performed at 24 hours, 30 and 90 days after infarction. It measured the relation between E and A waves peak velocities (E/A ratio) and of the E deceleration time (EDT, ms), as well as the flow propagation velocity using color mode M and the E'-wave by tissular Doppler at the lateral mitral ring. RESULTS: Patients were divided into three groups. Group I, restrctive filling (deceleration time < 140 ms, E/FPV > or = 2, E/A> 2 and E/E' > 15). Group II, elevated filling pressure (deceleration time > or = 140 ms, E/FVP > or = 2, E/A 1,2 and E/E' < or = 15). Group III, normal filling pressure (deceleration time > or = 140 ms, E/FVP < 2, E/A < 1 and E/E' < 15). The E/FPV showed a better correlation in the group with restrictive filling and left ventricular filling pressure was significantly greater than in the group with normal filling pressure at 90 days (2.18 +/- 0.90 vs. 1.5 +/- 0.35; r = 0.99; P = .0001). The end diastolic volume (EDV) was similar in the three groups 24 hours after infarction. EDV varied at 90 days after infarction in those patients that underwent successful coronary angioplasty. Group I, 142.48 +/- 32 vs. 112.48 +/- 32, r = 573; P < .0001). CONCLUSIONS: E/FVP, using color M-mode Doppler echocardiography, estimates left ventricular filling pressure and predicts left ventricular dilation after acute myocardial infarction. PMID- 19545072 TI - [Percutaneous transluminal septal myocardial ablation in patients with obstructive septal hypertrophic myocardiopathy. Acute results and 3-year follow up]. AB - OBJECTIVE: To report the acute and Long term results of percutaneous transluminal septal myocardial ablation (PTSMA), for the treatment of hypertrophic obstructive cardiomyopathy (HOCM). PTSMA has been considered as an alternative to surgical treatment in patients with HOCM and drug-refractory symptoms or unresponsive to the implantation of a pacemaker. Acute response is generally satisfactory, but long term results have not been sufficiently described. METHODS: We did PTSMA in eight patients, with functional class III-IV of NYHA, and with a transaortic gradient at rest > or = 40 mmHg. ALcohol was administered to seven of them, and small absorbant gelatin sponge particles in the other. Acute and long term clinical and echocardiographical follow-up was done. RESULTS: During the procedure the left ventricular outflow tract (LVOT) gradient at rest was reduced significantly and the systolic anterior motion of the mitral valve (SAM) also. Five remained on functional class I, and two in class II. One patient died one year later of sepsis unrelated to the cardiopathy. CONCLUSIONS: PTSMA is an excellent non surgical option to reduce the LOVT gradient and the consequent symptoms in this patients. Long term follow-up shows they remain in a good functional class and their evolution is free of cardiovascular complications. PMID- 19545073 TI - [Ebstein's anomaly and interventricular communication, a rare association]. AB - The Ebstein's anomaly is a malformation of the tricuspid valve, in which the septal and posterior leaflets are attached to the wall of the right ventricle. The usual association is with an atrial septal defect, followed by pulmonary stenosis and pulmonary atresia; the ventricular septal defect is unusual. We present three cases with diagnosis of Ebstein's anomaly and ventricular septal defect. The initial presentation of the first two was an acyanotic heart defect with congestive heart failure and increased pulmonary flow, whereas the third patient was cyanotic and functionally impaired. The three patients underwent surgery. In the first one, the ventricular septal defect was corrected with a valvular patch, a pleat of the atrialized portion, and a tricuspid valvuloplasty. The outcome was aberrant; the patient had multiple arrhythmia episodes and died five days after surgery. In the second case, only the ventricular septal defect was corrected, the patient remains under treatment and is in functional class II. In the third patient, a one and a half ventricular surgery with a tricuspid prosthesis was performed; the outcome was favorable, the patient is in functional class II. The association of Ebstein's anomaly and ventricular septal defect is unusual. The increase of the pulmonary flow is due to the size and location of the ventricular defect. A good clinical judgment is needed to make a correct diagnosis and timely treatment. PMID- 19545074 TI - [Penetrating foreign body in the left ventricle with undetected chronic evolution]. AB - Penetrating cardiac trauma constitutes a serious threat for life. Less than the third part of all patients who suffer it arrives at the hospital alive, and half of them die. Most of the penetrating foreign bodies in the heart are metallic, and are frequently caused by firearms and rarely by self-injury. The accidental penetration is uncommon but inadvertent penetration is extremely rare. We present the case of a patient who suffered a closed-chest trauma and did not notice the penetration of a foreign body (copper wire fragment) in the heart. It remained lodged in the left ventricle for more than 3 months. This it is the only case reported in the literature where a strange body: a) crosses the free wall of the right ventricle, the right ventricular cavity, interventricular septum, and the mitral valve apparatus; b) occupies almost all the anteroposterior diameter of the heart, and c) did not produce acute or chronic bleeding. PMID- 19545075 TI - [Dilated myocardiopathy complicated with an apical clot secondary to a myocarditis. Report of a case]. AB - Acute myocarditis is a syndrome, until the patient is in heart failure. We report a case illustrating severe systolic dysfunction most probably due to an episode of postviral myocarditis, and the importance of echocardiogram in evaluating possible complications. PMID- 19545076 TI - [Pro-inflammatory and anti-inflammatory markers in coronary artery disease and acute ischemic coronary syndrome]. AB - Inflammation plays an important role in the development of atherosclerotic lesions, affecting several stages of the atheroma's development going from the initial leukocyte recruitment to the eventual rupture of the unstable atherosclerotic plaque. The inflammatory reactions within coronary atherosclerotic plaques influence the clinical outcome of acute coronary syndromes and coronary artery disease. Recent studies suggest that inflammation markers may reflect different aspects of the atherothrombotic process in relation to the stages of acute coronary syndrome. These markers play an important role in the risk of developing coronary artery disease, and may correlate with its severity. Some cytokines, acute phase proteins, acute phase reactants proteins, and adhesion molecules released from the inflammatory cells may reflect the inflammatory process in atherosclerotic plaques. However, it remains to be determined whether these pro- and anti-inflammation markers may confer risk or protection for cardiovascular disease, or simply reflect the underlying disease process. The analysis of the markers may be useful for the development of new strategies for coronary disease prevention and treatment. Therefore, we need a well-designed evaluation of these markers before their use in the clinical practice. PMID- 19545077 TI - [National Registry of Cardiac Rehabilitation Programs in Mexico]. AB - Cardiac Rehabilitation and secondary prevention programs are a group of therapeutic maneuvers that can reduce the adverse impact of cardiovascular disease, by using the cardiovascular risk factors reduction, through secondary prevention and exercise training therapy programs. This program started in Mexico in 1944, since then, several health institutions are working on a public or private basis, mainly in an isolated way. This article presents data about fourteen cardiac rehabilitation institutions that answered the first national registry of cardiac rehabilitation programs (RENAPREC) in 2007. On this study, we observed that these centers were mainly private; nevertheless, almost all of the referred population was attended in public health institutions. The core components for an adequate cardiac rehabilitation attention were satisfied by almost all these centers. The patients used to pay, by their own, this kind of medical practice. In our country, only the 0.58% of the population, that needed to be included on a cardiac rehabilitation program, was covered. This phenomena is due, in one hand, to the reduced number of cardiac rehabilitation centers in Mexico, but on the other hand, it happens because the primary physician do not refer all the eligible patients to this kid of programs. RENAPREC can be one first attempt to consolidate all the activities around the inter-institutional cardiac rehabilitation and secondary prevention programs in our country. PMID- 19545078 TI - [Leadership is for obvious for physicians or for a female physician]. PMID- 19545079 TI - [The basic principles of leadership]. AB - This overview of leadership research provides insights into the different leadership concepts. Early research on leadership focused on personality traits and leadership behaviour as determinants of good leadership. The recognition of leadership as a complex phenomenon resulted in concepts that examined leader characteristics and behaviour in the context of situational conditions. Modern cognitive approaches concentrated on the perception of leaders by followers and the perception of followers by leaders and the cognitive biases involved. There is a tendency in leadership research to integrate the three central aspects of leadership--person, situation and cognition--into a single framework. PMID- 19545080 TI - [Medical leadership competency]. AB - With all these changes in health care systems the physicians' professional duties are about to undergo changes as well. Especially economic, administrative and legal aspects are becoming more and more important in medical care. In order to take responsibility with respect to leadership aspects a profound professionalisation is required. The Curriculum Medical Leadership edited by the German Medical Association provides an extensive example of a framework for continuing professional development (CPD) courses in medical leadership. PMID- 19545081 TI - [Leadership in the hospital]. AB - Current concepts in leadership and governance on the level of supervisory board, management and departments are often considered as insufficient to cope with the profound structural change which actually takes place in the German health care system. While vertical and horizontal disconnecting is typical of the professional bureaucracy of hospitals, transition from functional to divisional structure further increases this risk. Accordingly, medical experts are oriented towards their professional peers and patient care on the one side; on the other side the management gets isolated and looses operative and strategic control. Several studies provide evidence for the relevance of role models to serve as agents of change, which are now developed into the concept of "Clinical Governance": evidence-based medicine, guidelines, continuous quality improvement, safety culture, resource accountability and organisational learning. The present situation makes it necessary to extend this conception, which focuses on the departmental level in an organisation with divisional features, to one of "Clinical Corporate Governance". This term, which also includes supervisory structures and the management board and is relevant for the total hospital and company, respectively, is based on the corporate governance concept. Inside the hospital, the management and the heads of the departments have to agree that (1) experts really need to be integrated into the decision process, and that (2) the outcomes of the entire hospital have to be regarded as equal or superior to the aims of a single department. The public image of the hospital should be one of a strong and reliable partner in health care and health care business on a local, regional and national level. Members of the supervisory board should clearly put corporate aspects above political and other implications and pay attention to personal independence from the leaders of the medical departments. PMID- 19545082 TI - [Management of quality or quality of management]. AB - All well-known quality management (QM) models in health care also address issues of the quality of leadership, even if they do so with varying intensity. Skills in managing organisations are a basically important quality in a top manager, but skills in leading people have been revealed as being most relevant. All the manuals of the most common quality management systems specify 'To-Do' lists and tasks for the top management--EFQM and KTQ/proCum Cert have definitively linked them with quality management. Quality needs to be assessed by the degree to which management goals are achieved. However, there is no easy approach to measure leadership abilities and explain it to the public. Since this is a real challenge and very difficult to achieve it is not a central focus of the various QM manuals. PMID- 19545083 TI - [Leadership and communication]. AB - Medical leadership requires specific communication skills in order to meet professional demands. Communicative behaviour is usually highly automated and not necessarily conscious. Managerial communication competes against elaborated but not role-specific behaviour patterns, especially in critical situations. Accordingly, competent medical leadership requires the awareness of individual communication habits as well as the knowledge and ability to use conversation techniques suitable for a specific situational context. The training of leadership-related communication techniques requires the de-automation of existing skills and a problem-oriented construction and re-automation of new communication techniques. PMID- 19545084 TI - [The influence of quality management on job satisfaction and work load--exemplary study in a German hospital]. AB - Surveys among employees are getting more and more relevant in hospital settings since an increase in both (1) efficiency and (2) quality in connection with (3) enhanced patient orientation will only be achieved, if at the same time the employees' health status and satisfaction are taken into account. Thus, the objective of this study was to compare the satisfaction of employees in a single hospital enquired in 2002 with that of 2005. Particular consideration was given to their view of quality management. Is there a correlation between employees' satisfaction, their degree of information on quality management, and their assessment of quality management? In the survey of 2005 employees were more satisfied with their work and their working conditions than in the previous inquiry conducted in 2002. They felt less mental stress, despite the declining length of hospitalisation combined with a higher turnover of in-hospital cases and with lower numbers of full-time staff. The employees' satisfaction, however, differed widely among the three departments with regard to the items "involvement with decisions" and "support by the superiors". The overall assessment of quality management is positive. Specific items such as the assessment of the management's commitment to quality management were strongly influenced by the employees' degree of information on quality management, which varies between departments. In the department with the lowest work satisfaction quality management was attributed a high potential for change and improvement. After quality management will have been implemented throughout the hospital, a new survey should be undertaken to evaluate whether quality management affects the employees' satisfaction with their work. PMID- 19545085 TI - [A systemic risk analysis of hospital management processes by medical employees- an effective basis for improving patient safety]. AB - Due to the knowledge gap that exists between patients and health care staff the quality of medical treatment usually cannot be assessed securely by patients. For an optimization of safety in treatment-related processes of medical care, the medical staff needs to be actively involved in preventive and proactive quality management. Using voluntary, confidential and non-punitive systematic employee surveys, vulnerable topics and areas in patient care revealing preventable risks can be identified at an early stage. Preventive measures to continuously optimize treatment quality can be defined by creating a risk portfolio and a priority list of vulnerable topics. Whereas critical incident reporting systems are suitable for continuous risk assessment by detecting safety-relevant single events, employee surveys permit to conduct a systematic risk analysis of all treatment related processes of patient care at any given point in time. PMID- 19545086 TI - Addressing health care market reform through an insurance exchange: essential policy components, the public plan option, and other issues to consider. AB - HEALTH INSURANCE EXCHANGE: This Issue Brief examines issues related to managed competition and the use of a health insurance exchange for the purpose of addressing cost, quality, and access to health care services. It discusses issues that must be addressed when designing an exchange in order to reform the health insurance market and also examines state efforts at health reform that use an exchange. RISK VS. PRICE COMPETITION: The basic component of managed competition is the creation an organized marketplace that brings together health insurers and consumers (either as individuals or through their employers). The sponsor of the exchange would set "rules of engagement" for participating insurers and offer consumers a menu of choices among different plans. Ultimately, the goal of a health insurance exchange is to shift the market from competition based on risk to competition based on price and quality. ADVERSE SELECTION AND AFFORDABILITY: Among the issues that need to be addressed if an exchange that uses managed competition has a realistic chance of reducing costs, improving quality, and expanding coverage: Everyone needs to be in the risk pool, with individuals required to purchase insurance or face significant financial consequences; effective risk adjustment is essential to eliminate risk selection as an insurance business model--forcing competition on costs and quality; the insurance benefit must be specific and clear--without standards governing cost sharing, covered services, and network coverage there is no way to assess whether a requirement to purchase or issue coverage has been met; and subsidies would be necessary for low-income individuals to purchase insurance. THE PUBLIC PLAN OPTION: The public plan option is shaping up to be one of the most contentious issues in the health reform debate. Proponents also believe of a public plan is necessary to drive private insurers toward true competition. Opponents view it as a step toward government-run health care and are wary of cost shifting from the public plan to private insurers. FUTURE OF EMPLOYMENT-BASED COVERAGE: The availability of a health insurance exchange may have implications for the future of the employment-based health benefits system and raises major questions for workers. Will employers provide a fixed contribution for the purchase of insurance through an exchange? Would that be large enough to purchase coverage? Would it be flat or vary by such factors as worker health status, age, and/or marital status or the presence of children? Would it be taxed? For both employers and workers, the implications are enormous. PMID- 19545087 TI - A retrospective and prospective view of orofacial myology. AB - Orofacial myofunctional disorders include specific conditions or behaviors that can have a negative impact on oral postures and functions. Historically, interest has focused on behaviors in the horizontal plane, highlighted by tongue thrusting. Currently, the scope of practice also includes tongue forward posturing, lip incompetence, open mouth rest posture, thumb and finger sucking, bruxism, and biting habits involving lips, fingers, tongue and cheeks. The common denominator for myofunctional conditions is a change in the inter-dental arch vertical rest posture dimension, the dental freeway space. The purposes of myofunctional therapy include normalizing the freeway space dimension by eliminating noxious habits or postures related to freeway space change. Improving cosmesis with a lips-together rest posture is also an important treatment goal. The clinical significance of the freeway space is explained in terms of the dental consequences of differential eruption patterns that can develop from postural modification of the freeway space. When the freeway space is opened for extended periods beyond the normal range, the tongue can act as a functional appliance and contribute to the development of anterior open bite or a Class II malocclusion. A clinical procedure is proposed for evaluating the freeway space dimension and incorporating the information into treatment planning and evaluation of treatment success. While dentistry/orthodontics has a primary focus on dental occlusion, or teeth-together relationships, orofacial myologists focus on teeth-apart behaviors and postures that can lead to, or have already resulted in malocclusion. PMID- 19545088 TI - Instrumentation and measurement procedures in orofacial myology. AB - The article describes measurements and measuring devices as they apply to assessment, therapy and research in orofacial myology and myofunctional therapy. Although the list is by no means complete, a wide range of items, their objectivity, complexity, applicability and accuracy is presented. PMID- 19545089 TI - Tongue lip and jaw differentiation and its relationship to orofacial myofunctional treatment. AB - A number of developmental changes occur in eating patterns from infancy through childhood. Initially a primitive reflexive process, deglutition develops into a complex, integrated voluntary/reflexive process. The movements of the tongue, lips and mandible are easily observed to undergo a transformation from synergistic, undifferentiated movements in the infant, to differentiated and refined movements required for biting, chewing, bolus formation and propulsion in the toddler and young child. This transformation is also crucial for the development of higher levels of articulatory precision and coordination required for verbal communication. This developmental process does not always occur in individuals exhibiting orofacial myofunctional disorders. This article will review current research in this area as well as describe how to evaluate for normal tongue, lip and jaw differentiation, and present exercises to develop these skills, which are necessary for successful outcomes in orofacial myofunctional treatment. PMID- 19545090 TI - Suffer the little children: fixed intraoral habit appliances for treating childhood thumbsucking habits: a critical review of the literature. AB - A critical review of the literature is presented covering the treatment of childhood thumbsucking habits using fixed intraoral habit appliances (hayrake, palatal crib). The habit appliances are classified into type and function. Data is tabulated for key references revealing the fragmented and distorted nature of the literature and its lack of consistency. A chronological approach is presented to confirm the confused and idiosyncratic character of the literature. Information is provided on the early work of Massler and Graber and the paradox of Mack, Korner and Reider. Haryett's seminal studies at the University of Alberta regarding aspects of the treatment used are critically reviewed. Reflections are presented on why Larsson's study, casting doubt on the wisdom of using habit appliances, continues to be ignored. The emergence of the Bluegrass Appliance is discussed in terms of its being a more humane appliance and the seeming reluctance of practitioners to apply it as a kinder form of appliance therapy. Information is reported on the pain and serious injuries inflicted on children by habit appliances. A comparison of the use of appliances in the USA is made with the UK, where fixed habit appliances are not popular. Concludes that fixed intraoral habit appliances are cruel and inflict pain and suffering on children out of all proportion to their necessity. Questions why these appliances continue to be used, implying that it could be a combination of financial inducement, professional insularity and the absence of concerted opposition from behavioural therapists. PMID- 19545091 TI - In defense of general surgery: rewards, threats, and challenges. PMID- 19545092 TI - Methicillin-resistant Staphylococcus aureus in a trauma population: does colonization predict infection? AB - Methicillin-resistant Staphylococcus aureus (MRSA) is increasingly responsible for infections in hospitalized patients. Patients colonized with MRSA appear to be at higher risk for subsequent MRSA infections than those who are not colonized. In this study, we determined MRSA colonization status of trauma patients at hospital admission and compared the incidence of subsequent MRSA infections between MRSA colonized and noncolonized patients. Collected data were entered into databases at a single, Level I trauma center over a 13-month period. Three hundred fifty-five adult trauma patients were screened for MRSA on admission to the trauma intensive care unit. The patients were categorized into two groups, those colonized with MRSA at admission and those who were not. Thirty six of 355 patients (10.1%) were colonized. Of the 319 patients not colonized, 21 (6.6%) developed MRSA infections. Twelve of 36 (33.3%) colonized patients developed MRSA infections (P < 0.001). No differences in types of MRSA infections were found between the two groups. Colonized patients who developed MRSA infections had higher death rates, 22.2 versus 5.0 per cent (P < 0.001). Patients colonized with MRSA on admission may be at higher risk for developing MRSA infections during hospitalization. MRSA screening protocols should be used to identify these at-risk patients. PMID- 19545093 TI - Surgical prophylaxis for inheritable malignant diseases: colorectal and gastric cancer. AB - Despite advances in modern medicine, cancer remains all too common and deadly. At its core, cancer is a disease of our DNA. As such, many cancers are passed from parents to children, making cancer one of the most commonly inherited diseases. Presently, we have no meaningful methods of "preventing" the malignant transformation that occurs as a result of an inherited gene, but investigators have identified several genetic mutations and subsequently developed risk reduction strategies that sometimes involve surgery. PMID- 19545094 TI - Perioperative management of cholelithiasis in patients presenting for laparoscopic Roux-en-Y gastric bypass: have we reached a consensus? AB - Obesity and rapid weight loss after bariatric surgery is associated with, the development of cholelithiasis and related complications. Several algorithms have been suggested in the management of the asymptomatic gallstones in patients presenting for weight loss surgery (WLS). Charts of patients presenting for laparoscopic Roux-en-Y (LRYGB) were retrospectively reviewed. Concomitant or delayed cholecystectomies were performed for symptomatic disease at the time of or after LRYGB, respectively. A total of 1376 patients underwent LRYGB and 21.0 per cent had a history of a cholecystectomy. An additional 2.7 per cent underwent cholecystectomy. The remaining 1050 "at-risk" patients were followed for a mean of 32.3 months and 4.9 per cent underwent delayed cholecystectomy for symptomatic disease. Of these patients, 88.5 per cent presented within 2 years of LRYGB. No significant morbidities were experienced by the "at-risk" cohort. Currently, there is no consensus in the treatment of asymptomatic cholelithiasis in patients presenting for WLS. A conservative regimen of reserving cholecystectomy for symptomatic disease is safe in patients undergoing LRYGB. Subsequent cholecystectomy was required in 4.9% with the majority of these patients presenting within 2 years of LRYGB. Further investigations in the form of randomized, prospective studies are necessary to clearly define the indications for cholecystectomy at the time of WLS. PMID- 19545095 TI - Endoscopic retrograde cholangiopancreatography gut perforations: when to wait! When to operate! AB - Most perforations of the gastrointestinal tract during endoscopic retrograde cholangiopancreatography (ERCP) can be managed nonoperatively. Identifying patients who require operative management is problematic. A clinical endoscopy database was queried for patients who sustained ERCP perforation over a 13-year period. Records were reviewed and analyzed with approval of the Institutional Review Board. During the study period, 12,817 patients underwent ERCP; 24 (0.2%) had an endoscopic perforation. Twelve patients had a retroperitoneal perforation during sphincterotomy and all were successfully managed nonoperatively. Nine of these were undergoing treatment for sphincter of Oddi dysfunction. Twelve patients had perforation remote from the papilla. Of these, 10 required surgical intervention. Six patients had surgically altered anatomy (three postpancreaticoduodenectomy, three post-Billroth II gastrectomy) and one had situs inversus. Six of these seven required surgical intervention. Median length of stay of all patients was 7.5 days, morbidity was 25 per cent, and one patient died 16 days after surgery. Gut perforation after ERCP requires prompt surgical evaluation. Patients with sphincterotomy-related retroperitoneal perforation can be managed safely with nonoperative therapy in most instances. Patients with remote perforation usually need surgical intervention. Altered foregut anatomy leads to injuries that usually require operative management. PMID- 19545096 TI - Intraoperative endoscopy and leaks after laparoscopic Roux-en-Y gastric bypass. AB - Postoperative leaks after laparoscopic Roux-en-Y gastric bypass (LRYGB) are a source of morbidity and mortality. Any intervention that would decrease leak rates after LRYGB would be useful. This investigation tested the hypothesis that postoperative leak rates are lower after LRYGB with the routine use of intraoperative endoscopy (EN). Consecutive patients who underwent LRYGB were included. Intraoperative leak testing with air and methylene blue through an orogastric tube (OG) was used in the first 200 patients. Intraoperative endoscopy was used after the first 200 patients. There were 400 patients in this study. Preoperative demographics did not differ between groups. The intraoperative leak rate of the EN group was double the OG group (8 vs 4%; P = not significant), although the difference was not statistically significant. The OG group had a postoperative leak rate of 4 per cent with a mortality rate of 1 per cent. The EN group had a postoperative leak rate of 0.5 per cent with a mortality rate of 0 per cent. The difference in leak rates was statistically significant (P < 0.04). Despite the issues of learning curve, EN demonstrates more intraoperative leaks than OG, indicating EN may be a more sensitive test than OG. Routine use of EN is associated with less postoperative leaks after LRYGB. PMID- 19545097 TI - Multimodality treatment for esophageal malignancy: the roles of surgery and neoadjuvant therapy. AB - The best curative treatment for esophageal malignancy remains controversial. In 2003, we presented our institution's experience with 124 patients treated from 1990 to 2001. Here we update that experience with an additional 6 years' data. A total of 221 patients underwent surgical resection from 1990 to 2007; 128 had up front surgery, 88 underwent surgery after neoadjuvant radiation and chemotherapy (NARCS), and five underwent surgery after neoadjuvant, single-agent therapy. Principle outcomes of interest were 30-day and in-hospital mortality as well 3- and 5-year survival rates. Overall 3- and 5-year survival rates were 38 and 33 per cent. NARCS achieved complete pathologic result in 32 per cent of patients with corresponding 3- and 5-year survival rates of 58 and 53 per cent. The 3- and 5-year survival rates for all patients undergoing NARCS were 36 and 31 per cent versus 24 and 18 per cent for patients with up-front surgery for anything over Stage I disease (P = 0.01). The 3- and 5-year survival rates for patients with up front resection of Stage I disease were 78 and 70 per cent. Overall, 30-day and in-hospital mortalities were 1.8 and 2.3 per cent. Since January 1, 2000, hospital mortality has been less than 0.8 per cent. We prefer NARCS for malignancy of the esophagus, except in those patients with high-grade dysplasia (carcinoma in situ), suspected Stage I disease, poor performance status, or urgent/emergent circumstances. PMID- 19545098 TI - Dual mechanism for type-2 diabetes resolution after Roux-en-Y gastric bypass. AB - Resolution of Type-2 diabetes mellitus (DM) after weight loss surgery is well documented, but the mechanism is elusive. We evaluated the glucose-insulin metabolism of patients undergoing a Roux-en-Y gastric bypass (RYGB) using the intravenous glucose tolerance test (IVGTT) and compared it with patients who underwent laparoscopic adjustable gastric band (AB) placement. Thirty-one female patients (age range, 20 to 50 years; body mass index, 47.2 kg/m2) underwent RYGB. Nine female patients underwent AB placement and served as control subjects. All patients underwent IVGTT at baseline and 1 month and 6 months after surgery. Thirteen patients undergoing RYGB and one patient undergoing AB exhibited impaired glucose tolerance defined by the American Diabetes Association. By 6 months post surgery, diabetes was resolved in all but one patient undergoing RYGB and none of the patients undergoing AB. Patients with diabetes undergoing demonstrated increased insulin secretion and beta-cell responsiveness 1 month after surgery and continued this trend up to 6 months, whereas none of the patients undergoing AB had changes in beta-cell function. Both patients undergoing RYGB and those undergoing AB demonstrated significant weight loss (34.6 and 35.0 kg/m2, respectively) and improved insulin sensitivity at 6 months. RYGB ameliorates DM resolution in two phases: 1) early augmentation of beta cell function at 1 month; and 2) attenuation of peripheral insulin resistance at 6 months. Patients undergoing AB only exhibited reduction in peripheral insulin resistance at 6 months but no changes in insulin secretion. PMID- 19545099 TI - Progressive preoperative pneumoperitoneum for hernias with loss of domain. AB - Acting as a pneumatic tissue expander, progressive preoperative pneumoperitoneum (PPP) pressurizes the abdomen to prepare patients with giant hernias and loss of domain for hernia repair. We review our experience with PPP. Between 2006 and 2008, a prospective hernia database revealed nine patients who underwent PPP prior to hernia repair. Mean patient age was 54 years (41-68) and mean BMI was 31.3 kg/m2 (25.2-36.5). Patients had prophylactic vena cava filters and intraperitoneal catheters placed. Over a mean 22.4 days (7-64), patients received 7.6 (3-13) injections of air. PPP complications included death from pulmonary mycetoma (1), deep venous thrombosis and acute renal failure (1), port infection (1), kinked catheter requiring reoperation (1). Seven patients underwent successful hernia repair; open ventral (6) and laparoscopic inguinal (1). Defect size averaged 387 cm2 (110-980) with a mesh size of 420 cm (180-1200). Operative time averaged 256 minutes (175-330) with a mean blood loss of 157 ml (50-500). Post-hernia repair length of stay was 10.3 days (4-22). Hernia repair complications included ventricular tachycardia (1) and hernia recurrence (1). PPP has an acceptable risk, and for patients with large hernias and loss of domain, it may be a useful adjunct prior to definitive hernia repair. PMID- 19545100 TI - False-positive and subsequent false negative Tc-99m sestamibi scan in a case of mediastinal thymoma and hyperparathyroidism. PMID- 19545101 TI - Pulmonary hypertension: an unusual presentation of an iatrogenic hepatic arterioportal fistula and its successful resolution post-embolotherapy. PMID- 19545102 TI - Cardiac surgery after bleomycin therapy. PMID- 19545103 TI - Subhepatic abscess caused by retained appendicolith: a rare complication requiring surgical intervention. PMID- 19545104 TI - Calciphylaxis. PMID- 19545105 TI - Tubular carcinoid tumor: a rare cause of appendicitis. PMID- 19545106 TI - Mucoepidermoid carcinoma of the thyroid and the parotid glands. PMID- 19545107 TI - Lymphoepithelial cysts of the pancreas: a sheep in wolves clothing. PMID- 19545108 TI - Surviving right atrial rupture. PMID- 19545109 TI - Primary hepatoid adenocarcinoma of retroperitoneum. PMID- 19545110 TI - Rupture of a giant renal artery aneurysm. PMID- 19545111 TI - Photocatalytic generation of oxygen radicals by the water-soluble bacteriochlorophyll derivative WST11, noncovalently bound to serum albumin. AB - Light-induced radical generation is the hallmark of fundamental processes and many applications including photosynthesis and photodynamic therapy (PDT). In this manuscript, we present two novel observations made upon monitoring light induced generation of reactive oxygen species (ROS) in aqueous solutions by WST11, a water-soluble derivative of the photosynthetic pigment Bacteriochlorophyll a (Bchl). Using a host of complementary experimental techniques including time-resolved spectroscopy at the subpicosecond to the millisecond range, ESR spectroscopy, electrochemistry, spectroelectrochemistry, oximetry, and protein mass spectroscopy, we first show that in aqueous solutions WST11 generates only superoxide (O(2)(-*)) and hydroxyl (OH*) radicals with no detectable traces of singlet oxygen. Second, we show that WST11 makes a noncovalent complex with human serum albumin (HSA) and that this complex functions as a photocatalytic oxidoreductase at biologically relevant concentrations enabling approximately 15 cycles of electron transfer from the associated HSA protein to molecular oxygen in the solution. These findings rule out the paradigm that porphyrin and chlorophyll based PDT is mainly mediated by formation of singlet oxygen, particularly in vascular targeted photodynamic therapy (VTP) with sensitizers that undergo photoactivation during circulation in the plasma, like [Pd]-Bacteriopheophorbide (WST09, Tookad). At the same time, our findings open the way for new design paradigms of novel sensitizers, since O(2)( *) and OH* radicals are well-recognized precursors of important pathophysiological processes that can be activated for achieving tumor eradication. Moreover, the finding that promiscuous protein scaffolds become sinks for holes and electrons when holding light-activated pigments provides a new insight to the evolution and action mechanism of natural light activated oxidoreductases (such as photosynthetic reaction centers) and new guidelines for the preparation of synthetic-light converting machineries. PMID- 19545113 TI - Infrared spectra and density functional calculations of the SUO2 molecule. AB - Reactions of laser-ablated U atoms with SO(2) molecules gave the very stable U(VI) molecule, SUO(2), as the major product. Infrared absorptions for two new O=U=O stretching modes were observed in solid argon and neon. The band assignments were confirmed by appropriate (34)SO(2), S(18)O(2), and S(16,18)O(2) isotopic shifts. B3LYP and BPW91 density functional calculations were performed to determine molecular structure, vibrational frequencies, and isotopic shifts. The C(2v) structure is analogous to those computed for UO(3) and US(3). Minor products were identified as SUO, the SUO(2)(+) cation, and the (SO(2))(SUO(2)) adduct. PMID- 19545115 TI - Preparation and properties of dextran sulfate-lysozyme conjugate. AB - The purpose of this research was to conjugate lysozyme with dextran sulfate (DS) under mild Maillard reaction conditions and to investigate the functional properties of the resulting conjugate. The covalent attachment of DS to lysozyme was confirmed by SDS-PAGE and cation-exchange chromatography. Lytic activity of the conjugated and uncongugated lysozyme toward Micrococcus lysodeikticus was 40 and 71% that of native lysozyme, respectively. The lysozyme-DS conjugate was highly soluble at alkaline pH values and different temperatures (25, 40, and 60 degrees C). Moreover, the modified lysozyme exhibited increased heat stability, with better emulsion activity and emulsion stability than those of unmodified proteins. Lysozyme-DS conjugate showed antibacterial activity and significantly inhibited growth of Staphylococcus aureus and Escherichia coli in a dose dependent manner. Taken together, the result of this study demonstrated that conjugation might increase the applicability of lysozyme in different food products for improving functional properties. PMID- 19545112 TI - Glycoproteomic analysis of embryonic stem cells: identification of potential glycobiomarkers using lectin affinity chromatography of glycopeptides. AB - Numerous studies have recently focused on the identification of specific glycan biomarkers, given the important roles that protein linked glycans play, for example, during development and disease progression. The identification of protein glycobiomarkers, which are part of a very complex proteome, has involved the use of fractionation techniques such as lectin affinity chromatography. In this study, the glycoproteomic characterization of pluripotent murine embryonic stem cells (ES) and from ES cells that were differentiated into embroid bodies (EB) was performed using immobilized Concanavalin A (ConA). This procedure allowed the isolation of glycopeptides that express biantennary and hybrid N linked structures (ConA2 fraction) as well as high mannose glycans (ConA3 fraction) that were abundant in both ES and EB stages. A total of 293 unique N linked glycopeptide sequences (from 180 glycoproteins) were identified in the combined data sets from ES and EB cells. Of these glycopeptides, a total of 119 sequences were identified exclusively in only one of the lectin-bound fractions (24 in the ES-ConA2, 15 in the ES-ConA3, 16 in the EB-ConA2, and 64 in the EB ConA3). Results from this study allowed the identification of individual N glycosylation sites of proteins that express specific glycan types. The absence of some of these lectin-bound glycopeptides in a cell stage suggested that they were derived from proteins that were either expressed exclusively on a defined developmental stage or were expressed in both cell stages but carried the lectin bound oligosaccharides in only one of them. Therefore, these lectin-bound glycopeptides can be considered as stage-specific glycobiomarkers. PMID- 19545116 TI - Thermal degradation kinetics of anthocyanins from blood orange, blackberry, and roselle using the arrhenius, eyring, and ball models. AB - Anthocyanin stability was assessed over temperatures ranging from 30 to 90 degrees C for seven products: blood orange juice [Citrus sinensis (L.) Osbeck]; two tropical highland blackberry juices (Rubus adenotrichus Schlech.), one with high content and the other with low content of suspended insoluble solids (SIS); and four roselle extracts (Hibiscus sabdariffa L.). The blackberry juice showed the highest content of anthocyanins with 1.2 g/L (two times less in the roselle extracts and 12 times less in the blood orange juice). The rate constant for anthocyanin degradation and isothermal kinetic parameters were calculated according to three models: Arrhenius, Eyring, and Ball. Anthocyanins in blood orange juice presented the highest rate constant for degradation, followed by the blackberry juices and roselle extracts. Values of activation energies were 66 and 37 kJ/mol, respectively, for blood orange and blackberry and 47-61 kJ/mol for roselle extracts. For the blackberry juices, a high SIS content provided only slight protection for the anthocyanins. The increasing content of dissolved oxygen, from 0.5 to 8.5 g/L, did not significantly increase the rate constant. For both isothermal and nonisothermal treatments, all three models accurately predicted anthocyanin losses from different food matrices. PMID- 19545117 TI - Investigation of natural phosphatidylcholine sources: separation and identification by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS2) of molecular species. AB - This study is a contribution to the exploration of natural phospholipid (PL) sources rich in long-chain polyunsaturated fatty acids (LC-PUFAs) with nutritional interest. Phosphatidylcholines (PCs) were purified from total lipid extracts of different food matrices, and their molecular species were separated and identified by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS(2)). Fragmentation of lithiated adducts allowed for the identification of fatty acids linked to the glycerol backbone. Soy PC was particularly rich in species containing essential fatty acids, such as (18:2 18:2)PC (34.0%), (16:0-18:2)PC (20.8%), and (18:1-18:2)PC (16.3%). PC from animal sources (ox liver and egg yolk) contained major molecular species, such as (16:0 18:2)PC, (16:0-18:1)PC, (18:0-18:2)PC, or (18:0-18:1)PC. Finally, marine source (krill oil), which was particularly rich in (16:0-20:5)PC and (16:0-22:6)PC, appeared to be an interesting potential source for food supplementation with LC PUFA-PLs, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). PMID- 19545118 TI - Production of a recombinant type 1 antifreeze protein analogue by L. lactis and its applications on frozen meat and frozen dough. AB - In this study, a novel recombinant type I antifreeze protein analogue (rAFP) was produced and secreted by Lactococcus lactis, a food-grade microorganism of major commercial importance. Antifreeze proteins are potent cryogenic protection agents for the cryopreservation of food and pharmaceutical materials. A food-grade expression and fermentation system (BSE- and antibiotic-free) for the production and secretion of high levels of rAFP was developed. Lyophilized, crude rAFP produced by L. lactis was tested in a frozen meat and frozen dough processing model. The frozen meat treated with the antifreeze protein showed less drip loss, less protein loss, and a high score on juiciness by sensory evaluation. Frozen dough treated with the rAFP showed better fermentation capacity than untreated frozen dough. Breads baked from frozen dough treated with rAFP acquired the same consumer acceptance as fresh bread. PMID- 19545119 TI - Chitosan-alginate blended nanoparticles as carriers for the transmucosal delivery of macromolecules. AB - Nanoparticles intended for use in the transmucosal delivery of macromolecules were prepared by the ionic gelation of chitosan (CS) hydrochloride with pentasodium tripolyphosphate (TPP) and concomitant complexation with sodium alginate (ALG). The incorporation of a small proportion of ALG of increasing molecular weight (M(w); from 4 to 74 kDa) into the nanoparticles led to a monotonic increase in colloidal size from ~260 to ~525 nm. This increase in size was regarded as a consequence of the formation of gradually more expanded structures. Insulin, taken as a model peptide, was associated to CS-TPP-ALG nanoparticles with efficiencies in the range of ~41 to ~52%, irrespective of the M(w) of the ALG incorporated in the formulation. These CS-TPP-ALG nanoparticles exhibited a capacity to enhance the systemic absorption of insulin after nasal administration to conscious rabbits. Interestingly, it was observed that the duration of the hypoglycaemic response was affected by the ALG's M(w). Briefly, this work describes a new nanoparticulate composition of potential value for increasing nasal insulin absorption. PMID- 19545122 TI - Preparation of N-maleoylchitosan nanocapsules for loading and sustained release of felodipine. AB - An interfacial free radical polymerization method was developed to fabricate polysaccharide nanocapsules, in which poor water-soluble drug of felodipine could be effectively encapsulated with good stability during storage. Exemplified by the preparation of felodipine-loaded N-maleoylchitosan (NMCS) nanocapsules, a felodipine/chloroform mixture was dispersed in NMCS aqueous solution with the aid of a nonionic surface active agent. After charging initiator, the vinylated groups of NMCS were polymerized on the oil-water interface. As a result, felodipine was loaded into NMCS nanocapsule. The morphology and the size distribution of synthesized nanocapsules were characterized by field emission scanning electron microscopy (FESEM) and dynamic light scattering (DLS) techniques. The quantitative drug loading and sustained release behavior were investigated. The encapsulation efficiency and drug loading content were found to be strongly dependent on the feed felodipine concentration. The release dynamics showed strong correlation with the degree of maleoyl substitution and the feed NMCS concentration during the course of nanocapsules preparation. PMID- 19545120 TI - Structure-activity relationships and cancer-cell selective toxicity of novel inhibitors of glioma-associated oncogene homologue 1 (Gli1) mediated transcription. AB - We report novel inhibitors of Gli1-mediated transcription as potential anticancer agents. Focused chemical libraries were designed and assessed for inhibition of functional cell-based Gli1-mediated transcription and selective toxicity toward cancer cells. The SAR was revealed, and the selectivity of the lead compounds' inhibition of Gli1-mediated transcription over that of Gli2 was determined. Compound 63 (NMDA298-1), which inhibited Gli1-mediated transcription in C3H10T1/2 cells with an IC(50) of 6.9 muM, showed 3-fold selectivity for inhibiting transcription mediated by Gli1 over that by Gli2. Cell-viability assays were performed to evaluate the chemical library in a normal cell line and a panel of cancer cell lines with or without up-regulated expression of the Gli1 gene. These compounds decreased the viability of several cancer cell lines but were less active in the noncancerous BJ-hTERT cells. PMID- 19545123 TI - Synthesis and characterization of polyrotaxane-amino acid conjugates: a new synthetic pathway for amino-functionalized polyrotaxanes. AB - Novel polyrotaxane conjugates possessing side chains of protected amino acids, that is, N(alpha)-tert-butyloxycarbonylglycine (Boc-Gly) and N(alpha) benzyloxycarbonylglycine (Z-Gly), were successfully prepared via the carbonyldiimidazole-mediated esterification of hydroxyl groups in a polyrotaxane. The prepared conjugates were soluble in a wide variety of organic solvents, including N,N-dimethylacetamide, N,N-dimethylformamide, tetrahydrofuran, pyridine, and methanol. Thermogravimetric measurements of the conjugates showed three-step decomposition curves corresponding to the decompositions of the amino acid, poly(ethylene glycol) axis, and cyclodextrin rings. The Z-Gly-polyrotaxane conjugate showed a remarkable exotherm at 334 degrees C, together with a large weight loss. Treatment of the Boc-Gly-polyrotaxane conjugate with neat trifluoroacetic acid resulted in the complete removal of the Boc groups and gave a cationic polyrotaxane with many free primary amino groups, the amount of which was determined by colloidal titrations. PMID- 19545124 TI - Exploring the structural properties of simple aldehydes: a Monte Carlo and small angle X-ray scattering study. AB - The structure of simple linear alkanals from propanal to nonanal was studied utilizing configurational bias Monte Carlo (MC) simulations of the aldehydes modeled according to the transferable potential for phase equilibria-united atom force field (TraPPE-UA) and was compared to experimental small-angle X-ray scattering (SAXS) results. This was done by exploiting a recently developed approach for calculating the scattering intensities from theoretically obtained MC data by utilizing the Debye equation (Tomsic et al. J. Phys. Chem. B 2007, 111, 1738). Similar calculations were also performed utilizing a well-established approach based on the reciprocal lattice. Comparison of the calculated scattering data with the experimental SAXS results in the first instance revealed information on the molecular organization in simple aldehydes and in addition also served as a good structural test of the TraPPE-UA force field used to model the aldehydes studied. However, it turned out that such a structural test is a rather strict test for the model which otherwise showed good agreement with the experimental data from the thermodynamic point of view. PMID- 19545125 TI - Influence of nonadiabatic annealing on the morphology and molecular structure of PEDOT-PSS films. AB - The effect of nonadiabatic annealing on poly(3,4-ethylendioxythiophene) poly(styrenesulfonate) (PEDOT-PSS) thin films prepared on silicon substrate has been investigated by Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). The analysis indicates the formation of an annealing-induced doping in PEDOT structure, suggesting a modification of the polymer electronic structure and the formation of a PEDOT-rich film surface. PMID- 19545126 TI - Oxidative and reductive metabolism of tris(p-carboxyltetrathiaaryl)methyl radicals by liver microsomes. AB - Tris(p-carboxyltetrathiaaryl)methyl (TAM) radicals are particularly stable carbon centered free radicals that are used as contrast agents in NMR imaging and as probes for in vivo oximetry by electron paramagnetic resonance (EPR) imaging. However, nothing is known so far on the metabolism of these persistent radicals in mammals. This article describes the metabolism of two TAM radicals by rat, human, and pig liver microsomes. It shows that microsomal transformation of these free radicals leads to two major metabolites resulting from an oxidation or a reduction of the present compounds. The structures of these metabolites were completely established by 1H and 13C NMR spectroscopy, mass spectrometry, and comparison with authentic compounds. Under aerobic conditions, liver microsomes catalyzed the oxidative decarboxylation of TAM radicals by NADPH and O2 with formation of the corresponding quinone-methide products. This reaction was dependent on cytochromes P450 and cytochrome P450 reductase and greatly implied the involvement of superoxide. Under anaerobic conditions, these enzymes catalyzed the reduction of TAM radicals to the corresponding triarylmethanes. This reduction was strongly inhibited by O2. These metabolic transformations should be considered when using such TAM radicals for pO2 measurement by EPR imaging, especially in tissues in which fast oxidative (inflammation sites) or reductive (hypoxic tissues) metabolism could occur. PMID- 19545127 TI - Sequence-directed organization of beta-peptides in self-assembled monolayers. AB - The sequence-directed organization of self-assembled monolayers of amphiphilic beta-peptides adsorbed on gold surfaces is studied using Monte Carlo simulations. A phenomenological model is presented in which each (helical) molecule is represented by a rigid nanorod; side groups are placed at appropriate locations. This model can distinguish between globally amphiphilic (GA) and nonglobally amphiphilic (iso-GA) sequence isomers. The simulations show that the GA isomers have a high degree of orientational order that is not exhibited by the iso-GA isomers, which is consistent with experiment (Pomerantz et al. Chem. Mater. 2007, 19, 4436). The effect of surface coverage and relative strength of electrostatic, hydrophilic, and hydrophobic interactions on the self-assembly of beta-peptides is quantified. PMID- 19545128 TI - EaMEAD: Activation energy prediction of cytochrome P450 mediated metabolism with effective atomic descriptors. AB - In an effort to improve drug design and predictions for pharmacokinetics (PK), an empirical model was developed to predict the activation energies (Ea) of cytochrome P450 (CYP450) mediated metabolism. The model, EaMEAD (Activation energy of Metabolism reactions with Effective Atomic Descriptors), predicts the Ea of four major metabolic reactions of the CYP450 enzyme: aliphatic hydroxylation, N-dealkylation, O-dealkylation, and aromatic hydroxylation. To build and validate the empirical model, the E(a) values of the substrates with diverse chemical structures (394 metabolic sites for aliphatic hydroxylation, 27 metabolic sites for N-dealkylation, 9 metabolic sites for O-dealkylation, and 85 metabolic sites for aromatic hydroxylation) were calculated by AM1 molecular orbital (MO). Empirical equations, Quantitative Structure Activity Relationship (QSAR) models, were derived using effective atomic charge, effective atomic polarizability, and bond dipole moments of the substrates as descriptors. EaMEAD is shown to accurately predict Ea with a correlation coefficient (R) of 0.94 and root-mean-square error (RMSE, unit is kcal/mol) of 0.70 for aliphatic hydroxylation, N-dealkylation, and O-dealkylation, and R of 0.83 and RMSE of 0.80 for aromatic hydroxylation, respectively. Physical origin and the role of the effective atomic descriptors of the models are presented in detail. With this model, the Ea of the metabolism can be rapidly predicted without any experimental parameters or time-consuming QM calculation. Regioselectivity prediction with our model is presented in the case of CYP3A4 metabolism. The reliability and ease of use of this model will greatly facilitate early stage PK predictions and rational drug design. Moreover, the model can be applied to develop the Ea prediction model of various types of chemical reactions. PMID- 19545129 TI - Structure determination of La18W10O57. AB - We report the synthesis, structure determination, and structure analysis of La(18)W(10)O(57), which presents 52.65 WO(3) mol % in the binary system La(2)O(3) WO(3) in place of the previous reported compound: La(14)W(8)O(45), 53.3 WO(3) mol %. The structure has been determined from synchrotron and laboratory X-ray, neutron, and electron diffraction data. This tungstate crystallizes in the non centrosymmetric hexagonal space group (no. 190) P62c, with Z = 2, a = 9.0448(1) A, c = 32.6846(3) A, and a measured density of 7.28(3) g x cm(-3). The structure consists of [WO(6)] octahedra, isolated or sometimes sharing a face, and unusual tungsten [WO(6)] trigonal prisms. This compound presents a subcell c' = c/6 = 5.45 A. Polytypism has been observed concerning this compound. Transmission electronic microscopy, ionic conductivity, and thermal expansion have been explored. PMID- 19545131 TI - Photo-cross-linkable polymer micelles in hydrogen-bonding-built layer-by-layer films. AB - Photoactive micelles of a diblock copolymer composed of poly(ethylene oxide) and poly(7-(2-methacryloyloxyethoxy)-4-methylcoumarin) (PEO-b-PCMA) were layer-by layer assembled with poly(acrylic acid) (PAA) using hydrogen-bonding between the PEO corona of the micelles and the PAA chains (pH < 3). In addition to characterizing the assembly process using a number of techniques, the tunable photo-cross-linking of polymer micelles through dimerization of the coumarin groups was used to generate interesting functions for the multilayer film. On the one hand, the easy tuning of the photo-cross-linking density could be used to control the release rate of hydrophobic guest molecules loaded in the film. On the other hand, after chemical cross-linking of PAA to stabilize the film, the photo-cross-linking of the micelles could be used to restrict the dissolution of PEO-b-PCMA chains in a good organic solvent; this cross-linking-dependent extraction of polymer micelles was utilized to vary the porosity of the film. PMID- 19545130 TI - High resolution crystal structure of the methylcobalamin analogues ethylcobalamin and butylcobalamin by X-ray synchrotron diffraction. AB - The X-ray crystal structures of the methylcobalamin (MeCbl) analogues ethylcobalamin (EtCbl) and butylcobalamin (BuCbl) are reported. The X-ray crystal structures of EtCbl and BuCbl were obtained with some of the lowest crystallographic residuals ever achieved for cobalamins (R = 0.0468 and 0.0438, respectively). The Co-C bond distances for EtCbl and BuCbl are 2.023(2) and 2.028(4) A, whereas the Co-alpha-5,6-dimethylbenzimidazole (Co-N3B) bond distances were 2.232(1) and 2.244(1) A, respectively. Although EtCbl and BuCbl displayed a longer Co-N3B bond than that observed in the naturally occurring methylcobalamin, the orientation of the alpha-5,6-dimethylbenzimidazole moiety with respect to the corrin ring did not vary substantially among the structures. The lengthening of both Co-C and Co-N3B bonds in EtCbl and BuCbl can be attributed to the "inverse" trans influence exerted by the sigma-donating alkyl groups, typically observed in alkylcobalamins. Analysis of the molecular surface maps showed that the alkyl ligands in EtCbl and BuCbl are directed toward the hydrophobic side of the corrin ring. The corrin fold angles in EtCbl and BuCbl were determined to be 14.7 degrees and 13.1 degrees, respectively. A rough correlation exists between the corrin fold angle and the length of the Co-N3B bond, and both alkylcobalamins follow the same trend. PMID- 19545133 TI - Catalysis of mukaiyama aldol reactions by a tricyclic aluminum alkoxide Lewis acid. AB - The Mukayiama aldol reaction of aldehydes is efficiently accomplished with a low concentration of the dimeric alumatrane catalyst 2 at mild or subambient temperatures. Our protocol tolerates a wide variety of electron-rich, neutral, and deficient aryl, alkyl, and heterocyclic aldehydes. A wide variety of enol silyl ethers are also tolerated. An intermediate that was isolated provides mechanistic information regarding the role of dimeric 2 in the Mukaiyama aldol reaction. Experimental evidence is presented for the stronger Lewis acidity of 5 compared with F3B. PMID- 19545132 TI - Discovery and development of the covalent hydrates of trifluoromethylated pyrazoles as riboflavin synthase inhibitors with antibiotic activity against Mycobacterium tuberculosis. AB - A high-throughput screening (HTS) hit compound displayed moderate inhibition of Mycobacterium tuberculosis and Escherichia coli riboflavin synthases. The structure of the hit compound provided by the commercial vendor was reassigned as [3-(4-chlorophenyl)-5-hydroxy-5-(trifluoromethyl)-4,5-dihydro-1H-pyrazol-1-yl](o tolyl)methanone (18). The hit compound had a k(is) of 8.7 microM vs. M. tuberculosis riboflavin synthase and moderate antibiotic activity against both M. tuberculosis replicating phenotype and nonreplicating persistent phenotype. Molecular modeling studies suggest that two inhibitor molecules bind in the active site of the enzyme, and that the binding is stabilized by stacking between the benzene rings of two adjacent ligands. The most potent antibiotic in the series proved to be [5-(4-chlorophenyl)-5-hydroxy-3-(trifluoromethyl)-4,5-dihydro 1H-pyrazol-1-yl](m-tolyl)methanone (16), which displayed a minimum inhibitory concentration (MIC) of 36.6 microM vs. M. tuberculosis replicating phenotype and 48.9 microM vs. M. tuberculosis nonreplicating phenotype. The HTS hit compound and its analogues provide the first examples of riboflavin synthase inhibitors with antibiotic activity. PMID- 19545134 TI - Differential reactivity pattern of hybrid o-quinodimethane precursors: strategic expansion to annulated benzocycloalkanes via rongalite. AB - A hybrid benzocyclobutene (BCB) molecular frames embedding sultine or sulfone moiety has been synthesized via utilization of rongalite. The selective Diels Alder reaction has been realized at sultine or sulfone terminus in the hybrid BCB system to prepare functionalized BCB molecular frames. The methodology has been generalized for assembling various benzocycloalkanes containing a sultine unit and the strategy has been expanded to generate various annulated benzocycloalkanes. PMID- 19545135 TI - Synthesis of small atomic copper clusters in microemulsions. AB - We report evidence of the formation of small atomic copper clusters, Cu(n), by the microemulsion technique, and how their size can be controlled by adjusting the percentage of the reducing agent used. Copper clusters were characterized by UV-visible spectrophotometry and atomic force microscopy. Photoluminescent copper clusters, Cu(n), with n less, similar 13, can be obtained using very low percentages of the reducing agent (<10% of the stochiometric amount). Photoluminescent clusters disappear for larger percentages of reducing agent, giving rise to larger copper clusters (0.8-2.0 nm), showing a red-shift of their UV-visible absorption bands as they grow in size. Finally, by using near stoichiometric amounts, nanoparticles of 2.9 +/- 1.1 nm in size, displaying the characteristic plasmon band, can be obtained. PMID- 19545136 TI - Performance of SCS palladium pincer complexes in borylation of allylic alcohols. Control of the regioselectivity in the one-pot borylation-allylation process. AB - One-pot borylation-allylation reactions of aldehydes and allylic alcohols were performed under various reaction conditions. The borylation of allylic alcohols was performed using a very efficient SCS palladium pincer-complex catalyst. The regioselectivity of the allylation depends on the applied solvent. The reaction in CHCl3 gave the linear allylic product; however, when MeOH was added to the reaction mixture, the branched allylic product was formed. PMID- 19545137 TI - Focused ion beam fabrication of boron-doped diamond ultramicroelectrodes. AB - The fabrication of ultramicroelectrodes (UMEs) for analytical electrochemical applications has been explored, using boron-doped diamond as the active electrode material in an insulating coating formed by deposition of electrophoretic paint. Because of the rough nature of the diamond film, the property of such coatings that is normally exploited in the fabrication of UMEs, namely the tendency to retract automatically from sharp protrusions, cannot be used in the present instance. Instead focused ion beam (FIB) sputtering was employed to controllably produce UMEs with well-defined geometry, critical dimension of a few micrometers, and very thin insulating coatings. If the FIB machining is carried out at normal incidence to the diamond electrode surface, significant ion beam damage reduces the yield of successful electrodes. However, if a parallel machining geometry is employed, high yields of ultramicroelectrodes with a flat disk geometry can be obtained very reliably. The electrochemical properties of diamond UMEs are characterized. They show much lower background currents than the equivalent Pt or carbon fiber electrodes but more varied electrochemical response than macroscopic diamond electrodes. PMID- 19545138 TI - Electrochemistry of the inclusion complexes formed between the cucurbit[7]uril host and several cationic and neutral ferrocene derivatives. AB - We have investigated the formation of inclusion complexes between the host cucurbit[7]uril (CB7) and three cationic and four neutral ferrocene-containing guests: (ferrocenylmethyl)trimethylammonium (2(+)), butyl(ferrocenylmethyl) dimethylammonium (3(+)), (ferrocenylmethyl)heptyldimethylammonium (4(+)), hydroxymethylferrocene (5), (((methoxy)-ethoxy)ethoxy)methylferrocene (6), 1,1' di(hydroxymethyl)ferrocene (7), and 1,1' di((((methoxy)ethoxy)ethoxy)methyl)ferrocene (8). The formation of highly stable inclusion complexes (K > 10(7) M(-1)) was verified in all cases using NMR spectroscopic techniques. From cyclic voltammetric experiments, we observed that CB7 complexation of the cationic guests (2(+)-4(+)) leads to significant anodic shifts on the ferrocene oxidation half-wave potentials, while the measured potential shifts were smaller in the case of the neutral guests (5-8). Encapsulation of all guests resulted in a substantial decrease of the standard rate constant for heterogeneous electron transfer. However, inclusion complexation of the neutral guests led to quasi-reversible voltammetric behavior, in which the anodic peak potential is more sensitive to scan rate than the corresponding cathodic peak potential, suggesting a minor degree of structural rearrangement in the neutral inclusion complex after oxidation. PMID- 19545140 TI - One-pot synthesis of gold nanorods by ultrasonic irradiation: the effect of pH on the shape of the gold nanorods and nanoparticles. AB - A novel one-pot synthesis method to prepare gold nanorods was developed by using sonochemical reduction of gold ions in aqueous solution. The size of the sonochemically formed gold nanorods was less than 50 nm, and their average aspect ratio decreased with increasing pH of the solution. The aspect ratio measured was 3.0 at pH 3.5, 2.2 at pH 5.0, and 2.1 at pH 6.5. At pH 7.7, irregular shaped gold nanoparticles were formed. At pH 9.8, most of the particles formed had a spherical shape with a smaller particle size than those formed in the lower pH solutions. Based on the obtained results, it was clear that the size and shape of the sonochemically formed gold nanoparticles are dramatically dependent on the pH value of the solution. PMID- 19545139 TI - Oxidation of a water-soluble phosphine and some spectroscopic probes with nitric oxide and nitrous acid in aqueous solutions. AB - In acidic aqueous solutions, nitrogen monoxide oxidizes monosulfonated triphenylphosphine, TPPMS(-), to the corresponding phosphine oxide. The NO derived product is N(2)O. This chemistry parallels that reported for the reaction of NO with the unsubstituted triphenylphosphine in nonpolar organic solvents, but the rate constant measured in this work, 5.14 x 10(6) M(-2) s(-1), is greater by several orders of magnitude. This makes TPPMS(-) a useful analytical reagent for NO in aqueous solution. The increased rate constant in the present work appears to be a medium effect, and unrelated to the introduction of a single sulfonate group in the phosphine. The reaction between nitrous acid and TPPMS(-) has a 2:1 [TPPMS(-)]/[HNO(2)] stoichiometry and generates NH(2)OH quantitatively. The rate law, rate = 4k(d)[HNO(2)](2)[TPPMS(-)], identifies the second-order self-reaction of HNO(2) as the rate-limiting step that generates the active oxidant(s) for the fast subsequent reaction with TPPMS(-). It appears that the active oxidant is N(2)O(3), although the oxides NO and NO(2) derived from it may be also involved. Bimolecular self-reaction of HNO(2) also precedes the oxidations of ABTS(2-) and TMPD. Competing with this path are the acid-catalyzed oxidations of both reagents via NO(+). PMID- 19545141 TI - Poly[perfluorotitanate(IV)] salts of [H3O]+, Cs+, [Me4N]+, and [Ph4P]+ and about the existence of an isolated [Ti2F9]- anion in the solid state. AB - The increase in the size of monocations (A(+)) does not favor the formation of [Ti(2)F(9)](-) against [Ti(4)F(18)](2-) salts (with isolated [Ti(2)F(9)](-) or [Ti(4)F(18)](2-) anions, respectively) as previously proposed (Passmore, J.; et al. Angew. Chem., Int. Ed. 2005, 44, 7958-7961). The crystal structure determination of [Me(4)N](+) and [Ph(4)P](+) salts showed that both compounds are [Ti(4)F(18)](2-) salts; i.e., [Me(4)N](2)[Ti(4)F(18)] and [Ph(4)P](2)[Ti(4)F(18)] were obtained instead of [Me(4)N][Ti(2)F(9)] and [Ph(4)P][Ti(2)F(9)]. The product of the reaction of CsF with 2TiF(4) could be formulated as CsTi(2)F(9); however, instead of isolated [Ti(2)F(9)](-) anions, infinite ([Ti(2)F(9)](-))(n) double chains are present. In the case of H(3)OTi(2)F(9), a similar result was obtained. On the basis of the similarities of vibrational spectra of CsTi(2)F(9) and NF(4)Ti(2)F(9), it is also unlikely that the latter consists of isolated [Ti(2)F(9)](-) ions. PMID- 19545142 TI - NO-Induced Reorganization of Porphyrin Arrays. AB - We studied the interaction of a highly ordered array of Co-tetraphenylporphyrin (CoTPP) with NO on Ag(111) by in situ scanning tunneling microscopy and X-ray photoelectron spectroscopy. Upon NO exposure, the initially quadratically ordered CoTPP layer reorganizes, showing a wealth of highly ordered NO+CoTPP coadsorbate phases with increasing size of the unit cell, interpreted as due to attractive lateral dipole-dipole interactions between the two species. The findings not only suggest a novel approach to control the arrangement of adsorbed porphyrins in particular but also should generally be considered in the production of functional layers from large organic molecules under ambient conditions or after exposure to small electronegative molecules. PMID- 19545144 TI - Single-crystal-to-single-crystal transformation in unusual three-dimensional manganese(II) frameworks exhibiting unprecedented topology and homospin ferrimagnet. AB - A new three-dimensional Mn(II) complex, [Mn(3)(HCO(2))(2)(L)(2)(OCH(3))(2)], 1 (L = nicotinate N-oxide), was synthesized by solvothermal reaction and magnetically characterized. Complex 1 exhibits an unprecedented 3,6-connected 5-nodal net topology with Schlafli notation {4(2);6}4{4(3)}2{4(6);6(6);8(3);10}2{4(8);6(6);8} and was assigned as a homospin ferrimagnet. Interestingly, when 1 was placed in air for ca. one month, the methoxy anions in 1 were gradually exchanged by hydroxyl anions and 1 underwent a single-crystal-to-single-crystal structural transition to a new but similar complex, [Mn(3)(HCO(2))(2)(L)(2)(OH)(2)] x 4 H(2)O, 2. The anion exchange and water molecules filling the channels of 2 affect the magnetic behavior at low temperature compared to that of 1. PMID- 19545145 TI - Biomimetic synthesis of a new class of bacterial signaling molecules. AB - The first synthesis of a newly discovered class of bacterial signaling molecules from Streptomyces coelicolor has been developed. These molecules, known as the methylenomycin furans (MMFs), trigger production of the antibiotic methylenomycin. The synthesis features a scandium triflate-catalyzed domino reaction of beta-ketoesters and dihydroxyacetone yielding 2,3,4-substituted furans. The proposed reaction sequence (aldol reaction, cyclization, and dehydrative aromatization) may be reminiscent of the biosynthetic reaction in which dihydroxyacetone phosphate and a beta-ketothioester are condensed by an enzyme. PMID- 19545143 TI - An indole-containing dauer pheromone component with unusual dauer inhibitory activity at higher concentrations. AB - In Caenorhabditis elegans, the dauer pheromone, which consists of a number of derivatives of the 3,6-dideoxysugar ascarylose, is the primary cue for entry into the stress-resistant, "nonaging" dauer larval stage. Here, using activity-guided fractionation and NMR-based structure elucidation, a structurally novel, indole-3 carboxyl-modified ascaroside is identified that promotes dauer formation at low nanomolar concentrations but inhibits dauer formation at higher concentrations. PMID- 19545146 TI - Homometallic uranium(VI) phosphonoacetates containing interlayer dipyridines. AB - Three organically templated uranium(VI) carboxyphosphonates, [C(10)H(10)N(2)][(UO(2))(3)(PPA)(2)(HPPA)(H(2)O)] x 3 H(2)O (1), [C(12)H(12)N(2)][(UO(2))(3)(PPA)(2)(HPPA)(H(2)O)] x 2 H(2)O (2), and [C(12)H(14)N(2)][(UO(2))(3)(PPA)(2)(HPPA)(H(2)O)] x 2 H(2)O (3) have been prepared from hydrothermal reactions of UO(2)(NO(3))(2) x 6 H(2)O and phosphonoacetic acid (H(2)O(3)PCH(2)CO(2)H, H(3)PPA) in the presence of 4,4' dipyridyl, 1,2-bis(4-pyridyl)-ethylene and 1,2-bis(4-pyridyl)-ethane. The crystal structures of 1-3 are built from UO(7) units linked via phosphonoacetate molecules to form 2-dimensional [(UO(2))(3)(PPA)(2)(HPPA)(H(2)O)](2-) networks which are charge balanced by doubly protonated bipyridyl molecules. Interaction of the pyridyl molecules with other functionalities present in 1-3 result in relatively strong N-H...O (2.707-3.155 A) and weaker C-H...O (2.948-3.237 A) interactions that constitute the interlayer. Water-carboxylate O-H...O hydrogen bonding (2.548-2.675 A) in 1-3 and pi-pi interactions (3.708 A) between pyridyl rings in 2 also exist in these materials. Reported herein are the syntheses, structural characterization, and thermal and fluorescent behavior of 1-3. PMID- 19545147 TI - Targeting the DNA damage response in cancer. PMID- 19545148 TI - 3,4-Dihydroxyphenylglycol (DHPG): an important phenolic compound present in natural table olives. AB - The presence of 3,4-dihydroxyphenylglycol (DHPG) was studied in 32 samples and 10 different cultivars of natural table olives, using an accurate method to avoid wrong quantification. Hydroxytyrosol (HT), tyrosol, and verbascoside were also quantified, as these four compounds comprise the majority of the chromatographic profile. Analyses were carried out by HPLC-DAD-UV after extraction of all phenolics, and hydroxytyrosol was the major component in nearly all samples. High levels of DHPG (up to 368 mg/kg of dry weight) were found in the pulp of natural black olives independent of cultivar and processing method, similar to its concentration in the brine in almost all of the samples. The presented data for this antioxidant indicate that natural table olives are a rich source of DHPG and hydroxytyrosol, compounds with interesting nutritional and antioxidant properties. PMID- 19545150 TI - Quantitative PCR method to measure the fungal endophyte in locoweeds. AB - A fungal endophyte ( Undifilum oxytropis ) has been implicated in the synthesis of swainsonine in Oxytropis and Astragalus species, commonly known as locoweeds. A quantitative PCR method has been developed to measure the amount of endophyte in Oxytropis and Astragalus species. The limit of quantitation was estimated to be 0.2 pg of endophyte/ng of total DNA. This method of analysis was used to quantify the amount of endophyte in 10 plants each of Oxytropis sericea (white point locoweed), Astragalus mollissimus (wooly locoweed), and Astragalus lentiginosus (spotted locoweed). A significant amount of individual plant variability was observed in endophyte content among individuals in all three species. In one O. sericea and one A. lentiginosus plant swainsonine concentrations were near or below the limit of detection. These plants also had the lowest amounts of endophyte when compared to the other specimens. This method will be a useful tool in further investigating the role the endophyte plays in swainsonine production in various locoweed species. PMID- 19545149 TI - Rapid label-free identification of estrogen-induced differential protein expression in vivo from mouse brain and uterine tissue. AB - Protein abundance profiling from tissue using liquid chromatography-tandem mass spectrometry-based "shotgun" proteomics and label-free relative quantitation was evaluated for the investigation of estrogen-regulated protein expression in the mouse brain and uterus. Sample preparation involved a 30-min protein extraction in 8 M aqueous urea solution, followed by disulfide reduction, thiol alkylation, and trypsin digestion of the extracted proteins, and was performed on 3-4 mg of tissue to evaluate the suitability of this methodology to expedite the survey of cellular pathways that are affected in vivo by an experimental therapeutic intervention in an animal model. The label-free proteomic approach (spectral counting) was suitable to identify even subtle changes in cortical protein levels and revealed significant estrogen-induced upregulation of ATP synthase (both alpha- and beta-isoforms), aspartate aminotransferase 2, and mitochondrial malate dehydrogenase without any prior subcellular fractionation of the tissue or the use of multidimensional chromatographic separation. The methodology was also suitable to observe various up- and downregulated proteins in the uterine tissue of ovariectomized mice upon treatment with 17beta-estradiol. In addition to confirming a very significant decrease in the abundance of glutathione S transferase recognized as a marker of estrogen's impact, our studies have also revealed potential new protein markers such as desmin and lumican that are critical components of cytoskeletal arrangement and, hence, regulation of their abundance could contribute to major morphological changes in the uterus occurring upon estrogenic stimulation. PMID- 19545151 TI - NMR metabolic profiling of transgenic maize with the Cry1Ab gene. AB - The metabolic profiles of seeds from the transgenic maize variety 33P67 and of the corresponding traditional variety were investigated using one- and two dimensional NMR techniques. The transgenic variety carries a functional Cry1A(b) gene, which confers to the plant the ability to produce Bt insect toxin. About 40 water-soluble metabolites in the maize seed extracts were identified, providing a more complete (1)H and (13)C NMR assignment with respect to the assignment reported in the literature. In particular ethanol, lactic acid, citric acid, lysine, arginine, glycine-betaine, raffinose, trehalose, alpha-galactose, and adenine were identified for the first time in the (1)H NMR spectrum of maize seeds extracts. The (1)H spectra of transgenic and nontransgenic seed maize samples turned out to be conservative, showing the same signals and therefore the same metabolites. However, a higher concentration of ethanol, citric acid, glycine-betaine, trehalose, as well as of another compound not yet completely identified, was observed in the transgenic extracts than in nontransgenic samples. So, it was possible to discriminate between transgenic and nontransgenic metabolic profilings through the use of an appropriate statistical analysis. PMID- 19545152 TI - BaNbO(IO3)5: a new polar material with a very large SHG response. AB - By combination of Nb(5+) (having a d(0) electronic configuration) and the lone pair-containing iodate anion, a new SHG material, BaNbO(IO(3))(5), has been prepared. It exhibits a very large SHG response (approximately 14 times that of KH(2)PO(4) and approximately 660 times that of alpha-SiO(2)) and is phase matchable. The material has high thermal stability and a wide transparent region. PMID- 19545153 TI - Palladium-catalyzed carboamination of alkenes promoted by N fluorobenzenesulfonimide via C-H activation of arenes. AB - This report describes a unique Pd-catalyzed oxidative carboamination of protected aminoalkenes in which inexpensive unactivated nucleophilic arenes are incorporated to give carboamination products in good yields. A variety of protected amide and carbamate groups are tolerated, and various five-, six-, and seven-membered rings are formed in good yields. Under these conditions, halobenzenes are activated at the C-H bond rather than the C-X bond, and very high regioselectivity for the para substitution product is observed in all cases. We propose that this carboamination takes place via electrophilic aromatic substitution of a Pd(IV) alkyl intermediate. PMID- 19545154 TI - Identifying apoptosis-evasion proteins/pathways in human hepatoma cells via induction of cellular hormesis by UV irradiation. AB - Evading apoptosis is pivotal in both of carcinogenesis and resistance to anticancer therapy. We investigated the molecules and pathways of apoptosis evasion in human hepatoma cells by irradiating hepatoma cells with optimized UV (so-called "hormetic responses"). Proteins and pathways related to hormetic responses were identified via proteomic approaches followed by reconstruction of function-networks. Of the 2326 defined protein spots, 42 distinct proteins significantly changed their expression. Eleven hormetic response proteins (HINT1, PHB, CTSD, ANXA1, LGASL1, TPT1, NPM, PRDX2, UCHL1, CERK, and C1QBP) were involved in 5 death-regulatory pathways, including the p53-dependent apoptotic pathway, protein ubiquinization, cellular redox, calcium-mediated signaling pathway, and sphingomyelin-metabolism pathway. Knockdown of HINT1 expression via RNA interference increased tumor cell resistance to apoptosis induction, while silencing NPM, UCHL1, or CERK greatly sensitized tumor cells to apoptosis induction. In conclusion, NPM, UCHL1, and CERK act as apoptosis-evasion proteins that may serve as therapeutic targets for hepatoma. Silencing their expression would increase therapeutic efficacy, thereby reducing the corresponding doses and side-effects of anticancer therapy. This model of induction of cellular hormetic responses to identify apoptosis-evasion molecules/pathways via proteomic approaches can be applied to other modalities of anticancer therapy. PMID- 19545155 TI - Electronic structure, binding energy, and solvation structure of the streptavidin biotin supramolecular complex: ONIOM and 3D-RISM study. AB - We studied the electronic structure of the binding site of the streptavidin biotin complex by using the ONIOM method at the HF/STO-3G:UFF level and obtained the solvation structure of the complex by using the statistical-mechanical, three dimensional molecular theory of solvation (aka three-dimensional reference interaction site model, 3D-RISM-KH). All the streptavidin residues located within 3 A of the biotin residue were included in the quantum mechanical (QM) layer. In total, 16 residues including biotin with 274 atoms were in the QM layer, in which five residues are responsible for the hydrophobic interactions and nine residues for the hydrogen-bonding/electrostatic interaction with biotin. We found a geometry change of the urea moiety of the biotin bound in the network of van der Waals and polar interactions. Compared to the isolated biotin, the bridging C-C bond of the biotin urea moiety in the binding site increases in length as a result of the pi-sigma interaction with the surrounding streptavidin Trp residues. This extends the previous picture of the geometry change from the ureido group to the whole bicyclic urea moiety. We have evaluated the performance of 15 density functional methods and 11 basis sets by single point calculation for the binding energy of the optimized cooperative binding complex structure. Closest to the experimental value of 18.3 kcal/mol is the binding free energy of 19.6 kcal/mol obtained for the AN model at B3LYP/6-31G(d):UFF//HF/STO-3G:UFF level. The hybrid DFT methods with enhanced assessment for nonbonded interactions such as PBE1PBE, MPW1B95, and MPWB1K can also give accurate binding energy with the use of diffuse functionals (i.e., mPWB1K/6-31+G(d)). The 3D hydration structure of the unliganded streptavidin and the streptavidin-biotin complex obtained by using the 3D-RISM-KH molecular theory of solvation shows there is one immobilized water molecule at the biotin urea moiety, acting as a water bridge between the sulfur and the nitrogen of the NH group close to Ser45. This suggests that, in the docking process, biotin replaces six of the seven water molecules attached to the unliganded streptavidin binding site, and one remaining water molecule is squeezed into the gap between the Btn, Tyr43, Ser45, Trp92, and Trp79 residues in the binding pocket. PMID- 19545156 TI - Poly(3,4-ethylenedioxythiophene)-multiwalled carbon nanotube composite films: structure-directed amplified electrochromic response and improved redox activity. AB - Composite thin films of poly(3,4-ethylenedioxythiophene) (PEDOT)-enwrapped functionalized multiwalled carbon nanotubes (MWCNTs) have been synthesized over multiple length scales by electropolymerization of the monomer without the use of any other supporting electrolyte. The functionalized MWCNTs are incorporated into the positively charged polymer deposit as counterions during oxidative electropolymerization. The morphology, electrochemistry, and electrochromism of the PEDOT-MWCNT films have been compared with those of control PEDOT films doped by triflate ions. Such a comparison enabled us to demonstrate the profound effect of MWCNTs as counterions, realized in terms of better electropolymerization rate, higher conductivity, faster color-bleach kinetics, higher charge storage capacity, and substantially amplified coloration efficiency (eta = 414 cm(2) C( 1), lambda(max) = 575 nm, E = -1.5 V) in comparison to the values of eta reported to date for PEDOT. The strong interaction between the polymer and MWCNTs, the interconnected nanotubular structures, and the porous framework of the film allow facile charge transport and larger ion uptake during redox switching. Electrochemical investigations on devices based on PEDOT-MWCNT and control PEDOT films established the practical utility of PEDOT-MWCNT films as they show lower charge-transfer resistance, higher diffusional capacitance, and a much smaller amplitude of impedance as compared to control PEDOT films. PMID- 19545158 TI - Nuclear magnetic resonance signaling of molecular chiral information using an achiral reagent. AB - Until now NMR spectroscopic detection of guest chirality using an achiral host has not been possible in the absence of a chiral medium or auxiliary since chiral discrimination is principally based on chiral discrimination by host and/or diastereomeric host-guest complex formation. In this paper, we demonstrate that an achiral oxoporphyrinogen works as a host capable of signaling chiral information of alpha-hydroxycarboxylic acids in (1)H NMR spectroscopy. In particular, enantiomeric excess (ee) can be determined by observing the splitting of (1)H NMR resonances of the achiral host. This differs from the case of chiral hosts (shift reagents) where % ee is generally determined from the ratio of peak areas due to diastereomeric host-guest complexes. UV/vis, CD, FT-IR, and NMR spectroscopic investigations suggest that the unusual phenomenon reported here is based on formation of a complex with 1:2 stoichiometry in concert with a protonation-driven tautomerization of the host. PMID- 19545159 TI - Quaternary alloy semiconductor nanobelts with bandgap spanning the entire visible spectrum. AB - We used an improved cothermal evaporation route for the first time to achieve quaternary semiconductor nanostructured alloys, using an example of Zn(x)Cd(1 x)S(y)Se(1-y) nanobelts. The PL (bandgap) of these as-grown nanostructured alloys can be continuously tunable across the entire visible spectrum through experimentally controlling their compositions. Such widely controlled alloy nanostructures via composition/light emission provide a new material platform for applications in wavelength-tunable lasers, multicolor detectors, full-spectrum solar cells, LEDs, and color displays. PMID- 19545157 TI - Novel autoimmune hepatitis-specific autoantigens identified using protein microarray technology. AB - Autoimmune hepatitis (AIH) is a chronic necroinflammatory disease of the liver with a poorly understood etiology. Detection of nonorgan-specific and liver related autoantibodies using immunoserological approaches has been widely used for diagnosis and prognosis. However, unambiguous and accurate detection of the disease requires the identification and characterization of disease-specific autoantigens. In the present study, we have profiled the autoantigen repertoire of patients with AIH versus those with other liver diseases, identifying and validating three novel and highly specific biomarkers for AIH. In phase I, we fabricated a human protein chip of 5011 nonredundant proteins and used it to quickly identify 11 candidate autoantigens with relative small serum collection. In phase II, we fabricated an AIH-specific protein chip and obtained autoimmunogenic profiles of serum samples from 44 AIH patients, 50 healthy controls, and 184 additional patients suffering from hepatitis B, hepatitis C, systemic lupus erythematosus, primary Sjogren's syndrome, rheumatoid arthritis, or primary biliary cirrhosis. With this two-phase approach, we identified three new antigens, RPS20, Alba-like, and dUTPase, as highly AIH-specific biomarkers, with sensitivities of 47.5% (RPS20), 45.5% (Alba-like), and 22.7% (dUTPase). These potential biomarkers were further validated with additional AIH samples in a double-blind design. Finally, we demonstrated that these new biomarkers could be readily applied to ELISA-based assays for use in clinical diagnosis/prognosis. PMID- 19545160 TI - Propulsion of gold nanoparticles with surface plasmon polaritons: evidence of enhanced optical force from near-field coupling between gold particle and gold film. AB - We experimentally demonstrate the enhanced propulsion of gold nanoparticles by surface plasmon polaritons (SPPs). Three dimensional finite difference time domain (FDTD) simulations indicate considerably enhanced optical forces due to the field enhancement provided by SPPs and the near-field coupling between the gold particles and the film. This coupling is an important part of the enhanced propulsion phenomenon. Finally, the measured optical force is compared with that predicted by FDTD simulations and proven to be reasonable. PMID- 19545161 TI - Deciphering the structural properties that confer stability to a DNA nanocage. AB - A DNA nanocage has been recently characterized by small-angle X-ray scattering (SAXS) and cryo-transmission electron microscopy as a DNA octahedron having a central cavity larger than the apertures in the surrounding DNA lattice. Starting from the SAXS data, a DNA nanocage has been modeled and simulated by classical molecular dynamics to evaluate in silico its structural properties and stability. Global properties, principal component analysis, and DNA geometrical parameters, calculated along the entire trajectory, indicate that the cage is stable and that the B-DNA conformation, also if slightly distorted, is maintained for all the simulation time. Starting from the initial model, the nanocage scaffold undergoes a contraction of the thymidine strands, connecting the DNA double helices, suggesting that the length of the thymidine strands is a crucial aspect in the modulation of the nanocage stability. A comparison of the average structure as obtained from the simulation shows good agreement with the SAXS experimental data. PMID- 19545162 TI - Synthesis of prostaglandin and phytoprostane B1 via regioselective intermolecular Pauson-Khand reactions. AB - A new approach to the synthesis of prostaglandin and phytoprostanes B(1) is described. The key step is an intermolecular Pauson-Khand reaction between a silyl-protected propargyl acetylene and ethylene. This reaction, promoted by NMO in the presence of 4 A molecular sieves, afforded the 3-tert butyldimethylsilyloxymethyl-2-substituted-cyclopent-2-en-1-ones (III) in good yield and with complete regioselectivity. Deprotection of the silyl ether, followed by Swern oxidation, gave 3-formyl-2-substituted-cyclopent-2-en-1-ones (II). Julia olefination of the aldehydes II with the suitable chiral sulfone enabled preparation of PPB(1) type I and PGB(1). PMID- 19545164 TI - The collective behavior of nanoscale building blocks. AB - Nanotechnology has reached the stage of development where the subject of most investigations is not individual nanoparticles or nanowires but rather systems of much greater complexity. PMID- 19545163 TI - Water-soluble rhamnose-coated Fe3O4 nanoparticles. AB - Water-soluble biocompatible rhamnose-coated Fe(3)O(4) nanoparticles of 4.0 nm are obtained by covalent anchorage of rhamnose on the nanoparticles surface via a phosphate linker. These nanoparticles present superparamagnetic behavior and nuclear relaxivities in the same order of magnitude as Endorem that make them potential magnetic resonance imaging (MRI) contrast agents of a second generation, where the saccharides represent also specific ligands able to target lectins on skin cells. PMID- 19545166 TI - A conversation with Prof. Chad Mirkin: nanomaterials architect. PMID- 19545167 TI - Characterization of antiplatelet properties of silver nanoparticles. AB - Thrombotic disorders have emerged as serious threat to society. As anticoagulant and thrombolytic therapies are usually associated with serious bleeding complications, the focus has now shifted to regulating and maintaining platelets in an inactive state. In the present study we show that nanosilver has an innate antiplatelet property and effectively prevents integrin-mediated platelet responses, both in vivo and in vitro, in a concentration-dependent manner. Ultrastructural studies show that nanosilver accumulates within platelet granules and reduces interplatelet proximity. Our findings further suggest that these nanoparticles do not confer any lytic effect on platelets and thus hold potential to be promoted as antiplatelet/antithrombotic agents after careful evaluation of toxic effects. PMID- 19545168 TI - Self-diffusion of water and simple alcohols in single-walled aluminosilicate nanotubes. AB - Understanding transport phenomena of fluids through nanotubes (NTs) is of great interest in order to enable potential application of NTs as separation devices, encapsulation media for molecule storage and delivery, and sensors. Single-walled metal oxide NTs are interesting materials because they present a well-defined solid-state structure, precisely tunable diameter and length, as well as a hydrophilic and functionalizable interior for tuning transport and adsorption selectivity. Here, we study the transport properties of hydrogen-bonding liquids (water, methanol, and ethanol) through a single-walled aluminosilicate NT to investigate the influence of liquid-surface and liquid-liquid interactions and the effects of competitive transport of different chemical species using molecular dynamics (MD) simulations. The self-diffusivities (D(s)) for all the three species decrease with increasing loading and are comparable to bulk liquid diffusivities at low molecular loadings. We show that the hydrogen-bond network associated with water makes its diffusion behavior different from methanol and ethanol. Mixtures of water and methanol show segregation in the NT, with water located closer to the tube wall and the alcohol molecules localized near the center of the NT. D(s) values of water in an analogous aluminogermanate NT are larger than those in the aluminosilicate NT due to a larger pore diameter. PMID- 19545169 TI - Zn cluster drifting effect for the formation of ZnO 3D nanoarchitecture. AB - Metal catalysts are widely used for nanowire (NW) growth and are one of the essential parameters that dictate the crystal growth phenomena, thus controlling the NW's morphology. Although extensive research has been conducted on catalyst effects, the catalyst drifting effect is generally underestimated for controlling the morphology of nanostructures grown at a relatively high temperature. In this paper, we report a discovery of Zn cluster drifting phenomenon during ZnO vapor deposition. Because of the deposition of ZnO along the drifting path, the dynamic process of cluster drifting could be visualized after the growth. This phenomenon provides a sound explanation of the formation of randomly orientated ZnO nanowall networks. The cluster drifting direction could be intentionally directed by designing the surface inclination, through which a partially parallel aligned ZnO vertical nanofin array was created. This 3D nanoarchitecture would possibly provide a novel configuration for designing high performance integrated nanodevices. The drifting of Zn clusters could be a general phenomenon for most metal catalysts and would provide a new insight into nanofabrication and nanodevice development. PMID- 19545170 TI - Nucleotide analogue prodrug tenofovir disoproxil enhances lymphoid cell loading following oral administration in monkeys. AB - The antiviral drug tenofovir (TFV) is orally administered as the fumarate salt of its disoproxil prodrug (TFV disoproxil fumarate (TDF)). TFV is a dianion at physiological pH and, as a result, has poor lipid membrane permeability. Administration of the lipophilic and cell-permeable prodrug, TFV disoproxil, enhances the oral absorption of TFV. In order to determine whether oral administration of TDF also increases distribution to sites of viral infection, the plasma and circulating lymphoid cell pharmacokinetics of TFV and its phosphorylated metabolites were assessed following a single oral TDF or subcutaneous TFV administration at doses yielding equivalent plasma exposures to TFV in macaques. Despite TFV disoproxil's lack of plasma stability and undetectable levels in the first plasma samples taken, oral administration of TDF resulted in 7.9-fold higher peripheral blood mononuclear cell exposures to the active metabolite, TFV-diphosphate. The apparent plasma terminal half-life (t(1/2)) of TFV was also longer following oral TDF relative to subcutaneous TFV administration (median t(1/2) of 15.3 and 3.9 h, respectively), suggesting broader distribution to cells and tissues outside of the central plasma compartment. In conclusion, the disoproxil pro-moiety enhances not only the oral absorption of TFV but also tissue and lymphoid cell loading. These results illustrate that administration of even a fleeting prodrug can increase target tissue loading and give valuable insight for future prodrug development. PMID- 19545171 TI - Bases in 16S rRNA important for subunit association, tRNA binding, and translocation. AB - Ribosomes are the cellular machinery responsible for protein synthesis. A well orchestrated step in the elongation cycle of protein synthesis is the precise translocation of the tRNA-mRNA complex within the ribosome. Here we report the application of a new in vitro modification-interference method for the identification of bases in 16S rRNA that are essential for translocation. Our results suggest that conserved bases U56, U723, A1306, A1319, and A1468 in 16S rRNA are important for translocation. These five bases were deleted or mutated so their role in translation could be studied. Depending on the type of mutation, we observed inhibition of growth rate, subunit association, tRNA binding, and/or translocation. Interestingly, deletion of U56 or A1319 or mutation of A1319 to C showed a lethal phenotype and were defective in protein synthesis in vitro. Further analysis showed that deletion of U56 or A1319 caused defects in 30S subunit assembly, subunit association, and tRNA binding. In contrast, the A1319C mutation showed no defects in subunit association; however, the extent of tRNA binding and translocation was significantly reduced. These results show that conserved bases located as far as 100 A from the tRNA binding sites can be important for translation. PMID- 19545172 TI - SWI/SNF-independent nuclease hypersensitivity and an increased level of histone acetylation at the P1 promoter accompany active transcription of the bone master gene Runx2. AB - The Runx2 transcription factor is essential for skeletal development as it regulates expression of several key bone-related genes. Multiple lines of evidence indicate that expression of the Runx2/p57 isoform in osteoblasts is controlled by the distal P1 promoter. Alterations of chromatin structure are often associated with transcription and can be mediated by members of the SWI/SNF family of chromatin remodeling complexes, or by transcriptional coactivators that possess enzymatic activities that covalently modify structural components of the chromatin. Here, we report that a specific chromatin remodeling process at the proximal region (residues -400 to 35) of the Runx2 gene P1 promoter accompanies transcriptional activity in osteoblasts. This altered chromatin organization is reflected by the presence of two DNase I hypersensitive sites that span key regulatory elements for Runx2/p57 transcription. Chromatin remodeling and transcription of the Runx2 gene are associated with elevated levels of histone acetylation at the P1 promoter region and binding of active RNA polymerase II and are independent of the activity of the SWI/SNF chromatin remodeling complex. Changes in chromatin organization at the P1 promoter are stimulated during differentiation of C2C12 mesenchymal cells to the osteoblastic lineage by treatment with BMP2. Together, our results support a model in which changes in chromatin organization occur at very early stages of mesenchymal differentiation to facilitate subsequent expression of the Runx2/p57 isoform in osteoblastic cells. PMID- 19545173 TI - Structure and concentration changes affect characterization of UGT isoform specific metabolism of isoflavones. AB - We characterized the isoform specific glucuronidation of six isoflavones, genistein, daidzein, glycitein, formononetin, biochanin A and prunetin, using 12 expressed human UGTs and human intestinal and liver microsomes. The results indicated that these isoflavones are metabolized most rapidly at three different concentrations by one of these four UGT isoforms: UGT1A1, UGT1A8, UGT1A9 and UGT1A10. Furthermore, glycitein was usually metabolized the fastest whereas prunetin the slowest. Using the rates of metabolism by 12 UGT isoforms as a means to establish the metabolic "fingerprint", we found that each isoflavone had distinctive concentration-dependent patterns. Determination of kinetic parameters of glucuronidation using genistein and prunetin indicated that the distinct concentration-dependent metabolic patterns were the result of differences in K(m) and V(max) values. We then measured how well metabolic "fingerprinting" predicted metabolism of these isoflavones by human intestinal and liver microsomes. We found that the prediction was rather successful for five isoflavones in the liver microsomes, but not successful in the intestinal microsomes. We propose that a newly discovered UGT3A1 isoform capable of metabolizing phenols and estrogens might be responsible for the metabolism of isoflavones such as formononetin in humans. In conclusion, the first systematic study of metabolic "fingerprinting" of six common isoflavones showed that each isoflavone has UGT isoform-specific metabolic patterns that are concentration-dependent and predictive of metabolism of the isoflavones in liver microsomes. PMID- 19545175 TI - Quantitative expression profile of hepatobiliary transporters in sandwich cultured rat and human hepatocytes. AB - As sandwich cultured (SC) hepatocytes can repolarize to form bile canalicular networks, allowing active excretion of compounds in a vectorial manner, the model has been widely used for assessing the transporter related complexity of ADME/tox issues. A lack of quantitative information on transporter expression during cell culture has made in vitro to in vivo extrapolation of hepatobiliary transport difficult. In the present study, using our newly developed LC-MS/MS absolute quantitative methods, we determined the quantitative expression profile of three biliary transporters in SC rat and human hepatocytes. A significant shift of hepatobiliary transporter proteins was observed both in human and rat sandwich cultures. A decrease of BSEP/Bsep protein and an increase of BCRP/Bcrp protein were detected in both rat and human hepatocytes over time in culture. Interestingly, Mrp2 in rat hepatocytes was significantly diminished, while MRP2 constantly increased in human hepatocytes during the cell culture. Consequently, the interspecies difference between rat and human in absolute amount of MRP2/Mrp2 was minimized over time in culture. Following the sandwich culture, the species difference of hepatobiliary transporter protein between human and rat at day 5 post SC was diminished (MRP2/Mrp2), identical (BSEP/Bsep) or reversed (BCRP/Bcrp), compared to the in vivo situation. In addition, the absolute protein amount of BCRP/Bcrp or MRP2/Mrp2 was proportionally correlated with the intrinsic biliary clearance estimated in various lots of SC rat and human hepatocytes. The results revealed that absolute protein amount is a key determinant for hepatobiliary clearance and could provide fundamental support on extrapolation of biliary secretion from in vitro to in vivo. PMID- 19545174 TI - Rapid photothermal intracellular drug delivery using multiwalled carbon nanotubes. AB - Carbon nanotubes are unique materials that absorb infrared (IR) radiation, especially between 700 and 1100 nm, where body tissues are most transparent. Absorbed IR promotes molecular oscillation leading to efficient heating of the surrounding environment. A method to enhance drug localization for peritoneal malignancies is perfusion of warm (40-42 degrees C) chemotherapeutic agents in the abdomen. However, all tissues in the peritoneal cavity are subjected to enhanced drug delivery due to increased cell membrane permeability at hyperthermic temperatures. Here we show that rapid heating (within ten seconds) of colorectal cancer cells to 42 degrees C, using infrared stimulation of nanotubes as a heat source, in the presence of the drugs oxaliplatin or mitomycin C, is as effective as two hours of radiative heating at 42 degrees C for the treatment of peritoneal dissemination of colorectal cancer. We demonstrate increased cell membrane permeability due to hyperthermia from multiwalled carbon nanotubes in close proximity to cell membranes and that the amount of drug internalized by colorectal cancer cells heated quickly using carbon nanotubes equals levels achieved during routine application of hyperthermia at 42 degrees C. This approach has the potential to be used as a rapid bench to bedside clinical therapeutic agent with significant impact for localizing chemotherapy agents during the surgical management of peritoneal dissemination of colorectal cancer. PMID- 19545176 TI - Catalytic mechanism of SHCHC synthase in the menaquinone biosynthesis of Escherichia coli: identification and mutational analysis of the active site residues. AB - (1R,6R)-2-Succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate (SHCHC) synthase (MenH) is an alpha/beta fold enzyme containing a catalytically essential serine histidine-aspartate triad typical of serine proteases but catalyzes a pyruvate elimination reaction initiated by alpha-proton abstraction in the menaquinone biosynthetic pathway of Escherichia coli. In this study, we identify the active site residues in the synthase through sequence analysis and structural modeling and study their mechanistic roles in MenH catalysis. Steady-state kinetic characterization of site-directed mutants of the active site residues shows that three conserved arginine residues (Arg-90, Arg-124, and Arg-168) likely form ionic salt bridges with three carboxylate groups of the substrate in the Michaelis complex and that the side-chain polar groups of the conserved tyrosine (Tyr-85) and tryptophan (Trp-147) residues likely donate hydrogen bonds to form an "oxyanion hole". In addition, the pH dependence of the MenH kinetic properties reveals a catalytic base with a pK(a) highly dependent on the hydroxyl group of the triad serine residue in the enzymatic reaction. Moreover, proton inventory experiments demonstrate that the SHCHC synthase adopts one-proton catalysis like many serine proteases. These results allow the proposal of a mechanism in which the histidine residue of the MenH triad serves as a general base catalyst to deprotonate the triad seryl hydroxyl group in the alpha-proton abstraction from the substrate. As such, the MenH triad performs a simple and fundamental proton transfer reaction occurring repeatedly in the reactions catalyzed by serine proteases and alpha/beta fold hydrolases, suggesting a common evolutionary origin for all serine-histidine-aspartate triads serving different catalytic functions. PMID- 19545177 TI - Tribolium castaneum larval gut transcriptome and proteome: A resource for the study of the coleopteran gut. AB - Tribolium castaneum is an important agricultural pest and an advanced genetic model for coleopteran insects. We have taken advantage of the recently acquired T. castaneum genome to identify T. castaneum genes and proteins in one of the more critical environmental interfaces of the insect, the larval alimentary tract. Genetic transcripts isolated from the T. castaneum larval gut were labeled and hybridized to a custom array containing oligonucleotides from predicted genes in the T. castaneum genome. Through a ranking procedure based on relative labeling intensity, we found that approximately 17.6% of the genes represented in the array were predicted to be highly expressed in gut tissue. Several genes were selected to compare relative expression levels in larval gut, head, or carcass tissues using quantitative real-time PCR, and expression levels were, with few exceptions, consistent with the gut rankings. In parallel with the microarrays, proteins extracted from the T. castaneum larval gut were subjected to proteomic analysis. Two-dimensional electrophoretic analysis combined with MALDI-TOF resulted in the identification of 37 of 88 selected protein samples. As an alternative strategy, one-dimensional electrophoretic separation of T. castaneum larval gut proteins followed by two-dimensional nano-HPLC and ESI-MS/MS resulted in the identification of 98 proteins. A comparison of the proteomic studies indicated that 16 proteins were commonly identified in both, whereas 80 proteins from the proteomic analyses corresponded to genes with gut rankings indicative of high expression in the microarray analysis. These data serve as a resource of T. castaneum transcripts and proteins in the larval gut and provide the basis for comparative transcriptomic and proteomic studies related to the gut of coleopteran insects. PMID- 19545179 TI - Doing more with less: effective research and peer review. PMID- 19545181 TI - Perspectives on risk assessment for orphaned and abandoned mines. PMID- 19545180 TI - Can volcanic ash poison water supplies? PMID- 19545183 TI - Context, toxicity reference values, and risk. PMID- 19545184 TI - Silver nanoparticle toxicity and biocides: need for chemical speciation. PMID- 19545185 TI - The Nexus between ecological risk assessment and natural resource damage assessment under CERCLA: introduction to a Society of Environmental Toxicology and ChemistryTechnical Workshop. AB - A SETAC Technical Workshop titled "The Nexus Between Ecological Risk Assessment and Natural Resource Damage Assessment Under CERCLA: Understanding and Improving the Common Scientific Underpinnings," was held 18-22 August 2008 in Gregson, Montana, USA, to examine the linkage, nexus, and overlap between ecological risk assessment (ERA) and natural resource damage assessment (NRDA) under the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA). Experts from a broad range of relevant scientific, legal, and policy disciplines convened to 1) ascertain the potential for improved scientific harmonization of the processes of ERA and NRDA; 2) identify where statutory, regulatory, or scientific constraints might exist that would constrain or preclude the harmonization of the 2 processes; 3) determine approaches that might overcome these constraints; and 4) recommend research or potential changes in regulatory policies that might serve to improve both processes. This is the introduction to a series of 3 papers that describe the findings and conclusions of this workshop. Although unanimity was not achieved on all technical, legal, or policy questions posed to the participants, some consensus areas did arise. First, there appear to be few if any legal constraints to using the environmental data collected for ERA or NRDA for both processes. Second, although it is important to recognize and preserve the distinctions between ERA and NRDA, opportunities for data sharing exist, particularly for the characterization of environmental exposures and derivation of ecotoxicological information. Thus, effective coordination is not precluded by the underlying science. Where a cooperative, interactive process is involved among the response agencies, the natural resource trustees, and the responsible party(s), technical, legal or regulatory constraints can be minimized. Finally, one approach that might enhance the potential applicability of data collected for the ERA is to consider ecosystem services in the development of assessment endpoints. These points are explained in greater detail in the series of papers published herein. PMID- 19545186 TI - Ecological risk assessment and natural resource damage assessment: synthesis of assessment procedures. AB - The Society of Environmental Toxicology and Chemistry (SETAC) convened an invited workshop (August 2008) to address coordination between ecological risk assessment (ERA) and natural resource damage assessment (NRDA). Although ERA and NRDA activities are performed under a number of statutory and regulatory authorities, the primary focus of the workshop was on ERA and NRDA as currently practiced in the United States under the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA). This paper presents the findings and conclusions of the Synthesis Work Group, 1 of 3 work groups convened at the workshop. The Synthesis Work Group concluded that the different programmatic objectives and legal requirements of the 2 processes preclude development of a single, integrated ERA/NRDA process. However, although institutional and programmatic impediments exist to integration of the 2 processes, parties are capitalizing on opportunities to coordinate technical and scientific elements of the assessments at a number of locations. Although it is important to recognize and preserve the distinctions between ERA and NRDA, opportunities for data sharing exist, particularly for the characterization of environmental exposures and derivation of ecotoxicological information. Thus, effective coordination is not precluded by the underlying science. Rather, willing participants, accommodating schedules, and recognition of potential efficiencies associated with shared data collection can lead to enhanced coordination and consistency between ERA and NRDA. PMID- 19545187 TI - Synlophe structure for species of Longistrongylus (Nematoda: Trichostrongyloidea), abomasal parasites among ungulates from sub-Saharan Africa, with comparisons to the global ostertagiine fauna. AB - The synlophe, or system of longitudinal cuticular ridges characteristic of some trichostrongyloid nematodes, is examined in detail for 6 of 8 species in Longistrongylus (Ostertagiinae) that occur in ungulates across sub-Saharan Africa. Among the species of Longistrongylus examined, 5 are characterized by a tapering pattern laterally in the cervical zone (anterior to the esophageal intestinal junction), which is largely consistent among multiple male and female specimens; in contrast, for Longistrongylus meyeri the lateral pattern is parallel. The synlophe is bilaterally symmetrical, with ridges extending from the base of the cephalic expansion to near the caudal extremity in males and females. Ridges are acutely pointed, with perpendicular orientation and absence of gradient as viewed in transverse section. Species-specific patterns in conjunction with the numbers of ridges may serve to augment an array of diagnostic characters for species of Longistrongylus and contribute to increasingly accurate identification of female specimens. Among 5 of 6 species examined in the current study, the numbers of ridges in males was equal to or exceeded that observed in females, a pattern seen only in Africanastrongylus among the 15 genera of the Ostertagiinae. The differential numbers of ridges in males and females may represent another character among the suite of attributes that in part diagnose the genus Longistrongylus . PMID- 19545188 TI - First intermediate host of the digenean trematode Proctoeces lintoni (Fellodistomidae) in Chile. AB - The digenetic trematode, Proctoeces lintoni, is a parasite of the rocky intertidal ecosystems of the Chilean Pacific coastline. Although this species is relatively well known, the first intermediate host has not yet been described. In the present study, we used experimental protocols and field studies to identify the host that harbors the sporocysts and cercariae stages of the parasite. The first intermediate host was shown to be the dominant mussel of the mid-littoral zone, Perumytilus purpuratus. PMID- 19545189 TI - Translating ecological risk to ecosystem service loss. AB - Hazardous site management in the United States includes remediation of contaminated environmental media and restoration of injured natural resources. Site remediation decisions are informed by ecological risk assessment (ERA), whereas restoration and compensation decisions are informed by the natural resource damage assessment (NRDA) process. Despite similarities in many of their data needs and the advantages of more closely linking their analyses, ERA and NRDA have been conducted largely independently of one another. This is the 4th in a series of papers reporting the results of a recent workshop that explored how ERA and NRDA data needs and assessment processes could be more closely linked. Our objective is to evaluate the technical underpinnings of recentmethods used to translate natural resource injuries into ecological service losses and to propose ways to enhance the usefulness of data obtained in ERAs to the NRDA process. Three aspects are addressed: 1) improving the linkage among ERA assessment endpoints and ecological services evaluated in the NRDA process, 2) enhancing ERA data collection and interpretation approaches to improve translation of ERA measurements in damage assessments, and 3) highlighting methods that can be used to aggregate service losses across contaminants and across natural resources. We propose that ERA and NRDA both would benefit by focusing ecological assessment endpoints on the ecosystem services that correspond most directly to restoration and damage compensation decisions, and we encourage development of generic ecosystem service assessment endpoints for application in hazardous site investigations. To facilitate their use in NRDA, ERA measurements should focus on natural resource species that affect the flow of ecosystem services most directly, should encompass levels of biological organization above organisms, and should be made with the use of experimental designs that support description of responses to contaminants as continuous (as opposed to discrete) variables. Application of a data quality objective process, involving input from ERA and NRDA practitioners and site decision makers alike, can facilitate identification of data collection and analysis approaches that will benefit both assessment processes. Because of their demonstrated relationships to a number of important ecosystem services, we recommend that measures of biodiversity be targeted as key measurement endpoints in ERA to support the translation between risk and service losses. Building from case studies of recent successes, suggestions are offered for aggregating service losses at sites involving combinations of chemicals and multiple natural resource groups. Recognizing that ERA and NRDA are conducted for different purposes, we conclude that their values to environmental decision making can be enhanced by more closely linking their data collection and analysis activities. PMID- 19545190 TI - Effective coordination and cooperation between ecological risk assessments and natural resource damage assessments: a new synthesis. AB - Although ecological risk assessments (ERAs) and natural resource damage assessments (NRDAs) are performed under different statutory and regulatory authorities, primarily the Comprehensive Environmental Response, Compensation and Liability Act (CERCLA), as currently practiced, the activities typically overlap. ERAs performed as part of the response process (typically by the US Environmental Protection Agency [USEPA]) should be closely coordinated with the natural resource trustees' (trustees') NRDAs. Trustees should actively participate in the early stages of the remedial investigation (RI) and work with USEPA, including the potentially responsible parties (PRPs), when appropriate, to coordinate NRDA data needs with those of the RI. Close coordination can present opportunities to avoid inefficiencies, such as unnecessary resampling or duplicate data gathering, and provide the opportunity to fulfill both process requirements with a few well designed investigations. Early identification of opportunities for practical combined assessment can save money and time as the restoration process proceeds and facilitate a cooperative resolution of the entire site's CERCLA liability. The Society of Environmental Toxicology and Chemistry (SETAC) convened an invited workshop (August 2008) to address coordination between ERA and NRDA efforts. This paper presents the findings and conclusions of the Framework Work Group, which considered technical issues common to each process, while mindful of the current legal and policy landscape, and developed recommendations for future practice. PMID- 19545191 TI - A new genus and species of mermithid (Nematoda) emerging from chironomid imagos eclosing from the Mississippi River headwaters area in northern Minnesota. AB - Fibromermis roarki n. gen., n. sp. (Nematoda: Mermithidae) is described from the hemocoel of chironomid imagos of Ablabesmyia monilis (L.) eclosing from the headwaters of the Mississippi River at Gulsvig Landing in northern Minnesota in the summer of 2007. The new genus is described, distinguished from the most closely related tetrapapillate aquatic mermithid genus, and the new species described. Intensity and sex specificity of infection are recorded from 28 host adult chironomids of a single cohort. The new species represents the fourth tetrapapillate mermithid species described from the area and the third tetrapapillate genus. PMID- 19545192 TI - Internet tool box for rural GPs to access mental health services information. AB - INTRODUCTION: Rural GPs play a significant role in the mental health care of their patients. It is therefore crucial that they have access to reputable support and advice that enhances their existing knowledge. This article outlines a recent project initiated by the Australian rural Ballarat and District Division of General Practice (BDDGP) to develop and implement an online resource to facilitate local implementation and delivery of the 'Better Access to Mental Health Care' (BAMHC) program. This 12 month project was initiated in response to a request from local GPs for additional information about and support in using the BAMHC program. The project is the culmination of significant collaboration among key stakeholders that includes local GPs, GP advisors from BDDGP, BDDGP staff, and two University of Ballarat research centres (the Centre for Health Research and Practice, and the Centre for Electronic Commerce and Communication). This article documents the key stages involved in the project from initiation to implementation, and reports on the use of this resource 12 months after its launch. METHOD: The BDDGP represents 107 GPs and six GP registrars and covers a large rural/semi-rural area of 7300 km2 and a catchment population of more than 120,000. The format and design of the project entailed four distinct but interrelated stages of development: (1) developing the program specifications and localising it to the BDDGP catchment; (2) constructing a decision-making support tool with 7 sequential steps comprising key questions and links to detailed answers; (3) developing and populating an online service directory of local allied health professionals; and (4) constructing the website for easy access and navigation for GPs and other service providers. RESULTS: The website was publicly launched in December 2007 and is hosted by BDDGP. Since then it has received strong support. In the 12 months since its launch the website received regular and continuous visits (2847 visits/11,500 pages accessed). In addition, anecdotal evidence and other feedback (positive comments; requested changes to entries in the service directory from allied mental health professionals) indicate that the website is being recognised as an important resource of and hub for local information relating to the BAMHC program for GPs and allied health professionals. CONCLUSIONS: Integral to the website's success and sustainability is the close and continued monitoring and updating of the information provided. A formal, longitudinal evaluation 18 months to 2 years after the website's launch is recommended to provide a more rigorous assessment of the tool, and examine possible improvements. While the project does not address the problem of the supply of allied mental health providers in rural areas, it does provide assistance with responsive service system expansion and the provision of a localized tool for accessing appropriate information about mental health services. PMID- 19545193 TI - Positive hyaluronan/PEI/DNA complexes as a target-specific intracellular delivery to malignant breast cancer. AB - In the present study, The PEI/DNA (PD) complexes was first prepared with positive surface charge under a suitable N/P ratio of 10. The redundant positive charge was partially and excessively shielded by a polysaccharide, hyaluronic acid (HA), in aqueous solution. The HA/PEI/DNA ternary complexes were characterized by assessing the zeta potential and size, then transferred to MDA-MB-435, MDA-MB 231, and MCF-7 cell lines with different amounts of HA-specific CD44 receptors on the surface. Consequently, The transfection efficiency of all the prepared complexes show a little increased to MCF-7 (low CD44 level) while a large increased to MDA-MB-231 and MDA-MB-435 cells (high CD44 level) with adding HA. Also, when HA:PEI charge ratio was 7.5%, the ternary complexes show the highest transfection efficiency. The prepared ternary complexes exhibited increased 2-13 fold fluorescence intensity and lower cell toxicity compared to the PD (N/P, 10). These results indicated that the positive HA/PEI/DNA ternary complexes (HA:PEI charge ratio, 7.5%) can target malignant breast cancer cells with high CD44 level and might be a promising candidate vector for gene therapy. PMID- 19545196 TI - Risk assessment of chemical carcinogens and thresholds. AB - The controversial arguments about the existence of "thresholds" for carcinogens are discussed and some conclusions are drawn: (1) The meaning of "threshold" has changed considerably during the last decades. Initially, the discussion focused on the genotoxic properties of chemicals. In dose-response studies the endpoint was tumor incidence. Later, DNA adducts represented the biologically active target dose and whether saturation of metabolic activation could lead to non linear relationships was tested as a hypothesis. (2) In a next step, the implications of the initiation-promotion model were studied. Carcinogens with tumor-initiating properties showed linear dose-response relationships at low doses without a definable threshold, whereas those with tumor-promoting properties showed non-linear characteristics compatible with the existence of a threshold. However, the results are difficult to transfer to the human situation, and many critical endpoints are subject to other risk factors so that a meaningful value cannot be given. (3) Eventually, it turned out that most carcinogens exhibit genotoxic as well as non-genotoxic properties, and toxicity may be equally important as genotoxicity. (4) In view of the discussion for more than 60 years about the existence of thresholds for carcinogens, it is suggested that the threshold approach not be used to establish acceptable risk limits. (5) Instead of calculating an acceptable risk from cancer risk data, the recommended method is to assess the incremental contribution of exposure to the background of avoidable and unavoidable exposures by using biomonitoring data from human individuals. Such data could help in risk management, in order to reach acceptable limits of exposures on the basis of the "as low as reasonably achievable" or "ALARA" principle. PMID- 19545194 TI - Preparation of self-assembled microspheres and their potential for drug delivery. AB - Dextran solutions intended for use as plasma extenders have been observed to form insoluble precipitates. Earlier studies of precipitation have shown that in solutions of 50% and 60% w/w of dextran molecular mass 6000 g mol(-1) beaded precipitates are formed over a two-week period. This study considers dextran precipitation over a wider molecular mass range and the kinetics, of formation, morphology and potential utility of these precipitates is investigated. Results show precipitation occurs over the dextran molecular mass range 6000-17,000 g mol(-1), with lower molecular mass material showing more rapid precipitation. As bead formation is accompanied by an increase in turbidity, formation kinetics were quantified spectrophotometrically confirming that precipitation rates were inversely proportional to molecular mass. The utility of these precipitates for drug delivery applications was assessed using bovine serum albumin as a protein drug analogue. The results showed that the inclusion of protein did not prevent bead formation and that entrapped protein was subsequently released from dextran beads in a time dependant manner. This suggests that dextran beads of this type may find application in the drug delivery area, as they combine the advantages of mild entrapment conditions with the use of an unmodified clinically approved polymer. PMID- 19545197 TI - Derivation of inhalation toxicity reference values for propylene oxide using mode of action analysis: example of a threshold carcinogen. AB - Propylene oxide (PO) is an important industrial chemical used primarily in the synthesis of other compounds. Inhalation carcinogenesis studies in rodents, with no-observed-adverse-effect levels (NOAELs) of 100 and 200 ppm, have revealed that chronic, high exposure to PO can induce tumors at the site of contact. Despite these characteristics, there is no evidence that typical environmental or occupational exposures to PO constitute a health risk for humans. The nongenotoxic effects of PO (glutathione depletion and cell proliferation) that augment its DNA-reactive and non-DNA-reactive genotoxicity are expected to be similar in humans and rodents. Available evidence on mode-of-action suggests that cancer induction by PO at the site of contact in rodents is characterized by a practical threshold. Human toxicity reference values for potential carcinogenic effects of PO were derived based on nasal tumors identified in rodent studies and specified uncertainty factors. The 95% lower confidence limit on the dose producing a 10% increase in additional tumor risk (LED10) was calculated using the rat and mouse data sets. The human reference values derived from the rat and mouse LED10 values were 0.7 and 0.5 ppm PO, respectively. A similar noncancer reference value, 0.4 ppm, was derived on the basis of non-neoplastic nasal effects in rats. PMID- 19545198 TI - The inhalation toxicology of p-aramid fibrils. AB - The pandemic of lung disease caused by asbestos has cast suspicion on any industrial fibrous material that can become airborne in respirable form in workplaces, such that the respirable fibres might be inhaled. Fibre toxicology arose as a sub-specialty of particle toxicology to address the specialised nature of fibre effects and has evolved substantially in the last 25 years. It has yielded valuable information on the dosimetry, structure-activity relationships, and mechanism involved in toxicological effects of a range of fibrous materials, including asbestos, other naturally occurring fibrous materials, and synthetic vitreous fibres. A robust structure/activity paradigm has emerged from this research that highlights fibre length, thinness, and biopersistence as major factors in determining the pathogenicity of a fibre. p-Aramid is a manufactured fibre composed of synthetic polyamide (poly paraphenylene terephthalamide) manufactured on a commercial scale since 1970 by polymerisation and spinning steps. It is used as an advanced composite and in fabrics, body armour, friction materials, etc. Respirable fibrils of p-aramid can be released from the fibres during working and can become airborne. A considerable body of research has been carried out into the hazard posed by inhaled p-aramid fibrils, and this review considers this body of literature and summarises the state-of-the-science in the toxicology of p-aramid fibrils in the light of the existing overarching fibre toxicology paradigm. The peer-reviewed studies demonstrate that p-aramid fibrils can be long and thin but that the fibrils are not biopersistent. Residence in the milieu of the lungs leads to fibre shortening, allowing efficient and complete phagocytosis and effective clearance. Subsequently the p-aramid hazard is low, and this is confirmed in animal studies. The mechanism of shortening of p-aramid fibrils is not well-understood, but may involve the action of macrophages on the fibrils following phagocytosis. PMID- 19545199 TI - Induction of tunica vaginalis mesotheliomas in rats by xenobiotics. AB - To better understand the relevance of tunica vaginalis mesotheliomas (TVM) to human cancer risk, we examined the nature of TVM responses in 21 published rat cancer bioassays against the backdrop of the biology and molecular biology of mesothelium, and of spontaneous and treatment-induced TVM. Although relatively rare in all species including humans, TVM are seen most frequently in F344 male rats, as opposed to other rat strains, and are causally associated with the high background incidence of Leydig-cell tumors of the testes of these rats. Hormone imbalance brought about by perturbations of the endocrine system is proposed as a key factor leading to both spontaneous and treatment-associated TVM. Of 21 F344 rat studies with a treatment-associated TVM response, 7 were judged to have a nonsignificant to marginal response, 11 had a robust TVM response, and 3 were noninformative due to early mortality from other induced tumors. Of the 11 chemicals with robust responses, 8 were directly mutagenic in Salmonella and 3 are known to be mutagenic after metabolism. Only 2 of the 7 with nonsignificant to marginal responses were Ames test positive. TVM induction is a male F344 rat specific event, and chemicals/agents that induce only TVM in the male F344 rat from a typical two-sex rat and mouse chronic bioassay are likely irrelevant in human risk assessment. PMID- 19545200 TI - Effects of mercury on the endocrine system. PMID- 19545203 TI - Optimization of gatifloxacin liposomal hydrogel for enhanced transcorneal permeation. AB - The aim of this study was to prepare and characterize a topically effective prolonged-release ophthalmic gatifloxacin liposomal hydrogel formulation. Reverse phase evaporation was used for the preparation of liposomes consisting of phosphatidylcholine (PC) and cholesterol (CH). The effect of PC:CH molar ratio on the percentage of drug encapsulated was investigated. The effect of additives, such as stearylamine (SA) or dicetyl phosphate (DP), as positive and negative charge inducers, respectively, was studied. Morphology, mean size, encapsulation efficiency, and in vitro release of gatifloxacin from liposomes were evaluated. For hydrogel preparation, carbopol 940 was applied. In vitro transcorneal permeation through excised albino rabbit cornea was also determined. Optimal encapsulation efficiency was found at the 5:3 PC:CH molar ratio; by increasing CH content above this limit, the encapsulation efficiency decreased. Positively charged liposomes showed superior entrapment efficiency over other liposomes. Hydrogel-containing liposomes with lipid content PC, CH, and SA in a molar ratio of 5:3:1, respectively, showed best release and transcorneal permeation. These results suggest that the encapsulation of gatifloxacin into liposomes prolonged the in vitro release, depending on composition of the vesicles. In addition, the polymer hydrogel used in the preparation ensured steady, prolonged transcorneal permeation. In conclusion, gatifloxacin liposomal hydrogel is a suitable delivery system for the improvement of the ocular bioavailability of gatifloxacin. PMID- 19545204 TI - Reconstruction of an in vitro tissue-specific microenvironment to rejuvenate synovium-derived stem cells for cartilage tissue engineering. AB - Joint injury results in cartilage lesions that are characterized by a poor repair response. Adult stem cells are immensely appealing for biological joint repair, such as cartilage tissue engineering and regeneration. However, adult stem cells gradually lose their stemness once they are removed from their in vivo niche for plating in plastic flasks. We utilized a tissue-specific stem cell, synovium derived stem cell (SDSC), as a model to reconstruct an in vitro three-dimensional stem cell niche. After seeding on SDSC-derived extracellular matrix, the initially wide and flat SDSCs became thin and spindle shaped and were arranged in a three-dimensional configuration with typical stem cell phenotypes. A dramatic increase in cell number and a greatly enhanced chondrogenic capacity were observed, though surprisingly the extracellular matrix-treated SDSCs did not display concomitantly improved adipogenic or osteogenic potentials. Thus, we conclude that a tissue-specific stem cell can be used to prepare its own in vitro niche for stem cell proliferation while maintaining and enhancing its lineage specific stemness. The ability to reconstitute the in vitro stem cell niche will greatly benefit SDSC-based therapy for cartilage defects. PMID- 19545206 TI - Ecological limits and fitness consequences of cross-gradient pollen movement in Lasthenia fremontii. AB - The interaction between gene flow and environmental heterogeneity plays a key role in shaping the distribution patterns that we observe in natural populations. Although a growing body of theoretical work is exploring the effects of gene flow on the evolution of range limits and ecological specialization, explicit empirical tests of model assumptions and predictions in natural populations are almost entirely lacking. This study examines the potential for center-to-edge gene flow to occur and estimates the fitness consequences of cross-gradient gene flow in an annual plant species restricted to California vernal pool wetlands. Phenological differences and highly focused foraging patterns of pollinators reduce the potential for center-to-edge gene flow across populations within pools. Furthermore, controlled crosses simulating different patterns of gene flow across the environmental gradient reveal that center-to-edge gene flow does not reduce plant fitness at the edge but instead yields an increase in emergence rates and a trend toward overall higher fitness. PMID- 19545205 TI - Future of polio vaccines. AB - Over the past half-century, global use of highly effective vaccines against poliomyelitis brought this disease to the brink of elimination. Mounting evidence supports the argument that a high level of population immunity must be maintained after wild poliovirus circulation is stopped to preserve a polio-free status worldwide. Shifting factors in the risk-benefit-cost equation favor the creation of new poliovirus vaccines for use in the foreseeable future. Genetically stable attenuated virus strains could be developed for an improved oral poliovirus vaccine, but proving their safety and efficacy would be impractical owing to the enormous size of the clinical trials required. Novel versions of inactivated poliovirus vaccine that could be used globally should be developed. An improved inactivated poliovirus vaccine must be efficacious, inexpensive, safe to manufacture and easy to administer. Combination products containing inactivated poliovirus vaccine and other protective antigens should become part of routine childhood immunizations around the world. PMID- 19545207 TI - Severe dysglycemia with the fluoroquinolones: a class effect? AB - BACKGROUND: Although gatifloxacin is no longer available, other fluoroquinolones may significantly interfere with glucose homeostasis. The objective of the present study was to compare the risk of severe hypo- and hyperglycemia in a cohort of patients treated with gatifloxacin, levofloxacin, ciprofloxacin, or azithromycin. METHODS: This was a retrospective inception cohort study of outpatients with a new prescription for gatifloxacin, levofloxacin, ciprofloxacin, or azithromycin from 1 October 2000 through 30 September 2005 in the Veterans Affairs health care system. For patients who received one of these antibiotics, we identified outcomes of hospitalization with a primary diagnosis of hypo- or hyperglycemia. Multivariable logistic regression was used to determine the odds of hypo- and hyperglycemia with the individual fluoroquinolones versus azithromycin. RESULTS: The crude incidence rates for severe hypo- and hyperglycemia among those who received gatifloxacin, levofloxacin, ciprofloxacin, and azithromycin were 0.35 and 0.45, 0.19 and 0.18, 0.10 and 0.12, and 0.07 and 0.10 cases per 1000 patients, respectively. Among patients with diabetes, the odds ratios for hypoglycemia compared with azithromycin were 4.3 (95% confidence interval [CI], 2.7-6.6) for gatifloxacin, 2.1 (95% CI, 1.4-3.3) for levofloxacin, and 1.1 (95% CI, 0.6-2.0) for ciprofloxacin. The odds ratios for hyperglycemia were 4.5 (95% CI, 3.0-6.9) for gatifloxacin, 1.8 (95% CI, 1.2-2.7) for levofloxacin, and 1.0 (95% CI, 0.6-1.8) for ciprofloxacin. CONCLUSIONS: The odds of severe hypo- and hyperglycemia were significantly greater with gatifloxacin and levofloxacin, but not ciprofloxacin, than with azithromycin. Thus, the risk of a clinically relevant dysglycemic event appears to vary among the fluoroquinolones. PMID- 19545208 TI - Importance of the epa locus of Enterococcus faecalis OG1RF in a mouse model of ascending urinary tract infection. AB - Previously, TX5179, a disruption mutant of the enterococcal polysaccharide antigen (epa) gene cluster of Enterococcus faecalis strain OG1RF was shown to be attenuated in translocation, biofilm, mouse peritonitis and was more susceptible to polymorphonuclear leukocyte phagocytic killing. Here, wild-type E. faecalis OG1RF and TX5179 strains were tested in a mixed-infection (inoculum, approximately 1:1) mouse urinary tract infection model. Wild-type OG1RF outnumbered TX5179 in the kidneys (P < .001) and bladder (P < .001). In conclusion, the epa locus of E. faecalis OG1RF contributes to murine urinary tract infection and is the first such enterococcal polysaccharide locus shown to be important in this site. PMID- 19545210 TI - Therapeutic monoclonal antibody treatment targeting respiratory syncytial virus (RSV) G protein mediates viral clearance and reduces the pathogenesis of RSV infection in BALB/c mice. AB - Because the G protein of respiratory syncytial virus (RSV) has a CX3C chemokine motif that has been associated with the ability of RSV G protein to modulate the virus-induced host immune response, we examined whether therapeutic treatment with an anti-RSV G monoclonal antibody (mAb), 131-2G, that blocks the CX3C associated activity of RSV G protein might decrease the pulmonary inflammation associated with infection in BALB/c mice. The results show that treatment with mAb 131-2G on day 3 after RSV infection reduces both inflammation and RSV titer in the lungs. Later administration of anti-RSV G mAb (day 5 after RSV infection) effectively reduced the viral titer but had a minimal effect on pulmonary inflammation. This study suggests that an anti-RSV G mAb might be an effective antiviral, either alone or in combination with anti-RSV F protein neutralizing antibodies, for decreasing the virus-induced host response to infection and improve treatment outcome. PMID- 19545209 TI - Adult male circumcision does not reduce the risk of incident Neisseria gonorrhoeae, Chlamydia trachomatis, or Trichomonas vaginalis infection: results from a randomized, controlled trial in Kenya. AB - BACKGROUND: We examined the effect of male circumcision on the acquisition of 3 nonulcerative sexually transmitted infections (STIs). METHODS: We evaluated the incidence of STI among men aged 18-24 years enrolled in a randomized trial of circumcision to prevent human immunodeficiency virus (HIV) infection in Kisumu, Kenya. The outcome was first incident nonulcerative STI during 2 years of follow up. STIs examined were laboratory-detected Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis infection. RESULTS: There were 342 incident infections among 2655 men followed up. The incidences of infection due to N. gonorrhoeae, C. trachomatis, and T. vaginalis were 3.48, 4.55, and 1.32 cases per 100 person-years, respectively. The combined incidence of N. gonorrhoeae and C. trachomatis infection was 7.26 cases per 100 person-years (95% confidence interval, 6.49-8.13 cases per 100 person-years). The incidences of these STIs, individually or combined, did not differ by circumcision status as a time-dependent variable or a fixed variable based on assignment. Risks for incident STIs in multivariate analysis included an STI at enrollment, multiple sex partners within <30 days, and sexual intercourse during menses in the previous 6 months; condom use was protective. CONCLUSIONS: Circumcision of men in this population did not reduce their risk of acquiring these nonulcerative STIs. Improved STI control will require more-effective STI management, including partner treatment and behavioral risk reduction counseling. PMID- 19545211 TI - A dose-escalation safety and immunogenicity study of live attenuated oral rotavirus vaccine 116E in infants: a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: Rotavirus infections cause approximately 122,000 deaths among Indian children annually. METHODS: The neonatal rotavirus candidate vaccine 116E was tested in a double-blind, placebo-controlled dose-escalation trial in India. Two doses of the Vero cell-adapted vaccine were evaluated. One hundred eighty-seven infants received a vaccine dose of 1 X 10(4) focus-forming units (ffu) and 182 received a dose of 1 X 10(5) ffu in a 1:1 randomization with placebo recipients. Infants received the vaccine at 8, 12, and 16 weeks, separately from routine vaccines. RESULTS: No significant differences in clinical adverse events or laboratory toxicity were observed between vaccine and placebo recipients. There were no vaccine-related serious adverse events. A 4-fold increase in rotavirus immunoglobulin A titer was observed in 66.7% and 64.5% of infants after the first administration and in 62.1% and 89.7% of infants after 3 administrations of doses of 1 X 10(4) ffu and 1 X 10(5) ffu, respectively; the differences between these groups and placebo recipients were statistically significant. CONCLUSIONS: Three administrations of vaccine doses of 1 X 10(4) ffu and 1 X 10(5) ffu were safe. The 1 X 10(5)-ffu dose of 116E demonstrated a robust immune response after 3 administrations. These favorable results warrant further development of the vaccine candidate and provide optimism that vaccinating infants in the developing world will prevent serious sequelae of rotavirus infection. Clinical trials registration. NCT00439660 and ISRCTN57452882 . PMID- 19545213 TI - Dietary regulation of P-gp function and expression. AB - Food-drug interactions have been associated with clinically important pharmacokinetic and pharmacodynamic changes of a drug. The aim of this paper is to review the regulation of P-glycoprotein (P-gp) by dietary components and to correlate the changes in cellular P-gp function and expression with drug bioavailability. In summary, the published literature has provided extensive data supporting the modulation of drug bioavailability through P-gp regulation by components in food groups such as fruit juices, spices, herbs, cruciferous vegetables and green tea. Most of these data were, however, derived from in vitro cell models and, except for the St John's wort, the clinical significance of most reported interactions remains to be clarified. Studies on piperine and capsaicin have underscored an often poor correlation between in vivo and in vitro data, whereas experiments involving curcumin highlighted differences between acute and chronic consumption of a dietary component on P-gp function and expression in vivo. A better understanding of the pharmacokinetic and pharmacodynamic profiles of the dietary components will aid in addressing these knowledge gaps. PMID- 19545212 TI - Highly effective oral amphotericin B formulation against murine visceral leishmaniasis. AB - Visceral leishmaniasis is a deadly parasitic disease caused by obligate intramacrophage protozoans of the Leishmania genus. The World Health Organization estimates the annual death toll to be 50,000, with 500,000 new cases each year. Without treatment, visceral leishmaniasis is inevitably fatal. For the last 70 years, the first line of defense has been pentavalent antimonials; however, increased resistance has brought amphotericin B to the forefront of treatment options. Unfortunately, the difficult route of drug administration, toxicity issues, and cost prevent amphotericin B from reaching the infected population, and mortality continues to rise. Our reformulation of amphotericin B for oral administration has resulted in a highly efficacious antileishmanial treatment that significantly reduces or eradicates liver parasitemia in a murine model of visceral leishmaniasis. This formulation has overcome amphotericin B's significant physicochemical barriers to absorption and holds promise for the development of a self-administered oral therapy for the treatment of visceral leishmaniasis. PMID- 19545214 TI - Fexofenadine: biochemical, pharmacokinetic and pharmacodynamic properties and its unique role in allergic disorders. AB - BACKGROUND: Fexofenadine is one of several second-generation H(1)-antihistamines approved for the treatment of various allergic disorders; however, it shows numerous unique properties that make it an optimal choice for many patients. OBJECTIVE: To review the pharmacology, efficacy and safety of fexofenadine and the attributes differentiating it from other H(1)-antihistamines. METHODS: We performed a literature search in PubMed/MEDLINE (1966 - March 2009) using the keywords fexofenadine, antihistamine, allergic rhinitis and chronic urticaria. We also reviewed data provided by the manufacturer in addition to reports from various governmental agencies. RESULTS/CONCLUSIONS: Fexofenadine is devoid of sedative and anticholinergic effects and may offer equivalent or greater efficacy in treating allergic disorders compared with other currently available second generation H(1)-antihistamines. In addition, fexofenadine may offer cost savings over other selected H(1)-antihistamines owing to its recent availability in generic form in the US. PMID- 19545215 TI - Tacrolimus in hematopoietic stem cell transplantation. AB - BACKGROUND: Graft-versus-host disease (GVHD) associated with allogeneic peripheral blood or marrow transplantation is a serious life-threatening complication. Most of the available literature support the use of tacrolimus as a prophylactic agent for acute GVHD. OBJECTIVE: To review the pharmacology, efficacy and safety of tacrolimus and recommend its place in therapy for the prophylaxis of GVHD. METHODS: We conducted a literature search using PubMed/Medline (January 1996 - December 2008) using the keywords tacrolimus, graft-versus-host disease and hematopoietic stem cell transplantation. Data provided by the manufacturer and the FDA were also reviewed. CONCLUSION: Tacrolimus is effective in the prevention of acute GVHD. Trials comparing tacrolimus with cyclosporine using tacrolimus levels in the 7 - 10 ng/ml range and with larger numbers of patients may be necessary to better understand the impact of these drugs on survival. PMID- 19545216 TI - Cost-effectiveness of insulin detemir compared with neutral protamine Hagedorn insulin in patients with type 1 diabetes using a basal-bolus regimen in five European countries. AB - OBJECTIVES: The aim of this analysis was to evaluate the long-term clinical and economic outcomes associated with insulin detemir and neutral protamine Hagedorn (NPH) insulin in combination with mealtime insulin aspart in patients with type 1 diabetes in Belgian, French, German, Italian and Spanish settings. METHODS: The published and validated IMS CORE Diabetes Model was used to make long-term projections of life expectancy, quality-adjusted life expectancy and direct medical costs. The analysis was based on patient characteristics and treatment effects from a 2-year randomised controlled trial. Events were projected for a time horizon of 50 years. Potential uncertainty using a modelling approach was addressed. RESULTS: Basal-bolus therapy with insulin detemir was projected to improve quality-adjusted life expectancy by 0.45 years versus NPH in the German setting, with similar improvements in the other countries. Insulin detemir was associated with cost savings in Belgium, Germany and Spain. In France and Italy, lifetime costs were slightly higher in the detemir arm, leading to incremental cost-effectiveness ratios of 519 euro per QALY gained and 3,256 euro per QALY gained, respectively. CONCLUSIONS: Compared to NPH, insulin detemir is likely to be a dominant treatment strategy in Belgium, Germany and Spain and highly cost effective in France and Italy in patients with type 1 diabetes. PMID- 19545217 TI - Non-viral vectors in cystic fibrosis gene therapy: recent developments and future prospects. AB - Gene therapy has been proposed for a wide range of human diseases but few have received the level of attention over such a prolonged period as cystic fibrosis (CF) with over 20 clinical studies undertaken. Following a 10-year interval, clinical trials of an aerosolisable non-viral gene transfer agent have recently been initiated by researchers in the United Kingdom. Here we review the rationale and requirements for effective gene therapy for CF lung disease. The previous non viral gene therapy trials are discussed and the prospects for the current leading non-viral formulations for CF gene therapy are considered. Factors affecting the selection and design of the plasmid DNA molecule, likely to be of central importance to clinical efficacy, are reviewed and we describe the potential merits of the formulation that has been selected for the forthcoming UK trials. PMID- 19545218 TI - Therapeutic implications of Cancer Initiating Cells. AB - BACKGROUND: Until few years ago, all neoplastic cells within a tumour were suggested to have tumorigenic capacity, but recent evidences hint to the possibility that such feature is confined to a subset of Cancer Initiating Cells (CICs), also called Cancer Stem Cells (CSCs). These cells are the reservoir of the heterogeneous populations of differentiated cancer cells constituting the tumour bulk. Mechanisms shared with somatic stem cells, such as quiescence, self renewal ability, asymmetric division and multidrug resistance, allow to these cells to drive tumour growth and to evade conventional therapy. OBJECTIVE: Here, we give a brief overview on the origin of CICs, the mechanisms involved in chemoresistance and therapeutic implications. CONCLUSION: Current cancer treatments, based on the assumption that tumour cell population responds homogeneously, have been developed to eradicate proliferating cells. The new model of tumorigenesis entails significant therapeutic implications, in fact if a small fraction of CICs survives conventional therapy it may lead to recurrence after month or years of apparent remission. Selective targeting of CICs could eliminate the tumour from the root, overcoming the emergence of clones capable of evading traditional therapy and increasing overall disease free survival. PMID- 19545220 TI - Effect of lecithin and starch on alginate-encapsulated probiotic bacteria. AB - The effect of lecithin and starch on viability of alginate encapsulated probiotics was determined at different temperatures. Probiotic organisms (1% v/v>10Log CFU ml(-1)) were encapsulated using alginate (2% w/v), gelatinized starches (2% w/v) and lecithin (0-4% w/v) and stored in sealed containers at 4, 23 and 37 degrees C (to simulate shelf storage conditions). Incorporation of lecithin improved the entrapment efficiency (p < 0.05) and the viability of encapsulated bacteria (p = 0.02). Encapsulated Lactobacillus, Bifidobacterium species and Lactococcus lactis in lecithin containing freeze-dried beads had good survival stability (above 6Log CFU ml(-1)) at 23 degrees C for 12 weeks. The bacteria in the beads showed 6Log survival by the end of 2 weeks at 37 degrees C. Encapsulated L. casei in the alginate beads containing lecithin were also more stable in the yoghurt than the beads without lecithin. SEM analysis of the beads showed an irregular surface for the beads without lecithin. PMID- 19545219 TI - Encapsulation of liquid cores by layer-by-layer adsorption of polyelectrolytes. AB - The aim of this work was to develop the method of preparation of loaded, submicron nanocapsules based on the liquid core encapsulation by polyelectrolyte (PE) multilayer adsorption. The procedure of PE adsorption on the emulsion droplets requires a specific selection of surfactants, which have good properties as emulsifiers and provide a stable surface charge for sequential adsorption of PE without losing stability of emulsion. Using AOT as emulsifier this study obtained droplets, stabilized by AOT/PDADMAC surface complexes. These positively charged liquid cores were then modified by sequential adsorption of polyelectrolytes to obtain nanocapsules with the average size of 200 nm, with various combinations of polyelectrolytes (PDADMAC, CHIT, PAH, PSS, ALG). This study demonstrated the formation of consecutive layers of PE shells by measuring zeta potential of capsules after adsorption of each layer. It visualized the cores by dissolving fluorescent dye Coumarine6 in oil phase and multilayer shells by using FITC labelled polycation. PMID- 19545221 TI - Improved delivery of angiogenesis inhibitors from PLGA:poloxamer blend micro- and nanoparticles. AB - Clinical studies have demonstrated the efficacy of new strategies in cancer therapy, such as chemotherapy and radiotherapy, associated to the administration of tumour vascularization inhibitors. A critical limitation for the clinical application of angiogenesis inhibitors relies in their instability in biological environment and high-dose requirements. This work has attempted to overcome this limitation by designing an adequate delivery vehicle consisting of PLGA:poloxamer blend micro- and nanoparticles. The potential of this delivery system was investigated for a new synthetic angiogenesis inhibitor named polyaminoacid JS 2892b. PLGA:poloxamer (ratio 10 : 1) blend microparticles were prepared by the oil-in-oil emulsion technique, while PLGA:poloxamer (ratio 1 : 1) blend nanoparticles were obtained by a modified solvent diffusion technique. The results showed that, by adjusting the formulation conditions, it was possible to efficiently encapsulate the polyaminoacid JS-2892b within PLGA:poloxamer micro- (particle size of 20 microm and encapsulation efficiency higher than 90%) and nanoparticles (particle size of less than 280 nm and encapsulation efficiency of 52%). In addition, the delivery of the polyaminoacid JS-2892b from the particles could be controlled, without altering its stability, for extended periods of time (from a few days to over a month). The release of the encapsulated compound was significantly affected by the particle size and the way the drug is dispersed into the polymeric matrix. Therefore, this study provides information about the formulation conditions and potential of biodegradable particles for the controlled release of polyaminoacid JS-2892b. PMID- 19545223 TI - In vitro and in vivo evaluations of PLGA microsphere vaccine formulations containing pDNA coexpressing hepatitis B surface antigen and Interleukin-2. AB - PURPOSE: DNA-based vaccines encoding viral antigens have been shown to elicit immune responses in animal models. In this study, a plasmid DNA (pDNA) coexpressing the middle envelope protein of hepatitis B virus (HBV) and Interleukin-2 (IL-2) was incorporated into Poly (D,L-lactic-co-glycolic acid) (PLGA) microspheres and three different formulations were investigated for their potential as a vaccine delivery system. METHODS: Emulsion solvent evaporation methods of water-in-oil-in-water (w/o/w) and oil-in-water (o/w) were used to generate three different formulations in which PLGA microspheres contained pDNA either encapsulated within or adsorbed onto the microspheres. RESULTS: In vaccine formulation A2, prepared using the (w/o/w) method, pDNA was encapsulated within the microspheres. The other two formulations (B2 and B2a) were prepared using the (o/w) method and B2 contained pDNAs encapsulated within the microspheres while B2a contained pDNAs adsorbed onto the microspheres. pDNA loading efficiencies of A2, B2 and B2a were determined to be 15%, 25% and 45%, respectively. In vitro release of pDNAs from microspheres was evaluated for a 45-day period with no conformational changes and A2 displayed slower release than that of the B2 and B2a. When mice were immunized from anterior tibialis muscle using A2, B2 and B2a formulations containing 100 microg pDNA, antibody responses were detected for 6 months in mice sera. CONCLUSIONS: Although all PLGA microsphere formulations containing pDNA elicited antibody responses by the end of the 6th month, the antibody titers were found to be higher with B2 and B2a formulations in comparison to A2 formulation and the naked pDNA in saline. PMID- 19545225 TI - The intestinal permeation of didanosine from granules containing microspheres using the everted gut sac model. AB - The aim of this research is to evaluate the intestinal permeation of a new formulation (NF) for the anti-retroviral didanosine (ddI) drug, using the everted gut sac model. The NF is composed by granules containing ddI incorporated in chitosan microspheres, plus free chitosan as an excipient. The permeation was evaluated across the three intestinal segments of adult male Wistar rats. The performance of ddI permeation from the NF was compared to the same granules without free chitosan and to buffered ddI tablets as control. The permeations across duodenum were higher than across jejune and ileum. The ddI from NF presented higher permeation and a crescent-shaped profile in duodenum compared to the other formulations. Such effects are provided by the superior mucoadhesiveness to the intestinal membrane and potentialize sustained release properties for NF. These results lead one to consider the novel formulation to be promising for ddI administration by oral route. PMID- 19545226 TI - Cyclosporine-A incorporated cationic solid lipid nanoparticles for ocular delivery. AB - In the present study, Cyclosporine A (CsA) was successfully incorporated into cationic solid lipid nanoparticles (SLN) for ocular application. Physicochemical characterizations of SLNs were analysed in detail during the storage period of 6 months. Due to the better characteristics like smaller particle size (248.00 +/- 0.33 nm) with narrow size distribution (PI = 0.25 +/- 0.00), high zeta potential (50.30 +/- 0.78 mV) and more stable lipid structure, Dynasan 116 structured FD4 (0.1% CsA) formulation was chosen for in vivo studies. Sheep were used in in vivo studies and 200 microL of formulation was applied to sheep' eyes (n = 6) under veterinarian supervision. Samples were collected at pre-determined time intervals and were analysed by enzyme immune assay (EIA). CsA could be detected in both aqueous and vitreous humour samples for 48 h showing the ocular penetration of formulation. Release profiles were not decreased during 48 h indicating controlled and prolonged release of active agent from positively charged SLN formulations due to increased residence time in eyes. Similarities in CsA concentration data showed that inter-individual variance did not influence the ocular penetration of CsA when formulated as SLN. PMID- 19545230 TI - Biological activities of pyrazoline derivatives--a recent development. AB - Pyrazolines are well known and important nitrogen containing 5-membered heterocyclic compounds and various methods have been worked out for their synthesis. Numerous pyrazoline derivatives have been found to possess considerable biological activities, which stimulated the research activity in this field. They have several prominent effects, such as antimicrobial, antimycobacterial, antifungal, antiamoebic, anti-inflammatory, analgesic, antidepressant and anticancer activities. They also possess some potent receptor selective biological activity like Nitric oxide synthase (NOS) inhibitor and Cannabinoid CB1 receptor antagonists activity. 4,5-dihydro-1H- pyrazolines seem to be the most frequently studied pyrazoline type compounds. As a result, a large number of such pyrazolines using different synthetic methods for their preparation have been described in the chemistry literature. The present review provides an insight view to pyrazolines synthesis and its biological activities along with the compilation of recent patents on pyrazolines. PMID- 19545231 TI - Evaluating long-term relationship of protein sequence by use of D-interval conditional probability and its impact on protein structural class prediction. AB - To fix the large and expanding gap between sequence known proteins and structure known proteins, it is important to study on protein structural class prediction (PSCP) for its foundation and usefulness in protein structure analysis. In this paper, the d-interval conditional probability index was proposed to reflect the long-term correlation between amino acids. Based on this index, the impact of residues' long-term relationship on PSCP was analyzed. Two new information theory based algorithms were proposed and were used combining with the long-term information between residues to predict protein structural class (PSC). The dataset 5714 was tested for its low sequence similarity and high reliability. The result showed that the new index was 3-6% higher than traditional index by use of the same algorithms, and the PSCP accuracy was 4-10% improved using the new algorithms. The presented index, algorithms and the long-term relationship of residues on PSCP can be extensively applied in other sequence based protein structure analysis. PMID- 19545232 TI - An overview about biomedical applications of micron and nano size tantalum. AB - Tantalum is obtained from the minerals colombite, tantalite and euxenite. It is greyish silver, heavy and very hard. Tantalum does not react with body fluids and is used to make surgical equipment. Tantalum also does not irritate the body and is used to make surgical sutures as well as implants, such as artificial joints and cranial plates. Bone growth around tantalum covered by calcium phosphate is described by in vitro experimentations. in vivo. Bioactive properties of porous tantalum have been recently developed. Porous tantalum consists of an interconnected porous structure with an average porosity diameter of approximately 400 microm. Hollow spheres with nanometre-to-micrometer dimensions are widely used in different range of applications such as drug delivery carriers and bioreactors. Porous tantalum metal is currently used in orthopaedic for manufacturing of structural component for primary and revision total hip and knee replacements and, more recently, in spine surgery. Good clinical outcomes have been achieved especially in hip revision surgery, using tantalum implants, and promising short term follow up have been reported for knee revision surgery. Some patents for tantalum biomedical applications have been obtained and this article reviews various recent patents on this material. Many patents are developed on tantalum for biomedical application. This paper review some of them with particular interested for biomedical applications in orthopaedic and dentistry. PMID- 19545233 TI - Heparin-coated extracorporeal circulation systems in heart surgery. AB - Despite the progress accomplished in the field of off-pump heart surgery, the vast majority of cardiac operations are still performed with the use of extracorporeal circulation, otherwise known as "heart-lung machine." This valuable tool, however, is connected with various complications, partly deriving from the application of intravenous heparin, necessary for the extracorporeal circuits to function. In order to deal with these complications, which among others include postoperative hemorrhage and systemic inflammatory response, several extracorporeal circulation systems, which contain a heparin-coating on their blood-contacting surfaces, have been developed with patents. The philosophy behind the creation of these systems is that with the controlled absorption and interaction of this heparin with the blood elements, adequate intraoperative anticoagulation with lower doses of systemic heparin and fewer systemic complications can be achieved. The idea of the use of heparin coatings has also been applied in other settings, such as in renal dialysis catheters, ECMO (extracorporeal membrane oxygenation), MECC (minimized extracorporeal circulation) and left ventricle assist devices. PMID- 19545234 TI - Gender differences in ischemic heart disease. AB - Coronary artery disease (CAD) is a leading cause of mortality and morbidity in most developed countries. Gender-related differences have been found in the presentation, prevalence, and clinical outcomes of CAD in many studies. Compared to women, men present with ST-segment elevation myocardial infarction more often and have a higher prevalence of CAD. These findings indicate that gender may have an important influence on CAD. Appropriate diagnosis, prevention, recent patent inventions, and treatment will improve the care of all patients. It is therefore necessary to consider the differences in the features of ischemic heart disease between men and women when examining patients. Novel drugs for tailor-made therapy based on gender differences should be developed for the treatment of CAD in future. PMID- 19545235 TI - Fabrication and physical-chemical characterisation of polyelectrolyte microparticles: platform for controlled release of bioactives. AB - Porous CaCO(3) microparticles were fabricated by colloidal crystallization. Two oppositely charged polyelectrolytes, poly (styrene sulfonate, PSS) and poly (allylamine hydrochloride, PAH) were adsorbed layer-by-layer on the CaCO(3) templates. Polyelectrolyte microcapsules were then obtained by removing the CaCO(3) core. Scanning electron microscopy (SEM), energy-dispersion X-ray analysis (EDX), laser diffraction particle sizing and Raman spectroscopy were employed to characterize the physico-chemical properties of the constructed microcapsules. In vitro drug release studies were conducted using the model water soluble drug Rhodamine B. Factors such as the number of polyelectrolyte layers and pH were investigated. SEM micrographs revealed uniform CaCO(3) microparticles, nearly spherical in shape with pronounced surface roughness, and highly developed interior porous structure. The surface of polyelectrolyte coated particles became rougher than the initial CaCO(3) microparticles. The acquired SEM micrographs of the (PSS/PAH)(n) microcapsules indicated that the number of layers affected the morphology of the microcapsules. The (PSS/PAH)(3) microcapsules revealed a very porous network with many holes resembling the initial morphology of CaCO(3) microparticles. Raman spectra showed peaks at 1125 cm(-1) (S=O bond) and 1600 cm(-1) (aromatic ring stretching) which represented the PSS molecule. The thickness of each layer was about 10 to 20 nm and it can be tailored to such nanometer level by controlling the number of adsorbed layers. The in vitro release of Rhodamine B was dependent on both the number of wall bilayers as well as the pH of the release media. These systems provide an opportunity for the development of controlled release dosage forms with greater effectiveness in the treatment of chronic conditions. PMID- 19545229 TI - Electroporation advances in large animals. AB - In vivo electroporation-mediated gene therapy in large animals is gaining ground as one of the most important means for non-viral gene therapy. This review focuses on the novel aspects of reversible electroporation as applied to large animals, improvement of electroporation delivery technique, and development of electroporation-based vaccines. In regard to large animals, we have summarized the initial use of electroporation-mediated antineoplastic gene therapy in humans, vaccination in monkeys, reversing and preventing cachexia in dogs, and increases growth rate and piglet survival in pigs. Novel techniques incorporating electroporation, including ex vivo manipulations, electron avalanche transfection, and electrosonoporation illustrate evolving modifications. Specific alterations of electroporation parameters and DNA formulations along with ideas of enhancing gene transfection efficiency are provided in addition to a discussion of some of the current limitations of electroporation-mediated gene therapy. PMID- 19545236 TI - The NRAMP6 metal transporter contributes to cadmium toxicity. AB - NRAMP (natural resistance-associated macrophage protein) homologues are evolutionarily conserved bivalent metal transporters. In Arabidopsis, AtNRAMP3 and AtNRAMP4 play a key role in iron nutrition of the germinating plantlet by remobilizing vacuolar iron stores. In the present paper we describe the molecular and physiological characterization of AtNRAMP6. AtNRAMP6 is predominantly expressed in the dry seed embryo and to a lesser extent in aerial parts. Its promoter activity is found diffusely distributed in cotyledons and hypocotyl, as well as in the vascular tissue region of leaf and flower. We show that the AtNRAMP6 transcript coexists with a partially spliced isoform in all shoot cell types tested. When expressed in yeast, AtNRAMP6, but not its misspliced derivative, increased sensitivity to cadmium without affecting cadmium content in the cell. Likewise, Arabidopsis transgenic plants overexpressing AtNRAMP6 were hypersensitive to cadmium, although plant cadmium content remained unchanged. Consistently, a null allele of AtNRAMP6, named nramp6-1, was more tolerant to cadmium toxicity, a phenotype that was reverted by expressing AtNRAMP6 in the mutant background. We used an AtNRAMP6::HA (where HA is haemagglutinin) fusion, shown to be functional in yeast, to demonstrate through immunoblot analysis of membrane fractions and immunofluorescence localization that, in yeast cells, AtNRAMP6 is targeted to a vesicular-shaped endomembrane compartment distinct from the vacuole or mitochondria. We therefore propose that AtNRAMP6 functions as an intracellular metal transporter, whose presence, when modified, is likely to affect distribution/availability of cadmium within the cell. PMID- 19545237 TI - Generation of tumour-necrosis-factor-alpha-specific affibody molecules capable of blocking receptor binding in vitro. AB - Affibody molecules specific for human TNF-alpha (tumour necrosis factor-alpha) were selected by phage-display technology from a library based on the 58-residue Protein A-derived Z domain. TNF-alpha is a proinflammatory cytokine involved in several inflammatory diseases and, to this day, four TNF-alpha-blocking protein pharmaceuticals have been approved for clinical use. The phage selection generated 18 unique cysteine-free affibody sequences of which 12 were chosen, after sequence cluster analysis, for characterization as proteins. Biosensor binding studies of the 12 Escherichia coli-produced and IMAC (immobilized-metal ion affinity chromatography)-purified affibody molecules revealed three variants that demonstrated the strongest binding to human TNF-alpha. These three affibody molecules were subjected to kinetic binding analysis and also tested for their binding to mouse, rat and pig TNF-alpha. For ZTNF-alpha:185, subnanomolar affinity (KD=0.1-0.5 nM) for human TNF-alpha was demonstrated, as well as significant binding to TNF-alpha from the other species. Furthermore, the binding site was found to overlap with the binding site for the TNF-alpha receptor, since this interaction could be efficiently blocked by the ZTNF-alpha:185 affibody. When investigating six dimeric affibody constructs with different linker lengths, and one trimeric construct, it was found that the inhibition of the TNF-alpha binding to its receptor could be further improved by using dimers with extended linkers and/or a trimeric affibody construct. The potential implication of the results for the future design of affibody-based reagents for the diagnosis of inflammation is discussed. PMID- 19545239 TI - Community consequences of herbivore-induced bottom-up trophic cascades: the importance of resource heterogeneity. AB - 1. Herbivory often changes resource quality for herbivorous insects through induced plant responses depending on the intensity of damage. We hypothesized that the willow's response following herbivory affects the entire arthropod community structure on the tree due to changes in plant quality. To examine this hypothesis, we investigated arthropod communities on three willow species, Salix gilgiana, Salix eriocarpa and Salix serissaefolia. 2. In our common garden, cuttings were established in 2003 and experimental treatments were applied in 2005. Damage by a boring caterpillar of a swift moth (Endoclita excrescens) and artificial cutting of 25% stems were applied as partial herbivory within individual trees, and 100% cutting of stems was applied to represent severe herbivory to whole individual trees. These treatments stimulated lateral shoot production depending on damage intensity, resulting in full compensation for biomass loss. 3. Positive relationships were detected between within-tree variation in foliar nitrogen content and overall abundance/species richness of herbivores. Moth boring and 25% cutting increased herbivore abundance and species richness relative to controls. However, we found no significant differences in herbivore abundance and species richness between 100% cut and control trees. Community composition of herbivore species was significantly different between the following three groups: (i) bored and 25% cut; (ii) 100% cut; and (iii) control trees. Changes in community structure of herbivores were likely due to changes in plant quality depending on the intensity of damage. 4. Although total predator abundance and species richness were not significantly different among treatments, community composition of predator species was significantly different among treatments. 5. These results indicate that herbivore-induced willow responses can largely determine the entire arthropod community structure of multitrophic levels due to changes in plant quality. We suggest that heterogeneous resource conditions induced by herbivory within and among plant individuals increase the species diversity of arthropods. PMID- 19545238 TI - Recombinant production of eight human cytosolic aminotransferases and assessment of their potential involvement in glyoxylate metabolism. AB - PH1 (primary hyperoxaluria type 1) is a severe inborn disorder of glyoxylate metabolism caused by a functional deficiency of the peroxisomal enzyme AGXT (alanine-glyoxylate aminotransferase), which converts glyoxylate into glycine using L-alanine as the amino-group donor. Even though pre-genomic studies indicate that other human transaminases can convert glyoxylate into glycine, in PH1 patients these enzymes are apparently unable to compensate for the lack of AGXT, perhaps due to their limited levels of expression, their localization in an inappropriate cell compartment or the scarcity of the required amino-group donor. In the present paper, we describe the cloning of eight human cytosolic aminotransferases, their recombinant expression as His6-tagged proteins and a comparative study on their ability to transaminate glyoxylate, using any standard amino acid as an amino-group donor. To selectively quantify the glycine formed, we have developed and validated an assay based on bacterial GO (glycine oxidase); this assay allows the detection of enzymes that produce glycine by transamination in the presence of mixtures of potential amino-group donors and without separation of the product from the substrates. We show that among the eight enzymes tested, only GPT (alanine transaminase) and PSAT1 (phosphoserine aminotransferase 1) can transaminate glyoxylate with good efficiency, using L glutamate (and, for GPT, also L-alanine) as the best amino-group donor. These findings confirm that glyoxylate transamination can occur in the cytosol, in direct competition with the conversion of glyoxylate into oxalate. The potential implications for the treatment of primary hyperoxaluria are discussed. PMID- 19545240 TI - Professor Robert John Pierce, 15 January 1947--7 February 2009. PMID- 19545241 TI - Increasing complexities of stroke care. PMID- 19545242 TI - The implementation of intravenous tissue plasminogen activator in acute ischaemic stroke--a scientific position statement from the National Stroke Foundation and the Stroke Society of Australasia. AB - Intravenous tissue plasminogen activator (tPA) has been licensed in Australia for thrombolysis in selected patients with acute ischaemic stroke since 2003. The use of tPA is low but is increasing across Australia and national audits indicate efficacy and safety outcomes equivalent to international benchmarks. Implementing tPA therapy in clinical practice is, however, challenging and requires a coordinated multidisciplinary approach to acute stroke care across prehospital, emergency department and inpatient care sectors. Stroke care units are an essential ingredient underpinning safe implementation of stroke thrombolysis. Support systems such as care pathways, therapy delivery protocols, and thrombolysis-experienced multidisciplinary care teams are also important enablers. Where delivery of stroke thrombolysis is being planned, health systems need to be re-configured to provide these important elements. This consensus statement provides a review of the evidence for, and implementation of, tPA in acute ischaemic stroke with specific reference to the Australian health-care system. PMID- 19545243 TI - Acute stroke and transient ischaemic attack management--time to act fast. AB - Stroke is Australia's second single greatest killer with 53 000 new events each year at a rate of 1 every 10 min. Stroke services should be organized to enable people to access proven therapies, such as stroke unit care and thrombolysis, to reduce the impact of stroke. Timely, efficient and coordinated care from ambulance services, emergency services and stroke services will maximize recovery and prevent costly complications and subsequent strokes. Efficient management of patients with transient ischaemic attack can produce significant reductions in subsequent stroke events and risk stratification using the ABCD2 tool can aid management decisions. Evidence for acute stroke care continues to evolve and it is crucial that health professionals are aware of, and implement, best practice clinical guidelines for stroke care. PMID- 19545244 TI - There's more to transient ischaemic attack than ABCD. AB - Several important studies have contributed to recent changes in clinical thinking regarding transient ischaemic attack (TIA). The use of risk stratification scores and urgent clinics has become popular and incorporated into recent guidelines, but there may be some pitfalls with this approach. This commentary stresses the need for careful individual patient assessment focussed on prompt determination of the underlying pathogenic mechanism for the TIA. Suggestions on how to incorporate assessment of TIA patients into the current duties of stroke units are made. PMID- 19545245 TI - Search for pathogenetic variants of the SPRY2 gene in intestinal innervation defects. AB - SPRY2 is an inducible inhibitor of signalling mediated by tyrosine kinases receptors, whose targeting causes intestinal hyperganglionosis in mice. In this light, we have undertaken a mutational analysis of the SPRY2 gene in patients affected with intestinal neuronal dysplasia (IND), without detecting nucleotide changes in any of the 26 DNA samples analysed, with the exception of two already known polymorphic variants. A role of the SPRY2 gene in IND pathogenesis can be thus excluded. PMID- 19545246 TI - Ticlopidine-induced myelosuppression spontaneously remitted after five years: a possible link with amlodipine coadministration. PMID- 19545247 TI - Non-invasive foetal RHD genotyping via real-time PCR of foetal DNA from Chinese RhD-negative maternal plasma. AB - BACKGROUND: A majority of studies predicting the foetal RhD blood group in free foetal DNA from RhD-negative maternal plasma have been conducted in Caucasian populations, whereas limited data have been accumulated for Asian populations. In this study, we assessed the feasibility of prenatal genotyping of RHD in RhD negative Chinese pregnant women. MATERIALS AND METHODS: Cell-free plasma DNA was extracted from 78 RhD-negative Chinese women carrying a singleton foetus (gestation between 14 and 40 weeks). Foetal DNA was confirmed by testing SRY or nine different polymorphic STR loci in the maternal plasma and buffy coat. Foetal RHD exons 5, 7 and 10 and intron 4 were successfully amplified with RQ-PCR. The RHD1227A allele was examined in all RhD-positive individuals. The foetal RHD genotyping results were compared with the infant cord blood serological analysis. RESULTS: Among the 78 specimens, RHD genotyping results of 70 cases were in complete concordance with serological results from foetal umbilical cord blood. Sixty of these cases were identified as RhD-positive, and 10 cases were typed as RhD-negative. In addition, five cases were 'false-positives', while three cases were considered inconclusive. The detection rate was 89.7% (70/78). In four of the five 'false-positive' cases, the RhDel phenotype was assessed by detecting the RHD1227A allele. Thus, this method yielded a 94.9% (74/78) accuracy rate. CONCLUSIONS: The correct foetal RhD phenotype may be accurately predicted from RhD-negative maternal plasma in Chinese subjects. The RHD1227A allele proved to be an important genetic marker in the RhDel Chinese population. PMID- 19545249 TI - A randomized, double blind, placebo-controlled study of MAP0004 in adult patients with migraine. AB - BACKGROUND: Dihydroergotamine mesylate (DHE) is an effective treatment for acute migraine, but its effective use is often limited by the inconvenience and inconsistency of intranasal, intramuscular, or subcutaneous routes of administration. A new formulation of DHE delivered through the lungs by the novel Tempo inhaler is being developed and is designed to offer fast onset, consistent dosing, and sustained response. OBJECTIVE: This proof of principle and dose setting study evaluated the efficacy and tolerability of inhaled DHE delivered by a breath-synchronized, plume-controlled inhaler (Tempo) in adult migraineurs. METHODS: This was a randomized, double blind, placebo-controlled, 2-period study conducted at 9 headache centers in the United States. Adult men and women with a documented history of acute migraine for at least 12 months, with an average of 2 to 8 attacks per month in the preceding 6 months were treated with MAP0004 0.5 or 1.0 mg systemic equivalent dose (1.0 or 2.0 mg nominal dose) or matching placebo during Treatment Period 1 (TP1). Patients who responded to treatment during TP1 were re-randomized in Treatment Period 2 (TP2) to receive MAP0004 0.25 mg systemic equivalent dose or placebo. RESULTS: Of 86 patients randomized to treatment, 69 were included in the As-Treated population in TP1. Pain relief at 2 hours was greater for MAP0004 0.5 mg (72%, P = .019) and 1.0 mg (65%, P = .071) than for placebo (33%). Pain relief at 10 (32%), 15 (46%), and 30 (55%) minutes was significantly (P < .05) greater with MAP0004 0.5 mg than with placebo (0%, 7% and 14%, respectively). Pain-free at 2 hours was significantly greater with MAP0004 0.5 mg (44%, P = .015) and 1.0 mg (35%, P = .050) than with placebo (7%). Total migraine relief at 2 hours was significantly (P = .019) greater with MAP0004 0.5 mg (72%) than with placebo (33%). Sustained pain relief and pain-free rates exhibited a therapeutic gain of 30% (P = .066) and 31% (P = .037) at 24 hours and 28% (P = .096) and 30% (P = .057) at 48 hours with MAP0004 0.5 mg vs placebo. MAP0004 was well tolerated with no serious or severe adverse events. Dysgeusia was reported as treatment-related in 2 patients on placebo, 0 patients on MAP0004 0.5 mg, and 6 patients on MAP0004 1.0 mg. No clinically relevant changes were noted in spirometry, vital signs, electrocardiogram, or clinical laboratory values. No significant differences between treatments were observed in TP2. CONCLUSIONS: In this study MAP0004 0.5 mg and 1.0 mg were well tolerated and effective at delivering clinically significant, rapid, and sustained pain relief in adult migraine patients. No additional benefit was observed with the higher dose, thus the MAP0004 0.5 mg systemic equivalent dose has been selected as the dose for further clinical study. PMID- 19545250 TI - The effect of pregnancy and parity on headache prevalence: the Head-HUNT study. AB - BACKGROUND: In previous studies, pregnancy has been associated with less headache, but the influence of parity on this association is largely unknown. OBJECTIVES: To examine the prevalence of headache and migraine among pregnant women, and explore the relation of headache to parity in a large, population based study. METHODS: In the Nord-Trondelag Health Study in Norway 1995-1997 (HUNT 2), a total of 27,700 (60%) out of 46,506 invited women responded to headache questions (Head-HUNT). In total, 9281 women were 40 years or younger and responded to questions on pregnancy and birth, and 550 of these reported to be pregnant when filling in the questionnaire. A total of 20,287 women who were 70 years or younger and reported not to be pregnant responded to questions on headache and reported number of child births. RESULTS: Adjusting for age and educational level, the headache prevalence was lower among pregnant than among nonpregnant women. The association between headache and pregnancy was significant for nulliparous (one who has never given birth) (OR = 0.5, 95% CI = 0.4-0.7), but not for primiparous (1 child only) and multiparous (several children) women (OR = 0.8, 95% CI = 0.7-1.0). This was evident for both migraine and nonmigrainous headache. Among nonpregnant women, there was an increased headache prevalence among primi- and multiparous women compared with nulliparous (OR = 1.3, 95% CI = 1.2-1.4). CONCLUSION: Headache, both migraine and nonmigrainous, was less prevalent in nulliparous pregnant women compared with all nonpregnant women, and to nulliparous nonpregnant women. Headache was less prevalent in the third trimester of pregnancy, but not in the first and second trimesters, compared with nonpregnant women. Also in nonpregnant women, headache was less prevalent in nulliparous than in primi- and multiparous women. PMID- 19545252 TI - Headache at onset of acute cerebral ischemia. PMID- 19545254 TI - Postural headache. PMID- 19545255 TI - Migraine and the environment. AB - Migraineurs often describe environmental triggers of their headaches, such as barometric pressure change, bright sunlight, flickering lights, air quality, and odors. Environmental aspects of indoor space and workplaces are also implicated in migraine experience. Comprehensive migraine treatment programs emphasize awareness and avoidance of trigger factors as part of the therapeutic regimen. As migraine has a substantial economic impact, remediation of correctable environmental triggers may benefit employee attendance and productivity among migraineurs. Few controlled studies in the literature, however, confirm environmental influences on migraine and headaches. Although some are controversial, migraineurs worldwide consistently report similar environmental triggers. This article addresses commonly mentioned environmental triggers with a discussion of their pathophysiology and proposed preventive measures. OBJECTIVE: To examine the epidemiological evidence for commonly-mentioned environmental migraine triggers, discuss their possible role in the pathophysiology of migraine and propose preventive measures to avoid or minimize exposure. BACKGROUND: Migraineurs often describe environmental triggers of their headaches, such as barometric pressure change, bright sunlight, flickering lights, air quality and odors. Environmental aspects of indoor space and workplaces are also implicated in the migraine experience. As migraine has a substantial economic impact, remediation of correctable environmental triggers may improve attendance and productivity among migraineurs in the workplace. METHODS: We reviewed the literature addressing indoor and outdoor environmental factors which are commonly implicated as migraine triggers. RESULTS: Although some factors are controversial, migraineurs worldwide consistently report similar environmental triggers. However, few studies confirm environmental influences on migraine and headaches. Research to date indicates that migraineurs have lower thresholds for light-induced discomfort, sine grating distortion and illusions, noise tolerance and olfactory sensitivity compared to the general population. CONCLUSION: There are conflicting studies supporting the validity of patient-reported environmental migraine triggers. Prospective studies are needed to determine the extent that external stimuli influence the migraine process. Decreased thresholds for light, noise, olfactory and visual stimuli in migraineurs may be minimized by modifying the work, home and classroom settings. PMID- 19545256 TI - A critical view on the role of migraine triggers in the genesis of migraine pain. AB - A number of distinct endogenous and exogenous factors have been implicated in migraine precipitation but the exact nature of the triggering process itself and its relationship to the genesis of the headache remains largely speculative. In this article, we examine the potential sites and downstream cascades through which migraine triggers might exert their action to promote the activation of the migraine pain pathway. We further look at the laterality of the headache as a potential indicator for the site of migraine pain initiation and examine the question of triggering factor specificity in relation to the current understanding of migraine pathophysiology. PMID- 19545257 TI - Migraine triggers. PMID- 19545258 TI - Margaritas... not science. PMID- 19545260 TI - Headache prevention with complementary and alternative medicine. PMID- 19545262 TI - Environment specific pleiotropy facilitates divergence at the Ectodysplasin locus in threespine stickleback. AB - Adaptive radiation occurs when divergent natural selection in different environments leads to phenotypic differentiation. The pleiotropic effects of underlying genes can either promote or constrain this diversification. Identifying the pleiotropic effects of genes responsible for divergent traits, and testing how the environment influences these effects, can therefore help to provide an understanding of how ecology drives evolutionary change between populations. Positive selection on low-armor alleles at the Ectodysplasin (Eda) locus in threespine stickleback has led to the repeated evolution of reduced armor in populations following freshwater colonization by fully armored marine sticklebacks. Here, we demonstrate that Eda has environmentally determined pleiotropic effects on armor and growth. When raised in freshwater, reduced armor sticklebacks carrying "low" alleles at Eda had increased growth rate relative to fully armored sticklebacks carrying "complete" alleles. In saltwater treatments this growth advantage was present during juvenile growth but lost during adult growth, suggesting that in this environment stickleback are able to develop full armor plates without sacrificing overall growth rate. The environment specific pleiotropic effects of Eda demonstrate that ecological factors can mediate the influence of genetic architecture in driving phenotypic evolution. Furthermore, because size is important for mate choice in stickleback, the growth rate differences influenced by Eda may have effects on reproductive isolation between marine and freshwater populations. PMID- 19545261 TI - Inhaled house dust mite induces pulmonary T helper 2 cytokine production. AB - BACKGROUND: Inhaled house dust mite (HDM) results in T-helper (TH) 2 type pathology in unsensitized mice, in conjunction with airway hyperreactivity and airway remodelling. However, the pulmonary cytokine and chemokine profile has not been reported. METHODS: We have performed a time course analysis of the characteristic molecular mediators and cellular influx in the bronchoalveolar lavage (BAL) and lung in order to define the pulmonary inflammatory response to inhaled HDM extract. Mice were exposed five times a week to soluble HDM extract for 3 weeks. Lung function was measured in groups of mice at intervals following the final HDM challenge. Recruitment of inflammatory cells and inflammatory mediator production was then assessed in BAL and lungs of individual mice. RESULTS: We found that Th2 cytokines were significantly increased in BAL and lung after HDM challenge from as early as 2 h post-final challenge. The levels of cytokines and chemokines correlated with the influx of eosinophils and Th2 cells to the different compartments of the lung. However, the production of key cytokines such as IL-4, IL-5 and IL-13 preceded the increase in airways resistance. CONCLUSION: Inhaled HDM challenge induces a classical Th2 inflammatory mediator profile in the BAL and lung. These data are important for studies determining the efficacy of novel treatment strategies for allergic airways disease. PMID- 19545263 TI - Metamodels and phylogenetic replication: a systematic approach to the evolution of developmental pathways. AB - Molecular genetic analysis of phenotypic variation has revealed many examples of evolutionary change in the developmental pathways that control plant and animal morphology. A major challenge is to integrate the information from diverse organisms and traits to understand the general patterns of developmental evolution. This integration can be facilitated by evolutionary metamodels-traits that have undergone multiple independent changes in different species and whose development is controlled by well-studied regulatory pathways. The metamodel approach provides the comparative equivalent of experimental replication, allowing us to test whether the evolution of each developmental pathway follows a consistent pattern, and whether different pathways are predisposed to different modes of evolution by their intrinsic organization. A review of several metamodels suggests that the structure of developmental pathways may bias the genetic basis of phenotypic evolution, and highlights phylogenetic replication as a value-added approach that produces deeper insights into the mechanisms of evolution than single-species analyses. PMID- 19545264 TI - Correlated evolution of mating strategy and inbreeding depression within and among populations of the hermaphroditic snail Physa acuta. AB - Inbreeding depression is one of the main forces opposing the evolution of self fertilization. Of central importance is the hypothesis that inbreeding depression and selfing coevolve antagonistically, generating either low selfing rate and high inbreeding depression or vice versa. However, there is limited evidence for this coevolution within species. We investigated this topic in the hermaphroditic snail Physa acuta. In this species, isolated individuals delay the onset of egg laying compared to individuals having access to mates. Longer delays ("waiting times") indicate more intense selfing avoidance. We measured inbreeding depression and waiting time in a large quantitative-genetic experiment (281 outbred families derived from 26 natural populations). We observed large genetic variance for both traits and a strong positive genetic covariance between them, most of which resided within rather than among populations. It means that, within populations, individuals with higher mutation load avoided selfing more strongly on average. This genetic covariance may result from pleiotropy and/or linkage disequilibrium. Whatever its genetic architecture, the fact it emerges specifically when individuals are deprived of mates suggests it is not fortuitous and rather reflects the action of natural selection. We conclude that a diversity of mating strategies can arise within populations subjected to variation in inbreeding depression. PMID- 19545265 TI - Asymmetry of genetic variation in fitness-related traits: apparent stabilizing selection on g(max). AB - The maintenance of genetic variation in traits closely associated with fitness remains a key unresolved issue in evolutionary genetics. One important qualification on the observation of genetic variation in fitness-related traits is that such traits respond asymmetrically to selection, evolving to a greater extent in the direction of lower fitness. Here we test the hypothesis that standing genetic variation in fitness-related traits is principally maintained for unfit phenotypes. Male Drosophila bunnanda vary in mating success (the primary determinant of male fitness) due to female mate choice. We used competitive mating success to partitioning males into two groups: successful (high fitness) and unsuccessful (low fitness). Relative to successful males, unsuccessful males harbored considerably greater levels of additive genetic variation for sexual signaling traits. This genetic asymmetry was detected for a multivariate trait that we demonstrated was not directly under stabilizing sexual selection, leading us to conclude the trait was under apparent stabilizing selection. Consequently, our results suggest genetic variance might be biased toward low fitness even for traits that are not themselves the direct targets of selection. Simple metrics of genetic variance are unlikely to be adequate descriptors of the complex nature of the genetic basis of traits under selection. PMID- 19545266 TI - Fitness trade-offs modify community composition under contrasting disturbance regimes in Pseudomonas fluorescens microcosms. AB - Disturbance is thought to be a major factor influencing patterns of biodiversity. In addition, disturbance can modify community composition if there are species specific trade-offs between fitness and disturbance tolerance. Here, we examine the role of disturbance on the evolution of coexisting biofilm-forming morphotypes of Pseudomonas fluorescens maintained in spatially structured laboratory microcosms. We identified four heritably stable ecotypes that varied significantly in their competitiveness under different disturbance treatments. Furthermore, we identified significant trade-offs in competitiveness across disturbance treatments for three of four of these ecotypes. These trade-offs modified dominance relationships between strains and thus altered community composition, with a peak of ecotype diversity occurring at intermediate disturbance frequencies. PMID- 19545267 TI - Asymmetric, bimodal trade-offs during adaptation of Methylobacterium to distinct growth substrates. AB - Trade-offs between selected and nonselected environments are often assumed to exist during adaptation. This phenomenon is prevalent in microbial metabolism, where many organisms have come to specialize on a narrow breadth of substrates. One well-studied example is methylotrophic bacteria that can use single-carbon (C(1)) compounds as their sole source of carbon and energy, but generally use few, if any, multi-C compounds. Here, we use adaptation of experimental populations of the model methylotroph, Methylobacterium extorquens AM1, to C(1) (methanol) or multi-C (succinate) compounds to investigate specialization and trade-offs between these two metabolic lifestyles. We found a general trend toward trade-offs during adaptation to succinate, but this was neither universal nor showed a quantitative relationship with the extent of adaptation. After 1500 generations, succinate-evolved strains had a remarkably bimodal distribution of fitness values on methanol: either an improvement comparable to the strains adapted on methanol or the complete loss of the ability to grow on C(1) compounds. In contrast, adaptation to methanol resulted in no such trade-offs. Based on the substantial, asymmetric loss of C(1) growth during growth on succinate, we suggest that the long-term maintenance of C(1) metabolism across the genus Methylobacterium requires relatively frequent use of C(1) compounds to prevent rapid loss. PMID- 19545268 TI - Case studies and mathematical models of ecological speciation. 4. Hybrid speciation in butterflies in a jungle. AB - We build a spatial individual-based multilocus model of homoploid hybrid speciation tailored for a tentative case of hybrid origin of Heliconius heurippa from H. melpomene and H. cydno in South America. Our model attempts to account for empirical patterns and data on genetic incompatibility, mating preferences and selection by predation (both based on coloration patterns), habitat preference, and local adaptation for all three Heliconius species. Using this model, we study the likelihood of recombinational speciation and identify the effects of various ecological and genetic parameters on the dynamics, patterns, and consequences of hybrid ecological speciation. Overall, our model supports the possibility of hybrid origin of H. heurippa under certain conditions. The most plausible scenario would include hybridization between H. melpomene and H. cydno in an area geographically isolated from the rest of both parental species with subsequent long-lasting geographic isolation of the new hybrid species, followed by changes in the species ranges, the secondary contact, and disappearance of H. melpomene-type ecomorph in the hybrid species. However, much more work (both empirical and theoretical) is necessary to be able to make more definite conclusions on the importance of homoploid hybrid speciation in animals. PMID- 19545269 TI - Testing phylogeographic hypotheses in a Euro-Siberian cold-adapted leaf beetle with coalescent simulations. AB - Few studies to date have investigated the impact of Pleistocene climatic oscillations on the genetic diversity of cold-adapted species. We focus on the geographic distribution of genetic diversity in a Euro-Siberian boreo-montane leaf beetle, Gonioctena pallida. We present the molecular variation from three independent gene fragments over the entire geographic range of this insect. The observed sequence variation identifies a genetic diversity hot spot in the Carpathian Mountains, in central Europe, which reveals the presence of (1) an ancestral refuge population or (2) a secondary contact zone in this area. Modeling of population evolution in a coalescent framework allowed us to favor the ancestral refuge hypothesis. These analyses suggest that the Carpathian Mountains served as a refuge for G. pallida, whereas the rest of the species distribution, that spans a large portion of Europe and Asia, experienced a dramatic reduction in genetic variation probably associated to bottlenecks and/or founder events. We estimated the time of isolation of the ancestral refuge population, using an approximate Bayesian method, to be larger than 90,000 years. If true, the current pattern of genetic variation in this cold-adapted organism was shaped by a climatic event predating by far the end of the last ice age. PMID- 19545271 TI - Clinical guidelines: a sword or a shield in clinical negligence litigation? PMID- 19545272 TI - Baseline renal function, ischaemia time and blood loss predict the rate of renal failure after partial nephrectomy. AB - OBJECTIVE: To identify independent predictors of renal failure after partial nephrectomy (PN) in patients with renal cell carcinoma (RCC). PATIENTS AND METHODS: Data were available for 166 patients with pathological T1-3 N0M0 RCC treated with PN. Renal failure after PN was defined as a decrease in glomerular filtration rate (GFR) of >25% (RIFLE criteria). The GFR before and after PN was estimated using the Modification of Diet in Renal Disease study group equation. Univariable and multivariable logistic regression models were used to assess a decrease of >25% in GFR from the preoperative level. Candidate predictor variables were age, gender, PN indication (absolute vs relative), preoperative GFR, tumour size, perioperative blood loss, surgery duration and clamping time. RESULTS: After PN, 22 (13.3%) patients had a decrease in GFR of >25%. The perioperative blood loss (P = 0.02), clamping time (P = 0.04) and preoperative GFR (P = 0.002) were independent predictors of a decrease in GFR of >25%. CONCLUSIONS: We identified two important potentially modifiable variables that should be considered in the planning of PN, i.e. the clamping time and blood loss. It is possible that selective referral to experienced surgeons who can perform PN within short surgical and clamping times, and with minimal blood loss, could minimize the rate of renal failure, especially in patients with an underlying renal function impairment. PMID- 19545274 TI - Re: A second cycle of tamsulosin in patients with distal ureteric stones: a prospective randomized trial. PMID- 19545276 TI - The impact of vector-mediated neutrophil recruitment on cutaneous leishmaniasis. AB - The dynamic process of pathogen transmission by the bite of an insect vector combines several biological processes that have undergone extensive co-evolution. Whereas the host response to an insect bite is only occasionally confronted with the parasitic pathogens that competent vectors might transmit, the transmitted parasites will always be confronted with the acute, wound-healing response that is initiated by the bite itself. Invariably, this response involves neutrophils. In the case of Leishmania, infection is initiated in the skin following the bite of an infected sand fly, suggesting that Leishmania must possess some means to survive their early encounter with recruited neutrophils at the bite site. Here, we review the literature regarding the impact of neutrophils on the outcome of infection with Leishmania, with special attention to the role of the sand fly bite. PMID- 19545277 TI - Leucine-rich repeat kinase 2 phosphorylates brain tubulin-beta isoforms and modulates microtubule stability--a point of convergence in parkinsonian neurodegeneration? AB - Autosomal dominant mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common genetic cause of late-onset Parkinson's disease. The most prevalent LRRK2(G2019S) mutation has repeatedly been shown to enhance kinase activity and neurotoxicity, however, the molecular mechanisms leading to neurodegeneration remain poorly defined. Here we show that recombinant human LRRK2 preferentially phosphorylates tubulin-beta purified from bovine brain and that phosphorylation is three-fold enhanced by the LRRK2(G2019S) mutation. By tandem mass spectrometry, Thr107 was identified as phosphorylation site which is highly conserved between tubulin-beta family members and also between tubulin-beta genes of different species. LRRK2 was co-immunoprecipitated with tubulin-beta both from wild-type mouse brain and from LRRK2 over-expressing, non-neuronal human embryonic kidney 293 cells. However, an effect of LRRK2 on tubulin phosphorylation and assembly was only detectable in mouse brain samples. In vitro co-incubation of bovine brain tubulins with LRRK2 increased microtubule stability in the presence of microtubule-associated proteins which may explain the reduction in neurite length in LRRK2-deficient neurons in culture. These findings suggest that LRRK2(G2019S)-induced neurodegeneration in Parkinsonian brains may be partly mediated by increased phosphorylation of tubulin-beta and constraining of microtubule dynamics. PMID- 19545278 TI - Morphine withdrawal regulates phosphorylation of cAMP response element binding protein (CREB) through PKC in the nucleus tractus solitarius-A2 catecholaminergic neurons. AB - The transcription factor cAMP response element binding protein (CREB) has been implicated in the actions of drugs of abuse in several brain areas. However, little is known about CREB regulation in the nucleus tractus solitarius (NTS) A(2) catecholaminergic cell group, one of the key regions of the brain stress system. Morphine withdrawal modulates gene expression in the NTS through various second-messenger signal transduction systems including activation of extracellular signal-regulated kinases 1/2 (ERK(1/2)) and protein kinase C (PKC). In the current study we used immunoblotting and immunohistochemistry to investigate changes in CREB phosphorylation in the NTS and kinases that may mediate the morphine withdrawal-triggered activation of CREB and hypothalamo pituitary-adrenocortical (HPA) axis (another stress system circuit) response after naloxone-induced morphine withdrawal. We found an increased phosphorylation of CREB (pCREB) selectively within tyrosine hydroxylase (TH) immunoreactive neurons in the NTS from morphine-withdrawn rats, which parallel elevated corticosterone levels. We also measured expression levels of TH and phosphorylated ERK(1/2) (pERK(1/2)), and found that both are up-regulated following morphine withdrawal. SL327, an inhibitor of ERK activation, at doses which reduced the hyperactive pERK(1/2) levels, did not attenuated the rise in pCREB and TH immunoreactivity or plasma corticosterone secretion during morphine withdrawal, indicating that ERK kinase/ERK pathway was not directly needed for either activation of CREB and TH expression in the NTS or HPA axis hyperactivity. In contrast, PKC inhibitor calphostin C reduced the withdrawal-triggered rise in pCREB, pERK(1/2), TH expression and corticosterone secretion. The results indicate that PKC mediates both CREB activation and HPA response by morphine withdrawal and might suggest that CREB activation in the NTS is related to TH expression associated with morphine withdrawal. PMID- 19545279 TI - Tryptophan is a marker of human postmortem brain tissue quality. AB - Postmortem human brain tissue is widely used in neuroscience research, but use of tissue originating from different brain bank centers is considered inaccurate because of possible heterogeneity in sample quality. There is thus a need for well-characterized markers to assess the quality of postmortem brain tissue. Toward this aim, we determined tryptophan (TRP) concentrations, phosphofructokinase-1 and glutamate decarboxylase activities in 119 brain tissue samples. These neurochemical parameters were tested in samples from autopsied individuals, including control and pathological cases provided by 10 different brain bank centers. Parameters were assessed for correlation with agonal state, postmortem interval, age and gender, brain region, preservation and freezing methods, storage conditions and storage time, RNA integrity, and tissue pH value. TRP concentrations were elevated significantly (p = 0.045) with increased postmortem interval; which might indicate increased protein degradation. Therefore, TRP concentration might be one useful and convenient marker for estimating the quality of human postmortem brain tissue. PMID- 19545280 TI - Stress hormones collaborate to induce lymphocyte apoptosis after high level spinal cord injury. AB - Post-traumatic immune suppression renders individuals with spinal cord injury (SCI) susceptible to infection. Normally, proper immune function is regulated by collaboration between the sympathetic nervous system (SNS) and hypothalamic pituitary-adrenal (HPA) axis and involves the controlled release of glucocorticoids (GCs) and norepinephrine (NE). Recently, we showed that after high thoracic (T3) SCI, aberrant levels of GCs and NE accumulate in the blood and spleen, respectively. These changes are associated with splenic atrophy, splenic leucopenia, increased intrasplenic caspase 3 levels, and suppressed B lymphocyte function. As GCs boost SNS function, in part by increasing the expression and affinity of beta2 adrenergic receptors (beta2ARs) while simultaneously preventing beta2AR down-regulation, we predicted that surges in stress hormones (i.e., GCs and NE) in the blood and spleen of mice with high-level SCI would act concurrently to adversely affect lymphocyte function and survival. Here, we show that post-SCI concentrations of GCs enhance the sensitivity of lymphocytes to beta2AR stimulation causing an increase in intracellular Bcl-2 interacting mediator of cell death (Bim) and subsequent apoptosis. In vivo, the combined antagonism of GC receptors and beta2ARs significantly diminished lymphocyte Bim levels and SCI-induced splenic lymphopenia. Together, these data suggest that pharmacological antagonists of the HPA/SNS axes should be considered as adjunct therapies for ameliorating post-traumatic immune suppression in quadriplegics and high paraplegics. PMID- 19545283 TI - Successful pregnancy after arsenic trioxide therapy for relapsed acute promyelocytic leukaemia. PMID- 19545281 TI - Oxidized phosphatidylcholine formation and action in oligodendrocytes. AB - Reactive oxygen species play a major role in neurodegeneration. Increasing concentrations of peroxide induce neural cell death through activation of pro apoptotic pathways. We now report that hydrogen peroxide generated sn-2 oxidized phosphatidylcholine (OxPC) in neonatal rat oligodendrocytes and that synthetic OxPC [1-palmitoyl-2-(5'-oxo)valeryl-sn-glycero-3 phosphorylcholine, POVPC] also induced apoptosis in neonatal rat oligodendrocytes. POVPC activated caspases 3 and 8, and neutral sphingomyelinase (NSMase) but not acid sphingomyelinase. Downstream pro-apoptotic pathways activated by POVPC treatment included the Jun N terminal kinase proapoptotic cascade and the degradation of phospho-Akt. Activation of NSMase occurred within 1 h, was blocked by inhibitors of caspase 8, increased mainly C18 and C24:1 ceramides, and appeared to be concentrated in detergent-resistant microdomains (Rafts). We concluded that OxPC initially activated NSMase and converted sphingomyelin into ceramide to mediate a series of downstream pro-apoptotic events in oligodendrocytes. PMID- 19545284 TI - Distribution of NPM1-ALK and X-ALK fusion transcripts in paediatric anaplastic large cell lymphoma: a molecular-histological correlation. AB - Anaplastic large cell lymphomas (ALCL) in children express anaplastic lymphoma kinase (ALK) fusion genes, most commonly NPM1-ALK. The distribution of X-ALK among 66 childhood ALCLs was analysed. One ALCL was ALK-negative. Reverse transcription polymerase chain reaction detected NPM1-ALK in 58 tumours, all showing nuclear and cytoplasmic ALK staining. The remaining seven ALCL stained for ALK in the cytoplasm only: two expressed TPM3-ALK, one ATIC-ALK, one MYH9 ALK; three no TPM3-, TFG-, ATIC-, CLTC- or MYH9-ALK. Almost 90% of paediatric ALK positive ALCLs express NPM1-ALK. There was complete concordance between ALK staining pattern and the presence of a typical/variant ALK fusion partner. PMID- 19545285 TI - Interferon-alpha may restore sensitivity to tyrosine-kinase inhibitors in Philadelphia chromosome positive acute lymphoblastic leukaemia with F317L mutation. PMID- 19545287 TI - Bone marrow metastasis in gastro-intestinal stromal tumour. PMID- 19545286 TI - PEG-IFN-alpha-2a therapy in patients with myelofibrosis: a study of the French Groupe d'Etudes des Myelofibroses (GEM) and France Intergroupe des syndromes Myeloproliferatifs (FIM). PMID- 19545288 TI - Alemtuzumab in the reversal of encephalopathy associated with hypereosinophilic syndrome. PMID- 19545289 TI - Rumpel-Leede sign in thrombocytopenia due to Epstein-Barr virus-induced mononucleosis. PMID- 19545290 TI - Expanded safety experience with lenalidomide plus dexamethasone in relapsed or refractory multiple myeloma. AB - Lenalidomide gained Food and Drug Administration (FDA) approval for treatment of patients with relapsed or refractory multiple myeloma (MM) in combination with dexamethasone in June 2006. In April 2005, the FDA and patient advocacy groups requested an expanded access programme to both provide lenalidomide to patients likely to benefit and obtain additional safety information. Relapsed/refractory MM patients received lenalidomide 25 mg/d (days 1-21) and dexamethasone 40 mg/d (days 1-4, 9-12, and 17-20 of cycles 1-4; days 1-4 only from cycle 5 onwards), in 4-week cycles until disease progression, study drug discontinuation, or lenalidomide approval. Of the 1438 patients enrolled, approximately 60% were male, median age was 64 years, and 61.7% had Durie-Salmon stage III disease. Median time on study was 15.4 weeks (range: 0.1-49.1) and median dose was 25 mg. The most common adverse events (AEs) were haematological (49%), gastrointestinal (59%), and fatigue (55%). The most common grade > or =3 AEs were haematological (45%), fatigue (10%), and pneumonia (7%). The most common serious AEs were pneumonia (8%), pyrexia (4%), and deep-vein thrombosis (3%). Primary cause of death was disease progression (10%). Safety data confirmed known AEs of lenalidomide plus dexamethasone therapy in patients with relapsed/refractory MM. PMID- 19545291 TI - Intra-vitreal methotrexate leads to resolution of intraocular chronic lymphocytic leukaemia. PMID- 19545292 TI - Dismal long-term prognosis for children with refractory acute myeloid leukaemia treated with gemtuzumab ozogamicin and stem cell transplantation: where now? PMID- 19545294 TI - A call for full ingredient labelling and uniform nomenclature on medicaments. PMID- 19545295 TI - Nonbacterial purpura fulminans and severe autoimmune acquired protein S deficiency associated with human herpesvirus-6 active replication. AB - Nonbacterial purpura fulminans (PF) is rare, usually follows viral infection in young children, and is characterized by specific coagulation disorders, requiring specific therapy. Following a transient rash, a 2-year-old previously healthy girl developed PF without haemodynamic impairment. Laboratory data revealed disseminated intravascular coagulation and a severe transient protein S deficiency. Antiprotein S autoantibodies and active human herpesvirus-6 (HHV6) replication were demonstrated. Purpuric skin lesions spread very rapidly despite broad-spectrum antibiotics and right leg amputation. Plasmapheresis and intravenous immunoglobulins gave complete clinical recovery and normalization of protein S level within 10 days, with progressive clearance of antiprotein S autoantibodies. Transient severe protein S deficiencies have previously been reported in patients with nonbacterial PF, usually after varicella infection. This is the first documented case of PF after HHV6 infection. PMID- 19545296 TI - Comments on 'Validity and responsiveness of the Osnabruck Hand Eczema Severity Index (OHSI): a methodological study'. PMID- 19545298 TI - Prevalence and risk factors of inflammatory acne vulgaris in rural and urban Ghanaian schoolchildren. PMID- 19545297 TI - Effect of dosing frequency on the safety and efficacy of imiquimod 5% cream for treatment of actinic keratosis on the forearms and hands: a phase II, randomized placebo-controlled trial. AB - BACKGROUND: Clinical studies in cutaneous conditions other than actinic keratosis (AK) have revealed that the safety and efficacy profile of imiquimod is influenced by dosing frequency. OBJECTIVES: To evaluate dosing frequency response of imiquimod 5% for treatment of AK. METHODS: This was a phase II, multicentre, randomized, double-blind, placebo-controlled study. Adults with > or = 10 but < or = 50 clinical AKs, one of which was histologically confirmed, were randomized (4:1) to 2-6 packets of imiquimod or placebo cream applied to the dorsum of the forearms and hands once daily 2, 3, 5 or 7 times per week for 8 weeks. The primary endpoint was complete clearance of AKs in the treatment area at 8 weeks post-treatment. RESULTS: One hundred and forty-nine (94 men and 54 women) white subjects, with a mean +/- SD age of 71 +/- 10.2 years, were enrolled. Twenty eight subjects (18.8%) discontinued from study: 0%, 3.1%, 6.9%, 30.0% and 33.3% withdrew for local skin reactions or adverse events in the combined placebo, and in the imiquimod 2, 3, 5 or 7 times per week groups, respectively. Seven serious adverse events occurred; none was related to the study drug. Median baseline lesions ranged from 38 to 40 for the treatment groups. Complete clearance was achieved in 0%, 3.2%, 6.9%, 3.3% and 6.7% of subjects, and partial clearance (> or = 75% lesion reduction) in 0%, 22.6%, 24.1%, 20.0% and 36.7% of subjects for the placebo and imiquimod 2, 3, 5 or 7 times per week regimens, respectively. CONCLUSIONS: Imiquimod 5% applied more frequently than 3 times per week to AKs was not well tolerated. Complete clearance rates were low; however, partial clearance rates increased with increased dosing frequency (P = 0.002). PMID- 19545299 TI - Partial nasal reconstruction with titanium mesh: report of five cases. AB - The nasal pyramid is frequently affected by nonmelanoma skin cancer. Sometimes the aggressiveness of tumours entails the extirpation of the mucosa, the cartilage, and the nasal skin. Reconstruction of the cartilaginous portion can be a surgical challenge. We demonstrate that titanium mesh can be an effective substitute for the cartilaginous portion of the nose in nasal reconstruction. We present five patients with nasal basal cell carcinoma who were treated by Mohs micrographic surgery. The partial loss of the cartilaginous structure was replaced by a 0.1 mm fenestrated titanium mesh. We have not observed any rejection or other complication in any of our patients. Good functional and aesthetic results have been obtained. Because of its biocompatibility, titanium mesh is a useful substitute for nasal cartilage. It avoids harvesting natural cartilage, reduces the risk of graft necrosis, and prevents morbidity in the donor area. PMID- 19545318 TI - Colonisation of soft lining materials by micro-organisms. AB - OBJECTIVE: This study evaluated the in vitro adherence of pathogenic micro organisms, Candida albicans, Staphylococcus aureus and Pseudomonas aeruginosa, to soft lining materials and their inhibitory effect on these micro-organisms. MATERIALS AND METHODS: To measure adherence, specimens of Molloplast B and Ufi Gel P were inoculated [10(7 )colony-forming units per millimetre (cfu/ml)] with TSB media containing the micro-organisms. To determine the number of micro organisms in the 10(-2)-10(-5) dilutions, 25 microl of the suspension were transferred to plates of selective media. Colony counts of each specimen were quantified (cfu/ml). The surface roughness was measured with a perfilometer to assess the relationship between the adherence of micro-organisms and surface roughness of each material. For the inhibition test, specimens of materials were placed in agar plates inoculated individually with the micro-organisms. After 48 h, the inhibition zones around the specimens were measured. RESULTS: None of the materials exhibited inhibition zones. The number of cfu/ml of S. aureus and P. aeruginosa were significantly greater than C. albicans for both materials. The Ufi Gel P exhibited greater adherence of C. albicans than Molloplast B. No correlation was observed between the adherence of micro-organisms and surface roughness. CONCLUSION: The surface roughness of the materials is not the only factor governing micro-organism adherence. PMID- 19545319 TI - Denture stomatitis and its risk indicators in south Brazilian older adults. AB - OBJECTIVE: The purpose of this study was to evaluate if socio-demographic, behaviour and health variables are risk indicators for denture-related stomatitis in community-dwelling older adults from Carlos Barbosa, Brazil. MATERIALS AND METHODS: In this cross-sectional study, 872 subjects aged 60 years or more were evaluated. Data collection included dental examinations and questionnaires to assess socio-demographic, behaviour, general and oral health data. All questionnaires and examinations were carried out during 2004. Associations between the independent variables and the outcomes 'prevalence of denture stomatitis' or 'prevalence of granular stomatitis' were adjusted by means of multivariate logistic regressions. RESULTS: Participants with denture stomatitis were significantly more likely to be females, to display increased amounts of plaque, and significantly less likely to visit the dentist for treatment. Participants with granular stomatitis were significantly more likely to be females and participants with granular stomatitis were significantly more likely to be females, aged 70 years or more and current smokers. In addition, participants with granular stomatitis displayed increased amounts of plaque. CONCLUSION: The data presented here confirms the central role of denture biofilm accumulation in denture stomatitis. It also indicates that smoking may increase susceptibility to infection in its most severe form. PMID- 19545320 TI - Domestic use of a disclosing solution for denture hygiene: a randomised trial. AB - OBJECTIVES: The purpose of this study was to investigate the effect of the domestic use of a disclosing agent for denture hygiene. MATERIALS AND METHODS: Completely edentulous participants wearing maxillary dentures were randomly assigned to one of the three intervention groups: (1) Follow-up only (control; n = 12); (2) Oral and denture hygiene instructions (n = 10); (3) Instructions associated with the home use of a disclosing agent (1% neutral red; n = 10). Biofilm coverage area (%) over internal and external surfaces of the maxillary denture was assessed at baseline and after 14 and 90 days. Data were evaluated by generalised estimating equations based on score tests (alpha = 0.05). RESULTS: The participants presented low changes for areas of biofilm coverage (14 days (%): internal: GI = 1.4 +/- 0.9; GII = 1.5 +/- 1.3; GIII = -0.4 +/- 0.9; external: GI = 1.4 +/- 1.5; GII = 1.5 +/- 1.4; GIII = -0.4 +/- 0.9; 90 days (%): internal: GI = 2.0 +/- 0.9; GII = 2.2 +/- 1.4; GIII = 0.3 +/- 1.0; external: GI = 2.1 +/- 1.4; GII = 2.2 +/- 1.5; GIII = 0.3 +/- 0.9). Changes were similar for the three groups (p = 0.293) and were not influenced by the test time (p = 0.218). CONCLUSION: It can be concluded that the home use of a disclosing agent for denture hygiene does not improve the removal of the biofilm, particularly for patients with adequate oral hygiene habits. PMID- 19545321 TI - Why do some people lose teeth across their lifespan whereas others retain a functional dentition into very old age? AB - OBJECTIVES: To analyse the importance of caries, periodontitis, and medical and psychosocial factors for risk of becoming edentulous across their lifespan and to examine factors critical for retaining functional dentition into very old age. METHODS: From the longitudinal population-based Octogenarian Twin study which analysed psychosocial and health variables, 357 individuals aged 82 + in 1995 1998 were collected. Information about number of teeth, decayed and filled surfaces percentage and periodontal disease experience were drawn from dental records. Reasons for and time of edentulousness were recorded. RESULTS: Outcome varied - depending on perspective and factors for losing or retaining teeth. Significant factors for losing teeth varied over the lifespan. Losing teeth early in life was related to lower social class; in middle age, to lower education; and in old age, to poor lifestyle factors and low social class. Caries constituted the main reason for tooth loss (about 55%). This increased substantially in the >80 year age-group (75%). Maintaining a functional dentition into old age was significantly associated with non-smoking, more education, being married and good periodontal health. CONCLUSION: It is important to apply life-span and cohort perspectives to oral health and disease. In our sample of persons born before World War I, caries was the main reason for losing all teeth, with substantially increased prevalence by age. Lifestyle factors were significant for losing and for retaining teeth. Periodontal condition had a significant influence on the likelihood of retaining functional dentition, and also when taking psychosocial variables into account. PMID- 19545322 TI - Evaluation of three indices for biofilm accumulation on complete dentures. AB - OBJECTIVES: The objective of this study was to evaluate the accuracy and reproducibility of three complete denture biofilm indices (Prosthesis Hygiene Index; Jeganathan et al. Index; Budtz-Jorgensen Index) by means of a computerised comparison method. BACKGROUND: Clinical studies into denture hygiene have employed a large number of biofilm indices among their outcome variables. However, the knowledge about the validity of these indices is still scarce. MATERIALS AND METHODS: Sixty-two complete denture wearers were selected. The internal surfaces of the upper complete dentures were stained (5% erythrosine) and photographed. The slides were projected on paper, and the biofilm indices were applied over the photos by means of a scoring method. For the computerised method, the areas (total and biofilm-covered) were measured by dedicated software (Image Tool). In addition, to compare the results of the computerised method and Prosthetic Hygiene Index, a new scoring scale (including four and five graded) was introduced. For the Jeganathan et al. and Budtz-Jorgensen indices, the original scales were used. Values for each index were compared with the computerised method by the Friedman test. Their reproducibility was measured by means of weighed kappa. Significance for both tests was set at 0.05. RESULTS: The indices tested provided similar mean measures but they tended to overestimate biofilm coverage when compared with the computerised method (p < 0.001). Agreement between the Prosthesis Hygiene Index and the computerised method was not significant, regardless of the scale used. Jeghanathan et al. Index showed weak agreement, and consistent results were found for Budtz-Jorgensen Index (kappa = 0.19 and 0.39 respectively). CONCLUSION: Assessment of accuracy for the biofilm indices showed instrument bias that was similar among the tested methods. Weak inter-instrument reproducibility was found for the indices, except for the Budtz-Jorgensen Index. This should be the method of choice for clinical studies when more sophisticated approaches are not possible. PMID- 19545323 TI - Oral health care in long-term care facilities for elderly people in southern Brazil: a conceptual framework. AB - OBJECTIVE: To present a theoretical model for understanding oral health care for the elderly in the context of long-term care institutions (LTCI). METHODS: Open ended individual interviews were conducted with the elderly residing in LTCI, their carers, nursing technicians and nurses, directors of care, dental surgeons and managers of public health services. A grounded theory methodological approach was adopted for data collection and analysis. RESULTS: The emerging core category revealed a basic social process: 'Promoting oral health care for the elderly based on the context of LTCI'. This process was composed of two contradicting yet correlated aspects: the oral health care does not minimise the poor oral epidemiological condition, and at the same time, there was a continued improvement in the oral care expressed by better care practices. These aspects were related to the: attribution of meaning to oral health, social determination of oral health, the ageing process, interactions established in the oral health care practices, oral health care management in LTCI, inclusion of oral health care into the political-organisational dimension and possibility of conjecturing better oral health care practices. CONCLUSION: The core concept of 'Promotion of oral health care for elderly people based on the context of LTCI' is capable of explaining the variations in the structure and process of LTCI, as well as in helping to understand the meaning of the oral health care practices for the institutionalised elderly. PMID- 19545324 TI - Undergraduate teaching in gerodontology in Leipzig and Zurich--a comparison of different approaches. AB - OBJECTIVE: To evaluate undergraduate students' attitude towards the clinical components of the Leipzig (LPEG) and Zurich (ZPEG) Programmes of Education in Gerodontology. BACKGROUND: Undergraduate student education is the seedbed for conscientious professionals. Extramural clinical education contributes to the formation of positive attitudes. Students in Zurich participate in three clinical activities (in-house gerodontology clinic, extramural acute geriatrics ward, mobile dental service), in Leipzig they visit a long-term care facility on six occasions within 4 years. METHODS: A structured questionnaire with 10 items was administered to students in Leipzig [n = 34, 70.6% female, mean age 25.8 (SD 3.04) years] at the beginning and after completion of gerodontology training and to students in Zurich [n = 33, 48.5% female, mean age 27.0 (SD 3.28) years] on three occasions after clinical training. Students indicated the degree of their agreement with seven statements presented using a 5-point scale. A choice of responses which characterised the course was offered for assessment. RESULTS: Close collaboration with dental tutors, while self-treating patients in the mobile dental service (mobiDent) attracted the most positive responses. Ratings from students completing their training in Leipzig were less favourable than their initial responses. CONCLUSION: The lack of a dental service and Leipzig students' inability to offer treatment in the presence of disease was associated with frustrations. Practical training should go beyond dental examinations at a long-term care facility and include the opportunity for dental treatment. Personnel and equipment required for mobile treatment exceed resources available at most German dental schools. PMID- 19545325 TI - Non-surgical treatment of gingival overgrowth induced by nifedipine: a case report on an elderly patient. AB - Drug-induced gingival overgrowth (DIGO) is a significant problem for periodontologists and this side effect is frequently associated with three particular drugs: phenytoin, cyclosporin A and nifedipine. A case report of gingival overgrowth induced by nifedipine in an elderly patient treated with non surgical periodontal therapy is described. A 75-year-old male with generalised gingival overgrowth reported the problem of oral malodour and significant gingival bleeding. The medical history revealed a controlled hypertensive state and Cerebral Vascular Accident (CVA) 3 years prior to consultation. The diagnosis was gingival overgrowth associated with nifedipine, no other risk factors being identified. The patient had been taking nifedipine for 18 months, but after the consultation with the patient's doctor, nifedipine was suspended, as the hypertension was controlled. Treatment consisted of meticulous oral hygiene instruction, scaling, root surface instrumentation and prophylaxis. Six months after the first intervention, clinical parameters revealed a significant improvement with a considerable reduction in gingival overgrowth, demonstrating the effect of non-surgical periodontal therapy in severe cases of gingival overgrowth. Non-surgical treatment of DIGO is a far less invasive technique than surgical approaches and has demonstrated an impressively positive treatment response. It should therefore be considered as a first treatment option for DIGO. PMID- 19545326 TI - Relining of prosthesis with auto-polymerizing hard denture reline resins: effect of post-polymerization treatment on flexural strength. AB - BACKGROUND: It has been suggested that microwave irradiation and prosthesis immersion in hot water after its polymerization may improve mechanical and viscoelastic properties of acrylic resins. PURPOSE: This study was proposed to verify the influence of microwave post-polymerization (PP) treatment over the flexural strength of thermo-polymerizing acrylic resin specimens (QC-20) relined or not with two different composition hard chairside auto-polymerizing reliners [Kooliner (K) and New Truliner (NT)]. MATERIALS AND METHODS: For this study, 50 specimens of 64 x 10 x 3.3 mm were polymerized and distributed into five groups. G1 (control) specimens without relining and PP; G2 specimens relined with K, without PP; G3 specimens relined with NT, without PP; G4 specimens relined with K, with PP (microwave irradiation with 650 W for 5 min); G5 specimens relined with NT, with PP. Tests were performed on a universal testing machine Instron 4411 with compression speed of 5 mm/min. RESULTS: Specimens of K without PP did not show statistically different results (p < 0.05) when compared with control. However, when submitted to PP these specimens showed a significant increase in flexural strength. Specimens of NT showed the lowest flexural strength of all groups, with or without PP when compared with control and K groups. CONCLUSION: Microwave PP (650 W for 5 min) proved to be an effective method of improving the flexural strength of K relined prosthesis. However, it did not seem to affect NT specimens. PMID- 19545327 TI - Trends in NHS primary dental care for older people in England: implications for the future. AB - OBJECTIVE: To examine trends over time in the National Health Service (NHS) dental service provision for older people in England and consider the implications for future care. BACKGROUND: The number and proportion of older people in the population nationally are increasing and their oral health needs are changing as more people retain teeth into older age. MATERIALS AND METHODS: Descriptive analysis of routinely collected activity data for primary dental care in England was undertaken over the 6-year period, 1999/2000-2004/05. Data were cleaned, grouped into age bands for analysis. Activity rates per head of population were calculated for key aspects of care. Population forecasts and national survey data were used to inform future predictions of care required. RESULTS: There was an increase in absolute terms of total items (14.4%) and items per capita (10.7%) between 1999/2000 and 2004/05, with a peak of 15.7 million items claimed in 2003/04. Over the 6-year period, increases per capita were seen for dental examinations (11.3%), periodontal treatments (8.9%), fillings (1.9%), crowns (14.3%) and extractions (4.9%) amongst older people as a group. In contrast, there was a marked decrease in the episodes of care involving domiciliary care (-46.4%) and upper complete dentures (-32.9%) per capita over this period, items that were most commonly provided for the oldest age-band (85 years and over). Apart from complete dentures, the demand for care is predicted to increase. CONCLUSION: The volume of primary dental care provided for older people has been increasing at a rate which exceeds population growth, but remains low. Planners and commissioners should urgently review the provision of dental care of older people to ensure that their oral health needs are being met particularly in relation to dentures and domiciliary care. PMID- 19545328 TI - The spectrum of electrocardiographic manifestations of Brugada syndrome. PMID- 19545329 TI - Brugada-like changes in the peripheral leads during diagnostic ajmaline test in patients with suspected Brugada syndrome. AB - BACKGROUND: Although cases of Brugada-type electrocardiographic (ECG) pattern in peripheral (limb) leads have been reported ("atypical" Brugada syndrome [BS]), their incidence in patients investigated for BS is unknown. METHODS: We retrospectively analyzed an ECG database collected during ajmaline test in 143 patients (89 men) with suspected BS. In 42 patients, 12-lead ECGs were recorded, whereas in 101 patients, leads V1-V3 from the third intercostal space were also recorded. The presence of types 1, 2, and 3 Brugada pattern in each limb and precordial lead was noted and the PR, QRS, and QTc intervals were calculated. RESULTS: There were 114 (79.7%) negative and 29 (20.3%) positive tests. Type 1 pattern developed in >or=1 limb lead in six patients (4.2%) (3/29 with positive tests, 10.3%); all of them were male, symptomatic, and/or with family history of BS or sudden cardiac death. Their pre- and posttest QRS were significantly longer compared with the rest with positive (n = 26) or negative (n = 111) test (pretest: 129 +/- 31 ms vs 101 +/- 11 ms and 97 +/- 12 ms, P < 0.001; posttest: 175 +/- 44 ms vs 134 +/- 14 ms and 131 +/- 19 ms, P < 0.001). The posttest QTc was longer in patients with peripheral changes compared with the rest (507 +/- 47 ms vs 453 +/- 22 ms and 447 +/- 24 ms, P < 0.001). The pretest QTc and pre- and posttest heart rate and PR intervals were not significantly different between the three groups. CONCLUSIONS: Type 1 Brugada pattern in the peripheral leads was observed in 4.2% of patients during ajmaline test (10.3% of positive tests) and was associated with longer QRS and greater QTc prolongation compared with the rest of the patients. PMID- 19545330 TI - Prevalence of a Brugada pattern electrocardiogram in an urban population in the United States. AB - OBJECTIVE: To determine the prevalence of a Brugada-type pattern on routine electrocardiogram (ECG) in an urban population served by a tertiary medical center in the United States. METHODS: The investigators reviewed the ECG database at the Montefiore Medical Center, a tertiary teaching center in the Bronx, New York, over a 10-year period. During this time, 653,006 ECG records in 162,590 patients were identified. The database was queried by applying standard diagnostic criteria in an attempt to identify records with apparent conduction delay and ST abnormality in leads V1-V3. Additional diagnostic criteria were then applied to identify records in an attempt to mimic Brugada-like changes. A cardiac electrophysiologist reviewed records meeting these criteria to confirm the presence of a Brugada-type pattern. RESULTS: In total, 16,067 patients (9.8%) were identified as having ECGs with right bundle branch block, incomplete right bundle branch block, or RSR' in leads V1 and V2. After applying additional diagnostic criteria evaluating ST segment shift, 456 patients were identified as having a pattern potentially consistent with a Brugada-type ECG. The presence of a Brugada-type pattern was confirmed by physician overread in 20 patients (0.012%). CONCLUSION: The Brugada-type ECG pattern is infrequently seen in a large ethnically diverse urban US population. Further evaluation should be considered when this pattern is seen on routine ECG. PMID- 19545331 TI - Esophageal damage during catheter ablation of atrial fibrillation: is cryo safer than RF? PMID- 19545332 TI - Esophageal contour changes during cryoablation of atrial fibrillation. AB - BACKGROUND: We have observed contour changes of the barium-filled esophagus during atrial fibrillation (AF) ablation with cryo-energy delivered in direct proximity to the esophagus. OBJECTIVE: To evaluate the frequency, location, and severity of esophageal contour changes during cryo-energy application close to the esophagus. METHODS: We retrospectively analyzed cine-fluoroscopic images acquired during hybrid cryo-radiofrequency AF ablation in 100 consecutive patients with cryo-energy delivered only in direct proximity to the esophagus. RESULTS: Esophageal contour changes were observed in 28 (32%) of 89 patients (and 74 [6.2%] of 1,191 of all cryo applications). They were more frequent in the left common pulmonary vein (PV) (50%) and less so in the right common PV and the upper PVs (4-5%). The distance of the ablation catheter from the endoesophageal contour prior to cryo-energy applications associated with contour changes was 1.8 +/- 1.5 mm, which increased to 4.1 +/- 1.6 mm at the time of peak contour change (P < 0.001). The esophageal contour deformation was 2.3 +/- 0.9 mm. There were no apparent complications related to cryo-energy application for 3-4 minutes, even if associated with contour changes. CONCLUSION: Esophageal contour changes were observed in >6% of cryo applications in direct proximity to the esophagus (32% of patients) and were most frequent in the posterior aspect of the left common and right lower PV ostium when cryo-energy was delivered at a distance of 1.5 mV as "high-voltage zone." Successful ablation sites were electroanatomically classified into each voltage zone. RESULTS: Successful ablation was acquired in 63 patients and 71 RVOT arrhythmias (63/72 patients: 87.5%, 71/82 RVOT arrhythmias: 86.5%). In the successful group, three arrhythmias (4.2%) were classified in the low-voltage zone, 63 arrhythmias (88.7%) in the transitional-voltage zone, and five arrhythmias (7.0%) in the high-voltage zone. CONCLUSIONS: This study indicates that the vast majority of successful ablation sites for idiopathic RVOT arrhythmias are located in the transitional-voltage zone. Mapping of the transitional-voltage zone may be an important landmark of RFCA for RVOT arrhythmia. PMID- 19545335 TI - Assessment of clockwise cavotricuspid isthmus block based on conduction times during transient entrainment: a prospective study. AB - BACKGROUND: In typical counterclockwise atrial flutter (AFL), the route of impulse propagation to anteroinferior right atrium (AIRA) during transient entrainment (TE) from the coronary sinus (CS) is expected to be similar to that during pacing from the same CS site during sinus rhythm (SR) when cavotricuspid isthmus (CTI) block has occurred. This could be used to identify CTI block during ablation procedures. METHODS: Thirty-six patients with AFL (cycle length [CL], 240 +/- 25 ms) underwent CTI ablation during AFL. CS pacing was performed at a CL of 20 ms less than AFL CL before ablation (n = 36), and at several CL during SR with conduction through the CTI (n = 21) and after CTI block (n = 36). RESULTS: TE with orthodromic activation of AIRA occurred in all 36 patients. Conduction time from CS to AIRA during TE (T-entr, 199 +/- 29 ms) was significantly longer than during pacing in SR (T-CTI) at the same rate not only with CTI conduction (T CTI-C, 135 +/- 24 ms, P < 0.001), but also with CTI block (T-CTI-B, 186 +/- 24 ms, P < 0.01). T-entr did not correlate with T-CTI-C, but there was an excellent correlation between T-entr and T-CTI-B (r = 0.874, P < 0.001). A "TE index" that corrected T-CTI for individual T-entr identified CTI block with 97% sensitivity and 91% specificity. T-CTI at low rates differed from T-CTI at high rates but correlated significantly with them. CONCLUSION: Comparison of conduction times during TE from the CS and during pacing from the same site and rate in SR can help to establish whether clockwise CTI block has been achieved in patients with typical AFL. PMID- 19545336 TI - The influence of left ventricle diastolic function on natriuretic peptides levels in patients with atrial fibrillation. AB - BACKGROUND: The diagnosis of the impaired left ventricle (LV) diastolic function during atrial fibrillation (AF) using traditional methods is very difficult. Natriuretic peptides seem to be useful for assessment of diastolic function in patients with AF. AIM: To evaluate the influence of LV diastolic dysfunction on natriuretic peptides concentrations and to assess the diagnostic value of atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) in patients with AF and impaired LV diastolic function. METHODS: The study included 42 patients (23 males, 19 females), aged 58.6 +/- 8.2 years with nonvalvular persistent AF with preserved LV systolic function who were converted into sinus rhythm by DC cardioversion (CV) and maintained sinus rhythm for at least 30 days. Echocardiography (ECG), ANP, and BNP level measurements were taken at baseline 24 hours before CV and 24 hours and 30 days after CV. On the 30th day following CV in patients with sinus rhythm, Doppler ECG was performed to assess LV diastolic function. RESULTS: Thirty days after CV, normal LV diastolic function in 15 patients and impaired diastolic function in 27 patients was diagnosed: 20 with impaired LV relaxation and seven with impaired LV compliance. During AF and 24 hours, and 30 days after sinus rhythm restoration, significantly higher ANP and BNP levels were observed in patients with LV diastolic dysfunction as compared to the subgroup with normal LV diastolic function. The average values of ANP during AF in patients with normal and impaired diastolic function were 167.3 +/- 70.1 pg/mL and 298.7 +/- 83.6 pg/mL, respectively (P < 0.001), and the average values of BNP in the above mentioned subgroups were 49.5 +/- 14.7 pg/mL and 145.6 +/- 49.6 pg/mL respectively (P < 0.001). While comparing the diagnostic value of both natriuretic peptides it was noted that BNP was a more specific and sensitive marker of impaired LV diastolic function. ANP value >220.7 pg/mL measured during AF identified patients with impaired LV diastolic function with 85% sensitivity and 90% specificity. BNP value >74.7 pg/mL proved 95% sensitive and 100% specific in the diagnosing of such a group. CONCLUSIONS: The increase of ANP/BNP concentration in patients with AF results not only from the presence of AF, but also reflects the impaired LV diastolic function. Natriuretic peptides, especially BNP, may be useful in diagnosing LV diastolic dysfunction in patients with AF. PMID- 19545337 TI - The role of electrophysiologic studies in dilated cardiomyopathy: definitive results from the DEFINITE trial. PMID- 19545338 TI - Ventricular arrhythmia inducibility predicts subsequent ICD activation in nonischemic cardiomyopathy patients: a DEFINITE substudy. AB - OBJECTIVES: We evaluated whether electrophysiologic (EP) inducibility predicts the subsequent occurrence of spontaneous ventricular tachycardia (VT) or ventricular fibrillation (VF) in the Defibrillators in Nonischemic Cardiomyopathy Treatment Evaluation (DEFINITE) trial. BACKGROUND: Inducibility of ventricular arrhythmias has been widely used as a risk marker to select implantable cardioverter defibrillator (ICD) candidates, but is believed not to be predictive in nonischemic cardiomyopathy patients. METHODS: In DEFINITE, patients randomized to the ICD arm, but not the conventional arm, underwent noninvasive EP testing via the ICD shortly after ICD implantation using up to three extrastimuli at three cycle lengths plus burst pacing. Inducibility was defined as monomorphic or polymorphic VT or VF lasting 15 seconds. Patients were followed for a median of 29 +/- 14 months (interquartile range = 2-41). An independent committee, blinded to inducibility status, characterized the rhythm triggering ICD shocks. RESULTS: Inducibility, found in 29 of 204 patients (VT in 13, VF in 16), was associated with diabetes (41.4% vs 20.6%, P = 0.014) and a slightly higher ejection fraction (23.2 +/- 5.9 vs 20.5 +/- 5.7, P = 0.021). In follow-up, 34.5% of the inducible group (10 of 29) experienced ICD therapy for VT or VF or arrhythmic death versus 12.0% (21 of 175) noninducible patients (hazard ratio = 2.60, P = 0.014). CONCLUSIONS: In DEFINITE patients, inducibility of either VT or VF was associated with an increased likelihood of subsequent ICD therapy for VT or VF, and should be one factor considered in risk stratifying nonischemic cardiomyopathy patients. PMID- 19545339 TI - Acute evaluation of transthoracic impedance vectors using ICD leads. AB - BACKGROUND: Minute ventilation (MV) has been proven to be very useful in rate responsive pacing. The aim of this study was to evaluate the feasibility of using implantable cardioverter-defibrillator (ICD) leads as part of the MV detection system. METHODS: At implant in 10 patients, the transthoracic impedance was measured from tripolar ICD, tetrapolar ICD, and atrial lead vectors during normal, deep, and shallow voluntary respiration. MV and respiration rate (RespR) were simultaneously measured through a facemask with a pneumotachometer (Korr), and the correlations with impedance-based measurements were calculated. Air sensitivity was the change in impedance per change in respiratory tidal volume, ohms (Omega)/liter (L), and the signal-to-noise ratio (SNR) was the ratio of the respiratory and cardiac contraction components. RESULTS: The air sensitivity and SNR in tripolar ICD vector were 2.70 +/- 2.73 ohm/L and 2.19 +/- 1.31, respectively, and were not different from tetrapolar. The difference in RespR between tripolar ICD and Korr was 0.2 +/- 1.91 breaths/minute. The regressed correlation coefficient between impedance MV and Korr MV was 0.86 +/- 0.07 in tripolar ICD. CONCLUSIONS: The air sensitivity and SNR in tripolar and tetrapolar ICD lead vectors did not differ significantly and were in the range of the values in pacemaker leads currently used as MV sensors. The good correlations between impedance-based and Korr-based RespR and MV measurements imply that ICD leads may be used in MV sensor systems. PMID- 19545340 TI - Ectopy in patients with permanent pacemakers and implantable cardioverter defibrillators undergoing an MRI scan. AB - BACKGROUND: Recent series suggest that magnetic resonance imaging (MRI) scanning can be performed safely in select patients with pacemakers or implantable cardioverter-defibrillators (ICDs). Limited data have been reported on ectopy during MRI scans in patients with pacemakers or ICDs. This study evaluated increased ectopy observed in patients with permanent pacemakers or ICDs undergoing MRI scanning of any landmark without peak specific absorption rate (SAR) limit. METHODS: Fifty-two patients with a total of 119 leads underwent a total of 59 MRI scans of any landmark using usual protocols with standard peak SAR settings for the scan. No patient was pacemaker dependent. All devices were programmed to single-chamber demand mode (VVI) or dual-chamber demand mode (DDI) with a lower rate of 40 bpm.Both telemetry and pulse oximetry plethysmographic waveform were observed continuously throughout the scans for ectopy. RESULTS: Increased ectopy was observed during seven scans. The ectopy in four scans was ventricular and had fixed coupling intervals of 1,500 and 3,000, and was likely due to device noise rejection behavior. The etiology of ectopy observed during the other three scans could not be determined. Ectopy could not be predicted by peak SAR, scan time duration, or landmark. No significant changes in pacing thresholds were seen postscan. CONCLUSIONS: The current series suggests that a minority of patients with implanted pacemakers may have MRI-related ectopy. A significant proportion of this ectopy may arise from normal device behavior within the MRI environment. PMID- 19545341 TI - Permanent atrial pacing lead implant route after Fontan operation. AB - BACKGROUND: Atrial pacing is indicated for sinus node dysfunction (SND) after Fontan surgery; preferred lead implantation technique is debated. We compare outcomes of transvenous (TV) and epicardial (Epi) atrial lead implants in this population. METHODS: Retrospective review of Fontan patients undergoing atrial lead implant between 1992 and 2007. Demographics, lead performance data, and outcomes were analyzed. RESULTS: 78 patients had 90 leads implanted: 25 via TV route and 65 via Epi route. Median follow-up was 1.6 years (TV) and 3.6 years (Epi). TV leads were implanted in older patients (23.1 vs 9.3 years, P < 0.001) and at longer intervals after Fontan (15.2 vs 4.9 years, P < 0.001). Pacing indication for most TV leads was SND, while Epi leads were also indicated for atrioventricular block. Acute complication rates were similar (8% TV vs 19% Epi, P = 0.23), but median hospital stay was shorter for TV (2 vs 5 days, P = 0.03). Thrombus was observed in five patients (two in TV; three in Epi), but no thromboembolic events were observed. Mean lead survival was similar (TV 9.9 vs Epi 7.8 years, P = NS). Energy threshold was lower at implant for TV leads (0.9 vs 2.2 microJ, P = 0.049), but similar at follow-up (1.2 vs 2.6 microJ, P = 0.35). Atrial sensing was unchanged over time for TV (2.2 to 2.1 mV, P = NS), but decreased for Epi (3.3 to 2.5 mV, P = 0.02). CONCLUSIONS: Compared to epicardial leads, transvenous atrial pacing leads may be placed in Fontan patients with lower procedural morbidity and equivalent expectation of lead performance and longevity. PMID- 19545342 TI - On an improved T-wave alternans index: a hypothesis. PMID- 19545343 TI - The road not taken. PMID- 19545344 TI - Implantable cardioverter-defibrillator discharges despite successful termination of ventricular tachycardia by antitachycardia pacing. PMID- 19545345 TI - BNP/NT-proBNP and cardiac pacing: a review. AB - Natriuretic peptide (NP) levels (B-type natriuretic peptide [BNP] and N-terminal proBNP) are now widely used in clinical practice and cardiovascular research all over the world and have been incorporated into many cardiovascular guidelines for heart failure (HF). The roles of NP levels are evolving rapidly not only in diagnosis, therapy monitoring, and risk stratification of HF, but also in differential diagnosis of acute dyspnea, predicting death and rehospitalization in HF patients. NP assays have been applied in permanent cardiac pacing in recent years, whereas it is still not well known how NP levels change and whether NP levels can predict HF in permanent cardiac pacing. Therefore, this article reviews the role of NP levels in permanent cardiac pacing, mainly including NP changes in different cardiac pacing modes and cardiac resynchronization therapy. PMID- 19545346 TI - Troponin levels after cardiac electrophysiology procedures: review of the literature. AB - Cardiac-specific Troponins (cTn) I and T have become markers of choice for myocardial injury. We reviewed the literature in order to understand the expected postprocedure cTn rise after electrophysiology procedures. A total of 34 studies and 1,608 patients were included. After external monophasic cardioversion, seven of 442 patients (1.6%) had minimal increase in cTnI (0.1-0.9 ng/mL), and only one of 368 (0.3%) had increase in cTnT (0.11 ng/mL). After internal cardioversion, 17 of 105 (16%) had increase in cTnI (0.7-2.4 ng/mL) but only three (2.9%) were above the cutoff for myocardial infarction (MI) (1.5 ng/mL). During implantable cardioverter-defibrillator (ICD) installation with a mean of 2-7 ICD shocks, 12 of 74 (16%) patients had cTnI >or=1.5 ng/mL (range 1.7-5.5 ng/mL) and 20 of 64 (32%) had cTnT >or=0.1 ng/mL (range 0.26-6.46 ng/mL) considered in the range of clinical MI. Radio frequency ablation (RFA) (n = 496) resulted in significant cTn elevation in 25-100% of patients with ventricular > atrial and linear > focal lesions. Average postprocedure peak cTnI ranged from 0.13 to 6 ng/mL (median: 2.36 ng/mL, max: 15 ng/mL) and cTnT 0.2 to 2.41 ng/mL (median: 0.44 ng/mL, max: 9 ng/mL). Early cTn peak at 2-8 hours was noted after RFA. External cardioversion should not cause a significant increase in cTn; RFA and ICD implantation with shocks often result in an increase in cTn. Interpretation of these markers can be difficult if acute coronary syndrome is suspected in the postprocedure period. PMID- 19545347 TI - Pilsicainide-induced ST segment elevation and ST segment depression in two patients with variant forms of Brugada-type electrocardiographic abnormalities. AB - In two patients with variant forms of Brugada electrocardiographic abnormalities, ST segment elevation, and reciprocal ST segment depression developed during intravenous administration of pilsicainide. In one patient, pilsicainide accentuated the ST segment elevation in leads I, aV(L), and V(1)-V(3) and caused ST segment depression in leads II, III, and aV(F). Coronary angiograms at the time of ST segment elevation were normal. In the other patient, pilsicainide accentuated the coved-type ST segment elevation in leads II, III, and aV(F) and caused ST segment depression in leads I, aV(L), and V(2)-V(5). Frequent premature ventricular complexes (PVCs) with two different left bundle branch block patterns developed during ST segment elevation. Intravenous isoproterenol returned the ST segment to baseline in both patients and suppressed the PVCs in the second patient. We hypothesize that a wide area of epicardial myocardium with large I(to) current might explain the reciprocal ST segment depression observed at the time of accentuated ST segment elevation. PMID- 19545348 TI - A case of reversible ventricular tachycardia and complete atrioventricular block associated with primary cardiac B-cell lymphoma. AB - We report a long-term survival case of primary cardiac lymphoma with reversible ventricular tachycardia (VT) and complete atrioventricular block (C-AVB). A 65 year-old man with VT was treated by oral amiodarone administration. Later, a dual chamber pacemaker was implanted because of C-AVB. Then, he was readmitted, as he complained of fever and chest pain. Echocardiography showed an enlarged cardiac mass and thus an open-chest biopsy was performed. He was then diagnosed with primary cardiac lymphoma. The chemotherapy and radiotherapy resulted in the disappearance of the mass. Complete remission has been maintained for 8 years after the therapy, and no VT or C-AVB has been detected. PMID- 19545349 TI - Giant left atrial thrombus associated with ablation for atrial tachycardia. AB - We report a very rare case of giant left atrial thrombus (size: 7.2 x 4.5 mm(2)) associated with radiofrequency catheter ablation for atrial tachycardia in a 72 year-old man. After 4 weeks of anticoagulation with warfarin, a repeat echocardiogram demonstrated partial resolution of the thrombus (size: 4.5 x 2.6 mm(2)) without systemic embolization. PMID- 19545350 TI - Use of a wireless implantable cardioverter defibrillator in a patient with a preexisting neurostimulator. AB - We report the case of a 54-year-old man with a previously implanted neurostimulator who presented with palpitations and was found to have sustained ventricular tachycardia on electrophysiologic study. A Medtronic wireless implantable cardioverter defibrillator (ICD; Medtronic Inc., Minneapolis, MN, USA) with home monitoring (HM) was successfully implanted. Interaction testing during implantation, follow-up, and HM showed that there was no device-device interaction. PMID- 19545351 TI - Catheter ablation of premature ventricular contractions arising from the mitral annulus after mitral valvoplasty. AB - A 57-year-old man undergoing mitral valvoplasty underwent catheter ablation of symptomatic premature ventricular contractions (PVCs) with a right bundle branch block and right inferior axis QRS morphology. Left ventriculography revealed a normal left ventricular function and visualized the anatomical relationships between the left ventricular outflow tract and the mitral annuloplasty ring. Electroanatomic mapping during the PVCs revealed a centrifugal activation pattern arising from the mitral annulus, and the PVCs were likely to be idiopathic. Successful radiofrequency ablation was achieved at the site close to the antero paraseptal end of the mitral annuloplasty ring, which was located adjacent to the fibrous trigone. PMID- 19545352 TI - Successful balloon dilatation of the valve of Vieussens for left ventricular lead placement. AB - Successful balloon dilatation of an obstructive valve of Vieussens for left ventricular lead placement is described in a case with severe left ventricular systolic dysfunction. PMID- 19545353 TI - Microbial modification of host long-distance dispersal capacity. AB - BACKGROUND: Dispersal plays a key role in shaping biological and ecological processes such as the distribution of spatially-structured populations or the pace and scale of invasion. Here we have studied the relationship between long distance dispersal behaviour of a pest-controlling money spider, Erigone atra, and the distribution of maternally acquired endosymbionts within the wider meta population. This spider persists in heterogeneous environments because of its ability to recolonise areas through active long-distance airborne dispersal using silk as a sail, in a process termed 'ballooning'. RESULTS: We show that there is spatial heterogeneity in the prevalence of two maternally acquired endosymbiont infections within the wider E. atra meta-population and we demonstrate through several independent approaches a link between the presence of one of these endosymbionts, Rickettsia, and the tendency for long-distance movement. CONCLUSION: This novel finding that particular endosymbionts can influence host dispersal is of broad importance given the extremely widespread occurrence of similar bacteria within arthropod communities. A bacterial phenotype that limits dispersal has the potential not only to reduce gene flow and thus contribute to degrees of reproductive isolation within species, but also to influence species distribution and thus overall community composition. PMID- 19545354 TI - Genomic characterization of the human mitochondrial tumor suppressor gene 1 (MTUS1): 5' cloning and preliminary analysis of the multiple gene promoters. AB - BACKGROUND: Mitochondrial tumor suppressor gene 1 (MTUS1) has been recently identified as a candidate tumor suppressor gene which resides in a genomic region (8p22) that shows frequent loss of heterozygosity (LOH) in several tumor types. It has been suggested that multiple gene promoters and alternative splicing lead to the expression of 5 known MTUS1 transcript variants. FINDINGS: Here, we characterized the 5' untranslated regions of the different transcript variants. We also cloned and functionally tested the alternatively utilized gene promoters that contribute to the production of different MTUS1 transcript variants. CONCLUSION: Our results confirmed the early hypothesis that the transcript variants of MTUS1 gene are driven by multiple gene promoters. PMID- 19545355 TI - Hospital-based surveillance of enteric parasites in Kolkata. AB - BACKGROUND: Diarrhoea is the second leading cause of illness and death in developing countries and the second commonest cause of death due to infectious diseases among children under five in such countries. Parasites, as well as bacterial and viral pathogens, are important causes of diarrhoea. However, parasitic infections are sometimes overlooked, leading after a period of time to an uncertain aetiology. In this paper we report the prevalence of Giardia lamblia, Entamoeba histolytica and Cryptosporidium sp. in and around Kolkata. FINDINGS: A hospital-based laboratory surveillance study was conducted among the patients admitted between November 2007 and October 2008 to the Infectious Diseases (ID) Hospital (Population = 1103) with diarrhoeal complaints. Of the 1103 samples collected, 147 were positive for Giardia lamblia, 84 for Cryptosporidium sp. and 51 for Entamoeba histolytica. For all these parasites there was a high rate of mixed infection with common enteric viruses and bacteria such as Rotavirus, Vibrio cholerae and Shigella sp. There were also cases of co infection with all other diarrheogenic pathogens. The age group >or= 5 years had the highest prevalence of parasites whereas the age group >5 - 10 years was predominantly infected with Giardia lamblia (p =< 0.001; Odds ratio (OR) = 3.937; 95% Confidence interval (CI) = 1.862 - 8.326) and with all parasites (p = 0.040; OR = 2.043; 95% CI = 1.033 - 4.039). The age group >10 - 20 years could also be considered at risk for G. lamblia (p = 0.009; OR = 2.231; 95% CI = 1.223 - 4.067). Month-wise occurrence data showed an endemic presence of G. lamblia whereas Cryptosporidium sp. and E. histolytica occurred sporadically. The GIS study revealed that parasites were more prevalent in areas such as Tangra, Tiljala and Rajarhat, which are mainly slum areas. Because most of the population surveyed was in the lower income group, consumption of contaminated water and food could be the major underlying cause of parasitic infestations. CONCLUSION: This study provides important information on the occurrence and distribution of three important intestinal parasites and indicates their diarrheogenic capacity in Kolkata and surrounding areas. PMID- 19545357 TI - Health beliefs about bottled water: a qualitative study. AB - BACKGROUND: There has been a consistent rise in bottled water consumption over the last decade. Little is known about the health beliefs held by the general public about bottled water as this issue is not addressed by the existing quantitative literature. The purpose of this study was to improve understanding of the public's health beliefs concerning bottled mineral water, and the extent to which these beliefs and other views they hold, influence drinking habits. METHODS: A qualitative study using semi-structured interviews, with 23 users of the Munrow Sports Centre on the University of Birmingham campus. RESULTS: Health beliefs about bottled water could be classified as general or specific beliefs. Most participants believed that bottled water conferred general health benefits but were unsure as to the nature of these. In terms of specific health beliefs, the idea that the minerals in bottled water conferred a health benefit was the most commonly cited. There were concerns over links between the plastic bottle itself and cancer. Participants believed that bottled water has a detrimental effect on the environment. Convenience, cost and taste were influential factors when making decisions as to whether to buy bottled water; health beliefs were unimportant motivating factors. CONCLUSION: The majority of participants believed that bottled water has some health benefits. However, these beliefs played a minor role in determining bottled water consumption and are unlikely to be helpful in explaining recent trends in bottled water consumption if generalised to the UK population. The health beliefs elicited were supported by scientific evidence to varying extents. Most participants did not feel that bottled water conferred significant, if any, health benefits over tap water. PMID- 19545356 TI - Direct cord implantation in brachial plexus avulsions: revised technique using a single stage combined anterior (first) posterior (second) approach and end-to side side-to-side grafting neurorrhaphy. AB - BACKGROUND: The superiority of a single stage combined anterior (first) posterior (second) approach and end-to-side side-to-side grafting neurorrhaphy in direct cord implantation was investigated as to providing adequate exposure to both the cervical cord and the brachial plexus, as to causing less tissue damage and as to being more extensible than current surgical approaches. METHODS: The front and back of the neck, the front and back of the chest up to the midline and the whole affected upper limb were sterilized while the patient was in the lateral position; the patient was next turned into the supine position, the plexus explored anteriorly and the grafts were placed; the patient was then turned again into the lateral position, and a posterior cervical laminectomy was done. The grafts were retrieved posteriorly and side grafted to the anterior cord. Using this approach, 5 patients suffering from complete traumatic brachial plexus palsy, 4 adults and 1 obstetric case were operated upon and followed up for 2 years. 2 were C5,6 ruptures and C7,8T1 avulsions. 3 were C5,6,7,8T1 avulsions. C5,6 ruptures were grafted and all avulsions were cord implanted. RESULTS: Surgery in complete avulsions led to Grade 4 improvement in shoulder abduction/flexion and elbow flexion. Cocontractions occurred between the lateral deltoid and biceps on active shoulder abduction. No cocontractions occurred after surgery in C5,6 ruptures and C7,8T1 avulsions, muscle power improvement extended into the forearm and hand; pain disappeared. LIMITATIONS INCLUDE: spontaneous recovery despite MRI appearance of avulsions, fallacies in determining intraoperative avulsions (wrong diagnosis, wrong level); small sample size; no controls rule out superiority of this technique versus other direct cord reimplantation techniques or other neurotization procedures; intra- and interobserver variability in testing muscle power and cocontractions. CONCLUSION: Through providing proper exposure to the brachial plexus and to the cervical cord, the single stage combined anterior (first) and posterior (second) approach might stimulate brachial plexus surgeons to go more for direct cord implantation. In this study, it allowed for placing side grafts along an extensive donor recipient area by end-to-side, side-to-side grafting neurorrhaphy and thus improved results. LEVEL OF EVIDENCE: Level IV, prospective case series. PMID- 19545358 TI - Anthroposophic therapy for children with chronic disease: a two-year prospective cohort study in routine outpatient settings. AB - BACKGROUND: Many children with chronic disease use complementary therapies. Anthroposophic treatment for paediatric chronic disease is provided by physicians and differs from conventional treatment in the use of special therapies (art therapy, eurythmy movement exercises, rhythmical massage therapy) and special medications. We studied clinical outcomes in children with chronic diseases under anthroposophic treatment in routine outpatient settings. METHODS: In conjunction with a health benefit program, consecutive outpatients starting anthroposophic treatment for any chronic disease participated in a prospective cohort study. Main outcome was disease severity (Disease and Symptom Scores, physicians' and caregivers' assessment on numerical rating scales 0-10). Disease Score was documented after 0, 6, and 12 months, Symptom Score after 0, 3, 6, 12, 18, and 24 months. RESULTS: A total of 435 patients were included. Mean age was 8.2 years (standard deviation 3.3, range 1.0-16.9 years). Most common indications were mental disorders (46.2% of patients; primarily hyperkinetic, emotional, and developmental disorders), respiratory disorders (14.0%), and neurological disorders (5.7%). Median disease duration at baseline was 3.0 years (interquartile range 1.0-5.0 years). The anthroposophic treatment modalities used were medications (69.2% of patients), eurythmy therapy (54.7%), art therapy (11.3%), and rhythmical massage therapy (6.7%). Median number of eurythmy/art/massage therapy sessions was 12 (interquartile range 10-20), median therapy duration was 118 days (interquartile range 78-189 days).From baseline to six-month follow-up, Disease Score improved by average 3.00 points (95% confidence interval 2.76-3.24 points, p < 0.001) and Symptom Score improved by 2.41 points (95% confidence interval 2.16-2.66 points, p < 0.001). These improvements were maintained until the last follow-up. Symptom Score improved similarly in patients not using adjunctive non-anthroposophic therapies within the first six study months. CONCLUSION: Children under anthroposophic treatment had long-term improvement of chronic disease symptoms. Although the pre-post design of the present study does not allow for conclusions about comparative effectiveness, study findings suggest that anthroposophic therapies may play a beneficial role in the long-term care of children with chronic illness. PMID- 19545359 TI - Randomized controlled trial of the efficacy of aerobic exercise in reducing metabolic risk in healthy older people: The Hertfordshire Physical Activity Trial. AB - BACKGROUND: While there are compelling observational data confirming that individuals who exercise are healthier, the efficacy of aerobic exercise interventions to reduce metabolic risk and improve insulin sensitivity in older people has not been fully elucidated. Furthermore, while low birth weight has been shown to predict adverse health outcomes later in life, its influence on the response to aerobic exercise is unknown. Our primary objective is to assess the efficacy of a fully supervised twelve week aerobic exercise intervention in reducing clustered metabolic risk in healthy older adults. A secondary objective is to determine the influence of low birth weight on the response to exercise in this group. METHODS/DESIGN: We aim to recruit 100 participants born between 1931 1939, from the Hertfordshire Cohort Study and randomly assign them to no intervention or to 36 fully supervised one hour sessions on a cycle ergometer, over twelve weeks. Each participant will undergo detailed anthropometric and metabolic assessment pre- and post-intervention, including muscle biopsy, magnetic resonance imaging and spectroscopy, objective measurement of physical activity and sub-maximal fitness testing. DISCUSSION: Given the extensive phenotypic characterization, this study will provide valuable insights into the mechanisms underlying the beneficial effects of aerobic exercise as well as the efficacy, feasibility and safety of such interventions in this age group. TRIAL REGISTRATION: Current Controlled Trials: ISRCTN60986572. PMID- 19545360 TI - Tolerance of a standard intact protein formula versus a partially hydrolyzed formula in healthy, term infants. AB - BACKGROUND: Parents who perceive common infant behaviors as formula intolerance related often switch formulas without consulting a health professional. Up to one half of formula-fed infants experience a formula change during the first six months of life. METHODS: The objective of this study was to assess discontinuance due to study physician-assessed formula intolerance in healthy, term infants. Infants (335) were randomized to receive either a standard intact cow milk protein formula (INTACT) or a partially hydrolyzed cow milk protein formula (PH) in a 60 day non-inferiority trial. Discontinuance due to study physician-assessed formula intolerance was the primary outcome. Secondary outcomes included number of infants who discontinued for any reason, including parent-assessed. RESULTS: Formula intolerance between groups (INTACT, 12.3% vs. PH, 13.7%) was similar for infants who completed the study or discontinued due to study physician-assessed formula intolerance. Overall study discontinuance based on parent- vs. study physician-assessed intolerance for all infants (14.4 vs.11.1%) was significantly different (P = 0.001). CONCLUSION: This study demonstrated no difference in infant tolerance of intact vs. partially hydrolyzed cow milk protein formulas for healthy, term infants over a 60-day feeding trial, suggesting nonstandard partially hydrolyzed formulas are not necessary as a first-choice for healthy infants. Parents frequently perceived infant behavior as formula intolerance, paralleling previous reports of unnecessary formula changes. TRIAL REGISTRATION: clinicaltrials.gov: NCT00666120. PMID- 19545362 TI - Human rights of persons with mental illness in Indonesia: more than legislation is needed. AB - BACKGROUND: Although attention to human rights in Indonesia has been improving over the past decade, the human rights situation of persons with mental disorders is still far from satisfactory. The purpose of this paper is to examine the legal framework for protection of human rights of persons with mental disorder and the extent to which Indonesia's international obligations concerning the right to health are being met. METHODS: We examined the Indonesian constitution, Indonesian laws relevant to the right to health, the structure and operation of the National Human Rights Commission, and what is known about violations of the human rights of persons with mental illness from research and the media. RESULTS: The focus of the Indonesian Constitution on rights pre-dated the Universal Declaration, Indonesia has ratified relevant international covenants and domestic law provides an adequate legal framework for human rights protections. However, human rights abuses persist, are widespread, and go essentially unremarked and unchallenged. The National Human Rights Commission has only recently become engaged in the issue of protection of the rights of persons with mental illness. CONCLUSION: More than legislation is needed to protect the human rights of persons with mental illness. Improving the human rights situation for persons with mental illness in Indonesia will require action by governments at national, provincial and district levels, substantial increases in the level of investment in mental health services, coordinated action by mental health professionals and consumer and career organisations, and a central role for the National Human Rights Commission in protecting the rights of persons with mental illness. PMID- 19545361 TI - Cost-effectiveness of micafungin as an alternative to fluconazole empiric treatment of suspected ICU-acquired candidemia among patients with sepsis: a model simulation. AB - INTRODUCTION: Recent epidemiologic literature indicates that candidal species resistant to azoles are becoming more prevalent in the face of increasing incidence of hospitalizations with candidemia. Echinocandins, a new class of antifungal agents, are effective against resistant candidal species. As delaying appropriate antifungal coverage leads to increased mortality, we evaluated the cost-effectiveness of 100 mg daily empiric micafungin (MIC) vs. 400 mg daily fluconazole (FLU) for suspected intensive care unit-acquired candidemia (ICU-AC) among septic patients. METHODS: We designed a decision model with inputs from the literature in a hypothetical 1000-patient cohort with suspected ICU-AC treated empirically with either MIC or FLU or no treatment accompanied by a watchful waiting strategy. We examined the differences in the number of survivors, acquisition costs of antifungals, and lifetime costs among survivors in the cohort under each scenario, and calculated cost per quality adjusted life year (QALY). We conducted Monte Carlo simulations and sensitivity analyses to determine the stability of our estimates. RESULTS: In the base case analysis, assuming ICU-AC attributable mortality of 0.40 and a 52% relative risk reduction in mortality with appropriate timely therapy, compared with FLU (total deaths 31), treatment with MIC (total deaths 27) would result in four fewer deaths at an incremental cost/death averted of $61,446. Similarly, in reference case, incremental cost-effectiveness of MIC over FLU was $34,734 (95% confidence interval $26,312 to $49,209) per QALY. The estimates were most sensitive to the QALY adjustment factor and the risk of candidemia among septic patients. CONCLUSIONS: Given the increasing likelihood of azole resistance among candidal isolates, empiric treatment of ICU-AC with 100 mg daily MIC is a cost-effective alternative to FLU. PMID- 19545364 TI - Changes in physical health among participants in a multidisciplinary health programme for long-term unemployed persons. AB - BACKGROUND: The relationship between poor health and unemployment is well established. Health promotion among unemployed persons may improve their health. The aims of this study were to investigate characteristics of non-participants and drop-outs in a multidisciplinary health promotion programme for long-term unemployed persons with health complaints, to evaluate changes in physical health among participants, and to investigate determinants of improvement in physical health. METHODS: A longitudinal, non-controlled design was used. The programme consisted of two weekly exercise sessions and one weekly cognitive session during 12 weeks. The main outcome measures were body mass index, blood pressure, cardiorespiratory fitness, abdominal muscle strength, and low back and hamstring flexibility. Potential determinants of change in physical health were demographic variables, psychological variables (self-esteem, mastery, and kinesiophobia), and self-perceived health. RESULTS: The initial response was 73% and 252 persons had complete data collection at baseline. In total, 36 subjects were lost during follow-up. Participants were predominantly low educated, long-term unemployed, and in poor health. Participation in the programme was not influenced by demographic and psychological factors or by self-reported health. Drop-outs were younger and had a lower body mass index at baseline than subjects who completed the programme. At post-test, participants' cardiorespiratory fitness, abdominal muscle strength, and flexibility had increased by 6.8%-51.0%, whereas diastolic and systolic blood pressures had decreased by 2.2%-2.5%. The effect sizes ranges from 0.17-0.68. CONCLUSION: Participants with the poorest physical health benefited most from the programme and gender differences in improvement were observed. Physical health of unemployed persons with health complaints improved after participation in this health promotion programme, but not sufficiently, considering their poor physical health at baseline. PMID- 19545363 TI - Serum resistin levels in critically ill patients are associated with inflammation, organ dysfunction and metabolism and may predict survival of non septic patients. AB - INTRODUCTION: Blood glucose levels and insulin resistance in critically ill patients on admission to intensive care units (ICUs) have been identified as factors influencing mortality. The pathogenesis of insulin resistance (IR) in critically ill patients is complex and not fully understood. Resistin is a hormone mainly derived from macrophages in humans and from adipose tissue in rodents, which regulates glucose metabolism and insulin sensitivity. In non critically ill patients, resistin was found to be related to impaired glucose tolerance, insulin resistance, metabolic syndrome, obesity and type 2 diabetes. Therefore, resistin might represent a link between inflammation, acute phase response and insulin resistance in critically ill patients. We aimed to examine the correlation of serum resistin concentrations to parameters of inflammation, organ function, metabolism, disease severity and survival in critically ill patients. METHODS: On admission to the Medical ICU, 170 patients (122 with sepsis, 48 without sepsis) were studied prospectively and compared with 60 healthy non-diabetic controls. Clinical data, various laboratory parameters, metabolic and endocrine functions as well as investigational inflammatory cytokine profiles were assessed. Patients were followed for approximately three years. RESULTS: Resistin serum concentrations were significantly elevated in all critical care patients compared with healthy controls, and significantly higher in sepsis than in non-sepsis patients. Serum resistin concentrations were not associated with pre-existing type 2 diabetes or obesity. For all critically ill patients, a correlation to the homeostasis model assessment index of insulin resistance (HOMA-IR) was shown. Serum resistin concentrations were closely correlated to inflammatory parameters such as C-reactive protein, leukocytes, procalcitonin, and cytokines such as IL6 and TNF-alpha, as well as associated with renal failure and liver synthesis capacity. High resistin levels (> 10 ng/ml) were associated with an unfavourable outcome in non-sepsis patients on ICU and the overall survival. CONCLUSIONS: Serum resistin concentrations are elevated in acute inflammation due to sepsis or systemic inflammatory response syndrome (SIRS). The close correlation with other acute phase proteins suggests a predominant, clinically relevant resistin release from macrophages in ICU patients. Moreover, resistin could potentially serve as a prognostic biomarker in non-sepsis critically ill patients. PMID- 19545365 TI - Negative predictive value and potential cost savings of acute nuclear myocardial perfusion imaging in low risk patients with suspected acute coronary syndrome: a prospective single blinded study. AB - BACKGROUND: Previous studies from the USA have shown that acute nuclear myocardial perfusion imaging (MPI) in low risk emergency department (ED) patients with suspected acute coronary syndrome (ACS) can be of clinical value. The aim of this study was to evaluate the utility and hospital economics of acute MPI in Swedish ED patients with suspected ACS. METHODS: We included 40 patients (mean age 55 +/- 2 years, 50% women) who were admitted from the ED at Lund University Hospital for chest pain suspicious of ACS, and who had a normal or non-ischemic ECG and no previous myocardial infarction. All patients underwent MPI from the ED, and the results were analyzed only after patient discharge. The current diagnostic practice of admitting the included patients for observation and further evaluation was compared to a theoretical "MPI strategy", where patients with a normal MPI test would have been discharged home from the ED. RESULTS: Twenty-seven patients had normal MPI results, and none of them had ACS. MPI thus had a negative predictive value for ACS of 100%. With the MPI strategy, 2/3 of the patients would thus have been discharged from the ED, resulting in a reduction of total hospital cost by some 270 EUR and of bed occupancy by 0.8 days per investigated patient. CONCLUSION: Our findings in a Swedish ED support the results of larger American trials that acute MPI has the potential to safely reduce the number of admissions and decrease overall costs for low-risk ED patients with suspected ACS. PMID- 19545366 TI - Identifying strategies to maximise recruitment and retention of practices and patients in a multicentre randomised controlled trial of an intervention to optimise secondary prevention for coronary heart disease in primary care. AB - BACKGROUND: Recruitment and retention of patients and healthcare providers in randomised controlled trials (RCTs) is important in order to determine the effectiveness of interventions. However, failure to achieve recruitment targets is common and reasons why a particular recruitment strategy works for one study and not another remain unclear. We sought to describe a strategy used in a multicentre RCT in primary care, to report researchers' and participants' experiences of its implementation and to inform future strategies to maximise recruitment and retention. METHODS: In total 48 general practices and 903 patients were recruited from three different areas of Ireland to a RCT of an intervention designed to optimise secondary prevention of coronary heart disease. The recruitment process involved telephoning practices, posting information, visiting practices, identifying potential participants, posting invitations and obtaining consent. Retention involved patients attending reviews and responding to questionnaires and practices facilitating data collection. RESULTS: We achieved high retention rates for practices (100%) and for patients (85%) over an 18-month intervention period. Pilot work, knowledge of the setting, awareness of change in staff and organisation amongst participant sites, rapid responses to queries and acknowledgement of practitioners' contributions were identified as being important. Minor variations in protocol and research support helped to meet varied, complex and changing individual needs of practitioners and patients and encouraged retention in the trial. A collaborative relationship between researcher and practice staff which required time to develop was perceived as vital for both recruitment and retention. CONCLUSION: Recruiting and retaining the numbers of practices and patients estimated as required to provide findings with adequate power contributes to increased confidence in the validity and generalisability of RCT results. A continuous dynamic process of monitoring progress within trials and tailoring strategies to particular circumstances, whilst not compromising trial protocols, should allow maximal recruitment and retention. TRIAL REGISTRATION: ISRCTN24081411. PMID- 19545368 TI - Expression of p-AKT characterizes adenoid cystic carcinomas of head and neck with a higher risk for tumor relapses. AB - BACKGROUND: Adenoid cystic carcinomas are rare tumors with an indolent clinical course, but frequent local relapses. The identification of tumors with a higher relapse risk seems to be interesting. Hence we investigated parameters of glucose metabolism, which were found associated with poor prognosis in other malignancies. METHODS: Specimen of 29 patients were investigated immunohistochemically with antibodies against p-AKT, TKTL-1 (transketolase-like 1), M2PK (M2 pyruvate kinase), and GLUT-1. Proliferation was investigated by staining with Ki67. The tumors were located at the major or minor salivary glands. Only the typical cribriform subtype was investigated. The initial tumor stage was pT1 or pT2. RESULTS: Expression of p-AKT was significantly (P = 0.036) associated with a higher relapse risk in multivariate analysis. Low expression of M2PK was non-significantly (P = 0.065) predictive for a higher risk. TKTL-1 and GLUT-1 were expressed in the majority of cases, albeit not associated with relapse risk. CONCLUSION: Adenoid cystic carcinomas positive for p-AKT show a higher relapse risk. However, other parameters of glucose metabolism investigated here or proliferation (Ki67) were not predictive in this entity. Our findings demonstrate a possible background for therapeutic approaches targeting the inhibition of PI3K/AKT pathway. PMID- 19545367 TI - Transcriptional control of axonal guidance and sorting in dorsal interneurons by the Lim-HD proteins Lhx9 and Lhx1. AB - BACKGROUND: Lim-HD proteins control crucial aspects of neuronal differentiation, including subtype identity and axonal guidance. The Lim-HD proteins Lhx2/9 and Lhx1/5 are expressed in the dorsal spinal interneuron populations dI1 and dI2, respectively. While they are not required for cell fate acquisition, their role in patterning the axonal trajectory of dI1 and dI2 neurons remains incompletely understood. RESULTS: Using newly identified dI1- and dI2-specific enhancers to trace axonal trajectories originating from these interneurons, we found that each population is subdivided into several distinct groups according to their axonal pathways. dI1 neurons project axons rostrally, either ipsi- or contra-laterally, while dI2 are mostly commissural neurons that project their axons rostrally and caudally. The longitudinal axonal tracks of each neuronal population self fasciculate to form dI1- and dI2-specific bundles. The dI1 bundles are spatially located ventral relative to dI2 bundles. To examine the functional contribution of Lim-HD proteins to establishment of dI axonal projections, the Lim-HD code of dI neurons was altered by cell-specific ectopic expression. Expression of Lhx1 in dI1 neurons caused a repression of Lhx2/9 and imposed caudal projection to the caudal commissural dI1 neurons. Complementarily, when expressed in dI2 neurons, Lhx9 repressed Lhx1/5 and triggered a bias toward rostral projection in otherwise caudally projecting dI2 neurons, and ventral shift of the longitudinal axonal fascicule. CONCLUSION: The Lim-HD proteins Lhx9 and Lhx1 serve as a binary switch in controlling the rostral versus caudal longitudinal turning of the caudal commissural axons. Lhx1 determines caudal turning and Lhx9 triggers rostral turning. PMID- 19545369 TI - Incidence of breast cancer in Italy: mastectomies and quadrantectomies performed between 2000 and 2005. AB - OBJECTIVES: We aimed to determine the incidence of women's breast cancer in Italy without using statistical approximations. METHODS: We analyzed the national hospitalizations database at the Ministry of Health to calculate the number of major surgeries in Italian women (mastectomies and quadrantectomies) due to breast cancer between 2000 and 2005, overall and by age groups (<44, 45-64, 65-74 and >or= 75 years old). RESULTS: Over the six years examined, an overall number of 100,745 mastectomies and 168,147 quadrantectomies were performed. A total of 41,608 major surgeries due to breast cancer were performed in the year 2000 and this number rose to 47,200 in 2005, with a 13.4% increase over six years. CONCLUSION: by analyzing the hospitalizations database concerning major breast surgery, incidence of breast cancer in Italy was found to be 26.5% higher than the official estimations which have been computed using statistical models (namely 47,200 vs. 37,300 cases in year 2005). PMID- 19545370 TI - The HaDREB2 transcription factor enhances basal thermotolerance and longevity of seeds through functional interaction with HaHSFA9. AB - BACKGROUND: Transcription factor HaDREB2 was identified in sunflower (Helianthus annuus L.) as a drought-responsive element-binding factor 2 (DREB2) with unique properties. HaDREB2 and the sunflower Heat Shock Factor A9 (HaHSFA9) co-activated the Hahsp17.6G1 promoter in sunflower embryos. Both factors could be involved in transcriptional co-activation of additional small heat stress protein (sHSP) promoters, and thus contribute to the HaHSFA9-mediated enhancement of longevity and basal thermotolerance of seeds. RESULTS: We found that overexpression of HaDREB2 in seeds did not enhance longevity. This was deduced from assays of basal thermotolerance and controlled seed-deterioration, which were performed with transgenic tobacco. Furthermore, the constitutive overexpression of HaDREB2 did not increase thermotolerance in seedlings or result in the accumulation of HSPs at normal growth temperatures. In contrast, when HaDREB2 and HaHSFA9 were conjointly overexpressed in seeds, we observed positive effects on seed longevity, beyond those observed with overexpression of HaHSFA9 alone. Such additional effects are accompanied by a subtle enhancement of the accumulation of subsets of sHSPs belonging to the CI and CII cytosolic classes. CONCLUSION: Our results reveal the functional interdependency of HaDREB2 and HaHSFA9 in seeds. HaDREB2 differs from other previously characterized DREB2 factors in plants in terms of its unique functional interaction with the seed-specific HaHSFA9 factor. No functional interaction between HaDREB2 and HaHSFA9 was observed when both factors were conjointly overexpressed in vegetative tissues. We therefore suggest that additional, seed-specific factors, or protein modifications, could be required for the functional interaction between HaDREB2 and HaHSFA9. PMID- 19545371 TI - Generation and characterization of human insulin-releasing cell lines. AB - BACKGROUND: The in vitro culture of insulinomas provides an attractive tool to study cell proliferation and insulin synthesis and secretion. However, only a few human beta cell lines have been described, with long-term passage resulting in loss of insulin secretion. Therefore, we set out to establish and characterize human insulin-releasing cell lines. RESULTS: We generated ex-vivo primary cultures from two independent human insulinomas and from a human nesidioblastosis, all of which were cultured up to passage number 20. All cell lines secreted human insulin and C-peptide. These cell lines expressed neuroendocrine and islets markers, confirming the expression profile found in the biopsies. Although all beta cell lineages survived an anchorage independent culture, none of them were able to invade an extracellular matrix substrate. CONCLUSION: We have established three human insulin-releasing cell lines which maintain antigenic characteristics and insulin secretion profiles of the original tumors. These cell lines represent valuable tools for the study of molecular events underlying beta cell function and dysfunction. PMID- 19545373 TI - Mobile cell-phones (M-phones) in telemicroscopy: increasing connectivity of isolated laboratories. AB - BACKGROUND: The development of modern information telecommunication (ITC) technology and its use in telemedicine plays an increasingly important role in facilitating access to some diagnostic services even to people living in the most remote areas. However, physical and economical constraints in the access to broad band data-transmission network, still represent a considerable obstacle to the transmission of images for the purpose of tele-pathology. METHODS: Indifferently using m-phones of different brands, and a variety of microscopic preparations, images were taken without the use of any adaptor simply approaching the lens of the mobile cell phone camera to the ocular of common optical microscopes, and subsequently sent via Multimedia Messaging Services (MMS) to distant reference centres for tele-diagnosis. Access to MMS service was reviewed with specific reference to the African information communication technology (ICT) market. RESULTS: Images of any pathologic preparation could be captured and sent over the mobile phone with an MMS, without being limited by appropriate access to the internet for transmission (i.e. access to broad-band services). The quality of the image was not influenced by the brand or model of the mobile-phone used, but only by its digital resolution, with any resolution above 0.8 megapixel resulting in images sufficient for diagnosis.Access to MMS services is increasingly reaching remote disadvantaged areas. Current penetration of the service in Africa was mapped appearing already available in almost every country, with penetration index varying from 1.5% to 92.2%. CONCLUSION: The use of otherwise already widely available technologies, without any need for adaptors or otherwise additional technology, could significantly increase opportunities and quality diagnostics while lowering costs and considerably increasing connectivity between most isolated laboratories and distant reference center. PMID- 19545372 TI - Platform dependence of inference on gene-wise and gene-set involvement in human lung development. AB - BACKGROUND: With the recent development of microarray technologies, the comparability of gene expression data obtained from different platforms poses an important problem. We evaluated two widely used platforms, Affymetrix U133 Plus 2.0 and the Illumina HumanRef-8 v2 Expression Bead Chips, for comparability in a biological system in which changes may be subtle, namely fetal lung tissue as a function of gestational age. RESULTS: We performed the comparison via sequence based probe matching between the two platforms. "Significance grouping" was defined as a measure of comparability. Using both expression correlation and significance grouping as measures of comparability, we demonstrated that despite overall cross-platform differences at the single gene level, increased correlation between the two platforms was found in genes with higher expression level, higher probe overlap, and lower p-value. We also demonstrated that biological function as determined via KEGG pathways or GO categories is more consistent across platforms than single gene analysis. CONCLUSION: We conclude that while the comparability of the platforms at the single gene level may be increased by increasing sample size, they are highly comparable ontologically even for subtle differences in a relatively small sample size. Biologically relevant inference should therefore be reproducible across laboratories using different platforms. PMID- 19545374 TI - Parallelization and optimization of genetic analyses in isolation by distance web service. AB - BACKGROUND: The Isolation by Distance Web Service (IBDWS) is a user-friendly web interface for analyzing patterns of isolation by distance in population genetic data. IBDWS enables researchers to perform a variety of statistical tests such as Mantel tests and reduced major axis regression (RMA), and returns vector based graphs. The more than 60 citations since 2005 confirm the popularity and utility of this website. Despite its usefulness, the data sets with over 65 populations can take hours or days to complete due to the computational intensity of the statistical tests. This is especially troublesome for web-based software analysis, since users tend to expect real-time results on the order of seconds, or at most, minutes. Moreover, as genetic data continue to increase and diversify, so does the demand for more processing power. In order to increase the speed and efficiency of IBDWS, we first determined which aspects of the code were most time consuming and whether they might be amenable to improvements by parallelization or algorithmic optimization. RESULTS: Runtime tests uncovered two areas of IBDWS that consumed significant amounts of time: randomizations within the Mantel test and the RMA calculations. We found that these sections of code could be restructured and parallelized to improve efficiency. The code was first optimized by combining two similar randomization routines, implementing a Fisher Yates shuffling algorithm, and then parallelizing those routines. Tests of the parallelization and Fisher-Yates algorithmic improvements were performed on a variety of data sets ranging from 10 to 150 populations. All tested algorithms showed runtime reductions and a very close fit to the predicted speedups based on time-complexity calculations. In the case of 150 populations with 10,000 randomizations, data were analyzed 23 times faster. CONCLUSION: Since the implementation of the new algorithms in late 2007, datasets have continued to increase substantially in size and many exceed the largest population sizes we used in our test sets. The fact that the website has continued to work well in "real-world" tests, and receives a considerable number of new citations provides the strongest testimony to the effectiveness of our improvements. However, we soon expect the need to upgrade the number of nodes in our cluster significantly as dataset sizes continue to expand. The parallel implementation can be found at http://ibdws.sdsu.edu/. PMID- 19545376 TI - GP-support by means of AGnES-practice assistants and the use of telecare devices in a sparsely populated region in Northern Germany--proof of concept. AB - BACKGROUND: In many rural regions in Germany, the proportion of the elderly population increases rapidly. Simultaneously, about one-third of the presently active GPs will retire until 2010. Often it is difficult to find successors for vacant GP-practices. These regions require innovative concepts to avoid the imminent shortage in primary health care.The AGnES-concept comprises the delegation of GP-home visits to qualified AGnES-practice assistants (AGnES: GP supporting, community-based, e-health-assisted, systemic intervention). Main objectives were the assessment of the acceptance of the AGnES-concept by the participating GPs, patients, and AGnES-practice assistants, the kind of delegated tasks, and the feasibility of home telecare in a GP-practice. METHODS: In this paper, we report first results of the implementation of this concept in regular GP-practices, conducted November 2005--March 2007 on the Island of Rugen, Mecklenburg-Western Pomerania, Germany. This study was meant as a proof of concept.The GP delegated routine home-visits to qualified practice employees (here: registered nurses). Eligible patients were provided with telecare-devices to monitor disease-related physiological values.All delegated tasks, modules conducted and questionnaire responses were documented. The participating patients were asked for their acceptance based on standardized questionnaires. The GPs and AGnES-practice assistants were asked for their judgement about different project components, the quality of health care provision and the competences of the AGnES practice assistants. RESULTS: 550 home visits were conducted. 105 patients, two GPs and three AGnES-practice assistants (all registered nurses) participated in the project. 48 patients used telecare-devices to monitor health parameters. 87.4% of the patients accepted AGnES-care as comparable to common GP-care. In the course of the project, the GPs delegated an increasing number of both monitoring and interventional tasks to the AGnES-practice assistants. The GPs agreed that delegating tasks to a qualified practice assistant relieves them in their daily work. CONCLUSION: A part of the GPs home visits can be delegated to AGnES practice assistants to support GPs in regions with an imminent or already existing undersupply in primary care. The project triggered discussions among the institutions involved in the German healthcare system and supported a reconciliation of the respective competences of physicians and other medical professions. PMID- 19545375 TI - Identifying alemtuzumab as an anti-myeloid cell antiangiogenic therapy for the treatment of ovarian cancer. AB - BACKGROUND: Murine studies suggest that myeloid cells such as vascular leukocytes (VLC) and Tie2+ monocytes play a critical role in tumor angiogenesis and vasculogenesis. Myeloid cells are a primary cause of resistance to anti-VEGF therapy. The elimination of these cells from the tumor microenvironment significantly restricts tumor growth in both spontaneous and xenograft murine tumor models. Thus animal studies indicate that myeloid cells are potential therapeutic targets for solid tumor therapy. Abundant VLC and Tie2+ monocytes have been reported in human cancer. Unfortunately, the importance of VLC in human cancer growth remains untested as there are no confirmed therapeutics to target human VLC. METHODS: We used FACS to analyze VLC in ovarian and non-ovarian tumors, and characterize the relationship of VLC and Tie2-monocytes. We performed qRT-PCR and FACS on human VLC to assess the expression of the CD52 antigen, the target of the immunotherapeutic Alemtuzumab. We assessed Alemtuzumab's ability to induce complement-mediated VLC killing in vitro and in human tumor ascites. Finally we assessed the impact of anti-CD52 immuno-toxin therapy on murine ovarian tumor growth. RESULTS: Human VLC are present in ovarian and non-ovarian tumors. The majority of VLC appear to be Tie2+ monocytes. VLC and Tie2+ monocytes express high levels of CD52, the target of the immunotherapeutic Alemtuzumab. Alemtuzumab potently induces complement-mediated lysis of VLC in vitro and ex vivo in ovarian tumor ascites. Anti-CD52 immunotherapy targeting VLC restricts tumor angiogenesis and growth in murine ovarian cancer. CONCLUSION: These studies confirm VLC/myeloid cells as therapeutic targets in ovarian cancer. Our data provide critical pre-clinical evidence supporting the use of Alemtuzumab in clinical trials to test its efficacy as an anti-myeloid cell antiangiogenic therapeutic in ovarian cancer. The identification of an FDA approved anti-VLC agent with a history of clinical use will allow immediate proof-of-principle clinical trials in patients with ovarian cancer. PMID- 19545377 TI - The global pharmacy workforce: a systematic review of the literature. AB - The importance of health workforce provision has gained significance and is now considered one of the most pressing issues worldwide, across all health professions. Against this background, the objectives of the work presented here were to systematically explore and identify contemporary issues surrounding expansion of the global pharmacy workforce in order to assist the International Pharmaceutical Federation working group on the workforce. International peer and non-peer-reviewed literature published between January 1998 and February 2008 was analysed. Articles were collated by performing searches of appropriate databases and reference lists of relevant articles; in addition, key informants were contacted. Information that met specific quality standards and pertained to the pharmacy workforce was extracted to matrices and assigned an evidence grade. Sixty-nine papers were identified for inclusion (48 peer reviewed and 21 non-peer reviewed). Evaluation of evidence revealed the global pharmacy workforce to be composed of increasing numbers of females who were working fewer hours; this decreased their overall full-time equivalent contribution to the workforce, compared to male pharmacists. Distribution of pharmacists was uneven with respect to location (urban/rural, less-developed/more-developed countries) and work sector (private/public). Graduates showed a preference for completing pre registration training near where they studied as an undergraduate; this was of considerable importance to rural areas. Increases in the number of pharmacy student enrollments and pharmacy schools occurred alongside an expansion in the number and roles of pharmacy technicians. Increased international awareness and support existed for the certification, registration and regulation of pharmacy technicians and accreditation of training courses. The most common factors adding to the demand for pharmacists were increased feminization, clinical governance measures, complexity of medication therapy and increased prescriptions. To maintain and expand the future pharmacy workforce, increases in recruitment and retention will be essential, as will decreases in attrition, where possible. However, scaling up the global pharmacy workforce is a complex, multifactorial responsibility that requires coordinated action. Further research by means of prospective and comparative methods, not only surveys, is needed into feminization; decreasing demand for postgraduate training; graduate trends; job satisfaction and the impact of pharmacy technicians; and how effective existing interventions are at expanding the pharmacy workforce. More coordinated monitoring and modelling of the pharmacy workforce worldwide (particularly in developing countries) is required. PMID- 19545378 TI - Neurofibromatosis type 2 (NF2): a clinical and molecular review. AB - Neurofibromatosis type 2 (NF2) is a tumour-prone disorder characterised by the development of multiple schwannomas and meningiomas. Prevalence (initially estimated at 1: 200,000) is around 1 in 60,000. Affected individuals inevitably develop schwannomas, typically affecting both vestibular nerves and leading to hearing loss and deafness. The majority of patients present with hearing loss, which is usually unilateral at onset and may be accompanied or preceded by tinnitus. Vestibular schwannomas may also cause dizziness or imbalance as a first symptom. Nausea, vomiting or true vertigo are rare symptoms, except in late-stage disease. The other main tumours are schwannomas of the other cranial, spinal and peripheral nerves; meningiomas both intracranial (including optic nerve meningiomas) and intraspinal, and some low-grade central nervous system malignancies (ependymomas). Ophthalmic features are also prominent and include reduced visual acuity and cataract. About 70% of NF2 patients have skin tumours (intracutaneous plaque-like lesions or more deep-seated subcutaneous nodular tumours). Neurofibromatosis type 2 is a dominantly inherited tumour predisposition syndrome caused by mutations in the NF2 gene on chromosome 22. More than 50% of patients represent new mutations and as many as one-third are mosaic for the underlying disease-causing mutation. Although truncating mutations (nonsense and frameshifts) are the most frequent germline event and cause the most severe disease, single and multiple exon deletions are common. A strategy for detection of the latter is vital for a sensitive analysis. Diagnosis is based on clinical and neuroimaging studies. Presymptomatic genetic testing is an integral part of the management of NF2 families. Prenatal diagnosis and pre implantation genetic diagnosis is possible. The main differential diagnosis of NF2 is schwannomatosis. NF2 represents a difficult management problem with most patients facing substantial morbidity and reduced life expectancy. Surgery remains the focus of current management although watchful waiting with careful surveillance and occasionally radiation treatment have a role. Prognosis is adversely affected by early age at onset, a higher number of meningiomas and having a truncating mutation. In the future, the development of tailored drug therapies aimed at the genetic level are likely to provide huge improvements for this devastating condition. PMID- 19545380 TI - Pulmonary bullet embolism - a safe treatment strategy of a potentially fatal injury: a case report. AB - BACKGROUND: Vascular embolization of a projectile discharged from a weapon is a rare event. In this report a hunter's errant gunshot struck a farmer in the left chest. CASE REPORT: The projectile was lodged between the apex of the heart and the diaphragm. The patient was treated non-operatively and was discharged home only to return to the emergency department with chest pain and subsequent identification of the projectile in the left inferior pulmonary vein. Operative management consisted of a median sternotomy, cardiopulmonary bypass, and a pulmonary venectomy. CONCLUSION: He was subsequently discharged home and recovered uneventfully. PMID- 19545379 TI - Dealing with heterogeneity of treatment effects: is the literature up to the challenge? AB - BACKGROUND: Some patients will experience more or less benefit from treatment than the averages reported from clinical trials; such variation in therapeutic outcome is termed heterogeneity of treatment effects (HTE). Identifying HTE is necessary to individualize treatment. The degree to which heterogeneity is sought and analyzed correctly in the general medical literature is unknown. We undertook this literature sample to track the use of HTE analyses over time, examine the appropriateness of the statistical methods used, and explore the predictors of such analyses. METHODS: Articles were selected through a probability sample of randomized controlled trials (RCTs) published in Annals of Internal Medicine, BMJ, JAMA, The Lancet, and NEJM during odd numbered months of 1994, 1999, and 2004. RCTs were independently reviewed and coded by two abstractors, with adjudication by a third. Studies were classified as reporting: (1) HTE analysis, utilizing a formal test for heterogeneity or treatment-by-covariate interaction, (2) subgroup analysis only, involving no formal test for heterogeneity or interaction; or (3) neither. Chi-square tests and multiple logistic regression were used to identify variables associated with HTE reporting. RESULTS: 319 studies were included. Ninety-two (29%) reported HTE analysis; another 88 (28%) reported subgroup analysis only, without examining HTE formally. Major covariates examined included individual risk factors associated with prognosis, responsiveness to treatment, or vulnerability to adverse effects of treatment (56%); gender (30%); age (29%); study site or center (29%); and race/ethnicity (7%). Journal of publication and sample size were significant independent predictors of HTE analysis (p < 0.05 and p < 0.001, respectively). CONCLUSION: HTE is frequently ignored or incorrectly analyzed. An iterative process of exploratory analysis followed by confirmatory HTE analysis will generate the data needed to facilitate an individualized approach to evidence-based medicine. PMID- 19545381 TI - The DAWGPAWS pipeline for the annotation of genes and transposable elements in plant genomes. AB - BACKGROUND: High quality annotation of the genes and transposable elements in complex genomes requires a human-curated integration of multiple sources of computational evidence. These evidences include results from a diversity of ab initio prediction programs as well as homology-based searches. Most of these programs operate on a single contiguous sequence at a time, and the results are generated in a diverse array of readable formats that must be translated to a standardized file format. These translated results must then be concatenated into a single source, and then presented in an integrated form for human curation. RESULTS: We have designed, implemented, and assessed a Perl-based workflow named DAWGPAWS for the generation of computational results for human curation of the genes and transposable elements in plant genomes. The use of DAWGPAWS was found to accelerate annotation of 80-200 kb wheat DNA inserts in bacterial artificial chromosome (BAC) vectors by approximately twenty-fold and to also significantly improve the quality of the annotation in terms of completeness and accuracy. CONCLUSION: The DAWGPAWS genome annotation pipeline fills an important need in the annotation of plant genomes by generating computational evidences in a high throughput manner, translating these results to a common file format, and facilitating the human curation of these computational results. We have verified the value of DAWGPAWS by using this pipeline to annotate the genes and transposable elements in 220 BAC insertions from the hexaploid wheat genome (Triticum aestivum L.). DAWGPAWS can be applied to annotation efforts in other plant genomes with minor modifications of program-specific configuration files, and the modular design of the workflow facilitates integration into existing pipelines. PMID- 19545382 TI - Genotyping human ancient mtDNA control and coding region polymorphisms with a multiplexed Single-Base-Extension assay: the singular maternal history of the Tyrolean Iceman. AB - BACKGROUND: Progress in the field of human ancient DNA studies has been severely restricted due to the myriad sources of potential contamination, and because of the pronounced difficulty in identifying authentic results. Improving the robustness of human aDNA results is a necessary pre-requisite to vigorously testing hypotheses about human evolution in Europe, including possible admixture with Neanderthals. This study approaches the problem of distinguishing between authentic and contaminating sequences from common European mtDNA haplogroups by applying a multiplexed Single-Base-Extension assay, containing both control and coding region sites, to DNA extracted from the Tyrolean Iceman. RESULTS: The multiplex assay developed for this study was able to confirm that the Iceman's mtDNA belongs to a new European mtDNA clade with a very limited distribution amongst modern data sets. Controlled contamination experiments show that the correct results are returned by the multiplex assay even in the presence of substantial amounts of exogenous DNA. The overall level of discrimination achieved by targeting both control and coding region polymorphisms in a single reaction provides a methodology capable of dealing with most cases of homoplasy prevalent in European haplogroups. CONCLUSION: The new genotyping results for the Iceman confirm the extreme fallibility of human aDNA studies in general, even when authenticated by independent replication. The sensitivity and accuracy of the multiplex Single-Base-Extension methodology forms part of an emerging suite of alternative techniques for the accurate retrieval of ancient DNA sequences from both anatomically modern humans and Neanderthals. The contamination of laboratories remains a pressing concern in aDNA studies, both in the pre and post PCR environments, and the adoption of a forensic style assessment of a priori risks would significantly improve the credibility of results. PMID- 19545384 TI - The psychometric validation of a US English satisfaction measure for patients with benign prostatic hyperplasia and lower urinary tract symptoms. AB - BACKGROUND: The purpose of the current study was to validate the US English Patient Perception of Study Medication (PPSM) questionnaire, which measures patient satisfaction with Benign Prostatic Hyperplasia (BPH) treatment and was administered to men with BPH lower urinary tract symptoms (LUTS) enrolled in a multi-national clinical trial. METHODS: Patients with moderate to severe BPH symptoms completed three disease-specific measures: The International Prostate Symptom Score (IPSS), the BPH Impact Index (BII) and the PPSM, at baseline (after completion of the placebo run-in period) and at every 13-week clinic visit thereafter for the duration of the study treatment period. The PPSM was analysed to assess its variability, reliability and validity. RESULTS: There were 879 patients included in the analyses, with a mean age of 66.7 years. The PPSM was found to comprise two factors - PPSM-Global and PPSM-Pain, with a Total Score ranging from 7 to 49. It demonstrated good internal consistency (Cronbach's alpha ranged from .95 to .97) and also demonstrated convergent validity through significant correlations with the IPSS (.48 to .58), IPSS Quality of Life (QoL) item (.41 to .63) and BII (.31 to .45) and known-groups validity against the IPSS, IPSS QoL item and BII. CONCLUSION: Results support the use of the PPSM as a measure of satisfaction in BPH patient groups. PMID- 19545383 TI - Type-1 Collagen differentially alters beta-catenin accumulation in primary Dupuytren's Disease cord and adjacent palmar fascia cells. AB - BACKGROUND: Dupuytren's Disease (DD) is a debilitating contractile fibrosis of the palmar fascia characterised by excess collagen deposition, contractile myofibroblast development, increased transforming growth factor-beta levels and beta-catenin accumulation. The aim of this study was to determine if a collagen enriched environment, similar to in vivo conditions, altered beta-catenin accumulation by primary DD cells in the presence or absence of transforming growth factor-beta. METHODS: Primary DD and patient matched, phenotypically normal palmar fascia (PF) cells were cultured in the presence or absence of type 1 collagen and transforming growth factor-beta1. beta-catenin and alpha-smooth muscle actin levels were assessed by western immunoblotting and immunofluorescence microscopy. RESULTS: DD cells display a rapid depletion of cellular beta-catenin not evident in patient-matched PF cells. This effect was not evident in either cell type when cultured in the absence of type-1 collagen. Exogenous addition of transforming growth factor-beta1 to DD cells in collagen culture negates the loss of beta-catenin accumulation. Transforming growth factor beta1-induced alpha-smooth muscle actin, a marker of myofibroblast differentiation, is attenuated by the inclusion of type-1 collagen in cultures of DD and PF cells. CONCLUSION: Our findings implicate type-1 collagen as a previously unrecognized regulator of beta-catenin accumulation and a modifier of TGF-beta1 signaling specifically in primary DD cells. These data have implications for current treatment modalities as well as the design of in vitro models for research into the molecular mechanisms of DD. PMID- 19545385 TI - Structured chronic primary care and health-related quality of life in chronic heart failure. AB - BACKGROUND: Structured care is proposed as a lever for improving care for patients with chronic conditions. The purpose of this study was to explore the associations of structured care characteristics, derived from the Chronic Care Model, with health-related quality of life (HRQOL) and optimal clinical management in chronic heart failure (CHF) patients in primary care, as well as the association between optimal management and HRQOL. METHODS: Cross-sectional observational study using multi-level random-coefficient analyses of a representative sample of 357 patients diagnosed with CHF from 42 primary care practices in the Netherlands. We combined individual medical record data with patient and physician questionnaires. RESULTS: There was large variation in the levels and presence of structured care elements. A 91% of physicians indicated that next appointments for CHF patients were made immediately after visits, while 11% indicated that reminders on CHF management were periodically received in their practice. Few associations were found between the organizational characteristics and optimal treatment or HRQOL. Optimal pharmacological treatment related to better quality of life (beta = -11.5, P < .0001). Also, more lifestyle advice was given in practices with an appointment system allowing contact with more than one professional during the encounter (beta = 1.0, P = .04). CONCLUSION: HRQOL and treatment quality in CHF patients were not consistently associated with characteristics of structured care in primary care practices. PMID- 19545386 TI - Analyzing musculoskeletal neck pain, measured as present pain and periods of pain, with three different regression models: a cohort study. AB - BACKGROUND: In the literature there are discussions on the choice of outcome and the need for more longitudinal studies of musculoskeletal disorders. The general aim of this longitudinal study was to analyze musculoskeletal neck pain, in a group of young adults. Specific aims were to determine whether psychosocial factors, computer use, high work/study demands, and lifestyle are long-term or short-term factors for musculoskeletal neck pain, and whether these factors are important for developing or ongoing musculoskeletal neck pain. METHODS: Three regression models were used to analyze the different outcomes. Pain at present was analyzed with a marginal logistic model, for number of years with pain a Poisson regression model was used and for developing and ongoing pain a logistic model was used. Presented results are odds ratios and proportion ratios (logistic models) and rate ratios (Poisson model). The material consisted of web-based questionnaires answered by 1204 Swedish university students from a prospective cohort recruited in 2002. RESULTS: Perceived stress was a risk factor for pain at present (PR = 1.6), for developing pain (PR = 1.7) and for number of years with pain (RR = 1.3). High work/study demands was associated with pain at present (PR = 1.6); and with number of years with pain when the demands negatively affect home life (RR = 1.3). Computer use pattern (number of times/week with a computer session > or = 4 h, without break) was a risk factor for developing pain (PR = 1.7), but also associated with pain at present (PR = 1.4) and number of years with pain (RR = 1.2). Among life style factors smoking (PR = 1.8) was found to be associated to pain at present. The difference between men and women in prevalence of musculoskeletal pain was confirmed in this study. It was smallest for the outcome ongoing pain (PR = 1.4) compared to pain at present (PR = 2.4) and developing pain (PR = 2.5). CONCLUSION: By using different regression models different aspects of neck pain pattern could be addressed and the risk factors impact on pain pattern was identified. Short-term risk factors were perceived stress, high work/study demands and computer use pattern (break pattern). Those were also long-term risk factors. For developing pain perceived stress and computer use pattern were risk factors. PMID- 19545387 TI - Near-infrared spectroscopy-derived tissue oxygen saturation in battlefield injuries: a case series report. AB - BACKGROUND: Near-infrared spectroscopy technology has been utilized to monitor perfusion status in animal models of hemorrhagic shock and in human traumatic injury. To observe the effectiveness of such a device in a combat setting, an FDA approved device was used in conjunction with standard resuscitation and therapy of wounded patients presenting to the 228th Combat Support Hospital (CSH), Company B, over a three-month period. MATERIALS AND METHODS: These observations were performed on patients presenting to the 228th CSH, Co B, at Forward Operating Base Speicher, outside of Tikrit, Iraq, between the dates of June 15 and September 11, 2005. We utilized the Inspectra 325 tissue oxygen saturation (StO2) monitor (Hutchinson Technology, Inc; Hutchinson, MN, USA) with the probe placed on the thenar eminence or on another appropriate muscle bed, and used to monitor StO2 during early resuscitation and stabilization of patients. RESULTS: During the above time period, 161 patients were evaluated at the CSH as a result of traumatic injury and the device was placed on approximately 40 patients. In most patients, StO2 readings of greater than 70% were noted during the initial evaluation. No further information was collected from these patients. In 8 patients, convenience samples of StO2 data were collected along with pertinent physiologic data. In these patients, StO2 levels of below 70% tracked with hypotension, tachycardia, and clinical shock resulted in increases in StO2 after resuscitation maneuvers. CONCLUSION: Near-infrared spectroscopy-derived StO2 reflected and tracked the resuscitation status of our patients with battlefield injuries. StO2 has significant potential for use in resuscitation and care of patients with battlefield injuries. PMID- 19545388 TI - Effect of 6-months of physical exercise on the nitrate/nitrite levels in hypertensive postmenopausal women. AB - BACKGROUND: Evidences have showed that the incidence of arterial hypertension is greater in postmenopausal women as compared to premenopausal. Physical inactivity has been implicated as a major contributor to weight gain and abdominal obesity in postmenopausal women and the incidence of cardiovascular disease increases dramatically after menopause. Additionally, more women than men die each year of coronary heart disease and are twice as likely as men to die within the first year after a heart attack. A healthy lifestyle has been strongly associated with the regular physical activity and evidences have shown that physically active subjects have more longevity with reduction of morbidity and mortality. Nitric oxide (NO) produced by endothelial cells has been implicated in this beneficial effect with improvement of vascular relaxing and reduction in blood pressure in both laboratory animals and human. Although the effect of exercise training in the human cardiovascular system has been largely studied, the majority of these studies were predominantly conducted in men or young volunteers. Therefore, the aim of this work was to investigate the effects of 6 months of dynamic exercise training (ET) on blood pressure and plasma nitrate/nitrite concentration (NOx-) in hypertensive postmenopausal women. METHODS: Eleven volunteers were submitted to the ET consisting in 3 days a week, each session of 60 minutes during 6 months at moderate intensity (50% of heart rate reserve). Anthropometric parameters, blood pressure, NOx- concentration were measured at initial time and after ET. RESULTS: A significant reduction in both systolic and diastolic blood pressure values was seen after ET which was accompanied by markedly increase of NOx- levels (basal: 10 +/- 0.9; ET: 16 +/- 2 microM). Total cholesterol was significantly reduced (basal: 220 +/- 38 and ET: 178 +/- 22 mg/dl), whereas triglycerides levels were not modified after ET (basal: 141 +/- 89 and ET: 147 +/ 8 mg/dl). CONCLUSION: Our study shows that changing in lifestyle promotes reduction of arterial pressure which was accompanied by increase in nitrite/nitrate concentration. Therefore, 6-months of exercise training are an important approach in management arterial hypertension and play a protective effect in postmenopausal women. PMID- 19545389 TI - Prevalence of Khat chewing in college and secondary (high) school students of Jazan region, Saudi Arabia. AB - BACKGROUND: Khat is widely consumed among the youth of Jazan region of Saudi Arabia. However, its prevalence is not well documented. OBJECTIVE: This study was conducted to assess the prevalence and associated risk factors of khat chewing among college and secondary school students in Jazan region. METHODS: The study was conducted in May 2006 in the colleges and secondary schools in Jazan region. A sample of 10,000 students aged between 15 and 25 years was randomly selected. Students in each year of study were selected by systematic random sampling technique. Self-administered questionnaire was used for data collection. RESULTS: The overall prevalence of khat chewing in all the studied population was 21.4% (colleges 15.2% versus schools 21.5%). There were 3.8% female khat chewers and 37.70% male Khat chewers. Significant differences were found between khat chewers according to age, gender and residence (p < 0.05). The prevalence was different in different colleges and in different provinces of Jazan region. CONCLUSION: The prevalence of Khat chewing seems to be high among male students and not remarkable among female students. The use of Khat is significantly associated with age, gender, residence and school and college education (p < 0.05) among students of Jazan region. Strong measures need to be taken for greater awareness among school and college students to reduce its prevalence. PMID- 19545390 TI - Plasmodium falciparum genotypes diversity in symptomatic malaria of children living in an urban and a rural setting in Burkina Faso. AB - BACKGROUND: The clinical presentation of malaria, considered as the result of a complex interaction between parasite and human genetics, is described to be different between rural and urban areas. The analysis of the Plasmodium falciparum genetic diversity in children with uncomplicated malaria, living in these two different areas, may help to understand the effect of urbanization on the distribution of P. falciparum genotypes. METHODS: Isolates collected from 75 and 89 children with uncomplicated malaria infection living in a rural and an urban area of Burkina Faso, respectively, were analysed by a nested PCR amplification of msp1 and msp2 genes to compare P. falciparum diversity. RESULTS: The K1 allelic family was widespread in children living in the two sites, compared to other msp1 allelic families (frequency >90%). The MAD 20 allelic family of msp1 was more prevalent (p = 0.0001) in the urban (85.3%) than the rural area (63.2%). In the urban area, the 3D7 alleles of msp2 were more prevalent compared to FC27 alleles, with a high frequency for the 3D7 300bp allele (>30%). The multiplicity of infection was in the range of one to six in the urban area and of one to seven in the rural area. There was no difference in the frequency of multiple infections (p = 0.6): 96.0% (95% C.I: 91.6-100) in urban versus 93.1% (95%C.I: 87.6-98.6) in rural areas. The complexity of infection increased with age [p = 0.04 (rural area), p = 0.06 (urban area)]. CONCLUSION: Urban-rural area differences were observed in some allelic families (MAD20, FC27, 3D7), suggesting a probable impact of urbanization on genetic variability of P. falciparum. This should be taken into account in the implementation of malaria control measures. PMID- 19545391 TI - Adult B lymphoblastic leukaemia/lymphoma with hypodiploidy (-9) and a novel chromosomal translocation t(7;12)(q22;p13) presenting with severe eosinophilia - case report and review of literature. AB - Patients suffering from adult acute lymphoblastic leukemia are acutely ill and present most commonly with fever, pallor, bleeding, lymphadenopathy, hepatosplenomegaly and presence of lymphoblasts in the peripheral blood and bone marrow. We describe a rare presentation of acute lymphoblastic leukemia, in a young adult male who had vague and minimal symptoms with mild splenomegaly. There was severe eosinophilia along with absence of blasts in the peripheral blood, and 40% blasts with increase in eosinophils in the bone marrow. The blasts were positive for common precursor B cell markers on flow cytometry. The patient had a unique cytogenetic abnormality t(7;12)(q22;p13),-9, not previously described in acute lymphoblastic leukemia. He was categorized as poor risk due to failure to achieve complete remission after induction with UK ALL XII chemotherapy. PMID- 19545392 TI - Primary non-Hodgkin's lymphoma of the infratemporal fossa: a rare case report. AB - BACKGROUND: The head and neck are two of the most common sites of extranodal non Hodgkin's lymphoma (NHL). However, primary tumors of the infratemporal fossa are infrequent, and NHL in this region is extremely rare. CASE PRESENTATION: We present a case of a 41-year-old female that presented with swelling in the right preauricular region that had persisted for the past two years. The patient was diagnosed as having a small lymphocytic NHL. She initially underwent chemo radiation but reported relapse. The tumor was excised and again the patient underwent chemotherapy. The patient remained symptomatic and developed a second primary squamous cell carcinoma in the right retromolar trigone. DISCUSSION AND CONCLUSION: We discussed NHL with an emphasis on extranodal manifestations. Extranodal NHL that is limited to a single site can be managed by surgery and regular follow up. To the best of our knowledge, this is only the second case of primary NHL of the infratemporal fossa to be reported in the literature. PMID- 19545393 TI - Normal right- and left ventricular volumes and myocardial mass in children measured by steady state free precession cardiovascular magnetic resonance. AB - BACKGROUND: Quantification of ventricular volume by steady state free precession (SSFP) cardiovascular magnetic resonance is accurate and reproducible. Normal values exist for adults, but are lacking for children.We sought to establish normal values for left and right ventricular volumes, mass and function in healthy children by using SSFP. METHODS AND RESULTS: Fifty children (27 females, 23 males) without cardiovascular disease were evaluated. Median age was 11 years (range 7 months - 18 years), weight 35 kg (range 7-77 kg), height 146 cm (range 66-181 cm). Thirty-six examinations were performed with breath holding, 14 in freely breathing sedated children.Ventricular volumes and mass were measured in the end systolic and end diastolic phase on SSFP cine images acquired in a short axis plane as a stack of 12 contiguous slices covering full length of both ventricles. Regression analysis showed an exponential relationship between body surface area (BSA) and ventricular volumes and mass (normal value = a*BSAb). Normative curves for males and females are presented in relation to BSA for the end-diastolic volume, end-systolic volume and mass of both ventricles. Intra- and interobserver variability of the measurements was within the limits of 2% and 7% respectively, except for right ventricular mass (10%). CONCLUSION: The exponential equation for calculation of normal values for each ventricular parameter and graphical display of normative curves for data acquired in healthy children by SSFP cardiovascular magnetic resonance are provided. PMID- 19545394 TI - Anti-tumor effects of a human VEGFR-2-based DNA vaccine in mouse models. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) and its receptor, VEGFR-2 (Flk-1/KDR), play a key role in tumor angiogenesis. Blocking the VEGF-VEGFR-2 pathway may inhibit tumor growth. Here, we used human VEGFR-2 as a model antigen to explore the feasibility of immunotherapy with a plasmid DNA vaccine based on a xenogeneic homologue of this receptor. METHODS: The protective effects and therapeutic anti-tumor immunity mediated by the DNA vaccine were investigated in mouse models. Anti-angiogenesis effects were detected by immunohistochemical staining and the alginate-encapsulate tumor cell assay. The mechanism of action of the DNA vaccine was primarily explored by detection of auto-antibodies and CTL activity. RESULTS: The DNA vaccine elicited a strong, protective and therapeutic anti-tumor immunity through an anti-angiogenesis mechanism in mouse models, mediated by the stimulation of an antigen-specific response against mFlk-1. CONCLUSION: Our study shows that a DNA vaccine based on a xenogeneic homologue plasmid DNA induced autoimmunity against VEGFR-2, resulting in inhibition of tumor growth. Such vaccines may be clinically relevant for cancer immunotherapy. PMID- 19545395 TI - Role of complement and antibodies in controlling infection with pathogenic simian immunodeficiency virus (SIV) in macaques vaccinated with replication-deficient viral vectors. AB - BACKGROUND: We investigated the interplay between complement and antibodies upon priming with single-cycle replicating viral vectors (SCIV) encoding SIV antigens combined with Adeno5-SIV or SCIV pseudotyped with murine leukemia virus envelope boosting strategies. The vaccine was applied via spray-immunization to the tonsils of rhesus macaques and compared with systemic regimens. RESULTS: Independent of the application regimen or route, viral loads were significantly reduced after challenge with SIVmac239 (p < 0.03) compared to controls. Considerable amounts of neutralizing antibodies were induced in systemic immunized monkeys. Most of the sera harvested during peak viremia exhibited a trend with an inverse correlation between complement C3-deposition on viral particles and plasma viral load within the different vaccination groups. In contrast, the amount of the observed complement-mediated lysis did not correlate with the reduction of SIV titres. CONCLUSION: The heterologous prime-boost strategy with replication-deficient viral vectors administered exclusively via the tonsils did not induce any neutralizing antibodies before challenge. However, after challenge, comparable SIV-specific humoral immune responses were observed in all vaccinated animals. Immunization with single cycle immunodeficiency viruses mounts humoral immune responses comparable to live-attenuated immunodeficiency virus vaccines. PMID- 19545397 TI - Titanium dioxide nanoparticles induce oxidative stress and DNA-adduct formation but not DNA-breakage in human lung cells. AB - Titanium dioxide (TiO2), also known as titanium (IV) oxide or anatase, is the naturally occurring oxide of titanium. It is also one of the most commercially used form. To date, no parameter has been set for the average ambient air concentration of TiO2 nanoparticles (NP) by any regulatory agency. Previously conducted studies had established these nanoparticles to be mainly non-cyto- and genotoxic, although they had been found to generate free radicals both acellularly (specially through photocatalytic activity) and intracellularly. The present study determines the role of TiO2-NP (anatase, slashed circle < 100 nm) using several parameters such as cyto- and genotoxicity, DNA-adduct formation and generation of free radicals following its uptake by human lung cells in vitro. For comparison, iron containing nanoparticles (hematite, Fe2O3, slashed circle < 100 nm) were used. The results of this study showed that both types of NP were located in the cytosol near the nucleus. No particles were found inside the nucleus, in mitochondria or ribosomes. Human lung fibroblasts (IMR-90) were more sensitive regarding cyto- and genotoxic effects caused by the NP than human bronchial epithelial cells (BEAS-2B). In contrast to hematite NP, TiO2-NP did not induce DNA-breakage measured by the Comet-assay in both cell types. Generation of reactive oxygen species (ROS) was measured acellularly (without any photocatalytic activity) as well as intracellularly for both types of particles, however, the iron-containing NP needed special reducing conditions before pronounced radical generation. A high level of DNA adduct formation (8-OHdG) was observed in IMR-90 cells exposed to TiO2-NP, but not in cells exposed to hematite NP. Our study demonstrates different modes of action for TiO2- and Fe2O3-NP. Whereas TiO2-NP were able to generate elevated amounts of free radicals, which induced indirect genotoxicity mainly by DNA-adduct formation, Fe2O3-NP were clastogenic (induction of DNA-breakage) and required reducing conditions for radical formation. PMID- 19545396 TI - Perinatal transmission of human papilomavirus DNA. AB - The purpose was to study the perinatal transmission of human papillomavirus DNA (HPV-DNA) in 63 mother-newborn pairs, besides looking at the epidemiological factors involved in the viral DNA transmission. The following sampling methods were used: (1) in the pregnant woman, when was recruited, in cervix and clinical lesions of the vagina, vulva and perineal region; (2) in the newborn, (a) buccal, axillary and inguinal regions; (b) nasopharyngeal aspirate, and (c) cord blood; (3) in the children, buccal was repeated in the 4th week and 6th and 12th month of life. HPV-DNA was identified using two methodologies: multiplex PCR (PGMY09 and MY11 primers) and nested-PCR (genotypes 6/11, 16, 18, 31, 33, 42, 52 and 58). Perinatal transmission was considered when concordance was found in type-specific HPV between mother/newborn or mother/child. HPV-DNA genital was detected in 49 pregnant women submitted to delivery. Eleven newborns (22.4%, n = 11/49) were HPV DNA positive. In 8 cases (16.3%, n = 8/49) there was type specific HPV concordance between mother/newborn samples. At the end of the first month of life three children (6.1%, n = 3/49) became HPV-DNA positive, while two remained positive from birth. In 3 cases (100%, n = 3/3) there was type specific HPV concordance between mother/newborn samples. In the 6th month, a child (2%, n = 1/49) had become HPV-DNA positive between the 1st and 6th month of life, and there was type specific HPV concordance of mother/newborn samples. All the HPV DNA positive children (22.4%, n = 11/49) at birth and at the end first month of life (6.1%, n = 3/49) became HPV-DNA negative at the age of 6 months. The HPV-DNA positive child (2%, n = 1/49) from 1st to the 6th month of life became HPV-DNA negative between the 6th and 12th month of life and one child had anogenital warts. In the twelfth month all (100%, n = 49/49) the children studied were HPV DNA negative. A positive and significant correlation was observed between perinatal transmission of HPV-DNA and the immunodepression of maternal variables (HIV, p = 0.007). Finally, the study suggests that perinatal transmission of HPV DNA occurred in 24.5% (n = 12/49) of the cases studied. PMID- 19545398 TI - Task shifting: the answer to the human resources crisis in Africa? AB - Ever since the 2006 World Health Report advocated increased community participation and the systematic delegation of tasks to less-specialized cadres, there has been a great deal of debate about the expediency, efficacy and modalities of task shifting. The delegation of tasks from one cadre to another, previously often called substitution, is not a new concept. It has been used in many countries and for many decades, either as a response to emergency needs or as a method to provide adequate care at primary and secondary levels, especially in understaffed rural facilities, to enhance quality and reduce costs. However, rapidly increasing care needs generated by the HIV/AIDS epidemic and accelerating human resource crises in many African countries have given the concept and practice of task shifting new prominence and urgency. Furthermore, the question arises as to whether task shifting and increased community participation can be more than a short-term solution to address the HIV/AIDS crisis and can contribute to a revival of the primary health care approach as an answer to health systems crises. In this commentary we argue that, while task shifting holds great promise, any long-term success of task shifting hinges on serious political and financial commitments. We reason that it requires a comprehensive and integrated reconfiguration of health teams, changed scopes of practice and regulatory frameworks and enhanced training infrastructure, as well as availability of reliable medium- to long-term funding, with time frames of 20 to 30 years instead of three to five years. The concept and practice of community participation needs to be revisited. Most importantly, task shifting strategies require leadership from national governments to ensure an enabling regulatory framework; drive the implementation of relevant policies; guide and support training institutions and ensure adequate resources; and harness the support of the multiple stakeholders. With such leadership and a willingness to learn from those with relevant experience (for example, Brazil, Ethiopia, Malawi, Mozambique and Zambia), task shifting can indeed make a vital contribution to building sustainable, cost effective and equitable health care systems. Without it, task shifting runs the risk of being yet another unsuccessful health sector reform initiative. PMID- 19545400 TI - Salivary cortisol differs with age and sex and shows inverse associations with WHR in Swedish women: a cross-sectional study. AB - BACKGROUND: Most studies on cortisol have focused on smaller, selected samples. We therefore aimed to sex-specifically study the diurnal cortisol pattern and explore its association with abdominal obesity in a large unselected population. METHODS: In 2001-2004, 1811 men and women (30-75 years) were randomly selected from the Vara population, south-western Sweden (81% participation rate). Of these, 1671 subjects with full information on basal morning and evening salivary cortisol and anthropometric measurements were included in this cross-sectional study. Differences between groups were examined by general linear model and by logistic and linear regression analyses. RESULTS: Morning and Delta-cortisol (morning - evening cortisol) were significantly higher in women than men. In both genders older age was significantly associated with higher levels of all cortisol measures, however, most consistently with evening cortisol. In women only, age adjusted means of WHR were significantly lower in the highest compared to the lowest quartile of morning cortisol (p = 0.036) and Delta-cortisol (p < 0.001), respectively. Furthermore, when comparing WHR above and below the mean, the age adjusted OR in women for the lowest quartile of cortisol compared to the highest was 1.5 (1.0-2.2, p = 0.058) for morning cortisol and 1.9 (1.3-2.8) for Delta cortisol. All findings for Delta-cortisol remained after adjustments for multiple covariates and were also seen in a linear regression analysis (p = 0.003). CONCLUSION: In summary, our findings of generally higher cortisol levels in women than men of all ages are novel and the stronger results seen for Delta-cortisol as opposed to morning cortisol in the association with WHR emphasise the need of studying cortisol variation intra-individually. To our knowledge, the associations in this study have never before been investigated in such a large population sample of both men and women. Our results therefore offer important knowledge on the descriptive characteristics of cortisol in relation to age and gender, and on the impact that associations previously seen between cortisol and abdominal obesity in smaller, selected samples have on a population level. PMID- 19545399 TI - Development and preliminary evaluation of a quality of life measure targeted at dementia caregivers. AB - BACKGROUND: Providing care for individuals with a progressive, debilitating condition such as dementia can adversely impact the quality of life (QOL) of informal caregivers. To date, there is no existing caregiver quality of life measure for dementia caregivers with breadth of coverage or that is applicable to caregivers of diverse ethnic backgrounds. The purpose of this study was to develop and evaluate a caregiver-targeted quality-of-life measure (CGQOL) for informal caregivers of persons with dementia that can be used with caregivers from a variety of ethnicities. METHODS: 91 items were field tested by telephone interviews with 179 English-speaking and 21 monolingual Spanish-speaking caregivers of persons with dementia. Repeat interviews were conducted with 71 caregivers. Administration time, scale score distributions, item-scale correlations, reliability, and associations of scales with patient and caregiver demographic and caregiving characteristics were estimated. Structure of associations among scales was examined using exploratory factor analysis. RESULTS: Item analysis yielded 80 items distributed across 10 scales, with median administration time of 17 minutes [IQR 13.5-22 minutes] and minimal missing data. There were few floor or ceiling effects in scale score distributions. Internal consistency reliability was >or= 0.78 for all scales; test-retest reliability (intraclass correlation) estimates exceeded 0.70 for 6 scales. More hours weekly spent in caregiving was uniquely associated with worse quality of life on 8 scales (p's 0.05 for all). No detectable difference in IgM autoantibodies to the above antigens was observed between the two groups. CONCLUSIONS: GMT occurs in approximately 20.2% of LN patients. Patients with GMT have severer renal tissue injuries and poorer renal functions than patients without GMT. The lupus anticoagulant and antibodies against beta2 glycoprotein I and thrombin may play a role in GMT. PMID- 19545417 TI - The utility of MRI in predicting radiographic erosions in the metatarsophalangeal joints of the rheumatoid foot: a prospective longitudinal cohort study. AB - INTRODUCTION: Magnetic resonance imaging (MRI) may reveal rheumatoid arthritis (RA) changes in the feet when hands are normal. The purpose of this study was to determine the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of a metatarsophalangeal (MTP) erosion on MRI to predict a subsequent radiographic erosion in the same joint. Similar analyses were performed for bone marrow edema, predicting a subsequent MRI erosion. Descriptive results of other lesions are reported. METHODS: Fifty patients with RA of less than 5 years' duration who were rheumatoid factor-positive and/or anti cyclic citrullinated peptide-positive were recruited. Patients on anti-tumor necrosis factor (TNF) therapy were excluded. Anti-TNF therapy could begin after enrollment. MRI and radiographs of the 3rd, 4th, and 5th MTP joints bilaterally were taken at baseline and at 6, 12, and 24 months. Clinical data were collected. RESULTS: Fifty patients were recruited; 46 patients had suitable data. Results for MRI erosions predicting subsequent radiographic erosions for 6, 12, and 24 months, respectively, were as follows: sensitivity 0.75, 0.60, 0.75; specificity 0.93, 0.94, 0.94; PPV 0.086, 0.10, 0.17; NPV 0.998, 0.995, 0.995. Results for MRI bone marrow edema predicting MRI erosions at 6 and 12 months, respectively, revealed sensitivity 0.50, 0.67; specificity 0.97, 0.97; PPV 0.25, 0.50; NPV 0.99, 0.99. Synovitis was the most common finding and, when present in isolation, resolved on 67.3% of subsequent studies. MRI erosions persisted on subsequent studies with one exception. Forty-six percent of the cohort was on anti-TNF therapy after study inception. CONCLUSIONS: The PPV of MRI erosions to predict subsequent radiographic erosions was low. Similarly, the PPV of bone marrow edema to predict a later MRI erosion was low. Alternatively, the NPV of the absence of an MRI erosion or bone marrow edema predicts that a later radiographic erosion or MRI erosion will likely not develop. Anti-TNF therapies may have resulted in the lower-than-anticipated PPVs. MRI descriptions of bone edema may represent a more critical time to treat in order to avoid damage, whereas an MRI erosion represents more permanent damage. This study suggests that imaging modalities more sensitive than radiographs are necessary to monitor disease in the biologic era. PMID- 19545418 TI - A strategy for enrichment of claudins based on their affinity to Clostridium perfringens enterotoxin. AB - BACKGROUND: Claudins, a family of protein localized in tight junctions, are essential for the control of paracellular permeation in epithelia and endothelia. The interaction of several claudins with Clostridium perfringens enterotoxin (CPE) has been exploited for an affinity-based enrichment of CPE-binding claudins from lysates of normal rat cholangiocytes. RESULTS: Immunoblotting and mass spectrometry (MS) experiments demonstrate strong enrichment of the CPE-binding claudins -3, -4 and -7, indicating specific association with glutathione-S transferase (GST)-CPE(116-319) fusion protein. In parallel, the co-elution of (non-CPE-binding) claudin-1 and claudin-5 was observed. The complete set of co enriched proteins was identified by MS after electrophoretic separation. Relative mass spectrometric protein quantification with stable isotope labeling with amino acids in cell culture (SILAC) made it possible to discriminate specific binding from non-specific association to GST and/or matrix material. CONCLUSION: CPE(116 319) provides an efficient tool for single step enrichment of different claudins from cell lysates. Numerous proteins were shown to be co-enriched with the CPE binding claudins, but there are no indications (except for claudins -1 and -5) for an association with tight junctions. PMID- 19545419 TI - Influence of conjugated linoleic acid on the porcine immune response and morbidity: a randomized controlled trial. AB - BACKGROUND: Conjugated linoleic acid (CLA) has diverse influences on the immune response in different experimental models. In the present study we investigated the effect of CLA feeding on inflammatory and immune responses in a piglet model. We studied the duration of this effect and possible detrimental effects of CLA feeding. After 12 weeks of CLA and control supplementation and washout, animals were sacrificed and parenchymal organs were histologically examined. RESULTS: In activated peripheral mononuclear cells interferon-gamma was significantly (p = 0.008) lower in the CLA group by the end of the feeding period. This effect disappeared as soon as supplementation was stopped. No differences were found in the tumour necrosis factor-alpha, interleukin-10 production, serum immunoglobulin G levels and fat infiltration of the liver, except that fat storage cell infiltration was significantly (p < 0.04) higher in the CLA-fed group. The effect of time for interferon-gamma, interleukin-10 and immunoglobulin-G levels was statistically significant. CONCLUSION: At the end of the feeding period the interferon-gamma response was depressed. However, the maturation of the piglet immune system in our young pig model probably outweighs the impact of CLA feeding on the immune response, even though liver fat storage cell infiltration, which plays an important role in liver regeneration, increased during CLA feeding of the piglets. PMID- 19545420 TI - Improved resolution in the position of drought-related QTLs in a single mapping population of rice by meta-analysis. AB - BACKGROUND: Meta-analysis of QTLs combines the results of several QTL detection studies and provides narrow confidence intervals for meta-QTLs, permitting easier positional candidate gene identification. It is usually applied to multiple mapping populations, but can be applied to one. Here, a meta-analysis of drought related QTLs in the Bala x Azucena mapping population compiles data from 13 experiments and 25 independent screens providing 1,650 individual QTLs separated into 5 trait categories; drought avoidance, plant height, plant biomass, leaf morphology and root traits. A heat map of the overlapping 1 LOD confidence intervals provides an overview of the distribution of QTLs. The programme BioMercator is then used to conduct a formal meta-analysis at example QTL clusters to illustrate the value of meta-analysis of QTLs in this population. RESULTS: The heat map graphically illustrates the genetic complexity of drought related traits in rice. QTLs can be linked to their physical position on the rice genome using Additional file 1 provided. Formal meta-analysis on chromosome 1, where clusters of QTLs for all trait categories appear close, established that the sd1 semi-dwarfing gene coincided with a plant height meta-QTL, that the drought avoidance meta-QTL was not likely to be associated with this gene, and that this meta-QTL was not pleiotropic with close meta-QTLs for leaf morphology and root traits. On chromosome 5, evidence suggests that a drought avoidance meta QTL was pleiotropic with leaf morphology and plant biomass meta-QTLs, but not with meta-QTLs for root traits and plant height 10 cM lower down. A region of dense root QTL activity graphically visible on chromosome 9 was dissected into three meta-QTLs within a space of 35 cM. The confidence intervals for meta-QTLs obtained ranged from 5.1 to 14.5 cM with an average of 9.4 cM, which is approximately 180 genes in rice. CONCLUSION: The meta-analysis is valuable in providing improved ability to dissect the complex genetic structure of traits, and distinguish between pleiotropy and close linkage. It also provides relatively small target regions for the identification of positional candidate genes. PMID- 19545421 TI - KIF11 inhibition for glioblastoma treatment: reason to hope or a struggle with the brain? AB - BACKGROUND: Glioblastomas (GBM) are typically comprised of morphologically diverse cells. Despite current advances in therapy, including surgical resection followed by radiation and chemotherapy, the prognosis for patients with GBM remains poor. Unfortunately, most patients die within 2 years of diagnosis of their disease. Molecular abnormalities vary among individual patients and also within each tumor. Indeed, one of the distinguishing features of GBM is its marked genetic heterogeneity. Due to the brain location of the tumor, the potential target inhibition for anticancer therapy must exhibit a manageable neurotoxicity profile in the concentration range in which the compounds show anti proliferative activity.Kinesin KIF11 inhibition by small molecules such as Monastrol or Ispinesib is currently under investigation in the field of malignant tumors. In the current study we have assessed the relevance of the anti-mitotic Kinesin-like protein KIF11 in human GBM cell-lines. RESULTS: In this study the target was validated using a set of well characterised and potentially specific small molecule inhibitors of KIF11: an ispinesib analog, Monastrol, a Merck compound and 3 simplified derivatives of the Merck compound. Following an in silico selection, those compounds predicted to bear a favorable BBB permeation profile were assessed for their phenotypic effect on cell lines derived both from primary (U87MG) as well as treated (DBTRG-05-MG) glioblastomas. For some compounds, these data could be compared to their effect on normal human astrocytes, as well as their neurotoxicity on primary rat cortical neurons. The ispinesib analogue 1 showed an anti-proliferative effect on GBM cell lines by blocking them in the G2/M phase in a concentration range which was shown to be harmless to primary rat cortical neurons. Furthermore, ispinesib analog increased caspase 3/7-induced apoptosis in U87MG cells. CONCLUSION: In the area of cell cycle inhibition, KIF11 is critical for proper spindle assembly and represents an attractive anticancer target. Our results suggest that KIF11 inhibitors, when able to permeate the blood-brain-barrier, could represent an interesting class of anticancer drugs with low neurotoxic effects in the treatment of brain tumors. PMID- 19545422 TI - Primary stroke prevention for sickle cell disease in north-east Italy: the role of ethnic issues in establishing a Transcranial Doppler screening program. AB - BACKGROUND: Stroke is a serious complication of sickle cell disease (SCD) in children. Transcranic Doppler (TCD) is a well-established predictor of future cerebrovascular symptoms: a blood flow velocity >200 cm/sec in the Middle Cerebral Artery (MCA) correlates with a high risk of stroke in cohorts of African american HbS/HbS patients. In North-East Italy the recent increase in SCD patients is mainly due to immigration from Africa. A comprehensive care program for children with SCD was established in our Center since 2004, but a wide and routine screening for Primary stroke prevention needs to be developed. METHODS: In order to verify the feasibility of TCD and Transcranial color coded Sonography (TCCS) screening in our setting and the applicability of international reference values of blood velocities to our population of African immigrants with HbS/HbS SCD, we performed TCD and TCCD in 12 HbS/HbS African children and two groups of age-matched controls of Caucasian and African origin respectively. TCD and TCCS were performed on the same day of the scheduled routine hematologic visit after parental education. RESULTS: All parents accepted to perform the sonography to their children. TCD and TCCD were performed in all patients and an adequate temporal window could be obtained in all of them. Pulsatility index and depth values in both the MCA and the Basilar Artery (BA) were similar at TCD and TCCS evaluation in the three groups while time-average maximum velocities (TAMM), peak systolic velocity and diastolic velocity in the MCA and BA were higher in the patients' group on both TCD and TCCS evaluation. African and Caucasian healthy controls had similar lower values. CONCLUSION: Our preliminary data set the base to further evaluate the implementation of a primary stroke prevention program in our setting of HbS/HbS African immigrants and HbS/beta thalassemia Italians. Parental education-preferably in the native language- on stroke risk and prevention in SCD increases compliance and should be a necessary part of the program. Ethnic background does not seem to influence TCD velocity and internationally accepted reference values already validated in African-American SCD pediatric patients can be used, but long prospective trials are needed to verify their efficacy in defining stroke risk in our setting. PMID- 19545423 TI - Gold nanoparticles induce cytotoxicity in the alveolar type-II cell lines A549 and NCIH441. AB - BACKGROUND: During the last years engineered nanoparticles (NPs) have been extensively used in different technologies and consequently many questions have arisen about the risk and the impact on human health following exposure to nanoparticles. Nevertheless, at present knowledge about the cytotoxicity induced by NPs is still largely incomplete. In this context, we have investigated the cytotoxicity induced by gold nanoparticles (AuNPs), which differed in size and purification grade (presence or absence of sodium citrate residues on the particle surface) in vitro, in the human alveolar type-II (ATII)-like cell lines A549 and NCIH441. RESULTS: We found that the presence of sodium citrate residues on AuNPs impaired the viability of the ATII-like cell lines A549 and NCIH441. Interestingly, the presence of an excess of sodium citrate on the surface of NPs not only reduced the in vitro viability of the cell lines A549 and NCIH441, as shown by MTT assay, but also affected cellular proliferation and increased the release of lactate dehydrogenase (LDH), as demonstrated by Ki-67 and LDH-release assays respectively. Furthermore, we investigated the internalization of AuNPs by transmission electron microscopy (TEM) and we observed that particles were internalized by active endocytosis in the cell lines A549 and NCIH441 within 3 hr. In addition, gold particles accumulated in membrane-bound vesicles and were not found freely dispersed in the cytoplasm. CONCLUSION: Our data suggest that the presence of contaminants, such as sodium citrate, on the surface of gold nanoparticles might play a pivotal role in inducing cytotoxicity in vitro, but does not influence the uptake of the particles in human ATII-like cell lines. PMID- 19545424 TI - Protein Never in Mitosis Gene A Interacting-1 regulates calpain activity and the degradation of cyclooxygenase-2 in endothelial cells. AB - BACKGROUND: The peptidyl-proline isomerase, Protein Never in Mitosis Gene A Interacting-1 (PIN1), regulates turnover of inducible nitric oxide synthase (iNOS) in murine aortic endothelial cells (MAEC) stimulated with E. coli endotoxin (LPS) and interferon-gamma (IFN). Degradation of iNOS was reduced by a calpain inhibitor, suggesting that PIN1 may affect induction of other calpain sensitive inflammatory proteins, such as cyclooxygenase (COX)-2, in MAEC. METHODS: MAEC, transduced with lentivirus encoding an inactive control short hairpin (sh) RNA or one targeting PIN1 that reduced PIN1 by 85%, were used. Cells were treated with LPS/IFN, calpain inhibitors (carbobenzoxy-valinyl phenylalaninal (zVF), PD150606), cycloheximide and COX inhibitors to determine the effect of PIN1 depletion on COX-2 and calpain. RESULTS: LPS or IFN alone did not induce COX-2. However, treatment with 10 mug LPS plus 20 ng IFN per ml induced COX-2 protein 10-fold in Control shRNA MAEC. Induction was significantly greater (47-fold) in PIN1 shRNA cells. COX-2-dependent prostaglandin E2 production increased 3-fold in KD MAEC, but did not increase in Control cells. The additional increase in COX-2 protein due to PIN1 depletion was post transcriptional, as induction of COX-2 mRNA by LPS/IFN was the same in cells containing or lacking PIN1. Instead, the loss of COX-2 protein, after treatment with cycloheximide to block protein synthesis, was reduced in cells lacking PIN1 in comparison with Control cells, indicating that degradation of the enzyme was reduced. zVF and PD150606 each enhanced the induction of COX-2 by LPS/IFN. zVF also slowed the loss of COX-2 after treatment with cycloheximide, and COX-2 was degraded by exogenous mu-calpain in vitro. In contrast to iNOS, physical interaction between COX-2 and PIN1 was not detected, suggesting that effects of PIN1 on calpain, rather than COX-2 itself, affect COX-2 degradation. While cathepsin activity was unaltered, depletion of PIN1 reduced calpain activity by 55% in comparison with Control shRNA cells. CONCLUSION: PIN1 reduced calpain activity and slowed the degradation of COX-2 in MAEC, an effect recapitulated by an inhibitor of calpain. Given the sensitivity of COX-2 and iNOS to calpain, PIN1 may normally limit induction of these and other calpain substrates by maintaining calpain activity in endothelial cells. PMID- 19545425 TI - Altered effector function of peripheral cytotoxic cells in COPD. AB - BACKGROUND: There is mounting evidence that perforin and granzymes are important mediators in the lung destruction seen in COPD. We investigated the characteristics of the three main perforin and granzyme containing peripheral cells, namely CD8+ T lymphocytes, natural killer (NK; CD56+CD3-) cells and NKT like (CD56+CD3+) cells. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated and cell numbers and intracellular granzyme B and perforin were analysed by flow cytometry. Immunomagnetically selected CD8+ T lymphocytes, NK (CD56+CD3-) and NKT-like (CD56+CD3+) cells were used in an LDH release assay to determine cytotoxicity and cytotoxic mechanisms were investigated by blocking perforin and granzyme B with relevant antibodies. RESULTS: The proportion of peripheral blood NKT-like (CD56+CD3+) cells in smokers with COPD (COPD subjects) was significantly lower (0.6%) than in healthy smokers (smokers) (2.8%, p < 0.001) and non-smoking healthy participants (HNS) (3.3%, p < 0.001). NK (CD56+CD3-) cells from COPD subjects were significantly less cytotoxic than in smokers (16.8% vs 51.9% specific lysis, p < 0.001) as were NKT-like (CD56+CD3+) cells (16.7% vs 52.4% specific lysis, p < 0.001). Both cell types had lower proportions expressing both perforin and granzyme B. Blocking the action of perforin and granzyme B reduced the cytotoxic activity of NK (CD56+CD3-) and NKT-like (CD56+CD3+) cells from smokers and HNS. CONCLUSION: In this study, we show that the relative numbers of peripheral blood NK (CD56+CD3-) and NKT-like (CD56+CD3+) cells in COPD subjects are reduced and that their cytotoxic effector function is defective. PMID- 19545426 TI - Crotaline Fab antivenom appears to be effective in cases of severe North American pit viper envenomation: an integrative review. AB - BACKGROUND: In 2000, the United States Food and Drug Administration approved Crotalidae Polyvalent Immune Fab (Ovine) (hereafter, FabAV), "for the management of patients with minimal to moderate North American Crotalid envenomation." Because whole-IgG pit viper antivenom is no longer available in the United States, FabAV is currently the only specific treatment option available to United States clinicians treating snakebite victims of any severity. No clinical trial data are available concerning the effectiveness of FabAV for treatment of severe snakebite, but several published articles describe its use in this setting. METHODS: We performed a comprehensive review of the English-language medical literature to identify all publications (1996 to July, 2008) containing data about the administration of FabAV. Two trained reviewers separately extracted case-level data concerning the administration of FabAV to patients with severe envenomation by North American crotaline snakes to a standardized form. Descriptive statistics were used. In addition, we hand-searched the US National Poison Data System reports for the years 2000-2006 to identify and describe any reports of death that occurred after FabAV administration. RESULTS: The literature review found 147 unique publications regarding FabAV. Twenty-four evaluable cases of severe human envenomation treated with FabAV were identified in 19 publications. Seven cases were described in five cohort studies, and 17 cases were described in 14 single patient case reports or non-cohort case series. Sixty-five specific severe venom effects were reported in these 24 patients, of which 50 effects (77%) improved or resolved after FabAV therapy. Initial control of all severe venom effects was achieved in 12 patients (50%). The rate at which initial control was achieved was significantly higher among patients reported in the cohort series than in the case series and non-cohort reports (100% vs. 29%, P = 0.005). The median dose of FabAV used to obtain initial control was 6 vials (range: 4 - 18 vials). Nine patients had severe venom effects that persisted despite FabAV therapy. Recurrent and/or delayed-onset severe defibrination syndrome occurred in 12 patients, most of whom did not receive recommended maintenance FabAV dosing. No patient developed systemic bleeding. CONCLUSION: In this structured literature review, FabAV appears to be effective in the management of severe crotaline snake envenomation. Incomplete response to therapy, recurrence of venom effects, and delayed-onset venom effects were reported in case reports, but not reported in cohort studies. PMID- 19545427 TI - Does a monetary incentive improve the response to a postal questionnaire in a randomised controlled trial? The MINT incentive study. AB - BACKGROUND: Sending a monetary incentive with postal questionnaires has been found to improve the proportion of responders, in research in non-healthcare settings. However, there is little research on use of incentives to improve follow-up rates in clinical trials, and existing studies are inconclusive. We conducted a randomised trial among participants in the Managing Injuries of the Neck Trial (MINT) to investigate the effects on the proportion of questionnaires returned and overall non-response of sending a 5 pounds gift voucher with a follow-up questionnaire. METHODS: Participants in MINT were randomised to receive either: (a) a 5 POUNDSgift voucher (incentive group) or (b) no gift voucher (no incentive group), with their 4 month or 8 month follow-up questionnaire. We recorded, for each group, the number of questionnaires returned, the number returned without any chasing from the study office, the overall number of non responders (after all chasing efforts by the study office), and the costs of following up each group. RESULTS: 2144 participants were randomised, 1070 to the incentive group and 1074 to the no incentive group. The proportion of questionnaires returned (RR 1.10 (95% CI 1.05, 1.16)) and the proportion returned without chasing (RR 1.14 (95% CI 1.05, 1.24) were higher in the incentive group, and the overall non-response rate was lower (RR 0.68 (95% CI 0.53, 0.87)). Adjustment for injury severity and hospital of recruitment to MINT made no difference to these results, and there were no differences in results between the 4-month and 8-month follow up questionnaires. Analysis of costs suggested a cost of 67.29 POUNDS per additional questionnaire returned. CONCLUSION: Monetary incentives may be an effective way to increase the proportion of postal questionnaires returned and minimise loss to follow-up in clinical trials. TRIAL REGISTRATION NUMBER: ISRCTN61305297. PMID- 19545428 TI - A case for using grid architecture for state public health informatics: the Utah perspective. AB - This paper presents the rationale for designing and implementing the next generation of public health information systems using grid computing concepts and tools. Our attempt is to evaluate all grid types including data grids for sharing information and computational grids for accessing computational resources on demand. Public health is a broad domain that requires coordinated uses of disparate and heterogeneous information systems. System interoperability in public health is limited. The next-generation public health information systems must overcome barriers to integration and interoperability, leverage advances in information technology, address emerging requirements, and meet the needs of all stakeholders. Grid-based architecture provides one potential technical solution that deserves serious consideration. Within this context, we describe three discrete public health information system problems and the process by which the Utah Department of Health (UDOH) and the Department of Biomedical Informatics at the University of Utah in the United States has approached the exploration for eventual deployment of a Utah Public Health Informatics Grid. These three problems are: i) integration of internal and external data sources with analytic tools and computational resources; ii) provide external stakeholders with access to public health data and services; and, iii) access, integrate, and analyze internal data for the timely monitoring of population health status and health services. After one year of experience, we have successfully implemented federated queries across disparate administrative domains, and have identified challenges and potential solutions concerning the selection of candidate analytic grid services, data sharing concerns, security models, and strategies for reducing expertise required at a public health agency to implement a public health grid. PMID- 19545429 TI - Disease association with two Helicobacter pylori duplicate outer membrane protein genes, homB and homA. AB - BACKGROUND: homB encodes a Helicobacter pylori outer membrane protein. This gene was previously associated with peptic ulcer disease (PUD) and was shown to induce activation of interleukin-8 secretion in vitro, as well as contributing to bacterial adherence. Its 90%-similar gene, homA, was previously correlated with gastritis. The present study aimed to evaluate the gastric disease association with homB and homA, as well as with the H. pylori virulence factors cagA, babA and vacA, in 415 H. pylori strains isolated from patients from East Asian and Western countries. The correlation among these genotypes was also evaluated. RESULTS: Both homB and homA genes were heterogeneously distributed worldwide, with a marked difference between East Asian and Western strains. In Western strains (n = 234, 124 PUD and 110 non-ulcer dyspepsia (NUD), homB, cagA and vacA s1 were all significantly associated with PUD (p = 0.025, p = 0.014, p = 0.039, respectively), and homA was closely correlated with NUD (p = 0.072). In East Asian strains (n = 138, 73 PUD and 65 NUD), homB was found more frequently than homA, and none of these genes was associated with the clinical outcome. Overall, homB was associated with the presence of cagA (p = 0.043) and vacA s1 (p < 0.001), whereas homA was found more frequently in cagA-negative (p = 0.062) and vacA s2 (p < 0.001) strains. Polymorphisms in homB and homA copy number were observed, with a clear geographical specificity, suggesting an involvement of these genes in host adaptation. A correlation between the homB two-copy genotype and PUD was also observed, emphasizing the role of homB in the virulence of the strain. CONCLUSION: The global results suggest that homB and homA contribute to the determination of clinical outcome. PMID- 19545430 TI - The complexities of measuring access to parks and physical activity sites in New York City: a quantitative and qualitative approach. AB - BACKGROUND: Proximity to parks and physical activity sites has been linked to an increase in active behaviors, and positive impacts on health outcomes such as lower rates of cardiovascular disease, diabetes, and obesity. Since populations with a low socio-economic status as well as racial and ethnic minorities tend to experience worse health outcomes in the USA, access to parks and physical activity sites may be an environmental justice issue. Geographic Information systems were used to conduct quantitative and qualitative analyses of park accessibility in New York City, which included kernel density estimation, ordinary least squares (global) regression, geographically weighted (local) regression, and longitudinal case studies, consisting of field work and archival research. Accessibility was measured by both density of park acreage and density of physical activity sites. Independent variables included percent non-Hispanic black, percent Hispanic, percent below poverty, percent of adults without high school diploma, percent with limited English-speaking ability, and population density. RESULTS: The ordinary least squares linear regression found weak relationships in both the park acreage density and the physical activity site density models (R(a)(2) = .11 and .23, respectively; AIC = 7162 and 3529, respectively). Geographically weighted regression, however, suggested spatial non stationary in both models, indicating disparities in accessibility that vary over space with respect to magnitude and directionality of the relationships (AIC = 2014 and -1241, respectively). The qualitative analysis supported the findings of the local regression, confirming that although there is a geographically inequitable distribution of park space and physical activity sites, it is not globally predicted by race, ethnicity, or socio-economic status. CONCLUSION: The combination of quantitative and qualitative analyses demonstrated the complexity of the issues around racial and ethnic disparities in park access. They revealed trends that may not have been otherwise detectable, such as the spatially inconsistent relationship between physical activity site density and socio demographics. In order to establish a more stable global model, a number of additional factors, variables, and methods might be used to quantify park accessibility, such as network analysis of proximity, perception of accessibility and usability, and additional park quality characteristics. Accurate measurement of park accessibility can therefore be important in showing the links between opportunities for active behavior and beneficial health outcomes. PMID- 19545433 TI - Serum lipid profiles among patients initiating ritonavir-boosted atazanavir versus efavirenz-based regimens. AB - BACKGROUND: Antiretrovirals used to treat HIV-infected patients have the potential to adversely affect serum lipid profiles and increase the risk of cardiovascular disease which is an emerging concern among HIV-infected patients. Since boosted atazanavir and efavirenz are both considered preferred antiretrovirals a head to head comparison of their effects on serum lipids is needed. AIM: The primary objective of the study was to compare the effects of atazanavir (boosted and unboosted) and efavirenz based regimens on serum lipid profiles. STUDY DESIGN: Prospective cohort study nested within three ongoing cohorts of HIV-infected individuals. STUDY POPULATION AND METHODS: Participants initiating either atazanavir or efavirenz based regimens with documented pre- and post-initiation lipid values. Multivariate linear regression was conducted to estimate adjusted mean differences between treatment groups for high density lipoprotein cholesterol (HDL-c), non-HDL-c, and log total cholesterol (TC) to HDL c ratio outcomes; log-linear regression models were used to estimate differences in prevalence of low HDL-c and desirable TC. RESULTS: The final study population was comprised of 380 efavirenz and 281 atazanavir initiators. Both atazanavir and efavirenz users had increases in serum HDL-c and decreases in TC/HDL ratio. In comparison to individuals initiating efavirenz, boosted atazanavir users on average had lower HDL-c (-4.12 mg/dl, p < 0.001) and non HDL-c (-5.75 mg/dl, p < 0.01), but similar declines in TC/HDL ratio. CONCLUSION: Both efavirenz and atazanavir-based regimens (boosted and unboosted) resulted in similar beneficial declines in the TC/HDL ratio. PMID- 19545431 TI - Alveolar hypoxia, alveolar macrophages, and systemic inflammation. AB - Diseases featuring abnormally low alveolar PO2 are frequently accompanied by systemic effects. The common presence of an underlying inflammatory component suggests that inflammation may contribute to the pathogenesis of the systemic effects of alveolar hypoxia. While the role of alveolar macrophages in the immune and defense functions of the lung has been long known, recent evidence indicates that activation of alveolar macrophages causes inflammatory disturbances in the systemic microcirculation. The purpose of this review is to describe observations in experimental animals showing that alveolar macrophages initiate a systemic inflammatory response to alveolar hypoxia. Evidence obtained in intact animals and in primary cell cultures indicate that alveolar macrophages activated by hypoxia release a mediator(s) into the circulation. This mediator activates perivascular mast cells and initiates a widespread systemic inflammation. The inflammatory cascade includes activation of the local renin-angiotensin system and results in increased leukocyte-endothelial interactions in post-capillary venules, increased microvascular levels of reactive O2 species; and extravasation of albumin. Given the known extrapulmonary responses elicited by activation of alveolar macrophages, this novel phenomenon could contribute to some of the systemic effects of conditions featuring low alveolar PO2. PMID- 19545432 TI - In vivo proton magnetic resonance spectroscopy reveals region specific metabolic responses to SIV infection in the macaque brain. AB - BACKGROUND: In vivo proton magnetic resonance spectroscopy (1H-MRS) studies of HIV-infected humans have demonstrated significant metabolic abnormalities that vary by brain region, but the causes are poorly understood. Metabolic changes in the frontal cortex, basal ganglia and white matter in 18 SIV-infected macaques were investigated using MRS during the first month of infection. RESULTS: Changes in the N-acetylaspartate (NAA), choline (Cho), myo-inositol (MI), creatine (Cr) and glutamine/glutamate (Glx) resonances were quantified both in absolute terms and relative to the creatine resonance. Most abnormalities were observed at the time of peak viremia, 2 weeks post infection (wpi). At that time point, significant decreases in NAA and NAA/Cr, reflecting neuronal injury, were observed only in the frontal cortex. Cr was significantly elevated only in the white matter. Changes in Cho and Cho/Cr were similar across the brain regions, increasing at 2 wpi, and falling below baseline levels at 4 wpi. MI and MI/Cr levels were increased across all brain regions. CONCLUSION: These data best support the hypothesis that different brain regions have variable intrinsic vulnerabilities to neuronal injury caused by the AIDS virus. PMID- 19545434 TI - Conservation of the glucan phosphatase laforin is linked to rates of molecular evolution and the glucan metabolism of the organism. AB - BACKGROUND: Lafora disease (LD) is a fatal autosomal recessive neurodegenerative disease. A hallmark of LD is cytoplasmic accumulation of insoluble glucans, called Lafora bodies (LBs). Mutations in the gene encoding the phosphatase laforin account for approximately 50% of LD cases, and this gene is conserved in all vertebrates. We recently demonstrated that laforin is the founding member of a unique class of phosphatases that dephosphorylate glucans. RESULTS: Herein, we identify laforin orthologs in a protist and two invertebrate genomes, and report that laforin is absent in the vast majority of protozoan genomes and it is lacking in all other invertebrate genomes sequenced to date. We biochemically characterized recombinant proteins from the sea anemone Nematostella vectensis and the amphioxus Branchiostoma floridae to demonstrate that they are laforin orthologs. We demonstrate that the laforin gene has a unique evolutionary lineage; it is conserved in all vertebrates, a subclass of protists that metabolize insoluble glucans resembling LBs, and two invertebrates. We analyzed the intron-exon boundaries of the laforin genes in each organism and determine, based on recently published reports describing rates of molecular evolution in Branchiostoma and Nematostella, that the conservation of laforin is linked to the molecular rate of evolution and the glucan metabolism of an organism. CONCLUSION: Our results alter the existing view of glucan phosphorylation/dephosphorylation and strongly suggest that glucan phosphorylation is a multi-Kingdom regulatory mechanism, encompassing at least some invertebrates. These results establish boundaries concerning which organisms contain laforin. Laforin is conserved in all vertebrates, it has been lost in the vast majority of lower organisms, and yet it is an ancient gene that is conserved in a subset of protists and invertebrates that have undergone slower rates of molecular evolution and/or metabolize a carbohydrate similar to LBs. Thus, the laforin gene holds a unique place in evolutionary biology and has yielded insights into glucan metabolism and the molecular etiology of Lafora disease. PMID- 19545435 TI - Losartan counteracts the hyper-reactivity to angiotensin II and ROCK1 over activation in aortas isolated from streptozotocin-injected diabetic rats. AB - BACKGROUND: In streptozotocin-injected rats (STZ-rats), we previously demonstrated a role for angiotensin II (AT-II) in cardiac remodelling and insulin resistance partially counteracted by in vivo treatment with losartan, an AT-II receptor antagonist.We now aimed to investigate the effect of treating diabetic STZ-rats with losartan on diabetes vascular response to vasoconstrictors. METHODS: Male Wistar rats were randomly divided in four groups, two of them were assigned to receive losartan in the drinking water (20 mg/kg/day) until the experiment ending (3 weeks afterward). After 1 week, two groups, one of which receiving losartan, were injected in the tail vein with citrate buffer (normoglycemic, N and normoglycemic, losartan-treated, NL). The remaining received a single injection of streptozotocin (50 mg/kg in citrate i.v.) thus becoming diabetic (D) and diabetic losartan-treated (DL). Plasma glycaemia and blood pressure were measured in all animals before the sacrifice (15 days after diabetes induction).In aortic strips isolated from N, NL, D and DL rats we evaluated i) the isometric concentration-dependent contractile response to phenylephrine (Phe) and to AT-II; ii) the RhoA-kinase (ROCK1) activity and expression by enzyme-immunoassay and Western blot respectively. KEY RESULTS: The concentration-dependent contractile effect of Phe was similar in aortas from all groups, whereas at all concentrations tested, AT-II contraction efficacy was 2 and half and 1 and half times higher in D and DL respectively in comparison with N and NL. AT-II contracture was similarly reduced in all groups by AT-II receptor antagonists, irbesartan or irbesartan plus PD123319. HA-1077 (10 microM), an inhibitor of ROCK1 activity, reduced AT-II efficacy (Deltamg/mg tissue w.w.) by 3.5 +/- 1.0, -4.6 +/- 1.9, -22.1 +/- 2.2 and -11.4 +/- 1.3 in N, NL, D and DL respectively). ROCK1 activity and expression were higher in D than in N/NL and DL aortas. CONCLUSION AND IMPLICATIONS: Aortas isolated from STZ-rats present hyper contracture to AT-II mainly dependent on the up-regulation of ROCK1 expression/activity. In vivo losartan treatment partially corrects AT-II hyper contracture, limiting the increase in ROCK1 expression/activity. These data offer a new molecular mechanism supporting the rationale for using losartan in the prevention of diabetic vascular complications. PMID- 19545436 TI - Unsupervised assessment of microarray data quality using a Gaussian mixture model. AB - BACKGROUND: Quality assessment of microarray data is an important and often challenging aspect of gene expression analysis. This task frequently involves the examination of a variety of summary statistics and diagnostic plots. The interpretation of these diagnostics is often subjective, and generally requires careful expert scrutiny. RESULTS: We show how an unsupervised classification technique based on the Expectation-Maximization (EM) algorithm and the naive Bayes model can be used to automate microarray quality assessment. The method is flexible and can be easily adapted to accommodate alternate quality statistics and platforms. We evaluate our approach using Affymetrix 3' gene expression and exon arrays and compare the performance of this method to a similar supervised approach. CONCLUSION: This research illustrates the efficacy of an unsupervised classification approach for the purpose of automated microarray data quality assessment. Since our approach requires only unannotated training data, it is easy to customize and to keep up-to-date as technology evolves. In contrast to other "black box" classification systems, this method also allows for intuitive explanations. PMID- 19545437 TI - Collagen fleece-bound fibrin sealant is not associated with an increased risk of thromboembolic events or major bleeding after its use for haemostasis in surgery: a prospective multicentre surveillance study. AB - BACKGROUND: Topical haemostatic agents are used to help achieve haemostasis during surgery when standard surgical techniques are insufficient. The objective of this study was to confirm the safety profile of an equine collagen patch coated with human fibrinogen and human thrombin with particular focus on the occurrence of thromboembolic events (TEEs), major bleeding and immunological events. METHODS: This was a non-interventional, multicentre, prospective, surveillance study in which a collagen fleece-bound fibrin sealant was prescribed in accordance with its marketing authorisation. The decision to use the sealant was based solely on current surgical practice. All patients that received the sealant and provided informed consent were included. TEEs (any coagula-based occlusion in a vessel or the heart identified by symptomatic clinical signs and/or verified by paraclinical examination), major bleeding (any bleeding that required intervention), and immunological events (hypersensitivity including anaphylaxis) that occurred during surgery, post-operative hospital stay or 6 months of follow-up were reported as adverse events. The primary endpoint was the proportion of patients experiencing a confirmed TEE. RESULTS: A total of 3098 patients were recruited at 227 centres in 12 European countries. The most frequent types of surgery were hepatic (33%), gastrointestinal (16%) and urological (14%) and the main indication for surgery was for primary (35%) or secondary (20%) malignancy. Forty-six patients (1.5%, 95% CI 1.1-2.0%) had at least one TEE during the study. The most commonly reported TEEs were pulmonary embolism or post-procedural pulmonary embolism (n = 18) and deep vein thrombosis (n = 9). There were 64 major bleedings in 62 patients and 9 immunological events in 8 patients. CONCLUSION: Collagen fleece-bound fibrin sealant does not appear to be associated with an increased risk of TEEs, major bleeding or immunological events in patients undergoing surgery. TRIAL REGISTRATION: Clinicaltrials.gov number: NCT00285623. PMID- 19545439 TI - MicroCT for comparative morphology: simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues. AB - BACKGROUND: Comparative, functional, and developmental studies of animal morphology require accurate visualization of three-dimensional structures, but few widely applicable methods exist for non-destructive whole-volume imaging of animal tissues. Quantitative studies in particular require accurately aligned and calibrated volume images of animal structures. X-ray microtomography (microCT) has the potential to produce quantitative 3D images of small biological samples, but its widespread use for non-mineralized tissues has been limited by the low x ray contrast of soft tissues. Although osmium staining and a few other techniques have been used for contrast enhancement, generally useful methods for microCT imaging for comparative morphology are still lacking. RESULTS: Several very simple and versatile staining methods are presented for microCT imaging of animal soft tissues, along with advice on tissue fixation and sample preparation. The stains, based on inorganic iodine and phosphotungstic acid, are easier to handle and much less toxic than osmium, and they produce high-contrast x-ray images of a wide variety of soft tissues. The breadth of possible applications is illustrated with a few microCT images of model and non-model animals, including volume and section images of vertebrates, embryos, insects, and other invertebrates. Each image dataset contains x-ray absorbance values for every point in the imaged volume, and objects as small as individual muscle fibers and single blood cells can be resolved in their original locations and orientations within the sample. CONCLUSION: With very simple contrast staining, microCT imaging can produce quantitative, high-resolution, high-contrast volume images of animal soft tissues, without destroying the specimens and with possibilities of combining with other preparation and imaging methods. Such images are expected to be useful in comparative, developmental, functional, and quantitative studies of morphology. PMID- 19545438 TI - Non-Hodgkin lymphoma response evaluation with MRI texture classification. AB - BACKGROUND: To show magnetic resonance imaging (MRI) texture appearance change in non-Hodgkin lymphoma (NHL) during treatment with response controlled by quantitative volume analysis. METHODS: A total of 19 patients having NHL with an evaluable lymphoma lesion were scanned at three imaging timepoints with 1.5T device during clinical treatment evaluation. Texture characteristics of images were analyzed and classified with MaZda application and statistical tests. RESULTS: NHL tissue MRI texture imaged before treatment and under chemotherapy was classified within several subgroups, showing best discrimination with 96% correct classification in non-linear discriminant analysis of T2-weighted images.Texture parameters of MRI data were successfully tested with statistical tests to assess the impact of the separability of the parameters in evaluating chemotherapy response in lymphoma tissue. CONCLUSION: Texture characteristics of MRI data were classified successfully; this proved texture analysis to be potential quantitative means of representing lymphoma tissue changes during chemotherapy response monitoring. PMID- 19545441 TI - Family physicians' effort to stay in charge of the medical treatment when patients have home care by district nurses. A grounded theory study. AB - BACKGROUND: District nurses (DNs) provide home care for old persons with a mixture of chronic diseases, symptoms and reduced functional ability. Family physicians (FPs) have been criticised for their lack of involvement in this care. The aim of this study was to obtain increased knowledge concerning the FP's experience of providing medical treatment for patients with home care provided by DNs by developing a theoretical model that elucidates how FPs handle the problems they encounter regarding the individual patients and their conditions. METHODS: Semi-structured interviews were conducted with 13 Swedish FPs concerning one of their registered patients with home care by a DN, and the treatment of this patient. Grounded theory methodology (GTM) was used in the analyses. RESULTS: The core category was the effort to stay in charge of the medical treatment. This involved three types of problems: gaining sufficient insight, making adequate decisions, and maintaining appropriate medical treatment. For three categories of patients, the FPs had problems staying in charge. Patients with reduced functional ability had problems providing information and maintaining treatment. Patients who were "fixed in their ways" did not provide information and did not comply with recommendations, and for patients with complex conditions, making adequate decisions could be problematic. To overcome the problems, four different strategies were used: relying on information from others, supporting close observation and follow-up by others, being constantly ready to change the goal of the treatment, and relying on others to provide treatment. CONCLUSION: The patients in this study differed from most other patients seen at the healthcare centre as the consultation with the patient could not provide the usual foundation for decisions concerning medical treatment. Information from and collaboration with the DN and other home care providers was essential for the FP's effort to stay in charge of the medical treatment. The complexity of the situation made it problematic for the FP to make adequate decisions about the goal of the medical treatment. The goal of the treatment had to be constantly evaluated based on information from the DN and other care providers, and thus this information was absolutely crucial. PMID- 19545440 TI - Impairment of mitochondrial calcium handling in a mtSOD1 cell culture model of motoneuron disease. AB - BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by the selective loss of motor neurons (MN) in the brain stem and spinal cord. Intracellular disruptions of cytosolic and mitochondrial calcium have been associated with selective MN degeneration, but the underlying mechanisms are not well understood. The present evidence supports a hypothesis that mitochondria are a target of mutant SOD1-mediated toxicity in familial amyotrophic lateral sclerosis (fALS) and intracellular alterations of cytosolic and mitochondrial calcium might aggravate the course of this neurodegenerative disease. In this study, we used a fluorescence charged cool device (CCD) imaging system to separate and simultaneously monitor cytosolic and mitochondrial calcium concentrations in individual cells in an established cellular model of ALS. RESULTS: To gain insights into the molecular mechanisms of SOD1(G93A) associated motor neuron disease, we simultaneously monitored cytosolic and mitochondrial calcium concentrations in individual cells. Voltage - dependent cytosolic Ca2+ elevations and mitochondria - controlled calcium release mechanisms were monitored after loading cells with fluorescent dyes fura-2 and rhod-2. Interestingly, comparable voltage-dependent cytosolic Ca2+ elevations in WT (SH SY5Y(WT)) and G93A (SH-SY5Y(G93A)) expressing cells were observed. In contrast, mitochondrial intracellular Ca2+ release responses evoked by bath application of the mitochondrial toxin FCCP were significantly smaller in G93A expressing cells, suggesting impaired calcium stores. Pharmacological experiments further supported the concept that the presence of G93A severely disrupts mitochondrial Ca2+ regulation. CONCLUSION: In this study, by fluorescence measurement of cytosolic calcium and using simultaneous [Ca2+]i and [Ca2+]mito measurements, we are able to separate and simultaneously monitor cytosolic and mitochondrial calcium concentrations in individual cells an established cellular model of ALS. The primary goals of this paper are (1) method development, and (2) screening for deficits in mutant cells on the single cell level. On the technological level, our method promises to serve as a valuable tool to identify mitochondrial and Ca2+-related defects during G93A-mediated MN degeneration. In addition, our experiments support a model where a specialized interplay between cytosolic calcium profiles and mitochondrial mechanisms contribute to the selective degeneration of neurons in ALS. PMID- 19545443 TI - Delamination technique together with longitudinal incisions for treatment of Chiari I/syringomyelia complex: a prospective clinical study. AB - BACKGROUND: Treatment modalities in Chiari malformation type 1(CMI) accompanied by syringomyelia have not yet been standardized. Pathologies such as a small posterior fossa and thickened dura mater have been discussed previously. Various techniques have been explored to enlarge the foramen magnum and to expand the dura. The aim of this clinical study was to explore a new technique of excision of the external dura accompanied by widening the cisterna magna and making longitudinal incisions in the internal dura, without disturbing the arachnoid. METHODS: Ten patients with CMI and syringomyelia, operated between 2004 and 2006, formed this prospective series. All cases underwent foramen magnum decompression of 3 x 3 cm area with C1-C2 (partial) laminectomy, resection of foramen magnum fibrous band, excision of external dura, delamination and widening of internal dura with longitudinal incisions. RESULTS: Patients were aged between 25 and 58 years and occipital headache was the most common complaint. The mean duration of preoperative symptoms was 4 years and the follow-up time was 25 months. Clinical progression was halted for all patients; eight patients completely recovered and two reported no change. In one patient, there was a transient cerebrospinal fluid (CSF) fistula that was treated with tissue adhesive. While syringomyelia persisted radiologically with radiological stability in five patients; for three patients the syringomyelic cavity decreased in size, and for the remaining two it regressed completely. CONCLUSION: Removal of the fibrous band and the outer dural layer, at level of foramen magnum, together with the incision of inner dural layer appears to be good technique in adult CMI patients. The advantages are short operation time, no need for duraplasty, sufficient posterior fossa decompression, absence of CSF fistulas as a result of extra arachnoidal surgery, and short duration of hospitalization. Hence this surgical technique has advantages compared to other techniques. PMID- 19545442 TI - Marked differences in CRP genotype frequencies between the Fulani and sympatric ethnic groups in Africa. AB - BACKGROUND: C-reactive protein (CRP) is an acute phase protein that can activate various immune cells and bind to certain Fcgamma receptors. The latter may compete with the binding of IgG antibodies to these receptors and could thereby interfere with the antigen-specific immune response. Polymorphisms in the promoter region of the CRP gene have been strongly associated with the plasma concentration of CRP. The known lower susceptibility to malaria in the Fulani ethnic group, as compared to their sympatric neighbours in Africa, has been linked to different genetic backgrounds. The present study was performed to investigate if polymorphisms in the CRP gene could contribute to the lower susceptibility to malaria seen in the Fulani ethnic group. METHODS: The CRP -717 T>C, -286 C>T>A, and +1444 C>T polymorphisms were analysed in asymptomatic Fulani and non-Fulani individuals from Mali and Sudan using Pyrosequencing T and TaqMan r MGB probes. RESULTS: The rare -286 A allele, previously shown to be associated with increased CRP expression and plasma levels, was shown to be more frequent in the non-Fulani ethnic groups as compared to the sympatric Fulani ethnic group both in Mali and Sudan. The common -717 T allele was more prevalent in the non Fulani ethnic group compared to the sympatric Fulani ethnic group, but only in Mali. The parasite prevalence was increased for the -286 A allele, but not for the -717 T allele. No differences regarding genotype frequency or parasite prevalence were seen for +1444 C>T. CONCLUSION: This study indicate that CRP may play an important role in the immune responses to malaria, and that the -286 C/T/A CRP polymorphism may be a contributing factor to the lower susceptibility to malaria seen in the Fulani. PMID- 19545444 TI - The Ets dominant repressor En/Erm enhances intestinal epithelial tumorigenesis in ApcMin mice. AB - BACKGROUND: Ets transcription factors have been widely implicated in the control of tumorigenesis, with most studies suggesting tumor-promoting roles. However, few studies have examined Ets tumorigenesis-modifying functions in vivo using model genetic systems. METHODS: Using mice expressing a previously characterized Ets dominant repressor transgene in the intestinal epithelium (Villin-En/Erm), we examined the consequences of blocking endogenous Ets-mediated transcriptional activation on tumorigenesis in the ApcMin model of intestinal carcinoma. RESULTS: En/Erm expression in the intestine, at levels not associated with overt crypt villus dysmorphogenesis, results in a marked increase in tumor number in ApcMin animals. Moreover, when examined histologically, tumors from En/Erm-expressing animals show a trend toward greater stromal invasiveness. Detailed analysis of crypt-villus homeostasis in these En/Erm transgenic animals suggests increased epithelial turnover as one possible mechanism for the enhanced tumorigenesis. CONCLUSION: Our findings provide in vivo evidence for a tumor-restricting function of endogenous Ets factors in the intestinal epithelium. PMID- 19545445 TI - Rasch analysis of the Multiple Sclerosis Impact Scale MSIS-29. AB - BACKGROUND: Multiple Sclerosis (MS) is a degenerative neurological disease that causes impairments, including spasticity, pain, fatigue, and bladder dysfunction, which negatively impact on quality of life. The Multiple Sclerosis Impact Scale (MSIS-29) is a disease-specific health-related quality of life (HRQoL) instrument, developed using the patient's perspective on disease impact. It consists of two subscales assessing the physical (MSIS-29-PHYS) and psychological (MSIS-29-PSYCH) impact of MS. Although previous studies have found support for the psychometric properties of the MSIS-29 using traditional methods of scale evaluation, the scale has not been subjected to a detailed Rasch analysis. Therefore, the objective of this study was to use Rasch analysis to assess the internal validity of the scale, and its response format, item fit, targeting, internal consistency and dimensionality. METHODS: Ninety-two persons with definite MS residing in the community were recruited from a tertiary hospital database. Patients completed the MSIS-29 as part of a larger study. Rasch analysis was undertaken to assess the psychometric properties of the MSIS-29. RESULTS: Rasch analysis showed overall support for the psychometric properties of the two MSIS-29 subscales, however it was necessary to reduce the response format of the MSIS-29-PHYS to a 3-point response scale. Both subscales were unidimensional, had good internal consistency, and were free from item bias for sex and age. Dimensionality testing indicated it was not appropriate to combine the two subscales to form a total MSIS score. CONCLUSION: In this first study to use Rasch analysis to fully assess the psychometric properties of the MSIS-29 support was found for the two subscales but not for the use of the total scale. Further use of Rasch analysis on the MSIS-29 in larger and broader samples is recommended to confirm these findings. PMID- 19545446 TI - The TrueBlue study: is practice nurse-led collaborative care effective in the management of depression for patients with heart disease or diabetes? AB - BACKGROUND: In the presence of type 2 diabetes (T2DM) or coronary heart disease (CHD), depression is under diagnosed and under treated despite being associated with worse clinical outcomes. Our earlier pilot study demonstrated that it was feasible, acceptable and affordable for practice nurses to extend their role to include screening for and monitoring of depression alongside biological and lifestyle risk factors. The current study will compare the clinical outcomes of our model of practice nurse-led collaborative care with usual care for patients with depression and T2DM or CHD. METHODS: This is a cluster-randomised intervention trial. Eighteen general practices from regional and metropolitan areas agreed to join this study, and were allocated randomly to an intervention or control group. We aim to recruit 50 patients with co-morbid depression and diabetes or heart disease from each of these practices. In the intervention group, practice nurses (PNs) will be trained for their enhanced roles in this nurse-led collaborative care study. Patients will be invited to attend a practice nurse consultation every 3 months prior to seeing their usual general practitioner. The PN will assess psychological, physiological and lifestyle parameters then work with the patient to set management goals. The outcome of this assessment will form the basis of a GP Management Plan document. In the control group, the patients will continue to receive their usual care for the first six months of the study before the PNs undergo the training and switch to the intervention protocol. The primary clinical outcome will be a reduction in the depression score. The study will also measure the impact on physiological measures, quality of life and on patient attitude to health care delivered by practice nurses. CONCLUSION: The strength of this programme is that it provides a sustainable model of chronic disease management with monitoring and self management assistance for physiological, lifestyle and psychological risk factors for high-risk patients with co-morbid depression, diabetes or heart disease. The study will demonstrate whether nurse-led collaborative care achieves better outcomes than usual care. PMID- 19545447 TI - Ethnicity and OPRM variant independently predict pain perception and patient controlled analgesia usage for post-operative pain. AB - BACKGROUND: Morphine consumption can vary widely between individuals even for identical surgical procedures. As mu-opioid receptor (OPRM1) is known to modulate pain perception and mediate the analgesic effects of opioid compounds in the central nervous system, we examined the influence of two OPRM polymorphisms on acute post-operative pain and morphine usage in women undergoing elective caesarean delivery. RESULTS: Data on self-reported pain scores and amount of total morphine use according to patient-controlled analgesia were collected from 994 women from the three main ethnic groups in Singapore. We found statistically significant association of the OPRM 118A>G with self-administered morphine during the first 24-hour postoperative period both in terms of total morphine (p = 1.7 x 10(-5)) and weight-adjusted morphine (p = 6.6 x 10(-5)). There was also significant association of this OPRM variant and time-averaged self-rated pain scores (p = 0.024). OPRM 118G homozygotes used more morphine and reported higher pain scores than 118A carriers. Other factors which influenced pain score and morphine usage include ethnicity, age and paying class. CONCLUSION: Our results suggest that ethnicity and OPRM 118A>G genotype are independent and significant contributors to variation in pain perception and postoperative morphine use in patients undergoing cesarean delivery. PMID- 19545448 TI - A novel method, digital genome scanning detects KRAS gene amplification in gastric cancers: involvement of overexpressed wild-type KRAS in downstream signaling and cancer cell growth. AB - BACKGROUND: Gastric cancer is the third most common malignancy affecting the general population worldwide. Aberrant activation of KRAS is a key factor in the development of many types of tumor, however, oncogenic mutations of KRAS are infrequent in gastric cancer. We have developed a novel quantitative method of analysis of DNA copy number, termed digital genome scanning (DGS), which is based on the enumeration of short restriction fragments, and does not involve PCR or hybridization. In the current study, we used DGS to survey copy-number alterations in gastric cancer cells. METHODS: DGS of gastric cancer cell lines was performed using the sequences of 5000 to 15000 restriction fragments. We screened 20 gastric cancer cell lines and 86 primary gastric tumors for KRAS amplification by quantitative PCR, and investigated KRAS amplification at the DNA, mRNA and protein levels by mutational analysis, real-time PCR, immunoblot analysis, GTP-RAS pull-down assay and immunohistochemical analysis. The effect of KRAS knock-down on the activation of p44/42 MAP kinase and AKT and on cell growth were examined by immunoblot and colorimetric assay, respectively. RESULTS: DGS analysis of the HSC45 gastric cancer cell line revealed the amplification of a 500-kb region on chromosome 12p12.1, which contains the KRAS gene locus. Amplification of the KRAS locus was detected in 15% (3/20) of gastric cancer cell lines (8-18-fold amplification) and 4.7% (4/86) of primary gastric tumors (8-50 fold amplification). KRAS mutations were identified in two of the three cell lines in which KRAS was amplified, but were not detected in any of the primary tumors. Overexpression of KRAS protein correlated directly with increased KRAS copy number. The level of GTP-bound KRAS was elevated following serum stimulation in cells with amplified wild-type KRAS, but not in cells with amplified mutant KRAS. Knock-down of KRAS in gastric cancer cells that carried amplified wild-type KRAS resulted in the inhibition of cell growth and suppression of p44/42 MAP kinase and AKT activity. CONCLUSION: Our study highlights the utility of DGS for identification of copy-number alterations. Using DGS, we identified KRAS as a gene that is amplified in human gastric cancer. We demonstrated that gene amplification likely forms the molecular basis of overactivation of KRAS in gastric cancer. Additional studies using a larger cohort of gastric cancer specimens are required to determine the diagnostic and therapeutic implications of KRAS amplification and overexpression. PMID- 19545449 TI - Trends in esophageal cancer and body mass index by race and gender in the state of Michigan. AB - BACKGROUND: Adenocarcinoma of the esophagus has been increasing in incidence in the U.S. over the past several decades, particularly among white males. The factors driving the racial disparity in adenocarcinomas rates are not well understood. METHODS: Here we examine trends in both esophageal cancer incidence and body mass index (BMI) in a geographically defined cohort by gender and race. Age-adjusted esophageal cancer incidence rates from 1985 to 2005 were calculated from data collected by the Michigan state cancer registry. Trends were analyzed along with trends in BMI data obtained from the Behavioral Risk Factor Survey administered by the Centers for Disease Control. RESULTS: Overall, age adjusted incidence rates in esophageal carcinoma increased from 4.49 to 4.72 cases/100,000 persons per year in Michigan from 1985 to 2005. Among white males, the rate of adenocarcinomas increased by 0.21 cases/100,000 per year to a maximum of 6.40 cases/100,000 in 1999, after which these rates remained constant. There was a slight but non-significant increase in the rate of adenocarcinomas among African American males, for whom the average incidence rate was 8 times lower than that for white males (0.58 vs 4.72 cases/100,000 person years). While average BMI is rising in Michigan (from 26.68 in 1988 to 30.33 in 2005), average BMI was slightly higher among African Americans on average, and the rates of increase in BMI were not different between African American males and white males. CONCLUSION: The disparity between African American males and white males is not explained by ecological-level trends in BMI. Further research to identify the factors responsible for this disparity, possibly including anatomic fat distribution, are required. PMID- 19545451 TI - Combined effects of obesity and type 2 diabetes contribute to increased breast cancer risk in premenopausal women. AB - BACKGROUND: Both obesity and type 2 diabetes are among the risk factors for breast cancer development. Combined effect of these metabolic abnormalities on breast cancer risk however, has not been examined in premenopausal women. We tested this association in type 2 diabetic women, categorized as obese, overweight and normal body weight groups based on BMI. DESIGN AND METHODS: A total of 101 subjects were included in this study. Serum levels of IL-6, TNF alpha, C reactive protein, leptin, TGF-alpha, adiponectin and insulin were measured by ELISA. Data were logarithmically transformed for variables not normally distributed. Analysis of variance with post-hoc Bonferroni was applied to compare the data between the groups. Simple and partial correlation coefficients between the variables were determined and a stepwise multiple linear regression analysis was performed to determine the relationships between the variables of interest. RESULTS: Significantly increased levels of IL-6, C reactive protein, leptin and significantly decreased levels of adiponectin were found in obese group, while the levels of TNF-alpha and TGF-alpha were unaltered. A positive correlation between waist circumference and IL-6 was found in obese group. Similarly, C reactive protein, waist and hip circumferences were linearly correlated with BMI in obese group. Stepwise multiple linear regression analysis revealed several significant predictors for breast cancer risk. CONCLUSION: Obesity and type 2 diabetes, owing to their effects on adipocytokines and inflammatory mediators, contribute to increased breast cancer risk in premenopausal women. This study emphasizes healthy life style and better management of these metabolic disorders to avoid the pathogenesis of breast cancer and of other chronic diseases. PMID- 19545450 TI - Subcellular localization of the antidepressant-sensitive norepinephrine transporter. AB - BACKGROUND: Reuptake of synaptic norepinephrine (NE) via the antidepressant sensitive NE transporter (NET) supports efficient noradrenergic signaling and presynaptic NE homeostasis. Limited, and somewhat contradictory, information currently describes the axonal transport and localization of NET in neurons. RESULTS: We elucidate NET localization in brain and superior cervical ganglion (SCG) neurons, aided by a new NET monoclonal antibody, subcellular immunoisolation techniques and quantitative immunofluorescence approaches. We present evidence that axonal NET extensively colocalizes with syntaxin 1A, and to a limited degree with SCAMP2 and synaptophysin. Intracellular NET in SCG axons and boutons also quantitatively segregates from the vesicular monoamine transporter 2 (VMAT2), findings corroborated by organelle isolation studies. At the surface of SCG boutons, NET resides in both lipid raft and non-lipid raft subdomains and colocalizes with syntaxin 1A. CONCLUSION: Our findings support the hypothesis that SCG NET is segregated prior to transport from the cell body from proteins comprising large dense core vesicles. Once localized to presynaptic boutons, NET does not recycle via VMAT2-positive, small dense core vesicles. Finally, once NET reaches presynaptic plasma membranes, the transporter localizes to syntaxin 1A-rich plasma membrane domains, with a portion found in cholera toxin-demarcated lipid rafts. Our findings indicate that activity-dependent insertion of NET into the SCG plasma membrane derives from vesicles distinct from those that deliver NE. Moreover, NET is localized in presynaptic membranes in a manner that can take advantage of regulatory processes targeting lipid raft subdomains. PMID- 19545452 TI - Are measurements of systolic myocardial velocities and displacement with colour and spectral Tissue Doppler compatible? AB - BACKGROUND: Tissue Doppler (TD) in pulsed mode (spectral TD) and colour TD are the two modalities today available in tissue velocity echocardiography (TVE). Previous studies have shown poor agreement between these two methods when measuring myocardial velocities and displacement. In this study, the concordance between the myocardial velocity and displacement measurements using colour TD and different spectral TD procedures was evaluated. METHODS: Left ventricular (LV) longitudinal systolic myocardial velocities and displacement during ejection period were quantified at the basal septal and lateral wall in 24 healthy individuals (4 women and 20 men, 34 +/- 12 years) using spectral TD, colour TD and M-mode recordings. Mean, maximal and minimal spectral TD systolic velocities and the corresponding displacement values were obtained by measurements at the outer and inner borders of the spectral velocity signal. The results were then compared with those obtained with the two other modalities used. RESULTS: Systolic myocardial velocities derived from mean spectral TD frequencies were highly concordant with corresponding colour TD measurements (mean difference 0.10 +/- 0.54 cm/sec in septal and 0.09 +/- 0.97 cm/sec in lateral wall). Similarly, the agreement between spectral and colour TD (mean difference 0.22 +/- 0.74 mm in septal and 0.02 +/- 0.86 mm in lateral wall) as well as M-mode was good when mean spectral velocities were temporally integrated and the results did not differ statistically. Conversely, displacement values from the inner or outer border of the spectral signal differed significantly from values obtained with colour TD and M-mode (p < 0.001, in both cases). CONCLUSION: LV systolic myocardial measurements based on mean spectral TD frequencies are highly concordant with those provided by colour TD and M-mode. Hence, in order to maintain compatibility of the results, the use of this particular spectral TD procedure should be advocated in clinical praxis. PMID- 19545455 TI - Women's health, breast health: a review of the gynecologic effects of breast cancer. AB - Breast cancer is very common and seen in both premenopausal and postmenopausal women. Research into prevention, better screening, and more effective treatments is occurring continually, and changes are translated into clinical practice relatively quickly. It is important for women's health care providers to have an understanding of breast cancer treatments and the gynecologic side effects. For premenopausal women interested in fertility, options should be discussed prior to chemotherapy. Issues pertaining to pregnancy after breast cancer should be discussed in a multidisciplinary fashion, involving the obstetrician/gynecologist, breast surgeon, and oncologist. Ovarian suppression is often used as part of breast cancer treatment in premenopausal women with hormone positive disease, and menopausal symptoms may be severe. Hormonal therapies including tamoxifen and the aromatase inhibitors are used in the treatment of hormone positive breast cancers. Each of these drugs has a variety of gynecologic implications. Understanding the options for treatment for menopausal complaints in breast cancer patients is important for women's health providers. Although most breast cancers are sporadic, a small percentage will be due to mutations in the BRCA genes. It is important for women's health providers to take an appropriate family history and refer to genetic counselors for possible testing when hereditary cancer is suspected. This review focuses on the various women's health issues pertaining to breast cancer and treatment. PMID- 19545454 TI - Intraoperative ultrasound-guided iodine-125 seed implantation for unresectable pancreatic carcinoma. AB - BACKGROUND: To assess the feasibility and efficacy of using 125I seed implantation under intraoperative ultrasound guidance for unresectable pancreatic carcinoma. METHODS: Fourteen patients with pancreatic carcinoma that underwent laparotomy and considered unresectable were included in this study. Nine patients were pathologically diagnosed with Stage II disease, five patients with Stage III disease. Fourteen patients were treated with 125I seed implantation guided by intraoperative ultrasound and received D90 of 125I seeds ranging from 60 to 140 Gy with a median of 120 Gy. Five patients received an additional 35-50 Gy from external beam radiotherapy after seed implantation and six patients received 2-6 cycles of chemotherapy. RESULTS: 87.5% (7/8) of patients received partial to complete pain relief. The response rate of tumor was 78.6%, One-, two-and three year survival rates were 33.9% and 16.9%, 7.8%, with local control of disease achieved in 78.6% (11/14), and the median survival was 10 months (95% CI: 7.7 12.3). CONCLUSION: There were no deaths related to 125I seed implant. In this preliminary investigation, 125I seed implant provided excellent palliation of pain relief, local control and prolong the survival of patients with stage II and III disease to some extent. PMID- 19545453 TI - Preventing knee injuries in adolescent female football players - design of a cluster randomized controlled trial [NCT00894595]. AB - BACKGROUND: Knee injuries in football are common regardless of age, gender or playing level, but adolescent females seem to have the highest risk. The consequences after severe knee injury, for example anterior cruciate ligament (ACL) injury, are well-known, but less is known about knee injury prevention. We have designed a cluster randomized controlled trial (RCT) to evaluate the effect of a warm-up program aimed at preventing acute knee injury in adolescent female football. METHODS: In this cluster randomized trial 516 teams (309 clusters) in eight regional football districts in Sweden with female players aged 13-17 years were randomized into an intervention group (260 teams) or a control group (256 teams). The teams in the intervention group were instructed to do a structured warm-up program at two training sessions per week throughout the 2009 competitive season (April to October) and those in the control group were informed to train and play as usual. Sixty-eight sports physical therapists are assigned to the clubs to assist both groups in data collection and to examine the players' acute knee injuries during the study period. Three different forms are used in the trial: (1) baseline player data form collected at the start of the trial, (2) computer-based registration form collected every month, on which one of the coaches/team leaders documents individual player exposure, and (3) injury report form on which the study therapists report acute knee injuries resulting in time loss from training or match play. The primary outcome is the incidence of ACL injury and the secondary outcomes are the incidence of any acute knee injury (except contusion) and incidence of severe knee injury (defined as injury resulting in absence of more than 4 weeks). Outcome measures are assessed after the end of the 2009 season. DISCUSSION: Prevention of knee injury is beneficial for players, clubs, insurance companies, and society. If the warm-up program is proven to be effective in reducing the incidence of knee injury, it can have a major impact by reducing the future knee injury burden in female football as well as the negative long-term disabilities associated with knee injury. TRIAL REGISTRATION: NCT00894595. PMID- 19545457 TI - Factors affecting maternal perception of fetal movement. AB - Maternal perception of fetal movement is an important screening method for fetal well-being, as decreased fetal movement is associated with a range of pregnancy pathologies and poor pregnancy outcomes. An understanding of factors that may affect perception could help clinicians to determine the importance of maternal reports of decreased fetal movement. This review considers factors that may affect maternal perception of fetal movement and the sensitivity of maternal perception of fetal movements in comparison with ultrasound and other objective methods of movement detection. There is conflicting evidence on whether parity, gestational age, overweight and obesity, and placental location affect perception. This may be related to the small sample sizes of available studies and lack of consistent definitions of factors that may affect the ability of mothers to perceive movement. There is some evidence that psychological factors and duration of fetal movement may affect perception, and that strong movements and those including trunk movement are more likely to be perceived. The proportion of fetal breathing movements that mothers perceive has not been investigated. Research is also lacking as to whether there needs to be contact of fetal part(s) with maternal structures for movement to be perceived, and whether fetal position, amniotic fluid volume, maternal position, sedatives, or other drugs affect movement perception. PMID- 19545456 TI - Diagnosis of acute appendicitis during pregnancy: a systematic review. AB - OBJECTIVE: To perform a systematic review to evaluate the diagnostic performance of computed tomography (CT) and magnetic resonance imaging (MRI) for the diagnosis of appendicitis in pregnancy. DATA SOURCES: A systematic literature search of MEDLINE from 1966 through August 2008, MEDION database, OVID MEDLINE from 1950 through August 2008, and bibliographies of review articles and eligible studies. METHODS OF STUDY SELECTION: Three articles related to the use of CT and 5 to the use of MRI for the diagnosis of acute appendicitis during pregnancy were identified. All of the identified studies were retrospective. Findings were compared to surgical pathology and/or clinical follow-up. Results were pooled using the Mantel-Haenszel fixed-effects model and the DerSimonian-Laird random effects model. TABULATION, INTEGRATION, AND RESULTS: The pooled estimates of sensitivity and specificity, positive and negative likelihood ratios for the performance of CT in patients with prior normal/inconclusive ultrasonography result were 85.7% (95% CI: 63.7%-97%) and 97.4% (95% CI: 86.2%-99.9%), 10.1 (95% CI: 3.4-30.1), and 0.21 (95% CI: 0.05-0.88), respectively. The pooled estimates of sensitivity and specificity, positive and negative likelihood ratios for performance of MRI in patients with prior normal/inconclusive ultrasonography result were 80% (95% CI: 44%-98%) and 99% (95% CI: 94%-100%), 22.7 (95% CI: 6.0 87.5), and 0.29 (95% CI: 0.13-0.68), respectively. CONCLUSION: This review is limited by the small number and retrospective nature of the available studies. With these limitations in mind, CT and MRI seem to be highly sensitive and specific for the diagnosis of appendicitis in pregnancy and their use should be considered when the results of ultrasonography are normal or inconclusive and appendicitis is suspected. PMID- 19545458 TI - Oral supplementation of propionyl-l-carnitine reduces body weight and hyperinsulinaemia in obese Zucker rats. AB - Propionyl-L-carnitine (PLC) is an SCFA esterified to carnitine that plays an important role in fatty acid oxidation and energy expenditure, in addition to having a protective effect on the endothelium. In order to evaluate the effect of PLC on an animal model of obesity, insulin resistance and, consequently, endothelial dysfunction, lean and obese Zucker rats (OZR) received either vehicle or PLC-supplemented drinking water (200 mg/kg per d) for 20 weeks. Body weight, food intake, systolic blood pressure and heart rate were controlled weekly and an oral glucose tolerance test was performed. Fasting glucose, TAG, cholesterol, HDL, NEFA, adiponectin and insulin were analysed in serum. Visceral adipose tissue and liver were weighed and liver TAG liver composition was evaluated. Endothelial and vascular functions were assessed in the aorta and small mesenteric arteries by response to acetylcholine, sodium nitroprusside and phenylephrine (Phe); NO participation was evaluated after incubation with the NO synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester (L-NAME) and endothelial NOS protein expression by Western blotting. PLC decreased body-weight gain, food intake, adiposity, insulin serum concentration and TAG liver content and improved insulin resistance. Aortae from OZR receiving either vehicle or PLC exhibited a lower contractile response to Phe. PLC-treated OZR showed an enhanced release of endothelial NO upon the adrenergic stimulation. The protection of vascular function found after treatment with PLC in an animal model of insulin resistance supports the necessity of clinical trials showing the effect of L carnitine supplements on metabolic disorders. PMID- 19545459 TI - The needle and the damage done: pericardial effusion with tamponade after needle ingestion in an infant. AB - OBJECTIVE: Paediatric foreign bodies may present with vague and nonspecific symptoms. It is important to have a high index of suspicion when managing such cases. METHOD: We report the case of a nine-month-old infant who presented with a wheeze, cough and fever following ingestion of a needle. RESULTS: This patient developed pericardial tamponade as a consequence of the needle ingestion, and required a thoracotomy for retrieval. We discuss the pathophysiology involved and the surgery required. CONCLUSION: Pericardial tamponade is a rare but potentially fatal manifestation of an ingested foreign body. PMID- 19545460 TI - Lymphangioma circumscriptum of the tongue: successful treatment using intralesional bleomycin. AB - OBJECTIVE: We report a case of lymphangioma circumscriptum of the tongue, a very rare site of occurrence, which was successfully managed with intralesional bleomycin therapy. METHOD: We present a case report and review of available literature regarding lymphangioma circumscriptum of the tongue and the role of bleomycin therapy. RESULTS: The patient was a 19-year-old man with a long standing lesion involving the tongue, who presented with spontaneous, episodic bleeding over the previous few months. A diagnosis of lymphangioma circumscriptum was established on biopsy. Intralesional bleomycin injection resulted in successful resolution, and the patient remained asymptomatic over more than one year's follow up. CONCLUSION: Lymphangioma circumscriptum is usually seen in the extremities and genitals. This case had a very rare site of occurrence, the tongue, and was successfully managed with conservative treatment, using intralesional bleomycin alone. PMID- 19545461 TI - Gustatory otorrhoea: a rare case of congenital external ear salivary fistula. AB - OBJECTIVE: We present an extremely rare case of a 44-year-old woman with right gustatory otorrhoea and otalgia. CASE REPORT: The patient had been initially treated for otitis externa after Pseudomonas aeruginosa was grown from a microbiological swab. The otorrhoea fluid was collected and tested positive for amylase. Sialography and computed tomography imaging of the temporal bone confirmed a sialo-aural fistula from the right parotid gland to the bony external acoustic meatus. The defect was consistent with a patent foramen of Huschke. The fistula was identified surgically via a superficial parotidectomy approach, after contrast injection of Bonney's blue dye into the parotid duct, and then ligated and divided. The patient had immediate and sustained resolution of her otorrhoea. CONCLUSIONS: Sialo-aural fistulae are extremely rare, and usually arise as a complication of surgery or as an acquired disease process. To date, only four cases have been reported. This case demonstrates the use of sensitive investigation involving sialography and computed tomography, as well as successful surgical management, with complete resolution of symptoms. PMID- 19545462 TI - Organised haematoma of the sphenoid sinus mimicking a pituitary tumour. AB - OBJECTIVE: We report an extremely rare case of an organised haematoma arising in the sphenoid sinus. CASE REPORT: An 85-year-old woman presented with an expansile soft tissue mass in the left sphenoid sinus, with bony destruction of the sella turcica, which mimicked the extrasellar extension of a pituitary tumour. The tumour was excised using an endoscopic, transsphenoidal approach. Histopathological examination revealed an organised haematoma. CONCLUSION: To our knowledge, this is the first report of an organised haematoma arising in the sphenoid sinus. This case indicates that organised haematoma should be included in the differential diagnosis of extensive sphenoid lesions; it also emphasises the importance of correct pre-operative diagnosis for therapeutic planning, as complete surgical resection by an endoscopic approach is curative. PMID- 19545463 TI - Prevention of blood staining of endoscope tip during Functional Endoscopic Sinus Surgery: sleeve technique. AB - Optimisation of the endoscopic view during FESS may require correction of a deviated septum. The resulting incision leads to bleeding which could obscure the view of the endoscope. Repeated cleaning of the endoscope intraoperatively is time consuming and traumatisation of the lining of the nasal mucosa may lead to formation of adhesions post operatively. We discuss the use a segment of suction tubing that can act as a conduit or sleeve for the passage of the endoscope into the nasal cavity. This protects the endoscope tip from the bleeding area. PMID- 19545464 TI - Two siblings with progressive, fluctuating hearing loss after head trauma, treated with cochlear implantation. AB - OBJECTIVE: Description of two siblings with unexplained, progressive, perceptive hearing loss after head trauma. DESIGN: Case report. SUBJECTS: Two siblings aged six and eight years old with bilateral, intermittent but progressive hearing loss. RESULTS: These patients had a c.1172G>A (p.Ser391Asn) mutation in the SLC26A4 gene, which has not previously been reported and which caused Pendred or enlarged vestibular aqueduct syndrome. The diagnosis was based on the perceptive hearing loss, computed tomography findings and mutation analysis. The patients were each fitted with a cochlear implant because of their severe, progressive, perceptive hearing loss with deep fluctuations. The results were good. CONCLUSION: Further testing for the presence of an enlarged vestibular aqueduct is recommended when children present with sudden progression in perceptive hearing loss, whether or not in combination with head trauma. Cochlear implantation is indicated in patients with persistent, progressive hearing loss that leads to deafness. Implantation can be undertaken successfully despite cochlear hypoplasia. PMID- 19545465 TI - Successful use of intravenous amiodarone in a child with combined postoperative junctional and ectopic tachycardias. AB - Early postoperative arrhythmias are a known complication of cardiac surgery. It is unusual, however, to encounter postoperative junctional and ectopic atrial tachycardias in the same patient. We describe our experience with a 2-year-old girl who suffered both these tachycardias after repair of a ventricular septal defect, the abnormal rhythms being controlled solely with intravenous administration of amiodarone. PMID- 19545466 TI - BMI centile curves for Japanese children aged 5-17 years in 2000-2005. AB - OBJECTIVES: To examine changes in the age-specific prevalence of overweight and obesity in Japanese children between 2000 and 2005 and compare Japanese centile curves with international ones. DESIGN: A large, Japanese, representative cross sectional growth study. SETTING: Japan. SUBJECTS: Japanese children aged 5-17 years, including 669,986 subjects in 2000, 670,143 in 2001, 668,760 in 2002, 670,785 in 2003, 669,120 in 2004 and 668,234 in 2005. MAIN OUTCOME MEASURE: BMI (weight/height2). RESULTS: The prevalence of overweight and obesity in children aged 5-17 years decreased from 2000 to 2005 in both males and females, according to the definitions of the Centers for Disease Control and Prevention and the International Obesity Taskforce. For each year, the centile curves were drawn so that at age 17 years they passed through respectively 24.46 kg/m2 and 24.70 kg/m2 for males, and 29.41 kg/m2 and 29.69 kg/m2 for females, which are the BMI values at age 17 years from the international reference curves. CONCLUSIONS: The study results indicate that centile curves can change according to the time of observation, thus suggesting that international reference curves therefore need to be regularly revised while including more data from a larger range of countries. PMID- 19545467 TI - Overweight in dogs, but not in cats, is related to overweight in their owners. AB - OBJECTIVE: To quantify the environmental component of aetiology of overweight and obesity by examining the relationship between the degree of overweight in dogs and cats and the degree of overweight in their owners. DESIGN: Cross-sectional study. Main outcome measures of the owners were weight, height (stature) and BMI. Of the animals, weight and divergence from ideal weight were measured by a veterinarian. SETTING: Three veterinary clinics in Amsterdam, The Netherlands. SUBJECTS: Dogs and cats, together with their owners, who visited the veterinary clinic. Dogs and cats had to be older than 1 year, and their owners had to be at least 21 years old. After exclusion, there remained forty-seven pairs of dogs and their owners and thirty-six pairs of cats and their owners. RESULTS: We found a significant relationship between the degree of overweight of dogs and the BMI of their owners (r = 0.31). Correction for length of ownership, gender and age of the animal, and gender, age, education level and activity score of the owner did not materially affect this relationship. However, after correction for the amount of time the dog was being walked each day, this relationship disappeared. No significant relationship was found between the degree of overweight of cats and the BMI of their owners. CONCLUSIONS: The degree to which dogs are overweight is, in contrast to the degree to which cats are overweight, related to the BMI of their owners. PMID- 19545468 TI - Diet quality of a population sample from coastal north-east Spain evaluated by a Mediterranean adaptation of the diet quality index (DQI). AB - OBJECTIVE: To assess the adherence to the Mediterranean dietary pattern in the population from a coastal region from north-east Spain and its relationship to diseases, applying the Mediterranean Diet Quality Index (M-DQI) validated by the use of several biomarkers. DESIGN: Cross-sectional nutrition survey. SETTING: Population-based random sample derived from the Catalan Nutrition Survey. SUBJECTS: A total of 621 healthy adults. RESULTS: The Catalan representative sample presented a mean M-DQI score of 6.6 (sd 2.3, median 7, range 0-14). The percentage of adherence to the Mediterranean diet was 53 %; 10 % of subjects showed high adherence to the Mediterranean diet, while only 2 % were categorized as poorest adherence. The plasma fatty acid profile of the Catalan sample progressed with perfect regularity throughout the index ranges. Both EPA and DHA presented a significant correlation to the M-DQI (r = -0.410 for EPA and -0.360 for DHA). A significant increase in palmitic, oleic and alpha-linolenic acids and a significant decrease in stearic, linoleic and arachidonic acids content were also observed. The mean values for the M-DQI according to the clinical characteristics of the Catalan sample were also calculated. CONCLUSIONS: The M DQI has been demonstrated a suitable tool for assessment of an individual's nutritional status according to the Mediterranean dietary pattern and for clinical purposes. Although the current diet followed in Catalonia seems to agree with the main characteristics of the Mediterranean diet, the promotion of the Mediterranean pattern should be reinforced in the Catalan population, especially among young people. PMID- 19545469 TI - Nutrition and health education intervention for student volunteers: topic-wise assessment of impact using a non-parametric test. AB - OBJECTIVE: To assess the impact of a classroom-based nutrition and health education intervention among student community volunteers in improving their knowledge on individual topics. DESIGN: Prospective follow-up study. Topic-wise knowledge change among student volunteers on individual topics (twenty-one questions related to nutrition and health, eight questions related to infectious diseases and two questions related to obesity and hypertension) pertaining to nutrition and health was evaluated at baseline and after intervention, using the McNemar test. SETTING: Six different colleges affiliated to Osmania University, Andhra Pradesh, India. SUBJECTS: Six hundred and eighty-seven student volunteers under the National Service Scheme, of both genders, average age 19 years. RESULTS: A significant mean improvement of 11.36 (sd 8.49, P < 0.001) was observed in the overall nutrition and health knowledge scores of the student volunteers after the education intervention. The McNemar test showed that knowledge on individual topics related to energy, proteins, fats, adolescent phase, obesity, some lifestyle diseases and infectious diseases improved significantly (P < 0.01). No significant (P > 0.05) improvement was observed in knowledge on the nutritional content of milk and sprouted grams, hypertension, HIV/AIDS, ELISA and malaria. CONCLUSIONS: Topics on which our educational intervention could not bring about significant knowledge improvement have been identified and suitable modifications can be carried out to strengthen them. PMID- 19545470 TI - The commercial marketing of healthy lifestyles to address the global child and adolescent obesity pandemic: prospects, pitfalls and priorities. AB - Public- and private-sector initiatives to promote healthy eating and physical activity, called 'healthy lifestyles', are a relatively recent response to the global obesity pandemic. The present paper explores different views about marketing healthy lifestyles with a special emphasis on private-sector initiatives and public-private partnerships designed to reach young people. We discuss aspects of these initiatives and partnerships from three perspectives: (i) the potential for commercial marketing practices to have a favourable influence on reversing global obesity trends (termed prospects); (ii) unresolved dilemmas and challenges that may hinder progress (termed pitfalls); and (iii) the implementation and evaluation of coordinated and systematic actions (termed priorities) that may increase the likelihood that commercially marketed healthy lifestyle initiatives and public-private partnerships can make a positive contribution to reverse the rise in overweight and obesity among young people globally. PMID- 19545471 TI - Diet quality, nutrition and physical activity among adolescents: the Web-SPAN (Web-Survey of Physical Activity and Nutrition) project. AB - OBJECTIVE: To assess the overall diet quality of a sample of adolescents living in Alberta, Canada, and evaluate whether diet quality, nutrient intakes, meal behaviours (i.e. meal skipping and consuming meals away from home) and physical activity are related. DESIGN: A cross-sectional study design. Students completed the self-administered Web-Survey of Physical Activity and Nutrition (Web-SPAN). Students were classified as having poor, average or superior diet quality based on Canada's Food Guide to Healthy Eating (CFGHE). SETTING: One hundred and thirty six schools (37 %) within forty-four public and private school boards (75 %) in Alberta, Canada. SUBJECTS: Grade 7 to 10 Alberta students (n 4936) participated in the school-based research. RESULTS: On average, students met macronutrient requirements; however, micronutrient and fibre intakes were suboptimal. Median CFGHE food group intakes were below recommendations. Those with poor diet quality (42 %) had lower intakes of protein, fibre and low-calorie beverages; higher intakes of carbohydrates, fat and Other Foods (e.g. foods containing mostly sugar, high-salt/fat foods, high-calorie beverages, low-calorie beverages and high-sugar/fat foods); a lower frequency of consuming breakfast and a higher frequency of consuming meals away from home; and a lower level of physical activity when compared with students with either average or superior diet quality. CONCLUSIONS: Alberta adolescents were not meeting minimum CFGHE recommendations, and thus had suboptimal intakes and poor diet quality. Suboptimal nutritional intakes, meal behaviours and physical inactivity were all related to poor diet quality and reflect the need to target these health behaviours in order to improve diet quality and overall health and wellness. PMID- 19545472 TI - An investigation of the ways in which public health nutrition policy and practices can address climate change. AB - OBJECTIVE: To develop a framework to guide action in the public health nutrition workforce to develop policies and practices addressing factors contributing to climate change. DESIGN: Action/consultative research. SETTING: Interviews - South Australia, questionnaire - Australia. SUBJECTS: Interviews - key informants (n 6) were from various government, academic and non-government positions, invited through email. Questionnaire - participants were members of the public health nutrition workforce (n 186), recruited to the study through emails from public health nutrition contacts for each State in Australia (with the exception of South Australia). RESULTS: Support by participants for climate change as a valid role for dietitians and nutritionists was high (78 %). However, climate change was ranked low against other public health nutrition priorities. Support of participants to conduct programmes to address climate change from professional and work organisations was low. The final framework developed included elements of advocacy/lobbying, policy, professional recognition/support, organisational support, knowledge/skills, partnerships and programmes. CONCLUSIONS: This research demonstrates a need for public health nutrition to address climate change, which requires support by organisations, policy, improved knowledge and increased professional development opportunities. PMID- 19545473 TI - Factors associated with overweight in children in Rasht, Iran: gender, maternal education, skipping breakfast and parental obesity. AB - OBJECTIVE: The present study aimed to investigate the determinants of overweight and obesity among 6- to 11-year-old schoolchildren in Rasht, Iran. DESIGN: Cross sectional survey. Data on age, weekly frequency of skipping breakfast, physical activity and hours of television viewing were collected. Information on birth weight, parental age, parental educational levels, parental weight and height, and mother's employment status were gathered through self-administrated questionnaires given to the parents. SETTING: Elementary schools in Rasht. SUBJECTS: A total of 6635 children (3551 boys and 3084 girls) attending elementary schools in Rasht were studied. RESULTS: The overall prevalence of overweight was 11.5 % and 15.0 % for boys and girls, respectively; while the overall prevalence of obesity was 5.0 % and 5.9 %, respectively. Children with more educated mothers had a higher prevalence of overweight than children with less educated mothers. Logistic regression analysis showed that children with overweight/obese parents, children with more educated mothers and children who often skipped breakfast were more prone to overweight and obesity. CONCLUSIONS: These data suggest that overweight and obesity is a public health concern in this age group in Rasht. The observed sex and social differences in the prevalence of overweight and obesity call for policy makers' attention. PMID- 19545474 TI - Influence of acute caffeine on 50-kHz ultrasonic vocalizations in male adult rats and relevance to caffeine-mediated psychopharmacological effects. AB - To further characterize caffeine-mediated psychopharmacological effects, the present study investigated whether acute caffeine (3, 10, 30, 50 mg/kg i.p.) exerted any influence on the emission and features of ultrasonic vocalizations (USVs), which are thought to index changes involving emotional state, in male adult rats. The results obtained demonstrate that caffeine can trigger modifications in the maximum peak frequency and bandwidth of the 50-kHz range USVs. However, such an effect was not accompanied by a significant elevation in the number of 50-kHz USVs, relative to administration of vehicle. Under the same experimental conditions, acute amphetamine (2 mg/kg i.p.) robustly elevated the number of 50-kHz USVs emitted by rats, although it did not affect the maximum peak frequency and bandwidth of USVs. Thus, both qualitative and quantitative differences in the effects exerted by caffeine and amphetamine on 50-kHz USVs were observed. Taken together, these findings further clarify the features of caffeine-mediated psychopharmacological effects, and may help to elucidate the differences between the central effects of caffeine and those elicited by other psychostimulants. PMID- 19545476 TI - NK1 receptor antagonism and the neural processing of emotional information in healthy volunteers. AB - The neuropeptide substance P and its receptor NK1 have been implicated in emotion, anxiety and stress in preclinical studies. However, the role of NK1 receptors in human brain function is less clear and there have been inconsistent reports of the value of NK1 receptor antagonists in the treatment of clinical depression. The present study therefore aimed to investigate effects of NK1 antagonism on the neural processing of emotional information in healthy volunteers. Twenty-four participants were randomized to receive a single dose of aprepitant (125 mg) or placebo. Approximately 4 h later, neural responses during facial expression processing and an emotional counting Stroop word task were assessed using fMRI. Mood and subjective experience were also measured using self report scales. As expected a single dose of aprepitant did not affect mood and subjective state in the healthy volunteers. However, NK1 antagonism increased responses specifically during the presentation of happy facial expressions in both the rostral anterior cingulate and the right amygdala. In the emotional counting Stroop task the aprepitant group had increased activation in both the medial orbitofrontal cortex and the precuneus cortex to positive vs. neutral words. These results suggest consistent effects of NK1 antagonism on neural responses to positive affective information in two different paradigms. Such findings confirm animal studies which support a role for NK1 receptors in emotion. Such an approach may be useful in understanding the effects of novel drug treatments prior to full-scale clinical trials. PMID- 19545477 TI - Cocaine-mediated synaptic potentiation is absent in VTA neurons from mGlu5 deficient mice. AB - Drugs of abuse have the ability to instantiate plastic adaptations within the central nervous system, and this property may relate to the development and persistence of addiction. In this context, a single exposure to cocaine in rodents may induce synaptic plasticity by increasing the AMPA/NMDA receptor excitatory post-synaptic current (EPSC) amplitude ratio in dopaminergic cells of the ventral tegmental area (VTA). Here, we examine the role of the metabotropic glutamate 5 (mGlu5) receptor in this regard using a genetic mouse model. The control AMPA/NMDA EPSC ratio is reduced in mGlu5-deficient mice compared to wild types. Moreover, cocaine-induced enhancement of this EPSC ratio is also absent in mutant mice, which suggests that mGlu5 receptors are required for single-dose cocaine-induced plasticity onto VTA cells. While the temporal profile of hyperactivity to acute cocaine is altered in mGlu5-deficient mice; these mice still develop and express sensitized psychomotor responses to cocaine. These data suggest that the mGlu5 receptor is required for cocaine-induced plasticity in VTA dopaminergic cells. In contrast, the mGlu5 receptor may not be essential for psychostimulant behavioural sensitization; although it probably impacts other aspects drug addiction, such as motivation to self-administer. PMID- 19545475 TI - Subchronic SSRI administration reduces insula response during affective anticipation in healthy volunteers. AB - The anterior cingulate cortex (ACC) and insula are important neural substrates for the integration of cognitive, emotional, and physiological information, as well as the coordination of responses to anticipated stimuli. Increased neural activation within these structures has been observed in individuals with anxiety and depressive disorders. Selective serotonin reuptake inhibitors (SSRIs) are among the most effective and frequently prescribed anxiolytic agents, yet it is not known whether ACC or insula underlie the effects of these drugs. We examined whether subchronic administration of a SSRI to healthy volunteers attenuates activation in ACC or insula during anticipation, an important emotional process underlying anxiety. Support for this hypothesis would help to understand where and by what process SSRIs may exert beneficial effects as anxiolytics and would provide further mechanistic evidence for functional magnetic resonance imaging (fMRI) as a biomarker for the development of anxiolytics. Fifteen volunteers participated in a double-blind, placebo-controlled, randomized cross-over study. Participants completed a pleasant and aversive picture-cued anticipation task during fMRI after taking either escitalopram (10 mg) or placebo for 21 d. We found that escitalopram significantly decreased activation in bilateral posterior and middle insula during the anticipation condition irrespective of stimulus valence and in medial prefrontal and ACC during anticipation of aversive vs. pleasant images. Reduced insular and ACC activation in healthy controls during anticipation may be integral to the therapeutic efficacy of SSRIs and may provide a mechanistic approach for the use of pharmaco-fMRI in the identification of novel pharmacotherapeutic agents in patient populations. PMID- 19545479 TI - Anger treatment in chemically-dependent inpatients: evaluation of phase effects and gender. AB - BACKGROUND: There is a growing quest for anger management techniques especially in underserved populations. Patients with a substance abuse history often have untreated anger problems. AIMS: To test a new comprehensive program for prevention, intervention, and remediation of anger in chemically-dependent patients. A secondary aim was to explore any anger differences between males versus females. METHOD: Twenty-six participants (13 male, 13 female) completed three phases of treatment plus follow-up. Dependent measures were six subscales of the (STAXI) questionnaire and self-monitored frequency, duration, and intensity of anger. RESULTS: A significant multivariate effect of phase of study accounted for 42% of the variance in STAXI scores. Univariate F-tests confirmed significant changes on all STAXI subscales. Most of these were between pre and post phases of the study, the effect sizes = +0.8 for state anger and +0.99 for trait anger. For self-monitored variables, significant reductions emerged between treatment phases, the average pre-post effect size = +1.02. Gender did not affect STAXI scores although females had more self-monitored anger, particularly anger episodes. CONCLUSIONS: Findings suggest cumulative efficacy of the anger treatment program. That trait anger declined more than state anger may indicate characterological change in addition to situational change; anger frequency and duration declined more than intensity in keeping with other reports that intensity peaks suddenly and is less modifiable. That males and females were generally similar in anger is worth noting in relation to other studies. Finally, participant attrition is discussed as a problem and a possible index of treatment outcome. PMID- 19545478 TI - Neuroendocrine and neurochemical impact of aggressive social interactions in submissive and dominant mice: implications for stress-related disorders. AB - Social conflicts may engender stress-related behavioural and physiological disturbances in the victims of aggression. In addition, stress-like neurochemical changes and ensuing depressive and anxiety symptoms might also be evident in the perpetrators of aggressive acts. The present investigation assessed basal levels of circulating corticosterone and of brain serotonin (5-HT) and norepinephrine (NE) in pre-identified submissive and dominant mice. In addition, brain neurochemical changes were determined following a single or three 15-min aggressive episodes both in submissive mice and in those that dominated the aggressive interplay. Three minutes after single and repeated confrontations, plasma corticosterone levels and 5-HT utilization within the prefrontal cortex (PFC) and hippocampus were increased to a comparable extent in submissive and dominant animals. Interestingly, however, NE utilization within the PFC and hippocampus was augmented to a greater level in submissive mice. These results suggest that 5-HT neuronal functioning was generally responsive to aggressive events, irrespective of social rank, whereas NE neuronal activity within the PFC and hippocampus was more sensitive to the submissive/dominance attributes of the social situation. It is possible that NE and 5-HT variations associated with an aggressive experience contribute to depressive- and anxiety-like manifestations typically observed after such psychosocial stressors, particularly in submissive mice. However, given that 5-HT changes occur irrespective of social rank, these data suggest that a toll is taken on both submissive and dominant mice, leaving them vulnerable to stress-related pathology. PMID- 19545480 TI - Who attends and who benefits from CBT "self-confidence" workshops run in routine practice? A pilot study. AB - BACKGROUND: This study examined characteristics of members of the public who self referred and the effectiveness of psycho-educational CBT self-confidence workshops when run in routine practice. METHOD: Repeated measures were employed at pre- and post-workshop stages. RESULTS: Of the 56 members of the general public who self-referred to the workshops, 70% were above the clinical cut-offs for Global Distress (CORE OM) and 86% were above the clinical cut offs for depression symptomatology (CES-D). Follow up data (n = 31) showed significant reduction in self-reported distress and depression at 4-week follow-up. A further analysis showed that those whose scores were above the clinical threshold at initial presentation benefited most but those with scores below the threshold did not seem to benefit. CONCLUSIONS: This study demonstrates that psycho-educational CBT workshops attract those with high levels of distress and depression, and have potential as a cost effective means of disseminating psychological interventions. PMID- 19545481 TI - Mindfulness groups for distressing voices and paranoia: a replication and randomized feasibility trial. AB - BACKGROUND: The clinical literature cautions against use of meditation by people with psychosis. There is, however, evidence for acceptance-based therapy reducing relapse, and some evidence for clinical benefits of mindfulness groups for people with distressing psychosis, though no data on whether participants became more mindful. AIMS: To assess feasibility of randomized evaluation of group mindfulness therapy for psychosis, to replicate clinical gains observed in one small uncontrolled study, and to assess for changes in mindfulness. METHOD: Twenty-two participants with current distressing psychotic experiences were allocated at random between group-based mindfulness training and a waiting list for this therapy. Mindfulness training comprised twice-weekly sessions for 5 weeks, plus home practice (meditation CDs were supplied), followed by 5 weeks of home practice. RESULTS: There were no significant differences between intervention and waiting-list participants. Secondary analyses combining both groups and comparing scores before and after mindfulness training revealed significant improvement in clinical functioning (p = .013) and mindfulness of distressing thoughts and images (p = .037). CONCLUSIONS: Findings on feasibility are encouraging and secondary analyses replicated earlier clinical benefits and showed improved mindfulness of thoughts and images, but not voices. PMID- 19545482 TI - A pilot study of telephone cognitive-behavioural therapy for obsessive-compulsive disorder in young people. AB - BACKGROUND: Cognitive-behaviour therapy (CBT) is the recommended psychological treatment for obsessive compulsive disorder (OCD) in young people. Access to CBT may be limited by a number of factors, including lack of trained therapists, and geographic or financial factors preventing access to a specialized service. Telephone delivery of CBT represents one way of overcoming some of these accessibility issues. This pilot study describes outcomes for a telephone-based cognitive-behavioural treatment for obsessive-compulsive disorder (OCD) in young people. METHOD: Ten participants, aged 13 to 17 years, and their parents received up to 16 sessions of telephone CBT (TCBT). Measures of OCD symptoms were obtained using multiple informants and a repeated measures design. Assessments were conducted at pre-treatment, post-treatment, and at 6- and 12-month follow-up. RESULTS: Improvements were found for OCD symptoms across all informants. Family satisfaction with treatment over the telephone was high. CONCLUSIONS: The findings suggest that TCBT is a clinically effective, feasible and acceptable means of service delivery that offers the potential to make CBT a more accessible treatment for young people. TCBT requires further evaluation in randomized, controlled trials to compare effectiveness with face-to-face CBT, which currently represents the usual care model. PMID- 19545483 TI - Cognitive processes during acute psychosis: the role of heightened responsibility and catastrophic misinterpretations. AB - BACKGROUND: This study investigated the role of cognitive mechanisms underlying obsessive compulsive and panic disorders in psychosis, and in particular, their possible contributions to acute psychosis. METHOD: A total of 90 participants were recruited comprising three equal-size groups, including two clinical groups (acute and stable) and one non-clinical matched control group. Symptom severity and distress was assessed using the PSYRATS, and questionnaire measures of anxiety and obsessive beliefs were administered to all participants. RESULTS: Individuals with a diagnosis of psychosis reported significantly higher levels of obsessional beliefs and anxiety sensitivity than the non-clinical group. Furthermore, acutely psychotic patients reported a significantly higher sense of responsibility and catastrophic misinterpretation than the stable psychiatric controls, and than samples of OCD and GAD patients. CONCLUSIONS: Results suggest that these anxiety processes are particularly important during acute psychotic episodes, beyond the reported comorbidity. The theoretical and clinical implications of these findings, the limitations of the methodology employed, and suggestions for future research are discussed. PMID- 19545484 TI - Investigating the relationship between item exposure and test overlap: item sharing and item pooling. AB - To date, exposure control procedures that are designed to control item exposure and test overlap simultaneously are based on the assumption of item sharing between pairs of examinees. However, examinees may obtain test information from more than one examinee in practice. This larger scope of information sharing needs to be taken into account in refining exposure control procedures. To control item exposure and test overlap among a group of examinees larger than two, the relationship between the two indices needs to be identified first. The purpose of this paper is to analytically derive the relationships between item exposure rate and each of the two forms of test overlap, item sharing and item pooling, for fixed-length computerized adaptive tests. Item sharing is defined as the number of common items shared by all examinees in a group, while item pooling is the number of overlapping items that an examinee has with a group of examinees. The accuracy of the derived relationships was verified using numerical examples. The relationships derived will lay the foundation for future development of procedures to simultaneously control item exposure and item sharing or item pooling among a group of examinees larger than two. PMID- 19545485 TI - Children's aggressive behaviour and teacher-child conflict in kindergarten: is teacher perceived control over child behaviour a mediating variable? AB - BACKGROUND: Research repeatedly showed young children's aggressive behaviour to predict relationship difficulties with the teacher. AIMS: To examine a possible mediating variable in this process and in the stability of relationship difficulties across the school year, namely teacher perceived control over child behaviour. SAMPLE: The sample consisted of 139 Belgian kindergartners and their teachers. METHOD: Data were collected throughout kindergarten at three measurement occasions: children's aggressive behaviour was measured by means of a peer nomination procedure during the first trimester, teacher perceived control over child behaviour was assessed by means of a teacher questionnaire during the second trimester, and teacher-child conflict was measured by means of a teacher questionnaire during the first and third trimesters. Correlations among all study variables were calculated and different models were estimated and compared by means of structural equation modelling. RESULTS: Teacher perceived control completely mediated the relationship between aggressive behaviour and teacher child conflict (after controlling for the concurrent association between aggressive behaviour and conflict, and the stability of conflict). In addition, teacher perceived control accounted for part of the stability in conflict across the school year. CONCLUSIONS: Teacher perceived control over child behaviour has been found to act as a mediating mechanism between child aggressive behaviour and teacher-child conflict at the beginning of kindergarten and teacher-child conflict at the end of the year. PMID- 19545486 TI - Expression pattern of dipeptidyl peptidase IV activity and/or structure homologues in cancer. AB - Proline at the second position of the N-terminus of biologically active peptides involved in cell growth regulation is an evolutionarily conserved motif protecting them against cleavage by non-specific proteases. Just a small number of proline-specific hydrolases including dipeptidyl peptidase IV (DPP-IV) and related molecules is capable of cleaving such post-prolyl bond. DPP-IV, originally described on the basis of its enzymatic activity, is a ubiquitous, multifunctional homodimeric plasma membrane glycoprotein of type II. Subsequently, several other molecules related to DPP-IV by their enzymatic activity and/or sequence were discovered and classified as "dipeptidyl peptidase IV activity and/or structure homologues" (DASH). Along with canonical DPP-IV this group comprises DPP-IVbeta, DPP-II, DPP6, DPP8, DPP9, DPP10 and fibroblast activation protein alpha (FAP-alpha). Recent observations of deregulated expression of several DASH molecules in multiple human cancers led to the assumptions of their pathogenetic relevance in cancerogenesis. Here we review recent information about selected DASH molecules in human malignancies. PMID- 19545487 TI - Characterization of human gene expression changes after olive oil ingestion: an exploratory approach. AB - Olive oil consumption is protective against risk factors for cardiovascular and cancer diseases. A nutrigenomic approach was performed to assess whether changes in gene expression could occur in human peripheral blood mononuclear cells after oli ve oil ingestion at postprandial state. Six healthy male volunteers ingested, at fasting state, 50 ml of olive oil. Prior to intervention a 1-week washout period with a controlled diet and sunflower oil as the only source of fat was followed. During the 3 days before and on the intervention day, a very low phenolic compound diet was followed. At baseline (0 h) and at post-ingestion (6 h), total RNA was isolated and gene expression (29,082 genes) was evaluated by microarray. From microarray data, nutrient-gene interactions were observed in genes related to metabolism, cellular processes, cancer, and atherosclerosis (e.g. USP48 by 2.16; OGT by 1.68-fold change) and associated processes such as inflammation (e.g. AKAP13 by 2.30; IL-10 by 1.66-fold change) and DNA damage (e.g. DCLRE1C by 1.47; POLK by 1.44- fold change). When results obtained by microarray were verified by qRT-PCR in nine genes, full concordance was achieved only in the case of up-regulated genes. Changes were observed at a real-life dose of olive oil, as it is daily consumed in some Mediterranean areas. Our results support the hypothesis that postprandial protective changes related to olive oil consumption could be mediated through gene expression changes. PMID- 19545488 TI - Leukaemia inhibitory factor (LIF) gene mutations in women diagnosed with unexplained infertility and endometriosis have a negative impact on the IVF outcome. A pilot study. AB - The frequency of functionally relevant mutations of the leukaemia inhibitory factor (LIF) gene in infertile women is significantly enhanced in comparison with fertile controls. The objective of this retrospective cohort study was to evaluate the impact of LIF gene mutations on the outcome of the treatment in women with various causes of infertility. Fifteen infertile women with the G to A transition at position 3400 leading to the valine to methionine exchange at codon 64 were analysed. Group A was made up of women with diagnoses that are frequently accompanied by changes in humoral as well as cell-mediated immunity - idiopathic infertility and endometriosis (N = 7). Group B consisted of patients with polycystic ovary syndrome (PCOS), andrological factor, tubal factor and hyperprolactinaemia (N = 8). The control group comprised 136 infertile women with no LIF gene mutation diagnosed with idiopathic infertility and endometriosis (N = 37) (group C) and patients with PCOS, tubal and andrological factor (N = 99) (group D). Seven of the mutation-positive patients were successfully treated by in vitro fertilization (IVF), but nobody in this group was diagnosed with idiopathic infertility and only one with endometriosis, which means that there is a statistically significant difference in the pregnancy rates between groups A and B (P = 0.01, Fisher's 2 by 2 exact test) but no statistically significant difference when comparing patients with the LIF gene mutation (group A+B) to no LIF gene mutation (group C+D). The results suggest that in mutation-positive women the idiopathic infertility and endometriosis have a negative impact on the outcome of IVF treatment. PMID- 19545489 TI - Gene expression responses in larvae of the fleshfly Sarcophaga bullata after immune stimulation. AB - Insect larvae develop in decaying organic matter and their defence against various microorganisms must therefore be highly efficient. In the present study, we explored the transcriptional kinetics and induction levels of eight genes in Sarcophaga bullata larvae after infection or aseptic injury. Using real-time PCR, we studied the time-dependent immune response of larvae of the fleshfly S. bullata. We compared the mRNA levels of eight selected genes in induced and non induced larvae. The third-instar larvae of S. bullata were induced by injecting a bacterial suspension of Escherichia coli, Staphylococcus aureus or Pseudomonas aeruginosa, or by simple aseptic injury with an entomological pin. We used intact larvae as a control for basal mRNA expression. Total RNA was isolated from the whole body, fat body and haemocytes. We determined the mRNA levels of genes encoding sapecin, transferrin, prophenoloxidase 1 and 2, storage-binding protein, cathe psin L, sarcocystatin, and 26/29 kDa protease. We found that there was massive up-regulation of genes encoding the fleshfly peptide sapecin, as well as the protein transferrin. We also detected down-regulation of, or no change in, the expression of genes that encode prophenoloxidase 1 and 2, storage-binding protein, cathepsin L, sarcocystatin, and 26/29 kDa protease. PMID- 19545490 TI - Differences between molecular mechanisms involved in the regulation of haptoglobin gene expression during the acute phase response and dietary restriction. AB - Haptoglobin is a glycoprotein involved in the acute phase response. Previously we reported that haptoglobin gene expression was up-regulated during dietary restriction in young female rats. The present study aimed at determining whether chronic dietary restriction affects haptoglobin blood levels through changing levels and/or activities of IL-6-related transcription factors STAT and C/EBP in the liver as is the case during the acute phase response. To this end, we compared a female Wistar rat model of 50% 6-week-long dietary restriction with the standard laboratory model for the acute phase response induced by turpentine administration. During the turpentine-induced acute phase response, the transitory 5.4-fold increase of rat haptoglobin expression was accompanied by a prominent rise of serum IL-6 concentration and the increased binding of STAT3 and 35kD C/EBPbeta/LAP transcription factors to the haptoglobin gene hormone responsive element. Results obtained after immunoblotting and DNA affinity chromatography (using hormone-responsive element) suggest that the stable 1.7 fold increase of serum haptoglobin level during dietary restriction was the result of increased amounts and activities of constitutive transcription factors C/EBPalpha and STAT5b, and to a smaller extent of STAT3. When dietary restriction rats were administered turpentine, a 8.7-fold increase in haptoglobin expression was followed by a considerable increase in the amount and hormone-responsive element binding activity of STAT3 but not 35kD C/EBPbeta/LAP. We concluded that haptoglobin gene up-regulation during chronic dietary restriction was regulated by different mechanisms than during the acute phase response, and that it depended on the amount(s) and activit(ies) of transcription factor(s) that characterize low-grade inflammatory conditions. PMID- 19545492 TI - Prevention of catheter-related infection: evidence-based medicine. PMID- 19545491 TI - Apolipoprotein E Arg136 --> Cys in individuals with premature myocardial infarction. AB - Coronary artery disease is a serious health problem worldwide caused by interactions between genetic and environmental risk factors. One of the candidate genes is the gene for apolipoprotein E. We present a case report of two young smoking and obese carriers (man 45 years and woman 32 years old) of the apolipoprotein E (p.Arg136Cys) mutation, but with no severe dyslipidaemias detected among 1,671 survivors (1,483 men, 188 women, aged 21-75 years) of acute coronary syndrome screened for genetic and traditional cardiovascular risk factors. Between acute coronary syndrome survivors, the mutation has not yet been described. Even though this mutation raises suspicion to be a risk factor for cardiovascular disease (based on previous publications), its frequency was very low and similar to the control population (12 detected carriers of the mutation within the 9,386 screened individuals). Therefore, whether this rare mutation is causal for the development of myocardial infarction needs to be further evaluated. PMID- 19545493 TI - Vaccination for the prevention of pneumococcal meningitis. PMID- 19545494 TI - Cardiac complications of infective endocarditis. AB - Cardiac complications caused by infective endocarditis (IE) are varied and frequently life-threatening. This article focuses on new data related to several complications, and summarizes the indications, timing, and type of valve surgery recommended in the management of IE. Several recent studies using propensity score techniques have resulted in disparate conclusions and underscore the need for randomized prospective studies to better address whether and when surgery should be performed in patients with IE. Mitral valve repair is an exciting new development in surgical methodology and probably will have increased application over the next two decades. Excellent reviews related to periannular abscesses, fistulae, acute coronary syndrome, and pericarditis have been published recently and are also summarized. PMID- 19545495 TI - Cardiac assist device infections. AB - Cardiac assist devices have emerged as an increasingly important option for circulatory support in patients with advanced congestive heart failure. Infection has been the leading cause of death with use of ventricular assist devices for the past 25 years, and continues to be a major cause of morbidity and mortality in this population. Infections may involve the internal (blood-containing) components, or occur on the external device surfaces. The pump pocket and the percutaneous driveline are particularly vulnerable. In this article we review the pathogenesis, epidemiology, clinical course, and approaches for treatment and prevention of cardiac assist device infections. PMID- 19545497 TI - Bacterial endophthalmitis. AB - Endophthalmitis refers to bacterial or fungal infection of the vitreous and/or aqueous humors of the eye. Bacterial endophthalmitis occurs most commonly after eye surgery or penetrating ocular trauma (exogenous endophthalmitis), but may also occur from hematogenous seeding during bacteremia (endogenous endophthalmitis). The presentation is usually acute, with eye pain and decreased vision. In exogenous endophthalmitis, infection is confined to the eye. There is no fever and minimal, if any, peripheral leukocytosis. Treatment includes direct injection of antibiotics into the vitreous, and vitrectomy in more severe cases. Systemic antibiotics are indicated in endogenous endophthalmitis; their role in exogenous endophthalmitis is controversial. Visual outcome depends on the virulence of the bacterial pathogen and the speed with which treatment is given. Acute bacterial endophthalmitis is a medical emergency, because delay in treatment may result in vision loss. PMID- 19545496 TI - Diagnosis and management of mycotic aneurysms. AB - We reviewed all papers most recently reported in the literature related to infected arterial aneurysms (IAAs) affecting the aorta and vascular beds other than the aorta. In this article, we report on vascular beds other than the aorta. As is the case for aortic IAAs, infected non-aortic aneurysms are rarely encountered. The majority of recent studies are limited to case reports and small series of unusual infected aneurysms. A comprehensive review of this entity was performed based on the available literature from January through December 2008 in all languages. Available reports were analyzed with respect to demographic features, type of presentation, methods of diagnosis and therapy, follow-up, and outcome (morbidity and mortality). PMID- 19545498 TI - Coccidioidal meningitis: update on epidemiology, clinical features, diagnosis, and management. AB - Coccidioidomycosis is a fungal infection endemic to geographically restricted areas of the southwestern United States, and its incidence is increasing within endemic areas. Coccidioidal meningitis, an infrequent but very severe form of coccidioidomycosis, is associated with significant mortality and morbidity. The diagnosis depends on the demonstration of coccidioidal antibody or positive culture, histopathology, or cytology in the presence of typical abnormalities of the cerebrospinal fluid. Central nervous system complications of this infection may include hydrocephalus, vasculitis, infarctions, or parenchymal abscesses. Many patients with coccidioidal meningitis can achieve remission of infection following treatment with triazole antifungal agents. However, first-line therapy with high-dose oral fluconazole is associated with a high incidence of relapse after drug discontinuation. This article summarizes the pathogenesis, epidemiology, clinical manifestations, diagnostic studies, and treatment of coccidioidal meningitis, and highlights new information regarding this devastating infection. PMID- 19545499 TI - Update on rabies diagnosis and treatment. AB - The diagnosis of rabies depends on recognizing the clinical picture, because a history of an animal bite may be present or absent. Laboratory tests, particularly when based on detection of rabies virus antigen or RNA in tissues or fluids, often confirm the diagnosis. Negative laboratory tests do not exclude the diagnosis unless they are performed on brain tissues. Preventive therapy, including wound cleansing and active and passive immunization after a recognized exposure, is well established and highly efficacious. No established therapy exists that is effective for patients with rabies encephalomyelitis. The Milwaukee protocol involves induction of therapeutic coma; however, there is no clear rationale for a neuroprotective role of this therapy, many reports exist of its failures, and its use should be abandoned. Basic research is needed on the mechanisms of rabies pathogenesis. This may allow the development of new therapeutic approaches for this ancient disease. PMID- 19545501 TI - Antiretroviral therapy for treatment-naive patients: a review of recent literature and the updated guidelines. AB - The improved efficacy and safety of antiretroviral agents have fundamentally changed the face of HIV infection. Advances in antiretroviral therapy have dramatically improved the prognosis of HIV infection, which is now on a par with other chronic illnesses. This article addresses the optimal timing of antiretroviral therapy initiation and the optimal drug choices for initial regimens, with a focus on recent data and published guidelines issued by the US Department of Health and Human Services and the International AIDS Society-USA. PMID- 19545500 TI - Bacterial meningitis vaccines: not just for kids. AB - Bacterial meningitis remains a feared diagnosis that may lead to significant mortality and morbidity in both the developed and developing world; case fatality rates range from 10% to 50% among all age groups. Several vaccines are available (including the Haemophilus influenzae type B polysaccharide conjugate vaccine) that have proven effective in reducing the incidence of bacterial meningitis. Although a 23-valent pneumococcal polysaccharide vaccine has been available for some time, use of the 7-valent pneumococcal conjugate vaccine (PCV) has demonstrated a dramatic reduction in the incidence of invasive pneumococcal disease; despite the increase in the disease caused by nonvaccine-serotype strains, the success of the 7-valent PCV is noteworthy. A quadrivalent conjugate meningococcal vaccine has been available in the United States since 2005; although its true efficacy remains to be determined, there have been dramatic results with declining incidence in certain high-risk populations and in endemic areas. PMID- 19545502 TI - Prevention counseling for HIV-infected persons: what every clinician needs to know. AB - The absence of adequate and regular prevention counseling during routine clinical encounters translates into missed opportunities for HIV prevention. HIV care providers have considerably more contact with patients than clinicians in other disciplines. These contacts should be translated into opportunities to provide HIV prevention messages to patients and should be a priority for all clinicians caring for HIV-positive patients. Coincidental preventive care for HIV-positive patients is inherently unproductive because of the absence of reinforcing messages. In a recent meta-analysis, HIV-positive individuals who underwent counseling and testing services reduced high-risk behaviors by about 68%. Prevention counseling should focus on positive reinforcement, harm reduction, education, and support. We strongly recommend regular, brief, targeted prevention counseling as a part of every clinical encounter. PMID- 19545503 TI - Newer antiretroviral agents and how to use them. AB - Several relatively new antiretroviral drugs have been approved or are under investigation. These include nucleoside and nonnucleoside reverse transcriptase inhibitors, protease and integrase strand transfer inhibitors, CCR5 antagonists, and an entirely new class of maturation inhibitors. Although most of these drugs were developed for patients with drug-resistant HIV-1, some have demonstrated a potential role for those starting treatment for the first time. This review incorporates data from the most recent treatment guidelines, peer-reviewed publications, and recent presentations at research meetings about some of these relatively new and emerging antiretroviral agents and offers strategies for incorporating them into clinical practice. PMID- 19545504 TI - Interspecies extrapolation in environmental exposure standard setting: A Bayesian synthesis approach. AB - Currently the extrapolation of evidence from studies of non-human species to the setting of environmental exposure standards for humans includes the imposition of a variety of uncertainty factors reflecting unknown aspects of the procedure, including the relevance of evidence from one species to impacts in another. This paper develops and explores more flexible modelling of aspects of this extrapolation, using models proposed by DuMouchel [DuMouchel, W.H., Harris, J.E., 1983. Bayes methods for combining the results of cancer studies in humans and other species (with comment). J. Am. Statist. Assoc. 78, 293-308.] The approaches are based on Bayesian meta-analysis methods involving explicit modelling of relevance in the prior distributions, estimated using Markov chain Monte Carlo (MCMC) methods. The methods are applied to evidence relating chlorinated by products exposure to adverse reproductive health effects. The relative merits of various approaches are discussed, and developments and next steps are outlined. PMID- 19545505 TI - Veterinary diclofenac threatens Africa's endangered vulture species. AB - Veterinary diclofenac has been responsible for the devastation of three species of Gyps vulture on the Indian subcontinent, and it is now regarded as one of the worst environmental contaminants in the recent past. While measures have been taken to control the manufacture of veterinary diclofenac in South Asia, the promotion of diclofenac on the African continent poses a risk to vultures in this region. In Southern Africa, the species of greatest conservation concern is the Cape Griffon Vulture (Gyps coprotheres), as only 2900 breeding pairs remain in the wild. The objective of this study was to test if this species is toxicologically sensitive to diclofenac. In a single dose-toxicity study, two adult Cape Griffon Vultures with severe injuries, that were considered to have a very poor prognostic outcome, were dosed intravenously with diclofenac at 0.8mg/kg. The changes in the clinical pathology were compared to the normal reference range established for 24 healthy Cape Griffon Vultures. Both birds died within 48h of dosing. The clinical signs, clinical pathology, gross pathology and histopathological finding were typical for diclofenac toxicity. It would appear that the sensitivity of the Cape Griffon is similar to that of their Asian counterparts and the African White-backed Vulture (Gyps africanus). Diclofenac is almost certainly toxic to all Gyps vultures species and strong efforts must be taken to ensure that veterinary diclofenac products are not licensed or introduced to the African continent. PMID- 19545507 TI - Transparency throughout the production chain--a way to reduce pollution from the manufacturing of pharmaceuticals? AB - Recent findings have shown that wastewater from bulk drug production can be a source of very high environmental concentrations of drugs in certain locations. The release of active ingredients is often not specifically regulated, and thus rapid initiatives from the industries themselves are warranted. Possible ways to stimulate action include changes in local and international regulations, including the implementation of appropriate environmental standards within existing industry guidelines as well as demands from prescribers and consumers of medicines. The lack of readily available information regarding the origin of drugs and the environmental impact of the production, however, prevents consumers from making informed decisions. Here, we investigated the origin of active pharmaceutical ingredients (APIs) in 242 selected products on the Swedish market. By comparing registers from Sweden and India we found that the APIs in 71 products (31%) originated from Indian manufacturers sending their waste to a treatment plant where unprecedented amount of environmental pollution with broad spectrum antibiotics and other drugs recently has been documented. We propose that increased transparency throughout the production chain would be one of several important steps to reducing pollution from the manufacturing of drugs. PMID- 19545509 TI - Proteomics tools and resources for investigating protein allergens in oilseeds. AB - Oilseeds are important renewable sources of natural products including protein and oil which are produced during the maturation (or seed filling) phase of embryo development. My lab employed high-resolution, two-dimensional gel electrophoresis (2-DE) and mass spectrometry to profile and identify over 500 proteins expressed during seed filling in various oilseeds including soybean, canola, castor, and Arabidopsis. The principal objective of these studies was to develop predictive models for carbon assimilation for comparison among the four oilseeds. Other uses for these large proteomic datasets have come to light including characterization of the diversity and expression of known and yet-to-be discovered protein allergens as they accumulate during seed development. Legume oilseeds such as soybean and peanut present a human and animal health concern for a small percentage of the population that are allergic to one or more of the seed proteins. Information about the expression and diversity of 2-DE spots that map to individual genes or gene families of allergens can prove useful for breeding- or biotechnology-based approaches aimed at silencing allergen expression. We have begun releasing these proteomics datasets for public access on the Oilseed Proteomics web portal, www.oilseedproteomics.missouri.edu. I will present the status of these projects and the website with specific emphasis on soybean. PMID- 19545511 TI - Reconstruction of historical exposures in the US nickel alloy industry and the implications for carcinogenic hazard and risk assessments. AB - Recently, various regulatory authorities have been reexamining the potential carcinogenic hazards and risks associated with exposures to nickel and certain nickel compounds. In making their assessments, the authorities have focused on occupational cohorts at facilities where nickel-containing sulfidic ores were processed and where increased lung and nasal cancer risks were found in specific groups of workers. Little attention, however, has been paid to the vast number of workers in nickel-using industries, where no excess respiratory cancer risks have been observed. In this paper, the historical exposures of one such group of workers engaged in the production of nickel alloys are reconstructed, and the implications for cancer risk assessments are analyzed. The results indicate that nickel alloy workers were exposed to insoluble oxidic and metallic nickel species at levels comparable to those found in certain nickel processing cohorts; yet they experienced no increase in respiratory cancer risks. This suggests that extrapolating risks from certain primary nickel producers to other nickel industry sectors may not be appropriate. PMID- 19545510 TI - Development of PBPK model of molinate and molinate sulfoxide in rats and humans. AB - Molinate has been widely used as a pre-emergent herbicide in the rice fields of California's Central Valley. In rat studies, the metabolite molinate sulfoxide is suspected of causing testicular toxicity after exposure to molinate. The sulfoxide is generated in the liver and can circulate in the blood, eventually reaching the testis. Man qualitatively produces the same molinate metabolites as the rat. To extrapolate the reproductive risk to man, the present study outlines the development of a preliminary PBPK (physiologically-based pharmacokinetic) model, validation in the rat and extrapolation to man. The preliminary seven compartment PBPK model for molinate was constructed for the adult, male Sprague Dawley rat that employed both flow-limited (blood, kidney, liver, rapid-perfused tissues and slowly perfused tissues) and diffusion-limited (fat) rate equations. The systemic circulation connects the various compartments. The simulations predict the molinate blood concentrations of the rat blood and testes compartment favorably with the profiles obtained from 10 and 100mg/kg po or 1.5 and 15mg/kg iv doses. Human physiological parameters were substituted into the oral dosed model and the simulations closely predicted the molinate blood concentration obtained from 5.06mg oral dose. A sensitivity analysis determined for an oral dose that peak blood molinate concentrations were most responsive to the blood flows to kidney and fat compartments while testicular molinate sulfoxide concentrations depended on molinate sulfoxide partition coefficients for the testes compartment and the K(m) for glutathione conjugation of molinate sulfoxide in the liver compartment. PMID- 19545513 TI - Risk assessment of ciprofloxacin, flavomycin, olaquindox and colistin sulfate based on microbiological impact on human gut biota. AB - Trace levels of veterinary antibiotics that reside in livestock products may disturb the balance of human intestinal microbiota and impair the colonized barrier function, which is critical to protect against the invasion or overgrowth of exogenous pathogens. We investigated the colonization barrier disruption effect of ciprofloxacin, flavomycin, olaquindox and colistin sulfate by the minimum inhibitory concentration (MIC) assay in pure culture of human gut bacteria and evaluated the no-observed-effect-concentration (NOEC) and acceptable daily intake (ADI) based on the microbiological impact. MICs of the antibiotics were tested for total 100 isolates composed of 10 isolates from each of 10 predominant genera of human faeces that were freshly collected from healthy women at 1x10(5) and 1x10(9) colony-forming units (CFU)/ml. MIC assay was also conducted with 10 ATCC standard bacteria species of human fecal microbiota for the comparison with freshly isolated human fecal mirobiota. The most susceptible bacteria were Escherichia coli for ciprofloxacin and colistin sulfate, Fusobacterium spp. for flavomycin and Eubacterium spp. for olaquindox. MIC values were lower at 1x10(5) than at 1x10(9)CFU/ml. The susceptibility of feacal microbiota freshly isolated from healthy women tended to be similar with those of ATCC standard bactera. NOEC (microg/ml) and ADI (microg/kg BW/day) were evaluated as 0.008 and 0.15 for ciprofloxacin, 0.25 and 1 for flavomycin, 0.125 and 3 for olaquindox and 1.0 and 7 for colistin sulfate, respectively. PMID- 19545514 TI - Safety evaluation of arachidonic acid rich Mortierella alpina biomass in albino rats--a subchronic study. AB - Safety evaluation of arachidonic acid rich Mortierella alpina biomass was carried out in Wistar rats by acute and subchronic oral toxicity studies. A preliminary acute toxicity study revealed that the biomass was safe at acute doses and that the LD50 exceeded 5000mg/kg BW, the highest dose used in the study. In subchronic study, rats were fed diet containing 0, 2500, 5000, 10,000, 20,000 and 30,000mg/kg, M. alpina biomass for a period of 13 weeks. Results indicated that biomass fortification had a positive influence on growth with no overt toxic effects on the survival, food consumption and body weight gain throughout the treatment interlude. The statistically significant changes in relative organ weights, serum biochemical and hematological indices in M. alpina fed groups' viz., higher relative weights of spleen, liver, brain and ovary in females, reduced hemoglobin concentration in males, elevated WBC counts at highest dose, reduction in serum triglycerides and increased alkaline phosphatase activity were not concomitant with pertinent histopathological changes and hence toxicologically inconsequential. No microscopic or macroscopic lesions attributable to the treatment were manifested in the experimental groups. The results of the present study strongly advocate the safety of M. alpina biomass in rats at levels used in the study. PMID- 19545515 TI - Cryptosporidium in birds, fish and amphibians. AB - Whilst considerable information is available for avian cryptosporidiosis, scant information is available for Cryptosporidium infections in fish and amphibians. The present review details recent studies in avian cryptosporidiosis and our current knowledge of piscine and amphibian infections. PMID- 19545516 TI - Minireview: clinical cryptosporidiosis. AB - Cryptosporidium has emerged as an important cause of diarrhoeal illness worldwide, especially amongst young children and patients with immune deficiencies. Usually presenting as a gastro-enteritis-like syndrome, disease ranges in seriousness from mild to severe and signs and symptoms depend on the site of infection, nutritional and immune status of the host, and parasite related factors. Sources and routes of transmission are multiple, involving both zoonotic and anthroponotic spread, and facilitated by the resistance of the parasite to many commonly used disinfectants. Prevention and control measures are important for the protection of vulnerable groups since treatment options are limited. This review covers the life cycle, pathogenesis, clinical presentations, diagnosis, prevention and management of cryptosporidiosis in humans. PMID- 19545520 TI - Identification and expression analysis of the Steinernema carpocapsae elastase like serine protease gene during the parasitic stage. AB - A cDNA encoding elastase was isolated from Steinernema carpocapsae by suppression subtractive hybridization and rapid amplification of 5' cDNA ends. The predicted protein contained a 19-aa signal peptide, a 44-aa N-terminal propeptide, and a 264-aa mature protein with a predicted molecular mass of 28,949 Da and a theoretical pI of 8.88. BLAST analysis showed 27-35% amino acid sequence identity to serine proteases from insects, mammals, fish and other organisms. The Sc-ela gene contains three exons and two introns with at least two copies in the S. carpocapsae genome. Expression analysis indicated that the Sc-ela gene was upregulated during the initial parasitic stage. Sequence comparison and evolutionary marker analysis revealed that Sc-ELA was a member of the elastase serine protease family with potential degradative, developmental and fibrinolytic activities. Homology modeling showed that Sc-ELA adopts a two beta-barrel fold typical of trypsin-like serine proteases, and phylogenetic analysis indicates that Sc-ELA branched off early during elastase evolution. PMID- 19545519 TI - Development of species-specific PCR and PCR-restriction fragment length polymorphism assays for L.infantum/L.donovani discrimination. AB - Discrimination of Leishmania infantum and L. donovani, the members of the L. (L.) donovani complex, is important for diagnosis and epidemiological studies of visceral leishmaniasis (VL). We have developed two molecular tools including a restriction fragment length polymorphisms of amplified DNA (PCR-RFLP) and a PCR that are capable to discriminate L. donovani from L. infantum. Typing of the complex was performed by a simple PCR of cysteine protease B (cpb) gene followed by digestion with DraIII. The enzyme cuts the 741-bp amplicon of L. donovani into 400 and 341 bp fragments whereas the 702 bp of L. infantum remains intact. The designed PCR species-specific primer pair is specific for L. donovani and is capable of amplifying a 317 bp of 3' end of cpb gene of L. donovani whereas it does not generate an amplicon for L. infantum. The species-specific primers and the restriction enzyme were designed based on a 39 bp insertion/deletion (indel) in the middle of the cpb gene. Both assays could differentiate correctly the two species and are reliable and high-throughput alternatives for molecular diagnosis and epidemiological studies of VL in various foci. PMID- 19545521 TI - Toxoplasma gondii: sensitive and rapid detection of infection by loop-mediated isothermal amplification (LAMP) method. AB - Loop-mediated isothermal amplification (LAMP) method amplifies DNA with high specificity, sensitivity and rapidity. In this study, we used a conserved sequence in the 200- to 300-fold repetitive 529 bp gene of Toxoplasma gondii to design primers for LAMP test. Detection limit of T. gondii LAMP assay with the primers is 1 pg/microL of T. gondii DNA, which was evaluated using 10-fold serially diluted DNA of cultured parasites. Furthermore, LAMP and conventional PCR methods were applied for amplification of the T. gondii DNA extracted from the lymph nodes taken from pigs which were suspected to be Toxoplasma infection. As a result, 76.9% (70/91) and 85.7% (78/91) of the samples were positive on PCR and LAMP analyzes, respectively. Therefore, the LAMP has a potential to be applied as an alternative molecular diagnostic tool for detection of T. gondii infection from veterinary samples. This is the first study, which applies the LAMP method to diagnose Toxoplasma from veterinary samples. PMID- 19545523 TI - Toxoplasma gondii: proteomic analysis of antigenicity of soluble tachyzoite antigen. AB - The obligate intracellular parasite Toxoplasma gondii is an important pathogen of humans and animals. The tachyzoite of T. gondii is the main life-cycle stage that is responsible for toxoplasmosis. Study of the antigenicity of soluble tachyzoite antigen (STAg) is important for discovery of protective antigens which will aid in the detection and prevention of toxoplasmosis. At present, no complete proteome map of T. gondii STAg is established, although a large-scale whole proteomic analysis of tachyzoites is underway. In this study, 1227 protein spots of T. gondii soluble tachyzoite antigen (STAg) were fractionated by 2-dimensional electrophoresis (2-DE) at pH range 3-10. By mass spectrometry (MS) analysis, among the separated 1227 protein spots, 426 were identified by searching the Swissport and NCBI nr databases. Two hundred and thirty of these identified spots (230/426, 54%) were demonstrated to be T. gondii protein by MS. Of the 21 Toxoplasma protein spots identified by Western blot with rabbit anti-T. gondii serum, 16 had immunoregulatory functions and five had immune defense functions. Due to multiple spots for a single protein, these 16 spots represented 11 proteins: a putative protein disulfide isomerase (PDI), heat shock protein 60 (Hsp60), a pyruvate kinase (PK), a putative glutamate dehydrogenase (GDH), a coronin, a heat shock protein 70 (Hsp70), a protein kinase C receptor 1 (RACK1), a malate dehydrogenase (MDH), a major surface antigen 1 (SAG1), an uridine phosphorylase (UPase) and a peroxiredoxin (Prx). Among the identified 11 proteins, except that the antigenicity and immunogenicity of the SAG1 has been reported and antigenicity of Hsp70 has been disputed, the remaining antigenic proteins were first identified in this study. In conclusion, we obtained nine novel types of immunogenic proteins that might be potential candidates of vaccine development for toxoplasmosis, which we will confirm in later studies. PMID- 19545522 TI - Trypanosoma cruzi: antiproliferative effect of indole phytoalexins on intracellular amastigotes in vitro. AB - American trypanosomiasis (Chagas disease) continues to be a significant public health problem, and the therapeutic potential of current antichagasic agents (nifurtimox and benznidazole) is rather limited. Here we report on the antitrypanosomal effect of 1-methoxyspirobrassinol and other indole phytoalexins- secondary metabolites produced by Cruciferous plants. These compounds, that previously demonstrated antimicrobial and anticancer properties, displayed significant antiproliferative effects on intracellular amastigotes of Trypanosoma cruzi and may be prospective candidates for antichagasic drug design and development. PMID- 19545524 TI - Toxocara canis: larval migration dynamics, detection of antibody reactivity to larval excretory-secretory antigens and clinical findings during experimental infection of gerbils (Meriones unguiculatus). AB - In this study, Mongolian gerbils were used to analyse features of Toxocara infection that included larval migration, humoral immune responses to Toxocara canis excretory-secretory antigens (TES) and aspects of host physiology. At day 10 post-infection (p.i.) most larvae were in the intestine and the lungs while later the total number of larvae was higher in the carcass tissue; the number of larvae per gram of tissue was lower elsewhere other than in the brain. Infected animals showed several neurological abnormalities, an early increase in leukocyte and neutrophil levels, two peaks of peripheral eosinophilia (5 and 40 d.p.i.) and high antibody levels against TES in the circulation and in the vitreous humor. A sequential recognition of eight T.canis larval antigens with MW from 24 to 200 kDa was detected by Western blot. The results obtained in this study further support the use of gerbils as an experimental model for systemic, ocular and cerebral toxocariasis. PMID- 19545525 TI - Real-time PCR assays for monitoring benzimidazole resistance-associated mutations in Ancylostoma caninum. AB - Frequent and broad application of anthelmintic drugs for treatment of intestinal parasite infection has led to drug resistance that often renders whole populations of livestock unresponsive to treatment. Therefore, it is important to detect mutations associated with drug resistance before it becomes clinically manifest. To monitor developing drug resistance against benzimidazoles (BZ), we developed real-time PCR assays and applied them to analyse the beta-tubulin isotype-1 gene of the hookworm Ancylostoma caninum, an important parasite of dogs. Previously, we developed PCR assays to monitor codon positions 167 and 200. Here, we describe an assay which is able to detect resistance alleles in codon 198. These real-time PCR assays were subsequently applied to screen hookworm specimens recovered from dogs in Georgia. No elevated levels of polymorphisms at the investigated loci were found, suggesting that selection for resistance in the tested samples did not occur. PMID- 19545528 TI - Giardia lamblia: immunogenicity and intracellular distribution of GHSP-115, a member of the Giardia head-stalk family of proteins. AB - Giardia lamblia, a protozoan causing diarrheal outbreaks, is one of the main pathogens monitored in developed countries. Immunoscreening of G. lamblia expression library using the monoclonal antibodies (mAb) against G. lamblia, identified a subset of antigenic proteins in this protozoan, which are proteins belonging to GHSP (Giardia head-stalk protein), GHSP115, GHSP138, and GHSP180. In order to map the epitope region of GHSP115, the corresponding open reading frame was dissected into three parts and expressed as recombinant proteins with histidine tags. Western blot analysis of these recombinant proteins with mAbs reacting with GHSP115 indicated that one-third of the C-terminus of GHSP115 showed immunoreactivity with the mAb. Intracellular location of GHSP115 was examined both in trophozoites and encysting cells of G. lamblia by an immunofluorescence assay, indicating that location of GHSP115 varies during encystation. These results suggest that GHSP115 is an abundant and antigenic protein, which is differentially localized during life cycle of G. lamblia. PMID- 19545527 TI - Malaria antigen-mediated enhancement of interleukin-21 responses of peripheral blood mononuclear cells in African adults. AB - We recently showed that IL-21 is associated with high level of anti-EBA-175 IgG1 and IgG3. Here we have investigated the ability of two malarial antigens, Glutamate-rich protein and merozoite surface protein 3 to induce IL-21 production from PBMCs from malaria-exposed and non-exposed donors. We found that malaria exposed donors produced significantly more IL-21 compared to non-exposed donors. These data suggest that IL-21 could be involved in the acquisition of immunity to malaria. PMID- 19545529 TI - Trypanosoma cruzi: participation of cholesterol and placental alkaline phosphatase in the host cell invasion. AB - One of the most important parasitic endemic diseases in Latin America is Chagas disease, with almost 20 million people being infected and 60 million others at risk of infection. In the cell infection by Trypanosoma cruzi, the first step is contact with the host cell by receptors and ligands on the membrane. It is known that lipids play an important role in the interaction process between pathogens and host cells with lipid rafts being highly specialized regions of the plasma membrane that are enriched in cholesterol and sphingolipids. We explored whether the treatment with methyl-beta-cyclodextrin alone or by adding Mevinolin, an inhibitor of cholesterol synthesis could deplete membrane cholesterol of the HEp2 cell and if this treatment could affect the trypomastigote infection into the host cell. These treatments led to a leakage of cholesterol, and concomitantly, PLAP enzyme and unidentified proteins resulting in a decrease of the invasion process. However, the GGTP enzyme was not liberated from the host cell membranes. PMID- 19545532 TI - Cryptosporidiosis and giardiasis in dogs and cats: veterinary and public health importance. AB - Dogs and cats are the only domestic animals that still routinely reside in the same domicile as their owners around the world, and hence the interest in their role as reservoirs of potentially zoonotic agents. In the case of cryptosporidiosis and giardiasis, current data suggests that dogs and cats do not routinely share their infections with healthy people. Dogs are hosts of Cryptosporidiumcanis and Giardiaduodenalis Assemblages C and D. Cats are hosts to Cryptosporidiumfelis and G. duodenalis Assemblage F. Dogs and cats (and other animals) are sometimes infected with sub-Assemblage AI, an Assemblage also found in people, but people usually have sub-Assemblage AII. Unfortunately, severely immunocompromised individuals and malnourished children can be made ill by infections with C. canis and C. felis. People should practice good sanitation and hygiene to minimize environmental contamination and contact with the infectious (oo)cysts that may be shed by their pets. PMID- 19545530 TI - Developmental expression analysis and immunolocalization of a biogenic amine receptor in Schistosoma mansoni. AB - A Schistosoma mansoni G-protein coupled receptor (SmGPCR) was previously cloned and shown to be activated by the biogenic amine, histamine. Here we report a first investigation of the receptor's subunit organization, tissue distribution and expression levels in different stages of the parasite. A polyclonal antibody was produced in rabbits against the recombinant third intracellular loop (il3) of SmGPCR. Western blot studies of the native receptor and recombinant protein expressed in HEK293 cells showed that SmGPCR exists both as a monomer (65 kDa) and an apparent dimer of approximately 130 kDa These species were verified by immunoprecipitation of SmGPCR from S. mansoni extracts, using antibody that was covalently attached to agarose beads. Further investigation determined that the SmGPCR dimer was resistant to treatment with various detergents, 4 M urea and 0.1 M DTT but could be made to dissociate at acidic pH, suggesting the dimer is non covalent in nature. Confocal immunofluorescence studies revealed significant SmGPCR immunoreactivity in sporocysts, schistosomula and adult worms but not miracidia. SmGPCR was found to be most widely expressed in the schistosomula, particularly the tegument, the subtegumental musculature and the acetabulum. In the adult stage we detected SmGPCR immunofluorescence mainly in the tubercles of male worms and, to a lesser extent, the body wall musculature. Localization in sporocysts was mainly confined to the tegument and cells within parenchymal matrices. A real-time quantitative reverse-transcription PCR analysis revealed that SmGPCR is upregulated at the mRNA level in the parasitic stages compared to the free-living miracidium and cercariae, and it is particularly elevated during early sporocyst and schistosomula development. The results identify SmGPCR as an important parasite receptor with potential functions in muscle and the tegument of S. mansoni. PMID- 19545534 TI - Expression of peptidylarginine deiminase from Porphyromonas gingivalis in Escherichia coli: enzyme purification and characterization. AB - Porphyromonas gingivalis peptidylarginine deiminase (PAD) catalyzes the deimination of peptidylarginine residues of various peptides to produce peptidylcitrulline and ammonia. P. gingivalis is associated with adult-onset periodontitis and cardiovascular disease, and its proliferation depends on secretion of PAD. We have expressed two recombinant forms of the P. gingivalis PAD in Escherichia coli, a truncated form with a 43-amino acid N-terminal deletion and the full-length form of PAD as predicted from the DNA sequence. Both forms contain a poly-His tag and Xpress epitope at the N-terminus to aid in detection and purification. The activities and stabilities of these two forms have been evaluated. PAD is cold sensitive; it aggregates within 30 min at 4 degrees C, and optimal storage conditions are at 25 degrees C in the presence of a reducing agent. PAD is not a metalloenzyme and does not need a cofactor for catalysis or stability. Multiple l-arginine analogs, various arginine-containing peptides, and free l-arginine were used to evaluate substrate specificity and determine kinetic parameters. PMID- 19545535 TI - Redox properties of the oxygen-detoxifying flavodiiron protein from the human parasite Giardia intestinalis. AB - Flavodiiron proteins (FDPs) are enzymes identified in prokaryotes and a few pathogenic protozoa, which protect microorganisms by reducing O(2) to H(2)O and/or NO to N(2)O. Unlike most prokaryotic FDPs, the protozoan enzymes from the human pathogens Giardia intestinalis and Trichomonas vaginalis are selective towards O(2). UV/vis and EPR spectroscopy showed that, differently from the NO consuming bacterial FDPs, the Giardia FDP contains an FMN with reduction potentials for the formation of the single and the two-electron reduced forms very close to each other (E(1)=-66+/-15mV and E(2)=-83+/-15mV), a condition favoring destabilization of the semiquinone radical. Giardia FDP contains also a non-heme diiron site with significantly up-shifted reduction potentials (E(1)=+163+/-20mV and E(2)=+2+/-20mV). These properties are common to the Trichomonas hydrogenosomal FDP, and likely reflect yet undetermined subtle structural differences in the protozoan FDPs, accounting for their marked O(2) specificity. PMID- 19545536 TI - Photoaffinity labeling the agonist binding domain of alpha4beta4 and alpha4beta2 neuronal nicotinic acetylcholine receptors with [(125)I]epibatidine and 5[(125)I]A-85380. AB - The development of nicotinic acetylcholine receptor (nAChR) agonists, particularly those that discriminate between neuronal nAChR subtypes, holds promise as potential therapeutic agents for many neurological diseases and disorders. To this end, we photoaffinity labeled human alpha4beta2 and rat alpha4beta4 nAChRs affinity-purified from stably transfected HEK-293 cells, with the agonists [(125)I]epibatidine and 5[(125)I]A-85380. Our results show that both agonists photoincorporated into the beta4 subunit with little or no labeling of the beta2 and alpha4 subunits respectively. [(125)I]epibatidine labeling in the beta4 subunit was mapped to two overlapping proteolytic fragments that begin at beta4V102 and contain Loop E (beta4I109-P120) of the agonist binding site. We were unable to identify labeled amino acid(s) in Loop E by protein sequencing, but we were able to demonstrate that beta4Q117 in Loop E is the principal site of [(125)I]epibatidine labeling. This was accomplished by substituting residues in the beta2 subunit with the beta4 homologs and finding [(125)I]epibatidine labeling in beta4 and beta2F119Q subunits with little, if any, labeling in alpha4, beta2, or beta2S113R subunits. Finally, functional studies established that the beta2F119/beta4Q117 position is an important determinant of the receptor subtype-selectivity of the agonist 5I-A-85380, affecting both binding affinity and channel activation. PMID- 19545537 TI - Solid-liquid interface method (SLIM): a new crystallization method for proteins. AB - Despite impressive advances in theories, methods and technologies, crystallization still remains a serious bottleneck in structural determination of macromolecules. Here we present a novel solid-liquid interface method (SLIM) for protein crystallization, based on the pre-adding and drying of a crystallization reagent, and thereafter the dispensing of a protein solution to the dried media to initiate crystallization from the solid-liquid interface. Not only quick and easy to perform, the method also allows for a less concentrated protein solution for setting up crystallization trials. PMID- 19545538 TI - Neuromyelitis optica: Passive transfer to rats by human immunoglobulin. AB - Recurrent attacks of optic neuritis and myelitis are the hallmarks of both neuromyelitis optica (NMO) and multiple sclerosis (MS). NMO immunoglobulin G (NMO IgG), which recognizes astrocytic aquaporin-4 (AQP4) water channels, is a specific serum autoantibody that distinguishes NMO from MS. The pathogenic role of the anti-AQP4 antibody (AQP4-Ab, NMO-IgG) in NMO has been speculated based on several studies in vitro. The aim of this study was to demonstrate the pathogenicity of AQP4-Ab in vivo. We obtained IgG from patients who underwent therapeutic plasmapheresis, and developed an animal model by passive transfer of IgG to rats. The active lesions of the rats exhibited pathological characteristics strikingly similar to those of NMO, marked by astrocytic loss and perivascular deposition of immunoglobulin and complements. These findings provide the first evidence of the pathogenicity of AQP4-Ab in vivo and support the therapeutic efficacy of eliminating the antibodies by plasmapheresis. PMID- 19545539 TI - Development of a novel pretargeting system with bifunctional nucleic acid molecules. AB - This study was aimed at exploring a novel pretargeting system based upon bifunctional nucleic acid molecules that are comprised of a nucleic acid aptamer and a nucleic acid tail. The properties of bifunctional molecules were investigated by both theoretical prediction and experimental determination. Different from the algorithm-based structure prediction, the experimental data showed that some nucleic acid tails could significantly decrease the binding capability of the aptamer. It was also found that the effectiveness of bifunctional molecules in labeling cells was dependent on the hybridization length. Based on these understandings, one bifunctional molecule was selected to study pretargeting. The results demonstrated that the bifunctional molecule could not only bind to target cells, but also hybridize with its complementary oligonucleotide on the cell surface. Thus, bifunctional nucleic acid molecules hold great potential for pretargeting applications. PMID- 19545540 TI - Paradoxical effect of L-arginine: acceleration of endothelial cell senescence. AB - We have recently shown that inhibition of nitric oxide (NO) synthesis by asymmetrical dimethylarginine (ADMA) accelerated endothelial cell (EC) senescence which was prevented by coincubation with L-arginine; however the effect of long term treatment of l-arginine alone on senescence of ECs have not been investigated. Human ECs were cultured in medium containing different concentrations of L-arginine until senescence. L-Arginine paradoxically accelerated senescence indicated by inhibiting telomerase activity. Moreover, L arginine decreased NO metabolites, increased peroxynitrite, and 8-iso prostaglandin F(2alpha) formation. In old cells, the mRNA expression of human amino acid transporter (hCAT)2B, the activity and protein expression of arginase II were upregulated indicated by enhanced urea, L-ornithine, and L-arginine consumption. Inhibition of arginase activity, or transfection with arginase II siRNA prevented L-arginine-accelerated senescence. The most possible explanation for the paradoxical acceleration of senescence by L-arginine so far may be the translational and posttranslational activation of arginase II. PMID- 19545541 TI - Focal adhesion kinase (FAK) activates and stabilizes IGF-1 receptor. AB - Recent studies have shown a direct association between IGF-1R and FAK, two important mediators of cell growth, survival and migration. However, the mechanism by which FAK affects IGF-1R function remains unknown. This study investigates the potential role of FAK in mediating activation and stability of IGF-1R. Autophosphorylation and phosphorylation capacities of wild type and mutant IGF-1R were studied. Surprisingly, we found that the mutant IGF-1R lacking the three core tyrosine residues in the activation-loop can be phosphorylated although it is unable to undergo autophosphorylation, suggesting that another kinase possesses the ability to phosphorylate IGF-1R. By using wild type MEFs and FAK-/- MEFs we could demonstrate that FAK mediates activation-loop independent phosphorylation, as well as Akt and ERK activation. Furthermore, the stability of IGF-1R was decreased upon FAK siRNA or inactivation. Taken together, our data suggest a role for FAK in phosphorylation, signaling and stability of the IGF-1R. PMID- 19545542 TI - Inhibition of canonical Wnt signaling promotes gliogenesis in P0-NSCs. AB - Wnt signaling plays an essential role in the development of mammalian central nervous system. We investigated the impact of activation/inhibition of the Wnt signaling pathway on neuronal/glial differentiation in neurospheres derived from neonatal mouse forebrains. For short term alterations, neurospheres were stimulated with recombinant Wnt-3a, Wnt-5a and the Wnt inhibitor Dickkopf-1 (Dkk1). Furthermore, neurospheres were transduced with retroviral vectors encoding Wnt-3a, Wnt-7a and their inhibitors Dkk1 and soluble Frizzled related protein-5 (sFRP5). Long-term activation of Wnt pathway by Wnt-7a or by treatment with GSK3 inhibitors promoted a moderate increase of the neuronal differentiation and blocked gliogenesis. In contrast, Wnt pathway inhibition in neurospheres, induced by retroviral overexpression of either Dkk1 or sFRP5, robustly increased the gliogenesis at the expense of neurogenesis. In summary, our data demonstrate that activation or inhibition of Wnt/beta-catenin signaling in neurospheres regulates neuronal and glial differentiation, respectively. Thus, our results suggest that Wnt signaling may also contribute to regulate these processes in the neonatal brain. PMID- 19545543 TI - Rho-associated kinase (ROCK) inhibition reverses low cell activity on hydrophobic surfaces. AB - Hydrophobic polymers do not offer an adequate scaffold surface for cells to attach, migrate, proliferate, and differentiate. Thus, hydrophobic scaffolds for tissue engineering have traditionally been physicochemically modified to enhance cellular activity. However, modifying the surface by chemical or physical treatment requires supplementary engineering procedures. In the present study, regulation of a cell signal transduction pathway reversed the low cellular activity on a hydrophobic surface without surface modification. Inhibition of Rho associated kinase (ROCK) by Y-27632 markedly enhanced adhesion, migration, and proliferation of osteoblastic cells cultured on a hydrophobic polystyrene surface. ROCK inhibition regulated cell-cycle-related molecules on the hydrophobic surface. This inhibition also decreased expression of the inhibitors of cyclin-dependent kinases such as p21(cip1) and p27(kip1) and increased expression of cyclin A and D. These results indicate that defective cellular activity on the hydrophobic surface can be reversed by the control of a cell signal transduction pathway without physicochemical surface modification. PMID- 19545544 TI - ATX-3, CDC-48 and UBXN-5: a new trimolecular complex in Caenorhabditis elegans. AB - Ataxin-3 is the protein involved in Machado-Joseph disease, a neurodegenerative disorder caused by a polyglutamine expansion. Ataxin-3 binds ubiquitylated proteins and acts as a deubiquitylating enzyme in vitro. It was previously proposed that ataxin-3, along with the VCP/p97 protein, escorts ubiquitylated substrates for proteasomal degradation, although other players of this escort complex were not identified yet. In this work, we show that the Caenorhabditis elegans ataxin-3 protein (ATX-3) interacts with both VCP/p97 worm homologs, CDC 48.1 and CDC-48.2 and we map the interaction domains. We describe a motility defect in both ATX-3 and CDC-48.1 mutants and, in addition, we identify a new protein interactor, UBXN-5, potentially an adaptor of the CDC-48-ATX-3 escort complex. CDC-48 binds to both ATX-3 and UBXN-5 in a non-competitive manner, suggesting the formation of a trimolecular complex. Both CDC-48 and ATX-3, but not UBXN-5, were able to bind K-48 polyubiquitin chains, the standard signal for proteasomal degradation. Additionally, we describe several common interactors of ATX-3 and UBXN-5, some of which can be in vivo targets of this complex. PMID- 19545545 TI - A DNA aptamer recognizes the Asp f 1 allergen of Aspergillus fumigatus. AB - Allergies are caused by the binding of IgE antibodies onto specific sites on allergens. However, in the assessment of exposure to airborne allergens, current techniques such as whole spore counts fail to account for the presence of these allergenic epitopes that trigger allergic reactions. The objective of the research is to develop a DNA aptamer for the Asp f 1 allergen of the pathogenic fungus Aspergillus fumigatus, using an IgE-binding epitope of the allergen as the target for aptamer selection. Through in vitro SELEX, an aptamer has been produced that binds with nanomolar affinity to the Asp f 1 IgE-epitope. The aptamer is also able to recognize the native Asp f 1 allergen, and does not bind to allergenic proteins from non-target mold species such as Alternaria alternata. Production of this aptamer provides proof-of-principle that allergen measurement methods can be developed to indicate the potent fraction, or allergenicity, of allergens. PMID- 19545546 TI - Modulation of glucocorticoid receptor nuclear translocation in neurons by immunophilins FKBP51 and FKBP52: implications for major depressive disorder. AB - Mood disorders associated with dysfunction of the hypothalamic-pituitary-adrenal (HPA) axis are common psychiatric conditions. The glucocorticoid receptor (GR) is a steroid-activated nuclear receptor that, upon binding to cortisol, translocates to the nucleus where it targets genes related to neuronal metabolism and plasticity. In patients suffering from major depressive disorder (MDD), hypercortisolemia is a common finding. In the current study we investigated the molecular events associated with the FK506 binding proteins (FKBP) -52 and -51 response to cortisol exposure in neuronal cell cultures and their effect on GR translocation. We noted that FK506 altered nuclear localization of the GR and inhibited expression of GR-responsive genes. Furthermore, siRNA knockdown of FKBP4 gene, coding for the immunophilin FKBP52, inhibited cortisol-activated GR nuclear translocation, while knockdown of FKBP5, coding for immunophilin FKBP51, was associated with increased baseline GR nuclear localization. We propose that immunophilins are modulators of the cortisol-HPA axis response to stress and related chronic brain disorders. PMID- 19545547 TI - Chronic, systemic treatment with a metabotropic glutamate receptor 5 antagonist in 6-hydroxydopamine partially lesioned rats reverses abnormal firing of dopaminergic neurons. AB - Increasing evidence indicates that the excessive glutamate release onto substantia nigra pars compacta (SNpc) dopaminergic neurons may play an important role in the progression of nigral degeneration. We examined the effects of chronic, systemic treatment with 2-methyl-6-(phenylethynyl)-pyridine (MPEP), a selective metabotropic glutamate receptor 5 antagonist, in firing activity of SNpc dopaminergic neurons in 6-hydroxydopamine (6-OHDA) partially lesioned rats. In 6-OHDA-lesioned rats treated with vehicle, injection of 6-OHDA (4 microg) into the medial forebrain bundle produced a partial lesion, 39% loss of tyrosine hydroxylase-immunoreactive (TH-ir) neurons in the SNpc. In partially lesioned rats, the electrophysiological characteristics of SNpc dopaminergic neurons showed that the firing rate of these neurons increased compared with sham operated rats and the firing pattern also changed towards a burstier one. Chronic, systemic treatment of MPEP (3 mg/kg/day, 14 days) attenuated loss of TH ir neurons and normalized the hyperactive firing activity in the SNpc induced by partial unilateral dopamine depletion lesions. In addition, no significant differences were found in the responsiveness of SNpc dopaminergic neurons to intravenous cumulative apomorphine in sham-operated, vehicle-treated and MPEP treated rats, while ED50 values for apomorphine in MPEP-treated rats were decreased as compared with vehicle-treated rats. These data demonstrate that the surviving dopaminergic neurons in the SNpc are hyperactive in an experimental model of moderate Parkinson's disease (PD). In this model, chronic, systemic MPEP treatment has the neuroprotective effect and reverses the abnormal firing activity of dopaminergic neurons, suggesting that MPEP has an important implication for the treatment of PD. PMID- 19545549 TI - Role of preoptic opioid receptors in the body temperature reduction during hypoxia. AB - Evidence indicates that endogenous opioids play a role in body temperature (Tb) regulation in mammals but no data exist about the involvement of the specific opioid receptors, mu, kappa and delta, in the reduction of Tb induced by hypoxia. Thus, we investigated the participation of these opioid receptors in the anteroventral preoptic region (AVPO) in hypoxic decrease of Tb. To this end, Tb of unanesthetized Wistar rats was monitored by temperature data loggers before and after intra-AVPO microinjection of the selective kappa-opioid receptor antagonist nor-binaltorphimine dihydrochloride (nor-BNI; 0.1 and 1.0 microg/100 nL/animal), the selective mu-opioid receptor antagonist D-Phe-Cys-Tyr-D-Trp-Arg Thr-Pen-Thr-NH2 cyclic (CTAP; 0.1 and 1.0 microg/100 nL/animal), and the selective delta-opioid receptor antagonist Naltrindole (0.06 and 0.6 microg/100 nL/animal) or saline (vehicle, 100 nL/animal), during normoxia and hypoxia (7% inspired O2). Under normoxia, no effect of opioid antagonists on Tb was observed. Hypoxia induced Tb to reduce in vehicle group, a response that was inhibited by the microinjection intra-AVPO of nor-BNI. In contrast, CTAP and Naltrindole did not change Tb during hypoxia but caused a longer latency for the return of Tb to the normoxic values just after low O2 exposure. Our results indicate the kappa opioid receptor in the AVPO is important for the reduction of Tb during hypoxia while the mu and delta receptors are involved in the increase of Tb during normoxia post-hypoxia. PMID- 19545548 TI - Opposing actions of neuronal nitric oxide synthase isoforms in formalin-induced pain in mice. AB - The role of central and peripheral neuronal nitric oxide synthase (nNOS) splice variants in the development of inflammatory hyperalgesia was investigated using the formalin test. Supraspinal administration of the NOS inhibitor NOArg lowered both the first and second phase of the formalin response. An oligodeoxynucleotide targeting four nNOS isoforms given supraspinally also reduced the formalin response of both phases. Supraspinal antisense mapping suggested that this effect results from the nNOS-1 splice variant, implying that nNOS-1 is important in mediating formalin pain. At the spinal level, antisense mapping suggested a role of both the nNOS-1 and the nNOS-beta variants in producing formalin pain. Conversely, an antisense selective against nNOS-2 had an opposing effect against the first phase, increasing its intensity. This result, which was similar to prior studies examining opioid actions, implies that endogenous nNOS-2 activity acted to minimize pain perception. Locally in the foot, arginine, the precursor for NO, increased the phase II response at low doses while higher doses reduced the response. This complex biphasic response suggested opposing NOS actions. Local antisense mapping again showed that nNOS-1 is involved in producing phase II of the formalin response while nNOS-2 had an opposite effect similar to that seen spinally. Finally, downregulation of nNOS-1 by antisense prevented tolerance to morphine in both the tail-flick and the formalin test. Together, these observations illustrate the complexity of nNOS in pain perception and the existence of opposing nNOS systems likely due to splice variants of nNOS. PMID- 19545550 TI - Increased serum HMGB1 is related to the severity of coronary artery stenosis. AB - BACKGROUND: High mobility group box 1 (HMGB1) protein has been identified as a novel pro-inflammatory cytokine in coronary artery disease (CAD). We investigated the relationship between serum HMGB1 level and the severity of coronary artery stenosis. METHODS: Serum HMGB1 concentration in patients was measured by ELISA. All patients underwent coronary angiography and were assigned into groups according to the quartile of HMGB1 level. RESULTS: HMGB1 level in USAP group was higher than that in control and SAP group. HMGB1 level in SAP group was higher than that in control group (p<0.05). Gensini scores in the highest quartile group (group IV), group III and group II were all significantly higher than that in the lowest quartile group (group I). There was significant correlation between angiographic Gensini score and serum level of HMGB1 (r=0.710, p<0.05). However, in subgroup analysis, we found that serum HMGB1 level was only correlated with angiographic Gensini score in SAP patients (r=0.786, p<0.05), but not USAP patients. CONCLUSIONS: Serum HMGB1 level was markedly increased with the severity of coronary artery stenosis in patients with SAP and USAP, especially in SAP patients, which suggested that increased serum HMGB1 may involve in the pathogenesis of atherosclerotic CAD. PMID- 19545551 TI - TNFalpha promoter polymorphism is a risk factor for susceptibility in hepatocellular carcinoma in Korean population. AB - BACKGROUND: The underlying genetic factors for the development and progression of hepatocellular carcinoma (HCC) are largely unknown. TNFalpha is a well characterized inflammatory mediator and is implicated in the development of HCC. We investigated TNFalpha polymorphisms for association with HCC. METHODS: The study population consisted of 227 HCC patients and 365 age and sex matched Korean controls. TNFalpha polymorphisms (G-238A, C-857T, and C-863A) were genotyped using pyrosequencing analysis. TNFalpha levels in patients with HCC were determined by enzyme linked immunosorbent assay (ELISA). Logistic regression analysis was used to determine the association with HCC and haplotype was calculated using EH program. RESULTS: Of three TNFalpha polymorphisms investigated in our study, C-863A did not correlate with HCC. However, both G 238A and C-857T were found to be significantly associated with HCC. TNFalpha 238A allele was more frequent in HCC patients than in control [P=0.012; odds ratio (OR), 1.89; 95% confidence interval (CI), 1.14-3.13]. TNFalpha -857T was significantly associated with HCC patients (P=0.001; OR, 1.63; 95% CI, 1.21 2.19). Haplotype analysis revealed that the GTC haplotype (G-238A, C-857T, C 863A) was a risk marker for HCC (P=0.0021). Serum TNFalpha level was significantly increased in HCC patients with CT+TT genotype for TNFalpha -857 (P=0.018). CONCLUSION: Our data imply that TNFalpha G-238A and C-857T, not C 863A, polymorphisms may confer different susceptibilities to the development of HCC with TNFalpha -238A and -857T alleles playing as risk factors. PMID- 19545553 TI - Improved MS/MS analysis of succinylacetone extracted from dried blood spots when combined with amino acids and acylcarnitine butyl esters. AB - BACKGROUND: The utilization of succinylacetone (SUAC) as the primary metabolic marker for tyrosinemia Type I is now well known, thus new methods have been developed to analyze SUAC as a first tier test in newborn screening. One approach is to prepare a SUAC hydrazine derivative from the dried blood spots (DBS) previously utilized in the extraction of acylcarnitine (AC) and amino acids (AA). The final derivatized products of SUAC, AA and AC are combined in a single tandem mass spectrometric (MS/MS) analysis. However, butyl esterification techniques may result in contamination of underivatized acylcarnitines by as much as 20%. We have developed a simple wash step to improve the combined analysis of SUAC, AA and AC in DBS by MS/MS. METHODS: AA and AC were extracted with methanol containing labeled internal standard from 3.2mm punches taken from the DBS specimen. The previously extracted blood spot that remains after removal of the methanol extraction solvent was used in the preparation of SUAC with and without additional washing of the blood spot. The butyl ester eluates of AA and AC, and SUAC hydrazine derivatives were recombined and measured by MS/MS. RESULTS: Three additional methanol wash steps of the remaining DBS punches prior to SUAC derivatization reduced the presence of underivatized acylcarnitines, resulting in a 4-fold reduction of underivatized palmitoylcarnitine. Palmitoylcarnitine butyl ester is detected at m/z 456 while the underivatized species is detected at m/z 400, which is also the mass of dodecanoylcarnitine butyl ester. The linearity of the SUAC assay was unchanged by the additional wash steps. For butyl esterification methods, the preferred analytic procedure, the presence of AC can compromise the results of a newborn screen for the actual concentrations of acylcarnitines. It is essential to remove any underivatized acylcarnitines prior to SUAC analysis. CONCLUSION: The additional methanol wash steps did not alter SUAC assay results but did remove underivatized acylcarnitines which could result in the incorrect quantification of acylcarnitines. PMID- 19545552 TI - Binding of the monomeric form of C-reactive protein to enzymatically-modified low density lipoprotein: effects of phosphoethanolamine. AB - BACKGROUND: The 5 subunits of native pentameric C-reactive protein (CRP) are dissociated to generate the monomeric form of CRP (mCRP) in some in vitro conditions, both physiological and non-physiological, and also in vivo. Many bioactivities of mCRP generated by urea-treatment of CRP and of mCRP generated by mutating the primary structure of CRP have been reported. The bioactivities of mCRP generated by spontaneous dissociation of CRP are largely unexplored. METHODS: We purified mCRP generated by spontaneous dissociation of CRP and investigated the binding of mCRP to enzymatically-modified low-density lipoprotein (E-LDL). RESULTS: mCRP was approximately 60 times more potent than CRP in binding to E-LDL. In the presence of the small-molecule compound phosphoethanolamine (PEt), at 37 degrees C, the binding of mCRP to E-LDL was enhanced <2-fold, while the binding of CRP to E-LDL was enhanced >10-fold. In contrast, PEt inhibited the binding of both CRP and mCRP to pneumococcal C polysaccharide, another phosphocholine-containing ligand to which CRP and mCRP were found to bind. We have not investigated yet whether PEt alters the structure of CRP at 37 degrees C. CONCLUSIONS: Combined data suggest that the targeting of CRP with the aim to monomerize CRP in vivo may be an effective approach to capture modified forms of LDL. PMID- 19545554 TI - Response to hydroxyurea in beta thalassemia major and intermedia: experience in western India. AB - BACKGROUND: The clinical and hematological response to hydroxyurea was evaluated in beta thalassemia patients in western India with variable clinical severity and correlated with genetic factors. MATERIALS AND METHODS: Seventy-nine patients-[38 beta thalassemia intermedia-(group I), 41-beta thalassemia major-(group II)] on hydroxyurea therapy were followed-up for 20-24months. RESULTS: Among the frequently transfused patients in group I, 58% became transfusion independent and 16% showed a 50% reduction in transfusions after therapy which correlated with a higher mean fold increase in HbF and gamma mRNA expression levels. Forty-one percent of patients in group I had associated alpha-thalassemia and 72.7% were XmnI (+/+). beta thalassemia chromosomes among the responders of group I (41%) were linked to haplotype (- + + - + + - - +) as against haplotype (+ - - - - - - +) being more common among the non-responders. Response was not linked to the beta thalassemia mutations. Thirty-two percent of group II patients showed a 50% reduction in their transfusion requirements after therapy which also correlated with a higher mean fold increase in HbF and gamma mRNA expression levels. A significant decrease in serum ferritin was seen in both groups. 19% of patients could not tolerate the drug. CONCLUSIONS: In group I, clinical response to hydroxyurea was better in patients with alpha-thalassemia, XmnI (+/+) and a higher mean fold increase in gamma mRNA expression. In group II, only one-third of patients showed a partial response. PMID- 19545555 TI - Comparison of performance of three commercial platforms for warfarin sensitivity genotyping. AB - BACKGROUND: We performed a 3-way comparison on the Osmetech eSensor, AutoGenomics INFINITI, and a real-time PCR method (Paragonx reagents/Stratagene RT-PCR platform) for their FDA-cleared warfarin panels, and additional polymorphisms (CYP2C9*5, *6, and 11 and extended VKORC1 panels) where available. METHODS: One hundred de-identified DNA samples were used in this IRB-approved study. Accuracy was determined by comparison of genotyping results across three platforms. Any discrepancy was resolved by bi-directional sequencing. The CYP4F2 on Osmetech was validated by bi-directional sequencing. RESULTS: Accuracies for CYP2C9*2 and *3 were 100% for all 3 platforms. VKORC1 3673 genotyping accuracies were 100% on eSensor and 97% on Infiniti. CYP2C9*5, *6 and *11 showed 100% concordance between eSensor and Infiniti. VKORC1 6484 and 9041 variants compared between ParagonDx and Infiniti analyzer were 100% (6484) and 99% (9041) concordant. CYP4F2 was 100% concordant with sequencing results. The time required to generate the results from automated DNA extraction-to-result was approximately 8h on Infiniti, and 4h on eSensor and ParagonDx, respectively. CONCLUSIONS: Overall, we observed excellent CYP2C9*2 and *3 genotyping accuracy for all three platforms. For VKORC1 3673 genotyping, eSensor demonstrated a slightly higher accuracy than the Infiniti, and CYP4F2 on Osmetech was 100% accurate. PMID- 19545556 TI - Mean telomere length and risk of incident venous thromboembolism: a prospective, nested case-control approach. AB - BACKGROUND: Recent studies have shown telomere-length shortening as a risk predictor for cardiovascular disease. However, to date, no prospective data are available on its potential involvement in venous thromboembolism (VTE). METHODS: Using leukocyte DNA samples collected at baseline in a prospective cohort of 14,916 initially healthy American men, we examined the relationship of mean telomere repeat copy number to single gene copy number (T/S ratio), using a modified quantitative polymerase chain reaction protocol, amongst 108 White males who subsequently developed a first ever VTE event and amongst an equal number of age- and smoking-matched White males who remained free of vascular events during follow-up (controls). RESULTS: An inverse correlation between T/S ratios and age was observed in our controls (p=0.04). However, the T/S ratios were similar between cases and controls (p=0.31). Furthermore, in a multi-variable adjusted analysis, we found no evidence for an association of the observed T/S ratios with VTE risk (odds ratio=1.20; 95%CI=0.58-2.52; p=0.62). CONCLUSIONS: The present investigation found no evidence for an association of relative telomere length with risk of incident VTE. PMID- 19545557 TI - Plasma oxidized lipoprotein(a) and its immune complexes are present in newborns and children. AB - BACKGROUND: Oxidized Lp(a) [ox-Lp(a)] has been reported to play more potent roles than native Lp(a) in atherosclerosis. We investigated the distribution characteristics of plasma ox-Lp(a) and Lp(a) immune complex [Lp(a)-IC] levels in newborns and children. METHODS: Plasma ox-Lp(a) and Lp(a)-IC levels were measured in 747 children and 30 cord blood by ELISAs. RESULTS: The mean levels of Lp(a), ox-Lp(a) and Lp(a)-IC were much lower in newborns than in children (P<0.001), and increased rapidly to that in children after birth. The distributions of Lp(a), ox Lp(a) and Lp(a)-IC were skewed toward low values in children, no difference of their levels was found in each of the 13year groups. The levels of ox-Lp(a) correlated positively with total and LDL cholesterol, Lp(a) and Lp(a)-IC; Lp(a) IC correlated positively with sex, total and LDL cholesterol, Lp(a) and ox-Lp(a), respectively. Multiple linear regression analysis showed Lp(a) and Lp(a)-IC accounted for 42% of the variation in ox-Lp(a) levels, and ox-Lp(a) accounted for 30% of that in Lp(a)-IC. CONCLUSIONS: The fact that ox-Lp(a) and Lp(a)-IC are present in newborns and children suggests that oxidized lipoproteins play an initiating role in atherosclerotic process. PMID- 19545558 TI - Comparison of the permeability properties and post-thaw motility of ejaculated and epididymal bovine spermatozoa. AB - There are very few experimental reports on the comparative water transport (membrane permeability) characteristics of ejaculated and epididymal mammalian spermatozoa during freezing. In the present study, we report the effects of cooling ejaculated and epididymal bovine sperm from the same males with and without the presence of a cryoprotective agent, glycerol. Water transport data during freezing of ejaculated and epididymal bovine sperm suspensions were obtained at a cooling rate of 20 degrees C/min under two different conditions: (1) in the absence of any cryoprotective agents, CPAs and, (2) in the presence of 0.7 M glycerol. Using values published in the literature, we modeled the spermatozoa as a cylinder of length 39.8 microm and a radius of 0.4 microm with an osmotically inactive cell volume, V(b), of 0.61 V(o), where V(o) is the isotonic cell volume. The subzero water transport response is analyzed to determine the variables governing the rate of water loss during cooling of bovine spermatozoa, i.e. the membrane permeability parameters (reference membrane permeability, L(pg) and activation energy, E(Lp)). The predicted best-fit permeability parameters ranged from, L(pg)=0.021-0.038 microm/min-atm and E(Lp)=27.8-41.1 kcal/mol. The subzero water transport response and consequently the subzero water transport parameters are not significantly different between the ejaculated and epididymal bovine spermatozoa under corresponding cooling conditions. If this observation is found to be more generally valid for other mammalian species as well, then in the future the sperm extracted from the testes of a postmortem male could be optimally cryopreserved using procedures similar to those derived for ejaculated sperm. PMID- 19545559 TI - Gdf6a is required for the initiation of dorsal-ventral retinal patterning and lens development. AB - Dorsal-ventral patterning of the vertebrate retina is essential for accurate topographic mapping of retinal ganglion cell (RGC) axons to visual processing centers. Bone morphogenetic protein (Bmp) growth factors regulate dorsal retinal identity in vertebrate models, but the developmental timing of this signaling and the relative roles of individual Bmps remain unclear. In this study, we investigate the functions of two zebrafish Bmps, Gdf6a and Bmp4, during initiation of dorsal retinal identity, and subsequently during lens differentiation. Knockdown of zebrafish Gdf6a blocks initiation of retinal Smad phosphorylation and dorsal marker expression, while knockdown of Bmp4 produces no discernable retinal phenotype. These data, combined with analyses of embryos ectopically expressing Bmps, demonstrate that Gdf6a is necessary and sufficient for initiation of dorsal retinal identity. We note a profound expansion of ventral retinal identity in gdf6a morphants, demonstrating that dorsal BMP signaling antagonizes ventral marker expression. Finally, we demonstrate a role for Gdf6a in non-neural ocular tissues. Knockdown of Gdf6a leads to defects in lens-specific gene expression, and when combined with Bmp signaling inhibitors, disrupts lens fiber cell differentiation. Taken together, these data indicate that Gdf6a initiates dorsal retinal patterning independent of Bmp4, and regulates lens differentiation. PMID- 19545560 TI - Autoradiographic comparison of in vitro binding characteristics of various tritiated adenosine A2A receptor ligands in rat, mouse and pig brain and first ex vivo results. AB - The adenosine A(2A) receptor in the basal ganglia is involved in the control of movement and plays a role in movement disorders such as Parkinsonism. Developing ligands to evaluate that receptor by noninvasive methods such as positron emission tomography has a high priority. In vitro radioligand binding guides the selection of ligands for in vivo application. This study measured the binding of the adenosine A(2A) receptor antagonist [(3)H]MSX-2 (3-(3-hydroxypropyl)-8-m methoxystyryl)-7-methyl-1-propargylxanthine) to rat, mouse and pig brain by autoradiography. Other studies measured binding to membranes from PC12 pheochromocytoma cells. Those binding parameters were compared to those of the adenosine A(2A) receptor antagonist [(3)H]ZM241385 (4-(2-[7-amino-2-(2 furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino)ethyl)phenol), the adenosine A(2A) receptor agonist [(3)H]CGS21680 (2-[p-(2-carboxyethyl)-phenethylamino]-5'-N ethylcarboxamidoadenosine) and the unselective adenosine receptor agonist [(3)H]NECA (5'N-ethylcarboxamido)adenosine). The potency order (K(d)) in the three species was [(3)H]ZM241385<[(3)H]MSX-2<[(3)H]NECA<[(3)H]CGS21680. The density of [(3)H]MSX-2 binding sites was greater in the striatum than in the cortex. Preliminary ex vivo experiments showed that by 10min after iv injection, [(3)H]MSX-2 and [(3)H]CGS21680 crossed the blood-brain barrier to the extent of almost 1% ID/g brain tissue, but [(3)H]NECA and [(3)H]ZM241385 to only 0.2% ID/g. The prior administration of unlabeled ZM241385 significantly lowered brain uptake of [(3)H]MSX-2. In conclusion, [(3)H]MSX-2 has a high affinity and sufficient selectivity for the adenosine A(2A) receptor. It penetrates the blood-brain barrier. Sensitivity to photoisomerization is a limitation. Further investigations assess its suitability as a ligand for imaging the brain adenosine A(2A) receptor. PMID- 19545561 TI - Possible contribution of beta-glucosidase and caspases in the cytotoxicity of glufosfamide in colon cancer cells. AB - Glycoconjugates represent a recent trend in cancer chemotherapy that adopts the concept of selective prodrug/drug targeting of tumor cells by binding to specific transmembrane glucose transporters. Following preferential uptake of sugar conjugates into cancer cells, they are presumably subject to enzymatic cleavage by specific beta-glycosidases to liberate the free active cytotoxic aglycones that act selectively on cancer cells and spare other noncancerous ones. In this sense, the role of beta-glucosidase and caspases in the bioactivation and cytotoxicity of glufosfamide has been addressed in the current study. The cytotoxicity of glufosfamide has been investigated over 24-96 h in a panel of human colon cancer cells namely, Caco-2, HT29 and T84 using a tetrazole dye; 3 (4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; MTT assay technique. Apoptosis was assessed by fluorometric assay of caspase-3 and caspase-9 activities. Enzymatic cleavage of glufosfamide was accomplished using a host of hydrolytic enzymes and cleavage kinetics was determined using HPLC. Glufosfamide has proven cytotoxic efficacy in a concentration- and time-dependent manner. The sensitivity rank order of tumor cells towards the glycoconjugate was Caco 2>HT29>T84. This sensitivity ranking was well correlated with the enzymatic activity of beta-glucosidase assessed in these cell lines. Initiation and activation of apoptosis were increased in all colon cancer cells following exposure to glufosfamide and were well correlated with the cytotoxicity rank order of the glycoconjugate. Glufosfamide was cleaved by cytosolic and lysosomal beta-glucosidases but not by other hydrolytic enzymes such as cytosolic beta galactosidase, pancreatic lipase or hepatic esterase. In conclusion, the current data could possibly unravel the mechanistic role of beta-glucosidase and apoptotic caspases in the bioactivation and cytotoxicity of glufosfamide within colon cancer cells. PMID- 19545562 TI - Cilostazol reduces proliferation through c-Myc down-regulation in MDCK cells. AB - Cilostazol, a drug commonly used in the treatment of intermittent claudication is a selective phosphodiesterase III inhibitor. It affects cell proliferation, increases cAMP levels, activates the cyclic AMP-dependent protein kinase and inhibits E2F in vascular cells. Polycystic kidney disease, a common genetic disorder, is characterized by increased cell proliferation, basement membrane abnormalities and fluid secretion. An established in vitro model of this disease is the canine Madin-Darby cell line (MDCK). In this communication, we investigated the effects of cilostazol exposure in MDCK cells. A reduced cell proliferation rate with an arrest in the G1 phase of the cell cycle was detected. Accordingly, several transcription factors associated with cell cycle control were affected by cilostazol, particularly c-myc. c-Myc DNA binding as well as its transcriptional activity was severely impaired in cilostazol-treated cells. Pharmacological tools demonstrated that besides the involvement of the cyclic AMP dependent protein kinase, the extracellular signal-regulated kinases I/II participate in the response. These results suggest that cilostazol inhibits cell proliferation through c-myc transcriptional control, also pave the way to our better understanding of molecular transactions triggered by this drug and strengthen its potential use in other malignancies. PMID- 19545563 TI - Fucoidan protects against dopaminergic neuron death in vivo and in vitro. AB - Parkinson's disease is a neurodegenerative disorder of uncertain pathogenesis characterized by a loss of dopaminergic neurons in substantia nigra pars compacta, and can be modeled by the neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6 tetrahydropyridine (MPTP). Oxidative stress may contribute to MPTP- and Parkinson's disease-related neurodegeneration. Fucoidan is a sulfated polysaccharide extracted from brown seaweeds which possesses a wide variety of biological activities including potent antioxidative effects. Here we investigated the effect of fucoidan treatment on locomoter activities of animals, striatal dopamine and its metabolites and survival of nigral dopaminergic neurons in MPTP-induced animal model of Parkinsonism in C57/BL mice in vivo and on the neuronal damage induced by 1-methyl-4-phenylpyridinium (MPP(+)) in vitro, and to study the possible mechanisms. When administered prior to MPTP, fucoidan reduced behavioral deficits, increased striatal dopamine and its metabolites levels, reduced cell death, and led to a marked increase in tyrosine hydroxylase expression relative to mice treated with MPTP alone. Furthermore, we found that fucoidan inhibited MPTP-induced lipid peroxidation and reduction of antioxidant enzyme activity. In addition, pre-treatment with fucoidan significantly protected against MPP(+)-induced damage in MN9D cells. Taken together, these findings suggest that fucoidan has protective effect in MPTP-induced neurotoxicity in this model of Parkinson's disease via its antioxidative activity. PMID- 19545564 TI - Haplorchis pumilio and H. taichui in Vietnam discriminated using ITS-2 DNA sequence data from adults and larvae. AB - Samples of Haplorchis taichui and Haplorchis pumilio of different life-stages (cercariae, metacercariae and adults) and from different host species (snail, fish, dog, cat and human) were collected in Nghe An and Nam Dinh Provinces in Vietnam. Samples from Thailand were available for comparison. All adults and metacercariae were initially identified using morphological criteria. Polymerase chain reaction (PCR) assays were developed for discriminating between the species. The complete sequence for the nuclear ribosomal internal transcribed spacer-2 (ITS-2) was obtained from one adult and one metacercaria of H. taichui and three adults and three metacercariae of H. pumilio from Vietnam. Sequences from cercariae from three different snails clustered with those of H. pumilio. Intra-individual variation in the ITS-2 region was detected by sequencing of cloned PCR products. These are the first sequences from Vietnamese Haplorchis spp. to be reported and demonstrate that H. taichui and H. pumilio can be identified unambiguously from any life-stage, including the cercarial stage that is difficult to identify using morphology. Discrepancies in the literature are discussed and examples of apparent misidentification highlighted. The data provide a resource to assist in taxonomic studies on heterophyids, in the design of probes for diagnosis and for field surveys to identify infection in snails. PMID- 19545565 TI - Host peroxisomal properties are not restored to normal after treatment of visceral leishmaniasis with sodium antimony gluconate. AB - Reason for post-kala-azar dermal leishmaniasis (PKDL) is yet to be established. Earlier it was observed that morphology and biochemical properties of host peroxisomes were impaired during Leishmania infection. As peroxisome is known to be involved in various metabolic pathways to monitor normal function of the host cells, it is essential that Leishmania-induced dysfunction of this organelle should totally be repaired during treatment of visceral leishmaniasis (VL). In this paper it has been shown that resumption of normal peroxisomal function could not be attained when one of the existing drugs sodium antimony gluconate (SAG) was used for chemotherapy against VL. Although Leishmania parasite was found to be completely eliminated from host liver and spleen after SAG treatment, normal activities of peroxisomal catalase and superoxide dismutase could not be restored. Also unusual peptides were found to be present due to abnormal proteolytic cleavage of proteins. It is proposed that peroxisomal disorder which exists even after successful chemotherapy of VL may be figured out as one of the possible reasons to develop PKDL. It may also be pointed out that continued effect of peroxisomal disorder even after complete treatment of this parasitic disease may also lead to genetic disorders not yet been explored in post-kala azar patients. PMID- 19545566 TI - Plasmodium relictum (lineage P-SGS1): Further observation of effects on experimentally infected passeriform birds, with remarks on treatment with Malarone. AB - Plasmodium relictum (lineage P-SGS1) is a widespread malaria parasite that causes disease of different severity in different species of birds. However, experimental studies on the effects of this parasite on avian hosts are uncommon. We investigated development of this lineage in experimentally infected greenfinches Carduelis chloris and compared the obtained data with the literature information about the virulence of the same parasite lineage for phylogenetically closely related bird species. We also used an opportunity to test the efficacy of the antimalarial drug Malarone in treatment of the experimental infection. The cryopreserved strain of the lineage P-SGS1 was multiplied in 4 experimentally infected chaffinches. Light parasitemia developed in these birds; the parasites were then inoculated to 6 uninfected recipient greenfinches. Six uninfected greenfinches were used as negative controls. Light parasitemia developed in all experimental greenfinches. There were no significant effects of malaria on the body mass of greenfinches, but haematocrit value was slightly lower in experimental birds than in control ones; the infection did not cause mortality or morbidity in these birds. According to available data, all investigated fringillid birds are susceptible to P. relictum (P-SGS1), but the same malaria parasite develops markedly differently in different bird species, even closely related hosts. Thus, the observed effects of the same malaria lineage on one species of bird cannot be generalized to others, even closely related ones. The cure with Malarone was highly efficient for blood stages of P. relictum, but exoerythrocytic stages were unaffected. PMID- 19545567 TI - In-depth validation of acridine orange staining for flow cytometric parasite and reticulocyte enumeration in an experimental model using Plasmodium berghei. AB - Flow cytometry is potentially an effective method for counting malaria parasites, but inconsistent results have hampered its routine use in rodent models. A published two-channel method using acridine orange offers clear discrimination between the infected and uninfected erythrocytes. However, preliminary studies showed concerns when dealing with Plasmodium berghei-infected blood samples with high numbers of reticulocytes. In hyperparasitemic or chronic P. berghei infection, enhanced erythropoietic activity results in high numbers of circulating immature reticulocytes. We show that even though the protocol offered good discrimination in newly infected animals, discrimination between infected erythrocytes and uninfected reticulocytes became difficult in animals with hyperparasitemia or chronic infections maintained with subcurative treatment. Discrimination was especially hampered by increased nucleic acid content in immature uninfected reticulocytes. Our data confirms that though flow cytometry is a promising analytical tool in malaria research, care should still be taken when analysing samples from anemic or chronically infected animals. PMID- 19545568 TI - Bridging the gap: from protein misfolding to protein misfolding diseases. AB - Protein misfolding and aggregation are pathognomic for a number of the most common age-related degenerative diseases. Great progress has been made in studying protein aggregation in the test tube and also in replicating protein aggregation in vertebrate animal models of these diseases. However, we argue here that the development and effective integration of emerging techniques such as the methods of nanoscience and the use of invertebrate models are now providing powerful new opportunities to advance our current understanding of the fundamental origins of these disorders. PMID- 19545569 TI - Characterisation of mutations in GroES that allow GroEL to function as a single ring. AB - The chaperonin GroEL contains two seven-subunit rings, and allosteric signals between them are required to complete the GroEL reaction cycle. For this reason SR1, a mutant of GroEL that forms only single rings, cannot function as a chaperone. Mutations in SR1 that restore chaperone function weaken its interaction with the cochaperonin GroES. We predicted that GroES mutants with reduced affinity for GroEL would also restore function to SR1. To test this, we mutated residues in GroES in and near its contact site with GroEL. Nearly half of the mutants showed partial function with SR1. Two mutants were confirmed to have reduced affinity for GroEL. Intriguingly, some GroES mutants were able to function with active single ring mutants of GroEL. PMID- 19545570 TI - The future is wireless: advances in wireless diagnostic and therapeutic technologies in gastroenterology. PMID- 19545571 TI - Glycotope analysis in miracidia and primary sporocysts of Schistosoma mansoni: differential expression during the miracidium-to-sporocyst transformation. AB - Fucosylated carbohydrate epitopes (glycotopes) expressed by larval and adult schistosomes are thought to modulate the host immune response and possibly mediate parasite evasion in intermediate and definitive hosts. While previous studies showed glycotope expression is developmentally and stage-specifically regulated, relatively little is known regarding their occurrence in miracidia and primary sporocysts. In this study, previously defined monoclonal antibodies were used in confocal laser scanning microscopy, standard epifluorescence microscopy and Western blot analyses to investigate the developmental expression of the following glycotopes in miracidia and primary sporocysts of Schistosoma mansoni: GalNAcbeta1-4GlcNAc (LDN), GalNAcbeta1-4(Fucalpha1-3)GlcNAc (LDN-F), Fucalpha1 3GalNAcbeta1-4GlcNAc (F-LDN), Fucalpha1-3GalNAcbeta1-4(Fucalpha1-3)GlcNAc (F-LDN F), GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc (LDN-DF), Fucalpha1-2Fucalpha1 3GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc (DF-LDN-DF), Galbeta1-4(Fucalpha1 3)GlcNAc (Lewis X) and the truncated trimannosyl N-glycan Manalpha1-3(Manalpha1 6)Manbeta1-4GlcNAcbeta1-4GlcNAcbeta1-Asn (TriMan). All but Lewis X were variously expressed by miracidia and sporocysts of S. mansoni. Most notably, alpha3 fucosylated LDN (F-LDN, F-LDN-F, LDN-F) was prominently expressed on the larval surface and amongst glycoproteins released during larval transformation and early sporocyst development, possibly implying a role for these glycotopes in snail schistosome interactions. Interestingly, Fucalpha2Fucalpha3-subsituted LDN (LDN DF, DF-LDN-DF) and LDN-F were heterogeneously surface-expressed on individuals of a given larval population, particularly amongst miracidia. In contrast, LDN and TriMan primarily localised in internal somatic tissues and exhibited only minor surface expression. Immunoblots indicate that glycotopes occur on overlapping but distinct protein sets in both larval stages, further demonstrating the underlying complexity of schistosome glycosylation. Additionally, sharing of specific larval glycotopes with Biomphalaria glabrata suggests an evolutionary convergence of carbohydrate expression between schistosomes and their snail host. PMID- 19545573 TI - Impact of environmental stress on aphid clonal resistance to parasitoids: Role of Hamiltonella defensa bacterial symbiosis in association with a new facultative symbiont of the pea aphid. AB - Resistance to endoparasitoids in aphids involves complex interactions between insect and microbial players. It is now generally accepted that the facultative bacterial symbiont Hamiltonella defensa of the pea aphid Acyrthosiphon pisum is implicated in its resistance to the parasitoid Aphidius ervi. It has also been shown that heat negatively affects pea aphid resistance, suggesting the thermosensitivity of its defensive symbiosis. Here we examined the effects of heat and UV-B on the resistance of A. pisum to A. ervi and we relate its stability under heat stress to different facultative bacterial symbionts hosted by the aphid. For six A. pisum clones harboring four different facultative symbiont associations, the impact of heat and UV-B was measured on their ability to resist A. ervi parasitism under controlled conditions. The results revealed that temperature strongly affected resistance, while UV-B did not. As previously shown, highly resistant A. pisum clones singly infected with H. defensa became more susceptible to parasitism after exposure to heat. Interestingly, clones that were superinfected with H. defensa in association with a newly discovered facultative symbiont, referred to as PAXS (pea aphid X-type symbiont), not only remained highly resistant under heat stress, but also expressed previously unknown, very precocious resistance to A. ervi compared to clones with H. defensa alone. The prevalence of dual symbiosis involving PAXS and H. defensa in local aphid populations suggests its importance in protecting aphid immunity to parasitoids under abiotic stress. PMID- 19545572 TI - A novel nanoparticulate adjuvant for immunotherapy with Lolium perenne. AB - Specific immunotherapy implies certain drawbacks which could be minimized by the use of appropriate adjuvants, capable of amplifying the right immune response with minimal side effects. In this context, previous studies of our group have demonstrated the adjuvant capacity of Gantrez AN nanoparticles, which can effectively enhance the immune response. In this work, two types of nanoparticles (with and without LPS of Brucella ovis as immunomodulator) with encapsulated Lolium perenne extract are tested in a model of sensitized mice to this allergenic mixture. The results we obtained showed that Lolium-Gantrez nanoparticles with LPS of B. ovis were able to induce significative Th1 responses, characterized by the IgG(2a) isotype. Furthermore, in the challenge experiment of the sensitized mice, differences in the mortality rate and in the mMCP-1 levels were found between the treated groups and the control. Under the experimental conditions of this model of pre-sensitized mice to L. perenne, Gantrez AN nanoparticles appeared to be a good strategy for immunotherapy. PMID- 19545574 TI - Physiological and molecular characterization of methotrexate transport by Malpighian tubules of adult Drosophila melanogaster. AB - A radioisotope tracer technique and quantitative PCR were used to study the mechanisms and regulation of transepithelial transport of the type II organic anion methotrexate (MTX) by the Malpighian tubules of Drosophila melanogaster. Transport of MTX was saturable and Na(+)-independent; the kinetic parameters J(max) and K(t) were 437fmolmin(-1) and 23.5microM, respectively. The transport of MTX was competitively inhibited by phenol red and probenecid; non competitively inhibited by salicylate, verapamil and MK-571; and uncompetitively inhibited by Texas Red. Dietary exposure to 0.1mM MTX led to dramatic increases in gene expression for several members of the ABC family of transporters in both the Malpighian tubules and the gut. Our results suggest that multiple transporters are upregulated in response to dietary exposure to MTX. Increased levels of the protein products which may result from expression of these genes may enhance elimination of toxic compounds such as MTX or its metabolites. PMID- 19545575 TI - Disease spread in small-size directed networks: epidemic threshold, correlation between links to and from nodes, and clustering. AB - Network epidemiology has mainly focused on large-scale complex networks. It is unclear whether findings of these investigations also apply to networks of small size. This knowledge gap is of relevance for many biological applications, including meta-communities, plant-pollinator interactions and the spread of the oomycete pathogen Phytophthora ramorum in networks of plant nurseries. Moreover, many small-size biological networks are inherently asymmetrical and thus cannot be realistically modelled with undirected networks. We modelled disease spread and establishment in directed networks of 100 and 500 nodes at four levels of connectance in six network structures (local, small-world, random, one-way, uncorrelated, and two-way scale-free networks). The model was based on the probability of infection persistence in a node and of infection transmission between connected nodes. Regardless of the size of the network, the epidemic threshold did not depend on the starting node of infection but was negatively related to the correlation coefficient between in- and out-degree for all structures, unless networks were sparsely connected. In this case clustering played a significant role. For small-size scale-free directed networks to have a lower epidemic threshold than other network structures, there needs to be a positive correlation between number of links to and from nodes. When this correlation is negative (one-way scale-free networks), the epidemic threshold for small-size networks can be higher than in non-scale-free networks. Clustering does not necessarily have an influence on the epidemic threshold if connectance is kept constant. Analyses of the influence of the clustering on the epidemic threshold in directed networks can also be spurious if they do not consider simultaneously the effect of the correlation coefficient between in- and out degree. PMID- 19545576 TI - Auditory discrimination and auditory sensory behaviours in autism spectrum disorders. AB - It has been hypothesised that auditory processing may be enhanced in autism spectrum disorders (ASD). We tested auditory discrimination ability in 72 adolescents with ASD (39 childhood autism; 33 other ASD) and 57 IQ and age matched controls, assessing their capacity for successful discrimination of the frequency, intensity and duration differences in pairs of sounds. At the group level, auditory discrimination ability did not differ between the adolescents with and without ASD. However, we found a subgroup of 20% of individuals in the ASD group who showed 'exceptional' frequency discrimination skills (defined as 1.65 SDs above the control mean) and who were characterised by average intellectual ability and delayed language onset. Auditory sensory behaviours (i.e. behaviours in response to auditory sensory input) are common in ASD and we hypothesised that these would relate to auditory discrimination ability. For the ASD group, poor performers on the intensity discrimination task reported more auditory sensory behaviours associated with coping with loudness levels. Conversely, those who performed well on the duration discrimination task reported more auditory sensory behaviours across the full range measured. Frequency discrimination ability did not associate with auditory sensory behaviours. We therefore conclude that (i) enhanced frequency discrimination is present in around 1 in 5 individuals with ASD and may represent a specific phenotype; and (ii) individual differences in auditory discrimination ability in ASD may influence the expression of auditory sensory behaviours by modulating the degree to which sounds are detected or missed in the environment. PMID- 19545577 TI - The relationship between awareness and attention: evidence from ERP responses. AB - The relationship between attention and awareness is complex, because both concepts can be understood in different ways. Here we review our recent series of experiments which have tracked the independent contributions of different types of visual attention and awareness to electrophysiological brain responses, and then we report a new experiment focusing on spatial attention, nonspatial selection of objects, and visual consciousness at the same time. The results indicate that the earliest electrophysiological correlate of consciousness, assumed to correlate with "phenomenal consciousness", was dependent on spatial attention, suggesting that spatial attention is a prerequisite for the internal representations of space that provide the medium for phenomenal experience. The correlate of phenomenal consciousness emerged independent of nonspatial selection of objects, but its later part was modified by it. By contrast, the correlate of access to later conscious processing stages ("reflective consciousness") that take the selected contents of phenomenal consciousness as input for conceptual thought and working memory, was dependent on both spatial attention and nonspatial selection. These results imply that one should distinguish between different types of attention and different forms of awareness, when describing the relationship between attention and awareness. PMID- 19545578 TI - Temporal preparation in aging: a functional MRI study. AB - Young and elderly adults performed a choice-RT task while scanned with functional magnetic resonance imaging. A foreperiod separated a warning and a response signal. In the variable condition, the foreperiod varied randomly between 1 and 3s. In the fixed conditions, it was kept constant at either 1 or 3s. Elderly subjects responded slower than controls in both task conditions. An interaction was observed between age and foreperiod in the variable condition only: in the young group, RT decreased with longer foreperiods, whereas the elderly participants showed the opposite tendency. This was accompanied by difference in brain activation. Right lateral prefrontal regions were more activated in the young than in the elderly group in the variable vs. fixed foreperiod contrast. These findings unveil the neural substrate of age-related preparation deficits, and confirm that the involvement of right lateral prefrontal cortex is essential for strategic preparation under uncertain timing conditions. PMID- 19545580 TI - Lesion neuroanatomy of the Sustained Attention to Response task. AB - The Sustained Attention to Response task is a classical neuropsychological test that has been used by many centres to characterize the attentional deficits in traumatic brain injury, ADHD, autism and other disorders. During the SART a random series of digits 1-9 is presented repeatedly and subjects have to respond to each digit (go trial) except the digit '3' (no-go trial). Using voxel-based lesion symptom mapping (VLSM) in a consecutive series of 44 ischemic unifocal non lacunar hemispheric stroke patients we determined the neuroanatomy of 4 SART parameters: commission and omission error rate, reaction time variability and post-error slowing. Lesions of the right inferior frontal gyrus significantly increased commission error rate. Lesions of the middle third of the right inferior frontal sulcus (IFS) reduced post-error slowing, a measure of how well subjects can utilize errors to adjust cognitive resource allocation. Omissions and reaction time variability had less localising value in our sample. To conclude, commission errors and post-error slowing in the SART mainly probe right inferior frontal integrity. PMID- 19545579 TI - Dissociation of decision-making under ambiguity and decision-making under risk in patients with Parkinson's disease: a neuropsychological and psychophysiological study. AB - Decision-making impairments in Parkinson's disease (PD) are most likely associated with dysfunctions in fronto-striatal loops. Recent studies examined decision-making in PD either in ambiguous situations with implicit rules, using the Iowa Gambling Task (IGT), or in risky situations with explicit rules, using the Game of Dice Task (GDT). Both tasks have been associated with the limbic orbitofrontal-striatal loop, involved in emotional processing. However, the GDT has additionally been highly associated with the dorsolateral prefrontal-striatal loop, being involved in executive functions. The present study is the first one which examined decision-making in PD patients with both, IGT and GDT. We studied 21 non-demented PD patients on dopaminergic medication and 23 healthy controls with both tasks and a neuropsychological test battery with focus on executive functions. To analyse possible abnormalities in emotional processing, electrodermal responses (EDRs) were assessed while performing the tasks. We found that PD patients were significantly impaired in the GDT, but not in the IGT. Executive dysfunctions correlated with GDT but not with IGT performance. In both tasks, PD patients showed significantly reduced feedback EDRs after losses, but not after gains, indicating a primary decline of sensitivity to negative feedback. Our behavioural data suggest that dysfunctions in the dorsolateral prefrontal loop might be stronger than in the limbic loop, resulting in deficits in executive functions and GDT performance but unimpaired IGT performance. Reduced sensitivity to negative feedback is discussed with regard to dysfunctions in the limbic loop, which may result from pathology of limbic structures or dopaminergic medication. PMID- 19545581 TI - Individual differences in task-related activation and performance. AB - Previous studies from this laboratory have explored the relationships between physiological response magnitudes, behavioural performance measures, current arousal level, and activation (defined as task-related change in arousal) during a continuous performance task. We have reported larger physiological responses with increased arousal, and performance improvement with higher levels of activation. Performance results have been consistent at the level of averaged data, whether obtained from between-participant studies using across-trials averaging (yielding one data point per person), or using across-subject within trials averaging (yielding 1 data point per trial). The present study was designed to explore this relationship at the individual level. A repeated continuous performance task (CPT) was used with normal adults. Reaction time performance at each target in the first block was considered relative to the corresponding target in the second block. Skin conductance level (SCL) was recorded during the task as an index of arousal at each target. Activation at each target in the first block was calculated by subtracting the level of arousal at the corresponding target in the second block. The relationship between first block performance enhancement and activation in different participants followed different trends. Some participants showed the traditional inverted-U relationship, and some showed the opposite trend. In line with our previous studies, the average linear relationship over all participants was positive. The importance of these findings is discussed in relation to sport performance enhancement. PMID- 19545582 TI - Sexually dimorphic c-Fos expression following spatial working memory in young and adult rats. AB - The sex differences in the functional contribution of brain substrates were explored following acquisition of a spatial working memory task using quantification of c-Fos protein. Rats of both sexes were trained during adolescence and adulthood in Morris water maze using a hidden escape platform with different daily location. Two control groups for each sex and age were added to explore the c-Fos activation not specific to the memory process. These were a free-swimming group (yoked control) and a handled control (CO) group. Behaviorally, no age differences were found in number of days required by males to acquire the task, but females showed a delay in acquisition during adolescence (P30) that improved in adulthood (P90). Both sexes showed a learning-related increase in Fos immunoreactivity in the anterodorsal and anteroventral thalamus and medial and lateral mammillary nuclei during adolescence. Higher levels of learning-related Fos immunoreactivity were found in the infralimbic cortex, CA3 and CA1 only in females. During adulthood the common activated region was the prelimbic cortex with the addition of the infralimbic cortex in the male group and the lateral mammillary nucleus in the female group. These results indicated sex and age differences in brain functioning following working memory task. However, they could not be necessarily linked with differences in performance since similar results were found between males and females during adulthood. The activation of common and interrelated structures suggests that these structures are involved in spatial processing but it also highlights the relevance of developmental changes for understanding the memory process. PMID- 19545583 TI - Eating what you like induces a stronger decrease of 'wanting' to eat. AB - Human eating behavior may be influenced non-homeostatically by the rewarding value of foods, i.e. 'liking' (pleasure/palatability) and 'wanting' (incentive motivation). The objectives of this study were to validate a computer test for assessment of rewarding value of food, and to assess how rewarding value of food is affected by eating a dessert-specific (chocolate mousse, CM) vs. dessert non specific, neutral food item (cottage cheese, CC). Seventy-three subjects (47f/26m, age 27.8+/-10.0 y, BMI 24.1+/-3.3 kg/m(2)), studied in a randomized cross-over design, came to the university twice, fasted. A computer test was developed to determine rewarding value, i.e. 'liking' and 'wanting', for 72 items divided in six categories (bread, filling, drinks, dessert, sweets, stationery). 'Liking' was measured by indicating relative preference of paired items (within/between categories), 'wanting' by working to earn items to choose from. Subjects completed the computer test before and after consumption of CM/CC, matched for energy content (5.6 kJ/g) and daily energy requirements (10%). 'Liking' and 'wanting' scores of all fasted subjects on the two test-days showed 62-73% reproducibility. CM was liked more than CC (p<0.001). Consumption of CM decreased 'wanting' for bread, filling, drinks and dessert (p<0.03). Consumption of CC decreased 'wanting' for bread only (p<0.05). Contrary to CC, CM decreased relative 'liking' for the dessert category (p<0.001). In conclusion, the computer test for measurement of 'liking' and 'wanting' is sufficiently valid. Eating a highly liked food item induces a more distinct decrease in 'wanting' for food items in general and category-specific 'liking', than eating a sufficiently liked neutral food item. PMID- 19545584 TI - Social buffering of the stress response: diversity, mechanisms, and functions. AB - Protracted or repeated activation of the hypothalamic-pituitary-adrenocortical (HPA) system is associated with a variety of physical and psychological pathologies. Studies dating back to the 1970s have documented many cases in which the presence of a social companion can moderate HPA responses to stressors. However, there also are many cases in which this "social buffering" of the HPA axis is not observed. An examination of the literature indicates that the nature of the relationship between individuals is crucial in determining whether or not social buffering of the HPA response will occur. Other factors that affect social buffering, either directly or by influencing the social relationship, include the social organization of the species, previous experience, gender, integration into a social unit, and the developmental stage at which individuals are examined. Current evidence suggests that social buffering involves mechanisms acting at more than one level of the CNS. It is suggested that, in addition to promoting health, social buffering may have evolved to direct the establishment of social relationships, and to facilitate developmental transitions in social interactions appropriate for different life stages. PMID- 19545585 TI - Impulsivity: its genetic, neurochemical and brain substrate determinants and the risks it entails for aberrant motivated behavior and psychopathology. PMID- 19545586 TI - Unraveling the protective effect of a Drosophila phosphatidylethanolamine-binding protein upon bacterial infection by means of proteomics. AB - This study addresses the biological function of CG18594, a Drosophila melanogaster phosphatidylethanolamine-binding protein (PEBP) that we named PEBP1, by combining fly genetics, survival experiments and differential proteomics. We demonstrate that transgenic flies overexpressing PEBP1 are highly protected against bacterial infection due to the release of immunity-related proteins in their hemolymph. Apart from proteins that have been reported earlier to participate in insect immunity, we also identify proteins involved in metabolism and signaling, and, in addition, twelve (hypothetical) proteins with unknown function. This is the first report demonstrating an immune function for a Drosophila PEBP protein. PMID- 19545587 TI - Construction of two gateway vectors for gene expression in fungi. AB - We report the construction of two Gateway fungal expression vectors pCBGW and pGWBF. The pCBGW was generated by introducing an expression cassette, which consists of a Gateway recombinant cassette (attR1-Cmr-ccdB-attR2) under the control of fungal promoter PgpdA and a terminator TtrpC, into the multiple cloning site of fungal vector pCB1004. The pGWBF is a binary vector, which was generated from the plant expression vector pGWB2 by replacing the CaMV35S promoter with PgpdA. The pGWBF can be transformed into fungi efficiently with Agrobacterium-mediated transformation. The applicability of two newly constructed vectors was tested by generating the destination vectors pGWBF-GFP and pCBGW-GFP and examining the expression of GFP gene in Trichoderma viride and Gibberella fujikuroi, respectively. Combining with the advantage of Gateway cloning technology, pCBGW and pGWBF will be useful in fungi for large-scale investigation of gene functions by constructing the interested gene destination/expression vectors in a high-throughput way. PMID- 19545589 TI - Pluripotency rush! Molecular cues for pluripotency, genetic reprogramming of adult stem cells, and widely multipotent adult cells. AB - In the last few years, pluripotent stem cells have been the objective of intense investigation efforts. These cells are of paramount therapeutic interest, since they could be utilized: as in vitro models of disease, for pharmaceutical screening purposes, and for the regeneration of damaged organs. Over the years, pluripotent cells have been cultured from teratomas, the inner cell mass, and primordial germ cells. Accumulating informations have partially decrypted the molecular machinery responsible for the maintenance of a very primitive state, permitting the reprogramming of differentiated cells. Although the debate is still open, an extreme excitement is arising from two strictly related possibilities: pluripotent cells could be obtained from adult tissues with minimal manipulations or very rare pluripotent cells could be identified in adult tissues. This intriguing option will trigger new researches aimed both at identifying the possible biological role of pluripotent adult stem cells and at exploiting their potential clinical use. The present review article will summarize current knowledge of the molecular cues for pluripotency but also discusses whether pluripotent stem cells could be obtained from adult tissues. PMID- 19545590 TI - Novel therapeutics for type 2 diabetes: incretin hormone mimetics (glucagon-like peptide-1 receptor agonists) and dipeptidyl peptidase-4 inhibitors. AB - Known treatments of type 2 diabetes mellitus have limitations such as weight gain, and hypoglycaemias. A new perspective is the use of incretin hormones and incretin enhancers. Incretins are defined as being responsible for the higher insulin release after an oral glucose load compared to an intravenous glucose load. The delicate balance of glucose homeostasis, in which incretin hormones are involved, is disturbed in type 2 diabetes mellitus. The incretin GLP-1 helps to maintain glucose homeostasis through stimulation of insulin secretion and inhibition of glucagon release in a glucose-dependent manner. This is associated with reductions in body weight, and no risk of hypoglycaemias. When classical oral agents have failed to maintain adequate glycaemic control, incretin mimetics may be of particular value for obese patients and those who have little control over meal sizes. Exenatide was marketed as a GLP-1 analogue and longer acting incretin mimetics such as liraglutide, albiglutide and others have the same pharmacological profile. In addition to incretin mimetics incretin enhancers which inhibit/delay degradation of incretins were developed: so-called DPP-4 inhibitors such as sitagliptin and vildagliptin are approved in Europe. Their differences from incretin mimetics include: oral bioavailability, less side effects with overdose, no direct CNS effects (nausea and vomiting) and no effect on weight. In rodent models of diabetes, but not yet in humans, GLP-1 receptor agonists and DPP-4 inhibitors increase islet mass and preserve beta-cell function. Incretin mimetics and enhancers expand type 2 diabetes treatment, are still not first line therapy and it is discussed if they are to be prophylactically used. PMID- 19545591 TI - The Trypanosoma brucei sphingolipid synthase, an essential enzyme and drug target. AB - Sphingolipids are important components of eukaryotic membranes, particularly the plasma membrane, and are involved in a diverse array of signal transduction processes. In the Eukaryota the biosynthetic pathway for the formation of these lipid species is largely conserved. However, in contrast to mammals which produce sphingomyelin (SM), several pathogenic fungi and protozoa synthesize inositol phosphorylceramide (IPC) as the primary phosphosphingolipid. This process is catalyzed by the enzyme IPC synthase, a recognized target for anti-fungals encoded by the AUR1 gene in yeast. Recently, functional orthologues of the AUR1p have been identified in a group of insect vector-borne pathogenic protozoa, the Kinetoplastida, which are responsible for a range of so-called neglected diseases. Of these the Trypanosoma brucei species are the causative agents of human African trypanosomiasis in many of the most under-developed regions of Africa. The available treatments for these diseases are limited, of decreasing efficacy, and often demonstrate severe side-effects. Against this background the T. brucei sphingolipid synthase, an orthologue of the yeast AUR1p, may represent a promising target for novel anti-protozoals. Our studies identify an isoform of this protein as a novel bi-functional enzyme capable of catalyzing the synthesis of both IPC and SM, both known to be present in the parasite. Furthermore, the synthase is essential for parasite growth and can be inhibited by a known anti fungal at low nanomolar levels in vitro. Most notably this drug demonstrates trypanocidal activity against cultured bloodstream form parasites. Thus, the T. brucei sphingolipid synthase represents a valid and promising drug target. PMID- 19545593 TI - Treatment of osteomyelitis and repair of bone defect by degradable bioactive borate glass releasing vancomycin. AB - The effectiveness of a degradable and bioactive borate glass has been compared with the clinically used calcium sulfate in the treatment of osteomyelitis of rabbits, as a carrier for vancomycin. The bone infections were induced in the tibias of 65 rabbits by injecting methicillin-resistant Staphylococcus aureus (MRSA). After 3 weeks, these rabbits were distributed into 4 groups and treated by debridement. Pure borate glass (BG), vancomycin-loaded calcium sulfate (VCS) and vancomycin-loaded borate glass (VBG) were implanted into the infection sites of groups 2 to 4 respectively. After 8 weeks, the effectiveness of treatment was assessed radiographically, bacteriologically, and histopathologically. The results showed that the negative rates of MRSA examination for rabbits were 36.36%, 18.18%, 73.33% and 81.25% respectively for groups 1 to 4. Significant differences were observed radiographically, bacteriologically, and histopathologically between groups 1 and 4, groups 2 and 3, and between groups 2 and 4. The best result of treatment was observed in group 4. Radiographically, VBG was found to be mostly reabsorbed and replaced by lots of new bones, whereas, VCS was completely reabsorbed and replaced by modest new bones. Histopathologically, there were lots of newly formed bones around VBG without any foreign body response, and only modest new bones around VCS with obvious foreign body response. VBG proved to have excellent biocompatibility and to be very effective in eradicating osteomyelitis and simultaneously stimulating bone regeneration, avoiding the disadvantages of VCS. PMID- 19545588 TI - Prenatal alcohol exposure: fetal programming and later life vulnerability to stress, depression and anxiety disorders. AB - Children with fetal alcohol spectrum disorder (FASD) exhibit cognitive, neuropsychological and behavioral problems, and numerous secondary disabilities including depression and anxiety disorders. Dysregulation of the hypothalamic pituitary-adrenal (HPA) axis is common in depression/anxiety, reflected primarily in increased HPA tone or activity. Prenatal alcohol exposure (PAE) increases HPA tone and results in HPA dysregulation throughout life, paralleling many of the HPA changes in depression/anxiety. We review data demonstrating altered HPA function and increased depression/anxiety in FASD. In the context of the stress diathesis model, we discuss the hypothesis that fetal programming of the HPA axis by PAE alters neuroadaptive mechanisms that mediate the stress response, thus sensitizing the organism to stressors encountered later in life, and mediating, at least partly, the increased vulnerability to depression/anxiety disorders. Furthermore, we present evidence demonstrating sex-specific alterations in both hormonal and behavioral responsiveness to tasks measuring depressive- and anxiety like behaviors in PAE offspring. Overall, the research suggests that the stress diathesis model provides a powerful approach for elucidating mechanisms underlying the increased vulnerability to mental illness among individuals with FASD, and developing appropriate treatments for these individuals. Dr. Seymour Levine's seminal work on the long-term consequences of early life experiences formed a framework for the development of the research described in this review. PMID- 19545592 TI - Investigation of a dopamine receptor in Schistosoma mansoni: functional studies and immunolocalization. AB - A dopamine receptor (SmD2) was cloned from adult Schistosoma mansoni. The receptor has the classical heptahelical topology of class A (rhodopsin-like) G protein-coupled receptors (GPCR) and shares sequence homology with D2-like receptors from other species. The full length SmD2 cDNA was expressed in the yeast Saccharomyces cerevisiae and mammalian HEK293 cells. Functional assays in both expression systems revealed that SmD2 was responsive to dopamine in a dose dependent manner, whereas other structurally related amines had no effect. Activation of SmD2 in mammalian cells caused an elevation in intracellular cAMP but not calcium, suggesting that the receptor coupled to Gs and the stimulation of adenylate cyclase. Pharmacological studies showed that the S. mansoni dopamine receptor was inhibited by apomorphine, a classical dopamine agonist, as well as known dopaminergic antagonists, including chlorpromazine, spiperone and haloperidol. SmD2 immunoreactivity was detected in membrane protein fractions of S. mansoni cercaria, in vitro transformed schistosomula and adult parasites, using a specific peptide antibody. When tested by confocal immunofluorescence, SmD2 was detected in the subtegumental somatic musculature and acetabulum of all larval stages tested. In the adults, SmD2 was enriched in the somatic muscles and, to a lesser extent, the muscular lining of the caecum. The results suggest that SmD2 is an important component of the neuromuscular system in schistosomes. PMID- 19545594 TI - Protein kinase C alpha-specific peptide substrate graft-type copolymer for cancer cell-specific gene regulation systems. AB - We recently proposed a novel gene regulation system responding to specifically and abnormally activated intracellular enzymes in diseased cells. In the present study, we focused on protein kinase C (PKC)alpha, which is hyper-activated in most tumor cells, as a trigger for transgene regulation. We prepared cationic copolymers comprising hydrophilic and neutral polymers in main chains and cationic peptide substrates with different contents in side chains. Our copolymer with high peptide content (>3 mol%) condensed with pDNA more weakly than with poly(L-lysine) (pLL) having a similar molecular weight, but gene suppression was nearly identical to that of pLL, probably due to the steric hindrance of the main chains in our copolymer. Steric hindrance of the main chains barely affected the phosphorylation reaction of the pendant peptide. In cell and mouse experiments, higher gene expression was observed in complexes of pDNA with copolymers pended PKC alpha-specific substrate peptide than that in complexes with negative copolymers pended peptide substituted phosphorylation site of serine residues with alanine. These results indicate that our system can recognize intracellular PKC alpha as a trigger to regulate transgene expression, and may be useful for tumor gene therapy. PMID- 19545595 TI - A modified protocol for efficient DNA encapsulation into pegylated immunoliposomes (PILs). AB - Effective delivery of transgenes to the brain through a non-invasive route has great prospects for treating diseases in the central nervous system (CNS). Slightly anionic pegylated immunoliposomes (PILs) have been shown to be effective in reaching the CNS, but efficient DNA encapsulation into the liposomes used for this purpose is technically difficult and hard to reproduce. We here use an improved protocol for DNA encapsulation of pegylated immunoliposomes based on ethanol-mediated DNA condensation. We introduce a dialysis step following DNA encapsulation to remove ethanol and show that this step is necessary to ensure complete nucleolytic removal of non-encapsulated DNA. The uptake of the pegylated immunoliposomes into human cells was documented by live-cell confocal imaging, and specific targeting to the human insulin receptor was shown by inhibiting clathrin-mediated endocytosis. PMID- 19545597 TI - Phosphorylation of Bcl-2 and activation of caspase-3 via the c-Jun N-terminal kinase pathway in ursolic acid-induced DU145 cells apoptosis. AB - There is currently no successful therapy for androgen-independent prostate cancer. Ursolic acid (UA), a pentacyclic triterpenoid compound, has been shown to have an anti-proliferative effect on various tumors. We investigated the effect of UA on cell viability in the human hormone-refractory prostate cancer cell line DU145, as well as the molecular mechanisms underlying its growth inhibiting effect. We demonstrated that UA induces apoptosis and the activation of caspase-3 in DU145 cells. UA also causes the activation of c-Jun N-terminal kinase (JNK), but has no effect on extracellular signal-regulated protein kinases (ERK1/2) and p38 MAP kinases (p38). UA-induced JNK activation could result in Bcl-2 phosphorylation (Ser70) and degradation in DU145 cells, which may be one of the molecular mechanisms by which it induces apoptosis. Although further evaluation, such as in vivo testing, is clearly needed, the present results suggest the potential utility of UA as a novel therapeutic agent in advanced prostate cancer. PMID- 19545596 TI - Gingival vascular damage in atherosclerotic rabbits: hydroxytyrosol and squalene benefits. AB - Cardiovascular alterations and periodontal disease have been associated, although cardiovascular disease treatments have not yet been tested against periodontal alterations. We investigated effects of squalene, hydroxytyrosol and coenzyme Q(10) on gingival tissues of rabbits fed on an atherosclerotic diet. Forty-eight rabbits were distributed in six groups. Control group was fed on standard chow for 80 days. The rest were fed with an atherogenic diet for 50 days. After that, a group was sacrificed and the rest were subjected for another extra 30 days on commercial chow alone or supplemented with coenzyme Q(10), squalene or hydroxytyrosol. Atherosclerotic rabbits had higher fibrosis and endothelial activation and lower cellularity in gingival mucosa than controls (P<0.05). Hydroxytyrosol reduced endothelial activation (P<0.05) and squalene additionally decreased fibrosis (P<0.05). Results suggest that gingival vascular changes after the atherosclerotic diet have been reversed by hydroxytyrosol and squalene, natural products from the minor fraction of virgin olive oil. PMID- 19545598 TI - Cycloheximide-induced inhibition of protein synthesis in hippocampal pyramidal neurons is time-dependent: differences between CA1 and CA3 areas. AB - Cycloheximide (CHI), an inhibitor of protein synthesis, is widely used for studying the mechanisms of consolidation of long-term memory (LTM). High concentrations of CHI inhibit the protein synthesis in brain homogenates by more than 80% and impair LTM consolidation. For understanding the mechanisms of consolidation, it is important to know how protein synthesis inhibitors affect hippocampal neurons. However, the effect of CHI on protein synthesis in CA1 and CA3 hippocampal pyramidal neurons is still poorly understood. In the present work, the state of ribosomes in CA1 and CA3 pyramidal neurons from the dorsal hippocampus of Wistar rats 1, 2, 4, and 72 h after the introcerebroventricular (i.c.v.) injection of a high concentration of CHI was determined using the fluorescent dye acridine orange. We showed that CHI induces great differences in the dynamics of the intensity of protein synthesis in CA1 and CA3 pyramidal neurons. The suppression of the intensity of protein synthesis in CA1 pyramidal neurons 1h after the injection of CHI was more than threefold stronger than in CA3, and by 4h, it was most pronounced in CA3 neurons. We suggest that the protein synthesis in CA1 pyramidal neurons contributes significantly to the synaptic consolidation of declarative memory in the first critical period. PMID- 19545599 TI - Immobility in the tail suspension test predicts quinine but not saccharin intake in mice. AB - It is assumed that depressive symptomatology can alter taste preferences in humans. The aim of the present study was to search for correlations between immobility in the tail suspension test (TST) and consumption of saccharin (0.0025 0.1%, w/w) and quinine (0.0024-0.04%) solutions. Male C57BL/6J mice were divided into high immobility and low immobility groups based on their immobility scores in the TST. The groups consumed similar amounts of saccharin solutions in the two bottle choice test. There were significant differences between the groups in quinine intake and preference. Intake of, and preference for, 0.0024% quinine was significantly higher in the high immobility than in low immobility subjects. In line with some animal and human studies, our results suggest that behavioral despair in the TST can correlate with taste responses to bitter stimuli. PMID- 19545600 TI - Influence of perceived egocentric coordinates on the subjective visual vertical. AB - The majority of previous studies which have explored the mechanisms underlying perception of the direction of gravity in static roll tilt have proposed that the tendency to estimate the subjective visual vertical (SVV) as tilted towards body tilt ('Aubert effect') arises from an underestimation of perceived body tilt. The present study has evaluated an alternative assumption that erroneous estimates of verticality may be related to the ability to estimate the orientation of external objects with respect to the observer's perceived body Z-axis. Experiments showed that Aubert effects and the overestimation of 30-90 degrees angles from the body Z-axis in the roll plane were both related to errors made in adjusting a visual rod parallel to the body Z-axis. The results suggest that errors in providing visual estimates of the observer's own body Z-axis reference are implicated in Aubert effect. PMID- 19545601 TI - How many people are able to control a P300-based brain-computer interface (BCI)? AB - An EEG-based brain-computer system can be used to control external devices such as computers, wheelchairs or Virtual Environments. One of the most important applications is a spelling device to aid severely disabled individuals with communication, for example people disabled by amyotrophic lateral sclerosis (ALS). P300-based BCI systems are optimal for spelling characters with high speed and accuracy, as compared to other BCI paradigms such as motor imagery. In this study, 100 subjects tested a P300-based BCI system to spell a 5-character word with only 5 min of training. EEG data were acquired while the subject looked at a 36-character matrix to spell the word WATER. Two different versions of the P300 speller were used: (i) the row/column speller (RC) that flashes an entire column or row of characters and (ii) a single character speller (SC) that flashes each character individually. The subjects were free to decide which version to test. Nineteen subjects opted to test both versions. The BCI system classifier was trained on the data collected for the word WATER. During the real-time phase of the experiment, the subject spelled the word LUCAS, and was provided with the classifier selection accuracy after each of the five letters. Additionally, subjects filled out a questionnaire about age, sex, education, sleep duration, working duration, cigarette consumption, coffee consumption, and level of disturbance that the flashing characters produced. 72.8% (N=81) of the subjects were able to spell with 100% accuracy in the RC paradigm and 55.3% (N=38) of the subjects spelled with 100% accuracy in the SC paradigm. Less than 3% of the subjects did not spell any character correctly. People who slept less than 8h performed significantly better than other subjects. Sex, education, working duration, and cigarette and coffee consumption were not statistically related to differences in accuracy. The disturbance of the flashing characters was rated with a median score of 1 on a scale from 1 to 5 (1, not disturbing; 5, highly disturbing). This study shows that high spelling accuracy can be achieved with the P300 BCI system using approximately 5 min of training data for a large number of non-disabled subjects, and that the RC paradigm is superior to the SC paradigm. 89% of the 81 RC subjects were able to spell with accuracy 80-100%. A similar study using a motor imagery BCI with 99 subjects showed that only 19% of the subjects were able to achieve accuracy of 80-100%. These large differences in accuracy suggest that with limited amounts of training data the P300-based BCI is superior to the motor imagery BCI. Overall, these results are very encouraging and a similar study should be conducted with subjects who have ALS to determine if their accuracy levels are similar. PMID- 19545602 TI - Temporal distribution of mRNA expression levels of various genes in the developing human inferior colliculus. AB - The inferior colliculus (IC) is a major binaural integration center in the auditory pathway. Interestingly, studies on the prenatal development of the human IC are lacking. During development of the nervous system a large repertoire of proteins is involved in transforming simple neuroblast cells into functional elements of the adult neural circuits. The present study reports on the mRNA levels produced by 12 genes involved in pre- [12-29 weeks of gestation (WG)], postnatal [40 postnatal days (PND) as well as 2 and 5 postnatal months (PNM)] developing human IC. The mRNA expression levels of nestin, vimentin, GFAP and DCX during 12-24 WG indicate the stages of neurogenesis, migration and differentiation of the human fetal IC. A decrease in the GAP-43 mRNA levels along with an increase in synaptophysin and PSD-95 mRNA levels during late gestational ages (24-29 WG) suggests the formation of primitive contacts by neurons with their targets and the onset of synapse formation. Expression levels of EGAD mRNA were transient with an increase in the early gestational ages, whereas that of GAD-67 mRNA increased in late gestational ages, indicating the changing role of GABA from a trophic factor to that of a neurotransmitter. High levels of BDNF, NT 3 and MBP mRNA in the late gestational ages reveal that the human IC undergoes neuronal maturation, synaptogenesis and myelination by 29 WG. Therefore, it may be suggested that the morphological maturation of the human IC occurs between 22 and 29 WG and that this period appears to be critical in the shaping of adult like physiological attributes. PMID- 19545603 TI - What are the roles of microRNAs at the mammalian synapse? AB - The modification of neuronal connections in response to stimuli is believed to be the basis of long-term memory formation. It is currently accepted that local protein synthesis critically contributes to site-restricted modulation of individual synapses. Here, we summarize recent evidence implicating miRNAs in this process, leading to altered dendrite morphogenesis and synaptic plasticity. Second, we discuss findings in non-neuronal systems about how RNA-binding proteins can modulate miRNA-mRNA interactions, and how these mechanisms might apply to neurons. Finally, we review recent findings that P-bodies may be important sites for miRNA action at the synapse. PMID- 19545604 TI - Translocator protein (TSPO 18kDa) is expressed by neural stem and neuronal precursor cells. AB - Translocator protein 18 kDa, the peripheral benzodiazepine receptor by its earlier name, is a mitochondrial membrane protein associated with the mitochondrial permeability pore. While the function of the protein is not properly understood, it is known to play roles in necrotic and apoptotic processes of the neural tissue. In the healthy adult brain, TSPO expression is restricted to glial cells. In developing or damaged neural regions, however, TSPO appears in differentiating/regenerating neurons. Using immunocytochemical, molecular biological and cell biological techniques, we demonstrate that TSPO mRNA and protein, while missing from mature neurons, are present in neural stem cells and also in postmitotic neuronal precursors. Investigating some distinct stages of in vitro differentiation of NE-4C neural stem cells, TSPO 18 kDa was found to be repressed in a relatively late phase of neuron formation, when mature neuron-specific features appear. This timing indicates that mitochondria in fully developed neurons display specific characteristics and provides an additional marker for characterising neuronal differentiation. PMID- 19545606 TI - The utility of brief, spectrally rich, dynamic sounds in the passive oddball paradigm. AB - Experiments investigating auditory processing often utilize spectrally rich, dynamic stimuli to simulate an ecologically valid auditory environment in the laboratory. Often, however, these stimuli do not allow for a strict control of the timing of auditory sensory information which may be distributed over the whole duration of a given sound. In the present study, brief (20 ms long), dynamic, spectrally rich sounds were presented in the context of a passive oddball paradigm to young adults. The short duration made certain that the sensory information was delivered entirely within a 20 ms interval. Two sounds were presented as standards (45-45% probability), other two as deviants (5-5% probability) in random sequences, with a stimulus onset asynchrony (SOA) of 1500 ms. Deviants elicited the mismatch negativity and late difference negativity (LDN) event-related potential components. No N1-effect was produced by deviants, which suggests that the acoustic energy is spread over many different features due to the dynamic spectral properties, which, combined with the brief duration, causes insignificant refractoriness-effects at the present SOA. These results support the usefulness of brief natural sounds in auditory research. The elicitation of LDN in an adult group was an unexpected finding, because LDN is mostly found in children, but not in adults. This result might indicate that LDN elicitation depends on stimulation complexity: stimulus settings in which an LDN is registered in children but not in adults may be perceived as more complex by children than by adults. PMID- 19545605 TI - Inhalative formaldehyde exposure enhances aggressive behavior and disturbs monoamines in frontal cortex synaptosome of male rats. AB - Formaldehyde (FA) exposure is known to be toxic to central nervous system of mammals. In this paper, we evaluated the aggressive behavior after repetitive inhalative FA exposure to male SD rats, and explored the potential mechanism. The rats, ranging from 160 to 180 g, were randomly designated into the orchiectomized (ORX) group, the control and the inhalative FA treatment groups. Eight rats underwent orchiectomy surgery. Three days after the orchiectomy surgery, the inhalative FA (monitored to be 13.5+/-1.5 ppm) treatment began. We found that the male SD rats, those were exposed to FA showed more aggressive behavior compared to the control. And the ORX rats exhibited less aggressive behavior than the control. Furthermore, the dopamine increased and 5-HT decreased significantly in frontal cortex synaptosome after inhalative FA treatment. It is the first to evaluate aggressive behavior and identified monoamines disturbances in the frontal cortex synaptosome after the repetitive inhalative FA exposure to rodents. PMID- 19545608 TI - The modulatory effect of substance P on rat pineal norepinephrine release and melatonin secretion. AB - Secretion of melatonin by the mammalian pineal gland is primarily regulated by the release of norepinephrine (NE) from sympathetic nerve terminals that originate from the superior cervical ganglia. Peptidergic nerves that originate in the perikarya located in the sensory trigeminal ganglia also innervate the pineal gland. Some of these peptidergic nerve fibers contain substance P. Previously, we have characterized neurokinin 1 type substance P receptors in the pineal gland. However, the function of this receptor in the pineal gland remains unclear. Here, we examined the modulatory effect of substance P on rat pineal NE transmission. We show that at the presynaptic level, substance P stimulates the KCl-induced [(3)H]NE release from the pineal nerve ending. However, we found that substance P did not affect the basal levels of either arylalkylamine-N acetyltransferase (AANAT) activity or melatonin secretion in rat pineal organ cultures. However, in the presence of NE, substance P inhibited the NE-induced increase in AANAT activity and melatonin secretion. This is the first time that a function for substance P in the mammalian pineal gland has been demonstrated. PMID- 19545607 TI - Mustard oil enhances spinal neuronal responses to noxious heat but not cooling. AB - The TRPA1 agonist mustard oil (allyl isothiocyanate=AITC) induces heat hyperalgesia and mechanical allodynia in human skin and sensitizes rat spinal wide dynamic range (WDR) neuronal responses to noxious skin heating. We presently used electrophysiological methods to investigate if AITC affects the responsiveness of individual spinal WDR neurons to intense skin cooling. Recordings were made from cold-sensitive WDR neurons in lamina I and deeper dorsal horn; 21/23 also responded to noxious skin heating. Topical application of AITC excited 8/18 units and significantly enhanced their responses to noxious heat while not significantly affecting responses to the cold stimulus. Vehicle (mineral oil) had no effect on thermal responses. The data confirm a role for the TRPA1 agonist AITC in enhancing heat nociception without significantly affecting cold sensitivity. PMID- 19545609 TI - Increase in corticospinal excitability of limb and trunk muscles according to maintenance of neck flexion. AB - The effect of maintenance of neck flexion on corticospinal excitability of limb and trunk muscles was investigated using transcranial magnetic stimulation (TMS). Nine healthy young subjects participated in this experiment. Every measurement was performed with subjects sitting on a chair. Target muscles were the first dorsal interosseous (FDI), biceps brachii (BB), triceps brachii (TB), rectus abdominis (RA), erector spinae (ES), rectus femoris (RF), biceps femoris (BF), tibialis anterior (TA), and gastrocnemius (GcM) on the right side. TMS was applied to the left primary motor cortex, and motor evoked potential (MEP) was measured from the muscles listed above. Optimal stimulus location and resting motor threshold (RMT) were identified for each target muscle, and stimulus intensity used was 120% of RMT. MEPs of the target muscle were recorded with the chin resting on a chin support (chin-on condition) with neck in 20 degrees of flexion, and with voluntary maintenance of the neck flexion posture (chin-off condition). Amplitude and latency of MEP and background activity of target muscles were analyzed. For FDI, BB, TB, ES, and RF, amplitude of MEP increased and latency shortened in the chin-off compared with the chin-on condition. No significant difference in background activity of each target muscle was found between the two conditions. Corticospinal excitability of limb and trunk muscles was selectively enhanced while neck flexion was maintained. PMID- 19545610 TI - Brain derived neurotrophic factor and trk B mRNA expression in the brain of a brain stem-spinal cord regenerating model, the European eel, after spinal cord injury. AB - Evidence from mammalian studies suggests that brain derived neurotrophic factor (BDNF) and its receptor, trk B, are upregulated in neuronal cell bodies after injury. Although fish possess neurotrophins and display rapid functional and morphological recovery after central nervous system (CNS) injury, to date few studies have examined neurotrophin expression during CNS regeneration. In this study, RT-PCR was used to investigate the effect of complete spinal cord transection on the mRNA expression of BDNF and its receptor, trk B, in the eel brain at a range of timepoints after injury. The spatial expression pattern of BDNF mRNA in the brain was also assessed before and after injury using in situ hybridization. Marked changes in BDNF and trk B mRNA levels in the eel brain were not detected during the recovery period after cord transection. In addition, the spatial expression pattern of BDNF mRNA in the eel brain appeared unchanged after injury. Our results are in contrast with the increase reported in mammals but are in line with studies examining neurotrophin expression during CNS regeneration in other anamniotic vertebrates. PMID- 19545611 TI - Edaravone, a free radical scavenger, retards the development of amygdala kindling in rats. AB - This study evaluated the antiepileptogenic effects of edaravone, a newly developed radical scavenger, on the amygdala kindling rats. The afterdischarge duration (ADD), AD threshold (ADT), and seizure severity in animals were measured to study the anticonvulsant effects of edaravone (2mg/kg or 20mg/kg i.p. for 7 days) on fully kindled seizures. Furthermore, for the study of antiepileptogenesis effects of the drug (2mg/kg or 20mg/kg i.p. for 7 days), not only ADD and seizure severity during kindling but also both the pre- and post kindling ADT were measured. Edaravone neither induces nor inhibits fully kindled seizures regardless of the dose; however high-dose edaravone (20mg/kg) retarded kindling development together with shortened ADD and elevated ADT. The present data suggest that high-dose edaravone has an antiepileptogenic drug effect for the prevention of epilepsy. However, other chronic models and clinical trials are needed to confirm the effects of edaravone on the prevention of human epilepsy. PMID- 19545612 TI - Worry tendencies predict brain activation during aversive imagery. AB - Because of its abstract nature, worrying might function as an avoidance response in order to cognitively disengage from fearful imagery. The present functional magnetic resonance imaging study investigated neural correlates of aversive imagery and their association with worry tendencies, as measured by the Penn State Worry Questionnaire (PSWQ). Nineteen healthy women first viewed, and subsequently imagined pictures from two categories, 'threat' and 'happiness'. Worry tendencies were negatively correlated with brain activation in the anterior cingulate cortex, the prefrontal cortex (dorsolateral, dorsomedial, ventrolateral), the parietal cortex and the insula. These negative correlations between PSWQ scores and localized brain activation were specific for aversive imagery. Moreover, activation in the above mentioned regions was positively associated with the experienced vividness of both pleasant and unpleasant mental pictures. As the identified brain regions are involved in emotion regulation, vivid imagery and memory retrieval, a lowered activity in high PSWQ scorers might be associated with cognitive disengagement from aversive imagery as well as insufficient refresh rates of mental pictures. Our preliminary findings encourage future imagery studies on generalized anxiety disorder patients, as one of the main symptoms of this disorder is excessive worrying. PMID- 19545613 TI - The effects of unilateral medial arch support stimulation on plantar pressure and center of pressure adjustment in young gymnasts. AB - The purpose of this study was to examine the contribution of tactile afferents from the medial arch of the foot on postural control. The center of pressure (CoP) position and right/left plantar pressure distributions of 13 gymnasts, with and without a medial arch support, were recorded by a force platform coupled with a baropedometry analysis. Stimulation of the subject's plantar sole was accomplished using a 3mm thick medial arch insert. Right arch stimulation induced an ipsilateral increase of plantar pressure and a contralateral displacement of the CoP to the left. Left arch support also resulted in an ipsilateral increase in plantar pressure and displacement of the CoP to the right. Stimulation of the plantar arch may induce a perception that the body's center of mass has shifted toward the stimulated foot. To maintain stability, individuals may then shift their CoP in the opposite direction. This response may involve compensatory muscle activation strategies to adjust posture. Clinicians may apply these results in their use of foot orthoses to address postural anomalies in patients. PMID- 19545614 TI - Lateralized effects of hand and eye on anticipatory postural adjustments in visually guided aiming movements. AB - This study investigated hemisphere-specific processing of visually aimed movements and associated postural adjustments while controlling for handedness and eyedness. Eleven right-handed, right-eyed and right-footed healthy adult volunteers performed, from a standing position, an aiming task under two hand (right and left hand) and three visual conditions (binocular vision, right and left eye monocular vision). Centre of pressure (CoP) displacement, hand kinematics and the target's position were synchronously recorded during performance of the aiming task. Analysis revealed a lower RMS error, a later postural adjustment onset and a smaller centre of pressure dispersion when aiming was performed with the dominant right compared to the non-dominant left hand. On the other hand, no differences on either aiming performance or postural adjustments were noted under the three visual conditions. These results suggest a strong handedness and absence of an eyedness effect on the accuracy of aiming and associated postural adjustments. PMID- 19545615 TI - In vivo hypoglycemic effects of phenolics from the root bark of Morus alba. AB - Moracin M (1), Steppogenin-4'-O-beta-D-glucosiade (2), Mullberroside A (3) were isolated from the root bark of Morus alba L. and identified by spectral evidence. Compounds 1, 2 and 3 were studied in hypoglycemic effects on alloxan-diabetic mice. The results showed that compounds 1, 2 and 3 all produced hypoglycemic effects. The compound 2 in a dose of 50 mg/kg exerted significant effect (p<0.05), 2 and 3 in a dose of 100 mg/kg exerted obviously effect (p<0.01). Meantime, the compound 1 in a dose of 100 mg/kg can make the fasting blood glucose level have decreasing tendency. PMID- 19545616 TI - Biopharmaceutical characterisation of insulin and recombinant human growth hormone loaded lipid submicron particles produced by supercritical gas micro atomisation. AB - Homogeneous dispersions of insulin and recombinant human growth hormone (rh-GH) in tristearin/phosphatidylcholine/PEG mixtures (1.3:1.3:0.25:0.15 w/w ratio) were processed by supercritical carbon dioxide gas micro-atomisation to produce protein-loaded lipid particles. The process yielded spherical particles, with a 197+/-94 nm mean diameter, and the insulin and rh-GH recovery in the final product was 57+/-8% and 48+/-5%, respectively. In vitro, the proteins were slowly released for about 70-80 h according to a diffusive mechanism. In vivo, the insulin and glucose profiles in plasma obtained by subcutaneous administration of a dose of particles containing 2 microg insulin to diabetic mice overlapped that obtained with 2 microg of insulin in solution. Administration of a dose of particles containing 5 microg insulin resulted in faster and longer glycaemia reduction. Oral administration of 20 and 50 microg insulin equivalent particles produced a significant hypoglycaemic effect. The glucose levels decreased since 2h after administration, reaching about 50% and 70% glucose reduction in 1-2h with the lower and higher dose, respectively. As compared to subcutaneous administration, the relative pharmacological bioavailability obtained with 20 and 50 microg equivalent insulin particles was 7.7% and 6.7%, respectively. Daily subcutaneous administration of 40 microg of rh-GH-loaded particles to hypophysectomised rats induced similar body weight increase as 40 microg rh-GH in solution. The daily oral administration of 400 microg rh-GH equivalent particles elicited a slight body weight increase, which corresponded to a relative pharmacological bioavailability of 3.4% compared to subcutaneous administration. PMID- 19545617 TI - Neural activity underlying tinnitus generation: results from PET and fMRI. AB - Tinnitus is the percept of sound that is not related to an acoustic source outside the body. For many forms of tinnitus, mechanisms in the central nervous system are believed to play an important role in the pathology. Specifically, three mechanisms have been proposed to underlie tinnitus: (1) changes in the level of spontaneous neural activity in the central auditory system, (2) changes in the temporal pattern of neural activity, and (3) reorganization of tonotopic maps. The neuroimaging methods fMRI and PET measure signals that presumably reflect the firing rates of multiple neurons and are assumed to be sensitive to changes in the level of neural activity. There are two basic paradigms that have been applied in functional neuroimaging of tinnitus. Firstly, sound-evoked responses as well as steady state neural activity have been measured to compare tinnitus patients to healthy controls. Secondly, paradigms that involve modulation of tinnitus by a controlled stimulus allow for a within-subject comparison that identifies neural activity that may be correlated to the tinnitus percept. Even though there are many differences across studies, the general trend emerging from the neuroimaging studies, is that tinnitus in humans may correspond to enhanced neural activity across several centers of the central auditory system. Also, neural activity in non-auditory areas including the frontal areas, the limbic system and the cerebellum seems associated with the perception of tinnitus. These results indicate that in addition to the auditory system, non auditory systems may represent a neural correlate of tinnitus. Although the currently published neuroimaging studies typically show a correspondence between tinnitus and enhanced neural activity, it will be important to perform future studies on subject groups that are closely matched for characteristics such as age, gender and hearing loss in order to rule out the contribution of these factors to the abnormalities specifically ascribed to tinnitus. PMID- 19545618 TI - Mechanisms of p,p'-DDE-induced apoptosis in human peripheral blood mononuclear cells. AB - p,p'-Dichlorodiphenyldichloroethylene (DDE), the most stable metabolite of organochlorine insecticide p,p'-dichlorodiphenyltrichloroethane (DDT), has been detected in human populations living in malaria-endemic areas of Mexico where this insecticide was used. DDE induces apoptosis in peripheral blood mononuclear cells (PMBC); however, the molecular mechanism of cell death induced by this compound is poorly understood. In the present study, PBMC isolated from healthy individuals (not exposed to DDE) were incubated in the presence of increasing concentrations of p,p'-DDE (0-80 microg/ml) over time. When PBMC were treated with low p,p'-DDE concentration (10 microg/ml) an antioxidant response and biomarkers of inflammation were induced, indicating a pro-inflammatory state. Moreover, when PBMC were treated with high p,p'-DDE concentration (80 microg/ml) several apoptotic biochemical events were triggered, such as activation of caspase-8, Bid, caspase-9 and caspase-3, as well as degradation of PARP and ubiquitination. The results described in this study show a possible inflammatory condition and the involvement of both extrinsic and intrinsic pathways in the induction of apoptosis in DDE-treated PBMC. PMID- 19545619 TI - Nitric oxide bioactivity of traditional Chinese medicines used for cardiovascular indications. AB - Traditional Chinese medicine (TCM) has been used for centuries to treat and prevent certain ailments and diseases. Although TCM has served as mainstream medical care throughout Asia for many generations, it is considered an alternative medical system in much of the Western world. Because many TCMs are used primarily for cardiovascular indications characterized by a nitric oxide (NO) insufficiency, we hypothesized that some, if not all, of these TCMs have a robust NO bioactivity that may act to restore NO homeostasis. We tested a group of convenience samples of TCMs obtained in the United States for endogenous nitrite, nitrate, nitroso, and nitrite reductase activity as well as their ability to relax isolated aortic rings. The results from this study reveal that all of the TCMs tested reveal NO bioactivity through their inherent nitrite and nitrate content and their ability to reduce nitrite to NO. Many of the TCM extracts contain a nitrite reductase activity greater by 1000 times that of biological tissues. Repletion of biological nitrite and nitrate by these extracts and providing a natural system for NO generation in both endothelium-dependent and -independent mechanisms may account for some of the therapeutic effects of TCMs. PMID- 19545621 TI - Inhibition of mitochondria-dependent apoptosis by 635-nm irradiation in sodium nitroprusside-treated SH-SY5Y cells. AB - Nitric oxide (NO) is a major factor contributing to the loss of neurons in ischemic stroke, demyelinating diseases, and other neurodegenerative disorders. NO not only functions as a direct neurotoxin, but also combines with superoxide (O(2)(-)) by a diffusion-controlled reaction to form peroxynitrite (ONOO(-)), a species that contributes to oxidative signaling and cellular apoptosis. However, the mechanism by which ONOO(-) induces apoptosis remains unclear, although subsequent formation of reactive oxygen species (ROS) has been suggested. The aim of this study was to further investigate the triggers of the apoptotic pathway using O(2)(-) scavenging with light irradiation to block ONOO(-) production. Antiapoptotic effects of light irradiation in sodium nitroprusside (SNP)-treated SH-SY5Y cells were assayed by reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide, DNA fragmentation, flow cytometry, Western blot, and caspase activity assays. In addition, NO, total ROS, O(2)(-), and ONOO(-) levels were measured to observe changes in NO and its possible involvement in radical induction. Cell survival was reduced to approximately 40% of control levels by SNP treatment, and this reduction was increased to 60% by low-level light irradiation. Apoptotic cells were observed in the SNP-treated group, but the frequency of these was reduced in the irradiation group. NO, O(2)(-), total ROS, and ONOO(-) levels were increased after SNP treatment, but O(2)(-), total ROS, and ONOO(-) levels were decreased after irradiation, despite the high NO concentration induced by SNP treatment. Cytochrome c was released from mitochondria of SNP-treated SH-SY5Y cells, but not of irradiated cells, resulting in a decrease in caspase-3 and -9 activity in SNP-treated cells. Finally, these results show that 635-nm irradiation, by promoting the scavenging of O(2)(-), protected against neuronal death through blocking the mitochondrial apoptotic pathway induced by ONOO(-) synthesis. PMID- 19545620 TI - Sod2 knockdown in the musculature has whole-organism consequences in Drosophila. AB - Oxidative damage to cell macromolecules by reactive oxygen species is associated with numerous diseases and aging. In Drosophila, RNAi-mediated silencing of the mitochondrial antioxidant manganese superoxide dismutase (SOD2) throughout the body dramatically reduces life span, accelerates senescence of locomotor function, and enhances sensitivity to applied oxidative stress. Here, we show that Sod2 knockdown in the musculature alone is sufficient to cause the shortened life span and accelerated locomotor declines observed with knockdown of Sod2 throughout the body, indicating that Sod2 deficiency in muscle is central to these phenotypes. Knockdown of Sod2 in the muscle also increased caspase activity (a marker for apoptosis) and caused a mitochondrial pathology characterized by swollen mitochondria, decreased mitochondrial content, and reduced ATP levels. These findings indicate that Sod2 plays a crucial role in the musculature in Drosophila and that the consequences of SOD2 loss in this tissue extend to the viability of the organism as a whole. PMID- 19545622 TI - CaMKII knockdown attenuates H2O2-induced phosphorylation of ERK1/2, PKB/Akt, and IGF-1R in vascular smooth muscle cells. AB - We have shown earlier a requirement for Ca(2+) and calmodulin (CaM) in the H(2)O(2)-induced activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) and protein kinase B (PKB), key mediators of growth-promoting, proliferative, and hypertrophic responses in vascular smooth muscle cells (VSMC). Because the effect of CaM is mediated through CaM-dependent protein kinase II (CaMKII), we have investigated here the potential role of CaMKII in H(2)O(2) induced ERK1/2 and PKB phosphorylation by using pharmacological inhibitors of CaM and CaMKII, a CaMKII inhibitor peptide, and siRNA knockdown strategies for CaMKII alpha. Calmidazolium and W-7, antagonists of CaM, as well as KN-93, a specific inhibitor of CaMKII, attenuated H(2)O(2)-induced responses of ERK1/2 and PKB phosphorylation in a dose-dependent fashion. Similar to H(2)O(2), calmidazolium and KN-93 also exhibited an inhibitory effect on glucose/glucose oxidase-induced phosphorylation of ERK1/2 and PKB in these cells. Transfection of VSMC with CaMKII autoinhibitory peptide corresponding to the autoinhibitory domain (aa 281 309) of CaMKII and with siRNA of CaMKII alpha attenuated the H(2)O(2)-induced phosphorylation of ERK1/2 and PKB. In addition, calmidazolium and KN-93 blocked H(2)O(2)-induced Pyk2 and insulin-like growth factor-1 receptor (IGF-1R) phosphorylation. Moreover, treatment of VSMC with CaMKII alpha siRNA abolished the H(2)O(2)-induced IGF-1R phosphorylation. H(2)O(2) treatment also induced Thr(286) phosphorylation of CaMKII, which was inhibited by both calmidazolium and KN-93. These results demonstrate that CaMKII plays a critical upstream role in mediating the effects of H(2)O(2) on ERK1/2, PKB, and IGF-1R phosphorylation. PMID- 19545623 TI - Myoglobin causes oxidative stress, increase of NO production and dysfunction of kidney's mitochondria. AB - Rhabdomyolysis or crush syndrome is a pathology caused by muscle injury resulting in acute renal failure. The latest data give strong evidence that this syndrome caused by accumulation of muscle breakdown products in the blood stream is associated with oxidative stress with primary role of mitochondria. In order to evaluate the significance of oxidative stress under rhabdomyolysis we explored the direct effect of myoglobin on renal tubules and isolated kidney mitochondria while measuring mitochondrial respiratory control, production of reactive oxygen and nitrogen species and lipid peroxidation. In parallel, we evaluated mitochondrial damage under myoglobinurea in vivo. An increase of lipid peroxidation products in kidney mitochondria and release of cytochrome c was detected on the first day of myoglobinuria. In mitochondria incubated with myoglobin we detected respiratory control drop, uncoupling of oxidative phosphorylation, an increase of lipid peroxidation products and stimulated NO synthesis. Mitochondrial pore inhibitor, cyclosporine A, mitochondria-targeted antioxidant (SkQ1) and deferoxamine (Fe-chelator and ferryl-myoglobin reducer) abrogated these events. Similar effects (oxidative stress and mitochondrial dysfunction) were revealed when myoglobin was added to isolated renal tubules. Thus, rhabdomyolysis can be considered as oxidative stress-mediated pathology with mitochondria to be the primary target and possibly the source of reactive oxygen and nitrogen species. We speculate that rhabdomyolysis-induced kidney damage involves direct interaction of myoglobin with mitochondria possibly resulting in iron ions release from myoglobin's heme, which promotes the peroxidation of mitochondrial membranes. Usage of mitochondrial permeability transition blockers, Fe-chelators or mitochondria-targeted antioxidants, may bring salvage from this pathology. PMID- 19545624 TI - Intra-articular drug delivery systems for the treatment of rheumatic diseases: a review of the factors influencing their performance. AB - Osteoarthritis and rheumatoid arthritis are rheumatic diseases for which a curative treatment does not currently exist. Their management is directed towards pain relief achieved with different classes of drugs among which non-steroidal and steroidal anti-inflammatory substances are the most frequently used agents. Nevertheless, the oral or systemic administration of such drugs is hindered by numerous side effects, which could be overcome by their intra-articular (i-a.) administration as dosage forms capable of gradually releasing the active substance. The present review article summarises the research done in the field of drug delivery systems for i-a. injection vs. current management of osteoarthritis or rheumatoid arthritis. Aspects such as the influence of size, shape, polymer matrix or targeted drug on the i-a. retention time, phagocytosis and biological activity will be discussed. Finally, we will comment on the need for adapted delivery systems for the novel and very potent anti-inflammatory drugs, such as inhibitors of the p38 mitogen-activated protein kinase or the IL 1beta conversion enzyme, which to date cannot be properly used due to the severe side effects associated with their systemic administration. PMID- 19545625 TI - Non-invasive in vivo evaluation of in situ forming PLGA implants by benchtop magnetic resonance imaging (BT-MRI) and EPR spectroscopy. AB - In the present study, we used benchtop magnetic resonance imaging (BT-MRI) for non-invasive and continuous in vivo studies of in situ forming poly(lactide-co glycolide) (PLGA) implants without the use of contrast agents. Polyethylene glycol (PEG) 400 was used as an alternative solvent to the clinically used NMP. In addition to BT-MRI, we applied electron paramagnetic resonance (EPR) spectroscopy to characterize implant formation and drug delivery processes in vitro and in vivo. We were able to follow key processes of implant formation by EPR and MRI. Because EPR spectra are sensitive to polarity and mobility, we were able to follow the kinetics of the solvent/non-solvent exchange and the PLGA precipitation. Due to the high water affinity of PEG 400, we observed a transient accumulation of water in the implant neighbourhood. Furthermore, we detected the encapsulation by BT-MRI of the implant as a response of the biological system to the polymer, followed by degradation over a period of two months. We could show that MRI in general has the potential to get new insights in the in vivo fate of in situ forming implants. The study also clearly shows that BT-MRI is a new viable and much less expensive alternative for superconducting MRI machines to monitor drug delivery processes in vivo in small mammals. PMID- 19545626 TI - Oleoyl-estrone affects lipid metabolism in adrenalectomized rats treated with corticosterone through modulation of SREBP1c expression. AB - Oleoyl-estrone (OE) elicits a decrease in body fat, which is blocked by glucocorticoids. In order to analyze this counterregulatory effect, we studied the effects of oral OE on adrenalectomized female rats simultaneously receiving corticosterone (subcutaneous pellets). Circulating corticosteroids, liver glycogen, lipids and the expressions in whole liver, soleus muscle, interscapular brown adipose tissue (BAT), and the inguinal and periovaric white adipose tissue (WAT) of genes controlling lipid metabolism were analyzed. Corticosterone reversed OE lipid mobilization, storing fat in liver and subcutaneous WAT. This was not simply the predominance of corticosteroid enhancement of lipogenesis against OE inhibition, but a synergy to enhance lipogenesis. Periovaric WAT showed a different effect, with corticosterone inhibiting OE arrest of lipogenic gene expressions. The data presented suggests that interaction of OE and glucocorticoids (and the metabolic response) depends on the organ or WAT site; there was a direct relationship on the direction and extent of change of SREBP1c expression with those of important energy and lipid handling genes. Our results confirm that corticosterone blocks - and even reverses - OE effects on body lipids in a dose-dependent way, a process mediated, at least in part, by modulation of SREBP1c expression. PMID- 19545627 TI - Towards Coleoptera-specific high-throughput screening systems for compounds with ecdysone activity: development of EcR reporter assays using weevil (Anthonomus grandis)-derived cell lines and in silico analysis of ligand binding to A. grandis EcR ligand-binding pocket. AB - Molting in insects is regulated by ecdysteroids and juvenile hormones. Several synthetic non-steroidal ecdysone agonists are on the market as insecticides. These ecdysone agonists are dibenzoylhydrazine (DBH) analogue compounds that manifest their toxicity via interaction with the ecdysone receptor (EcR). Of the four commercial available ecdysone agonists, three (tebufenozide, methoxyfenozide and chromafenozide) are highly lepidopteran specific, one (halofenozide) is used to control coleopteran and lepidopteran insects in turf and ornamentals. However, compared to the very high binding affinity of these DBH analogues to lepidopteran EcRs, halofenozide has a low binding affinity for coleopteran EcRs. For the discovery of ecdysone agonists that target non-lepidopteran insect groups, efficient screening systems that are based on the activation of the EcR are needed. We report here the development and evaluation of two coleopteran-specific reporter-based screening systems to discover and evaluate ecdysone agonists. The screening systems are based on the cell lines BRL-AG-3A and BRL-AG-3C that are derived from the weevil Anthonomus grandis, which can be efficiently transduced with an EcR reporter cassette for evaluation of induction of reporter activity by ecdysone agonists. We also cloned the almost full length coding sequence of EcR expressed in the cell line BRL-AG-3C and used it to make an initial in silico 3D model of its ligand-binding pocket docked with ponasterone A and tebufenozide. PMID- 19545628 TI - In vivo role for CREB signaling in the noradrenergic differentiation of sympathetic neurons. AB - Signaling pathways involving cAMP and CREB have been implicated in several aspects of sympathetic neuron differentiation. Here, we used in vivo loss-of function approaches in both mouse and chick embryos to characterize the physiological role of cAMP/CREB. Whereas sympathetic neuron development proceeds normally in CREB-deficient mouse embryos, a decrease in noradrenergic differentiation (TH, DBH) was observed in chick sympathetic ganglia in response to ACREB, a dominant-negative CREB variant which interferes with the function of all CREB family members. In contrast, expression of the generic neuronal marker SCG10 was not affected by ACREB. As the decrease in noradrenergic gene expression is compensated at later stages of development and TH expression in differentiated neurons is not CREB-dependent, a transient role for CREB is proposed, accelerating noradrenergic but not generic neuronal differentiation of sympathetic neurons. PMID- 19545631 TI - WITHDRAWN: High level expression and characterization of Aspergillus niger beta mannanase in Methylotrophic yeast. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19545630 TI - Intranasal administration of proteoliposome-derived cochleates from Vibrio cholerae O1 induce mucosal and systemic immune responses in mice. AB - Conservative estimates place the death toll from cholera at more than 100,000 persons each year. A particulate mucosal vaccine strategy combining antigens and immune stimulator molecules from Vibrio cholerae to overcome this problem is described. Proteoliposomes extracted from V. cholerae O1 were transformed into cochleates (AFCo2, Adjuvant Finlay cochleate 2) through a calcium inducible rotary dialysis method. Light microscopy was carried out and tubules of 16.25+/ 4.57 microm in length were observed. Western blots were performed to verify the immunochemical properties of the main AFCo2 incorporated antigens, revealing full recognition of the outer membrane protein U (OmpU), lipopolysaccharide (LPS), and mannose-sensitive hemagglutinin (MSHA) antigens. AFCo2 were administered by the intranasal route using a two or three dose schedule and the immune response against V. cholerae antigens was assessed. Three AFCo2 doses were required to induce significant (p<0.05), antigen specific IgA in saliva (1.34+/-0.135) and feces (0.60+/-0.089). While, two or three doses of AFCo2 or proteoliposomes induce similar specific IgG and vibriocidal activity responses in sera. These results show for the first time that AFCo2 can be obtained from V. cholerae O1 proteoliposomes and have the potential to protect against the pathogen when administered intranasally. PMID- 19545629 TI - Copy number variants (CNVs) in primate species using array-based comparative genomic hybridization. AB - A substantial amount of genomic variation is now known to exist in humans and other primate species. Single nucleotide polymorphisms (SNPs) are thought to represent the vast majority of genomic differences among individuals in a given primate species and comprise about 0.1% of the genomes of two humans. However, recent studies have now shown that structural variation msay account for as much as 0.7% of the genomic differences in humans, of which copy number variants (CNVs) are the largest component. CNVs are segments of DNA that can range in size from hundreds of bases to millions of base pairs in length and have different number of copies between individuals. Recent technological advancements in array technologies led to genome-wide identification of CNVs and consequently revealed thousands of variable loci in humans, comprising as much as 12% of the human genome [A.J. Iafrate, L. Feuk, M.N. Rivera, M.L. Listewnik, P.K. Donahoe, Y. Qi, S.W. Scherer, C. Lee, Nat. Genet. 36 (2004) 949-951, [3]]. CNVs in humans have already been associated with susceptibility to certain complex diseases, dietary adaptation, and several neurological conditions. In addition, recent studies have shown that CNVs can be successfully implemented in population genetics research, providing important insights into human genetic variation. Nevertheless, the important role of CNVs in primate evolution and genetic diversity is still largely unknown. This article aims to outline the strengths and weaknesses of current comparative genomic hybridization array technologies that have been employed to detect CNV variation and the applications of these techniques to primate genetic research. PMID- 19545632 TI - Expression and purification of APRIL by auto-induction. AB - APRIL (A proliferation-inducing ligand) is a newly-identified member of the tumor necrosis factor family that induces pleiotropic biological responses, including immunological responses, IgA class switch and cell growth. It is associated with multiple diseases such as cancer and autoimmune diseases. High levels of APRIL mRNA can be detected in transformed cell lines and several malignant tumors; heparin sulfate proteoglycans (HSPG) are also involved in the APRIL tumor cell proliferation induction response. The interaction of APRIL and HSPG occurs through an N-terminal basic region on APRIL. We successfully expressed recombinant APRIL using an auto-induction system in Escherichia coli. By using in situ cleavage, we generated the mature form of APRIL, comprising its N-terminal basic region, which has full biological activity: receptor binding capability and proliferation induction activity. PMID- 19545633 TI - High yield and purification of recombinant human apolipoprotein E3 in Pichia pastoris. AB - Apolipoprotein E3 (ApoE3) is an important apolipoprotein in plasma and plays a critical role in lipid transport and cholesterol homeostasis. As the only natural source of this protein, human blood cannot provide large-scale ApoE3 for research and applications. Therefore, in our study, a Pichia pastoris expression system was first used to obtain a high-level expression of secreted, recombinant human ApoE3 (rhApoE3). The full-length sequence encoding ApoE3, gained by RT-PCR, was inserted into the pPICZalphaC vector and transformed into P. pastoris strain X33, and then the high expression transformants with zeocin resistance were obtained. The growth conditions of the transformant strains were optimized in 50ml conical tubes including pH and inducing time. After induction with methanol, the expression level of rhApoE3 was 120 mg/L in 80 L fermentor. RhApoE3 was purified more than 94% purity using SP Sepharose ion exchange chromatography and source 30RPC. A preliminary biochemical characterization of purified rhApoE3 was performed by analyzing the ability of inhibiting PDGF-induced proliferation of rat coronary artery smooth muscle cells (SMCs), and the results demonstrated that the function of purified rhApoE3 was similar to natural human ApoE3. PMID- 19545634 TI - High level expression and purification of active recombinant human interleukin-8 in Pichia pastoris. AB - Human interleukin-8 (hIL-8) is a member of interleukin family which functions as a chemotactic factor as well as an angiogenesis mediator. Previously, a study reported that hIL-8 could be purified from inclusion bodies using a prokaryotic expression system, however, the required re-naturation step limits the recovery of fully active protein. In this study, soluble recombinant hIL-8 was expressed as a secreted protein at high level in Pichia pastoris under the control of AOX1 (alcohol oxidase 1) promoter. A simple purification strategy was established to recover rhIL-8 from the fermentation supernatant. The process includes precipitation with 80% saturation ammonium sulfate and CM Sepharose ion exchange chromatography, yielding 30 mg/L purified rhIL-8 at over 95% purity. The obtained rhIL-8 displays high specific activity, stimulating the migration of mouse neutrophils at concentrations as low as 0.25 ng/mL. Our results demonstrate that P. pastoris expression system is an efficient tool for large-scale manufacture of active recombinant hIL-8 for various applications. PMID- 19545635 TI - Combining electron crystallography and X-ray crystallography to study the MlotiK1 cyclic nucleotide-regulated potassium channel. AB - We have recently reported the X-ray structure of the cyclic nucleotide-regulated potassium channel, MlotiK1. Here we describe the application of both electron and X-ray crystallography to obtain high quality crystals. We suggest that the combined application of these techniques provides a useful strategy for membrane protein structure determination. We also present negative stain projection and cryo-data projection maps. These maps provide new insights about the properties of the MlotiK1 channel. In particular, a comparison of a 9A cryo-data projection with calculated model maps strongly suggests that there is a very weak interaction between the pore and the S1-S4 domains of this 6 TM tetrameric cation channel and that the S1-S4 domains can adopt multiple orientations relative to the pore. PMID- 19545636 TI - The AAA(+) motor complex of subunits CobS and CobT of cobaltochelatase visualized by single particle electron microscopy. AB - Cobalamins belong to the tetrapyrrole family of prosthetic groups. The presence of a metal ion is a key feature of these compounds. In the oxygen-dependent (aerobic) cobalamin biosynthetic pathway, cobalt is inserted into a ring contracted tetrapyrrole called hydrogenobyrinic acid a,c-diamide (HBAD) by a cobaltochelatase that is constituted by three subunits, CobN, CobS and CobT, with molecular masses of 137, 37 and 71kDa, respectively. Based on the similarities with magnesium chelatase, cobaltochelatase has been suggested to belong to the AAA(+) superfamily of proteins. In this paper we present the cloning of the Brucella melitensis cobN, cobS and cobT, the purification of the encoded protein products, and a single-particle reconstruction of the macromolecular assembly formed between CobS and CobT from negatively stained electron microscopy images of the complex. The results show for the first time that subunits CobS and CobT form a chaperone-like complex, characteristic for the AAA(+) class of proteins. The molecules are arranged in a two-tiered ring structure with the six subunits in each ring organized as a trimer of dimers. The similarity between this structure and that of magnesium chelatase, as well as analysis of the amino acid sequences confirms the suggested evolutionary relationship between the two enzymes. PMID- 19545637 TI - Dual-axis target mapping and automated sequential acquisition of dual-axis EM tomographic data. AB - Dual-axis electron microscopic tomography minimizes the missing wedge-induced resolution loss by taking two complementary tilt data sets of the same target along two orthogonal axes. The potential of this powerful approach has been hampered by the practical challenges inherent in finding the original targets that are dramatically displaced due to non-eucentric specimen rotation. Not only is the manual search for the original targets time consuming and tedious but the added dose during manual searching is uncontrollable. We have developed a hierarchical alignment scheme that allows tomographic data to be collected from an arbitrary number of target sites in one grid orientation and then to find and collect orthogonal data sets with little or no user intervention. Inspired by the successful multi-scale mapping in Leginon, our alignment is performed in three levels to gradually pinpoint the original targets. At the lowest level the grid lattice is used to determine the rotation angle and translational shift resulting from specimen rotation via auto- and cross-correlative analysis of a pair of atlas maps constructed before and after specimen rotation. The target locations are further refined at the next level using a pair of smaller atlas maps. The final refinement of target positions is done by aligning the target contained image tiles. Given the batch processing nature of this hierarchical alignment, multiple targets are initially selected in a group and then sequentially acquired. Upon completion of the data collection on all the targets along the first axis and after specimen rotation, the hierarchical alignment is performed to relocate the original targets. The data collection is then resumed on these targets for the second axis. Therefore, only one specimen rotation is needed for collecting multiple dual-axis tomographic data sets. The experiment of acquiring 20S Proteasomes dual-axis tomographic data sets in vitreous ice at 86,000x CCD magnification on our FEI Tecnai Polara TF30 electron microscope has suggested that the developed scheme is very robust. The extra doses for finding and centering the original targets are almost negligible. This scheme has been integrated into UCSF Tomography software suite that can be downloaded at www.msg.ucsf.edu/tomography free for academic use. PMID- 19545638 TI - Genomic organization and expression analysis of Toll-like receptor 3 in grass carp (Ctenopharyngodon idella). AB - Toll-like receptor 3 (TLR3) participates in the innate immune response by recognizing viral pathogens. To investigate grass carp immune system responding to GCRV (grass carp reovirus) infection, the full-length cDNA sequence and genomic organization of grass carp TLR3 (CiTLR3) was identified and characterized. The full-length genome sequence of CiTLR3 is composed of 5668 nucleotides, including five exons and four introns. The full-length of CiTLR3 cDNA is 3681 bp in length and encodes a polypeptide of 904 amino acids with an estimated molecular mass of 102,765 Da and a predicted isoelectric point of 8.35. Analysis of the deduced amino acid sequence indicated that CiTLR3 has four main structural domains, including a signal peptide sequence, 14 LRR (leucine-rich repeat) motifs, a transmembrane region and a TIR (Toll/interleukin-1 receptor) domain. It is most similar to the crucian carp (Carassius auratus) TLR3 amino acid sequence with an identity of 99%. Quantitative RT-PCR analysis showed that CiTLR3 transcripts were significantly up-regulated starting at day 1 and continued through day 7 following GCRV infection (P < 0.05). These data implied that CiTLR3 is involved in antiviral defense, provide molecular and functional information for grass carp TLR3, and implicate their role in mediating immune protection against grass carp viral diseases. PMID- 19545639 TI - Methods of maintaining human embryonic stem cells continues to be worth studying. AB - This article gives a brief comment on the culture of human embryonic stem cells (hESCs) with the aim to maintain the potency of hESCs in well state and produce more homogenous cell clones. PMID- 19545640 TI - A double WAP domain (DWD)-containing protein with proteinase inhibitory activity in Chinese white shrimp, Fenneropenaeus chinensis. AB - The whey major component, whey acidic protein (WAP), has one or more WAP domains characterized by a four-disulfide core (4-DSC) structure. These kinds of proteins are involved in multiple functions, including proteinase inhibitor activity, antimicrobial activity, ATPase inhibitor activity, and regulatory function in cell proliferation. Recent research indicates that WAP domain-containing proteins play an important role in the innate immunity of crustaceans. In this study, a novel double WAP domain (DWD)-containing protein named Fc-DWD was found for the first time in Chinese white shrimp, Fenneropenaeus chinensis. The open reading frame of Fc-DWD encodes a protein of 117 amino acids, including a signal peptide of 16 amino acids and two WAP domains. The predicted molecular mass of the mature protein is 12.78kDa with an estimated pI of 8.49. The first WAP domain, named WAP 1, composed of 49 amino acids locates in the amino-terminal of Fc-DWD, and the second WAP domain, named WAP 2, composed of 45 amino acids locates in the carboxy terminal. Fc-DWD mRNA was upregulated in hemocytes, hepatopancreas, gills, and stomach of bacteria- and virus-challenged shrimp. Results of the binding assay showed that rFc-DWD could bind to both Gram-negative bacteria and Gram-positive bacteria. rWAP 1 could only bind to Gram-positive bacteria, but rWAP 2 could bind to both Gram-negative and positive bacteria. Moreover, rFc-DWD exhibited proteinase inhibitory activity against the secretory proteinase(s) from Bacillussubtilis and Pseudomonas aeruginosa. All of these findings suggest that Fc-DWD may play an important role in enabling the host defense to execute its proteinase inhibitory activity against pathogens. PMID- 19545641 TI - Molecular cloning, mRNA expression and enzymatic characterization of cathepsin F from olive flounder (Paralichthys olivaceus). AB - Cathepsin F is a recently described papain-like cysteine protease of unknown function, and unique among cathepsins due to an elongated N-terminal pro-region, which contains a cystatin domain. In the present study, the cDNA of olive flounder (Paralichthys olivaceus) cathepsin F (PoCtF) was cloned by the combination of homology molecular cloning and rapid amplification of cDNA ends (RACE) approaches. The PoCtF gene was determined to consist of the 1844 bp nucleotide sequence which encodes for a 475-amino acid polypeptide. The results of RT-PCR analysis revealed ubiquitous expression throughout the entirety of healthy flounder tissues; however the PoCtF expressions increased significantly in gill at 3h post-injection with lipopolysaccharide (LPS). Also, immunostaining using anti-PoCtF antibody was strongest on the epidermal mucus in the fin. The cDNA encoding mature enzyme of PoCtF was expressed in Escherichia coli using the pGEX-4T-1 expression vector system. Its activity was quantified by cleaving the synthetic peptide Z-Phe-Arg-AMC, a substrate commonly used for functional characterization of cysteine proteinases, and the optimal pH for the protease activity was 7.5. The findings of the present study suggest that PoCtF has a higher optimum pH than mammalian cathepsin F, and PoCtF is an interesting target for future investigations of the role of cathepsin F in the epidermal mucus and fish innate immune system. PMID- 19545643 TI - Mitochondria matter: new concepts of dynamics and roles in pathophysiology. PMID- 19545644 TI - Approaches to seizure risk assessment in preclinical drug discovery. AB - Assessment of seizure risk traditionally occurs late in the drug discovery process using low-throughput, resource intensive in vivo assays. Such approaches do not allow sufficient time to mitigate risk by influencing chemical design. Early identification using cheaper, higher throughput assays with lower animal and compound requirements would be preferable. Here we review the current techniques available to assess this issue and describe how they may be combined in a rational step-wise cascade allowing more effective assessment of seizure risk. PMID- 19545646 TI - Induction of SCEs in CHL cells by dichlorobiphenyl derivative water pollutants, 2 phenylbenzotriazole (PBTA) congeners and river water concentrates. AB - We recently identified dichlorobiphenyl (DCB) derivatives and 2 phenylbenzotriazole (PBTA) congeners as major mutagenic constituents of the waters of the Waka River and the Yodo River system in Japan, respectively. In this study we examined sister chromatid exchange (SCE) induction by two dichlorobiphenyl derivatives, 3,3'-dichlorobenzidine (DCB, 4,4'-diamino-3,3' dichlorobiphenyl) and 4,4'-diamino-3,3'-dichloro-5-nitrobiphenyl (5-nitro-DCB); three PBTA congeners, 2-[2-(acetylamino)-4-[bis(2-methoxyethyl)amino]-5 methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA-1), 2-[2 (acetylamino)-4-[N-(2-cyanoethyl)ethylamino]-5-methoxyphenyl]-5-amino-7-bromo-4 chloro-2H-benzotriazole (PBTA-2), and 2-[2-(acetylamino)amino]-4-[bis(2 hydroxyethyl)amino]-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA-6); and water concentrates from the Waka River in Chinese hamster lung (CHL) cells. Concentration-dependent induction of SCE was found for all DCBs and PBTAs examined in the presence of S9 mix, and statistically significant increases of SCEs were detected at 2 microg per ml of medium or higher concentrations. SCE induction of MeIQx was examined to compare genotoxic activities of these water pollutants. According to the results, a ranking of the SCE-inducing potency of these compounds is the following: 5-nitro-DCB approximately MeIQx>PBTA6>PBTA-1 approximately PBTA-2>DCB. Water samples collected at a site at the Waka River showed concentration-related increases in SCEs at 6.25-18.75 ml-equivalent of river water per ml of medium with S9 mix. The concentrations of 5-nitro-DCB and DCB in the river water samples were from 2.5 to 19.4 ng/l and from 4100 to 18,900 ng/l, respectively. However, these chemicals showed only small contribution to SCE induction by the Waka River water. PMID- 19545645 TI - Chromosomal aberrations in a fish, Channa punctata after in vivo exposure to three heavy metals. AB - The studies were designed to assess the extent of chromosomal aberrations (CA) under the exposure of three common heavy metalic compounds, viz. mercuric chloride, arsenic trioxide and copper sulphate pentahydrate, in vivo using fish, Channa punctata (2n=32), as a test model. Prior acclimatized fishes were divided into five groups. Group I and II served as negative and positive control, respectively. An intramuscular injection of Mitomycin-C (@ 1mg/kg body wt.) was administered to group II only. Fishes of groups III, IV and V were subjected to sublethal concentrations (10% of 96h LC(50)), of HgCl(2) (0.081mg/L), As(2)O(3) (6.936mg/L) and CuSO(4)x5H(2)O (0.407mg/L). Fishes of all the groups were exposed uninterrupted for 24, 48, 72, 96 and 168h. Observations of kidney cells of exposed fishes revealed chromatid and chromosome breaks, chromatid and chromosome gaps along with ring and di-centric chromosomes. A significant increase over negative control in the frequency of chromosomal aberrations (CA) was observed in fish exposed to Mitomycin-C, Hg(II), As(III) and Cu(II). As the average + or - SE total number of CA, average number of CA per metaphase and %incidence of aberrant cells in Hg(II) was 104.40 + or - 8.189, 0.347 + or - 0.027 and 10.220 + or - 0.842, respectively; in As(III) 109.20 + or - 8.309, 0.363 + or - 0.027 and 10.820 + or - 2.347, respectively and in Cu(II) 89.00 + or - 19.066, 0.297 + or - 0.028 and 8.900 + or - 0.853, respectively. Hence, it reveals that the order of induction of frequency of CA was Cu35 years. A historical perspective and the basic features of DNA polymerase proofreading are described here, but the goal of this review is to present recent advances in the elucidation of the proofreading pathway and to describe roles of DNA polymerase proofreading beyond mismatch correction that are also important for maintaining genome stability. PMID- 19545650 TI - Research on an Mg-Zn alloy as a degradable biomaterial. AB - In this study a binary Mg-Zn magnesium alloy was researched as a degradable biomedical material. An Mg-Zn alloy fabricated with high-purity raw materials and using a clean melting process had very low levels of impurities. After solid solution treatment and hot working the grain size of the Mg-Zn alloy was finer and a uniform single phase was gained. The mechanical properties of this Mg-Zn alloy were suitable for implant applications, i.e. the tensile strength and elongation achieved were approximately 279.5MPa and 18.8%, respectively. The results of in vitro degradation experiments including electrochemical measurements and immersion tests revealed that the zinc could elevate the corrosion potential of Mg in simulated body fluid (SBF) and reduce the degradation rate. The corrosion products on the surface of Mg-Zn were hydroxyapatite (HA) and other Mg/Ca phosphates in SBF. In addition, the influence caused by in vitro degradation on mechanical properties was studied, and the results showed that the bending strength of Mg-Zn alloy dropped sharply in the earlier stage of degradation, while smoothly during the later period. The in vitro cytotoxicity of Mg-Zn was examined. The result 0-1 grade revealed that the Mg-Zn alloy was harmless to L-929 cells. For in vivo experiments, Mg-Zn rods were implanted into the femoral shaft of rabbits. The radiographs illustrated that the magnesium alloy could be gradually absorbed in vivo at about 2.32mm/yr degradation rate obtained by weight loss method. Hematoxylin and eosin (HE) stained section around Mg-Zn rods suggested that there were newly formed bone surrounding the implant. HE stained tissue (containing heart, liver, kidney and spleen tissues) and the biochemical measurements, including serum magnesium, serum creatinine (CREA), blood urea nitrogen (BUN), glutamic-pyruvic transaminase (GPT) and creatine kinase (CK) proved that the in vivo degradation of Mg-Zn did not harm the important organs. Moreover, no adverse effects of hydrogen generated by degradation had been observed and also no negative effects caused by the release of zinc were detected. These results suggested that the novel Mg-Zn binary alloy had good biocompatibility in vivo. PMID- 19545651 TI - Familial Sotos syndrome caused by a novel missense mutation, C2175S, in NSD1 and associated with normal intelligence, insulin dependent diabetes, bronchial asthma, and lipedema. AB - We report a familial Sotos syndrome in two children, boy and girl, aged 17 and 8 years, and in their 44 year old mother, who displayed normal intelligence at adult age, but suffered from insulin dependent diabetes mellitus, bronchial asthma, and severe lipedema. The underlying missense mutation, C2175S, occurred in a conserved segment of the NSD1 gene. Our findings confirm that familial cases of SS are more likely to carry missense mutations. This case report may prove useful to avoid underestimation of the recurrence rate of SS, and to demonstrate that the developmental delay may normalize, enabling an independent life and having an own family. PMID- 19545652 TI - Age-dependent systemic antibody responses and immunisation-associated changes in mice orally and nasally immunised with Lactococcus lactis expressing a malaria parasite protein. AB - Gram positive food-grade bacteria such as lactococci have significant advantages over attenuated pathogens as vaccine delivery vehicles because of their inherently greater safety. Plasmodium falciparum merozoite surface antigen 2 (MSA2) was expressed in recombinant Lactococcus lactis both intracellularly and covalently anchored to the peptidoglycan of the cell wall (MSA2cP). Balb/c mice of different ages were immunised with the MSA2cP expressing L. lactis in a combined oral and nasal immunisation procedure. Serum IgG antibody responses to MSA2 were higher in young adult Balb/c mice compared to old mice and neonates. The elicited serum IgG antibodies reacted with native MSA2 on the surface of P. falciparum merozoites in an immunofluorescence assay. The serum IgG antibody isotypes in young adult mice were mainly IgG1, IgG2a and IgG2b, while IgG3 tended to be higher in old mice. IgA antibodies to MSA2 were also produced in young mice. Enlarged mesenteric lymph nodes, and more prominent lymphoid tissue in the lamina propria of the ileum and lymphoid follicles in the spleen, were observed in mice fed L. lactis. These findings are relevant for developing L. lactis as a vector to deliver vaccines in human populations. PMID- 19545653 TI - Forty years of publication of transplantation proceedings--the second decade: the cyclosporine revolution. AB - With surgical tools in place, increased knowledge concerning immunogenetics and alloimmunity as well as improved management of immunocompromised patients, the foundations were lain for the rapid development of the transplantation enterprise. In contrast to pre-transplant conditioning by thoracic duct drainage or total lymphoid irradiation, which were too cumbersome for routine execution among the burgeoning recipient pool, cyclosporine was a facile method to produce immunosuppression de novo after transplantation. On the one hand, clinical data confirmed the potency of cyclosporine in a variety of clinical settings. On the other hand, a pleiotropic array of side effects, particularly nephrotoxicity, beclouded the regimens, even when used in reduced doses in combination with azathioprine or together with rigorous individualization of therapy by concentration control. The advent of cyclosporine condemned conditioning by pretransplant blood transfusions and donor-recipient HLA matching to therapeutic obsolescence. However, cross-matching achieved greater significance particularly due to the development of flow cytometry methods to detect modest amounts of anti donor antibody. Adjunctive treatments with polyclonal preparations or monoclonal antibodies were developed to provide an additional layer of security during the critical induction phase of immunosuppression and for treatment of rejection episodes refractory to high dose steroid therapy. Active immunologic investigation was stimulated by antibodies that discriminated CD4+ versus CD8+ T cells, leading to dissection of their numbers of precursors or mature elements as well as their distinct activities. The search for methods to induce, maintain and detect the state of transplantation tolerance continued. The encouraging results in clinical transplantation raised a variety of ethical concerns related to public attitudes; to retrieval, distribution, and allocation of the limited supply of deceased donor organs; the increased utilization of living persons; the opportunities for commerce; the quasi-righteous requests for organ gifts by unrelated individuals and the scant financial resources for long-term treatment with costly immunosuppression. Transplantation had now achieved its rightful place in the clinical armamentarium. PMID- 19545654 TI - Clinical impact of polymorphisms of transport proteins and enzymes involved in the metabolism of immunosuppressive drugs. AB - Individualization of immunosuppressive therapy after solid organ transplantation is a goal that has been pursued for a long time. Nevertheless, in clinical practice, we are still stratifying patients in subgroups in which risk is assessed using demographic information and population analysis. Then, a combination of immunosuppressive drugs is chosen and doses are individualized to compensate for intra- and interindividual variabilities in drug pharmacokinetics, to obtain similar plasma/blood concentrations that are believed to be therapeutic, again based on data derived from population analysis. One step further in this strategy is to recognize, before initiation of immunotherapy, those patients at higher risk to be either under- or overexposed to currently used immunosuppressive drugs. Several studies have been undertaken to correlate single nucleotide polymorphisms in genes encoding transport proteins and metabolizing enzymes involved in the disposition of immunosuppressive drugs. Overall, the results from these studies have been mixed. The causes of these sometimes conflicting results include methodologic, genetic, or nongenetic factors. The degree of linkage disequilibrium, the measure of nonrandom associations between polymorphisms at different loci, not necessarily on the same chromosome, is perhaps the main genetic factor. The influence of the environment, physiology (such as kidney and liver functions), disease state, use of multidrug regimens, and inherent drug-to-drug interactions are present nongenetic factors. Moreover, it is also important to increase our knowledge of the genetic factors involved in the variabilities observed in drug responses of pharmacodynamics. True individualized therapy, with the ability to improve health outcomes of each transplant recipient, will depend on our knowledge of the genetic factors involved in immunological response and drug pharmacokinetics and pharmacodynamics. PMID- 19545655 TI - Culture change initiatives in the procurement of organs in the United kingdom. AB - In 2004, the United Kingdom began for the first time a systematic, countrywide initiative to increase organ donations. The approach, based on international research and with consultation from the United States, entailed encouraging the early identification and referral of potential donors, training critical care physicians in breaking bad news and in collaborating with donor coordinators, training coordinators in donation conversation skills, and getting them in to speak with families. The strategy represented a large culture change in the procurement of organs in the United Kingdom and has met resistance. However, the change has started to take hold and, despite a dramatic and unexpected 21% reduction in potential brain stem death donors over a 4-year period, is showing positive results. Herein we have described the historical model for procuring organs in the United Kingdom, the development of the new strategy, its components, and the results of its implementation, in terms of earlier referrals, the identification of non-heart-beating donors, the collaboration between physicians and coordinators in speaking with families and in converted donors. PMID- 19545656 TI - Emergency department organ-donation card pilot study. AB - OBJECTIVE: To determine public attitudes about an emergency department (ED)-based organ-donor enrollment program. METHOD: A total of 211 non-acutely ill patients treated in the ED of a 300-bed urban university hospital between November 1, 2004, and January 2, 2005, were asked to complete a survey. Surveys were obtained over six 4-hour daytime intervals. RESULTS: Of the 211 patients, 199 completed the survey and 12 refused. One hundred sixty-four patients (82%; 95% confidence interval [CI], 77%-88%) were interested in organ-donation information via the ED; 173 (87%; 95% CI, 82%-92%) approved of the ED providing organ-donation cards; and 82 (41%; 95% CI, 34%-48%) were current organ-donor cardholders. Of 117 patients who were not current organ-donor cardholders, 65 (56%; 95% CI, 46%-66%) stated that they would be willing to enroll in an ED-based organ-donor card program. Fifty-seven patients (49%, 95% CI, 39%-58%) were African American, and 35 of these (61%; 95% CI, 48%-74%) said they would obtain an organ-donor card via the ED. Overall, 137 of 199 patients (69%; 95% CI, 62%-75%) preferred ED organ donation information in written form, and the remainder preferred a personal interview or lecture. CONCLUSIONS: An ED organ-donation enrollment program would be acceptable to patients and effective in capturing groups who currently do not have organ-donor cards. PMID- 19545657 TI - Contrast-enhanced transcranial color sonography in the diagnosis of cerebral circulatory arrest. AB - OBJECTIVE: We sought to determine the utility of constrat-enhanced transcranial color sonography (TCCS) in the diagnosis of cerebral circulatory arrest in cases of difficult acoustic window. MATERIALS AND METHODS: From January 2007 to July 2008, we prospectively studied 50 patients who fulfilled clinical criteria of brain death. In all cases, we performed TCCS aiming to insonate both middle cerebral arteries (MCA) and the basilar artery (BA). In those case in which insonation of any vessel was not possible, we repeated the exploration after injecting a 2.5-mL bolus of sulphurhexafluoridedispersion. Afterward, we compared the rate of insonation of the vessels and the number of conclusive studies. RESULTS: The mean patient age was 53.2 +/- 15.9 years. Thirty-two were men (64%). The most frequent neurologic injury was hemorrhagic stroke and traumatic brain injury. Contrast-enhanced TCCS resulted in an increased rate of insonation in both MCA and in BA, and in the number of conclusive studies. CONCLUSIONS: Contrast-enhanced TCCS increased the number of conclusive studies with cerebral circulatory arrest, which minimized the importance of a previous study in cases with a poor acoustic window. PMID- 19545658 TI - Knowledge and attitudes toward brain stem death among university undergraduates. AB - BACKGROUND: Withdrawal of life support and organ procurement for transplantation are the main implications of a diagnosis of brain stem death (BSD). Various factors may impact this important decision-making process. The present study sought to investigate the knowledge and attitudes about BSD among university undergraduates as a "well-informed" subgroup of our local population. METHODS: A cross-sectional questionnaire survey was administered to a sample of nonmedical university undergraduate students in Hong Kong. RESULTS: The subjects' overall knowledge of BSD was unsatisfactory. Only 24% of subjects knew that BSD was the equivalent of legal death in Hong Kong. Among subjects who agreed to withdraw life support treatment from themselves upon the diagnosis of BSD, 30% and 24% refused to do so for their family members or a stranger, respectively. Subjects who agreed to withdraw life support showed significantly better knowledge about BSD than did those who did not agree. Concerns about doctors' inclination to diagnose BSD to save resources and extract organs for transplantation were not observed to negatively affect subjects' decisions about life support withdrawal. CONCLUSION: The level of knowledge is an important factor affecting an individual's decision concerning withdrawal of life support therapy upon the diagnosis of BSD. Adequate explanation and counseling are important to facilitate family members in coping with this important end-of-life issue. PMID- 19545659 TI - Attitudes to brain death and organ procurement among university students and critical care physicians in poland. AB - PURPOSE: The practice of retrieving vital organs from brain-dead heart-beating donors is legally and medically accepted in Poland, but public beliefs and opinions regarding these matters have not been sufficiently explored. The purpose of this study was to evaluate the attitude of university students to the concepts of brain death and organ retrieval, compared with the attitude of critical care physicians. METHODS: The cohorts of 989 students and 139 physicians completed a questionnaire based on a survey instrument developed in an earlier reported study on Ohio residents. Participants assessed 3 scenarios: (1) brain death, (2) coma, and (3) vegetative state. More than 48% of students classified the patient from the brain death scenario as alive, and 51% of them were willing to donate organs of this patient. Ninety percent of students classified the patients in coma and in a vegetative state as alive, but still 34% of them would donate organs of those patients. The group of physicians properly determined the patients' diagnoses, but 10% of them accepted organ procurement from patients in coma and in a vegetative state. CONCLUSION: Our results supported the earlier observations of low public knowledge and inadequate understanding of brain death criteria and organ procurement processes. The majority of students were willing to accept organ procurement from severely ill but alive patients, in contrast with physicians. A considerable increase in public educational activity in this field is urgently recommended. PMID- 19545660 TI - Survey of medical students of Shahed University in Iran about attitude and willingness toward organ transplantation. AB - BACKGROUND: Transplantation is a successful procedure that prolongs the lives of people suffering from debilitating diseases. The purpose of this study was to assess medical students' attitude toward organ donation. METHODS: The study population consisted of 262 medical students with a mean age of 22.1 +/- 2.5 years who were surveyed using a reliable questionnaire that examined their attitudes and willingness. All data analyses were performed using Chi-square and analysis of variance tests with SPSS. RESULTS: The medical students had highly positive attitude toward organ donation (mean score 4.34 +/- 0.46) and a great willingness. Participants were more willing to donate their own organs than those of a deceased relative (85% vs 49.2%) to help others more than to develop science (91.2% vs 8.8%). The greatest willingness among students was kidney (84%) donation. There was no correlation between age, gender, education levels, and attitude toward organ donation. CONCLUSION: These findings necessitate an organized education program of medical students in all aspects of organ and tissue donation. PMID- 19545661 TI - Organ and tissue donation knowledge among intensive care unit nurses. AB - INTRODUCTION: Increased cadaveric donation may be achieved by improving medical staff education to enhance consent rates. This study was designed to examine the critical items that influence nurse knowledge regarding the concepts of brain death and the attitudes toward cadaveric donor renal transplantation. These findings may be useful to evaluate future progress in education. SUBJECTS AND METHODS: A 31-item questionnaire was distributed to 418 nurses in 24 intensive care units (ICUs) in our hospitals. In addition to initial data regarding age, sex, and marital status, knowledge about donation was evaluated by eight questions. The first four questions pertained to judicial and legal knowledge, the second two to economic knowledge, and the last two to concepts of brain death. RESULTS: Three hundred and two women and 116 men participated in this study. Their mean age was 31.98 years (range = 22-55 years). Two hundred fifty six (61.2%) of them were married. Three hundred sixteen (75.6%) expressed favorable attitudes regarding cadaveric donation. The major reason for their consent was humanity, and the major reason for their disagreement was "body respect." Of the 418 nurses, 105 (25.1%) stated positive attitudes toward living kidney donation. CONCLUSION: The legal definition of death and the medical definition of death should be uniform in the view of society. To make all medical staff favor cadaveric renal transplantation, they should agree on a definition of death. It was observed that the relatively high rate of misinterpretation of the brain-death concept among respondents negatively affected nurses attitudes toward cadaveric renal transplant. It was concluded that the "concept of brain death" and the "preferences of transplant over other treatment modalities" are the two items that need further attention in educational programs for nurses. PMID- 19545662 TI - Quality of life, coping, and mental health status after living kidney donation. AB - The main aim of the study was to explore well-being after donation. This retrospective, cross-sectional study of 161 living kidney donors (104 women; response rate 81.4%) who were aged between 32 and 80 years (x = 56.3; standard deviation = 10.9) included responses to standardized questionnaires concerning quality of life (QOL), coping, and mental health status. Most donors recovered fully from donation within 6 months (90.8%). Donor willingness to donate again (96.1%) was high. Their relationship to the recipient did not change (67.9%) or even improved (27.5%) in most cases. Donor QOL (as assessed by the Short-Form [SF]-36) did not differ from healthy norms. In 25% of donors, screening with the Hospital Anxiety and Depression Scale revealed anxious and/or depressive symptoms above the clinical cutoff score. Donor predominant coping style with their recipient's renal disease was "active problem-focused coping." The component scores of the SF-36 correlated positively with sociodemographic and self-reported medical parameters, coping, and mental health status. Although living kidney donation again proved to be a treatment without negative impact on donor QOL, the results underlined the importance of screening for donor mental health status and coping both in the evaluation process and after the procedure. PMID- 19545664 TI - Pretreatment with calcitonin gene-related peptide attenuates hepatic ischemia/reperfusion injury in rats. AB - OBJECTIVE: Oxygen free radicals and apoptosis play important roles in liver ischemia/reperfusion (I/R) injury. We sought to investigate the protective effect of calcitonin gene-related peptide (CGRP) to attenuate liver I/R injury due to oxygen free radicals and apoptosis. MATERIALS AND METHODS: Harvested rat livers were perfused via the portal vein with 60 mL of 4 degrees C histidine-tryptophan ketoglutarate (HTK) solution alone in the control group, or with the same solution containing CGRP (3 microg/10 g body weight) in the experimental group. After 24 hours of cold storage, hepatic enzyme leakage, portal venous pressure, oxygen consumption, total adenine nucleotides (TAN), bile production, lipoperoxide (LPO) release, apoptosis, and histochemical changes were evaluated upon 45 minutes of isolated reperfusion. RESULTS: Compared with control livers, CGRP-treated organs showed significantly decreased alanine aminotransferase (ALT) and glutamate-lactate dehydrogenase (GLDH) leakage and portal venous pressure (2.0 +/- 0.3 vs 4.0 +/- 0.4 mmHg; P < .01), with significantly increased bile production (8.56 +/- 0.76 vs 3.34 +/- 0.68 microL/g/45 min; P < .01), oxygen consumption (5.14 +/- 0.4 vs 2.57 +/- 0.2 microL/g/min; P < .01), and total adenine nucleotides (TAN) (11.1 +/- 0.71 vs 7.02 +/- 0.53 micromol/g; P < .01) upon reperfusion as signs of recovered viability. We observed infrequent positive terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling (TUNEL) staining, especially in sinusoidal lining cells (SLC). The percentage of TUNEL-positive cells in the CGRP group was significantly decreased compared with the control group: (4.1 +/- 0.67 vs 8.0 +/- 1.27; P < .05). Perfusate levels of low molecular weight (LMW) histone-associated DNA fragments (0.36 +/- 0.04 vs 0.53 +/- 0.06 AU; P < .05) were also decreased, coupled with strong 5' nucleotidase (5'-NT) and LDH activity staining concentrated on the endothelial cells. LPO release in the perfusate was largely decreased: (0.12 +/- 0.02 vs 0.36 +/- 0.04 nmoL/g, P < .01). CONCLUSION: CGRP ameliorated liver I/R injury due to reactive oxygen species and apoptosis. PMID- 19545663 TI - Addition of adenosine to University of Wisconsin solution: does it help? AB - Adenosine Triphosphate (ATP) precursors are sometimes added to preservation solutions in the belief that once the organ is reperfused, these precursors will build up ATP rapidly, returning it to its original metabolic state. This work studied ATP and metabolites during preservation of the rat liver using University of Wisconsin solution (UW), which contains adenosine, versus histidine tryptophan ketaglutarate solution a new phosphate-based preservation solution, or leeds solution (LS), which is under development at our institution (neither of the latter 2 contains adenosine). Tissue samples of perfused livers were analyzed for ATP and metabolites by high-performance liquid chromatography. UW did initially show the expected significant difference in overall adenosine levels, but the advantage had disappeared by 4 hours. At no time did UW show significantly higher levels of ATP; this was not seen following adenosine addition to LS. Only in living donor transplants where the cold ischemic time is short may there be some advantage to the addition of adenosine. PMID- 19545665 TI - Alleviating the ischemia-reperfusion injury of donor liver by transfection of exogenous hTERT genes. AB - BACKGROUND: Ischemia-reperfusion (I/R) injury had been linked to primary graft dysfunction in transplantation. To find effective methods to alleviate donor liver injury from I/R, we transferred exogenous human telomerase reverse transcriptase (hTERT) genes into donor rats before liver transplantation. METHODS: SD rats (age, 16 months) were divided into 3 groups: group A were donors pretreated with exogenous hTERT gene; group B were donors pretreated with adenovirus vector only; and group C were donors pretreated with physiologic saline. Alanine aminotransferase (ALT), apoptotic index, telomerase activity, and histological evaluation were calculated after liver transplantation. RESULTS: The levels of ALT and apoptotic index of group A were significantly lower than those of group B or group C (P < .05), at the same time, a mild histological injury and increased telomerase activity were also observed in group A. CONCLUSIONS: Exogenous hTERT gene provides protection against I/R injury, which depends on exogenous hTERT gene-mediated inhibition of apoptosis. PMID- 19545666 TI - Ex vivo transfer of nuclear factor-kappaB decoy ameliorates hepatic cold ischemia/reperfusion injury. AB - Cold ischemia/reperfusion injury of the hepatic graft has been attributed to the release of various inflammatory cytokines. Specific inhibition of these cytokines may improve viability of the hepatic graft upon reperfusion. Herein we have assessed the efficacy of cis element decoy against nuclear factor-kappaB binding site delivery to the hepatic tissue in a rodent liver transplantation model. At 8 hours after reperfusion of the liver, significant reduction was noted in the livers treated with decoy in the release of cytosolic enzymes from the hepatocytes and in serum tumor necrosis factor alpha (P < .05). The neutrophilic infiltration into the hepatic grafts was significantly suppressed in the livers treated with decoy oligodeoxynucleotides (ODNs). Decoy ODNs against nuclear factor-kappaB binding site delivery improved the viability of the hepatic graft against cold ischemia/reperfusion injury in the rodent liver transplantation model. PMID- 19545667 TI - Matrix metalloprotease expressions in both reperfusion lung injury and oleic acid lung injury models and the protective effects of ilomastat. AB - OBJECTIVE: Our aim was to study the expressions of matrix metalloprotease 9 (MMP9) and the effects of the MMP inhibitor Ilomastat in both ischemia/reperfusion (I/R)- and oleic acid (OA)-induced lung injury models. MATERIALS AND METHODS: Real-time polymerase chain reactions and Western blots were used to assess mRNA and protein expressions of MMP9 in lung tissues after I/R or OA lung injury. Ischemia was induced by clamping one branch of the pulmonary artery for 60 minutes and then reperfusing for 120 minutes. In the OA model, lung injury was induced by intravenous infusion of OA (0.1 mL/kg) for 20 minutes and then observation for 6 hours. Lavage leukocyte concentration and wet/dry lung weight ratio were used to assess lung inflammation and injury. Blood samples were collected for assays of hydroxyl radicals and nitric oxide. The MMP inhibitor Ilomastat (100 microg/kg) was administered before I/R and OA infusion. RESULTS: mRNA and protein expressions of MMP9 were significantly increased in both lung injury models. Ilomastat decreased MMP9 mRNA and protein expressions but did not reach statistical significance. Blood concentrations of hydroxyl radicals and nitric oxide, wet/dry lung weight ratios, and lavage leukocyte concentrations were significantly higher in both experimental groups compared with the sham group (P < .001). Ilomastat significantly attenuated the extent of lung inflammation and injury induced by both I/R and OA. CONCLUSION: MMP may play a critical role in the lung injury induced by I/R and OA infusion. PMID- 19545668 TI - Serine protease inhibitor aprotinin ameliorates renal injury in a rat model of ischemia-perfusion injury. AB - BACKGROUND: Renal ischemia-reperfusion (I/R) injury may occur after renal transplantation, thoracoabdominal aortic surgery, and renal artery interventions. OBJECTIVE: To investigate the therapeutic effects of aprotinin on tissue protection against I/R injury in a rat model. N-acetylcysteine (NAC), a potent antioxidant, was also tested to assess the experimental model. MATERIALS AND METHODS: Twenty-four rats were categorized into 3 groups of 8 rats each: those receiving isotonic sodium chloride solution (control group); NAC, 150 mg/kg; and aprotinin, 40,000 KIU/kg. The animals underwent unilateral nephrectomy after 60 minutes of warm ischemia and 60 minutes of reperfusion of the kidney. Malondialdehyde, a lipid peroxidation marker, and antioxidant glutathione levels were measured in the kidney parenchyma. Tissue samples were obtained for histologic analysis. RESULTS: Compared with the control group, the NAC group demonstrated significantly low levels of malondialdehyde (P = .04) and high levels of glutathione (P = .01). At histopathologic analysis, less acute tubular necrosis (ATN) and cellular swelling was noted in the NAC group (P = .002 and P = .005, respectively). In the aprotinin group, histopathologic analysis revealed less tissue damage in terms of ATN (P < .001, cellular swelling (P < .001), and vacuolysis (P = .002). Compared with the NAC group, ATN (P = .01), vacuolysis (P = .04), and congestion (P = .05) were significantly less in the aprotinin group. CONCLUSIONS: Our results suggest that administration of aprotinin attenuates renal I/R injury. This observation has potential application for kidney preservation for transplantation, for aortic surgery, and for renal artery interventions by protecting cells from free radical damage. PMID- 19545669 TI - Association between chronic hepatitis C infection and coronary flow reserve in dialysis patients with failed renal allografts. AB - BACKGROUND: Hepatitis C infection occurs frequently among patients with end-stage renal disease and increases the risk of atherosclerotic cardiovascular diseases. Endothelial dysfunction (ED) is an early event in the pathogenesis of atherosclerosis. It has been reported among patients treated with hemodialysis (HD), peritoneal dialysis (PD), or renal transplantation. The aim of the present study was to evaluate effects of chronic hepatitis C infection on ED in patients with failed renal transplants. METHODS: Twenty-six nondiabetic, anti-hepatitis C virus (HCV)-positive (15 females, mean age: 38 +/- 8 years) and 26 anti-HCV negative patients (15 females, mean age: 36 +/- 5 years), all of whom had returned to PD or HD after renal transplant failure were studied to assess coronary flow reserve (CFR) by transthoracic Doppler echocardiography. Serum high sensitivity C-reactive protein (hs-CRP) levels were measured as markers of chronic inflammation. CFR recordings and intima-media thickness measurements were performed using the Vivid 7 echocardiography device. RESULTS: Demographic and clinical characteristics of patients were similar between the two groups. Serum hs-CRP levels were significantly higher among HCV-positive patients versus HCV negative counterparts. HCV-positive patients showed lower CFR measurement than HCV-negative ones. Also, a negative correlation was observed between serum hs-CRP levels and CFR values. CONCLUSION: CFR values are worse among anti-HCV-positive patients with failed renal transplants compared with anti-HCV-negative subjects. Graft dysfunction per se may aggravate a proinflammatory states thereby inducing ED. Furthermore, the presence of HCV is a greater trigger of ED among patients with renal failed grafts. PMID- 19545670 TI - Informed consent in high-risk renal transplant recipients. AB - Before performing a clinical, diagnostic, and/or therapeutic action, the doctor is required to provide the patient with a bulk of information defined as informed consent. This expression was used for the first time in 1957 during a court case in California and the two words--informed and consent--are used together to underline the fact that the patient cannot give his or her true consent without first receiving correct information concerning the medical act in question. With regard to the medicolegal aspects governing organ transplants, despite the bulk of detailed work performed by health service workers involved in this surgical field with the aim of preparing adequate informed consent models, this has not yet been accompanied by the necessary legislative development. The informed consent model to be presented to the kidney transplant candidate should include a detailed description of the recipient's comorbidity and should aim at reducing the number of medicolegal actions, which have become more and more frequent in the last few years due to the ever increasing number of patients considered as suitable for transplantation. Informed consent, therefore, should not be a mere bureaucratic formality to be obtained casually, but should be carefully stipulated together with the patient by the transplant surgeon. It is, in fact, an indispensable condition for transforming a potentially illegal action, that is, the violation of an individual's psychophysical integrity, into a legal one. PMID- 19545671 TI - The role of Foxp3+ regulatory T cells in kidney transplantation. AB - Our project aimed to investigate the relation between the level of pretransplantation and posttransplantation peripheral CD4(+)CD25(+)Foxp3(+) T regulatory lymphocytes (Tregs) and the development of acute rejection (AR) episodes in 44 patients after kidney transplantation. During the 6-month period following transplantation, AR was diagnosed in 11 patients. Peripheral blood samples were collected 1 day before and 10 days after transplantation and tested for concentrations of CD4(+)CD25(+)Foxp3(+) cells by means of flow cytometry. The pretransplantation analysis showed significantly lower mean levels of peripheral Tregs in AR patients versus control group (P < .05). A lower level of Tregs was also observed in nonrejection (NONAR) patients versus control group (P < .05); however, it was still higher than in the AR group (P < .05). The 10-day posttransplantation analysis showed a similar pattern; however, a significant increase in the concentration of peripheral Tregs in NONAR patients was observed (P < .05), whereas no change was recorded in AR patients (P > .05). We found lower pretransplantation levels of peripheral Tregs in both AR and NONAR groups, versus control group. The deficiency of peripheral Tregs in patients with end stage renal failure might be due to the long-term inflammatory processes adversely affecting the peripheral regulatory mechanisms. However, significantly lower levels of Tregs observed in AR patients might also be related to genetic predispositions. Our observation suggests that the size and possibly the functionality of Tregs in the AR group was not sufficient to successfully control the immune response after kidney transplantation, leading to acute rejection episodes. PMID- 19545672 TI - Acute renal failure in the State of Qatar: presentation and outcome. AB - Despite major improvements in health care, acute renal failure is still one of the main prognostic factors in terms of patient mortality and long-term morbidity. This cohort prospective study to evaluate the patterns and outcomes of renal failure in Qatar was performed between January and June 2005. Of the 213 patients followed prospectively from referral to the end of their hospitalization, 66.7% were males and 33.3% females. Their overall mean age was 60 years; the majority were referred from critical care units. Comorbidity was present in 87% of all patients. Volume depletion, hypotension, and sepsis were the main predisposing factors for renal failure. Eighty three patients (39%) needed renal replacement treatment and 130 (61%) were treated conservatively. The majority of critical care patients needed dialysis. Overall mortality was 23.9%, 7% needed chronic dialysis, and 69.1% were discharged with normal or mild renal impairment. This study showed that acute renal failure was a major factor affecting patient mortality in Qatar. Early treatment of predisposing factors may improve overall patient outcomes. PMID- 19545673 TI - Kidney transplantation in children and adolescents: an analysis of United Network for Organ Sharing Database. AB - Specific pediatric allocation schemes can not only lead to minimization of waiting time, but also to better clinical outcomes for children with end-stage renal disease. The outcome of 4125 deceased donor kidney transplants (DDKT) aged 5-35 years were compared with those of 6456 living donor kidney transplants (LDKT) using univariate and multivariate Cox regression analyses. Unadjusted graft survival rates of DDKT were significantly lower than those of LDKT (hazards ratio [HR] = 1.53; P < .001). Chronic rejection was reported in 416 (10.1%) of 4125 in the DDKT group compared with 537 (8.3%) of 6456 in the LDKT group (P < .001). Among African American recipients, 67 (3.4%) grafts were lost due to noncompliance as a contributory cause of failure compared with 126 (1.5%) among other races (P < .001). A significantly lower incidence of noncompliance was observed in children (0.9%) compared with adolescents (2.2% in ages 10-14; P < .001) and high teens (2.0% in ages 15-20; P < .001). Multivariate analysis showed that adjusted graft survival rates of LDKT were superior to DDKT (HR = 1.22; P < .001) after adjusting for recipient race, recipient age, regraft status, and HLA mismatch. The differences of long-term graft survival rates between DDKT and LDKT have not been reduced (4% at 1 year, 10% at 3 years, and 12% at 5 years for unadjusted survival rates and 3% at 1 year, 6% at 3 years, and 9% at 5 years adjusted survival rates). In our analysis presented here the difference in graft survival between LDKT and DDKT has doubled compared with earlier analysis. Therefore, we recommend LDKT whenever possible as a first choice for pediatric transplant recipients. PMID- 19545675 TI - Outcomes of kidney grafts refused by one or more centers and subsequently transplanted at a single United Kingdom center. AB - INTRODUCTION: The rate-limiting factor in kidney transplantation is the shortage of donor organs with resulting steady increase in patients on the transplant waiting list. In our center we have seen an increase in the use of kidneys refused as unsuitable by one or more centers in the United Kingdom (UK). This study was performed to analyze the outcomes of transplantation from kidneys refused by one or more centers and subsequently transplanted by our institution. METHODS: We performed a retrospective analysis using the UK Transplant database of donor grafts refused by one or more centers and subsequently transplanted by us from January 2000 to December 2005. We documented the reason for refusal, donor and recipient factors, incidence of graft rejection, and primary and delayed graft function. Graft function and patient survival at 3 years were compared with standard donor grafts. RESULTS: From January 2000 to December 2005, we performed 623 renal transplantations, including 60 (9.6% from donors who were refused by one or more centers and 402 "standard" donor grafts. The main reasons for initial refusal included: elderly donor 25% (median age, 61 years), better HLA match required 33.3%, anatomical 5%, medical history of donor 6.6%, virology 4.8%, prolonged cold ischemia time 3.3% (median, 33.5 hours), and organ damage 1.6%. The 3-year median creatinine levels of donor grafts refused by multiple centers was 126 mumol/L compared with 135 mumol/L for standard grafts (P = .97). Three-year graft and patient survival rates were 86.6% and 96%, for grafts refused by multiple centers and 87% and 95%, for standard grafts, respectively. Upon multivariate analysis none of the above variables were significant predictors of 3-year failure of grafts refused by multiple centers. CONCLUSIONS: Nearly 10% of kidney transplants in our center were performed with grafts refused by one or more centers as "unsuitable." The graft and patient survivals were similar to those of standard grafts. None of the factors for refusal of kidneys by other centers predicted graft failure at 3 years. There may be an element of subjective assessment and subsequently a "cascade effect" involved in refusal of some of these kidneys. PMID- 19545674 TI - Expression of carcinoembryonic antigen-related cell adhesion molecule 1 in acute rejection of human renal allografts. AB - BACKGROUND: Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is expressed on various cell types and mediates homophilic cell adhesion. CEACAM1 plays an important role in cell morphogenesis and angiogenesis. Furthermore, CEACAM1 regulates adhesive activity of immune-competent cells, suggesting an additional role in inflammatory processes. METHODS: Therefore, in the present study the expression of CEACAM1 was analysed retrospectively in renal biopsies from kidney transplant recipients (stable graft [Ctr; n = 18], acute vascular rejection [AVR; n = 14], acute tubulointerstitial rejection [AIR; n = 9], and combined vascular and interstitial rejection [AVIR; n = 7]). Expression patterns of CEACAM1 were determined using immunohistochemistry and quantitative morphometry. RESULTS: All biopsy specimens from patients with stable grafts showed low CEACAM1 levels, suggesting a constitutive expression in renal transplants. In patients with acute rejection, CEACAM1 was markedly up-regulated. AVR revealed the highest tubular CEACAM1 levels (4.9 +/- 0.5% [AVR] vs 2.2 +/- 0.3% [Ctr] of tubular area; P < .05), whereas interstitial rejections showed the highest glomerular expressions (4.5 +/- 0.5% [AIR] vs 0.9 +/- 0.1% [Ctr] of glomerular area; P < .05). CONCLUSIONS: An up-regulated expression of CEACAM1 in tubular and/or glomerular cells is an indicator of acute inflammatory processes in biopsy specimens from patients with acute renal allograft rejections and, therefore, might be used as a new clinical marker. PMID- 19545676 TI - Relationship between platelet activation and acute rejection after renal transplantation. AB - OBJECTIVE: To provide an earlier diagnosis and efficiently treat acute rejection episodes (ARE) after renal transplantation, we studied its relationship to platelet activation. PATIENTS AND METHODS: The peripheral blood levels of platelet surface glycoprotein (CD61), platelet activation-dependent granule membrane protein (CD62p), lysosomal enzyme glycoprotein (CD63), macula densa granule membrane glycoprotein (CD42a), and fibrinogen receptor monoclonal antibody (PAC-1) among 203 patients with uremia in various stages before and after transplantation were assayed by flow cytometry. The patients with ARE were prospectively and randomly assigned to either a treatment group with an antiplatelet activation agent or a control group. RESULTS: The incidence of ARE was remarkably increased among patients with greater expression levels of platelet activation markers in peripheral blood preoperatively. The values of platelet activation markers were significantly higher among patients with ARE compared with those showing either normal graft function or acute tubular necrosis. The greater the increase in CD63, the worse the ARE. The expression levels of platelet activation markers decreased remarkably among the group treated with a platelet activation inhibitor in addition to antirejection therapy: the rejection reversal time shortened and the dose of antihuman thymocyte globulin (ATG) was lower. The sensitivity of platelet activation markers was better than its specificity. CONCLUSIONS: Our studies demonstrated an association between platelet activation and ARE after renal transplantation. Platelet activation before transplantation can predict the occurrence of ARE. Platelet activation inhibitor therapy after transplantation improved ARE reversal. PMID- 19545677 TI - Abnormal high expression of B-cell activating factor belonging to the TNF superfamily (BAFF) associated with long-term outcome in kidney transplant recipients. AB - B-cell activating factor belonging to the tumor necrosis factor superfamily (BAFF) is a critical regulator of B-cell maturation and survival. We investigated the expression of BAFF in peripheral blood sample from kidney transplant recipients. Results of flow cytometry showed that at 5 years or more posttransplantation, cell-surface BAFF was significantly expressed on peripheral CD3+ T lymphocytes kidney transplant recipients in compared with other groups (P < .05). BAFF expression was noted on CD4+ and CD8+ T cells. The BAFF messenger RNA level in peripheral mononuclear cells was consistent with the protein level. However, the serum soluble BAFF level varied among individuals in each group. Stratified by renal function, the cell-surface BAFF level was significantly higher in recipients with abnormal renal function compared with those with normal renal function (P < .05). Enzyme-linked immunosorbent assay results showed that expression level of cell-surface BAFF significantly correlated with panel reactive antibody. These results indicate that BAFF may be involved in the development of graft loss. PMID- 19545678 TI - Effect of CYP3A5 genotype on renal allograft recipients treated with tacrolimus. AB - OBJECTIVE: Tacrolimus concentrations are associated with CYP3A5 genotype. The purpose of this study was to evaluate the outcomes and drug concentrations/doses among a posttransplant population with various CYP3A5 genotypes within 12 months. METHODS: Sixty seven kidney recipients receiving immunosuppression with tacrolimus + mycophenolate mofetil + prednisolone were grouped according to their CYP3A5 genotypes (*1/*1; *1/*3; *3/*3). The initial dose of tacrolimus (0.15 mg/kg/d) was adjusted according to achieve a target therapeutic window. All patients underwent a protocol biopsy at 1 month posttransplantation. We assayed serum creatinine and tacrolimus blood trough concentrations to calculate the concentration per dosage during follow-up. We also investigated the incidence of acute rejection episodes and the nephrotoxicity of tacrolimus according to the renal biopsy. RESULTS: There was no significant difference among serum creatinine concentrations. Tracrolimus blood concentrations showed a significant difference at day 7 and 1 month with no significant difference at 3, 6, or 12 months among the three groups. The CYP3A5*3/*3 group showed the largest concentration per dosage (C/D) and CYP3A5*1/*1, the smallest C/D. There was a significant difference among the three groups. The occurrence of an acute rejection episode within 3 months showed a significant difference among the three groups but not from 3 to 12 months after transplantation. Nephrotoxicity was greatest among the CYP3A5*3/*3 group. CONCLUSION: CYP3A5 influenced the blood concentrations of tacrolimus. Our study suggested to choose the initial dosage according to the CYP3A5 genotype to obtain a better outcome and reduce the incidences of acute rejection episodes and nephrotoxicity. PMID- 19545679 TI - Interleukin-17 and kidney allograft outcome. AB - Acute rejection episodes (ARE) are important complications that involve the interplay between mechanisms that maintain graft tolerance and promote rejection. The proinflammatory cytokine interleukin-17 (IL-17) has been implicated in many conditions in humans and mice. In kidney transplant patients, the evaluation IL 17 levels has been performed in only a few patients. We performed a cross sectional study correlating quantitative IL-17 levels and clinical outcomes. PATIENTS AND METHODS: We studied 19 specimens from biopsies performed in patients (n = 19) who received isolated kidney grafts. ARE signs were present in 9 (47%) patients who provide specimens; whereas, 10 (53%) others showed no signs of rejection. Eighteen healthy control sample IL-17 underwent measurement, all of which were performed by an enzyme-linked immunosorbent assay method. We assessed other factors, such as the recipients demographic data, cold ischemia time, HLA mismatches, time elapsed from transplantation to the biopsy, posttransplantation status, antibody panel, donor type, and immunosuppressive treatment. RESULTS: IL 17 levels were clearly increased among samples derived from patients with ongoing rejection (125.7 +/- 27.06 pg/mL) in contrast, to the nonrejection group, (30 +/- 13.32 pg/mL) (P < .05). Healthy controls showed no detectable IL-17 levels. CONCLUSIONS: These findings suggested that IL-17 was important in the pathophysiology of acute kidney rejection. PMID- 19545680 TI - Clinical application of Cordyceps sinensis on immunosuppressive therapy in renal transplantation. AB - OBJECTIVE: We sought to explore the adjunctive effects of Cordyceps sinensis (CS) in clinical renal transplantation. MATERIALS AND METHODS: Patients (n = 202) were divided randomly by lottery into a treatment (n = 93) and a control group (n = 109). Patients in the treatment group were treated with CS 1.0 g 3 times a day in addition to the immunosuppressive regimen given to the control group. We compared patient and graft survivals, incidence, time and severity of acute rejection episodes, chronic allograft nephropathy (CAN), hepatotoxicity and nephrotoxicity, biochemistry parameters including indicators of liver and kidney functions, fats, proteinuria, dosages, and whole blood concentrations of cyclosporine (CsA). RESULTS: Patient and graft survival rates, serum creatinine (SCr), and blood urea nitrogen (BUN) were not significantly different between the 2 groups (P > .05). Serum uric acid (UA) and 24-hour urinary total protein (24-hour UTP) were significantly lower in the treatment group than in the control group (P < .05). The incidences (11.83% vs 15.60%) and times to acute renal allograft rejection (23.48 +/- 7.22 vs 22.27 +/- 8.03 days posttransplantation) were not significantly different between the treated and control groups (P > .05). Patients receiving thymoglobulin antirejection therapy (3 cases) were fewer in the heated versus control group (13 cases; P = .014). The incidences of hepatotoxicity and nephrotoxicity in the treated group were 12.90% and 19.35%, significantly lower than 24.77% and 33.94% in the control group, respectively (P < .05). At 2 to 6 months posttransplantation, the CsA dosages in the treated group were significantly lower than those in the control group (P < .05). The whole blood trough CsA concentrations in the treated group were significantly lower than those in the control group at 3 to 6 months posttransplantation (P < .05). The decreasing trends of the 2 aforementioned parameters in the treatment group were approximately linear among treated subjects compared with approximately quadratic in the control group (P < .05). The incidence of CAN in the treated group was 7.53%, which was significantly lower than 18.35% in the control group (P = .024). The 24-hour UTP level in CAN patients within the treated group was significantly lower than the control group after transplantation (P = .045). The differences in total bilirubin, SCr, serum UA, and total cholesterol levels among otherwise normal patients in the treated group were significantly lower than those among the control group (P < .05). CONCLUSIONS: The use of CS may allow decreased dosages and concentrations of CsA causing fewer side effects without an increased risk of acute rejection. In addition, CS with reduced dose CsA may decrease proteinuria and retard CAN progression. PMID- 19545681 TI - Increased asymmetric dimethylarginine serum levels are associated with acute rejection in kidney transplant recipients. AB - Asymmetric dimethylarginine (ADMA) has been identified as a marker of endothelial dysfunction and an independent risk factor for cardiovascular events in uremic subjects. This study evaluated ADMA plasma levels in kidney transplant recipients. ADMA levels were serially measured during the first year posttransplantation in 41 recipients treated with cyclosporine regimen (CY), sirolimus (SIR), or low-dose cyclosporine plus everolimus (E). Homocysteine, C reactive protein (CRP), nitric oxide (NO), and standard routine laboratory analyses were determined serially. ADMA significantly increased at 6 months posttransplantation, but was significantly lower among patients on SIR or E. NO was only slightly reduced in patients with increased ADMA levels. Interestingly, ADMA was significantly increased during the first 4 days posttransplantation in patients who experienced acute rejection during the first 6 months after transplantation. The same group of patients demonstrated higher levels of CRP and systolic blood pressure before transplantation. Our results demonstrated that ADMA was increased in patients on CY at 6 months. When increased soon after transplantation ADMA may be associated with episodes of acute rejection in kidney transplant recipients. The presence of elevated systolic blood pressure, as well as CRP and ADMA levels, suggested a role for endothelial dysfunction in the development of acute rejection episodes among deceased donor kidney transplant recipients. PMID- 19545682 TI - Antigen-specific suppression by induced CD4+CD25high regulatory T cells in kidney recipients. AB - The biology and function of induced CD4(+)CD25(high) regulatory T (Treg) cells have not been clarified for their specificity to a foreign antigen. To test whether the regulatory functions of the induced CD4(+)CD25(high) Treg cells after transplantation require antigen-specific triggering, we analyzed the capacity of induced CD4(+)CD25(high) Treg cells to inhibit the proliferation of conventional CD4(+)CD25(-) T cells in response to T-cell receptor stimulation using donor cells or HLA-mismatched third-party cells in vitro. CD4(+)CD25(high) Treg cells did not proliferate in response to allogeneic stimulation and suppressed proliferation of the co-cultured autologous CD4(+)CD25(-) populations in a dose dependent manner. The proliferation of CD4(+)CD25(-)T cells from the same donor in mixed lymphocyte reactions was significantly inhibited at a 1:8 ratio of conventional T cells:Treg cells: 14,404 +/- 673 cpm without CD4(+)CD25(high) Treg cells versus 10,781 +/- 539 cpm with CD4(+)CD25(high) Treg cells P = .01). At the same 1:8 ratio, the proliferation of CD4(+)CD25(-) cells derived from major histocompatibility complex-mismatched patients was not significantly inhibited: 14,404 +/- 673 cpm without CD4(+)CD25(high) Treg cells versus 12,471 +/- 709 cpm with CD4(+)CD25(high) Treg cells (P = .06). Antigen specificity of the induced CD4(+)CD25(high) Treg cells was demonstrated, after transplantation, supporting the use of antigen-specific Treg cells as a therapeutic strategy. PMID- 19545683 TI - A multiplex bead array analysis to monitor donor-specific cytokine responses after withdrawal of immunosuppression in HLA-identical living related kidney transplant patients. AB - Immune reactivity after HLA-identical living related (LR) kidney transplantation can be caused by minor histocompatibility antigen and non-HLA antigen mismatches between donor and recipient. In our center, HLA-identical LR kidney transplant recipients receive azathioprine (AZA) or mycophenolate mofetil (MMF) in combination with corticosteroids for 1 year after transplantation. Thereafter, AZA or MMF was withdrawn, and the patients were treated with steroid monotherapy as maintenance therapy. We questioned whether withdrawal of AZA or MMF affected the donor-specific lymphocyte proliferation and cytokine production. Donor and third-party T-cell reactivities were determined by mixed lymphocyte reactions and by cytokine production using multiplex bead array technique. The donor and third party proliferative capacities were not affected after withdrawal of AZA or MMF. Thirteen of 17 cytokines were detected by the multiplex bead array technique. No differences were observed after third-party induced cytokine production after withdrawal of AZA or MMF. However, production of donor-specific interferon-gamma and macrophage inflammatory protein-1beta increased after discontinuation of AZA or MMF, but no clinically relevant acute rejection was observed. In conclusion, after HLA-identical LR kidney transplantation, donor-specific cytokine responses can be found when AZA or MMF therapy is discontinued. The clinical relevance of this phenomenon is still not evident. PMID- 19545684 TI - Trends in kidney transplantation outcome: the Andalusian Kidney Transplant Registry, 1984-2007. AB - Herein we have presented the first report from the Andalusian Kidney Transplant Registry, a Public Health Service Regional Registry in Andalusia, Spain (general population, 8 million). The current analysis was limited to 5599 kidney-alone transplants from deceased donors, grouped into 4 time periods: 1984-1989 (n = 846); 1990-1995 (n = 1172); 1996-2001 (n = 1801); and 2002-2007 (n = 2060). The age of the transplant patients rose over time to 21.7% of recipients of ages >or=60 years in 2002-2007. In the later years we observed an increased incidence of vascular and diabetic causes of end-stage renal disease (ESRD). Patients who underwent retransplantation increased from 2.7% in 1984-1989 to 8.1% in 2002 2007. Time on previous renal replacement therapy (RRT) increased from 33.1 +/- 29 to 48 +/- 53 months. Patient survivals at 1, 5, 10, and 20 years were 96%, 91%, 83%, and 63%, respectively. Censoring for death, graft survivals were 90%, 80%, 67%, and 45%, respectively. Compared with the 1984-1989 period, patient survival improved by about 10% (P < .001) since 1990, remaining stable to 2007. Censored 5 year graft survivals progressively improved from 72% to 77%, 82%, and 85% (P < .001). Upon multivariate analysis, gender, age >39 years, diabetes, and RRT duration were independent predictors of patient survival. Age <18 years, retransplantation, and positive hepatitis C virus serology were independent predictors of lower graft survival. Considering 1984-1989 as the reference time period, both patient and graft mortality risks continuously decreased over the following 3 periods (relative risk [RR] = 0.5-0.4-0.3 for patient mortality; RR = 0.8-0.6-0.5 for graft mortality). In summary, despite an increased number of adverse risk factors, both patient and graft survivals have improved from 1984 to date. PMID- 19545685 TI - Pregnancy in renal transplant recipients. AB - OBJECTIVE: Renal transplantation with a well-functioning graft leads to a rapid restoration of endocrine and sexual functions. The aim of this study was to examine our experience with pregnancies among renal transplant patients, particularly with regard to their impact on graft function. PATIENTS AND METHODS: We analyzed 10 pregnancies in 7 renal transplant recipients for long-term graft outcomes in terms of clinical and biological data. RESULTS: The mean patient age was 28.5 +/- 4 years. They all received a living donor kidney. The time between transplantation and the onset of pregnancy was 33.4 +/- 23.2 months. Regarding the immunosuppressive therapy, all patients received steroids and cyclosporine; 4 patients received in addition azathioprine and 2 received mycophenolate mofetil that was changed at 1 month before conception to azathioprine. There was no significant difference between the serum creatinine before and during pregnancy. We did not observe any acute rejection episode. Pregnancy complications were preclampsia in 1 case, hypertension in 1 case, urinary tract infection in 2 cases, and anemia in 80% of patients during the third trimester. Premature rupture of membranes occurred in 1 case and preterm delivery in 2 cases. Two cases of neonatal death were registered. Cesarean section was performed in 50% of cases. The follow-up revealed 2 cases of chronic rejection. CONCLUSION: A multidisciplinary approach is necessary for pregnancy which generally occurs at 2 years after kidney transplantation. PMID- 19545686 TI - Association between Toll-like receptors 4 and 2 gene polymorphisms with chronic allograft nephropathy in Turkish children. AB - Toll-like receptor (TLR) gene polymorphism is known to impair intracellular signaling pathways following adaptive immune responses. Our aim was to investigate the distribution of TLR4 and TLR2 gene polymorphisms among pediatric renal transplantation patients in relation to chronic allograft nephropathy (CAN). In addition to 115 healthy controls, we included 69 renal recipients, 19 of whom were identified as CAN by biopsy scored according to the Banff criteria. Polymorphisms at TLR4 Asp299Gly and/or Thr399Ile were present in 11.6% of renal transplant recipients. None of these subjects was identified in cosegregation with the Thr399Ile allele, whereas three had an isolated Asp299Gly and five had an isolated Thr399Ile. Neither renal recipients nor healthy controls were homozygous for both Asp299Gly and Thr399Ile polymorphisms. However, TLR4 Thr399Ile polymorphism and Ile allele was greater among CAN (-) versus CAN (+) recipients (P > .05). The frequency of TLR2 mutant type Gln allele was significantly higher in recipients than among healthy controls (P < .0001). However, the Gln allele frequency was similar between CAN (+) and CAN (-) patients. The results of present study may be speculated to show TLR4 and TLR2 gene polymorphisms as protective factors from CAN development due to impaired immune responses. PMID- 19545687 TI - Risk factors for graft dysfunction in the late period of renal transplantation. AB - INTRODUCTION: Risk factors traditionally associated with kidney graft dysfunction after transplantation are delayed graft function, acute cellular rejection episodes, deceased donor organ source (particularly more than 50 years old), and HLA mismatch. Socioeconomic factors, such as income, education, and type of health insurance, have been reported in some studies. OBJECTIVE: To evaluate the risk factors traditionally associated with worse function of the transplanted kidney and the role of socioeconomic variables among our population. DESIGN OF STUDY: A cohort with 69 patients transplanted in the period 2003 to 2006 was assessed for predictors for a creatinine clearance estimated to be less than 60 mL/min or stage 3 or greater of renal dysfunction at 1 year of follow-up. CONCLUSION: Upon bivariate regression analysis, acute tubular necrosis [odds ratio (OR) 6.93 (1.9-24), P = .03], and bacterial infection [OR 4.13 (1.4-12), P < .01] were predictors of risk, which was also observed among transplants of kidneys from deceased donors [OR 2.67 (0.9-7.6), P = .07] or donors aged more than 49 years [OR 4.22 (0.9-18.1), P = .06]. Better education was a significant protective factor [OR 0.3 (0.1-0.9), P = .02]. Upon multivariate logistic regression analysis delayed graft function [OR 5.1 (1.3-20.5), P = .02] and severe bacterial infection [OR 3.9 (1.23-12.9), P = .02] were predictors; there was no association with socioeconomic factor. PMID- 19545688 TI - Significance of HLA nondependent risk factors of chronic transplant nephropathy for the development of endothelial dysfunction after kidney transplantation. AB - More than 40% of renal allografts show chronic transplant nephropathy (CTN) early after renal transplantation. Cardiovascular disease is the leading cause of death in this population. Thus endothelial dysfunction represents an early angiopathy causing CTN and atherosclerosis. We sought to evaluate changes in endothelial dysfunction and vascular wall thickness during the first year posttransplantation as well as their association with HLA nondependent risk factors for CTN. At 3 and 52 weeks after grafting, we studied 25 patients without overt atherosclerotic disease and acute posttransplant complications for von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator (tPA), big endothelin-1 (ET-1), flow-mediated dilatation (FMD), intimal media thickness (IMT), serum total cholesterol (TC), and triglycerides (TAG). FMD and IMT at 52 weeks showed significant correlations (P < .05) with recipient age, and the FMD ratios at 3 and 52 weeks correlated with the time on hemodialysis. Recipient age was significantly correlated with TC and PAI-1 with TAG. vWF was the only parameter that significantly correlated with donor age. There were no significant correlations with creatinine clearance. Decreased TAG approached statistical significance (P = .07) and TC decreased nonsignificantly. The worsening of FMD and ET-1 was not significant. A nonsignificant improvement in IMT was not associated with any analyzed parameters. Our results implied that the time on hemodialysis, the presence of hyperlipoproteinemia, and the recipient age significantly contributed to endothelial dysfunction during the first year after transplantation. PMID- 19545689 TI - Posttransplant proteinuria is associated with higher risk of cardiovascular disease and graft failure in renal transplant patients. AB - In this study, we sought to determine whether proteinuria after renal transplantation was associated with cardiovascular disease (CVD), patient survival, and long-term allograft survival. One hundred twenty-six patients included 102 males and 24 females of mean age 30.7 +/- 8.9 years. Their mean follow-up was 63.21 +/- 19.9 months. All patients were evaluated for CVD, namely, ischemic heart disease, cerebrovascular disease, and peripheral vascular disease. Proteinuria was defined as urinary protein >or=500 mg/d which persisted for >6 months after transplantation. We retrospectively examined pre- and posttransplant data, including sex, age at transplantation, smoking, pretransplant dialysis duration, donor status, number of acute rejection episodes, body mass index, systolic and diastolic blood pressure levels, lipid profile and other biochemical parameters, immunosuppressive regimens, as well as pulse steroid dose. Proteinuria was significantly associated with CVD (P = .001; RR = 6.43; confidence interval [CI] 2.15-19.22). Patients with proteinuria showed significantly lower graft survival rates than those without proteinuria (58.62% vs 80.41%; P = .02). The mean time to appearance of proteinuria was 14.1 +/- 11.4 months (range, 1-36 months). There was no significant association between proteinuria and patient survival. Patients with persistent proteinuria displayed a greater number of acute rejection episodes (1.20 +/- 1.17 vs 0.62 +/- 0.85; P = .004) and higher pulse steroid dosages (4380.0 +/- 3123.4 vs 2800.0 +/- 2766.7; P = .022). In conclusion, persistent proteinuria is a strong risk factor for CVD among renal transplant patients. Therefore, an etiologic search and antiproteinuric strategy should be considered routinely to improve patient and graft outcomes. PMID- 19545690 TI - Vascular complications in renal transplantation: a single-center experience in 1367 renal transplantations and review of the literature. AB - Renal transplantation is the treatment of choice for end-stage renal disease. Vascular complications in renal transplantation are not uncommon and may often lead to allograft loss. The most common vascular complications are transplant renal artery stenosis, transplant renal artery thrombosis, transplant renal vein thrombosis, biopsy-induced vascular injuries, pseudoaneurysm formation, and hematomas. Transplant renal artery and vein thrombosis have an early onset and a dramatic clinical manifestation and usually lead to allograft loss. In contrast, transplant renal artery stenosis has better treatment possibilities, whereas the rest do not occur so often. In our institution, 1367 renal transplantations were performed from September 1980 to April 2005. During this period, we encountered 38 major vascular complications leading to graft loss and 19 transplant renal artery stenoses with successful treatment in the majority of cases. According to these data, we can conclude that renal transplantation is a safe therapeutic procedure for renal failure. PMID- 19545691 TI - Postoperative voiding dysfunction in older male renal transplant recipients. AB - OBJECTIVE: To evaluate the incidence of voiding dysfunction in older male renal transplant recipients. PATIENTS AND METHODS: Data for 103 patients aged 60 years or older (mean age, 65.7 years; group 1) who underwent transplantation at our center between January 1999 and August 2007 were compared with data for a group of 139 younger patients (mean age, 50.1 years; group 2) treated within the same time frame. RESULTS: Postoperatively, 28 group 1 recipients (27%) and 26 group 2 recipients (19%) experienced voiding dysfunction after removal of the transurethral catheter (P = .12). The most common cause was bladder outlet obstruction due to benign prostatic hyperplasia in 26 patients in group 1 (25%) and 17 patients in group 2 (12%) (P = .009). Bladder neck contracture, urethral stricture, and detrusor underactivity were diagnosed in the other patients. Transurethral resection of the prostate gland was performed in 21 group 1 patients (20%) and 14 group 2 patients (10%) (P = .02) at a mean of 31.1 and 29.5 days, respectively (P = .23) after transplantation. Surgical procedures were performed without complication, and symptoms did not recur postoperatively. CONCLUSIONS: Our data reveal a high incidence of voiding dysfunction in older male renal transplant recipients. High residual urine and urinary retention after renal transplantation may induce recurrent urinary tract infections, cause relevant complications, and seriously affect graft function. Recognizing the substantial effects of postoperative voiding dysfunction will enable optimum management of older kidney transplant recipients. PMID- 19545692 TI - De novo urothelial carcinoma in kidney transplant patients with end-stage aristolochic acid nephropathy in China. AB - OBJECTIVE: Aristolochic acid nephropathy (AAN) is a progressive renal interstitial fibrosis disease that was initially reported among a Belgian cohort of about 50 patients after the intake of diet pills containing the Chinese herb Aristolochia fangchi. In addition to renal disease, foci of AAN show increased incidences of urothelial carcinomas (UC). Immunosuppression is associated with an increased risk for the development of different malignancies. Our aim was to examine the outcomes of UC among patients with AAN after transplantation in China, the cradle of this traditional medicine. PATIENTS AND METHODS: We performed a retrospective evaluation of the charts and pathology reports of 1612 renal transplant recipients treated at our 2 institutions. RESULTS: From January 1998 to December 2006, we performed cadaveric kidney transplantations in 17 patients with AAN, all of whom were treated with cyclosporine plus azathioprine or mycophenolate mofetil plus prednisone. One-year graft survival was 100%. During the mean follow-up of 57 months (range, 21-108 months), 9 recipients (52.9%) developed UC, compared with a 0.46% prevalence of urinary tract tumors among other Chinese kidney transplant recipients. The age at which the diagnosis was made ranged from 39 to 66 years (mean, 53.6 +/- 6.8 years). Among the 9 patients with UC, 8 cases (88.9%) involved the upper urinary tract: bilateral, 3 cases, 37.5%; unilateral, 5 cases, 62.5%. In 1 patient only a bladder tumor was detected. Two patients showed the bladder, synchronous bilateral ureter, and pelvis to be involved. All patients with UC underwent surgical treatment, recovering uneventfully with functioning grafts after tumor excision, except 1 patient who underwent nephrectomy of the transplanted kidney. Six patients (75%) experienced recurrences during the follow-up period. Three patients died within a mean of 20 months (range, 1-42 months) after tumor excision. CONCLUSIONS: The risk for UC is increased among patients with AAN after transplantation. Regular screening for early detection of malignancy is mandatory. Longer follow-up and results from other transplant centers are needed to further investigate the relationship between AAN and UC among renal transplant patients. PMID- 19545693 TI - Treatment of urinary fistula after kidney transplantation. AB - Urinary fistula is a common complication after kidney transplantation and may lead to graft loss and patient death. Its current incidence ranges from 1.2% to 8.9%. From December 1993 to April 2007, 1223 kidney transplant procedures were performed by our kidney transplantation team. In 948 recipients (group 1), we performed an extravesical ureteroneocystostomy, and in 275 recipients (group 2), a terminoterminal ureteroureterostomy (UU). We observed urinary fistulas in 43 patients (3.5%), with mean onset at 6 days (range, 3-20 days) posttransplantation. Urinary fistula was significantly more common in group 1 compared with group 2 (4.1% and 1.5%, respectively; P < .05). The distal ureteral necrosis was the major frequent cause of urinary fistula (n = 34; 76.7%), which required either a second ureteroneocystostomy or UU using the native ureter. Of these 21 fistulas, including 10 recurrent fistulaes, were successfully treated with pedicled omentum covering the anastomotic stoma. Conservative treatment with a stent and Foley catheter drainage for 1 to 2 weeks was successful in 8 patients. All patients with a urinary fistula regained normal graft function except 1 in whom transplant nephrectomy was necessary because of pelvic and ureteral necrosis. There was no recipient loss secondary to urinary fistula. In conclusion, UU can decrease the incidence of urinary fistula after kidney transplantation. Most urinary fistulas require surgical management; and pedicled omentum is useful to repair the fistula. PMID- 19545694 TI - Major depressive disorder in children and adolescents after renal transplantation. AB - This cross-sectional study evaluated the prevalence of major depressive disorder and depressive symptoms in children and adolescents after renal transplantation. A total of 71 patients who had undergone renal transplantation were interviewed in person using the Farsi (Persian) version of the Kiddie Schedule for Affective Disorders and Schizophrenia and Diagnostic and Statistical Manual of Mental Disorders diagnostic criteria. Major depressive disorder, depressive symptoms, and suicidal behaviors were assessed. The rate of major depressive disorder was 2.8%; two-thirds of the patients had irritability; and approximately 40% had recurrent thoughts of death and suicidal ideation. The rate of major depressive disorder was lower than in other chronic diseases such as thallasemia or hemophilia; however, the rate of suicidal behaviors was high. PMID- 19545695 TI - Low-density lipoprotein oxidizability and the alteration of its fatty acid content in renal transplant recipients treated with cyclosporine/tacrolimus. AB - Renal transplantation is widely used to treat patients with end-stage renal disease. Atherosclerosis is an important posttransplantation risk factor for renal transplant recipients. Subsequent to transplantation low-density lipoprotein (LDL) particles become susceptible to oxidative modification, which results in atherosclerosis. Therefore, the aim of our study was to investigate differences in the susceptibility of LDL particles to oxidation by analyzing LDL fatty acid levels among renal transplant recipients. The changes in lag phases and fatty acid levels of LDL were observed over 4 months among renal transplant recipients treated with Cyclosporine (CsA; n = 7) or Tacrolimus (FK-506; n = 9). We also analyzed cholesterol and triglyceride levels of patients and healthy controls. The lag phase at the 60th day after transplantation was significantly shorter than the results either before transplantation or among control subjects. In conclusion, a similar decrease in lag phase was observed in both above groups, but the FK-506-treated group showed a better lipid profile than the CsA-treated group. PMID- 19545696 TI - Role of uric acid in post-renal transplantation hypertension. AB - Uric acid (UA) is an emerging cardiovascular (CV) risk factor that is associated with hypertension and CV disease (CVD) in the general population, but whose role in renal transplant recipients (RTR) has not been defined. We performed a retrospective chart review of 589 stable RTR receiving ongoing posttransplant care at our hospital, identifying those with a minimum of 3 serum UA measurements obtained at least 2 months posttransplantation, 6 months graft survival, stable renal function, and no change in antihypertensive or immunosuppressive drugs over this time. Data were collected for the period November 2005 to July 2007. Relationships were assessed by Pearson's correlation coefficient, and correlates of UA including blood pressure (BP) were determined using multiple linear regression analysis. There were 464 RTR who met eligibility criteria for the study. Hyperuricemia was present in 196 patients (42%). By Pearson's correlation coefficient, UA was inversely correlated with estimated glomerular filtration rate (eGFR; R = -.39; P < .0001) and directly correlated with C-reactive protein (CRP; R = .10; P = .02). However, UA did not correlate with either age (R = .07; P = .08) or systolic BP (R = .05; P = .76). Upon multivariate linear regression, UA was inversely associated with eGFR (P < .0001) and directly associated with male gender (P < .0001), use of cyclosporine (CsA; P = .0002), increasing time posttransplantation (P = .007), and CRP (P = .01). In summary, hyperuricemia is common in RTR, but was not related to BP. Further studies are required to establish whether UA predicts CV risk in this population. PMID- 19545697 TI - Chronic renal disease in renal transplant patients: management of cardiovascular risk factors. AB - Kidney transplantation is the treatment of choice for patients with end-stage renal disease. Despite improvements in short-term patient and graft outcomes, there has been no major improvement in long-term outcomes. The aim of this study was to determine the prevalence of cardiovascular risk factors, such as hypertension, dyslipidemia, diabetes, chronic kidney disease, and obesity, and the impact of their control among 526 stable renal transplant recipients according to the guidelines in the general population. Mean blood pressure was 133 +/- 16/81 +/- 9 mm Hg. The proportion of patients on antihypertensive therapy was 75%, and on ACE inhibitors or angiotensin II receptor blockers, 26%. The mean cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and triglycerides were 195 +/- 41, 115 +/- 32, 51 +/- 17, and 137 +/- 75 mg/dL, respectively. The proportion of patients on statin treatment was 49.7%, and those with body mass indices between 25 and 30, 30 and 35, and >35 kg/m(2) were 35%, 15%, and 4%. We observed a high prevalence of chronic kidney disease, hypertension, dyslipidemia, and obesity among renal transplant patients. Suboptimal control was frequent and control of some of these complications was far below targets established for nontransplant patients despite progressive intensification of therapy with functional graft decline. The findings of this study may have an impact on the management of renal transplant recipients. PMID- 19545699 TI - Low levels of Th1-type cytokines and increased levels of Th2-type cytokines in kidney transplant recipients with active cytomegalovirus infection. AB - BACKGROUND: Cytomegalovirus (CMV) infection is a major complication after kidney transplantation. It is clear that Th1 and Th2 cell subsets are of major importance in determining the class of immunoprotective function in infectious diseases. Given the strong influence exerted by Th1- and Th2-type immunity on the outcome of infections, we felt it important to elucidate the levels of Th1- and Th2-type cytokines to CMV-related antigens in kidney recipients and to identify antigens that play an essential role in preventing the development of CMV infection and/or disease. METHODS: One hundred twenty subjects were followed for CMV infection by the antigenemia assay. We investigated peripheral blood mononuclear cells (PBMCs) responses to five CMV-related peptide antigens (pp65, gB, pp150, pp28, and pp38). Stimulation index was determined by radioactive thymidine uptake, while the production of Th1-type cytokines (interferon-gamma and tumor necrosis factor-alpha) and Th2-type cytokines (interleukins-4 and -10) were measured by enzyme-linked immunosorbent assay. RESULTS: The levels of Th1 type cytokine production after stimulating PBMCs with CMV-related antigens gB and pp150 resulted in significant decreases in the levels of interferon-gamma, while pp65, pp150, and pp38 produced significant decreases in the level of tumor necrosis factor-alpha between the two groups (P < .05). For Th2-type cytokines only pp28 produced a significant increase in the level of interleukin-10 between the two groups (P < .05). Regarding the Th1:Th2 ratios, a lower Th1-bias was observed among the CMV-positive patients for PBMCs stimulated with three CMV related antigens (pp65, pp38, and pp28). CONCLUSION: Low levels of Th1-type cytokines and increased levels of Th2-type cytokines upon stimulation with CMV related peptide antigens were associated with reduced cell-mediated immunity to CMV, thus seeming to correlate with active CMV infections. PMID- 19545698 TI - Prognostic effect of lymphocyte subgroup CD4+ and CD8+ cells in peripheral blood in renal transplant patients with cytomegalovirus viremia. AB - OBJECTIVE: Our aim was to evaluate the prognostic effect of peripheral blood lymphocyte subgroup CD4+ and CD8+ cells on renal transplant patients with cytomegalovirus (CMV) viremia. MATERIALS AND METHODS: Using 41 renal transplant patients with CMV-PCR(+) in peripheral blood and stable values of serum creatinine (SCr), we evaluated the changes in lymphocyte subgroup CD4+ and CD8+ cells with onset of antiviral therapy with gancyclovir for treatment of pneumonia. We compared patients with or without pneumonia. RESULTS: The lower the peripheral blood lymphocyte subgroup CD4+ and CD8+ cell numbers, the higher the incidence of CMV pneumonia. The numbers of CD4+ and CD8+ cells at 1 month posttransplantation and at the time of CMV-PCR(+) detection were significantly lower than those before transplantation in the CMV pneumonia group (P < .01) and also in the nonpneumonia group. CONCLUSIONS: The decrease in peripheral blood lymphocyte subgroup CD4+ and CD8+ cells after renal transplantation in patients with CMV viremia showed prognostic value for pneumonia. Increased CD4+ and CD8+ cells in peripheral blood combined with preemptive therapy may reduce the incidence of pneumonia among patients with CMV viremia. PMID- 19545700 TI - Severe bone marrow failure due to valganciclovir overdose after renal transplantation from cadaveric donors: four consecutive cases. AB - Valganciclovir is an l-valyl ester pro-drug of ganciclovir that was initially used to treat cytomegalovirus (CMV)-associated retinitis in patients with human immunodeficiency virus. Currently, it is also indicated for the prevention of CMV disease in solid-organ transplantation. It is primarily eliminated via the kidneys through glomerular filtration and tubular secretion. Decreased renal function results in decreased drug clearance. Valganciclovir has been reported to cause usually mild to moderate hematologic adverse effects such as leukopenia, neutropenia, anemia, thrombocytopenia, and pancytopenia. Severe and fatal bone marrow depression has been described in 1 adult patient. Herein, we describe the cases of 4 patients with end-stage renal disease who underwent cadaveric renal transplantation and received valganciclovir prophylaxis for CMV at a standard dose of 900 mg/d despite persistant renal failure. This therapy resulted in severe bone marrow failure after 18 to 20 days in all 4 patients, with fatal infections in 2 patients. This report demonstrates the in vivo pharmacodynamics of valganciclovir overdose in terms of hematotoxicity in the setting of renal impairment. Valganciclovir, as its derivative ganciclovir, should be used cautiously in patients with renal impairment. PMID- 19545701 TI - Pulmonary fungal infections in kidney transplant recipients: an 8-year study. AB - Invasive fungal infections are among the most important causes of mortality among transplant patients. One of the most common manifestations of these infections is pulmonary fungal infection (PFI). The present study sought to evaluate the rate of PFI in kidney transplant patients. MATERIALS AND METHODS: We retrospectively analyzed the data of 595 patients who underwent kidney transplantation from February 1999 to February 2007. Bronchoalveolar lavage (BAL) culture and tissue biopsy were used to confirm PFI. RESULTS: Thirteen of 595 patients (2.2%) experienced PFI. The most common pathogen (8/13, 41.5%) was Aspergillus, with 5 (38.5%) infected with Aspergillus only, 2 (15.4%) with both Aspergillus and Candida, and 1 (7.7%) with Aspergillus and mucormycosis. Seven of 13 (53.8%) died and 4 (30.7%) lost the transplanted kidney. Immunosuppressive therapy following rejection and prescription of broad spectrum antibiotics were the most important risk factors for fungal infections in these patients. CONCLUSIONS: Fungal infections are among the most important causes of mortality among transplant patients, of which the most common manifestation is pulmonary. Immunosuppressive therapy and broad spectrum antibiotics are important risk factors, and Aspergillus is the most common pathogen responsible for fungal infections. PMID- 19545703 TI - Kidney graft function in long-term cyclosporine and tacrolimus treatment: comparative study with nephrotoxicity markers. AB - Calcineurin inhibitors improve kidney allograft survival in the posttransplantation period; however, they may cause nephrotoxicity. The objective of this study was to compare the effect of cyclosporine (CsA) and tacrolimus (Tac) treatment on the transplanted kidney. The study included 219 patients aged 21 to 65 years. Of these, 120 (39 women and 81 men) were treated with CsA and 99 (38 women and 61 men) were treated with Tac. Patients were divided into 4 groups depending on the time since kidney transplantation. We evaluated urine markers of nephrotoxicity: proximal tubular cells lysosomal enzymes (N-acetyl-beta-d glucosaminidase [NAG] and its isoform NAG-B, beta-d-galactosidase, and beta glucouronidase) and brush border enzymes (alanyl aminopeptidase and gamma glutamyl transpeptidase). Urine activities of nephrotoxicity markers were compared in CsA- and Tac-treated patients groups depending on the duration of treatment and allograft function as measured by serum creatinine concentration. Correlation studies between CsA and Tac levels and enzyme activities were performed in both groups and in the entire patient cohort. NAG and its isoform NAG-B seemed to be the most reliable markers of nephrotoxicity. Despite the significant correlation between NAG urine activity and serum creatinine concentration in the CsA group, there were no significant differences in NAG or NAG-B activities between CsA- and Tac-treated graft recipients. PMID- 19545702 TI - Enteric coating of mycophenolate reduces dosage adjustments. AB - Mycophenolate mofetil (MMF) and enteric-coated mycophenolate sodium (EC-MPS) are bioequivalent. However, the effectiveness of MMF may be limited by gastrointestinal (GI) side effects. This study assessed the relationship between the number of medication dosage adjustments and posttransplantation side effects. In a review of 109 kidney transplant patients, 65 initially received MMF and 44 initially received EC-MPS. The incidences of patient-reported GI complications were significantly different: MMF 45.5% vs EC-MPS 35.3% (P = .0194). The proportions of patients requiring dosage adjustment due to GI complications were MMF 5.9% and EC-MPS 2.3% (P < .0001). Patients receiving MMF were more likely to experience GI complications resulting in dosage adjustment (odds ratio = 9.9; P = .0306). The incidences of acute rejection, cytomegalovirus (CMV), and leukopenia resulting in dosage adjustment were not significantly different. Patients receiving MMF required more immunosuppressive medication adjustments, which may complicate care and decrease overall compliance. PMID- 19545704 TI - Long-term follow-up of 100 high-risk renal transplant recipients converted from calcineurin inhibitors to sirolimus: a single center experience. AB - While conversion of stable renal transplant recipients (RTR) from calcineurin inhibitors (CNI) to sirolimus (SRL) is safe and effective, it is still under investigation for recent, high-risk cases. We studied the long-term effects of conversion of high-risk subjects maintained on a CNI, mycophenolate mofetil, plus steroid regimen to SRL, mycophenolate mofetil, plus steroid on graft and patient outcomes. We retrospectively reviewed the first 100 RTR converted to SRL treatment over approximately 5 years. The main indications for conversion were biopsy-proven acute rejection (BPAR), CNI toxicity, CNI elimination, and acute tubular necrosis (ATN). Exclusion criteria were limited to bone marrow suppression. The overall mean +/- SD age was 38.5 +/- 15.6 years, including pediatric and geriatric age groups. Mean +/- SD body mass index (BMI) was 28.99 +/- 8.0 and 40% had a BMI > 30. There were 40% RTR from deceased donors and 60% showed 4 to 6 HLA mismatches. Preconversion total BPAR and steroid-resistant rejection incidences were 35% and 14%, respectively. Mean +/- SD time to start of SRL was 11.9 +/- 22.8 months posttransplantation. Proteinuria > 2 g/d, leukopenia, and hyperlipidemia increased significantly after conversion (P = .001, P = .0003, and P = .0001, respectively). Patient and graft survivals were 95% and 90%, respectively. There was significant improvement in graft function postconversion (P < .0001). There was a high incidence of side effects and cases of SRL discontinuation. Multivariate analysis demonstrated the influence of bone marrow suppression, obesity, hyperlipidemia, nutritional status, proteinuria, and graft function on graft and patient outcomes. We concluded that conversion from CNI to SRL was effective among high-risk RTR, but with a high incidence of adverse events during long-term follow-up. PMID- 19545705 TI - A 6-month, multicenter, single-arm pilot study to evaluate the efficacy and safety of generic tacrolimus (TacroBell) after primary renal transplantation. AB - OBJECTIVE: Tacrolimus has been shown to be an important immunosuppressive agent in organ and bone marrow transplantation. Previously, we reported that there were no statistically significant differences between the pharmacokinetic parameters of the oral formulation of generic tacrolimus (TacroBell) and the conventional formulation (Prograf). This study was designed to evaluate the efficacy and safety of oral capsules of TacroBell in de novo renal transplantation. METHODS: Ninety-six renal transplant recipients from 9 transplantation centers in South Korea were enrolled between November 2005 and July 2007. De novo renal recipients ranged from 19-65 years old. Ninety-four patients who underwent renal transplantation were administered study drug at least one time in the intent-to treat (ITT) analysis. This phase 4 clinical trial was a 26-week, open-label, noncomparative, multicenter study. RESULTS: An acute rejection episode developed in 10/94 recipients (10.6%, 95% confidence interval, 4.4%-16.9%). There were no patient deaths during the study. The 6-month graft survival rate was 96.8%. CONCLUSION: Based on this study, treatment with TacroBell is considered to be efficient and safe after primary renal transplantation. PMID- 19545706 TI - Use of bioelectrical impedance analysis to assess liver steatosis. AB - BACKGROUND: The shortage of donor livers has led to increased utilization of steatotic marginal livers. Bioelectrical impedance analysis (BIA) uses the principles of electric current flows through tissue, with less resistance offered if the water content is high and the opposite in the presence of fat. Our hypothesis was that liver steatosis would result in an increased resistance to current flow, and correlate with the degree of liver steatosis. METHODS: Before studying cadaveric donor livers for transplantation, this study was performed in patients undergoing liver resection. A total of 37 patients undergoing liver resection for cancer were analysed with BIA, using a handheld, specially calibrated Maltron BIA analyser (BioScan 915) with modified tertrapolar electrodes. These electrodes were applied to the liver surface and resistance was recorded. To validate the results of BIA, a liver biopsy was performed. Histopathology was graded quantitatively as no steatosis, mild, moderate, or severe steatosis according the percentage of fat as well as qualitatively by type of fat (micro and macrovesicular). RESULTS: Bioelectric resistance showed a correlation with macroveiscular steatosis (P = .03). CONCLUSION: BIA is a simple, noninvasive technique and its use should be explored in donor livers to assess steatosis. PMID- 19545707 TI - A model of risk and protective factors influencing the postoperative course of living liver donors. AB - Living donor liver transplantation is increasing as an effort to overcome the organ shortage for patients with terminal liver diseases. To maintain high medical and ethical standards, donors are evaluated before and after donation to assess their suitability, monitor their postoperative courses, and minimize risks. A psychological/psychosocial evaluation is included in the assessment at most transplantation centers. Due to the high risk and the challenging process of recovery and integration of the experience into one's life, it is important to identify donors with suitable psychological/psychosocial profiles. The psychosocial evaluation is performed on the basis of medical ethical standards, common sense, professional experience, and only a few systematic observations. Some studies have identified factors, such as complications for the recipient, as a possible influence on the donor's postoperative course, while others have sought to link donor outcomes with scores on psychometric instruments prior to donation. We suggest that it is not one, but the interplay of several factors that affects the donor's postoperative course, including decision autonomy and embedding the donation into a meaningful context, in addition to coping skills and recipient outcome. Based on pilot data, clinical observations, and a study of the literature, we developed a model of risk and protective factors influencing the donors' postoperative courses, which could be used to assess living liver donors psychosocially before and after donation, thus enhancing selection and support. PMID- 19545708 TI - Preservation of recipient middle hepatic vein for drainage of anterior sector veins in adult-to-adult living-donor liver transplantation. AB - The inclusion of donor middle hepatic vein (MHV) in right-lobe living-donor grafts and the need for reconstruction of the MHV tributaries have long been controversial areas in living-donor liver transplantation. We report technical details in restoration of venous drainage of the anterior sector (segments V and VIII) of the right lobe of the liver graft using a preserved MHV from the recipient liver, and address the issue of reconstruction of donor MHV tributaries without use of an interposition graft. We review clinical situations in which restoration of outflow drainage of the anterior segment of the liver graft should be considered. PMID- 19545709 TI - Hepatic resection for hepatocellular carcinoma meeting Milan criteria in Child Turcotte-Pugh class a patients with cirrhosis. AB - This study evaluated whether hepatic resection is a reasonable strategy as an initial treatment for hepatocellular carcinoma (HCC) meeting Milan criteria in patients with compensated cirrhosis. From the database of 435 consecutive patients with resection of HCC between July 1994 and May 2007, 213 patients were found to have Child-Turcotte-Pugh class A cirrhosis and HCC meeting Milan criteria, as shown by preoperative image studies. We examined long-term survivals and patterns of recurrence after hepatic resection among those patients. Overall survival rates at 1, 3, 5, and 10 years were 92%, 78%, 69%, and 52%, respectively, and 1-, 3-, 5-, and 10-year disease-free survival rates were 79%, 57%, 44%, and 19%, respectively. Pathological review indicated that 36/213 patients (16.9%) had another nodule and/or gross vascular invasion. Microvascular invasion, tumor size, and histological grade of cirrhosis were independent risk factors for recurrence. Sixty percent of recurrent cases met the Milan criteria. The six patients who underwent living donor salvage liver transplantation (OLT) for intrahepatic recurrence were alive without recurrence at a median of 24 (range = 8-31) months. These favorable data suggest that hepatic resection is a good option for small HCCs in patients with compensated cirrhosis; and salvage OLT may be reserved for patients with recurrences. PMID- 19545710 TI - Initial clinical effect of intraportal insulin administration on liver graft regeneration in adult patients underwent living donor right lobe liver transplantation. AB - OBJECTIVE: Insulin is one factor responsible for hepatotrophic regeneration in animal models. This study assessed the clinical effects of intraportal administration of insulin on liver graft regeneration in adult patients undergoing right lobe living donor liver transplantation (LDLT). METHODS: Between July 2005 and September 2007, 19 right lobe LDLT adult recipients voluntarily received posttransplant intraportal insulin administration. The present study describes 15 patients without postoperative vascular and bile duct complications, with more than 1 month survival and with complete clinical data who were enrolled to receive intraportal insulin therapy (group I; n = 15). Another consecutive 15 right lobe LDLT adult recipients without any stimulation regeneration who met the same criteria were enrolled in as noninsulin therapy control group (group NI; n = 15). Group I recipients were treated postoperatively with intraportal insulin infusion, as follows. An 18-gauge catheter was inserted into right gastro-omental vein during surgery, to administer regular insulin just after the operation at the rate of 2 U/h for 1 week. Graft volume (GV) was measured by computed tomography on postoperative days (POD) 7 and 30. Liver functions and serum insulin levels were also measured at POD 7 and POD 30. The liver graft regeneration rate was defined as ratio of posttransplant GV/harvested GV and posttransplant graft-to-recipient weight ratio (GRWR)/operative GRWR. RESULTS: The rate defined as ratio of POD 7 GV/harvested GV among group I was significantly greater than that of group NI (186.07 +/- 35.40% vs 160.61 +/- 22.11%; P < .05). The rate defined as ratio of POD 7 GRWR/operation GRWR was also significantly higher in group I than group NI (178.95 +/- 35.84% vs 156.56 +/- 18.53%; P < .05), whereas there was no significant difference in terms of regeneration rates at 1 month post-LDLT. Intraportal insulin administration may significantly downregulate POD 7 total bilirubin, aspartate aminotransferase, and alanine aminotransferase levels (P < .05). These results suggested that intraportal insulin administration augmented liver regeneration during the first postoperative week by improving hepatic function in LDLT recipients. PMID- 19545711 TI - Using the Clavien grading system to classify the complications of right hepatectomy in living donors. AB - INTRODUCTION: The ratios of complications for living related liver donors after right hepatectomy differ widely among numerous single institutions. This study sought to use the Clavien classification system to define and graded the severity of these complications. MATERIALS AND METHODS: This study retrospectively analyzed the outcomes of 160 consecutive living donor right hepatectomies performed between July 2002 and February 2008. Complications among living donors for liver transplantation after right hepatectomy were stratified according to the Clavien classification of postoperative surgical complications. RESULTS: Fifty-two living donors displayed one or more perioperative complications Grade 1 complications were recorded in 18.1%; grade 2 in 6.3%; grade 3a in 5%; and grade 3b in 3.1%. Biliary complications were the most frequent. No donor mortality was present in this series. CONCLUSIONS: The Clavien grading system is useful to comparise surgical outcomes. This study demonstrated that donor right hepatectomy was a relatively safe procedure, but reducing donor complications after right hepatectomy has to be the first priority during the entire process of living related transplantation. PMID- 19545713 TI - Outcome of combined liver and kidney transplantation in hepatitis C: a single center long-term follow-up experience. AB - INTRODUCTION: Hepatitis C (HCV) cirrhosis is the prevalent liver disease requiring liver transplantation in the United States. Candidates who also have end-stage renal disease, chronic renal disease stage 4, or prolonged hepatorenal syndrome are considered for combined liver and kidney transplantation (CLKT). MATERIALS AND METHODS: We performed a retrospective study of HCV(+) and HCV(-) CLKT patients with more than 12 months of follow-up and HCV(+) patients with isolated liver transplant (OLT) to compare the outcomes of various groups. RESULTS: Since 1988, 2983 OLTs were performed at our institution including 58 CLKTs. Of these, 23 were HCV(+) subjects who were significantly older than HCV(-) CLKT patients. Race, pretransplant dialysis time, renal indication for CLKT, Model for End-stage Liver Disease score, donor age, liver and kidney rejection as well as occurrence of posttransplant hypertension were similar among HCV(+) and HCV(-) CLKT patients. Posttransplant diabetes was observed in 80% of the HCV(+) group and 30% of the HCV(-) group (P = .01). Renal function seemed to be better in HCV(-) when compared with HCV(+) subjects at 5 years (P = .09). Overall patient survival for HCV(+) CLKT, HCV(-) CLKT, and HCV(+) OLT groups at 1, 2, and 5 years were not significantly different (P = .6). CONCLUSION: HCV positivity should not exclude appropriate candidates for CLKT. PMID- 19545712 TI - Liver transplantation for primary biliary cirrhosis: results of aggressive corticosteroid withdrawal. AB - INTRODUCTION: A subset of patients with primary biliary cirrhosis (PBC) may require long-term corticosteroid (CS) therapy following liver transplantation (OLT) due to concern over the possibility of recurrence. Our center has attempted to minimize CS use in all of our OLT recipients. In this study, we review our experience in this cohort to determine (1) patient outcome including PBC recurrence following transplantation and (2) the long-term requirement for CS use in PBC patients. METHODS: From 1988 to 2006, 1102 OLTs were performed in 1032 adults at the University of Colorado, of which 70 patients (6.8%) with PBC received 74 allografts. Bivariate and multivariate analyses were used to evaluate predictors of CS withdrawal. Thirteen potential predictors of CS discontinuation were considered: age, gender, body mass index (BMI), race, type of graft (cadaveric or living donor [LD]), recurrence of PBC, warm ischemia time, and immunosuppressant. RESULTS: Overall survival at 5 years was 85%. The 1-, 5-, and 10-year recurrence-free survivals were 90%, 72%, and 54%, respectively. PBC recurred in 18 patients (25.7%). Of these, none received a second transplant due to disease recurrence. At the time of last follow-up, 73% of recipients were steroid free. Independent predictors of CS discontinuation are age (>54; P = .0059) and LD graft type (P = .0008). Conversely, cyclosporine (P = .0007), female gender (P = .0216), and BMI > 31 (P = .0306) were negatively associated with CS withdraw. Importantly, steroid discontinuation did not influence PBC recurrence. CONCLUSIONS: While long-term outcomes in PBC patients are favorable, disease recurrence can generally be managed medically without the need for a second transplant. Using an aggressive CS minimization approach, nearly three quarters of the patients were CS free at the time of last follow-up. Increasing age and LD grafts were associated with successful CS withdraw. Conversely, cyclosporine use, female gender, and increasing BMI were associated with unsuccessful steroid discontinuation. PMID- 19545714 TI - Liver transplantation for hepatitis B virus-related hepatocellular carcinoma: one center's experience in China. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is the fifth most common and the third most deadly cancer worldwide, with more than half a million identified cases and about a similar number of subjects succumb to it each year. This study sought to evaluate our results of liver transplantation for HCC to identify prognostic factors. METHODS: Between December 2001 and December 2006, 224 patients (205 men, 19 women; age range, 15-75 years) with HCC underwent orthotopic liver transplantation (OLT) at our center. All grafts were from deceased donors. There were 68 cases within Milan criteria (30.3%), 32 cases beyond Milan criteria but within UCSF (University of California, San Francisco) criteria (14.3%), and 124 cases beyond UCSF criteria (55.4%). RESULTS: The overall 1-, 3-, and 5-year patient cumulative survival rates were 82.5%, 60.1%, and 51.5%, respectively. The survival rates were comparable between patients within Milan and UCSF criteria, but were significantly greater than that of patients beyond UCSF criteria. Multivariate analysis revealed alpha fetoprotein (AFP) >or= 800 microg/L, vascular invasion, and poor tumor differentiation to be independent prognostic factors. CONCLUSION: OLT is a safe and effective treatment for hepatitis B virus related HCC. Compared with Milan criteria, UCSF criteria successfully expanded the indication without deteriorating the prognosis significantly, while preoperative AFP >or= 800 microg/L, vascular invasion, and poor tumor differentiation indicated poor survival. PMID- 19545715 TI - Effects of cardiac valve dysfunction on perioperative management of liver transplantation. AB - OBJECTIVE: Our aim was to investigate the effects of cardiac valve dysfunction on perioperative management of orthotopic liver transplantation (OLT) among a retrospective cohort. PATIENTS AND METHODS: Three hundred forty-six patients underwent echocardiographic (ECHO) examination prior to OLT. Data of patients with valvular dysfunctions were compared to subjects with normal ECHO. We evaluated patient characteristics, operation variables, hemodynamic course, blood products, fluid and drug requirements, extubation, and mortality rates. RESULTS: Ninety-five patients (27.5%) with cardiac valve dysfunction were classified as mitral valve insufficiency (MVI; n = 32), tricuspid valve insufficiency (TVI; n = 23), or both MVI and TVI (n = 40). One hundred fifty-two patients displayed normal ECHO examinations (control). Ninety-nine patients with other pathologies were excluded from the study. Systemic vascular resistance was significantly lower among the MVI group, and cardiac output (CO) significantly higher in the MVI and both MVI and TVI groups compared with controls. More MVI and both MVI and TVI patients required epinephrine compared with controls. The number of patients who required blood transfusion was higher in the MVI than the control group (P < .05). Patient characteristics, end-stage liver failure scores, duration of operations, hemodynamic variables, incidence of postreperfusion syndrome, mean doses of ephedrine and epinephrine, red blood cells, fresh frozen plasma and fluid requirements, number of patients extubated immediately after surgery, and mortality rates were not different between the groups. CONCLUSIONS: Our study demonstrated that cardiac valve dysfunction may be associated with end-stage liver disease among patients undergoing OLT. Patients with MVI or both MVI and TVI required more care in perioperative management. PMID- 19545716 TI - An early increase in gamma glutamyltranspeptidase and low aspartate aminotransferase peak values are associated with superior outcomes after orthotopic liver transplantation. AB - BACKGROUND: Prediction of prognosis after liver transplantation (OLT) remains difficult. The present study determines if standard laboratory parameters measured within the first week after OLT correlate with outcome. PATIENTS AND METHODS: Laboratory parameters measured within the first weak after OLT of 328 patients were grouped either graft loss or death within 90 days after (group 1: graft loss; group 2: death; group 3: neither graft loss nor death within 90 days). RESULTS: Peak AST and ALT were significantly lower in group 3 (1867 and 1252 U/L) than in group 1 (4474 and 2077 U/L) or 2 (3121 and 1865 U/L). Bilirubin was significantly lower and gamma-GT significantly higher in group 3 compared to groups 1 and 2. In multivariate analysis, high AST peaks were independently associated with death or graft loss within 90 days. An increase in gamma-GT and low bilirubin early after transplantation were found to be independently associated with superior outcome. DISCUSSION: Unexpectedly, a disproportionate rise in gamma-GT was associated with graft and patient survival of more than 90 days. This might be explained by regeneration phenomena in the liver indicative of a well functioning graft. PMID- 19545717 TI - Adherence correlates in liver transplant candidates. AB - The study sought to investigate the psychiatric and psychosocial correlates of multidimensional adherence among liver transplant candidates. A liver transplant candidate sample (N = 100), comprising consecutive patients attending outpatient clinics of a liver transplantation central unit, was assessed by means of the Multidimensional Adherence Questionnaire (MAQ; Telles-Correia 2007), the Diagnostic and Statistical Manual of Mental Disorders, Revised 4th Edition (DSM IV-TR) criteria, the Toronto Alexithymia Scale (TAS), the NEO Five-Factor Inventory (NEO-FFI) personality inventory, the Hospital Anxiety and Depression Scale (HADS), the Brief COPE, and the Psychological Assessment of Candidates for Transplantation (PACT). We found that multidimensional adherence positively correlated with personality traits (agreeableness), good social support, and coping strategies (planning). PMID- 19545718 TI - Menstrual function in female liver transplant recipients of reproductive age. AB - BACKGROUND AND AIM: End-stage liver failure is associated with severe abnormalities in menstrual and reproductive function. These abnormalities may be reversed by successful orthotopic liver transplantation (OLT). The aim of the study was to investigate menstrual patterns and sex hormone profiles among female liver transplant recipients of reproductive age. METHODS: The study group consisted of 24 women of reproductive age with end-stage liver failure who underwent successful OLT. Menstrual patterns and sex hormone profiles were analyzed before as well as 3 and 12 months after OLT. Twenty-seven healthy women of reproductive age served as controls. Biochemical parameters of liver function were assessed before and after OLT. RESULTS: Amenorrhea was the most commonly observed abnormality of menstrual cycle in women with end-stage liver failure (71% of patients). The recurrence of regular menstrual cycles was observed in 35% of patients 3 months after OLT. The percentage increased to 70% at 1 year after grafting and was clearly associated with stabilization of liver function. Similar levels of follicle stimulation hormone (FSH), luteinizing hormone (LH), prolactine (PRL), and testosterone (T) as well as lower levels of estradiol (E(2)), dehydroepiandrostendione sulphate (DHEA-S), and progesterone, (P) were observed in patients with liver failure compared with healthy women. We observed normalization of E(2) and DHEA-S levels after OLT. CONCLUSIONS: Amenorrhea, the most common menstrual disturbance in women with end-stage liver failure, may be reversed by OLT. One year after OLT menstrual bleedings were noted in 74% of patients of reproductive age. The recurrence of reproductive function indicated the need for effective and safe family planning methods in that group of patients. PMID- 19545719 TI - Orthotopic liver transplantation for hepatocellular carcinoma: one center's experience in the Northeast of Brazil. AB - Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world and the third leading cause of cancer-related death. In this study, we sought to assess the outcome of patients with HCC who underwent orthotopic liver transplantation (OLT) in a center in the northeast of Brazil. Between May 2002 and July 2008, 294 OLTs were performed at our center. In 45 patients, HCC was confirmed by histological examination of the explant. Patients were predominantly men of ages ranging from 14-67 years. Hepatitis C virus was involved in 55.4% of the cases. Alpha fetoprotein (AFP) levels were normal in 65.2% of the patients and surpassed 100 ng/mL in only 10.4%. The median waiting time on the list was 10 months. Seventeen patients (37.7%) presented a solitary nodule, 19 (42.2%) had 2 or 3 nodules, and 9 patients (20%) had more than 3 nodules. The maximal diameter of the largest tumor was <3 cm in 26 patients (57.7%) and exceeded 5 cm in 6 patients (13.3%). Ten tumors were well differentiated, 32 were moderately differentiated, and 3 were poorly differentiated. Eleven tumors showed microvascular invasion. There have been 4 tumor recurrences. There was an association between microvascular invasion and tumor recurrence with a statistically significant relative risk. In conclusion, OLT is an excellent option for patients with HCC. The recurrence rate was low (<10%). However, we believe that more prospective studies are needed about OLT beyond the Milan criteria because our study suggested that microvascular invasion may be more important than tumor size or number. PMID- 19545720 TI - Anxiety and depression levels of patients undergoing liver transplantation and their need for training. AB - The purpose of this research was to determine the levels of anxiety and depression and the need for training of patients who had undergone liver transplantation. This descriptive research included patients who had undergone liver transplantation at least 1 month prior and had follow-up between June and November 2006. The sample consisted of 64 patients who completed a questionnaire consisting of 14 semistructured questions to determine levels of anxiety and depression using the Hospital Anxiety and Depression Scale (HADS) developed by Zigmond and Snaith (Acta Psychiatr Scan 67:361, 1983). Paired Student t test was used to assess the need for patient training and to compare the mean scores of the scale Chi-square tests were used for data. Eighty-nine point five percent of anxious patients depressed; numerical 44.4% of those who were not anxietic were depressed. To determine the need for patient training, issues that are worrisome/aggrieving for patients were asked, but a priority of issues was not associated with depression and anxiety of patients. The scale was more sensitive to depression than to anxiety. I will be more appropriate to professionally evaluate anxiety and depression and consider the need for patient training. PMID- 19545721 TI - Duct-to-duct biliary reconstructions and complications in 100 living donor liver transplantations. AB - OBJECTIVE: We evaluated the risk factors for biliary complications and surgical procedures for duct-to-duct reconstructions in adult living donor liver transplantation (LDLT). PATIENTS AND METHODS: From February 2005 to March 2008, we performed 100 cases of adult LDLT with duct-to-duct biliary reconstruction, using 64 right lobe grafts, 33 left lobe grafts, and 3 right lateral grafts. We employed 4 types of duct-to-duct procedures: all interrupted 6-0 Prolene suture (group 1, n = 9); continuous posterior and interrupted anterior wall 6-0 Prolene suture (group 2, n = 49); all continuous 7-0 Prolene suture (group 3, n = 26); and all continuous 7-0 Prolene suture with external stent (group 4, n = 16). Biliary complications were defined as an anastomosis stricture or a leakage. RESULTS: Thirty-four patients experienced biliary complications during the follow up period (median, 27 months). The incidence of stricture was 27% and that of leakage, 8%. There were no perioperative, intraoperative, or anatomic risk factors for biliary complications, except the type of duct-to-duct procedure. Group 1 and 2 patients showed higher incidences of biliary strictures than groups 3 and 4 (43.1% vs 4.7%; P = .00). Group 3 patients experienced a higher incidence of bile leakage than the other groups (23.1% vs 2.7%; P = .004). CONCLUSIONS: The type of biliary reconstruction is a factor affecting biliary complications following duct-to-duct anastomosis in LDLT. Duct-to-duct biliary anastomosis with 7-0 monofilament suture and a small external stent is a feasible procedure in LDLT that significantly reduces the incidence of biliary complications. PMID- 19545722 TI - Diabetes mellitus after living donor liver transplantation: data from mainland China. AB - Most reported data on posttransplantation diabetes mellitus (PTDM) are from Western countries with patients who underwent deceased donor liver transplantation. A retrospective study was performed to assess the prevalence and predictive factors of PTDM in the context of living donor liver transplantation (LDLT) in the Chinese population using the definition of PTDM proposed in 2003 by the World Health Organization and the American Diabetes Association. The prevalence of DM after LDLT in our study was 25% (21/84), and the incidence of PTDM was 14.9% (11/74) with 64% of cases diagnosed within 3 months after LDLT; 9.5% were observed to show impaired fasting glucose postoperatively. Multivariate analysis identified body mass index >or= 25 kg/m(2) before LDLT as the only independent risk factor for developing PTDM. Only one patient was operated for hepatitis C virus (HCV) infection. Hepatitis B virus (HBV)-related diseases were common in our study population, accounting for 78.6% of all patients. Both HCV and HBV infection status were not independent risk factors for developing PTDM. In addition, a greater tacrolimus trough blood level in the PTDM group versus no DM group was observed at 3 months post-LDLT (11.03 ng/mL vs 4.87 ng/mL). The mean tacrolimus dose was not significantly different between the two groups. In conclusion, PTDM was prevalent among Chinese LDLT recipients. PMID- 19545724 TI - Change of peripheral blood mononuclear cells IFN-gamma, IL-10, and TGF-beta1 mRNA expression levels with active human cytomegalovirus infection in orthotopic liver transplantation. AB - AIM: To analyze the expression levels of interferon gamma (IFN-gamma), interleukin 10 (IL-10), and transforming growth factor beta 1 (TGF-beta1) mRNA in peripheral blood mononuclear cells (PBMCs) in liver transplanted recipients with active HCMV infection. METHODS: PBMCs were isolated from 20 liver transplanted recipients with active HCMV infection and 20 recipients without HCMV infection. The expression levels of IFN-gamma, IL-10 and TGF-beta1 mRNA in PBMCs were measured by TaqMan real-time reverse transcriptase-polymerase chain reaction (RT PCR). The results were compared with that from 20 healthy individuals. RESULTS: The expression level of TGF-beta1 mRNA was significantly increased in the active HCMV infection group compared with that in stable group or healthy group (P < .001). The expression level of IL-10 mRNA was significantly increased in the active HCMV infection group compared with the healthy group (P = .001). However, the IFN-gamma mRNA expression level was significantly decreased in the active HCMV infection group compared with that in the stable group or the healthy group (P < .05). CONCLUSION: Cytokine production plays a role in the HCMV infection. This may provide an important clue to a better understanding of the pathogenesis in liver transplanted recipients with active HCMV infection. PMID- 19545723 TI - Elevated CD4+/CD25+ T-cell frequency and function during hepatitis C virus recurrence after liver transplantation. AB - BACKGROUND/AIM: Factors involved in hepatitis C virus (HCV) recurrence versus acute cellular rejection are not fully understood. The aim of the present study was to investigate whether patients with recurrence after liver transplantation (OLT) showed similar CD4(+)/CD25(+) cell frequency and function as those who became chronically infected. PATIENTS AND METHODS: After written informed consent, we enrolled 20 patients (group A) who underwent OLT with HCV recurrence within 6 months. HCV-RNA and hypertransaminasemia were used to assess the reactivation of viral hepatitis. CD4(+)/CD25(+) T cells were enumerated using a flow cytometry assay, gated on CD3 cells, stained for FoxP3. After immunomagnetic sorting (Dynal, Oslo, NW), Treg suppressor activity was measured, as the ability to inhibit proliferation of autologous CD4(+)/CD25(-) T cells (anti-CD3/CD28 stimulation-1:2, 1:20 ratio). Eight patients with acute hepatitis C who evolved to a chronic infection after 6 months (group B) were used as positive controls, while 10 healthy individuals were negative controls (group C). RESULTS: We did not observe any difference in CD4(+)/CD25(+) frequency or function among group A compared with group B (CD4(+)/CD25(+) = 14% +/- 2% versus CD4(+)/CD25(+) = 16% +/ 3%), although both groups were significantly increased with respect to group A (CD4(+)/CD25(+) = 6% +/- 3%; Mann-Whitney U test, P < .01). CONCLUSION: Patients developing HCV recurrence after OLT have the same immunoregulatory network as patients with acute hepatitis C evolving to persistent infection, likely suggesting that CD4(+)/CD25(+) numbers may be a marker to predict recurrence of HCV after OLT. PMID- 19545725 TI - Asynchronous kidney allograft loss after simultaneous pancreas-kidney transplantation: impact on pancreas allograft outcome at a single center. AB - OBJECTIVE: We analyzed the clinical evolution of pancreas allografts in simultaneous pancreas-kidney transplantation (SPKT) cases after asynchronous kidney allograft loss and kidney retransplantation at a single non-United States center. PATIENTS AND METHODS: We performed a retrospective analysis of 168 SPKT from December 2000 to June 2007. RESULTS: The 5-year kidney allograft survival rate was 71%. Excluding cases of death with a functioning graft after SPKT (n = 35; 74.4%), 12 kidney allografts were lost due to acute rejection (n = 7; 15%) or chronic allograft nephropathy (n = 5; 10.6%). Delayed graft function contributed to kidney allograft loss. Five of 12 patients underwent kidney retransplantation. Sixty percent of pancreas allografts were lost after this procedure, which was attributed to either the diabetogenic effects of the immunosuppressive regimen or to the perioperative stress. Oral glucose tolerance tests performed before kidney retransplantation identified patients with good pancreas allograft function versus those with intolerance on glucose tests who received reduced glucocorticoid doses. CONCLUSIONS: In SPKT, pancreas allograft function was seriously affected by kidney retransplantation. Oral glucose tolerance tests performed before kidney retransplantation were helpful to assess beta-cell function and suggest prescription of lower steroid doses to decrease the pancreas allograft dysfunction. PMID- 19545726 TI - Staged enteric conversion after duodenal necrosis in simultaneous kidney and pancreas transplant from a donor after cardiac death: a case report. AB - Simultaneous pancreas and kidney transplantation (SPK) is the treatment of choice for insulin-dependent diabetics with end-stage renal failure. The shortage of suitable pancreata has led to the use of donors after cardiac death (DCD). Although more than 20,000 pancreas transplants have been performed worldwide, the experience of pancreas transplantation from DCD remains limited. Factors during recovery, storage, and reperfusion may result in injury to the duodenal segment of the pancreas allograft. Here we have reported a case of duodenal necrosis after reperfusion in a patient who underwent SPK from a DCD. The pancreas was salvaged by excising the duodenum and performing a direct pancreas duct to bladder anastomosis. After recovery of function by the transplanted kidney, the patient developed recurrent urinary sepsis. The bladder-drained pancreas was converted to a direct enteric drainage, which resolved the recurrent urinary sepsis. PMID- 19545727 TI - Interleukin-1 receptor antagonist enhances islet engraftment without impacting serum levels of nitrite or osteopontin. AB - Interleukin-1beta (IL-1beta)-mediated early islet graft dysfunction and loss of islet mass can occur in different phylogenic types of islet transplantation. Large quantities of interleukin-1 receptor antagonist (IL-1RA) have been demonstrated to impede IL-1beta-mediated adverse effects on islet grafts in allo- and xenotransplantation. To clarify the influence of IL-1RA on early function and mass change, as well as long-term hypoglycemic effects of islet isografts, we studied streptozotocin-induced diabetic C57BL/6 mice infected with replication defective adenovirus carrying the mouse IL-1RA cDNA gene. This vector increased the mean serum level of IL-1RA to 8 ng/mL, approximately three times greater than for mice receiving adenovirus carrying the beta-galactosidase (beta-Gal) gene. The blood glucose levels declined faster and the insulin content of the graft was significantly higher on day 10 following transplantation among mice receiving mIL 1RA gene than the controls. Nevertheless, the insulin content of the pancreatic remnant did not differ among mice in the IL-1RA, beta-Gal, and vehicle control groups. Serum levels of nitrite and osteopontin before and 3 days after islet transplantation did not differ considerably among the IL-1RA, beta-Gal, and vehicle groups. Compared with the beta-Gal group, temporary posttransplantation hyperglycemia was significantly shortened in the IL-1RA group mice. Removal of graft-bearing kidneys at 13 weeks following transplantation caused recurrence of hyperglycemia in all treated diabetic mice. The insulin content of pancreatic remnants removed at 15 weeks following transplantation was similar in the IL-1RA and beta-Gal groups. In conclusion, a mildly elevated serum concentration of IL 1RA protected and enhanced engraftment of islet isografts immediately after transplantation. PMID- 19545728 TI - AMT, an inducible nitric oxide synthase inhibitor, enhances islet engraftment. AB - We co-transplanted silica gel-entrapping 4H-1,3-Thiazin-2-amine,5,6-dihydro-6 methyl monohydrochloride (AMT) with islets to evaluate the effects of AMT on early graft dysfunction in a syngeneic mouse model. The mean diameter of AMT embedding silica gel particles was 595 +/- 275 nm. The cumulative release of AMT was 29% at 1 hour and 45% at 72 hours. Sixteen streptozotocin-induced diabetic mice were separated into 3 groups. Group A received 50 islets (n = 4). Group B received 50 islets and blank silica gel (n = 6). Group C received 50 islets plus silica-gel containing 6.4 microg AMT (n = 6). Mice in group C required significantly less time for temporary posttransplantation hyperglycemia than those in groups A and B (A, 39 +/- 7 vs B, 40 +/- 5 vs C, 24 +/- 2 days; P < .05). The insulin contents of grafts retrieved at 13 weeks were 1.17 +/- 0.11 (n = 4), 1.01 +/- 0.16 (n = 6), and 1.68 +/- 0.30 microg (n = 6) for mice in groups A, B, and C, respectively. Pancreatic remnant insulin did not differ significantly between the 3 groups (A, 0.32 +/- 0.04 [n = 4] vs B, 0.29 +/- 0.06 [n = 6] vs C, 0.40 +/- 0.05 microg [n = 6]; P > .05). In vitro study revealed that 4 and 20 nmol/L of sol-gel-embedded AMT protected 87% and 96% RIN-m5F cells from 1 ng/mL interleukin-1beta-mediated destruction, respectively. Silica-gel entrapped AMT protects islet graft from a nonspecific inflammatory destruction, which is partly mediated via interleukin-1beta. PMID- 19545730 TI - Effects of insulin-like growth factor-1 and donor age on transplantation of porcine neonatal pancreatic cell clusters. AB - Porcine neonatal pancreatic cell clusters (NPCCs) isolated from 1- to 3-day-old pigs cured diabetic nude mice more than 14 weeks after transplantation. To shorten the latent period between transplantation and reversal of hyperglycemia, we investigated the effects of insulin-like growth factor-1 (IGF-1) and NPCCs isolated from 1-month-old pigs after transplantation. Pig pancreata were cut into fragments, collagenase digested, and then cultured. Three hundred and 2000 NPCCs were transplanted under the kidney capsule of nondiabetic and diabetic nude mice, respectively. After transplantation, the graft-bearing kidneys were removed to measure insulin content. NPCCs isolated from 1- to 3-day-old pigs were cultured with or without IGF-1 for 6 days. The stimulation index was not significantly different between the 2 groups at 1, 2, or 4 weeks. Moreover, at 4 weeks after transplantation of 300 NPCCs to nondiabetic nude mice yielded comparable graft insulin content as the recipients of NPCCs precultured with or without IGF-1. Two thousand cultured NPCCs isolated from 1-to 3-day-old pigs or 1-month-old pigs were transplanted into diabetic nude mice. The blood glucose levels of diabetic recipients in both groups decreased at the same rate after transplantation, achieving normoglycemia at 8 weeks. The graft insulin content at 12 weeks was not different between the 2 groups. Our data indicated that isolated NPCCs cultured with IGF-1 showed no beneficial effects on insulin secretion and transplantation; NPCCs isolated from 1-to 3-day-old and 1-month-old pigs displayed similar effects on transplantation. PMID- 19545729 TI - Polyglycolic Acid-islet grafts improve blood glucose and insulin concentrations in rats with induced diabetes. AB - Pancreatic islet transplantation is a promising therapeutic treatment for type 1 diabetes mellitus. In the present study, we cocultured islets with or without a polyglycolic acid (PGA) fibrous scaffold for 5 days and transplanted the PGA islet grafts into the leg muscles of Wistar rats with streptozotocin-induced diabetes; controls were injected with saline. The results showed that the blood glucose concentrations of the group given islets embedded with the PGA scaffold were lower than those without the scaffold or controls. On the other hand, the insulin content of the PGA-islet group was higher at all 5 time points compared with the insulin contents of the other 2 groups. After transplantation, many islets in the PGA-islet grafts showed normal morphology (as seen under the scanning electron microscope) and were surrounded by red blood cells. A fibrous extracellular matrix was visible around the PGA-islet grafts. These results demonstrated that PGA-islet grafts improved blood glucose and insulin concentrations in rats with induced diabetes. PMID- 19545731 TI - Bone marrow-derived mesenchymal stem cells improve islet graft function in diabetic rats. AB - Type 1 diabetes is associated with a progressive loss of beta cells and pancreatic islet transplantation could represent a cure for this disease. Herein we explored whether transplantation of bone marrow-derived mesenchymal stem cells (MSCs) allowed a reduced number of pancreatic islets to improve glycemic control in diabetic rats, by promoting islet vascularization. We transplanted 2000 syngenic islets alone or in combination with MSCs (10(6) cells) under the kidney capsules of diabetic Lewis rats. Animals transplanted with 2000 islets never reached normoglycemia. In contrast, rats transplanted with 2000 islets plus MSCs, showed a gradual fall in glycemia after transplantation, with normoglycemia maintained until killing. Comparable glycemic control was obtained with transplantation of 3000 islets alone. The MSC preparation used for in vivo experiments expressed high levels of vascular endothelial growth factor (VEGF(165)) and, at less extent, VEGF(189), as evaluated by reverse transcriptase polymerase chain reaction (RT-PCR). In transplanted animals, vascularization was quantified by morphometric analysis of islet grafts with anti-RECA and anti insulin antibodies. MSCs were stained with PKH-26. Mean capillary density was 1002 +/- 55 capillaries/mm(2) in islets transplanted alone. Co-infusion of MSCs with islets significantly increased the number of capillaries to 1459 +/- 66 capillaries/mm(2). In conclusion, our study indicated that co-transplantation of MSCs with pancreatic islets improved islet graft function by promoting graft vascularization. PMID- 19545733 TI - Effects of pancreas cold ischemia on islet function and quality. AB - We used a rat model of pancreas cold preservation to assess its effects on islets. Glands were surgically retrieved and stored in University of Wisconsin (UW) solution for 3 hours (Short) or 18 hours (Long) cold ischemia time (CIT). Islet yield was significantly lower in the Long-CIT than the Short-CIT group, as well as islet recovery after overnight culture (P < .01). Islet cell viability after isolation was significantly reduced in the Long-CIT group (P < .05). Reversal of diabetes following transplantation of suboptimal islet grafts occurred earlier in the Short-CIT group than the Long-CIT. All animals in the Short-CIT group and 80% in the Long-CIT group achieved euglycemia. Freshly isolated islets showed a significant increase of JNK and p38 (P < .05) phosphorylation in Long-CIT compared with Short-CIT. Histopathological assessment of the pancreas showed a significantly higher injury score. Proteomic analysis of pancreatic tissue led to identification of 5 proteins consistently differentially expressed between Short-CIT and Long-CIT. Better understanding of the molecular pathways involved in this phenomenon will be of assistance in defining targeted interventions to improve organ use in the clinical arena. PMID- 19545732 TI - Time point is important for effects of syngeneic bone marrow transplantation for type 1 diabetes in mice. AB - OBJECTIVES: Autologous hematopoietic stem cell transplantation (HSCT) has recently become a novel therapy for patients with new-onset type 1 diabetes (T1D). However, the optimal time points for HSCT are still unknown. METHODS: By using multiple low-dose streptozotocin (STZ)-induced T1D mice models, we performed syngeneic bone marrow transplantation (syn-BMT) in diabetic mice at various time points new-onset day 3 (n = 12); new-onset day 10 (n = 13); later onset day 20 (n = 12); and day 40 (n = 7), respectively. At 120 days after syn BMT, we examined pancreata histology, serum insulin, and CD4(+)CD25(+)FoxP3(+) T regulatory lymphocytes (Tregs). RESULTS: Our previous results showed that syn-BMT can overcome diabetes when performed on day 10, but not at day 40. Our new data showed BMT only attenuated diabetes when done on day 3 or day 20. Moreover, the percentage of Tregs in the spleen correlated with the attenuation of hyperglycemia. SIGNIFICANCE: These results indicated that syn-BMT should be performed, when diabetes is neither too new-onset nor too late-stage. Tregs represent one mechanism for syn-BMT-induced restoration of immune tolerance in STZ-diabetic mice. PMID- 19545734 TI - Panel-reactive screening and treatment practices following bridge to transplantation ventricular assist device placement. AB - The use of left ventricular assist devices (LVAD) as a bridge to transplantation is associated with the development of circulating antibodies. We conducted a survey of all adult cardiac transplantation programs in the United States in an attempt to define current practices with regard to LVAD implantation, monitoring panel-reactive antibody (PRA) levels, treatment options, and peritransplantation management. Pretransplantation sensitization with the use of LVAD is a concern to the majority of transplantation professionals and there is no consensus on the need or mode of treatment. PMID- 19545735 TI - Efficacy of total lymphoid irradiation in azithromycin nonresponsive chronic allograft rejection after lung transplantation. AB - Bronchiolitis obliterans syndrome (BOS) remains a major problem after lung transplantation. Azithromycin seems to be beneficial in some patients with established BOS. We investigated the efficacy of total lymphoid irradiation (TLI) in 6 BOS patients with a continuous decline in FEV(1), despite treatment with azithromycin for a mean of 12 +/- 13 (range, 1-35) months. A historical control group consisted of 5 patients with declining FEV(1), also nonresponders to azithromycin and those not treated with TLI. All 6 TLI patients received the total dose of 8 Gy in 10 sessions. There was a significant change in the decline of the FEV(1) after TLI treatment (from 221 +/- 107 to 94 +/- 79 mL/mo; P = .041). Three patients died, due to BOS progression, overwhelming pneumonia, and sudden cardiac arrest, respectively, 3.5, 11, and 26 months after TLI; two patients underwent retransplantation at 6 and 19 months after TLI, respectively. The sixth patient remains stable in BOS stage 3 after a follow-up period of 24 months. In the control group, there was no significant change in FEV(1) decline (209 +/- 97 mL/mo before versus 193 +/- 81 mL/mo after starting azithromycin; P = not significant). Two patients remain stable in BOS stage 3, 1 died of BOS progression, and the 5th patient is scheduled for retransplantation. We conclude that patients who do not or no longer respond to azithromycin may benefit from TLI, as suggested by a decreased rate in decline of the FEV(1). PMID- 19545736 TI - Tacrolimus versus cyclosporine for adult lung transplant recipients: a meta analysis. AB - BACKGROUND: Tacrolimus and cyclosporine are the 2 major immunosuppressants for lung transplantation. Several studies have compared these 2 drugs, but the outcomes were not consistent. The aim of this meta-analysis of randomized controlled trials (RCTs) was to compare the beneficial and harmful effects of tacrolimus and cyclosporine as the primary immunosuppressant for lung transplant recipients. METHODS: We conducted searches of electronic databases and manual bibliographies. We performed a meta-analysis of all RCTs comparing tacrolimus with cyclosporine as primary immunosuppression for lung transplant recipients. Extracted, pooled data for mortality, acute rejection, withdrawals, and adverse events were analyzed using Mantel-Haenszel tests with a random effects model. RESULTS: Three RCTs including 297 patients were assessed in this study. Mortality at 1 year or more was comparable between lung recipients treated with tacrolimus and cyclosporine (odds ratio [OR], 0.94; 95% confidence interval [CI], 0.42-2.10; P = .88). Tacrolimus-treated patients experienced fewer incidences of acute rejection (MD = -0.14; 95% CI, -0.28 to -0.01; P = .04). Pooled analysis showed a trend toward a lower risk of bronchiolitis obliterans syndrome (BOS) among tacrolimus-treated patients, although it did not reach significances (OR, 0.53; 95% CI, 0.25-1.12; P = .10). Fewer patients stopped tacrolimus than cyclosporine (OR, 0.12; 95% CI, 0.03-0.48; P = .003). The rate of new-onset diabetes was higher among the tacrolimus group (OR, 3.69; 95% CI, 1.17-11.62; P = .03). The incidence of hypertension and renal dysfunction were comparable in these 2 groups (OR, 0.24; 95% CI, 0.03-1.70; P = .15; and OR, 1.67; 95% CI, 0.70-3.96; P = .25, respectively). There was a trend toward lower risk of malignancy in tacrolimus treated patients, although it did not reach significance either (OR, 0.19; 95% CI, 0.03-1.13; P = .07). The incidence of infection was comparable in these 2 groups (MD = -0.29, 95% CI, -0.68 to 0.11; P = .16). CONCLUSION: Using tacrolimus as primary immunosuppressant for lung transplant recipient resulted in comparable survival and reduction in acute rejection episodes when compared with cyclosporine. PMID- 19545737 TI - Resolution of refractory chronic autoimmune thrombocytopenic purpura following mesenchymal stem cell transplantation: a case report. AB - Herein we have reported the case of a man with chronic autoimmune thrombocytopenia (AITP) refractory to conventional therapy who underwent T-cell depleted autologous peripheral blood stem cell (PBSC) transplantation. A complete, steroid-independent, platelet remission was obtained, but unfortunately he subsequently relapsed after 3 months. Finally, we administered human adipose tissue-derived mesenchymal stem cells (AMSC) with resolution to the AITP. PMID- 19545738 TI - Interindividual variation of maximal blood levels of tacrolimus after its oral administration in hematopoietic cell transplant recipients. AB - We investigated the pharmacokinetics of oral tacrolimus in 31 hematopoietic cell transplant recipients, identifying 2 subgroups based on the differences between C(0) (trough) and C(max) (maximal) levels: group A (n = 21; 68%) with a C(max) C(0) value of <10 ng/mL, and group B (n = 10; 32%) with a C(max)-C(0) value of >or=10 ng/mL. Although the C(0) and C(12) values were not significantly different between the 2 groups, the mean area under the concentration curve for 12 hours (AUC(0-12)) was significantly greater in group B than group A (200.9 +/- 36.3 vs 155.1 +/- 43.1 ng.h/mL; P < .05), and the mean half-life was significantly shorter in group B than group A (13.55 +/- 6.70 vs 18.17 +/- 6.30 hours; P < .05). Thus after the oral administration of tacrolimus, we observed a notably high AUC due to high peak level, which we were unable to predict simply by measuring the trough level. A pharmacokinetic analysis of each patient was essential to optimize the oral tacrolimus dose. PMID- 19545739 TI - Immunosuppressive effects of emodin: an in vivo and in vitro study. AB - OBJECTIVE: We sought to investigate the immunosuppressive effects of emodin and its potential in vivo and in vitro mechanisms. METHODS: In vitro immunosuppressive effects of emodin were analyzed by its ability to suppress the response of human peripheral blood mononuclear cells to phytohemagglutinin (PHA) and to mixed lymphocyte culture (MLC). We examined changes in interleukin (IL)-2 and IL-4 in within MLC supernates. The in vivo immunosuppressive effects of emodin were analyzed using a skin transplantation model in mice. We also investigated the mean survival time (MST) and plasma IL-2 levels. RESULTS: In vitro experiments: Responses of mononuclear cells to PHA and MLC were suppressed by emodin treatment. Decreased production of IL-2 along with promoted secretion of IL-4 was also observed by emodin treatment during MLC. In vivo experiments: The emodin-treated group showed prolonged MST of skin grafts and decreased serum IL-2 production. CONCLUSIONS: Emodin showed immunosuppressive activities both in vivo and in vitro. The potential immunosuppressive mechanism of emodin's may be suppression of lymphocyte proliferation and influences on cytokines. PMID- 19545740 TI - Transferable cardiac allograft acceptance induced by transfusion of donor B cells with impaired inducible costimulator/B7h allorecognition. AB - OBJECTIVE: Donor-specific transfusion (DST) of leukocytes with an impaired costimulatory signal has been proven to be an effective way to improve allograft survival. Inducible costimulator (ICOS) has been shown to play a crucial role in acute and chronic allograft rejection. To test the role of ICOS signaling during DST, we employed ICOS-Fc-targeted B cells as antigen of DST to challenge the allogeneic engraftment in vivo. MATERIALS AND METHODS: A murine cardiac allograft model was employed using BALB/c donors and C57BL/6 recipients, while various transfusions were performed according to treatment protocols. RESULTS: Allograft survival was prolonged by infusion of ICOS-Fc-targeted B cells; however, allograft acceptance could not be achieved unless additional systemic injections of ICOS-Fc were given. Adoptive transfer of splenic CD4(+) but not CD4(+)CD25(-) subsets from long-term allograft survival (LTAS) mice to lightly irradiated naive recipients resulted in subsequent BALB/c allograft acceptance without additional immunosuppression. CONCLUSIONS: ICOS/B7h signaling during direct allorecognition played an important role in prolonging allograft survival, and an allograft acceptance can be established by DST with complete blockade of ICOS/B7h in both direct and indirect allorecognition. Interestingly, this allograft acceptance was transferable and maintained at least partly by the immune regulation of CD4(+)CD25(+) T cells. These findings may help to design a potential therapeutic treatment to prevent allograft rejection by DST in combination with ICOS/B7h blockade. PMID- 19545741 TI - Tolerant T cells inhibit natural killer cells function in antigen-presenting cells in an independent fashion. AB - Understanding the relationships among immune cells in the setting of immunologic tolerance is imperative to maintain organ and tissue transplants. T cells and natural killer (NK) cells are responsible for both immune tolerance and immune rejection; however, there is only limited knowledge about the relevance of T and NK cells in tolerance. To address this issue, we explored the possible actions of tolerant T cells on NK cells by the means of mixed lymphocyte co-cultures and NK cytotoxicity assays. We showed that tolerant T cell-induced blockade of the co stimulatory pathway significantly inhibited NK cell function in vitro regarding antigen-presenting cells. This action was cell-cell-contact dependent. We argue that tolerant T cells and NK cells impart synergistic cooperation to maintain transplant tolerance. PMID- 19545742 TI - Use of epigenetic modification to induce FOXP3 expression in naive T cells. AB - We investigated whether epigenetic modification agents can convert naive T cells to regulatory T cells (T(regs)) which are responsible for limiting immune responses and maintaining self-tolerance. We treated splenic CD4(+)/CD25(-) naive T cells from BALB/c mice with the DNA-methyltransferase inhibitor 5-aza-2' deoxycytidine (5AzaD) or the histone protein deacetylase (HDAC) inhibitor Trichostatin A (TSA), and measured the effects on the expression of FOXP3, which encodes a transcription factor (FOXP3) that regulates T(reg) development. FOXP3 expression in naive T cells was increased by 5AzaD or TSA treatment, administered 72 hours after T-cell receptor (TCR) stimulation with anti-CD3 plus anti-CD28. The T(regs) induced by 5AzaD or TSA expressed greater amounts of the FOXP3 protein than the control and the natural T(regs). The analysis of T(reg) associated markers also showed T(reg) phenotypes (CD25(+)/CTLA4(+)/GITR(+)/CD127( )). Finally, the induced T(reg) population also displayed T-cell suppression. These data suggested that epigenetic modification agents can induce FOXP3 expression, promoting the conversion of naive T cells to T(regs). PMID- 19545743 TI - Arsenic trioxide attenuated the rejection of major histocompatibility complex fully-mismatched cardiac allografts in mice. AB - We investigated the effects of arsenic trioxide (As(2)O(3)) on allogeneic immune response using a mouse heart transplantation model. Mice were randomly divided into 4 groups of 6 animals each. The control group received phosphate-buffered saline (PBS); the As(2)O(3)-treated group, intraperitoneal (IP) injection of As(2)O(3) (1 mg/kg) from days -3 to 10 after heart transplantation. The cyclosporine (CsA)-treated group was given a subtherapeutic dose of CsA (10 mg/kg) IP, and the As(2)O(3) plus CsA-treated group, a combined protocol of As(2)O(3) and CsA. Six days after transplantation, cardiac allografts were harvested for immunohistology and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. The survival of the allografts was significantly improved among the As(2)O(3)-treated group compared with the control group (17.2 +/- 1.9 vs 8.0 +/- 0.9 days; P < .05). A marked prolongation (28.6 +/- 6.0 days) of graft survival was achieved by the combined protocol compared with the CsA-treated group (9.6 +/- 3.0 days; P < .05) or the As(2)O(3)-treated group. Allografts of As(2)O(3)-treated and As(2)O(3) plus CsA-treated mice showed a changing pattern of Th1/Th2 cytokine mRNA expression. Allograft rejection was apparently alleviated by low-dose As(2)O(3), and particularly when combined with a subtherapeutic CsA dose. PMID- 19545744 TI - The effect of Malononitrilamides (FK778) on phenotypic properties of human peripheral dendritic cells. AB - BACKGROUND: FK778, a malononitrilamide analogue of lefunomide, is currently a promising immunosuppressive drug. Because the cellular and molecular mechanisms underlying the effects of FK778 are not entirely clarified. We studied its effects on human peripheral dendritic cells. METHODS: Peripheral blood mononuclear cells (PBMC) from 12 healthy volunteers were isolated by density separation over Ficoll solution. After resuspension in adaptive immunotherapy medium (AIM)-V medium, they were cultured without exogenous growth factors. The study group was treated with FK 778 (50 microg/mL) or Rapamycin (10 ng/mL). The phenotype of dendritic cell was ascertained by indirect immunoflurescence for analysis by flow cytometry. RESULTS: Compared with the Rapamycin-treated controls, the expressions of CD80, CD83, CD86, HLA-DA, CD54, CD62, CCR5, and CCR7 in the FK778-treated myeloid dendritic cells and the expression of CD80, CD83, CD86, HLA-DA, and CD54 in the FK778-treated plasmacytoid dendritic cells were significantly down-regulated. CONCLUSION: FK778 inhibited the differentiation and maturation of dendritic cells. PMID- 19545745 TI - Adenovirus-mediated CTLA4Ig gene transfer improves the survival of grafted human hepatic progenitors in mouse liver. AB - The invasive nature of surgery and limited numbers of donor livers for end-stage patients has prompted the search for alternative cell therapies for intractable hepatic disease. Hepatocyte transplantations have been performed for a variety of indications, but sustained benefits have not been observed in most cases. Rat fetal liver epithelial cells (liver stem cells) have demonstrated self-renewal in vivo and functional repopulation of the liver. We have previously isolated and expanded epithelial progenitor cells (EPC) from the human fetal liver to investigate their differentiation potential. In this study, we applied suppression of immunorejection by adenoviral CTLA4Ig gene delivery mediated to examine the survival and differentiation of human fetal EPC transplanted into normal mouse liver. The grafted EPC showed extensive proliferation at both 1 and 2 months after transplantation compared with controls. Moreover, most EPC differentiated into hepatocytes, while a small fraction became bile ductular cells. This finding suggested that human fetal EPC may be a ideal source of cell based therapy for various liver diseases. PMID- 19545747 TI - Effects of long-term FK 506 therapy on the alveolar bone and cementum of rats. AB - Cyclosporine (CsA) and tacrolimus (FK 506) exert complex, incompletely understood actions on bone. The objective of the study was to evaluate the effects of long term tacrolimus therapy on the periodontium. Rats were treated for 60, 120, 180, and 240 days with daily subcutaneous injections of 1 mg/kg body weight of FK 506. After the experimental period, we obtained serum levels of calcium and alkaline phosphatase (ALP). After histological processing, the alveolar bone and cementum, as well as volume densities of bone (V(b)) and osteoclasts (V(o)), were assessed at the regions of the lower first molar. There was a tendency toward a statistically significant decrease in ALP levels with FK 506; however, serum calcium levels increased during the long periods. At 60, 180, and 240 days of treatment with FK 506, we did not observe V(b) and V(o) alterations. At 120 days of treatment, there was an evident decrease in V(b), but it did not show alveolar bone loss. We did not observe any alterations of cementum among rats treated with FK 506. It may be concluded that FK 506 administration did not induce side effects on the periodontium. PMID- 19545746 TI - Effect of cyclosporine and sirolimus on fatty acid desaturase activities in cultured HEPG2 cells. AB - The aim of the present work was to evaluate the influence of cyclosporine (CsA) and sirolimus (SRL) on fatty acid (FA) desaturase activities. These enzymes (named Delta9, Delta6, and Delta5 desaturases) catalyze reactions leading to the biosynthesis of n-9, n-6, and n-3 FA families. n-3 FA family, derived from alpha linolenic acid, is involved in the prevention of vascular events, which appear after successful kidney transplantation. Five groups of HepG(2) cells in culture were treated with either CsA (1 microg/microL and 2 microg/microL) or SRL (10 ng/mL and 20 ng/mL) for 3 days, including a control group without immunosuppressive treatment. We studied the incorporation and metabolic conversion of radioactive [1-(14)C]palmitic, linoleic, and eicosatrienoic acids. We also analyzed fatty acid composition. The distribution of radioactive metabolic products after incubation of these cells with [1-(14)C]palmitic acid revealed a decrease in Delta9 desaturase activity in the presence of each immunosuppressive drug: CsA = 0.61 +/- 0.01; SRL = 0.59 +/- 0.04 versus control = 0.79 +/- 0.05 (P < .01). We observed a significant increase in Delta6 and Delta5 desaturase activities under the influence of the immunosuppressive drugs: radiolabeled linoleic acid (CsA: 0.93 +/- 0.04; SRL: 1.02 +/- 0.03 vs control 0.60 +/- 0.03; P < .01) and eicosatrienoic acid (CsA: 1.12 +/- 0.02; SRL: 1.07 +/ 0.01 vs control 0.75 +/- 0.01; P < .01). In conclusion, CsA and SRL modulated the biosynthesis of polyunsaturated FAs, decreasing Delta9 desaturase and increasing Delta6 and Delta5 desaturase activities. PMID- 19545748 TI - Melatonin ameliorates tacrolimus (FK-506)'s induced immunosupressive effect in rat liver. AB - Tacrolimus (FK-506) is a powerful immunosuppressive agent that modulates neutrophil infiltration during inflammation. In this study, we sought to investigate the effects of melatonin on malondialdehyde (MDA), nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor-alpha (TNF-alpha) to oppose the negative effects of the immunosupressant FK-506. Group A was sham; group B, tacrolimus (1 mg/kg/d subcutaneously); and Group C received tacrolimus (1 mg/kg/d plus melatonin). All tissues underwent histopathologic examination. The MDA level in group B increased 53% compared with the sham group (P < .001); in group C, the MDA level decreased 16% compared with group B (P > .05). While TNF-alpha in group B increased 68.8% compared with the sham group (P < .001) and in group C it decreased 63.5% compared with the sham group. The IL-6 level in group B increased 81%; in group C, it decreased 13% compared with group B. NO levels in group B increased 48% compared with the sham group and increased by 15% in group B compared with group C. Thus melatonin may serve as a protective agent against the side effects of tacrolimus. PMID- 19545749 TI - Mesenchymal stem cells treated with rat pancreatic extract secrete cytokines that improve the glycometabolism of diabetic rats. AB - BACKGROUND: Mesenchymal stem cells (MSCs) under favorable conditions secrete a spectrum of cytokines that promote the survival of surrounding cells via paracrine mechanisms. We explored the impact of rat pancreatic extract (RPE) on cytokine secretion by MSCs and examined the influence of administration of conditioned media of MSCs treated with RPE on blood glucose levels in diabetic rats. METHODS: Cytokine levels (IGF-1, VEGF, bFGF) in conditioned media of MSCs treated with RPE were measured using enzyme-linked immunosorbent assays. We estimated blood glucose levels of STZ-induced diabetic rats following intraperitoneal injection of conditioned media from RPE-treated MSCs. We analyzed histopathology of pancreatic islets by insulin immunostaining and apoptosis through a TUNEL assay. RESULTS: Levels of IGF-1, VEGF, and bFGF were significantly increased in RPE-CM compared with control media. Administration of conditioned media of RPE-treated MSCs significantly lowered the blood glucose levels of diabetic rats. After RPE treatment the insulin-positive area was increased and apoptosis of pancreatic beta cells decreased. CONCLUSION: RPE enhanced the secretion of cytokines by MSCs. MSCs in the pancreatic microenvironment may exert indirect salutary effects via paracrine mediators on injured pancreatic cells in an STZ-induced diabetic animal model. The secreted factors may exert their therapeutic benefits by preventing apoptosis of pancreatic beta cells. PMID- 19545750 TI - Development of functional human immune system with the transplantations of human fetal liver/thymus tissues and expanded hematopoietic stem cells in RAG2-/ gamma(c)-/- MICE. AB - BACKGROUND: There is an increasing need for suitable animal models for the study of the human immune system and disease. The purpose of this study was to develop a practical in vivo model of human immune cell repopulation using ex vivo expanded human fetal liver-derived CD34(+) hematopoietic stem cells and subrenally coimplanted fetal liver/thymus tissues. METHODS: Freshly isolated fetal liver-derived CD34(+) hematopoietic stem cells were frozen until injected and ex vivo expanded with various cytokines for 7 days. After fetal liver/thymus tissues were subrenally coimplanted into preirradiated Rag2(-/-)gamma(c)(-/-) mice, frozen and ex vivo expanded CD34(+) cells were injected intravenously. The peripheral blood of the mice was monitored for the detection of human cell engraftment using flow cytometry. Then we confirmed human T-cell function by in vitro function assays. RESULTS: After fetal liver/thymus tissues were coimplanted into the irradiated Rag2(-/-)gamma(c)(-/-) mice, with frozen and ex vivo expanded CD34(+) hematopoietic stem cells, human cell engraftments were determined using hCD45 and multilineage markers. The cultured cells with the cytokine combination of stem cell factor, thrombopoietin, Flk2/Flk3 ligand (FL), and interleukin-3 showed stable and long-term engraftment compared to other combinations. The ex vivo expanded human fetal liver-derived CD34(+) hematopoietic stem cells, under our culture conditions, accomplished a large volume of expanded cells that were sustained, demonstrating self-renewal of the evaluated markers, which may have indicated long- term repopulation activity. CONCLUSION: The results of this study demonstrated a practical mouse model of expanded human immune cells especially T cells in Rag2(-/-)gamma(c)(-/-) mice. PMID- 19545751 TI - Evaluation of stem cell differentiation in diabetic rats transplanted with bone marrow mesenchymal stem cells. AB - OBJECTIVE: Bone marrow mesenchymal stem cell (BM-MSC) transplantation has generated a great deal of excitement as a promising therapeutic strategy for diabetes mellitus. However, the exact mechanisms of reversing hyperglycemia remain elusive. Our objective was to investigate whether stem cell differentiation determined therapeutic efficacy. MATERIALS AND METHODS: Wistar rats were rendered diabetic by an intraperitoneal injection of streptozotocin. BM MSCs isolated from diabetic Wistar rats were analyzed for phenotype characteristics. Subsequently, BM-MSCs were transplanted into diabetic rats, followed by intravenous injection of recombinant lentiviruses encoding 2 different small hairpin RNAs (shRNAs) for specific interference with neurogenin 3 (Ngn3). We measured blood glucose levels and insulin and performed histological analysis of the pancreas. RESULTS: BM-MSCs lowered blood glucose by increasing beta-cell mass compared with sham-operated controls, but this effect was inhibited by interference with the Ngn3 gene. CONCLUSION: Differentiation of stem cells, including BM-MSCs and endogenous pancreatic stem cells, plays a major role in the process of reversing hyperglycemia. PMID- 19545752 TI - Induction of mixed chimerism in mice by employing different conditioning protocols and bone marrow cell transplantation. AB - Mixed chimerism has been suggested to produce allograft tolerance. Since this phenomenon is not fully understood, the aim of our study was to evaluate various protocols for chimerism induction in a mouse model. B6.SJL-Ptprc(a)Pep3(b) mice were injected with 20 to 30 x 10(6) bone marrow cells from Balb C mice. Conditioning consisted of total body gamma irradiation with 9.5, 5, and 3 Gy on " 1 day" of the experiment, with 200 mg/kg cyclophosphamide (CP) ("+2 day"). Additionally, one group of mice received blocking antibody against CD40L on days 0, 1, 4, and 7. The presence of mixed chimerism in peripheral blood was assessed at 1, 2, 3, 4, 6, and 8 weeks using flow cytometry to detect CD45.1 or CD45.2 antigen expression. Moreover, the chimerism was examined in CD4, CD8, CD45/B220, Mac-1alpha subpopulations in peripheral blood and bone marrow cells at 8 weeks. We also compared chimerism in peripheral blood, bone marrow, and spleen leukocyte populations. We observed that the most effective conditioning method with relatively low toxicity was based on concomitant use of 5 Gy total body irradiation and CP. The percentage of donor cells differed among peripheral blood subpopulations and bone marrow cells, but was similar in leukocyte populations derived from various sources. Our experiments sought to optimize the induction of stable mixed chimerism. PMID- 19545753 TI - Gene transfer of calcitonin gene-related peptide suppresses development of allograft vasculopathy. AB - OBJECTIVE: To explore suppression of allograft vasculopathy by transfer of the calcitonin gene-related peptide (CGRP). METHODS: The descending thoracic aortas from Lewis rats were grafted to the abdominal aortas of F344 rats, and the rats were randomized into 2 groups. A gene construct containing sequences from the adenoviral oncoprotein, the CGRP, and the enhanced green fluorescent protein was transferred into 1 group, and the sequences for the adenoviral oncoprotein and enhanced green fluorescent protein were transferred into a control group. Specimens were harvested at 4 and 8 weeks. Gene transfer was confirmed at fluorescence microscopy of frozen tissue sections, and expression was measured using reverse transcriptase-polymerase chain reaction. We determined the locations and levels of vascular cell adhesion molecule-1 (VCAM-1) and inducible nitric oxide synthase (iNOS) at immunohistochemistry and measured apoptosis. RESULTS: The CGRP gene was expressed only in the CGRP group at 4 weeks. The vascular luminal occlusion score in the CGRP group was lower than in the control group. The apoptotic index of the CGRP group was lower than in the control group only at 4 weeks. The VCAM-1 immunohistochemistry score in the CGRP group was lower than in the control group; however, the iNOS immunohistochemistry score in the CGRP group was lower than in the control group in the intima only at 4 weeks. CONCLUSION: The expression of CGRP effectively suppressed the development of allograft vasculopathy and encroachment by lymphocytes and inflammatory cells. This reduced the levels of VCAM-1 to inhibit apoptosis induced by iNOS; thus, the tissue of the allografted vessel was protected and rejection was averted. PMID- 19545754 TI - Biomechanical study of hepatic portal vein in humans and pigs and its value in liver transplantation. AB - OBJECTIVE: Our aim was to explore the biomechanical properties of hepatic portal vein (HPV) in humans and pigs to provide evidence for liver xenotransplantation. MATERIALS AND METHODS: The pressure-diameter relationships of HPV from 6 deceased donors and 36 pigs were measured on a biomechanical experimental stand to calculate the elastic modulus and compliance. Each sample sliced into 5-mm frozen sections was stained with hematoxylin-eosin (H&E). Geometric morphological indices were measured with a computer image analysis system. RESULTS: The length, wall thickness, and diameters of HPV in pigs increased from 1 to 6 months (P < .05). There were no significant differences between 6-month-old pigs and adult humans (P > .05). The incremental elastic modulus of the pig HPV increased with age, whereas the compliance decreased. There was no difference in the elastic modulus of HPV between 5- to 6-month-old pigs and humans (P > .05). Also, there was no difference in HPV compliance between 6-month-old pigs and humans (P > .05). CONCLUSIONS: Our results suggested that the biomechanical properties of HPV in 6-month-old pigs were similar to those of humans. From a biomechanical perspective, anastomosis of corresponding HPV from 6-month-old pigs to humans may be feasible in the process of pig-to-human liver xenotransplantation. PMID- 19545755 TI - Cryopreserved porcine tendons preserve cell viability after thawing. AB - Controlled cryopreservation is an important method for storage of tissue grafts in skin banking, reproductive medicine and other domains. Although the availability of cryopreserved flexor tendons would be highly beneficial in reconstructive surgery, especially in complex reconstructions for which grafting material is limited, only a few studies have dealt with transplanted tendons. We achieved successful cryopreservation of porcine flexor tendons in 2 cryoprotective media: dimethyl sulfoxide and glycerol. Their viability was shown using a quantitative colorimetric MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl 2H-tetrazolium bromide) assay. For comparison of native and cryopreserved tendons (n = 7 samples each), the adopted viability index was the ratio of MTT-dependent optical density and tendon weight. The viability index of native samples did not change significantly after cryopreservation and thawing. The proliferative capacity of tendon fibroblasts after thawing was shown in primary cell cultures. The described cryopreservation protocol and MTT assay may provide a basis for future autografting of human tendons. PMID- 19545756 TI - Establishment of an acute graft-versus-host disease model following liver transplantation in donor-dominant one-way major histocompatibility complex matching rats. AB - Acute graft-versus-host disease is an uncommon but devastating complication following liver transplantation (LTx-aGVHD). We investigated whether a rat model of LTx-aGVHD could be established using Lewis rat donors and (LewisXBN)F1 rats as recipients, which provides favorable conditions for studies of graft-versus-host reaction or disease. Replacement of (LewisXBN)F1 livers by Lewis livers alone was not sufficient to induce aGVHD; all recipients grew in a normal pattern as the syngeneic liver transplantation (LT) from Lewis to Lewis rat. However, when various numbers of donor splenocytes (1 x 10(8), 2 x 10(8), 3 x 10(8), 4 x 10(8)) were transferred simultaneously with the LT, the morbidities of lethal aGVHD were 16.7%, 50%, 83.3%, and 100%, respectively. The clinical courses as well as histologic analyses of skin and colon showed typical aGVHD characteristics. However, unlike transfusion-associated aGVHD, the liver graft was not involved. These clinical and histologic characteristics of aGVHD were consistent with those in humans who develop aGVHD after LT. Thus, a reproducible rat model of LTx-aGVHD was developed by performing LT from Lewis to (LewisXBN)F1 rats in combination with donor splenocyte transfusion. This model may be useful for further studies on the mechanisms and effective treatment modalities for LTx-aGVHD. PMID- 19545757 TI - Investigation of blood typing method for seoul National University miniature pig. AB - Pig blood group antigens may be present on grafted tissue as 16 isoforms, including the major one, A substrance. Seoul National University (SNU) miniature pigs and domestic pigs were used in this study. Polymerase chain reaction (PCR) was performed for the erythrocyte antigen A (EAA) gene, and reverse transcriptase PCR for pig A transferase fucosyl transferase (FUT) 1 and FUT-2. The hemagglutination test was performed with murine monoclonal anti-A and anti-B antibodies (mAb), and immunohistochemistry with anti-human blood antigen mAb. SNU miniature and domestic pigs showed blood groups A and O. Blood group A SNU miniature pigs expressed either EAA(AA) or EAA(AO) and either S(SS) or S(SO); blood group O miniature pigs expressed EAA(OO) and S(SS) or S(SO), and there was no A(weak). Additionally, blood group A could be divided into blood group A(clotting) and blood group A(not clotting) in hemagglutination tests. Pig A substance was expressed in the lung and kidney in blood group A pigs, but we could not detect pig A substance expression in the lung, kidney, and heart of blood group O pigs or the heart of blood group A pigs. In conclusion, we suggest that blood typing of SNU miniature pigs can be easily performed using immunohistochemistry, PCR, and/or RT-PCR. Molecular-based AO typing described in this study may be useful to select SNU miniature pigs bearing a specific blood group. PMID- 19545758 TI - Allogeneic stimulation causes transcriptional reactivation of latent murine cytomegalovirus. AB - Cytomegalovirus (CMV) reactivation is a well-described complication of transplantation that may be caused by allogeneic stimulation, immunosuppression, or both. These studies were performed to determine if allogeneic stimulation alone is sufficient to reactivate latent CMV. BALB/c mice latently infected with Smith strain murine CMV (MCMV) received allograft (n = 8), allograft plus cortisol (n = 5), or isograft (n = 4) skin. All allograft recipients rejected their grafts within 9 to 12 days of transplantation. Three weeks after grafting, recipients were evaluated for MCMV reactivation, and all allograft recipients (8/8) showed MCMV reactivation, while no isografts had reactivation (0/4). Surprisingly, cortisol therapy blocked MCMV reactivation (0/5). These data suggested that allogeneic stimulation alone can trigger systemic reactivation of latent CMV. Although immunosuppression is thought to contribute to reactivation, certain agents that impair NF-kappaB activation may actually reduce reactivation. PMID- 19545759 TI - Primary cultures from rat vibrissae as a potential cell source for in vitro construction of urinary bladder wall grafts. AB - BACKGROUND: In vitro-constructed grafts can be used for human bladder augmentation. There are many diseases in which autologous cells cannot be used for this purpose. The aim of the present study was to examine the potential of rat vibrissae hair follicle cells to form cultures, which could serve as a source for in vitro creation of urinary bladder wall grafts. METHODS: Two hundred vibrissae were excised from young Wistar male rats. Two different digestions were performed, in dispase and in collagenase. All follicles were additionally incubated in trypsin and ethylenediamine tetraacetic acid. Two different culture media based on DMEM (Dulbecco's Modified Eagle's Medium) were used: the first was supplemented with keratinocyte growth factor (KGF) and the second with epidermal growth factor. Immunocytochemical detection of cytokeratin, CD34, p63, Ki-67 (proliferation index), and HMB45 (Human Melanoma Black 45) was performed. RESULTS: Forty-eight primary cultures of rat follicle vibrissae cells were established from 200 hair follicles (24% successful rate). Twenty-four primary cultures were obtained after dispase digestion and 24 after collagenase treatment. Each group was cultured in 2 different media. A heterogeneity of primary cultures was observed. Cells formed a monolayer within a period of 2 to 4 weeks. The 24 primary cultures established after dispase treatment exhibited monolayers of small cuboid cells expressing cytokeratin and CD34. In the 40th passage 20%-40% of cells expressed p63; 85% of these cells from late passages were positive for Ki-67, indicating preserved mitotic potential. Epithelial-like phenotype was observed after dispase digestion and cultivation in KGF supplemented medium. After 3 weeks, the morphology of these cells changed into fibroblast-like. These cultures were negative for CD34. Fibroblast-like cell growth was observed after collagenase treatment in both KGF- and EGF-supplemented media. These cells were positive for the melanocyte cell marker (HMB45). CONCLUSIONS: Culture media and isolation conditions influence hair follicle stem cell differentiation. The stem cell niche within the hair follicles is a reservoir of cells, which can be potentially used for in vitro creation of urinary bladder wall grafts. PMID- 19545760 TI - Urachal adenocarcinoma following kidney transplantation: the first case report. AB - A 53-year-old man who underwent successful kidney transplantation for stage 5 chronic kidney disease presented to our clinic with intermittent painless gross hematuria. Urachal adenocarcinoma, stage III A by Sheldon system, was diagnosed after serial histopathologic and radiological studies. The patient was treated with extended partial cystectomy, en bloc resection of urachus and umbilicus, pelvic lymphadenectomy, and ileocystoplasty. There were no complications seen in this patient. Neither urachal adenocarcinoma recurrence, metastasis, nor de novo uroileal cancer developed during 48-month follow-up. His reconstructed bladder functioned efficiently, without compromising the transplanted kidney function. Our case demonstrated that conservative surgery and augmentation ileocystoplasty could be offered to kidney transplant recipients with localized urachal carcinoma. PMID- 19545761 TI - Differentiation between infection in kidney and liver cysts in autosomal dominant polycystic kidney disease: use of PET-CT in diagnosis and to guide management. AB - Positron emission tomography-computed tomography (PET-CT) is a useful imaging method for localizing infective lesions. We report a case of autosomal dominant polycystic kidney disease in which PET-CT was used to differentiate between infection in the kidney and liver cysts. Localization of infection to the liver rather than to kidney cysts altered patient management. We briefly review the role of PET-CT in localization of an occult focus of infection. PMID- 19545762 TI - Tumor lysis syndrome occurring after the administration of rituximab for posttransplant lymphoproliferative disorder. AB - Tumor lysis syndrome (TLS) may occur after the administration of rituximab for lymphoproliferative disorders. We describe the case of a heart transplant recipient who developed TLS after a single dose of rituximab for the treatment of posttransplant lymphoproliferative disorder. Because rituximab is being used more frequently, it is important for transplant physicians to be aware of this potential complication particularly after administering the first dose. PMID- 19545763 TI - Overlooked constrictive pericarditis as a cause of relapsing ascites and impairment of renal allograft function. AB - Herein we have presented a case of overlooked relapsing constrictive pericarditis with ascites in a renal transplant patient. The first symptoms of constriction appeared during hemodialysis. After a 2-year period of remission, they relapsed at 1 year after transplantation causing deterioration of renal function. We noted that constrictive pericarditis can be misdiagnosed due to symptoms characteristic of heart failure. The diagnosis should be evident from right heart catheterization and computed tomographic (CT) imaging of the thickness or calcification of the pericardium. Once the diagnosis was established surgical treatment allowed total recovery with improved graft function. PMID- 19545764 TI - Successfully rescued renal graft artery thrombosis by ex vivo thrombectomy: a case report. AB - A 40-year-old woman who had been suffering from type II diabetes mellitus and consequent end-stage renal disease underwent living related kidney transplantation. The graft renal artery was anastomosed to the right internal iliac artery (end-to-end). Postoperative renoscintigraphy demonstrated normal graft perfusion. The serum lactate dehydrogenase level increased abruptly at postoperative day 15 and digital subtraction angiography disclosed graft artery thrombosis. Despite an intervention using a metallic coil stent, the rapid formation of thrombus occluded the graft artery completely. In an emergent surgical operation, the graft was nephrectomized carefully and irrigated after extensive thrombectomy. The graft was reimplanted by using an internal iliac artery graft. After three consecutive hemodialysis treatments, the patient's kidney graft functioned well. She has been in good health with stable graft function for 3 years after the operation. PMID- 19545765 TI - Sweet syndrome after kidney transplantation. AB - The association between Sweet syndrome (acute febrile neutrophilic dermatosis) and malignancies, infection, and drugs has been well established, but the disorder has never been reported in a solid organ transplant recipient. We have presented the first reported case of Sweet syndrome connected with solid organ transplant. Our patient is a 38-year-old man who underwent deceased donor kidney transplant for focal and segmental glomerulosclerosis and after resuming dialysis 6 weeks posttransplant, was readmitted 2 months later with high fevers and multiple head, neck, chest, and back lesions. Cultures were negative, and skin biopsy was consistent with Sweet syndrome. The lesions responded to higher doses of prednisone. Sweet syndrome has been linked to multiple drugs and malignancies, but has also been linked with states of altered immunity. Posttransplant immunosuppression may be related to this occurrence. PMID- 19545766 TI - Posterior reversible encephalopathy syndrome posttransplantation: a case report of possible association with cerebrospinal fluid leak after epidural catheterization. AB - We have described a case of posterior reversible encephalopathy syndrome (PRES) that developed early in the course of a renal transplant recipient. The patient was on cyclosporine and also had a cerebrospinal fluid (CSF) leak due to migration of an epidural catheter. The possibilities of PRES being associated with administration of cyclosporine and development of a CSF leak are discussed. PMID- 19545767 TI - Is localized prostate cancer an obstacle for an immediate consideration for renal transplantation? A case report. AB - Currently, renal failure patients with a history of prostate cancer are recommended to have a 2 to 5-year disease-free interval prior to being allowed to receive a kidney transplant. This disease is now amenable to curative therapy if diagnosed at an early stage when the tumor is organ-confined. We report a patient undergoing immediate renal transplantation following a laparoscopic radical prostatectomy for the treatment of prostate cancer. Candidates for renal transplantation who are diagnosed with early stage, organ-confined prostate cancer may be immediately considered for transplantation following radical prostatectomy in view of the high likelihood of cure of their prostate cancer. PMID- 19545768 TI - The superficial femoral vein: a valuable conduit for a short renal vein in kidney transplantation. AB - A variety of techniques have been developed to improve the problem with a short renal vein in kidney transplantation. Those techniques range from thorough mobilization of the recipient common and external iliac veins (iliac vein transposition) to donor vein elongation with a saphenous or gonadal vein or a polytetrafluoroethylene graft. Right renal vein extension using the inferior vena cava represents an excellent option for cadaveric kidney transplantation; however, for kidneys from living donors, that is not a suitable alternative. We present two cases where the superficial femoral vein was used as a conduit with good results; there was no additional morbidity for the patient. Although uncommon, renal vein extensions are sometimes needed in kidney transplantation. The superficial femoral vein comes close to representing the ideal conduit for a short renal vein when compared with a synthetic or an autologous graft due to its diameter and harvesting technique. PMID- 19545769 TI - Posttransplant lymphoproliferative disorder in the wall of a lymphocele: a case report. AB - Posttransplant lymphoproliferative disorder (PTLD) is a well-known complication of renal transplantation with increased incidence after introduction of more powerful immunosuppressive drugs. Presenting symptoms are nonspecific; some patients may be entirely asymptomatic. Herein we have reported a case of PTLD arising in the lymphocele wall presenting with B-symptoms and deterioration of graft function. A 62-year-old-female with end-stage renal disease secondary to Balkan endemic nephropathy and positive Epstein-Barr virus (EBV) serology before transplantation received a renal transplant from a deceased donor. Six months after transplantation she was admitted to the hospital with a 1-week history of malaise, weight loss, anorexia, night sweats, and febrile episodes. Multisliced computed tomography demonstrated a cystic structure at the renal hilus. Graft function deteriorated, so the patient underwent puncture of the lymphocele. Urgent graftectomy was necessary to stop the bleeding. Pathohistology demonstrated EBV-positive, CD20-positive PTLD. The patient received 6 cycles of chemotherapy and continued on hemodialysis. We concluded that a high index of suspicion for PTLD should be maintained when evaluating lymphoceles arising in the later posttransplantation period. Irrespective of their imaging features, biopsy should be performed to exclude PTLD. PMID- 19545770 TI - Portopulmonary hypertension: imatinib as a novel treatment and the Emory experience with this condition. AB - Portopulmonary hypertension (PoPH) is a common and feared complication of end stage liver disease, and imposes increased risk of perioperative morbidity in the liver transplant patient. Herein, we present the first successful use of Imatinib in the perioperative management of a patient who was not responding to conventional treatments as well as our institution's experience with this devastating complication. Of patients evaluated for transplant, 4.1% were identified with PoPH, half of which were listed, and one quarter of which were transplanted. Patients with PoPH were twice as likely to be transplanted than all other candidates (48% vs 25%), though less likely to survive their first year (69% vs 86.4%). PMID- 19545771 TI - Breakthrough rhinocerebral mucormycosis in a liver transplant patient receiving caspofungin. AB - Zygomycetes are among the most frequent causes of non-Aspergillus mycelial fungal infections in transplant recipients. We have described a single case of breakthrough zygomycosis. A young Japanese woman presented because of idiopathic fulminant hepatitis and renal failure. On the third day of admission, she underwent orthotopic liver transplantation. A considerable amount of red blood cells and fresh frozen plasma were transfused during surgery. On posttransplant day 2, Candida albicans was isolated from respiratory secretions; prophylactic caspofungin was prescribed. During the next 6 days, C albicans was isolated from tracheal secretions, surgical wound, and exudates and stools. Ventilator associated pneumonia was diagnosed day 4. Her renal function did not improve during the postoperative period; the patient continued on hemodialysis. On day 28, a dark blue eschar due to zygomycosis was detected on the skin of the nose. Tracheal and nasal exudates yielded Rhizopus sp. The patient died 12 hours later due to multiorgan failure with hypothermia. The fatal evolution in this case may be related to a presumed brain infarction after progressive vessel fungal invasion. The presented case had 2 risk factors related to zygomycosis. A high index of suspicion is required in transplant recipients with risk factors for zygomycosis. Early diagnosis and surgery with appropriate systemic fungal drugs (amphotericin B) are mandatory to improve the prognosis. PMID- 19545772 TI - Rapid normalization of portopulmonary hypertension after living donor liver transplantation. AB - Portopulmonary hypertension (PPHTN) is a relatively rare complication of end stage liver disease, and a serious problem in the context of liver transplantation. Herein we have reported a case of decompensated liver cirrhosis with PPHTN, which rapidly resolved after adult-to-adult living donor liver transplantation (LDLT). A 54-year-old man was referred to our hospital with end stage liver cirrhosis owing to chronic hepatitis C. Preoperative mean pulmonary artery pressure (mPAP), as assessed by right heart catheterization, was 38 mm Hg. Continuous infusion of epoprostenol decreased the mPAP to 24 mm Hg over 44 days. He underwent LDLT using a right hepatic lobe graft donated by his son. The postoperative course was uneventful, epoprostenol was weaned by postoperative day (POD) 21, and the mPAP normalized to 21 mm Hg on POD 28. The patient was discharged on POD 31 without any vasodilators. Our case revealed that liver transplantation can rapidly resolve PPTHN. PMID- 19545773 TI - Aggravation of mitral regurgitation by calcium administration in a patient with hypertrophic cardiomyopathy during liver transplantation: a case report. AB - Aggravation of mitral regurgitation (MR) due to left ventricular outflow tract obstruction (LVOTO) is likely to occur during liver transplantation in cirrhotic patients with hypertrophic cardiomyopathy (HCMP). Moreover, calcium administration following severe hypocalcemia due to inadequate citrate metabolism and massive transfusion may induce MR aggravation with LVOTO in such patients. Herein we have described a cirrhotic patient with HCMP in whom MR was aggravated due to LVOTO resulting from inadvertent rapid administration of calcium during liver transplantation. PMID- 19545774 TI - Complete neurological recovery from fulminant hepatic failure with subarachnoid hemorrhage by living donor liver transplantation: a case report. AB - A 29-year-old man was referred to our hospital with fulminant hepatic failure (FHF) and stage III hepatic coma (somnolence and confusion). Living donor liver transplantation (LDLT) was planned for 2 days after admission to our hospital. However, on the day after admission, he lapsed into stage IV hepatic coma: no right reflexes and no response to pain stimuli. Emergency cranial computed tomography revealed a subarachnoid hemorrhage (SAH), but no aneurysm was seen on magnetic resonance angiography. We speculated that the cause of the SAH may have been bleeding of intracranial veins secondary to coagulopathy and overextension of a vein due to brain edema. We considered that only LDLT could improve the coagulopathy and brain edema. The patient recovered consciousness on postoperative day (POD) 2 and was finally discharged from the hospital without neurological deficit on POD 85. This case suggested that SAH is not a prohibiting factor for LDLT in an FHF patient if the cause of the SAH is venous bleeding. PMID- 19545776 TI - Splenic artery steal syndrome after liver transplantation: an alternative technique of embolization. AB - Arterial steal syndrome after orthotopic liver transplantation (OLT) is characterized by arterial hypoperfusion of the graft, which is caused by a shift in blood flow into the splenic or gastroduodenal arteries. It causes hepatic hypoperfusion with attendant clinical manifestations of elevated liver function enzymes, allograft dysfunction, and cholestasis. Left untreated, the condition has a significant potential risk for postoperative morbidity and graft loss. Herein we have reported the case of a 68-year-old woman who developed splenic artery steal syndrome (SASS) after deceased donor liver transplantation. She was diagnosed by duplex Doppler ultrasonography and celiac trunk angiography, and subsequently treated with splenic artery embolization. PMID- 19545775 TI - The role of liver transplantation in the treatment of hepatic myelopathy: case report with review of the literature. AB - BACKGROUND AND AIMS: Hepatic myelopathy (HM) is a rare neurological complication of chronic liver disease, causing progressive spastic paraparesis. Unlike hepatic encephalopathy, the conservative treatment of HM is usually considered inefficient. Although some scattered case studies have reported that the clinical symptoms and signs of patients with HM were improved after orthotopic liver transplantation (OLT), there have also been cases of failure. The aim of this study was to identify the role of OLT in the treatment of HM. PATIENTS AND METHODS: We present two patients, who underwent OLT at the early stage after progressive HM had been diagnosed, gaining definite improvement of their neurological status within a relatively short time. In addition, we review all five similar cases that have hitherto been reported in English literature. RESULTS: The clinical symptoms and signs of HM improved to variable degrees after OLT in most cases except one with a long HM history. The degree of improvement correlated with the time interval between the onset of the first symptoms of HM and OLT. CONCLUSIONS: OLT represents an effective therapeutic option for HM. The duration of clinical manifestations of HM before OLT may be the main factor affecting the therapeutic outcome. PMID- 19545777 TI - Hepatic resection after liver transplantation as a graft-saving procedure. AB - Biliary lesions and hepatic artery thrombosis are known causes of posttransplant liver failure and liver retransplantation. The shortage of organs and the results of retransplantation have forced transplant teams to developed graft-saving techniques. We report two cases who underwent hepatic resection after liver transplantation. In both cases, a left lateral segmentectomy was performed. At follow-up, the patients are well with optimal graft function. We believe this kind of resection represents an adequate alternative in selected cases and must be considered before enlistment for retransplantation. PMID- 19545778 TI - Infrahepatic terminolateral cavocavostomy: a case report. AB - Infrahepatic vena cavocavostomy has been reported to be a rescue therapy when venous outflow from a liver allograft is obstructed due to stenosis of a piggyback anastomosis. The authors have described herein two consecutive adult liver transplantations using this technique as the primary venous anastomosis. Using a caval clamp positioned above the retrohepatic portion, partial hemodynamic obstruction of caval flow was well tolerated, avoiding use of a venovenous bypass. Although additional studies regarding this technique are needed, we believe that an infrahepatic vena cavocavostomy should be considered to be an alternative technique in carefully selected cases. PMID- 19545779 TI - Postshunt hemochromatosis leading to cardiogenic shock in a patient presenting for orthotopic liver transplant: a case report. AB - INTRODUCTION: In this case report, we have presented a patient whose liver transplant course was greatly affected by a previously undiagnosed disease process that ultimately led to an unexpected perioperative death. CASE REPORT: A 52-year-old woman with idiopathic hepatoportal sclerosis presented for liver transplantation 2 years postmesocaval shunt placement. Lab and pathology studies at the time of liver biopsy and shunt placement were negative for iron deposition. Preoperative workup was negative for cardiac disease. At the outset of her liver transplant, the patient developed refractory hypotension secondary to cardiogenic shock. Intraoperative transesophageal echocardiography and postoperative transthoracic echocardiography demonstrated dilated cardiomyopathy with severely depressed systolic function. Upon succumbing to ventricular fibrillation cardiac arrest, an autopsy revealed hemochromatosis of the heart, pancreas, kidneys, adrenals, and explanted liver. CONCLUSION: Dilated cardiomyopathy, congestive heart failure, and other unexpected disease processes resulting from hemochromatosis can greatly influence the care of postshunt liver failure patients. PMID- 19545780 TI - Can liver transplantation achieve similar effects at high altitudes compared with plains: case report. AB - In orthotopic liver transplantation, particular emphasis must be placed on the unique physiologic, pathologic, and clinical features in residents living in areas at high vs low altitude. Hypobaric hypoxia, hypothermia, heavy radiation, high wind speed, and superevaporation at high altitudes may lead to various diseases. These features have progressive effects on cardiopulmonary and central nervous system functions. A high concentration of red cells in the circulation is likely to result in an increased incidence of hepatic artery and portal vein thrombosis. The immune system is also affected at high altitudes. Exposure to high altitude, which is associated with decreased oxygen pressure, can result in oxidation-reduction stress, enhanced generation of reactive oxygen and nitrogen species, and related oxidative damage to lipids, proteins, and DNA. Our male patient with liver cirrhosis caused by chronic hepatitis B virus infectionunderwent orthotopic liver transplantation in Tibet with a successful outcome and good long-term survival. PMID- 19545782 TI - Accelerated partial breast irradiation: caution and concern from an ASTRO task force. PMID- 19545781 TI - Two different putative genetic animal models of childhood depression--a review. AB - Estimated prevalence of childhood and adolescent depression varies from 0.4% to 3% at the 0-12 age range and 3.3% to 12.4% at the 13-18 age range. Despite similarities in the clinical picture of major depression in children, adolescents, and adults, there are notable differences in the neurobiological correlates and treatment response of depressed patients in these different age cohorts. In contrast to adults, most depressed children fail to respond to antidepressants. The main aim of this paper is to review several studies which attempt to develop and examine genetic animal models for childhood depression, in order to enable a search for new, unique treatment approaches for depressed children. Two different "depressive-like" rat strains were studied: Flinders Sensitive Line (FSL) and their controls, Sprague-Dawley (SD) rats; and the Wistar Kyoto (WKY) strain and their controls, Wistar rats. The results suggest that prepubertal FSL and WKY rats exhibit different styles of depressive behavior, one co-morbid with "anxiety-like" behavior (WKY), and one that is not (FSL). These two profiles may model clinical characteristics that resemble two subgroups of depressed children. However, in general the data on the WKY rats would seem most consistent with a classic childhood depressive profile. The FSL profile may possibly be related to chronic stress, and its role as a potential model of childhood depression requires further support. These two different putative genetic animal models of childhood depression can help in the attempts to understand the neurobiological basis and to predict successful treatment strategies for different patterns of childhood psychopathology. PMID- 19545783 TI - The most important question. PMID- 19545784 TI - Accelerated partial breast irradiation consensus statement from the American Society for Radiation Oncology (ASTRO). AB - PURPOSE: To present guidance for patients and physicians regarding the use of accelerated partial-breast irradiation (APBI), based on current published evidence complemented by expert opinion. METHODS AND MATERIALS: A systematic search of the National Library of Medicine's PubMed database yielded 645 candidate original research articles potentially applicable to APBI. Of these, 4 randomized trials and 38 prospective single-arm studies were identified. A Task Force composed of all authors synthesized the published evidence and, through a series of meetings, reached consensus regarding the recommendations contained herein. RESULTS: The Task Force proposed three patient groups: (1) a "suitable" group, for whom APBI outside of a clinical trial is acceptable, (2) a "cautionary" group, for whom caution and concern should be applied when considering APBI outside of a clinical trial, and (3) an "unsuitable" group, for whom APBI outside of a clinical trial is not generally considered warranted. Patients who choose treatment with APBI should be informed that whole-breast irradiation (WBI) is an established treatment with a much longer track record that has documented long-term effectiveness and safety. CONCLUSION: Accelerated partial-breast irradiation is a new technology that may ultimately demonstrate long-term effectiveness and safety comparable to that of WBI for selected patients with early breast cancer. This consensus statement is intended to provide guidance regarding the use of APBI outside of a clinical trial and to serve as a framework to promote additional clinical investigations into the optimal role of APBI in the treatment of breast cancer. PMID- 19545785 TI - Bath and shower effects in the rat parotid gland explain increased relative risk of parotid gland dysfunction after intensity-modulated radiotherapy. AB - PURPOSE: To assess in a rat model whether adding a subtolerance dose in a region adjacent to a high-dose irradiated subvolume of the parotid gland influences its response (bath-and-shower effect). METHODS AND MATERIALS: Irradiation of the whole, cranial 50%, and/or the caudal 50% of the parotid glands of Wistar rats was performed using 150-MeV protons. To determine suitable (i.e., subtolerance) dose levels for a bath-dose, both whole parotid glands were irradiated with 5 to 25 Gy. Subsequently groups of Wistar rats received 30 Gy to the caudal 50% (shower) and 0 to 10 Gy to the cranial 50% (bath) of both parotid glands. Stimulated saliva flow rate (function) was measured before and up to 240 days after irradiation. RESULTS: Irradiation of both glands up to a dose of 10 Gy did not result in late loss of function and is thus regarded subtolerance. Addition of a dose bath of 1 to 10 Gy to a high-dose in the caudal 50% of the glands resulted in enhanced function loss. CONCLUSION: Similar to the spinal cord, the parotid gland demonstrates a bath and shower effect, which may explain the less than-expected sparing of function after IMRT. PMID- 19545786 TI - Efficacy of sunitinib and radiotherapy in genetically engineered mouse model of soft-tissue sarcoma. AB - PURPOSE: Sunitinib (SU) is a multitargeted receptor tyrosine kinase inhibitor of the vascular endothelial growth factor and platelet-derived growth factor receptors. The present study examined SU and radiotherapy (RT) in a genetically engineered mouse model of soft tissue sarcoma (STS). METHODS AND MATERIALS: Primary extremity STSs were generated in genetically engineered mice. The mice were randomized to treatment with SU, RT (10 Gy x 2), or both (SU+RT). Changes in the tumor vasculature before and after treatment were assessed in vivo using fluorescence-mediated tomography. The control and treated tumors were harvested and extensively analyzed. RESULTS: The mean fluorescence in the tumors was not decreased by RT but decreased 38-44% in tumors treated with SU or SU+RT. The control tumors grew to a mean of 1378 mm(3) after 12 days. SU alone or RT alone delayed tumor growth by 56% and 41%, respectively, but maximal growth inhibition (71%) was observed with the combination therapy. SU target effects were confirmed by loss of target receptor phosphorylation and alterations in SU-related gene expression. Cancer cell proliferation was decreased and apoptosis increased in the SU and RT groups, with a synergistic effect on apoptosis observed in the SU+RT group. RT had a minimal effect on the tumor microvessel density and endothelial cell-specific apoptosis, but SU alone or SU+RT decreased the microvessel density by >66% and induced significant endothelial cell apoptosis. CONCLUSION: SU inhibited STS growth by effects on both cancer cells and tumor vasculature. SU also augmented the efficacy of RT, suggesting that this combination strategy could improve local control of STS. PMID- 19545788 TI - Induction and rejoining of DNA double strand breaks assessed by H2AX phosphorylation in melanoma cells irradiated with proton and lithium beams. AB - PURPOSE: The aim of this study was to evaluate the induction and rejoining of DNA double strand breaks (DSBs) in melanoma cells exposed to low and high linear energy transfer (LET) radiation. METHODS AND MATERIALS: DSBs and survival were determined as a function of dose in melanoma cells (B16-F0) irradiated with monoenergetic proton and lithium beams and with a gamma source. Survival curves were obtained by clonogenic assay and fitted to the linear-quadratic model. DSBs were evaluated by the detection of phosphorylated histone H2AX (gammaH2AX) foci at 30 min and 6 h post-irradiation. RESULTS: Survival curves showed the increasing effectiveness of radiation as a function of LET. gammaH2AX labeling showed an increase in the number of foci vs. dose for all the radiations evaluated. A decrease in the number of foci was found at 6 h post-irradiation for low LET radiation, revealing the repair capacity of DSBs. An increase in the size of gammaH2AX foci in cells irradiated with lithium beams was found, as compared with gamma and proton irradiations, which could be attributed to the clusters of DSBs induced by high LET radiation. Foci size increased at 6 h post-irradiation for lithium and proton irradiations in relation with persistent DSBs, showing a correlation with surviving fraction. CONCLUSIONS: Our results showed the response of B16-F0 cells to charged particle beams evaluated by the detection of gammaH2AX foci. We conclude that gammaH2AX foci size is an accurate parameter to correlate the rejoining of DSBs induced by different LET radiations and radiosensitivity. PMID- 19545789 TI - S-phase cells are more sensitive to high-linear energy transfer radiation. AB - PURPOSE: S-phase cells are more resistant to low-linear energy transfer (LET) ionizing radiation (IR) than nonsynchronized and G(1)-phase cells, because both nonhomologous end-joining (NHEJ) and homologous recombination repair can repair DNA double-strand breaks (DSBs) in the S phase. Although it was reported 3 decades ago that S-phase cells did not show more resistance to high-LET IR than cells in other phases, the mechanism remains unclear. We therefore attempted to study the phenotypes and elucidate the mechanism involved. METHODS AND MATERIALS: Wild-type and NHEJ-deficient cell lines were synchronized using the double thymidine approach. A clonogenic assay was used to detect the sensitivity of nonsynchronized, synchronized S-phase, and G(2)-phase cells to high- and low-LET IR. The amounts of Ku bound to DSBs in the high- and low-LET-irradiated cells were also examined. RESULTS: S-phase wild-type cells (but not NHEJ-deficient cells) were more sensitive to high-LET IR than nonsynchronized and G(2)-phase cells. In addition, S-phase wild-type cells showed less efficient Ku protein binding to DSBs than nonsynchronized and G(2)-phase cells in response to high-LET IR, although all cells at all phases showed similarly efficient levels of Ku protein binding to DSBs in response to low-LET IR. CONCLUSIONS: S-phase cells are more sensitive to high-LET IR than nonsynchronized and G(2)-phase cells, because of the following mechanism: it is more difficult for Ku protein to bind to high LET IR-induced DNA DSBs in S-phase cells than in cells at other phases, which results in less efficient NHEJ. PMID- 19545787 TI - Celastrol potentiates radiotherapy by impairment of DNA damage processing in human prostate cancer. AB - PURPOSE: Celastrol is an active ingredient of traditional herbal medicine and has recently been identified as a potent natural proteasome inhibitor. In the present study, we evaluated the radiosensitizing potential of celastrol in the human prostate cancer PC-3 model. METHODS AND MATERIALS: Clonogenic assays were performed to determine the radiosensitizing effect of celastrol. Apoptosis was examined by flow cytometry using Annexin V and propidium iodide staining and by a caspase-3 activation assay. DNA damage processing was examined by immunofluorescent staining and Western blot for phosphorylated H2AX (gammaH2AX). The PC-3 xenograft model in the athymic nude mouse was used for the determination of the in vivo efficacy of celastrol combined with radiotherapy. The tumor samples were also analyzed for apoptosis and angiogenesis. RESULTS: Celastrol sensitized PC-3 cells to ionizing radiation (IR) in a dose- and schedule dependent manner, in which pretreatment with celastrol for 1 h followed by IR achieved maximal radiosensitization. Celastrol significantly prolonged the presence of IR-induced gammaH2AX and increased IR-induced apoptosis. Celastrol, combined with fractionated radiation, significantly inhibited PC-3 tumor growth in vivo without obvious systemic toxicity. The combination treatment increased gammaH2AX levels and apoptosis, induced cleavage of poly(adenosine diphosphate ribose)polymerase and Mcl-1, and reduced angiogenesis in vivo compared with either treatment alone. CONCLUSION: Celastrol sensitized PC-3 cells to radiation both in vitro and in vivo by impairing DNA damage processing and augmenting apoptosis. Celastrol might represent a promising new adjuvant regimen for the treatment of hormone-refractory prostate cancer. PMID- 19545790 TI - Differential effect triggered by a heparan mimetic of the RGTA family preventing oral mucositis without tumor protection. AB - PURPOSE: Oral mucositis is a common side effect induced by radio/chemotherapy in patients with head and neck cancer. Although it dramatically impairs patient quality of life, no efficient and safe therapeutic solution is available today. Therefore, we investigated the protective efficacy of a new heparan mimetic biopolymer, RGTA-OTR4131, used alone or in combination with amifostine, for oral mucositis and simultaneously evaluated its effect on tumor growth in vitro and in vivo. METHODS AND MATERIALS: A single dose of 16.5 Gy was selectively delivered to the snout of mice, and the effects of OTR4131 or amifostine-OTR4131 were analyzed by macroscopic scoring and histology. The effect of OTR4131 administration on tumor growth was then investigated in vitro and in xenograft models using two cell lines (HEP-2 and HT-29). RESULTS: Amifostine and OTR4131 significantly decreased the severity and duration of lip mucosal reactions. However, amifostine has to be administered before irradiation, whereas the most impressive protection was obtained when OTR4131 was injected 24 h after irradiation. In addition, OTR4131 was well tolerated, and the combination of amifostine and OTR4131 further enhanced mucosal protection. At the tumor level, OTR4131 did not modify HEP-2 cell line clonogenic survival in vitro or protect xenografted tumor cells from radiotherapy. Of interest, high doses of OTR4131 significantly decreased clonogenic survival of HT-29 cells. CONCLUSIONS: RGTAs OTR4131 is a well-tolerated, natural agent that effectively reduces radio-induced mucositis without affecting tumor sensitivity to irradiation. This suggests a possible transfer into the clinic for patients' benefit. PMID- 19545791 TI - Rad51 protein expression and survival in patients with glioblastoma multiforme. AB - PURPOSE: Treatment of glioblastoma multiforme (GBM) continues to pose a significant therapeutic challenge, with most tumors recurring within the previously irradiated tumor bed. To improve outcomes, we must be able to identify and treat resistant cell populations. Rad51, an enzyme involved in homologous recombinational repair, leads to increased resistance of tumor cells to cytotoxic treatments such as radiotherapy. We hypothesized that Rad51 might contribute to GBM's apparent radioresistance and consequently influence survival. METHODS AND MATERIALS: A total of 68 patients with an initial diagnosis of GBM were retrospectively evaluated; for 10 of these patients, recurrent tumor specimens were used to construct a tissue microarray. Rad51 protein expression was then correlated with the actual and predicted survival using recursive partitioning analysis. RESULTS: Rad51 protein was elevated in 53% of the GBM specimens at surgery. The Rad51 levels correlated directly with survival, with a median survival of 15 months for patients with elevated Rad51 compared with 9 months for patients with low or absent levels of Rad51 (p = .05). At disease recurrence, 70% of patients had additional increases in Rad51 protein. Increased Rad51 levels at disease recurrence similarly predicted for improved overall survival, with a mean survival of 16 months from the second craniotomy compared with only 4 months for patients with low Rad51 levels (p = .13). CONCLUSION: Elevated levels of the double-stranded DNA repair protein Rad51 predicted for an increase survival duration in patients with GBM, at both initial tumor presentation and disease recurrence. PMID- 19545792 TI - Tissue feature-based and segmented deformable image registration for improved modeling of shear movement of lungs. AB - PURPOSE: To report a tissue feature-based image registration strategy with explicit inclusion of the differential motions of thoracic structures. METHODS AND MATERIALS: The proposed technique started with auto-identification of a number of corresponding points with distinct tissue features. The tissue feature points were found by using the scale-invariant feature transform method. The control point pairs were then sorted into different "colors" according to the organs in which they resided and used to model the involved organs individually. A thin-plate spline method was used to register a structure characterized by the control points with a given "color." The proposed technique was applied to study a digital phantom case and 3 lung and 3 liver cancer patients. RESULTS: For the phantom case, a comparison with the conventional thin-plate spline method showed that the registration accuracy was markedly improved when the differential motions of the lung and chest wall were taken into account. On average, the registration error and standard deviation of the 15 points against the known ground truth were reduced from 3.0 to 0.5 mm and from 1.5 to 0.2 mm, respectively, when the new method was used. A similar level of improvement was achieved for the clinical cases. CONCLUSION: The results of our study have shown that the segmented deformable approach provides a natural and logical solution to model the discontinuous organ motions and greatly improves the accuracy and robustness of deformable registration. PMID- 19545793 TI - Effects of respiration-induced density variations on dose distributions in radiotherapy of lung cancer. AB - PURPOSE: To determine the effect of respiration-induced density variations on the estimated dose delivered to moving structures and, consequently, to evaluate the necessity of using full four-dimensional (4D) treatment plan optimization. METHODS AND MATERIALS: In 10 patients with large tumor motion (median, 1.9 cm; range, 1.1-3.6 cm), the clinical treatment plan, designed using the mid ventilation ([MidV]; i.e., the 4D-CT frame closest to the time-averaged mean position) CT scan, was recalculated on all 4D-CT frames. The cumulative dose was determined by transforming the doses in all breathing phases to the MidV geometry using deformable registration and then averaging the results. To determine the effect of density variations, this cumulative dose was compared with the accumulated dose after similarly deforming the planned (3D) MidV-dose in each respiratory phase using the same transformation (i.e., "blurring the dose"). RESULTS: The accumulated tumor doses, including and excluding density variations, were almost identical. Relative differences in the minimum gross tumor volume (GTV) dose were less than 2% for all patients. The relative differences were even smaller in the mean lung dose and the V20 (<0.5% and 1%, respectively). CONCLUSIONS: The effect of respiration-induced density variations on the dose accumulated over the respiratory cycle was very small, even in the presence of considerable respiratory motion. A full 4D-dose calculation for treatment planning that takes into account such density variations is therefore not required. Planning using the MidV-CT derived from 4D-CT with an appropriate margin for geometric uncertainties is an accurate and safe method to account for respiration-induced anatomy variations. PMID- 19545794 TI - A novel device for intravaginal electronic brachytherapy. AB - PURPOSE: Postoperative intravaginal brachytherapy for endometrial carcinoma is usually performed with (192)Ir high-dose rate (HDR) afterloading. A potential alternative is treatment with a broadband 50kV X-ray point source, the advantage being its low energy and the consequential steep dose gradient. The aim of this study was to create and evaluate a homogeneous cylindrical energy deposition around a newly designed vaginal applicator. METHODS AND MATERIALS: To create constant isodose layers along the cylindrical plastic vaginal applicator, the source (INTRABEAM system) was moved in steps of 17-19.5 mm outward from the tip of the applicator. Irradiation for a predetermined time was performed at each position. The axial shift was established by a stepping mechanism that was mounted on a table support. The total dose/dose distribution was determined using film dosimetry (Gafchromic EBT) in a "solid water" phantom. The films were evaluated with Mathematica 5.2 and OmniPro-I'mRT 1.6. The results (dose D0/D5/D10 in 0/5/10 mm tissue depth) were compared with an (192)Ir HDR afterloading plan for multiple sampling points around the applicator. RESULTS: Three different dose distributions with lengths of 3.9-7.3 cm were created. The irradiation time based on the delivery of 5/7 Gy to a 5 mm tissue depth was 19/26 min to 27/38 min. D0/D5/D10 was 150%/100%/67% for electronic brachytherapy and 140%/100%/74% for the afterloading technique. The deviation for repeated measurements in the phantom was <7%. CONCLUSIONS: It is possible to create a homogeneous cylindrical dose distribution, similar to (192)Ir HDR afterloading, through the superimposition of multiple spherical dose distributions by stepping a kilovolt point source. PMID- 19545795 TI - Pattern analysis of acute mucosal reactions in patients with head and neck cancer treated with conventional and accelerated irradiation. In regard to Wygodaa, et al. (Int J Radiat Oncol Biol Phys 2008;73:384-390). PMID- 19545798 TI - Outcome prediction after surgery and chemoradiation of head-and-neck squamous cell carcinoma (HNSCC), using baseline perfusion computed tomography (CT) microcirculatory parameters vs. tumor volume. PMID- 19545799 TI - New abbreviations for visual acuity values. PMID- 19545800 TI - Surgical technique for small-incision intraocular lens exchange. AB - We present a simple, effective technique to remove an intraocular lens (IOL) via a very small incision (2.2 mm). After the IOL is carefully removed from the capsular bag, into the anterior chamber, 2 radial incisions, 35 degrees to 50 degrees apart, are made with a Vannas scissors. This results in 2 IOL pieces (maximum width of 2.70 mm) that can be easily explanted through the unenlarged small clear corneal incisions (2.2 mm) in use today. PMID- 19545801 TI - Combined microwave energy and fixative agent for cataract induction in pig eyes. AB - We describe a technique used in enucleated pig eyes to create a uniformly hardened lens nucleus and an anterior capsule similar to the cataractous human lens. After treatment with microwave heat and a fixative agent, the pig eyes have clear corneas, anterior capsules with less tension and elasticity, and harder lens nuclei, with a consistency similar to that of a nuclear sclerosis grade I to V in the Emery-Little classification. The eyes can be used by residents in training to practice various phacoemulsification techniques, including continuous curvilinear capsulorhexis, hydrodelineation, sculpting, divide and conquer, phaco chop, and nondividing phacoemulsification. PMID- 19545803 TI - Posterior chamber phakic intraocular lenses after penetrating keratoplasty. AB - PURPOSE: To evaluate the efficacy, predictability, and safety of a phakic posterior chamber intraocular Collamer lens (ICL) after penetrating keratoplasty (PKP). SETTING: Fernandez-Vega Ophthalmological Institute, Oviedo, Spain. METHODS: A myopic or toric ICL was implanted after PKP in eyes unable to wear glasses or contact lenses and for which corneal laser surgery was contraindicated. The uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), refractive error, and endothelial cell count were recorded preoperatively and 24 months postoperatively. RESULTS: Preoperatively, the 15 eyes had myopia from -2.00 to -17.00 diopters (D) or astigmatism from 1.50 to -7.00 D. Twenty-four months postoperatively, the mean Snellen decimal UDVA was 0.51 +/- 0.30 (SD). The UDVA was 20/40 or better in 7 eyes (46.6%). The mean CDVA was 0.79 +/- 0.22. The CDVA was 20/40 or better in 12 eyes (80%) and 20/25 in 6 eyes (40%). No eye lost more than 1 line of acuity, 2 eyes gained 1 line, and 5 eyes gained more than 2 lines; 8 eyes were unchanged. The safety index was 1.58. The spherical equivalent (SE) was within +/-1.00 D in 80% of eyes and within +/-0.50 D in 66.6% of eyes. The mean postoperative SE was -0.95 +/- 1.12 D. At 24 months, the mean vault was 2.06 +/- 0.96 and the mean endothelial cell loss, 8.1%. CONCLUSION: Results indicate that phakic intraocular lens implantation is a viable treatment for myopia and astigmatism after PKP in patients for whom glasses, contact lenses, or corneal refractive surgery are contraindicated. PMID- 19545802 TI - One-year results of excimer laser in situ keratomileusis for hyperopia. AB - PURPOSE: To evaluate the visual and refractive results of hyperopic LASIK. SETTING: Clinica Baviera Instituto Oftalmologico Europeo, Madrid, Spain. METHODS: This retrospective consecutive noncomparative observational study evaluated hyperopic LASIK results over 1 year. Outcomes included uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), cycloplegic refraction, and corneal topography. Surgery was performed using an MEL 80-G excimer laser. Results were analyzed by preoperative spherical equivalent (SE) (Group 1: or=+3.60 D). RESULTS: The mean UDVA improved from 0.50 +/- 0.3 (SD) to 0.90 +/- 0.2 in Group 1 and from 0.50 +/- 0.3 to 0.80 +/- 0.2 in Group 2 and the mean CDVA, from 0.86 +/- 0.2 to 0.93 +/- 0.1 and from 0.80 +/- 0.2 to 0.90 +/- 0.2, respectively. The mean cycloplegic SE improved from +2. 5+/- 0.8 to +0.1 +/- 0.5 in Group 1 and from +4.5 +/- 0.6 to +0.4 +/- 0.6 in Group 2; 70.9% of eyes and 63.3% of eyes, respectively, were within +/-0.50 D of emmetropia. Postoperatively, 92.8% of eyes in Group 1 and 87.8% in Group 2 maintained or gained 1 or more lines of CDVA; 1.7% and 4.0%, respectively, lost 2 or more lines. The safety index was 1.1 in both groups and the efficacy index, 1.01 in Group 1 and 0.98 in Group 2. The enhancement rate was 20.0% and 18.4%, respectively. CONCLUSION: Excimer laser LASIK was safe and effective for treating hyperopia up to +6.25 D with no further loss of CDVA lines after enhancement. PMID- 19545804 TI - Back-calculation to model strategies for pretreatment adjustment of the ablation sphere in myopic wavefront laser in situ keratomileusis. AB - PURPOSE: To examine strategies for adjusting the ablation sphere in myopic wavefront laser in situ keratomileusis (LASIK) with reference to preoperative manifest refraction. SETTING: Refractive Surgery Service, Moorfields Eye Hospital, London, United Kingdom. METHODS: The variance in the 3-month postoperative manifest refraction spherical equivalent (MRSE) results in 295 consecutive cases of myopic wavefront LASIK treated with a nonsystematic, intuitive approach to pretreatment adjustment of the ablation sphere was compared with the variance in modeled results obtained by back-calculation using 4 systematic strategies: (1) no adjustment (No Adj), (2) addition of the difference between the preoperative MRSE and the preoperative 4.0 mm pupil wavefront refraction spherical equivalent (WRSE) (4.0 mm), (3) addition of the difference between the preoperative MRSE and the preoperative maximum pupil WRSE (Max), and (4) addition of the difference between the preoperative MRSE and the preoperative effective blur (EB). The EB is a theoretical value for the dioptric SE of all aberrations at the pupil size calculated for the Hartmann-Shack images acquired (wavefront diameter). A stratified analysis of results for different wavefront diameters was also performed. RESULTS: Variance in the postoperative MRSE was least (0.116) when nonsystematic, intuitive adjustments to the ablation sphere were used. In ascending order, the back-calculated variance was as follows: EB (0.142; P = .09, Bartlett test), 4.0 mm pupil (0.163; P = .004), No Adj (0.171; P = .001), and Max (0.225; P<.0001). There was a strong trend toward reduced variance in results in patients with a larger wavefront diameter. CONCLUSION: Back-calculation to model results with different pretreatment ablation adjustment strategies may be useful to eliminate unpromising new approaches before clinical trials. PMID- 19545805 TI - Accuracy of modern intraocular lens power calculation formulas in refractive lens exchange for high myopia and high hyperopia. AB - PURPOSE: To determine the accuracy of the Holladay 2, Hoffer Q, SRK/T, and Haigis intraocular lens (IOL) power calculation formulas in high myopic and high hyperopic refractive lens exchange (RLE) with hydrophobic acrylic foldable IOLs. SETTING: Department of Ophthalmology, Johann Wolfgang Goethe University, Frankfurt am Main, Germany. METHODS: Preoperatively, axial length (AL) was measured by partial coherence interferometry with the IOLMaster. The RLE was performed by phacoemulsification with posterior chamber IOL implantation in the capsular bag. Optimization of the lens constants for each IOL type was done separately for eyes with a positive lens and eyes with a negative lens. Prediction error (predicted refraction - postoperative refraction) and mean absolute error were back-calculated for the Holladay 2, Hoffer Q, SRK/T, and Haigis formulas. RESULTS: Sixty-three eyes (44 myopic, AL >or=26.0 mm; 19 hyperopic, AL 5.00 D) astigmatism. It aims also to show whether soft or rigid gas-permeable (GP) contact lenses offer the best correction and to determine which modality is preferred by subjects. METHODS: Twenty subjects were randomly fitted with both soft and GP lenses. Group A was first fitted with soft lenses then switched to GPs and vice versa for group B. For each type of lens worn, low- and high-contrast visual acuity (VA) and stereoscopy were evaluated at both near and far. Each subject was asked to select the lens type of choice and to rate quality of vision in day-to-day activities through a questionnaire. RESULTS: There was no significant difference in objective binocular VA between current spectacles and empirically calculated soft toric lenses and GP toric/bi-toric contact lenses at all distances. That was also true for stereoscopy. Subjectively, most of the subjects preferred rigid contact lenses because of the quality of their vision in day-to-day activities, and a majority of them decided to continue with that modality. CONCLUSION: Empirical fitting of toric soft or GP contact lenses leads to very good results, which suggests that each type of lens might constitute a viable option. PMID- 19545852 TI - Review and update: current treatment trends for patients with retinitis pigmentosa. AB - BACKGROUND: Retinitis pigmentosa (RP) is a heterogeneous group of inherited retinal disorders characterized by progressive photoreceptor apoptosis. It is the leading cause of inherited retinal degeneration-associated blindness. RP has a unique set of clinical characteristics that make it a complex disease associated with distinct inheritance patterns. An understanding of the pathogenesis is essential in the process of the differential diagnosis and the development of treatment options. Recent developments in research are likely to expand the various therapeutic modalities to include gene therapy, pharmacologic treatment, cell transplantation, and neuro-prosthetic devices. METHODS: A literature search was performed to comprehensively review RP diagnosis, pathophysiology, and treatment. CONCLUSION: Advances in the understanding of the pathophysiology of RP are creating new opportunities for the treatment of this often visually debilitating eye condition. Optometrists, as primary eye care practitioners, should be aware of the inheritance, pathophysiology, and current treatment options for RP as well as treatments in development so that they can best care for their patients with inherited retinal disorders. PMID- 19545853 TI - Eye fixation behaviour in the number bisection task: evidence for temporal specificity. AB - Together with magnitude representations, knowledge about multiplicativity and parity contributes to numerical problem solving. In the present study, we used eye tracking to document how and when multiplicativity and parity are recruited in the number bisection task. Fourteen healthy adults evaluated whether the central number of a triplet (e.g., 21_24_27) corresponds to the arithmetic integer mean of the interval defined by the two outer numbers. We observed multiplicativity to specifically affect gaze duration on numbers, indicating that the information of multiplicative relatedness is activated at early processing stages. In contrast, parity only affected total reading time, suggesting involvement in later processing stages. We conclude that different representational features of numbers are available and integrated at different processing stages within the same task and outline a processing model for these temporal dynamics of numerical cognition. PMID- 19545854 TI - Expression patterns of the Fam83h gene during murine tooth development. AB - AIM: Recently a novel gene, FAM83H, was identified by a genetic linkage study in the hypocalcified form of the amelogenesis imperfecta family with an autosomal dominant hereditary pattern. Little is known about this novel gene, and so we investigated the expression pattern of Fam83h in murine tooth development using serial sectional in situ hybridisation. METHODS AND MATERIALS: Using mandibles of ICR mouse at specific developmental stages, in situ hybridisation was performed by DIG-labeled RNA probe. RESULTS: Faint expression was detected in limited cells at embryonic day 14 (E14) in the molar. At the bell stage, E16, Fam83h was localised in the outer and inner enamel epithelium, as well as dental papilla. Fam83h expression begins on E15 in the developing incisor. At E18, Fam83h was expressed in the inner enamel epithelium of the apical bud, ameloblasts and odontoblasts. The expression was stronger in the presecretory stages than the secretory stages. CONCLUSION: Fam83h was detected in the ameloblasts from the presecretory to the secretory stage, and also the odontoblasts layer and surrounding alveolar bone. PMID- 19545855 TI - More lateral and anterior prefrontal coil location is associated with better repetitive transcranial magnetic stimulation antidepressant response. AB - BACKGROUND: The left dorsolateral prefrontal cortex (DLPFC) is the most commonly used target for transcranial magnetic stimulation (TMS) in the treatment of depression. The "5-cm rule" is an empiric method used for probabilistic targeting of the DLPFC in most clinical trials. This rule may be suboptimal, as it does not account for differences in skull size or variations in prefrontal anatomy relative to motor cortex location. This study is a post hoc analysis of data from a large repetitive TMS (rTMS) trial in which we examined the variability of coil placement and how it affects antidepressant efficacy. METHODS: Fifty-four depressed subjects enrolled in a randomized, single-site trial received either active rTMS or sham for 3 weeks. Prior to treatment initiation, investigators placed vitamin E capsules at the point of stimulation and used a high-resolution magnetic resonance imaging (MRI) scan to image these fiducials relative to anatomy. We employed a semiautomated imaging-processing algorithm to localize the cortical region stimulated. RESULTS: Active TMS significantly reduced Hamilton Depression Rating Scale (HDRS) scores. A linear model for this improvement involving the coordinates of the stimulated cortex location, age, and treatment condition was highly significant. Specifically, individuals with more anterior and lateral stimulation sites were more likely to respond. CONCLUSIONS: These results suggest that within the general anatomical area targeted by the 5-cm rule, placing the TMS coil more laterally and anteriorly is associated with improved response rates in TMS depression studies. Controlled studies testing this anatomical hypothesis are needed. PMID- 19545856 TI - Sensorimotor gating depends on polymorphisms of the serotonin-2A receptor and catechol-O-methyltransferase, but not on neuregulin-1 Arg38Gln genotype: a replication study. AB - BACKGROUND: Prepulse inhibition (PPI) of the acoustic startle response (ASR) is an operational measure of sensorimotor gating and a promising endophenotype of schizophrenia. We have recently shown that the linked serotonin-2A receptor (5 HT(2A)R) A-1438 G and T102C polymorphisms modulate PPI in schizophrenia patients. Moreover, it was shown that genetic variation in the catechol-O-methyltransferase (COMT) and the neuregulin-1 (NRG-1) proteins influences PPI in schizophrenia patients and healthy volunteers. Therefore, we aimed to replicate these results and investigated the impact of the related polymorphisms on PPI in healthy human volunteers. METHODS: We analyzed the 5-HT(2A)R A-1438 G/T102C (rs6311/rs6313), the COMT Val158Met (rs4680), and the NRG-1 Arg38Gln (rs3924999) polymorphisms, assessing startle reactivity, habituation, and PPI of ASR in 107 healthy Caucasian volunteers. RESULTS: Subjects homozygous for the 5-HT(2A)R T102C-T/A 1438 G-A allele showed increased PPI levels. In particular, male subjects with the COMT Met158Met-genotype also showed elevated PPI. The NRG-1 Arg38Gln genotype did not have a significant impact on PPI. Startle reactivity was not affected by any of the investigated polymorphisms. CONCLUSIONS: We confirmed in an independent sample of healthy volunteers that PPI is influenced by genetic variation in the 5-HT(2A)R gene. The influence of the COMT Val158Met genotype on PPI appears to be sex-specific. These results underscore the significance of the serotonin and dopamine systems in the modulation of sensorimotor gating. PMID- 19545857 TI - Effects of intravenous ketamine on explicit and implicit measures of suicidality in treatment-resistant depression. AB - BACKGROUND: Intravenous ketamine has shown rapid antidepressant effects in early trials, making it a potentially attractive candidate for depressed patients at imminent risk of suicide. The Implicit Association Test (IAT), a performance based measure of association between concepts, may have utility in suicide assessment. METHODS: Twenty-six patients with treatment-resistant depression were assessed using the suicidality item of the Montgomery-Asberg Depression Rating Scale (MADRS-SI) 2 hours before and 24 hours following a single subanesthetic dose of intravenous ketamine. Ten patients also completed IATs assessing implicit suicidal associations at comparable time points. In a second study, nine patients received thrice-weekly ketamine infusions over a 12-day period. RESULTS: Twenty four hours after a single infusion, MADRS-SI scores were reduced on average by 2.08 points on a 0 to 6 scale (p < .001; d = 1.37), and 81% of patients received a rating of 0 or 1 postinfusion. Implicit suicidal associations were also reduced following ketamine (p = .003; d = 1.36), with reductions correlated across implicit and explicit measures. MADRS-SI reductions were sustained for 12 days by repeated-dose ketamine (p < .001; d = 2.42). CONCLUSIONS: These preliminary findings support the premise that ketamine has rapid beneficial effects on suicidal cognition and warrants further study. PMID- 19545858 TI - Characterization of QKI gene expression, genetics, and epigenetics in suicide victims with major depressive disorder. AB - BACKGROUND: A number of studies have suggested deficits in myelination and glial gene expression in different psychiatric disorders. We examined the brain expression and genetic/epigenetic regulation of QKI, an oligodendrocyte-specific RNA binding protein important for cell development and myelination. METHODS: The microarray-based expression of QKI was evaluated in cortical and subcortical brain regions from suicide victims with a diagnosis of major depression (n = 16) and control subjects (n = 13). These findings were also assessed with a real-time (quantitative polymerase chain reaction [qPCR]) approach, with QKI protein levels evaluated through immunoblotting. Identification of a QKI promoter sequence was then used to examine genetic and epigenetic variation at the QKI locus. RESULTS: The messenger RNA (mRNA) levels of multiple transcripts of QKI were evaluated on Affymetrix microarrays, revealing significant reductions in 11 cortical regions and the hippocampus and amygdala of suicide victims compared with control subjects. Microarray findings were confirmed by qPCR, and reduced expression of QKI protein was identified in orbitofrontal cortex. Analysis of promoter variation and methylation state in a subset of individuals did not identify differences at the genetic or epigenetic level between depressed suicide victims and control subjects. CONCLUSIONS: The observation of consistent reductions in multiple isoforms of QKI mRNA in depressed suicide victims supports the growing body of evidence for a role of myelination-related deficits in the etiology of psychiatric disorders. A specific role of QKI in this process is implied by its reduced expression and known interactions with genes involved in oligodendrocyte determination; however, QKI gene variation responsible for these changes remains to be identified. PMID- 19545859 TI - Physiological responses to brain stimulation during limbic surgery: further evidence of anterior cingulate modulation of autonomic arousal. AB - BACKGROUND: In view of conflicting neuroimaging results regarding autonomic specific activity within the anterior cingulate cortex (ACC), we investigated autonomic responses to direct brain stimulation during stereotactic limbic surgery. METHODS: Skin conductance activity and accelerative heart rate responses to multi-voltage stimulation of the ACC (n = 7) and paralimbic subcaudate (n = 5) regions were recorded during bilateral anterior cingulotomy and bilateral subcaudate tractotomy (in patients that had previously received an adequate lesion in the ACC), respectively. RESULTS: Stimulations in both groups were accompanied by increased autonomic arousal. Skin conductance activity was significantly increased during ACC stimulations compared with paralimbic targets at 2 V (2.34 +/- .68 [score in microSiemens +/- SE] vs. .34 +/- .09, p = .013) and 3 V (3.52 +/- .86 vs. 1.12 +/- .37, p = .036), exhibiting a strong "voltage response" relationship between stimulus magnitude and response amplitude (difference from 1 to 3 V = 1.15 +/- .90 vs. 3.52 +/- .86, p = .041). Heart rate response was less indicative of between-group differences. CONCLUSIONS: This is the first study of its kind aiming at seeking novel insights into the mechanisms responsible for central autonomic modulation. It supports a concept that interregional interactions account for the coordination of autonomic arousal. PMID- 19545861 TI - Synthesis and characterization of a crosslinked chitosan derivative with a complexing agent and its adsorption studies toward metal(II) ions. AB - A new chitosan derivative has been synthesized by crosslinking a metal complexing agent, [6,6'-piperazine-1,4-diyldimethylenebis (4-methyl-2-formyl) phenol] (L), with chitosan (CTS). The resulting material (CCTSL) was characterized by elemental (CHN), spectral (FTIR and solid-state NMR), thermal (TGA and DTA), and structural (powder XRD and SEM) analyses. Adsorption experiments (pH dependency, kinetics, and equilibrium) of CCTSL toward various metal ions such as Mn(II), Fe(II), Co(II), Cu(II), Ni(II), Cd(II), and Pb(II) were carried out at 25 degrees C. The results showed that the adsorption was dependent on the pH of the solution, with a maximum capacity between pHs 6.5 and 8.5. The kinetics was evaluated by applying the pseudo-first-order and pseudo-second-order equation models and the equilibrium data were analyzed by Langmuir isotherm model. The maximum adsorption capacity was 1.21 mmol g(-1) for Cu(II) and the order of adsorption capacities for the metal(II) ions studied was found to be Cu(II)>Ni(II)>Cd(II)> or =Co(II)> or =Mn(II)> or =Fe(II)> or =Pb(II). PMID- 19545860 TI - Autism and nonsyndromic mental retardation associated with a de novo mutation in the NLGN4X gene promoter causing an increased expression level. AB - BACKGROUND: Pathogenic mutations in the X-linked Neuroligin 4 gene (NLGN4X) in autism spectrum disorders (ASDs) and/or mental retardation (MR) are rare. However, nothing is known regarding a possible altered expression level of NLGN4X that would be caused by mutations in regulatory sequences. We investigated this issue by analyzing these regions in patients with ASDs and no mutation in the NLGN4X coding sequence. METHODS: We studied 96 patients who met all DSM-IV criteria for autism. The entire coding sequence and the regulatory sequences of the NLGN4X gene were analyzed by polymerase chain reaction and direct sequencing. RESULTS: We identified a de novo 1 base pair (-335G>A) substitution located in the promoter region in a patient with autism and nonsyndromic profound MR. Interestingly, this variation is associated with an increased level of the NLGN4X transcript in the patient compared with male control subjects as well as his father. Further in vitro luciferase reporter and electrophoretic mobility shift assays confirmed, respectively, that this mutation increases gene expression and is probably caused by altered binding of transcription factors in the mutated promoter sequence. CONCLUSIONS: This result brings further insight about the phenotypic spectrum of NLGN4X mutations and suggests that the analysis of the expression level of NLGN4X might detect new cases. PMID- 19545863 TI - [Controversies in living donor liver transplants]. AB - Adult liver transplants from a living donor are a valid and effective alternative as a treatment in terminal liver disease. However, in the context of a donation from a brain dead donor, as in western countries (Spain), it is very high, and there are still controversial aspects that should be continuously assessed in order to ensure good results. Live donations are currently stable and represent about 5% of the total liver transplants performed in Europe. Morbidity and mortality is around 35% and 0.1%, respectively, probably reasonable figures given the characteristics of the intervention. The 1 and 5 year survival rates of the recipients of a living donor are currently 95% and 75%, which are similar to those coming from brain dead donors. However the level of biliary complications in this patient group is higher, with an incidence of around 35-40%. However, this incidence has not had any effect on the long-term results up until now. PMID- 19545862 TI - Comparative analysis of DC fused with allogeneic hepatocellular carcinoma cell line HepG2 and autologous tumor cells as potential cancer vaccines against hepatocellular carcinoma. AB - Fusions of patient-derived dendritic cells (DCs) and autologous tumor cells induce T-cell responses against autologous tumors in animal models and human clinical trials. These fusion cells require patient-derived tumor cells, which are not, however, always available. Here we fused autologous DCs from patients with hepatocellular carcinoma (HCC) to an allogeneic HCC cell line (HepG2). These fusion cells co-expressed tumor-associated antigens (TAAs) and DC-derived costimulatory and MHC molecules. Both CD4(+) and CD8(+) T cells were activated by the fusion cells. Cytotoxic T lymphocytes (CTLs) induced by the fusion cells were able to kill autologous HCC by HLA-A2- and/or HLA-A24-restricted mechanisms. CTL activity against shared TAAs indicates that the presence of alloantigens does not prevent the development of CTLs with activity against autologous HCC cells. These fusion cells may have applications in anti-tumor immunotherapy through cross priming against shared tumor antigens and may provide a platform for adoptive immunotherapy. PMID- 19545864 TI - The metamorphosis of the statistical segmentation output: lexicalization during artificial language learning. AB - This study combined artificial language learning (ALL) with conventional experimental techniques to test whether statistical speech segmentation outputs are integrated into adult listeners' mental lexicon. Lexicalization was assessed through inhibitory effects of novel neighbors (created by the parsing process) on auditory lexical decisions to real words. Both immediately after familiarization and post-one week, ALL outputs were lexicalized only when the cues available during familiarization (transitional probabilities and wordlikeness) suggested the same parsing (Experiments 1 and 3). No lexicalization effect occurred with incongruent cues (Experiments 2 and 4). Yet, ALL differed from chance, suggesting a dissociation between item knowledge and lexicalization. Similarly contrasted results were found when frequency of occurrence of the stimuli was equated during familiarization (Experiments 3 and 4). Our findings thus indicate that ALL outputs may be lexicalized as far as the segmentation cues are congruent, and that this process cannot be accounted for by raw frequency. PMID- 19545866 TI - The evolution of Mexico City's abortion laws: from public morality to women's autonomy. AB - Before 2000, Mexico City's criminal laws prohibited induced abortion to maintain public morality. The Criminal Code considered abortion by accident or in cases of rape not criminal, and criminal but excusable-and therefore not punishable-in certain cases not endangering public morality, such as medical necessity to save the woman's life. In 2000, the Criminal Code was reformed expanding exceptions from criminal liability, particularly in cases of danger to a woman's health or where fetal survival was at risk. In 2004, Mexico City enacted its own law, effectively decriminalizing consensual abortion in cases of rape, fetal malformation, and risk to the woman's health. A 2007 reform further decriminalized all consensual abortion within the first 12 weeks of pregnancy, and required public hospitals to provide abortion and family planning services. In August 2008, the Supreme Court of Mexico ruled Mexico City's 2007 liberalization of abortion law constitutional. PMID- 19545865 TI - Validity of a traffic air pollutant dispersion model to assess exposure to fine particles. AB - INTRODUCTION: Fine particles (PM(2.5)) are an important component of air pollution. Epidemiological studies have shown health effects due to ambient air particles, particularly allergies in children. Since the main difficulty is to determine exposure to such pollution, traffic air pollutant (TAP) dispersions models have been developed to improve the estimation of individual exposure levels. One such model, the ExTra index, has been validated for nitrogen oxide concentrations but not for other pollutants. The purpose of this study was to assess the validity of the ExTra index to assess PM(2.5) exposure. METHODS: We compared PM(2.5) concentrations calculated by the ExTra index to reference measures (passive samplers situated under the covered part of the playground), in 15 schools in Bordeaux, in 2000. First, we collected the input data required by the ExTra index: background and local pollution depending on traffic, meteorology and topography. Second, the ExTra index was calculated for each school. Statistical analysis consisted of a graphic description; then, we calculated an intraclass correlation coefficient. RESULTS: Concentrations calculated with the ExTra index and the reference method were similar. The ExTra index underestimated exposure by 2.2 microg m(-3) on average compared to the reference method. The intraclass correlation coefficient was 0.85 and its 95% confidence interval was [0.62; 0.95]. CONCLUSIONS: The results suggest that the ExTra index provides an assessment of PM(2.5) exposure similar to that of the reference method. Although caution is required in interpreting these results owing to the small number of sites, the ExTra index could be a useful epidemiological tool for reconstructing individual exposure, an important challenge in epidemiology. PMID- 19545867 TI - Papillary serous carcinoma in situ in ovarian endometriosis in an MSH2 mutation carrier. PMID- 19545868 TI - [Evidence-based medicine or factual medicine]. PMID- 19545869 TI - Association of moderate and severe hypertriglyceridemia with obesity, diabetes mellitus and vascular disease in the Spanish working population: results of the ICARIA study. AB - AIM: To study the prevalence, risk factors, and vascular disease associated with moderate and severe hypertriglyceridemia in an active working population. DESIGN AND METHODS: Cross-sectional study of 594,701 workers from all Spanish geographical areas, occupation sectors, ages, and sexes who underwent a yearly routine checkup. Data collected from participants included age, sex, anthropometric measurements, vascular risk factors, lipidic profile and basic biochemical analysis, from a fasting blood sample. A cardiovascular risk assessment was performed. RESULTS: The study population included 428,334 males and 166,367 females, mean age 36+/-10 years. A total of 95,673 (16%) workers had mild hypertriglyceridemia (HTg) (Tg 150-399mg/dL), 7,081 (1.1%) had moderate HTg (400-999mg/dL), and 224 (0.03%) had severe HTg (>or=1000mg/dL). Of workers with hypertriglyceridemia, 90% were male. Age, obesity, type 1 and 2 diabetes, alcohol consumption, and vascular disease were associated with hypertriglyceridemia. Cardiovascular risk gradually increased for each HTg category. Amongst risk factors, the major independent predictor of mild-HTg was obesity (OR 2.42, CI 95% 2.37-2.48), whereas diabetes was a predictor of moderate HTg (OR 3.64, CI 95% 3.17-4.18) and severe HTg (OR 7.35, CI 95% 4.27-12.66). In multivariate analyses, HTg was gradually associated with vascular disease, even after adjusting for other risk factors. CONCLUSION: In this working population, preventive programs for HTg and associated vascular disease should consider obesity-diabetes control as its first objective. PMID- 19545871 TI - Tissue factor genetic polymorphisms and haplotypes in Japanese population. PMID- 19545870 TI - Management of non-high-density lipoprotein abnormalities. AB - Epidemiological evidence supports the use of non-high-density lipoprotein cholesterol (non-HDL-C), apolipoprotein B-100 (apoB), and low-density lipoprotein particles as markers of atherogenic risk. Treatment guidelines also identify these as additional targets of lipid-modifying intervention in patients with elevated triglycerides (TG). Even when TG are only moderately elevated, many patients on statin monotherapy who have achieved targets for low-density lipoprotein cholesterol (LDL-C) fail to reach non-HDL-C treatment goals, and even fewer reach apoB goals. Combination lipid-modifying therapy is therefore indicated for comprehensive lipid management, particularly in patients with type 2 diabetes and metabolic syndrome in whom LDL-C levels are often considered 'optimal'. Of the available options, adding either a niacin, fibrate or omega-3 fatty acids provides greater opportunity to achieve non-HDL-C and apoB targets, given complementary profiles of lipid-modifying activity and supported by evidence from clinical studies. Improvement in lipid control and reduction in atherogenic risk could be anticipated to translate to benefits in clinical outcomes. PMID- 19545872 TI - Preparative size exclusion chromatography combined with detergent removal as a versatile tool to prepare unilamellar and spherical liposomes of highly uniform size distribution. AB - Detergent removal from mixed micelles was combined with preparative size exclusion chromatography (SEC) on Sephacryl S 500 HR to prepare unilamellar and spherical liposomes of defined sizes between 50 and 100 nm with a very narrow size distribution (RSD of vesicle diameter between 13% and 25%). For neutral phosphatidylcholine and negatively charged phosphatidylcholine/phosphatidylglycerol liposome preparations, efficient sizing at the preparative scale was demonstrated by analyzing isolated SEC peak fractions with cryo-transmission electron microscopy and dynamic light scattering. The number-weighted average vesicle diameters obtained using both methods are in very good agreement for fractions of low polydispersity. PMID- 19545873 TI - Particle concentration measurement of virus samples using electrospray differential mobility analysis and quantitative amino acid analysis. AB - Virus reference materials are needed to develop and calibrate detection devices and instruments. We used electrospray differential mobility analysis (ES-DMA) and quantitative amino acid analysis (AAA) to determine the particle concentration of three small model viruses (bacteriophages MS2, PP7, and phiX174). The biological activity, purity, and aggregation of the virus samples were measured using plaque assays, denaturing gel electrophoresis, and size-exclusion chromatography. ES-DMA was developed to count the virus particles using gold nanoparticles as internal standards. ES-DMA additionally provides quantitative measurement of the size and extent of aggregation in the virus samples. Quantitative AAA was also used to determine the mass of the viral proteins in the pure virus samples. The samples were hydrolyzed and the masses of the well-recovered amino acids were used to calculate the equivalent concentration of viral particles in the samples. The concentration of the virus samples determined by ES-DMA was in good agreement with the concentration predicted by AAA for these purified samples. The advantages and limitations of ES-DMA and AAA to characterize virus reference materials are discussed. PMID- 19545874 TI - Fast separation of selective serotonin reuptake inhibitors antidepressants in plasma sample by nonaqueous capillary electrophoresis. AB - A simple, fast, and sensitive liquid-liquid extraction method followed by nonaqueous capillary electrophoresis (LLE/NACE) was developed and validated for simultaneous determination of four antidepressants (fluoxetine, sertraline, citalopram and paroxetine) in human plasma. Several experimental separation conditions using aqueous and nonaqueous media separation were tested by varying the electrolyte pH value (for aqueous medium) and the ionic strength concentration considering the similar mobility of the compounds. High-resolution separation was achieved with a mixture of 1.25 mol L(-1) of phosphoric acid in acetonitrile. The quantification limits of the LLE/CE method varied between 15 and 30 ng mL(-1), with a relative standard deviation (RSD) lower than 10.3%. The method was successfully applied in therapeutic drug monitoring and should be employed in the evaluation of plasma levels in urgent toxicological analysis. PMID- 19545875 TI - Determination of nerve agent degradation products by capillary electrophoresis using field-amplified sample stacking injection with the electroosmotic flow pump and contactless conductivity detection. AB - In the present study, field-amplified sample stacking injection using the electroosmotic flow pump (FAEP) was developed for the capillary electrophoretic separation of the four nerve agent degradation products methylphosphonic acid (MPA), ethyl methylphosphonic acid (EMPA), isopropyl methylphosphonic acid (IMPA) and cyclohexyl methylphosphonic acid (CMPA). Coupled to contactless conductivity detection, direct quantification of these non-UV active compounds could be achieved. Sensitivity enhancement of up to 500 to 750-fold could be obtained. The newly established approach was applied to the determination of the analytes in river water and aqueous extracts of soil. Detection limits of 0.5, 0.7, 1.4 and 2.7 ng/mL were obtained for MPA, EMPA, IMPA and CMPA, respectively, in river water and 0.09, 0.14, 0.44 and 0.22 microg/g, respectively, in soil. PMID- 19545876 TI - Effect of the direction of ester linkage on molecular shape selectivity through multiple carbonyl-pi interaction with octadecyl chain branched polymers as organic phases in reversed-phase high-performance liquid chromatography. AB - Poly(vinyl octadecanoate)-grafted porous silica (Sil-VODn, n=23) was newly prepared to investigate the efficiencies of the carbonyl groups in the polymer chain for recognition of polycyclic aromatic hydrocarbons (PAHs) in RP-HPLC. In Sil-VOD23, the octadecyl side chains were connected to the polymer main chain through ester linkage in opposite direction to that in poly(octadecylacrylate) grafted silica (Sil-ODAn, n=25) which has been reported by us. Sil-ODAn performs enhanced molecular shape selectivity of PAHs in RP-HPLC through multiple carbonyl pi interaction of aligned carbonyl groups which are induced by the formation of highly oriented structure of side chains. Differential scanning calorimetry of VOD23 demonstrated that octadecyl alkyl chains showed crystalline to isotropic phase transition with endothermic peak at 48.7 degrees C which was similar to ODA25 (at 47.8 degrees C). After grafting of both polymers, phase transition phenomenon was completely disappeared in Sil-VOD23 whereas Sil-ODA25 still exhibits phase transition although at lower endothermic peak top temperature (38.5 degrees C). This indicates that the slight structural change in Sil-VODn and Sil-ODAn influence the ordered structure of side alkyl chains. Moreover, solid-state 13C NMR revealed that the long alkyl chain in Sil-VOD23 is highly disordered as compared with that of Sil-ODA25. Sil-VOD23 was applied to RP-HPLC stationary phase using PAHs as pi-electron containing elutes, and compared with Sil-ODA25 and conventional monomeric octadecylated silica (ODS). Results confirmed that Sil-VOD23 showed much higher selectivity for PAH isomers than ODS, but lower than Sil-ODA25. For example, the separation factors for trans-/cis stilbene were 1.47 (Sil-VOD23), 1.70 (Sil-ODA25) and 1.07 (ODS), respectively. These results indicate that carbonyl groups in Sil-VOD23 are effective for molecular shape recognition of PAHs through carbonyl-pi interactions even in the disordered state. PMID- 19545877 TI - Thermoresponsive surfaces prepared using adsorption of a cationic graft copolymer: a versatile method for triggered particle capture. AB - In this study we investigate triggered particle capture at substrates containing adsorbed thermally responsive graft copolymers. The copolymers used were PDMA(x)(+)-g-(PNIPAm(n))(y), where DMA(+) is quaternized N,N-dimethylaminoethyl methacrylate and NIPAm is N-isopropylacrylamide. The x and y values originate from the macroinitiator used for copolymer preparation. In this study the copolymers are adsorbed onto two different substrates: quartz microscope slides and microporous, high surface area carbon foam. The substrates were coated with a layer of calcined laponite. The laponite acted as a conditioning layer and promoted strong adsorption of the copolymer. The hydrophobicity of the thermoresponsive surfaces was probed using variable-temperature contact angle measurements. The contact angles generally increased considerably upon increasing the temperature to above the lower critical solution temperature (LCST) of the copolymers. The ability of the thermoresponsive surfaces to capture dispersed particles was investigated using anionic and cationic polystyrene (PS) particles. PDMA(30)(+)-g-(PNIPAm(210))(14) was the most effective copolymer in terms of providing high capture efficiencies of anionic PS particles using temperature as the trigger. The thermoresponsive surfaces strongly held the anionic PS particles even when cooled to below the LCST. The relationships between copolymer structure and particle capture efficiency are discussed. The new approach used here for preparation thermoresponsive surfaces is potentially scalable to high volume applications. PMID- 19545878 TI - V2O5 nanoparticles obtained from a synthetic bariandite-like vanadium oxide: synthesis, characterization and electrochemical behavior in an ionic liquid. AB - Highly stable and crystalline V(2)O(5) nanoparticles with an average diameter of 15 nm have been easily prepared by thermal treatment of a bariandite-like vanadium oxide, V(10)O(24) x 9 H(2)O. Their characterization was carried out by powder X-ray diffractometry (XRD), Fourier transform infrared (FT-IR) and Raman spectroscopies, and transmission electron microscopy (TEM). The fibrous and nanostructured film obtained by electrophoretic deposition of the V(2)O(5) nanoparticles showed good electroactivity when submitted to cyclic voltammetry in an ionic liquid-based electrolyte. The use of this film for the preparation of a nanostructured electrode led to an improvement of about 50% in discharge capacity values when compared with similar electrodes obtained by casting of a V(2)O(5) xerogel. PMID- 19545879 TI - Specific thermodynamical potentials on surfaces under relativistic conditions. AB - Expressions of specific thermodynamical potentials on surfaces (J/cm(2)) were obtained (for internal energy, enthalpy, free energy, and free enthalpy) under relativistic conditions. PMID- 19545880 TI - Electrocatalytic properties of graphite nanofibers-supported platinum catalysts for direct methanol fuel cells. AB - Graphite nanofibers (GNFs) treated at various temperatures were used as carbon supports to improve the efficiency of PtRu catalysts. The electrochemical properties of the PtRu/GNFs catalysts were then investigated to evaluate their potential for application in DMFCs. The results indicated that the particle size and dispersibility of PtRu in the catalysts were changed by heat treatment, and the electrochemical activity of the catalysts was improved. Consequently, it was found that heat treatments could have an influence on the surface and structural properties of GNFs, resulting in enhancing an electrocatalytic activity of the catalysts for DMFCs. PMID- 19545881 TI - Pyrrolizidine alkaloid biosynthesis, evolution of a pathway in plant secondary metabolism. AB - The system of pyrrolizidine alkaloids has proven to be a powerful system for studying the evolution of a biosynthetic pathway in plant secondary metabolism. Pyrrolizidine alkaloids are typical plant secondary products produced by the plant as a defense against herbivores. The first specific enzyme, homospermidine synthase, has been shown to have evolved by duplication of the gene encoding deoxyhypusine synthase, which is involved in primary metabolism. Despite the identical function of homospermidine synthase for pyrrolizidine alkaloid biosynthesis in the various plant lineages, this gene duplication has occurred several times independently during angiosperm evolution. After duplication, these gene copies diverged with respect to gene function and regulation. In the diverse plant lineages producing pyrrolizidine alkaloids, homospermidine synthase has been shown to be expressed in a variety of tissues, suggesting that the regulatory elements were recruited individually after the duplication of the structural gene. The molecular, kinetic, and expression data of this system are discussed with respect to current models of gene and pathway evolution. PMID- 19545882 TI - The prevalence and distribution of Mycobacterium bovis infection in European badgers (Meles meles) as determined by enhanced post mortem examination and bacteriological culture. AB - The accurate diagnosis of Mycobacterium bovis infection in badgers is key to understanding the epidemiology of tuberculosis in this species and has significant implications for devising strategies to limit spread of the disease. In this study, badgers (n=215) in the Republic of Ireland were examined at post mortem and tissues were collected from a range of anatomical locations and pooled into groups for bacterial culture of M. bovis. By assessing confirmed gross visible lesions (VL) alone, infection was detected in 12.1% of badgers. However, by including the results of all culture positive pooled samples, the overall infection prevalence increased significantly to 36.3%. Two-thirds (66.7%) of infected animals had no visible lesions (NVL). While the thoracic cavity (lungs and pulmonary lymph nodes) was found to be the most common site of infection, in a proportion of animals infection was absent from the lungs and draining lymph nodes and was confined to the lymph nodes of the carcase or the head. This may indicate an early extrapulmonary dissemination of infection or alternatively, in the case of the head lymph nodes, a secondary pathogenic pathway involving the lymphoid tissues of the upper respiratory tract (URT). PMID- 19545883 TI - Analysis of coherent surface wave dispersion and attenuation for non-destructive testing of concrete. AB - Rayleigh waves measurements are used to characterise cover concrete and mortar in the frequency range 60-180 kHz. At these frequencies, the wavelength is comparable to the size of the aggregates, and waves propagate in a multiple scattering regime. Acquired signals are then difficult to interpret due to an important incoherent part. The method proposed here is the study of the coherent waves, obtained by averaging signals over several configurations of disorder. Coherent waves give information on an equivalent homogeneous medium. To acquire a large amount of measurements with accuracy, an optimised piezoelectric source is used with a laser interferometer for reception. Adapted signal processing technique are presented to evaluate the coherent phase and group velocities and also the coherent attenuation parameter. The sensitivity of these three parameters with the properties of concrete is discussed, as well as the necessity to use coherent waves to obtain accurate results. PMID- 19545884 TI - Towards classification of the bifurcation structure of a spherical cavitation bubble. AB - We focus on a single cavitation bubble driven by ultrasound, a system which is a specimen of forced nonlinear oscillators and is characterized by its extreme sensitivity to the initial conditions. The driven radial oscillations of the bubble are considered to be implicated by the principles of chaos physics and owing to specific ranges of control parameters, can be periodic or chaotic. Despite the growing number of investigations on its dynamics, there is not yet an inclusive yardstick to sort the dynamical behavior of the bubble into classes; also, the response oscillations are so complex that long term prediction on the behavior becomes difficult to accomplish. In this study, the nonlinear dynamics of a bubble oscillator was treated numerically and the simulations were proceeded with bifurcation diagrams. The calculated bifurcation diagrams were compared in an attempt to classify the bubble dynamic characteristics when varying the control parameters. The comparison reveals distinctive bifurcation patterns as a consequence of driving the systems with unequal ratios of R(0)lambda (where R(0) is the bubble initial radius and lambda is the wavelength of the driving ultrasonic wave). Results indicated that systems having the equal ratio of R(0)lambda, share remarkable similarities in their bifurcating behavior and can be classified under a unit category. PMID- 19545885 TI - Ozonation of pharmaceutical compounds: Rate constants and elimination in various water matrices. AB - The ozonation of four pharmaceuticals (metoprolol, naproxen, amoxicillin, and phenacetin) in ultra-pure (UP) water was studied in the pH range between 2.5 and 9. The experiments allowed the determination of the apparent rate constants for the reactions between ozone and the selected compounds. The values obtained varied depending on the pH, and ranged between 239 and 1.27x10(4)M(-1) s(-1) for metoprolol; 2.62x10(4) and 2.97x10(5)M(-1)s(-1) for naproxen; 2.31x10(3) and 1.21x10(7)M(-1)s(-1) for amoxicillin; and 215 and 1.57x10(3)M(-1)s(-1) for phenacetin. Due to the acidic nature of these substances, the degree of dissociation of each pharmaceutical was determined at every pH of work, and the specific rate constants of the neutral and ionic species formed were evaluated. Additionally, the simultaneous ozonation of the pharmaceuticals in different water matrices was carried out by considering a groundwater, a surface water from a public reservoir, and three secondary effluents from municipal wastewater treatment plants. The influence of the operating conditions (initial ozone dose, nature of pharmaceuticals and type of water) on the pharmaceuticals elimination efficiency was established, and a kinetic model was proposed for the evaluation of the partial contribution to the global oxidation of both, the direct ozonation reaction and the radical pathway. PMID- 19545886 TI - Reduction of natural organic matter by nanofiltration process. AB - The objectives of this investigation were to characterize natural organic matter (NOM) in Kin-men raw water, to evaluate the performance of the nanofiltration (NF) membrane process for NOM removal, and to determine the good engineering practice (GEP) of the NF 270 membrane filtration performance. Three different samples: (1) raw water collected from the Tai Lake in Kin-men, Taiwan; (2) ultrafiltration (UF) pre-treated water; and (3) sand filtration (SF) treated water after the coagulation and sedimentation processes were employed in this study. The hydrophobic component (58%) was the predominant NOM fraction, and the NOM molecular weight was distributed broadly, i.e., lower than 1 kDa (30%), 1-5 kDa (32%) and larger than 5 kDa (38%). Hydrophobic NOM easily accumulated on the NF membrane surface and resulted in flux decline. The SF-NF was the proposed treatment process because it can reduce the NOMs effectively with lower energy consumption than UF-NF. It was noted that the dissolved organic carbon (DOC) rejection ratio was not affected by changing cross-flow velocity. However, at lower cross-flow velocity (0.15 ms(-1)), the reduction of organic matter in water sample was higher than that at higher cross-flow velocity (0.30 ms(-1)). By integrating the experimental results, it was concluded that a transmembrane pressure of 690 kPa and a cross-flow velocity of 0.30 ms(-1) exhibited the GEP, yielding about 94% of both DOC and UV(254) reduction. PMID- 19545887 TI - Phthalate acid esters in Potamogeton crispus L. from Haihe River, China. AB - This study was conducted in Haihe River, China, in order to assess the ability of Potamogeton crispus L. (P. crispus), a submerged plant, to accumulate phthalic acid esters (PAEs). Dibutyl phthalate (DBP) and di-2-ethylexyl phthalate (DEHP) were measured in samples of water, sediment and P. crispus plant from March to May 2008. The results showed that the highest levels of DBP and DEHP in above ground tissues appeared during the blooming period of P. crispus, which was consistent with the results obtained with the water samples. Regression analysis reveals that concentrations of PAEs in above-ground tissues were mainly influenced by PAE concentrations in the water of Haihe River. Enrichment of DBP and DEHP in above-ground tissues was observed, with bioconcentration factors (BCFs) of 4.82-83.65Lkg(-1) for DBP and 6.71-93.70Lkg(-1) for DEHP. The distribution pattern of DBP in roots, surface and near root sediments was different from that of DEHP. Concentrations of DBP in near root sediments were lower than those in roots and comparable to those in surface sediments, while concentrations of DEHP in roots were lower than those in near root sediments but higher than those in surface sediments. PMID- 19545888 TI - Preparation of chitosan-stabilized Fe(0) nanoparticles for removal of hexavalent chromium in water. AB - Chitosan-stabilized Fe(0) nanoparticles (CTO-Fe(0)) and Fe(0) nanoparticles synthesized in ethanol-water mixed system (EW-Fe(0)) were tested for reduction of Cr(VI) in water. Fourier transform infrared (FTIR) study suggested that nitrogen and oxygen atoms are the binding sites for chitosan on iron which was accountable for the stability of Fe(0) nanoparticles. While the EW-Fe(0) ignites spontaneously when exposed to air, the CTO-Fe(0) was still in zero valence state after exposure to air over 2-month period as shown by X-ray powder diffraction patterns. Batch experiments demonstrated that the maximum Cr(VI) reduction rates for CTO-Fe(0) was about 3 times higher than EW-Fe(0). Characterizations with high resolution X-ray photoelectron spectroscopy (HR-XPS) revealed that Cr(VI) was reduced to Cr(III) and Fe(III) was the only component present on the Fe(0) nanoparticles surface. Additionally, chitosan can inhibited the formation of Fe(III)-Cr(III) precipitation due to its high ability to chelate Fe(III) which resulted in k(obs) for CTO-Fe(0) was about 1-3 times higher than EW-Fe(0). Due to the fast reaction kinetics and good stability against oxidation in air, the chitosan-stabilized Fe(0) nanoparticles have the potential to become an effective agent for in situ subsurface environment remediation. PMID- 19545889 TI - In vitro and in vivo evaluation of akermanite bioceramics for bone regeneration. AB - This study investigated the effects of a calcium magnesium silicate bioceramic (akermanite) for bone regeneration in vitro and in vivo, with beta-tricalcium phosphate (beta-TCP) as a control. In vitro, the human bone marrow-derived mesenchymal stromal cells (hBMSCs) were cultured in an osteogenic medium supplemented with a certain concentration of two bioceramics' extracts for 20 days. An MTT assay showed that akermanite extract promoted proliferation of hBMSC significantly more than did beta-TCP extract. The results of alkaline phosphatase (ALP) activity test and the expression of osteogenic marker genes such as ALP, osteopontin (OPN), osteocalcin (OCN) and bone sialoprotein (BSP) demonstrated that the osteogenic differentiation of hBMSC was enhanced more by akermanite extract than by beta-TCP extract. In vivo, a histomorphology analysis and histomorphometry of the two porous bioceramics implants in rabbit femur defect models indicated that both in early- and late-stage implantations, akermanite promoted more osteogenesis and biodegradation than did beta-TCP; and in late stage implantations, the rate of new bone formation was faster in akermanite than in beta-TCP. These results suggest that akermanite might be a potential and attractive bioceramic for tissue engineering. PMID- 19545890 TI - A paradigm for peptide vaccine delivery using viral epitopes encapsulated in degradable polymer hydrogel capsules. AB - We report on the use of degradable polymer capsules as carriers for the delivery of oligopeptide antigens to professional antigen presenting cells (APCs). To achieve encapsulation, oligopeptide sequences were covalently linked to a negatively charged carrier polymer via biodegradable linkages and the resulting conjugate was then adsorbed onto amine-functionalized silica particles. These peptide-coated particles were then used as templates for the layer-by-layer (LbL) deposition of thiolated poly(methacrylic acid) (PMA(SH)) and poly(vinylpyrrolidone) (PVPON) multilayers. Removal of the silica core and disruption of the hydrogen bonding between PMA(SH) and PVPON by altering the solution pH yielded disulfide-stabilized PMA capsules that retain the encapsulated cargo in an oxidative environment. In the presence of a natural reducing agent, glutathione, cleavage of the disulfide bonds causes release of the peptide from the capsules. The developed strategy provides control over peptide loading into polymer capsules and yields colloidally stable micron- and submicron-sized carriers with uniform size and peptide loading. The conjugation and encapsulation procedures were proven to be non-degrading to the peptide vaccines. The peptide-loaded capsules were successfully used to deliver their cargo to APCs and activate CD8 T lymphocytes in a non-human primate model of SIV infection ex vivo. The reported approach represents a novel paradigm in the delivery of peptide vaccines and other therapeutic agents. PMID- 19545891 TI - Integrating polyurethane culture substrates into poly(dimethylsiloxane) microdevices. AB - Poly(dimethylsiloxane) (PDMS)-based microdevices have enabled rapid, high throughput assessment of cellular response to precisely controlled microenvironmental stimuli, including chemical, matrix and mechanical factors. However, the use of PDMS as a culture substrate precludes long-term culture and may significantly impact cell response. Here we describe a method to integrate polyurethane (PU), a well-studied and clinically relevant biomaterial, into the PDMS multilayer microfabrication process, enabling the exploration of long-term cellular response on alternative substrates in microdevices. To demonstrate the utility of these hybrid microdevices for cell culture, we compared initial cell adhesion, cell spreading, and maintenance of protein patterns on PU and PDMS substrates. Initial cell adhesion and cell spreading after three days were comparable between collagen-coated PDMS and PU substrates (with or without collagen coating), but significantly lower on native PDMS substrates. However, for longer culture durations (> or = 6 days), cell spreading and protein adhesion on PU substrates was significantly better than that on PDMS substrates, and comparable to that on tissue culture-treated polystyrene. Thus, the use of a generic polyurethane substrate in microdevices enables longer-term cell culture than is possible with PDMS substrates. More generally, this technique can improve the impact and applicability of microdevice-based research by facilitating the use of alternate, relevant biomaterials while maintaining the advantages of using PDMS for microdevice fabrication. PMID- 19545892 TI - Conformational recovery and preservation of protein nature from heat-induced denaturation by water-soluble phospholipid polymer conjugation. AB - The effect of water-soluble phospholipid polymer conjugated to a protein on conformational change during heat-quenched stress was investigated in this study. Well-defined pyridine disulfide end-functional poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) was synthesized by well controlled atom transfer radical polymerization method. Synthesized PMPC was the site specifically conjugated to a protein, and the effect on conformational change during heat quenched stress was estimated by circular dichroism and fluorescence study. As a result, a single phospholipid polymer chain conjugated to a protein give a great effect on conformational preservation in both secondary and tertiary structures even after the heat-quenched process. Moreover, even the conformational recovery which gave the completely reversible conformational preservation was observed in circular dichroism study. The resulting protein activity was also confirmed, and no significant decline was induced by heat-quenched stress. PMID- 19545893 TI - Influences of protein films on antibacterial or bacteria-repellent surface coatings in a model system using silicon wafers. AB - Immobilization of defined chemical functionalities to biomaterial surfaces is employed to optimize them not only for tissue compatibility but also for prevention of bacterial infection. Grafting surfaces with chains of poly(ethylene glycol) (PEG) results in bacterial repellence whereas modification with cationic groups conveys them with bactericidal properties. Since biomaterials in situ will become exposed to a protein-rich environment, it is necessary to investigate the influence of prior protein adsorption on the antibacterial activity of this type of chemical surface modification. In the present study, we immobilized short chain PEG and two pyridinium group-containing methacrylate monomers, 12 methacryloyloxydodecylpyridinium bromide (MDPB) and 6 methacryloyloxyhexylpyridinium chloride (MHPC), to silicon wafer model surfaces to investigate the influence of prior protein adsorption on the bactericidal activity of the surface coating towards subsequently attached bacteria. Adsorbed amounts of human serum albumin and salivary proteins were found to be two times higher on cationic compared to PEG-modified surfaces. An analogous tendency was found for attachment of Streptococcus gordonii and Streptococcus mutans to the same surfaces without prior protein exposure. However, most bacteria attached to cationic surfaces were found to be dead. Prior exposure of cationic surfaces to protein solutions drastically altered bacterial attachment dependent on the type of protein solution and bacterial species employed. Significantly, the original bactericidal activity of pyridinium-coated surfaces was found greatly reduced upon adsorption of a protein film. As a conclusion we propose that future approaches should combine the protein- and bacteria-repellent properties of PEG coatings with the bactericidal function of charged cationic groups. PMID- 19545894 TI - Nanoscale presentation of cell adhesive molecules via block copolymer self assembly. AB - Precise control over the nanoscale presentation of adhesion molecules and other biological factors represents a new frontier for biomaterials science. Recently, the control of integrin spacing and cellular shape has been shown to affect fundamental biological processes, such as differentiation and apoptosis. Here, we present the self-assembly of maleimide functionalised polystyrene-block-poly (ethylene oxide) copolymers as a simple, yet highly precise method for controlling the position of cellular adhesion molecules. By manipulating the phase separation of the functional PS-PEO block copolymer used in this study, via a simple blending technique, we alter the nanoscale (on PEO domains of 8-14 nm in size) presentation of the adhesion peptide, GRGDS, decreasing lateral spacing from 62 nm to 44 nm and increasing the number density from approximately 450 to approximately 900 islands per microm2. The results indicate that the spreading of NIH-3T3 fibroblasts increases as the spacing between domains of RGD binding peptides decreases. Further, the same functional PS-PEO surfaces have been utilised to immobilise, via a zinc chelating peptide sequence, poly-histidine tagged proteins and extracellular matrix (ECM) fragments. This method is seen as an ideal platform for investigations into the role of spatial arrangements of cell adhesion molecules and ECM molecules on cell function and, in particular, control of cell phenotype. PMID- 19545895 TI - Placental dysferlin expression is reduced in severe preeclampsia. AB - Dysferlin (DYSF) and myoferlin (MYOF), members of the ferlin family of membrane proteins, are co-expressed in human placental syncytiotrophoblast (STB). Although the role of these ferlin proteins in the placenta has yet to be established, it has been suggested that DYSF and MYOF may contribute to the stability of the apical STB plasma membrane. The release of STB-derived cellular debris increases in the setting of preeclampsia (PE), suggesting relative destabilization of the hemochorial interface. To test whether PE was associated with alterations in placental expression of DYSF and/or MYOF, a cross-sectional study was performed using specimens of villous placenta collected form women with severe PE (n=10) and normotensive controls (n=10). DYSF and MYOF expression were examined using quantitative real-time RT-PCR, immunoblotting, and immunofluorescence labeling of tissue specimens. Placental DYSF expression was 57% lower at the mRNA level (p=0.03) and 38% lower at the protein level (p=0.026) in severe PE as compared to normotensive subjects. There were no differences in placental MYOF protein or mRNA expression between these groups. No appreciable changes in the distribution of DYSF or MYOF within placental villi was observed in PE relative to control specimens. We conclude that DYSF expression is reduced in severe PE relative to gestational age-matched controls. As DYSF has a role in membrane repair, these data suggest a role for DYSF in the stability of the apical STB plasma membrane and may account, at least in part, for the increased shedding of microparticles from this membrane in PE. PMID- 19545897 TI - Differences of Ca2+ handling properties in identified central olfactory neurons of the antennal lobe. AB - Information processing in neurons depends on highly localized Ca2+ signals. The spatial and temporal dynamics of these signals are determined by a variety of cellular parameters including the calcium influx, calcium buffering and calcium extrusion. Our long-term goal is to better understand how intracellular Ca2+ dynamics are controlled and contribute to information processing in defined interneurons of the insect olfactory system. The latter has served as an excellent model to study general mechanisms of olfaction. Using patch-clamp recordings and fast optical imaging in combination with the 'added buffer approach', we analyzed the Ca2+ handling properties of different identified neuron types in Periplaneta americana's olfactory system. Our focus was on two types of local interneurons (LNs) with significant differences in intrinsic electrophysiological properties: (1) spiking LNs that generate 'normal' Na+ driven action potentials and (2) non-spiking LNs that do not express voltage activated Na+ channels. We found that the distinct electrophysiological properties from different types of central olfactory interneurons are strongly correlated with their cell specific calcium handling properties: non-spiking LNs, in which Ca2+ is the only cation that enters the cell to contribute to membrane depolarization, had the highest endogenous Ca2+ binding ratio and Ca2+ extrusion rate. PMID- 19545896 TI - Expression of aquaporin 1 and aquaporin 3 in fetal membranes and placenta in human term pregnancies with oligohydramnios. AB - OBJECTIVE: To explore the pathophysiology of oligohydramnios, the association between the expression of aquaporin 1 and aquaporin 3 in fetal membranes and placenta and oligohydramnios was investigated. METHODS: Sixty patients underwent elective cesarean sections at term were studied, 30 patients with isolated oligohydramnios and the other 30 with normal amniotic fluid volume (AFV). Real time polymerase chain reaction and immunohistochemistry were employed to determine expression and localization of aquaporin 1 and aquaporin 3 in amnion, chorion and placenta, respectively. RESULTS: The expression of aquaporin 1 and aquaporin 3 was detected in amnion, chorion and placenta using real-time RT-PCR. By immunohistochemistry, aquaporin 1 and aquaporin 3 protein expressions in amnion epithelia and chorion cytotrophoblasts were identified. In placenta, aquaporin 1 was detected in placental vessels, while aquaporin 3 was found in trophoblast cells. In comparison to normal AFV group, there was a significant decrease of aquaporin 1 expression in amnion in oligohydramnios group, but no significant difference in chorion and placenta between the two groups. The expression of the aquaporin 3 in amnion and chorion in oligohydramnios group was significantly decreased, while expression in placenta was significantly increased compared with that in normal AFV group. CONCLUSIONS: Alteration of aquaporin 1 and aquaporin 3 expression in fetal membranes and placenta may be important in the pathophysiology of isolated oligohydramnios. PMID- 19545898 TI - Molecular analysis of specificity of anti-nonylphenol polyethoxylate single-chain antibody fragments by grafting and designed point mutations. AB - Alkylphenol polyethoxylates and alkylphenols are widely distributed contaminants in the environment. Two anti-alkylphenol polyethoxylate monoclonal antibodies MOF3-139 and AP-14 were established to measure these chemicals by enzyme immunoassays in previous studies. Interestingly, these two monoclonal antibodies showed different specificity; AP-14 cross-reacts with nonylphenoxyacetic acid and nonylphenol, whereas MOF3-139 does not. To understand the molecular basis of the difference in specificity, single-chain Fv (scFv) antibodies derived from the monoclonal antibodies were each produced in Escherichia coli cells and characterized in competitive enzyme-linked immunosorbent assay. The scFv antibodies exhibited comparable reactivity profiles to the derived parent monoclonal antibodies. It was found that the VH domain of AP-14 play an important role in the cross-reaction when specificity tests were performed using variable domain-swapped scFv antibodies. An experiment using complementarity-determining region (CDR)-grafted scFv antibodies revealed that CDR1 and CDR2 of AP-14 are involved in the cross-reaction to nonylphenoxyacetic acid and nonylphenol, respectively. Site-directed mutagenesis was introduced in both regions and the assay revealed that 33rd Thr and 35th His in VH domain of AP-14 were highly involved in the cross-reaction with nonylphenoxyacetic acid and that 33rd Thr, 57th Asp, and 59th Glu were involved in the cross-reaction with nonylphenol. The findings herein would contribute to the antibody engineering for specificity modification and to the generation of an alkylphenol-specific recombinant antibody by antibody engineering. PMID- 19545899 TI - CCR7-specific migration to CCL19 and CCL21 is induced by PGE(2) stimulation in human monocytes: Involvement of EP(2)/EP(4) receptors activation. AB - The recent demonstration that newly recruited monocytes do not die at the site of inflammation, but migrate to draining lymph nodes, raises the question on the mechanism involved in this process. In this study, we demonstrate for the first time that prostaglandin E(2) (PGE(2)) regulates the expression and the activity of CCR7 in human blood-isolated monocytes as well as in the MONO-MAC-1 cell lineage. PGE(2) induces intracellular cAMP formation through engagement of the E prostanoid 2/E-prostanoid 4 (EP(2)/EP(4)) receptors present on monocytes. Migration to chemokines CCL19 and CCL21 in the PGE(2)-stimulated monocytes is mediated through the augmentation of cAMP concentration and furthermore, the cAMP/PKA pathway appears to act as the major inducer of CCR7 transcription in MONO-MAC-1. While p38 MAPK was induced by PGE(2), we observed that PGE(2) can downregulate p42/p44 MAPK phosphorylation. At the transcription level, inhibition of p38 MAPK inhibits CCR7 mRNA expression. Finally, we demonstrated that transcription factors CREB-1 and C/EBPalpha and C/EBPbeta are translocated to the nucleus following PGE(2) stimulation and bind the potent CCR7 promoter region. Our findings may have important implication for HIV-1 migration to the lymph nodes since macrophages and monocytes, particularly CD16 positive subset, are susceptible to HIV-1 infection. PMID- 19545900 TI - Primary intravitreal bevacizumab for diffuse diabetic macular edema: the Pan American Collaborative Retina Study Group at 24 months. AB - PURPOSE: To report the 24-month anatomic and Early Treatment Diabetic Retinopathy Study (ETDRS) best-corrected visual acuity (BCVA) response after primary intravitreal bevacizumab (Avastin; Genentech, Inc., San Francisco, CA; 1.25 or 2.5 mg) in patients with diffuse diabetic macular edema (DDME). In addition, a comparison of the 2 different doses of intravitreal bevacizumab (IVB) used is presented. DESIGN: Retrospective, multicenter, interventional, comparative case series. PARTICIPANTS: The clinical records of 115 consecutive patients (139 eyes) with DDME at 11 centers from 8 countries were reviewed. METHODS: Patients were treated with at least 1 intravitreal injection of 1.25 or 2.5 mg of bevacizumab. All patients were followed up for 24 months. Patients underwent ETDRS BCVA testing, ophthalmoscopic examination, optical coherence tomography (OCT), and fluorescein angiography (FA) at the baseline, 1-, 3-, 6-, 12-, and 24-month visits. MAIN OUTCOME MEASURES: Changes in BCVA and OCT results. RESULTS: The mean age of the patients was 59.4+/-11.1 years. The mean number of IVB injections per eye was 5.8 (range, 1-15 injections). In the 1.25-mg group at 1 month, BCVA improved from 20/150 (0.88 logarithm of the minimum angle of resolution [logMAR] units) to 20/107, 0.76 logMAR units (P<0.0001). The mean BCVA at 24 months was 20/75 (0.57 logMAR units; P<0.0001). Similar BCVA changes were observed in the 2.5-mg group: at 1 month, BCVA improved from 20/168 (0.92 logMAR units) to 20/118 (0.78 logMAR units; P = 0.02). The mean BCVA at 24 months was 20/114 (0.76 logMAR units; P<0.0001). In the 1.25-mg group, the mean central macular thickness (CMT) decreased from 466.5+/-145.2 microm at baseline to 332.2+/-129.6 microm at 1 month and 286.6+/-81.5 microm at 24 months (P<0.0001). Similar results were obtained in the 2.5-mg group. CONCLUSIONS: Primary IVB at doses of 1.25 to 2.5 mg seem to provide stability or improvement in BCVA, OCT, and FA in DDME at 24 months. The results show no evident difference between IVB at doses of 1.25 or 2.5 mg. PMID- 19545901 TI - Intravitreal bevacizumab for refractory pseudophakic cystoid macular edema: the Pan-American Collaborative Retina Study Group results. AB - OBJECTIVE: To determine the feasibility, safety, and clinical effect of intravitreal (IVT) bevacizumab (Avastin; Genentech, Inc., San Francisco, CA) in patients with refractory cystoid macular edema (CME) after cataract surgery. DESIGN: Interventional, retrospective, multicenter study. PARTICIPANTS: Thirty six eyes of 31 patients with refractory CME after cataract surgery and with a mean age of 68.2 years (range, 67-87 years). METHODS: Patients were treated with at least 1 IVT injection of 1.25 or 2.5 mg bevacizumab. Patients were followed up for 12 months. MAIN OUTCOME MEASURES: Best-corrected visual acuity (BCVA) and central macular thickness (CMT) by optical coherence tomography (OCT). RESULTS: Twenty-six eyes (72.2%) demonstrated improvement of BCVA (> or =2 Early Treatment Diabetic Retinopathy Study [ETDRS] lines), and no eye experienced worsening of visual acuity (> or =2 ETDRS lines). Mean baseline BCVA was 20/200 (0.96 logarithm of the minimum angle of resolution [logMAR] units), and the mean 12 month BCVA was 20/80 (0.62 logMAR units; P<0.0001). Optical coherence tomography demonstrated that mean CMT at baseline was 499.9 microm (range, 298-784 microm) and decreased to a mean of 286.1 microm (range, 168-499 microm) at 12 months (P<0.0001). Four (11%) eyes received 2 injections, 10 (27.8%) eyes received 3 injections, 10 (27.8%) eyes received 4 injections, 1 (2.8%) eye received 5 injections, and 1 (2.8%) eye received 6 injections. The mean number of injections was 2.7 (range, 1-6), and the mean interval between injections was 15.1 weeks (range, 4-45 weeks). No ocular or systemic adverse events were observed. CONCLUSIONS: Short-term results suggest that IVT bevacizumab is well tolerated in patients with refractory pseudophakic CME. Treated eyes had a significant improvement in BCVA and decrease in macular thickness by OCT at 12 months. PMID- 19545902 TI - Robot-assisted vitreoretinal surgery: development of a prototype and feasibility studies in an animal model. AB - PURPOSE: To develop a prototype robotic system designed to assist vitreoretinal surgery and to evaluate its accuracy and maneuverability. DESIGN: Experimental study. PARTICIPANTS: This study used harvested porcine eyes. METHODS: After development of a prototype robotic system, pointing accuracy tests of the system were performed on graph paper and in harvested porcine eyes. The average maximal deviation from the aiming point to the actual position of the tip of the instrument was compared between manually conducted procedures and those conducted with robotic assistance. The feasibility of creating posterior vitreous detachment (PVD), retinal vessel sheathotomy (RVS), and retinal vessel microcannulation also were evaluated in porcine eye models, and the success rates of 4 consecutive attempts for each kind of procedure were evaluated. MAIN OUTCOME MEASURES: The average maximum deviation in pointing accuracy tests both on graph paper and in animal eye models was a main outcome measure. The success rate of making PVD, RVS, and retinal vessel microcannulation was the other primary outcome measure. RESULTS: The pointing accuracy was superior with robotic assistance both on graph paper (327.0 microm vs. 32.3 microm) and in animal eye models (140.8 microm vs. 33.5 microm). Creating PVD, RVS, and retinal vessel microcannulation was feasible in 4 of 4 attempts, 4 of 4 attempts, and 2 of 4 attempts, respectively. The 2 failures in microcannulation were considered to be the result of difficulty in visual differentiation between the retinal vessel and retina in harvested porcine eyes. CONCLUSIONS: Improved accuracy and desirable feasibility of a prototype robotic system to assist vitreoretinal surgery were shown in this study. Research for wider implementation of robot-assisted surgery should be continued; there are some hurdles to overcome. PMID- 19545903 TI - Bilateral acute depigmentation of the iris: report of 26 new cases and four-year follow-up of two patients. AB - PURPOSE: To report new cases of bilateral acute depigmentation of the iris (BADI), a recently described clinical entity, and to report the 4-year follow-up of 2 patients that was published previously. DESIGN: A retrospective case series. PARTICIPANTS: Twenty-six Turkish patients who were diagnosed with BADI between 2006 and 2008 and 2 patients who were reported previously. METHODS: We reviewed the patients' charts and clinical photographs. Observation procedures included clinical examination, anterior segment color photography, laser flare photometry, and pupillometry. We performed an anterior chamber tap in 2 patients for polymerase chain reaction (PCR) to demonstrate the DNA of herpes simplex virus (HSV), varicella zoster virus (VZV), and cytomegalovirus (CMV) in the aqueous samples. MAIN OUTCOME MEASURES: Demographic features, presenting symptoms, laboratory findings, changes in iris stromal pigment and architecture, and time to resolution of pigment dispersion in the anterior chamber. RESULTS: Nineteen patients were female, and 7 patients were male. Mean age was 32.3+/-8.6 years. All had bilateral involvement. Twenty patients (76.9%) presented with photophobia and red eyes, and 4 patients (15.4%) presented with a recent change in eye color. Ten patients (38.5%) had flu-like symptoms preceding the onset of ocular symptoms. Diagnostic laboratory workup, viral serology, and PCR analysis of the aqueous humor were unrewarding. Diffuse depigmentation of the iris stroma from the collarette to the iris root was seen in 16 patients, and geographic areas of depigmentation were seen in 10 patients. There was heavy pigment deposition in the trabecular meshwork in all patients. Anterior chamber flare was elevated in eyes with circulating pigment. The pupil was not affected. Twenty patients received topical corticosteroids. Pigment dispersion resolved in 1 to 16 weeks (median, 9 weeks). The intraocular pressure was elevated in 8 steroid-treated eyes but was controlled with antiglaucomatous medications. In 2 patients reported previously, the depigmented iris stroma became repigmented after 4 years. CONCLUSIONS: Patients with BADI present with bilateral, symmetrical, nontransilluminating depigmentation of the iris stroma and pigment discharge into the anterior chamber. Young female persons are more commonly affected. The cause remains unknown. After 4 years, the ocular findings in 2 patients normalized. PMID- 19545904 TI - Lack of correlation between the histologic and magnetic resonance imaging results of optic nerve involvement in eyes primarily enucleated for retinoblastoma. AB - PURPOSE: To correlate the histologic and magnetic resonance imaging results of the optic nerve in eyes primarily enucleated for retinoblastoma. DESIGN: Retrospective, clinicopathologic correlation. PARTICIPANTS: Sixty-seven consecutive patients with retinoblastoma who underwent primary enucleation. METHODS: The histologic results of 67 eyes from 67 patients with retinoblastoma who underwent primary enucleation between March 1997 and January 2008 were studied for evidence of optic nerve invasion. Two neuroradiologists independently reviewed available preoperative magnetic resonance imaging studies with special emphasis on nonenhanced T2-weighted and gadolinium-enhanced T1-weighted imaging for evidence of optic nerve invasion. A weighted kappa statistic was used to assess agreement between observers. MAIN OUTCOME MEASURES: Correlation between neuroradiologists and histologic results. RESULTS: Of the 67 eyes studied, 60 had preoperative magnetic resonance images, 58 of which were deemed appropriate for review by both neuroradiologists. Review of the histologic results showed optic nerve involvement in 62 (93%) of 67 eyes: 28 prelaminar (42%), 24 laminar (36%), and 10 postlaminar (15%). On review of the magnetic resonance scans, the first neuroradiologist identified optic nerve involvement in 57 (95%) of 60 eyes: 26 prelaminar (43%), 10 laminar (17%), and 11 postlaminar (18%). The second neuroradiologist identified optic nerve involvement in 46 (77%) of 60 eyes: 33 prelaminar (55%), 9 laminar (15%), and 4 postlaminar (7%). Moderate agreement existed between neuroradiologists (kappa, 0.55). Poor and fair agreement existed between each of the 2 neuroradiologists and histologic results, respectively (kappa, 0.29 and 0.17). Exophytic tumors showed the greatest disparity (kappa, 0.20 and -0.13) between magnetic resonance imaging and histologic results. CONCLUSIONS: Limited correlation was found between magnetic resonance imaging and histologic results in assessing optic nerve invasion in eyes with retinoblastoma. Magnetic resonance imaging using routine imaging technologies, although useful in the evaluation of retinoblastoma, has limited usefulness in assessing the exact extent of optic nerve invasion; high-risk features of retinoblastoma such as postlaminar invasion remain best defined by histologic analysis. This study demonstrates that the interpretation of optic nerve involvement by a radiologist should not be the determining factor to defer enucleation in favor of neoadjuvant therapy. PMID- 19545905 TI - Does open access in ophthalmology affect how articles are subsequently cited in research? AB - OBJECTIVE: To determine whether the concept of open access affects how articles are cited in the field of ophthalmology. DESIGN: Type of meta-analysis. PARTICIPANTS: Examination of 480 articles in ophthalmology in the experimental protocol and 415 articles in the control protocol. METHODS: Four subject areas were chosen to search the ophthalmology literature in the PubMed database using the terms "cataract," "diabetic retinopathy," "glaucoma," and "refractive errors." Searching started in December of 2003 and worked back in time to the beginning of the year. The number of subsequent citations for equal numbers of both open access and closed access (by subscription) articles was quantified using the Scopus database and Google search engine. Number of authors, article type, country/region in which the article was published, language, and funding data were also collected for each article. A control protocol was also carried out to ascertain that the sampling method was not systematically biased by matching 6 ophthalmology journals (3 open access, 3 closed access) using their impact factors, and employing the same search methodology to sample open access and closed access articles. MAIN OUTCOME MEASURES: Number of citations. RESULTS: The total number of citations was significantly higher for open access articles compared to closed access articles for Scopus (mean 15.2 versus 11.5, P < .0005, Mann-Whitney U = 20029, and Google (mean 6.4 versus 4.0, P < .0005, Mann-Whitney U = 21281). However, univariate general linear model (GLM) analysis showed that access was not a significant factor that explained the citation data. Author number, country/region of publication, subject area, language, and funding were the variables that had the most effect and were statistically significant. Control protocol results showed no significant difference between open and closed access articles in regard to number of citations found by Scopus: open access: mean = 17.8; SD (standard deviation) = 23.70; closed access: mean = 19.1; SD = 20.31; Mann-Whitney test, P = 0.730, Mann-Whitney U = 20584. CONCLUSIONS: Unlike other fields of science, open access thus far has not affected how ophthalmology articles are cited in the literature. PMID- 19545906 TI - Regulatory T cells in prenatal blood samples: variability with pet exposure and sensitization. AB - Fetal exposures have come under investigation as risk factors of early life allergic disease. In this study we aimed to examine the relationships between dog or cat exposure and naturally occurring regulatory T cells (Treg cells), thought to play an important role in immune tolerance, in pregnant women. A cross sectional analysis was conducted among 204 pregnant women who were queried regarding dog and cat exposure. Treg cells (CD4+CD25+Foxp3+ lymphocytes) and allergen-specific IgE were measured in venous blood samples. Atopy was defined as allergen-specific IgE > or =0.35kU/l reactive with common allergens including dust mite, dog, cat, Timothy grass, ragweed, Alternaria alternata, egg white or cockroach. Nonparametric Wilcoxon rank sum tests and linear regression models of log transformed Treg cell levels were used in analyses. Among women sensitized to dog, those who had a dog or cat in the home had lower Treg cell levels compared with those who had no dog or cat. However, among women not sensitized to dog, those with a dog or cat in the home had higher Treg cell levels compared with those who did not. Among women sensitized to cat, those who had a dog or cat in the home had lower Treg cell levels compared with those who had no dog or cat. Gestational age at blood draw did not affect the associations. We conclude that Treg cell levels during pregnancy vary in association with both dog and cat exposure and atopic status. PMID- 19545907 TI - Does reward frequency or magnitude drive reinforcement-learning in attention deficit/hyperactivity disorder? AB - Children with attention-deficit/hyperactivity disorder (ADHD) show an impaired ability to use feedback in the context of learning. A stimulus-response learning task was used to investigate whether (1) children with ADHD displayed flatter learning curves, (2) reinforcement-learning in ADHD was sensitive to either reward frequency, magnitude, or both, and (3) altered sensitivity to reward was specific to ADHD or would co-occur in a group of children with autism spectrum disorder (ASD). Performance of 23 boys with ADHD was compared with that of 30 normal controls (NCs) and 21 boys with ASD, all aged 8-12. Rewards were delivered contingent on performance and varied both in frequency (low, high) and magnitude (small, large). The findings showed that, although learning rates were comparable across groups, both clinical groups committed more errors than NCs. In contrast to the NC boys, boys with ADHD were unaffected by frequency and magnitude of reward. The NC group and, to some extent, the ASD group showed improved performance, when rewards were delivered infrequently versus frequently. Children with ADHD as well as children with ASD displayed difficulties in stimulus response coupling that were independent of motivational modulations. Possibly, these deficits are related to abnormal reinforcement expectancy. PMID- 19545909 TI - Pathological gamblers may just be less attentive, not necessarily more impulsive. PMID- 19545910 TI - Molecular characterisation of porcine Forkhead-box p3 (Foxp3). AB - In swine the phenotypical identification of regulatory T cells (Tregs) was limited so far to the surface expression of CD4 and CD25. However, with the discovery of the Treg-specific transcription factor forkhead-box p3 (Foxp3) in mice and humans a powerful marker for the identification of Tregs is available. Recently, we published data on a murine anti-mouse/rat Foxp3 antibody (FJK-16s) showing cross-reactivity with the putative porcine Foxp3 protein in lymphoid cells but the final proof for the specific cross-reactivity of this antibody was missing. By performing RACE-experiments, we have sequenced the entire porcine Foxp3 cDNA which is 1296 nucleotides in length and codes for a polypeptide of 432 amino acids. The porcine Foxp3 nucleotide and amino acid sequences show high homology to all known orthologues from other mammals, with the greatest homology with the bovine sequence. To demonstrate the specificity of the FJK-16s antibody for the porcine Foxp3 protein, HEK293T cells were transfected with porcine Foxp3 containing the FJK-16s-specific binding region and the expression of the epitope was identified by immuno-staining. In conclusion, this study represents the final proof for the specificity of the murine FJK-16s antibody for the porcine Foxp3 homologue and therefore strengthens future work on porcine Tregs. PMID- 19545913 TI - Surgical management of cholesteatoma: the two main options and the third way- atticotomy/limited mastoidectomy. AB - Chronic otitis media with cholesteatoma is considered an "unsafe" ear and generally requires surgical management. This is particularly challenging in children due to anatomical, pathophysiological and social reasons. There are different approaches for this objective. The two main options are the canal wall up and canal wall down mastoidectomy. The aim of this article is to compare the advantages and disadvantages of canal wall up and canal wall down method and present the third way of surgical management: the inside-outside approach through an endaural incision. This technique includes atticotomy, atticoantrostomy or mastoidectomy (mostly very limited) according to the size and location of the cholesteatoma. This technique contributes to the successful surgical management of cholesteatoma, eradicating the disease with the creation of small, dry, self cleaning cavities and no pinna protrusion. Moreover, there is no need for meatoplasty or obliteration. However, we should never forget that in ear surgery the choice of the operative procedure should take into account the needs of the patient, the extent of the disease, and the surgeon's experience. PMID- 19545912 TI - Ischemic preconditioning-induced neuroprotection is associated with differential expression of IL-1beta and IL-1 receptor antagonist in the ischemic cortex. AB - Ischemic preconditioning (IP) is a phenomenon that organs develop a tolerance toward subsequent lethal ischemic insults. Among the factors that are involved in IP, IL-1beta and its endogenous receptor antagonist IL-1ra have been identified as important players in the induction of IP. The present study investigated whether IP affects the levels of these two antagonistic proteins during tolerance and reperfusion periods after ischemic stroke. The IP 24 h prior to ischemic stroke resulted in neuroprotection in the cortex. IP-induced protection is accompanied by increased IL-1beta gene and IL-1ra gene and protein levels during the tolerance period. In the post-ischemic cortex, IP resulted in the suppression of IL-1beta mRNA and protein levels at 6 h without affecting IL-1ra expression and the up-regulation of IL-1ra protein at 24 h. These findings demonstrate that IP differentially regulates cortical IL-1beta and IL-1ra expression before and after ischemic stroke and suggest that the shift toward an anti-inflammatory state in the post-ischemic cortex may contribute to IP-induced neuroprotection. PMID- 19545911 TI - Immunodominant epitope and properties of pyroglutamate-modified Abeta-specific antibodies produced in rabbits. AB - N-truncated and N-modified forms of amyloid beta (Abeta) peptide are found in diffused and dense core plaques in Alzheimer's disease (AD) and Down's syndrome patients as well as transgenic mouse models of AD. Although the pathological significance of these shortened forms Abeta is not completely understood, previous studies have demonstrated that these peptides are significantly more resistant to degradation, aggregate more rapidly in vitro and exhibit similar or, in some cases, increased toxicity in hippocampal neuronal cultures compared to the full length peptides. In the present study we further investigated the mechanisms of toxicity of one of the most abundant N-truncated/modified Abeta peptide bearing amino-terminal pyroglutamate at position 3 (AbetaN3(pE)). We demonstrated that AbetaN3(pE) oligomers induce phosphatidyl serine externalization and membrane damage in SH-SY5Y cells. Also, we produced AbetaN3(pE)-specific polyclonal antibodies in rabbit and identified an immunodominant epitope recognized by anti-AbetaN3(pE) antibodies. Our results are important for developing new immunotherapeutic compounds specifically targeting AbetaN3(pE) aggregates since the most commonly used immunogens in the majority of vaccines for AD have been shown to induce antibodies that recognize the N terminal immunodominant epitope (EFRH) of the full length Abeta, which is absent in N-amino truncated peptides. PMID- 19545914 TI - siRNA delivery using peptide transduction domains. AB - Targeting mRNA degradation by short interfering RNAs (siRNAs) offers great potential to treat multiple diseases. However, owing to their high molecule size and strong anionic charge, siRNAs cannot pass through the highly regulated and restricted plasma membrane. To overcome these problems, many approaches have been developed and applied in clinical trial. However, other siRNA delivery systems are still required to enhance the siRNA uptake. Over the past 20 years, peptide transduction domains (PTDs) have been discovered that can cross the cellular membrane by themselves despite their high molecule size. PTDs have been used for the delivery of a wide range of molecules including peptides, proteins and antisense oligonucleotides and applied in vivo systems. Here, we review siRNA delivery using PTDs in vitro and in vivo and discuss its potential for use in siRNA-based therapy. PMID- 19545915 TI - The copper-induced reduction of critical swimming speed in rainbow trout (Oncorhynchus mykiss) is not caused by changes in gill structure. AB - In the present experiments, we investigated, if copper exposure causes a decrease in the critical swimming speed, as has been observed earlier, and if such a decrease can be explained by a disturbance in gill structure. The results show that this is not the case. While copper exposure tended to increase the thickness of gill diffusion barrier in resting fish, a similar difference was not seen in swimming animals. However, copper exposure caused a decrease in the critical swimming speed. This decrease must thus be caused by factors other than structural changes in the gills, associated with copper exposure. PMID- 19545916 TI - Subclinical hypothyroidism (SCH) is not associated with metabolic derangement, cognitive impairment, depression or poor quality of life (QoL) in elderly subjects. AB - The objective of this study was to examine the relationship between SCH and metabolic and neuropsychiatric derangements in Korean elderly subjects. Nine hundred and eighteen euthyroid and SCH subjects aged 65 years and older were enrolled in the present study. We compared anthropometric (age, sex, blood pressure, body mass index=BMI, body fat contents=BF, waist circumference=WC and hip circumference=HC), laboratory (glucose and lipid profile, renal and liver functions, and C-reactive protein=CRP levels), and neuropsychiatric (neuropsychological function, mood, and QoL) parameters of SCH subjects with those of euthyroid subjects. Associated metabolic disorders of the subjects were also compared between the euthyroid and SCH groups. We could not observe any differences between the SCH group and the euthyroid group with regards to metabolic parameters as well as neuropsychiatric parameters. These findings may suggest that SCH does not appear to be associated with metabolic and neuropsychiatric derangement in elderly subjects. PMID- 19545917 TI - Developing a self-reported comorbidity index to predict mortality of community dwelling older adults. AB - Current common comorbidity measures have poor to moderate predictive validity of mortality of community-dwelling older adults. Hence, our aim is to develop a simpler resource-efficient self-reported comorbidity index in the prediction of survival. 113 older adults in Greater Manchester, United Kingdom attended a routine medical examination whereby information gathered was used to construct Charlson Comorbidity Index (CCI). They completed the Cornell Medical Index (CMI) questionnaire and reported the number of medication prescribed to them. We compared the ability of CCI, CMI, number of medication, age and sex to predict mortality of the sample over 7-year period using Cox-regression and Kaplan-Meier plot and rank test. None of the variables individually was significant when tested using either Cox-regression via ENTER method or Kaplan-Meier test. Remarkably, by means of forward step-wise Cox-regression, two variables emerged significant: (i) number of medicine (beta coefficient=0.229, SE=0.090 and p=0.011) and (ii) age (beta coefficient=0.106, SE=0.051 and p=0.037). We demonstrated that simple count of medication predicted mortality of community dwelling older adults over the next 7 years more accurately than CMI or CCI. Further works involving a larger scale of subjects is needed for use in epidemiological study of survival where cost and resources are concerned. PMID- 19545918 TI - Decline of physical and cognitive conditions in the elderly measured through the functional reach test and the mini-mental state examination. AB - There are several tests used to evaluate the psychophysical characteristics of the elderly and, of these, the most suitable are the functional reach (FR) test, an index of the aptitude to maintain balance in an upright position, and the mini mental state examination (MMSE), a global index of cognitive abilities. The sample of elderly people we analyzed involved 50 healthy subjects divided into three age-groups (15 subjects from 55 to 64 years, 19 from 65 to 74 years, and 16 over 75 years of age); they underwent an FR test, which consists first in the measurement of the anthropometric characteristics, then in the execution of the test itself, and finally in the study of the upright posture by the analysis of the center of pressure (COP) trend; they underwent an MMSE as well to evaluate the main areas of the cognitive function concerning space-time orientation, short term memory, attention ability, calculation ability and constructive praxis. The results of these tests show, according to the age of the subject, a loss of physical performance (FR, FR related to height, and COP displacement), as well as a loss of cognitive abilities; however, in all cases the only significant changes are those between the first and the other two age-groups. Finally, a comparison between FR and MMSE shows a more rapid decline of physical performance compared to cognitive performance. PMID- 19545919 TI - Interrupted aortic arch associated with aortopulmonary window in a 20 year-old young adult. AB - Interrupted aortic arch (IAA) and aortopulmonary window (APW) are rare but well described congenital malformations. However, coexistence of these anomalies in young adults is an unusual event. Patients rarely reach adulthood without surgical correction in the neonatal period. The authors describe a 20-year-old patient, who presented with exertional dyspnea and was diagnosed as having IAA and concomitant APW. IAA just distal to the left subclavian artery and a large APW were demonstrated by chest multi-slice computed tomography angiography and cardiac catheterization. PMID- 19545920 TI - Percutaneous retrieval of an embolized pacemaker lead from the pulmonary artery. AB - Migration of retained fractured pacemaker electrodes to the pulmonary artery is a rare but recognised complication. It is imperative that if this occurs the electrode is removed from this position due to the increased risk of associated infection and thrombus formation. Historically open surgery has been required to manage this potentially serious complication, however as illustrated by this case, a percutaneous approach can be employed successfully. PMID- 19545921 TI - Atrial electrical remodeling in a canine model of sinus node dysfunction. AB - AIMS: To study atrial tachycardia-induced electrical remodeling in a canine model of sinus node dysfunction (SND). MATERIALS AND METHODS: A canine model of SND was established by contacting a cotton patch with 20% formaldehyde on the sinus node. Atrial effective refractory period (ERP), ERP dispersion, and inducibility of atrial fibrillation (AF) were recorded at multiple sites in the atrium, before and after SND induction as well as after rapid atrial pacing. The recovery of atrial ERP in the left and right atrium (LA and RA) after cessation of atrial pacing was also recorded. RESULTS: Compared with baseline, the atrial ERPs were shortened after SND (P<0.05). After rapid atrial stimulation, the atrial ERPs were further decreased significantly (P<0.05), and the dispersion of atrial ERPs measured at different pacing cycle lengths (PCLs) showed significant variation. Seven sites were used to induce AF in each dog (56 sites in 8 dogs). The average duration and inducibility of AF after SND was increased compared with baseline (16.5+/-4.7 vs 2.3+/-1.2 s and 12/56 vs 4/56 sites, P<0.05). After rapid atrial stimulation, the average duration and inducibility of AF were further increased (16.5+/-4.7 vs 33.6+/-16.1 s and 12/56 vs 25/56 sites, P<0.05). The recovery of atrial ERP in LA was significantly delayed compared to the RA. CONCLUSION: SND induces atrial electrical remodeling which is further aggravated by atrial tachycardia. Therefore, SND creates an electrophysiological substrate that facilitates AF initiation and perpetuation. PMID- 19545922 TI - Interatrial conduction in patients with paroxysmal atrial fibrillation and in healthy subjects. AB - BACKGROUND: Altered atrial conduction is linked to susceptibility to atrial fibrillation (AF). Whether signal propagation to left atrium (LA) during sinus rhythm differs between patients with paroxysmal lone AF and healthy subjects is not known. METHODS: In 107 patients with lone paroxysmal AF (age 45+/-12 years) and 94 controls 99-channel magnetocardiography (MCG) was recorded over anterior chest. The duration of the atrial wave (Pd) and the MCG maps over time intervals corresponding to early and later LA activations were determined. Based on magnetic field orientations in LA maps, MCG atrial waves were classified into 3 types which are related to distinct interatrial conduction routes: Type 1 to Bachmann bundle, Type 2 to margin of fossa ovalis or multisite, and Type 3 to coronary sinus ostial connections. RESULTS: Pd was longer in AF patients than in controls (112+/-13 vs. 104+/-13; p<0.001), which was most obvious in Type 1 wave (109+/-12 vs. 102+/-11 ms, p=0.003). The distribution of the atrial wave types differed between AF patients and controls: Type 1 occurred in 67% and Type 2 in 20% of controls whereas Type 1 occurred in 54% and Type 2 in 42% of AF patients, p<0.01 for difference. CONCLUSIONS: Susceptibility to paroxysmal lone AF is associated with propagation of atrial signal to LA via margin of fossa ovalis or multiple pathways. When conduction occurs via Bachmann bundle, it is related with prolonged atrial activation. Thus altered and alternative conduction pathways may contribute to pathogenesis of lone AF. PMID- 19545923 TI - A cross-sectional study of the prevalence of foot lesions in post-weaning pigs and risks associated with floor type on commercial farms in England. AB - We carried out a cross-sectional study during 2003 and 2004 to establish the prevalence and risk factors associated with floor type for commonly observed foot lesions in pigs aged 6, 8 and 14 weeks. The overall prevalence of foot lesions was 39.6% in 2283 pigs from 90 representative pig farms in England. The most prevalent lesions were heel/sole bruising (7.1%), heel/sole erosion (10.8%), heel flaps (8.4%) and toe erosion (11.6%). Pigs were kept on either solid (41%), partly slatted (28%) or fully slatted (31%) floors. Of the 104 pens with a solid floor, 26% of pens were outside with straw bedding on a soil base, 33% were indoors with deep bedding on solid concrete, 25% were partly deeply bedded on solid concrete and 16% were sparsely bedded on solid concrete. Only six of the pens with partly slatted floors were bedded. Multilevel logistic-regression models were built using data from 100 farms to examine the risks for individual foot lesions with prevalences >5%. The prevalence of toe erosions was positively associated with deep bedding, whereas deep bedding and soil floors were negatively associated with the prevalence of heel/sole erosions. Heel flaps and heel/sole bruising were both associated with slatted floors, possibly indicating a common aetiology. The greatest reduction in prevalence of all these lesions, from AFp calculations, would be achieved by moving pigs from slatted floors onto solid floors. PMID- 19545924 TI - Diversity and dynamics of the microbial community during the manufacture of Calenzana, an artisanal Corsican cheese. AB - We studied the diversity and dynamics of the microbiota of Calenzana, a Corsican raw milk cheese by microbial counting and culture-independent methods (TTGE and DGGE). Cheese from two farms, one producing goat cheese and the other one sheep cheese, was studied. The usual process for cheese making, without starter adjunct, was used. Lactococci and mesophilic lactobacilli were the dominant components of the flora during the early stages of the process. Microbial counting showed that the populations of salt-tolerant bacteria, yeasts and moulds were lower than in other artisanal Corsican cheeses. This difference was probably due to the surface microflora being removed during ripening. TTGE indicated that Lactococcus lactis ssp. lactis was the dominant subspecies throughout the process of Calenzana cheese making. DGGE showed the presence of numerous surface bacteria, (coryneforms) and various Gram-negative bacteria. Relationships between physico-chemical characteristics of the cheese and microflora change were also established. For example, the high NaCl content may explain the decrease of the lactic acid bacterial population during ripening. This study shows the consequences of various technological parameters on the diversity and dynamics of dairy microbial community. PMID- 19545925 TI - Low-dose pioglitazone increases serum high molecular weight adiponectin and improves glycemic control in Japanese patients with poorly controlled type 2 diabetes. AB - We investigated the effects of low-dose pioglitazone (7.5mg/day) on serum high molecular weight (HMW) adiponectin and fluid retention (estimated from hematocrit) in 14 male and 16 female patients with type 2 diabetes. All of them were being treated with sulfonylureas and had poor glycemic control. Patients were given 7.5 mg/day of pioglitazone and were followed for 12 weeks at monthly intervals. In all 30 patients, HbA1c was significantly decreased after 12 weeks of treatment with pioglitazone (8.2+/-0.7% vs. 7.4+/-0.8%, P<0.0001). Serum HMW adiponectin increased markedly from 5.2 (2.4, 8.6) microg/ml at baseline to 9.8 (4.1, 12.6) microg/ml at the end of pioglitazone treatment (P<0.0001). When the changes were evaluated separately for each sex, diabetic men showed no increase of body weight or BMI after treatment, while HbA1c decreased significantly, and did Hct. Serum HMW adiponectin increased significantly after treatment. In diabetic women, neither body weight nor BMI increased after treatment with pioglitazone, as was the case for the men. HbA1c decreased significantly, and did Hct. Serum HMW adiponectin increased significantly after treatment. In conclusion, low-dose pioglitazone therapy could significantly improved glycemic control and markedly increased serum HMW adiponectin in both male and female Japanese patients with type 2 diabetes. PMID- 19545927 TI - On the role of platelets in the pathogenesis of viral hepatitis. PMID- 19545926 TI - Hepatitis C virus-induced hepatocarcinogenesis. AB - Although there is strong evidence that hepatitis C virus (HCV) is one of the leading causes of hepatocellular carcinoma (HCC), there is still much to understand regarding the mechanism of HCV-induced transformation. While liver fibrosis resulting from long-lasting chronic inflammation and liver regeneration resulting from immune-mediated cell death are likely factors that contribute to the development of HCC, the direct role of HCV proteins remains to be determined. In vitro studies have shown that HCV expression may interfere with cellular functions that are important for cell differentiation and cell growth. However, most studies were performed in artificial models which can only give clues for potential mechanisms that need to be confirmed in more relevant models. Furthermore, the difficulty to identify HCV proteins and infected liver cells in patients, contributes to the complexity of our current understanding. For these reasons, there is currently very little experimental evidence for a direct oncogenic role of HCV. Further studies are warranted to clarify these issues. PMID- 19545928 TI - Clinical significance of cancer/testis antigens expression in patients with non small cell lung cancer. AB - Cancer/testis antigens (CT antigens) are thought to be suitable targets for antigen-specific immunotherapy, because of the cancer-specific expression except for the testis among various normal tissues and no-expression of HLA class I in the testis. In the present study, the expressions of CT antigens (MAGE-A3, MAGE A4, NY-ESO-1 and KK-LC-1) in non-small cell lung cancer (NSCLC) were analyzed by RT-PCR. The subjects were 239 patients with NSCLC who underwent surgery from 2001 to 2005 in our department. The expression rates of MAGE-A3, MAGE-A4, NY-ESO-1 and KK-LC-1 were 23.8%, 20.1%, 10.5% and 32.6% in patients with NSCLC, respectively. MAGE-A4 was expressed more frequently in male (25.3%) than in female (10.6%) (p<0.01). The positive proportion of MAGE-A4 was higher in stages II-IV (30.6%) than in stage I (12.8%) (p<0.01). Both of MAGE-A3 and MAGE-A4 were expressed more frequently in squamous cell carcinoma than in adenocarcinoma (p<0.01). Such tendency was not observed among NY-ESO-1 and KK-LC-1 expression. KK-LC-1 was expressed in 32.1% of patients with adenocarcinoma and in 36.5% of patients with squamous cell carcinoma. Patients with positive MAGE-A4 expression showed significantly poorer overall survival than those without MAGE-A4 expression (p=0.013), and such effect on survival was also observed, when the analysis was limited to patients at stage I (p=0.0037). Expression of MAGE-A3, NY-ESO-1 or KK LC-1 did not affect survival of patients with NSCLC significantly, however, expression of at least one of such CT antigens negatively affect survival of patients with NSCLC (p=0.045). PMID- 19545929 TI - Gemcitabine combined with either pemetrexed or paclitaxel in the treatment of advanced non-small cell lung cancer: a randomized phase II SICOG trial. AB - PURPOSE: To estimate the safety, activity, and impact on quality of life of a combination of gemcitabine and pemetrexed in patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) in the context of a randomized two stage phase II study. PATIENTS AND METHODS: Patients in stage IIIB or IV NSCLC were randomly allocated to receive either gemcitabine 1250 mg/m(2) on day 1, and pemetrexed (Alimta) 500 mg/m(2) followed by gemcitabine 1250 mg/m(2) on day 8 of a 3-weekly cycle (GA arm), or paclitaxel 120 mg/m(2) followed by gemcitabine 1000 mg/m(2), both given on days 1 and 8 of a 3-weekly cycle (PG arm). RESULTS: 105 (GA arm, 51; PG arm, 54) eligible patients (stage IV, 32 and 30, respectively) were enrolled into this study; thereafter, accrual was stopped due to first-stage analysis. The response rate was 20% (95% confidence interval [CI], 10-33%) in the GA arm, and 32% (95% CI, 20-46%) in the PG arm. Median progression-free survival was 5.1 (95% CI, 3.7-6.5) months in the GA arm, and 8.3 (95% CI, 5.9-10.7) months in the PG arm, while median overall survival was 10.5 (95% CI 7.1-13.9), and 13.3 (95% CI 11.7-14.9) months, respectively. Severe neutropenia (36% vs 22%), and febrile neutropenia (14% vs 7%) were more common with the GA regimen, while hair loss (52% vs 16%) and any grade peripheral neuropathy (31% vs 2%) occurred more frequently with PG regimen. Other severe side effects of GA regimen were diarrhoea (10%), liver enzyme derangement (10%), and fatigue (8%). CONCLUSION: The GA regimen was tolerated and moderately active in advanced or metastatic NSCLC. However, this combination did not yield any advantage in comparison with the PG regimen, and does not deserve further evaluation. PMID- 19545930 TI - Lung cancer risk and cigarette smoking, lung tuberculosis according to histologic type and gender in a population based case-control study. AB - We examined whether the strength of the association of cigarette smoking differs according to histological type and gender, and assessed other risk factors, in particular, tuberculosis. We recruited cases from the Korean Academy of Tuberculosis and Respiratory Diseases and controls from Chungju, a local site of the Korean Multi-Center Cancer Cohort. We matched one case to one control for females and two cases to one control for males according to age (or=70 years old). We used unconditional logistic regression to calculate the odds ratio (OR) and 95% confidence interval (CI) to estimate lung cancer risk by histologic type for males and females separately. The OR (95% CI) of 40 or more pack-years smoked relative to never smokers was 6.78 (4.17-11.00), 3.49 (1.83 6.33), and 2.72 (1.57-4.72) for males, and 13.72 (3.23-58.18), 12.18 (3.12 47.57), and 7.11 (1.78-28.43) for females for squamous cell, adenocarcinoma, and small cell carcinoma, respectively. Among males, the respective OR (95% CI) for past and current history of lung tuberculosis was 3.21 (2.12-4.90), 2.69 (1.63 4.45), and 1.52 (0.83-2.78), and for females was 2.40 (1.30-4.42), 4.20 (2.75 6.39), and 1.37 (0.61-3.06). Our findings provide additional evidence that women are more susceptible to the carcinogenic effects of tobacco, smoking has a higher risk for squamous cell and small cell carcinoma than adenocarcinoma, and tuberculosis is a potential risk factor for certain lung cancer histologic types. PMID- 19545931 TI - The karyopherin CRM1 is required for dendritic cell maturation. AB - Dendritic cells (DC) are the most potent antigen-presenting cells (APC) of the immune system and are specialized to activate T as well as B cell-dependent immune responses. Mature DC are characterized by expression of CD83, a surface molecule that has been postulated to be required for efficient DC activity. Here we show that Leptomycin B (LMB), a highly specific inhibitor of the nuclear export receptor CRM1, abrogates the ability of DC to stimulate T cells in an allogeneic mixed lymphocyte reaction. Interestingly, this effect correlates with down-regulation of CD83, CD80 and CD86 surface expression during DC maturation, whereas other investigated DC surface molecules, such as MHC class I and II molecules are not significantly affected. Analysis of RNA distribution reveals that particularly the stimulated expression of CD83 depended on a functional CRM1 export receptor. Taken together, the presented data show a critical involvement of the CRM1 transport receptor in DC maturation, most likely by enabling efficient nucleo-cytoplasmic translocation of specific mRNAs. Thus, interference with this pathway may provide new strategies to modulate DC function and, subsequently, DC-mediated immune responses. PMID- 19545932 TI - Interaction of SH3P13 and DYDC1 protein: a germ cell component that regulates acrosome biogenesis during spermiogenesis. AB - The N-terminal BAR domain of endophilin has unique functions, such as affecting the curvature of the lipid membrane through its lysophosphatidic acid acyltransferase activity, binding of ATP and GTP and participating in tubulating activity. We recently demonstrated that SH3P13, a BAR domain-containing protein, assists in regulating clathrin-coated vesicle traffic that is crucial for acrosome biogenesis during spermatogenesis. DYDC1 was identified in a yeast two hybrid screen from a human testis library by using the SH3P13 BAR domain as the bait. Consistent with the expression pattern of SH3P13, DYDC1 is exclusively expressed in the brain and testis and accumulates in the acrosome area during late stage of spermiogenesis. Here, we report that DYDC1 plays a crucial role during acrosome biogenesis. This relationship has been verified by a novel approach that involves germ cell transplantation and RNA interference. We found that knockdown of endogenous Dydc1 interfered with the formation of acrosomes, and thus spermatid differentiation during mouse spermiogenesis. These data provide important insight into the crucial process of acrosome biogenesis. In addition, our approach can also be applied to study functions of other genes related to spermatogenesis in vivo. PMID- 19545933 TI - CALHM1 variant is not associated with Alzheimer's disease among Asians. AB - In a case control study involving 484 study subjects, we showed that the CALHM1 allele (13.5% vs 16.7%) and genotype frequency was not significantly different between Alzheimer's disease (AD) and controls. Logistic regression analysis did not reveal any interaction between ApoE4 allele and CALHM1 allele. PMID- 19545935 TI - Serotype replacement in perspective. AB - An increase in the incidence of invasive pneumococcal disease (IPD) caused by non vaccine serotypes (NVTs) (serotype replacement) has been observed since the introduction of the seven-valent pneumococcal conjugate vaccine (PCV7), primarily in the elderly, HIV-positive adults, and children with underlying medical conditions. In children aged <5 years, only a slight increase in replacement disease due to NVTs has been observed, but the relationship of changes in serotype carriage to replacement disease is not well understood. Predominant post PCV7 NVTs include serotypes 15, 19A, and 33F, but the increasing antibiotic resistance of 19A is of particular concern in both vaccinated and unvaccinated populations. PMID- 19545934 TI - [Structured review of physical activity measurement with questionnaires and scales in older adults and the elderly]. AB - OBJECTIVE: To characterize the distinct questionnaires and scales used to measure physical activity, their conceptual frameworks, psychometric properties and application norms. METHOD: The review included original articles that used questionnaires or scales to assess physical activity in older adults or the elderly. The CINAHL and MEDLINE databases were consulted for the years 1993 to 2007. The studies selected had to provide information on the use, development and psychometric properties of the instruments. Instruments used in the population aged more than 45 years old were included. Articles that assessed physical activity by direct estimation, complex methods, or physical performance were excluded. RESULTS: The search produced 166 references and 36 instruments were identified. Most of the studies quantified physical activity, and a minority assessed self-efficacy in physical activity performance and the stage of change. Half of the instruments were self-administered. The most frequently studied reliability was test-retest. Criterion validity was studied in 14 instruments, and 11 of these used at least two alternative methods of measurement. Construct validity was assessed in 26 instruments. Responsiveness was evaluated in only three instruments (YPAS, CHAMPS and Exercise Stage of Change) in addition to reliability and validity. CONCLUSIONS: The Exercise Stage of Change questionnaire showed responsiveness and sufficient reliability to allow individual use. The 7Day PAR questionnaire and the Modified Baecke Questionnaire (Spanish version), which evaluate physical activity, can be used on an individual basis, although their responsiveness has not been studied. In general, the instruments analyzed do not assess mild intensity activities. PMID- 19545936 TI - Correction of hemifacial microsomia with the help of mirror imaging and a rapid prototyping technique: case report. AB - A 23-year-old man presented with an 8-year history of unilateral hemifacial microsomia. A three-dimensional model of the maxillofacial bones was generated after acquisition of helical computed tomographic data. A customised implant model was designed by projecting a mirror image of the healthy mandible on to the three-dimensional model. A resin model of the implant was then made using a rapid prototyping machine. A polymeric biomaterial was sculpted according to the model and implanted into the affected side of the mandible to restore his facial symmetry. The hemifacial microsomia was corrected and a symmetrical facial contour obtained. No complications developed during the 6-year follow-up. PMID- 19545937 TI - Endoscopic removal of a supernumerary tooth from the nasal cavity in an adult. AB - Eruption of a tooth into the nasal cavity is a rare event. However it is important to identify such a condition as it has the potential to cause significant morbidity. We report a unique case of intranasal supernumerary tooth erupted into nasal causing significant nasal symptoms in an otherwise healthy patient which was removed endoscopically. PMID- 19545938 TI - Kinetics of cyanide binding as a probe of local stability/flexibility of cytochrome c. AB - Effect of anions of the Hofmeister series (thiocyanate, perchlorate, iodide, bromide, nitrate, chloride, sulfate, and phosphate) on local and global stability and flexibility of horse heart ferricytochrome c (cyt c) has been studied. Global stability of cyt c was determined by iso/thermal denaturations monitored by change in ellipticity in the far-UV region and its local stability was determined from absorbance changes in the Soret region. Particularly, relative stability/flexibility of the Met80-heme iron bond has been assessed by analysis of binding of cyanide into the heme iron. Both global and local stabilities of cyt c exhibited monotonous increase induced by a change of anion from chaotropic to kosmotropic species. However, this monotonous dependence was not observed for the rate constants of cyanide association with cyt c. As expected more chaotropic ions induced lower stability of protein and faster binding of cyanide but this correlation was reversed for kosmotropic anions. We propose that the unusual bell shaped dependence of the rate constant of cyanide association is a result of modulation of Met80-heme iron bond strength and/or flexibility of heme region by Hofmeister anions independently on global stability of cyt c. Further, our results demonstrate sensitivity of cyanide binding to local change in stability/flexibility in the heme region of cyt c. PMID- 19545940 TI - Retroperitoneoscopic transureteroureterostomy with cutaneous ureterostomy to salvage failed ileal conduit urinary diversion. AB - Reconstruction for failed urinary diversion is technically challenging, due to severe tissue adhesion around the anastomotic site. We report successful laparoscopic transureteroureterostomy with cutaneous ureterostomy via a completely extraperitoneal approach to salvage failed ileal conduit in two patients with necrotic ileal conduit and bilateral anastomotic obstruction, respectively. This novel, less invasive approach may offer a viable alternative to open surgical revision for failed ileal conduit urinary diversion. PMID- 19545939 TI - Blood-brain barrier (BBB) disruption using a diagnostic ultrasound scanner and Definity in Mice. AB - The objective of this work was to determine whether diagnostic ultrasound and contrast agent could be used to transcranially and nondestructively disrupt the blood-brain barrier (BBB) in mice under ultrasound image guidance and to quantify that disruption using magnetic resonance imaging (MRI) and magnetic resonance (MR) contrast agent. Each mouse was placed under isoflurane anesthesia and the hair on top of its skull was removed before treatment. A diagnostic ultrasound transducer was placed in a water bag coupled with gel on the mouse skull. Definity (ultrasound [US] contrast) and Magnevist (MR contrast) were injected concurrent with the start of a custom ultrasound transmission sequence. The transducer was translated along the rostral-caudal axis to insonify three spatial locations (2mm apart) along one half of the brain for each sequence. T1-weighted MR images were used to quantify the volume of tissue over which the BBB disruption allowed Magnevist to enter the brain, based upon increases in MR contrast-to-noise ratio (CNR) compared with the noninsonified portions of the brain. Ultrasonic frequency, pressure and pulse duration, as well as Definity dose and injection time were varied. Preliminary results suggest that a threshold exists for BBB opening dependent upon both pressure and pulse duration (consistent with reports in the literature performed at lower frequencies). A range of typical diagnostic frequencies (e.g., 5.0-8.0 MHz) generated BBB disruption. Comparable BBB opening was noted with varied delays between Definity injection and insonification (0-2 min) for a range of Definity concentrations (400-2400 microL/kg). The low-pressure, custom sequences (mechanical index [MI]< or =0.65) had minimal blood cell extravasation as determined by histologic evaluation. This study has shown the ability of a diagnostic ultrasound system, in conjunction with Definity, to open the BBB transcranially in a mouse model for molecules approximately 0.5 kDa in size. Opening was achieved at higher frequencies than previously reported and was localized under ultrasound image guidance. A typical, ultrasound imaging mode (pulsed wave [PW] Doppler) with specific settings (transmit frequency=5.7 MHz, gate size=15 mm, pulse repetition frequency=100 Hz, system power=15%) successfully opened the BBB, which facilitates implementation using the most of commercially available clinical diagnostic scanners. Localized opening of the BBB may have potential clinical utility for the delivery of diagnostic or therapeutic agents to the brain. PMID- 19545941 TI - Surveillance in stage I seminoma patients: a long-term assessment. AB - BACKGROUND: Following orchidectomy patients with stage I seminoma of the testis may be managed by either surveillance or adjuvant treatment. In view of the very high cure rate, it is important to analyse long-term outcomes. OBJECTIVE: To provide data to advise patients on treatment burden and risk of recurrence associated with surveillance. DESIGN, SETTING, AND PARTICIPANTS: We audited the case records of 164 stage I seminoma patients registered at the Royal Marsden Hospital who were managed with a surveillance policy between 1980 and 2004 and followed for 1-20 yr (median: 13.5 yr). MEASUREMENTS: All treatments and patterns of relapse were documented. RESULTS AND LIMITATIONS: Twenty-two of 164 (13%) patients had relapsed at a median of 15.5 mo (range: 6-55 mo) from orchidectomy. Eighteen relapses appeared to be confined to the para-aortic nodes, but 6 of the 13 (46%) men treated with only para-aortic radiotherapy suffered a further relapse at another site. The disease-specific mortality was 1.3%. In the complete series of 164 patients, a total of 50 cycles of chemotherapy and 26 courses of radiotherapy was administered, representing an average of 0.46 "treatment units" per patient or an average of 3.45 treatment units per relapsing patient. The total number of treatment days was 390 d for radiotherapy and 133 d for chemotherapy, representing an average of 3.2 d per patient or 23.8 d per relapsing patient. This was a single-centre series extending back to the 1980s. Imaging and treatment protocols have advanced since then. CONCLUSIONS: Surveillance postorchidectomy is a safe practice in the long term, and the majority of patients can avoid further treatment. There is the risk that those who do relapse face a higher burden of treatment than would be required if adjuvant treatment had been given. PMID- 19545942 TI - Reductive dechlorination of hexachlorobenzene by Cu/Fe bimetal in the presence of nonionic surfactant. AB - This study investigated the enhancement of nonionic surfactant (TritonX-100, TX 100) on the reductive dechlorination of hexachlorobenzene (HCB) by microscale Cu/Fe bimetal. HCB reduction by Cu/Fe without TX-100, in aqueous TX-100, adsorbed with TX-100 and prepared with TX-100 was compared. Results showed that HCB reduction by Cu/Fe in aqueous TX-100 increased with increasing TX-100 concentration, reached a maximum of 98.6% when aqueous TX-100 concentration attained its critical micelle concentration (CMC), and decreased with the further increase in TX-100 concentration. The mechanism was suggested as the increased HCB mass transport at low TX-100 concentration below CMC and the coverage of reactive sites at high TX-100 concentration above CMC. The greatest reduction was achieved by Cu/Fe in the presence of 0.2mM TX-100, followed by Cu/Fe adsorbed and prepared with 0.2 mM TX-100. Pentachlorobenzene and tetrachlorobenzene were found as the main dechlorination intermediates. Successive reduction by Cu/Fe in aqueous TX-100 solution could be maintained above 98% for four cycles. It is an alternative to use nonionic surfactants to enhance the reductive dechlorination of hydrophobic polychlorinated aromatic hydrocarbons (PCAH) by Cu/Fe bimetal. PMID- 19545943 TI - Effects of reduction products of ortho-hydroxyl substituted azo dyes on biodecolorization of azo dyes. AB - The mediated effects of reduction products of some ortho-hydroxyl substituted azo dyes on biodecolorization were investigated. The results indicated that the addition of reduction products could effectively accelerate dye decolorization by Shigella sp. QRZ-1. The best accelerating effect was obtained with the addition of reduction products of Acid Red 14 (AR14), resulting in an over 3-fold increase in decolorization efficiency of many azo dyes. In sequencing batch reactor experiments, the accelerating effect of reduction products of AR14 was more obvious (1.5-fold) during the startup of the system. When the dye concentration was increased to 500 mg L(-1), the accelerated decolorization efficiency was still maintained around 95%. The presence of AR14 in the feed enhanced the decolorization performance of anaerobic sludge, indicating that the strategy may be beneficial for practical application. 1-Naphthol-2-amino-4-sulfonic acid, which is one of the reduction products of AR14, may function as redox mediator to speed up azo dye biodecolorization. PMID- 19545944 TI - Nitrate contamination in groundwater of some rural areas of Rajasthan, India. AB - Efforts were made to evaluate the level of nitrate in some agro-economy based rural habitations of northern Rajasthan, India. A total of 64 groundwater samples from 21 different villages/sub-villages of district Sri Ganganagar, India were collected and analyzed for nitrate (as NO(3)(-)), sulphate (as SO(4)(2-)) and few other parameters. NO(3)(-) level in groundwater was 7.10-82.0 mg l(-1) for individual samples. But average NO(3)(-) for total samples was 60.6+/-33.6 (SD) mg l(-1), which indicates the non-suitability of groundwater for drinking purposes, if BIS permissible limit (22.6 mg l(-1)) is considered as reference level. SO(4)(2-) ranged form 28.6 to 660.3 mg l(-1) in this area. The regression analysis indicates the difference sources for NO(3)(-) and SO(4)(2-) contamination in different regions rather than a common source. The point and non point sources of NO(3)(-) and SO(4)(2-) in groundwater of this region may be N fertilizer, sewerage, animal waste, organic manure, geology of sub-surface soil layers, pit latrines, etc. Results thus indicated that groundwater of this part of the State is severely polluted due to anthropogenic activities. The continuous consumption of such water may pose serious health hazardous in local residents. PMID- 19545946 TI - In vitro culture combined with quantitative TaqMan PCR for the assessment of Toxoplasma gondii tissue cyst viability. AB - Toxoplasma gondii is a serious food-borne pathogen with a worldwide distribution. In order to assess the risk of contracting toxoplasmosis from certain foods, many studies rely on the molecular detection of T. gondii DNA. However, determining the viability of parasites in positive samples is much more problematic. In this paper we describe a novel viability assay that relies on semi-quantitative comparison of the amount of parasite DNA present in samples used to infect host cell monolayers in vitro, and the amount of DNA detected in the same monolayers after 23 days incubation. Our assay is robust, easy to perform and interpret and offers a viable alternative to bioassays, for use in epidemiological studies, or the evaluation of specific food safety treatments. PMID- 19545945 TI - Electrochemical reclamation of silver from silver-plating wastewater using static cylinder electrodes and a pulsed electric field. AB - Silver was reclaimed from silver-plating wastewater by using a pulsed electric field (PEF) combined with static cylinder electrodes (SCE). The conditions that produced the maximal silver recovery rate (RR(Ag)) (99%) were as follows: average retention time of 10 min, interelectrode gap of 50mm, solution pH of 9.0, temperature of 45 degrees C, initial Ag(I) concentration of 1000 mg L(-1), PEF pulse frequency of 1200 Hz, current density of 5.0 A m(-2) and a pulse duty cycle of 60%. Compared with the conventional direct current (DC) technology, the PEF process exhibited improvements in the silver recovery rate (RR(Ag)), total energy consumption (TEC) and physical properties of the silver deposits, especially for low Ag(I) concentrations, for example, from 500 to 1000 mg L(-1). For an initial Ag(I) concentration of 500 mg L(-1), the PEF process produced an RR(Ag) of up to 99%, and the TEC was 4.56 kWh (kg Ag)(-1). In comparison, the RR(Ag) and TEC were 90% and 5.66 kWh (kg Ag)(-1), respectively, in the DC process. The results of SEM observation and XRD analysis indicated that the silver deposits formed by the PEF process were smaller, denser, and of a higher purity than those produced by the DC process. Therefore, the presented method was effective for reclaiming silver from silver-plating wastewater. PMID- 19545948 TI - Vishweshwaraiah Iron Steel Limited (VISL) fire disasters following ladle refining furnace accident, July 12, 1996. PMID- 19545949 TI - Methodological quality of randomised controlled trials in burns care. A systematic review. AB - OBJECTIVE: To evaluate the methodological quality of published randomised controlled trials (RCTs) in burn care treatment and management. METHODS: Using a predetermined search strategy we searched Ovid MEDLINE (1950 to January 2008) database to identify all English RCTs related to burn care. Full text studies identified were reviewed for key demographic and methodological characteristics. Methodological trial quality was assessed using the Jadad scale. RESULTS: A total of 257 studies involving 14,535 patients met the inclusion criteria. The median Jadad score was 2 (out of a best possible score of 5). Information was given in the introduction and discussion sections of most RCTs, although insufficient detail was provided on randomisation, allocation concealment, and blinding. The number of RCTs increased between 1950 and 2008 (Spearman's rho=0.6129, P<0.001), although the reporting quality did not improve over the same time period (P=0.1896) and was better in RCTs with larger sample sizes (median Jadad score, 4 vs. 2 points, P<0.0001). Methodological quality did not correlate with journal impact factor (P=0.2371). CONCLUSIONS: The reporting standards of RCTs are highly variable and less than optimal in most cases. The advent of evidence-based medicine heralds a new approach to burns care and systematic steps are needed to improve the quality of RCTs in this field. Identifying and reviewing the existing number of RCTs not only highlights the need for burn clinicians to conduct more trials, but may also encourage burn health clinicians to consider the importance of conducting trials that follow appropriate, evidence-based standards. PMID- 19545951 TI - Vitamin D, a neuro-immunomodulator: implications for neurodegenerative and autoimmune diseases. AB - It has been known for more than 20 years that vitamin D exerts marked effects on immune and neural cells. These non-classical actions of vitamin D have recently gained a renewed attention since it has been shown that diminished levels of vitamin D induce immune-mediated symptoms in animal models of autoimmune diseases and is a risk factor for various brain diseases. For example, it has been demonstrated that vitamin D (i) modulates the production of several neurotrophins, (ii) up-regulates Interleukin-4 and (iii) inhibits the differentiation and survival of dendritic cells, resulting in impaired allo reactive T cell activation. Not surprisingly, vitamin D has been found to be a strong candidate risk-modifying factor for Multiple Sclerosis (MS), the most prevalent neurological and inflammatory disease in the young adult population. Vitamin D is a seco-steroid hormone, produced photochemically in the animal epidermis. The action of ultraviolet light (UVB) on 7-dehydrocholesterol results in the production of pre-vitamin D which, after thermo-conversion and two separate hydroxylations, gives rise to the active 1,25-dihydroxyvitamin D. Vitamin D acts through two types of receptors: (i) the vitamin D receptor (VDR), a member of the steroid/thyroid hormone superfamily of transcription factors, and (ii) the MARRS (membrane associated, rapid response steroid binding) receptor, also known as Erp57/Grp58. In this article, we review some of the mechanisms that may underlie the role of vitamin D in various brain diseases. We then assess how vitamin D imbalance may lay the foundation for a range of adult disorders, including brain pathologies (Parkinson's disease, epilepsy, depression) and immune-mediated disorders (rheumatoid arthritis, type I diabetes mellitus, systemic lupus erythematosus or inflammatory bowel diseases). Multidisciplinary scientific collaborations are now required to fully appreciate the complex role of vitamin D in mammal metabolism. PMID- 19545952 TI - Thyroid stimulating hormone and prospective memory functioning in old age. AB - Alterations of thyroid functioning are common in old age. Even among persons free from thyroid disorders, subclinical variations in thyroid functioning may affect cognitive performance. However, it is unknown whether prospective memory (ProM) is related to thyroid stimulating hormone (TSH) variations. An association could be expected, as changes in the thyroid gland have been linked to alterations in frontal brain regions that play a key role in prospective remembering. Thus, the aim of this study was to examine whether subclinical variations in thyroid functioning affect ProM performance. We studied 103 participants, 75 years and older, who were free from thyroid disorders and had serum levels of TSH and thyroxine (T4) within normal ranges. Interestingly, we found a non-linear association between TSH and ProM performance, where persons with TSH levels above the fourth quartile performed substantially better than persons in the other quartiles. T4 levels were unrelated to ProM performance. This pattern suggests that the previously identified "normal-range" interval for TSH should be moved further up in old age, at least when cognitive functioning is considered. PMID- 19545953 TI - Same ablation may be used for frequent premature ventricular contractions and supraventricular tachycardia. AB - Radiofrequency catheter ablation (RFCA) has been increasingly used for the treatment of various kinds of cardiac arrhythmias over the past 20 years. RFCA of symptomatic premature ventricular contractions (PVCs) has been reported to be a safe and effective treatment option and may be considered in patients with extremely symptomatic or incapacitating cases who do not respond to medications. Long-term prognosis in patients with truly idiopathic PVCs is excellent, despite frequent recurrences of tachycardia. Sudden death is rare in patients with initially normal left and right ventricular function, in such patients, occult cardiomyopathy is usually identified on postmortem examination. Similarly, progression to diffuse cardiomyopathy is rare. Ablation of PVCs may equal that of supraventricular tachycardia in terms of success rate, safety. These new views will help understand the diagnosis and treatment method for frequent PVCs in patients without heart disease. PMID- 19545950 TI - Corepressors, nuclear receptors, and epigenetic factors on DNA: a tail of repression. AB - The differential exposure to circulating steroid hormones during brain development can have lasting consequences on brain function and behavior; therefore, the tight control of steroid hormone action within the developing brain is necessary for the expression of appropriate sex-typical behavior patterns later in life. The restricted control of steroid hormone action at the level of the DNA can be accomplished through the recruitment of coregulatory complexes. Nuclear receptor action can either be enhanced by the recruitment of coactivator complexes or suppressed by the formation of corepressor complexes. Alternatively, the regulation of nuclear receptor-mediated gene transcription in the developing brain may involve a dynamic process of coactivator and corepressor function on DNA. It is likely that understanding how different combinations of coregulatory matrixes assembly on DNA will lead to further understanding of heterogeneous responses to nuclear receptor activation. We will discuss how coregulators influence gene transcription and repression, the role of chromatin binding factors in the regulation of gene transcription, and their potential impact on brain development. PMID- 19545954 TI - The most efficient follicular regenerating unit and the smallest follicular regenerating unit: potential treatments for hair loss. AB - Hair loss affects many people, especially adult males. An effective treatment is hair transplantation which involves harvesting hair grafts from a donor site and relocating them to a bald site. However, this traditional method, equivalent to one-to-one transplantation, simply redistributes hair rather than increases the number of existing hairs. Although hair transplantation is actually the transplantation of hair follicle (HF), it has been confirmed that whole HFs could be reformed from parts of HFs containing different constituents, implying the existence of more efficient and smaller HF regenerating units in a whole HF. Thus we hypothesize that the most efficient follicular regenerating unit (EFRU) and the smallest follicular regenerating unit (SFRU) could be found in whole HFs. As a result, the one-to-many hair transplantation would be achieved in clinic. One to-many means to double or triple the number of hairs. In order to test and verify the hypothesis, we design a method called hair follicle micro-dissection (HFM) which aims to help find the regenerating units and increase the number of hair for transplantation. The postulation may provide a more mature and realistic treatment for hair loss if it proved to be practical. PMID- 19545955 TI - Are female sex steroids neuroprotective in experimental stroke. AB - The central nervous system and the peripheral nervous system are important targets of gonadal steroids, which affect brain development and differentiation, and influence neuronal functions. Most of the studies have shown that female sex steroids progesterone and 17beta-estradiol exert protective effects in the experimental models of stroke, although deleterious effects have also been reported. Extensive studies on female sex steroids in cerebral ischemia have described their potential to act on multiple ischemic injury pathways, and pointed out the significance of numerous factors, including gender, age, brain region, duration of ischemia, and precise dose of steroids. PMID- 19545956 TI - Comment on "Alzheimer's disease and atherosclerosis: passers-by or brothers?". PMID- 19545957 TI - Abdominal angiostrongyliasis caused by Angiostrongylus cantonensis: a possible cause of eosinophilic infiltration in human digestive tract. AB - We report the pathological findings of a serologically proven case of Angiostrongylus cantonensis presenting with localized peritonitis followed by eosinophilic meningoencephalitis. The neurological involvement developed 3 days after the occurrence of gastrointestinal symptoms. Similarly to the life cycle in rats, it takes about 2 or 3 days for the larvae to reach the nervous system. The pathological section of the sigmoid colon showed focal eosinophilic infiltration with serosal vessel invasion. In the case reported here, we describe a new possible cause of eosinophilic infiltration in the human digestive tract. PMID- 19545958 TI - Assess the discrimination of Achilles InSight calcaneus quantitative ultrasound device for osteoporosis in Chinese women: compared with dual energy X-ray absorptiometry measurements. AB - Since the implementation of quantitative ultrasound (QUS) technology may become a part of future clinical decision making to identify osteoporosis and prevent fractures, this study was initiated to evaluate the correlations of QUS parameters and axial bone mineral density (BMD) using dual energy X-ray absorptiometry (DXA) and to assess the discrimination of QUS measurements for osteoporosis and osteopenia defined by WHO criteria. 106 native Chinese women (aged 50.2+/-10.9 SD, 21-74 years) were involved. Each subject received both QUS measurements at left calcaneus with Achilles InSight and DXA measurements with DPX-L at lumbar spine (L(2-4)), total hip and femoral neck. Achilles InSight provided the stiffness index (SI) which derived from Broadband Ultrasound Attenuation (BUA) and Speed of Sound (SOS), and the T-scores of SI were calculated. We found that the QUS parameter SI was statistically significant but medium correlated (r=0.458-0.587) with DXA at the lumbar spine, total hip and femoral neck (P<0.0001 for all correlations). With ROC analysis, the area under the ROC curve of diagnosis of osteoporosis and osteopenia were 0.933 and 0.796, respectively. To identify osteoporosis, when the T-score threshold of SI was defined as -1.4, the sensitivity was 100%, and the specificity was 73.7%. Our study confirmed that QUS measurements performed with Achilles InSight were capable to identify osteoporosis defined by axial BMD using DXA in Chinese women. PMID- 19545959 TI - Mechanic effect of pulsed focused ultrasound in tumor and muscle tissue evaluated by MRI, histology, and microarray analysis. AB - The purpose of this study was to investigate the effect of pulsed high-intensity focused ultrasound (HIFU) to tumor and muscle tissue. Pulsed HIFU was applied to tumor and muscle tissue in C3H/Km mice. Three hours after HIFU treatment pre- and post-contrast T1-wt, T2-wt images and a diffusion-wt STEAM-sequence were obtained. After MR imaging, the animals were euthenized and the treated tumor and muscle was taken out for histology and functional genomic analysis. In the tumor tissue a slight increase of the diffusion coefficient could be found. In the muscle tissue T2 images showed increased signal intensity and post-contrast T1 showed a decreased contrast uptake in the center and a severe contrast uptake in the surrounding muscle tissue. A significant increase of the diffusion coefficient was found. Gene expression analysis revealed profound changes in the expression levels of 29 genes being up-regulated and 3 genes being down-regulated in the muscle tissue and 31 genes being up-regulated and 15 genes being down regulated in the SCCVII tumor tissue. Seven genes were up-regulated in both tissue types. The highest up-regulated gene in the tumor and muscle tissue encoded for Mouse histone H2A.1 gene (FC=13.2+/-20.6) and Apolipoprotein E (FC=12.8+/-27.4) respectively MHC class III (FC=83.7+/-67.4) and hsp70 (FC=75.3+/ 85.0). Immunoblot confirmed the presence of HSP70 protein in the muscle tissue. Pulsed HIFU treatment on tumor and muscle tissue results in dramatic changes in gene expression, indicating that the effect of pulsed HIFU is in some regard dependent and also independent of the tissue type. PMID- 19545960 TI - MR appearance of normal and abnormal bile: correlation with imaging and endoscopic finding. AB - Identification of abnormal bile related to various pathological processes in the pancreaticobiliary tract can be important in the diagnosis of disease and the determination of appropriate treatment. Magnetic resonance (MR) imaging can allow comprehensive evaluation of abnormal bile because MR usually provides better tissue characterization than other imaging modalities. A high-intensity signal from bile is frequently encountered on T1-weighted images and can be seen in concentrated bile, sludge, stones, or hemobilia. Contrast-enhanced MR features, such as inhomogeneous hepatic enhancement in the arterial phase and papillitis or mild-to-moderate bile duct wall thickening with enhancement, along with clinical characteristics, may suggest clinically significant bile, such as sludge or purulent bile, rather than merely concentrated bile. A history of trauma and appropriate imaging findings in the hepatobiliary tract can support a diagnosis of hemobilia. MR imaging may assist in diagnosing intraductal papillary mucinous neoplasm of the bile duct via detection of an intraductal mass or several indirect signs, suggesting a large amount of mucin. Additionally, Gd-EOB-DTPA enhanced MR may delineate mucin as a filling defect surrounding hyperintense bile. A floating filling defect on all MR sequences is helpful in discriminating pneumobilia from other intraluminal filling defects. Familiarity with the various different MR features of abnormal bile signals can therefore facilitate accurate diagnosis and treatment. PMID- 19545961 TI - Novel inclusion complex of ibuprofen tromethamine with cyclodextrins: physico chemical characterization. AB - Guest-host interactions of ibuprofen tromethamine salt (Ibu.T) with native and modified cyclodextrins (CyDs) have been investigated using several techniques, namely phase solubility diagrams (PSDs), proton nuclear magnetic resonance ((1)H NMR), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), X-ray powder diffractometry (XRPD), scanning-electron microscopy (SEM) and molecular mechanics (MM). From the analysis of PSD data (A(L)-type) it is concluded that the anionic tromethamine salt of ibuprofen (pK(a)=4.55) forms 1:1 soluble complexes with all CyDs investigated in buffered water at pH 7.0, while the neutral form of Ibu forms an insoluble complex with beta-CyD (B(S)-type) in buffered water at pH 2.0. Ibu.T has a lower tendency to complex with beta-CyD (K(11)=58 M(-1) at pH 7.0) compared with the neutral Ibu (K(11)=4200 M(-1)) in water. Complex formation of Ibu.T with beta-CyD (DeltaG(o)= 20.4 kJ/mol) is enthalpy driven (DeltaH(o)=-22.9 kJ/mol) and is accompanied by a small unfavorable entropy (DeltaS(o)=-8.4 J/mol K) change. (1)H NMR studies and MM computations revealed that, on complexation, the hydrophobic central benzene ring of Ibu.T and part of the isobutyl group reside within the beta-CyD cavity leaving the peripheral groups (carboxylate, tromethamine and methyl groups) located near the hydroxyl group networks at either rim of beta-CyD. PSD, (1)H NMR, DSC, FT-IR, XRPD, SEM and MM studies confirmed the formation of Ibu.T/beta CyD inclusion complex in solution and the solid state. PMID- 19545962 TI - Development and validation of a stability-indicating HPLC method for simultaneous determination of salicylic acid, betamethasone dipropionate and their related compounds in Diprosalic Lotion. AB - Diprosalic Lotion is an anti-inflammatory drug product that contains salicylic acid and betamethasone dipropionate as active pharmaceutical ingredients (APIs). A reversed-phase high performance liquid chromatography (RP-HPLC) method was developed for simultaneous determination of salicylic acid, betamethasone dipropionate, and their related compounds in Diprosalic Lotion. A 150 mm x 4.6 mm I.D. YMC J'sphere ODS-H80 column at 35 degrees C and UV detection at 240 nm was used. A gradient elution was employed using 0.05% (v/v) methanesulfonic acid solution and acetonitrile as mobile phases. A total of thirty three compounds from Diprosalic Lotion samples were separated in 38 min. The stability-indicating capability of this method has been demonstrated by the adequate separation of all the impurities and degradation products in expired stability samples of Diprosalic Lotion. The method was validated as per the current ICH guidelines. PMID- 19545964 TI - Re: "Are one or two dangerous? Diphenoxylate-atropine exposure in toddlers". PMID- 19545963 TI - Optimization of a method for determination of phenolic acids in exotic fruits by capillary electrophoresis. AB - In this work, the separation of nine phenolic acids (benzoic, caffeic, chlorogenic, p-coumaric, ferulic, gallic, protocatechuic, syringic, and vanillic acid) was approached by a 3(2) factorial design in electrolytes consisting of sodium tetraborate buffer (STB) in the concentration range of 10-50 mmol L(-1) and methanol in the volume percentage of 5-20%. Derringer's desirability functions combined globally were tested as response functions. An optimal electrolyte composed by 50 mmol L(-1) tetraborate buffer at pH 9.2, and 7.5% (v/v) methanol allowed baseline resolution of all phenolic acids under investigation in less than 15 min. In order to promote sample clean up, to preconcentrate the phenolic fraction and to release esterified phenolic acids from the fruit matrix, elaborate liquid-liquid extraction procedures followed by alkaline hydrolysis were performed. The proposed methodology was fully validated (linearity from 10.0 to 100 microg mL(-1), R(2)>0.999; LOD and LOQ from 1.32 to 3.80 microg mL(-1) and from 4.01 to 11.5 microg mL(-1), respectively; intra-day precision better than 2.8% CV for migration time and 5.4% CV for peak area; inter day precision better than 4.8% CV for migration time and 4.8-11% CV for peak area; recoveries from 81% to 115%) and applied successfully to the evaluation of phenolic contents of abiu-roxo (Chrysophyllum caimito), wild mulberry growing in Brazil (Morus nigra L.) and tree tomato (Cyphomandra betacea). Values in the range of 1.50-47.3 microg g(-1) were found, with smaller amounts occurring as free phenolic acids. PMID- 19545965 TI - Thyroid gland rupture: a rare case of respiratory distress. AB - BACKGROUND: Rupture of a normal thyroid gland after blunt cervical trauma is a rare case of airway compression. This case report describes the case of a 79-year old woman who developed severe respiratory distress after a bicycle crash. CASE REPORT: Presenting about 2 h after the crash, the patient noted cervical swelling and increasing dyspnea. The diagnosis of thyroid gland rupture was made with a combination of computed tomography scan, cervical radiography, and bronchoscopy. Invasive airway management was required in the first few hours after the crash. The patient underwent a left hemithyroidectomy, and recovered without complications. CONCLUSION: This case report highlights the fact that thyroid gland rupture can be a threatening complication of blunt cervical trauma. PMID- 19545966 TI - Intraosseous access. AB - BACKGROUND: Vascular access is of paramount importance in the care of the critically ill patient. When central or peripheral intravenous access cannot be accomplished in a timely manner, intraosseous access and infusion is a rapid and safe alternative for the delivery of fluids, medications, and blood products. The resurgence of the use of intraosseous access in the 1980s led to the development of new methods and devices that facilitate insertion. OBJECTIVES: This article discusses general indications, contraindications, and complications of intraosseous access and infusion, focusing on new devices and their insertion. DISCUSSION: Current research is focused on product innovation and improving drug delivery using intraosseous autoinjectors, finding new anatomic sites for placement, and expanding the use of different intraosseous devices to the adult population. CONCLUSIONS/SUMMARY: New, improved intraosseous systems provide health care providers with choices beyond traditional manual intraosseous access for administering fluids. PMID- 19545967 TI - Does early intensive lowering of blood pressure reduce hematoma volume and improve clinical outcome after acute cerebral hemorrhage? AB - BACKGROUND: Intracerebral hemorrhage (ICH) is a poorly understood condition with devastating results. Despite the personal and social impact of ICH, modern medicine can offer little hope. Surgery is the longest-standing therapy, but with no demonstrated evidence of positive effect. Reduction of the early hypertension seen with ICH is believed to limit hematoma growth and improve clinical outcome. The effectiveness and safety of an early, aggressive blood-pressure-lowering strategy for ICH patients has only recently been examined in randomized controlled trials. CLINICAL QUESTION: Does early, intensive lowering of blood pressure reduce hematoma volume and improve clinical outcome after acute cerebral hemorrhage? EVIDENCE REVIEW: Two trials assessing the clinical impact of blood pressure lowering in intracerebral hemorrhage were acquired and appraised. RESULTS: The two randomized trials incorporated similar parallel designs and both trials measured clinical outcomes and short-term change in hematoma size. The smaller trial was only partially blinded and showed no difference in any of the outcomes; the large trial demonstrated marginal reduction in proportional hematoma growth, but no significant difference in clinical outcome. CONCLUSIONS: There is currently insufficient evidence to support the routine practice of lowering blood pressure of patients suffering an acute intracerebral hemorrhage. PMID- 19545968 TI - A 3-month-old female with an inguinal mass. AB - BACKGROUND: Inguinal masses are a common finding among infants. The differential diagnosis of these masses in infants is broad, with inguinal hernia being the most common diagnosis in both males and females. However, the evaluation and management of males vs. females with inguinal masses is somewhat different due to the greater potential for gonad involvement in males. OBJECTIVES: The pathophysiology and management of inguinal hernias is discussed with a specific focus on inguinal hernias in females. CASE REPORT: We present a case of a 3-month old girl with an inguinal hernia and a mass, found to be an incarcerated ovary. CONCLUSIONS: Inguinal masses in infancy are common, with inguinal hernia being the most common cause by far. A female infant with suspected inguinal hernia should be thoroughly evaluated to determine whether ovarian content is present. PMID- 19545969 TI - Right atrial and inferior vena caval thrombosis in a case of amebic liver abscess. PMID- 19545970 TI - A comparison of HIV testing in the emergency department and urgent care. AB - BACKGROUND: Although national guidelines recommend universal human immunodeficiency virus (HIV) testing, emergency departments (EDs) may choose to limit testing to certain patients, such as those triaged to urgent care (UC). OBJECTIVE: To compare the results of rapid HIV testing in an urban ED with an affiliated UC. METHODS: This was a retrospective analysis of an HIV testing program that included screening, which was initiated by triage nurses, and diagnostic testing, which was initiated by clinicians. Eligible patients were >= 12 years old and medically stable. RESULTS: From April 2005 through December 2006, HIV tests were completed in 6196 (8.3%) of the 74,331 ED visits and 3256 (8.8%) of the 37,169 UC visits. Screening accounted for 5009 (80.8%) of the ED tests and 2914 (89.5%) of the UC tests, and diagnostic testing accounted for the remainder. Eighty (1.3%) of the ED tests and 21 (0.6%) of the UC tests were positive (p = 0.0024). Compared with newly diagnosed HIV-positive ED patients, HIV-positive UC patients were less likely to have CD4 counts < 200 cells/MUL (adjusted odds ratio 0.19, 95% confidence interval 0.05-0.65). CONCLUSION: Although the yield of HIV testing is greater among ED patients, UC patients are diagnosed at a less advanced stage of illness. PMID- 19545971 TI - Palytoxin poisoning after dermal contact with zoanthid coral. AB - BACKGROUND: Palytoxin is most commonly reported after ingestion of seafood. We report a case of palytoxin poisoning from dermal absorption with local toxicity from zoanthid coral in a patient with intact skin. CASE REPORT: A 25-year-old previously healthy woman handled a zoanthid coral from a home aquarium without any barrier protection. The patient manifested neurologic symptoms of perioral paresthesia and dysguesia. In addition, there was local dermatologic toxicity that persisted for several days. The patient was treated supportively with corticosteroids and a histamine antagonist. CONCLUSION: We report a case of palytoxin poisoning from dermal absorption after handling a zoanthid coral. Palytoxin is a potent marine toxin that affects the sodium-potassium ATPase (adenosinetriphosphatase) pump and can cause multiple clinical effects, including paresthesia, dysguesia, hypertension, respiratory depression, coma, and death. PMID- 19545973 TI - Observer-rated rapport in interactions between medical students and standardized patients. AB - OBJECTIVE: To measure rapport between medical students and standardized patients using observer ratings; to relate these ratings to students' emotional awareness and to behavior within the medical interaction; and to assess the relative validity of using excerpts of different lengths for the measurement of rapport. METHODS: Third-year medical students (N=141) were videotaped during a 15-min interaction with a standardized patient, and rapport as well as other communication variables were measured using trained coders. Rapport was measured with good interrater reliability by trained coders who viewed three 1-min excerpts. Emotional awareness was measured by a test of recognizing facial expressions of emotion, self-ratings of emotional self-awareness, and peer ratings of interpersonal sensitivity. Finally, participants who viewed the videotapes while imagining themselves to be the patient (analogue patients) provided impressions of the students, including satisfaction. RESULTS: Rapport based on all three minutes was positively correlated with accuracy in decoding facial expressions of emotion, self-reported attention to one's own emotions, peer ratings of sensitivity, communication behaviors of the medical student and standardized patient, and analogue patients' positive impressions and satisfaction. Rapport based on just the first minute of the interaction was significantly related to many of these variables. CONCLUSION: Rating short excerpts of behavior is a valid and efficient methodology for capturing the concept of rapport. PRACTICE IMPLICATIONS: Clinicians, researchers, and educators should focus on rapport building, even very early in the medical visit. PMID- 19545972 TI - Prevalence and correlates of mothers and fathers attending pretest cancer genetic counseling together. AB - OBJECTIVE: To determine the prevalence of fathers' attendance at pretest cancer genetic counseling sessions with mothers undergoing BRCA1/2 genetic testing for hereditary breast/ovarian cancer (HBOC) risk, and to identify psychosocial and other correlates of fathers' attendance. METHODS: One hundred and twenty-one fathers of minor-age children who were spouses/partners of women (mothers) undergoing such counseling and testing were recruited, completed a behavioral self-report survey, and provided data about their sociodemographic backgrounds, father-child cancer communication histories, parenting relationship quality, and information-seeking and perceived knowledge. RESULTS: A total of 27.3% of fathers attended pretest cancer genetic counseling with mothers. Compared to fathers who did not attend pretest cancer genetic counseling, those who did had stronger parenting alliances with mothers, were more likely to have sought out information about BRCA1/2 testing, and felt more informed about testing. In an adjusted logistic regression model of session attendance, the strength of the parenting alliance was associated with a 6% increase in the likelihood of attending genetic counseling (odds ratio [OR]=1.06, 95% confidence interval [CI]=1.01, 1.12, p<.05) and greater perceived knowledge about BRCA1/2 testing was associated with a four fold increase in the likelihood of session attendance (OR=4.03, CI=1.77, 9.37, p<.001). CONCLUSION: One in three fathers attend pretest cancer genetic counseling with mothers undergoing BRCA1/2 testing; those who do have closer parenting relationships and are more informed about BRCA1/2 testing. PRACTICE IMPLICATIONS: When possible, providers should discuss mothers including fathers in cancer genetic counseling sessions as this may affect outcomes of HBOC genetic counseling and testing. PMID- 19545975 TI - Quantification of enhancement of left ventricular myocardium in patients with dilated cardiomyopathy using delayed enhanced MR imaging. AB - The purpose is to evaluate delayed enhancement (DE) of the myocardium in patients with dilated cardiomyopathy (DCM), compared with control subjects. We also evaluated the interrelationships of DE and contractile function. DCM patients (n = 42) and 14 control subjects were evaluated by DE MR imaging, acquired using a two-dimensional segmented inversion-recovery prepared gradient-echo sequence (TI = 250 ms), 15 min after intravenous administration of 0.2 mmol/kg gadolinium. For the myocardium of left ventricle (LV), we traced epicardial and endocardial borders, and regions of interest (ROIs) were placed in each slice. For analysis of DE images, the signal-to-noise ratio (SNR) and the contrast-to-noise ratio (CNR) of the LV myocardium were calculated. The averaged SNR (aSNR) and averaged CNR (aCNR) per slice of the LV myocardium were calculated. In the DCM group, we also evaluated the interrelationship of DE and the contractile function of the LV. Mean aSNR was not significantly different between the studied groups; however, mean aCNR was significantly higher in the DCM group (3.5+/-3.1) than in control subjects (-4.1+/-2.1). In the DCM group, aCNR was moderately related to LV ejection fraction (LVEF) (r = 0.52, P<.0001). Mean aCNR was significantly higher in the DCM group with low LVEF (<25%) (6.0+/-2.8) than in the DCM group with high LVEF (>or=25%) (2.0+/-2.3). In DE MR imaging, the LV myocardium of DCM usually has high aCNR, which may suggest fibrosis. Quantification of aCNR may contribute to the diagnosis of DCM. The level of aCNR seems to correlate with LVEF. Using this technique, quantification of aCNR is objective and very useful for the diagnosis of DCM and contractile function of LV. PMID- 19545974 TI - Changes in surrogate outcomes can be translated into clinical outcomes using a Monte Carlo model. AB - OBJECTIVE: To meaningfully interpret trials using surrogate outcomes, one must translate changes in the surrogate outcome to changes in the clinical outcome. Formulae to do this are uncommon because they require primary data from multiple randomized trials that measure both the surrogate and clinical outcome. STUDY DESIGN AND SETTING: We developed a model to translate changes in anticoagulation control (the surrogate outcome) into hemorrhagic and thromboembolic event rates (the clinical outcome). The model used Monte Carlo simulation and association measures between the surrogate and the clinical outcome from a meta-analysis. In randomized trials having interventions that improved anticoagulation control, we used the model to predict and statistically compare event rates between the study groups. RESULTS: Seven randomized trials found significantly improved anticoagulation control (mean increase in proportion of time in therapeutic range: 8.4%; range: 1.8-18%). These improvements in anticoagulation control translated to small decreases in hemorrhagic and thromboembolic events (mean: 0.66%/yr; range: 0.13-1.42%). These changes were never statistically significant. CONCLUSION: Monte Carlo modeling can be used to translate surrogate outcomes into clinical outcomes. Statistically significant changes in anticoagulation control did not translate to significant differences in clinical outcomes. This methodology could be applied to other areas in medicine to assess surrogate outcomes. PMID- 19545976 TI - Can a nonequivalent choice of dosing regimen bias the results of flexible dose double blind trials? The CATIE schizophrenia trial. AB - BACKGROUND: One of the major challenges in the design of double-blind flexible dosing clinical trials comparing active drugs is the selection of dosing regimens that are equivalent across drugs. This study uses data from the CATIE schizophrenia trial to evaluate the hypothesis that drugs that were dosed somewhat higher in the trial than in typical practice would show greater efficacy and more side effects, especially at high capsule levels, than drugs that were dosed at lower relative strengths. METHODS: CATIE was a large (N=1460) randomized trial comparing 5 antipsychotics in patients with chronic schizophrenia. The blind was maintained in CATIE by prescribing identical-looking capsules of each medication. Dosing was flexible, such that PIs could prescribe from one to four capsules per day, and could modify the dose based on a patient's symptoms and side effects. Capsule strengths for olanzapine (7.5 mg) and quetiapine (200 mg) were relatively higher than for risperidone (1.5 mg), perphenazine (8 mg) or ziprasidone (40 mg). Proportional hazards models of time to all cause discontinuation and mixed regression models for continuous measures of symptoms, quality of life and side effects were used to test for interactions between randomly assigned drug and number of capsules prescribed per visit. We hypothesized that if a dosing bias was present, the flex-dosing design would result in a significant interaction such that drugs with higher relative dosing per capsule would be more effective and have more side effects than drugs with lower relative dosing and that this effect would be greatest at the largest prescribed dosing regimen (4 capsules). RESULTS: There were no significant interactions between drug assignment and number of capsules in the proportional hazards analyses of time to all cause discontinuation (p=.77, excluding ziprasidone and .74 in the ziprasidone cohort) or in the mixed model analysis of PANSS symptoms (p=.49), quality of life (p=.45); or measures of tardive dyskinesia (AIMS, p=.47). However a significant interaction was observed on the Barnes akathisia scale (p=.0005), on the Simpson Angus EPS scale (p=.10) and on the analysis of weight (p=0.014). Paired comparisons did not show the hypothesized pattern of relationships for akathisia or EPS, but such a pattern was suggested for olanzapine in the analysis of weight although it emerged at 2, 3 and 4 capsules indicating a general drug effect rather than a relative dosing difference. CONCLUSION: Dosing biases do not seem to have affected the results of the CATIE trial. PMID- 19545977 TI - Effect of risperidone on emotion recognition deficits in antipsychotic-naive schizophrenia: a short-term follow-up study. AB - BACKGROUND: Facial emotion recognition deficits [FERD] have been consistently demonstrated in treated schizophrenia patients. FERD in treatment-naive patients and the effect of antipsychotics are yet to be explored. AIMS: To examine for FERD in antipsychotic-naive schizophrenia patients and the effect of short-term atypical antipsychotic treatment on FERD. METHODS: Twenty-five antipsychotic naive schizophrenia [DSM-IV] patients and 30 age-, sex-, and education-matched healthy control subjects were assessed for FERD using the Tool for Recognition of Emotions in Neuropsychiatric DisorderS [TRENDS] - a culturally sensitive and valid tool. Psychopathology was assessed using SAPS and SANS. Performance of patients on TRENDS and psychopathology was re-assessed after short-term exposure to risperidone. RESULTS: At baseline, the patients made significantly greater errors in recognition of negative emotions of fear and disgust which improved on follow-up. This improvement was influenced by severity of baseline negative symptoms. CONCLUSION: Risperidone treatment can improve disgust recognition deficits in patients with schizophrenia. PMID- 19545978 TI - Cultural themes associated with pathways to initial use of psychiatric services in a tri-ethnic community sample of adults with schizophrenia. PMID- 19545979 TI - Changes in quality of life following cognitive-behavioral group therapy for panic disorder. AB - BACKGROUND: Data about quality of life (QoL) are important to estimate the impact of diseases on functioning and well-being. The present study was designed to assess the association of different aspects of panic disorder (PD) with QoL and to examine the relationship between QoL and symptomatic outcome following brief cognitive-behavioral group therapy (CBGT). METHOD: The sample consisted of 55 consecutively recruited outpatients suffering from PD who underwent CBGT. QoL was assessed by the Medical Outcomes Study 36-item Short-Form Health Survey (SF-36) at baseline, post-treatment and six months follow-up. SF-36 baseline scores were compared with normative data obtained from a large German population sample. RESULTS: Agoraphobia, disability, and worries about health were significantly associated with decreased QoL, whereas frequency, severity and duration of panic attacks were not. Treatment responders showed significantly better QoL than non responders. PD symptom reduction following CBGT was associated with considerable improvement in emotional and physical aspects of QoL. However, the vitality subscale of the SF-36 remained largely unchanged over time. CONCLUSIONS: Our results are encouraging for cognitive-behavior therapists who treat patients suffering from PD in groups, since decrease of PD symptoms appears to be associated with considerable improvements in QoL. Nevertheless, additional interventions designed to target specific aspects of QoL, in particular vitality, may be useful to enhance patients' well-being. PMID- 19545980 TI - Uzbekistan: psychiatry in transition. AB - The center of a national medical identity in Uzbekistan is Abu Ali Ibn Sina born in 980 AD representing Islamic medicine and scientific universalism. Psychiatric institutions were founded under Russian influence starting in the late 19th century. Today, the great challenge in psychiatry is the development of a post Soviet identity integrating Russian and Islamic traditions. PMID- 19545983 TI - Self-assembling microparticles with controllable disruption properties based on cyclodextrin interactions. AB - In this paper, we present the formation of particles by self-assembly of cyclodextrin polymers and hydrophobically modified dextran followed by a controlled disruption of the particles by addition of a trigger molecule competing for the cyclodextrin cavities. The produced particles are formed from poly(vinylpyrrolidone)-co-beta-cyclodextrin and dextran-benzoate, both biocompatible polymers, and are all in the nano-/micrometer range and hence suitable for drug delivery purposes. The particle formation was studied in different ratios of poly(vinylpyrrolidone)-co-beta-cyclodextrin and dextran benzoate by visual inspections, dynamic light scattering, isothermal titration calorimetry and SEM. The triggering of particle disruption was achieved by addition of hydroxyadamantane which has a very strong affinity towards the beta cyclodextrin cavities. The stepwise addition of hydroxyadamantane was followed by dynamic light scattering and SEM measurements, revealing a disruption of the particles due to the addition of this competitor. These particles are believed to be promising candidates for controlled drug delivery systems, due to their unique ability to disrupt in a controlled manner. PMID- 19545981 TI - An MRI-based approach for the measurement of the dorsolateral prefrontal cortex in humans. AB - The dorsolateral prefrontal cortex (DLPFC) has been implicated in the pathophysiology of mental disorders. Previous region-of-interest MRI studies that attempted to delineate this region adopted various landmarks and measurement techniques, with inconsistent results. We developed a new region-of-interest measurement method to obtain morphometric data of this region from structural MRI scans, taking into account knowledge from cytoarchitectonic postmortem studies and the large inter-individual variability of this region. MRI scans of 10 subjects were obtained, and DLPFC tracing was performed in the coronal plane by two independent raters using the semi-automated software Brains2. The intra-class correlation coefficients between two independent raters were 0.94 for the left DLPFC and 0.93 for the right DLPFC. The mean +/- S.D. DLPFC volumes were 9.23 +/- 2.35 ml for the left hemisphere and 8.20 +/- 2.08 ml for the right hemisphere. Our proposed method has high inter-rater reliability and is easy to implement, permitting the standardized measurement of this region for clinical research applications PMID- 19545982 TI - Amygdala hyperactivation in untreated depressed individuals. AB - The amygdala participates in the detection and control of affective states, and has been proposed to be a site of dysfunction in affective disorders. To assess amygdala processing in individuals with unipolar depression, we applied a functional MRI (fMRI) paradigm previously shown to be sensitive to amygdala function. Fourteen individuals with untreated DSM-IV major depression and 15 healthy subjects were studied using fMRI with a standardized emotion face recognition task. Voxel-level data sets were subjected to a multiple-regression analysis, and functionally defined regions of interest (ROI), including bilateral amygdala, were analyzed with MANOVA. Pearson correlation coefficients between amygdala activation and HAM-D score also were performed. While both depressed and healthy groups showed increased amygdala activity when viewing emotive faces compared to geometric shapes, patients with unipolar depression showed relatively more activity than healthy subjects, particularly on the left. Positive Pearson correlations between amygdala activation and HAM-D score were found for both left and right ROIs in the patient group. This study provides in vivo imaging evidence to support the hypothesis of abnormal amygdala functioning in depressed individuals. PMID- 19545984 TI - Detecting cells on the surface of a silver electrode quartz crystal microbalance using plasma treatment and graft polymerization. AB - This paper utilizes a silver electrode quartz crystal microbalance (QCM) mass sensor to detect the physiology of cells. This study also investigates the plasma surface modification of silver electrode QCMs through deposition of hexamethyldisilazane (HMDSZ) films as a protection film. To improve the cell growth, this paper also performs post-treatments by surface-grafting acrylic acid (AAc), acrylamide (AAm), and oxygen plasma treatment onto the QCM electrodes. Experimental results indicate that plasma deposition is a useful technique to protect the surface of silver electrodes. This technique extends the unpeeling time of silver electrodes from 1 to 7 days. The hydrophilic silver electrode QCM surface modified by AAm exhibited a better storage time effect than other post treatments. PMID- 19545985 TI - Surface modification of mitoxantrone-loaded PLGA nanospheres with chitosan. AB - The purpose of this research was to develop polylactic-co-glycolic acid (PLGA) nanospheres surface modified with chitosan (CS). Mitoxantrone- (MTO-) loaded PLGA nanospheres were prepared by a solvent evaporation technique. The PLGA nanospheres surface was modified with CS by two strategies (adsorption and covalent binding). PLGA nanospheres of 248.4+/-21.0 nm in diameter characterized by the laser light scattering technique, scanning electron microscopy (SEM) are spherical and its drug encapsulation efficiency is 84.1+/-3.4%. Zeta potential of unmodified nanospheres was measured to be negative -21.21+/-2.13 mV. The positive zeta potential of modified nanospheres reveals the presence of CS on the surface of the modified nanospheres. Modified nanospheres were characterized for surface chemistry by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FT-IR). FT-IR spectra exhibited peaks at 3420 cm(-1) and 1570 cm(-1), XPS spectra shows the N 1s (atomic orbital 1s of nitrogen) region of the surface of the nanospheres, corresponding to the primary amide of CS. In vitro drug release demonstrated that CS-modified nanospheres have many advantages such as prolonged drug release property and decreased the burst release over the unmodified nanospheres, and the modified nanospheres by covalent binding method could achieve the release kinetics of a relatively constant release. These data demonstrate high potential of CS-modified PLGA nanospheres for the anticancer drug carrier. PMID- 19545986 TI - Spontaneous extracutaneous systemic mastocytosis in a cynomolgus macaque (Macaca fascicularis). AB - A uniform cell population of proliferating mast cells with poor cytoplasmic granularity and a few eosinophilic infiltrates was observed in hepatic portal tracts and the cecal submucosa of an adult male cynomolgus macaque (Macaca fascicularis) that was part of a drug safety assessment toxicity study. The proliferating mast cells were positive for Giemsa and toluidine blue staining and had strong immunoreactivity for mast cell tryptase and CD68. Considering size, morphology, immunoreactivity, and distribution of the lesions a benign proliferative disorder of connective tissue mast cells was diagnosed comparable to that seen in indolent systemic mastocytosis in humans, but lacking associated skin lesions. The finding of visceral mast cell proliferation has to be included in the spontaneous background pathology spectrum of cynomolgus macaques used in toxicological studies. PMID- 19545987 TI - Biomarkers of DHA status. AB - Docosahexaenoic acid (DHA, 22:6n-3) is a long chain omega-3 fatty acid that is the primary n-3 fatty acid found in the central nervous system where it plays both a structural and functional role in cells. Because the tissues of interest are generally inaccessible for fatty acid analysis in humans and because precise DHA intake is difficult to determine, surrogate biomarkers are important for defining DHA status. Analysis of total lipid extracts or phospholipids from plasma or erythrocytes by gas chromatography meet the criteria for a useful biomarker of DHA status. Furthermore, both plasma and erythrocyte DHA levels have been correlated with brain, cardiac, and other tissue levels. Use of these biomarkers of DHA status will enable future clinical trials and observational studies to define more precisely the DHA levels required for either disease prevention or other functional benefits. PMID- 19545988 TI - Docosahexaenoic acid (DHA) and cardiovascular disease risk factors. AB - Numerous epidemiological and controlled interventional trials have supported the health benefits of long-chain omega-3 fatty acids in the form of docosahexaenoic acid (DHA, 22:6n-3) plus eicosapentaenoic acid (EPA, 20:5n-3) from fish and fish oils as well as from algal sources. The beneficial effects on cardiovascular disease and related mortality including various risk factors for cardiovascular disease (particularly lowering circulating triglyceride levels and the triglyceride:HDL-cholesterol ratio) have been observed in the absence of any concomitant blood cholesterol lowering. With appropriate dosages, consistent reductions in both fasting and postprandial triglyceride levels and moderate increases in fasting HDL-cholesterol levels have been observed with algal DHA in the majority of trials. These results are similar to findings for fish oils containing DHA and EPA. Related to greater fish intake, higher levels of DHA in circulating blood biomarkers (such as serum phospholipid) have been associated with reduced risks for the progression of coronary atherosclerosis and lowered risk from sudden cardiac death. Controlled clinical trials have also indicated the potential for algal DHA supplementation to have moderate beneficial effects on other cardiovascular disease risk factors including blood pressures and resting heart rates. Recommended intakes of DHA+EPA from numerous international groups for the prevention and management of cardiovascular disease have been forthcoming, although most have not offered specific recommendations for the optimal individual intake of DHA and EPA. PMID- 19545989 TI - The influence of pH on the leaching behaviour of inorganic components from municipal solid waste APC residues. AB - The influence of pH on the leaching behaviour of air pollution control (APC) residues produced in municipal solid waste incineration (MSWI) is addressed in this study. The residue is considered hazardous waste, and in accordance with their chemical properties, the leaching of contaminants into the environment is the main concern. Several leaching tests can be used for research studies or regulatory purposes, where a wide variety of conditions may be tested. Our work deals mainly with the leaching behaviour of toxic heavy metals (Pb, Cd, Zn, Cr, Ni, Cu) and inorganics associated with soluble salts (Na, K, Ca, Cl). The main goal is to obtain an overview of the leachability of APC residues produced in a Portuguese MSWI process. Among the different variables that may have influence on the leaching behaviour, pH of the leachant solution is the most important one, and was evaluated through pH static tests. The acid neutralization capacity (ANC) of the residue was also determined, which is in the range of 6.2-6.8 meq g(-1) (for pH=7) and 10.1-11.6 meq g(-1) (for pH=4). The analysis of the leaching behaviour is particularly important when the leaching is solubility controlled. The amphoteric behaviour of some elements was observed, namely for Pb and Zn, which is characterized through high solubilization at low and high pH and moderate or low solubility at neutral or moderate high pH. The solubility curves for Pb, Cd, Zn, Cr, Ni and Cu as a function of pH were obtained, which are very useful for predicting the leaching behaviour in different scenarios. The solubility of K and Na reveals to be nearly independent of the solution pH and the released amount is mainly availability-controlled. Moreover, the pH static test showed that Cl(-) is the most pH-independent species. The APC residue turns out to be a hazardous waste because of the high leaching of lead and chloride. On the other hand, leaching of elements like cadmium, nickel and copper is limited by the high pH of the residue, and as long as the waste keeps its ANC, the risk of mobilization of these elements is low. PMID- 19545990 TI - Effects of recycled glass substitution on the physical and mechanical properties of clay bricks. AB - In this study, wasted glasses from structural glass walls up to 45 wt.% were added into clay mixtures in brick manufacturing process. Physical and mechanical properties of clay bricks were investigated as functions of the wasted glass content and the firing temperature. The results indicated that with proper amount of wasted glasses and firing temperature, clay bricks with suitable physical and mechanical properties could be obtained. The compressive strength as high as 26 41 MPa and water absorption as low as 2-3% were achieved for bricks containing 15 30 wt.% of glass content and fired at 1100 degrees C. When the glass waste content was 45 wt.%, apparent porosity and water absorption was rapidly increased. PMID- 19545991 TI - Single-cell trapping utilizing negative dielectrophoretic quadrupole and microwell electrodes. AB - The handling of individual cells, which has attracted increasing attention, is a key technique in cell engineering such as gene introduction, drug injection, and cloning technology. Alternating current (AC) electrokinetics has shown great potential for microfluidic functions such as pumping, mixing, and concentrating particles. The non-uniform electric field gives rise to Joule heating and dielectrophoresis (DEP). The motion of particles suspended in the medium can be influenced directly, by means of dielectrophoretic effects, and indirectly, via fluid flow through a viscous drag force that affects the particles. Thus alternating current electrothermal effect (ACET) induced flow and DEP force can be combined to manipulate and trap single particles and cells. This study presents a microfluidic device which is capable of specifically guiding and capturing single particles and cells by ACET fluid flow and the negative dielectrophoretic (nDEP) trap, respectively. The experiment was operated at high frequencies (5-12 MHz) and in a culture medium whose high conductivity (sigma=1.25S/m) is of interest to biochemical analysis and environmental monitoring, which are both prone to producing ACET and nDEP. Manipulation of particle motion using ACET-induced fluid flow to the target trap is modeled numerically and is in good agreement with the experimental results. PMID- 19545992 TI - Yeast metabolic engineering for hemicellulosic ethanol production. AB - Efficient fermentation of hemicellulosic sugars is critical for the bioconversion of lignocellulosics to ethanol. Efficient sugar uptake through the heterologous expression of yeast and fungal xylose/glucose transporters can improve fermentation if other metabolic steps are not rate limiting. Rectification of cofactor imbalances through heterologous expression of fungal xylose isomerase or modification of cofactor requirements in the yeast oxidoreductase pathway can reduce xylitol production while increasing ethanol yields, but these changes often occur at the expense of xylose utilization rates. Genetic engineering and evolutionary adaptation to increase glycolytic flux coupled with transcriptomic and proteomic studies have identified targets for further modification, as have genomic and metabolic engineering studies in native xylose fermenting yeasts. PMID- 19545994 TI - A synaptic trek to autism. AB - Autism spectrum disorders (ASD) are diagnosed on the basis of three behavioral features namely deficits in social communication, absence or delay in language, and stereotypy. The susceptibility genes to ASD remain largely unknown, but two major pathways are emerging. Mutations in TSC1/TSC2, NF1, or PTEN activate the mTOR/PI3K pathway and lead to syndromic ASD with tuberous sclerosis, neurofibromatosis, or macrocephaly. Mutations in NLGN3/4, SHANK3, or NRXN1 alter synaptic function and lead to mental retardation, typical autism, or Asperger syndrome. The mTOR/PI3K pathway is associated with abnormal cellular/synaptic growth rate, whereas the NRXN-NLGN-SHANK pathway is associated with synaptogenesis and imbalance between excitatory and inhibitory currents. Taken together, these data strongly suggest that abnormal synaptic homeostasis represent a risk factor to ASD. PMID- 19545993 TI - Epigenetic influences on brain development and plasticity. AB - A fine interplay exists between sensory experience and innate genetic programs leading to the sculpting of neuronal circuits during early brain development. Recent evidence suggests that the dynamic regulation of gene expression through epigenetic mechanisms is at the interface between environmental stimuli and long lasting molecular, cellular and complex behavioral phenotypes acquired during periods of developmental plasticity. Understanding these mechanisms may give insight into the formation of critical periods and provide new strategies for increasing plasticity and adaptive change in adulthood. PMID- 19545995 TI - What can developmental and comparative cognitive neuroscience tell us about the adult human brain? PMID- 19545996 TI - Clinical benefit in oncology trials: is this a patient-centred or tumour-centred end-point? AB - BACKGROUND: Clinical benefit (CB) was first successfully used as an end-point in 1997 in the pivotal study of gemcitabine in advanced pancreas cancer. In the trial by Burris et al. CB was a composite measure of pain, performance status and weight. Here we describe how CB has been used in oncology trials since that time. METHODS: We performed an electronic search (www.jco.org) for reports of all clinical trials (phase I, II and III) published in the Journal of Clinical Oncology 1997-2008 citing 'clinical benefit'. Eligible trials were those reporting clinical benefit as an end-point. Details related to study methodology, sponsorship and end-points were abstracted. Use of CB was classified as patient centred if it referred to improvement in the clinical parameters used by Burris et al. or in other disease-related symptoms. CB was classified as tumour centred if it related to objective tumour criteria for partial/complete response and/or stable disease. Descriptive statistics were used to summarise findings and the chi-square test was used to compare proportions. RESULTS: Seventy-one trials reporting CB as an end-point were identified: 37 in breast, 8 in pancreas and 26 in other cancers. The definition of CB was patient centred in 20 trials (28%) and tumour centred in 51 trials (72%). Only 20% (14/71) of trials (including all 8 pancreas studies) used the original Burris definition. Among the 71 trials reporting clinical benefit, in only 31 (44%) cases was the end-point defined as a primary or secondary study objective. Trials with a patient-centred definition of CB were considerably more likely to do so than trials with a tumour-centred definition (19/20, 95% versus 12/51, 24%, p<0.0001). Study variables associated with the use of a tumour-centred definition include: disease site (breast 35/37, 95%; all others 16/34, 47%, p<0.001) and intervention (hormone or targeted agent 38/40, 95%; chemotherapy 13/31, 42%, p<0.001). There has been a steady increase in the number of trials using CB as an end-point; in the second half of the study period the number of trials increased from 17 to 54, along with the proportion of trials with a tumour-centred definition (10/17, 59% to 41/54, 76%, p=0.09). CONCLUSIONS: Despite its initial definition, clinical benefit is often used to describe objective tumour findings. Clinical trials should use end-points in a consistent manner to enable clear communication between investigators, clinicians and patients about the benefit of novel therapies. PMID- 19545997 TI - Meta-analysis of risk factors for cutaneous melanoma according to anatomical site and clinico-pathological variant. AB - A systematic meta-analysis was performed to evaluate if cutaneous melanoma (CM) risk factors differ depending on body site and histological type. Adjusted estimates were extracted from 24 observational studies, for a total of 16,180 cases. Multivariate random-effects models were used to obtain summary relative risk (RR) estimates for all risk factors by body site and histological type. Summary RRs suggest that high naevus counts are strongly associated with CM on usually not sun exposed sites (p<0.001) while different patterns of sun exposure show a tendency for higher RRs for CM on usually sun exposed sites than on other body sites (p=0.087). Continuous pattern was found to be significantly inversely associated with CM for unexposed sites (p=0.01). RRs also differed by body site for skin (p=0.01) and hair colour (p=0.01), and these differences could be attributed to gene variability. This finding seems to suggest different aetiologic pathways of melanoma development by anatomical site. PMID- 19545998 TI - Continuous hydrogen and butyric acid fermentation by immobilized Clostridium tyrobutyricum ATCC 25755: effects of the glucose concentration and hydraulic retention time. AB - The effects of the hydraulic retention time (HRT=8, 10, 12 or 16.7 h) and glucose concentration (30, 40 or 50 g/L) on the production of hydrogen and butyrate by an immobilized Clostridium tyrobutyricum culture, grown under continuous culturing conditions, were evaluated. With 30 g/L glucose, the higher HRTs tested led to greater butyrate concentrations in the culture, i.e., 9.3 g/L versus 12.9 g/L with HRTs of 8 h and 16.7 h, respectively. In contrast, higher biogas and hydrogen production rates were generally seen when the HRT was lower. Experiments with different glucose concentrations saw a significant amount of glucose washed out when 50 g/L was used, the highest being 22.7 g/L when the HRT was 16.7 h. This study found the best conditions for the continuous production of hydrogen and butyric acid by C. tyrobutyricum to be with an HRT of 12 h and a glucose concentration of 50 g/L, respectively. PMID- 19546000 TI - Bioenergy from permanent grassland--a review: 2. Combustion. AB - The aim of this review is to summarize current knowledge on suitability and sustainability of grassland biomass for combustion. In the first section grassland management for solid biofuel as well as information on harvest, postharvest and firing technology are described. An extensive grassland management system with one late cut and low level of fertilization is favored for grass as a solid biofuel. The grass harvest usually involves drying in the field and clearing with conventional farm machinery. Pelleting or briquetting improves the biofuel quality. Grass combustion is possible as stand-alone biomass-firing or co-firing with other fuels. Firing herbaceous biomass requires various specific adaptations of the different combustion technologies. In the second section economic and environmental aspects are discussed. Costs for biomass supply mainly depend on yields and harvesting technologies, while combustion costs are influenced by the size and technical design of the plant. Market prices for grass and possible subsidies for land use are crucial for profitability. Regarding biogeochemical cycles a specific feature of combustion is the fact that none of the biomass carbon and nitrogen removed at harvest is available for return to the grassland. These exports can be compensated for by fixation from the air given legumes in the vegetation and sufficient biomass production. Greenhouse gas emissions can be considerably reduced by grass combustion. Solid biofuel production has a potential for predominantly positive impacts on biodiversity due to the extensive grassland management. PMID- 19545999 TI - The effect of biological pretreatment with the selective white-rot fungus Echinodontium taxodii on enzymatic hydrolysis of softwoods and hardwoods. AB - Selective white-rot fungi have shown potential for lignocellulose pretreatment. In the study, a new fungal isolate, Echinodontium taxodii 2538, was used in biological pretreatment to enhance the enzymatic hydrolysis of two native woods: Chinese willow (hardwood) and China-fir (softwood). E. taxodii preferentially degraded the lignin during the pretreatment, and the pretreated woods showed significant increases in enzymatic hydrolysis ratios (4.7-fold for hardwood and 6.3-fold for softwood). To better understand effects of biological pretreatment on enzymatic hydrolysis, enzyme-substrate interactions were investigated. It was observed that E. taxodii enhanced initial adsorption of cellulase but which did not always translate to high initial hydrolysis rate. However, the rate of change in hydrolysis rate declined dramatically with decreasing irreversible adsorption of cellulase. Thus, the enhancement of enzymatic hydrolysis was attributed to the decline of irreversible adsorption which may result from partial lignin degradation and alteration in lignin structure after biological pretreatment. PMID- 19546001 TI - Bioenergy from permanent grassland--a review: 1. Biogas. AB - Grassland biomass is suitable in numerous ways for producing energy. It is well established as feedstock for biogas production. The aim of this review is to summarize current knowledge on suitability and sustainability of grassland biomass for anaerobic digestion. In the first section grassland management for biogas feedstock as well as specifics of harvest, postharvest and digestion technology are described. Methane yields from grass are influenced by many factors. While the effects of some parameters such as grass species, cutting period and management intensity can be regarded as well known, other parameters such as preservation and processing still need investigation. In the second section economic aspects and environmental impacts are discussed. Profitability can be achieved depending on grass silage supply costs and the concept of anaerobic digestion and energy use. Grassland biomass for biogas production competes with other feedstock and other forms of grassland use, in particular animal husbandry. In developed countries a growing production of milk and meat is achieved with decreasing ruminant numbers, resulting in an increasing amount of surplus grassland with a remarkable bioenergy potential. In emerging and developing countries a rapidly rising demand for and production of milk and meat induce growing pressure on grasslands, so that their use for animal feed presumably will take priority over use for bioenergy. Grasslands provide a variety of essential environmental benefits such as carbon storage, habitat function, preservation of ground and surface water quality. When producing biogas from grassland these benefits will remain or even grow, providing appropriate grassland management is implemented. In particular, greenhouse gas emissions can be considerably reduced. PMID- 19546002 TI - The kinetics of nicotine degradation, enzyme activities and genotoxic potential in the characterization of tobacco waste composting. AB - This study aimed to determine nicotine biodegradation and the genotoxic potential of nicotine and its degradation products during the process of tobacco waste composting. Composting was carried out using two methods, i.e. the addition of 20% (bioreactor A) or 40% tobacco wastes to sewage sludge (bioreactor B) and control--sewage sludge (bioreactor C). Wheat straw was used as a structure forming material. As a result of composting the contents of C and N in the bioreactors changed, the C:N ratio in bioreactor A changed from 22.8 to 13.00, and that in bioreactor B changed from 23.5 to 12.00. After composting, the biodegradation rate of nicotine was 78% in bioreactor A and 80% in bioreactor B, respectively. Using the Ames test it was shown that the composts produced did not exhibit mutagenicity. PMID- 19546004 TI - Real-time fluorescence monitoring of GSK3beta-catalyzed phosphoryation by use of a BODIPY-based Zn(II)-Dpa chemosensor. AB - We developed a new fluorescence assay system for GSK3beta-catalyzed kinase reaction using the BODIPY-based fluorescent chemosensor. This system exploits the selective sensing property of the chemosensor for a (i, i+4) bis-phosphorylated peptide, which allows us to conveniently detect the phosphorylation reaction with a fluorescence increase in a real-time fashion. PMID- 19546003 TI - Use of anionic polymer, poly(methyl vinyl ether-maleic anhydride)-coated beads for capture of respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) is the single most important cause of severe lower respiratory tract infections in infants and young children, and a major public health concern in pediatrics. However, current diagnostic methods for RSV are not sufficiently sensitive. In addition, there is no simple method for enhancing RSV detection. Here, a method for capturing RSV from nasal fluid has been developed using magnetic beads coated with an anionic polymer, poly(methyl vinyl ether-maleic anhydrate). The beads were incubated with RSV-infected nasal fluid, then separated from the supernatant by applying a magnet field and washed. The adsorption [corrected] of RSV by the beads was confirmed by immunochromatography, reverse transcription-polymerase chain reaction, Western blotting and an enzyme-linked immunosorbent assay, which indicated the presence of nucleocapsid protein, fusion protein, and the viral genome of RSV on the incubated beads. Therefore, this capture method will contribute to the improvement of RSV detection. PMID- 19546005 TI - When is the appropriate time for surgical intervention of the herniated lumbar disc in the adolescent? AB - Symptomatic lumbar disc herniation in the adolescent is uncommon. The appropriate treatment in this particular age group is not clear. We conducted a retrospective review of the medical, surgical, and radiological records of six adolescents with symptomatic lumbar disc herniation who underwent microdiscectomy after failed conservative therapy. The mean follow-up was 13 months. All patients improved quickly and returned to their normal activity levels. We suggest that severe pain resulting from a herniated lumbar disc, even without any neurological deficit, is an indication for microdiscectomy in adolescents so that these patients return to full-time education and normal activities as soon as possible. PMID- 19546006 TI - SitePredicting the cleavage of proteinase substrates. AB - Proteinases are enzymes that play important roles in vital cellular and extracellular processes by hydrolytically cleaving peptide bonds in their protein substrates. This cleavage can be non-specific as part of degradation during protein catabolism or highly specific as part of proteolytic cascades and signal transduction events. Several web tools are available for predicting possible cleavage sites in candidate substrates. Here, we compare existing prediction tools with SitePrediction, a novel and user-friendly tool for identifying potential cleavage sites. This prediction is based on known datasets found in the literature, stored in web-accessible repositories or generated by our own experiments. Comparison of the different programs shows that SitePrediction makes it possible to derive more reliable predictions. In addition, this tool allows the use of a wide range of proteinases. PMID- 19546007 TI - Earthquake precursory studies in Kangra valley of North West Himalayas, India, with special emphasis on radon emission. AB - The continuous soil gas radon monitoring is carried out at Palampur and the daily monitoring of radon concentration in water is carried out at Dharamshala region of Kangra valley of North West Himalayas, India, a seismic zone V, to study the correlation of radon anomalies in relation to seismic activities. In this study, radon monitoring in soil was carried out by using barasol probe manufactured by Algade France, whereas the radon content in water was recorded using RAD 7 radon monitoring system of Durridge Company USA. The effect of meteorological parameters viz. temperature, pressure, wind velocity, rainfall, and humidity on radon emission has been studied. The seasonal average value and standard deviation of radon in soil and water is calculated to find the radon anomaly to minimize the effect of meteorological parameters on radon emission. The radon anomalies observed in the region have been correlated with the seismic events of M>or=2 reported by Wadia Institute of Himalayas Geology Dehradoon and Indian Meteorological Department, New Delhi in NW Himalayas within 250km distance from the monitoring stations. PMID- 19546008 TI - Clinical relevance of contrast-enhanced ultrasound in monitoring anti-angiogenic therapy of cancer: current status and perspectives. AB - Angiogenesis regulation is one of the newest fronts in the fight against cancer. Anti-angiogenic therapy is based on inhibiting factors required to solicit vessel formation thus cutting-off the tumor's supply of nutriments and oxygen. Initial vascular response is followed by formation of necrosis. Volumetric regression occurs more tardively. Effective monitoring of this new therapeutic approach thus requires imaging techniques that can detect early microvascular changes. A number of clinical studies provide evidence that contrast-enhanced ultrasound (CEUS) can provide early indication of tumor response to anti-angiogenic therapy. More sophisticated imaging and analysis techniques for CEUS and contrast agents targeted for adhesion to anti-angiogenic markers have also demonstrated promise in animal model studies. This review underlines the relevance of CEUS for anti angiogenic therapy monitoring by summarizing the current clinical results, emerging CEUS techniques and preclinical data. PMID- 19546009 TI - Bone as an endocrine organ. AB - Although bone has long been recognized as a target for hormones influencing calcium and phosphorus homeostasis and bone structure, recent evidence shows that the skeleton itself produces at least two hormones, fibroblast growth factor 23 (FGF23) and osteocalcin. FGF23 is produced by osteocytes in bone and acts on the kidney to inhibit 1alpha-hydroxylation of vitamin D and promote phosphorus excretion. Mouse genetics studies revealed that the osteoblast product, osteocalcin, acts on the pancreatic beta-cell to enhance insulin production and on peripheral tissues to increase glucose utilization as a result of increased insulin sensitivity and to reduce visceral fat. This review highlights the recent studies indicating bone's role as an endocrine organ. PMID- 19546011 TI - Visual perspective and genetics: a commentary on Lemogne and colleagues. AB - Lemogne and colleagues offer an interesting extension to their previous work on visual perspective and depression: Individuals at-risk for depression (defined as higher scores on Harm Avoidance), without a history of mood disorders, report retrieval of positive memories from the 3rd person perspective. Their findings suggest that the retrieval of positive experiences from the 3rd person perspective may be a risk-factor for depression, not just a lingering consequence of it. Their study, however, also reports a genetic association in a severely underpowered sample. Rather than focusing on gene x environment interactions, which large, well-powered studies on related phenotypes have failed to detect, a greater understanding of the phenomenology of visual perspective may be a more fruitful avenue for future research. PMID- 19546012 TI - Evaluation of glenoid capsulolabral complex insertional anatomy and restoration with single- and double-row capsulolabral repairs. AB - BACKGROUND: The purpose of this study was to evaluate the normal glenoid insertional anatomy of the anterior-inferior capsulolabral complex and to compare the ability of a single-row repair and a double-row suture bridge repair to restore the insertional anatomy. METHODS: Eight fresh frozen cadaver shoulders were dissected and the native glenoid insertion of the anterior-inferior capsulolabral complex was digitized. Bankart lesions were created, the shoulders were randomized to receive either the standard single-row suture anchor repair or a double-row suture bridge repair, and the insertion repair sites were then digitized. RESULTS: The single-row repair recreated 42.3% of the native footprint surface area while the double-row repair recreated 85.9%. The double-row repair was significantly larger and recreated significantly more of the native footprint compared with single-row repair (P < .01). CONCLUSION: Double-row repair of the capsulolabral complex reestablishes the native insertional footprint on the anterior inferior glenoid better than a single-row repair. LEVEL OF EVIDENCE: Basic science study. PMID- 19546010 TI - Structural analysis of sulfated glycans by sequential double-permethylation using methyl iodide and deuteromethyl iodide. AB - MALDI mass spectrometric characterization of sulfated glycans is often challenging due to their low ionization response in the positive ion mode. Here we demonstrate a new analytical approach, allowing the measurement of sulfated glycans by substituting the sulfate group with a deuteromethyl group. Sulfated glycan samples are initially permethylated before the methanolytic cleavage of their sulfate groups. Desulfated and permethylated glycans are then subjected to another permethylation step using deuteromethyl iodide to label the hydroxyl groups resulting from methanolysis. The number of attached sulfate groups is subsequently calculated from the mass-shift resulting from the chemical cleavage of these sulfate groups. The position of the sulfate substitution is then determined by collision-induced dissociation (CID) tandem mass spectrometry of permethylated and permethylated plus deuteromethylated samples. The described approach was initially optimized and validated using linear standard glycans, while its effectiveness has also been demonstrated here for the N-glycans derived from bovine thyroid-stimulating hormone (bTSH). PMID- 19546013 TI - Rasmussen's encephalitis: experience from a developing country based on a group of medically and surgically treated patients. AB - PURPOSE: To describe the attributes of patients with Rasmussen's encephalitis (RE) seen in a tertiary epilepsy referral center in southern India and to enquire factors helpful in predicting responsiveness to immunotherapy. METHODS: We diagnosed RE based on the European consensus criteria. To assess the factors that could potentially predict the natural course and therapeutic outcome, we subcategorized our patients according to age at onset ( 6 years), duration from onset to presentation ( 2 years), immunotherapy versus surgery, and early ( or = 6.23 WU being the optimal threshold after adjustment for age, gender, composite physiological index (CPI) and diagnosis of idiopathic pulmonary fibrosis (OR 11.09; 95% CI 2.54 to 48.36; p = 0.001). Early mortality was linked, albeit less strongly, to right ventricular dilation at echocardiography, but not to other non-invasive variables or mPAP. Overall mortality was most strongly associated with increasing CPI levels. Correlations between PVR and non-invasive variables were moderate (R(2) <0.32), improving little following construction of a multivariate index which did not itself predict mortality. CONCLUSION: In severe DLD, early mortality is strongly linked to increased PVR but not to other RHC or non-invasive variables. These findings suggest that the threshold for RHC in severe DLD should be low, enabling prioritisation of aggressive treatment including lung transplantation. PMID- 19546097 TI - Oral treatment with a Brachystemma calycinum D don plant extract reduces disease symptoms and the development of cartilage lesions in experimental dog osteoarthritis: inhibition of protease-activated receptor 2. AB - OBJECTIVE: The aims of this study were to evaluate the effect of oral treatment with a whole plant extract of Brachystemma calycinum D don (BCD) on the development of osteoarthritic lesions and symptoms in the experimental dog anterior cruciate ligament (ACL) transection model and to document its mechanism of action. METHODS: Osteoarthritis was induced by sectioning the ACL of the right knee in crossbred dogs. There were two experimental groups (n=6-7 dogs/group): placebo and BCD extract (200 mg/kg per day) given orally for 8 weeks. Macroscopic and histopathological evaluation of cartilage lesions and immunohistochemical analysis of cartilage to assess levels of inducible nitric oxide synthase (iNOS), matrix metalloprotease 13 (MMP-13) and protease activated receptor 2 (PAR-2) were done. A gait analysis of dogs was performed. RESULTS: Treatment with BCD reduced the severity (depth) (p=0.04) and histopathological score (p<0.02) of osteoarthritis cartilage lesions. BCD treatment also significantly reduced the osteoarthritis chondrocyte level of key inflammatory and catabolic factors (iNOS, p=0.009 and MMP-13, p=0.003) as well as the level of PAR-2 (p=0.03). Dogs treated with BCD showed a significant improvement in peak vertical force measured at 8 weeks (p<0.05). CONCLUSIONS: Treatment with BCD extract exerts a positive effect on the prevention of cartilage lesions induced by joint instability, and improves joint function. This effect was associated with the inhibition of major catabolic and inflammatory mediators. This study is the first to demonstrate that a therapeutic intervention that can inhibit PAR-2 is associated with a disease modifying osteoarthritis effect. PMID- 19546098 TI - Non-steroidal anti-inflammatory drugs in sports medicine: guidelines for practical but sensible use. AB - INTRODUCTION: Non-steroidal anti-inflammatory drugs (NSAID) are commonly used in sports medicine. NSAID have known anti-inflammatory, analgesic, antipyretic and antithrombotic effects, although their in-vivo effects in treating musculoskeletal injuries in humans remain largely unknown. NSAID analgesic action is not significantly greater than paracetamol for musculoskeletal injury but they have a higher risk profile, with side-effects including asthma exacerbation, gastrointestinal and renal side-effects, hypertension and other cardiovascular diseases. DISCUSSION: The authors recommend an approach to NSAID use in sports medicine whereby simple analgesia is preferentially used when analgesia is the primary desired outcome. However, based both on the current pathophysiological understanding of most injury presentations and the frequency that inflammation may actually be a component of the injury complex, it is premature to suppose that NSAID are not useful to the physician managing sports injuries. The prescribing of NSAID should be cautious and both situation and pathology specific. Both dose and duration minimisation should be prioritized and combined with simple principles of protection, rest, ice, compression, elevation (PRICE), which should allow NSAID-sparing. NSAID use should always be coupled with appropriate physical rehabilitation. CONCLUSION: NSAID are probably most useful for treating nerve and soft-tissue impingements, inflammatory arthropathies and tenosynovitis. They are not generally indicated for isolated chronic tendinopathy, or for fractures. The use of NSAID in treating muscle injury is controversial. Conditions in which NSAID use requires more careful assessment include ligament injury, joint injury, osteoarthritis, haematoma and postoperatively. PMID- 19546100 TI - Sonographic long-term study: paediatric growth charts for single kidneys. AB - AIMS: To explore the clinical course of children with "single kidney" (defined as either a solitary or single functioning kidney) with reference to renal function (glomerular filtration rate (GFR) and proteinuria), body height and particularly sonomorphological features. PATIENTS AND METHODS: This retrospective monocentric study evaluated 119 children with a solitary or single functioning kidney (>90% unilateral function on isotope scan) between 1997 and 2007. Patients were followed for 6.3 years (median, range 1-17) and had at least three renal ultrasound examinations (median 8). During recruitment six children were identified with chronic kidney disease (CKD) stage III or worse. These patients were analysed separately. RESULTS: The aetiology of "single kidney" was attributed to contralateral multicystic dysplastic kidney (26%), tumournephrectomy (24%), renal agenesis (18%), hypo/dysplasia (11%) and obstructive or refluxive uropathy (18%). Irrespective of aetiology, the sonographic dimensions of "single kidneys" were in the upper range of normal paired kidneys and showed adequate growth. Compensatory renal hypertrophy (defined as >95% CI on two or more recent measurements) occurred in a third of patients. All six patients with CKD and GFR less than 60 ml/minute per 1.73 m(2) had pathological sonomorphology of their "single kidney" with inadequate renal growth (6/6), abnormal echogenicity (5/6), hypo/dysplasia (5/6). In addition, proteinuria (5/6) and short stature (3/6) were found. CONCLUSIONS: New reference centiles were generated to assess renal size of "single kidneys" in paediatric patients. These charts will facilitate counselling of patients and parents. Further evidence for a benign clinical course of children with "single kidney" and absent additional pathology of the remnant kidney is presented. PMID- 19546099 TI - A de novo 1p34.2 microdeletion identifies the synaptic vesicle gene RIMS3 as a novel candidate for autism. AB - BACKGROUND: A child with autism and mild microcephaly was found to have a de novo 3.3 Mb microdeletion on chromosome 1p34.2p34.3. The hypothesis is tested that this microdeletion contains one or more genes that underlie the autism phenotype in this child and in other children with autism spectrum disorders. METHODS: To search for submicroscopic chromosomal rearrangements in the child, array comparative genomic hybridisation (aCGH) was performed using a 19 K whole genome human bacterial artificial chromosome (BAC) array and the Illumina 610-Quad BeadChip microarray. Ingenuity pathway analysis (IPA) was used to construct functional biological networks to identify candidate autism genes. To identify putative functional variants in candidate genes, mutation screening was performed using polymerase chain reaction (PCR) based Sanger sequencing in 512 unrelated autism patients and 462 control subjects. RESULTS: A de novo 3.3 Mb deletion containing approximately 43 genes in chromosome 1p34.2p34.3 was identified and subsequently confirmed using fluorescence in situ hybridization (FISH). Literature review and bioinformatics analyses identified Regulating Synaptic Membrane Exocytosis 3 (RIMS3) as the most promising autism candidate gene. Mutation screening of this gene in autism patients identified five inherited coding variants, including one (p.E177A) that segregated with the autism phenotype in a sibship, was predicted to be deleterious, and was absent in 1161 controls. CONCLUSIONS: This case report and mutation screening data suggest that RIMS3 is an autism causative or contributory gene. Functional studies of RIMS3 variants such as p.E177A should provide additional insight into the role of synaptic proteins in the pathophysiology of autism. PMID- 19546101 TI - Frequent medical absences in secondary school students: survey and case-control study. AB - OBJECTIVE: To determine the prevalence of frequent absence (>20% of the school year) for reasons recorded as "medical" in secondary schools; to test the hypothesis that it is associated with physical symptoms and psychiatric disorder and not with serious organic disease; to assess unmet need for psychiatric management. DESIGN: Survey using routinely collected data and case-control study SETTING: Local authority secondary schools in Edinburgh, UK. PARTICIPANTS: School students in the first 4 years of secondary school: cases were those with frequent medical absence and controls those with a good attendance record (best 10% of year group), matched for age, gender and school class. MEASURES: Period prevalence of frequent absences. Cases and controls (students and their parents) completed questionnaires about the students' symptoms. Students were given a psychiatric diagnostic interview and a medical examination. The records of specialist medical services used by the students were reviewed. RESULTS: A substantial minority (2.2%) of students had frequent medical absences. Only seven of 92 (8%) cases had a serious organic disease and 10 of 92 (11%) had symptom defined syndromes; the remainder had physical symptoms and minor medical illness. Frequent medical absence was strongly associated with psychiatric disorder (45% in cases vs 17% in controls, p<0.001, 95% CI for odds ratio 1.37 to 4.02). Only 14 of the 41 cases (34%) with a psychiatric diagnosis had attended NHS psychiatric services. CONCLUSIONS: Frequent absence for medical reasons is common, and more comprehensive management, including psychiatric assessment, is required to prevent long-term adverse consequences. PMID- 19546102 TI - The importance of nurse-led home visits in the assessment of children with problematic asthma. AB - OBJECTIVE: To evaluate and identify potentially modifiable factors in children with problematic asthma by a nurse-led assessment and home visit. DESIGN: Observational cohort study. SETTING: A tertiary paediatric respiratory centre. PATIENTS: 71 children, aged 4.5-17.5 years, with problematic asthma currently under follow-up at a tertiary respiratory centre. INTERVENTIONS: A nurse-led hospital visit followed by a home visit. MAIN OUTCOME MEASURES: Identification and attempted change of exacerbating factors so that further investigations and consideration of off-label, potentially toxic, asthma therapies were not necessary. RESULTS: Potentially modifiable factors were identified in 56 (79%) children. Many children had multiple causes for poor control. The most important were ongoing allergen exposure, 22 children (31%); passive or active smoking, 18 children (25%); medication issues including adherence, 34 children (48%); psychosocial factors, 42 families (59%). The home visit contributed valuable information to this assessment. At the home visit house dust mite avoidance measures were found to be inadequate in 84% of those sensitised; medications were not easily available for inspection or were out of date in 23%; 74% of psychology referrals were made after the home visit. In 39 children (55%) the factors identified and the interventions recommended meant that further escalation of treatment was avoided. CONCLUSIONS: Nurse-led assessments including a home visit can help identify potentially modifiable factors for poorly controlled symptoms in children with problematic asthma. PMID- 19546104 TI - A diffusion tensor MRI study of cervical cord damage in benign and secondary progressive multiple sclerosis patients. AB - BACKGROUND: Diffusion tensor (DT) MRI enables quantification of the severity of brain and cervical cord pathology in multiple sclerosis (MS). OBJECTIVE: To investigate DT MRI patterns of cervical cord damage in patients with benign MS (BMS) and secondary progressive MS (SPMS), in order to achieve a better understanding of the mechanisms underlying the development of irreversible disability in MS. METHODS: Conventional and DT MRI scans of the cervical cord and brain were acquired from 40 BMS patients, 28 SPMS patients and 18 healthy individuals. Cervical cord and brain mean diffusivity (MD) and fractional anisotropy (FA) maps were created and average MD and FA were calculated. Cross sectional cord area (CSA) was also computed. RESULTS: 37 (92%) BMS patients and all (100%) SPMS patients had macroscopic cervical cord lesions. Compared with healthy individuals, BMS patients had higher average cord MD while SPMS patients had higher average cord MD, lower average cord FA and lower average CSA. Compared with BMS patients, SPMS patients had lower cord average FA and lower average CSA. In MS patients, Expanded Disability Status Scale (EDSS) was correlated with CSA (r = -0.47, p<0.0001), average cord FA (r = -0.37, p = 0.002) and brain T2 lesion volume (LV) (r = 0.34, p = 0.005). A multivariate regression model identified CSA, average cord FA and brain T2 LV as variables independently influencing the EDSS score (r = 0.58, p<0.0001). CONCLUSIONS: Cervical cord damage outside focal macroscopic lesions is limited in patients with BMS. The assessment of cord and brain pathology provides complementary information to improve the understanding of disability accumulation in MS. PMID- 19546103 TI - Exposure-response relationship between lung cancer and polycyclic aromatic hydrocarbons (PAHs). AB - OBJECTIVES: To estimate the exposure-response function associating polycyclic aromatic hydrocarbon (PAH) exposure and lung cancer, with consideration of smoking. METHODS: Mortality, occupational exposure and smoking histories were ascertained for a cohort of 16,431 persons (15,703 men and 728 women) who had worked in one of four aluminium smelters in Quebec from 1950 to 1999. A variety of exposure-response functions were fitted to the cohort data using generalised relative risk models. RESULTS: In 677 lung cancer cases there was a clear trend of increasing risk with increasing cumulative exposure to PAH measured as benzo(a)pyrene (BaP). A linear model predicted a relative risk of 1.35 (95% CI 1.22 to 1.51) at 100 microg/m(-3) BaP years, but there was a significant departure from linearity in the direction of decreasing slope with increasing exposures. Among the models tried, the best fitting were a two-knot cubic spline and a power curve (RR = (1+bx)(p)), the latter predicting a relative risk of 2.68 at 100 microg/m(-3) BaP years. Additive models and multiplicative models for combining risks from occupational PAH and smoking fitted almost equally well, with a slight advantage to the additive. CONCLUSION: Despite the large cohort with long follow-up, the shape of the exposure-response function and the mode of combination of risks due to occupational PAH and smoking remains uncertain. If a linear exposure-response function is assumed, the estimated slope is broadly in line with the estimate from a previous follow-up of the same cohort, and somewhat higher than the average found in a recent meta-analysis of lung cancer studies. PMID- 19546106 TI - Epidemiology and pathophysiology of falls in facioscapulohumeral disease. AB - BACKGROUND AND AIMS: Muscle weakness is a potentially important, yet poorly studied, risk factor for falls. Detailed studies of patients with specific myopathies may shed new light on the relation between muscle weakness and falls. Here falls in patients with facioscapulohumeral disease (FSHD) who suffered from lower limb muscle weakness were examined. This study provides insights into the prevalence, relevance and pathophysiology of falls in FSHD. METHODS: A validated questionnaire was used as well as a prospective 3 month follow-up to examine the prevalence, circumstances and consequences of falls in 73 patients with FSHD and 49 matched healthy controls. In a subgroup of 28 subjects, muscle strength was also examined and balance was assessed electrophysiologically using body worn gyroscopes. RESULTS: In the questionnaire, 30% of the patients reported falling at least once a month whereas none of the controls did. Injuries occurred in almost 70% of the patients. The prospective study showed that patients fell mostly at home, mainly due to intrinsic (patient related) causes, and usually in a forward direction. Fallers were unstable while climbing stairs, rising from a chair and standing with eyes closed whereas non-fallers had normal balance control. Frequent fallers had greater muscle weakness than infrequent fallers. CONCLUSION: These findings demonstrate the high prevalence and clinical relevance of falls in FSHD. The relation between muscle weakness and instability among fallers is also highlighted. Because patients fell mainly at home, fall prevention strategies should focus on home adaptations. As mainly intrinsic causes underlie falls, the impact of adopting balance strategies or balance training should be explored in this patient group. PMID- 19546105 TI - MR spectroscopy indicates diffuse multiple sclerosis activity during remission. AB - OBJECTIVE: To test the hypothesis that diffuse abnormalities precede axonal damage and atrophy in the MRI normal-appearing tissue of relapsing-remitting (RR) multiple sclerosis (MS) patients, and that these processes continue during clinical remission. METHODS: Twenty-one recently diagnosed mildly disabled (mean disease duration 2.3 years, mean Expanded Disability Status Scale score of 1.4) RR MS patients and 15 healthy matched controls were scanned with MRI and proton MR spectroscopic imaging ((1)H-MRSI) at 3 T. Metabolite concentrations: N acetylaspartate (NAA) for neuronal integrity; choline (Cho) for membrane turnover rate; creatine (Cr) and myo-inositol (mI) for glial status were obtained in a 360 cm(3) volume of interest (VOI) with 3D multivoxel (1)H-MRSI. They were converted into absolute amounts using phantom replacement and normalised into absolute concentrations by dividing by the VOI tissue volume fraction obtained from MRI segmentation. RESULTS: The patients' mean VOI tissue volume fraction, 0.92 and NAA concentration, 9.6 mM, were not different from controls' 0.94 and 9.6 mM. In contrast, the patients' mean Cr, Cho and mI levels 7.7, 1.9 and 4.1 mM were 9%, 14% and 20%, higher than the controls' 7.1, 1.6 and 3.4 mM (p = 0.0097, 0.003 and 0.0023). CONCLUSIONS: The absence of early tissue atrophy and apparent axonal dysfunction (NAA loss) in these RR MS patients suggests that both are preceded by diffuse glial proliferation (astrogliosis), as well as possible inflammation, demyelination and remyelination reflected by elevated mI, Cho and Cr, even during clinical remission and despite immunomodulatory treatment. PMID- 19546108 TI - Prediction of conversion from clinically isolated syndrome to clinically definite multiple sclerosis according to baseline MRI findings: comparison of revised McDonald criteria and Swanton modified criteria. AB - BACKGROUND: The International Panel on the Diagnosis of Multiple Sclerosis first incorporated abnormalities demonstrated by brain and spinal cord MRI into the diagnostic criteria (McDonald criteria) for multiple sclerosis (MS), which were later revised in 2005. In 2006, Swanton and colleagues modified the MRI criteria to simplify and speed the diagnosis. OBJECTIVE: The purpose of this study was to compare the ability of two sets of criteria (the revised McDonald MRI criteria and Swanton's modified criteria) to predict conversion from clinically isolated syndrome (CIS) to clinically definite multiple sclerosis (CDMS) from baseline MRI findings. METHODS: Sixty-four patients presenting with CIS suggestive of multiple sclerosis were recruited from 2001 to 2006 and followed up for at least 2 years. Their baseline brain and spinal cord MRI studies were retrospectively evaluated. The patients who developed CDMS during follow-up were treated as positive cases. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of the two MRI dissemination-in-space criteria were calculated. RESULTS: Thirty patients (46.9%) converted to CDMS. The sensitivity specificity, PPV, NPV and accuracy (%) of the revised McDonald criteria were 53, 100, 100, 71 and 78, respectively, while those for Swanton's modified criteria were 60, 100, 100, 74 and 81. CONCLUSIONS: In conclusion, Swanton's modified criteria are more sensitive and accurate (but not significantly so). However, Swanton's criteria are simpler to use and have equally high specificity and PPV. PMID- 19546107 TI - Ruptured carotid artery aneurysms of the ophthalmic (C6) segment: clinical and angiographic long term follow-up of a multidisciplinary management strategy. AB - BACKGROUND: The management of ruptured C6 aneurysms remains controversial. Detailed long-term outcome data are still lacking. Thus the present study provided a detailed long term follow-up for a multidisciplinary approach combining microsurgical clipping, endovascular embolisation and parent artery occlusion with/without bypass protection. METHODS: In our single centre analysis of 64 consecutive patients, indications for microsurgery were: superior aneurysm projection, giant/large or wide necked aneurysms and aneurysms at branching sites. Indications for embolisation were: narrow necks, neck calcification, close aneurysm relation to the clinoid process or adhesion to the distal dural ring, and aneurysm location in the concavity of the carotid siphon curve. RESULTS: 23 patients (35.9%) underwent microsurgery, 38 patients (59.4%) embolisation and three patients (4.7%) parent artery occlusion under bypass protection. Retreatment was required in 20.9% (surgery 8.7%, endovascular 31.6%). Procedure related transient complications occurred in 10.9% (surgery 13.0%, endovascular 10.5%). Procedure related permanent morbidities occurred in 6.3% (surgery 8.7%, endovascular 5.3%), including visual deficits in 4.7% (surgery 4.4%, endovascular 5.3%). One endovascular patient died. Angiographic follow-up (29.2 (SD 31.9) months) revealed total aneurysm occlusion in 94.4% of the surgical and 82.9% of the endovascular patients. Clinical follow-up (58.7 (SD 47.6) months) showed 73.4% of the population reaching Glasgow Outcome Scale 4-5, these data being equivalent to the International Subarachnoid Aneurysm Trial (ISAT) outcomes. CONCLUSIONS: Based on favourable neuroradiological and ophthalmological outcomes, microsurgery is recommended for superiorly projecting aneurysms, especially aneurysms involving the ophthalmic artery, and for giant/large or wide necked aneurysms. Based on stable aneurysm occlusion and excellent clinical outcomes, embolisation can be recommended for inferiorly/medially projecting small, narrow necked aneurysms. PMID- 19546109 TI - A reappraisal of the value of lateral spread response monitoring in the treatment of hemifacial spasm by microvascular decompression. AB - BACKGROUND: Lateral spread response (LSR) to the electrical stimulation of a facial nerve branch is a specific electrophysiological feature of primary hemifacial spasm (HFS). The curative treatment of HFS is based on surgical microvascular decompression (MVD). However, the outcome of this procedure is not always satisfactory. OBJECTIVE: To evaluate the correlation between intraoperative LSR changes and the short- and long-term postoperative clinical outcome following MVD. METHODS: Thirty-two consecutive patients with primary HFS treated by MVD performed with intraoperative LSR monitoring were retrospectively included. The patients were assessed for the presence of HFS and surgical complications at 1 day, 1 month and 6 months after surgery. The long-term clinical result was assessed between 1 and 10 years (mean 5.4 years) using a self report questionnaire. RESULTS: Patients were divided into three groups based on intraoperative LSR changes: (1) in 15 patients, LSRs were present before incision and disappeared after MVD (47%); (2) in nine patients, LSRs were present before incision but persisted despite MVD (28%); (3) in eight patients, LSRs were absent before surgery and remained so after the procedure (25%). Intraoperative LSR abolition during the MVD procedure correlated with HFS relief in the long term (p<0.0001, Fisher exact test), but not on the first day after surgery (p = 0.3564). CONCLUSIONS: Monitoring MVD by recording LSRs intraoperatively could be of value not only to indicate the resolution of the vasculonervous conflict at the end of surgery, but also to predict a successful clinical outcome in the long term after the surgical intervention. PMID- 19546116 TI - Chloroplast anchoring: its implications for the regulation of intracellular chloroplast distribution. AB - The intracellular distribution of organelles plays a pivotal role in the maintenance and adaptation of a wide spectrum of cellular activities in plants. Chloroplasts are a special type of organelle able to photosynthesize, capturing light energy to fix atmospheric CO2. Consequently, the intracellular positioning of chloroplasts is crucial for plant growth and development. Knowledge of the photoreceptors and cellular apparatus responsible for chloroplast movement has gradually accumulated over time, yet recent advances have allowed improved understanding. In this article, several aspects of research progress into the mechanisms for maintaining the specific intracellular distribution patterns of chloroplasts, namely, chloroplast anchoring, are summarized, together with a brief consideration of the future prospects of this subject. Our discussion covers developmental, physiological, ecophysiological, and recent cell biological research areas. PMID- 19546110 TI - Stochastic noise in splicing machinery. AB - The number of known alternative human isoforms has been increasing steadily with the amount of available transcription data. To date, over 100 000 isoforms have been detected in EST libraries, and at least 75% of human genes have at least one alternative isoform. In this paper, we propose that most alternative splicing events are the result of noise in the splicing process. We show that the number of isoforms and their abundance can be predicted by a simple stochastic noise model that takes into account two factors: the number of introns in a gene and the expression level of a gene. The results strongly support the hypothesis that most alternative splicing is a consequence of stochastic noise in the splicing machinery, and has no functional significance. The results are also consistent with error rates tuned to ensure that an adequate level of functional product is produced and to reduce the toxic effect of accumulation of misfolding proteins. Based on simulation of sampling of virtual cDNA libraries, we estimate that error rates range from 1 to 10% depending on the number of introns and the expression level of a gene. PMID- 19546114 TI - Development and psychometric evaluation of the reasons for living--older adults scale: a suicide risk assessment inventory. AB - PURPOSE: The purposes of these studies were to develop and initially evaluate the psychometric properties of the Reasons for Living Scale-Older Adult version (RFL OA), an older adults version of a measure designed to assess reasons for living among individuals at risk for suicide. DESIGN AND METHODS: Two studies are reported. Study 1 involved instrument development with 106 community-dwelling older adults, and initial psychometric evaluation with a second sample of 119 community-dwelling older adults. Study 2 evaluated the psychometric properties of the RFL-OA in a clinical sample. One hundred eighty-one mental health patients 50 years or older completed the RFL-OA and measures of depression, suicide ideation at the current time and at the worst point in one's life, and current mental status and physical functioning. RESULTS: Strong psychometric properties were demonstrated for the RFL-OA, with high internal consistency (Cronbach's alpha coefficient). Convergent validity was evidenced by negative associations among RFL-OA scores and measures of depression and suicide ideation. RFL-OA scores predicted current and worst-episode suicide ideation above and beyond current depression. Discriminant validity was evidenced with measures of current mental status and physical functioning. Criterion-related validity was also demonstrated with respect to lifetime history of suicidal behavior. IMPLICATIONS: These findings provide preliminary support for the validity and reliability of the RFL OA. The findings also support the potential value of attending to reasons for living during clinical treatment with depressed older adults and others at risk for suicide. PMID- 19546117 TI - A clinical prediction rule for detecting major depressive disorder in primary care: the PREDICT-NL study. AB - BACKGROUND: Major depressive disorder often remains unrecognized in primary care. OBJECTIVE: Development of a clinical prediction rule using easily obtainable predictors for major depressive disorder in primary care patients. METHODS: A total of 1046 subjects, aged 18-65 years, were included from seven large general practices in the center of The Netherlands. All subjects were recruited in the general practice waiting room, irrespective of their presenting complaint. Major depressive disorder according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Text Revision edition criteria was assessed with the Composite International Diagnostic Interview. Candidate predictors were gender, age, educational level, being single, number of presented complaints, presence of non somatic complaints, whether a diagnosis was assigned, consultation rate in past 12 months, presentation of depressive complaints or prescription of antidepressants in past 12 months, number of life events in past 6 months and any history of depression. RESULTS: The first multivariable logistic regression model including only predictors that require no confronting depression-related questions had a reasonable degree of discrimination (area under the receiver operating characteristic curve or concordance-statistic (c-statistic) = 0.71; 95% Confidence Interval (CI): 0.67-0.76). Addition of three simple though more depression-related predictors, number of life events and history of depression, significantly increased the c-statistic to 0.80 (95% CI: 0.76-0.83). After transforming this second model to an easily to use risk score, the lowest risk category (sum score < 5) showed a 1% risk of depression, which increased to 49% in the highest category (sum score > or = 30). CONCLUSION: A clinical prediction rule allows GPs to identify patients-irrespective of their complaints-in whom diagnostic workup for major depressive disorder is indicated. PMID- 19546118 TI - GP attitudes and self-reported behaviour in primary care consultations for low back pain. AB - BACKGROUND: The implementation of guideline recommendations in primary care has become widespread. The treatment of low back pain (LBP) has followed suite. Research shows that the use of LBP guidelines is influenced by the believability of the underlying evidence, the GPs consultation style and uncertainties surrounding diagnosis and treatment. OBJECTIVE: To qualitatively examine the attitudes and self-reported behaviour of GPs in relation to guideline adherence for patients with LBP. METHOD: A mixed-method design combining a national UK based survey of GPs and physiotherapists with an embedded qualitative study. This report focuses only on the GP interviews. We explored GPs' experience of managing LBP patients and the rationale for treatment offered to a patient described in a written vignette. All interviews were digitally recorded, fully transcribed and analysed using the constant comparative method. RESULTS: GPs encountered difficulties adhering to guideline recommendations for LBP patients. They experienced particular tensions between recommendations to stay active versus the expectation of being prescribed rest. GPs expressed that chronic LBP often poses an intractable problem requiring specialist advice. The perception that guideline recommendations are 'imposed' may create resistance, and the evidence base is not always considered believable. CONCLUSIONS: GPs acknowledge guideline recommendations but divergence occurs in implementation. This is due to GPs focussing on the whole person-not just one condition-and the importance of maintaining the doctor-patient relationship, which relies on effective negotiation of mutual perceptions and expectations. Further exploration on how consultation processes can be constructed to effectively combine evidence with patient-centred care is needed. PMID- 19546119 TI - Use of focus groups to develop methods to communicate cardiovascular disease risk and potential for risk reduction to people with type 2 diabetes. AB - BACKGROUND: People need to perceive a risk in order to build an intention-to change behaviour yet our ability to interpret information about risk is highly variable. OBJECTIVES: We aimed to use a user-centred design process to develop an animated interface for the UK Prospective Diabetes Study (UKPDS) Risk Engine to illustrate cardiovascular disease (CVD) risk and the potential to reduce this risk. In addition, we sought to use the same approach to develop a brief lifestyle advice intervention. METHODS: Three focus groups were held. Participants were provided with examples of materials used to communicate CVD risk and a leaflet containing a draft brief lifestyle advice intervention and considered their potential to increase motivation-to-change behaviours including diet, physical activity, and smoking in order to reduce CVD risk. Discussions were tape-recorded, transcribed and coded and recurring themes sought. RESULTS: Sixty-two percent of participants were male, mean age was 66 years (range = 47-76 years) and median age at leaving full-time education was 18 years (range = 15-40 years). Sixteen had type 2 diabetes and none had a prior history of CVD. Recurring themes from focus group discussions included the following: being less numerate is common, CVD risk reduction is important and a clear visual representation aids comprehension. CONCLUSION: A simple animated interface of the UKPDS Risk Engine to illustrate CVD risk and the potential for reducing this risk has been developed for use as a motivational tool, along with a brief lifestyle advice intervention. Future work will investigate whether use of this interactive version of the UKPDS Risk Engine and brief lifestyle advice is associated with increased behavioural intentions and changes in health behaviours designed to reduce CVD risk. PMID- 19546121 TI - Joystick interfaces are not suitable for robotized endoscope applied to NOTES. AB - BACKGROUND: NOTES has changed the working environment of endoscopy, leading to new difficulties. The limitations of conventional endoscopes call for the development of new platforms. Robotics may be the answer. MATERIALS AND METHODS: The authors compared human to robotized manipulation of a flexible endoscope into the abdominal cavity, in an animal model. Thirty-two participants were enrolled. Results were analyzed according to the clinical background of the participants: experienced endoscopists, experienced laparoscopists, and medical students. Two single-channel gastroscopes were used. Whereas one was not modified, the other had the handling wheels replaced by motors controlled through a computer and a joystick. A NOTES transgastric approach was used to access the peritoneal cavity. The time to touch previously positioned intra-abdominal numbered plastic targets was recorded 3 times with each endoscope. RESULTS: Mean time to complete the tasks was significantly shorter using the conventional endoscope (2.71 vs 6.96 minutes, P < .001). When the robotized endoscope was used, the mean times of endoscopists (7.42 minutes), laparoscopists (6.84 minutes), and students (6.77 minutes) were statistically identical. No differences were found between laparoscopists and students in both techniques. DISCUSSION: Applying robotics to a flexible endoscope fails to enhance ability to move into the abdominal cavity, partly because of the interface. To overcome the limitations of endoscope when performing complex NOTES tasks, robotics may be useful, especially to control the instruments and to stabilize the endoscope itself. CONCLUSION: Robotized endoscope with joystick interface is not sufficient to enhance immediate intuitiveness of flexible endoscopy applied to NOTES. PMID- 19546122 TI - Anal incontinence improvement after silicone injection may be related to restoration of sphincter asymmetry. AB - BACKGROUND: This study aimed to evaluate manometric parameters that may explain improvement in anal incontinence using a silicone bulking agent. METHODS: Incontinent patients having internal sphincter defects were prospectively selected and injected with a silicone bulking agent. Manometry and endoanal ultrasound were performed before and 3 months after injections. Twenty continent healthy volunteers were used only for manometric comparison. RESULTS: Thirty-five patients (28 females; mean age 60.3 years) and 20 controls entered this study. Patients had lower resting and squeeze pressures compared with controls (P < .05). Length of the high-pressure zone increased from 1 to 1.7 cm postinjection (P = .002). Asymmetry index showed a significant change postinjection (P < .001). CONCLUSION: Despite considerable clinical improvement, no significant increase in manometric pressures was noted posttreatment. There was significant improvement in both high-pressure zone and asymmetry index, and these findings may explain the mechanism of action of the bulking agent injected. PMID- 19546120 TI - Chromosomal mapping of canine-derived BAC clones to the red fox and American mink genomes. AB - High-quality sequencing of the dog (Canis lupus familiaris) genome has enabled enormous progress in genetic mapping of canine phenotypic variation. The red fox (Vulpes vulpes), another canid species, also exhibits a wide range of variation in coat color, morphology, and behavior. Although the fox genome has not yet been sequenced, canine genomic resources have been used to construct a meiotic linkage map of the red fox genome and begin genetic mapping in foxes. However, a more detailed gene-specific comparative map between the dog and fox genomes is required to establish gene order within homologous regions of dog and fox chromosomes and to refine breakpoints between homologous chromosomes of the 2 species. In the current study, we tested whether canine-derived gene-containing bacterial artificial chromosome (BAC) clones can be routinely used to build a gene-specific map of the red fox genome. Forty canine BAC clones were mapped to the red fox genome by fluorescence in situ hybridization (FISH). Each clone was uniquely assigned to a single fox chromosome, and the locations of 38 clones agreed with cytogenetic predictions. These results clearly demonstrate the utility of FISH mapping for construction of a whole-genome gene-specific map of the red fox. The further possibility of using canine BAC clones to map genes in the American mink (Mustela vison) genome was also explored. Much lower success was obtained for this more distantly related farm-bred species, although a few BAC clones were mapped to the predicted chromosomal locations. PMID- 19546123 TI - The use of holmium laser technology for the treatment of refractory common bile duct stones, with a short review of the relevant literature. AB - BACKGROUND: The treatment of common bile duct (CBD) stones can vary in complexity and many methods exist to fragment them before removal. Although holmium laser is frequently used in urological surgery, it is rarely used to achieve this aim. METHODS: The holmium laser was passed along a fiber introduced via a flexible scope through the cystic duct at the time of laparoscopic cholecystectomy. This energy modality was used to fragment the stones to a size that allowed easy removal. RESULTS: The authors have used this technique once so far and achieved complete clearance of the CBD with no mucosal damage. CONCLUSION: Holmium laser provides an alternative and realistic treatment option for difficult CBD stones. PMID- 19546124 TI - New hybrid approach for NOTES transvaginal cholecystectomy: preliminary clinical experience. AB - OBJECTIVES: Natural orifice translumenal endoscopic surgery (NOTES) represents the first step toward scar-less surgery. The objective of this study is to evaluate early clinical results of transvaginal cholecystectomy using a new technique. METHODS: Institutional review board approval was obtained and transvaginal NOTES cholecystectomy was performed in 12 women for cholelithiasis. A 2-channel videoendoscope was inserted in the abdominal cavity through a posterior colpotomy. Two 3-mm trocars were inserted deep in the umbilicus, and a 10-mm trocar was placed through the colpotomy parallel to the endoscope. Dissection was performed with endoscopic instruments combined with 3-mm laparoscopic instruments. RESULTS: Mean operative time was 125.8 minutes. All procedures occurred without intraoperative complications or conversions, except for 1 vulvar laceration. There were no postoperative complications in the clinical follow-up. CONCLUSION: Transvaginal NOTES is a feasible and safe alternative for cholecystectomy in this preliminary clinical experience, allowing good cosmetic benefits and low analgesic requirement. PMID- 19546125 TI - Training and working in high-stakes environments: lessons learned and problems shared by aviators and surgeons. AB - Surgeons and naval aviators are both trained to work in high-stakes environments. Any misadventure in either of their working worlds can lead to death. Yet the pathways to certification and implicit attitudes toward training are quite different in these 2 disciplines and provide an opportunity to compare and contrast the methodologies employed. At the 5th annual Innovations in the Surgical Environments Conference, senior and junior aviators and surgeons shared their experiences from the perspective of trainee and trainer and in the process presented an interesting study in parallels and contrasts. The US Navy follows a highly regimented training syllabus with graduated levels of responsibility designed to create the safest possible flying environment. Extensive preflight and postflight effort is required for each mission flown. Surgical training is also hierarchal in responsibility, but graduates demonstrate greater variability in their training experience. The surgical field can only fortify its emphasis on safety by seeking to provide the optimal training experiences necessary in the high-stakes environment of the operating theater. In doing so, surgeons may find reinvigorated commitment through study of the aviation industry's established methods of training and practice. PMID- 19546126 TI - Occupational cancer: an emerging problem in newly industrializing countries. PMID- 19546127 TI - Prevalence of work-related upper limbs symptoms (WRULS) among office workers. AB - An analytical cross-sectional study was carried out to determine the prevalence of work-related upper limbs symptoms (WRULS) among office workers and factors associated with it. A self-administered questionnaire was used to collect the information. A total of 463 (87.7%) office workers from selected government agencies participated in this study. The mean age of the respondents was 34.1 years (range = 18-55 years). Majority (91.6%) were Malay and female (72.8%), and 58.1% were from clerical group. Mean duration of work was 8.7 years. This study found that prevalence of WRULS was 33.0% (95% CI [confidence interval] = 28.8%, 37.3%). Computer users at work had 2.0 (95% CI = 1.1, 3.4) higher odds ratio of developing WRULS and those who used it for 5 hours and more per day had 7.5 (95% CI = 2.3, 24.2) higher odds ratio of developing WRULS. Hand-intensive hobbies and higher education were also found to be associated with WRULS. PMID- 19546128 TI - The transformation of vascular surgeons to vascular specialists: policy or necessity? AB - The wide spreading of vascular diseases along with the emergence of minimally invasive endovascular therapies and modern medical therapies is inevitably bringing many disciplines into play. Although by definition, vascular surgery is the clinical and scientific discipline concerned with the diagnosis, treatment, and prevention of vascular diseases, many more radiologists and a significant number of cardiologists become increasingly involved. The question of which specialist among those qualified should carry out medical treatment, open, and endovascular procedures is still suspended, and a professional competition is pronounced. This article discusses the necessity of a vascular specialist who guides the holistic management of vascular diseases: open surgery, endovascular intervention, and medical therapy. The different characteristics of those intending to offer vascular care, the training curricula, the workforce demands, and the role of vascular centers are analyzed to illustrate the keystone role of vascular surgeons who are currently transforming to vascular specialists. PMID- 19546129 TI - An aneurysm of the external iliac vein. PMID- 19546130 TI - Genetics and genomics in nursing research. PMID- 19546132 TI - US is now "past the rhetoric" in fighting malaria worldwide. PMID- 19546133 TI - German agency refuses to rule on drug's benefits until Pfizer discloses all trial results. PMID- 19546134 TI - Smoking in pregnancy. PMID- 19546135 TI - The extremely premature neonate: anticipating and managing care. PMID- 19546136 TI - Vaccine disputes. PMID- 19546137 TI - Long term exposure to air pollution decreases life expectancy, UK report finds. PMID- 19546138 TI - US Congress tells FDA to regulate tobacco. PMID- 19546140 TI - Obama asks AMA to support his healthcare reform package. PMID- 19546142 TI - Would that we had the energy. PMID- 19546143 TI - Reimbursement for drugs -- a register study comparing economic outcome for five healthcare centres in areas with different socioeconomic conditions. AB - AIMS: Previous studies have indicated the negative effects of socioeconomic deprivation on health status and morbidity. Nevertheless, the economic assignment systems for pharmaceutical benefits in Sweden do not take socioeconomic status (SES) into account. The aim of the study was, therefore, to compare reimbursement for subsidized drugs at primary healthcare centres (HCCs) with differing socioeconomic conditions in relation to real costs. The word reimbursement is used to denote economic compensation to the HCCs from the county council for drug benefit costs. METHODS: The numbers of individuals dispensed drugs, total costs and reimbursement at five HCCs with different socioeconomic conditions were compared. A socioeconomic index was calculated for each HCC on the basis of information from the municipality registries on income (with negative sign), assistance allowance, education, foreign background, and unemployment. Register data on drug benefit costs were retrieved from the National Corporation of Pharmacies (Apoteket AB) and the Swedish Prescribed Drug Register at the National Board of Health and Welfare. Data on listed and unlisted citizens at the Kalmar County Council and on public statistics from registers at the HCC municipalities where the HCCs were situated were retrieved. RESULTS: There was an almost inverse linear relationship between total cost compensation and the socioeconomic index (n = 5; r =-0.99; p = 0.001). The HCCs with the lowest SES received lower cost compensation. CONCLUSIONS: HCCs responsible for citizens with lower SES appeared to be disadvantaged by the prevalent reimbursement system in Sweden, thereby increasing differences in the state of health of the citizens. This, in turn, hampers health preventing programmes and lifestyle interventions. An HCC-specific standardized summary of socioeconomic burden is presented. PMID- 19546144 TI - STEM imaging of 47-pm-separated atomic columns by a spherical aberration corrected electron microscope with a 300-kV cold field emission gun. AB - A spherical aberration-corrected electron microscope has been developed recently, which is equipped with a 300-kV cold field emission gun and an objective lens of a small chromatic aberration coefficient. A dumbbell image of 47 pm spacing, corresponding to a pair of atomic columns of germanium aligned along the [114] direction, is resolved in high-angle annular dark field (HAADF) scanning transmission electron microscopy (STEM) with a 0.4-eV energy spread of the electron beam. The observed image was compared with a simulated image obtained by dynamical calculation. PMID- 19546145 TI - Foucauldian diagnostics: space, time, and the metaphysics of medicine. AB - This essay places Foucault's work into a philosophical context, recognizing that Foucault is difficult to place and demonstrates that Foucault remains in the Kantian tradition of philosophy, even if he sits at the margins of that tradition. For Kant, the forms of intuition-space and time-are the a priori conditions of the possibility of human experience and knowledge. For Foucault, the a priori conditions are political space and historical time. Foucault sees political space as central to understanding both the subject and objects of medicine, psychiatry, and the social sciences. Through this analysis one can see that medicine's metaphysics is a metaphysics of efficient causation, where medicine's objects are subjected to mechanisms of efficient control. PMID- 19546146 TI - Governmentality, biopower, and the debate over genetic enhancement. AB - Although Foucault adamantly refused to make moral pronouncements or dictate moral principles or political programs to his readers, his work offers a number of tools and concepts that can help us develop our own ethical views and practices. One of these tools is genealogical analysis, and one of these concepts is "biopower." Specifically, this essay seeks to demonstrate that Foucault's concept of biopower and his genealogical method are valuable as we consider moral questions raised by genetic enhancement technologies. First, it examines contemporary debate over the development, marketing, and application of such technologies, suggesting that what passes for ethical deliberation is often little more than political maneuvering in a field where stakes are very high and public perceptions will play a crucial role in decisions about which technologies will be funded or disallowed. It goes on to argue that genuine ethical deliberation on these issues requires some serious investigation of their historical context. Accordingly, then, it takes up the oft-heard charge from critics that genetic enhancement technologies are continuous with twentieth century eugenic projects or will usher in a new age of eugenics. Foucault explicitly links twentieth-century eugenics with the rise of biopower. Through review of some aspects of the twentieth-century eugenics movement alongside some of the rhetoric and claims of enhancement's modern-day proponents, the essay shows ways in which deployment of genetic enhancement technologies is and is not continuous with earlier deployments of biopower. PMID- 19546147 TI - Foucault, genealogy, ethics. PMID- 19546148 TI - Physiological and molecular approaches to improve drought resistance in soybean. AB - Drought stress is a major constraint to the production and yield stability of soybean [Glycine max (L.) Merr.]. For developing high yielding varieties under drought conditions, the most widely employed criterion has traditionally been direct selection for yield stability over multiple locations. However, this approach is time consuming and labor intensive, because yield is a highly quantitative trait with low heritability, and influenced by differences arising from soil heterogeneity and environmental factors. The alternative strategy of indirect selection using secondary traits has succeeded only in a few crops, due to problems with repeatability and lack of phenotyping strategies, especially for root-related traits. Considerable efforts have been directed towards identifying traits associated with drought resistance in soybean. With the availability of the whole genome sequence, physical maps, genetics and functional genomics tools, integrated approaches using molecular breeding and genetic engineering offer new opportunities for improving drought resistance in soybean. Genetic engineering for drought resistance with candidate genes has been reported in the major food crops, and efforts for developing drought-resistant soybean lines are in progress. The objective of this review is to consolidate the current knowledge of physiology, molecular breeding and functional genomics which may be influential in integrating breeding and genetic engineering approaches for drought resistance in soybean. PMID- 19546149 TI - Evaluation of the Wound Healing Potential of Achillea biebersteinii Afan. (Asteraceae) by In Vivo Excision and Incision Models. AB - Achillea species are widely used for diarrhea, abdominal pain, stomachache and healing of wounds in folk medicine. To evaluate the wound healing activity of the plant, extracts were prepared with different solvents; hexane, chloroform, ethyl acetate and methanol, respectively from the roots of Achillea biebersteinii. Linear incision by using tensiometer and circular excision wound models were employed on mice and rats. The wound healing effect was comparatively evaluated with the standard skin ointment Madecassol. The n-hexane extract treated groups of animals showed 84.2% contraction, which was close to contraction value of the reference drug Madecassol (100%). On the other hand the same extract on incision wound model demonstrated a significant increase (40.1%) in wound tensile strength as compared to other groups. The results of histoptological examination supported the outcome of linear incision and circular excision wound models as well. The experimental data demonstrated that A. biebersteinii displayed remarkable wound healing activity. PMID- 19546150 TI - Cognitive function and overweight in preschool children. AB - The authors assessed the association between cognitive function and incidence and maintenance of overweight in preschool children. A population-based birth cohort was established in Menorca, Spain, between 1997 and 1999 (n = 482). Body mass index (weight (kg)/height (m)(2)) was measured at ages 4 years and 6 years (n = 421). At age 4 years, children were assessed for cognitive function (McCarthy Scales of Children's Abilities) (n = 395). After multivariable adjustment for a wide range of factors, including maternal education and body mass index, children with higher general cognition at age 4 years had a lower likelihood of being overweight (odds ratio = 0.47, 95% confidence interval (CI): 0.25, 0.88) at age 6 years. Children with higher general cognition at age 4 years had a lower likelihood of maintaining an unhealthy weight status (being at risk of overweight or overweight) between ages 4 years and 6 years, as well as worsening weight status over time, than children who maintained a healthy weight (odds ratios were 0.78 (95% CI: 0.54, 1.14) and 0.77 (95% CI: 0.51, 1.14), respectively). When specific dimensions of cognitive function were assessed, associations were mainly found for verbal and executive function areas. Children with higher cognitive function in early life might be at decreased risk of overweight later in childhood. PMID- 19546151 TI - Age of onset in concordant twins and other relative pairs with multiple sclerosis. AB - The ages of onset in multiple sclerosis cases span more than 7 decades. Data are presented for affected relative pairs from a Canadian population base of 30,000 multiple sclerosis index cases (1993-2008). The effects of genetic sharing, parent of origin, intergenerational versus collinear differences, and gender on the ages of onset were evaluated in the following concordant pairs: monozygotic twins (n = 29), dizygotic twins (n = 10), siblings (n = 614), first cousins (n = 405), half siblings (n = 29), parent/child (n = 285), and aunt/uncle/niece/nephew (avunculars) (n = 289). Fisher's z test assessed intraclass correlation (r) for ages of onset. Correlations for monozygotic twins, dizygotic twins, full siblings, and first cousins were 0.60, 0.54, 0.20, and 0.10, respectively. Dizygotic twins resembled monozygotic twins more than siblings. The age-of-onset correlation for maternal half siblings (r = 0.37) was higher than that for paternal half siblings (r = 0.26), consistent with other observations suggesting an intrauterine environmental effect on multiple sclerosis risk. Intergenerational comparisons are complicated by substantial increases of multiple sclerosis incidence over time. Genetic loading (familial vs. sporadic cases) did not generally influence the age of onset, but correlation of age of onset in multiple sclerosis relative pairs was proportional to genetic sharing. A maternal parent-of-origin effect on the age of onset in collinear generations was suggested. PMID- 19546152 TI - Hepatitis C infection among injecting drug users in England and Wales (1992 2006): there and back again? AB - Changes in hepatitis C virus (HCV) prevalence from 1992 to 2006 were examined by using 24,311 records from unlinked anonymous surveillance of injecting drug users in England and Wales. Bayesian logistic regression was used to estimate annual prevalence, accounting for changing recruitment patterns (age, gender, injecting duration, geographic region, interactions) and the sensitivity and specificity of different oral fluid testing devices. After controlling for these differences, the authors found that the adjusted HCV prevalence decreased from 70% (95% credible interval: 62, 78) in 1992 to 47% (95% credible interval: 43, 51) in 1998 before rising again to 53% (95% credible interval: 48, 58) in 2006. Women injecting drug users had a higher HCV risk than did men (odds ratio = 1.50, 95% credible interval: 1.31, 1.73). Two regions (London and North West) had a markedly higher HCV prevalence than did the rest of England and Wales. Among individuals who had injected for less than 1 year, the adjusted HCV prevalence in 2006 was higher than that in 1992 (28% vs. 19%, respectively). HCV infection can be prevented. The public health challenge in England and Wales is to increase action in order to regain a downward trend in HCV risk and the benefit that has been lost since 1998. PMID- 19546153 TI - Ideal weight and weight satisfaction: association with health practices. AB - Evidence suggests that individuals have become more tolerant of higher body weights over time. To investigate this issue further, the authors examined cross sectional associations among ideal weight, examination year, and obesity as well as the association of ideal weight and body weight satisfaction with health practices among 15,221 men and 4,126 women in the United States. Participants in 1987 reported higher ideal weights than participants in 2001, an effect particularly pronounced from 1987 to 2001 for younger and obese men (85.5 kg to 94.9 kg) and women (62.2 kg to 70.5 kg). For a given body mass index, higher ideal body weights were associated with greater weight satisfaction but lower intentions to lose weight. Body weight satisfaction was subsequently associated with greater walking/jogging, better diet, and lower lifetime weight loss but with less intention to change physical activity and diet or lose weight (P < 0.01). Conversely, body mass index was negatively associated with weight satisfaction (P < 0.01) and was associated with less walking/jogging, poorer diet, and greater lifetime weight loss but with greater intention to change physical activity and diet or lose weight. Although the health implications of these findings are somewhat unclear, increased weight satisfaction, in conjunction with increases in societal overweight/obesity, may result in decreased motivation to lose weight and/or adopt healthier lifestyle behaviors. PMID- 19546155 TI - Topographical functional connectivity pattern in the perisylvian language networks. AB - We performed a resting-state functional connectivity study to investigate directly the functional correlations within the perisylvian language networks by seeding from 3 subregions of Broca's complex (pars opercularis, pars triangularis, and pars orbitalis) and their right hemisphere homologues. A clear topographical functional connectivity pattern in the left middle frontal, parietal, and temporal areas was revealed for the 3 left seeds. This is the first demonstration that a functional connectivity topology can be observed in the perisylvian language networks. The results support the assumption of the functional division for phonology, syntax, and semantics of Broca's complex as proposed by the memory, unification, and control (MUC) model and indicated a topographical functional organization in the perisylvian language networks, which suggests a possible division of labor for phonological, syntactic, and semantic function in the left frontal, parietal, and temporal areas. PMID- 19546154 TI - Activation of sensory-motor areas in sentence comprehension. AB - The sensory-motor account of conceptual processing suggests that modality specific attributes play a central role in the organization of object and action knowledge in the brain. An opposing view emphasizes the abstract, amodal, and symbolic character of concepts, which are thought to be represented outside the brain's sensory-motor systems. We conducted a functional magnetic resonance imaging study in which the participants listened to sentences describing hand/arm action events, visual events, or abstract behaviors. In comparison to visual and abstract sentences, areas associated with planning and control of hand movements, motion perception, and vision were activated when understanding sentences describing actions. Sensory-motor areas were activated to a greater extent also for sentences with actions that relied mostly on hands, as opposed to arms. Visual sentences activated a small area in the secondary visual cortex, whereas abstract sentences activated superior temporal and inferior frontal regions. The results support the view that linguistic understanding of actions partly involves imagery or simulation of actions, and relies on some of the same neural substrate used for planning, performing, and perceiving actions. PMID- 19546156 TI - Altered water diffusivity in cortical association tracts in children with early deprivation identified with Tract-Based Spatial Statistics (TBSS). AB - Institutional rearing is associated with neurocognitive and behavioral difficulties. Although such difficulties are thought to reflect abnormal neurologic development resulting from early social deprivation (ED) and there is evidence for functional abnormality in children with histories of ED, the impact of early deprivation on brain anatomy has received little study in humans. The present study utilized an objective and sensitive neuroimaging analysis technique (Tract-Based Spatial Statistics) to evaluate white matter fractional anisotropy (FA) and diffusivity in a group of right-handed children with histories of ED (n = 17; mean age = 10.9 + 2.6 years) as compared with age-matched healthy controls (n = 15; mean age = 11.7 + or - 2.8 years). Participants underwent magnetic resonance imaging diffusion tensor imaging sequences and comprehensive neuropsychological evaluations. Results revealed reduced FA in frontal, temporal, and parietal white matter including components of uncinate and superior longitudinal fasciculi, in children with histories of ED, providing further support for limbic and paralimbic abnormalities in children with such histories. Furthermore, white matter abnormalities were associated with duration of time in the orphanage and with inattention and hyperactivity scores. It is suspected that the observed white matter abnormalities are associated with multiple depriving factors (e.g., poor prenatal care, postnatal stress) associated with institutional caregiving. PMID- 19546157 TI - Cerebellar engagement in an action observation network. AB - The cerebellum has traditionally been viewed as a brain structure subserving skilled motor behaviors. However, the cerebellum might be involved not only in movement coordination, but also in action observation and understanding of others' actions. Veridical visual perception of human body motion is of immense importance for a variety of daily-life situations and for successful social interactions. Here, by combining visual psychophysics with a lesion analysis, we assessed visual sensitivity to human walking in patients with lesions to the left cerebellum. Patients with left lateral cerebellar lesions exhibit deficits in visual sensitivity to body motion, whereas medial lesions do not substantially affect visual perception of human locomotion. The findings point to left lateral cerebellar involvement in an action observation network. We discuss possible mechanisms of cerebellar engagement in visual social perception revealed by body motion. PMID- 19546158 TI - Protease gene duplication and proteolytic activity in Drosophila female reproductive tracts. AB - Secreted proteases play integral roles in sexual reproduction in a broad range of taxa. In the genetic model Drosophila melanogaster, these molecules are thought to process peptides and activate enzymes inside female reproductive tracts, mediating critical postmating responses. A recent study of female reproductive tract proteins in the cactophilic fruit fly Drosophila arizonae, identified pervasive, lineage-specific gene duplication amongst secreted proteases. Here, we compare the evolutionary dynamics, biochemical nature, and physiological significance of secreted female reproductive serine endoproteases between D. arizonae and its congener D. melanogaster. We show that D. arizonae lower female reproductive tract (LFRT) proteins are significantly enriched for recently duplicated secreted proteases, particularly serine endoproteases, relative to D. melanogaster. Isolated lumen from D. arizonae LFRTs, furthermore, exhibits significant trypsin-like and elastase-like serine endoprotease activity, whereas no such activity is seen in D. melanogaster. Finally, trypsin- and elastase-like activity in D. arizonae female reproductive tracts is negatively regulated by mating. We propose that the intense proteolytic environment of the D. arizonae female reproductive tract relates to the extraordinary reproductive physiology of this species and that ongoing gene duplication amongst these proteases is an evolutionary consequence of sexual conflict. PMID- 19546159 TI - Acrylonitrile-induced oxidative stress and oxidative DNA damage in male Sprague Dawley rats. AB - Studies have demonstrated that the induction of oxidative stress may be involved in brain tumor induction in rats by acrylonitrile. The present study examined whether acrylonitrile induces oxidative stress and DNA damage in rats and whether blood can serve as a valid surrogate for the biomonitoring of oxidative stress induced by acrylonitrile in the exposed population. Male Sprague-Dawley rats were treated with 0, 3, 30, 100, and 200 ppm acrylonitrile in drinking water for 28 days. One group of rats were also coadministered N-acetyl cysteine (NAC) (0.3% in diet) with acrylonitrile (200 ppm in drinking water) to examine whether antioxidant supplementation was protective against acrylonitrile-induced oxidative stress. Direct DNA strand breakage in white blood cells (WBC) and brain was measured using the alkaline comet assay. Oxidative DNA damage in WBC and brain was evaluated using formamidopyrimidine DNA glycosylase (fpg)-modified comet assay and with high-performance liquid chromatography-electrochemical detection. No significant increase in direct DNA strand breaks was observed in brain and WBC from acrylonitrile-treated rats. However, oxidative DNA damage (fpg comet and 8'hydroxyl-2-deoxyguanosine) in brain and WBC was increased in a dose dependent manner. In addition, plasma levels of reactive oxygen species (ROS) increased in rats administered acrylonitrile. Dietary supplementation with NAC prevented acrylonitrile-induced oxidative DNA damage in brain and WBC. A slight, but significant, decrease in the GSH:GSSG ratio was seen in brain at acrylonitrile doses > 30 ppm. These results provide additional support that the mode of action for acrylonitrile-induced astrocytomas involves the induction of oxidative stress and damage. Significant associations were seen between oxidative DNA damage in WBC and brain, ROS formation in plasma, and the reported tumor incidences. Since oxidative DNA damage in brain correlated with oxidative damage in WBC, these results suggest that monitoring WBC DNA damage maybe a useful tool to assess acrylonitrile-induced oxidative stress in humans. PMID- 19546160 TI - Met receptor tyrosine kinase transactivation is involved in proteinase-activated receptor-2-mediated hepatocellular carcinoma cell invasion. AB - The expression of proteinase-activated receptor (PAR)(2) in human hepatocellular carcinoma (HCC) was established by reverse transcription-polymerase chain reaction, confocal immunofluorescence and electron microscopy in permanent cell lines, primary HCC cell cultures and HCC tumor tissue. Stimulation of HCC cells with trypsin and the PAR(2)-selective activating peptide, 2-furoyl-LIGRLO-NH(2), increased cell invasion across Matrigel. Both effects were blocked by a PAR(2) selective pepducin antagonist peptide (pal-PAR(2)) and by PAR(2) silencing with specific small interfering RNA (siRNA). PAR(2)-initiated HCC cell invasion was also blocked by inhibiting the hepatocyte growth factor receptor (Met receptor tyrosine kinase) with the receptor-targeted kinase inhibitors, SU 11274 and PHA 665752, or by downregulation of Met with specific siRNA. The involvement of Met in PAR(2)-mediated HCC invasive signaling was further supported by the finding that treatment of HCC cells with trypsin or the PAR(2)-selective agonist peptide, 2-furoyl-LIGRLO-NH(2), stimulated Met activation-phosphorylation. In addition, Met-dependent stimulation of p42/p44 mitogen-activated protein Kinases was found to be critical for the PAR(2)-Met receptor tyrosine kinase-invasive signaling axis in HCC cells. Our study establishes an important link between the PAR(2) and Met receptor tyrosine kinase signaling in promoting HCC cell invasion. PMID- 19546161 TI - Dual role of Ski in pancreatic cancer cells: tumor-promoting versus metastasis suppressive function. AB - Ski used to be defined as an oncogene that contributes to the resistance of tumor cells to transforming growth factor-beta (TGF-beta)-induced growth arrest. As TGF beta has a dual effect on tumor growth with both tumor-suppressing and -promoting activity depending on the stage of carcinogenesis and the cell type, the precise role of Ski in carcinogenesis remains unclear. In this study, we show that downregulation of Ski through lentivirus-mediated RNA interference decreases tumor growth both in vitro and in vivo, yet promotes cell invasiveness in vitro, and lung metastasis in vivo in the pancreatic cancer cell line SW1990, which contain wild-type Smad4 expression, and the BxPC3 cell line, which is Smad4 deficient. We also show that the downregulation of Ski increases TGF-beta-induced transcriptional activity, which is associated with increased TGF-beta-dependent Smad2/3 phosphorylation, and results in an altered expression profile of TGF-beta inducible genes involved in metastasis, angiogenesis and cell proliferation and epithelial-mesenchymal transition. Immunohistochemical analysis of specimens from 71 patients with pancreatic adenocarcinoma showed a significant association between overexpression of Ski and decreased patient survival time (P = 0.0024). Our results suggest that Ski may act as a tumor proliferation-promoting factor or as a metastatic suppressor in human pancreatic cancer. PMID- 19546162 TI - Real-time monitoring of cAMP levels in living endothelial cells: thrombin transiently inhibits adenylyl cyclase 6. AB - The crosstalk between Ca(2+) and cAMP signals plays a significant role for the regulation of the endothelial barrier function. The Ca(2+)-elevating agent thrombin was demonstrated to increase endothelial permeability and to decrease cAMP levels. Since Ca(2+) and cAMP signals are highly dynamic, we aimed to study the temporal resolution between thrombin-evoked Ca(2+) signals and subsequent changes of cAMP levels. Here we conduct the first real-time monitoring of thrombin-mediated regulation of cAMP signals in intact human umbilical vein endothelial cells (HUVECs) by utilising the Ca(2+)-sensitive dye Fluo-4 and the fluorescence resonance energy transfer (FRET)-based cAMP sensor Epac1-camps. We calibrated in vitro FRET responses of Epac1-camps to [cAMP] in order to estimate changes in intracellular [cAMP] evoked by thrombin treatment of HUVECs. After increasing [cAMP] to 1.2 +/- 0.2 microm by stimulation of HUVECs with isoproterenol (isoprenaline), we observed a transient decrease of cAMP levels by 0.4 +/- 0.1 microm which reached a minimum value 30 s after thrombin application and 15 s after the thrombin-evoked Ca(2+) peak. This transient decrease in [cAMP] was Ca(2+)-dependent and independent of a G(i)-mediated inhibition of adenylyl cyclases (ACs). Instead the knock down of the predominant subtype AC6 in HUVECs provided the first direct evidence that the Ca(2+)-mediated inhibition of AC6 accounts for the thrombin-induced decrease in cAMP levels. PMID- 19546163 TI - Electrophysiological properties of thalamic, subthalamic and nigral neurons during the anti-parkinsonian placebo response. AB - Placebo administration to Parkinson patients is known to induce dopamine release in the striatum and to affect the activity of subthalamic nucleus (STN) neurons. By using intraoperative single-neuron recording techniques in awake patients, here we extend our previous study on STN recording, and characterize part of the neuronal circuit which is affected by placebos. In those patients who showed a clinical placebo response, there was a decrease in firing rate in STN neurons that was associated with a decrease in the substantia nigra pars reticulata (SNr) and an increase in the ventral anterior (VA) and anterior ventral lateral (VLa) thalamus. These data show that placebo decreases STN and SNr activity whereas it increases VA/VLa activity. By contrast, placebo non-responders showed either a lack of changes in this circuit or partial changes in the STN only. Thus, changes in activity in the whole basal ganglia-VA/VLa circuit appear to be important in order to observe a clinical placebo improvement, although the involvement of other circuits, such as the direct pathway bypassing the STN, cannot be ruled out. The circuit we describe in the present study is likely to be a part of a more complex circuitry, including the striatum and the internal globus pallidus (GPi), that is modified by placebo administration. These findings indicate that a placebo treatment, which is basically characterized by verbal suggestions of benefit, can reverse the malfunction of a complex neuronal circuit, although these placebo-associated neuronal changes are short-lasting and occur only in some patients but not in others. PMID- 19546164 TI - Angiomotin p80/p130 ratio: a new indicator of exercise-induced angiogenic activity in skeletal muscles from obese and non-obese rats? AB - Skeletal muscle capillarisation responds to physiological and pathological conditions with a remarkable plasticity. Angiomotin was recently identified as a new pro-angiogenic molecule. Angiomotin is expressed as two protein isoforms, p80 and p130. Whereas p80 stimulates endothelial cell migration and angiogenesis, p130 is rather characteristic of stabilized and matured vessels. To date, how angiomotin expression is physiologically regulated in vivo remains largely unknown. We thus investigated (1) whether angiomotin was physiologically expressed in skeletal muscle; (2) whether exercise training, known to stimulate muscle angiogenesis, affected angiomotin expression; and (3) whether such regulation was altered in obesity, a pathological situation often associated with an impaired angiogenic activity and some capillary rarefaction in skeletal muscle. Two models of obesity were used: a high fat diet regime and Zucker Diabetic Fatty rats (ZDF). Our results provide evidence that angiomotin was expressed both in capillaries and myofibres. In non-obese rats, the p80 isoform was increased in plantaris muscle in response to endurance training whereas p130 was unaffected. In obese animals, no change was observed for p80 whereas training significantly decreased p130 expression. Exercise training induced angiogenesis in plantaris from both obese and non-obese rats, possibly through the modulation of angiomotin level and its consequences on RhoA-ROCK signalling. In conclusion, any increase in p80 or decrease in p130, as respectively observed in non-obese and obese animals, led to an increased ratio between p80 and p130 isoforms. This increased angiomotin p80/p130 ratio might then directly reflect the enhanced angiogenic ability of skeletal muscle in response to exercise training. PMID- 19546166 TI - A semiparametric 2-part mixed-effects heteroscedastic transformation model for correlated right-skewed semicontinuous data. AB - In longitudinal or hierarchical structure studies, we often encounter a semicontinuous variable that has a certain proportion of a single value and a continuous and skewed distribution among the rest of values. In this paper, we propose a new semiparametric 2-part mixed-effects transformation model to fit correlated skewed semicontinuous data. In our model, we allow the transformation to be nonparametric. Fitting the proposed model faces computational challenges due to intractable numerical integrations. We derive the estimates for the parameter and the transformation function based on an approximate likelihood, which has high-order accuracy but less computational burden. We also propose an estimator for the expected value of the semicontinuous outcome on the original scale. Finally, we apply the proposed methods to a clinical study on effectiveness of a collaborative care treatment on late-life depression on health care costs. PMID- 19546165 TI - Calcium diffusion enhanced after cleavage of negatively charged components of brain extracellular matrix by chondroitinase ABC. AB - The concentration of extracellular calcium plays a critical role in synaptic transmission and neuronal excitability as well as other physiological processes. The time course and extent of local fluctuations in the concentration of this ion largely depend on its effective diffusion coefficient (D*) and it has been speculated that fixed negative charges on chondroitin sulphate proteoglycans (CSPGs) and other components of the extracellular matrix may influence calcium diffusion because it is a divalent cation. In this study we used ion-selective microelectrodes combined with pressure ejection or iontophoresis of ions from a micropipette to quantify diffusion characteristics of neocortex and hippocampus in rat brain slices. We show that D* for calcium is less than the value predicted from the behaviour of the monovalent cation tetramethylammonium (TMA), a commonly used diffusion probe, but D* for calcium increases in both brain regions after the slices are treated with chondroitinase ABC, an enzyme that predominantly cleaves chondroitin sulphate glycans. These results suggest that CSPGs do play a role in determining the local diffusion properties of calcium in brain tissue, most likely through electrostatic interactions mediating rapid equilibrium binding. In contrast, chondroitinase ABC does not affect either the TMA diffusion or the extracellular volume fraction, indicating that the enzyme does not alter the structure of the extracellular space and that the diffusion of small monovalent cations is not affected by CSPGs in the normal brain ionic milieu. Both calcium and CSPGs are known to have many distinct roles in brain physiology, including brain repair, and our study suggests they may be functionally coupled through calcium diffusion properties. PMID- 19546167 TI - Molecular characterization of novel germline deletions affecting SDHD and SDHC in pheochromocytoma and paraganglioma patients. AB - A major cause of paraganglioma and pheochromocytoma is germline mutation of the tumor suppressor genes SDHB, SDHC, and SDHD, encoding subunits of succinate dehydrogenase (SDH). While many SDH missense/nonsense mutations have been identified, few large deletions have been described. We performed multiplex ligation-dependent probe amplification deletion analysis in 126 point mutation negative patients, and here we describe four novel deletions of SDHD and SDHC. Long-range PCR was used for the fine mapping of deletions. One patient had a 10 kb AluSg-AluSx-mediated deletion including SDHD exons 1 and 2, the entire TIMM8B gene, and deletion of exons of C11orf57. A second patient had a deletion of SDHD exons 1 and 2 and exon 1 of the TIMM8B gene. A third patient showed a deletion of exon 2 of SDHD, together with a 235 bp MIRb-Tensin gene insertion. In a fourth patient, a deletion of exons 5 and 6 of the SDHC gene was found, only the second SDHC deletion currently known. The deletions of the TIMM8B and C11orf57 genes are the first to be described, but do not appear to result in an additional phenotype in these patients. Four of the eight breakpoints occurred in Alu sequences and all three SDHD deletions showed an intron 2 breakpoint. This study underlines the fact that clinically relevant deletions may encompass neighboring genes, with the potential to modify phenotype. Gene deletions of SDHD and SDHC represent a substantial proportion of all mutations, and must be considered in paraganglioma patients shown to be negative for mutations by sequencing. PMID- 19546168 TI - Integrative molecular bioinformatics study of human adrenocortical tumors: microRNA, tissue-specific target prediction, and pathway analysis. AB - MicroRNAs (miRs) are involved in the pathogenesis of several neoplasms; however, there are no data on their expression patterns and possible roles in adrenocortical tumors. Our objective was to study adrenocortical tumors by an integrative bioinformatics analysis involving miR and transcriptomics profiling, pathway analysis, and a novel, tissue-specific miR target prediction approach. Thirty-six tissue samples including normal adrenocortical tissues, benign adenomas, and adrenocortical carcinomas (ACC) were studied by simultaneous miR and mRNA profiling. A novel data-processing software was used to identify all predicted miR-mRNA interactions retrieved from PicTar, TargetScan, and miRBase. Tissue-specific target prediction was achieved by filtering out mRNAs with undetectable expression and searching for mRNA targets with inverse expression alterations as their regulatory miRs. Target sets and significant microarray data were subjected to Ingenuity Pathway Analysis. Six miRs with significantly different expression were found. miR-184 and miR-503 showed significantly higher, whereas miR-511 and miR-214 showed significantly lower expression in ACCs than in other groups. Expression of miR-210 was significantly lower in cortisol-secreting adenomas than in ACCs. By calculating the difference between dCT(miR-511) and dCT(miR-503) (delta cycle threshold), ACCs could be distinguished from benign adenomas with high sensitivity and specificity. Pathway analysis revealed the possible involvement of G2/M checkpoint damage in ACC pathogenesis. To our knowledge, this is the first report describing miR expression patterns and pathway analysis in sporadic adrenocortical tumors. miR biomarkers may be helpful for the diagnosis of adrenocortical malignancy. This tissue-specific target prediction approach may be used in other tumors too. PMID- 19546169 TI - Sequence and structural variation in a human genome uncovered by short-read, massively parallel ligation sequencing using two-base encoding. AB - We describe the genome sequencing of an anonymous individual of African origin using a novel ligation-based sequencing assay that enables a unique form of error correction that improves the raw accuracy of the aligned reads to >99.9%, allowing us to accurately call SNPs with as few as two reads per allele. We collected several billion mate-paired reads yielding approximately 18x haploid coverage of aligned sequence and close to 300x clone coverage. Over 98% of the reference genome is covered with at least one uniquely placed read, and 99.65% is spanned by at least one uniquely placed mate-paired clone. We identify over 3.8 million SNPs, 19% of which are novel. Mate-paired data are used to physically resolve haplotype phases of nearly two-thirds of the genotypes obtained and produce phased segments of up to 215 kb. We detect 226,529 intra-read indels, 5590 indels between mate-paired reads, 91 inversions, and four gene fusions. We use a novel approach for detecting indels between mate-paired reads that are smaller than the standard deviation of the insert size of the library and discover deletions in common with those detected with our intra-read approach. Dozens of mutations previously described in OMIM and hundreds of nonsynonymous single-nucleotide and structural variants in genes previously implicated in disease are identified in this individual. There is more genetic variation in the human genome still to be uncovered, and we provide guidance for future surveys in populations and cancer biopsies. PMID- 19546170 TI - Deterministic protein inference for shotgun proteomics data provides new insights into Arabidopsis pollen development and function. AB - Pollen, the male gametophyte of flowering plants, represents an ideal biological system to study developmental processes, such as cell polarity, tip growth, and morphogenesis. Upon hydration, the metabolically quiescent pollen rapidly switches to an active state, exhibiting extremely fast growth. This rapid switch requires relevant proteins to be stored in the mature pollen, where they have to retain functionality in a desiccated environment. Using a shotgun proteomics approach, we unambiguously identified approximately 3500 proteins in Arabidopsis pollen, including 537 proteins that were not identified in genetic or transcriptomic studies. To generate this comprehensive reference data set, which extends the previously reported pollen proteome by a factor of 13, we developed a novel deterministic peptide classification scheme for protein inference. This generally applicable approach considers the gene model-protein sequence-protein accession relationships. It allowed us to classify and eliminate ambiguities inherently associated with any shotgun proteomics data set, to report a conservative list of protein identifications, and to seamlessly integrate data from previous transcriptomics studies. Manual validation of proteins unambiguously identified by a single, information-rich peptide enabled us to significantly reduce the false discovery rate, while keeping valuable identifications of shorter and lower abundant proteins. Bioinformatic analyses revealed a higher stability of pollen proteins compared to those of other tissues and implied a protein family of previously unknown function in vesicle trafficking. Interestingly, the pollen proteome is most similar to that of seeds, indicating physiological similarities between these developmentally distinct tissues. PMID- 19546171 TI - Positional conservation and amino acids shape the correct diagnosis and population frequencies of benign and damaging personal amino acid mutations. AB - As the cost of DNA sequencing drops, we are moving beyond one genome per species to one genome per individual to improve prevention, diagnosis, and treatment of disease by using personal genotypes. Computational methods are frequently applied to predict impairment of gene function by nonsynonymous mutations in individual genomes and single nucleotide polymorphisms (nSNPs) in populations. These computational tools are, however, known to fail 15%-40% of the time. We find that accurate discrimination between benign and deleterious mutations is strongly influenced by the long-term (among species) history of positions that harbor those mutations. Successful prediction of known disease-associated mutations (DAMs) is much higher for evolutionarily conserved positions and for original mutant amino acid pairs that are rarely seen among species. Prediction accuracies for nSNPs show opposite patterns, forecasting impediments to building diagnostic tools aiming to simultaneously reduce both false-positive and false-negative errors. The relative allele frequencies of mutations diagnosed as benign and damaging are predicted by positional evolutionary rates. These allele frequencies are modulated by the relative preponderance of the mutant allele in the set of amino acids found at homologous sites in other species (evolutionarily permissible alleles [EPAs]). The nSNPs found in EPAs are biochemically less severe than those missing from EPAs across all allele frequency categories. Therefore, it is important to consider position evolutionary rates and EPAs when interpreting the consequences and population frequencies of human mutations. The impending sequencing of thousands of human and many more vertebrate genomes will lead to more accurate classifiers needed in real-world applications. PMID- 19546172 TI - Epigenetics of human T cells during the G0-->G1 transition. AB - We investigated functional epigenetic changes that occur in primary human T lymphocytes during entry into the cell cycle and mapped these at the single nucleosome level by ChIP-chip on tiling arrays for chromosomes 1 and 6. We show that nucleosome loss and flanking active histone marks define active transcriptional start sites (TSSs). Moreover, these signatures are already set at many inducible genes in quiescent cells prior to cell stimulation. In contrast, there is a dearth of the inactive histone mark H3K9me3 at the TSS, and under representation of H3K9me2 and H3K9me3 defines the body of active genes. At the DNA level, cytosine methylation (meC) is enriched for nucleosomes that remain at the TSS, whereas in general there is a dearth of meC at TSSs. Furthermore, a drop in meC also marks 3' transcription termination, and a peak of meC occurs at stop codons. This mimics the 3' nucleosomal distribution in yeast, which we show does not occur in human T cells. PMID- 19546174 TI - Neglect of the coronary circulation: some critical remarks on problems in the translation of cardioprotection. PMID- 19546175 TI - The effect of Premium View post-processing software on digital mammographic reporting. AB - The aim of this study was to identify the effect of the installation of Premium View post-processing software on our mammographic reporting performance, in particular the effects on our recall rate, biopsy rate and cancer detection rate. The case notes and imaging of all patients discussed at the weekly indeterminate imaging multidisciplinary team meeting were reviewed retrospectively before, immediately after and at a delayed interval following the installation of Premium View post-processing software. Factors recorded included the mammographic abnormality, further investigations and final histology. The indeterminate mammogram rate increased significantly from a baseline of 5.7% (before Premium View) to 8.7% in the time period immediately after the installation of Premium View (p=0.002). The stereotactic biopsy rate also increased from 0.8% to 2.4% (p=0.001), with a significant increase in the overall cancer detection rate from 3.4% to 4.4% (p=0.02). In the follow-up period several months after the installation of Premium View, the indeterminate mammogram rate returned to a level similar to that before Premium View (6%; p=0.7). The stereotactic biopsy rate remained significantly higher at 1.6% (p=0.07), as did the overall cancer detection rate of 5.0% (p=0.003). In conclusion, the use of Premium View may lead to higher cancer detection rates, at the expense of an initial increase in recall rate. Although prospective studies are suggested, this result is of interest in light of the proposed installation of digital mammography across the NHS Breast Screening Programme. PMID- 19546173 TI - Atrial natriuretic peptide suppresses endothelin gene expression and proliferation in cardiac fibroblasts through a GATA4-dependent mechanism. AB - AIMS: Atrial natriuretic peptide (ANP) is a hormone that has both antihypertrophic and antifibrotic properties in the heart. We hypothesized that myocyte-derived ANP inhibits endothelin (ET) gene expression in fibroblasts. METHODS AND RESULTS: We have investigated the mechanism(s) involved in the antiproliferative effect of ANP on cardiac fibroblasts in a cell culture model. We found that cardiac myocytes inhibited DNA synthesis in co-cultured cardiac fibroblasts as did treatment with the ET-1 antagonist BQ610. The effect of co culture was reversed by antibody directed against ANP or the ANP receptor antagonist HS-142-1. ANP inhibited the expression of the ET-1 gene and ET-1 gene promoter activity in cultured fibroblasts. The site of the inhibition was localized to a GATA-binding site positioned between -132 and -135 upstream from the transcription start site. GATA4 expression was demonstrated in cardiac fibroblasts, GATA4 bound the ET-1 promoter both in vitro and in vivo, and siRNA mediated knockdown of GATA4 inhibited ET-1 expression. ET-1 treatment resulted in increased levels of phospho-serine(105) GATA4 in cardiac fibroblasts and this induction was partially suppressed by co-treatment with ANP. CONCLUSION: Collectively, these findings suggest that locally produced ET-1 serves as an autocrine stimulator of fibroblast proliferation, that ANP produced in neighbouring myocytes serves as a paracrine inhibitor of this proliferation, and that the latter effect operates through a reduction in GATA4 phosphorylation and coincident reduction in GATA4-dependent transcriptional activity. PMID- 19546177 TI - Enhancement pattern of the normal facial nerve at 3.0 T temporal MRI. AB - The purpose of this study was to evaluate the enhancement pattern of the normal facial nerve at 3.0 T temporal MRI. We reviewed the medical records of 20 patients and evaluated 40 clinically normal facial nerves demonstrated by 3.0 T temporal MRI. The grade of enhancement of the facial nerve was visually scaled from 0 to 3. The patients comprised 11 men and 9 women, and the mean age was 39.7 years. The reasons for the MRI were sudden hearing loss (11 patients), Meniare's disease (6) and tinnitus (7). Temporal MR scans were obtained by fluid-attenuated inversion-recovery (FLAIR) and diffusion-weighted imaging of the brain; three dimensional (3D) fast imaging employing steady-state acquisition (FIESTA) images of the temporal bone with a 0.77 mm thickness, and pre-contrast and contrast enhanced 3D spoiled gradient record acquisition in the steady state (SPGR) of the temporal bone with a 1 mm thickness, were obtained with 3.0 T MR scanning. 40 nerves (100%) were visibly enhanced along at least one segment of the facial nerve. The enhanced segments included the geniculate ganglion (77.5%), tympanic segment (37.5%) and mastoid segment (100%). Even the facial nerve in the internal auditory canal (15%) and labyrinthine segments (5%) showed mild enhancement. The use of high-resolution, high signal-to-noise ratio (with 3 T MRI), thin-section contrast-enhanced 3D SPGR sequences showed enhancement of the normal facial nerve along the whole course of the nerve; however, only mild enhancement was observed in areas associated with acute neuritis, namely the canalicular and labyrinthine segment. PMID- 19546176 TI - Similar inhibition of platelet adhesion, P-selectin expression and plasma coagulation by ioversol, iodixanol and ioxaglate. AB - Contrast media (CM) are reported to possess both prothrombotic and anticoagulant properties. The mechanisms are not clearly understood, and early reports are contradictory. To study the effects of CM on haemostasis, we analysed the ex vivo effects of ioversol and iodixanol on platelet adhesion and P-selectin expression, and the in vitro effects of ioversol, iodixanol and ioxaglate on platelet adhesion, P-selectin expression and plasma coagulation. A novel enzymatic assay was used to measure platelet adhesion to protein surfaces, and an enzyme-linked immunosorbent assay was used to measure platelet P-selectin surface expression. Prothrombin time (PT) and activated partial thromboplastin time (APTT) were used to measure plasma coagulation. The ex vivo study consisted of blood from 27 outpatients administered ioversol and 9 patients administered iodixanol intravenously. Samples were collected before and 5 min after CM administration. Healthy donors were used for the in vitro studies on the effects of CM. The ex vivo study showed significantly (p<0.05) decreased platelet adhesion and P selectin expression after administration of ioversol and iodixanol. Adhesion was more affected than P-selectin expression. The in vitro study showed that ioversol, iodixanol and ioxaglate significantly (p<0.05) and dose-dependently (beginning at 3 mg ml(-1)) decreased platelet adhesion and P-selectin expression. APTT and PT were significantly (p<0.01) prolonged at concentrations of 10 mg ml( 1) and 30 mg ml(-1), respectively. In conclusion, ioversol, iodixanol and ioxaglate inhibit platelet adhesion and P-selectin expression, as well as plasma coagulation. Platelets are more sensitive in relation to the inhibiting effect on plasma coagulation. PMID- 19546178 TI - Does the concomitant intra-arterial injection of asialoerythropoietin and edaravone mitigate ischaemic mucosal damage after acute superior mesenteric artery thromboembolism in a rabbit autologous fibrin clot model? AB - To increase the survival rate of patients with acute superior mesenteric artery thromboembolism (ASMAT) treated by catheter thrombolysis, we examined the effects of delivering edaravone and asialoerythropoietin, agents with tissue-protective activities, using a rabbit autologous fibrin clot ASMAT model. Japanese white rabbits (n=32) were randomly separated into four equal groups. 45 min after introducing autologous fibrin clot, Group U received urokinase and heparin; Group E received urokinase and heparin plus edaravone; Group A received urokinase and heparin plus asialoerythropoietin; and Group EA received urokinase, heparin and edaravone plus asialoerythropoietin via a catheter. The intestines were removed 6 h later and intestinal mucosal damage was scored using the Park's injury score. Survival time was assessed. Average mucosal injury was 5.78+/-1.52 (Group U), 2.88+/-0.72 (Group E), 1.90+/-1.23 (Group A) and 1.18+/-1.25 (Group EA). The degree of mucosal injury was significantly lower in Group EA than in Groups U and E (p<0.05). Conversely, there was no significant difference between Group A and Group EA, or between Group A and Group E. The survival times were 31.50+/-13.30 h (Group U), 51.00+/-24.74 h (Group E), 48.00+/-16.97 h (Group A) and 82+/-51.07 h (Group EA); the difference among the four groups was not significant. In conclusion, the concomitant administration of asialoerythropoietin and edaravone reduced mucosal membrane injury significantly compared with edaravone alone. However, to improve the survival of ASMAT rabbit models, the delivery of an appropriate dose of asialoerythropoietin is required, together with the development of methods to assess peripheral recanalisation. PMID- 19546179 TI - Incidental breast lesions detected on CT: what is their significance? AB - An increasing number of breast lesions are being detected incidentally on CT. The aim of this study was to investigate the rate of referrals to the breast unit for assessment of lesions identified on CT and the resulting yield of previously undiagnosed breast malignancies from this pathway. A retrospective review was undertaken of CT examinations conducted over a period of 14 years. All patients (with no previous history of breast cancer) whose report contained the keyword "breast" and who were referred to a specialist breast unit for assessment were reviewed. CT lesion morphology and enhancement pattern were identified and compared with the final diagnostic outcome. 70 patients were identified by retrospective analysis, yielding 78 incidental breast lesions, of which 22 (28.2%) were malignant (category B5). This gave a positive predictive value (PPV) for malignancy of 28.2%. The best morphological predictor of malignancy was spiculation (PPV, 76%) and irregularity (PPV, 58%), whereas calcification patterns (PPV, 36%) were diagnostically unhelpful. Malignant lesions were likely to be larger (mean, 28.5 mm) than benign lesions (mean, 20.2 mm; p<0.05). In conclusion, 30% of incidental breast lesions in this large series of patients proved to be unsuspected breast cancers, particularly irregular spiculated masses. Referral for formal triple assessment of CT-diagnosed breast lesions is worthwhile, and careful examination of the breast should be a routine part of CT examinations. PMID- 19546180 TI - Assessment of left ventricular ejection fraction and regional wall motion with 64 slice multidetector CT: a comparison with two-dimensional transthoracic echocardiography. AB - The aim of this study was to compare the measurement of left ventricular ejection fraction (LVEF) and regional wall motion using 64-slice multidetector CT (MDCT) with that using two-dimensional transthoracic echocardiography (2D-TTE) in a heterogeneous patient population. In 126 patients with angina pectoris, acute myocardial infarction, chronic myocardial infarction, atypical chest pain without coronary artery disease or valvular heart disease, 64-slice MDCT was performed using retrospective electrocardiography gating without dose modulation. 20 phases of the cardiac cycle were analysed to identify the end-diastolic and end-systolic phases and to assess regional LV wall motion. For these measurements, 2D-TTE served as the reference standard. MDCT and 2D-TTE were performed within 10 days of each other. An excellent correlation between MDCT and 2D-TTE was shown for the evaluation of LVEF (59.2+/-11% vs 57.9+/-10%, respectively; r = 0.87). LVEF was slightly overestimated by MDCT, when compared with 2D-TTE, by an average of 1.4+/ 5.6%. Good agreement was obtained between the use of the two techniques, with 94% of the segments scored identically on both modalities (kappa = 0.70). MDCT had a sensitivity of 97% and a specificity of 82% when compared with 2D-TTE as the reference standard. In conclusion, the use of 64-slice MDCT can provide comparable results to those using 2D-TTE for LVEF and regional wall motion assessment in a heterogeneous population. PMID- 19546181 TI - Correlation of intra-articular osseous measurements with posterior cruciate ligament length on MRI scans. AB - Six patients with a clinical diagnosis of chronic posterior cruciate ligament (PCL) rupture, based on a positive posterior drawer test, had a normal appearance of the PCL on an MRI scan. It is postulated that the PCL had been ruptured but healed in a lengthened state. 12 volunteers with no history of knee trauma underwent an MRI scan of the knee. In this control group (n = 12), there was a close correlation between the lateral femoral condylar width in the sagittal plane and the PCL length, with a ratio of 2:1 (95% confidence interval (CI) = 1.817-2.095). In the clinically abnormal group (n = 6), the ratio was 1.49:1 (95% CI = 1.206-1.782) (p< 0.0005). In conclusion, the ratio of the lateral femoral condylar width in the sagittal plane to the PCL length is a useful index for diagnosing PCL attenuation and lengthening in the presence of a normal morphological MR appearance. PMID- 19546182 TI - How safe is the antiarrhythmic drug therapy in atrial fibrillation? PMID- 19546183 TI - The risk of antiarrhythmic drugs in atrial fibrillation: 20 years of controversies. PMID- 19546184 TI - Antithrombotic therapy use for patients with atrial fibrillation undergoing percutaneous coronary intervention: new data in an area of limited evidence. PMID- 19546185 TI - Enhanced efficacy with MgSO4 due to an additive, alternative, or dual mode of action? PMID- 19546186 TI - Will we be able to predict in which atrial fibrillation patients a rhythm control strategy will be successful? PMID- 19546187 TI - Automatic left ventricular pacing management: not only a technical step up. PMID- 19546188 TI - High take-off left inferior pulmonary vein as an obstacle in creating a conduction block at the lateral mitral isthmus. AB - AIMS: Creation of complete linear lesions in the lateral mitral isthmus (LMI) by catheter ablation for treating atrial fibrillation remains technically challenging. We aimed to clarify whether a high take-off left inferior pulmonary vein (LIPV) can hamper the creation of a complete block at the LMI. METHODS AND RESULTS: We included 81 consecutive patients who underwent linear ablation at the LMI and cardiac computed tomography (CT) before ablation. We defined a high take off LIPV when the level of the lower edge of the LIPV ostium was higher than that of the top of mitral annulus on CT. The clinical backgrounds, parameters, and long-term follow-up were then compared between the success (successful creation of a complete LMI block) and failure groups. A complete LMI block was obtained in 60/81 (76%) patients. In the failure group, a high take-off LIPV was noted more commonly and the LMI tended to be longer than the success group. Multivariate analysis revealed that a high take-off LIPV was an independent predictor of failure to achieve a complete LMI block. The sinus rhythm maintenance rate was not different between the success and failure groups. CONCLUSION: A high take-off LIPV hampered the creation of complete linear lesions in the LMI. PMID- 19546189 TI - The influence of fat thickness on the human epicardial bipolar electrogram characteristics: measurements on patients undergoing open-heart surgery. AB - AIMS: Reference values exist for endocardial but not for epicardial (EPI) substrate mapping in cases of cardiomyopathy-associated ventricular tachycardia. We sought to establish such values for EPI electrogram voltage, including areas with overlying fat. METHODS AND RESULTS: Ten patients (six males) undergoing cardiac surgery were studied. After opening the pericardium, the distal bipole of an electrophysiology catheter was placed tangential to the EPI surface to obtain an electrogram recording. The bipole was tangentially rotated 90 degrees and the higher of the two amplitudes (mV) was taken as the local amplitude. Recordings were taken from normal left and right ventricular myocardium (n = 26 data points each), over thick (> or = 0.5 cm) fat at both ventricular bases (n = 16) and thin (<0.5 cm) fat at the mid-ventricular level (n = 32). A total of 100 recordings (mean 10/patient) were analysed. Four patients underwent valvular surgery, three bypass surgery, and three combined procedures. Mean age was 61.7 +/- 10.4 years and mean left ventricular ejection fraction was 46 +/- 12%. Electrogram amplitude was inversely related to EPI fat thickness. Over thick fat, 31% of recordings were <0.5 mV. CONCLUSION: Human EPI electrogram amplitude varies by ventricular chamber and significantly by EPI fat thickness. A cut-off of 0.5 mV to define 'scar' will include normal areas with thick overlying fat. EPI substrate maps should include data on EPI fat thickness for higher specificity. PMID- 19546190 TI - Outer membrane protein expression profile in Helicobacter pylori clinical isolates. AB - The gram-negative gastric pathogen Helicobacter pylori is equipped with an extraordinarily large set of outer membrane proteins (OMPs), whose role in the infection process is not well understood. The Hop (Helicobacter outer membrane porins) and Hor (Hop-related proteins) groups constitute a large paralogous family consisting of 33 members. The OMPs AlpA, AlpB, BabA, SabA, and HopZ have been identified as adhesins or adherence-associated proteins. To better understand the relevance of these and other OMPs during infection, we analyzed the expression of eight different omp genes (alpA, alpB, babA, babB, babC, sabA, hopM, and oipA) in a set of 200 patient isolates, mostly from symptomatic children or young adults. Virtually all clinical isolates produced the AlpA and AlpB proteins, supporting their essential function. All other OMPs were produced at extremely variable rates, ranging from 35% to 73%, indicating a function in close adaptation to the individual host or gastric niche. In 11% of the isolates, BabA was produced, and SabA was produced in 5% of the isolates, but the strains failed to bind their cognate substrates. Interleukin-8 (IL-8) expression in gastric cells was strictly dependent on the presence of the cag pathogenicity island, whereas the presence of OipA clearly enhanced IL-8 production. The presence of the translocated effector protein CagA correlated well with BabA and OipA production. In conclusion, we found unexpectedly diverse omp expression profiles in individual H. pylori strains and hypothesize that this reflects the selective pressure for adhesion, which may differ across different hosts as well as within an individual over time. PMID- 19546191 TI - Functional and immunological characterization of a Duffy binding-like alpha domain from Plasmodium falciparum erythrocyte membrane protein 1 that mediates rosetting. AB - The Duffy binding-like (DBL) domains are common adhesion modules present in Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) variants, which are responsible for immune evasion and cytoadherence. Knowledge about how immune responses are acquired against polymorphic DBL domains of PfEMP1 can aid in the development of vaccines for malaria. A recombinant DBLalpha domain, encoded by R29 var1, which binds complement receptor 1 to mediate rosetting by the P. falciparum laboratory strain R29, was expressed in Escherichia coli, renatured by oxidative refolding to its native form, and purified to homogeneity. Antibody levels in 704 plasmas obtained from residents of areas of different levels of malaria endemicity in Orissa (India) and Manhica (Mozambique) were assessed by enzyme-linked immunosorbent assay. The refolded DBLalpha domain was pure, homogeneous, and functional in that it bound human erythrocytes with specificity and was capable of inhibiting rosetting. The proportion of individuals who had measurable anti-DBLalpha immunoglobulin G responses was low in areas of low malaria endemicity in Orissa (6.7%) but high in areas of high endemicity in Orissa (87.5%) and Manhica (74.5%). Seroprevalence and antibody levels against the recombinant protein increased with the age of inhabitants from areas with high transmission rates (P < 0.001). Half of the children in these areas had seroconverted by the age of 5 years. These findings suggest that in spite of the extreme polymorphism of PfEMP1 DBLalpha domains, the acquisition of specific antibodies is rapid and age related and reflects the reduced risk of malaria in areas with high transmission rates. Further studies are required to elucidate the role of these antibodies in protection from malaria. PMID- 19546192 TI - Neisseria gonorrhoeae infection protects human endocervical epithelial cells from apoptosis via expression of host antiapoptotic proteins. AB - Several microbial pathogens can modulate the host apoptotic response to infection, which may contribute to immune evasion. Various studies have reported that infection with the sexually transmitted disease pathogen Neisseria gonorrhoeae can either inhibit or induce apoptosis. N. gonorrhoeae infection initiates at the mucosal epithelium, and in women, cells from the ectocervix and endocervix are among the first host cells encountered by this pathogen. In this study, we defined the antiapoptotic effect of N. gonorrhoeae infection in human endocervical epithelial cells (End/E6E7 cells). We first established that N. gonorrhoeae strain FA1090B failed to induce cell death in End/E6E7 cells. Subsequently, we demonstrated that stimulation with N. gonorrhoeae protected these cells from staurosporine (STS)-induced apoptosis. Importantly, only End/E6E7 cells incubated with live bacteria and in direct association with N. gonorrhoeae were protected from STS-induced apoptosis, while heat-killed and antibiotic-killed bacteria failed to induce protection. Stimulation of End/E6E7 cells with live N. gonorrhoeae induced NF-kappaB activation and resulted in increased gene expression of the NF-kappaB-regulated antiapoptotic genes bfl-1, cIAP-2, and c-FLIP. Furthermore, cIAP-2 protein levels also increased in End/E6E7 cells incubated with gonococci. Collectively, our results indicate that the antiapoptotic effect of N. gonorrhoeae in human endocervical epithelial cells results from live infection via expression of host antiapoptotic proteins. Securing an intracellular niche through the inhibition of apoptosis may be an important mechanism utilized by N. gonorrhoeae for microbial survival and immune evasion in cervical epithelial cells. PMID- 19546193 TI - Decreased expression of colonic Slc26a3 and carbonic anhydrase iv as a cause of fatal infectious diarrhea in mice. AB - Citrobacter rodentium causes epithelial hyperplasia and colitis and is used as a model for enteropathogenic and enterohemorrhagic Escherichia coli infections. Little or no mortality develops in most inbred strains of mice, but C3H and FVB/N mice exhibit fatal outcomes of infection. Here we test the hypothesis that decreased intestinal transport activity during C. rodentium infection results in fatality in C3H/HeOu and FVB/N mice. Susceptible strains were compared to resistant C57BL/6 mice and to inbred strains SWR and SJL of Swiss origin, which have not been previously characterized for outcomes of C. rodentium infection. Mortality in susceptible strains C3H/HeOu and FVB/N was associated with significant fluid loss in feces, a remarkable downregulation of Slc26a3 and carbonic anhydrase IV (CAIV) message and protein expression, retention of chloride in stool, and hypochloremia, suggesting defects in intestinal chloride absorption. SWR, SJL, and C57BL/6 mice were resistant and survived the infection. Fluid therapy fully prevented mortality in C3H/HeOu and FVB/N mice without affecting clinical disease. Common pathogenic mechanisms, such as decreased levels of expression of Slc26a3 and CAIV, affect intestinal ion transport in C. rodentium-infected FVB and C3H mice, resulting in profound electrolyte loss, dehydration, and mortality. Intestinal chloride absorption pathways are likely a potential target for the treatment of infectious diarrhea. PMID- 19546194 TI - Amino acid changes in elongation factor Tu of Mycoplasma pneumoniae and Mycoplasma genitalium influence fibronectin binding. AB - Mycoplasma pneumoniae and Mycoplasma genitalium are closely related organisms that cause distinct clinical manifestations and possess different tissue predilections despite their high degree of genome homology. We reported earlier that surface-localized M. pneumoniae elongation factor Tu (EF-Tu(Mp)) mediates binding to the extracellular matrix component fibronectin (Fn) through the carboxyl region of EF-Tu. In this study, we demonstrate that surface-associated M. genitalium EF-Tu (EF-Tu(Mg)), in spite of sharing 96% identity with EF-Tu(Mp), does not bind Fn. We utilized this finding to identify the essential amino acids of EF-Tu(Mp) that mediate Fn interactions by generating modified recombinant EF Tu proteins with amino acid changes corresponding to those of EF-Tu(Mg). Amino acid changes in serine 343, proline 345, and threonine 357 were sufficient to significantly reduce the Fn binding of EF-Tu(Mp). Synthetic peptides corresponding to this region of EF-Tu(Mp) (EF-Tu(Mp) 340-358) blocked both recombinant EF-Tu(Mp) and radiolabeled M. pneumoniae cell binding to Fn. In contrast, EF-Tu(Mg) 340-358 peptides exhibited minimal blocking activity, reinforcing the specificity of EF-Tu-Fn interactions as mediators of microbial colonization and tissue tropism. PMID- 19546195 TI - Real-time observation of listeria monocytogenes-phagocyte interactions in living zebrafish larvae. AB - The zebrafish, Danio rerio, has become a popular vertebrate model for the study of infections, mainly because of its excellent optical accessibility at the embryonic and larval stages, when the innate immune system is already effective. We have thus tested the susceptibility of zebrafish larvae to the human pathogen Listeria monocytogenes, a gram-positive, facultative, intracellular bacterium that is known to survive and multiply in professional phagocytes and that causes fatal meningitis and abortions. Intravenous injection of early zebrafish larvae resulted in a progressive and ultimately fatal infection. Blood-borne L. monocytogenes bacteria were quickly trapped and engulfed by macrophages, an event that, for the first time, could be captured in vivo and in real time. Granulocytes also participated in the innate immune response. As in mammals, bacteria could escape the macrophage phagosome in a listeriolysin-dependent manner and accessed the cytosol; this event was critical for bacterial virulence, as listeriolysin-deficient bacteria were completely avirulent. Actin comet tails and protrusions were observed, suggesting cell-to-cell spread; these phenomena also played a role in virulence in zebrafish larvae, as actA-deficient bacteria were attenuated. These results demonstrate the relevance of the genetically tractable and optically accessible zebrafish model for the study of L. monocytogenes pathogenesis and particularly for the dissection of its interactions with phagocytes in vivo, a key factor of L. monocytogenes virulence. PMID- 19546196 TI - Oligoclonal CD4+ T cells promote host memory immune responses to Zwitterionic polysaccharide of Streptococcus pneumoniae. AB - Zwitterionic polysaccharides of the normal flora bacteria represent a novel class of antigens in that they correct systemic CD4(+) T-cell deficiencies and direct lymphoid organogenesis during colonization of the host. Presentation of these polysaccharides to CD4(+) T cells depends on major histocompatibility complex class II- and DM-dependent retrograde transport from lysosomes to the cell surface. Yet the phenotype and clonality of the immune response to the polysaccharide in the mature host immune system have not been studied. Using the zwitterionic capsular polysaccharide Sp1 of Streptococcus pneumoniae, a transient member of the bacterial flora, in an experimental mouse model of cellular immunity, we demonstrated the accumulation of TH1- and TH17-polarized CD4(+) CD44(high) CD62(low) CD25(-) memory T cells. Subcutaneous immunization with Sp1 resulted in an increase of serum immunoglobulin G (IgG), predominantly of the IgG1 subclass, and suggested the presence of a humoral memory response to the polysaccharide. CD4(+) T cells stimulated with polysaccharide in vitro and in vivo showed a nonrestricted pattern for the T-cell receptor (TCR) beta-chain variable region, as demonstrated by semiquantitative reverse transcription-PCR and flow cytometry. Clonotype mapping of in vivo and in vitro polysaccharide activated CD4(+) T cells revealed clonotypic TCR transcripts. Taken together, the data show the induction of clonal expansion of CD4(+) T cells by polysaccharides of commensal bacteria. Cellular and humoral memory host responses imply the ability of these polysaccharides to mediate the expansion of T cells via recognition within the CDR3 region of the TCR. PMID- 19546198 TI - Geographic information systems (GIS) for Health Promotion and Public Health: a review. AB - The purpose of this literature review is to identify how geographic information system (GIS) applications have been used in health-related research and to critically examine the issues, strengths, and challenges inherent to those approaches from the lenses of health promotion and public health. Through the review process, conducted in 2007, it is evident that health promotion and public health applications of GIS can be generally categorized into four predominant themes: disease surveillance (n = 227), risk analysis (n = 189), health access and planning (n = 138), and community health profiling (n = 115). This review explores how GIS approaches have been used to inform decision making and discusses the extent to which GIS can be applied to address health promotion and public health questions. The contribution of this literature review will be to generate a broader understanding of how GIS-related methodological techniques and tools developed in other disciplines can be meaningfully applied to applications in public health policy, promotion, and practice. PMID- 19546197 TI - Injection of Staphylococcus aureus EDIN by the Bacillus anthracis protective antigen machinery induces vascular permeability. AB - Systemic injection of Bacillus anthracis lethal toxin (LT) produces vascular leakage and animal death. Recent studies suggest that LT triggers direct endothelial cell cytotoxicity that is responsible for the vascular leakage. LT is composed of heptamers of protective antigen (PA), which drives the endocytosis and translocation into host cells of the lethal factor (LF), a mitogen-activated protein kinase kinase protease. Here we investigated the consequences of injection of an endothelium-permeabilizing factor using LT as a "molecular syringe." To this end, we generated the chimeric factor LE, corresponding to the PA-binding domain of LF (LF(1-254)) fused to EDIN exoenzyme. EDIN ADP ribosylates RhoA, leading to actin cable disruption and formation of transcellular tunnels in endothelial cells. We report that systemic injection of LET (LE plus PA) triggers a PA-dependent increase in the pulmonary endothelium permeability. We also report that native LT induces a progressive loss of endothelium barrier function. We established that there is a direct correlation between the extent of endothelium permeability induced by LT and the cytotoxic activity of LT. This suggests new ways to design therapeutic drugs against anthrax directed toward vascular permeability. PMID- 19546199 TI - Advances in pharmacology and therapeutics. PMID- 19546200 TI - Advances in patient comfort: awake, delirious, or restrained. PMID- 19546201 TI - Neurokinin-1 antagonists: a step change in prevention of postoperative nausea and vomiting? PMID- 19546202 TI - Current concepts in neuromuscular transmission. AB - The neuromuscular junction (NMJ) is structured and powered to transduce electrical activity from the distal nerve terminal of a motor neurone via the neuromuscular cleft to the post-junctional muscle membrane to ultimately generate muscle contraction. Our understanding of this complex function has expanded over many years, and the NMJ has served as a prototype for how different synapses operate in the peripheral and central nervous systems. The NMJ has a presynaptic part which is synonymous with the distal nerve ending, being responsible for neurotransmitter synthesis, packaging into vesicles, and subsequent vesicle transportation to active release sites where vesicle docking, fusion, and release of acetylcholine and other co-released transmitters finally take place. The synaptic cleft, filled with large molecular complexes that guarantee ultrastructural NMJ arrangement and signal transduction, allows for rapid diffusion and degradation of the neurotransmitter. The postsynaptic part consists of a folded muscle membrane into which nicotinic acetylcholine receptors (nAChRs) directly opposite the presynaptic active release sites are mounted and fixed by a cytoskeleton. This specialized postsynaptic region is closely associated with the perijunctional zone where a high density of sodium channels promote and amplify the signal in order to guarantee the propagation of the electrical activity to generate muscle contraction. The transduction process is maintained at load (i.e. high stimulus frequency) by a presynaptic mechanism allowing for sustained transmitter release over time at high demand. This positive feedback mechanism relies on neuronal nAChRs present on the distal nerve terminal, whereas the continuation of the transduction process at the postsynaptic part relies on the classical muscle type nAChR. In this review, we will focus on recent findings of potential clinical importance that will advance our understanding of the effects of neuromuscular blocking agents and neuromuscular monitoring and also our management of disorders of the neuromuscular system within anaesthesia and intensive care. PMID- 19546203 TI - Major complications of central neuraxial block: the Third National Audit Project: some comments and questions. PMID- 19546204 TI - Bupivacaine chondrotoxicity. PMID- 19546205 TI - Video laryngoscopy and external laryngeal manipulation. PMID- 19546206 TI - Bispectral index sensor as a possible cause of postoperative visual loss after frontal craniotomy. PMID- 19546207 TI - Response entropy-state entropy difference and nociception: a matter of context. PMID- 19546208 TI - Diagnosis of vertebral canal haematoma by myelography and spiral computer tomography in a patient with an implantable cardioverter-defibrillator contraindicating magnetic resonance imaging. PMID- 19546209 TI - Incidental recognition of an aspirated tablet in an oesophagectomized patient. PMID- 19546210 TI - Familial Mediterranean fever abdominal pain during spinal anaesthesia. PMID- 19546211 TI - Helicobacter pylori CagA causes mitotic impairment and induces chromosomal instability. AB - Infection with cagA-positive Helicobacter pylori is the strongest risk factor for the development of gastric carcinoma. The cagA gene product CagA, which is delivered into gastric epithelial cells, specifically binds to and aberrantly activates SHP-2 oncoprotein. CagA also interacts with and inhibits partitioning defective 1 (PAR1)/MARK kinase, which phosphorylates microtubule-associated proteins to destabilize microtubules and thereby causes epithelial polarity defects. In light of the notion that microtubules are not only required for polarity regulation but also essential for the formation of mitotic spindles, we hypothesized that CagA-mediated PAR1 inhibition also influences mitosis. Here, we investigated the effect of CagA on the progression of mitosis. In the presence of CagA, cells displayed a delay in the transition from prophase to metaphase. Furthermore, a fraction of the CagA-expressing cells showed spindle misorientation at the onset of anaphase, followed by chromosomal segregation with abnormal division axis. The effect of CagA on mitosis was abolished by elevated PAR1 expression. Conversely, inhibition of PAR1 kinase elicited mitotic delay similar to that induced by CagA. Thus, CagA-mediated inhibition of PAR1, which perturbs microtubule stability and thereby causes microtubule-based spindle dysfunction, is involved in the prophase/metaphase delay and subsequent spindle misorientation. Consequently, chronic exposure of cells to CagA induces chromosomal instability. Our findings reveal a bifunctional role of CagA as an oncoprotein: CagA elicits uncontrolled cell proliferation by aberrantly activating SHP-2 and at the same time induces chromosomal instability by perturbing the microtubule-based mitotic spindle. The dual function of CagA may cooperatively contribute to the progression of multistep gastric carcinogenesis. PMID- 19546212 TI - WaaA of the hyperthermophilic bacterium Aquifex aeolicus is a monofunctional 3 deoxy-D-manno-oct-2-ulosonic acid transferase involved in lipopolysaccharide biosynthesis. AB - The hyperthermophile Aquifex aeolicus belongs to the deepest branch in the bacterial genealogy. Although it has long been recognized that this unique Gram negative bacterium carries genes for different steps of lipopolysaccharide (LPS) formation, data on the LPS itself or detailed knowledge of the LPS pathway beyond the first committed steps of lipid A and 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) synthesis are still lacking. We now report the functional characterization of the thermostable Kdo transferase WaaA from A. aeolicus and provide evidence that the enzyme is monofunctional. Compositional analysis and mass spectrometry of purified A. aeolicus LPS, showing the incorporation of a single Kdo residue as an integral component of the LPS, implicated a monofunctional Kdo transferase in LPS biosynthesis of A. aeolicus. Further, heterologous expression of the A. aeolicus waaA gene in a newly constructed Escherichia coli DeltawaaA suppressor strain resulted in synthesis of lipid IVA precursors substituted with one Kdo sugar. When highly purified WaaA of A. aeolicus was subjected to in vitro assays using mass spectrometry for detection of the reaction products, the enzyme was found to catalyze the transfer of only a single Kdo residue from CMP-Kdo to differently modified lipid A acceptors. The Kdo transferase was capable of utilizing a broad spectrum of acceptor substrates, whereas surface plasmon resonance studies indicated a high selectivity for the donor substrate. PMID- 19546213 TI - Hypoxia-inducible factor prolyl-4-hydroxylase PHD2 protein abundance depends on integral membrane anchoring of FKBP38. AB - Prolyl-4-hydroxylase domain (PHD) proteins are 2-oxoglutarate and dioxygen dependent enzymes that mediate the rapid destruction of hypoxia-inducible factor alpha subunits. Whereas PHD1 and PHD3 proteolysis has been shown to be regulated by Siah2 ubiquitin E3 ligase-mediated polyubiquitylation and proteasomal destruction, protein regulation of the main oxygen sensor responsible for hypoxia inducible factor alpha regulation, PHD2, remained unknown. We recently reported that the FK506-binding protein (FKBP) 38 specifically interacts with PHD2 and determines PHD2 protein stability in a peptidyl-prolyl cis-trans isomerase independent manner. Using peptide array binding assays, fluorescence spectroscopy, and fluorescence resonance energy transfer analysis, we defined a minimal linear glutamate-rich PHD2 binding domain in the N-terminal part of FKBP38 and showed that this domain forms a high affinity complex with PHD2. Vice versa, PHD2 interacted with a non-linear N-terminal motif containing the MYND (myeloid, Nervy, and DEAF-1)-type Zn(2+) finger domain with FKBP38. Biochemical fractionation and immunofluorescence analysis demonstrated that PHD2 subcellular localization overlapped with FKBP38 in the endoplasmic reticulum and mitochondria. An additional fraction of PHD2 was found in the cytoplasm. In cellulo PHD2/FKBP38 association, as well as regulation of PHD2 protein abundance by FKBP38, is dependent on membrane- anchored FKBP38 localization mediated by the C-terminal transmembrane domain. Mechanistically our data indicate that PHD2 protein stability is regulated by a ubiquitin-independent proteasomal pathway involving FKBP38 as adaptor protein that mediates proteasomal interaction. We hypothesize that FKBP38-bound PHD2 is constantly degraded whereas cytosolic PHD2 is stable and able to function as an active prolyl-4-hydroxylase. PMID- 19546214 TI - A functional P2X7 splice variant with an alternative transmembrane domain 1 escapes gene inactivation in P2X7 knock-out mice. AB - The ATP-activated P2X7 receptor channel is involved in immune function and inflammatory pain and represents an important drug target. Here we describe a new P2X7 splice variant (P2X7(k)), containing an alternative intracellular N terminus and first transmembrane domain encoded by a novel exon 1 in the rodent P2rx7 gene. Whole cell patch clamp recordings of the rat isoform expressed in HEK293 cells revealed an 8-fold higher sensitivity to the agonist Bz-ATP and much slower deactivation kinetics when compared with the P2X7(a) receptor. Permeability measurements in Xenopus oocytes show a high permeability for N-methyl-D-glucamine immediately upon activation, suggesting that the P2X7(k) channel is constitutively dilated upon opening. The rates of agonist-induced dye uptake and membrane blebbing in HEK cells were also increased. PCR analyses and biochemical analysis by SDS-PAGE and BN-PAGE indicate that the P2X7(k) variant escapes gene deletion in one of the available P2X7(-/-) mice strains and is strongly expressed in the spleen. Taken together, we describe a novel P2X7 isoform with distinct functional properties that contributes to the diversity of P2X7 receptor signaling. Its presence in one of the P2X7(-/-) strains has important implications for our understanding of the role of this receptor in health and disease. PMID- 19546215 TI - Structural investigation of the interaction between LolA and LolB using NMR. AB - Lipoproteins that play critical roles in various cellular functions of Gram negative bacteria are localized in the cells inner and outer membranes. Lol proteins (LolA, LolB, LolC, LolD, and LolE) are involved in the transportation of outer membrane-directed lipoproteins from the inner to the outer membrane. LolA is a periplasmic chaperone that transports lipoproteins, and LolB is an outer membrane receptor that accepts lipoproteins. To clarify the structural basis for the lipoprotein transfer from LolA to LolB, we examined the interaction between LolA and mLolB, a soluble mutant of LolB, using solution NMR spectroscopy. We determined the interaction mode between LolA and mLolB with conformational changes of LolA. Based upon the observations, we propose that the LolA.LolB complex forms a tunnel-like structure, where the hydrophobic insides of LolA and LolB are connected, which enables lipoproteins to transfer from LolA to LolB. PMID- 19546216 TI - Loss of parkin or PINK1 function increases Drp1-dependent mitochondrial fragmentation. AB - Loss-of-function mutations in the parkin gene (PARK2) and PINK1 gene (PARK6) are associated with autosomal recessive parkinsonism. PINK1 deficiency was recently linked to mitochondrial pathology in human cells and Drosophila melanogaster, which can be rescued by parkin, suggesting that both genes play a role in maintaining mitochondrial integrity. Here we demonstrate that an acute down regulation of parkin in human SH-SY5Y cells severely affects mitochondrial morphology and function, a phenotype comparable with that induced by PINK1 deficiency. Alterations in both mitochondrial morphology and ATP production caused by either parkin or PINK1 loss of function could be rescued by the mitochondrial fusion proteins Mfn2 and OPA1 or by a dominant negative mutant of the fission protein Drp1. Both parkin and PINK1 were able to suppress mitochondrial fragmentation induced by Drp1. Moreover, in Drp1-deficient cells the parkin/PINK1 knockdown phenotype did not occur, indicating that mitochondrial alterations observed in parkin- or PINK1-deficient cells are associated with an increase in mitochondrial fission. Notably, mitochondrial fragmentation is an early phenomenon upon PINK1/parkin silencing that also occurs in primary mouse neurons and Drosophila S2 cells. We propose that the discrepant findings in adult flies can be explained by the time of phenotype analysis and suggest that in mammals different strategies may have evolved to cope with dysfunctional mitochondria. PMID- 19546217 TI - N-cadherin/p120 catenin association at cell-cell contacts occurs in cholesterol rich membrane domains and is required for RhoA activation and myogenesis. AB - p120 catenin is a major regulator of cadherin stability at cell-cell contacts and a modulator of Rho GTPase activities. In C2C12 myoblasts, N-cadherin is stabilized at cell contacts through its association with cholesterol-rich membrane domains or lipid rafts (LR) and acts as an adhesion-activated receptor that activates RhoA, an event required for myogenesis induction. Here, we report that association of p120 catenin with N-cadherin at cell contacts occurs specifically in LR. We demonstrate that interaction of p120 catenin with N cadherin is required for N-cadherin association with LR and for its stabilization at cell contacts. LR disruption inhibits myogenesis induction and N-cadherin dependent RhoA activation as does the perturbation of the N-cadherin-p120 catenin complex after p120 catenin knockdown. Finally, we observe an N-cadherin-dependent accumulation of RhoA at phosphatidylinositol 4,5-bisphosphate-enriched cell contacts which is lost after LR disruption. Thus, a functional N-cadherin-catenin complex occurs in cholesterol-rich membrane microdomains which allows the recruitment of RhoA and the regulation of its activity during myogenesis induction. PMID- 19546218 TI - Transient oxidative stress damages mitochondrial machinery inducing persistent beta-cell dysfunction. AB - Transient exposure of beta-cells to oxidative stress interrupts the transduction of signals normally coupling glucose metabolism to insulin secretion. We investigated putative persistence of effects induced by one transient oxidative stress (200 microm H(2)O(2), 10 min) on insulin secreting cells following recovery periods of days and weeks. Three days after oxidative stress INS-1E cells and rat islets exhibited persistent dysfunction. In particular, the secretory response to 15 mm glucose was reduced by 40% in INS-1E cells stressed 3 days before compared with naive cells. Compared with non-stressed INS-1E cells, we observed reduced oxygen consumption (-43%) and impaired glucose-induced ATP generation (-46%). These parameters correlated with increased mitochondrial reactive oxygen species formation (+60%) accompanied with down-regulation of subunits of the respiratory chain and decreased expression of genes responsible for mitochondrial biogenesis (TFAM, -24%; PGC-1alpha, -67%). Three weeks after single oxidative stress, both mitochondrial respiration and secretory responses were recovered. Moreover, such recovered INS-1E cells exhibited partial resistance to a second transient oxidative stress and up-regulation of UCP2 (+78%) compared with naive cells. In conclusion, one acute oxidative stress induces beta-cell dysfunction lasting over days, explained by persistent damages in mitochondrial components. PMID- 19546219 TI - Prion protein-detergent micelle interactions studied by NMR in solution. AB - Cellular prion proteins, PrP(C), carrying the amino acid substitutions P102L, P105L, or A117V, which confer increased susceptibility to human transmissible spongiform encephalopathies, are known to form structures that include transmembrane polypeptide segments. Herein, we investigated the interactions between dodecylphosphocholine micelles and the polypeptide fragments 90-231 of the recombinant mouse PrP variants carrying the amino acid replacements P102L, P105L, A117V, A113V/A115V/A118V, K110I/H111I, M129V, P105L/M129V, and A117V/M129V. Wild-type mPrP-(90-231) and mPrP[M129V]-(91-231) showed only weak interactions with dodecylphosphocholine micelles in aqueous solution at pH 7.0, whereas discrete interaction sites within the polypeptide segment 102-127 were identified for all other aforementioned mPrP variants by NMR chemical shift mapping. These model studies thus provide evidence that amino acid substitutions within the polypeptide segment 102-127 affect the interactions of PrP(C) with membranous structures, which might in turn modulate the physiological function of the protein in health and disease. PMID- 19546220 TI - ST14 (suppression of tumorigenicity 14) gene is a target for miR-27b, and the inhibitory effect of ST14 on cell growth is independent of miR-27b regulation. AB - MicroRNAs are noncoding, endogenous small RNAs that regulate target genes by cleavage of the targeted mRNA or translational repression. We investigated the microRNAome using 2-color microarrays in a highly invasive human breast cancer cell line, MDA-MB-231 (subline 4175) and a noninvasive breast epithelial cell line, MCF10A. We found 13 microRNAs that were up-regulated, and nine were down regulated significantly in 4175 cells (p < 0.05, -fold change >2) compared with MCF10A cells. Interestingly, miR-27b and its putative target gene, ST14 (suppressor of tumorigenicity 14), had inverse expression pattern in breast cancer cells. The 3'-untranslated region of ST14 contains a regulatory element for miR-27b, and our luciferase experiments indicate that antisense miR-27b enhances ST14 expression in cancer cells. Furthermore, antagomir of miR-27b suppressed cell invasion in 4175 cells, whereas pre-miR-27b stimulated invasion in moderately invasive ZR75 breast cancer cells. In addition, ST14 reduces cell proliferation as well as cell migration and invasion. Analysis of human breast tumors revealed that miR-27b expression increases during cancer progression, paralleling a decrease in ST14 expression. Furthermore, our data indicate that ST14 inhibits cells from entering into S phase by up-regulating p27, which results in down-regulation of cyclin E-CDK2 complexes, suggesting ST14 reduces cell growth through its effects on cell cycle-related proteins. Introduction of miR-27b into ST14-expressing cells did not suppress the effect on cell growth. These findings suggest that ST14 plays an important role in several biological processes, and some effects are not completely dependent on miR-27b regulation. PMID- 19546221 TI - P2X7-mediated increased intracellular calcium causes functional derangement in Schwann cells from rats with CMT1A neuropathy. AB - Charcot-Marie-Tooth (CMT) is the most frequent inherited neuromuscular disorder, affecting 1 person in 2500. CMT1A, the most common form of CMT, is usually caused by a duplication of chromosome 17p11.2, containing the PMP22 (peripheral myelin protein-22) gene; overexpression of PMP22 in Schwann cells (SC) is believed to cause demyelination, although the underlying pathogenetic mechanisms remain unclear. Here we report an abnormally high basal concentration of intracellular calcium ([Ca(2+)](i)) in SC from CMT1A rats. By the use of specific pharmacological inhibitors and through down-regulation of expression by small interfering RNA, we demonstrate that the high [Ca(2+)](i) is caused by a PMP22 related overexpression of the P2X7 purinoceptor/channel leading to influx of extracellular Ca(2+) into CMT1A SC. Correction of the altered [Ca(2+)](i) in CMT1A SC by small interfering RNA or with pharmacological inhibitors of P2X7 restores functional parameters of SC (migration and release of ciliary neurotrophic factor), which are typically defective in CMT1A SC. More significantly, stable down-regulation of the expression of P2X7 restores myelination in co-cultures of CMT1A SC with dorsal root ganglion sensory neurons. These results establish a pathogenetic link between high [Ca(2+)](i) and impaired SC function in CMT1A and identify overexpression of P2X7 as the molecular mechanism underlying both abnormalities. The development of P2X7 inhibitors is expected to provide a new therapeutic strategy for treatment of CMT1A neuropathy. PMID- 19546223 TI - Ca2+-dependent conformational changes in a C-terminal cytosolic domain of polycystin-2. AB - The PKD1 and PKD2 genes are the genes that are mutated in patients suffering from autosomal dominant polycystic kidney disease. The human PKD2 gene codes for a 968 amino acid long membrane protein called polycystin-2 that represents a cation channel whose activity can be regulated by Ca(2+) ions. By CD, fluorescence, and NMR spectroscopy, we have studied a 117-amino acid-long fragment of the cytoplasmic domain of polycystin-2, polycystin-2-(680-796) that was proposed to contain a Ca(2+)-binding site. NMR structure determination reveals the existence of two Ca(2+)-binding sites in polycystin-2-(680-796) arranged in a typical and an atypical EF-hand motif. In the absence of Ca(2+) the protein forms a dimer that is dissociated by Ca(2+) binding. This dissociation may be related to the Ca(2+) inactivation observed earlier. The calcium affinity of the protein was determined by fluorescence and NMR spectroscopy. At 293 K, the K(D) values for the high and low affinity sites are 55 mum and 179 mum, respectively. PMID- 19546224 TI - Structural basis of the catalytic mechanism operating in open-closed conformers of lipocalin type prostaglandin D synthase. AB - Lipocalin type prostaglandin D synthase (L-PGDS) is a multifunctional protein acting as a somnogen (PGD2)-producing enzyme, an extracellular transporter of various lipophilic ligands, and an amyloid-beta chaperone in human cerebrospinal fluid. In this study, we determined the crystal structures of two different conformers of mouse L-PGDS, one with an open cavity of the beta-barrel and the other with a closed cavity due to the movement of the flexible E-F loop. The upper compartment of the central large cavity contains the catalytically essential Cys65 residue and its network of hydrogen bonds with the polar residues Ser45, Thr67, and Ser81, whereas the lower compartment is composed of hydrophobic amino acid residues that are highly conserved among other lipocalins. SH titration analysis combined with site-directed mutagenesis revealed that the Cys65 residue is activated by its interaction with Ser45 and Thr67 and that the S45A/T67A/S81A mutant showed less than 10% of the L-PGDS activity. The conformational change between the open and closed states of the cavity indicates that the mobile calyx contributes to the multiligand binding ability of L-PGDS. PMID- 19546222 TI - Epac activates the small G proteins Rap1 and Rab3A to achieve exocytosis. AB - Exocytosis of the acrosome (the acrosome reaction) relies on cAMP production, assembly of a proteinaceous fusion machinery, calcium influx from the extracellular medium, and mobilization from inositol 1,4,5-trisphosphate sensitive intracellular stores. Addition of cAMP to human sperm suspensions bypasses some of these requirements and elicits exocytosis in a protein kinase A- and extracellular calcium-independent manner. The relevant cAMP target is Epac, a guanine nucleotide exchange factor for the small GTPase Rap. We show here that a soluble adenylyl cyclase synthesizes the cAMP required for the acrosome reaction. Epac stimulates the exchange of GDP for GTP on Rap1, upstream of a phospholipase C. The Epac-selective cAMP analogue 8-pCPT-2'-O-Me-cAMP induces a phospholipase C dependent calcium mobilization in human sperm suspensions. In addition, our studies identify a novel connection between cAMP and Rab3A, a secretory granule associated protein, revealing that the latter functions downstream of soluble adenylyl cyclase/cAMP/Epac but not of Rap1. Challenging sperm with calcium or 8 pCPT-2'-O-Me-cAMP boosts the exchange of GDP for GTP on Rab3A. Recombinant Epac does not release GDP from Rab3A in vitro, suggesting that the Rab3A-GEF activation by cAMP/Epac in vivo is indirect. We propose that Epac sits at a critical point during the exocytotic cascade after which the pathway splits into two limbs, one that assembles the fusion machinery into place and another that elicits intracellular calcium release. PMID- 19546226 TI - Modulating estrogen receptor-related receptor-alpha activity inhibits cell proliferation. AB - High expression of the estrogen receptor-related receptor (ERR)-alpha in human tumors is correlated to a poor prognosis, suggesting an involvement of the receptor in cell proliferation. In this study, we show that a synthetic compound (XCT790) that modulates the activity of ERRalpha reduces the proliferation of various cell lines and blocks the G(1)/S transition of the cell cycle in an ERRalpha-dependent manner. XCT790 induces, in a p53-independent manner, the expression of the cell cycle inhibitor p21(waf/cip)(1) at the protein, mRNA, and promoter level, leading to an accumulation of hypophosphorylated Rb. Finally, XCT790 reduces cell tumorigenicity in Nude mice. PMID- 19546225 TI - Polo-like kinase 1 (PLK1) regulates interferon (IFN) induction by MAVS. AB - The mitochondria-bound adapter MAVS participates in IFN induction by recruitment of downstream partners such as members of the TRAF family, leading to activation of NF-kappaB, and the IRF3 pathways. A yeast two-hybrid search for MAVS interacting proteins yielded the Polo-box domain (PBD) of the mitotic Polo-like kinase PLK1. We showed that PBD associates with two different domains of MAVS in both dependent and independent phosphorylation events. The phosphodependent association requires the phosphopeptide binding ability of PBD. It takes place downstream of the proline-rich domain of MAVS, within an STP motif, characteristic of the binding of PLK1 to its targets, where the central Thr234 residue is phosphorylated. Its phosphoindependent association takes place at the C terminus of MAVS. PLK1 strongly inhibits the ability of MAVS to activate the IRF3 and NF-kappaB pathways and to induce IFN. Reciprocally, depletion of PLK1 can increase IFN induction in response to RIG-I/SeV or RIG-I/poly(I)-poly(C) treatments. This inhibition is dependent on the phosphoindependent association of PBD at the C terminus of MAVS where it disrupts the association of MAVS with its downstream partner TRAF3. IFN induction was strongly inhibited in cells arrested in G2/M by nocodazole, which provokes increased expression of endogenous PLK1. Interestingly, depletion of PLK1 from these nocodazole-treated cells could restore, at least partially, IFN induction. Altogether, these data demonstrate a new function for PLK1 as a regulator of IFN induction and provide the basis for the development of inhibitors preventing the PLK1/MAVS association to sustain innate immunity. PMID- 19546229 TI - Remember/Know judgments in cognitive neuroscience: An illustration of the underrepresented point of view. AB - The Remember/Know procedure is designed to easily measure two memory processes (recollection and familiarity) that are thought to serve as the basis for recognition memory decisions. This procedure is widely used in both neuroimaging studies and in studies involving amnesic patients in an effort to identify the brain structures that subserve these two memory processes. An alternative interpretation of this procedure, based on signal-detection theory, holds that Remember judgments and Know judgments are not "process pure" and are instead indicative of different degrees of memory strength (e.g., high degrees of recollection and familiarity vs. low degrees of recollection and familiarity, respectively). In the discipline of experimental psychology, the signal-detection view is widely regarded as a viable (or even preferable) alternative to the process-pure view. In the neuroscience literature, by contrast, the signal detection interpretation is rarely given serious (or even any) consideration. Because conclusions about the neuroanatomical basis of recollection and familiarity are dependent on one specific interpretation of the Remember/Know procedure, ignoring a viable alternative interpretation may be counterproductive. PMID- 19546227 TI - Genome-wide analysis of tRNA charging and activation of the eIF2 kinase Gcn2p. AB - When cells are subjected to nutritional stress, uncharged tRNAs accumulate and activate Gcn2p phosphorylation of eukaryotic initiation factor-2 (eIF2) and the general amino acid control pathway. The Gcn2p regulatory domain homologous to histidyl-tRNA synthetases is proposed to bind to uncharged tRNA, directly contributing to activation of Gcn2p. Here we apply a microarray technology to analyze genome-wide changes in tRNA charging in yeast upon activation of Gcn2p in response to amino acid starvation and high salinity, a stress not directly linked to nutritional deficiency. This microarray technology is applicable for all eukaryotic cells. Strains were starved for histidine, leucine, or tryptophan and shown to rapidly induce Gcn2p phosphorylation of eIF2. The relative charging level of all tRNAs was measured before and after starvation, and Gcn2p activation and the intracellular levels of the starved amino acid correlate with the observed decrease in tRNA charging. Interestingly, in some cases, tRNAs not charged with the starved amino acid became deacylated more rapidly than tRNAs charged with the starved amino acid. This increase in uncharged tRNA levels occurred although the intracellular levels for these non-starved amino acids remained unchanged. Additionally, treatment of a wild-type strain with high salinity stress showed transient changes in the charging of several different tRNAs. These results suggest that Gcn2p can be activated by many different tRNA species in the cell. These results also depict a complex cellular relationship between tRNA charging, amino acid availability, and non-nutrient stress. These relationships are best revealed by simultaneous monitoring of the charging level of all tRNAs. PMID- 19546228 TI - Reversibility of epithelial-mesenchymal transition (EMT) induced in breast cancer cells by activation of urokinase receptor-dependent cell signaling. AB - Hypoxia induces expression of the urokinase receptor (uPAR) and activates uPAR dependent cell signaling in cancer cells. This process promotes epithelial mesenchymal transition (EMT). uPAR overexpression in cancer cells also promotes EMT. In this study, we tested whether uPAR may be targeted to reverse cancer cell EMT. When MDA-MB 468 breast cancer cells were cultured in 1% O(2), uPAR expression increased, as anticipated. Cell-cell junctions were disrupted, vimentin expression increased, and E-cadherin was lost from cell surfaces, indicating EMT. Transferring these cells back to 21% O(2) decreased uPAR expression and reversed the signs of EMT. In uPAR-overexpressing MDA-MB 468 cells, EMT was reversed by silencing expression of endogenously produced urokinase-type plasminogen activator (uPA), which is necessary for uPAR-dependent cell signaling, or by targeting uPAR-activated cell signaling factors, including phosphatidylinositol 3-kinase, Src family kinases, and extracellular signal regulated kinase. MDA-MB 231 breast cancer cells express high levels of uPA and uPAR and demonstrate mesenchymal cell morphology under normoxic culture conditions (21% O(2)). Silencing uPA expression in MDA-MB-231 cells decreased expression of vimentin and Snail, and induced changes in morphology characteristic of epithelial cells. These results demonstrate that uPAR-initiated cell signaling may be targeted to reverse EMT in cancer. PMID- 19546230 TI - Learning-induced enhancement of feedback inhibitory synaptic transmission. AB - Olfactory-discrimination learning results with a series of intrinsic and excitatory synaptic modifications in piriform cortex pyramidal neurons. Here we show that such learning results with long-lasting enhancement of inhibitory synaptic transmission onto proximal dendrites of these pyramidal neurons. Such enhancement is mediated by a strong hyperpolarizing shift in the reversal potential of fast inhibitory postsynaptic potentials (fIPSPs). Moreover, paired pulse depression of these IPSPs, indicating enhanced GABA release, is also apparent after learning. We suggest that learning is accompanied by long-lasting enhancement of synaptic inhibition onto excitatory neurons, thus compensating for the increase of excitation in these neurons. PMID- 19546231 TI - DNA 3'-phosphatase activity is critical for rapid global rates of single-strand break repair following oxidative stress. AB - Oxidative stress is a major source of chromosome single-strand breaks (SSBs), and the repair of these lesions is retarded in neurodegenerative disease. The rate of the repair of oxidative SSBs is accelerated by XRCC1, a scaffold protein that is essential for embryonic viability and that interacts with multiple DNA repair proteins. However, the relative importance of the interactions mediated by XRCC1 during oxidative stress in vivo is unknown. We show that mutations that disrupt the XRCC1 interaction with DNA polymerase beta or DNA ligase III fail to slow SSB repair in proliferating CHO cells following oxidative stress. In contrast, mutation of the domain that interacts with polynucleotide kinase/phosphatase (PNK) and Aprataxin retards repair, and truncated XRCC1 encoding this domain fully supports this process. Importantly, the impact of mutating the protein domain in XRCC1 that binds these end-processing factors is circumvented by the overexpression of wild-type PNK but not by the overexpression of PNK harboring a mutated DNA 3'-phosphatase domain. These data suggest that DNA 3'-phosphatase activity is critical for rapid rates of chromosomal SSB repair following oxidative stress, and that the XRCC1-PNK interaction ensures that this activity is not rate limiting in vivo. PMID- 19546232 TI - Fbh1 limits Rad51-dependent recombination at blocked replication forks. AB - Controlling the loading of Rad51 onto DNA is important for governing when and how homologous recombination is used. Here we use a combination of genetic assays and indirect immunofluorescence to show that the F-box DNA helicase (Fbh1) functions in direct opposition to the Rad52 orthologue Rad22 to curb Rad51 loading onto DNA in fission yeast. Surprisingly, this activity is unnecessary for limiting spontaneous direct-repeat recombination. Instead it appears to play an important role in preventing recombination when replication forks are blocked and/or broken. When overexpressed, Fbh1 specifically reduces replication fork block induced recombination, as well as the number of Rad51 nuclear foci that are induced by replicative stress. These abilities are dependent on its DNA helicase/translocase activity, suggesting that Fbh1 exerts its control on recombination by acting as a Rad51 disruptase. In accord with this, overexpression of Fbh1 also suppresses the high levels of recombinant formation and Rad51 accumulation at a site-specific replication fork barrier in a strain lacking the Rad51 disruptase Srs2. Similarly overexpression of Srs2 suppresses replication fork block-induced gene conversion events in an fbh1Delta mutant, although an inability to suppress deletion events suggests that Fbh1 has a distinct functionality, which is not readily substituted by Srs2. PMID- 19546233 TI - Ubiquitin ligase Cbl-b is a negative regulator for insulin-like growth factor 1 signaling during muscle atrophy caused by unloading. AB - Skeletal muscle atrophy caused by unloading is characterized by both decreased responsiveness to myogenic growth factors (e.g., insulin-like growth factor 1 [IGF-1] and insulin) and increased proteolysis. Here, we show that unloading stress resulted in skeletal muscle atrophy through the induction and activation of the ubiquitin ligase Cbl-b. Upon induction, Cbl-b interacted with and degraded the IGF-1 signaling intermediate IRS-1. In turn, the loss of IRS-1 activated the FOXO3-dependent induction of atrogin-1/MAFbx, a dominant mediator of proteolysis in atrophic muscle. Cbl-b-deficient mice were resistant to unloading-induced atrophy and the loss of muscle function. Furthermore, a pentapeptide mimetic of tyrosine(608)-phosphorylated IRS-1 inhibited Cbl-b-mediated IRS-1 ubiquitination and strongly decreased the Cbl-b-mediated induction of atrogin-1/MAFbx. Our results indicate that the Cbl-b-dependent destruction of IRS-1 is a critical dual mediator of both increased protein degradation and reduced protein synthesis observed in unloading-induced muscle atrophy. The inhibition of Cbl-b-mediated ubiquitination may be a new therapeutic strategy for unloading-mediated muscle atrophy. PMID- 19546234 TI - Interaction of FLASH with arsenite resistance protein 2 is involved in cell cycle progression at S phase. AB - FLASH has been shown to be required for S phase progression and to interact with a nuclear protein, ataxia-telangiectasia locus (NPAT), a component of Cajal bodies in the nucleus and an activator of histone transcription. We investigated the role of human FLASH by using an inducible FLASH knockdown system in the presence or absence of various mutant forms of mouse FLASH. While carboxyl terminal deletion mutants of FLASH, which do not interact with NPAT, can support S phase progression, its amino-terminal deletion mutants, which are unable to self associate, cannot support S phase progression, replication-dependent histone transcription, or the formation of Cajal bodies. Furthermore, FLASH was shown to be associated with arsenite resistance protein 2 (ARS2) through its central region, which is composed of only 13 amino acids. The expression of ARS2 and the interaction between FLASH and ARS2 are required for S phase progression. Taking these results together, FLASH functions in S phase progression through interaction with ARS2. PMID- 19546235 TI - A randomly integrated transgenic H19 imprinting control region acquires methylation imprinting independently of its establishment in germ cells. AB - The imprinted expression of the mouse Igf2/H19 locus is governed by the differential methylation of the imprinting control region (ICR), which is established initially in germ cells and subsequently maintained in somatic cells, depending on its parental origin. By grafting a 2.9-kbp H19 ICR fragment into a human beta-globin yeast artificial chromosome in transgenic mice, we previously showed that the ICR could recapitulate imprinted methylation and expression at a heterologous locus, suggesting that the H19 ICR in the beta-globin locus contained sufficient information to maintain the methylation mark (K. Tanimoto, M. Shimotsuma, H. Matsuzaki, A. Omori, J. Bungert, J. D. Engel, and A. Fukamizu, Proc. Natl. Acad. Sci. USA 102:10250-10255, 2005). Curiously, however, the transgenic H19 ICR was not methylated in sperm, which was distinct from that seen in the endogenous locus. Here, we reevaluated the ability of the H19 ICR to mark the parental origin using more rigid criteria. In the testis, the methylation levels of the solitary 2.9-kbp transgenic ICR fragment varied significantly between six transgenic mouse lines. However, in somatic cells, the paternally inherited ICR fragment exhibited consistently higher methylation levels at five out of six randomly integrated sites in the mouse genome. These results clearly demonstrated that the H19 ICR could acquire parent-of-origin-dependent methylation after fertilization independently of the chromosomal integration site or the prerequisite methylation acquisition in male germ cells. PMID- 19546236 TI - Hypoxia-inducible factor 1alpha induces fibrosis and insulin resistance in white adipose tissue. AB - Adipose tissue can undergo rapid expansion during times of excess caloric intake. Like a rapidly expanding tumor mass, obese adipose tissue becomes hypoxic due to the inability of the vasculature to keep pace with tissue growth. Consequently, during the early stages of obesity, hypoxic conditions cause an increase in the level of hypoxia-inducible factor 1alpha (HIF1alpha) expression. Using a transgenic model of overexpression of a constitutively active form of HIF1alpha, we determined that HIF1alpha fails to induce the expected proangiogenic response. In contrast, we observed that HIF1alpha initiates adipose tissue fibrosis, with an associated increase in local inflammation. "Trichrome- and picrosirius red positive streaks," enriched in fibrillar collagens, are a hallmark of adipose tissue suffering from the early stages of hypoxia-induced fibrosis. Lysyl oxidase (LOX) is a transcriptional target of HIF1alpha and acts by cross-linking collagen I and III to form the fibrillar collagen fibers. Inhibition of LOX activity by beta-aminoproprionitrile treatment results in a significant improvement in several metabolic parameters and further reduces local adipose tissue inflammation. Collectively, our observations are consistent with a model in which adipose tissue hypoxia serves as an early upstream initiator for adipose tissue dysfunction by inducing a local state of fibrosis. PMID- 19546238 TI - Identification and profiling of circulating metabolites of atazanavir, a HIV protease inhibitor. AB - Atazanavir is a commonly prescribed protease inhibitor for treatment of HIV-1 infection. Thus far, only limited data are available on the in vivo metabolism of the drug. Three systemic circulating metabolites have been reported, but their chemical structures have not been released publicly. Atazanavir metabolites may contribute to its effectiveness but also to its toxicity and interactions. Thus, there is a need for extensive metabolic profiling of atazanavir. Our goals were to screen and identify previously unknown atazanavir metabolites and to develop a sensitive metabolite profiling method in plasma. Five atazanavir metabolites were detected and identified in patient samples using liquid chromatography coupled to linear ion trap mass spectrometry: one N-dealkylation product (M1), two metabolites resulting from carbamate hydrolysis (M2 and M3), a hydroxylated product (M4), and a keto-metabolite (M5). For sensitive semiquantitative analysis of the metabolites in plasma, the method was transferred to liquid chromatography coupled to triple quadrupole mass spectrometry. In 12 patient samples, all the metabolites could be detected, and possible other potential atazanavir keto metabolites were found. Atazanavir metabolite levels were positively correlated with atazanavir levels, but interindividual variability was high. The developed atazanavir metabolic screening method can now be used for further clinical pharmacological research with this antiretroviral agent. PMID- 19546239 TI - Modulation of the partition coefficient between octanol and buffer at pH 7.4 and pKa to achieve the optimum balance of blood clearance and volume of distribution for a series of tetrahydropyran histamine type 3 receptor antagonists. AB - The relationship between rat pharmacokinetics and physicochemical parameters [the partition coefficient between octanol and buffer at pH 7.4 (log D((7.4))) and pK(a)] was studied for a series of tetrahydropyran compounds. Sixteen compounds ranging in log D((7.4)) 0.1 to 1.8 were administered intravenously to rats, and the pharmacokinetic parameters were determined from blood concentration time curves. Across the series, a weak correlation was observed between log D((7.4)) and blood clearance, suggesting that log D((7.4)) values less than 0.5 were required to prevent clearance at hepatic blood flow. In terms of the volume of distribution (V(d)), the compounds fell into three distinct subseries characterized by the number of basic centers and differences in ionization of each basic center at physiological pH. These were referred to as the monobasic, weak second base, and strong second base subseries. All the compounds exhibited V(d) greater than body water, as would be expected from their lipophilic and basic nature. For a given clog P, the strong second base subseries showed higher V(d) than the weak second base subseries, which in turn exhibited higher values than the monobasic subseries. In addition, for the weak second base subseries, V(d) could be tuned by modulating the pK(a) of the second basic center. This relationship was rationalized in respect to the interactions of the ionizable centers with phospholipid heads in the cell membrane and/or lysosomal trapping. Compounds in the weak second base subseries showed optimal V(d), and when combined with a log D((7.4)) of 0.1, driving to moderate blood clearance, one compound showed the optimal pharmacokinetic profile. PMID- 19546237 TI - TOR complex 2 controls gene silencing, telomere length maintenance, and survival under DNA-damaging conditions. AB - The Target Of Rapamycin (TOR) kinase belongs to the highly conserved eukaryotic family of phosphatidylinositol-3-kinase-related kinases (PIKKs). TOR proteins are found at the core of two distinct evolutionarily conserved complexes, TORC1 and TORC2. Disruption of TORC1 or TORC2 results in characteristically dissimilar phenotypes. TORC1 is a major cell growth regulator, while the cellular roles of TORC2 are not well understood. In the fission yeast Schizosaccharomyces pombe, Tor1 is a component of the TORC2 complex, which is particularly required during starvation and various stress conditions. Our genome-wide gene expression analysis of Deltator1 mutants indicates an extensive similarity with chromatin structure mutants. Consistently, TORC2 regulates several chromatin-mediated functions, including gene silencing, telomere length maintenance, and tolerance to DNA damage. These novel cellular roles of TORC2 are rapamycin insensitive. Cells lacking Tor1 are highly sensitive to the DNA-damaging drugs hydroxyurea (HU) and methyl methanesulfonate, similar to mutants of the checkpoint kinase Rad3 (ATR). Unlike Rad3, Tor1 is not required for the cell cycle arrest in the presence of damaged DNA. Instead, Tor1 becomes essential for dephosphorylation and reactivation of the cyclin-dependent kinase Cdc2, thus allowing reentry into mitosis following recovery from DNA replication arrest. Taken together, our data highlight critical roles for TORC2 in chromatin metabolism and in promoting mitotic entry, most notably after recovery from DNA-damaging conditions. These data place TOR proteins in line with other PIKK members, such as ATM and ATR, as guardians of genome stability. PMID- 19546240 TI - Up-regulation of UDP-glucuronosyltransferase (UGT) 1A4 by 17beta-estradiol: a potential mechanism of increased lamotrigine elimination in pregnancy. AB - Oral clearance of lamotrigine, an antiepileptic drug commonly used in pregnant women, is increased in pregnancy by unknown mechanisms. In this study, we show that 17beta-estradiol (E(2)) up-regulates expression of UDP glucuronosyltransferase (UGT) 1A4, the major enzyme responsible for elimination of lamotrigine. Endogenous mRNA expression levels of UGT1A4 in estrogen receptor (ER) alpha-negative HepG2 cells were induced 2.3-fold by E(2) treatment in the presence of ER alpha expression. E(2) enhanced transcriptional activity of UGT1A4 in a concentration-dependent manner in HepG2 cells when ER alpha was cotransfected. Induction of UGT1A4 transcriptional activity by E(2) was also observed in ER alpha-positive MCF7 cells, which was abrogated by pretreatment with the antiestrogen fulvestrant (ICI 182,780). Analysis of UGT1A4 upstream regions using luciferase reporter assays identified a putative specificity protein-1 (Sp1) binding site (-1906 to -1901 base pairs) that is critical for the induction of UGT1A4 transcriptional activity by E(2). Deletion of the Sp1 binding sequence abolished the UGT1A4 up-regulation by E(2), and Sp1 bound to the putative Sp1 binding site as determined by a electrophoretic mobility shift assay. Analysis of ER alpha domains using ER alpha mutants revealed that the activation function (AF) 1 and AF2 domains but not the DNA binding domain of ER alpha are required for UGT1A4 induction by E(2) in HepG2 cells. Finally, E(2) treatment increased lamotrigine glucuronidation in ER alpha-transfected HepG2 cells. Together, our data indicate that up-regulation of UGT1A4 expression by E(2) is mediated by both ER alpha and Sp1 and is a potential mechanism contributing to the enhanced elimination of lamotrigine in pregnancy. PMID- 19546241 TI - Microcephalin and pericentrin regulate mitotic entry via centrosome-associated Chk1. AB - Primary microcephaly, Seckel syndrome, and microcephalic osteodysplastic primordial dwarfism type II (MOPD II) are disorders exhibiting marked microcephaly, with small brain sizes reflecting reduced neuron production during fetal life. Although primary microcephaly can be caused by mutations in microcephalin (MCPH1), cells from patients with Seckel syndrome and MOPD II harbor mutations in ataxia telangiectasia and Rad3 related (ATR) or pericentrin (PCNT), leading to disturbed ATR signaling. In this study, we show that a lack of MCPH1 or PCNT results in a loss of Chk1 from centrosomes with subsequently deregulated activation of centrosomal cyclin B-Cdk1. PMID- 19546242 TI - MURC/Cavin-4 and cavin family members form tissue-specific caveolar complexes. AB - Polymerase I and transcript release factor (PTRF)/Cavin is a cytoplasmic protein whose expression is obligatory for caveola formation. Using biochemistry and fluorescence resonance energy transfer-based approaches, we now show that a family of related proteins, PTRF/Cavin-1, serum deprivation response (SDR)/Cavin 2, SDR-related gene product that binds to C kinase (SRBC)/Cavin-3, and muscle restricted coiled-coil protein (MURC)/Cavin-4, forms a multiprotein complex that associates with caveolae. This complex can constitutively assemble in the cytosol and associate with caveolin at plasma membrane caveolae. Cavin-1, but not other cavins, can induce caveola formation in a heterologous system and is required for the recruitment of the cavin complex to caveolae. The tissue-restricted expression of cavins suggests that caveolae may perform tissue-specific functions regulated by the composition of the cavin complex. Cavin-4 is expressed predominantly in muscle, and its distribution is perturbed in human muscle disease associated with Caveolin-3 dysfunction, identifying Cavin-4 as a novel muscle disease candidate caveolar protein. PMID- 19546243 TI - Desmoglein 1-dependent suppression of EGFR signaling promotes epidermal differentiation and morphogenesis. AB - Dsg1 (desmoglein 1) is a member of the cadherin family of Ca(2+)-dependent cell adhesion molecules that is first expressed in the epidermis as keratinocytes transit out of the basal layer and becomes concentrated in the uppermost cell layers of this stratified epithelium. In this study, we show that Dsg1 is not only required for maintaining epidermal tissue integrity in the superficial layers but also supports keratinocyte differentiation and suprabasal morphogenesis. Dsg1 lacking N-terminal ectodomain residues required for adhesion remained capable of promoting keratinocyte differentiation. Moreover, this capability did not depend on cytodomain interactions with the armadillo protein plakoglobin or coexpression of its companion suprabasal cadherin, Dsc1 (desmocollin 1). Instead, Dsg1 was required for suppression of epidermal growth factor receptor-Erk1/2 (extracellular signal-regulated kinase 1/2) signaling, thereby facilitating keratinocyte progression through a terminal differentiation program. In addition to serving as a rigid anchor between adjacent cells, this study implicates desmosomal cadherins as key components of a signaling axis governing epithelial morphogenesis. PMID- 19546244 TI - The Drosophila deoxyhypusine hydroxylase homologue nero and its target eIF5A are required for cell growth and the regulation of autophagy. AB - Hypusination is a unique posttranslational modification by which lysine is transformed into the atypical amino acid hypusine. eIF5A (eukaryotic initiation factor 5A) is the only known protein to contain hypusine. In this study, we describe the identification and characterization of nero, the Drosophila melanogaster deoxyhypusine hydroxylase (DOHH) homologue. nero mutations affect cell and organ size, bromodeoxyuridine incorporation, and autophagy. Knockdown of the hypusination target eIF5A via RNA interference causes phenotypes similar to nero mutations. However, loss of nero appears to cause milder phenotypes than loss of eIF5A. This is partially explained through a potential compensatory mechanism by which nero mutant cells up-regulate eIF5A levels. The failure of eIF5A up-regulation to rescue nero mutant phenotypes suggests that hypusination is required for eIF5A function. Furthermore, expression of enzymatically impaired forms of DOHH fails to rescue nero clones, indicating that hypusination activity is important for nero function. Our data also indicate that nero and eIF5A are required for cell growth and affect autophagy and protein synthesis. PMID- 19546246 TI - Profound CD4+/CCR5+ T cell expansion is induced by CD8+ lymphocyte depletion but does not account for accelerated SIV pathogenesis. AB - Depletion of CD8(+) lymphocytes during acute simian immunodeficiency virus (SIV) infection of rhesus macaques (RMs) results in irreversible prolongation of peak level viral replication and rapid disease progression, consistent with a major role for CD8(+) lymphocytes in determining postacute-phase viral replication set points. However, we report that CD8(+) lymphocyte depletion is also associated with a dramatic induction of proliferation among CD4(+) effector memory T (T(EM)) cells and, to a lesser extent, transitional memory T (T(TrM)) cells, raising the question of whether an increased availability of optimal (activated/proliferating), CD4(+)/CCR5(+) SIV "target" cells contributes to this accelerated pathogenesis. In keeping with this, depletion of CD8(+) lymphocytes in SIV(-) RMs led to a sustained increase in the number of potential CD4(+) SIV targets, whereas such depletion in acute SIV infection led to increased target cell consumption. However, we found that the excess CD4(+) T(EM) cell proliferation of CD8(+) lymphocyte-depleted, acutely SIV-infected RMs was completely inhibited by interleukin (IL)-15 neutralization, and that this inhibition did not abrogate the rapidly progressive infection in these RMs. Moreover, although administration of IL-15 during acute infection induced robust CD4(+) T(EM) and T(TrM) cell proliferation, it did not recapitulate the viral dynamics of CD8(+) lymphocyte depletion. These data suggest that CD8(+) lymphocyte function has a larger impact on the outcome of acute SIV infection than the number and/or activation status of target cells available for infection and viral production. PMID- 19546247 TI - Cardiac 12/15 lipoxygenase-induced inflammation is involved in heart failure. AB - To identify a novel target for the treatment of heart failure, we examined gene expression in the failing heart. Among the genes analyzed, Alox15 encoding the protein 12/15 lipoxygenase (LOX) was markedly up-regulated in heart failure. To determine whether increased expression of 12/15-LOX causes heart failure, we established transgenic mice that overexpressed 12/15-LOX in cardiomyocytes. Echocardiography showed that Alox15 transgenic mice developed systolic dysfunction. Cardiac fibrosis increased in Alox15 transgenic mice with advancing age and was associated with the infiltration of macrophages. Consistent with these observations, cardiac expression of monocyte chemoattractant protein 1 (MCP 1) was up-regulated in Alox15 transgenic mice compared with wild-type mice. Treatment with 12-hydroxy-eicosatetraenoic acid, a major metabolite of 12/15-LOX, increased MCP-1 expression in cardiac fibroblasts and endothelial cells but not in cardiomyocytes. Inhibition of MCP-1 reduced the infiltration of macrophages into the myocardium and prevented both systolic dysfunction and cardiac fibrosis in Alox15 transgenic mice. Likewise, disruption of 12/15-LOX significantly reduced cardiac MCP-1 expression and macrophage infiltration, thereby improving systolic dysfunction induced by chronic pressure overload. Our results suggest that cardiac 12/15-LOX is involved in the development of heart failure and that inhibition of 12/15-LOX could be a novel treatment for this condition. PMID- 19546249 TI - Cytokine-dependent regulation of NADPH oxidase activity and the consequences for activated T cell homeostasis. AB - Cellular dependence on growth factors for survival is developmentally programmed and continues in adult metazoans. Antigen-activated T cell apoptosis in the waning phase of the immune response is thought to be triggered by depletion of cytokines from the microenvironment. T cell apoptosis resulting from cytokine deprivation is mediated by reactive oxygen species (ROS), but their source and position in the apoptotic cascade is poorly understood. RNA interference approaches implicated the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in neglect-induced apoptosis in T cells. Using mice deficient for the catalytic subunit gp91(phox) to characterize the molecular link to activated T cell apoptosis, we show that gp91(phox)-deficient T (T(-/-)) cells generated mitochondrial superoxide but had diminished hydrogen peroxide production in response to neglect, which, in turn, regulated Jun N-terminal kinase-dependent Bax activation and apoptosis. Activated T(-/-) cells were distinguished by improved survival after activation by superantigens in vivo, adoptive transfers into congenic hosts, and higher recall responses after immunization. Thus, the NADPH oxidase may regulate adaptive immunity in addition to its previously well characterized role in the innate response. PMID- 19546251 TI - Pharmacological aspects and potential new clinical applications of ketamine: reevaluation of an old drug. AB - Ketamine, the phencyclidine derivative described in 1965, is an intravenous anesthetic with a variety of applications. The enthusiasm following its initial release subsided due to side effects from the central nervous system. New anesthetics limited the role of ketamine in anesthetic practice. However, its hemodynamically stable profile, along with its beneficial respiratory properties and analgesic potency, rendered the drug invaluable in battlefield medicine, sedation of the uncooperative child, analgesia, and sedation in burn units. Reevaluation, though, of analgesic properties of ketamine resulted in new interest regarding its use in perioperative and chronic pain management. Moreover, recent studies in the effects of the substance on intracranial pressure and cerebral blood flow led to revising the recommendation against its use in brain injury. Furthermore, the bronchodilating effects of the substance led to increasing interest for potential use in asthma treatment. In addition, separation of the 2 enantiomers and subsequent separate studies indicated beneficial results of the S(+) one. Thus, new controlled multicentered clinical trials are to be conducted to justify approval for new uses of ketamine and take advantage of its unique range of applications. PMID- 19546248 TI - T-bet is essential for encephalitogenicity of both Th1 and Th17 cells. AB - The extent to which myelin-specific Th1 and Th17 cells contribute to the pathogenesis of experimental autoimmune encephalomyelitis (EAE) is controversial. Combinations of interleukin (IL)-1beta, IL-6, and IL-23 with transforming growth factor beta were used to differentiate myelin-specific T cell receptor transgenic T cells into Th17 cells, none of which could induce EAE, whereas Th1 cells consistently transferred disease. However, IL-6 was found to promote the differentiation of encephalitogenic Th17 cells. Further analysis of myelin specific T cells that were encephalitogenic in spontaneous EAE and actively induced EAE demonstrated that T-bet expression was critical for pathogenicity, regardless of cytokine expression by the encephalitogenic T cells. These data suggest that encephalitogenicity of myelin-specific T cells appears to be mediated by a pathway dependent on T-bet and not necessarily pathway-specific end products, such as interferon gamma and IL-17. PMID- 19546250 TI - Population pharmacokinetic analyses to evaluate the influence of intrinsic and extrinsic factors on exposure of prasugrel active metabolite in TRITON-TIMI 38. AB - Serial pharmacokinetic (PK) sampling in 1159 patients from TRITON-TIMI 38 was undertaken. A multilinear regression model was used to quantitatively predict prasugrel's active metabolite (Pras-AM) concentrations from its 2 downstream inactive metabolites. Population-based methods were then applied to Pras-AM concentration data to characterize the PK. The potential influence of body weight, body mass index, age, sex, renal function, diabetes, tobacco use, and other disease status on Bayesian estimates of Pras-AM exposures was assessed. The PK of Pras-AM was adequately described by a multicompartmental model and consistent with results from previous studies. The systemic exposure of prasugrel was not appreciably affected by body mass index, gender, diabetes, smoking, and renal impairment. Pras-AM mean exposure in patients weighing <60 kg (4.1%) was 30% (90% confidence interval [CI] 1.16-1.45) higher than exposure in patients > or =60 kg. Mean Pras-AM exposures for patients > or =75 years (10.5%) were 19% (90% CI: 1.11-1.28) higher compared with patients <75 years. PMID- 19546252 TI - Influence of glomerular filtration rate on the pharmacokinetics of cyclophosphamide enantiomers in patients with lupus nephritis. AB - The pharmacokinetics of cyclophosphamide (CYC) enantiomers were evaluated in patients with lupus nephritis distributed in 2 groups according to creatinine clearance: group 1 (90.6-144.6 mL/min/1.73 m(2)) and group 2 (42.8-76.4 mL/min/1.73 m(2)). All patients were treated with 0.75 to 1.3 g of racemic CYC as a 2-hour infusion and with 1 mg intravenous midazolam as a drug-metabolizing marker. CYC enantiomers and midazolam concentrations in plasma were measured by liquid chromatography/tandem mass spectrometry (LC/MS/MS). The following differences (Wilcoxon test, P < or = .05) were observed between the (S)-(-) and (R)-(+) enantiomers: AUC(0-infinity) 152.41 vs 129.25 microg.h/mL, CL 3.28 vs 3.89 L/h, Vd 31.38 vs 29.74 L, and t((1/2)) 6.79 vs 5.56 h for group 1 and AUC(0 infinity) 167.20 vs 139.08 microg.h/mL, CL 2.99 vs 3.59 L/h, and t((1/2)) 6.15 vs 4.99 h for group 2. No differences (Mann test, P < or = .05) were observed between groups 1 and 2 in the pharmacokinetic parameters of both enantiomers. No significant relationship was observed between midazolam clearance (2.92-16.40 mL/min.kg) and clearance of each CYC enantiomer. In conclusion, CYC kinetic disposition is enantioselective, resulting in higher exposures of the (S)-(-) enantiomer in lupus nephritis patients, and the pharmacokinetic parameters of both enantiomers are not altered by the worsening of renal condition. PMID- 19546253 TI - Structured re-assessment system at 6 months after a disabling stroke: a randomised controlled trial with resource use and cost study. AB - BACKGROUND: national policy recommends routine re-assessment of disabled patients and their carers at 6 months after stroke onset. The clinical and resource outcomes of this policy were investigated. DESIGN: prospective, single-blind, randomised controlled trial in two centres. PARTICIPANTS: a total of 265 patients with a disabling stroke and their carers. INTERVENTIONS: a structured re assessment system for patients and their carers at 6 months post-stroke or existing care. OUTCOME MEASURES: primary: patient independence (Frenchay activities index) and carer stress (general health questionnaire 28). Secondary: activities of daily living, mood state, satisfaction with services, carer strain index, health and social service resource use and costs. RESULTS: independence at 12 months post-stroke was similar in both groups (Frenchay activities index, adjusted mean difference 0.64; 95% confidence interval -0.74-2.02). Emotional distress in carers was similar in both groups (general health questionnaire 28, mean difference 0.02; 95% confidence interval -0.95-1.00). Results for the secondary outcome measures and total mean costs were similar for both groups. The intervention group patients used 301 fewer hospital bed days and 1,631 fewer care home bed days. CONCLUSIONS: the structured, systematic re-assessment for patients and their carers was not associated with any clinically significant evidence of benefit at 12 months. Health and social care resource use and mean cost per patient were broadly similar in both groups. TRIAL REGISTRATION: International Standard Randomised Controlled Trial Register; number: ISRCTN55412871. PMID- 19546254 TI - 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces calcium influx through T-type calcium channel and enhances lysosomal exocytosis and insulin secretion in INS-1 cells. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) has been associated with diabetes in several epidemiological studies. However, the diabetogenic action of TCDD on pancreatic cells is unclear. Here, we investigated the direct toxic effects of TCDD on a rat insulin-secreting beta cell line. We found that TCDD enhances exocytosis of MTT formazan and lysosomal proteins such as beta-hexosaminindase and Lamp-1. This TCDD-induced exocytosis was abrogated by T-type calcium channel blockers (mibefradil, flunarizine) but not by an aryl hydrocarbon receptor antagonist (alpha-naphtoflavone). Indeed, cytosolic calcium levels were increased by TCDD. Furthermore, TCDD stimulated insulin secretion, which was inhibited by flunarizine. Taken together, our results suggest that TCDD-induced calcium influx via T-type channels regulates vesicular trafficking, such as lysosomal and secretory granule exocytosis, and that TCDD might exert adverse effects on beta cells by continuous insulin release followed by beta cell exhaustion. This could contribute to the link between TCDD exposure and the risk of developing diabetes. PMID- 19546255 TI - Development of a high-throughput human HepG(2) dual luciferase assay for detection of metabolically activated hepatotoxicants and genotoxicants. AB - Hepatic toxicity remains a major concern for drug failure; therefore, a thorough examination of chemically induced liver toxicity is essential for a robust safety evaluation. Current hypotheses suggest that the metabolic activation of a drug to a reactive intermediate is an important process. In this article, we describe a new high-throughput GADD45beta reporter assay developed for assessing potential liver toxicity. Most importantly, this assay utilizes a human cell line and incorporates metabolic activation and thus provides significant advantage over other comparable assays used to determine hepatotoxicity. Our assay has low compound requirement and relies upon 2 reporter genes cotransfected into the HepG(2) cells. The gene encoding Renilla luciferase is fused to the CMV promoter and provides a control for cell numbers. The firefly luciferase gene is fused to the GADD45beta promoter and used to report an increase in DNA damage. A dual luciferase assay is performed by measuring the firefly and Renilla luciferase activities in the same sample. Results are expressed as the ratio of the 2 luciferase activities; increases over the control are interpreted as evidence of stress responses. This mammalian dual luciferase reporter has been characterized with, and without, metabolic activation using positive and negative control agents. Our data demonstrate that this assay provides for an assessment of potential toxic metabolites, is adaptable to a high-throughput platform, and yields data that accurately and reproducibly detect hepatotoxicants. PMID- 19546256 TI - Heat shock protein 72 protects kidney proximal tubule cells from injury induced by triptolide by means of activation of the MEK/ERK pathway. AB - Triptolide, which has been used to treat inflammatory diseases, has also been reported to inhibit proliferation of cancer cells. However, it can cause severe nephrotoxicity, limiting its clinical use. Here, nephrotoxicity of triptolide was observed in vivo and in vitro. Heat shock protein 72 (HSP72) was upregulated during kidney injury in rats. HSP72 partially protected human kidney proximal tubule cell lines HK-2 and HKC from triptolide-induced injury. Phospho-Raf, phospho-MEK and phospho-ERK were elevated in HK-2 cells that overexpressed HSP72 after either heat shock or triptolide treatment, and downregulated when HSP72 was repressed by siRNA. The participation of the MEK/ERK1/2 pathway was confirmed by exposure of the cells to the MEK inhibitor U0126. Collectively, our results suggested that HSP72 plays a protective role by means of the MEK/ERK pathway, against triptolide-induced kidney injury. PMID- 19546257 TI - Calcium and calmodulin regulate mercury-induced phospholipase D activation in vascular endothelial cells. AB - Earlier, we reported that mercury, the environmental risk factor for cardiovascular diseases, activates vascular endothelial cell (EC) phospholipase D (PLD). Here, we report the novel and significant finding that calcium and calmodulin regulated mercury-induced PLD activation in bovine pulmonary artery ECs (BPAECs). Mercury (mercury chloride, 25 microM; thimerosal, 25 microM; methylmercury, 10 microM) significantly activated PLD in BPAECs. Calcium chelating agents and calcium depletion of the medium completely attenuated the mercury-induced PLD activation in ECs. Calmodulin inhibitors significantly attenuated mercury-induced PLD activation in BPAECs. Despite the absence of L type calcium channels in ECs, nifedipine, nimodipine, and diltiazem significantly attenuated mercury-induced PLD activation and cytotoxicity in BPAECs. This study demonstrated the importance of calcium and calmodulin in the regulation of mercury-induced PLD activation and the protective action of L-type calcium channel blockers against mercury cytotoxicity in vascular ECs, suggesting mechanisms of mercury vasculotoxicity and mercury-induced cardiovascular diseases. PMID- 19546258 TI - Cellular mechanism of the QT prolongation induced by sulpiride. AB - In this study, the authors investigated the electrophysiological effect of sulpiride on cardiac repolarization using conventional microelectrode recording techniques in isolated canine Purkinje fibers and a whole-cell patch clamp technique in transiently transfected cells with the hERG, KCNQ1/KCNE1, KCNJ2, and SCN5A cDNA and in rat cardiac myocytes for I(Ca). In studies of action potential duration, 10 microM, 100 microM, 300 microM, and 1 mM sulpiride prolonged action potential duration in a concentration-dependent manner. In studies of cardiac ion channels, sulpiride did not significantly affect I(Na), I(Ca), I(Ks), I(K1), except for I(Kr). Sulpiride dose-dependently decreased the hERG tail current. It is considered that the prolonged action potential duration by sulpiride was mainly the result of inhibition of the hERG channel. The data suggest that the clinical use of sulpiride is reasonable within therapeutic plasma concentrations, but all patients taking this drug should be cautiously monitored for clinical signs of long-QT syndrome and severe arrhythmia. PMID- 19546259 TI - Evaluation of the developmental toxicity of 1-(1,2,3,4,5,6,7,8-octahydro-2,3,8,8 tetramethyl-2-naphthalenyl) ethanone (OTNE) in rats. AB - The developmental toxicity of 1-(1,2,3,4,5,6,7,8-Octahydro-2,3,8,8-tetramethyl-2 naphthalenyl) ethanone (OTNE), a widely used fragrance ingredient, was evaluated in pregnant Sprague-Dawley rats (25/group) gavaged with dosages of 0 (water), 96, 240, or 480 mg/kg/d on days 7 through 17 of gestation (GDs 7-17). Rats were observed for clinical signs, abortions, premature deliveries, body weights, and feed intake. Caesarean section and necropsy were performed on GD 21. Fetuses were weighed and examined for gender, gross external changes, and soft tissue or skeletal alterations. No deaths or premature deliveries were attributed to OTNE. OTNE-related clinical signs included significantly increased incidences of excessive salivation in all 3 treatment groups, and urine-stained abdominal fur in the high dosage group. Mean body weight gains were significantly reduced by all OTNE dosages on GDs 7-10, while at 480 mg/kg/d, significant reductions continued through the remainder of the dosage period. Feed consumption generally paralleled body weight gains. Fetal body weights were reduced by 480 mg/kg/d, but not to a statistically significant degree. No fetal gross external, soft tissue, or skeletal malformations or variations were attributable to OTNE. Based on these data, maternal and developmental no-observable-adverse-effect-levels (NOAELs) of 240 mg/kg/d were established for OTNE. It was concluded that OTNE is not a developmental toxicant in rats under the conditions of this study, and that a margin of safety greater than 2700 exists between reversible developmental delays in rats and the calculated daily human exposure level of 0.086 mg/kg/d. PMID- 19546260 TI - Acute toxicity evaluation of proliferol: a dose-escalating, placebo-controlled study in swine. AB - Prolotherapy is one of the many treatments available for chronic musculoskeletal disorders. A commonly used drug contains dextrose 12.5%, glycerin 12.5%, phenol 1.0%, and lidocaine hydrochloride 0.25% in aqueous solution (recently termed Proliferol). For chronic low back pain, this is injected into lumbosacral ligaments to stimulate connective tissue repair. Despite generally positive clinical results, the toxicity of this drug is not well characterized and was assessed in 48 (24 male, 24 female) Yucatan miniature swine randomly assigned to low (1x), medium (5x), or high (10x) dose or saline placebo. Outcomes included clinical observations, clinical chemistry, hematology, coagulation, urinalysis, toxicokinetics, and full gross and microscopic histopathology after 24 hours or 14 days. Findings attributable to Proliferol after 24 hours included dose response elevations in alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and creatine kinase, which returned to normal after 14 days. There were no remarkable findings in hematology, coagulation, or urinalysis. Urine concentrations of lidocaine and phenol both peaked after 8 hours. Histopathology findings after 24 hours included hemorrhage, inflammation, necrosis, and vascular changes in the ligaments and adjacent soft tissues at the sites of injection. After 14 days, there was evidence of repair under way, with fibrosis and skeletal muscle regeneration at the injection sites. PMID- 19546261 TI - Alternative strategies for toxicity testing of species-specific biopharmaceuticals. AB - Although toxicology studies should always be conducted in pharmacologically relevant species, the specificity of many biopharmaceuticals can present challenges in identification of a relevant species. In certain cases, that is, when the clinical product is active only in humans or chimpanzees, or if the clinical candidate is active in other species but immunogenicity limits the ability to conduct a thorough safety assessment, alternative approaches to evaluating the safety of a biopharmaceutical must be considered. Alternative approaches, including animal models of disease, genetically modified mice, or use of surrogate molecules, may improve the predictive value of preclinical safety assessments of species-specific biopharmaceuticals, although many caveats associated with these models must be considered. Because of the many caveats that are discussed in this article, alternative approaches should only be used to evaluate safety when the clinical candidate cannot be readily tested in at least one relevant species to identify potential hazards. PMID- 19546265 TI - Improving post-hypoglycaemic patient safety in the prehospital environment: a systematic review. AB - To determine the extent to which post-hypoglycaemic patients with diabetes, who are prescribed oral hypoglycaemic agents (OHA) are at risk of repeat hypoglycaemic events (RHE) after being treated in the prehospital environment and whether they should be transported to hospital regardless of their post-treatment response, a systematic literature review was carried out using an overlapping retrieval strategy that included both published and unpublished literature. Retrieved papers were reviewed by each author for inclusion. Disagreements regarding inclusion were resolved through discussion. Ninety-eight papers and other relevant material were retrieved using the developed search strategy. Twenty-three papers and other relevant material were included in the final review. A narrative synthesis of the findings is presented. Although several case reports demonstrate the risks associated with repeat or prolonged hypoglycaemia, the review was unable to locate any specific high quality research in this area. Consequently, caution is required in interpreting the findings of the studies. Post-hypoglycaemic patients treated in the prehospital environment have a 2-7% risk of experiencing a RHE within 48 h. The literature retrieved in this study recognises the potential for OHA to cause RHE. However, the extent to which this occurs in practice remains unknown. This lack of evidence has led to the recommendation that conservative management, through admission to hospital, is appropriate. The review concludes with recommendations for both practice and research. PMID- 19546266 TI - Effect of a pathway bundle on length of stay. AB - BACKGROUND: Pathways to guide clinical care are well accepted and used in many emergency departments. We wanted to introduce a number ("bundle") over a short space of time and involve the whole patient stay in the pathway. It was hypothesised that a more efficient process would result with an overall reduction in length of stay (LoS). METHODS: A "bundle" of 14 evidence-based pathways of care was introduced into a medium-sized district general hospital (DGH) in late 2006/early 2007. These pathways covered emergency department care and acute medical care for a period of up to 48 h. A total of 8184 acute emergency admission episodes were audited, 3852 in the 8 months before introduction of the new pathways and 4332 in the 8 months after their introduction. RESULTS: The overall effect of introducing the pathway bundle had a trend towards reduction in LoS by 0.2 days (95% CI -0.2 to 0.5), but this was not statistically significant (p>0.1). However, in those patients with T and -164T>C result in lower transcription of MTTP in vivo in the heart, liver, and macrophages. In vitro experiments indicated that the minor -164C allele mediates the lower gene expression and that C/EBP binds to the polymorphic region in an allele-specific manner. Furthermore, homozygous carriers of the -164C were found to have increased risk for IHD as shown in a case-control study including a total of 544 IHD patients and 544 healthy control subjects. We concluded that carriers of the minor -164C allele have lower expression of MTTP in the heart, mediated at least partly by the transcription factor CCAAT/enhancer binding protein, and that reduced concentration of MTTP in the myocardium may contribute to IHD upon ischemic damage. PMID- 19546345 TI - Effects of oral estrogen on aortic ROS-generating and -scavenging enzymes and atherosclerosis in apoE-deficient mice. AB - The effect of hormone replacement therapy (HRT) on cardiovascular diseases remains controversial. Studies conducted on postmenopausal women indicate that oral HRT increases risk factors that may counteract the atheroprotective effect of estrogen. However, the effects of estrogen on atherosclerosis have been examined using subcutaneous estrogen in most animal studies, which points to the need for evaluating the effect of oral estrogen. Reactive oxygen species (ROS) have emerged as critical factors in the pathogenesis of atherosclerosis. This study examined the effect of long-term oral estrogen treatment on aortic oxidative stress and atherosclerosis in female apoE(-/-) mice to mimic HRT in humans. Ovariectomized apoE(-/-) mice were given 6 microg/day of oral 17beta estradiol (E(2)) or control vehicle for 12 weeks. Estrogen treatment reduced atherosclerotic lesions by 38% (E(2): 0.20 +/- 0.01 mm(2)/section; control vehicle: 0.32 +/- 0.02 mm(2)/section) and intima by 32% (E(2): 0.44 +/- 0.02 mm(2)/section; control vehicle: 0.65 +/- 0.04 mm(2)/section) in the aortic root. Serum levels of total and low-density lipoprotein cholesterol were significantly decreased after estrogen treatment. Aortic superoxide anion levels and the expression of NAD(P)H oxidase subunit p22(phox) markedly decreased, and two ROS scavenging enzymes, Cu/ZnSOD and MnSOD, were upregulated after estrogen treatment. Estrogen at physiological concentration inhibited tumor necrosis factor-alpha-stimulated NAD(P)H oxidase activity in both cultured smooth muscle cells and peritoneal macrophages. These results showed that long-term oral estrogen treatment reduces ROS levels and atherosclerosis progression in apoE(-/ ) mice. Oral estrogen alters ROS-generating and -scavenging enzyme expression, suggesting that anti-oxidative actions in the vessel wall contribute to atheroprotective effects of estrogen. PMID- 19546346 TI - Gender differences in cardiac ischemic injury and protection--experimental aspects. AB - This review summarizes some available information on gender differences of myocardial injury with particular attention to experimental approach. It has been observed that significant gender differences exist already in normal heart. They involve among others cardiac growth, contractile function, calcium metabolism and function of mitochondria. Differences, characteristic of the normal myocardium, generate the logical presumption of the different reaction of the male and female heart to various pathogenic factors. Most of the experimental studies confirm the clinical observations: increased resistance of the female heart to ischemia/reperfusion injury was shown in dogs, rats, mice and rabbits. Furthermore, gender differences in the ischemic tolerance of the adult myocardium can be influenced by interventions (e.g. hypoxia) imposed during the early phases of ontogenetic development. The already high tolerance of the adult female heart can be increased by adaptation to chronic hypoxia and ischemic preconditioning. It seems that the protective effect depends on age: it was absent in young, highly tolerant heart but it appeared with the decrease of natural resistance during aging. Both experimental and clinical studies have indicated that female gender influences favorably also the remodeling and the adaptive response to myocardial infarction. It follows from the data available that male and female heart differs significantly in many parameters under both physiological and pathological conditions. Detailed molecular and cellular mechanisms of these differences are still unknown; they involve genomic and non-genomic effects of sex steroid hormones, particularly the most frequently studied estrogens. The cardiovascular system is, however, influenced not only by estrogens but also by other sex hormones, e.g. androgens. Moreover, steroid hormone receptors do not act alone but interact with a broad array of co-regulatory proteins to alter transcription. The differences are so important that they deserve serious consideration in clinical practice in search for proper diagnostic and therapeutic procedures. PMID- 19546347 TI - Regulation of glucose uptake in mesangial cells stimulated by high glucose: role of angiotensin II and insulin. AB - Mesangial cells (MCs) play a central role in the pathogenesis of diabetic nephropathy (DN). MC dysfunction arises from excessive glucose uptake through insulin-independent glucose transporter (GLUT1). The role of the insulin dependent transporter (GLUT4) remains unknown. This study evaluated the effect of high glucose on GLUT1, GLUT4, and fibronectin expression levels. Glucose uptake was determined in the absence and presence of insulin. Angiotensin II has been implicated as a mediator of MC abnormalities in DN, and its effects on the GLUTs expression were evaluated in the presence of losartan. MCs were exposed to normal (NG, 10 mM) or high (HG, 30 mM) glucose for 1, 4, 12, 24, and 72 hrs. Glucose uptake was elevated from 1 hr up to 24 hrs of HG, but returned to NG levels after 72 hrs. HG induced an early (1-, 4-, and 12-hrs) rise in GLUT1 expression, returning to NG levels after 72 hrs, whereas GLUT4 was overexpressed at later timepoints (24 and 72 hrs). HG during 4 hrs induced a 40% rise in glucose uptake, which was unaffected by insulin. In contrast, after 72 hrs, glucose uptake was increased by 50%, only under insulin stimulus. Losartan blunted the effects of HG on GLUT1, GLUT4, and fibronectin expression and on glucose uptake. Results suggest that MCs can be highly susceptible to the HG environment since they uptake glucose in both an insulin-independent and insulin-dependent manner. The beneficial effects of angiotensin II inhibition in DN may also involve a decrease in the rate of glucose uptake by MCs. PMID- 19546348 TI - Genetic variations in the hypoxia-inducible factor-1alpha gene and lung cancer. AB - Hypoxia-inducible factor-1 (HIF-1), an important genetic component of angiogenesis, becomes stable as a response to tumor hypoxia and facilitates tumor survival. The polymorphisms of the HIF-1alphagene may cause changes in the activity of this protein, which serves as a transcription factor for many genes in tumorigenesis. In this study, we have investigated the relationship between seven HIF-1alphapolymorphisms [C > T substitution in intron 8 (rs10873142), T418I (rs41508050) in exon 10, P564P (rs41492849), L580L (rs34005929), P582S (rs11549465), A588T (rs11549467) in exon 12 and dinucleotide GT repeats in intron 13 (rs10645014)] among lung cancer patients in the Turkish population. Genomic DNA was isolated from 141 lung cancer cases and 156 controls and subjected to PCR for amplification. Genotyping was carried out with RFLP and DNA sequencing methods. There was no significant difference between the lung cancer cases and controls in terms of the distribution of genotyping frequencies of seven HIF 1alphapolymorphisms (P > 0.05). No significant relationship was found between the C > T substitution in intron 8 and P582S haplotypes and development of lung cancer. In addition, there were no significant associations between the genotypes and clinopathological characteristics of the cases examined. These findings show that polymorphisms in the HIF-1alphagene do not confer susceptibility to lung cancer. PMID- 19546349 TI - Coronavirus diversity, phylogeny and interspecies jumping. AB - The SARS epidemic has boosted interest in research on coronavirus biodiversity and genomics. Before 2003, there were only 10 coronaviruses with complete genomes available. After the SARS epidemic, up to December 2008, there was an addition of 16 coronaviruses with complete genomes sequenced. These include two human coronaviruses (human coronavirus NL63 and human coronavirus HKU1), 10 other mammalian coronaviruses [bat SARS coronavirus, bat coronavirus (bat-CoV) HKU2, bat-CoV HKU4, bat-CoV HKU5, bat-CoV HKU8, bat-CoV HKU9, bat-CoV 512/2005, bat-CoV 1A, equine coronavirus, and beluga whale coronavirus] and four avian coronaviruses (turkey coronavirus, bulbul coronavirus HKU11, thrush coronavirus HKU12, and munia coronavirus HKU13). Two novel subgroups in group 2 coronavirus (groups 2c and 2d) and two novel subgroups in group 3 coronavirus (groups 3b and 3c) have been proposed. The diversity of coronaviruses is a result of the infidelity of RNA-dependent RNA polymerase, high frequency of homologous RNA recombination, and the large genomes of coronaviruses. Among all hosts, the diversity of coronaviruses is most evidenced in bats and birds, which may be a result of their species diversity, ability to fly, environmental pressures, and habits of roosting and flocking. The present evidence supports that bat coronaviruses are the gene pools of group 1 and 2 coronaviruses, whereas bird coronaviruses are the gene pools of group 3 coronaviruses. With the increasing number of coronaviruses, more and more closely related coronaviruses from distantly related animals have been observed, which were results of recent interspecies jumping and may be the cause of disastrous outbreaks of zoonotic diseases. PMID- 19546350 TI - Mitochondrial nitric oxide synthase: a masterpiece of metabolic adaptation, cell growth, transformation, and death. AB - Mitochondria are specialized organelles that control energy metabolism and also activate a multiplicity of pathways that modulate cell proliferation and mitochondrial biogenesis or, conversely, promote cell arrest and programmed cell death by a limited number of oxidative or nitrative reactions. Nitric oxide (NO) regulates oxygen uptake by reversible inhibition of cytochrome oxidase and the production of superoxide anion from the mitochondrial electron transfer chain. In this sense, NO produced by mtNOS will set the oxygen uptake level and contribute to oxidation-reduction reaction (redox)-dependent cell signaling. Modulation of translocation and activation of neuronal nitric oxide synthase (mtNOS activity) under different physiologic or pathologic conditions represents an adaptive response properly modulated to adjust mitochondria to different cell challenges. PMID- 19546351 TI - Relationship between changes in brain MRI and (1)H-MRS, severity of chronic liver damage, and recovery after liver transplantation. AB - Magnetic resonance imaging (MRI) and (1)H magnetic resonance spectroscopy ((1)H MRS) have been used in clinics for diagnosis of chronic liver diseases. This study was designed to investigate the relationship between MRI/MRS outcomes and the severity of liver damage. Of 50 patients examined, the MRI signal intensity in the globus pallidus as determined by pallidus index (PI) increased as the disease severity (scored by Child Pugh ranking) worsened (r = 0.353, P < 0.05). The changes in PI values were also linearly associated with Mn concentrations in whole blood (MnB) (r = 0.814, P < 0.01). MRS analysis of four major brain metabolites (i.e., Cho, mI, Glx, and NAA) revealed that the ratios of Cho/Cr and mI/Cr in cirrhosis and CHE patients were significantly decreased in comparison to controls (P < 0.05), whereas the ratio of Glx/Cr was significantly increased (P < 0.05). The Child Pugh scores significantly correlated with mI/Cr (-0.484, P < 0.01) and Glx (0.369, P < 0.05), as well as MnB (0.368, P < 0.05), but not with other brain metabolites. Three patients who received a liver transplant experienced normalization of brain metabolites within 3 months of post transplantation; the MR imaging of Mn in the globus pallidus completely disappeared 5 months after the surgery. Taken together, this clinical study, which combined MRI/MRS analysis, autopsy exam and liver transplant, clearly demonstrates that liver injury-induced brain Mn accumulation can reversibly alter the homeostasis of brain metabolites Cho, mI and Glx. Our data further suggest that liver transplantation can restore normal brain Mn levels. PMID- 19546352 TI - Dysregulation of T-cell development in adrenal glucocorticoid-deprived rats. AB - A number of different experimental approaches have been used to elucidate the impact of basal levels of adrenal gland-derived glucocorticoids (GCs) on T cell development, and thereby T cell-mediated immune responses. However, the relevance of the adrenal GCs to T cell development is still far from clear. This study was undertaken to explore the relevance of basal levels of GCs to T cell differentiation/maturation. Eight days post-adrenalectomy in adult male rats the thymocyte yield, apoptotic and proliferative rate and the relationship amongst major thymocyte subsets, as defined by TCRalphabeta/CD4/CD8 expression, were examined using flow cytometry. Adrenal GC deprivation decreased thymocyte apoptosis and altered the kinetics of T cell differentiation/maturation. In the adrenalectomized rats there was increased thymic hypercellularity and an over representation of the CD4+CD8+ double positive (DP) TCRalphabeta(low) cells entering selection, as well as increased numbers of their DP TCRalphabeta(-) immediate precursors. These changes were accompanied with under-representation of the postselected DP TCRalphabeta(high) and the most mature CD4-CD8+ and, particularly, CD4+CD8- single positive (SP) TCRalphabeta(high) cells. This data suggests that withdrawal of adrenal GCs produces alterations in the thymocyte selection processes, possibly affecting the diversity of functional T cell repertoire and generation of potentially self-reactive cells as indicated by the reduced proportion and number of CD4-CD8- double negative TCRalphabeta(high) cells. In addition, it indicates that GCs influence the post-selection maturation of thymocytes and plays a regulatory role in controlling the ratio of mature CD4+CD8-/CD4-CD8+ SP TCRalphabeta(high) cells. PMID- 19546353 TI - Increased engraftment of hepatic progenitors after activation of the hepatocyte growth factor signaling pathway by protein transduction. AB - Cell transplantation has become a major focus in biomedical research. However, efficient engraftment in solid tissues remains a challenge. Hepatocyte growth factor (HGF) signaling increases survival, proliferation, migration, and invasion of many cell types through Met, its cell surface receptor. Therefore, activation of this signaling pathway may improve the ability of many cells to be transplanted. We constructed a constitutively activated form of Met (Tpr-Met) fused to the protein transduction domain of HIV-TAT to activate the HGF/Met pathway for a few hours following cell injection. Matrix-assisted refolding was used to renature TAT-Tpr-Met protein, which was efficiently delivered into cells and recapitulated several biological functions of Met in vitro. Furthermore, treatment of hepatic progenitors with this molecule for one hour before transplantation significantly improved engraftment efficiency (31% untreated cells, 58% treated cells). These findings suggest that the transient transfer of Tpr-Met may provide a new approach to increase the proportion of successfully engrafted cells. PMID- 19546354 TI - Sphingolipids as determinants of apoptosis and chemoresistance in the MCF-7 cell model system. AB - An estimated 182,640 women and 1,990 men were diagnosed with breast cancer in 2008, and approximately 40,480 women and 450 men died from the disease. Thus, continued mechanistic studies are needed to understand the causes and develop additional therapeutics for this complicated disease. The MCF-7 cell system is one of the most recognized models for estrogen receptor (ER)-positive breast cancer and has generated approximately 13,000 publications cited in PubMed to date. A number of clues for biological mechanisms related to apoptotic/anti apoptotic pathways and chemoresistance were elucidated and summarized in our previous review. The focus of this review is new knowledge of the central role of sphingolipid signaling in apoptotic mechanisms in estrogen receptor-positive breast cancer. The ultimate goal is to target crucial steps in survival signaling pathways that may ultimately provide additional translational solutions to the successful pharmacologic treatment of breast cancer. PMID- 19546355 TI - TRPC6 up-regulation in Ang II-induced podocyte apoptosis might result from ERK activation and NF-kappaB translocation. AB - Angiotensin II (Ang II) has been recognized as an apoptosis inducer in podocytes, but the mechanism of apoptosis induced by Ang II is unclear. Transient receptor potential cation channel 6 (TRPC6) is a calcium channel located in podocyte membrane. The present study evaluated the alteration of TRPC6 expression and the Ca(2+) influx involved in Ang II-induced podocyte apoptosis. The possible pathways related to TRPC6 in Ang II-induced podocyte apoptosis were also investigated. The apoptosis of mouse podocytes (MPC5) was induced by Ang II. The protein level of TRPC6 was increased markedly in response to Ang II stimulation, and the intracellular Ca(2+) concentration was elevated. By transfection with TRPC6 siRNA, Ang II-induced podocyte apoptosis and the transient Ca(2+) influx were inhibited. Treated with extracellular signal-regulated kinase (ERK) pathway specific inhibitor U0126 or nuclear factor-kappaB (NF-kappaB) pathway specific inhibitor ammonium pyrrolidinedithiocarbamate (PDTC) and Ang II, respectively in podocytes, not only was the TRPC6 up-regulation reduced, but the podocyte apoptosis was also decreased. Moreover, the translocation of NF-kappaB in nucleus resulted from Ang II was reduced by treatment with U0126. In conclusion, the enhancement expression of TRPC6 as well as the increased Ca(2+) influx mediated by TRPC6 channels contributed to the podocyte apoptosis. The activation of ERK pathway and subsequent translocation of NF-kappaB was possibly necessary for the up-regulation TRPC6 induced by Ang II. PMID- 19546356 TI - Propensity of human embryonic stem cell lines during early stage of lineage specification controls their terminal differentiation into mature cell types. AB - Human embryonic stem cells (hESCs) are able to stably maintain their characteristics for an unlimited period; nevertheless, substantial differences among cell lines in gene and protein expression not manifested during the undifferentiated state may appear when cells differentiate. It is widely accepted that developing an efficient protocol to control the differentiation of hESCs will enable us to produce adequate numbers of desired cell types with relative ease for diverse applications ranging from basic research to cell therapy and drug screening. Hence of late, there has been considerable interest in understanding whether and how hESC lines are equivalent or different to each other in their in vitro developmental tendencies. In this study, we compared the developmental competences of two hESC lines (HUES-9 and HUES-7) at molecular, cellular and functional levels, upon spontaneous differentiation without any added inducing agents. Both cell lines generated the three embryonic germ layers, extra-embryonic tissues and primordial germ cells during embryoid body (EB) formation. However HUES-9 showed a stronger propensity towards formation of neuroectodermal lineages, whereas HUES-7 differentiated preferentially into mesoderm and endoderm. Upon further differentiation, HUES-9 generated largely neural cells (neurons, oligodendrocytes, astrocytes and gangliosides) whereas HUES-7 formed mesendodermal derivatives, including cardiomyocytes, skeletal myocytes, endothelial cells, hepatocytes and pancreatic cells. Overall, our findings endorse the hypothesis that independently-derived hESCs biologically differ among themselves, thereby displaying varying differentiation propensity. These subtle differences not only highlight the importance of screening and deriving lines for lineage-specific differentiation but also indicate that individual lines may possess a repertoire of capabilities that is unique. PMID- 19546357 TI - Lanthanum acetate inhibits vascular calcification induced by vitamin D3 plus nicotine in rats. AB - Lanthanum, a rare earth element, has been used to decrease serum phosphorus level in patients with chronic renal disease and hyperphosphatemia. We aimed to observe the effect and mechanism of two doses of lanthanum acetate (375 and 750 mg/kg/day) on vascular calcification induced by vitamin D3 plus nicotine treatment in rats for 4 weeks. As compared with control rats, rats with calcification showed widespread calcified nodules and irregular elastic fibers in calcified aorta on von Kossa calcium staining and increased aortic calcium and phosphorus contents, alkaline phosphatase (ALP) activity and bone-related protein expressions for osteopontin (OPN) and type III sodium dependent phosphate cotransporter Pit-1 (Pit-1). After treatment with either dose of lanthanum acetate, the calcified nodules and degree of irregular elastic fibers decreased in aortas. Lanthanum acetate at 750 mg/kg/day was more effective than 375 mg/kg/day in lessening vascular calcification by significantly reducing plasma phosphorus level, calcium x phosphorus product and ALP activity, by 30.3%, 28.6%, and 68.6%, respectively; reducing aortic phosphorus and calcium contents and ALP activity, by 48%, 53.1%, and 63.5% (all P < 0.01), respectively; reducing aortic mRNA level of OPN and Pit-1, by 55.8% (P < 0.01) and 38.8% (P < 0.05) and protein level of OPN and Pit-1, by 37.2% and 27.2% (both P < 0.01), respectively; and increasing carboxylated matrix Gla-protein (MGP) protein expression by 33.7% (P < 0.05), as compared with rats treated with vitamin D3 and nicotine alone. Lanthanum acetate could effectively inhibit the pathogenesis of vascular calcification. PMID- 19546358 TI - Prevalence and characterization of heterogeneous vancomycin-intermediate Staphylococcus aureus isolates from 14 cities in China. AB - The prevalence of heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) among 1,012 vancomycin-susceptible methicillin (meticillin)-resistant S. aureus isolates collected from 14 cities in China from 2005 to 2007 was 13 to 16%, as determined by a combination of (i) measurement by the modified population analysis profile-area under the curve method (PAP-AUC) and (ii) estimation from the measured sensitivity and specificity of a screening method. Two hundred isolates from blood were chosen as a subset for measurement of the sensitivities and the specificities of several previously described screening methods by using the results of PAP-AUC as the reference. During this testing, one isolate was found to be a vancomycin-intermediate S. aureus (VISA) strain so was not used in the evaluation of the screening tests. Of the other 199 isolates, 26 (13.1%) were hVISA, as assessed by PAP-AUC. A screening cascade of culturing the isolates on brain heart infusion agar containing teicoplanin (5 mg/liter) and then subjecting the positive isolates to a macro-Etest method was applied to the 812 non-blood isolates, yielding 149 positive results. From these results and by adjusting for sensitivity (0.423) and specificity (0.861), the prevalence was estimated to be 15.7%. The precision of that estimate was assessed by reapplying the screening cascade to 120 randomly selected isolates from the 812 non-blood isolates and simultaneously determining their heterogeneous vancomycin-intermediate susceptibility status by PAP-AUC. Because PAP-AUC is impractical for use with large numbers of isolates, the screening-based estimation method is useful as a first approximation of the prevalence of hVISA. Of the 27 VISA or hVISA isolates from blood, 22.2% and 74.1% were staphylococcal chromosome cassette mec types II and III, respectively, while 77.8% and 22.2% were agr type 1 and agr type 2, respectively; the MIC ranges were 0.5 to 4 mg/liter for vancomycin and 0.25 to 1 mg/liter for daptomycin. PMID- 19546359 TI - Pharmacokinetics and buccal mucosal concentrations of a 15 milligram per kilogram of body weight total dose of liposomal amphotericin B administered as a single dose (15 mg/kg), weekly dose (7.5 mg/kg), or daily dose (1 mg/kg) in peripheral stem cell transplant patients. AB - The pharmacokinetics and safety of extended-interval dosing of prophylactic liposomal amphotericin B (L-AMB) in peripheral stem cell transplant recipients were evaluated. The patients received L-AMB daily at 1 mg/kg of body weight or weekly at 7.5 mg/kg or received L-AMB as a single dose (15 mg/kg). The buccal mucosal tissue concentrations of L-AMB were measured. Of the 24 patients enrolled, 5 withdrew after the initial dose due to an infusion-related reaction (n = 2) or significant increases in the serum creatinine (Scr) levels (n = 3). Weekly L-AMB dosing (7.5 mg/kg) produced mean plasma concentrations of >0.300 microg/ml for the first 7 days and >0.220 microg/ml for 7 days after the second dose. A single L-AMB dose (15 mg/kg) produced mean plasma concentrations of >0.491 microg/ml for at least 7 seven days. These concentrations are within the range of the MICs reported in the literature for susceptible strains of Candida and are at the lower limits of the MICs for Aspergillus spp. Extended-interval dosing produced buccal mucosal tissue concentrations well in excess of the MICs reported in the literature for susceptible strains of Candida and Aspergillus spp. Infusion-related reactions occurred in 24% of the patients. Baseline and end of-study Scr, electrolyte (K+, Mg2+, PO4), and serum transaminase levels were similar across the dosage groups. Five (31%) patients met the nephrotoxicity definition prior to completion of the study. Patients in the weekly or single dose groups experienced nephrotoxicity significantly faster than the patients in the daily dosing cohort. A weekly L-AMB dose (7.5 mg/kg) or a single L-AMB dose (15 mg/kg) produced sufficient concentrations in plasma and highly vascular tissue to warrant further studies of the safety, efficacy, and practicality of the weekly prophylactic administration of L-AMB. PMID- 19546360 TI - Antimicrobial susceptibility and mutations involved in clarithromycin resistance in Helicobacter pylori isolates from patients in the western central region of Colombia. AB - Resistance to metronidazole, clarithromycin, and amoxicillin (amoxicilline) was found in 82, 3.8, and 1.9% of 106 Helicobacter pylori isolates, respectively. No tetracycline-resistant isolates were found. In all of the clarithromycin resistant isolates, only one point mutation was present, either A2143G or A2142G. Our results indicate that metronidazole should not be included in the empirical treatment of H. pylori infection in this region. PMID- 19546361 TI - In vitro susceptibilities of Leishmania donovani promastigote and amastigote stages to antileishmanial reference drugs: practical relevance of stage-specific differences. AB - The in vitro susceptibilities of the reference strain Leishmania donovani MHOM/ET/67/L82 to sodium stibogluconate, amphotericin B, miltefosine, and the experimental compound PX-6518 were determined for extracellular log-phase promastigotes, established axenic amastigotes, fresh spleen-derived amastigotes, and intracellular amastigotes in primary mouse peritoneal macrophages. Susceptibility to amphotericin B did not differ across the various axenic models (50% inhibitory concentrations [IC50], 0.6 to 0.7 microM), and amphotericin B showed slightly higher potency against intracellular amastigotes (IC50, 0.1 to 0.4 microM). A similar trend was observed for miltefosine, with comparable efficacies against the extracellular (IC50, 0.4 to 3.8 microM) and intracellular (IC50, 0.9 to 4.3 microM) stages. Sodium stibogluconate, used either as Pentostam or as a crystalline substance, was inactive against all axenic stages (IC50, >64 microg SbV/ml) but showed good efficacy against intracellular amastigotes (IC50, 22 to 28 microg SbV/ml); the crystalline substance was about two to three times more potent (IC50, 9 to 11 microg SbV/ml). The activity profile of PX-6518 was comparable to that of sodium stibogluconate, but at a much higher potency (IC50, 0.1 microg/ml). In conclusion, the differential susceptibility determines which in vitro models are appropriate for either drug screening or resistance monitoring of clinical field isolates. Despite the more complex and labor intensive protocol, the current results support the intracellular amastigote model as the gold standard for in vitro Leishmania drug discovery research and for evaluation of the resistance of field strains, since it also includes host cell-mediated effects. Axenic systems can be recommended only for compounds for which no cellular mechanisms are involved, for example, amphotericin B and miltefosine. PMID- 19546362 TI - Fluorescence-based assay for phenotypic characterization of human cytomegalovirus polymerase mutations regarding drug susceptibility and viral replicative fitness. AB - One essential prerequisite for genotypic drug susceptibility testing of human cytomegalovirus (HCMV) is the phenotypic characterization of mutations identified in the viral protein kinase gene UL97 and the viral DNA polymerase gene UL54 regarding their quantitative impact on drug susceptibility. We developed a new method for phenotypic characterization of UL54 mutations with regard to polymerase activity, viral replication, and drug susceptibility. To determine the most suitable viral indicator gene, enhanced green fluorescence protein was C terminally fused to the HCMV early-late protein UL83 (pp65) or the late proteins UL32 (pp150) and UL99 (pp28), resulting in reporter viruses vTB65g, vTB150g, and vTB28g. vTB65g proved to be superior to the other constructs due to its favorable signal-to-noise ratio and was therefore used to establish the optimum conditions for our assay. The UL54 E756K and D413E mutations were introduced into vTB65g by markerless bacterial artificial chromosome mutagenesis, resulting in virus strains vE756Kg and vD413Eg. The drug susceptibility phenotypes of vE756Kg and vD413Eg were comparable to those previously reported. Furthermore, we found a reduced replicative fitness of vE756Kg by measuring fluorescence intensity as well as by conventional virus growth kinetics. Decreased fluorescence signals of vE756Kg- and vD413Eg-infected cells at late times of infection suggested a reduced polymerase activity, which was confirmed by real-time PCR quantification of the newly synthesized viral DNAs. This new fluorescence-based assay is a highly reproducible method for the phenotypic characterization of mutations potentially influencing drug susceptibility, viral replicative fitness, and polymerase activity of HCMV after marker transfer. PMID- 19546363 TI - Potent activity of a nucleoside reverse transcriptase inhibitor, 4'-ethynyl-2 fluoro-2'-deoxyadenosine, against human immunodeficiency virus type 1 infection in a model using human peripheral blood mononuclear cell-transplanted NOD/SCID Janus kinase 3 knockout mice. AB - 4'-Ethynyl-2-fluoro-2'-deoxyadenosine (EFdA), a recently discovered nucleoside reverse transcriptase inhibitor, exhibits activity against a wide spectrum of wild-type and multidrug-resistant clinical human immunodeficiency virus type 1 (HIV-1) isolates (50% effective concentration, 0.0001 to 0.001 microM). In the present study, we used human peripheral blood mononuclear cell-transplanted, HIV 1-infected NOD/SCID/Janus kinase 3 knockout mice for in vivo evaluation of the anti-HIV activity of EFdA. Administration of EFdA decreased the replication and cytopathic effects of HIV-1 without identifiable adverse effects. In phosphate buffered saline (PBS)-treated mice, the CD4+/CD8+ cell ratio in the spleen was low (median, 0.04; range, 0.02 to 0.49), while that in mice receiving EFdA was increased (median, 0.65; range, 0.57 to 1.43). EFdA treatment significantly suppressed the amount of HIV-1 RNA (median of 9.0 x 10(2) copies/ml [range, 8.1 x 10(2) to 1.1 x 10(3) copies/ml] versus median of 9.9 x 10(4) copies/ml [range, 8.1 x 10(2) to 1.1 x 10(3) copies/ml]; P < 0.001), the p24 level in plasma (2.5 x 10(3) pg/ml [range, 8.2 x 10(2) to 5.6 x 10(3) pg/ml] versus 2.8 x 10(2) pg/ml [range, 8.2 x 10(1) to 6.3 x 10(2) pg/ml]; P < 0.001), and the percentage of p24 expressing cells in the spleen (median of 1.90% [range, 0.33% to 3.68%] versus median of 0.11% [range, 0.00% to 1.00%]; P = 0.003) in comparison with PBS treated mice. These data suggest that EFdA is a promising candidate for a new age of HIV-1 chemotherapy and should be developed further as a potential therapy for individuals with multidrug-resistant HIV-1 variants. PMID- 19546364 TI - In vitro and in vivo properties of dihydrophthalazine antifolates, a novel family of antibacterial drugs. AB - Racemic 2,4-diaminopyrimidine dihydrophthalazine derivatives BAL0030543, BAL0030544, and BAL0030545 exhibited low in vitro MICs toward small, selected panels of Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae, Moraxella catarrhalis, and Mycobacterium avium, though the compounds were less active against Haemophilus influenzae. The constellation of dihydrofolate reductases (DHFRs) present in 20 enterococci and 40 staphylococci was analyzed and correlated with the antibacterial activities of the dihydrophthalazines and trimethoprim. DHFRs encoded by dfrB, dfrA (S1 isozyme), dfrE, and folA were susceptible to the dihydrophthalazines, whereas DHFRs encoded by dfrG (S3 isozyme) and dfrF were not. Studies with the separated enantiomers of BAL0030543, BAL0030544, and BAL0030545 revealed preferential inhibition of susceptible DHFRs by the (R)-enantiomers. BAL0030543, BAL0030544, and BAL0030545 were well tolerated by mice during 5- and 10-day oral toxicity studies at doses of up to 400 mg/kg of body weight. Using a nonoptimized formulation, the dihydrophthalazines displayed acceptable oral bioavailabilities in mice, and efficacy studies with a septicemia model of mice infected with trimethoprim resistant, methicillin-resistant Staphylococcus aureus gave 50% effective dose values in the range of 1.6 to 6.25 mg/kg. PMID- 19546365 TI - Emergence of extended-spectrum beta-lactamase PER-1 in Proteus vulgaris and Providencia stuartii isolates from Algiers, Algeria. PMID- 19546366 TI - Influence of recombination on development of mutational resistance to linezolid in Enterococcus faecalis JH2-2. AB - We compared the propensities of Enterococcus faecalis JH2-2 and of the recombination-deficient JH2-2 recA strain to develop mutational resistance to linezolid. In both organisms, a mutation in a single rrl copy conferred resistance to linezolid. Delay in acquisition of the mutation by other rrl copies in JH2-2 recA showed that gene conversion contributed to the acquisition of resistance. PMID- 19546367 TI - Effect of Candida glabrata FKS1 and FKS2 mutations on echinocandin sensitivity and kinetics of 1,3-beta-D-glucan synthase: implication for the existing susceptibility breakpoint. AB - Thirteen Candida glabrata strains harboring a range of mutations in hot spot regions of FKS1 and FKS2 were studied. The mutations were linked to an echinocandin reduced susceptibility phenotype. Sequence alignments showed that 11 out of the 13 mutants harbored a mutation in FKS1 or FKS2 not previously implicated in echinocandin reduced susceptibility in C. glabrata. A detailed kinetic characterization demonstrated that amino acid substitutions in Fks1p and Fks2p reduced drug sensitivity in mutant 1,3-beta-D-glucan synthase by 2 to 3 log orders relative to that in wild-type enzyme. These mutations were also found to reduce the catalytic efficiency of the enzyme (Vmax) and to influence the relative expression of FKS genes. In view of the association of FKS mutations and reduced susceptibility of 1,3-beta-D-glucan synthase, an evaluation of the new CLSI echinocandin susceptibility breakpoint was conducted. Only 3 of 13 resistant fks mutants (23%) were considered anidulafungin or micafungin nonsusceptible (MIC > 2 microg/ml) by this criterion. In contrast, most fks mutants (92%) exceeded a MIC of >2 microg/ml with caspofungin. However, when MIC determinations were performed in the presence of 50% serum, all C. glabrata fks mutants showed MICs of > or = 2 microg/ml for the three echinocandin drugs. As has been observed with Candida albicans, the kinetic inhibition parameter 50% inhibitory concentration may be a better predictor of FKS-mediated resistance. Finally, the close association between FKS1/FKS2 hot spot mutations provides a basis for understanding echinocandin resistance in C. glabrata. PMID- 19546368 TI - Biofilm formation and effect of caspofungin on biofilm structure of Candida species bloodstream isolates. AB - Candida biofilms are microbial communities, embedded in a polymeric matrix, growing attached to a surface, and are highly recalcitrant to antimicrobial therapy. These biofilms exhibit enhanced resistance against most antifungal agents except echinocandins and lipid formulations of amphotericin B. In this study, biofilm formation by different Candida species, particularly Candida albicans, C. tropicalis, and C. parapsilosis, was evaluated, and the effect of caspofungin (CAS) was assessed using a clinically relevant in vitro model system. CAS displayed in vitro activity against C. albicans and C. tropicalis cells within biofilms. Biofilm formation was evaluated after 48 h of antifungal drug exposure, and the effects of CAS on preformed Candida species biofilms were visualized using scanning electron microscopy (SEM). Several species-specific differences in the cellular morphologies associated with biofilms were observed. Our results confirmed the presence of paradoxical growth (PG) in C. albicans and C. tropicalis biofilms in the presence of high CAS concentrations. These findings were also confirmed by SEM analysis and were associated with the metabolic activity obtained by biofilm susceptibility testing. Importantly, these results suggest that the presence of atypical, enlarged, conical cells could be associated with PG and with tolerant cells in Candida species biofilm populations. The clinical implications of these findings are still unknown. PMID- 19546369 TI - A novel tryptophanyl-tRNA synthetase gene confers high-level resistance to indolmycin. AB - Indolmycin, a potential antibacterial drug, competitively inhibits bacterial tryptophanyl-tRNA synthetases. An effort to identify indolmycin resistance genes led to the discovery of a gene encoding an indolmycin-resistant isoform of tryptophanyl-tRNA synthetase. Overexpression of this gene in an indolmycin sensitive strain increased the indolmycin MIC 60-fold. Its transcription and distribution in various bacterial genera were assessed. The level of resistance conferred by this gene was compared to that of a known indolmycin resistance gene and to those of genes with resistance-conferring point mutations. PMID- 19546370 TI - Mosaic penicillin-binding protein 2 in Neisseria gonorrhoeae isolates collected in 2008 in San Francisco, California. AB - Using a real-time PCR assay specific for a mosaic penA allele that has been associated with oral cephalosporin resistance in Asia, 54 available Neisseria gonorrhoeae isolates collected in San Francisco, CA, from January to October 2008 were analyzed. Five isolates tested positive for the mosaic penA gene by real time PCR. DNA sequencing revealed two mosaic penA alleles (SF-A and SF-B). Isolates with SF-A and SF-B alleles possessed elevated MICs for the oral cephalosporins cefpodoxime and cefixime. PMID- 19546371 TI - Evaluation of target specificity of antibacterial agents using Staphylococcus aureus ddlA mutants and D-cycloserine in a silkworm infection model. AB - The availability of a silkworm larva infection model to evaluate the therapeutic effectiveness of antibiotics was examined. The 50% effective doses (ED50) of D cycloserine against the Staphylococcus aureus ddlA mutant-mediated killing of larvae were remarkably lower than those against the parental strain-mediated killing of larvae. Changes in MICs and ED50 of other antibiotics were negligible, suggesting that these alterations are d-cycloserine selective. Therefore, this model is useful for selecting desired compounds based on their therapeutic effectiveness during antibiotic development. PMID- 19546372 TI - Novel expression vector for secretion of cecropin AD in Bacillus subtilis with enhanced antimicrobial activity. AB - Cecropin AD, a chimeric antimicrobial peptide obtained from cecropins, is effective at killing specific microorganisms. However, a highly efficient expression system is still needed to allow for commercial application of cecropin AD. For the exogenous expression of cecropin AD, we fused the cecropin AD gene with a small ubiquitin-like modifier (SUMO) gene and a signal peptide of SacB, while a Bacillus subtilis expression system was constructed based on Bacillus subtilis cells genetically modified by the introduction of an operon including an isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible Spac promoter, a signal peptide of amyQ, and the SUMO protease gene. The recombinant cecropin AD was expressed, and 30.6 mg of pure recombinant protein was purified from 1 liter of culture supernatant. The purified cecropin AD displayed antimicrobial activity against some pathogens, such as Staphylococcus aureus and Escherichia coli, and was especially effective toward Staphylococcus aureus, with MICs of <0.05 microM (0.2 microg/ml). Stability analysis results showed that the activity of cecropin AD was not influenced by temperatures as high as 55 degrees C for 20 min; however, temperatures above 85 degrees C (for 20 min) decreased the antimicrobial activity of cecropin AD. Varying the pH from 4.0 to 9.0 did not appear to affect the activity of cecropin AD, but some loss of potency was observed at pH values lower than pH 4.0. Under the challenge of several proteases (proteinase K, trypsin, and pepsin), cecropin AD maintained functional activity. The results indicated that the recombinant product expressed by the designed Bacillus subtilis expression system was a potent antimicrobial agent and could be applied to control infectious diseases of farm animals or even humans. PMID- 19546373 TI - (Pro)renin receptor and vacuolar H+-ATPase. PMID- 19546374 TI - Impact of shear rate modulation on vascular function in humans. AB - Shear stress is an important stimulus to arterial adaptation in response to exercise and training in humans. We recently observed significant reverse arterial flow and shear during exercise and different antegrade/retrograde patterns of shear and flow in response to different types of exercise. The purpose of this study was to simultaneously examine flow-mediated dilation, a largely NO-mediated vasodilator response, in both brachial arteries of healthy young men before and after 30-minute interventions consisting of bilateral forearm heating, recumbent leg cycling, and bilateral handgrip exercise. During each intervention, a cuff inflated to 60 mm Hg was placed on 1 arm to unilaterally manipulate the shear rate stimulus. In the noncuffed arm, antegrade flow and shear increased similarly in response to each intervention (ANOVA; P<0.001, no interaction between interventions; P=0.71). Baseline flow-mediated dilation (4.6%, 6.9%, and 6.7%) increased similarly in response to heating, handgrip, and cycling (8.1%, 10.4%, and 8.9%, ANOVA; P<0.001, no interaction; P=0.89). In contrast, cuffed arm antegrade shear rate was lower than in the noncuffed arm for all of the conditions (P<0.05), and the increase in flow mediated dilation was abolished in this arm (4.7%, 6.7%, and 6.1%; 2-way ANOVA: all conditions interacted P<0.05). These results suggest that differences in the magnitude of antegrade shear rate transduce differences in endothelial vasodilator function in humans, a finding that may have relevance for the impact of different exercise interventions on vascular adaptation in humans. PMID- 19546375 TI - Protective importance of the myogenic response in the renal circulation. PMID- 19546376 TI - Large and small artery cross-talk and recent morbidity-mortality trials in hypertension. PMID- 19546377 TI - Amplification of the pressure pulse in the upper limb in healthy, middle-aged men and women. AB - Central-to-peripheral amplification of the pressure pulse leads to discrepancies between central and brachial blood pressures. This amplification depends on an individual's hemodynamic and (patho)physiological characteristics. The aim of this study was to assess the magnitude and correlates of central-to-peripheral amplification in the upper limb in a healthy, middle-aged population (the Asklepios Study). Carotid, brachial, and radial pressure waveforms were acquired noninvasively using applanation tonometry in 1873 subjects (895 women) aged 35 to 55 years. Carotid, brachial, and radial pulse pressures were calculated, as well as the absolute and relative (with carotid pulse pressure as reference) amplifications. With subjects classified per semidecade of age, carotid-to-radial amplification varied from approximately 25% in the youngest men to 8% in the oldest women. Amplification was higher in men (20+/-14%) than in women (13+/-12%; P<0.001) and decreased with age (P<0.001) in both. Amplification over the brachial-to-radial path contributed substantially to the total amplification. In univariate analysis, the strongest correlation was found with the carotid augmentation index (-0.51 in women; -0.47 in men; both P<0.001). In a multiple linear regression model with carotid-to-radial amplification as the dependent variable, carotid augmentation index, total arterial compliance, and heart rate were identified as the 3 major determinants of upper limb pressure amplification (R2=0.36). We conclude that, in healthy middle-aged subjects, the central-to radial amplification of the pressure pulse is substantial. Amplification is higher in men than in women, decreases with age, and is primarily associated with the carotid augmentation index. PMID- 19546378 TI - Novel role of fumarate metabolism in dahl-salt sensitive hypertension. AB - In a previous proteomic study, we found dramatic differences in fumarase in the kidney between Dahl salt-sensitive rats and salt-insensitive consomic SS-13(BN) rats. Fumarase catalyzes the conversion between fumarate and l-malate in the tricarboxylic acid cycle. Little is known about the pathophysiological significance of fumarate metabolism in cardiovascular and renal functions, including salt-induced hypertension. The fumarase gene is located on the chromosome substituted in the SS-13(BN) rat. Sequencing of fumarase cDNA indicated the presence of lysine at amino acid position 481 in Dahl salt sensitive rats and glutamic acid in Brown Norway and SS-13(BN) rats. Total fumarase activity was significantly lower in the kidneys of Dahl salt-sensitive rats compared with SS-13(BN) rats, despite an apparent compensatory increase in fumarase abundance in Dahl salt-sensitive rats. Intravenous infusion of a fumarate precursor in SS-13(BN) rats resulted in a fumarate excess in the renal medulla comparable to that seen in Dahl salt-sensitive rats. The infusion significantly exacerbated salt-induced hypertension in SS-13(BN) rats (140+/-3 vs125+/-2 mm Hg in vehicle control at day 5 on a 4% NaCl diet; P<0.05). In addition, the fumarate infusion increased renal medullary tissue levels of H2O2. Treatment of cultured human renal epithelial cells with the fumarate precursor also increased cellular levels of H2O2. These data suggest a novel role for fumarate metabolism in salt-induced hypertension and renal medullary oxidative stress. PMID- 19546380 TI - The (Pro)renin receptor: site-specific and functional linkage to the vacuolar H+ ATPase in the kidney. AB - The (pro)renin receptor ([P]RR) is a transmembrane protein that binds both renin and prorenin with high affinity, increasing the catalytic cleavage of angiotensinogen and signaling intracellularly through mitogen-activated protein kinase activation. Although initially reported as having no homology with any known membrane protein, other studies have suggested that the (P)RR is an accessory protein, named ATP6ap2, that associates with the vacuolar H(+)-ATPase, a key mediator of final urinary acidification. Using in situ hybridization, immunohistochemistry, and electron microscopy, together with serial sections stained with nephron segment-specific markers, we found that (P)RR mRNA and protein were predominantly expressed in collecting ducts and in the distal nephron. Within collecting ducts, the (P)RR was most abundant in microvilli at the apical surface of A-type intercalated cells. Dual-staining immunofluorescence demonstrated colocalization of the (P)RR with the B1/2 subunit of the vacuolar H(+)-ATPase, the ion exchanger that secretes H(+) ions into the urinary space and that associates with an accessory subunit homologous to the (P)RR. In collecting duct/distal tubule lineage Madin-Darby canine kidney cells, extracellular signal regulated kinase 1/2 phosphorylation, induced by either renin or prorenin, was attenuated by the selective vacuolar H(+)-ATPase inhibitor bafilomycin. The predominant expression of the (P)RR at the apex of acid-secreting cells in the collecting duct, along with its colocalization and homology with an accessory protein of the vacuolar H(+)-ATPase, suggests that the (P)RR may function primarily in distal nephron H(+) transport, recently noted to be, at least in part, an angiotensin II-dependent phenomenon. PMID- 19546379 TI - Failures in mitochondrial tRNAMet and tRNAGln metabolism caused by the novel 4401A>G mutation are involved in essential hypertension in a Han Chinese Family. AB - We report here on the clinical, genetic, and molecular characterization of 1 Han Chinese family with maternally transmitted hypertension. Three of 7 matrilineal relatives in this 4-generation family exhibited the variable degree of essential hypertension at the age at onset, ranging from 35 to 60 years old. Sequence analysis of the complete mitochondrial DNA in this pedigree identified the novel homoplasmic 4401A>G mutation localizing at the spacer immediately to the 5' end of tRNA(Met) and tRNA(Gln) genes and 39 other variants belonging to the Asian haplogroup C. The 4401A>G mutation was absent in 242 Han Chinese controls. Approximately 30% reductions in the steady-state levels of tRNA(Met) and tRNA(Gln) were observed in 2 lymphoblastoid cell lines carrying the 4401A>G mutation compared with 2 control cell lines lacking this mutation. Failures in mitochondrial metabolism are apparently a primary contributor to the reduced rate of mitochondrial translation and reductions in the rate of overall respiratory capacity, malate/glutamate-promoted respiration, succinate/glycerol-3-phosphate promoted respiration, or N,N,N',N'-tetramethyl-p-phenylenediamine/ascorbate promoted respiration in lymphoblastoid cell lines carrying the 4401A>G mutation. The homoplasmic form, mild biochemical defect, late onset, and incomplete penetrance of hypertension in this family suggest that the 4401A>G mutation itself is insufficient to produce a clinical phenotype. Thus, the other modifier factors, eg, nuclear modifier genes and environmental and personal factors, may also contribute to the development of hypertension in these subjects carrying this mutation. These data suggest that mitochondrial dysfunctions, caused by the 4401A>G mutation, are involved in the development of hypertension in this Chinese pedigree. PMID- 19546382 TI - Kawasaki disease: a childhood disease with important consequences into adulthood. PMID- 19546381 TI - Arginase II knockout mouse displays a hypertensive phenotype despite a decreased vasoconstrictory profile. AB - Arginase upregulation is associated with aging and cardiovascular diseases. In this study we report on the cardiovascular phenotype of the arginase II knockout (KO) mouse. We demonstrate that vascular sensitivity and reactivity altered over time in these animals such that no influence on responses to vasoconstrictor activity was observed in 7-week-old KO mice, but dampened responses to norepinephrine and phenylephrine were observed by 10 and 15 weeks with Rho kinase influencing these effects in the 15-week-old animals. Despite these dampened vasoconstrictory responses, KO mice demonstrated increased mean arterial pressure from 8 weeks old. This hypertensive phenotype was associated with an increase in left ventricular weight, left ventricular systolic pressure, and diminished diastolic function. KO mice also show enhanced plasma norepinephrine turnover, suggesting an increased sympathetic outflow. In conclusion, our data suggest that global loss of arginase II activity results in hypertension. We suggest that this strain of mouse warrants further investigation as a potentially novel model of hypertension. PMID- 19546383 TI - Improving lipid goal attainment: is it enough? PMID- 19546384 TI - Twenty-five-year outcome of pediatric coronary artery bypass surgery for Kawasaki disease. AB - BACKGROUND: The long-term outcome of pediatric coronary artery bypass for patients with severe inflammatory coronary sequelae secondary to Kawasaki disease is unknown. METHODS AND RESULTS: One hundred fourteen children and adolescents ranging in age from 1 to 19 (median, 10) years at operation were followed up for as long as 25 years with a median of 19 years. The number of distal anastomoses was 1.7+/-0.8 per patient, and the internal thoracic artery was used in all but 3, most frequently for left anterior descending artery lesions. Saphenous vein grafts were used in 24 patients, mostly for non-left anterior descending artery lesions. Patients underwent multiple angiograms to evaluate their coronary and graft status. There was no operative or hospital mortality. Both 20- and 25-year survival rates were 95% (95% confidence interval [CI], 88 to 98). Five deaths occurred, all cardiac in origin. Cardiac event-free rates at 20 and 25 years were 67% and 60% (95% CI, 46 to 72), respectively. Percutaneous coronary intervention and reoperation were the most common events. Overall, the 20-year graft patency rate was 87% (95% CI, 78 to 93) for internal thoracic artery grafts (n=154) and 44% (95% CI, 26 to 61) for saphenous vein grafts (n=30) (P<0.001), and the rate for non-left anterior descending artery lesions was also significantly better for arterial grafts (87% [95% CI, 73 to 94]; n=59) than for saphenous vein grafts (42% [95% CI, 23 to 60]; n=27) (P=0.002). Eighty-eight patients (77%) remain on medications, but all 109 survivors are presently symptom free in their daily activities. CONCLUSIONS: Although the 25-year survival was excellent after pediatric coronary bypass for Kawasaki disease, the event-free rate declined progressively. This reality mandated continued follow-up. Reinterventions successfully managed most cardiac events. An internal thoracic artery graft was the most favorable for children. PMID- 19546385 TI - Visually-guided balloon catheter ablation of atrial fibrillation: experimental feasibility and first-in-human multicenter clinical outcome. AB - BACKGROUND: Electric isolation of the pulmonary veins (PVs) can successfully treat patients with paroxysmal atrial fibrillation. However, it remains technically challenging to identify the left atrial-PV junction and sequentially position the ablation catheter in a point-by-point contiguous fashion to isolate the PVs. In this study, a novel endoscopic ablation system was used to directly visualize and ablate tissue at the left atrial-PV junction with laser energy. METHODS AND RESULTS: This study consisted of 2 phases: a short-term (n=9) and long-term (n=11) canine experimental validation phase and a multicenter clinical feasibility phase (n=30 paroxysmal atrial fibrillation patients). After transseptal puncture, the balloon-based endoscopic ablation system was advanced to each PV ostium, and arcs of laser energy (90 degrees to 360 degrees ) were projected onto the target left atrial-PV junction. Electric PV isolation was defined with a circular multielectrode catheter. In the short-term preclinical experimental phase, 15 of 17 targeted PVs (88%) were successfully isolated. Pathological examination revealed well-demarcated circumferential lesions with minimal endothelial disruption. In the long-term experiments, 9 of 10 targeted veins (90%) remained persistently isolated (at 4 to 8 weeks). In the clinical phase, 105 of 116 PVs (91%) were successfully isolated. After a single procedure, the 12-month drug-free rate of freedom from atrial fibrillation was 60% (18 of 30 patients). There were no significant PV stenoses, but adverse events included 1 episode of cardiac tamponade, 1 stroke without residual defect, and 1 asymptomatic phrenic nerve palsy. CONCLUSIONS: This study establishes the feasibility of a novel paradigm for AF ablation: direct visualization to guide catheter ablation of the left atrial-PV junction. PMID- 19546386 TI - Lipid treatment assessment project 2: a multinational survey to evaluate the proportion of patients achieving low-density lipoprotein cholesterol goals. AB - BACKGROUND: Information about physicians' adherence to cholesterol management guidelines remains scant. The present survey updates our knowledge of lipid management worldwide. METHODS AND RESULTS: Lipid levels were determined at enrollment in dyslipidemic adult patients on stable lipid-lowering therapy in 9 countries. The primary end point was the success rate, defined as the proportion of patients achieving appropriate low-density lipoprotein cholesterol (LDL-C) goals for their given risk. The mean age of the 9955 evaluable patients was 62+/ 12 years; 54% were male. Coronary disease and diabetes mellitus had been diagnosed in 30% and 31%, respectively, and 14% were current smokers. Current treatment consisted of a statin in 75%. The proportion of patients achieving LDL C goals according to relevant national guidelines ranged from 47% to 84% across countries. In low-, moderate-, and high-risk groups, mean LDL-C was 119, 109, and 91 mg/dL and mean high-density lipoprotein cholesterol was 62, 49, and 50 mg/dL, respectively. The success rate for LDL-C goal achievement was 86% in low-, 74% in moderate-, and 67% in high-risk patients (73% overall). However, among coronary heart disease patients with > or =2 risk factors, only 30% attained the optional LDL-C goal of <70 mg/dL. In the entire cohort, high-density lipoprotein cholesterol was <40 mg/dL in 19%, 40 to 60 mg/dL in 55%, and >60 mg/dL in 26% of patients. CONCLUSIONS: Although there is room for improvement, particularly in very-high-risk patients, these results indicate that lipid-lowering therapy is being applied much more successfully than it was a decade ago. PMID- 19546387 TI - Safety and efficacy of recombinant activated factor VII: a randomized placebo controlled trial in the setting of bleeding after cardiac surgery. AB - BACKGROUND: Blood loss is a common complication of cardiac surgery. Evidence suggests that recombinant activated factor VII (rFVIIa) can decrease intractable bleeding in patients after cardiac surgery. Our objective was to investigate the safety and possible benefits of rFVIIa in patients who bleed after cardiac surgery. METHODS AND RESULTS: In this phase II dose-escalation study, patients who had undergone cardiac surgery and were bleeding were randomized to receive placebo (n=68), 40 microg/kg rFVIIa (n=35), or 80 microg/kg rFVIIa (n=69). The primary end points were the number of patients suffering critical serious adverse events. Secondary end points included rates of reoperation, amount of blood loss, and transfusion of allogeneic blood. There were more critical serious adverse events in the rFVIIa groups. These differences did not reach statistical significance (placebo, 7%; 40 microg/kg, 14%; P=0.25; 80 microg/kg, 12%; P=0.43). After randomization, significantly fewer patients in the rFVIIa group underwent a reoperation as a result of bleeding (P=0.03) or required allogeneic transfusions (P=0.01). CONCLUSIONS: On the basis of this preliminary evidence, rFVIIa may be beneficial for treating bleeding after cardiac surgery, but caution should be applied and further clinical trials are required because there is an increase in the number of critical serious adverse events, including stroke, in those patients randomized to receive rFVIIa. PMID- 19546388 TI - Opposite effects of training in rats with stable and progressive pulmonary hypertension. AB - BACKGROUND: Exercise training in pulmonary arterial hypertension (PH) is a promising adjunct to medical treatment. However, it is still unclear whether training is beneficial for all PH patients. We hypothesized that right ventricular adaptation plays a pivotal role in the response to training. METHODS AND RESULTS: Two different dosages of monocrotaline were used in rats to model stable PH with preserved cardiac output and progressive PH developing right heart failure. Two weeks after injection, PH was confirmed by echocardiography, and treadmill training was initiated. Rats were trained for 4 weeks unless manifest right heart failure developed earlier. At the end of the study protocol, all rats were functionally assessed by endurance testing, echocardiography, and invasive pressure measurements. Lungs and hearts were further analyzed in quantitative histomorphologic analyses. In stable PH, exercise training was well tolerated and markedly increased exercise endurance (from 25+/-3.9 to 62+/-3.9 minutes; P<0.001). Moreover, capillary density increased significantly (from 1.21+/-0.12 to 1.51+/-0.07 capillaries per cardiomyocyte; P<0.05). However, in progressive PH, exercise training worsened survival (hazard ratio, 2.7; 95% confidence interval, 1.1 to 14.2) and increased pulmonary vascular remodeling. In addition, training induced widespread leukocyte infiltration into the right ventricle (from 135+/-14 to 276+/-18 leukocytes per 1 mm(2); P<0.001). CONCLUSIONS: In our rat model, exercise training was found to be beneficial in stable PH but detrimental in progressive PH. Future studies are necessary to address the clinical implications of our findings. PMID- 19546389 TI - Urinary N-terminal prohormone brain natriuretic peptide excretion in patients with chronic heart failure. AB - BACKGROUND: Urinary excretion is currently regarded as the main mechanism of elimination of N-terminal prohormone brain natriuretic peptide (NT-proBNP). The clinical implications and the value of measurement of urinary NT-proBNP in patients with heart failure are largely unknown. METHODS AND RESULTS: We studied 94 patients (age, 58+/-11 years; 79% men) with chronic heart failure (CHF) and 20 age- and sex-matched healthy control subjects. Glomerular filtration rate and effective renal plasma flow were measured as clearance of (125)I-iothalamate and (131)I-hippuran, respectively. NT-proBNP levels were determined in both plasma and 24-hour urine collections. Mean left ventricular ejection fraction of CHF patients was 0.28+/-0.09. Plasma NT-proBNP levels were higher in CHF patients compared with control subjects (median, 547 versus 41 pg/mL; P<0.001). Urinary NT proBNP excretion, however, was substantially lower in CHF patients (median, 0.13 versus 2.3 mL/min; P<0.001). Urinary NT-proBNP excretion was independent of estimated glomerular filtration rate. In both CHF patients and control subjects, there was a strong and inverse relation between plasma NT-proBNP concentrations and urinary NT-proBNP excretion (r=-0.72 and r=-0.65 respectively; both P<0.001). Decreased renal plasma flow in CHF was significantly associated with a lower excretion of NT-proBNP (P=0.026). CONCLUSIONS: Urinary NT-proBNP excretion is lower in patients with CHF compared with control subjects and is inversely related to plasma NT-proBNP. Urinary NT-proBNP is associated with renal plasma flow but not with estimated glomerular filtration rate. Elevated levels of plasma NT-proBNP in patients with CHF might be explained not only by myocardial stress but also by a marked decrease in urinary excretion. PMID- 19546391 TI - Risk score for predicting outcome in patients with asymptomatic aortic stenosis. AB - BACKGROUND: The management of patients with asymptomatic severe aortic stenosis remains controversial. We sought to develop a continuous risk score for predicting the midterm development of symptoms or adverse events in this setting. METHODS AND RESULTS: We prospectively followed 107 patients with asymptomatic aortic stenosis (aged 72 years [63 to 77]; 35 women; aortic-jet velocity, 4.1 m/s [3.5 to 4.4]) at a single center in France. Predefined end points for assessing outcome were the occurrence within 24 months of death or aortic valve replacement necessitated by symptoms or by a positive exercise test. Variables independently associated with outcome were used to build a score that was validated in an independent cohort of 107 patients from Belgium. Independent predictors of outcome were female sex, peak aortic-jet velocity, and B-type natriuretic peptide at baseline. Accordingly, the score could be calculated as follows: Score=[peak velocity (m/s) x 2]+(natural logarithm of B-type natriuretic peptide x 1.5)+1.5 (if female sex). Event-free survival after 20 months was 80% for patients within the first score quartile compared with only 7% for the fourth quartile. Areas under the receiver operating characteristic curve for the score were 0.90 and 0.89 in the development and validation cohorts, respectively. CONCLUSIONS: If further validation is achieved, this score may be useful to predict outcome in individual patients with asymptomatic aortic stenosis to select those who might benefit from early surgery. PMID- 19546390 TI - Inhibition of prolyl hydroxylase domain proteins promotes therapeutic revascularization. AB - BACKGROUND: The hypoxia-inducible transcription factor (HIF) subunits are destabilized via the O(2)-dependent prolyl hydroxylase domain proteins (PHD1, PHD2, and PHD3). We investigated whether inhibition of PHDs via upregulating HIF might promote postischemic neovascularization. METHODS AND RESULTS: Mice with right femoral artery ligation were treated, by in vivo electrotransfer, with plasmids encoding for an irrelevant short hairpin RNA (shRNA) (shCON [control]) or specific shRNAs directed against HIF-1alpha (shHIF-1alpha), PHD1 (shPHD1), PHD2 (shPHD2), and PHD3 (shPHD3). The silencing of PHDs induced a specific and transient downregulation of their respective mRNA and protein levels at day 2 after ischemia and, as expected, upregulated HIF-1alpha. As a consequence, 2 key hypoxia-inducible proangiogenic actors, vascular endothelial growth factor-A and endothelial nitric oxide synthase, were upregulated at the mRNA and protein levels. In addition, monocyte chemotactic protein-1 mRNA levels and infiltration of Mac-3-positive macrophages were enhanced in ischemic leg of mice treated with shPHD2 and shPHD3. Furthermore, activation of HIF-1alpha-related pathways was associated with changes in postischemic neovascularization. At day 14, silencing of PHD2 and PHD3 increased vessel density by 2.2- and 2.6-fold, capillary density by 1.8- and 2.1-fold, and foot perfusion by 1.2- and 1.4-fold, respectively, compared with shCON (P<0.001). shPHD1 displayed a lower proangiogenic effect. Of interest, coadministration of shHIF-1alpha with shPHD3 abrogated shPHD3-related effects, suggesting that activation of endogenous HIF-1-dependent pathways mediated the proangiogenic effects of PHD silencing. CONCLUSIONS: We demonstrated that a direct inhibition of PHDs, and more particularly PHD3, promoted therapeutic revascularization. Furthermore, we showed that activation of the HIF 1 signaling pathway is required to promote this revascularization. PMID- 19546393 TI - The Lewis lead: making recognition of P waves easy during wide QRS complex tachycardia. PMID- 19546394 TI - Letter by cattaneo regarding article, "incomplete inhibition of thromboxane biosynthesis by acetylsalicylic Acid: determinants and effect on cardiovascular risk". PMID- 19546396 TI - Insights into the role of infection in atherogenesis and in plaque rupture. PMID- 19546397 TI - Forty-year-old intrathoracic gossypiboma after cardiac valve surgery. PMID- 19546399 TI - Stopping trials early for positive results: the need to know how much. PMID- 19546401 TI - Natural History of CNS Relapse in Aggressive Non-Hodgkin's Lymphoma: What Have We Learned? PMID- 19546403 TI - Secondary causes of low bone mass in patients with breast cancer: a need for greater vigilance. AB - PURPOSE: An observational study to assess the prevalence of secondary causes of low bone density in patients with breast cancer. PATIENTS AND METHODS: Female patients within 5 years of breast cancer diagnosis and age older than 50 years at diagnosis were recruited. Consenting patients completed a questionnaire and had blood taken for serum calcium, vitamin D, parathyroid hormone (PTH), and thyroid function testing. Bone mineral density (BMD) was assessed. Abnormalities were additionally investigated and treated. RESULTS: Two hundred patients were recruited. The median age at diagnosis was 62 years. One hundred sixty-nine patients had hormone receptor (HR) -positive cancer. Vitamin D and thyroid function were assessed in 200 patients; PTH was assessed in 197 patients; and BMD was assessed in 187 patients. Eighty-seven patients (46.5%) had osteopenia, and 24 patients (12.8%) had osteoporosis. Vitamin D levels were insufficient (ie, 50 to 75 nmol/L) in 74 patients (37%) and were deficient (ie, < 50 nmol/L) in 54 patients (27%). Only 24 of 65 patients taking vitamin D supplements were replete (ie, > 75 nmol/L). Thirty-nine (21%) of 197 patients had PTH concentrations greater than the normal range. Six had primary hyperparathyroidism (PHPT), and two more had recent surgery for PHPT. Twenty-seven had secondary hyperparathyroidism (HPT) from vitamin D deficiency and six had normocalcemic HPT. Of 90 patients with low BMD and HR-positive cancer, 8% (seven of 90 patients) had new or recent PHPT, and 63% (57 of 90 patients) had insufficient or deficient vitamin D. More patients with HR-positive cancer than with HR-negative cancer had elevated PTH (38 of 167 v three of 30 patients, respectively; P = .10). CONCLUSION: Secondary causes of low BMD are common in postmenopausal women. Identification and management of secondary causes should be included in bone health management algorithms. PMID- 19546402 TI - Minimal disseminated disease in childhood T-cell lymphoblastic lymphoma: a report from the children's oncology group. AB - PURPOSE Disease dissemination to the bone marrow is detected at diagnosis in approximately 15% of children with T-cell lymphoblastic lymphoma (T-LL). It is unclear whether the remaining patients have submicroscopic systemic disease and, if so, what is the clinical significance of this finding. PATIENTS AND METHODS Using a flow cytometric method that can detect one T-LL cell among 10,000 normal cells, we examined bone marrow and peripheral-blood samples collected from 99 children with T-LL at diagnosis, as well as blood samples collected from 42 patients during treatment. Results In 71 (71.7%) of the 99 marrow samples obtained at diagnosis, T-LL cells represented 0.01% to 31.6% (median, 0.22%) of mononuclear cells; 57 of the 71 T-LL-positive samples were from patients with stage II/III disease. Results of studies in bilateral marrow aspirates were highly concordant. Two-year event-free survival (EFS) was 68.1% +/- 11.1% (SE) for patients with > or = 1% T-LL cells in bone marrow versus 90.7% +/- 4.4% for those with lower levels of marrow involvement (P = .031); EFS for patients with > or = 5% lymphoblasts was 51.9% +/- 18.0% (P = .009). T-LL cells were as prevalent in blood as in marrow; monitoring residual T-LL cells in blood during remission induction therapy identified patients with slower disease clearance. CONCLUSION More than two thirds of children with T-LL have disseminated disease at diagnosis, a proportion much higher than previously demonstrated. Measurements of disease dissemination at diagnosis might provide useful prognostic information, which can be further refined by monitoring response to therapy through blood testing. PMID- 19546404 TI - Quality of life after pelvic radiotherapy or vaginal brachytherapy for endometrial cancer: first results of the randomized PORTEC-2 trial. AB - PURPOSE Studies on quality of life (QOL) among women with endometrial cancer have shown that patients who undergo pelvic radiotherapy report lower role functioning and more diarrhea and fatigue. In the Post Operative Radiation Therapy in Endometrial Cancer (PORTEC) trial, patients with endometrial carcinoma were randomly assigned to receive external-beam radiotherapy (EBRT) or vaginal brachytherapy (VBT). QOL was evaluated by using the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire C30 and subscales from the prostate cancer module, PR-25, and the ovarian cancer module, OV-28. PATIENTS AND METHODS PORTEC-2 accrued 427 patients between 2002 and 2006, of whom 214 were randomly assigned to EBRT, and 213 were randomly assigned to VBT. Three hundred forty-eight patients (81%) were evaluable for QOL. QOL outcomes were analyzed at a median follow-up of 2 years. Results At baseline after surgery, patient functioning was at the lowest level, and it increased during and after radiotherapy to reach a plateau after 12 months. Patients in the VBT group reported better social functioning (P < .002) and lower symptom scores for diarrhea, fecal leakage, the need to stay close to the toilet, and limitation in daily activities because of bowel symptoms (P < .001). At baseline, 15% of patients were sexually active; this increased significantly to 39% during the first year (P < .001). Sexual functioning and symptoms did not differ between the treatment groups. CONCLUSION Patients who received EBRT reported significantly higher levels of diarrhea and bowel symptoms. This resulted in a higher need to remain close to a toilet and, as a consequence, more limitation of daily activities because of bowel symptoms and decreased social functioning. Vaginal brachytherapy provides a better QOL, and should be the preferred treatment from a QOL perspective. PMID- 19546406 TI - Safety, pharmacokinetics, and antitumor activity of AMG 386, a selective angiopoietin inhibitor, in adult patients with advanced solid tumors. AB - PURPOSE AMG 386 is an investigational peptide-Fc fusion protein (ie, peptibody) that inhibits angiogenesis by preventing the interaction of angiopoietin-1 and angiopoietin-2 with their receptor, Tie2. This first-in-human study evaluated the safety, pharmacokinetics (PK), pharmacodynamics, and antitumor activity of AMG 386 in adults with advanced solid tumors. PATIENTS AND METHODS Patients in sequential cohorts received weekly intravenous AMG 386 doses of 0.3, 1, 3, 10, or 30 mg/kg. Results Thirty-two patients were enrolled on the study and received AMG 386. One occurrence of dose-limiting toxicity was seen at 30 mg/kg: respiratory arrest, which likely was caused by tumor burden that was possibly related to AMG 386. The most common toxicities were fatigue and peripheral edema. Proteinuria (n = 11) was observed without clinical sequelae. Only four patients (12%) experienced treatment-related toxicities greater than grade 1. A maximum tolerated dose was not reached. PK was dose-linear and the mean terminal-phase elimination half-life values ranged from 3.1 to 6.3 days. Serum AMG 386 levels appeared to reach steady-state after four weekly doses, and there was minimal accumulation. No anti-AMG 386 neutralizing antibodies were detected. Reductions in volume transfer constant (K(trans); measured by dynamic contrast-enhanced magnetic resonance imaging) were observed in 10 patients (13 lesions) 48 hours to 8 weeks after treatment. One patient with refractory ovarian cancer achieved a confirmed partial response (ie, 32.5% reduction by Response Evaluation Criteria in Solid Tumors) and withdrew from the study with a partial response after 156 weeks of treatment; four patients experienced stable disease for at least 16 weeks. CONCLUSION Weekly AMG 386 appeared well tolerated, and its safety profile appeared distinct from that of vascular endothelial growth factor-axis inhibitors. AMG 386 also appeared to impact tumor vascularity and showed antitumor activity in this patient population. PMID- 19546405 TI - Phase II study of thalidomide plus dexamethasone induction followed by tandem melphalan-based autotransplantation and thalidomide-plus-prednisone maintenance for untreated multiple myeloma: a southwest oncology group trial (S0204). AB - PURPOSE Thalidomide-dexamethasone (THAL-DEX) is standard induction therapy for multiple myeloma (MM). Tandem melphalan-based transplantations have yielded superior results to single transplantations. Phase II trial S0204 was designed to improve survival results reported for the predecessor, phase III trial S9321 by 50%. PATIENTS AND METHODS Newly diagnosed patients with MM were eligible for S0204 with THAL-DEX induction, tandem melphalan-based tandem transplantation, and THAL-prednisone maintenance. Results Of 143 eligible patients, 142 started induction, 73% completed first transplantation, 58% completed second transplantation, and 56% started maintenance. The quantity of stem cells required for two transplantations was reached in 88% of 111 patients undergoing collection, 74% of whom completed both transplantations. Partial response, very good partial remission, and complete response were documented after 12 months of maintenance therapy in 87%, 72%, and 22% of patients, respectively. During a median follow-up time of 37 months, 4-year estimates of event-free and overall survival were 50% and 64%, respectively. Survival outcomes were superior for International Staging System (ISS) stage 1 disease, when lactate dehydrogenase (LDH) levels were normal and a second transplantation was applied in a timely fashion. CONCLUSION Both overall survival (P = .0002) and event-free survival (P < .0001) were significantly improved with S0204 compared with S9321 when 121 and 363 patients, respectively, were matched on ISS stage and LDH. PMID- 19546407 TI - The silent dimension: expressing humanism in each medical encounter. PMID- 19546408 TI - Treating anemia with erythropoiesis-stimulating agents: effects on quality of life. PMID- 19546409 TI - Cost-effectiveness analysis of an established, effective procedure. PMID- 19546410 TI - The impact of selecting a high hemoglobin target level on health-related quality of life for patients with chronic kidney disease: a systematic review and meta analysis. AB - BACKGROUND: Treatment of anemia in chronic kidney disease (CKD) with erythropoietin-stimulating agents (ESAs) is commonplace. The optimal hemoglobin treatment target has not been established. A clearer understanding of the health related quality of life (HQOL) impact of hemoglobin target levels is needed. We systematically reviewed the randomized controlled trial (RCT) data on HQOL for patients treated with low to intermediate (9.0-12.0 g/dL) and high hemoglobin target levels (>12.0 g/dL) and performed a meta-analysis of all available 36-item short-form (SF-36) RCT data. METHODS: We conducted a search to identify all RCTs of ESA therapy in patients with anemia associated with CKD (1966-December 2006). Inclusion criteria were (1) 30 or more participants, (2) anemic adults with CKD, (3) epoetin (alfa and beta) or darbepoetin used, (4) a control arm, and (5) reported HQOL using a validated measure. All available SF-36 data underwent meta analysis using the weighted mean difference. RESULTS: Of 231 full texts screened, 11 eligible studies were identified. The SF-36 was used in 9 trials. Reporting of these data was generally incomplete. Data from each domain of the SF-36 were summarized. Statistically significant changes were noted in the physical function (weighted mean difference [WMD], 2.9; 95% confidence interval [CI], 1.3 to 4.5), general health (WMD, 2.7; 95% CI, 1.3 to 4.2), social function (WMD, 1.3; 95% CI, -0.8 to 3.4), and mental health (WMD, 0.4; 95% CI, 0.1 to 0.8) domains. None of the changes would be considered clinically significant. CONCLUSIONS: Our study suggests that targeting hemoglobin levels in excess of 12.0 g/dL leads to small and not clinically meaningful improvements in HQOL. This, in addition to significant safety concerns, suggests that targeting treatment to hemoglobin levels that are in the range of 9.0 to 12.0 g/dL is preferred. PMID- 19546413 TI - Frequency of failure to inform patients of clinically significant outpatient test results. AB - BACKGROUND: Failing to inform a patient of an abnormal outpatient test result can be a serious error, but little is known about the frequency of such errors or the processes for managing results that may reduce errors. METHODS: We conducted a retrospective medical record review of 5434 randomly selected patients aged 50 to 69 years in 19 community-based and 4 academic medical center primary care practices. Primary care practice physicians were surveyed about their processes for managing test results, and individual physicians were notified of apparent failures to inform and asked whether they had informed the patient. Blinded reviewers calculated a "process score" ranging from 0 to 5 for each practice using survey responses. RESULTS: The rate of apparent failures to inform or to document informing the patient was 7.1% (135 failures divided by 1889 abnormal results), with a range of 0% to 26.2%. The mean process score was 3.8 (range, 0.9 5.0). In mixed-effects logistic regression, higher process scores were associated with lower failure rates (odds ratio, 0.68; P < .001). Use of a "partial electronic medical record" (paper-based progress notes and electronic test results or vice versa) was associated with higher failure rates compared with not having an electronic medical record (odds ratio, 1.92; P = .03) or with having an electronic medical record that included both progress notes and test results (odds ratio, 2.37; P = .007). CONCLUSIONS: Failures to inform patients or to document informing patients of abnormal outpatient test results are common; use of simple processes for managing results is associated with lower failure rates. PMID- 19546411 TI - Cost-effectiveness of total knee arthroplasty in the United States: patient risk and hospital volume. AB - BACKGROUND: Total knee arthroplasty (TKA) relieves pain and improves quality of life for persons with advanced knee osteoarthritis. However, to our knowledge, the cost-effectiveness of TKA and the influences of hospital volume and patient risk on TKA cost-effectiveness have not been investigated in the United States. METHODS: We developed a Markov, state-transition, computer simulation model and populated it with Medicare claims data and cost and outcomes data from national and multinational sources. We projected lifetime costs and quality-adjusted life expectancy (QALE) for different risk populations and varied TKA intervention and hospital volume. Cost-effectiveness of TKA was estimated across all patient risk and hospital volume permutations. Finally, we conducted sensitivity analyses to determine various parameters' influences on cost-effectiveness. RESULTS: Overall, TKA increased QALE from 6.822 to 7.957 quality-adjusted life years (QALYs). Lifetime costs rose from $37,100 (no TKA) to $57 900 after TKA, resulting in an incremental cost-effectiveness ratio of $18,300 per QALY. For high-risk patients, TKA increased QALE from 5.713 to 6.594 QALY, yielding a cost-effectiveness ratio of $28,100 per QALY. At all risk levels, TKA was more costly and less effective in low-volume centers than in high-volume centers. Results were insensitive to variations of key input parameters within policy-relevant, clinically plausible ranges. The greatest variations were seen for the quality of life gain after TKA and the cost of TKA. CONCLUSIONS: Total knee arthroplasty appears to be cost effective in the US Medicare-aged population, as currently practiced across all risk groups. Policy decisions should be made on the basis of available local options for TKA. However, when a high-volume hospital is available, TKAs performed in a high-volume hospital confer even greater value per dollar spent than TKAs performed in low-volume centers. PMID- 19546414 TI - Cutaneous malignancies among HIV-infected persons. AB - BACKGROUND: As the life expectancy of persons infected with human immunodeficiency virus (HIV) increases, cancers have become an important cause of morbidity and mortality. Although cutaneous cancers are the most common malignant neoplasms in the general population, little data exist among HIV-positive persons, especially regarding the impact of HIV-specific factors. METHODS: We evaluated the incidence rates and factors associated with the development of cutaneous malignancies among HIV-infected persons by examining data that were prospectively collected from a large HIV study that included 4490 participants (1986-2006). Poisson regression and Cox proportional hazards models were performed. RESULTS: Six percent of HIV-infected persons (n = 254) developed a cutaneous malignancy during 33 760 person-years of follow-up (mean, 7.5 years). Since the advent of highly active antiretroviral therapy (HAART), the incidence rates of cutaneous non-AIDS-defining cancers (NADCs), in particular basal cell carcinoma, have exceeded the rates of cutaneous AIDS-defining cancers such as Kaposi sarcoma. Factors associated with the development of cutaneous NADCs in the multivariate models included increasing age (hazard ratio [HR], 2.1; 95% confidence interval [CI], 1.7-2.6) and race. Compared with the white/non-Hispanic race, African Americans (HR, 0.03; 95% CI, 0.01-0.14) and other races (HR, 0.14; 95% CI, 0.03-0.57) had a lower risk of cutaneous NADCs. There were no significant associations between cutaneous NADCs and time-updated CD4 lymphocyte counts, HIV RNA levels, or receipt of HAART. CONCLUSIONS: At present, the most common cutaneous malignancies among HIV-infected persons are NADCs. Cutaneous NADCs do not appear to be significantly associated with immune function or HAART but rather are related to traditional factors such as aging and skin color. PMID- 19546415 TI - Association between late-life social activity and motor decline in older adults. AB - BACKGROUND: Loss of motor function is a common consequence of aging, but little is known about the factors that predict idiopathic motor decline. Our objective was to test the hypothesis that late-life social activity is related to the rate of change in motor function in old age. METHODS: Longitudinal cohort study with a mean follow-up of 4.9 years with 906 persons without stroke, Parkinson disease, or dementia participating in the Rush Memory and Aging Project. At baseline, participants rated the frequency of their current participation in common social activities from which a summary measure of social activity was derived. The main outcome measure was annual change in a composite measure of global motor function, based on 9 measures of muscle strength and 9 motor performances. RESULTS: Mean (SD) social activity score at baseline was 2.6 (0.58), with higher scores indicating more frequent participation in social activities. In a generalized estimating equation model, controlling for age, sex, and education, global motor function declined by approximately 0.05 U/y (estimate, 0.016; 95% confidence interval [CI], -0.057 to 0.041 [P = .02]). Each 1-point decrease in social activity was associated with approximately a 33% more rapid rate of decline in motor function (estimate, 0.016; 95% CI, 0.003 to 0.029 [P = .02]). The effect of each 1-point decrease in the social activity score at baseline on the rate of change in global motor function was the same as being approximately 5 years older at baseline (age estimate, -0.003; 95% CI, -0.004 to -0.002 [P<.001]). Furthermore, this amount of motor decline per year was associated with a more than 40% increased risk of death (hazard ratio, 1.44; 95% CI, 1.30 to 1.60) and a 65% increased risk of incident Katz disability (hazard ratio, 1.65; 95% CI, 1.48 to 1.83). The association of social activity with the rate of global motor decline did not vary along demographic lines and was unchanged (estimate, 0.025; 95% CI, 0.005 to 0.045 [P = .01]) after controlling for potential confounders including late-life physical and cognitive activity, disability, global cognition depressive symptoms, body composition, and chronic medical conditions. CONCLUSION: Less frequent participation in social activities is associated with a more rapid rate of motor function decline in old age. PMID- 19546417 TI - The frequency of hyperkalemia and its significance in chronic kidney disease. AB - BACKGROUND: Hyperkalemia is a potential threat to patient safety in chronic kidney disease (CKD). This study determined the incidence of hyperkalemia in CKD and whether it is associated with excess mortality. METHODS: This retrospective analysis of a national cohort comprised 2 103 422 records from 245 808 veterans with at least 1 hospitalization and at least 1 inpatient or outpatient serum potassium record during the fiscal year 2005. Chronic kidney disease and treatment with angiotensin-converting enzyme inhibitors and/or angiotensin II receptor blockers (blockers of the renin-angiotensin-aldosterone system [RAAS]) were the key predictors of hyperkalemia. Death within 1 day of a hyperkalemic event was the principal outcome. RESULTS: Of the 66 259 hyperkalemic events (3.2% of records), more occurred as inpatient events (n = 34 937 [52.7%]) than as outpatient events (n = 31 322 [47.3%]). The adjusted rate of hyperkalemia was higher in patients with CKD than in those without CKD among individuals treated with RAAS blockers (7.67 vs 2.30 per 100 patient-months; P < .001) and those without RAAS blocker treatment (8.22 vs 1.77 per 100 patient-months; P < .001). The adjusted odds ratio (OR) of death with a moderate (potassium, >or=5.5 and <6.0 mEq/L [to convert to mmol/L, multiply by 1.0]) and severe (potassium, >or=6.0 mEq/L) hyperkalemic event was highest with no CKD (OR, 10.32 and 31.64, respectively) vs stage 3 (OR, 5.35 and 19.52, respectively), stage 4 (OR, 5.73 and 11.56, respectively), or stage 5 (OR, 2.31 and 8.02, respectively) CKD, with all P < .001 vs normokalemia and no CKD. CONCLUSIONS: The risk of hyperkalemia is increased with CKD, and its occurrence increases the odds of mortality within 1 day of the event. These findings underscore the importance of this metabolic disturbance as a threat to patient safety in CKD. PMID- 19546416 TI - Nocturnal Arrhythmias across a spectrum of obstructive and central sleep disordered breathing in older men: outcomes of sleep disorders in older men (MrOS sleep) study. AB - BACKGROUND: Rates of cardiac arrhythmias increase with age and may be associated with clinically significant morbidity. We studied the association between sleep disordered breathing (SDB) with nocturnal atrial fibrillation or flutter (AF) and complex ventricular ectopy (CVE) in older men. METHODS: A total of 2911 participants in the Outcomes of Sleep Disorders in Older Men Study underwent unattended polysomnography. Nocturnal AF and CVE were ascertained by electrocardiogram-specific analysis of the polysomnographic data. Exposures were (1) SDB defined by respiratory disturbance index (RDI) quartile (a major index including all apneas and hypopneas), and ancillary definitions incorporating (2) obstructive events, obstructive sleep apnea (OSA; Obstructive Apnea Hypopnea Index quartile), or (3) central events, central sleep apnea (CSA; Central Apnea Index category), and (4) hypoxia (percentage of sleep time with <90% arterial oxygen percent saturation). Multivariable logistic regression analyses were performed. RESULTS: An increasing RDI quartile was associated with increased odds of AF and CVE (P values for trend, .01 and <.001, respectively). The highest RDI quartile was associated with increased odds of AF (odds ratio [OR], 2.15; 95% confidence interval [CI], 1.19-3.89) and CVE (OR, 1.43; 95% CI, 1.12-1.82) compared with the lowest quartile. An increasing OSA quartile was significantly associated with increasing CVE (P value for trend, .01) but not AF. Central sleep apnea was more strongly associated with AF (OR, 2.69; 95% CI, 1.61-4.47) than CVE (OR, 1.27; 95% CI, 0.97-1.66). Hypoxia level was associated with CVE (P value for trend, <.001); those in the highest hypoxia category had an increased odds of CVE (OR, 1.62; 95% CI, 1.23-2.14) compared with the lowest quartile. CONCLUSIONS: In this large cohort of older men, increasing severity of SDB was associated with a progressive increase in odds of AF and CVE. When SDB was characterized according to central or obstructive subtypes, CVE was associated most strongly with OSA and hypoxia, whereas AF was most strongly associated with CSA, suggesting that different sleep-related stresses may contribute to atrial and ventricular arrhythmogenesis in older men. PMID- 19546418 TI - Low-molecular-weight heparin as an adjunct to thrombolysis in ST elevation myocardial infarction. PMID- 19546419 TI - Patient- and physician-oriented web sites and drug surveillance: bisphosphonates and severe bone, joint, and muscle pain. PMID- 19546420 TI - Cold-induced dehydration decreases tissue perfusion and increases blood pressure. PMID- 19546421 TI - Role of vitamin B12 in anemia in old age. PMID- 19546423 TI - Pitfalls in diagnosing chronic kidney disease from eGFR. PMID- 19546422 TI - Regarding the inverse relationship between blood pressure and outdoor temperature: it is the sun. PMID- 19546424 TI - Subclinical white matter integrity in subjects with cumulative lead exposure. AB - PURPOSE: To evaluate microstructural changes in the white matter of patients who were exposed to lead and to compare differences in fractional anisotropy (FA) between these patients and control subjects. MATERIALS AND METHODS: Institutional review board approval and subject informed consent were obtained for this HIPAA compliant study. Nineteen factory workers who had been exposed to lead and 18 healthy volunteers who had not were enrolled. FA values and T2-weighted fluid attenuation inversion-recovery magnetic resonance images were obtained at several regions of interest (the bilateral parietal, frontal, occipital, and temporal white matter and the genu and splenium of the corpus callosum). Lead levels were measured in the blood, midtibia, and patella. The Student t test was used to compare the difference in continuous variables between the two groups. Pearson correlation coefficients were used to assess the association between two variables. RESULTS: There were no significant differences in sex, age, body mass index, smoking history, betel nut consumption, or alcohol consumption between the factory workers and the volunteers. The number of milk drinkers among factory workers was significantly higher than that among volunteers (P < .001). The factory workers had significantly higher blood (P < .001), patella (P < .001), and midtibia (P = .005) lead levels than did the volunteers. Mean FA in the factory workers was lower than that in the volunteers at the same anatomic location; significant differences between the groups were noted bilaterally in the parietal, frontal, occipital, and temporal white matter. There was no significant difference in mean diffusivity values and mean T2 ratios between the factory workers and the volunteers. CONCLUSION: Decreased FA was associated with exposure to lead. Negative correlations between FA and blood, midtibia, and patella lead levels suggest that FA may be a useful index of early white matter damage. SUPPLEMENTAL MATERIAL: http://radiology.rsnajnls.org/cgi/content/full/2522080653/DC1. PMID- 19546425 TI - DeltaR2* in fetal sheep brains during hypoxia: MR imaging at 3.0 T versus that at 1.5 T. AB - PURPOSE: To investigate the feasibility of fetal blood oxygen level-dependent magnetic resonance (MR) imaging at 1.5 T and to compare DeltaR2* in the brains of fetal sheep during hypoxia at 3.0 T with that at 1.5 T. MATERIALS AND METHODS: All experimental protocols were reviewed and approved by the local authorities on animal protection. Between January 2006 and May 2006, fetal brain measurements were performed in eight pregnant ewes with 1.5-T and 3.0-T MR imaging units after fetal paralysis was achieved by administering pancuronium bromide. With both imaging units, a T2*-weighted single-shot gradient-echo echo-planar imaging sequence (echo time, 30 msec at 3.0 T and 50 msec at 1.5 T) was used to measure T2* signal changes (DeltaR2*) in the fetal brain in control conditions and during hypoxia (maternal oxygenation, 50%-70%). A carotid catheter was placed and maintained in the fetuses to enable measurement of the fetal arterial oxygen saturation (SaO(2)). DeltaR2* was correlated with fetal SaO(2), and linear regression analysis was performed. A paired t test was used to evaluate differences, with a significance level of P < .05. RESULTS: At both field strengths, a signal intensity decrease on T2*-weighted images during hypoxia was detected. At 1.5 T, mean fetal SaO(2) was reduced from 65.4% +/- 9.2 (standard deviation) during control conditions to 17.7% +/- 6.2 during hypoxia. DeltaR2* and fetal SaO(2) correlated significantly (r = 0.98, P = .018). At 3.0 T, fetal SaO(2) was reduced from 62.4% +/- 7.5 during control conditions to 18% +/- 7.5 during hypoxia. DeltaR2* and fetal SaO(2) also correlated significantly (r = 0.95, P = .012). A linear fit resulted in a slope value of 0.084 +/- 0.003 for 1.5 T and 0.166 +/- 0.016 for 3.0 T. This means a doubled sensitivity of DeltaR2* for oxygen saturation variations at 3.0 T compared with 1.5 T. CONCLUSION: MR imaging at 3.0 T is more sensitive than that at 1.5 T in the detection of DeltaR2* in the fetal brain during hypoxia. However, there was a signal decrease in the fetal brain in all 1.5-T experiments during hypoxia. Thus it is possible to measure fetal DeltaR2* at 1.5 T, which may be of more practical relevance for the evaluation of pregnant women. PMID- 19546426 TI - Coronary artery stenosis in high-risk patients: 64-section CT and coronary angiography--prospective study and analysis of discordance. AB - PURPOSE: To assess the diagnostic accuracy of multisection (64-section) computed tomography (CT) versus coronary angiography in detection of and assignment of grades for coronary artery stenoses in a high-risk population and to investigate causes for discordance between the two. MATERIALS AND METHODS: The protocol was approved by the local ethics committee. Patients gave informed consent. The study included 114 patients (103 men, 11 women; mean age, 63 years +/- 8.2 [standard deviation]) with potential myocardial ischemia. Multisection CT images were interpreted independently by two radiologists with unequal experience in reading coronary CT angiograms. Diagnostic performance of 64-section CT in detection of stenoses of 50% or more was assessed per patient, per artery, and per segment. Interrater agreement was assessed by using the Cohen kappa coefficient. Agreement between 64-section CT and coronary angiography for assigning grades to stenoses was assessed by using Bland-Altman analysis. RESULTS: Sixty-eight percent of patients had stenoses of 50% or more. Good interrater agreement was found, with kappa values of 0.77-0.85. For the most experienced radiologist, the sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio were 73.4%, 95.0%, 14.7, and 0.28 per segment, 95.2%, 94.7%, 18.0, and 0.05 per artery, and 100%, 89.2%, 9.26, and zero per patient, respectively. Discordance between 64 section CT and coronary angiography was related to either under- or overestimation of the degree of stenosis, anatomic misclassification, and coronary artery segments that were not assessable at 64-section CT. Bland-Altman analysis showed poor agreement, especially for intermediate stenosis (mean bias, 1.3%; 95% limits of agreement: -27.3%, 29.9%). CONCLUSION: Despite excellent sensitivity and negative likelihood ratios in a per-patient or per-vessel analysis, some coronary artery stenosis remained misdiagnosed with 64-section CT, resulting in limited sensitivity on a per-segment basis owing to anatomic discordance and failure to accurately quantify intermediate stenosis. PMID- 19546429 TI - US surveillance of regional lymph node recurrence after breast cancer surgery. AB - PURPOSE: To determine the diagnostic indexes of lymph node ultrasonography (US) of the axillary and supraclavicular regions for detecting lymph node recurrence (LNR) after breast cancer surgery and assess the effect of lymph node evaluation on prognosis during bilateral breast US. MATERIALS AND METHODS: Institutional review board approved this retrospective study and waived informed consent. Between January 2003 and December 2004, 3982 lymph node US examinations, including bilateral axillary and supraclavicular areas, were performed in 1817 women (mean age, 49.9 years; range, 22-86 years) after breast cancer surgery, nine of whom had palpable lesions. Final diagnosis was based on cytopathologic results, clinical follow-up, and imaging studies for at least 12 months after breast US. Diagnostic indexes of US for detecting LNR were assessed. The frequency of distant metastases between patients with ipsilateral LNR and those without was compared. Three-year mortality rates of patients with ipsilateral LNR only and those with distant metastases were evaluated. RESULTS: Of 1817 patients, 54 had suspicious LNR at US (28 at first, 20 at second, five at third, and one at fourth US examination). Thirty-nine of 1817 patients (2.1%), including nine with palpable lesions, had LNR, 11 of whom had ipsilateral LNR only. At first lymph node US, LNR was detected in 17 patients; at second, in 10; at third, in two; and at fourth, in one. Nine had false-negative results. The respective sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of lymph node US for detecting LNR per woman was 76.9% (30 of 39), 98.7% (1754 of 1778), 98.2% (1784 of 1817), 55.6% (30 of 54), and 99.5% (1754 of 1763); those of first lymph node US were 85.0% (17 of 20), 99.4% (1786 of 1797), 99.2% (1803 of 1817), 60.7% (17 of 28), and 99.8% (1786 of 1789); and those of total US examinations were 78.0% (32 of 41), 99.4% (3917 of 3941), 99.2% (3949 of 3982), 57.1% (32 of 56), and 99.8% (3917 of 3926). Distant metastases were found more frequently in patients with ipsilateral LNR (62%) than in those without (2.3%) (P < .0001). The 3-year mortality rate of patients with ipsilateral LNR only was significantly lower than that in patients with distant metastases (P = .03). CONCLUSION: Ipsilateral LNR is a predictor of distant metastasis, and lymph node evaluation during breast US is useful for early detection of LNR in asymptomatic patients. PMID- 19546428 TI - Carotid artery brain aneurysm model: in vivo molecular enzyme-specific MR imaging of active inflammation in a pilot study. AB - PURPOSE: To demonstrate the feasibility of using a myeloperoxidase (MPO)-specific paramagnetic magnetic resonance (MR) contrast agent to identify active inflammation in an animal model of common carotid artery (CCA) aneurysm. MATERIALS AND METHODS: All animal experiments were approved by the institutional animal care and use committee. Elastase-induced saccular aneurysms were created at the root of the right CCA in 16 New Zealand white rabbits. Intramural and perivascular injection of Escherichia coli lipopolysaccharide (LPS) was performed with an endovascular approach to induce aneurysm inflammation. After intraarterial injection of an MPO-specific (di-5-hydroxytryptamide of gadopentetate dimeglumine, 0.1 mmol per kilogram of bodyweight) or a non-MPO specific (di-tyrosine of gadopentetate dimeglumine, 0.1 mmol/kg) contrast agent, animals underwent 3-T MR imaging. Intramural presence of MPO in aneurysms in which LPS had been injected was confirmed at immunohistologic analysis. Active MPO activity was verified by measuring the spectrophotometric oxidation of guaiacol. RESULTS: Endovascular injection of LPS resulted in inflammatory cell infiltration into the aneurysm wall, and there was a difference in active MPO expression between aneurysms in which LPS had been injected and control aneurysms (20.3 ng of MPO per milligram of tissue vs 0.12 ng of MPO per milligram of tissue, respectively; P < .002). MR imaging with di-5-hydroxytryptamide of gadopentetate dimeglumine revealed a difference in enhancement ratio between inflamed aneurysms in which LPS had been injected and control aneurysms (1.55 +/- 0.05 vs 1.16 +/- 0.10, respectively; P < .02). In inflamed aneurysms, di-5 hydroxytryptamide of gadopentetate dimeglumine exhibited delayed washout kinetics compared with the kinetics of di-tyrosine of gadopentetate dimeglumine. This finding enabled the verification of MPO specificity. CONCLUSION: The findings of this pilot study established the feasibility of an animal model of saccular aneurysm inflammation that can be seen with clinical-field-strength MR imaging and use of the enzyme-sensitive MR contrast agent di-5-hydroxytryptamide of gadopentetate dimeglumine, which is a paramagnetic MPO substrate that specifically enhances MR signal. PMID- 19546430 TI - Measurement of hepatic lipid: high-speed T2-corrected multiecho acquisition at 1H MR spectroscopy--a rapid and accurate technique. AB - PURPOSE: To evaluate the feasibility, accuracy, and reproducibility of a fast breath-hold magnetic resonance (MR) spectroscopic method for T2-corrected hepatic lipid measurement in phantoms and in humans. MATERIALS AND METHODS: All experiments were institutional review board approved and HIPAA compliant; informed consent was obtained from all subjects. The 15-second breath-hold high speed T2-corrected multiecho (HISTO) MR spectroscopic technique was developed to acquire multiple echoes in a single acquisition, which enables the quantification of water and lipid T2, and subsequently to provide a corrected measure of hepatic lipid fraction. The accuracy of T2-corrected MR spectroscopy was evaluated in eight lipid phantoms doped with iron to simulate variable T2 effects. The mean absolute error of the HISTO technique with the known lipid amounts, as well as with uncorrected MR spectroscopic measures, was evaluated. The HISTO sequence was performed in 25 male subjects (mean age, 23.0 years +/- 19.2 [standard deviation]) to evaluate measurement bias with conventional, uncorrected MR spectroscopy. Three additional male subjects (mean age, 30.0 years +/- 1.0) were examined to assess reproducibility by using analysis of variance testing within subject and between separate imaging sessions. RESULTS: The absolute error in quantifying lipid fraction by using iron-doped lipid phantoms was less than 11% for the HISTO technique, compared with more than 50% for uncorrected MR spectroscopy. In the 25 human subjects, hepatic lipid measured by using HISTO differed significantly from that by using uncorrected MR spectroscopic methods by 5.1% +/- 2.6. Analysis of variance of three separate imaging sessions with the HISTO technique indicated no significant variance (P = .13) in three subjects. CONCLUSION: HISTO is an accurate, reproducible MR spectroscopic sequence for quantifying hepatic lipid noninvasively. Evidence has shown this method to be feasible in vivo for clinical use. PMID- 19546431 TI - Re: Promoting regular mammography screening I. A systematic assessment of validity in a randomized trial. PMID- 19546432 TI - RE: Moderate alcohol intake and cancer incidence in women. PMID- 19546433 TI - Re: Insulin, insulin-like growth factor-I, and risk of breast cancer in postmenopausal women. PMID- 19546434 TI - Severe intrauterine anemia: a new form of epsilongammagammadeltabeta thalassemia presenting in utero in a Norwegian family. AB - Severe intrauterine anemia of unknown cause presents a diagnostic challenge. We describe a Norwegian case, managed successfully by intrauterine transfusions, that further investigations demonstrated to be due to a rare type of thalassemia. A deletion of the 5' end of the beta globin gene cluster was characterized, the breakpoints sequenced and a new type of epsilongammagammadeltabeta thalassemia identified. This case highlights the need to consider diagnoses of rare conditions not normally associated with a particular population. PMID- 19546435 TI - Risk of solid tumors and myeloid hematological malignancies among first-degree relatives of patients with monoclonal gammopathy of undetermined significance. PMID- 19546436 TI - Mesenchymal stem cells efficiently inhibit the proinflammatory properties of 6 sulfo LacNAc dendritic cells. AB - Mesenchymal stem cells emerged as a promising treatment modality for steroid refractory graft-versus-host disease, which represents a major complication of allogeneic hematopoietic stem cell transplantation. Dendritic cells (DCs) display an extraordinary capacity to induce T-cell responses and play a crucial role in the pathogenesis of graft-versus-host disease. Here, we investigated the impact of mesenchymal stem cells on the proinflammatory capacity of 6-sulfo LacNAc (slan) dendritic cells, representing a major subpopulation of human blood dendritic cells. Mesenchymal stem cells markedly impair maturation of slanDCs and their ability to secrete proinflammatory cytokines, which was dependent on prostaglandin E(2). In contrast, the release of anti-inflammatory IL-10 was improved by mesenchymal stem cells. Furthermore, mesenchymal stem cells efficiently inhibit slanDC-induced proliferation of CD4(+) and CD8(+) T cells and polarization of naive CD4(+) T lymphocytes into Th1 cells. These results indicate that mesenchymal stem cells significantly impair the high proinflammatory capacity of slanDCs and further substantiate their potential for the treatment of graft-versus-host disease. PMID- 19546438 TI - Longitudinal growth retardation in a prepuberal girl with chronic myeloid leukemia on long-term treatment with imatinib. PMID- 19546441 TI - China's next great leap forward. PMID- 19546437 TI - Standardization of flow cytometry in myelodysplastic syndromes: report from the first European LeukemiaNet working conference on flow cytometry in myelodysplastic syndromes. AB - The myelodysplastic syndromes are a group of clonal hematopoietic stem cell diseases characterized by cytopenia(s), dysplasia in one or more cell lineages and increased risk of evolution to acute myeloid leukemia (AML). Recent advances in immunophenotyping of hematopoietic progenitor and maturing cells in dysplastic bone marrow point to a useful role for multiparameter flow cytometry (FCM) in the diagnosis and prognostication of myelodysplastic syndromes. In March 2008, representatives from 18 European institutes participated in a European LeukemiaNet (ELN) workshop held in Amsterdam as a first step towards standardization of FCM in myelodysplastic syndromes. Consensus was reached regarding standard methods for cell sampling, handling and processing. The group also defined minimal combinations of antibodies to analyze aberrant immunophenotypes and thus dysplasia. Examples are altered numbers of CD34(+) precursors, aberrant expression of markers on myeloblasts, maturing myeloid cells, monocytes or erythroid precursors and the expression of lineage infidelity markers. When applied in practice, aberrant FCM patterns correlate well with morphology, the subclassification of myelodysplastic syndromes, and prognostic scoring systems. However, the group also concluded that despite strong evidence for an impact of FCM in myelodysplastic syndromes, further (prospective) validation of markers and immunophenotypic patterns are required against control patient groups as well as further standardization in multi-center studies. Standardization of FCM in myelodysplastic syndromes may thus contribute to improved diagnosis and prognostication of myelodysplastic syndromes in the future. PMID- 19546439 TI - Diagnostic utility of flow cytometry in low-grade myelodysplastic syndromes: a prospective validation study. AB - BACKGROUND: The diagnosis of myelodysplastic syndromes is not always straightforward when patients lack specific diagnostic markers, such as blast excess, karyotype abnormality, and ringed sideroblasts. DESIGN AND METHODS: We designed a flow cytometry protocol applicable in many laboratories and verified its diagnostic utility in patients without those diagnostic markers. The cardinal parameters, analyzable from one cell aliquot, were myeloblasts (%), B-cell progenitors (%), myeloblast CD45 expression, and channel number of side scatter where the maximum number of granulocytes occurs. The adjunctive parameters were CD11b, CD15, and CD56 expression (%) on myeloblasts. Marrow samples from 106 control patients with cytopenia and 134 low-grade myelodysplastic syndromes patients, including 81 lacking both ringed sideroblasts and cytogenetic aberrations, were prospectively analyzed in Japan and Italy. RESULTS: Data outside the predetermined reference range in 2 or more parameters (multiple abnormalities) were common in myelodysplastic syndromes patients. In those lacking ringed sideroblasts and cytogenetic aberrations, multiple abnormalities were observed in 8/26 Japanese (30.8%) and 37/55 Italians (67.3%) when the cardinal parameters alone were considered, and in 17/26 Japanese (65.4%) and 42/47 Italians (89.4%) when all parameters were taken into account. Multiple abnormalities were rare in controls. When data from all parameters were used, the diagnostic sensitivities were 65% and 89%, specificities were 98% and 90%, and likelihood ratios were 28.1 and 8.5 for the Japanese and Italian cohorts, respectively. CONCLUSIONS: This protocol can be used in the diagnostic work-up of low-grade myelodysplastic syndromes patients who lack specific diagnostic markers, although further improvement in diagnostic power is desirable. PMID- 19546442 TI - Quebec left with numerous vacancies after resident match. PMID- 19546443 TI - Canada's genetic heterogeneity an asset in cord blood banking. PMID- 19546446 TI - Helping smokers with cardiac disease to abstain from tobacco after a stay in hospital. PMID- 19546444 TI - National trends in rates of death and hospital admissions related to acute myocardial infarction, heart failure and stroke, 1994-2004. AB - BACKGROUND: Rates of death from cardiovascular and cerebrovascular diseases have been steadily declining over the past few decades. Whether such declines are occurring to a similar degree for common disorders such as acute myocardial infarction, heart failure and stroke is uncertain. We examined recent national trends in mortality and rates of hospital admission for these 3 conditions. METHODS: We analyzed mortality data from Statistic Canada's Canadian Mortality Database and data on hospital admissions from the Canadian Institute for Health Information's Hospital Morbidity Database for the period 1994-2004. We determined age- and sex-standardized rates of death and hospital admissions per 100,000 population aged 20 years and over as well as in-hospital case-fatality rates. RESULTS: The overall age- and sex-standardized rate of death from cardiovascular disease in Canada declined 30.0%, from 360.6 per 100,000 in 1994 to 252.5 per 100 000 in 2004. During the same period, the rate fell 38.1% for acute myocardial infarction, 23.5% for heart failure and 28.2% for stroke, with improvements observed across most age and sex groups. The age- and sex-standardized rate of hospital admissions decreased 27.6% for stroke and 27.2% for heart failure. The rate for acute myocardial infarction fell only 9.2%. In contrast, the relative decline in the in-hospital case-fatality rate was greatest for acute myocardial infarction (33.1%; p < 0.001). Much smaller relative improvements in case fatality rates were noted for heart failure (8.1%) and stroke (8.9%). INTERPRETATION: The rates of death and hospital admissions for acute myocardial infarction, heart failure and stroke in Canada changed at different rates over the 10-year study period. Awareness of these trends may guide future efforts for health promotion and health care planning and help to determine priorities for research and treatment. PMID- 19546447 TI - Trends in cardiovascular disease: are we winning the war? PMID- 19546448 TI - Pharmacoepidemiology of insulin initiation in diabetes care. PMID- 19546449 TI - Harp seal heart valves being tested for human use. PMID- 19546450 TI - Limited onboard amenities. PMID- 19546452 TI - Cord blood bank plans evolve with "glacial speed". PMID- 19546455 TI - Smoking cessation initiated during hospital stay for patients with coronary artery disease: a randomized controlled trial. AB - BACKGROUND: Programs for smoking cessation for cardiac patients are underused in Canada. We examined the efficacy of an intervention for smoking cessation for patients admitted to hospital for coronary artery bypass graft (CABG) or because of acute myocardial infarction (MI). METHODS: Nurses randomly assigned 276 sequential patients admitted because of acute MI or for CABG who met the inclusion criteria. Participants received an intensive or minimal smoking cessation intervention. The minimal intervention included advice from physicians and nurses and 2 pamphlets. The intensive intervention included the minimal intervention plus 60 minutes of bedside counselling, take-home materials and 7 nurse-initiated counselling calls for 2 months after discharge. The outcomes were point prevalence of abstinence at 3, 6 and 12 months after discharge. RESULTS: The 12-month self-reported rate of abstinence was 62% among patients in the intensive group and 46% among those in the minimal group (odds ratio [OR] 2.0, 95% confidence interval [CI] 1.2-3.1). Abstinence was confirmed for 54% of patients in the intensive group and 35% in the minimal group (OR 2.0, 95% CI 1.3 3.6). Abstinence was significantly lower among those who used pharmacotherapy than among those who did not (p < 0.001). Continuous 12-month abstinence was 57% in the intensive group and 39% in the minimal group (p < 0.01). It was significantly higher among patients admitted for CABG than among those admitted because of acute MI (p < 0.05). INTERPRETATION: Providing intensive programs for smoking cessation for patients admitted for CABG or because of acute MI could have a major impact on health and health care costs. PMID- 19546456 TI - Initiation of insulin therapy in elderly patients taking oral antidiabetes drugs. AB - BACKGROUND: We sought to estimate the rate of initiation of insulin therapy among elderly patients using oral anti-diabetes drugs and to identify the factors associated with this initiation. METHODS: We conducted a population-based cohort study involving people aged 66 or more years who were newly dispensed an oral antidiabetes drug. Individuals who had received acarbose or a thiazolidinedione were excluded. The rate of insulin initiation was calculated by use of the Kaplan Meier method. Factors associated with insulin initiation were identified by multivariable Cox regression analyses. RESULTS: In this cohort of 69,674 new users of oral antidiabetes drugs, insulin was initiated at rate of 9.7 cases per 1000 patient-years. Patients who had initially received an insulin secretagogue (rather than metformin), who were prescribed an oral antidiabetes drug by an endocrinologist or an internist, who received higher initial doses of an oral antidiabetes drug, who received oral corticosteroids, used glucometer strips, or were admitted to hospital in the year before initiation of oral antidiabetes therapy, or who received 16 or more medications were more likely than those without these characteristics to have insulin therapy initiated. In contrast, patients who received thiazides or who used up to 12 medications (v. none) were less likely to have insulin therapy initiated. INTERPRETATION: Several factors related to drugs and health services are associated with the initiation of insulin therapy in elderly patients receiving oral antidiabetes drugs. It is unclear whether these factors predict secondary failure of oral antidiabetes drugs or instead reflect better management of type 2 diabetes. PMID- 19546457 TI - Stereotactic radiation therapy for inoperable, early-stage non-small-cell lung cancer. PMID- 19546458 TI - Carcinoid syndrome. PMID- 19546459 TI - Lung cancer screening. PMID- 19546460 TI - Clear language. PMID- 19546461 TI - Clear language. PMID- 19546463 TI - Interpret results with caution. PMID- 19546464 TI - New powers for naturopaths. PMID- 19546465 TI - Avalanche fatalities. PMID- 19546466 TI - Lung cancer screening. PMID- 19546469 TI - A moment in time. PMID- 19546470 TI - Prosecuting for knowingly transmitting HIV is warranted. PMID- 19546471 TI - Immunolocalization of aquaporin-1, -5, and -7 in the avian testis and vas deferens. AB - Thirteen mammalian aquaporin (AQP) isoforms have been identified, and they have a unique tissue-specific pattern of expression. AQPs have been found in the reproductive system of both male and female humans, rats, and mice. However, tissue expression and cellular and subcellular localization of AQPs have been poorly investigated in the male reproductive system of birds. The localization of AQP subtypes (AQP1, 2, 3, 4, 5, 7, 8, 9, and 11) in the goose testis and vas deferens has been studied through immunohistochemistry and immunobloting. Interestingly, the testicular and deferential tissues were positive for AQP1, -5, and -7 but not the others. AQP1 immunoreactivity was detected in the capillary endothelial cells of testis and vas deferens. AQP5 was localized in the interstitial tissue of the testis, including Leydig cells, as well as in the basal cells of vas deferens. Double-labeling confocal microscopy revealed coexpression of AQP5 with capillary AQP1 in the testis. AQP7 was expressed in elongated spermatid and spermatozoa tails in the testis, as well as spermatozoa tails in the vas deferens. These results suggest that several subtypes of AQPs are involved in the regulation of water homeostasis in the goose male reproductive system. PMID- 19546472 TI - Advanced osteoarthritis in humans is associated with altered collagen VI expression and upregulation of ER-stress markers Grp78 and bag-1. AB - To test the hypothesis that a perturbation of endoplasmic reticulum (ER) function is involved in the pathogenesis of osteoarthritis (OA), articular cartilage was isolated from non-OA patients secondary to resection of osteo- or chondrosarcomas. Intra-joint samples of minimal and advanced osteoarthritic cartilage were isolated from patients undergoing total knee arthroplasty and scored for disease severity. Glucose-regulated protein-78 (grp78) and bcl-2 associated athanogene-1 (bag-1) were detected via immunofluorescence as markers of non-homeostatic ER function. Additionally, the expression of type VI collagen and its integrin receptor, NG2, was determined to examine cartilage matrix health and turnover. There was an upregulation of grp78 in advanced OA, and variable expression in minimal OA. Non-OA cartilage was consistently grp78 negative. The downstream regulator bag-1 was also upregulated in OA compared with normal cartilage. Collagen VI was mainly cell-associated in non-OA cartilage, with a more widespread distribution observed in OA cartilage along with increased intracellular staining intensity. The collagen VI integral membrane proteoglycan receptor NG2 was downregulated in advanced OA compared with its patient-matched minimally involved cartilage sample. These results suggest that chondrocytes exhibit ER stress during OA, in association with upregulation of a large secreted molecule, type VI collagen. PMID- 19546473 TI - Glucose-6-phosphate dehydrogenase deficiency and malaria: cytochemical detection of heterozygous G6PD deficiency in women. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a X-chromosomally transmitted disorder of the erythrocyte that affects 400 million people worldwide. Diagnosis of heterozygously-deficient women is complicated: as a result of lyonization, these women have a normal and a G6PD-deficient population of erythrocytes. The cytochemical assay is the only reliable assay to discriminate between heterozygously-deficient women and non-deficient women or homozygously-deficient women. G6PD deficiency is mainly found in areas where malaria is or has been endemic. In these areas, malaria is treated with drugs that can cause (severe) hemolysis in G6PD-deficient individuals. A cheap and reliable test is necessary for diagnosing the deficiency to prevent hemolytic disorders when treating malaria. In this review, it is concluded that the use of two different tests for diagnosing men and women is the ideal approach to detect G6PD deficiency. The fluorescent spot test is inexpensive and easy to perform but only reliable for discriminating hemizygous G6PD-deficient men from non-deficient men. For women, the cytochemical assay is recommended. However, this assay is more expensive and difficult to perform and should be simplified into a kit for use in developing countries. PMID- 19546474 TI - Bovine lactadherin as a calcium-independent imaging agent of phosphatidylserine expressed on the surface of apoptotic HeLa cells. AB - Bovine lactadherin holds a stereo-specific affinity for phosphatidylserine (PS) membrane domains and binds at PS concentrations lower than the benchmark PS probe, annexin V. Accordingly, lactadherin has recognized PS exposure on preapoptotic immortalized leukemia cells at an earlier time point than has annexin V. In the present study, the cervical cancer cell line HeLa has been employed as a model system to compare the topographic distribution of PS with the two PS binding proteins as adherent cells enter the apoptotic program. HeLa cells were cultured on glass-bottom Petri dishes, and apoptosis was induced by staurosporine. Fluorescence-labeled lactadherin and/or annexin V were used to detect PS exposure by confocal microscopy. Both lactadherin and annexin V staining revealed PS localized to plasma membrane rim and blebs. In addition, lactadherin identified PS exposure on long filopodia-like extensions, whereas annexin V internalized in granule-like structures. All in all, the data further delineate the differences in PS binding patterns of lactadherin and annexin V. PMID- 19546475 TI - Toll-like receptors 3, 4, and 7 are expressed in the enteric nervous system and dorsal root ganglia. AB - The aim of the present study was to evaluate the expression of innate immunity receptors belonging to the Toll-like family in the neural plexuses of the different tracts of murine intestine, of the human ileum, and in lower dorsal root ganglia (DRGs) from where extrinsic afferents to these plexuses originate. Results obtained by immunohistochemistry and immunofluorescence on paraffin embedded tissue and whole-mount preparations show that Toll-like receptors (TLRs) -3 and -7, recognizing viral RNA, and TLR4, recognizing lipopolysaccharide (membrane component of Gram-negative bacteria), are expressed in the myenteric and submucous plexuses of murine intestine and human ileum, and in DRGs primary sensory neurons. They also show that TLR4 immunostaining is stronger in murine distal large bowel. In murine tissue, expression of TLRs was present in both neurons and glial cells. These observations indicate that the enteric neural network might be directly activated by bacterial and viral components and is therefore more in the forefront than previously envisaged in defense responses of the intestinal wall and in the cross-talk with intestinal microbiota. They also highlight the presence of a peripheral neural network that by way of hardwired neurotransmission could potentially convey to the central nervous system specific information on our microbial counterpart and invading or potentially invading pathogens. PMID- 19546476 TI - Early epigenetic changes and DNA damage do not predict clinical response in an overlapping schedule of 5-azacytidine and entinostat in patients with myeloid malignancies. AB - Sequential administration of DNA methyltransferase (DNMT) inhibitors and histone deacetylase (HDAC) inhibitors has demonstrated clinical efficacy in patients with hematologic malignancies. However, the mechanism behind their clinical efficacy remains controversial. In this study, the methylation dynamics of 4 TSGs (p15(INK4B), CDH-1, DAPK-1, and SOCS-1) were studied in sequential bone marrow samples from 30 patients with myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML) who completed a minimum of 4 cycles of therapy with 5-azacytidine and entinostat. Reversal of promoter methylation after therapy was observed in both clinical responders and nonresponders across all genes. There was no association between clinical response and either baseline methylation or methylation reversal in the bone marrow or purified CD34(+) population, nor was there an association with change in gene expression. Transient global hypomethylation was observed in samples after treatment but was not associated with clinical response. Induction of histone H3/H4 acetylation and the DNA damage associated variant histone gamma-H2AX was observed in peripheral blood samples across all dose cohorts. In conclusion, methylation reversal of candidate TSGs during cycle 1 of therapy was not predictive of clinical response to combination "epigenetic" therapy. This trial is registered with http://www.clinicaltrials.gov under NCT00101179. PMID- 19546477 TI - A comparison of neurocognitive functioning in children previously randomized to dexamethasone or prednisone in the treatment of childhood acute lymphoblastic leukemia. AB - In previous clinical trials of childhood acute lymphoblastic leukemia (ALL), dexamethasone resulted in higher event-free survival rates than prednisone, presumably due to greater central nervous system penetration. Dexamethasone's association with long-term neurocognitive toxicity is unknown. In this multisite study, we measured neurocognitive functioning in 92 children with standard-risk ALL, 1 to 9.99 years at diagnosis, at a mean of 9.8 years after randomization to prednisone (n = 41) or dexamethasone (n = 51) on Children's Cancer Group (CCG) 1922. No significant overall differences in mean neurocognitive and academic performance scores were found between the prednisone and dexamethasone groups after adjusting for age, sex, and time since diagnosis. The exception was that patients receiving dexamethasone scored one-third of a standard deviation worse on word reading (98.8 +/- 1.7 vs 104.9 +/- 1.8; P = .02). There were no group differences in the distribution of test scores or the parents' report of neurologic complications, psychotropic drug use, and special education. Further analyses suggested for the dexamethasone group, older age of diagnosis was associated with worse neurocognitive functioning; for the prednisone group, younger age at diagnosis was associated with worse functioning. In conclusion, our study did not demonstrate any meaningful differences in long-term cognitive functioning of childhood ALL patients based on corticosteroid randomization. This study is registered with http://www.clinicaltrials.gov under NCT00085176. PMID- 19546480 TI - A confirmatory factor analysis of the California Verbal Learning Test--Second Edition (CVLT-II) in a traumatic brain injury sample. AB - The latent structure of the California Verbal Learning Test-Second Edition (CVLT II) was examined in a clinical sample of 223 persons with traumatic brain injury that had been screened to remove individuals with complicating premorbid (e.g., psychiatric) or comorbid (e.g., financial compensation seeking) histories. Analyses incorporated the z scores from 12 CVLT-II variables. Maximum-likelihood confirmatory analyses were performed to test the fit and parsimony of four hypothetical models. A four-factor model, consisting of Attention Span, Learning Efficiency, Delayed Memory, and Inaccurate Memory, met all the a priori specified criteria for model fit and parsimony. This model was consistent with that identified previously in a confirmatory factor analysis of the CVLT-II standardization sample. The results support the construct validity of the CVLT-II in individuals with traumatic brain injury and indicate that a multifactorial interpretation is appropriate for clinical practice. PMID- 19546478 TI - Direct crosstalk between mast cell-TNF and TNFR1-expressing endothelia mediates local tissue inflammation. AB - Signaling through tumor necrosis factor receptor 1 (TNFR1) controls bacterial infections and the induction of inflammatory Th1 cell-mediated autoimmune diseases. By dissecting Th1 cell-mediated delayed-type hypersensitivity responses (DTHRs) into single steps, we localized a central defect to the missing TNFR1 expression by endothelial cells (ECs). Adoptive transfer and mast cell knockin experiments into Kit(W)/Kit(W-v), TNF(-/-), and TNFR1(-/-) mice showed that the signaling defect exclusively affects mast cell-EC interactions but not T cells or antigen-presenting cells. As a consequence, TNFR1(-/-) mice had strongly reduced mRNA and protein expression of P-selectin, E-selectin, ICAM-1, and VCAM-1 during DTHR elicitation. In consequence, intravital fluorescence microscopy revealed up to 80% reduction of leukocyte rolling and firm adhesion in TNFR1(-/-) mice. As substitution of TNF(-/-) mice with TNF-producing mast cells fully restored DTHR in these mice, signaling of mast cell-derived TNF through TNFR1-expressing ECs is essential for the recruitment of leukocytes into sites of inflammation. PMID- 19546479 TI - Surface RANKL of Toll-like receptor 4-stimulated human neutrophils activates osteoclastic bone resorption. AB - Inflammatory bone loss in septic and inflammatory conditions is due to increased activity of osteoclasts that requires receptor activator of NF-kappa B-ligand (RANKL). Neutrophils are the predominant infiltrating cells in these conditions. Although disease severity is linked to neutrophils, their role in evolution of bony lesions is not clear. We show that lipopolysaccharide (LPS), a toll-like receptor 4 ligand, up-regulated the expression of membrane RANKL in human blood neutrophils and murine air pouch-derived neutrophils. LPS-activated human and murine neutrophils, cocultured with human monocyte-derived osteoclasts and RAW 264.7 cells, respectively, stimulated bone resorption. Transfection of PLB-985 neutrophil-like cells with RANKL antisense RNA reduced osteoclastogenesis. Synovial fluid neutrophils of patients with exacerbation of rheumatoid arthritis strongly expressed RANKL and activated osteoclastogenesis in coculture systems. Osteoprotegerin, the RANKL decoy receptor, suppressed osteoclast activation by neutrophils from these different sources. Moreover, direct cell-cell contact between neutrophils and osteoclasts was visualized by confocal laser microscopy. Activation of neutrophil membrane-bound RANKL was linked to tyrosine phosphorylation of Src-homology domain-containing cytosolic phosphatase 1 with concomitant down-regulation of cytokine production. The demonstration of these novel functions of neutrophils highlights their potential role in osteoimmunology and in therapeutics of inflammatory bone disease. PMID- 19546481 TI - The docking technique for medial patellofemoral ligament reconstruction: surgical technique and clinical outcome. AB - BACKGROUND: Current techniques of medial patellofemoral ligament (MPFL) reconstruction vary with respect to methods of fixation on the femur and the patella. This article presents the outcomes of a surgical technique for reconstruction of the MPFL that uses a soft tissue graft with interference screw fixation on the femur and a docking technique for fixation on the patella. HYPOTHESIS: Patients with patellar instability who are treated with the docking technique for MPFL reconstruction will have improvements in knee symptoms and function, with a high percentage achieving good to excellent results at early follow-up. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Twenty consecutive patients with patellar instability underwent reconstruction of the MPFL. Patients were evaluated preoperatively and postoperatively by physical and radiographic examination and subjectively with the IKDC (International Knee Documentation Committee), Tegner, Kujala, and Lysholm questionnaires. Nineteen patients underwent magnetic resonance imaging preoperatively. RESULTS: The average follow-up was 31 months (range, 24-39). No recurrent episodes of dislocation or subluxation were reported. A firm endpoint to lateral patellar translation was noted in all patients at most recent follow-up. The IKDC subjective knee evaluation score improved from 42 preoperatively to 82 postoperatively (P < .001); Kujala, from 50 to 88 (P < .001); Lysholm, from 50 to 89 (P < .001); and Tegner, from 3.6 to 5.6 (P < .001). CONCLUSION: The docking technique for MPFL reconstruction is an effective surgical procedure for the treatment of patellar instability. PMID- 19546482 TI - Distal biceps tendon rupture: a new repair technique in 14 patients using the biotenodesis screw. AB - BACKGROUND: Distal biceps tendon ruptures are uncommon injuries. Operative treatment has been shown to improve functional outcomes. A variety of surgical repair techniques have been described for distal biceps ruptures. PURPOSE: The authors present their experience with a new technique to anatomically repair distal biceps tendon ruptures through a single-incision approach that they believe is a safe and reliable method of achieving repair. STUDY DESIGN: Case series; Level of evidence, 4. MATERIALS AND METHODS: Fourteen patients with 14 biceps tendon ruptures underwent a repair with a bioabsorbable Biotenodesis screw. All 14 patients underwent clinical assessment using the Mayo Elbow Performance Score, measurement of range of motion, and flexion strength testing. Mean follow-up was 29.1 months. RESULTS: Three patients had a good result and 11 patients had an excellent result. The mean elbow flexion arc was 141.4 degrees (range, 125 degrees -155 degrees ; standard deviation, 7.19 degrees ) with no flexion contractures in the operated side compared with the unaffected elbow. All patients achieved an equal range of pronation/supination to the unaffected side. The mean flexion strength in the injured arm was 25.7 kg, compared with 26.9 kg in the uninjured side. No complications were noted about the elbow. CONCLUSION: The authors believe this new technique gives a good functional outcome with reproducible results. PMID- 19546483 TI - Archetype-based electronic health records: a literature review and evaluation of their applicability to health data interoperability and access. AB - Health Information Managers (HIMs) are responsible for overseeing health information. The change management necessary during the transition to electronic health records (EHR) is substantial, and ongoing. Archetype-based EHRs are a core health information system component which solve many of the problems that arise during this period of change. Archetypes are models of clinical content, and they have many beneficial properties. They are interoperable, both between settings and through time. They are more amenable to change than conventional paradigms, and their design is congruent with clinical practice. This paper is an overview of the current archetype literature relevant to Health Information Managers. The literature was sourced in the English language sections of ScienceDirect, IEEE Explore, Pubmed, Google Scholar, ACM Digital library and other databases on the usage of archetypes for electronic health record storage, looking at the current areas of archetype research, appropriate usage, and future research. We also used reference lists from the cited papers, papers referenced by the openEHR website, and the recommendations from experts in the area. Criteria for inclusion were (a) if studies covered archetype research and (b) were either studies of archetype use, archetype system design, or archetype effectiveness. The 47 papers included show a wide and increasing worldwide archetype usage, in a variety of medical domains. Most of the papers noted that archetypes are an appropriate solution for future-proof and interoperable medical data storage. We conclude that archetypes are a suitable solution for the complex problem of electronic health record storage and interoperability. PMID- 19546484 TI - The preparedness of hospital Health Information Services for system failures due to internal disasters. AB - The unimpeded functioning of hospital Health Information Services (HIS) is essential for patient care, clinical governance, organisational performance measurement, funding and research. In an investigation of hospital Health Information Services' preparedness for internal disasters, all hospitals in the state of Victoria with the following characteristics were surveyed: they have a Health Information Service/ Department; there is a Manager of the Health Information Service/Department; and their inpatient capacity is greater than 80 beds. Fifty percent of the respondents have experienced an internal disaster within the past decade, the majority affecting the Health Information Service. The most commonly occurring internal disasters were computer system failure and floods. Two-thirds of the hospitals have internal disaster plans; the most frequently occurring scenarios provided for are computer system failure, power failure and fire. More large hospitals have established back-up systems than medium- and small-size hospitals. Fifty-three percent of hospitals have a recovery plan for internal disasters. Hospitals typically self-rate as having a 'medium' level of internal disaster preparedness. Overall, large hospitals are better prepared for internal disasters than medium and small hospitals, and preparation for disruption of computer systems and medical record services is relatively high on their agendas. PMID- 19546485 TI - Transformation of Emergency Department processes of care with EHR, CPOE, and ER event tracking systems. AB - Mercy Medical Center, North Iowa implemented electronic health records (EHR), computerised provider order entry (CPOE) and event tracking systems in the emergency department (ED) as part of hospital-wide implementation of clinical information systems. This case study examines the changes in outcomes and processes in the ED following implementation. Although the system was designed to enhance efficiency, there was a significant increase in the mean length of stay (about 17 minutes, or 15%) in the ED after implementation. This surprising finding was examined in relationship to the multiple process-of-care changes in the ED. PMID- 19546486 TI - Users' attitudes to an electronic medical record system and its correlates: a multivariate analysis. AB - Implementation of an electronic medical record (EMR) system increases efficiency of health services, quality of care and patient satisfaction. Successful implementation depends on many factors, one of which is how users respond to the new system. We studied medical receptionists' appraisal of the newly implemented EMR system in primary healthcare centres in Kuwait. Four hundred receptionists were selected randomly from different healthcare centres and asked to complete a user interaction satisfaction questionnaire relating to their experience of the new system. The response rate was 80.5%. A large majority of the respondents considered the system to be flexible (83%), easy (89%), and satisfying (81%). However, more than one third of the respondents (36%) found the system inadequate. Bivariate and multivariate analyses found age, typing ability, ease of data entry and computer error as significant correlates with overall user response. These findings relating to users' reactions to various aspects of the EMR should assist policymakers to recognise the causes of dissatisfaction with the EMR among medical receptionists at health centre clinics that may adversely affect its successful implementation and regular use, as well as the quality of care provided by the clinics. In addition, the findings provide information to assist the development of guidelines for future implementation of the EMR system at the secondary healthcare level. PMID- 19546488 TI - The fabrication of periodic metal nanodot arrays through pulsed laser melting induced fragmentation of metal nanogratings. AB - We propose and demonstrate a new method for fabricating periodic arrays of metal nanodots over a large area. In this method, metal nanogratings were first patterned on a substrate by nanoimprint lithography and lift-off, and were then melted using a single excimer laser pulse. We found that the laser melting broke the metal nanogratings into periodic nanodots. Furthermore, we demonstrated the control of the nanodot array pitch using a substrate surface topology, and the fabrication of two-dimensional periodic metal nanodot arrays of 70 nm diameter and 200 nm period. The fragmentation of lines into dots was attributed to the Rayleigh instability in a liquid cylinder. PMID- 19546489 TI - The fabrication of carbon nanotube probes utilizing ultra-high vacuum transmission electron microscopy. AB - An application of ultra-high vacuum transmission electron microscopy (UHV TEM) is demonstrated for the fabrication of carbon nanotube (CNT) probes. In this study, all the fabrication processes -- such as CNT attachment, CNT orientation manipulation, and apex trimming -- are integrated into a single UHV TEM system. The in situ work under UHV conditions (<5 x 10(-10) mbar) allows us to clean the tip surface at the start of the fabrication process to ensure a good contact between the tip and CNT. Furthermore, the CNT size can be user-selected to meet the various needs for scanning probe microscopy (SPM). Most importantly, the in situ trimming enables a multi-walled CNT to have the sharpness of a single-walled CNT. The three advantages mentioned above are designed to improve conventional methods and will be shown in detail as the procedures of CNT probe fabrication by a series of high-resolution TEM images. Finally, we compare the scanned image via our CNT probes and conventional probes, where the typical artefacts coming from the conventional ones are addressed. We believe the technique we have developed here will further enhance the resolution of SPM measurements. PMID- 19546490 TI - The plasmonic engineering of metal nanoparticles for enhanced fluorescence and Raman scattering. AB - We have investigated the effects of tuning the localized surface plasmon resonances (LSPRs) of silver nanoparticles on the fluorescence intensity, lifetime, and Raman signal from nearby fluorophores. The presence of a metallic structure can alter the optical properties of a molecule by increasing the excitation field, and by modifying radiative and nonradiative decay mechanisms. By careful choice of experimental parameters we have been able to decouple these effects. We observe a fourfold increase in fluorescence enhancement and an almost 30-fold increase in decay rate from arrays of Ag nanoparticles, when the LSPR is tuned to the emission wavelength of a locally situated fluorophore. This is consistent with a greatly increased efficiency for energy transfer from fluorophores to surface plasmons, resulting in a significant increase in quantum yield. Additionally, spatial mapping of the surface enhanced Raman scattering signal from a nanoparticle array reveals highly localized differences in the excitation field. PMID- 19546491 TI - Cell nanomechanics and focal adhesions are regulated by retinol and conjugated linoleic acid in a dose-dependent manner. AB - Retinol and conjugated linoleic acid (CLA) have previously been shown to have an important role in gene expression and various cellular processes, including differentiation, proliferation and cell death. In this study we have investigated the effect of retinol and CLA, both individually and in combination, on the intracellular cytoskeleton, focal adhesions (FAs) and the nanomechanical properties of 3T3 fibroblasts. We observed a dose-dependent decrease in the formation of FAs following treatment with either compound, which was directly correlated to an increase in cell height (>30%) and a decrease in the measured Young's modulus (approximately 28%). Furthermore, treatments with both compounds demonstrated an increased effect and led to a reduction of >70% in the average number of FAs per cell and a decrease of >50% in average cell stiffness. These data reveal that retinol and CLA disrupt FA formation, leading to an increase in cell height and a significant decrease in stiffness. These results may broaden our understanding of the interplay between cell nanomechanics and cellular contact with the external microenvironment, and help to shed light on the important role of retinoids and CLA in health and disease. PMID- 19546492 TI - Contacting individual Fe(110) dots in a single electron-beam lithography step. AB - We report on a new approach, entirely based on an electron-beam lithography technique, to contact electrically, in a four-probe scheme, single nanostructures obtained by self-assembly. In our procedure, nanostructures of interest are located and contacted in the same fabrication step. This technique has been developed to study the field-induced reversal of an internal component of an asymmetric Bloch domain wall observed in elongated structures such as Fe(110) dots. We have focused on the control, using an external magnetic field, of the magnetization orientation within Neel caps that terminate the domain wall at both interfaces. Preliminary magneto-transport measurements are discussed demonstrating that single Fe(110) dots have been contacted. PMID- 19546493 TI - Multi-walled carbon nanotubes suppress potassium channel activities in PC12 cells. AB - The advancement in nanotechnology has produced technological and conceptual breakthroughs but the effects nanomaterials have on organisms at the cellular level are poorly understood. Here we report that carboxyl-terminated multi-walled carbon nanotubes (MWCNTs) act as antagonists of three types of potassium channels as assessed by whole-cell patch clamp electrophysiology on undifferentiated pheochromocytoma (PC12) cells. Our results showed that carboxyl-terminated MWCNTs suppress the current densities of I(to), I(K) and I(K1) in a time-dependent and irreversible manner. The suppressions were most distinct 24 h after incubation with MWCNTs. However, MWCNTs did not significantly change the expression levels of reactive oxygen species (ROS) or intracellular free calcium and also did not alter the mitochondrial membrane potential (DeltaPsi(m)) in PC12 cells. These results suggest that oxidative stress was not involved in the MWCNTs suppression of I(to), I(K) and I(K1) current densities. Nonetheless, the suppression of potassium currents by MWCNTs will impact on electrical signaling of excitable cells such as neurons and muscles. PMID- 19546494 TI - Filling of hole arrays with InAs quantum dots. AB - Focused ion beams are used to pattern GaAs(001) surfaces with an array of nanometer-deep holes upon which deposition of InAs results in quantum dot formation at the hole location. Experiments show that the size and quantity of quantum dots formed depend on growth parameters, and ion dose, which affects the size and shape of the resulting holes. Quantum dots fabricated in this fashion have a photoluminescence peak at 1.28 eV at 77 K, indicating that the ion irradiation due to patterning does not destroy their optical activity. Kinetic Monte Carlo simulations that include elastic relaxation qualitatively model the growth of dots in nanometer-deep holes, and demonstrate that growth temperature, depth of the holes, and the angle of the hole sidewalls strongly influence the number of quantum dots that form at their perimeter. PMID- 19546495 TI - Dual patterning and sequential functionalization of block copolymers using photocrosslinkable random copolymer film. AB - A novel method of dual patterning and sequential functionalization of block copolymers is proposed using a photocrosslinkable random copolymer film, poly(styrene-r-(tert-butyl acrylate)-r-(cinnamoyloxyethyl acrylate)). The tert butyl esters of the block copolymer, polystyrene-block-poly(tert-butyl acrylate), coated on the patterned random copolymer film were sequentially deprotected to give carboxylic acids using an acid catalyst and heat treatment. The sequentially produced carboxylic acid patterns were used for the sequential patterning of gold nanoparticles as an example of potential applications. PMID- 19546496 TI - An angled nano-tunnel fabricated on poly(methyl methacrylate) by a focused ion beam. AB - Angled nano-scale tunnels with high aspect ratio were fabricated on poly(methyl methacrylate) (PMMA) using a focused ion beam (FIB). The fabrication parameters such as ion fluence, incidence angle, and acceleration voltage of the Ga(+) ion beam were first studied on the PMMA surface to explore the formation of the nano scale configurations such as nano-holes and cones with diameter in the range of 50-150 nm at an ion beam acceleration voltage of 5-20 kV. It was also found that the PMMA surface exposed to FIB was changed into an amorphous graphitic structure. Angled nano-scale tunnels were fabricated with high aspect ratio of 700-1500 nm in depth and 60 nm in mean diameter at an ion beam acceleration voltage of 5 kV and under a specific ion beam current. The angle of the nano tunnels was found to follow the incident angle of the ion beam tilted from 0 degrees to 85 degrees , which has the potential for creating a mold for anisotropic adhesives by mimicking the hairs on a gecko's feet. PMID- 19546497 TI - The fabrication of plasmonic Au nanovoid trench arrays by guided self-assembly. AB - Metallic nanoscale voids can support both localized and propagating plasmons and form plasmonic crystals. However, constructing 1D arrays is crucial for producing plasmonic circuits. In this paper we report the first experimental evidence of plasmons in templated linear arrays of self-assembled structures. Single and multilayer arrays of gold voids have been fabricated by self-assembly of sub micron polystyrene spheres in V-shaped trenches in silicon, followed by selective area electrodeposition. Angle-dependent dispersion characteristics reveal the existence of localized plasmons. PMID- 19546498 TI - Size dependence in tunneling spectra of PbSe quantum-dot arrays. AB - Interdot Coulomb interactions and collective Coulomb blockade were theoretically argued to be a newly important topic, and experimentally identified in semiconductor quantum dots, formed in the gate confined two-dimensional electron gas system. Developments of cluster science and colloidal synthesis accelerated the studies of electron transport in colloidal nanocrystal or quantum-dot solids. To study the interdot coupling, various sizes of two-dimensional arrays of colloidal PbSe quantum dots are self-assembled on flat gold surfaces for scanning tunneling microscopy and scanning tunneling spectroscopy measurements at both room and liquid-nitrogen temperatures. The tip-to-array, array-to-substrate, and interdot capacitances are evaluated and the tunneling spectra of quantum-dot arrays are analyzed by the theory of collective Coulomb blockade. The current voltage of PbSe quantum-dot arrays conforms properly to a scaling power law function. In this study, the dependence of tunneling spectra on the sizes (numbers of quantum dots) of arrays is reported and the capacitive coupling between quantum dots in the arrays is explored. PMID- 19546500 TI - Proximity effects in free-standing EBID structures. AB - Proximity effects causing thickening and bending of closely spaced, free-standing pillars grown by electron-beam-induced deposition are investigated. It is shown that growth of a new pillar induces deposition of a layer of additional material on the side of already grown pillars facing the new pillar. We present experimental results which suggest that the bending of pillars is caused by shrinkage of the newly formed layer on exposure to the primary electron beam. PMID- 19546499 TI - Growth of vertical InAs nanowires on heterostructured substrates. AB - We investigate the Au-assisted growth of InAs nanowires on two different kinds of heterostructured substrates: GaAs/AlGaAs structures capped by a 50 nm thick InAs layer grown by molecular beam epitaxy and a 2 microm thick InAs buffer layer on Si(111) obtained by vapor phase epitaxy. Morphological and structural properties of substrates and nanowires are analyzed by atomic force and transmission electron microscopy. Our results indicate a promising direction for the integration of III-V nanostructures on Si-based electronics as well as for the development of novel micromechanical structures incorporating nanowires as their active elements. PMID- 19546501 TI - A plasma sputtering decoration route to producing thickness-tunable ZnO/TiO(2) core/shell nanorod arrays. AB - We report a radio frequency magnetron sputtering method for producing TiO(2) shell coatings directly on the surface of ZnO nanorod arrays. ZnO nanorod arrays were firstly fabricated on transparent conducting oxide substrates by a hydrothermal route, and subsequently decorated with TiO(2) by a plasma sputtering deposition process. The core/shell nanorods have single-crystal ZnO cores and anatase TiO(2) shells. The shells are homogeneously coated onto the whole ZnO nanorods without thickness change. This approach enables us to tailor the thickness of the TiO(2) shell for desired photovoltaic applications on a one nanometer scale. The function of the TiO(2) shell as a blocking layer for increasing charge separation and suppression of the surface recombination was tested in dye-sensitized solar cells. The enhanced photocurrent and open-circuit voltage gave rise to increased photovoltaic efficiency and decreased dark current, indicating successful functioning of the TiO(2) shell. PMID- 19546502 TI - Precise voltage contrast image assisted positioning for in situ electron beam nanolithography for nanodevice fabrication with suspended nanowire structures. AB - In this paper, we demonstrate precise voltage contrast image positioning for in situ electron beam (e-beam) nanolithography to integrate nanowires into suspended structures for nanoswitch fabrication. The positioning of the deflection electrodes on the nanowires can be well controlled using a precise voltage contrast image positioning technique, where the error can be minimized to about 10 nm. Using such a method, dispersed nanowires can be sandwiched between two layers of resist and suspended by one e-beam nanolithography process without any etching. The in situ e-beam nanolithography eliminates the stage movement error by preventing any movements of the stage during the nanolithography process; hence, a high precision laser stage and alignment marks on the substrate are not needed, which simplifies the traditional e-beam nanolithography process. The nanoswitches fabricated using this method show ON and OFF states with the changes of applied voltages. This simplified process provides an easy, low cost and less time-consuming route to integrating suspended nanowire based structures using a converted field emission scanning electron microscope e-beam system, which can also be customized to fabricate multi-layer structures and a site-specific nanodevice fabrication. PMID- 19546503 TI - Three-dimensional selective growth of nanoparticles on a polymer microstructure. AB - We demonstrate a new technique for selectively growing gold nanoparticles on a patterned three-dimensional (3D) polymer microstructure. The technique integrates 3D direct writing of heterogeneous microstructures with nanoparticle synthesis. A digital micromirror device is employed as a dynamic mask in the digital projection photopolymerization process to build the heterogeneous microstructure layer by layer. An amine-bearing polyelectrolyte, branched poly(ethylenimine), is selectively attached to the microstructure and acts as both a reducing and a protective agent in the nanoparticle synthesis. Scanning electron microscopy, energy dispersive x-ray spectroscopy and x-ray photoelectron spectroscopy are utilized to analyze the microstructure and the 3D selectivity of the nanoparticle growth. PMID- 19546504 TI - AFM study of adsorption of protein A on a poly(dimethylsiloxane) surface. AB - In this paper, the morphology and kinetics of adsorption of protein A on a PDMS surface is studied by AFM. The results of effects of pH, protein concentration and contact time of the adsorption reveal that the morphology of adsorbed protein A is significantly affected by pH and adsorbed surface concentration, in which the pH away from the isoelectric point (IEP) of protein A could produce electrical repulsion to change the protein conformation, while the high adsorbed surface protein volume results in molecular networks. Protein A can form an adsorbed protein film on PDMS with a maximum volume of 2.45 x 10(-3) microm(3). This work enhances our fundamental understanding of protein A adsorption on PDMS, a frequently used substrate component in miniaturized immunoassay devices. PMID- 19546505 TI - IL-21 drives secondary autoimmunity in patients with multiple sclerosis, following therapeutic lymphocyte depletion with alemtuzumab (Campath-1H). AB - Phase II clinical trials revealed that the lymphocyte-depleting humanized monoclonal antibody alemtuzumab (Campath-1H) is highly effective in the treatment of early relapsing-remitting multiple sclerosis. However, 30% of patients develop autoimmunity months to years after pulsed exposure to alemtuzumab, usually targeting the thyroid gland and, more rarely, blood components. In this study, we show that autoimmunity arose in those patients with greater T cell apoptosis and cell cycling in response to alemtuzumab-induced lymphocyte depletion, a phenomenon that is driven by higher levels of IL-21. Before treatment, patients who went on to develop secondary autoimmunity had more than 2-fold greater levels of serum IL-21 than the nonautoimmune group. We suggest that serum IL-21 may, therefore, serve as a biomarker for the risk of developing autoimmunity months to years after alemtuzumab treatment. This has implications for counseling those patients with multiple sclerosis who are considering lymphocyte-depleting therapy with alemtuzumab. Finally, we demonstrate through genotyping that IL-21 expression is genetically predetermined. We propose that, by driving cycles of T cell expansion and apoptosis to excess, IL-21 increases the stochastic opportunities for T cells to encounter self antigen and, hence, for autoimmunity. PMID- 19546506 TI - Epididymis seleno-independent glutathione peroxidase 5 maintains sperm DNA integrity in mice. AB - The mammalian epididymis provides sperm with an environment that promotes their maturation and protects them from external stresses. For example, it harbors an array of antioxidants, including non-conventional glutathione peroxidase 5 (GPX5), to protect them from oxidative stress. To explore the role of GPX5 in the epididymis, we generated mice that lack epididymal expression of the enzyme. Histological analyses of Gpx5-/- epididymides and sperm cells revealed no obvious defects. Furthermore, there were no apparent differences in the fertilization rate of sexually mature Gpx5-/- male mice compared with WT male mice. However, a higher incidence of miscarriages and developmental defects were observed when WT female mice were mated with Gpx5-deficient males over 1 year old compared with WT males of the same age. Flow cytometric analysis of spermatozoa recovered from Gpx5-null and WT male mice revealed that sperm DNA compaction was substantially lower in the cauda epididymides of Gpx5-null animals and that they suffered from DNA oxidative attacks. Real-time PCR analysis of enzymatic scavengers expressed in the mouse epididymis indicated that the cauda epididymidis epithelium of Gpx5 null male mice mounted an antioxidant response to cope with an excess of ROS. These observations suggest that GPX5 is a potent antioxidant scavenger in the luminal compartment of the mouse cauda epididymidis that protects spermatozoa from oxidative injuries that could compromise their integrity and, consequently, embryo viability. PMID- 19546507 TI - Dendritic cell entrapment within the pregnant uterus inhibits immune surveillance of the maternal/fetal interface in mice. AB - Embryo implantation induces formation of the decidua, a stromal cell-derived structure that encases the fetus and placenta. Using the mouse as a model organism, we have found that this tissue reaction prevents DCs stationed at the maternal/fetal interface from migrating to the lymphatic vessels of the uterus and thus reaching the draining lymph nodes. Strikingly, decidual DCs remained immobile even after being stimulated with LPS and exhibiting responsiveness to CCL21, the chemokine that drives DC entry into lymphatic vessels. An analysis of maternal T cell reactivity toward a surrogate fetal/placental antigen furthermore revealed that regional T cell responses toward the fetus and placenta were driven by passive antigen transport and thus the tolerogenic mode of antigen presentation that predominates when there is negligible input from tissue resident DCs. Indeed, the lack of involvement of tissue-resident DCs in the T cell response to the fetal allograft starkly contrasts with their prominent role in organ transplant rejection. Our results suggest that DC entrapment within the decidua minimizes immunogenic T cell exposure to fetal/placental antigens and raise the possibility that impaired development or function of the human decidua, which unlike that of the mouse contains lymphatic vessels, might lead to pathological T cell activation during pregnancy. PMID- 19546509 TI - Role of artificial neural networks in dermatology. PMID- 19546508 TI - Mechanical ventilation in patients with end-stage idiopathic pulmonary fibrosis. AB - BACKGROUND: Acute respiratory failure (ARF) occurring during idiopathic pulmonary fibrosis (IPF) is associated with a poor prognosis. In this subset of individuals, mechanical ventilation (MV) may be required. OBJECTIVES: We analysed the characteristics of a group of IPF patients undergoing MV for ARF in order to give some indications on the supposed prognosis. METHODS: Hospital records of 34 consecutive patients with IPF, who underwent MV for ARF, were retrospectively examined. Demographic data, time from diagnosis, gas exchange, Acute Physiology and Chronic Health Evaluation (APACHE) II score, ARF causes and MV failure were recorded. RESULTS: Fifteen subjects (group A) underwent invasive MV and 19 patients (group B) non-invasive ventilation (NIV). The 2 groups were different for disease severity (APACHE II score 24.2 +/- 6 vs. 19.5 +/- 5.9; p = 0.01). Both ventilatory strategies temporarily increased PaO2/FiO2 as compared with spontaneous breathing (group A: 148.5 +/- 52 vs. 99 +/- 39, p = 0.0004; group B: 134 +/- 36 vs. 89 +/- 26, p = 0.0004). NIV reduced the respiratory rate (26 +/- 7 vs. 36 +/- 9 with spontaneous breathing; p = 0.002). Duration of MV correlated with the time of evolution of IPF (r = 0.45; p = 0.018). The in-hospital mortality rate was 85% (100% for invasive MV, 74% for NIV). Four of the 5 survivors died within 6 months from hospital discharge (range 2-6 months). CONCLUSIONS: MV does not appear to have a significant impact on the survival of patients with end-stage IPF. NIV may be useful for compassionate use, providing relief from dyspnoea and avoiding aggressive approaches. PMID- 19546510 TI - Prediction of clinical response to excimer laser treatment in vitiligo by using neural network models. AB - BACKGROUND: A predictive model may help to select likely responders and to anticipate treatment duration in vitiligo. METHODS: We aimed to develop a predictive rule based on data from a randomized trial of excimer laser in vitiligo. Information on 325 treated patches was available. The degree of repigmentation was assessed by digital image analysis of UVB-reflected photographs. Since no strong relationship between any single predictive parameter and outcome was initially documented, we relied on artificial neural networks. RESULTS: Using a time-response optimal threshold model, data were divided into 2 groups of responders and nonresponders. A discriminant network was trained in order to detect responders versus nonresponders. A regression network was subsequently used to compute repigmentation time in responders. The neural network discriminator achieved 66.46 +/- 5.37% (95% CI) overall accuracy. The mean absolute error of the neural network regressor was 19.5843 +/- 2.0930 with a root mean square error of 23.7156 +/- 2.2225. CONCLUSION: Our study offers insight into the difficulty of clinical prediction in vitiligo and presents a way to develop an instrument with which to predict the clinical time response in patients treated by excimer laser. PMID- 19546511 TI - Morphometry, estimation and ultrastructure of ovarian preantral follicle population in queens. AB - The aim of the present study was to estimate the population and morphometrically and ultrastructurally characterize preantral follicles from queen ovaries. Ovaries from 5 queens were collected and processed for light and electron microscopy. A total of 190 preantral follicles (100 primordial, 60 primary and 30 secondary) were analyzed by light microscopy. The diameters of the follicle, oocyte and oocyte nucleus were taken and the number of granulosa cells was counted using a computer program. Queen ovaries presented 37,853 +/- 6,118 preantral follicles on average, with 87% primordial, 10.4% primary and 2.3% secondary follicles. Significant differences were observed in the 3 follicular classes in regard to follicular, oocyte and oocyte nucleus diameters and granulosa cell number (p < 0.05). In regard to ultrastructure, queen preantral follicles presented many unique characteristics, such as early zona pellucida formation in primary follicles and the organization of mitochondria and other organelles in conglomerates and cortical granules aligned at the peripheral zone in secondary follicles. In conclusion, this study described the morphometry and ultrastructure of queen preantral follicles and the preantral follicle population in the ovaries, establishing a pattern for the species and consequently allowing comparisons with other species. PMID- 19546512 TI - Nonirradiated human fibroblasts and irradiated 3T3-J2 murine fibroblasts as a feeder layer for keratinocyte growth and differentiation in vitro on a fibrin substrate. AB - The standard method for producing graftable epithelia relies on the presence of a feeder layer of lethally irradiated 3T3-J2 murine fibroblasts (Rheinwald and Green technique). Here, we studied a new keratinocyte culture system, which envisages the utilization of nonirradiated human fibroblasts embedded into a fibrin substrate, in cultures destined for a future clinical application. We tested this culture system using keratinocytes grown on a fibrin gel precoated with 3T3-J2 murine fibroblasts as a control. In order to evaluate the new technology, we compared the clonogenic potential and the proliferative, differentiative and metabolic characteristics of keratinocytes cultured on the fibrin gel under the two culture conditions. The results demonstrated that the proposed technology did not impair the behavior of cultured keratinocytes and revealed that cells maintained their proliferative potential and phenotype under the experimental conditions. In particular, the demonstration of stem cell maintenance under the adopted culture conditions is very important for acute burn treatment with skin substitutes. This work is a first step in the evaluation of a new keratinocyte culture system, which has been studied in order to take advantage of an additional human cell population (i.e. nonirradiated, growing fibroblasts) for future transplantation purposes in acute and chronic wounds. Additional research will allow us to attain (1) the removal of murine cells in the initial phase of keratinocyte cultures, and (2) the removal of other potentially dangerous animal-derived materials from the entire culture system. PMID- 19546513 TI - The shock of aging: molecular chaperones and the heat shock response in longevity and aging--a mini-review. AB - BACKGROUND: Aging can be thought of as the collision between destructive processes that act on cells and organs over the lifetime and the responses that promote homeostasis, vitality and longevity. However, the precise mechanisms that determine the rates of aging in organisms are not known. OBJECTIVE: Macromolecules such as proteins are continuously exposed to potential damaging agents that can cause loss of molecular function and depletion of cell populations over the lifetime of essential organs. One of the key homeostatic responses involved in maintaining longevity is the induction of heat shock proteins (HSPs), a conserved reaction to damaged intracellular proteins. We aim to discuss how the interplay between protein damage and its repair or removal from the cell may influence longevity and aging. METHODS: We have reviewed experiments carried out in mammalian and non-mammalian organisms on molecular chaperones and the transcription factor (heat shock factor 1, HSF1) responsible for their expression. We have discussed mechanisms through which these molecules are regulated in cells, respond to stimuli that enhance longevity and become impaired during aging. RESULTS: The transcription factor HSF1 initiates the prolific induction of HSP when cells are exposed to protein damage. HSPs are molecular chaperones that protect the proteome by folding denatured polypeptides and promoting the degradation of severely damaged proteins. Activation of HSF1 is coupled functionally to fundamental pathways of longevity and orchestrates the evasion of aging through HSP induction and antagonism of protein aggregation. In addition to mediating protein quality control, some HSPs such as Hsp27 and Hsp70 directly protect cells against damage-induced entry into death pathways. However, the heat shock response declines in potency over the lifetime, and enfeeblement of the response contributes to aging by permitting the emergence of protein aggregation diseases, reduction in cellular vigor and decreased longevity. CONCLUSIONS: Molecular chaperones play an important role in the deterrence of protein damage during aging and their expression is required for longevity. Chemical stimulation of HSP synthesis might therefore be a significant strategy in future design of antiaging pharmaceuticals. PMID- 19546514 TI - Comparative proteome analysis of splenic lymphocytes in senescence-accelerated mice. AB - BACKGROUND: Senescence-accelerated mice (SAM) are commonly used as animal models for studying aging. To date, aging-related proteomes in the livers and brains of SAMP8 mice have been reported; however, spleen of SAM has not yet been used as material for studying the aging-related proteome despite its extremely important role in the progress of aging or aging-related diseases. OBJECTIVE: To identify proteins associated with aging or aging-related diseases in SAM. METHODS: Two dimensional gel electrophoresis (2DE) was used to compare the proteomes of splenic lymphocytes from two strains of SAM (SAMP8 and SAMR1). Differentially expressed spots whose expression altered over twofold (p < 0.01) were identified by LC-MS/MS and database search. RESULTS: Identification results showed that 18 differentially expressed protein spots represented 14 different proteins known to be involved in aging- or aging disease-related pathways, i.e., annexin I, calgranulin B, transaldolase, isocitrate dehydrogenase, and an unnamed protein product were involved in oxidative stress and cell defense, phosphoglycerate mutase I, aldose reductase, carbonic anhydrase I and carbonic anhydrase II were involved in glycolytic energy metabolism and homeostasis, and macrophage capping protein and high mobility group box-1 protein were involved in infection and DNA damage. CONCLUSION: 2DE combined with MS is effective for comparative proteome analysis of splenic lymphocytes of SAM to identify proteins closely involved in aging or aging-related diseases, especially immune diseases, and further functional studies of these proteins in SAM may throw some light on the study of biomarkers for aging or aging-related diseases. PMID- 19546515 TI - Art and aging. AB - The study explores the process of aging in artists as reflected in their creative output after the age of 60. Additionally, a wide range of artists who died young are also included, as their late work, i.e. the work of their last years of life, is very similar to the late work of their older colleagues. Therefore, it is quite right to describe it as work done later in life (in relation to their age). PMID- 19546516 TI - Characterization of human mesenchymal stem cell-engineered cartilage: analysis of its ultrastructure, cell density and chondrocyte phenotype compared to native adult and fetal cartilage. AB - The production of engineered cartilage from mesenchymal stem cells is a rapidly developing field. Potential applications include the treatment of degenerative joint disease as well as the treatment of traumatic and surgical bone injury. Prior to clinical application, however, further characterization of the morphology, ultrastructure, biocompatibility, and performance of the engineered tissue is warranted. To achieve this, human mesenchymal stem cells (hMSCs) were grown in vitro in pellet culture for 3 weeks in chondrogenic medium conditions. The resultant engineered cartilage was compared to native adult and fetal tissue. Routine histology, special stains, and ultrastructural and quantitative histomorphometric analyses were performed. The engineered tissue demonstrated a similar chondrocyte phenotype, collagen fibril appearance, and matrix distribution when compared to native cartilage. By histomorphometric analysis, the cell density of the engineered cartilage was between that of native fetal and adult cartilage. The cell-to-matrix ratio and cellular area fraction of engineered cartilage samples was significantly greater than in adult samples, but indistinguishable from fetal cartilage samples, supporting the hypothesis that hMSC-engineered cartilage regeneration may mimic fetal cartilage development. PMID- 19546517 TI - Effects of chronic intracerebral prolactin on the oxytocinergic and vasopressinergic system of virgin ovariectomized rats. AB - Chronic intracerebroventricular (icv) infusion of prolactin (PRL) into the cerebral ventricles and mimicking central hyperprolactinemia in lactation has recently been shown to reduce anxiety and neuronal as well as neuroendocrine responses to acute stressor exposure. Here, we studied the effects of icv PRL on the activity of the oxytocin (OXT) and arginine vasopressin (AVP) systems of virgin female, ovariectomized, estradiol-substituted Wistar rats. Ovine PRL was delivered via osmotic minipumps at 0.01, 0.1 or 1 microg/h for 5 days. Under basal conditions, both plasma OXT and AVP concentrations were increased after chronic PRL treatment (1 microg/h). At hypothalamic level, this was accompanied by an increased c-fos and OXT mRNA expression within the supraoptic nucleus, the main source of plasma OXT, whereas AVP mRNA levels remained unchanged. No effect of PRL on c-fos or on nonapeptide mRNA expression was found in the hypothalamic paraventricular nucleus. Moreover, chronic PRL abolished the rise in plasma OXT induced by acute exposure to 30 min restraint stress in vehicle-treated rats. However, restraint stress did not significantly alter OXT or AVP mRNA expression in the hypothalamus of either vehicle- or PRL-treated animals. From these results we conclude that brain hyperprolactinemia alters the synthetic activity of OXT neurons and the secretory performance of OXT and AVP neurons within the hypothalamus, resulting in elevated plasma concentrations of both hormones under basal conditions. These changes are comparable to adaptations seen in the female peripartum period. PMID- 19546518 TI - Radiological versus histopathological diagnosis of usual interstitial pneumonia in the clinical practice: does it have any survival difference? AB - BACKGROUND: Recent studies have shown that quantification of specific histopathologic features found in usual interstitial pneumonia (UIP) are useful in defining a prognosis, suggesting the need of biopsy in all patients. OBJECTIVES: This study examines whether UIP-associated mortality is different in patients diagnosed by high-resolution computed tomography (HRCT) features considered definite of UIP and in patients with no definite radiological diagnosis that required histological confirmation of diagnoses. METHODS: Forty five patients were included (30 males, mean age 65.3 +/- 10.7 years). Two groups of patients were identified: those with HRCT findings of definite UIP (n = 26) and those whose radiological diagnosis was not definite and required a surgical biopsy to confirm the presence of UIP (n = 19). Forced vital capacity, forced expiratory volume in 1 s and diffusing capacity for carbon monoxide were measured in all patients. All data were obtained from medical records, and the survival status was obtained by telephone or personal interview. All clinical parameters and HRCTs were obtained within 1 month before surgical lung biopsy. RESULTS: Median survival was not different across groups and was similar to that previously reported (35 months). Kaplan-Meier analysis did not show any difference in 5-year survival between both groups. CONCLUSIONS: In a clinical context in which the diagnosis of UIP can be obtained as a dynamic process that includes an integrated clinical, radiological and pathologic approach, a reliable diagnosis of UIP can be obtained based on a typical definite HRCT with no risk of including patients with a more benign disease and a more prolonged survival. PMID- 19546519 TI - Accuracy and feasibility of electromagnetic navigated bronchoscopy under nitrous oxide sedation for pulmonary peripheral opacities: an outpatient study. AB - BACKGROUND: Recent studies have described the promising method of electromagnetic navigated bronchoscopy (ENB) for diagnosis of peripheral solitary nodules. However, they require general anaesthesia or intravenous sedation. We wanted to know if ENB could be applied more easily in outpatients. OBJECTIVES: We prospectively evaluated the accuracy and the feasibility of ENB under local anaesthesia and nitrous oxide/oxygen inhalation as unique sedation in outpatients. METHODS: After mapping time, the bronchoscopic procedure was carried out under local anaesthesia and nitrous oxide/oxygen inhalation with the unique help of the ENB to confirm the right position of the extended working channel before sampling. The primary end point was the accuracy of ENB and the secondary end point was the feasibility in outpatients. RESULTS: Among 54 screened patients, 53 completed the study protocol. The overall diagnostic success rate to diagnose malignancy was 71.4% in tumours of 28 mm in median size. ENB classified correctly peripheral lesions according to malignity in 41 cases (30 cases of cancer, 11 benign diagnosis) and failed in 12 cases (1 probable lung metastasis, 11 lung cancers). All patients but 1 were dismissed 1 h after the procedure and the tolerance of the procedure was excellent in all cases except 2 (agitation and anxiety). In two cases (4%) a pneumothorax was recorded, 1 requiring drainage with a chest tube during a short hospitalisation. CONCLUSIONS: ENB under nitrous oxide/oxygen sedation seems to be an accurate and safe procedure. In our series, it allowed us to obtain the diagnosis in 71.4% of the tumours, with a good tolerance and an outpatient strategy. PMID- 19546520 TI - Genuair is a good inhaler but an indirect comparison with other dry powder inhalers is inadequate. PMID- 19546521 TI - Small cell carcinoma with paraneoplastic polyneuropathy and tumor embolization: a case report and literature review. AB - Although small cell cancer of the lung may have protean manifestations, tumor embolization and inability to identify the primary sites are unusual features. We present a patient with anti-Hu antibody-associated paraneoplastic sensory polyneuropathy and tumor embolism diagnosed by endovascular biopsy to be due to small cell cancer, the primary site of which was not evident. To our knowledge, this patient represents only the second individual to be described with tumor embolization complicating small cell cancer and reminds clinicians of the extended spectrum of this disease. PMID- 19546522 TI - A comparison of mycophenolate mofetil with ciclosporine for the treatment of chronic plaque-type psoriasis. AB - AIMS: To compare the efficacy of ciclosporine (CsA) versus mycophenolate mofetil (MMF) in psoriasis, a randomized trial was conducted. METHODS: A prospective multicenter randomized open-label clinical trial was performed to compare two parallel groups of patients with chronic plaque psoriasis undergoing different treatments. Therefore, a total of 54 patients with psoriasis were randomly assigned to treatment with either CsA (2.5 mg/kg body weight) or MMF (2 g daily) for 12 weeks, and the drug doses were adjusted according to response. The psoriasis area and severity index (PASI) was used to assess the clinical severity of psoriasis. The primary outcome of this trial was the time to disease relapse. Safety, PASI scores and psoriasis disability index (PDI) were assessed as secondary outcome. RESULTS: There was no difference in time to disease relapse between the two groups. After 12 weeks of treatment, the mean PASI score (+/-SD) decreased from 24.6 +/- 11.1 to 6.6 +/- 7.3 in the CsA group (n = 27) and from 22.4 +/- 9.2 to 10.6 +/- 6.7 in the MMF group (n = 27; p = 0.02). The side effects, time to remission and PDI were similar in both groups. CONCLUSIONS: After 12 weeks, CsA demonstrated a significantly superior efficacy in psoriasis compared to MMF. PMID- 19546523 TI - Granulomatous slack skin responds to UVA1 phototherapy. AB - Granulomatous slack skin (GSS) is an extremely rare disorder within the group of cutaneous T cell lymphomas (CTCL). Ultraviolet A1 (UVA1) phototherapy has previously been reported to be useful in the treatment of CTCL such as mycosis fungoides. We report a 35-year-old Caucasian male with GSS treated with UVA1 phototherapy starting at 20 J/cm(2) UVA1 3 times a week and subsequently increased in increments of 5 J/cm(2) to a medium-range dose of 50 J/cm(2) per session. The patient underwent a total of 45 sessions with a cumulative dose of 1,495 J/cm(2) UVA1 without any adverse events. At the conclusion of UVA1 phototherapy, a decrease in erythema and skin thickness was observed which was most prominent in the periphery of the lesion in the right groin area. A follow up 12 months after phototherapy showed continued treatment benefit. To our knowledge, this is the first report describing the successful use of UVA1 (340 400 nm) phototherapy in a patient with GSS. PMID- 19546524 TI - Prophylactic use of granulocyte colony-stimulating factor after chemotherapy does not affect survival rate in acute myeloid leukemia: a meta-analysis. PMID- 19546525 TI - Accurate prenatal diagnosis of Hb Bart's hydrops fetalis in daily practice with a double-check PCR system. AB - Hemoglobin (Hb) Bart's hydrops fetalis is a fatal condition associated with homozygous alpha(0)-thalassemia. Prenatal diagnosis of the disease is usually done by gap-PCR; however, misdiagnosis can occur with allelic dropout. Diagnosis using more than one method is preferred. We describe a double-check PCR assay for accurate prenatal diagnosis. The study was conducted on 64 fetuses at risk of homozygous alpha(0)-thalassemia encountered at our routine thalassemia diagnosis laboratory. Chorionic villus sample (CVS), amniotic fluid or fetal blood specimens were obtained from pregnant women at risk and analyzed by two PCR methods. In the first method, the SEA alpha(0)-thalassemia deletion of parents and fetuses were determined by gap-PCR routinely run in our laboratory. In another method, two specific fragments located 5' to the zeta(2) gene (XbaI fragment) and the alpha(2)-globin gene (RsaI fragment) together with the gap-PCR fragment were multiply co-amplified to determine the presence or absence of normal and alpha(0)-thalassemia alleles. The molecular diagnosis of alpha(0) thalassemia was possible in all 64 fetuses using the two PCR approaches. The final diagnoses included 13 normal, 29 unaffected heterozygote and 22 homozygote alpha(0)-thalassemia fetuses.The two PCR assays disclosed no discordant result in the diagnosis of the Hb Bart's hydrops fetalis caused by alpha(0)-thalassemia.The combined PCR assay for gap-PCR, zeta(2) XbaI and alpha(2) RsaI fragments, described here, is simple, accurate and applicable in the prenatal diagnosis of Hb Bart's hydrops fetalis in a routine setting. PMID- 19546526 TI - Ischemic preconditioning attenuates renal ischemia-reperfusion injury by inhibiting activation of IKKbeta and inflammatory response. AB - BACKGROUND: Renal ischemia-reperfusion (I/R) injury is a major cause of acute renal failure (ARF). The transcription factor nuclear factor-kappaB (NF-kappaB) has been implicated as a key mediator of reperfusion injury. Activation of NF kappaB is dependent upon the phosphorylation of its inhibitor, IkappaB, by the specific inhibitory kappaB kinase (IKK) subunit, IKKbeta. We hypothesized that ischemic preconditioning (IPC) reduces acute renal damage following I/R injury by inhibiting activation of IKKbeta. As neutrophil gelatinase-associated lipocalin (NGAL), an early predictive biomarker of acute kidney injury, is regulated by NF kappaB, we approached the relationship between NGAL and IKKbeta. METHOD: Thirty male Sprague-Dawley rats were randomly divided into 3 groups after right kidney nephrectomy. Group A rats were sham-operated controls. Group B rats were 45-min ischemic in the left renal artery while Group C rats were pre-treated with 3 cycles of 2-min ischemia and 5-min reperfusion. All the rats were sacrificed at 24 h after reperfusion. We harvested kidneys and serum to do further analysis, including histological and functional parameters, expressions of NGAL and IKKbeta in renal tissues. RESULTS: Compared with rats subjected to I/R injury, pre treated rats had a significant decrease in serum creatinine level (Scr) and tubulointerstitial injury scores (Scr, 86.79 +/- 12.98 vs. 205.89 +/- 19.16 mircomol/l, p < 0.01; tubulointerstitial injury scores, 1.3 +/- 0.48 vs. 3.8 +/- 0.79, p < 0.01). In addition, expressions of IKKbeta (0.95 +/- 0.21 vs. 1.74 +/- 0.17, p < 0.05) and NGAL (1.71 +/- 0.032 vs. 2.66 +/- 0.078, p < 0.05) at renal tubule in pre-treated rats were attenuated significantly compared with rats subjected to ischemia-reperfusion injury. Moreover, our study showed that IKKbeta and NGAL were in positive correlation (R = 0.965 > R(0.01)(30) = 0.448, p < 0.01). CONCLUSIONS: The evidence suggests that IKKbeta may play a role in renal I/R injury and give rise to the generation of NGAL. It appears that IPC may attenuate renal injury and the expression of NGAL following acute I/R injury. IKKbeta may offer a clinically accessible target for preventing renal injury following I/R. PMID- 19546527 TI - Effects of new peritoneal dialysis solutions, pyridoxamine and AT1 receptor blocker, on TGF-beta1 and VEGF expression in rat peritoneal mesothelial cells. AB - BACKGROUND: Transforming growth factor beta1 (TGF-beta1) and vascular endothelial growth factor (VEGF) are involved in peritoneal deterioration in continuous ambulatory peritoneal dialysis. The present study was designed to determine whether new peritoneal dialysis solutions (PDS), pyridoxamine (advanced glycation end products (AGE) inhibitor) or AT1 receptor blocker (ARB), affect the expression of VEGF and TGF-beta1 in rat peritoneal mesothelial cells (RPMC). METHODS: RPMC were stimulated by phosphate-buffered saline (PBS) as control, Dianeal 1.5% (D 1.5%), Dianeal 2.5% (D 2.5%), Dianeal 4.25% (D 4.25%), Dianeal N 1.5% (N 1.5%), Dianeal N 2.5% (N 2.5%) or Extraneal (Ex). In co-incubation experiments, RPMC were stimulated with N 2.5% including pyridoxamine or olmesartan (ARB). VEGF and TGF-beta1 protein and mRNA expression in RPMC were analyzed by ELISA and RT-PCR. RESULTS: Glucose-containing PDS, even N 2.5% diluted twofold with M199 (which contains 1.25% glucose), increased VEGF and TGF beta1 expression in RPMC (p < 0.05). Ex did not inhibit VEGF expression and did not inhibit TGF- beta1 expression after 24 h in RPMC. Pyridoxamine and ARB significantly reduced N 2.5%-induced VEGF and TGF-beta1 protein and mRNA expression in RPMC (p < 0.01). CONCLUSIONS: Neither new pH-neutral, lactate buffered, low-GDP, two-chamber bag PDS, nor 7.5% icodextrin PDS alone satisfactorily inhibited VEGF and TGF-beta1 overproduction in RPMC, but ARB or pyridoxamine effectively inhibited glucose-containing PDS (N 2.5%)-induced overproduction. PMID- 19546529 TI - Lack of association between small dense low-density lipoprotein levels and coronary artery disease in chronic hemodialysis patients. AB - BACKGROUND/AIMS: Small, dense low-density lipoprotein (sd-LDL) is a newly recognized risk factor for coronary artery disease (CAD) in the general population. This study was performed to evaluate the relationship between sd-LDL, and CAD and cardiovascular risk factors such as other lipid profiles and hemostatic factors, in patients with chronic hemodialysis (CHD). METHODS: Thallium single-photon emission computed tomography (SPECT) was performed to evaluate for CAD in 126 CHD patients. Coronary angiography was performed in patients with positive thallium SPECT. CHD patients were classified into CAD and non-CAD group. LDL subfractions, other lipid profiles, and hemostatic factors were measured. RESULTS: The proportion of sd-LDL, and mean LDL size did not differ between CHD patients and healthy controls. Twenty-eight CHD patients had CAD by thallium SPECT and coronary angiography. The proportion of sd-LDL and mean LDL size did not differ between CAD and non-CAD patients. Age, diabetes mellitus presentation and high-sensitivity C-reactive protein (hs-CRP) levels were significantly higher, and prealbumin and apolipoprotein A1 levels were significantly lower, in the CAD group (p < 0.05). The proportion of sd-LDL was positively correlated with triglyceride levels (p < 0.001), apolipoprotein B (p < 0.05) and fibrinogen (p < 0.05). CONCLUSION: This study showed that sd-LDL is not increased in CHD patients and is not associated with CAD in such patients. PMID- 19546528 TI - A synergistic association of ACE I/D and eNOS G894T gene variants with the progression of immunoglobulin A nephropathy - a pilot study. AB - BACKGROUND: Individual variability in the natural history and response to therapy of immunoglobulin A nephropathy (IgAN) suggests a complex multifactorial pathogenesis. We investigated whether single nucleotide polymorphisms (SNPs) involved in the non-immunologic progression of renal disease are related with disease progression. METHODS: This is a pilot historic cohort study of 64 Caucasian patients with biopsy-proven IgAN and a median follow-up of 70 months. Three SNPs of the renin-angiotensin system genes (angiotensin I converting enzyme insertion/deletion (ACE I/D), angiotensinogen (AGT) M235T, and angiotensin II type 1 receptor (AT(1)R) 1166A/C), 2 of the endothelial nitric oxide synthase (eNOS), 4a/b and G894T, and 1 of the bradykinin 1 receptor, G-699C, were genotyped. The primary outcome was 'kidney survival' defined as a 30% decrease of baseline creatinine clearance; annualized decrease of glomerular filtration rate was also calculated. RESULTS: Proteinuria, histological lesions, and mean arterial pressure were related to an unfavorable outcome. The simultaneous presence of the DD and GG variants of the ACE and eNOS genes was related to an unfavorable outcome as compared with other combinations [hazard ratio ranging from 4.7 (95% CI 1.52-14.33) to 8.4 (95% CI 2.45-29.10)] after controlling for proteinuria, mean arterial pressure and baseline histological lesions. CONCLUSION: This study suggests that in our population with IgAN, an interaction between ACE and eNOS polymorphisms may be a prognostic factor for renal function deterioration. PMID- 19546530 TI - Efficacy and safety of glucocorticoids therapy for IgA nephropathy: a meta analysis of randomized controlled trials. AB - BACKGROUND: Published reports examining the efficacy and safety of glucocorticoids for preserving renal function in immunoglobulin A nephropathy (IgAN) have yielded conflicting results. To systematically evaluate the efficacy and safety of glucocorticoids agents for IgAN, we conducted a meta-analysis of the published randomized controlled trials (RCTs). METHODS: MEDLINE, EMBASE, the Cochrane Library and article reference lists were searched for RCTs that compared glucocorticoids with placebo and any other non-immunosuppressive agents for treating IgAN. The quality of the studies was evaluated with the method of the intention-to-treat analysis and allocation concealment, as well as with the Jadad method. Meta-analyses were performed on the outcomes of proteinuria and renal survival and adverse events in patients with IgAN. RESULTS: Seven RCTs involving 386 patients were included in the review. Four RCTs reported renal survival data with Kaplan-Meier survival curves. Overall, glucocorticoid agents had statistically significant effects on improved renal survival (HR 0.20, 95% CI 0.20 to 0.39) and reduction of proteinuria (standardized mean difference SMD, 0.51; 95% CI -0.73 to -0.29) when compared with the control group. Tests for heterogeneity showed no difference in effect among the studies. In general, glucocorticoid agents were well tolerated. Patients receiving glucocorticoids therapy did not have an increased risk of development of type 2 diabetes mellitus, hypertension or Cushingoid adverse effects, while glucocorticoids were associated with a significant increase in the risk of gastrointestinal tract adverse events. CONCLUSIONS: The current cumulative evidence suggests that glucocorticoids have statistically significant effects on protecting renal function and reduction of proteinuria in patients with IgAN, but we should be careful for its gastrointestinal tract reaction. In general, glucocorticoids agents are a promising medication and should be investigated further. PMID- 19546532 TI - The forebrain of actinopterygians revisited. AB - The forebrain of actinopterygian fishes differs from that of other vertebrates in that it consists of a pair of solid lobes. Lateral ventricles surrounded by nervous tissue are entirely lacking. Comparative anatomical and embryological studies have shown that the unusual configuration of the forebrain in actinopterygians results from an outward bending or eversion of the dorsal portions of its lateral walls. Due to this eversion, the telencephalic roof plate is transformed into a wide, membranous structure which surrounds the dorsal and lateral parts of the solid lobes and is attached to their lateral or ventrolateral aspects. The taeniae, i.e. the lines of attachment of the widened roof plate, represent important landmarks in actinopterygian forebrains. In the present paper, the process of eversion is specified and quantified. It is pointed out that recent suggestions to modify the original eversion concept lack an empirical basis. Eversion is the antithesis of the inward bending or inversion that occurs in the forebrains of most other vertebrates. The forebrain lobes in actinopterygians, like those in other vertebrates, comprise a pallium and a subpallium, both of which include a number of distinct cell masses. The morphological interpretations of these cell masses over the past 130 years are reviewed and evaluated in light of a set of carefully selected criteria for homologous relationships. Special attention is paid to the interpretation of a cell mass known as Dp, situated in the caudolateral portion of the pallium in teleosts (by far the largest clade of living actinopterygians). Based on its position close to the taenia, and given the everted condition of the pallium in teleosts, this cell mass clearly corresponds with the medial pallium in inverted forebrains; however, Dp receives a dense olfactory input, and it shares this salient feature with the lateral pallium, rather than with the medial pallium of inverted forebrains. There is presently no consensus regarding the homology of Dp. Several recent authors [Wullimann and Mueller, 2004; Yamamoto et al., 2007] consider the lateral pallium in inverted forebrains and Dp in teleosts to be homologous because they believe that these cell masses originate from the same germinative zones, but that Dp attains its ultimate position only through migration. On the other hand, the present author believes that Dp is situated in the immediate vicinity of its germinative zone and that it represents a specialized part of the lateral pallial zone in teleosts, a zone that can be homologized topologically with the medial pallium in inverted forebrains. Further, it is proposed that the lateral olfactory tract in teleosts, which supplies most of the olfactory fibers to Dp, is not homologous to the same-named tract in the inverted forebrains of most other vertebrates. PMID- 19546531 TI - An architectonic study of the neocortex of the short-tailed opossum (Monodelphis domestica). AB - Short-tailed opossums (Monodelphis domestica) belong to the branch of marsupial mammals that diverged from eutherian mammals approximately 180 million years ago. They are small in size, lack a marsupial pouch, and may have retained more morphological characteristics of early marsupial neocortex than most other marsupials. In the present study, we used several different histochemical and immunochemical procedures to reveal the architectonic characteristics of cortical areas in short-tailed opossums. Subdivisions of cortex were identified in brain sections cut in the coronal, sagittal, horizontal or tangential planes and processed for a calcium-binding protein, parvalbumin (PV), neurofilament protein epitopes recognized by SMI-32, the vesicle glutamate transporter 2 (VGluT2), myelin, cytochrome oxidase (CO), and Nissl substance. These different procedures revealed similar boundaries among areas, suggesting that functionally relevant borders were detected. The results allowed a fuller description and more precise demarcation of previously identified sensory areas, and the delineation of additional subdivisions of cortex. Area 17 (V1) was especially prominent, with a densely populated layer 4, high myelination levels, and dark staining of PV and VGluT2 immunopositive terminations. These architectonic features were present, albeit less pronounced, in somatosensory and auditory cortex. The major findings support the conclusion that short-tailed opossums have fewer cortical areas and their neocortex is less distinctly laminated than most other mammals. PMID- 19546534 TI - Telestroke: the use of telemedicine in stroke care. Preface. PMID- 19546533 TI - Biases in measuring the brain: the trouble with the telencephalon. AB - When correlating behavior with particular brain regions thought responsible for the behavior, a different region of the brain is usually measured as a control region. This technique is often used to relate spatial processes with the hippocampus, while concomitantly controlling for overall brain changes by measuring the remainder of the telencephalon. We have identified two methods in the literature (the HOM and TTM) that estimate the volume of the telencephalon, although the majority of studies are ambiguous regarding the method employed in measuring the telencephalon. Of these two methods, the HOM might produce an artificial correlation between the telencephalon and the hippocampus, and this bias could result in a significant overestimation of the relative hippocampal volume and a significant underestimation of the telencephalon volume, both of which are regularly used in large comparative analyses. We suggest that future studies should avoid this method and all studies should explicitly delineate the procedures used when estimating brain volumes. PMID- 19546535 TI - Telemedicine in stroke: organizing a network--rationale and baseline principles. AB - Telestroke is the specific term used for the application of telemedicine in stroke. It is a consultative modality that facilitates care of patients with acute stroke at underserved hospitals by specialists at stroke centers and can play a vital role in minimizing the overall medicosocial impact of stroke. Telestroke should not be viewed as a new form of stroke therapy, rather as a means of supporting the increased delivery of evidenced-based medicine in stroke. The design and implementation of a hub-and-spoke telestroke network are complex and require state-of-the-art technology and close, organized collaboration between healthcare professionals if they are to be achieved. Telestroke is becoming part of clinical practice in some regions. It provides rapid access to specialized treatment and it could also potentially lead to major improvements in basic on-site management. Telemedicine is also being used for secondary prevention, rehabilitation, education and long-term stroke care. However, for progress to continue and in order to ensure the long-term sustainability of telestroke, various medicolegal, economic and market issues need to be resolved. PMID- 19546536 TI - Use of telemedicine to increase thrombolysis and advance care in acute ischemic stroke. AB - The use of the only proven therapy for acute ischemic stroke, intravenous tissue plasminogen activator (tPA), remains disappointingly low. One potential way to increase the use of tPA is by the implementation of telemedicine stroke care networks. Preliminary data from several studies indicate that the safe and expanded use of tPA for ischemic stroke can be accomplished with the help of telemedicine. Telemedicine stroke care networks can also be used in the future to enhance stroke diagnosis with advanced CT and MRI technology and to potentially increase the number of patients referred to tertiary stroke centers for intra arterial therapies. It is highly likely that telemedicine stroke care will substantially enhance acute stroke therapy for remote and underserved populations. PMID- 19546537 TI - Telestroke: scientific results. AB - Telestroke is the application of telemedicine in stroke care. Hence, teleconsultation means the performance of consultation by a remotely located expert through the use of high-quality videoconferencing. Remote evaluation of stroke patients via telemedicine is increasingly utilized, particularly in neurologically underserved areas. It is usually based on video examination and evaluations of brain scans via teleradiology. Scientific analyses have demonstrated the reliability of neurological assessments via videoconference. Teleradiology using electronically transmitted original imaging data is potentially equivalent to onsite assessment. Telemedicine consultation including video examination and teleradiology improves accuracy of acute stroke treatment decisions compared to telephone-based consultation. Telemedicine is already frequently used to extend the benefits of intravenous thrombolysis to patients in nonspecialized hospitals. Beyond thrombolysis, telemedicine can be used for immediate decision making in acute stroke, e.g. for triage of patients who might benefit from interventional treatments not available at the referring hospital. However, improved clinical outcomes of stroke patients have only been investigated and shown when telemedicine was combined with the Stroke Unit concept based on specialized stroke wards and organized stroke care. More scientific evaluation is needed in the fields of cost effectiveness, quality management and implementation of further technological innovations. There are still insufficient data about the use of telemedicine in stroke prevention, rehabilitation and post-stroke care. PMID- 19546538 TI - Telestroke networking offers multiple benefits beyond thrombolysis. AB - Patients with acute ischemic stroke (AIS) require immediate attention and stroke expertise, which is rarely offered by community hospitals. Telestroke networks were originally established for delivering thrombolysis to inhabitants of underserved regions where stroke expertise was not available 24/7. Rapidly expanding experience addresses the fact that thrombolysis, when given using telestroke consultation, is as safe and effective as when it is given in a stroke center. Telestroke, without a doubt, increases the number of patients receiving thrombolysis, and thus improves patient outcomes, but additionally these networks together with a comprehensive organization of acute stroke care could bring many other benefits which so far are heavily underused in neurology. These benefits include: shortening hospital stay of patients through advanced care, avoiding a large number of unnecessary patient transfers, identifying specific stroke patients who require urgent interventions or surgery (such as subarachnoid hemorrhage, intraventricular hemorrhage, candidates for craniectomy, or basilar artery occlusion), leading to establishment of stroke units and stroke teams in spoke hospitals and overall improvement of stroke care in spoke hospitals, early diagnosis and proper treatment of stroke and nonstroke patients. Further benefits may be: to facilitate staff recruitment to spoke hospitals, to deliver expertise to developing countries, participation of spoke hospitals to acute stroke treatment trials and stroke prevention trials, and environmental effects. The magnitude of these benefits will become more obvious in the near future because this exciting field is progressing fast. The Finnish experience suggests that telestroke is a versatile tool for improving acute stroke care of inhabitants in underserved regions and it should be made more widely available. PMID- 19546539 TI - Telestroke in stroke survivors. AB - Stroke is a high-frequency disorder placing a significant burden on the health care systems, being the foremost cause of complex chronic disability in adults. Devising systems that can enhance the prevention of stroke recurrence is an important priority and challenge in both the developed and the developing world. The potential for recurrent stroke can be substantially reduced by effective management of vascular risk factors. Telestroke is a tool with potential application to improve risk management of stroke survivors. Lack of acknowledgment of existing practices as well as lack of awareness of potential financial barriers to diffusion of telestroke can lead to limited implementation. Telestroke offers service providers the opportunity to access large numbers of stroke survivors targeting secondary prevention. The ideal 'telestroke model' provides service support, education for the patient and caregiver, as well as integration of specialist and primary care services. Effective use of technological advances, with adequate recognition of the importance of human interaction in the long-term management of a largely elderly population of stroke survivors is challenging but possible. Telestroke should be systems- and not technology-driven. Barriers in the implementation of telestroke have been identified as insufficient planning of IT infrastructure, lack of long-term vision for sustainability, a lack of contextual perspective as well as poor communication across domains. Future telestroke models should provide effective action in an integrated model of care recognizing and involving all existing players and practices. PMID- 19546540 TI - Barriers, legal issues, limitations and ongoing questions in telemedicine applied to stroke. AB - The use of telemedicine services, such as telestroke, is still highly fragmented and its deployment in an integrative healthcare system is challenging. Factors impeding the growth of telemedicine include confidence and malpractice issues, technical advances, reimbursement, licensing, credentialing costs, cost effectiveness, and legal issues. These barriers, limitations and requirements in the routine use of telemedicine are reviewed, in addition to medical activities, the objectives of telestroke, technical aspects, funding, legal issues, evaluation and quality management. As telemedicine induces a new form of interrelationship between health care providers, mutual trust and acceptance need to be developed in telemedicine services. Furthermore, education and training will be crucial in order to facilitate the use of telestroke over the next decade. PMID- 19546541 TI - Endogenous activated protein C predicts hemorrhagic transformation and mortality after tissue plasminogen activator treatment in stroke patients. AB - BACKGROUND: Activated protein C (APC) is a plasma serine protease with systemic anticoagulant and a wide spectrum of cytoprotective activities that has been proposed as a promising therapy for acute stroke. Therefore, we sought to investigate the role of endogenous APC in human ischemic stroke. METHODS: Our target were 119 consecutive patients with an ischemic stroke involving the middle cerebral artery territory who received tissue plasminogen activator (t-PA) within 3 h of symptom onset. APC was measured before, as well as 1 and 2 h after t-PA administration, and again at 12 and 24 h after stroke onset. Cranial tomography scan was obtained at admission and repeated at 24-48 h or when a neurological worsening occurred to rule out the presence of hemorrhagic complications. The functional outcome was evaluated by 3-month modified Rankin Scale. RESULTS: A total of 117 t-PA-treated patients were finally included in the analyses. APC peaked at 1 h after t-PA administration (pretreatment APC = 132.44 +/- 36.39%, 1 hour APC = 184.20 +/- 34.28%, 2-hour APC = 145.50 +/- 35.23%; p < 0.0001). Interestingly, a high 2-hour APC level was associated with parenchymal hemorrhages (OR = 25.19; 95% CI = 4.76-133.19; p = 0.0001) and mortality (OR = 13.8; 95% CI = 2.58-73.63; p = 0.001), in a logistic regression model. Our results remained significant after Bonferroni correction for multiple testing. CONCLUSIONS: A high endogenous APC level 2 h after t-PA administration is independently associated with hemorrhagic transformation and mortality in our cohort of stroke patients. Establishing any causal link for these relationships needs further research. PMID- 19546542 TI - Normal magnetic resonance perfusion-weighted imaging in lacunar infarcts predicts a low risk of early deterioration. AB - BACKGROUND: Current clinical tools to identify lacunar infarct patients at risk of deterioration are inadequate, and imaging techniques to predict fluctuation and deterioration would be of value. We sought to determine the occurrence of MRI perfusion-weighted imaging (PWI) abnormalities in lacunes, and whether they help predict clinical and radiological outcome. METHODS: Patients with lacunar stroke or TIA were selected from a prospective MR imaging study. MRI was performed within 24 h of the event and follow-up imaging completed at 30 or 90 days. Baseline perfusion maps were qualitatively assessed and infarct volumes measured. Early clinical deterioration (NIHSS worsening of > or = 3 points within 72 h of event) and 90-day modified Rankin Scale score (mRS) were recorded. RESULTS: Twenty-two patients were included. Fifteen (68.2%) had abnormal PWI at the site of the diffusion-weighted imaging lesion. Patients with abnormal PWI were more likely to have stroke than TIA as their index event (RR 2.2, 95% CI 0.9-5.2, p = 0.02). Early clinical deterioration occurred in 4 patients (18.2%), all of whom had abnormal PWI. PWI lesions were not associated with a higher 90-day NIHSS or mRS score, nor did they predict infarct volume growth. CONCLUSIONS: MR-PWI abnormalities are seen in two thirds of lacunar infarcts, and are associated with stroke rather than TIA. Normal PWI identifies patients at low risk of early clinical deterioration. PMID- 19546543 TI - Surgical treatment of aortico-left ventricular tunnel: a 12-year experience. AB - OBJECTIVES: This study reviews 9 patients with aortico-left ventricular tunnel (ALVT) and aims to establish the optimal surgical strategies for this exceptional and rare pathology. METHODS: 54,882 patients underwent open-heart surgery at the Fuwai Hospital between July 1996 and May 2008. Nine of these patients were diagnosed with ALVT. One patient died of acute heart failure before operation. The remaining 8 patients underwent operation. The mean age of these 8 patients was 22.5 years. Clinical, echocardiographic and surgical details were reviewed. RESULTS: At operation the diagnosis was confirmed, and then repaired by direct suture closure (n = 1), single patch closure of the aortic end of the ALVT (n = 4), or aortic valve replacement (n = 3). In 5 patients, the aortic opening of the tunnel was above the right coronary sinus of Valsalva while in the other 3 it was above the left coronary sinus. All patients recovered safely. There was 1 late death (from perivalvular leak). The remaining patients remain asymptomatic at a mean follow up of 30.1 months. CONCLUSIONS: ALVT is a rare cardiac entity that should be treated soon after the diagnosis is made. The specific surgical operation should be individualized based on unique cardiac anomaly of each patient. PMID- 19546544 TI - Hepatocellular cancer (HCC) contributes in a significant way to the worldwide burden of neoplastic diseases.. Preface. PMID- 19546545 TI - Hepatocellular carcinoma--epidemiological trends and risk factors. AB - Hepatocellular carcinoma (HCC) is the third most common cause of cancer-related deaths worldwide with about 600,000 patients dying from the disease annually. In 70-90%, HCC develops on the background of chronic liver cirrhosis or inflammation. Risk factors and etiologies vary among geographical regions. In regions with a high incidence the majority of cases are related to HBV and HCV hepatitis. In developed countries, in addition to virus-related HCC, high consumption of alcohol as well as non-alcoholic fatty liver disease often in the context of metabolic syndromes are the prevalent causes. Improvement in clinical management of patients with liver cirrhosis and the control of related complications are the key for the rising incidence of HCC. This review gives an overview on epidemiological trends and risk factors and their mechanisms involved in the hepatocarcinogenesis. Knowledge of these factors will help to improve current concepts for prevention, screening and treatment of this disease. PMID- 19546546 TI - Hypotheses on the role of transforming growth factor-beta in the onset and progression of hepatocellular carcinoma. AB - Hepatocellular carcinomas (HCCs) constitute a therapeutic challenge with mostly unfavorable outcome. This may reflect our incomplete understanding of disease pathogenesis, e.g. the elucidation of tumorigenic signaling pathways. Knowledge gathered hitherto focuses on genetic alterations that result in the loss of tumor suppressor functions as well as amplification and mutation of cancer genes. Further evidence points to a decisive role of cytostatic and apoptotic functions mediated on hepatocytes by transforming growth factor (TGF)-beta. These effects are critical for the control of liver mass with loss of TGF-beta activities resulting in hyperproliferative disorders and cancer. This concept is based on studies that describe a bipartite role of TGF-beta with tumor suppressor functions at early stages of liver damage and regeneration, whereas during cancer progression TGF-beta may turn from a tumor suppressor to a tumor promoter that exacerbates invasive and metastatic behavior. Consequently and most importantly, the oncogenic potential of recent therapeutic approaches against profibrogenic TGF-beta effects needs to be carefully delineated and a cancer therapy with specific targets disrupting the TGF-beta signaling cascade may be envisioned. In line with this concept, we and others found overexpression of TGF-beta antagonist Smad7 in the majority of HCC samples, providing a mechanism for hepatocytes to escape TGF-beta-dependent growth control in the process of cancerogenesis. PMID- 19546547 TI - Hepatocellular carcinoma: pathology and liver biopsy. AB - Hepatocellular carcinoma (HCC) is the most frequent malignant tumor of the liver. The diagnosis of HCC depends on the measurement of serum alpha-fetoprotein levels and imaging techniques. Liver biopsy is recommended in uncertain cases. We review the morphological characteristics of HCC and their differential diagnoses including other liver tumors and metastases. We further specify problems and pitfalls in the differential diagnosis and discuss limitations of liver biopsies in the diagnosis of HCC. PMID- 19546548 TI - Transabdominal ultrasound with echoenhancement by contrast media in the diagnosis of hepatocellular carcinoma. AB - According to the European Association for the Study of the Liver (EASL) and the American Association for the Study of Liver Diseases (AASLD), ultrasound (US) is the recommended tool for surveillance of patients at risk of developing hepatocellular carcinoma (HCC). Larger HCCs can be diagnosed with a high accuracy by conventional US. However, the differentiation of smaller malignant lesions in cirrhotic livers can be improved by contrast-enhanced ultrasound (CEUS). Second generation contrast agents consisting of microbubbles enable us to visualize specific tumor vascularization patterns. With CEUS, it is not only possible to detect and characterize HCC nodules, but to control the effects of ablation techniques of HCC as well, evaluating the former lesion with respect to complete necrosis or residual viable tumor. Limitations of CEUS are its inability to characterize lesions distant to the applicator. Moreover, so far the use of contrast agents in US did not result in increased sensitivity in the detection of small HCCs (<1 cm). Thus, there is currently no indication to use contrast agents to increase the detection rate of HCC in patients undergoing US surveillance. PMID- 19546549 TI - Imaging of hepatocellular carcinoma by computed tomography and magnetic resonance imaging: state of the art. AB - Hepatocellular carcinoma (HCC) is a very frequent tumor worldwide. Its incidence is linked to the distribution of liver cirrhosis and viral hepatitis, which are the main risk factors for the development of HCC. For the evaluation of the cirrhotic liver and for the diagnosis of HCC, multidetector computed tomography (MDCT) proved to be a robust and reliable tool. In MDCT the diagnosis of HCC can be made based on neovascularization with increased arterial and decreased portal venous supply. With modern magnetic resonance imaging (MRI), spatial resolution and robustness increased dramatically. Beside the evaluation of neovascularization by means of gadolinium-enhanced early dynamic MRI, the main advantages of MRI are additional information on tissue composition and liver specific function. With diffusion-weighted imaging or plain T(1)- and T(2) weighted sequences, different tissue elements like fat, hemorrhage, glycogen, edema and cellular density can be evaluated. Liver-specific contrast agents give insight into the Kupffer cell density or the hepatocellular function. The integration of all these parts into the MR examination allows for a very high detection rate for overt HCC nowadays, although very small HCCs are still a challenge. Moreover, insight into the different stages of hepatocarcinogenesis can be possible with MRI. Despite its limited availability in some countries, it has to be rendered to be the modality of choice for the distinct evaluation of the cirrhotic liver. PMID- 19546550 TI - HCC diagnosis with liver-specific MRI--close to histopathology. AB - Thanks to the sensible and continuous improvements achieved, magnetic resonance imaging (MRI) can nowadays be considered the most accurate modality to image the liver. Moreover, the technique is the only one able to provide at the same time information about intracellular and vascular changes occurring in parenchymas. For these reasons, MRI plays a major role in the surveillance and follow-up of patients with cirrhosis. If a baseline MR study investigates the progressive alteration of lesion architecture, grading, stromal component, as well as intracellular content of fat, glycogen, or metal ions, thus leading to a frequent confident diagnosis of lesion nature, a dynamic study provides additional information about lesion vascular enhancement, which may represent the only clue for the differential diagnosis between premalignant and malignant lesions. In addition, the introduction of hepatobiliary contrast agents has further implemented the diagnostic confidence of the technique, permitting to explore the so-called grey area in which significant histological changes are already present without an evident arterial supply of the nodule. Although in the evaluation of liver pathologies MRI is mainly applied in the study of cirrhosis, the technique also plays a fundamental role in the assessment of other primitive liver malignancies, such as fibrolamellar carcinoma or cholangiocarcinoma. In these cases in particular, MRI is required to pose a differential diagnosis with other liver malignancies (such as metastases), and, once the nature of the neoplasm is assessed, to give an accurate locoregional staging. PMID- 19546551 TI - Clinical staging of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common cancers in the world. The resistance of HCC to existing treatments and the lack of biomarkers for early detection make it one of most hard-to-treat cancers. Surgical tumor resection, including liver transplantation, remains the only curative modality for HCC. Several clinical prognostic models have been developed for the staging of patients with HCC, but there is no general consensus which is the most reliable. There are restrictions in the use of these prognostic scores, because most scores were not validated in different countries and the HCCs differ in their biological behavior. A further problem is the heterogeneity of patients with HCC (with or without cirrhosis, compensated or decompensated cirrhosis and treated or nontreated patients); many studies combine patients with and without cirrhosis and patients with compensated and decompensated cirrhosis. The clinical efficiency of these scores is therefore limited to patients with HCC. Nowadays, DNA microarray technology has revolutionized the understanding of the molecular basis of HCC; therefore, many studies aimed to discover biomarkers for cancer staging, for prediction of recurrence, for prognosis, and for treatment selection. Most of the studies are too small for the development of predictors and at the moment microarray technology is too expensive for daily clinical use. Therefore, DNA microarray technology is at the moment not an established method in the prediction of the prognosis of patients with HCC. Future prognostic studies should include different predictors and should be performed in selected patient populations to determine whether specific prognostic indicators are more relevant at different stages of liver disease. PMID- 19546552 TI - Hepatocellular cancer: optimal strategies for screening and surveillance. AB - Hepatocellular carcinoma (HCC) is a challenging malignancy of global importance and is associated with a high rate of mortality. Individuals with chronic viral hepatitis and other forms of liver disease are at risk for developing HCC. The stage of cancer dictates the therapeutic choice, making early detection a primary objective. Many observational studies have reported that HCC is diagnosed at an earlier stage in patients who received surveillance. The guidelines of the American Association for the Study of Liver Diseases suggest that surveillance should be performed using ultrasonography at an interval of every 6-12 months. This interval is based upon mainly observational data and the expected growth rates of HCC. Patients with abnormal screening tests require additional investigation using computed tomography scanning, magnetic resonance imaging, or liver biopsy. Although the optimal methods of screening and the cost effectiveness of surveillance for HCC remain to be established, systematic screening still offers the best hope for early diagnosis, treatment eligibility, and improved survival. PMID- 19546553 TI - Local ablative therapies in HCC: percutaneous ethanol injection and radiofrequency ablation. AB - Hepatocellular carcinoma (HCC) is the most common primary hepatic malignancy. Surgery and transarterial chemoembolization have for many years dominated the local treatment of HCC. The introduction of image-guided percutaneous techniques for local tumor ablation changed the treatment of liver cancer. Percutaneous ethanol injection (PEI) and radiofrequency ablation (RFA) have both successfully been employed in the treatment of HCC. In current guidelines both techniques are recommended as standard therapy in limited liver cancer. From the current literature both techniques have to be considered safe and effective in the treatment of HCC. Several randomized trials showed RFA to be superior to PEI for treating small HCC. Moreover, several studies analyzed survival after PEI or RFA in comparison with surgery, proving interventional therapy to be a serious competitor of resection. With introduction of combined interventional therapies including transarterial chemoembolization, PEI and RFA survival was improved further. In this article, we provide an insight into the technical basics of PEI and RFA and review the clinical results and indications of these interventional techniques in the treatment of HCC. PMID- 19546554 TI - Transcatheter chemoembolization of unresectable hepatocellular carcinoma: current knowledge and future directions. AB - Transarterial chemoembolization (TACE) is considered the gold standard for the treatment of unresectable hepatocellular carcinoma (HCC) in patients with a preserved liver function and in the absence of clear extrahepatic tumor spread. This review article deals with the radiological and clinical workup, the interventional technique of chemoembolization for HCC and gives an overview of the common as well as some rare and dramatic complications of TACE. Finally, some new trends and advances in clinical research with regard to the interventional management of HCC are discussed. PMID- 19546555 TI - Treatment of hepatocellular carcinoma by radioembolization using 90Y microspheres. AB - The use of external beam radiation therapy for primary treatment of hepatocellular carcinoma (HCC) has been limited by the low radiation tolerance of the non-tumoral liver. However, technical advances allowing partial liver volume external irradiation have resulted in consistently high response rates. Internal radiation therapy, also called (90)Y radioembolization ((90)Y-RE), consists in delivering implantable microspheres labeled with (90)Y into the arteries that feed liver tumors in order to provide a high dose of radiation to tumor nodules irrespective of their number, size and location, while preserving the non-tumoral liver tissue from receiving a harmful level of radiation. Among patients with HCC, (90)Y-RE is used for those that have a preserved liver function and unresectable tumors that cannot be treated with percutaneous ablation. Although (90)Y-RE is by and large well tolerated, it may produce relevant toxic effects as a result of radiation of non-target organs including cholecystitis, gastrointestinal ulceration, pneumonitis, and most importantly, liver toxicity. A significant effect on tumor growth in the treated lesions is consistently observed with disease control rates in excess of 80%. Also, (90)Y-RE may allow downstaging large or multiple lesions to radical treatments with curative intent. When compared with the survival of HCC patients in advanced stage either not treated or treated with ineffective systemic agents, survival after (90)Y-RE is encouraging and warrants future clinical trials. Clinical research in combining the cytotoxic effect of (90)Y with the cytostatic mechanism of targeted therapies is currently in progress and will provide valuable safety and toxicity data that may translate into improved clinical outcome and overall survival. PMID- 19546556 TI - Image-guided interstitial high-dose-rate brachytherapy in hepatocellular carcinoma. AB - New interventional options especially for patients with HCC and BCLC scores B and C give rise to disputes about the optimal therapeutic management. CT-guided brachytherapy complements established interventional techniques like RFA and TACE since it may also be used successfully in tumors much greater than 5 cm in diameter. In addition, unlike thermal ablation, the brachytherapy technique may be applied in tumors located nearby risk structures such as liver hilum or gallbladder and it is independent of cooling effects such as through large blood vessels or strong tumor perfusion. Depending on tumor size, geometry and visibility, MRI or CT guidance may be used. 15 Gy minimal target dose can be applied safely in a single or--in case of very large tumors--a sequential approach targeting different tumor portions. Local recurrence rates will be very low, and the rate of complications is moderate despite the fact that most patients present with underlying cirrhosis and related comorbidities. Preliminary data suggest a positive impact on overall survival. Randomized controlled trials are on their way to assess combination schemes with systemic treatments such as sorafenib. PMID- 19546557 TI - Systemic therapies in hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common and lethal malignant tumors worldwide in the human population. Due to late diagnosis and/or advanced underlying liver cirrhosis, only limited treatment options with marginal clinical benefit are available in up to 70% of patients. During the last decades, no effective conventional cytotoxic systemic therapy was available contributing to the dismal prognosis in patients with advanced disease. However, a better knowledge of molecular hepatocarcinogenesis provides today the opportunity for targeted therapy. Positive data from the pivotal phase III SHARP trial assessing the efficacy and safety of the multikinase inhibitor sorafenib broadened the horizon for patients with advanced disease. After years of therapeutic nihilism, sorafenib was the first agent to demonstrate a statistically significant improvement in overall survival for patients with advanced HCC. This article reviews the historical perspective of systemic therapy in HCC and provides a brief overview of molecular hepatocarcinogenesis and potential targets in HCC. Most promising molecular targeted agents tested within clinical trials in advanced HCC are summarized, with a special attention to sorafenib, sunitinib, bevacizumab, and erlotinib. PMID- 19546558 TI - Hepatocellular carcinoma and liver cirrhosis: assessment of the liver function after Yttrium-90 radioembolization with resin microspheres or after CT-guided high-dose-rate brachytherapy. AB - PURPOSE: To identify changes of liver function after single-fraction irradiation or yttrium-90 radioembolization ((90)Y-RE) of hepatocellular carcinoma associated with liver cirrhosis on the basis of laboratory data. METHODS AND MATERIALS: 24 patients with primary liver carcinoma and liver cirrhosis classified Child-Pugh A or B were treated either by image-guided high-dose-rate brachytherapy (HDR-BT) (12 patients) or by (90)Y-RE (12 patients). The following laboratory parameters were assessed 1 day before and 3 days, 6 weeks and 3 months after the intervention: total bilirubin and gamma-glutamyl transpeptidase (GGTP) as parameters of detoxification function, albumin and cholinesterase (ChE) as direct synthesis parameters, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (AP) as indicators of liver tissue damage. Preinterventional values were taken as baseline, following values were calculated as percentage changes from the baseline value. Statistical analysis was performed using the Wilcoxon-matched pairs test, comparing postinterventional with preinterventional values. Differences were considered statistically significant with a p value <0.05. RESULTS: In all patients the median bilirubin, ALT, AP and albumin values remained within normal limits at any time of follow-up. AST levels in the RE group and GGTP in both groups have been already elevated over a normal range before the intervention, and in both groups both parameters showed a slight increase after interventions. ChE activity was lowered already in the baseline values and showed a further decrease 3 days after BT as well as 3 days and 6 weeks after RE, with final reconstitution to baseline values. All liver function test parameters showed mild changes shortly after radiation therapy but floating laboratory values recovering within 12 weeks to baseline values. Radiation or RE induced liver disease was recorded in no patient. CONCLUSIONS: Liver function parameters show only mild changes shortly after intervention with recovery within 6-12 weeks to baseline values. PMID- 19546559 TI - Pro-inflammatory cytokine genes influence the clinical features of frontotemporal lobar degeneration. AB - BACKGROUND/AIMS: Recent studies suggested a role for pro-inflammatory mediators in frontotemporal lobar degeneration (FTLD). The objective of this study was to evaluate the association of functionally active polymorphisms in pro-inflammatory cytokine genes with the occurrence and the clinical features of the disease. METHODS: Using a case-control study, we compared allelic and genotypic frequencies of several polymorphisms in the interleukin (IL)-1alpha, interleukin (IL)-1beta, interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha genes between 110 FTLD patients and 119 healthy controls. RESULTS: No significant association between the examined polymorphisms and the disease was found. However, in comparison with remaining genotypes, patients carrying the T/T genotype of the IL-1beta gene showed a significantly lower age at onset of the disease. In addition, scores of the Frontal Assessment Battery were significantly modified by the IL-6 -174G>C polymorphism. CONCLUSION: Our findings support a role for pro-inflammatory cytokine genes in the pathogenesis of frontotemporal lobar degeneration. PMID- 19546561 TI - Development of mechanisms that regulate global CNS states: an introduction. PMID- 19546562 TI - Arousal-related reticular neurons during reduced oxygen tension: resilience and recovery of electrical activity. AB - Whole-cell patch clamp recordings of the electrical activity of large medullary reticular formation neurons, in nucleus gigantocellularis, were performed under control conditions and under conditions of hypoxia or anoxia. Neurons were discovered whose activity was remarkably resilient during and after the reduction or loss of oxygen. Such cells may relate to the ability of the newborn brain to survive hypoxia/anoxia, and also may demonstrate the preservation of neurons involved in generalized CNS arousal, as would be appropriate for activating behavioral responses to the reduction or loss of oxygen. PMID- 19546563 TI - Neonatal EEG/sleep state analyses: a complex phenotype of developmental neural plasticity. AB - Computer analyses of EEG/sleep states can be used as physiologic biomarkers of developmental neural plasticity. Frequency- and time-dependent signal processing strategies of cerebral and noncerebral measures can help test current theories of neuronal network maturation in terms of segregation and integration of short distance versus long-distance neuronal connections throughout the neuroaxis. Specific phenotypic expressions of adaptive or maladaptive neuronal connectivity are proposed based on comparisons of whole-brain EEG/sleep resting states between preterm and full-term cohorts when developmental outcome measures are applied. Combined use of neurophysiological datasets with neuroimaging and genetic methodologies define endophenotypes that will more accurately diagnose children at risk for developmental disorders, as well as design appropriate neuroprotective interventions for the individual's age and disease progress. PMID- 19546560 TI - Diagnosis and management of non-erosive reflux disease--the Vevey NERD Consensus Group. AB - BACKGROUND/AIMS: Although considerable information exists regarding gastroesophageal reflux disease with erosions, much less is known of non-erosive reflux disease (NERD), the dominant form of reflux disease in the developed world. METHODS: An expert international group using the modified Delphi technique examined the quality of evidence and established levels of agreement relating to different aspects of NERD. Discussion focused on clinical presentation, assessment of clinical outcome, pathobiological mechanisms, and clinical strategies for diagnosis and management. RESULTS: Consensus was reached on 85 specific statements. NERD was defined as a condition with reflux symptoms in the absence of mucosal lesions or breaks detected by conventional endoscopy, and without prior effective acid-suppressive therapy. Evidence supporting this diagnosis included: responsiveness to acid suppression therapy, abnormal reflux monitoring or the identification of specific novel endoscopic and histological findings. Functional heartburn was considered a separate entity not related to acid reflux. Proton pump inhibitors are the definitive therapy for NERD, with efficacy best evaluated by validated quality-of-life instruments. Adjunctive antacids or H(2) receptor antagonists are ineffective, surgery seldom indicated. CONCLUSIONS: Little is known of the pathobiology of NERD. Further elucidation of the mechanisms of mucosal and visceral hypersensitivity is required to improve NERD management. PMID- 19546565 TI - Prenatal stress and neurodevelopment of the child: focus on the HPA axis and role of the placenta. AB - Recent human studies have shown that a wide variety of prenatal stressors, from anxiety and partner relationship problems, to natural disasters, increase the risk for a diverse range of adverse neurodevelopmental outcomes in the child. These include impaired cognitive development and behavioral problems, autism and schizophrenia. However, many questions remain about the underlying processes. Much of the research, based on animal studies, has focussed on the maternal HPA axis, with mixed results. Maternal stress or anxiety during pregnancy has been found to be weakly associated with raised maternal cortisol, if at all. The placenta may be a more promising programming vector, because it controls fetal exposure to the maternal environment. Animal studies indicate that prenatal stress can affect the activity of the placental barrier enzyme 11-betaHSD2, which metabolises cortisol. We review the evidence for a similar mechanism in humans and how maternal stress may cause other changes in the placenta which affect fetal neurodevelopment. PMID- 19546564 TI - Adolescent changes in the homeostatic and circadian regulation of sleep. AB - Sleep deprivation among adolescents is epidemic. We argue that this sleep deprivation is due in part to pubertal changes in the homeostatic and circadian regulation of sleep. These changes promote a delayed sleep phase that is exacerbated by evening light exposure and incompatible with aspects of modern society, notably early school start times. In this review of human and animal literature, we demonstrate that delayed sleep phase during puberty is likely a common phenomenon in mammals, not specific to human adolescents, and we provide insight into the mechanisms underlying this phenomenon. PMID- 19546567 TI - Individual differences in behavioral, physiological, and genetic sensitivities to contexts: implications for development and adaptation. AB - Although exposure to adversity places children at high risk for developmental problems, there is considerable variation in the adaptation of children exposed to both low and high levels of adversity. In recent years, researchers have made significant progress in understanding how social environments shape children's development. Studies indicate that not all children are equally susceptible to environmental effects. In this article, we review the studies that examine individuals' sensitivity to both positive and negative contextual influences at three levels of analysis: behavioral, physiological, and genetic. Furthermore, we examine how these different types of sensitivities may be related and whether early exposure to adversity plays an important role in shaping development of individual reactivity to contexts. We also suggest important directions for future studies of individual differences in susceptibility to environmental effects. PMID- 19546566 TI - Prefrontal plasticity and stress inoculation-induced resilience. AB - Coping with mild early life stress tends to make subsequent coping efforts more effective and therefore more likely to be used as a means of arousal regulation and resilience. Here we show that this developmental learning-like process of stress inoculation increases ventromedial prefrontal cortical volumes in peripubertal monkeys. Larger volumes do not reflect increased cortical thickness but instead represent surface area expansion of ventromedial prefrontal cortex. Expansion of ventromedial prefrontal cortex coincides with increased white matter myelination inferred from diffusion tensor magnetic resonance imaging. These findings suggest that the process of coping with early life stress increases prefrontal myelination and expands a region of cortex that broadly controls arousal regulation and resilience. PMID- 19546568 TI - Role of attention in the regulation of fear and anxiety. AB - Emotion regulation makes use of specific aspects of attention and executive functions that are critical for the development of adaptive social functioning, and perturbations in these processes can result in maladaptive behavior and psychopathology. Both involuntary and voluntary attention processes have been examined at both the behavioral and the neural levels and are implicated in the maintenance of fearful or anxious behaviors. However, relatively little is known about how these attention processes come to influence emotional behavior across development. The current review summarizes the extant literature on the links between voluntary and involuntary attention processes and the role that these attention processes have in the etiology, maintenance, and regulation of anxious behavior. PMID- 19546571 TI - Hormones and sexual differentiation of avian social behavior. AB - Research on the hormonal basis of sexual differentiation of mammalian behavior and its neural substrates constitutes an important chapter in the study of neural development that has led to important insights into how to conceptualize sexuality. Here research on sexual differentiation of avian social behavior is discussed with an eye toward potential insights into avian sexuality. Hormone manipulation experiments with Japanese quail (Coturnix japonica) and zebra finches (Taeniopygia guttata) reveal dissociations between different components of sexual and social behavior. None of the components of the male behavioral phenotype appear to be organized by androgens. Avian sexuality arises from multiple processes and pathways that appear to differ for mating behavior, sexual partner preference, and pairing. This provides the mechanistic bases for changes in social behavior in the course of avian evolution. PMID- 19546569 TI - The meaning of weaning: influence of the weaning period on behavioral development in mice. AB - Maternal care during the first week postpartum has long-term consequences for offspring development in rodents. However, mother-infant interactions continue well beyond this period, with several physiological and behavioral changes occurring between days 18 and 28 PN. In the present study, we investigate the long-term effects on offspring behavior of being weaned at day 21 PN versus day 28 PN. We found that male and female offspring engage in higher initial levels of social interaction if weaned at day 28 PN, as well as sexually dimorphic changes in exploratory behavior. Females who were themselves weaned earlier also appeared to wean their own pups earlier. Sex-specific effects of weaning age were found on levels of oxytocin and vasopressin V1a receptor density in the hypothalamus, central nucleus of the amygdala and nucleus accumbens. These results indicate that altering weaning age in mice may be a useful model for investigating the development of sexual dimorphism in neurobiology and behavior. PMID- 19546570 TI - Consequences of early experiences and exposure to oxytocin and vasopressin are sexually dimorphic. AB - In the socially monogamous prairie vole, we have observed that small changes in early handling, as well as early hormonal manipulations can have long-lasting and sexually dimorphic effects on behavior. These changes may be mediated in part by changes in parental interactions with their young, acting on systems that rely on oxytocin (OT) and arginine vasopressin (AVP). Knowledge of both endogenous and exogenous influences on systems that rely on OT and AVP may be helpful in understanding sexually dimorphic developmental disorders, such as autism, that are characterized by increased anxiety and deficits in social behavior. PMID- 19546572 TI - Is there a genomic tone? Implications for understanding development, adaptation and treatment. AB - This review examines the general hypothesis that cellular plasticity may be related to genomic plasticity. Using a novel approach to microarray analysis, recent studies from our laboratory showed that, in genetically identical mice, variability in the large-scale organization of gene expression was associated with individual differences in behavior. An association of the large-scale organization of gene expression in the hippocampus with behavioral variability suggests that an important aspect of gene expression regulation had gone unrecognized with traditional genomic approaches designed to look for specific differences in gene expression levels. Here we introduce the concept of a 'genomic tone' as a global genomic property related to the levels and balance between factors that generally promote and factors that generally inhibit gene expression. We suggest that genomic tone may be related to the plasticity of the gene expression system and may be an important target for developmental and therapeutic interventions. PMID- 19546573 TI - Antimesenteric tapering enteroplasty in an adult patient with clinical evidence of obstruction: a bowel-preserving alternative to resection. AB - BACKGROUND/AIMS: The approach to small bowel obstruction, when unrelieved with medical management, is open or laparoscopic exploration, often necessitating lysis of adhesions and the resection of any compromised bowel. In patients with prior bowel resections, the surgeon must tread a fine line, so as not to precipitate the clinical derangement known as short bowel syndrome (SBS). Techniques have been described that curtail the extent of intestinal resection, but these are primarily limited to the pediatric literature and are not commonly practiced in the general surgical population. METHODS: We report a case of a complicated small bowel obstruction in a patient with pre-existing short bowel length, in which a tapering enteroplasty was performed. RESULTS: Antimesenteric tapering successfully achieved a return of bowel function, avoiding the morbidity of an extended small bowel resection and the possibility of developing SBS. CONCLUSION: In such patients who are at high risk of developing SBS, in whom a segment of dilated small bowel has become defunctionalized, leading to significant and life-threatening symptoms, this procedure has the potential to help prevent SBS and its lifelong complications and associated mortality. PMID- 19546574 TI - Tree hole utilisation by the hairy-eared dwarf lemur (Allocebus trichotis) in Analamazaotra Special Reserve. AB - In this study we describe tree hole characteristics and use by the hairy-eared dwarf lemur (Allocebus trichotis) to determine habitat needs, potential functions of tree holes and sleeping group composition. We radio-tracked 6 adult individuals between April and November 2007 in the Analamazaotra Special Reserve. Tree holes were 1-9 m high (median: 7 m), in living trees measuring 26-54 cm in diameter at breast height (median: 32 cm), and could be a limiting resource. Each individual used 4 or 5 tree holes and had high nest fidelity. Animals most often slept socially in mixed-sex groups of 2-6 individuals and occasionally shared a tree hole with white-tailed tree rats (Brachytarsomys albicauda). We identified two sleeping groups: one composed of 2 adult males, 2 adult females and 2 juveniles; one composed of at least 2 adult females and 2 juveniles. Although tree holes were generally group exclusive, some intergroup sleeping was observed. Tree holes could have antipredator and thermoregulatory functions. Further research into sleeping hole availability, nest use and the degree of niche separation or competition between sympatric Cheirogaleidae and other tree hole users (e.g. endemic rodents) is needed to assess better the conservation needs of these species. PMID- 19546575 TI - Virological and biochemical features in elderly HCV patients with hepatocellular carcinoma: amino acid substitutions in HCV core region as predictor of mortality after first treatment. AB - AIMS: We showed previously that amino acid (aa) substitutions in HCV genotype 1b (HCV-1b) core region are negative predictors of virological response to peginterferon + ribavirin therapy, and also risk factors of hepatocarcinogenesis. The aim of this study was to evaluate the impact of core aa substitutions on mortality in elderly patients. METHODS: We compared the characteristics and survival of 92 elderly (> or =75 years) patients with HCV-related hepatocellular carcinoma (HCC) (including 62 patients with HCV-1b) with those of 44 younger patients (<50 years, 34 patients with HCV-1b). RESULTS: For all patients, univariate analysis identified female sex, history of blood transfusion, preserved liver function and glucose metabolism as significant variables in the elderly patients. In patients with HCV-1b-related HCC, univariate analysis identified preserved lipid metabolism as significant variable in addition to significant variables in overall patients. In elderly patients with HCV-1b related HCC, multivariate analysis identified male sex, methionine of core aa91, and non-radical therapy as factors that influenced mortality after first treatment for HCC. CONCLUSIONS: Our results characterized elderly patients who develop HCC after HCV-1b infection, and suggested that aa substitutions of HCV-1b core region correlate with mortality of patients after first treatment for HCC. PMID- 19546576 TI - FK778 attenuates cytomegalovirus-enhanced vein graft intimal hyperplasia in a rat model. AB - BACKGROUND: Venous grafts are commonly used to treat drug-resistant coronary artery disease, although long-term functionality is limited because of proliferation and migration of smooth muscle cells (SMC). As proliferating SMC are particularly susceptible for the stimulating effects of cytomegalovirus (CMV), we hypothesized that CMV infection may enhance cell proliferation and graft failure. Furthermore, we evaluated the potential of FK778 to prevent intimal hyperplasia. Apart from its antiviral properties, FK778 is a new immunosuppressive agent which may also affect SMC proliferation, making it an interesting drug to prevent (CMV-enhanced) venous graft intimal hyperplasia. METHODS: Epigastric vein-to-common femoral artery interposition grafts were placed in four groups of 10 rats each. Rats received either FK778 (oral treatment, 15 mg/kg), were infected with CMV (1.25 x 10(6) plaque-forming units) or were both treated and infected. RESULTS: CMV infection resulted in a significant increase in intimal and medial cross-sectional area and medial wall thickness of the vein grafts. This effect was diminished by administration of FK778. Moreover, FK778 treatment alone resulted in a significant decrease in neointimal area and percentage of stenosis versus the control group. CONCLUSIONS: These data suggest a role of CMV in venous graft failure. Also, our results suggest a prospective role for the new immunosuppressive drug FK778 in the prevention of (CMV-mediated) vein graft intimal hyperplasia. PMID- 19546577 TI - Sequence identity between the genomes of humans and viruses. AB - OBJECTIVE: People exposed to viruses usually do not develop diseases. Viruses cause diseases; however, most of the pathologic effects of these diseases are not caused directly by the viruses but by the side effects of the immune response. It is possible that sequence identity may exist between viruses and humans and may be biologically relevant to the infection of virus. The aim of this study was to find out whether sequence identity exists between viruses and humans. METHODS: BLAST searches of virus genome sequences against human genome were performed using the resources of the National Center for Biotechnology Information (NCBI). RESULTS: All the viruses that were compared to humans contain different numbers of short identical regions to the human genome. Each identical region was 20-30 nt and some were found at multiple sites within the human genome. CONCLUSION: A new role of the virus in the relationship between viruses and humans was taken into account. We assume that viruses have brought their sequence into the human genome in the long past history of human-virus interaction. In the process, viruses play roles as carriers to transfer external sequence information into the human genome. PMID- 19546578 TI - Is it important to model the impact of blood flow on the dose of drugs delivered transcutaneously? PMID- 19546580 TI - Genetics of diabetic nephropathy: are there clues to the understanding of common kidney diseases? AB - Diabetic nephropathy is the most common cause of end-stage renal disease in the Western world. There is evidence for a genetic susceptibility to diabetic kidney disease, but despite intensive research efforts it has proved difficult to identify the causative genes. Improvements in genotyping technologies have made genome-wide association studies (GWAS), employing hundreds of thousands of single nucleotide polymorphisms, affordable. Recently, such scans have advanced understanding of the genetics of common complex diseases, finding more than 100 novel susceptibility variants for diverse disorders including type 1 and 2 diabetes, coronary heart disease, Crohn's disease and rheumatoid arthritis. In this review, type 2 diabetes is highlighted to illustrate how genome-wide association studies have been used to study the genetics of complex multifactorial conditions; in addition, diabetic nephropathy will be used to demonstrate how similar scans could be employed to detect genetic factors predisposing to kidney disease. The identification of such variants would permit early identification of atrisk patients, enabling targeting of therapy and a move towards primary prevention. In addition, these powerful research methodologies may identify genes that were not previously known to predispose to nephropathy, thereby enhancing our understanding of the pathophysiology of renal disorders and potentially leading to novel therapeutic approaches. PMID- 19546581 TI - Renal replacement therapy for acute kidney injury. AB - The treatment of established acute kidney injury (AKI) is largely supportive in nature. Renal replacement therapy remains the cornerstone of management for the minority of patients who have severe AKI. Optimization of renal replacement therapy may modulate the high mortality associated with AKI. Recent trials indicated that continuous renal replacement therapy does not confer a survival advantage as compared to intermittent hemodialysis. Furthermore, there is no evidence to support a more intensive strategy of renal replacement therapy in the setting of AKI. There is comparatively limited data regarding the ideal timing of renal replacement therapy initiation and the preferred mode of solute clearance. PMID- 19546582 TI - Peroxisome proliferator-activated receptor gamma agonists in kidney disease- future promise, present fears. AB - The peroxisome proliferator-activated receptor superfamily (PPARs) comprises a class of nuclear receptors with significant effects in regulating multiple cellular pathways. Much research and clinical interest has surrounded the PPAR gamma isoform because of its key role in the transcriptional regulation of metabolic pathways and the efficacy of thiazolidinediones, the most clinically used PPAR-gamma agonist, in the management of type 2 diabetes mellitus. In this review, we discuss the pathogenic role of PPAR-gamma in experimental models of kidney disease, clinical trials of thiazolidinediones in diabetic and non diabetic kidney disease, recent safety concerns surrounding PPAR-gamma agonists and reflect on their potential use in 'orphan' kidney diseases. PMID- 19546579 TI - Evaluation of polymorphisms in paraoxonase 2 (PON2) gene and their association with cardiovascular-renal disease risk in Mexican Americans. AB - BACKGROUND/AIMS: Genetic polymorphisms in the paraoxonase 2 (PON2) gene are thought to alter its activity and contribute to the development of cardiovascular and renal disease risk. The purpose of this study is to determine whether the Arg148Gly, Cys311Ser and rs12794795 polymorphisms of PON2 examined previously by others, are associated with type 2 diabetes (T2DM), and subclinical measures of cardiovascular and renal disease risk in Mexican Americans. METHODS: Study participants (n = 848; 21 families) were genotyped for the three polymorphisms by TaqMan assay. Association between the genotypic and phenotypic data was performed by measured genotype approach as implemented in the variance component analytical tools. RESULTS: The Arg148Gly variant was found to be monomorphic in our dataset. Of the phenotypes examined for association, the A/C variant located in intron-1 (rs12794795) exhibited statistically significant association only with diastolic blood pressure (p = 0.018) after accounting for the trait-specific covariate effects. The Cys311Ser variant failed to show statistically significant association with any of the phenotypes examined. CONCLUSION: In conclusion, the variants examined at the PON2 locus in Mexican Americans do not appear to be a major contributor to T2DM, cardiovascular or renal disease risk, although they exhibited a small effect on the blood pressure values. PMID- 19546583 TI - Influence of dual blockade of the renin-angiotensin system on thirst in hemodialysis patients. AB - BACKGROUND/AIMS: Angiotensin II promotes sodium retention and influences the central regulation of fluid intake. Clinical studies on the effect of an angiotensin-converting enzyme inhibitor (ACEI) on xerostomia and thirst in chronic hemodialysis (HD) patients were scarce and gave contradictory results. We hypothesized that a more effective inhibition of the renin-angiotensin aldosterone axis with the combined ACEI and angiotensin receptor antagonist administration may reduce thirst and xerostomia,thereby decreasing interdialytic weight gain (IWG) in HD patients. METHODS: Twenty-one chronic HD patients (16 men, 5 women, mean age 54 +/- 13 years, time on dialysis 50 +/- 58 months) who had been on chronic ACEI therapy were studied. In a double-blind, crossover study, all subjects received in a random order either losartan (50 mg/day) or placebo for two 4-week periods with a 7-day wash-out. Basic biochemistry, serum electrolytes, plasma aldosterone, measurements of salivary flow rate after stimulation with paraffin chewing, and subjective xerostomia and thirst questionnaires were collected before dialysis sessions both before and after each treatment period. IWG and blood pressure were assessed at each dialysis. RESULTS: The volume of saliva increased after losartan (from 1.2 +/- 0.7 to 1.5 +/- 1.0 ml/min, respectively; p = 0.03), but this was reflected neither by the severity of the symptoms of xerostomia assessed by the patients on a visual analog scale (31 +/- 9 vs. 31 +/- 8 mm, respectively) nor by the intensity of thirst (final score 22 +/- 5 vs. 21 +/- 5 at baseline). No changes of the mean IWG were observed during the treatment with losartan (2.5 +/- 0.6 kg before and 2.4 +/- 0.8 kg at the end of the treatment). Plasma aldosterone decreased at the end of the losartan therapy (from 151 +/- 86 to 111 +/- 51 pg/ml; p = 0.02). Predialysis serum potassium did not change during the study. CONCLUSIONS: The addition of an angiotensin receptor blocker to chronic ACEI therapy is not effective in reducing thirst and thereby IWG in chronic HD patients. PMID- 19546584 TI - Effect of hemodialysis on acid leukocyte-type ribonuclease, alkaline ribonuclease and polymorphonuclear elastase serum levels in patients with end-stage renal disease. AB - BACKGROUND: Low-molecular-weight proteins (LMWPs) are substances of molecular weights 10-35 kDa, which accumulate in plasma of patients with end-stage renal disease (ESRD) due to the abolishment of plasma renal filtration. LMWPs are considered as a separate group of uremic toxins. AIM: The influence of hemodialysis (HD) on the release of some LMWPs from leukocytes was assessed by comparing levels of serum pancreatic-type alkaline RNase and leukocyte-type acid RNase as well as polymorphonuclear (PMN) elastase. METHODS: The mentioned proteins were assayed in 58 ESRD patients on HD prior and after the dialysis session and compared with the results obtained from 36 healthy subjects. The levels of elastase and acid and alkaline RNase were correlated with HD parameters, residual diuresis, predialysis concentrations of serum creatinine, urea and albumin as well as pre- and postdialysis granulocyte count. RESULTS: Changes in PMN elastase produced by the dialysis session positively correlate with changes in acid RNase levels (r = 0.3650; p = 0.0061), while there is no such correlation for alkaline RNase. There is a negative correlation between pre- and postdialysis differences in levels of acid and alkaline RNases (r = -0.3542; p = 0.008), indicating that HD induces liberation of a factor suppressing alkaline RNase. Levels of acid and alkaline RNase negatively correlate with residual diuresis, indicating its significance in control of LMWP accumulation (r = -0.3970; p = 0.0025; r = -0.2596; p = 0.0533, respectively). CONCLUSIONS: Dialysis treatment causes an increase in both acid leukocyte-type and alkaline pancreatic-type RNase activity in plasma. Dialysis-related increases in acid RNase activity correlate with the respective changes in PMN elastase, which suggests that leukocyte activation during dialysis contributes to an increase in plasma LMWPs. PMID- 19546585 TI - Impact of residual renal function on plasma levels of advanced oxidation protein products and pentosidine in peritoneal dialysis patients. AB - BACKGROUND: Residual renal function (RRF) affects the survival rate and the development of cardiovascular disease in peritoneal dialysis (PD) patients. We evaluated the impact of RRF on oxidative and carbonyl stress in PD patients. METHODS: Plasma advanced oxidation protein products (AOPP) and pentosidine were measured in PD patients with a urine volume of > or =300 ml/day (group A, n = 17) and <300 ml/day (group B, n = 14). AOPP and pentosidine were reevaluated after 12 months of follow-up in group A. RESULTS: Plasma levels of AOPP and pentosidine in group A were significantly lower than those in group B. Renal creatinine clearance was inversely correlated with AOPP (p < 0.05) and pentosidine (p < 0.01). After 12 months of follow-up, no significant change was observed in AOPP and pentosidine in groups who maintained a urine volume of > or =300 ml/day, but significantly increased in groups whose urine volume decreased to less than 300 ml/day. There were significant inverse relationships between the changes in renal creatinine clearance and AOPP (p < 0.01) and pentosidine (p < 0.05). CONCLUSION: Loss of RRF is associated with increased plasma AOPP and pentosidine, indicating that preservation of RRF has a beneficial effect in reducing the oxidative and carbonyl stress in PD patients. PMID- 19546586 TI - Truncating mutations in the chloride/proton ClC-5 antiporter gene in Seven Jewish Israeli families with Dent's 1 disease. AB - Dent's disease is an X-linked hereditary renal tubular disorder characterized by low-molecular-weight proteinuria (LMWP), hypercalciuria, nephrocalcinosis, nephrolithiasis, rickets and progressive renal failure. About 60% of patients have mutations in the CLCN5 gene (Dent 1), which encodes a kidney-specific chloride/proton antiporter, and 15% of patients have mutations in the OCRL1 gene (Dent 2). The aim of the study was to identify CLCN5 mutations in Jewish Israeli families with Dent's disease and to characterize the associated clinical syndromes. We studied 17 patients from 14 unrelated Israeli families with a clinical diagnosis of Dent's disease. LMWP was detected in all patients. Most of the affected individuals had hypercalciuria and nephrocalcinosis. Renal stones were found in 1 patient, and renal insufficiency developed in 2 patients. We identified six different truncating CLCN5 mutations that were segregated with the disease in 7 families: three nonsense mutations (Arg28stop, Arg467stop and Arg637stop), one deletion mutation (505delA) and two novel mutations, consisted of one deletion mutation (1493delG) and one insertion mutation (409insC). All the mutations cause premature termination of protein translation and result in a non functional truncated protein. The clinical characteristics of patients with different mutations were, in general, similar. PMID- 19546587 TI - The Impact of type 2 diabetes on mortality in end-stage renal disease patients differs between genders. AB - BACKGROUND/AIMS: In diabetics with end-stage renal disease (ESRD), risk of death has been reported to be non-constant after the first dialysis, and different outcomes have been observed between genders. We assessed the impact of type 2 diabetes (T2DM) on mortality in dialysis regarding its differential effect by gender using time-dependent analyses. METHODS: All T2DM and non-diabetic (no-DM) patients who started dialysis in two renal units in Lyon, France, between January 1, 1995, and December 31, 2007, were included. In multivariate analyses, the Cox model and Shoenfeld residual approach were used to assess the effect of T2DM on dialysis mortality by gender. RESULTS: We included 235 T2DM (males: 57.9%) and 480 no-DM (males: 65.6%) patients. In males, the adjusted hazard ratio (aHR) for death in T2DM versus no-DM was 0.83 (p = 0.20) and was constant over time after the first renal replacement therapy (RRT) (p = 0.88). In females, aHR for death in T2DM versus no-DM patients was not constant over time (p = 0.002). It was 0.64 (p = 0.13) within the first year after the first RRT and 2.10 (p = 0.002) after the first year. Evolutions with time of these aHR by gender were significantly different (p = 0.009). CONCLUSIONS: T2DM was associated with death only in females. This association was not constant over time after the first dialysis. PMID- 19546588 TI - Does increasing blood pH stimulate protein synthesis in dialysis patients? AB - BACKGROUND: Although the mechanism of muscle wasting in end-stage renal disease is not fully understood, there is increasing evidence that acidosis induces muscle protein degradation and could therefore contribute to the loss of muscle protein stores of patients on hemodialysis, a prototypical state of chronic metabolic acidosis (CMA). Because body protein mass is controlled by the balance between synthesis and degradation, protein loss can occur as result of either increased breakdown, impaired synthesis, or both. Correction of acidosis may therefore help to maintain muscle mass and improve the health of patients with CMA. We evaluated whether alkalizing patients on hemodialysis might have a positive effect on protein synthesis and on nutritional parameters. METHODS: Eight chronic hemodialysis patients were treated daily with oral sodium bicarbonate (NaHCO(3)) supplementation for 10-14 days, yielding a pre-dialytic plasma bicarbonate concentration of 28.6 +/-1.6 mmol/l. The fractional synthesis rates (FSR) of muscle protein and albumin were obtained by the L [(2)H(5)ring]phenylalanine flooding technique. RESULTS: Oral NaHCO(3 )supplementation induced a significant increase in serum bicarbonate (21.5 +/- 3.4 vs. 28.6 +/- 1.6 mmol/l; p = 0.018) and blood pH (7.41 vs. 7.46; p = 0.041). The FSR of muscle protein and the FSR of albumin did not change significantly (muscle protein: 2.1 +/- 0.2 vs. 2.0 +/- 0.5% per day, p = 0.39; albumin: 8.3 +/- 2.2 vs. 8.6 +/- 2.5% per day, p = 0.31). Plasma concentrations of insulin-like growth factor 1 decreased significantly (33.4 +/- 21.3 vs. 25.4 +/- 12.3 nmol/l; p = 0.028), whereas thyroid-stimulating hormone, free thyroxin and free triiodothyronine did not change significantly and nutritional parameters showed no improvement. CONCLUSION: In contrast to other findings, raising the blood pH of dialysis patients was not associated with a positive effect on albumin and muscle protein synthesis, or nutritional and endocrinal parameters. PMID- 19546589 TI - Lipopolysaccharide-triggered acute aggravation of mesangioproliferative glomerulonephritis through activation of coagulation in a high IgA strain of ddY mice. AB - BACKGROUND: The high IgA (HIGA) strain of ddY mice represents an inbred model of IgA nephropathy that shows mesangioproliferative glomerulonephritis with mesangial IgA deposition. In this study, aggravation of glomerulonephritis in HIGA mice through lipopolysaccharide (LPS)-triggered activation of coagulation was investigated. METHODS: Twelve-week-old HIGA and BALB/c mice were intraperitoneally injected with LPS twice at an interval of 3 days, and kidney specimens were collected 7 days after the second LPS injection. In an intervention experiment, the factor Xa inhibitor danaparoid was injected intraperitoneally every day for 7 days after the first LPS injection. RESULTS: LPS injection induced macrophage infiltration and cellular proliferation in the mesangium together with fibrin deposition and monocyte chemoattractant protein 1 mRNA expression, as well as antigen deposition of tissue factor, factor V, factor X, and protease-activated receptor 2. These phenomena were obvious in HIGA mice when compared to BALB/c mice. Interestingly, toll-like receptor 4 was intensely expressed in HIGA mice before LPS injection and subsequently decreased. Danaparoid treatment significantly ameliorated proteinuria, cellular proliferation, and fibrin deposition. CONCLUSIONS: The present data suggest that tissue factor and factor V induction by LPS may in part accelerate mesangioproliferative glomerulonephritis through activation of factor X and downstream proinflammatory and procoagulant mechanisms. PMID- 19546590 TI - Mesangial cells connected by the N-cadherin-catenin system in the rat kidney. AB - BACKGROUND/AIMS: Mesangial cells bear the tensional forces generated in the glomerular capillary wall. Not only mesangial cells per se, but also their intercellular junctions should be physically stable against the tensional forces; this prompted the search for actin filament-reinforced adherens junctions of mesangial cells. We previously reported alpha- and beta-catenins localized at the cell-cell contact sites of mesangial cells in the rat. Classical cadherin expressed by mesangial cells, however, remains to be elucidated. METHODS: Expression of classical cadherins, especially N-cadherin, was examined in rat glomeruli by ribonuclease protection assay, Western blot analysis and immunofluorescence and immunoelectron microscopy. RESULTS: Ribonuclease protection assay detected significant expression of N-cadherin in rat glomeruli. Western blot analysis showed that rabbit and murine antibodies against N-cadherin reacted with a specific band in isolated glomeruli. Immunofluorescence microscopy revealed that both antibodies reacted only with the mesangium in glomeruli. Immunoelectron microscopy demonstrated that the immunogold particles for N cadherin were found predominantly at cell-cell contact sites of mesangial cells where actin filaments concentrated. CONCLUSION: N-cadherin interconnects mesangial cells, suggesting that the cadherin-catenin-actin filament system in the mesangium may play a role in the counteraction of the hydraulic pressure gradient across the capillary wall. PMID- 19546591 TI - Mutational analysis of CLC-5, cofilin and CLC-4 in patients with Dent's disease. AB - BACKGROUND/AIMS: Dent's disease is caused by mutations in the chloride/proton antiporter, CLC-5, or oculo-cerebro-renal-syndrome-of-Lowe (OCRL1) genes. METHODS: Eighteen probands with Dent's disease were investigated for mutations in CLC-5 and two of its interacting proteins, CLC-4 and cofilin. Wild-type and mutant CLC-5s were assessed in kidney cells. Urinary calcium excretion following an oral calcium challenge was studied in one family. RESULTS: Seven different CLC 5 mutations consisting of two nonsense mutations (Arg347Stop and Arg718Stop), two missense mutations (Ser244Leu and Arg516Trp), one intron 3 donor splice site mutation, one deletion-insertion (nt930delTCinsA) and an in-frame deletion (523delVal) were identified in 8 patients. In the remaining 10 patients, DNA sequence abnormalities were not detected in the coding regions of CLC-4 or cofilin, and were independently excluded for OCRL1. Patients with CLC-5 mutations were phenotypically similar to those without. The donor splice site CLC-5 mutation resulted in exon 3 skipping. Electrophysiology demonstrated that the 523delVal CLC-5 mutation abolished CLC-5-mediated chloride conductance. Sixty percent of women with the CLC-5 deletion-insertion had nephrolithiasis, although calcium excretion before and after oral calcium challenge was similar to that in unaffected females. CONCLUSIONS: Three novel CLC-5 mutations were identified, and mutations in OCRL1, CLC-4 and cofilin excluded in causing Dent's disease in this patient cohort. PMID- 19546592 TI - Neonatal maternal separation increases brain-derived neurotrophic factor and tyrosine kinase receptor B expression in the descending pain modulatory system. AB - Neonatal maternal separation (NMS) could trigger long-term changes in the central neuronal responses to nociceptive stimuli in rats. Stress-induced visceral hyperalgesia is closely associated with the dysfunction of descending pain modulatory systems. Brain-derived neurotrophic factor (BDNF) not only has an important role in long-term synaptic plasticity but also in facilitating descending pain. The present study aimed to investigate changes in the expression of BDNF and its receptor tyrosine kinase receptor B (TrkB) in the amygdala and the rostral ventromedial medulla (RVM) after NMS and colorectal distention (CRD) stimulation in rats. Male Wistar rat pups were subjected to 180 min of daily NMS or not handled for 13 consecutive days. Expression of BDNF and TrkB following NMS and CRD stimulation was determined using immunohistochemistry. The results revealed an increase in the expression of BDNF and TrkB in the amygdala after NMS. An interactive effect of NMS and CRD on the expression of TrkB, but not BDNF, was found in the RVM. Furthermore, a significant interactive effect of NMS and CRD on the colocalization coefficient of TrkB and phospho-extracellular signal-regulated kinase expression in both the amygdala and RVM were found. These data demonstrate that NMS increases BDNF and TrkB expression in the descending pain systems, which may contribute to the development of NMS-induced visceral hyperalgesia. PMID- 19546594 TI - [Gastrointestinal tumors. Foreward]. PMID- 19546593 TI - Dopaminergic modulation of afferent synaptic transmission in the semicircular canals of frogs. AB - Using multiunit recording of action potentials from the whole nerve with the aid of external perfusion, we investigated the effects of dopamine (DOP) agonists that are involved in modulatory actions on synaptic transmission in the isolated labyrinth preparations of frogs. The external application of DOP (0.1-1 mM), the D(1) agonist chloro-APB hydrobromide (CAPB, 50-100 microM) and the D2 agonist quinerolane (QUI, 50-100 microM) induced a dose-dependent and reversible decline in the resting discharge frequency. In this concentration range, the potency of applied CAPB considerably exceeded that of QUI. AMPA, NMDA and ACPD responses were inhibited by the D1 and D2 agonists, implicating both subtypes of DOP receptors in the modulation of both ionotropic and metabotropic glutamate receptors. The inhibitory action of the DOP agonists on L-glutamate responses persisted in a high Mg2+ solution in conditions of selective activation of the postsynaptic membrane. The results obtained suggest that DOP may interact with both D1 and D2 receptor subtypes, most likely located postsynaptically on the afferent nerve fibers. This dopaminergic control mechanism may result in the reduction of the activated firing rate, thus preventing over-excitation and excitotoxic injury of the afferent dendrites after the external application of L glutamate and excessive receptor stimulation. PMID- 19546595 TI - [Laparoscopic surgery and adjuvant therapy for colon cancer]. AB - At present, about 10% of all oncological procedures in the colon are carried out laparoscopically. Acceptance is increasing. After successful R0 resection, the rule for stage III patients is: adjuvant therapy is indicated regardless of age. Regimens containing oxaliplatin should be used. If there are contraindications for oxaliplatin, then fluoropyrimidine monotherapy is indicated, with oral fluoropyrimidines (capecitabine) being given precedence over infusional schemes. The use of 5-FU bolus regimens is regarded as obsolete. For stage II, the following applies: If an adjuvant chemotherapy is planned in these patients on the basis of the QUASAR data, then fluoropyrimidine monotherapy (e. g. capecitabine) can be given. Since patients whose tumours show a high frequency of microsatellite instability (MSI) do not benefit from a fluoropyrimidine monotherapy, the MSI status should be determined before choosing therapy. PMID- 19546596 TI - [Diagnosis and treatment of liver metastases from primary colorectal tumour]. AB - Contrast-enhanced multislice computer tomography (MSCT) has established itself as the standard tomographic imaging method both for diagnosis and for treatment monitoring of hepatic lesions. To clarify local conditions before partial liver resection, diffusion-weighted magnetic resonance tomography (DWI-MRT) can also provide important additional information. In order to meet the criteria for a R0 resection, a margin of 0.5 mm seems to be sufficient. Neoadjuvant chemotherapy aiming to reduce tumour size can be given in parallel with portal artery embolisation without adversely affecting perioperative morbidity and mortality. As far as the management of primary resectable liver metastases is concerned, there is an urgent need for more studies. Despite the relatively limited evidence, adjuvant chemotherapy is currently more widely favoured in Germany than perioperative chemotherapy. There is also considerable need for studies concerning preoperative therapy in patients with liver metastases that are not (yet) resectable. In KRAS wild-type tumours, high response rates (in terms of a reduction in the size of metastases) are achieved with a cetuximab/chemotherapy combination. Bevacizumab/chemotherapy combinations lead to high rates of pathohistological complete and partial remissions. What the best parameter for judging the success of preoperative therapy is remains unknown, and so comparison studies using survival as a 'hard' endpoint must be carried out. PMID- 19546597 TI - [Palliative treatment for colorectal cancer]. AB - Before a decision is made to give a particular drug treatment, first of all the best strategy for the individual patient must be determined. In a patient with an aggressive tumour, for whom a secondary curative approach by means of metastasis resection is not an option, the preferred first-line treatment will generally be a triple combination therapy containing bevacizumab - and this is also true in KRAS/BRAF wild-type patients, since the main aim here is to achieve the longest possible survival time with a minimum of side effects. If an epidermal growth factor receptor (EGFR) antibody (cetuximab or panitumumab) is to be used in first line or later therapy, then the presence of a KRAS mutation must be excluded beforehand. It is very likely sensible also to exclude a BRAF mutation. Second line treatment after a first-line therapy containing bevacizumab may be a combination chemotherapy or, in patients who are KRAS wild-type (and possibly also BRAF wild-type), irinotecan plus cetuximab. Locoregional treatments such as chemoembolisation, selective internal radiation therapy (SIRT), and stereotactic PMID- 19546598 TI - [Neoadjuvant and surgical treatment for rectal cancer]. AB - According to the 2008 guidelines on colorectal cancer, whether preoperative therapy is indicated for rectal cancer should be judged based on the T and N categories. A few centres limit the indication for preoperative radio(chemo)therapy to patients with tumours that, according to magnetic resonance tomography (MRT), extend to the fascia mesorectalis or are 1 mm or less away from it - so-called circumferential resection margin-positive or CRM positive tumours. Omitting preoperative therapy for MRT CRM-negative tumours is, however, a matter that still requires further study in clinical trials. The high rate of distant metastases continues to be a problem. Assuming that pathohistological complete remission (pCR) is a predictive marker of long-term disease-free survival after neoadjuvant radiochemotherapy, attempts are now being undertaken to intensify the neoadjuvant therapy. Phase II trials show improved pCR rates by combining the preoperative radiation with the double combinations oxaliplatin or irinotecan plus infusional or oral 5-FU (capecitabine). In the case of limited T1 rectal cancer without further risk factors, transanal local excision can be used. PMID- 19546599 TI - Prevention and treatment of corneal neovascularization: comparison of different doses of subconjunctival bevacizumab with corticosteroid in experimental rats. AB - BACKGROUND: To evaluate and compare the effect of different doses of subconjunctival bevacizumab with betamethasone on the development of corneal major new vessels in a rat model of corneal chemical injury. METHODS: The right eyes of 100 male Sprague-Dawley rats were randomly divided into 10 experimental groups (n = 10 per group). Chemical cauterization of the cornea was performed by using silver nitrate/potassium nitrate sticks. Immediately following corneal cauterization, the animals in groups 1-5 received subconjunctival injections of 0.02 ml of normal saline (control A), betamethasone LA (6 mg/ml) and different doses of bevacizumab (1, 5 and 25 mg/ml), respectively. In another experiment, the animals in groups 6-10 received subconjunctival injections of 0.02 ml of normal saline (control B), betamethasone LA (6 mg/ml) and different doses of bevacizumab (1, 5 and 25 mg/ml), respectively, 7 days following corneal cauterization. The numbers of major thick-walled vessels originating from the limbus reaching the corneal scar were counted 7 days after corneal cauterization in groups 1-5 and 14 days after corneal cauterization in groups 6-10. RESULTS: The number of major vessels in groups 1-5 was 19.63 +/- 3.77, 17.25 +/- 5.33, 16.10 +/- 5.02, 12.89 +/- 2.70 and 12.36 +/- 4.45 when assessed 7 days after corneal cauterization, respectively. Administration of betamethasone in group 2 had no significant effect on the corneal major vessel count compared to control A. The number of major vessels in groups 4 and 5 (bevacizumab 5 and 25 mg/ml) was significantly lower than that of group 1 (p < 0.01, Student's t test). The number of vessels in groups 6-10 was 12.55 +/- 5.64, 11.30 +/- 9.33, 5.50 +/- 6.34, 2.73 +/- 4.73 and 2.67 +/- 3.77 when assessed 14 days after corneal cauterization, respectively. Subconjunctival administration of betamethasone 7 days after corneal cauterization did not reduce the amount of corneal major vessels compared to control B. Administration of 0.02 ml of bevacizumab in doses of 1, 5 and 25 mg/ml 7 days after corneal cauterization significantly reduced the amount of major vessels compared to group 6 (p = 0.01, p < 0.01 and p < 0.01, respectively). There was no significant difference in percent area of corneal scar between different groups. CONCLUSION: Single subconjunctival injection of bevacizumab is efficacious in the prevention of formation as well as regression of major vessels compared to betamethasone in this rat model of corneal neovascularization. Even lower doses of bevacizumab might be efficacious. PMID- 19546600 TI - Pentosidine - a new biochemical marker in diabetic retinopathy. AB - AIM: This study was done to evaluate the role of pentosidine in predicting the progression of diabetic retinopathy. PATIENTS AND METHODS: We included 30 subjects with insulin-dependent diabetes mellitus (type 1) and 10 healthy control individuals. A case control study was done. Full ophthalmological examination together with laboratory investigations (blood glucose level, HbA(1C) and blood pentosidine level corrected to blood total protein values were measured) were done in all subjects included in this study. The level of pentosidine was correlated with the duration of diabetes and the stage of retinopathy. All data were analyzed and reported. RESULTS: Significant elevation of pentosidine was found in patients during the earliest detectable phase of diabetic retinopathy (early nonproliferative diabetic retinopathy) and more elevation at the preproliferative stage of retinopathy, returning to lower levels at the proliferative stage of diabetic retinopathy. CONCLUSION: Pentosidine can be used as a biochemical marker for early occurrence of diabetic retinopathy and as an alarming factor in the preproliferative stage of diabetic retinopathy, thus helping decrease ocular complications. PMID- 19546601 TI - Systematic review of intraocular pressure-lowering effects of adjunctive medications added to latanoprost. AB - OBJECTIVE: To evaluate the intraocular pressure (IOP)-lowering effects of adjunctive medications when added to 0.005% latanoprost taken once daily. METHODS: Pertinent publications were identified through systematic searches of PubMed, Embase, and the Cochrane Controlled Trials Register. Randomized clinical trials with over 85% of patients presenting with primary open-angle glaucoma or ocular hypertension who were treated with the combination treatment of latanoprost were selected. The pooled additional IOP-lowering effects at 1-3 months after a run-in phase of at least 2 weeks on 0.005% latanoprost once daily were calculated using the random effects model. RESULTS: Nine randomized clinical trials were included. The mean pooled IOP reductions were 3.3 mm Hg (95% CI: 2.1 4.5) at trough and 4.4 mm Hg (95% CI: 3.4-5.4) at peak when adding 0.5% timolol once daily, 2.6 mm Hg (95% CI: 1.9-3.3) at trough and 3.8 mm Hg (95% CI: 2.5-5.2) at peak when adding 0.1/0.15% brimonidine twice daily, 2.6 mm Hg (95% CI: 1.7 3.4) at trough and 3.1 mm Hg (95% CI: 2.6-3.6) at peak when adding 2% dorzolamide twice daily, 2.4 mm Hg (95% CI: 2.0 -2.8) at trough and 2.7 mm Hg (95% CI: 2.2 3.2) at peak when adding 0.5% timolol twice daily, and 2.8 mm Hg (95% CI: 1.5 4.1) at trough and 1.8 mm Hg (95% CI: 1.2-2.3) at peak when adding 1% brinzolamide twice daily. CONCLUSIONS: The addition of brimonidine, dorzolamide, timolol, or brinzolamide can further lower IOP in eyes being treated with latanoprost. Timolol 0.5% once daily might be the most effective adjunctive medication. PMID- 19546603 TI - 'Keep at it! Accept the challenges of your critics'. An interview with John M. Howard, MD, professor Emeritus, Division of General Surgery, University of Toledo, Toledo, Ohio, USA. Interview by Martin E. Fernandez-Zapico. AB - In this article, Professor John Howard shares with our readers his life experiences as a pancreatic surgeon-researcher and gives advice to junior pancreatologists starting a career in pancreatic research. Professor Howard made an outstanding contribution to the understanding of acute pacreatitis and he is a pioneer in the development of management approaches for pancreatic diseases. PMID- 19546602 TI - Cell-cell adhesion in the prelaminar region of the optic nerve head: a possible target for ionic stress. AB - PURPOSE: This paper aims to study the anterior surface of the optic nerve in relation to its ability to support a source of stress acting from the vitreous cavity. The intercellular junctions of the lining astrocytes mediated by cellular adhesion molecules (CAMs) may be the main targets for ionic stress. METHODS: The optic nerve of the domestic pig was prepared for light, confocal laser and transmission electron microscopy. Immunostaining was performed for antibodies against glial fibrillary acidic protein, neural cadherin (N-cadherin) and neural CAM (N-CAM). RESULTS: Only 1 type of intercellular junction was found among the bordering astrocytes, which was characterized as a zonula adherens. Unions between lining cells showed a positive immunogold effect and immunofluorescence against N-cadherin in the zonula adherens and membrane apposition. N-CAM was also present in areas of nonjunctional cellular adhesion. CONCLUSION: The stability of intercellular junctions of the nerve-vitreous boundary is sensitive to altered concentrations of Ca(2+). Since aqueous humor has half the Ca(2+) concentration of plasma, any contact of aqueous humor with the optic nerve head can interfere with the ionic concentration of calcium in the extracellular spaces. This mechanism may contribute to age-related changes and some types of glaucoma. PMID- 19546604 TI - Tyrosine kinase inhibitors and solid tumours: case report and review of the literature. AB - Bcr-Abl tyrosine kinase inhibitors (TKIs) such as imatinib or dasatinib produce high cytogenetic response rates in patients with Philadelphia-positive chronic myeloid leukaemia (CML) with a good overall safety profile. Despite a complete molecular response, it is currently recommended to continue these targeted therapies to avoid relapse. The immediate and short-term TKI side effects are well known, but the long-term side effects have not yet been clearly identified. A preclinical study in rats treated with TKI showed a statistically significant increase in benign and malignant renal tumours. The authors report the case of a 61-year-old man with CML treated with imatinib with a good response, and they switched to dasatinib after grade 4 hepatic toxicity. He had received treatment with 400 mg of imatinib per day for 77 days, followed by dasatinib for 133 days. He developed a metastatic carcinoma of unknown origin during TKI therapy. Despite chemotherapy, the patient died 2 months after the diagnosis. Although several cases of solid tumours have been reported during TKI therapy, the link between cancer and TKIs is not yet clear. Imatinib has remarkably improved the prognosis of patients with CML. Monitoring of the long-term safety profile of TKIs is essential due to the prolonged survival of these patients. PMID- 19546605 TI - Endothelial dysfunction in the mesenteric artery and disturbed nonadrenergic noncholinergic relaxation of the ileum due to intestinal ischemia-reperfusion can be prevented by sildenafil. AB - The aim of the present study was to evaluate the effects of sildenafil on endothelium-dependent mesenteric artery vasorelaxation and nonadrenergic noncholinergic (NANC) ileal responses in an experimental rat intestinal ischemia reperfusion (I/R) model. The superior mesenteric artery was occluded for 45 min of ischemia and then the clamp was removed for 60 min of reperfusion. Sildenafil (1 mg/kg, i.v.) or saline was administered prior to surgery in the I/R and sham operated groups. Acetylcholine-induced relaxation of the mesenteric arteries, which were precontracted via submaximal phenylephrine, decreased markedly after I/R. Sildenafil pretreatment reversed the acetylcholine-induced relaxation. In the ileum, NANC responses were significantly attenuated following I/R, which were increased by sildenafil pretreatment. These results indicate that pretreatment with sildenafil prevented both endothelial dysfunction in the mesenteric artery and impairment of ileal NANC responses in a rat intestinal I/R model. PMID- 19546606 TI - Over-expression of CDH22 is associated with tumor progression in colorectal cancer. AB - CDH22, a member of the cadherins family, is highly expressed in the pituitary gland and the brain. It has been previously found that CDH22 is involved in morphogenesis and tissue formation in neural and non-neural cells of the brain and neuroendocrine organ. However, little is known about its role in colorectal cancer. Here the role of CDH22 in colorectal tumor was tested, and effects of inhibition of CDH22 expression on proliferation and metastasis were examined. We found that CDH22 was over-expressed at both transcriptional and translational levels in colorectal cancer and lymphatic metastasis of colorectal carcinoma, compared with normal colorectal mucosa. CDH22 over-expression was significantly associated with invasion and metastasis of colorectal cancer both at protein and mRNA levels. CDH22 knockdown partially blocked proliferation of colorectal cancer cells in vivo and in vitro. CDH22 knockdown was sufficient to attenuate invasion of colorectal cancer cells and inhibit tumor metastasis in a mouse model of colon surgical orthotopic implantation. Our results reveal for the first time a new role of CDH22 in progression of colorectal cancer. PMID- 19546607 TI - Update on percutaneous needle biopsy of nonmalignant breast lesions. AB - Certain nonmalignant lesions encountered on percutaneous breast biopsies pose dilemmas with regard to the most appropriate clinical management subsequent to needle biopsy (ie, surgical excision vs. follow-up). These lesions include columnar cell lesions, atypical ductal hyperplasia, lobular neoplasia, papillary lesions, radial scars, fibroepithelial lesions, and mucocele-like lesions. As minimally invasive diagnostic procedures are now standard it is more important than ever to be aware of the limitations of percutaneous biopsy, particularly with regard to apparently benign lesions because of the risk that the radiologically detected lesion may harbor malignant disease not represented in the biopsy specimen. This underscores the importance of radiologic-pathologic correlation. Increasingly, radiologists are adopting vacuum-assisted devices using larger gauge needles. The changing practices among radiologists are reflected in recent studies which have enriched the literature. In addition, magnetic resonance imaging is being used more frequently in breast imaging, resulting in pathologists more often encountering benign biopsies with uncertain imaging correlation. These changes prompted evaluation of the recent literature and its possible effect on management concerns. This review focuses on management issues following the diagnosis of nonmalignant lesions diagnosed on percutaneous breast biopsy and highlights imaging terms commonly used in breast radiology reports to facilitate accurate radiologic-pathologic correlation. PMID- 19546608 TI - KRAS mutation testing in colorectal cancer. AB - In the US, colorectal cancer is the third leading cause of cancer-related death. Approximately 20% of patients present with metastatic disease, and an additional 30% to 40% develop metastasis during the course of their disease. Patients with metastatic colon cancer have a 5-year survival rate of only 11%. Although surgery is the mainstay of treatment for early stage colon cancer, adjuvant treatment is usually used in patients advanced stage disease. In particular, epidermal growth factor receptor (EGFR) inhibitor therapies have emerged as effective treatments in a subset of patients with metastatic colorectal carcinoma. Two anti-EGFR biologics, cetuximab and panitumumab, have been approved by the Food and Drug Administrations for the treatment of refractory metastatic colorectal carcinoma. Mounting evidence has shown that these therapies are ineffective in tumors with mutations of codons 12 and 13 of exon 2 of the KRAS gene. Because of this compelling data, the National Comprehensive Cancer Network and the American Society of Clinical Oncology have recommended determination of KRAS mutation status in all patients with metastatic colorectal cancer who are candidates for anti-EGFR therapy. Anatomic pathologists play an integral role in coordinating the testing for KRAS mutations, as this assay is performed on tissue samples selected by the pathologist. Herein, the authors present an up-to-date review of the biologic, clinical, and laboratory aspects of KRAS mutation testing in colorectal cancer. PMID- 19546609 TI - Multigene classifiers, prognostic factors, and predictors of breast cancer clinical outcome. AB - A series of multigene classifiers, prognostic and predictive tests have recently been introduced as potentially useful adjuncts for the management of recently diagnosed breast cancer patients. These tests have used both slide-based methods including immunohistochemistry and fluorescence in situ hybridization and nonmorphology driven molecular platforms including quantitative multiplex real time polymerase chain reaction and genomic microarray profiling. In this review, a series of partially and completely commercialized multigene assays are compared with the standard breast cancer clinico-pathologic variables and biomarkers and evaluated as to the level of their scientific validation, current clinical utility, regulatory approval status, and estimated cost-benefit. A comparison of the Oncotype Dx and MammaPrint assays indicates that the Oncotype Dx test has the advantages of an earlier commercial launch in the US, wide acceptance for payment by third party payors, the ease of use of formalin fixed paraffin embedded tissues, a recommendation as ready for use by the American Society of Clinical Oncology Breast Cancer Tumor Markers Update Committee, a continuous rather than dichotomous algorithm, inclusion of both estrogen receptor (ER) and human epidermal growth factor receptor 2 in the mRNA profile, an ability to serve as both a prognostic and predictive test for certain hormonal and chemotherapeutic agents, demonstrated cost-effectiveness in 1 published study, and a high accrual rate for the prospective validation clinical trial (Trial Assigning Individualized Options for Treatment Rx). The MammaPrint assay has the advantages of a 510(k) clearance by the US Food and Drug Administration, a larger gene number which may enhance further utility, and the potentially wider patient eligibility including lymph node-positive, ER-negative, and younger patients being accrued into the prospective trial (the Microarray in Node-negative Disease may Avoid ChemoTherapy). A number of other assays have specific predictive goals most often focused on the efficacy of tamoxifen in ER-positive patients such as the Two-gene Ratio test and the Cytochrome P450 CYP2D6 genotyping assay. PMID- 19546610 TI - Diffuse aggressive B-cell lymphomas. AB - Diffuse aggressive B-cell lymphomas comprise a relatively common and increasingly diverse group of neoplasms. Newer modalities including gene expression profiling and an increasing panel of immunohistochemical markers have contributed to greater accuracy in defining these entities. Attention is paid not only to the neoplastic cells but also to the cellular and stromal milieu in which they proliferate. These distinctions may have therapeutic implications as well, with improved outcome related to newer and sometimes targeted therapies. At the same time there is increasing understanding of the overlap, which occurs in the grey zone between diffuse large B-cell lymphoma and Burkitt lymphoma as well as between diffuse large B-cell lymphoma and Hodgkin lymphoma. This review aims to provide practical insights in the correct identification and differential diagnosis of these lymphomas, with emphasis on the changes that have occurred with the publication of the 2008 World Health Organization updated classification. PMID- 19546611 TI - Castleman disease: an update on classification and the spectrum of associated lesions. AB - Since its initial description, researchers have expanded the spectrum of Castleman disease to include not only the classic and well-recognized hyaline vascular type, but also the plasma cell type and multicentric types of broader histologic range, including human herpes virus-8-associated Castleman disease. These less common subtypes of Castleman disease are less familiar, and may be under-recognized. Also of practical importance, current authors are restructuring the classification of multicentric Castleman disease to accommodate the emerging pathogenic role of human herpes virus-8 and its association with the recently described plasmablastic variant. In addition to an increased risk of lymphoma, patients with Castleman disease also are at increased risk for other related neoplasms, including Kaposi sarcoma and follicular dendritic cell tumors, which are of prognostic and therapeutic relevance. This review focuses on the histologic diagnosis of Castleman disease, current and emerging concepts in its pathogenesis and classification, and associated histopathologic entities. PMID- 19546612 TI - Goblet or signet ring cells: that is the question. AB - Goblet cell carcinoid tumor is a rare mixed endocrine-exocrine neoplasm of the appendix. It carries an intermediate biologic behavior between a classic carcinoid tumor and a conventional adenocarcinoma. There has been a general lack of clinicopathologic parameters that can be reliably used to predict the clinical course and patient outcome. A recent retrospective study of a large number of appendiceal goblet cell carcinoids has shown that these tumors can be stratified into 3 subgroups based on careful histologic analysis: typical goblet cell carcinoid (group A); adenocarcinoma ex goblet cell carcinoid, signet ring cell type (group B); and adenocarcinoma ex goblet cell carcinoid, poorly differentiated carcinoma type (group C). Clinical follow-up data show that the histologic classification correlates with patient survival. Thus, meticulous histologic evaluation is of critical importance in determining the prognosis and guiding the management of patients with goblet cell carcinoids. This brief review summarizes these recent findings and raises a few issues that may need to be further addressed to better apply this classification to our practice. PMID- 19546613 TI - Fatal systemic venous air embolism during endoscopic retrograde cholangiopancreatography. AB - Hepatic portal venous air embolism is the rarest complication of gastrointestinal endoscopy, resulting from penetration of gas into the portal veins, and may occur during endoscopic retrograde cholangiopancreatography and endoscopic biliary sphincterotomy. The likely mechanism is intramural dissection of insufflated air into the portal venous system through duodenal vein radicles transected during the procedure. Hepatic portal air embolism may be fatal. Cerebral air embolism may also occur. So far 13 cases of air embolism after endoscopic retrograde cholangiopancreatography have been reported, with 4 cases of systemic spread that proved fatal. Death was due to pulmonary air embolism in 2 cases, and cerebral air embolism in another 2. We report on an additional such fatal case, concerning a 78-year-old male patient, who several years previously had undergone surgical gastroduodenal resection with cholecystectomy and papillotomy, and was admitted for recurrent ascending cholangitis secondary to bile duct stones. During the third endoscopic cholangioscopic procedure for removal of bile duct stones, sudden cardiopulmonary arrest occurred. Death was due to massive pulmonary air embolism. Cerebral air embolism was also found. Autopsy was performed. A spontaneous duodenobiliary fistula was found. On the basis of bench radiologic investigation (retrograde suprahepatic venography and anterograde portography), it was demonstrated that the air insufflated during duodenal endoscopy, which entered through the spontaneous duodeno-biliary fistula, penetrated into intrahepatic vein radicles injured secondarily to prolonged impaction of biliary sand and stones and infection, resulting in portal and hepatic venous gas and systemic air embolism. PMID- 19546617 TI - Postadoption depression: what nurses should know. Adoptive parents need support, too. PMID- 19546618 TI - More on the national nurse. PMID- 19546619 TI - Medevac risks. PMID- 19546623 TI - Nursing bra brouhaha. PMID- 19546624 TI - Nursing bra brouhaha. PMID- 19546626 TI - Who's Watching Nurses? PMID- 19546635 TI - HIPAA: not so bad after all? PMID- 19546636 TI - Influenza A (H1N1): a wake-up call. PMID- 19546637 TI - Street nursing. AB - A British Columbia outreach program aims at preventing sexually transmitted infections and HIV. PMID- 19546639 TI - 'It depends': medical residents' perspectives on working with nurses. AB - OBJECTIVE: Using the theory of relational coordination, which holds that in high pressure settings such as hospitals, high-quality communication and strong relationships are necessary for coordinated action, we sought to determine the quality of the nurse-physician relationship by examining the communication and interaction between nurses and residents from the residents' perspective. METHODS: A sample of 20 medical and surgical residents, selected by a snowball sampling technique, were interviewed about the quality of their communication and relationships with nurses in the workplace. RESULTS: Residents' responses were influenced by their perceptions of nurses' cooperativeness and competence, and their impressions of nurses' professional preparation and demeanor varied widely. Although 19 of 20 residents reported instances of poor communication or problematic relationships with nurses, most believed that this posed no significant threat to patient care because the nurses' role, as they saw it, was one of simply following orders. CONCLUSIONS: Given the strong doubts some residents expressed about nurses' cooperativeness and competence, the nursing profession should consider strengthening nursing education and clearly delineating nurses' roles and competencies. PMID- 19546644 TI - Post-breast cancer. Lymphedema: part 1. AB - Lymphedema, which can be a debilitating sequela to breast cancer treatment, is characterized by an abnormal accumulation of lymph in the arm, shoulder, breast, or thoracic area. It may appear gradually or suddenly, and although it usually develops within three years of a breast cancer diagnosis, it can arise much later; survivors remain at lifetime risk. The condition can cause physical discomfort and pain, impaired function, and emotional distress. It's imperative that survivors' risk of lymphedema be reduced and that those who develop it receive help to manage it. Part 1 of this two-part article describes post-breast cancer lymphedema and discusses its diagnosis and measurement. Part 2 (next month) will discuss risk reduction, treatment, and implications for nurses. PMID- 19546646 TI - Men's Health Pitstop. AB - A nurse-coordinated program to bring health screening to rural farmers. PMID- 19546647 TI - How neighborhood environments contribute to obesity. PMID- 19546649 TI - Safe and reliable care. AB - This is the 11th in a series of articles from Massachusetts General Hospital describing one general medical unit's experiences with Transforming Care at the Bedside (TCAB). An initiative begun by the Robert Wood Johnson Foundation (RWJF) and the Institute for Healthcare Improvement, TCAB was developed to improve care on medical-surgical units, patients' and family members' experience of care, and teamwork among care team members and to increase nurse satisfaction and retention. The TCAB philosophy empowers bedside nurses to generate ideas and solutions for change. Mass General is one of 68 hospitals participating in a two year TCAB initiative led by the American Organization of Nurse Executives and funded with a grant from the RWJF. For more information on TCAB, go to www.rwjf.org/pr/product.jsp?id=31512. PMID- 19546648 TI - Conflict in the workplace: part 2. AB - Last month, in Part 1 of this two-part article, I explored factors that contribute to workplace conflict among nurses (such as sex, age, power, and culture), as well as individual responses to conflict. I also discussed my observation that nurses apply their skills in therapeutic communication to solving workplace conflict, and that they therefore tend to focus on emotions rather than on solutions. In Part 2, I present strategies nurses can use to resolve conflict and build more effective-and harmonious-workplace relationships. PMID- 19546650 TI - Intake interview. Gender: it's not an emergency, is it? PMID- 19546652 TI - New research in low-tech solutions to a significant public health problem: assessment of depression in the elderly. PMID- 19546653 TI - Alcohol consumption as a risk factor for dementia and cognitive decline: meta analysis of prospective studies. AB - The relationships between alcohol consumption and dementia and cognitive decline were investigated in a systematic review including meta-analyses of 15 prospective studies. Follow-ups ranged from 2 to 8 years. Meta-analyses were conducted on samples including 14,646 participants evaluated for Alzheimer disease (AD), 10,225 participants evaluated for vascular dementia (VaD), and 11,875 followed for any type of dementia (Any dementia). The pooled relative risks (RRs) of AD, VaD, and Any dementia for light to moderate drinkers compared with nondrinkers were 0.72 (95% CI = 0.61-0.86), 0.75 (95% CI = 0.57-0.98), and 0.74 (95% CI = 0.61-0.91), respectively. When the more generally classified "drinkers," were compared with "nondrinkers," they had a reduced risk of AD (RR = 0.66, 95% CI = 0.47-0.94) and Any dementia (RR = 0.53, 95% CI = 0.53-0.82) but not cognitive decline. There were not enough data to examine VaD risk among "drinkers." Those classified as heavy drinkers did not have an increased risk of Any dementia compared with nondrinkers, but this may reflect sampling bias. Our results suggest that alcohol drinkers in late life have reduced risk of dementia. It is unclear whether this reflects selection effects in cohort studies commencing in late life, a protective effect of alcohol consumption throughout adulthood, or a specific benefit of alcohol in late life. PMID- 19546654 TI - Clinically significant depressive symptoms and associated factors in community elderly subjects from Sao Paulo, Brazil. AB - OBJECTIVES: To determine the frequency of clinically significant depressive symptoms (CSDS) in a community sample of Brazilian elderly and to assess their relationship with sociodemographic factors, cognitive and functional impairment (CFI), and clinical diseases. DESIGN: Cross-sectional study of a community-based sample of elderly subjects. SETTING: City of Sao Paulo, State of Sao Paulo, Brazil. PARTICIPANTS: A total of 1,563 elderly subjects aged 60 years or older. MEASUREMENTS: A 10-item scale for screening of depressive symptoms in elderly people (D-10), the Mini Mental State Examination, the Fuld Object Memory Evaluation, the Informant Questionnaire on Cognitive Decline in the Elderly, the Bayer Activities of Daily Living Scale, and a sociodemographic and clinical questionnaire. RESULTS: The frequency of CSDS was 13.0%. Univariate analysis identified independent factors associated with these symptoms in our sample. Logistic regression analysis indicated that being female, brown skinned, previously depressed, having CFI, using psychotropics, and not practicing physical exercise were related to CSDS. On the other hand, being older, clinically sick, employed, or married were not associated with CSDS. CONCLUSIONS: : Consistent with previous reports, female gender, lack of physical activity, and CFI were significantly associated with higher frequencies of CSDS. Further investigations are necessary to clarify the occurrence of depression and possible modifiable factors in developing countries such as Brazil. PMID- 19546655 TI - Diffusion tensor imaging of the posterior cingulate is a useful biomarker of mild cognitive impairment. AB - OBJECTIVES: Mild cognitive impairment (MCI) is recognized as a predementia state, but its definition is inconsistent and only 20%-30% develop dementia after 2 years. Biomarkers may help identify individuals at greatest risk of progressive decline. The authors examine a novel neuroimaging technique, diffusion tensor imaging (DTI) as a potential biomarker of MCI. DESIGN: Cross-sectional prospective study. SETTING: Subjects were recruited randomly using the electoral roll from two electorates in East Sydney, Australia. PARTICIPANTS: A community dwelling sample (N = 249) and age 70-90 years. MEASUREMENTS: Screening to exclude dementia, comprehensive neuropsychiatric assessment, cognitive test battery, structural magnetic resonance imaging and DTI to obtain measures of fractional anisotropy (FA) and mean diffusivity (MD). MCI was diagnosed by standard criteria. RESULTS: After controlling for age, sex, and years of education, the amnestic MCI (aMCI) group demonstrated microstructural pathology in the parahippocampal white matter, frontal white matter, splenium of corpus callosum, and posterior cingulate region. The nonamnestic MCI (naMCI) group demonstrated microstructural pathology in the frontal white matter, internal capsule, occipital white matter, and the posterior cingulate region. A binary logistic regression model showed that DTI of the left posterior cingulate was significant in identifying persons with aMCI to an accuracy of 85.1%. Receiver operating characteristics curve analysis yielded a sensitivity of 80% and specificity of 60.3% in distinguishing aMCI from naMCI and the normal comparison group. CONCLUSION: DTI of the posterior cingulate region discriminates MCI from cognitively normal individuals with accuracy and has the potential to be used as a biomarker of MCI, in particular aMCI. PMID- 19546656 TI - The association between dementia and long-term use of benzodiazepine in the elderly: nested case-control study using claims data. AB - OBJECTIVES: The aim of this study was to examine the association between long term benzodiazepines (BZDs) use and the risk of dementia. DESIGN: Population based nested case-control study of dementia. SETTING: All subjects were aged 45 and older and enrolled in the National Health Insurance Research Database in Taiwan, 1997-2004. PARTICIPANTS: Cases (N = 779) were patients who were identified with dementia at least two times in their outpatient claims. They were individually matched to six comparison subjects (N = 4,626) based on age and gender. MEASUREMENTS: BZD usage (average dosage per year, average days per year, and cumulative dose and periods) and potential confounding comobidities, including cardiovascular and psychiatric diseases. RESULTS: Subjects with dementia had higher cumulative dose, longer duration of BZDs exposure, and more likelihood to be long-term BZDs users. CONCLUSION: Our findings suggest that long term use of BZDs is associated with an increased risk for dementia, but the underlying mechanisms remain unclear, and further investigations are needed. Long term use of BZDs should be avoided among the elderly, who may be at a higher risk for developing dementia, in addition to other health problems. PMID- 19546657 TI - Changes in vitamin E prescribing for Alzheimer patients. AB - OBJECTIVE: At the end of 2006, a survey was sent to members of the American Association of Geriatric Psychiatry (AAGP) to assess possible changes in prescribing Vitamin E to patients with Alzheimer disease that followed two published reports in early 2005 suggesting increased mortality and an increased incidence of heart failure with Vitamin E supplements. METHOD: A three-item questionnaire was mailed to all AAGP members who had prescription privileges to assess changes in prescribing Vitamin E after January, 2005. RESULTS: A total of 572 completed surveys were returned for a response rate of 35%. Nearly 60% of respondents reported a change over the 2 years that followed the 2005 reports. The greatest change was in the group not prescribing Vitamin E, which increased from 6.6% before 2005 to 60.6% afterward. CONCLUSIONS: AAGP members significantly reduced prescribing Vitamin E to patients with Alzheimer disease after 2005. The two reports are discussed with an emphasis on their methodological limitations and the potential for additional information on Vitamin E side effects from ongoing research. PMID- 19546659 TI - Lower serum levels of lithium may produce efficacy without adverse effects. PMID- 19546660 TI - Surgical gloves and the Food and Drug Administration. PMID- 19546661 TI - Another Barak from earlier times: elucidating the origins of rhinoplasty and solving the "B.L." mystery. PMID- 19546664 TI - Patient draping and endotracheal tube positioning during facelift surgery. AB - A comprehensive approach to facial rejuvenation often requires facelift surgery combined with ancillary facial procedures. This may require prolonged operating time under general anesthesia or conscious sedation. When general anesthesia is used, secure endotracheal tube fixation and ready access to the face is essential. We describe an anesthetic technique that assures secure tube placement, rapid intubation, and ready access to the entire face. A fiber reinforced tube is placed orotracheally, wired to the mandibular dentition, and brought over the head in the fashion similar to a nasotracheal tube. The reinforced nature of the tube prevents kinking and allows rapid repositioning inferiorly to allow access to the upper face. In over 400 rhytidectomies, this technique has allowed rapid intubation and surgical preparation. There have been no airway-related problems or other related complications. This is a safe, effective, and rapid means of securing orotracheal intubation during facial esthetic surgery allowing ready access to the face. PMID- 19546665 TI - Psychological characteristics of Danish women with cosmetic breast implants. AB - An excess of suicide among women with cosmetic breast implants compared with controls has consistently been reported in epidemiologic studies. We have evaluated psychological characteristics among 423 Danish women with cosmetic breast implants, compared with 414 controls. Odds ratios (OR) with 95% confidence intervals (CI) for self-reported psychological symptoms were calculated using multiple logistic regression. Substantial excesses of all studied symptoms before implant surgery were reported among women with breast implants compared with women with other cosmetic surgery, whereas ORs for virtually all symptoms occurring after surgery were close to or below 1.0. In particular, ORs for treatment for depression, cognitive/depressive symptoms, and depression/low spirit before surgery were 4.6 (95% CI = 2.1-10.0), 3.9 (95% CI = 1.9-7.8), and 2.5 (95% CI = 1.1-5.5), respectively. In contrast, the corresponding ORs for these 3 psychological symptoms after surgery were 0.9 (95% CI = 0.6-1.4), 1.0 (95% CI = 0.7-1.5), and 1.0 (95% CI = 0.6-1.5), respectively. In conclusion, women with cosmetic breast implants reported preoperative psychological symptoms indicative of depressive disorders substantially more frequently than women with other cosmetic surgery. Future studies using standardized, validated psychiatric assessment tools are needed to determine whether this can explain the higher risk for suicide among a subset of women seeking cosmetic breast implants. PMID- 19546662 TI - Asia mentor redux. PMID- 19546666 TI - "Scarless" umbilicoplasty: a new umbilicoplasty technique and a review of the English language literature. AB - Many techniques have been described for umbilicoplasty after abdominoplasty, but none of these techniques seems ideal. In this report, we wish to report a new "scarless" umbilicoplasty technique, which bears many of the characteristics of an ideal technique: it is easy to perform and results in the complete absence of visible scars and with a preferred vertical orientation. The aesthetic results of this technique are subjectively and objectively evaluated as compared with the classic umbilicoplasty and these results are discussed among the English language literature. In the period of 2004 to 2005, a series of 138 female patients have had an abdominoplasty with either the classic umbilicoplasty (n = 31) or with our scarless umbilicoplasty (n = 107). After a follow-up period of at least 3 (of 6) months, a questionnaire was send to all of these patients to evaluate patient satisfaction. Twenty-five patients from the classic umbilicoplasty group responded, 53 patients from the scarless umbilicoplasty group. Age (mean 45 with range, 22-66 years) and body mass index (29 with range, 22-35) did not differ among both groups. Also a random selection of fifteen photos from both groups was analyzed and rated according to the system of Strasser by an independent panel. There were no major complications in both groups, but in the classic group, there were some cases with hypertrophic scarring. Patients who underwent the scarless umbilicoplasty technique graded the appearance of their umbilicus significantly better on shape, depth, hygiene, and scar. No significant differences were found in grading size and wound healing. Objective evaluation of the photos demonstrated significant better results for the scarless umbilicoplasty technique. Based on our subjective and objective analysis we conclude that our new technique of the scarless umbilicoplasty features many of the characteristics of the ideal umbilicoplasty: a rather simple and reliable method for creating a natural looking umbilicus when performing an abdominoplasty. PMID- 19546668 TI - Mastopexy techniques after massive weight loss: an algorithmic approach and review of the literature. AB - Mastopexy after massive weight loss (MWL) poses unique challenges that may not be successfully addressed with traditional mastopexy procedures. Several novel techniques have been proposed to improve esthetic outcomes; however, little data exists to guide the plastic surgeon on choice of technique for individual patients. A literature review revealed 10 articles with specific emphasis on mastopexy techniques in MWL patients. These articles focused on ways to improve shape, projection, and long-term results, using autologous tissue alone or combined with breast implants. Key concepts include increasing volume of the breast by utilizing excess axillary tissue (lateral thoracic/spiral/intercostal artery perforator flap), modification of existing superomedial pedicle techniques to maximize breast volume, and increasing breast parenchymal support with suture fixation and dermal suspension. This article offers an algorithmic approach to treat breast ptosis in the MWL patient based on breast volume, axillary tissue, desired scar location, and preferred surgical technique. PMID- 19546667 TI - One hundred forty-one consecutive attempts at autologous tissue single-stage breast cancer reconstruction. AB - The analysis of a single surgeon's 5 year attempt at performing autologous tissue breast cancer reconstruction, with one general anesthetic. A single-stage breast cancer reconstruction is successful if after the original reconstruction, no correction for complications, revision of breast mound, or contralateral breast procedures are performed, under general anesthetic, to complete the reconstruction. This is a review of a single surgeon's breast reconstruction practice. Three hundred fifty-six breast cancer reconstruction patients had surgery in a period of 5 years. One hundred forty-one of 356 (39.6%) were consecutive attempts at single-stage autologous tissue reconstruction: 106 of 141 (75.1%) were free abdominal flaps (transverse rectus abdominis myocutaneous, muscle-sparing flaps [MS], or deep inferior epigastric artery perforator flaps), 29 of 141 (20.6%) pedicled transverse rectus abdominis myocutaneous flaps, and 6 of 141 (4.3%) latissimus dorsi flaps; 37 of 141 (26.2%) were immediate reconstructions, 100 of 141 (70.9%) delayed reconstructions, and 4 of 141 (2.8%) mixed reconstructions. One hundred seven of the 141 patients (75.9%) had their autologous tissue reconstruction successfully performed in one general anesthetic. Reconstructions requiring more than one general anesthetic were due to 18 of 141 (12.8%) postoperative and donor-site complications, 16 of 141 (11.3%) revisions of breast mound or contralateral breast procedures. A total of 34 of 141 (24.1%) reconstructions required a second general anesthetic for successful completion, only 16 of 141 (11.3%) of autologous tissue breast cancer reconstructions required revisions for symmetry. Therefore, single-stage breast cancer reconstruction is feasible and should be attempted to decrease the morbidity of breast cancer survivors. PMID- 19546669 TI - Venous thromboembolic disease in autogenous breast reconstruction. AB - Venous thromboembolic disease (VTE) is a cause of significant morbidity and mortality in patients with cancer. Large studies have estimated that VTE occurs in up to 1.1% of patients undergoing breast cancer tumor extirpation and in up to 1.5% of patients undergoing breast cancer reconstruction. This study sought to retrospectively review the experience of a large university practice with TRAM, DIEP, and latissimus flap reconstruction for mastectomy defects and evaluate our rate of VTE. In our series of 271 consecutive patients, 2 had deep venous thromboses, 2 had both deep venous thromboses and pulmonary emboli, and 2 had pulmonary embolus alone. VTE incidence was 2.2%, a relatively high rate compared with previously published, large population studies of VTE in breast reconstruction patients. Review of the literature suggests that physicians have poor compliance with established guidelines for prophylaxis and treatment of VTE in general and orthopedic surgery populations. Unfortunately, no specific guidelines are available for patients undergoing operative intervention for breast cancer or autogenous tissue based reconstruction. VTE is significantly under-diagnosed: clinical findings alone are unreliable, and the true prevalence may be greater than twice what is reported. Further research is needed in this largely unexplored field to determine appropriate means of VTE prophylaxis and treatment in the breast cancer population. PMID- 19546670 TI - Contralateral internal mammary silicone lymphadenopathy imitates breast cancer metastasis. AB - This case report presents a unique, late complication of breast reconstruction surgery. A woman, who underwent left mastectomy and several reconstruction procedures with silicone implants presented with symptomatic enlarged internal mammary lymph nodes on her contralateral side. The nodes, which were suspicious for breast cancer metastasis on positron-emission tomographic computed tomography, were removed by thoracoscopy. The histopathologic result revealed silicone adenopathy. This report is particularly interesting because it presents a rare case in which silicone has migrated to the contralateral internal mammary nodes. This complication was not previously documented in the medical literature and serves as a possible differential diagnosis to metastatic breast cancer. PMID- 19546671 TI - A new sclerotherapy technique for the wrist ganglion: transcutaneous electrocauterization. AB - Ganglion, a cystic benign mass, most common soft tissue tumor of the hand, usually occurs in hand, wrist, and foot. In this study, we discuss a new sclerotherapy technique through which 17 patients with wrist ganglion were treated by using short bursts of high-frequency low voltage electrodessication delivered through a fine electrode that was inserted into the sac. Their ages varied from 28 to 52 with an average of 32.7 years. Two patients had volar wrist and 15 others had dorsal ganglia. In all patients, an ultrasound imaging was done for the discrimination of the other hand tumors. Under aseptic conditions, first ganglion was aspirated by using a large needle, which was commonly used for peripheric venous catheterization, and 0.5 mL of 1% xylocaine was injected into the cystic cavity, then electrocauterization was done. In the postoperative follow-up ranging from 6 to 29 months, 1 recurrence developed 3 months after the intervention, requiring the same procedure to overcome it. No complication occurred and all complaints of the patients resolved with this approach. The present technique is simple, safe, effective, and inexpensive for ganglion sclerotherapy, resulting in hopeful outcomes to become as an acceptable alternative to the open surgery. PMID- 19546672 TI - The role of freestyle perforator-based pedicled flaps in reconstruction of delayed traumatic defects. AB - The use of perforator-based flaps as freestyle pedicled flaps for traumatic defects has been limited. We explored this possible application in small to moderate sized traumatic defects presenting in the delayed phase, with distinct oedema and induration in the potential flap donor area and posttraumatic vessel disease. Attempts to skeletonize perforator vessels are likely to compromise the flap perfusion, and inadequate dissection is likely to limit mobility of the indurated tissues in the flap. Conventionally, an axial pattern pedicled or a free flap would be needed in such cases, thus increasing its magnitude. We used the freestyle technique to cover traumatic defects by retrograde dissection of pedicled perforator-based flaps. As the surgery was performed in the delayed phase, the tissues were indurated and a larger tissue cuff was preserved around the pedicle than would be our practice in elective surgery. In addition, flap dimensions were planned larger than the defect to be closed. The donor defect was either skin grafted or closed primarily. Our study included 11 cases at various sites over the body. All flaps survived, though 3 flaps encountered major complications, 2 of which needed reoperation. None of the flaps failed completely. The pedicled perforator-based flap provides the surgeon with additional reconstructive options in the setting of trauma. These flaps can be safely harvested using indurated tissue; thus in selected cases, a free flap can be avoided, and reliable cover can be provided with a pedicled flap. Nevertheless, clinical judgment is essential to assess the potential vascular territory of the flap. PMID- 19546673 TI - Vascularized iliac osteomuscular free flaps for composite soft tissue and bone defects of the lower extremity. AB - The purpose of this study to evaluate if vascularized iliac osteomuscular free flaps, which use simple iliac graft companied by covering the iliacus muscle in the inner cortex, is a better option for soft tissue and bone defects of the lower extremity. A retrospective review was performed on 7 consecutive patients treated with vascularized iliac osteomuscular free flaps from June 1999 to March 2003. Survival of skin graft transferred onto the surface of the iliacus muscles provided indirect proof for the survival of transferred bone flap. Skin grafts survived completely in 4 cases and partially in 3 cases. The patients were followed up for 8 to 26 months (average, 14 months) and had acceptable contour and function. PMID- 19546675 TI - Influence of nasal bone fracture and its reduction on the airway. AB - Studies regarding nasal bone fractures have mainly focused on esthetic aspects, but the nose also plays an important role as an airway. The purpose of this study was to evaluate the influence of nasal bone fractures and the treatment of nasal bone fractures on nasal obstruction. Nasal bone fractures were classified by fracture site and septal displacement. Nasal airways were investigated at 3 time points: before reduction (n = 84), 6 days postreduction (n = 84), and 1-year postreduction (n = 27). Airway changes were based on minimal cross-sectional area measurements. The severity of airway obstruction following fracture differed according to the fracture site and the presence of septal displacement. Bilateral fractures caused more profound airway obstruction than unilateral or tip fractures and septal displacement was the most important parameter in nasal obstruction. Airway obstruction improved immediately after reduction by 21% and deteriorated by 4% 1-year postreduction. PMID- 19546674 TI - Bilateral extraoral, infraorbital nerve block for postoperative pain relief after cleft lip repair in pediatric patients: a randomized, double-blind controlled study. AB - The objective of this study was to evaluate the effectiveness of bilateral extraoral infraorbital nerve block with 0.25% bupivacaine administered at the end of surgery in postoperative pain relief after cleft lip repair. Forty ASA I-II children were randomly divided into 2 groups. Group I received 1.5 mL 0.25% bupivacaine and group II received 1.5 mL saline. FLACC scores of the patients in the recovery room in group I were 4 times less than in group II (P = 0.001) and in the first 4 hours postoperatively were apparently less in group I (P = 0.001). Mean time to first paracetamol requirement was longer in group I (P = 0.001). Total paracetamol consumption was lower in group I (P = 0.001). None of the patients required rescue tramadol in group I, whereas all patients in group II needed. In group I, parent satisfaction scores were higher (P = 0.001). Vomiting incidence was higher in group II (P = 0.028). Bilateral extraoral, infraorbital nerve block administered at the end of surgery provides satisfactory analgesia with high parental satisfaction and lower complication rates and reduces rescue analgesic consumption in patients undergoing repair of cleft lip. PMID- 19546676 TI - Galea-including forehead flap for lower one-third nasal reconstruction. AB - The first choice for internal mucosal restoration of the nose is a septal mucosal or vestibular local flap. The forehead flap, raised including the galeal layer, is an alternative option for large nasal defects. It can be used in any difficult situation in which septal or vestibular flaps are not adoptable, such as complete loss of lower one-third. The authors intend to describe the inclusion of galea in the traditional median forehead flap for nasal lining reconstruction. Thirteen patients treated with a forehead flap including galea for lower one-third nasal reconstruction were retrospectively reviewed. No complete flaps necrosis occurred. In 1 case, lining was lost due to infection. In 2 cases a moderate nostril stenosis was observed as late complication. The forehead flap with galea is a good option for large nasal full-thickness defects, involving the lower one third. PMID- 19546677 TI - An intermaxillary fixation screw traction wire: an aid for facial bone fracture repair. AB - We have devised a new technique to improve stabilization of fractured facial bone fractures (frontal sinus fractures, zygomatic fractures, mandibular condyle fractures) by intermaxillary fixation screw traction wires (stainless steel wires through intermaxillary fixation screws). A retrospective study evaluating intermaxillary fixation screw traction wires was performed. We have used this technique for 3 cases of frontal sinus fractures, 9 cases of zygomatic fractures, and 7 cases of mandibular condyle fractures. After dissection of a fractured site, a hole is drilled on the fractured bone where it does not interfere with positioning the plate across the fracture line. After an intermaxillary fixation screw is inserted, a stainless steel wire is tied through a hole in the screw head. By the aid of wire for traction, the displaced fractured bone is easily aligned to the proper position. Plates and screws are applied readily on the predetermined area. A retrospective study on 19 patients using intermaxillary fixation screw traction wires was performed. The diagnoses and associated complications of the cases were recorded. No associated complication as a result of using this technique was identified. The use of intermaxillary fixation screw traction wire enhances stabilization and visualization without possible risk for surrounding soft tissue injury using, a sharp traction device like a bone hook. An intermaxillary fixation screw traction wire is an useful aid for visualization and stabilization during facial bone fracture reduction, particularly where exposure is difficult such as in the condylar region of the mandible. And unlike a classic traction wire, the intermaxillary fixation screw traction wire has almost no risk of having it loosened from the screw. PMID- 19546678 TI - Descending necrotizing soft tissue infection of periodontal origin. AB - Descending necrotizing soft tissue infections involving the neck and chest are a rare but hazardous sequela of periodontal infections. These infections are associated with significant mortality.The high mortality rates of this condition are attributed to significant delays in diagnosis and treatment of common periodontal infections that have progressed to severe disease without overt clinical signs or symptoms. As with all necrotizing soft tissue infections, making a timely diagnosis requires that the examining physician maintain a reasonable index of suspicion when the patient with an oropharyngeal infection is encountered. Coverage of the most common infecting organisms with appropriate antibiotics accompanies surgical therapy. Hyperbaric oxygen remains an adjunctive therapy of unproven efficacy. The cornerstone of therapy, however, is surgical with aggressive and complete debridement of all nonviable tissue without regard for the size or complexity of the resulting wound. In the case described below, this goal could not have been achieved without the immediate availability of reconstructive expertise to preserve vital structures in the neck and upper mediastinum. PMID- 19546679 TI - Modified distally based sural nerve flaps in acute traumatic forefeet reconstructions. AB - Conventional reversed sural flaps have been used to cover lower one-third of the leg defects. However, the experience of the authors indicates that when the soft tissue defect located at the dorsum of the metatarsophalangeal joint, distal marginal necrosis of the flaps usually occurs, which is the exact part of the flap that one needs the most. Finding a new method to augment the blood supply of the flap can be a difficult task. The authors found there is a constant cutaneous branch emanate from the peroneal artery at the point 11.0 +/- 1.7-cm upon the lateral malleolus. Ten modified distally based reverse sural artery flaps, in which the cutaneous branches from the peroneal artery 11.0 +/- 1.7-cm upon the lateral malleolus were added, were performed for the distal-third of the foot reconstruction between 2003 and 2006. All of the flaps survived completely after the operation. Distal marginal necrosis did not occur in any of the flaps. When conventional local flaps are inadequate, this flap should be considered for its reliability and low associated morbidity. PMID- 19546680 TI - Distally based saphenous nerve-great saphenous veno-fasciocutaneous compound flap with nutrient vessels: microdissection and clinical application. AB - Improvements were made by us in several distally based pedicled flaps of the nutrient vessels of the saphenous nerve with lower rotation points. However, these flaps are still insufficient for trauma complicated by bone defects. Accordingly, we conducted a systematic study of the anatomic theory on distally based pedicled compound flaps of the nutrient vessels of the saphenous nerve and great saphenous vein with 30 lower limbs of adult cadavers injected with red gelatin through the femoral artery. It is found that the nutrient vessels of the saphenous nerve-great saphenous vein consist of arteria saphena, fascial cutaneous branches of the inferior medial genicular artery intermuscular spatium branches of the posterior tibial artery, osteocutaneous perforators, superior ankle perforators, medial anterior malleolus perforators, and fascial perforators of the ankle tunnel region. Musculocutaneous perforators of the interior gastrocnemius muscle also enter the nutrient vessels of cutaneous nerve superficial vein. From May 2004 to February 2007, 23 cases of skin flaps for treating defective and infectious wound, 10 cases of musculocutaneous flaps for treating ulcus in the lower segment of the leg, medullitis, and exposure of bone, 3 cases for medial calcaneus medullitis, 7 cases of skeletal flaps for treating tibial defects and nonunion of calcaneal bone. In 2-15-month follow-ups all cases presented with survived flaps, and healed surfaces of the wound and the osteomyelitis. For cases of bone nonunion, it showed that the nonunion healed after 18 weeks, with recovery of work ability after year. Three types of the distally based pedicled flaps or compound flaps of vessels of different perforating branches can be designed for repairing tissue defects caused by trauma, such as bone defects in the distal leg, nonunion, large necrotic space as well as traumatic surface of the foot and ankle. PMID- 19546681 TI - Should we hesitate to use subcutaneous tunneling for fear of damaging the sural flap pedicle? AB - BACKGROUND: The distally based sural nerve flap is a preferable option for covering defects of the lower third of the leg. However, many authors noted that in particular pressure exerted by tunneling of the flap pedicle could be harmful. Thus, they either inset the pedicle or exteriorize the pedicle to decrease complications. METHODS: We used distally based superficial sural artery island flaps for the reconstruction of defects of the lower leg, the ankle, and malleolus in 12 patients between 2004 and 2008 without tunelization. The defects were covered with no major complications. RESULTS: In 3 flaps, there was a small amount of distal marginal necrosis, which was excised and closed spontaneously or skin grafted. CONCLUSION: We concluded that unless there are certain risk factors such as arterial hypertension, diabetes, peripheral vascular disease, and vasculitis, avoiding a subcutaneous tunneling is not necessary. PMID- 19546682 TI - Effect of camptothecin on collagen synthesis in fibroblasts from patients with keloid. AB - Keloids are distinguished by substantial deposition of collagen in the dermis, resulting in an imbalanced production and aggregation of extra cellular matrix. This study was undertaken to evaluate the effects of the topoisomerase I inhibitor camptothecin (CPT) on collagen synthesis in the activated dermal fibroblasts from healthy donors and patients with keloid. The fibroblasts were cultured in the presence or absence of CPT. Cellular toxicity assay was determined by MTT analysis. The expression of type I collagen and type III collagen was studied both at the transcriptional and post-transcriptional levels, using conventional quantitative real-time reverse transcription PCR and Western blotting. Results showed that there was predominantly a clear and dose-dependent decrease in the synthesis of collagen 1, not collagen 3, in keloid fibroblasts without significantly cellular toxicity. The CPT had an activity on the regulation of the ratio of type I/III collagen in the metabolism of keloid fibroblasts by inhibiting the secretion of type I collagen. The data suggest that the inhibitory effect of CPT, a topoisomerase I inhibitor, on collagen synthesis may be an effective treatment for limiting fibrosis in keloid patients. PMID- 19546683 TI - Effect of cyclooxygenase-2 on ischemic preconditioning of skin flaps. AB - Ischemic preconditioning is a useful tool to fight against reperfusion injury. This phenomenon is very complex and the underlying mechanism has various branches. Every study on ischemic preconditioning helps us to better understand this process. We aimed to investigate the effectiveness of cyclooxygenase-2 (COX 2) on ischemic preconditioning of skin flaps in the rat. A 6 x 3 cm-sized left epigastric artery flap was used and the pedicle was isolated to perform the ischemic preconditioning via microvascular clamp application. The preconditioning protocol was 2 cycles of 15 minutes ischemia and 15 minutes reperfusion periods. Sixty female Wistar rats weighing between 210 and 260 g were used for the experiment. Animals were allocated randomly into 6 groups, each group containing 10 animals. Group 1: Only 6 hours of ischemia was done after the flap elevation, neither ischemic preconditioning nor COX-2 inhibitor was used; Group 2: 6 hours of global ischemia was induced just after the ischemic preconditioning; Group 3: In addition to the same procedures in group 2, 2 doses of COX-2 inhibitor were given before and after the final ischemic insult; Group 4: 6 hours of ischemia was applied to the flap 24 hours after its elevation, no preconditioning or any other interventions were done; Group 5: The same ischemic protocol was used after the flap elevation but the 6 hours of ischemia was performed 24 hours after the preconditioning; Group 6: The same procedures of group 5 were done and in addition, 2 doses of COX-2 inhibitor was given, starting 24 hours after the ischemic preconditioning. All flaps were followed for 1 week then necrotic flap portions were measured and represented as a percentage to the whole flap area. Statistical analyses revealed meaningful differences between groups 2 and 3 (P < 0.05), 2 and 1 (P < 0.05), 5 and 6 (P < 0.05), 5 and 4 (P < 0.05). However, there was no statistical difference between groups 3 and 1 (P > 0.05), 6 and 4 (P > 0.05). As a conclusion, ischemic preconditioning has both early and late protective effects on ischemia-reperfusion injury in the skin flap model. By antagonizing COX-2 receptors the beneficial effects of IP were reversed. This result indicated that COX-2 has a specific role in the mechanism of both early and late effects of ischemic preconditioning in skin flaps. PMID- 19546684 TI - Plastic surgery within the Veterans Affairs Medical System: proposed modified indications for operative procedures. AB - Many plastic surgery procedures span the divide between aesthetic ("cosmetic") and reconstructive surgery. However, definitions and guidelines may be inconsistent, which may decrease patients' access to legitimate procedures. The article aims to assist Veterans' Health Administration-affiliated plastic surgeons in continuing to provide optimal care to the Nation's Veterans and family members, and should be regarded as an open discussion. PMID- 19546685 TI - Dangers of cornstarch powder on medical gloves: seeking a solution. AB - This article reviews information on the hazards of cornstarch powder on medical gloves. Dusting powders were first applied to latex gloves to facilitate donning. After 1980, manufacturers devised innovative techniques without dusting powder. It has been well documented that these powders on gloves present a health hazard to patients and health care workers by 5 different mechanisms. First, the glove cornstarch has documented detrimental effects on wound closure techniques. Second, this powder potentiates wound infection. Third, cornstarch induces peritoneal adhesion formation and granulomatous peritonitis. Finally, these powders serve as carriers as latex allergen and they precipitate a life threatening allergic reaction in sensitized patients. These well-documented hazards of glove powder have caused the United Kingdom and Germany to ban cornstarch powder on medical gloves over 10 years ago. PMID- 19546687 TI - When and how did Giulio Cesare Arantius (1530-1589) learn to perform nasal reconstructions? PMID- 19546688 TI - Hingson's peace gun. PMID- 19546689 TI - The physician's pulse watch. PMID- 19546691 TI - Impact of gene copy number variation on anesthesia in Drosophila melanogaster. AB - BACKGROUND: Chromosomal deletions and duplications, which result in halving or doubling of copy number in a block of genes, are an important source of variation between individuals. Phenotypic effects of copy number variation are commonly observed, but effects on sensitivity to volatile anesthetics have not been assessed in any organism. METHODS: The potency with which halothane depresses the righting reflex of fruit flies was measured in congenic Drosophila strains, each of which was heterozygous for a deletion of average size 400 kb. Over 200 strains were examined, thereby scanning approximately half of the fly genome. RESULTS: Although the vast majority of deletion heterozygotes were indistinguishable from the control, eight had significantly altered sensitivity to halothane. Genetic tests supported the hypothesis that the change in anesthetic sensitivity was the result of reduction in copy number and not adventitious mutations in the strains. Among the eight outliers, the difference in halothane potency ranged from a 25% increase to a 15% decrease. Changes of similar magnitude but distinctive patterns were found when these lines were tested with enflurane, isoflurane, and sevoflurane. CONCLUSIONS: Variation in gene copy number has a significant impact on anesthetic sensitivity in Drosophila melanogaster. The level of transcription of a few genes must thus be limiting for a normal response to volatiles. Coupling between gene copy and gene expression is universal, and the components of the fly's nervous system are highly conserved; therefore, this work provides a rationale for investigating the clinical impact of copy number variation. PMID- 19546692 TI - Preoperative melatonin and its effects on induction and emergence in children undergoing anesthesia and surgery. AB - BACKGROUND: Studies conducted in adults undergoing surgery reported a beneficial effect of oral melatonin administered before surgery. There is a paucity of such data in children undergoing anesthesia and surgery. METHODS: Children undergoing surgery were randomly assigned to receive preoperatively oral midazolam 0.5 mg/kg or oral melatonin 0.05 mg/kg, 0.2 mg/kg, or 0.4 mg/kg. The primary outcome of the study was preoperative anxiety (Yale Preoperative Anxiety Scale). The secondary outcomes were the children's compliance with induction (Induction Compliance Checklist), emergence behavior (Keegan scale), and parental anxiety (State-Trait Anxiety Inventory). RESULTS: Repeated measures ANOVA showed that children who received melatonin at any of the three doses were more anxious compared with children who received midazolam (P < 0.001). Parental anxiety did not differ on the basis of the experimental condition (P = ns). The melatonin groups showed a dose-response effect on emergence behavior. Children who received melatonin developed less emergence delirium compared with those who received midazolam (P < 0.05), and the effect was dose related; the incidence after 0.05 mg/kg melatonin was 25.0%, incidence after 0.2 mg/kg melatonin was 8.3%, and incidence after 0.4 mg/kg melatonin was 5.4%. CONCLUSIONS: Midazolam is more effective than melatonin in reducing children's anxiety at induction of anesthesia. Melatonin showed a direct dose-dependent effect on emergence delirium. PMID- 19546693 TI - Kirschbaum's oxygen content controller. PMID- 19546695 TI - Retrospective but not rigorous. PMID- 19546697 TI - Surgical demand time variance: a possible explanation for lunar effects and some other mysterious cycles. PMID- 19546699 TI - Perioperative statin therapy may be implicated in a wide array of drug-drug interactions. PMID- 19546700 TI - Oscillations in the plethysmographic waveform amplitude: phenomenon hides behind artifacts. PMID- 19546703 TI - Chloral hydrate is not acceptable for anesthesia or euthanasia of small animals. PMID- 19546704 TI - Low-dose spinal anesthesia with low-dose phenylephrine infusions for cesarean delivery: better but not necessarily best. PMID- 19546705 TI - Maternal hemodynamic monitoring and the Vigileo monitor. PMID- 19546707 TI - Low-dose versus a higher-dose bupivacaine spinal anesthesia for cesarean delivery. PMID- 19546709 TI - Muscle relaxants and airway management. PMID- 19546710 TI - Is faculty presence during emergent tracheal intubations justified? PMID- 19546717 TI - Diabulimia: what it is and how to recognize it in critical care. AB - Critical care nurses must be able to recognize the signs of symptoms of diabulimia-a potentially life-threatening disorder. Skipping insulin is used as a means of weight control in some persons with diabetes, particularly in young women. This article focuses on the assessment, pathophysiology, critical care nursing interventions, and psychosocial initiatives of interest to critical care nurses in the care of patients with diabulimia. PMID- 19546719 TI - Cardiopulmonary resuscitation training for family members. AB - Out-of-hospital sudden cardiac arrest survival rate is significantly higher when the patient receives cardiopulmonary resuscitation (CPR) before the arrival of emergency providers although published data suggest a significantly lower prevalence of CPR training among cardiac patients' family members and bystanders in the United States. This article presents information including attitudes of patients, family members, and the general population toward CPR; the barriers for CPR training among the public; and the most effective method of CPR training for bystanders, so that CPR training for family members can be promoted to improve out-of-hospital sudden cardiac arrest survival rate. PMID- 19546721 TI - The effects of depression in heart failure. AB - Depression is commonly seen in patients with heart failure. Studies have shown an increase in hospitalizations and healthcare costs. Patients with depression are twice as likely to die prematurely. However, the diagnosis and treatment of depression in heart failure are difficult because of the similarity of symptoms. Early detection of depression in these patients will increase their quality of life and overall survival. PMID- 19546722 TI - Innovative solutions: family-centered care visitation guidelines. AB - This brief article describes one unit's efforts in establishing family-centered care visitation in an effort to better meet patient and family needs. PMID- 19546724 TI - Innovative solutions: optimal patient outcomes as a result of multidisciplinary rounds. AB - Multidisciplinary rounds have been shown to improve patient outcomes. The use of a standardized tool in one institution led to a pharmacy cost savings of more than 50,000 dollars and a decrease of 1.5 days for patients receiving mechanical ventilation. PMID- 19546727 TI - Psychological impact of implantable cardioverter defibrillator on their recipients. AB - This 12-month longitudinal prospective study tracked the depression and anxiety levels of implantable cardioverter defibrillator recipients after implantation of the device. Depression and anxiety levels dropped significantly over time. These findings suggest that interventions should be implemented early because the needs are greatest in the first few weeks after device implantation. PMID- 19546729 TI - Intensive care unit nurse characteristics impacting judgments about secondary brain injury. AB - Although intensive care unit nurses have an integral role in preventing secondary brain injury when caring for critically ill traumatic brain injury patients, nursing practice varies and little research has examined if individual nurse characteristics affect judgments about secondary brain injury. The purpose of this study was to investigate how intensive care unit nurse characteristics influence judgments when managing secondary brain injury. A factorial survey research design was used to administer surveys to intensive care nurses from 2 level I trauma centers. Nursing shift and number of years in the intensive care unit were significant predictors of judgments. Understanding the role of nurse characteristics is important in future educational and research efforts aimed at evidence-based management of secondary brain injury. PMID- 19546730 TI - Perceptions of recovery, physical health, personal meaning, role function, and social support after first-time coronary artery bypass graft surgery. AB - Coronary artery bypass graft surgery is one of the most frequently performed major surgeries in the United States. Because of fast-track protocols and decreased lengths of stay, critical care nurses have even more demanding challenges in the care of these patients. Critical care nurses are well aware that these patients still have health needs that extend beyond discharge from the hospital, but these needs are now addressed by the patient's caregiver. This descriptive, comparative study was conducted to examine perceptions of recovery by these patients and the relationships among physical health, personal meaning, role function, and social support. The results of the study are presented, as well as the implications for critical care nurses. PMID- 19546733 TI - A guest editorial: 7 tips for critical care nurses to take back nursing. PMID- 19546735 TI - From the editor. Innovation in transforming organizations. PMID- 19546736 TI - Guest editorial. Innovations in transforming organizations. PMID- 19546737 TI - Letter on "Creating a healing environment: an innovative educational approach for adopting Jean Watson's theory of human caring". PMID- 19546738 TI - Creating a context for excellence and innovation: comparing chief nurse executive leadership practices in magnet and non-magnet hospitals. AB - Chief nurse executives create a context for leadership, innovation, and practice in hospitals. It is valuable to get a sense of nurse executives' perceptions regarding their leadership practices and how they value them. Furthermore, it is of interest to see if there is significant differentiation in these perceptions between chief nurse executives in Magnet hospitals and those in non-Magnet hospitals. This article discusses a study of the leadership practices of these 2 groups of nurse executive's leadership practices and reports the results. Concluding is a brief discussion regarding impact and importance of the nurse executive related to excellence and innovation. PMID- 19546739 TI - Teaching innovation. AB - Innovation in healthcare is essential to solve the "wicked problems" currently facing healthcare. This article focuses on nature of innovation and how it operates, how innovators think and view problems, how the theory and practice of innovation can be taught in novel ways, and how organizational cultures foster or suppress innovation. Examples of teaching strategies and nurse-driven innovation illustrate the theory and practice of innovation. PMID- 19546740 TI - Therapeutic benefits of expressive writing in an electronic format. AB - Expressive writing is recognized as having therapeutic benefits including fewer stress-related visits to the doctor, improved immune system functioning, reduced blood pressure, improved functional status, fewer days in the hospital, feelings of greater psychological well-being, reduced depressive symptoms, fewer posttraumatic intrusion and avoidance symptoms, improved memory and sleep, and faster healing after surgery. Combine this with current popularity of social networking, and there is a formula for improved patient outcomes. PMID- 19546741 TI - Diversity within nursing: effects on nurse-nurse interaction, job satisfaction, and turnover. AB - Little research has focused on understanding interpersonal dynamics within a diverse nursing staff. This study investigated the impact of diversity on the interaction level among staff nurses, job satisfaction, nursing turnover, and the multicultural sensitivity of a diverse nursing staff in metropolitan Washington, DC. Data were collected from 194 registered nurses with 2 standardized instruments--the Workforce Diversity Questionnaire-II by Larkey and the Multicultural Sensitivity Scale Questionnaire by Jibaja-Rusth and others. One-way analyses of variance and correlation coefficient were used for data analysis. The findings showed that nurses who were satisfied with their current job were more likely to value differences and build trusting relationships. Nurses with higher educational levels appeared to be more open and involved with other cultural groups and were more likely to build more trusting relationships with other cultural groups. Multicultural sensitivity was related to cultural group inclusion/exclusion, valuing differences, and adaptation; however, multicultural sensitivity and trust were not related. Delivery of cost-effective, quality nursing care in the 21st century demands that the positive potential of cultural diversity in the nursing workforce be more fully understood through research such as this. More research on diversity is needed during these challenging times for the nursing profession. PMID- 19546742 TI - Innovation in healthcare: a concept analysis. AB - Innovation is a frequently used buzzword in healthcare. This article will clarify innovation as a process requiring leadership, among other factors, in order to occur. The concept of innovation will be defined, as well as the precedents and consequences. This exploration will serve as the definition of healthcare innovation and provide a clearer definition for future literature and research in healthcare, especially related to leadership and change. It is the purpose for this article to allow the reader to think about innovation in a critical manner and begin to add substantive meaning related to it. PMID- 19546743 TI - Engaging the nursing workforce: an evidence-based tool kit. AB - Hand hygiene is critical to the safety of patients in hospitals today. OBJECTIVE: Engaging staff, physicians, patients, and visitors in hand hygiene is important to create a safe environment. The author discusses the development and implementation of an evidence-based practice guideline for improving hand hygiene and application of that guideline in an academic medical center setting. METHOD: Establishment of a multidisciplinary team committed to improving organizational performance, development of a practice guideline, organizational assessment, and the implementation of a multimodal intervention plan are all keys to success. CONCLUSION: Organizational culture can be changed utilizing an evidence-based, organization-specific approach to hand hygiene. PMID- 19546745 TI - Implementing peer review: guidelines for managers and staff. AB - The performance appraisal process is a key component of professional nursing practice. The peer review process is referred to frequently in Magnet Nursing Standards as a key element in professional nursing. The peer review process allows professionals from common practice areas to systematically assess, monitor, make judgments, and provide feedback to peers by comparing actual practice to established standards. Peer review can engage a multigenerational workforce and lead to more satisfied, engaged employees. As a component of the annual performance appraisal, peer review can create positive relationships, foster a better work environment, and allow peers to increase individual and group accountability. Peer review has many common elements that can be individualized to fit any type of unit or work culture. This article describes how to design, implement, and evaluate a unit-based peer review program. The content and steps outlined are intended to support nurse managers in implementing unit-specific peer review programs by focusing on existing expertise and best practices. The implementation steps are divided into 5 phases describing the introduction of concepts and getting nurses engaged, implementation guidelines, piloting the process, staff education, and ongoing evaluation. Staff involvement is the key to a successful unit-based peer review process. PMID- 19546744 TI - A successful online mentoring program for nurses. AB - AIM: This article describes the successful implementation of An Online Mentoring Program for Nurses at a Magnet-designated acute care medical center, Morristown Memorial Hospital (MMH/Atlantic Health). BACKGROUND: A comprehensive approach to incorporating mentor-protegee teams into professional nurse role development has been demonstrated to (1) improve nurse employee satisfaction, retention, and recruitment outcomes; (2) change the ways nurses and others perceive nurses; (3) augment support by managers and coworkers; and (4) improve patient care outcomes. EVALUATION: Nurses are partnered in mentor-protegee relationships and continually engage one another by evaluating the protegee's unique contributions and identifying specific strategic actions to move the protegee toward accomplishing their professional objectives. KEY ISSUES: Building an online mentor-protegee collaboration: (1) maximizing potential, (2) identifying the protegee's unique contributions, and (3) strategic planning. CONCLUSIONS: The online mentoring process is a success and has delivered measurable results that have benefited the nurse participants and contributed to our institution's culture of nursing engagement. IMPLICATIONS FOR NURSING MANAGEMENT: The online mentoring process has potential to benefit nurses and their organizations by (1) providing real-time communication, (2) facilitating strategic thinking, (3) monitoring progress, (4) "going green," and (5) improving organizational knowledge. PMID- 19546746 TI - Climate for innovation: problems in patient flow. AB - Problems in patient flow persist in practice and seem to defy traditional approaches to resolution. Using concepts from complex adapted systems and diffusion of innovation principles, problems in patient flow are analyzed and possible routes for innovation are identified. Emphasis is on creating a climate receptive to change as the norm, one that values relationships and communication. PMID- 19546747 TI - Innovations in regulation. PMID- 19546748 TI - Innovations in transforming organizations. AB - Transformations take as many forms as the organizations they change. However, the success of healthcare-based transformations requires significant cultural shifts and operational changes, many of which mirror the tenets of classic transformer W. Edward Deming. Attaining Magnet hospital status and the integration of the e health record into nursing school curricula are 2 examples of how healthcare leaders have championed a series of changes that produced sweeping transformations. Success is likely to elude healthcare leaders who mount massive transformations, while those who focus on making seemingly small, interconnected changes are more likely to see the results they envision over time. PMID- 19546750 TI - New developments in the surgical management of early endometrial cancer. PMID- 19546751 TI - "Transitions". PMID- 19546752 TI - Evaluation of two management strategies for preoperative grade 1 endometrial cancer. AB - OBJECTIVE: To compare the practices, adjuvant treatment, and outcomes of patients with preoperatively assessed grade 1 endometrioid endometrial cancer between two academic gynecologic oncology centers that use different treatment strategies. METHODS: A retrospective analysis was performed at Duke University Medical Center (Duke) and the Toronto Sunnybrook Regional Cancer Center (Sunnybrook) between 1991 and 2007. Patients at Duke generally underwent surgical staging unless intraoperative assessment identified a negligible risk of nodal disease. Patients at Sunnybrook generally did not undergo surgical staging. RESULTS: A total of 494 patients (272 from Duke and 222 from Sunnybrook were identified with preoperative, central-review-confirming, grade 1, endometrioid, endometrial cancer. Groups were similar in grade, final histology, type of hysterectomy, and length of hospital stay. Patients from Sunnybrook were older (aged 62 years compared with 59 years, P=.001) and were more likely to have capillary lymphatic space involvement (18.2% compared with 8.3%, P=.003) and cervical involvement (12.2% compared with 3.7%, P<.001). Approximately 2% of cases were upgraded to high grade on final specimen. Lymphadenectomy was performed on 49.4% of patients at Duke compared with 11.7% of patients at Sunnybrook. Overall 3-year survival was 96% at Duke and 96% at Sunnybrook (P=.217). Three-year recurrence-free survival was 96% at Duke and 95% at Sunnybrook (P=.327). CONCLUSION: Despite differences in practice and slight differences in patient populations, the recurrence-free and overall survival of women with preoperative centrally reviewed grade 1 endometrial cancer is excellent and without statistically significant difference between the two centers. LEVEL OF EVIDENCE: III PMID- 19546753 TI - Comprehensive surgical staging for endometrial cancer in obese patients: comparing robotics and laparotomy. AB - OBJECTIVE: To compare adequacy and outcomes of surgical staging for endometrial cancer in obese women by robotics or laparotomy. METHODS: Clinical stage I or occult stage II endometrial cancer patients with body mass indexes (BMIs) of at least 30 (BMI is calculated as weight (kg)/[height (m)]2) were identified undergoing robotic staging and matched 1:2 with laparotomy patients. Patient characteristics, operative times, complications, and pathologic factors were collected. An adequate lymphadenectomy was defined arbitrarily as at least 10 total nodes removed, and adequate pelvic and paraaortic lymphadenectomy was defined as at least six and at least four nodes removed, respectively. RESULTS: A total of 109 patients underwent surgery with the intent of robotic staging and were matched to 191 laparotomy patients. The mean BMI was 40 for each group. The robotic conversion rate was 15.6% (95% confidence interval [CI] 9.5-24.2%). Ninety-two completed robotic patients were compared with 162 matched laparotomy patients. The two groups were comparable regarding total lymph node count (25 +/- compared with 24 +/- 12, P =.45) and the percentage of patients undergoing adequate lymphadenectomy (85% compared with 91%, P=.16) and adequate pelvic (90% compared with 95%, P=.16) and aortic lymphadenectomy (76% compared with 79%, P=.70) for robotic and laparotomy patients, respectively, but there was limited power to detect this difference. The blood transfusion rate (2% compared with 9%, odds ratio [OR] 0.22, 95% CI 0.05-0.97, P=.046), the number of nights in the hospital (1 compared with 3, P<.001), complications (11% compared with 27%, OR 0.29, 95% CI 0.13-0.65 P=.003), and wound problems (2% compared with 17%, OR 0.10, 95% CI 0.02-0.43, P=.002) were reduced for robotic surgery. CONCLUSION: In obese women with endometrial cancer, robotic comprehensive surgical staging is feasible. Importantly, obesity may not compromise the ability to adequately stage patients robotically. LEVEL OF EVIDENCE: II PMID- 19546754 TI - Endometrial cancer risk among younger, overweight women. AB - OBJECTIVE: To examine the risk for endometrial cancer among overweight women using the World Health Organization's clinical definitions of obesity based on body mass index (BMI). METHODS: Conducted in the early 1980s, the Cancer and Steroid Hormone study was a multicenter, population-based, case-control study of breast, ovarian, and endometrial cancers among women aged 20-54 years. Participants for the case group (n=421) were identified through cancer registries and had histologically confirmed endometrial cancer. Participants for the control group (n=3,159) were chosen by random-digit dialing methods in the same regions as those in the case group. Those in the case and control groups responded to the same questions during in-person interviews. Unconditional logistic regression was used to estimate adjusted odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: The relationship between endometrial cancer and BMI (calculated as weight [kg]/[height (m)]) was modified by age at last menstrual period (LMP). Of women who were younger than 45 years at LMP, those with BMIs of at least 35.0 had a greater risk of endometrial cancer (56%, 30/54) than did those with normal BMIs (4%, 59/1,492, adjusted OR 21.7, 95% CI 11.3-41.7). Of women age 45 or older at LMP, those with BMIs of at least 35.0 also had a greater risk (40%, 24/60) than did those with normal BMIs (14%, 168/1,235, adjusted OR 3.7, 95% CI 2.0-6.6). Women younger than 45 years at LMP and those with BMIs of at least 25.0 at 18 years and as adults (25%, 31/123) had an approximately sixfold increased risk (adjusted OR 5.8, 95% CI 3.4-9.8) compared with those with normal BMIs at 18 and as adults (4%, 58/1,460). CONCLUSION: Very obese women aged 20-54 years have an elevated endometrial cancer risk, which appears heightened by early menopause. LEVEL OF EVIDENCE: II. PMID- 19546755 TI - Outcomes of expectantly managed preterm premature rupture of membranes occurring before 24 weeks of gestation. AB - OBJECTIVE: To assess contemporary outcomes in expectantly managed preterm premature rupture of membranes (PROM) before 24 weeks of gestation. METHODS: We analyzed all patients with singleton pregnancies and preterm PROM before 24 weeks of gestation from 2001 to 2007. Patients immediately electing delivery, delivering within 12 hours of preterm PROM, carrying anomalous fetus(es), or multiple gestations were excluded. Neonatal survival without major morbidities was the primary outcome. Data were analyzed with multivariable logistic regression and Cox regression models. Week-specific probability estimates for neonatal morbidity and mortality were calculated based on gestational age at the time of preterm PROM. RESULTS: One hundred fifty-nine women fulfilled study criteria. Median gestational age at preterm PROM for all patients was 21.4 (range 14.0-23.9) weeks of gestation. Median delivery gestational age was 24.7 (range 15.4-34.1) weeks. Forty-seven patients experienced either an intrauterine fetal demise, elected delivery after initial expectant management, or delivered before planned resuscitation. Of 112 newborns admitted to neonatal intensive care, 89 (56.0% of all neonates) survived; 43 (48.3% of survivors, 27.0% of all neonates) had no major neonatal morbidities. Morbidity probabilities decreased with increasing gestational age at the time of preterm PROM. Delivery gestational age was predictive of both neonatal morbidity and mortality. CONCLUSION: More than one half of women who achieved at least 12 hours of latency and elected expectant management had a surviving infant; nearly 50% of survivors had no major neonatal morbidity. These contemporary outcomes are valuable in counseling women with early preterm PROM. LEVEL OF EVIDENCE: III. PMID- 19546757 TI - Effect of 17alpha-hydroxyprogesterone caproate on glucose intolerance in pregnancy. AB - OBJECTIVE: To estimate whether 17alpha-hydroxyprogesterone caproate treatment in pregnancy increases the frequency of abnormal glucose screening and gestational diabetes mellitus (GDM). METHODS: This is a retrospective cohort study of women treated with weekly 17alpha-hydroxyprogesterone caproate. Women with pregestational diabetes and multiple gestations were excluded. 17alpha hydroxyprogesterone caproate-exposed women were randomly matched with three unexposed controls by maternal age and prepregnancy body mass index (BMI). The main outcomes were an abnormal 1-hour 50-g glucose screen (at least 135 mg/dL) and GDM (a 1-hour 50-g glucose screen of at least 200 mg/dL or two or more abnormal values on a 3-hour 100-g oral glucose tolerance test). RESULTS: A total of 110 17alpha-hydroxyprogesterone caproate-exposed women were matched with 330 controls. Maternal race between exposed women and controls was similar (46% compared with 39% African American, 17% compared with 18% Hispanic, 36% compared with 40% white, P=.57). Abnormal 1-hour glucose screens were more frequent in the 17alpha-hydroxyprogesterone group (23.6% compared with 11.2%, P<.001), as was the diagnosis of GDM (10.9% compared with 3.6%, P=.003). 17alpha-hydroxyprogesterone caproate remained independently associated with the diagnosis of GDM (odds ratio 3.3, 95% confidence interval 1.3-8.1) in a conditional multiple logistic regression analysis controlling for maternal race, age, BMI, and parity. CONCLUSION: Women receiving weekly intramuscular 17alpha-hydroxyprogesterone caproate have more frequent abnormal glucose testing and gestational diabetes compared with unexposed controls. These results are consistent with published data regarding the effect of progesterone on insulin resistance. LEVEL OF EVIDENCE: II. PMID- 19546756 TI - Bone metabolism in fetuses of pregnant women exposed to single and multiple courses of corticosteroids. AB - OBJECTIVE: To estimate the effect of single and recurrent doses of antenatal corticosteroids on fetal bone metabolism. METHODS: This was a secondary analysis of a cohort of pregnant women from a previously reported randomized, placebo controlled, multicenter trial of women at risk for preterm delivery who received weekly courses of betamethasone (active) or placebo after an initial course of corticosteroids. Umbilical cord serum levels of carboxy-terminal carboxy-terminal propeptide of type I procollagen and cross-linked carboxy-terminal telopeptide of type I procollagen were measured to assess the rate of fetal bone formation and resorption, respectively. Analysis was stratified according to number of repeat antenatal study courses of betamethasone or placebo (one to three compared with at least four courses, not including the initial course). RESULTS: Of the 251 umbilical cord serum samples, the median serum carboxy-terminal telopeptide of type I procollagen levels, but not carboxy-terminal propeptide of type I procollagen levels, was significantly lower with repeat betamethasone exposure (55.0 compared with 57.9 micrograms/L, P=.01). In the fetuses exposed to at least four repeat study courses, there was a significant decrease in median carboxy terminal telopeptide of type-I procollagen levels between repeat betamethasone exposure and placebo (53.4 compared with 58.6 micrograms/L, respectively, P=.04), but there was no difference between groups in the fetuses exposed to 1-3 repeat study courses (57.4 compared with 56.7 micrograms/L, respectively, P=.29). CONCLUSION: Levels of umbilical cord serum markers of bone resorption but not formation are reduced in fetuses exposed to repeat courses of antenatal betamethasone. Up to four courses of antenatal betamethasone do not seem to affect fetal bone metabolism. LEVEL OF EVIDENCE: II. PMID- 19546758 TI - Triple-marker prenatal screening program for chromosomal defects. AB - OBJECTIVE: To examine screening performance of California's triple-marker screening program, using data from a statewide registry for chromosomal defects. METHODS: This study included 752,686 women who received a screening risk and had an expected date of delivery between July 2005 and the end of June 2007. Follow up diagnostic services for screen-positive women were performed at state-approved centers. Data on diagnostic outcomes from these visits were entered into the California Chromosomal Defect Registry (CCDR). Other CCDR sources include mandatory reporting by all cytogenetic laboratories and hospitals and outcome data forms submitted by prenatal care providers. RESULTS: The observed detection rate for Down syndrome (N=1,217) was 77.4%. It varied significantly by gestational dating method and maternal age. The rates for women aged younger than 35 years and 35 years and older were 62.4% and 94.0%, respectively. The detection rates were 81.3% for ultrasound-dated pregnancies and 67.5% for last menstrual period-dated pregnancies. For Turner syndrome, trisomy 18, triploidy, and trisomy 13, the detection rates were 79.4%, 82.5%, 98.1%, and 36.0%, respectively. The positive rate for Down syndrome was 5.4%. Of women with a Down syndrome fetus who were screen positive, only 49.5% opted for amniocentesis. Of women who obtained results from amniocentesis indicating a Down syndrome fetus, 61.4% had an elective termination, 26.2% had a live birth, 4.5% had a death or miscarriage, and 7.9% had an unknown outcome. CONCLUSION: The observed performance of this large triple-marker screening program exceeds generally predicted detection rates for Down syndrome. This study methodology will be used to measure the performance of subsequent screening enhancements. LEVEL OF EVIDENCE: III. PMID- 19546759 TI - Anterior repair with or without collagen matrix reinforcement: a randomized controlled trial. AB - OBJECTIVE: To compare outcomes of anterior colporrhaphy alone to that reinforced with bovine pericardium graft. METHODS: Women with anterior vaginal wall prolapse were enrolled in a randomized fashion in this grafted compared with nongrafted repair study. Outcome measures included pelvic organ prolapse quantification data, quality-of-life assessment, healing abnormalities, and complications. RESULTS: Ninety-four patients were enrolled. Seventy-two (77%) provided 1-year data, and 59 (63%) supplied 2-year data. Demographics and stage of prolapse were similar between groups at baseline. Postoperative complications consisted basically of low urinary tract infection and were low in both groups (10 in bovine pericardium graft and 16 in anterior colporrhaphy alone). One year after surgery, successful anterior vaginal wall support was obtained in 85.7% of the bovine pericardium graft group and 78.4% of anterior colporrhaphy-alone group (P=.544). For the cohort that comprised 2-year analyses, the success rate was 76.5% for the bovine pericardium graft group and 63% for anterior colporrhaphy alone group (P=.509). Postoperative Urogenital Distress Inventory-6 and Pelvic Organ Prolapse-Urinary Incontinence Sexual Function Questionnaire-12 scores were uniformly improved over baseline in both groups. CONCLUSION: The use of bovine pericardium graft for anterior vaginal prolapse does not have higher complication rates or healing difficulties. At 1- and 2-year follow-up, anterior colporrhaphy with bovine pericardium reinforcement did not show a statistically significant improvement over colporrhaphy alone. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00860912 LEVEL OF EVIDENCE: I. PMID- 19546760 TI - Appearance of the levator ani muscle subdivisions in endovaginal three dimensional ultrasonography. AB - OBJECTIVE: To formulate an evaluation system and authenticate the levator ani subdivisions visible on endovaginal three-dimensional ultrasonography. METHODS: A three-dimensional endovaginal ultrasound examination was performed on five fresh frozen female pelves. The location of the pubovaginalis, puboperinealis, puboanalis, puborectalis, and iliococcygeus muscles was identified. Three dimensional endovaginal ultrasound scans of 22 nulliparous women with normal pelvic floors were obtained, and a three-level evaluation system was constructed. Level 1 contained the muscles as they insert into the perineal body. Level 2 evaluated the insertions into the retropubic area and typically contained the pubovaginalis, puboperinealis, puboanalis, and puborectalis. Level 3 contained muscles that were cephalad to the pubic bone. Two blinded reviewers assessed levator ani subdivision visualization at each level and individual muscle visualization by the origin-insertion points. RESULTS: There was 98%, 96%, and 92% agreement for levels 1, 2, and 3 muscles, with 95% confidence intervals (CIs) of 0.92-1, 0.95-0.99, and 0.88-0.95, respectively. Kappa values (95% CI) for agreement were calculated for individual muscles as follows: superficial transverse perinei and puborectalis were seen by both raters 100%, puboperinealis, pubovaginalis, and puboanalis 0.645 (0.1-1), and iliococcygeus 0.9 (0.6-1). CONCLUSION: The location of the pubovaginalis, puboperinealis, puboanalis, puborectalis, and iliococcygeus as seen by three-dimensional endovaginal ultrasonography was confirmed through anatomic dissection of fresh frozen pelves. Subdivisions of the levator ani muscle were visualized reliably with three-dimensional ultrasonography using a systematic approach. LEVEL OF EVIDENCE: III. PMID- 19546761 TI - Route of hysterectomy influence and teaching hospital status. AB - OBJECTIVE: The purpose of this study was to evaluate whether teaching hospitals are more likely to perform abdominal (laparotomy) compared with nonabdominal (vaginal, laparoscopic) hysterectomies for benign indications. METHODS: We conducted a retrospective, observational study comparing abdominal and nonabdominal inpatient hysterectomies performed in Illinois between 2000 and 2005 using the COMPdata database. Obvious cancer, prolapse, or indicated-abdominal surgeries (infection and pregnancy-related cases) were excluded. The final analysis included 94,599 cases. Diagnoses and patient demographics were analyzed from the database and hospitals' teaching status, as determined by telephone interviews. The relationship between route of hysterectomy and teaching hospital status was modeled using multivariable logistic regression with a P value cutoff of less than 0.05. RESULTS: Eight-two percent of hysterectomies performed at teaching hospitals were performed abdominally compared with 77% at nonteaching hospitals. After adjusting for age and diagnoses, teaching hospitals were less likely to perform hysterectomy by abdominal approach (odds ratio 0.69, 95% confidence interval 0.49-0.97, P<.035). Clinical variables associated positively with vaginal hysterectomies included primary diagnoses of menstrual disorders, other female genital disorders, and menopausal disorders. Complication rate did not differ by teaching status. Laparoscopic hysterectomy, even after adjustment for confounders, was associated with fewer complications compared with both abdominal and vaginal routes. CONCLUSION: The route of hysterectomy is only minimally influenced by teaching hospital status. These findings are important for clinician-educators responsible for teaching the nation's next generation of gynecologic surgeons. Strategies to overcome presumed physician-level factors are needed to optimize patient outcomes through appropriate use of nonlaparotomy surgery. LEVEL OF EVIDENCE: III. PMID- 19546762 TI - Adolescent sexual behavior during periods of increase and decrease in the abortion rate. AB - OBJECTIVE: To study changes in adolescent sexual behavior in periods of increase (1994-2000) and decrease (2001-2007) in the abortion rate. METHODS: School surveys with self-administered questionnaires were carried out annually among eighth graders (mean age 14.8 years) and ninth graders (mean age 15.8 years) (N=286,665) in 1996/1997 and 2006/2007. Schools participated biennially. The proportions of respondents reporting noncoital (kissing, light petting, heavy petting) and coital (ever had sexual intercourse, intercourse at least 10 times, at least three partners) sexual experience and nonuse of contraception were studied. RESULTS: Among adolescents, both coital and noncoital sexual experiences and the proportion of those not using contraception increased between 1996-1997 and 2000-2001 (P for trend <.01, all) and decreased from 2000-2001 onward (P<.001, all), except the proportion of at least 10 coital events, which did not decrease. Among sexually experienced adolescents, a similar increase in coital experiences (intercourse at least 10 times from 38.2% to 41.5%, at least three partners from 27.8% to 30.7%, P<.001) and in not using contraception (from 17.2% to 19.1%, P=.002) was seen before 2000-2001, but after that the only significant change was a further increase in the proportion of those reporting intercourse at least 10 times (from 41.5% to 47.8%, P<.001). CONCLUSION: The proportion of adolescents reporting noncoital sexual experiences, intercourse, or not using contraception increased in the 1990s and decreased in the 2000s, reflecting the changes in the abortion rate. However, as the abortion rate decreased, the intensity of sexual activity further increased among sexually experienced adolescents, suggesting that the fall in the abortion rate may be due to contraception and more effective sexual education. LEVEL OF EVIDENCE: : III. PMID- 19546763 TI - Human papillomavirus test after conization in predicting residual disease in subsequent hysterectomy specimens. AB - OBJECTIVE: To estimate the effectiveness of the human papillomavirus (HPV) test performed after conization in predicting residual disease in patients who subsequently underwent hysterectomy. METHODS: A total of 115 patients who underwent hysterectomy after conization caused by cervical intraepithelial neoplasia grade 3 (CIN 3) and microinvasive cervical cancer (IA1 cancer) were included in this prospective study. All patients underwent HPV testing with a liquid hybridization assay immediately before hysterectomy. Differences in sensitivity, specificity, and accuracy between resection margin and the HPV test in predicting residual disease in subsequent hysterectomy samples were estimated using the McNemar exact test. RESULTS: Univariable analysis showed that age, parity, menopausal status, glandular extension, and severity of disease were not predictive for residual disease, but positive resection margin and positive HPV tests were significant factors for predicting residual disease. These factors were also significant in a multivariable analysis (positive resection margin 45.5%, odds ratio [OR] 3.09, 95% confidence interval [CI] 1.19-8.03, P=.021; positive HPV test 57.6%, OR 11.05, 95% CI 4.01-30.49, P<.001). With resection margin, the sensitivity, specificity, and accuracy in predicting residual disease were 75%, 53%, and 61%, respectively, whereas, with the HPV test, these values were 85%, 67%, and 73%, respectively (P=.454, .080, and .044, respectively). Of patients with positive resection margins, 79% of HPV-negative patients had no residual disease. Of patients with negative resection margins, no HPV-negative patient had residual disease. CONCLUSION: The HPV test after conization was significantly more accurate than resection margin for predicting residual disease. The predictive value of resection margin for predicting residual disease was much improved when used in combination with the HPV test. Use of the HPV test is recommended for identifying patients for subsequent hysterectomy after conization for CIN 3 and IA1 cancer. LEVEL OF EVIDENCE: III. PMID- 19546764 TI - Parametrial involvement in radical hysterectomy specimens for women with early stage cervical cancer. AB - OBJECTIVE: To estimate the incidence of parametrial involvement and to evaluate factors associated with parametrial spread in women with early-stage cervical cancer and to identify a cohort of patients at low risk for parametrial spread who may benefit from less radical surgery. METHODS: We reviewed all patients who underwent radical hysterectomy and pelvic lymphadenectomy for invasive cervical cancer between 1990 and 2006. All women with squamous, adenocarcinoma, or adenosquamous disease, stage IA2-IB1, who underwent completed radical hysterectomy were included in the analysis. Normally distributed continuous variables were compared using Student's t-test for independent samples to analyze the outcome of positive or negative parametrial involvement. RESULTS: Three hundred fifty patients met the inclusion criteria. Overall, 27 women (7.7%) had parametrial involvement. The majority of specimens with parametrial involvement (52%) had tumor spread through direct microscopic extension. Patients with parametrial involvement were more likely to have a primary tumor size larger than 2 cm (larger than 2 cm: 14%, smaller than 2 cm: 4%, P=.001), higher histologic grade (grade 3: 12%, grades 1 and 2: 3%, P=.01), lymphovascular space invasion (positive: 12%, negative: 3%, P=.002), and metastasis to the pelvic lymph nodes (positive: 31%, negative: 4%, P<.001). One hundred twenty-five women (36%) had squamous, adenocarcinoma, or adenosquamous lesions, all grades, with primary tumor size 2 cm or smaller and no lymphovascular space invasion. In this group of patients, there was no pathologic evidence of parametrial involvement. CONCLUSION: We were able to retrospectively identify a cohort of women with early stage cervical cancer who were at very low risk for parametrial involvement. If prospective application of these findings confirms our results, less radical surgery-such as simple hysterectomy, simple trachelectomy, or conization-with pelvic lymphadenectomy may be a reasonable therapeutic option for women with primary tumors 2 cm or smaller and no lymphovascular space invasion. LEVEL OF EVIDENCE: III. PMID- 19546765 TI - Stage IIIA endometrial carcinoma: outcome and predictors of survival. AB - OBJECTIVE: To examine predictors of survival for stage IIIA endometrial cancer and to compare the outcome of stage IIIA neoplasms to that of endometrial tumors confined to the uterus. METHODS: : Women with stage IIIA endometrioid adenocarcinomas of the uterus treated between 1998 and 2004 were examined. Patients were stratified into two groups: positive cytology only (IIIA1) or ovarian or pelvic metastases (IIIA2). The clinical characteristics and outcome of stage IIIA1 and IIIA2 patients were compared. To further examine patients with positive cytology alone, women with IIIA1 tumors were compared with women with stage IB and IC tumors. Survival was examined using Cox proportional hazards models and the Kaplan-Meier method. RESULTS: : A total of 1,257 women, including 485 (39%) with IIIA1 and 772 (61%) with IIIA2 lesions, were identified. In a proportional hazards model incorporating known prognostic variables, there was no difference in survival (hazard ratio 1.06, 95% confidence interval [CI] 0.85 1.32) between IIIA1 and IIIA2 tumors. Five-year survival was 67% (95% CI 61-72%) for IIIA1 and 55% (95% CI 53-58%) for IIIA2 cancers. The results were unchanged when only women who underwent staging lymphadenectomy were included. Mortality was higher for IIIA1 tumors than stage IB and IC cancers. At 5 years, 90% (95% CI 89-90%) of patients with IB and 77% (95% CI 75-79%) with stage IC tumors were alive compared with 67% (95% CI 61-72%) of women with IIIA1 neoplasms. CONCLUSION: : Patients with stage IIIA1 tumors seem to have an outcome that is similar to that of women with serosal or adnexal metastases. The outcome of stage IIIA1 endometrial cancer is inferior to that of tumors confined to the corpus (stage IB and IC). LEVEL OF EVIDENCE: : II. PMID- 19546766 TI - Combination treatment for menstrual migraine and dysmenorrhea using sumatriptan naproxen: two randomized controlled trials. AB - OBJECTIVE: To evaluate the efficacy and tolerability of sumatriptan-naproxen during the mild pain phase of a single menstrual migraine attack associated with dysmenorrhea. METHODS: Two replicate randomized, multicenter, double-blind, placebo-controlled, trials of adults with menstrual migraine and dysmenorrhea were conducted. Participants treated their menstrual migraine attack during the mild pain phase (within 1 hour of onset) with sumatriptan 85 mg and naproxen sodium 500 mg in a single fixed-dose formulation (sumatriptan-naproxen) or placebo. The primary endpoint was 2-hour pain-free response. RESULTS: Sumatriptan naproxen was statistically superior to placebo in both studies (n=311, Study 1; n=310, Study 2) for 2-hour and, 2- to 24-hour sustained pain-free response, use of headache and menstrual rescue medications, and several nonpain menstrual symptom categories. Two-hour pain-free rates were Study 1, 42% compared with 23%, and Study 2, 52% compared with 22%, P<.001. Two- to 24-hour sustained pain-free rates were Study 1, 29% compared with 18%, P=.022; Study 2, 38% compared with 10%, P<.001. Headache and menstrual medication rates were Study 1, 37% compared with 53%, P=.005; Study 2, 31% compared with 69%, P<.001. Women treated with sumatriptan-naproxen continued to be pain free through 48 hours compared with placebo: Study 1, 26% compared with 17%, P=.040; Study 2, 28% compared with 8%, P<.001. No serious adverse events were reported in either study; nausea and dizziness were the most frequently reported adverse events. CONCLUSION: Sumatriptan-naproxen provided an effective pain-free response at 2 hours, which was maintained up to 48 hours in menstrual migraineurs with dysmenorrhea. Sumatriptan-naproxen was well-tolerated and resulted in decreased rescue medication use and relief of nonpainful menstrual symptoms. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00329459 and NCT00329355 LEVEL OF EVIDENCE: I. PMID- 19546767 TI - Factors associated with peripartum hysterectomy. AB - OBJECTIVE: To identify factors associated with peripartum hysterectomy performed within 30 days postpartum. METHODS: This was a population-based case-control study using Washington State birth certificate registry (1987-2006) linked to the Comprehensive Hospital Abstract Reporting System. Cases underwent hysterectomy within 30 days postpartum. Controls were frequency matched 4:1. Exposures included factors related to hemorrhage, delivery method, multiple gestations, and infection. Incidence rates of peripartum hysterectomy and maternal and neonatal morbidity and mortality were assessed. Adjusted odds ratios (aORs) by maternal age, parity, gestational age, year of birth, and mode of delivery and 95% confidence intervals (CIs) were computed. RESULTS: There were 896 hysterectomies. Incidence rates ranged from 0.25 in 1987 to 0.82 per 1,000 deliveries in 2006 (chi for trend, P<.001). Factors related to hemorrhage were strongly related to peripartum hysterectomy. Placenta previa (192 cases compared with 23 controls; aOR 7.9, 95% CI 4.1-15.0), abruptio placenta (71 compared with 55; aOR 3.2, 95% CI 1.8-5.8), and retained placenta (214 compared with 28; aOR 43.0, 95% CI 19.0 97.7) increased the risk of hysterectomy, as did uterine atony, uterine rupture, and thrombocytopenia. Having multiple gestations did not. As compared with vaginal delivery, vaginal delivery after cesarean (27 cases compared with 105 controls; aOR 1.9, 95% CI 1.2-3.0), primary cesarean (270 compared with 504; aOR 4.6, 95% CI 3.5-6.0), and repeat cesarean (296 compared with 231; aOR 7.9, 95% CI 5.8-10.7) increased the risk of peripartum hysterectomy. Among the 111 women who had hysterectomy on readmission (12.8% of cases), hemorrhage- and infection related factors were still strongly associated with peripartum hysterectomy. CONCLUSION: Incidence rates of peripartum hysterectomy are increasing over time. The most important risk factor for peripartum hysterectomy is hemorrhage, most notably caused by uterine rupture, retained placenta, and atony of uterus. LEVEL OF EVIDENCE: II. PMID- 19546768 TI - Diagnosing pulmonary embolism in pregnancy using computed-tomographic angiography or ventilation-perfusion. AB - OBJECTIVE: To estimate the rate of nondiagnosis for patients who initially undergo computed-tomographic angiography compared with those who undergo ventilation-perfusion imaging to diagnose pulmonary embolism in pregnancy. METHODS: This was a retrospective cohort study of all women consecutively evaluated from 2001-2006 for clinical suspicion of pulmonary embolism who were pregnant or 6 weeks postpartum and underwent at least computed-tomographic angiography or ventilation-perfusion scan. Charts were abstracted for history, clinical presentation, examination, imaging, and pregnancy and maternal outcomes. Women who underwent computed-tomographic angiography for initial diagnosis were compared with women who underwent ventilation-perfusion. Primary outcome was defined as a nondiagnostic study: nondiagnostic for pulmonary embolism in the computed-tomographic angiography group, or "low or intermediate probability" in the ventilation-perfusion group. Univariable, bivariable, and multivariable analyses were performed. RESULTS: Of 304 women with a clinical suspicion of pulmonary embolism, initial diagnosis was sought by computed-tomographic angiography in 108 (35.1%) and by ventilation-perfusion in 196 (64.9%) women. Women who underwent computed-tomographic angiography tended to have a slightly higher rate of nondiagnostic study (17.0% compared with 13.2%, P=.38). Examining the subgroup of women with a normal chest X-ray, computed-tomographic angiography was much more likely to yield a nondiagnostic result than ventilation-perfusion, even after adjusting for relevant confounding effects (30.0% compared with 5.6%, adjusted odds ratio 5.4, 95% confidence interval 1.4-20.1, P<.01). CONCLUSION: Pregnant or postpartum women with clinical suspicion of a pulmonary embolism and a normal chest X-ray are more likely to have a diagnostic study from a ventilation-perfusion scan compared with a computed-tomographic angiography. Evidence supports computed-tomographic angiography as a better initial test than ventilation-perfusion in patients with an abnormal chest X-ray. LEVEL OF EVIDENCE: II. PMID- 19546769 TI - Retention of chairs in obstetrics and gynecology: a comparison with other clinical departments. AB - OBJECTIVE: To compare the retention of chairs in academic obstetrics and gynecology with other core clinical departments. METHODS: Ongoing data were collected from each medical school for the Association of American Medical Colleges Faculty Roster between 1979 and 2007. Primary outcome measures included 5-year and 10-year retention rates and survival curves of first-time chairs. Comparisons were made between first-time chairs in obstetrics and gynecology and other core clinical departments: internal medicine, family medicine, pediatrics, psychiatry, and surgery. RESULTS: Five-year retention rates of obstetrics and gynecology chairs declined from 80% for those who began in 1979-1982 to 53% for those who began in 1998-2002. Ten-year retention in obstetrics and gynecology declined from 54% for those beginning in 1979-1982 to 26% for those beginning in 1993-1997. Other clinical departments experienced more stable 5-year and 10-year retention rates. Although substantially longer than other clinical departments in the 1979-1982 cohort, the median tenure of obstetrics and gynecology chairs who began in 1993-1997 was comparable with or less than that of other clinical departments. Discrete-time survival analysis revealed this decline in obstetrics and gynecology chair retention to be significant (P<.001) and more consistent than in other departments. CONCLUSION: Compared with other core clinical departments, retention of first-time chairs in obstetrics and gynecology declined more consistently from the highest to among the lowest. Chairs were inclined to not remain in office for a prolonged period. LEVEL OF EVIDENCE: II. PMID- 19546770 TI - The 2008 National Institute of Child Health and Human Development report on fetal heart rate monitoring. AB - Standardization of fetal heart rate (FHR) interpretation and management guidelines has been elusive, and no system is currently widely accepted in the United States. The recently summarized 2008 Eunice Kennedy Shriver National Institute of Child Health and Human Development workshop proposed a three-tier system of interpretation of FHR patterns, but left management recommendations to the professional associations. The middle tier, called indeterminate Category II, which contains the variant FHR patterns seen most frequently, is vast and heterogeneous. We propose that this category can be subcategorized at least tentatively, based on evidence available from previously published studies. Such subcategorization will allow the organizations proposing management recommendations to more readily set up guidelines for graded interventions and clinical responses to the spectrum of FHR patterns, with the aim of minimizing fetal acidemia without excessive obstetric intervention. Such management algorithms will need to be tested by appropriately designed clinical studies. PMID- 19546771 TI - Human papillomavirus infection: a concise review of natural history. AB - Persistent infection with oncogenic human papillomavirus (HPV), leading to precancerous lesions and potentially cervical cancer, is a serious health burden. The natural history of infection is one that enables the virus to remain immunoevasive within the cervical epithelium and persist for decades. Many studies have provided important information on why some women clear infection and why others do not. The infectious process of HPV may be affected by many cofactors, adding to the complexity of disease development. Vaccination against oncogenic HPV along with cervical screening are two methods that have been developed to provide protection and eliminate the overall burden of disease in women. This article highlights the natural history of oncogenic HPV infection of the cervix and the limitations that currently exist on the literature. PMID- 19546772 TI - The legacy of Ralph Reis. PMID- 19546773 TI - Trauma in pregnancy. AB - Acute traumatic injury during pregnancy is a significant contributor to maternal and fetal morbidity and mortality in the United States. Motor vehicle accidents are the leading cause of injury-related maternal death, followed by violence and assault. Lack of seat belts or other restraints increases the risks of both maternal and fetal morbidity and mortality. The American College of Obstetricians and Gynecologists recommends proper seat belt use by all pregnant women and screening for domestic abuse. Maternal injury and death from physical abuse is prevalent, and in some communities, homicide is a major cause of pregnancy associated maternal death. Blunt trauma most often occurs as a result of motor vehicle accidents, whereas penetrating trauma results from gunshots or stabbings. Blunt trauma to the abdomen increases the risk for placental abruption, and direct fetal injury is more likely with penetrating trauma. Management strategies in acute maternal trauma must focus on a thorough assessment of the mother. A coordinated team effort that includes the obstetrician is essential to ensure optimal maternal and fetal outcomes. Imaging studies should not be delayed because of concerns of fetal radiation exposure, because the risk is minimal with usual imaging procedures, especially in mid-to-late pregnancy. The obstetrician should serve in a consultative role if nonobstetric surgical care is required and must also be prepared to intervene on behalf of the mother and the fetus if trauma care is compromised by the pregnancy. Perimortem cesarean delivery should be considered early in the resuscitation of a pregnant trauma victim, especially when fetal viability is a concern. Once the mother is stabilized in the emergency setting, she should be transported for appropriate maternal and fetal observation until both mother and fetus are clear of danger. It is essential that the clinician and staff maintain thorough and accurate documentation and recording of the chronology of events, the maternal and fetal assessment, and the management and outcome of the pregnancy. PMID- 19546775 TI - Medications in pregnancy and lactation. PMID- 19546776 TI - Medications in pregnancy and lactation. PMID- 19546779 TI - Robotic gynecologic surgery. PMID- 19546780 TI - Using vasopressin for myomectomy. PMID- 19546782 TI - Time from decision to incision for cesarean deliveries at a community hospital. PMID- 19546783 TI - Tocolytic therapy: a meta-analysis and decision analysis. PMID- 19546785 TI - Influence of intraoperative capsule rupture on outcomes in stage I epithelial ovarian cancer. PMID- 19546789 TI - Preventing excessive weight gain during pregnancy through dietary and lifestyle counseling: a randomized controlled trial. PMID- 19546790 TI - Cervical cancers after human papillomavirus vaccination. PMID- 19546791 TI - Cervical cancers after human papillomavirus vaccination. PMID- 19546792 TI - One-day compared with 7-day nitrofurantoin for asymptomatic bacteriuria in pregnancy: a randomized controlled trial. PMID- 19546793 TI - Eligibility criteria in venous thromboembolism, myocardial infarction, and stroke among transdermal contraceptive system users. PMID- 19546798 TI - ACOG Practice Bulletin No. 106: Intrapartum fetal heart rate monitoring: nomenclature, interpretation, and general management principles. PMID- 19546799 TI - ACOG Committee Opinion No. 437: Community involvement and volunteerism. PMID- 19546800 TI - The visible ear simulator: a public PC application for GPU-accelerated haptic 3D simulation of ear surgery based on the visible ear data. AB - BACKGROUND: Existing virtual simulators for middle ear surgery are based on 3 dimensional (3D) models from computed tomographic or magnetic resonance imaging data in which image quality is limited by the lack of detail (maximum, approximately 50 voxels/mm3), natural color, and texture of the source material.Virtual training often requires the purchase of a program, a customized computer, and expensive peripherals dedicated exclusively to this purpose. MATERIALS AND METHODS: The Visible Ear freeware library of digital images from a fresh-frozen human temporal bone was segmented, and real-time volume rendered as a 3D model of high-fidelity, true color, and great anatomic detail and realism of the surgically relevant structures. A haptic drilling model was developed for surgical interaction with the 3D model. RESULTS: Realistic visualization in high fidelity (approximately 125 voxels/mm3) and true color, 2D, or optional anaglyph stereoscopic 3D was achieved on a standard Core 2 Duo personal computer with a GeForce 8,800 GTX graphics card, and surgical interaction was provided through a relatively inexpensive (approximately $2,500) Phantom Omni haptic 3D pointing device. CONCLUSION: This prototype is published for download (approximately 120 MB) as freeware at http://www.alexandra.dk/ves/index.htm.With increasing personal computer performance, future versions may include enhanced resolution (up to 8,000 voxels/mm3) and realistic interaction with deformable soft tissue components such as skin, tympanic membrane, dura, and cholesteatomas-features some of which are not possible with computed tomographic-/magnetic resonance imaging-based systems. PMID- 19546801 TI - Venous thromboembolism in lymphoma: how effectively are we treating patients? AB - INTRODUCTION: Patients with solid tumors and venous thromboembolic episodes (VTE) have a high risk of recurrence and bleeding during oral anticoagulant treatment. However, we are unaware of studies expressly evaluating such risks in patients with lymphoma. Therefore, we conducted a retrospective study to determine the frequency of such complications during treatment of lymphoma patients who develop VTE. METHODS: Charts of patients with histologically proven non-Hodgkin lymphoma and Hodgkin lymphoma were retrospectively reviewed and patients with their first acute symptomatic VTE episode were identified (49 non-Hodgkin lymphoma, 8 Hodgkin lymphoma). Recurrence of VTE episodes and major and minor bleeding during treatment with warfarin or low molecular weight heparin (LMWH) were recorded. RESULTS: All 57 patients were initially treated with high-dose-adjusted intravenous heparin or body-weight-adjusted LMWH. Forty-six patients were started on oral warfarin and 11 patients continued LMWH. Recurrent VTE episodes occurred in 14 of 46 patients on warfarin therapy, whereas major bleeding was documented in 6 of 46 patients, and minor bleeding in 9 of 46 patients. Recurrent VTE episodes occurred in 1 of 11 patients treated with LMWH, whereas major bleeding occurred in 0 of 11 and minor bleeding in 3 of 11 patients. CONCLUSIONS: Lymphoma patients treated with warfarin experienced a 30.4% rate of recurrent thrombosis and 13% major bleeding. During this treatment most (65%), but not all, bleeding and thrombotic complications occurred with an international normalized ratio within the therapeutic range. The percentage of serious complications (recurrent VTE and major bleeding) during warfarin use was 44.5%, and the death rate was 6.5%, compared with 9% and 0%, respectively, during use of LMWH. PMID- 19546802 TI - Accuracy of breast cancer axillary lymph node treatment plans based on 2 dimensional imaging: what we should know before interpreting 2-dimensional treatment-planning era studies. AB - OBJECTIVE: To investigate axillary 2-dimensional treatment planning accuracy. METHODS: Computed tomography (CT) simulation data for 16 breast cancer cases taken after level I-II axillary dissection were analyzed. An additional 6 patients underwent CT simulation using the historical 90-degree position (HP), and the standard-bore CT position (CT-P). Two physicians identified the lateral and medial borders of the coracoid process (CCP) on digitally reconstructed radiography (DRR). The DRR-identified x coordinates were compared with the CT measured x coordinates. x coordinates differences between the most medial surgical clip and the borders of the CCP as identified on CT were analyzed. Fields were designed to cover various amounts of the axilla, and treatment plans were generated to compare doses to the most medial surgical clip. RESULTS: In 11 and 6 cases for each physician, respectively (lateral border), and in all cases for both physicians (medial border), the DRR identification of the CCP was medial to that on CT. In 9 and 8 cases, the most medial surgical clip was lateral to the medial and lateral borders of the CCP, respectively. In all data sets, the average difference was larger in the HP compared with CT position. The number of patients who received more than 90% of the prescribed dose when using the plans with the mid humeral head border, lateral border of the CCP, and medial border of the CCP were as follows: 6, 1, and 0, respectively. CONCLUSIONS: When using 2 dimensional treatment planning, the dose to the undissected axilla can vary depending on the anatomic landmark used to define the lateral border of the axillary field. This may account for outcome differences found in older radiotherapy studies. PMID- 19546804 TI - Continuous spinal anesthesia. AB - Continuous spinal anesthesia (CSA) is an underutilized technique in modern anesthesia practice. Compared with other techniques of neuraxial anesthesia, CSA allows incremental dosing of an intrathecal local anesthetic for an indefinite duration, whereas traditional single-shot spinal anesthesia usually involves larger doses, a finite, unpredictable duration, and greater potential for detrimental hemodynamic effects including hypotension, and epidural anesthesia via a catheter may produce lesser motor block and suboptimal anesthesia in sacral nerve root distributions. This review compares CSA with other anesthetic techniques and also describes the history of CSA, its clinical applications, concerns regarding neurotoxicity, and other pharmacologic implications of its use. CSA has seen a waxing and waning of its popularity in clinical practice since its initial description in 1907. After case reports of cauda equina syndrome were reported with the use of spinal microcatheters for CSA, these microcatheters were withdrawn from clinical practice in the United States but continued to be used in Europe with no further neurologic sequelae. Because only large-bore catheters may be used in the United States, CSA is usually reserved for elderly patients out of concern for the risk of postdural puncture headache in younger patients. However, even in younger patients, sometimes the unique clinical benefits and hemodynamic stability involved in CSA outweigh concerns regarding postdural puncture headache. Clinical scenarios in which CSA may be of particular benefit include patients with severe aortic stenosis undergoing lower extremity surgery and obstetric patients with complex heart disease. CSA is an underutilized technique in modern anesthesia practice. Perhaps more accurately termed fractional spinal anesthesia, CSA involves intermittent dosing of local anesthetic solution via an intrathecal catheter. Where traditional spinal anesthesia involves a single injection with a somewhat unpredictable spread and duration of effect, CSA allows titration of the block level to the patient's needs, permits a spinal block of indefinite duration, and can provide greater hemodynamic stability than single-injection spinal anesthesia. PMID- 19546803 TI - Economic analysis of radiation therapy oncology group 97-14: multiple versus single fraction radiation treatment of patients with bone metastases. AB - INTRODUCTION: Radiation Therapy Oncology Group 97-14 concluded that a single fraction of radiation was as effective in relieving pain as multiple fractions in the treatment of patients with bone metastases. A statistically significant higher retreatment rate, however, was noted in patients undergoing a single fraction treatment. The purpose of the analysis was to determine whether multiple fraction treatment is cost-effective in treating patients with bone metastasis, by preventing further retreatment. METHODS AND MATERIAL: A Markov model was used to evaluate the cost-effectiveness of 30 Gy in 10 fractions in comparison with 8 Gy in 1 fraction. Transition probabilities, cost, and utilities were obtained from the clinical trial. Costs and outcomes were not discounted because of the short time line for the study. RESULTS: The expected mean cost and quality adjusted survival in months for patients receiving 8 Gy in 1 fraction and 30 Gy in 10 fractions was 998 US dollars and 7.26 months and 2316 US dollars and 9.53 months, respectively. The incremental cost-effectiveness ratio was 6973 US dollars/quality-adjusted life year. The results were sensitive to the utility of the posttreatment state for both single and multiple fraction treatments. CONCLUSION: Single fraction treatment was the less expensive treatment in the treatment of patients with bone metastasis treated on Radiation Therapy Oncology Group 97-14. PMID- 19546805 TI - Severe preeclampsia: anesthetic implications of the disease and its management. AB - Preeclampsia is a significant, multifactorial, multiorgan disease affecting 6%-8% of all pregnancies in the United States and is the third leading cause of maternal mortality. As such, it is incumbent upon any anesthesia provider involved in obstetric cases to be familiar with the varied manifestations of the disease, management goals from an obstetric standpoint, and the implications for provision of anesthesia in this patient group. Despite improvements in the diagnosis and management of preeclampsia, severe complications can occur in both the mother and the fetus. A systematic approach to the anesthetic evaluation is therefore necessary to reduce morbidity and mortality and to improve outcomes. The potential pitfalls of general anesthesia, including failed intubation, in these complicated patients make regional anesthesia the preferred choice in many cases. Recent studies have shown that spinal anesthesia is often appropriate for preeclamptic patients, even in severe cases. Nevertheless, it is important to be aware of the potential contraindications to neuraxial anesthesia and to prepare for the possibility of encountering a difficult airway. PMID- 19546806 TI - Achieving electric-acoustic benefit with a modulated tone. AB - OBJECTIVE: When either real or simulated electric stimulation from a cochlear implant (CI) is combined with low-frequency acoustic stimulation (electric acoustic stimulation [EAS]), speech intelligibility in noise can improve dramatically. We recently showed that a similar benefit to intelligibility can be observed in simulation when the low-frequency acoustic stimulation (low-pass target speech) is replaced with a tone that is modulated both in frequency with the fundamental frequency (F0) of the target talker and in amplitude with the amplitude envelope of the low-pass target speech (). The goal of the current experiment was to examine the benefit of the modulated tone to intelligibility in CI patients. DESIGN: Eight CI users who had some residual acoustic hearing either in the implanted ear, the unimplanted ear, or both ears participated in this study. Target speech was combined with either multitalker babble or a single competing talker and presented to the implant. Stimulation to the acoustic region consisted of no signal, target speech, or a tone that was modulated in frequency to track the changes in the target talker's F0 and in amplitude to track the amplitude envelope of target speech low-pass filtered at 500 Hz. RESULTS: All patients showed improvements in intelligibility over electric-only stimulation when either the tone or target speech was presented acoustically. The average improvement in intelligibility was 46 percentage points due to the tone and 55 percentage points due to target speech. CONCLUSIONS: The results demonstrate that a tone carrying F0 and amplitude envelope cues of target speech can provide significant benefit to CI users and may lead to new technologies that could offer EAS benefit to many patients who would not benefit from current EAS approaches. PMID- 19546807 TI - Audiovisual deficits in older adults with hearing loss: biological evidence. AB - OBJECTIVE: To examine the impact of hearing loss (HL) on audiovisual (AV) processing in the aging population. We hypothesized that age-related HL would have a pervasive effect on sensory processing, extending beyond the auditory domain. Specifically, we predicted that decreased auditory input to the neural system, in the form of HL over time, would have deleterious effects on multisensory mechanisms. DESIGN: This study compared AV processing between older adults with normal hearing (N = 12) and older adults with mild to moderate sensorineural HL (N = 12). To do this, we recorded cortical evoked potentials that were elicited by watching and listening to recordings of a speaker saying the syllable "bi." Stimuli were presented in three conditions: when hearing the syllable "bi" (auditory), when viewing a person say "bi" (visual), and when seeing and hearing the syllables simultaneously (AV). Presentation level of the auditory stimulus was set to +30 dB SL for each listener to equalize auditory input across groups. RESULTS: In the AV condition, the normal-hearing group showed a clear and consistent decrease in P1 and N1 latencies as well as a reduction in P1 amplitude compared with the sum of the unimodal components (auditory + visual). These integration effects were absent or less consistent in HL participants. CONCLUSIONS: Despite controlling for auditory sensation level, visual influence on auditory processing was significantly less pronounced in HL individuals compared with controls, indicating diminished AV integration in this population. These results demonstrate that HL has a deleterious effect on how older adults combine what they see and hear. Although auditory amplification vastly improves the communication abilities in most hearing impaired individuals, the associated atrophy of multisensory mechanisms may contribute to a patient's difficulty in everyday settings. Our findings and related studies emphasize the potential value of multimodal tasks and stimuli in the assessment and rehabilitation of hearing impairments. PMID- 19546809 TI - Application of dual-genome oligonucleotide array-based comparative genomic hybridization to the molecular diagnosis of mitochondrial DNA deletion and depletion syndromes. AB - PURPOSE: Mitochondrial disorders constitute a group of clinically and genetically heterogeneous diseases for which molecular diagnosis has been a challenge. The current procedures for diagnosis of mitochondrial DNA deletion and depletion syndromes based on Southern analysis and quantitative polymerase chain reaction are particularly inefficient for determining important parameters of deletion endpoints and percent heteroplasmy. We have developed an improved approach for routine analyses of these disorders in a clinical laboratory. METHODS: A custom designed oligonucleotide array-based comparative genomic hybridization platform was developed to provide both tiled coverage of the entire 16.6-kb mitochondrial genome and high-density coverage of nuclear genes involved in mitochondrial biogenesis and function, for quick evaluation of mitochondrial DNA deletion and depletion. RESULTS: For initial validation, the performance of this array was characterized in 20 samples with known mitochondrial DNA deletions and 12 with apparent depletions. All previously known deletions were clearly detected and the break points were correctly identified by the oligonucleotide array-based comparative genomic hybridization, within the limits of resolution of the array. The extent of mitochondrial DNA depletion and the percentage of deletion heteroplasmy were estimated using an automated computational approach that gave results comparable to previous methods. Conclusions from subsequent application of this approach with >300 new clinical samples have been in 100% concordance with those from standard methods. Finally, for one sample, we were able to identify an intragenic deletion in a nuclear gene that was responsible for the observed mitochondrial DNA depletion. CONCLUSION: We conclude that this custom array is capable of reliably detecting mitochondrial DNA deletion with elucidation of the deletion break points and the percentage of heteroplasmy. In addition, simultaneous detection of the copy number changes in both nuclear and mitochondrial genomes makes this dual genome array of tremendous value in the diagnoses of mitochondrial DNA depletion syndromes. PMID- 19546808 TI - Sensory processing of linguistic pitch as reflected by the mismatch negativity. AB - OBJECTIVE: To assess the extent to which acoustic and phonetic change-detection processes contribute to the mismatch negativity (MMN) to linguistic pitch contours. DESIGN: MMN was elicited from Mandarin and English speakers using a passive oddball paradigm. Two oddball conditions were constructed. In one condition (T1/T2i), the Mandarin high-level tone (T1) was compared with a convex high-rising tone (inverted T2, henceforth referred to as T2i) that occurs as a contextual variant of T1 in running speech. In the other (T2/T2i), the concave high-rising tone (T2) was compared with T2i. Phonetically, T1/T2i represents a within-category contrast for native speakers, whereas T2/T2i represents a between category contrast. The between-category pair (T2/T2i), however, is more similar acoustically than the within-category pair (T1/T2i). In an attention-demanding behavioral paradigm, the same speakers also performed an auditory discrimination task to determine the perceptual distinctiveness of the two tonal pairs. RESULTS: Results revealed that the Chinese group, relative to the English, showed larger MMN responses and earlier peak latencies for both conditions, indicating experience-dependent enhancement in representing linguistically relevant pitch contours. At attentive stages of processing, however, the Chinese group was less accurate than the English in discriminating the within-category contrast (T1 T2i). CONCLUSIONS: These findings demonstrate that experience-dependent neural effects at early preattentive stages of processing may be driven primarily by acoustic features of pitch contours that occur in natural speech. At attentive stages of processing, perception is strongly influenced by tonal categories and their relations to one another. The MMN is a useful index for examining long-term plasticity to linguistically relevant acoustic features. PMID- 19546810 TI - Technical standards and guidelines for myotonic dystrophy type 1 testing. AB - Myotonic dystrophy type 1 is an autosomal dominant multisystem condition. Myotonic dystrophy type 1 is the result of an unstable CTG expansion in the 3' untranslated region of the myotonic dystrophy protein kinase gene. The age of onset and the severity of the phenotype are roughly correlated with the size of the CTG expansion. The combination of Southern transfer and polymerase chain reaction provides an accurate means of identifying patients affected by myotonic dystrophy type 1. This document follows the outline format of the general Standards and Guidelines for Clinical Genetics Laboratories. It is designed to be a checklist for genetic testing professionals who are already familiar with the disease and the methods of analysis. PMID- 19546811 TI - Maraviroc concentrates in the cervicovaginal fluid and vaginal tissue of HIV negative women. AB - OBJECTIVE: To compare single- and multiple-dose maraviroc exposures in cervicovaginal fluid (CVF) and vaginal tissue (VT) with blood plasma (BP) and quantify maraviroc protein binding in CVF. DESIGN: Open-label pharmacokinetic study. METHODS: In 12 HIV-negative women, 7 paired CVF and BP samples were collected over 12 hours after 1 maraviroc dose. Subjects then received maraviroc twice daily for 7 days. After the last dose, subjects underwent CVF and BP sampling as on day 1, with additional sampling during terminal elimination. VT biopsies were obtained at steady state. RESULTS: Day 1 and day 7 median maraviroc CVF AUCtau were 1.9- and 2.7-fold higher, respectively, than BP. On day 1, 6 of 12 subjects had detectable maraviroc CVF concentrations within 1 hour; 12 of 12 were detectable within 2 hours, and all exceeded the protein-free IC90. On day 7, maraviroc CVF protein binding was 7.6% and the VT AUCtau was 1.9-fold higher than BP. Maraviroc CVF concentrations 72 hours after dose and BP concentrations 12 hours after dose were similar. CONCLUSIONS: Higher maraviroc exposure in the female genital tract provides a pharmacologic basis for further evaluation of chemokine receptor 5 antagonists in HIV infection prophylaxis. This is the first study to report antiretroviral VT concentrations, CVF protein binding, and CVF terminal elimination. PMID- 19546812 TI - CD4+ response and subsequent risk of death among patients on antiretroviral therapy in Lusaka, Zambia. AB - INTRODUCTION: Where virologic monitoring is not routinely available, immunologic criteria are commonly used to determine treatment failure while on antiretroviral therapy (ART). However, few have studied CD4+ response and its relationship to subsequent clinical outcomes in a programmatic setting. METHODS: We analyzed cohort data from Zambia to investigate whether 6- and 12-month CD4+ response after ART initiation was associated with later mortality. We used Cox proportional hazards models that accounted for different strata of baseline CD4 counts and adjusted for age, sex, clinical stage, tuberculosis coinfection, baseline hemoglobin, initial ART regimen, and adherence behavior. RESULTS: We analyzed data from 2 cohorts, from 6 months onward (n = 24,366; median follow-up = 467 days, interquartile range 222-791) and from 12 months onward (n = 17,920; median follow-up = 423 days, interquartile range 191-689). In the post-6-month analysis, hazard for death was significantly higher when absolute CD4+ response was <100 cells per microliter [adjusted hazard ratio (AHR) = 2.25, 95% confidence interval (CI): 1.91 to 2.64], relative response was <10% above baseline (AHR = 2.60, 95% CI: 2.12 to 3.19), and absolute CD4+ count was <100 per microliter (AHR = 2.79, 95% CI: 2.26 to 3.45). In the post-12 month analysis, mortality was associated with rise in absolute CD4+ cell count <200 per microliter (AHR = 2.41, 95% CI: 1.83 to 3.17), relative rise in CD4+ cell count of <10% above baseline (AHR = 3.41, 95% CI: 2.51 to 4.64), and absolute CD4+ count at 12 months <100 per microliter (AHR = 4.11, 95% CI: 2.96 to 5.68). CONCLUSION: Commonly used definitions for immunologic treatment failure are associated with elevated mortality risk among patients on ART. PMID- 19546813 TI - Time trends and regional differences in the prevalence of HIV infection among women attending antenatal clinics in 2 provinces in Cameroon. AB - BACKGROUND: HIV prevalence time trends vary in sub-Saharan African countries. In the present study, we studied time trends and regional differences in the prevalence of HIV infection among women attending antenatal care clinics (ANC) in 7 sites located in 2 provinces in Cameroon. METHODOLOGY: As part of ANC, 16,626 women consented to HIV testing from 2000 to 2006. Sociodemographic and risk factor information was collected during the initial 3 years of the study. This information was aggregated within sites and used as site-level covariate in multilevel logistic regression analysis. RESULTS: HIV prevalence decreased significantly in women younger than 20 years from 13% in 2000 to 5% in 2006. Age specific prevalence varied among the sites, with a peak prevalence occurring more often at a higher age in 2004-2006 versus 2000-2003, suggesting a reduction of HIV incidence over time. There was a substantial heterogeneity across sites in HIV prevalence, which was lower in sites where women had earlier sexual debut and were less well educated. CONCLUSIONS: ANC surveillance indicates a decreasing trend in HIV prevalence in the studied sites in Cameroon. Cultural differences might have accounted for the heterogeneity of HIV infection observed across sites, which call for tailored interventions. PMID- 19546814 TI - S-propargyl-cysteine protects both adult rat hearts and neonatal cardiomyocytes from ischemia/hypoxia injury: the contribution of the hydrogen sulfide-mediated pathway. AB - In this study, we determined the cardioprotective effects of S-propargyl-cysteine (SPRC), a structural analog of S-allylcysteine (SAC), using in vivo models of acute myocardial infarction (MI) and in vitro hypoxic cardiomyocytes models. MI was created in rats by ligating the left anterior descending coronary artery. Plasma enzymes levels and cystathionine-gamma-lyase (CSE) activities were determined. Primary cultures of newborn rats' cardiomyocytes were injured by hypoxia for 6 h. Cell viabilities were measured with the thiazolyl blue assay. RT PCR and western blot analysis revealed the expression of CSE in both models. The protective effects of SPRC were associated with an observed reduction in infarct size (20.8 +/- 2.4% vs. 36.0 +/- 1.3%), decreased plasma enzymes levels and reduced malondialdehyde levels when compared to the MI vehicle group (P < 0.05); cardiac function was also improved. SPRC increased CSE activity and plasma H2S concentration by 1.6-fold and 1.3-fold, respectively, in MI rats. Decreased cell viability (64.5 +/- 5.4%) in hypoxic cardiomyocytes could be rescued with use of SPRC (81.0 +/- 3.1%). Similarly, mRNA and protein expression of CSE were upregulated in the SPRC group. Treatment with the CSE inhibitor propargylglycine abolished the protective effects of SPRC. Our study provides novel evidence that SPRC is protective in myocardial infarctions via a H2S-related pathway. PMID- 19546815 TI - TRC4186, a novel AGE-breaker, improves diabetic cardiomyopathy and nephropathy in Ob-ZSF1 model of type 2 diabetes. AB - Advanced glycation end products (AGEs) contribute significantly to diabetic complications, both macro- and microvascular. TRC4186 is an AGE-breaker that has been evaluated in vitro and in vivo and shown to reduce AGE burden. The aim of this study was to determine the effect of TRC4186 on diabetic cardiomyopathy and nephropathy in obese Zucker spontaneously hypertensive fatty rats (Ob-ZSF1), an animal model of diabetes with progressive cardiac and renal dysfunction. Ob-ZSF1 rats loaded with 0.5% salt were treated with TRC4186, 9 or 27 mg/kg twice daily intraperitoneally or vehicle control and monitored telemetrically throughout the study. Cardiac function was assessed terminally by Millar catheter. Markers of cardiac and renal dysfunction were measured and changes evaluated histopathologically. TRC4186 at 27 mg/kg prevented rise in blood pressure (BP) and also improved cardiac output (CO) secondary to better diastolic relaxation as well as systolic emptying in association with the reduction in afterload. At 9 mg/kg, CO was improved by compensatory increase in pre-load however afterload reduction was not adequate to allow efficient systolic emptying. Brain natriuretic peptide (BNP) and interleukin-6 (IL-6) expression was reduced with treatment. Deterioration in renal function was retarded as evident from albumin to creatinine ratio and renal histopathology. TRC4186, an AGE-breaker, clearly preserved cardiac function and reduced the severity of renal dysfunction in Ob ZSF1, an animal model with persistent severe hyperglycemia leading to diabetic heart failure and renal failure. PMID- 19546816 TI - Reversal of postprandial endothelial dysfunction by cyclooxygenase inhibition in healthy volunteers. AB - The aim of this study was to evaluate the role of cyclooxygenase (COX) in venous vascular reactivity changes after an oral lipid overload (OLO). Venous endothelial function (dorsal hand vein technique) was evaluated in fasting, 30 minutes after COX inhibition (aspirin-fasting), 2 to 4 hours after an OLO (1000 kcal, 58% fat), and again after COX inhibition (aspirin-OLO, 600 mg/200 mL water) in 10 healthy adults (age, 28.1 +/- 1.3 years; body mass index, 22.3 +/- 0.6 kg/m). Fasting, 2- to 4-hour post-OLO, and 60-minute postaspirin plasma glucose, insulin, and lipids were also evaluated. The OLO increased triglycerides and insulin, reduced low-density lipoprotein and high-density lipoprotein, but glycemia and total cholesterol remained unchanged. There were no metabolic differences between OLO and aspirin-OLO. In fasting, aspirin reduced acetylcholine-induced venodilation (107.0% +/- 14% versus 57.3% +/- 11%; P < 0.001). Vascular reactivity was blunted after the OLO (phenylephrine dose: 0.3 +/ 0.2 fasting versus 1.9 +/- 0.8 nmol/min after OLO; P < 0.001) and was partially corrected by aspirin (0.4 +/- 0.2; P < 0.001). Similar changes were observed in maximum venodilation after acetylcholine (107.0% +/- 14% fasting versus 60.4% +/- 9% after OLO, P < 0.001; aspirin-OLO: 95.9% +/- 6%; P < 0.001). The responses to sodium nitroprusside remained unchanged during the study. We conclude that the OLO reduction in the endothelium-dependent venoconstriction and venodilation is partially the result of the action of COX. PMID- 19546819 TI - Outcome for patients with malignant pleural mesothelioma referred for Trimodality therapy in Western Australia. AB - INTRODUCTION: Trimodality therapy (TMT), consisting of extrapleural pneumonectomy (EPP), preoperative or postoperative combination chemotherapy, and high-dose hemithoracic radiotherapy, is the only therapy reported to achieve long-term survival in selected patients with malignant pleural mesothelioma (MPM). Thus, TMT was introduced as an option for such patients in Western Australia in 2004. However, TMT has never been compared with non-TMT therapy in the same patient population, thereby introducing a potential for selection bias. METHOD: We performed a retrospective review of all patients referred for TMT consisting of EPP, adjuvant chemotherapy, and hemithoracic radiotherapy at a quaternary referral institution. Patient eligibility for referral for TMT was based on patients' tolerability for pneumonectomy, epithelioid subtype, and computed tomography and positron emission tomography scanning indicating operable disease, with the exclusion of extrapleural lymphadenopathy and metastatic disease (clinical stage T1-3N0-1M0). Eligible patients consenting to TMT also underwent a surgical staging procedure (bilateral thoracoscopy, mediastinoscopy, and laparoscopy) to confirm eligibility before EPP. RESULTS: Thirty-six patients have been referred for TMT since 2004, and there has been a median of 27 months follow up; of 31 patients having surgical staging, eight were ineligible for EPP and one declined EPP. Of the 22 planned for EPP, 18 underwent EPP and four had unresectable disease at surgery. There was one death in hospital six days post EPP and another death postdischarge and 28 days post-EPP (30-day mortality 11%); 15 of 16 EPP survivors received adjuvant chemotherapy and 14 completed adjuvant radiotherapy. Pathologic analysis of the 18 resected EPP specimens revealed N2 disease in seven patients (39%) and nonepithelioid subtype in six patients (33%). Local recurrence did not occur among EPP survivors; however, 56% (9 of 16 patients) developed distant recurrence. Median and 1-year survival did not differ between the 18 EPP patients and 18 non-EPP patients (20.4 versus 20.7 months and 76 versus 78%, respectively; p = NS). DISCUSSION: In this case series, we could not demonstrate a survival benefit for patients in the EPP group compared with that in the non-EPP group. After surgical staging, 26% of patients were ineligible for TMT. Thus, surgical staging is essential before proceeding with EPP. Despite aggressive imaging and surgical staging, 39% of patients will have N2 disease and 18% will have unresectable disease at operation. Although complete locoregional control was achieved with TMT, distant recurrence affected most EPP survivors despite careful patient selection and a high rate of completion of adjuvant therapy. We conclude that TMT for operable epithelioid MPM requires further assessment in randomized controlled trials. PMID- 19546818 TI - Gemcitabine in brief versus prolonged low-dose infusion, both combined with cisplatin, for advanced non-small cell lung cancer: a randomized phase II clinical trial. AB - OBJECTIVE: Gemcitabine in low dose in prolonged infusion is a treatment with documented activity against a variety of tumors. We here report the first randomized trial to compare standard brief and low-dose prolonged infusion of gemcitabine. PATIENTS AND METHODS: Eligible patients had non-small cell lung cancer in stage IIIB (wet) or IV, Karnofsky performance status 100 to 70 (Eastern Cooperative Oncology Group 0-2), measurable disease, were chemonaive and fulfilled the standard criteria for chemotherapy. In arm A (standard treatment), gemcitabine was given at 1250 mg/m(2) in 20 to 30 minutes and in arm B (prolonged infusion) at 250 mg/m(2) in 6 hours infusion. All patients received gemcitabine on days 1 and 8 and cisplatin at 75 mg/m(2) on day 2 of a 3-week cycle for four cycles, followed by two cycles of gemcitabine as monotherapy. RESULTS: A total of 249 patients (188 men and 61 women, median age 58 years) were randomized between arm A (125 patients) and arm B (124 patients). Adenocarcinoma (53.9%) was the predominant histologic type; 92% of patients were in stage IV. The two groups were balanced for prognostic factors; however, group A had fewer patients with significant weight loss and no patient with lung cancer as a second malignancy or after radiotherapy for brain metastases. Grade 3 or greater toxicity was rare: anemia in 0.8 and 3.2%, neutropenia in 21.6 and 22.6%, thrombocytopenia in 0 and 1.6%, and nausea/vomiting in 4 and 8.1% for arms A and B, respectively. Alopecia was seen in 54.5% of patients in arm B, as compared with 9.7% in arm A. No patient died of treatment-related toxicity. During cycle 5, 47.7% of patients in arm A and 60.7% in arm B reported improved well-being, as compared with the status before chemotherapy. Patients in arm A had no complete remission, 32.8% partial responses, 48% minimal responses or stable disease, 13.6% progressions, and 5.6% were not evaluable. For arm B, the corresponding figures are as follows: complete remission 0.8%, partial responses 46% (for overall response rate of 46.8%), minimal responses or stable disease 36.3%, progression 12.1%, and not evaluable 4.8%. Median progression-free survival was 5.5 and 6.0 months, median overall survival was 10.1 and 10.0 months, and 1-year survival was 46.6 and 41.1% for arms A and B, respectively. For the 71 patients with squamous carcinoma, arm B seems superior to arm A, as seen by the higher overall response rate (51.3 versus 35.5%), longer median progression-free survival (6.2 versus 4.9 months), and longer median survival (11.3 versus 8.5 months). However, because of the small number of patients, these differences did not reach the level of statistical significance. CONCLUSION: In the treatment of advanced non-small cell lung cancer, gemcitabine in low dose in prolonged infusion in combination with cisplatin has low toxicity and has activity comparable with gemcitabine in higher dose in standard brief infusion. Low-dose gemcitabine may be preferred for incurable cancer among economically deprivileged patients. In addition, apparent superior activity against squamous carcinoma opens new perspectives and deserves further research. PMID- 19546817 TI - Pretreatment quality of life is an independent prognostic factor for overall survival in patients with advanced stage non-small cell lung cancer. AB - HYPOTHESIS: We conducted this pooled analysis to assess the prognostic value of pretreatment Quality of Life (QOL) assessments on overall survival (OS) in advanced non-small cell lung cancer (NSCLC). METHODS: Four hundred twenty patients with advanced NSCLC (stages IIIB with pleural effusion and IV) from six North Central Cancer Treatment Group trials were included in this study. QOL assessments included the single-item Uniscale (355 patients), Lung Cancer Symptom Scale (217 patients), and Functional Assessment of Cancer Therapy-Lung (197 patients). QOL scores were transformed to a 0 to 100 scale with higher scores representing better status and categorized using the sample median or clinically deficient score (CDS, 50). Cox proportional hazards models stratified by study were used to evaluate the prognostic importance of QOL on OS alone and in the presence of other prognostic factors such as performance status, age, gender, body mass index, and laboratory parameters. RESULTS: Pretreatment QOL accessed by Uniscale was significantly associated with OS univariately (p < 0.0001). Uniscale (p < 0.0001; hazard ratio = 1.6 for the sample median and 2.0 for the CDS categorization) and body mass index were the only significant predictors of OS multivariately. The median survival of patients who had a Uniscale score less than or equal to the CDS (50 group; and 7.8 versus 13 months for the less than or equal to sample median (83 group, respectively. The Lung Cancer Symptom Scale and the Functional Assessment of Cancer Therapy-Lung total scores were not significant predictors of OS. CONCLUSIONS: Pretreatment QOL measured by Uniscale is a significant and an independent prognostic factor for OS, and QOL should be routinely integrated as a stratification factor in advanced NSCLC trials. PMID- 19546820 TI - Gefitinib in combination with gemcitabine and carboplatin in never smokers with non-small cell lung carcinoma: a retrospective analysis. AB - INTRODUCTION: Randomized placebo-controlled phase III trials failed to show a survival benefit with the addition of gefitinib to platinum-based combination chemotherapy as first-line therapy in unselected patients with advanced non-small cell lung cancer (NSCLC). We conducted a retrospective analysis of the outcome in never smokers with advanced NSCLC who received gemcitabine-carboplatin-gefitinib (GCI) as first-line therapy and compared these patients with a historical control group who received gemcitabine-carboplatin (GC) alone in our center. METHODS: Never-smoker patients with chemonaive stage IIIB or IV NSCLC were treated with GCI. These patients were compared with a historical control group of never smokers who had been treated with GC alone as the first-line therapy. RESULTS: A total of 80 patients were reviewed: 51 patients were treated with GCI and 29 with GC. Most patients were women, and adenocarcinoma was the most common histologic subtype. The response rate for patients in the GCI group was 62.7% (95% confidence interval [CI] = 48.08-75.87), which was higher than that of the GC group, 27.6% (95% CI = 12.73-47.24). The GCI group showed a significant improvement in progression-free survival compared with the GC group (hazard ratio of 0.19, 95% CI = 0.105-0.351, p < 0.001). The median overall survival for the patients on GCI was 20.5 months compared 14.1 months (p = 0.05) for patients on GC. CONCLUSION: The addition of gefitinib to first-line chemotherapy improved progression-free survival and overall survival when used as a first-line therapy in never smokers with advanced NSCLC in this retrospective study. A prospective randomized phase III study is needed to confirm this finding. PMID- 19546821 TI - Survey and biological insights of pemetrexed-related therapeutic improvement in mesothelioma: The Nancy Centre of Biological Resources' Mesothelioma Cohort. AB - INTRODUCTION: We report a survey of mesothelioma survival rates with insights into the survival benefit because of pemetrexed. We also studied a potential link between specific single nucleotide polymorphisms of transcobalamin II (TCII) gene and susceptibility to both asbestos and pemetrexed. METHODS: Clinical and occupational data from 287 consecutive mesothelioma patients were collected from the north-east region of France (1989-2007). Blood or paired tumoral and normal samples were collected from the last 210 French patients to study the TCII single nucleotide polymorphisms at the codon 259 (quantitative polymerase chain reaction). Results were compared with those obtained from a group of 263 French control healthy subjects and to a group of 91 German mesothelioma patients. Patients' characteristics and genotypes results were statistically analyzed for significant correlations. RESULTS: The mean overall patient's survival was 18.19 +/- 21.07 months. Pemetrexed increased the patients' survival by 50% (21.81 versus 16.99 months). The TCII allele Proline (Pro) was overrepresented into the mesothelioma cohort when compared with the controls (35 versus 19.77%). This also concerned German patients. The alleles Pro and Proline Arginine (ProArg) were more frequent among patients exposed to asbestos (p = 0.005, p < 0.001, respectively). The allele ProArg was associated with the longest survival while under pemetrexed (p = 0.007). No difference was found in the genotypes of patients untreated with pemetrexed. CONCLUSIONS: Pemetrexed treatment is related to a survival increase in mesothelioma patients. The allele Pro seems overrepresented in mesothelioma patients. Those having the allele ProArg present a better outcome under pemetrexed. PMID- 19546822 TI - Blunt cerebrovascular trauma: distal internal carotid dissection. PMID- 19546823 TI - Vitamin D and fat: the chicken versus the egg. PMID- 19546824 TI - Symptoms reported by women in midlife: menopausal transition or aging? AB - OBJECTIVE: The aim of this study was to determine which symptoms commonly reported by women at midlife are associated with the menopausal transition, after adjusting for aging, life events, sociodemographics, and lifestyle factors. METHODS: Middle-aged women participating in the Australian Longitudinal Study on Women's Health between 1996 (survey 1, ages 45-50 y) and 2007 (survey 5) were included in the analyses if natural menopause status could be determined at any survey (n = 8,649 of 13,716 participants). Natural menopause status was determined from reported menstruation patterns. A survival function describing age at menopause was computed. Logistic regression models for repeated measures were used to estimate the association between menopausal stage and symptom prevalence. RESULTS: There were 6,814 (79%) women who reached natural menopause before 2007. The median age at menopause was 52 years. Compared with the premenopausal phase, menopause was associated with hot flushes (odds ratio, 8.6 [95% CI, 7.5-9.9]), night sweats (odds ratio, 5.5 [95% CI, 4.8-6.3]), and, to a lesser extent, stiff or painful joints (odds ratio, 1.6 [95% CI, 1.4-1.8]), difficulty sleeping (odds ratio, 1.4 [95% CI, 1.2-1.6]), and poor/fair self-rated health (odds ratio, 1.6 [95% CI, 1.3-1.9]), after controlling for confounders. Prevalence of some symptoms was still raised more than 7 years after menopause. Headaches/migraines were negatively associated with aging, and urinary incontinence was positively associated with aging. CONCLUSIONS: Treatment such as hormone therapy should be targeted to vasomotor symptoms, which are most strongly associated with menopause rather than to less specific symptoms related to aging per se. PMID- 19546825 TI - Bazedoxifene effects on the reproductive tract in postmenopausal women at risk for osteoporosis. AB - OBJECTIVE: The aim of this study was to examine the endometrial, ovarian, and breast safety of bazedoxifene, a novel selective estrogen-receptor modulator, in postmenopausal women at risk for osteoporosis. METHODS: Healthy postmenopausal women (N = 1,583; mean age, 57.6 y) with lumbar spine or femoral neck bone mineral density T scores between -1 and -2.5 and/or other clinical risk factors for osteoporosis were enrolled in a 24-month, phase 3, randomized, double-blind, placebo- and active-controlled trial. They received daily treatment with bazedoxifene 10, 20, or 40 mg; placebo; or raloxifene 60 mg. Reproductive safety assessments included periodic transvaginal ultrasound measurements of endometrial thickness, ovarian volume, and presence of ovarian cysts; periodic endometrial biopsies; and adverse event reporting. RESULTS: Bazedoxifene was not associated with a significant change from baseline in mean endometrial thickness at month 24. The percentage of participants with a change from baseline in endometrial thickness or endometrial thickness greater than 5 mm at month 24 was similar among groups. There was no consensus diagnosis of endometrial hyperplasia or malignancy in the bazedoxifene or raloxifene groups; the rates of other histologic findings, including endometrial polyps, were low (<5%) and similar among groups. No significant between-group differences were found in the change from baseline in ovarian volume, number or size of ovarian cysts, or incidence of ovarian cancer. Reports of breast pain (<4%) and breast cancer (<1%) were low and evenly distributed among groups. CONCLUSION: A favorable endometrial, ovarian, and breast safety profile was found after 2 years of treatment with bazedoxifene in healthy, recently postmenopausal women at risk for osteoporosis. PMID- 19546826 TI - Relief of vasomotor symptoms with the tissue-selective estrogen complex containing bazedoxifene/conjugated estrogens: a randomized, controlled trial. AB - OBJECTIVE: The aim of this study was to assess the safety and efficacy of bazedoxifene (BZA)/conjugated estrogens (CE) treating moderate to severe vasomotor symptoms in the Selective Estrogen Menopause and Response to Therapy 2 trial. METHODS: This was an outpatient, multicenter, double-blind, randomized, placebo-controlled, phase 3 study conducted in the United States. Healthy postmenopausal women (N = 332; aged 40-65 y) with moderate to severe hot flushes (>or=7/d or 50/wk) were randomized to BZA 20 mg/CE 0.45 mg, BZA 20 mg/CE 0.625 mg, or placebo once daily for 12 weeks. Changes from baseline in the average daily number of moderate and severe hot flushes and daily severity score were assessed at weeks 4 and 12; adverse events were recorded. RESULTS: BZA/CE significantly reduced the number and severity of hot flushes at weeks 4 and 12 (P < 0.001). At week 12, BZA 20 mg/CE 0.45 mg and BZA 20 mg/CE 0.625 mg reduced hot flushes from baseline by 74% (10.3 hot flushes [baseline] vs 2.8 [week 12]) and 80% (10.4 vs 2.4), respectively, compared with 51% (10.5 vs 5.4) for placebo. More participants at week 12 had at least a 75% decrease in hot flushes with BZA 20 mg/CE 0.45 mg (61%) and BZA 20 mg/CE 0.625 mg (73%) versus placebo (27%; P < 0.001). The safety profile was similar between BZA/CE and placebo, and no unexpected safety findings were reported. CONCLUSIONS: BZA 20 mg paired with CE 0.45 or 0.625 mg is effective, with short-term safety, for treating vasomotor symptoms in postmenopausal women. PMID- 19546827 TI - Nogo in the Mammalian cochlea. AB - HYPOTHESIS: Different members of the Nogo system are expressed in the mammalian cochlea. BACKGROUND: The protein Nogo has gained a lot of attention during the last couple of years because it inhibits neurite outgrowth in the adult central nervous system. In contrast to the central nervous system, very little is known regarding the expression and possible function of the Nogo system within the inner ear. METHODS: Using reverse-transcriptase-polymerase chain reaction and immunohistochemistry, we analyzed for the expression of members of the Nogo system within the cochlea. In addition, we determined hearing levels of Nogo A knockout and wild-type mice with auditory brainstem response audiometry. RESULTS: In this study, we demonstrate the expression of Nogo A, B, C, and of Nogo receptor mRNA in the organ of Corti, spiral ganglion, and stria vascularis. Immunohistochemistry revealed that Nogo A and Nogo receptor localize to the spiral ganglion neurons. Interestingly, Nogo A expression was also observed in the outer and inner hair cells of the organ of Corti. As revealed by light microscopy, deletion of Nogo A does not alter cochlear microanatomy. We have assessed hearing levels in 10-month old wild-type and Nogo A knockout mice, and thereby, we could not detect any differences between these 2 groups. CONCLUSION: Different members of the Nogo family are expressed in the mammalian cochlea. Deletion of Nogo A does not alter cochlea microanatomy or hearing levels compared with wild-type mice. PMID- 19546828 TI - Unilateral sensorineural hearing loss in children: the importance of temporal bone computed tomography and audiometric follow-up. AB - OBJECTIVES: The first objective of this study was to identify rates, types, and laterality of clinically relevant inner ear malformations in children with unilateral sensorineural hearing loss (USNHL). The second objective was to assess the change of the ipsilesional and contralesional hearing thresholds of the patients with USNHL and the association between hearing change with time and the findings on high-resolution temporal bone computed tomography (TBCT). STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: A total of 322 children diagnosed with USNHL on initial audiometry were included. INTERVENTIONS: Otologic examination, pure-tone audiometry or auditory brainstem response, and high-resolution TBCT. MAIN OUTCOME MEASURES: Radiologic findings demonstrating clinically relevant bony or soft tissue malformations of the inner ear. Audiologic findings of change in thresholds. RESULTS: In a series of 322 consecutively investigated children with USNHL, 93 TBCT scans (28.9%) were identified as abnormal. The abnormal CT findings included cochleovestibular malformations (49 cases; 52.7%), vestibular malformations (27 cases; 29.0%), and malformations of vestibular or cochlear aqueducts (17 cases; 18.3%). Of these abnormal CT findings, 18 cases (19.4%) showed bilateral malformations. Incomplete partition type II was the most common type of malformation (28 cases), followed by narrow internal auditory canal (23 cases) and subsequently followed by enlarged vestibular aqueduct syndrome (17 cases). Of 244 patients, 77 (31.6%) in the profound USNHL group accompanied malformations, which was significantly higher than 23.8% (15 of 78) in the nonprofound (mild to severe) USNHL group (p = 0.036). Of 115 patients who were regularly followed up for more than 6 months, 7 (6.1%) had hearing decrement (including 3 cases of bilateral decrement). Of 85 patients in the normal TBCT group, 4 (4.7%) revealed hearing decrement, and considering there were only 27 patients with nonprofound USNHL in this group, the rate of decrement was as much as 14.8% (4 of 27). Of 24 patients in the unilateral malformation group, 1 patient (4.2%) showed worsening of hearing, and 2 of 6 (33.3%) patients showed worsening of hearing in the bilateral malformation group. CONCLUSION: Based on the results, we propose all children with USNHL should have a TBCT scan because management, including genetic counseling and prognostic predictions of these cases, may be significantly influenced by the CT outcome. Moreover, the non-negligible rate of progressive nature of ipsilateral and contralateral SNHL in normal-looking TBCT group and in groups with inner ear malformations mandates longitudinal audiologic assessments for both ears PMID- 19546829 TI - The long-term effects of modified electrode surfaces and intracochlear corticosteroids on postoperative impedances in cochlear implant patients. AB - OBJECTIVE: The objective of this study was to investigate the long-term effect of intraoperative application of steroid suspension and coating of the electrode contacts with a thin film of iridium oxide on the intracochlear impedance development after cochlear implantation and on the impedance difference before and after stimulation. DESIGN: Time-dependent development of intracochlear impedances was investigated in 4 different groups of adult patients up to 4 years after implantation. Additionally, during rehabilitation period just after first fitting, impedances before and after stimulation were measured as to investigate the influence of electrical stimulation on the impedances. Results from standard Nucleus 24 Contour (control), standard Nucleus 24 Contour with intraoperative application of steroids, iridium-coated Nucleus 24 Contour, and iridium-coated Nucleus 24 Contour with intraoperative application of steroids were compared. RESULTS: Steroid application reduced impedances significantly throughout the observation period of up to 4 years after implantation. Iridium oxide coating had no effect. Differences between the groups were mainly found on the basal and middle parts of the cochlea, but not close to the tip of the array, also indicating that postoperative fibrous tissue growth is stronger in the basal region of the cochlea. Group mean values of the stimulation effect were not influenced by the different treatments. Nevertheless, only in both steroid treated groups a correlation between the impedance before stimulation and the stimulation effect was found. CONCLUSION: Although the differences between control and steroid-treated groups decrease with time, single intraoperative intracochlear steroid deposition was proven to lower postoperative impedances during first 3 to 4 years after implantation probably because of reduction of fibrous tissue growth. PMID- 19546830 TI - Laryngeal mask airway use in otologic surgery. AB - OBJECTIVE: The purpose of this study was to evaluate the safety and efficacy of the laryngeal mask airway (LMA) compared with oral endotracheal tube (ETT) in general anesthesia in patients who underwent otologic surgery. STUDY DESIGN: Retrospective review. SETTING: Military tertiary care teaching hospital. PATIENTS: A retrospective chart review study was conducted on 484 patients who underwent otologic surgery at a large military tertiary care teaching hospital from January 2002 to August 2005. A review of otologic surgical cases using the LMA (n = 167) was compared to a similar cohort managed with an oral ETT (n = 317). MAIN OUTCOME MEASURES: The patient's age, weight, indication for surgery, comorbid conditions, American Society of Anesthesiologists classification, duration of surgery, and adverse events were recorded. Specific data reviewed included airway complications, intraoperative times, anesthetic medications, postoperative nausea and vomiting, and time spent in the postanesthesia care unit. RESULTS: There were no major airway complications in either group requiring an emergency airway. A significant decreased use of neuromuscular blocking agents was noted in the LMA group. Statistical analysis of variances was performed using 2-sample Wilcoxon rank-sum (Mann-Whitney U) test and Fisher's exact test. The time in the operating room to the start of surgery and the time from completion of surgery to patient exiting the operating room were shorter for the LMA group than for the ETT group. There were no statistical differences in postanesthesia care unit time or postoperative nausea and vomiting. CONCLUSION: The LMA offers a safe alternative in children and adults to endotracheal intubation with no observed increased risk of airway complications in patients undergoing otologic surgery. PMID- 19546831 TI - Genetics of otosclerosis. AB - OBJECTIVES: Otosclerosis is a major cause of acquired hearing loss in adult life affecting exclusively the human temporal bone. Until recently, the etiopathogenesis of otosclerosis was still a matter of debate. Genetic research, however, has evolved enormously the last years and unveiled important clues regarding the cause of otosclerosis. The objective of this article is to review the genetics of otosclerosis with special attention for the links to the bone homeostasis of the otic capsule. DATA SOURCES: A detailed literature study was performed focusing on the recent genetic findings in otosclerosis and the special bone turnover of the otic capsule. A PubMed search and own research data were used to bring the relevant information for this review together. CONCLUSION: Unlike all other bones in the human skeleton, the otic capsule undergoes very little remodeling after development, possibly due to local inner ear factors. Otosclerosis is a process of pathologic increased bone turnover in the otic capsule, which in most cases leads to stapes fixation, resulting in a conductive hearing loss. Although environmental factors such as estrogens, fluoride, and viral infection have been implicated, it is clear that genetic factors play a significant role in the manifestation of otosclerosis. From a genetic viewpoint, otosclerosis is considered to be a complex disease with rare autosomal dominant forms caused by a single gene. Already, 7 monogenic loci have been published, but none of the genes involved have been identified. For the complex form of otosclerosis, caused by an interaction between genetic and environmental factors, the first susceptibility genes were identified by case-control association studies. All 3 replicated genes, TGFB1, BMP2, and BMP4, are a part of the transforming growth factor-beta1 pathway. Data from both genetic association studies and gene expression analysis of otosclerotic bone showed that the TGF beta1 pathway is most likely an important factor in the pathogenesis of otosclerosis. PMID- 19546832 TI - Outcomes in nontumor adults fitted with the auditory brainstem implant: 10 years' experience. AB - OBJECTIVES: Recently, open-set speech recognition performance has been observed with electric stimulation of the brainstem in some nontumor (NT) patients. These outcomes require that we reevaluate the criteria for patient selection and the rationale for expanding the application for the auditory brainstem implant (ABI) to NT adult patients with profound hearing loss. This study presents results from up to 10 years in adults, with analysis based on cause. MATERIALS AND METHODS: In our Department, from April 1997 to September 2007, ABIs have been implanted in 112 patients (83 adults and 29 children) with tumor (T) and NT disorders. Of the 112 patients, 15 have previously had a cochlear implant elsewhere with no sound detection. This article presents speech recognition results from the 80 patients (32 neurofibromatosis type 2 and 48 NT) who had achieved at least 1-year follow up. The retrosigmoid approach was used in all NT patients, and the retrosigmoid transmeatal approach was used in all T patients. RESULTS: All patients had a functioning implantation, and no complications were observed during the operation, activation, or long-term use of the ABI. At the most recent follow-up, NT adults scored from 10 to 100% in open-set speech perception tests (average, 59%), and T patients scored from 5 to 31% (average, 10%). The differences between these results are statistically significant (p = 0.0007). The best performance was observed in patients who lost their nerve VIII from head trauma or severe ossification. Lowest performance (although still highly beneficial to the patient) was observed in patients with neurologic disorders, neuropathy, and cochlear malformations. CONCLUSION: Our experience clearly indicates that the ABI is an effective tool for hearing rehabilitation in patients with profound hearing loss who cannot be fitted with cochlear implants. PMID- 19546833 TI - Genetic determinants of hearing loss associated with vestibular schwannomas. AB - HYPOTHESIS: The severity of hearing loss (HL) associated with vestibular schwannomas (VSs) is influenced by genes expressed by the VSs. BACKGROUND: Hearing loss is the most common presenting symptoms in patients with VSs, yet its pathophysiology remains elusive. Previous studies have suggested that VSs cause HL not only by inducing degeneration of the auditory nerve by compression but also by promoting degeneration of the inner ear. This study aimed to determine whether there is a molecular basis for differences in HL associated with VSs. METHODS: Surgical specimens of VSs were collected from 13 patients and were divided into a group associated with good (word recognition >70% and pure-tone average < or =30 dB) or poor hearing. Whole-genome expression profiling of VSs was performed with the Affymetrix GeneChip Human X3P Array. The expression of select genes was validated using real-time quantitative reverse transcription polymerase chain reaction and immunohistochemistry. Because of a small sample size, exact nonparametric tests were used to assess the association between good versus poor hearing and specific histological features of the tumors and patient demographics. RESULTS: Using gene set enrichment analysis, the chromosomal region 3q27 was found to be significantly different between the 2 groups of tumors. This region includes peroxisomal biogenesis factor 5-like gene, which was underexpressed in VSs with poor hearing. The expression of 3 other genes from different chromosomes was significantly different between the 2 groups: RAD54B, prostate-specific membrane antigen-like, and carcinoembryonic antigen. CONCLUSION: This study identified several molecular alterations in VSs stratified by hearing. These alterations may determine the severity of HL associated with VSs and may represent potential therapeutic targets to prevent or reduce HL in theses patients. PMID- 19546834 TI - Increase of in vivo antitumoral activity by CD40L (CD154) gene transfer into pancreatic tumor cell-dendritic cell hybrids. AB - OBJECTIVES: Fusion of dendritic cells (DC) with tumor cells is an approach in immunotherapy combining antigenicity and capacity of antigen presentation to activate T cells for the induction of tumor-specific cytotoxic immunity. Although there have been reports of clinical benefit, response rates have been limited and further improvements are warranted. METHODS: We used murine DC and a novel protocol for an effective fusion of those cells with the murine pancreatic cell line Panc02. RESULTS: We observed 2 events: only moderate in vitro and in vivo cytotoxicity of tumor cell/DC hybrids and a down-regulation of costimulatory molecules on fused cells. Therefore, we transfected tumor cell/DC hybrids with an adenovirus expressing CD154 to improve DC activation and generating antitumor immune response without the need of CD4 T cells. High CD154 expression could be obtained by transfection of DC and Panc02 cells prior fusion. Furthermore, vaccination with CD154-transfected tumor cell/DC hybrid led to a significantly increased induction of cytotoxic T cells in vitro and to an improved antitumoral effect in an orthotopic in vivo mouse model. CONCLUSIONS: CD154-transfected tumor cell/DC hybrids are a promising approach to increase the efficiency of antitumoral response. PMID- 19546836 TI - The usefulness of perfusion-weighted magnetic resonance imaging in advanced pancreatic cancer. AB - OBJECTIVES: Perfusion-weighted magnetic resonance imaging (MRI) can detect the changes of signal intensity in tumors. We evaluated the prognostic value of perfusion-weighted MRI in patients with advanced pancreatic cancer (PC). METHODS: Perfusion-weighted MRI was performed before treatment on 27 consecutive patients with advanced PC. The American Joint Committee on Cancer (AJCC) stages of patients were as follows (8, stage III; 19, stage IV). Imaging acquisition was continually repeated with echo planar sequence every 2 seconds for 2 minutes after a bolus injection of gadolinium. We made a time intensity curve of PC and calculated the signal ratio (SR) on perfusion-weighted imaging. We assessed the relation between SR and clinical factors including tumor stage, lymph node metastasis, liver metastasis, and so on. Patients were divided into low and high SR group and compared SR with the overall survival. RESULTS: All cases showed transient decreases signal intensity (SR, 6.9-55.7%). These patients were classified into 2 groups at cutoff median SR of 22.0% The high SR group significantly correlated with the higher stage (P=0.03) and the presence of lymph node metastasis (P=0.04). The high SR group had significantly shorter overall survival (P=0.04). CONCLUSIONS: Perfusion-weighted MRI may predict the survival in advanced PC patients. PMID- 19546835 TI - Serum REG4 level is a predictive biomarker for the response to preoperative chemoradiotherapy in patients with pancreatic cancer. AB - OBJECTIVES: Preoperative chemoradiotherapy is one of the key strategies for the improvement of survival in pancreatic cancer; however, no method to predict the response has yet been established. The aim of this study was to prospectively evaluate the predictive value of REG4, a new member of the regenerating (REG) islet-derived family of proteins. METHODS: Stably REG4-expressing cells were established from a pancreatic cancer cell line and exposed in vitro to gamma-ray or gemcitabine to investigate the relevance of REG4 to the resistance to chemotherapy or radiotherapy. In 23 patients with resectable pancreatic cancer, the serum concentration of REG4 was measured before preoperative chemoradiotherapy, and the histologic response was evaluated after the surgery. RESULTS: A 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and fluorescence activated cell scanning (FACS) revealed that REG4 overexpressing cells were resistant to gamma-radiation but showed a modest resistance to gemcitabine. The patients with a higher REG4 level, but not carcinoembryonic antigen or CA-19-9, showed an unfavorable histologic response to chemoradiotherapy (Spearman, rho = 0.439, P = 0.039). The patients showing a higher REG4 level experienced local recurrence postoperatively. CONCLUSIONS: These data demonstrated in vitro and in vivo that REG4 protein overexpression was associated with an unfavorable response to preoperative chemoradiotherapy. REG4 can clinically be used as a predictive biomarker. PMID- 19546837 TI - Differences in pancreatic immunohistochemical staining profiles of TGF-beta1, MMP 2, and TIMP-2 between autoimmune and alcoholic chronic pancreatitis. AB - OBJECTIVES: Tumor growth factor beta (TGF-beta) is an immunosuppressive cytokine and has been implicated in a variety of disease processes, including those in autoimmune disease. Tumor growth factor beta is also involved in fibrosis by regulating matrix metalloproteinases (MMPs) and the tissue inhibitor of MP (TIMP). The purpose of this study was to compare the expression patterns of TGF beta1, MMP-2, and TIMP-2 between autoimmune chronic pancreatitis (AIP) and alcoholic chronic pancreatitis (ACP) by immunohistochemical staining of pancreatic tissue specimens. METHODS: Pancreatic tissue specimens were obtained from 16 of 57 patients who had a diagnosis of AIP at the Asan Medical Center. Pancreatic tissue specimens of ACP were obtained from 10 patients who were surgically treated. Immunohistochemical staining was performed with antibodies specific for TGF-beta1, MMP-2, and TIMP-2. RESULTS: The degree of immunohistochemical staining for TGF-beta1 was significantly weaker in AIP than in ACP in the pancreatic ductal epithelial and mononuclear cells (P = 0.029 and P = 0.018, respectively). CONCLUSIONS: This finding suggests that there may be a defect in the function of regulatory T (Treg) cells, which normally prevents autoimmune disease progression via a suppressor mechanism. Further studies are needed to identify the type of regulatory T cell involved in this process. PMID- 19546838 TI - Antithrombin III prevents cerulein-induced acute pancreatitis in rats. AB - OBJECTIVES: Systemic inflammatory mediators, including the protein high-mobility group box 1 (HMGB1), play an important role in the development of acute pancreatitis. Anticoagulants, such as antithrombin III (AT III), inhibit inflammation resulting from various causes, but their mechanism of action is not well understood. Because acute pancreatitis is a severe inflammatory disease, we hypothesized that AT III would inhibit inflammation and prevent cerulein-induced acute pancreatitis. METHODS: Experimental animals received or were saline injected with a bolus of 250 IU/kg of AT III followed by intraperitoneal injections of 50 mg/kg of cerulein. Levels of cytokines (interleukin 6 and tumor necrosis factor alpha), nitric oxide (NO), and HMGB1 were measured in serum and pancreatic tissue at regular intervals for 12 hours after the cerulein injection. RESULTS: Pancreas histopathology and wet-dry ratio significantly improved in the AT III-injected (250 IU/kg) animals compared with the saline-injected rats. Serum and pancreas HMGB1 levels decreased over time in AT III-treated animals. Antithrombin III also decreased cytokine, NO, and HMGB1 levels during cerulein induced inflammation. As a result, AT III ameliorated the pathologic pancreas in the rat model of cerulein-induced acute pancreatitis. CONCLUSIONS: Antithrombin III treatment inhibited the secretion of cytokines, NO, and HMGB1 and prevented cerulein-induced acute pancreatitis in the rat model. PMID- 19546839 TI - Protocol for management of imported pediatric malaria decreases time to medication administration. AB - BACKGROUND: A malaria management protocol was developed and implemented at a tertiary care children's hospital in September 1999. We retrospectively evaluated children admitted with malaria 10-years preimplementation and 7-years postimplementation to determine the impact the protocol had on management and time delay to appropriate antimalarial therapy. METHODS: This before and after study compared all admissions with the discharge diagnosis of malaria in the study period. Retrospective chart review was used to determine the time from emergency department (ED) registration to administration of antimalarial treatment. Other outcomes measured included mortality, length of hospital stay, and intensive care unit admission. RESULTS: Fifty-eight admissions were identified during the defined period, most of which were due to Plasmodium falciparum[r] malaria. Thirty-one (53.4%) cases were before implementation of the protocol. Children were more likely to receive appropriate investigations to assess for possible severe malaria before transfer from the ED to the ward after protocol implementation (18% vs. 63%, P = 0.005). Analysis of index cases of malaria, excluding patients diagnosed after the diagnosis of a sibling, showed there was a significant reduction in time to medication administration (8 vs. 5.5 hours, P = 0.036). CONCLUSION: After broad-based implementation of a malaria treatment protocol in a pediatric hospital, children received more thorough investigations, were more likely to receive therapy before leaving the ED and had a shorter delay before receiving appropriate antimalarial therapy. PMID- 19546840 TI - The impact of pneumococcal conjugate vaccine on rates of myringotomy with ventilation tube insertion in Australia. AB - BACKGROUND: In randomized controlled trials and postmarketing studies the heptavalent pneumococcal conjugate vaccine (7vPCV) has been shown to reduce myringotomy with ventilation tube insertion (MVTI) procedures in a 4-dose schedule. In Australia, a 3-dose schedule at 2, 4, and 6 months of age is routinely used in non-Indigenous children. Our aim was to determine if a reduction in MVTI comparable to that documented in the United States occurred in Australia despite the absence of the booster dose. METHODS: All episodes of MVTI in Australia from July 1998 to June 2007 among children aged 0 mmHg. A second phase (with progressively longer CSF draining) further defined the diagnosis. A ventriculo-peritoneal shunt was then placed and the nerve function returned to normal within few days. Third cranial nerve dysfunction as a predominant sign of hydrocephalus is very rare and may raise doubts as to the real significance of the imaging findings of enlarged ventricles. In this ground, ICP monitoring is a safe and helpful diagnostic tool that can afford a more accurate evaluation and proper treatment. The supposed mechanism of 3(rd) nerve dysfunction was bending/stretching of the nerve. PMID- 19546848 TI - Co-existing thyrotropin secreting pituitary adenoma and low grade glioma: clinical considerations and literature review. AB - The authors report an extremely rare co-occurrence of a thyrotropin secreting pituitary adenoma and a low grade glioma. A 35-year old patient was initially misdiagnosed and treated as a case of primary hyperthyroidism till he presented with visual symptoms and seizures. On evaluation he was also found to have a right frontal glioma. They discuss the presentation, clinical and diagnostic implications, treatment options and possible molecular pathways which help us understand the molecular biology of such tumor associations. They review literature of 12 previous indexed reports of pituitary tumors associated with various grades of gliomas. Though there are 3 previous reports of functioning pituitary tumors and gliomas, this is the first report of a thyrotropin secreting pituitary adenoma and an associated low grade glioma. PMID- 19546850 TI - Joint reanalysis of 29 correlated SNPs supports the role of PCLO/Piccolo as a causal risk factor for major depressive disorder. PMID- 19546851 TI - Hydrogen sulfide impairs keratinocyte cell growth and adhesion inhibiting mitogen activated protein kinase signaling. AB - The effects of exogenous hydrogen sulfide (H2S) on normal skin-derived immortalized human keratinocytes have been investigated in detail. We show in vitro that exogenous hydrogen sulfide reduces clonal growth, cell proliferation and cell adhesion of human keratinocytes. H(2)S, in fact, decreases the frequency of the putative keratinocyte stem cell subpopulation in culture, consequently affecting clonal growth, and impairs cell proliferation and adhesion of mature cells. As a mechanistic explanation of these effects, we show at the molecular level that (i) H2S reduces the Raf/MAPK kinase/ERK signaling pathway; (ii) the reduced adhesion of sulfur-treated cells is associated to the downregulation of the expression of beta4, alpha2 and alpha6 integrins that are necessary to promote cell adhesion as well as anti-apoptotic and proliferative signaling in normal keratinocytes. One specific interest of the effects of sulfurs on keratinocytes derives from the potential applications of the results, as sulfur is able to penetrate the skin and a sulfur-rich balneotherapy has been known for long to be effective in the treatment of psoriasis. Thus, the relevance of our findings to the pathophysiology of psoriasis was tested in vivo by treating psoriatic lesions with sulfurs at a concentration comparable to that most commonly found in sulfurous natural springs. In agreement with the in vitro observations, the immunohistochemical analysis of patient biopsies showed a specific downregulation of ERK activation levels, the key molecular event in the sulfur-induced effects on keratinocytes. PMID- 19546852 TI - Endothelial Notch signaling is upregulated in human brain arteriovenous malformations and a mouse model of the disease. AB - Brain arteriovenous malformations (BAVMs) can cause lethal hemorrhagic stroke and have no effective treatment. The cellular and molecular basis for this disease is largely unknown. We have previously shown that expression of constitutively active Notch4 receptor in the endothelium elicits and maintains the hallmarks of BAVM in mice, thus establishing a mouse model of the disease. Our work suggested that Notch pathway could be a critical molecular mediator of BAVM pathogenesis. Here, we investigated the hypothesis that upregulated Notch activation contributes to the pathogenesis of human BAVM. We examined the expression of the canonical Notch downstream target Hes1 in the endothelium of human BAVMs by immunofluorescence, and showed increased levels relative to either autopsy or surgical biopsy controls. We then analyzed receptor activity using an antibody to the activated form of the Notch1 receptor, and found increased levels of activity. These findings suggest that Notch activation may promote the development and even maintenance of BAVM. We also detected increases in Hes1 and activated Notch1 expression in our mouse model of BAVM induced by constitutively active Notch4, demonstrating molecular similarity between the mouse model and the human disease. Our work suggests that activation of Notch signaling is an important molecular candidate in BAVM pathogenesis and further validates that our animal model provides a platform to study the progression as well as the regression of the disease. PMID- 19546853 TI - Slow progressive conduction and contraction defects in loss of Nkx2-5 mice after cardiomyocyte terminal differentiation. AB - Mutations in homeoprotein NKX2-5 are linked to human congenital heart disease, resulting in various cardiac anomalies, as well as in postnatal progressive conduction defects and occasional left ventricular dysfunction; yet the function of Nkx2-5 in the postnatal period is largely unexplored. In the heart, the majority of cardiomyocytes are believed to complete cell-cycle withdrawal shortly after birth, which is generally accompanied by a re-organization of chromatin structure shown in other tissues. We reasoned that the effects of the loss of Nkx2-5 in mice may be different after cell-cycle withdrawal compared with those of the perinatal loss of Nkx2-5, which results in rapid conduction and contraction defects within 4 days after the deletion of Nkx2-5 alleles (Circ Res. 2008;103:580). In this study, floxed-Nkx2-5 alleles were deleted using tamoxifen inducible Cre transgene (Cre-ER) beginning at 2 weeks of age. The loss of Nkx2-5 beginning at 2 weeks of age resulted in conduction and contraction defects similar to the perinatal loss of Nkx2-5, however, with a substantially slower disease progression shown by 1 degrees atrioventricular block at 6 weeks of age (4 weeks after tamoxifen injections) and heart enlargement after 12 weeks of age (10 weeks after tamoxifen injections). The phenotypes were accompanied by a slower and smaller degree of reduction of several critical Nkx2-5 downstream targets that were observed in mice with a perinatal loss of Nkx2-5. These results suggest that Nkx2-5 is necessary for proper conduction and contraction after 2 weeks of age, but with a substantially distinct level of necessity at 2 weeks of age compared with that in the perinatal period. PMID- 19546855 TI - An offer you can't refuse? Ethical implications of non-invasive prenatal diagnosis. PMID- 19546854 TI - Osteopetrosis with micro-lacunar resorption because of defective integrin organization. AB - In vitro differentiated monocytes were used to characterize the cellular defect in a type of osteopetrosis with minimally functional osteoclasts, in which defects associated with common causes of osteopetrosis were excluded by gene sequencing. Monocytes from the blood of a 28-year-old patient were differentiated in media with RANKL and CSF-1. Cell fusion, acid compartments within cells, and tartrate resistant acid phosphatase (TRAP) activity were normal. However, the osteoclasts made abnormally small pits on the dentine. Phalloidin labeling showed that the cell attachments lacked the peripheral ring structure that supports lacunar resorption. Instead, the osteoclasts had clusters of podosomes near the center of cell attachments. Antibody to the alphavbeta3 integrin pair or to the C terminal of beta3 did not label podosomes, but antibody to alphav labeled them. Western blots using antibody to the N-terminal of beta3 showed a protein of reduced size. Integrins beta1 and beta5 were upregulated, but, in contrast to observations in beta3 defects, alpha2 had not increased. The rho-GTP exchange protein Vav3, a key attachment organizing protein, did not localize normally with peripheral attachment structures. Vav3 forms of 70 kD and 90 kD were identified on western blots. However, the proteins beta3 integrin, Vav3, Plekhm1, and Src, implicated in attachment defects, had normal exon sequences. In this new type of osteopetrosis, the integrin-organizing complex is dysfunctional, and at least two attachment proteins may be partially degraded. PMID- 19546856 TI - Fitness and its role in evolutionary genetics. AB - Although the operation of natural selection requires that genotypes differ in fitness, some geneticists may find it easier to understand natural selection than fitness. Partly this reflects the fact that the word 'fitness' has been used to mean subtly different things. In this Review I distinguish among these meanings (for example, individual fitness, absolute fitness and relative fitness) and explain how evolutionary geneticists use fitness to predict changes in the genetic composition of populations through time. I also review the empirical study of fitness, emphasizing approaches that take advantage of recent genetic and genomic data, and I highlight important unresolved problems in understanding fitness. PMID- 19546858 TI - Boveri revisited: chromosomal instability, aneuploidy and tumorigenesis. AB - The mitotic checkpoint is a major cell cycle control mechanism that guards against chromosome missegregation and the subsequent production of aneuploid daughter cells. Most cancer cells are aneuploid and frequently missegregate chromosomes during mitosis. Indeed, aneuploidy is a common characteristic of tumours, and, for over 100 years, it has been proposed to drive tumour progression. However, recent evidence has revealed that although aneuploidy can increase the potential for cellular transformation, it also acts to antagonize tumorigenesis in certain genetic contexts. A clearer understanding of the tumour suppressive function of aneuploidy might reveal new avenues for anticancer therapy. PMID- 19546857 TI - Collective cell migration in morphogenesis, regeneration and cancer. AB - The collective migration of cells as a cohesive group is a hallmark of the tissue remodelling events that underlie embryonic morphogenesis, wound repair and cancer invasion. In such migration, cells move as sheets, strands, clusters or ducts rather than individually, and use similar actin- and myosin-mediated protrusions and guidance by extrinsic chemotactic and mechanical cues as used by single migratory cells. However, cadherin-based junctions between cells additionally maintain 'supracellular' properties, such as collective polarization, force generation, decision making and, eventually, complex tissue organization. Comparing different types of collective migration at the molecular and cellular level reveals a common mechanistic theme between developmental and cancer research. PMID- 19546859 TI - Rare structural variants found in attention-deficit hyperactivity disorder are preferentially associated with neurodevelopmental genes. AB - Attention-deficit/hyperactivity disorder (ADHD) is a common and highly heritable disorder, but specific genetic factors underlying risk remain elusive. To assess the role of structural variation in ADHD, we identified 222 inherited copy number variations (CNVs) within 335 ADHD patients and their parents that were not detected in 2026 unrelated healthy individuals. Although no excess CNVs, either deletions or duplications, were found in the ADHD cohort relative to controls, the inherited rare CNV-associated gene set was significantly enriched for genes reported as candidates in studies of autism, schizophrenia and Tourette syndrome, including A2BP1, AUTS2, CNTNAP2 and IMMP2L. The ADHD CNV gene set was also significantly enriched for genes known to be important for psychological and neurological functions, including learning, behavior, synaptic transmission and central nervous system development. Four independent deletions were located within the protein tyrosine phosphatase gene, PTPRD, recently implicated as a candidate gene for restless legs syndrome, which frequently presents with ADHD. A deletion within the glutamate receptor gene, GRM5, was found in an affected parent and all three affected offspring whose ADHD phenotypes closely resembled those of the GRM5 null mouse. Together, these results suggest that rare inherited structural variations play an important role in ADHD development and indicate a set of putative candidate genes for further study in the etiology of ADHD. PMID- 19546861 TI - Global proteomic profiling reveals altered proteomic signature in schizophrenia serum. AB - Schizophrenia is one of the most severe psychiatric disorders affecting 1% of the world population. There is yet no empirical method to validate the diagnosis of the disease. The identification of an underlying molecular alteration could lead to an improved disease understanding and may yield an objective panel of biomarkers to aid in the diagnosis of this devastating disease. Presented is the largest reported liquid chromatography-mass spectrometry-based proteomic profiling study investigating serum samples taken from first-onset drug-naive patients compared with samples collected from healthy volunteers. The results of this large-scale study are presented along with enzyme-linked immunosorbent assay based validation data. PMID- 19546860 TI - The dysbindin-containing complex (BLOC-1) in brain: developmental regulation, interaction with SNARE proteins and role in neurite outgrowth. AB - Previous studies have implicated DTNBP1 as a schizophrenia susceptibility gene and its encoded protein, dysbindin, as a potential regulator of synaptic vesicle physiology. In this study, we found that endogenous levels of the dysbindin protein in the mouse brain are developmentally regulated, with higher levels observed during embryonic and early postnatal ages than in young adulthood. We obtained biochemical evidence indicating that the bulk of dysbindin from brain exists as a stable component of biogenesis of lysosome-related organelles complex 1 (BLOC-1), a multi-subunit protein complex involved in intracellular membrane trafficking and organelle biogenesis. Selective biochemical interaction between brain BLOC-1 and a few members of the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) superfamily of proteins that control membrane fusion, including SNAP-25 and syntaxin 13, was demonstrated. Furthermore, primary hippocampal neurons deficient in BLOC-1 displayed neurite outgrowth defects. Taken together, these observations suggest a novel role for the dysbindin containing complex, BLOC-1, in neurodevelopment, and provide a framework for considering potential effects of allelic variants in DTNBP1--or in other genes encoding BLOC-1 subunits--in the context of the developmental model of schizophrenia pathogenesis. PMID- 19546862 TI - The role of transient receptor potential channels in kidney disease. AB - The transient receptor potential (TRP) superfamily consists, in mammals, of six protein subfamilies, TRPC, TRPM, TRPV, TRPA, TRPML and TRPP. TRPs are cation channels involved in many physiological processes and in the pathogenesis of various disorders. In the kidney, TRP channels are expressed along the nephron, and a role for some of these channels in renal function has been proposed. TRPC3 is thought to facilitate calcium ion influx into the principal cells of the collecting duct in response to vasopressin. TRPM3 and TRPV4 might be osmosensors, whereas the TRPP1/TRPP2 complex could function as a mechanosensor in the cilia of renal epithelial cells. A number of kidney diseases have also been linked to dysfunctional activity of TRPs. TRPC6 dysfunction has been associated with the onset of focal segmental glomerosclerosis; TRPP2 dysfunction is linked to autosomal-dominant polycystic kidney disease, TRPM6 mutations underlie hypomagnesemia with secondary hypocalcemia, and TRPV1 dysfunction is implicated in renal hypertension. A link between TRPC1 dysfunction and diabetic nephropathy has also been suggested in an animal model. Animal studies have implicated a role for TRPV5 in idiopathic hypercalciuria and vitamin D-dependent rickets, although these observations have not been confirmed in patients. This Review focuses on the role of renal TRP channels in health and disease. PMID- 19546863 TI - MRI as a tool to monitor islet transplantation. AB - The development of new methods for noninvasive imaging is an area of biotechnology that is of great relevance for the diagnosis and characterization of diabetes mellitus. Noninvasive imaging can be used to study the dynamics of beta-cell mass and function; beta-cell death; vascularity, innervation and autoimmune attack of pancreatic islets; and the efficacy of islet transplantation to remedy beta-cell loss in patients with diabetes mellitus. In this Review, we focus on the application of MRI for monitoring islet transplantation and on the potential causes of islet graft failure, which are still poorly understood. Questions that have been addressed by MRI studies encompass graft longevity, and the effects of immune rejection, glucose toxic effects, and the transplanted islets' purity on graft fate. We also highlight novel technologies for simultaneous imaging and delivery of experimental therapies that aim to extend the lifespan and functionality of islet grafts. On the basis of this evidence, MRI represents a valuable platform for a thorough investigation of beta-cell function in the context of islet transplantation. State-of-the-art multimodality approaches, such as PET-MRI, can extend our current capabilities and help answer the critical questions that currently inhibit the prevention and cure of diabetes mellitus. PMID- 19546864 TI - Quantitative imaging biomarkers in neuro-oncology. AB - Conventional structural imaging provides limited information on tumor characterization and prognosis. Advances in neurosurgical techniques, radiotherapy planning and novel drug treatments for brain tumors have generated increasing need for reproducible, noninvasive, quantitative imaging biomarkers. This Review considers the role of physiological MRI and PET molecular imaging in understanding metabolic processes associated with tumor growth, blood flow and ultrastructure. We address the utility of various techniques in distinguishing between tumors and non-neoplastic processes, in tumor grading, in defining anatomical relationships between tumor and eloquent brain regions and in determining the biological substrates of treatment response. Much of the evidence is derived from limited case series in individual centers. Despite their 'added value', the effect of these techniques as an adjunct to structural imaging in clinical research and practice remains limited. PMID- 19546867 TI - Parent-of-origin specific linkage and association of the IGF2 gene region with birth weight and adult metabolic risk factors. AB - OBJECTIVE: The maternally imprinted insulin-like growth factor 2 (IGF2) gene is an important fetal growth factor and is also suggested to have postnatal metabolic effects. In this study, we examined whether common polymorphisms in IGF2 (6815_6819delAGGGC, 1156T>C and 820G>A (ApaI)) and a microsatellite marker in the close vicinity of IGF2 were linked to or associated with birth weight and adult metabolic risk factors. DESIGN AND PARTICIPANTS: Polymorphisms were genotyped in 199 monozygotic complete twin pairs, 109 dizygotic complete twin pairs, 15 single twins, 231 mothers and 228 fathers recruited from the East Flanders Prospective Twin Survey. Conventional and parent-of-origin specific linkage and association analyses were carried out with birth weight, adult body height and parameters quantifying obesity, insulin sensitivity and dyslipidaemia measured at adult age (mean age 25 years). RESULTS: In the parent-of-origin specific association analysis, in which only the paternally inherited allele was incorporated, the 1156T>C SNP (single nucleotide polymorphism) showed significant association with IGF-binding protein 1 (IGFBP1) levels (T and C (mean (95% CI)): 13.2 (12.1-14.3) and 16.2 (14.6-18.0) ng ml(-1), P=0.002). No linkage was observed in either the conventional or in the parent-of-origin specific linkage analysis. CONCLUSION: This study suggests that paternally inherited alleles of a common polymorphism in the IGF2 gene affect IGFBP1 levels. PMID- 19546865 TI - Immunotherapeutic strategies in kidney cancer--when TKIs are not enough. AB - FDA approval of the multitargeted, antiangiogenic tyrosine kinase inhibitors (TKIs) sunitinib and sorafenib, and the serine and threonine mammalian target of rapamycin inhibitor, temsirolimus, has revolutionized the management of metastatic clear-cell renal-cell carcinoma (CC-RCC). The inability of these targeted therapies to provide durable complete responses, however, is a serious limiting factor to their clinical usefulness. Although immunotherapeutic approaches in advanced disease are increasingly regarded as a historical treatment paradigm, we propose that a fundamental understanding of immunobiology in CC-RCC can improve the selection of patients for high-dose intravenous interleukin 2 and facilitate the development of novel immunotherapeutic strategies. In our opinion, immunotherapeutic strategies have an important place in the management of advanced CC-RCC in the era of biological targeted therapy. PMID- 19546866 TI - Biomarkers of abdominal aortic aneurysm progression. Part 2: inflammation. AB - Defining progression of abdominal aortic aneurysm (AAA) is complicated by several factors, including measurement error, duration of follow-up, and the imaging modality used to assess AAA expansion. Investigations of biomarkers of AAA progression should be standardized so that valid comparisons can be made. Previous research has shown some promising advances towards identifying a reliable and individual predictor of AAA progression. In this second part of our Review on biomarkers of AAA progression, we examine direct and indirect markers of inflammation including various cytokines, C-reactive protein, activators of tissue plasminogen activator and urokinase plasminogen activator, and osteopontin. PMID- 19546868 TI - Physical activity patterns and prevention of weight gain in premenopausal women. AB - BACKGROUND: Studies of the association between physical activity (PA) and weight maintenance have been inconsistent. METHODS: We prospectively examined the association between PA patterns and prevention of weight gain among 46 754 healthy premenopausal women, aged 25-43 years in 1989. Participants reported their PA and weight in 1989 and 1997. The primary outcome was gaining >5% of baseline weight by 1997 (62% of the population). RESULTS: Compared with women who maintained <30 min d(-1) of total discretionary activity over 8 years, women were less likely to gain weight if they sustained 30+ min d(-1) (odds ratio (OR)=0.68, 95% confidence interval (CI) 0.64-0.73) or increased to 30+ min d(-1) in 1997 (OR=0.64, 95%CI=0.60-0.68). Among women whose only reported activity was walking, risk of gaining weight was lower in those who sustained 30+ min d(-1) over 8 years (OR=0.66, 95%CI=0.49-0.91), and brisk walking pace independently predicted less weight gain. For a 30 min d(-1) increase between 1989 and 1997, jogging/running was associated with less weight gain than brisk walking or other activities. Greater duration of PA was associated with progressively less weight gain, but even an 11-20 min d(-1) increase was beneficial; the benefits appeared stronger among those who were initially overweight. Sedentary behavior independently predicted weight gain. CONCLUSIONS: Sustained PA for at least 30 min d(-1), particularly if more intense, is associated with a reduction in long term weight gain, and greater duration is associated with less weight gain. Sedentary women of any baseline weight who increase their PA will benefit, but overweight women appear to benefit the most. PMID- 19546869 TI - The interactive effect of hunger and impulsivity on food intake and purchase in a virtual supermarket. AB - OBJECTIVE: It has been shown repeatedly that impulsivity, obesity and food intake are related; obese people are more impulsive than lean people and impulsive people eat more than less impulsive people. The relation between impulsivity and food intake might be state dependent; hunger motivates food seeking behaviour and food consumption, especially of high caloric food. Difficulties to overrule automatic behavioural tendencies might make impulsive people more susceptible to the effects of hunger on food selection. Therefore, they are expected to increase their intake more than low impulsive people when feeling hungry. STUDY 1: Fifty seven female participants were randomly assigned to a hunger or sated condition. Response inhibition (a measure of impulsivity) and food intake were measured. Results show that impulsive participants ate significantly more, but only when feeling hungry. STUDY 2: Ninety-four undergraduate students participated. Hunger, response inhibition and the purchase of food in a virtual supermarket were measured. The same interaction was found: impulsive participants bought most calories, especially from snack food, but only when feeling hungry. CONCLUSION: Hunger and impulsivity interact in their influence on consumption. These data suggest that reducing hunger during calorie restricting diets is important for successful weight loss, particularly for the impulsive dieters. PMID- 19546870 TI - Intestinal electric stimulation accelerates whole gut transit and promotes fat excrement in conscious rats. AB - INTRODUCTION: Intestinal electric stimulation (IES) is proposed as a potential tool for the treatment of morbid obesity. Our earlier study showed that IES with one pair of electrodes accelerated intestinal transit and decreased fat absorption in a segment of the jejunum in anesthetized rats. The aims of this study were to assess the effects of IES on whole gut transit and fat absorption in conscious rats, to examine the effects of multi-pairs IES and to explore the cholinergic mechanism behind the effects of IES. METHODS: Thirty-eight male rats implanted with serosal electrodes were randomized into five groups: control without IES, two- or three-pairs IES with short pulses, atropine and atropine plus IES. The whole gut transit and fat remained and emptied from the gut were analyzed after continuous 2-h IES. RESULTS: Two- and three-pairs IES significantly accelerated phenol red (PR, marker used for transit) excretion (analysis of variance (ANOVA), P<0.001). No significant difference was found between two- and three-pairs IES. Two-pairs IES significantly increased the excretion of fat (P<0.05). Atropine significantly blocked the accelerated transit induced by IES (ANOVA, P<0.001). Correlation was found between the percentage of PR and fat retained in the whole gut (r=0.497, P<0.01). CONCLUSIONS: IES accelerates whole gut transit and promotes fat excrement in conscious rats, and these effects are mediated through the cholinergic nerves. These findings are in support of the concept that IES may be a promising treatment option for obesity. PMID- 19546871 TI - Assessment of the sympathetic level of lesion in patients with spinal cord injury. PMID- 19546872 TI - Patient and caregiver knowledge of severity of injury among youth with spinal cord injury. AB - DESIGN: Multicenter cross sectional study. OBJECTIVES: Describe patient and caregiver knowledge of severity of injury and examine the relationship between AIS status and patient/caregiver report. SETTING: United States METHODS: Participants were between 1 and 21 years of age with a stable spinal cord injury (SCI). Participants underwent ISCSCI exams and were interviewed with the following questions: 1. Did the patient/caregiver know the difference between complete and incomplete spinal cord injury before participation? 2. What level of injury does the patient/caregiver report? 3. What severity of injury does the patient/caregiver report? 4. If a severity is given, who told them and how was it tested? RESULTS: Overall, 16% of patients and 20% of caregivers knew the difference between complete and incomplete SCI. Older patients were more likely to know the difference and caregivers of patients with shorter durations of injury were more likely to know the difference. Those who reported a severity of injury different from their actual severity were more likely to have a complete spinal cord injury and a higher injury severity as measured by the AIS impairment scale. Only 18% of people who were able to report a severity of injury stated that an ISCSCI exam was how the doctor gave them the diagnosis. CONCLUSION: Many patients and caregivers do not understand the difference between complete and incomplete SCI. It is vital that an AIS diagnosis only be given following the ISCSCI exam based on agreed standards. PMID- 19546873 TI - Characteristics of adults with incident traumatic spinal cord injury in Ontario, Canada. AB - STUDY DESIGN: Cohort study. OBJECTIVES: To provide recent estimates of the incidence of traumatic spinal cord injury (SCI) in adults living in Ontario. SETTING: Ontario, Canada. METHODS: The study included all men and women aged 18 years and older living in Ontario. The two primary data sources used for this study were the census data provided by Statistics Canada and the hospital Discharge Abstract Database (DAD) provided by the Canadian Institute for Health Information. Incidence was estimated for the fiscal years 2003/04-2006/07, and examined by age, gender, mechanism and seasonality of injury, the level of injury, the presence of comorbidity and in-hospital mortality. RESULTS: The incident cases had a mean age of 51.3 years (s.d. 20.1). The majority of the cases was male (74.1%) and had a cervical SCI caused by falls (49.5%). The age adjusted incidence rate was stable over the 4-year study period, from 24.2 per million (95% CI: 21.2-27.6) in 2003 to 23.1 per million (95% CI: 20.2-26.3) in 2006. CONCLUSION: Despite worldwide trends that have indicated motor vehicle collisions (MVCs) as the leading cause of injury, falls emerged as the leading cause of traumatic SCI in this study. This finding, and the fact that the number of fall-induced injuries increased steadily with age, may indicate that there is growing concern for the consequences of falls in the elderly. Further work is needed to understand this trend in age and gender and the causes of falls to develop effective fall prevention strategies. PMID- 19546874 TI - Intensity of pain and biochemical changes in blood plasma in spinal cord trauma. AB - STUDY DESIGN: Cross-sectional, observational and longitudinal. OBJECTIVES: The aim of the study was to analyze the relationship between pain intensity, plasma lipids and severity of spinal cord injuries in patients with paraplegia (n = 11), tetraplegia (n = 16) and polytrauma (n = 15). We concentrated on the hospitalization period immediately following injury. METHODS: Pain intensity was assessed on a visual analog scale immediately after patients were transported to hospital, again 14 days after injury and before discharge from hospital. Blood samples were also obtained at these same times. We measured following biochemical parameters: total protein, albumin, total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides, glycemia, and C-reactive protein. Data were analyzed with respect to type of injury, state of unconsciousness immediately after injury, hemorrhage, measure of liability (self-inflicted injuries vs casualties), cause of the accident and pre-injury cholesterol levels. RESULTS: On the day of injury, pain intensity correlated positively with HDL cholesterol (r = 0.48, P = 0.04); on the day of discharge from hospital, pain intensity correlated positively with blood glucose levels (r = 0.67, P = 0.0002). Diagnostic subgroups did not differ either in pain intensity or in pain dynamics during hospitalization. Total cholesterol level was lowest in patients with polytrauma. In all patients, the lowest total cholesterol level was observed immediately after injury. HDL cholesterol was highest after injury. CONCLUSION: After spinal cord injury, lower total cholesterol levels reflected more serious trauma intensity and HDL cholesterol predicted more intensive pain. Subjects responsible for their own injuries suffered less intensive pain than those who were not responsible for their injuries. PMID- 19546875 TI - Rhythms of serum melatonin in rats with acute spinal cord injury at the cervical and thoracic regions. AB - OBJECTIVE: To evaluate the serum melatonin levels in acute period of the spinal cord injury (SCI) caused by trauma in the rats. BACKGROUND: Traumatic SCI induces many types of physiological and pathological damage, including hormonal level variations. METHODS: Forty male Sprague-Dawley rats were divided randomly into four groups. In the control group, neither laminectomy nor SCI were performed; only a large laminectomy was performed without SCI in the sham group. In the cervical and thoracic spinal trauma groups, laminectomies at C5-6 and a T6-7 were performed, respectively, followed by clip compression of the spinal cord. Blood samples were drawn 2, 6, 12 and 24 h after the procedures and assayed immediately. RESULTS: The levels of melatonin in the neurotrauma groups were high in the first 2 h, but at the end of sixth hour, melatonin levels increased in sham-operated group, decreased in neurotrauma groups and did not change in control group. At the 12th hour, melatonin levels continued to decrease in thoracic group significantly. At the 24th hour, the melatonin levels decreased in the cervical, control and sham groups, whereas melatonin levels increased in the thoracic group. CONCLUSIONS: The present study revealed that (1) activation of endogen melatonin secretion of the organism starts immediately after the SCI, but it shows a great lowering trend between 2 and 6 h post-SCI, (2) the tetraplegic rats, which had complete injuries at the lower cervical spinal cord, could not produce enough melatonin secretion; on the contrary, the paraplegic rats, which had complete injury at the upper thoracic spinal cord, showed normal melatonin secretion. PMID- 19546876 TI - Effect of wearing an activity monitor on the amount of daily manual wheelchair propulsion in persons with spinal cord injury. AB - STUDY DESIGN: An experimental study. OBJECTIVES: To assess the effect of reactivity related to wearing a multi-sensor activity monitor (AM) on the amount of manual wheelchair propulsion during daily life in wheelchair-bound persons with spinal cord injury (SCI). In addition, to establish the subjectively experienced burden of wearing the AM. SETTING: Rehabilitation centre and home based study. METHODS: In 10 persons with SCI, during a 7-day period, the daily amount of manual wheelchair propulsion was measured by means of a rotation counter. During this period, an AM was worn for 1 day (AM+ day) by the participants. Experienced burden was measured by a questionnaire based on visual analogue scale scores. RESULTS: The overall median of the number of rotations per minute was 1.38 (range 0.63-1.83). No significant difference was found in the amount of daily manual wheelchair propulsion between AM+ and AM- days (P=0.33, median difference: -0.06 rotations per minute). Experienced burden was not different between subgroups that differed in reactivity. CONCLUSION: The results seem to indicate that wearing the AM of this study does not systematically influence the amount of daily manual wheelchair propulsion. Although low to moderate burden was experienced when wearing the AM, this does not seem to affect the amount of manual wheelchair propulsion. PMID- 19546877 TI - Health care utilization in non-traumatic and traumatic spinal cord injury: a population-based study. AB - STUDY DESIGN: Retrospective cohort design. OBJECTIVES: To compare socio demographic, impairment characteristics and utilization (physician and emergency department (ED) visits) for non-traumatic (NTSCI) and traumatic (TSCI) spinal cord injury 1 year post inpatient rehabilitation. SETTING: Ontario, Canada. METHODS: Inpatient stays (2003-2006) were identified from the National Rehabilitation Registry System. Exclusions were: in-hospital mortality; discharge after 31 March 2006; death within 1 year after discharge. Multivariate logistic regression analyses were used to determine factors predicting high utilization. RESULTS: NTSCI cases (n=1002) were greater than TSCI (n=560). NTSCIs were older (mean=61.6, s.d.=15.8) with more co-morbidities, paraplegic (39.5%) and female (P<0.001). NTSCI had higher FIM admission and discharge scores but lower change scores. Mean number of physician visits for NTSCI and TSCI were 31.2 (median=24) and 29.7 (median=22), with no significant differences in mean specialist visits (NTSCI 16.5: TSCI 17.0). Factors predicting 30 or more physician visits included age 60 years or above (OR=1.5; 95% CI=1.2-1.9), urban living (OR=1.59; 95% CI=1.12-2.22) and lowest quartile (18-88) discharge FIM (OR=1.8; 95% CI=1.4-2.3). Charlson score of 3 or more (OR=2.1; 95% CI=1.3-3.2), urban living (OR=1.92; 95% CI=1.3-2.86) and lowest quartile discharge FIM (OR=1.5; 95% CI=1.2-2.0) were associated with 20 or more specialist visits. Factors for high ED use were: rurality (OR=1.5; 95% CI=1.1-2.1), low income (OR=1.4; 95% CI=1.1-1.9) and low (18-88) discharge FIM (OR=1.7; 95% CI=1.3-2.2). CONCLUSION: Both demonstrated significant health care utilization requiring attention to health care needs; particularly for those living in rural settings, with low income and/or low functional ability. PMID- 19546878 TI - NK cells enhance the induction of CTL responses by IL-15 monocyte-derived dendritic cells. AB - Dendritic cells differentiated from monocytes (MoDC) in the presence of GM-CSF and IL-15 (IL-15 MoDC) exhibit superior migration and cytotoxic T-lymphocyte (CTL) induction compared with MoDC differentiated in IL-4 and GM-CSF (IL-4 MoDC) and are promising candidates for DC immunotherapy. We explored the mechanisms by which IL-15 MoDC induce CTL. IL-15 MoDC expressed higher levels of CD40 and secreted high levels of TNF-alpha, but little or no IL-12p70 compared with IL-4 MoDC. Despite immuno-selecting monocytes to >97% purity before MoDC generation, a tiny population (0.2%) of natural killer (NK) cells was identified that was increased sevenfold during IL-15 MoDC, but not IL-4 MoDC differentiation. These NK cells produced high levels of IFN-gamma and were responsible for the enhanced CTL-inducing capacity of the IL-15 MoDC, but not for their increased expression of CD40 or secretion of TNF-alpha. Interestingly, a proportion of IL-15 MoDC were found to express the NK cell marker, CD56, but these did not secrete IFN-gamma. These data implicate a role for small percentages of NK cells in the enhanced capacity of IL-15 MoDC to induce tumour-specific CTL independent of IL-12p70. PMID- 19546880 TI - Influence of ethnicity on pharmacogenetic variation in the Ghanaian population. AB - It has been well established that the frequencies of genomic variants can vary greatly between the populations of different countries. We sought to quantify the intra-population variability in Ghana to determine the value of genotyping studies done at a nationwide level. Further, we investigated the differences between the Ghanaian and other African populations to determine the quality of genomic representation provided by a small subgroup within the continent with regard to the general population. We genotyped 934 unrelated Ghanaian individuals for 15 single nucleotide polymorphisms (SNPs) from genes defined as clinically relevant based on their reported roles in the transport of, metabolism of, or as targets of the medicines listed in the World Health Organization Essential Medicines list. Populations within Ghana and between nations in Western Africa were genetically cohesive. In contrast, populations in other areas of Africa were genetically divergent. Gene allele frequency also differed significantly between the populations in African nations and the United States for several of the SNPs. These results demonstrate that national populations in similar geographic regions, like Africa, may have widely varying genetic allele frequencies for clinically relevant SNPs. Further genotyping studies of specific populations are necessary to provide the best medical care to all individuals. PMID- 19546881 TI - Alterations of gene expression in blood cells associated with chronic fatigue in breast cancer survivors. AB - Fatigue is one of the most frequent complaints among breast cancer survivors. However, mechanisms underlying persisting fatigue after end of treatment are poorly understood. To explore whether biological processes underlying persistent fatigue can affect gene expression of blood cells, genome-wide expression analyses were performed on whole blood samples from breast cancer survivors classified as chronic fatigued 2-6 years after diagnosis. Non-fatigued survivors served as controls. Several gene sets involved in plasma- and B-cell pathways differed between the chronic fatigued and the non-fatigued, suggesting that a dysregulation in these pathways is associated with chronic fatigue and that a B cell-mediated inflammatory process might underlie fatigue. The chronic fatigued also had a higher level of leucocytes, lymphocytes and neutrophiles compared with the non-fatigued, thus further indicating that an activation of the immune system plays a role in the biology of chronic fatigue in breast cancer survivors. PMID- 19546879 TI - Why is effective treatment of asthma so difficult? An integrated systems biology hypothesis of asthma. AB - A hypothesis is presented that asthma is not only an airway disease, but that the disease involves the entire lung, and that the chronicity of asthma and asthma exacerbations can perhaps be explained if one considers asthma as a systemic disease. Increased lung-not only airway-vascularity may be the result of the action of angiogenesis factors, such as vascular endothelial growth factor (VEGF) and sphingosine-1-phosphate (S1P). A bone-marrow lung axis can be postulated as one element of the systemic nature of the asthma syndrome, in which the inflamed lung emits chemotactic signals, which the bone marrow responds to by releasing cells that contribute to lung angiogenesis. A molecular model of the pathobiology of asthma can be built by connecting hypoxia-inducible transcription factor-1 alpha, VEGF S1P, and bone-marrow precursor cell mobilization and acknowledging that angiogenesis is part of the inflammatory response. PMID- 19546882 TI - Lymph node dissection during robot-assisted radical prostatectomy: where do we stand? AB - Since the initial report of robot-assisted laparoscopic prostatectomy (RALP) in 2001, the technique has gained rapid acceptance and utilization. When compared with more traditional forms of surgical intervention, there is still much debate with respect to cost, and impact on potency and continence. Less often is the focus on oncologic outcomes. Pelvic lymph node dissection (PLND) at the time of prostatectomy is an important part of the surgical intervention for prostate cancer and is currently underreported during robotic procedures. Herein, we review the current controversies on the value and extent of PLND and the status of emerging data regarding robot-assisted PLND. PMID- 19546883 TI - Androgens induce oxidative stress and radiation resistance in prostate cancer cells though NADPH oxidase. AB - Androgen deprivation therapy (ADT) facilitates the response of prostate cancer (PC) to radiation. Androgens have been shown to induce elevated basal levels of reactive oxygen species (ROS) in PC, leading to adaptation to radiation-induced cytotoxic oxidative stress. Here, we show that androgens increase the expression of p22(phox) and gp91(phox) subunits of NADPH oxidase (NOX) and ROS production by NOX2 and NOX4 in PC. Pre-radiation treatment of 22Rv1 human PC cells with NOX inhibitors sensitize the cells to radiation similarly to ADT, suggesting that their future usage may spare the need for adjuvant ADT in PC patients undergoing radiation. PMID- 19546884 TI - Subjective and objective measures of late genitourinary morbidity following hypofractionated radiotherapy in men with prostate cancer. AB - To value the late genitourinary (GU) morbidity in men treated with a hypofractionated radiotherapy regimen for prostate cancer. Patients with intermediate risk factors according to D'Amico's criteria were selected. The hypofractionated schedule consisted of 15 fractions of 3.63 Gy delivered three times per week for a total dose of 54.3 Gy. Significant changes in storage symptoms were not found. A significant transient worsening in the score of late effects of normal tissue late effects normal tissue task force (LENT)-subjective, objective, management, analytic (SOMA) urinary-function domain was observed at 12 months with subsequent improvement at 28 months. The assessment of voiding symptoms and maximum urinary flow rate (Qmax) showed that no significant difference was measurable at 12 and 28 months. For PVR, a transient increase at 12 months with a subsequent decrease at 28 months was measured. No significant increase in alpha-blockers usage and in the percentage of men with pathological nonintubated uroflowmetry (NIF) was observed at 12 and 28 months. Finally, patients did not perceive any clinical worsening in their quality of life (QoL) as attested by the International Prostate Symptom Score (IPSS)-QoL. Our study seems to suggest that our hypofractionated radiotherapy schedule for the treatment of prostate cancer is safe in terms of late urinary morbidity. Further study will be required to confirm our results. PMID- 19546885 TI - Berberine promotes the development of atherosclerosis and foam cell formation by inducing scavenger receptor A expression in macrophage. AB - Berberine is identified to lower the serum cholesterol level in human and hamster through the induction of low density lipoproteins (LDL) receptor in hepatic cells. To evaluate its potential in preventing atherosclerosis, the effect of berberine on atherosclerosis development in apolipoprotein E-deficient (apoE(-/ )) mice was investigated. In apoE(-/-) mice, berberine induced in vivo foam cell formation and promoted atherosclerosis development. The foam cell formation induced by berberine was also observed in mouse RAW264.7 cells, as well as in mouse and human primary macrophages. By inducing scavenger receptor A (SR-A) expression in macrophages, berberine increased the uptake of modified LDL (DiO-Ac LDL). Berberine-induced SR-A expression was also observed in macrophage foam cells in vivo and in the cells at atherosclerotic lesion. Analysis in RAW264.7 cells indicated that berberine induced SR-A expression by suppressing PTEN expression, which led to sustained Akt activation. Our results suggest that to evaluate the potential of a cholesterol-reducing compound in alleviating atherosclerosis, its effect on the cells involved in atherosclerosis development, such as macrophages, should also be considered. Promotion of foam cell formation could counter-balance the beneficial effect of lowering serum cholesterol. PMID- 19546886 TI - Regulation of the cell cycle gene, BTG2, by miR-21 in human laryngeal carcinoma. AB - MicroRNAs are short regulatory RNAs that negatively modulate gene expression at the post-transcriptional level, and are deeply involved in the pathogenesis of several types of cancers. To investigate whether specific miRNAs and their target genes participate in the molecular pathogenesis of laryngeal carcinoma, oligonucleotide microarrays were used to assess the differential expression profiles of microRNAs and mRNAs in laryngeal carcinoma tissues compared with normal tissues. The oncogenic miRNA, microRNA-21 (miR-21), was found to be upregulated in laryngeal carcinoma tissues. Knockdown of miR-21 by specific antisense oligonucleotides inhibited the proliferation potential of HEp-2 cells, whereas overexpression of miR-21 elevated growth activity of the cells, as detected by the colony formation assay. The cell number reduction caused by miR 21 inhibition was due to the loss of control of the G1-S phase transition, instead of a noticeable increase in apoptosis. Subsequently, a new target gene of miR-21, BTG2, was found to be downregulated in laryngeal carcinoma tissues. BTG2 is known to act as a pan-cell cycle regulator and tumor suppressor. These findings indicate that aberrant expression of miR-21 may contribute to the malignant phenotype of laryngeal carcinoma by maintaining a low level of BTG2. The identification of the oncogenic miR-21 and its target gene, BTG2, in laryngeal carcinoma is potentially valuable for cancer diagnosis and therapy. PMID- 19546887 TI - A design of multi-source samples as a shared control for association studies in genetically stratified populations. PMID- 19546889 TI - Lipid accumulation in hepatocytes induces fibrogenic activation of hepatic stellate cells. AB - Despite the initial belief that non-alcoholic fatty liver disease is a benign disorder, it is now recognized that fibrosis progression occurs in a significant number of patients. Furthermore, hepatic steatosis has been identified as a risk factor for the progression of hepatic fibrosis in a wide range of other liver diseases. Here, we established an in vitro model to study the effect of hepatic lipid accumulation on hepatic stellate cells (HSCs), the central mediators of liver fibrogenesis. Primary human hepatocytes were incubated with the saturated fatty acid palmitate to induce intracellular lipid accumulation. Subsequently, human HSCs were incubated with conditioned media (CM) from steatotic or control hepatocytes. Lipid accumulation in hepatocytes induced the release of factors that accelerated the activation and proliferation of HSC, and enhanced their resistance to apoptosis, largely mediated via activation of the PI-3-kinase pathway. Furthermore, CM from steatotic hepatocytes induced the expression of the profibrogenic genes TGF-beta, tissue inhibitor of metallo-proteinase-1 (TIMP-1), TIMP-2 and matrix-metallo-proteinase-2, as well as nuclear-factor kappaB dependent MCP-1 expression in HSC. In summary, our in vitro data indicate a potential mechanism for the pathophysiological link between hepatic steatosis and fibrogenesis in vivo. Herewith, this study provides an attractive in vitro model to study the molecular mechanisms of steatosis-induced fibrogenesis, and to identify and test novel targets for antifibrotic therapies in fatty liver disease. PMID- 19546888 TI - Cbl negatively regulates JNK activation and cell death. AB - Here, we explore the role of Cbl proteins in regulation of neuronal apoptosis. In two paradigms of neuron apoptosis - nerve growth factor (NGF) deprivation and DNA damage - cellular levels of c-Cbl and Cbl-b fell well before the onset of cell death. NGF deprivation also induced rapid loss of tyrosine phosphorylation (and most likely, activation) of c-Cbl. Targeting c-Cbl and Cbl-b with siRNAs to mimic their loss/inactivation sensitized neuronal cells to death promoted by NGF deprivation or DNA damage. One potential mechanism by which Cbl proteins might affect neuronal death is by regulation of apoptotic c-Jun N-terminal kinase (JNK) signaling. We demonstrate that Cbl proteins interact with the JNK pathway components mixed lineage kinase (MLK) 3 and POSH and that knockdown of Cbl proteins is sufficient to increase JNK pathway activity. Furthermore, expression of c-Cbl blocks the ability of MLKs to signal to downstream components of the kinase cascade leading to JNK activation and protects neuronal cells from death induced by MLKs, but not from downstream JNK activators. On the basis of these findings, we propose that Cbls suppress cell death in healthy neurons at least in part by inhibiting the ability of MLKs to activate JNK signaling. Apoptotic stimuli lead to loss of Cbl protein/activity, thereby removing a critical brake on JNK activation and on cell death. PMID- 19546890 TI - discs large regulates somatic cyst cell survival and expansion in Drosophila testis. AB - Gonad development requires a coordinated soma-germline interaction that ensures renewal and differentiation of germline and somatic stem cells to ultimately produce mature gametes. The Drosophila tumour suppressor gene discs large (dlg) encodes a septate junction protein functioning during epithelial polarization, asymmetric neuroblast division, and formation of neuromuscular junctions. Here, we report the role of dlg in testis development and its critical function in somatic cyst cells (SCCs). In these cells dlg is primarily required for their survival and expansion, and contributes to spermatocyte cyst differentiation. Cell death primarily occurred in SCCs at the end of spermatogonial amplification at a time when Dlg becomes restricted in wild-type (wt) testes to the distal somatic cells capping the growing spermatocyte cysts. RNAi depletion of dlg transcripts in early SCCs fully prevented testis development, whereas depletion in late SCCs resulted in a breakdown of spermatocyte cyst structure and germ cell individualization. Specific dlg expression in SCCs resulted in developmental rescue of dlg mutant testes, whereas its expression in germ cells exerted no such effect. dlg overexpression in wt testes led to spermatocyte cyst expansion at the expense of spermatogonial cysts. Our data demonstrate that dlg is essentially required in SCCs for their survival, expansion, and differentiation, and for the encapsulation of the germline cells. PMID- 19546892 TI - All-optical transversal filter with tap doubling and negative coefficients based on polarization modulation. AB - A novel all-optical transversal filter with negative coefficients and tap doubling is proposed and experimentally demonstrated. Based on polarization modulation using the anisotropy of the electro-optic coefficient of LiNbO(3) crystals, negative and positive coefficients are simultaneously generated. Tap doubling is also achieved by using wavelength- and polarization-dependent time delays in fiber. Due to the orthogonal polarity between the two coefficients of each wavelength, the proposed filter is free from coherent interference and the synthesis of filter taps is performed in the optical domain. The experimental results demonstrated that stable 6-tap bandpass filter characteristics can be obtained by using only three optical sources. PMID- 19546891 TI - Adventitious root formation in rice requires OsGNOM1 and is mediated by the OsPINs family. AB - The fibrous root system in cereals comprises primarily adventitious roots (ARs), which play important roles in nutrient and water uptake. Current knowledge regarding the molecular mechanism underlying AR development is still limited. We report here the isolation of four rice (Oryza sativa L.) mutants, from different genetic backgrounds, all of which were defective in AR formation. These mutants exhibited reduced numbers of lateral roots (LRs) and partial loss of gravitropism. The mutants also displayed enhanced sensitivity to N-1 naphthylphthalamic acid, an inhibitor of polar auxin transport (PAT), indicating that the mutations affected auxin transport. Positional cloning using one of the four mutants revealed that it was caused by loss-of-function of a guanine nucleotide exchange factor for ADP-ribosylation factor (OsGNOM1). RT-PCR and analysis of promoter::GUS transgenic plants showed that OsGNOM1 is expressed in AR primordia, vascular tissues, LRs, root tips, leaves, anthers and lemma veins, with a distribution pattern similar to that of auxin. In addition, the expressions of OsPIN2, OsPIN5b and OsPIN9 were altered in the mutants. Taken together, these findings indicate that OsGNOM1 affects the formation of ARs through regulating PAT. PMID- 19546893 TI - An improved Monte Carlo diffusion hybrid model for light reflectance by turbid media. AB - This paper introduces an improved diffusion model which is accurate and fast in computation for the cases of mu(a)/mu'(s) < 0.07 as good as the conventional diffusion model for the cases of mu(a)/mu'(s) < 0.007 for surface measurement, hence more suitable than the conventional model to be the forward model used in the image reconstruction in the diffuse optical tomography. Deviation of the diffusion approximation (DA) on the medium surface is first studied in the Monte Carlo (MC) diffusion hybrid model for reflectance setup. A modification of DA and an improved MC diffusion hybrid model based on this modified DA are introduced. Numerical tests show that for media with relatively strong absorption the present modified diffusion approach can reduce the surface deviation significantly in both the hybrid and pure diffusion model, and consumes nearly no more computation time than the conventional diffusion approach. PMID- 19546894 TI - Intracavity pulse dynamics and stability for passively mode-locked lasers. AB - We derive a general characterization of the intracavity pulse dynamics for passively mode-locked fiber lasers based on the use of the variational principle. As a first application this method is used for an efficient simulation of the laser dynamics of stretched pulse and similariton lasers and evaluation of its stability. PMID- 19546895 TI - Tunable dual-wavelength erbium-doped fiber laser stabilized by four-wave mixing in a 35-cm highly nonlinear bismuth-oxide fiber. AB - Using four-wave mixing in a 35-cm highly nonlinear bismuth-oxide fiber incorporated in an erbium-doped fiber laser, a stable dual-wavelength output is obtained. The spectral spacing has been tuned from 1.3 to 7.2 nm with a tunable fiber Bragg grating. Simultaneous tuning of the two wavelengths over 20 nm is also demonstrated using a tunable bandpass filter together with a birefringent filter defining a 100-GHz frequency comb. The output stability has been experimentally analyzed. An abrupt reduction in the intensity fluctuation is observed when the amplifier output power reaches 22.0 dBm. At 22.3 dBm, the fluctuation attains a lower limit of approximately 1 dB. PMID- 19546896 TI - Variable-focus liquid lens. AB - A variable-focus liquid lens using a transparent flat cell is demonstrated. The top substrate has an aperture which is sealed with a thin elastic membrane on the outer surface and the bottom aperture is sealed on the inner surface of the bottom substrate. By applying a pressure to the outer membrane inward causes the liquid to redistribute and swell the inner membrane outward which, in turn, forms a plano-convex lens. To prove principles, a water lens with 5 mm aperture was fabricated. The resolution is better than 25 lp/mm and the response time is approximately 40 ms. Potential applications of such a lens for real-time active imaging are emphasized. PMID- 19546897 TI - Deposition of tungsten nanogratings induced by a single femtosecond laser beam. AB - Tungsten nanogratings with sub-100nm linewidths and subwavelength periods are fabricated by laser-induced chemical vapor deposition using a single 400 nm femtosecond pulsed laser beam without any beam shaping. Combining advantages of parallel and direct-write processing, this method can produce various nanograting structures on a wide range of substrates in a single step. PMID- 19546898 TI - Design of mid-IR and THz quantum cascade laser cavities with complete TM photonic bandgap. AB - We present the design of mid-infrared and THz quantum cascade laser cavities formed from planar photonic crystals with a complete in-plane photonic bandgap. The design is based on a honeycomb lattice, and achieves a full in-plane photonic gap for transverse-magnetic polarized light while preserving a connected pattern for efficient electrical injection. Candidate defects modes for lasing are identified. This lattice is then used as a model system to demonstrate a novel effect: under certain conditions - that are typically satisfied in the THz range a complete photonic gap can be obtained by the sole patterning of the top metal contact. This possibility greatly reduces the required fabrication complexity and avoids potential damage of the semiconductor active region. PMID- 19546899 TI - Electrically-pumped, broad-area, single-mode photonic crystal lasers. AB - Planar broad-area single-mode lasers, with modal widths of the order of tens of microns, are technologically important for high-power applications and improved coupling efficiency into optical fibers. They may also find new areas of applications in on-chip integration with devices that are of similar size scales, such as for spectroscopy in microfluidic chambers or optical signal processing with micro-electromechanical systems. An outstanding challenge is that broad-area lasers often require external means of control, such as injection-locking or a frequency/spatial filter to obtain single-mode operation. In this paper, we propose and demonstrate effective index-guided, large-area, edge-emitting photonic crystal lasers driven by pulsed electrical current injection at the optical telecommunication wavelength of 1550 nm. By suitable design of the photonic crystal lattice, our lasers operate in a single mode with a 1/e(2) modal width of 25 microm and a length of 600 microm. PMID- 19546900 TI - Nonlinear silicon-on-insulator waveguides for all-optical signal processing. AB - Values up to gamma=7 x 10(6)/(W km) for the nonlinear parameter are feasible if silicon-on-insulator based strip and slot waveguides are properly designed. This is more than three orders of magnitude larger than for state-of-the-art highly nonlinear fibers, and it enables ultrafast all-optical signal processing with nonresonant compact devices. At lambda=1.55 microm we provide universal design curves for strip and slot waveguides which are covered with different linear and nonlinear materials, and we calculate the resulting maximum gamma. PMID- 19546901 TI - Two-dimensional photonic crystals constructed with a portion of photonic quasicrystals. AB - Photonic quasicrystals (PQs) can produce interesting photonic properties but the lack of periodicity in structures makes exact prediction on their photonic band structures (PBSs) a fundamental challenge currently. Here, we propose a kind of complex two-dimensional photonic crystal (PC) structures constructed with a small portion of different PQs for the purposes of overcoming the difficulty of numerical calculations on the PBSs but maintaining photonic properties of the original PQs owned. Theoretically calculated results on PBSs of the complex PC with a local feature consistent with 12-fold rotational symmetry show that, in the cases of dielectric cylinders in air, air-holes in a dielectric, and metal cylinders in air, respectively, the complex PC can indeed produce similar photonic properties of the original 12-fold PQ such as uniform or isotropic PBGs under much lower dielectric contrast etc. The complex PCs can be constructed with the local parts of n-fold symmetric PQs and should provide a way for creating novel photonic functional materials. PMID- 19546902 TI - Chromosomal analysis and identification based on optical tweezers and Raman spectroscopy: comment. AB - The authors of the work: 'Chromosomal analysis and identification based on optical tweezers and raman spectroscopy' [Opt. Express 14, 5385 (2006], claim that they have been able to identify and differentiate between three human chromosomes with an optical-tweezer - Raman Spectroscopic experimental (LTRS) set up. The results and conclusions as they are presented in the paper are questionable, however, when the spectral data and data analysis are studied in greater detail. PMID- 19546903 TI - Chromosomal analysis and identification based on optical tweezers and Raman spectroscopy: reply. AB - We appreciate the authors' comments in their reply: "On the identification of chromosomes with Raman spectroscopy: a critical comment" [Opt. Express 15, 5997 (2007)]. Their main concern with our paper is asking if the collected spectra have shown the identification or differentiation between three human chromosomes. We think this comment is flawed because the authors misunderstood the main points of the original paper and interpreted the presented spectra data (Fig. 3 and Table 1) incorrectly. PMID- 19546904 TI - System characterization of a passive 40 Gb/s All Optical Clock Recovery ahead of the receiver. AB - We report on a passive all-optical clock recovery technique based on data signal filtering with a Fabry-Perot filter, tested in a 40 Gb/s transmission system. We have simulated the clock recovery principle to choose the filter finesse and then investigate with experiment the method for 43 Gbit/s RZ signal clock recovery ahead of a receiver. We use Bit Error Rate assessment to demonstrate its system compatibility and to evaluate both its pattern sequence length tolerance and, for the first time, its clock locking range. PMID- 19546905 TI - Dispersion of nonlinear dielectric function of Au nanoparticles in silica glass. AB - We have investigated wavelength dispersion of photo-induced nonlinear dielectric function of Au nanoparticle materials. Transient transmission and reflection spectra were sequentially measured by the pump-probe method with a femtosecond laser system. The dispersion of real and imaginary parts of the nonlinear dielectric function of Au:SiO(2) nanoparticle material in the vicinity of the surface plasmon resonance was evaluated from these transient spectra with total differential. A local electromagnetic field factor and interband transition in Au nanoparticles directly dominate the dispersion. PMID- 19546906 TI - Self ordering threshold and superradiant backscattering to slow a fast gas beam in a ring cavity with counter propagating pump. AB - We study the dynamics of a fast gaseous beam in a high Q ring cavity counter propagating a strong pump laser with large detuning from any particle optical resonance. As spontaneous emission is strongly suppressed the particles can be treated as polarizable point masses forming a dynamic moving mirror. Above a threshold intensity the particles exhibit spatial periodic ordering enhancing collective coherent backscattering which decelerates the beam. Based on a linear stability analysis in their accelerated rest frame we derive analytic bounds for the intensity threshold of this selforganization as a function of particle number, average velocity, kinetic temperature, pump detuning and resonator linewidth. The analytical results agree well with time dependent simulations of the N-particle motion including field damping and spontaneous emission noise. Our results give conditions which may be easily evaluated for stopping and cooling a fast molecular beam. PMID- 19546907 TI - Characteristics of focused soft X-ray free-electron laser beam determined by ablation of organic molecular solids. AB - A linear accelerator based source of coherent radiation, FLASH (Free-electron LASer in Hamburg) provides ultra-intense femtosecond radiation pulses at wavelengths from the extreme ultraviolet (XUV; lambda<100nm) to the soft X-ray (SXR; lambda<30nm) spectral regions. 25-fs pulses of 32-nm FLASH radiation were used to determine the ablation parameters of PMMA - poly (methyl methacrylate). Under these irradiation conditions the attenuation length and ablation threshold were found to be (56.9+/-7.5) nm and approximately 2 mJ*cm(-2), respectively. For a second wavelength of 21.7 nm, the PMMA ablation was utilized to image the transverse intensity distribution within the focused beam at mum resolution by a method developed here. PMID- 19546908 TI - A hybrid AlGaInAs-silicon evanescent waveguide photodetector. AB - We report a waveguide photodetector utilizing a hybrid waveguide structure consisting of AlGaInAs quantum wells bonded to a silicon waveguide. The light in the hybrid waveguide is absorbed by the AlGaInAs quantum wells under reverse bias. The photodetector has a fiber coupled responsivity of 0.31 A/W with an internal quantum efficiency of 90 % over the 1.5 mum wavelength range. This photodetector structure can be integrated with silicon evanescent lasers for power monitors or integrated with silicon evanescent amplifiers for preamplified receivers. PMID- 19546909 TI - Electron wave-packet dynamics in a relativistic electromagnetic field: 3-D analytical approximation. AB - A solution to the Klein Gordon equation for a laser-driven electron is constructed from a superposition of Volkov states. The time-and space-dependent three-dimensional superposition integral can be evaluated analytically for an initial Gaussian momentum distribution when the expression for relativistic energy is expanded in a Taylor series over the scaled initial momenta. The solution preserves many complicated wave-packet dynamics in a strong field, including so-called wave-packet shearing and the formation of multiple peaks when the wave packet spreads to the scale of the driving-field wavelength. The range of applicability of the solution applies to much of the parameter space accessible by current intense ultra-short laser systems. PMID- 19546910 TI - Transmission area and correlated imaging. AB - The relationship between transmission area of an object imaged and the visibility of correlated imaging is investigated in a lensless system. We show that they are not in simple inverse proportion, as usually depicted. The changes of the visibility will be quite different when the transmission area is varied by different manners, which may motivate people to seek a new understanding about the influence factors of the visibility. PMID- 19546911 TI - Femtosecond laser induced space-selective precipitation of nonlinear optical crystals in rare-earth-doped glasses. AB - We report on space-selective precipitation of nonlinear optical crystals in an Er(3+)-doped BaO-TiO(2)-SiO(2) glass by using a focused femtosecond laser with 800 nm, 250 kHz and 150 fs. An intense green emission due to upconversion luminescence is observed around the focal point of the femtosecond laser beam at the initial stage of the laser irradiation. A blue emission due to second harmonic generation begins to emerge from the irradiation region after 40 s irradiation. Micro-Raman spectra indicate that nonlinear optical crystals (Ba(2)TiSi(2)O(8)) are precipitated after the laser irradiation. The irradiation time for crystallization in Er(3+)-doped BaO-TiO(2)-SiO(2) glasses is longer than that in BaO-TiO(2)-SiO(2) glasses under the same irradiation conditions. The mechanisms responsible for the observed phenomena are discussed. PMID- 19546912 TI - Miniaturized high-NA focusing-mirror multiple optical tweezers. AB - An array of high numerical aperture parabolic micromirrors (NA = 0.96) is used to generate multiple optical tweezers and to trap micron-sized dielectric particles in three dimensions within a fluidic device. The array of micromirrors allows generating arbitrarily large numbers of 3D traps, since the whole trapping area is not restricted by the field-of-view of the high-NA microscope objectives used in traditional tweezers arrangements. Trapping efficiencies of Q(max) r approximately = 0.22, comparable to those of conventional tweezers, have been measured. Moreover, individual fluorescence light from all the trapped particles can be collected simultaneously with the high-NA of the micromirrors. This is demonstrated experimentally by capturing more than 100 fluorescent micro-beads in a fluidic environment. Micromirrors may easily be integrated in microfluidic devices, offering a simple and very efficient solution for miniaturized optical traps in lab-on-a-chip devices. PMID- 19546913 TI - Comparison of surface plasmon resonance devices for acoustic wave detection in liquid. AB - Surface plasmon resonance (SPR) sensors represent a suitable method for broadband acoustic pulse detection. The reflectivity and phase of a p-polarized laser beam incident on an optical device under SPR conditions are strongly dependent on ambient conditions that are changed by an acoustic wave. Depending on the order of layers, SPR sensors can be arranged in the Kretschmann or in the Otto configuration acting as a pressure or as a displacement sensor. The aim of this study was to compare both configurations and to find linear and sensitive conditions for the application. Numerical calculations were carried out varying the layer dimensions and the angle of incidence. The results of the experimental investigation on both configurations confirm the working principle. PMID- 19546914 TI - Photoluminescence dependence of InGaN/GaN QW on embedded AlGaN delta-layer. AB - An investigation of the effects of an embedded AlGaN delta-layer on the photoluminescence (PL) efficiency of an InGaN/GaN single quantum well (SQW) is presented. In particular, we focus on the dependence of the overlap integral between the electron and hole envelope wavefunctions on the AlGaN delta-layer thickness, aluminum composition, and Mg delta-doping concentration. We have demonstrated by means of simulation and experiment that the overlap integral and PL efficiency are enhanced with increasing delta-layer thickness. They have been shown to be raised further by increasing aluminum composition and Mg delta-doping concentration in the delta-layer. PMID- 19546915 TI - Compact and efficient injection of light into band-edge slow-modes. AB - We design compact (a few wavelength long) and efficient (>99%) injectors for coupling light into slow Bloch modes of periodic thin film stacks and of periodic slab waveguides. The study includes the derivation of closed-form expressions for the injection efficiency as a function of the group-velocity of injected light, and the proof that 100% coupling efficiencies for arbitrary small group velocities is possible with an injector length scaling as log(c/vg). The trade off between the injector bandwidth and the group velocity of the injected light is also considered. PMID- 19546916 TI - Differentiated service in OBS networks using a dynamic FDL bank partitioning algorithm. AB - In order to solve the differentiated service problem in optical burst switching (OBS) networks, we propose a dynamic fiber delay line (FDL) bank partitioning algorithm, which divides a FDL bank into several groups, using a feed-forward output buffering architecture. In the analysis, three classes and groups are considered for traffic and FDL, respectively, and each group is assigned to each class. This paper shows that the loss differentiation in OBS networks is easily accomplished in Poisson traffic environments when our dynamic algorithm is adopted. PMID- 19546917 TI - In vivo high-contrast imaging of deep posterior eye by 1-microm swept source optical coherence tomography and scattering optical coherence angiography. AB - Retinal, choroidal and scleral imaging by using swept-source optical coherence tomography (SS-OCT) with a 1-microm band probe light, and high-contrast and three dimensional (3D) imaging of the choroidal vasculature are presented. This SS-OCT has a measurement speed of 28,000 A-lines/s, a depth resolution of 10.4 microm in tissue, and a sensitivity of 99.3 dB. Owing to the high penetration of the 1 microm probe light and the high sensitivity of the system, the in vivo sclera of a healthy volunteer can be observed. A software-based algorithm of scattering optical coherence angiography (S-OCA) is developed for the high-contrast and 3D imaging of the choroidal vessels. The S-OCA is used to visualize the 3D choroidal vasculature of the in vivo human macula and the optic nerve head. Comparisons of S-OCA with several other angiography techniques including Doppler OCA, Doppler OCT, fluorescein angiography, and indocyanine green angiography are also presented. PMID- 19546918 TI - Regularity based descriptor computed from local image oscillations. AB - This work presents a novel local image descriptor based on the concept of pointwise signal regularity. Local image regions are extracted using either an interest point or an interest region detector, and discriminative feature vectors are constructed by uniformly sampling the pointwise Holderian regularity around each region center. Regularity estimation is performed using local image oscillations, the most straightforward method directly derived from the definition of the Holder exponent. Furthermore, estimating the Holder exponent in this manner has proven to be superior, in most cases, when compared to wavelet based estimation as was shown in previous work. Our detector shows invariance to illumination change, JPEG compression, image rotation and scale change. Results show that the proposed descriptor is stable with respect to variations in imaging conditions, and reliable performance metrics prove it to be comparable and in some instances better than SIFT, the state-of-the-art in local descriptors. PMID- 19546919 TI - Experimental study on the effect of codirectional Raman gain on system's performance. AB - We have experimentally investigated the impact of codirectional Raman gains on the performance of distributed fiber Raman amplified systems. The effects of various noise sources, such as optical signal-to-noise ratio (OSNR) degradation, fiber nonlinearities and surviving channel gain variation in different Raman pumping schemes, were evaluated as a function of input power into a fiber span. For measurements, distributed Raman gain was generated by pumping the fiber span with different combinations of Raman pump power between co- and counterdirections. From the results, we found that a large counterdirectional Raman gain assisted by a small codirectional gain could improve the overall static and dynamic performance of distributed Raman amplified systems. PMID- 19546920 TI - Design of high-order all-optical temporal differentiators based on multiple-phase shifted fiber Bragg gratings. AB - A simple and general approach for designing practical all-optical (all-fiber) arbitrary-order time differentiators is introduced here for the first time. Specifically, we demonstrate that the Nth time derivative of an input optical waveform can be obtained by reflection of this waveform in a single uniform fiber Bragg grating (FBG) incorporating N &pi-phase shifts properly located along its grating profile. The general design procedure of an arbitrary-order optical time differentiator based on a multiple-phase-shifted FBG is described and numerically demonstrated for up to fourth-order time differentiation. Our simulations show that the proposed approach can provide optical operation bandwidths in the tens of-GHz regime using readily feasible FBG structures. PMID- 19546921 TI - Optical gradient flow focusing. AB - This paper describes a new method for carrying out flow cytometry, which employs optical gradient forces to guide and focus particles in the fluid flow. An elliptically shaped Gaussian beam was focused at the center of a microchannel to exert radiation pressure on suspended nanoparticles that are passing through the channel, such that these particles are guided to the center of the channel for efficient detection and sorting. To verify the efficiency of this optical gradient-flow-focusing method, we present numerical simulations of the trajectories of the nanoparticles in both electroosmotic flow (EOF) and pressure driven flow (PDF). PMID- 19546922 TI - Q-switching of a fiber laser with a single crystal photo-elastic modulator. AB - A study of using a single crystal photo-elastic modulator for active Q-switching of a fiber laser is presented. The modulator, which oscillates in a longitudinal eigenmode, was realized with LiTaO(3). This induces due to the photo-elastic effect a modulated artificial birefringence which modulates the polarization of passing light. When used together with a polarizer inside a laser cavity the laser photon life time is strongly modulated and the laser may start to emit laser pulses. We realized this with a fiber laser based on a 5m long double clad Nd-doped fiber. The pulse repetition frequency was 400 kHz and the pulse duration 300ns. PMID- 19546923 TI - Polymer microstructured fibers by one-step extrusion. AB - For the first time to our knowledge, polymer-based microstructured fibers with complex cross-sections are directly produced via melt extrusion. Two principal types of fibers were fabricated: a microstructured fiber of a single polymer with a hexagonal array of air holes and a bicomponent fiber consisting of approximately 60 coaxial rings. From the latter, strong visible iridescence was observed and is shown to exhibit a mechanochromic response. This approach, the mainstay of the textile trade for decades, offers a means of continuous high volume low-cost manufacturing of polymer (and conceivably soft-glass) fibers. For example, in the present effort, 128 coaxially microstructured fibers were fabricated simultaneously at rates exceeding 1200 m/min from industrially mainstream polymers. This approach offers an important step forward towards commoditizing microstructured fibers and open new doors for optical engineering in fashion, marking/identification, and numerous military applications. PMID- 19546924 TI - All-Optical flip-flop operation using a SOA and DFB laser diode optical feedback combination. AB - We report on the switching of an all-optical flip-flop consisting of a semiconductor optical amplifier (SOA) and a distributed feedback laser diode (DFB), bidirectionally coupled to each other. Both simulation and experimental results are presented. Switching times as low as 50ps, minimal required switch pulse energies below 1pJ and a repetition rate of 1.25GHz have been measured. Contrast ratios over 25dB have been obtained. The dependence on the pulse length and CW input power of the minimal required switch energy is investigated. PMID- 19546925 TI - Angle-resolved optical coherence tomography with sequential angular selectivity for speckle reduction. AB - We present a novel method for rapidly acquiring optical coherence tomography (OCT) images at multiple backscattering angles. By angularly compounding these images, high levels of speckle reduction were achieved. Signal-to-noise ratio (SNR) improvements of 3.4 dB were obtained from a homogeneous tissue phantom, which was in good agreement with the predictions of a statistical model of speckle that incorporated the optical parameters of the imaging system. In addition, the fast acquisition rate of the system (10 kHz A-line repetition rate) allowed angular compounding to be performed in vivo without significant motion artifacts. Speckle-reduced OCT images of human dermis show greatly improved delineation of tissue microstructure. PMID- 19546926 TI - Swept source optical coherence microscopy using a Fourier domain mode-locked laser. AB - Swept source optical coherence microscopy (OCM) enables cellular resolution en face imaging as well as integration with optical coherence tomography (OCT) cross sectional imaging. A buffered Fourier domain mode-locked (FDML) laser light source provides high speed, three dimensional imaging. Image resolutions of 1.6 microm x 8 microm (transverse x axial) with a 220 microm x 220 microm field of view and sensitivity higher than 98 dB are achieved. Three dimensional cellular imaging is demonstrated in vivo in the Xenopus laevis tadpole and ex vivo in the rat kidney and human colon. PMID- 19546927 TI - Beam homogenizers based on chirped microlens arrays. AB - Lens array arrangements are commonly used for the homogenization of highly coherent laser beams. These fly's eye condenser configurations can be used to shape almost arbitrary input intensity distributions into a top hat. Due to the periodic structure of regular arrays the output intensity distribution is modulated by equidistant sharp intensity peaks which are disturbing the homogeneity. As a new approach we apply chirped microlens arrays to the beam shaping system. These are non-regular arrays consisting of individually shaped lenses defined by a parametric description which can be derived completely from analytical functions. The advantages of the new concept and design rules are presented. PMID- 19546928 TI - Different discrete soliton states in periodic optical induced waveguide lattice. AB - Different discrete soliton states in optical waveguide lattices are studied theoretically for the different external conditions such as the biased field, lattice spacing and the position of the input pulse. It is demonstrated that the discrete solitons can be formed in an appropriate biased field and exhibit different discrete properties in different biased field. The results also indicate that lattice spacing and the position of input pulse can affect apparently the discreteness of discrete solitons. If changing the ratio of the waist width of input beam to lattice period, the discrete solitons will be excited in more or less channels. In addition, the increase of lattice intensity almost does not affect the propagation behavior of soliton. PMID- 19546929 TI - Non-Bloch plasmonic stop-band in real-metal gratings. AB - Recent studies of plasmon surface wave (PSW) propagation in short-period perfectly conducting gratings have shown formation of stop-band that are not linked to the interaction between two (counter) propagating surface waves. We study the properties of this stop-band in real metals. While for both perfectly conducting and real metals the propagation constant of PSW grows with the groove height, the stop-band in real metals appears for groove heights significantly smaller than in perfect metals. A physical explanation of the formation of the stop-band is proposed both by using a homogenisation of the corrugated layer and by analysis of the tangential electric field component. PMID- 19546931 TI - Microcavities based on multimodal interference. AB - We describe intricate cavity mode structures, that are possible in waveguide devices with two or more guided modes. The main element is interference between the scattered fields of two modes at the facets, resulting in multipole or mode cancelations. Therefore, strong coupling between the modes, such as around zero group velocity points, is advantageous to obtain high quality factors. We discuss the mechanism in three different settings: a cylindrical structure with and without negative group velocity mode, and a surface plasmon device. A general semi-analytical expression for the cavity parameters describes the phenomenon, and it is validated with extensive numerical calculations. PMID- 19546930 TI - Ultrahigh-speed optical coherence tomography imaging and visualization of the embryonic avian heart using a buffered Fourier Domain Mode Locked laser. AB - The embryonic avian heart is an important model for studying cardiac developmental biology. The mechanisms that govern the development of a four chambered heart from a peristaltic heart tube are largely unknown due in part to a lack of adequate imaging technology. Due to the small size and rapid motion of the living embryonic avian heart, an imaging system with high spatial and temporal resolution is required to study these models. Here, an optical coherence tomography (OCT) system using a buffered Fourier Domain Mode Locked (FDML) laser is applied for ultrahigh-speed non-invasive imaging of embryonic quail hearts at 100,000 axial scans per second. The high scan rate enables the acquisition of high temporal resolution 2D datasets (195 frames per second or 5.12 ms between frames) and 3D datasets (10 volumes per second). Spatio-temporal details of cardiac motion not resolvable using previous OCT technology are analyzed. Visualization and measurement techniques are developed to non-invasively observe and quantify cardiac motion throughout the brief period of systole (less than 50 msec) and diastole. This marks the first time that the preseptated embryonic avian heart has been imaged in 4D without the aid of gating and the first time it has been viewed in cross section during looping with extremely high temporal resolution, enabling the observation of morphological dynamics of the beating heart during systole. PMID- 19546933 TI - Microfabricated saturated absorption laser spectrometer. AB - We demonstrate a miniature microfabricated saturated absorption laser spectrometer. The system consists of miniature optics, a microfabricated Rb vapor cell, heaters, and a photodetector, all contained within a volume of 0.1 cm(3). Saturated absorption spectra were measured with a diode laser at 795 nm. They are comparable to signals obtained with standard table-top setups, although the rubidium vapor cell has an interior volume of only 1 mm(3). We discuss the performance and prospects for using such systems as a miniature optical wavelength reference, compatible with transportable instruments. PMID- 19546932 TI - Stabilization of counterpropagating solitons by photonic lattices. AB - We report on the stabilization of inherently unstable counterpropagating photorefractive spatial solitons by the use of one- and two-dimensional photonic lattices. We numerically investigate the dependence of the instability dynamics on period and amplitude of the lattice and present experimental verification for the dynamic stabilization of the bi-directional soliton state. PMID- 19546934 TI - Fractionalization of optical beams: II. Elegant Laguerre-Gaussian modes. AB - We apply the tools of fractional calculus to introduce new fractional-order solutions of the paraxial wave equation that smoothly connect the elegant Laguerre-Gaussian beams of integral-order. The solutions are characterized in general by two fractional indices and are obtained by fractionalizing the creation operators used to create elegant Laguerre-Gauss beams from the fundamental Gaussian beam. The physical and mathematical properties of the circular fractional beams are discussed in detail. The orbital angular momentum carried by the fractional beam is a continuous function of the angular mode index and it is not restricted to take only discrete values. PMID- 19546935 TI - Quasistatic cloaking of two-dimensional polarizable discrete systems by anomalous resonance. AB - Discrete systems of infinitely long polarizable line dipoles are considered in the quasistatic limit, interacting with a two-dimensional cloaking system consisting of a hollow plasmonic cylindrical shell. A numerical procedure is described for accurately calculating electromagnetic fields arising in the quasistatic limit, for the case when the relative permittivity of the cloaking shell has a very small imaginary part. Animations are given which illustrate cloaking of discrete systems, both for the case of induced dipoles and induced quadrupoles on the interacting particles. The simulations clarify the physical mechanism for the cloaking. PMID- 19546936 TI - Higher-band modulational instability in photonic lattices. AB - Propagation of extended Floquet-Bloch modes in the first three bands of a one dimensional photonic lattice possessing a self-defocusing saturable nonlinearity is studied experimentally and numerically on the example of waveguide arrays in lithium niobate. Discrete modulation instability is observed in all bands in the region of anomalous diffraction, whereas modes propagate stable in the normal diffraction regime. PMID- 19546937 TI - Direct detection of optical phase conjugation in a colloidal medium. AB - Degenerate four-wave mixing is demonstrated using an artificial Kerr medium and is evidenced by directly observing the phase conjugation of a vortex signal beam. The nonlinear susceptibility is produced by a refractive index grating created in a suspension of dielectric microscopic particles optically confined in the intensity grating distribution of two interfering laser beams. PMID- 19546938 TI - LDPC coded OFDM over the atmospheric turbulence channel. AB - Low-density parity-check (LDPC) coded optical orthogonal frequency division multiplexing (OFDM) is shown to significantly outperform LDPC coded on-off keying (OOK) over the atmospheric turbulence channel in terms of both coding gain and spectral efficiency. In the regime of strong turbulence at a bit-error rate of 10(-5), the coding gain improvement of the LDPC coded single-side band unclipped OFDM system with 64 sub-carriers is larger than the coding gain of the LDPC coded OOK system by 20.2 dB for quadrature-phase-shift keying (QPSK) and by 23.4 dB for binary-phase-shift keying (BPSK). PMID- 19546939 TI - Simple estimation of pulse amplitude noise and timing jitter evolution through the temporal Talbot effect. AB - We present a simple way to analytically predict the effect of the temporal Talbot self-imaging process on random amplitude noise and timing jitter in periodic optical pulse trains. The analysis is general and can be applied to any pulse shape; simulation results are in excellent agreement with the predicted values. In addition, the results clearly show that the temporal Talbot effect has an inherent property of mitigating the standard deviation of both pulse amplitude noise and timing jitter. PMID- 19546940 TI - Molded transparent photopolymers and phase shift optics for fabricating three dimensional nanostructures. AB - This paper introduces approaches that combine micro/nanomolding, or nanoimprinting, techniques with proximity optical phase mask lithographic methods to form three dimensional (3D) nanostructures in thick, transparent layers of photopolymers. The results demonstrate three strategies of this type, where molded relief structures in these photopolymers represent (i) fine (<1 microm) features that serve as the phase masks for their own exposure, (ii) coarse features (>1 microm) that are used with phase masks to provide access to large structure dimensions, and (iii) fine structures that are used together phase masks to achieve large, multilevel phase modulations. Several examples are provided, together with optical modeling of the fabrication process and the transmission properties of certain of the fabricated structures. These approaches provide capabilities in 3D fabrication that complement those of other techniques, with potential applications in photonics, microfluidics, drug delivery and other areas. PMID- 19546941 TI - Micro knife-edge optical measurement device in a silicon-on-insulator substrate. AB - The knife-edge method is a commonly used technique to characterize the optical profiles of laser beams or focused spots. In this paper, we present a micro knife edge scanner fabricated in a silicon-on-insulator substrate using the micro electromechanical-system technology. A photo detector can be fabricated in the device to allow further integration with on-chip signal conditioning circuitry. A novel backside deep reactive ion etching process is proposed to solve the residual stress effect due to the buried oxide layer. Focused optical spot profile measurement is demonstrated. PMID- 19546942 TI - Visible to infrared photoluminescence from gold nanoparticles embedded in germano silicate glass fiber. AB - Germano-silicate glass fiber containing gold nanoparticles was developed by modified chemical vapor deposition and solution doping processes. Pumping with 488 nm Argon ion laser, we firstly report on the visible to infrared photoluminescence of the gold nanoparticles embedded in the core of the germano silicate fibers. The surface plasmon resonance absorption peak at 498.4 nm and the visible to infrared photoluminescence over the range of 600 nm approximately 1560 nm were found and explained according to the interband and intraband electronic transitions of Au atoms. The averaged quantum efficiencies of the photoluminescence at 833 nm and 1536 nm were estimated to be 5.75 x 10(-8) and 2.01 x 10(-9), respectively. PMID- 19546943 TI - Resonant tunneling of surface plasmon-polaritons. AB - The tunneling of surface plasmon-polaritons (SPPs) across an interruption in the metallic film supporting them is numerically investigated in details. Both non symmetrical and symmetrical geometries are considered. A very high tunneling efficiency is calculated for the long-range surface plasmon in the symmetrical geometry, with an amplitude transmission as high as 80% over a 5 microm gap for a 40 nm thick gold film illuminated at lambda= 785 nm. The transmission is somewhat lower in the non-symmetrical geometry. The coupling between the different SPP modes (radiative and non-radiative) in that geometry is also investigated in detail. This coupling depends periodically upon the length of the gap. Overall, the results indicate that SPPs are not very sensitive to technological imperfections and can survive large waveguide interruptions. PMID- 19546945 TI - Numerical investigation of electromagnetically induced transparency in a quantum dot structure. AB - .A numerical investigation of pulse propagation in a quantum dot structure in the regime of electromagnetically induced transparency is reported. The quantum dot is described as a cone on top of a wetting layer and the calculated energy levels and dipole moments are used in an effective three-level model. Pulse propagation characteristics such as degree of slowdown, absorption, and pulse distortion are investigated with respect to their dependence on the dephasing rates and pulse width. It is seen how Rabi oscillations can seriously distort the pulse when the spectral width of the pulse becomes too large compared to the width of the EIT window. PMID- 19546944 TI - High-power fifth-harmonic generation of femtosecond pulses in the vacuum ultraviolet using a Ti:sapphire laser. AB - We demonstrate the generation of fifth-harmonic pulses at 161 nm, with an energy of up to 600 nJ and 160 fs pulse duration from a Ti:sapphire laser at 1 kHz repetition rate by four-wave difference-frequency mixing in argon-filled waveguides. The efficiency is greatly improved by coupling to higher-order transverse modes, as well as by coating the inner surface of the waveguide. A numerical model of the process yields an understanding of the main effects influencing the harmonic generation. PMID- 19546946 TI - Ultra long high resolution beam by multi-zone rotationally symmetrical complex pupil filter. AB - An ultra long high resolution beam with extension of depth of focus (DoF) in the axial direction as well as high resolution in the transverse direction has been demonstrated by a seven-zone rotationally symmetrical complex pupil filter imposed at the aperture of a focusing lens. Both amplitude and phase of the transmitted light are modulated in different zones. The scalar diffraction theory is used to optimize the zone parameters. Simulation results show that extended DoF of the beam is increased by 16 times while the spot size at the beam waist is reduced to 0.7 times. PMID- 19546947 TI - Broadband circular polarizer using stacked chiral polymer films. AB - A scattering-free broadband (approximately 120 nm bandwidth) circular polarizer is demonstrated by stacking three chiral polymer films with different pitch lengths. Using 4 x 4 matrix method, we have theoretically simulated the transmission spectra of each chiral polymer film and the three stacked films. Simulation results agree well with experiment. A broadband circular polarizer with bandwidth ranging from 400 to 736 nm can be achieved by stacking 8 such chiral polymer films together. Simulation results indicate that if a high birefringence (Deltan approximately 0.35) polymer film is employed the number of films can be reduced to three. Potential applications of these circular polarizers for liquid crystal displays, optical communications, and optical remote sensors are discussed. PMID- 19546948 TI - Three-dimensional laser microsurgery in light-sheet based microscopy (SPIM). AB - Advances in the life sciences rely on the ability to observe dynamic processes in live systems and in environments that mimic in-vivo situations. Therefore, new methodological developments have to provide environments that resemble physiologically and clinically relevant conditions as closely as possible. In this work, plasma-induced laser nanosurgery for three-dimensional sample manipulation and sample perturbation is combined with optically sectioning light sheet based fluorescence microscopy (SPIM) and applied to three-dimensional biological model systems. This means: a) working with a biological system that is not confined to essentially two dimensions like cell cultures on cover glasses, b) gaining intrinsic optical sectioning capabilities by an efficient three dimensional fluorescence imaging system, and c) using arbitrarily-shaped three dimensional ablation-patterns by a plasma-induced laser ablation system that prevent damage to surrounding tissues. Spatial levels in our biological applications range from sub-microns during delicate ablation of single microtubules over the confined disruption of cell membranes in an MDCK-cyst to the macroscopic cutting of a millimeter-sized Zebrafish caudal fin with arbitrary three-dimensional ablation patterns. Dynamic processes like laser-induced hemocyte migration can be studied with our SPIM-microscalpel in intact, live embryos. PMID- 19546949 TI - Mode counting in high-dimensional orbital angular momentum entanglement. AB - We study the high-dimensional orbital angular momentum OAM) entanglement contained in the spatial profiles of two quantum-correlated photons. For this purpose, we use a multi-mode two-photon interferometer with an image rotator in one of the interferometer arms. By measuring the two-photon visibility as a function of the image rotation angle we measure the azimuthal Schmidt number, i.e., we count the number of OAM modes involved in the entanglement; in our setup this number is tunable from 1 to 8. PMID- 19546950 TI - Design optimization of flattop interleaver and its dispersion compensation. AB - The objective of this paper is to present a general strategy for design optimization of flattop interleavers, and dispersion compensation for the interleavers, in order to achieve superior optical performance. The interleaver is formed by two multi-cavity Gire-Tournois etalons (MC-GTE) in a Michelson Interferometer (MI). An interleaver that has m cavities in one etalon and n cavities in the other is called an mn-GTE interleaver. Our optimization strategy exploits the general flattop condition and the technique of ripple equalization. Any mn-GTE interleaver may be optimized. The spectral performance can be greatly improved by the optimization process. As an illustration, we present a comprehensive analysis for a 11-GTE and a 21-GTE interleaver. The analytical expressions for flattop conditions, peak and trough positions are derived for optimization. The optimal performance of the interleavers can be controlled by the reflection coefficients and the parameters m and n. To achieve low-dispersion mn-GTE flattop interleavers, we propose to use one additional MC-GTE as a dispersion compensator to compensate for the chromatic dispersion. The analytical expressions of group delays and chromatic dispersions for an MC-GTE interleaver are derived. The optimization strategy of dispersion-ripple equalization is explained. The results show that the dispersion performance can be tailored by changing the reflection coefficients of the MC-GTE, and the dispersion and bandwidth can be enhanced by increasing the number of cavities of the MC-GTE. PMID- 19546951 TI - Quasi-CW diode-pumped self-starting adaptive laser with self-Q-switched output. AB - An investigation is made into a quasi-CW (QCW) diode-pumped holographic adaptive laser utilising an ultra high gain (approximately 10(4)) Nd:YVO(4) bounce amplifier. The laser produces pulses at 1064 nm with energy approximately 0.6 mJ, duration <3 ns and peak power approximately 200 kW, with high stability, via self Q-switching effects due to the transient dynamics of the writing and replay of the gain hologram for each pump pulse. The system produces a near-diffraction limited output with M(2)<1.3 and operates with a single longitudinal mode. In a further adaptive laser configuration, the output was amplified to obtain pulses of approximately 5.6 mJ energy, approximately 7 ns duration and approximately 1 MW peak power. The output spatial quality is also M(2)<1.3 with SLM operation. Up to 2.9 mJ pulse energy of frequency doubled green (532 nm) radiation is obtained, using an LBO crystal, representing approximately 61% conversion efficiency. This work shows that QCW diode-pumped self-adaptive holographic lasers can provide a useful source of high peak power, short duration pulses with excellent spatial quality and narrow linewidth spectrum. PMID- 19546952 TI - High energy, single-mode, narrow-linewidth fiber laser source using stimulated Brillouin scattering beam cleanup. AB - We propose an original nonlinear beam cleaning fiber laser architecture to obtain high energy pulses with a good beam quality and a narrow linewidth. The output beam of a large core Er:Yb co-doped multimode fiber amplifier (M(2) approximately 6, 220 microJ) is converted into a near diffraction limited beam (M(2) = 1.6) through a stimulated Brillouin scattering injection seeded beam cleanup process. We report in this experiment a multimode to single mode conversion efficiency of 50% while preserving the master oscillator linewidth. PMID- 19546953 TI - Dielectric microsphere manipulation and chain assembly by counter-propagating waves in a channel waveguide. AB - We study the formation and the propulsion properties of chains of dielectric microspheres in the evanescent field of a channel waveguide made by Cs(+) ion exchange. Particle chains are shown to move faster than single particles. We exploit counter-propagating waves for axial positioning of single and chains of microspheres. The particles can be propelled back and forth at will, and trapped at a given point for several minutes. We demonstrate that this technique can also be used to assemble a long, one-particle wide, chain. PMID- 19546954 TI - Compact, high average power, fiber-pumped terahertz source for active real-time imaging of concealed objects. AB - We have modeled and demonstrated a scalable, compact, fiber-pumped terahertz source based on difference frequency mixing (DFM) in zinc germanium phosphide (ZGP) capable of producing high average and peak-power pulses. Currently, our terahertz source produces 2 mW of average THz power and >40 W of peak power with sub-nanosecond pulses at a repetition rate of 100 kHz in the range of 2-3 THz without cryogenic cooling or ultra-fast optics. This high average power laser based terahertz output enables the real-time imaging of concealed objects using an off-the-shelf uncooled microbolometer focal-plane array. With this THz system, we have imaged objects obscured inside in a common shipping envelope, demonstrating the potential of compact laser-based terahertz sources for use in security screening applications. PMID- 19546955 TI - Double-control quantum interferences in a four-level atomic system. AB - A new scheme is suggested to manipulate the probe transitions (and hence the optical properties of atomic vapors) via double-control destructive and constructive quantum interferences. The influence of phase coherence between the two control transitions on the probe transition is also studied. The most remarkable feature of the present scheme is that the optical properties (absorption, transparency and dispersion) of an atomic system can be manipulated using this double-control multi-pathway interferences (multiple routes to excitation). It is also shown that a four-level system will exhibit a two-level resonant absorption because the two control levels (driven by the two control fields) form a dark state (and hence a destructive quantum interference occurs between the two control transitions). However, the present four-level system will exhibit electromagnetically induced transparency to the probe field when the three lower levels (including the probe level and the two control levels) form a three-level dark state. The present scenario has potential applications in new devices (e.g. logic gates and sensitive optical switches) and new techniques (e.g. quantum coherent information storage). PMID- 19546956 TI - Symmetric 3 x 3 optical coupler using three parallel long-period fiber gratings. AB - We propose a new technique of realizing a symmetric 3 x 3 optical coupler, which is based on evanescent-field coupling among three identical parallel long-period fiber gratings placed side-by-side. Using three 32-mm-long over-coupled long period fiber gratings embedded in a medium with a proper refractive index, we achieved equal power outputs from the three fibers at the resonance wavelength of the gratings with a total loss of about 1.4 dB when light was launched into any one of the fibers. PMID- 19546957 TI - Pulse compression and modelocking by using TPA in silicon waveguides. AB - We demonstrate a novel broadband pulse compression and modelocking scheme by using two-photon absorption in silicon waveguides. Experimentally we obtain greater than 20 fold pulse compression and 200 ps modelocked pulses. The free carrier lifetime and the width of the modulation signal are found to be two critical parameters affecting the output pulse width. Theoretical calculations indicate that optical pulses of less than 20 ps width are achievable by using the same technique. PMID- 19546958 TI - Biphoton interference with a quantum dot entangled light source. AB - We demonstrate optical interferometry beyond the limits imposed by the photon wavelength using 'triggered' entangled photon pairs from a semiconductor quantum dot. Interference fringes of the entangled biphoton state reveals a periodicity half of that obtained with the single photon, and much less than that of the pump laser. High fringe visibility indicates that biphoton interference is less sensitive to decoherence than interference of two sequential single photons. The results suggest that quantum interferometry may be possible using a semiconductor LED-like device. PMID- 19546959 TI - General analysis of group velocity effects in collinear optical parametric amplifiers and generators. AB - Group velocity mismatch (GVM) is a major concern in the design of optical parametric amplifiers (OPAs) and generators (OPGs) for pulses shorter than a few picoseconds. By simplifying the coupled propagation equations and exploiting their scaling properties, the number of free parameters for a collinear OPA is reduced to a level where the parameter space can be studied systematically by simulations. The resulting set of figures show the combinations of material parameters and pulse lengths for which high performance can be achieved, and they can serve as a basis for a design. PMID- 19546960 TI - Spatially extended FCS for visualizing and quantifying high-speed multiphase flows in microchannels. AB - We report the development of spatially extended fluorescence correlation spectroscopy for visualizing and quantifying multiphase flows in microchannels. We employ simultaneous detection with a high-speed camera across the width of the channel, enabling investigation of the dynamics of the flow at short time scales. We take advantage of the flow to scan the sample past the fixed illumination, capturing frames up to 100 KHz. At these rates, we can resolve the motion of sub micron particles at velocities up to the order of 1 cm/s. We visualize flows with kymographs and quantify velocity profiles by cross-correlations within the focal volume. We demonstrate the efficacy of our approach by measuring the depth resolved velocity profile of suspensions of sub-micron diameter silica particles flowing up to 1.5 mm/s. PMID- 19546961 TI - Wide band negative magnetic permeability materials (NMPM) with composite metalsemiconductor structures based on the Drude model, and applications to negative-refractive index (NIM). AB - Composite structures based on metal open rings and thin wires are well established, for obtaining efficient negative index materials (NIM), acting as metamaterials in the long wavelength regime. The main losses are due both to metal absorption and to the inner electric resistance of metals; to overcome this latter loss we propose a new metal-semiconductor structure dimensioned by direct synthesis method, which offers an almost perfect Drude-like effective magnetic permeability. The choice of particular semiconductor components allows to get a negative resistance for the current induced by the electromagnetic field, which cancels that of the metal but puts a limit to the spectral response of the metamaterial. We consider some parasite effects, such as bianisotropy and incorrect values of structural parameters, to see limitations and features of this new NIM technology. PMID- 19546962 TI - Infrared emission and energy transfer in Tm(3+), Tm(3+)-Ho(3+) and Tm(3+)-Yb(3+) doped tellurite fibre. AB - The infrared emission spectra and decay lifetimes of Tm(3+)-doped and Tm(3+) Ho(3+), Tm(3+)-Yb(3+) co-doped tellurite fibres were measured using 808 nm and 978 nm diode laser pump sources in the range 1.35 mum to 2.2 mum. The spectra were compared with varying fibre lengths and core diameters. Tm(3+)-doped fibre shows strong emission at ~1.8 mum and when co-doped with Ho(3+), energy transfer results in strong Ho(3+) fluorescence at ~2.0 mum. These fibres show promise for compact mid-IR fibre laser sources. PMID- 19546963 TI - Metallic wave-impedance matching layers for broadband terahertz optical systems. AB - We examine the potential of ultra-thin metallic layers for broadband wave impedance matching in the terahertz frequency range. The metallic layer is modeled using Fresnel formulae for stratified optical medium. Experimental data for chromium and indium-tin-oxide layers, measured using time-domain terahertz spectroscopy over the frequency range 0.4 - 4.5 THz, are compared with theoretical results. PMID- 19546964 TI - Antiresonant-ring Kerr spectroscopy. AB - We demonstrate a new scheme for measuring different tensor elements of the optical Kerr effect response. A dual-ring, polarization-dependent Sagnac interferometer is used to create two copropagating probe pulses that arrive at the sample at different times but that reach the detector simultaneously and collinearly. The tensor element of the response that is measured is determined by the polarization of the pump pulse. By controlling the relative timing of the probe pulses it is also possible to perform optical subtraction of two different tensor elements of the response at two different times, a strategy that can be used to enhance or suppress particular contributions to the OKE response. PMID- 19546966 TI - Surface plasmon polariton beam focusing with parabolic nanoparticle chains. AB - We report on the focusing of surface plasmon polariton (SPP) beams with parabolic chains of gold nanoparticles fabricated on thin gold films. SPP focusing with different parabolic chains is investigated in the wavelength range of 700-860 nm, both experimentally and theoretically. Mapping of SPP fields is accomplished by making use of leakage radiation microscopy, demonstrating robust and efficient SPP focusing into submicron spots. Numerical simulations based on the Green's tensor formalism show very good agreement with the experimental results, suggesting the usage of elliptical corrections for parabolic structures to improve their focusing of slightly divergent SPP beams. PMID- 19546965 TI - Efficient coupling into slow-light photonic crystal channel guides using photonic crystal tapers. AB - Photonic crystal tapers have been designed for coupling of light from ridge waveguides into low group velocity photonic crystal channel guides. The coupling efficiency is increased from 3 % (case of butt-coupling) to 97 % for frequencies in the band-edge region, corresponding to a group index close to 100, as predicted using 2D finite-difference time-domain simulations. PMID- 19546967 TI - Molecular specific optoacoustic imaging with plasmonic nanoparticles. AB - Gold nanoparticles functionalized with antibodies can specifically bind to molecular biomarkers such as epithelial growth factor receptor (EGFR). The molecule specific nature of the antibody-functionalized gold nanoparticles forms the basis for the developed optoacoustic imaging technique to detect cancer at an asymptotic stage. Optoacoustic imaging was performed with 532 nm and 680 nm pulsed laser irradiation on three-dimensional tissue phantoms prepared using a human keratinocyte cell line. The results of our study demonstrate that the combination of anti-EGFR gold ioconjugates and optoacoustic imaging can allow highly sensitive and selective detection of human epithelial cancer cells. PMID- 19546968 TI - Calibration of scattering and absorption properties of a liquid diffusive medium at NIR wavelengths. Time-resolved method. AB - In this paper, a general method to calibrate the absorption coefficient of an absorber and the reduced scattering coefficient of a liquid diffusive medium, based on time-resolved measurements, is reported. An exhaustive analysis of the error sources affecting the estimation is also performed. The method has been applied with a state-of-the-art time-resolved instrumentation to determine the intrinsic absorption coefficient of Indian ink and the reduced scattering coefficient of Intralipid-20%, with a standard error smaller than 1% and 2%, respectively. Finally, the results have been compared to those retrieved for the same compounds by applying a continuous wave method recently published, obtaining an agreement within the error bars. This fact represents a cross validation of the two independent calibration methods. PMID- 19546969 TI - Influence of surface termination on negative reflection by photonic crystals. AB - When a wave impinges obliquely to the interface of a Photonic Crystal (PhC), the wave can be completely reflected in counter-propagating direction instead of the usually expected specular direction. However the beam is totally specularly reflected with a simple modification of the surface termination. The analysis of the time average Poynting vector evidences that PhC termination modifies the energy flow and determines the reflection properties. PMID- 19546970 TI - Anomalous Faraday effect of a system with extraordinary optical transmittance. AB - It is shown theoretically that the Faraday rotation becomes anomalously large and exhibits extraordinary behavior near the frequencies of the extraordinary optical transmittance through optically thick perforated metal film with holes filled with a magneto-optically active material. This phenomenon is explained as result of strong confinement of the evanescent electromagnetic field within magnetic material, which occurs due to excitation of the coupled plasmon-polaritons on the opposite surfaces of the film. PMID- 19546971 TI - A passively-modelocked, Yb-doped, figure-eight, fiber laser utilizing anomalous dispersion higher-order-mode fiber. AB - Modelocking in an Yb-doped figure-eight fiber laser is demonstrated utilizing anomalous dispersion from an LP(02) higher-order-mode fiber for dispersion management. Outside the laser cavity, the pulses were re-compressed to 95 fs using a second HOM module, the shortest demonstrated pulses to date from an Yb doped figure-eight fiber laser. Operation of the laser with HOM fiber in the cavity is compared to an Yb figure-eight laser that utilizes all-normal dispersion fibers. PMID- 19546972 TI - Field and dispersion properties of subwavelength-diameter hollow optical fiber. AB - We have investigated the basic properties of subwavelength- diameter hollow optical fiber with exact solutions of Maxwell's equations. The characteristics of modal field and waveguide dispersion have been studied. It shows that the subwavelength-diameter hollow optical fibers have interesting properties, such as enhanced evanescent field, local enhanced intensity in the hollow core and large waveguide dispersion that are very promising for many miniaturized high performance and novel photonic devices. PMID- 19546973 TI - Transparent volumetric three-dimensional image display based on the luminescence of a spinning sheet with dissolved Lanthanide(III) complexes. AB - We have developed a new type of transparent volumetric three-dimensional (3D) image display in which a thin photopolymer sheet containing Lanthanide(III) complexes is used as a rotational screen. The Lanthanide(III) complexes used in our system are Eu(TTA)(3) Phen, designed for achieving red luminescence (615nm) for an excitation light of 395 nm. An arbitrary luminous point (voxel) is identified by controlling the excitation laser beam direction in synchronization with the photopolymer sheet rotation. The full colorization of the proposed volumetric 3D image display can be realized by using, for example, Eu(TTA)(3) Phen, Tb(ACAC)(3) Phen, and Coumarin 337, simultaneously. PMID- 19546974 TI - Activity of the human visual cortex measured non-invasively by diffusing-wave spectroscopy. AB - Activity of the human visual cortex, elicited by steady-state flickering at 8Hz, is non-invasively probed by multi-speckle diffusingwave spectroscopy (DWS). Parallel detection of the intensity fluctuations of statistically equivalent, but independent speckles allows to resolve stimulation-induced changes in the field autocorrelation of multiply scattered light of less than 2%. In a group of 9 healthy subjects we find a faster decay of the field autocorrelation function during the stimulation periods for data measured with a long-distance probe (30mm source-receiver distance) at 2 positions over the occipital cortex (t-test: t(8) = -2.672, p = 0.028 < 0.05 for position 1, t(8) = -2.874, p = 0.021 < 0.05 for position 2). In contrast, no statistically significant change is seen when a short-distance probe (16mm source-receiver distance) is used (t-test: t(8) = 2.043, p = 0.075 > 0.05 for position 1, t(8) = -2.146, p = 0.064 > 0.05 for position 2). The enhanced dynamics observed with DWS is positively correlated with the functional increase of blood volume in the visual cortex, while the heartbeat rate is not affected by stimulation. Our results indicate that the DWS signal from the visual cortex is governed by the regional cerebral blood flow velocity. PMID- 19546975 TI - Imaging interferometric microscopy-approaching the linear systems limits of optical resolution. AB - The linear systems optical resolution limit is a dense grating pattern at a lambda/2 pitch or a critical dimension (resolution) of lambda/4. However, conventional microscopy provides a (Rayleigh) resolution of only ~ 0.6lambda/NA, approaching lambda/1.67 as NA ?lambda1. A synthetic aperture approach to reaching the lambda/4 linear-systems limit, extending previous developments in imaginginterferometric microscopy, is presented. Resolution of non-periodic 180 nm features using 633-nm illumination (lambda/3.52) and of a 170-nm grating (lambda/3.72) is demonstrated. These results are achieved with a 0.4-NA optical system and retain the working distance, field-of-view, and depth-of-field advantages of low-NA systems while approaching ultimate resolution limits. PMID- 19546977 TI - Light extraction analysis of GaN-based light-emitting diodes with surface texture and/or patterned substrate. AB - Light extraction analysis of GaN-based light-emitting diodes (LEDs) with Monte Carlo ray tracing is presented. To obtain high light extraction efficiency, periodic structures introduced on the top surface and/or on the substrate of various types of LED are simulated, including wire bonding, flip chip and Thin GaN. Micro pyramid array with an apex angle from 20o to 70o is shown to effectively improve the light extraction efficiency. In addition, for an LED encapsulated within an epoxy lens, the patterned substrate with pyramid array is found to be a more effective way to increase light extraction efficiency than the surface texture. PMID- 19546976 TI - Design of a zoom lens without motorized optical elements. AB - A novel design of a zoom lens system without motorized movements is proposed. The lens system consists of a fixed lens and two double-liquid variable-focus lenses. The liquid lenses, made out of two immiscible liquids, are based on the principle of electrowetting: an effect controlling the wetting properties of a liquid on a solid by modifying the applied voltage at the solid-liquid interface. The structure and principle of the lens system are introduced in this paper. Detailed calculations and simulation examples are presented to show that this zoom lens system appears viable as the next-generation zoom lens. PMID- 19546978 TI - Pulse dynamics in mode-locked lasers: relaxation oscillations and frequency pulling. AB - A theoretical description of the pulse dynamics in a modelocked laser including gain dynamics is developed. Relaxation oscillations and frequency pulling are predicted that influence the pulse parameters. Experimental observations of the response of a mode-locked Ti:sapphire laser to an abrupt change in the pump power confirm that the predicted behavior occurs. These results provide a framework for understanding the effects of noise on the spectrum of the laser. PMID- 19546979 TI - Photonic band-gap fiber gas cell fabricated using femtosecond micromachining. AB - Femtosecond laser drilling is used to produce a variablepressure fiber gas cell. Tightly focused laser pulses are used to produce micrometer-diameter radial channels in a hollow-core photonic band-gap fiber (HC-PBGF), and through these microchannels the core of the fiber is filled with a gas. The fiber cell is formed by fusion splicing and sealing the ends of the HC-PBGF to standard step index fiber. As a demonstration, acetylene is introduced into an evacuated fiber at multiple backing pressures and spectra are measured. PMID- 19546980 TI - Three-dimensional in vivo fluorescence diffuse optical tomography of breast cancer in humans. AB - We present three-dimensional (3D) in vivo images of human breast cancer based on fluorescence diffuse optical tomography (FDOT). To our knowledge, this work represents the first reported 3D fluorescence tomography of human breast cancer in vivo. In our protocol, the fluorophore Indocyanine Green (ICG) is injected intravenously. Fluorescence excitation and detection are accomplished in the soft compression, parallel-plane, transmission geometry using laser sources at 786 nm and spectrally filtered CCD detection. Phantom and in vivo studies confirm the signals are due to ICG fluorescence, rather than tissue autofluorescence and excitation light leakage. Fluorescence images of breast tumors were in good agreement with those of MRI, and with DOT based on endogenous contrast. Tumorto- normal tissue contrast based on ICG fluorescence was two-to-four-fold higher than contrast based on hemoglobin and scattering parameters. In total the measurements demonstrate that FDOT of breast cancer is feasible and promising. PMID- 19546981 TI - Tunable dispersion-tolerant picosecond flat-top waveform generation using an optical differentiator. AB - We study the influence of dispersive propagation on picosecond flat-top pulses, which are generated using long period fiber grating (LPG)-based optical differentiators. We suggest an extremely simple scheme to compensate for the dispersion-induced flat-top pulse distortion; this scheme is based on proper tuning the LPG coupling strength. As this coupling strength may be changed via LPG axial straining, the demonstrated device can be tuned to compensate for different levels of the dispersion in a very easy and straightforward fashion. This allows for very fine flat-top pulse shape adjustment, even after propagation through a relatively long section of dispersive optical fiber. In the experimental demonstration reported here, the dispersion tolerance of 1.8-ps flat top pulses propagating through a standard telecom fiber (SMF-28) was increased from approximately 2 m to approximately 18 m, giving a 9-fold improvement. PMID- 19546982 TI - Point defect engineered Si sub-bandgap light-emitting diode. AB - We present a novel approach to enhance light emission in Si and demonstrate a sub bandgap light emitting diode based on the introduction of point defects that enhance the radiative recombination rate. Ion implantation, pulsed laser melting and rapid thermal annealing were used to create a diode containing a self interstitial-rich optically active region from which the zero-phonon emission line at 1218 nm originates. PMID- 19546983 TI - Retrieval of atmospheric particles optical properties by combining ground-based and spaceborne lidar elastic scattering profiles. AB - A simple algorithm is derived to retrieve the aerosol backscattering and extinction vertical profiles from simultaneously detected ground and space elastic lidar signals, without any a priori hypothesis on aerosol particles properties. This technique can be applied at any wavelength whenever two "counter looking" lidars are available and the atmosphere can be considered horizontally homogeneous in a spatial scale of the order of the distance between the two lidar beams. To test the accuracy of the algorithm a numerical simulation has been performed. Moreover, it has been applied in a real case to level 1 products from CALIPSO. PMID- 19546984 TI - Electrically pumped InP-based microdisk lasers integrated with a nanophotonic silicon-on-insulator waveguide circuit. AB - A compact, electrically driven light source integrated on silicon is a key component for large-scale integration of electronic and photonic integrated circuits. Here we demonstrate electrically injected continuous-wave lasing in InP based microdisk lasers coupled to a sub-micron silicon wire waveguide, fabricated through heterogeneous integration of InP on silicon-on-insulator (SOI). The InP based microdisk has a diameter of 7.5 mum and a thickness of 1 mum. A tunnel junction was incorporated to efficiently contact the p-side of the pn-junction. The laser emits at 1.6 mum, with a threshold current as low as 0.5 mA under continuous-wave operation at room temperature, and a threshold voltage of 1.65 V. The SOI-coupled laser slope efficiency was estimated to be 30 muW/mA, with a maximum unidirectional output power of 10 muW. PMID- 19546985 TI - Spectroscopy and concentration quenching of the infrared emissions in Tm(3+) doped TeO(2)-TiO(2)-Nb(2)O(5) glass. AB - In this work, we report the optical properties of Tm(3+) ions in tellurite glasses (TeO(2)-TiO(2)-Nb(2)O(5)) for different Tm(3+) concentrations ranging between 0.05 and 1 wt%. Judd-Ofelt intensity parameters have been determined to calculate the radiative transition probabilities and radiative lifetimes of excited states. The stimulated emission cross-sections of the infrared emissions at 1487 nm and 1800 nm have been determined from the line shape of the emission spectra and the lifetimes of levels (3)H(4) and (3)F(4) respectively. The emission spectra obtained under 793 nm excitation reveal the existence of energy transfer via cross-relaxation among Tm(3+) ions. As a result, the intensity of the infrared (3)H(4)?(3)F(4) emission at 1487 nm decreases in relation to the one at 1800 nm, as concentration increases. The non-exponential character of the decays from the (3)H(4) level with increasing concentration indicates the presence of a dipole-dipole quenching process assisted by energy migration. The self-quenching of the (3)F(4)?(3)H(6) emission at 1800 nm can be attributed to limited diffusion within the active centers. PMID- 19546986 TI - Plasmonic transmission lines: from micro to nano scale with lambda/4 impedance matching. AB - The downscaling of conventional RF transmission lines methodologies to subwavelength plasmonic circuits is discussed and demonstrated for a lambda/4 transformer impedance matching. The nano-size transformer, matching between 0.5mum and 50nm wide plasmonic transmission lines, enhances the coupling efficiency by more than 285% compared to the direct ("end fire") coupling - i.e. harvesting more than 86% of total incident power. The influence of the transverse resonances induced by the metal claddings of the input transmission line on the light harvesting is discussed as well. PMID- 19546987 TI - Optical resonators with ten million finesse. AB - We report a significant improvement of the finesse and quality factor of a calcium fluoride whispering gallery mode optical resonator achieved with iterative thermal annealing. The initial and final values of the resonator finesse are F(i) approximately 5 x 10(5) and F(f) >/= 10(7) respectively. The measurements are conducted at room temperature in the laboratory atmosphere. PMID- 19546988 TI - Fabrication of a close-packed hemispherical submicron lens array and its application in photolithography. AB - The fabrication and characteristics of close-packed lens arrays with a feature size close to the optical diffraction limit are presented in this study. By controlling the size of the submicron nickel rods and the time for reactive dry etching, the hemispherical lens array with a submicron period is made directly on a borosilicate glass. Finite-difference time-domain calculations and optical near field measurements show that such a lens array can generate a subwavelength optical spot array near the glass surface. Moreover, the spot array periodically appears in the propagation direction. Using this novel optical property, we propose a photolithographic method for the mass-production of multilayer hexagonal structures with a period of 500nm. PMID- 19546989 TI - External stimulus driven variable-step grating in a nematic elastomer. AB - We report on the creation of micro-patterns in an oriented nematic elastomer (an artificial muscle material) by photopolymerization of surface aligned nematic liquid crystal monomers. We demonstrate that microscopic techniques are able to create accurate patterns in rubber-like liquid crystal materials. Two approaches, based on one and two-photon excitations respectively, are implemented using a microscope-based setup. Due to its high spatial selectivity, the two-photon excitation mode yields finer patterns. Benefitting from the intrinsic, thermally induced, contractile properties of the material, gratings with variable steps in response to temperature changes were fabricated. PMID- 19546990 TI - Quantum limit of optimum four-level ASK signals with direct detection optically preamplified receivers. AB - We analytically obtain the exact fundamental limit of 4-level amplitude shifted keying formats (4ASK) with direct detection optically preamplified receivers. The optimum multilevel spacing and the corresponding decision thresholds, which depend both on the signal to noise ratio and optical bandwidth, are obtained numerically considering the Chi-squared distribution of each level. The quantum limit under the optimum level spacing is 127.5 photons/bit, which is about 0.2 dB smaller than the results by the Gaussian approximation. Over a broad range of the signal to noise ratio and the optical bandwidth, we have found that not only the bit error rate but also the optimum level spacing are well predicted by Gaussian method, although the three decision thresholds are all underestimated. PMID- 19546991 TI - Heisenberg limited Sagnac interferometry. AB - We show how the entangled photons produced in parametric down conversion can be used to improve the sensitivity of a Sagnac interferometer. Two-photon and four photon coincidences increases the sensitivity by a factor of two and four respectively. Our results apply to sources with arbitrary pumping and squeezing parameters. PMID- 19546992 TI - Iterative algorithm for three flat test. AB - Absolute planarity measurements by interferometry are classically made using three flats, compared two by two in the course of four or more tests. Data reduction is performed with various analytical methods. Here we present instead a data processing algorithm that converges to solution numerically by iteration. Examples are presented both on synthetic interferograms and on experimental data. High accuracy and versatility of the approach are demonstrated. PMID- 19546993 TI - Free spectral range optimization of return-to-zero differential phase shift keyed demodulation in the presence of chromatic dispersion. AB - Optical differential phase shift keying is normally demodulated in a delay-line interferometer with a 1-bit delay such that the free-spectral-range of the demodulator is equal to the transmitted bitrate. We show using Karkunen-Loeve expansion simulation that free-spectral-range optimization leads to increased chromatic dispersion tolerances. The optimized delay inversely scales with the amount of chromatic dispersion such that a delay slightly shorter than the bit period increases tolerances with no adverse effect on the polarization-mode dispersion tolerance or frequency offset penalty at the receiver. PMID- 19546994 TI - Real-time measurement of internal stress of dental tissue using holography. AB - We describe a real-time holographic technique used to observe dental contraction due to photo-polymerization of dental filling during LED lamp illumination. An off-axis setup was used, with wet in-situ processing of the holographic plate, and consequent recording of interference fringes using CCD camera. Finite elements method was used to calculate internal stress of dental tissue, corresponding to experimentally measured deformation. A technique enables selection of preferred illumination method with reduced polymerization contraction. As a consequence, durability of dental filling might be significantly improved. PMID- 19546995 TI - Combination of optical and electronic logic gates for error correction in multipath differential demodulation. AB - We present an optical multipath error correction technique for differentially encoded modulation formats such as differential-phase-shift-keying (DPSK) and differential polarization shift keying (DPolSK) for fiber-based and free-space communication. This multipath error correction method combines optical and electronic logic gates. The scheme can easily be implemented using commercially available interferometers and high speed logic gates and does not require any data overhead therefore does not affect the effective bandwidth of the transmitted data. It is not merely compatible but also complementary to error correction codes commonly used in optical transmission systems such as forward error-correction (FEC). The technique consists of separating the demodulation at the receiver in multiple paths. Each path consists of a Mach-Zehnder interferometer with a different integer bit delay used in each path. Some basic logic operations follow and the three paths are compared using a simple majority vote algorithm. Experimental results show that the scheme improves receiver sensitivity by 1.5 dB at BER of 10(-3),in back-to-back configuration. Numerical results indicate a 1.6 dB improvement in the presence of Chromatic Dispersion for a 25% increase in tolerance for a 3dB penalty from +/-1220 ps/nm to +/-1520 ps/nm. and a 0.35 dB improvement for back-to-back operation. PMID- 19546996 TI - Femtosecond absorption dynamics in glass-metal nanocomposites. AB - We report strong optical nonlinearity of glasses embedded with copper and silver nanoparticles. In pump-probe experiments with copper-doped glasses, the observed absorption bleaching with picosecond relaxation time is as high as 22%. Transmission femtosecond measurementsreveal the reverse saturable absorption with nonlinear absorption coefficient of 10(-10)cm/W in both copper- and silver-doped nanocomposites. PMID- 19546997 TI - Demonstration of slot-waveguide structures on silicon nitride / silicon oxide platform. AB - We report on the first demonstration of guiding light in vertical slot-waveguides on silicon nitride/silicon oxide material system. Integrated ring resonators and Fabry-Perot cavities have been fabricated and characterized in order to determine optical features of the slot-waveguides. Group index behavior evidences guiding and confinement in the low-index slot region at O-band (1260-1370nm) telecommunication wavelengths. Propagation losses of <20 dB/cm have been measured for the transverse-electric mode of the slot-waveguides. PMID- 19546998 TI - Backscattering of metallic microstructures with small defects located on flat substrates. AB - Micron-sized structures on flat substrates supporting submicron defects are analyzed by means of a parameter based on integrated backscattering calculations. This analysis is performed for different particle and defect sizes, optical properties and for two different configurations (defect on the microstructure or on the substrate). Calculations in the far field are complemented by some near field results. It is shown that information about the defect presence, size and optical properties) can be obtained from the proposed backscattering parameter. PMID- 19546999 TI - Photo-responsive properties and heating-induced surface relief patterns from azobenzene-doped GeO(2/gamma-)glycidoxypropyltrimethoxysilane organic-inorganic hybrid films. AB - GeO(2/ gamma-)glycidoxypropyltrimethoxysilane organic-inorganic hybrid optical waveguide films, which contain azobenzene small molecular groups, have been prepared by combining a low temperature sol-gel process with a spin-coating technique. The azobenzene in hybrid films can undergo trans-cis-trans photoisomerization efficiently by a photoirradiation with UV light. It is also found interestingly that surface relief patterns can be heat-induced on such azobenzene-containing small molecular hybrid films, but can be erased by further heating the hybrid film and no permanent damage of the hybrid film is observed. The refractive index, thickness, and morphologic properties of the hybrid films have also been investigated by a prism coupling technique and atomic force microscopy. These results indicate that the azodye-doped hybrid films are promising candidates for integrated optics and photonic applications, which allow directly integrating on the same chip waveguide device with the optical data storage and optical switching devices. PMID- 19547000 TI - In-service monitoring of 16 port x 32 wavelength bi-directional WDM-PON systems with a tunable, coded optical time domain reflectometry. AB - We propose a multi-port, multi-wavelength supervisory system for the in-service transmission line monitoring of a bidirectional WDM-PON system. Identifying unique requirements for the performance monitoring of a real field WDM-PON system, we define the architecture for the supervisory system and utilize the most up-to-date technologies (Simplex coding, tunable source, and optical switches) to demonstrate a successful interrogation of a transmission line up to 16 ports x 32 nodes (512 user) capacity. Monitoring of individual branch traces up to 60 km was achieved with the application of a 127-bit simplex code corresponding to a 7.5dB SNR coding gain. In-service transmission experiments showed negligible penalty from the monitoring system to the transmission signal quality, at a 2.5Gbps / 125Mbps (down / up stream) data rate. PMID- 19547001 TI - Multi-photon absorption upconversion luminescence of a Tb(3+)-doped glass excited by an infrared femtosecond laser. AB - In this paper we presents the near infrared to visible upconversion luminescence in a Tb(3+)-doped ZnO-B(2)O(3)-SiO(2) glass excited with 800nm femtosecond laser irradiation. The upconversion luminescence is attributed to (5)D(4) to (7)F(j) (j=3, 4, 5, 6) transitions of Tb(3+).The relationship between upconversion luminescence intensity and the pump power indicates that a three-photon simultaneous absorption process is dominant in this upconversion luminescence. The calculated value of the three-photon absorption cross section sigma(3) of the glass is 1.832x10(-81)cm(6)6s(2). Also, three-dimensional display is demonstrated based on the multiphoton absorption upconversion luminescence for the first time. PMID- 19547002 TI - All-fiber similariton laser at 1 mum without dispersion compensation. AB - We report on an all-fiber femtosecond Ytterbium laser without dispersion compensation operating in the similariton pulse regime. The oscillator was mode locked with a saturable Bragg reflector along with nonlinear polarization evolution. A pulse energy of 0.8 nJ was achieved with a pulse duration of 10 ps. The pulses could be externally dechirped to 627 fs which was within 8.7% of the Fourier transform limited pulse duration. The ealized oscillator combines compact size, high stability and alignment-free operation. PMID- 19547003 TI - Two-dimensional high-precision fiber waveguide arrays for coherent light propagation. AB - Fiber waveguide arrays can be applied as a very useful tool for the investigation of effects in discrete optics. The observation of coherent propagation in such discrete waveguide arrays requires, however, high structural precision and great material homogeneity. The fabrication of such a fiber array with close tolerances compared to conventional fiber technology is discussed. Linear propagation effects are modeled for an ideal fiber waveguide array and are compared with experimental results. The good agreement of these results with each other indicates the applicability of such fiber waveguide arrays in studying linear and non-linear properties in discrete optics. PMID- 19547004 TI - Fabrication and characterization of microlens arrays using a cantilever-based spotter. AB - We present a quantitative study on the fabrication of microlenses using a low cost polymer dispending technique. Our method is based on the use of a silicon micro-cantilever robotized spotter system. We first give a detailed description of the technique. In a second part, the fabricated microlenses are fully characterized by means of SEM (Scanning Electron Microscope), AFM (Atomic Force Microscopy) non contact optical profilometry and Mach-Zehnder interferometry. Diameters in the range [25-130mum] are obtained with an average surface roughness of 2.02nm. Curvature radii, focal lengths as well as aberrations are also measured for the first time: the fabricated microlenses present focal lengths in the range [55-181mum] and exhibit high optical quality only limited by diffraction behaviour with RMS aberration lower than lambda/14. PMID- 19547005 TI - Investigation of coupled third harmonic generation in one-dimensional defective nonlinear photonic crystals. AB - One-dimensional defective photonic crystals containing nonlinear material for coupled third harmonic generation (CTHG) have been designed. The general solution of the CTHG in such structure has been derived. The wavelengths of the fundamental wave (FW), second harmonic generation (SHG), and CTHG have been designed to lie at the defect states in different photonic band gaps by employing the simulated annealing (SA) method. Due to the strong location, low group velocity, and spatial phase locking, the conversion efficiencies of the SHG and CTHG have been greatly enhanced. Designed structure for multiple frequencies CTHG is also demonstrated. PMID- 19547006 TI - Passive and active optical bit-pattern recognition structures for multiwavelength optical packet switching networks. AB - Next generation High-Speed optical packet switching networks require components capable of recognising the optical header to enable on-the-fly accurate switching of incoming data packets to their destinations. This paper experimentally demonstrates a comparison between two different optical header recognition structures; A passive structure based on the use of Fiber Bragg Gratings (FBGs), whereas the active structure employs Opto-VLSI processors that synthesise dynamic wavelength profile through digital phase holograms. The structures are experimentally demonstrated at 10Gbps. Performance comparison between the two structures is also discussed. These optical header recognition structures are attractive for multiwavelength optical network and applications. PMID- 19547007 TI - Narrowband polarization-entangled photon pairs distributed over a WDM link for qubit networks. AB - We present a bright, narrowband, portable, quasi-phase-matched two-crystal source generating polarization-entangled photon pairs at 809 nm and 1555 nm at a maximum rate of 1.2 x 10(6) s(-1) THz-1 mW(-1) after coupling to single-mode fiber. The quantum channel at 1555 nm and the synchronization signal gating the single photon detector are multiplexed in the same optical fiber of length 27 km by means of wavelength division multiplexers (WDM) having 100 GHz (0.8 nm) spacing between channels. This implementation makes quantum communication applications compatible with current high-speed optical networks. PMID- 19547009 TI - Radiation torque on nonspherical particles in the transition matrix formalism: erratum. AB - An erratum is presented to correct the errors in two equations in Sect. 3 of [Opt. Express 14, 9508--9521 (2006)]. PMID- 19547008 TI - Imaging of protein cluster sizes by means of confocal time-gated fluorescence anisotropy microscopy. AB - A time-resolved fluorescence anisotropy imaging method for studying nanoscale clustering of proteins or lipids was developed and evaluated. It is based on FRET between the identical fluorophores (homo-FRET), which results in a rapid depolarization of the fluorescence. The method employs the time-resolved fluorescence anisotropy decays recorded in a confocal microscope equipped with pulsed excitation and time-gated detection. From the decay the limiting anisotropy r(inf) was derived, which is a direct measure for the number of fluorophores per cluster. The method was evaluated by imaging GPI-GFP, a lipid raft marker. Small clusters were observed in the plasma membrane while the cytoplasm and the Golgi contained predominantly monomers. PMID- 19547010 TI - Experimental studies of far-field superlens for sub-diffractional optical imaging. AB - Contrary to the conventional near-field superlensing, subwavelength superlens imaging is experimentally demonstrated in the far-field. The key element is termed as a Far-field SuperLens (FSL) which consists of a conventional superlens and a nanoscale coupler. The evanescent fields from the object are enhanced and then converted into propagating fields by the FSL. By only measuring the propagating field in the far-field, the object image can be reconstructed with subwavelength resolution. As an example of this concept, we design and fabricate a silver structured one dimensional FSL. Experimental results show that feature resolution of better than 50nm is possible using current FSL design. PMID- 19547011 TI - Fully adaptive finite element based tomography using tetrahedral dual-meshing for fluorescence enhanced optical imaging in tissue. AB - We have developed fluorescence enhanced optical tomography based upon fully adaptive finite element method (FEM) using tetrahedral dual-meshing wherein one of the two meshes discretizes the forward variables and the other discretizes the unknown parameters to be estimated. We used the 8-subtetrahedron subdivision scheme to create the nested dualmesh in which each are independently refined. However, two tetrahedrons from the two different meshes pose an intersection problem that needs to be resolved in order to find the common regions that the forward variables (the fluorescent diffuse photon fluence fields) and the parameter estimates (the fluorescent absorption coefficients) can be mutually assigned. Using an efficient intersection algorithm in the nested tetrahedral environments previously developed by the authors, we demonstrate fully adaptive tomography using a posteriori error estimates. Performing the iterative reconstructions using the simulated boundary measurement data, we demonstrate that small fluorescent targets embedded in the breast simulating phantom in point illumination/detection geometry can be resolved at reasonable computational cost. PMID- 19547012 TI - 100 mW high efficient single pass SHG at 488 nm of a single broad area laser diode with external cavity using a PPLN waveguide crystal. AB - A frequency stabilized single broad area laser in a V-shaped external cavity is used for Second Harmonic Generation (SHG) in a waveguide channel with dimensions of 3 mum x 5 mum x 10 mm of a PPMgO: LN crystal. A maximum coupling efficiency of 63% could be obtained. An optical output power of 100.4 mW of visible light at 488 nm could be generated with 265 mW of coupled infrared light. This results in a single pass conversion efficiency of 37.8%. No photorefractive damage or saturation effects were observed. PMID- 19547013 TI - Near-infrared femtosecond laser crystallized poly-Si thin film transistors. AB - Polycrystalline silicon (poly-Si) thin film transistors (TFTs) fabricated by near infrared femtosecond laser annealing (FLA) are demonstrated. The FLA-annealed poly-Si channels exhibit low tail-state, deep-state, and midgap-state densities of grain traps. Characteristics such as field-effect mobility, threshold voltage, and subthreshold slope for FLA-annealed poly-TFTs are comparable to those of conventional approaches. A wide process window for annealing laser fluences was confirmed by examining the changes in electrical parameters for transistors with various channel dimensions. PMID- 19547014 TI - Measurement of the far field intensity distribution of a bent and cleaved fiber taper. AB - We have theoretically predicted and experimentally demonstrated mode conversion in fiber tapers subject to large adiabatic bending. The far field intensity distribution of the taper mode is imaged in part by cleaving the taper at the position of minimum diameter. PMID- 19547015 TI - Effective medium description of plasmonic metamaterials. AB - We demonstrate that the electromagnetic properties of a plasmonic metamaterial, composed of a perfectly conducting metal film perforated with an array of holes, can be effectively described by a structureless, three layer film. The enhanced transmission, first observed by Ebbessen, is identified with resonant tunneling in the equivalent three layer system and perfect transmission is shown to be possible below the critical thickness of a metamaterial. The nature of modes mediating perfect transmission is clarified. PMID- 19547016 TI - Polarization conversion in a silica microsphere. AB - We experimentally demonstrate controlled polarization-selective phenomena in a whispering gallery mode resonator. We observed efficient ( approximately 75%) polarization conversion of light in a silica microsphere coupled to a tapered optical fiber with proper optimization of the polarization of the propagating light. A simple model treating the microsphere as a ring resonator provides a good fit to the observed behavior. PMID- 19547017 TI - Radiation torque on nonspherical particles in the transition matrix formalism: erratum. AB - An erratum is presented to correct the errors in two equations in Sect. 3 of [Opt. Express 14, 9508--9521 (2006)]. PMID- 19547018 TI - Negative nonlinear refraction obtained with ultrashort laser pulses. AB - We present nonlinear refraction results for liquids methanol and acetic acid obtained with the Z-scan technique and 28 femtosecond (fs) 800 nm laser pulses. In contrast to the positive lensing effect obtained previously with picosecond and nanosecond laser pulses, a negative lensing effect is observed. The associated mechanism features the third-order polarization arising from the nonlinear response of the molecular skeletal motion that is driven into resonance through its electrostatic coupling to the valence electron cloud distorted by the fs laser field. PMID- 19547019 TI - Spectral variability of the particulate backscattering ratio. AB - The spectral dependency of the particulate backscattering ratio is relevant in the fields of ocean color inversion, light field modeling, and inferring particle properties from optical measurements. Aside from theoretical predictions for spherical, homogeneous particles, we have very limited knowledge of the actual in situ spectral variability of the particulate backscattering ratio. This work presents results from five research cruises that were conducted over a three-year period. Water column profiles of physical and optical properties were conducted across diverse aquatic environments that offered a wide range of particle populations. The main objective of this research was to examine the behavior of the spectral particulate backscattering ratio in situ, both in terms of its absolute magnitude and its variability across visible wavelengths, using over nine thousand 1-meter binned data points for each of five wavelengths of the spectral particulate backscattering ratio. Our analysis reveals no spectral dependence of the particulate backscattering ratio within our measurement certainty, and a geometric mean value of 0.013 for this dataset. This is lower than the commonly used value of 0.0183 from Petzold's integrated volume scattering data. Within the first optical depth of the water column, the mean particulate backscattering ratio was 0.010. PMID- 19547020 TI - Phase manipulated multi-beam holographic lithography for tunable optical lattices. AB - We present principle and technique to actively stabilize and control the phase differences between multi-laser beams that produce stable and adjustable intensity patterns. This technique is based on a novel optical set-up and on a closed loop control over the phase difference between each pair of the input laser fields. Tunable optical lattices are demonstrated by exciting FeTPPCl-doped liquid crystals with the variable intensity patterns. PMID- 19547021 TI - Continuous-wave, intracavity doubled, self-Raman laser operation in Nd:GdVO(4) at 586.5 nm. AB - We report a continuous-wave self-Raman laser based on diode-pumped Nd:GdVO(4) giving first-Stokes output at 1173 nm and intracavity frequency-doubled output at 586.5 nm. A maximum cw output power at 1173 nm of 2 W was obtained for diode pump powers of 22 W and a maximum cw power at 586 nm of 678 mW with 16.3 W pump power. Infrared and yellow powers were limited by thermal lensing in the gain medium and parasitic oscillations of weak Nd(3+) transitions. Quasi-cw operation at 50% duty cycle reduced the thermal load in the laser/Raman crystal, allowing cavity stability to be obtained near maximum available pump power (25.7 W) for which the maximum quasi-cw yellow output power was 1.88 W. PMID- 19547022 TI - An ion-implanted InP receiver for polarization resolved terahertz spectroscopy. AB - We report on the construction, optical alignment and performance of a receiver which is capable of recording the full polarization state of coherent terahertz radiation. The photoconductive detector was fabricated on InP which had been implanted with Fe(+) ions. The device operated successfully when it was gated with near infrared femtosecond pulses from either a Ti:sapphire laser oscillator or a 1 kHz regenerative laser amplifier. When illuminated with terahertz radiation from a typical photoconductive source, the optimized device had a signal to noise figure of 100:1 with a usable spectral bandwidth of up to 4 THz. The device was shown to be very sensitive to terahertz polarization, being able to resolve changes in polarization of 0.34 degrees. Additionally, we have demonstrated the usefulness of this device for (i) polarization sensitive terahertz spectroscopy, by measuring the birefringence of quartz and (ii) terahertz emission experiments, by measuring the polarization dependence of radiation generated by optical rectification in (110)-ZnTe. PMID- 19547023 TI - Polarization insensitive low-loss coupling technique between SOI waveguides and high mode field diameter single-mode fibers. AB - A polarization insensitive technique for highly efficient coupling between SOI waveguides and high mode field diameter single-mode fibers is reported. The proposed coupling structure is based on an inverted taper structure coupled to a fiber-adapted waveguide. The fiber-adapted waveguide is made by using the SiO(2) layer under the Si waveguiding layer of the SOI wafer thus avoiding the use of extra materials such as polymers. The proposed coupling structure is aimed for being integrated with V-groove auto-alignment techniques. Coupling losses of 3.5dB and 3.7dB to 8mum mode field diameter single-mode fibers have been estimated by means of 3D-BPM simulations for TE and TM polarizations respectively and a 1550nm input signal wavelength. PMID- 19547024 TI - Novel fluorescent probe for low density lipoprotein, based on the enhancement of Europium emission band. AB - We report here the observation of the enhancement of Europium-tetracycline complex emission in Low Density Lipoprotein (LDL) solutions. Europium emission band of tetracycline solution containing Europium (III) chloride hexahydrate was tested to obtain effective enhancement in the presence of native LDL and oxidized LDL. Europium emission lifetime in the presence of lipoproteins was measured, resulting in a simple method to measure the lipoproteins quantity in an aqueous solution at physiological pH. This method shows that the complex can be used as a sensor to determine the different states of native and oxidized LDL in biological fluids. PMID- 19547026 TI - Highly efficient fluorescence of a fluorescing nanoparticle with a silver shell. AB - Spontaneous emission (SE) rate and the fluorescence efficiency of a bare fluorescing nanoparticle and the nanoparticle with a silver nanoshell are analyzed rigorously by using a classical electromagnetic approach with the consideration of the nonlocal effect of the silver nano-shell. The dependences of the SE rate and the fluorescence efficiency on the core-shell structure are carefully studied and the physical interpretations of the results are addressed. The results show that the SE rate of a bare nanoparticle is much slower than that in the infinite medium by almost an order of magnitude and consequently the fluorescence efficiency is usually low. However, by encapsulating the nanoparticle with a silver shell, highly efficient fluorescence can be achieved as a result of a large Purcell enhancement and high out-coupling efficiency for a well-designed core-shell structure. We also show that a higher SE rate may not offer a larger fluorescence efficiency since the fluorescence efficiency not only depends on the internal quantum yield but also the out-coupling efficiency. PMID- 19547025 TI - Broadly tunable high-power Yb:Lu(2)O(3) thin disk laser with 80% slope efficiency. AB - We report on efficient laser operation of high quality crystalline Yb(3+):Lu(2)O(3) in thin disk configuration. Using doping concentrations between 1 at.% to 3 at.% and disk thicknesses between 0.08mm and 0.45mm the optimum crystal parameters have been determined. Pumped at 976 nm the laser operates at 1034 nm and 1080 nm. With a 0.25mm thick 3 at.% Yb:Lu(2)O(3) disk 32.6W of output power at 45.3W incident pump power with a slope efficiency of 80% and a resulting optical-to-optical efficiency of 72% have been realized. These are the highest values in terms of slope efficiency as well as optical-to-optical efficiency for an Yb-doped thin disk laser reported so far. Using an 1mm birefringent filter continuous tuning from 987 nm to 1127 nm with more than 10Wof output power over a tuning range of 90 nm has been achieved. PMID- 19547027 TI - Tuning the far-field superlens: from UV to visible. AB - A far-field optical superlens, which is able to form sub-diffraction- limited images in the far field at UV wavelength, was recently demonstrated. In current work we present two methods to tune the working wavelength from UV to visible by tuning either the permittivity of the surrounding medium or that of the metal. A practical design is provided for each method. The tunable far-field superlens enables possible applications of the far-field superlens in sub-diffraction limited imaging and sensing over a wide range of wavelength. PMID- 19547028 TI - High-speed three-dimensional human retinal imaging by line-field spectral domain optical coherence tomography. AB - Line-field spectral domain optical coherence tomography (LF-SDOCT) has been developed for very high-speed three-dimensional (3D) retinal imaging. By this technique, the A-line rate significantly improved to 823,200 A-lines/s for single frame imaging and 51,500 A-lines/s for continues frame imaging. The frame rate at continues frame imaging is 201 fps. This 3D acquisition speed is more than two fold higher acquisition speed than the standard flying spot SD-OCT. In this paper, the integration time of the camera was optimized for the in vivo retinal measurement and the degradation of the lateral resolution due to the ocular aberrations was suppressed by introducing the pupil stop. Owing to an optimal integration time, the motion artifact can be significantly suppressed. Also a pupil stop was employed in order to enhance the contrast of the OCT image for the effect of ocular aberrations. The in vivo 3D retinal imaging with 256 cross sectional images (256 A-lines/image) was successfully performed in 1.3 seconds, corresponding to 0.8 volume/s. The maximum on-axis system sensitivity was measured to be 89.4 dB at a depth of 112 mum with an axial resolution of 7.4 mum in tissue. It is shown that LF-SDOCT might have a sensitivity advantage in comparison to the flying spot SD-OCT in the ultra high-speed acquisition mode. PMID- 19547029 TI - Large depth-high resolution full 3D imaging of the anterior segments of the eye using high speed optical frequency domain imaging. AB - Three dimensional rapid large depth range imaging of the anterior segments of the human eye by an optical frequency domain imaging system is presented. The tunable source spans from 1217 to 1356 nm with an average output power of 60 mW providing a measured axial resolution of 10 mum in air based on the coherence envelope. The effective depth range is 4 mm, defined as the distance over which the sensitivity drops by 6 dB, achieved by frequency shifting the optical signal using acousto optic modulators. The measured maximum sensitivity is 109 dB at a sample arm power of 14.7mW and A-lines rate of 43,900 per second. Images consisting of 512 depth profiles are acquired at an acquisition rate of 85 frames per second. We demonstrate an optical frequency domain imaging system capable of mapping in vivo the entire area of the human anterior segment (13.4 x 12 x 4.2 mm) in 1.4 seconds. PMID- 19547030 TI - Delivery of sub-100fs pulses through 8m of hollow-core fiber using soliton compression. AB - We report soliton compression in a tapered hollow-core photonic bandgap fiber. We compress unchirped 195fs input pulses at 800 nm wavelength to less than 100fs after single-mode propagation through 8m of fiber, at pulse energies of around 50nJ. PMID- 19547031 TI - Plasmonic optical properties of a single gold nano-rod. AB - Polarization-contrast microscopy coupled with an atomic force microscope is utilized to attain far-field optical images of the multipolar surface plasmon resonance (SPR) modes of single gold nano-rod. Modulated standing modes resulted from the interference of longitudinal SPR modes and incident light are observed and studied. By counting the average distance of adjacent beats on this single gold nano-rod, the wave vector of longitudinal SPR modes can be obtained. We found a linear relationship between the wave vectors of the incident light and the induced SPR modes. Experimental results demonstrate a feasible way on acquiring plasmonic optical properties from an individual single gold nano-rod. PMID- 19547032 TI - Dielectrically actuated liquid lens. AB - A packaged liquid lens driven by the dielectric force was demonstrated. The liquid lens consisted of a low dielectric constant droplet and a high dielectric constant sealing liquid. The two non-conductive liquids were sealed in a chamber under the condition of iso-density. Focal length of a liquid lens with an aperture of 3mm changed from 34mm to 12mm in the range of 0-200V. Hysteresis was observed in the liquid lens, with a maximum value measured of 12.5 degrees at 120 volts in terms of droplet's contact angle. The focal spot size measured approximately 80mum. Rise and fall times were 650ms and 300ms, respectively. The lens consumed 1mW of power when applying a 200 volt, 1 kHz signal. The longitudinal and transverse spherical aberrations were estimated to be nearly invariant when the focal length exceeded 20mm. PMID- 19547033 TI - Increased cross-correlation in cascaded four-wave mixing processes. AB - We report the measurement of increased noise cross-correlation between stokes and anti-stokes beams created in cascaded four-wave mixing processes with dual pump wavelengths. This method may be useful in creating highly correlated twin beams for various applications including quantum information processing. PMID- 19547034 TI - Empirical algorithms for ocean optics parameters. AB - As part of the Worldwide Ocean Optics Database (WOOD) Project, The Johns Hopkins University Applied Physics Laboratory has developed and evaluated a variety of empirical models that can predict ocean optical properties, such as profiles of the beam attenuation coefficient computed from profiles of the diffuse attenuation coefficient. In this paper, we briefly summarize published empirical optical algorithms and assess their accuracy for estimating derived profiles. We also provide new algorithms and discuss their applicability for deriving optical profiles based on data collected from a variety of locations, including the Yellow Sea, the Sea of Japan, and the North Atlantic Ocean. We show that the scattering coefficient (b) can be computed from the beam attenuation coefficient (c) to about 10% accuracy. The availability of such relatively accurate predictions is important in the many situations where the set of data is incomplete. PMID- 19547035 TI - Optical testing using the transport-of-intensity equation. AB - The transport-of-intensity equation links the intensity and phase of an optical source to the longitudinal variation of its intensity in the presence of Fresnel diffraction. This equation can be used to provide a simple, accurate spatial phase measurement for optical testing of flat surfaces. The properties of this approach are derived. The experimental demonstration is performed by quantifying the surface variations induced by the magnetorheological finishing process on laser rods. PMID- 19547036 TI - Colored object recognition by digital holography and a hydrogen Raman shifter. AB - Multi-wavelength holography is demonstrated with a H(2) Raman shifter that is pumped with an elliptically-polarized pulsed 532 nm beam to produce temporally coherent, intense, polarized output lines. Digital holograms of two-dimensional colored objects are recorded using Raman output lines at 630.4 nm (S(05), Red), 532 nm (Rayleigh, Green) and 435.7 nm (aS(10), Blue). Object reconstruction is done numerically via the convolution method and colored object recognition is achieved by multi-channel correlation of the Red, Green, and Blue reconstructions of the reference and the target object. PMID- 19547037 TI - Generalized dressed and doubly-dressed multi-wave mixing. AB - We present a theoretical treatment for generalized dressed and doubly-dressed multi-wave mixing processes. Co-existing four-wave mixing (FWM), six-wave mixing (SWM) and eight-wave mixing processes have been considered in a closed-cycle five level system. Due to constructive interference of the secondarily-dressed and primarily-dressed excitation pathways, the FWM and SWM signals are greatly enhanced. The dually enhanced FWM channels are opened simultaneously. PMID- 19547038 TI - Photoluminescence enhancement in nanoimprinted photonic crystals and coupled surface plasmons. AB - A method to enhance the photoluminescence of dye chromophores-loaded by coupling the emission to surface plasmons in nanoimprinted photonic crystals is reported. A 9-fold enhancement in the spontaneous emission intensity of a rhodamine-doped polymer film is achieved on a silver layer due to surface plasmon excitation. By changing the surface plasmon frequency, this enhancement can be suppressed. When the polymer film is patterned by nanoimprint lithography with a twodimensional photonic crystal the photoluminescence intensity increases up to 27 times compared to unpatterned samples on a quartz substrate. PMID- 19547039 TI - Ultrafast all-optical Nth-order differentiator based on chirped fiber Bragg gratings. AB - In this letter we present a technique for the implementation of Nth-order ultrafast temporal differentiators. This technique is based on two oppositely chirped fiber Bragg gratings in which the grating profile maps the spectral response of the Nth-order differentiator. Examples of 1(st), 2(nd), and 4(th) order differentiators are designed and numerically simulated. PMID- 19547040 TI - Extended depth of focus imaging with birefringent plate. AB - In this paper we present a simple approach to obtain extended depth of field for any optical imaging system just by adding a birefringent plate between the lens and the detector. The width of the plate is properly designed such that one polarization state contains in-focus near field information while the other polarization state contains in-focus far field details. Both images are superimposed one on top of the other and thus an all-optical spatially sharp imaging is obtained containing both fields. The width of the plate is also designed such that there is a longitudinal overlapping of the two regions (the near and the far) such that continuously well focused imaging is generated. The presented approach for extending the depth of focus is significantly simple compared to the use of birefringent and bi-focal lenses published recently. Preliminary numerical as well as experimental results verify the proposed approach. PMID- 19547041 TI - The Gouy phase shift in nonlinear interactions of waves. AB - We theoretically analyze the influence of the Gouy phase shift on the nonlinear interaction between waves of different frequencies. We focus on chi((2))interaction of optical fields, e.g. through birefringent crystals, and show that focussing, stronger than suggested by the Boyd-Kleinman factor, can further improve nonlinear processes. An increased value of 3.32 for the optimal focussing parameter for a single pass process is found. The new value builds on the compensation of the Gouy phase shift by a spatially varying, instead constant, wave vector phase mismatch. We analyze the single-ended, singly resonant standing wave nonlinear cavity and show that in this case the Gouy phase shift leads to an additional phase during backreflection. Our numerical simulations may explain ill-understood experimental observations in such devices. PMID- 19547042 TI - All-optical animation projection system with rotating fieldstone. AB - A simple and compact rewritable holographic memory system using a fieldstone of Ulexite is proposed. The role of the fieldstone is to impose random patterns on the reference beam to record plural images with the random-reference multiplexing scheme. The operations for writing and reading holograms are carried out by simply rotating the fieldstone in one direction. One of the features of this approach is found in a way to generate random patterns without computer drawings. The experimental study confirms that our system enables the smooth readout of the stored images one after another so that the series of reproduced images are projected as an animation. PMID- 19547043 TI - Optical measurement of rates of dissipation of temperature variance due to oceanic turbulence. AB - Inhomogeneities in the refractive index induced by temperature fluctuations in turbulent flows have the effect of scattering light in near-forward angles. We have developed a method that extracts the rate of Temperature Variance Dissipation (TVD) and its spectrum from the properties of light scattering and have built an instrument - Optical Turbulence Sensor (OTS) - that implements the method. OTS uses a linear wavefront sensing Hartmann array and allows for nearly instantaneous measurements of temperature variance in turbulent flows. The instrument has been tested in an situ experiment carried out from a drifting vessel at a site off the coast of Newport, Oregon. Here we compare the temperature variance measured by OTS and its spectra with both theoretical predictions and with spectra obtained from a fast thermistor sensor. PMID- 19547044 TI - Real-time dual-wavelength digital holographic microscopy with a single hologram acquisition. AB - A technique to perform two-wavelengths digital holographic microscopy (DHM) measurements with a single hologram acquisition is presented. The vertical measurement range without phase ambiguity is extended to the micron-range, thanks to the resulting synthetic wavelength defined by the beating of two wavelengths with a separation of about 80nm. Real-time dual-wavelength imaging is made possible by using two reference waves having different wavelengths and propagation directions for the hologram recording. The principle of the method is exposed and experimental results concerning a 1.2mum m high test sample as well as a moving micro-mirror are presented. To the extent of our knowledge, this is the first time that real-time synthetic beat-wavelength digital holography measurements are reported. PMID- 19547045 TI - Experiment on PMMA models to predict the impact of corneal refractive surgery on corneal shape: Comment. AB - Recently, two papers "Dorronsoro et al., Experiment on PMMA models to predict the impact of corneal refractive surgery on corneal shape, Opt. Express 14, 6142 (2006)" and "JR Jimenez et al., Deviations of Lambert-Beer's law affect corneal refractive parameters after refractive surgery, Opt. Express 14, 5411 (2006)" have been published on an important question in corneal refractive surgery: to explain the differences between post-surgical corneal elevation maps and those predicted after theoretical ablation. An analysis of Dorronsoro et al. data demonstrates that the failures in Lambert-Beer's law could be in the origin of shape discrepancies. New models and experimental data on deviations of Lambert Beer's law might help to minimize post-surgical corneal discrepancies and thus optimise eye emmetropization. PMID- 19547046 TI - Experiments on PMMA model to predict the impact of corneal refractive surgery on corneal shape: Reply. AB - A reply to the comment by Jimenez et al. directed to the paper of "Experiments on PMMA model to predict the impact of corneal refractive surgery on corneal shape" by Dorronsoro et al., Opt. Express, 14, 6142-6156 (2006). PMID- 19547047 TI - 1310-nm quantum key distribution system with up-conversion pump wavelength at 1550 nm. AB - We show that the performance of a 1310-nm quantum key distribution (QKD) system with up-conversion detectors pumped at 1550 nm is comparable with or better than that of current 1550-nm QKD systems with a pump at shorter wavelength. The nonlinearly-induced dark counts are reduced when the wavelength of the pump light is longer than that of the quantum signal. We have developed a 1550-nm pump up conversion detector for a 1310-nm QKD system, and we experimentally study the polarization sensitivity, pump-signal format, and various influences on the dark count rate. Using this detector in a proof-of-principle experiment, we have achieved a secure key rate of 500 kbit/s at 10 km and 9.1 kbit/s at 50 km in a 625-MHz, B92, polarization-coding QKD system, and we expect that the system performance could be improved further. PMID- 19547048 TI - A scaling law of photoionization in few-cycle regime. AB - In this paper, a scaling law of photoionization of atoms irradiated by intense, few-cycle laser pulses is established. The scaling law sets a relation to the phase-dependent ionization with the kinetic energy of photoelectrons, the duration and peak intensity of short pulses, and the ionization potential of the target atoms. We find that it will be advantageous to manifest the phase dependent photoionization by choosing the target atoms with larger ionization potential, using laser with smaller carrier-frequency, and increasing the pulse intensity. PMID- 19547049 TI - A hybrid CO(2) laser processing for silicon etching. AB - A novel approach to silicon (Si) etching has been demonstrated using glass assisted CO(2) laser processing. Conventional Si etching can be performed by wet etching, dry etching, Nd:YAG or UV lasers. No CO(2) laser was used to etch Si due to the absorption problem. We have etched Si with the assistance of glass beneath the Si. This approach changes light absorption behavior of Si and makes Si be etched from the top surface toward the interface. The new mechanism was discussed in viewpoint of the variation of electronic band structure, surface oxidation and light absorption of Si at high temperature. PMID- 19547050 TI - An optical threshold function based on polarization rotation in a single semiconductor optical amplifier. AB - Optical threshold functions are a basic building block for all-optical signal processing, and this paper investigates a threshold function design reliant on a single active element. An optical threshold function based on nonlinear polarization rotation in a single semiconductor optical amplifier is proposed. It functions due to an induced modification of the birefringence of a semiconductor optical amplifier caused by an externally injected optical control signal. It is shown that switching from both the TE to the TM mode and vice versa is possible. The measured results are supported by simulation results based on the SOA rate equations. PMID- 19547051 TI - Lasing characteristics of InAs quantum dot microcavity lasers as a function of temperature and wavelength. AB - A Strong temperature dependence of microdisk lasers and photonic crystal nanocavity lasers with InAs quantum dot active regions is reported. These lasers operate at 1.3 microm at room temperature under optical pumping conditions. T(0, microdisk) = 31 K. T(0, photonic crystal nanocavity) = 14 K. The lasing threshold dependence on the lasing wavelength is also reported. We observe a minimum absorbed threshold pump power of 9 microW. This temperature and wavelength dependent lasing behavior is explained qualitatively by a simple model which attributes the experimental observations predominantly to surface recombination at threshold and the high quality factors of these cavities. PMID- 19547052 TI - Microphotonic control of single molecule fluorescence correlation spectroscopy using planar optofluidics. AB - We demonstrate the implementation of fluorescence correlation spectroscopy (FCS) on a chip. Full planar integration is achieved by lithographic definition of sub picoliter excitation volumes using intersecting solid and liquid-core optical waveguides. Concentration dependent measurements on dye molecules with single molecule resolution are demonstrated. Theoretical modeling of the FCS autocorrelation function in microstructured geometries shows that the FCS behavior can be controlled over a wide range by tailoring the micro-photonic environment. The ability to perform correlation spectroscopy using silicon photonics without the need for free-space microscopy permits implementation of numerous diagnostic applications on compact planar optofluidic devices. PMID- 19547053 TI - Second harmonic generation imaging of fascia within thick tissue block. AB - Comparing the SHG image formation for thin sections of tail tendon fascia and skeletal muscle fascia, we demonstrate that the forward (F) and backward (B) SHG images are vastly different. In addition, despite the different arrangement of the collagen Type I fibrillar architecture forming these two fascias, their ratios of forward over backward signal (F/B) are nearly equal. SHG images of thick tissue blocks of the fascia-muscle unit show only backward features, as opposed to SHG images of tissue blocks of the fascia-tendon unit. These images are an amalgamation of forward and backward features due to the backscattering of forward components within tendon. These forward features disappear when this tissue block is immersed in glycerol as backscattering is hereby suppressed. PMID- 19547054 TI - Chi((2)) and Chi((3)) harmonic generation at a critical power in inhomogeneous doubly resonant cavities. AB - We derive general conditions for 100% frequency conversion in any doubly resonant nonlinear cavity, for both second- and third-harmonic generation via Chi?((2))and Chi((3)) nonlinearities. We find that conversion efficiency is optimized for a certain "critical" power depending on the cavity parameters, and assuming reasonable parameters we predict 100% conversion using milliwatts of power or less. These results follow from a semi-analytical coupled-mode theory framework which is generalized from previous work to include both Chi((2))and Chi((3))?media as well as inhomogeneous (fully vectorial) cavities, analyzed in the high-efficiency limit where down-conversion processes lead to a maximum efficiency at the critical power, and which is verified by direct finite difference time-domain (FDTD) simulations of the nonlinear Maxwell equations. Explicit formulas for the nonlinear coupling coefficients are derived in terms of the linear cavity eigenmodes, which can be used to design and evaluate cavities in arbitrary geometries. PMID- 19547055 TI - Spectral phase encoding of ultra-short optical pulse in time domain for OCDMA application. AB - We propose a novel reconfigurable time domain spectral phase encoding (SPE) scheme for coherent optical code-division-multiple-access application. In the proposed scheme, the ultra-short optical pulse is stretched by dispersive device and the SPE is done in time domain using high speed phase modulator. The time domain SPE scheme is robust to wavelength drift of the light source and is very flexible and compatible with the fiber optical system. Proof-of-principle experiments of encoding with 16-chip, 20 GHz/chip binary-phase-shift-keying codes and 1.25 Gbps data transmission have been successfully demonstrated together with an arrayed-wave-guide decoder. PMID- 19547056 TI - Single integrated device for optical CDMA code processing in dual-code environment. AB - We report on the design, fabrication and performance of a matching integrated optical CDMA encoder-decoder pair based on holographic Bragg reflector technology. Simultaneous encoding/decoding operation of two multiple wavelength hopping time-spreading codes was successfully demonstrated and shown to support two error-free OCDMA links at OC-24. A double-pass scheme was employed in the devices to enable the use of longer code length. PMID- 19547058 TI - Insight to UV-induced formation of laser damage on LiB(3)O(5) optical surfaces during long-term sum-frequency generation. AB - Microscopic investigations of UV-induced formation of laser damage on LiB(3)O(5) optical surfaces during long-term sum-frequency generation (SFG) uncovers a significant growth of a SiO(2)-amorphous layer spatially limited to the illuminated area. The layer gives rise to a catastrophic break-down of the LiB(3)O(5)-output surface upon long-term laser operation even at intensities far below the laser-induced damage threshold. The interaction of UV laser light, LiB(3)O(5) surface and foreign atoms in the ambient atmosphere is discussed in the frame of a two-step process for surface-damage formation. PMID- 19547057 TI - Feasibility of near-infrared diffuse optical spectroscopy on patients undergoing imageguided core-needle biopsy. AB - We describe a side-firing fiber optic sensor based on near-infrared spectroscopy for guiding core needle biopsy diagnosis of breast cancer. The sensor is composed of three side firing optical fibers (two source fibers and one detection fiber), providing two source-detector separations. The entire assembly is inserted into a core biopsy needle, allowing for sampling to occur at the biopsy site. A multi wavelength frequency-domain near-infrared instrument is used to collect diffuse reflectance in the breast tissue through an aperture on the biopsy needle before the tissue is removed for histology. Preliminary in vivo measurements performed on 10 normal or benign breast tissues from 5 women undergoing stereo- or ultrasound-guided core needle biopsy show the ability of the system to determine tissue optical properties and constituent concentrations, which are correlated with breast tissue composition derived from histopathology. PMID- 19547059 TI - QEPAS based detection of broadband absorbing molecules using a widely tunable, cw quantum cascade laser at 8.4 mum. AB - Detection of molecules with wide unresolved rotationa-lvibrational absorption bands is demonstrated by using Quartz Enhanced Photoacoustic Spectroscopy and an amplitude modulated, high power, thermoelectrically cooled quantum cascade laser operating at 8.4 mum in an external cavity configuration. The laser source exhibits single frequency tuning of 135 cm-1 with a maximum optical output power of 50 mW. For trace-gas detection of Freon 125 (pentafluoroethane) at 1208.62 cm 1 a normalized noise equivalent absorption coefficient of NNEA=2.64x10(-9) cm?( 1)W/Hz(1/2)was obtained. Noise equivalent sensitivity at ppbv level as well as spectroscopic chemical analysis of a mixture of two broadband absorbers (Freon 125 and acetone) with overlapping absorption spectra were demonstrated. PMID- 19547060 TI - Population study of the variation in monochromatic aberrations of the normal human eye over the central visual field. AB - We present data analysis for ocular aberrations of 60 normal eyes measured with a Hartmann-Shack (HS) wavefront sensor (WFS). Aberration measurements were made on axis and at 5 degree field angles in the nasal, inferior, temporal and superior semi-meridians. Particular attention is given to aberration distributions and possible strategies for aberration correction are discussed. A versatile HS WFS was designed and constructed with features of simultaneous pupil centre determination, off-axis capability, real-time data displays, and efficient lenslet sampling orientation. The subject alignment is achieved by the use of a parallel channel that is recombined with the sensing channel to simultaneously image the eye and the HS spots onto a single CCD. The pupil centre is determined using this image of the eye (iris edge), rather than the HS spots. The optical design includes a square lenslet array positioned with its diagonals aligned with the most typical principal astigmatic meridians of the eye. This favourable orientation helps to enlarge the dynamic range of the WFS. The telecentric re imaging of the HS spots increases the robustness of the system to defocus in the event of CCD misalignment. PMID- 19547061 TI - Optical tomography from focus. AB - A model and a method providing a 3D reconstruction of a given translucent object from a series of image acquisitions performed with various focus tunings is proposed. The object is imaged by transmission; refraction, reflection and diffusion effects are neglected. It is modeled as a stack of translucent parallel slices and the acquisition process can be described by a set of linear equations. We propose an efficient inversion technique with O(n) complexity, allowing practical applications with a simple laptop computer in a very reasonable time. Examples of results obtained with a simulated 3D translucent object are presented and discussed. PMID- 19547062 TI - Registration scheme suitable to Mueller matrix imaging for biomedical applications. AB - Most Mueller matrix imaging polarimeters implement sequential acquisition of at least 16 raw images of the same object with different incident and detected light polarizations. When this technique is implemented in vivo, the unavoidable motions of the subject may shift and distort the raw images to an extent such that the final Mueller images cannot be extracted. We describe a registration algorithm which solves this problem for the typical conditions of in vivo imaging, e.g. with spatially inhomogeneous medium to strong depolarization. The algorithm, based on the so called "optical flow," is validated experimentally by comparing the Mueller images of a pig skin sample taken in static and in dynamic conditions. PMID- 19547063 TI - Investigations on acousto-optically Q-switched frequency-shifted feedback laser with Nd:YVO(4) crystal. AB - An acousto-optically Q-switched frequency-shifted feedback (FSF) diode-pumped laser with a Nd:YVO(4) crystal is demonstrated. The laser emits more than 1W average power with multi-ten ns duration at up to 50 kHz repetition rate, and peak power higher than 1 kW is achieved at frequency of 20 kHz. This kind of pulsed laser has special value for some applications with outstanding features of broadband continuous spectrum and chirping frequency in the oscillation spectrum. Simulation results of the FSF laser through rate equations in Q-switched regime are also presented. PMID- 19547064 TI - All-optic scheme for automatic polarization division demultiplexing. AB - We describe a cost effective scheme to automatically separate two polarization channels in a polarization division multiplexing (PDM) system, without having to modify the existing transmitter or receiver electronics or software. We experimentally validate the concept by achieving an extinction ratio of more than 28-dB between two demultiplexed channels. Finally, we successfully demonstrate the PDM scheme in a 1.12-Tb/s (14x2x40-Gb/s) system over 62-km of transmission fiber. PMID- 19547065 TI - Generation of fractal speckles by means of a spatial light modulator. AB - It was shown in previous studies that, when a diffuser is illuminated by coherent light with an average spatial intensity distribution obeying a negative power function, the scattered field in the Fraunhofer diffraction region exhibits random fractal properties. The method employed so far for producing such fields has a disadvantage in that generated speckle intensities are low due to small transmittance of fractal apertures used in the illumination optics. To overcome this disadvantage, a generation of fractal speckles by means of a spatial light modulator is proposed. The principle is explained and experimental results are also shown. PMID- 19547067 TI - Optical properties of surface plasmon resonances of coupled metallic nanorods. AB - We present a systematic study of optical antenna arrays, in which the effects of coupling between the antennas, as well as of the antenna length, on the reflection spectra are investigated and compared. Such arrays can be fabricated on the facet of a fiber, and we propose a photonic device, a plasmonic optical antenna fiber probe, that can potentially be used for in-situ chemical and biological detection and surface-enhanced Raman scattering. PMID- 19547066 TI - Satellite remote sensing reveals regional tropospheric aerosol trends. AB - The Global Aerosol Climatology Project data product based on analyses of channel 1 and 2 AVHRR radiances shows significant regional changes in the retrieved optical thickness of tropospheric aerosols which had occurred between the volcano free periods 1988-91 and 2002-05. These trends appear to be generally plausible, are consistent with extensive sets of long-term ground-based observations throughout the world, and may increase the trustworthiness of the recently identified downward trend in the global tropospheric aerosol load. PMID- 19547068 TI - Femtosecond pulse synthesis by efficient second-harmonic generation in engineered quasi phase matching gratings. AB - We numerically design quasi-phase matched crystals with domains of arbitrary sizes for second harmonic generation by femtosecond pulses, taking into account both group velocity mismatch and dispersion. An efficient simulated-annealing algorithm is developed to design quasi-phase matching gratings which can yield the desired amplitude and phase of second-harmonic pulses in the presence of pump depletion. The method is illustrated with reference to single, double-hump and chirped fs Gaussian pulses in a lithium niobate crystal. PMID- 19547069 TI - Measurement of transient nonlinear refractive index in gases using xenon supercontinuum single-shot spectral interferometry. AB - A highly stable version of spectral interferometry is demonstrated, allowing single shot measurement of ultrafast high field processes using modest energy lasers, with pump and probe pulses totaling less than 1 mJ. The technique makes possible reconstruction of ultrafast refractive index transients with one dimensional spatial resolution, limited only by the bandwidth of the supercontinuum pulse (~100 nm) and instrument resolution. The ultrafast nonlinear Kerr effect in glass, and in Ar, N(2), and N(2)O gases is measured, along with plasma generation in Ar. The inertial contribution to the nonlinear index from N(2) and N(2)O molecular rotation is also observed. PMID- 19547070 TI - Liquid-crystal intraocular adaptive lens with wireless control. AB - We present a prototype of an adaptive intraocular lens based on a modal liquid crystal spatial phase modulator with wireless control. The modal corrector consists of a nematic liquid-crystal layer sandwiched between two glass substrates with transparent low- and high-ohmic electrodes, respectively. Adaptive correction of ocular aberrations is achieved by changing the amplitude and the frequency of the applied control voltage. The convex-shaped glass substrates provide the required initial focusing power of the lens. A loop antenna mounded on the rim of the lens delivers an amplitude-modulated radio frequency control signal to the integrated rectifier circuit that drives the liquid-crystal modal corrector. In vitro measurements of a 5-mm clear aperture prototype with an initial focusing power of +12.5 diopter, remotely driven by a radio-frequency control unit at ~6 MHz, were carried out using a Shack-Hartmann wavefront sensor. The lens based on a 40-mum thick liquid-crystal layer allows for an adjustable defocus of 4 waves, i. e. an accommodation of ~2.51 dioptres at a wavelength of 534 nm, and correction of spherical aberration coefficient ranging from -0.8 to 0.67 waves. Frequency-switching technique was employed to increase the response speed and eliminate transient overshoots in aberration coefficients. The full-scale settling time of the adaptive modal corrector was measured to be ~4 s. PMID- 19547071 TI - Spontaneous parametric down-conversion in periodically poled KTP waveguides and bulk crystals. AB - We present a theoretical and experimental comparison of spontaneous parametric down-conversion in periodically poled waveguides and bulk KTP crystals. We measured a waveguide pair generation rate of 2.9.10(6) pairs/s per mWof pump in a 1-nm band: more than 50 times higher than the bulk crystal generation rate. PMID- 19547072 TI - Multiple-channel silicon micro-resonator based filters for WDM applications. AB - We demonstrate predictable resonance wavelength shifts in silicon micro resonators by varying their perimeters using high-resolution lithography. The linear coefficient between the resonance wavelength shifts and the perimeter changes is determined with detailed experiments, and found to be nearly constant across the C and L bands in telecommunications. This empirical coefficient is also compared to that obtained from simulations on straight waveguides. Based on the linear model, without post-fabrication trimming or tuning, an eight-channel wavelength de-multiplexer with reasonably predicted average channel spacing ~ 1.8+/-0.1 nm (3dB bandwidth ~ 0.7+/-0.1 nm) is demonstrated at telecommunication bands in a silicon chip for the first time. This filter has out-of-band rejection ratio ~ 40 dB, low channel crosstalk 10(4). PMID- 19547117 TI - Electrically and mechanically induced long period gratings in liquid crystal photonic bandgap fibers. AB - We demonstrate electrically and mechanically induced long period gratings (LPGs) in a photonic crystal fiber (PCF) filled with a high-index liquid crystal. The presence of the liquid crystal changes the guiding properties of the fiber from an index guiding fiber to a photonic bandgap guiding fiber - a so called liquid crystal photonic bandgap (LCPBG) fiber. Both the strength and resonance wavelength of the gratings are highly tunable. By adjusting the amplitude of the applied electric field, the grating strength can be tuned and by changing the temperature, the resonance wavelength can be tuned as well. Numerical calculations of the higher order modes of the fiber cladding are presented, allowing the resonance wavelengths to be calculated. A high polarization dependent loss of the induced gratings is also observed. PMID- 19547118 TI - Surface waves and volume waves in a photonic crystal slab. AB - The volume and surface waves supported by a 2D PC slab with termination condition are systematically studied using the rigorous modematching method incorporating the Floquet's solutions. It is interesting to observe that the surface waves are caused by the perturbation of the PC-slab modes from the imposed termination condition, enabling the transition from volume wave to surface wave. The perturbed dispersion curves and electric field strength distribution over the structure are drawn together with the unperturbed ones (without termination condition) to identify the type of bound waves. PMID- 19547119 TI - Real-time actin-cytoskeleton depolymerization detection in a single cell using optical tweezers. AB - The cytoskeleton provides the backbone structure for the cellular organization, determining, in particular, the cellular mechanical properties. These are important factors in many biological processes, as, for instance, the metastatic process of malignant cells. In this paper, we demonstrate the possibility of monitoring the cytoskeleton structural transformations in optically trapped yeast cells (Saccharomyces cerevisiae) by tracking the forward scattered light via a quadrant photodiode. We distinguished normal cells from cells treated with latrunculin A, a drug which is known to induce the actin-cytoskeleton depolymerization. Since the proposed technique relies only on the inherent properties of the optical trap, without requiring external markers or biochemical sensitive spectroscopic techniques, it can be readily combined with existing optical tweezers setups. PMID- 19547120 TI - Generating Bragg solitons in a coherent medium. AB - In this paper we discuss the possibility of producing Bragg solitons in an electromagnetically induced transparency medium. We show that a coherent medium can be engineered to be a Bragg grating with a large Kerr nonlinearity through proper arrangements of light fields. The parameters of the medium can be easily controlled through adjusting the intensities and detunings of lasers. This scheme may provide an opportunity to study the dynamics of Bragg solitons with low power lights. PMID- 19547122 TI - Spectral domain polarization sensitive optical coherence tomography achieved by single camera detection. AB - We present a spectral domain polarization sensitive optical coherence tomography (PSOCT) system that incorporates: 1) a spectrometer configured with a single line scan camera for spectral interferogram detection, 2) a reference delay line assembly that provides a fixed optical pathlength delay between the lights of two orthogonal polarization states, and 3) a moving reference mirror that introduces a constant modulation frequency in the spatial spectral interferograms while the probe beam is scanned over the sample. The system utilizes the full range of complex Fourier plane for polarization sensitive imaging, where OCT images formed by the vertical and horizontal polarization beam components appear adjacent to each other. It is able to provide imaging of retardation, fast optic axis and backscattered intensity of the interrogated biological tissue. The system is experimentally demonstrated both in vitro and in vivo with an imaging rate at 10,000 A scans per second. PMID- 19547121 TI - High purity bright single photon source. AB - Using cavity-enhanced non-degenerate parametric down-conversion, we have built a frequency tunable source of heralded single photons with a narrow bandwidth of 8 MHz, making it compatible with atomic quantum memories. The photon state is 70% pure single photon as characterized by a tomographic measurement and reconstruction of the quantum state, revealing a clearly negative Wigner function. Furthermore, it has a spectral brightness of ~1,500 photons/s per MHz bandwidth, making it one of the brightest single photon sources available. We also investigate the correlation function of the down-converted fields using a combination of two very distinct detection methods; photon counting and homodyne measurement. PMID- 19547123 TI - The temperature sensitivity of Sagnac loop interferometer based on polarization maintaining side-hole fiber. AB - The temperature sensitivity of modal birefringence (dB/dT) of different types of polarization-maintaining fibers with side holes was measured using a Sagnac loop interferometer. The thermal expansion coefficient can be varied by controlling the amount of germanium doped in the core region. Using this method, dB/dT could be made higher (~10(-7)/ degrees C) than that of standard PMFs (~10(-8)/ degrees C) or comparable to that of standard PMFs (~10(-9)/ degrees C). PMID- 19547124 TI - Fully automated beam-alignment and single stroke guided manual alignment of counter-propagating multi-beam based optical micromanipulation systems. AB - In a previous paper [J. S. Dam et al, Opt. Express 15, 1923 (2007)] we demonstrated computerized "drag-and-drop" optical alignment of a counter propagating multi-beam based micromanipulation system. By inclusion of image analysis, we report here on the extension of this work to accommodate a completely automated beam-alignment process. Additionally, to maintain a cost effective and technically less demanding system architecture, we also report on a computer-guided manual alignment procedure. In the manual version, the computer analyzes the initial misalignment and the required compensations for each mirror in the system are calculated. Subsequently, the user is guided in adjusting the mirrors exactly by the requisite amount. This way, all mirrors only need to be moved once. The image analysis utilized in both calibration schemes employs a fitting algorithm to determine the position of beam-center with sub-pixel accuracy, thereby providing "better than human" alignment. PMID- 19547125 TI - High transmission recovery of slow light in a photonic crystal waveguide using a hetero groupvelocity waveguide. AB - High transmission of slow-light in a photonic crystal (PC) waveguide (WG) using a hetero group-velocity (Ht-V(g)) PC-WG was proposed and experimentally investigated. The Ht-V(g) WG, which comprises a low-group-velocity (L-V(g)) PC-WG section between two identical high-group- velocity (H-V(g)) PC-WGs, is designed to decrease the impedance mismatch of the L-V(g) PC-WG. The increase in transmittance of a propagating pulse was confirmed in the Ht-V(g) PC-WG even in the vicinity of the band-gap, whereas the homogeneous PC-WG showed a gradual decrease in transmittance with the pulse wavelength approaching the band-gap. The group index (n(g)) of the L-V(g) region in the Ht-V(g) PC-WG was measured by the cross-correlation method and attained a value above 20. On the other hand, the transmittance of the Ht-V(g) structure recovered approximately 16dB compared to the homogeneous L-V(g) WG having same n(g), 17. This recovery is mainly dominated by the coupling improvement due to the Ht-V(g) structure, around 12dB. These results indicate the effectiveness of the Ht-V(g) structure to use slow light in a PC-WG, which leads to various applications in PC-based optical devices. PMID- 19547126 TI - Interference and resonant cavity effects explain enhanced transmission through subwavelength apertures in thin metal films. AB - Transmission through an opaque Au film with a single subwavelength aperture centered in a smooth cavity between linear grating structures is studied experimentally and with a finite element model. The model is in good agreement with measured results and is used to investigate local field behavior. It shows that a surface plasmon polariton (SPP) is launched along the metal surface, while interference of the SPP with the incident light along with resonant cavity effects give rise to suppression and enhancement in transmission. Based on experimental and modeling results, peak location and structure of the enhancement/suppression bands are explained analytically, confirming the primary role of SPPs in enhanced transmission through small apertures in opaque metal films. PMID- 19547127 TI - High efficiency room temperature laser emission in heavily doped Yb:YLF. AB - We report the tunable, CW and quasi CW laser operation at room temperature of an highly doped (30% at.) Yb:YLF crystal longitudinally pumped by a fiber coupled laser diode array. The CW output power is 1.15 W vs. an absorbed pump power of 6 W, with a slope efficiency of 31%. In quasi-CW operation (20% duty factor @10 Hz) an output power of 4 W with an absorbed power of 9.5 W, and a slope efficiency of 62.8% were obtained. The tuning range spans from 1022 to 1075 nm. To our knowledge, these are among the best experimental results obtained at room temperature with Yb doped YLF. PMID- 19547128 TI - Negative refractions in two-dimensional photonic crystals formed by holographic lithography. AB - We present a study on negative refractions in the four lowest bands of two dimensional (2D) square lattices formed by holographic lithography (HL) and compare these features with those of a lattice of the same kind but with regular dielectric columns. The plane wave calculations and FDTD simulations have shown that in some bands or for some interfaces the negative refraction can only happen in holographic structures, and generally the rightness of holographic structures and regular structures of the same kind may be different. PMID- 19547129 TI - Analytical prediction of stable optical trapping in optical vortices created by three TE or TM plane waves. AB - A closed-form analytical expression of the force on an infinite lossless dielectric cylinder due to multiple plane wave incidences is proposed. The formula for a TE polarization is derived and completes our previous work which was limited to TM polarizations. A unified form of the analytical expression of the force is proposed and used to study the curvature of the one dimensional potential of an optical lattice created by the interference of three plane waves. It is shown that the points of zero curvature yield optical vortices which can be used to stably trap particles of particular sizes and index contrasts with the background. Under these circumstances, the trajectories of the particles can be assimilated to spirals whose centers correspond to the points of undetermined phase in the optical landscape. PMID- 19547130 TI - Nematicons across interfaces: anomalous refraction and reflection of solitons in liquid Crystals. AB - The robustness of nematicons, i. e. spatial solitons in nematic liquid crystals, can be exploited to implement counter-intuitive negative reflection and refraction schemes for optical signal manipulation at interfaces. PMID- 19547131 TI - Multi-view image fusion improves resolution in three-dimensional microscopy. AB - A non-blind, shift-invariant image processing technique that fuses multi-view three-dimensional image data sets into a single, high quality three-dimensional image is presented. It is effective for 1) improving the resolution and isotropy in images of transparent specimens, and 2) improving the uniformity of the image quality of partially opaque samples. This is demonstrated with fluorescent samples such as Drosophila melanogaster and Medaka embryos and pollen grains imaged by Selective Plane Illumination Microscopy (SPIM). The application of the algorithm to SPIM data yields high-resolution images of organ structure and gene expression, in some cases at a sub-cellular level, throughout specimens ranging from several microns up to a millimeter in size. PMID- 19547132 TI - Structural information within regularization matrices improves near infrared diffuse optical tomography. AB - Near-Infrared (NIR) tomographic image reconstruction is a non-linear, ill-posed and ill-conditioned problem, and so in this study, different ways of penalizing the objective function with structural information were investigated. A simple framework to incorporate structural priors is presented, using simple weight matrices that have either Laplacian or Helmholtz-type structures. Using both MRI derived breast geometry and phantom data, a systematic and quantitative comparison was performed with and without spatial priors. The Helmholtz-type structure can be seen as a more generalized approach for incorporating spatial priors into the reconstruction scheme. Moreover, parameter reduction (i.e. hard prior information) in the imaging field through the enforcement of spatially explicit regions may lead to erroneous results with imperfect spatial priors. PMID- 19547133 TI - Robust phase unwrapping by spinning iteration. AB - This work describes a rapid-phase unwrapping algorithm that combines the rapidity and simplicity of a path-dependent algorithm and the robustness of a path independent algorithm by rotating the phase map or the unwrapping direction 90 degrees after a scan in one direction. It offers a solution for noise contaminated phase data, which includes artifacts, complex-shaped borders, or regions of holes. The algorithm can be used in real-time processing. In addition, a phase-dislocation masking method is presented that can be used to detect and clean inconsistent data and improve the rms values of signal-to-noise ratio in unwrapped phase maps. PMID- 19547134 TI - Total internal reflection photonic crystal prism. AB - An integrated total internal reflection prism is demonstrated that generates a transversely localized evanescent wave along the boundary between a photonic crystal and an etched out trench. The reflection can be described by either the odd symmetry of the Bloch wave or a tangential momentum matching condition. In addition, the Bloch wave propagates through the photonic crystal in a negative refraction regime, which manages diffraction within the prism. A device with three input channels has been fabricated and tested that illuminates different regions of the reflection interface. The reflected wave is then sampled by a photonic wire array, where the individual channels are resolved. Heterodyne near field scanning optical microscopy is used to characterize the spatial phase variation of the evanescent wave and its decay constant. PMID- 19547135 TI - Simulation study on cascaded terahertz pulse generation in electro-optic crystals. AB - We studied cascaded optical rectification processes for intense terahertz (THz) pulse generation in electro-optic crystals using simulations based on one dimensional coupled propagation equations of THz and optical fields. We found that under ideal conditions of perfect phase matching and no absorption, cascaded optical rectification processes produce intense THz pulses with efficiencies exceeding the Manley-Rowe limit. Large red shifting of the pump light spectrum was observed. Effects of finite optical and THz absorption, phase mismatches, and pulse width were examined using parameters of a ZnTe crystal pumped by 800 nm pulses. THz field enhancement by multiple pulse pumping was also studied. PMID- 19547136 TI - Performance improvement of DPSK signal transmission by a phase-preserving amplitude limiter. AB - It is theoretically and experimentally shown that phase-preserving amplitude regeneration by an all-optical amplitude limiter using saturation of four-wave mixing in a nonlinear fiber can enhance DPSK transmission performance. The limiter suppresses amplitude fluctuations of the signal, by which the nonlinear phase noise caused by self-phase modulation of the transmission fiber is reduced. A 10-Gbit/s short-pulse DPSK transmission experiment shows that the limiter inserted either after a transmitter or inside a recirculating transmission loop can enhance the performance. Theoretical expressions for the linear and nonlinear phase noise are derived, with which the influence of imperfections of the limiter is examined. PMID- 19547137 TI - Combined effect of Raman and parametric gain on single-pump parametric amplifiers. AB - We investigate the combined effect of Raman and parametric gain on single-pump parametric amplifiers. The phasematched parametric gain is shown to depend strongly on the real part of the complex Raman susceptibility. In fused silica fibers this results in a significant reduction in the available parametric gain for signal detunings beyond 10 THz. We are able to experimentally measure this effect for signal detunings ranging from 7 to 22 THz. Finally we discuss the implications of these results for the design of broadband optical parametric amplifiers. PMID- 19547138 TI - Investigating nanoscale cellular dynamics with cross-sectional spectral domain phase microscopy. AB - We report on cross-sectional imaging of dynamic biological specimens using a spectral domain phase microscopy (SDPM) system capable of operating at a line rate of 19 kHz. This system combines the time-sensitive capabilities of SDPM with the multi-point acquisition features of related phase-sensitive techniques. The presented phase portraits and B-scan phase images of spontaneously beating embryonic cardiomyocytes and cytoplasmic flow in A. proteus offer insight into the nature and timing of the observed cellular phenomena, demonstrating the utility of this technique for dynamic cell studies. PMID- 19547139 TI - Polarized left-handed extraordinary optical transmission of subterahertz waves. AB - In this paper we design and measure a metamaterial polarizing device working in the sub-terahertz range. The polarizer is based on a modified version of our previous miniaturized Stacked Hole Array (SHA) structure, an arrangement that combines Extraordinary Optical Transmission (EOT) and Left-Handed Metamaterial (LHM) propagation even under Fresnel illumination. Here, we use a self complementary screen by connecting the holes of an EOT structure. Importantly, EOT remains and simultaneously total reflection is obtained for the orthogonal component. Moreover, by computing the dispersion diagram, we demonstrate that LHM propagation can be achieved for the principal polarization within the stop band of the orthogonal component, which propagates in other bands as a standard forward wave. Finally, we check our conjectures by measuring the transmission and reflection coefficients of screens milled on a low-loss microwave substrate. Measurements have been taken for 1 to 6 stacked wafers and they show clearly that the stack acts as a polarizer with left-handed characteristic. Our results open the way to design of novel polarization control metamaterials at Terahertz wavelengths. PMID- 19547140 TI - Detection of small particles in fluid flow using a self-mixing laser. AB - We describe a highly sensitive, real-time method of detecting small particles in a fluid flow by self-mixing laser Doppler measurement with a laser-diode-pumped, thin-slice solid-state laser with extremely high optical sensitivity. Asymmetric power spectra of the laser output modulated by the re-injected scattered light from the small particles moving in a dilute sample-flow, through a small-diameter glass pipe, were observed. The observed power spectra are shown to reflect the velocity distribution of the fluid flow, which obeys Poiseuille's law. Quick measurements of flow rate and kinetic viscosities of water-glycerol mixtures were also performed successfully. Measurable low-concentration limits for 262-nm polystyrene latex spheres and 3-mum red blood cells in a fluid flow were below 1 and 10 ppm, respectively, in the present self-mixing laser Doppler velocimeter system. PMID- 19547141 TI - Polarization and particle size dependence of radiative forces on small metallic particles in evanescent optical fields. Evidences for either repulsive or attractive gradient forces. AB - We have observed the motion of metallic particles above various optical waveguides injected by 1064nm radiation. Small gold particles (250nm diameter) are attracted towards the waveguide where the intensity of the optical field is maximum, and are propelled at high velocity (up to 350mum/s) along the waveguide due to radiation pressure. The behaviour of larger metallic particles (diameter >600nm) depends on the polarization of the evanescent field: for TM polarization they are attracted above the waveguide and propelled by the radiation pressure; for TE polarization they are expelled on the side of the waveguide and propelled at much smaller velocity. This is consistent with calculations of radiative forces on metallic particles by Nieto-Vesperinas et al. 3D-finite element method calculations carried out for our experimental situations confirm the observed dependence with the polarization of the field and the size of the particles. These observations open the way to the development of new microsystems for particles manipulations and sorting applications. PMID- 19547142 TI - Tuning the optical performance of surface quantum dots in InGaAs/GaAs hybrid structures. AB - GaAs spacer thicknesses are varied to tune the coupling between InGaAs surface quantum dots (QDs) and multilayers of buried QDs. Temperature and excitation intensity dependence of the photoluminescence together with time resolved photoluminescence reveal that coupling between layers of QDs and consequently the optical properties of both the surface and the buried QDs significantly depend on the GaAs spacer. This work provides an experimental method to tune and control the optical performance of surface QDs. PMID- 19547143 TI - Enhanced SPR sensitivity using periodic metallic structures. AB - A sinusoidal silver grating is used to create a six-fold enhancement of the SPR response compared to a flat surface. The grating parameters are chosen to create a surface plasmon bandgap and it is shown that the enhancement of the sensitivity to bulk sample index occurs when operating near the bandgap. The Kretschmann configuration is considered and the Boundary Element Method is used to generate the dispersion curves. PMID- 19547144 TI - Mode locking using stimulated Raman scattering. AB - Recent experiments have shown that the simple addition of a length of Raman shifting fibre in the cavity of a cw-pumped fibre laser can cause the laser to generate a stable train of pulses [Zhao and Jackson, Opt. Lett., 31 751 (2006)]. We show using a numerical model that this behavior is a new type of mode locking, driven by backward stimulated Raman scattering. This mode locking mechanism could also be applied to crystalline Raman laser systems to create a novel picosecond oscillator. PMID- 19547145 TI - Image based adaptive optics through optimisation of low spatial frequencies. AB - We present a wave front sensorless adaptive optics scheme for an incoherent imaging system. Aberration correction is performed through the optimisation of an image quality metric based upon the low spatial frequency content of the image. A sequence of images is acquired, each with a different aberration bias applied and the correction aberration is estimated from the information in this image sequence. It is shown, by representing aberrations as an expansion in Lukosz modes, that the effects of different modes can be separated. The optimisation of each mode becomes independent and can be performed as the maximization of a quadratic function, requiring only three image measurements per mode. This efficient correction scheme is demonstrated experimentally in an incoherent transmission microscope. We show that the sensitivity to different aberration magnitudes can be tuned by changing the range of spatial frequencies used in the metric.We also explain how the optimization scheme is related to other methods that use image sharpness metrics. PMID- 19547146 TI - Localized and delocalized modes in coupled optical micropillar cavities. AB - Optical micropillar Bragg cavities of different diameters and coupled by a small bridge have been realized experimentally by means of a focused ion beam system. The resonator modes in these coupled microcavities are either localized in one pillar or delocalized over the whole photonic structure, a fact that could be exploited to control the coupling between two spatially separated quantum dots, i.e. placed in different pillars, via the enhanced electromagnetic field in such a coupled microcavity. A simplified two dimensional simulation has been used to predict the resonant wavelengths and design the optical modes in these coupled Bragg cavities. PMID- 19547147 TI - Spectral filtering in a diode-pumped Nd:YLF regenerative amplifier using a volume Bragg grating. AB - Instrument-limited suppression of out-of-band Amplified Spontaneous Emission (ASE) is demonstrated in a Nd:YLF Diode-Pumped Regenerative Amplifier (DPRA) using a Volume Bragg Grating (VBG) as a spectrally selective reflective element. A VBG with 99.4% diffraction efficiency and 230-pm-FWHM reflection bandwidth produced a 43-pm- FWHM output spectral width in an unseeded DPRA compared to 150 pm FWHM in the same DPRA with no VBG. Instrument-limited ASE suppression is even observed when the DPRA seed pulse energy approaches the ASE background. PMID- 19547148 TI - Precise and long-term stabilization of the carrier-envelope phase of femtosecond laser pulses using an enhanced direct locking technique. AB - We demonstrate a long-term operation with reduced phase noise in the carrier envelope-phase (CEP) stabilization process by employing a double feedback loop and an improved signal detection in the direct locking technique [Opt. Express 13, 2969 (2005)]. A homodyne balanced detection method is employed for efficiently suppressing the dc noise in the f-2f beat signal, which is converted into the CEP noise in the direct locking loop working at around zero carrier envelope offset frequency (f(ceo)). In order to enhance the long-term stability, we have used the double feedback scheme that modulates both the oscillator pump power for a fast control and the intracavity-prism insertion depth for a slow and high-dynamic-range control. As a result, the in-loop phase jitter is reduced from 50 mrad of the previous result to 29 mrad, corresponding to 13 as in time scale, and the long-term stable operation is achieved for more than 12 hours. PMID- 19547149 TI - Tunable, Single-frequency, Diode-pumped 2.3mum VECSEL. AB - We report high-performance single-frequency operation of a directly diode-pumped GaSb-based vertical-external-cavity surface-emitting laser (VECSEL) at 2.3mum. Tunability of 70nm and a maximum single frequency output of 0.68W is demonstrated. PMID- 19547150 TI - Genetic optimization of photonic bandgap structures. AB - We investigate the use of a Genetic Algorithm (GA) to design a set of photonic crystals (PCs) in one and two dimensions. Our flexible design methodology allows us to optimize PC structures for specific objectives. In this paper, we report the results of several such GA-based PC optimizations. We show that the GA performs well even in very complex design spaces, and therefore has great potential as a robust design tool in a range of PC applications. PMID- 19547151 TI - Gas to crystal Effect on the Spectral Line Narrowing of MEH-PPV. AB - We report two emission bands corresponding to the spectral line narrowing (SLN) of the conjugated polymer [2-methoxy-5-(2'-ethylhexyloxy)-1, 4-phenylenevinylene] (MEH-PPV) in films. The SLN emission coming from the polymer chains closer to the glass substrate are at a different spectral position compared to the chains that lay further away from the glass substrate. We explain this phenomenon as a direct consequence of the "gas-to-crystal" effect. In solution form, as concentration was increased, and thus the proportion of aggregates, a decrease in the SLN bandwidth and a red shift of the emission peak was observed. PMID- 19547152 TI - High resolution optical time domain reflectometer based on 1.55mum up-conversion photon-counting module. AB - We implement a photon-counting Optical Time Domain Reflectometer (OTDR) at 1.55mum which exhibits a high 2-point resolution and a high accuracy. It is based on a low temporal-jitter photon-counting module at 1.55mum. This detector is composed of a periodically poled Lithium niobate (PPLN) waveguide, which provides a wavelength conversion from near infrared to visible light, and a low jitter silicon photon-counting detector. With this apparatus, we obtain centimetre resolution over a measurement range of tens of kilometres. PMID- 19547153 TI - Amplitude and phase reconstruction of photorefractive spatial bright-soliton in LiNbO3 during its dynamic formation by digital holography. AB - The time behaviour of bright spatial solitons in congruent undoped lithium niobate crystal is experimentally investigated. Full field characterization of the optical wavefront emerging from the crystal during the soliton formation process is performed by digital holographic method. Experimental results of the amplitude and phase maps of the field distribution at the exit face of the crystal allow the real-time monitoring of the evolution of the soliton beam from the application of the external field to the end of the process when the generation of the channel waveguide appears to be stable. The features of the dynamics of the soliton formation are visualized, analyzed and compared to a time dependent numerical model. PMID- 19547154 TI - On the study of pulse evolution in ultra-short pulse mode-locked fiber lasers by numerical simulations. AB - In this contribution we highlight several aspects concerning the numerical simulation of ultra-short pulse mode-locked fiber lasers by a non-distributed model. We show that for fixed system parameters multiple attractors are accessible by different initial conditions especially in the transient region between different mode-locking regimes. The reduction of multiple attractors stabilizing from different quantum noise fields to a single solution by gain ramping is demonstrated. Based on this analysis and model, different regimes of mode-locking obtained by varying the intra-cavity dispersion and saturation energy of the gain fiber are revised and it is shown that a regime producing linearly chirped parabolic pulses known from self-similar evolution is embedded in the wave-breaking free mode-locking regime. PMID- 19547155 TI - Tracking high amplitude auto-oscillations with digital Fresnel holograms. AB - Method for tracking vibrations with high amplitude of several hundreds of micrometers is presented. It is demonstrated that it is possible to reconstruct a synthetic high amplitude deformation of auto-oscillations encoded with digital Fresnel holograms. The setup is applied to the auto-oscillation of a clarinet reed in a synthetic mouth. Tracking of the vibration is performed by using the pressure signal delivered by the mouth. Experimental results show the four steps of the reed movement and especially emphasize the shocks of the reed on the mouthpiece. PMID- 19547156 TI - High-flux photon-pair source from electrically induced parametric down conversion after second-harmonic generation in single optical superlattice. AB - We present here a possible high-flux photon-pair source constructed by single lithium niobate optical superlattice (OSL) with a combined quasi-periodically and periodically poled structure, which is from the principle of electrically induced parametric down conversion (PDC) after second-harmonic generation (SHG), predicted by the united theory developed in this paper, in which SHG, PDC and electro-optic (EO) effect are comparably treated as two-order nonlinear effects. In the OSL, the e-polarized fundamental frequency photons are first converted to double frequency ones with the same polarization; then the PDC process is triggered by EO effect when the fundamental frequency photons are almost exhausted; finally, the double frequency photons are converted again to a series of two-photon pair of fundamental wave. It is demonstrated that at 100 degrees C, in a 20.2mm long OSL with a 30V / mm applied electric field, a 100MW/cm(2), 1080 nm laser beam can be translated to a flux of high-purity two-photon pairs with a conversion efficiency close to 100%; and for a longer OSL the pump intensity can be further lowered. The device can also act as an ultra-low field electro-optic switch. PMID- 19547157 TI - Left-handed material based on ferroelectric medium. AB - Left-handed metamaterials always gain the electromagnetic properties from the structure rather than inherit them directly from the materials they are composed of. In this article, a metamaterial was made using split-ring resonators and slabs of ferroelectric materials, where negative permittivity was realized by intrinsic properties of ferroelectric materials. Using a waveguide-based retrieval method, the permittivity and permeability of the metamaterials were experimentally retrieved, showing successfully the left-handed behaviors of the metamaterial over certain frequency band. PMID- 19547158 TI - Al/Ge co-doped large mode area fiber with high SBS threshold. AB - We propose a novel approach of making large effective area laser fiber with higher threshold for the stimulated Brillouin scattering (SBS) using Al/Ge co doping in the fiber core. The increased SBS threshold is achieved by reducing the acoustic-optic overlap integral while keeping the optical refractive index profile with a step structure. The manipulation of the overlap integral is done by adjusting the relative doping level between Al(2)O(3) and GeO(2) in the core. The mechanism is validated by detailed examples of numerical modeling. An Yb doped double clad fiber with the core co-doped with Al(2)O(3) and GeO(2) was fabricated by the OVD process. Measured acoustic velocity profile using a scanning acoustic microscope verified that the acoustic velocity in the fiber core changes with the design. An amplifier utilizing the fiber demonstrated that the proposed fiber yielded 6 dB higher SBS threshold than a fiber without using the co-doping scheme. PMID- 19547159 TI - Design of an ultracompact MMI wavelength demultiplexer in slot waveguide structures. AB - Based on a multimode interference (MMI) coupler in slot waveguide structures, an ultracompact wavelength demultiplexer operating at 1.30 and 1.55mum wavelengths is proposed and designed by using a full-vector mapped Galerkin mode solver and a modified three-dimensional full-vector beam propagation method. The tapered waveguide structures are applied to connect the input/output channels and the MMI section for reducing excess loss. The modal characteristics of the slot waveguides are analyzed and the evolution of the injected field in whole device are demonstrated. The results show that a MMI section of 119.8mum in length, which is only 27.5% length of that of the MMI coupler by using conventional rib waveguides, is achieved with the contrasts of 26.03 and 28.14dB at wavelengths 1.30 and 1.55mum, respectively, and the insertion losses are below 0.2dB at both wavelengths. PMID- 19547160 TI - Tuning the resonance frequency of Ag-coated dielectric tips. AB - A finite element model was built to investigate how to optimize localized plasmon resonances of an Ag-coated dielectric tip for tip-enhanced Raman spectroscopy (TERS). The relation between the resonance frequency, the electric field enhancement and the optical constant of the dielectric tip was numerically investigated. The results show that increasing the refractive index of the dielectric tip can significantly red shift the localized plasmon modes excited on the Ag-coated dielectric tip, and consequently alter the field enhancement. Moreover, the influence of the width of the resonance on the Raman enhancement was also considered. When taking all the factors into account, we find that an Ag coated low-refractive index dielectric tip provides the best Raman enhancement in the blue-green spectral range. This is consistent with our prior experimental results. PMID- 19547161 TI - Continuously-tunable, bit-rate variable OTDM using broadband SBS slow-light delay line. AB - We conceptually compare the advantages of the proposed slow-light-based tunable OTDM to conventional fiber-based fixed OTDM multiplexer. We experimentally demonstrate continuously-controllable OTDM of two 2.5-Gb/s return-to-zero (RZ) signals using broadband SBS-based slow-light as the tunable optical delay line. We show that the time slot of one signal path can be manipulated relative to the other by as much as 75-ps. This continuous slow light tunability dramatically enhances the OTDM system performance which results in a power penalty reduction of 9-dB for the multiplexed data stream. We also demonstrate variable-bit-rate OTDM by dynamically adjusting the tunable slow-light delay according to the input bit-rates. We show efficient two-by-one optical time multiplexing of three different input data streams at 2.5-Gb/s, 2.67-Gb/s and 5-Gb/s. PMID- 19547162 TI - Dispersion engineering of photonic crystal waveguides with ring-shaped holes. AB - The geometry of photonic crystal waveguides with ring-shaped holes is optimized to minimize dispersion in the slow light regime. We found geometries with a nearly constant group index in excess of 20 over a wavelength range of 8 nm. The origin of the low dispersion is related to the widening of the propagating mode close to the lower band gap edge. PMID- 19547163 TI - Surface enhanced ellipsometric contrast (SEEC) basic theory and lambda/4 multilayered solutions. AB - The fundamentals of a new high contrast technique for optical microscopy, named "Surface Enhanced Ellipsometric Contrast" (SEEC), are presented. The technique is based on the association of enhancing contrast surfaces as sample stages and microscope observation between cross polarizers. The surfaces are designed to become anti-reflecting when used in these conditions. They are defined by the simple equation r(p) + r(s) = 0 between their two Fresnel coefficients. Most often, this equation can be met by covering a solid surface with a single lambda/4 layer with a well defined refractive index. A higher flexibility is obtained with multilayer stacks. Solutions with an arbitrary number of all dielectric lambda/4 layers are derived. PMID- 19547164 TI - Calculation of effective permittivity, permeability, and surface impedance of negative-refraction photonic crystals. AB - We consider the eigen-fields of a two-dimensional negative-refraction photonic crystal and obtain negative effective permittivity and negative effective permeability. Effective permittivity, permeability, and surface impedance are calculated by averaging the eigen-fields. The value of the surface impedance is shown to be location-dependent and is validated by finite-difference time-domain simulations. The unique power propagation mechanism in the photonic crystal is demonstrated through time-evolution of eigen-fields. PMID- 19547165 TI - Heterodyne two beam Gaussian microscope interferometer. AB - We present a novel microscope interferometric technique based on the heterodinization of two Gaussian beams for measuring roughness of optical surfaces in microscopic areas. One of the beams is used as a probe beam, focussed and reflected by the surface under test. The second beam interferes with the first beam and introduces a time varying modulating signal. The modulating light beam is obtained from the first diffraction order of a Bragg cell. The two beams are superimposed and added coherently at the sensitive plane of a photodetector that integrates the overall intensity of the beams. We show analytically that it is possible to find appropriate working conditions in which the system has a linear response. Under these conditions, the size of the probe beam at the plane of detection as well as the amplitude of the time varying signal at the output of the photodetector, are both proportional to the local vertical height of the surface under test. As a narrow bandwidth amplifier is used to detect the time varying signal the system exhibits a high signal to noise ratio. We also include experimental results of the measurement of the topography of a sample consisting in a blazed-reflecting grating. PMID- 19547166 TI - A moderate-spectral-resolution transmittance model based on fitting the line-by line calculation. AB - A fast narrowband transmittance model, referred to as the Fast Fitting Transmittance Model (FFTM), is developed based on rigorous line-by- line (LBL) calculations. Specifically, monochromatic transmittances are first computed from a LBL model in a spectral region from 1 to 25000 cm(-1) for various pressures and temperatures ranging from 0.05 hPa to 1100 hPa and from 200 K to 320 K, respectively. Subsequently, the monochromatic transmittances are averaged over a spectral interval of 1 cm(-1) to obtain narrowband transmittances that are then fitted to various values of absorber amount. A database of fitting coefficients is then created that can be used to compute narrowband transmittances for an arbitrary atmospheric profile. To apply the FFTM to an inhomogeneous atmosphere, the Curtis-Godson (CG) approximation is employed to obtain the weighted effective coefficients. The present method is validated against the LBLRTM and also compared with the high-spectral-resolution measurements acquired by the Atmospheric Infrared Sounder (AIRS) and High-resolution Interferometer Sounder (HIS). With a spectral resolution of 1 cm(-1) and a wide spectral coverage, the FFTM offers a unique combination of numerical efficiency and considerable accuracy for computing moderate- to high-spectral-resolution transmittances involved in radiative transfer simulations and remote sensing applications. PMID- 19547167 TI - Tunable multi-wavelength SOA fiber laser based on a Sagnac loop mirror using an elliptical core side-hole fiber. AB - We propose and experimentally demonstrate a tunable multi-wavelength SOA fiber laser based on a Sagnac loop mirror using elliptical core side-hole fiber. We fabricated different types of elliptical core side-hole fiber with elliptical or circular cladding shape. The measured modal birefringence of the fabricated fiber was 1.7x10(-4) (elliptical shape) and 1.16x10(-4) (circular shape), respectively. By carefully adjusting the polarization controller inside the cavity, we could obtain 18 discrete channels with SNR over 30 dB and channel spacing of 0.8 nm and 10 discrete channels with SNR over 30 dB and channel spacing of ~1.4 nm. The proposed fiber laser was rather stable and the temporal power fluctuation was less than 0.8 dB. In addition, by thermally heating the elliptical core side-hole fiber in the Sagnac loop, we could obtain a tunable multi-wavelength fiber laser. PMID- 19547168 TI - Injection of ethanol into supercritical CO(2): Determination of mole fraction and phase state using linear Raman scattering. AB - For the pulsed injection of liquid ethanol into supercritical CO(2) inside an optically accessible chamber, for the first time to the best of our knowledge the spatially and temporally resolving linear Raman scattering technique was used to simultaneously determine the mole fraction and the corresponding phase state in the ethanol jet. The mole fraction was identified by calculating the ratio of the C-H band Raman signal (2950 cm(-1)) of ethanol and the CO(2) Raman signal. The magnitude of this ratio was found to be phase state sensitive. Thus, the phase state of the mixture of ethanol and CO(2) could be classified as being homogeneous liquid, homogeneous supercritical or not yet homogeneously mixed. PMID- 19547169 TI - An iterative phase retrieval algorithm for in-line x-ray phase imaging. AB - A general theoretical formulism for in-line phase x-ray imaging was presented with a corresponding linear formula in previous works. In this report, an iterative approach is introduced for phase retrieval with a nonlinear formula. The results of simulation showed that the iterative approach can retrieve the phase map more effectively with high efficiency and flexibility. PMID- 19547170 TI - Polarization and thickness dependent guiding in the photonic crystal slab waveguide. AB - We investigated the impact of the thickness of the twodimensional triangular photonic crystal (PC) on modal propagation along a PC slab waveguide. A single line defect optical waveguide in photonic crystal slab was designed by three dimensional finite difference time domain method, plane wave expansion and effective index methods. The thickness of the PC slab waveguide was optimized to provide modal propagation for both TE-like and TM-like polarizations within the normalized frequency band of a/lambda=0.26-0.268. PMID- 19547172 TI - Stress-induced focal splitting. AB - Under circularly polarized illumination, a trigonal birefringence pattern produces concentric rings of alternating right and left circular polarization. When a window with trigonal stress is placed in the pupil of an imaging system, the birefringence induces an axial splitting of the focus. We analyze and experimentally test this phenomenon and discuss applications to optical imaging. PMID- 19547171 TI - Towards a millivolt optical modulator with nano-slot waveguides. AB - We describe a class of modulator design involving slot waveguides and electro optic polymer claddings. Such geometries enable massive enhancement of index tuning when compared to more conventional geometries. We present a semi-analytic method of predicting the index tuning achievable for a given geometry and electro optic material. Based on these studies, as well as previous experimental results, we show designs for slot waveguide modulators that, when realized in a Mach Zehnder configuration, will allow for modulation voltages that are orders of magnitude lower than the state of the art. We also discuss experimental results for nano-slot waveguides. PMID- 19547173 TI - InP photonic wire waveguide using InAlAs oxide cladding layer. AB - We proposed a novel InP based photonic wire waveguide with an InAlAs oxide cladding. The InGaAsP/InAlAs-oxide structure in the vertical direction provides an ultrahigh index contrast waveguide, and it allows a bend radius of a few mum with no vertical leakage loss. The InP photonic wire waveguide with a 500x300-nm rectangular channel core (refractive index n ~ 3.36) and an InAlAs oxide cladding (n ~ 2.4) was numerically analyzed using the three-dimensional time-domain beam propagation method (3D TD-BPM). We predicted that the U-bend waveguide with a 3 mum bend radius can be realized with the propagation loss of < 0.5 dB. PMID- 19547174 TI - Visible three-dimensional metallic photonic crystal with non-localized propagating modes beyond waveguide cutoff. AB - We report experimental realization of a 5-layer three-dimensional (3D) metallic photonic crystal structure that exhibits characteristics of a 3D complete bandgap extending from near-infrared down to visible wavelength at around 650 nm. The structure also exhibits a new kind of non-localized passband mode in the infrared far beyond its metallic waveguide cutoff. This new passband mode is drastically different from the well-known defect mode due to point or line defects. Three dimensional finite-difference-time-domain simulations were carried out and the results suggest that the passband modes are due to intra-structure resonances. PMID- 19547175 TI - All-fiber widely tunable Raman fiber laser with controlled output spectrum. AB - A simple all-fiber widely tunable phosphosilicate Raman fiber laser (RFL) of high efficiency has been developed. The laser has more than 50 nm tuning range, and generates up to 3.2 W of output power with 72% maximum slope efficiency. The output power is almost constant in the range 1258-1303 nm. The width and the spectral power density of the RFL output spectrum can be controlled by the detuning of its cavity fiber Bragg gratings (FBGs) thus being optimized for efficient frequency doubling. PMID- 19547176 TI - All-optical modulation format conversion from on-off-keying to multiple-level phase-shift-keying based on nonlinearity in optical fiber. AB - We propose an all-optical modulation format conversion scheme from non-return-to zero on-off-keying (NRZ-OOK) to return-to-zero (RZ) multiple-level phase-shift keying (PSK) based on nonlinearity in optical fiber. The proposed conversion scheme is numerically investigated and experimentally demonstrated. We experimentally demonstrate error-free operation of NRZ-OOK/RZ- binary PSK conversion at 10.7 Gb/s. The operation of the NRZ-OOK/RZ-quadrature PSK conversion is shown by eye opening after balanced receiving at a symbol rate of 10.7 Gsymbol/s. In addition, we demonstrate the feasibility of the modulation format conversion from NRZ-OOK to RZ-8-levels PSK by numerical simulation. PMID- 19547177 TI - Fabrication and properties of metalo-dielectric photonic crystal structures for infrared spectral region. AB - We report structural and optical properties of three-dimensional periodic metallic woodpile structures obtained by direct laser writing in dielectric photoresist SU-8 and subsequent electroless coating by a thin Ni film. Signatures of photonic stop gaps were observed in optical reflection spectra of the structures at infrared wavelengths. This study demonstrates that the combination of DLW and chemical infiltration of metals is attractive as a simple and cost efficient method for the fabrication of metalo-dielectric photonic crystals. PMID- 19547178 TI - Practical quantum key distribution over 60 hours at an optical fiber distance of 20km using weak and vacuum decoy pulses for enhanced security. AB - Experimental one-way decoy pulse quantum key distribution running continuously for 60 hours is demonstrated over a fiber distance of 20km. We employ a decoy protocol which involves one weak decoy pulse and a vacuum pulse. The obtained secret key rate is on average over 10kbps. This is the highest rate reported using this decoy protocol over this fiber distance and duration. PMID- 19547179 TI - InP-based compact photonic crystal directional coupler with large operation range. AB - We present the design, fabrication and measurement of photonic crystal directional couplers in the InP/InGaAsP/InP material system. A comprehensive analysis of the dependence of the coupling length and usable wavelength range on the diameter of the holes next to the waveguides is given. The possibility to trade-off coupling length against usable wavelength range is shown. Designs with coupling lengths as low as 52 lattice constants and with an operation range covering 16% of the bandgap width are fabricated and measured. Good agreement between optimized and measured devices is achieved. PMID- 19547180 TI - The far-field modified uncorrelated single-scattering approximation in light scattering by a small volume element. AB - Consider light scattering by a small volume element filled with randomly positioned particles, the far-field modified uncorrelated single-scattering approximation (MUSSA) leads to the incoherent summation of the phase matrices of particles in the volume. The validity of the MUSSA is revisited in this paper to include the variation of the particles' positions. Analytical results show that the MUSSA does not require the distance between any pair of particles in the volume to be larger than what is required in the single-scattering approximation (SSA). Instead, it requires the dimension of the volume to be large compared to the incident wavelength. The new results also make the requirements of MUSSA easier to be met. We also analyze energy conservation for the MUSSA. PMID- 19547181 TI - Group delay dispersion measurement of Yb:Gd(2)SiO(5), Yb:GdYSiO(5) and Yb:LuYSiO(5)crystal with white-light interferometry. AB - We report the measured group delay dispersion (GDD) of new crystals Yb:Gd(2)SiO,(5)(Yb:GSO), Yb:GdYSiO(5) (Yb:GYSO) and Yb:LuYSiO(5) (Yb:LYSO) over wavelengths from 1000nm to 1200nm, with a white-light interferometer. Those GDD data should be useful for the dispersion compensation for femtosecond pulse generation in the lasers with these new crystals as the gain media. PMID- 19547182 TI - Control of directional evanescent coupling in fs laser written waveguides: Erratum. AB - An erratum is presented to correct typing mistakes in our paper [Opt. Express 15, 1579 (2007)]. PMID- 19547183 TI - Laser-induced (endo)vascular photothermal effects studied by combined brightfield and fluorescence microscopy in hamster dorsal skin fold venules. AB - The putative features of the (endo)vascular photothermal response, characterized by laser-induced thermal denaturation of blood and vessel wall constituents, have been elucidated individually, but not simultaneously in dynamic, isolated in vivo systems. A hamster dorsal skin fold model in combination with brightfield/fluorescence intravital microscopy was used to examine the effect of laser pulse duration and blood flow velocity on the size of the thermal coagulum, its attachment behavior, and laser-mediated vasomotion. The size of the coagulum and the extent of vasoconstriction and latent vasodilation were proportional to the laser pulse duration, but pulse duration had no effect on coagulum attachment/dislodgement. Blood flow velocity exhibited no significant effect on the studied parameters. The (endo)vascular photothermal response is governed predominantly by laser energy deposition and to a marginal extent by blood flow velocity. PMID- 19547184 TI - Asynchronous Digital Optical Regenerator for 4 x 40 Gbit/s WDM to 160 Gbit/s OTDM Conversion. AB - We propose and numerically analyse an asynchronous digital optical regenerator using a single-EAM loop and a novel neighbor-combine approach. It effectively re synchronizes input signals with arbitrary phases to the local clock, and regenerates signals with high amplitude fluctuation and polarization mode dispersion. We demonstrate the application of this regenerator for 4x40 Gbit/s WDM to 160 Gbit/s OTDM conversion. PMID- 19547185 TI - Optical channel waveguides by proton and carbon implantation in Nd:YAG crystals. AB - In this work the formation of optical channel waveguides in Nd:YAG crystals by either proton or carbon implantation is reported. The channel waveguides were obtained by a single implantation process through an electroformed mask of nickel cobalt alloy. Experimental measurements of the optical properties of these waveguides are presented. PMID- 19547186 TI - Femtosecond laser nanoaxotomy properties and their effect on axonal recovery in C. elegans. AB - We present a study characterizing the properties of femtosecond laser nanosurgery applied to individual axons in live Caenorhabditis elegans (C. elegans) using nano-Joule laser pulses at 1 kHz repetition rate. Emphasis is placed on the characterization of the damage threshold, the extent of damage, and the statistical rates of axonal recovery as a function of laser parameters. The ablation threshold decreases with increasing number of pulses applied during nanoaxotomy. This dependency suggests the existence of an incubation effect. In terms of extent of damage, the energy per pulse is found to be a more critical parameter than the number of pulses. Axonal recovery improves when surgery is performed using a large number of low energy pulses. PMID- 19547187 TI - Topology measurements of metal nanoparticles with 1 nm accuracy by Confocal Interference Scattering Microscopy. AB - We present a novel scattering microscopy method to detect the orientation of individual silver nanorods and to measure their relative distances. Using confocal microscopy in combination with either the fundamental or higher order laser modes, scattering images of silver nanorods were recorded. The distance between two individual nanorods was measured with an accuracy in the order of 1 nm.We detected the orientation of isolated silver nanorods with a precision of 0.5 degree that corresponds to a rotational arch of about 1 nm. The results demonstrate the potential of the technique for the visualization of non-bleaching labels in biosciences. PMID- 19547188 TI - Modified polymethylmethacrylate as a base for thermostable optical recording media. AB - A possibility to improve the thermal properties of holographic gratings in a photosensitive system based on polymethylmethacrylate (PMMA) and to enhance simultaneously the adhesion of the photopolymer to soda-lime glass is demonstrated. The modified PMMA was prepared by radical copolymerisation of methylmethacrylate (MMA) with acrylic acid (AA). Polymer films deposited from samples of the copolymer of MMA with AA containing 9,10-phenanthrenequinone additives were used as a photosensitive material for the recording of holographic gratings. It is possible to generate gratings that are thermally stable up to 200 masculineC using this modified PMMA. Dynamic thermogravimetry, differential thermal analysis and thermal mechanic analyses were used to determine the dependence of the thermal stability of the modified PMMA on the composition and the structure of its macromolecules. PMID- 19547189 TI - Analytical model for quantitative prediction of material contrasts in scattering type near-field optical microscopy. AB - Nanometer-scale mapping of complex optical constants by scattering-type near field microscopy has been suffering from quantitative discrepancies between the theory and experiments. To resolve this problem, a novel analytical model is presented here. The comparison with experimental data demonstrates that the model quantitatively reproduces approach curves on a Au surface and yields an unprecedented agreement with amplitude and phase spectra recorded on a phonon polariton resonant SiC sample. The simple closed-form solution derived here should enable the determination of the local complex dielectric function on an unknown sample, thereby identifying its nanoscale chemical composition, crystal structure and conductivity. PMID- 19547190 TI - Large-area, three-state, binary amplitude and binary phase vertical-cavity multiple quantum well electroabsorption modulator. AB - We present the design and characterization of a large optical modulator array based on GaAs multiple quantum wells for amplitude and phase modulation. The device shows two high-reflectance states with a phase difference close to 180 degrees for use as a binary phase modulator. It also shows a third, low reflectance state for use as an amplitude modulator. It is segmented into 64 pixels in a single row, giving an active area of 2mm x 5mm. We discuss the device performance as a ternary binary amplitude and binary phase modulator, including contrast ratio and uniformity, and show that a voltage swing of only 5V is needed to drive it. PMID- 19547191 TI - Analytical tools for customized design of monofocal intraocular lenses. AB - We propose a complete methodology to develop custom monofocal Intraocular Lens (IOL) designs and evaluate their performance on-axis based on an analytical formulation. The analytical formulation was based on Gaussian and primary aberration theory applied to custom (individual biometric data) and realistic (multilayer cornea and thick IOL) pseudoaphakic eye models. Gradient-based optimization algorithms were performed to search for optimal designs. Using two parameters, the best design was obtained by directly minimizing the wavefront variance. We showed, in a case example, that custom designs achieved better final performance than generic IOL designs. Tolerances analysis allowed an evaluation of the implications of the manufacturing errors of the different parameters. PMID- 19547193 TI - Multi-millijoule, diode-pumped, cryogenically-cooled Yb:KY(WO(4))(2) chirped pulse regenerative amplifier. AB - A diode-pumped, cryogenically-cooled Yb:KYW regenerative amplifier utilizing chirped-pulse amplification and regenerative pulse shaping has been developed. An amplified pulse with an energy of 5.5 mJ and a broad bandwidth of 3.4 nm is achieved. PMID- 19547192 TI - Oil droplets as light absorbents in seawater. AB - This paper presents spectra of light absorption coefficient of oil-in-water emulsion, derived using the Mie theory. In order to achieve that concentration of oil, degree of oil dispersion in seawater as well as spectra of light absorption coefficient and refractive index of chosen oils must be known. A significant role of the size distribution of oil droplets has been revealed: light absorption coefficient of emulsion increases with the rate of dispersion. PMID- 19547194 TI - Luminescence enhancement by Au nanoparticles in Er(3+)-doped germano-silicate optical fiber. AB - We report on the fabrication of the Au nanoparticles/Er(3+) codoped germano silicate fibers by modified chemical vapor deposition and solution doping processes. Absorption and luminescence characteristics of the Er(3+)-doped germano-silicate fibers incorporated with Au nanoparticles in the core of the fibers were investigated. The Au nanoparticles were found to be effective absorbents for hydroxyl groups to enhance the luminescence of Er(3+) ions upon pumping with the 980nm laser diode. PMID- 19547195 TI - Optical film to enhance cosmetic appearance and brightness in liquid crystal displays. AB - This paper presents an optical film that enhances cosmetic appearance as well as brightness in a liquid crystal display (LCD) through microprisms that have a variable pitch. The optical film utilizes Fourier transformation to optimize the arrangement of microprisms for improving the cosmetic appearance in the display. The optical film has an improved light-collimating feature that redirects light more effectively, resulting in higher brightness. This paper shows details of the design procedure, but more importantly, presents optical measurement results of an actual optical film prototype to confirm the performance improvement. PMID- 19547196 TI - Optical ferris wheel for ultracold atoms. AB - We propose a versatile optical ring lattice suitable for trapping cold and quantum degenerate atomic samples. We demonstrate the realisation of intensity patterns from pairs of Laguerre-Gauss (exp(i??) modes with different ? indices. These patterns can be rotated by introducing a frequency shift between the modes. We can generate bright ring lattices for trapping atoms in red-detuned light, and dark ring lattices suitable for trapping atoms with minimal heating in the optical vortices of blue-detuned light. The lattice sites can be joined to form a uniform ring trap, making it ideal for studying persistent currents and the Mott insulator transition in a ring geometry. PMID- 19547197 TI - Multiple wavelength resonant grating filters at oblique incidence with broad angular acceptance. AB - Multilayer, multimode waveguides are utilized in resonant grating filters having a broadened angular acceptance bandwidth for multiple wavelengths at a single oblique angle of incidence. It is shown that the waveguide grating structure should support a few leaky modes in order to support a multiwavelength resonant filter at oblique incidence with broadened angle acceptance at each wavelength. PMID- 19547198 TI - Optics of metal nanoparticle aggregates with light induced motion. AB - Light-induced forces between metal nanoparticles change the geometry of the aggregates and affect their optical properties. Light absorption, scattering and scattering of a probe beam are numerically studied with Newton's equations and the coupled dipole equations for penta-particle aggregates. The relative changes in optical responses are large compared with the linear, low-intensity limit and relatively fast with nanosecond characteristic times. Time and intensity dependencies are shown to be sensitive to the initial potential of the aggregation forces. PMID- 19547199 TI - Fabrication of high-resolution periodical structure on polymer waveguides using a replication process. AB - This paper describes a procedure to replicate a polymeric wavelength filter. In this work, the grating structure on a polymer is fabricated first using holographic interferometry and micro-molding processes. The polymeric wavelength filters are produced by a two-step molding process where the master mold is first formed on a negative tone photoresist and subsequently transferred to a PDMS mold; following this step, the PDMS silicon rubber mold was used as a stamp to transfer the pattern of the polymeric wavelength filters onto a UV cure epoxy. Initial results show good pattern transfer in physical shape. At the Bragg wavelength, a transmission dip of -15.5 dB relative to the -3dB background insertion loss and a 3-dB-transmission bandwidth of ?6nm were obtained from the device. PMID- 19547200 TI - Simultaneous monitoring technique for ASE and MPI noises in distributed Raman Amplified Systems. AB - We develop a new technique for simultaneously monitoring the amplified spontaneous emission (ASE) and multi-path interference (MPI) noises in distributed Raman amplified (DRA) systems. This technique utilizes the facts that the degree-of polarization (DOP) of the MPI noise is 1/9, while the ASE noise is unpolarized. The results show that the proposed technique can accurately monitor both of these noises regardless of the bit rates, modulation formats, and optical signal-to-noise ratio (OSNR) levels of the signals. PMID- 19547201 TI - A coherent framework for fingerprint analysis: are fingerprints Holograms? AB - We propose a coherent mathematical model for human fingerprint images. Fingerprint structure is represented simply as a hologram - namely a phase modulated fringe pattern. The holographic form unifies analysis, classification, matching, compression, and synthesis of fingerprints in a self-consistent formalism. Hologram phase is at the heart of the method; a phase that uniquely decomposes into two parts via the Helmholtz decomposition theorem. Phase also circumvents the infinite frequency singularities that always occur at minutiae. Reliable analysis is possible using a recently discovered two-dimensional demodulator. The parsimony of this model is demonstrated by the reconstruction of a fingerprint image with an extreme compression factor of 239. PMID- 19547202 TI - Editorial: 10th anniversary. AB - July 7 is the tenth anniversary of Optics Express. It started in 1997 under the Editorship, and general championship, of J. H. Eberly and 9 Associate Editors, 8 of whom are from the USA. For comparison: at present there are 41 Associate Editors, 20 of whom are from the USA (more about them later). Around the time that Michael Duncan took over as Editor in 2002, submissions really took off, quadrupling between 2002 and 2004. This was in part because of the large number of papers in Microstructured Optical Fibers, for which, thanks to its rapid time to publication, Optics Express became the journal of choice. Optics Express has now evolved to be the journal of choice of the wider optics community: witness the recently released 2006 impact factor of 4.009, the highest of all optics journals. PMID- 19547203 TI - Development of fluorescent materials for Diffuse Fluorescence Tomography standards and phantoms. AB - The availability of fluorescence standards is necessary in the development of systems and methods for fluorescence imaging. In this study, two approaches for developing diffuse fluorescence materials to be used as standards or phantoms in diffuse fluorescent tomography applications were investigated. Specifically, silicone rubber and polyester casting resin were used as base materials, and silicone pigments or TiO(2) / India Ink were added respectively to vary the optical properties. Characterization of the optical properties achieved was performed using time-resolved methods. Subsequently, different near-infrared fluorochromes were examined for imparting controlled and stable fluorescence properties. It was determined that hydrophobic fluorophores (IR 676 and IR 780 Iodide) suspended in dichloromethane and hydrophilic fluorophores (Cy5.5 and AF 750) suspended in methanol produced diffusive silicone and resin fluorescent materials, respectively. However only the hydrophobic fluorophores embedded within silicone resulted in the construction of a material with the characteristics of a standard, i.e. stability of fluorescence intensity with time and a linear dependence of normalized fluorescence intensity to fluorophore concentration. PMID- 19547204 TI - Multipolar tensor analysis of second-order nonlinear optical response of surface and bulk of glass. AB - We use two-beam second-harmonic generation to perform a quantitative tensor analysis of the effective dipolar surface nonlinearity and the separable multipolar bulk nonlinearity for BK7 glass. The most straightforward, self consistent interpretation of the results is obtained when the effective surface response is assumed to have approximate Kleinman symmetry and the bulk contribution is dominated by magnetic, rather than quadrupole, effects. PMID- 19547205 TI - Target-locking acquisition with real-time confocal (TARC) microscopy. AB - We present a real-time target-locking confocal microscope that follows an object moving along an arbitrary path, even as it simultaneously changes its shape, size and orientation. This Target-locking Acquisition with Realtime Confocal (TARC) microscopy system integrates fast image processing and rapid image acquisition using a Nipkow spinning-disk confocal microscope. The system acquires a 3D stack of images, performs a full structural analysis to locate a feature of interest, moves the sample in response, and then collects the next 3D image stack. In this way, data collection is dynamically adjusted to keep a moving object centered in the field of view. We demonstrate the system's capabilities by target-locking freely-diffusing clusters of attractive colloidal particles, and activelytransported quantum dots (QDs) endocytosed into live cells free to move in three dimensions, for several hours. During this time, both the colloidal clusters and live cells move distances several times the length of the imaging volume. PMID- 19547206 TI - Analysis of linearity of highly saturated electroabsorption modulator link due to photocurrent feedback effect. AB - We have analyzed the linearity performance of analog fiber-optic links based on electroabsorption modulators (EAM) operating at high optical power. The negative feedback caused by photocurrent generation improves the modulator linearity in the gain saturation regime. In the absence of laser relative intensity noise (RIN), the link spur-free dynamic range (SFDR) increases with the power of four thirds of the input optical power after gain saturation occurs. A multi-octave SFDR of more than 135 dB/Hz2/3 has been found to be achievable with sufficiently high power. PMID- 19547207 TI - In-line short cavity Fabry-Perot strain sensor for quasi distributed measurement utilizing standard OTDR. AB - This paper presents an in-line, short cavity Fabry-Perot fiber optic strain sensor. A short air cavity inside a single-mode fiber is created by the fusion splicing of appropriately micro machined fiber tips. A precise tuning of the cavity length is introduced and used for the setting of the sensor static characteristics within the quasi-linear range around a quadrature point, which significantly simplifies signal processing. Sensor insertion losses achieved by short cavity design and optimized fusion splicing proved to be below 1 dB. Low insertion loss allows for effective cascading of the proposed strain sensors into a quasi-distributed sensor array. A practical 10-point quasi-distributed strain sensor array was demonstrated in practice, where each in-line sensor was tuned to the same operating point in the static characteristics, thus allowing for simple interrogation of the sensor array by using standard telecommunication OTDR. In addition, precise tuning of the short cavity Fabry Perot sensor was applied for an effective compensation of temperature-induced strain errors and for an increase in the unambiguous measuring range, while improving the overall linearity of the sensor system. PMID- 19547208 TI - Micro-channels machined in microstructured optical fibers by femtosecond laser. AB - Micro-channels were fabricated in hollow-core photonic bandgap fiber (HC-PBGF) and suspended-core holey fiber (SC-HF) by femtosecond Ti:sapphire laser irradiation. Gaseous access was demonstrated via these engineered ports to the core of HC-PBGF and the hollow cladding of SC-HF. Femtosecond laser micro machining caused no additional transmission loss in HC-PBGFs. This allowed a novel gas cell to be produced, in which gaseous access was provided solely through two micro-channels. Acetylene diffusion was also confirmed through a micro-channel leading to a single cladding airhole in SC-HF. This further highlighted the fabrication technique's precision, selectivity, and potential for developing fiber-based micro-fluidic devices. PMID- 19547209 TI - Infrared transmission resonances in double-layered, complementary-structure metallic gratings. AB - A double-layered metallic grating (metal-dielectric-metal) with a complementary capacitive (isolated discs) / inductive (connected film with apertures) structure exhibits multiple infrared transmission resonances peaks with up to 70% at wavelength ranges corresponding to local modes for geometric dimensions less than a wavelength. The period, dielectric thickness, refractive index and unit cell size of the periodic structure modulate the local mode positions and amplitudes. The electromagnetic field distribution and energy flow in the structure explain the relation of transmission resonance, local modes, and distributed surface plasma wave modes. PMID- 19547210 TI - Noise in cavity ring-down spectroscopy caused by transverse mode coupling. AB - In our continuous wave cavity ring-down spectroscopy (CW-CRDS) experiments, we have often observed that the decay time constant drops to a lower value at some cavity lengths or some intercavity pressures. The resulting instabilities lead to a reduction in the sensitivity of our CRDS system. We have deduced that the cause of this noise is the coupling between the TEM(00) mode that the laser excites, and the higher order transverse modes of the cavity. The coupling will cause anti crossings as the modes tune with cavity length. A consequence is that the decay of light intensity leaving the cavity is no longer a single exponential decay, but the signal can be quantitatively fit to a two-mode beating model. With a 4mm diameter intra-cavity aperture, the higher order modes are suppressed and the stability of the system improved greatly. One coupling mechanism is scattering from the mirror surfaces. This can explain some features of our data including the strength of this coupling and the relative tuning rate of the coupled modes. Remarkably, a scattering intensity between modes of ~ 10(-12) can produce observable changes in the cavity decay rate. However, the tuning rate between the TEM(00) mode and the higher order modes in a cavity pressure scan is larger than predicted and is still not explained. Images of higher order transverse modes excited at certain cavity conditions were recorded by an Indium Gallium Arsenide (InGaAs) area camera. PMID- 19547211 TI - Broadband sum-frequency generation as an efficient two-photon detector for optical tomography. AB - We describe a novel non-linear detection method for optical tomography that does not rely on detection of interference fringes and is free of optical background. The method exploits temporally coherent broadband illumination such as ultrashort pulses, and a non-linear two-photon detection process such as sum-frequency generation (SFG). At the detection stage, the reference beam and the sample beam are mixed in a thick non-linear crystal, and only the mixing term, which is free of optical background, is detected. Consequently, the noise limitations posed by the background in standard OCT (excess and shot noise), do not exist here. Due to the non-linearity, the signal to noise ratio scales more favorably with the optical power compared to standard OCT, yielding an inherent improvement for high speed tomographic scans. Careful design of phase matching in the crystal enables non-linear mixing which is both highly efficient and broadband, yielding both high sensitivity and high depth resolution. PMID- 19547212 TI - Spontaneous parametric down conversion in a nanophotonic waveguide. AB - Recently, we verified that spontaneous parametric down conversion (SPDC) is enhanced in a waveguide, in agreement with theory showing an inverse dependence on mode confinement [1]. Here we investigate highly-confined nanophotonic waveguides designed to maximize the SPDC rate. A theory modified to include highly-confined waveguides is used to calculate the spectral width and pair generation rates in a sample system. Pair generation rates exceeding 10(9)/sec/nm/mW are predicted for periodically-poled KTP (PPKTP) nanophotonic waveguides. This results in an enhancement of the downconverted signal power greater than 45x that of low-index-contrast PPKTP waveguides and greater than 6500x that of bulk PPKTP crystals. PMID- 19547213 TI - Dark-bright soliton pairs in nonlocal nonlinear media. AB - We study the formation of dark-bright vector soliton pairs in nonlocal Kerr-type nonlinear medium. We show, by analytical analysis and direct numerical calculation, that in addition to stabilize of vector soliton pairs nonlocal nonlinearity also helps to reduce the threshold power for forming a guided bright soliton. With help of the nonlocality, it is expected that the observation of dark-bright vector soliton pairs in experiments becomes more workable. PMID- 19547214 TI - A novel operating principle in speckle interferometry: the double-focusing. AB - The present paper describes in details the operating principle of a completely new family of speckle interferometers: the double-focusing. This type of interferometer is sensitive to the same components of displacement given by holographic interferometry, i.e. the component along the bisector of the angle identified by the illumination and the observation directions. In addition, no external reference beam is necessary, with a consequent reduction of the complexity of the experimental setup. The only requirement for the correct functioning of this family of interferometers is that only a portion of the illuminated area undergoes a sensible deformation. The implementation can be indifferently carried out by adopting the classical Michelson or Mach-Zender configurations, but also a particularly compact in-line implementation can be realized. PMID- 19547215 TI - Classical dispersion-cancellation interferometry. AB - Even-order dispersion cancellation, an effect previously identified with frequency-entangled photons, is demonstrated experimentally for the first time with a linear, classical interferometer. A combination of a broad bandwidth laser and a high resolution spectrometer was used to measure the intensity correlations between anti-correlated optical frequencies. Only 14% broadening of the correlation signal is observed when significant material dispersion, enough to broaden the regular interferogram by 4250%, is introduced into one arm of the interferometer. PMID- 19547216 TI - Time resolved confocal luminescence investigations on Reverse Proton Exchange Nd:LiNbO(3) channel waveguides. AB - In this work we report on the time and spatial resolved fluorescence of Neodymium ions in LiNbO(3) channel waveguides fabricated by Reverse Proton Exchange. The analysis of the fluorescence decay curves obtained with a sub-micrometric resolution has evidenced the presence of a relevant fluorescence quenching inside the channel waveguide. From the comparison between diffusion simulations and the spatial dependence of the (4)F(3/2) fluorescence decay rate we have concluded that the observed fluorescence quenching can be unequivocally related to the presence of H+ ions in the LiNbO(3) lattice. Nevertheless, it turns out that Reverse Proton Exchange guarantees a fluorescence quenching level significantly lower than in similar configurations based on Proton Exchange waveguides. This fluorescence quenching has been found to be accompanied by a relevant red-shift of the (4)F(3/2)?(4)I(9/2) fluorescence band. PMID- 19547217 TI - Spectral Talbot effect in sampled fiber Bragg gratings with super-periodic structures. AB - Due to the equivalence between an integral multiple of 2pi?and zero in the phase space, a general configuration of sampled fiber Bragg gratings (SFBGs) with super periodic structures has been introduced and investigated. These super-periodic structures can be used to implement spectral Talbot effect with a large degree of freedom, as long as any one of the three parameters involved in quadratic phase profile is an even number. Then the phase profiles of such SFBGs are analyzed in detail. Although their phase increments are constant or non-constant periodic functions in different cases, theoretical analysis and simulations show that the obtained filtering characteristics are the same. In contrast to uniform SFBGs with identical sampling period, multiplied filtering channels and similar groupdelay characteristic are achieved for these SFBGs with super-periodic structures. PMID- 19547219 TI - Effective recording of dynamic phase gratings in Yb-doped fibers with saturable absorption at 1064nm. AB - Results of investigation of transient two-wave mixing via dynamic population gratings in Yb-doped fibers with saturable absorption are reported. The recorded gratings are characterized by submillisecond formation times and at wavelength lambda = 1064 nm need cw recording light power of 1 - 10 mW scale. What is important for different applications, the dynamic gratings were found to be predominantly of a phase type with an admixture of a significantly weaker amplitude component only. As in Er-doped fibers, the amplitude grating component proved to be essentially inferior to the theoretical estimation based on the fiber optical density and saturation power. PMID- 19547218 TI - Influence of shot noise on phase measurement accuracy in digital holographic microscopy. AB - Digital Holographic Microscopy (DHM) is a single shot interferometric technique, which provides quantitative phase images with subwavelength axial accuracy. A short hologram acquisition time (down to microseconds), allows DHM to offer a reduced sensitivity to vibrations, and real time observation is achievable thanks to present performances of personal computers and charge coupled devices (CCDs). Fast dynamic imaging at low-light level involves few photons, requiring proper camera settings (integration time and gain of the CCD; power of the light source) to minimize the influence of shot noise on the hologram when the highest phase accuracy is aimed. With simulated and experimental data, a systematic analysis of the fundamental shot noise influence on phase accuracy in DHM is presented. PMID- 19547220 TI - Millimeter wave probing of the acoustic phase for concealed object detection. AB - We developed a far infrared technique for detecting objects that are concealed behind a barrier, which is opaque for visible light. The technique makes use of the fact that mechanically coupled objects can be distinguished by their acoustic phase during a driven vibration. The pattern of the acoustic phase is imaged using 70 GHz radiation. The experimental data demonstrate that virtually all objects can be sensed, which includes metals as well as dielectric materials. PMID- 19547221 TI - Thermal response of Bragg gratings in PMMA microstructured optical fibers. AB - We report on the thermal characteristics of Bragg gratings fabricated in polymer optical fibers. We have observed a permanent shift in the grating wavelength at room temperature which occurs when the grating has been heated above a threshold temperature. This threshold temperature is dependent on the thermal history of the grating, and we attribute the effect to a shrinking of the fiber. This effect can be avoided by annealing the fiber before grating inscription, resulting in a linear response with temperature and an increased linear operating temperature range of the grating. PMID- 19547222 TI - Numerical suppression of zero-order image in digital holography. AB - This work describes a novel approach that adopts numerical operation to suppress the zero-order images of reconstruction in digital holography. The entire process needs only one digital hologram and keeps under control the intensity ratio of the object wave to reference wave in recording procedure. Also the performance of numerical suppression is simple and effective by subtracting the numerical generated intensity of the object and reference waves from the digital hologram. The experimental results demonstrate that the zero-order images of reconstruction can be suppressed completely and represents the satisfactory reconstructed image even if the distribution of the object wave is not uniform. Therefore this approach can simplify the procedure of phase-shifting digital holographic-based scheme involving multiple exposures. Moreover, the investigation of performance using the novel suppression approach is presented for proving the practical feasibility. PMID- 19547223 TI - Thermal sensitivity of tellurite and germanate optical fibers. AB - The temperature coefficients of optical phase have been measured at 1536 nm wavelength for short fiber Fabry-Perot cavities of tellurite and germanate glass fibers spliced to silica fiber. The results are consistent with the thermal expansion and thermo-optic coefficients of the bulk glasses. PMID- 19547224 TI - Femtosecond Ti:sapphire cryogenic amplifier with high gain and MHz repetition rate. AB - We demonstrate high gain amplification of 160-femtosecond pulses in a compact double-pass cryogenic Ti:sapphire amplifier. The setup involves a negative GVD mirrors recompression stage, and operates with a repetition rate between 0.2 and 4 MHz with a continuous pump laser. Amplification factors as high as 17 and 320 nJ Fourier-limited pulses are obtained at a 800 kHz repetition rate. PMID- 19547225 TI - The origin of magnetic polarizability in metamaterials at optical frequencies - an electrodynamic approach. AB - We explain the origin of the electric and particular the magnetic polarizabiltiy of metamaterials employing a fully electromagnetic plasmonic picture. As example we study an U-shaped split-ring resonator based metamaterial at optical frequencies. The relevance of the split-ring resonator orientation relative to the illuminating field for obtaining a strong magnetic response is outlined. We reveal higher-order magnetic resonances and explain their origin on the basis of higher-order plasmonic eigenmodes caused by an appropriate current flow in the split-ring resonator. Finally, the conditions required for obtaining a negative index at optical frequencies in a metamaterial consisting of split-ring resonators and wires are investigated. PMID- 19547226 TI - Narrow-bandwidth picosecond pulses by spectral compression of femtosecond pulses in second-order nonlinear crystals. AB - We introduce a simple approach for the efficient generation of tunable narrow bandwidth picosecond pulses synchronized to broadband femtosecond ones. Second harmonic generation in the presence of large group velocity mismatch between the interacting pulses transfers a large fraction of the energy of a broadband fundamental frequency pulse into a narrowband second harmonic one. Using a periodically poled stoichiometric lithium tantalate crystal coupled to an infrared optical parametric amplifier, we generated 200-nJ pulses with spectral width lower than 8.5 cm(-1) and tunability from 720 to 890 nm. Energy scaling and extension of the tuning range are straightforward. PMID- 19547227 TI - Highly birefringent elliptical-hole rectangular-lattice photonic crystal fibers with modified air holes near the core. AB - In this work, modal birefringence and polarization-dependent leakage loss for the fundamental mode in elliptical-hole rectangular-lattice photonic crystal fibers (EH-RL-PCFs) with modified air holes near the core are investigated by using a full-vector, finite-difference method in the frequency domain. Numerical results show that by introducing modified air holes, a heightened modal birefringence of the order of 10(-2) has been obtained within the wavelengths ranging from 1.0 to 1.6 mum in the proposed PCFs, with a lattice length of 2 mum and with a large lattice length of 3 mum at1.55 mum. PMID- 19547228 TI - Fast one-dimensional photonic crystal modulators for the terahertz range. AB - Optically controlled one-dimensional photonic crystal structures for the THz range are studied both theoretically and experimentally. A GaAs:Cr layer constitutes a defect in the photonic crystals studied; its photoexcitation by 800 nm optical femtosecond pulses leads to the modulation of the THz beam. Since the THz field can be localized in the photoexcited layer of the photonic crystal, the interaction between photocarriers and THz light is strengthened and yields an appreciable modulation of the THz output beam even for low optical pump fluences. Optimum resonant structures are found, constructed and experimentally studied. The dynamical response of these elements is shown to be controlled by the lifetime of THz photons in the resonator and by the free carrier lifetime. The time response of the structures studied is shorter than 330 ps. PMID- 19547229 TI - Signal epidetection in third-harmonic generation microscopy of turbid media. AB - Third-harmonic generation (THG) imaging of thick samples or large organisms requires TH light to be epicollected through the focusing objective. In this study we first estimate the amount of backward-to-forward TH radiation created by an isolated object as a function of size and spatial frequencies in the object. Theory and model experiments indicate that no significant signal can be epidetected from a (biological) dielectric structure embedded in a transparent medium. In contrast, backward emission is observed from metal nanoparticles where THG is partly a surface effect. We then address the case of an object embedded in a turbid medium. Experiments and Monte Carlo simulations show that epidetection is possible when the absorption mean free path of harmonic light in the medium exceeds its reduced scattering length, and that epicollection efficiency critically depends on the microscope field-of-view even at shallow depths, because backscattered light is essentially diffusive. These observations provide guidelines for optimizing epidetection in third-harmonic, second-harmonic, or CARS imaging of thick tissues. PMID- 19547230 TI - Directional couplers operated by resonant coupling in all-solid photonic bandgap fibers. AB - We have proposed directional couplers operated by resonant coupling in all-solid photonic bandgap fibers structure which consist of a cladding with an array of high-index rods in silica background, two cores formed by omitting two rods, and some defect rods introduced by reducing the diameter of the high-index rods between the cores. The resonant effect induced by the avoided crossing between core-guided supermodes and defect-guided modes significant decreases the coupling length. The directional couplers proposed in this paper are almost polarization independent and have potential application in realizing integrating all-fiber communication devices. PMID- 19547231 TI - Pulse source based on directly modulated laser and phase modulator. AB - We propose the simple pulse source, in which the pulses generated by large-signal directly modulated laser diode are phase-modulated, and then compressed into short pulses by an optimized length of DCF. On the one hand, phase modulator is used to enhance the negative chirp of large-signal directly modulated pulses. On the other hand, the largesignal directly modulated pulses are used to suppress the pedestal produced by the phase modulator. Using this technique, highly stable 10-GHz 5.5-ps optical pulses are obtained, which suppress pedestal up to 20dB and have a low timing jitter of 184fs. PMID- 19547232 TI - Color center conversion by femtosecond pulse laser irradiation in LiF:F(2) crystals. AB - We report the observation of intense spontaneous emission of green light from LiF:F(2):F(3) (+) centers in active channel waveguides generated in lithium fluoride crystals by near-infrared femtosecond laser radiation. While irradiating the crystal at room temperature with 405 nm light from a laser diode, yellow and green emission was seen by the naked eye. Stripe waveguides were fabricated by translating the crystal along the irradiated laser pulse, and their guiding properties and fluorescence spectra at 540 nm demonstrated. This single-step process inducing a waveguide structure offers a good prospect for the development of a waveguide laser in bulk LiF crystals. PMID- 19547233 TI - Emission characteristics of ion-irradiated In(0.53)Ga(0.47)As based photoconductive antennas excited at 1.55 microm. AB - We present a detailed study of the effect of the carrier lifetime on the terahertz signal characteristics emitted by Br(+)-irradiated In(0.53)Ga(0.47)As photoconductive antennas excited by 1550 nm wavelength femtosecond optical pulses. The temporal waveforms and the average radiated powers for various carrier lifetimes are experimentally analyzed and compared to predictions of analytical models of charge transport. Improvements in bandwidth and in average power of the emitted terahertz radiation are observed with the decrease of the carrier lifetime on the emitter. The power radiated by ion-irradiated In(0.53)Ga(0.47)As photoconductive antennas excited by 1550 nm wavelength optical pulses is measured to be 0.8 muW. This value is comparable with or greater than that emitted by similar low temperature grown GaAs photoconductive antennas excited by 780 nm wavelength optical pulses. PMID- 19547234 TI - Long-term stable microwave signal extraction from mode-locked lasers. AB - Long-term synchronization between two 10.225 GHz microwave signals at +10 dBm power level, locked to a 44.26 MHz repetition rate passively mode-locked fiber laser, is demonstrated using balanced optical-microwave phase detectors. The out of-loop measurement result shows 12.8 fs relative timing jitter integrated from 10 Hz to 10 MHz. Long-term timing drift measurement shows 48 fs maximum deviation over one hour, mainly limited by drift of the out-of-loop characterization setup itself. To the best of our knowledge, this is the first time to demonstrate long term (>1 hour) 3 mrad-level phase stability of a 10.225 GHz microwave signal extracted from a mode-locked laser. PMID- 19547235 TI - On the rotational stability of nonspherical particles driven by the radiation torque. AB - We calculate the radiation torque exerted by a monochromatic plane wave, either unpolarized or linearly polarized, on aggregates of spheres and investigate the stability of the resulting rotational motion. In fact, neglecting any braking momenta we calculate the component of the electromagnetic torque orthogonal to the principal axis of maximum moment of inertia through the center of mass (transverse torque), as a function of the direction of propagation of the incident field. The aggregates we study are composed of homogeneous spheres, possibly of different materials. The electromagnetic torque is calculated through the transition matrix approach along the lines of the theory reported in our recent paper [F. Borghese, P. Denti, R. Saija and M. A. Iati, Opt. Express 14, 9508 (2006)]. When the transverse component of the electromagnetic torque is small or vanishes the rotational motion driven by the component along the principal axis of inertia may be nearly stable. PMID- 19547236 TI - Prediction of the limit of detection of an optical resonant reflection biosensor. AB - A prediction of the limit of detection of an optical resonant reflection biosensor is presented. An optical resonant reflection biosensor using a guided mode resonance filter is one of the most promising label-free optical immunosensors due to a sharp reflectance peak and a high sensitivity to the changes of optical path length. We have simulated this type of biosensor using rigorous coupled wave theory to calculate the limit of detection of the thickness of the target protein layer. Theoretically, our biosensor has an estimated ability to detect thickness change approximately the size of typical antigen proteins. We have also investigated the effects of the absorption and divergence of the incident light on the detection ability of the biosensor. PMID- 19547237 TI - Ultrafast phase and amplitude pulse shaping with a single, one-dimensional, high resolution phase mask. AB - An ultrafast pulse shaper, capable of both phase and amplitude shaping, is constructed using a single high-resolution liquid crystal phase mask. The shaper is calibrated with an inline spectral interferometry technique. Amplitude shaping is accomplished by writing to the mask a phase grating, whose period is smaller than the spectral focus, diffracting away selected frequencies in a controllable manner. PMID- 19547238 TI - Air-clad fibers: pump absorption assisted by chaotic wave dynamics? AB - Wave chaos is a concept which has already proved its practical usefulness in design of double-clad fibers for cladding-pumped fiber lasers and fiber amplifiers. In general, classically chaotic geometries will favor strong pump absorption and we address the extent of chaotic wave dynamics in typical air-clad geometries. While air-clad structures supporting sup-wavelength convex air-glass interfaces (viewed from the high-index side) will promote chaotic dynamics we find guidance of regular whispering-gallery modes in air-clad structures resembling an overall cylindrical symmetry. Highly symmetric air-clad structures may thus suppress the pump-absorption efficiency eta below the ergodic scaling law etainfinity Ac/Acl, where Ac and Acl are the areas of the rare-earth doped core and the cladding, respectively. PMID- 19547239 TI - Impairments in deeply-saturated optical parametric amplifiers for amplitude- and phase-modulated signals. AB - We measure impairment of on-off-keyed and differential-phase-shift-keyed signals imposed by gain saturation in a fiber parametric amplifier. Phase modulation is observed to be more robust, particularly for deep (15 dB) saturation. PMID- 19547240 TI - 2D optical manipulation and assembly of shape-complementary planar microstructures. AB - Optical trapping and manipulation offer great flexibility as a non-contact microassembly tool. Its application to the assembly of microscale building blocks may open new doors for micromachine technology. In this work, we demonstrate all optical assembly of microscopic puzzle pieces in a fluidic environment using programmable arrays of trapping beams. Identical shape-complimentary pieces are optically fabricated with submicron resolution using two-photon polymerization (2PP) technique. These are efficiently assembled into space-filling tessellations by a multiple-beam optical micromanipulation system. The flexibility of the system allows us to demonstrate both user-interactive and computer-automated modes of serial and parallel assembly of microscale objects with high spatial and angular positioning precision. PMID- 19547241 TI - Variable splitting ratio 2 x 2 MMI couplers using multimode waveguide holograms. AB - Variable power splitting ratio 2x2 MMI couplers using multimode waveguide holograms are analyzed. Theoretical analysis shows that variable splitting ratios can be obtained with surface relief holograms on MMI couplers with fixed dimensions. Devices with paired-imaging lengths are designed on a silicon-on insulator (SOI) platform. Beam propagation simulations are used to verify a matrix theory analysis and to investigate proposed device performance. Fabrication tolerance of the proposed device is also analyzed. PMID- 19547242 TI - Coupling terahertz radiation onto a metal wire using a subwavelength coaxial aperture. AB - We demonstrate a simple technique for coupling freely propagating broadband THz radiation to multi-cycle THz pulses on a cylindrical metal wire. This is accomplished by inserting the tapered end of a cylindrical wire into the center of a subwavelength circular aperture fabricated in a freestanding metal film, forming an effective coaxial waveguide. By doing so, we convert the transmission properties of THz pulses through the aperture from an evanescent mode to a propagating mode. By fabricating concentric annular grooves about the aperture, multicycle THz pulses are coupled to the wire. The individual groove geometry, number of grooves, and groove spacing surrounding the subwavelength aperture on the metal film determine the shape of THz pulses launched on the waveguide. PMID- 19547243 TI - All-optical nano modulator on a silicon chip. AB - We present an all-optical modulator realized on a silicon chip. The proposed modulator has nano scale dimensions and a high extinction ratio. Its operation principle is based on a spatially non-uniform variation of the absorption of a miniaturized, silicon waveguide - based Mach-Zehnder interferometer (MZI). The absorption variation is obtained by illuminating the MZI with visible light. Our modulator may be used as an interfacing link between microelectronic processing circuits and optical information transmission links. We provide details on the fabrication and the experimental characterization of the suggested device. Since the operation principle is not based on a high Finesse resonator, the modulator is less sensitive to wavelength changes and its operation rate is not connected to the time required for the optical response to reach steady state but rather to material related effects. PMID- 19547244 TI - Numerical study of nonlinear interactions in a multimode waveguide. AB - Multimode nonlinear pulse propagation within a Ta(2)O(5) rectangular rib waveguide has been numerically simulated. The study provides information relating to both the localized spectral evolution along the waveguide and the transverse spectral variation across the guide. The results explain measurements from our previous near-field scanning microscopy experiments that were designed to map continuum generation along and across such waveguides, and that deviated significantly from simple theory. The simulations predict an increased nonlinear phase modulation compared to that occurring in nonlinear single-mode waveguides, due to intermodal nonlinear effects such as cross-phase modulation, leading to an enhanced spectral broadening. PMID- 19547245 TI - Numerical study on surface plasmon polariton behaviors in periodic metal dielectric structures using a plane-wave-assisted boundary integral-equation method. AB - A novel hybrid technique based on the boundary integral-equation method is proposed for studying the surface plasmon polariton behaviors in two-dimensional periodic structures. Considering the periodicity property of the problem, we use the plane-wave expansion concept and the periodic boundary condition instead of using the periodic Green's function. The diffraction efficiency can then be readily calculated once the equivalent electric and magnetic currents are solved that avoids invoking the numerical calculation of the radiation integral. The numerical validity is verified with the cases of highly conducting materials and practical metals. Numerical convergence can be easily achieved even in the case of a large incident angle as 80o. Based on the numerical scheme, a metal dielectric wavy structure is designed for enhancing the transmittance of optical signal through the structure. The excitation of the coupled surface plasmon polaritons for the high transmission is demonstrated. PMID- 19547246 TI - Towards a nonequilibrium thermodynamic description of incoherent nonlinear optics. AB -

Focus Serial: Frontiers of Nonlinear Optics

This concise review is aimed at providing an introduction to the kinetic theory of partially coherent optical waves propagating in nonlinear media. The subject of incoherent nonlinear optics received a renewed interest since the first experimental demonstration of incoherent solitons in slowly responding photorefractive crystals. Several theories have been successfully developed to provide a detailed description of the novel dynamical features inherent to partially coherent nonlinear optical waves. However, such theories leave unanswered the following important question: Which is the long term (spatiotemporal) evolution of a partially incoherent optical field propagating in a nonlinear medium? In complete analogy with kinetic gas theory, one may expect that the incoherent field may evolve, owing to nonlinearity, towards a thermodynamic equilibrium state. Weak-turbulence theory is shown to describe the essential properties of this irreversible process of thermal wave relaxation to equilibrium. Precisely, the theory describes an irreversible evolution of the spectrum of the field towards a thermodynamic equilibrium state. The irreversible behavior is expressed through the H-theorem of entropy growth, whose origin is analogous to the celebrated Boltzmann's H theorem of kinetic gas theory. It is shown that thermal wave relaxation to equilibrium may be characterized by the existence of a genuine condensation process, whose thermodynamic properties are analogous to those of Bose-Einstein condensation, despite the fact that the considered optical wave is completely classical. In spite of the formal reversibility of optical wave propagation, the condensation process occurs by means of an irreversible evolution of the field towards a homogeneous plane-wave (condensate) with small-scale fluctuations superimposed (uncondensed particles), which store the information necessary for the reversible propagation. As a remarkable result, an increase of entropy ("disorder") in the optical field requires the generation of a coherent structure (plane-wave). We show that, beyond the standard thermodynamic limit, wave condensation also occurs in two spatial dimensions. The numerical simulations are in quantitative agreement with the kinetic wave theory, without any adjustable parameter. PMID- 19547247 TI - Effects of parallel and orthogonal polarization on nonlinear optical characteristics of a 1550 nm VCSOA. AB - Experimental observation of Optical Bistability (OB) and nonlinear gain is reported in a 1550-nm Vertical Cavity Semiconductor Optical Amplifier (VCSOA) under parallel and orthogonal polarized optical injection into the two orthogonal polarizations of the fundamental mode. Different nonlinear switching mechanisms, including anticlockwise and clockwise nonlinear gain and bistability, have been found when the polarization of the externally injected signal matches that of the injected mode, whilst a linear response is measured when the polarization is orthogonal to that of the mode under injection. This diversity of behavior with input polarization offers promise for the potential use of VCSOAs for all-optical signal processing and all-optical switching/routing applications. PMID- 19547248 TI - Amplitude and phase noise sensitivity of modelocked Ti:sapphire lasers in terms of a complex noise transfer function. AB - The amplitude and envelope phase noise of a modelocked laser are shown to depend directly on the pump laser amplitude stability. We characterize the sensitivity of this process by a noise transfer function which represents the complex amplitude-to-amplitude modulation (AM-AM) and amplitude-to-phase modulation (AM PM) conversion gain of the pump-induced amplitude and phase noise, respectively. We find that a linearized laser model extrapolated from relaxation oscillation theory, combined with a thermal model, adequately describe the principal features of the response from <1 Hz to 10 MHz. PMID- 19547249 TI - Analysis on dynamic characteristics of semiconductor optical amplifiers with certain facet reflection based on detailed wideband model. AB - Dynamic characteristics of semiconductor optical amplifiers (SOAs) with certain facet reflection in different operation conditions are theoretically investigated with a detailed wideband model. Influences of different facets reflectivities are numerically simulated for different lengths of active regions. The results indicate that the gain recovery time can be reduced to 50% of the initial value while the other related characteristics are optimized for appropriate facets reflections. A half reflective semiconductor optical amplifier (HR-SOA) with a cleaved facet on rear facet and an antireflection coating on front facet can speed up the gain recovery with easy realization and low cost. The related characteristics of this structure are evaluated. It's also indicated that the gain recovery has further potential to be reduced as low as twenties picoseconds for a long active region. PMID- 19547250 TI - Non-classical light emission from a single electrically driven quantum dot. AB - Easy to handle light sources with non-classical emission features are strongly demanded in the growing field of quantum communication. We report on single photon emission from an electrically pumped quantum dot with unmatched spectral purity, making spatial or spectral filtering dispensable. PMID- 19547251 TI - Fiber-optic confocal microscope using a MEMS scanner and miniature objective lens. AB - We designed and constructed a single-fiber-optic confocal microscope (SFCM) with a microelectromechanical system (MEMS) scanner and a miniature objective lens. Axial and lateral resolution values for the system were experimentally measured to be 9.55 mum and 0.83 mum respectively, in good agreement with theoretical predictions. Reflectance images were acquired at a rate of 8 frames per second, over a 140 mum x 70 mum field-of-view. In anticipation of future applications in oral cancer detection, we imaged ex vivo and in vivo human oral tissue with the SFCM, demonstrating the ability of the system to resolve cellular detail. PMID- 19547252 TI - MW ps pulse generation at sub-MHz repetition rates from a phase conjugate Nd:YVO(4) bounce amplifier. AB - We demonstrated high-repetition-rate (sub-MHz) MW pulse generation by combining a picosecond phase conjugate laser system based on a diode-side-pumped Nd:YVO(4) bounce amplifier with a pulse selector based on a RbTiOPO(4) electro-optical modulator. Peak output powers in the range of 2.8-6.8 MW at a pulse repetition frequency range of 0.33-1.0 MHz were achieved at an extraction efficiency of 34 35%. PMID- 19547253 TI - Near-field characterization of extraordinary optical transmission in sub wavelength aperture arrays. AB - Extra ordinary transmission through arrays of subwavelength apertures has been investigated using near-field scanning optical microscopy. For such studies arrays were fabricated to give maximum resonance enhancement of light transmission at the wavelength of illumination that was used (532 nm). To define this enhancement a design was employed that allowed in one field of view of a near-field image the investigation of single apertures of dimension that was similar to what was incorporated into the sub-wavelength hole array. Significant asymmetry in the transmission and the propagation of the light along the aperture array was detected. This non-uniformity could be explained by polarization of the incident light, edge effects and the geometry of the array. The results support a hypothesis of both enhanced transmission due to surface plasmons and a non diffracting beaming as a function of distance effect in the propagation of the light from the array. PMID- 19547254 TI - Analysis of biomolecule detection with optofluidic ring resonator sensors. AB - We theoretically and experimentally analyze the biomolecule detection capability of the liquid core optical ring resonator (LCORR) as a label-free bio/chemical sensor. We first establish a simple and general linear relationship between the LCORR's bulk refractive index sensitivity (BRIS) and its response to molecule deposition onto the surface, which enables us to easily characterize the LCORR sensing performance. Then, biosensing experiments are performed with bovine serum albumin (BSA) and LCORRs of various BRISs. The experimental results are in good agreement with the theoretical prediction. Further analysis shows that the LCORR is capable of detecting BSA below 10 pM with sub-picogram/mm2 mass detection limit. PMID- 19547255 TI - Supermode Si/III-V hybrid lasers, optical amplifiers and modulators: A proposal and analysis. AB - We describe a hybrid laser structure which consists of an amplifying III-V waveguide proximity-coupled to a passive Si waveguide. By operating near the synchronism point (where the phase velocities of the individual waveguides are equal), we can cause the optical power to be confined to any of the two waveguides. This is accomplished by control of waveguides' geometry. In the portion of the supermode resonator where amplification takes place, the mode is confined nearly completely to III-V guide thus realizing a near maximal gain. Near the output facet, the mode power is confined to the Si waveguide thus optimizing the output coupling. This is to be contrasted with approaches which depend on evanescent field penetration into the III-V medium to obtain gain. PMID- 19547256 TI - AlF(3) thin films deposited by reactive magnetron sputtering with Al target. AB - Aluminum fluoride thin films have been deposited by magnetron sputtering of an aluminum target with CF(4) , and CF(4) mixed O(2) as the working gas onto a room temperature substrate. The quality of the coated AlF(3) film applied with 25W sputtering power using CF(4) mixed 5% O(2) was better than for films deposited using conventional methods. The extinction coefficient of AlF(3) was smaller than 6.0x10(-4) in the wavelength range of 190nm to 250nm. Single layer antireflection coatings on both sides of a fused silica substrate increased the transmittance from less than 91% for a bare substrate to higher than 96% in the wavelength range between 190nm to 250nm. PMID- 19547257 TI - Quantification of optical Doppler broadening and optical path lengths of multiply scattered light by phase modulated low coherence interferometry. AB - We show experimental validation of a novel technique to measure optical path length distributions and path length resolved Doppler broadening in turbid media for different reduced scattering coefficients and anisotropies. The technique involves a phase modulated low coherence Mach-Zehnder interferometer, with separate fibers for illumination and detection. Water suspensions of Polystyrene microspheres with high scattering and low absorption levels are used as calibrated scattering phantoms. The path length dependent diffusion broadening or Doppler broadening of scattered light is shown to agree with Diffusive Wave Spectroscopy within 5%. The optical path lengths are determined experimentally from the zero order moment of the phase modulation peak around the modulation frequency in the power spectrum and the results are validated with Monte Carlo simulations. PMID- 19547258 TI - A new mechanism for negative refraction and focusing using selective diffraction from surface corrugation. AB - Refraction at a smooth interface is accompanied by momentum transfer normal to the interface. We show that corrugating an initially smooth, totally reflecting, non-metallic interface provides a momentum kick parallel to the surface, which can be used to refract light negatively or positively. This new mechanism of negative refraction is demonstrated by visible light and microwave experiments on grisms (grating-prisms). Single-beam all-angle-negative-refraction is achieved by incorporating a surface grating on a flat multilayered material. This negative refraction mechanism is used to create a new optical device, a grating lens. A planoconcave grating lens is demonstrated to focus plane microwaves to a point image. These results show that customized surface engineering can be used to achieve negative refraction even though the bulk material has positive refractive index. The surface periodicity provides a tunable parameter to control beam propagation leading to novel optical and microwave devices. PMID- 19547259 TI - Optically driven deposition of single-walled carbon-nanotube saturable absorbers on optical fiber end-faces. AB - Optical radiation propagating in a fiber is used to deposit commercially available, single-walled carbon nanotubes on cleaved optical fiber end faces and fiber connectors. Thermophoresis caused by heating due to optical absorption is considered to be a likely candidate responsible for the deposition process. Single-walled carbon nanotubes have a fast saturable absorption over a broad wavelength range, and the demonstrated technique is an extremely simple and inexpensive method for making fiber-integrated, saturable absorbers for passive modelocking of fiber lasers. Pulse widths of 247 fs are demonstrated from an erbium-doped fiber laser operating at 1560 nm, and 137 fs pulses are demonstrated from an amplified Yb-doped fiber laser at 1070 nm. PMID- 19547260 TI - Imaging with extended focal depth by means of lenses with radial and angular modulation. AB - The paper presents imaging properties of modified lenses with the radial and the angular modulation. We analyze three following optical elements with moderate numerical apertures: the forward logarithmic axicon and the axilens representing the radial modulation as well as the light sword optical element being a counterpart of the axilens with the angular modulation. The abilities of the elements for imaging with extended depth of focus are discussed in detail with the help of structures of output images and modulation transfer functions corresponding to them. According to the obtained results only the angular modulation of the lens makes possible to maintain the acceptable resolution, contrast and brightness of the output images for a wide range of defocusing. Therefore optical elements with angular modulations and moderate numerical apertures seem to be especially suitable for imaging with extended focal depth. PMID- 19547261 TI - Electrically reconfigurable silicon microring resonator-based filter with waveguide-coupled feedback. AB - We demonstrate an electrically reconfigurable silicon microring resonator-based filter with waveguide-coupled feedback. Our experiments and scattering-matrix based modeling show that the resonance wavelengths, extinction ratios, and line shapes depend on the feedback coupling and can be controllably tuned by means of carrier injection to the feedback- waveguide. We also demonstrate nearly uniform resonance line shapes over multiple free-spectral ranges by nearly phase-matching the feedback and the microring. PMID- 19547262 TI - Ultrafast all-optical chalcogenide glass photonic circuits. AB -

Focus Serial: Frontiers of Nonlinear Optics

Chalcogenide glasses offer large ultrafast third order nonlinearities, low two-photon absorption and the absence of free carrier absorption in a photosensitive medium. This unique combination of properties is nearly ideal for all-optical signal processing devices. In this paper we review the key properties of these materials, outline progress in the field and focus on several recent highlights: high quality gratings, signal regeneration, pulse compression and wavelength conversion. PMID- 19547263 TI - An alternative scattering method to characterize surface roughness from transparent substrates. AB - An alternative scattering method is developed to characterize surface roughness from the two faces of transparent substrates. Specific weights are attributed to each surface in the scattering process, due to the large substrate thickness. The resulting roughness spectra are shown to quasi-overlap those of near field microscopy. PMID- 19547264 TI - Thermal lens study of energy transfer in Yb(3+)/Tm(3+)-co-doped glasses. AB - Energy transfer (ET) and heat generation processes in Yb(3+)/Tm(3+)-co-doped low silica calcium-aluminosilicate glasses were investigated using thermal lens and photoluminescence measurements. Stepwise ET processes from Yb(3+) to Tm(3+), with excitation at 0.976 mum, produced efficient emission in the mid-infrared range at around 1.8 mum, with high fluorescence quantum efficiency (~0.50) and relatively low thermal loading (2) measurement states to mitigate the noise amplification effects due to singularities in the reconstruction formula. Computer-simulation studies are carried out to quantitatively and systematically investigate the developed method, within the context of propagation-based X-ray phase-contrast imaging. The reconstructed images are shown to possess dramatically reduced noise levels and greatly enhanced imaging contrast. PMID- 19547352 TI - LDPC-coded MIMO optical communication over the atmospheric turbulence channel using Q-ary pulse-position modulation. AB - We describe a coded power-efficient transmission scheme based on repetition MIMO principle suitable for communication over the atmospheric turbulence channel, and determine its channel capacity. The proposed scheme employs the Q-ary pulse position modulation. We further study how to approach the channel capacity limits using low-density parity-check (LDPC) codes. Component LDPC codes are designed using the concept of pairwise-balanced designs. Contrary to the several recent publications, bit-error rates and channel capacities are reported assuming non ideal photodetection. The atmospheric turbulence channel is modeled using the Gamma-Gamma distribution function due to Al-Habash et al. Excellent bit-error rate performance improvement, over uncoded case, is found. PMID- 19547353 TI - Optical switches based on partial band gap and anomalous refraction in photonic crystals modulated by liquid crystals. AB - Optical switches using two transmission properties in triangular photonic crystals infiltrated with liquid crystals (LCs) are investigated for incorporation in wave-guided structures for planar lightwave circuits. The two devices employ partial band gap and anomalous refraction, which are based on the anisotropic characteristics of LC reorientation under applied fields. These switches have been designed and their parameters have been analyzed by the plane wave and finite-difference time-domain calculations. In the on/off switching system, the partial band gap can be controlled when the normalized operation frequency is 0.27. The anomalous refraction can be modulated to deflect a light beam with a maximum deflection angle ~57 degrees when the frequency is 0.3. The tunability induced by LCs can create a sharp switching in the photonic devices. PMID- 19547354 TI - Surface solitons in waveguide arrays: Analytical solutions. AB - A novel phase-space method is employed for the construction of analytical stationary solitary waves located at the interface between a periodic nonlinear lattice of the Kronig-Penney type and a linear or nonlinear homogeneous medium as well as at the interface between two dissimilar nonlinear lattices. The method provides physical insight and understanding of the shape of the solitary wave profile and results to generic classes of localized solutions having either zero or nonzero semi-infinite backgrounds. For all cases, the method provides conditions involving the values of the propagation constant of the stationary solutions, the linear refractive index and the dimensions of each part in order to assure existence of solutions with specific profile characteristics. The evolution of the analytical solutions under propagation is investigated for cases of realistic configurations and interesting features are presented such as their remarkable robustness which could facilitate their experimental observation. PMID- 19547355 TI - Room temperature lasing of InAs/GaAs quantum dots in the whispering gallery modes of a silica microsphere. AB - We have achieved low threshold lasing of self-assembled InAs/GaAs quantum dots coupled to the evanescent wave of the high-Q whispering gallery modes of a silica microsphere. In spite of high temperature and Q-spoiling of whispering gallery modes due to diffusion and refraction on the high index semiconductor sample, room temperature lasing is obtained with less than 100 quantum dots. This result highlights the feasibility and interest of combining self-assembled quantum dots and microspheres in view of cavity-quantum electrodynamics experiments. PMID- 19547356 TI - A new robust regime for a dispersion-managed multichannel 2R regenerator. AB - We study the performance of a multichannel version [M. Vasilyev and T.I. Lakoba, Opt. Lett. 30, 1458 (2005)] of the all-optical Mamyshev regenerator in a practically important situation where one of its key components - a periodic group-delay device - has a realistic amplitude characteristic of a bandpass filter. We show that in this case, the regenerator can no longer operate in the regime reported in our original paper. Instead, we have found a new regime in which the regenerator's performance is robust not only to such filtering, but also to considerable variations of regenerator parameters. In this regime, the average dispersion of the regenerator must be (relatively) large and anomalous, in constrast to what was considered in all earlier studies of such (single channel) regenerators based on spectral broadening followed by off-center filtering. In addition, hardware implementation of a regenerator in the new regime is somewhat simpler than that in the original regime. PMID- 19547357 TI - Nonlinear pulse propagation in optical fibers using second order moments. AB - We present simple yet efficient formulae for the propagation of the second order moments of a pulse in a nonlinear and dispersive optical fiber over many dispersion and nonlinear lengths. The propagation of the temporal and spectral widths, chirp and power of pulses are very precisely approximated and quickly calculated in both dispersion regimes as long as the pulses are not high order solitons. PMID- 19547358 TI - Silicon-on-insulator ultra-compact duplexer based on a diffractive grating structure. AB - The use of a silicon-on-insulator diffractive grating structure is proposed to achieve ultra-compact duplexing operation. One-dimensional grating structures are proposed to spatially separate two wavelength bands. This device can become a key component in the fabrication of integrated optical transceivers for fiber-to-the home applications, where a 1310nm wavelength channel and a 1490nm wavelength channel need to be duplexed. A 10mum x 10mum one-dimensional grating structure allows to spatially separate both wavelength bands on the photonic integrated circuit, with an average coupling efficiency of 55% and an optical bandwidth of 55-60nm. While these one-dimensional grating structures are strongly polarization dependent, a two-dimensional grating structure is presented to achieve polarization independent operation. PMID- 19547359 TI - Local-field enhancement and plasmon tuning in bimetallic nanoplanets. AB - A full-interaction electromagnetic approach is applied to interpret the local- and far-field properties of AuAg alloy nanoplanets (i.e. a central cluster surrounded by small "satellite" clusters very close to its surface) fabricated in silica by ion implantation and ion irradiation techniques. Optical extinction spectroscopy reveals a large plasmon redshift which is dependent on the irradiation conditions. Simulations strongly suggest that the peculiar topological arrangement of the satellite clusters is responsible for the observed plasmonic features. Theoretical results also indicate that strong local-field enhancement is obtained between coupled clusters. Calculations for Ag models show that enhancement factors as high as ~100 are readily achievable. PMID- 19547360 TI - SNR enhancement through phase dependent signal reconstruction algorithms for phase separated interferometric signals. AB - We report several signal reconstruction algorithms for processing phase separated homodyne interferometric signals. Methods that take advantage of the phase of the signal are experimentally shown to achieve a signal-to-noise ratio (SNR) improvement of up to 5 dB over commonly used algorithms. To begin, we present a derivation of the SNR resulting from five image reconstruction algorithms in the context of a 3x3 fiber-coupler based homodyne optical coherence tomography (OCT) system, and clearly show the improvement in SNR associated with phase-based algorithms. Finally, we experimentally verify this improvement and demonstrate the enhancement in contrast and improved image quality afforded by these algorithms through homodyne OCT imaging of a Xenopus laevis tadpole. These algorithms can be generally applied in signal extraction processing where multiple phase separated measurements are available. PMID- 19547361 TI - Differential imaging in coherent anti-Stokes Raman scattering microscopy with Laguerre- Gaussian excitation beams. AB - We propose a new differential imaging technique to visualize the fine structures and the edges of a sample in coherent anti-Stokes Raman Scattering (CARS) microscopy. Both the pump and Stokes excitation fields are modulated simultaneously with a spiral phase mask which transforms them from Gaussian modes into Laguerre-Gaussian modes of LG01 for CARS excitation. With an accurate three dimensional finite-difference time-domain (FDTD) method, the intensity and phase distributions of focused input fields, the scattering pattern of generated CARS signal as well as the formation of differential images are studied detailedly, and by simulating the sensitivity range and reliability of this method, we have verified that it is much suitable for visualizing structures with a scale comparable to the excitation wavelength and has higher reliability in retrieving chemical structural information of the sample compared to common CARS microscopy. PMID- 19547363 TI - Refractive index modification using fs-laser double pulses. AB - Buried waveguides in glass are manufactured by irradiation with femtosecond laser double pulses. The refractive index change Deltan is determined by measuring the numerical aperture NA of the waveguides and by through light microscopy. The value of Deltan shows a significant dependency on the time delay Deltat of the fs laser double pulses. A Deltan of up to 2x10(-3) in fused silica is reached at a Deltat between 400 and 800 ps. Based on the results of the double pulse experiments the initial effects of the refractive index change are discussed, taking into account thermal effects and the formation of self trapped excitons (STE) and transient color centers and their interaction with the next laser pulse. PMID- 19547362 TI - Multidimensional non-linear laser imaging of Basal Cell Carcinoma. AB - We have used a multidimensional non-linear laser imaging approach to visualize ex vivo samples of basal cell carcinoma (BCC). A combination of several non-linear laser imaging techniques involving fluorescence lifetime, multispectral two photon and second-harmonic generation imaging has been used to image different skin layers. This approach has elucidated some morphological (supported by histopathological images), biochemical, and physiochemical differences of the healthy samples with respect to BCC ones. In particular, in comparison with normal skin, BCC showed a blue-shifted fluorescence emission, a higher fluorescence response at 800 nm excitation wavelength and a slightly longer mean fluorescence lifetime. Finally, the use of aminolevulinic acid as a contrast agent has been demonstrated to increase the constrast in tumor border detection. The results obtained provide further support for in-vivo non-invasive imaging of Basal Cell Carcinoma. PMID- 19547364 TI - Coherent femtosecond pulse shaping for the optimization of a non-linear micro endoscope. AB - A flexible multicore fiber bundle is fed by temporally and spectrally shaped femtosecond pulses allowing for the pre-compensation of both chromatic dispersion and non-linear effects encountered in the bundle. We demonstrate that the pulse duration at the fiber bundle output can be significantly reduced in comparison with linear pre-compensation only. The scheme for femtosecond pulse fiber delivery is applied to the optimization of two-photon fluorescence (TPF) imaging. Experiments and calculations show a five-fold improvement of the TPF signal produced at the end of the fiber bundle in comparison with linear pre compensation. This is applied to the recording, in real time (12 image/s), of TPF laser-scanning images of human colon cells stained with a fluorescent marker. Further optimizations are discussed. PMID- 19547365 TI - Offset-apertured near-field scanning optical microscope probes. AB - Near-field scanning optical microscope (NSOM) probe designs consisting of a subwavelength aperture offset of either a metallic or metal-coated dielectric cantilevered tip are investigated using finite-difference time-domain calculations. The offset aperture and metal-coated dielectric tip couple surface plasmons that illuminate the tip apex, which results in a single-lobed probing optical spot having a full-width half maximum (FWHM) similar to the apex diameter. Since the surface plasmons converge at the apex, an offset-apertured probe promises significantly higher throughput light intensities than an apertured NSOM having a comparable spot FWHM. PMID- 19547366 TI - Optical characterization of polymer liquid crystal cell exhibiting polymer blue phases. AB - The optical properties of polymer liquid crystal cell exhibiting polymer blue phases (PBPs) have been determined using ultraviolet-visible spectrophotometry, polarizing optical microscopy (POM), differential scanning calorimetry (DSC), X ray measurements, FTIR imaging and optical rotation technique. PBPs are thermodynamically stabile mesophases, which appear in chiral systems between isotropic and liquid crystal phases. A series of cyclosiloxane-based blue phase polymers were synthesized using a cholesteric LC monomer and a nematic LC monomer, and some of the polymers exhibit PBPs in temperature range over 300 degrees in cooling cycles. The unique property based on their structure and different twists formed and expect to open up new photonic application and enrich polymer blue phase contents and theory. PMID- 19547367 TI - Two-photon entanglement generation: different Bell states within the linewidth of phase-matching. AB - It is shown that for a phase-matched nonlinear process producing entangled states, different Bell states are generated for different mismatch values. In particular, generation of the singlet Bell state is demonstrated within the natural linewidth of collinear frequency-degenerate type-II spontaneous parametric down-conversion (SPDC) without the o-e delay compensation. The singlet state can be filtered out by spectral selection or by the time selection of the two-photon amplitude at the output of a dispersive fibre. The effect is of considerable importance for fibre quantum communication. PMID- 19547368 TI - Storage capacity of slow-light tunable optical buffers based on fiber Brillouin amplifiers for real signal bit streams. AB - We present a theoretical analysis of the slow-light tunable optical buffers based on fiber Brillouin amplifiers (FBA). Its storage capacity was discussed for return-to-zero (RZ) and non-return-to-zero (NRZ) bit streams. Gain saturation and pulse broadening are two key factors which limit the buffer capacity. Gain saturation is an inherent characteristic of the FBA. Broadening of the amplified pulse always accumulates with increasing gain owing to the dispersion during stimulated Brillouin scattering (SBS) process. It is shown that the maximum buffer capacity varies with data bit rate and for continuous wave (CW) or quasi CW pump it is 0.53 and 1.04 bit for RZ and NRZ respectively. Also, the optimum data bit rate to achieve the best storage capacity is obtained. PMID- 19547369 TI - Apodized coupled resonator waveguides. AB - In this paper we propose analyse the apodisation or windowing of the coupling coefficients in the unit cells of coupled resonator waveguide devices (CROWs) as a means to reduce the level of secondary sidelobes in the bandpass characteristic of their transfer functions. This technique is regularly employed in the design of digital filters and has been applied as well in the design of other photonic devices such as corrugated waveguide filters and fiber Bragg gratings. The apodisation of both Type-I and Type-II structures is discussed for several windowing functions. PMID- 19547370 TI - Soliton dynamics and self-induced transparency in nonlinear nanosuspensions. AB - We study spatial soliton dynamics in nano-particle suspensions. Starting from the Nernst-Planck and Smoluchowski equations, we demonstrate that in these systems the underlying nonlinearities as well as the nonlinear Rayleigh losses depend exponentially on optical intensity. Two different nonlinear regimes are identified depending on the refractive index contrast of the nanoparticles involved and the interesting prospect of self-induced transparency is demonstrated. Soliton stability is systematically analyzed for both 1D and 2D configurations and their propagation dynamics in the presence of Rayleigh losses is examined. The possibility of synthesizing artificial nonlinearities using mixtures of nanosuspensions is also considered. PMID- 19547371 TI - Directly measuring the spatio-temporal electric field of focusing ultrashort pulses. AB - We present the first technique for directly measuring (without assumptions) the spatio-temporal intensity and phase of a train of ultrashort pulses at and near a focus. Our method uses an experimentally simple and high-spectral resolution variant of spectral interferometry (SEA TADPOLE). To illustrate our technique, we measured the spatio-temporal electric field in and around the foci of several different types of lenses. To confirm our results, we also simulated these measurements by numerically propagating a pulse through each of the lenses used. From one set of measurements, we made a movie showing a focusing pulse with severe chromatic aberration. PMID- 19547372 TI - Boundary integral method for the challenging problems in bandgap guiding, plasmonics and sensing. AB - A boundary integral method [1] for calculating leaky and guided modes of microstructured optical fibers is presented. The method is rapidly converging and can handle a large number of inclusions (hundreds) of arbitrary geometries. Both, solid and hollow core photonic crystal fibers can be treated efficiently. We demonstrate that for large systems featuring closely spaced inclusions the computational intensity of the boundary integral method is significantly smaller than that of the multipole method. This is of particular importance in the case of hollow core band gap guiding fibers. We demonstrate versatility of the method by applying it to several challenging problems. PMID- 19547373 TI - Increasing upstream capacity in TDM-PON with multiple-wavelength transmission using Fabry-Perot laser diodes. AB - We propose a new technique for multiple-wavelength upstream transmission in time division multiplexed-passive optical networks using Fabry-Perot laser diodes (FP LD) at optical network units (ONU). The FP-LD transmits at one of strategically separated seeding wavelengths from the optical line terminal enabling the ONUs to join one of few TDM upstream channels. The scheme increases upstream capacity without the use of costly, higher speed burst mode transceivers. We present experimental results showing that up to 9 upstream channels at 2.5 Gb/s data rate can be achieved with this scheme. The paper presents locking characteristics of the FP-LD relevant for this application such as suppression of other seeding wavelengths, minimum wavelength separation and burst mode operation. PMID- 19547374 TI - Resistance of the double random phase encryption against various attacks. AB - Several attacks are proposed against the double random phase encryption scheme. These attacks are demonstrated on computer-generated ciphered images. The scheme is shown to be resistant against brute force attacks but susceptible to chosen and known plaintext attacks. In particular, we describe a technique to recover the exact keys with only two known plain images. We compare this technique to other attacks proposed in the literature. PMID- 19547375 TI - Integral imaging with large depth of field using an asymmetric phase mask. AB - We propose to improve the depth of field of Integral Imaging systems by combining an array of phase masks with the traditional lenslet array. We show that obtained elemental images are sharp over a larger range than with a regular lenslet array. We further increase the quality of elemental images by a digital restauration. Computer simulations of pickup and reconstruction are presented. PMID- 19547376 TI - Experimental observation of slow light in photonic crystal coupled waveguides. AB - We experimentally demonstrate wideband dispersion-free slow light in chirped photonic crystal coupled waveguides (PCCW). In unchirped PCCWs, the zero group velocity can occur at an inflection point of a photonic band of even symmetric mode. The even symmetric mode is selectively excited by connecting the device with input and output waveguides through optimized branch and confluence structures. In the device fabricated on SOI substrate, a large increase in group delay was observed with a maximum group index of 140 and the zero group velocity dispersion at the inflection point. Photonic bands estimated from the group delay characteristics corresponded to calculated ones. In the chirped PCCWs, the group velocity dispersion was internally compensated and the nearly constant group index of 50-60 was obtained in a wavelength bandwidth of 10 nm. The dispersion compensation was also confirmed through the transmission measurement of sub-ps optical pulses. PMID- 19547377 TI - Observation of red-shifted strong surface plasmon scattering in single Cu nanowires. AB - Surface plasmon scattering spectra of chemically produced single Cu nanowires were obtained using a total internal reflection microscope. In particular, we have observed a strong surface plasmon peak in the far red and a red-shift of the surface plasmon resonance with increasing nanowire diameter. We believe that the most reasonable origin for the red-shift of comparably large diameter nanowires is the phase retardation effect. PMID- 19547378 TI - Correlated photon-pair generation in reverse-proton-exchange PPLN waveguides with integrated mode demultiplexer at 10 GHz clock. AB - We report 10-ps correlated photon pair generation in periodically-poled reverse proton-exchange lithium niobate waveguides with integrated mode demultiplexer at a wavelength of 1.5-mum and a clock of 10 GHz. Using superconducting single photon detectors, we observed a coincidence to accidental count ratio (CAR) as high as 4000. The developed photon-pair source may find broad application in quantum information systems as well as quantum entanglement experiments. PMID- 19547379 TI - Lasing dynamics of a silicon photonic crystal microcavity. AB - In this paper we propose a novel silicon microcavity design based on the dispersion engineered photonic crystals (PhCs). With the unique self-collimation property of PhCs, we optimize the passive cavity by tuning the design parameters, such as coupling gap size and array size, to achieve higher Q factor and drop efficiency. Highest cavity mode below the band edge is of particular interest. The strong mode confinement in the low index active material offers an opportunity to realize a lasing mechanism. To investigate the lasing dynamics we introduce the rate equations of atomic system into the electromagnetic polarization to fully describe the nonlinearity of active medium. With these auxiliary differential equations we solve the time evolutions of the electromagnetic waves and atomic populations by using the FDTD method. PMID- 19547380 TI - Computational model for nonlinear plasma formation in high NA micromachining of transparent materials and biological cells. AB - Cell surgery based on ultrashort laser pulses is a fast evolving field in biophotonics. Noninvasive intra cellular dissection at sub-diffraction resolution can be performed within vital cells with very little hazardous effects to adjacent cell organelles. Microscope objectives of high numerical aperture (NA) are used to focus ultrashort pulses to a small spot. Due to the high order of nonlinearity, plasma formation and thus material manipulation is limited to the very focus. Nonetheless nonlinear plasma formation is generally accompanied by a number of additional nonlinear effects like self-focusing and filamentation. These parasitic effects limit the achievable precision and reproducibility of applications. Experimentally it is known that the intensity of these effects decreases with increasing NA of the focusing optics, but the process of nonlinear plasma formation at high NA has not been studied numerically in detail yet. To simulate the interaction of ultrashort laser pulses with transparent materials at high NA a novel nonlinear Schr odinger equation is derived; the multiple rate equation (MRE) model is used to simultaneously calculate the generation of free electrons. Nonparaxial and vectorial effects are taken into account to accurately include tight focusing conditions. Parasitic effects are shown to get stronger and increasingly distortive for NA < 0.9, using water as a model substance for biological soft tissue and cellular constituents. PMID- 19547381 TI - Collapse arrest and self-guiding of femtosecond pulses. AB - Nonlinear evolution of femtosecond pulses in media with weak dispersion and power slightly above the critical for self-focusing in the framework of generalized non paraxial amplitude equation is analyzed. It is found that this nonlinear non paraxial regime strongly depends from the initial form of the pulses. In case of long pulse (small transverse and large longitudinal size), the dynamics is closer to nonlinear paraxial dynamics of a laser beam, and the difference consists in large spectral and longitudinal spatial modulation of the long pulse. The non paraxial terms play an important role on the evolution of light bullets and light disks. In regime of light bullets (relatively equal transverse and longitudinal size) weak self-focusing without pedestal and collapse arrest is obtained. Non collapsed regime of light disks (pulses with small longitudinal and large transverse size) is also observed. Our results are in good agreement with the recent experiments on nonlinear propagation of femtosecond pulses. For first time is demonstrated that such non-paraxial model can explain effects as spectral broadening, collapse arrest and nonlinear wave guide behavior. PMID- 19547382 TI - Enhanced Kerr nonlinearity in sub-wavelength diameter As(2)Se(3) chalcogenide fiber tapers. AB - We experimentally demonstrate enhanced Kerr nonlinear effects in highly nonlinear As(2)Se(3) chalcogenide fiber tapered down to sub-wavelength waist diameter of 1.2 mum. Based on self phase modulation measurements, we infer an enhanced nonlinearity of 68 W(-1)m(-1). This is 62,000 times larger than in standard silica singlemode fiber, owing to the 500 times larger n(2) and almost 125 times smaller effective mode area. We also consider the potential to exploit the modified dispersion in these tapers for ultra-low threshold supercontinuum generation. PMID- 19547383 TI - DBR, Sub-wavelength grating, and Photonic crystal slab Fabry-Perot cavity design using phase analysis by FDTD. AB - We demonstrate a Finite-Difference Time-Domain (FDTD) phase methodology to estimate resonant wavelengths in Fabry-Perot (FP) cavity structures. We validate the phase method in a conventional Vertical-Cavity Surface-Emitting Laser (VCSEL) structure using a transfer-matrix method, and compare results with a FDTD reflectance method. We extend this approach to a Sub-Wavelength Grating (SWG) and a Photonic Crystal (Phc) slab, either of which may replace one of the Distributed Bragg Reflectors (DBRs) in the VCSEL, and predict resonant conditions with varying lithographic parameters. Finally, we compare the resonant tunabilities of three different VCSEL structures, taking quality factors into account. PMID- 19547384 TI - Birefringent in-phase supermode operation of a multicore microstructured fiber laser. AB - We report the first observation of birefringent in-phase supermode operation of a phase-locked multicore fiber laser. The in-phase mode operation of our 12-core rectangular-array microstructured fiber laser was confirmed by the near-field distribution, the far-field diffraction pattern, and the optical spectrum. The birefringence of the in-phase mode in propagation constant Deltay was measured as ~ 4 x 10(-6) 1/mum. The break of the polarization degeneracy indicates the possibility of single polarization operation of phase-locked multicore fiber lasers and amplifiers. PMID- 19547385 TI - Densely integrated microring resonator based photonic devices for use in access networks. AB - Two reconfigurable optical add-drop multiplexers, operating in the second or third telecom window, as well as a 1x4x4 reconfigurable lambda-router operating in the second telecom window, are demonstrated. The devices have a footprint less than 2 mm(2) and are based on thermally tunable vertically coupled microring resonators fabricated in Si(3)N(4)/SiO(2). PMID- 19547386 TI - Efficient optical modeling of spontaneous emission in a cylindrically layered nanostructure. AB - We present an efficient optical model to study spontaneous emission in a cylindrically layered nanostructure. The total emission power of an emitter in the nanostructure is efficiently calculated. A formula is derived to calculate the lateral-surface emission power. As examples of practical interest, spontaneous emission properties, including radiative transition rate of the emitter, the assignment of the emission to lateral-surface emission and waveguided emission, are comprehensively studied at the first time for an isolated ZnO nanowire and a ZnO/SiO(2) nanocable. PMID- 19547387 TI - Coupled resonator optical waveguide structures with highly dispersive media. AB - Analysis of photonic crystal coupled resonator optical waveguide (CROW) structures with a highly dispersive background medium is presented. A finite difference time-domain algorithm was employed which contains an exact representation of the permittivity of a three-level atomic system which exhibits electromagnetically induced transparency (EIT). We find that the coupling strength between nearestneighbor cavities in the CROW decreases with increasing steepness of the background dispersion, which is continuously tunable as it is directly related to the control field Rabi frequency. The weaker coupling decreases the speed of pulse propagation through the waveguide. In addition, due to the dispersive nature of the EIT background, the CROW band shape is tuned around a fixed k-point. Thus, the EIT background enables dynamic tunability of the CROW band shape and the group velocity in the structure at a fixed operating point in momentum space. PMID- 19547388 TI - Ultra-sensitive wavefront measurement using a Hartmann sensor. AB - We describe a Hartmann sensor with a sensitivity of lambda /15,500 at lambda= 820nm. We also demonstrate its application to the measurement of an ultra small change in wavefront and show that the result agrees with that expected to within lambda/3,300. PMID- 19547389 TI - Emission spectrum broadening of Nd:YVO(4) with femtosecond laser pulse processing. AB - The observation of emission spectrum broadening of Nd:YVO(4) by femtosecond laser pulse processing is reported. This result opens the possible way of sub-ps pulse generation from Nd ion doped crystals with better physical qualities than that of glass based laser gain materials. PMID- 19547390 TI - Characterization of a nematic PALC at large oblique incidence angles. AB - Compared with conventional photometric methods of measuring cell parameters, including the cell gap and the pretilt angle of a nematic parallel-aligned liquid crystal (PALC) using multiple wavelengths at normal incidence, this research proposes the use of a phase-sensitive interferometric ellipsometer to determine cell parameters precisely based on a single wavelength at large oblique incidence angles. The advantage of this method is that it detects the phase difference using an optical heterodyne interferometer in which a common phase noise rejection mode is provided. Thus, there is a high signal-to-noise ratio (SNR) on the phase measurement. In addition, a range of large oblique incidence angles on the PALC is used so that a high sensitivity measurement of the cell parameters is obtained experimentally. During the measurements, the multiple reflections and spatial shifting effect of the emerging extraordinary ray (E-ray) and ordinary ray (O-ray) from the PALC at large oblique incidence angles are able to be reduced effectively by the use of retro-reflected geometry in the interferometer. The experimental results verify that the sensitivities for the cell gap and pretilt angle measurements are 0.3 nm and 0.01 degrees , respectively. PMID- 19547391 TI - In vivo three-dimensional microelectromechanical endoscopic swept source optical coherence tomography. AB - In clinical applications, three-dimensional (3-D) microscopic image volume reveals tissue morphological changes, which are closely related to pre-cancer and early stage disease, much better than two-dimensional images. However, the traditional endoscope only achieves two-dimensional surface images. In this paper, a 3-D endoscopic microscope was developed based on a rotational microelectromechanical system (MEMS) probe [1]. The 3-D helix scan mode was realized by combining a MEMS motor rotational scan and linear stage transversal movement. In order to coordinate the high spin speed of MEMS motor inside the endoscope, an optical coherence tomography (OCT) system with a broadband fast swept laser was used. In vivo 3-D image volumes of rabbit esophagus and trachea were demonstrated. PMID- 19547392 TI - Stimulated Brillouin scattering modeling for high-resolution, time-domain distributed sensing. AB - Starting from the standard three-wave SBS coupled equations, we derive a novel expression describing Brillouin interaction between a pulsed pump wave with a finite cw component, and a Stokes continuous wave counter-propagating along a single-mode optical fiber. The derived integral equation relates the time-domain Stokes beam amplification to the Brillouin frequency distribution. The proposed model permits an accurate description of the Brillouin interaction even for arbitrarily-shaped pump pulses, and can be efficiently employed for improving the accuracy and the resolution of SBS-based distributed sensors. The validity and the limits of the proposed model are numerically analyzed and discussed. PMID- 19547393 TI - Coherent anti-Stokes Raman scattering (CARS) microscopy imaging at interfaces: evidence of interference effects. AB - We show in this paper that the contrast of the interface between resonant and nonresonant media imaged in Coherent anti-Stokes Raman scattering (CARS) microscopy strongly depends on the pump and Stokes fields spectral detuning. More specifically, when this detuning drives the vibrational resonance with the maximum phase difference, a spatial dip appears at the interface in the CARS image. This effect is studied both theoretically and experimentally and is an evidence of the coherent and resonant nature of the CARS contrast mechanism. PMID- 19547394 TI - Optical imaging for medical diagnosis based on active stereo vision and motion tracking. AB - This study aims to develop a novel imaging technique to improve the accuracy of the colposcolpic diagnosis of cervical cancer. An optical imaging system based on active stereo vision is built to measure the 3-D surface topology of cervix and track the motion of patient. The information of motion tracking are used to register the time-sequenced images of cervix recorded over the period of examination. The imaging system is evaluated by tracking the movements of cervix models. The results show that the error of 2-D image registration is 0.8 pixels, equivalent to the motion tracking error of 0.05 mm in the field-of-view. The imaging technique holds the promise to enable quantitative mapping of the acetowhitening kinetics over cervical surface for more accurate diagnosis of cervical cancer. PMID- 19547395 TI - High transmission through ridge nano-apertures on Vertical-Cavity Surface Emitting Lasers. AB - We report high-intensity nano-aperture Vertical-Cavity Surface- Emitting Lasers (VCSELs) with sub-100nm near-field spots using ridge apertures. Power transmission efficiency through different ridge apertures, including bowtie, C, H and I-shaped apertures on VCSELs were studied. Significantly higher transmission efficiencies were obtained from the ridge apertures than those from conventional square apertures. Mechanisms for high transmission through the ridge apertures are explained through simulation and waveguide theory. A new quadruple-ridge aperture is proposed and designed via simulation. With the high-intensity and small spot size, VCSELs using these ridge nano-apertures are very promising means to realize applications such as ultrahigh-density near-field optical data storage and ultrahigh-resolution near-field imaging etc. PMID- 19547396 TI - Whole spectrum fluorescence detection with ultrafast white light excitation. AB - We have developed a new detection mechanism for ultrabroadband multicolor fluorescence detection using an ultrafast supercontinuum white light source without spectral filtering to simultaneously excite different fluorophores. A nonlinear photonic crystal fiber was utilized in conjunction with a femtosecond laser to generate the supercontinuum. A time-resolved detector was tested to detect the whole spectrum fluorescence while gating out the excitation white light in the time domain. PMID- 19547397 TI - Symmetry breaking of discrete solitons and its suppression by partial incoherence. AB - We study the symmetry breaking instability of discrete solitons with even parity in a 1-D waveguide array, and find that such instability can be suppressed by adding spatial incoherence. This is true for both staggered and unstaggered modes. PMID- 19547398 TI - Encapsulation shape with non-rotational symmetry designed for extraction of polarized light from unpolarized sources. AB - A non-rotationally symmetric encapsulation shape - which takes advantage of the low reflection coefficient for transverse magnetic polarized light near Brewster's angle - designed to enhance extraction of a particular desired linear polarization from an unpolarized source is reported. The algorithm for optimization of the shape is described. Numerical ray-tracing simulations of the encapsulation shape are performed and predict an integrated enhancement of 8.3% in the ratio of desired polarization to undesired polarization when the refractive index of the encapsulant is 1.5. Experimental measurements of fabricated encapsulant shapes agree well with numerical predictions. PMID- 19547399 TI - Assessment of the horizontal resolution of retrieval products derived from MIPAS observations. AB - We report the results of a study aimed at the assessment of the trade-off between precision and horizontal resolution of the retrieval products of MIPAS (Michelson Interferometer for Passive Atmospheric Sounding) operating onboard the ENVIronmental SATellite. By exploiting different observation setups we could perform the study by acting on both the retrieval and the sampling grids. Our results are compared with those previously obtained on simulated observations [Appl. Opt. 43, 1-11 (2004)]. We show that the horizontal sampling of the atmosphere operated by the spectrometer cannot be pushed beyond some limits without inducing unacceptable correlations among the retrieved profiles. These correlations show-up only when using a two-dimensional retrieval algorithm and can be evaluated through the instabilities that they trigger in the horizontal distribution of the retrieval products. In order to reduce these instabilities we compare the strategy of degrading the retrieval grid with the strategy of applying horizontal regularization. We discuss the different trade-off between precision and spatial resolution connected with the two strategies. The method adopted in this study, is applicable to any orbiting limb sounder measuring along the orbit track. PMID- 19547400 TI - An optical solution for the traveling salesman problem. AB - We introduce an optical method based on white light interferometry in order to solve the well-known NP-complete traveling salesman problem. To our knowledge it is the first time that a method for the reduction of non-polynomial time to quadratic time has been proposed. We will show that this achievement is limited by the number of available photons for solving the problem. It will turn out that this number of photons is proportional to N(N) for a traveling salesman problem with N cities and that for large numbers of cities the method in practice therefore is limited by the signal-to-noise ratio. The proposed method is meant purely as a gedankenexperiment. PMID- 19547401 TI - Homogenized LED-illumination using microlens arrays for a pocket-sized projector. AB - We present an LED-based ultra-mini DMD projector with a size of 75 mm x 67 mm x 42 mm. A compact homogenizer consisting of a double-side microlens array and two condensers was proposed to reduce the size of the pocket-sized projector. The homogenizer not only allowed for a reduction in the total track length of the system, but also reduced the angle of the rays emitted from the LED with the micro field lens array. The double-side well-aligned 124 x 146 microlens array was fabricated using backside alignment and hot embossing techniques. The microlens array was square-arranged and the fill-factor was extremely high. The uniformity and total throughput of this projector were higher than those of the current pocket-sized projectors. Moreover, the optical performances of the projector such as color difference and the LED alignment tolerance were also measured and discussed. PMID- 19547403 TI - Double reflection in the concave reflective blazed grating. AB - The miniature spectrometer has many applications in integrated optics and photonics. The blazed grating with the Rowland circle structure has the advantage of self-focusing and is chosen as the major component in the spectrometer chip. In the simulations for the blaze angle design in the visible spectrum, we discover the phenomenon of the double reflection diffraction. Its cause and parameter space are discussed. The spectrometer utilizing the phenomenon has similar performance to the standard blazed grating and is easier to manufacture in microelectromechanical systems (MEMS) technology. The discovery will greatly ease the design of the spectrometer chip. PMID- 19547402 TI - Optical data packet synchronization and multiplexing using a tunable optical delay based on wavelength conversion and inter-channel chromatic dispersion. AB - 10 Gb/s non-return-to-zero (NRZ) on-off keyed (OOK) optical data packets are synchronized and time-multiplexed using a 26-ns tunable all-optical delay line. The delay element is based on wavelength conversion in periodically poled lithium niobate (PPLN) waveguides, inter-channel chromatic dispersion in dispersion compensating fiber (DCF) and intra-channel dispersion compensation with a chirped fiber Bragg grating (FBG). Delay reconfiguration time is measured to be less than 300 ps. PMID- 19547404 TI - Analysis of SFM dynamics in solitary and optically-injected VCSELs. AB - The aim of this paper is to use the spin-flip model (SFM) to investigate how the fundamental parameters of birefringence, spin relaxation and pumping affect the dynamics of solitary and optically-injected vertical-cavity surface-emitting lasers (VCSELs). The SFM predicts that a solitary VCSEL can exhibit diverse polarisation behaviour, including elliptical and linear stability, which can be used in polarisation switching. For given values of pumping, spin relaxation and birefringence, we analyse the electric field components, the carrier densities corresponding to spin-up and spin-down, and the relaxation oscillation frequency for a solitary VCSEL and the stability map for an optically-injected VCSEL. PMID- 19547405 TI - Narrowband DWDM filters based on Fibonacci-class quasi-periodic structures. AB - In this paper, we propose a narrowband DWDM filter structure, whose reflection band characteristics, meets the ITU-T standard. The proposed filter structure is based on Fibonacci quasi-periodic structures composed of multilayers with large index differences. Studying the effects of the optical and geometrical parameters of Fibonacci quasi-periodic structures on its filtering properties, we have realized that to achieve the ITU-T standard, we need to cascade two successive structures both with the same generation numbers j=4 and orders n=25 and apodized refractive indices. The apodization process helps to minimize the stop band sidelobes. We have also demonstrated that beside Fibonacci's order, n, the layers dimensions, and their refractive index ratios are the main design parameters. PMID- 19547406 TI - Coupling localized and extended plasmons to improve the light extraction through metal films. AB - Efficient transmission of light through a metal layer has become a key issue for a variety of applications including light-emitting diodes and solar cells. We report here on a novel strategy where localized and extended surface plasmons are combined to maximize the fluorescence transmission through a metallic film. We show that the dispersion of an artificial material formed by an array of metal nanoparticles coupled to a flat metal layer can be engineered to make the metal film, in a specific direction, 100% transmissive. PMID- 19547407 TI - Short period holographic structures for backlight display applications. AB - The use of holographic structures is promising for the realization of efficient systems in backlight applications for displays. By applying surface relief gratings on top of a side-lit lightguide we realize a backlight that avoids the use of color filters. The grating is used as a light outcoupling and color separating element. The demands for this grating are stringent and calculations have been performed to meet them. A prototype backlight, including the grating structure, has been assembled and characterized. Results of experiments are discussed. PMID- 19547408 TI - Modeling and measurement of losses in silicon-on-insulator resonators and bends. AB - We present an analytical model to quantify losses in resonators and bends without uncertain contributions from fiber coupling in/out or waveguide cleavage facets. With resonators in add-drop configuration, intrinsic losses are calculated from the free spectral range, through-port extinction and drop-port bandwidth. We fabricated and characterized silicon-on-insulator resonator for loss analysis. At 1.55 mum, racetrack resonators with a bending radius of 4.5 mum show intrinsic losses as small as 0.14+/-0.014 dB/round-trip. Meanwhile, intrinsic losses increase up to 1.23 dB/round-trip in the racetrack resonator that has a bending radius of 2.25 mum. Losses in a 180 degrees bend are estimated as a half of the intrinsic losses in these racetrack resonators, i.e., 0.07+/-0.007 dB/turn for a bending radius of 4.5 mum and 0.62 dB/turn for a bending radius of 2.25 mum. Loss in a 90 degrees bend with a radius of 4.5 mum is determined to be 0.06+/-0.006 dB/turn at 1.55 mum. The losses in 180 degrees or 90 degrees bends are found to be mainly due to the transition loss between waveguide bends and straight waveguides. PMID- 19547409 TI - 2D light scattering patterns of mitochondria in single cells. AB - The ability to characterize the mitochondria in single living cells may provide a powerful tool in clinical applications. We have recently developed a 2D (both polar angle and azimuth angle dependences) light scattering cytometric technique which we apply here to assess experimental 2D light scattering patterns from single biological cells (yeast and human). We compare these patterns to those obtained from simulations using a 3D Finite-Difference Time-Domain (FDTD) method and demonstrate that microstructure (e.g., the cytoplasm and/or nucleus) of cells generates fringes of scattered light, while in the larger human cells the light scattered by the mitochondria dominates the scatter pattern, forming compact regions of high intensity that we term 'blobs'. These blobs provide information on the mitochondria within the cell and their analysis may ultimately be useful as a diagnostic technique. PMID- 19547410 TI - Interband photorefractive effect in beta-BBO crystal due to multiphoton excitation by intense ultrashort optical pulses. AB - This paper presents the first experimental observation of interband photo- refractive (PR) effects in beta-BBO crystal due to multiphoton excitation with intense ultrashort pulses. In order to fully characterize the PR effects, a sensitive intracavity scheme is developed to magnify the dynamics of nonlinear lenses induced by the PR effects. The reproducible PR phenomena depend strongly on the power, wavelength, and spatial intensity profile of the intense laser pulses and the electro-optic coefficient of the optical materials. Its response time is from tens of seconds to several minutes. The results may be very helpful for us to find a solution to overcome the deleterious influence of multiphoton induced photo-charges on nonlinear optical frequency conversions, e.g. optical parametric chirped pulse amplification. PMID- 19547411 TI - Phase change characteristics of aluminum doped Ge(2)Sb(2)Te(5) films prepared by magnetron sputtering. AB - Aluminum-doped Ge(2)Sb(2)Te(5) (Al(x)GST) films were deposited on Si(100) substrates by co-magnetron sputtering system. The Aluminum concentrations in these films are determined by X-ray photoelectron spectroscopy (XPS). The influence of Al doping upon phase change characteristics of these Al(x)GST alloy films has been investigated by X-ray diffraction (XRD) and a temperature regulable UVISEL(TM) typed spectroscopic ellipsometry (TRSE). With the augment of Al doping concentration, the crystalline temperatures of Al(x)GST films went up while annealing, and the face-centered-cubic (fcc) phase had high thermal stability. The reflectivity contrast of the films increases obviously, which is effective to improve the signal to noise ratio (SNR) for optical phase-change storage. PMID- 19547412 TI - Transfer function of radio over fiber multimode fiber optic links considering third-order dispersion. AB - Although a considerable number of multimode fiber (MMF) links operate in a wavelength region around 850 nm where chromatic dispersion of a given modal group mu is described adequately by the second derivative beta(mu) (2) of the propagation constant beta(mu)(omega), there is also an increasing interest in MMF links transmitting in the second spectral window (@1300nm) where this second derivative vanishes being thus necessary to consider the third derivative beta(mu) (3) in the evaluation of the transfer function of the multimode fiber link. We present in this paper, for the first time to our knowledge, an analytical model for the transfer function of a multimode fiber (MMF) optic link taken into account the impact of third-order dispersion. The model extends the operation of a previously reported one for second-order dispersion. Our results show that the performance of broadband radio over fiber transmission through middle-reach distances can be improved by working at the minimum-dispersion wavelength as long as low-linewidth lasers are employed. PMID- 19547413 TI - A 10-Gbit/s EML link using detuned narrowband optical filtering. AB - In this paper, the effects of asymmetric narrowband optical filtering are investigated in a 10-Gbit/s optical communication link using integrated electro absorption modulated lasers (EML). We investigate the effect of EML chirp on link performance as well as the optimal filter bandwidth and wavelength detuning. We show that both the phase response and the spectral narrowing of the filter will enable a longer distance transmission by interacting with the EML transient chirp and compensating for the fiber chromatic dispersion. Experimentally, an 8.75-GHz filter is shown to improve the link distance by 40 km from 65 to 105 km, when transmitting over standard single mode fiber. PMID- 19547414 TI - Tuneable planar integrated optical systems. AB - Planar integrated free-space optical systems are well suited for a variety of applications, such as optical interconnects and security devices. Here, we demonstrate for the first time dynamic functionality of such microoptical systems by the integration of adaptive liquid-crystal-devices. PMID- 19547415 TI - Dynamic radiation force of a pulsed gaussian beam acting on rayleigh dielectric sphere. AB - We investigate the dynamic evolution of the radiation forces produced by the pulsed Gaussian beams acting on a Rayleigh dielectric sphere. We derive the analytical expressions for the scattering force and all components of the pondermotive force induced by the pulsed Gaussian beam. Our analysis shows that the radiation force can be greatly enhanced due to the effect of the short pulse duration, which leads to the enhancement of both the transverse and longitudinal radiation forces. And it is found that for the pulse with large pulse duration, it can be used for the stable trapping and manipulating the particle, while for the pulse with short pulse duration it may be used for guiding and moving the small dielectric particle. Finally we discuss the stability condition of the effectively trapping and manipulating the particle by the pulsed beam. PMID- 19547416 TI - Volume extreme ultraviolet nano-holographic imaging with numerical optical sectioning. AB - Three dimensional images were obtained using a single high numerical aperture hologram recorded in a high resolution photoresist with a table top lambda = 46.9 nm laser. Gabor holograms were numerically reconstructed over a range of image planes by sweeping the propagation distance in the numerical reconstruction algorithm, allowing numerical optical sectioning. A robust three dimension image of a test object was obtained with numerical optical sectioning, providing a longitudinal resolution of approximately 2 mum and a lateral resolution of 164 nm. PMID- 19547417 TI - Effects of bending on the performance of hole-assisted single polarization fibers. AB - We study the effects of bending on single polarization fiber performance through the use of finite element method in conjunction with the perfectly matched layer (PML) in cylindrical geometry. The cylindrical PML used in this paper allows us to calculate the loss associated with each polarization mode at a given wavelength, specified bending diameter, and specific orientation. We identified a series of bending characteristics of the single polarization fiber by choosing different bending diameters and different orientations. We also conducted experiments to study some aspects of the bending. Good qualitative agreement between numerical and experimental results is found, which helps to understand fiber deployment conditions and can potentially facilitate new design efforts. PMID- 19547418 TI - Far infrared photonic crystals operating in the Reststrahl region. AB - We report here far infrared photonic crystals comprised of a lattice-matched pair of semiconductor materials: GaP and Si, or GaAs and Ge, or AlAs and GaAs. The crystals operate in a wavelength range where the real refractive index of one material undergoes a major dispersion associated with the LO and TO phonon absorption peaks. Using electromagnetic theory, we investigated the photonic bandgap response for both TE and TM polarizations. Propagation losses for two types of crystals are estimated in this paper. These structures offer promise for the integration of III-V materials (GaP, GaAs) on group IV (Si, or Ge) for practical, active, far infrared photonic devices, such as light sources, amplifiers, modulators, reconfigurable waveguides and switches. PMID- 19547419 TI - Laser light scattering in turbid media Part I: Experimental and simulated results for the spatial intensity distribution. AB - We investigate the scattering and multiple scattering of a typical laser beam (lambda = 800 nm) in the intermediate scattering regime. The turbid media used in this work are homogeneous solutions of monodisperse polystyrene spheres in distilled water. The two-dimensional distribution of light intensity is recorded experimentally, and calculated via Monte Carlo simulation for both forward and side scattering. The contribution of each scattering order to the total detected light intensity is quantified for a range of different scattering phase functions, optical depths, and detection acceptance angles. The Lorentz-Mie scattering phase function for individual particles is varied by using different sphere diameters (D = 1 and 5 mum). The optical depth of the turbid medium is varied (OD = 2, 5, and 10) by employing different concentrations of polystyrene spheres. Detection angles of theta(a) = 1.5 degrees and 8.5 degrees are considered. A novel approach which realistically models the experimental laser source is employed in this paper, and very good agreement between the experimental and simulated results is demonstrated. The data presented here can be of use to validate any other modern Monte Carlo models which generate spatially resolved light intensity distributions. Finally, an effective correction procedure to the Beer-Lambert law is proposed based on the Monte Carlo calculation of the ballistic photon contribution to the total detected light intensity. PMID- 19547420 TI - Reflective second harmonic generation near resonance in the epitaxial Al-doped ZnO thin film. AB - The second harmonic (SH) generation from the highly epitaxial Al-doped ZnO film on sapphire was measured, using the femtosecond Ti:Sapphire laser at the near resonant SH wavelength, in reflection geometry to avoid the sapphire's contribution in the conventional Maker fringes technique. By investigating SH intensities as a function of the azimuthal angle along the film's normal, we found that the sapphire substrate had a negligible contribution to the reflective SH signal and the film had a pure and well-aligned c-domain. We also developed a new method to calculate the component's ratios of the nonlinear susceptibility tensor by analyzing the polarization diagrams of SH intensities under the incidence with two different angles. The ratios indicate that Kleinman's symmetry is broken due to the absorption at SH wavelength and the dominant component of the nonlinear susceptibility tensor is d(33). Calibration using the Z-cut quartz shows a possible overestimate of the nonlinear response by Maker fringes technique. PMID- 19547421 TI - Enhanced sensitivity digital holographic interferometry. AB - Corona discharges are generally characterized by a low optical density whose detection is often near or under the limits of interferometric techniques. In this paper, we propose a method of digital holographic interferometry that enables detection with enhanced sensitivity. This sensitivity increase is obtained by post-processing the digital holographic recordings with a set of point-wise image operations. The procedure is described mathematically and illustrated experimentally. Examples are given for an opaque object and for DC corona discharges generated in the symmetrical point-plane geometry. PMID- 19547422 TI - Phosphorescence lifetime based oxygen micro-sensing using a digital micromirror device. AB - A digital light modulation microscope (DLMM) that utilizes a digital micromirror device (DMD) on an epifluorescence microscope has been developed to modulate excitation light in spatial and temporal domains for phosphorescence lifetime detection. Local O2 concentration can be inferred through the detected lifetime around an O2-quenching phosphorescent porphyrin microsensor. Combined with microsensor arrays, the DLMM can sequentially address light to each microsensor element to construct a discrete lifetime image or O2 distribution. In contrast to conventional phosphorescence lifetime imaging, the new method eliminates the need for a pulsed light source and a time-gated camera. To demonstrate O2 sensing with lab-on-a-chip devices, an array of 150-mum-diameter micro-wells coated with phosphorescent porphyrin were observed. The locations of the sensor elements were automatically identified though image analysis. The goal of this platform is to measure the O2 consumption of individual cells trapped in the microwells. PMID- 19547423 TI - Cutoff modulator with tunable filtering characteristic based on the W type waveguide. AB - Utilizing the cutoff characteristic of zero-order TE mode of the W type waveguide, we propose a cutoff modulator with tunable filtering characteristic in this paper. Simulation results show that: for the modulator based on GaAlAs/GaAs epitaxial layers, its cutoff wavelength can be shifted from 1.62 mum to 1.42 mum when the refractive index of the core layer is decreased by -0.01 in the modulation region of 800 mum; and the extinction ration at the wavelength of 1.55 mum reaches 40 dB. Compared with other traditional cutoff modulators, it has the advantages of more compact size, simpler structure and the potential for high intensity integration. PMID- 19547424 TI - Diffractive optical elements for high gain lasers with arbitrary output beam profiles. AB - We introduce a previously unreported laser cavity configuration, using a diffractive optical element (DOE) in place of the output coupler. Such a configuration allows the DOE to work both in reflection, as a mode shaping element, and in transmission as a beam shaper. Employing dual wavelength DOE optimization techniques and phase delays greater than 2pi, allows the two functions to be designed independently. Thus, an arbitrary output beam profile can be combined with a mode shape which maximizes energy extraction from the gain medium. Devices are designed and their performance modeled for a 1m cavity with 5mm diameter mirrors and a wavelength of 632.8nm. An element with 32 quantization levels and a maximum phase delay of 8pi in transmission produces high quality results. PMID- 19547425 TI - Single-frequency Ince-Gaussian mode operations of laser-diode-pumped microchip solid-state lasers. AB - Various single-frequency Ince-Gaussian mode oscillations have been achieved in laser-diode-pumped microchip solid-state lasers, including LiNdP(4)O(12) (LNP) and Nd:GdVO(4), by adjusting the azimuthal symmetry of the short laser resonator. Ince-Gaussian modes formed by astigmatic pumping have been reproduced by numerical simulation. PMID- 19547426 TI - Spatiotemporal discrete surface solitons in binary waveguide arrays. AB - We study spatiotemporal solitons at the edge of a semi-infinite binary array of optical waveguides and, in particular, predict theoretically the existence of a novel type of surface soliton, the surface gap light bullets. We analyze the stability properties of these solitons in the framework of the continuous discrete model of an array of two types of optical waveguides. PMID- 19547427 TI - High-energy femtosecond Yb-doped dispersion compensation free fiber laser. AB - We report on a mode-locked high energy fiber laser operating in the dispersion compensation free regime. The sigma cavity is constructed with a saturable absorber mirror and short-length large-mode-area photonic crystal fiber. The laser generates positively-chirped pulses with an energy of 265 nJ at a repetition rate of 10.18 MHz in a stable and self-starting operation. The pulses are compressible down to 400 fs leading to a peak power of 500 kW. Numerical simulations accurately reflect the experimental results and reveal the mechanisms for self consistent intracavity pulse evolution. With this performance mode locked fiber lasers can compete with state-of-the-art bulk femtosecond oscillators for the first time and pulse energy scaling beyond the muJ-level appears to be feasible. PMID- 19547428 TI - Broadband fiber Bragg grating with channelized dispersion. AB - We present here a new class of multi-channel Fiber Bragg grating (FBG), which provides the characteristics of channelized dispersion but does so with only a single reflection band. An FBG of this type can provide pure phase control of the spectral waveform of optical pulses without introducing any deleterious insertion loss-variation. We anticipate that this new class of FBG will find some applications in wavelength-division- multiplexing systems. PMID- 19547429 TI - Very low voltage single drive domain inverted LiNbO(3) integrated electro-optic modulator. AB - Domain inversion is used in a simple fashion to improve significantly the performance of a waveguide electro-optic modulator in z-cut LiNbO(3). The waveguide arms of the Mach-Zehnder interferometer are placed in opposite domain oriented regions under the same, narrower and more efficient electrode, so that opposite phase shifts (push-pull effect) can still be achieved despite the arms being subjected to the same electric field. Switching voltages close to 2 V are obtained, which allow 10Gb/s modulation with inexpensive drivers, such as those used for electro-absorption modulators, which deliver driving voltages well below 3V. PMID- 19547430 TI - Refractive index and extinction coefficient dependence of thin Al and Ir films on deposition technique and thickness. AB - We show that the optical properties of thin metallic films depend on the thickness of the film as well as on the deposition technique. Several thicknesses of electron-beam-gun-evaporated aluminium films were measured and the refractive index and the extinction coefficient defined using ellipsometry. In addition, the refractive indexes and the extinction coefficients of atomic-layer-deposited iridium were compared with those of evaporated iridium samples. PMID- 19547431 TI - Preparation of high laser induced damage threshold antireflection film using interrupted ion assisted deposition. AB - Single layers and antireflection films were deposited by electron beam evaporation, ion assisted deposition and interrupted ion assisted deposition, respectively. Antireflection film of quite high laser damage threshold (18J/cm2) deposited by interrupted ion assisted deposition were got. The electric field distribution, weak absorption, and residual stress of films and their relations to damage threshold were investigated. It was shown that the laser induced damage threshold of film was the result of competition of disadvantages and advantages, and interrupted ion assisted deposition was one of the valuable methods for preparing high laser induced damage threshold films. PMID- 19547432 TI - Tracking biological microorganisms in sequence of 3D holographic microscopy images. AB - We develop a 3D region tracking method based on Maximum A Posteriori (MAP) tracker and adapt it to digital hologram sequences to efficiently track biological microorganisms in holographic microscopy data. In our approach, the target surface is modeled as the iso-surface of a level set function which is evolved at each frame via level set Hamilton Jacobian update rule in Euler Lagrangian framework. The statistical characteristics of the target microorganism versus the background are exploited to evolve the interface at each frame, thus the algorithm works independent of the shape or morphology of the target. We use the bivariate Gaussian distribution to model the reconstructed hologram data which enables us to take into account the correlation between the amplitude and phase of the reconstructed wavefront to obtain a more accurate tracking solution. PMID- 19547433 TI - Characterization of a subwavelength-scale 3D void structure using the FDTD-based confocal laser scanning microscopic image mapping technique. AB - In this paper, a simple confocal laser scanning microscopic (CLSM) image mapping technique based on the finite-difference time domain (FDTD) calculation has been proposed and evaluated for characterization of a subwavelength-scale three dimensional (3D) void structure fabricated inside polymer matrix. The FDTD simulation method adopts a focused Gaussian beam incident wave, Berenger's perfectly matched layer absorbing boundary condition, and the angular spectrum analysis method. Through the well matched simulation and experimental results of the xz-scanned 3D void structure, we first characterize the exact position and the topological shape factor of the subwavelength-scale void structure, which was fabricated by a tightly focused ultrashort pulse laser. The proposed CLSM image mapping technique based on the FDTD can be widely applied from the 3D near-field microscopic imaging, optical trapping, and evanescent wave phenomenon to the state-of-the-art bio- and nanophotonics. PMID- 19547434 TI - Investigation of symmetry of photorefractive effect in LiNbO(3). AB - The investigation of the light-induced changes of refractive index in a LiNbO(3): Fe sample in case of strip-like illumination is presented in the contribution. The changes of the refractive index are visualized by interferograms of the sample obtained for various orientation of the illuminated strip and various polarization of the light used during interferogram creation. The investigation shows that character of the dependences of the refractive index on the coordinate perpendicular to the illuminated strip is different for different strip orientation. It indicates the possibility that for different orientation of gradient of the illumination the different mechanisms are responsible for changes of the refractive index. PMID- 19547435 TI - Adaptive-mesh-based algorithm for fluorescence molecular tomography using an analytical solution: erratum. AB - An erratum is presented to correct the writing mistakes of the simulated fluorescence targets' size. PMID- 19547436 TI - Thin spectacles for myopia, presbyopia and astigmatism insensitive vision. AB - The aim of the presented research was to develop special spectacles capable of solving common ophthalmic problems as myopia, presbyopia and regular/irregular astigmatism. The method included adapting special all-optical extended depth of focus concept, taken from the field of digital imaging, to ophthalmology, and by that providing the required vision solutions. Special thin mask containing annular like replicated structure (thickness of the structure is less than one micron) was designed and proven to provide extended depth of focus. In this paper we present several experimental results as well as trials with volunteers. The testing included measuring the visual acuity under different illumination conditions (pupil size varied from 2 up to 4mm), as well as stereoscopy, color integrity, field of view and contrast. The results demonstrate improvements of up to 3 Diopters (for presbyopic that require the bifocal or the progressive lens solutions) for pupil sizes of 2-4mm. The approach has demonstrated improvement of more than 2 Diopters for regular as well as irregular astigmatism. The main advantage of the developed optical element is that it is very thin (less than few microns) and has low price, it has high energetic throughput and low chromatic aberrations and it operates over the full field of view while providing continuously focused image (in contrast to bifocal lenses having only 2 focused regions). The element also provides a solution for regular as well as irregular astigmatism that currently has no available treatment. PMID- 19547437 TI - Single-beam copying system of 360-degree viewable image-plane disk-type multiplex hologram and polarization effects on diffraction efficiency. AB - A two-step holographic process for the fabrication of an image-plane disk-type multiplex hologram is described in this paper. The diffraction efficiency of the transfer hologram is measured as a function of exposure. It is found to be influenced by the polarization of the light beams of the copying system, resulting in different diffraction efficiencies from different areas (under different interference conditions) of the hologram transfer. The factors which cause the phenomenon of diffraction-efficiency difference are demonstrated and the corresponding experimental results are discussed. PMID- 19547438 TI - Simple technique to achieve fast light in gain regime using Brillouin scattering. AB - We describe a novel technique based on stimulated Brillouin scattering for propagating fast light (signal advancement) with low distortion in optical fibers. The essence of the technique relies on the presence of two separate gain resonances in the Brillouin gain spectrum generated by cascading two different fiber segments showing distinct Brillouin shifts. It can be shown that in between these two gain spectra, a reduced group index can be obtained. To further optimize our results, we broadened the pump spectrum by introducing a modulation of the current driving the pump laser to achieve a delay-bandwidth product close to the optimum conditions. This scheme eliminates the need of an external optical modulator and offers the advantage of a much reduced signal distortion. PMID- 19547439 TI - Doppler-broadened fiber-laser-based NICE-OHMS - Improved detectability. AB - The performance of fiber-laser-based noise-immune cavity-enhanced optical heterodyne molecular spectrometry (NICE-OHMS) has been improved by elimination of the technical constraints that limited its first demonstration. Doppler-broadened detection of C(2)H(2) and CO(2) at ~1531 nm is demonstrated using a cavity with a finesse of 4800. Frequency and wavelength modulated detection at absorption and dispersion phase are compared and the optimum mode of detection is discussed. A minimum detectable absorption of 8 x 10-(11) cm(-1), which corresponds to a detection limit of 4.5 ppt (2 ppt.m) for C(2)H(2), was obtained for an acquisition time of 0.7 s by lineshape fitting. The linearity of the pressure dependence of the signal strengths is investigated for both C(2)H(2) and CO(2). PMID- 19547440 TI - Simultaneous dual-band ultra-high resolution optical coherence tomography. AB - Ultra-high resolution optical coherence tomography (OCT) imaging is demonstrated simultaneously at 840 nm and 1230 nm central wavelength using an off-the-shelf turn-key supercontinuum light source. Spectral filtering of the light source emission results in a double peak spectrum with average powers exceeding 100 mW and bandwidths exceeding 200 nm for each wavelength band. A free-space OCT setup optimized to support both wavelengths in parallel is introduced. OCT imaging of biological tissue ex vivo and in vivo is demonstrated with axial resolutions measured to be < 2 mum and < 4 mum at 840 nm and 1230 nm, respectively. This measuring scheme is used to extract spectroscopic features with outstanding spatial resolution enabling enhanced image contrast. PMID- 19547441 TI - Rapid thermal annealing in high repetition rate ultrafast laser waveguide writing in lithium niobate. AB - For the first time to our knowledge, bulk modification of lithium niobate using high repetition rate ultrashort laser pulses has been studied. A fiber based ultrafast laser has been applied in a range of 0.1 to 1.5 MHz repetition rate to directly inscribe optical waveguides in z-cut lithium niobate. Circularly polarized light with stretched 600 fs pulses produced waveguides with nearly circular mode profiles that guided in the telecom band of 1300 nm. Higher laser repetition rate of 700 kHz was found to offer smooth waveguides with low propagation loss of 0.6 dB/cm, matching the best reported value so far, with the advantage of 50 fold faster writing speed. At repetition rates of 250 kHz and higher, the tracks exhibited a cladding-like modification zone that extended outside the main laser interaction volume, yielding smoother structures, despite higher net fluence delivery, providing concrete evidence of heat accumulation and thermal annealing effects. We also present the first observation of periodic micro-structures in the bulk laser interaction volume of a non-glass material. PMID- 19547442 TI - Ultra-wide bandpass filter based on long-period fiber gratings and the evanescent field coupling between two fibers. AB - We demonstrate a fiber-based bandpass filter with an ultra-wide spectral bandwidth. The ultra-wide band feature is achieved by inscribing a long-period fiber grating (LPG) in a specially-designed low index core single mode fiber. To get the bandpass function, the evanescent field coupling between two attached fibers is utilized. By applying strain, the spectral shape of the pass-band is adjusted to flat-top and Gaussian shapes. For the flat-top case, the bandwidth is obtained ~ 160 nm with an insertion loss of ~ 2 dB. With strain, the spectral shape is switched into a Gaussian one, which has ~ 120 nm FWHM and ~ 4.18 dB insertion loss at the peak. PMID- 19547443 TI - Nonlinear diffractive optical elements. AB - We propose diffractive optical elements with a spatially-varying nonlinear refractive index. Such a component acts as a diffractive optical element whose properties depend on the intensity of the incoming beam. We present a method for designing such elements, and as specific examples we study three types of nonlinear diffractive optical elements: Nonlinear Fresnel Zone Plates, Two-foci Nonlinear Fresnel Zone Plate, and Fresnel Zone Plate to Grating interpolator. PMID- 19547444 TI - General properties of slow-plasmon resonant nanostructures: nano-antennas and resonators. AB - General properties of retardation-based resonances involving slow surface plasmon polariton (SPP) modes supported by metal nanostructures are considered. Explicit relations for the dispersion of SPP modes propagating along thin metal strips embedded in dielectric and in narrow gaps between metal surfaces are obtained. Strip and gap subwavelength resonant structures are compared with respect to the achievable scattering and local-field enhancements lending thereby their distinction as nano-antennas and nano-resonators, respectively. It is shown that, in the limit of extremely thin strips and narrow gaps, both structures exhibit the same Q factor of the resonance which is primarily determined by the complex dielectric function of metal. PMID- 19547445 TI - Simultaneous ultrafast optical pulse train bursts generation and shaping based on Fourier series developments using superimposed fiber Bragg gratings. AB - We propose an all-fiber method for the generation of ultrafast shaped pulse train bursts from a single pulse based on Fourier Series Developments (FDSs). The implementation of the FSD based filter only requires the use of a very simple non apodized Superimposed Fiber Bragg Grating (S-FBG) for the generation of the Shaped Output Pulse Train Burst (SOPTB). In this approach, the shape, the period and the temporal length of the generated SOPTB have no dependency on the input pulse rate. PMID- 19547446 TI - Confinement of band-edge modes in a photonic crystal slab. AB - We study the confinement of low group velocity band-edge modes in a photonic crystal slab. We use a rigorous, three dimensional, finite-difference time-domain method to compute the electromagnetic properties of the modes of the photonic structures. We show that by combining a defect mode approach with the high density of states associated with bandedge modes, one can design compact, fabrication-tolerant, high-Q photonic microcavities. The electromagnetic confinement properties of these cavities can foster enhanced radiation dynamics and should be well suited for ultralow-threshold microlasers and cavity quantum electrodynamics. PMID- 19547447 TI - Widely tunable guided-mode resonance nanoelectromechanical RGB pixels. AB - Widely tunable display pixels are reported. The pixel consists of a subwavelength silicon-nitride/air membrane containing complementary fixed and mobile gratings. By altering the device refractive index profile and symmetry, using MEMS actuation methods, wavelength tuning across ~100 nm per pixel in the visible spectral region is shown to be possible. Initial results illustrating the influence of structural symmetry, pixel thickness, and polarization on the spectral response are provided. These pixels exhibit ~+/-4 degrees angular acceptance aperture. Applications in compact display systems are envisioned. PMID- 19547448 TI - Effect of speckles on the depth sensitivity of laser Doppler perfusion imaging. AB - A theoretical model is presented and experimentally validated that allows the prediction of the effect of speckles on the depth sensitivity of laser Doppler perfusion imaging. It is shown that the influence of speckles on depth sensitivity is large. In particular the sensitivity to particle motion in superficial layers is strongly beam diameter dependent: decreasing the beam diameter on the tissue surface increases the sensitivity to superficial motion to a much stronger extent than sensitivity to motion at a larger depth. This can be explained through the effect of beam diameter changes on the fractional coherence areas generated by photons with different penetration depths in the tissue. PMID- 19547449 TI - Corrugated metal-coated tapered tip for scanning near-field optical microscope. AB - This paper addresses an important issue of light throughput of a metal-coated tapered tip for scanning near-field microscope (SNOM). Corrugations of the interface between the fiber core and metal coating in the form of parallel grooves of different profiles etched in the core considerably increase the energy throughput. In 2D FDTD simulations in the Cartesian coordinates we calculate near field light emitted from such tips. For a certain wavelength range total intensity of forward emission from the corrugated tip is 10 times stronger than that from a classical tapered tip. When realized in practice the idea of corrugated tip may lead up to twice better resolution of SNOM. PMID- 19547450 TI - Modulation of coaxial modal interferometers based on long period gratings in double cladding fibers. AB - This paper reports on the dynamic modulation of coaxial interferometers based on two cascaded long period gratings written in double cladding fibers. The interferometer is modulated by a piezoelectric ceramic which stretches one the gratings at tens of kHz, the output light is intensity modulated with an efficiency of 97 %. The device operates at 1530nm, has more than 50nm bandwidth, insertion loss of 0.4 dB and a temperature drift of 0.11 nm/ degrees C. PMID- 19547451 TI - Simulations and realizations of active right-handed metamaterials with negative refractive index. AB - The theory of determining the sign of the refractive index in active materials is discussed. Animations of numerical simulations are presented, supporting the claim that negative refractive index may occur in right-handed media. An example of such a medium, in the form of a lumped circuit model with active and passive resonances, is presented. PMID- 19547452 TI - First-principles study: size-dependent optical properties for semiconducting silicon carbide nanotubes. AB - Using first-principles calculations, we investigate the effect of tube size on optical properties of the zigzag, armchair, and chiral SiC nanotubes. The results indicate that the optical spectra of SiC nanotubes are dependent on the diameter and chirality, and that optical anisotropy is observed for different light polarizations. For a given chirality of SiCNTs, redshifts or blueshifts of the peaks in the dielectric function and energy loss function with increasing tube diameter are possible due to the competition between the size effect and pi orbitals overlapping, and the shifts become smaller as the tube diameter increases. The unusual optical properties of semiconducting SiC nanotubes present an opportunity for applications in electro-optical devices. PMID- 19547453 TI - Nondispersive optical phase shifter array using microelectromechanical systems based gratings. AB - We present a broadband nondispersive optical phase shifter array based on microelectromechanical systems (MEMS) technology. The device consists of an array of micro-grating elements. Each grating element utilizes an in-plane translational motion to produce a phase shift to the diffracted light beam. The phase shift is dependent only on the ratio of the grating displacement to its period, and thus independent of incident wavelength. The proposed operation principle was demonstrated with a prototype device developed using a silicon-on insulator (SOI) micromachining process. This MEMS nondispersive phase-shifter array may be useful in many multispectral applications including broadband optical phased arrays. PMID- 19547454 TI - Stokes-space derivations of generalized Schrodinger equations for wave propagation in various fibers. AB - In this report, multiple-scale analysis (averaging) is used to derive the generalized Schrodinger equations that govern light-wave propagation in strongly birefringent, randomly-birefringent and rapidly-spun fibers. The averaging procedures are described in Jones space and Stokes space. Despite the differences between the aforementioned fibers, the Stokes-space procedures associated with them are similar, and involve only quantities whose physical significances are known. Not only does the Stokes-space formalism unify the derivations of the aforementioned Schrodinger equations, it also produces equations directly in Jones-Stokes notation, which facilitates subsequent studies of polarization effects in optical systems. PMID- 19547455 TI - Efficient slow light coupling into photonic crystals. AB - We study light coupling between two photonic crystal wave-guides, one of which supports slow light. We show theoretically that a short photonic crystal waveguide between the two that need to be coupled, can lead to a vanishingly small reflectivity. The design relies on the analogy with a lambda/4 anti reflection layer in thin-film optics.We find that some of the usual relationships between the Fresnel coefficients at an interface no longer hold. PMID- 19547456 TI - Multifocal multiphoton microscopy (MMM) at a frame rate beyond 600 Hz. AB - We introduce a multiphoton microscope for high-speed three-dimensional (3D) fluorescence imaging. The system combines parallel illumination by a multifocal multiphoton microscope (MMM) with parallel detection via a segmented high sensitivity charge-couple device (CCD) camera. The instrument consists of a Ti sapphire laser illuminating a microlens array that projects 36 foci onto the focal plane. The foci are scanned using a resonance scanner and imaged with a custom-made CCD camera. The MMM increases the imaging speed by parallelizing the illumination; the CCD camera can operate at a frame rate of 1428 Hz while maintaining a low read noise of 11 electrons per pixel by dividing its chip into 16 independent segments for parallelized readout. We image fluorescent specimens at a frame rate of 640 Hz. The calcium wave of fluo3 labeled cardiac myocytes is measured by imaging the spontaneous contraction of the cells in a 0.625 second sequence movie, consisting of 400 single images. PMID- 19547457 TI - Performance analysis of 20Gb/s RZ-DPSK non-slope matched transoceanic submarine links. AB - Direct computation of the bit-error rate (BER) and laboratory experiments are used to assess the performance of a non-slope matched transoceanic submarine transmission link operating at 20Gb/s channel rate and employing return-to-zero differential-phase shift keying (RZ-DPSK) signal modulation. Using this system as an example, we compare the accuracies of the existing theoretical approaches to the BER estimation for the RZ-DPSK format. PMID- 19547458 TI - Poincare sphere representation for the anisotropy of phase singularities and its applications to optical vortex metrology for fluid mechanical analysis. AB - A new technique for fluid mechanics measurement is proposed that makes use of the elliptic anisotropy of phase singularities in the complex signal representation of a speckle-like pattern. Based on the formal analogy between the polarization field of a vector wave and the gradient field of the complex signal, the Poincare sphere representation has been used to characterize the phase singularities that serve as unique fingerprints attached to the seeding particles moving with the flow. Experimental results for flow velocity and acceleration measurement are presented that demonstrate the validity of the proposed optical vortex metrology for fluid mechanics measurement. PMID- 19547459 TI - The interaction of lipopolysaccharide with membrane receptors on macrophages pretreated with extract of Reishi polysaccharides measured by optical tweezers. AB - Lipopolysaccharide (LPS), one of the cell wall components of Gram-negative bacteria, is recognized by and interacted with receptors on macrophages. In this paper, we report the trapping of LPS-coated polystyrene particles via optical tweezers and measured its interaction with murine macrophages (J774A.1 cells) for cells pre-treated with extract of Reishi polysaccharides (EORP) vs. those without EORP treatment. Our experimental results indicate that the cellular affinity for LPS increases when the macrophage is pretreated with EORP. We demonstrate for the first time by conventional biological methods and by tracking the dynamics of optically-trapped LPS-coated particles interacting with J774A.1 cells, that EORP not only enhances J774A.1 cells surface expression of TLR4 and CD14, two receptors on macrophages, as well as LPS binding and phagocytosis internalization, but also reduces the adhesion time constant and increases the force constant of the binding interaction. The application of optical tweezers allows us to study the effect on a single cell quantitatively in real-time with a spatial resolution ~ 1 mum within a single cell. PMID- 19547460 TI - Stable operation of mode-locked fiber lasers: similariton regime. AB - A theoretical analysis of the solitary pulse parameters of the Ginzburg-Landau distributed model of a mode-locked laser is presented. For a stable operation, the mode-locked gain (g-l) < 0 is optimized relative to the chirp and a stable operation point is found in the normal dispersion regime. When the energy in the pulse is optimized relative to the mode-locking parameter, a stable point is also found in the normal dispersion regime. The two opmimizations yield the same pulse parameters when the chirp is very large which are the characteristics of the similariton regime. PMID- 19547461 TI - Theoretical and computational concepts for periodic optical waveguides. AB - We present a general, rigorous, modal formalism for modeling light propagation and light emission in three-dimensional (3D) periodic waveguides and in aggregates of them. In essence, the formalism is a generalization of well-known modal concepts for translation-invariant waveguides to situations involving stacks of periodic waveguides. By surrounding the actual stack by perfectly matched layers (PMLs) in the transverse directions, reciprocity considerations lead to the derivation of Bloch-mode orthogonality relations in the sense of E x H products, to the normalization of these modes, and to the proof of the symmetrical property of the scattering matrix linking the Bloch modes. The general formalism, which rigorously takes into account radiation losses resulting from the excitation of radiation Bloch modes, is implemented with a Fourier numerical approach. Basic examples of light scattering like reflection, transmission and emission in periodic-waveguides are accurately resolved. PMID- 19547462 TI - Single mode, polarization-independent submicron silicon waveguides based on geometrical adjustments. AB - In this work, we demonstrate via computer simulation the single mode and zero birefringence conditions for photonic wires with height and width less than 600 nm. We report on the simulation conditions for both single mode and zero birefringence in silicon-on-insulator photonic wires and sub-micron rib waveguides using a 3-dimensional imaginary beam propagation method. The results show that operation in both single mode and zero birefringence is possible under certain circumstances and that the conditions are restricted by fabrication processes where birefringence is strongly dependent upon waveguide dimensions. A matrix of waveguide parameters has been identified at both operating wavelengths of 1310 nm and 1550 nm, which can satisfy single mode and zero birefringence conditions simultaneously. This is to provide a general design rule for waveguides in small dimensions on the order of hundreds of nanometres. PMID- 19547463 TI - Measurement of the group dispersion of the fundamental mode of holey fiber by white-light spectral interferometry. AB - We present a new method for measuring the group dispersion of the fundamental mode of a holey fiber over a wide wavelength range by white-light interferometry employing a low-resolution spectrometer. The method utilizes an unbalanced Mach Zehnder interferometer with a fiber under test placed in one arm and the other arm with adjustable path length. A series of spectral signals are recorded to measure the equalization wavelength as a function of the path length, or equivalently the group dispersion. We reveal that some of the spectral signals are due to the fundamental mode supported by the fiber and some are due to light guided by the outer cladding of the fiber. Knowing the group dispersion of the cladding made of pure silica, we measure the wavelength dependence of the group effective index of the fundamental mode of the holey fiber. Furthermore, using a full-vector finite element method, we model the group dispersion and demonstrate good agreement between experiment and theory. PMID- 19547464 TI - Collective oscillations in optical matter. AB - Atom and nanoparticle arrays trapped in optical lattices are shown to be capable of sustaining collective oscillations of frequency proportional to the strength of the external light field. The spectrum of these oscillations determines the mechanical stability of the arrays. This phenomenon is studied for dimers, strings, and two-dimensional planar arrays. Laterally confined particles free to move along an optical channel are also considered as an example of collective motion in partially-confined systems. The fundamental concepts of dynamical response in optical matter introduced here constitute the basis for potential applications to quantum information technology and signal processing. Experimental realizations of these systems are proposed. PMID- 19547466 TI - Terahertz wave polarization rotation with double layered metal grating of complimentary chiral patterns. AB - We propose and demonstrate polarization rotation of a terahertz (THz) electromagnetic wave by using two-dimensional gratings consisting of two displaced layers of gold film with complimentary chiral patterns with four-fold symmetry. We develop a time domain THz polarimetry method with three wire grid polarizers and distinguish optical activity from optical anisotropy. We obtain the isotropic polarization rotation of a terahertz wave free from the birefringence of the structures. Results indicate the possibility of controlling THz polarization with artificial chiral structures fabricated with thin metal films. PMID- 19547465 TI - Image reconstruction for bioluminescence tomography from partial measurement. AB - The bioluminescence tomography is a novel molecular imaging technology for small animal studies. Known reconstruction methods require the completely measured data on the external surface, although only partially measured data is available in practice. In this work, we formulate a mathematical model for BLT from partial data and generalize our previous results on the solution uniqueness to the partial data case. Then we extend two of our reconstruction methods for BLT to this case. The first method is a variant of the well-known EM algorithm. The second one is based on the Landweber scheme. Both methods allow the incorporation of knowledge-based constraints. Two practical constraints, the source non negativity and support constraints, are introduced to regularize the BLT problem and produce stability. The initial choice of both methods and its influence on the regularization and stability are also discussed. The proposed algorithms are evaluated and validated with intensive numerical simulation and a physical phantom experiment. Quantitative results including the location and source power accuracy are reported. Various algorithmic issues are investigated, especially how to avoid the inverse crime in numerical simulations. PMID- 19547467 TI - Highly efficient four-wave parametric amplification in transparent bulk Kerr medium. AB - We report on highly efficient four-wave optical parametric amplification in a water cell pumped by an elliptically shaped, ultrashort pulsed laser beam under non-collinear phase-matching configuration. Energy conversion from pump to parametric waves as high as 25 % is obtained owing to the achievement of 1 dimensional spatial-soliton regime, which guarantees high intensity over a large interaction length and ensures high beam quality. PMID- 19547468 TI - Electromagnetic cloaking by layered structure of homogeneous isotropic materials. AB - Electromagnetic invisibility cloak requires material with anisotropic distribution of the constitutive parameters as first proposed by Pendry et al. [Science 312, 1780 (2006)]. In this paper, we proposed an electromagnetic cloak structure that does not require metamaterials with subwavelength structured inclusions to realize the anisotropy or inhomogeneity of the material parameters. We constructed a concentric layered structure of alternating homogeneous isotropic materials that can be treated as an effective medium with the required radius-dependent anisotropy. With proper design of the permittivity or the thickness ratio of the alternating layers, we demonstrated the low-reflection and power-flow bending properties of the proposed cloaking structure through rigorous analysis of the scattered electromagnetic fields. The proposed cloaking structure could be possibly realized by normal materials, therefore may lead to a practical path to an experimental demonstration of electromagnetic cloaking, especially in the optical range. PMID- 19547469 TI - Performance comparison of 40 Gb/s ULH transmissions using CSRZ-ASK or CSRZ-DPSK modulation formats on UltraWave fiber (TM) fiber. AB - In this work we present extensive comparisons between numerical modelling and experimental measurements of the transmission performance of either CSRZ-ASK or CSRZ-DPSK modulation formats for 40-Gb/s WDM ULH systems on UltraWave(TM) fiber spans with all-Raman amplification. We numerically optimised the amplification and the signal format parameters for both CSRZ-DPSK and CSRZ-ASK formats. Numerical and experimental results show that, in a properly optimized transmission link, the DPSK format permits to double the transmission distance (for a given BER level) with respect to the ASK format, while keeping a substantial OSNR margin (on ASK modulation) after the propagation in the fiber line. Our comparison between numerical and experimental results permits to identify what is the most suitable BER estimator in assessing the transmission performance when using the DPSK format. PMID- 19547470 TI - Magnetically actuated MEMS microlens scanner for in vivo medical imaging. AB - A magnetically actuated MEMS scanner with a microfabricated ferromagnetic nickel platform and thermosetting polydimethylsiloxane (PDMS) microlens is demonstrated. The device is driven by an external AC magnetic field, eliminating chip circuitry and thermal deformation from joule heating. The resonant frequency of 215.2 Hz and scanning angle of 23 of the scanner have been demonstrated. Experimental studies and optical modeling have shown that this microlens scanner achieves a scanning range of 125 mum when actuated by an external magnetic field of 22.2x10 3 Tesla flux density. The device has potential applications in in vivo medical imaging for minimally invasive diagnoses. PMID- 19547471 TI - In vivo and ex vivo imaging of intra-tissue elastic fibers using third-harmonic generation microscopy. AB - Elastin is an essential and widespread structural protein in charge of the integrity on tissues and organs. In this study, we demonstrate that elastin is a major origin of the third-harmonic-generation (THG) contrast under Cr:forsterite laser excitation operating at 1230nm, with selective visualization inside many tissues such as lung tissues and arteries. In vivo imaging of the nude mouse elastic cartilage beneath the hypodermis by epi- THG microscopy keeps the high resolution and contrast in all three dimensions. Combined with second-harmonic generation microscopy, THG microscopy exhibits the ability to show the extraordinary proliferation of elastic fibers for the ophthalmic disease of pterygium and the capability of distinguishable visualization from collagen. PMID- 19547472 TI - Monitoring thermally-induced phase transitions in porcine cornea with the use of fluorescence micro-imaging analysis. AB - Thermal modifications induced in corneal stroma were investigated with the use of fluorescence microscopy. Tissue samples were heated in a water bath at temperatures in the 35-90 degrees C range. Fluorescence images of the structural modifications induced were acquired after staining with Indocyanine Green (ICG). Discrete Fourier Transform (DFT) and entropy analyses of each image made it possible to characterize the thermally-induced phase transitions in the stroma, and to indicate a threshold value for high thermal damage. The procedure could be proposed as the basis for a real-time controlling system for surgical techniques based on induced thermal effects. PMID- 19547473 TI - Induced modulation instability and recurrence in nematic liquid crystals. AB - We study induced modulation instability in a nematic liquid crystal cell. Two broad elliptical beams along one direction are launched into the cell. The two beams have slightly different angle in order to create a sinusoidally varying intensity at the entrance of the cell. In this way, the gain of perturbations with different spatial frequency is investigated. The evolution of the optical pattern, for certain conditions, shows a recurrence of the signal. We believe that this is the manifestation of the Fermi-Pasta- Ulam recurrence and to the best of our knowledge, the first experimental observation of this phenomenon in the spatial optical domain. Numerical simulations show a good agreement with the experimental findings. PMID- 19547474 TI - Optical sectioning microscopes with no moving parts using a micro-stripe array light emitting diode. AB - We describe an optical sectioning microscopy system with no moving parts based on a micro-structured stripe-array light emitting diode (LED). By projecting arbitrary line or grid patterns onto the object, we are able to implement a variety of optical sectioning microscopy techniques such as grid-projection structured illumination and line scanning confocal microscopy, switching from one imaging technique to another without modifying the microscope setup. The micro structured LED and driver are detailed and depth discrimination capabilities are measured and calculated. PMID- 19547475 TI - Liquid-core, liquid-cladding photonic crystal fibers. AB - We experimentally demonstrate a simple and novel technique to simultaneously insert a liquid into the core of a hollow-core photonic crystal fiber (PCF) and a different liquid into its cladding. The result is a liquid-core, liquid-cladding waveguide in which the two liquids can be selected to yield specific guidance characteristics. As an example, we tuned the core-cladding index difference by proper choice of the inserted liquids to obtain control over the number of guided modes. Single-mode guidance was achieved for a particular choice of liquids. We also experimentally and theoretically investigated the nature of light confinement and observed the transition from photonic bandgap to total internal reflection guidance both with the core-cladding index contrast and with the PCF length. PMID- 19547476 TI - Linearly polarized emission from GaInN lightemitting diodes with polarization enhancing reflector. AB - A polarization-enhancing reflector design, which is matched to the emission characteristics of GaInN/GaN 460 nm light-emitting diodes grown on (0001) oriented sapphire substrates, is reported. Side-emitted light from these devices is known to be highly polarized with the electric field in the plane of the active region. Through selective rotation of polarization by the reflector, the in-plane polarized side-emitted light is directed upwards with a single dominant linear polarization. Polarization ratios as high as 3.5:1 are measured in the farfield, and the average polarization ratio is 1.9:1. If only light that strikes the reflector is considered, the polarization ratio is 2.5:1. The concept of the polarization-enhancing reflector and the numerical algorithm used to generate the optimized shape are also described. PMID- 19547477 TI - Mid-infrared methane detection in a photonic bandgap fiber using a broadband optical parametric oscillator. AB - We demonstrate methane sensing using a photonic bandgap fiber-based gas cell and broadband idler pulses from a periodically-poled lithium niobate femtosecond optical parametric oscillator. The hollow core of the fiber was filled with a methane:nitrogen mixture, and Fourier transform spectroscopy was used to measure transmission spectra in the 3.15-3.35 mum methane absorption region. The method has applications in gas sensing for remote or hazardous environments and potentially at very low concentrations. PMID- 19547478 TI - Mode-locked silicon evanescent lasers. AB - We demonstrate electrically pumped lasers on silicon that produce pulses at repetition rates up to 40 GHz. The mode locked lasers generate 4 ps pulses with low jitter and extinction ratios above 18 dB, making them suitable for data and telecommunication transmitters and for clock generation and distribution. Results of both passive and hybrid mode locking are discussed. This type of device could enable new silicon based integrated technologies, such as optical time division multiplexing (OTDM), wavelength division multiplexing (WDM), and optical code division multiple access (OCDMA). PMID- 19547479 TI - Polarization-Conversion Guided Mode (PCGM) technique for exploring thin anisotropic surface layers. AB - A Polarization-Conversion Guided Mode (PCGM) technique has been developed to quantify optical anisotropy as low as 10-5 for a surface layer only 10 nm thick. The optical geometry consists of an index-fluid matched prism-coupler and an air gap waveguide comprising the thin sample on a glass plate as the incident surface with a gold reflector forming the other surface of the guide. This allows non destructive characterization of the optical anisotropy of surface layers. The polarization conversion signal is extraordinarily sensitive. Thus the influence of the polarization purity of the incoming beam, very small twists and/or tilts between the normal to the prism bottom surface and the sample plane, have all been analyzed in detail to allow extraction of the sought for information about the thin layer. Rubbed polyimide thin films and incline-evaporated SiOx layers, both used for liquid crystal alignment, have been examined by this PCGM technique to demonstrate its power. PMID- 19547480 TI - LD light antisqueezing through fiber propagation in reflection-type interferometer. AB - Antisqueezed light is a possible resource to apply quantum information technologies to the real world. When antisqueezed light is used in secure optical communications, an LD is a preferable light source from an engineering point of view. Although LD output power is low, LD light can be antisqueezed with the help of an EDFA in a reflection-type interferometer consisting of a standard single mode fiber of typically 5 km. The ellipticity of the obtained antisqueezed light was 9 at maximum in a balanced interferometer case, and the angle that was subtended by antisqueezed fluctuations at the origin of phase space was 23 degrees at maximum. The feasibility of secure optical communications using antisqueezed light is demonstrated. PMID- 19547481 TI - Enhanced thermal emission from individual antenna-like nanoheaters. AB - Here we report polarization-sensitive, thermal radiation measurements of individual, antenna-like, thin film Platinum nanoheaters. These heaters confine the lateral extent of the heated area to dimensions smaller (or comparable) to the thermal emission wavelengths. For very narrow heater structures the polarization of the thermal radiation shows a very high extinction ratio as well as a dipolar-like angular radiation pattern. A simple analysis of the radiation intensities suggests a significant enhancement of the thermal radiation for these very narrow heater structures. PMID- 19547482 TI - Cryogenic temperature characteristics of Verdet constant on terbium gallium garnet ceramics. AB - As the first demonstration of Faraday effect in a TGG ceramics, its Verdet constant at 1053 nm is evaluated to be 36.4 rad/Tm at room temperature which is same as that of the single crystal. In addition, the temperature dependence of Verdet constant is obtained experimentally. At liquid helium temperature, it is 87 times greater than that at room temperature. PMID- 19547483 TI - Intra-cavity absorption spectroscopy with narrow-ridge microfluidic quantum cascade lasers. AB - We demonstrate microfluidic laser intra-cavity absorption spectroscopy with mid infrared lambda approximately 9mum quantum cascade lasers. A deepetched narrow ridge waveguide laser is placed in a microfluidic chamber. The evanescent tails of the laser mode penetrate into a liquid on both sides of the ridge. The absorption lines of the liquid modify the laser waveguide loss, resulting in significant changes in the laser emission spectrum and the threshold current. A volume of liquid as small as ~10pL may, in principle, be sufficient for sensing using the proposed technique. This method, similar to the related gas-phase technique, shows promise as a sensitive means of detecting chemicals in small volumes of solutions. PMID- 19547484 TI - Tensile-strained, n-type Ge as a gain medium for monolithic laser integration on Si. AB - We analyze the optical gain of tensile-strained, n-type Ge material for Si compatible laser applications. The band structure of unstrained Ge exhibits indirect conduction band valleys (L) lower than the direct valley (Gamma) by 136 meV. Adequate strain and n-type doping engineering can effectively provide population inversion in the direct bandgap of Ge. The tensile strain decreases the difference between the L valleys and the Gamma valley, while the extrinsic electrons from n-type doping fill the L valleys to the level of the Gamma valley to compensate for the remaining energy difference. Our modeling shows that with a combination of 0.25% tensile strain and an extrinsic electron density of 7.6x10(19)/cm(3) by n-type doping, a net material gain of ~400 cm(-1) can be obtained from the direct gap transition of Ge despite of the free carrier absorption loss. The threshold current density for lasing is estimated to be ~6kA cm(-2) for a typical edgeemitting double heterojunction structure. These results indicate that tensile strained n-type Ge is a good candidate for Si integrated lasers. PMID- 19547485 TI - Bandpass filters based on phase-shifted photonic crystal waveguide gratings. AB - In this paper, a bandpass transmission filter realized in phase-shifted waveguide gratings based on photonic crystals (PCs) is proposed. Phase-shift regions each composed of one period of photonic crystal (PC) waveguide are incorporated into PC waveguide gratings. The magnitudes of the phase-shifts are modified by involving small changes in the size of the border rods in the phase-shift regions. Using standard coupled-mode theory and finite-difference time-domain (FDTD) method, we show that by properly choosing the magnitudes of phase-shifts and the lengths of waveguide gratings, a flat-top and sharp roll-off response with a narrow bandwidth is theoretically and numerically achieved by the designed filter. A further analysis shows that the center frequency of the transmission band can be changed by altering the magnitude of the phase-shift and the response performance exhibits relaxed sensitivity to the phase-shift variation. As a specific application, we theoretically demonstrate a third-order Chebyshev bandpass filter based on compound phase-shifted PC waveguide gratings. The filter performance is suitable for dense wavelength-division-multiplexed (DWDM) optical communication systems with a channel spacing of 100-GHz. PMID- 19547486 TI - Noise pre-filtering techniques in fluorescence-enhanced optical tomography. AB - In this contribution, different measurement noise pre-filtering techniques were developed using frequency-domain fluorescence measurements of homogeneous breast phantoms. We demonstrated that implementing noise pre-filtering, based on modulation depth and measurement error in amplitude, can improve model match between experimental and simulated data under varying experimental conditions (target depths, 1-3 cm and fluorescence optical contrast, 1:0 and 100:1). Noise pre-filtering also improves the qualitative estimation of target(s) location in reconstructed images in deep target(s) when there was fluorescence in the background. Interestingly, decreases in model mismatch did not necessarily correlate with increases in reconstructed target accuracy. In addition, it was observed that pre-filtering measurement noise using different criteria can help differentiate target(s) from artifacts, thus possibly minimizing the false positive cases in a clinical environment. PMID- 19547488 TI - Hyperspectral imaging of plasmonic nanostructures with nanoscale resolution. AB - We report on the first realization of a hyperspectral imaging technique for surface plasmon polaritons on metallic nanostructures. The technique uses a scanning electron beam and allows for simple visualization of light emission from decoupled plasmons, providing information on decay lengths and feature sizes with nanometer resolution. PMID- 19547487 TI - Spatial-temporal structure of the femtosecond third harmonic generation in photonic-crystal fibers. AB - We analyze the third harmonic generation by the supercontinuum at 1.5 mum in nonlinear-glass microstructured fibers. The numerical model includes the real field dynamics and allows analyzing the spectral as well as the temporal structure of the generated field. The calculated third harmonic and supercontinuum spectra are compared with the experiment in the SF6 glass PCF pumped by a femtosecond Cr(4+):YAG oscillator. The spectral structure of the third harmonic is composed of the 10-20 high-order modes excited at the phase matching points around 500-550 nm. The individual third harmonic modes have spectral widths of 2-4 nm. In the time domain, the third harmonic signal is a ~10 ps pulse with ps-scale slow modulation, containing subpicosecond ripples at its trailing edge. PMID- 19547489 TI - Young's interference of double metallic nanoslit with different widths. AB - A type of metallic double nanoslit with different widths is proposed to investigate Young's interference mediated by surface plasmon polaritons (SPPs). Numerical calculations show that the Young's interference order could be shifted readily by adjusting the width difference between two slits. Further calculations indicate that the interference order shift related to the additional phase retardation is caused by the distinct surface plasmon mode in two slits. Since the surface plasmon mode in a nanoslit is extremely sensitive to the incident wavelength, it suggests a potential way of ultrahigh resolution spectral analysis via measuring the shift of Young's interference. PMID- 19547490 TI - Liquid crystal lens with large focal length tunability and low operating voltage. AB - We demonstrate a tunable focus liquid crystal (LC) lens by sandwiching a homogeneous LC layer between a planar electrode and a curved electrode. The curved electrode which is made of conductive polymer has parabolic shape with a large apex distance. Such design dramatically reduces the phase loss which leads to a short focal length (~15 cm). By using a thin top glass substrate on the curved electrode side, the operating voltage of the lens cell is reduced to ~23 V(rms). This LC lens has advantages in wide focal length tunability, low operating voltage, and good mechanical stability. PMID- 19547491 TI - Efficient narrow-linewidth volume-Bragg grating-locked Nd:fiber laser. AB - A neodymium doped microstructured large mode area fiber is frequency locked with a volume Bragg grating. This configuration is compared with a conventional fiber laser setup. A high efficiency (51% slope), stable output and a drastically narrowed linewidth (<0.07nm) are achieved. PMID- 19547492 TI - Single-shot, space- and time-resolved measurement of rotational wavepacket revivals in H(2), D(2), N(2), O(2), and N(2)O. AB - Femtosecond laser-induced alignment and periodic recurrences in hydrogen and deuterium are measured in a single shot for the first time, in a room temperature gas cell. Single-shot Supercontinuum Spectral Interferometry (SSSI) is employed, with measurements also performed in room temperature samples of nitrogen, oxygen, and nitrous oxide. Unlike previous optical techniques for probing molecular alignment in gases or liquids, SSSI quantitatively and directly measures the degree of molecular alignment without reliance on model fits, and it can do so with spatial resolution transverse to the pump beam. In addition, wavepacket collisional dephasing rates can be directly measured in gas samples at useful densities. PMID- 19547493 TI - High spectral power density supercontinuum generation in a nonlinear fiber amplifier. AB - We present an experimental study on supercontinuum generation with high spectral power density by using a commercial nonlinear fiber amplifier. This new approach consists in the simultaneous combination of the amplification of a pulsed seed signal at 1.06 microm and its peak-power-induced spectral broadening as the optical pulse propagates along the amplifying fiber. A 750-nm broadening from 1 microm to 1.75 microm with tunable spectral power density according to the amplifier gain level is obtained. Spectral power density in excess of 3 mW/nm is demonstrated. PMID- 19547494 TI - High-power 894 nm monolithic distributed-feedback laser. AB - A ridge-waveguide InGaAs/GaAsP laser, emitting up to 250 mW in a single lateral and longitudinal mode at a wavelength of 894 nm, is presented. The distributed feedback is provided by a second order grating, formed into an InGaP/GaAs/InGaP multilayer structure. Owing to the stable lasing frequency, the large side mode suppression ratio (> 40 dB) and small spectral line width (< 200 kHz) the diode laser is well suited for caesium D1 spectroscopy. This was verified by the measurement of the hyperfine structure of the D1 line. PMID- 19547495 TI - Wide frequency range measurements of absolute phase and amplitude of vibrations in micro- and nanostructures by optical interferometry. AB - A heterodyne interferometer has been built in order to characterize vibrations on micro- and nanostructures. The interferometer offers the possibility of both absolute phase and high resolution absolute amplitude vibrational measurements. By using two acousto-optic modulators (AOMs) in one of the interferometer arms and varying the frequency inputs of both, the setup is designed to measure vibrations in the entire frequency range 0 - 1.2GHz. The system is here demonstrated on Capacitor Micromachined Ultrasonic Transducers (CMUTs) and a PZT transducer to show measurements from 5kHz up to 35MHz. We have measured absolute amplitudes with picometer resolution. PMID- 19547496 TI - High bandwidth absorption spectroscopy with a dispersed supercontinuum source. AB - An optical gas sensor is presented, making use of a dispersed supercontinuum source, capable of acquiring broad bandwidth spectra at ultrahigh wavelength sweep and repetition rates. Wavelength sweeps from 1100 nm to 1700 nm can be performed in 800 ns at a spectral resolution of 40 pm. This is comparable to line widths of molecular spectra at atmospheric pressure. Quantitative measurements are presented of CH(4) employing 80 nm wide sweeps over the P- Q- and R-branches of the 2nu(3) transition near 1665 nm, at rates exceeding 100 kHz. The effective acquisition rate is determined by the amount of averaging required, and the effect of this averaging on observed precision is investigated. PMID- 19547497 TI - Optical frequency comb generator based on actively mode-locked fiber ring laser using an acousto-optic modulator with injection-seeding. AB - We present an optical frequency comb generator (OFCG) based on a fiber ring laser that provides a bandwidth of over 1.8 THz with mode-locked pulse operation by external injection seeding. The OFCG was developed via a configuration with actively mode-locked fiber ring laser utilizing an acousto-optic modulator (AOM) with a carrier frequency of 150 MHz when an external seeding laser was injected into the fiber ring cavity. To our knowledge, our experimental device has the widest comb bandwidth ever reported for an OFCG based on a fiber ring loop adopting an AOM device, and ours is the first device that can operate with an actively mode-locked scheme. PMID- 19547498 TI - Mapping of cerebro-vascular blood perfusion in mice with skin and skull intact by Optical Micro-AngioGraphy at 1.3 mum wavelength. AB - Optical micro-angiography (OMAG) was developed to achieve volumetric imaging of the microstructures and dynamic cerebrovascular blood perfusion in mice with capillary level resolution and high signal-to-background ratio. In this paper, we present a high-speed and high-sensitivity OMAG imaging system by using an InGaAs line scan camera and broadband light source at 1.3 mum wavelength for enhanced imaging depth in tissue. We show that high quality imaging of cerebrovascular blood perfusion down to capillary level resolution with the intact skin and cranium are obtained in vivo with OMAG, without the interference from the blood perfusion in the overlaying skin. The results demonstrate the potential of 1.3 mum OMAG for high-speed and high-sensitivity imaging of blood perfusion in human and small animal studies. PMID- 19547499 TI - Photonic bandgap fiber-based Surface Plasmon Resonance sensors. AB - The concept of photonic bandgap fiber-based surface plasmon resonance sensor operating with low refractive index analytes is developed. Plasmon wave on the surface of a thin metal film embedded into a fiber microstructure is excited by a leaky Gaussian-like core mode of a fiber. We demonstrate that by judicious design of the photonic crystal reflector, the effective refractive index of the core mode can be made considerably smaller than that of the core material, thus enabling efficient phase matching with a plasmon, high sensitivity, and high coupling efficiency from an external Gaussian source, at any wavelength of choice from the visible to near-IR. To our knowledge, this is not achievable by any other traditional sensor design. Moreover, unlike the case of total internal reflection waveguide-based sensors, there is no limitation on the upper value of the waveguide core refractive index, therefore, any optical materials can be used in fabrication of photonic bandgap fiber-based sensors. Based on numerical simulations, we finally present designs using various types of photonic bandgap fibers, including solid and hollow core Bragg fibers, as well as honeycomb photonic crystal fibers. Amplitude and spectrum based methodologies for the detection of changes in the analyte refractive index are discussed. Furthermore, sensitivity enhancement of a degenerate double plasmon peak excitation is demonstrated for the case of a honeycomb fiber. Sensor resolutions in the range 7 * 10(-6) -5 * 10(-5) RIU were demonstrated for an aqueous analyte. PMID- 19547500 TI - Unidirectional excitation of surface plasmons by slanted gratings. AB - Surface plasmon excitation by normally incident light on surface-relief metallic diffraction gratings is studied numerically. Predominantly unidirectional excitation is achieved with a grating of either a slanted lamellar or an inclined sinusoidal groove profile, both having shallow depths. Maps of Poynting vector illustrate that the energy flow turns from normal incidence in the far-field region to a pattern almost parallel to the grating surface in the required direction of excitation of a single SPP wave. PMID- 19547501 TI - Numerical investigation of field enhancement by metal nano-particles using a hybrid FDTD-PSTD algorithm. AB - We present a novel numerical scheme for the simulation of the field enhancement by metal nano-particles in the time domain. The algorithm is based on a combination of the finite-difference time-domain method and the pseudo-spectral time-domain method for dispersive materials. The hybrid solver leads to an efficient subgridding algorithm that does not suffer from spurious field spikes as do FDTD schemes. Simulation of the field enhancement by gold particles shows the expected exponential field profile. The enhancement factors are computed for single particles and particle arrays. Due to the geometry conforming mesh the algorithm is stable for long integration times and thus suitable for the simulation of resonance phenomena in coupled nano-particle structures. PMID- 19547502 TI - Understanding laser stabilization using spectral hole burning. AB - There have recently been several studies of the performance of laser frequency stabilization using spectral holes in solids, instead of an external cavity, as a frequency reference. Here an analytical theory for Pound-Drever-Hall laser frequency stabilization using spectral hole-burning is developed. The interaction between the atomic medium and the phase modulated light is described using a linearized model of the Maxwell-Bloch equations. The interplay between the carrier and modulation sidebands reveals significant differences from the case of locking to a cavity. These include a different optimum modulation index, an optimum sample absorption, and the possibility to lock the laser in an inherent linear frequency drift mode. Spectral holes in solids can be permanent or transient. For the materials normally used, the dynamics and time scales of transient holes often depend on population relaxation processes between ground state hyperfine levels. These relaxation rates can be very different for different solid state materials. We demonstrate, using radio-frequency pumping, that the hyperfine population dynamics may be controlled and tailored to give optimum frequency stabilization performance. In this way also materials with initially non-optimum performance can be used for stabilization. The theoretical predictions regarding the inherent linear frequency drift is compared to experimental data from a dye laser stabilized to a spectral hole in a Pr(3+)3+:Y(2)SiO(5) crystal. PMID- 19547503 TI - 1310nm silicon evanescent laser. AB - We report the first 1310 nm hybrid laser on a silicon substrate. This laser operates continuous wave (C.W.) up to 105 degrees C. The room temperature threshold current of this laser is 30 mA, and the maximum single sided fiber coupled output power is 5.5 mW. PMID- 19547504 TI - Emission spectrum of electromagnetic energy stored in a dynamically perturbed optical microcavity. AB - An ultrafast pump-probe experiment is performed on wavelength-scale, silicon based, optical microcavities that confine light in three dimensions with resonant wavelengths near 1.5 mum, and lifetimes on the order of 20 ps. A below-bandgap probe pulse tuned to overlap the cavity resonant frequency is used to inject electromagnetic energy into the cavity, and an above-bandgap pump pulse is used to generate free carriers in the silicon, thus altering the real and imaginary components of the cavity's refractive index, and hence its resonant frequency and lifetime. When the pump pulse injects a carrier density of ~ 5 x10(17) cm(-3) before the resonant probe pulse strikes the sample, the emitted radiation from the cavity is blue-shifted by 16 times the bare cavity linewidth, and the new linewidth is 3.5 times wider than the original. When the pump pulse injects carriers, and thus suddenly perturbs the cavity properties after the probe pulse has injected energy into the cavity, we show that the emitted radiation is not simply a superposition of Lorentzians centred at the initial and perturbed cavity frequencies. Under these conditions, a simple model and the experimental results show that the power spectrum of radiation emitted by the stored electromagnetic energy when the cavity frequency is perturbed during ring-down consists of a series of coherent oscillations between the original and perturbed cavity frequencies, accompanied by a gradual decrease and broadening of the original cavity line, and the emergence of the new cavity resonance. The modified cavity lifetime is shown to have a significant impact on the evolution of the emission as a function of the pump-probe delay. PMID- 19547505 TI - Optical near-field analysis of spherical metals: Application of the FDTD method combined with the ADE method. AB - The time-average energy density of the optical near-field generated around a metallic sphere is computed using the finite-difference time-domain method. To check the accuracy, the numerical results are compared with the rigorous solutions by Mie theory. The Lorentz-Drude model, which is coupled with Maxwell's equation via motion equations of an electron, is applied to simulate the dispersion relation of metallic materials. The distributions of the optical near filed generated around a metallic hemisphere and a metallic spheroid are also computed, and strong optical near-fields are obtained at the rim of them. PMID- 19547507 TI - Experimental observation of optical orientation of dipolar centers. AB - We report discovery of recently predicted phenomenon, optical orientation by polar way of local centers with permanent dipole moment. In a crystal of Bi(12)SiO(20) grown in the argon atmosphere electrical current arises when the polarization of incident light is periodically modulated. Dependence of the current amplitude on the modulation frequency allows us to attribute this current to the predicted effect, which is supported by the data on light-induced dichroism and photoconductivity of the sample. A model of donor-acceptor pairs as dipolar centers is shown to be able to explain main peculiarities of optical orientation of dipolar centers in the crystal. PMID- 19547506 TI - WDM signal regeneration using a single alloptical device. AB - Using the principle of quasi-continuous filtering in a non-linear fibre, we propose an optical device for the simultaneous regeneration of sevaral channels at 40 Gbit/s. Simulations predict an improvement of the signal quality for four channels by more than 6.8 dB. PMID- 19547508 TI - Retrieval of particle size distribution in the dependent model using the moment method. AB - The problem of determining particle size distribution using the moment method in the spectral extinction technique is studied. The feasibility and reliability of the retrieval of spherical particle size distribution using the moment method are investigated. The single spherical particle extinction efficiency, which is derived theoretically using the Mie's solution to Maxwell's equation, is approximated with a higher order polynomial in order to apply the moment method. Simulation and experimental results indicate that a fairly reasonable representation of the particle size distribution can be obtained using the moment method in the dependent model algorithm. The method has advantages of simplicity, rapidity, and suitability for in-line particle size measurement. PMID- 19547509 TI - Effect of the incident angle on the electric near field of a conical probe for plane wave and Gaussian beam illumination. AB - We consider a simple analytical model for the electric near field of a semi infinite conical probe and apply it to study the incident angle dependence of the field for the case of side illumination by both the plane wave and the Gaussian beam. The electric near field is shown to peak when approaching the grazing incidence. In some cases, a peak can also occur at an incident angle somewhat below 90 degrees . The results obtained are in qualitative agreement with those for a thin semi-infinite wire and previously published results for the finite size conical probes. PMID- 19547510 TI - Athermal emission in Yb,Er:glass. AB - We report on the temperature dependent spectroscopic properties of Yb(3+) and Er(3+) co-doped glass gain media in the eye-safe spectral region. Measurements suggest that judicious selection of the operating wavelength can lead to a laser output with minimal dependence on the temperature of the gain medium. PMID- 19547511 TI - Design-related losses of double-fishnet negative-index photonic metamaterials. AB - The literature regarding the influence of the hole shape on the performance, especially on the losses, of negative-index metamaterials on the basis of the so called double-fishnet structure is unclear. We investigate this aspect in a systematic theoretical study showing that the figure of merit can differ by as much as a factor of 2.5 between rectangular and circular holes, respectively. PMID- 19547512 TI - Subwavelength optical imaging of evanescent fields using reflections from plasmonic slabs. AB - Reflection can significantly improve the quality of subwavelength near-field images, which is explained by appropriate interference between forward and reflected waves. Plasmonic slabs may form approximate super-mirrors. This paper develops general theory in both spectral and spatial representations that allows the reflector position and permittivity to be determined for optimum image uniformity. This elucidates previous observations and predicts behaviour for some other interesting regimes, including interferometric lithography. PMID- 19547513 TI - Tailoring CW supercontinuum generation in microstructured fibers with two-zero dispersion wavelengths. AB - We theoretically study broadband supercontinuum generation in photonic crystal fibers exhibiting two zero dispersion wavelengths and under continuous-wave pumping. We show that when the pump wavelength is located in between the zero dispersion wavelengths, a wide and uniform spectral broadening is achieved through modulation instability, generation of both blue-shifted and red-shifted dispersive waves and subsequently through soliton self-frequency shift. This supercontinuum is therefore bounded by these two dispersive waves which allow the control of its bandwidth by a suitable tuning of the fiber dispersion. As a relevant example, we predict that broadband (1050-1600 nm) continuous-wave light can be generated in short lengths of microstructured fibers pumped by use of a 10 W Ytterbium fiber laser. PMID- 19547514 TI - Microring-based modulation and demodulation of DPSK signal. AB - Ultra-small modulator and demodulator for 10 Gb/s differential phase-shift-keying (DPSK), using silicon-based microrings, are proposed. A single-waveguide microring modulator with over-coupling between ring and waveguide generates a DPSK signal, while a double-waveguide microring filter enables balanced DPSK detection. These modulator and demodulator are characterized. A trade-off between pattern dependence of the Duobinary signal and alternate-mark inversion signal power in demodulator design is discussed. Power penalty of the proposed approach is 0.8 dB relative to baseline using conventional modulation and demodulation techniques. PMID- 19547515 TI - Enhancement of multiwavelength generation in the L-band by using a novel Brillouin-Erbium fiber laser with a passive EDF booster section. AB - We demonstrate a novel scheme to generate multiple wavelengths in the L-band using a Brillouin-Erbium fiber laser. Our scheme utilizes extraneous amplified spontaneous emission in the C-band as a secondary pump source for a passive Erbium-doped fiber gain section along the cavity. The Brillouin gain medium is generated by the 6.7 km long of single mode fiber (SMF-28). We experimentally demonstrate that a total of 28 stable output channels with a spacing of 0.089 nm can be generated using our new scheme. This represents a 33% increase in the number of wavelengths generated compared with conventional schemes. PMID- 19547517 TI - Monolithic Bragg-locked Nd:GdVO4 laser. AB - We present a monolithic single-longitudinal-mode laser based on Nd:GdVO(4) and a volume Bragg grating. The laser at 1066 nm had a bandwidth below 40 MHz at a power of 30 mW. With temperature, the laser frequency could be continuously tuned without mode hops over a range of 80 GHz. The demonstrated laser design is very compact and simple and can be used to lock the laser wavelength anywhere in the gain spectrum. PMID- 19547516 TI - Mechanisms of generation of multi-peak and mode-locked resembling pulses in Q switched Yb-doped fiber lasers. AB - The output pulse characteristics of Q-switched Yb-doped fiber lasers have been investigated experimentally. It has been observed that for any typical modulation frequency, the pump power and the modulator OFF-time govern the shape of the output Q-switched pulse. At a fixed modulation frequency, with a fine adjustment of acousto-optic modulation window ON-time, pump power and cavity mirror position, it was possible to obtain modulation free single-peak pulse, multi-peak pulse, mode-locked resembling pulse and multi-pulse structured pulse shapes in a Q-switched fiber laser output. These observations have been analyzed and explained. Our investigations show that multi-peak pulse output is due to onset of nonlinear phenomena like SBS and SRS. Similarly, we have found that the mode locked resembling periodically modulated output pulse shape is due to mode beating between the zeroeth order and the first order diffracted beams of the intra-cavity acousto-optic Q-switch. PMID- 19547518 TI - Geometric optics method for surface plasmon integrated circuits. AB - A fully-analytic expression is derived for the phase of reflection of surface plasmons at a semi-infinite boundary. We apply this phase of reflection result in the geometric optics method to calculate the properties of surface plasmon stripe waveguides. Comprehensive vectorial numerical computations are compared with the analytic method to show that it is accurate. The loss of the surface plasmon stripe waveguide is calculated using the geometric optics approach, which is also found to be in close agreement with numerical computations. The geometric optics approach may be used to obtain accurate results for other surface plasmon structures, such as microdisk or ring resonators, with greater efficiency and physical intuition than numerical computation. PMID- 19547519 TI - Wave-front reconstruction without twin-image blurring by two arbitrary step digital holograms. AB - We discuss a novel approach for numerical wave-front reconstruction which utilizes arbitrary phase step digital holography. Our experimental results demonstrate that only two digital holograms and a simple estimation procedure are required for twin-image suppression, and for numerical reconstruction. One advantage of this approach is its simplicity. Only one estimate equation needs be applied. In addition the optical system can be constructed from inexpensive, generally available elements. Another advantage is the effectiveness of the method. The tolerance of the estimated value is less than 1% different than the actual value. This means that the quality of the reconstructed image is superior. This novel approach should make the application of digital holography easier and more widely available. PMID- 19547520 TI - Enhancement and tunability of active plasmonic by multilayer grating coupled emission. AB - The effect of coupled mode surface plasmon polaritons (SPPs) on the active emission of a nanostructure grating with organic semiconductor material, Alq(3), on the surface was investigated in this study. We report surface plasmon grating coupled emission (SPGCE) from excited organic layer on metal grating in both organic/metal (2-Layer) and organic/metal/organic/metal (4-Layer) structures. The dispersion relation was obtained from angle-resolved photoluminescence measurement. The resultant emission intensity can have up to 6 times enhancement on the 4- Layer device and the Full-Width Half-Maximum (FWHM) is less than 50 nm. The combination of SPPs on organic/metal interface allows specific directional emission and color appearance of Alq(3) fluorophores. Potential applications of such an active plasmonics with enhanced resonant energy emission due to interactions on the organic/metal nano-grating as biosensor were presented and discussed. PMID- 19547521 TI - Haptic guidance for improved task performance in steering microparticles with optical tweezers. AB - We report the manipulation of 4-5 mum diameter polymer microspheres floating in water using optical tweezers (OT) and a haptic device (i.e. force-reflecting robotic arm). Trapped microspheres are steered using the end-effector of a haptic device that is virtually coupled to an XYZ piezo-scanner controlling the movements of the fluid bed. To help with the manipulations, we first calculate a collision-free path for the particle and then display artificial guidance forces to the user through the haptic device to keep him/her on this path during steering. Experiments conducted with 8 subjects show almost two-fold improvements in the average path error and average speed under the guidance of haptic feedback. PMID- 19547522 TI - Variation of Bragg condition in low-glass-transition photorefractive polymers when recorded in reflection geometry. AB - Two low-glass transition photorefractive polymer composites were investigated in a symmetric reflection geometry. The holograms recorded in 105 mum thick devices have reached diffraction efficiencies as high as 60%. Unlike the gratings recorded in transmission geometry, holograms recorded in reflection geometry showed high angular selectivity and the Bragg condition was observed to be sensitive to the magnitude of the external bias field. We attribute this effect to poling-induced birefringence and give a theoretical analysis to describe the observed results. PMID- 19547523 TI - Manipulating metal-oxide nanowires using counter-propagating optical line tweezers. AB - Semiconducting nanowires, such as ZnO and Si, are used in the fields of nanophotonics and nanoelectronics. Optical tweezers offer the promise of flexible positional control of such particles in a liquid, but so far this has been limited to either manipulation close to the surface, or to axial trapping of nanowires. We show the three-dimensional trapping of ZnO and silica-coated Si nanowires in counter-propagating line tweezers, and demonstrate translational and rotational in-plane manipulation, away from the surfaces. The high-refractive index particles investigated - ZnO wires (n~1.9) with varying lengths up to 20mum and 6-mum-long silica-coated Si wires (n =3.6) - could not be trapped in single beam line traps. Opposite surface charges are used to fix the nanowires to a surface. Full translational and in-plane rotational control of semiconducting nanowires expands the possibilities to position individual wires in complex geometries significantly. PMID- 19547524 TI - Fluorescence microscopy in a microwave cavity. AB - Optical microscopy is a well-established technique that has wide ranging applications for imaging molecular dynamics of biological systems. Typically, these applications rely on external temperature controllers to maintain or change reactions rates of these biological systems. With increasing interest in applying low power microwaves to drive biological and chemical reactions, we have combined optical and microwave based technologies and developed a fluorescence microscope in a microwave cavity. With this instrument, we have found a means to optically image biological systems inside microwave cavities during the application of microwave pulses. PMID- 19547525 TI - Polarization state measurement of terahertz electromagnetic radiation by three contact photoconductive antenna. AB - We have fabricated a three-contact photoconductive antenna for the polarization sensitive detection of terahertz electromagnetic radiation. Taking into account all three photoconductive signal current components, this three-contact photoconductive antenna can measure the polarization state of pulsed THz radiation at an accuracy comparable to that achieved using the conventional method which employs a set of wire-grid polarizers. The three-contact photoconductive receiver may be useful for polarization-sensitive spectroscopy such as vibrational circular dichroism spectroscopy and ellipsometry in the THz frequency region. PMID- 19547527 TI - Cr(5+):GdVO(4) as a saturable absorber for a diode-pumped Nd:Lu(0.5)Gd(0.5)VO(4) laser. AB - A new Q-switched crystal Cr(5+):GdVO(4) was grown by the Czochralski method for the first time, to our knowledge. Polarized absorption spectra of Cr(5+)+:GdVO(4) were measured at room temperature. The results showed that the crystal has polarized absorption properties, and the absorption band of pi-polarized spectra located at 900 to 1300 nm should be suitable as a passive saturable absorber Q switched laser at about 1 mum. With Cr(5+):GdVO(4) as a saturable absorber, the pulsed laser performance of Nd:Lu(0.5)Gd(0.5)VO(4) at 1.06 mum was demonstrated. The maximum average output power of 122 mW was obtained under a pump power of 3.79 W. The shortest pulse width and largest pulse energy obtained were 361 ns and 0.77 muJ, respectively. To our knowledge, it is the first time the absorption spectra of Cr(5+):GdVO(4) and a pulsed laser with the crystal as the saturable absorber have been obtained. PMID- 19547526 TI - Multifocal multiphoton microscopy based on multianode photomultiplier tubes. AB - Multifocal multiphoton microscopy (MMM) enhances imaging speed by parallelization. It is not well understood why the imaging depth of MMM is significantly shorter than conventional single-focus multiphoton microscopy (SMM). In this report, we show that the need for spatially resolved detectors in MMM results in a system that is more sensitive to the scattering of emission photons with reduced imaging depth. For imaging depths down to twice the scattering mean free path length of emission photons (2xl (s) (em)), the emission point spread function (PSF(em)) is found to consist of a narrow, diffraction limited distribution from ballistic emission photons and a broad, relatively low amplitude distribution from scattered photons. Since the scattered photon distribution is approximately 100 times wider than that of the unscattered photons at 2xl (s) (em), image contrast and depth are degraded without compromising resolution. To overcome the imaging depth limitation of MMM, we present a new design that replaces CCD cameras with multi-anode photomultiplier tubes (MAPMTs) allowing more efficient collection of scattered emission photons. We demonstrate that MAPMT-based MMM has imaging depth comparable to SMM with equivalent sensitivity by imaging tissue phantoms, ex vivo human skin specimens based on endogenous fluorophores, and green fluorescent protein (GFP) expressing neurons in mouse brain slices. PMID- 19547528 TI - In-plane switching dual-frequency liquid crystal cell. AB - The electro-optical responses of the in-plane switching (IPS) dual-frequency liquid crystal (LC) cell operated with the amplitude-modulation method and the frequency-modulation method were investigated. The obtained results reveal that the electric torque exerted to the LCs and the strong anchoring energy produced from the rubbed polyimide dominate the reorientation of the LCs. With the frequency-modulation method, the generated electric torque combined with the strong surface anchoring energy give the cell a very short fall time, which is independent of the applied frequency. A new waveform composed of the amplitude modulation and the frequency modulation of the supplied voltage-pulse to achieve a fast responding IPS LC cell is proposed. The obtained response time is much less than that of the conventional amplitude-modulation method. PMID- 19547529 TI - Ultrafast-laser inscription of a three dimensional fan-out device for multicore fiber coupling applications. AB - A fan-out device has been fabricated using ultrafast-laser waveguide-inscription that enables each core of a multicore optical fiber (MCF) to be addressed by a single mode fiber held in a fiber V-groove array (FVA). By utilizing the unique three-dimensional fabrication capability of this technique we demonstrate coupling between an FVA consisting of a one-dimensional array of fibers and an MCF consisting of a two-dimensional array of cores. When coupled to all cores of the MCF simultaneously, the average insertion loss per core was 5.0 dB in the 1.55 mum spectral region. Furthermore, the fan-out exhibited low cross-talk and low polarization dependent loss. PMID- 19547530 TI - Unusual double four-lobe textures generated by the motion of carbon nanotubes in a nematic liquid crystal. AB - Unusual double four-lobe nematic liquid crystal (LC) textures were observed in the carbon nanotube (CNT)-doped nematic LC under electric field. Through the electro-optical studies in a wide range of vertical electric fields in the direction of the long axis of the LC molecules, it was realized that the double four-lobe nematic LC textures were formed in the range of 120 to 160 V(rms) at 1 Hz. The formation of these unusual double four-lobe nematic LC textures could originate from the electric field-induced movement of CNTs and the subsequently frustrated reorientation of LCs. PMID- 19547531 TI - Efficient terahertz generation by optical rectification at 1035 nm. AB - We demonstrate efficient generation of THz pulses by optical rectification of 1.03 um wavelength laser pulses in LiNbO3 using tilted pulse front excitation for velocity matching between the optical and THz fields. Pulse energies of 100 nJ with a spectral bandwidth of up to 2.5 THz were obtained at a pump energy of 400 uJ and 300 fs pulse duration. This conversion efficiency of 2.5x10(-4) was an order of magnitude higher than that obtained with collinear optical recitification in GaP, and far higher still than that measured using ZnTe in an optimized geometry. Using a simple model we demonstrate that two- and three photon absorption strongly limit the THz generation efficiency at high pump fluences in ZnTe and GaP. PMID- 19547532 TI - Wave scattering and splitting by magnetic metamaterials. AB - We study experimentally propagation of electromagnetic waves through a slab of uniaxial magnetic metamaterial. We observe a range of novel phenomena including partial focusing and splitting into multiple transmitted beams.We demonstrate that while some of these experimentally observed effects can be described within the approximation of an effective medium, a deeper understanding of the experimental results requires a rigorous study of internal eigenmodes of the lattice of resonators. PMID- 19547533 TI - Observation of wavelength and multiple bistabilities in 850nm Vertical-Cavity Semiconductor Optical Amplifiers (VCSOAs). AB - Both wavelength bistability (WB) and multiple bistability (MB) were predicted theoretically by Adams' model in resonant optical amplifiers. We report here, for the first time to our knowledge, their experimental observation in 850nm Vertical Cavity Semiconductor Optical Amplifiers (VCSOA). Clockwise hysteresis of WB is observed at constant input power while the input wavelength is swept across the gain window. MB is observed at a fixed operation wavelength biased on the long wavelength side of the two separated Polarization-Dependent Gain (PDG) windows of the VCSOA by sweeping the optical input. The polarization of the input is set to a fixed angle with respect to the two intrinsic principal axes of the VCSOA. Two MB levels were experimentally observed at 160muW and 320muW, respectively. These observations are in good agreement with theoretical prediction by Adams' model and may lead to multi-valued optical information manipulation. PMID- 19547534 TI - Coherent combining of spectrally broadened fiber lasers. AB - We demonstrate that fiber lasers spectrally broadened by cross mode coupling can be coherently combined with high efficiency. The spectral broadening that it induces suppresses stimulated Brillouin scattering. Using long cavity length lasers, > 800 m, we induce spectral broadening of > 50 GHz and show mode by mode coherence in the output of four intracavity coupled fiber lasers. PMID- 19547535 TI - Hybrid-effect transmission enhancement induced by oblique illumination in nano ridge waveguide. AB - In a nano-ridge waveguide under oblique illumination, we demonstrated transmission enhancement resulting from a hybrid effect between propagation modes and surface plasmon wave. The measured near-field intensity with 44-degree illumination was 1.6 times higher than that illuminated with normal incident light. Consequently, a wedge-shaped fiber probe was proposed to serve as a compact near-field light source. PMID- 19547536 TI - Post-detection nonlinear distortion for efficient MLSD in optical links. AB - In this paper, we investigate the use of nonlinear distortion of the electrical post-detection signal in order to design simple, yet very effective, maximum likelihood sequence detection (MLSD) receivers for optical communications with direct photo-detection. This distortion enables the use of standard Euclidean branch metrics in the Viterbi algorithm which implements MLSD. Our results suggest that the nonlinear characteristic can be optimized with respect to the uncompensated chromatic dispersion and other relevant system parameters, such as the extinction ratio. The proposed schemes with optimized distortion exhibit the same performance of more sophisticated MLSD schemes, still guaranteeing more efficient Viterbi algorithm implementation. PMID- 19547537 TI - Gigahertz optical spin transceiver. AB - We present a time-resolved optical technique to measure electron spin dynamics with GHz dynamical bandwidth, transform-limited spectral selectivity, and phase sensitive (lock-in) detection. Use of a continuous-wave (CW) laser and fast optical bridge enables greatly improved signal-to-noise characteristics compared to traditional optical sampling (pump-probe) techniques. We demonstrate the technique with a measurement of GHz-spin precession in n-GaAs. This approach may be applicable to other physical systems where stroboscopic techniques cannot be used because of either noise or spectral limitations. PMID- 19547538 TI - On-line beam diagnostic method for high energy laser with large beam profile. AB - A novel on-line beam diagnostic method for continuous-wave high energy laser (HEL) is presented. The system based on this method is mainly consisted of a scanning circular reflector and a photodetector array disposed spatially. Laser beam passes through the system except a little part of whole beam is sampled and reflected into the detector array by the circular reflector. Through the arithmetic of spatial mapping and image restoration with the output signal of detector array, the spatial-temporal distribution parameters of the laser beam are obtained. The HEL beam of several hundred millimeters in diameter can be on line measured with spatial resolution of 2 mm and temporal resolution of 30~50ms. PMID- 19547539 TI - Band gap characterization and slow light effects in one dimensional photonic crystals based on silicon slot-waveguides. AB - We investigate the photonic properties of one dimensional photonic crystals realized on Silicon On Insulator channel slot-waveguide to engineer slow light effects. Various geometries of the photonic pattern have been characterized and their photonic band-gap structure analyzed. The optimal geometry has been further used to realize a coupled resonator optical waveguide (CROW). A first optimization of these CROW devices shows a group velocity of more than c/10 at 1.55 mum. Full three dimensional calculations based on the planar wave expansion method have been used to compute the band diagram while full three dimensional calculations based on finite difference time domain methods have been used to study light propagation. PMID- 19547540 TI - Beam quality improvement of laser diode array by using off-axis external cavity. AB - A novel off-axis external cavity is designed for laser diode array to improve the beam quality. In this external cavity, a circle aperture with variable size is used as a spatial filter. The diameter of aperture is optimized to 1.2mm and the off-axis angle of external cavity is optimized at 2.6 deg. In the optimal case, the beam parameter product (BPP) of laser diode array is reduced to 121 mm.mrad from 1050 mm.mrad with external cavity optical efficiency of 81%. PMID- 19547541 TI - Fourier-transform terahertz near-field imaging of one-dimensional slit arrays: mapping of electric-field-, magnetic-field-, and Poynting vectors. AB - We present 2D measurements of the full THz electric field behind a sample consisting of multiple slits in a metal foil. Our measurements, which have a sub wavelength spatial, and a sub-period temporal resolution, reveal electric field lines, electric field vortices and saddle points. From our measurements we are able to reconstruct the magnetic field and, finally, the position and time dependent Poynting vector which shows the flow of energy behind the sample. Our results show that it is possible to study the flow of light near sub-wavelength plasmonic structures such as slit-arrays and, by implication, other metamaterial samples. PMID- 19547542 TI - Generating small-scale structures from large-scale ones via optical near-field interactions. AB - Optical near-fields, which appear in the vicinity of structures when irradiated with light, exhibit a hierarchical nature, meaning that the degree of localization of optical near-fields at a given point is related to the scale of the structure involved in this process. Therefore, if we could make optically induced fabrication processes selectively localized in the near-field region, we could generate a smaller-scale structure even from a larger-scale one via optical near-field interactions. We demonstrate the theoretical basis of this with an angular spectrum analysis of optical near-fields. We also experimentally demonstrate such principles by using ZnO nanoneedles fabricated through metal organic vapor phase epitaxy (MOVPE) followed by a photo-induced MOVPE procedure where smaller-scale generated structures were clearly observed with the help of light irradiation. We also observed that the generated fine structures followed a power-law distribution, indicating that fractal structures emerged via optical near-field interactions. PMID- 19547543 TI - Si-CMOS-compatible lift-off fabrication of low-loss planar chalcogenide waveguides. AB - We demonstrate, for the first time to the best of our knowledge, low-loss, Si CMOS-compatible fabrication of single-mode chalcogenide strip waveguides. As a novel route of chalcogenide glass film patterning, lift-off allows several benefits: leverage with Si-CMOS process compatibility; ability to fabricate single-mode waveguides with core sizes down to submicron range; and reduced sidewall roughness. High-index-contrast Ge(23)Sb(7)S(70) strip waveguides have been fabricated using lift-off with excellent uniformity of loss propagation and the lowest loss figure of reported to date. We also show that small core Ge(23)Sb(7)S(70) rib waveguides can be fabricated via lift-off as well, with loss figures lower than 0.5 dB/cm. Additionally, we find through waveguide modal analysis that although overall transmission loss is low, the predominant source of this loss comes from scattering at the sidewalls. PMID- 19547544 TI - Transient fields in the input coupling region of optical single-mode waveguides. AB - We investigate numerically the optical field in the region immediately behind the input facets of dielectric step-index single-mode slab and fiber waveguides. Visualization of the intensity distributions gives insight into the formation of the fundamental mode and of radiation modes. For a more quantitative characterization we determine the amount of optical power and mode purity of the field in core vicinity as a function of propagation distance. The investigation assists in designing and optimizing waveguides being employed as modal filters, e.g. for astronomical interferometers. PMID- 19547545 TI - Optical wave properties of nano-particle chains coupled with a metal surface. AB - Optical phenomena supported by ordered and disordered chains of metal nano particles on a metal surface are investigated by considering a particular example of gold nano-bumps on a gold surface. The TWs supported by these structures are analyzed by studying the frequency-wavenumber spectra of the fields excited by localized sources placed near the chain. Periodic nano-bump chains support traveling waves (TWs) that propagate without radiation loss along, and are confined to the region near, the chain. These TWs are slow waves with respect to both space fields and surface plasmon polaritons supported by the metal surface. For nearly resonant nano-bumps, the TWs are well confined and can be excited efficiently by a localized source placed near the chain but the TW propagation length is short. For non-resonant nano-bumps, the TWs have large propagation lengths but are not well confined and are excited less efficiently. The TWs supported by nano-bump chains were shown to have larger propagation lengths than free-standing chains of the same dimension/size and cross-sectional confinement. TWs also are supported by disordered chains and chains with sharp bends. Perturbations in nano-bump positions are shown to reduce the TW propagation length much less significantly than perturbations in their sizes. Transmission through sharp chain bends is much stronger for nearly resonant nano-bumps than for nonresonant ones. In addition to their ability to support TWs, nano-bump chains can be used to manipulate (excite/reflect/refract) SPPs on the metal surface. PMID- 19547546 TI - Opening up optical fibres. AB - A unique optical fibre design is presented in this work: a laterally accessible microstructured optical fibre, in which one of the cladding holes is open to the surrounding environment and the waveguide core exposed over long lengths of fibre. Such a fibre offers the opportunity of real-time chemical sensing and biosensing not previously possible with conventional microstructured optical fibres, as well as the ability to functionalize the core of the fibre without interference from the cladding. The fabrication of such a fibre using PMMA is presented, as well as experimental results demonstrating the use of the fibre as a evanescent wave absorption spectroscopy pH sensor using the indicator Bromothymol Blue. PMID- 19547547 TI - Observation of electromagnetically induced transparency for a squeezed vacuum with the time domain method. AB - A probe light in a squeezed vacuum state was injected into cold 87Rb atoms with an intense control light in a coherent state. A sub-MHz window was created due to electromagnetically induced transparency, and the incident squeezed vacuum could pass through the cold atoms without optical loss, as was successfully monitored using a time-domain homodyne method. PMID- 19547548 TI - Raman frequency shifter for laser pulses shorter than 100 fs. AB - The technique of frequency shifting of sub-100 fs laser pulses was developed. It is based on the stimulated Raman scattering pair of chirped laser pulses with orthogonal polarization. The 50 fs laser pulse at the wavelength of 810 nm was converted to 68 fs Stokes pulse at the wavelength of 1060 nm with energy conversion efficiency of 20%. PMID- 19547549 TI - The dark spots of Arago. AB - We explore the diffraction and propagation of Laguerre- Gaussian beams of varying azimuthal index past a circular obstacle both experimentally and numerically. When the beam and obstacle centers are aligned the famous spot of Arago, which arises for zero azimuthal index, is replaced for non-zero azimuthal indices by a dark spot of Arago, a simple consequence of the conserved phase singularity at the beam center. We explore how the dark spot of Arago behaves as the beam and obstacle centers are progressively misaligned, and find that the central dark spot may break into several dark spots of Arago for higher incident azimuthal index beams. PMID- 19547550 TI - Development of a handheld Raman microspectrometer for clinical dermatologic applications. AB - Although skin is easily accessible to optical methodologies, a portable measurement head is necessary to allow ready spectroscopic interrogation of all anatomic locations. However, most conventional Raman microspectrometers and even dermatologic-specific Raman systems are fixed systems ill-suited to anatomic accessibility. To this end, we have developed a portable Raman microspectrometer system for future dermatologic studies. An in-house-built bench-top system was used to qualify the optical components and design. Based on this system's layout, a handheld microspectrometer was developed for future clinical application. This system produces similar operating characteristics to the bench-top prototype, and is shown to provide clear Raman spectra from skin tissue measured in vivo in clinically-feasible integration times. PMID- 19547551 TI - Analysis of the optimum optical incident angle for an imaging acousto-optic tunable filter. AB - The selection of the incident polar angle is very important in the entire design of the noncollinear acousto-optic tunable filter (AOTF). The authors discussed how the factors, including tuning range of wavelength, the acoustic frequency, the acousto-optic figure of merit, the spectral bandwidth, the spread of filtered beam and the wavelength dependence, influence the selection of the optimum incident polar angle. By an accurate theoretical analysis, a method of selecting the optimum incident angle was presented. The analysis was significant for improving the performance of the imaging AOTF from the visible to the infrared. PMID- 19547552 TI - Performance of 3D integral imaging with position uncertainty. AB - We present the theoretical and simulation results on the analysis of Synthetic Aperture Integral Imaging (SAII) technique and its sensitivity to pickup position uncertainty. SAII is a passive three dimensional imaging technique based on multiple image acquisitions with different perspective of the scene under incoherent or natural illumination. In practical SAII applications, there is always an uncertainty associated with the position at which each sensor captures the elemental image. We present a theoretical analysis that quantifies image degradation in terms of Mean Square Error (MSE) metric. Simulation results are also presented to identify the parameters affecting the reconstruction degradation and to confirm the analysis. We show that in SAII with a given uncertainty in the sensor locations, the high spatial frequency content of the 3D reconstructed images are most degraded. We also show an inverse relationship between the reconstruction distance and degradation metric. To the best of our knowledge, this is the first time that the effects of sensor position uncertainty on 3D computational reconstruction in synthetic aperture integral imaging systems have been quantitatively analyzed. PMID- 19547553 TI - Computational model for operation of 2 mum co-doped Tm,Ho solid state lasers. AB - A computational model for operation of co-doped Tm,Ho solid-state lasers is developed coupling (i) 8-level rate equations with (ii) TEM00 laser beam distribution, and (iii) complex heat dissipation model. Simulations done for Q switched approximately 0.1 J giant pulse generation by Tm,Ho:YLF laser show that approximately 43% of the 785 nm light diode side-pumped energy is directly transformed into the heat inside the crystal, whereas approximately 45% is the spontaneously emitted radiation from (3)F(4), (5)I(7) , (3)H(4) and (3)H(5) levels. In water-cooled operation this radiation is absorbed inside the thermal boundary layer where the heat transfer is dominated by heat conduction. In high power operation the resulting temperature increase is shown to lead to (i) significant decrease in giant pulse energy and (ii) thermal lensing. PMID- 19547554 TI - External power-enhancement cavity versus intracavity frequency doubling of Ti:sapphire lasers using BIBO. AB - We report on continuous-wave second harmonic generation of near infrared Ti:sapphire lasers using room temperature critically phase-matched, angle-tuned BIBO (bismuth triborate, BiB(3)O(6)) crystals, placed both in an external power enhancement cavity and inside the laser resonator. In the first case we generate 70 mW of single-frequency radiation at 423 nm for 330 mW of input power at 846 nm. For intracavity frequency doubling we achieve 690 mW at 423 nm for 7.3 Watts of the Ti:sapphire laser pump power at 532 nm, representing a conversion efficiency of 9.5% from 532 to 423 nm. These tunable blue-violet systems are particularly attractive for laser cooling and trapping of alkaline-Earth atoms. PMID- 19547555 TI - Artificial neural superposition eye. AB - We propose an ultra-thin imaging system which is based on the neural superposition compound eye of insects. Multiple light sensitive pixels in the footprint of each lenslet of this multi-channel configuration enable the parallel imaging of the individual object points. Together with the digital superposition of related signals this multiple sampling enables advanced functionalities for artificial compound eyes. Using this technique, color imaging and a circumvention for the trade-off between resolution and sensitivity of ultra-compact camera devices have been demonstrated in this article. The optical design and layout of such a system is discussed in detail. Experimental results are shown which indicate the attractiveness of microoptical artificial compound eyes for applications in the field of machine vision, surveillance or automotive imaging. PMID- 19547556 TI - Ultra-compact high order ring resonator filters using submicron silicon photonic wires for on-chip optical interconnects. AB - Ultra-compact 5(th) order ring resonator optical filters based on submicron silicon photonic wires are demonstrated. Out-of-band rejection ratio of 40dB, 1dB flat-top pass band of 310GHz with ripples smaller than 0.4dB, and insertion loss of only (1.8+/-0.5)dB at the center of the pass band are realized simultaneously, all within a footprint of 0.0007mm(2) on a silicon chip. PMID- 19547557 TI - Nonparaxial analysis of vectorial Laguerre-Bessel-Gaussian beams. AB - The concept of vectorial Laguerre-Bessel-Gaussian (LBG) beams is proposed. On the basis of vectorial Rayleigh-Sommerfeld formulas, the analytical formulas for the nonparaxial propagation of vectorial LBG beams are derived and applied to study the nonparaxial propagation properties of vectorial LBG beams. The far field and paraxial approximation are dealt with as special cases of our general results. Some detailed comparisons of the obtained results with the paraxial results are made, which show the propagation of paraxial and nonparaxial LBG beams is all instable in the near field and the f parameter plays the important role in determining the nonparaxiality of vectorial LBG beams. The beam parameter alpha also affects the propagation behavior of nonparaxial LBG beams. Under certain conditions, the obtained results can be reduced to those of the cases for vectorial Laguerre-Gaussian and Bessel Gaussian beams. PMID- 19547558 TI - Ultra-sensitive surface absorption spectroscopy using sub-wavelength diameter optical fibers. AB - The guided modes of sub-wavelength diameter air-clad optical fibers exhibit a pronounced evanescent field. The absorption of particles on the fiber surface is therefore readily detected via the fiber transmission. We show that the resulting absorption for a given surface coverage can be orders of magnitude higher than for conventional surface spectroscopy. As a demonstration, we present measurements on sub-monolayers of 3,4,9,10-perylene-tetracarboxylic dianhydride (PTCDA) molecules at ambient conditions, revealing the agglomeration dynamics on a second to minutes timescale. PMID- 19547559 TI - Theory of resonantly enhanced near-field imaging. AB - We investigate the fundamental issues of power transfer and far-field retrieval of subwavelength information in resonantly enhanced near-field imaging systems. It is found that high-quality resonance of the imaging system, such as that provided by dielectric resonators, can drastically enhance the power transfer from the object to the detector or the working distance. The optimal power transfer condition is shown to be the same as the critical coupling condition for resonators. The combination of a dielectric planar resonator with a solid immersion lens is proposed to project resonantly enhanced near-field spatial frequency components into the far field with the same resolution limit as that for solid immersion microscopy, but with much improved signal power throughput or working distance for resonant spatial frequencies. PMID- 19547560 TI - Generalized phase contrast matched to Gaussian illumination. AB - We show that the generalized phase contrast method (GPC) can be used as a versatile tool for shaping an incident Gaussian illumination into arbitrary lateral beam profiles. For illustration, we use GPC in an energy-efficient phase only implementation of various apertures that do not block light but instead effectively redirect the available photons from a bell-shaped light distribution. GPC-based generation of lateral beam profiles can thus be achieved using a simplified optical implementation as it eliminates the need for a potentially lossy initial beam shaping. The required binary phase input is simple to fabricate for static applications and can be easily reconfigured up to device frame refresh rates for dynamic applications. PMID- 19547561 TI - Frequency-selective self-trapping and supercontinuum generation in arrays of coupled nonlinear waveguides. AB - We study spatiotemporal dynamics of soliton-induced twooctave- broad supercontinuum generated by fs pulses in an array of coupled nonlinear waveguides. We show that after fission of the input pulse into several fundamental solitons, red and blue-shifted nonsolitonic radiation, as well as solitons with lower intensity, spread away in transverse direction, while the most intense spikes self-trap into spatiotemporal discrete solitons. PMID- 19547562 TI - Optical trapping of nonspherical particles in the T-matrix formalism. AB - The theory of the trapping of nonspherical particles in the focal region of a high-numerical-aperture optical system is formulated in the framework of the transition matrix approach. Both the case of an unaberrated lens and the case of an aberrated one are considered. The theory is applied to single latex spheres of various sizes and, when the results are compared with the available experimental data, a fair agreement is attained. The theory is also applied to binary clusters of spheres of latex with a diameter of 220nm in various orientations. Although, in this case we have no experimental data to which our results can be compared, we get useful indications for the trapping of nonspherical particles. In particular, we find substantial agreement with recent results on the trapping of prolate spheroids in aberrated gaussian fields [S. H. Simpson and S. Hanna, J. Opt.Soc. Am. A 24, 430 (2007)]. PMID- 19547563 TI - Evidence for inhibited diffraction of light propagating through nanolaminate metallodielectric material. AB - We report the fabrication and characterization of laterally continuous silver layers alternated with glassy amorphous polycarbonate films with the thickness of each layer much less than the wavelength. Such films exhibit physical phenomena associated with the coupled plasmon resonances. We have characterized light propagation through the resulting metal-dielectric (MD) periodic structures using collection mode Near Field Scanning Optical Microscopy (NSOM). In agreement with published theoretical models, our experiments provide evidence that diffraction can be inhibited for light propagating through metallodielectric nanolaminate. PMID- 19547564 TI - Thickness dependence of optical second harmonic generation in collagen fibrils. AB - Simultaneous backward and forward second harmonic generations from isolated type I collagen matrix are observed. Optical interference behaviors of these nonlinear optical signals are studied with accurately determined fibril thickness by an atomic force microscope. The nonlinear emission directions are strongly dependent on the coherent interaction within and between collagen fibrils. A linear relationship is obtained to estimate collagen fibril thickness with nanometer precision noninvasively by evaluating the forward/backward second harmonic generation ratio. PMID- 19547565 TI - Refractometric sensor based on whispering-gallery modes of thin capillarie. AB - Whispering-gallery modes resonances of submicron wall thickness capillaries exhibit very large wavelength shifts as a function of the refractive index of the medium that fills the inside. The sensitivity to refractive index changes is larger than in other optical microcavities as microspheres, microdisks and microrings. The outer surface where total internal reflection takes place remains always in air, enabling the measure of refractive index values higher than the refractive index of the capillary material. The fabrication of capillaries with submicron wall thickness has required the development of a specific technique. A refractometer with a response higher than 390 nm per refractive index unit is demonstrated. These sensors are readily compatible with microfluidic systems. PMID- 19547566 TI - Plasmon resonance-based optical trapping of single and multiple Au nanoparticles. AB - The plasmon resonance-based optical trapping (PREBOT) method is used to achieve stable trapping of metallic nanoparticles of different shapes and composition, including Au bipyramids and Au/Ag core/shell nanorods. In all cases the longitudinal plasmon mode of these anisotropic particles is used to enhance the gradient force of an optical trap, thereby increasing the strength of the trap potential. Specifically, the trapping laser is slightly detuned to the long wavelength side of the longitudinal plasmon resonance where the sign of the real component of the polarizability leads to an attractive gradient force. A second (femtosecond pulsed) laser is used to excite two-photon fluorescence for detection of the trapped nanoparticles. Two-photon fluorescence time trajectories are recorded for up to 20 minutes for single and multiple particles in the trap. In the latter case, a stepwise increase reflects sequential loading of single Au bipyramids. The nonlinearity of the amplitude and noise with step number are interpreted as arising from interactions or enhanced local fields amongst the trapped particles and fluctuations in the arrangements thereof. PMID- 19547567 TI - Response function calculation and sensitivity comparison analysis of various bimorph deformable mirrors. AB - Analysis of the response function for various possible bimorph-type deformable mirrors is presented by numerical calculation. Using this response function, sensitivity comparison of a five-layer bimorph with two-and three-layer structures is carried out. It is shown that the displacement of a bimorph deformable mirror surface reduces when the number of layers increases, and the displacement is closely related to the distance of the control layer from the median plane. The farther electrode contributes the larger deformation on the surface. Furthermore, for a fixed position of the control layer, the displacement is directly proportional to the loading voltage and inversely proportional to the square of the thickness of the PZT layer, and it increases about 1.4 times when the area of the electrode doubles. PMID- 19547568 TI - Image quality enhancement in 3D computational integral imaging by use of interpolation methods. AB - In this paper, we propose a computational integral imaging reconstruction (CIIR) method by use of image interpolation algorithms to improve the visual quality of 3D reconstructed images. We investigate the characteristics of the conventional CIIR method along the distance between lenslet and objects. What we observe is that the visual quality of reconstructed images is periodically degraded. The experimentally observed period is half size of the elemental image. To remedy this problem, we focus on the interpolation methods in computational integral imaging. Several interpolation methods are applied to the conventional CIIR method and their performances are analyzed. To objectively evaluate the proposed CIIR method, we introduce an experimental framework for the computational pickup process and the CIIR process using a Gaussian function. We also carry out experiments on real objects to subjectively evaluate the proposed method. Experimental results indicate that our method outperforms the conventional CIIR method. In addition, our method reduces the grid noise that the conventional CIIR method suffers from. PMID- 19547569 TI - Differential imaging in coherent anti-Stokes Raman scattering microscopy II: a filter-assisted Laguerre-Gaussian beam detection scheme. AB - A new differential imaging technique to obtain contrast improvement in coherent anti-Stokes Raman scattering (CARS) microscopy is proposed through a spatial spiral phase modulation in the collected signal of CARS microscopy. A spiral phase mask makes the CARS signal from the bulk material to be distributed in a ring centered at the detecting pinhole of a confocal microscope resulting in a weak detected CARS signal from a bulk material. When tiny scatters are included in the focal volume of a CARS setup, the ring-shaped distribution of CARS field is distorted, leading to an increase in the detected signal through the pinhole. The sensitivity and the size selectivity of this proposed technique is studied with varying the particle size, and it is found that this method is to be efficient in edge detection. Simulation results obtained by finite difference time domain (FDTD) methods show that the image contrast is enhanced by many times as it is able to highlight the details of the specimen by suppressing the CARS signal from a bulk or a uniform material. PMID- 19547570 TI - Absorption coefficients of selected explosives and related compounds in the range of 0.1-2.8 THz. AB - We have investigated the absorption spectra of seventeen explosives and related compounds (ERCs) by using terahertz time-domain spectroscopy in the 0.1-2.8 THz region. Most of these substances show characteristic absorption features in this frequency range. The measured absorption coefficients of these ERCs form a database, which is of great importance for biochemical, defense and security related applications. PMID- 19547571 TI - Single beam mapping of nonlinear phase shift profiles in planar waveguides with an embedded mirror. AB - We demonstrate a technique for a single shot mapping of nonlinear phase shift profiles in spatial solitons that are formed during short pulse propagation through one-dimensional slab AlGaAs waveguides, in the presence of a focusing Kerr nonlinearity. The technique uses a single beam and relies on the introduction of a lithographically etched reflective planar mirror surface positioned in proximity to the beam's input position. Using this setup we demonstrate nonlinearity-induced sharp lateral phase variations for certain initial conditions, and creation of higher spatial harmonics when the beam is in close proximity to the mirror. PMID- 19547572 TI - Chemically-selective imaging of brain structures with CARS microscopy. AB - We demonstrate the use of coherent anti-Stokes Raman scattering (CARS) microscopy to image brain structure and pathology ex vivo. Although non-invasive clinical brain imaging with CT, MRI and PET has transformed the diagnosis of neurologic disease, definitive pre-operative distinction of neoplastic and benign pathologies remains elusive. Definitive diagnosis still requires brain biopsy in a significant number of cases. CARS microscopy, a nonlinear, vibrationally sensitive technique, is capable of high-sensitivity chemically-selective three dimensional imaging without exogenous labeling agents. Like MRI, CARS can be tuned to provide a wide variety of possible tissue contrasts, but with sub cellular spatial resolution and near real time temporal resolution. These attributes make CARS an ideal technique for fast, minimally invasive, non destructive, molecularly specific intraoperative optical diagnosis of brain lesions. This promises significant clinical benefit to neurosurgical patients by providing definitive diagnosis of neoplasia prior to tissue biopsy or resection. CARS imaging can augment the diagnostic accuracy of traditional frozen section histopathology in needle biopsy and dynamically define the margins of tumor resection during brain surgery. This report illustrates the feasibility of in vivo CARS vibrational histology as a clinical tool for neuropathological diagnosis by demonstrating the use of CARS microscopy in identifying normal brain structures and primary glioma in fresh unfixed and unstained ex vivo brain tissue. PMID- 19547573 TI - Fabrication of plastic microlens arrays using hybrid extrusion rolling embossing with a metallic cylinder mold fabricated using dry film resist. AB - This paper reports a novel and effective method to fabricate microlens arrays on polycarbonate films by hybrid extrusion rolling embossing. The metallic cylinder mold bearing an array of micro-holes is fabricated using photolithography with dry film resist. During the extrusion rolling embossing process, the extruded PC film is immediately pressed against the surface of the roller mold. Under the influence of the rolling pressure and surface tension, an array of convex microlenses is formed. The uniformity and optical properties have been verified. An efficient continuous mass production technique has been demonstrated. PMID- 19547574 TI - Resonance hybridization in double split-ring resonator metamaterials. AB - We introduce a plasmon hybridization picture to understand the optical properties of double split-ring resonator metamaterials. The analysis is based on the calculated reflectance spectra from a finite-integration time-domain algorithm. Field distributions of the double split-ring resonators at the resonant frequencies confirm the results from the plasmon hybridization analysis. We demonstrate that the plasmon hybridization is a simple and powerful tool for understanding and designing metamaterials in the near infrared and visible regime. PMID- 19547575 TI - Ultrafast all-optical Nth-order differentiator and simultaneous repetition-rate multiplier of periodic pulse train. AB - The letter presents a technique for Nth-order differentiation of periodic pulse train, which can simultaneously multiply the input repetition rate. This approach uses a single linearly chirped apodized fiber Bragg grating, which grating profile is designed to map the spectral response of the Nth-order differentiator, and the chirp introduces a dispersion that, besides space-to-frequency mapping, it also causes a temporal Talbot effect. PMID- 19547576 TI - Photo-assisted electrical gating in a two-terminal device based on vanadium dioxide thin film. AB - For electrical devices based on vanadium dioxide thin film, various methods have been implemented on the electrical gating of the devices. In this paper, a photo assisted electrical gating in a two-terminal device is demonstrated based on vanadium dioxide thin film, instead of a three-terminal device with a gate terminal, by illuminating infrared light directly onto the film. Based on the light-induced phase transition, the threshold voltage of the device, in which an abrupt current jump take places, was theoretically anticipated to be controlled (electrically gated) by adjusting the light intensity. Finally, the prediction was experimentally verified. PMID- 19547577 TI - Direct measurement of the Atom number in a Bose condensate. AB - We report on directly measuring the atom number in a Bose- Einstein condensate by the method of optical pumping. Only the branching ratio of the spontaneous decay in the system and the absorption energy of a probe laser beam are required to determine the atom number. The measured absorption energy is not affected by the measurement condition such as the intensity, detuning, and polarization of the probe beam, the magnetic field, etc. We have shown that atom numbers as low as a few thousands can be measured. The atom number is an important parameter in the studies of Bose condensates and its accuracy is greatly improved by this sensitive and robust method. PMID- 19547578 TI - Low-saturation-energy-driven ultrafast all-optical switching operation in (CdS/ZnSe)/BeTe intersubband transition. AB - The authors report their latest results on II-VI intersubband all-optical switches in which the 10 dB absorption saturation energy is lowered to ~2.0-2.2 pJ for 1.55-1.58 mum by decreasing the thickness of the active layer and increasing the refractive index difference between the core layer and the cladding layers in waveguides. Such low saturation energies greatly improve the switching performance. <7 pJ pump energy at 1520 nm is sufficient for realizing 10 dB switching operation at 1566 nm (switching energy: ~0.7 pJ/dB). To the best of our knowledge, these switching energy and saturation energy values are the lowest reported ones for such ultrafast intersubband all-optical switches at telecommunication wavelengths. PMID- 19547579 TI - Fluorescence near metal tips: The roles of energy transfer and surface plasmon polaritons. AB - We simulate the remarkable changes that occur to the decay rates of a fluorescent molecule as a conical metal tip approaches. A new and simple model is developed to reveal and quantify which decay channels are responsible. Our analysis, which is independent of the method of molecular excitation, shows some universal characteristics. As the tip-apex enters the molecule's near-field, the creation of surface plasmon polaritons can become extraordinarily efficient, leading to an increase in the nonradiative rate and, by proportional radiative-damping, in the radiative rate. Ehancements reaching 3 orders of magnitude have been found, which can improve the apparent brightness of a molecule. At distances less than ~5nm, short-ranged energy transfer to the nano-scale apex quickly becomes dominant and is entirely nonradiative. PMID- 19547580 TI - Tunable diffraction and self-defocusing in liquid-filled photonic crystal fibers. AB - We suggest and demonstrate a novel platform for the study of tunable nonlinear light propagation in two-dimensional discrete systems, based on photonic crystal fibers filled with high index nonlinear liquids. Using the infiltrated cladding region of a photonic crystal fiber as a nonlinear waveguide array, we experimentally demonstrate highly tunable beam diffraction and thermal self defocusing, and realize a compact all-optical power limiter based on a tunable nonlinear response. PMID- 19547581 TI - Effect of heat accumulation on the dynamic range of a gold nanorod doped polymer nanocomposite for optical laser writing and patterning. AB - Even though gold nanorod doped dielectrics have been widely used for optical laser writing and patterning there has been no attempt to study the dynamic range of these nanocomposites, let alone exploring ways to improve this property. Here we study the dynamic range of a gold nanorod doped polyvinyl alcohol film for various laser spot sizes at two different laser pulse repetition rates and show that when a high repetition rate laser source is employed the dynamic range of the nanocomposite is severely limited due to accumulative heating inside the focal volume. This problem could be solved by silica-coating the nanorods inside the polymer matrix. This method does not compromise the high repetition rate of the laser writing source and yet retains the attractive flexible properties of the polymer matrix. The silica-coated gold nanorod doped polymer nanocomposite could be an attractive medium for future high-speed, high repetition rate pulsed laser writing and patterning applications. PMID- 19547582 TI - Self referenced Yb-fiber-laser frequency comb using a dispersion micromanaged tapered holey fiber. AB - We demonstrate a fully stabilized frequency comb in the 1mum spectral region based on an Yb-fiber oscillator and a cladding pumped chirped pulse Yb-fiber amplifier whose output is spectrally broadened in a dispersion micromanaged holey fiber. The dispersion micromanaged fiber is used to generate efficient, low noise spectral components at 523nm which are heterodyned with the second harmonic of the amplifier output for standard f-to-2f self-referenced carrier envelope offset frequency detection. For comb stabilization we phase-lock this offset frequency and the oscillator repetition frequency simultaneously to an RF reference by feedback controlling the oscillator pump diode current and the driving voltage of an intracavity piezo-electric fiber stretcher respectively. PMID- 19547583 TI - Simultaneous noise and distortion reduction of a broadband optical feedforward transmitter for multi-service operation in radio-over-fiber systems. AB - The broadband optical feedforward transmitter with uncooled and unisolated distributed-feedback laser diodes (DFB LDs) is developed for a radio-over-fiber system. Although we use DFB LDs for digital communications, the feedforward compensation method can significantly suppress the intermodulation distortions and background noise. For the wide frequency range from 2.05 to 2.60 GHz (550 MHz), the third-order intermodulation distortion (IMD3) is suppressed by more than 10 dB. We also analyze the variation of IMD3 and noise for the bias current of LDs. With the linearization technique, the maximum IMD3 suppression and spurious-free dynamic range enhancement are 21.3 dB and 7.11 dB, respectively, at 2.3 GHz. PMID- 19547584 TI - Figures of merit for 2D surface plasmon waveguides and application to metal stripes. AB - Three figures of merit, useful as quality measures for 2D surface plasmon waveguides, are discussed and applied to help trade-off mode confinement against attenuation for the symmetric mode propagating along metal stripes. Different stripe geometries are considered, and Au, Ag and Al are compared as the stripe metal over the wavelength range from 200 to 2000 nm. Depending on which figure of merit is used, and on how mode confinement is measured, different preferred designs emerge. For instance, given a mode area, narrow thick stripes are better than wide thin ones, but given a distance from the light line, the opposite is true. Each of the metals analyzed show wavelength regions where their performance is best. The figures of merit are generally applicable and should be useful to help compare, assess and optimize designs in other 2D surface plasmon waveguides or in other absorbing waveguides. PMID- 19547585 TI - Optical feedback effect in DFB lasers for remote reflectivity detecting. AB - A new approach for remote reflectivity detecting based on optical feedback effect in distributed feedback (DFB) lasers is presented. A linear dependent relationship between the reflectivity of external target and the signal modulation depth is obtained. The experimental results show a good agreement with the theoretical analysis and the simulation, and indicate that the active sensing based on optical feedback effect in DFB laser is an effective approach for reflectivity detecting. With the advantage of simple and compact structure, this application can easily enhance the development of a new generation of active sensor. PMID- 19547586 TI - Polarization degree analysis of objective spectrum in polarization differential stellar coronagraph. AB - It is shown that the degree of polarization analysis is useful to find objective spectra of exoplanets immersed in noisy stellar spectra. We report the laboratory experiment of polarization differential objective spectroscopy with a four quadrant polarization mask coronagraph, where partially polarized planetary signal is expected to be discerned from unpolarized stellar noise. The detection of the planet signal is impeded by the stellar noise remained after subtracting mutually orthogonally polarized components of light. We distinguish clearly the planetary spectrum by use of the degree of polarization. We also show the refinement of the spectrum of the planet model. PMID- 19547587 TI - Subsurface damage in precision ground ULE(R) and Zerodur(R) surfaces. AB - The total process cycle time for large ULE((R)) and Zerodur((R))optics can be improved using a precise and rapid grinding process, with low levels of surface waviness and subsurface damage. In this paper, the amounts of defects beneath ULE((R)) and Zerodur((R) )surfaces ground using a selected grinding mode were compared. The grinding response was characterised by measuring: surface roughness, surface profile and subsurface damage. The observed subsurface damage can be separated into two distinct depth zones, which are: 'process' and 'machine dynamics' related. PMID- 19547588 TI - Interferometric resolution improvement for confocal microscopes. AB - We present a method for increasing the lateral resolution and detection efficiency of scanning fluorescence microscopes by adding an interferometer with partial image inversion to the detection pathway. We show that the resulting detection transfer function is essentially the absolute square of the system's amplitude transfer function enlarged to twice its spatial frequency range. Simulations for a confocal system yield a lateral FWHM resolution of 168 nm (135 nm after image subtraction) as compared to 218 nm for confocal detection without an interferometer. Furthermore we demonstrate how this method is suitable for extended focus imaging. Here simulations for Bessel beam excitation and interferometric detection yield a resolution of 146 nm (116 nm after image subtraction) as compared to 199 nm for integrating detection without an interferometer. PMID- 19547589 TI - A tomographic approach to inverse mie particle characterization from scattered light. AB - The problem of computing the internal electromagnetic field of a homogeneous sphere from the observation of its scattered light field is explored. Using empirical observations it shown that, to good approximation for low contrast objects, there is a simple Fourier relationship between a component of the internal E-field and the scattered light in a preferred plane. Based on this relationship an empirical algorithm is proposed to construct a spherically symmetric particle of approximately the same diameter as the original, homogeneous, one. The size parameter (ka) of this particle is then estimated and shown to be nearly identical to that of the original particle. The size parameter can then be combined with the integrated power of the scatter in the preferred plane to estimate refractive index. The estimated values are shown to be accurate in the presence of moderate noise for a class of size parameters. PMID- 19547590 TI - A high sensitive fiber SERS probe based on silver nanorod arrays. AB - A portable fiber SERS probe has been developed based on Ag nanorod array fabricated by oblique angle deposition. The incoming laser beam was designed to focus onto the Raman substrate at 45 degrees incident angle in order to maximize surface enhanced Raman scattering signal. With a fiber Raman system, a detection sensitivity of 10(-17) moles for trans-1, 2- bis(4-pyridyl)ethane molecules has been demonstrated. This Raman probe can also be used for in situ measurement for samples in aqueous solution. Such a fiber probe has great potential as a portable and remote sensor for on-site biological or chemical detection. PMID- 19547591 TI - Efficient visible light generation by mixing of a solid-state laser and a tapered diode laser. AB - A generic approach to efficient visible light generation based on singly-resonant sum-frequency mixing of an external-cavity tapered diode laser and a diode pumped solid-state laser is presented. The principle is exemplified by generation of more than 300 mW of 488 nm coherent blue light by mixing of a 950 mW beam from an external-cavity 765 nm tapered diode laser with the intra-cavity field of a diode pumped, high finesse 1342 nm solid-state laser using periodically poled KTP as the nonlinear medium. Using this approach, a conversion efficiency of more than 30 % of the 765 nm beam was obtained. PMID- 19547592 TI - Parametric frequency conversion of short optical pulses controlled by a CW background. AB - We predict that parametric sum-frequency generation of an ultra-short pulse may result from the mixing of an ultra-short optical pulse with a quasi-continuous wave control. We analytically show that the intensity, time duration and group velocity of the generated idler pulse may be controlled in a stable manner by adjusting the intensity level of the background pump. PMID- 19547593 TI - Increasing the bit rate in OCDMA systems using pulse position modulation techniques. AB - We have experimentally demonstrated two novel pulse position modulation techniques, namely Double Pulse Position Modulation (2-PPM) and Differential Pulse Position Modulation (DPPM) in Time-Wavelength OCDMA systems that will operate at a higher bit rate compared to traditional OOK-OCDMA systems with the same bandwidth. With 2-PPM technique, the number of active users will be more than DPPM while their bit rate is almost the same. Both techniques provide variable quality of service in OCDMA networks. PMID- 19547594 TI - Size dependent multiphoton absorption and refraction of CdSe nanoparticles. AB - We report a systematic investigation on nonlinear optical properties of CdSe nanoparticles that are smaller as well as larger than the Bohr radius. Multiphoton absorption and nonlinear refraction properties of CdSe nanoparticles observed with 800nm wavelength and 110femtosecond Ti:Sapphire laser are presented. These nonlinear optical studies were undertaken by performing open and closed aperture Z-scan measurements. The four different sizes of CdSe nanoparticles investigated are 5nm, 10nm, 25nm and 400nm. Both the quantum dots 5nm, 10nm sizes (taking the literature value of 10.6nm as the Bohr exciton diameter) show four photon absorption (4PA), while the 25nm and 400nm show the three photon absorption (3PA) properties. All four sizes of CdSe nanoparticles show the positive nonlinear refraction (n2). PMID- 19547595 TI - Toward terabyte two-photon 3D disk. AB - 253GB have been recorded in 300 layers inside the volume of one of our two-photon 3D disks. Each layer contains the equivalent of CD layer bit-densities recorded with a 0.5NA objective lens. A new 1.0NA lens with the desirable first order optical properties of long working distance and small diameter, 1.2mm and 4.5mm, and a self-compensating spherical aberration correction mechanism is designed, manufactured and integrated into our single beam two-photon 3D automated recording system. Experimental data obtained with the 1.0NA lens are presented. The resulting bit densities obtained with our new high-performance liquid immersion singlet (LIS) objective lens indicate that our system is capable of full disk recordings from 0.5 to 1 TB within a standard optical disk form factor of 120mm x 1.2mm thick utilizing our very stable and efficient materials. A compact optical head based on our new objective lens capable of TB storage is described. PMID- 19547596 TI - Initial results of in vivo non-invasive cancer imaging in the human breast using near-infrared photoacoustics. AB - Near-infrared photoacoustic images of regions-of-interest in 4 of the 5 cases of patients with symptomatic breasts reveal higher intensity regions which we attribute to vascular distribution associated with cancer. Of the 2 cases presented here, one is especially significant where benign indicators dominate in conventional radiological images, while photoacoustic images reveal vascular features suggestive of malignancy, which is corroborated by histopathology. The results show that photoacoustic imaging may have potential in visualizing certain breast cancers based on intrinsic optical absorption contrast. A future role for the approach could be in supplementing conventional breast imaging to assist detection and/or diagnosis. PMID- 19547597 TI - Estimation of helical angles of myosin and collagen by second harmonic generation imaging microscopy. AB - We performed Second Harmonic Generation (SHG) imaging microscopy of endogeneous myosin-rich and collagen-rich tissues in amphibian and mammals. We determined the relative components of the macroscopic susceptibility tensor chi((2)) from polarization dependence of SHG intensity. The effective orientation angle theta(e) of the harmonophores has been determined for each protein. For myosin we found theta(e) approximately 62 degrees and this value was unchanged during myofibrillogenesis. It was also independent of the animal species (xenopus, dog and human). For collagen we found theta(e) approximately 49 degrees for both type I- and type III- rich tissues. From these results we localized the source of SHG along the single helix of both myosin and collagen. PMID- 19547598 TI - Performance of infrared systems in swimmer detection for maritime security. AB - The detection of swimmer activity in harbor areas around piers and ships is an important aspect of Anti-Terrorism/Force Protection (AT/FP) sensing efforts in the U.S. Navy and Coast Guard. A series of data collections and perception experiments were conducted to validate the use of thermal target acquisition models against swimmer targets. The results were analyzed to derive the discrimination criteria necessary for sensor design for maritime force protection. PMID- 19547599 TI - Automatic channel unmixing for high-throughput quantitative analysis of fluorescence images. AB - Laser-scanning microscopy allows rapid acquisition of multi-channel data, paving the way for high-throughput, high-content analysis of large numbers of images. An inherent problem of using multiple fluorescent dyes is overlapping emission spectra, which results in channel cross-talk and reduces the ability to extract quantitative measurements. Traditional unmixing methods rely on measuring channel cross-talk and using fixed acquisition parameters, but these requirements are not suited to high-throughput processing. Here we present a simple automatic method to correct for channel cross-talk in multi-channel images using image data only. The method is independent of the acquisition parameters but requires some spatial separation between different dyes in the image. We evaluate the method by comparing the cross-talk levels it estimates to those measured directly from a standard fluorescent slide. The method is then applied to a high-throughput analysis pipeline that measures nuclear volumes and relative expression of gene products from three-dimensional, multi-channel fluorescence images of whole Drosophila embryos. Analysis of images before unmixing revealed an aberrant spatial correlation between measured nuclear volumes and the gene expression pattern in the shorter wavelength channel. Applying the unmixing algorithm before performing these analyses removed this correlation. PMID- 19547600 TI - 3-D shape measurement by composite pattern projection and hybrid processing. AB - This article presents a projection system with a novel composite pattern for one shot acquisition of 3D surface shape. The pattern is composed of color encoded stripes and cosinoidal intensity fringes, with parallel arrangement. The stripe edges offer absolute height phases with high accuracy, and the cosinoidal fringes provide abundant relative phases involved in the intensity distribution. Wavelet transform is utilized to obtain the relative phase distribution of the fringe pattern, and the absolute height phases measured by triangulation are combined to calibrate the phase data in unwrapping, so as to eliminate the initial and noise errors and to reduce the accumulation and approximation errors. Numerical simulations are performed to prove the new unwrapping algorithms and actual experiments are carried out to show the validity of the proposed technique for accurate 3- D shape measurement. PMID- 19547601 TI - Surface plasmon mode analysis of nanoscale metallic rectangular waveguide. AB - A detailed study of guided modes in a nanoscale metallic rectangular waveguide is presented by using the effective dielectric constant approach. The guided modes, including both traditional waveguide mode and surface plasmon mode, are investigated for the silver rectangular waveguide. The mode evolution in narrow waveguide is also discussed with the emphasis on the dependence of mode dispersion with waveguide height. Finally, the red-shift of the cutoff wavelength of the fundamental mode is observed when the waveguide height decreases, contrary to the behavior of regular metallic waveguide with PEC boundary. The comprehensive analysis can provide some guideline in the design of subwavelength optical devices based on the dispersion characteristics of metallic rectangular bore. PMID- 19547602 TI - Closed loop, DM diversity-based, wavefront correction algorithm for high contrast imaging systems. AB - High contrast imaging from space relies on coronagraphs to limit diffraction and a wavefront control systems to compensate for imperfections in both the telescope optics and the coronagraph. The extreme contrast required (up to 10(-10) for terrestrial planets) puts severe requirements on the wavefront control system, as the achievable contrast is limited by the quality of the wavefront. This paper presents a general closed loop correction algorithm for high contrast imaging coronagraphs by minimizing the energy in a predefined region in the image where terrestrial planets could be found. The estimation part of the algorithm reconstructs the complex field in the image plane using phase diversity caused by the deformable mirror. This method has been shown to achieve faster and better correction than classical speckle nulling. PMID- 19547603 TI - Amplification and front facet reflectivity of broad area lasers. AB - The effect of front-facet reflectivity on the amplification performance of Broad Area Laser (BAL) diodes in a double-pass configurationis studied experimentally. A method to measure the front facet reflectivities of laser diodes is generalized to BALs. The method is based on fitting a model, with front facet reflectivity as a parameter, to the threshold current vs. external feedback of the diode. Reflectivities of three BAL diodes are measured, and their amplification abilities have been assessed. The tested diodes had amplification factors of 0, 1, and 10 and front facet reflectivities of 12.7 +/-1%, 4.6 +/-0.4%, and 1.2 +/ 0.2% respectively. It is concluded that a front facet modal reflectivity of less than 4.6% is necessary for a BAL to function as an amplifier. PMID- 19547604 TI - Fiber-bragg-grating-based dispersion-compensated and gain-flattened raman fiber Amplifier. AB - In this paper, we propose a novel signal/pump double-pass Raman fiber amplifier using fiber Brag gratings (FBGs). In order to compensate the dispersion slop mismatch among channels in lightwave system, FBGs embedded in different positions along dispersion compensated fiber are used to control the travel length of each WDM signal. Gain equalization can be achieved by optimizing the reflectivity of each FBG. Maximum output power variation among channels is less than +/-0.5 dB after appropriate optimization. Finally, a wavelength division multiplexing (WDM) system using 40-Gb/s x 8 ch non return-to-zero (NRZ) signal transmission in a 100 km transmission fiber is simulated to confirm the system performance. Using proposed dispersion compensation method, it may lead to 2 dB improvement in Q value. Such kind of RFA may find vast applications in WDM system where dispersion management is a crucial issue. PMID- 19547605 TI - Highly efficient all-fiber tunable polarization filter using torsional acoustic wave. AB - We demonstrate an all-fiber tunable polarization filter with high coupling efficiency based on acousto-optic coupling between two optical polarization modes of the LP(01) mode propagating in a highly birefringent single mode optical fiber. An over-coupling between the two polarization modes is realized over the wavelength range from 1530 nm to 1610 nm using traveling torsional acoustic wave. The measured 3-dB optical bandwidth of the filter was 4.8 nm at the wavelength around 1550 nm. The details of the filter transmission and the coupling characteristics are discussed. PMID- 19547606 TI - Optical bistability in subwavelength metallic grating coated by nonlinear material. AB - A developed two-dimensional Finite Difference Time Domain (FDTD) method has been performed to investigate the optical bistability in a subwavelength metallic grating coated by nonlinear material. Different bistability loops have been shown to depend on parameters of the structure. The influences of two key parameters, thickness of nonlinear material and slit width of metallic grating, have been studied in detail. The effect of optical bistability in the structure is explained by Surface Plasmons (SPs) mode and resonant waveguide theory. PMID- 19547607 TI - H(2) impact on Bragg gratings written in N-doped silica-core fiber. AB - The evolution of transmission spectra of Bragg gratings written in an N-doped silica-core fiber in the course of H(2) loading at a pressure of 6 MPa is investigated. It is shown, that penetration of hydrogen molecules in the region of fiber core with written gratings causes irreversible spectrum changes, which do not disappear after subsequent H(2) outcome from the fiber. Bragg gratings' spectra monitoring in the process of H(2) loading is viewed from the angle of photosensitivity mechanisms responsible for formation in N-doped silica-core fibers photoinduced Bragg gratings, capable to operate at very high temperatures. PMID- 19547608 TI - Low-threshold bistability of slow light in photonic-crystal waveguides. AB - We analyze the resonant transmission of light through a photonic-crystal waveguide side coupled to a Kerr nonlinear cavity, and demonstrate how to design the structure geometry for achieving bistability and all-optical switching at ultralow powers in the slow-light regime. We show that the resonance quality factor in such structures scales inversely proportional to the group velocity of light at the resonant frequency and thus grows indefinitely in the slow-light regime. Accordingly, the power threshold required for all-optical switching in such structures scales as a square of the group velocity, rapidly vanishing in the slow-light regime. PMID- 19547609 TI - A conceptual experiment on single-beam coincidence detection with pseudothermal light. AB - Ghost imaging produced by pseudothermal light is commonly obtained by correlating the intensities of two separate beams, neither of which conveys information about the shape of the object to be imaged. The single-beam experiment discussed here, while not exploitable for the practical purpose of reconstructing the shape of a real mask, uses the same mathematical machinery as two-beam experiments; it also suggests that image retrieval by classical light ghost imaging is only a product of normal signal processing and does not involve any "ghost". In addition, the singlebeam setup allows simpler calibration procedures in systematic investigations of the efficiency of coincidence imaging. PMID- 19547610 TI - Analysis of the effects of spherical aberration on ultrafast laser-induced refractive index variation in glass. AB - We propose a comprehensive analysis of the effects that spherical aberration may have on the process of ultrafast laser photowriting in bulk transparent materials and discuss the consequences for the generated refractive index changes. Practical aspects for a longitudinal photowriting configuration are emphasized. Laser-induced index variation in BK7 optical glass and fused silica (a-SiO(2)) affected by spherical aberration are characterized experimentally using phase contrast optical microscopy. Experimental data are matched by analytical equations describing light propagation through dielectric interfaces. Corrective solutions are proposed with a particular focus on the spatial resolution achievable and on the conditions to obtain homogeneously photo-induced waveguides in a longitudinal writing configuration. PMID- 19547611 TI - Soliton percolation in random optical lattices. AB - We introduce soliton percolation phenomena in the nonlinear transport of light packets in suitable optical lattices with random properties. Specifically, we address lattices with a gradient of the refractive index in the transverse plane, featuring stochastic phase or amplitude fluctuations, and we discover the existence of a disorder-induced transition between soliton insulator and soliton conductor regimes. The soliton current is found to reach its maximal value at intermediate disorder levels and to drastically decrease in both, almost regular and strongly disordered lattices. PMID- 19547612 TI - Terahertz photonic crystal resonators in double-metal waveguides. AB - We present the design and the fabrication of photonic crystals with a complete bandgap for TM-modes used as a resonator for terahertz quantum-cascade lasers (QCL), which are lasing around 2.7 THz. The emission of the devices with and without a photonic crystal shows a shift in the emission from the gain maximum to the bandgap of the crystal. The devices are built up by a core, which provides the optical gain, and by a surrounding photonic crystal, which acts as a frequency selective mirror. The whole device is processed into a double-metal waveguide. PMID- 19547613 TI - New photopolymer holographic recording material with sustainable design. AB - Photopolymers that absorb in the visible spectrum are useful for different applications such as in the development of holographic memories, holographic optical elements or as holographic recording media. Photopolymers have an undesirable feature, the toxicity of their components and their low environmental compatibility, particularly if we analyse the life cycle of the devices made with these materials and their interaction with the environment. In this work we developed a new photopolymer with photochemical and holographic features similar to those of the standard material but with an improved design from the environmental point of view. PMID- 19547614 TI - Integrated polarization rotator made of periodic asymmetric buried Ta2O5 / silica sol-gel waveguides. AB - A ridge waveguide technology exhibiting high polarization dependency is developed for new efficient multi-section passive polarization rotator applications. In the presented configuration, the calculated mode coupling between the waveguide sections is very efficient and allows a polarization rotation with a high extinction ratio at lambda=1,55 mum. Experimental results show efficient polarization rotation with low cross-talk levels (-16dB) and no significant excess losses between sections. However, the overall transmission efficiency is limited by propagation losses and coupling losses to standard optical fibers. PMID- 19547615 TI - Ultimate vertical Fabry-Perot cavity based on single-layer photonic crystal mirrors. AB - Vertical Fabry Perot cavities (VFPC) have been extensively studied, especially for the realization of vertical-cavity surface emitting lasers (VCSELs). They are traditionally composed of two Distributed Bragg Reflectors (DBR) which reflectivity has to be sufficient in order to obtain highly resonant cavity, which is particularly necessary for laser emission in VCSELs. As a consequence, DBRs consist generally in very thick layer stacks. In this paper, we demonstrate the smallest conceivable high Q vertical Fabry-Perot cavity, using ultra-thin and highly-efficient photonic crystal slab mirrors instead of conventional DBRs, which enable moreover a control of the polarization. PMID- 19547616 TI - Multi-wavelength-switchable and Uniform Erbium-doped Fiber Laser Using Unbalanced In-line Sagnac Interferometer. AB - We have proposed and demonstrated a multi-wavelength-switchable and uniform erbium-doped fiber laser using unbalanced in-line Sagnac Interferometer. By employing this simple scheme and through the proper control of the polarization controller, we were able to achieve uniform multi-wavelength operation of up to 84 laser lines with the signal-to- noise ratio over 20dB and 0.8-nm wavelength switching at room temperature. Again, we generated more than 300 lines, 0.1-m wavelength switching and good power stability (/= 95%. RESULTS: The study cohort included 40 adult patients. 18,562 rSO(2) values were collected during OLV. The rSO(2) was >/= baseline at 3,593 of the 18,562 data points (19%). The rSO2 was 0-9 /= 25% of the duration of OLV. These patients were older (63.7 +/- 10.2 vs 54.6 +/- 9.8 years, P<0.025), weighed more (95.8 +/- 17.4 vs 82.6 +/- 14.6 kgs, P=0.038), and were more likely to be ASA III vs II (7 of 8 versus 25 of 32, relative risk 1.75) than the remainder of the cohort. CONCLUSIONS: Significant changes in rSO2 occur during OLV for thoracic surgical procedures. Future studies are needed to determine the impact of such changes on the postoperative course of these patients. PMID- 19547694 TI - Recurrent genomic gains in preinvasive lesions as a biomarker of risk for lung cancer. AB - Lung carcinoma development is accompanied by field changes that may have diagnostic significance. We have previously shown the importance of chromosomal aneusomy in lung cancer progression. Here, we tested whether genomic gains in six specific loci, TP63 on 3q28, EGFR on 7p12, MYC on 8q24, 5p15.2, and centromeric regions for chromosomes 3 (CEP3) and 6 (CEP6), may provide further value in the prediction of lung cancer. Bronchial biopsy specimens were obtained by LIFE bronchoscopy from 70 subjects (27 with prevalent lung cancers and 43 individuals without lung cancer). Twenty six biopsies were read as moderate dysplasia, 21 as severe dysplasia and 23 as carcinoma in situ (CIS). Four-micron paraffin sections were submitted to a 4-target FISH assay (LAVysion, Abbott Molecular) and reprobed for TP63 and CEP 3 sequences. Spot counts were obtained in 30-50 nuclei per specimen for each probe. Increased gene copy number in 4 of the 6 probes was associated with increased risk of being diagnosed with lung cancer both in unadjusted analyses (odds ratio = 11, p<0.05) and adjusted for histology grade (odds ratio = 17, p<0.05). The most informative 4 probes were TP63, MYC, CEP3 and CEP6. The combination of these 4 probes offered a sensitivity of 82% for lung cancer and a specificity of 58%. These results indicate that specific cytogenetic alterations present in preinvasive lung lesions are closely associated with the diagnosis of lung cancer and may therefore have value in assessing lung cancer risk. PMID- 19547696 TI - [Not Available]. AB - Spigelian hernia occurs through slit like defect in the anterior abdominal wall adjacent to the semilunar line. Most of spigelian hernias occur in the lower abdomen where the posterior sheath is deficient. The hernia ring is a well defined defect in the transverses aponeurosis. The hernial sac, surrounded by extraperitoneal fatty tissue, is often interparietal passing through the transversus and the internal oblique aponeuroses and then spreading out beneath the intact aponeurosis of the external oblique. Spigelian hernia is in itself very rare and more over it is difficult to diagnose clinically. It has been estimated that it constitutes 0.12% of abdominal wall hernias. The spigelian hernia has been repaired by both conventional and laparoscopic approach. Laparoscopic management of spigelian hernia is well established. Most of the authors have managed it by transperitoneal approach either by placing the mesh in intraperitoneal position or by raising the peritoneal flap and placing the mesh in extraperitoneal space. There have also been case reports of management of spigelian hernia by total extraperitoneal approach. We retrospectively reviewed our experience of ten patients between 1997 and 2007. Eight patients (8/10) presented with abdominal pain and two patients (2/10) were asymptomatic. In six patients (6/10) we performed an intraperitoneal onlay IPOM repair, in two patients (2/10) transabdominal preperitoneal repair (TAPP), and in two (2/10) total extraperitoneal repair (TEP). There were no recurrences, or other morbidity at mean follow up period of 3.2 years (range 6 months to 10 years). PMID- 19547697 TI - [Not Available]. AB - Bronchobiliary fistula is a very rare complication of liver abscess. It presents with biliptysis (bile in cough), and chronic cough. Here we present a case of intractable biliptysis from a bronchobiliary fistula secondary to a liver abscess with biliary obstruction. PMID- 19547698 TI - [Not Available]. AB - BACKGROUND: Laparoscopic fundoplication (LF) has become the operation of choice for patients who need surgery for gastro esophageal reflux disease (GERD). Several studies have shown that the long-term results with surgery for GERD are better than medical therapy. In this retrospective study, we outline our experience with LF over an 8 year period. We analyzed factors that would affect the results of surgery and help in a better selection of patients for the operation. MATERIALS AND METHODS: From 1999 to 2007, 107 patients underwent a LF. Eighty five patients had surgery for GERD and form the basis of this article. The other 22 patients had paraesophageal hernias and were excluded from the study. Pre-operative evaluation consisted of endoscopy, a barium study, esophageal manometry and 24h pH monitoring. Patients were followed up every 3rd month for the 1st year, twice in the 2nd year and then annually. Follow up was by personal interview or telephonic conversation. At the last follow up the results of surgery were graded as good or poor as per a scoring system. Those with a poor result were evaluated and re-operation advised when an anatomical problem caused the poor result. Subjective, objective and technical variables were analyzed which could affect the outcome of surgery. RESULTS: In 84 patients, the operation was completed by laparoscopic access. One patient with bleeding was converted to open surgery. There were 5 intra-operative complications; 3 pnemothoracis, 1 esophageal perforation and 1 gastric fundus perforation. There was no mortality. Two patients underwent re-operation, 1 for delayed gastric emptying and 1 for dysphagia. Seventy four patients have been followed up from 7 months to 8 years. Eleven have been lost to follow up. Fifty seven patients (77%) have had a good result from surgery. Seventeen (23%) had a poor result; of these there were 4 wrap failures, 1 delayed gastric emptying and 1 excessive gas bloat as the cause. In 11 patients, there was no apparent cause of a poor result. Individual variables which predicted a good response to surgery (P<0.5); were a good response to proton pump inhibitors (PPis), volume reflux and a pH score of more than 14. CONCLUSION: LF gives good long-term relief of symptoms in patients with GERD. Strict selection criteria are necessary to optimize the results of surgery. Poor selection will result in a patient who is no better, or often worse than before surgery. PMID- 19547699 TI - Disassembly of shank and homer synaptic clusters is driven by soluble beta amyloid(1-40) through divergent NMDAR-dependent signalling pathways. AB - Disruption of the postsynaptic density (PSD), a network of scaffold proteins located in dendritic spines, is thought to be responsible for synaptic dysfunction and loss in early-stage Alzheimer's disease (AD). Extending our previous demonstration that derangement of the PSD by soluble amyloid-beta (Abeta) involves proteasomal degradation of PSD-95, a protein important for ionotropic glutamate receptor trafficking, we now show that Abeta also disrupts two other scaffold proteins, Homer1b and Shank1, that couple PSD-95 with ionotropic and metabotropic glutamate receptors. Treatment of fronto-cortical neurons with soluble Abeta results in rapid (within 1 h) and significant thinning of the PSD, decreased synaptic levels of Homer1b and Shank1, and reduced synaptic mGluR1 levels. We show that de novo protein synthesis is required for the declustering effects of Abeta on Homer1b (but not Shank1) and that, in contrast to PSD-95, Abeta-induced Homer1b and Shank1 cluster disassembly does not depend on proteasome activity. The regulation of Homer1b and Shank1 by Abeta diverges in two other respects: i) whereas the activity of both NMDAR and VDCC is required for Abeta-induced declustering of Homer1b, Abeta-induced declustering of Shank1 only requires NMDAR activity; and ii) whereas the effects of Abeta on Homer1b involve engagement of the PI-3K pathway and calcineurin phosphatase (PP2B) activity, those on Shank1 involve activation of the ERK pathway. In summary, soluble Abeta recruits discrete signalling pathways to rapidly reduce the synaptic localization of major components of the PSD and to regulate the availability of mGluR1 in the synapse. PMID- 19547700 TI - Cell therapy attenuates cardiac dysfunction post myocardial infarction: effect of timing, routes of injection and a fibrin scaffold. AB - BACKGROUND: Cell therapy approaches for biologic cardiac repair hold great promises, although basic fundamental issues remain poorly understood. In the present study we examined the effects of timing and routes of administration of bone marrow cells (BMC) post-myocardial infarction (MI) and the efficacy of an injectable biopolymer scaffold to improve cardiac cell retention and function. METHODOLOGY/PRINCIPAL FINDINGS: (99m)Tc-labeled BMC (6 x 10(6) cells) were injected by 4 different routes in adult rats: intravenous (IV), left ventricular cavity (LV), left ventricular cavity with temporal aorta occlusion (LV(+)) to mimic coronary injection, and intramyocardial (IM). The injections were performed 1, 2, 3, or 7 days post-MI and cell retention was estimated by gamma-emission counting of the organs excised 24 hs after cell injection. IM injection improved cell retention and attenuated cardiac dysfunction, whereas IV, LV or LV* routes were somewhat inefficient (<1%). Cardiac BMC retention was not influenced by timing except for the IM injection that showed greater cell retention at 7 (16%) vs. 1, 2 or 3 (average of 7%) days post-MI. Cardiac cell retention was further improved by an injectable fibrin scaffold at day 3 post-MI (17 vs. 7%), even though morphometric and function parameters evaluated 4 weeks later displayed similar improvements. CONCLUSIONS/SIGNIFICANCE: These results show that cells injected post-MI display comparable tissue distribution profile regardless of the route of injection and that there is no time effect for cardiac cell accumulation for injections performed 1 to 3 days post-MI. As expected the IM injection is the most efficient for cardiac cell retention, it can be further improved by co injection with a fibrin scaffold and it significantly attenuates cardiac dysfunction evaluated 4 weeks post myocardial infarction. These pharmacokinetic data obtained under similar experimental conditions are essential for further development of these novel approaches. PMID- 19547701 TI - Chlamydia pneumoniae hides inside apoptotic neutrophils to silently infect and propagate in macrophages. AB - BACKGROUND: Intracellular pathogens have developed elaborate strategies for silent infection of preferred host cells. Chlamydia pneumoniae is a common pathogen in acute infections of the respiratory tract (e.g. pneumonia) and associated with chronic lung sequelae in adults and children. Within the lung, alveolar macrophages and polymorph nuclear neutrophils (PMN) are the first line of defense against bacteria, but also preferred host phagocytes of chlamydiae. METHODOLOGY/PRINCIPAL FINDINGS: We could show that C. pneumoniae easily infect and hide inside neutrophil granulocytes until these cells become apoptotic and are subsequently taken up by macrophages. C. pneumoniae infection of macrophages via apoptotic PMN results in enhanced replicative activity of chlamydiae when compared to direct infection of macrophages, which results in persistence of the pathogen. Inhibition of the apoptotic recognition of C. pneumoniae infected PMN using PS- masking Annexin A5 significantly lowered the transmission of chlamydial infection to macrophages. Transfer of apoptotic C. pneumoniae infected PMN to macrophages resulted in an increased TGF-ss production, whereas direct infection of macrophages with chlamydiae was characterized by an enhanced TNF-alpha response. CONCLUSIONS/SIGNIFICANCE: Taken together, our data suggest that C. pneumoniae uses neutrophil granulocytes to be silently taken up by long-lived macrophages, which allows for efficient propagation and immune protection within the human host. PMID- 19547702 TI - [Not Available]. AB - Malaria's global impact is expansive and includes the extremes of the healthcare system ranging from international travelers returning to nonendemic regions with tertiary referral medical care to residents in hyperendemic regions without access to medical care. Implementation of prompt and accurate diagnosis is needed to curb the expanding global impact of malaria associated with ever-increasing antimalarial drug resistance. Traditionally, malaria is diagnosed using clinical criteria and/or light microscopy even though both strategies are clearly inadequate in many healthcare settings. Hand held immunochromatographic rapid diagnostic tests (RDTs) have been recognized as an ideal alternative method for diagnosing malaria. Numerous malaria RDTs have been developed and are widely available; however, an assortment of issues related to these products have become apparent. This review provides a summary of RDT including effectiveness and strategies to select the ideal RDT in varying healthcare settings. PMID- 19547703 TI - [Not Available]. AB - Chagas disease which is caused by the parasite Trypanosoma cruzi is an important cause of cardiomyopathy in Latin America. In later stages chagasic cardiomyopathy is associated with congestive heart failure which is often refractory to medical therapy. In these individuals heart transplantation has been attempted. However, this procedure is fraught with many problems attributable to the surgery and the postsurgical administration of immunosuppressive drugs. Studies in mice suggest that the transplantation of bone-marrow-derived cells ameliorates the inflammation and fibrosis in the heart associated with this infection. Cardiac magnetic resonance imaging reveals that bone marrow transplantation ameliorates the infection induced right ventricular enlargement. On the basis of these animal studies the safety of autologous bone marrow transplantation has been assessed in patients with chagasic end-stage heart disease. The initial results are encouraging and more studies need to be performed. PMID- 19547704 TI - Blind deblurring reconstruction technique with applications in PET imaging. AB - We developed an empirical PET model taking into account system blurring and a blind iterative reconstruction scheme that estimates both the actual image and the point spread function of the system. Reconstruction images of high quality can be acquired by using the proposed reconstruction technique for both synthetic and experimental data. In the synthetic data study, the algorithm reduces image blurring and preserves the edges without introducing extra artifacts. The localized measurement shows that the performance of the reconstruction image improved by up to 100%. In experimental data studies, the contrast and quality of reconstruction is substantially improved. The proposed method shows promise in tumor localization and quantification. PMID- 19547705 TI - [Not Available]. AB - In our recent work, Monte Carlo Cross Validation Stacked Regression (MCCVSR) is proposed to achieve automatic optimization of spectral interval selection in multivariate calibration. Though MCCVSR performs well in normal conditions, it is still necessary to improve it for more general applications. According to the well-known principle of "garbage in, garbage out (GIGO)", as a precise ensemble method, MCCVSR might be influenced by outlying and very bad submodels. In this paper, a statistical test is designed to exclude the ruinous submodels from the ensemble learning process, therefore, the combination process becomes more reliable. Though completely automated, the proposed method is adjustable according to the nature of the data analyzed, including the size of training samples, resolution of spectra and quantitative potentials of the submodels. The effectiveness of the submodel refining is demonstrated by the investigation of a real standard data. PMID- 19547706 TI - Segmentation of myocardial boundaries in tagged cardiac MRI using active contours: a gradient-based approach integrating texture analysis. AB - The noninvasive assessment of cardiac function is of first importance for the diagnosis of cardiovascular diseases. Among all medical scanners only a few enables radiologists to evaluate the local cardiac motion. Tagged cardiac MRI is one of them. This protocol generates on Short-Axis (SA) sequences a dark grid which is deformed in accordance with the cardiac motion. Tracking the grid allows specialists a local estimation of cardiac geometrical parameters within myocardium. The work described in this paper aims to automate the myocardial contours detection in order to optimize the detection and the tracking of the grid of tags within myocardium. The method we have developed for endocardial and epicardial contours detection is based on the use of texture analysis and active contours models. Texture analysis allows us to define energy maps more efficient than those usually used in active contours methods where attractor is often based on gradient and which were useless in our case of study, for quality of tagged cardiac MRI is very poor. PMID- 19547707 TI - Molecular determinants of juvenile hormone action as revealed by 3D QSAR analysis in Drosophila. AB - BACKGROUND: Postembryonic development, including metamorphosis, of many animals is under control of hormones. In Drosophila and other insects these developmental transitions are regulated by the coordinate action of two principal hormones, the steroid ecdysone and the sesquiterpenoid juvenile hormone (JH). While the mode of ecdysone action is relatively well understood, the molecular mode of JH action remains elusive. METHODOLOGY/PRINCIPAL FINDINGS: To gain more insights into the molecular mechanism of JH action, we have tested the biological activity of 86 structurally diverse JH agonists in Drosophila melanogaster. The results were evaluated using 3D QSAR analyses involving CoMFA and CoMSIA procedures. Using this approach we have generated both computer-aided and species-specific pharmacophore fingerprints of JH and its agonists, which revealed that the most active compounds must possess an electronegative atom (oxygen or nitrogen) at both ends of the molecule. When either of these electronegative atoms are replaced by carbon or the distance between them is shorter than 11.5 A or longer than 13.5 A, their biological activity is dramatically decreased. The presence of an electron-deficient moiety in the middle of the JH agonist is also essential for high activity. CONCLUSIONS/SIGNIFICANCE: The information from 3D QSAR provides guidelines and mechanistic scope for identification of steric and electrostatic properties as well as donor and acceptor hydrogen-bonding that are important features of the ligand-binding cavity of a JH target protein. In order to refine the pharmacophore analysis and evaluate the outcomes of the CoMFA and CoMSIA study we used pseudoreceptor modeling software PrGen to generate a putative binding site surrogate that is composed of eight amino acid residues corresponding to the defined molecular interactions. PMID- 19547708 TI - Histamine H(3) receptor-mediated signaling protects mice from cerebral malaria. AB - BACKGROUND: Histamine is a biogenic amine that has been shown to contribute to several pathological conditions, such as allergic conditions, experimental encephalomyelitis, and malaria. In humans, as well as in murine models of malaria, increased plasma levels of histamine are associated with severity of infection. We reported recently that histamine plays a critical role in the pathogenesis of experimental cerebral malaria (CM) in mice infected with Plasmodium berghei ANKA. Histamine exerts its biological effects through four different receptors designated H1R, H2R, H3R, and H4R. PRINCIPAL FINDINGS: In the present work, we explored the role of histamine signaling via the histamine H3 receptor (H3R) in the pathogenesis of murine CM. We observed that the lack of H3R expression (H3R(-/-) mice) accelerates the onset of CM and this was correlated with enhanced brain pathology and earlier and more pronounced loss of blood brain barrier integrity than in wild type mice. Additionally tele-methylhistamine, the major histamine metabolite in the brain, that was initially present at a higher level in the brain of H3R(-/-) mice was depleted more quickly post-infection in H3R(-/-) mice as compared to wild-type counterparts. CONCLUSIONS: Our data suggest that histamine regulation through the H3R in the brain suppresses the development of CM. Thus modulating histamine signaling in the central nervous system, in combination with standard therapies, may represent a novel strategy to reduce the risk of progression to cerebral malaria. PMID- 19547709 TI - Alpha-helical protein networks are self-protective and flaw-tolerant. AB - Alpha-helix based protein networks as they appear in intermediate filaments in the cell's cytoskeleton and the nuclear membrane robustly withstand large deformation of up to several hundred percent strain, despite the presence of structural imperfections or flaws. This performance is not achieved by most synthetic materials, which typically fail at much smaller deformation and show a great sensitivity to the existence of structural flaws. Here we report a series of molecular dynamics simulations with a simple coarse-grained multi-scale model of alpha-helical protein domains, explaining the structural and mechanistic basis for this observed behavior. We find that the characteristic properties of alpha helix based protein networks are due to the particular nanomechanical properties of their protein constituents, enabling the formation of large dissipative yield regions around structural flaws, effectively protecting the protein network against catastrophic failure. We show that the key for these self protecting properties is a geometric transformation of the crack shape that significantly reduces the stress concentration at corners. Specifically, our analysis demonstrates that the failure strain of alpha-helix based protein networks is insensitive to the presence of structural flaws in the protein network, only marginally affecting their overall strength. Our findings may help to explain the ability of cells to undergo large deformation without catastrophic failure while providing significant mechanical resistance. PMID- 19547710 TI - Expression of Pseudomonas aeruginosa CupD fimbrial genes is antagonistically controlled by RcsB and the EAL-containing PvrR response regulators. AB - Pseudomonas aeruginosa is a gram-negative pathogenic bacterium with a high adaptive potential that allows proliferation in a broad range of hosts or niches. It is also the causative agent of both acute and chronic biofilm-related infections in humans. Three cup gene clusters (cupA-C), involved in the assembly of cell surface fimbriae, have been shown to be involved in biofilm formation by the P. aeruginosa strains PAO1 or PAK. In PA14 isolates, a fourth cluster, named cupD, was identified within a pathogenicity island, PAPI-I, and may contribute to the higher virulence of this strain. Expression of the cupA genes is controlled by the HNS-like protein MvaT, whereas the cupB and cupC genes are under the control of the RocS1A1R two-component system. In this study, we show that cupD gene expression is positively controlled by the response regulator RcsB. As a consequence, CupD fimbriae are assembled on the cell surface, which results in a number of phenotypes such as a small colony morphotype, increased biofilm formation and decreased motility. These behaviors are compatible with the sessile bacterial lifestyle. The balance between planktonic and sessile lifestyles is known to be linked to the intracellular levels of c-di-GMP with high levels favoring biofilm formation. We showed that the EAL domain-containing PvrR response regulator counteracts the activity of RcsB on cupD gene expression. The action of PvrR is likely to involve c-di-GMP degradation through phosphodiesterase activity, confirming the key role of this second messenger in the balance between bacterial lifestyles. The regulatory network between RcsB and PvrR remains to be elucidated, but it stands as a potential model system to study how the equilibrium between the two lifestyles could be influenced by therapeutic agents that favor the planktonic lifestyle. This would render the pathogen accessible for the immune system or conventional antibiotic treatment. PMID- 19547711 TI - Reliability at the lower limits of HIV-1 RNA quantification in clinical samples: a comparison of RT-PCR versus bDNA assays. AB - INTRODUCTION: To explore whether an assay change was responsible for an increasing proportion of patients with undetectable HIV viral loads at our urban HIV clinic, we selected highly stable patients, examining their viral loads before and after changing assays. We compared the proportion with detectable viremia during RT-PCR vs. bDNA periods. METHODOLOGY/PRINCIPAL FINDINGS: We selected patients with > or =1 viral loads assessed during both RT-PCR and bDNA periods. We included patients with stable CD4 counts, excluding patients with viral loads > or =1,000 copies/ml or any significant changes in therapy. Out of 4500 clinic patients, 419 patients (1588 viral loads) were included. 39% of viral loads were reported as detectable by RT-PCR vs. 5% reported as detectable by bDNA. The mean coefficient of variation was higher before vs. after assay change. We found an odds' ratio of 16.7 for having a viral load >75 copies/ml during the RT-PCR vs. bDNA periods. DISCUSSION: These data support previous reports, suggesting that bDNA may more reliably discriminate between viral suppression and low level viremia in stable patients on therapy. Low-level viremia, noted more with RT-PCR, may promote unneeded testing, while differences in viral load reliability may impact antiretroviral trial and quality assurance endpoints. Commonly used plasma separator tubes may differentially affect RT-PCR and bDNA results. PMID- 19547712 TI - Understanding physiological and degenerative natural vision mechanisms to define contrast and contour operators. AB - BACKGROUND: Dynamical systems like neural networks based on lateral inhibition have a large field of applications in image processing, robotics and morphogenesis modeling. In this paper, we will propose some examples of dynamical flows used in image contrasting and contouring. METHODOLOGY: First we present the physiological basis of the retina function by showing the role of the lateral inhibition in the optical illusions and pathologic processes generation. Then, based on these biological considerations about the real vision mechanisms, we study an enhancement method for contrasting medical images, using either a discrete neural network approach, or its continuous version, i.e. a non-isotropic diffusion reaction partial differential system. Following this, we introduce other continuous operators based on similar biomimetic approaches: a chemotactic contrasting method, a viability contouring algorithm and an attentional focus operator. Then, we introduce the new notion of mixed potential Hamiltonian flows; we compare it with the watershed method and we use it for contouring. CONCLUSIONS: We conclude by showing the utility of these biomimetic methods with some examples of application in medical imaging and computed assisted surgery. PMID- 19547713 TI - Ferromagnetic levan composite: an affinity matrix to purify lectin. AB - A simple and inexpensive procedure used magnetite and levan to synthesize a composite recovered by a magnetic field. Lectins from Canavalia ensiformis (Con A) and Cratylia mollis (Cramoll 1 and Cramoll 1, 4) did bind specifically to composite. The magnetic property of derivative favored washing out contaminating proteins and recovery of pure lectins with glucose elution. Cramoll 1 was purified by this affinity binding procedure in two steps instead of a previous three-step protocol with ammonium sulfate fractionation, affinity chromatography on Sephadex G-75, and ion exchange chromatography through a CM-cellulose column. PMID- 19547714 TI - [Not Available]. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is currently being investigated as a therapeutic agent for a variety of malignancies, as it triggers apoptosis specifically in transformed cells. However, TRAIL use as a stand alone therapeutic is hampered by the fact that many primary tumor cells are resistant to TRAIL-mediated apoptosis. Here, we investigated the extent to which pretreatment of TRAIL-resistant primary B-cell chronic lymphocytic leukemia (B CLL) cells with histone deacetylase inhibitors (HDACis) could render them susceptible to killing by TRAIL. We found that HDAC inhibition in B-CLL cells led to increased TRAIL receptor expression, increased caspase activation, decreased expression of antiapoptotic regulators such as Bcl-2, and ultimately, enhanced TRAIL-induced apoptosis. Importantly, untransformed peripheral blood mononuclear cells remained largely resistant to TRAIL, even in the presence of HDACis. These results suggest that combination therapies using HDAC inhibition and TRAIL could prove beneficial for the treatment of B-CLL. PMID- 19547715 TI - [Not Available]. AB - Tumor cells that are nonsensitive to anticancer drugs frequently have a multidrug resistant (MDR) phenotype. Many studies with cell lines and patient material have been done to investigate the impact of different resistance markers at protein and mRNA level in drug resistance but with contradictory outcome. In the present study, 26 well-characterised patient-derived non-small cell lung cancer xenografts were used. The known chemosensitivity to etoposide, carboplatin, gemcitabine, paclitaxel and erlotinib was compared to the protein and mRNA expression of BCRP, LRP, MDR1, and MRP1. Further, four of these xenografts were short-term treated to analyse possible regulation mechanisms after therapeutic interventions. We found a borderline correlation between the bcrp mRNA expression and the response of xenografts to etoposide. All other constitutive mRNA and protein expression levels were not correlated to any drug response and were not significantly influenced by a short term treatment. The present results indicate that the expression levels of MDR proteins and mRNA investigated do not play an important role in the chemoresistance of NSCLC in the in vivo situation. PMID- 19547716 TI - [Not Available]. AB - There is growing evidence to suggest that chagasic myocardia are exposed to sustained oxidative stress induced injuries that may contribute to disease progression. Trypanosoma cruzi invasion- and replication-mediated cellular injuries and immune-mediated cytotoxic reactions are the common source of reactive oxygen species (ROS) during acute infection. Mitochondria are proposed to be the major source of ROS in chronic chagasic hearts. However, it has not been established yet, whether mitochondrial dysfunction is a causative factor in chagasic cardiomyopathy or a consequence of other pathological events. A better understanding of oxidative stress in relation to cardiac tissue damage would be useful in the evaluation of its true role in the pathogenesis of Chagas disease and other heart diseases. In this review, we discuss the evidence for increased oxidative stress in chagasic disease, with emphasis on mitochondrial abnormalities, and its role in sustaining oxidative stress in myocardium. PMID- 19547719 TI - Molecular effects on boundary condition in micronanoliquid flows. AB - We experimentally investigated molecular effects of the slipno-slip boundary condition of Newtonian liquids in micro- and nanochannels as small as 350 nm. The slip was measurable for channels smaller than approximately 2 mum. The amount of slip is found to be independent of the channel size, but is a function of the shear rate, the type of liquid (polar or nonpolar molecular structure), and the morphology of the solid surface (molecular-level smoothness). PMID- 19547717 TI - Lornoxicam suppresses recurrent herpetic stromal keratitis through down regulation of nuclear factor-kappaB: an experimental study in mice. AB - PURPOSE: We designed the current study to determine the protective effects of lornoxicam, a cyclooxygenase (COX) inhibitor, on recurrent herpetic stromal keratitis (HSK) and the nuclear factor-kappaB (NF-kappaB)-mediated mechanism in mice. METHODS: A corneal latent herpes simplex virus-1 (HSV-1) infected mouse model was established. Six weeks later, Ultraviolet B (UVB) irradiation induced the recurrence. Corneal swabs were obtained and cultured with indicator cells to determine shedding of the virus. Lornoxicam was administered intraperitoneally daily, beginning one day before irradiation and lasting for seven days. Saline treated and mock-infected control groups were also studied at the same time. Development of corneal inflammation and opacity was scored. Immunohistochemical staining and an electrophoretic mobility shift assay were performed to evaluate the effect of lornoxicam on NF-kappaB activation in the corneal tissues. The levels of tumor necrosis factor-alpha (TNF-alpha) in the cornea were determined by an enzyme-linked immunosorbent assay (ELISA). RESULTS: HSV-1 reactivation induced stromal edema and opacification concomitantly with elevated activation of NF-kappaB and elevated production of TNF-alpha. Lornoxicam treatment significantly decreased the incidence of recurrent HSK, attenuated the corneal opacity scores, and also effectively suppressed both NF-kappaB activation and TNF alpha expression in biological analysis. Histopathology examination revealed a reduced immunostaining positive cell density for NF-kappaB in the cornea from lornoxicam-treated mice as well as a diminished inflammatory response. CONCLUSIONS: Lornoxicam exerts protective effects against HSK, presumably through the down-regulation of NF-kappaB activation. PMID- 19547720 TI - Swimming speeds of filaments in nonlinearly viscoelastic fluids. AB - Many micro-organisms swim through gels and non-Newtonian fluids in their natural environments. In this paper, we focus on micro-organisms which use flagella for propulsion. We address how swimming velocities are affected in nonlinearly viscoelastic fluids by examining the problem of an infinitely long cylinder with arbitrary beating motion in the Oldroyd-B fluid. We solve for the swimming velocity in the limit in which deflections of the cylinder from its straight configuration are small relative to the radius of the cylinder and the wavelength of the deflections; furthermore, the radius of the cylinder is small compared to the wavelength of deflections. We find that swimming velocities are diminished by nonlinear viscoelastic effects. We apply these results to examine what types of swimming motions can produce net translation in a nonlinear fluid, comparing to the Newtonian case, for which Purcell's "scallop" theorem describes how time reversibility constrains which swimming motions are effective. We find that a leading order violation of the scallop theorem occurs for reciprocal motions in which the backward and forward strokes occur at different rates. PMID- 19547718 TI - Presence and regulation of cannabinoid receptors in human retinal pigment epithelial cells. AB - PURPOSE: Cannabinoid receptors have been detected in neuron cells and proposed as potential therapeutic agents in neurodegenerative disorders because of their involvement in controlling neural cell survival and death. However, their presence and role in human retinal pigment epithelial (RPE) cells, which play a key role in initiating and developing age related macular degeneration (ARMD), have never been investigated. Here we analyzed the expression of and changes in cannabinoid receptors (CB1 and CB2) and one enzyme responsible for endocannabinoid hydrolysis, fatty acid amide hydrolase (FAAH), in RPE cell oxidative damage process, a cellular model of ARMD. METHODS: Primary human RPE cells and cells from the ARPE-19 cell line were cultured and exposed to H2O2 for 24 h to induce oxidative damage. Real time RT-PCR, immunofluorescent staining, and western blot methods were performed to study the expression of and changes in CB1 and CB2 receptors, and FAAH. Cell viability and reactive oxygen species (ROS) production were measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and a dichlorofluorescein (DCF) assay, respectively. PI3K/Akt and ERK1/2 protein expression and activation of signaling molecules were assessed by western blot analysis. RESULTS: By using real time RT-PCR, immunofluorescent staining and western blot methods, we showed that human RPE cells express CB1, CB2, and FAAH. Meanwhile, oxidative stress can upregulate CB1 and CB2 receptor expression, and downregulate FAAH expression. The CB1/CB2 receptor agonist, CP55,940, and the CB2 receptor agonist, JWH015 significantly protected RPE cells from oxidative damage. In addition, CP55,940 significantly reduced the levels of intracellular ROS, strengthened oxidative stress-induced activation of PI3K/Akt and reduced activation of the ERK1/2 signal pathway. CONCLUSIONS: The results demonstrate the expression and regulation of CB1 and CB2 receptors and FAAH in human RPE cells. The modulation of cannabinoid receptor tone warrants consideration for future therapeutic strategies of ARMD. PMID- 19547721 TI - Permeability calculations in three-dimensional isotropic and oriented fiber networks. AB - Hydraulic permeabilities of fiber networks are of interest for many applications and have been studied extensively. There is little work, however, on permeability calculations in three-dimensional random networks. Computational power is now sufficient to calculate permeabilities directly by constructing artificial fiber networks and simulating flow through them. Even with today's high-performance computers, however, such an approach would be infeasible for large simulations. It is therefore necessary to develop a correlation based on fiber volume fraction, radius, and orientation, preferably by incorporating previous studies on isotropic or structured networks. In this work, the direct calculations were performed, using the finite element method, on networks with varying degrees of orientation, and combinations of results for flows parallel and perpendicular to a single fiber or an array thereof, using a volume-averaging theory, were compared to the detailed analysis. The detailed model agreed well with existing analytical solutions for square arrays of fibers up to fiber volume fractions of 46% for parallel flow and 33% for transverse flow. Permeability calculations were then performed for isotropic and oriented fiber networks within the fiber volume fraction range of 0.3%-15%. When drag coefficients for spatially periodic arrays were used, the results of the volume-averaging method agreed well with the direct finite element calculations. On the contrary, the use of drag coefficients for isolated fibers overpredicted the permeability for the volume fraction range that was employed. We concluded that a weighted combination of drag coefficients for spatially periodic arrays of fibers could be used as a good approximation for fiber networks, which further implies that the effect of the fiber volume fraction and orientation on the permeability of fiber networks are more important than the effect of local network structure. PMID- 19547723 TI - Flow-structure interaction effects on a jet emanating from a flexible nozzle. AB - In recent years, a wide variety of applications have been found for the use of pulsed jets in the area of flow control. The goal of the current study was to identify the flow field and mixing characteristics associated with an incompressible elongated jet emitted from a flexible nozzle. The shape of the nozzle was that of a high aspect ratio jet deforming from a fully opened to a completely closed configuration. The jet was characterized by a pulsatile flow that was self-excited by the motion of the flexible tube. The frequency of excitation was found to be between 150 and 175 Hz and the Strouhal number (nondimensional frequency) varied from 0.17 to 0.45. The jet flow was dominated by vortices that were shed from the nozzle with an axis parallel to the major axis. The vortices in the near field were quasi-two-dimensional so that measurements performed at the center plane represented the dynamics of the entire vortex. The nozzle excited two different modes depending on the tension applied to the flexible nozzle and the volumetric flow through it. The first was a flapping mode, which was associated with alternate shedding of vortices. This caused strong steering of the jet to one side or the other. The second mode was a symmetric mode that was associated with the formation of counter-rotating vortex pairs. Turbulence and jet spread in the measured planes were much larger in the first mode than the second one. PMID- 19547722 TI - A model of transluminal flow of an anti-HIV microbicide vehicle: Combined elastic squeezing and gravitational sliding. AB - ELASTOHYDRODYNAMIC LUBRICATION OVER SOFT SUBSTRATES IS OF IMPORTANCE IN A NUMBER OF BIOMEDICAL PROBLEMS: From lubrication of the eye surface by the tear film, to lubrication of joints by synovial fluid, to lubrication between the pleural surfaces that protect the lungs and other organs. Such flows are also important for the drug delivery functions of vehicles for anti-HIV topical microbicides. These are intended to inhibit transmission into vulnerable mucosa, e.g., in the vagina. First generation prototype microbicides have gel vehicles, which spread after insertion and coat luminal surfaces. Effectiveness derives from potency of the active ingredients and completeness and durability of coating. Delivery vehicle rheology, luminal biomechanical properties, and the force due to gravity influence the coating mechanics. We develop a framework for understanding the relative importance of boundary squeezing and body forces on the extent and speed of the coating that results. A single dimensionless number, independent of viscosity, characterizes the relative influences of squeezing and gravitational acceleration on the shape of spreading in the Newtonian case. A second scale, involving viscosity, determines the spreading rate. In the case of a shear thinning fluid, the Carreau number also plays a role. Numerical solutions were developed for a range of the dimensionless parameter and compared well with asymptotic theory in the limited case where such results can be obtained. Results were interpreted with respect to trade-offs between wall elasticity, longitudinal forces, bolus viscosity, and bolus volume. These provide initial insights of practical value for formulators of gel delivery vehicles for anti-HIV microbicidal formulations. PMID- 19547724 TI - Unsteady propagation of a liquid plug in a liquid-lined straight tube. AB - This paper considers the propagation of a liquid plug driven by a constant pressure within a rigid axisymmetric tube whose inner surface is coated by a thin liquid film. The Navier-Stokes equations are solved using the finite-volume method and the SIMPLEST algorithm. The effects of precursor film thickness, initial plug length, pressure drop across the plug, and constant surface tension on the plug behavior and tube wall mechanical stresses are investigated. As a plug propagates through a liquid-lined tube, the plug gains liquid from the leading front film, and it deposits liquid into the trailing film. If the trailing film is thicker (thinner) than the precursor film, the plug volume decreases (increases) as it propagates. For a decreasing volume, eventually the plug ruptures. Under a specific set of conditions, the trailing film thickness equals the precursor film thickness, which leads to steady state results. The plug speed decreases as the precursor film thins because the resistance to the moving front meniscus increases. As the pressure drop across the plug decreases, the plug speed decreases resulting in thinning of the trailing film. As the plug length becomes longer, the viscous resistance in the plug core region increases, which slows the plug and causes the trailing film to become even thinner. The magnitude of the pressure and shear stress at the tube inner wall is maximum in the front meniscus region, and it increases with a thinner precursor film. As the surface tension increases, the plug propagation speed decreases, the strength of the wall pressure in the front meniscus region increases, and the pressure gradient around the peak pressure becomes steeper. PMID- 19547725 TI - Statistical equilibrium of bubble oscillations in dilute bubbly flows. AB - The problem of predicting the moments of the distribution of bubble radius in bubbly flows is considered. The particular case where bubble oscillations occur due to a rapid (impulsive or step change) change in pressure is analyzed, and it is mathematically shown that in this case, inviscid bubble oscillations reach a stationary statistical equilibrium, whereby phase cancellations among bubbles with different sizes lead to time-invariant values of the statistics. It is also shown that at statistical equilibrium, moments of the bubble radius may be computed using the period-averaged bubble radius in place of the instantaneous one. For sufficiently broad distributions of bubble equilibrium (or initial) radius, it is demonstrated that bubble statistics reach equilibrium on a time scale that is fast compared to physical damping of bubble oscillations due to viscosity, heat transfer, and liquid compressibility. The period-averaged bubble radius may then be used to predict the slow changes in the moments caused by the damping. A benefit is that period averaging gives a much smoother integrand, and accurate statistics can be obtained by tracking as few as five bubbles from the broad distribution. The period-averaged formula may therefore prove useful in reducing computational effort in models of dilute bubbly flow wherein bubbles are forced by shock waves or other rapid pressure changes, for which, at present, the strong effects caused by a distribution in bubble size can only be accurately predicted by tracking thousands of bubbles. Some challenges associated with extending the results to more general (nonimpulsive) forcing and strong two-way coupled bubbly flows are briefly discussed. PMID- 19547726 TI - [Not Available]. AB - AIM: We aimed to report our postoperative results in elderly patients that had off-pump coronary bypass grafting. METHOD: Data of 173 patients with isolated coronary bypass 70 years of age or older were retrospectively evaluated. One month follow-up data were evaluated in all patients. After getting verbal informed consent a total of 91 patients were included in the quality of life assessment via EuroQoL form. The data of patients with age of 70-74 and 75 or older were compared. RESULTS: The mean ages of patients at age of 70-74 and >=75 were 71.69+/-0.16 and 76.81+/-0.23 years; respectively. Eleven cases had in hospital mortality (%6 of 173 patients). The mean follow-up period for the group that were reached for EuroQoL assessment was 46,3 +/- 20,8 months. The percent of cases among the whole study group that had participated in the quality of life questionnaire were in good condition in terms of mobility, self-care, usual activities, pain/discomfort, anxiety/depression were %75, %87, %81, %92 and %89; respectively. DISCUSSION: Off-pump coronary bypass operation may be safely used in elderly patients with high quality of life and low morbidity and mortality. PMID- 19547728 TI - [Not Available]. AB - Percutaneous endoscopic-guided gastrostomy (PEG) is done routinely on patients who suffer from inability to feed by mouth. PEG is generally considered a safe procedure with a low complication rate. A commonly underreported complication of PEG is malposition. This manuscript is a guideline to diagnosis and management of PEG malposition. We describe the different types of malposition, their diagnosis and management. PMID- 19547729 TI - Stable therapeutic effects thanks to psychodynamic therapy. PMID- 19547727 TI - Comparison of prognostic gene profiles using qRT-PCR in paraffin samples: a retrospective study in patients with early breast cancer. AB - INTRODUCTION: Gene profiling may improve prognostic accuracy in patients with early breast cancer, but this technology is not widely available. We used commercial assays for qRT-PCR to assess the performance of the gene profiles included in the 70-Gene Signature, the Recurrence Score and the Two-Gene Ratio. METHODS: 153 patients with early breast cancer and a minimum follow-up of 5 years were included. All tumours were positive for hormonal receptors and 38% had positive lymph nodes; 64% of patients received adjuvant chemotherapy. RNA was extracted from formalin-fixed paraffin-embedded (FFPE) specimens using a specific kit. qRT-PCR amplifications were performed with TaqMan Gene Expression Assays products. We applied the three gene-expression-based models to our patient cohort to compare the predictions derived from these gene sets. RESULTS: After a median follow-up of 91 months, 22% of patients relapsed. The distant metastasis-free survival (DMFS) at 5 years was calculated for each profile. For the 70-Gene Signature, DMFS was 95% -good prognosis- versus 66% -poor prognosis. In the case of the Recurrence Score, DMFS was 98%, 81% and 69% for low, intermediate and high risk groups, respectively. Finally, for the Two-Gene Ratio, DMFS was 86% versus 70%. The 70-Gene Signature and the Recurrence Score were highly informative in identifying patients with distant metastasis, even in multivariate analysis. CONCLUSION: Commercially available assays for qRT-PCR can be used to assess the prognostic utility of previously published gene expression profiles in FFPE material from patients with early breast cancer. Our results, with the use of a different platform and with different material, confirm the robustness of the 70 Gene Signature and represent an independent test for the Recurrence Score, using different primer/probe sets. PMID- 19547730 TI - Opipramol as nighttime medication. PMID- 19547732 TI - Binge drinking in childhood and adolescence: epidemiology, consequences, and interventions. AB - BACKGROUND: Episodic excessive alcohol consumption ("binge drinking") among children and adolescents has become a serious public health problem in Germany and is associated with a variety of risks. METHODS: Selective literature search of the Ovid Medline database from 1998 to 2008. RESULTS: Episodic excessive alcohol consumption is associated not only with somatic complications, but also with traffic accidents and other types of accident, violent behavior, and suicide. The more frequently a child or adolescent drinks to excess, and the younger he or she is, the greater is the risk of developing an alcohol-related disorder (alcohol misuse or dependence syndrome). In the USA, brief motivational interventions have been shown to have a small to medium-sized beneficial effect in reducing further binge drinking and its complications. CONCLUSIONS: The intervention HaLT ("Stop," also an acronym for Hart am Limit--"near the limit") is performed in a number of regions in Germany. Further types of brief motivating intervention should be developed and evaluated to prevent the development of alcohol-related disorders, where indicated, in children and adolescents that engage in binge drinking. PMID- 19547733 TI - Cowpox virus infection in pet rat owners: not always immediately recognized. AB - BACKGROUND: The aim of this article is to make physicians of all specialties aware of the possible variations of clinical course in human cowpox infection. This has been a matter of current interest since the detection of a first cluster of infections among owners of white pet rats in the Krefeld area in the spring of 2008. Two further cases arose in the Krefeld area in November 2008, and there have since been multiple further reports from various regions in Germany and the neighboring countries. METHOD: The authors report on the first six documented cases of infection with cowpox virus among young persons owning pet rats, with both typical and atypical clinical courses. RESULTS: The clinical, molecular biological, and serological findings confirmed cowpox virus infection in all six cases. The DNA sequence of the cowpox virus hemagglutinin gene was identical in all patients. The infections had arisen after direct contact with pet rats. CONCLUSIONS: Molecular genetic analysis of the cases described here suggests that the observed occurrence of cowpox virus infection among human beings and pet rats in multiple geographical areas represents a unitary epidemiological event that has not yet come under control. Further cases can be expected. PMID- 19547734 TI - Confidence interval or p-value?: part 4 of a series on evaluation of scientific publications. AB - BACKGROUND: An understanding of p-values and confidence intervals is necessary for the evaluation of scientific articles. This article will inform the reader of the meaning and interpretation of these two statistical concepts. METHODS: The uses of these two statistical concepts and the differences between them are discussed on the basis of a selective literature search concerning the methods employed in scientific articles. RESULTS/CONCLUSIONS: P-values in scientific studies are used to determine whether a null hypothesis formulated before the performance of the study is to be accepted or rejected. In exploratory studies, p values enable the recognition of any statistically noteworthy findings. Confidence intervals provide information about a range in which the true value lies with a certain degree of probability, as well as about the direction and strength of the demonstrated effect. This enables conclusions to be drawn about the statistical plausibility and clinical relevance of the study findings. It is often useful for both statistical measures to be reported in scientific articles, because they provide complementary types of information. PMID- 19547735 TI - Expertise is crucial. PMID- 19547737 TI - Knock-out drugs: their prevalence, modes of action, and means of detection. AB - BACKGROUND: Knock-out drugs are used to facilitate the commission of a crime, generally either robbery or sexual assault. Although media reports on the use of knock-out drugs have become more frequent, there are no robust epidemiological data on the incidence of drug-facilitated robbery or sexual assault, presumably because many crimes of these types do not enter into official statistics. METHODS: The authors describe the modes of action and toxicological means of detection of the substances most frequently used as knock-out drugs on the basis of a selective literature research on the terms "drug-facilitated sexual assaults" (DFSA) and "drug-facilitated crimes" (DFC). RESULTS: The most frequently used drug in cases of sexual assault is still alcohol (ca. 40% to 60%), followed by illegal drugs (cannabis, cocaine). The presence of involuntarily consumed medications and drugs of abuse is demonstrated by routine toxicological analysis only in relatively few cases (ca. 2%). The substances most commonly found are benzodiazepines, followed by other hypnotics. In Europe, the illegal substance gamma-hydroxybutyric acid (GHB, "Liquid Ecstasy"), often mentioned as a "date-rape drug," is only rarely detected with sufficient medicolegal certainty. This may be due to its rapid elimination (it is detectable in blood for up to 8 hours, in urine for up to 12 hours) as well as its physiological occurrence in the body. If the toxicological analysis of blood and urine is negative in a case of suspected DFSA, then the analysis of a hair sample about four weeks after the assault can detect the presence of drugs consumed at that time. If the victim has long hair, it may be possible to detect knock-out drugs taken more than four weeks earlier. In Europe, convictions for drug facilitated crimes are comparatively rare, mainly because of the difficulty of demonstrating conclusive evidence. CONCLUSIONS: A careful medical history and physical examination and the careful taking of biological samples for toxicological analysis form the basis for the detection of drug-facilitated crimes. PMID- 19547739 TI - Universal auditory screening of neonates and hearing disorders in childhood. PMID- 19547738 TI - Typhus and other rickettsioses: emerging infections in Germany. AB - BACKGROUND: Rickettsioses are diseases caused by rickettsiae, obligate intracellular bacteria that are transmitted by arthropods to humans. They cause various types of spotted fever and typhus. METHODS: A review of the literature is presented along with the authors' own findings. RESULTS: Six indigenous species of rickettsiae have been found in Germany to date, five of which have been described as human pathogens in other countries. Rickettsia slovaca causes tick borne lymphadenitis (TIBOLA). Rickettsia helvetica is a known pathogen of nonspecific fever; its role in endocarditis is still under investigation. Rickettsia felis causes so-called flea-borne spotted fever. Rickettsia monacensis and Rickettsia massiliae were recently shown to cause the classical form of tick borne spotted fever. The sixth indigenous species in Germany, Rickettsia sp. RpA4, has not yet been associated with any human disease. The most important rickettsioses imported to Germany by travelers are African tick bite fever and Mediterranean spotted fever. CONCLUSIONS: Modern molecular biological techniques have enabled the detection of a number of rickettsial species in Germany. The medical importance of these illnesses in Germany remains to be determined. In travel medicine, imported rickettsioses play a role that should not be underestimated. PMID- 19547740 TI - Support for general practitioners is uncertain. PMID- 19547742 TI - Positional cues in the Drosophila nerve cord: semaphorins pattern the dorso ventral axis. AB - During the development of neural circuitry, neurons of different kinds establish specific synaptic connections by selecting appropriate targets from large numbers of alternatives. The range of alternative targets is reduced by well organised patterns of growth, termination, and branching that deliver the terminals of appropriate pre- and postsynaptic partners to restricted volumes of the developing nervous system. We use the axons of embryonic Drosophila sensory neurons as a model system in which to study the way in which growing neurons are guided to terminate in specific volumes of the developing nervous system. The mediolateral positions of sensory arbors are controlled by the response of Robo receptors to a Slit gradient. Here we make a genetic analysis of factors regulating position in the dorso-ventral axis. We find that dorso-ventral layers of neuropile contain different levels and combinations of Semaphorins. We demonstrate the existence of a central to dorsal and central to ventral gradient of Sema 2a, perpendicular to the Slit gradient. We show that a combination of Plexin A (Plex A) and Plexin B (Plex B) receptors specifies the ventral projection of sensory neurons by responding to high concentrations of Semaphorin 1a (Sema 1a) and Semaphorin 2a (Sema 2a). Together our findings support the idea that axons are delivered to particular regions of the neuropile by their responses to systems of positional cues in each dimension. PMID- 19547743 TI - Management effectiveness of the world's marine fisheries. AB - Ongoing declines in production of the world's fisheries may have serious ecological and socioeconomic consequences. As a result, a number of international efforts have sought to improve management and prevent overexploitation, while helping to maintain biodiversity and a sustainable food supply. Although these initiatives have received broad acceptance, the extent to which corrective measures have been implemented and are effective remains largely unknown. We used a survey approach, validated with empirical data, and enquiries to over 13,000 fisheries experts (of which 1,188 responded) to assess the current effectiveness of fisheries management regimes worldwide; for each of those regimes, we also calculated the probable sustainability of reported catches to determine how management affects fisheries sustainability. Our survey shows that 7% of all coastal states undergo rigorous scientific assessment for the generation of management policies, 1.4% also have a participatory and transparent processes to convert scientific recommendations into policy, and 0.95% also provide for robust mechanisms to ensure the compliance with regulations; none is also free of the effects of excess fishing capacity, subsidies, or access to foreign fishing. A comparison of fisheries management attributes with the sustainability of reported fisheries catches indicated that the conversion of scientific advice into policy, through a participatory and transparent process, is at the core of achieving fisheries sustainability, regardless of other attributes of the fisheries. Our results illustrate the great vulnerability of the world's fisheries and the urgent need to meet well-identified guidelines for sustainable management; they also provide a baseline against which future changes can be quantified. PMID- 19547744 TI - Evolution of phosphoregulation: comparison of phosphorylation patterns across yeast species. AB - The extent by which different cellular components generate phenotypic diversity is an ongoing debate in evolutionary biology that is yet to be addressed by quantitative comparative studies. We conducted an in vivo mass-spectrometry study of the phosphoproteomes of three yeast species (Saccharomyces cerevisiae, Candida albicans, and Schizosaccharomyces pombe) in order to quantify the evolutionary rate of change of phosphorylation. We estimate that kinase-substrate interactions change, at most, two orders of magnitude more slowly than transcription factor (TF)-promoter interactions. Our computational analysis linking kinases to putative substrates recapitulates known phosphoregulation events and provides putative evolutionary histories for the kinase regulation of protein complexes across 11 yeast species. To validate these trends, we used the E-MAP approach to analyze over 2,000 quantitative genetic interactions in S. cerevisiae and Sc. pombe, which demonstrated that protein kinases, and to a greater extent TFs, show lower than average conservation of genetic interactions. We propose therefore that protein kinases are an important source of phenotypic diversity. PMID- 19547745 TI - Light, sleep, and circadian rhythms: together again. PMID- 19547746 TI - Self-organization of the Escherichia coli chemotaxis network imaged with super resolution light microscopy. AB - The Escherichia coli chemotaxis network is a model system for biological signal processing. In E. coli, transmembrane receptors responsible for signal transduction assemble into large clusters containing several thousand proteins. These sensory clusters have been observed at cell poles and future division sites. Despite extensive study, it remains unclear how chemotaxis clusters form, what controls cluster size and density, and how the cellular location of clusters is robustly maintained in growing and dividing cells. Here, we use photoactivated localization microscopy (PALM) to map the cellular locations of three proteins central to bacterial chemotaxis (the Tar receptor, CheY, and CheW) with a precision of 15 nm. We find that cluster sizes are approximately exponentially distributed, with no characteristic cluster size. One-third of Tar receptors are part of smaller lateral clusters and not of the large polar clusters. Analysis of the relative cellular locations of 1.1 million individual proteins (from 326 cells) suggests that clusters form via stochastic self-assembly. The super resolution PALM maps of E. coli receptors support the notion that stochastic self assembly can create and maintain approximately periodic structures in biological membranes, without direct cytoskeletal involvement or active transport. PMID- 19547747 TI - Prevalence of Buruli ulcer in Akonolinga health district, Cameroon: results of a cross sectional survey. AB - BACKGROUND: Buruli ulcer (BU) is a chronic, indolent necrotizing disease of the skin and underlying tissues caused by Mycobacterium ulcerans, which may result in functional incapacity. In 2002, Medecins Sans Frontieres (MSF) opened a BU programme in Akonolinga Hospital, Cameroon, offering antibiotic treatment, surgery and general medical care. Six hundred patients have been treated in the project to date. However, due to the nature of the disease and its stigmatization, determining the exact prevalence and burden of disease is difficult and current estimates may not reflect the magnitude of the problem. The objectives of this survey were to estimate the prevalence of BU in the health district of Akonolinga, describe the geographic extension of the highly endemic area within the health district, and determine the programme coverage and its geographical distribution. METHODOLOGY/PRINCIPAL FINDINGS: We conducted a cross sectional population survey using centric systematic area sampling (CSAS). A 15x15 km grid (quadrats of 225 km(2)) was overlaid on a map of Akonolinga district with its position chosen to maximize the area covered by the survey. Quadrats were selected if more than 50% of the quadrat was inside of the health district. The chiefdom located closest to the centre of each quadrat was selected and Buruli cases were identified using an active case finding strategy (the sensitivity of the strategy was estimated by capture-recapture). WHO-case definitions were used for nodules, plaque, ulcer, oedema and sequelae. Out of a total population of 103,000 inhabitants, 26,679 were surveyed within the twenty quadrats. Sensitivity of the case finding strategy was estimated to be 84% (95%CI 54-97%). The overall prevalence was 0.47% (n = 105) for all cases including sequelae and 0.25% (n = 56) for active stages of the disease. Five quadrats had a high prevalence of >0.6% to 0.9%, 5 a prevalence >0.3% to 0.6% and 10 quadrats <0.3%. The quadrats with the high prevalence were situated along the rivers Nyong and Mfoumou. Overall coverage of the project was 18% (12-27%) for all cases and 16% (9-18%) for active cases, but was limited to the quadrats neighbouring Akonolinga Hospital. CONCLUSIONS/SIGNIFICANCE: Prevalence was highest in the area neighbouring the Nyong River. Coverage was limited to the area close to the hospital and efforts have to be made to increase access to care in the high prevalence areas. Use of the CSAS method was particularly useful for project planning and to identify priority areas of intervention. An added benefit of the method is that the survey procedure incorporated an awareness campaign, providing information about the disease and treatment to the population. PMID- 19547748 TI - Leptospira interrogans stably infects zebrafish embryos, altering phagocyte behavior and homing to specific tissues. AB - Leptospirosis is an extremely widespread zoonotic infection with outcomes ranging from subclinical infection to fatal Weil's syndrome. Despite the global impact of the disease, key aspects of its pathogenesis remain unclear. To examine in detail the earliest steps in the host response to leptospires, we used fluorescently labelled Leptospira interrogans serovar Copenhageni to infect 30 hour post fertilization zebrafish embryos by either the caudal vein or hindbrain ventricle. These embryos have functional innate immunity but have not yet developed an adaptive immune system. Furthermore, they are optically transparent, allowing direct visualization of host-pathogen interactions from the moment of infection. We observed rapid uptake of leptospires by phagocytes, followed by persistent, intracellular infection over the first 48 hours. Phagocytosis of leptospires occasionally resulted in formation of large cellular vesicles consistent with apoptotic bodies. By 24 hours, clusters of infected phagocytes were accumulating lateral to the dorsal artery, presumably in early hematopoietic tissue. Our observations suggest that phagocytosis may be a key defense mechanism in the early stages of leptospirosis, and that phagocytic cells play roles in immunopathogenesis and likely in the dissemination of leptospires to specific target tissues. PMID- 19547749 TI - High burden of impetigo and scabies in a tropical country. AB - BACKGROUND: Impetigo and scabies are endemic diseases in many tropical countries; however the epidemiology of these diseases is poorly understood in many areas, particularly in the Pacific. METHODOLOGY/PRINCIPAL FINDINGS: We conducted three epidemiological studies in 2006 and 2007 to determine the burden of disease due to impetigo and scabies in children in Fiji using simple and easily reproducible methodology. Two studies were performed in primary school children (one study was a cross-sectional study and the other a prospective cohort study over ten months) and one study was performed in infants (cross-sectional). The prevalence of active impetigo was 25.6% (95% CI 24.1-27.1) in primary school children and 12.2% (95% CI 9.3-15.6) in infants. The prevalence of scabies was 18.5% (95% CI 17.2 19.8) in primary school children and 14.0% (95% CI 10.8-17.2) in infants. The incidence density of active impetigo, group A streptococcal (GAS) impetigo, Staphylococcus aureus impetigo and scabies was 122, 80, 64 and 51 cases per 100 child-years respectively. Impetigo was strongly associated with scabies infestation (odds ratio, OR, 2.4, 95% CI 1.6-3.7) and was more common in Indigenous Fijian children when compared with children of other ethnicities (OR 3.6, 95% CI 2.7-4.7). The majority of cases of active impetigo in the children in our study were caused by GAS. S. aureus was also a common cause (57.4% in school aged children and 69% in infants). CONCLUSIONS/SIGNIFICANCE: These data suggest that the impetigo and scabies disease burden in children in Fiji has been underestimated, and possibly other tropical developing countries in the Pacific. These diseases are more than benign nuisance diseases and consideration needs to be given to expanded public health initiatives to improve their control. PMID- 19547750 TI - MIRO and IRbase: IT tools for the epidemiological monitoring of insecticide resistance in mosquito disease vectors. AB - BACKGROUND: Monitoring of insect vector populations with respect to their susceptibility to one or more insecticides is a crucial element of the strategies used for the control of arthropod-borne diseases. This management task can nowadays be achieved more efficiently when assisted by IT (Information Technology) tools, ranging from modern integrated databases to GIS (Geographic Information System). Here we describe an application ontology that we developed de novo, and a specially designed database that, based on this ontology, can be used for the purpose of controlling mosquitoes and, thus, the diseases that they transmit. METHODOLOGY/PRINCIPAL FINDINGS: The ontology, named MIRO for Mosquito Insecticide Resistance Ontology, developed using the OBO-Edit software, describes all pertinent aspects of insecticide resistance, including specific methodology and mode of action. MIRO, then, forms the basis for the design and development of a dedicated database, IRbase, constructed using open source software, which can be used to retrieve data on mosquito populations in a temporally and spatially separate way, as well as to map the output using a Google Earth interface. The dependency of the database on the MIRO allows for a rational and efficient hierarchical search possibility. CONCLUSIONS/SIGNIFICANCE: The fact that the MIRO complies with the rules set forward by the OBO (Open Biomedical Ontologies) Foundry introduces cross-referencing with other biomedical ontologies and, thus, both MIRO and IRbase are suitable as parts of future comprehensive surveillance tools and decision support systems that will be used for the control of vector borne diseases. MIRO is downloadable from and IRbase is accessible at VectorBase, the NIAID-sponsored open access database for arthropod vectors of disease. PMID- 19547751 TI - The differential diagnosis of food intolerance. AB - INTRODUCTION: More than 20% of the population in industrialized countries suffer from food intolerance or food allergy. METHODS: Selective literature search for relevant publications in PubMed and the Cochrane Library combined with further data from the interdisciplinary database on chronic inflammatory and allergic diseases of the Erlangen University Hospital. RESULTS: The majority of cases of food intolerance (15% to 20%) are due to non-immunological causes. These causes range from pseudoallergic reactions to enzymopathies, chronic infections, and psychosomatic reactions that are associated with food intolerance. The prevalence of true food allergy, i.e., immunologically mediated intolerance reactions, is only 2% to 5%. CONCLUSIONS: The differential diagnosis of food intolerance is broad. Therefore, a structured diagnostic algorithm with input from multiple clinical disciplines should be applied. The treatment consists of eliminating the offending substance from the diet as well as medications and psychosomatic support, when indicated. PMID- 19547752 TI - Human telomere length correlates to the size of the associated chromosome arm. AB - The majority of human telomere length studies have focused on the overall length of telomeres within a cell. In fact, very few studies have examined telomere length for individual chromosome arms. The objective of this study was to examine the relationship between chromosome arm size and the relative length of the associated telomere. Quantitative Fluorescence In Situ Hybridization (Q-FISH) was used to measure the relative telomere length of each chromosome arm in metaphases from cultured lymphocytes of 17 individuals. A statistically significant positive correlation (r = 0.6) was found between telomere length and the size of the associated chromosome arm, which was estimated based on megabase pair measurements from (http://www.ncbi.nlm.nih.gov/projects/mapview/). PMID- 19547754 TI - Long-term potentiation in isolated dendritic spines. AB - BACKGROUND: In brain, N-methyl-D-aspartate (NMDA) receptor (NMDAR) activation can induce long-lasting changes in synaptic alpha-amino-3-hydroxy-5-methylisoxazole-4 propionate (AMPA) receptor (AMPAR) levels. These changes are believed to underlie the expression of several forms of synaptic plasticity, including long-term potentiation (LTP). Such plasticity is generally believed to reflect the regulated trafficking of AMPARs within dendritic spines. However, recent work suggests that the movement of molecules and organelles between the spine and the adjacent dendritic shaft can critically influence synaptic plasticity. To determine whether such movement is strictly required for plasticity, we have developed a novel system to examine AMPAR trafficking in brain synaptosomes, consisting of isolated and apposed pre- and postsynaptic elements. METHODOLOGY/PRINCIPAL FINDINGS: We report here that synaptosomes can undergo LTP like plasticity in response to stimuli that mimic synaptic NMDAR activation. Indeed, KCl-evoked release of endogenous glutamate from presynaptic terminals, in the presence of the NMDAR co-agonist glycine, leads to a long-lasting increase in surface AMPAR levels, as measured by [(3)H]-AMPA binding; the increase is prevented by an NMDAR antagonist 2-amino-5-phosphonopentanoic acid (AP5). Importantly, we observe an increase in the levels of GluR1 and GluR2 AMPAR subunits in the postsynaptic density (PSD) fraction, without changes in total AMPAR levels, consistent with the trafficking of AMPARs from internal synaptosomal compartments into synaptic sites. This plasticity is reversible, as the application of AMPA after LTP depotentiates synaptosomes. Moreover, depotentiation requires proteasome-dependent protein degradation. CONCLUSIONS/SIGNIFICANCE: Together, the results indicate that the minimal machinery required for LTP is present and functions locally within isolated dendritic spines. PMID- 19547753 TI - BDNF activates mTOR to regulate GluR1 expression required for memory formation. AB - BACKGROUND: The mammalian target of Rapamycin (mTOR) kinase plays a key role in translational control of a subset of mRNAs through regulation of its initiation step. In neurons, mTOR is present at the synaptic region, where it modulates the activity-dependent expression of locally-translated proteins independently of mRNA synthesis. Indeed, mTOR is necessary for different forms of synaptic plasticity and long-term memory (LTM) formation. However, little is known about the time course of mTOR activation and the extracellular signals governing this process or the identity of the proteins whose translation is regulated by this kinase, during mnemonic processing. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that consolidation of inhibitory avoidance (IA) LTM entails mTOR activation in the dorsal hippocampus at the moment of and 3 h after training and is associated with a rapid and rapamycin-sensitive increase in AMPA receptor GluR1 subunit expression, which was also blocked by intra-hippocampal delivery of GluR1 antisense oligonucleotides (ASO). In addition, we found that pre- or post training administration of function-blocking anti-BDNF antibodies into dorsal CA1 hampered IA LTM retention, abolished the learning-induced biphasic activation of mTOR and its readout, p70S6K and blocked GluR1 expression, indicating that BDNF is an upstream factor controlling mTOR signaling during fear-memory consolidation. Interestingly, BDNF ASO hindered LTM retention only when given into dorsal CA1 1 h after but not 2 h before training, suggesting that BDNF controls the biphasic requirement of mTOR during LTM consolidation through different mechanisms: an early one involving BDNF already available at the moment of training, and a late one, happening around 3 h post-training that needs de novo synthesis of this neurotrophin. CONCLUSIONS/SIGNIFICANCE: IN CONCLUSION, OUR FINDINGS DEMONSTRATE THAT: 1) mTOR-mediated mRNA translation is required for memory consolidation during at least two restricted time windows; 2) this kinase acts downstream BDNF in the hippocampus and; 3) it controls the increase of synaptic GluR1 necessary for memory consolidation. PMID- 19547755 TI - Increased sensitivity to thermal pain following a single opiate dose is influenced by the COMT val(158)met polymorphism. AB - Increased pain sensitivity after opioid administration (opioid-induced hyperalgesia) and/or repeated painful stimuli is an individually varying and clinically important phenomenon. The functional polymorphism (val(158)met) of the Catechol-O-methyltransferase (COMT) gene regulates the metabolism of dopamine/noradrenaline. Individuals homozygous for the met(158) allele have been reported to have increased pain sensitivity and there are findings of lower micro opioid system activation during sustained pain. We hypothesized that met/met individuals would exhibit higher pain sensitization and opioid-induced hyperalgesia in response to repeated pain stimuli and an intravenous injection of an opioid drug. Participants were 43 healthy subjects who went through an experiment where five blocks of pain were induced to the hand using a heat probe. After each stimulus subjects rated the pain on a visual analogue scale (VAS) from 0 mm (no pain) to 100 mm (worst possible pain). Before the second stimulus there was an intravenous injection of a rapid and potent opioid drug. At baseline there was no difference in pain ratings between the COMTval(158)met genotypes, F(2, 39)<1. However, a repeated measures ANOVA for all five stimuli revealed a main effect for COMTval(158)met genotype, F(2, 36) = 4.17, p = 0.024. Met/met individuals reported significantly more pain compared to val/val, p = 0.010. A pairwise comparison of baseline and the opioid intervention demonstrated that analgesia was induced in all groups (p = 0.042) without a separating effect for genotype (n.s). We suggest that the initial response of the descending pain system is not influenced by the COMTval(158)met polymorphism but when the system is challenged the difference is revealed. An important clinical implication of this may be that the COMTval(158)met related differences may be more expressed in individuals where the inhibitory system is already challenged and sensitive, e.g. chronic pain patients. This has to be proven in future studies where the impact of the COMTval(158)met polymorphism on opioid treatment in patients is addressed. PMID- 19547756 TI - Impaired interleukin-1beta and c-Fos expression in the hippocampus is associated with a spatial memory deficit in P2X(7) receptor-deficient mice. AB - Recent evidence suggests that interleukin-1beta (IL-1beta), which was originally identified as a proinflammatory cytokine, is also required in the brain for memory processes. We have previously shown that IL-1beta synthesis in the hippocampus is dependent on P2X(7) receptor (P2X(7)R), which is an ionotropic receptor of ATP. To substantiate the role of P2X(7)R in both brain IL-1beta expression and memory processes, we examined the induction of IL-1beta mRNA expression in the hippocampus of wild-type (WT) and homozygous P2X(7) receptor knockout mice (P2X(7)R(-/-)) following a spatial memory task. The spatial recognition task induced both IL-1beta mRNA expression and c-Fos protein activation in the hippocampus of WT but not of P2X(7)R(-/-) mice. Remarkably, P2X(7)R(-/-) mice displayed spatial memory impairment in a hippocampal-dependant task, while their performances in an object recognition task were unaltered. Taken together, our results show that P2X(7)R plays a critical role in spatial memory processes and the associated hippocampal IL-1beta mRNA synthesis and c-Fos activation. PMID- 19547757 TI - Pilot study of essential drug quality in two major cities in India. AB - BACKGROUND: India is an increasingly influential player in the global pharmaceutical market. Key parts of the drug regulatory system are controlled by the states, each of which applies its own standards for enforcement, not always consistent with others. A pilot study was conducted in two major cities in India, Delhi and Chennai, to explore the question/hypothesis/extent of substandard and counterfeit drugs available in the market and to discuss how the Indian state and federal governments could improve drug regulation and more importantly regulatory enforcement to combat these drugs. METHODOLOGY/PRINCIPAL FINDINGS: Random samples of antimalarial, antibiotic, and antimycobacterial drugs were collected from pharmacies in urban and peri-urban areas of Delhi and Chennai, India. Semi quantitative thin-layer chromatography and disintegration testing were used to measure the concentration of active ingredients against internationally acceptable standards. 12% of all samples tested from Delhi failed either one or both tests, and were substandard. 5% of all samples tested from Chennai failed either one or both tests, and were substandard. Spatial heterogeneity between pharmacies was observed, with some having more or less substandard drugs (30% and 0% respectively), as was product heterogeneity, with some drugs being more or less frequently substandard (12% and 7% respectively). CONCLUSIONS/SIGNIFICANCE: In a study using basic field-deployable techniques of lesser sensitivity rather than the most advanced laboratory-based techniques, the prevalence of substandard drugs in Delhi and Chennai is confirmed to be roughly in accordance with the Indian government's current estimates. However, important spatial and product heterogeneity exists, which suggests that India's substandard drug problem is not ubiquitous, but driven by a subset of manufacturers and pharmacies which thrive in an inadequately regulated environment. It is likely that the drug regulatory system in India needs to be improved for domestic consumption, and because India is an increasingly important exporter of drugs for both developed and developing countries. Some poor countries with high burdens of disease have weak drug regulatory systems and import many HIV/AIDS, tuberculosis and malaria drugs from India. PMID- 19547758 TI - [Not Available]. AB - BACKGROUND: There is a noticeable decline in the number of patients who undergo coronary artery revascularization procedures. The change is definite as it is reported by many centers around the world. This trend is of great concern to cardiac surgeons because of its impact on their practice, its adverse effect on training and the degree of uncertainty it throws into future of the specialty. METHODS: The data of the cardiac catheterization laboratory at the King Fahad Cardiac Center in the period between 1986 and 2006 was examined looking at the changing pattern of management of patients who undergo cardiac catheterization. RESULTS: In the early years, angioplasty was attempted in around 10% of patients leaving the rest for surgical consideration or medical therapy. Currently only 15% of patients who undergo selective coronary angiography are referred for surgery. The majority are offered angioplasty and stenting. The trend is towards more catheter-based interventions and less towards surgery. CONCLUSIONS: Our findings are in agreement with the general consensus about the specialty. Cardiac surgeons should perhaps consider acquiring new skills which may be outside the operating room. Adding catheter based intervention particularly in valves to cardiac surgery training would be a bonus for the future surgeons that will give them the necessary edge to meet the new challenges. It is incumbent on the leaders in the field to establish a clear strategy for the future. MINI-ABSTRACT: Impact of advances in invasive cardiology on cardiac surgery, based on actual analysis of the pattern of referrals to surgery of over two decades. PMID- 19547759 TI - Diminished expression of ICOS, GITR and CTLA-4 at the mRNA level in T regulatory cells of children with newly diagnosed type 1 diabetes. AB - Diabetes mellitus is one of the most common chronic diseases in children. T regulatory cells (Tregs) modulate response to autoantigens and probably play a role in pathogenesis of type 1 diabetes (T1DM). The aim of the present study was the assessment of T regulatory cells including their percentages and expression of critical genes in these cells in children with newly diagnosed type 1 diabetes. The examined group consisted of 50 children with T1DM. A flow cytometric analysis of T-cell subpopulations was performed using the following markers: anti-CD4, anti-CD25 and anti-CD127 (=IL-7R). Additionally, T regulatory cells were isolated for assessment of mRNA levels for chosen genes with the real time RT-PCR technique. The percentages of CD4(+)CD25(high)CD127(dim/-) were very low and did not differ between T1DM and control children. We did not observe any statistically significant differences between healthy and diabetic children in mRNA expression for FoxP3, IL-7R (CD127), IL-8RA, IL-10RA, IL-12A, IL-2RA (CD25), IL-21, STAT1, STAT3, SOCS2, SOCS3, TGF-beta1-R1, TGF-beta-R2 and TBX-21 genes. Interestingly the mRNA level for CTLA-4, ICOS1, IL-23, IL-27, SMAD3 and GITR were lower in Treg cells of children with diabetes compared to the control patients. No disturbances in the percentages of T regulatory cells in patients with diabetes but diminished expression of some elements important in Treg function could be the result of an immunologic imbalance accompanying the onset of the diabetes. The results of our study should be used in future research in the field of immunotherapy in pediatric diabetes. PMID- 19547760 TI - Does routine pain assessment result in better care? AB - BACKGROUND: Although a variety of national organizations such as the Canadian Pain Society, the American Pain Society and the Joint Commission on Accreditation of Health Care Organizations have advanced the idea that pain should be assessed on a routine basis, there is little evidence that systematic pain assessment information is used routinely by clinicians even when it is readily available. OBJECTIVE: To determine whether systematic pain assessment information alters medical practitioners' clinical practices. METHODS: A population of seniors with complex medical problems who were evaluated by case coordinators was studied. Case coordinators were assigned to either an experimental or control patient assessment condition. Control condition patients were assessed as usual. In the experimental condition, a psychometrically valid pain assessment battery as well as the Geriatric Depression Scale - Short Form (because depression and chronic pain are frequently comorbid) were integrated into the routine case coordination assessment. A summary of the results of the depression and pain assessments was subsequently sent to physicians via mail and fax. Patients were also given copies of the assessment summaries and were asked to discuss these with their physicians. Physicians' medication prescriptions were monitored over time through the database of the provincial ministry of health. RESULTS: At the end of the study, no significant differences between experimental and control patients were found with respect to medications prescribed or patient self-reports of pain. Nonetheless, there was a significant relationship between Geriatric Depression Scale -- Short Form scores and pain medications prescribed for patients in the experimental condition. Moreover, indexes of overall pain intensity did not change significantly over time. CONCLUSIONS: The findings do not support the idea that the availability of systematic pain assessment information leads to change in clinician's medication practices. As such, educational interventions and public policy initiatives are needed to ensure that treatment providers do not only gather but also use pain assessment information. PMID- 19547762 TI - The influence of addiction risk on nursing students' expectations of patients' pain reports: a clinical vignette approach. AB - OBJECTIVE: To examine the influence of addiction risk (substance abuse history [SAH]) and pain relief (PR) on nursing and non-nursing students' perceptions of pain in a postoperative vignette patient. METHODS: Using a 2 x 2 design, the independent variables SAH (present/+, absent/-) and PR (adequate, little) were varied systematically to produce four vignettes. Participants were randomly assigned to receive one of the four vignettes that described a 45-year-old man after a total hip replacement. Participants rated the vignette patient's experienced and reported pain intensity (PI) on a 0 mm to 100 mm visual analogue scale and addiction risk on a 0 mm to 100 mm visual analogue scale. A pain congruence (PC) score was calculated (PC = reported PI -- experienced PI), and was interpreted as congruent (+/-2 mm) or incongruent (+2 mm to +100 mm for expected pain over-reporting; -2 mm to -100 mm for expected pain under reporting). RESULTS: Responses from undergraduate nursing (n=89) and non-nursing (n=88) students were analyzed. The estimated addiction risk was significantly lower in nursing (14% to 45%) versus non-nursing students (50%). Nursing students' mean PC scores were not significantly altered by SAH alone. Expectations of pain over-reporting were observed under conditions of SAH+/adequate PR, but not SAH+/little PR. In non-nursing students, SAH and PR were significant and independent factors influencing mean PC scores in the direction of pain over-reporting. CONCLUSION: Under most conditions, nursing students expected pain under-reporting by the postoperative vignette patient. However, nursing students did expect pain to be over-reported when addiction risk was high and PR was adequate. These data suggest that nursing students' expectations regarding pain over- and under-reporting were sensitive to perceptions of addiction risk, but involved additional factors (eg, level of PR). PMID- 19547761 TI - Gabapentin decreases morphine consumption and improves functional recovery following total knee arthroplasty. AB - BACKGROUND: Moderate to severe pain after total knee arthroplasty often interferes with postoperative rehabilitation and delays discharge from hospital. The present study examined the effects of a four-day postoperative gabapentin (GBP) regimen versus placebo on opioid consumption, pain scores and knee flexion, as well as adverse effects, after total knee arthroplasty. METHODS: After obtaining research ethics board approval and informed consent, 40 patients were enrolled in a randomized, single-blind, placebo-controlled, open-label study. Patients were assigned to one of five groups - preoperative placebo/postoperative placebo (G1), preoperative GBP 600 mg/postoperative placebo (G2), preoperative GBP 600 mg/postoperative GBP 100 mg three times per day (G3), preoperative GBP 600 mg/ postoperative GBP 200 mg three times per day (G4) and preoperative GBP 600 mg/postoperative GBP 300 mg three times per day (G5). Postoperative GBP or placebo was continued for four days after surgery. Two hours before surgery, all patients received celecoxib 400 mg. Based on the above groupings, patients in G1 received placebo medication, whereas patients in G2, G3, G4 and G5 received gabapentin 600 mg 2 h preoperatively. All patients received femoral and sciatic nerve blocks, followed by spinal anesthesia. Beginning in the postanesthetic care unit, all patients received a regimen of celecoxib 200 mg every 12 h for four days and a patient-controlled morphine analgesia pump for 48 h. RESULTS: Thirty six patients (G1, n=7; G2, n=7; G3, n=8; G4, n=7; G5, n=7) completed the study. Data were analyzed by one-way ANOVA followed by a contrast comparing patients who received postoperative GBP (G3, G4 and G5) (n=22) with patients who received placebo postoperatively (G1 and G2) (n=14). Patients who received GBP postoperatively used significantly less patient-controlled morphine analgesia at 24 h, 36 h and 48 h (P<0.05). The postoperative GBP patients had significantly better active assisted knee flexion on postoperative days 2 and 3, with a trend toward better flexion on postoperative day 4. Patients who received GBP postoperatively reported less pruritus than patients who received placebo. There were no differences in pain scores. CONCLUSIONS: These results support the use of GBP in the acute postoperative period. Further trials are needed to delineate the optimal dose, timing and duration of GBP use following surgery. PMID- 19547763 TI - Anxiety influences children's memory for procedural pain. AB - OBJECTIVE: To examine the effects of temperament and trait anxiety on memory for pain. METHODS: Three dimensions of temperament, as well as trait anxiety, were assessed in 36 children (five to 12 years of age) undergoing dental procedures; after the procedure, the children provided pain ratings. Following a six- to eight-week delay, the children reported how much pain they remembered. RESULTS: Most children (85%) accurately recalled their pain. Temperament had no significant effect, but trait-anxious children showed a greater likelihood of recalling more pain than they initially reported, suggesting that they may negatively distort recollections of painful experiences. CONCLUSIONS: When treating children, in particular trait-anxious children, clinicians should consider what children remember as part of pain management intervention. PMID- 19547764 TI - Are we profiling patients with fibromyalgia? PMID- 19547766 TI - Social protection in Brazil: universalism and targeting in the FHC and Lula administrations. AB - This article analyzes the organization of Brazil's social protection system after the Federal Constitution of 1998 (CF 1988). It also demonstrates that 1988 Federal Constitution favored the institutionalization of universalist public policies. This institutionalization took place amidst conflict with the stabilization goals of the Real Plan. The paper argues that such an institutionalization protected public spending in the social area of the macroeconomic management's minimalist project. It also identifies the implementation of social programs targeting the poor during the 1980's decade. Targeting is an innovation directly associated with the adjustment agenda. It reveals that under the FHC and Lula administrations there was an identical adoption of targeted social programs. The targeting of social protection did not possess power of veto over the universalist proposals arising from the democratization in the 1980's. It demonstrates that the Bolsa Familia Program (Family Grant Program - PBF), the main mark of the Lula administration, is a large scale adaptation of the targeted programs of direct transfer of income in the FHC administration. The combination of universalism and targeting expanded the scope of social policy. However, the significant growth in social public spending has not been producing broad social results, although the poor in Brazil have benefited from the PBF's targeting. PMID- 19547765 TI - Understanding the psychosocial profile of women with fibromyalgia syndrome. AB - Fibromyalgia syndrome (FMS) is a disease with a complex etiology characterized by symptoms of widespread pain and fatigue. FMS is more common in women. Both depression and anxiety have been found to be independently associated with the severity of pain in symptoms of FMS. The goal of the present study was to examine the psychosocial profile of women with FMS and to see how the attributions, perceived social support and cognitive biases of women with FMS are related to internalizing ratings of depression and anxiety. The current study included a sample of women with FMS from a local support group and a control group to examine how women with FMS differed from controls with respect to psychosocial variables, and to determine the relationship between these variables. Women with FMS reported a higher external locus of control, lower levels of adaptive cognitive bias, less perceived family support and lower mood than controls. Correlations between these variables were examined within the FMS group, and it was found that an external locus of control was significantly associated with higher ratings of anxiety and depressed mood. These results suggest that beliefs about locus of control and perceived family support of women with FMS may importantly impact their health outcomes, and that treatments related to locus of control and advocating for family support may considerably improve the quality of life of patients with FMS. PMID- 19547767 TI - Pyrrhic potatoes. Comments on the Institutional rules, macroeconomic constraints and innovation of the Brazilian social protection system in the 1990s and 2000s. PMID- 19547768 TI - Tension between paradigms: notes on social policy in Brazil (1988/2008). PMID- 19547769 TI - Choices and policies. PMID- 19547770 TI - Comments on the article Social protection in Brazil: universalism and targeting in the FHC and Lula administrations. PMID- 19547772 TI - Dilemmas of the institutionalization of social policies in twenty years of the Brazilian Constitution of 1988. AB - The objective is to discuss and analyze some elements of the process of institutionalization of social policies in Brazil after the Constitution of 1988, especially those of social welfare (social security, health and social assistance). It is assumed that this process present hybrids that compromise the results prescribed by the Constitution. From one hand, there are important advances in political and organizational apparatus and in the concept of the social question (treated here through three elements: constitutionalization, scope and expansion). Moreover, obstacles remain, particularly in universalization, financing and quality of services, which are contradictory to the advances achieved. The permanence of these hybrids prevents the fairness and citizenship aimed by the 1988 Constitution. PMID- 19547773 TI - Social protection in Brazil: what has changed in social assistance after the 1988 Constitution. AB - This paper discusses the changes in the social assistance component of social protection in the two decades that followed the 1988 Constitution. It discusses the transformations of social protection and social assistance in the past decades and the processes that in the 1990 s advanced the reform of social assistance policies in Brazil without, however, producing great changes in how services were delivered. By 2000 there had been a great expansion of resources as well as benefits and services coverage in social assistance. This paper argues that conditional cash transfer programs, though created and spread in an international context of restrictive policies, when integrated into Brazil s public and universal system of social assistance, substantively widened the scope of social protection. Some effects of the expansion in the Brazilian social protection system are identified: less income inequality; creation of an institutional capacity in the area of social assistance; the social, political and symbolic meaning of the inclusion of a large segment of the population in a public system of social assistance by means of a provision structure that does not belong to traditional devices of philanthropy and clientelism. PMID- 19547774 TI - Brazilian sanitary reform: dilemmas between the instituing and the institutionalized. AB - The article presents the trajectory of social policies in Brazil and identifies, at each stage, the existing social protection model. It affirms that the Federal Constitution of 1988, when it introduced the concept of Social Security and created the National Health Care System, represented a rupture with what came before. The subordination of the principles of justice and social inclusion, which guided the design of this new social protection model, to a liberal and monetarist policy had important impacts in the phase of implementation of the social policies. However, the dilemma that such policies go through and, in particular, the construction of the National Health Care System must be analyzed from a theoretical perspective that encompasses the agreements and disagreements between the three movements that characterize Brazilian Sanitary Reform, which are: subjectivation, constitutionalization and institutionalization. PMID- 19547775 TI - The Brazilian health system between norms and facts: mitigated universalization and subsidized stratification. AB - This paper will focus on recent changes in the relationships between the public and the private sector within the health assistance network, by analyzing selected information on health services and on legal rules related to supporting the private assistance subsystem. This approach recasts analyses which articulate the simultaneous examination of both the material and political instances that permanently redefine the range of activity of SUS. Considering the limitations of the empirical bases here analyzed, one questions the implied assumptions in the meanings of autonomy sustained both in the public and in the private components, departing from hypotheses on the delineation of independent tendencies toward the expansion of universal coverage and he restratification of supply and demand. PMID- 19547776 TI - Health sector reforms in comparative perspective--an unending quest ... or chasing a chimera. AB - Given the problematic nature of attempts to reform national delivery systems for health care, the paper reviews the context and logic of reforms in the health sector. After a background on approaches to building capacity within three generations of public sector reforms, it examines the reforms applied to financing, organizing and delivering health services and suggests strategies for reform that take capacity into account. PMID- 19547777 TI - UHS development and hospital services rationing. AB - We analyze Brazilian health system in comparative perspective. Middle income beneficiaries migration to pre-paid private insurance makes Brazilian case similar to United States. Public hospital services delivery shows an important retrenchment enhanced by demographic growth and new expectations due to constitutional definitions. Retrenchment is selective and concentrates on obstetric and clinic services and private for-profit services. To ensure equal access it is necessary to improve public spending; diminish out-of-pocket spending; develop organizational reforms; improve government capacity. PMID- 19547778 TI - Family health: limits and possibilities for an integral primary care approach to health care in Brazil. AB - The article analyzes the implementation of the Family Health Strategy (FH) and discusses its potential to guide the organization of the Unified Health System in Brazil, based on the integration of FH to the health care network and intersectorial action, crucial aspects of a comprehensive primary health care. Four case studies were carried out in cities with high FH coverage (Aracaju, Belo Horizonte, Florianopolis e Vitoria), using as sources: semi-structured interviews with managers and surveys with health care professionals and registered families. The integration analysis highlighted the position of FH Strategy in the health services network, the integration mechanisms and the availability of information for continuity of care. Intersectoriality was researched in relation to the fields of action, scope, sectors involved, presence of forums, and team initiatives. The results point to advances in the integration of FH to the health care network, strengthening basic services as services that are regularly sought and used as a preferential first contact services, although there are still problems in the access to specialized care. The intersectorial initiatives were broader when defined as integrated municipal government policy for the construction of interfaces and cooperation between the diverse sectors. PMID- 19547779 TI - Municipal Health Councils of Brazil: a debate on the democratization of health in the twenty years of the UHS. AB - Over 17 years, Health Councils were created in the 5,564 Brazilian towns, recruiting about 72,000 councilors. Authors affirm that the institutionalization of the Council is important for the democratization of municipal health policy, as it increases the actors who are participating in its decision-making process. However, they state that this is not enough to make this process fully democratized. This setting is investigated through a new census research about the functioning of the Municipal Health Councils. To understand it, we use three analytical dimensions (autonomy, organization and access) made up of 18 variable. The analysis of results shows that the MHCs have problems with autonomy and organization and good performance in access. Distribution by population size reveals that the best results are in the MHCs of towns with more than 250,000 inhabitants, and the worst are those in towns with a population below 50,000. The problems identified are reactions to the institutionalization of the MHCs. These reactions come from governors who consider the attributes and the deliberative character of the MHCs to be threatening to their interests. They occur due to their low cost, as the rules of the decision-making process do not discourage them. Here, we seek to understand reactions and rules, presenting proposals for overcoming problems. PMID- 19547780 TI - Federative coordination and decentralization: Brazilian experience in health. AB - This article deals with intergovernmental relations in health within the 20 years of implantation of the Unified Health System (SUS), in the light of the historical course of Brazilian federalism and its implications to health. Initially, a theoretical-conceptual review was carried out on the topic of federalism, social welfare and federative coordination of health, considering the international debate and the historical analysis of the Brazilian case. Following, the article analyzes the federal performance in the intergovernmental coordination of national health policy during the period of implantation of SUS, based on a research about the role of the Brazilian Ministry of Health from 1990 to 2002, which involved documental analysis and interviews with federal officers and other players in national politics. It was observed that health policies registered, in the past 20 years, changes in five relevant aspects that characterize federalism: institutional arrangements and rules for decisions in the federal government; the set of players with territorial basis; legal arrangements to define responsibilities among government levels; intergovernmental tax arrangements; informal arrangements among governments - vertically and horizontally. PMID- 19547781 TI - Decentralization of governmental action in Brazil in the 90s: challenges of the political-institutional environment. AB - This article analyzes the particularities of the decentralization of the state's action which took place in the Federative Republic of Brazil in the 1990 s. It presents the circumstances surrounding the political-institutional government environment which constructed the scenario in the implementation of decentralized management in the healthcare sector. It identifies challenges in rebuilding federative relations arising from the field of macroeconomic management, the social policies management and the attempts to rearrange the public sector and public administration in institutional and management terms. The aim is to reflect on the situation of federal, state and municipal governments when implementing decentralizing processes. The conclusion is that decentralization in the country was operated in the midst of the strengthening of the nation as well as of convergence and tension between two distinct projects to reform the state: a liberalizing transnational economic project of state modernization, and a national project, socially built, to expand and universalize rights and to redemocratize the state. The federative innovations, in addition pressures and influences of those two projects, determined the intergovernmental relations in reorganizing the management of public policies. PMID- 19547782 TI - State responsibility and right to health in Brazil: a balance of the Branches' actions. AB - The 1988 Federal Constitution set forth a new political-institutional moment in Brazil reasserting the Democratic State and defining a broad social protection policy including health as a social citizenship right. Since its promulgation, a great number of laws, ministerial decrees and administrative actions have attempted to make feasible the political project outlined in the Constitution. On the other hand, in the same period, the number of legal orders regarding health related demands has increased. Such a movement has revealed inconsistencies and contradictions in the legal and normative scope of SUS (Unified Health System), as well as problems not calculated by health policies, questioning the Executive Branch's actions and creating a new demand for legislation. This article discusses the role of the State in health as of 1990, considering the action of the Branches. The perspectives on the right to health in the construction of a democratic State oriented to social wellbeing, facing the challenges related to coordination mechanisms and balance among Branches in the health issue, are discussed. PMID- 19547783 TI - The financing of SUS in a scenario of financialization. AB - This article rebuilds the process of the institutionalization process of the financing of Unified Health System (SUS), impaired, initially, by the macroeconomic policy conditions, developed during the decades of 1990 and 2000, and, ultimately, by the effects caused by the present phase of capitalism, concerning financial capital supremacy. It also identifies, within the political and economic framework, conflicts existing with the economic area of the federal government, highlighting the conditions imposed to financing and the concept of health, being universal and an essential component of Social Security. PMID- 19547784 TI - Construction of the food and nutrition security policy in Brazil: strategies and challenges in the promotion of intersectorality at the federal government level. AB - This article analyzes institutional strategies of the Brazilian federal government that aim at promoting intersectorality in the field of Food and Nutrition Security (FNS), based on bibliographic review and document analysis. It is assumed that, although formal institutionality in this government level is not enough to promote intersectorality, it is important in process induction. It follows that the combination of different institutional mechanisms favors intersectorality, such as: the existence and location of councils integrated by government sectors and civil society in the presidency; political support by the presidency and inclusion of the issue as being strategic in the governmental agenda; assembly of institutional spaces that articulate the highest government spheres and that integrate technical levels; programs that integrate food production, commercialization, and consumption. Challenges concern interrelation with economic policy and the construction of budget agreed among sectors, integrated to policy management and monitoring. PMID- 19547785 TI - Evaluation of initiatives and intersectorial programs in health: challenges and learning. AB - The objective of his article is to contribute to the evaluation of the initiatives aimed at the creation of intersectorial arrangements in the scope of public health policies and social protection. The focus is on the Integrated and Sustainable Local Development of Manguinhos (DLIS-Manguinhos) - a territory and community based proposal associated to bottom up cooperation and negotiation strategies - and the experience of establishing the conditioned cash transference federal program called Bolsa Familia (Family Grant Program - PBF). In this perspective, the aspects related to social mobilization networks, profile of the players involved, types of incentive and levels of institutional integration are treated as crucial elements in the analysis of the programs and initiatives that articulate intersectorial proposals. It is concluded that the interface and dialog among research, evaluation and follow up of decision processes constitute the central axes for better social and institutional learning in the area. PMID- 19547786 TI - Medication advertising in Brazil. Can it be regulated? AB - The regulation of medication advertising in Brazil has four weak points. Inspection and punishment of irregularities is carried out a posteriori to the infraction being committed (when the population has already been exposed to a sanitary risk). The fines charged by the Brazilian Sanitary Surveillance Agency (Anvisa) have a derisory value compared to investments in advertising. There is no mechanism that prevents fines from being transferred to prices. The phrase 'If symptoms persist, consult your doctor', rather than warning about the risks of self-medication, encourages using at least the first medication without a prescription, advising a visit to the doctor only if symptoms persist. Anvisa data and academic studies reveal that 90% to 100% of advertising shown in the media contains irregularities. Thus, the Anvisa Collegiate Board of Directors Resolution 102/2000, which seeks to regulate the sector, makes up a system that benefits the infractor and keeps the population at risk. This work analyses alternative regulation, looking at advertising's previous compliance statute through the surveillance system; it studies international statutes and proposes an alteration in the structure of the current model, inserting the logic of sanitary risk prevention. PMID- 19547787 TI - Digital inclusion and health counselors: a policy for the reduction of social inequalities in Brazil. AB - Inequalities in health conditions remain even twenty years after the implementation of Unified Health System (SUS). This condition burdens social movements exerting social control on the health care area with a continuous fight. In this struggle, the accumulation of political power is related to an increase in the capacity to acquire knowledge and information. This study aims at fathoming the inequality surrounding the digital inclusion of Health Counselors (HC) of different regions within the country. We have adopted the qualitative survey method, which employs the Focal Groups technique, with HC representing managers, services providers, workers and users, all from national, state and municipal levels. Four aspects were examined, comprising reading and writing habits; Internet utilization; the use of health indicators; and the role of information in the Council-State-Society relation. Results have evidenced the need to broaden the foundations of digital inclusion initiatives in the health care area, and to overcome the cross-sector challenge of linking them to politics and education. By using benchmarks of educational philosophy, we were able to outline a theoretical-analytical matrix as a contribution to understanding the complexity involved in fostering digital inclusion in the health care area. PMID- 19547788 TI - Economic behaviour as leitmotiv in health policy in Germany and its consequences for prevention and health promotion. AB - Since the mid 1970s the health policy in Germany has focused on cost containment precisely reduction of employer's contribution for statutory health insurance (SHI). However political parties and the government have also changed their strategies to achieve this aim. The grown structures of SHI concerned with financing, provision and management remained unaltered till the early 90 s. The 90 s are marked by a broad transformation of the health sector. Regulatory instruments have been developed to create financial incentives for all stakeholders (sickness funds, health care providers, insured/patients) to reduce expenditure in health sector. The transformation process is marked by normative principle of taking stakeholders as economic subjects acting rationally. The article aims at evaluating the desired/undesired impacts of this paradigm shift, where solidarity is surpassed by competition, co-payment and privatisation. It shows where this paradigm change can be observed and which instruments have been used for its realisation. It is also given an analysis on the reaction of important stakeholders to this change. Finally, it is demonstrated ,by using examples of prevention and health promotion that the only focus on economic models doesn t allow health policy to adequately meet the challenges set upon it. PMID- 19547789 TI - Interview with Jairnilson Silva Paim: 'Taking stock of 20 years of the Unified Health System (UFS)', by Jeni Vaitsman, Marcelo Rasga Moreira e Nilson do Rosario Costa. PMID- 19547790 TI - [What do the users think and say about the Brazilian Health System (SUS)? An analysis of meanings based on the users' rights booklet]. AB - This study is the result of an expanded research-action project aimed at presenting and discussing the main thematic units that emerged from discussions conducted with a group of users about booklet regarding users' rights from the SUS (Brazilian Health System). Between May and July 2007, weekly meetings were held, recorded and followed by qualitative research data analysis until reaching the thematic units. The results show that granting access to the public health system is not enough. We must create political awareness so that users do not become mere recipients or interlocutors of government proposals. We must ensure that all social actors engage themselves in the fight for the transformation of society by making use of their rights and fulfilling their duty. PMID- 19547792 TI - [Costs with personnel and productivity analysis of family health program teams in Fortaleza, Ceara]. AB - The research verified the productivity and the operational personnel costs of eight teams of the Family Health Program (PSF) of two Basic Units of Family Health (UBASF) in Fortaleza, Ceara, through the methodology of costs by absorption, in 2004. The largest expenses were with personnel (75%), mainly with the PSF teams, and medicines (18%). The federal allocations received in September, 2004, by each PSF team, were R$ 9,543.33. The total cost by team was R$ 15,719.00. Some professionals of various PSF teams showed productivity above the set objectives, but the average productivity by doctor and nurse was below fifty per cent of objectives, with high idle time making the work onerous. The unity cost of home visits by doctor and prenatal attendance by nurses were checked, and, would be less expensive if the idle time were reduced. Various alternatives of cost reduction were observed, such as scale savings of some resources and services and re-negotiation of contracts with suppliers and cooperatives. The data obtained will contribute to more accurate planning for the installation and maintenance of PSF teams as well as alternatives of cost reductions, higher productivity, and better quality. PMID- 19547791 TI - [The needed Brazilian Health System and the possible Brazilian Health System: management strategies. An experience-based reflection]. AB - The present text presents a reflection about the author's experience as head of a Health Department of a big city during two and a half years. It presents a systematization of the strategic projects, the political and technical activities and the managerial routine, in which he was involved. It identifies three levels (macro, meso and micro) and four dimensions of management (social-political, institutional, technical-sanitary and administrative in the strict sense). In each dimension, on the three levels, it discusses management strategies designed to contribute to the construction of a universal and equitable Brazilian Health System (SUS). Although it may be premature to evaluate the degree of implantation and the effects of the proposed strategies, their analysis and discussion can be useful for being strongly based on empirical elements. The paper concludes that, even though the consolidation of the SUS is a political struggle that surpasses the management arena, managers are important agents who need to know how to develop strategies able to foster the principles of universality and equity. PMID- 19547793 TI - [Implementing health surveillance at the primary care level]. AB - This paper discusses the possibilities of re-orienting work processes at the primary care level in the light of the concepts and pre-suppositions of the health surveillance system. In addition, it presents some key concepts that could help putting into operation a health surveillance system at the local level. One of these concepts is the idea of the territory as a privileged space of primary care, helping to define and identify health needs. The study further emphasizes the heuristic value of integrating knowledge and practices in the various fields of health care so as to ensure a broader vision of problems and comprehensive health care. Finally, it analyzes the contributions from epidemiological, environmental, and health surveillance for consolidating health surveillance into a system not only limited to these three areas of action. Integrated actions of epidemiological, sanitary, and environmental surveillance can favor risk management and allow for innovative and more effective answers to the demands emerging from the health area. In addition, the local teams can acquire practical experience in internal and inter-sectorial actions which, though their importance is recognized in theory, were rarely put into practice. PMID- 19547794 TI - [Adolescents: knowledge about sexuality before and after participating in prevention workshops]. AB - Teenagers go through biological/psychosocial changes including experiencing his/her sexuality. Adolescent sexuality is often shown in unsafe sexual practices; lack of information, taboos or even the fear of accepting one s sexuality can turn it into a problem. This study aims at analyzing the knowledge of adolescents on sexuality, contraceptive methods, pregnancy, STD/AIDS before and after prevention workshops. 117 8th grade students of a public school in Londrina, Parana, ages 14 to 16, comprised the sample. A pre- and post-test questionnaire was used and data analysis was based on the chi-square and Fisher's exact tests. Boys began their sexual activity earlier than girls. In the pre test, 28.2% of the adolescents were informed about the girl's fertile period; after the workshops, this rate surpassed 55.8%. In the pre-test, AIDS was the most frequently mentioned STD; in the post-test other STDs were brought up (41.1%). The most well-known contraceptive methods were condoms and the pill. No statistically significant difference was found between the answers about risk behavior in relation to STD/AIDS transmission. Thus, it is necessary to reflect about and discuss ways of approaching the issue in schools in order to provide the adolescents with the knowledge they need to live their sexuality more safely. PMID- 19547795 TI - [Juvenile diabetes: the family's experience with diabetic adolescents and pre adolescents]. AB - This study aimed at investigating the experience of taking care of adolescents and pre-adolescents with type 1 diabetes from the standpoint of their family members. A phenomenological approach, i.e. a type of qualitative analysis aimed at understanding the meaning of life experiences, was used. Ten parents (9 mothers and 1 father) were interviewed at a teaching hospital in the interior of Sao Paulo state. The experience of taking care of a child with diabetes emerged from the converging points reported under 3 main themes: the universe of the disease; personal relationships; reflections on the experience. The participants described their difficulties and the strategies they use for keeping the family together besides providing support to the children. They believe they have to accept and face the challenges, besides motivating their children in order to ensure their safety and quality of life. The data obtained show the need for professional support as well as for a space for discussing themes of interest, both for the children with diabetes and for their families. PMID- 19547796 TI - [The historical evolution of dengue prevention and control programs in Brazil]. AB - An epidemiological analysis of the forms and distribution of Dengue fever in Brazil and worldwide was carried out. The National Program of Dengue Control (NPDC) was evaluated based on the data available at 'Diagdengue' an official computerized information system allowing to follow-up the implantation and impact of the Program. A factorial analysis was performed by means of a set of indicators contributing to variability. The indicator scores were added and the cities demanding for prior attention of the NPCD were classified for the construction of an index, which is presented three-monthly for every state, with intervals distributed between 0 and 9. The implantation of the NCPD is irregular. Cities with bad implantation are predominating in the north and northeast of the country. In the second analysis, socio-economic variables of the selected cities were added. For the association tests these cities were classified into two groups: 'Bad' and 'Good/Very Good', based on the association of the index of building infestation informed in the FAD (Yellow Fever and Dengue) database and the constructed index. Associations were verified based on variance analysis, trend test and trend estimate. It was observed that cities with low rates of illiteracy, efficient garbage collection and high ratio of sanitary installations are correlated with a good classification of the city according to Diagdengue. PMID- 19547797 TI - [Evaluation of trait and state anxiety in first year students]. AB - The purpose of this study is to evaluate trait anxiety and state anxiety in first year students from the Federal University of Rio Grande do Norte, verifying differences between three great areas of knowledge - Biomedical, Humanistic and Technological. The entrance to a University can be perceived as a threatening situation that seems to suffer influence from the different characteristics of each of these areas. 158 students, 71 females and 87 males aged 20.04 +/- 3,37 years, answered an Identification Form and the State and Trait of Anxiety Inventory (STAI). The means of trait anxiety and state anxiety observed in these students were as expected for this population although the Biomedical area is perceived as the most anxiogenic for having a dense curriculum with intense academic demands, what seems to increase the scores of the students of this area, especially of the men. PMID- 19547799 TI - Relationship between health providers and patients in Mexico City. AB - OBJECTIVE: To assess quality of the relationship between inpatients and health providers. METHODS: A qualitative study was carried out in a tertiary care hospital in the Mexico City, Mexico, between February and July 2005. In-depth interviews were conducted with 40 inpatients. The following categories of analysis were used to explore the respondent's perception of care: attitudes and actions of medical and nursing providers, effective communication, level of knowledge of patients and their family on the diagnosis, clinical treatment, and evolution. RESULTS: Overall the level of satisfaction with health care was high. Inpatients perceived everyday (non-clinical) interactions with medical and nursing providers as inadequate due to lack of confidence to request information on their condition. In addition, this perception was reinforced by excessive use by providers of technical terminology. CONCLUSIONS: The routine relationship between health providers and their patients is perceived as inadequate showing that clinical effectiveness does not mean high quality of care. There is a need to bring together technical-scientific efficiency and patients' needs and expectations of non-clinical interactions. PMID- 19547800 TI - Ministry of Health's spending on drugs: program trends from 2002 to 2007. AB - OBJECTIVE: To analyze the evolution of the Ministry of Health's spending on drugs. METHODS: The Ministry of Health's total (added) spending on drugs and its (not added) programs' were analyzed between 2002 and 2007. Actions that financed drug acquisition were obtained from the Siga Brasil system and classified according to pharmaceutical service programs. Settled values were identified for each program. For 2006 and 2007, antiretroviral drug acquisitions were analyzed. As regards drugs from the Programa de Dispensacao em Carater Excepcional (Exceptional Circumstance Drug Distribution Program), budget action data were compared to those available in the Sistema Unico de Saude (Unified Health System). Values obtained were deflated by applying the Indice de Precos ao Consumidor Amplo (Broad Consumer Price Index). Exploratory data analysis was subsequently performed. RESULTS: Spending in 2007 was 3.2 times higher than 2002 and drug spending participation in total spending rose from 5.4% in 2002 to 10.7% in 2007. Primary care drug spending increased by 75%, and strategic program spending by 124%. In the case of antiretroviral drugs, the increase was about 6%, but with an increase by 77% from 2005 to 2006, followed by a reduction by 29% from 2006 to 2007. The most significant increase in spending was observed with exceptional circumstance distribution drugs, 252% between 2003 and 2007. CONCLUSIONS: There was significant increase in drug spending between 2002 and 2007, with greater participation of antiretroviral and exceptional circumstance distribution drugs, which are comprised by a large number of patent-protected pharmaceuticals. PMID- 19547801 TI - Health service use in a population covered by the Estrategia de Saude da Familia (Family Health Strategy). AB - OBJECTIVE: To analyze health service use patterns in communities covered by the Estrategia de Saude da Familia (Family Health Strategy). METHODS: Population based cross-sectional study with a sample of 2,988 individuals, of all ages, living in areas covered by the Estrategia de Saude da Familia, in the city of Porto Alegre, Southern Brazil, between July and September 2003. Pre-coded questionnaires about demographic, socioeconomic and health information were applied to all residents of selected households. Prevalence ratios and 95% confidence intervals were calculated and chi-square tests were applied in the analyses. Poisson regression was used in the multivariable analysis to control for potential confounding factors. RESULTS: Females, persons aged 60 years or older, white, with better socioeconomic level, without health care plan coverage and with poor health self-perception were more likely to use the local family health unit. Regarding users of other health services, a similar pattern for sex, age and health self-perception was observed. However, greater use was found among people with better socioeconomic level and with health care plan coverage. CONCLUSIONS: Use of a local family health unit was greater among people from a lower socioeconomic level and without health care plan coverage, revealing the poorer individuals to be the priority of governmental actions. Model changes made to health care with the implementation of the Estrategia Saude da Familia tend to progressively improve the health conditions of the poorer population, with consequent reduction of health inequalities. PMID- 19547802 TI - Maternal supplementation with retinyl palmitate during immediate postpartum period: potential consumption by infants. AB - OBJECTIVE: To assess the effect of maternal supplementation with a single dose of retinyl palmitate during the postpartum period, in order to provide vitamin A for the infant. METHODS: A clinical trial was conducted in Natal (Northern Brazil), between March and December 2007, on 85 women distributed randomly into two groups. The postpartum supplements of retinyl palmitate consisted of a single dose of 200,000 IU (experimental group) and zero IU (control group). The retinol levels in milk were quantified using high performance liquid chromatography. Based on the retinol concentrations obtained in breast milk and through simulations, vitamin A consumption among infants 24 hours and 30 days postpartum was calculated. RESULTS: The daily provision of retinol to newborns through colostrum, 24 hours postpartum, was 1.63 micromol for the controls and 2.9 micromol for the experimental group, taking adequate intake to be 1.40 micromol/day and the milk volume consumed to be 500 ml/day. Thirty days postpartum, these values were 0.64 micromol/day (controls) and 0.89 micromol/day (experimental group), corresponding to a 39% increase in retinol concentration in the experimental group, in relation to the control group, or 64% of the recommendation for infants aged zero to six months. CONCLUSIONS: Maternal supplementation with 200,000 IU of retinyl palmitate during the immediate post partum period, and promotion of breastfeeding practices, are efficient for increasing the nutritional status of vitamin A for the mother-child pair. PMID- 19547803 TI - Vitamin A supplementation during puerperium: systematic review. AB - A systematic review on studies evaluating the effect of applying megadoses of vitamin A on the retinol concentrations in maternal milk and blood, as a short term measure for preventing hypovitaminosis A, was conducted. Based on the strategy of the Brazilian Cochrane Center for randomized trials, 115 published papers were identified in PubMed. From these, through a set of inclusion/exclusion criteria, 14 articles published between 1993 and 2007 were selected. The effects of interventions with three posological regimens (200,000, 300,000 and 400,000 IU) of vitamin A were analyzed. Out of 11 experiments conducted on maternal milk, nine presented elevation of the retinol levels in comparison with the control group; out of nine that evaluated maternal blood, four showed elevation at varying times after applying megadoses of vitamin A. It was concluded that the results from administration of vitamin A at high doses were positive in 82% of the trials on maternal milk, but less notable in relation to maternal blood. No significant differences regarding the posological regimens applied were observed. PMID- 19547804 TI - Clinical characteristics in subcortical ischemic white matter disease. AB - BACKGROUND: Vascular white matter lesions (WML) represent one of the main neuroimage findings in individuals older than 65 years and its clinical significance is still partially understood. OBJECTIVE: To describe and analyze the clinical profile of a high severity sample with WML focusing on the frontal executive control. METHOD: Outpatients (n=20) with high severity WML evaluated with magnetic resonance imaging were selected using the Fazekas scale. RESULTS: Most patients (n=17; 85%) presented an altered Trail Making Test ratio (section B/section A); on verbal fluency, 15 individuals (75%) performed below the cutoff score. Apathy (5.9 +/- 4.65) and depression (3.05+/-3.67) were frequent as assessed by the Neuropsychiatric Inventory. The impairment in functional activities strongly correlated with apathy (r=0.814, p<0.001) and verbal fluency (r=0.744, p<0.001). CONCLUSION: Executive dysfunction, apathy, and ratio depression were the main characteristics found. Extension of WML may have distinct impact on the clinical picture, but further studies with methodological adjustments are necessary to provide more definitive conclusions. PMID- 19547805 TI - Binswanger's disease and quantitative fractional anisotropy. AB - OBJECTIVE: To study the integrity of the white matter in Binswanger's disease (BD) patients with quantitative fractional anisotropy (DTI-FA). METHOD: Controls (12) and patients with BD (12) were included. Scans performed with MR (GE Signa Horizon/1.5T). Fazekas's score=6 with white matter hyperintensities extension >75% assessed on FLAIR scans. Standard parameters for DTI-FA were used. ROIs placed in symmetrical regions on two axial planes, data pooled in anterior (frontal) and posterior (temporo-parieto-occipital) regions. Analysis with Functool. Statistics for anterior and posterior regions comparison. RESULTS: DTI FA showed reduction of anisotropy, reflecting axonal damage and demyelination of fibers, more prominent in anterior in relation to posterior region, in BD patients in comparison to controls. CONCLUSION: Loss of integrity of fiber tracts reflects interruption of neural networks that subserve cognitive, behavioral, and motor integration. The more severely affected frontal region is related to executive dysfunction, a characteristic feature of Binswanger's disease. PMID- 19547806 TI - Combined instruments for the screening of dementia in older people with low education. AB - OBJECTIVE: To determine which combination of cognitive tests and informant reports can improve the diagnostic accuracy of dementia screening in low educated older people. METHOD: Patients with mild to moderate dementia (n=34) according to ICD-10 and DSM-III-R criteria and 59 older controls were assessed with the Mini Mental State Examination (MMSE) and the Fuld Object Memory Evaluation (FOME). Informants were assessed using the Informant Questionnaire on Cognitive Decline in the Elderly and the Bayer-Activities of Daily Living Scale. RESULTS: The 4 instruments combined with the mixed rule correctly classified 100% and the logistic regression (weighted sum) classified 95.7% of subjects. The weighted sum had a significantly larger ROC area compared to MMSE (p=0.008) and FOME (p=0.023). The specificity of the tested combinations was superior to the MMSE alone (p=0.002). CONCLUSIONS: Cognitive tests combined with informant reports can improve the screening of mild to moderate dementia in low educated older people. PMID- 19547807 TI - Association between the DRD2-141C Insertion/Deletion polymorphism and schizophrenia. AB - Epidemiological studies have demonstrated that the genetic component is an important risk factor for the development of schizophrenia. The genes that codify the different compounds of the dopaminergic system have created interest for molecular investigations in patients with schizophrenia because the antipsychotic drugs, especially those of first generation, act on this cerebral system. Thus the aim of the present study was to investigate the possible association between the -141 Ins/Del (rs1799732) polymorphism of the dopamine receptor type 2 (DRD2) and schizophrenia. The distribution of the alleles and genotypes of the studied polymorphism was investigated in a sample of 229 patients and 733 controls. There were statistical differences in the allelic (chi2=9.78; p=0.001) and genotypic genotypic (chi2=12.74; p=0.001) distributions between patients and controls. Thus the -141C Ins/Del polymorphism of the DRD2 gene (allele Ins) was associated to the SCZ phenotype in the investigated sample. PMID- 19547808 TI - Effectiveness of clozapine, haloperidol and chlorpromazine in schizophrenia during a five-year period. AB - OBJECTIVE: The aim of our study was to evaluate the effects of low doses of clozapine in flexible regime in comparison with haloperidol and chlorpromazine in long term. METHOD: The naturalistic study was prospective, active-controlled with 325 adult outpatients of both genders (140 females), with mean year age of 34.8 (range 21-57), suffering from chronic schizophrenia. The first onset of illness was at the mean of 27.9 years (range 17-38), and subjects had the mean year age of 4.1+/-0.5 previous relapses. The patients were allocated to receive haloperidol (105 subjects, dose range 2-15 mg), chlorpromazine (n=105, 100-400 mg) or clozapine (n=115, 75-600 mg). The scores of psychometric instruments (GWB, PANSS, CGI) were regularly assessed during 5 year period. RESULTS: The sixty-six responders were included in per-protocol analysis: 12, 10 and 16 with positive and 7, 6 and 15 with negative schizophrenic syndrome in haloperidol, chlorpromazine and clozapine group, respectively. The statistically significant differences in all psychometric scores was found, for both schizophrenic syndromes, favoring clozapine. The distribution of eighteen different types of adverse events, which we noted, were significantly different among treatment groups ( chi2=315.7, df=34, p<0.001). Clozapine was safer and had fewer adverse effects (average of 0.9 adverse events per patient) than haloperidol (2.7) and chlorpromazine (3.2). CONCLUSIONS: Clozapine, in low doses of flexible regime, in long term (five years) showed better effectiveness in chronic schizophrenics with positive and negative symptoms than typical antipsychotics. PMID- 19547809 TI - Depressive symptoms and perception of quality of life in Parkinson's disease. AB - BACKGROUND: Depression has been proposed as a major contributor to poor quality of life (QoL) in Parkinson's disease (PD). OBJECTIVE: To evaluate the relationship between depressive symptoms and QoL in subjects with PD. METHOD: Beck Depression Inventary (BDI) was used to evaluate depressive symptoms and Parkinson's Disease Quality of Life Questionnaire (PDQ-39) to assess the perception of the QoL. RESULTS: Thirty seven patients (19 male/ 18 female) with a typical onset PD and mean disease duration of 7.7 years were studied. Higher scores on BDI correlated with poorer perception of the QoL. This association occurred at the expense of the following PDQ39 domains: mobility, activities of daily living, social support, cognition and emotional well-being dimensions. PD severity also correlated with QoL. CONCLUSION: Our study corroborates the assumption that depressive symptoms contributed significantly to QoL in PD. PMID- 19547810 TI - Is there something special about cardiovascular abnormalities and sudden unexpected death in epilepsy among patients with chronic renal insufficiency in regular hemodialysis program? AB - Of the many risk factors suggested for sudden unexpected death in epilepsy (SUDEP), higher frequency of seizures is a very consistent issue. Following this reasoning, it has been established that hemodialysis-associated seizure is a complication of dialysis procedure. Based on these facts, this study investigated a possible association between cardiovascular abnormalities and SUDEP among patients with chronic renal insufficiency in regular hemodialysis program. For that, a retrospective medical history of 209 patients was reviewed to investigate the occurrence of convulsive seizures and EKG abnormalities during dialytic program. Three patients presented generalized tonic-clonic seizures, one had partial seizure with secondary generalization, and one presented unclassified seizure. Any EKG abnormalities and SUDEP event were found in all patients evaluated. In conclusion, the present findings demonstrated uncommon the occurrence of seizures and also SUDEP. Probably, the main justification to not allow us to demonstrated a direct relation between SUDEP and cardiovascular diseases in hemodialysis are the reduced number of cases examined. PMID- 19547811 TI - Effects of a cognitive modulator in the theta and alpha asymmetry during a typewriting task: a sensorimotor integration perspective. AB - This study aimed to elucidate cortical mechanisms and to identify the areas where occur such mechanisms due to interaction between bromazepam and motor learning. The sample was composed of 45 healthy subjects randomly distributed in 3 groups: placebo (n=15), bromazepam 3 mg (n=15) or bromazepam 6 mg (n=15). To perform the experimental task, subjects sat comfortably at a distance of approximately 20 cm from the typewriter. The typewriter keyboard was covered with a wooden box to avoid visual information about the hands' position. The typewriting task was performed concomitantly with EEG recording. ANOVA two-way results indicated a decreased asymmetry in sensorimotor areas in the experimental groups. Our interpretation is that moderate doses of bromazepam may improve performance on tasks with predictable elements to promote stability of psychomotor functions, but may also impair performance on tasks executed in unpredictable environments. PMID- 19547812 TI - Vestibular rehabilitation: clinical benefits to patients with Parkinson's disease. AB - OBJECTIVE: To evaluate the effectiveness of the vestibular rehabilitation (VR) exercises by means of an assessment before and after the application of the Brazilian version of the Dizziness Handicap Inventory (DHI) questionnaire. METHOD: Twelve patients were studied, the following procedures were carried out: anamnesis, otorhinolaryngological and vestibular evaluation, and the application of the DHI before and after the VR. RESULTS: Clinically resting tremors and subjective postural instability were the motor complaints most frequently associated with complaints of vertigo in 12 cases (100%); in the vestibular exam, all the patients presented abnormalities, frequently from the uni and bilateral peripheral vestibular deficiency syndromes in 10 cases (83.3%); there was significant improvement in the physical, functional and emotional aspects of the DHI after the completion of the VR. CONCLUSION: The VR following the Cawthorne and Cooksey protocol were shown to be useful in managing subjective complaints of several aspects evaluated in this protocol. PMID- 19547813 TI - Performance of normal adults on Rey Auditory Learning Test: a pilot study. AB - The present study aimed to assess the performance of healthy Brazilian adults on the Rey Auditory Verbal Learning Test (RAVLT), a test devised for assessing memory, and to investigate the influence of the variables age, sex and education on the performance obtained, and finally to suggest scores which may be adopted for assessing memory with this instrument. The performance of 130 individuals, subdivided into groups according to age and education, was assessed. Overall performance decreased with age. Schooling presented a strong and positive relationship with scores on all subitems analyzed except learning, for which no influence was found. Mean scores of subitems analyzed did not differ significantly between men and women, except for the delayed recall subitem. This manuscript describes RAVLT scores according to age and education. In summary, this is a pilot study that presents a profile of Brazilian adults on A1, A7, recognition and LOT subitem. PMID- 19547814 TI - Potential risk factors for multiple sclerosis in Rio de Janeiro: a case-control study. AB - PURPOSE: To evaluate potential risk factors for the development of multiple sclerosis in Brazilian patients. METHOD: A case control study was carried out in 81 patients enrolled at the Department of Neurology of the Hospital da Lagoa in Rio de Janeiro, and 81 paired controls. A standardized questionnaire on demographic, social and cultural variables, and medical and family history was used. Statistical analysis was performed using descriptive statistics and conditional logistic regression models with the SPSS for Windows software program. RESULTS: Having standard vaccinations (vaccinations specified by the Brazilian government) (OR=16.2; 95% CI=2.3-115.2), smoking (OR=7.6; 95% CI=2.1 28.2), being single (OR=4.7; 95% CI=1.4-15.6) and eating animal brain (OR=3.4; 95% CI=1.2-9.8) increased the risk of developing MS. CONCLUSIONS: RESULTS of this study may contribute towards better awareness of the epidemiological characteristics of Brazilian patients with multiple sclerosis. PMID- 19547815 TI - Predictive factors for neuromotor abnormalities at the corrected age of 12 months in very low birth weight premature infants. AB - BACKGROUND: The increase in survival of premature newborns has sparked growing interest in the prediction of their long-term neurodevelopment. OBJECTIVE: To estimate the incidence of neuromotor abnormalities at the corrected age of 12 months and to identify the predictive factors associated with altered neuromotor development in very low birth weight premature infants. METHOD: Cohort study. The sample included 100 premature infants. The outcome was neuromotor development at 12 months classified by Bayley Scale (PDI) and neurological assessment (tonus, reflexes, posture). A multivariate logistic regression model was constructed. Neonatal variables and neuromotor abnormalities up to 6 months of corrected age were selected by bivariate analysis. RESULTS: Mean birth weight was 1126g (SD: 240). Abnormal neuromotor development was presented in 60 children at 12 months corrected age. CONCLUSION: According to the model, patients with a diagnosis including bronchopulmonary dysplasia, hypertonia of lower extremities, truncal hypotonia showed a 94.0% probability of neuromotor involvement at 12 months. PMID- 19547816 TI - Language and cognition in very low birth weight preterm infants with PELCDO application. AB - The performance of very low birth weight preterm infants, in terms of cognition and expressive language, was analyzed and compared with that of term infants with the Protocol for Expressive Language and Cognition Development Observation (PELCDO). The study involved 12 very low birth weight preterm infants and 20 term infants, all of whom were evaluated monthly. Sessions were videotaped, and data were analyzed according to this specific protocol. Our results suggest that cognition and expressive language develop significantly later in very low birth weight preterm infants than in the term. We found positive correlations for cognitive and expressive language development, the delay becoming more evident after 6 months of age, persisting through the sensorimotor period, and continuing into the beginning of preoperational period, indicating the importance of follow up evaluation, defining the true needs of such infants and identifying the ideal moment for speech-language intervention. PMID- 19547817 TI - Application of magnetic resonance spectroscopy in the differentiation of high grade brain neoplasm and inflammatory brain lesions. AB - This study aims at evaluating the application of magnetic resonance spectroscopy (MRS) in the differential diagnosis of brain tumors and inflammatory brain lesions. The examinations of 81 individuals, who performed brain MRS and were retrospectively analyzed. The patients with ages between 10 and 80 years old, were divided into two groups. Group A consisted of 42 individuals with diagnoses of cerebral toxoplasmosis and Group B was formed of 39 individuals with diagnosis of glial neoplasms. On analyzing the ROC curve, the discriminatory boundary for the Cho/Cr ratio between inflammatory lesions and tumors was 1.97 and for the NAA/Cr ratio it was 1.12. RMS is an important method useful in the distinction of inflammatory brain lesions and high-degree tumors when the Cho/Cr ratio is greater than 1.97 and the NAA/Cr ratio is less than 1.12. And so this method is important in the planning of treatment and monitoring of the therapeutic efficiency. PMID- 19547818 TI - Proton magnetic resonance spectroscopy in children with fetal alcohol spectrum disorders. AB - OBJECTIVE: To analyze the metabolic constitution of brain areas through proton magnetic resonance spectroscopy in children affected with fetal alcohol spectrum disorder compared with normal children. METHOD: The sample of this case-control study included eight boys with epidemiologic history of in utero exposure to alcohol (median age 13.6+/-3.8 years) who were diagnosed with fetal alcohol spectrum disorder, and eight controls (median age 12.1+/-3,4 years). An 8 cm(3) single voxel approach was used, with echo time 30 ms, repetition time 1500 ms, and 128 acquisitions in a 1.5T scanner, and four brain areas were analyzed: anterior cingulate, left frontal lobe, left striatum, and left cerebellar hemisphere. Peaks and ratios of metabolites N-acetylaspartate, choline, creatine, and myo-inositol were measured. RESULTS: Children with fetal alcohol spectrum disorder showed a decrease in choline/creatine ratio (p=0.020) in left striatum and an increase in myo-inositol/creatine ratio (p=0.048) in left cerebellum compared with controls. There was no statistically significant difference in all peaks and ratios from the anterior cingulate and frontal lobe between the two groups. CONCLUSION: This study found evidence that the left striatum and left cerebellum are affected by intrauterine exposure to alcohol. Additional studies with larger samples are necessary to expand our knowledge of the effects of fetal exposure to alcohol. PMID- 19547819 TI - C5 and C6 human dermatomes: a clinical, electromyographical, imaging and surgical findings. AB - There is substantial controversy in literature about human dermatomes. In this work, C5 and C6 superior limb dermatomes were studied. The method consisted of comparing clinical signs and symptoms with conduction studies, electromyographical data, neurosurgical findings, and imaging findings obtained by computerized tomography (CT) or magnetic resonance imaging (MRI), for each patient. Data analysis from superior members in 18 patients suggests that C5 is located in the lateral aspect of the shoulder and arm, and C6 in the lateral aspect of the forearm and 1st, 2nd, and 3rd fingers. To our knowledge this is the first time that C5 and C6 human dermatomes have been studied by all the following methods together: clinical, electromyographical, CT and MR imaging, and surgical findings. PMID- 19547820 TI - L4-L5-S1 human dermatomes: a clinical, electromyographical, imaging and surgical findings. AB - There is substantial controversy in literature about human dermatomes. We studied L4, L5, and S1 inferior limb dermatomes by comparing clinical signs and symptoms with conduction studies, electromyographical data, neurosurgical findings, and imaging data from computerized tomography (CT) or magnetic resonance imaging (MRI). After analyzing 60 patients, we concluded that L4 is probably located in the medial aspect of the leg, L5 in the lateral aspect of the leg and foot dorsus, and S1 in the posterior aspect of the backside, tight, leg and plantar foot skin. This is the first time that these human dermatomes have been evaluated by combined analysis of clinical, electromyographical, neurosurgical, and imaging data. PMID- 19547822 TI - Early shunt complications in 46 children with hydrocephalus. AB - OBJECTIVE: To determine the causes of early shunt complications in 46 children with hydrocephalus. METHOD: A retrospective study was conducted on 46 children submitted to ventriculoperitoneal shunt surgery between February 2005 and February 2007. RESULTS: Thirteen (28%) patients presented complications, which were due to infection in 9 (69%) and to malfunction of the shunt system in 4 (31%).The mean number of surgical procedures performed on patients who presented complications was 2.8 per patient, with a total of 46 surgeries in this group. All patients with infectious complications were identified during their hospital stay. CONCLUSIONS: Infection was the most common complication. The infection rate was proportional to the length of hospital stay. All patients with hydrocephalus due to tumors or myelomeningocele presented complications. A higher incidence of infections was observed in children older than 2 years. PMID- 19547821 TI - Transitional lumbosacral vertebrae and low back pain: diagnostic pitfalls and management of Bertolotti's syndrome. AB - OBJECTIVE: Bertolotti's syndrome is a spine disorder characterized by the occurrence of a congenital lumbar transverse mega-apophysis in a transitional vertebral body that usually articulates with the sacrum or the iliac bone. It has been considered a possible cause of low back pain. METHOD: We analyzed the cases of Bertolotti's syndrome that failed clinical treatment and reviewed the literature concerning this subject. RESULTS: Five patients in our series had severe low back pain due to the neo-articulation and two of them were successfully submitted to surgical resection of the transverse mega-apophysis. Taking into account the clinical and surgical experience acquired with these cases, we propose a diagnostic-therapeutic algorithm. CONCLUSION: There is still no consensus about the most appropriate therapy for Bertolotti's syndrome. In patients in whom the mega-apophysis itself may be the source of back pain, surgical resection may be a safe and effective procedure. PMID- 19547823 TI - Spontaneous thrombosis of internal carotid artery: a natural history of giant carotid cavernous aneurysms. AB - OBJECTIVE: To describe five cases of giant carotid cavernous aneurysms which evolved with spontaneous thrombosis of internal carotid artery (STICA), with emphasis at epidemiology, clinical presentation, natural history, related factors and neurological outcome. METHOD: There were 711 consecutives patients with 802 aneurysms with and without surgical treatment during a period of 19 years. We selected 35 patients with 40 carotid cavernous aneurysms (5%) of which 20 (50%) were giant aneurysms. Among those cases, 5 patients evolved with STICA (25%). Symptoms and findings at presentation were recorded and compared with those at outcome. RESULTS: Clinical presentation was commonly related to atherosclerotic factors such as elevated blood pressure (80%), diabetes mellitus (40%) and dislipidemy (40%). All patients presented with hemicranial headache, ophthalmparesy and retro bulbar pain, and after STICA all presented improvement of symptoms. After STICA, 4 patients had regression of deficit, 2 partial and 2 complete. Four patients had sensorial trigeminal neuropathy in V1 and V2 territories, also showing improvement of symptoms after STICA. CONCLUSION: STICA is a common outcome in giant carotid cavernous aneurysms, and is related with significant improvement of symptoms; however, it may be catastrophic for those patients without efficient collateral circulation. PMID- 19547824 TI - Surgical management of intramedullary spinal ependymomas. AB - BACKGROUND: Spinal intramedullary ependymoma is a rare disease with a wide range of clinical presentation, generally requiring surgical treatment. OBJECTIVE: Report our experience and present our surgical technique to achieve total resection and cure. METHOD: We present 12 consecutive cases of intramedullary ependymomas operated between 2000 and 2008 by the senior author (HT). The functional scale proposed by McCormick was used to evaluate the patients' neurological status. RESULTS: Age at presentation varied from 18 to 55 (average 36) years. All tumors had a benign histology. Four (33%) patients were male and eight (67%) were female. According to the site of presentation, six (50%) were localized at the cervical region (including two at the cervicomedullary junction, two at the cervico-thoracic junction and two exclusively at the cervical level), four at the thoracic level and two at the conus/ cauda equina. Dyshestesia was a common finding at the neurological exam in eight patients (67%). Total resection was achieved in all cases. Six patients showed neurological improvement postoperatively, whereas the other six remained stable. CONCLUSION: Adequate knowledge of anatomy and the correct use of microsurgical techniques allowed total resection of these tumors with minimal morbidity and maximum functional recovery. PMID- 19547825 TI - Cerebellar cryptococcoma simulating metastatic neoplasm. PMID- 19547826 TI - Concomitant dermoid cysts of conus medullaris and cauda equina. PMID- 19547827 TI - Levodopa responsive dystonia secondary to virus encephalitis. PMID- 19547828 TI - Self-aggression and congenital clubfoot: additional features to the septo-optic dysplasia complex. PMID- 19547829 TI - Brain tumor or infectious disease? PMID- 19547830 TI - Oculocerebrorenal syndrome of Lowe: magnetic resonance imaging findings in the first six years of life. PMID- 19547831 TI - Subdural haematoma in a patient with meningioma. PMID- 19547832 TI - Chronic inflammatory demyelinating polyneuropathy in an adolescent with type 1 diabetes mellitus. PMID- 19547833 TI - The influence of circadian rhythms on sudden unexpected death in epilepsy. PMID- 19547834 TI - Angioarchitecture and clinical presentation of brain arteriovenous malformations. AB - The purpose of this study was to correlate the angioarchitecture of brain arteriovenous malformations (AVM) with their clinical presentation. A total of 170 patients with AVM 78 males and 92 females, were studied. Univariate and multivariate analyses were conducted in order to test the associations between morphological features and clinical presentation. The most frequent clinical presentations at diagnosis were hemorrhage in 89 (52%) patients, headache in 79 (46%), focal neurological deficit in 54 (32%), and seizure in 52 (31%). According to the Spetzler-Martin classification, grade I was found in 15 patients, II in 49, III in 55, IV in 41, and grade V in 10 patients. AVM with small nidus size, single feeding artery and single draining vein were associated with hemorrhage. Hemorrhage was positively associated with Spetzler-Martin grade I and negatively with grade V. The association between seizure and large nidus size was positive, however negative with small nidus size. PMID- 19547835 TI - Konstantin N. Tretiakoff in Brazil: a historical perspective and discussion of his contribution to brazilian neuroscience. AB - The Hospicio de Juquery, near the city of Sao Paulo (Brazil) was founded in 1896 and after few years it was decided that the institution should have the best possible facilities to study neuropathology. In 1921, a young psychiatrist, Antonio Carlos Pacheco e Silva was sent to the Hopital de la Salpetriere (Paris) to study neuropathology. There, Pacheco e Silva (later Prof.Pacheco e Silva) befriended Konstantin N. Tretiakoff accepted an invitation to become the first Chairman of the newly created neuropathology department of the Hospicio de Juquery. During his stay in this institution, from 1922 to 1924 or early 1925, he worked very hard and produced many publications. Here we present and comment some of the papers he published in a Journal (Memorias do Hospicio de Juquery - 'Memoirs de l'Hospice de Juquery'), which had been recently created and present some information of this poorly known period of his life. PMID- 19547836 TI - Pituitary tumor apoplexy. AB - Pituitary tumor apoplexy is a medical emergency due to acute infarction or hemorrhage in the pituitary gland. In this review, the authors discuss the sellar anatomy, the pituitary gland and adenomas' vascularization and the general aspects of the syndrome such as its ethiopatogenesis, predisposing factors, clinical features, treatment and prognosis. PMID- 19547837 TI - Alzheimer's disease and implicit memory. AB - Specific neuropsychiatric disorders, such as Alzheimer's disease (AD) affect some forms of memory while leaving others relatively intact. In this review, we investigate particularities of the relationship between explicit and implicit memories in AD. It was found that implicit memory is preserved in AD, irrespective of the task used; in other words, there was not interference from explicit memory. In addition, it was verified that is possible through implicit memory compensatory strategies such as, activities of daily living (ADL) to compensate for the explicit memory deficits. In this sense, cognitive rehabilitation (CR) demonstrates reasonable results in the process of compensation of explicit memory deficits. Concluding, the decline in explicit memory suggests that both systems are functionally independent even if the other is compromised. We expect that when explicit memory system is not involved in competition with the implicit system, the final effect of learning is better, because all of the implicit memory capacity is engaged in learning and not in competition with the explicit system. PMID- 19547838 TI - Congenital muscular dystrophy. Part II: a review of pathogenesis and therapeutic perspectives. AB - The congenital muscular dystrophies (CMDs) are a group of genetically and clinically heterogeneous hereditary myopathies with preferentially autosomal recessive inheritance, that are characterized by congenital hypotonia, delayed motor development and early onset of progressive muscle weakness associated with dystrophic pattern on muscle biopsy. The clinical course is broadly variable and can comprise the involvement of the brain and eyes. From 1994, a great development in the knowledge of the molecular basis has occurred and the classification of CMDs has to be continuously up dated. In the last number of this journal, we presented the main clinical and diagnostic data concerning the different subtypes of CMD. In this second part of the review, we analyse the main reports from the literature concerning the pathogenesis and the therapeutic perspectives of the most common subtypes of CMD: MDC1A with merosin deficiency, collagen VI related CMDs (Ullrich and Bethlem), CMDs with abnormal glycosylation of alpha-dystroglycan (Fukuyama CMD, Muscle-eye-brain disease, Walker Warburg syndrome, MDC1C, MDC1D), and rigid spine syndrome, another much rare subtype of CMDs not related with the dystrophin/glycoproteins/extracellular matrix complex. PMID- 19547840 TI - Dom Pedro II daytime sleepiness. PMID- 19547841 TI - Tumour-like chagasic encephalitis in AIDS patients. PMID- 19547842 TI - Absence of MRI exams in epidemiological studies can leads to clinical overdiagnosis of Alzheimer's disease and underdiagnosis of vascular dementia. PMID- 19547843 TI - Professor Charles David Marsden. PMID- 19547844 TI - Did Darwin have syncope caused by Chagas' disease? PMID- 19547845 TI - Influenza and scientific articles. PMID- 19547846 TI - Assessment of exercise capacity in pulmonary hypertension. PMID- 19547847 TI - Accuracy of clinical examination findings in the diagnosis of COPD. AB - OBJECTIVE: Simple diagnostic methods can facilitate the diagnosis of COPD, which is a major public health problem. The objective of this study was to investigate the accuracy of clinical variables in the diagnosis of COPD. METHODS: Patients with COPD and control subjects were prospectively evaluated by two investigators regarding nine clinical variables. The likelihood ratio for the diagnosis of COPD was determined using a logistic regression model. RESULTS: The study comprised 98 patients with COPD (mean age, 62.3+/- 12.3 years; mean FEV1, 48.3 +/- 21.6%) and 102 controls. The likelihood ratios (95% CIs) for the diagnosis of COPD were as follows: 4.75 (2.29-9.82; p < 0.0001) for accessory muscle recruitment; 5.05 (2.72-9.39; p < 0.0001) for pursed-lip breathing; 2.58 (1.45-4.57; p < 0.001) for barrel chest; 3.65 (2.01-6.62; p < 0.0001) for decreased chest expansion; 7.17 (3.75-13.73; p < 0.0001) for reduced breath sounds; 2.17 (1.01-4.67; p < 0.05) for a thoracic index > or = 0.9; 2.36 (1.22-4.58; p < 0.05) for laryngeal height < or = 5.5 cm; 3.44 (1.92-6.16; p < 0.0001) for forced expiratory time > or = 4 s; and 4.78 (2.13-10.70; p < 0.0001) for lower liver edge > or = 4 cm from lower costal edge. Inter-rater reliability for those same variables was, respectively, 0.57, 0.45, 0.62, 0.32, 0.53, 0.32, 0.59, 0.52 and 0.44 (p < 0.0001 for all). CONCLUSIONS: Various clinical examination findings could be used as diagnostic tests for COPD. PMID- 19547848 TI - Association between nutritional status measurements and pulmonary function in children and adolescents with cystic fibrosis. AB - OBJECTIVE: To evaluate the association between nutritional status measurements and pulmonary function in children and adolescents with cystic fibrosis. METHODS: We evaluated the nutritional status of 48 children and adolescents (aged 6-18 years) with cystic fibrosis based on body mass index (BMI) and body composition measurements-mid-arm muscle circumference (MAMC) and triceps skinfold thickness (TST)-at a referral center in the city of Rio de Janeiro, Brazil. Pulmonary function was assessed by means of spirometry, using FEV1 to classify the severity of airway obstruction. We used Student's t-tests for comparisons between proportions and linear regression analysis for associations between continuous variables. The level of significance was set at p < 0.05. RESULTS: The evaluation of nutritional status based on BMI identified a smaller number of malnourished patients than did that based on MAMC (14 vs. 25 patients, respectively). Most of the patients presented mild pulmonary disease. Mean FEV1 was 82.5% of predicted. Pulmonary function was found to correlate significantly with BMI, MAMC and TST (p = 0.001, p = 0.001 and p = 0.03, respectively). All subjects with moderate or severe pulmonary involvement were considered malnourished based on BMI and body composition parameters. Of the 25 patients considered malnourished based on body composition (MAMC), 19 were considered well-nourished based on their BMI. CONCLUSIONS: In the present study, all nutritional status measurements correlated directly with the pulmonary function of children and adolescents with cystic fibrosis. However, body composition measurements allowed earlier detection of nutritional deficiencies. PMID- 19547849 TI - Comparison of three sets of reference equations for spirometry in children and adolescents with distinct body mass indices. AB - OBJECTIVE: To compare FEV1 and FVC, calculated using three sets of reference equations (devised by Polgar & Promadhat, Hsu et al. and Mallozi in 1971, 1979 and 1995, respectively) and to determine whether the three are similar in predicting lung function in children and adolescents with distinct body mass indices (BMIs). METHODS: The individuals were separated into four groups in accordance with the reference standards of the National Center for Health Statistics: underweight (UW), normal weight (NW), overweight (OW), and obese (OB). All were then submitted to spirometry. RESULTS: We evaluated 122 healthy children and adolescents, aged 7-14 years. The FVC values predicted for NW females and UW males through the use of the Hsu et al. equation were significantly higher than the measured values, as were the FEV1 values for UW females and males predicted via the Polgar & Promadhat and Hsu et al. equations. In NW females, the FEV1 values predicted via the Polgar & Promadhat equation were significantly higher than were the measured values. CONCLUSIONS: In individuals with distinct BMIs, the measured FVC and FEV1 values were not equivalent to those predicted via the Polgar & Promadhat and Hsu et al. equations. The same was not true for the Mallozi equations. The BMI was not a relevant factor for the predictive index of these equations; therefore, the Mallozi equations can be used without alteration for children and adolescents with distinct BMIs. PMID- 19547850 TI - Validation of a treadmill six-minute walk test protocol for the evaluation of patients with pulmonary arterial hypertension. AB - OBJECTIVE: To develop and validate a protocol for the treadmill six-minute walk test (tread6MWT) to evaluate patients with pulmonary arterial hypertension (PAH). METHODS: The study population comprised 73 patients with PAH, diagnosed by means of right heart catheterization, with or without NO inhalation. All patients performed a hallway 6MWT and three tread6MWTs based on a pre-determined incremental speed protocol and interposed by a rest period. The patients who had been submitted to hemodynamic testing using NO performed the third tread6MWT while inhaling the same dose of NO that had been used during the catheterization. RESULTS: We found that the treadmill six-minute walk distance (tread6MWD) correlated with hemodynamic data, functional class and the hallway six-minute walk distance (6MWD). In addition, the tread6MWD correlated significantly with survival, thereby confirming the correlation with disease severity. Inhalation of NO during the tread6MWT led to variations that were consistent with the hemodynamic changes induced by the same dose of inhaled NO, suggesting that the protocol developed can reflect the effect of therapeutic interventions. CONCLUSIONS: We conclude that the tread6MWD is a useful prognostic and functional marker for the routine evaluation of PAH patients. PMID- 19547851 TI - Bacterial pneumonia following bone marrow transplantation: HRCT findings. AB - OBJECTIVE: To describe HRCT findings in patients with bacterial pneumonia following bone marrow transplantation (BMT). METHODS: This was a retrospective study involving 30 patients diagnosed with bacterial pneumonia in whom HRCT of the chest was performed within 24 h after the onset of symptoms and the diagnosis was confirmed, based on a positive culture of sputum or bronchial aspirate, together with a positive pleural fluid or blood culture, within one week after symptom onset. There were 20 male patients and 10 female patients. The median age was 21 years (range, 1-41 years). The BMT had been performed for the treatment of the following: chronic myeloid leukemia, in 14 cases; severe aplastic anemia, in 6; acute myeloid leukemia, in 4; Fanconi's anemia, in 3; and acute lymphocytic leukemia, in 3. Two radiologists analyzed the HRCT scans and reached their final decisions by consensus. RESULTS: The most common HRCT findings were air-space consolidation (in 60%), small centrilobular nodules (in 50%), ground-glass opacities (in 40%), bronchial wall thickening (in 20%), large nodules (in 20%), pleural lesions (in 16.7%) and tree-in-bud opacities (in 10%). The pulmonary lesions were distributed in the central and peripheral areas in 15 patients, whereas they were exclusively peripheral in 11. Lesions were located in the lower and middle lobes of the lung in 22 and 20 patients, respectively. CONCLUSIONS: The most common HRCT findings in our patient sample were air-space consolidation, small centrilobular nodules and ground-glass opacities, most often in the central and peripheral regions of the middle and lower lung zones. PMID- 19547852 TI - The impact of smoking cessation on patient quality of life. AB - OBJECTIVE: To evaluate changes in health-related quality of life (HRQoL) after twelve months of smoking cessation. METHODS: This was a prospective study to evaluate the effectiveness of a smoking cessation program on the quality of life of 60 self-referred subjects, at a public hospital, during the period of August 2006 to December 2007. The program consisted of 2-h group sessions once a week during the first month and then every 15 days over six months, followed by monthly phone contacts for another six months. The treatment was based on behavior modification and the use of bupropion in combination with nicotinic replacement therapy. Abstinence was verified by exhaled CO measurements. Patient HRQoL was quantified using the Medical Outcomes Study 36-item Short-Form Health Survey (SF-36) questionnaire. Differences in quality of life scores between quitters and non-quitters at twelve months after the initial intervention were evaluated using analysis of covariance with baseline characteristics as covariates. RESULTS: Self-reported quality of life scores were significantly higher among the 40 quitters than among the 20 non-quitters. The following SF-36 domains were most affected: role-emotional (p = 0.008); general health (p = 0.006); vitality (p < 0.001); and mental health (p = 0.002). At twelve months after the smoking cessation intervention, the SF-36 mental component and physical component summary scores were higher among quitters than among non-quitters (p = 0.004 and p = 0.001, respectively). CONCLUSIONS: Our findings illustrate that smoking abstinence is related to better HRQoL, especially in aspects of mental health. PMID- 19547853 TI - Prevalence of and variables related to smoking among medical students at a university in the city of Passo Fundo, Brazil. AB - OBJECTIVE: To determine the prevalence of and factors associated with smoking among medical students, as well as to evaluate the profile of this group. METHODS: A total of 316 medical students (98.7% of the total) at the University of Passo Fundo, in the city of Passo Fundo, Brazil, completed a self-report questionnaire with questions on tobacco intake and on attitudes related to smoking. In accordance with the World Health Organization guidelines, the students were classified as daily smokers, occasional smokers, former smokers or nonsmokers, those in the two first categories being considered active smokers. RESULTS: We found that 16.5% of the students were active smokers (daily smokers, 5.4%; occasional smokers, 11.1%) and that 3.5% were former smokers. The mean age was 22.2 +/- 2.4 years. Factors significantly associated with the smoking habit (p < 0.05) were male gender, paternal smoking, regular alcohol consumption and use of antidepressants or anxiolytics. For the majority (69.2%) of the smokers, the age at smoking onset was 15-19 years of age, and the main motivations to start smoking were selfinitiative and influence of friends. The conceptualization of smoking as an illness was significantly higher among the nonsmokers. In 70.6% of the smokers, tobacco intake was 1-10 cigarettes a day. Among the smokers, 92.3% agreed that smoking is harmful to health, 67.3% had tried to quit smoking, 96.0% believed themselves able to do so, and 87.2% intended to quit smoking. CONCLUSIONS: The prevalence of smoking remains significant among medical students. Therefore, it is fundamental that we develop more effective strategies for smoking prevention and cessation in order to reduce the number of smokers among future doctors. PMID- 19547854 TI - Physical therapy in the immediate postoperative period after abdominal surgery. AB - A series of pulmonary complications can occur after abdominal surgery. Therefore, it is necessary to introduce appropriate treatment early in order to minimize postoperative complications. The objective of the present study was to evaluate patients submitted to abdominal surgery in terms of the effect of physical therapy in the immediate postoperative period. This was a randomized clinical trial, in which one group of patients was submitted to physical therapy in the postoperative recovery room and, subsequently, in the infirmary, whereas another group was submitted to physical therapy in the infirmary exclusively. We conclude that physical therapy performed in the immediate postoperative period minimizes losses in lung function and respiratory muscle strength, as well as shortening recovery room stays. PMID- 19547856 TI - Guided sputum sample collection and culture contamination rates in the diagnosis of pulmonary TB. AB - A comparative study to evaluate contamination in cultures of morning sputum samples, comparing those collected at home under currently recommended conditions and those collected under supervision after patient orientation and education. The home and supervised collection groups produced 43 and 76 sputum samples, respectively. The contamination rate was nearly 3-times higher among samples collected at home than among those collected under supervision (37% vs. 13%, p < 0.05; OR = 0.25). The simple educational and hygiene measures described can decrease the contamination rate among sputum samples collected for diagnostic culture. PMID- 19547855 TI - Health team training and active community surveillance: strategies for the detection of TB cases. AB - OBJECTIVE: To evaluate the impact that Family Health Program (FHP) team training and active surveillance have on the detection of TB cases in a low-income community in the city of Fortaleza, Brazil. METHODS: The study was performed in an area with approximately 25,000 inhabitants, served by a health care center with five FHP teams, in the city of Fortaleza, Brazil. Although all of the teams were trained, active surveillance was carried out only in one of the areas (area 5). We compared the number of TB cases detected in each of the five areas prior to and after the intervention. We also compared the number of TB cases detected in area 5 to the number of those detected in the other areas, as well as to the citywide number of reported TB cases in Fortaleza, within the same period. RESULTS: The number of TB cases detected in the area studied increased from 1 in 2002 to 22 in 2004 (p < 0.05). There was no significant difference between the number of TB cases in area 5 and that observed for each of the other four areas (p > 0.05). There was a significant increase in the number of TB cases detected in the area studied when compared to the city as a whole (p < 0.05). CONCLUSIONS: Training and sensitization of FHP professionals were effective in promoting an increase in the number of TB cases detected in a low-income community. PMID- 19547857 TI - How to prevent, recognize and diagnose infection with the swine-origin Influenza A (H1N1) virus in humans. AB - In March of 2009, a flu epidemic began in Mexico. Shortly thereafter, similar cases appeared in other countries, alerting authorities to the risk of a pandemic. This article details the principal signs and symptoms of infection with the swine-origin Influenza A (H1N1) virus. In addition, the measures to be taken in suspected or confirmed cases are addressed, as are the procedures to follow in relation to contacts. Furthermore, the drugs used in the prophylaxis against and the treatment of infection with the H1N1 virus are described. PMID- 19547858 TI - Avian influenza: the threat of the 21st century. AB - This study aimed to review the literature on infection with the H5N1 subtype of avian influenza A virus, taking into consideration the fact that, in the event of a pandemic, children might become a major risk group. Searches were limited to the past ten years and were carried out using the following electronic databases: Medline, MD Consult, HighWire and Medscape. Children and young adults account for a significant proportion of the susceptible population. We found that more than half of the individuals infected were under 20 years of age and that one quarter was under the age of 10. The incubation period ranged from 2 to 5 days. Initial clinical manifestations are nonspecific, which hinders the diagnosis. Most of the infected individuals presented severe pneumonia, which evolved to respiratory insufficiency within an average of 4 days. Chest X-rays can reveal diffuse multifocal/ interstitial infiltrates or segmental/lobar consolidation with air bronchogram. The pathogenic potential is high, with mortality rates up to 63%, indicating that the pandemic virus might present high pathogenicity and high mortality. Knowledge of the risk of a pandemic and of the measures to be taken in suspect cases constitutes an important step toward controlling a potential pandemic. PMID- 19547859 TI - Aspergilloma formation in cavitary sarcoidosis. AB - Pulmonary cavitation is rather uncommon in patients with sarcoidosis, and aspergilloma is even more uncommon in such cases. Here, we present the case of a 63-year-old female patient with cavitary lung disease who had been under treatment for TB for 9 months. A diagnosis of pulmonary sarcoidosis was established based on the fiberoptic bronchoscopy finding of noncaseating granuloma. Treatment with corticosteroids led to a dramatic improvement in symptoms. While under treatment for sarcoidosis, the patient developed an aspergilloma. She presented immediate skin test reactivity to Aspergillus fumigatus, as well as positivity for A. fumigatus serum precipitins. This is the first reported case of aspergilloma formation in a patient with cavitary sarcoidosis in India. PMID- 19547860 TI - Pseudotumoral presentation of chronic pulmonary schistosomiasis without pulmonary hypertension. AB - Schistosomiasis is a disease that, in Brazil, is caused by Schistosoma mansoni and is transmitted by snails of the genus Biomphalaria. This species occurs in Africa, the Antilles and South America. The parasite, developing to the adult stage in the vascular system of the host, migrates to the mesenteric veins. Each female lays approximately 400 eggs per day, and these travel from the submucosa to the intestinal lumen. There have been reported cases in which S. mansoni eggs were identified in the lungs of patients with no evidence of liver fibrosis. We report a case with this atypical presentation of the disease. The patient presented nonpleuritic chest pain, significant weight loss and dry cough. A CT scan revealed an irregular tumor in the left lower lobe. However, a lung biopsy revealed non-viable S. mansoni eggs. The patient presented no clinical signs of pulmonary or portal hypertension; nor was either identified through diagnostic tests. PMID- 19547862 TI - Environmental control and prevention of respiratory allergy: evidence and obstacles. PMID- 19547861 TI - Diffuse idiopathic pulmonary neuroendocrine cell hyperplasia accompanied by airflow obstruction. AB - Diffuse idiopathic pulmonary neuroendocrine cell hyperplasia with airflow obstruction is a rare form of lung injury. All of the reported cases have been diagnosed by surgical lung biopsy. Only three of the reported cases presented with diffuse interstitial lung opacities on HRCT scans. We report three additional cases of this entity. All of the patients were female and presented with mild-to-moderate airflow obstruction. In the first case, transbronchial biopsy and imaging data were sufficient to make the diagnosis. Although the HRCT scans of all three cases revealed a mosaic pattern, that of the third patient also revealed diffuse interstitial infiltrate. In extremely rare cases, HRCT findings can simulate those seen in other interstitial lung diseases. PMID- 19547864 TI - HSV-1-derived amplicon vectors: recent technological improvements and remaining difficulties--a review. AB - Amplicons are defective and non-integrative vectors derived from herpes simplex virus type 1. As the vector genome carries no virus genes, amplicons are both non toxic for the infected cells and non-pathogenic for the inoculated organisms. In addition, the large transgenic capacity of amplicons, which allow delivery of up to 150 Kbp of foreign DNA, makes these vectors one of the most powerful, interesting and versatile gene delivery platforms. We present here recent technological developments that have significantly improved and extended the use of amplicons, both in cultured cells and in living organisms. In addition, this review also discusses the many difficulties still pending to be solved, in order to achieve stable and physiologically regulated transgene expression. PMID- 19547865 TI - Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis. AB - To improve the serodiagnosis of human toxocariasis, a sensitive and specific enzyme-linked immunoelectrotransfer blot (EITB-IgG) test was developed and evaluated using Toxocara canislarvae excretory-secretory antigens for detecting anti-Toxocara IgG antibodies. The EITB-IgG profile of toxocariasis was characterized by comparing 27 sera from patients with toxocariasis, 110 sera from healthy subjects and 186 sera from patients with other helminth diseases (ascariasis, ancylostomiasis, trichuriasis, enterobiasis, strongyloidiasis, hymenolepiasis, diphyllobothriasis, taeniasis, cysticercosis, hydatidosis and fascioliasis). Antigenic bands of 24, 28, 30, 35, 56, 117, 136 and 152 kDa were predominantly recognized in sera from all patients with toxocariasis. However, only bands of 24-35 kDa were highly specific for Toxocara infection (98.3%), whereas other antigenic bands observed displayed cross-reactivity. Additionally, when the results of the EITB-IgG test were compared to those of the ELISA-IgG test, a 100% concordance was observed for positive results in human toxocariasis cases. The concordance for negative results between the two tests for healthy subjects and patients with other helminth diseases were 96.3% and 53.7%, respectively, showing that the EITB-IgG test has a higher specificity than ELISA. In conclusion, the EITB-IgG test is a very useful tool to confirm the serological diagnosis of human toxocariasis. PMID- 19547866 TI - The Pneumocystis life cycle. AB - First recognised as 'schizonts' of Trypanosoma cruzi, Pneumocystis organisms are now considered as part of an early-diverging lineage of Ascomycetes. As no robust long-term culture model is available, most data on the Pneumocystis cell cycle have stemmed from ultrastructural images of infected mammalian lungs. Although most fungi developing in animals do not complete a sexual cycle in vivo, Pneumocystis species constitute one of a few exceptions. Recently, the molecular identification of several key players in the fungal mating pathway has provided further evidence for the existence of conjugation and meiosis in Pneumocystisorganisms. Dynamic follow-up of stage-to-stage transition as well as studies of stage-specific proteins and/or genes would provide a better understanding of the still hypothetical Pneumocystislife cycle. Although difficult to achieve, stage purification seems a reasonable way forward in the absence of efficient culture systems. This mini-review provides a comprehensive overview of the historical milestones leading to the current knowledge available on the Pneumocystis life cycle. PMID- 19547867 TI - Population structure and diversity of the pathogenic fungus Aspergillus fumigatus isolated from different sources and geographic origins. AB - Fifty-five clinical and environmental Aspergillus fumigatus isolates from Mexico, Argentina, France and Peru were analyzed to determine their genetic variability, reproductive system and level of differentiation using amplified fragment length polymorphism markers. The level of genetic variability was assessed by measuring the percentage of polymorphic loci, number of effective alleles, expected heterozygocity and by performing an association index test (I(A)). The degree of genetic differentiation and variation was determined using analysis of molecular variance at three levels. Using the paired genetic distances, a dendrogram was built to detect the genetic relationship among alleles. Finally, a network of haplotypes was constructed to determine the geographic relationship among them. The results indicate that the clinical isolates have greater genetic variability than the environmental isolates. The I(A) of the clinical and environmental isolates suggests a recombining population structure. The genetic differentiation among isolates and the dendrogram suggest that the groups of isolates are different. The network of haplotypes demonstrates that the majority of the isolates are grouped according to geographic origin. PMID- 19547868 TI - Congenital toxoplasmosis: evaluation of serological methods for the detection of anti-Toxoplasma gondii IgM and IgA antibodies. AB - A study was carried out to evaluate the presence of serological markers for the immunodiagnosis of the vertical transmission of toxoplasmosis. We tested the sensitivity, specificity and predictive values (positive and negative) of different serological methods for the early diagnosis of congenital toxoplasmosis. In a prospective longitudinal study, 50 infants with suspected congenital toxoplasmosis were followed up in the ambulatory care centre of Congenital Infections at University Hospital in Goiania, Goias, Brazil, from 1 January 2004-30 September 2005. Microparticle Enzyme Immunoassay (MEIA), Enzyme Linked Fluorescent Assay (ELFA) and Immune-Fluorescent Antibody Technique (IFAT) were used to detect specific IgM anti-Toxoplasma gondii antibodies and a capture ELISA was used to detect specific IgA antibodies. The results showed that 28/50 infants were infected. During the neonatal period, IgM was detected in 39.3% (11/28) of those infected infants and IgA was detected in 21.4% (6/28). The sensitivity, specificity and predictive values (positive and negative) of each assay were, respectively: MEIA and ELFA: 60.9%, 100%, 100%, 55.0%; IFAT: 59.6%, 91.7%, 93.3%, 53.7%; IgA capture ELISA: 57.1%, 100%, 100%, 51.2%. The presence of specific IgM and IgA antibodies during the neonatal period was not frequent, although it was correlated with the most severe cases of congenital transmission. The results indicate that the absence of congenital disease markers (IgM and IgA) in newborns, even after confirming the absence with several techniques, does not constitute an exclusion criterion for toxoplasmosis. PMID- 19547869 TI - RFLP clusters of Mycobacterium tuberculosis strains from the Indian Ocean Region: local and South Asian characteristics. AB - This is the first study describing the genetic polymorphism of Mycobacterium tuberculosis strains in the Indian Ocean Region. Using IS6110 RFLP analysis, 475 M. tuberculosis isolates from Madagascar, Comoros, Mauritius, Mozambique and La Reunion were compared. Of the 332 IS6110 profiles found, 43 were shared by clusters containing 2-65 strains. Six clusters were common to at least two countries. Of 52 families of strains with similar IS6110 profiles, 10 were common to at least two countries. Interestingly, another characteristic was the frequency (16.8%) of IS6110 single-copy strains. These strains could be distinguished using the DR marker. This preliminary evaluation suggests genetic similarity between the strains of the Indian Ocean Region. However, additional markers would be useful for epidemiological studies and to assess the ancient transmission of strains between countries of this region. PMID- 19547870 TI - Echovirus 30 associated with cases of aseptic meningitis in state of Para, Northern Brazil. AB - Investigation of the aetiology of viral meningitis in Brazil is most often restricted to cases that occur in the Southern and Southeastern Regions; therefore, the purpose of this study is to describe the viral meningitis cases that occurred in state of Para, Northern Brazil, from January 2005-December 2006. The detection of enterovirus (EV) in cerebrospinal fluid was performed using cell culture techniques, RT-PCR, nested PCR and nucleotide sequencing. The ages of the 91 patients ranged from < one year old to > 60 years old (median age 15.90 years). Fever (87.1%), headache (77.0%), vomiting (61.5%) and stiffness (61.5%) were the most frequent symptoms. Of 91 samples analyzed, 18 (19.8%) were positive for EV. Twelve were detected only by RT- PCR followed by nested PCR, whereas six were found by both cell culture and RT-PCR. From the last group, five were sequenced and classified as echovirus 30 (Echo 30). Phylogenetic analyses revealed that Echo 30 detected in Northern Brazil clustered within a unique group with a bootstrap value of 100% and could constitute a new subgroup (4c) according to the phylogenetic tree described by Oberste et al. (1999). This study described the first molecular characterization of Echo 30 in Brazil and this will certainly contribute to future molecular analyses involving strains detected in other regions of Brazil. PMID- 19547871 TI - The IFN-gamma +874T/A gene polymorphism is associated with retinochoroiditis toxoplasmosis susceptibility. AB - Toxoplasmosis is a worldwide zoonosis that generally produces an asymptomatic infection. In some cases, however, toxoplasmosis infection can lead to ocular damage. The immune system has a crucial role in both the course of the infection and in the evolution of toxoplasmosis disease. In particular, IFN-gamma plays an important role in resistance to toxoplasmosis. Polymorphisms in genes encoding cytokines have been shown to have an association with susceptibility to parasitic diseases. The aim of this work was to analyse the occurrence of polymorphisms in the gene encoding IFN-gamma (+874T/A) among Toxoplasma gondii seropositive individuals, including those with ocular lesions caused by the parasite, from a rural population of Santa Rita de Cassia, Barra Mansa, state of Rio de Janeiro, Brazil. Further, we verified which of these polymorphisms could be related to susceptibility to the development of ocular toxoplasmosis. This study included 34 individuals with ocular toxoplasmosis (ocular group) and 134 without ocular lesions (control group). The differences between A and T allele distributions were not statistically significant between the two groups. However, we observed that a higher frequency of individuals from the ocular group possessed the A/A genotype, when compared with the control group, suggesting that homozygocity for the A allele could enhance susceptibility to ocular toxoplasmosis in T. gondii infection. PMID- 19547872 TI - A simple approach improving the performance of urine reagent strips for rapid diagnosis of urinary schistosomiasis in Nigerian schoolchildren. AB - In Nigeria, schistosomiasis, caused predominantly by the species Schistosoma haematobium, is highly endemic in resource-poor communities. We performed a school-based survey in two rural communities in Osun State (Southwestern Nigeria) and assessed macrohaematuria, microhaematuria and proteinuria as indirect indicators for the presence of disease. Urine samples were inspected macroscopically for haematuria and screened for microhaematuria and proteinuria using urine reagent strips. The microscopic examination of schistosome eggs was used as the gold standard for diagnosis. In total, 447 schoolchildren were included in this study and had a 51% prevalence of urinary schistosomiasis. The sensitivity of microhaematuria (68%) and proteinuria (53%) for infection with S. haematobium was relatively low. In patients with a heavy infection (>or= 500 eggs/10 mL), the sensitivity of microhaematuria was high (95%). When the presence of macrohaematuria and the concomitant presence of microhaematuria and proteinuria were combined, it revealed a sensitivity of 63%, a specificity of 93% and a positive predictive value of 91%. Macrohaematuria also showed high specificity (96%) and a positive predictive value of 92%, while sensitivity was < 50%. These data show that combining urine reagent strip tests (presence of proteinuria and microhaematuria) and information on macrohaematuria increased the accuracy of the rapid diagnosis of urinary schistosomiasis in an endemic rural West African setting. This simple approach can be used to increase the quality of monitoring of schistosomiasis in schoolchildren. PMID- 19547873 TI - Effectiveness and safety of isoniazid chemoprophylaxis for HIV-1 infected patients from Rio de Janeiro. AB - The clinical and epidemiological characteristics, adverse events, treatment adherence and effectiveness of isoniazid chemoprophylaxis were analyzed in a cohort of 138 tuberculosis/HIV-coinfected patients. An open, non-randomized, pragmatic prophylactic trial was conducted on adult patients with a normal chest X-ray and positive tuberculin skin test (>or= 5 mm) who received isoniazid chemoprophylaxis (300 mg/day) for six months. The mean of follow up was 2.8 years (SD 1.3). Adherence to chemoprophylaxis was 87.7% (121/138). Only one patient presented tuberculosis after the end of chemoprophylaxis, corresponding to 0.3 cases per 100 persons per year. The relative risk of some adverse effects was 4.6 times higher (95% CI: 1.9-11.5) in patients with positive anti-HCV serology (4/9, 44.4%) compared to those with negative serology (12/129, 9.6%) (p = 0.002). This study provides evidence regarding the effectiveness and safety of a short and self-administered isoniazid regimen. We recommend the implementation of this routine by health service practitioners. PMID- 19547874 TI - Mutations in rpoB and katG genes in Mycobacterium isolates from the Southeast of Mexico. AB - The most frequent mutations associated with rifampin and isoniazid resistance in Mycobacterium are the substitutions at codons 531 and 315 in the rpoB and katG genes, respectively. Hence, the aim of this study was to characterize these mutations in Mycobacterium isolates from patients suspected to be infected with drug-resistant (DR) pulmonary tuberculosis (TB) in Veracruz, Mexico. Drug susceptibility testing of 25 clinical isolates revealed that five were susceptible while 20 (80%) were DR (15% of the annual prevalence for Veracruz). Of the DR isolates, 15 (75%) were resistant to rifampin, 17 (85%) to isoniazid and 15 (75%) were resistant to both drugs (MDR). Sequencing analysis performed in the isolates showed that 14 (93%) had mutations in the rpoB gene; seven of these (47%) exhibited a mutation at 531 (S-->L). Ten (58%) of the 20 resistant isolates showed mutations in katG; nine (52%) of these 10 exhibited a mutation at 315 (S- >T). In conclusion, the DR profile of the isolates suggests a significant number of different DR-TB strains with a low frequency of mutation at codons 531 and 315 in rpoB and katG, respectively. This result leads us to consider different regions of the same genes, as well as other genes for further analysis, which is important if a genetic-based diagnosis of DR-TB is to be developed for this region. PMID- 19547875 TI - Cl gene cluster encoding several small nucleolar RNAs: a comparison amongst trypanosomatids. AB - Small nucleolar RNAs (snoRNAs) are small non-coding RNAs that modify RNA molecules such as rRNA and snRNA by guiding 2'-O-ribose methylation (C/D box snoRNA family) and pseudouridylation reactions (H/ACA snoRNA family). H/ACA snoRNAs are also involved in trans-splicing in trypanosomatids. The aims of this work were to characterise the Cl gene cluster that encodes several snoRNAs in Trypanosoma rangeli and compare it with clusters from Trypanosoma cruzi, Trypanosoma brucei, Leishmania major, Leishmania infantum, Leishmania braziliensis and Leptomonas collosoma. The T. rangeli Cl gene cluster is an 801 base pair (bp) repeat sequence that encodes three C/D (Cl1, Cl2 and Cl4) and three H/ACA (Cl3, Cl5 and Cl6) snoRNAs. In contrast to T. brucei, the Cl3 and Cl5 homologues have not been annotated in the Leishmania or T. cruzi genome projects (http//:www.genedb.org). Of note, snoRNA transcribed regions have a high degree of sequence identity among all species and share gene synteny. Collectively, these findings suggest that the Cl cluster could constitute an interesting target for therapeutic (gene silencing) or diagnostic intervention strategies (PCR derived tools). PMID- 19547876 TI - Response of Triatoma infestans to pour-on cypermethrin applied to chickens under laboratory conditions. AB - This article reports the effects of a pour-on formulation of cypermethrin (6% active ingredient) applied to chickens exposed to Triatoma infestans, the main vector of Chagas disease in rural houses of the Gran Chaco Region of South America. This study was designed as a completely random experiment with three experimental groups and five replicates. Third instar nymphs were fed on chickens treated with 0, 1 and 2 cc of the formulation. Nymphs were allowed to feed on the chickens at different time intervals after the insecticide application. Third instar nymphs fed on treated chickens showed a higher mortality, took less blood during feeding and had a lower moulting rate. The mortality rate was highest seven days after the insecticide solution application and blood intake was affected until 30 days after the application of the solution. PMID- 19547877 TI - Preferential transcription of Paracoccidioides brasiliensis genes: host niche and time-dependent expression. AB - Paracoccidioides brasiliensis causes infection through inhalation by the host of airborne propagules from the mycelium phase of the fungus. This fungus reaches the lungs, differentiates into the yeast form and is then disseminated to virtually all parts of the body. Here we review the identification of differentially-expressed genes in host-interaction conditions. These genes were identified by analyzing expressed sequence tags (ESTs) from P. brasiliensis cDNA libraries. The P. brasiliensis was recovered from infected mouse liver as well as from fungal yeast cells incubated in human blood and plasma, mimicking fungal dissemination to organs and tissues and sites of infection with inflammation, respectively. In addition, ESTs from a cDNA library of P. brasiliensis mycelium undergoing the transition to yeast were previously analyzed. Together, these studies reveal significant changes in the expression of a number of genes of potential importance in the host-fungus interaction. In addition, the unique and divergent representation of transcripts when the cDNA libraries are compared suggests differential gene expression in response to specific niches in the host. This analysis of gene expression patterns provides details about host-pathogen interactions and peculiarities of sites within the host. PMID- 19547878 TI - The interaction between Histoplasma capsulatum cell wall carbohydrates and host components: relevance in the immunomodulatory role of histoplasmosis. AB - Histoplasma capsulatum is an intracellular fungal pathogen that causes respiratory and systemic disease by proliferating within phagocytic cells. The binding of H. capsulatum to phagocytes may be mediated by the pathogen's cell wall carbohydrates, glucans, which consist of glucose homo and hetero-polymers and whose glycosydic linkage types differ between the yeast and mycelial phases. The alpha-1,3-glucan is considered relevant for H. capsulatum virulence, whereas the beta-1,3-glucan is antigenic and participates in the modulation of the host immune response. H. capsulatum cell wall components with lectin-like activity seem to interact with the host cell surface, while host membrane lectin-like receptors can recognize a particular fungal carbohydrate ligand. This review emphasizes the relevance of the main H. capsulatum and host carbohydrate-driven interactions that allow for binding and internalization of the fungal cell into phagocytes and its subsequent avoidance of intracellular elimination. PMID- 19547879 TI - Inteins in pathogenic fungi: a phylogenetic tool and perspectives for therapeutic applications. AB - Inteins or 'internal proteins' are coding sequences that are transcribed and translated with flanking sequences (exteins). After translation, the inteins are excised by an autocatalytic process and the host protein assumes its normal conformation and develops its expected function. These parasitic genetic elements have been found in important, conserved proteins in all three domains of life. Most of the eukaryotic inteins are present in the fungi kingdom and the PRP8 intein is one of the most widespread inteins, occurring in important pathogens such as Cryptococcus neoformans (varieties grubii and neoformans), Cryptococcus gattii, Histoplasma capsulatum and Paracoccidioides brasiliensis. The knowledge of conserved and non-conserved domains in inteins have opened up new opportunities for the study of population variability in pathogenic fungi, including their phylogenetic relationships and recognition or diagnoses of species. Furthermore, inteins in pathogenic fungi should also be considered a promising therapeutic drug target, since once the autocatalytic splicing is inhibited, the host protein, which is typically vital, will not be able to perform its normal function and the fungal cell will not survive or reproduce. PMID- 19547880 TI - Phylogeny and evolution of the aspartyl protease family from clinically relevant Candida species. AB - Aspartyl proteases are a class of enzymes that include the yeast aspartyl proteases and secreted aspartyl protease (Sap) superfamilies. Several Sap superfamily members have been demonstrated or suggested as virulence factors in opportunistic pathogens of the genus Candida. Candida albicans, Candida tropicalis, Candida dubliniensis and Candida parapsilosis harbour 10, four, eight and three SAP genes, respectively. In this work, genome mining and phylogenetic analyses revealed the presence of new members of the Sap superfamily in C. tropicalis (8), Candida guilliermondii (8), C. parapsilosis(11) and Candida lusitaniae (3). A total of 12 Sap families, containing proteins with at least 50% similarity, were discovered in opportunistic, pathogenic Candida spp. In several Sap families, at least two subfamilies or orthologous groups were identified, each defined by > 90% sequence similitude, functional similarity and synteny among its members. No new members of previously described Sap families were found in a Candida spp. clinical strain collection; however, the universality of SAPT gene distribution among C. tropicalis strains was demonstrated. In addition, several features of opportunistic pathogenic Candida species, such as gene duplications and inversions, similitude, synteny, putative transcription factor binding sites and genome traits of SAP gene superfamily were described in a molecular evolutionary context. PMID- 19547881 TI - Mortality due to systemic mycoses as a primary cause of death or in association with AIDS in Brazil: a review from 1996 to 2006. AB - Deaths caused by systemic mycoses such as paracoccidioidomycosis, cryptococcosis, histoplasmosis, candidiasis, aspergillosis, coccidioidomycosis and zygomycosis amounted to 3,583 between 1996-2006 in Brazil. When analysed as the underlying cause of death, paracoccidioidomycosis represented the most important cause of deaths among systemic mycoses (approximately 51.2%). When considering AIDS as the underlying cause of death and the systemic mycoses as associated conditions, cryptococcosis (50.9%) appeared at the top of the list, followed by candidiasis (30.2%), histoplasmosis (10.1%) and others. This mortality analysis is useful in understanding the real situation of systemic mycoses in Brazil, since there is no mandatory notification of patients diagnosed with systemic mycoses in the official health system. PMID- 19547882 TI - Prevalence of antiretroviral drug resistance among treatment-naive and treated HIV-infected patients in Venezuela. AB - An in-house, low-cost method was developed to determine the genotypic resistance of immunodeficiency virus type 1 (HIV-1) isolates. All 179 Venezuelan isolates analysed belonged to subtype B. Primary drug resistance mutations were found in 11% of 63 treatment-naive patients. The prevalence of resistance in isolates from 116 HIV-positive patients under antiretroviral treatment was 47% to protease inhibitors, 65% to nucleoside inhibitors and 38% to non-nucleoside inhibitors, respectively. Around 50% of patients in the study harboured viruses with highly reduced susceptibility to the three classical types of drugs after only five years from their initial diagnoses. PMID- 19547884 TI - [Obesity and pregnancy]. PMID- 19547883 TI - Dengue virus type 3 in Brazil: a phylogenetic perspective. AB - Circulation of a new dengue virus (DENV)-3 genotype was recently described in Brazil and Colombia, but the precise classification of this genotype has been controversial. Here we perform phylogenetic and nucleotide-distance analyses of the envelope gene, which support the subdivision of DENV-3 strains into five distinct genotypes (GI to GV) and confirm the classification of the new South American genotype as GV. The extremely low genetic distances between Brazilian GV strains and the prototype Philippines/L11423 GV strain isolated in 1956 raise important questions regarding the origin of GV in South America. PMID- 19547886 TI - [Depression and anxiety in menopausal women: associated factors]. AB - PURPOSE: to determine the prevalence of depression and anxiety in climacteric women and the probable factors responsible for its occurrence. METHODS: a transversal study that has selected 93 women attended at a climacteric outpatient clinic, from May 2006 to August 2007. Inclusion criteria were: women from 40 to 65 years old who agreed with participating in the project. EXCLUSION CRITERIA: patients in hormonal therapy, hormone-therapy by implant, DIUs and depo injections in the preceding six months, endocrinopathies leading to menstrual irregularities, hepatopathies, thrombopathies, use of drugs which interfere in the menstrual cycle, anxiolytics and antidepressants (as their use indicates previous diagnosis of mood disorders), hysterectomy, oophorectomy, cancer or psychiatric disease, and patients who had been submitted to radio or chemotherapy. During the interview, four questionnaires were applied: Anamnesis, containing socio-demographic, clinical and living habits data; Blatt-Kupperman's Menopausal Index for climacteric syndrome diagnosis; Anxiety sub-scale of the Hospital Anxiety and Depression scale (HADS-A) for anxiety diagnosis; and Beck's Depression Inventory for the diagnosis of depression. Descriptive and correlation analysis among the variables, chi2 and Hosmer-Lemeshow tests were performed using the Statistica Software program, version 6. RESULTS: the average depression prevalence among the patients was 36.8%, while that of anxiety was 53.7%. There was no significant difference between the prevalence of depression and anxiety in the three phases of climacterium. There was a significant relationship between the presence of moderate climacteric symptoms and the presence of mood alterations (p<0.001). Depression was more frequent in women with anxiety (OR=4.2) and insomnia (OR=4.9), having a job being a protection factor (OR=0.2). Risk factors related to anxiety were the presence of depression (OR=6.1) and antecedents of pre-menstrual tension (OR=7.0). CONCLUSIONS: the prevalence of depression and anxiety is high in climacterium, being possible to detect risk factors related to their occurrence. PMID- 19547885 TI - [Cardiovascular risk markers in polycystic ovary syndrome in women with and without insulin resistance]. AB - PURPOSE: to evaluate whether the presence of insulin resistance (IR) alters cardiovascular risk factors in women with polycystic ovary syndrome (POS). METHODS: transversal study where 60 POS women with ages from 18 to 35 years old, with no hormone intake, were evaluated. IR was assessed through the quantitative insulin sensitivity check index (QUICKI) and defined as QUICKI <0.33. The following variables have been compared between the groups with or without IR: anthropometric (weight, height, waist circumference, arterial blood pressure, cardiac frequency), laboratorial (homocysteine, interleucines-6, factor of tumoral-alpha necrosis, testosterone, fraction of free androgen, total cholesterol and fractions, triglycerides, C reactive protein, insulin, glucose), and ultrasonographical (distensibility and carotid intima-media thickness, dilation mediated by the brachial artery flux). RESULTS: Eighteen women (30%) presented IR and showed significant differences in the following anthropometric markers, as compared to the women without IR (POS with and without IR respectively): body mass index (35.56+/-5.69 kg/m(2) versus 23.90+/-4.88 kg/m(2), p<0.01), waist (108.17+/-11.53 versus 79.54+/-11.12 cm, p<0.01), systolic blood pressure (128.00+/-10.80 mmHg versus 114.07+/-8.97 mmHg, p<0.01), diastolic blood pressure (83.67+/-9.63 mmHg versus 77.07+/-7.59 mmHg, p=0.01). It has also been observed significant differences in the following laboratorial markers: triglycerides (120.00+/-56.53 mg/dL versus 77.79+/-53.46 mg/dL, p=0.01), HDL (43.06+/-6.30 mg/dL versus 40.45+/-10.82 mg/dL, p=0.01), reactive C protein (7.98+/-10.54 mg/L versus 2.61+/-3.21 mg/L, p<0.01), insulin (28.01+/-18.18 microU/mL versus 5.38+/-2.48 microU/mL, p<0.01), glucose (93.56+/-10.00 mg/dL versus 87.52+/-8.75 mg/dL, p=0.02). Additionally, two out of the three ultrasonographical markers of cardiovascular risk were also different between the groups: carotid distensibility (0.24+/-0.05 mmHg-1 versus 0.30+/-0.08 mmHg-1, p<0.01) and carotid intima-media thickness (0.52+/-0.08 mm versus 0.43+/-0.09, p<0.01). Besides, the metabolic syndrome ratio was higher in women with IR (nine cases=50% versus three cases=7.1%, p<0.01). CONCLUSIONS: POS and IR women present significant differences in several ultrasonographical, seric and anthropometric markers, which point out to higher cardiovascular risk, as compared to women without POS and IR. In face of that, the systematic IR evaluation in POS women may help to identify patients with cardiovascular risk. PMID- 19547887 TI - [Effect of tibolone on endometrium of castrated rats]. AB - PURPOSE: to evaluate the effect of long-term use of a high dose of tibolone on the morphology of the endometrium of castrated female rats. METHODS: fifteen female Wistar rats, aged eight weeks and weighting about 250 g were used. All the female rats were submitted to bilateral oophorectomy and 30 days afterwards, vaginal cytology was collected, to verify the menopause status. The female rats were randomly divided in two groups. The Treatment Group (n=9) received 1 mg of tibolone/day orally; the Control Group (n=6) received a solution of carboxymethylcellulose vehicle. After 20 weeks of treatment, all the animals were sedated and sacrificed by cervical dislocation. The uterus was removed and fixated in 10% buffer formaldehyde. Both uterine horns were divided in three regions (proximal, medial and distal) and processed to be included in paraffin. Histological sections, stained with hematoxylin-eosin were submitted to morphological and morphometrical analysis. The following parameters have been analyzed: thickness of the endometrial superficial epithelium, thickness of the endometrium stroma, endometrial area, absolute number of endometrial glands and number of glands/endometrial area. The data obtained were compared by the t Student test. RESULTS: in the Tibolone Group, the uteri were well developed and there was a significant increase (p<0.01) of all the histomorphometric parameters. In some cases, the cylindrical epithelium became stratified, pavimentous and covered the internal portions of the glands, as well as of the endometrium cavity. Rats from the Control Group presented uterine atrophy. There were few tubular-like glands and scarce intercellular substance. Glands were covered by cubic epithelium which extended itself to the endometrial cavity. CONCLUSIONS: high doses of tibolone, given for long periods of time to castrated female rats, have an estrogenic effect which can be dose-dependent, causing proliferation in the endometrium and causing changes in the cell differentiation (squamous metaplasia), but do not lead to hyperplasia. PMID- 19547888 TI - [Effects of tamoxifen on the expression of TGF-beta and p27 proteins in polyps and adjacent endometrium in postmenopausal women]. AB - PURPOSE: to evaluate the effects of tamoxifen on the expression of TGF-beta and p27 proteins in polyps and adjacent endometrium of women after menopause. METHODS: prospective study with 30 post-menopausal women with diagnosis of breast cancer, taking tamoxifen (20 mg/day), presenting diagnosis of suspect endometrial polyps through transvaginal ultrasonography, and submitted to diagnostic and surgical hysterectomy to withdraw the polyps and adjacent endometrium. A immunohistochemical study has been done to verify the expression of the TGF-beta and p27 proteins in the polyps and adjacent endometrium. These proteins' quantification has been done by morphometry. RESULTS: the patients' average age was 61.7 years old; their average age at the menopause onset was 49.5; and the average of using tamoxifen was 25.3 months. The average concentration of positive cells for TGF-beta protein in the glandular and stroma polyp epithelium was 62.6+/-4.5 cells/mm(2). For the p27, in the glandular polyp epithelium, it was 24.2+/-18.6 cells/mm(2) and for the stroma, 19.2+/-15.2 cells/mm(2). There was no significant difference between the expression of TGF-beta and p27 in the glandular epithelial form the polyps and the adjacent endometrium. The expression of proteins in the polyp and adjacent endometrium with its respective glandular and stroma epithelium showed a significant difference for the p27 protein (r=0.9, p<0.05). CONCLUSIONS: we have concluded that the TGF-beta expression is not related to the effect of tamoxifen on the growing of endometrial polyps, but the absence of polyps' malignization by tamoxifen may be explained by the high expression of p27 protein in its glandular epithelium. PMID- 19547889 TI - [Spontaneous pregnancies after ovarian puncture for in vitro maturation in women with the polycystic ovary syndrome]. AB - PURPOSE: to report three cases of spontaneous gestation in women with polycystic ovarian syndrome (PCOS), that occurred in the months subsequent to transvaginal oocyte retrieval for in vitro maturation (IVM). METHODS: three infertile patients with PCOS, submitted to oocytes' IVM without previous ovarian stimulation, were included in the study. During the procedure of oocytes' collection, each ovary was drilled from four to eight times. RESULTS: none of the patients got pregnant with the IVM technique. Evaluating the cases' follow-up, in seven months after the procedure, the three patients got pregnant without the help of techniques of assisted reproduction, which resulted in three births. CONCLUSIONS: the multiple drillings in the ovary of these patients with PCOS, during the process to collect oocytes, may have contributed to their pregnancy in the months following the procedure. PMID- 19547890 TI - [Evidence of follicle responsiveness to FSH by antimullerian hormone in ovulating women]. AB - PURPOSE: to test the hypothesis that the anti-mullerian hormone (AMH) serum level reflects the antral follicles' response to the administration of FSH. METHODS: prospective study, including 116 normo-ovulatory infertile patients submitted to controlled ovarian hyperstimulation with GnRH and FSH agonists. The AMH serum level was measured after reaching the pituitary suppression and before the FSH administration (basal day). The number of antral follicles was determined by ultrasonography at the basal day (precocious antral follicles; 2 to 8 mm) and at the day of hCG administration (dhCG; pre-ovulatory follicles; >16 mm). The follicle response to FSH was determined by the percentage of precocious antral follicles which reached pre-ovulatory stage in response to FSH (maturation rate). The correlation of AMH with the patients' age, the total number of precocious antral and pre-ovulatory follicles, collected oocytes, total dose of FHS in the controlled ovarian stimulation and the rate of follicular maturation was studied. For the statistical analysis, it simple regression analysis and the Spearman's test were used, at a 5% significance level. RESULTS: The serum level of AMH was positively correlated with the number of precocious antral follicles at the basal day (r=0.64; p<0.0001) and pre-ovulatory follicles in dhCG (r=0.23; p=0.01). Exceptionally, the serum level of AMH was negatively correlated with the maturation ratio (r=-0.24; p<0.008). CONCLUSIONS: AMH attenuates the follicular development caused by FSH administration. PMID- 19547891 TI - [Evaluation of the main prenatal routine exams (part 1)]. AB - Good prenatal care is essential to guarantee maternal and perinatal health. Nowadays, with the constant progress on the diagnostic methods and changes in diseases panorama, like increased frequency of diabetes and sexually transmitted diseases, several diagnostic procedures have been studied. These challenges difficult the selection of prenatal exams by the clinicians, in order to improve maternal and perinatal outcome. This review aimed to evaluate the main prenatal routine exams, according to the best current scientific evidences. PMID- 19547894 TI - Open lung biopsy in patients on mechanical ventilation with suspected diffuse lung disease. AB - BACKGROUND: While open lung biopsy (OLB) performed in patients on mechanical ventilation (MV) with diffuse lung diseases (DLD) can be extremely important in establishing the diagnosis, the associated risk of this procedure should be taken into account. AIM: To determine the diagnostic yield, therapeutic changes, complications and mortality in patients with DLD on MV submitted to OLB. METHODS: Retrospective study of 19 patients admitted to S. Joao Hospital between January 1999 and July 2007 (8.5 years). Data analysed included demographic data, ventilation variables before and after biopsy, diagnostic yield, effect on subsequent treatment changes and complications of OLB. Statistical analysis was performed using SPSS 14.0. RESULTS: The mean age of patients was 58+/-16.3 years old and 53% were male. The mean duration of hospital stay in Intensive Care Unit before performing OLB was 13+/-7 days. All biopsies were performed by thoracotomy. The diagnostic yield was 95%. There were no significant differences in partial pressure of arterial oxygen/fraction of inspired oxygen (PaO(2)/ FiO(2)) ratio and the positive end expiratory pressure (PEEP) before and after OLB. Postoperative complications occurred in 4 patients (21%; persistent air leak). Alteration in the diagnosis occurred in 14 patients (74%) and in 8 patients (42%) there was a modification in the treatment regimen. Global mortality was 47% (9 patients) but there were no biopsy-related deaths. CONCLUSION: The high diagnostic yield and the low incidence of complications make OLB a useful procedure in patients on MV with DLD of unknown aetiology. However, early OLB may lead to even better results in some patients. PMID- 19547893 TI - Transthoracic biopsy with core cutting needle (Trucut) for the diagnosis of mediastinal tumors. AB - AIM: To determine the contribution of percutaneous biopsy with core cutting needle (Trucut) in the diagnosis of mediastinal tumours. METHOD: Retrospective review of 56 patients with mediastinal lesions who underwent percutaneous core cutting needle biopsy, oriented but not guided by computer assisted tomography of the thorax, 1999 - 2008. RESULTS: Percutaneous biopsy with core cutting needle provided adequate material in 49/56, with a total positive sample rate of 88%. In 7/56 (12%) cases the material was insufficient to define the diagnosis. Percutaneous core cutting needle biopsy established a specific histological diagnosis in 88% of the patients: 23/56 (41%) lymphomas; 12/56 (21%) thymomas; 5/56 (3%) thymic carcinomas; 3/56 (2%) small cell carcinoma and 1/56 (0.6%) metastatic adenocarcinoma, metastatic squamous cell carcinoma, neuroendocrine primitive carcinoma, plasmocytoma, teratoma and goiter. All patients underwent thoracic X-ray after the procedure. No complications were found in these patients. CONCLUSION: Percutaneous core cutting needle biopsy (Trucut) oriented but not guided by computer assisted tomography of the thorax is an easy and safe procedure which can provide a precise diagnosis in the majority of mediastinal tumours and can prevent the need for exploratory thoracic surgery in cases which are medically treatable or non-resectable. PMID- 19547892 TI - Trends in lung cancer mortality in Portugal (1955-2005). AB - INTRODUCTION: While the rate of smoking and lung cancer mortality has been decreasing in western Europe, there was no decline in lung cancer mortality in Portugal until 1998. AIM: To describe lung cancer mortality trends in Portugal. METHODS: Lung cancer mortality rates (International Disease Classification 10: C33-34) in Portugal 1955- 2005 by gender and 5-year age groupings were provided by the World Health Organization and the National Institute of Statistics. Standard mortality rates (direct method, world population) were calculated for the 35-74, 35-44, 45-54, 55-64 and 65-74 year- old age groups. Joinpoint regression was used to calculate the annual percent change (APC) in mortality and to identify any inflection points. RESULTS: Between 1955 and 2005 we observed a stabilisation in lung cancer mortality in men aged 35-74 years old, varying 3.77%/year (95% confidence interval [95%CI]: 3.53-4.01) in 1955-1986 and 0.15%/year (95%CI: -0.99-0.69) in 1996-2005. We observed negative APC point estimates (with none significantly below zero) in the most recent trends except for the 45-54 age group, where we only noted an APC deceleration since 1981. The mortality increased 1.60%/year (95%CI: 1.40-1.77) in women aged 35- 74 years old 1955-2005. CONCLUSION: In the last two decades we observed a lung cancer mortality stabilisation in males, whereas mortality in females increased continuously. These results place Portugal at the end of the third stage of the smoking epidemic. PMID- 19547895 TI - The association of rhinoconjunctivitis and asthma symptoms in adolescents. AB - AIM: Our study aimed to determine the rate of association of rhinoconjunctivitis and asthma symptoms in adolescents to analyse whether asthma symptoms are more severe and frequent in asthmatics with concomitant allergic rhinitis and assess if adolescents are aware of having rhinoconjunctivitis. METHODS: A cross sectional study, with two components: a study in prevalence and an inter-case study (rhinitis symptoms) with a comparison group (no rhinitis symptoms), based on information from questionnaires applied in phase 3 of ISAAC in Recife in 2002. RESULTS: Associated rhinoconjunctivitis and probable asthma symptoms were observed in 5.1% of adolescents (48/940; CI 95%: 3.8 %-6.6%), probable asthma alone in 10.9% (103/940; CI 95%: 9.1%- 13.1%) and rhinoconjunctivitis alone in 9.7% (91/940; CI 95%: 7.9%-13%). Among the rhinitis-bearing adolescents, almost 81.3% (39/48) had persistent probable asthma and 31.8% (48/151) of asthmatic patients rhinoconjunctivitis. 65.1% (86/132) of adolescents with diagnosed rhinitis were unaware of rhinitis symptoms. CONCLUSIONS: The association of rhinoconjunctivitis and asthma symptoms is frequent and associated to more severe asthma symptoms. Adolescents' unawareness of rhinitis symptoms reflects the underdiagnosis that can result in down-playing the symptoms, and the consequent undertreatment. PMID- 19547896 TI - Effects of aging on lung function. A comparison of lung function in healthy adults and the elderly. AB - INTRODUCTION: The respiratory system changes with age and understanding these changes helps detect and prevent respiratory dysfunctions in the elderly. AIMS: This study compares the respiratory pattern in healthy non-smoker adults and the elderly, using lung function testing and expansion of the chest to confirm the effects of aging on lung function. METHODOLOGY: We used lung function testing to measure forced vital capacity, forced expiratory volume in one second, peak expiratory flow rate and maximum voluntary ventilation. We also measured expansion of the chest. Measurements were taken with subjects resting in the dorsal recumbent position with upper body elevated to 45 masculine. Statistical analysis consisted of the Student T test for independent samples, the non parametric Mann-Whitney test with a p <0.05 level, and linear regression analysis, also with a p <0.05 level, to assess correlation between variables studied and age. Our population consisted of 35 elderly subjects and 35 adults. 15 of the elderly subjects were male (43%) and 20 female (57%). 16 of the adult group were male (46%) and 19 female (54%). The sample was mapped in terms of age, weight, height, abdominal girth and clinical data, to exclude factors which could distort the results. RESULTS: The difference between the two study groups attained statistical significance for all parameters measured. The linear relationship was also significant between age and all parameters and a negative and significant correlation was seen. Expansion of the chest in females was the parameter most inversely correlated with age (60.37%). The lung function testing variable with the greatest difference between the groups was peak expiratory flow rate (35.77% in females and 36.17% in males). CONCLUSIONS: Our results show differences in the respiratory patterns of healthy adults and the elderly, suggesting that age impacts on lung function. Both male and female elderly subjects had lower lung function testing scores than the adult subjects, with this difference more marked in females. PMID- 19547898 TI - [Experimental models in oncology: contribution of cell culture on understanding the biology of cancer]. AB - In the beginning of the 20th century, tissue culture was started with the aim of studying the behaviour of animal cells in normal and stress conditions. The cell study at molecular level depends on their capacity of growing and how they can be manipulated in laboratory. In vitro cell culture allows us the possibility of studying biological key processes, such as growth, differentiation and cell death, and also to do genetic manipulations essential to the knowledge of structure and genes function. Human stem cells culture provides strategies to circumvent other models' deficiencies. It seems that cancer stem cells remain quiescent until activation by appropriated micro-environmental stimulation. Several studies reveal that different cancer types could be due to stem cell malignant transformations. Removal of these cells is essential to the development of more effective cancer therapies for advanced disease. On the other hand, dendritic cells modified in culture may be used as a therapeutic vaccine in order to induce tumour withdraw. PMID- 19547897 TI - [Non-invasive ventilation]. AB - Non-invasive ventilation (NIV) is a technique that delivers mechanical ventilation avoiding side effects and complications associated with endotracheal intubation and invasive mechanical ventilation. This technique has proved to be effective in different types of respiratory failure. In this article, the authors revise the advantages and limitations of NIV, interfaces used and indications in acute and acute-on-chronic respiratory failure. PMID- 19547899 TI - [Isolated mediastinal cystic lymphangioma]. AB - A 59 years old female patient, asymptomatic, with the incidental finding of an ovarian tumor in her routine gynecological evaluation, and during the preoperative examinations it was incidentally found an isolated mediastinal tumor, and then routed to diagnostic evaluation of the lesion, which later proved to be a cystic lymphangioma. The cystic hygroma of the mediastinum is a benign tumor and very infrequent, representing only 0.7 to 4.5% of all mediastinal tumors, and of these, only 1% is exclusively mediastinal in location. The definitive diagnosis is only possible by pathological examination, and the recommended treatment consists of complete surgical resection. Cases are described in isolated reports or series with few patients, and their readiness or synchronicity with other tumors, unknown, and to the best of out knowledge, not reported yet. PMID- 19547900 TI - [Secondary hypertrophic osteoarthropathy to lung cancer: a case report]. AB - The secondary hypertrophic osteoarthropathy is a systemic change that affects the bones, joints and soft tissues and is secondary to any intrathoracic pathology. It is a syndrome of chronic proliferative periostitis of the long bones, clubbing of the fingers of the hands, feet or both, and olyarthritis or polyarthritis. We report one case of hypertrophic osteoarthropathy in a patient with lung mass with bulky diagnostic anatomopathological, adenocarcinoma. PMID- 19547901 TI - [Relapsing pleural effusion and gastric polyposis: a case report]. AB - The authors present the case of a lymphoproliferative disorder (LPD) with pleuro pulmonary involvement. It was a very aggressive primary gastric lymphoma of the MALT subtype, diagnosed in an 86-year-old woman admitted in Pulmonology ward at our Hospital with pleural effusion. Helicobacter pylori search was negative, what is infrequent. The disease was already disseminated at the time of the diagnosis and did not respond to various treatment modalities, what is also an uncommon finding. Clinical presentation, diagnosis and management of this disease are reviewed, pointing out its unique features and comparing the particular aspects of this case with the published literature. The authors stress that LPD is an important diagnosis to be held in mind in the case of a pleural effusion of unknown aetiology. PMID- 19547902 TI - [Tuberculous serositis in patient with systemic lupus erythematosus: case report and literature review]. AB - It is a case report of young female that had diagnostic criteria of Systemic Lupus Erithematosus in activity, according American Rheumatology Association. The patient had fever, anemia, arthritis, cellular casts, positive LE cells, positive antinuclear antibody. She has evolved to bilateral pleural effusion and pericardic effusion that both have been initially attributed to lupus. Due to she has also maintained low fever, sudoresis, loss of weight and a persistent serositis, a thoracocenthesis with pleural biopsy has been done and the result of it has revealed granulomatous chronic pleuritis, diagnosticing pleural tuberculosis. A pericardiocenthesis has also been done. After six months of anti tuberculosis therapy, there was a regression of radiologic imaging. Some concepts referring to tuberculosis and systemic lupus erythematosus are discussed, including symptoms, diagnosis and specific situations. It is emphasized the necessity of early diagnosis and appropriate management of tuberculosis disease in lupic patients, in areas where tuberculosis is endemic. PMID- 19547904 TI - Dissipation of fipronil in water under laboratory conditions. AB - Fipronil belongs to phenyl-pyrazole class of chemical compounds. Its dissipation in water has been studied for 90 days under laboratory conditions at two dose 0.028 (T1) and 0.056 (T2) MUg mL-1. Residues were extracted with 15% dichloromethane in hexane by liquid-liquid partitioning and were estimated by GC ECD system equipped with capillary column. Fipronil was found to persist up to 30 days after application and 100% dissipation was observed after 60 days of application. Dissipation followed a biphasic first order kinetics with half-life values of 19.13 and 20.63 days in water at (T1) and (T2) treatments, respectively. PMID- 19547903 TI - Occupational constrictive bronchiolitis with normal physical, functional and image findings. AB - Constrictive bronchiolitis is characterized by alterations in the walls of membranous and respiratory bronchioles. These changes lead to concentric narrowing or complete obliteration of the airway lumen. Suspicion of possible bronchiolar disorders may arise from clinical, functional, and radiologic findings. However, constrictive bronchiolitis may be present even with normal physical, functional and image findings, which turns the diagnosis difficult. A high index of suspicion is necessary to justify invasive tests that lead to pulmonary biopsy. In this report, we describe a patient with cough and dyspnoea, with normal physical, functional and image findings, whose work-up leaded to the diagnosis of constrictive bronchiolitis. PMID- 19547905 TI - Determination of pesticide residues in honey samples. AB - Protocol for the determination of pesticides residues in honey samples have been standardized using a simple technique of liquid-liquid extraction. The method is sensitive to detect low levels of pesticides in honey. Honey sample was fortified with pesticides, namely, cypermethrin, fenvalerate, alphamethrin, lamba cyhalothrin, endosulfan (alpha, beta and sulfate) and chlorpyrifos. The method of extraction and clean up was optimized and validated in the laboratory. The method was applied to screen six samples of honey locally available for pesticides residues. Recoveries ranged from 60% to 90.6% with RSDs from 2% to 10%. Low recoveries were recorded for alpha and beta -endosulfan in the range of 60%-71%. The LOQs, varied from 0.05 to 1.0 mg kg-1. PMID- 19547906 TI - The epidemiology of osteonecrosis: findings from the GPRD and THIN databases in the UK. AB - We conducted a case-control study to examine osteonecrosis (ON) incidence, patient characteristics, and selected potential risk factors using two health record databases in the UK. Statistically significant risk factors for ON included systemic corticosteroid use, hospitalization, referral or specialist visit, bone fracture, any cancer, osteoporosis, connective tissue disease, and osteoarthritis. INTRODUCTION: The purpose of this case-control study was to examine the incidence of osteonecrosis (ON), patient characteristics, and selected potential risk factors for ON using two health record databases in the UK: the General Practice Research Database and The Health Improvement Network. METHODS: ON cases (n = 792) were identified from 1989 to 2003 and individually matched (age, sex, and medical practice) up to six controls (n = 4,660) with no record of ON. Possible risk factors were considered for inclusion based on a review of published literature. Annual incidence rates were computed, and a multivariable logistic regression model was derived to evaluate selected risk factors. RESULTS: ON of the hip represented the majority of cases (75.9%). Statistically significant risk factors for ON were systemic corticosteroid use in the previous 2 years, hospitalization, referral or specialist visit, bone fracture, any cancer, osteoporosis, connective tissue disease, and osteoarthritis within the past 5 years. Only 4.4% of ON cases were exposed to bisphosphonates within the previous 2 years. CONCLUSIONS: This study provides further perspective on the descriptive epidemiology of ON. Studies utilizing more recent data may further elucidate the understanding of ON key predictors. PMID- 19547907 TI - Effect of ivabradine in dobutamine induced sinus tachycardia in a case of acute heart failure. PMID- 19547908 TI - Interventional EUS for the treatment of pancreatic cancer. AB - Since the curved linear array echoendoscope (linear EUS) was developed in the 1990s, EUS has evolved from EUS imaging, to EUS-guided FNA, and now to EUS-guided fine needle injection (FNI), giving EUS even wider application. This advancement has brought "interventional EUS" into the pancreato-biliary field. Interventional EUS for pancreatic cancer includes delivery of contrast agents, drainage/anastomosis, celiac neurolysis (including ganglion neorolysis), radiofrequency ablation, photodynamic therapy, brachytherapy, and delivery of a growing number of anti-tumor agents. This review will focus on interventional EUS in the treatment of pancreatic cancer. PMID- 19547910 TI - Animal insulin therapy induces a biased insulin antibody response that persists for years after introduction of human insulin. AB - Administration of exogenous insulin for the treatment of diabetes is often accompanied by the development of insulin antibodies (IA). These antibodies may affect the patient's requirement for insulin by acting as an insulin binding reservoir. The improvement of insulin purification in the 1970s and the development of human recombinant insulin both reduced the incidence of IA and their binding levels. This study investigates the parameters affecting IA frequency and binding levels in a cohort of type 1 diabetes (T1D) patients. All patients were treated with human recombinant insulin. About half of the patients had received animal insulin prior to the introduction of human recombinant insulin. We tested the IA frequency and binding level for all serum samples. IA were further analyzed for their epitope specificity comparing human and porcine insulin binding. We found that T1D patients who received animal insulin in the past show significantly higher IA binding levels as compared to patients treated exclusively with human recombinant insulin (IA binding level of 0.9 and 0.25 index, respectively, P = 0.005). T1D patients who received animal insulin in the past showed a relative bias towards porcine insulin, as compared to T1D patients who were treated with human recombinant insulin exclusively (P < 0.0001). We conclude that IA binding level and epitope specificity are biased by treatment with animal insulin. This bias remains for over 20 years after animal insulin treatment is terminated. PMID- 19547909 TI - Immune responses of TLR5(+) lamina propria dendritic cells in enterobacterial infection. AB - Toll-like receptors (TLRs) recognize distinct microbial components and induce innate immune responses. TLR5 has been shown to recognize bacterial flagellin. Unlike other TLRs, TLR5 is not expressed on conventional dendritic cells or macrophages. By contrast, TLR5 is mainly expressed on intestinal CD11c(+) lamina propria cells (LPCs), which do not express TLR4. These cells detect pathogenic bacteria and secreted proinflammatory cytokines, mainly in a TLR5-dependent manner. Notably, transport of pathogenic Salmonella typhimurium from the intestinal tract to mesenteric lymph nodes was impaired in Tlr5 (-/-) mice, suggesting that CD11c(+) LPCs expressing TLR5 are used by S. typhimurium for systemic infection. CD11c(+) LPCs consist of four subsets distinguished by differential expression patterns of CD11c and CD11b. CD11c(hi)CD11b(hi) LPDCs have been identified as TLR5-expressing cells. In response to flagellin, these LPDCs induce the differentiation of naive B cells into IgA(+) plasma cells via a mechanism independent of gut-associated lymphoid tissue (GALT), and trigger the differentiation of antigen-specific Th17 and Th1 cells. These LPDCs have unique properties in that they can induce acquired immunity as well as innate immunity via TLR5 in the intestine. PMID- 19547911 TI - Bovine immunodeficiency virus: identification of a long terminal repeat sequence with enhanced promoter activity. AB - We previously identified a new bovine immunodeficiency virus (BIV) trans activator factor of transcription (Tat236) that was derived from a variant of BIV. Here, we report a new BIV long terminal repeat (LTR) sequence (LTRn) that was obtained by PCR from the DNA of cells infected with the BIV variant mentioned above. Sequence analysis indicated that the LTRn U3 region harbors three nucleic acid mutations at residue positions -194, -135 and -114 when compared to the original (wild-type) LTR sequence. Reporter gene assays indicated that LTRn promotes basal and Tat-mediated transactivation activity to levels significantly higher than those obtained with the wild-type LTR. Restoration experiments to the wild-type genotype indicated that both the -135 and -114 nucleic acid substitutions were responsible for the enhanced promoter activity of BIV LTRn. PMID- 19547914 TI - The current structural and functional understanding of APOBEC deaminases. AB - The apolipoprotein B mRNA-editing enzyme catalytic polypeptide (APOBEC) family of cytidine deaminases has emerged as an intensively studied field as a result of their important biological functions. These enzymes are involved in lipid metabolism, antibody diversification, and the inhibition of retrotransposons, retroviruses, and some DNA viruses. The APOBEC proteins function in these roles by deaminating single-stranded (ss) DNA or RNA. There are two high-resolution crystal structures available for the APOBEC family, Apo2 and the C-terminal catalytic domain (CD2) of Apo3G or Apo3G-CD2 [Holden et al. (Nature 456:121-124, 2008); Prochnow et al. (Nature 445:447-451, 2007)]. Additionally, the structure of Apo3G-CD2 has also been determined using NMR [Chen et al. (Nature 452:116-119, 2008); Furukawa et al. (EMBO J 28:440-451, 2009); Harjes et al. (J Mol Biol, 2009)]. A detailed structural analysis of the APOBEC proteins and a comparison to other zinc-coordinating deaminases can facilitate our understanding of how APOBEC proteins bind nucleic acids, recognize substrates, and form oligomers. Here, we review the recent development of structural and functional studies that apply to Apo3G as well as the APOBEC deaminase family. PMID- 19547915 TI - Faithful after break-up: suppression of chromosomal translocations. AB - Chromosome integrity in response to chemically or radiation-induced chromosome breaks and the perturbation of ongoing replication forks relies on multiple DNA repair mechanisms. However, repair of these lesions may lead to unwanted chromosome rearrangement if not properly executed or regulated. As these types of chromosomal alterations threaten the cell's and the organism's very own survival, multiple systems are developed to avoid or at least limit break-induced chromosomal rearrangements. In this review, we highlight cellular strategies for repressing DNA break-induced chromosomal translocations in multiple model systems including yeast, mouse, and human. These pathways select proper homologous templates or broken DNA ends for the faithful repair of DNA breaks to avoid undesirable chromosomal translocations. PMID- 19547917 TI - H89, an inhibitor of PKA and MSK, inhibits cyclic-AMP response element binding protein-mediated MAPK phosphatase-1 induction by lipopolysaccharide. AB - OBJECTIVE: Lipopolysaccharide (LPS) stimulates the production of inflammatory cytokines and the amplification of immune responses via MAPK pathways. MAPK phosphatases (MKPs) feedback-regulate the activities of MAPKs to prevent excessive immunological functions. H89 has been used as an inhibitor of the protein kinase A (PKA) and mitogen- and stress-activated protein kinase (MSK) pathways. In view of the potential roles of PKA and MSK for MKP-1 induction and the ability of H89 to inhibit these kinases, this study examined the effect of H89 on MKP-1 induction by LPS and the role of cyclic-AMP response element binding protein (CREB) in the MKP-1 induction. RESULTS: H89 treatment inhibited increases in MKP-1 protein and mRNA levels, and gene transcription by LPS in Raw264.7 cells. Immunoblot, gel-shift, and chromatin-immunoprecipitation assays showed the activation of CREB by LPS, and the ability of H89 to inhibit it, suggesting that H89's inhibition of CREB may affect MKP-1 induction. In addition, H89 prevented the ability of LPS to induce other MKP genes (Dusp-2, 4, 8, and 16). Experiments using MAPK inhibitors showed that MAPKs are involved in CREB phosphorylation and MKP-1 induction, suggesting that CREB-mediated MKP-1 induction serves in part as a feedback-inhibitory loop of MAPKs. CONCLUSION: Our results demonstrate that H89 inhibits the activation of CREB and the CREB-mediated MKP-1 induction by LPS, which may result from its inhibition of PKA and MSK. PMID- 19547916 TI - Diversity, regulation, and genetic manipulation of plant mono- and sesquiterpenoid biosynthesis. AB - Among plant secondary metabolites, terpenoids are the most abundant and structurally diverse group. In addition to their important roles in pollinator attraction and direct and indirect plant defense, terpenoids are also commercially valuable due to their broad applications in the cosmetic, food, and pharmaceutical industries. Because of their functional versatility and wide distribution, great efforts have been made to decipher terpenoid biosynthetic pathways, to investigate the molecular mechanism determining their structural diversity, and to understand their biosynthetic regulation. Recent progress on the manipulation of terpenoid production in transgenic plants not only holds considerable promise for improving various plant traits and crop protection but also increases our understanding of the significance of terpenoid metabolites in mediating plant-environment interactions. PMID- 19547918 TI - Yttrium-90-ibritumomab tiuxetan in combination with intravenous busulfan, cyclophosphamide, and etoposide followed by autologous stem cell transplantation in patients with relapsed or refractory B-cell non-Hodgkin's lymphoma. AB - BACKGROUND: Radiolabelled immunotherapy agents have an increasingly significant role in autologous stem cell transplantation (ASCT) by improving the tolerability and increasing the efficacy of the conditioning regimen, thereby reducing the relapse risk. We evaluated the efficacy and safety of yttrium-90-ibritumomab tiuxetan ((90)Y-ibritumomab) combined with intravenous busulfan, cyclophosphamide, and etoposide (Bu/Cy/E) followed by ASCT in patients with relapsed or refractory B-cell non-Hodgkin's lymphoma (NHL). METHODS: Each patient received a single dose of (90)Y-ibritumomab (0.4 mCi/kg on day -14) with Bu/Cy/E as a conditioning regimen. RESULTS: The patient cohort consisted of 19 individuals (ten males), of median age 51 years (range, 25-63 years). Sixteen patients had received two or more chemotherapy regimens before transplantation. Histologies were diffuse large B-cell (n = 14), follicular (n = 2), mantle cell (n = 2), and Burkitt lymphoma (n = 1). All patients engrafted. The median time to neutrophil engraftment was 10 days and time to platelet engraftment was 10 days. Nineteen patients were evaluable for response. The objective overall response rate was 84.2% (16/19): continued CR, 36.8% (7/19); induced CR, 36.8% (7/19); and PR, 10.5% (2/19). With a median follow-up of 29.4 months (13.4-36.6), the estimated 3-year overall survival and event-free survival rates were 52.6% (95% confidence interval [CI] 45.8-59.4) and 26.3% (95% CI 19.8-32.8), respectively. Adverse events were similar to those seen historically with Bu/Cy/E alone, and there were no treatment related deaths. CONCLUSION: In conclusion, (90)Y ibritumomab with Bu/Cy/E and ASCT is feasible in patients with relapsed or refractory B-cell NHL, without increased toxicity. PMID- 19547919 TI - Phase II trial of sunitinib in patients with relapsed or refractory germ cell tumors. AB - Vascular endothelial growth factor (VEGF) overexpression and increased angiogenesis have been proposed as having biologic importance in germ cell tumors (GCT). We conducted a single-institution phase II trial of sunitinib, an oral inhibitor of the VEGF receptor, in patients with relapsed or refractory GCT. A Simon's two-stage design was used to determine the number of patients for enrollment. Responses were assessed using a modified version of Response Evaluation Criteria in Solid Tumors (RECIST), taking into account tumor marker changes. Dose modifications were made according to a nomogram for adverse events. Ten patients were enrolled. The first five received sunitinib 50 mg for four consecutive weeks, followed by a two-week break (4/2). Since four of five treated on this schedule had some tumor marker decline during the four-week "on" period, with subsequent rise during the two-week break, the dose was changed to 37.5 mg continuously for patients six to ten. However, only marker stabilization (no declines) was seen. Overall, there were no objective responses: Five had stable disease and five progressive disease (PD). Sunitinib was well tolerated; only one patient required a dose reduction due to grade 3 mucositis. Two patients experienced tumor-related hemorrhage (grade 3 and grade 1). All patients developed PD within three cycles. Sunitinib is well tolerated, but at standard doses, does not demonstrate significant activity in highly refractory GCT. Correlation between sunitinib treatment and tumor marker changes on the 50 mg 4/2 schedule suggest some pathways targeted by sunitinib (ie, angiogenesis) may be important to GCT biology. PMID- 19547920 TI - Sunitinib inducing tumor lysis syndrome in a patient treated for renal carcinoma. AB - Sunitinib is an oral antityrosine kinase inhibitor that has antiangiogenic and antitumor activities. It has been approved for the treatment of advanced RCC and for imatinib-refractory gastrointestinal stromal tumors (GIST). Tumor lysis syndrom can occur in solid tumors. We report a case of patient with metastatic RCC treated with sunitinib with a diagnosis of tumor lysis syndrome. PMID- 19547922 TI - Construction of intragenic synthetic riboswitches for detection of a small molecule. AB - Bacterial sensors, based on ligand-mediated genetic control systems, are promising for on-site chemical detection because sensing targets and generating signals do not require costly instrumentation. Here, we have constructed intragenic synthetic riboswitches without relying on high-throughput screening and demonstrated that the riboswitches can be harnessed to develop bacterial sensors displaying readily visible reporter signals in response to theophylline. In vivo imaging using the riboswitch showed target-specific changes in the expression of a green fluorescence protein reporter, which was visible even to the naked eye. PMID- 19547923 TI - Transaminase-catalyzed asymmetric synthesis of L-2-aminobutyric acid from achiral reactants. AB - Asymmetric synthesis of an unnatural amino acid was demonstrated by omega transaminase from Vibrio fluvialis JS17. L-2-Aminobutyric acid was synthesized from 2-oxobutyric acid and benzylamine with an enantiomeric excess higher than 99%. The reaction showed severe product inhibition by benzaldehyde, which was overcome by employing a biphasic reaction system to remove the inhibitory product from the aqueous phase. In a typical biphasic reaction (50 mM 2-oxobutyric acid, 70 mM benzylamine and 2.64 U/ml purified enzyme) using hexane as an extractant, conversion of 2-oxobutyric acid reached 96% in 5 h whereas only 39% conversion was obtained without the product extraction. PMID- 19547924 TI - The 3'-untranslated region of rice glutelin GluB-1 affects accumulation of heterologous protein in transgenic rice. AB - We compared the effect of the rice storage protein glutelin B-1 (GluB-1) terminator with the nopaline synthase (Nos) terminator on the accumulation of the modified house dust mite allergen mDer f 2 driven by the maize ubiquitin promoter in transgenic rice. Accumulation of mDer f 2 in transgenic seed and leaf using the GluB-1 terminator was greater than when using the Nos terminator construct. The mDer f 2 mRNA containing the GluB-1 3'UTR was processed and polyadenylated at the same sites as the native GluB-1 mRNA in the seeds but diverged in leaves of the transgenic plants. In contrast, the poly(A) sites of mDer f 2 containing Nos 3'UTR were more divergent in both seed and leaf. These results suggest that GluB 1 3'UTR functions as a faithful terminator and that termination at the specific sites may play an important role in mRNA stability and/or translatability, resulting in higher levels of protein accumulation. PMID- 19547925 TI - Comparative homology modeling-inspired protein engineering for improvement of catalytic activity of Mugil cephalus epoxide hydrolase. AB - The epoxide hydrolase (EH) of a marine fish, Mugil cephalus, was engineered to improve the catalytic activity based on comparative homology modeling. The 3-D crystal structure of the EH from Aspergillus niger was used as a template. A triple point mutant, F193Y for spatial orientation of the nucleophile (D199), W200L for removing electron density overlap between W200 and Y348, and E378D for good charge relay in the active site, was developed. The initial hydrolysis rate, the reaction time to reach 98 %ee, and yield were enhanced up to 35-fold, 26-fold and 32%, respectively, by homology modeling-inspired site-directed mutagenesis of M. cephalus EH. PMID- 19547927 TI - Cationic polyacrylamides enhance rates of starch and cellulose saccharification. AB - Adding a cationic polyacrylamide (c-PAM) to either the amylase mediated hydrolysis of corn starch or the hydrolysis of wood fiber by cellulase can enhance the initial hydrolysis rates, although a rate decrease can occur under some conditions. Several c-PAMs can serve as catalysts and the same c-PAM can improve the efficiency of both amylase and cellulase. The initial amylase rate approximately doubles; the analogous cellulase hydrolysis rate increases by about 40%. c-PAMs increase the binding of enzyme to substrate. PMID- 19547926 TI - Prokaryotic expression, purification and functional characterization of human FHL3. AB - Four and a half LIM domain protein 3 (FHL3) is a member of the family of LIM proteins and is involved in myogenesis, cytoskeleton reconstruction, cell growth and differentiation. The full-length FHL3 cDNA was cloned from human spleen cDNA library and inserted in a prokaryotic expression vector pBV220 and then the recombinant plasmid was transformed into E. coli JM109. The expression of the recombinant protein was induced at 42 degrees C. SDS-PAGE analysis showed that recombinant human FHL3 (rhFHL3) was mainly expressed as an inclusion body. After purification by HisTrap FF crude, the rhFHL3 was renatured by dialysis against renaturing buffer and identified by Western blot analysis using human FHL3 polyclonal antibody. The MTT assay showed that the purified rhFHL3 could inhibit HepG2 cell growth but promote the proliferation of ECV304 cells. In addition, the expression of angiogenin (Ang) gene was increased when ECV304 cells were pretreated with rhFHL3. PMID- 19547928 TI - Relationship between tissue plasminogen activator production and specific growth rate in Chinese hamster ovary cells cultured in mannose at low temperature. AB - Chinese hamster ovary (CHO) cells, producing human recombinant tissue plasminogen activator (tPA), were grown with mannose (5, 20 and 40 mM) instead of glucose at 31, 33 and 37 degrees C. The highest tPA concentration (1.5 mg l(-1) at 144 h of cultivation) and tPA specific production rate (47 ng 10(-6) cell h(-1)) were obtained at 31 degrees C and 40 mM mannose. Regardless of the temperature or mannose concentration used, an inverse relationship between the specific growth rate and tPA specific production rate was observed, suggesting that tPA production rate would be directly controlled by the growth rate. PMID- 19547929 TI - Improved detection of rhamnolipid production using agar plates containing methylene blue and cetyl trimethylammonium bromide. AB - Rhamnolipids, produced predominantly by Pseudomonas aeruginosa, are biosurfactants with important applications. For efficient culture screening according to rhamnolipid productivity, the method using agar plates containing methylene blue (MB) and cetyl trimethylammonium bromide (CTAB) was re-examined. An alternative set-up using a fixed underneath light source and image analysis software improved the detection of the circles formed due to complexation between anionic rhamnolipids and cationic MB/CTAB. The roles and effects of MB and CTAB concentrations and pH on the complexation phenomena are reported. PMID- 19547930 TI - Effects of variations in cadmium and lead levels in river sediments on local foods and body burden of local residents in non-polluted areas in Japan. AB - This study was initiated to examine if variations in the concentrations of cadmium (Cd) and lead (Pb) in water environment may affect metal levels in local foods and body burden of local residents in non-polluted areas in Japan. Two nationwide databases have been made available on concentrations of Cd in locally harvested brown rice and of Cd and Pb in sediments in local river beds. These data were combined with published data on metal concentrations in polished rice, food duplicates, and blood and urine from the residents. Cd in river sediments correlated significantly with those in brown rice, food duplicates, blood, and urine. Cd in food duplicates correlated with Cd in rice. In contrast, Pb concentrations in the river sediments either did not correlate or correlated only weakly with Pb in biological materials or food duplicates. Possible implication of the different behavior between Cd and Pb regarding the intensity of correlation was discussed with reference to the different routes of exposure to the elements. In conclusions, the Cd body burden on local residents in Japan is significantly influenced by Cd levels in water in the general environment, whereas water-borne Pb did not show clear correlation with the Pb body burden. PMID- 19547932 TI - The role of calcium, magnesium, and zinc in pre-eclampsia. AB - Pre-eclampsia is the most common medical complication of pregnancy associated with increased maternal and infant mortality and morbidity. Its exact etiology is not known, although several evidences indicate that various elements might play an important role in pre-eclampsia. This study was carried out to analyze and to compare the concentration of calcium, magnesium, and zinc in the serum of women with pre-eclampsia and in normal pregnant women. Fifty clinically diagnosed patients with pre-eclampsia (25 with mild and 25 with severe pre-eclampsia) and 50 normal pregnant controls were enrolled in this study. The serum calcium, magnesium, and zinc levels were estimated with an atomic absorption spectrophotometer. The mean serum levels of calcium, magnesium, and zinc in normal pregnant group were 2.45 +/- 0.18 mmol/L, 0.79 +/- 0.13 mmol/L, and 15.64 +/- 2.4 micromol/L, respectively, while in mild pre-eclamptic group, these were 2.12 +/- 0.15 mmol/L, 0.67 +/- 0.14 mmol/L, and 12.72 +/- 1.7 micromol/L, respectively. Serum levels in severe pre-eclamptic group were 1.94 +/- 0.09 mmol/L, 0.62 +/- 0.11 mmol/L, and 12.04 +/- 1.4 micromol/L, respectively. These results indicate that reduction in serum levels of calcium, magnesium, and zinc during pregnancy might be possible contributors in etiology of pre-eclampsia, and supplementation of these elements to diet may be of value to prevent pre eclampsia. PMID- 19547931 TI - Functional effects of short-term treatment with amiodarone on thyroid tissues of the rabbit. AB - The effect of short-term treatment with Amiodarone on thyroid gland tissue was studied in a group of 26 New Zealand albino rabbits. Ten rabbits were left untreated and served as controls; the remaining animals were treated with 10 mg/kg/day Amiodarone. The serum levels of serum triiodothyronine (T3), thyroxine (T4), and thyroid stimulating hormone (TSH) levels were measured at days 0 (baseline), 7, 30, and 45. The serum selenium levels were also measured, but only on days 0 and 45 of the experiment. At the end of the experiment the animals were sacrificed and the levels of selenium, T3, T4, and iodine were determined in thyroid tissue. After 30 days treatment the values of T3 were significantly lower than those of the untreated controls or the baseline levels (p < 0.001). The T4 level was significantly lower and the TSH value was significantly higher after 45 days of Amiodarone (p < 0.001). In thyroid tissue the T3, T4, and iodine levels were significantly higher in the treated group when compared to untreated controls (p < 0.05). These results show that Amiodarone induces changes in the hormone levels in both serum and thyroid tissues, as well as in the amount of iodine taken up by the thyroid gland in rabbits. PMID- 19547933 TI - [Neurological complication after a vertical infraclavicular brachial plexus block. Case report of possible differential diagnoses of a neurological deficit]. AB - A 72-year-old man with an obliteration of the brachial artery received a vertical infraclavicular block (VIP) for vascular surgery but 20 h after the operation a complete paresis of the affected extremity occurred. A new vascular obliteration could be excluded. During the diagnostic examination the patient noticed a snapping noise in the cervical column when moving his head and an abrupt recovery of the neurological deficits occurred. The radiological diagnostic provided no indication of cerebral ischemia or lesions of the brachial plexus. An additional diagnostic finding was a profound herniated vertebral disc with compression of the myelon. Fortunately, the neurological deficits completely returned to normal. PMID- 19547934 TI - [Brain death and intensive care medicine. Legal problems]. AB - "Death as the limit of the natural legal capacity is such a simple natural event that, just as for birth, it is not necessary to exactly determine its elements" declared the legal academic Friedrich Karl von Savigny 150 years ago. However, a stable legal order necessitates clear and if possible simple, catchy objective reference. The clinical concept of death at the point of irreversible cessation of circulation and breathing, in view of the possibilities of modern intensive medical care and the obvious redundant terms "brain death" and "cardiac circulation death", belongs to the past. Nowadays death is seen in the final cessation of all functions of the complete brain. Death as a legal term must, however, be "functionally" established with respect to the various aims and objectives of the Law. In this article the normative consequences of this distinction for intensive care medicine will be clarified. PMID- 19547935 TI - [Cricoid pressure. Protective manoeuvre or established nonsense?]. PMID- 19547936 TI - [Ultrasound guidance for placement control of central venous catheterization. Survey of 802 anesthesia departments for 2007 in Germany]. AB - OBJECTIVES AND METHODS: In 2007 a survey on the development of the current practice of using ultrasound to assist central venous catheter (CVC) placement was carried out in 802 departments of anesthesiology and intensive care medicine in hospitals with more than 200 beds in Germany. These data were compared to data from a survey in 2003. Additionally, data regarding control of CVC positioning were collected. RESULTS: The response rate was 58%. In these 468 departments approximately 340,000 CVCs are placed annually and 317 departments have access to an ultrasound machine. Ultrasound guidance is used by 188 (40%) departments for central venous cannulation. Of these only 24 (12.7%) use ultrasound routinely and 114 (60.6%) use it when faced with a difficult cannulation. Approximately one third of the users perform continuous ultrasound guidance for CVC placement. Equipment was not at disposal in 115 (41.1%) departments not using ultrasound for CVC placement did not possess the equipment and 93 (33.2%) did not consider ultrasound necessary. Positioning of CVCs was controlled either by electrocardiogram (ECG) guidance and/or chest radiograph in 92%. CONCLUSION: In Germany placement of central venous catheters is still usually based on anatomical landmarks. However, compared to 2003, ultrasound guidance for CVC placement is gradually being introduced (40% compared to 19%). Given the well documented advantages of ultrasound guidance compared to landmark based approaches for central venous cannulation, acquisition of this technology should belong to the training programme of an anesthesiologist. PMID- 19547937 TI - [Public health genomics. The future is built today!]. AB - The task of public health genomics (PHG) has become a challenge for all healthcare systems having major implications for future research and policy strategies. The various stakeholders in public health play a key role in translating the implications of genomics such as deriving from systems biology, epigenomics, integrative genomics or genome-environmental interactions. Recent advances in systems biology indicate that specific cellular functions are infrequently carried out by single genes, but rather by groups of cellular components. This network-based research is already starting to change nosology as well as to challenge population-based genetic screening or epidemiological methods like HTA. This knowledge will not only enable clinical interventions but also health promotion messages and disease prevention programs to be targeted at susceptible individuals as well as subgroups of the population (personalized healthcare). So far there has been no systematic integration of genome-based knowledge and technologies into public health research, policy, and practice. Thus, the public health agenda demands a vision that reaches beyond the research horizon to arrive at application and health impact of these innovations. The Public Health Genomics European Network (PHGEN) aims to fulfill this task in Europe. PMID- 19547938 TI - The impact of using mature compost on nitrous oxide emission and the denitrifier community in the cattle manure composting process. AB - The diversity and dynamics of the denitrifying genes (nirS, nirK, and nosZ) encoding nitrite reductase and nitrous oxide (N(2)O) reductase in the dairy cattle manure composting process were investigated. A mixture of dried grass with a cattle manure compost pile and a mature compost-added pile were used, and denaturing gradient gel electrophoresis was used for denitrifier community analysis. The diversity of nirK and nosZ genes significantly changed in the initial stage of composting. These variations might have been induced by the high temperature. The diversity of nirK was constant after the initial variation. On the other hand, the diversity of nosZ changed in the latter half of the process, a change which might have been induced by the accumulation of nitrate and nitrite. The nirS gene fragments could not be detected. The use of mature compost that contains nitrate and nitrite promoted the N(2)O emission and significantly affected the variation of nosZ diversity in the initial stage of composting, but did not affect the variation of nirK diversity. Many Pseudomonas-like nirK and nosZ gene fragments were detected in the stage in which N(2)O was actively emitted. PMID- 19547940 TI - Multilevel analysis of survey data. PMID- 19547941 TI - Droplet handling. AB - When quantitative analysis or quantitative chemical synthesis is performed using a micrototal analysis system (microTAS), the technologies for precise metering, transporting, and mixing of droplets are required. In this chapter, several technologies for the handling of droplets are described. For metering, dispensing and transporting of droplets, pneumatic and electrokinetic forces are used. Separation of cells and particles is also performed by electrical operation. Other handling technique, such as ultrasonic or centrifugal force applications, are also reviewed. Robotic synthesis devices or high throughput screening devices are promising applications for these technologies. PMID- 19547939 TI - Highly similar prokaryotic communities of sunken wood at shallow and deep-sea sites across the oceans. AB - With an increased appreciation of the frequency of their occurrence, large organic falls such as sunken wood and whale carcasses have become important to consider in the ecology of the oceans. Organic-rich deep-sea falls may play a major role in the dispersal and evolution of chemoautotrophic communities at the ocean floor, and chemosynthetic symbiotic, free-living, and attached microorganisms may drive the primary production at these communities. However, little is known about the microbiota thriving in and around organic falls. Our aim was to investigate and compare free-living and attached communities of bacteria and archaea from artificially immersed and naturally sunken wood logs with varying characteristics at several sites in the deep sea and in shallow water to address basic questions on the microbial ecology of sunken wood. Multivariate indirect ordination analyses of capillary electrophoresis single stranded conformation polymorphisms (CE-SSCP) fingerprinting profiles demonstrated high similarity of bacterial and archaeal assemblages present in timbers and logs situated at geographically distant sites and at different depths of immersion. This similarity implies that wood falls harbor a specialized microbiota as observed in other ecosystems when the same environmental conditions reoccur. Scanning and transmission electron microscopy observations combined with multivariate direct gradient analysis of Bacteria CE-SSCP profiles demonstrate that type of wood (hard vs. softwood), and time of immersion are important in structuring sunken wood bacterial communities. Archaeal populations were present only in samples with substantial signs of decay, which were also more similar in their bacterial assemblages, providing indirect evidence of temporal succession in the microbial communities that develop in and around wood falls. PMID- 19547944 TI - [Language development impairment and trisomy 8 mosaicism]. AB - Constitutional trisomy 8 mosaicism (46,XX/47,XX,+8 or 46,XY/47,XY,+8) is characterized by trisomic distribution of chromosomes in some but not all cells of the body. The full condition presents with physical stigmata, skeletal abnormalities and a mild to moderate cognitive impairment.Here we present a boy aged 3 years 10 months with partial trisomy 8 who was referred because of a language impairment. Because of known anomalies (corpus callous agenesis, deformities of the spine) a chromosomal analysis was initiated.This case highlights the necessity for an interdisciplinary diagnostic approach in children with language impairment and other constitutional disorders. PMID- 19547945 TI - Maternally inherited architecture in tertiary leaf beetles: paleoichnology of cryptocephaline fecal cases in Dominican and Baltic amber. AB - Complex ethological adaptations and intraspecific interactions leave few fossil traces. We document three Dominican (20 million years old [myo]) and Baltic (45 myo) amber fossils that exhibit firm evidence of highly integrated interactions between mothers and offspring in the diverse camptosomate lineage of beetles (Chrysomelidae, leaf beetles). As in contemporary species, these hard cases were initially constructed by mothers, then inherited and retained by offspring, which then elaborate this protective domicile with an unusual but economical building material, their feces. The three fossils are classified in the Subfamily Cryptocephalinae; two are classified in the tribe Chlamisini based on morphological evidence-the flattened head lacking a sharp keel and long legs with simple recurved untoothed claws. These diagnostic features are not clearly visible in the third specimen to permit more refined identification. These fossils provide more precise paleontological dating of tribal nodes within the cryptocephaline radiation of leaf beetles. These fossils are the first and earliest evidence of mother-offspring interaction, building behavior, and fecal recycling in Camptosomata beetles and of inheritance of architectural structures in beetles. PMID- 19547947 TI - [Psychopathological aspects of "raptus melancholicus"]. AB - The term "raptus melancholicus" is rarely used in clinical or scientific discussions. There is a lack of data on frequency and attribution to psychiatric disorders. It is found more often in the older literature where authors write of detailed personal experiences with this malicious phenomenon, but the published information is isolated as well. Clinicians and forensic psychiatrists judge raptus melancholicus to represent a risk for suicidal attempts as it appears suddenly and unexpectedly, arising from a state of delayed and inhibited psychomotor motility. This enigma, which sounds quite paradoxical, requires a psychopathological explanation as offered here. The inclusion of forensic aspects suggests that raptus melancholicus is not such a rare entity. PMID- 19547946 TI - [Informant report of cognitive changes in the elderly. A first evaluation of the German version of the IQCODE]. AB - BACKGROUND: Cognitive deficits occurring with dementia are frequently not reported by the affected subject. Therefore, informant reports from close relatives are especially important for the early diagnosis of dementia. Internationally, the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE) has been evaluated with positive results and is a widely used informant rated instrument for the diagnosis of cognitive decline. For the German speaking countries, norms and evaluation of the psychometric properties of the instrument are lacking. METHODS: Norms for the German long version of the IQCODE were established with 46 healthy elderly married couples. These were compared with respect to their concurrent and discriminative validity with groups of patients suffering from mild cognitive impairment (MCI, n=25), Alzheimer's or mixed dementia (AD, n=59) and frontotemporal lobe degeneration (FTLD, n=15). RESULTS: The German version of the IQCODE exhibited good psychometric properties and was able to best discriminate between cognitively intact and demented subjects with AD. Receiver-operating characteristic analyses indicated a cut-off score of 3.38 which corresponds well with the value given in international literature. Patients with MCI and with FTLD were also reliably distinguished from cognitively intact subjects. However, the instrument did not distinguish AD from FTLD with any significant degree of confidence. DISCUSSION: The German version of the IQCODE reliably discriminates cognitively intact persons from those suffering from MCI or cortical dementia, but not between different types of cortical dementia, such as AD and FTLD. The IQCODE is an efficient informant-rated screening instrument for the early diagnosis of cognitive decline and dementia. PMID- 19547948 TI - [Cryoablation of localized prostate cancer. Current state]. AB - Due to continuous technical developments for more than half a century followed by better clinical results with minimal side effects, cryoablation of the prostate for localized prostate cancer has evolved as a true alternative therapeutic option in selected cases. The current version of cryoablation has almost nothing in common with those versions established in the 1960s and 1970s and further developed in the 1980s and 1990s. The present version is minimally invasive and has a high efficacy for treatment of high risk carcinomas and failure of other therapeutic modalities. Cryoablation of the prostate is indicated if there are absolute or relative contraindications for radical surgery. In salvage cases cryoablation is the therapy of choice for localized prostate cancer. Standardization of the procedure, definition of freeze-thaw cycles and structured training programs have led to this status. PMID- 19547949 TI - Height growth velocity, islet autoimmunity and type 1 diabetes development: the Diabetes Autoimmunity Study in the Young. AB - AIMS/HYPOTHESIS: Larger childhood body size and rapid growth have been associated with increased type 1 diabetes risk. We analysed height, weight, BMI and velocities of growth in height, weight and BMI, for association with development of islet autoimmunity (IA) and type 1 diabetes. METHODS: Since 1993, the Diabetes Autoimmunity Study in the Young (DAISY) has followed children at increased type 1 diabetes risk, based on HLA-DR, -DQ genotype or family history, for the development of IA and type 1 diabetes. IA was defined as the presence of autoantibodies to insulin, GAD or protein tyrosine phosphatase islet antigen 2 twice in succession, or autoantibody-positive on one visit and diabetic at the next consecutive visit within 1 year. Type 1 diabetes was diagnosed by a physician. Height and weight were collected starting at age 2 years. Of 1,714 DAISY children <11.5 years of age, 143 developed IA and 21 progressed to type 1 diabetes. We conducted Cox proportional hazards analysis to explore growth velocities and size measures for association with IA and type 1 diabetes development. RESULTS: Greater height growth velocity was associated with IA development (HR 1.63, 95% CI 1.31-2.05) and type 1 diabetes development (HR 3.34, 95% CI 1.73-6.42) for a 1 SD difference in velocity. CONCLUSIONS/INTERPRETATION: Our study suggests that greater height growth velocity may be involved in the progression from genetic susceptibility to autoimmunity and then to type 1 diabetes in pre-pubertal children. PMID- 19547950 TI - Novel pharmacological approaches to combat obesity and insulin resistance: targeting skeletal muscle with 'exercise mimetics'. AB - Chronic diseases arising from obesity will continue to escalate over coming decades. Current approaches to combating obesity include lifestyle measures, surgical interventions and drugs that target weight reduction or the metabolic consequences of obesity. Lifestyle measures including physical activity are usually the primary strategy, but these are of limited long-term efficacy because of failure to maintain behavioural change. An alternative approach used to elicit the benefits of exercise training and overcome the problems of long-term compliance is to develop drugs that mimic aspects of the trained state. Elucidation of metabolic pathways responsive to exercise in various tissues, particularly skeletal muscle, was an important antecedent to the promising concept of drugs that may mimic specific aspects of the exercise response. From an obesity perspective, an important aim is to develop an agent that reduces body fat and improves metabolic homeostasis. This review focuses on promising metabolic signalling pathways in skeletal muscle that may yield 'exercise mimetic' targets. PMID- 19547951 TI - [Ultrasound measurement of humeral torsion in children and adolescents with hemiplegic cerebral palsy]. AB - BACKGROUND: Torsional deformity in the lower extremities is well known in children and adolescents with hemiplegic cerebral palsy, however, little or nothing is known about possible differences in torsional behavior of the upper extremities. Alterations of torsion in the lower extremities influence the gait, alterations in the rotation behavior of the upper extremities, for example after fractures, lead to limited movement of the arms or to an altered movement radius. The aim of this study was to investigate whether there is a difference in humeral torsion between affected and non-affected arms in children and adolescents with hemiplegic cerebral palsy and which factors play a decisive role. PATIENTS AND METHODS: A total of 33 children and adolescents with hemiplegic cerebral palsy (mean age 9.7 years, range 3-16 years) answered a questionnaire and were examined clinically, photographically and with a new ultrasound method. Severity of cerebral palsy was classified into three levels. Ultrasound measurements of humeral torsion were compared with the results from a normal collective (n=149). RESULTS: In approximately half (n=16, 48.5%) of the 33 children investigated, humeral torsion was directed more internally, in 17 (51.5%) children humeral torsion was directed more externally compared to the healthy side. The difference in humeral torsion between the affected and non-affected sides was significant (p<0.05) in both groups. Differences in humeral torsion between the affected and non-affected sides in children with hemiplegic cerebral palsy were significantly increased (p<0.00001) compared to differences in humeral torsion between left and right arms in healthy children. A correlation with the level of severity of hemiplegic cerebral palsy was observed but there was no correlation with age. Children and adolescents with external torsional deformity had a smaller range of rotation, a larger muscle force deficit of the affected upper extremity and did less sport using the affected arm than children and adolescents with internal torsional deformity. CONCLUSION: Humeral torsion deformity in children and adolescents with hemiplegic cerebral palsy was proven to exist in addition to the already well known difference in torsion of the lower extremities. Analogous to increased activity of external or internal rotator muscles codetermined by the palsy, increased external or internal humeral torsion occurs. External torsion deformity was associated with a smaller range of motion and seemed to have a greater importance for routine daily activities (e.g. muscle force, sports ability) than internal torsion deformity. PMID- 19547952 TI - Noninvasive pressure-support ventilation in immunocompromised children with ARDS: a feasibility study. AB - OBJECTIVE: To verify the feasibility of non-invasive ventilation (NIV) in immunocompromised children affected by ARDS. SETTING: University Hospital PICU. PATIENTS: Twenty-three consecutive immunocompromised children treated with NIV for ARDS. INTERVENTIONS: All consecutive patients received NIV through a face mask or a helmet. RESULTS: No differences were found regarding admission data and severity scores between NIV responders and non-responders. Early and sustained improvement in PaO2/FiO2 ratio were observed in 82 and 74% of cases, respectively. 13 out of 23 patients (54.5%) avoided intubation and were discharged from the PICU; ten patients required intubation: two of them survived and eight patients died (two refractory hypoxemia, three septic shock, three multi-organ failure). PICU and intra-hospital mortality was significantly higher for NIV-nonresponders (P < 0.001). PICU stay was significantly shorter for NIV responders (P = 0.03). NIV responders had significantly lower heart and respiratory rate at the end of treatment (P < 0.001 and P = 0.048, respectively). CONCLUSIONS: NIV administration is feasible and well tolerated in immunocompromised children with ARDS. A short NIV trial can be used to verify the usefulness of the technique. A randomized controlled trial is needed to confirm the efficacy of NIV in immunocompromised children requiring ventilatory support for ARDS. PMID- 19547953 TI - Factor replacement for Australian snakebite coagulopathy: a re-evaluation? PMID- 19547954 TI - Clotting factor replacement and recovery from snake venom-induced consumptive coagulopathy. AB - INTRODUCTION: Using clotting factors (fresh frozen plasma and/or cryoprecipitate) to treat snake venom-induced consumptive coagulopathy (VICC) is controversial. We aimed to determine if factor replacement after antivenom is associated with an earlier return of coagulation function. METHODS: We retrospectively analysed VICC cases due to brown snake (genus Pseudonaja), tiger snake (Notechis, Tropidechis, and Hoplocephalus), and taipan (Oxyuranus) envenoming. Recovery of international normalized ratio (INR)/prothrombin time (PT) was compared between patients who did not receive factor replacement and those who did, and between patients who received factor replacement or=4 packed red cells (PRCs) and a worse clinical outcome and higher mortality. The associated mortality rate was higher for patients who underwent bypass surgery when they had Hb 8.9 g/dL and were transfused >or=4 PRCs. CONCLUSIONS: Low haemoglobin concentrations and transfusions in patients undergoing cardiac surgery are associated with increased morbidity and mortality. Also, anemia and transfusions are associated with poor outcome. Therefore, intra- and postoperative bleeding seem to be a risk factor in patients undergoing cardiac surgery. PMID- 19547958 TI - Predicting drug metabolism-dependent toxicity. PMID- 19547959 TI - Determinants for the cAMP-binding site at the S-adenosylhomocysteine-hydrolase. AB - S-Adenosylhomocysteine-hydrolase (AdoHcy-hydrolase) catalyzes the reversible hydrolysis of S-adenosylhomocysteine (AdoHcy) to adenosine (Ado) and homocysteine (Hcy). Since Ado competes with cAMP at the high affinity-binding site of the enzyme, we determined the effect of cAMP on enzyme activity and its binding characteristics to purified AdoHcy-hydrolase from bovine kidney in its native, in its fully oxidized (NAD(+)), and in its fully reduced (NADH) form. cAMP (10 micromol/l) enhanced the hydrolytic activity of native AdoHcy-hydrolase by 35%, whereas the activity of the enzyme in its NAD(+) form was not stimulated by cAMP. In contrast to azido-Ado, binding of azido-cAMP did not inhibit the enzymatic activity of AdoHcy-hydrolase. Furthermore, cAMP did not prevent the Ado induced inhibition of the AdoHcy hydrolysis. Saturation binding experiments with the three different forms of AdoHcy-hydrolase, native, NAD(+), and NADH showed only one binding site with high affinity. This binding site was identified after photoaffinity labeling of the enzyme with 8-azido-[2-(3)H]-cAMP. One photolabeled peptide was isolated as Trp(310)-Val(325) from each AdoHcy-hydrolase from native, NAD(+), and NADH. The cAMP-labeled peptide is located in the NAD-binding domain of AdoHcy-hydrolase. In conclusion, our data show that the cAMP-binding site at the AdoHcy-hydrolase is independent of the NAD(+)/NADH ratio of the enzyme and is identical with the high affinity-binding site of Ado. Moreover, cAMP did not interact with the catalytic site of AdoHcy-hydrolase and did not act as an allosteric effector for the AdoHcy-hydrolase. PMID- 19547961 TI - Ultrasonography and clinical findings in children with epididymitis, with and without associated lower urinary tract abnormalities. AB - BACKGROUND: Epididymitis is most commonly idiopathic but can also be associated with urinary tract abnormalities (UTAs). The distinctive clinical and imaging findings of children with epididymitis and underlying UTAs are not known. OBJECTIVE: To describe clinical and imaging findings in children with epididymitis and the association with UTAs. MATERIAL AND METHODS: The study group included all children evaluated for epididymitis confirmed by scrotal US in a 6 year period. The clinical and imaging findings and disease recurrence were compared between children with and without UTAs. RESULTS: A total of 47 boys (mean/SD 9.61/4.40 years, range 0.1 to 17.1 years) met the entry criteria, of whom 17 had UTAs. The most common UTAs were hypospadias, neurogenic bladder, and functional bladder abnormality (six each). Age at presentation and likelihood of testicular swelling or hydrocele was not different between children with and without UTAs. Marked epididymal swelling was more common in children with UTAs (9/17, 53%) than in those without UTAs (5/30, 17%; P=0.02), as was recurrent epididymitis (with UTAs, 9/17, 53%; without UTAs, 5/30, 17%; P=0.02). Chronic epididymitis (five children), presentation with scrotal mass (four), and epididymal abscess (two) occurred only in children with UTAs. CONCLUSION: Children with epididymitis who have UTAs are more likely to present with marked epididymal swelling, develop recurrent disease, and have a more protracted course. PMID- 19547957 TI - The educational environment for training in intensive care medicine: structures, processes, outcomes and challenges in the European region. AB - OBJECTIVE: To characterise the training environment in ICM across Europe, with a particular focus on factors influencing competency-based training. METHOD: A cross-sectional web-based survey completed by the national coordinator for the CoBaTrICE (Competency-Based Training in Intensive Care medicinE) programme in each of 28 European countries. RESULTS: Since the last survey in 2004, 50% of EU countries have modified their training programmes. Seven have already adopted the CoBaTrICE programme since its completion in 2006. Multidisciplinary access to ICM training ('supraspeciality' model) is available in 57%, most commonly as a 2-year training programme. National examinations are held by 26 (93%); in 24 (86%) this is a mandatory exit exam; ten use the European Diploma of Intensive Care (EDIC). A formal national system for quality assurance of ICM training exists in only 18 (64%) countries. National standards for approving hospitals as training centres vary widely. In 29% there is no designated specialist with responsibility for training at the local level. Time for teaching was cited as inadequate by 93% of respondents; only 21% of trainers receive contractual recognition for their work. In 39% there is no protected teaching time for trainees. Half of countries surveyed have no formal system for workplace-based assessment of competence of trainees. CONCLUSIONS: There is considerable diversity in pedagogic structures, processes and quality assurance of ICM across Europe. National training organisations should develop common standards for quality assurance, health systems need to invest in educator support, and the EU should facilitate harmonisation by recognising ICM as a multidisciplinary speciality. PMID- 19547962 TI - Fulminant limb and retroperitoneal necrotizing fasciitis in a 15-year-old girl with Fanconi anaemia. AB - Necrotizing fasciitis (NF) is an uncommon soft-tissue infection in children that carries a high mortality rate. We present a 15-year-old girl with chronic pancytopenia secondary to Fanconi anaemia who developed extensive NF of the lower limb, which unfortunately resulted in a fatal outcome. Immunodeficiency is a known risk factor for the development of this condition. The findings in this case demonstrate that patients with Fanconi anaemia may be susceptible to NF and that the clinical course may be more aggressive due to underlying immunosuppression. Prompt diagnosis of NF is vital in order to initiate appropriate treatment and to optimize patient outcome. Radiological investigation demonstrated extensive soft-tissue gas and destruction affecting the entire lower limb, abdominal wall and retroperitoneum, which led to timely definitive diagnosis and management. PMID- 19547963 TI - Tracheal compression due to an elongated aortic arch in patients with congenital heart disease: evaluation using multidetector-row CT. AB - BACKGROUND: The airway can become obstructed as a result of compression by an elongated aortic arch. OBJECTIVE: In this study we evaluated tracheal compression using multidetector-row CT in patients with congenital heart disease and an elongated aortic arch. MATERIALS AND METHODS: The trachea was measured at the level of the aortic arch in 205 children and young adults and then the severity of tracheal compression was determined by measuring the tracheal diameter ratio (short axis diameter/long axis diameter). Patients were divided as follows: group I (normal aortic arch; n=166), group II (transversely running aortic arch; n=22), and group III (elongated aortic arch; n=17). From the viewpoint of the relationship of the great arteries, group II had D-malposition, and group III had L-malposition. RESULTS: Age, height, weight and body surface area were significantly correlated with the short and long axis diameter in group I. There was a negative correlation between tracheal diameter ratio and the physical size parameters. The tracheal diameter ratio in group III was 0.50+/-0.13, which was significantly lower than in groups I and II (P<0.01 and 0.05, respectively). CONCLUSION: Even apparently asymptomatic patients with an elongated aortic arch can have tracheal compression. An elongated aortic arch may be a useful predictor of tracheal compression. PMID- 19547964 TI - Paratesticular rhabdomyosarcoma presenting as thickening of the tunica vaginalis. AB - Pediatric paratesticular primary malignant tumors are rare, and most of them correspond to rhabdomyosarcomas (RMS). Paratesticular RMS has a better prognosis than RMS originating in other locations, because of its favorable histology and possible early diagnosis. The diagnosis relies upon the US findings of a paratesticular hypervascular mass in an adequate clinical setting. We report the unusual appearance of an infiltrating RMS tumor in a 3-year-old boy. The tumor presented as thickening of the tunica vaginalis. Atypical US appearance and discordance with clinical features caused suspicion of a tumoral process, allowing early diagnosis and treatment. PMID- 19547965 TI - Pediatric radiology in the Philadelphia region: a history of pedigrees and legacies. AB - The specialty of pediatric radiology in the Philadelphia region has grown and evolved over the past eight decades originating from early "visiting" radiologists to Drs. Hope and Kirkpatrick, the "giants" of the 1950s and 1960s, to over 40 actively practicing pediatric radiologists. Clinical excellence, commitment to teaching, and advancement of knowledge through research remain the goals and ideals, much as they were many years ago. Philadelphia has been a fertile home and environment for this development, mostly because of outstanding leaders and role models who have trained and influenced generations of pediatric radiologists. The purpose of this article is to tell the story of this growth and development and to explore the intellectual origins, professional "genealogy," and legacies left by those who created this tradition. PMID- 19547960 TI - Are adolescents more vulnerable to drug addiction than adults? Evidence from animal models. AB - BACKGROUND AND RATIONALE: Epidemiological evidence suggests that people who begin experimenting with drugs of abuse during early adolescence are more likely to develop substance use disorders (SUDs), but this correlation does not guarantee causation. Animal models, in which age of onset can be tightly controlled, offer a platform for testing causality. Many animal models address drug effects that might promote or discourage drug intake and drug-induced neuroplasticity. METHODS: We have reviewed the preclinical literature to investigate whether adolescent rodents are differentially sensitive to rewarding, reinforcing, aversive, locomotor, and withdrawal-induced effects of drugs of abuse. RESULTS AND CONCLUSIONS: The rodent model literature consistently suggests that the balance of rewarding and aversive effects of drugs of abuse is tipped toward reward in adolescence. However, increased reward does not consistently lead to increased voluntary intake: age effects on voluntary intake are drug and method specific. On the other hand, adolescents are consistently less sensitive to withdrawal effects, which could protect against compulsive drug seeking. Studies examining neuronal function have revealed several age-related effects but have yet to link these effects to vulnerability to SUDs. Taken together, the findings suggest factors which may promote recreational drug use in adolescents, but evidence relating to pathological drug-seeking behavior is lacking. A call is made for future studies to address this gap using behavioral models of pathological drug seeking and for neurobiologic studies to more directly link age effects to SUD vulnerability. PMID- 19547967 TI - Enhancing pozzolana colonization by As(III)-oxidizing bacteria for bioremediation purposes. AB - The colonization of pozzolana by an As(III)-oxidizing bacterial consortium was monitored from the first hours of bacterial adhesion to 6 weeks of development under fed-batch conditions, using adapted ultrasonic dislodging and crystal violet staining procedures to determine the biofilm adhering to the complex surfaces. The effect of temperature, arsenic concentration, and presence or absence of yeast extract (YE) on the amount of biofilm biomass and on the As(III) oxidation were assessed to test the biofilm's resilience and optimize the colonization. Fed-batch cultures allow twice as much pozzolana colonization as that obtained under batch conditions. In addition, As(III) oxidation and the quantities of biomass under fed-batch culture conditions were the same at 14 degrees C and 25 degrees C. Whereas YE improves (+150%) bacterial adhesion during the first 2 h, its impact in the longer term appears to be less significant biofilm formation in presence of YE after 5 weeks was no greater than biofilm formation in the absence of YE. Finally, YE involves a drastic (-70%) decrease of As(III) oxidation. Preliminary tests for drinking-water bioremediation revealed the ability of Cheni Arsenic Oxidizing 1 biofilms to remain and retain As(III) oxidation activity at low As(III) concentrations (50 microg l(-1)). PMID- 19547966 TI - Soluble cytoplasmic expression, rapid purification, and characterization of cyanovirin-N as a His-SUMO fusion. AB - Cyanovirin-N (CVN) is a promising antiviral candidate that has an extremely low sequence homology with any other known proteins. The efficient and soluble expression of biologically functional recombinant CVN (rCVN) is still an obstacle due to insufficient yield, aggregation, and abnormal modification. Here, we describe an improved approach to preparing native rCVN from Escherichia coli more efficiently. A fusion gene consisting of cvn and sumo (small ubiquitin-related modifier) and a hexahistidine tag was constructed according to the codon bias of the host cell. This small ubiquitin-related modifier (SUMO)-fused CVN is expressed in the cytoplasm of E. coli in a folded and soluble form (>30% of the total soluble protein), yielding 3 to 4 mg of native rCVN from 1 g of wet cells to a purity up to 97.6%. Matrix-assisted laser desorption ionization coupled to time-of-flight mass spectrometry and reverse-phase high-performance liquid chromatographic analysis showed that the purified rCVN was an intact and homogeneous protein with a molecular weight of 11,016.68 Da. Potent antiviral activity of rCVN against herpes simplex virus type 1 and human immunodeficiency virus type 1/IIIB was confirmed in a dose-dependent manner at nanomolar concentrations. Thus, the His-SUMO double-fused CVN provides an efficient approach for the soluble expression of rCVN in the cytoplasm of E. coli, allowing an alternative system to develop bioprocess for the large-scale production of this antiviral candidate. PMID- 19547968 TI - Isolation of a thermostable legume chitinase and study on the antifungal activity. AB - Chitinases are listed as one class of pathogenesis-related proteins, and they have become a popular research topic because of their resistance to plant pathogenic diseases. A chitinase with antifungal activity was isolated from the Canadian cranberry beans (Phaseolus vulgaris). The procedure included extraction, ammonium sulfate precipitation, affinity chromatography on Affi-gel blue gel, CM Sephadex C-50, and Sephadex G-75. There was an almost 108-fold increase in specific activity of the purified chitinase compared with that of the crude extract. The enzyme exhibited a molecular mass of 30.6 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis both under reducing and non-reducing conditions, indicating that it was a monomeric protein. The pI was determined to be 7.6 by isoelectric-focusing electrophoresis. The optimum pH and the optimum temperature for activity towards N-acetyl-glucosamine was 5.4 and 40-55 degrees C, respectively. It exerted a potent inhibitory action toward fungal species including Botrytis cinerea, Physalospora piricola, Fusarium oxysporum, and Pythium aphanidermatum. The chitinase was thermostable up to 58 degrees C in both enzymatic reaction and antifungal activity. The present findings demonstrated a thermostable chitinase from cranberry beans with potentially exploitable significance. PMID- 19547969 TI - Effects of pulse feeding of beet molasses on recombinant benzaldehyde lyase production by Escherichia coli BL21(DE3). AB - The effect of fed-batch operation (FBO) strategy was investigated using pretreated-beet molasses, containing galactose that induces the lac promoter, on benzaldehyde lyase (BAL) production by recombinant Escherichia coli BL21(DE3)pLySs. After batch cultivation with 30 g l(-1) pretreated-beet molasses consisting of 7.5 g l(-1) glucose and 7.5 g l(-1) fructose, three FBO strategies were applied at dissolved oxygen (=40%) cascade to air-flow rate. In FBO1 when air-flow rate decreased considerably, feed was given to the system in pulses in such a way that pretreated-beet molasses concentration increased by 10 kg m(-3) (containing 2.5 g l(-1) glucose+2.5 g l(-1) fructose); however, decrease in air flow rate demonstrated only the absence of glucose but not fructose. Thus, in FBO2 when fructose and glucose were completely utilized, pretreated-beet molasses was pulse-fed and its concentration increased by 10 g l(-1). In FBO3 with the decreased amount of pretreated-beet molasses (6 g l(-1)), shift response time from glucose to fructose consumption was avoided, and glucose and fructose consumptions were well correlated with air-flow rate, and the highest CX (8.04 g l(-1)) and BAL (2,315 U ml(-1)) production were obtained (t=24 h) with the highest substrate yield on cell and product formation. PMID- 19547970 TI - Type II autosomal dominant osteopetrosis: radiological features in two families containing five members with asymptomatic and uncomplicated disease. AB - In this study we analysed the imaging patterns in two families containing five members with asymptomatic and uncomplicated autosomal dominant osteopetrosis (ADO II), and we report new and uncommon radiological manifestations. These findings might be useful in the context of reducing the incidence of fractures and other orthopaedic complications. Diffuse pelvic sclerosis on radiographs was observed incidentally in two patients. Both cases were asymptomatic, and the patients had never suffered a fracture. The suggestion of ADO II was raised. A detailed medical history, an imaging survey, and a haematological study were obtained so that other rare causes of osteosclerosis could be ruled out. No genetic study was conducted. All their first-degree relatives were also examined. Bony sclerosis was observed in five patients, and the radiological findings were analysed. A not previously reported thickening of the skull base without cranial nerve palsy or optic nerve atrophy was revealed in all patients. Scoliosis was present in three of them. This has been reported previously only once in ADO II. No lower limb deformity was detected. This study provided information on the pattern of radiological features in familial asymptomatic ADO II. These data on new and rare imaging findings will increase the diagnostic awareness of physicians and will guide a thorough investigation of the entire family. This might result in a consequent decrease in the incidence of fractures and other orthopaedic complications. PMID- 19547971 TI - Commentary on: Validation of sentinel lymph node dissection in prostate cancer: experience in more than 2,000 patients. PMID- 19547972 TI - Which osteotomy for a valgus knee? AB - A valgus knee is a disabling condition that can affect patients of all ages. Antivalgus osteotomy of the knee is the treatment of choice to correct the valgus, to eliminate pain in the young or middle age patient, and to avoid or delay a total knee replacement. A distal femoral lateral opening wedge procedure appears to be one of the choices for medium or large corrections and is particularly easy and precise if compared to the medial femoral closing wedge osteotomy. However, if the deformity is minimal, a tibial medial closing wedge osteotomy can be done with a faster healing and a short recovery time. PMID- 19547974 TI - Gibberella zeae chitin synthase genes, GzCHS5 and GzCHS7, are required for hyphal growth, perithecia formation, and pathogenicity. AB - Gibberella zeae causes Fusarium head blight of cereal crops, and sexual spores of the fungus play an important role as primary inocula. We isolated a restriction enzyme-mediated integration (REMI) transformant, ZH431, of G. zeae with defects in perithecia formation and virulence. Integration of the REMI vector resulted in disruption of GzCHS7 gene, which encodes a putative class VII chitin synthase with high similarity to Fusarium oxysporum ChsVb. A second chitin synthase, GzCHS5, is adjacently located in a head-to-head configuration with GzCHS7, and its deduced protein sequence showed similarity with a class V chitin synthase in F. oxysporum. Neither DeltaGzChs5 nor DeltaGzChs7 mutants produced perithecia or caused disease on barley heads. Microscopic observation revealed that both mutants formed balloon-shaped hyphae and intrahyphal hyphae and that cell wall rigidity of the mutants was weaker than that of the wild-type strain. Transcription profiles of GzCHS5 and GzCHS7 were not altered in DeltaGzChs7 and DeltaGzChs5, respectively, suggesting that transcription regulations of the genes are independent of each other. Our results demonstrate that GzCHS5 and GzCHS7 are indispensable for perithecia formation and pathogenicity as well as normal septa formation and hyphal growth in G. zeae. PMID- 19547973 TI - Complications of closing wedge high tibial osteotomy. AB - Closing wedge high tibial osteotomy is a common, effective and well-established procedure to treat unicompartment osteoarthrosis of the knee. It is, however, not without its complications. This article will discuss some of these complications and present an overview of the current literature. It will examine current thoughts on aetiology, techniques to try to avoid, and methods of treatment of these complications. PMID- 19547975 TI - The MAPKK kinase ChSte11 regulates sexual/asexual development, melanization, pathogenicity, and adaptation to oxidative stress in Cochliobolus heterostrophus. AB - All fungi use multiple mitogen-activated protein kinase (MAPK) cascades to respond to external signals to regulate specialized responses. In this study, we cloned and characterized a putative MAPKKK gene ChSte11, orthologous to yeast STE11, of Cochliobolus heterostrophus. DeltaChste11 strains showed defects in conidiation, sexual development, melanization and the formation of appressoria. These mutants were significantly less virulent on corn plants than the wild type. Similar phenotypes were observed in mutants of Chk1-MAPK, a putative downstream protein kinase of ChSte11. These results suggested that ChSte11 regulates various morphological changes and pathogenicity via Chk1 MAPK. Both DeltaChste11 and Deltachk1 strains showed severe sensitivity to oxidative stress, hydrogen peroxide, and heavy metals, cupric or ferric cations. DeltaBmhog1 strains, mutants of the HOG1-type MAPK, did not show sensitivity to these forms of stress. Our results strongly suggested that the Ste11-type MAPKKK regulates not only various morphological changes and pathogenicity, but also adaptations to stress via Chk1-type MAPK in filamentous fungi. PMID- 19547976 TI - The positivity of rheumatoid factor and anti-cyclic citrullinated peptide antibody in nonarthritic patients with chronic hepatitis C infection. AB - Chronic hepatitis C virus (HCV) has extrahepatic autoimmune properties and a variety of autoantibodies were found in patients with HCV. Patients with HCV infection may have rheumatic symptoms and signs, and 50-70% of the cases may contain rheumatoid factor (RF). The increased prevalence of RF in patients with HCV infection diminishes the diagnostic specificity of serum RF for rheumatoid arthritis (RA) in patients with HCV. Therefore, the presence of RF mostly does not help in distinguishing between RA and HCV-associated rheumatic symptoms. In this study, we studied whether cyclic citrullinated peptide (CCP) antibody, a highly specific biomarker for RA in the general population, was useful for the diagnosis of RA in nonarthritic patients with HCV (hepatitis C virus). Blood samples from 39 patients with chronic HCV infection, 87 normal sera from volunteer blood donors and 108 blood samples from patients with rheumatoid arthritis, from the rheumatology clinic, were taken. RF was measured using the Dade-Behring nephelometer and antibodies to CCP were measured with ELISA. According to statistical analysis, the sensitivity, specificity and positive predictive value of the anti-CCP test was superior to the RF test. Cyclic citrullinated peptide antibody is a more useful test than RF among patients with chronic HCV infection without arthritis. PMID- 19547977 TI - A monoallelic double mutation as a cause for TNF receptor-associated periodic fever syndrome. AB - Hereditary periodic fever syndromes (HPFSs) are a subset of human autoinflammatory diseases characterized by periodic episodes of fever and signs of inflammation with or without involvement of inner organs. In this paper, we report phenotypic features of an index patient and affected family members that present a previously not described mutation type in the TNFRSF1A gene. The phenotype of a HPFS of affected family members was shown to be associated with two monoallelic mutations in TNFRSF1A. Primarily, the index patient was clinically diagnosed as being affected by familial Mediterranean fever (FMF). However, with molecular genetic analyses, it could be shown that the patient was in fact affected by tumor necrosis factor receptor-associated periodic syndrome, which requires a different therapy when compared with FMF. Thus, molecular genetic analyses of currently known disease loci enable the most precise diagnosis presently available and are consequently the basis for the most effective therapeutic intervention. PMID- 19547978 TI - Plasma from systemic lupus patients compromises cholesterol homeostasis: a potential mechanism linking autoimmunity to atherosclerotic cardiovascular disease. AB - Atherosclerotic cardiovascular disease (ASCVD) contributes to morbidity and mortality in systemic lupus erythematosus (SLE). Immunologic derangements may disrupt cholesterol balance in vessel wall monocytes/macrophages and endothelium. We determined whether lupus plasma impacts expression of cholesterol 27 hydroxylase, an anti-atherogenic cholesterol-degrading enzyme that promotes cellular cholesterol efflux, in THP-1 human monocytes and primary human aortic endothelial cells (HAEC). THP-1 monocytes and HAEC were incubated in medium containing SLE patient plasma or apparently healthy control human plasma (CHP). SLE plasma decreased 27-hydroxylase message in THP-1 monocytes by 47 +/- 8% (p < 0.008) and in HAEC by 51 +/- 5.5% (n = 5, p < 0.001). THP-1 macrophages were incubated in 25% lupus plasma or CHP and cholesterol-loaded (50 microg ml(-1) acetylated low density lipoprotein). Lupus plasma more than doubled macrophage foam cell transformation (74 +/- 3% vs. 35 +/- 3% for CHP, n = 3, p < 0.001). Impaired cholesterol homeostasis in SLE provides further evidence of immune involvement in atherogenesis. Strategies to inhibit or reverse arterial cholesterol accumulation may benefit SLE patients. PMID- 19547979 TI - The role of capillaroscopy in differentiation of primary and secondary Raynaud's phenomenon in rheumatic diseases: a review of the literature and two case reports. AB - The purpose of this study is to study and systematize the current knowledge about the role of capillaroscopy in differentiation of primary and secondary Raynaud's phenomenon (RP) in rheumatic diseases. This method is a review of the literature. Capillaroscopy is of crucial value for diagnosis and differentiation of primary and secondary RP in rheumatic diseases. The appearance of abnormal capillaroscopic pattern inherits high positive predictive value for the development of systemic rheumatic disease. The most specific pattern is found in systemic sclerosis (SSc), so called "scleroderma pattern", which is characterized by the presence of dilated capillaries, hemorrhages, avascular areas and neoangiogeneis. It is found in more than 90% of patients with overt SSc. Similar changes are found in patients with dermatomyositis, mixed connective tissue disease, undifferentiated connective tissue disease and they are called "scleroderma-like pattern". Absence of abnormal capillaroscopic findings can be regarded as a diagnostic criterion for primary RP. Inclusion of pathologic capillaroscopic pattern may increase the sensitivity of ACR classification criteria for SSc. In conclusion, capillaroscopy is of crucial importance for the differentiation of primary and secondary RP in rheumatic diseases, and also in differentiation between different forms of connective tissue diseases as well as for their early diagnosis. PMID- 19547980 TI - Neuropathic arthropathy progressing with multiple joint involvement in the upper extremity due to syringomyelia and type I Arnold-Chiari malformation. AB - Neuropathic arthropathy (NA), known as Charcot neuroarthropathy, is a chronic, degenerative arthropathy and is associated with decreased sensory innervation. Numerous causes of this arthropathy have been described. Neuropathic joint, although first described by Charcot in tabes dorsalis in 1868, has subsequently been observed in a variety of conditions including syringomyelia, diabetes mellitus and peripheral nerve disorders. Syringomyelia is characterized by slow progression. The shoulders and elbows are the most frequently involved joints in syringomyelia. Involvement of the hand is a quite rarely seen in the cases of NA caused by syringomyelia. In this article, we reported a case of NA secondary to syringomyelia. The characteristics of this presented case is the presence of Arnold-Chiari malformation accompanying with syringomyelia and involvement of the shoulder, elbow and hand (multiple joint involvement). PMID- 19547981 TI - Rheumatoid arthritis: scientific development from a critical point of view. AB - Rheumatoid arthritis (RA) is classified as a chronic, progressive, systemic autoimmune disorder leading to inflammation, stiffness, defective position and destruction of joints. Finally a complete loss of mobility and functioning can be the result. The fraction of disability varies strongly, for example, a systematic review shows a 50% disability in a period from first occurrence to disability from 4.5 to 22 years. Scientific efforts focused strongly on therapeutic and diagnostic methods during recent years. So far, there is no scientometric approach of the topic rheumatoid arthritis available although there is an increased need to evaluate quality and quantity of scientific research. Density equalizing algorithms, scientometric methods and large scale data analysis were applied to evaluate the quality and quantity of scientific efforts in the field of rheumatoid arthritis. Data were gained from Pubmed and ISI-Web. During the period 1901-2007, 78,128 items were published by 129 countries including the USA, UK and Germany being the most productive suppliers, representing 45.7% of all publications. Another 23 countries published more than 100 items. In terms of international cooperation the USA proved to be the most successful partner. "Arthritis and Rheumatism", "Annals of the Rheumatic Diseases" and the "Journal of Rheumatology" are the most prolific journals. The current study is the first analysis of "rheumatoid arthritis" research activities and output. Our analysis revealed single areas of interest, the most prolific journals, authors and institutions dealing with the topic. Nevertheless, statements concerning the scientific quality should be considered critical due to a bias according to self citation and co-authorship. PMID- 19547982 TI - Pathologic changes of Achilles tendon in leptin-deficient mice. AB - The objective of this study is to explore whether diabetes play roles on histopathological change of Achilles tendon in leptin-deficient mice. Ob mice (specific-pathogen free SPF) were identified at 10 days after birth and killed via dislocation of cervical spine at 12 weeks. Achilles tendon was isolated as quickly as possible and histopathological changes were investigated. Degeneration of tendinocytes, vascular proliferation, chondrocyte-like tendon cell and ruptures at insertion areas were observed. We conclude that diabetes is associated with histopathologic change in Achilles tendon of ob mice. PMID- 19547983 TI - Differentiating high-grade from low-grade chondrosarcoma with MR imaging. AB - The purpose of the study was to evaluate theMR imaging features that differentiate between low-grade chondrosarcoma (LGCS) and high-grade chondrosarcoma (HGCS) and to determine the most reliable predictors for differentiation. MR images of 42 pathologically proven chondrosarcomas (28 LGCS and 14 HGCS) were retrospectively reviewed. There were 13 male and 29 female patients with an age range of 23-72 years (average age 51 years). On MR images, signal intensity, specific morphological characteristics including entrapped fat, internal lobular architecture, and outer lobular margin, soft tissue mass formation and contrast enhancement pattern were analysed. MR imaging features used to identify LGCS and HGCS were compared using univariate analysis and multivariate stepwise logistic regression analysis. On T1-weighted images, a central area of high signal intensity, which was not seen in LGCS, was frequently observed in HGCS (n = 5, 36%) (p<0.01). Entrapped fat within the tumour was commonly seen in LGCS (n = 26, 93%), but not in HGCS (n = 1, 4%) (p<0.01). LGCS more commonly (n = 24, 86%) preserved the characteristic internal lobular structures within the tumour than HGCSs (n = 4, 29%) (p<0.01). Soft tissue formation was more frequently observed in HGCS (n = 11, 79%) than in LGCS (n = 1, 4%) (p<0.01). On gadolinium-enhanced images, large central nonenhancing areas were exhibited in only two (7.1%) of LGCS, while HGCS frequently (n = 9, 64%) had a central nonenhancing portion (p<0.01). Results of multivariate stepwise logistic regression analysis showed that soft tissue formation and entrapped fat within the tumour were the variables that could be used to independently differentiate LGCS from HGCS. There were several MR imaging features of chondrosarcoma that could be helpful in distinguishing HGCS from LGCS.Among them, soft tissue mass formation favoured the diagnosis of HGCS, and entrapped fat within the tumour was highly indicative of LGCS. PMID- 19547985 TI - Letter to the Editor re: Correctness of multi-detector-row computed tomography for diagnosing mechanical prosthetic heart valve disorders using operative findings as a gold standard. PMID- 19547986 TI - Reproducibility and changes in the apparent diffusion coefficients of solid tumours treated with combretastatin A4 phosphate and bevacizumab in a two-centre phase I clinical trial. AB - The purpose was to determine the reproducibility of apparent diffusion coefficient (ADC) measurements in a two-centre phase I clinical trial; and to track ADC changes in response to the sequential administration of the vascular disrupting agent, combretastatin A4 phosphate (CA4P), and the anti-angiogenic drug, bevacizumab. Sixteen patients with solid tumours received CA4P and bevacizumab treatment. Echo-planar diffusion-weighted MRI was performed using six b values (b = 0-750 s/mm(2)) before (x2), and at 3 and 72 h after a first dose of CA4P. Bevacizumab was given 4 h after a second dose of CA4P, and imaging performed 3 h post CA4P and 72 h after bevacizumab treatment. The coefficient of repeatability (r) of ADC total (all b values), ADC high (b = 100-750) and ADC low (b = 0-100) was calculated by Bland-Altman analysis. The ADC total and ADC high showed good measurement reproducibility (r% = 13.3, 14.1). There was poor reproducibility of the perfusion-sensitive ADC low (r% = 62.5). Significant increases in the median ADC total and ADC high occurred at 3 h after the second dose of CA4P (p < 0.05). ADC measurements were highly reproducible in a two centre clinical trial setting and appear promising for evaluating the effects of drugs that target tumour vasculature. PMID- 19547987 TI - Clinical Doppler ultrasound for the assessment of plaque ulceration in the stenosed carotid bifurcation by detection of distal turbulence intensity: a matched model study. AB - The assessment of flow disturbances due to carotid plaque ulceration may provide added diagnostic information to Doppler ultrasound (DUS) of the carotid stenosis, and indicate whether the associated hemodynamics are a potential thromboembolic source. We evaluated the effect of ulceration in a moderately stenosed carotid bifurcation on distal turbulence intensity (TI) measured using clinical DUS in matched anthropomorphic models. Several physiologically relevant ulcer geometries (hemispherical, mushroom-shaped, and ellipsoidal pointing distally and proximally) and sizes (2-mm, 3-mm and 4-mm diameter hemispheres) were investigated. An offline analysis was performed to determine several velocity based parameters from ensemble-averaged spectral data, including TI. Significant elevations in TI were observed in the post-stenotic flow field of the stenosed carotid bifurcation by the inclusion of ulceration (P < 0.001) in a region two common carotid artery diameters distal to the site of ulceration during the systolic peak and the diastolic phase of the cardiac cycle. Both the size and shape of ulceration had a significant effect on TI in the distal region (P < 0.001). Due to the use of a clinical system, this method provides the means to evaluate for plaque ulcerations in patients with carotid atherosclerosis using DUS. PMID- 19547988 TI - The role of basic fibroblast growth factor to enhance fetal intestinal mucosal cell regeneration in vivo. AB - BACKGROUND/PURPOSE: The limited application of small bowel transplantation for short bowel syndrome, mainly on the account of the morbidity and long-term implications of the procedure, has led to a search for alternative therapies. The purpose of this study was to evaluate whether basic fibroblast growth factor (bFGF) could facilitate regeneration of fetal small intestinal mucosa in vivo. METHODS: Intestinal epithelial organoid units harvested from fetal Lewis rats were injected into adult male Lewis rats whose colon was denuded of mucosa, as syngeneic recipients. One experimental group transplanted with the addition of 50 ng/ml bFGF, was compared with a control group that were transplanted without bFGF. The grafts were harvested and analyzed using histology and immunohistochemistry 3 weeks after operation. RESULTS: There were 4 anesthetic deaths, two in each group, and 11 deaths due to adhesive ileus. In no rat did neomucosa fully cover the denuded colonic muscle throughout the whole length of lumen. Histologically, the structure of the neomucosa, when present, was normal small intestinal mucosa. The small intestinal mucosa was partially restored in 100% (6 of 6) of bFGF, and in 28.6% (2 of 7) of those not given bFGF (P = 0.0021). CONCLUSIONS: These data demonstrate that bFGF can facilitate the restoration of intestinal epithelial cells, at least to some degree. Potentially, refinements of this technique could be used to facilitate the physiologic tissue engineering of small intestine in a way that allows it to move peristaltically, and have an application in the management of patients with short bowel syndrome. PMID- 19547989 TI - Anal pressure in experimental diabetes. AB - PURPOSE: Diabetes mellitus is a metabolic endocrine disorder that affects many systems, the gastrointestinal system often being among the affected systems. This experimental study work was designed to demonstrate altered anal sphincter pressures in an experimental model of diabetes mellitus (DM). MATERIALS AND METHODS: Male Wistar rats (mean weight = 250 g) were used and randomized in two groups (n = 10): CO = control and DM. DM was induced by administering a single dose of streptozotocin. Glycemic levels were measured at the start (time = 0) and end (time = 60) of the experiment and anorectal manometry at the end. RESULTS: DM rats presented a significant increase in glycemia at day 60 (DM = 407.14 +/- 73.76) as compared to the control group (time 0 = 175.7 +/- 18.62 and time 60 = 198.04 +/- 28.66). Anorectal manometry showed a significant decrease in anal pressure in the DM group at day 60 (DM = 34.2 +/- 4.97) as compared to the CO group at the same time (CO = 67.4 +/- 2.06), with P < 0.01 and P < 0.001 (Student's t test). CONCLUSIONS: The results suggest that DM, due to the high glycemic levels, lead to alterations such as anal sphincter hypotony, which may cause complications such as fecal incontinence. PMID- 19547990 TI - New approach to large haemorrhoidal prolapse: double stapled haemorrhoidopexy. AB - PURPOSE: To verify if in large haemorrhoidal prolapse (independently from the degree) in patients with no symptoms of obstructed defaecation syndrome, the use of a stapled hemorrhoidopexy variant, comprising a double stapler haemorrhoidopexy (DSH), makes it possible to reduce the percentage of failures or relapses and to standardise an objective intraoperative parameter for the purpose of quantifying internal prolapses which can then be used as a guide in determining the type of treatment to be provided. METHODS: Between June 2003 and June 2004, 353 patients were treated for haemorrhoidal prolapse. The patients suffering from large haemorrhoidal prolapse occupying more than half of the length of the anal dilator were intraoperatively selected for DSH. RESULTS: Eighty-three patients (23.5%) underwent a DSH. The degrees of the large haemorrhoidal prolapse intraoperatively selected for DSH were sub-divided as follows: 7.2% (second), 24% (third) and 68.6% (fourth). The follow-up period was 48 months. There were three cases (3.6%) of residual illnesses and five cases (6%) of a relapse. The following complications were recorded: urgency at <3 months (7.2%), haemostasis revisions (2.4%) and spontaneously draining anterior haematoma (1.2%). The results of the 270 haemorrhoidal prolapse (38 second degree, 159 third degree and 130 fourth degree) treated with the procedure for prolapse and haemorrhoids were: nine (3.3%) residual illness and 12 (4.44%) relapse illness. The following complications were recorded: urgency at <3 months (6.6%), haemostasis revisions (2.5%) and spontaneously draining anterior haematoma (0.7%). CONCLUSIONS: The intraoperative selection criterion was both efficacious and reproducible. This variant technique, which can be used in large haemorrhoidal prolapses, could allow us to further improve the quality of treatment for haemorrhoidal conditions using stapled haemorrhoidopexy, without increasing the complications. PMID- 19547992 TI - Use of hydroxyapatite ceramics for treatment of nonunited osseous defect after open fracture of lower limbs. AB - INTRODUCTION: Osseous defects in nonunited open fracture of the lower limbs are difficult to treat. Autogenous bone grafting is a promising treatment options, but a finite amount of autogenous bone graft is available from each individual and donor site morbidity remains a problem. These limitations have prompted the development and use of synthetic biomaterials such as hydroxyapatite (HA) ceramics. However, little information is available regarding the challenging cases such as nonunited open fractures The purpose of the present study is to evaluate the outcomes associated with the treatment of osseous defects in nonunited open fracture using novel HA ceramics. METHODS: Twelve bones (4 femora and 8 tibias) in 11 patients (10 men and 1 woman), with an average age of 49.1 (range 19-71) years, with nonunited osseous defects after open fracture were treated by the grafting of HA ceramics alone or with autogenous iliac bone followed by internal or external fixation. RESULTS: The patients were followed for an average of 25.2 months in average (range 10.3 -58.1 months). Finally, 11 of 12 fractures united clinically and radiographically, while one fracture required a second procedure for additional autogenous iliac bone grafting. The average time to union from the index surgery was 5.6 months (median 5.3 months, range 2.3-11 months.). Radiographs showed good incorporation of grafted HA into the host bone in most of the united cases. As a complication, transient wound drainage was found in two cases. CONCLUSION: The current protocol using the HA ceramics appears to be safe and efficacious for the treatment of osseous defects after the open fractures. PMID- 19547991 TI - Intraneuronal pyroglutamate-Abeta 3-42 triggers neurodegeneration and lethal neurological deficits in a transgenic mouse model. AB - It is well established that only a fraction of Abeta peptides in the brain of Alzheimer's disease (AD) patients start with N-terminal aspartate (Abeta(1D)) which is generated by proteolytic processing of amyloid precursor protein (APP) by BACE. N-terminally truncated and pyroglutamate modified Abeta starting at position 3 and ending with amino acid 42 [Abeta(3(pE)-42)] have been previously shown to represent a major species in the brain of AD patients. When compared with Abeta(1-42), this peptide has stronger aggregation propensity and increased toxicity in vitro. Although it is unknown which peptidases remove the first two N terminal amino acids, the cyclization of Abeta at N-terminal glutamate can be catalyzed in vitro. Here, we show that Abeta(3(pE)-42) induces neurodegeneration and concomitant neurological deficits in a novel mouse model (TBA2 transgenic mice). Although TBA2 transgenic mice exhibit a strong neuronal expression of Abeta(3-42) predominantly in hippocampus and cerebellum, few plaques were found in the cortex, cerebellum, brain stem and thalamus. The levels of converted Abeta(3(pE)-42) in TBA2 mice were comparable to the APP/PS1KI mouse model with robust neuron loss and associated behavioral deficits. Eight weeks after birth TBA2 mice developed massive neurological impairments together with abundant loss of Purkinje cells. Although the TBA2 model lacks important AD-typical neuropathological features like tangles and hippocampal degeneration, it clearly demonstrates that intraneuronal Abeta(3(pE)-42) is neurotoxic in vivo. PMID- 19547993 TI - Time-kinetic study of repigmentation in vitiligo patients by tacrolimus or pimecrolimus. AB - New topical immunomodulators have been reported to cause repigmentation of vitiligo lesions. However, time-kinetics of such repigmentation in different anatomic locations is not well known. We performed a randomized double-blind placebo control study with tacrolimus versus the vehicle and a nonrandomized control study with pimecrolimus to evaluate the time to reach significant pigmentation, its duration and extent in treated areas. Antioxidant status of serum was also assessed. Twenty patients, in the tacrolimus study, had one pair of lesions on different localizations, and 20 on face and/or upper limbs for pimecrolimus. The extent of repigmentation was evaluated by slides and mapmakings at baseline and every 4 weeks during 7 months. Adverse events were recorded. The derivatives of oxygen metabolites, the ferric reducing ability of serum and vitamin E were assessed. Three groups of patients were identified with the tacrolimus study. Eight had no significant change in response characterized by a parallel increase of repigmentation or none in treated and control areas. Nine had a better repigmentation to tacrolimus at fifth month of treatment. Three had a marked repigmentation in control areas at the end of treatment. Repigmentation was significant on the face compared to upper-limbs with pimecrolimus from fourth to seventh month. A significant reduction of oxidative stress and an increase in antioxidant capacity in serum of patients treated with topical tacrolimus was observed, while those treated with pimecrolimus did not show any significant changes but an increase in vitamin E. Our work defines three periods in repigmentation, triggering during the first 4 months, increase in pigmentation with tacrolimus and a plateau or a sustained repigmentation. The continuity of the treatment seems necessary to ensure a prolonged repigmenting effect and even an enhanced one, such as the one we observed on the face with pimecrolimus. The extent of repigmentation was more significant on the face compared to other locations probably due to differences in melanocyte density. Furthermore, we did not find any relationship between repigmentation and the duration of vitiligo. Tacrolimus was able to reduce the systemic oxidative stress independently from its repigmenting capacity. Both drugs were well tolerated. PMID- 19547994 TI - Autoantibodies to sweat glands detected by different methods in serum and in tissue from patients affected by a new variant of endemic pemphigus foliaceus. AB - Examining the patients with a new variant of endemic pemphigus foliaceus (EPF) in El Bagre, Colombia, (El Bagre-EPF), we noted several polymorphic clinical lesions around their axillary areas. Based on our clinical findings and on previous histopathological studies on the skin of these patients that showed abnormalities in their sweat glands, and the presence of mercuric selenides and iodines by autometallography assay, we decided to investigate immunoreactivity to the sweat glands in these patients. We tested for autoreactivity utilizing direct and indirect immunofluorescence (DIF, IIF). To be able to distinguish between non specific immune deposits and real autoimmune response, and knowing that sweat glands have some intrinsic autofluorescence for the presence of lipofuscin granules (that naturally fluoresce under the UV light microscope), as well as by the presence of secretory IgA, we used simultaneously immunohistochemistry (IHC). We tested ten El Bagre-EPF patients, ten healthy controls from the endemic area and ten healthy controls from the United States. We were able to visualize a specific autoreactivity to sweat glands in 8/10 cases of El Bagre-EPF by DIF, IIF and by IHC. In addition when using anti-human monoclonal antibodies to CD3, CD68, and CD20, we confirmed the presence of several specific immune responses in situ, an around the sweat glands. No healthy control cases yielded positive findings. In some chronic cases, decrease and sometimes a complete absence of sweat glands and other skin appendices was found. In addition to this, sclerodermoid changes or early sclerodermatous changes sometimes extending into the adipose tissue as a membranous lipodystrophy were observed. Autoreactivity to the neurovascular components around the sweat glands were also observed. Our data demonstrate for the first time that there is immunoreactivity toward sweat glands in El Bagre-EPF patients that seems to destroy some of these structures. PMID- 19547995 TI - Assessment of serous macular detachment in eyes with diabetic macular edema by use of spectral-domain optical coherence tomography. AB - BACKGROUND: The introduction of optical coherence tomography (OCT) has brought new potentialities for an objective evaluation of macular diseases. The purpose of the present study was to assess the serous macular detachment (SMD) in eyes with diabetic macular edema (DME) by use of spectral-domain OCT. METHODS: In this prospective study were included 79 eyes of 46 patients with diabetic retinopathy and DME. All patients underwent examination of best-corrected visual acuity (BCVA), non-contact slit-lamp fundus biomicroscopy, fluorescein angiography and OCT. Spectral-domain OCT (OCT/SLO Combination Imaging System, OPKO/OT Inc., Toronto, Ontario, Canada) was used to evaluate retinal morphology and the presence of macular traction (vitreomacular and/or from epiretinal membranes) on B-scans, C-scans and C-scan OCT/SLO fundus image overlays. With OCT were measured retinal thickness, volume, diameter of intraretinal cystoid spaces, diameter and height of SMD. The correlation of retinal thickness and volume with BCVA in all eyes with DME and the relation of SMD to retinal thickness, volume, BCVA, macular traction and ischemia were assessed. RESULTS: The SMD was diagnosed only by means of OCT in nine eyes (11.4%) of five patients. In eight of nine eyes it was combined with intermediate (300-600 microm) or severe (>600 microm) cystoid spaces, and in one eye with simple macular edema. Retinal thickness and volume correlated with BCVA (r = 0.464, P < 0.0001 and r = 0.480, P < 0.0001). The SMD height did not correlate with retinal thickness, volume or BCVA. Six eyes with SMD had macular ischemia, and five eyes had severe ischemia in retinal periphery. Macular traction was: absent in three eyes, questionable (without distortion of retinal contour from partial posterior vitreous detachment and/or epiretinal membrane) in three eyes, and definite (with distortion of retinal contour) in three eyes. CONCLUSIONS: Spectral-domain OCT provided valuable information on retinal morphology and was particularly useful in diagnosing sub-clinical SMD in eyes with DME. It disclosed the presence and strength of macular traction either by partially detached posterior hyaloid or by epiretinal membranes. C-scans and C scan OCT/SLO fundus image overlays added complementary information for the extent and location of the pathological features. Larger studies which follow subjects longitudinally are needed to explain the pathogenesis and determine the prognosis of SMD. PMID- 19547996 TI - Surgery for patients with primary hyperparathyroidism and negative sestamibi scintigraphy--a feasibility study. AB - BACKGROUND: We report the surgical treatment of a consecutive series of scan negative patients with the intention of unilateral parathyroid exploration with the aid of intraoperative quick PTH (qPTH). MATERIALS AND METHODS: The study included 35 consecutive sestamibi scan negative patients (27 women, eight men) with sporadic pHPT subjected to first time surgery. Median age was 70 years and median preoperative calcium level 2.8 mmol/L. RESULTS: Thirty-three patients had a histological diagnosis of a parathyroid adenoma (median weight 0.48 g [range 0.12 g-2.5 g]). Nineteen patients were explored bilaterally and 16 patients (46%) were operated unilaterally. The median operation time was 40 min in the unilateral group and 95 min in the bilateral group (p < 0.001). Three patients were treated for postoperative hypocalcemia after bilateral exploration versus none in the unilateral group (p = 0.23). With a minimum of 12 months of follow up, 33 patients (94.3%) were cured. One case of recurrent HPT presented after bilateral exploration with visualization of four glands. One case of persistent HPT was observed after unilateral exploration. qPTH was predictive of operative failure in both patients. CONCLUSION: Forty-six percent of the patients in our study could be operated unilaterally with a total cure rate of 94%. Patients in the unilateral group had a significant shorter operation time and a lower incidence of postoperative hypocalcemia. In conclusion our investigation shows that limited parathyroid exploration can safely be performed on patients with negative sestamibi scintigraphy by the aid of qPTH. PMID- 19547997 TI - Benefits of surgeon-performed ultrasound for primary hyperparathyroidism. AB - BACKGROUND: Focused, minimally invasive parathyroidectomy (MIP) is widely accepted when preoperative imaging localizes a single parathyroid adenoma. Many surgeons use (99m)Tc-sestamibi scintigraphy (MIBI) +/-, a cervical ultrasound for preoperative localization. We propose that surgeon-performed ultrasound (SUS) is the only imaging modality required in most patients with primary hyperparathyroidism (pHPT), resulting in patient convenience and reduced cost. MATERIALS AND METHODS: Since July 2006, patients with pHPT underwent MIP based solely on a positive SUS. Intraoperative parathyroid hormone assay was used to determine the extent of operation. A retrospective review from July 2006 through December 2008 identified 160 patients who underwent parathyroidectomy after SUS on their initial office visit. RESULTS: SUS correctly identified an enlarged parathyroid gland in 119/160 (74%) patients. In 41 patients, SUS was the only localizing study. MIBI was done in 119 patients. In 54 patients, SUS confirmed the MIBI, and in 28 patients with a negative MIBI, SUS was positive. In the 41 patients with a negative SUS, an MIBI was positive in ten. Ninety-eight patients had MIP. Theoretically, 85 MIBIs were unnecessary because of a positive SUS corresponding to a potential cost savings of at least $90,000. CONCLUSION: SUS to localize parathyroid adenomas is accurate and facilitates MIP. It provides substantial cost savings and patient convenience and should be the first diagnostic procedure performed for patients suspected to have pHPT. MIBI can be reserved for those patients in whom ultrasound has failed to localize a parathyroid gland. PMID- 19547998 TI - Differential expression of microRNA 181b and microRNA 21 in hyperplastic polyps and sessile serrated adenomas of the colon. AB - This study was designed to analyse the potential diagnostic value of miR-181b and miR-21 for discriminating hyperplastic polyps (HP) from sessile serrated adenomas (SSA) without cytologic dysplasia. Using real-time polymerase chain reaction expression levels of miR-181b and miR-21 in 18 HPs, 19 SSAs without cytologic dysplasia and 20 normal colonic mucosal specimens were examined. In addition, 20 colorectal cancers specimen were analysed for miR-181b expression. Data were normalised to RNU48 as an internal control. A differential expression of miR-181b and miR-21 was found in HPs, SSAs, and normal colonic mucosa with highest expression levels in SSAs. Levels of miR-181b but not miR-21 differed in HPs and normal mucosa. SSAs exhibited both significantly higher miR-181b levels (up to 2.01-fold; P < 0.001) and miR-21 levels (up to 1.82-fold; P = 0.011) than HPs. In contrast to HPs, SSAs are characterised by high levels of miR-181b and miR-21 expression. However, due to the overlap of values, miR-181b and miR-21 evaluation did not allow discrimination of the two lesions in every case. PMID- 19548000 TI - The number and function of circulating endothelial progenitor cells in patients with Kawasaki disease. AB - Kawasaki disease (KD) is associated with coronary artery injury. Studies have shown that the endothelial progenitor cell (EPC) participates in the process of arterial repair. Data have been reported that the number of EPC increased significantly in the subacute phase of KD. However, until now, there are no data about the functions of EPC in KD patients. The present study was designed to further investigate the number and functions of EPC in KD. Ten KD patients in the acute phase and ten healthy volunteers were recruited and attributed to the KD group and control group, respectively. The circulating CD34/kinase insert domain containing receptor double positive cells were evaluated in the two groups using flow cytometry. In vitro assays were used to measure the functions of EPC, including proliferation, adhesion, and migration activities. The plasma levels of nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha), and high sensitivity C-reactive protein (hs-CRP) were also assessed in both groups. The number of EPC in the KD group was significantly higher than that of the control group (0.021 +/ 0.007% vs. 0.014 +/- 0.003%, P < 0.05). The migratory response of EPC was significantly decreased in the KD group, compared with that of the control group (5.50 +/- 1.78 vs. 3.40 +/- 1.35 cells/high power field, P < 0.01). Similarly, the proliferative and adhesive activities of EPC in the KD group were also decreased (0.47 +/- 0.08 vs. 0.66 +/- 0.07, P < 0.01; 6.5 +/- 2.12 vs. 11.2 +/- 2.04 cells/high power field, P < 0.01). The plasma NO, TNF-alpha, and hs-CRP levels in the KD group were higher than those of the control group (54.10 +/- 11.78 vs. 38.80 +/- 11.10 mumol/l, P < 0.01; 48.20 +/- 7.42 vs. 37.00 +/- 11.12 pg/ml, P < 0.05; 87.10 +/- 30.18 vs. 5.30 +/- 3.37 mg/l, P < 0.01). The number of circulating EPC positively correlated with the level of NO (r = 0.92, P < 0.001), and the functions of EPC negatively correlated with the levels of TNF-alpha and hs-CRP, respectively. In Kawasaki disease, the number of EPC was enhanced and the functions of EPC were attenuated. The two-way regulation of circulating EPC in KD patients may be associated with the disorders of cytokines or messengers in KD patients. PMID- 19548001 TI - Increased type 3 iodothyronine deiodinase activity in a regrown hepatic hemangioma with consumptive hypothyroidism. AB - INTRODUCTION: Infantile hepatic hemangioma with consumptive hypothyroidism is a rare condition. CASE REPORT: A 4-month-old girl presented with diffuse hepatic hemangiomas during treatment of congenital hypothyroidism. Serum reverse triiodothyronine was elevated, and her hypothyroidism improved concomitant with involution of the hemangioma following prednisolone and interferon-alpha administration. At 20 months of age, 7 months after discontinuing prednisolone and interferon-alpha, a focal hemangioma regrew from one of the previous lesions and was surgically resected. The expression and activity of type 3 iodothyronine deiodinase (D3) were elevated in the resected tumor tissue compared with placenta. DISCUSSION: Here, we describe a patient with consumptive hypothyroidism and diffuse infantile hepatic hemangiomas, one of which regrew after involution following pharmacotherapy. The etiology of elevated D3 activity is also discussed. CONCLUSION: It is important to identify infantile hepatic hemangioma in patients with hypothyroidism refractory to hormone replacement therapy, who have low free triiodothyronine despite high thyrotropin and normal free thyroxine levels, and long-term follow-up will be needed for these patients. PMID- 19548002 TI - Adenovirally mediated p53 overexpression diversely influence the cell cycle of HEp-2 and CAL 27 cell lines upon cisplatin and methotrexate treatment. AB - PURPOSE: p53 gene plays a crucial role in the response to therapy. Since it is inactivated in the majority of human cancers, it is strongly believed that the p53 mutations confer resistance to therapeutics. In this paper we analyzed the influence of two mechanistically diverse antitumor agents--cisplatin and methotrexate on the proliferation and cell cycle of two head and neck squamous cancer cell lines HEp-2 (wild type p53 gene, but HPV 18/E6-inactivated protein) and CAL 27 (mutated p53 gene), along with the influence of adenovirally mediated p53 overexpression in modulation of cisplatin and methoterexate effects, whereby subtoxic vector/compound concentrations were employed. METHODS: p53 gene was introduced into tumor cells using adenoviral vector (AdCMV-p53). The cell cycle perturbations were measured by two parameter flow cytometry. The expression of p53, p21(WAF1/CIP1) and cyclin B1 proteins was examined using immunocytochemistry and western blot methods. RESULTS: In CAL 27 cells overexpression of p53 completely abrogated high S phase content observed in methotrexate-treated cells into a G1 and slight G2 arrest, while it sustained G2 arrest of the cells treated with cisplatin, along with the reduction of DNA synthesis and cyclin B1 expression. On the other hand, in HEp-2 cell line p53 overexpression slightly slowed down the progression through S phase in cells treated with methotrexate, decreased the cyclin B1 expression only after 24 h, and failed to sustain the G2 arrest after treatment with cisplatin alone. Instead, it increased the population of S phase cells that were not actively synthesizing DNA, sustained cyclin B1 expression and allowed the G2 cells to progress through mitosis. CONCLUSIONS: This study demonstrates that adenovirally mediated p53 overexpression at sub cytotoxic levels enhanced the activity of low doses of cisplatin and methotrexate in HEp-2 and CAL 27 cells through changes in the cell cycle. However, the mechanisms of these effects differ depending on the genetic context and on the chemotherapeutics' modality of action. PMID- 19548003 TI - Toxoplasma gondii: host-parasite interaction and behavior manipulation. AB - Toxoplasma gondii is an obligate intracellular parasite that causes different lesions in men and other warm-blooded animals. Humoral and cellular immune response of the host against the parasite keeps the protozoan in a latent stage, and clinical disease ensues when immunological response is compromised. Brain parasitism benefits the parasite causing behavioral changes in the host, not only in animals but also in humans. Schizophrenia and epilepsy are two neurological disorders that have recently been reported to affect humans coinfected with T. gondii. Further studies based on host-parasite interaction in several wild or domestic warm-blooded species are still necessary in order to better understand parasitism and behavioral changes caused by T. gondii. PMID- 19548004 TI - Immune response of mice with alveolar echinococcosis to therapy with transfer factor, alone and in combination with albendazole. AB - The effect of dialysable leucocyte extract (transfer factor TF) on immune response of mice infected with Echinococcus multilocularis and treated with albendazole (ABZ) was observed. TF administration increased the parasite suppressed proliferative response of T and B lymphocytes of infected mice from weeks 8 to 12 or 14 post infection (p.i.), respectively, with the most stimulative effect after TF+ABZ therapy. The CD4 T cell presence in the spleen of infected mice with TF or TF+ABZ therapy was increased from weeks 6 to 12 or 14 p.i., respectively. The production of IFN-gamma (Th1 cytokine) after TF or TF+ABZ therapy was significantly higher from weeks 6 to 12 p.i., and during this time, the significantly inhibited IL-5 synthesis (Th2 cytokine) was detected, particularly after TF+ABZ therapy. The superoxide anion (O2-) production in peritoneal macrophages of infected mice treated with TF or TF+ABZ was stimulated from weeks 8 to 18 p.i. The immunomodulative effect of TF reduced the growth of larval cysts till week 14 p.i. with a comparable intensity to the anthelmintic drug ABZ. Combined therapy TF+ABZ resulted in the greatest parasite restriction and reduced the cyst development till the end of the experiment. PMID- 19548005 TI - Metazoan parasite community of blue sea catfish, Sciades guatemalensis (Ariidae), from Tres Palos Lagoon, Guerrero, Mexico. AB - The seasonal dynamic of the metazoan parasite community of the blue sea catfish (Sciades guatemalensis) from Tres Palos Lagoon, Guerrero, Mexico, was studied at the component community and infracommunity levels. A total of 382 fish were collected during the regional dry and rainy seasons (a total of seven seasons) between April 2000 and September 2007. Nine helminths were collected: Neotetraonchus sp., Pseudoacanthostomum panamense, Austrodiplostomum compactum, Clinostomum complanatum, Metadena sp., Pseudoleptorhynchoides lamothei, Neoechinorhynchus cf. golvani, Hysterothylacium perezi, and Contracaecum sp. The infection dynamics of some dominant helminths was influenced by environmental changes generated by the dry/rainy season cycle. Nested (non-random) species composition was observed in the infracommunities during almost all of the sample period. Variation in the intensity of nestedness was attributed to a sequential colonization process over time by the dominant helminths. PMID- 19548007 TI - A comparison of analytical techniques for evaluating food waste degradation by anaerobic digestion. AB - Organic matter contained in food waste was degraded by anaerobic digestion under mesophilic and thermophilic conditions at two hydraulic retention times. Evolution of the digestion process was followed by thermogravimetry analysis, fluorescence spectroscopy and (1)H nuclear magnetic resonance. All analytical methods suggested that longer retention times might be required for food waste stabilization under mesophilic conditions as compared to thermophilic stabilization. All the analytical methods showed that the stabilization process consisted of two steps, where complex organic molecules were formed during initial stabilization and then digested providing sufficient hydraulic retention time. Longer hydraulic retention times were required for food waste stabilization under mesophilic conditions. Overall, thermal and (1)H NMR analyses of the digestate samples might be recommended if more detailed analysis is required, while fluorescence measurements can be used as a fast screening technique, which provides qualitative assessment of the stabilization process. PMID- 19548008 TI - Analysis of a continuous culture of Fibrobacter succinogenes S85 on a standardized glucose medium. AB - Continuous cultures of Fibrobacter succinogenes S85 were performed on a standardized fully synthetic culture medium with glucose as carbon source at a dilution rate (D = 0.02 h(-1)) in a 5-L bioreactor. The culture was stabilized during 20 days and demonstrated the ability of Fibrobacter succinogenes to grow in this synthetic medium. CO(2) partial pressure and redox potential probes were used to check the anaerobic state of the culture. The biomass yield was calculated 0.206 g (g glucose)(-1) and the production yield of succinate, the major end-product, was 0.63 mol (mol glucose)(-1). The consistency of the experimental data was checked by proton and mass (C, N) balances. The results were satisfactory (90-110% recovery) leading to derive a stoichiometric equation representative of the growth on glucose. The stoichiometric coefficients were calculated using data reconciliation and linear algebra methods enabling to obtain a complete modeling of all conversion yields possible. PMID- 19548009 TI - The use of additives as the stimulator on mycelial biomass and exopolysaccharide productions in submerged culture of Grifola umbellata. AB - In this study, various additives including organic acids, alcohols, vegetable oils, surfactants and polymers were added in the cultural medium to investigate their stimulatory effects on Grifola umbellate mycelia growth and exopolysaccharide (EPS) production. It was found that the commonly used stimulatory additives, effective in other mushrooms' cultures, exhibited negative results in Grifola umbellata submerged culture. In contrast, the polymer additive, polyethylene glycol (PEG), displayed an effective stimulatory effect on both biomass and EPS productions. With the addition of PEG8 (molecular weight: 8,000 Da), the mycelial biomass production at day 12 was increased from 4.69 to 6.30 g/L, accounting for a 34% increase. Meanwhile, the EPS production was enhanced from 0.478 to 0.767 g/L, accounting for 60% increase. PMID- 19548010 TI - Clinical and microbiological performance of resin-modified glass-ionomer liners after incomplete dentine caries removal. AB - The aims of this study were to evaluate clinically and microbiologically the effects of two resin-modified glass-ionomer cements (RMGICs) used as liners after incomplete dentine caries removal and to identify Streptococcus mutans and Streptococcus sobrinus strains isolated from dentine samples, before and after indirect pulp treatment. Twenty-seven primary molars with deep carious lesions, but without signs and symptoms of irreversible pulpitis, were submitted to indirect pulp treatment. Treatment consisted of incomplete excavation of the carious dentine, application of one of the RMGICs (Vitrebond or Fuji Lining LC) or calcium hydroxide cement (Dycal), and sealing for 3 months. Clinical evaluation (consistency, color, and wetness of dentine) and carious dentine collects were performed before temporary sealing and after the experimental period. Microbiological samples were cultivated in specific media for subsequent counting of mutans streptococci (MS) and lactobacilli (LB). MS colonies were selected for identification of S. mutans and S. sobrinus by polymerase chain reaction. After 3 months, the remaining dentine was hard and dry, and there was a significant decrease in the number of MS and LB, in all groups, although complete elimination was not achieved in 33% and 26% of the teeth for MS and LB, respectively. From 243 MS colonies selected, 216 (88.9%) were identified as S. mutans and only 2 (0.8%) as S. sobrinus. The use of resin-modified glass-ionomer liners after incomplete caries removal, as well as a calcium hydroxide cement, promoted significant reduction of the viable residual cariogenic bacteria in addition to favorable clinical changes in the remaining carious dentine. PMID- 19548011 TI - Plaque removal efficacy of power and manual toothbrushes: a comparative study. AB - The aim of this study was to compare the plaque removal efficacy of a new oscillating/rotating/pulsating toothbrush [Oral-B Professional Care 8500 (PC 8500)] with two manual toothbrushes [Oral-B CrossAction Vitalizer (CAV) and Oral B Indicator (IND), respectively]. The safety of the PC 8500 was also assessed. The study was a single-use, observer-masked, randomised 3 x 3 Latin square crossover design balanced for carryover effects. The enrolled subjects (n = 66) refrained from brushing for 23-25 h before each clinical examination. Plaque scores were recorded before and after brushing with the allocated toothbrush using the Turesky et al. modification of the Quigley and Hein plaque index. The safety was assessed evaluating the soft tissue conditions present after 30 days of the use of the PC 8500. The PC 8500 toothbrush was better in plaque removal efficacy compared with the CAV and IND brushes for full mouth and approximal surfaces (P < 0.01). When marginal surfaces were considered, the PC 8500 was significantly more effective than the IND (P < 0.01). No significant differences were found between PC 8500 and CAV (P > 0.05). The latter was shown to be significantly more effective than the IND at all tooth surfaces (P < 0.01). Safety examinations revealed the onset of only two small gingival abrasions after the 30-day use of the PC 8500. The PC 8500 toothbrush demonstrated to be more effective in plaque control than the CAV and IND in the full mouth and approximal surfaces and similar to the CAV in the marginal surfaces. The PC 8500 was safe to oral tissues in long-term use. PMID- 19548012 TI - Oral malodorous compound activates mitochondrial pathway inducing apoptosis in human gingival fibroblasts. AB - Hydrogen sulfide (H(2)S) is a main cause of physiologic halitosis. H(2)S induces apoptosis in human gingival cells, which may play an important role in periodontal pathology. Recently, it has been reported that H(2)S induced apoptosis and DNA damage in human gingival fibroblasts (HGFs) by increasing the levels of reactive oxygen species. However, the mechanisms of H(2)S-induced apoptosis have not been clarified in HGFs. The objective of this study was to determine the apoptotic pathway activated by H(2)S in HGFs. The HGFs were exposed to 50 ng/mL H(2)S, resulting in 18 ng/mL in the culture medium, which is lower than the concentration in periodontal pockets. The number of apoptotic cells after 24 and 48 h incubation was significantly higher than that in the control cultures (p < 0.05). Mitochondrial membrane depolarization and the release of cytochrome c, and caspase-3, and caspase-9 were also significantly increased after both 24- and 48-h incubation (p < 0.05), whereas caspase-8, a key enzyme in the receptor ligand-mediated pathway causing apoptosis, was not activated. The present study shows that H(2)S triggered the mitochondrial pathway causing apoptosis in HGFs but did not activate the receptor ligand-mediated pathway. PMID- 19548013 TI - COL25A1 triggers and promotes Alzheimer's disease-like pathology in vivo. AB - Collagen XXV alpha 1 (COL25A1) is a collagenous type II transmembrane protein purified from senile plaques of Alzheimer's disease (AD) brains. COL25A1 alleles have been associated with increased risk for AD in a Swedish population. COL25A1 is specifically expressed in neurons and binds to aggregated Abeta in vitro. However, its contribution to the pathogenesis of AD and in vivo function are unknown. Here, we report that over-expression of COL25A1 in transgenic mice increases p35/p25 and beta-site APP-cleaving enzyme 1 (BACE1) levels, facilitates intracellular aggregation and extracellular matrix deposits of Abeta, and causes synaptophysin loss and astrocyte activation. COL25A1 mice displayed reduced anxiety-like behavior in elevated plus maze and open field tests and significantly slower swimming speed in Morris water maze. In stable cell lines, motifs in noncollagenous domains of COL25A1 were important for the induction of BACE1 expression. These findings demonstrate that COL25A1 leads to AD-like pathology in vivo. Modulation of COL25A1 function may represent an alternative therapeutic intervention for AD. PMID- 19548014 TI - Neurosurgery in Italy: the past, the present, the future. PMID- 19548015 TI - Paradoxical robust visual evoked potentials in young patients with cortical blindness. AB - The objective of this study was to review retrospectively cases of clinically blind children in whom robust pattern visual evoked potentials (VEPs) were recorded. VEP records from a 10-year period (1990-2000) were reviewed. We searched for charts of children who were clinically cortically blind, but in whom assessment of visual acuity, using visual evoked potentials (VEPs), was normal or close to normal. The majority (77.5%) of VEP and behavioral acuity measures were concordant (subset analysis). Of the 1,113 VEP records, 9 cases (<1% of records reviewed) had clinically compromised vision with fair to good levels of visual function using VEPs. The commonality among the cases was the presence of suspected cortical visual impairment with seizures and developmental delay. VEP acuity cannot be correlated unequivocally with visually guided behaviour. In specific cases, particularly cases with developmental delay and neuroradiographic abnormalities, a child who is behaviorally blind with no clinical evidence of vision may show robust VEPs even to small patterns. This finding might be consistent with a defect of the visual association cortex. PMID- 19548016 TI - Estimation of the cancer risk to humans resulting from the presence of cyclophosphamide and ifosfamide in surface water. AB - BACKGROUND, AIM, AND SCOPE: Anti-tumour agents and their metabolites are largely excreted into effluent, along with other pharmaceuticals. In the past, investigations have focused on the input and analysis of pharmaceuticals in surface and ground water. The two oxazaphosphorine compounds, cyclophosphamide and ifosfamide are important cytostatic drugs used in the chemotherapy of cancer and in the treatment of autoimmune diseases. Their mechanism of action, involving metabolic activation and unspecific alkylation of nucleophilic compounds, accounts for genotoxic and carcinogenic effects described in the literature and is reason for environmental concern. The anti-tumour agents cyclophosphamide (CP) and ifosfamide (IF) were not biodegraded in biodegradation tests. They were not eliminated in municipal sewage treatment plants. Degradation by photochemically formed HO radicals may be of some relevance only in shallow, clear, and nitrate rich water bodies but could be further exploited for elimination of these compounds by advanced oxidation processes, i.e. in a treatment of hospital waste water. Therefore, CP and IF are assumed to persist in the aquatic environment and to enter drinking water via surface water. The risk to humans from input of CP and IF into surface water is not known. MATERIALS AND METHODS: The local and regional, i.e. nationwide predicted environmental concentration (PEC(local), PEC(regional)) of CP and IF was calculated for German surface water. Both compounds were measured in hospital effluents, and in the influent and effluent of a municipal treatment plant. Additionally, published concentrations in the effluent of sewage treatment plants and surface water were used for risk assessment. Excretion rates were taken into account. For a worst-case scenario, maximum possible ingestion of CP or IF by drinking 2 L a day of unprocessed surface water over a life span of 70 years was calculated for adults. Elimination in drinking water processing was neglected, as no data is available. This intake was compared with intake during anti-cancer treatment. RESULTS AND DISCUSSION: Intake of CP and IF for anti-cancer treatment is typically 10 g within a few months. Under such conditions, a relative risk of 1.5 for the carcinogenic compounds CP and IF is reported in the literature. In the worst case, the maximum possible intake by drinking water is less than 10(-3) (IF) and 10(-5) (CP) of this amount, based on highest measured local concentrations. On a nationwide average, the factor is approx. 10(-6) or less. CONCLUSIONS: The additional intake of CP and IF due to their emission into surface water and its use without further treatment as drinking water is low compared to intake within a therapy. This approach has shortcomings. It illustrates the current lack of methodology and knowledge for the specific risk assessment of carcinogenic pharmaceuticals in the aquatic environment. IF and CP are directly reacting with the DNA. Therefore, with respect to health effects a safe threshold concentration for these compounds cannot be given. The resulting risk is higher for newborns and children than for adults. Due to the lack of data the risk for newborns and children cannot be assessed fully. The data presented here show that according to present knowledge the additional risk of cancer cannot be fully excluded, especially with respect to children. Due to the shortage of data for effects of CP and IF in low doses during a whole lifespan, possible effects were assessed using data of high doses of CP and IF within short-term ingestion, i.e. therapy. This remains an unresolved issue. Anyway, the risk assessment performed here could give a rough measure of the risks on the one hand and the methodological shortcomings on the other hand which are connected to the assessment of the input of genotoxic and carcinogenic pharmaceuticals such as CP and IF into the aquatic environment. Therefore, we recommend to take measures to reduce the input of CP and IF and other carcinogenic pharmaceuticals. We hope that our manuscript further stimulates the discussion about the human risk assessment for carcinogenic pharmaceuticals in the aquatic environment. RECOMMENDATIONS AND PERSPECTIVES: CP and IF are carcinogens. With respect to newborn and children, reduction of the emission of CP and IF into effluent and surface water is recommended at least as a precautionary measure. The collection of unused and outdated drugs is a suitable measure. Collection of patients' excreta as a measure of input reduction is not recommended. Data suitable for the assessment of the risk for newborn and children should be collected in order to perform a risk assessment for these groups. This can stimulate discussion and give new insights into risk assessment for pharmaceuticals in the environment. Our study showed that in the long term, effective risk management for the reduction of the input of CP and IF are recommendable. PMID- 19548017 TI - Effects of shoreline sensitivity on oil spill trajectory modeling of the Lower Mississippi River. AB - BACKGROUND, AIM, AND SCOPE: The Lower Mississippi River is a major transportation route for commercial goods and petroleum products produced and refined locally. Oil spills caused by vessel accidents and equipment failure at refineries are a serious threat to the drinking water supply of Southern Louisiana, as well as to the many natural, economic, and social resources supported by the river. Providing accurate trajectory modeling to contingency planners is critical to protecting the local environment. The majority of trajectory model results, assuming a uniform shoreline, show 60-70% of spilled oil can be retained. This study examines the impact of detailed shoreline mapping that captures spatial and temporal changes in shoreline type on oil spill trajectory modeling. MATERIALS AND METHODS: Detailed shoreline maps based on recent remote sensing imagery were generated to identify spatial changes in shoreline. A hydrodynamic model of the 78 mile reach from Convent, Louisiana to West Pointe a la Hache was developed to obtain the stage levels and velocity fields of four river discharges. Based on river stage level, another layer was added to the shoreline maps, so that shoreline type was accurately represented at each river discharge, a feature not included in previous mapping. An oil spill trajectory model was then used to investigate the effect of implementing different re-floatation half-lives that correlate to the shoreline maps developed for this study at four river discharges. RESULTS: Detailed shoreline mapping showed the Lower Mississippi River has four major shoreline types each with different oil re-floatation half lives: muddy clay, sand, low vegetation, and high vegetation. As flow rate changed, the shoreline spatial variability also changed, from 84% mud/sand and 16% vegetation at low flow rates to 4% mud and 96% vegetation at higher flow rates. At flow rates with large variability in shoreline type, the distribution of oil attached to the shore was significantly different from results of simulations that used a constant shoreline type and re-floatation half-life. DISCUSSION: At low flow rates, simulations with the detailed delineation of shoreline type predicted that approximately 30% of the oil would be beached/retained because the oil was able to travel further down the reach and interact with the shoreline in multiple locations. Simulations at the low flow rates with the existing shoreline mapping predicted approximately 65% of the oil would be retained as did all the simulations at the highest flow rates. At high flow rates, the oil interacted mostly with vegetation and results were very similar to those obtained with a single re-floatation half-life of 1 year. In addition to shoreline type, river geometry and the hydrodynamics were major factors influencing the distribution of oil along the river reach. CONCLUSIONS: Shoreline re-floatation half-lives have a major impact on simulating the distribution of oil along the shore after a spill, especially in areas with a high variability of shoreline type as in the lower Mississippi River. Assigning the correct re-floatation half-life and retention capacity is only possible when shoreline types have been correctly identified. The maps developed for this study provided an important level of detail and incorporated the change in shoreline type with flow rate, resulting in more detailed trajectory modeling of the study reach. RECOMMENDATIONS AND PERSPECTIVES: Shoreline maps should include as much detail about shoreline type as possible. When developing shoreline maps or environmental sensitivity assessments, the focus should include specific characteristics of the study area; using standardized maps or methods of assessment may leave out detail that could negatively impact modeling efforts. Finally, shoreline sensitivity to oiling is an important area of research that will benefit from an improved understanding of oil retention by vegetation. PMID- 19548018 TI - Overexpression of a FAD3 desaturase increases synthesis of a polymethylene interrupted dienoic fatty acid in seeds of Arabidopsis thaliana L. AB - A cDNA encoding the Arabidopsis extraplastidic linoleate desaturase (FAD3) was overexpressed in the seeds of wild-type Arabidopsis and in a mutant line that accumulates high levels of oleic acid. In the transformed wild-type plants, linolenic acid (18:3Delta9,12,15) increased from 19% to nearly 40% of total seed fatty acids, with a corresponding decrease in linoleate content (18:2Delta9,12). In the high oleate mutant, a large increase in the level of a fatty acid identified by gas-chromatography/mass-spectrometry as mangiferic acid (18:2Delta9,15) was observed. The results demonstrate that the polymethylene interrupted dienoic fatty acid, mangiferic acid, can be produced in seed oil through the overexpression of a fatty acid n-3 desaturase. PMID- 19548019 TI - Bone turnover in bone biopsies of patients with low-energy cortical fractures receiving bisphosphonates: a case series. AB - Recent reports of long-term bisphosphonate-treated patients developing cortical fractures have raised concerns that such fractures may relate to excessive suppression of bone turnover after prolonged use of these drugs. To evaluate the bone histology of patients presenting with cortical fractures after bisphosphonate therapy, we conducted a retrospective analysis of patients treated at Washington University Bone Health Program presenting with a history of low energy cortical fractures (femoral shaft, pelvis, rib, metatarsal, and ankle), who had received bisphosphonates for at least two consecutive years and had undergone bone biopsy. Fifteen of 54 patients who underwent bone biopsy between November 2004 and March 2007 met the criteria. Of these, 10 patients had findings of suppressed trabecular bone remodeling, as demonstrated by lack of double tetracycline labels. There were no significant differences in bone density, clinical features, and biochemical features between those with suppressed turnover and the other five subjects with normal remodeling. However, the low turnover group had received bisphosphonates (primarily alendronate) for a significantly longer duration (6.5 +/- 0.6 vs. 3.9 +/- 0.8 years, P = 0.02). Thus, about two-thirds of patients presenting with cortical fractures while on long-term treatment with bisphosphonates had suppressed turnover. Since the prevalence of such histological findings in nonfracture patients remains unknown, the impact of suppressed bone turnover on the development of cortical fractures cannot be determined. Considering the widespread use of bisphosphonates, it appears that the overall risk of cortical fractures is low. However, there may be a subset of as yet unidentified patients who could be predisposed to this complication. PMID- 19548021 TI - Environmental integrated assessment via Monte Carlo simulation with a case study of the Mid-Atlantic region, USA. AB - Environmental integrated assessments are often carried out via the aggregation of a set of environmental indicators. Aggregated indices derived from the same data set can differ substantially depending upon how the indicators are weighted and aggregated, which is often a subjective matter. This article presents a method of generating aggregated environmental indices in an objective manner via Monte Carlo simulation. Rankings derived from the aggregated indices within and between three Monte Carlo simulations were used to evaluate the overall environmental condition of the study area. Other insights, such as the distribution of good or bad values of indicators at a watershed and/or a subregion, were observed in the study. PMID- 19548020 TI - Bioinformatic characterization of p-type ATPases encoded within the fully sequenced genomes of 26 eukaryotes. AB - P-type ATPases play essential roles in numerous processes, which in humans include nerve impulse propagation, relaxation of muscle fibers, secretion and absorption in the kidney, acidification of the stomach and nutrient absorption in the intestine. Published evidence suggests that uncharacterized families of P type ATPases with novel specificities exist. In this study, the fully sequenced genomes of 26 eukaryotes, including animals, plants, fungi and unicellular eukaryotes, were analyzed for P-type ATPases. We report the organismal distributions, phylogenetic relationships, probable topologies and conserved motifs of nine functionally characterized families and 13 uncharacterized families of these enzyme transporters. We have classified these proteins according to the conventions of the functional and phylogenetic IUBMB-approved transporter classification system ( www.tcdb.org , Saier et al. in Nucleic Acids Res 34:181-186, 2006; Nucleic Acids Res 37:274-278, 2009). PMID- 19548022 TI - Willingness to pay for improving the residential waste disposal system in Korea: a choice experiment study. AB - This study attempts to apply choice experiments with regard to the residential waste disposal system (RWDS) in Korea by considering various attributes that are related to RWDS. Using data from a survey conducted on 492 households, the empirical analysis yields estimates of the willingness to pay for a clean food waste collection facility, the collection of small items (such as obsolete mobile phones and add-ons for personal computers), and a more convenient large waste disposal system. The estimation results of multinomial logit models are quite similar to those of nested logit models. The results reveal that residents have preferences for the cleanliness of facilities and the collection of small items. In Korea, residents are required to purchase and attach stickers for the disposal of large items; they want to be able to obtain stickers at not only village offices but also supermarkets. On the other hand, the frequency of waste collection is not a significant factor in the choice of the improved waste management program. PMID- 19548023 TI - Climate change adaptation for the US National Wildlife Refuge System. AB - Since its establishment in 1903, the National Wildlife Refuge System (NWRS) has grown to 635 units and 37 Wetland Management Districts in the United States and its territories. These units provide the seasonal habitats necessary for migratory waterfowl and other species to complete their annual life cycles. Habitat conversion and fragmentation, invasive species, pollution, and competition for water have stressed refuges for decades, but the interaction of climate change with these stressors presents the most recent, pervasive, and complex conservation challenge to the NWRS. Geographic isolation and small unit size compound the challenges of climate change, but a combined emphasis on species that refuges were established to conserve and on maintaining biological integrity, diversity, and environmental health provides the NWRS with substantial latitude to respond. Individual symptoms of climate change can be addressed at the refuge level, but the strategic response requires system-wide planning. A dynamic vision of the NWRS in a changing climate, an explicit national strategic plan to implement that vision, and an assessment of representation, redundancy, size, and total number of units in relation to conservation targets are the first steps toward adaptation. This adaptation must begin immediately and be built on more closely integrated research and management. Rigorous projections of possible futures are required to facilitate adaptation to change. Furthermore, the effective conservation footprint of the NWRS must be increased through land acquisition, creative partnerships, and educational programs in order for the NWRS to meet its legal mandate to maintain the biological integrity, diversity, and environmental health of the system and the species and ecosystems that it supports. PMID- 19548024 TI - Erosional consequence of saltcedar control. AB - Removal of nonnative riparian trees is accelerating to conserve water and improve habitat for native species. Widespread control of dominant species, however, can lead to unintended erosion. Helicopter herbicide application in 2003 along a 12 km reach of the Rio Puerco, New Mexico, eliminated the target invasive species saltcedar (Tamarix spp.), which dominated the floodplain, as well as the native species sandbar willow (Salix exigua Nuttall), which occurred as a fringe along the channel. Herbicide application initiated a natural experiment testing the importance of riparian vegetation for bank stability along this data-rich river. A flood three years later eroded about 680,000 m(3) of sediment, increasing mean channel width of the sprayed reach by 84%. Erosion upstream and downstream from the sprayed reach during this flood was inconsequential. Sand eroded from channel banks was transported an average of 5 km downstream and deposited on the floodplain and channel bed. Although vegetation was killed across the floodplain in the sprayed reach, erosion was almost entirely confined to the channel banks. The absence of dense, flexible woody stems on the banks reduced drag on the flow, leading to high shear stress at the toe of the banks, fluvial erosion, bank undercutting, and mass failure. The potential for increased erosion must be included in consideration of phreatophyte control projects. PMID- 19548025 TI - Hydrogeochemical indicators of groundwater flow systems in the Yangwu River alluvial fan, Xinzhou Basin, Shanxi, China. AB - Based on analysis of groundwater hydrochemical and isotopic indicators, this article aims to identify the groundwater flow systems in the Yangwu River alluvial fan, in the Xinzhou Basin, China. Groundwater delta(2)H and delta(18)O values indicate that the origin of groundwater is mainly from precipitation, with local evaporative influence. d-excess values lower than 10% in most groundwaters suggest a cold climate during recharge in the area. Major ion chemistry, including rCa/rMg and rNa/rCl ratios, show that groundwater salinization is probably dominated by water-rock interaction (e.g., silicate mineral weathering, dissolution of calcite and dolomite and cation exchange) in the Yangwu River alluvial fan, and locally by intensive evapotranspiration in the Hutuo River valley. Cl and Sr concentrations follow an increasing trend in shallow groundwater affected by evaporation, and a decreasing trend in deep groundwater. (87)Sr/(86)Sr ratios reflect the variety of lithologies encountered during throughflow. The groundwater flow systems (GFS) of the Yangwu River alluvial fan include local and intermediate flow systems. Hydrogeochemical modeling results, simulated using PHREEQC, reveal water-rock interaction processes along different flow paths. This modeling method is more effective for characterizing flow paths in the intermediate system than in the local system. Artificial exploitation on groundwater in the alluvial fan enhances mixing between different groundwater flow systems. PMID- 19548026 TI - A change in practice from epidural to intrathecal morphine analgesia for hepato pancreato-biliary surgery. AB - BACKGROUND: This study was designed to audit the change of anesthetic practice from thoracic epidural analgesia (TEA) to intrathecal morphine (ITM) combined with patient-controlled analgesia (PCA) for hepato-pancreato-biliary (HPB) surgery. METHODS: All patients who underwent major HPB surgery and received TEA or ITM from March 2005 to March 2008 were identified. Patients who received PCA alone were used for comparison. Data were retrospectively collected and analyzed for success of TEA, perioperative intravenous fluid (IVF) volume administered, hypotension, complications, and hospital stay. RESULTS: During the study period, 51 (32%) patients received TEA, 79 (49%) received ITM plus PCA opiate, and 31 (19%) received PCA alone. The incidence of postoperative hypotension was significantly higher in those who received TEA compared with those who received ITM (21/51 (41%) vs. 7/79 (9%), P < 0.001). The median (range) perioperative IVF administration was higher in the TEA group compared with the ITM group for both the first 24 h (6 (3-11) liters vs. 5 (3-11) liters, P < 0.05) and in total (15.5 (5-48.5) liters vs. 9 (3-70) liters, P < 0.001). Respiratory complications occurred in five (10%) of the TEA group compared with one (1%) in the ITM group (P < 0.05). The median (range) hospital stay was longer in the TEA group compared with the ITM group (9 (3-36) days vs. 7 (3-55) days, P < 0.01). CONCLUSIONS: In a resource-limited setting, ITM, compared with TEA, is associated with a reduced incidence of postoperative hypotension, reduced IVF requirements, shorter hospital stay, and lowers the incidence of respiratory complication. PMID- 19548027 TI - An update on long-term outcome of curative hepatic resection for hepatocholangiocarcinoma. AB - BACKGROUND: Hepatocholangiocarcinoma (HCC-CC) is a rare primary liver cancer. Its long-term prognosis is still not well-defined. Results from the Eastern and Western literature have been conflicting and no conclusions can be drawn. The aim of the present study was to review the long-term outcome of curative hepatectomy for HCC-CC. PATIENTS AND METHODS: Prospectively collected data from December 1991 to 2006 recording patients with primary liver cancer receiving curative hepatectomy were reviewed. Twenty-five patients, 16 men and 9 women with a median age of 48 years, all ethnic Chinese, had HCC-CC. Their long-term outcome of resection was analyzed and compared to that of patients with cholangiocarcinoma (CC) or hepatocellular carcinoma (HCC). RESULTS: The HCC-CC patients had a median tumor size of 7.5 cm. Five of them developed postoperative complications. The median follow-up period was 25 months. All of the patients developed recurrence. The median overall survival was 25.2 months. The HCC-CC and CC groups had significantly worse overall survival than the HCC group (HCC versus HCC-CC, p = 0.012; HCC versus CC, p = 0.001) whereas between them there was no significant difference (p = 0.822). As for disease-free survival, there was no significant difference between the three groups; the median disease-free survival for HCC-CC patients was 13.5 months; that for CC patients, 16.1 months; and that for HCC patients, 19.0 months. All HCC-CC patients died within 120 months of primary surgery. CONCLUSIONS: Hepatocholangiocarcinoma entails poor long-term outcome after potentially curative hepatectomy. Other modalities of treatment should be explored in order to prolong survival of patients with this disease. PMID- 19548028 TI - Evaluation of double tract reconstruction after total gastrectomy in patients with gastric cancer: prospective randomized controlled trial. AB - BACKGROUND: The double tract (DT) method was compared with the Roux-en-Y (R-Y) method to identify the optimal reconstruction procedure after total gastrectomy for patients with gastric cancer. The DT reconstruction is as simple as the R-Y, and it can be safely performed even after total gastrectomy. However, these have been no studies evaluating the usefulness of DT reconstruction in comparison to R Y reconstruction. METHODS: A group of 44 patients with gastric cancer were intraoperatively randomized for R-Y (n = 23) or DT reconstruction (n = 21) after total gastrectomy (TG). Body weight, food intake, nutritional conditions, and quality of life (QOL) were determined at 3 and 12 months after the operation. This study is registered with ClinicalTrials.gov, no. NCT00746161. RESULTS: Food intake significantly decreased soon after the operation. No differences were observed between the DT and R-Y groups. The body weight decreased throughout the ensuing period (P < 0.05) and thereafter gradually recovered. However, no differences were observed between the two groups. Among the nutritional laboratory parameters, serum prealbumin, retinol-binding protein, total cholesterol, and triglyceride were decreased soon after the operation. The changes of those parameters were not substantially different between the two groups. The postoperative QOL was evaluated, and no differences were observed between those groups. CONCLUSIONS: There were no particular advantages in the DT method after TG in comparison to the simple R-Y method in terms of body weight, QOL, and nutritional conditions, suggesting that the DT method might not be recommended after TG for patients with gastric cancer. PMID- 19548029 TI - Annuloaortic ectasia and ascending aortic aneurysm as a cardiovascular manifestation of acromegaly. PMID- 19548030 TI - Experimental study on biopsy sampling using new flexible cryoprobes: influence of activation time, probe size, tissue consistency, and contact pressure of the probe on the size of the biopsy specimen. AB - Cryoextraction is a procedure for recanalization of obstructed airways caused by exophytic growing tumors. Biopsy samples obtained with this method can be used for histological diagnosis. The objective of this study was to evaluate the parameters influencing the size of cryobiopsies in an in vitro animal model. New flexible cryoprobes with different diameters were used to extract biopsies from lung tissue. These biopsies were compared with forceps biopsy (gold standard) in terms of the biopsy size. Tissue dependency of the biopsy size was analyzed by comparing biopsies taken from the lung, the liver, and gastric mucosa. The effect of contact pressure exerted by the tip of the cryoprobe on the tissue was analyzed on liver tissue separately. Biopsy size was estimated by measuring the weight and the diameter. Weight and diameter of cryobiopsies correlated positively with longer activation times and larger diameters of the cryoprobe. The weight of the biopsies was tissue dependent: lung < liver < stomach. Only little tissue dependency was found for the biopsy diameter. The biopsy size increased when the probe was pressed on the tissue during cooling. Cryobiopsies can be taken from different tissue types with flexible cryoprobes. The size of the samples depends on tissue type, probe diameter, application time, and pressure exerted by the probe on the tissue. Even the cryoprobe with the smallest diameter can provide larger biopsies than a forceps biopsy in lung. It can be expected that the same parameters influence the sample size of biopsies in vivo. PMID- 19548031 TI - Dobutamine enhances alveolar fluid clearance in a rat model of acute lung injury. AB - Acute lung injury (ALI) and its most severe form, acute respiratory distress syndrome (ARDS), remain leading factors for morbidity and mortality in critically ill patients. A significant aspect of ALI and ARDS is impaired alveolar fluid clearance (AFC). Improvements in therapies for these types of respiratory illnesses will require an understanding of the mechanisms that control AFC. The present study was designed to determine whether the administration of dobutamine decreases pulmonary edema and stimulates AFC in a rat model of lipopolysaccharide induced lung injury. Adult male Sprague-Dawley rats were randomly divided into three groups: control, lipopolysaccharide, and lipopolysaccharide + dobutamine. The effect of dobutamine on AFC and the expression of aquaporin-1 and aquaporin-5 were examined. Lipopolysaccharide administration results in significant lung injury with impaired AFC, while dobutamine improves alveolar fluid reabsorption with elevation of aquaporin-1 and aquaporin-5. Our study indicates that dobutamine may enhance alveolar fluid reabsorption by increasing the expression of aquaporin-1 and aquaporin-5. PMID- 19548032 TI - Timing means everything, maybe. PMID- 19548033 TI - Hepatic stellate cells may relate to progression of intrahepatic cholangiocarcinoma. AB - BACKGROUND: Although cumulative evidence supports the fact that stromal myofibroblasts promote tumor progression, the influence of myofibroblasts on intrahepatic cholangiocarcinoma (ICC) is unclear. We hypothesized that hepatic stellate (HS) cells can differentiate into myofibroblasts in ICC stroma and that they promote cancer progression. This study aims to: (1) assess the influence of myofibroblasts on the prognosis of ICC, (2) identify HS cells in ICC stroma, and (3) investigate the interaction between HS cells (LI90 and LX-2) and ICC cells (HuCCT-1 and MEC) in vitro. METHODS: The association between alpha-smooth muscle actin (alpha-SMA) expression and the prognoses of 46 ICC patients after hepatic resection was evaluated by immunohistochemical analysis. The HS cells in myofibroblasts of ICC were identified (double immunostaining) using antibodies for alpha-SMA, glial fibrillary acidic protein (GFAP), and desmin. The influence of HS cells on the invasion and growth of ICC cells was examined in vitro using a coculture system. RESULTS: Patients with high alpha-SMA expression exhibited the worse outcomes. Multivariate analyses revealed that high alpha-SMA expression (P = 0.0045) and positivity for lymph-node metastasis were independent prognostic factors. Because desmin- or GFAP-positive cells coexpressing alpha-SMA were observed in the ICC samples, they were considered to be derived from the HS cells. On coculturing with HS cells, a remarkable increase was observed in the invasion and growth of the two ICC cell lines. CONCLUSIONS: Stromal myofibroblasts may relate to the poor prognoses in ICC patients. HS cells appear to be involved in the progression of ICC. PMID- 19548034 TI - Involvement of nigrostriatal pathway in Japanese encephalitis with movement disorders: evidence from 99mTc-TRODAT-1 and 123I-IBZM SPECT imagings. AB - PURPOSE: The purpose of this study was to evaluate molecular evidence of nigrostriatal pathway involvement in Japanese encephalitis (JE) survivors with movement complications. METHODS: Three JE patients were recruited. All had cranial magnetic resonance imaging (MRI) and single-photon emission computed tomography (SPECT) studies with (99m)Tc-TRODAT-1 and (123)I-IBZM. RESULTS: Cranial MRI revealed involvement of bilateral thalami, substantia nigra, and medial temporal lobes in all three patients, but only case 1 had additional bilateral basal ganglia involvement. The (99m)Tc-TRODAT-1 SPECT for presynaptic dopamine transporter imaging disclosed asymmetrical decreases in bilateral striatal uptake in all three patients. However, the (123)I-IBZM SPECT imaging for postsynaptic D2 dopamine receptors (D2Rs) revealed inconsistent abnormalities including asymmetrical bilateral decreases (case 1), unilateral decrease (case 2), and bilateral increases (case 3) in striatal uptakes. CONCLUSION: Data have suggested that presynaptic dopaminergic neurons in JE patients are more susceptible to JE virus than postsynaptic striatal neurons. The degree of movement impairment was more closely correlated to the degree of D2Rs disruption seen in (123)I-IBZM SPECT imaging. PMID- 19548037 TI - Biliary colic preceding acute gallstone pancreatitis. PMID- 19548035 TI - Bioluminescence imaging in mouse models quantifies beta cell mass in the pancreas and after islet transplantation. AB - PURPOSE: We developed a mouse model that enables non-invasive assessment of changes in beta cell mass. PROCEDURES: We generated a transgenic mouse expressing luciferase under control of the mouse insulin I promoter [mouse insulin promoter luciferase-Vanderbilt University (MIP-Luc-VU)] and characterized this model in mice with increased or decreased beta cell mass and after islet transplantation. RESULTS: Streptozotocin-induced, diabetic MIP-Luc-VU mice had a progressive decline in bioluminescence that correlated with a decrease in beta cell mass. MIP Luc-VU animals fed a high-fat diet displayed a progressive increase in bioluminescence that reflected an increase in beta cell mass. MIP-Luc-VU islets transplanted beneath the renal capsule or into the liver emitted bioluminescence proportional to the number of islets transplanted and could be imaged for more than a year. CONCLUSIONS: Bioluminescence in the MIP-Luc-VU mouse model is proportional to beta cell mass in the setting of increased and decreased beta cell mass and after transplantation. PMID- 19548040 TI - Autonomic nerve preservation during rectal cancer resection. PMID- 19548039 TI - A reduction in delayed gastric emptying by classic pancreaticoduodenectomy with an antecolic gastrojejunal anastomosis and a retrogastric omental patch. AB - BACKGROUND: Delayed gastric emptying (DGE) continues to be a major cause of morbidity following pancreaticoduodenectomy (PD). A change in the method of reconstruction following PD was instituted in an attempt to reduce the incidence DGE. METHODS: Patients undergoing PD from January 2002 to December 2008 were reviewed and outcomes determined. Pylorus-preserving pancreaticoduodenectomy (PPPD) with a retrocolic duodenojejunal anastomosis (n = 79) or a classic PD with a retrocolic gastrojejunostomy (n = 36) was performed prior to January 2008. Thereafter, a classic PD with an antecolic gastrojejunal anastomosis and placement of a retrogastric vascular omental patch was undertaken (n = 36). RESULTS: A statistically significant decrease in DGE was noted in the antecolic group compared to the entire retrocolic group (14% vs 40%; p = 0.004) and compared to patients treated by classic PD with a retrocolic anastomosis alone (14% vs 39%; p = 0.016). On multivariate analysis, the only modifiable factor associated with reduced DGE was the antecolic technique with an omental patch, odds ratio (OR) 0.3 (confidence interval (CI) 0.1-0.8) p = 0.022. Male gender was associated with an increased risk of DGE with OR 2.3 (CI 1.1-4.8) p = 0.026. CONCLUSION: A classic PD combined with an antecolic anastomosis and retrogastric vascular omental patch results in a significant reduction in DGE. PMID- 19548038 TI - Small pancreatic and periampullary neuroendocrine tumors: resect or enucleate? AB - OBJECTIVE: The aim of this study was to compare the outcomes of enucleation versus resection in patients with small pancreatic, ampullary, and duodenal neuroendocrine tumors (NETs). METHODS: Multi-institutional retrospective review identified all patients with pancreatic and peri-pancreatic NETs who underwent surgery from January 1990 to October 2008. Patients with tumors < or =3 cm and without nodal or metastatic disease were included. RESULTS: Of the 271 patients identified, 122 (45%) met the inclusion criteria and had either an enucleation (n = 37) and/or a resection (n = 87). Enucleated tumors were more likely to be in the pancreatic head (P = 0.003) or functioning (P < 0.0001) and, when applicable, less likely to result in splenectomy (P = 0.0003). The rate of pancreatic fistula formation was higher after enucleation (P < 0.01), but the fistula severity tended to be worse following resection (P = 0.07). The enucleation and resection patients had similar operative times, blood loss, overall morbidity, mortality, hospital stay, and 5-year survival. However, for pancreatic head tumors, enucleation resulted in decreased blood loss, operative time, and length of stay compared to pancreaticoduodenectomy (P < 0.05). CONCLUSION: These data suggest that most outcomes of enucleation and resection for small pancreatic and peri pancreatic NETs are comparable. However, enucleation has better outcomes than pancreaticoduodenectomy for head lesions and the advantage of preserving splenic function for tail lesions. PMID- 19548041 TI - Prometheus payment model: application to hip and knee replacement surgery. AB - The Prometheus Payment Model offers a potential solution to the failings of the current fee-for-service system and various forms of capitation. At the core of the Prometheus model are evidence-informed case rates (ECRs), which include a bundle of typical services that are informed by evidence and/or expert opinion as well as empirical data analysis, payment based on the severity of patients, and allowances for potentially avoidable complications (PACs) and other provider specific variations in payer costs. We outline the methods and findings of the hip and knee arthroplasty ECRs with an emphasis on PACs. Of the 2076 commercially insured patients undergoing hip arthroplasty in our study, PAC costs totaled $7.8 million (14% of total costs; n = 699 index PAC stays). Similarly, PAC costs were $12.7 million (14% of total costs; n = 897 index PAC stays) for 3403 patients undergoing knee arthroplasty. By holding the providers clinically and financially responsible for PACs, and by segmenting and quantifying the type of PACs generated during and after the procedure, the Prometheus model creates an opportunity for providers to focus on the reduction of PACs, including readmissions, making the data actionable and turn the waste related to PAC costs into potential savings. PMID- 19548042 TI - The Mark Coventry Award: Articular contact estimation in TKA using in vivo kinematics and finite element analysis. AB - In vivo fluoroscopy is a well-known technique to analyze joint kinematics of the replaced knee. With this method, however, the contact areas between femoral and tibial components, fundamental for monitoring wear and validating design concepts, are hard to identify. We developed and tested a novel technique to assess condylar and post-cam contacts in TKA. The technique uses in vivo motion data of the replaced knee from standard fluoroscopy as input for finite element models of the prosthesis components. In these models, tibiofemoral contact patterns at the condyles and post-cam articulations were calculated during various activities. To test for feasibility, the technique was applied to a bicruciate posterior-stabilized prosthesis. Sensitivity of the finite element analysis, validation of the technique, and in vivo tests were performed. To test for potential in the clinical setting, five patients were preliminarily analyzed during chair rising-sitting, stair climbing, and step up-down. For each task and patient, the condylar contact points and contact line rotation were calculated. The results were repeatable and consistent with corresponding calculations from traditional fluoroscopic analysis. Specifically, natural knee kinematics, which shows rolling back and screw home, seemed replicated in all motor tasks. Post-cam contact was observed on both the anterior and posterior faces. Anterior contact is limited to flexion angle close to extension; posterior contact occurs in deeper flexion but is dependent on the motor task. The data suggest the proposed technique provides reliable information to analyze post-cam contacts. PMID- 19548044 TI - Weight bias and weight loss treatment outcomes in treatment-seeking adults. AB - BACKGROUND: Few studies have explored the relationship between weight bias and weight loss treatment outcomes. PURPOSE: This investigation examined the relationship between implicit and explicit weight bias and (a) program attrition, (b) weight loss, (c) self-monitoring adherence, (d) daily exercise levels and overall caloric expenditure, (e) daily caloric intake, and (f) daily caloric deficit among overweight/obese treatment-seeking adults. METHODS: Forty-six overweight/obese adults (body mass index > or = 27 kg/m(2)) participating in an 18-week, stepped-care, behavioral weight loss program completed implicit and explicit measures of weight bias. Participants were instructed to self-monitor and electronically report daily energy intake, exercise, and energy expenditure. RESULTS: Greater weight bias was associated with inconsistent self-monitoring, greater caloric intake, lower energy expenditure and exercise, creation of a smaller caloric deficit, higher program attrition, as well as less weight loss during the self-help phase of the stepped-care treatment. CONCLUSIONS: Weight bias may interfere with overweight/obese treatment-seeking adults' ability to achieve optimal health. PMID- 19548043 TI - Modified rerouting procedure for failed peroneal tendon dislocation surgery. AB - Recurrent dislocation of the peroneal tendons following operative treatment is relatively uncommon, but can be difficult to treat. We asked whether subligamental transposition of the peroneus brevis tendon, fibular grooving, and reattachment of the superior peroneal retinaculum for failed peroneal tendon dislocation surgery would achieve a stable fixation of the peroneal tendons and whether there would be restrictions of ROM or instability of the hindfoot. We reviewed six female patients (mean age, 24.5 years) with general laxity of joints preoperatively and at 6 weeks and 3, 6, and 12 months postoperatively. Within 1 year postoperatively no recurrence was found. In two ankles the extension was restricted 5 degrees to 10 degrees . In another pronation and supination was restricted 5 degrees each. Stability of the ankle increased in four patients and stayed unchanged in two. AOFAS score increased from a mean value of 36 +/- 20.6 preoperatively to 90 +/- 7 postoperatively at 1 year. We conclude transposition of the peroneus brevis tendon is a reasonable treatment for failed peroneal tendon dislocation surgery. LEVEL OF EVIDENCE: Level IV, therapeutic study (prospective case series). See Guidelines for Authors for a complete description of levels of evidence. PMID- 19548045 TI - Topography of Purkinje cells and other calbindin-immunoreactive cells within adult and hatchling turtle cerebellum. AB - The turtle's cerebellum (Cb) is an unfoliated sheet, so the topography of its entire cortex can be easily studied physiologically by optical recordings. However, unlike the mammalian Cb, little is known about the topography of turtle Purkinje cells (PCs). Here, topography was examined using calbindin-D(28K) immunohistochemistry of adult and hatchling turtles (Trachemys scripta elegans, 2.5-15 cm carapace length). Each Cb was flattened between two Sylgard sheets and fixed in paraformaldehyde. Sections (52 microm thick) were cut parallel to the flattened cortex (tangential), resulting in calbindin-immunolabeled PCs being localized to three to six sections for each turtle. PC position and size were quantified using Neurolucida Image Analysis system. Although hatchling Cb were medial-laterally narrower (3.0 vs. 6.5 mm) and rostral-caudally shorter (2.5 vs. 5.5 mm) than adult Cb, both averaged near 15,000 PCs distributed uniformly. Hatchling PCs were smaller than adult PCs (178 vs. 551 microm(2)) and more densely packed (2,180 vs. 625 cells/mm(2)). Calbindin immunoreactivity also labeled non-PCs along the Cb's marginal rim and its caudal pole. Many of these were very small (22.9 microm(2)) ovoid-shaped cells clustered together, possibly proliferating external granule layer cells. Other labeled cells were larger and fusiform-shaped (12.6 x 33.4 microm) adjacent to inner granule cells along the marginal rim, suggestive of migrating cells. It is not known whether these are new neurons being generated within the adult and hatchling Cb and if they connect to efferent and afferent paths. Based on these anatomical findings, we suggest that unique physiological features may exist along the rim of the turtle Cb. PMID- 19548046 TI - Role of PACAP in controlling granule cell migration. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP), a neuropeptide, regulates a wide variety of cellular functions, but little is known about its role in neuronal cell migration. Recent studies revealed that PACAP has short term, cortical layer-specific effects on neuronal cell migration. In this article, we focus on the role of PACAP in controlling the migration of cerebellar granule cells. PMID- 19548047 TI - Effectiveness of prolonged fasting 18f-FDG PET-CT in the detection of cardiac sarcoidosis. AB - BACKGROUND: The Japanese Ministry of Health and Welfare guidelines (JMHWG) are currently the standard used to diagnose cardiac sarcoidosis. JMHWG incorporate (67)Gallium scintigraphy as a minor criterion, while fasting (18)fluorine-2 fluoro-2-deoxy-D-glucose (FDG) PET is not included. As there is no published data comparing the accuracy of prolonged fasting FDG PET-CT (PF-PET) and Gallium scintigraphy for detecting active cardiac sarcoidosis, we sought to compare these two modalities. METHODS AND RESULTS: We retrospectively reviewed medical records and nuclear images of 76 patients with suspected cardiac sarcoid who had either PF-PET or Gallium scintigraphy between January 2004 and August 2008. Eleven patients were excluded due to inadequate fasting for PF-PET, incomplete records or diagnosis other than sarcoid. Cardiac catheterizations, electrocardiogram interpretations, echocardiography reports, pathology reports, therapeutic interventions, and follow-up findings were correlated to PF-PET and Gallium scintigraphy results. Nuclear images of all patients including controls were reviewed independently by two experienced nuclear physicians blinded to results. Using JMHWG as reference standard, sensitivity, specificity, and accuracy of PF PET were 85%, 90%, and 86.7% and for Gallium scintigraphy were 15%, 80%, and 42.8%. CONCLUSIONS: Relative to Gallium scintigraphy, PF-PET appears to provide greater accuracy for detecting cardiac sarcoidosis. Our findings also highlight the importance of revising JMHWG to incorporate PF-PET and the importance of adequate prolonged fasting prior to FDG PET imaging. PMID- 19548048 TI - Multicenter investigation comparing a highly efficient half-time stress-only attenuation correction approach against standard rest-stress Tc-99m SPECT imaging. AB - BACKGROUND: New iterative algorithms for scatter compensation (SC), noise suppression, and depth-dependent collimator resolution (RR) can shorten rest and stress SPECT acquisitions by 50% while maintaining quality and accuracy equivalent to conventional scans. Full-time stress-only myocardial perfusion SPECT is accurate and efficient when combined with line-source attenuation correction (LSAC). We investigated the potential for half-time stress-only LSAC SPECT by comparing this to conventional rest/stress SPECT in patients imaged for suspected CAD at three different centers. METHODS: One hundred and ten patients (58% men, 53% exercise) had 64 projection rest/stress Tc-99m ECG-gated SPECT with simultaneous Gd-153 LSAC: 18 had 30 kg . m(-2). Surface landmarks were difficult or impossible to palpate in 38% of the patients. The scan quality on the LP and TM views was adequate or better in 100 and 98% of the patients, respectively. Dural puncture was achieved with one needle insertion attempt and within two needle insertion attempts in 84% and 98% of the patients, respectively. The ultrasound-measured depth to the intrathecal space correlated well with the actual needle insertion depth (concordance correlation coefficient = 0.82, accuracy 0.95, precision 0.86), with a tendency to overestimate the depth by just 2.1 +/- 5.4 mm. CONCLUSIONS: Ultrasound imaging of the lumbar spine provides clinically useful information that can facilitate spinal anesthesia in the older orthopedic patient population. PMID- 19548049 TI - Effects of food intake and anesthetic on cardiac imaging and uptake of BMS747158 02 in comparison with FDG. AB - BACKGROUND: BMS747158-02 is an (18)F-labeled agent being developed for PET myocardial perfusion imaging. This study examined impacts of feeding state and anesthetic on cardiac imaging and uptake of this agent in rats in comparison with (18)F-fluorodeoxyglucose (FDG). METHODS AND RESULTS: Studies were performed in rats either nonfasted or food deprived for 20 hours and anesthetized with either sodium pentobarbital (Pentob) or ketamine and xylazine (Ket/Xyl). Influences of the feeding state and anesthesia were examined by measurement of blood glucose levels, and tissue biodistribution and cardiac imaging of BMS747158-02 and FDG. The blood glucose levels were lower in fasted than nonfasted rats before anesthesia (91 +/- 11 vs 122 +/- 10 mg/dL) and the levels did not significantly change when anesthetized with Pentob. However, the levels increased markedly by 262 +/- 64 mg/dL in nonfasted rats anesthetized with Ket/Xyl. At 60 minutes post injection, the heart uptake of FDG was significantly lower in fasted than nonfasted rats (0.2 +/- 0.1 vs 2.8 +/- 1.5%ID/g). However, the heart uptake of BMS747158-02 did not differ under these conditions (3.3 +/- 0.9 vs 3.6 +/- 0.9%ID/g, respectively). In nonfasted rats, the heart uptake of FDG was markedly lower when anesthetized with Ket/Xyl than with Pentobl (0.2 +/- 0.1 vs 2.8 +/- 1.5%ID/g). In contrast, the heart uptake of BMS747158-02 was similar with both anesthetics (3.6 +/- 0.5 vs 3.6 +/- 0.9%ID/g). Consistent with the biodistribution studies, the myocardium was not visible following FDG imaging in fasted rats, but clearly seen with BMS747158-02 in both fasted and nonfasted rats anesthetized with either anesthetic. CONCLUSIONS: Unlike FDG, BMS747158-02 cardiac images are clear and not affected by the feeding state and anesthetics. PMID- 19548052 TI - Zinc deficiency reduces neurogenesis accompanied by neuronal apoptosis through caspase-dependent and -independent signaling pathways. AB - Dietary zinc deficiency may affect zinc homeostasis in the brain and lead to reductions of neurogenesis and neuronal survival. However, the mechanisms responsible for the effects of zinc deficiency on hippocampal neurogenesis and neuronal death remain obscure. In the present study, young CD-1 mice were fed with zinc-deficient diet (0.85 ppm) for 5 weeks. The vesicular zinc was reduced at CA1 and CA3 regions of the hippocampus in zinc-deficient mice. The significant decreased zinc ions was associated with a reduction in proliferating cells labeled with bromo-deoxyuridine (BrdU) and immature neurons labeled with doublecortin (DCX) immunoreactivity in the dentate gyrus of the hippocampus. The processes of DCX-positive neurons were shortened, and flexuously went through into the granular cell layer in zinc-deficient hippocampus. There was also a conspicuous increase in the number of TUNEL-positive cells in the hippocampus after zinc-deficient diet treatment. Meanwhile, the apoptosis proteins, including Fas, Fas ligand (FasL), apoptosis inducing factor (AIF), and caspase-3, were significantly activated in zinc-deficient mouse hippocampus. These data suggest that chronic treatment with zinc-deficient diet results in reduction in hippocampal neurogenesis and increases neuronal apoptosis, indicating that zinc deficiency is associated with destroying structural plasticity in the hippocampus. PMID- 19548055 TI - Effects of low-level laser therapy on bone formed after distraction osteogenesis. AB - This study evaluated the effect of low-level laser therapy (LLLT) on the chemical composition, crystallinity and crystalline structure of bone at the site of distraction osteogenesis. Five rabbits were subjected to distraction osteogenesis (latency = 3 days; rate and frequency = 0.7 mm/day for 7 days; consolidation = 10 days), and three were given LLLT with arsenide-gallium-aluminum (AsGaAl; 830 nm, 40 mW): 10 J/cm(2) dose per spot, applied directly to the distraction osteogenesis site during the consolidation stage at 48 h intervals. Samples were harvested at the end of the consolidation stage. X-ray fluorescence and X-ray diffraction were used to analyze chemical composition, crystallinity and crystalline structure of bone at the distraction osteogenesis site. The analysis of chemical composition and calcium (Ca) and phosphorus (P) ratios revealed greater mineralization in the LLLT group. Diffractograms showed that the crystalline structure of the samples was similar to that of hydroxyapatites. Crystallinity percentages were greater in rabbits that were given LLLT. Crystallinity (41.14% to 54.57%) and the chemical composition of the bone at the distraction osteogenesis site were similar to the that of the control group (42.37% to 49.29%). The results showed that LLLT had a positive effect on the biomodulation of newly formed bone. PMID- 19548054 TI - Bactericidal activity of erbium, chromium:yttrium-scandium-gallium-garnet laser in root canals. AB - The aim of this study was to investigate the effectiveness of the erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser by measuring its bactericidal effect inside root canals experimentally colonized with Enterococcus faecalis. We also determined the optimal conditions for the Er,Cr:YSGG laser to achieve the maximal bactericidal effect. An Er,Cr:YSGG Waterlase laser was used, and its antimicrobial effect was compared with that of sodium hypochlorite (NaOCl) at various concentrations as widely used in clinics. This laser emits photons at a wavelength of 2.78 microm. It is a pulsed laser operating at 20 Hz (20 pulses/s). Significant differences between measurements in the different groups (P < 0.05) were observed, depending on time and power used. The use of NaOCl 5% was the most effective procedure, with NaOCl 0.5% being the least effective; however, laser treatment was as effective as NaOCl 5% when applied at 2 W for 60 s. PMID- 19548056 TI - Evaluation of mechanical properties and porcelain bonded strength of nickel chromium dental alloy fabricated by laser rapid forming. AB - The aim was to evaluate the mechanical properties and porcelain bonded strength of nickel-chromium (Ni-Cr) dental alloy fabricated by laser rapid forming (LRF). The tensile properties and porcelain bonded strengths of LRF Ni-Cr dental alloy were evaluated by tensile tests (five specimens per group) and three-point bending tests (ten specimens per group). The same tests for the cast Ni-Cr dental alloy were used as for the control. The microstructure and the bonding interface of the metal substrate to porcelain were analyzed by optical microscopy (OM), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS). The tensile strength of LRF Ni-Cr dental alloy (840 MPa) was superior to that of the cast Ni-Cr alloy (670 MPa), but the ductility had decreased. The porcelain bonded strength of LRF Ni-Cr dental alloy was 44.7 MPa, which was also superior to that of the cast Ni-Cr dental alloy (41.6 MPa). According to analysis by Student's t-test, the porcelain bonded strengths of the Ni-Cr dental alloy presented statistically significant differences between the groups of LRF and cast alloy (P < 0.05), but the porcelain bonded strengths were all above the acceptable value required by the International Organization for Standardization (ISO) standard 9693 (25 MPa). The coefficient of thermal expansion (CTE, mean value of five specimens) of LRF Ni-Cr dental alloy (14 x 10(-6) degrees C(-1)) was higher than that (13.7 x 10(-6) degrees C(-1)) of the cast Ni-Cr dental alloy. Both LRF and cast Ni-Cr dental alloy had positive Deltaalpha, not exceeding the maximum difference (1 x 10(-6) degrees C(-1)) in CTE. SEM and EDS results showed that all the specimens measured for porcelain bonded strength and prepared from the LRF and cast Ni-Cr alloy exhibited a mixed mode of cohesive and adhesive failure. Evaluation of the mechanical properties and porcelain bonded strengths of LRF Ni-Cr alloy revealed that both superior tensile and porcelain bonded properties can be obtained. With improvement of the technique, it is hoped that the LRF Ni-Cr dental alloy could be a good alternative to the conventional cast Ni-Cr dental alloy in the future. PMID- 19548057 TI - Use of the erbium, chromium:yttrium-scandium-gallium-garnet laser on human enamel tissues. Influence of the air-water spray on the laser-tissue interaction: scanning electron microscope evaluations. AB - The study investigated the influence of varying amounts of air/water spray and the energy used by an erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) 2,780 nm laser when treating dental tissues. The morphological effects produced by the laser interaction on healthy human enamel were evaluated by scanning electron microscopy (SEM). The vestibular and lingual surfaces of ten molars were treated with laser at different power settings; each surface was subdivided into cervical, median, and occlusal parts and treated with different proportions of water spray; the series contained 60 tooth portions. Treatment differed in terms of power setting and air/water percentage. All specimens were then subjected to dehydration and metallisation. At SEM evaluation, the classic aspect of laser-treated enamel was visible: grooves, flakes, shelves and sharp edges, indicative of micro-explosion rather than melting. Vaporisation of the tissue created a clear delimitation from surrounding healthy tissue, with partial respect to the prismatic structure of the treated enamel. The aspect of the enamel was rarely type 1 Silverstone but more frequently type 2 or 3, with prismatic structure not respected and/or completely disordered. These morphological differences appeared to be correlated with the inclination of the laser beam aimed at the enamel prisms and with the percentage of air/water used. The laser system analysed showed itself to be effective at removing human dental enamel. The results appeared to be closely correlated with the variation of the percentage of the laser's water-air spray. PMID- 19548059 TI - The eNOS enhancer AVE 9488: a novel cardioprotectant against ischemia reperfusion injury. AB - Nitric oxide (NO) is an important regulator of vascular and myocardial function. Cardiac ischemia/reperfusion injury is reduced in mice overexpressing endothelial NO synthase (eNOS) suggesting cardioprotection by eNOS. Novel pharmacological substances, so called eNOS enhancers, upregulate eNOS expression and thereby increase NO production. We tested the effects of the eNOS enhancer AVE 9488 on cardiac ischemia/reperfusion injury in vivo in mice. After treatment with the eNOS enhancer AVE 9488 (30 mg/kg/day) or placebo for one week mice underwent 30 min of coronary artery ligation and 24 h of reperfusion in vivo. Ischemia reperfusion damage was significantly reduced in mice treated with the eNOS enhancer when compared to placebo treated mice (infarct/area at risk 65.4 +/- 4.1 vs. 36.9 +/- 4.0%, placebo vs. eNOS enhancer, P = 0.0002). The protective effect was blunted in eNOS knockout mice treated with the eNOS enhancer (infarct/area at risk 64.1 +/- 6.2%, eNOS knockout + eNOS enhancer vs. WT + eNOS enhancer, P = ns). Reactive oxygen species were significantly reduced in mice treated with the eNOS enhancer as indicated by significantly lower malondialdehyde-thiobarbituric acid levels (placebo vs. eNOS enhancer, 3.2 +/- 0.5 vs. 0.8 +/- 0.07 micromol/l, P = 0.0003). Thus pharmacological interventions addressed to increase eNOS derived NO production constitute a promising therapeutic approach to prevent myocardial ischemia/reperfusion injury. PMID- 19548060 TI - Juvenile seropositive myasthenia gravis with anti-MuSK antibody after thymectomy. AB - We report the case of a 17.5-year old girl with generalized myasthenia gravis (MG). When she was 13, she started to complain of episodic diplopia, ptosis and mild fatigability of upper and lower extremity muscles. She was diagnosed with MG 3 years later, after exacerbation of her limb muscle weakness. Acetylcholine receptor antibody was positive (2.56 nmol/l, n < 0.4 nmol/l), anti-MuSK antibodies were not tested at that time. She was treated with a thymectomy. After the operation a gradual worsening of her MG was seen; she reported pirydostygmine intolerance. On retesting, acetylcholine receptor antibodies were negative and she was positive for anti-MuSK antibodies. She received immunosuppressive treatment with prednisone and azathioprine with clinical improvement. She is the third reported patient diagnosed with acetylcholine receptor antibody-positive MG, who became MuSK-positive after thymectomy. Recognition of such clinical patterns may be important for decision making in myasthenia gravis patients. PMID- 19548058 TI - Obesity induced-insulin resistance causes endothelial dysfunction without reducing the vascular response to hindlimb ischemia. AB - Impairment of vascular growth is a hallmark of diabetic complications, but the progression and mechanisms are poorly understood. To determine whether obesity and early diabetes impair endothelium-dependent vasodilatation and vascular response to ischemia, microvascular function as well as angiogenic responses to ischemia were assessed in young (C57) and 6-month-old lean mice (old C57), in obese (db-C57) mice, and in mice suffering an early (db-KsJ) and sustained type 2 diabetes (old db-KsJ). Glycemia gradually increased from the db-C57 to the old db KsJ. Early and established type II diabetes significantly reduced the level of insulin that was significantly increased in obese mice. Endothelial function was assessed in isolated resistance arteries while the angiogenic response induced by unilateral hindlimb ischemia was analyzed, after 28 days, with a laser Doppler flowmeter and angiography. Aging (-21%), obesity (-45%), as well as early (-58%) and sustained type II diabetes (-69%) induced a progressive impairment of the endothelium-dependent relaxation of the gracilis artery. Laser Doppler measurements demonstrated that only early and sustained type II diabetes impaired skin blood flow recovery. Vascular collateralization was reduced with aging and severely impaired in older db-KsJ mice, the two strains of mice in which ischemia reduced eNOS expression. These results demonstrate that endothelial dysfunction induced by obesity is insufficient to alter the angiogenic response to ischemia. Furthermore, the development of frank type II diabetes or increasing age is required to impair the vascular response to hindlimb ischemia. We conclude that additional risk factors or severe endothelial dysfunction may be requisite to impede the angiogenic response to ischemia. PMID- 19548061 TI - The effects of recombinant human TSH on bone turnover in patients after thyroidectomy. AB - Thyrotropin receptors are expressed in several extrathyroidal tissues including bone. We investigated whether the increase of thyroid-stimulating hormone (TSH) levels, under stable thyroid hormone levels, affects the bone markers. Thirty-two postmenopausal women, with papillary thyroid carcinoma, previously treated with near-total thyroidectomy and I131 remnant ablation underwent routine evaluation for residual disease by using injections of recombinant human TSH (rhTSH) without withdrawal from thyroxine therapy. Changes in TSH levels and various serum and urine markers of bone metabolism were followed before and 1, 2, 5, and 7 days after the rhTSH injections. A transient, significant decrease in serum calcium and urinary excretion of C- and N-terminal telopeptides of type I collagen was observed after the injections of rhTSH. Serum parathyroid hormone (PTH) started to rise along with TSH, but a significant increase of PTH was only reached on Day 5 when the TSH concentration had fallen more than 80% of the peak value. Bone alkaline phosphatase and osteocalcin did not show any significant change over time. There was no significant correlation between TSH concentration and the various parameters we measured. The study provides evidence that rhTSH produces a transient inhibition of bone resorption, as well as an attenuation of osteoblast response in spite of the PTH activation. Additional studies are needed to resolve the mechanisms by which TSH alters the response of the bone cells. PMID- 19548062 TI - Bone metabolism after cinacalcet administration in patients with secondary hyperparathyroidism. AB - Cinacalcet, an allosteric modulator of a calcium (Ca)-sensing receptor, significantly suppresses parathyroid hormone (PTH) secretion and bone turnover rate in chronic hemodialysis (HD) patients with secondary hyperparathyroidism (SHPT). In this study, bone metabolism after cinacalcet treatment was examined, because hungry bone syndrome is sometimes experienced after parathyroidectomy in severe SHPT. We conducted a prospective observational study in 17 HD patients with SHPT. Cinacalcet was started at 25 mg/day, and the dose was increased step by step based on serum calcium level. A significant decrease in serum Ca and intact PTH concentration was found within 2 weeks. Tartrate-resistant acid phosphatase 5b, a good bone resorption marker, was significantly decreased at week 2 of the study. Serum bone alkaline phosphatase, a marker of bone formation, was increased at week 2 compared with the basal level. It became, however, gradually decreased until week 14. Only one patient whose bone turnover was considerably high had a mild numbness feeling. These results suggest that cinacalcet treatment might transiently accelerate bone formation with rapid suppression of bone resorption. This uncoupling could be involved in a mechanism by which cinacalcet decreases serum Ca level. PMID- 19548064 TI - Is there evidence in support of the use of intra-articular hyaluronate in treating rheumatoid arthritis of the knee? A meta-analysis of the published literature. AB - Intra-articular hyaluronate (HA) injections for treating rheumatoid knee are still debatable, and this meta-analysis aims to elucidate the effectiveness of HA injection for rheumatoid knee. The meta-analysis comprised randomized clinical trials (RCTs) that compared the efficacy of HA injections with that of a placebo. The articles were retrieved after systematic searches of databases, including MEDLINE, EMBASE, and Japana Centra Revuo Medicina. The outcomes were classified into four categories: evaluation of reduction in the intensity of pain, evaluation of reduction in the intensity of inflammation, overall evaluation of therapeutic efficacy, and evaluation of adverse effects. Effect sizes were calculated from the risk ratio (RR) of each of the above-mentioned outcome categories. Five RCTs (720 participants) were pooled for the meta-analysis. The pooled effect sizes were 1.64 (p = 0.01) for pain reduction, 1.61 (p = 0.001) for reduction in inflammation, and 1.50 (p = 0.004) for the overall evaluation of treatment effectiveness. No serious side-effects were reported, while minor adverse effects were reported in patients after HA treatment (RR 0.98, p = 0.32). The results indicated that intra-articular HA is an effective and safe alternative therapy for the rheumatoid knee. PMID- 19548063 TI - Cultivating the uncultured: limits, advances and future challenges. AB - Since the invention of the Petri dish, there have been continuous efforts to improve efficiency in microbial cultivation. These efforts were devoted to the attainment for diverse growth conditions, simulation of in situ conditions and achievement of high-throughput rates. As a result, prokaryotes catalysing novel redox reactions as well as representatives of abundant, but not-yet cultured taxa, were isolated. Significant insights into microbial physiology have been made by studying the small number of prokaryotes already cultured. However, despite these numerous breakthroughs, microbial cultivation is still a low throughput process. The main hindrance to cultivation is likely due to the prevailing lack of knowledge on targeted species. In this review, we focus on the limiting factors surrounding cultivation. We discuss several cultivation obstacles, including the loss of microbial cell-cell communication following species isolation. Future research directions, including the refinement of culture media, strategies based on cell-cell communication and high-throughput innovations, are reviewed. We further propose that a combination of these approaches is urgently required to promote cultivation of uncultured species, thereby dawning a new era in the field. PMID- 19548066 TI - Management of infection in patients with acute leukemia during chemotherapy in Japan: questionnaire analysis by the Japan Adult Leukemia Study Group. AB - Guidelines for the management of febrile neutropenia (FN), deep fungal infection or use of granulocyte colony-stimulating factor (G-CSF) published in the US and Europe cannot be directly applied in other countries. In this study, we undertook a questionnaire survey of member institutions of the Japan Adult Leukemia Study Group to investigate the status of, and problems with, the management of infectious complications in patients with acute leukemia. The questionnaire consisted of 52 multiple-choice questions covering therapeutic environment, antibacterial, and antifungal prophylaxis, empirical therapy (ET) for FN, and use of G-CSF. The results were compared to a previous survey performed in 2001. Usable responses were received from 134 of 184 (71.7%) institutions. With regard to antibacterial prophylaxis, fluoroquinolones and sulfamethoxazole-trimethoprim were most commonly used. Regarding antifungal prophylaxis, the most frequently used agent was fluconazole, followed by itraconazole. In ET for FN, monotherapy with cephems or carbapenems accounted for almost all of the responses. Most respondents indicated that they used micafungin (MCFG) in ET. Prophylactic use of G-CSF during remission induction therapy in acute myeloid leukemia was reported by only 4% of respondents. Strategies for antibacterial and antifungal prophylaxis or treatment of FN should be reviewed and updated as needed. PMID- 19548065 TI - Saponin fraction from Astragalus membranaceus roots protects mice against polymicrobial sepsis induced by cecal ligation and puncture by inhibiting inflammation and upregulating protein C pathway. AB - Sepsis remains the leading cause of death in intensive care units. Uncontrolled systemic inflammation and an impaired protein C pathway are two important contributors to sepsis pathophysiology. Based on the beneficial effects of the saponin fraction from Astragalus membranaceus roots (SAM) against inflammation, liver dysfunction, and endothelium injury, we investigated the potential protective roles and underlying mechanisms of SAM on polymicrobial sepsis induced by cecal ligation and puncture (CLP) in mice. SAM, orally administered 1 h before and after CLP, significantly elevated the survival rate of mice. At 96 h after CLP operation, all mice in the model group died, whereas 33.3% of mice in the SAM (400 mg/kg)-treated group survived. SAM attenuated both inflammatory factors and their abilities to induce tissue dysfunction, which was mainly evidenced by decreased infiltration of polymorphonuclear leukocytes, tissue edema, and lung wet-to-dry weight ratio, lowered levels of myeloperoxidase (MPO), nitric oxide (NO), lactate dehydrogenase (LDH), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in serum, as well as downregulated expressions of iNOS and IL-1beta mRNA in livers. Furthermore, we addressed the effects of SAM on the protein C (PC) pathway, closely linked with sepsis. In CLP-induced septic mice, SAM elevated the impaired expression of PC mRNA in livers. In vitro, SAM reversed the decreased expressions of thrombomodulin (TM) and endothelial PC receptor (EPCR) mRNA induced by lipopolysaccharide (LPS) in endothelial cells. These findings suggest that SAM is able to restore the impaired protein C pathway. Taken together, the current study demonstrates that SAM has protective effects on polymicrobial sepsis in mice. The mechanisms of action involve anti-inflammation and upregulation of the PC pathway. PMID- 19548067 TI - Trends in the seroprevalence of HTLV-1 in Japanese blood donors in Nagasaki Prefecture, 2000-2006. AB - Human T cell leukemia virus type-1 (HTLV-1) is the established cause of adult T cell leukemia/lymphoma. Monitoring time trends in HTLV-1 seroprevalence in blood donors is important to assess the safety of the blood supply in the viral endemic area. We analyzed changes in HTLV-1 seroprevalence in 48415 first-time blood donors who donated blood from 2000 to 2006 in Nagasaki prefecture, an endemic area in Japan. The donors were divided into 10-year birth cohorts: before 1950, 1951-1960, 1961-1970, 1971-1980, and 1981-1990. Among the first-time blood donors, 622 were tested positive for HTLV-1 [overall seroprevalence: 1.28%, (95% CI 1.19-1.39)]. Seroprevalence was significantly high in the birth cohort of before 1950 (6.22%) and declined with birth-year. The time trend of the birth cohort-specific seroprevalence showed almost no change within each birth cohort, except for the birth cohort of 1981-1990 that showed a significantly declining trend (P for trend = 0.006). Among the birth cohort of 1981-1990, the seroprevalence was stable among those born during 1981-1986 (0.66-0.83%), but was lower among those born during 1987-1990 (0-0.38%). Detail analyses showed that HTLV-1 seroprevalence among blood donors clearly declined in those born after 1987. PMID- 19548068 TI - Differential diagnosis and treatment of primary, cutaneous, anaplastic large cell lymphoma: not always an easy task. AB - Primary cutaneous anaplastic large cell lymphoma (PC-ALCL) is a rare and distinct neoplasm appearing de novo on the skin. We present a case of a 75-year-old man diagnosed with PC-ALCL in his left femoral region. We describe the morphology of lesions along with the differential diagnosis, treatment, clinical course and prognosis. We further discuss parameters concerning treatment that should be considered when a PC-ALCL is diagnosed. Our case report demonstrates the complexity in classification, staging, differential diagnosis and therapy selection of PC-ALCLs. It is crucial to emphasize the importance of clinical criteria in diagnosing a PC-ALCL in combination with immunohistochemistry. PMID- 19548070 TI - Prospective assessment of quality of life in adult patients with primary brain tumors in routine neurooncology practice. AB - The aim of this article is to evaluate and assess the impact of various factors on quality of life (QOL) in adult patients with primary brain tumors seen consecutively in routine neurooncology practice. Two hundred and fifty-seven adult patients, after undergoing surgical intervention and histologically proven primary brain neoplasms were registered in the NeuroOncology Clinic at our centre during 1 full calendar year. The study included detailed neurological assessment, evaluation of QOL using EORTC questionnaire (QLQ-30) and specific Brain Cancer module (BN 20). In the present analysis, QOL scores before starting adjuvant treatment were measured and impact of patient and tumor related factors were analyzed. Baseline global QOL data of all patients (available in 243) was relatively low including in all histological tumor types. Physical function, role function, emotion function, cognitive and social function scores were 80, 78, 65.7, 70 and 70.5 (higher values better), respectively. Domains of future uncertainty, visual disorder, motor deficit, communication deficit, headache, seizures and drowsiness scores were 19.6, 18.2, 28.5, 30.7, 21, 31.8 and 16 (lower values better), respectively. Elderly patients had poorer global score (21 points difference; p = 0.161). Patients with lower performance status (KPS < 70) had a lower global QOL (KPS >or= 80 vs. or=2 days and no apparent contraindications for thromboprophylaxis were grouped into the categories of critical care, surgery and medically ill before being assessed for additional VTE risk factors based on the diagnostic criteria outlined in ACCP7. For patients at risk, the recommended type (mechanical or pharmacologic), dose, and duration of thromboprophylaxis was identified based on the guidelines and compared to the regimen actually received, if any. Among the 258,556 hospitalized patients, 68,278 (26.4%) were determined to be at risk of VTE without apparent contraindications for thromboprophylaxis. The proportions of patients who received the appropriate type, dose, and duration of thromboprophylaxis were 10.5, 9.8, and 17.9% for critical care, medical, and surgical patients, respectively. Of those at risk, 36.8% received no thromboprophylaxis and an additional 50.2% received thromboprophylaxis deemed inappropriate for one or more reasons. The implementation of ACCP7 guidelines for type, dosage, and duration of thromboprophylaxis is low in patients at risk of VTE. There is a need for physicians and health systems to improve awareness and implementation of recommended thromboprophylaxis. PMID- 19548073 TI - The synthesis of magnetic and fluorescent bi-functional silica composite nanoparticles via reverse microemulsion method. AB - In this paper, a simple synthesis method of small-size (about 50 nm in diameter), high magnetic and fluorescent bi-functional silica composite nanoparticles were developed, in which water-soluble Fe3O4 magnetic nanoparticles (MNs) and CdTe quantum dots (QDs) were directly incorporated into a silica shell by reverse microemulsion method. The high luminescent QDs can be used as luminescent marker, while the high magnetic MNs allow the manipulation of the bi-functional silica composite nanoparticles by external magnetic field. Poly (dimethyldiallyl ammonium chloride) was used to balance the electrostatic repulsion between CdTe QDs and silica intermediates to enhance the fluorescence intensity of MNs QDs/SiO2 composite nanoparticles. The optical property, magnetic property, size characterization of the bi-functional composite nanoparticles were studied by UV Vis and PL emission spectra, VSM, TEM, SEM. The stabilities toward time, pH and ionic strength and the effect of MNs on the fluorescence properties of bi functional silica composite nanoparticles were also studied in detail. By modifying the surface of MNs-QDs/SiO2 composite nanoparticles with amino and methylphosphonate groups, biologically functionalized and monodisperse MNs QDs/SiO2 composite nanoparticles can be obtained. In this work, bi-functional composite nanoparticles were conjugated with FITC labeled goat anti-rabbit IgG, to generate novel fluorescent-magnetic-biotargeting tri-functional composite nanoparticles, which can be used in a number of biomedical application. PMID- 19548074 TI - Lipopolysaccharide induced upregulation of beta-1,4-galactosyltransferase-I in Schwann cell. AB - beta4 Galactosylation of glycoproteins is one of the most important post translational modifications. Recent studies have demonstrated that aberrant galactosylation associates with some inflammation diseases. beta-1,4 galactosyltransferase-I (beta-1,4-GalT-I), which transfers galactose to the terminal N-acetylglucosamine of N- and O-linked glycans in a beta-1,4- linkage, considered to be the major galactosyltransferse among the seven members of the subfamily responsible for beta4 galactosylation. In the present study, we investigated the expression of beta-1,4-GalT-I in Schwann cells under Lipopolysaccharide (LPS) treatment. RT-PCR revealed that the beta-1,4-GalT-I mRNA was significant increased as early as 2 h after LPS stimulation. Immunofluorescence showed that beta-1,4-GalT-I was located in Golgi apparatus and membrane of Schwann cells. With the 1 microg/ml LPS treatment, expression levels of beta-1,4-GalT-I was much higher compared with control group. In addition, lectin blot indicated that the beta4 galactosylation of glycoproteins such as integrin alpha5 was enhanced, which may due to the induced beta-1,4-GalT-I expression. These results suggested that beta-1,4-GalT-I may play an important role in adhesion and migration of Schwann cells during inflammation. PMID- 19548075 TI - RS virus-induced inflammation and the intracellular glutathione redox state in cultured human airway epithelial cells. AB - There is ample evidence that asthma is mediated by oxidative stress and that viral infection, which is associated with asthma onset and exacerbation in infants, acts as one type of oxidative stress. The goal of this study was to determine whether respiratory syncytial virus (RSV) induces oxidative stress in cultured A549 human airway epithelial cells and normal human bronchial epithelial cells (NHBE), and whether such RSV-induced oxidative stress can induce airway inflammation. To evaluate the direct effect of RSV infection as an oxidative stressor, the intracellular levels of reduced glutathione (GSH) or oxidized glutathione (GSSG) were measured. Their ratio (GSH/GSSG) was calculated to indicate intracellular oxidation-reduction (redox) status in A549 and NHBE. To evaluate the extent to which glutathione redox regulation affected cytokine/chemokine production, the effect of pretreatment with a reductive agent, glutathione monoethyl ester (GSH-OEt) and RSV-specific monoclonal antibody was thus studied. RSV acted as a potent oxidative stressor on the intracellular glutathione redox state in human airway epithelial cells, activating signals to increase the production of cytokine/chemokine. Pretreatment with GSH-OEt significantly suppressed RSV-induced time-dependent changes in the intracellular redox state, and also suppressed RSV-induced up-regulation of epithelial cell derived IL-8, IL-6 and eotaxin production, as well as RSV-specific monoclonal antibody. RSV-induced oxidative stress is likely to contribute to the perpetuation and amplification of the inflammatory response. Therapeutic intervention against oxidative stress might therefore be beneficial as adjunctive therapies for respiratory disorders that are caused by an RSV infection. PMID- 19548076 TI - Pharmacological treatment of patients with chronic critical limb ischemia: L propionyl-carnitine enhances the short-term effects of PGE-1. AB - PURPOSE: To evaluate the therapeutic effects of L-propionyl-carnitine (LPC) in patients with critical limb ischemia (CLI), as defined by the TASC guidelines. METHODS: The study, double-blinded, randomised, assessed intravenous infusion of LPC 1.2 g/day in combination with PGE-1, 60 mg/day (LPC group: 37 patients), or PGE-1 only (control group: 38 patients) in a total of 75 patients suffering from CLI. Treatment duration was 20 days. We evaluated rest pain, maximum walking distance (MWD) and skin ulcer size. RESULTS: In both groups we observed a significant reduction in pain score and ulcer size and an increase in MWD. In the patients treated with the combination, the improvement was greater: median value for pain score decreased from 2.75 to 0.85 in the LPC group and from 2.51 to 1.71 in the control group; MWD increased from 55 M to 130 M in the LPC group, and from 55 M to 102 M in the control group; median decrease of ulcer size was significantly greater in patients treated with LPC + PGE1. CONCLUSIONS: Our study shows that LPC, whose effectiveness on claudication is already known, has favourable effects in patients with CLI, since it reinforces the effects produced by PGE-1. PMID- 19548078 TI - Qualitative or quantitative differences between Asperger's disorder and autism? Historical considerations. AB - The histories of autism and Asperger's Disorder (AD), based on original contributions by Kanner and Asperger, are reviewed in relation to DSM-IV diagnostic criteria. Their original articles appear to have influenced the distinction between AD and autism made in the DSM-IV. Based on up-to-date empirical research, however, it appears that AD and autism are not qualitatively distinct disorders, but are different quantitative manifestations of the same disorder. The differences between AD and autism may be a function of individual variability in these areas, not the manifestation of qualitatively distinct disorders. The DSM-IV criteria for AD and autism need to be considered with their historical developments, and based on empirical evidence, the DSM-IV diagnostic criteria may be subject to critical review. PMID- 19548079 TI - Follicle curetting at the time of oocyte retrieval increases the oocyte yield. AB - PURPOSE: To determine whether follicle curetting at the time of oocyte retrieval increases oocyte yield. METHODS: Retrospective review of all patients who underwent oocyte retrieval from July 1, 2003 to June 30, 2005. MAIN OUTCOME MEASURE: Number of oocytes retrieved. SECONDARY OUTCOME MEASURES: retrieval time, number of cryopreserved embryos, pregnancy rates, and incidence of ovarian hyperstimulation syndrome. RESULTS: There were no differences in patient demographics, antral follicle count, cycle stimulation characteristics, fertilization rates, embryo quantity or quality, embryo cryopreservation rates, clinical pregnancy rates, live birth rates, or ovarian hyperstimulation syndrome between the groups. Retrievals that utilized curetting took three minutes longer. Follicle curetting significantly increased the number of oocytes retrieved, 13.9 +/- 0.6 compared to 11.4 +/- 0.6 oocytes without curetting (P = 0.003). The quantity of mature oocytes was also increased with curetting (10.3 +/- 0.5 versus 8.4 +/- 0.5, P = 0.006). CONCLUSIONS: This study demonstrated that follicle curetting significantly increased oocyte yield. While it did not increase live birth rates, this increase in oocyte yield should lead to increased numbers of embryos for selection at transfer and increased embryos for cryopreservation. PMID- 19548077 TI - A large-scale model of the locust antennal lobe. AB - The antennal lobe (AL) is the primary structure within the locust's brain that receives information from olfactory receptor neurons (ORNs) within the antennae. Different odors activate distinct subsets of ORNs, implying that neuronal signals at the level of the antennae encode odors combinatorially. Within the AL, however, different odors produce signals with long-lasting dynamic transients carried by overlapping neural ensembles, suggesting a more complex coding scheme. In this work we use a large-scale point neuron model of the locust AL to investigate this shift in stimulus encoding and potential consequences for odor discrimination. Consistent with experiment, our model produces stimulus sensitive, dynamically evolving populations of active AL neurons. Our model relies critically on the persistence time-scale associated with ORN input to the AL, sparse connectivity among projection neurons, and a synaptic slow inhibitory mechanism. Collectively, these architectural features can generate network odor representations of considerably higher dimension than would be generated by a direct feed-forward representation of stimulus space. PMID- 19548080 TI - Does the genetic and familial background of males undertaking ICSI affect the outcome? AB - PURPOSE: To investigate whether the success rate of ICSI is (1) related to the etiology of infertility or (2) adversely affected by a family history of potential genetic disorders. METHODS: All men with an ICSI indication in our hospital between 1994 and 2005 were included in our cohort study. Data on the ICSI process, etiology of infertility, and family history were collected. ICSI success rates of infertility subgroups and a subgroup with a positive family history were compared to a group with unknown etiology and a negative family history. RESULTS: There was no significant difference in clinical pregnancy or delivery rates between the subgroups. Couples achieving a pregnancy underwent significantly more ICSI cycles compared to couples not achieving a pregnancy. CONCLUSION: Our results suggest that the success rate of ICSI treatment is not related to the cause of infertility or a family history positive for potential genetic disorders. PMID- 19548081 TI - Follicular and luteal phase endometrial thickness and echogenic pattern and pregnancy outcome in oocyte donation cycles. AB - PURPOSE: To study the effect of endometrial thickness (ET) and echogenic pattern (EP) in oocyte donation cycles upon pregnancy outcomes. METHODS: Seventy-nine cycles resulting in blastocyst embryo transfer were evaluated. Donors underwent ovarian hyperstimulation using rFSH and GnRH-antagonist. Recipients were synchronized to donors using GnRH-agonist down-regulation followed by fixed dose of estrogen (E2) and progesterone (P4) following hCG. Transvaginal ultrasound (US) obtained ET and EP 10-11 days after initiation of E2 and on day of embryo transfer. Primary outcome was ET and EP in pregnant and non-pregnant cycles. Stimulation and embryology data was analyzed in donors to assess differences prior to transfer. RESULTS: Fifty-nine cycles resulted in clinical pregnancy. No differences were observed in pregnant vs. non-pregnant cycles in proliferative or secretory ET and EP. Similar baseline and stimulation characteristics were found in pregnant and non-pregnant cycles. Regression analysis showed end thickness were not predictive of pregnancy outcomes. CONCLUSIONS: Endometrial characteristics in recipients prior to and following progesterone were not predictive of pregnancy outcomes. PMID- 19548082 TI - Molecular evolution of TEPP protein genes in metazoans. AB - TEPP is a gene expressed in human reproductive organs such as testis, prostate, and placenta. Here, identification and molecular evolutionary analysis of TEPP proteins in various metazoan animals including deuterostomes (chordates, hemichordates, and echinoderms), lophotrochozoans (mollusks and annelids), and cnidarians (sea anemone and coral) are reported. A multiple sequence alignment revealed two highly conserved regions in TEPP proteins that had no similarity to any other known domains or proteins. Genomic sequence analysis showed frequent shifting of the splice sites of intron 1 in mammalian TEPP genes. A comparison of the intron positions in the coding region showed that the exon/intron structure of the TEPP gene was established in an early metazoan ancestor and that independent loss of a single intron occurred in echinoderms and in vertebrates. The urochordate tunicate TEPP genes are intronless, possibly due to replacement of the original gene by a retrogene. No homolog was detected in birds, insects, nematodes, and teleost fishes despite the extensive sequence data of these species, implying that the TEPP gene might be lost in these lineages. PMID- 19548083 TI - Inhibition of pulmonary metastasis in a human MT3 breast cancer xenograft model by dual liposomes preventing intravasal fibrin clot formation. AB - The process of metastasis formation in cancer is not completely understood and is the main reason cancer therapies fail. Previously, we showed that dual liposomes simultaneously containing the hemostatic inhibitor, dipyridamole and the anticancer drug, perifosine potently inhibited metastasis, causing a 90% reduction in the number of lung metastases in a murine experimental metastasis model. To gain deeper insight into the mechanisms leading to the inhibition of metastasis by these dual liposomes, in the present study, the development of metastases by MT3 breast cancer cells in a mouse xenograft model was analyzed in more detail with regard to tumor cell settlement and metastatic growth. We found that the development of lung metastases by MT3 tumor cells is essentially dependent on the formation of fibrin clots as a precondition for the pulmonary arrest of tumor cells and the subsequent intravascular expansion of micrometastases before their invasion into the surrounding tissue. PMID- 19548084 TI - Urban-rural differences in breast cancer incidence by hormone receptor status across 6 years in Egypt. AB - Breast cancer incidence is higher in developed countries with higher rates of estrogen receptor positive (ER+) tumors. ER+ tumors are caused by estrogenic exposures although known exposures explain approximately 50% of breast cancer risk. Unknown risk factors causing high breast cancer incidence exist that are estrogenic and development-related. Xenoestrogens are such risk factors but are difficult to study since developed countries lack unexposed populations. Developing countries have urban-rural populations with differential exposure to xenoestrogens. This study assessed urban-rural breast cancer incidence classified by hormone receptor status using data from Gharbiah population-based cancer registry in Egypt from 2001 to 2006. Urban ER+ incidence rate (per 100,000 women) was 2-4 times (IRR = 3.36, 95% CI = 4.84, 2.34) higher than rural incidence rate. ER-incidence rate was 2-3 times (IRR = 1.86, 95% CI = 2.38, 1.45) higher in urban areas than in rural areas. Our findings indicate that urban women may probably have a higher exposure to xenoestrogens. PMID- 19548086 TI - Intermittent hypercapnic hypoxia induced protein changes in the piglet hippocampus identified by MALDI-TOF-MS. AB - Intermittent hypercapnic hypoxia (IHH) induces protein changes in the brainstem, but its effects on the hippocampus have not yet been studied. Using a proteomics based approach, we tested the hypothesis that IHH up-regulates apoptotic promoters and down-regulates apoptotic inhibitors in the developing hippocampus. Male piglets aged 13-14 days were assigned to control (n = 6) or IHH (n = 5) groups. Using two-dimensional polyacrylamide gel electrophoresis, matrix-assisted laser desorption/ionisation-time of flight-mass spectrometry (MALDI-TOF-MS), a total of 26 protein spots were differentially expressed in IHH compared to control group. Thirteen of these (6 up-regulated, 7 down-regulated) were identified including 14-3-3theta/tau (increased), glial fibrillary acidic protein (increased) and a-internexin (decreased). Further analysis with western blot validated these proteins and immunohistochemistry showed specific regional changes in the subiculum, stratum radiatum and CA1 of the hippocampus. Most proteins identified were involved in promoting cell survival under apoptotic conditions. These findings improve our understanding of the cellular processes that occur in the hippocampus during IHH exposure, and have important implications in clinical settings where IHH is experienced, for example, during prone sleeping or with obstructive sleep apnea in an infant. PMID- 19548087 TI - Inhibition of MEK signaling enhances the ability of cytarabine to induce growth arrest and apoptosis of acute myelogenous leukemia cells. AB - The mitogen-activated protein kinase/ERK kinase (MEK)/ERK pathway was shown to be constitutively activated in a large number of acute myelogenous leukemia (AML) cells, suggesting the important roles of this pro-survival signaling in leukemogenesis and proliferation of AML cells. This study explored the impact of the MEK inhibitor AZD6244 on the effect of cytarabien (AraC), one of the most commonly used anti-leukemia agents, to induce growth arrest and apoptosis of AML cells. AZD6244 effectively blocked AraC-induced MEK/ERK activation and enhanced its ability to induce growth arrest and apoptosis of NB4 and HL60 cells in parallel with induction of DNA damage as measured by detection of gamma-H2AX by Western Blot analysis, resulting in enhanced expression of p21( waf1 ) and downregulation of c-Myc and Bcl-xl in these cells. Enhanced induction of apoptosis mediated by combination of AZD6244 and AraC was also shown in freshly isolated AML cells (n = 3). Taken together, concomitant administration of AraC and the inhibitor of MEK/ERK signaling may be useful for treatment of individuals with AML. PMID- 19548085 TI - Killing of Bacillus spores is mediated by nitric oxide and nitric oxide synthase during glycoconjugate-enhanced phagocytosis. AB - Nitric oxide (NO) is a signaling and defense molecule of major importance. NO endows macrophages with bactericidal, cytostatic as well as cytotoxic activity against various pathogens. Bacillus spores can produce serious diseases, which might be attenuated if macrophages were able to kill the spores on contact. Present research was carried out to study whether glycoconjugates stimulated NO and nitric oxide synthase (NOS2) production during phagocytosis killing of Bacillus spores. Murine macrophages exposed to glycoconjugate-treated spores induced NOS2 and NO production that was correlated with high viability of macrophages and killing rate of bacterial spores. Increased levels of inducible NOS2 and NO production by macrophages in presence of glycoconjugates suggested that the latter provide an activation signal directed to macrophages. Glycoconjugates were shown to exert a protective influence, sparing macrophages from spore-induced cell death. In presence of glycoconjugates, macrophages efficiently kill the organisms. Without glycoconjugate activation, murine macrophages were ineffective at killing Bacillus spores. These results suggest that glycoconjugates promote killing of Bacillus spores by blocking spore-induced macrophage cell death, while increasing their activation level and NO and NOS2 production. Glycoconjugates suggest novel antimicrobial approaches to prevention and treatment of infection caused by bacterial spores. PMID- 19548088 TI - Quantitative analysis of single bacterial chemotaxis using a linear concentration gradient microchannel. AB - A microfluidic device to quantify bacterial chemotaxis has been proposed, which generates a linear concentration gradient of chemoattractant in the main channel only by convective and molecular diffusion, and which enables the bacteria to enter the main channel in a single file by hydrodynamic focusing technique. The trajectory of each bacterium in response to the concentration gradient of chemoattractant is photographed by a CCD camera and its velocity is acquired by a simple PTV (Particle Tracking Velocimetry) algorithm. An advantage of this assay is to measure the velocity of a single bacterium and to quantify the degree of chemotaxis by analyzing the frequency of velocities concurrently. Thus, the parameter characterizing the motility of wild-type Escherichia coli strain RP437 in response to various concentration gradients of L-aspartate is obtained in such a manner that the degree of bacterial chemotaxis is quantified on the basis of a newly proposed Migration Index. PMID- 19548090 TI - Identification of family-specific residue packing motifs and their use for structure-based protein function prediction: II. Case studies and applications. AB - This paper describes several case studies concerning protein function inference from its structure using our novel approach described in the accompanying paper. This approach employs family-specific motifs, i.e. three-dimensional amino acid packing patterns that are statistically prevalent within a protein family. For our case studies we have selected families from the SCOP and EC classifications and analyzed the discriminating power of the motifs in depth. We have devised several benchmarks to compare motifs mined from unweighted topological graph representations of protein structures with those from distance-labeled (weighted) representations, demonstrating the superiority of the latter for function inference in most families. We have tested the robustness of our motif library by inferring the function of new members added to SCOP families, and discriminating between several families that are structurally similar but functionally divergent. Furthermore we have applied our method to predict function for several proteins characterized in structural genomics projects, including orphan structures, and we discuss several selected predictions in depth. Some of our predictions have been corroborated by other computational methods, and some have been validated by independent experimental studies, validating our approach for protein function inference from structure. PMID- 19548089 TI - Multidisciplinary management of primary tumors of the vertebral column. AB - OPINION STATEMENT: Primary spinal neoplasms are rare tumors that can lead to significant morbidity secondary to local bone destruction and invasion into adjacent neurological and vascular structures. These tumors represent a clinical challenge to even the most experienced physicians and require a multidisciplinary approach to ensure optimal patient outcomes. This review will discuss the most common primary bone tumors and focus on recent surgical, medical, and radiation treatment advances. PMID- 19548091 TI - Recoupling and decoupling of nuclear spin interactions at high MAS frequencies: numerical design of CN(n)(nu) symmetry-based RF pulse schemes. AB - The CN(n)(nu) class of RF pulse schemes, commonly employed for recoupling and decoupling of nuclear spin interactions in magic angle spinning solid state NMR studies of biological systems, involves the application of a basic "C" element corresponding to an RF cycle with unity propagator. In this study, the design of CN(n)(nu) symmetry-based RF pulse sequences for achieving 13C-13C double-quantum dipolar recoupling and through bond scalar coupling mediated 13C-13C chemical shift correlation has been examined at high MAS frequencies employing broadband, constant-amplitude, phase-modulated basic "C" elements. The basic elements were implemented as a sandwich of a small number of short pulses of equal duration with each pulse characterised by an RF phase value. The phase-modulation profile of the "C" element was optimised numerically so as to generate efficient RF pulse sequences. The performances of the sequences were evaluated via numerical simulations and experimental measurements and are presented here. PMID- 19548092 TI - TALOS+: a hybrid method for predicting protein backbone torsion angles from NMR chemical shifts. AB - NMR chemical shifts in proteins depend strongly on local structure. The program TALOS establishes an empirical relation between 13C, 15N and 1H chemical shifts and backbone torsion angles phi and psi (Cornilescu et al. J Biomol NMR 13 289 302, 1999). Extension of the original 20-protein database to 200 proteins increased the fraction of residues for which backbone angles could be predicted from 65 to 74%, while reducing the error rate from 3 to 2.5%. Addition of a two layer neural network filter to the database fragment selection process forms the basis for a new program, TALOS+, which further enhances the prediction rate to 88.5%, without increasing the error rate. Excluding the 2.5% of residues for which TALOS+ makes predictions that strongly differ from those observed in the crystalline state, the accuracy of predicted phi and psi angles, equals +/-13 degrees . Large discrepancies between predictions and crystal structures are primarily limited to loop regions, and for the few cases where multiple X-ray structures are available such residues are often found in different states in the different structures. The TALOS+ output includes predictions for individual residues with missing chemical shifts, and the neural network component of the program also predicts secondary structure with good accuracy. PMID- 19548093 TI - Geochemical provenance and spatial distribution of fluoride in groundwater of Mayo Tsanaga River Basin, Far North Region, Cameroon: implications for incidence of fluorosis and optimal consumption dose. AB - The 500,000 inhabitants of Mayo Tsanaga River Basin are vulnerable to a "silent" fluorosis from groundwater consumption. For the first time, the groundwater is investigated for the purpose of identifying the provenance of fluoride and estimating an optimal dose of fluoride in the study area. Based on the fluoride content of groundwater, fluorine and major oxides abundances in rocks from the study area, mean annual atmospheric temperature, and on-site diagnosis of fluorosis in children, the following results and conclusions are obtained: Fluoride concentration in groundwater ranges from 0.19 to 15.2 mg/l. Samples with fluoride content of <1.5 mg/l show Ca-HCO(3) signatures, while those with fluoride >1.5 mg/l show a tendency towards Na-HCO(3) type. Fluor-apatite and micas in the granites were identified as the main provenance of fluoride in the groundwater through water-rock interactions in an alkaline medium. The optimal fluoride dose in drinking water of the study area should be 0.7 mg/l, and could be adjusted downward to a level of 0.6 mg/l due to the high consumption rate of groundwater, especially during drier periods. PMID- 19548095 TI - Forensic entomology: a template for forensic acarology? AB - Insects are used in a variety of ways in forensic science and the developing area of forensic acarology may have a similar range of potential. This short account summarises the main ways in which entomology currently contributes to forensic science and discusses to what extent acarology might also contribute in these areas. PMID- 19548094 TI - Arsenic detoxification potential of aox genes in arsenite-oxidizing bacteria isolated from natural and constructed wetlands in the Republic of Korea. AB - Arsenic is subject to microbial interactions, which support a wide range of biogeochemical transformations of elements in natural environments such as wetlands. The arsenic detoxification potential of the bacterial strains was investigated with the arsenite oxidation gene, aox genotype, which were isolated from the natural and constructed wetlands. The isolates were able to grow in the presence of 10 mM of sodium arsenite (As(III) as NaAsO(2)) and 1 mM of D: +glucose. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that these isolated strains resembled members of the genus that have arsenic-resistant systems (Acinetobacter sp., Aeromonas sp., Agrobacterium sp., Comamonas sp., Enterobacter sp., Pantoea sp., and Pseudomonas sp.) with sequence similarities of 81-98%. One bacterial isolate identified as Pseudomonas stutzeri strain GIST BDan2 (EF429003) showed the activity of arsenite oxidation and existence of aoxB and aoxR gene, which could play an important role in arsenite oxidation to arsenate. This reaction may be considered as arsenic detoxification process. The results of a batch test showed that P. stutzeri GIST-BDan2 (EF429003) completely oxidized in 1 mM of As(III) to As(V) within 25-30 h. In this study, microbial activity was evaluated to provide a better understanding of arsenic biogeochemical cycle in both natural and constructed wetlands, where ecological niches for microorganisms could be different, with a specific focus on arsenic oxidation/reduction and detoxification. PMID- 19548096 TI - Assessment of surface water quality of the Ceyhan River basin, Turkey. AB - In this study, surface water quality of the Ceyhan River basin were assessed and examined with 13 physico-chemical parameters in 31 stations in 3 months during the period of 2005. Multivariate statistical techniques were applied to identify characteristics of the water quality in the studied stations. Nutrients, Cl- and Na+ affected mostly to the stations of Erkenez 2, Sir 2, and Sir 3 in the ordination diagram of correspondence analysis. Three factors were extracted by principal component analysis, which explains 79.14% of the total variation. The first factor (PC1) captures variables of EC, DO, NO2-, PO4triple bond, Cl-, SO4=, Na+, and Ca++. The second factor (PC2) is significantly related to pH, NH3-, and Mg++, while water temperature (T) and NO3- accounted for the greatest loading for factor 3 (PC3). The stations were divided into three groups for PC1, two groups for PC2, and three groups for PC3 by hierarchical cluster analysis. The stations in the vicinity of cities presented low dissolved oxygen and high concentration of physico-chemical parameter levels. The stations of Erkenez 2, Sir 2, Sir 3, and Aksu 4 located near the city of Kahramanmaras were characterized by an extremely high pollution due to discharge of wastewater from industry and domestic. Pinarbasi and Elbistan stations were also influenced by household wastewater of the city of Elbistan. According to criteria of Turkish Water Pollution Control Regulation, Erkenez 2, Sir 2, and Sir 3 stations have high polluted water. This study suggests that it is urgent to control point pollutions, and all wastewater should be purified before discharge to the Ceyhan River basin. PMID- 19548097 TI - Seasonal variation of different microorganisms with nickel and cadmium in the industrial wastewater and agricultural soils. AB - Wastewater and soil samples were collected from the industrial area of Ghaziabad City, India from January 2005 to December 2007 and were analyzed for the presence of heavy metals by atomic absorption spectrophotometry. Test samples revealed high levels of Fe, Cr, Cu, Ni, Zn, and Cd as 967.03, 34.63, 27.97, 19.7, 16.70, and 3.20 mg/L of wastewater, respectively. The concentrations of inorganic minerals were higher in the soil samples irrigated with wastewater. Total coliforms were found to be maximum (1,133x10(4) most probable number per 100 mL) during spring and summer followed by winter and postmonsoon in the wastewater samples. The microbial count in soil as well as in wastewater decreases as the metal concentration increases. The concentration 200 microg/mL of nickel and cadmium inhibits majority of the population, while, at some points, it inhibits 100% of the population. The exponential decay model for microbial count at the increasing metal concentrations indicate that asymbiotic N2 fixers were best fitted to the model. In all the seasons, the order of decline in terms of exponential decay of the population of different microbial groups in soil was asymbiotic N2 fixers>actinomycetes>fungi>aerobic heterotrophic bacteria. The different microbial groups that have different values of slope in different seasons indicate that the resistant population of microorganisms was variable with seasons. PMID- 19548098 TI - Parallel microwave chemistry in silicon carbide reactor platforms: an in-depth investigation into heating characteristics. AB - The heating behavior of silicon carbide reaction platforms under 2.45 GHz microwave irradiation was investigated with the aid of online thermoimaging cameras and multiple-channel fiber-optic probe temperature sensors placed inside the wells/vials of the silicon carbide microtiter plates. Microwave irradiation leads to a rapid and homogeneous heating of the entire plate, with minimal deviations in the temperature recorded at different positions of the plate or inside the wells. In temperature-controlled experiments using dedicated multimode reactors, solvents with different microwave absorption characteristics can be heated in parallel in individual wells/vials of the silicon carbide plate reaching the same set temperature. Due to the large heat capacity and high thermal conductivity of silicon carbide, the plates are able to moderate any field inhomogeneities inside a microwave cavity. Although the heating of the plates can be performed extremely efficiently inside a microwave reactor, heating and synthetic applications can alternatively be carried out by applying conventional conductive heating of the silicon carbide plates on a standard hotplate. Due to the slower heating of the silicon carbide material under these conditions, somewhat longer reaction times will be required. PMID- 19548099 TI - Comparison between some oxidative stress biomarkers values in serum and plasma of clinically healthy adult camels (Camelus dromedarius) in Iran. AB - In this study, we measured some oxidative stress biomarkers, MDA and total thiol groups content in plasma along by this parameters in serum and Ferric Reducing Ability of Plasma (FRAP) of camels. The present study was undertaken first to establish a background of free radical damage status and second to comparison these levels in serum and plasma. To accomplish this, blood samples from 50 male Dromedary clinically healthy adult camels aged 24-48 months were collected. The correlation between serum and plasma levels was calculated for MDA and total thiol groups content. Our results revealed that only lipid peroxides in serum could be useful to predict the oxidative stress in plasma significantly (P<=0.05). PMID- 19548100 TI - Is the German suspension of MON810 maize cultivation scientifically justified? AB - We examined the justifications invoked by the German government in April 2009 to suspend the cultivation of the genetically modified maize varieties containing the Bt insect-resistance trait MON810. We have carried out a critical examination of the alleged new data on a potential environmental impact of these varieties, namely two scientific papers describing laboratory force-feeding trials on ladybirds and daphnia, and previous data on Lepidoptera, aquatic and soil organisms. We demonstrate that the suspension is based on an incomplete list of references, ignores the widely admitted case-by-case approach, and confuses potential hazard and proven risk in the scientific procedure of risk assessment. Furthermore, we did not find any justification for this suspension in our extensive survey of the scientific literature regarding possible effects under natural field conditions on non-target animals. The vast majority of the 41 articles published in 2008 and 2009 indicate no impact on these organisms and only two articles indicate a minor effect, which is either inconsistent during the planting season or represents an indirect effect. Publications from 1996 to 2008 (376 publications) and recent meta-analyses do not allow to conclude on consistent effects either. The lower abundance of some insects concerns mainly specialized enemies of the target pest (an expected consequence of its control by Bt maize). On the contrary, Bt maize have generally a lower impact than insecticide treatment. The present review demonstrates that the available meta knowledge on Cry1Ab expressing maize was ignored by the German government which instead used selected individual studies. PMID- 19548101 TI - Silencing potential of viral derived RNAi constructs in Tomato leaf curl virus AC4 gene suppression in tomato. AB - We investigated viral gene suppression in an infected tomato, by transforming it with RNA inhibition (RNAi) constructs derived from same viral gene. To develop RNAi constructs, conserved sequences ranging from 21 to 200 nt of the viral target AC4 gene of various viruses causing the tomato leaf curl disease were chosen. The double-stranded (ds)RNA producing constructs carry the sense and antisense portions of these sequences and are separated by different introns behind a constitutive promoter. We compared the levels of suppression of the viral target gene by transforming four different RNAi constructs with varied arm length of dsRNA. Gene silencing levels of the viral target gene were found to be directly proportional to the arm length of the dsRNA. We observed that dsRNA derived from longer arm-length constructs generating a pool of siRNAs that were more effective in targeting gene silencing. After transformation, one of the RNAi construct having a 21 nt arm-length produced aberrant phenotypes. These phenotypic anomalies may be due to unintended ('off-target') host transcript silencing. The unintended host transcript silencing showed modest reversion in the presence of the viral target gene. The findings presented here suggest that the arm length of dsRNA capable of producing a pool of diced siRNAs is more efficient in gene silencing, the effect of off-targeting siRNA is minimized in a pool, and off-targeting silencing can be minimized in the presence of target gene. PMID- 19548102 TI - A report on the prevalence of gastrointestinal parasites in yaks (Bos poephagus) in the cold desert area of North Sikkim, India. AB - Faecal samples were collected from 348 yaks(Bos poephagus) in and around Gurudogmer Plateau, a cold desert area in North Sikkim, India. The prevalence of gastrointestinal parasitic infection was recorded at 10.05% in both the organized and traditionally managed farms of yak. The pattern of infection was either single (2.58%) or mixed (7.47%) with a faecal egg count range of 100-200 eggs per gram of faeces in positive animals. Among the helminths, Haemonchus spp. infection was predominant (6.89%) followed by Nematodirus spp. (1.72%), Cooperia spp. (1.43%) and Dicrocoelium spp. (0.29%). PMID- 19548104 TI - Effects of feeding different levels of cooked and sun dried fish offal on carcass traits of growing Rhode Island Red chicks. AB - Fourteen days old Rhode Island Red chicks were used to evaluate effects of processed fish offal (fishmeal) on DM intake (DMI), body weight gain (BWG), feed conversion ratio (FCR = DMI/BWG) and carcass traits. Ten chicks were assigned to each of 3 replicates of 6 diets (T(1), T(2), T(3), T(4), T(5), T(6), containing 0, 3.32, 6.64, 9.96, 13.28 and 16.6% fishmeal, respectively, having 18.44-19.82% CP). After 77 days of ad-lib feeding, 3 male and 3 female chicks per treatment were fasted overnight, killed and body parts weighed. T(1) had lowest but T(6) highest (p < or = 0.001) daily intakes of 68.5 and 77.0 g DM head(-1), 13.3 and 14.8 g CP head(-1) and 231 and 243 kcal ME head(-1), respectively. BWG (10.7 g head(-1) d(-1)), FCR (6.79), weights of carcass (569 g), breast (160 g) and total edible (676 g) and dressing % (66%) of T(1) (0% fishmeal) was smaller (p < or = 0.001) than those of fishmeal groups {12.8 to 13.5 g head(-1) d(-1); 5.83 to 6.35 g; 671 to 729 g; 196 to 219 g; 807 to 876 g and 67 to 68%, respectively}. Best results of BWG, FCR and carcass traits were obtained at 9.96% fishmeal inclusion level; however acceptable results could be obtained up to 12.5%. PMID- 19548103 TI - Current situation of Peste des petits ruminants (PPR) in the Sudan. AB - The current situation of PPR in Sudan was investigated. A total of 61 tissue samples were collected from various PPR suspected outbreaks in sheep in Sudan during 2008. Collected tissue samples were tested for PPR antigen using IcELISA, PPR antigen was detected in 26 out of 61 samples (42.6%). Highest antigen detection rate was in specimens collected from western Sudan. A total of 1198 serum samples were collected from sheep (n = 500), camels (n = 392), and goats (n = 306) from different areas in Sudan (Khartoum, Gezira, Tambool, River Nile, Kordofan, White Nile, Blue Nile, Gedarif, Kassala, Halfa ElGadida, Port Sudan). Collected sera were examined for PPR antibodies using cELISA, a total of 336 (67.2%) sheep, 170 (55.6%) goat and 1 (0.3%) camel samples were found to be positive. PMID- 19548105 TI - Study on caprine and ovine dermatophilosis in Wollo, Northeast Ethiopia. AB - A study on dermatophilosis in sheep (n = 1432) and goats (n = 1128) was conducted in Northeast Ethiopia. Out of 2560 examined animals, 55 (2.14%) had clinical dermatophilosis. The respective prevalence in sheep and goats were 1.5% and 2.9%. There was no significant difference (p > 0.05) in prevalence between sheep and goats and different sexes in both species. In goats, the prevalence in young (8.7%) was significantly (p < 0.05) higher than in adults (2.3%). Clinical disease was associated with orf (45% in sheep and 12% in goats), pox (22% in sheep and 18% in goats) and ticks in goats (36%, 12/33). Other risk factors associated with transmission and spread of the disease were discussed. Vaccination against concurrent infections, improved management schemes to alleviate the impact of risk factors and early antibiotic treatment against clinical disease are recommended. PMID- 19548106 TI - Gastrointestinal parasites of working donkeys of Ethiopia. AB - The general prevalence and population composition of gastrointestinal and pulmonary helminths of working donkeys were studied. For the purpose 2935 working donkeys were coprologically examined for nematode and cestode, and 215 donkeys for trematode infections. Seven donkeys that died due to various health problems or were euthanased on a welfare ground were necropsied and the parasites were recovered and identified to the species level. The study was conducted during the periods 1996-1999.Coprological examination revealed 99% strongyle, 80% Fasciola, 51% Parascaris, 30% Gastrodiscus, 11% Strongyloides westeri, 8% cestodes and 2% Oxyuris equi infection prevalence. Over 55% of donkeys had more than 1000 eggs per gram of faeces (epg). Forty two different species of parasites consisting of 33 nematodes, 3 trematodes, 3 cestodes and 3 arthropod larvae were identified from postmortem examined donkeys. Among the nematodes 17 species of Cyathostominae and 7 species of Strongylinae were identified. Other parasites identified include, Habronema muscae, Draschia megastoma, Trichostrongylus axei, Strongyloides westeri, Anoplocephala perfoliata, Anoplocephala magna, Anoplocephaloides (Paranoplocephala) mamillana, Parascaris equorum, Fasciola hepatica, Fasciola gigantica, Gastrodiscus aegyptiacus, Dictyocaulus arnfieldi, Oxyuris equi, Probstmayria vivipara, Gasterophilus intestinalis, Gasterophilus nasalis, Rhinoestrus uzbekistanicus and Setaria equina. This study revealed that working donkeys in Ethiopia are infected with a range of helminths and arthropod larvae, which are representatives of the important pathogenic parasites found in equids worldwide. PMID- 19548107 TI - Metabolic abnormalities in patients with nephrolithiasis: comparison of first episode with recurrent cases in Southern Iran. AB - BACKGROUND: Metabolic disorders are one of the etiologic factors in renal stone formation. The aim of present study was to evaluate prevalence of metabolic disorders in patients with renal stone. METHODS: From 572 patients referring to our urologic clinics, 376 patients participated in the study. Patients were divided to first time stone former (group A) and recurrent renal stone (group B). Twenty-four hour urine specimens were obtained for urinary calcium, oxalate, magnesium, citrate, uric acid, phosphor and 24-h urine volume. Venous blood samples were obtained evaluating serum phosphate, uric acid, and calcium. The prevalence of each metabolic disorder was detected, and two groups were compared regarding metabolic disorders. RESULTS: The prevalence of renal stone in male patients was 63.04% versus 36.96% in female patients (P < 0.05). The most common abnormality observed in patients was low 24-h urine volume (58.24%) followed by hypercalciuria (17.18%) and hyperuricosuria (15.15%). Hyperuricemia was found in 6 first time stone former patients and 14 patients with recurrent renal stone (P = 0.04). There was no statistically significant difference between group A and B in other metabolic abnormalities (P > 0.05). CONCLUSION: Low 24-h urine volume was the most common abnormalities in patients with nephrolithiasis in our region. Metabolic evaluation must be performed in all patients with renal stone even those with first time stone formation. PMID- 19548108 TI - Antibacterial therapy improves the effectiveness of prostate cancer detection using prostate-specific antigen in patients with asymptomatic prostatitis. AB - OBJECTIVES: To improve prostate cancer (PC) detection accuracy among patients with a prostate-specific antigen (PSA) above 4.0 ng/ml and asymptomatic prostate inflammation. MATERIALS AND METHODS: A total of 136 patients with PSA ranging from 4.0 to 50.0 ng/ml with asymptomatic prostatitis were included in the study cohort. All these patients underwent antibacterial therapy for 2 weeks followed by repeat PSA determination and transrectal ultrasound-guided needle prostate biopsy. The PSA, PSAD (PSA density), and f/t PSA (free/total PSA) before and after antibacterial therapy were compared using t-test. The receiver-operating characteristic (ROC) technique was used to evaluate the effectiveness of PSA, PSAD, f/t PSA, and their changes after antibacterial therapy (DeltaPSA, DeltaPSAD, and Deltaf/t PSA) on PC detection. RESULTS: Among the 136 patients, 33 had PC and the other 103 histologically confirmed benign prostatic disease. After antibacterial therapy for 2 weeks, the PSA (mean +/- standard deviation) decreased from 14.0 +/- 7.8 ng/ml to 10.4 +/- 7.7 ng/ml (P < 0.01). The DeltaPSA, DeltaPSAD, and Deltaf/tPSA were -3.60 +/- 4.3 ng/ml, -0.1 +/- 0.1 ng/ml/ml, and 0.1 +/- 0.1 respectively. The areas under ROC curve were 0.29 for PSA, 0.64 for PSAD, and 0.50 for f/t PSA. The areas under ROC curve were 0.91 for DeltaPSA, 0.96 for DeltaPSAD, and 0.98 for Deltaf/t PSA. These values were increased significantly when compared with those for the PSA-related parameters before antibacterial therapy (P value, DeltaPSA, DeltaPSAD, and Deltaf/t PSA were <0.01). CONCLUSIONS: Using the changes in PSA-related parameters after antibacterial therapy DeltaPSA, DeltaPSAD, and Deltaf/t PSA improve the PC detection rate and decrease unnecessary prostate biopsies in patients with asymptomatic prostatitis. PMID- 19548109 TI - The comparative study of serum iron, copper, and zinc levels between bladder cancer patients and a control group. AB - OBJECTIVE: A relatively wide range of trace elements are known to play important roles in biological processes, including the oxidative processes. Oxidative processes are one of the mechanisms involved in both incidence and recurrence of bladder cancer. In the present study, the concentration of iron (Fe), copper (Cu) and zinc (Zn) were determined in the serum of patients with bladder cancer in comparison to healthy subjects. MATERIALS AND METHODS: This cross-sectional study was conducted on 51 patients with bladder cancer and 58 healthy volunteers after age, sex, and smoking habits were matched. After overnight fasting, samples were collected. The concentrations of Fe, Cu, and Zn were measured by flame atomic absorption spectroscopy and comparisons were made using Student's t-test. RESULTS: There was a significant increase in mean Cu and Cu/Zn serum level in bladder cancer patients compared to the control group (P < 0.001). In contrast, the serum zinc level in patients having bladder cancer was significantly lower than in the control group (P < 0.05). Moreover, the serum iron level was significantly lower in the patients than the control group (P < 0.001). CONCLUSIONS: In the present study, a relationship was seen between the level of trace elements and the occurrence of bladder cancer, suggesting that an increase in the serum level of Cu and a decrease in the levels of Zn and Fe might be important causes of bladder cancer occurrence; however, defining such a cause-and effect relationship needs several prospective studies to be done, which seems necessary with regard to the high prevalence of this cancer. PMID- 19548110 TI - Exploring photosynthesis by electron tomography. PMID- 19548112 TI - In vitro interactions between antifungals and methotrexate against Aspergillus spp. AB - Methotrexate has been widely used in the treatment of osteosarcoma, intracranial lymphomas and leukemia. However, patients are also at high risk of opportunist pathogens such as Aspergillus spp. infection for their deeply depressed immunity. The optimal choice of antifungal agents during the infection of Aspergillus for these patients is necessary to be explored. In this study, we investigated the interactions between antifungals and methotrexate against Aspergillus in vitro. A total of 23 clinical isolates of Aspergillus spp. were studied. Microdilution checkerboard technique was performed to evaluate the interaction of methotrexate with voriconazole, itraconazole, terbinafine and amphotericin B. The highest rate of synergy was obtained for the combination of terbinafine and methotrexate, which exhibited synergy against 60.9% (14/23) of strains. No interaction was detected for the combinations of methotrexate plus itraconazole or amphotericin B against 95.7% (22/23) or 100% of strains, respectively. Although voriconazole exhibited indifferent against 87% (20/23) of strains when combined with methotrexate, antagonism effect was found against 13% (3/23) of strains. The positive interactions of terbinafine and methotrexate were also certified by disk diffusion assay. In addition, we observed the morphological changes for the interaction of methotrexate with terbinafine against Aspergillus. Further inhibition and distortion of growth were found after the combination of terbinafine and methotrexate compared with the drugs treated alone. Clinical studies are warranted to further elucidate optimal treatments for the immunocompromised patients with Aspergillus infection. PMID- 19548111 TI - Delayed fluorescence in photosynthesis. AB - Photosynthesis is a very efficient photochemical process. Nevertheless, plants emit some of the absorbed energy as light quanta. This luminescence is emitted, predominantly, by excited chlorophyll a molecules in the light-harvesting antenna, associated with Photosystem II (PS II) reaction centers. The emission that occurs before the utilization of the excitation energy in the primary photochemical reaction is called prompt fluorescence. Light emission can also be observed from repopulated excited chlorophylls as a result of recombination of the charge pairs. In this case, some time-dependent redox reactions occur before the excitation of the chlorophyll. This delays the light emission and provides the name for this phenomenon-delayed fluorescence (DF), or delayed light emission (DLE). The DF intensity is a decreasing polyphasic function of the time after illumination, which reflects the kinetics of electron transport reactions both on the (electron) donor and the (electron) acceptor sides of PS II. Two main experimental approaches are used for DF measurements: (a) recording of the DF decay in the dark after a single turnover flash or after continuous light excitation and (b) recording of the DF intensity during light adaptation of the photosynthesizing samples (induction curves), following a period of darkness. In this paper we review historical data on DF research and recent advances in the understanding of the relation between the delayed fluorescence and specific reactions in PS II. An experimental method for simultaneous recording of the induction transients of prompt and delayed chlorophyll fluorescence and decay curves of DF in the millisecond time domain is discussed. PMID- 19548113 TI - Analysis of two sequence variants in peroxisome proliferator activated receptor gamma gene in Tajik population at high altitudes and Han population at low altitudes in China. AB - The Tajik people in China have resided at high altitude for thousands of years. We analyzed the Pro12Ala (C > G) polymorphism in exon B and the 161C > T polymorphism in exon 6 of peroxisome proliferator activated receptor gamma gene (PPARG) in Chinese Tajik population living at high altitude and Chinese Han population living at low attitude. Significant higher frequencies of the CG and GG genotypes and G allele of the Pro12Ala (C > G) polymorphism were observed in the Tajik population than that in the Han population (P < 0.0001), which suggested the G allele was associated with high-altitude adaptation in the dominate model. The significant differences were remained in both of the male and female groups after stratified by gender, and the differences were more pronounced in men (G versus C, OR = 7.700) than in women (OR = 5.056). No significant difference was observed for the 161C > T polymorphism in the two populations. The frequencies of haplotypes GT (P < 0.0001) and GC (P < 0.05) were significantly higher, while the frequency of CT (P < 0.0001) was significantly lower in the Tajik population than that in the Han population. Our results suggest that PPARG is a candidate gene for high-altitude adaptation in the Chinese Tajik population. PMID- 19548114 TI - Clinical reasoning: new challenges. AB - This article is an introduction to a special issue of Theoretical Medicine and Bioethics on clinical reasoning. Clinical reasoning encompasses the gamut of thinking about clinical medical practice--the evaluation and management of patients' medical problems. Theories of clinical reasoning may be normative or descriptive; that is, they may offer recommendations on how clinicians ought to think or they may simply attempt to describe how clinicians actually do think. This article briefly surveys these approaches in order to show the complexity of clinical reasoning and the inadequacy of any one theory for capturing the full richness of clinical reasoning. The authors of this issue offer both normative and descriptive elements in their accounts. Topics discussed include the importance for clinical reasoning of tacit knowing, risk assessment, narrative and hermeneutics, wisdom, and virtue epistemology. PMID- 19548115 TI - The epistemically virtuous clinician. AB - Today, modern Western medicine is facing a quality-of-care crisis that is undermining the patient-physician relationship. In this paper, a notion of the epistemically virtuous clinician is proposed in terms of both the reliabilist and responsibilist versions of virtue epistemology, in order to help address this crisis. To that end, a clinical case study from the literature is first reconstructed. The reliabilist intellectual virtues, including the perceptual and conceptual virtues, are then discussed and applied to the case study. Next, a similar method is employed to examine the responsibilist intellectual virtues, including curiosity, courage, honesty, and humility, and to apply them to the case study. To round out the discussion, the love of knowledge and both theoretical and practical wisdom are explored and applied to the case study. The paper concludes with a brief discussion of how the notion of an epistemically virtuous clinician addresses the quality-of-care crisis, in terms of the connection between ethical and intellectual virtues, and of the notion's implications for medical education. PMID- 19548116 TI - Clinical intuition versus statistics: different modes of tacit knowledge in clinical epidemiology and evidence-based medicine. AB - Despite its phenomenal success since its inception in the early nineteen nineties, the evidence-based medicine movement has not succeeded in shaking off an epistemological critique derived from the experiential or tacit dimensions of clinical reasoning about particular individuals. This critique claims that the evidence-based medicine model does not take account of tacit knowing as developed by the philosopher Michael Polanyi. However, the epistemology of evidence-based medicine is premised on the elimination of the tacit dimension from clinical judgment. This is demonstrated through analyzing the dichotomy between clinical and statistical intuition in evidence-based medicine's epistemology of clinical reasoning. I argue that clinical epidemiology presents a more nuanced epistemological model for the application of statistical epidemiology to the clinical context. Polanyi's theory of tacit knowing is compatible with the model of clinical reasoning associated with clinical epidemiology, but not evidence based medicine. PMID- 19548117 TI - Quality of life from the perspective of adolescents with cerebral palsy: "I just think I'm a normal kid, I just happen to have a disability". AB - PURPOSE: The aim of this qualitative study was to obtain the perspectives of adolescents with cerebral palsy (CP) on factors that positively or negatively influence their quality of life (QoL). METHODS: Using constructivist grounded theory, adolescents (12-16 years) were interviewed to obtain their accounts of intrinsic and extrinsic attributes that made a difference in their satisfaction with life. A purposive and theoretical sample of 12 adolescents was obtained. Narratives were coded into categories and themes, which were initially broadly defined as intrinsic (child-related) and extrinsic (environmental) factors, and then were further differentiated as analysis progressed. RESULTS: Participants reflected on their quality of life as an interaction between intrinsic and extrinsic factors. The predominant theme that emerged involved a relationship between personal interests and preferences (intrinsic) and opportunities to participate in age-appropriate activities and leisure activities (extrinsic). Opportunities to participate in activities that the adolescents value affect their perceived quality of life and this perception was consistent regardless of the objective level of motor impairment. CONCLUSION: Health care providers should take into consideration what adolescents with CP value the most. The identification of attributes that can be modified such as accessibility to and competence in, and preference for different activities may help professionals to advocate for greater participation in these activities and tailor individual therapeutic strategies accordingly, increasing the likelihood of enhancing the quality of life of this population. PMID- 19548118 TI - Building PROMIS item banks: librarians as co-investigators. AB - PURPOSE: There is growing interest in the use of item response theory (IRT) for creation of measures of health-related quality of life (HRQOL). A first step in IRT modeling is development of item banks. Our aim is to describe the value of including librarians and to describe processes used by librarians, in the creation of such banks. METHOD: Working collaboratively with PROMIS researchers at the University of Pittsburgh, a team of librarians designed and implemented comprehensive literature searches in a selected set of information resources, for the purpose of identifying existing measures of patient-reported emotional distress. RESULTS: A step-by-step search protocol developed by librarians produced a set of 525 keywords and controlled vocabulary terms for use in search statements in 3 bibliographic databases. These searches produced 6,169 literature citations, allowing investigators to add 444 measurement scales to their item banks. CONCLUSION: Inclusion of librarians on the Pittsburgh PROMIS research team allowed investigators to create large initial item banks, increasing the likelihood that the banks would attain high measurement precision during subsequent psychometric analyses. In addition, a comprehensive literature search protocol was developed that can now serve as a guide for other investigators in the creation of IRT item banks. PMID- 19548119 TI - Antioxidant activity of sulfur and selenium: a review of reactive oxygen species scavenging, glutathione peroxidase, and metal-binding antioxidant mechanisms. AB - It is well known that oxidation caused by reactive oxygen species (ROS) is a major cause of cellular damage and death and has been implicated in cancer, neurodegenerative, and cardiovascular diseases. Small-molecule antioxidants containing sulfur and selenium can ameliorate oxidative damage, and cells employ multiple antioxidant mechanisms to prevent this cellular damage. However, current research has focused mainly on clinical, epidemiological, and in vivo studies with little emphasis on the antioxidant mechanisms responsible for observed sulfur and selenium antioxidant activities. In addition, the antioxidant properties of sulfur compounds are commonly compared to selenium antioxidant properties; however, sulfur and selenium antioxidant activities can be quite distinct, with each utilizing different antioxidant mechanisms to prevent oxidative cellular damage. In the present review, we discuss the antioxidant activities of sulfur and selenium compounds, focusing on several antioxidant mechanisms, including ROS scavenging, glutathione peroxidase, and metal-binding antioxidant mechanisms. Findings of several recent clinical, epidemiological, and in vivo studies highlight the need for future studies that specifically focus on the chemical mechanisms of sulfur and selenium antioxidant behavior. PMID- 19548120 TI - Acute lung injury in patients with severe brain injury: a double hit model. AB - The presence of pulmonary dysfunction after brain injury is well recognized. Acute lung injury (ALI) occurs in 20% of patients with isolated brain injury and is associated with a poor outcome. The "blast injury" theory, which proposes combined "hydrostatic" and "high permeability" mechanisms for the formation of neurogenic pulmonary edema, has been challenged recently by the observation that a systemic inflammatory response may play an integral role in the development of pulmonary dysfunction associated with brain injury. As a result of the primary cerebral injury, a systemic inflammatory reaction occurs, which induces an alteration in blood-brain barrier permeability and infiltration of activated neutrophils into the lung. This preclinical injury makes the lungs more susceptible to the mechanical stress of an injurious ventilatory strategy. Tight CO2 control is a therapeutic priority in patients with acute brain injury, but the use of high tidal volume ventilation may contribute to the development of ALI. Establishment of a therapeutic regimen that allows the combination of protective ventilation with the prevention of hypercapnia is, therefore, required. Moreover, in patients with brain injury, hypoxemia represents a secondary insult associated with a poor outcome. Optimal oxygenation may be achieved by using an adequate FiO2 and by application of positive end-expiratory pressure (PEEP). PEEP may, however, affect the cerebral circulation by hemodynamic and CO2-mediated mechanisms and the effects of PEEP on cerebral hemodynamics should be monitored in these patients and used to titrate its application. PMID- 19548122 TI - Can lupus flares be associated with tuberculosis infection? AB - Systemic lupus erythematosus (SLE) is an autoimmune disease that frequently requires treatment with high doses of corticosteroids and immunosuppressive drugs. Primary defects in the innate immunity also contribute to an increased susceptibility to infections. Patients with SLE are at an increased risk for infections with several pathogens, among them Mycobacterium tuberculosis, which is a significant cause of morbidity and mortality, especially in endemic regions. TB infection requires awareness for several reasons: first, TB infection thrives under conditions of immunosuppression, may it be secondary to the disease itself or its treatment. Second, shared antigens by mycobacteria and autoantigens have been described, which may be targets for autoantibodies. We present four Brazilian patients, in whom a diagnosis of tuberculosis was determined during or following persistent flares of their disease. The association of SLE and TB is discussed, as well as different aspects of the tuberculosis infection in this selected population, and its possible role in the course of SLE. PMID- 19548121 TI - Antifibrosis: to reverse the irreversible. AB - Fibrosis is a pathological process that includes scar formation and overproduction of extracellular matrix by the connective tissue as a response to tissue damage. The fibrotic process involves multiple organs and results in progressive life-threatening diseases. Today, we know more about the molecular mechanism that leads to fibrosis involving different type of cells, cytokines, chemokines, and tissue enzymes. Fibrosis was considered an irreversible process, at least clinically, and is still usually treated by anti-inflammatory and immunosuppressive agents. No proven antifibrotic therapy has shown efficacy in ameliorating the clinical course of fibrotic diseases, but our current understanding led to the development of different drugs with promising results, like: mycophenolate mofetil, interferon, relaxin, and intravenous immunoglobulin. This review will provide a glance to this heavily investigated subject. PMID- 19548123 TI - Leukocyte immunoglobulin-like receptors as new players in autoimmunity. AB - Leukocyte immunoglobulin-like receptors (LILR) are a family of at least 13 receptors mainly expressed on lymphoid and myelomonocytic cells. They are involved in the activation of the immune system. Inhibitory LILR (termed LILRB) signal through immunoreceptor tyrosine-based inhibitory motives in the cytoplasmic domain, whereas LILRA with short cytoplasmic domains are stimulatory receptors. Polymorphisms and deletions of leukocyte immunoglobulin-like receptors have been shown to be associated with autoimmune disorders, and some of the receptors are involved in the generation of regulatory T cells. Therefore, leukocyte immunoglobulin-like receptors may be central in the pathogenesis of autoimmunity. The data linking these receptors to autoimmune diseases is reviewed here. PMID- 19548124 TI - Efficacy and safety of anti-tumour necrosis factor in elderly patients with rheumatoid arthritis: an observational study. AB - This study aims to compare the efficacy and safety of anti-TNF agents in elderly (aged >or=65 years) and younger patients (aged 18-65 years) with active RA. The study involved 1,114 RA patients treated with anti-TNF drugs and followed-up for >6 months by LORHEN group, who were divided into two cohorts on the basis of their age (311 aged >or=65 and 803 aged <65 years) in order to evaluate 3-year outcomes and treatment discontinuations. Drug effectiveness was assessed by disease activity (DAS28 and EULAR response), functional status (HAQ) and serological parameters (ESR) at baseline and during anti-TNFalpha therapy; safety was evaluated on the basis of drug discontinuation rates. At baseline, the elderly patients showed greater disease activity (DAS28, ESR) and loss of joint function (HAQ, functional class; p < 0.05). During therapy, clinical and laboratory parameters (DAS28, ESR) improved in both groups without any statistically significant difference between them, whereas the difference in HAQ remained after 36 months of treatment (p < 0.05). Anti-TNFalpha therapy was discontinued by 123 of the elderly (42%) and 282 of the younger patients (36.6%) because of loss of efficacy (17.4% vs. 16.7%), severe adverse events (21.8% vs. 16.9%) or other reasons (2.7% vs. 3%). The number of adverse events was significantly higher in the elderly patients (p < 0.05). Anti-TNFalpha treatment reduced disease activity and led to functional improvement in both groups, although the baseline difference in HAQ remained statistically significant at the end of the follow-up. The elderly patients experienced more infective events. PMID- 19548125 TI - Verification of a prolonged untreated natural history of breast cancer by the multigene assay. AB - Individualization of therapy for breast cancer patients has progressed significantly over the last 5 years. A 54-year-old female went over 2 years after her diagnosis of breast cancer with no treatment. The pathologic size, however, indicated that the tumor may not have progressed from diagnosis to operation. Due to the apparent lack of progression over 2 years without treatment, a multigene assay was ordered. The recurrence score was 15, indicating a less than 10% risk of distant recurrence at 10 years. The recurrence score also falls into the "low risk" category. The prolonged natural history of this breast cancer patient suggested an indolent cancer. The "low risk" recurrence score confirmed this history and indicated that adjuvant chemotherapy is probably not beneficial to this patient. PMID- 19548126 TI - Bone marrow hyaluronan and reticulin in patients with malignant disorders. AB - Myelofibrosis is commonly seen in patients with chronic myeloproliferative diseases and sometimes in myelodysplastic syndrome, acute leukaemia and lymphoproliferative diseases. The fibrotic process is evaluated by grading the amount of collagen deposited in the bone marrow interstitium. The established method to evaluate bone marrow fibrosis is staining for reticulin to visualise the collagen fibres. However, the extra cellular matrix does not only contain collagens but also other components, e.g. glycosaminoglycans of which hyaluronan is the most abundant. Hyaluronan is important for structural and cellular functions. Earlier studies have shown that there is a positive correlation between hyaluronan and reticulin staining in healthy volunteers and in patients with de novo acute myeloid leukaemia. In this study bone marrow biopsies from 43 patients with a malignant disease involving the bone marrow were compared with 18 patients with a malignant disease not involving the bone marrow. The intensity of hyaluronan grading was significantly higher in the patients with disease involving the bone marrow compared to the healthy controls but not compared to the patients without disease involving the bone marrow. The staining intensity of reticulin in the bone marrow was significantly higher in the patients with disease involving the bone marrow, compared to those without disease involving the bone marrow and to the controls. In all patients and the controls there was a correlation between hyaluronan and reticulin. PMID- 19548128 TI - Xenia Forsselliana 2008. PMID- 19548127 TI - c-erbB-2 and the "triple-state" in early breast carcinomas. AB - Although c-erbB-2 expression is, in general terms, an ominous prognostic indicator in breast carcinomas, there are suggestions that lack of this oncogene, when combined with analogous lack of estrogen (ER negative) and progesterone receptors (PgR negative)-"triple-negative phenotype", is linked with an equally poor prognosis. We investigated this hypothesis in a series of early ductal breast carcinomas. A total of 116 specimens with early breast cancer, defined as tumors of < or =2 cm in size and clinically negative axilla, were studied immunohistochemically for ER, PgR, and c-erbB-2 expression. The median follow-up was 131 months (range 62-245 months). ER positive tumors had a favorable clinical course, compared to ER negative neoplasms, but only for the first 10 years of follow-up (P = 0.04). Prognosis was poorer for the PgR negative cases, relative to PgR positive tumors (P = 0.005), but this stood true for the entire investigation period. Triple-negative breast carcinomas had a poor prognosis, while triple-positive tumors had a favorable outcome. However, if triple-positive and triple-negative cases were excluded from the original sample, the remaining c erbB-2 positive cases were connected with poor prognosis, relative to the remaining c-erbB-2 negative tumors. c-erbB-2 oncogene has a complex biological role in early breast carcinomas for its expression characterizes subgroups of patients with both favorable (triple-positive phenotype) and unfavorable prognosis (c-erb-B2 positive cases after excluding triple-positive and triple negative tumors)-a phenomenon presumably due to activation of different biological pathways. Elucidation of these pathways may determine subgroups of patients with tumors requiring different targeted agents. PMID- 19548129 TI - Acute hyperglycemia and spontaneous reperfusion in acute myocardial infarction. AB - BACKGROUND: Spontaneous reperfusion (SR) may occur in patients with ST elevation myocardial infarction (STEMI) prior to reperfusion therapy. Hyperglycemia is common on admission in patients with STEMI and is associated with a worse prognosis. Mechanisms remain unclear but may include impairment of coronary flow. The objective of this study was to examine whether acute hyperglycemia influenced the occurrence of SR in patients with STEMI. METHODS: All patients presenting to our institution with acute STEMI with measurement of glucose levels on presentation were eligible. SR was defined as a combination of significant relief of chest pain associated with an at least 70% resolution of ST segment elevation on follow-up ECG. RESULTS: 465 patients were studied of whom 77 patients met criteria for SR. Average glucose levels were not significantly different between the SR and non-SR groups (10.0+/-5.6 mmol/l versus 10.1+/-5.3; P=NS). When patients were divided into normoglycemic and hyperglycemic groups, there was no significant difference in the percentages of such patients in the SR and non-SR groups. (52% versus 54%; P=NS). CONCLUSIONS: Acute hyperglycemia on admission does not predict the occurrence of SR in a general population of patients with acute MI. PMID- 19548130 TI - Temporary (short-term) percutaneous left ventricular assist device (Tandem Heart) in a patient with STEMI, multivessel coronary artery disease, cardiogenic shock and severe peripheral artery disease. AB - Cardiogenic shock remains the leading cause of mortality among patients admitted for acute myocardial infarction (AMI). Percutaneous left ventricular assist devices (pVAD) represent one possible means of supporting these critically ill patients during interventional procedures. Tandem Heart is one such pVAD, which uses transseptal cannulation and a paracorporal centrifugal pump to maintain active circulation to the descending aorta. Despite its overall effectiveness in clinical use, the use of the Tandem Heart has several potential limitations, including a contraindication for patients with peripheral artery disease. This case report describes the successful use of the Tandem Heart in one such patient requiring a high-risk coronary intervention procedure with otherwise prohibitive peripheral arterial disease, by employing peripheral artery angioplasty. PMID- 19548131 TI - In vitro comparison of dodecafluoropentane (DDFP), perfluorodecalin (PFD), and perfluoroctylbromide (PFOB) in the facilitation of oxygen exchange. AB - The purpose is to prepare 2% w/v emulsions of dodecafluoropentane, perfluorodecalin, and perfluoroctylbromide and compare them for their ability to absorb oxygen. The oxygen uptake capability and volume expansion of each emulsion and the blank vehicle were evaluated in water at 21 degrees C and 37 degrees C. The average particle size of the dodecafluoropentane emulsion is < 400 nm stored at room temperature for 6 months. In comparison to water treated with either the blank vehicle, the perfluorodecalin emulsion, or the perfluoroctylbromide emulsion, the dodecafluoropentane emulsion absorbs 3 times more oxygen at 21 degrees C and 7 times more oxygen at 37 degrees C. Furthermore, a significantly higher in vitro expansion (5 times) is observed with the dodecafluoropentane emulsion at 37 degrees C. As such, DDFP has been hypothesized to be a better oxygen carrier and delivery agent in vivo. This may be applicable to a variety of hypoxic medical conditions where oxygen delivery might be therapeutically beneficial. PMID- 19548133 TI - Hemoglobin-based oxygen carriers combined with anticancer drugs may enhance sensitivity of radiotherapy and chemotherapy to solid tumors. AB - The aberrant vascular architecture in solid tumors is the key limiting factor known to ameliorate hypoxia and increase circulating anticancer drugs, thus resulting in resistance to radiotherapy and chemotherapy in tumor treatment. Previous experiments have reported hemoglobin-based oxygen carriers (HBOCs) that are effective to improve tumor oxygenation, thereby serving as potential agents target-oriented to the hypoxic tumor. Herein, we draw the hypothesis that HBOCs combined with an anticancer drug may increase oxygen bioavailability and anticancer drug retention in solid tumors and in turn contribute to enhanced sensitivity of radiotherapy and chemotherapy. This novel drug will bring a new breakthrough in the field of the development of anticancer drugs and reveal the alternative clinical use of HBOCs in tumor treatment. PMID- 19548134 TI - A special issue devoted to hematoxylin, hematein, and hemalum. PMID- 19548132 TI - Soluble angiopoietin receptor Tie-2 in patients with acute myocardial infarction and its detection by optical protein-chip. AB - The Tie-2 receptor has been shown to play a role in angiogenesis in atherosclerosis. The conventional method assaying the level of soluble Tie-2 (sTie-2) was ELISA. However, this method has some disadvantages. The aims of this research are to establish a more simple detection method, the optical protein chip based on imaging ellipsomtry (OPC-IE) applying to Tie-2 assay. The sTie-2 biosensor surface on silicon wafer was prepared first, and then serum levels of sTie-2 in 38 patients with AMI were measured on admission (day 1), day 2, day 3 and day 7 after onset of chest pain and 41 healthy controls by ELISA and OPC-IE in parallel. Median level of sTie-2 increased significantly in the AMI patients when compared with the controls. Statistics showed there was a significant correlation in sTie-2 results between the two methods (r=0.923, P<0.01). The result of this study showed that the level of sTie-2 increased in AMI, and OPC-IE assay was a fast, reliable, and convenient technique to measure sTie-2 in serum. PMID- 19548138 TI - CD147 and VEGF expression in advanced renal cell carcinoma and their prognostic value. AB - AIM: To investigate the clinicopathologic characteristics of extracellular matrix (ECM) metalloproteinase inducer (CD147) and vascular endothelial growth factor (VEGF) expression in advanced renal cell carcinoma (RCC), and to evaluate the clinical significance of these two markers in the prognosis of advanced RCC. METHODS: CD147 and VEGF expression in paraffin-embedded specimens gathered from 53 patients with advanced RCC and 12 healthy controls were detected by the method of immunohistochemistry. The Spearman correlation was calculated between the expression levels of CD147 and VEGF in advanced RCC tissues. The association of CD147 and VEGF expression with the clinicopathologic features and prognosis of advanced RCC was subsequently assessed. RESULTS: CD147 and VEGF were positively expressed in 47/53 (88.7%) and 45/53 (84.9%) of patients with advanced RCC, respectively. Positive expression of CD147 (p= 0.02) and VEGF (p< 0.01) was significantly correlated with TNM stage of advanced RCC. A significant correlation was found between the expression of CD147 and VEGF in advanced RCC (r= 0.629, p= 0.04). Additionally, tumor CD147 and tumor VEGF expressions were significantly associated with the prognosis of advanced RCC patients. The survival rate of the patients with CD147-/VEGF- expression was the lowest (p< 0.01), and conjoined expressions of CD147-/VEGF- and CD147+/VEGF+ were independent prognostic indicators of advanced RCC (both p< 0.01). CONCLUSION: The expression of CD147 or VEGF may be an important feature of advanced RCC. A combined detection of CD147/VEGF coexpression may benefit us in the prediction of the prognosis of advanced RCC. PMID- 19548140 TI - Utility of gene promoter methylation in prediction of response to platinum-based chemotherapy in epithelial ovarian cancer (EOC). AB - The aim was to determine whether promoter methylation of BRCA1, MGMT, MLH1, RASSF1A, and p16 genes could predict response to platinum-based chemotherapy. Thirty-five subjects with epithelial ovarian cancer (EOC) treated by platinum based chemotherapy were recruited. Methylation-specific polymerase chain reaction was carried out and the methylation index (MI) was also derived. Response to platinum-based chemotherapy was documented clinically, radiologically, and by serial CA125 levels. Methylated BRCA1 (p = .037) and a higher MI (p = .045) were associated with primary chemosensitivity. A better outcome was predicted by a higher MI (p = .032). In EOC, BRCA1 gene promoter methylation is useful in the prediction of response to chemotherapy. PMID- 19548142 TI - Human embryonic stem cell-derived neural precursor transplants in collagen scaffolds promote recovery in injured rat spinal cord. AB - BACKGROUND AIMS: Several studies have reported functional improvement after transplantation of in vivo-derived neural progenitor cells (NPC) into injured spinal cord. However, the potential of human embryonic stem cell-derived NPC (hESC-NPC) as a tool for cell replacement of spinal cord injury (SCI) should be considered. METHODS: We report on the generation of NPC as neural-like tubes in adherent and feeder-free hESC using a defined media supplemented with growth factors, and their transplantation in collagen scaffolds in adult rats subjected to midline lateral hemisection SCI. RESULTS: hESC-NPC were highly expressed molecular features of NPC such as Nestin, Sox1 and Pax6. Furthermore, these cells exhibited the multipotential characteristic of differentiating into neurons and glials in vitro. Implantation of xenografted hESC-NPC into the spinal cord with collagen scaffold improved the recovery of hindlimb locomotor function and sensory responses in an adult rat model of SCI. Analysis of transplanted cells showed migration toward the spinal cord and both neural and glial differentiation in vivo. CONCLUSIONS: These findings show that transplantation of hESC-NPC in collagen scaffolds into an injured spinal cord may provide a new approach to SCI. PMID- 19548144 TI - Multipotent mesenchymal stem cells from amniotic fluid originate neural precursors with functional voltage-gated sodium channels. AB - BACKGROUND AIMS: Amniotic fluid (AF) contains stem cells with high proliferative and differentiative potential that might be an attractive source of multipotent stem cells. We investigated whether human AF contains mesenchymal stem cells (MSC) and evaluated their phenotypic characteristics and differentiation potential in vitro. METHODS: AF was harvested during routine pre-natal amniocentesis at 14-16 weeks of pregnancy. AF sample pellets were plated in alpha minimum essential medium (MEM) with 10% fetal bovine serum (FBS). We evaluated cellular growth, immunophenotype, stemness markers and differentiative potential during in vitro expansion. Neural progenitor maintenance medium (NPMM), a medium normally used for the growth and maintenance of neural stem cells, containing hFGF, hEGF and NSF-1, was used for neural induction. RESULTS: Twenty-seven AF samples were collected and primary cells, obtained from samples containing more than 6 mL AF, had MSC characteristics. AF MSC showed high proliferative potential, were positive for CD90, CD105, CD29, CD44, CD73 and CD166, showed Oct 4 and Nanog molecular and protein expression, and differentiated into osteoblasts, adypocytes and chondrocytes. The NPMM-cultured cells expressed neural markers and increased Na(+) channel density and channel inactivation rate, making the tetrodotoxin (TTX)-sensitive channels more kinetically similar to native neuronal voltage-gated Na(+) channels. CONCLUSIONS: These data suggest that AF is an important multipotent stem cell source with a high proliferative potential able to originate potential precursors of functional neurons. PMID- 19548145 TI - Body awareness, eating attitudes, and spiritual beliefs of women practicing yoga. AB - This research evaluated attitudes about body image and eating in women practicing postural yoga. Study 1 described scores from questionnaires on variables related to body awareness, intuitive eating, spirituality, and reasons for practicing. Scores were favorable on all measures with significant correlations (p < .01) among all main variables except between spiritual readiness and intuitive eating, and between BMI and both body awareness and spiritual readiness. Reasons for practicing did not affect scores. Study 2 evaluated interviews in a sub-sample. Qualitative data reported improvements in body satisfaction and disordered eating due in part to yoga and its associated spirituality. PMID- 19548146 TI - A demographic and symptom descriptive study of callers to the National Association of Anorexia Nervosa and Associated Disorders: linking research and advocacy. AB - Phone support hotlines are an invaluable service to many eating disorder sufferers and their loved ones, and can also provide important data for eating disorder research. In the present study, the demographic and symptom information of nearly 800 volunteer callers to a free national eating disorder support and referral hotline was analyzed. Findings indicate that eating disorder sufferers who are referenced through hotline resources tend to be older, come from more varied socioeconomic backgrounds, and have more atypical symptoms than what is generally described in existing sources. Results are discussed in light of future directions for hotline research. PMID- 19548147 TI - Personality and psychological factors as predictors of disordered eating among female collegiate athletes. AB - This article examined the relationship of perfectionism, psychological well-being (self-esteem and optimism), reasons for exercising and appearance orientation to eating disorder classification among 204 female collegiate athletes. Multivariate analyses showed that only self-esteem, exercising to improve appearance and be more attractive, and appearance orientation differentiated significantly between the symptomatic/eating disordered athletes and those who were asymptomatic. No differences existed between the two groups of athletes on perfectionism, optimism, or exercising for fitness/health. For athletes, self-esteem, appearance orientation and exercising to be attractive and improve appearance were most important for understanding their level of disordered eating. PMID- 19548148 TI - Serum albumin levels may not correlate with weight status in severe anorexia nervosa. AB - Anorexia nervosa is a difficult disease to treat effectively. Inpatient treatment in facilities with specialized expertise heightens the chance for success. Patients with the most severe degrees of anorexia nervosa are especially in need of hospitalization. Authorization from insurers can be a barrier to admitting these patients to reputable treatment facilities. Therefore, familiarity with accurate markers of disease severity is important to understand in order to effectively advocate for these patients. Albumin, a commonly used marker for nutritional status is surprisingly normal even in patients with severe anorexia nervosa. Understanding that albumin levels do not correlate with the severity of anorexia nervosa is an important lesson to understand in the process of facilitating the most effective care settings for patients with severe anorexia nervosa. PMID- 19548149 TI - Meal support therapy reduces the use of nasogastric feeding for adolescents hospitalized with anorexia nervosa. AB - The objective of this study was to assess the impact of meal support therapy on the use of nasogastric feeding for adolescent patients hospitalized with Anorexia Nervosa. Charts of 21 adolescents (mean age 15.1 +/- 1.9 years) were reviewed. The incidence of nasogastric feeding was substantially reduced after the implementation of meal support therapy (67% versus 11% [p < .02]). No differences in length of stay, change in weight, or readmission rates were observed. Meal support therapy significantly reduces the use of nasogastric feeding on inpatient units treating children and adolescents with Anorexia Nervosa. PMID- 19548150 TI - Perceived incompetence moderates the relationship between maladaptive perfectionism and disordered eating. AB - Maladaptive perfectionism and perceived incompetence are two factors associated with disordered eating. In this study, we examined whether perceived incompetence moderated the relationship between maladaptive perfectionism and disordered eating. Three hundred fifteen college women completed surveys assessing eating habits and levels of perceived incompetence and perfectionism. Results supported a moderating effect of perceived incompetence such that as levels of perceived incompetence increased, the relationship between maladaptive perfectionism and disordered eating became increasingly strong. These results imply that clinicians may want to focus efforts on helping clients learn to set healthy goals and improve their perception of competence in life domains. PMID- 19548153 TI - Betacyanins as phenol antioxidants. Chemistry and mechanistic aspects of the lipoperoxyl radical-scavenging activity in solution and liposomes. AB - Reaction kinetics of betanin and its aglycone betanidin towards peroxyl radicals generated from the azo-initiated oxidation of methyl linoleate in methanol and of a heterogeneous aqueous/soybean phosphatidylcholine liposomal system were studied by monitoring formation of linoleic acid hydroperoxides and consumption of the pigments. Betanin was a weak retarder in methanol and an effective chain breaking antioxidant in the liposomal model, indicating that kinetic solvent effects and partition in lipid bilayers may affect its activity. Betanidin behaved as a chain terminating antioxidant in both models. Kinetic parameters characterizing peroxyl radical-scavenging activity showed that betanidin was more effective than betanin, in terms of both radical-scavenging rate constant and stoichiometric factor, with effectiveness of the same order as vitamin E under comparable conditions. Products identified by spectrophotometric and HPLC techniques indicated reaction of the glucose-substituted monophenol and ortho-diphenol moieties of betanin and betanidin, respectively, and suggested mechanisms of the antioxidant activity. Either betanin or betanidin incorporated in liposomes with alpha-tocopherol had additive effects, supporting partition of the pigments in the bilayer and lipoperoxyl radical reduction. PMID- 19548154 TI - alpha-Lipoic acid modulates thiol antioxidant defenses and attenuates exercise induced oxidative stress in standardbred trotters. AB - Several micronutrient supplementation strategies are used to cope with oxidative stress, although their benefits have recently been questioned. The aim of the present study was to examine the effects of DL-alpha-lipoic acid (LA) in response to acute exercise and during recovery in horses. Six standardbred trotters were tested on the treadmill before and after 5-week LA supplementation (25 mg/kg body weight/day). According to electron paramagnetic resonance measurements, strenuous aerobic exercise increased significantly free radical formation in the gluteus medius muscle, which was prevented by LA supplementation. The activities of thioredoxin reductase and glutathione reductase in muscle were significantly increased in LA-treated horses, but neither LA nor exercise affected muscle thioredoxin activity. LA increased the concentration of total glutathione in muscle at rest and during recovery. Treatment with LA blunted the exercise induced increase in plasma oxygen radical absorbance capacity and decreased the post-exercise levels of lipid hydroperoxides in plasma and malondialdehyde in plasma and in muscle. These findings suggest that LA enhances thiol antioxidant defences and decreases exercise-induced oxidative stress in skeletal muscle. PMID- 19548155 TI - Time-related alterations of superoxide radical levels in diverse organs of bile duct-ligated rats. AB - The time-related alterations of superoxide radical measured in vivo by employing an ultrasensitive fluorescent assay in the liver, intestine, kidney and brain of rats with experimentally induced obstructive jaundice was investigated. Eighteen rats were randomly divided into Group A, rats subjected to sham operation, and Group B, rats subjected to bile duct ligation (BDL). Three rats from each group were subsequently killed at different time points post-operatively (1, 5 and 10 days). As compared to sham-operated, BDL rats showed a gradual increase with time of superoxide radical in the intestine, liver, kidney and brain: for animals sacrificed on the 1(st), 5(th) and 10(th) day the increase was 45%, 50% and 96% in the liver, 76%, 81% and 118% in the intestine, 64%, 71% and 110% in the kidney and 76%, 95% and 142% in the brain, respectively. This study provides direct evidence of an early appearance of oxidative stress in diverse organs, implying a uniform systemic response to biliary obstruction and emphasizing the need of early bile flow restoration. PMID- 19548157 TI - HIV/AIDS, STIs and condom use beliefs among male prison inmates in two South African provinces: Mpumalanga and KwaZulu-Natal. AB - The purpose of this study was to describe the characteristics of a random sample of prison inmates from two provinces in South Africa with respect to correct knowledge regarding HIV/AIDS and sexual transmitted infection (STI) risk and beliefs about condom use. This cross-sectional descriptive study of 357 prison inmates formed part of a larger longitudinal investigation designed to implement a health education intervention for prison inmates in the KwaZulu-Natal (KZN) and Mpumalanga (MP) provinces of South Africa. Mean differences for groups were compared across sites using Analysis of Variance (ANOVA). Inmates from KZN were less likely to agree that 'It is important to use condoms every time you have sex' F (1355) = 25.8, p<0.0001 when compared with inmates in MP. However, they were more likely to agree that 'Condoms work well to prevent the spread of HIV' F (1355) = 11.7, p<0.001; 'Condoms also prevent pregnancy' F (1355) = 5.1, p<0.05. Overall, the demonstrated differences in condom use behaviour suggested that future prevention efforts focus on the importance of using condoms to prevent the spread of HIV/AIDS and other STIs. PMID- 19548158 TI - Drug resistance, patent resistance: Indian pharmaceuticals and the impact of a new patent regime. AB - This article highlights potential public health effects of India's Patents Act of 2005, which was implemented to conform to the requirements of the World Trade Organisation's Trade-Related Aspects of Intellectual Property Agreement (TRIPS), a new legal regime that will likely have a significant impact on access to HIV/AIDS medications in much of the world. This new patent law may play a role in keeping new antiretroviral (ARV) medications, including improved first-line medications and second-line drugs that are being developed for first-line drug resistant HIV, financially out of reach for many people living with HIV/AIDS in poor countries. India's drug industry, which had thrived under earlier patent laws that protected processes but not products in the case of medications, had brought down the price of ARV drugs in South Asia and Africa by more than 90%. While most existing drugs are grandfathered under the new patent laws, newer ARV medications may be barred from manufacture by Indian companies. This article analyses the effects of the coming together of this new legal regime, the global political economy and emerging resistance to HIV/AIDS medications, and evaluates efforts to mitigate the negative public health effects of the new patent laws. PMID- 19548159 TI - Sexing development and relocating gender: an organisational case study. AB - While gender has become increasingly mainstreamed in international development, it has lost its sharpness as an analytical tool and operational means to women's liberation. Sexuality, while it intersects with gender as a social construct, has been largely ignored within development contexts. This paper draws on findings that emerged from the gains and struggles experienced by project staff in a large international development agency engaging communities on issues related to sexual and women's rights. In so doing, it illustrates how a sexuality lens can move beyond gender equity and sexual health programmes focused on women's vulnerability (largely framed around protecting women and promoting access to services and contraceptive or condom use) to richer understandings of the range of social and cultural factors shaping sexual meanings and gender roles. Reshaping understandings of gender and sexuality requires a change from the conventional development discourse based on biomedical and moral-based sensibilities that associates sex with risk and danger, towards one that also recognises pleasure and agency. Sexuality is a vital aspect of development and sexual rights are a precondition not only for HIV/AIDS prevention, reproductive health or rights, but also for gender equality and social inclusion more widely. While personal and organisational transformation is fundamental, broader social, cultural and political forces, from community groups to governments and donors at large, must lie at the heart of change. PMID- 19548160 TI - Cigarette purchasing behaviour in Thailand and Malaysia: comparative analysis of a semi-monopolistic and a free-market structure. AB - A wide range of cigarette prices can undermine the impact of tobacco tax policy when smokers switch to cheaper cigarettes instead of quitting. In order to better understand this behaviour, we study socio-economic determinants of price/brand choices in two different markets: a semi-monopolistic market in Thailand and a competitive market in Malaysia. The hypothesis that the factors affecting the price/brand choice are different in these two markets is analysed by employing a 2005 survey among smokers. This survey provides a unique perspective on market characteristics usually described only in business reports by the tobacco industry. We found that smokers in Thailand have fewer opportunities to trade down to save money on cigarettes, but pay lower prices than smokers in Malaysia, despite Thailand's higher tax rate. The Malaysian market, on the other hand, offers many possibilities to shop around for cheaper cigarettes. Higher income and education increase the price paid per cigarette in both countries, but the impact of these factors is larger in Malaysia. This has implications for sensitivity to cigarette prices. Using tax policy alone should be a more effective tobacco control measure in Thailand as compared to Malaysia. The effectiveness of a tax increase in Malaysia can be improved by adding programmes focusing on smoking cessation among low-income/low-educated smokers. PMID- 19548162 TI - Kiwifruit-based polyphenols and related antioxidants for functional foods: kiwifruit extract-enhanced gluten-free bread. AB - The present study investigates the production of gluten-free bread enhanced with polyphenols and related antioxidants derived from a natural aqueous extract from green-fleshed kiwifruit (Actinidia deliciosa). Puree and four aqueous extracts, produced from ripe green kiwifruit in the absence of artificial preservatives, were subjected to storage stability trials at 4 degrees C, 20 degrees C and 38 degrees C, and were chemically characterized (phenolic, vitamin C and pectic polysaccharide contents). The aqueous extract with good stability and high phenolic and vitamin C contents was used for gluten-free bread-making. The resultant kiwifruit extract-enhanced bread was acceptable to a taste panel, possessing softer and smoother texture than plain gluten-free bread. Thus, the aqueous extract of kiwifruit puree containing health-beneficial constituents can be considered a functional ingredient for gluten-free bread formulation. PMID- 19548163 TI - Effect of flaxseed gum on reduction of blood glucose and cholesterol in type 2 diabetic patients. AB - The effects of ingestion of flaxseed gum on blood glucose and cholesterol, particularly low-density lipoprotein cholesterol, in type 2 diabetes were evaluated. Flaxseed gum was incorporated in wheat flour chapattis. Sixty patients of type 2 diabetes were fed a daily diet for 3 months, along with six wheat flour chapattis containing flaxseed gum (5 g), as per the recommendations of the American Diabetic Association. The control group (60 individuals) consumed an identical diet but the chapattis were without gum. The blood biochemistry profiles monitored before starting the study and at monthly intervals showed fasting blood sugar in the experimental group decreased from 154 +/- 8 mg/dl to 136 +/- 7 mg/dl (P=0.03) while the total cholesterol reduced from 182 +/- 11 mg/dl to 163 +/- 9 mg/dl (P=0.03). Results showed a decrease in low-density lipoprotein cholesterol from 110 +/- 8 mg/dl to 92 +/- 9 mg/dl (P=0.02). The study demonstrated the efficacy of flax gum in the blood biochemistry profiles of type 2 diabetes. PMID- 19548164 TI - Effectiveness of dietary intervention for obese women in Jordan. AB - OBJECTIVE: The aim was to evaluate the outcome of body weight loss consulting in an outpatient nutrition clinic. METHODS: Forty-five adult females attended 10 individualized treatment one-to-one sessions. Weight and height were measured and the body mass index was calculated. Triceps, biceps, subscapular and suprailiac skinfold thickness were measured and the fat percentage was calculated. A hypocaloric diet was given to the women; the percentages of carbohydrate, protein and fat in the diet were kept between 50 and 55% for carbohydrates, between 15 and 20% for protein, and < or =30% of fat. RESULTS: Average weight loss was 7.4 kg, which was 8.4% of initial. Class III obese subjects achieved the highest weight loss (-9.4 kg). Weight loss was statistically significant after week 1, week 2, week 3, and week 4 (P < 0.001). The body mass index was significantly decreased (P < 0.001). The mean fat percentage was not significantly different. CONCLUSIONS: The results indicate the increasing importance of nutrition counselling in Jordan. PMID- 19548165 TI - The contingency-shifting variant Iowa Gambling Task: an investigation with young adults. AB - The contingency-shifting variant Iowa Gambling Task (IGT), in which the reward and punishment contingencies of different decks of cards are systematically altered, was investigated with a large group of healthy young adults (n = 208). Our findings demonstrate that the onset of unsignaled, contingency-shift phases initially disrupted learning but that performance subsequently improved during each shift. Subjective experience ratings were positively correlated with performance across all phases. A regression model showed that performance early in the task, in Blocks 3 and 4, significantly predicted later ability to shift to the changing contingencies. Subdividing participants into high performer and low performer groups revealed an increased number of selections of previously good now-bad decks in the latter group. Overall, the contingency-shifting variant IGT may have potential as a novel measure of reversal learning in experimental and clinical settings. PMID- 19548166 TI - Alexithymia and emotional empathy following traumatic brain injury. AB - The frequency of alexithymia and the proportion of cases reporting low emotional empathy after traumatic brain injury (TBI) were compared with a control group. The study also examined the relationship between alexithymia and emotional empathy, controlling for the influence of cognitive ability, severity of head injury, and time since injury. A total of 64 TBI patients and matched controls completed the 20-Item Toronto Alexithymia Scale (TAS-20) and Balanced Emotional Empathy Scale (BEES). The TBI group exhibited a significantly higher frequency of alexithymia (60.9%) and low emotional empathy (64.4%) than did the control group (10.9% and 34.4%). Significant moderate negative correlations were found between TAS-20 and BEES scores, with TAS-20 total scores accounting for a significant amount of variance in BEES scores. However, no significant correlation was obtained between Subscale 1 of the TAS-20 (difficulty identifying feelings) and BEES scores in the TBI group. Additionally, there were no significant relationships between alexithymia, emotional empathy, injury severity, and time since injury. The results suggest an inverse relationship between alexithymia and emotional empathy. PMID- 19548167 TI - Impaired procedural learning in language impairment: results from probabilistic categorization. AB - The Weather Prediction (WP) Task is a classical task of probabilistic category learning generally used for examining the dissociation of procedural and declarative memory. The current study focuses on performance of children with language impairment (LI) and compares their performance to that of typically developing (TD) children and adults with the aim of testing the procedural deficit hypothesis of LI (PDH; Ullman & Pierpont, 2005), which states that language impairment is not a specific linguistic phenomenon, but results from the dysfunction of a more general cognitive system: the procedural system. To test the generality of the procedural impairment, we needed a task that is dissimilar from language in that it does not build on sequential information. Children with language impairment show deficient learning on the Weather Prediction Task, which already appears at the early stages of the task. These results, in line with the PDH, point to the deficit of the procedural system in language impairment going beyond the language system. Whether this deficit is selective to the procedural system or is complemented by deficits in the declarative system is the subject of future studies. PMID- 19548168 TI - Asymmetry in face processing during childhood measured with chimeric faces. AB - Studies with adults have found a left bias in their perception of faces, which suggests a right hemisphere specialisation in processing facial information. Hemispheric asymmetry is observed during the first year of life but it is still unclear at what age such specialisation becomes adult-like during the prolonged period of face-processing development through childhood. In the present study we investigated the development of children's perceptions of gender and emotion using chimeric faces. Our results demonstrate that a right hemisphere bias, similar to that found in adults, is observed from 5 years of age. The results are discussed within a framework of developmental studies and we conclude that although children may be less efficient than adults at encoding faces, their basic manner of encoding is not fundamentally different from that of adults. PMID- 19548169 TI - Personal computers in the voice laboratory: Part one--the computing environment. PMID- 19548171 TI - Efficacy of voriconazole in Japanese quail (Coturnix japonica) experimentally infected with Aspergillus fumigatus. AB - Aspergillus fumigatus causes disease in birds. Our objectives were to determine pharmacokinetic parameters and evaluate efficacy of voriconazole (VCZ) in a novel experimental quail model. After a single oral VCZ dose of 20 or 40 mg/kg, plasma concentrations peaked 2 h postdose (5.8 and 6.9 microg/ml) and remained above 0.5 microg/ml for 4 and 12 h postdose, respectively. For the efficacy study, ten-day old Japanese quail (n = 60) were infected intratracheally with A. fumigatus conidia. Daily oral VCZ at 20 or 40 mg/kg was initiated 24 h postinfection (PI); infected diluent-treated birds were given de-ionized water orally. Preassigned birds were euthanized on days 5 or 10 PI. VCZ at 40 mg/kg prolonged survival compared to 20 mg/kg or diluent-treatment (P < 0.05) and lungs from birds given VCZ at 40 mg/kg had fewer colony forming units (CFU) than diluent-treated (P = 0.03). At day 10 PI, birds treated with VCZ at 20 mg/kg had significantly fewer fungi in the lungs as demonstrated by methenamine silver stain (P = 0.017) or immunohistochemistry, as compared to diluent-treated (P = 0.034). Histopathologically, VCZ-treated birds did not have necrotic lesions and showed a trend toward fewer with acute inflammatory changes. VCZ at 40 mg/kg was efficacious in quail with experimental pulmonary aspergillosis. PMID- 19548172 TI - Prepartum autobiographical memory specificity predicts post-traumatic stress symptoms following complicated pregnancy. AB - Prior research has shown that reduced autobiographical memory specificity predicts an increase in post-traumatic stress severity in traumatised individuals. Studies have also demonstrated that reduced memory specificity predicts later symptoms of depression after pregnancy-related life stress. So far, no reported studies have tested the predictive value of memory specificity at the onset of a potentially traumatic situation. Therefore the aim of the present study was to investigate whether prenatal memory specificity would predict post-traumatic stress after complicated pregnancy. The results demonstrate that women who retrieved fewer specific memories with a pregnancy related content to positive cues during pregnancy (i.e., directly after hospitalisation) reported more post-traumatic stress 6 weeks after giving birth. This relationship remained significant after controlling for variables that were related to both baseline autobiographical memory specificity and later post traumatic stress. A similar pattern was found for depression symptomatology, even when somatic symptoms were excluded from the analyses. Taken together, these data suggest that the relationship of memory specificity with later depression can be generalised to post-traumatic stress symptoms. PMID- 19548173 TI - Familiar and unfamiliar face recognition: a review. AB - Since the 1970s there has been a continuing interest in how people recognise familiar faces (Bruce, 1979; Ellis, 1975). This work has complemented investigations of how unfamiliar faces are processed and the findings from these two strands of research have given rise to accounts that propose qualitatively different forms of representation for familiar and unfamiliar faces. Evidence to suggest that we process familiar and unfamiliar faces in different ways is available from cognitive neuropsychology, brain scanning, and psychophysics. However, in this review we focus on the evidence, available from experimental investigations of how people recognise faces, for different types of representation existing for each type of face. Factors affecting recognition are evaluated in terms of how they apply to familiar and unfamiliar faces and categorised according to the nature of their impact. In the final section this evidence, along with recent advances in the field, is used to explore the way in which unfamiliar faces may become familiar and the factors that may be important for the development of familiar face representations. PMID- 19548174 TI - Memory conjunction errors: the effects of presentation duration and study repetition. AB - The memory-strengthening manipulations of increased presentation duration and increased number of times items were presented were manipulated in the memory conjunction paradigm. Participants viewed parent words once or three times during the study portion of the experiment for 250 ms, 1000 ms, or 3000 ms. After an old/new recognition test participants were asked to give explanations for their answers from the recognition test. The results of true and false recognition as well as recall-to-reject responses (e.g., I know I did not see blackbird since I saw blackmail) indicated that both familiarity and recollection were influenced by the memory-strengthening manipulations. The results provide evidence for dual process theories of recognition memory and the opposing processes of familiarity and recollection. PMID- 19548175 TI - Potential applications of medical and non-medical robots for neurosurgical applications. AB - The objective of the paper is to review the state-of-the-art in medical robotic systems used for different surgical applications, and to position and evaluate their concepts according to the design requirements of an innovative, robotized neurosurgical system, capable of performing tumor ablation or electrode positioning. A few other non-medical systems, which have interesting concepts, will also be discussed. The overall aim is to determine the robotic concept (structure, actuation, etc.) most applicable to specific tasks in neurosurgery. The first section of the article describes the requirements of the task and each important aspect is expressed by an evaluation criterion. Then, 59 systems are described, according to the fields of medical applications and the robotic concepts. An evaluation of the different systems is conducted, based on the five most significant criteria. However, the main characteristic assessed is the deployment capability of the system i.e. extension and retraction. The final section presents an overview of concepts transferable to neurosurgical applications. Continuum concepts, such as "elephant trunks", seem to be the most adapted solutions, utilizing pneumatic and/or spring and/or cable actuations. Pneumatics offer deployment forces and cables can control and guide the deployment. The design of a new neurosurgical device should take into account these observations as a base. PMID- 19548176 TI - Directions in brain injury research: From concept to clinical implementation. AB - Development of rehabilitation treatments and services that are effective and supported by rigorous evidence is a long and multi-step process. Although there are similarities between this process and that used in pharmaceutical development, the sequence involved in developing this evidence in rehabilitation is more complex and less linear. The specific steps involved depend on a variety of factors including the location of the target of treatment within the International Classification of Functioning, Disability, and Health (ICF; World Health Organisation, 2006), the degree to which the treatment is guided by an underlying theory, the availability of animal models, and the plausibility of safety concerns. Similarly, the research designs chosen and the setting of the research vary both by treatment target and by maturity of the programme of evidence development. A full picture of the appropriate role and anticipated impact of specific rehabilitation treatments can benefit from a marriage between rehabilitation research (which seeks to understand how to change specific aspects of function), and enablement/disablement research (which seeks to understand how changes in one part of the ICF framework affect functioning elsewhere). Although the specific steps involved in a programmatic approach to rehabilitation treatment research vary, development of evidence about all treatments can benefit from a systematic programmatic approach in which each step seeks to answer specific questions and set the stage for subsequent investigation. PMID- 19548177 TI - Screening of psychiatric outpatients for borderline personality disorder with the McLean Screening Instrument for Borderline Personality Disorder (MSI-BPD). AB - BACKGROUND: Detecting patients with borderline personality disorder (BPD) is important, and feasible screening instruments are needed. AIMS: To investigate our Finnish translation of the McLean Screening Instrument for Borderline Personality Disorder (MSI-BPD) as a screen for BPD among psychiatric outpatients, its psychometric and screening properties, and feasibility in improving the recognition of BPD. METHODS: We screened 302 consecutive psychiatric outpatients at the Department of Psychiatry at the Helsinki University Central Hospital in Finland for BPD using the Finnish MSI-BPD. Of the patients, 69 (23%) were assigned to a random sample that was stratified according to the number of screens returned to the outpatient clinics, and further stratified into the three strata, high scores deliberately enriched, according to the MSI-BPD scores. Finally, a stratified random sample of 45 patients was interviewed with the Structured Clinical Interview for DSM-IV Axis II Personality Disorders (SCID-II) by the interviewers blind to the patients' MSI-BPD scores. RESULTS: One third (29%) of 302 screened patients had a positive MSI-BPD. The internal consistency of the MSI-BPD was good (Cronbach's alpha = 0.77). Of the 45 patients interviewed with the SCID-II, 11 (24%) were found to have BPD, five (46%) of whom a previously clinical diagnosis. In a ROC analysis, the optimal cut-off score was 7. CONCLUSIONS: The translated MSI-BPD was found to be a feasible screen for BPD in Finnish psychiatric outpatient care. Further studies investigating the clinical utility of MSI-BPD in larger clinical samples are warranted. PMID- 19548178 TI - Comparison of platelet aggregation using light transmission and multiple electrode aggregometry in Glanzmann thrombasthenia. AB - We examined platelet aggregation in platelet-rich plasma (PRP) and in whole blood in nine patients with Thrombasthenia Glanzmann (TG). In PRP, aggregation was measured by monitoring the changes in light absorbance that occurred in response to adenosine 5-diphosphate (ADP), collagen and ristocetin. To measure platelet aggregation in whole blood, we used multiple electrode Impedance aggregometry using the same aggregating agents. In PRP, the patient's platelets showed defective aggregation in response to ADP, collagen, epinephrine and partially to ristocetin in all patients. In whole blood, platelet aggregation in response to the same aggregating agents showed similar response and appeared to be very similar to that which occurred in PRP. Whole blood impedance aggregometry seems to give similar results to PRP light transmission aggregometry in patients with TG. Multiple electrode aggregometry (MEA) is faster and more convenient to use in these patients. PMID- 19548179 TI - General practice needs to improve recruitment and income of GP researchers. PMID- 19548181 TI - Thalamic contributions to anterograde, retrograde, and implicit memory: a case study. AB - Learning and memory deficits are typically associated with damage or dysfunction of medial temporal lobe structures; however, diencephalic lesions are another common cause of severe and persistent memory deficits. We focus specifically on the thalamus and review the pathological and neuropsychological characteristics of two common causes of such damage: Korsakoff's syndrome and stroke. We then present a patient who had sustained bilateral medial thalamic infarctions that affected the medial dorsal nucleus and internal medullary lamina. This patient demonstrated the characteristic temporally graded retrograde amnesia and a profound anterograde memory (i.e., explicit memory) deficit within the context of relatively preserved implicit memory. Implications of this explicit-implicit discrepancy are discussed within the context of cognitive rehabilitation techniques that hold promise for more severely impaired patients. PMID- 19548182 TI - Late-onset obsessive compulsive disorder associated with possible gliomatosis cerebri. AB - Onset of obsessive-compulsive disorder (OCD) after the age of 50 years is rare, and should alert the physician to possible "organic" causes of OCD. These include infections, degenerative disorders, brain injury and cerebrovascular lesions, principally involving the frontal lobes and basal ganglia. The current patient had obsessive images, anxiety, auditory hallucinations and seizures following (possible) gliomatosis cerebri, with onset around 69 years of age. The atypical presentation, lesions involving the cortical-basal ganglia-thalamic-cortical circuit and the association with neurological signs/symptoms, was characteristic. However, late-onset OCD has not been commonly reported with diffuse lesions, and the association with gliomatosis cerebri is not known. This patient's case illustrates the need for careful screening of older patients with recently acquired OCD, and for further systematic study of OCD in the broad range of neuropsychiatric disorders affecting the elderly. PMID- 19548184 TI - Tumor size of Conn's adenoma and comorbidities. AB - Primary aldosteronism (PA) is associated with vascular end organ damage. The aim of the study was to evaluate differences regarding comorbidities depending on tumor size in patients with aldosterone producing adenoma (APA). The retrospective cross-sectional study was done by collection from 6 German centers (German Conn's registry) between 1990 and 2007. Among the 640 registered patients with PA, 60 operated patients with APA were analyzed. The main outcome of measures was the comorbidities depending on tumor size. Thirty-one patients (17 men, 14 women) had an adenoma size <20 mm, and 29 patients (10 men, 19 women) had an adenoma size>/=20 mm. There was no difference in age, preoperative potassium, aldosterone, or creatinine levels, preoperative systolic and diastolic blood pressure, or duration of hypertension between the two groups. In the group with APA <20 mm, cerebrovascular events occurred with a prevalence of 12.9%, cardiac events 16.1%, peripheral vascular events 25.8%, renal insufficiency 16.1%, and sleep apnea 6.4%, respectively. There was no significant difference in comorbidities compared to the group with APA>/=20 mm. Subgroup analysis (n=22) of follow-up data on post-operative systolic and diastolic blood pressure showed no significant difference between these subgroups with regard to potassium, aldosterone or creatinine levels, blood pressure, duration of hypertension, or comorbidities. Our data indicate a high prevalence of comorbidities in patients with APA. However, adenoma size was not correlated with cardio- and cerebrovascular comorbidities, and does not seem to be a prognostic factor for blood pressure outcome. PMID- 19548185 TI - [Modular semiquantitative quality assessment of ophthalmic health information on the internet--reproducibility and correlation between different assessment categories]. AB - BACKGROUND: Quality evaluation of web-based health information in ophthalmology requires valid standards and reproducible assessment procedures. The objective was to evaluate the interrater-reliability of quality assessment of ophthalmic websites and a possible correlation between the results of the evaluation categories reliability/trustworthiness, quality of medical content and accessibility/usability. MATERIALS AND METHOD: After selection with the search engine "Google" 20 ophthalmic websites had been analysed by two independent evaluators using criteria checklists (modified Afgis transparency criteria, modified BITV test, medical content related to AMD) related to the aforementioned 3 categories. RESULTS: The interrater-reliability was almost perfectly estimated with Kappa-values of 0.91 for reliability/trustworthiness plus 0.89 for accessibility/usability and 0.79 for content. On average 62.5 % (+/- 17.43 %), 27.36 % (+/- 16.5 %) and 59.54 % (+/- 15.73 %) of the quality requirements were fulfilled for reliability/trustworthiness, content and accessibility/usability, respectively. No significant correlation was found between reliability and content (r = -0.039, p = 0.8709), reliability and accessibility/usability (r = 0.284; p = 0.228) plus content and accessibility/usability (r = 0.199; p = 0.4047). CONCLUSION: Sufficiently operationalised criteria are prerequisites for reproducible results of quality assessment of ophthalmic websites between different observers. The assessment within a single category, such as reliability/trustworthiness, does not allow one to draw conclusions on other categories such as content or accessibility/usability or the overall quality of a website. Therefore, simplified tools for quality assessment of health information by laymen and patients may have a limited validity. PMID- 19548186 TI - [Measurement set-up for estimation of colour discrimination thresholds of colour vision deficiencies and while wearing coloured filters]. AB - BACKGROUND: In clinical practice, several techniques for the evaluation of colour vision deficiencies are well established. Most of them are designed for the detection of congenital or pathological changes. Due to the narrow-banded light sources, used in monitors or anomaloscopes, the human gamut can be addressed only in part. With these set-ups, the impact of coloured filters on colour vision such as blue light filtering intraocular lenses cannot be investigated precisely. MATERIAL AND METHODS: The technique is based on the visual matching of colour differences between two halves of a test field. The colours are generated by mixing seven types of LEDs. A transparent, vertically divided circular test field is retro-illuminated with this light using optical fibres and projection optics. An ocular optic assures the unaccommodated sight of the subject on the test field (size 2 degrees). RESULTS: The developed set-up presents freely adjustable colours in the form of continuous light spectra to examine the influence of filter materials on colour discrimination. A colour space can be generated which corresponds to that of human perception. The saturation and brightness of the light is adjustable. The subject has to decide if he can discriminate between the colours in the test field halves or not. Before the next colours are shown the subject is neutrally adapted by a neutral hue to provide different adaptation levels. The luminescence of the test field can be varied up to 1500 cd/m(2). The adaptive algorithm of the sequence is based on the SIAM method described by Kaernbach. Based on reference measurements, including 4 subjects with a congenital deficiency, it was shown that the discrimination thresholds at the characteristic confusing axes are significantly different than those with normal colour vision. Also, the impact of green laser goggles on the colour perception was determined. Due to the filter the thresholds in the vicinity of yellow (x = 0.4664; y = 0.4525) are reduced in the yellow-orange and cyan range and increased in the green-yellow and magenta range. CONCLUSION: With this set-up an objective instrument is available for clinical research in the field of individual human colour perception. It enables an accurate and detailed analysis of the impact of coloured filters as well as a quantitative detection of congenital anomalies. The adaptive algorithm allows short sequences and precise determination of colour discrimination thresholds. PMID- 19548187 TI - Protective effect of acteoside on immunological liver injury induced by Bacillus Calmette-Guerin plus lipopolysaccharide. AB - The hepatoprotective effects of acteoside from O. coerulescens were evaluated in BCG plus LPS-induced immunological liver injury (ILI) in mice. Acteoside (50, 150, or 300 mg/kg) was administered via gavage daily for 12 days. The liver index (liver weight/body weight), liver homogenate levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), hepatic nitric oxide (NO), malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, production of tumor necrosis factor-gamma (TNF-gamma) and interleukin-2, 4, 10 (IL-2, 4, 10), as well as histopathological changes of the liver were evaluated following the 12 day treatment. Moreover, the modulation influence of acteoside on the expression of B cell lymphoma/leukemia-2 (Bcl-2, hepatocyte apoptosis inhibitor) and Bcl-2 associated X protein (Bax, hepatocyte apoptosis promoter) in the mice liver with immunological hepatic injury was studied also. Acteoside (50, 150, or 300 mg/kg) effectively reduced the BCG/LPS-induced elevated liver index, liver homogenate AST and ALT levels, hepatic NO and MDA contents, restored hepatic SOD activity and reduced the degree of liver injury in ILI mice. The expression of Bax was decreased (vs. BCG + LPS model group), while the expression of Bcl-2 increased (vs. BCG + LPS model group). These results are close to those of DDB (as a reference drug), and suggest that acteoside has a protective and therapeutic effect on ILI mice, which might be associated with its antioxidant properties, immunoregulatory function and regulation of hepatic apoptosis. PMID- 19548188 TI - Secondary metabolic profiling and artemisinin biosynthesis of two genotypes of Artemisia annua. AB - Artemisinin has been proven to be an effective antimalarial compound, especially for chloroquine-resistant and cerebral malaria. However, its biosynthesis pathway is still not completely clear. In order to get new clues about artemisinin biosynthesis, metabolic profiling by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) was applied to compare the secondary metabolites of two Artemisia annua L., genotype SP18 and 001, for some phenotypic and agricultural trait differences, including artemisinin content, existed between the two genotypes. Samples at 7 time points of three growth stages were studied. The data of profiles were subjected to multivariate analysis with partial least squares discriminant analysis (PLS-DA). The results indicated that there were clear differences in terpenoids and artemisinin metabolism between different growth stages and genotypes. Twenty-one compounds, including artemisinin and its related precursors, were selected as the marker compounds of the PLS-DA between the two genotypes. Among them, artemisinic acid, arteannuin B, borneol, beta-farnesene and an unidentified sesquiterpenoid (peak 48) were abundant in 001, while camphor, methyl artemisinic acid and lanceol accumulated mainly in SP18. The relationship between these differences and artemisinin biosynthesis in the two genotypes of A. annua were discussed. PMID- 19548189 TI - Protective effects of echinacoside, one of the phenylethanoid glycosides, on H(2)O(2)-induced cytotoxicity in PC12 cells. AB - We have investigated the protective effects of echinacoside (ECH), one of the phenylethanoid glycosides, on H(2)O(2)-induced cytotoxicity in the rat pheochromocytoma cell line (PC12 cells). Our data show that application of ECH to H(2)O(2)-injured PC12 cells (HIPCs) increased cell viability and decreased the apoptotic ratio. Flow cytometry (FCM) and laser scanning confocal microscopy (LSCM) analysis suggested that ECH exerted its inhibitory effects on the formation of reactive oxygen species (ROS) and the accumulation of intracellular free Ca(2+) ([Ca(2+)]i). In addition, ECH elevated the mitochondrial membrane potential (MMP) in HIPCs. Furthermore, Western blot analysis revealed that ECH prevented an H(2)O(2)-induced increase of the Bax/Bcl-2 ratio by down-regulating Bax protein expression and upregulating Bcl-2 protein expression. In summary, ECH showed significant neuroprotective effects on HIPCs through the mitochondrial apoptotic pathway, and could be a potential candidate for intervention in neurodegenerative diseases such as Alzheimer's and Parkinson's disease. PMID- 19548190 TI - Changes of gentiopicroside synthesis during somatic embryogenesis in Gentiana macrophylla. AB - IN VITRO plant regeneration of Gentiana macrophylla Pall. and determination of gentiopicroside content during somatic embryogenesis are described in the present work. The highest percentage of embryogenic callus formation was observed in Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4 dichlorophenoxyacetic acid (2,4-D) and 1.0 mg/L 6-benzylaminopurine (BA). Calli were subcultured on MS medium containing 1.0 mg/L 2,4-D, 1.0 mg/L BA and 500 mg/L lactalbumin hydrolysate (LH) at intervals of 25 days. A higher frequency of somatic embryo maturation was achieved on MS medium containing B5 vitamins (MB) supplemented with different concentrations of 1-naphthaleneacetic acid (NAA) and BA than with a combination of NAA and kinetin (KT). Addition of AgNO(3) improved maturation of somatic embryos while thidiazuron (TDZ) promoted vitrification. The gentiopicroside contents of embryogenic calli and globular-, heart-, torpedo-, and cotyledon-shaped embryoids were determined by high-performance liquid chromatography (HPLC). Gentiopicroside was not detectable in embryogenic calli, but in all types of somatic embryos. The highest gentiopicroside content was observed in cotyledon-shaped embryoids, reaching more than 12 mg/g dry weight. PMID- 19548191 TI - Anti-ischemic activity and endothelium-dependent vasorelaxant effect of hydrolysable tannins from the leaves of Rhus coriaria (Sumac) in isolated rabbit heart and thoracic aorta. AB - The aim of this work was to investigate the cardioprotective activity of hydrolysable gallotannins from Rhus coriaria L. leaves extract (RCLE) in isolated rabbit heart preparations, submitted to low-flow ischemia/reperfusion damage. RCLE induces a dose-dependent normalization of coronary perfusion pressure (CPP), reducing left ventricular contracture during ischemia, and improving left ventricular developed pressure and the maximum rate of rise and fall of left ventricular pressure at reperfusion. Creatinine kinase (CK) and lactate dehydrogenase (LDH) outflow were significantly reduced during reperfusion. In parallel there was a rise in the release of the cytoprotective 6 ketoprostaglandin F (1alpha) (6-keto-PGF (1alpha)) and a decrease of tumor necrosis factor-alpha (TNF-alpha), both significant only at the highest RCLE concentrations (150-500 microg/mL). The vasorelaxant activity of RCLE was studied in isolated rabbit aorta rings precontracted with norepinephrine (NE) with and without endothelium. The vasorelaxation induced by RCLE was predominantly endothelium-dependent as demonstrated by the loss of RCLE vasorelaxant ability in i) de-endothelized rings and ii) in intact aortic rings after pretreatment with NG-monomethyl- L-arginine (L-NMMA) and 1 H-[1.2.4]oxadiazolo[4.3- A]quinoxalin-1 one (ODQ). The inhibition of vasorelaxation in intact rings by indomethacin (INDO) demonstrates the ability of RCLE to modulate the coronary endothelium cyclooxygenase (COX) pathway. The K-ATP channel antagonist glibenclamide (GLIB) was ineffective. The antioxidant activity of RCLE, investigated in the 1,1 diphenyl-2-picrylhydrazyl (DPPH) model and in living cell systems (rat erythrocytes), was stronger than that of gallic acid, ascorbic acid and trolox. The structure of its main bioactive constituents, profiled by HPLC-ESI-HR-S, comprised a mixture of polygalloylated D-glucopyranose with different degrees of galloylation and 3- O-methylgallic acid. The cardiovascular protective effect of RCLE seems to be due to an interplay of different factors: COX pathway activation, TNF-alpha inhibition, endothelial nitric oxide synthase (eNOS) activation, and free radical and ROS scavenging. PMID- 19548192 TI - The phytohormone auxin induces G1 cell-cycle arrest of human tumor cells. AB - The plant hormone auxin is the key regulator of plant growth and development. Auxin regulates transcription of plant genes by targeting degradation of transcriptional repressor proteins Aux/IAA. While there are many reports describing its potential to modulate human cell functions, the majority are based on auxin action following enzymatic activation. A study focused on auxin alone and its antiproliferative potential, with emphasis on modulation of the cell cycle, has not been performed. Therefore, we analyzed tumor growth inhibitory effects and the cell-cycle perturbations of natural (IAA, IBA) and synthetic (NAA, 2,4-D) auxins. All derivatives showed cytostatic effects on selected human tumor cell lines. The cell-cycle analysis revealed that IAA and 2,4-D induce strong G1 arrest, along with a drastic decrease in the percentage of S-phase cells in MCF-7 cell line. This phenomenon demonstrates that auxins may have novel, unexploited antitumor potential and should be further investigated. PMID- 19548193 TI - Changes in the epidemiology of acute appendicitis and appendectomy in Danish children 1996-2004. AB - PURPOSE: Aim of the study was to describe changes in the epidemiology of acute appendicitis in Danish children between 0-19 years of age for the period 1996 2004. METHODS: The study was based on discharge diagnoses taken from the Danish National Patient Registry of all 28 274 patients with a diagnosis of acute uncomplicated or complicated appendicitis, and/or a registered procedure code of appendectomy. These data were computed together with data on the background population, and incidences were calculated. RESULTS: A significant decrease in the incidence of acute uncomplicated appendicitis was found for all age groups (range, 13-36%). The decrease was present for both sexes, but most prominent in girls. The incidence of complicated acute appendicitis decreased by 10%. CONCLUSION: The incidence of acute appendicitis is declining. The incidence of uncomplicated appendicitis appears to be declining more rapidly than that of complicated appendicitis. This finding does not necessarily imply poorer management of the disease, but-as suggested by others-could be due to different behaviours of uncomplicated and complicated appendicitis. PMID- 19548194 TI - Patients with anorectal malformation and Hirschsprung's disease. AB - BACKGROUND: Dysganglionosis and aganglionosis have been frequently described in biopsies of the distal bowel in patients with anorectal malformation (ARM). They are interpreted as a developmental disorder of the anorectum. The true association of total colonic aganglionosis and anorectal malformation has not been reported before. The aim of this study was to explore the true association of anorectal malformation and Hirschsprung's disease with or without trisomy 21. MATERIAL & METHODS: A retrospective study of all patients diagnosed with anorectal malformation in our institute from 1986-2008 was performed. All patients with anorectal malformation and Hirschsprung's disease were included in the study. Rectal biopsies were taken from multiple sites, including the rectum, left, transverse, right colon and appendix. The diagnosis of aganglionosis was proven histopathologically by the absence of ganglion cells with or without acetylcholinesterase staining. Specimens were examined by at least two experienced consultant pathologists. RESULTS: Aganglionosis was confirmed in three patients out of 53 patients with anorectal malformation. Two had Down's syndrome. All were males and presented with high anorectal malformation without fistula. The clinical presentation was intestinal obstruction, necrotizing enterocolitis and failure to thrive. The level of aganglionosis was up to the left colon in two and total colonic with ileal involvement in one. One of the children with Down's syndrome and total colonic aganglionosis died. Another had correction of a congenital heart disease, colostomy and is awaiting definitive surgery. The third case is continent at the age of 22 years with a Malone stoma after pull-through of ARM and a subsequent Duhamel procedure. CONCLUSIONS: The association between ARM and intestinal dysganglionosis is not rare. We recommend not using the distal rectal pouch and parts of the fistula in the reconstruction of anorectal malformations as this may solve the constipation if the pathology is limited. In cases of aganglionosis beyond the rectal pouch and fistula, surgical intervention is needed. Delay in diagnosis may lead to morbidity or even mortality. PMID- 19548195 TI - Wandering spleen causing intestinal obstruction after repair of congenital diaphragmatic hernia. PMID- 19548196 TI - Solid pseudopapillary tumor of the pancreas: raising awareness of a not so rare pancreatic tumor affecting young females. PMID- 19548197 TI - The evolution of biliary atresia in early life. PMID- 19548198 TI - The outcome of closing gastroschisis: two case reports [corrected] and literature review. PMID- 19548203 TI - Alcohol consumption and body weight. AB - The number of Americans who are overweight or obese has reached epidemic proportions. Elevated weight is associated with health problems and increased medical expenditures. This paper analyzes Waves 1 and 2 of the National Epidemiological Survey of Alcohol and Related Conditions to investigate the role of alcohol consumption in weight gain. Alcohol is not only an addictive substance but also a high-calorie beverage that can interfere with metabolic function and cognitive processes. Because men and women differ in the type and amount of alcohol they consume, in the biological effects they experience as a result of alcohol consumption, and in the consequences they face as a result of obesity, we expect our results to differ by gender. We use first-difference models of body mass index (BMI) and alcohol consumption (frequency and intensity) to control for time-invariant unobservable factors that may influence changes in both alcohol use and weight status. Increasing frequency and intensity of alcohol use is associated with statistically significant yet quantitatively small weight gain for men but not for women. Moreover, the first-difference results are much smaller in magnitude and sometimes different in sign compared with the benchmark pooled cross-sectional estimates. PMID- 19548204 TI - Prenatal sonographic diagnosis of skeletal dysplasias. AB - OBJECTIVE: To assess the types and numbers of cases, gestational age at specific prenatal diagnosis and diagnostic accuracy of the diagnosis of skeletal dysplasias in a prenatal population from a single tertiary center. METHODS: This was a retrospective database review of type, prenatal and definitive postnatal diagnoses and gestational age at specific prenatal diagnosis of all cases of skeletal dysplasias from a mixed referral and screening population between 1985 and 2007. Prenatal diagnoses were grouped into 'correct ultrasound diagnosis' (complete concordance with postnatal pediatric or pathological findings) or 'partially correct ultrasound diagnosis' (skeletal dysplasias found postnatally to be a different one from that diagnosed prenatally). RESULTS: We included 178 fetuses in this study, of which 176 had a prenatal ultrasound diagnosis of 'skeletal dysplasia'. In 160 cases the prenatal diagnosis of a skeletal dysplasia was confirmed; two cases with skeletal dysplasias identified postnatally had not been diagnosed prenatally, giving 162 fetuses with skeletal dysplasias in total. There were 23 different classifiable types of skeletal dysplasia. The specific diagnoses based on prenatal ultrasound examination alone were correct in 110/162 (67.9%) cases and partially correct in 50/162 (30.9%) cases, (160/162 overall, 98.8%). In 16 cases, skeletal dysplasia was diagnosed prenatally, but was not confirmed postnatally (n = 12 false positives) or the case was lost to follow-up (n = 4). The following skeletal dysplasias were recorded: thanatophoric dysplasia (35 diagnosed correctly prenatally of 40 overall), osteogenesis imperfecta (lethal and non-lethal, 31/35), short-rib dysplasias (5/10), chondroectodermal dysplasia Ellis-van Creveld (4/9), achondroplasia (7/9), achondrogenesis (7/8), campomelic dysplasia (6/8), asphyxiating thoracic dysplasia Jeune (3/7), hypochondrogenesis (1/6), diastrophic dysplasia (2/5), chondrodysplasia punctata (2/2), hypophosphatasia (0/2) as well as a further 7/21 cases with rare or unclassifiable skeletal dysplasias. CONCLUSION: Prenatal diagnosis of skeletal dysplasias can present a considerable diagnostic challenge. However, a meticulous sonographic examination yields high overall detection. In the two most common disorders, thanatophoric dysplasia and osteogenesis imperfecta (25% and 22% of all cases, respectively), typical sonomorphology accounts for the high rates of completely correct prenatal diagnosis (88% and 89%, respectively) at the first diagnostic examination. PMID- 19548205 TI - Expression of toll-like receptor 9 in horse lungs. AB - Toll-like receptor 9 (TLR9) has been found to be the main receptor to respond to bacterial DNA in a wide variety of species. Recent work has shown that TLR9 is expressed in a diverse set of cells within the lung. However, much of this data has been centered on human and mouse cell culture lines or primary cultures and very little is known of TLR9 expression in intact lung, especially that of the horse. Here we show that TLR9 is expressed in the lungs of horses in a wide variety of cells. In particular, we note expression in pulmonary intravascular macrophages (PIMs), alveolar macrophages, bronchial epithelial cells, and type-II cells amongst others. Immunogold electron microscopy localized TLR9 in nuclei, cytoplasm, and plasma membrane of various lung cells. The data also show that E. coli lipopolysaccharide significantly increased expression of TLR9 mRNA in lungs and the number of cells in the lung septa that were positive for TLR9 protein. Protein expression was seen in airway epithelium, vascular endothelium, and inflammatory cells in blood vessels. Intravenous administration of gadolinium chloride, which depletes macrophages, before the lipopolysaccharide treatment significantly inhibited the LPS-induced increase in TLR9 mRNA in the lungs of the horses. We conclude that TLR9 is expressed in lung cells including PIMs and that the lipopolysaccharide treatment increases TLR9 mRNA expression. The increase in TLR9 mRNA is eliminated by depletion of PIMs, implicating these cells as a major source of TLR9 in the equine lung. PMID- 19548206 TI - Downregulation of pro-inflammatory cytokines by lupeol measured using cytometric bead array immunoassay. AB - The objective of the study was to investigate the activity of Lupeol (LUP) on proinflammatory and anti-inflammatory cytokines in the pleural exudate from male swiss albino mice. We applied Cytometric bead array technology for simultaneously measurement of these cytokines in pleurisy induced mice treated with lupeol in graded oral doses. Cytometric bead array uses the sensitivity of amplified fluorescence detection by flowcytometer to measure soluble analytes in a particle based immune assay. This assay can accurately quantitate 5 cytokines in a 50 microlitre sample volume. Oral administration of LUP at doses of 25, 50, 100 and 200 mg/kg p.o. produced dose related inhibition of IL-2, IFN-gamma and TNF-alpha in the pleural exudate with the most significant effect at 100 mg/kg oral dose. LUP had a non significant inhibitory effect on the levels of IL-4 and IL-5. PMID- 19548207 TI - Serum levels of brain-derived neurotrophic factor in comorbidity of depression and alcohol dependence. AB - Alcohol dependence is often comorbid with depression. The purpose of the present study was to compare serum brain-derived neurotrophic factor (BDNF) levels between depressive patients with and without alcohol dependence. Our subjects were 16 inpatients (M/F: 13/3, age: 48 +/- 8 years) at our university hospital who met the DSM-IV-TR criteria for both major depressive disorder and alcohol dependence and whose Hamilton Rating Scale for Depression (HAM-D) scores were at least 15. Twenty sex- and age-matched depressive patients and 20 healthy subjects were also examined. Serum BDNF levels in the depressive patients with (9.0 +/- 4.3 ng/ml) and without (9.8 +/- 5.2 ng/ml) alcohol dependence were significantly lower than those in the healthy subjects (21.1 +/- 7.0 ng/ml); however, no significant difference was found in the serum BDNF levels of depressive patients with and without alcohol dependence. Eight of the 16 (50%) depressive patients suffering from both depression and alcohol dependence responded to 8 weeks of treatment with antidepressant drugs which significantly increased their serum BDNF levels. These results suggest that the serum BDNF level is a useful biological marker for depression in patients with alcohol dependence. PMID- 19548208 TI - Automation in high-content flow cytometry screening. AB - High-content flow cytometric screening (FC-HCS) is a 21st Century technology that combines robotic fluid handling, flow cytometric instrumentation, and bioinformatics software, so that relatively large numbers of flow cytometric samples can be processed and analysed in a short period of time. We revisit a recent application of FC-HCS to the problem of cellular signature definition for acute graft-versus-host-disease. Our focus is on automation of the data processing steps using recent advances in statistical methodology. We demonstrate that effective results, on par with those obtained via manual processing, can be achieved using our automatic techniques. Such automation of FC-HCS has the potential to drastically improve diagnosis and biomarker identification. PMID- 19548210 TI - The lipophilicity indices of flavonoids estimated by reversed-phase liquid chromatography using different computation methods. AB - The chromatographic behavior of some flavonoids was established by RP-HPLC on RP 18 (LiChroCART, LiChrosphere RP-18e), RP-8 (Zorbax, Eclipse XDB-C8), CN (Saulentechnik, LiChrosphere CN100) columns. The mobile phases were mixtures of methanol-water in different volume proportions from 70 to 80% v/v for RP-18 and RP-8, while for the CN column the proportions were between 66 and 70% v/v. The lipophilicity was expressed through different lipophilicity descriptors such as mean of k (mk), mean of log k (mlog k), log k(W), S, f() and scores of k and log k corresponding to the first principal component. The experimental lipophilicity indices are directly correlated with the computed values, via computer software and internet module, at a high analytical level. Furthermore, the results obtained applying principal component analysis to k and/or log k values allow the prediction and explanation of the interaction involved in the retention mechanism which takes place between the compounds and the employed stationary phases during the development. PMID- 19548211 TI - Side-chain modified bile acids: chromatographic separation of 23-methyl epimers. AB - Owing to the re-flourished interest towards the bile acids (BAs) as versatile signalling hormones endowed with diverse endocrine functions, the development of reliable analytical protocols monitoring the synthesis of new BA-based receptor modulators, still represents a cogent concern. On this basis, for the first time, a HPLC study has been engaged with the aim to set up suitable chromatographic conditions for the analysis of three different epimeric couples of 23-methyl substituted unconjugated BAs. Three different methods (one for each couple) have been successfully established and then validated. Good precision and accuracy (evaluated both in the short and long period) as well as appreciably low LOD and LOQ values have turned out. Moreover, the engagement of an evaporative light scattering detector (ELSD) has proven its high effectiveness for the analysis of such steroidal species. PMID- 19548209 TI - Trajectories of mobility and IADL function in older patients diagnosed with major depression. AB - OBJECTIVE: Research has shown an association between depression and functional limitations in older adults. Our aim was to explore the latent traits of trajectories of limitations in mobility and instrumental activities of daily living (IADL) tasks in a sample of older adults diagnosed with major depression. METHODS: Participants were 248 patients enrolled in a naturalistic depression treatment study. Mobility/IADL tasks included walking one-fourth mile, going up/down stairs, getting around the neighborhood, shopping, handling money, taking care of children, cleaning house, preparing meals and doing yardwork/gardening. Latent class trajectory analysis was used to identify classes of mobility/IADL function over a 4-year period. Class membership was then used to predict functional status over time. RESULTS: Using time as the only predictor, three latent class trajectories were identified: (1) Patients with few mobility/IADL limitations (42%), (2) Patients with considerable mobility/IADL limitations (37%) and (3) Patients with basically no limitations (21%). The classes differed primarily in their initial functional status, with some immediate improvement followed by no further change for patients in Classes 1 and 2 and a stable course for patients in Class 3. In a repeated measures mixed model controlling for potential confounders, class was a significant predictor of functional status. The effect of baseline depression score, cognitive status, self-perceived health and sex on mobility/IADL score differed by class. CONCLUSIONS: These findings show systematic variability in functional status over time among older patients with major depression, indicating that a single trajectory may not reflect the pattern for all patients. PMID- 19548212 TI - Reversed phase liquid chromatography trace analysis of pesticides in soil by on column sample pumping large volume injection and UV detection. AB - The idea of utilization of one hydraulic line of a common commercial HPLC pump for direct on-column sample pumping injection of large sample volumes, 20 mL, was further investigated with the aim to develop multicomponent pesticides trace residues HPLC method in gram soil samples. Target pesticides group involve asulam, atrazine, 2,4-D, PCA, propazine, simazine, 4-chloro-2-methylphenoxyacetic acid, 2-(4-chloro-2-tolyloxy) propionic acid, chlortoluron, metoxuron, epoxiconazole. The results proved the applicability of this approach in experiments with mixtures of analytes at low ng/mL levels. Analysis of 20 mL of soil leachates and extracts of fortified soil samples containing these pesticides at the 10-50 ng/g level (in dry soil) revealed good figures of merit, also in the presence of large excess of humics. LODs achieved by detection at 220 nm evaluated from calibration runs of spiked soil extracts by Hubaux et al. method ranged from 5-12 ng per injected volume. For 20 mL large volume injection it represents 0.25-0.6 ng/mL of diluted soil extract, or 2.5-6 ng/mL of crude extract, or 6-5 ng/g dry soil. Recoveries of pesticides at concentration levels approaching half of maximum allowable concentration of pesticides in soil (100 ng/g) ranged from 85 to 98% with acceptable reproducibility, except asulam and metoxuron. PMID- 19548213 TI - Dispersive liquid-liquid microextraction followed by gas chromatography-electron capture detection for determination of polychlorinated biphenyls in fish. AB - A new method of dispersive liquid-liquid microextraction (DLLME) combined with GC electron capture detection (GC-ECD) was proposed for the extraction and determination of four polychlorinated biphenyls (PCBs) congeners in fish samples. Acetone was used as extraction solvent for the extraction of PCBs from fish samples. The target analytes in the acetone solvent were rapidly transferred to chlorobenzene, which was used as extraction solvent in DLLME procedures. Under the optimum conditions, linearity was obtained in the concentration range from 1.25 to 1250 microg/kg for PCB 52, and 0.25 to 250 microg/kg for PCB 101, 138 and 153. Coefficients of correlation (r2) ranged from 0.9993 to 0.9999. The repeatability was tested by spiking fish samples at 10 microg/kg PCBs, and RSD% (n = 8) varied between 2.2 and 8.4%. The LODs were between 0.12 and 0.35 microg/kg. The enrichment factors of PCBs were from 87 to 123. The relative recoveries of the four PCB congeners for the perch, pomfret and yellow-fin tuna at spiking levels of 10, 20 and 50 microg/kg were in the range of 81.20-100.6%, 85.00-102.7% and 87.80-108.4%, respectively. The results demonstrated that DLLME combined with GC-ECD was a simple, rapid, and efficient technique for the extraction and determination of PCBs in fish samples. PMID- 19548214 TI - Determination of nonylphenol and nonylphenol ethoxylates in wastewater using MEKC. AB - Nonylphenol ethoxylates (NPEO(x)) are surfactants which are used worldwide and can be transformed in the environment by microorganisms to form nonylphenol (NP). Analysis of these compounds was carried out with micellar electrokinetic capillary chromatography (MEKC). Different parameters such as background electrolyte (BGE) solution, pH, type of surfactant, and sample stacking were optimized. The use of CHES (20 mM, pH 9.1) in combination with 50 mM sodium cholate as a surfactant as BGE solution, together with sample stacking using 50 mM NaCl in the sample and an injection time of 20 s, provided the best separation of the compounds studied. The method was applied to the determination of target analytes in two types of sludge water coming from two steps of a wastewater treatment plant. Liquid-liquid extraction was carried out using toluene as solvent, resulting in recoveries around 100% for all studied analytes. The presence of NPEO(x) was observed in the first step of the sludge water treatment, based on migration time and UV spectra. Identification was confirmed using tandem MS. LOQs of the studied compounds were in the range of 12.7 to 30.8 ng/mL, which is satisfactory for the analysis of real wastewater samples. PMID- 19548215 TI - Quantification of domoic acid in shellfish tissues by pressurized capillary electrochromatography. AB - A method was developed to quantify domoic acid (DA), the chemical responsible for amnesic shellfish poisoning (ASP), by pressurized CEC (pCEC). The effect of different experimental conditions on the separation of DA and matrix solutes, such as the content of ACN in mobile phase, pH and concentration of buffer, supplementary pressure and applied voltage, were investigated. Under the optimal conditions, the pCEC method separated DA from shellfish matrices within 6 min. By using supplementary pressure, bubble formation in the capillary column was completely suppressed. The method was repeatable, sufficient accurate and sensitive for rapid screening of DA in shell seafood. PMID- 19548216 TI - Preparation of functionalized magnetic nanoparticulate sorbents for rapid extraction of biphenolic pollutants from environmental samples. AB - A new solid-phase extraction coupled with magnetic carrier technology was developed for extraction of bisphenol A (BPA) and diethylstilbestrol (DES) from water samples. The SPE sorbents, functionalized magnetic nanoparticles (Fe3O4@SiO2/beta-CD, core/shell), were synthesized in a two-stage system. The material was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and a vibrating sample magnetometer. SPE extraction parameters, such as volume and pH of sample, adsorption time, and desorption conditions were optimized. Under selected conditions: 250 mL of water sample, 0.1 g of sorbents and elution with methanol (3 mL with 1% acetic acid), the extraction was completed in 25 min. SPE followed by HPLC was employed to determine BPA and DES in environmental samples. The developed method provided spiked recoveries of 80-105%, relative standard deviations of less than 7%, and LOD of BPA (20.0 ng/L) and DES (23.0 ng/L), respectively. The proposed method offered easy preparation of sorbents, rapid analysis, high enrichment yields, and reliable quantitative assay. PMID- 19548217 TI - A capillary zone electrophoresis for determination of thiolic peptides in biological samples. AB - A new method to improve the analyses of thiolic peptides (cysteine, gammaGlu-Cys, glutathione, phytochelatins and desglycyl-phytochelatins) derivatized with monobromobimane (mBrB) in complex biological samples by CZE is described. The method involves a SPE using Sep-Pak Light C18 Cartridges after derivatization and a later CZE analysis. Elution of mBrB-thiols was achieved with 10 mM HCl + 70% methanol v/v in deionised water. Electrophoretic parameters, such as BGE pH and concentration, different organic additives (methanol and trifluoroethanol), applied voltage and capillary length were studied in order to establish suitable analytical conditions. Optimum separation of the mBrB-thiolic peptides was obtained with 100 mM sodium borate buffer at pH 7.60. The electrophoretic conditions were +15 kV, capillary length of 90 cm from inlet to detector (98 cm total length, 50 microm ID), samples were loaded into the capillary by hydrodynamic injection (50 mbar, 20 s) and detection was performed at 390 nm. The improved method showed good reproducibility, linearity and sensitivity. The LODs and LOQs estimated using a standard of GSH were 1.41 and 4.69 microM respectively. PMID- 19548218 TI - Modeling and prediction of retention behavior of histidine-containing peptides in immobilized metal-affinity chromatography. AB - Two kinds of structural characterization method as local descriptors and global descriptors were used to parameterize peptide structures, and several quantitative structure-retention relationship models were then constructed using partial least square (PLS), least-squares support vector machine (LS-SVM) and Gaussian process (GP) coupled with genetic algorithm-variable selection. These models were validated rigorously and investigated systematically by Tropsha et al. criteria, Monte Carlo cross-validation and one-way analysis of variance. Results show that regression models constructed using nonlinear approaches such as LS-SVM and GP are more robust and predictable than those by linear PLS method. By including linear and nonlinear terms in the covariance function, the GP is capable of handling both linear and nonlinear-mixed relationship, and thus presents a better performance than LS-SVM. Investigation of the optimal GP model revealed that diversified properties contribute to the retention behavior of peptides in immobilized metal-affinity chromatography. Particularly, coordination interaction, electrostatic factor, sovlation effect and hydrogen bonding are correlated significantly with the peptide retention ability. PMID- 19548221 TI - Nocturnal sleep duration and cognitive impairment in a population-based study of older adults. AB - OBJECTIVE: We aimed to investigate the association between nocturnal sleep duration, changes in nocturnal sleep duration and cognitive impairment in older adults. METHODS: 4010 participants of a population-based cohort study provided information on nocturnal sleep duration at baseline (1991-1995) and at follow-up (2002/2003). 792 follow-up participants aged 70+ by 2006 participated in telephone-based cognitive assessments. Several cognitive tests were used including the telephone interview for cognitive status (TICS). Cognitive impairment was defined as <31 points on the TICS (13.0%) and as below this percentile on the other tests. Based on individual tests, a verbal memory score and a total score were constructed. Multivariable prevalence ratios (PRs) of cognitive impairment and 95% confidence intervals (95%CIs) were computed using Poisson regression. Analyses were restricted to those free of depression in 2002/2003 (n = 695). RESULTS: Sleeping or=9 h was positively, although imprecisely, associated with impairment of verbal memory (PR = 1.7, 95%CI = 1.0, 3.0), and less pronounced with the other cognitive measures. An increase in sleep duration from 7-8 h in 1992-1995 to >or=9 h 8.5 years later (versus sleeping 7-8 h at both time points) was associated with an increased prevalence of cognitive impairment according to the TICS (PR = 2.1, 95% = 1.0, 4.5) and the verbal memory score (PR = 2.0, 95%CI = 1.0, 3.8). CONCLUSIONS: Increases in sleep duration are associated with cognitive impairment. A biological explanation for this association is currently lacking. Increases in sleep duration could be a marker of cognitive deficits. PMID- 19548222 TI - Changes in utilisation of anticholinergic drugs after initiation of cholinesterase inhibitors. AB - PURPOSE: Cholinesterase inhibitors (CEIs) have been subsidised in Australia since February 2001 for cognitive decline associated with mild to moderate Alzheimer disease. The number of people with Alzheimer disease is expected to increase, with a continuing increase in the number of people receiving CEI's. Many anticholinergic drugs (ACDs) are also prescribed to people receiving CEIs and concerns about the impact of the interaction have been raised. The aim of this study was to describe co-prescribing of a group of important ACDs in patients initiating treatment with CEIs in Australia. METHODS: Pharmacy claim data for Australia (Pharmaceutical Benefits Scheme) was examined for the period 1 April to 30 June 2006. All selected prescriptions supplied for patients receiving their first supply of any CEIs (initiators) were extracted for 14 weeks prior to and post the first date of supply. The numbers of initiating people co-administering CEIs and ACDs was examined. RESULTS: 5797 persons received their first prescription for CEIs between 1 April and 30 June 2006. Thirty-two per cent of these also received prescriptions for at least one ACD. There was a statistically significant increase in the number of initiators receiving an ACD. The significant increase was in patients receiving atypical antipsychotics. There was a trend towards an increase in patients receiving oxybutynin. CONCLUSIONS: Extent of co-administration of ACDs and CEIs is similar to other international studies however the most significant increase is seen in patients receiving atypical antipsychotics. The implications of adding atypical antipsychotics are potential for worsening disease, increasing adverse effects and increased health resource utilisation in this vulnerable group. PMID- 19548223 TI - Hospital pharmacists' activities and attitudes regarding the Thai safety monitoring program for new drugs. AB - PURPOSE: To determine the knowledge, attitudes and practices of Thai hospital pharmacists concerning the safety monitoring programme and reporting of adverse drug reactions to the Thai Food and Drug Administration. METHODS: Postal questionnaires distributed to 414 Thai hospitals, including all larger hospitals and a proportion of small community hospitals, for completion by the pharmacist responsible for ADR reporting to the Thai FDA. RESULTS: The response rate was 63.0% (n = 262) with most respondents (190; 72.8%) being directly responsible for ADR reporting. Most hospitals (251; 96.5%) designated at least one pharmacist as responsible for ADR reporting, but only 164 (62.8%) respondents were aware of the SMP. Only 15 respondents out of the 56 working in hospitals using new drugs had established reporting via the SMP. Uncertainty in ascribing causality, lack of co operation between health professionals and lack of staff were identified as barriers to reporting. Most respondents agreed that enhanced co-operation and improvements needed in hospital systems were required. Less than half agreed that the reporting system was simple and the majority agreed that changes were required to the SMP reporting mechanisms. CONCLUSIONS: Thai hospital pharmacists have positive attitudes towards reporting of ADRs, but not all are aware of the SMP for new drugs. Few hospitals use new drugs and even less are involved in active reporting of suspected ADRs via the SMP. To increase reporting rates, potential difficulties with the organization of reporting via the SMP and the potential role of hospital pharmacists in ADR identification should be investigated further. PMID- 19548224 TI - Multiple steroid courses result in tumour shrinkage in congenital pulmonary airway malformation (congenital cystic adenomatoid malformation). PMID- 19548225 TI - Prenatal diagnosis of epidermolytic palmoplantar keratoderma caused by c.T470C (p.M157T) of the keratin 9 gene in a Chinese kindred. PMID- 19548227 TI - Infectious entertainment. PMID- 19548229 TI - Involvement of cellular proteins in Junin arenavirus entry. AB - Junin arenavirus (JUNV) entry is dependent on clathrin-mediated pathways and it relies on the integrity of the actin cytoskeleton as well as the dynamics of microtubules. To determine the method of entry used by this human pathogen, we have demonstrated that in Vero cells JUNV is trafficked via the cellular dynamin 2 (dyn2) endocytic pathway and it is dependent on the Eps15 GTPase. In addition, we have shown that the virus travels through Rab5-mediated early and Rab7 mediated late endosomes in its pH-dependent entry. Altogether, this study gives further inside into the endocytic pathway utilized by the arenavirus JUNV. PMID- 19548233 TI - Retrospective: Birth of the cool - imaging and microbiology from Ibn al-Haytham to Jean Comandon. PMID- 19548231 TI - Dual color localization microscopy of cellular nanostructures. AB - The dual color localization microscopy (2CLM) presented here is based on the principles of spectral precision distance microscopy (SPDM) with conventional autofluorescent proteins under special physical conditions. This technique allows us to measure the spatial distribution of single fluorescently labeled molecules in entire cells with an effective optical resolution comparable to macromolecular dimensions. Here, we describe the application of the 2CLM approach to the simultaneous nanoimaging of cellular structures using two fluorochrome types distinguished by different fluorescence emission wavelengths. The capabilities of 2CLM for studying the spatial organization of the genome in the mammalian cell nucleus are demonstrated for the relative distributions of two chromosomal proteins labeled with autofluorescent GFP and mRFP1 domains. The 2CLM images revealed quantitative information on their spatial relationships down to length scales of 30 nm. PMID- 19548236 TI - Tracking algorithms chase down pathogens. AB - Understanding subcellular dynamic processes governing pathogenic mechanisms is a necessary step towards the development of new drugs and strategies against infectious diseases. Subcellular pathogenic mechanisms, such as viral invasion processes involve highly dynamic nanometric-scale objects and rapid molecular interactions that require the study of individual particle paths. Single-particle tracking methods allow visualizing and characterizing the dynamics of biological objects, and provide a straightforward and accurate means to understand subcellular processes. This review describes a number of particle-tracking methods in time-lapse microscopy sequences and provides examples of using such techniques to investigate mechanisms of host-pathogen interactions. PMID- 19548237 TI - PALM and STORM: what hides beyond the Rayleigh limit? AB - Super-resolution imaging allows the imaging of fluorescently labeled probes at a resolution of just tens of nanometers, surpassing classic light microscopy by at least one order of magnitude. Recent advances such as the development of photo switchable fluorophores, high-sensitivity microscopes and single particle localization algorithms make super-resolution imaging rapidly accessible to the wider life sciences research community. As we take our first steps in deciphering the roles and behaviors of individual molecules inside their living cellular environment, a new world of research opportunities beckons. Here we discuss some of the latest developments achieved with these techniques and emerging areas where super-resolution will give fundamental new "eye" sight to cell biology. PMID- 19548239 TI - Editorial: Imaging host-pathogen interactions. PMID- 19548242 TI - Meeting report: Imaging host-pathogen interactions. PMID- 19548243 TI - Careers in the twilight between bugs and cells. PMID- 19548244 TI - Effector T-cell differentiation during viral and bacterial infections: Role of direct IL-12 signals for cell fate decision of CD8(+) T cells. AB - To study the role of IL-12 as a third signal for T-cell activation and differentiation in vivo, direct IL-12 signaling to CD8(+) T cells was analyzed in bacterial and viral infections using the P14 T-cell adoptive transfer model with CD8(+) T cells that lack the IL-12 receptor. Results indicate that CD8(+) T cells deficient in IL-12 signaling were impaired in clonal expansion after Listeria monocytogenes infection but not after infection with lymphocytic choriomeningitis virus, vaccinia virus or vesicular stomatitis virus. Although limited in clonal expansion after Listeria infection, CD8(+) T cells deficient in IL-12 signaling exhibited normal degranulation activity, cytolytic functions, and secretion of IFN-gamma and TNF-alpha. However, CD8(+) T cells lacking IL-12 signaling failed to up-regulate KLRG1 and to down-regulate CD127 in the context of Listeria but not viral infections. Thus, direct IL-12 signaling to CD8(+) T cells determines the cell fate decision between short-lived effector cells and memory precursor effector cells, which is dependent on pathogen-induced local cytokine milieu. PMID- 19548246 TI - Perspective: Hidden treasures from the archives. PMID- 19548245 TI - Effect of Terminalia chebula aqueous extract on oxidative stress and antioxidant status in the liver and kidney of young and aged rats. AB - We evaluated the preventive effects of Terminalia chebula (T. chebula) aqueous extract on oxidative and antioxidative status in liver and kidney of aged rats compared to young albino rats. The concentrations of malondialdehyde (MDA), lipofuscin (LF), protein carbonyls (PCO), activities of xantione oxidase (XO), manganese-superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), and glucose-6 phosphate dehydrogenase (G6PDH), levels of glutathione (GSH), vitamin C and vitamin E were used as biomarkers. In the liver and kidney of aged animals, enhanced oxidative stress was accompanied by compromised antioxidant defences. Administration of aqueous extract of T. cheubla effectively modulated oxidative stress and enhanced antioxidant status in the liver and kidney of aged rats. The results of the present study demonstrate that aqueous extract of T. cheubla inhibits the development of age-induced damages by protecting against oxidative stress. PMID- 19548247 TI - YouTube: Increasing your bug views. PMID- 19548248 TI - Hypomorphic mutation of ZAP70 in human results in a late onset immunodeficiency and no autoimmunity. AB - Complete lack of function of the tyrosine kinase ZAP70 in humans results in a severe immunodeficiency, characterized by a lack of mature CD8(+) T cells and non functional CD4(+) T cells. We report herein an immunodeficiency with an inherited hypomorphic mutation of ZAP70 due to a single G-to-A substitution in a non-coding intron. This mutation introduces a new acceptor splice site and allows low levels of normal alternative splicing and of WT ZAP70 expression. This partial deficiency results in a compromised TCR signaling that was totally restored by increased expression of ZAP70, demonstrating that defective activation of the patient T cells was indeed caused by the low level of ZAP70 expression. This partial ZAP70 deficiency was associated with an attenuated clinical and immunological phenotype as compared with complete ZAP70 deficiency. CD4(+) helper T-cell populations including, follicular helper T cells, Th1, Th17 and Treg were detected in the blood. Finally, the patient had no manifestation of autoimmunity suggesting that the T-cell tolerogenic functions were not compromised, in contrast to what has been observed in mice carrying hypomorphic mutations of Zap70. This report extends the phenotype spectrum of ZAP70 deficiency with a residual function of ZAP70. PMID- 19548249 TI - A systematic review of impulsivity in eating disorders. AB - The purpose of this paper is to conduct a systematic review of the current literature that examines impulsivity in individuals with eating disorders (ED). Studies were obtained from Embase, Pubmed and Psycinfo, and were included if they assessed impulsivity in individuals over 18 years of age with an ED diagnosis and published in the last 10 years. The methodological quality of the studies was rated. Twelve studies were included in this review, with methodological quality varying across studies. Findings suggest that impulsivity is best assessed multi modally, with a combination of self-report, behavioural and physiological measures. In general, impulsivity was found to differentiate individuals with EDs from controls, as well as across diagnostic subtypes. The current findings have important clinical implications for our understanding and treatment of both impulsivity and eating disorders. PMID- 19548250 TI - User satisfaction with services in a randomised controlled trial of adolescent anorexia nervosa. AB - BACKGROUND: User satisfaction is a neglected outcome in adolescent anorexia nervosa especially since the relative effectiveness of different treatments is unclear. It may also affect clinical outcome. AIMS: To assess young person's and parents' satisfaction with CAMHS outpatient, specialist outpatient and inpatient treatment received in a large randomised controlled trial. METHOD: Quantitative and qualitative analysis of questionnaire data from 215 young people and their parents followed by focus groups to further explore emerging themes. RESULTS: High levels of satisfaction were reported, more amongst parents than young people and with specialist services. Both young people and carers strongly valued clinical relationships that involved being listened to and understood. They valued the expertise of specialist rather than generic CAMHS services. There were polarised views on the influence of other young people in inpatient units. Parents in particular valued support for themselves, both from professionals and other parents and felt this, and sibling support was lacking. CONCLUSIONS: All comprehensive CAMH services are able to provide the good generic psychotherapeutic skills that parents and young people value so highly. However, generic CAMHS struggle to provide the demanded level of expertise and more specialised individual and family therapeutic interventions. PMID- 19548252 TI - A theoretical study of topomerization of imine systems: inversion, rotation or mixed mechanisms? AB - The different mechanisms, rotation, inversion, or intermediate mechanism, by which occur the topomerization of imine systems R(2) C=N-X have been studied by applying ab initio, B3LYP, and MP2 methods. The effect of a wide variety of substituents R and X on the isomerization pathway have been examined by computing fully optimized structures of the ground and transition states (136 isomers belonging to different imine families were studied and more than 300 transition structures were determined at various levels of theory). Energy barriers have been also obtained and it was found that the groups R and X have a strong influence on the type of mechanism involved and the activation energies. Thus, and depending on the type of substituents, transition state structures related to the following kinds of processes were found: pure inversion, intermediate mechanisms, rotation, and enhanced rotation (hyper-rotation). In turn, the corresponding activation energies range between very low (<10 kcal/mol) and extremely high (> 70 kcal/mol) values. A simple index that allows us to quantify the percentage of inversion or rotation mechanism is proposed. PMID- 19548253 TI - A processing method enabling the use of peak height for accurate and precise proton NMR quantitation. AB - In NMR, peak area quantitation is the most common method used because the area under a peak or peak group is proportional to the number of nuclei at those frequencies. Peak height quantitation has not enjoyed as much utility because of poor precision and linearity as a result of inconsistent shapes and peak widths (measured at half height). By using a post-acquisition processing method employing a Gaussian or line-broadening (exponential decay) apodization (i.e. weighting function) to normalize the shape and width of the internal standard (ISTD) peak, the heights of an analyte calibration spectrum can be compared to the analyte peaks in a sample spectrum resulting in accurate and precise quantitative results. Peak height results compared favorably with 'clean' peak area results for several hundred illicit samples of methamphetamine HCl, cocaine HCl, and heroin HCl, of varying composition and purity. Using peak height and peak area results together can enhance the confidence in the reported purity value; a major advantage in high throughput, automated quantitative analyses. PMID- 19548254 TI - Oligonol, a new lychee fruit-derived low-molecular form of polyphenol, enhances lipolysis in primary rat adipocytes through activation of the ERK1/2 pathway. AB - The effect of Oligonol, a phenolic product from lychee fruit polyphenol (LFP) containing catechin-type monomers and lower oligomers of proanthocyanidin, on lipolysis in primary adipocytes was investigated in order to examine the possible mechanism underlying the regulation of in vivo metabolism in fat. Oligonol significantly increased lipolysis, which was accompanied by both activation of extracellular signaling-related kinase 1/2 (ERK1/2) and down-regulation of perilipin protein expression, without an increase in intracellular cAMP production. The increase in lipolysis with Oligonol was prevented completely by pretreatment with either PD98059 or U0126, selective ERK1/2 inhibitors, which also prevented the reduction in the expression of perilipin protein. Tumor necrosis factor-alpha also down-regulated the expression of perilipin protein. However, there was no significant alteration in the expression of Galphai protein with Oligonol. These findings indicate that Oligonol enhances lipolysis in primary adipocytes, independent of cAMP production, but its effect is dependent on activation of the ERK1/2 pathway, leading to down-regulation of perilipin protein expression. PMID- 19548255 TI - CAG-repeat length and the age of onset in Huntington disease (HD): a review and validation study of statistical approaches. AB - CAG-repeat length in the gene for HD is inversely correlated with age of onset (AOO). A number of statistical models elucidating the relationship between CAG length and AOO have recently been published. In the present article, we review the published formulae, summarize essential differences in participant sources, statistical methodologies, and predictive results. We argue that unrepresentative sampling and failure to use appropriate survival analysis methodology may have substantially biased much of the literature. We also explain why the survival analysis perspective is necessary if any such model is to undergo prospective validation. We use prospective diagnostic data from the PREDICT-HD longitudinal study of CAG-expanded participants to test conditional predictions derived from two survival models of AOO of HD. A prior model of the relationship of CAG and AOO originally published by Langbehn et al. yields reasonably accurate predictions, while a similar model by Gutierrez and MacDonald substantially overestimates diagnosis risk for all but the highest risk participants in this sample. The Langbehn et al. model appears accurate enough to have substantial utility in various research contexts. We also emphasize remaining caveats, many of which are relevant for any direct application to genetic counseling. PMID- 19548256 TI - Association of MET with social and communication phenotypes in individuals with autism spectrum disorder. AB - Autism is a complex neurodevelopmental disorder diagnosed by impairments in social interaction, communication, and behavioral flexibility. Autism is highly heritable, but it is not known whether a genetic risk factor contributes to all three core domains of the disorder or autism results from the confluence of multiple genetic risk factors for each domain. We and others reported previously association of variants in the gene encoding the MET receptor tyrosine kinase in five independent samples. We further described enriched association of the MET promoter variant rs1858830 C allele in families with co-occurring autism and gastrointestinal conditions. To test the contribution of this functional MET promoter variant to the domains of autism, we analyzed its association with quantitative scores derived from three instruments used to diagnose and describe autism phenotypes: the Autism Diagnostic Interview-Revised (ADI-R), the Autism Diagnostic Observation Schedule (ADOS), and both the parent and the teacher report forms of the Social Responsiveness Scale (SRS). In 748 individuals from 367 families, the transmission of the MET C allele from parent to child was consistently associated with both social and communication phenotypes of autism. Stratification by gastrointestinal conditions revealed a similar pattern of association with both social and communication phenotypes in 242 individuals with autism from 118 families with co-occurring gastrointestinal conditions, but a lack of association with any domain in 181 individuals from 96 families with ASD and no co-occurring gastrointestinal condition. These data indicate that the MET C allele influences at least two of the three domains of the autism triad. PMID- 19548258 TI - Commentary: The federal 'Prenatally and Postnatally Diagnosed Conditions Awareness Act'. AB - The recently enacted federal law, the 'Prenatally and Postnatally Diagnosed Conditions Awareness Act' (United States Public Law 110-374) seeks to improve opportunities for parents and pregnant women to anticipate and understand the likely life course of children born with Down syndrome and other (unspecified) conditions. The law is in part a response to the continued growth of prenatal screening and testing. For example, the American College of Obstetricians and Gynecologists' Practice Bulletin 77 recommends that 'Screening and invasive diagnostic testing for aneuploidies be available to all women who present for prenatal care before 20 weeks of gestation regardless of maternal age.' Emerging technologies anticipate an era in which the scope of prenatal screening and testing will be much larger than it is today. Inevitably, more women will find themselves facing the hard question of whether to continue or end a pregnancy in which a fetus has been found to have a significant abnormality. While the new federal law is not likely to have a major impact on obstetric practice, it may be a harbinger of renewed wide-scale public debate concerning the ethics of prenatal screening. PMID- 19548257 TI - Screening of some Kenyan medicinal plants for antibacterial activity. AB - Eleven medicinal plants used by traditional healers in Machakos and Kitui District were screened, namely: Ajuga remota Benth, Aloe secundiflora Engl, Amaranthus hybridus L, Cassia didymobotrya Fes, Croton macrostachyus Del, Entada leptostachya Harms, Erythrina abyssinica DC, Harrisonia abyssinica Oliv, Schkuhria pinnata O. Ktze, Terminalia kilimandscharica Engl and Ziziphus abyssinica Hochst for potential antibacterial activity against four medically important bacterial strains, namely: Bacillus cereus ATCC 11778, Escherichia coli ATCC 25922, Micrococcus lutea ATCC 9341 and Pseudomonas aeruginosa ATCC 27853. The antibacterial activity of methanol extracts was determined as the minimum inhibitory concentration (MIC). The plant extracts were more active against Gram positive (G+) than Gram-negative (G-) bacteria. The positive controls were streptomycin and benzylpenicillin for G- and G+ bacteria, respectively, both had a significant MIC at <1 mg/mL. The most susceptible bacteria were B. cereus, followed by M. lutea, while the most resistant bacteria were Ps. aeruginosa, followed by E. coli. The present study supports the use of these plants by the herbalists in the management of bacterial ailments. H. abyssinica and T. kilimandscharica showed the best antibacterial activity; hence these plants can be further subjected to phytochemical and pharmacological evaluation. PMID- 19548259 TI - Prediction model of lymph node metastasis in superficial esophageal adenocarcinoma and squamous cell cancer including D2-40 immunostaining. AB - BACKGROUND: It was the aim of our study to establish a model for prediction of lymph node metastases in superficial esophageal cancer. METHODS: We analyzed the clinical and histopathological data of 50 consecutive patients with pT1 esophageal cancer who underwent oncological resection. Submucosal carcinomas (pT1b) were classified according to sm levels 1-3. D2-40 immunostaining was investigated using the ABC technique. In a first step, we performed univariate analysis (One-way ANOVA: Sigma restricted parameterization; test of SS whole vs. SS predicted) to test the predictive value of the following categorical parameters for lymph node status (positive/negative): sex, histologic tumor type, localization, surgical technique (transhiatal/transthoracic), grading, pT1 subclassification (pT1a, pT1b sm 1-3), pL-, pV-status, and D2-40 labeling. Simple regression was applied for the following continuous predictors: age and tumor size. All significant variables of univariate analysis were included in the multivariate analysis. For this purpose, we used the General Liner Models's analysis (forward stepwise). In a third step, the Kruskal-Wallis test with post hoc comparisons was intended to define the cut-off value of parameters tested. RESULTS: Only the following variables gained statistical significance in univariate analysis: sex, histological tumor type, grading, pT1 subclassification, lymphatic infiltration, microvascular infiltration, D2-40 immunostaining, and tumor size (P < 0.05). Variables reaching significance in multivariate analysis were tumor size (P = 0.017) and pV-status (P = 0.037). In the Kruskal-Wallis test with post hoc comparisons, the cut-off value of tumor size was 2 cm (model P = 0.002) and between the categories (P < 0.05). CONCLUSIONS: Lymph node positivity and lymphatic vessel infiltration did not linearly increase with sm tumor infiltration depth. The risk category of lymph node involvement in superficial esophageal cancer exists according to our prediction model on the basis of tumor size of >2 cm and microvascular infiltration. The hitherto common sm levels 1-3 classification of submucosal cancers appears to display a lesser impact than previously assumed with regard to prediction of potential lymph node metastases and consequently the indication for endoscopic or surgical therapy. PMID- 19548260 TI - A longitudinal follow-up study of autistic symptoms in children and adults with duplications of 15q11-13. AB - We completed a longitudinal follow-up of autistic symptoms in a cohort of children and young adults with duplications of chromosome 15q11-13. In our initial investigation of 29 individuals, tentative conclusions were drawn based on cross-sectional data suggesting that autistic symptoms increased with age, most specifically in the area of social interaction. We were able to re-assess 22 individuals from the original study an average of 7 years later using the same standardized autism screening measure. As predicted, autistic symptoms were found to increase for the younger children in the cohort but remained constant for the older participants. This global change in autistic symptoms among the younger children could not be ascribed to any particular autism subscale and reflects small but cumulatively significant increases across several domains. PMID- 19548261 TI - Cancer mortality in patients with schizophrenia: an 11-year prospective cohort study. AB - BACKGROUND: Schizophrenia has been associated with a rate of premature mortality that is 2 to 3 times higher than that in the general population. Although the role of cancer in this excess mortality remains unclear, previous incidence or mortality studies found contradictory results. METHODS: In 1993, a large prospective study was initiated in a cohort of 3470 patients with schizophrenia to examine cancer-related mortality and predictors. Standardized mortality ratios (SMRs) were calculated, adjusting for age and sex relative to a representative sample of the French general population. RESULTS: During the 11-year follow-up, 476 (14%) patients died; the mortality rate was thus nearly 4-fold higher than in the general population. Cancer was the second most frequent cause of mortality (n=74), with a global SMR of 1.5 (95% confidence interval [95% CI], 1.2-1.9). For all cancers, the SMRs were 1.4 (not significant) for men and 1.9 (95% CI, 1.4 2.8) for women. For men, lung cancer was the most frequent localization (n=23; 50%), with an SMR of 2.2 (95% CI, 1.6-3.3). For women, breast cancer was the most frequent localization (n=11; 39%), with an SMR of 2.8 (95% CI, 1.6-4.9). In comparison with patients who did not die of cancer, there were 2 significant baseline predictors of death by lung cancer in the final logistic regression model: duration of smoking and age>38 years. CONCLUSIONS: The results of the current study demonstrated an increased risk of mortality by cancer in patients with schizophrenia, especially for women from breast cancer and for men from lung cancer. PMID- 19548262 TI - Ewing sarcoma demonstrates racial disparities in incidence-related and sex related differences in outcome: an analysis of 1631 cases from the SEER database, 1973-2005. AB - BACKGROUND: Previous reports of Ewing sarcoma cohorts suggested that there is a difference in incidence according to racial origin. However, to the authors' knowledge, this finding has never been tested in a population-based database, and the impact of race on clinical outcome and the significance of known risk factors stratified to racial groups have not been reported. METHODS: Patients who had Ewing sarcoma diagnosed between 1973 and 2005 were identified in the Surveillance, Epidemiology, and End Results database. Patient demographic and clinical characteristics; incidence; year of diagnosis; tumor location, tumor size, and disease stage at diagnosis; treatment(s); cause of death; and survival were extracted. Kaplan-Meier, log-rank, and Cox regressions were used to analyze the significance of prognostic factors. RESULTS: Race-specific incidence indicated that Caucasians have the highest incidence (0.155), followed by Asians/Pacific Islanders (0.082), and African Americans (0.017). The difference in incidence between Caucasians and African Americans was 9-fold and significant (P<.001). The incidence of Ewing sarcoma increased over the past 3 decades among Caucasians (P<.05). Survival was not impacted by race. Local disease stage, primary tumor location in the appendicular skeleton, and tumor size or =60) with clinically significant compulsive hoarding. They were assessed using structured interviews, including the Mini International Neuropsychiatric Interview (MINI), Structured Clinical Interview for DSM-IV (SCID I), Yale-Brown Obsessive Compulsive Scale (YBOCS), and UCLA Hoarding Severity Scale (UHSS). Self-report Measures Included the Beck Anxiety Inventory (BAI), Beck Depression Inventory (BDI), Sheehan Disability Scale (SDS), and Savings Inventory-Revised (SI-R). Psychosocial and medical histories were also obtained. To determine age at onset, participants were asked to rate their hoarding symptoms and describe major life events that occurred during each decade of their lives. RESULTS: Results show that (1) onset of compulsive hoarding symptoms was initially reported as being in mid-life but actually found to be in childhood or adolescence. No subjects reported late onset compulsive hoarding. (2) Compulsive hoarding severity increased with each decade of life. (3) Comorbid mood and anxiety disorders were common, but only 16% of patients met criteria for OCD if hoarding symptoms were not counted toward the diagnosis. (4) The vast majority of patients had never received treatment for hoarding. (5) Older adults with compulsive hoarding were usually socially impaired and living alone. CONCLUSIONS: Compulsive hoarding is a progressive and chronic condition that begins early in life. Left untreated, its severity increases with age. Compulsive hoarding should be considered a distinct clinical syndrome, separate from OCD. Unfortunately, compulsive hoarding is largely unrecognized and untreated in older adults. PMID- 19548274 TI - Stimulation of glucose uptake by triterpenoids from Weigela subsessilis. AB - Four ursane-type triterpenoids, corosolic acid (1), ilekudinol B (2), ursolic acid (3) and pomolic acid (4), were isolated from an EtOAc-soluble extract of the leaves of Weigela subsessilis. These bioactive compounds were evaluated for their glucose uptake activity and produced moderate to strong enhancement both in basal and insulin-stimulated L6 muscle cells. In particular, corosolic acid exhibited the most potent activity, increasing uptake by basal- and insulin-stimulated myotubes by 2.63- and 3.33-fold, respectively; ilekudinol B produced 1.6- and 2.9 fold, ursolic acid produced 1.84- and 2.64-fold, and pomolic acid produced 1.6- and 2.8-fold increases. No cytotoxicities were observed for corosolic acid, ursolic acid and ilekudinol B in myoblasts, while pomolic acid at doses of 25 and 50 microm reduced cell viability by 19% and 21.8% upon 24 h treatment and by 48.6% and 54.1% upon 48 h treatment, respectively. These results suggest that ursane-type triterpenoids from W. subsessilis might enhance glucose uptake by acting as insulin mimics and as insulin sensitizers and that they could be useful as nontoxic diabetes treatment agents. PMID- 19548273 TI - The ADAS-cog and clinically meaningful change in the VISTA clinical trial of galantamine for Alzheimer's disease. AB - BACKGROUND: A minimum 4-point change at 6 months on the Alzheimer's disease assessment scale-cognitive subscale (ADAS-cog) is deemed clinically important, but this cut-point has been little studied in relation to clinical meaningfulness. In an investigator-initiated, clinical trial of galantamine, we investigated the extent to which a 4-point change classifies goal attainment by individual patients. METHODS: Secondary analysis of the video imaging synthesis of treating Alzheimer's disease (VISTA) study: a 4-month, multi-centre, parallel group, double-blind, placebo-controlled, trial of galantamine in 130 mild moderate Alzheimer's disease patients (4-month open-label follow-up). ADAS-cog responses at 6 months were compared with outcomes on three clinical measures: clinician's interview based impression of change-plus caregiver input (CIBIC+), patient/carer-goal attainment scaling (PGAS) and clinician-GAS (CGAS). RESULTS: Thirty-seven of 99 patients improved by > or = 4 points on the ADAS-cog at 6 months, and 16/99 showed > or = 4-point worsening. ADAS-cog change scores correlated notionally to modestly with changes on the CGAS (r = -0.31), the PGAS (r = -0.29) and the CIBIC+ (r = 0.31). As a group, patients with ADAS-cog improvement were significantly more likely to improve on the clinical measures; those who worsened showed non-significant clinical decline. Individually, about half were misclassified in relation to each clinical measure; often when the ADAS Cog detected 'no change', clinically meaningful effects could be detected. Even so, no ADAS-Cog cut-point optimally classified patients' clinical responses. CONCLUSION: A 4-point ADAS-cog change at 6 months is clinically meaningful for groups. Substantial individual misclassification between the ADAS-cog and clinical measures suggests no inherent meaning to a 4-point ADAS-cog change for a given patient. PMID- 19548275 TI - Analgesic and antiinflammatory activity of Morinda citrifolia L. (Noni) fruit. AB - M. citrifolia is a tropical plant with a long tradition of medicinal use in Polynesia and tropical parts of eastern Asia and Australia. One of its favorite uses is the treatment of painful inflammatory conditions, such as arthritis. The analgesic activity of Noni fruit puree on mice was investigated using the hot plate test. A 10% solution of freeze concentrated Noni fruit puree in the drinking water of mice reduced the pain sensitivity comparably to the central analgesic drug tramadol. This effect was only partly reversed by the application of the morphine antagonist naloxone. An alcohol extract of noni fruit puree also caused an inhibition of MMP-9 release from human monocytes after stimulation with LPS. This effect was comparable to hydrocortisone (10(-5) m). The findings suggest that preparations of noni fruits are effective in decreasing pain and joint destruction caused by arthritis. PMID- 19548276 TI - Spatiotemporal control of apical and basal living subcellular chemical environments through vertical phase separation. AB - Molecular distribution within living cells is organized through multiscaled compartmentalization that enables specialized processes to occur with high efficiency. The ability to control the chemical environment at a subcellular level is limited due to deficient positional control over the aqueous stimulant. Here, a multilayered microfluidic system built from polydimethylsiloxane to separate chemical stimulants over single living cells vertically through aqueous phase separation under laminar flow is demonstrated. Cells are cultured on top of single micrometer-scale channels inside a larger channel, allowing labeling of the apical domain of single cells through the main channel with simultaneous and distinct labeling of the basal domain via the lower microchannels. The system is transparent, which allows the use of optical microscopy to investigate the spatiotemporal response of labeled components. By employing this technique, the examination of localized subcellular domain responses in polarization, lipid bilayer mobility, and apical-to-basal signal transduction can be explored. PMID- 19548277 TI - The interplay of crystallization kinetics and morphology in nanostructured W/Mo oxide formation: an in situ diffraction study. PMID- 19548278 TI - Orientation-dependent optical-polarization properties of single quantum dots in nanowires. PMID- 19548279 TI - Controlled assembly of highly Raman-enhancing silver nanocap arrays templated by porous anodic alumina membranes. AB - A convenient nanoscale technique is reported for the fabrication of highly ordered hemispherical silver nanocap arrays templated by porous anodic alumina (PAA) membranes as robust and cost-efficient surface-enhanced Raman scattering (SERS) substrates. This geometry produces a high Raman signal due to its periodic hexagonal arrangements and control of the gap between the nanostructures in the sub-10-nm regime. The surface structure can be tuned further to optimize the enhancement factor according to optional PAA fabrication and silver deposition parameters. Finite-difference time-domain calculations indicate that the structure may possess excellent SERS characteristics due to the high density and abundance of hot spots. PMID- 19548280 TI - Cytoprotective role of astaxanthin against glycated protein/iron chelate-induced toxicity in human umbilical vein endothelial cells. AB - Astaxanthin (ASX), a red carotenoid pigment with no pro-vitamin A activity, is a biological antioxidant that occurs naturally in a wide variety of plants, algae and seafoods. This study investigated whether ASX could inhibit glycated protein/iron chelate-induced toxicity in human umbilical-vein endothelial cells (HUVEC) by interfering with ROS generation in these cells. Glycated fetal bovine serum (GFBS) was prepared by incubating fetal bovine serum (FBS) with high concentration glucose. Stimulation of cultured HUVECs with 50 mm 1 mL of GFBS significantly enhanced lipid peroxidation and decreased antioxidant enzyme activities and levels of phase II enzymes. However, preincubation of the cultures with ASX resulted in a marked decrease in the level of lipid peroxide (LPO) and an increase in the levels of antioxidant enzymes in an ASX concentration dependent manner. These results demonstrate that ASX could inhibit LPO formation and enhance the antioxidant enzyme status in GFBS/iron chelate-exposed endothelial cells by suppressing ROS generation, thereby limiting the effects of the AGE-RAGE interaction. The results indicate that ASX could have a beneficial role against glycated protein/iron chelate-induced toxicity by preventing lipid and protein oxidation and increasing the activity of antioxidant enzymes. PMID- 19548281 TI - Enhanced solar-cell efficiency in bulk-heterojunction polymer systems obtained by nanoimprinting with commercially available AAO membrane filters. PMID- 19548282 TI - (-)-Epigallocatechin-3-gallate reduces experimental colon injury in rats by regulating macrophage and mast cell. AB - The ameliorative effect of (-)-epigallocatechin-3-gallate (EGCG) on inflammatory bowel disease (IBD) induced by ethanol 2,4,6-trinitrobenzene sulfonic acid (TNBS) was studied in 7-week-old male rats. Intestinal lesions were measured as an increase in myeloperoxidase (MPO) activity in mucosa. The supplementation of EGCG significantly inhibited MPO activity and histamine levels in the distal colon mucosa. The EGCG inhibited macrophage chemotaxis toward N-formyl-L-methionyl-L leucyl-L-phenylalanine in a concentration-dependent manner. These observations confirmed that EGCG can ameliorate acute experimental colitis by the suppression of mast cells and macrophage activities. PMID- 19548283 TI - Hesperidin suppressed proliferations of both human breast cancer and androgen dependent prostate cancer cells. AB - Hesperidin, a flavonoid derived from citrus fruits, has been reported to show various biological effects including anticancer activity. This study investigated whether hesperidin affected the proliferation of MCF-7 human breast cancer cells transfected with green fluorescent protein (GFP)/alpha-tubulin (MCF-7-GFP-Tubulin cells), androgen-independent PC-3 and DU-145 prostate cancer cells, and androgen dependent LNCaP prostate cancer cells. The results were as follows. (1) Hesperidin inhibited the proliferation of MCF-7-GFP-Tubulin cells, probably not through an antimitotic mechanism. (2) Hesperidin also inhibited both basal and testosterone-induced proliferation of LNCaP cells. (3) However, hesperidin did not significantly affect the cell proliferation of two hormone-independent prostate cancer cells, PC-3 and DU-145. It is concluded that hesperidin can inhibit the proliferation of breast cancer cells through mechanisms other than antimitosis and it is suggested that hesperidin be further investigated for the possible interaction with androgenic receptors and involvement in signaling pathway after receptor binding in prostate cancer cells through future research. PMID- 19548284 TI - The beta-carboline alkaloid harmine inhibits BCRP and can reverse resistance to the anticancer drugs mitoxantrone and camptothecin in breast cancer cells. AB - Multidrug resistance (MDR), mediated by highly expressed ABC transporters, is one of the most important mechanisms in tumor cells. Breast cancer resistance protein (BCRP) is a member of the ABC transporter family. This transporter expels different kinds of lipophilic anticancer drugs, which have diffused into the cells. In this study, 96-well plate based assays and flow cytometry analysis were employed to screen natural products for BCRP inhibition. The beta-carboline alkaloid harmine inhibited BCRP in a BCRP overexpressing breast cancer cell line (MDA-MB-231). Harmine reduced resistance to the anticancer drugs mitoxantrone and camptothecin mediated by BCRP and might be an interesting new reversal agent. Harmine did not inhibit P-glycoprotein (P-gp) mediated drug efflux. PMID- 19548285 TI - Household epidemics: modelling effects of early stage vaccination. AB - A Markovian susceptible --> infectious --> removed (SIR) epidemic model is considered in a community partitioned into households. A vaccination strategy, which is implemented during the early stages of the disease following the detection of infected individuals is proposed. In this strategy, the detection occurs while an individual is infectious and other susceptible household members are vaccinated without further delay. Expressions are derived for the influence on the reproduction numbers of this vaccination strategy for equal and unequal household sizes. We fit previously estimated parameters from influenza and use household distributions for Sweden and Tanzania census data. The results show that the reproduction number is much higher in Tanzania (6 compared with 2) due to larger households, and that infected individuals have to be detected (and household members vaccinated) after on average 5 days in Sweden and after 3.3 days in Tanzania, a much smaller difference. PMID- 19548286 TI - Thylakoids suppress appetite by increasing cholecystokinin resulting in lower food intake and body weight in high-fat fed mice. AB - Thylakoids are membranes isolated from plant chloroplasts which have previously been shown to inhibit pancreatic lipase/colipase catalysed hydrolysis of fat in vitro and induce short-term satiety in vivo. The purpose of the present study was to examine if dietary supplementation of thylakoids could affect food intake and body weight during long-term feeding in mice. Female apolipoprotein E-deficient mice were fed a high-fat diet containing 41% of fat by energy with and without thylakoids for 100 days. Mice fed the thylakoid-enriched diet had suppressed food intake, body weight gain and body fat compared with the high-fat fed control mice. Reduced serum glucose, serum triglyceride and serum free fatty acid levels were found in the thylakoid-treated animals. The satiety hormone cholecystokinin was elevated, suggesting this hormone mediates satiety. Leptin levels were reduced, reflecting a decreased fat mass. There was no sign of desensitization in the animals treated with thylakoids. The results suggest that thylakoids are useful to suppress appetite and body weight gain when supplemented to a high-fat food during long-term feeding. PMID- 19548287 TI - Protective effects of a flavonoid-rich extract of Hypericum perforatum L. against hydrogen peroxide-induced apoptosis in PC12 cells. AB - Hypericum perforatum L. has been used traditionally as an antidepressant for the treatment of mild to moderate depression. In a previous study, a flavonoid-rich extract of Hypericum perforatum L. (FEHP) was prepared and its antioxidant activity was determined by a series of models in vitro. In the present study, the protective effects of FEHP against hydrogen peroxide-induced apoptosis in rat pheochromocytoma line PC12 cells were investigated by MTT assay, lactate dehydrogenase (LDH) release assay, flow cytometry analysis and DNA fragmentation assay. Following a 4 h exposure of PC12 cells to H2O2, a significant decrease in the cell viability and increased levels of LDH release were observed. However, pretreatment of PC12 cells with FEHP prior to H2O2 exposure elevated the cell viability, decreased the levels of LDH release and decreased the occurrence of apoptotic cells. Also, the intensity of H2O2-induced DNA laddering was inhibited in a dose-dependent fashion by a DNA fragmentation assay. These results suggested that FEHP possessed protective effects against H2O2-induced apoptosis in PC12 cells and FEHP might be useful in the treatment of oxidative stress-related neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. PMID- 19548288 TI - Protective effect of oleanolic acid on gentamicin induced nephrotoxicity in rats. AB - Oleanolic acid is a molecule of current therapeutic interest. In the present study, oleanolic acid isolated from the cuticular epithelium of Viscum articulatum Burm. f. (Viscaceae) was investigated for its protective effects on gentamicin-induced renal damage in rats. Nephrotoxicity was induced in rats by intraperitoneal injection of gentamicin at a dose of 100 mg/kg/day for 8 days. The effect of Oleanolic acid administered orally at doses 40, 60 and 80 mg/kg/day was assessed biochemically by determination of albumin, urea and creatinine in serum and urine samples and also through histopathological examination of the kidneys. Oleanolic acid protected the rat kidneys from gentamicin-induced nephrotoxicity as evident from a decrease in the serum and urine levels of creatinine, albumin and urea. Oleanolic acid also protected the rat kidneys from histological alterations induced by gentamicin and also improved the glomerular filtration rate. Compared with an earlier report on intraperitoneal administration of oleanolic acid in paracetamol-induced nephrotoxicity in rats, the data show that orally administered oleanolic acid also exerted a nephroprotective effect even in the case of a nephrotoxicant such as gentamicin, which directly deteriorates the kidney function without prior metabolism. PMID- 19548289 TI - Stewartia koreana extract stimulates proliferation and migration of human endothelial cells and induces neovasculization in vivo. AB - Angiogenesis, the growth of new blood vessels from preexisting vasculature, plays an important role in physiological and pathological processes such as embryonic development and wound healing. This study investigated the effects of methanol extracts of Stewartia koreana leaves (SKE) on angiogenesis. Stewartia koreana significantly promoted the proliferation and migration of human umbilical vein endothelial cells in a dose-dependent manner. The SKE induced endothelial cell proliferation in the range of 50 microg/mL without cytotoxicity. Treatment of HUVECs resulted in the activation of the mitogen-activated protein kinases that was correlated with endothelial cell proliferation and migration. SKE also stimulated angiogenesis in a chick chorioallantoic membrane assay, demonstrating promotion of new blood vessel formation in vivo. Local administration of SKE onto skin punched wounds resulted in increased von Willebrand Factor antigen, indicating that it stimulated neovasculization in the wound region. The results suggest that Stewartia koreana extracts may potentially be useful for the development of agents to accelerate vascular wound healing or to promote the growth of collateral blood vessels in ischemic tissues. PMID- 19548290 TI - A systematic review on the sambuci fructus effect and efficacy profiles. AB - The berries of European elder are used in traditional German medicine for various complaints. Due to insufficient research data, elderberry fruit was not monographed by the German Commission E at the end of the last century. A comprehensive review of the literature was conducted to summarize the pharmacological and clinical effects of elderberry fruit. Several databases and other sources were searched to identify in vitro and animal studies, and clinical trials investigating elderberry fruit preparations. For the latter, the level of evidence was evaluated as described previously. Elderberry fruit preparations may provide antioxidant, antiviral and antiproliferative effects in vitro. One animal experiment and one clinical trial were able to back the antioxidative impact in terms of a weak antilipidemic effect. Antibacterial and antiinflammatory effects seem possible, but need further support. In rats, an aqueous elderberry fruit extract produced central depression and analgesia and an ethanol fruit extract improved acetic acid-induced colitis. Several in vitro studies together with two exploratory studies in humans and one open study in chimpanzees indicate that the aqueous elderberry extract Sambucol may be useful for the treatment of viral influenza infections. These promising effects of elderberry fruit preparations from experimental and clinical studies should be backed by more rigorous studies before these preparations are recommended in the prevention of diseases and in treatment schedules. PMID- 19548291 TI - Anticholinesterase activity of 7-methoxyflavones isolated from Kaempferia parviflora. AB - The rhizome of Kaempferia parviflora or kra-chai-dum (in Thai) is used traditionally as a folk medicine. The preliminary cholinesterase inhibitory screening of this plant extract exhibited significant acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities. Thirteen known methoxyflavones (1-13) were isolated and their structures were completely elucidated based on NMR analysis and compared with literature reports. Minor compounds 12-13 were reported for the first time from this species. The cholinesterase inhibitory test results showed that the highest potential inhibitors toward AChE and BChE were 5,7,4'-trimethoxyflavone (6) and 5,7 dimethoxyflavone (7), respectively, with the percentage inhibitory activity varying over 43-85%. The structure-activity relationship study led to the conclusion that compounds bearing 5,7-dimethoxy groups and a free substituent at C-3 had a significant inhibitory effect at a concentration of 0.1 mg/mL, but those bearing a 5-hydroxyl group reduced the inhibitory potency. On the other hand, flavones bearing a 3'- or 5'-methoxy group did not influence the inhibitory effect. Interestingly, 5,7-dimethoxyflavone (7) exhibited strong selectivity for BChE over AChE which may be of great interest to modify as a treatment agent for Alzheimer's disease. PMID- 19548292 TI - Efficacy of grain protectants against four psocid species on maize, rice and wheat. AB - BACKGROUND: Psocids are emerging pests in stored products, particularly in amylaceous commodities such as grains. Currently, their control is based on the use of fumigants and contact insecticides; however, newer data indicate that psocids are tolerant to insecticides used to control other stored-grain species. This study evaluated the insecticides registered in the USA for use on stored maize, rice and wheat for control of the psocid species Lepinotus reticulatus, Liposcelis entomophila, L. bostrychophila and L. paeta. Mortality of exposed adult females was recorded after 7 and 14 days of exposure, while progeny production was assessed after 30 days of exposure. RESULTS: On wheat and rice, chlorpyriphos-methyl + deltamethrin was generally more effective against exposed parental adults than spinosad or pyrethrin, while pirimiphos-methyl was more effective on maize than spinosad or pyrethrin. In most cases, progeny production was suppressed in the treated grains. Progeny production was consistently lowest on wheat and rice treated with chlorpyriphos-methyl + deltamethrin and maize treated with pirimiphos-methyl. CONCLUSIONS: Chlorpyriphos-methyl + deltamethrin and pirimiphos-methyl were the most effective insecticides for all species and commodities. Conversely, efficacy of spinosad or pyrethrum was highly dependent on the psocid species and commodity. PMID- 19548293 TI - Esterase-mediated resistance to pyrethroids in field populations of Helicoverpa armigera (Lepidoptera: Noctuidae) from Central Africa. AB - BACKGROUND: Evolution of pyrethroid resistance in the cotton bollworm Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) threatens continued cotton production in Central Africa. Dose-response bioassays were conducted on area-wide collection of bollworm populations from major host plants, while biochemical techniques were used to evaluate basic mechanisms underlying resistance. RESULTS: Pyrethroid resistance is primarily associated with detoxification by enhanced esterase activity. High resistance to cypermethrin (RF = 67-1771), cross-resistance to deltamethrin (RF = 60-2972) and lack of cross-resistance to the non-ester pyrethroid etofenprox (RF = 2-10) were observed among H. armigera field populations and laboratory-selected strains. Enzymatic assays showed that esterase activity, but not oxidase content or glutathione-S-transferase activity, was positively correlated with resistance to cypermethrin. Pretreatment with piperonyl butoxide (PBO) resulted in significant synergism with cypermethrin in 6/10 field populations, but not in the laboratory-selected strain, indicating that additional mechanisms such as mixed-function oxidase (MFO) may be involved in field resistance. The absence of cross-resistance to DDT ruled out a possible target-site modification. CONCLUSION: Knowledge of the mechanisms involved in pyrethroid resistance and the lack of cross-resistance to spinosad and indoxacarb is a key to devising new resistance management strategies aimed at restoring the efficacy of pyrethroid-based programmes. PMID- 19548295 TI - Resonance balance shift in stacks of delocalized singlet biradicals. PMID- 19548294 TI - Duloxetine in the treatment of irritable bowel syndrome: an open-label pilot study. AB - OBJECTIVE: To assess the efficacy of duloxetine for irritable bowel syndrome (IBS). METHODS: We conducted an open-label 12-week trial of duloxetine 60 mg daily in 15 patients with IBS without concurrent major depressive disorder. The primary outcome measure was average abdominal pain. Secondary measures included IBS symptoms, Clinical Global Impression-Severity, Hamilton Anxiety Rating Scale, IBS Quality-of-Life Scale, and Sheehan Disability Scale. We analyzed changes using random regression and one-sample t-tests. RESULTS: Fourteen patients completed at least one post-baseline evaluation; eight completed the study. Duloxetine was associated with significant improvement (p < 0.05) in pain, severity of illness, quality of life, loose stool, work and family disability, and anxiety. However, duloxetine did not improve hard stool. Although we found no evidence of serious duloxetine toxicity, seven participants withdrew over the course of the study because of adverse drug events. CONCLUSIONS: In this small, open-label study, duloxetine appeared to be effective for many features of IBS, but its adverse effects, most notably constipation, limited its use. Since our study excluded individuals with concurrent major depression, it appears that duloxetine may benefit IBS independently of its antidepressant effects. These encouraging but preliminary open-label findings support further investigation of duloxetine treatment in placebo-controlled trials of IBS. PMID- 19548297 TI - Diazinon reduction and partitioning between water, sediment and vegetation in stormwater runoff mitigation through rice fields. AB - BACKGROUND: Contamination of surface waters by pesticides is a concern in the United States and around the world. Innovative mitigation strategies are needed to remediate this potential environmental contaminant. One potential solution is to divert pesticide-laden drainage or surface water through agricultural rice fields. With a hydroperiod, hydrosoil and hydrophyte (rice), these systems serve essentially as a type of constructed wetland. In both summer and fall experiments, diazinon-amended water was diverted through two rice ponds at the University of Mississippi Field Station. Likewise, a non-vegetated control pond was amended with diazinon-laden water. Water, sediment and plant samples were taken spatially and temporally to determine the distribution of diazinon within systems. RESULTS: Outflow diazinon concentrations decreased significantly (P < 0.05) from inflow in both vegetated ponds for both preharvest and post-harvest experiments. Although sorption to rice plants was minimal in the overall mass distribution of diazinon (1-3%), temporal data indicated that diazinon concentrations reached the outflow sediment of the non-vegetated control twice as fast as in either vegetated (rice) system. In both vegetated systems, sediment diazinon concentrations decreased (77 and 100%) from inflow to outflow, while a decrease of <2% was noted in the non-vegetated control. CONCLUSIONS: Diversion of pesticide-contaminated water through rice fields demonstrated potential as a low cost, environmentally efficient mitigation practice. Studies on these systems are continuing to evaluate the optimal chemical retention time for rice field mitigation, as well as diazinon transfer to rice grain seeds that may be used as a food source. PMID- 19548298 TI - Multivariate statistical analysis applied to an IL6 signal transduction model in hepatocytes. AB - This paper introduces the application of linear multivariate statistical techniques, including partial least squares (PLS), canonical correlation analysis (CCA) and reduced rank regression (RRR), into the area of Systems Biology. This new approach aims to extract the important proteins embedded in complex signal transduction pathway models. The analysis is performed on a model of intracellular signalling along the janus-associated kinases/signal transducers and transcription factors (JAK/STAT) and mitogen activated protein kinases (MAPK) signal transduction pathways in interleukin-6 (IL6) stimulated hepatocytes, which produce signal transducer and activator of transcription factor 3 (STAT3). A region of redundancy within the MAPK pathway that does not affect the STAT3 transcription was identified using CCA. This is the core finding of this analysis and cannot be obtained by inspecting the model by eye. In addition, RRR was found to isolate terms that do not significantly contribute to changes in protein concentrations, while the application of PLS does not provide such a detailed picture by virtue of its construction. This analysis has a similar objective to conventional model reduction techniques with the advantage of maintaining the meaning of the states prior to and after the reduction process. A significant model reduction is performed, with a marginal loss in accuracy, offering a more concise model while maintaining the main influencing factors on the STAT3 transcription. The findings offer a deeper understanding of the reaction terms involved, confirm the relevance of several proteins to the production of Acute Phase Proteins and complement existing findings regarding cross-talk between the two signalling pathways. PMID- 19548299 TI - A unified approach for analyzing exchangeable binary data with applications to developmental toxicity studies. AB - In this article, we present a general procedure to analyze exchangeable binary data that may also be viewed as realizations of binomial mixtures. Our approach unifies existing models and is practical and computationally easy. Resulting from completely monotonic functions, we introduce a rich family of parametric parsimonious binomial mixtures, including the incomplete Beta-, Gamma-, Normal-, and Poisson-binomial, generalizing the Beta-binomial. We show that the family is closed under convex linear combinations, products, and composites. We also give the moments and the Markov property of the family. With such distributions, we can perform statistical inference on correlated binary data and, in particular, overdispersed data. We propose a regression procedure that generalizes logistic regression. We provide a forward model selection procedure. We run a small simulation to validate the inclusion of the binomial distribution. Finally, we apply the proposed procedure to analyze the 2, 4, 5-Trichlorophenoxyacetic acid and E2 data and compare the results with existing procedures. PMID- 19548300 TI - Purification and characterisation of proteins secreted by the entomopathogenic fungus Metarhizium anisopliae with insecticidal activity against adults of the Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae). AB - BACKGROUND: The control of the Mediterranean fruit fly (medfly) Ceratitis capitata (Wied) is usually performed with protein bait sprays incorporating chemical insecticides that may have adverse effects on humans, non-target organisms and the environment. In recent years, scientists have sought more environmentally friendly insecticides for medfly control, such as plant- and microorganism-derived compounds. Among these compounds, entomopathogenic fungi are an unexplored source of natural insecticides. RESULTS: The crude soluble protein extract (CSPE) of the entomopathogenic fungus Metarhizium anisopliae (Mestch.) (strain EAMa 01/58-Su) shows chronic insecticidal activity when administered per os. Mortality in flies exhibits a dose response. The CSPE produces an antifeedant effect in adult flies, a result probably due to a progressive deterioration of the fly midgut after ingestion of the extract. Protease and temperature treatments show that insecticidal activity against C. capitata is due to proteinaceous compounds that are highly thermostable. Four monomeric proteins from this crude extract have been purified by liquid chromatography and gel electroelution. Although all four monomers seem to be involved in the insecticidal activity of the CSPE, the 15 kDa and the 11 kDa proteins appear to be mainly responsible for the observed insecticidal effect. CONCLUSIONS: Four new fungal proteins with insecticidal activity have been purified and identified. These proteins might be combined with insect baits for C. capitata biocontrol. PMID- 19548302 TI - Light and scanning electron microscopic study on the tongue and lingual papillae of the common hippopotamus, Hippopotamus amphibius amphibius. AB - We observed the three-dimensional structures of the external surface and connective tissue cores CTCs, after exfoliation of the epithelium of the lingual papillae (filiform, fungiform, and foliate papillae) of the common hippopotamus (Hippopotamus amphibius amphibius) using scanning electron microscopy and conventional light microscopy. Following unique features were found; typical vallate papillae with a circumferential furrow were not observable. Instead, numerous large fungiform papillae were rather densely distributed on the posterior of the lingual prominence. Taste buds were observable only on the dorsal epithelium. Serous lingual gland was not seen in the lamina propria; however, mucous-rich mixed lingual glands were found and in a few of orifices were seen on the large fungiform tops. Lingual prominence was diminished their width. Rather long and slender conical papillae were distributed on the lingual prominence and were similar to nonruminant herbivore, that is donkey. Beside this narrow lingual prominence, lateral slopes were situated with numerous short spine like protrusions. After removal of the epithelium, CTCs of lateral slopes exhibited attenuated flower bud structures. Large-conical papillae were situated on the root of the tongue. These large conical papillae were not seen among ruminants and seen on the lingual root of omnivores and carnivores. It implies that lingual structure of common hippopotamus possessed mixed characteristics between Perissodactyls, Ruminantia, and nonherbivores such as Suiformes because of their unique evolutionally taxonomic position. Moreover, adaptation for soft grass diet and associating easier mastication may be also affecting these mixed morphological features of the tongue. PMID- 19548303 TI - Editorial: Canadian mass spectrometry: instrumentation development. PMID- 19548305 TI - Mechanical properties of the hindlimb bones of bullfrogs and cane toads in bending and torsion. AB - When compared with most vertebrates, frogs use a novel style of jumping locomotion powered by the hindlimbs. Hindlimb bones of frogs must withstand the potentially erratic loads associated with such saltatory locomotion. To evaluate the load bearing capacity of anuran limb bones, we used three-point bending, torsion, and hardness tests to measure the mechanical properties of the femur and tibiofibula from adults of two species that use different jumping styles: explosively jumping bullfrogs (Rana (Lithobates) catesbeiana) and cyclically hopping cane toads (Bufo (Chaunus) marinus). Yield stress and strain values for R. catesbeiana and B. marinus hindlimb bones are within the range of values previously reported for other vertebrates. However, anuran hindlimb bones generally stand out as having higher yield stresses in bending than those of closely related, nonsaltatory salamanders, highlighting the importance of considering phylogenetic context in comparisons of bone functional capacity and adaptation. Stiffness values for both frog species tested were also high, which may facilitate efficient transmission of muscular forces while jumping. Elevated stiffness may also contribute to some discrepancies between determinations of bone properties via hardness versus bending tests. In comparisons between species, B. marinus bones showed significantly higher bending yield stresses than R. catesbeiana, whereas R. catesbeiana bones showed significantly higher torsional yield stresses than B. marinus. These differences may correlate with differences in jumping style and limb anatomy between ranid and bufonid frogs, suggesting that evolutionary changes in bone mechanical properties may help to accommodate new functional demands that emerge in lineages. PMID- 19548307 TI - Effects of fasting at different stages of lighting regimen on the proliferation of jejunal epithelial cells during rat pup weaning. AB - The lifespan of intestinal epithelial cells is predetermined by the process of cell proliferation that occurs constantly in the crypt. The control of this process involves some endogenous factors, such as hormones, as well as exogenous factors, like food and natural light variations. These last two exogenous factors seem to be the major modulators of the cell proliferation process. Fasting treatment was conducted to assess the role of food and its effect on the metaphase index (MI) of the intestinal epithelium at different times and periods (light and dark) of the day. The effects of short- (5 hr) and long-term (25 hr) fasting on the MI in the jejunal epithelium of young rats were investigated at 09:00 h, 15:00 hr, 21:00 hr, and 02:00 hr using the arrested metaphases method. The present study demonstrates that 5 hr and 25 hr of fasting treatment decrease the MI at 09:00 hr. It was observed from MI analysis that there is an interaction between the fed/fasted status of the animal and the different times of the day. This result suggests that during the transition from youth to adulthood, the control of MI by the light/dark cycle seems to be more pronounced as compared with control by food intake at some periods of the day, although at other times food had a greater impact on the MI. PMID- 19548306 TI - The antiproliferation effect of berbamine on k562 resistant cells by inhibiting NF-kappaB pathway. AB - Imatinib mesylate is effective against Ph chromosome-positive leukemia; however, resistance has been reported. High expression of bcr-abl in mRNA and protein levels, and other alterations were found in patients who experienced imatinib treatment failures and thus it is important to design alternative treatment strategies. The aim of this study was to evaluate the in vitro effect of berbamine, on imatinib-resistant chronic myelogenous leukemia (CML) K562 (K562-r) cells, and explore the mechanisms. The growth of K562-r cells was examined using the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Morphological analysis and DNA agarose electrophoresis were used to detect apoptosis in K562-r cells, and the extent of the cells in the sub-G1 cell cycle phase was measured using flow cytometry. The expression levels of BCR-ABL, phospho-BCR-ABL, and nuclear factor kappaB (NF-kappaB), IkappaBalpha, phospho IkappaBalpha, IkappaB kinases alpha(IKKalpha), and Survivin were determined by Western blot. bcr-abl mRNA expression was determined by RT-PCR. MTT assays indicated that berbamine significantly inhibited the proliferation of K562-r cells. Cells with characteristics of apoptosis were confirmed by morphology examination and DNA agarose electrophoresis and percentage of apoptosis were increased after treatment with berbamine. The results also showed that berbamine was able to down-regulate BCR-ABL and phospho-BCR-ABL proteins by affecting bcr abl mRNA expression and decrease expression of nuclear NF-kappaB, phospho IkappaBalpha, IKKalpha, and Survivin. Collectively, berbamine could inhibit the proliferation of K562-r cells and induce apoptosis. The mechanisms may be related at least in part, to inhibit BCR-ABL and its downstream NF-kappaB signaling. Berbamine may provide an alternative candidate for the treatment of patients with CML resistant to imatinib therapy. PMID- 19548308 TI - Plasticity of interstitial cells of cajal: a study in the small intestine of adult Guinea pigs. AB - Although it is well known that the reduction of interstitial cells of Cajal (ICCs) is associated with several gastrointestinal motility disorders in clinic, it is unknown whether the mature ICCs still have an active plasticity in adult mammals. This study focused on the issues of the reduction of ICCs during Imatinib administration and the recovery of ICCs following drug withdrawal in the small intestine of adult guinea pigs. ICCs were revealed by immunofluorescence on whole mount preparations with anti-Kit, alpha-smooth muscle actin, (alpha-SMA), and 5-bromo-2'-deoxyuridine (BrdU) antibodies. Moreover, the occurrence of apoptosis was also assayed. Imatinib treatment led to a gradual reduction of ICCs in number around the myenteric plexus and deep muscular plexus, which was dependent on the time but no apoptosis of ICCs was detected with the TUNEL method. During Imatinib treatment, some ICC-like cells were double labeled for Kit and alpha-SMA and a few ICC-like cells were only stained with alpha-SMA. When Imatinib was discontinued, the number of ICCs recovered to normal within 32 days. During this time, some proliferating ICCs were demonstrated by double labeling with Kit and BrdU antibodies. Our results indicated that Kit signaling was essential for the maintenance of survival and proliferation of the mature ICCs in the small intestine of adult guinea pigs. Moreover, ICCs might transdifferentiate to a type of alpha-SMA(+) cells, perhaps a phenotype of smooth muscle cells, when there is a loss-of-function of Kit. PMID- 19548309 TI - TGFbetaR2 aberrant methylation is a potential prognostic marker and therapeutic target in multiple myeloma. AB - Multiple myeloma (MM) is an incurable hematological malignancy. Different studies demonstrated the occurrence of genetic and epigenetic alterations in MM. The aberrant methylation is one of the most frequent epigenetic alterations in human genome. This study evaluated the aberrant methylation status of 20 genes in 51 MM samples by quantitative methylation-specific PCR (QMSP) and compared the methylation profile with clinicopathological characteristics of the patients. The QMSP analyses showed that PTGS2 (100.0%), SFN (100.0%), CDKN2B (90.2%), CDH1 (88.2%), ESR1 (72.5%), HIC1 (70.5%), CCND2 (62.7%), DCC (45.1%) and TGFbetaR2 (39.2%) are frequently hypermethylated in MM while aberrant methylation of RARbeta (16.6%), MGMT (12.5%), AIM1 (12.5%), CDKN2A (8.3%), SOCS1 (8.3%), CCNA1 (8.3%) and THBS1 (4.1%) are rare events. There was no methylation of GSTP1, MINT31, p14ARF and RB1 in the samples tested. Hypermethylation of ESR1 was correlated positively with isotype IgA, while aberrant methylation of THBS1 correlated negatively with isotype IgG. Furthermore, hypermethylation of DCC and TGFbetaR2 were correlated with poor survival. The multivariate analysis showed ISS and TGFbetaR2 hypermethylation strongly correlated with poor outcome. This study represents the first quantitative evaluation of promoter methylation in MM and our data provide evidence that TGFbetaR2 hypermethylation, besides ISS, may be useful as prognostic indicator in this disease. PMID- 19548310 TI - Heterogeneous ribonucleoprotein K is a marker of oral leukoplakia and correlates with poor prognosis of squamous cell carcinoma. AB - Oral leukoplakia is a heterogeneous lesion with risk of cancer development; there are no biomarkers to predict its potential of malignant transformation. Tissue proteomic analysis of oral leukoplakia using iTRAQ labeling liquid chromatography mass spectrometry showed overexpression of heterogeneous ribonucleoprotein K (hnRNP K), a transformation-related RNA-binding protein, in leukoplakia in comparison with normal tissue. Herein, we investigated the clinical significance of hnRNP K in identification of oral leukoplakic lesions in early stages and as a prognostic marker in head-and-neck/oral squamous cell carcinomas (HNOSCCs). Immunohistochemical analysis of hnRNP K was performed in 100 HNOSCCs, 199 leukoplakias and 55 nonmalignant tissues and correlated with clinicopathologic parameters and disease prognosis over 6 years for HNOSCCs. hnRNP K nuclear expression increased from normal tissues to leukoplakia, and frank malignancy (p < 0.001). Cytoplasmic hnRNP K increased significantly from leukoplakia to HNOSCCs (p < 0.001) and was associated with poor prognosis of HNOSCCs (p = 0.011) by Kaplan-Meier analysis. The most important finding of our follow-up study is that cytoplasmic hnRNP K is an independent predictor of disease recurrence in HNOSCC patients. In conclusion, nuclear hnRNP K may serve as a potential marker for early diagnosis, whereas its cytoplasmic accumulation can help to identify a subgroup of HNOSCC patients with poor prognosis, suggesting its putative utility in clinical management of HNOSCC. PMID- 19548311 TI - Solubility of hydrophobic compounds in water-cosolvent mixtures: relation of solubility with water-cosolvent interactions. AB - The solubility of organic compounds in mixtures of water and an organic cosolvent can be reasonably estimated from the solubility values in the neat solvents and the composition of the solvent mixture, by means of the log-linear solubilization model. However, deviations from the model are frequently observed in different degree. Such deviations, which tend to become more pronounced with decreasing polarity of the cosolvent, are to a good extent the result the nonideal mixing of water and cosolvent, due to the interactions between the two solvent components. We present a model that incorporates the effect of water-cosolvent interactions into the log-linear model. The effect of nonideality of mixing takes the form of a pseudoquadratic expression on the cosolvent concentration, obtainable from vapor pressure data of the solute-free water-cosolvent mixture. PMID- 19548312 TI - Sox17-2A-iCre: a knock-in mouse line expressing Cre recombinase in endoderm and vascular endothelial cells. AB - Sox17 encodes an SRY-related high-mobility group (HMG) box transcription factor that is essential for endoderm formation and fetal hematopoietic stem cell maintenance. In the mouse, expression of Sox17 is first observed in the extraembryonic endoderm and is subsequently seen in the definitive endoderm as well as in blood and the endothelial cells of the developing vasculature. To conditionally inactivate genes in these domains, we have targeted the Sox17 locus to generate a bicistronic mRNA linking Sox17 to a codon improved Cre recombinase (iCre) via a viral 2A sequence. Here we report a new Cre knock-in mouse line, Sox17-2A-iCre, with activity in the developing endoderm, the vascular endothelial cells of the cardiovascular system and the hematopoietic system. Our results indicate that the Sox17-2A-iCre is active in an early endoderm progenitor and recombination of the Rosa26 reporter was observed in all previously reported expression domains of Sox17. The Sox17-2A-iCre line will be an excellent tool to conditionally inactivate genes in the definitive endoderm as well as in the vasculature and hematopoietic system. PMID- 19548313 TI - Tools for the genetic analysis of germ cells. AB - Germ cells are essential for the propagation of individual species. Studies on germ cell development in mice highlight important biological paradigms. Beginning with their first appearance around embryonic day 7 (E7), germ cells undergo specific cellular changes at different stages of their embryonic and adult development. Germ cells migrate through the hind-regions of the embryo to eventually home into the developing gonads. Further differentiation and development of germ cells differ in males and females. The processes involved in germ cell development and their eventual differentiation into sperm and oocytes have been under extensive investigation in recent years. Studies on germ cells have shed light on the cellular and molecular processes involved in their specification, migration, proliferation, death, and differentiation. These studies have also revealed much about maintenance of stem cell populations and fertility. Here we review the genetic tools that are at present available to study germ cells in the mouse. PMID- 19548315 TI - Equilibrium studies of protein aggregates and homogeneous nucleation in protein formulation. AB - Shaking or heat stress may induce protein aggregates. Aggregation behavior of an IgG1 stressed by shaking or heat following static storage at 5 and 25 degrees C was investigated to determine whether protein aggregates exist in equilibrium. Aggregates were detected using different analytical methods including visual inspection, turbidity, light obscuration, size exclusion chromatography, and dynamic light scattering. Significant differences were evident between shaken and heated samples upon storage. Visible and subvisible particles (insoluble aggregates), turbidity and z-average diameter decreased whilst soluble aggregate content increased in shaken samples over time. Insoluble aggregates were considered to be reversible and dissociate into soluble aggregates and both aggregate types existed in equilibrium. Heat-induced aggregates had a denatured protein structure and upon static storage, no significant change in insoluble aggregates content was shown, whilst changes in soluble aggregates content occurred. This suggested that heat-induced insoluble aggregates were irreversible and not in equilibrium with soluble aggregates. Additionally, the aggregation behavior of unstressed IgG1 after spiking with heavily aggregated material (shaken or heat stressed) was studied. The aggregation behavior was not significantly altered, independent of the spiking concentration over time. Thus, neither mechanically stressed native nor temperature-induced denatured aggregates were involved in nucleating or propagating aggregation. PMID- 19548314 TI - Generation of mice with a conditional allele for G6pc. AB - Glucose-6-phosphatase-alpha (G6Pase-alpha or G6PC) catalyzes the hydrolysis of glucose-6-phosphate to glucose and is a key enzyme in interprandial glucose homeostasis. Mutations in the human G6PC gene, expressed primarily in the liver, kidney, and intestine, cause glycogen storage disease Type Ia (GSD-Ia), an autosomal recessive disorder characterized by a disturbed glucose homeostasis. For better understanding of the roles of G6Pase-alpha in different tissues and in pathological conditions, we have generated mice harboring a conditional null allele for G6pc by flanking Exon 3 of the G6pc gene with loxP sites. We confirmed the null phenotype by using the EIIa-Cre transgenic approach to generate mice lacking Exon 3 of the G6pc gene. The resulting homozygous Cre-recombined null mice manifest a phenotype mimicking G6Pase-alpha-deficient mice and human GSD-Ia patients. This G6pc conditional null allele will be valuable to examine the consequence of tissue-specific G6Pase-alpha deficiency and the mechanisms of long term complications in GSD-Ia. PMID- 19548317 TI - Fluorescence of unmodified oligonucleotides: A tool to probe G-quadruplex DNA structure. AB - Fluorescence of unmodified oligonucleotides has not been exploited for guanine quadruplex (G-quadruplex) characterization. We observe that G-rich sequences fluoresce more strongly than duplex or single-stranded DNA but much more weakly than fluorophores like fluorescein. This increase in the intrinsic fluorescence is not due to an increase in absorption at the excitation wavelength but rather to a change in the quantum yield. We show that unlabeled oligonucleotides that form G-quadruplexes can be differentiated on the basis of their emission spectra from similar sequences that do not contain consecutive guanines. Intermolecular quadruplexes formed by the oligonucleotides 5'-T(4)G(n)T(4)-3' (n = 4-10) display a nonlinear, but continuous, increase in emission intensity as the G content increases. The sequence 5'-GGGT-3', which has been proposed to form a monomeric quadruplex and an interlocked quadruplex (Krishnan-Ghosh et al. J Am Chem Soc 2004, 126, 11009), was compared with the similar sequence 5'-TGGG-3', the structure of which has not been characterized. Both the maximum emission intensity and the spectral shape differ for these oligonucleotides as a function of sample preparation, indicating that different types of quadruplexes form for both sequences. Our work is the first to demonstrate that the suprastructure of G rich sequences can be probed using fluorescence signatures of unmodified oligonucleotides. PMID- 19548318 TI - Relative resistance of SGK1 knockout mice against chemical carcinogenesis. AB - The serum and glucocorticoid inducible kinase SGK1 was originally cloned from mammary tumor cells. SGK1 was found to be up-regulated in a variety of tumors, but down-regulated in several distinct tumors. Thus, evidence for a role of SGK1 in tumor growth remained conflicting. According to in vitro observations, SGK1 is up-regulated by the oncogene beta-catenin and negatively regulates the proapoptotic transcription factor FOXO3a, which in turn stimulates transcription of the Bcl2-interacting mediator BIM. This study aimed to define the role of SGK1 in colon carcinoma in vivo. SGK1 knockout mice (sgk1(-/-)) and their wild type littermates (sgk1(+/+)) were subjected to chemical cancerogenesis (intraperitoneal injection of 20 mg/kg 1,2-dimethylhydrazine followed by three cycles of 30 g/L synthetic dextran sulfate sodium for 7 days). Moreover, SGK1 was silenced in HEK293 cells. FOXO3a and BIM protein abundance was determined by Western blotting and immunohistochemistry. Following chemical cancerogenesis, sgk1(-/-)mice developed significantly less colonic tumors than sgk1(+/+)mice. According to Western blotting and immunohistochemistry, SGK1 deficiency enhanced the expression of FOXO3a and BIM both, in vitro and in vivo. SGK1 deficiency counteracts the development of colonic tumors, an effect at least in part due to up-regulation of FOXO3a and BIM. PMID- 19548319 TI - Actin-dependent dynamics of keratin filament precursors. AB - Actin filament and microtubule growth characteristics are defined by their different plus and minus ends. In contrast, intermediate filaments lack this type of polarity. Yet, intermediate filament network growth occurs by selective addition of newly formed and polymerizing keratin particles at peripheral network domains thereby allowing polarized network reorganization. To examine this process at high resolution in living cells, mammary epithelium-derived, immortalized EpH4-cells were infected with retroviral cDNA constructs coding for human keratin 18-fluorescent protein hybrids. Several stable cell lines were established presenting characteristic fluorescent keratin filament (KF) networks. These cells contain particularly large and abundant lamellipodia in which nascent keratin particle dynamics are easily detected by time-lapse fluorescence microscopy. These keratin particles originate close to the plasma membrane, translocate continuously toward the cell center, and integrate end-on into the peripheral KF network. We show that this inward-directed transport relies on intact actin filaments. After treatment with the actin filament-disrupting drug cytochalasin newly polymerizing keratin assemblies still appear in the peripheral cytoplasm but remain stationary. On the other hand, nocodazole-mediated disruption of microtubules does not affect the centripetal KF precursor transport. From these and other observations a model is deduced which postulates that focal adhesion-dependent keratin polymerization occurs in forming lamellipodia and that transport of newly formed keratin particles is mediated by actin filaments until network integration. This mechanism allows extension of the KF network toward the leading edge in migrating cells and may be of relevance for tissue development and regeneration. PMID- 19548316 TI - Cholesterol and Kir channels. AB - To date, most of the major types of Kir channels, Kir2s, Kir3s, Kir4s, and Kir6s, have been found to partition into cholesterol-rich membrane domains and/or to be regulated by changes in the level of membrane cholesterol. Surprisingly, however, in spite of the structural similarities between different Kirs, effects of cholesterol on different types of Kir channels vary from cholesterol-induced decrease in the current density (Kir2 channels) to the loss of channel activity by cholesterol depletion (Kir4 channels) and loss of channel coupling by different mediators (Kir3 and Kir6 channels). Recently, we have gained initial insights into the mechanisms responsible for cholesterol-induced suppression Kir2 channels, but mechanisms underlying cholesterol sensitivity of other Kir channels are mostly unknown. The goal of this review is to present a summary of the current knowledge of the distinct effects of cholesterol on different types of Kir channels in vitro and in vivo. PMID- 19548320 TI - The HCV ARFP/F/core+1 protein: production and functional analysis of an unconventional viral product. AB - Hepatitis C virus (HCV) is an enveloped positive-strand RNA virus of the Flaviviridae family. It has a genome of about 9,600 nucleotides encoding a large polyprotein (about 3,000 amino acids) that is processed by cellular and viral proteases into at least 10 structural and nonstructural viral proteins. A novel HCV protein has also been identified by our laboratory and others. This protein- known as ARFP (alternative reading frame protein), F (for frameshift) or core+1 (to indicate the position) protein--is synthesized by an open reading frame overlapping the core gene at nucleotide +1 (core+1 ORF). However, almost 10 years after its discovery, we still know little of the biological role of the ARFP/F/core+1 protein. Abolishing core+1 protein production has no affect on HCV replication in cell culture or uPA-SCID mice, suggesting that core+1 protein is probably not important for the HCV reproductive cycle. However, the detection of specific anti-core+1 antibodies and T-cell responses in HCV-infected patients, as reported by many independent laboratories, provides strong evidence that this protein is produced in vivo. Furthermore, analyses of the HCV sequences isolated from patients with hepatocellular carcinoma and in vitro studies have provided strong preliminary evidence to suggest that core+1 protein plays a role in advanced liver disease and liver cancer. The available in vitro data also suggest that certain core function proteins may depend on production of the core+1 protein. We describe here the discovery of the various forms of the core+1 protein and what is currently known about the mechanisms of their production and their biochemical and functional properties. We also provide a detailed summary of the results of patient-based research. PMID- 19548321 TI - The ADP-dependent sugar kinase family: kinetic and evolutionary aspects. AB - Some archaea of the Euryarchaeota present a unique version of the Embden-Meyerhof pathway where glucose and fructose-6-phosphate are phoshporylated using ADP instead of ATP as the phosphoryl donor. These are the only ADP-dependent kinases known to date. Although initially they were believed to represent a new protein family, they can be classified as members of the ribokinase superfamily, which also include several ATP-dependent kinases. As they were first identified in members of the thermococcales it was proposed that the presence of these ADP dependent kinases is an adaptation to high temperatures. Later, homologs of these enzymes were identified in the genomes of mesophilic and thermophilic methanogenic archaea and even in the genomes of higher eukaryotes, suggesting that the presence of these proteins is not related to the hyperthermophilic life. The ADP-dependent kinases are very restrictive to their ligands being unable to use triphosphorylated nucleotides such as ATP. However, it has been shown that they can bind ATP by competition kinetic experiments. The hyperthermophilic methanogenic archaeon Methanocaldococcus jannaschii has a homolog of these genes, which can phosphorylate glucose and fructose-6-phosphate. For this reason, it was proposed as an ancestral form for the family. However, recent studies have shown that the ancestral activity in the group is glucokinase, and a combination of gene duplication and lateral gene transfer could have originated the two paralogs in this member of the Euryarchaeota. Interestingly, based on structural comparisons made within the superfamily it has been suggested that the ADP dependent kinases are the newest in the group. In several members of the superfamily, the presence of divalent metal cations has been shown to be crucial for catalysis, so its role in the ADP-dependent family was investigated through molecular dynamics. The simulation shows that, in fact, the metal coordinates the catalytic ensemble and interacts with crucial residues for catalysis. PMID- 19548323 TI - Regional inequality in China's health care expenditures. AB - This paper has two parts. The first part examines the regional health expenditure inequality in China by testing two hypotheses on health expenditure convergence. Cross-section regressions and cluster analysis are used to study the health expenditure convergence and to identify convergence clusters. We find no single nationwide convergence, only convergence by cluster. In the second part of the paper, we investigate the long-run relationship between health expenditure inequality, income inequality, and provincial government budget deficits (BD) by using new panel co-integration tests with health expenditure data in China's urban and rural areas. We find that the income inequality and real provincial government BD are useful in explaining the disparity in health expenditure prevailing between urban and rural areas. In order to reduce health-spending inequality, one long-run policy suggestion from our findings is for the government to implement more rapid economic development and stronger financing schemes in poorer rural areas. PMID- 19548324 TI - Effects of Rural Mutual Health Care on outpatient service utilization in Chinese village medical institutions: evidence from panel data. AB - To solve the problem of 'Kan bing nan, kan bing gui' (medical treatment is difficult to access and expensive), a Harvard-led research team implemented a community-based health insurance scheme known as Rural Mutual Health Care (RMHC) in Chinese rural areas from 2004 to 2006. Two major policies adopted by RMHC included insurance coverage of outpatient services (demand-side policy) and drug policy (supply-side policy). This paper focuses on the effects of these two policies on outpatient service utilization in Chinese village clinics. The data used in this study are from 3-year household follow-up surveys. A generalized negative binomial regression model and a Heckman selection model were constructed using panel data from 2005 to 2007. The results indicate that the price elasticities of demand for outpatient visits and per-visit outpatient expenses were -1.5 and -0.553, respectively. After implementing the supply-side policy, outpatient visits and per-visit outpatient expenses decreased by 94.7 and 55.9%, respectively, controlling for insurance coverage. These findings can be used to make recommendations to the Chinese government on improving the health care system. PMID- 19548325 TI - Alcohol use and the labor market in Uruguay. AB - This paper is one of only a few studies to examine potential labor market consequences of heavy or abusive drinking in Latin America and the first to focus on Uruguay. We analyzed data from a Uruguayan household survey conducted in 2006 using propensity score matching methods and controlling for a number of socio demographic, family, regional, behavioral health, and labor market characteristics. As expected, we found a positive association between heavy drinking and absenteeism, particularly for female employees. Counter to the findings for developed countries, our results revealed a positive relationship between heavy drinking and labor force participation or employment. This result was mostly driven by men and weakened when considering more severe measures of abusive drinking. Possible explanations for these findings are that employment leads to greater alcohol use through an income effect, that the Uruguayan labor market rewards heavy drinking, or that labor market characteristics typical of less developed countries, such as elevated safety risks or job instability, lead to problem drinking. Future research with panel data should explore these possible mechanisms. PMID- 19548326 TI - The value of informal care--a further investigation of the feasibility of contingent valuation in informal caregivers. AB - Including informal care in economic evaluations is increasingly advocated but problematic. We investigated three well-known concerns regarding contingent valuation (CV): (1) the item non-response of CV values, (2) the sensitivity of CV values to the individual circumstances of caring, and (3) the choice of valuation method by comparing willingness-to-pay (WTP) and willingness-to-accept (WTA) values for a hypothetical marginal change in hours of informal care currently provided.The study sample consisted of 1453 caregivers and 787 care recipients. Of the caregivers, 603 caregivers (41.5%) provided both WTP and WTA values, 983 (67.7%) provided at least one. Determinants of non-response were dependent on the valuation method; primary determinants were education and satisfaction with amount of informal care provided. Caregivers' mean WTP (WTA) for reducing (increasing) informal care by 1 h was euro9.13 (10.52). Care recipients' mean WTA (WTP) for reducing (increasing) informal care by 1 h was euro8.88 (euro6.85). Values were associated with a variety of characteristics of the caregiving situation; explanatory variables differed between WTP and WTA valuations. The differences between WTP and WTA valuations were small.Based on sensitivity CV appears to be a useful method to value informal care for use in economic evalations, non-response, however, remains a matter of concern. PMID- 19548327 TI - Special issue: Incentive Motivation, Conditioning, Stress, and Neuropsychiatric Disorders: A Tribute to Jane Stewart. PMID- 19548328 TI - Ahead of the pack. PMID- 19548329 TI - CDC issues guidance for HIV patients regarding H1N1 flu virus. PMID- 19548330 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. SWITCHMRK: an avoidable raltegravir disaster. PMID- 19548331 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. When should ART be initiated in patients with OIs? PMID- 19548332 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. Continued concern about abacavir and cardiovascular risk. PMID- 19548333 TI - Report from the 16th Conference on Retroviruses and Opportunistic Infections. Investigational pharmacologic boosters. PMID- 19548334 TI - Proceedings of the 12th International Conference on Invertebrate and Fish Cell Culture, June 2008, Tucson, Arizona, USA. PMID- 19548335 TI - Proceedings of the 6th International Vanadium Symposium, 17-19 July 2008, Lisbon, Portugal. PMID- 19548336 TI - OECD: Managing Parasitic Weeds. Proceedings of a conference, 21-26 September 2008, Ostuni, Italy. PMID- 19548337 TI - Retraction. Rodriguez C, Khachemoune A. Man with whitish papule on ear. J Am Acad Dermatol 2005;53:186-90. PMID- 19548338 TI - An unnecessary visit to the emergency room. PMID- 19548339 TI - Multimodality evaluation of intracranial vasodilatory reserve in carotid artery disease. PMID- 19548341 TI - The ordered transmission disequilibrium test: detection of modifier genes. AB - We consider the problem of detection of modifier genes that lead to variations in a disease-related continuous variable (DRCV), such as the age of onset or a measure of disease severity, in a strategy of candidate genes. We propose a novel method, the ordered transmission disequilibrium test (OTDT), to test for a relation between the clinical heterogeneity expressed by a DRCV and marker genotypes of a candidate gene. The OTDT applies to trio families with one patients and his parents, all three genotyped at a bi-allelic marker M. The OTDT aims to find a critical value of the DRCV which separates the sample of families in two subsamples in which the transmission rates are significantly different. We investigate the power of the method by simulations under various genetic models and covariate distributions and compare it with a linear regression analysis. PMID- 19548340 TI - Alcohol abuse and acute behavioural disturbances in a 24-year-old patient. Diagnosis: Marchiafava-Bignami disease (MBD). PMID- 19548343 TI - Matrix attachment region elements have small and variable effects on transgene expression and stability in field-grown Populus. AB - Matrix attachment regions (MARs) are thought to buffer transgenes from the influence of surrounding chromosomal sequences, and therefore to reduce transgene silencing and variation in expression. The statistical properties of more than 400 independent transgenic events produced in Populus, with and without flanking MAR elements from the tobacco root gene RB7, were analysed. The expression of two reporter genes in two poplar clones during three phases of vegetative growth, and the association of T-DNA characteristics with expression, was examined. It was found that MARs did not show a consistent effect on transgene expression levels; they had no effect on the green fluorescent protein (GFP) reporter gene, but reduced expression in the Basta resistance (BAR) reporter gene by 23%. The presence of MARs reduced expression variability within transformant populations, apparently by reducing the number of silenced or weakly expressing events. Transgene expression was highly stable over vegetative growth cycles that spanned 3 years of growth in the glasshouse and field, but MARs showed no association with the strength of correlations in expression over the years. Nonetheless, MARs increased the correlation in expression between a p35S::GFP and prbcS::BAR transgene linked on the same vector, but the effect was small and varied between the years. The presence of MARs had no effect on the transgene copy number, but was positively associated with T-DNA truncations, as well as with the formation of direct over inverted repeats at the same chromosomal locus. PMID- 19548342 TI - A joint transcriptomic, proteomic and metabolic analysis of maize endosperm development and starch filling. AB - The maize endosperm transcriptome was investigated through cDNA libraries developed at three characteristic stages: (i) lag phase [10 days after pollination (DAP)]; (ii) beginning of storage (14 DAP); and (iii) maximum starch accumulation rate (21 DAP). Expressed sequence tags for 711, 757 and 384 relevant clones, respectively, were obtained and checked manually. The proportion of sequences with no clear function decreased from 35% to 20%, and a large increase in storage protein sequences (i.e. 5% to 38%) was observed from stages (i) to (iii). The remaining major categories included metabolism (11%-13%), transcription-RNA processing-protein synthesis (13%-20%), protein destination (5% 9%), cellular communication (3%-9%) and cell rescue-defence (4%). Good agreement was generally found between category rank in the 10-DAP transcriptome and the recently reported 14-DAP proteome, except that kinases and proteins for RNA processing were not detected in the latter. In the metabolism category, the respiratory pathway transcripts represented the largest proportion (25%-37%), and showed a shift in favour of glycolysis at 21 DAP. At this stage, amino acid metabolism increased to 17%, whereas starch metabolism surprisingly decreased to 7%. A second experiment focused on carbohydrate metabolism by comparing gene expression at three levels (transcripts, proteins and enzyme activities) in relation to substrate or product from 10 to 40 DAP. Here, two distinct patterns were observed: invertases and hexoses were predominant at the beginning, whereas enzyme patterns in the starch pathway, at the three levels, anticipated and paralleled starch accumulation, suggesting that, in most cases, transcriptional control is responsible for the regulation of starch biosynthesis. PMID- 19548344 TI - High-level expression of human immunodeficiency virus antigens from the tobacco and tomato plastid genomes. AB - Transgene expression from the plant's plastid genome represents a promising strategy in molecular farming because of the plastid's potential to accumulate foreign proteins to high levels and the increased biosafety provided by the maternal mode of organelle inheritance. In this article, we explore the potential of transplastomic plants to produce human immunodeficiency virus (HIV) antigens as potential components of an acquired immunodeficiency syndrome (AIDS) vaccine. It is shown that the HIV antigens p24 (the major target of T-cell-mediated immune responses in HIV-positive individuals) and Nef can be expressed to high levels in plastids of tobacco, a non-food crop, and tomato, a food crop with an edible fruit. Optimized p24-Nef fusion gene cassettes trigger antigen protein accumulation to up to approximately 40% of the plant's total protein, demonstrating the great potential of transgenic plastids to produce AIDS vaccine components at low cost and high yield. PMID- 19548345 TI - Plastid transformation of high-biomass tobacco variety Maryland Mammoth for production of human immunodeficiency virus type 1 (HIV-1) p24 antigen. AB - Chloroplast transformation of the high-biomass tobacco variety Maryland Mammoth has been assessed as a production platform for the human immunodeficiency virus type 1 (HIV-1) p24 antigen. Maryland Mammoth offers the prospect of higher yields of intact functional protein per unit floor area of contained glasshouse per unit time prior to flowering. Two different transformation constructs, pZSJH1p24 (for the insertion of a native p24 cDNA between the rbcL and accD genes) and pZF5 (for the insertion of a chloroplast-codon-optimized p24 gene between trnfM and trnG) were examined for the production of p24. Plants generated with construct pZSJH1p24 exhibited a normal green phenotype, but p24 protein accumulated only in the youngest leaves (up to approximately 350 microg/g fresh weight or approximately 2.5% total soluble protein) and was undetectable in mature leaves. In contrast, some of the plants generated with pZF5 exhibited a yellow phenotype (pZF5-yellow) with detectable p24 accumulation (up to approximately 450 microg/g fresh weight or approximately 4.5% total soluble protein) in all leaves, regardless of age. Total protein in pZF5-yellow leaves was reduced by approximately 40%. The pZF5-yellow phenotype was associated with recombination between native and introduced direct repeat sequences of the rbcL 3' untransformed region in the plastid genome. Chloroplast-expressed p24 was recognized by a conformation-dependent monoclonal antibody to p24, and p24 protein could be purified from pZF5-yellow leaves using a simple procedure, involving ammonium sulphate precipitation and ion-exchange chromatography, without the use of an affinity tag. The purified p24 was shown to be full length with no modifications, such as glycosylation or phosphorylation, using N-terminal sequencing and mass spectrometry. PMID- 19548346 TI - Promises and pitfalls of quantitative structure-activity relationship approaches for predicting metabolism and toxicity. AB - The description of quantitative structure-activity relationship (QSAR) models has been a topic for scientific research for more than 40 years and a topic within the regulatory framework for more than 20 years. At present, efforts on QSAR development are increasing because of their promise for supporting reduction, refinement, and/or replacement of animal toxicity experiments. However, their acceptance in risk assessment seems to require a more standardized and scientific underpinning of QSAR technology to avoid possible pitfalls. For this reason, guidelines for QSAR model development recently proposed by the Organization for Economic Cooperation and Development (OECD) [Organization for Economic Cooperation and Development (OECD) (2007) Guidance document on the validation of (quantitative) structure-activity relationships [(Q)SAR] models. OECD Environment Health and Safety Publications: Series on Testing and Assessment No. 69, Paris] are expected to help increase the acceptability of QSAR models for regulatory purposes. The guidelines recommend that QSAR models should be associated with (i) a defined end point, (ii) an unambiguous algorithm, (iii) a defined domain of applicability, (iv) appropriate measures of goodness-of-fit, robustness, and predictivity, and (v) a mechanistic interpretation, if possible [Organization for Economic Cooperation and Development (OECD) (2007) Guidance document on the validation of (quantitative) structure-activity relationships [(Q)SAR] models. The present perspective provides an overview of these guidelines for QSAR model development and their rationale, as well as the promises and pitfalls of using QSAR approaches and these guidelines for predicting metabolism and toxicity of new and existing chemicals. PMID- 19548347 TI - A novel 4-oxo-2(E)-nonenal-derived modification to angiotensin II: oxidative decarboxylation of N-terminal aspartic acid. AB - 4-Oxo-2(E)-nonenal (4-ONE) is a major bifunctional electrophile derived from lipid hydroperoxides. A substantial amount of past research on 4-ONE-derived modifications to cellular macromolecules has led to the conclusion that 4-ONE is more reactive toward DNA and protein than 4-hydroxy-2(E)-nonenal (4-HNE). In this study, a novel discovery was made that 4-ONE mediates not only adduct formation but also oxidative decarboxylation of N-terminal aspartic acid on angiotensin (Ang) II and des-Ile5, His6, Pro7, and Phe8-Ang II [Ang II (1-4)]. This reaction was not mediated by other lipid peroxidation-derived aldehydes such as 4-HNE and trans-4,5-epoxy-2(E)-decenal (4,5-EDE). The initial reaction of 4-ONE with an N terminal alpha-amino group of Ang II or Ang II (1-4) resulted in the formation of a Schiff base intermediate. The resulting intermediate underwent tautomerization and decarboxylation followed by hydrolysis to provide an alpha-keto amide (pyruvamide) moiety at the N terminus of Ang II and Ang II (1-4). The structures of 4-ONE-derived pyruvamide-Ang II and -Ang II (1-4) were confirmed by matrix assisted laser desorption ionization-time-of-flight/mass spectrometry (MALDI TOF/MS) and postsource decay (PSD)-TOF/MS analyses and by comparisons to their authentic standards. The presence of a ketone group on the N terminus was confirmed by reduction with sodium borohydride, which resulted in the addition of two hydrogen atoms. Reactivity of 4-ONE toward N-terminal aspartic acid on Ang II was then compared with that of pyridoxal 5'-phosphate (PLP), a well-known aldehyde that efficiently converts N-terminal aspartic acid residue to pyruvate residue. The results indicated a rapid formation of 4-ONE-derived pyruvamide-Ang II and a higher reactivity of 4-ONE at its physiological concentration. This suggests that peptides or proteins containing N-terminal aspartic acid can readily react with lipid hydroperoxide-derived 4-ONE to form pyruvamides, which could modulate their biological functions. PMID- 19548348 TI - Genome-wide transcriptional responses to acrolein. AB - The lipid peroxidation product and environmental pollutant acrolein participates in many diseases. Because of its formation during tobacco combustion, its role in various smoking-related respiratory conditions including lung cancer has received increasing attention. As a reactive electrophile, acrolein seems likely to disrupt many biochemical pathways, but these are poorly characterized on a genome wide basis. This study used microarrays to study short-term transcriptional responses of A549 human lung cells to acrolein, with cells exposed to 100 microM acrolein for 1, 2, or 4 h prior to RNA extraction and transcription profiling. Major pathways dysregulated by acrolein included those involved in apoptosis, cell cycle control, transcription, cell signaling, and protein biosynthesis. Although HMOX1 is a widely used marker of transcriptional responses to acrolein, this gene was the sole upregulated member of the Nrf2-driven family of antioxidant response genes. Transcript levels of several members of the metallothionein class of cytoprotective metal-chelating proteins decreased strongly in response to acrolein. Other novel findings included strong and persistent upregulation of several members of the early growth response (EGR) class of zinc finger transcription factors. Real-time PCR and Western blotting confirmed strong upregulation of a key member of this family (EGR-2), the DNA damage response gene GADD45beta, the heat shock response participant Hsp70, and also HMOX1. Consistent with changes in Nur77 mRNA levels during the microarray study, Western blotting confirmed strong Nur77 induction at the protein level, raising the possibility that this death-inducing protein contributes to the loss of cell viability during acrolein exposure. Collectively, the transcriptional response to acrolein is complex and dynamic, with future work needed to determine whether acrolein-responsive genes identified in this study contribute to cell and tissue injury in the smoke-exposed lung. PMID- 19548349 TI - Preparation and properties of lithium and sodium peroxynitrite. AB - We describe the preparation of aqueous solutions of LiONOO and NaONOO from (Me4N)ONOO. An aqueous solution of analytically pure, commercially available (Me4N)ONOO is applied to an Amberlyst 15 column at 4 degrees C, and the Me4N+ is rapidly (in 20 min) exchanged against Li+ or Na+. The exchanged peroxynitrite is produced in yields of ca. 90% with a nitrite content of 5-6%. PMID- 19548350 TI - Effect of intestinal glucuronidation in limiting hepatic exposure and bioactivation of raloxifene in humans and rats. AB - Raloxifene (Evista) is a second generation selective estrogen receptor modulator used in the treatment of osteoporosis and for chemoprevention of breast cancer. It is bioactivated to reactive intermediates, which covalently bind to proteins and form GSH conjugates upon incubation with NADPH and GSH-supplemented human and rat liver microsomes. Despite these in vitro findings, no major raloxifene related toxic events have been reported upon its oral administration to humans. This disconnect between safety of raloxifene and its in vitro bioactivation is attributed to its presystemic metabolism via glucuronidation. Current studies investigated the effect of hepatic and intestinal glucuronidation in modulating hepatic availability of raloxifene and its subsequent bioactivation, in vitro. The study design involved preincubation of raloxifene with intestinal microsomes followed by a sequential incubation with liver microsomes. The degree of bioactivation of raloxifene was assessed from the percentage of GSH conjugate formed in liver microsomal incubations or the amount of covalent binding of raloxifene-related material to liver microsomal proteins. The results indicated that human intestinal glucuronidation limited the hepatic exposure of raloxifene that underwent bioactivation in the liver. Similar experiments with rat microsomal preparations showed very little effect of intestinal glucuronidation. This effect of intestinal glucuronidation and the observed species difference were explained by comparing the efficiency (Cl(int)) of glucuronidation and oxidation in the two species. These findings suggested that even though the rate of bioactivation in the two species was similar, the Cl(int) of glucuronidation was 7.5-fold higher in the human intestine as compared to rats. These results support the hypothesis that intestinal glucuronidation modulates the amount of raloxifene undergoing bioactivation by liver and corroborate the importance of assessing other competitive metabolic pathways and species differences in metabolism prior to extrapolation of bioactivation results from rats to humans. PMID- 19548352 TI - In vitro and in silico characterization of peroxiredoxin 6 modified by 4 hydroxynonenal and 4-oxononenal. AB - Peroxiredoxin 6 (PRX6) belongs to the 1-Cys class of peroxiredoxins and is recognized as an important antioxidant protein in tissues such as cardiac muscle, skin, and lung. Preliminary in vivo proteomic data have revealed that PRX6 is adducted by 4-hydroxynonenal (4HNE) in the livers of rats chronically fed an ethanol-containing diet. The goals of this study were to evaluate the in vitro effect of aldehyde adduction on PRX6 peroxidase activity, identify specific sites of aldehyde modification using mass spectrometry, and predict conformational changes due to adduction using molecular modeling. PRX6 was found to be resistant to inactivation via aldehyde modification; however, Western blots of adducted protein revealed that both 4HNE and 4-oxononenal (4ONE) caused extensive cross linking, resulting in high molecular mass species. Tandem mass spectrometry (ESI LC-MS/MS) analysis demonstrated multiple sites of modification, but adduction of the active site Cys47 was not observed. Molecular modeling simulations indicated that adduction at Cys91 results in a change in protein active site conformation, which potentially restricts access of 4-HNE to Cys47. The Cys91-Lys209 cross linked adducts could provide the conformational changes required to inactivate the protein by either restricting access to electrophiles or preventing important amino acid interactions within the catalytic triad. PMID- 19548351 TI - Serotonergic neurotoxic thioether metabolites of 3,4 methylenedioxymethamphetamine (MDMA, "ecstasy"): synthesis, isolation, and characterization of diastereoisomers. AB - 3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is a synthetic recreational drug of abuse that produces long-term toxicity associated with the degeneration of serotonergic nerve terminals. In various animal models, direct administration of MDMA into the brain fails to reproduce the serotonergic neurotoxicity, implying a requirement for the systemic metabolism and bioactivation of MDMA. Catechol-thioether metabolites of MDMA, formed via oxidation of 3,4 dihydroxymethamphetamine and 3,4-dihydroxyamphetamine (HHMA and HHA) and subsequent conjugation with glutathione (GSH), are selective serotonergic neurotoxicants when administered directly into brain. Moreover, following systemic administration of MDMA, the thioether adducts are present in rat brain dialysate. MDMA contains a stereogenic center and is consumed as a racemate. Interestingly, different pharmacological properties have been attributed to the two enantiomers, (S)-MDMA being the most active in the central nervous system and responsible for the entactogenic effects, and most likely also for the neurodegeneration. The present study focused on the synthesis and stereochemical analysis of the neurotoxic MDMA thioether metabolites, 5-(glutathion-S-yl)-HHMA, 5-(N-acetylcystein-S-yl)-HHMA, 2,5-bis-(glutathion-S-yl)-HHMA, and 2,5-bis-(N acetylcystein-S-yl)-HHMA. Both enzymatic and electrochemical syntheses were explored, and methodologies for analytical and semipreparative diastereoisomeric separation of MDMA thioether conjugates by HPLC-CEAS and HPLC-UV, respectively, were developed. Synthesis, diastereoisomeric separation, and unequivocal identification of the thioether conjugates of MDMA provide the chemical tools necessary for appropriate toxicological and metabolic studies on MDMA metabolites contributing to its neurotoxicity. PMID- 19548353 TI - Interaction of polycyclic aromatic hydrocarbons with human cytochrome P450 1B1 in inhibiting catalytic activity. AB - Eleven polycyclic aromatic hydrocarbons (PAHs) and 14 acetylenic PAHs and biphenyls were used to analyze interactions with cytochrome P450 (P450) 1B1 in inhibiting catalytic activity, using 7-ethoxyresorufin O-deethylation (EROD) as a model reaction. Most of the chemicals examined were direct inhibitors of P450 1B1 except for 4-(1-propynyl)biphenyl, a mechanism-based inhibitor. In the case of direct inhibition of EROD activity {15 of 24 chemicals, e.g., benzo[a]pyrene, 1 (1-propynyl)pyrene, and 3-(1-propynyl)phenanthrene}, restoration of the EROD activity occurred with increasing incubation time, and kinetic analysis showed that EROD K(m) values were higher with these inhibitors at initial stages of incubation but became lower with increasing incubation time. With the other nine chemicals, the K(m) values for P450 1B1-mediated EROD increased during the incubations. Acetylenic inhibitors, but not the 11 PAHs, induced reverse type I spectral changes with P450 1B1, and the low dissociation constants (K(s)) suggested a role for such interaction in the inhibition of catalytic activity. Studies of quenching of P450 1B1-derived fluorescence with inhibitors demonstrated that acetylenic inhibitors and PAHs interacted rapidly with P450 1B1, with K(d) values < 10 microM. However, studies of quenching of inhibitor derived fluorescence with P450 1B1 showed these interactions to be different, that is, B[a]P interacted with P450 1B1 more slowly. Molecular docking of P450 1B1, based on P450 1A2 crystal structure, suggested that there are clear differences in the interaction of PAH inhibitors with P450 1B1 and 1A2 and that these differences may explain why PAH inhibitors inhibit P450 1 enzymes by different mechanisms. The results suggest that P450 1B1 interacts with synthetic polycyclic aromatic acetylenes and PAHs in different ways, depending on the chemicals, and that these differences in interactions may explain how these chemicals inhibit P450 activities by different mechanisms. PMID- 19548354 TI - Pathways for the decay of organic dichloramines and liberation of antimicrobial chloramine gases. AB - When neutrophils phagocytose bacteria, they generate the cytotoxic agent hypochlorous acid (HOCl). The specific role that HOCl plays in bacterial killing is unclear. In the phagosome, it should react with neutrophil proteins to form protein chloramines and dichloramines. We investigated the stability of model dichloramines that are likely to be formed on N-terminal amino acids and Lys residues of proteins contained within phagosomes. Dichloramines were much more unstable than their analogous monochloramines. The stability was affected by substituents on the alpha-carbon. Amino acid dichloramines were extremely unstable, indicating that an alpha-carboxyl group facilitated decomposition. In general, the absence of a substituent enhanced stability. The carboxyl group on N terminal Glu residues favored break down, but this effect was not apparent with Asp residues. Unstable dichloramines that contained a substituent on their alpha carbon were cytotoxic and killed 50% of 10(5) Staphylococcus aureus (LD50) at a dose of approximately 2.5 nmol. Their cytotoxicity declined with time. The dichloramines of N-alpha-acetyl Lys and taurine were not bactericidal up to 10 nmol per 10(5) S. aureus. None of the analogous monochloramines were cytotoxic at this dose. Dichloramines decomposed to yield chlorimines, aldehydes, and the inorganic gases ammonia monochloramine (NH2Cl) and ammonia dichloramine (NHCl2). The LD50 values were determined for NH2Cl (0.37 +/- 0.14 nmol), NHCl2 (0.08 +/- 0.02 nmol), and HOCl (0.14 +/- 0.04 nmol). Stable products formed during the breakdown of dichloramines were not bactericidal. We propose a potential antimicrobial mechanism that explains in part how HOCl can react mainly with neutrophil components but still promote killing of phagocytosed bacteria. HOCl produced in phagosomes will react with amine groups on neutrophil proteins to form unstable dichloramines that will liberate cytotoxic NH2Cl and NHCl2. These gases will contribute to killing of ingested bacteria. PMID- 19548355 TI - Effect of feeding nitrite, ascorbate, hemin, and omeprazole on excretion of fecal total apparent N-nitroso compounds in mice. AB - It was proposed that colon cancer induced by red and nitrite-preserved meat is due to meat-derived N-nitroso compounds in the colonic contents. To explore this view, we previously showed that feeding beef and hot dogs increased the fecal output of total apparent N-nitroso compounds (ANC) in mice. In the current project, adult Swiss mice were fed a semipurified diet and water containing additives for 7 days. Feces from individual mice was collected on day 7, dried, and extracted with water. Extracts were analyzed for ANC as before. Feeding 2.0 g sodium nitrite (NaNO2)/L drinking water raised fecal ANC levels from 5 to 63 nmol/g feces. In a dose-response study, fecal ANC levels were proportional to the nitrite concentration squared. Even 32 mg NaNO2/L raised fecal ANC levels 2.3 fold (P < 0.05). In other results, 64, 125, and 250 mg hemin/kg diet, fed with 2 g NaNO2/L water, showed 2.3, 2.2, and 4.6 times the ANC level for nitrite alone. Sodium nitrate (12 g/L water) did not affect fecal ANC output. Omeprazole (400 mg/kg diet) and sodium ascorbate (23 g/kg diet), when fed with 1 g NaNO2/L water, lowered fecal ANC levels by 65 and 41%, indicating that, when nitrite was fed, acid-catalyzed reactions in the stomach produced ANC, which passed down the gut to the feces. Tests indicated that nitrosothiols constituted about 20% of fecal and hot dog ANC. The observed effect of NaNO2 is thus far not consistent with the proposed hypothesis. The enhancement by hemin may help explain why red meat is a cause of colon cancer. PMID- 19548356 TI - Role of catechin quinones in the induction of EpRE-mediated gene expression. AB - In the present study, the ability of green tea catechins to induce electrophile responsive element (EpRE)-mediated gene expression and the role of their quinones in the mechanism of this induction were investigated. To this end, Hepa1c1c7 mouse hepatoma cells were used, stably transfected with a luciferase reporter gene under the expression regulation of an EpRE from the human NAD(P)H:quinone oxidoreductase 1 (NQO1) gene. The results obtained show that several, but not all, catechins tested are able to induce EpRE-mediated gene transcription, with epigallocatechin gallate (EGCG) and gallocatechin gallate (GCG), both containing a pyrogallol and a galloyl moiety, being the most powerful inducers. Moreover, it was demonstrated that the EpRE-mediated response to catechins was increased in cells with reduced cellular glutathione (GSH) levels and decreased in cells with increased levels of GSH, corroborating a role for catechin quinones. The intrinsic capacity of catechins to form quinone type metabolites upon their oxidation was demonstrated using incubations of epigallocatechin (EGC) and EGCG with tyrosinase and the GSH-trapping method. Glutathione conjugates formed in these incubations were identified as 2'-glutathionyl-EGC, 2',6'-diglutathionyl EGC, 2'-glutathionyl-EGCG, and 2',6'-diglutathionyl-EGCG, supporting the formation of quinone type metabolites involving especially the pyrogallol moiety of these catechins. Formation of the EGCG-quinone-glutathionyl adducts was also observed in the EpRE-LUX cellular system. This further supports the importance of the pyrogallol moiety for the quinone chemistry of the catechins. Finally, the presence of the pyrogallol moiety in the catechins also results in a relatively lower half-wave oxidation potential (E1/2) and calculated heat of formation (DHF) for conversion of the catechins to their corresponding quinones, pointing at an increased ability to become oxidized. Altogether, our studies reveal that catechins, especially those containing a pyrogallol moiety, induce EpRE-mediated detoxifying gene expression and that this induction is likely to be the result of their quinone chemistry. PMID- 19548357 TI - Reversibility of covalent electrophile-protein adducts and chemical toxicity. AB - The biotin-tagged electrophiles 1-biotinamido-4-(4'-[maleimidoethylcyclohexane] carboxamido)butane (BMCC) and N-iodoacetyl-N-biotinylhexylenediamine (IAB) have been used as model electrophile probes in complex proteomes to identify protein targets associated with chemical toxicity. Whereas IAB activates stress signaling and apoptosis in HEK293 cells, BMCC does not. Cysteine Michael adducts formed from BMCC and nonbiotinylated analogues rapidly disappeared in the intact cells, whereas the adducts were stable in BMCC-treated subcellular fractions, even in the presence of the cellular reductants reduced glutathione, NADH, and NADPH. In contrast, cysteine thioether adducts formed from IAB and its nonbiotinylated analogues were stable in intact cells. Loss of the BMCC adduct in cells was reduced at 4 degrees C, which suggests the involvement of a metabolic process in adduct removal. Model studies with a glutathione-BMCC conjugate indicated rapid hydrolysis of the adducted imide ring, but neither the conjugate nor its hydrolysis product dissociated to release the electrophile in neutral aqueous buffer at significant rates. The results suggest that low BMCC toxicity reflects facile repair that results in transient adduction, which fails to trigger damage signaling pathways. PMID- 19548359 TI - Plant catechols and their S-glutathionyl conjugates as antinitrosating agents: expedient synthesis and remarkable potency of 5-S-glutathionylpiceatannol. AB - With a view to elucidating the structural requisites for effective antinitrosating properties in plant polyphenolics and their metabolites, we have undertaken a comparative investigation of the nitrite scavenging effects of representative catechol derivatives of dietary relevance in the 2,3 diaminonaphthalene (DAN) nitrosation and tyrosine nitration assays. Compounds tested included caffeic acid (1), chlorogenic acid (2), piceatannol (3), hydroxytyrosol (4), and the corresponding S-glutathionyl conjugates 5-8, which were prepared using either tyrosinase (5 and 6) or a novel, o-iodoxybenzoic acid (IBX)-based oxygenation/ conjugation methodology (7b and 8). In the DAN nitrosation assay at pH 4.0, the rank order of inhibitory activities was found to be 5-S-glutathionylpiceatannol (7b) > 3 > 1 > 2 > 2-S-glutathionylcaffeic acid (5) > 2-S-glutathionylchlorogenic acid (6) > 4 approximately 5-S glutathionylhydroxytyrosol (8). Quite unexpectedly, in the tyrosine nitration assay in 0.5 M HCl, 2 was the most efficient inhibitor followed by 1 > 4 > 3 > 7b approximately 5 > 8 > 6. Under the assay conditions, the glutathionyl conjugates were usually consumed at faster rates than the parent catechols (decomposition rates: 3 > 1 > 4 > 2). The 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) assay indicated that the most effective hydrogen donors were 4 > 7b > 1 approximately 3. Overall, these results indicated that catechol compounds and their glutathionyl conjugates may exhibit profoundly different inhibitory properties depending on the specific conditions of the assay, including especially pH, and that their antinitrosating properties do not correlate tout-court with their hydrogen donor capacity. The glutathionyl-piceatannol conjugate 7b was found to be one of the most potent inhibitors in the physiologically relevant DAN assay and may provide a new structural lead for the design of effective antinitrosating agents based on dietary polyphenolic compounds. PMID- 19548358 TI - Comparison of in vitro bioactivation of flutamide and its cyano analogue: evidence for reductive activation by human NADPH:cytochrome P450 reductase. AB - Flutamide (FLU), a nonsteroidal antiandrogen drug widely used in the treatment of prostate cancer, has been associated with idiosyncratic hepatotoxicity in patients. It is proposed that bioactivation of FLU and subsequent binding of reactive metabolite(s) to cellular proteins play a causative role. A toxicogenomic study comparing FLU and its nitro to cyano analogue (CYA) showed that the nitroaromatic group of FLU enhanced cytotoxicity to hepatocytes, indicating that reduction of the nitroaromatic group may represent a potential route of FLU-induced hepatotoxicity [Coe et al. (2007) Chem. Res. Toxicol. 20, 1277-1290]. In the current study, we compared in vitro bioactivation of FLU and CYA in human liver microsomes and cryopreserved human hepatocytes. A nitroreduction metabolite FLU-6 was formed in liver microsomal incubations of FLU under atmospheric oxygen levels and, to a greater extent, under anaerobic conditions. Seven glutathione (GSH) adducts of FLU, FLU-G1-7, were tentatively identified in human liver microsomal incubations using liquid chromatography tandem mass spectrometry (LC/ MS/MS), while CYA formed only four corresponding GSH adducts, CYA-G1-4, under the same conditions. Of particular interest was the formation of FLU-G5-7 from FLU, where the nitroaromatic group of FLU was reduced to an amino group. A tentative pathway is that upon nitroreduction, the para diamines undergo cytochrome P450 (P450)-catalyzed two-electron oxidations to form corresponding para-diimine intermediates that react with GSH to form GSH adducts FLU-G5-7, respectively. The identities of FLU-G5-7 were further confirmed by LC/MS/MS analyses of microsomal incubations of a synthesized standard FLU-6. In an attempt to identify enzymes involved in the nitroreduction of FLU, NADPH:cytochrome P450 reductase (CPR) was shown to reduce FLU to FLU-6 under both aerobic and anaerobic conditions. Furthermore, the formation of FLU-G5-7 was completely blocked by the addition of a reversible CPR inhibitor, alpha-lipoic acid, to the incubations of FLU under aerobic conditions. In summary, these results clearly demonstrate that nitroreduction of FLU by CPR contributes to bioactivation and potentially to hepatotoxicity of FLU. PMID- 19548360 TI - Notopterygium forbesii boiss extract and its active constituents increase reactive species and heme oxygenase-1 in human fetal hepatocytes: mechanisms of action. AB - Notopterygium forbesii Boiss (NF) has been used as a traditional Chinese medicine for the treatment of common cold and rheumatism. However, there has been limited research on the biological properties of NF, and the mechanisms of action remain unknown. Here, we aimed to study the mechanism of NF-induced heme oxygenase-1 (HO 1) in human fetal hepatocytes (HFHs) and to identify the constituents responsible. Exposure of HFHs to NF causes oxidative stress with the accumulation of reactive species, which in turn leads to the phosphorylation of p38 MAPK and nuclear accumulation of Nrf2 transcription factor, and eventually increased levels of HO-1 mRNA and protein. The increases in reactive species and HO-1 protein are inhibited by agonists of glucocorticoid receptors (GR), such as RU28362, prednisolone, and dexamethasone, as well as by N-acetyl-L-cysteine and SB203580 (a p38 inhibitor), suggesting a role of GR in NF-induced increases in reactive species and HO-1. Assay-guided fractionation of NF led to three active compounds, phenethyl ferulate, bergaptol, and isoimperatorin, that were found to increase oxidative stress and HO-1 protein levels in HFHs. The induction of HO-1 protein in response to moderate oxidative stress may explain some of the beneficial pharmacological effects of NF. PMID- 19548362 TI - HCV, HBV and HIV in drug addicts. PMID- 19548363 TI - Fleeting or pandemic? RNs draw on experience, planning to respond to current influenza outbreak. PMID- 19548361 TI - Effects of acrolein on leukotriene biosynthesis in human neutrophils. AB - Acrolein is a toxic, highly reactive alpha,beta-unsaturated aldehyde that is present in high concentrations in cigarette smoke. In the current study, the effect of acrolein on eicosanoid synthesis in stimulated human neutrophils was examined. Eicosanoid synthesis in neutrophils was initiated by priming with granulocyte-macrophage colony-stimulating factor (GM-CSF) and subsequent stimulation with formyl-methionyl-leucyl-phenylalanine (fMLP) and 5-lipoxygenase (5-LO) products in addition to small amounts of cyclooxygenase (COX) products were detected using LC/MS/MS. A dose-dependent decrease in the formation of 5-LO products was observed in GM-CSF/fMLP-stimulated neutrophils when acrolein (0-50 microM) was present with almost complete inhibition at > or = 25 microM acrolein. The production of COX products was not affected by acrolein in these cells. The effect of acrolein was examined on key parts of the eicosanoid pathway, such as arachidonic acid release, intracellular calcium ion concentration, and adenosine production. In addition, the direct effect of acrolein on 5-LO enzymatic activity was probed using a recombinant enzyme. Some of these factors were affected by acrolein but did not completely explain the almost complete inhibition of 5-LO product formation in GM-CSF/fMLP-treated cells with acrolein. In addition, the effect of acrolein on different stimuli that initiate the 5-LO pathway [platelet activating factor (PAF)/fMLP, GM-CSF/PAF, opsonized zymosan, and A23187] was examined. Acrolein had no significant effect on the leukotriene production in neutrophils stimulated with PAF/fMLP, GM-CSF/ PAF, or OPZ. Additionally, 50% inhibition of the 5-LO pathway was observed in A23187-stimulated neutrophils. Our results suggest that acrolein has a profound effect on the 5-LO pathway in neutrophils, which may have implications in disease states, such as chronic obstructive pulmonary disease and other pulmonary disease, where both activated neutrophils and acrolein are present. PMID- 19548364 TI - Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants. Preface. PMID- 19548365 TI - When operating is considered futile: difficult decisions in the neonatal intensive care unit. PMID- 19548366 TI - Abstracts of the 38th National Conference of Immunology. September 24-26, 2008. PMID- 19548367 TI - A nested case-control study of female breast cancer in Karunagappally cohort in Kerala, India. AB - Lifestyle factors related to breast cancer risk were examined in a case-control study nested in a cohort in Karunagappally, Kerala, South India. We sought interviews with all the residents in Karunagappally with the population of 385,103 (191,149 males and 193,954 females) in the 1991 census and established a cohort of 359,619 (93% of the population in 1991) in 1990. For analysis 264 breast cancer cases with age > or = 20 years were selected from 438 breast cancer cases reported during the period 1990-2004 and for each case 3 non-cancer controls were randomly selected matched for age, religion and place of residence through the Cancer Registry, Karunagappally. Conditional logistic regression was used for the analysis. In the present study, in addition to a low number of pregnancies (P <0.001 and P for trend <0.001), more frequent intake of roots and tubers except tapioca (cassava) (OR for > or = 5 times =1.56, 95% CI=1.09, 3.09, P for trend <0.05), milk drinking (OR=1.78, 95% CI=1.17-2.69, P<0.01) and consumption of chicken meat (OR=1.84, 95% CI=1.09-3.09, P<0.05) were found to increase breast cancer risk. The present study further showed that consumption of tapioca which is a commonly used food item in South India, particularly in Kerala, reduced breast cancer risk (OR=0.55, 95% CI=0.37-0.83, P<0.01). Risk analysis was attempted among pre- and post-menopausal women separately and similar odds ratio were obtained. Consumption of tapioca (cassava) decreased risk of developing breast cancer among premenopausal women (P<0.001 and OR=0.35, 95% CI=0.18, 0.65) and a low number of pregnancies (P<0.01), consumption of roots & tubers (P<0.05), usage of chicken meat (P=0.05) increased the risk of breast cancer among post-menopausal women. Further studies seem warranted to confirm the possible protective effect of tapioca consumption. There is an increasing need of breast cancer prevention programs responsive to the cultural practices of the people and the study results should provide leads to cancer control programs especially in rural areas. PMID- 19548368 TI - Interleukin-4-receptor alpha gene polymorphism and the risk of renal cell carcinoma in a South Indian population. AB - The renal cell carcinoma (RCC) is a rare condition, accounting for only 3% of all adult malignancies although constituting 90% of kidney cancers. The tumor is immunogenic and the host immune system may modulate the clinical course of the disease. It has been reported that genetic polymorphisms in the interleukin-4 receptor alpha gene are associated with risk and prognosis in RCCs. The present study is aimed at analyzing the presence and significance of the interleukin-4 receptor alpha Ile50Val and Gln576Arg polymorphisms in a group of RCC patients from South India. PCR-RFLP analysis was performed on genomic DNA isolated from blood samples and the genotypes deduced. A significant association was found between the IL4 R alpha Val/Val genotype and increased risk of RCC (OR: 3.45, CI: 1.15-10.38, P: 0.04). The Val/Val genotype was also found to be significantly associated with increased risk in individuals below 54 years (OR: 5.79, CI: 1.33 25.07 P: 0.03) of age and in females (OR: 7.47, CI: 1.4-39.84, P: 0.03). However, no significant association was observed with the Gln576Arg polymorphism. Stratified analysis based on the genotypes and the stage of tumor revealed no significant association. Thus, the present study indicates that IL4Ralpha could be a candidate gene for assessing the risk of RCC. PMID- 19548369 TI - [Change the procedure or wait for results from large trials?]. PMID- 19548370 TI - [Pulmonary insufficiency after balloon valvuloplasy observed in children]. PMID- 19548371 TI - [Pulmonary insufficiency after tetralogy of Fallot surgery - big clinical problem]. PMID- 19548372 TI - [Systolic and diastolic cardiac insufficiency - are these two different versions of the disease?]. PMID- 19548373 TI - [How to achieve good long term surgical results for coronary artery disease]. PMID- 19548374 TI - [The effect of dronedarone on the frequency of cardiovascular events in patients with atrial fibrillation - ATHENA studies]. PMID- 19548376 TI - [Dronedarone - its place in treatment of atrial fibrillation after the ATHENA study]. PMID- 19548377 TI - [Progress in transcatheter treatment of congenital heart diseases. Report from interventional workshop held in the National Heart Hospital (Sofia, Bulgaria) on 29-30 January 2009]. PMID- 19548375 TI - American Society of Clinical Oncology/College of American Pathologists guideline recommendations for human epidermal growth factor receptor 2 testing in breast cancer. AB - PURPOSE: To develop a guideline to improve the accuracy of human epidermal growth factor receptor 2(HER2) testing in invasive breast cancer and its utility as a predictive marker. METHODS: The American Society of Clinical Oncology and the College of American Pathologists convened an expert panel, which conducted a systematic review of the literature and developed recommendations for optimal HER2 testing performance. The guideline was reviewed by selected experts and approved by the board of directors for both organizations. RESULTS: Approximately 20% of current HER2 testing may be inaccurate. When carefully validated testing is performed, available data do not clearly demonstrate the superiority of either immunohistochemistry(IHC) or in situ hybridization (ISH) as a predictor of benefit from anti-HER2 therapy. RECOMMENDATIONS: The panel recommends that HER2 status should be determined for all invasive breast cancer. A testing algorithm that relies on accurate, reproducible assay performance, including newly available types of brightfield ISH, is proposed. Elements to reliably reduce assay variation (for example, specimen handling, assay exclusion, and reporting criteria) are specified. An algorithm defining positive, equivocal, and negative values for both HER2 protein expression and gene amplification is recommended: a positive HER2 result is IHC staining of 3 + (uniform, intense membrane staining of 30% of invasive tumor cells), a fluorescent in situ hybridization (FISH) result of more than six HER2 gene copies per nucleus or a FISH ratio (HER2 gene signals to chromosome 17 signals) of more than 2.2; a negative result is an IHC staining of 0 or 1 +, a FISH result of less than 4.0 HER2 gene copies per nucleus, or FISH ratio of less than 1.8. Equivocal results require additional action for final determination. It is recommended that to perform HER2 testing, laboratories show 95% concordance with another validated test for positive and negative assay values. The panel strongly recommends validation of laboratory assay or modifications, use of standardized operating procedures, and compliance with new testing criteria to be monitored with the use of stringent laboratory accreditation standards, proficiency testing, and competency assessment. The panel recommends that HER2 testing be done in a CAP-accredited laboratory or in a laboratory that meets the accreditation and proficiency testing requirements set out by this document. PMID- 19548378 TI - [Treatment of aneurysms of the infrapopliteal arteries. Case report and literature review]. PMID- 19548379 TI - Initial anatomic geographic presentation of glottal dysplasia. AB - Glottal dysplasia is likely the most common laryngeal disease with a discernible lesion; however, investigations describing its initial anatomic geographic presentation are rare. To examine this, we identified 52 patients who did not have significant prior treatment or glottal cancer. Thirty-one patients had bilateral disease, so there were 83 vocal folds with precancerous dysplasia. The phonatory mucosa was the dominant disease site in all; the epicenter was on the superior surface in 65 of the 83 folds and on the medial surface in 18 of the 83 folds. The arytenoid mucosa was involved in 8 of the 83 folds. Nineteen of the 52 patients had direct anterior-commissure involvement, and none had interarytenoid mucosal disease. The investigation established the commonly held principle that glottal dysplasia occurs primarily on phonatory mucosa. Given the frequent occurrence and recurrence of glottal dysplasia, treatment goals should focus on disease control to prevent malignant degeneration while preserving the subepithelial superficial lamina propria, necessary for phonatory mucosal pliability, vocal fold vibration, and optimal vocal function. PMID- 19548380 TI - Prevalence of vitamin D deficiency among patients attending a general otolaryngology clinic in South Auckland. AB - OBJECTIVES: We performed a prospective observational study to estimate the prevalence of vitamin D deficiency in patients attending a general otolaryngology clinic in South Auckland, New Zealand. METHODS: From July 21,2008, to August 7, 2008, all new patients presenting to otolaryngology clinics at which one of the authors (D.R.) was present had their vitamin D status assessed by measurement of their plasma 25-hydroxyvitamin D [25(OH)D] level. RESULTS: Of 48 patients, 2% had 25(OH)D levels of 17.5 nmol/L or less (a level associated with osteomalacia), 58% had 25(OH)D levels of 50 nmol/L or less (a level associated with vitamin D deficiency), and 100% had 25(OH)D levels of 80 nmol/L or less. CONCLUSIONS: Most of the patients attending a general otolaryngology clinic in South Auckland are vitamin D-deficient. It is unclear whether low vitamin D levels are associated more directly with otolaryngological disorders or skin type, because the small size of this study and the broad range of conditions seen precluded a meaningful statistical analysis. Further research into the relationship of vitamin D to specific otolaryngological presentations is required. PMID- 19548381 TI - Groningen dilatation tracheoscope in treatment of moderate subglottic and tracheal stenosis. AB - OBJECTIVES: We describe our experience with the Groningen Dilatation Tracheoscope (Karl Storz GmbH & Co, Tuttlingen, Germany) in treating benign subglottic and tracheal stenosis. METHODS: We performed a retrospective survey from a medical record analysis of 26 patients with different origins of benign, grade II (Myer Cotton) subglottic or tracheal stenosis. All patients underwent dilatation tracheoscopy for a total of 64 times (average, 2.46 operations per patient). Five patients had to undergo subsequent operations. Patient data (sex, age at intervention, underlying and other systemic diseases) and operation data (course, intraoperative and postoperative adverse events) were recorded. Analysis of the measured preoperative and postoperative peak flow values was performed. RESULTS: The average predilatation and postdilatation peak flow values were 225 L/min and 331 L/min, respectively. Eighty percent of the patients (21 of 26) were successfully treated with 1 or more dilatation tracheoscopy interventions without supplementary treatment for the stenosis. There is only 1 tracheal cannula dependent patient in the examined population, making the overall success rate of our treatment 96%. No major intraoperative or postoperative complications were recorded. CONCLUSIONS: Dilatation tracheoscopy is a simple, relatively safe, and effective method in the treatment of benign subglottic or tracheal stenosis of various origins. The intervention is minimally invasive and is easily repeatable in case of restenosis. PMID- 19548382 TI - Three-year postimplantation auditory outcomes in children with sequential bilateral cochlear implantation. AB - OBJECTIVES: We report on the auditory abilities and speech performance in quiet and noise of 35 children with sequential bilateral cochlear implantation after 3 years of bilateral implant use. METHODS: Testing was done in bilateral and both unilateral listening conditions. The assessments took place before the second implantation and at several time intervals after fitting. As different auditory tests were used, the children were categorized by their age at the second implantation: younger or older than 6 years. RESULTS: The pure tone averages for the bilateral condition were significantly better than those for either unilateral condition after 12 months of bilateral implant use and remained so from that test interval onward. The speech recognition outcomes in quiet and noise also improved significantly for almost all children after 36 months, although a linear regression analysis showed a beneficial effect of younger age at first implantation on the speech-in-noise results. CONCLUSIONS: Bilateral cochlear implantation offered advantages to all children in comparison with the first implant--even the children who received the second implant after the age of 6 years. Compared to the younger children, the older children needed a longer adjustment period to gain bilateral benefit. However, they obtained similar results after 2 years of bilateral implant use. PMID- 19548383 TI - Acquired atresia of the external auditory canal: recurrence and long-term results. AB - OBJECTIVES: I describe the clinical symptoms and signs of acquired atresia of the external auditory canal (EAC) and the technique used to manage it. METHODS: Forty one consecutive patients affected by acquired atresia of the EAC were assessed by otoscopy, pure tone audiometry, computed tomography, and traditional and echo planar diffusion-weighted magnetic resonance imaging. The anatomic and functional hearing results were evaluated. RESULTS: At surgery for acquired atresia of the EAC, an EAC cholesteatoma was found in 3 of the 41 patients. Twenty-three of the 41 patients were followed for at least 5 years. Recurrence was seen in 9 of the 25 ears (36%) over the entire period of observation. Twenty-one, 23, and 22 of the patients had a normal or nearly normal contour and size of the ear canal at 6 months, 1 year, and 5 years, respectively. The results were similar for the air bone gap. CONCLUSIONS: The main complication following surgery was recurrence. There was recurrence at 6 months in 4 patients (16%). Recurrence was seen in 12% of the cases at the 5-year follow-up. Similar findings were clear on evaluation of the hearing results. This result demonstrates that the surgical procedure, even when performed correctly, did not afford a stable, long-lasting outcome in a cohort of patients. PMID- 19548384 TI - Self-reported listening habits and enjoyment of music among adult cochlear implant recipients. AB - OBJECTIVES: We sought to assess the associations between self-reported listening habits and enjoyment of music, and the following variables: age at implantation, gender, prelingual versus postlingual deafness, duration of deafness, duration of cochlear implant (CI) use, type of CI, speech coding strategy, and speech perception abilities. METHODS: A questionnaire on listening habits and enjoyment of music before the onset of deafness and after implantation was sent to 85 adult CI recipients who had been using the devices for at least 6 months. RESULTS: Of the 53 responders, 39 (73.6%) listened to music after implantation. Listening to music was not significantly related to age at implantation, gender, duration of deafness, duration of CI use, type of CI device, speech coding strategy, or open set speech perception abilities. The 14 nonlisteners were postlingually deafened. The ratings of enjoyment were the same for 22.6% of patients, improved for 26.4%, and worse for 50.9%. Only 2 of 13 patients who played a musical instrument and 14 of 24 patients who sang before the onset of deafness resumed their musical activities. CONCLUSIONS: Despite the decline in listening habits and in the enjoyment of music after cochlear implantation, most patients do listen to music. The changes in listening habits and enjoyment were not related to the selected background variables. PMID- 19548385 TI - Radiographic analysis of cochlear nerve vascular compression. AB - OBJECTIVES: We analyzed whether radiographically demonstrated anterior inferior cerebellar artery (AICA) vascular compression of the cochleovestibular nerve in asymmetric hearing loss could be correlated to either the symptomatic ear or to cochlear nerve diameter. METHODS: We undertook a retrospective case-control study in which patients were enrolled into a database if audiometry demonstrated asymmetry of 20 dB at one frequency, asymmetry of 10 dB at two frequencies, or a difference of 20% on word recognition scores. If AICA vascular contact was demonstrated on subsequent magnetic resonance imaging of the cerebellopontine angle, patients were included in the study. Patients with vestibular schwannoma or Meniere's disease were excluded. The AICA contact was graded by a blinded neuroradiologist according to criteria proposed by McDermott et al. The cross sectional area of the cochlear nerve was measured. RESULTS: Symptomatic ears could be correlated to a decreased cochlear nerve diameter, but not to the degree of AICA penetration into the internal auditory canal. CONCLUSIONS: AICA vascular compression of the cochleovestibular nerve does not appear to correlate to hearing loss or to cochlear nerve diameter. The finding of decreased cochlear nerve diameter in symptomatic ears implies an alternative mechanism for asymmetric hearing loss. PMID- 19548386 TI - Laryngopharyngeal reflux with sore throat: an ultrastructural study of oropharyngeal epithelium. AB - OBJECTIVES: We performed an electron microscopic ultrastructural study of oropharyngeal epithelium in patients with laryngopharyngeal reflux (LPR) and sore throat to evaluate whether dilatation of intercellular spaces could be traced at this level. METHODS: The study included 20 patients with LPR and sore throat and 5 control subjects. The patients were subjected to upper gastrointestinal tract endoscopy and flexible pharyngolaryngoscopy. Oropharyngeal biopsy specimens were taken from the patients and controls for ultrastructural study by transmission electron microscopy. RESULTS: The entire group of patients with LPR showed dilatation of intercellular spaces essentially at the squamous basal and suprabasal levels in their oropharyngeal biopsy specimens, whereas none of the control subjects showed such a morphological marker. CONCLUSIONS: Dilatation of intercellular spaces as a morphological marker can be traced in patients with LPR and sore throat at the level of the oropharynx. This contributes to a better understanding of the pathophysiology of LPR. If this finding is confirmed in a large series, it will represent a cost-effective, relatively noninvasive method for diagnosis of LPR. PMID- 19548387 TI - Human papillomavirus-associated oropharyngeal squamous cell carcinomas: primary tumor burden and survival in surgical patients. AB - OBJECTIVES: We sought to determine whether the primary tumor burden in oropharyngeal squamous cell carcinoma is lower in tumors positive for human papillomavirus (HPV) or in tumors with a smoking- or alcohol-related cause. METHODS: We retrospectively reviewed medical records of patients at our institution who had squamous cell carcinoma of the palatine tonsils, base of tongue, soft palate, or pharynx from 1995 through 2006. The patients underwent primary surgical therapy. The main outcome measures were the HPV status of tumors and nodes and the survival rates (categorized by HPV status). RESULTS: Of 102 treated patients, 48 (47.1%) had HPV-positive carcinomas. Primary tumor size was not significantly different between HPV-positive and HPV-negative tumors (median, 2.5 versus 2.0 cm; p = 0.43). Patients with HPV had a higher prevalence of neck nodal metastases (35% versus 11%; p = 0.003) and high-grade lesions (83% versus 64%; p = 0.03). CONCLUSIONS: Primary tumor burden was not associated with HPV status. Patients with HPV-positive oropharyngeal squamous cell carcinomas had a higher prevalence of neck nodal metastases and high-grade lesions. PMID- 19548388 TI - Clinicopathologic features and outcome of the oncocytic variant of papillary thyroid carcinoma. AB - OBJECTIVES: The purpose of this study was to define the clinicopathologic features and outcome of the oncocytic variant of papillary thyroid carcinoma (OVPTC) with a review of the literature. METHODS: Twenty-three patients with OVPTC over a 10-year period were studied. Demographic, clinical, and histopathologic features and outcome data were analyzed retrospectively. RESULTS: Seventeen women and 6 men, ages ranging from 20 to 76 years (95% confidence interval, 43.0 to 54.48), were studied. Cervical lymph node involvement was found in 43.4% of the patients. Most of the recurrences were associated with thyroid masses greater than 2 cm in diameter. Evaluation of the overall survival data by the Kaplan-Meier method revealed that most recurrences took place earlier than 30 months, and the majority of patients (74%) were well, with no evidence of disease, up to 78 months after the last treatment. All of the OVPTC cases presented as nonencapsulated tumors, and 78.2% demonstrated extrathyroid stromal invasion. CONCLUSIONS: OVPTC is a unique variant of papillary thyroid carcinoma that has distinctive clinicopathologic features. Since OVPTC is often associated with local invasion and may involve cervical lymph nodes, it may require more extensive surgery than classic papillary thyroid carcinoma. PMID- 19548389 TI - Audiometric and vestibular features in a second Dutch DFNA20/26 family with a novel mutation in ACTG1. AB - OBJECTIVES: We analyzed the phenotype in a 5-generation DFNA20/26 family with a novel missense mutation in the ACTG1 gene (c.151G>A) and compared the findings to previous reports on DFNA20/26 families. METHODS: Audiometric data were collected from the family members of a Dutch kindred with the novel ACTG1 mutation. Cross sectional and/or longitudinal analyses were performed on pure tone and speech audiometry data of the mutation carriers. Age-related typical audiograms were constructed. Vestibular examination was performed in all mutation carriers. RESULTS: Overall, high-frequency hearing impairment, most prominent at ages over 30 years, was observed with a progression rate of 1.1 to 2.1 dB/y, increasing with frequency. It ultimately resulted in residual hearing. Speech recognition scores remained good at given pure tone average (1, 2, and 4 kHz) levels, but were slightly poorer than those at similar levels in a group of patients with presbycusis. Vestibular examination did not reveal any consistent, statistically significant abnormalities. CONCLUSIONS: The audiometric phenotype of the Dutch DFNA20/26 family with a novel mutation in ACTG1 was largely consistent with previous reports on DFNA20/26. Considerable variations were found in audiogram configurations within the family. This is the first known DFNA20/26 family that has experienced tinnitus. PMID- 19548390 TI - Radiographic and micro-computed tomographic imaging of lipopolysaccharide mediated bone resorption. AB - OBJECTIVES: Chronic otitis media and cholesteatomas cause hearing loss as a result of bony erosion. This bone resorption is known to be more aggressive when cholesteatomas become infected. The most common organism isolated from both diseases is the gram-negative bacterium Pseudomonas aeruginosa. Lipopolysaccharide (LPS), a major virulence factor found in the gram-negative bacterial cell wall, is well known to incite inflammatory bone resorption. The mechanisms underlying this process, however, are poorly understood. In this study, we developed a mouse model of calvarial osteolysis in which resorption was reliably imaged by plain radiography and micro-computed tomography (micro-CT). METHODS: A murine calvarial model was developed to study bone resorption induced by P aeruginosa LPS. Calvariae from wild-type and knockout mice used in this model were imaged by plain radiography and micro-CT. RESULTS: A high degree of correlation between plain radiography and micro-CT was identified (R2 = 0.8554). Furthermore, maximal LPS-induced bone resorption required functioning toll-like receptor (TLR) 2, TLR4, and myeloid differentiation factor 88 (MyD88). CONCLUSIONS: We have developed a successful model of inflammatory osteolysis in which plain radiography can reliably delineate induced bone resorption. In vivo, we have shown that P aeruginosa LPS signals via TLR2, as well as TLR4 through MyD88. PMID- 19548391 TI - You have to help me. PMID- 19548392 TI - The nightmare on TrailBlazer Street. PMID- 19548393 TI - Parental influence and effects of pro-smoking media messages on adolescents in Oklahoma. AB - OBJECTIVES: This study analyzes the important and influential role that parents can play in modifying adolescents' smoking behavior after being exposed to pro smoking messages seen in the media. METHODS: We used data from the Oklahoma Youth Tobacco Survey of 2002. The data is collected by self-administered questionnaire administered to high school students. We grouped exposure into two categories by merging original responses obtained from the survey. We used Multivariate Logistic Regression models to determine any association between exposure to pro smoking messages seen in media and adolescents'smoking status after controlling for age, sex, gender and extent of parental discussion. RESULTS: Adolescents who lived with someone who smokes and whose parents did not discuss the adverse effects of smoking were almost twice as likely to smoke (OR= 2.03, 95% CI 1.4, 3.0) under the influence of smoking seen on TV or in movies, as those not exposed (p = 0.0004). The odds of smoking among adolescents who are exposed "most of the time" while living with someone who smokes and parents who often discuss the dangers of smoking are 1.75 (CI = 0.81, 3.76) times higher compared to those who are never exposed. However the results are not significant (p = 0.16). Similarly, the odds of adolescents smoking are much less (OR = 1.48, 95% CI 1, 2.2) if they are exposed to lower levels of pro-smoking messages and live with someone who smokes even if parents don't discuss the harmful effects of smoking. Adolescents are more likely to smoke depending on the number of close friends who smoke. CONCLUSION: Our results show that parents can play an important role in protecting adolescents from the effects of pro-smoking messages in the media. Adolescents whose parents discuss the dangers of smoking are less likely to smoke even if they live with a smoker and are exposed to media smoking most of the time. PMID- 19548394 TI - Does treatment of hyperlipidemia with statin drugs reduce all-cause mortality? PMID- 19548395 TI - Part three: Narrative knowledge: the role of narrative in the everyday practice of medicine. PMID- 19548396 TI - Reviewing safety goals. PMID- 19548398 TI - "Oh my quivering heart!". AB - Atrial fibrillation is a multifaceted alteration in cardiac rhythm that requires prompt identification and intervention to ensure optimal client outcome. Many different treatment modalities may be used based on the client presentation and duration of the arrhythmia. Thorough nursing assessment and appropriate nursing interventions are imperative to safeguard clients presenting with atrial fibrillation, both in acute and chronic phases of this condition. PMID- 19548399 TI - Journaling unlocks fears in clinical practice. PMID- 19548400 TI - Self directed floating. PMID- 19548401 TI - Vaccination: compliance controversy. PMID- 19548402 TI - The world's her stage. Interview by Drenan McQuillan. PMID- 19548403 TI - An interview with Professor John Hobkirk. PMID- 19548404 TI - Removable dentures with implant support in strategic positions followed for up to 8 years. AB - PURPOSE: The aim of this study was to analyze prosthetic maintenance in partially edentulous patients with removable prostheses supported by teeth and strategic implants. MATERIALS AND METHODS: Sixty patients with removable partial prostheses and combined tooth-implant support were identified within the time period from 1998 to 2006. One group consisted of 42 patients (planned group) with a reduced residual dentition and in need of removable partial dentures (RPDs) or overdentures in the maxilla and/or mandible. They were admitted consecutively for treatment. Due to missing teeth in strategic important positions, one or two implants were placed to improve symmetrical denture support and retention. The majority of residual teeth exhibited an impaired structural integrity and therefore were provided with root copings for denture retention. A few vital teeth were used for telescopic crowns. The anchorage system for the strategic implants was selected accordingly. A second group of 18 patients (repair group) wearing RPDs with the loss of one abutment tooth due to biologic or mechanical failure was identified. These abutment teeth were replaced by 21 implants, and patients continued to wear their original prostheses. The observation time for planned and repair groups was 12 months to 8 years. All patients followed a regular maintenance schedule. Technical or biologic complications with supporting teeth or implants and prosthetic service were registered regularly. RESULTS: Three maxillary implants were lost after loading and three roots with copings had to be removed. Biologic problems included caries and periodontal/peri-implant infection with a significantly higher incidence in the repair group (P < .05). Technical complications with the dentures were rather frequent in both groups, mostly related to the anchorage system (matrices) of root copings and implants. Maintenance and complications were observed more frequently in the first year after delivery of the denture than in the following 3 years (P < .05). No denture had to be remade. CONCLUSIONS: The placement of a few implants allows for maintaining a compromised residual dentition for support of RPDs. The combination of root and implant support facilitates treatment planning and enhances designing the removable denture. It also proves to be a practical rescue method. Technical problems with the anchorage system were frequent, particularly in the first year after delivery of the dentures. PMID- 19548405 TI - Sinus membrane lift using a water balloon followed by bone grafting and implant placement: a 28-case report. AB - PURPOSE: The objective of this study was to assess the efficacy and safety of a minimally invasive sinus lift using an inflatable water balloon followed by bone grafting and implant placement. MATERIALS AND METHODS: A total of 28 patients with a single tooth missing in the posterior maxilla underwent a water balloon sinus lift, followed by bone grafting and implant placement. Baseline bone height was 4.92 +/- 1.24 mm. Implant site preparation employed a pilot drill and osteotomy followed by water balloon elevation. The mean inflated balloon volume was 0.67 +/- 0.17 mL. Bio-Oss was filled under the elevated sinus membrane using a dedicated instrument. Twenty-eight total implants (diameter: 3.8 to 5.0 mm) were placed. Pre- and postoperative panoramic films or computed tomographs (optional) were taken for every case to measure and compare the results of the sinus membrane lift using a water balloon. Postoperative patient reactions including swelling, discoloration, discomfort, hematomas, and disability were recorded. RESULTS: Successful sinus membrane water balloon lifting procedures were performed in 26 cases; two procedures were aborted due to sinus membrane perforation. A total of 26 implants were placed. The mean inflated balloon volume was 0.67 +/- 0.17 mL and radiographic examination showed the mean elevated height by balloon to be 10.9 +/- 2.06 mm. Computed tomography showed the bone graft distributing evenly around implants. Patients were extremely pleased with the results and needed very little medical attention after surgery. The mean follow up was 15.9 +/- 2.94 months. One implant was lost due to infection. CONCLUSION: The use of a water balloon to elevate the sinus membrane is a truly minimally invasive technique and is associated with very little discomfort. This method has encouraging results, is easy to learn, and is associated with low complication rates. PMID- 19548406 TI - The three-dimensional casting distortion of five implant-supported frameworks. AB - The aim of this study was to assess three-dimensional distortion in cast full arch, screw-retained titanium implant frameworks. A conventional commercial laboratory one-piece casting was used implementing the lost-wax technique. Five wax patterns were fabricated on a die-stone cast poured from a plaster impression of a five-implant brass analog. A reflex microscope was used to determine the three-dimensional casting error. Significant differences were found in distortion between wax patterns and castings, which, given the need to keep within 150 microm of misfit for passivity, were larger than the wax frameworks by between 416 and 477 microm. The greatest distortion occurred at the terminal implant abutments and in the vertical dimensions, but the distortion was inconsistent, indicating its three-dimensional nature. It is doubtful whether any conventionally cast titanium framework can be made to the degree of accuracy required to fit passively on its abutments because of the multiple variables inherent in this process. PMID- 19548407 TI - Effect of an adjustable mandibular advancement appliance on sleep bruxism: a crossover sleep laboratory study. AB - PURPOSE: The objective of this experimental study was to assess the efficacy and safety of a reinforced adjustable mandibular advancement appliance (MAA) on sleep bruxism (SB) activity compared to baseline and to a mandibular occlusal splint (MOS) in order to offer an alternative to patients with both tooth grinding and respiratory disorders during sleep. MATERIALS AND METHODS: Twelve subjects (mean age: 26.0 +/- 1.5 years) with frequent SB participated in a short-term (three blocks of 2 weeks each) randomized crossover controlled study. Both brain and muscle activities were quantified based on polygraphic and audio/video recordings made over 5 nights in a sleep laboratory. After habituation and baseline nights, 3 more nights were spent with an MAA in either a slight (25%) or pronounced (75%) mandibular protrusion position or with an MOS (control). Analysis of variance and Friedman and Wilcoxon signed-rank tests were used for statistical analysis. RESULTS: The mean number of SB episodes per hour was reduced by 39% and 47% from baseline with the MAA at a protrusion of 25% and 75%, respectively (P < .04). No difference between the two MAA positions was noted. The MOS slightly reduced the number of SB episodes per hour without reaching statistical significance (34%, P = .07). None of the SB subjects experienced any MAA breakage. CONCLUSION: Short term use of an MAA is associated with a significant reduction in SB motor activity without any appliance breakage. A reinforced MAA design may be an alternative for patients with concomitant tooth grinding and snoring or apnea during sleep. PMID- 19548408 TI - Changes in the outcome of metal-ceramic tooth-supported single crowns and FDPs following the introduction of osseointegrated implant dentistry into a prosthodontic practice. AB - PURPOSE: The aim of this study was to prospectively analyze the effect that the incorporation of osseointegrated implant dentistry had on the outcome of metal ceramic tooth-supported prostheses that had been in situ for 5 to 10 years. MATERIALS AND METHODS: The 10-year estimated (Kaplan-Meier) cumulative survival of metal-ceramic tooth-supported single crowns (TSCs) and metal-ceramic tooth supported fixed dental prostheses (TFDPs) provided for all patients treated at two time periods was determined and statistically compared (log-rank test). Prostheses in group 1 (404 TSCs and 433 TFDP abutments in 189 TFDPs) were cemented between January 1989 and December 1993, with the outcome determined in 1998. Prostheses in group 2 (539 TSCs and 354 TFDP abutments in 142 TFDPs) were cemented between January 1997 and December 2001, with the outcome determined in 2006. A 500% increase in implants restored occurred between the end of group 1 and group 2 time periods. RESULTS: For TSCs, comparison between groups showed a significantly better survival in group 2 than in group 1 for nonvital TSCs (P = .001), nonvital maxillary anterior teeth (P = .003), nonvital maxillary lateral incisors (P = .008), and nonvital premolars (P = .013). Comparison within groups showed nonvital TSCs had a significantly decreased survival compared to vital TSCs in group 1 (P < .001), but not in group 2 (P = .48). Overall, the estimated cumulative 10-year survival of TSCs in group 2 was 94% +/- 3%. For TFDPs, comparison between groups showed a significantly better survival for nonvital abutments in group 2 than in group 1 (P = .049). Comparison within groups showed nonvital TFDP abutments had a significantly decreased survival compared to vital TFDP abutments in group 1 (P = .001), but not in group 2 (P = .377). Overall, group 2's estimated cumulative 10-year survival for all TFDPs was 90% +/- 6% and for three-unit TFDPs was 97% +/- 2%. Teeth in group 2 failed less through fracture and periodontal disease than those in group 1. CONCLUSIONS: The incorporation of osseointegrated implant dentistry has resulted in a significant improvement in the survival of TSCs and TFDP abutments, nonvital and vital teeth having equivalent survivals for TSCs and TFDPs, and a decrease in supporting tooth failure through fracture and periodontal disease. PMID- 19548409 TI - Dental implants in patients with ectodermal dysplasia and tooth agenesis: a critical review of the literature. AB - PURPOSE: The aims of this article are to critique the available literature on dental implants in patients with ectodermal dysplasia (ED) syndrome and tooth agenesis, review the outcomes of implant therapy in these patients, and provide recommendations on the timing of implant placement for these patients. MATERIALS AND METHODS: Searches were performed using Medline, Embase, All EBM Reviews, and Pre-Medline for articles relating to implant patients suffering from ED. Articles unrelated to the topic of dental implants in patients with ED and tooth agenesis, without abstracts, or in languages other than English were excluded. Selected articles were graded according to levels of evidence based upon guidelines set forth by the Agency for Health Care Policy and Research. Articles found to have a level of evidence of IV were excluded from this study. RESULTS: The literature on dental implants in patients with ED and tooth agenesis was found to be scarce. No randomized controlled or case-controlled studies were found. Only 12 articles were found to satisfy all inclusion criteria. CONCLUSION: Implant survival rates vary between 88.5% and 97.6% in patients with ED and between 90% and 100% in patients with tooth agenesis. Implants placed in adolescent ED patients do not have a significant effect on craniofacial growth, while implants placed in ED patients younger than 18 years have a higher risk of failure. PMID- 19548410 TI - Short-term bone level observations associated with platform switching in immediately placed and restored single maxillary implants: a preliminary report. AB - PURPOSE: The aim of this study was to evaluate the short-term bone level response around immediately placed and provisionally restored implants using a platform switching concept. MATERIALS AND METHODS: Twenty-two implants with a platform diameter of 5.5 mm were immediately placed in healthy maxillary sites in 22 patients. Resultant circumimplant spaces were filled with a mixture of bovine bone matrix and collagen. The implants were randomly divided into two equal groups: 11 implants connected with 3.8-mm-diameter abutments (test group) and 11 with 5.5-mm-diameter ones (control). Provisional crowns were adapted and adjusted for nonfunctional immediate placement on each implant and the final crowns were constructed 2 months later. Posttreatment assessments were carried out by an independent trained observer at the time of implant placement (baseline), at definitive prosthesis insertion, and every 6 months thereafter. These assessments included periapical radiographs, pocket probing depths (PPD), bleeding on probing (BOP), and modified Plaque Index (mPII) on both implants and first proximal teeth. An image analysis software application was used to compare the bone crestal heights at the mesial and distal aspects of the implants. RESULTS: The mean follow-up observation period was 25 months and all implants were judged to be successfully osseointegrated. In the test group, radiographic analysis showed an average bone reduction level of 0.30 mm (SD = 0.16 mm). This mean value was statistically significantly different (P < or = .005) from the average reduction in the control group (mean = 1.19 mm, SD = 0.35 mm). No differences between the two groups in PPD, BOP, or mPII were found. CONCLUSION: This preliminary study suggests that immediate single implant restorations in specific maxillary sites with subsequent platform switching may provide peri-implant alveolar bone-level stability. PMID- 19548411 TI - Fracture resistance of the implant-abutment connection in implants with internal hex and internal conical connections under oblique compressive loading: an in vitro study. AB - The objective of this study was to verify if differences in the design of internal hex (IH) and internal conical (IC) connection implant systems influence fracture resistance under oblique compressive forces. Twenty implant-abutment assemblies were utilized: 10 with IH connections and 10 with IC connections. Maximum deformation force for IC implants (90.58 +/- 6.72 kgf) was statistically higher than that for IH implants (83.73 +/- 4.94 kgf) (P = .0182). Fracture force for the IH implants was 79.86 +/- 4.77 kgf. None of the IC implants fractured. The friction-locking mechanics and the solid design of the IC abutments provided greater resistance to deformation and fracture under oblique compressive loading when compared to the IH abutments. PMID- 19548412 TI - Retention of overdenture posts cemented with self-adhesive resin cements. AB - This study investigated the effects of two self-adhesive resin cements on the retention of overdenture anchor posts after 30 days of aging in water. Forty caries-free human canines were randomly assigned to four test groups. Uni-Anchor posts were cemented to specimens in groups A and B with Breeze and Maxcem self adhesive resin cements, respectively. In groups C and D, Fuji glass-ionomer cement and Fleck's zinc phosphate cement were used, respectively. Specimens were stored in distilled water at 37 degrees C for 30 days. Each specimen was loaded in tension in an Instron universal testing machine. The maximum force required to dislodge each post was recorded. Means and standard deviations (SDs) were calculated and data were statistically analyzed with analysis of variance (ANOVA). Means and SDs were 706.5 +/- 204.6 N for Breeze, 585.1 +/- 213.5 N for Maxcem, 449.2 +/- 181.1 N for Fuji, and 330.4 +/- 120.6 N for Fleck's. ANOVA revealed significant differences among the means (P < .0003). Adhesive failure was observed with all groups except group A, in which eight specimens underwent a cohesive fracture of the dentin. Breeze cement (group A) resulted in the highest retention force and most frequent cohesive failure and thus would be expected to clinically perform in a superior manner. PMID- 19548413 TI - Smile arcs of Caucasian and Korean youth. AB - This study was designed to measure and compare the smile arcs (tooth and lip arcs) of young Caucasian and Korean subjects. Two hundred subjects (100 male and 100 female) were selected from Caucasian and Korean students. Class photographs taken with a digital camera showing the subjects with a posed smile were used for this study. Curves were rendered as semitransparent overlays, which were manipulated over the images using Adobe Photoshop to determine the best fit for tooth and lip arcs. There were statistically significant differences due to ethnicity and gender. Mean lip arcs had greater curvature than mean tooth arcs. PMID- 19548414 TI - Computer-aided design and rapid manufacture of an orbital prosthesis. AB - This clinical report presents a novel approach that integrates a new optical digitizing (scanning) technique, a three-dimensional ocular prostheses database, and the Selective Laser Sintering technique to achieve the computer-aided design and manufacture of an orbital prosthesis. An optical-structured light scanner was used to develop a color digital model of the unaffected orbital contour, which was copied and then mirrored to generate the orbital prosthesis contour data. The ocular prostheses database was applied to ascertain the size and position of the eyeball within the orbital prosthesis. Then, a Selective Laser Sintering machine directly manufactured the wax pattern of the definitive orbital prosthesis from the three-dimensional orbital data. This new approach is time and cost-effective and can be considered an alternative to traditional manual techniques of creating facial prostheses. PMID- 19548415 TI - Randomized controlled clinical study on the accuracy of two-stage putty-and-wash impression materials. AB - PURPOSE: The accuracy of dental impression taking is one major factor influencing the fit of crowns and fixed partial dentures. The aim of this study was to determine the accuracy of three-dimensional (3-D) tooth surface and subgingival tooth surface reproduction using three different silicone materials and the two stage putty-and-wash technique. MATERIALS AND METHODS: From 24 probands, three impressions each were taken with Express STD Putty/Wash (3M ESPE), Optosil/Xantopren L (Heraeus Kulzer), and an experimental ultralight body/putty material (3M ESPE) in a randomized order. The preliminary impression was cut according to established procedures for the two-stage putty-and-wash technique. Master casts were manufactured with a standardized procedure and optically digitized. The 3-D accuracy was analyzed with a computer-aided procedure. The Express STD putty-and-wash impressions were used as a reference. Linear models were used for the statistical analysis. RESULTS: Mean deviations of 27.0 microm and -23.6 microm were found for Optosil/Xantopren L and 26.5 microm and -22.6 microm for the experimental material when analyzing 3-D surface reproduction. The tooth surface (buccal/oral) significantly influenced the accuracy of the surface reproduction. Optosil/Xantopren L showed a more complete reproduction of the subgingival tooth surface than either the experimental or reference materials. CONCLUSION: The accuracy of the 3-D tooth surface reproduction as well as the reproduction of the subgingival tooth surface was not favorably influenced when the ultralight wash material was used with established cutting procedures for the preliminary impression. PMID- 19548416 TI - A survey of clinicians: prioritization of dental treatment in leukemia patients prior to chemotherapy. AB - PURPOSE: Chronic oral and dental infections that may remain benign and asymptomatic in healthy individuals can develop into serious and life-threatening conditions when leukemia patients undergo chemotherapy. There is a general consensus that elimination of chronic dental and oral infections prior to chemotherapy reduces the risk of serious problems that may require hospitalization. The limited time available for dental treatment prior to chemotherapy forces clinicians to prioritize. The aim of this study was to determine how hospital-based clinicians who routinely see leukemia patients prioritize dental treatment prior to chemotherapy. MATERIALS AND METHODS: A survey was conducted that targeted the membership of the American Academy of Maxillofacial Prosthetics because a large number of these clinicians had first hand experience in managing leukemia patients. RESULTS: The consensus was that apical radiolucencies and severe periodontitis must be eliminated. Opinions differed regarding whether and how mild periodontal pathology and caries lesions should be treated. CONCLUSION: Most of the surveyed practitioners believed that infectious processes that invade the bone pose the highest risk if left untreated prior to chemotherapy. PMID- 19548417 TI - [Influence of three point mutations in TNF-alpha promoter gene in clinical manifestations and complications of stomach and duodenal ulcer]. AB - The purpose of our study was to investigate whether polymorphisms -238G/A, 308G/A, and -863C/A within the promoter of the TNF-alpha gene are associated with clinical features of gastric and duodenal ulcer disease in a Russian population. DNA samples of 381 unrelated patients with gastric and duodenal ulcer disease and 216 sex- and age-matched healthy controls were used to determine the TNF-alpha gene polymorphisms by PCR-RFLP assay. Logistic regression analysis has revealed significant associations of polymorphism -308G/A with size of ulcerous defect (p=0.03) and intestinal dyspepsia (p=0.05), polymorphism -238G/A with gastric dyspepsia (p=0.04) and reflux-esophagitis (p=0.05), polymorphism -863C/A with perforation of ulcer (p=0.04). The study results highlight impact of the TNF alpha gene polymorphisms on various clinical features in patients with peptic ulcer disease. PMID- 19548418 TI - [The role of genetic features of Helicobacter pylori in pathogenesis of digestive system diseases: from theory to practice]. AB - In this article the comparative analysis of various methods of Helicobacter pylori verification is shown. Also algorithm of optimization of helicobacteriosis diagnostic, developed on the basis of a comparative estimation of results of various diagnostic techniques is presented. According to this algorithm, for increase of accuracy of helicobacteriosis diagnostic it is recommended to use at least two, and it is better three diagnostic methods: the combination of fast urease test or Helic-test with a histologic method (biopsy material from a stomach corpus) or polymerase chain reaction (ureC gene detection) is preferable. For an efficiency estimation of eradication therapy we recommend "Helic-test" as an effective noninvasive method that is especially important for children. PMID- 19548419 TI - [The volume of the liver and its functional status in primary and metastatic malignant lesions]. AB - 84 patients with primary and metastatic malignant liver lesions predominantly at stage T4N0M0 were studied liver volumes and volumes of foci of lesions, also personal normal liver formula. Obtained data was compared with the functional state of the liver, as determined by the scale of hepatotoxicity NIH-NIAD with additional index points of higher levels of ALT, SCHF and total bilirubin in serum. We found that liver increase in most cases is not only a total foci destruction and predominantly blood disorders and lymphokinesis bodies. Even with 50% dissemination of the malignant growth of functional compensation body remains at a sufficient level, which enables extensive cytoreductional intervention and chemotherapy. PMID- 19548420 TI - [Non-invasive method of diagnostics of biliary tract motor function disorders]. AB - Was developed screening test for early diagnosis of various biliary dysfunction. It is based on determining of elektrodermal resistance and temperature of skin in biologically active points. A new method of diagnosis hasn't contraindications, not require the use of expensive or complicated equipment. PMID- 19548421 TI - [Hemostasis in acute ulcerous bleedings from the stomach and duodenum in experiment]. AB - For the first time in experiment on animals ways of surgical treatment of 'sharp ulcer bleedings' stomach and a duodenal gut with application microporous nikelid titanic implants are developed, the morphological picture healing processes in ulcer defect with implanted microporous nicelid-titanic plate is studied. Introduction in 'ulcer defect' microporous Ti-Ni implants, the sated 7,5% a solution capropher, provides a stable hemostasis, reduces risk of relapse of bleedings (p < or = 0,05). Possibility of performance of operations by the developed technique is proved at sharp ulcer bleedings of a stomach and a duodenal gut. PMID- 19548422 TI - [Impulse activity of masticatory elevator and depressor muscles of the lower jaw in rabbits in conditions of hunger, food intake, and satiety]. AB - In chronic experiences on rabbits impulse activity of the chewing muscles elevating and depressing the lower jaw in conditions of hunger, food intake and satiation was studied. In conditions of alimentary deprivation of animals MU of masseter muscle nonperiodically show low amplitude regular impulse activity of tonic type, and MU of mylohyoideus -- burst type phase discharge activity that is reflected in the structure of the temporary organization of impulse activity of the masseter muscle in the form of monomodal distributions of interpulse intervals and mylohyoideus muscle -- in the form of distribution of bimodal type. Food intake causes reorganization of impulse activity of both muscles which represents in the form of bimodal patterns of distributions of interpulse intervals. The high degree of regularity of burst type rhythmic of action potentials of chewing muscles is the objective criterion of result food-intake behavior. PMID- 19548423 TI - [Timing of the surgery in patients with stopped bleeding from duodenal ulcers with the threat of recurrent bleeding]. AB - This article is devoted choosing the optimal time of operation at patients with stopped ulcerative duodenal bleeding. It has been shown that surgical treatment during the first two days at patients with a high probability of recurrence of bleeding leads to statistically significant mortality reduction in compared with patients who were operated upon within the time frame 3-12 days after stopping bleeding. It is concluded that active surgical tactics at patients with a high probability of recurrence of bleeding at the current stage of modern medicine is more justified than waiting tactic. PMID- 19548424 TI - [Characteristics of outpatient rehabilitation of patients with duodenal ulcer complicated by bleeding]. AB - Now current of a stomach ulcer differs special aggression. Treatment of patients with complicated current gastroduodenal ulcer remains the big problem. The method argon plasma coagulations (APC) now finds primary application for a stop extensive [Russian characters: see text] bleedings. At the same time, the important factor of favorable current of the complicated stomach ulcer is the continuity in treatment of patients and carrying out of adequate therapy from the moment of receipt in a surgical hospital and its continuation at an outpatient stage. PMID- 19548425 TI - [Using of adaptive biocontrolling feedbak in the treatment of irritable bowel syndrome]. AB - In original research are compared results of treatment of irritable bowel syndrome (IBS) by standard technique and treatment with to additional application of the control of a biofeedback. The analysis was carried out on dynamics of clinical symptoms in 1 and 6 months after course of treatment. Research showed improvement of results of treatment IBS in the group with additional treatment of a biofeedback. PMID- 19548426 TI - [The role of the congenital duodenum abnormality in development of the upper gastrointestinal tract and the biliary system diseases]. AB - In article clinical observation of the patient of 17 years with developmental anomaly of a duodenum-megabulbus, transferred in the early neonatal period operative treatment concerning congenital impassability of a duodenum in connection with its atresia, and generated after diseases of upper gastrointestinal and biliary tract is presented. PMID- 19548427 TI - [85-th birthday anniversary of academician RAMS, professor Loginov A.S]. PMID- 19548428 TI - [Academician RAMS Anatolii Sergeevich Loginov (1924-2000)--pioneer of laparoscopy in clinic of internal diseases]. PMID- 19548429 TI - [Current progress of integrative medical therapy in treatment of depression]. PMID- 19548430 TI - [Study and thinking on traditional Chinese medicine syndrome typing of depression]. PMID- 19548431 TI - [Integrative medical experimental studies on depression]. PMID- 19548432 TI - [Application of magnetic resonance imaging technique in integrative medical studies on depression]. PMID- 19548434 TI - [Effect of modified xiaoyao powder for improving sleep in patients with psychological stress insomnia]. AB - OBJECTIVE: To subjectively and objectively assess the effect of Jiawei Xiaoyao Powder (JXYP) on sleep in patients with psychological stress insomnia. METHHODS: A randomized controlled study was conducted in 33 patients with psychological stress insomnia. They were assigned to 4 groups, 4 in the TCM group treated with JXYP, 5 in the Western medicine (WM) group treated with Estazolam, 9 in the integrated medicine (IM) group treated with JXYP plus Estazolam, and 10 in the control group treated with placebo. Quality of sleep in patients was assessed subjectively and objectively before treatment and 6 weeks after treatment by Pittsburgh sleep quality index (PSQI), self-rating scale of sleep (SRSS) and polysomnography (PSG), respectively. RESULTS: Subjective assessment on sleep showed that after 6-week treatment, the scores of PSQI and SRSS remarkably reduced in the TCM, IM and control groups (P < 0.05), while the decrease was insignificant in the WM group (P > 0.05), but no significant difference between groups was shown. The objective assessment by PSG showed that no significant change was found after treatment in parameters of total sleep time (TST), sleep time of phase 1 and 2, slow wave phase, rapid-eye-movement (REM) phase, sleep latency, REM sleep latency, also in long waking and short waking times in all group (P > 0.05), but a significant increase of sleep efficacy (P < 0.05) and an increasing trend of TST (P > 0.05) were shown in the IM group, and an increasing trend of both in the TCM group (P > 0.05). CONCLUSION: JXYP, combined with or without Estazolam, can improve the quality of sleep subjectively, and the combination of the two could enhance the efficacy of sleep in patients with psychological stress insomnia. PMID- 19548433 TI - [Analysis on somnipathy related factors in elderly patients with stroke and comparative study on the efficacy of treatment by traditional Chinese medicine and by estazolam]. AB - OBJECTIVE: To analyze the somnipathy related factors in elderly patients with stroke and to compare effects of traditional Chinese medicine and estazolam on sleep and the followed nervous function. METHODS: The somnipathy related factors in 336 elderly patients with stroke, 221 accompanied with somnipathy and 115 with normal sleep, were studied and analyzed. Moreover, the 221 patients with somnipathy were assigned to two groups, 112 in the treated group treated with TCM according to syndrome differentiation, and 109 in the control group treated with estazolam. Changes of scores rated by neurological deficit scale (NDS), sleep dysfunction rating scale (SDRS), Hamilton anxiety rating scale (HAMA), Hamilton depression scale (HAMD), and treatment emergent symptom scale (TESS) were observed before and after treatment. RESULTS: The occurrence of somnipathy in patients with cerebral hemorrhage was significantly higher than in those with cerebral infarction; it was higher in patients with lesion in brain stem, cerebral hemisphere (frontal lobe), or basal ganglion than in those with lesion in other sites; and patients with severe neurological deficit were more liable to having somnipathy. TCM showed a significant effect in improving the insomnia symptoms in patients, with the total effective rate reaching 81.25% (91/112), which was higher than that in the control group, 65.14% (71/109, P < 0.01). Along with the improving of sleep, marked recovery of the nervous function was shown in both groups, with NDS score reduced significantly (P < 0.05). CONCLUSION: The occurrence of somnipathy in stroke patients was closely related with the property and site of lesion, and the neurological deficit degree of patients. Better sleep is surely favorable for recovery of the nervous function, and TCM shows a favorable efficacy on somnipathy. PMID- 19548435 TI - [Clinical effect of electroacupuncture combined with psychologic interference on patient with Internet addiction disorder]. AB - OBJECTIVE: To observe, adopting randomized controlled method, the therapeutic effect of electroacupuncture (EA) combined with psychologic interference in patients with internet addiction disorder (IAD). METHODS: Forty-seven patients with IAD were assigned to two groups treated respectively with psychotherapy alone (A, 23 cases) and EA plus psychotherapy (B, 24 cases). The psychotherapy was conducted by cognition and behavior method, once every 4 days, for 10 times totally. EA was applied at acupoints Baihui, Sishencong, Hegu, Taichong, Neiguan, Sanyinjiao, etc. once every other day, for 20 times. Changes of scoring by IAD self-rating scale (ISS), anxiety self-rating scale (SAS), self-rating depressive scale (SDS), Hamilton depression scale (HAMD), Hamilton anxiety scale (HAMA) and self-rating sub-health scale (SRSHS) before and after treatment were observed. RESULTS: The total effective rate was 91.3% (21/23) in Group B, better than that (59.1%, 13/22) in Group A. By the end of this study, all scores in Group B, except HAMD, were significantly lower than those in Group A respectively, i.e., for IAD, 33.20 +/- 4.53 vs. 44.00 +/- 5.81; for SAS, 30.90 +/- 6.30 vs. 39.60 +/- 5.80; for SDS, 35.38 +/- 4.59 vs. 39.60 +/- 6.33; for HAMA, 7.50 +/- 2.54 vs. 12.70 +/- 3.68; for SRSHS, 39.60 +/- 5.66 vs. 48.40 +/- 6.91, showing statistical significances (P < 0.05). CONCLUSION: Using psychologic interference alone or combined with EA can significantly reduce the ISS score and significantly reduce anxiety and improve self-conscious health status in patients with IAD, but the effect obtained by the combined therapy is better. PMID- 19548436 TI - [Effect of Qihong decoction on rehabilitation of patients after coronary artery bypass]. AB - OBJECTIVE: To evaluate the clinical efficacy of Qihong Decoction (QHD) on rehabilitation of patients after coronary artery bypass grafting (CABG). METHODS: Seventy-two patients after CABG were assigned to two groups by digital table, the 36 in the treatment group were treated with QHD combined with routine treatment, and the 36 in the control group received routine treatment alone. The treatment lasted for two months in all patients and the efficacy was assessed after treatment. RESULTS: After treatment, the left ventricular ejection fraction (LVEF) increased (P < 0.05), 6-min walking test was improved and score of TCM symptoms decreased in both groups, and inter-group comparisons showed no difference in the efficacy on LVEF, but a better efficacy was shown in the treatment group (P < 0.05) on improvement of walking test and decrease of scores. Moreover, by SF-36 scale scoring, significantly higher scores were shown in the treatment group in domains of general health, pain sensation, vitality, role of emotion, mental health, health transition and physiological capacity (P < 0.05). CONCLUSION: QHD can improve the TCM symptoms and heart function of patients after CABG, enhance their quality of life and accelerate the rehabilitation process. PMID- 19548437 TI - [Long-term efficacy of integrative medicine for treatment of systemic lupus erythematosus]. AB - OBJECTIVE: To investigate the long-term efficacy of integrative medicine in treating systemic lupus erythematosus (SLE). METHODS: Adopting randomized grouping method, the SLE patients were assigned to two groups, 74 in each group, the treatment group treated by combined hormonotherapy with Chinese drugs for detoxifying, and the control group treated by hormonotherapy alone. The period of the stable phase and active phase, clinical manifestations, 24-h urinary protein, systemic lupus erythematosus disease activity index (SLEDAI) score and complement 3 (C3), etc. were observed. RESULTS: After 5-year treatment, the clinical manifestations, including arthritis, hydrohymenitis and fever revealed between groups were significantly different (P < 0.05). Comparison between groups in the 5th year of observation showed that the treatment group had a longer time of the stable phase (305.41 +/- 14.32 days); lower SLEDAI scores (8.28 +/- 4.12) and 24 h urinary protein (0.18 +/- 0.01)g, and a higher level of C3 (0.98 +/- 0.35 g/L) than those in the control group (190.22 +/- 15.43 days, 12.79 +/- 5.43 scores, 0.31 +/- 0.06 g and 0.73 +/- 0.34 g/L), respectively, with statistical significances (P < 0.05). Adverse reactions occurred in 15 cases in the treatment group and 62 cases in the control group, also with significant difference between groups (P < 0.05). CONCLUSION: The combined therapy of Chinese and Western medicine can prolong the stable phase time and shorten the lasting period of SLE in patients, which embodies the view of "treating disease before its onset". PMID- 19548438 TI - [Treatment of albuminuria in gestational hypertension puerpera in the severe preeclampeia stage by TCM therapy for stasis-removing and diuresis]. AB - OBJECTIVE: To explore the TCM therapy for puerperal albuminuria (PA) in patients with gestational hypertension syndrome (GHS). METHODS: Seventy-two GHS patients with PA in the severe preeclampsia stage were assigned to the treated group (38 cases) and the control group (34 cases). They were treated, starting from the postpartum second day, with Nifedipine 10 mg three times per day, but to the treated group, Shenkangbao (SKB, a TCM patent drug for stasis resolving and dinresis) was given additionally at the dose of 10 g twice a day, 3 weeks as one therapeutic course for all. Changes of urinary albumin quality and quantity, plasma total protein and albumin, as well as renal function and blood pressure before and after treatment were observed. RESULTS: Significant difference after treatment between the two groups was shown in terms of percentage of cases with positive albuminuria (0.7 +/- 0.8 vs. 1.5 +/- 0.9), 24-h urinary albumin (520 +/- 480 mg vs. 1352 +/- 861 mg), plasma total protein (74.5 +/- 6.3 g/L vs. 67.8 +/- 6.2 g/L), and plasma albumin (39.4 +/- 4.5 g/L vs. 34.6 +/- 4.3 g/L, all P < 0.01); also in urinary albumin negative inversion rate (92.1% vs. 67.6%, P < 0.01). No significant difference of renal function between groups, and between pre- and post-treatment was found (P > 0.05), as for the blood pressure, it showed a significant difference between pre- and post-treatment in both groups (P < 0.01), but with no difference between groups (P > 0.05). CONCLUSION: TCM therapy for stasis-resolving and diuresis with SKB can promote the eliminating of albuminuria in puerpera in the severe preeclampsia stage. PMID- 19548439 TI - [Primary hepatocarcinoma treated by traditional Chinese medicine combined with transcatheter arterial chemoembolization]. AB - OBJECTIVE: To observe the effect of traditional Chinese medicine (TCM) combining transcatheter arterial chemoembolization (TACE) in patients with primary hepatocarcinoma (PHC), and its influence on patients' immunity, quality of life and adverse reaction. METHODS: Sixty-seven patients with mid-advanced stage PHC were assignd to two groups: the 35 patients in Group 1 treated with TCM combined TACE and the 32 in Group 2 treated with TACE alone. TACE with Gemzar (GEM) and Cisplatinum (DDP) were applied once in both groups, and followed by conventional post-operational management as hydration. The TCM used was prescribed according to syndrome differentiation. Peripheral blood T-lymphocyte subsets and natural killer (NK) cells of patients were measured before treatment (as the base line) and at the end of the 1st and 4th week after treatment (W1, W4), patients' quality of life (QOL) was estimated at the sametime by Karnofsky sore (KPS). Moreover, CT or MRI examination was performed at end of the 4th week to evaluate the short-term efficacy of treatment. RESULTS: Short-term efficacy analyses showed that the effective rate was 51.4% (18/35) in Group 1 and 37.5% (12/32) in Group 2, showing insignificant difference between them (P > 0.05). The levels of CD3+, CD4+, CD4+/CD8+, NK cells actionties and KPS score reduced slightly in both groups at W1, with no significant intergroup difference; but at W4, they did show significant differences between groups, and all indices in both groups were significantly different to those of the baseline and at W1 (P < 0.05 or P < 0.01). Aduerse reaction occurred were mainly fever, digestive reaction and lowering of peripheral white blood cell counting, platelet counting, etc. and the incidence of fever in Group 1 was lower than in Group 2. CONCLUSION: TCM treatment combined with TACE can enhance the immunity and QOL of PHC patients, and alleviate the adverse reaction of chemotherapeutic agents. PMID- 19548440 TI - [Establishment of coronary heart disease model of phlegm-stasis cementation syndrome type in mini-swines]. AB - OBJECTIVE: To establish a disease-syndrome conjugated animal model, the mini swine coronary heart disease (CHD) model of phlegm-stasis cementation syndrome type, by high fat diet feeding and coronary artery balloon injury. METHODS: Mini swine were randomly divided into the control group and the model group, 6 in each group. They were fed with common forage and high fat forage respectively for 10 weeks and the coronary left anterior descending branch in the model group was injured by balloon intervention technique after 2-week feeding to establish CHD model. The model establishment and its physiopathological indices were evaluated by examinations on body mass index (BMI), blood levels of lipid, high-sensitivity C-reactive protein (hs-CRP), body surface electrocardiograph (BS-ECG), coronary angiography and pathological indices. RESULTS: BMI, total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL-C), hs-CRP, sigma-ST and N-ST indicated by BS-ECG in the model group were all higher than those in the control group respectively (P < 0.05 or P < 0.01). Coronary angiography showed coronary lumen narrowed with apparent lumen loss, showing a significant difference as compared with the control group (P < 0.01). In EVG staining, the diameters of lumen in the model group was obviously narrow with intima proliferation, also significantly different to those in the control group (P < 0.01). CONCLUSION: Disease-syndrome combined model for coronary heart disease of phlegm-stasis cementation syndrome type in mini-swine could be established by high fat diet feeding with coronary arterial injury. PMID- 19548441 TI - [Effects of Gadol and Ganoderma spores on the adiponectin signal pathway in hypertrophic myocardium of spontaneous hypertensive rats]. AB - OBJECTIVE: To investigate the alteration of the adiponectin signal pathway in hypertrophic myocardium of spontaneous hypertensive rats (SHR) and to observe the effects of Gadol (GD) and Ganoderma spores (GS) on the hemodynamic parameters and the adiponectin signal pathway of SHR. METHODS: SHRs, 8 weeks old, were randomly divided into four groups: the untreated group, and the three treated groups treated with GD, GS, and GD + GS respectively by gastrogavage for 4 weeks. Controlled with 8-week-old WKY rats, the hemodynamic parameters in all rats were recorded through the carotid artery intubation; the serum level of adiponectin was determined with ELISA; the mRNA expressions of adiponectin receptors (AdipoRs) and carnitine palmitoyl transferase (CPT-1) were determined by RT-PCR; and the protein expression of adenosine monophosphate activated protein kinase (AMPK), both phosphorylated and un-phosphorylated, was detected by Western blot. RESULTS: Compared with the WKY rats, the systolic blood pressure (SBP), diastolic blood pressure (DBP) and myocardial hypertrophy index (MHI) in SHR were significantly higher; the serum levels of adiponectin and phosphorylated AMPK, mRNA expressions of AdipoR1 and CPT-1 in SHR heart tissue were lower (P < 0.05). Compared with the SHR, medication of GD and GS, either alone or in combination, could reduce SBP, DBP and MHI significantly (P < 0.01, P < 0.05), and elevate the mRNA expression of CPT-1 (P < 0.05) in heart, but levels of adiponectin, AdipoR1 and phosphorylated AMPK could only be raised by combined use of the two (P < 0.05). CONCLUSIONS: Adiponectin signal transduction pathway alteration presents in the myocardium of SHR, which might be one of the molecular mechanisms that cause hypertrophic metabolic abnormality. GD and GS could improve the hemodynamic index in SHR, and enhance the level of adiponectin and the expression of its related signal transduction molecules. PMID- 19548442 TI - [Effects of huoxue injection on the adherence of human monocytes to endothelial cells and expression of vascular cell adhesion molecules]. AB - OBJECTIVE: To observe the effect of Huoxue Injection (HXI, a Chinese herbal preparation consisted of red sage, chuanxiong, safflower and red peony root) on the expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), and the adherence of monocytes to endothelial cells in cultured human umbilical vein endothelial cells (HUVECs) injured by oxidized low-density lipoprotein (ox-LDL). METHODS: Model of injured cell was established by adding ox-LDL into the culture of HUVECs, and the model cells were intervened with HXI. The adhesive percentage of the model cells to monocytes was determined by protein quantification; mRNA and protein expressions of ICAM-1 and VCAM-1 were determined by RT-PCR and flow cytometry respectively. RESULTS: After HUVEC being treated with ox-LDL for 12 h and 24 h, its adhesion rate to monocytes increased, with the mRNA and protein expressions of ICAM-1 and VCAM-1 in HUVEC enhanced significantly, showing significant differences as compared with those in the normal control (P < 0.05 or P < 0.01). HXI could significantly reverse the above mentioned changes dose-dependently, showing that these parameters in the HXI intervened cells significantly different to those in the untreated model cells respectively. CONCLUSION: HXI could inhibit the adherence of endothelial cells to monocytes by way of down-regulating the endothelial superficial adhesion molecules, so as to display its protection on endothelial cells, which should be helpful for reducing or suppressing the formation of atherosclerosis. PMID- 19548443 TI - [Influence of modified qianjin huanglian pill on pancreas of mice with insulin resistance]. AB - OBJECTIVE: To investigate the influence of modified Qianjin Huanglian Pill (QJHL), a Chinese herbal compound, on pancreas in mice with monosodium L glutamate (MSG) induced insulin resistance (IR) and its molecular mechanism. METHODS: Controlled by rosiglitazone (Ros), the MSG indiced IR mice were treated with QJHL for 28 days. The laboratory indices were examined including fasting serum glucose (FSG), fasting serum insulin (FSI), insulin sensitivity index (ISI), and morphological changes of pancreas, and levels of insulin receptor (InsR), insulin receptor substrate (IRS1/2) and glucose transporter (GLUT2) mRNA expression in pancreas tissue were determined by the reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: As compared with the model group, the level of FSG was lower (P < 0.01) and ISI was higher (P < 0.05) after treatment in the QJHL treated group, with pancreatic islet hyperplasia and hypertrophy ameliorated significantly (P < 0.01). And these changes were similar to those in the Ros treated group (P > 0.05). Moreover, the level of GLUT2 mRNA expression in pancreas of the QJHL group increased significantly (P < 0.01), while it was unchanged in the Ros group. CONCLUSION: QJHL could reduce IR, ameliorate pathological changes of pancreas, which is possibly related with its action on increasing GLUT2 mRNA expression in the pancreas tissue. PMID- 19548444 TI - [Syndrome identification of CCl4 induced liver fibrosis model rats based on syndrome detecting from recipe used]. AB - OBJECTIVE: To further demonstrate the cognition upon the prominent effect of Yiguanjian and Xiayuxue Decoction on the CCI4 induced rat model of chronic liver fibrosis which have been verified in the previous studies. From the viewpoint of detecting TCM syndrome by recipe used, through this cognition the pathological features of the liver injury model manifesting a syndrome of "Gan-yin deficiency with blood stasis obstructing collaterals" were further explored. METHODS: Wistar rats were randomly divided into the normal group, the model group, and the three medicated groups. All rats, except those in the normal group, were made into chronic liver injury model by subcutaneously injecting CCI4 for 12 weeks. Medication for the three medicated groups began from the 9th week after modeling, with oral administering of Yiguanjian (YGJ, a recipe has been verified to be effective for liver injury and fibrosis), Liuwei Dihuang Decoction (LWDH, a recipe with effects similar to YGJ) and Yinchenhao Decoction (YCHD, a recipe functioned differently) respectively for four weeks. Rats were sacrificed at the end of the experiment, changes of hepatic function, liver pathology and hydroxyproline (Hyp) content in the liver tissue were detected, and contents of Afamin and mRNA expression of alpha-smooth muscle actin (alpha-SMA) in the liver tissue were assayed as well with Real-time PCR. RESULTS: As compared with the normal group, the pathological figures of the chronic liver injury and fibrosis and hepatic function deterioration obviously appeared in the model rats, with the liver content of Hyp and alpha-SMA mRNA expression increased, and Afamin mRNA expression decreased significantly. In the YGJ treated group, the hepatic collagen hyperplasia and deteriorated hepatic function alleviated significantly after treatment, with content of Hyp significantly lowered, and mRNA expressions of alpha-SMA and Afamin restored to some extent (P < 0.05); the same effects on mRNA expressions of alpha-SMA and Afamin were shown in the LWDH treated group, also a decreasing trend of Hyp content (0.05 < P < 0.1), and a significant decreasing of alanine transaminase (ALT) activity was found; while in the YCHD treated group these pharmacological effects mentioned above were not observed at all. CONCLUSION: The pharmacological effects of LWDH and YGJ were similar to some degree, which gives support to the cognition that the feature of chronic liver injury model rat induced by CCI4 is attributable to yin-deficiency sydrome. PMID- 19548445 TI - [Activating effect and mechanism of epimedium on endogenous stem cells]. AB - Stem cells are the cells with capacities of self-renovation, multiplication and differentiation. By activating endogenous stem cells to promote regeneration response has provided a new thinking for the treatment of degenerative diseases. The authors found that epimedium flavonoids (EF) can promote the proliferation and migration of adrenocortical stem cells in corticosterone-treated rats (as a model of Shen-yang deficiency); and through gene-chip test it was showed that EF could significantly up-regulate the growth hormone (GH), growth hormone releasing hormone (GHRH) and other growth factors such as insulin-like growth factor binding protein (IGFBP) and nerve growth factor (NGF) in the model rats. In natural aging rats (as model for Shen deficiency), EF could make the gene expression of multiple tissues youthening, and up-regulate the lowered expressions of GH, GHRH, IGFBP and NGF, etc. Further study on the in vitro isolated and cultivated neuro-stem cells proved that EF and its components have direct promoting actions on stem cell proliferation. All the above-mentioned outcomes indicated that the actions of EF and its extracts on stem cells are possibly the cytological basis for their effects on counteracting the suppression of glucocorticoids on hypothalamus-pituitary-adrenal (HPA) axis and retarding aging; also illustrated that TCM could treat diseases by a way of activating endogenous stem cells through mobilizing and elevating hormones and cytokines levels, and bringing the reserved potential of organism into full play. PMID- 19548446 TI - [Treatment of lower extremity diabetic atherosclerotic obliterans with shuxuetong injection]. AB - OBJECTIVE: To observe the effect and safety of Shuxuetong Injection (SXT) in treating patients with type 2 diabetes mellitus accompanying lower extremity atherosclerotic obliterans. METHODS: The selected patients were assigned by randomized digital table to the treated group (180 cases treated with SXT) and the control group (80 cases treated with alprostadil). The clinical efficacy was assessed after 4 weeks of treatment, and the inner diameters and blood flow of the superficial femoral artery, the posterior tibial artery and the foot dorsal artery were measured and compared before and after treatment using Doppler's ultrasonography. RESULTS: The total effective rate in the treated group was 94.4% (170/180), better than that in the control group, 85.0% (68/80, P < 0.01). Ultrasonic examination showed that the inner diameter and the blood flow of all the arteries measured in the treated groups increased after treatment (P < 0.01), with the increase superior to those in the control group (P < 0.05). No adverse reaction was found during the treatment. CONCLUSION: SXT has the effect of activating blood circulation, dissolving stasis and dredging collaterals, it could reduce or eliminate the formation of thrombi, being effective and safe when applied in clinical practice for the treatment of lower extremity diabetic atherosclerotic obliterans. PMID- 19548447 TI - [Effect of ganji recipe combined with Fructus Bruceae oil emulsion intervention on quality of life in patients with advanced primary hepatic cancer]. AB - OBJECTIVE: To observe and compare the quality of life (QOL) and survival time in patients with advanced primary hepatic cancer (PHC) after they have been treated by the combination of ganji recipe and interventional therapy with Fructus Bruceae Oil Emulsion (FBE) or by the trans-hepatic arterial chemical embolization (TACE) adopting Seldinger's technique. METHODS: Seventy-seven patients with advanced PHC were randomly assigned to two groups, 37 patients in the control group treated with TACE alone, and 40 in the treatment group with the combined therapy. One therapeutic course was 4 weeks, and the intervention therapy was repeated generally after an interval of 4-6 weeks according to patients'condition ,the average intervention frequency was 2.3 for one case. The QOL and the survival time in patients were evaluated after treatment. RESULTS: Significant difference was shown in the changes of the Karnofsky score between the two groups (P < 0.05). The scores suggested that the improvements in the treatment group, including patients'physical energy enhancing, symptoms alleviating and overall QOL improving, were superior to those in the control group (P < 0.05). No significant difference of the 3-month survival rate between the two groups was found (P > 0.05), but the 0.5- and 1-year survival rate were significantly superior in the treatment group to those in the control group (67.6% vs. 42.4% and 38.2% vs. 16.1%, respectively). The median survival time in the two groups was 8.9 and 5.3 months respectively. CONCLUSION: Combination of ganji recipe and interventional therapy with FBE in treating PHC could improve patients' QOL, raise the 0.5- and 1-year survival rate, and prolong the survival time, suggesting TCM treatment has an affirmative significance for enhancing the therapeutic efficacy and improving the prognosis in patients. PMID- 19548449 TI - [Looking over and thinking on the current development of traditional Chinese medicine in California, USA]. PMID- 19548448 TI - [Treatment of constipation by intensive acupoints thread embedding combined with local anal operation]. AB - OBJECTIVE: To search for the best therapy for slow transmission constipation (STC) or mixed constipation (MC) by intensive acupoints thread embedding (IATE). METHODS: A total of 310 patients with constipation were assigned to 5 groups using a stratification control method. The 62 patients of STC in the observation group I were treated by IATE at Dachangshu (BL 25), Tianshu (ST 25), Zhongji (CV 3) and Zusanli (ST 36) acupoints; the 62 of MC in the observation group II by IATE at the same acupoints and local anal operation; both the 62 of STC in the control group I and the 62 of MC in the control group 1I were treated by oral administration of Simo Decoction, while the 62 of MC in the control group III were treated by local anal operation alone. Conditions of defecation were recorded every day in the 6-month follow-up period. Gastrointestinal transit test (GITT) was carried out before and after treatment. One follow-up was conducted at the 1st, 3rd and 6th month. RESULTS: In the 124 patients of the two observation groups, 52 (41.9%) were cured after treatment, they could normally discharge stool, 5 (4.0%) showed markedly effective with their constipation alleviated, but having occasional bowel slackened sensation. In the control group I and II, none was cured or effectively treated, while in the control group III, the efficacy in 8 patients were judged as marked effective, but none was cured. CONCLUSION: IATE is a convenient and effective composite therapy. It shows favorable clinical efficacy for STC, and also for MC in case of combined with local anal operation. PMID- 19548450 TI - [Progress in the research on commonly used anti-cancer traditional Chinese medicine capsules combined with chemotherapy on middle-advanced stage lung cancer]. PMID- 19548451 TI - Maternal obesity as a perinatal risk factor. AB - OBJECTIVE: The aim of the study was to estimate the effect of maternal obesity on pregnancy course, delivery and newborn well-being. MATERIAL AND METHODS: Data about women who delivered in PMMH was analyzed and obese pregnant women (pre pregnancy BMI > or =30) were included in the study group; the rest of the mothers constituted the control group. The pregnancy course, labor and delivery, and newborn well-being were taken into consideration. RESULTS: 4648 women were found in our delivery database, among them 208 (4.48%) were classified as obese. In this group, pregnancy-associated hypertension was common, either non-proteinuric one (8.65% vs 2.39%, p=0.001) or preeclampsia (4.81% vs 1.58 %, p<0.05). There were also more cases of gestational diabetes mellitus requiring insulin therapy (9.62% vs 1.48%, p<0.001) and polyhydramniosis (4.81% vs 2.11%, p<0.05) than in case of controls. The mean gestational age at delivery and newborn general health (estimated by Apgar score, mean umbilical cord pH and the incidence of cases with pH < or =7.10) were similar in both groups. The mean birthweight (3266 g vs 3100 g, p<0.05) and the incidence of macrosomia (20.19% vs 5.69%, p<0.001) were significantly higher in the study group. The delivery mode was comparable in both groups, with the marked tendency towards higher incidence of elective cesarean sections in case of obese mothers (27.88% vs 19.90%, p=0.01). CONCLUSION: Maternal obesity is a significant perinatal risk factor; with pregnancy associated hypertension and gestational diabetes requiring insulin therapy in obese mothers and macrosomia in newborns as most common complications. PMID- 19548452 TI - [The assessment of leptin concentration and Vascular Endothelial Growth Factor (VEGF) in relation to the body mass index since 20th week of pregnancy]. AB - High body mass index and obesity in pregnancy signify an increased obstetrical risk. Obesity before pregnancy qualifies a patient into the group that demands more attention from the obstetrician. Leptin and VEGF are among numerous factors that influence the pregnancy course and outcome. MATERIAL AND METHOD: The study was conducted in a group of pregnant women from 20-24 weeks of gestation in High Risk Pregnancy Clinic, Medical University Lodz, between 2005-2007. The study group consisted of 30 pregnant women with BMI > or =30 and the control group consisted of 25 pregnant women at the same gestational age and BMI < or =25. Concentrations of leptin and VEGF were measured in venous blood every 4 weeks. RESULTS: More body mass gain during pregnancy was observed in the group of women with BMI > or =30 when compared to the group of BMI < or =25. Mean value of leptin was higher in the group of obese women. No difference was found in leptin concentration measured every 4 weeks. The correlation between leptin concentration and BMI was found in the group of obese women. The concentration of VEGF was higher in controls than in the group of obese women. The mean concentration of VEGF measured every 4 weeks in both groups was similar. The highest values of VEGF were found in 20-24 and 30-34 weeks of pregnancy in women with normal BMI. CONCLUSIONS: 1. The synthesis of leptin depends on body mass, not the duration of pregnancy. 2. Obesity in pregnancy is connected with decreased VEGF synthesis. PMID- 19548453 TI - [Personality profiles of pregnant and postpartum women with symptoms of perinatal depression: the differences of self image in the sphere of psychological needs]. AB - OBJECTIVES: The aim of the following study was to assess personality profiles of pregnant and postpartum women with symptoms of perinatal depression and to estimate the differences in their real and ideal self image in the sphere of psychological needs. MATERIAL AND METHODS: 50 pregnant (between 32-40 weeks of gestation) and 26 postpartum women (2-5 days after the delivery) were recruited for the study. Edinburgh Postnatal Depression Scale (EPDS) was used as a screening test for symptoms of pre- and postnatal depression. Personality profiles were evaluated by means of the Adjective Check List (ACL) with 15 need scales. RESULTS: Postpartum women in comparison to pregnant patients described their self image at a statistically significant higher level as far as the needs for domination, nurturance, affiliation, heterosexuality and exhibition were concerned. No significant differences were observed in these groups of patients in the domain of ideal self image. CONCLUSIONS: 1/ pregnant and postpartum women, suffering from perinatal depressive disturbances, differ in terms of the self assessment of some of their real psychological needs, but do not differ in terms of the ideal self image; 2/ observed differences between pregnant and postpartum women are basically consistent with the change of the role which is associated with the pregnancy and postpartum period; 3/ a positive change of the real self image, which is connected with the maternal role in the postpartum period, takes place in women irrespectively of their depression; 4/ observed multiple and significant differences between the ideal and real self image in pregnant as well as postpartum women with perinatal depressive disturbances indicate their lack of self-acceptance which requires individualized psychological intervention. PMID- 19548454 TI - [Detection of selected mutations in the CFTR gene in single cells for the use in preimplantation genetic diagnosis of cystic fibrosis]. AB - Cysitic fibrosis (CF) is one of the most common autosomal recessive diseases. The median life expectancy is currently about 30 years. Because of a considerable social meaning of CF, it is very important to create more specific and efficient methods diagnosing defects in the gene CFTR that are responsible for CF. The use of these methods in preimplantation diagnosis could prevent the disease and also the carrier state. OBJECTIVES: The aim of this study was to construct a diagnostic test for five CFTR gene mutations, the most frequent in Polish population, which can be used to analyze single cells while preimplantation genetic diagnosis. MATERIAL: The material used in research were 60 single cells- blastomers. The positive controls of the CFTR gene mutations: delF508, R117H, G542X, R553X, dele2,3 were obtained from blood of patients with CF. At first Nested Multiplex PCR reaction was performed. Its products were used as DNA template for the next reaction--ASA (allele-specific amplification) PCR. SSCP method (Single Strand Conformation Polymorphism) was used as a verification method. RESULTS: The diagnostic test for the presence of the mutations: delF508, R117H, G542X, R553X and dele2,3 (when homozygosity) was constructed. Mutations: delF508 (3 cells--5%), G542X (2 cells--3.33%) and R553X (1 cell-- 1,.7%) were detected as a result of performing ASA PCR analysis for the presence of the CFTR gene mutations in 60 blastomers. The SSCP analysis for the mutations delF508, R 17H, G542X, R553X confirmed the results of ASA PCR analysis. CONCLUSIONS: The constructed diagnostic test for the mutations delF508, G542X, R553X, R 17H and dele2,3 can be used to analyze single cells during preimplantation genetic diagnosis. PMID- 19548455 TI - [Polymorphism of osteoprotegerin gene and osteoporosis in postmenopausal women]. AB - INTRODUCTION: Osteoprotegerin (OPG) plays a crucial role in the control of bone resorption through competitive inhibition of receptor activator of nuclear factor kappaB ligand (RANKL). This process leads to inhibition of osteoclasts differentiation and activity. The aim of the following study was to evaluate the distribution of genotypes of -163A>G and 1181G>C polymorphisms in OPG gene and analyze their relationship with bone mineral density (BMD) and other parameters of bone turnover in population of Polish postmenopausal women. MATERIAL AND METHOD: The study included 310 postmenopausal Caucasian women (54.48+/-8.53 years); 139 women with osteoporosis, 107 with osteopenia and 64 healthy women. Genetic analysis was performed by PCR/RFLP reaction. BMD value was measured by dual energy X-ray absorptiometry (DXA). RESULTS: For -163A>G polymorphism the higher frequency of heterozygotes AG (26.6 vs. 18.7%, ns) and slight overrepresentation of mutated G allel (14.1 vs. 10.9%, ns) in the osteoporosis group was observed. The frequency of recessive CC homozygotes and C alleles of 1187G>C polymorphism did not differ among the investigated groups. The distribution of particular haplotypes of -163A>G and 1181G>C polymorphisms in all subgroups was similar. Correlation between values of investigated parameters of bone turnover and frequency of genotypes of investigated polymorphisms has not been observed. CONCLUSIONS: The overrepresentation of heterozygous AG genotype and mutated G allele of -163A>G polymorphism of OPG gene in the group of women with osteoporosis might suggest the significance of this variant in the development of osteoporosis. A more extensive analysis of genetic variants of RANKL/RANK/OPG signal pathways, joint with an investigation of modulated influence of estrogens, TNF-alpha or several interleukin influencing the development of osteoporosis is necessary. PMID- 19548456 TI - [Usefulness of the SCC, CEA, CYFRA 21.1, and CRP markers for the diagnosis and monitoring of cervical squamous cell carcinoma]. AB - OBJECTIVE: To determine the usefulness of the SCC, CEA, CYFRA 21.1, and CRP markers for the diagnosis and early monitoring after treatment completion in women diagnosed with cervical squamous cell carcinoma. MATERIAL AND METHODS: Serum of 140 patients with diagnosed cervical squamous cell carcinoma was investigated. The women with the advanced stage of cervical carcinoma (FIGO IIIB) were divided into two subgroups: with positive and negative outcomes of the treatment. Levels of SCC, CEA, CYFRA 21.1, and CRP were measured before the treatment and immediately after the completion of radiotherapy. Immunochemical methods were used to measure proteins in both serum and plasma samples. RESULTS: 75% of the markers measured were within the reference range for FIGO stage I. The marker levels rose with the clinical progression of the disease. The median levels of all markers and the CRP levels in both groups were compared before the treatment. Only in case of CEA a considerable variation between these groups was observed. Elevated levels of CRP were observed twice more often in patients with negative outcome of the treatment. After the treatment, a significant decrease in all marker levels was observed in patients with positive outcome when compared to the levels at the moment of the diagnosis. CONCLUSIONS: SCC, CEA and CYFRA 21.1 markers show low diagnostic sensitivity in early stages of the disease in women diagnosed with cervical squamous cell carcinoma. The concentration of markers measured before the treatment, particularly CEA, may prove to be of prognostic value for women diagnosed with advanced cervical cancer. Certain markers may prove useful in the assessment of the therapy used. Measuring the CRP before the treatment may aid the prognosis of response to treatment in these patients. PMID- 19548457 TI - [Evaluation of stress responses in parents of newborns hospitalized in the Intensive Care Unit]. AB - AIM: To analyze the results of an investigation about the influence of newborn hospitalization at the Neonatal Intensive Care Unit on emotional status of the parents. MATERIAL AND METHODS: The survey was conducted among 98 parents of the newborns hospitalized in the NICU with the use of the PPS-NICU questionnaire. The questionnaire included potential stress factors such as ward environment, treatment procedures as well as the role of the parents in such circumstances. RESULTS: Factors causing the most severe parental stress proved to be: respiratory distress of the newborn, the necessity of mechanical ventilatory support and vital signs monitor alarm activation. Differences in stress levels between mothers and fathers were presented on the basis of their own estimation of the relationship with the child and understanding of their individual parental roles. CONCLUSIONS: Results of the study indicate that it is possible to decrease the stress level among parents of the NICU patients by explaining the background of the disease, the current clinical condition of the newborn, the necessity of diagnostic and treatment procedures and involving the parents in the basic care of the newborn. PMID- 19548458 TI - [Evaluation and treatment of adolescent girls with hirsutism]. AB - Clinical manifestations of androgen excess which are skin and hair related (hirsutism, acne, alopecia) are common and distressing symptoms for an adolescent girls. During puberty and at the time of the first menstruation cycles, physiological hyperandrogenism can be observed. The causes of hirsutism can be various, including familial, idiopathic, and those, caused by excess androgen secretion by the ovary (PCOS, tumors), or by adrenal glands (congenital adrenal hyperplasia, tumor), or exogenous pharmacologic sources of androgens. The diagnosis and treatment of hirsutism remains quite problematic due to innumerous endocrinologic aspects and unsatisfactory treatment results. Androgen excess during puberty must be appropriately recognized, clinically evaluated and treated. Pharmacologic and cosmetic treatments may have beneficial effect. Oral contraceptives and antiadrogens combinations may be recommended as the treatment of choice in adolescents. PMID- 19548459 TI - [Inflammatory bowel diseases in pregnancy]. AB - Ulcerative colitis (CU) and Crohn's disease (CD) are called inflammatory bowel diseases (IBD). In the light of contemporary knowledge it is believed that a common feature of CU and CD is chronic gut inflammation which is the result of a combination of genetic, immunologic and environmental factors. CU and CD incidence peak is observed in the age group from 20 to 40 and, therefore, women in reproductive age constitute the bulk of it Dagnostics is based on biochemical, endoscopic and histologic examinations whereas treatment is based on long-term anti-inflammatory and immunosuppressive pharmacotherapy. Knowledge of the influence of IBD on pregnancy course and proper treatment allows us to reduce the risk of serious obstetric complications and significant adverse reactions to medication. In this review we discuss the issue of inflammatory bowel diseases affecting pregnant women, their impact on pregnancy and risk of obstetrical complications, as well as pharmacotherapy safety issues. PMID- 19548460 TI - Ruptured ectopic pregnancy mimicking acute pancreatitis. AB - INTRODUCTION: Ectopic pregnancy may lead to massive haemorrhage, infertility or death. Prompt diagnosis and treatment are crucial to save patients who would otherwise die. Serum amylase and lipase measurements are known biochemical markers of pancreatic inflammation and a recognized finding that may help diagnose acute pancreatitis. To the best of our knowledge (Medline, Pubmed, Cochrane Library have been researched) the following study presents the first case of ruptured ectopic pregnancy accompanied by markedly elevated amylase and lipase levels mimicking acute pancreatitis ever reported. CASE REPORT: A previously healthy, nulliparous 35-year-old woman was admitted to hospital with a 2-day history of abdominal pain and vomiting. Her last menstrual period was 7 weeks before presentation. At the admission, the patient was hemodynamically stable. The abdomen was soft with tenderness in its mesogastric area. Blood tests revealed markedly elevated activities of the pancreatic enzymes. Acute pancreatitis was the early clinical diagnosis and subsequent therapy was initiated. After 12 hours the condition of the patient suddenly worsened. She was clinically shocked with pallor, hypotension and tachycardia. Laboratory tests revealed anaemia and increased activities of pancreatic enzymes. An ultrasound examination demonstrated an accumulation of intraperitoneal fluid in the pelvis. Subsequently, the patient was subjected to immediate laparotomy. The peritoneal cavity contained large amount of blood. A cystic mass was found and extracted from the ruptured and bleeding right fallopian tube. Histological examination confirmed a rupture of an ectopic pregnancy of a 6-week-old foetus with an intact gestational sac. The patient made an uneventful recovery and was discharged from hospital after 8 days. CONCLUSIONS: Our case proves that a misdiagnosed ruptured ectopic pregnancy in the event of elevated activities of pancreatic enzymes may lead to delayed diagnosis of haemorrhage to peritoneum, resulting in hemodynamic instability. PMID- 19548461 TI - Hydrothorax treated in utero and monitored by fetal echocardiography. AB - Primary fetal hydrothorax is a relatively rare disorder, occurring in 1 out of 15,000 pregnancies, but connected with high perinatal mortality. In the event of little or no progress of the effusion or stable clinical course, conservative management seems to be the most appropriate course of action. In nearly half of the cases, a placement of only one pleuroamniotic shunt enables a total regression of the primary fetal hydrothorax and, consequently, a favourable outcome. The remaining cases require the placement of yet another shunt. In the case of repeated shunt failure, the prognosis is very poor. In the following paper we have presented a fetus with a giant re-accumulating hydrothorax after thoracocentesis, treated by pleuroamniotic shunts and monitored by fetal echocardiography and cardiovascular profile score. Despite dramatic fetal presentation and rapid re-accumulation of the hydrothorax, monitoring by fetal echocardiography and cardiovascular profile score proved the repeated fetal needling and double shunting to be safe. If there are no complications such as premature rupture of the membranes and preterm delivery, fetal echocardiography is a sufficient way of monitoring fetal well-being and the results of intrauterine procedures. PMID- 19548462 TI - [Polish Gynaecological Society guideline on prenatal diagnosis]. AB - Prenatal diagnosis is a multidisciplinary issue where obstetricians, geneticists, neonatologists and doctors representing other specialities are involved. The guideline will provide up-to-date information, based on clinical evidence optimal techniques and timing, training and competence and clinical governance issues. Prenatal screening for chromosomal defects should be performed in concordance with Polish Gynaecological Society guidelines and recommendations on antenatal care, ultrasound in pregnancy and fetal therapy, and Fetal Medicine Foundation (London, UK) rules. There is no doubt that maternal age alone as a method of screening for chromosomal abnormalities should be abandoned as it has low Detection Rate with high False Positive Rate hence high Invasive Procedure Rate and unnecessary high pregnancy loss rate. The Working Party recommends that screening methods based on ultrasound examination at 11(+0)-13(+6) wks and maternal serum biochemistry should be implemented. Special attention must be paid to ensure that sufficiently high Detection Rate is achieved (at least 75% for 5% False Positive Rate) in screening for trisomy 21. PMID- 19548463 TI - [The life-style of the industrial enterprise workers]. AB - To develop the theoretically substantiated recommendations on the formation of healthy life-style and decrease of morbidity among the enterprise workers the public opinion poll on the sampling of 955 respondents was organized. The specially developed questionnaire was applied. The study was carried out in 2007 in the Republic of Dagestan, the city of Makhachkala, on the industrial enterprise "The Gadjiev Plant". The study revealed that among the respondents the stated average monthly income per family member accounted for 1200 rubles in 20%, up to 3000 rubles in 48%, from 3000 to 5000 in 23% and more than 5000 rubles in 9.35%. It is established that in 67% of respondents more than a half of family budget is spend on food stuff. More than 70% of workers drink alcohol, and 33% smoke tobacco. In the structure of causes of unfavorable family relationships first position is for material non-security, second position is for housing non security and third position is for conjoint residence with parents. The study results permitted to develop the target program on health improvement of working and mode of life conditions, formation of healthy life-style of the industrial enterprise workers. PMID- 19548464 TI - [The medical science of the twentieth century: preliminary outcomes]. AB - The major events, problems and achievements of medicine of twentieth--early twentieth first centuries are indicated. The main directions of implementation of the strategy of population health promotion in Russia are identified. The practical activities in caring out the tactic tasks of public health system and medical science development are outlined. The significance of resolving the health problems for successful implementation of the concept of public health development up to the year of 2020 is demonstrated. PMID- 19548465 TI - [The means to enhance the quality and effectiveness of work of local physicians and general practitioners]. AB - The content and labor of organization of 27 local physicians and 19 general practioners was studied within the time period of 355 days (2307 hours) using the photo time-sheet technique. The study was held from 2005 to 2006 in three cities and five rural districts of the Republic of Dagestan. It is revealed that the shortcomings leading to the shortage in finding time for implementation of basic professional functions are common in labor organization of local physicians and general practitioners. To the curative diagnostic and preventive activities the general practioners allocate only 40.5% of working hours and general practitioners even less - 35.5%. With that the time cost of formalization of medical documentation is too overwhelming (31.1-38.2%) and other time spending is considerable as well. In the structure of population visiting the general practioners and local physicians according the classes of diseases first place is for the diseases of respiratory system (29 and 35.6% respectively), second place is for the diseases of circulatory system (18 and 20.6%) and third place is for the diseases of musculoskeletal system (7.7 and 9.8%). On basis of research data the guidelines and proposals are developed concerning the enhancement of quality and effectiveness of professional activities of local physicians and general practitioners. PMID- 19548467 TI - [The analysis of the structure of nurse personnel work time in the rehabilitation treatment department]. AB - The workload of medical nurse is increased significantly due to such actual organizational technical conditions of work as the implementation of hi-tech technologies and outfits in the rehabilitation departments, the increase of proportion of patients with painful course of illness, the wide application of combination methods of rehabilitation treatment. The modern approaches to the organization of nursing process call for broadened standardization of all its aspects. The investigation of the structure of basic work time of nurse personnel the principle of differentiation of working operation on depended, interdependent and independent nurse interventions is to be used. Actually, only small part of the nurse depended rehabilitation interventions such as the volume of medical procedure supply is allowed for. The working hours efforts to the implementation of interdependent and independent nurse interventions are not considered. The attention should be paid to the alteration of the structure of auxiliary work time and the new system of circulation of documents in the inpatient department. In this connection the need for a revision of available standards with a glance to the new demands and changed conditions of medical care rendering. PMID- 19548466 TI - [The sociological evaluation of quality of medical care rendered to the patients with body overweight and obesity]. AB - The evaluation of quality of medical care rendered to the patients with body overweight and obesity was carried out in the conditions of curative preventive institution and private medical clinics on the municipal level. The study revealed the problems related to the organization of medical care provision to this category of patients addressing to the public medical institutions. The conclusion is made about the need of enhancing the actual system of medical care of patients with body overweight and obesity. It is rational to consider patients' opinion during the optimization of the available high quality medical care. PMID- 19548468 TI - [The standard support of preventive examinations of working population]. AB - The article presents the analysis of the standard support of implementing the first stage of the program of the working population additional dispanserization in the framework of endocrinologist activities. PMID- 19548469 TI - [The organization of the comprehensive prevention of urolithiasis among ferrous metallurgy workers]. AB - The purpose of study is to evaluate the effectiveness of the set of preventive measures as applied to 321 workers of basic ferrous metallurgy specialties (steel makers, mill men, hot metal shearers). During the clinical examination all the workers were divided on three groups: the workers without any pathology (11.83%, the first group), the workers with metabolic disorders only without urolitiasis (64.81%, the second group) and the workers with urolitiasis diagnosis approved by ultrasonography (23.36%, the third group). The effectiveness of rehabilitation measures was evaluated during half a year (diet therapy, drinking regimen, medicinal plants treatment). After the course of preventive measures was applied the overall health condition of most workers ameliorated and the number of workers with urolitiasis development risk factors reliably decreased up to 6-12%. PMID- 19548470 TI - [The social hygienic characteristics of educational personnel in vocational medical schools]. AB - The article presents the analysis of the structure of educational personnel in the system of vocational medical schools in Orenburgskaya Oblast. The need in the target and advanced training of teachers in the medical colleges and vocational medical schools is emphasized. PMID- 19548471 TI - [The characteristics of health care services of officials in the Republic of Kazakhstan]. AB - The organizational and functional structure of health care services of officials in Kazakhstan is analyzed. It is demonstrated that the strategic ultimate target to ameliorate health conditions of this representative social group is achieving through the multilevel systematization of health impacting risk factors, medical support characteristics and strategic management qualities. PMID- 19548472 TI - [The characteristics of morbidity in women of fertile age residing the area of Tadjikistan aluminum factory]. AB - The social hygienic characteristics of deteriorated living environment in Surkhan Daria area around the Tadjikistan aluminum factory are described. The harmful chemical substances negatively impacting local population health especially women of fertile age are indicated. The structure of morbidity of women of fertile age based on the data of overall medical examination is analyzed. It is demonstrated that the most frequent pathology in the examined area are diseases of endocrine system, blood and hematopoietic organs, digestive system and genitourinary system. The results of studying the poor health conditions developed under the impact of poor ecological environment due to long-term and harmful influence of waste substances of factory on the air, soil and food are discussed. The system of measures targeted on the improvement of the medical care of women of fertile age residing the aluminum factory territories are proposed. PMID- 19548473 TI - [The prevalence of brain malignant neoplasms in the Republic of Kazakhstan]. AB - The article presents on the data related to morbidity of brain malignant neoplasms in the Republic of Kazakhstan as a whole and its regions. The increase of morbidity during the period from 1996 to 2005 more than 1.8 times was revealed. The higher levels of morbidity (from 4.2 to 5.8 cases on 100 000 of population) were revealed in Kzylordinskaya, Pavlodarskaya, Vostochno kazakhstanskaya oblasts and in the city of Almaty. The morbidity of this pathology is higher in males in comparison with females (ratio 1.2:1.0) and these differences are most pronounced in Vostochno-kazakhstanskaya and Pavlodarskaya oblasts. PMID- 19548474 TI - [The becoming and development of sanitary epidemiologic service in the Russian Federation]. PMID- 19548475 TI - [The medical activities of the Imperial Philanthropic Society (1804-1816)]. PMID- 19548476 TI - [G. A. Kolosov and his input into history of medicine]. PMID- 19548477 TI - First things first. PMID- 19548478 TI - Learning the skills of conflict engagement. PMID- 19548479 TI - Hastings nurse-midwives: practicing evidence based care. Interview by Heather Swanson and Kymberli Shrout. PMID- 19548481 TI - Evidence-based strategies to address nurse manager engagement. PMID- 19548480 TI - Motivation and decision making. PMID- 19548482 TI - It's never too late for education. PMID- 19548483 TI - Rising to the legislative challenge. PMID- 19548484 TI - The low bid gets the patient. PMID- 19548485 TI - Dentistry in New York State faces uncertain future. PMID- 19548486 TI - NYSDA changes peer review protocol. PMID- 19548487 TI - Why are my colleagues reviewing my malpractice claim? PMID- 19548488 TI - The effect of frequent clinical use of dental unit waterlines on contamination. AB - Three dental units with self-contained water systems in an outpatient teaching dental clinic were treated with a proprietary chlorine dioxide waterline cleaner. Three similar units were used as controls. After four weeks, test and control units were crossed over. Water samples were taken from each line on each unit and from the sink faucets at six time periods; and the frequency of use of each line was recorded. Statistical analysis showed that increased frequency of use of waterlines did not affect lines that were chemically treated, but was associated with less contamination of untreated lines. PMID- 19548489 TI - Unusual case of gouty tophous within oral-maxillofacial region. Case report. AB - Manifestations of systemic diseases can appear in the oral-maxillofacial area. They can be microbial, hematologic, endocrine, metabolic, genetic, nutritional, granulomatous and immunologic, and are based upon the disease. We present an extremely rare case in which deposits of crystallized monosodium urate appeared in the right paranasal region. PMID- 19548490 TI - In vitro evaluation of apical extrusion of bacteria following use of new rotary instrumentation system. AB - The aim of this study was to evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using two engine-driven nickel-titanium instruments (Flex Master and V-Taper). Seventy extracted maxillary central incisor teeth were used. After preparing access cavities, root canals were contaminated with a suspension of Enterococcus faecalis, then dried. The contaminated roots were divided into two experimental groups of 30 teeth each and one control group of 10 teeth. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 hours. Colonies of bacteria were counted, and the results were given as number of colony-forming units. The data obtained were analyzed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with alpha = 0.05 as the level for statistical significance. Results showed that there was no significant difference as to the number of extruded bacteria between the two engine-driven systems (P > 0.05). Both engine-driven nickel-titanium systems extruded bacteria through the apical foramen. PMID- 19548491 TI - Desmoplastic fibroma of the anterior mandible. Case report and review of literature. AB - Desmoplastic fibroma is a rare and locally aggressive lesion that affects the long bone and the jaw bones. Mandibular bone involvement has been mostly reported in the posterior segment; anterior region presentation is rare. This case report highlights the diagnosis, management and treatment of a midline mandibular involvement. It includes a literature review. PMID- 19548492 TI - Collaborative approach to dental care provision in resource poor Honduran community. AB - Access to dental care is becoming an increasing problem in the United States, as in other parts of the world. The dental program at the University of Rochester clinic in Honduras, as well as other projects, have demonstrated that the ability to deliver quality care can be enhanced by expanded use of medical personnel, other than dentists, and even by civic action. While there is no substitute for the trained dentist, perhaps techniques learned in these projects can alleviate some of the predicaments until such a dentist is accessible. PMID- 19548493 TI - Lower lip sialolith. AB - The authors report a sialolith of the lower lip that was asymptomatic and radiolucent. Its presence was only detected by the patient when she felt a hard nodule with her tongue. PMID- 19548494 TI - Sturge-Weber syndrome: physician's dream; surgeon's enigma. AB - Sturge-Weber syndrome (SWS) is a neurocutaneous syndrome consisting of port wine stain (facial nevus flammeus), congenital glaucoma and underlying anomalous leptomeningeal venous plexus with lack of normal cortical venous drainage. Neuromuscular, ocular, dermatologic and oral manifestations are variously present with this syndrome. The syndrome presents with various oral findings and the diagnosis of these cases is based primarily on clinical findings. Considering the complications that may be encountered during the treatment and the patient's own perception of the condition, it is imperative that the diagnostician and surgeon have thorough knowledge of the condition. Here we report a case of SWS with facial angiomas, seizures and intracranial calcifications. PMID- 19548495 TI - Restoration of endodontically compromised anterior teeth with fiber posts and zirconia all-ceramic system. Case study. AB - Deciding what material to use to restore endodontically treated teeth complicated by total or partial loss of coronal tooth structure is sometimes difficult for the clinician. It is important to use materials that minimize risk and best accommodate clinical conditions and patient desires. This report describes the restoration of endodontically compromised maxillary incisors with fiber posts and a zirconia all-ceramic system. The materials used for the treatment produced satisfactory esthetic and functional results. PMID- 19548496 TI - American Recovery and Reinvestment Act provisions of interest to dentists. PMID- 19548497 TI - Recruitment drive to start next year. PMID- 19548498 TI - Nurses assess the 48-hour week. PMID- 19548499 TI - Nurses are vital in implementing the osteoporosis care manifesto. PMID- 19548500 TI - Schizophrenia in adults 2: treating acute episodes and promoting recovery. AB - This second in a two-part unit on updated NICE guidance on schizophrenia looks at treating acute episodes and promoting recovery. Part 1 examined detection, assessment and starting treatment. PMID- 19548501 TI - Developing activities to help students achieve learning outcomes in practice placements. AB - The transfer of nurse education into higher education institutions in the late 1990s resulted in the separation of nursing practice and education. Since then, it has often been difficult for student nurses in practice placements to demonstrate that they have achieved their learning outcomes in relation to the assessment tool. Many have asked for support with fulfilling this requirement. Mentors have expressed concern about how best to provide this because ideas on how to gather evidence of what a student is achieving in practice differ significantly. Also, mentors needed help to understand academic jargon used in assessment documentation to ensure equity in the assessment process. This article describes a collaborative project between practice staff and academics to help and support students and mentors when providing evidence that learning outcomes have been achieved in practice placements. PMID- 19548502 TI - Effects of bedrest 1: cardiovascular, respiratory and haematological systems. AB - This is the first in a three-part series on the physiological effects of bedrest. It discusses what happens to the cardiovascular, respiratory and haematological systems when a person is bedridden. Other articles in the series will cover the effects of immobility on the digestive, endocrine, renal, nervous, immune and musculoskeletal systems and will examine the effects of bedrest on the skin. PMID- 19548504 TI - On how staff consultation is vital before service redesign. PMID- 19548503 TI - Helping hard-to-reach families to manage their children's continence problems. AB - AIM: To inform and support the development of new initiatives by the charity ERIC (Education and Resources for Improving Childhood Continence) in the management of paediatric continence problems for disadvantaged families. METHOD: In year 1, in a semi-qualitative pilot study, 45 families of children with continence problems were interviewed. In year 2, we assessed new resources and, in year 3, we reinterviewed 15 respondents from year 1 to measure the impact of the resources. Interviews were conducted with eight new families to provide a comparison. RESULTS: Recruitment to the study was difficult. Parents lacked motivation and self-identification, and had low awareness of the options available. They also reported inappropriate or lack of referral by healthcare professionals. The results also indicated a need for improved understanding of children's continence problems in primary care. CONCLUSION: Where new initiatives were put in place and assessed by parents, all but three children's problems were totally or partially resolved. Resolving problems tended to improve family relationships and emotional stability. PMID- 19548505 TI - Nursing degrees must be built on communication. PMID- 19548506 TI - Continence specialists have the opportunity to inspire services. PMID- 19548507 TI - Nurses must receive sufficient continence education and training. PMID- 19548508 TI - Using bladder ultrasound to detect urinary retention in patients. AB - Bladder ultrasound is now considered a safer alternative to catheterisation in the diagnosis of urinary retention. This article outlines how bladder ultrasound works and its practical uses. PMID- 19548509 TI - A personal account of living with faecal incontinence. PMID- 19548510 TI - Beyond health insurance: public policy to improve health. Introduction. AB - This volume initiates, we hope, a greater focus in the health policy debate on the improvement of health and a departure from the fixation on health insurance. As was observed by Steven Schroeder in the 2007 Shattuck Lecture last year, health care's proportional contribution to premature death is only 10%, compared with behavioral patterns (40%), genetic predisposition (30%), and social circumstances (15%) (Schroeder, 2007). A lesson from the research presented in this volume is that improving health is difficult, far more difficult than simply spending tax dollars to expand health insurance. It is our hope that follow-up work carries this mantle forward in new and innovative ways. PMID- 19548511 TI - Quantifying national spending on wellness and prevention. AB - PURPOSE: We estimate national health expenditures on prevention using precise definitions, a transparent methodology, and a subdivision of the estimates into components to aid researchers in applying their own concepts of prevention activities. METHODOLOGY/APPROACH: We supplemented the National Health Expenditure Accounts (NHEA) with additional data to identify national spending on primary and secondary prevention for each year from 1996 to 2004 across eight spending categories. FINDINGS: We estimate that NHEA expenditures devoted to prevention grew from $83.2 billion in 1996 to $159.8 billion in 2004, in current dollars. As a share of NHEA, this represents an increase from 7.8 percent in 1996 to 8.6 percent in 2004. This share peaked at 9 percent in 2002 and then declined due to reductions in public health spending as a percent of NHEA between 2002 and 2004. Primary prevention represents about half the expenditures, consisting largely of public health expenditures--the largest prevention element. ORIGINALITY/VALUE OF PAPER: Our 2004 estimate that 8.6 percent of NHEA goes to prevention is nearly three times as large as the commonly cited figure of 3 percent, but depends on the definitions used: our estimate falls to 8.1 percent when the research component is excluded, 5.1 percent when consideration is limited to primary prevention plus screening, 4.2 percent for primary prevention alone, and 2.8 percent if we count only public health expenditures. These findings should contribute to a more informed discussion of our nation's allocation of health care resources to prevention. PMID- 19548512 TI - Achieving the Healthy People 2010 goal of elimination of health disparities: what will it take? AB - The second national goal for Healthy People 2010 is the elimination of health disparities related to social disadvantage in the United States. Unfortunately, progress to date has been limited. Our national strategy to achieve this goal has been too narrowly focused on public health. Success will require a broader strategy including alignment of existing national policies in non-health areas that affect the health of the socially disadvantaged such as education, health care, labor, welfare, housing, criminal justice, the environment, and taxation if it is to succeed. Key criteria are needed to begin to prioritize areas for federal investment to achieve this goal. These include the impact of the targeted condition on disparities, evidence base for the intervention, potential impact of the policy on disparities, economic impact, and federal politics. Two "big ideas" offer promise including federal investment in early child education and enhanced primary care within federally qualified community health centers. The proposed criteria are applied to each proposed policy. PMID- 19548513 TI - Influence, information overload, and information technology in health care. AB - We investigate whether information technology (IT) can help physicians more efficiently acquire new knowledge in a clinical environment characterized by information overload. We combine analysis of data from a randomized trial with a theoretical model of the influence that IT has on the acquisition of new medical knowledge. Although the theoretical framework we develop is conventionally microeconomic, the model highlights the non-market and non-pecuniary influence activities that have been emphasized in the sociological literature on technology diffusion. We report three findings. First, empirical evidence and theoretical reasoning suggests that computer-based decision support will speed the diffusion of new medical knowledge when physicians are coping with information overload. Second, spillover effects will likely lead to "underinvestment" in this decision support technology. Third, alternative financing strategies common to new IT, such as the use of marketing dollars to pay for the decision support systems, may lead to undesirable outcomes if physician information overload is sufficiently severe and if there is significant ambiguity in how best to respond to the clinical issues identified by the computer. This is the first paper to analyze empirically and theoretically how computer-based decision support influences the acquisition of new knowledge by physicians. PMID- 19548514 TI - Health disparities and direct-to-consumer advertising of pharmaceutical products. PMID- 19548515 TI - Pharmaceutical innovation and the longevity of Australians: a first look. AB - The purpose of this paper is to examine the impact of pharmaceutical innovation on the longevity of Australians. The approach utilized involves estimation of difference-in-differences models using longitudinal, disease-level data during the period 1995-2003 to determine whether the diseases that had above-average increases in mean vintage (FDA approval year) of drugs had above-average reductions in mortality. Our findings are that the mean age at death increased more for diseases with larger increases in mean drug vintage. A 5-year increase in mean drug vintage is estimated to increase mean age at death by almost 11 months. The number of years of potential life lost before the ages of 65 and 70 (but not before age 75) was reduced by use of newer drugs. During the period 1995 2003, mean age at death increased by about 2.0 years, from 74.4 to 76.4. The estimates imply that, in the absence of any increase in drug vintage, mean age at death would have increased by only 0.7 years. The increase in drug vintage accounts for about 65% of the total increase in mean age at death. Estimated cost per life-year gained from using newer drugs is $10,585. An estimate by previous investigators of the value of a statistical Australian life-year ($70,618) is 6.7 times as large. We acknowledge potential limitations of this study by discussing several reasons why our estimate of the cost per life-year gained from using newer drugs could be too high or low. The value of this paper's evidence is primarily due to the government's Pharmaceutical Benefits Scheme: Australia has much better data on drug utilization than most other countries. PMID- 19548516 TI - Spillover effects of prescription drug withdrawals. AB - Several high-profile prescription drugs have been withdrawn from the U.S. market in the last decade, yet there is no direct evidence of how a prescription drug withdrawal affects consumers' use of remaining drugs within the same therapeutic class. In theory, remaining drugs in the therapeutic class could enjoy competitive benefits or suffer negative spillovers from the withdrawal of a competing drug. Using the Medical Expenditure Panel Survey, we test for spillovers following prescription drug withdrawals in six therapeutic classes between 1997 and 2001. Results vary, but we find stronger evidence of negative spillovers than competitive benefits. We conclude with a discussion of the characteristics of drugs and classes that may influence how remaining drugs are affected by a withdrawal in the class. PMID- 19548517 TI - The psychology of nutrition messages. AB - The purpose of this paper is to explore consumer thinking about nutrition decisions and how firms can use consumers' awareness of the links between nutrients and health generated by public health messages to market products, including ones, which have little nutritional value. We approach this issue by tracking the development of public health messages based on scientific research, dissemination of those messages in the popular press, and use of nutrition claims in food advertisements to assess whether firms are timing the use of nutrition claims to take advantage of heuristic-based decision-making. Our findings suggest that the timing of the development of nutrition information, its dissemination in the press, and use in advertising accords well with a heuristic processing model in which firms take advantage of associations between nutrient information and health in their advertisements. However, the demonstrated relationships may not be causal. Further research will be needed to provide stronger and more comprehensive evidence regarding the proposed message hijacking process. If the message hijacking framework is borne out: (1) simple overall health rating scales could significantly improve consumer decision-making, (2) the impact of misleading advertisements could be mitigated by encouraging a multidimensional view of nutrition, and (3) more intensive regulation of product labeling could limit the impact of hijacked messages. Overall, this paper considers a novel hypothesis about the impact of public health messages on nutrition and health. PMID- 19548518 TI - Evaluation criteria for report cards of healthcare providers. AB - Report cards, performance evaluations, and quality assessments continue to penetrate the lexicon of the healthcare sector. The value of report cards is typically couched as enhancing consumerism among patients, increasing accountability among healthcare providers, and more broadly increasing the transparency of healthcare information. This paper discusses the potential benefits and pitfalls of these performance assessments. This paper briefly reviews empirical evidence regarding the impact of report cards for healthcare providers and synthesizes the role and limitations of these performance measures into distinct evaluation criteria. The rapid proliferation of report cards for healthcare providers suggests a growing need to develop mechanisms and tools to evaluate their impact. The risks associated with utilizing report cards for provider oversight include the deleterious impact on vulnerable populations and a failure to accurately measure quality of care. The capacity to create report cards should not be the sole criterion to develop and utilize report cards to evaluate healthcare providers. Rather, careful consideration of the benefits and risks should accompany the implementation and utilization of report cards into regulatory processes. This report proposes an evaluation checklist by which to assess the role of report cards in a given healthcare context. PMID- 19548519 TI - Evaluating the value of genomic diagnostics: implications for clinical practice and public policy. AB - An important current trend in health care is the move toward personalized medicine. Personalized medicine includes diagnostic and therapeutic interventions, with risk defined through genetics. The key paradigm shift brought about by the advent of personalized medicine is the increased use of in vitro genomic diagnostics. These tests offer the potential of being able to predict which patients are likely to respond to a particular drug, or which patients are likely to develop adverse reactions to a drug. The focus of this paper is the use of genomic diagnostics, and how the increasing development and translation into clinical practice of diagnostic--drug combination products will be adopted into health care delivery. The meaning of value and how to measure it is considered from different perspectives. A novel framework for evaluating the value of genomic diagnostics is proposed. Finally, the implications for regulatory approval and policy are discussed using an illustrative case study. PMID- 19548520 TI - An analysis of risk-taking behavior among adolescent blunt trauma patients. AB - INTRODUCTION: The impact of risk-taking behavior among adolescent blunt trauma patients is not fully appreciated. This study examined the relationship between adolescent risk-taking behaviors, the resultant injury severity, and outcome for blunt trauma. METHODS: Between January 2000 and December 2005, data were collected on adolescent blunt trauma patients (12-18 years) admitted to either a Level I adult trauma center or large urban pediatric hospital. Five groups of risk-taking behavior were examined: ATV riders, drug and alcohol users, unhelmeted motorcyclists, unhelmeted extreme sports participants and unrestrained motor vehicle occupants. Demographic data, mechanism of injury, injury severity, hospital course and outcomes were evaluated for each group. RESULTS: A total of 2030 adolescents were admitted following blunt trauma; 723 adolescents (36%) were engaged in risk-taking behavior at the time of their injury. Most patients were male (68%). Unrestrained MVA occupants were the most frequently encountered risk takers (37%); among this subset, most were unrestrained passengers (74%). Head injuries were frequent (22%) among risk takers. When compared to non-risk-takers, there were no significant age, race, gender, or ISS differences. However, a significantly higher number of positive head CT scans were found among risk taking adolescents (22%, p < 0.05). Mortality was low (3%). CONCLUSIONS: Risk taking behavior is prevalent among adolescent blunt trauma patients. Improved injury prevention strategies are needed to discourage these behaviors during adolescence. PMID- 19548521 TI - Fetal alcohol spectrum disorders (FASD): what medical professionals need to know. PMID- 19548522 TI - Of backpacking, grandchildren, reproduction, and longevity. PMID- 19548523 TI - Protect the skin you are in. PMID- 19548524 TI - [Search for the genes critical for propagation of the prion-like antisuppressor determinant [ISP+] in yeast using insertion library]. AB - The prion-like determinant [ISP+] manifests itself as antisuppressor of certain sup35 mutations. To establish that [ISP+] actually represents a new yeast prion, it is necessary to identify the gene encoding protein corresponding in its prion form to [ISP+]. Analysis of transformants obtained by transformation of [ISP+] strain with insertion gene library revealed three genes controlling the [ISP+] maintenance. These genes are UPF1, UPF2 and SFP1. The SFP1 encodes potenlially prionogenic protein, as it is enriched with asparagine and glutamine residues. Therefore it is the most likely candidate to the role of [ISP+] structural gene. The UPF1 and UPF2 products are components of nonsense-mediated mRNA decay. It was shown that [ISP+] elimination caused by UPF1 and UPF2 inactivation is reversible. It was shown also that Upf1 and Upf2 proteins are not related functionally to Ppzlp phosphatase, influencing [ISP+] manifestation. Possible mechanisms of UPF1 and UPF2 influence on [ISP+] maintenance are discussed. PMID- 19548525 TI - [Molecular variability of testes-specific betaNACtes genes in Drosophila melanogaster genome]. AB - Family of betaNACtes genes in the Drosophila melanogaster genome is a model to investigate the mechanisms of molecular evolution of recently evolved genes. The betaNACtes genes encode proteins, homologous to beta subunit of nascent polypeptide-associated complex (NAC), are expressed in testes and localized on the X chromosome as two two-gene clusters and one separate copy. We collected population polymorphism data for betaNACtes genes using several wild-type stocks of D. melanogaster and compared betaNACtes paralogs with each other. We have shown heterogeneous pattern of betaNACtes genes polymorphism: genes in 3' region of two-gene clusters are low polymorphic, whereas separate betaNACtesl gene is most variable. 5'betaNACtes copies in two-gene tandems are practically identical, whereas 3'betaNACtes copies are highly diverged. Thus, we propose local gene conversion providing selective homogenization of 5'genes. Comparison of betaNACtes paralogs has shown that majority of amino acid differences are in N terminal part of proteins, containing betaNAC domain. McDonald-Kreitman test of betaNACtes paralog divergence shows the involvement of positive selection in the course of betaNACtes gene family evolution. PMID- 19548526 TI - [CENTB5 gene expression in human and mouse]. AB - Centaurin beta5 with unclear function belongs to protein family of centaurins. Human centaurin beta5 is encoded by gene CENTB5 whose intron 14-15 contains low variable minisatellite UPS29, and mouse homolog CENTB5 in analogous intron contains imperfect microsatellite repeat (CATG)19. Earlier we found the association between an occurrence of short UPS29 alleles with some forms of Parkinson disease and epilepsy. Besides this, both human and mice CENTB5 are localized in the same synteny group with SCNN1D and ACOT7 genes which are known to be expressed predominantly in nervous system. Mutations in these genes are connected with neurodegenerative processes and epilepsy. It is known that intra intronic sequences can modulate genes of their location and neighbor and even remote genes. Using RT-PCR we carried out simultaneous analysis of CENTB5, SCNN1D and ACOT7 genes expression. Potential possibility of human intra-intronic tandem repeat UPS29 and of mouse intra-intronic tandem repeat (CATG)19 to regulate/modulate CENTB5, SCNN1D and ACOT7 activity was evaluated in silico. It was found that all these genes were expressed in all studied organs and tissues. It is suggested that minisatellite locus UPS29 can regulate an activity of CENTB5, SCNN1D and ACOT7 in nervous system cells. PMID- 19548527 TI - [Analysis of K-ras, BRCA1/2, CHEK2 mutations and microsatellite markers (loss of heterozygosity at 9p, 17p and 18q) in sporadic pancreas adenocarcinomas]. AB - The purpose of this study was to investigate informativety and clinical significance of most frequent somatic alterations in K-ras, TP53, CDKN2A, MADH4 and more uncommon mutations in BRCA1, BRCA2, CHEK2 genes, which arise on preinvasive stage in sporadic pancreatic adenocarcinomas (PA), in Russian patients. We examined surgically resected and manually microdissected primary PA tissue samples and samples of normal pancreatic tissue for 37 individuals. K-ras mutations in codon 12 were found in 24 tumors (0.65) and none of normal tissue samples. No mutations were detected in BRCA1(185delAG, 300T > G, 4153delA, 4158A > G,5382insC), BRCA2 (695insT, 6174delT) and CHEK2 (1100delC) genes. Informativety for allelic loss of three tumor suppressor genes studied had not statistically significant differences: 60% - for TP53 (GDB186817) and CDKN2A (D9S974 + D9S162); and 65.7% - for MADH4 (D18S363 + D18S474) (t = 0.48). Maximal frequency of loss of heterozygosity (LOH) was observed for CDKN2A - 0.95. For TP53 and MADH4 it was 0.62 and 0.70 respectively. The tumors included 80% cases showing LOH on different chromosomal loci. The combination of K-ras mutations (c.12) and LOH at 9p, 17p and 18q resulted in a high informativety of selected molecular markers: 85.7%. Instability of microsatellites was found only in 9% of PA. PMID- 19548528 TI - [A kinetic study of gamma interferon production in herpes simplex virus-1 DNA prime-protein boost regimen comparing to DNA or subunit vaccination]. AB - The vast majority of the world's population is infected with Herpes simplex virus (HSV). Although antiviral therapy can reduce the incidence of reactivation and asymptomatic viral shedding, and limit morbidity and mortality from active disease, it cannot cure infection. Therefore, the development of an effective vaccine is an important global health priority. In this study, the induction of IFN-gamma production was compared by different herpes simplex virus 1 (HSV1) vaccines. Glycoprotein D (gD1) as a major immunogenic HSV1 glycoprotein was chosen to our study. Balb/c mice were administered with DNA vaccine encoding gD1, subunit glycoprotein vaccine including insect cells infected by a gD1 recombinant Baculovirus, prime DNA vaccine boosted by subunit glycoprotein vaccine, inactivated KOS strain as a positive control, PcDNA3 plasmid and Sf9 cells as a negative control. Evaluation tests showed kinetics of IFN-gamma mRNA at 8, 16 and 32 hours after restimulation sharply decreased whereas, IFN-gamma protein is significantly increased. Our results revealed that at 14 days after immunization IFN-gamma secretion of stimulated cells in all of the vaccinate groups dramatically raised rather than secreted IFN-gamma levels in mice that were analyzed at 7 days after vaccination. In comparison to other groups; Prime-Boost immunization dramatically caused vigorous and prompt IFN-gamma production at 7 days after immunization and 8 hours after restimulation. PMID- 19548529 TI - [Methylation of promoter region of RASSF1A gene and frequencies of allelic imbalances in chromosome 3 critical regions are correlated with progression of clear cell renal cell carcinoma]. AB - The short arm of chromosome 3 (3p) contains several critical regions harboring the set of genes with tumor suppressor activities. The RASSF1A gene (LUCA region, 3p21.31) shows various functions which can be associated with tumorigenesis. Among 3p genes this gene can be most frequently methylated in epithelial tumors of various locations. Here two independent methods (methyl-specific PCR and methyl-sensitive restriction analysis) were used to show significant correlation of methylation level of promoter region of this gene with grade and clinical stage of clear cell renal cell carcinoma (RCC) for the first time. Analysis of 23 polymorphic markers of 3p using the representative set of samples (80 cases RCC), described clinically and histological, permitted to reveal significant correlation between frequency of allelic alterations in some critical regions of 3p (LUCA and AP20) and RCC progression, as distinct from the whole 3p. These data suggest that methylation of promoter region of the RASSF1A gene is associated with RCC progression, and besides, structure-functional alterations in other 3p genes can be also related with RCC progression. In addition, significant correlation between RASSF1A methylation events and allelic losses in the close polymorphic marker was shown here, pointing to the role of "two hit" model for this tumor-suppressor gene inactivation in RCC. PMID- 19548530 TI - [Alteration of SEMA3B gene expression levels in epithelial tumors]. AB - Gene expression decreasing in tumors permits to suggest tumor-suppressor activities for these genes. Thus, mRNA quantity decrease was found for SEMA3B gene in many cell lines of small cell (SCLC) and non-small cell lung carcinoma (NSCLC) and it is well-known that SEMA3B suppresses growth of the NCI-H1299 non small cell lung carcinoma (NSCLC) cell line and tumor formation in nude mice. The aim of this work was to study spectrum of SEMA3B expression level in epithelial tumors of various locations. Using semi-quantitative RT-PCR it was shown for the first time decrease of SEMA3B mRNA quantity (10-250 times as much) in cell lines of renal, breast and ovarian tumors (4/11, 36%). SEMA3B expression profiles in primary tumors of five locations (kidney, lung, breast, ovary and colon) were studied for the first time. This analysis revealed decrease of mRNA quantity (5 1000 times as much) in clear cell renal cell carcinomas with significant high frequency: 25/51, 49% (cases with decrease of mRNA quantity) and 5/51, 10% (cases with increase), P < 0.0001 by Fisher exact test. In addition, the first data about comparatively frequent decrease of mRNA quantity in ovarian (5/16, 31% vs. 2/16, 12%) and colorectal carcinomas (6/11, 54% vs. 2/11,18%) were shown. These results permitted to suggest a possible role of SEMA3B in inhibiting of growth of renal, ovarian and colorectal cancer cells. PMID- 19548531 TI - [Synthesis in Escherichia coli cells and characterization of the active exoribonuclease of severe acute respiratory syndrome coronavirus]. AB - The nsp14 protein, an exoribonuclease of the DEDD superfamily encoded by severe acute respiratory syndrome coronavirus (SARS-CoV), was expressed in fusion with different affinity tags. The recombinant nspl4 proteins with either GST fusion or 6-histidine tag were shown to possess ribonuclease activity but nspl4 with a short MGHHHHHHGS tag sequence at the N-terminus increased the solubility of nspl4 protein and facilitated the protein purification. Mutations of the conserved residues of nspl4 resulted in significant attenuation but not abolishment of the ribonuclease activity. Combination of fluorescence and circular dichroism spectroscopy analyses showed that the conformational stability of nsp14 protein varied with many external factors such as pH, temperature and presence of denaturing chemicals. These results provide new information on the structural features and would be helpful for further characterization of this functionally important protein. PMID- 19548532 TI - [Design of oligonucleotide inhibitors of the human DNA-methyltransferase 1]. AB - Mammalian DNA methyltransferase 1 (Dnmt1) is responsible for copying DNA methylation patterns during cell division. A number of studies demonstrate that Dnmt1 plays an important role in carcinogenesis, that causes, in particular, significant interest in searching for specific inhibitors of this enzyme. In the present study, with the purpose of design of oligonucleotide inhibitors of human Dnmt1, a number of single-, double-stranded and hairpin DNA-structures, containing canonical or modified enzyme recognition site 5'-CG were constructed on the basis of uniform 22 b sequence. It was shown, that such structural features as C:A-mismatch, phosphorothioates and hairpin are capable to incrementally increase oligonucleotide affinity to Dnmt1. The improvement of inhibitor properties were also achieved by substitution of target cytosine with 5,6-dihydro-5-azacytosine, 5-methyl-2-pyrimidinone and 6-methyl-pyrrolo-[2,3-d]-2 pyrimidinone. The concentrations of the most efficient oligonucleotides caused 50% inhibition of methylation of 1 microM conventional DNA substrate, polymer poly(dI-dC) * poly(dI-dC), were about 10(-7) M. In the equal in vitro conditions the constructed oligonucleotide inhibitors demonstrated much stronger effect compared to known inhibitors of Dnmt1, which were used as controls. PMID- 19548533 TI - [Search for an optimal orientational ordering of Qy transition dipoles of subantennae molecules in superantenna of photosynthetic green bacteria. Model calculations]. AB - This work continues a series of our investigations on efficient strategies of functioning of natural light-harvesting antennae, initiated by our concept of rigorous optimization of photosynthetic apparatus structure by functional criterion. Using computer modeling for the functioning of the natural antennae, we suggested some basic principles for designing optimal model systems. Targeted searches for these principles in in vivo systems allowed us to recognize some of them in natural antennae. This work deals with the problem of the structure optimization of nonuniform superantennae of photosynthetic green bacteria. These superantennae consist of several uniform subantennae which produces a problem of their optimal coordination. In this work, we used mathematical modeling for the functioning of these natural superantennae to consider a possible way to optimize the superantenna structure using optimization of mutual spatial orientation of Qy transition dipoles of subantennae pigments. This allowed us to determine some modes of optimal orientational ordering of Qy transition dipoles of subantennae pigments in the model of the green bacterium Chloroflexus aurantiacus superantenna. It was shown that the optimal mutual orientation of Qy transition dipoles of subantennae pigments (resulting in stable minimizing of the energy transfer time within the superantenna and, as a consequence, in decrease in energy losses) ensures the high efficiency and stability of the superatenna functioning. PMID- 19548534 TI - [Design and structural and thermodynamic studies of a chimeric protein derived from spectrin SH3-domain]. AB - A number of chimerical constructions based on the spectrin SH3 domain were designed for structural and thermodynamic studies of protein folding and protein ligand interactions. SH3 domains were found in many regulatory proteins and operate through weak interactions with proline-rich fragments of the partners. The recombinant protein studied in this work (WT-CIIA) was constructed by linking the peptide PPPVPPYSAG to the domain C-terminal trough a long 12-residue linker with the aim to achieve stable ligand binding in orientation II, which until now has not been considered as typical for spectrin domain. A comparison of fluorescence spectra of the chimerical protein and the parent domain suggests that the ligand sticks to the conservative binding site. The analysis of the urea induced unfolding curves revealed, however, that the protein-ligand contact is not stable enough and as a result the chimerical protein structure unfolds in two steps. In order to clarify the structural aspects of the protein-ligand interaction, WT-CIIA was crystallized and a set of the X-ray diffraction data at 1.75 angstroms resolution was acquired. Preliminary analysis of the diffraction data indicated that the crystals belong to the space group P32, with unit-cell parameters a = b = 3639, c = = 112.17 angstroms, alpha = beta = 90.0, gamma = 120.0. PMID- 19548535 TI - [Liquid-crystalline dispersions formed by complexes of linear double-stranded DNA molecules with dendrimers]. AB - The formation of liquid-crystalline dispersions as a result of interaction of linear, double-stranded DNA molecules with poly(propyleneimine) dendrimers in water-salt solutions was studied. It was shown, that this process does not depend on the ionic strength of solution and molecular structure of dendrimer. By means of the atomic force microscopy, it was established, that in the case of the dendrimer molecules of the 4th generation (G4), the mean size of particles of (DNA-dendrimer G4) liquid-crystalline dispersion is equal to 300-400 nm. The "boundary" conditions (ionic strength of solutionand molecular mass of dendrimer) of formation of optically active (cholesteric) and optically inactive of the (DNA dendrimer) dispersions were determined using circular dichroism spectroscopy. The interaction of dendrimers of 1st, 2nd, 3rd and 5th generations with DNA molecules results in the obtaining of the optically inactive dispersions. Dendrimer molecules of 4th generation induce the formation of two types dispersions: in solutions of high ionic strength (micro > 0.4) they induce the formation of cholesteric liquid-crystalline dispersions, and in solutions of low or intermediate ionic strength (micro < 0.4) they can form the optically inactive one. The "molecular crowding" affects both the efficiency of binding of dendrimer molecules of 4th generation to DNA, and the mode of spatial packing of (DNA dendrimer G4) complexes in particles of liquid-crystalline dispersion. The possible reasons capable of explaining the structural polymorphism of (DNA dendrimer) liquid-crystalline dispersions are discussed. PMID- 19548536 TI - [Cloning, expression and comparative analysis of peroxiredoxine 6 from different species]. AB - Human, rat, Xenopus and Drosophila (Dpx2540 and Dpx6005) cDNA of peroxiredoxins were cloned and expressed in Escherichia coli. Their enzymatic activity, temperature optimum and thermostability were determined. For H2O2 the activity of enzymes decreased in the following order: DPx2540 > > human > Xenopus >rat > DPx6005. For tret-butyl hydroperoxide the order of activity decrease is: DPx2540 = DPx6005 > rat > Xenopus > human. Effectiveness of plasmid DNA protection from oxidative damage mediated by Fenton reaction is: Dpx2540 > Dpx6005 = rat = human > Xenopus. The optimal temperature for activity of all these enzymes is 37 degrees C. Peroxiredoxins from rat, Xenopus and Drosophila (Dpx 6005) retain no less than 50% of activity in a wide temperature range (10-50 degrees C) in contrast to human and Drosophila (Dpx 2540) enzymes with the interval of only 25 45 degrees C. The thermostability of enzymes decreased in the following order: Dpx6005 > or = rat > human > Xenopus > Dpx2540. So, there is negative correlation between activity and stability of peroxiredoxin 6. PMID- 19548538 TI - [Novel structural tree for (alpha + beta)-proteins containing abCd-units]. AB - A database of 926 (alpha + beta)-proteins and (alpha + beta)-domains containing abCd-units (among them 401 are nonhomologous) has been compiled from the Protein Data Bank (total 2636 PDB entries). A novel structural tree for this structural class of proteins that is composed of 286 possible polypeptide chain folds has been constructed. The structural classification of (alpha + beta)-proteins containing abCd-unit based on the structural tree has been developed. Both the database and the structural tree are accessible at the web-site (http://strees.protres.ru/). PMID- 19548537 TI - [Prognosis of affinity change of the TATA-binding protein to TATA-boxes upon polymorphisms of the human gene promoter TATA boxes]. AB - TATA-binding protein (TBP) is a subunit of basal transcription factor TFIID that recognizes and binds to the TATA-box on TATA-containing promoters of class II genes, and starts assembling RNA polymerase II basal transcription complex. It is shown in many works that the sequence of TATA-box with its flanking regions affects the level of basal and activated transcription. TATA-box polymorphisms and human hereditary diseases associated with them show that TBP/TATA interaction may indirectly affect gene regulation in vivo. The object of this work is to determine changes in the TBP/TATA affinity upon polymorphisms in TATA-boxes of human gene promoters. We assess changes in TBP/TATA affinities in silico by using our formula of equilibrium TBP/TATA binding upon four consecutive steps: nonspecific binding <--> sliding <--> braking (stopping) <--> stabilization. Our prognoses agree with known examples of TATA-box polymorphisms and human hereditary diseases associated with them. PMID- 19548539 TI - [Modeling evolution of regulatory signals for gene expression in bacteria]. AB - A model of evolution of a regulatory signal along the phylogenetic tree of species taking into account the secondary RNA structure is suggested. Based on this model, an algorithm is presented. It inputs the extant primary structure of a signal for the leaves of the phylogenetic tree and computes the primary and secondary structures of all the nodes. Another result of the algorithm is a multiple alignment of extant sites of a regulatory signal taking into account the secondary structure of the signal. The algorithm ha s been implemented and successfully tested on biological data representing three types of regulation in bacteria. PMID- 19548540 TI - [Establishment of a novel sensitive method for detecting methylation modification on DNA of Escherichia cell]. AB - This study focused on finding a novel sensitive method to determine the methylation modification at DNA dam (GATC) sites in Escherichia coli. A new plasmid which contained three GATC sites recognized by restriction enzyme BclI and one GAATTC site recognized by EcoRI was transformed into E. coli stains AB1157 (dam+) and GM2929 (dam-) respectively. Then the plasmid DNA was digested by restriction enzyme Bdl (T*GATCA), which was sensitive to methylation. The results showed that the plasmid derived from AB1157 was not digested while that from GM2929 was, for the methylation level of the former was high while the latter was low. So by detecting the methylation of plasmid transferred into the strain, we could determine whether methylation existed at DNA dam (GATC) site in E. coli. This method was effective and rapid; moreover, the digested fragments were not dispersive. It also made a basis for the detection of whether methylation occurred in mode beings by low-energy ion beam. PMID- 19548541 TI - [New method of construction of artificial translational-coupled operons in bacterial chromosome]. AB - The new method of translational-coupled operons construction in bacterial chromosome has been developed on the basis of recombineering approach. It includes construction in vitro of the artificial operon with efficiently translated proximal cistron followed by its insertion E. coli chromosome, modification of the operon due to Red-driven insertion of the special "Junction" with excisable selective marker in the intercistronic region of the initial operon and excising the marker. The structure of this Junction has been designed and tested in the present investigation. It consists of: 1) E. coli rplC-rplD intercistronic region for placing the TAA-codon of the proximal operon's gene in the SD-sequence (TAAGGAG) of rplD; 2) Cm(R)-gene flanked by lambdaattL/R-sites in such a fashion that after lambdaInt/Xis-driven excision of the marker the residual lambdaattB-site would not contain the termination codons in frame with ATG of rplD; 3) E. coli trpE-trpD intercistronic region for location of ATG of trpD at the position of initiation codon of the distal gene of original operon. The general design of desired construction provides the conversion of the original two-cistronic operon into three-cistronic operon with translational coupled genes, where the coupling of the artificial ORF (rplD'-lambdaattB-'trpE) with the proximal gene is occurred due to rplC-rplD intercistronic region and the coupling of this ORF with the distal gene--due to trpE-trpD. The experimental implementation of the described strategy was showed by construction of artificial operon P(tac-aroG4-serA5, where expression optimization of the distal serA5 gene was achieved via construction of three-cistronic operon with translational coupled genes. PMID- 19548542 TI - [Efficient approach for potato viral pathogen sensitive diagnostic and identification]. AB - Potato, one of the most widespread agricultural plants in Russia, is strongly affected by various pathogens of viral, bacterial, and fungal origin as well as by pests. Their simple and accurate diagnostics and identification sound rather important both for production of virus free planting material and to perform monitoring of the phytosanitary state of planting areas. Based on qualitative Fluorescent Amplification--based Specific Hybridization Polymerase Chain Reaction (FLASH-PCR) we have developed the diagnostic systems, which provided fast, careful, and with the minimum risk of contamination in the working zone by amplification products, detection of the major potato pathogens, i. e. A, Y, X, M, S potato viruses, potato leafroll virus, potato mop top virus, as well as potato spindle tuber viroids. PMID- 19548543 TI - [Increased efficiency of recombinant proteins production in plants due to optimized translation of RNA of viral vector]. AB - One of the most efficient methods for fast and efficient production of the target proteins in plants is based on the use of self-replicating recombinant viral vectors. We constructed phytoviral vector based on the genome of potato X virus containing the sequence of 5'-untranslated region of RNA 4 of alfalfa mosaic virus immediately upstream of the target gene. We demonstrated that incorporation of this sequence into the viral vector results in 3-4 fold elevation of the level of production of the target protein in plant due to increased efficiency of translation of viral subgenomic RNA comprising the target gene. The new vector may be used for production of recombinant proteins in plants. PMID- 19548544 TI - [Alternative splicing tends to involve phosphorylation sites]. PMID- 19548545 TI - Introduction to the economics of obesity. PMID- 19548546 TI - Endogenous food quality and bodyweight trend. AB - This paper follows previous economic studies on bodyweight by looking at bodyweight as individuals' choices in response to changes in income and food prices. However, it goes one step further to add another relevant choice under individual control: food quality. It shows that the upward trend of bodyweight caused by economic growth and technological innovations may well be self-limiting in the sense that the bodyweight growth in the future is likely to be slower or reversed In particular, it finds that much of further income growth will be used for improving food quality rather than increasing caloric intake. Moreover, further technological innovations that focus on lowering the price of high quality (healthy) food relative to that of low-quality food would encourage substituting food quality for food quantity (calories). PMID- 19548547 TI - Access to fast food and food prices: relationship with fruit and vegetable consumption and overweight among adolescents. AB - We examine the extent to which food prices and restaurant outlet density are associated with adolescent fruit and vegetable consumption, body mass index (BMI), and the probability of overweight. We use repeated cross-sections of individual-level data on adolescents from the Monitoring the Future Surveys from 1997 to 2003 combined with fast food and fruit and vegetable prices obtained from the American Chamber of Commerce Researchers Association and fast food and full service restaurant outlet density measures obtained from Dun & Bradstreet. The results suggest that the price of a fast food meal is an important determinant of adolescents' body weight and eating habits: a 10% increase in the price of a fast food meal leads to a 3.0% increase in the probability of frequent fruit and vegetable consumption, a 0.4% decrease in BMI, and a 5.9% decrease in probability of overweight. The price of fruits and vegetables and restaurant outlet density are less important determinants, although these variables typically have the expected sign and are often statistically associated with our outcome measures. Despite these findings, changes in all observed economic and socio-demographic characteristics together only explain roughly one-quarter of the change in mean BMI and one-fifth of the change in overweight over the 1997-2003 sampling period. PMID- 19548548 TI - A behavioral model of cyclical dieting. AB - This paper presents a behavioral economics model with bounded rationality to describe an individual's food consumption choices that lead to weight gain and dieting. Using a physiological relationship determining calories needed to maintain weight, we simulate the food consumption choices of a representative female over a 30-year period. Results show an individual will periodically choose to diet, but that diet will reduce weight only temporarily. Recurrence of weight gain leads to cyclical dieting, which reduces the trend rate of weight increase. Dieting frequency is shown to depend on decision period length, dieting costs, and habit persistence. PMID- 19548549 TI - Effects of Title IX and sports participation on girls' physical activity and weight. AB - In this study, we examined the association between girls' participation in high school sports and the physical activity, weight, body mass, and body composition of adolescent females during the 1970s when girls' sports participation was dramatically increasing as a result of Title IX. We found that increases in girls' participation in high school sports, a proxy for expanded athletic opportunities for adolescent females, were associated with an increase in physical activity and an improvement in weight and body mass among girls. In contrast, adolescent boys experienced a decline in physical activity and an increase in weight and body mass during the period when girls' athletic opportunities were expanding. Taken together, these results strongly suggest that Title IX and the increase in athletic opportunities among adolescent females it engendered had a beneficial effect on the health of adolescent girls. PMID- 19548550 TI - Obesity and diabetes: the roles that prices and policies play. AB - The entrance of economics into the literature on obesity and diabetes has been instrumental in showing how people respond to incentives when maximizing their health. In this paper some of the roles that prices and policies have played in the surging obesity and diabetes rates across the world are addressed. The paper focuses on the possible role that prices of foods with high glycemic indexes play in determining blood sugar levels, and addresses the recent concern with high fructose corn syrup and genetically modified goods across the world. The possible links and implications suggest that future research in the area is urgently needed. PMID- 19548551 TI - Advances in bariatric surgery for obesity: laparoscopic surgery. AB - CONTEXT: The most advanced and fastest growing form of bariatric surgery is laparoscopic gastric bypass. Very little is known about population-based 180-day laparoscopic bypass costs, complication rates, readmission rates, and post operative care. OBJECTIVE: To examine the 6-month costs and outcomes of laparoscopic vs. open bariatric bypass surgery using a national population-based sample. DESIGN: We use the 1998-2003 Nationwide Inpatient Sample to examine national trends in the rate of laparoscopic bypass. To examine postoperative outcomes, we examine insurance claims for 2,384 bariatric bypass surgeries, at 308 hospitals, among a population of 5.6 million non-elderly people covered by large employers across 49 states in 2001 and 2002. Multivariate logit regression analysis is performed to risk-adjust outcomes. MAIN OUTCOME MEASURES: 180-day outcomes: 12 complications specific to bariatric surgery and 44 general post operative conditions, readmission rates, ER rates, and expenditures following bariatric surgery. RESULTS: Between 1998 and 2003, the national percentage of bariatric bypass surgeries that were laparoscopic grew from 1.5 to 17.1%. There was no significant difference in in-hospital mortality between laparoscopy and open surgery. With the 2001-2002 claims data, we find that of the patients having bypass surgery, men had 48% lower odds of having laparoscopy and that high bariatric volume hospitals were close to four times more likely to use laparoscopy. Laparoscopic bypass, compared with open bypass, had 34% lower odds of a complication during the initial surgical stay, 27% lower odds of a 30-day complication, but no statistically significant difference in 180-day complications. Laparoscopy had 49% higher odds of having the general 44 post operative conditions, with 45% higher odds of a readmission and 54% higher odds of an ER visit. However, overall, laparoscopy resulted in a 23% lower number of hospital days and 9% lower 180-day expenditures. CONCLUSION: The laparoscopic cost-savings during the less invasive initial surgery stay outweigh the increase in post-discharge utilization. Further cost-savings will only emerge from laparoscopy only if its late post-operative complications are reduced. More cost savings will also emerge as more physicians switch to the use of laparoscopy for bypass surgery. PMID- 19548552 TI - One pill makes you smaller: the demand for anti-obesity drugs. AB - The doubling of obesity in the U.S. over the last 25 years has led policymakers and physicians to encourage weight loss, but few methods of weight loss are effective. One promising avenue is pharmacotherapy. However, little is known about the use of anti-obesity drugs. This paper describes the market for anti obesity drugs and studies the utilization of anti-obesity drugs using data from the Medical Expenditure Panel Survey for 1996-2002, a period that is interesting because it covers the introduction of three, and the withdrawal of two, anti obesity drugs from the market. Our results point to wide sociodemographic disparities in anti-obesity drug use. Women are almost 200% more likely than men to use anti-obesity drugs. Hispanics and African-Americans are only 39% as likely as Whites to use them. Those with prescription drug coverage are 46% more likely to use anti-obesity drugs. We also find that the vast majority of subjects who are approved to take these drugs are not taking them, and a significant number who are not approved to take the drugs are taking them. We find strong evidence that the well-publicized 1997 withdrawal of fenfluramine and dexfenfluramine had a chilling effect on the overall market for anti-obesity drugs. We find little difference in observed characteristics between those who took the withdrawn drugs and those who took the other anti-obesity drugs in the market. PMID- 19548553 TI - Obesity, employment and wages in Europe. AB - This paper examines the associations between obesity, employment status and wages for several European countries. Our results provide weak evidence that obese workers are more likely to be unemployed or tend to be more segregated in self employment jobs than their non-obese counterparts. We also find difficult to detect statistically significant relationships between obesity and wages. As previously reported in the literature, the associations between obesity, unemployment and wages seem to be different for men and women. Moreover, heterogeneity is also found across countries. Such heterogeneity can be somewhat explained by some labor market institutions, such as collective bargaining coverage and employer-provided health insurance. PMID- 19548554 TI - Obesity and occupational attainment among the 50+ of Europe. AB - This paper brings a European perspective to the mainly U.S.-based literature on the relationship between obesity and labour-market outcomes. Using micro-data on workers aged 50 and over from the newly developed SHARE database, the effects of obesity on employment, hours worked, and wages across 10 European countries were analysed. Pooling all countries, the results showed that being obese was associated with a significantly lower probability of being employed for both women and men. Moreover, the results showed that obese European women earned 10% less than their non-obese counterparts. For men, however, the effect was smaller in size and insignificant. Taking health status into account, obese women still earned 9% less. No significant effect of obesity on hours worked was obtained, however. Regressions by country-group revealed that the effects of obesity differed across Europe. For instance, the effect of obesity on employment was greatest for men in southern and central Europe, while women in central Europe faced the greatest wage penalty. The results in this study suggest that the ongoing rise in the prevalence of obesity in Europe may have a non-negligible effect on the European labour market. PMID- 19548555 TI - Gender, body mass, and socioeconomic status: new evidence from the PSID. AB - Previous research provides evidence of a negative effect of body mass on women's economic outcomes. We extend this research by using a much older sample of individuals from the Panel Study of Income Dynamics and by using a body mass measure that is lagged by 15 years instead of the traditional 7 years. One of the main contributions of this paper is a replication of previous research findings given our differing samples and measures. We compare OLS estimates with sibling fixed effects estimates and find that obesity is associated with an 18% reduction in women's wages, a 25% reduction in women's family income, and a 16% reduction in women's probability of marriage. These effects are robust--they persist much longer than previously understood and they persist across the life course, affecting older women as well as younger women. PMID- 19548556 TI - Health insurance and the obesity externality. AB - If rational individuals pay the full costs of their decisions about food intake and exercise, economists, policy makers, and public health officials should treat the obesity epidemic as a matter of indifference. In this paper, we show that, as long as insurance premiums are not risk rated for obesity, health insurance coverage systematically shields those covered from the full costs of physical inactivity and overeating. Since the obese consume significantly more medical resources than the non-obese, but pay the same health insurance premiums, they impose a negative externality on normal weight individuals in their insurance pool. To estimate the size of this externality, we develop a model of weight loss and health insurance under two regimes--(1) underwriting on weight is allowed and (2) underwriting on weight is not allowed. We show that under regime (1), there is no obesity externality. Under regime (2), where there is an obesity externality, all plan participants face inefficient incentives to undertake unpleasant dieting and exercise. These reduced incentives lead to inefficient increases in bodyweight, and reduced social welfare. Using data on medical expenditures and bodyweight from the National Health and Interview Survey and the Medical Expenditure Panel Survey, we estimate that, in a health plan with a coinsurance rate of 17.5%, the obesity externality imposes a welfare cost of about $150 per capita. Our results also indicate that the welfare loss can be reduced by technological change that lowers the pecuniary and non-pecuniary costs of losing weight, and also by increasing the coinsurance rate. PMID- 19548557 TI - Obesity, hospital services use and costs. AB - OBJECTIVE: To quantify the association between obesity and somatic hospital costs and number of overall somatic hospital contacts--number of inpatient admissions, number of outpatient visits, and number of emergency department visits--based on anthropometric measurements of waist circumference (WC) and information from The National Patient Registry and The Danish Case-Mix System (DRG). PARTICIPANTS: The study population consisted of two sub-samples from the Inter99 study at Research Centre for Prevention and Health in 1999-2001. One sub-sample used WC as an indicator for obesity (n = 5,151), whereas the other used BMI as an indicator for obesity (n = 4,048). Using WC, obesity was defined as WC > 102 cm for men and > 88 cm for women. Normal weight was defined as circumference < 94 cm for men and < 80 cm for women. Using BMI, obesity was defined as BMI > 30 kg/m2, whereas individuals with BMI = 18.5-24.9 kg/m2 were defined as normal weight. Individuals with BMI < 18.5 kg/m2 were excluded from both sub-samples. DESIGN: We undertook a 3-year retrospective study of the relationship between obesity and use of hospital resources. Data on hospital contacts and costs were obtained from The National Patient Registry and DRG. Analyses were performed using two-part models and Poisson regression. Outcome variables were costs and hospital contacts. RESULTS: This study has demonstrated that obese individuals have a greater use of hospital services and greater hospital costs compared with normal weight individuals. When using WC as an indicator for obesity, mean hospital costs were 33.8% greater among obese women and 45.3% greater among obese men in a 3-year period but the differences were not significant. When using BMI to measure obesity, obese men had significantly greater costs (57.5%) than normal weight men. Furthermore, obese men and women (indicated by WC) had an increased number of hospital contacts compared with normal weight individuals (rate ratio 1.32, 95% CI 1.21-1.43 for men and 1.20, 95% CI 1.11-1.28 for women) including inpatient admissions, outpatient visits, and emergency department visits. The same trends were seen when obesity was indicated by BMI. PMID- 19548558 TI - How much does obesity matter? Results from the 2001 Canadian Community Health Survey. AB - A five category self-reported health indicator together with the self-reported prevalence of diabetes and heart disease for older Canadians, are examined using data from five cohorts of men and women from the 2001 Canadian Community Health Survey. Consistent with other studies we find that smoking and dietary behaviors are highly correlated with general self-reported health, diabetes, and heart disease. Individual standardized weight, the body mass index, was negatively associated with health outcomes for all age groups, but became less important with age as socioeconomic variables became more important. Socioeconomic variables explained more of the variation in health outcomes than the combined effects of tobacco use and excessive weight problems. In addition, there is compelling evidence that obesity could overtake smoking as the leading cause of health problems in Canada. PMID- 19548559 TI - [Expression of PI3K/Akt signal pathway in acute myocardial ischemia]. AB - OBJECTIVE: To explore the influence of the PI3K/Akt-mediated signal pathway in acute myocardial ischemia (AMI) rats, the expression of pAkt, Akt, Caspase3 and p38 in myocardium and somatic muscles of AMI rats were detected. METHODS: The rats model of AMI were established by peritoneal injecting isoprinosine (ISO), and were detected by electrocardiogram and haemodynamics. The expressions of pAkt, Caspase3 and p38 in somatic muscles and cardiac muscles of AMI and normal rats were detected by Dot blot hybridization and Western blot. RESULTS: In contrast with normal rats, electrocardiogram of AMI rats showed a lower displacement of ST segment (> or = 0.1 mv). The expression of pAkt, Caspase3 and p38 were higher than those in normal rats (P < 0.05). No apparent changes were observed in expression of Akt (P = 0.477). CONCLUSION: Expressions of pAkt, Akt and correlated apoptosis molecule Caspase3 and p38 in cardiac and somatic muscles of AMI rats were higher than those in normal rats. No apparent changes were observed in expression of Akt. PMID- 19548560 TI - [Influence of PAHs exposure and GSTT1, GSTM1 genotypes on urinary 1-OHP as exposure biomarker]. AB - OBJECTIVE: To study the influence of polycyclic aromatic hydrocarbons (PAHs) exposure (ambient concentration and smoking) and GSTT1, GSTM1, genotypes on urinary 1-OHP as exposure biomarker. METHODS: 51 Cops who working outside were selected as study group, 48 Cops who working in office were selected as control Group. The ambient concentrations of PAHs both of two circumstances and the urinary 1-OHP of subjects were measured. The ambient concentrations of PAHs and the urinary 1-OHP were determined by high performance liquid chromatography (HPLC). The GSTT1, GSTM1 genotypes were determined by PCR. RESULTS: The ambient concentrations of PAHs in the study group and the control group were 20.85 ng/m3 and 12.79 ng/m3 respectively. There were not significant difference in urinary 1 OHP among the persons who had the deference GSTT1, GSTM1, genotypes and the same smoking habits in the study group and the control group. The urinary 1-OHP concentrations of the study group were higher than those of the control group when they had the same genotypes for non-smokers. Smoking stratified analysis showed that urinary 1-OHP concentrations of smokers were higher than those of non smokers, the urinary 1-OHP concentrations of smokers in control group were higher than those of the non-smokers in study group. CONCLUSION: The ambient PAHs concentration and smoking could be the important factors generated the urinary 1 OHP concentration. While the levels of ambient PAHs were low, the smoking could change more prominent. The GSTT1, GSTM1 genotypes could not be the major influential factors. PMID- 19548561 TI - [Study of serotonin transporter gene polymorphisms in disease of patients with abnormal black Savda]. AB - OBJECTIVE: To explore the association between patients with abnormal black Savda according to Uighur medicine theory and the serotonin transporter (5-HTT) gene polymorphisms in the promoter (5-HTTLPR) and the variable number tandem repeats (VNTRs) in intron 2 (5-HTTVNTR). METHODS: Divided the patients into four body fluids according to Uighur medicine theory, PCR was used to detect genotype and allele frequencies of 5-HTILPR and 5-HTTVNTR polymorphisms in 66 patients with abnormal black Savda (including 35 cases of hypertension, 19 cases of diabetes, 12 cases of cancer diagnosis) and 45 control subjects. RESULTS: No significant differences for the genotype frequency or the allele frequency of 5-HTTLPR and 5 HTTVNTR were revealed between patients with abnormal black Savda and control subjects. More control subjects tended to be of L/L genotype (15.56%) than hypertension with abnormal black Savda (5.72%). On the contrary, more cancer diagnosis with abnormal black Savda tended to be of L/L genotype (25.00%) and L (54.17%) allele than control subjects L/L genotype (15.56%) and L allele (37.78%). On the other hand, fewer control subjects tended to be of 10/10 genotype (4.45%) than diabetes with abnormal black Savda (10.53%) and cancer diagnosis with abnormal black Savda (8.34%). CONCLUSION: 5-HTTLPR L/L genotype may be a protective factor for hypertension with abnormal black Savda, 5-HTTLPR L/L genotype and L allele may be risk factors for cancer diagnosis with abnormal black Savda, 5-HTTVNTR 10/10 genotype may be risk factors for cancer diagnosis with abnormal black Savda and diabetes with abnormal black Savda. PMID- 19548562 TI - [Damage on lung and systemic inflammation induced by nickel sulfate intratracheal instillation in rats]. AB - OBJECTIVE: To investigate the acute toxic effects induced by nickel sulfate (NiSO4) in order to clarify the possible role of nickel in PM2.5. METHODS: Twenty eight male Wistar rats were divided into four groups, NiSO4 was administered by intratracheal instillation at the doses of Ni 0, 7.5, 75 and 750 microg/kg for 24h respectively. The rats were sacrificed and bronchoalveolar lavage fluid (BALF) were collected and analyzed to estimate the injury (total protein (TP) and lactate dehydrogenase (LDH)) and inflammation of lung. Blood were collected and parameters of inflammation (total cell numbers, interleukin-6 (IL-6), tumor necrosis factor (TNF-alpha)) and CRP were measured. The nickel levels in the blood were also determined. RESULTS: NiSO4 induced significant increases of leukocytes, neutrophil, TP contents and LDH activity in BALF (P < 0.05, P < 0.01). The level of TNF-alpha and IL-6 in BALF of NiSO4-treated rats increased significantly compared with those of the control group (P < 0.05). The level of TNF-alpha in blood of NiSO4-treated rats increased significantly compared with the control group. Concentrations of CRP in the serum were more higher in NiSO4 treated rats than the control group, but statistical significances were only seen in the highest dosage group (Ni 750 microg/kg). Nickel in serum of NiSO4-treated rats increased significantly in a dose-dependent manner. CONCLUSION: NiSO4 induced pulmonary and systemic inflammation effects in rats, suggested that NiSO4 may play an important role in the acute effects of PM2.5. PMID- 19548564 TI - [Relationship between genetic polymorphisms of RFC1 A80G and nonsymdromic cleft lip with or without palate]. AB - OBJECTIVE: To explore the relationship between genetic polymorphisms of reduced folate carrier (RFC)1 A80G and nonsymdromic cleft lip with or without palate on Chinese population. METHODS: There were 97 NSCL/P case-parent triads those were selected as case group. At the same period, 104 healthy subjects were selected together with their biological parents as control group. For all subjects the polymorphisms of RFC1 A80G were examined by PCR-RFLP method. RESULTS: There was no statistical difference in genotype and gene frequencies for RFC1 A80G variants among family members between case group and control group in offsprings and fathers and mothers (P < 0.05). The odds ratio (OR), confidence interval (CI) and P value of offspring, father and mother genotype (AG) were 0.87 (0.44-1.70), 0.657; 1.09 (0.54-2.21), 0.788; 1.63 (0.79-3.36), 0.152 respectively. The OR, CI and P value of offspring, father and mother allele (GG) were 0.48 (0.19-1.23), 0.094; 0.93 (0.38-2.23), 0.850; 1.30 (0.46-3.67), 0.584 respectively. The OR, CI and P value of offspring, father and mother allele (G) were 1.22 (0.78-1.94), 0.386; 1.02 (0.64-1.61), 0.945; 0.91 (0.58-1.41), 0.660. The G allele could not increase the risk of NSCL/P. But in the transmission disequilibrium test (TDT) analysis, the transmission of the G allele was 40 times, the A allele was 71 times, the A allele was more likely to transmit to the sicken offspring (chi-2 = 8.658, P < 0.05). Results of haplotype-based haplotype relative risk (HHRR) analysis (chi-2 = 10.31, P < 0.05) and family-based association tests (FBAT) ( Z = 2.942, P < 0.05) were showed that there was an association between RFC1 A80G variant and the risk of NSCL/P. CONCLUSION: The statistical analysis of nuclear family could evidence of linkage in the presence of disequilibrium, there was an association between RFC1 A80G variant and the risk of NSCL/P, and the A allele could have an association with the dominant high-risk of NSCL/P. PMID- 19548563 TI - [Dimethylarsinous acid-promoted skin tumorigenesis through the induction of oxidative stress in mice]. AB - OBJECTIVE: To investigated the relationship between skin-tumor promotion and oxidative stress caused by dimethylated arsenic in mice. METHODS: The experimental animal model was used to examine the effect of dimethylated arsenic, a metabolite of DMA(V), dimethylarsinous acid (DMA(III)) in skin tumorigenesis in mice. The 8-oxo-2'-deoxyguanosine (8-oxodG) analysis of epidermis was based on the method of HPLC. RESULTS: When mice were topically treated with trivalent dimethylated arsenic (DMA(III)), a further reductive metabolite of DMA(V), not only an increase in skin tumors but also an elevation of 8-oxodG in epidermis were observed. CONCLUSION: These results suggest that tumor promotion due to DMA(V) administration is mediated by DMA(III) through the induction of oxidative stress. PMID- 19548565 TI - [Research on the evaluation methods for antioxidant capacity of several bioactive substances in vitro]. AB - OBJECTIVE: To study the evaluation methods for antioxidant capacity of bioactive substance in vitro, and to apply the methods which is established in physicochemical environment in order to give the preliminary assessment of test subjects. METHODS: The combined application of ABTS and FRAP methods in the assessments, using the UV-VIS spectrophotometer and the autoanalyzer respectively test the antioxidant capacity of quercetin, curcumin, DL-alpha-tocopherol and procyanidine at 734 nm in ABTS and at 595 nm in FRAP. RESULTS: ABTS: The TEAC values of quercetin and curcumin were about 2.02 and 0.50. 1 g DL-alpha tocopherol and anthocyanins were equivalent to 2.06 mmol, 2.897 mmol of Trolox in scavenging free radicals capacity. FRAP: Used 1.0 mmol/L FeSO4 as the reference standard, quercetin, curcumin and Trolox equivalent molar about 5.73, 1.18 and 2.09. DL-alpha-tocopherol, antioxidant activity of proanthocyanidins were 207.7mg and 156.36 mg. CONCLUSION: It was supported support that combined ABTS and FRAP methods, because of their conveniently, and their reliable results, especially in appropriately be used as the evaluation methods for antioxidant capacity of bioactive substance in vitro. PMID- 19548566 TI - [Action of silky chicken-Gastrodia elata Blume nutrient solution on immunoregulatory function in mice]. AB - OBJECTIVE: To study the immunoregulatory function of Silky Chicken-Gastrodia Elata Blume nutrient solution in mice. METHODS: Body weights, relative weights of immune organs, celluar immune functions, humoral immunity functions, mononuclear macrophage activitied and NK cell activities were tested in mice treated at the doses of 16.7, 33.4 and 50.0 ml/kg bw of Silky Chicken-Gastrodia Elata Blume nutrient solution. RESULTS: Silky Chicken-Gastrodia Elata Blume nutrient solution could remarkably increase the ratio of thymus to weight (P < 0.05) and the number of plague forming cells of mouse spleen cells (P < 0.01). NK cells activities were significantly increased (P < 0.05) under the conditions of twenty fold or thirty fold dosage of the the nutrient solution. CONCLUSION: It was suggested that Silky Chicken-Gastrodia Elata Blume nutrient solution could have strong strengthening function on immune system. PMID- 19548567 TI - [Effects of expression of fatty acid synthetase and phosphoenolpyruvate carboxykinase in offspring rats by high protein intake in pregnancy]. AB - OBJECTIVE: To investigate the effect of pregnant nutritional manipulation on offspring mRNA expression of fatty acid synthetase (FAS) and phosphoenolpyruvate carboxykinase (PEPCK). METHODS: Wistar pregnant rats fed on standard diet and high protein diet (HPD), respectively. The male offspring were divided into control and high protein group (HP) in accordance with the diets of their dams. Offspring were cross-fostered and fed on standard diet after weaning. At adult, some offspring randomly selected from control group, namely as high fat control (HFC), HFC and HP group rats fed on high-fat diet (HFD) to induce obesity. At different stages, tissue was collected for analyzing expression of FAS and PEPCK mRNA by fluorescent quantitation PCR. RESULTS: In HP, body weight and obesity rates were decreased after weaning and HFD (P < 0.05). The abundance of PEPCK mRNA was persistently decreased in HP, and the abundance of FAS mRNA in HP was down-regulated before and after HFD vs. control or HFC. CONCLUSION: The increase of protein intake during pregnancy could decrease the expression of FAS and PEPCK mRNA, and prevent offspring from HFD-induced obesity in adult. PMID- 19548568 TI - [Effects of alginate on Th1/Th2 cytokines in vivo and in vitro of mice spleen lymphocytes]. AB - OBJECTIVE: To study effects of alginate on Th1/Th2 cytokines in vivo and in vitro of mice spleen lymphocytes. METHODS: In vivo test: Fifteen C57BL/6 mice were randomly divided into one negative control group and two Alginate groups, which were respectively given purified water or 0.50, 1.50 g/kg bw alginate everyday. After 30 days, mice spleen lymphocytes were separated and cultured for 48h. Culture supernatant were gathered for analysis of IL-2, IL-4, IL-5, TNF and IFN gamma with cytometric bead array (CBA). In vitro test: C57BL/6 mice spleen lymphocytes were separated and respectively cultured with 25 mg/ml or 50 mg/ml alginate for 48h, and then culture supernatants were gathered for analysis of IL 2, IL-4, IL-5, TNF and IFN-gamma with cytometric bead array (CBA). RESULTS: In vivo test: Compared with the negative control, the IFN-gamma of 0.5 g/kg bw and 1.5 g/kg bw alginate groups showed marked elevation (P < 0.01), while IL-2 and IL 4 were both decreased (P < 0.05, P < 0.01). In vitro test: Compared with control group, TNF, IL-2 and IL-4 of 25 mg/ml and 50 mg/ml alginate groups (P < 0.05, P < 0.01) and IL-5 of 50 mg/ml alginate group (P < 0.01) were all increased. CONCLUSION: No matter in vivo or in vitro, alginate both can affect the Th1/Th2 cytokines of mice spleen lymphocytes. PMID- 19548569 TI - [Levels of mineral elements composition and heavy metal pollution in human breast milk in Shenzhen City]. AB - OBJECTIVE: To investigate the levels of some mineral elements composition and heavy metal in human breast milk in Shenzhen, and estimate the status of infant with breast feeding. METHODS: ICP-MS and ICP-AES instruments were used to detect the levels of four macroelements of Ca, K, Na, Mg and four microelements of Zn, Fe, Se, Cu, totally eight minerals as well as three heavy metal of Pb, As and Cd in human breast milk. 60 breast milk samples were collected from the women aged 20-35, lived in Shenzhen over 5 years, postdelivery 3 weeks to 2 months from Jul. to Nov. 2007. RESULTS: Average concentration for four kinds of macroelement of Ca, K, Na, Mg was 280.22, 498.61, 188.65 and 28.31 mg/L respectively, that for four kinds of microelements of Zn, Fe, Se, Cu was 2.29 mg/L, 358.88 microg/L, 8.28 microg/L and 339.16 microg/L respectively in 60 human breast milk samples. Average concentration of Pb was 2.13 microg/L in 60 human breest milk samples, and heavy metal As and Cd were non-detected. CONCLUSION: The nutrition status of four kinds of macroelement of Ca, K, Na, Mg and three kinds of microelements of Zn, Fe, Cu in 60 human breast milk samples were reasonable, but the lack of microelement Se and the pollution of the heavy metal should been taken into account. PMID- 19548570 TI - [A following up survey on cadmium level in rice in contaminated area, Jiangxi Province]. AB - OBJECTIVE: To observe rice cadmium level in a contaminated area where has being continuously polluted for 19 years since last investigation. METHODS: Samples of late rice (polished) grown in 2006 were collected from farmer household in contaminated and control area and its cadmium concentration were detected with IPC-MS. RESULTS: Samples of polished rice from the polluted and control areas were detected. The average cadmium concentration from last investigation area 19 years before was 0.59 mg/kg (0.05 mg/kg for rice from control area), it upraises about 30% corresponding to average of rice from the same areas detected before 19 years. The average cadmium level of rice from whole contaminated areas (including last investigation and expanding areas) was 0.26 mg/kg and it is greater than that in control areas (0.06 mg/kg) and the limited value of national hygiene standard (< 0.20 mg/kg). There was about 64%, 23.17% and 11% of samples have higher level than 0.20 mg/kg (the limited value of national hygiene standard), 0.4 mg/kg (the limited value of Codex General Standard) and 1.0 mg/kg (cadmium rice) respectively. There was a higher percentage (36.8%) of Cadmium-rice (greater than 1.0 mg/kg) in some heavy contamination villages. CONCLUSION: Environmental cadmium pollution has being existing for 19 years since last investigation. Average cadmium concentration of rice from whole country polluted areas was higher than that of national hygiene standard. Percentage of "cadmium rice" might be higher in some villages. PMID- 19548571 TI - [Study of validity and reproducibility of food frequency questionnaires for residents over 50-years-old in Xi'an City]. AB - OBJECTIVE: To examine the validity and reproducibility of Xi'an food-frequency questionnaire (FFQ) used to evaluate the diet intakes for residents in Xi'an City. METHODS: 125 males and females aged 50-80 years, from districts of urban Xi'an City, participated and completed all the six interviews in this study. Reproducibility of the questionnaire measurements was obtained by a repeated administration of the FFQ at one year interval. The mean of 24-hour dietary recalls (24-HDR) applied at three months intervals served as reference method for the estimation of the relative validity of questionnaire measurements. RESULTS: Spearman correlation coefficients for reproducibility between two FFQs ranged from 0.38 for cakes to 0.74 for tea. Coefficients for validity assessed by four 24-HDRs were statistically significant for almost all food groups ranged from 0.12 to 0.74. After correction for within-subjects error, all correlation coefficients were improved, ranged from 0.17 to 0.79. CONCLUSION: Therefore, Xi'an FFQ appears to be reasonably valid and reliable for most food groups. PMID- 19548573 TI - [Study on vitamin A nutrition status in plasma of different economic level of Chinese 3- 12-year-old children]. AB - OBJECTIVE: To know the Vitamin A nutrition status of different economic level of Chinese 3-12y children. METHODS: Blood samples were drown from 132 investigated counties of National Nutrition and Health Survey 2002, while each age group of children 6 blood samples were drown with boys and girls respectively. All blood samples were determined with HPLC. RESULTS: The vitamin A level in plasma is different in different economic level, and the vitamin A nutrition status is correlative with the family income. CONCLUSION: The result also indicates that the vitamin A level in plasma of 3-12y children is influent with the family economic level, and the result will provide the data to work out the strategy to improve the vitamin A nutrition status. PMID- 19548574 TI - [Study on nutritional status of primary and middle school students in Huai'an and Nanjing]. AB - OBJECTIVE: To study the nutritional status of primary and middle school students in Huai'an and Nanjing. To discuss the factors which affect the nutritional status of students and provide evidence for intervention among them. METHODS: Height, weight and hemoglobin indexes were measured among 2111 students in Huai'an and 1280 students in Nanjing. The body mass index and Rohrer index were used to assess the status of nutrition. WHO measures and standards were introduced to screen for the anemia ones. RESULTS: The prevalence rates of obesity, overweight, malnutrition and anemia were 9.8%, 3.6%, 17.6% and 25.3% respectively; while the prevalence rates of obesity, overweight, malnutrition and anemia were 13.8%, 8.4%, 23.2% and 22.4% in Nanjing. CONCLUSION: The height and weight of the students in Huaian were lower than the Jiangsu average levels of the students in Huai'an. PMID- 19548572 TI - [Study of establishing feeding index for children aged 6-23 months in rural China]. AB - OBJECTIVE: Infant and child feeding index was used to examine its associations with nutritional status of children aged 6-23 month in rural China. METHODS: Data come from China rural children nutrition surveillance in 2005, age-specific infant and child feeding index (ICFI) is developed based on breastfeeding and complementary feeding practices, and the associations between ICFI and weight-for age (WAZ), length-for-age Z scores (LAZ) were examined. RESULTS: The mean of ICFI was 8.61 +/- 2.56, ICFI was positively associated with LAZ in 6 - , 9- and 12 -23 month age group (P <0.01), ICFI was positively associated with WAZ in 9 - and 12 23 month age group (P < 0.05). Adjusted potentially confounding influences, ICFI remained positively association with both LAZ and WAZ (P < 0.01). Among the components of ICFI, dietary diversity, meal frequency and complementary feeding in a week were positively associated with LAZ (P < 0.01), breastfeeding, meal frequency and complementary feeding in a week were positively associated with WAZ (P < 0.05). CONCLUSION: ICFI could be used to assess effect of feeding practices on child growth in rural China. PMID- 19548575 TI - [Dietary intakes of mercury by children and adults from Jinhu area measured by the duplicate portion study]. AB - OBJECTIVE: To understand the dietary mercuric intakes of children and adults from Jinhu area and assess its safety. METHODS: The duplicate food portions of 62 subjects were collected over 3 consecutive days. Mercuric levels in duplicate samples were determined by the atomic fluorescence spectrometry. The safety of dietary mercuric intakes of children and adults from Jinhu area was evaluated by using the dietary mercuric PTWI recommended by JECFA of FAO/WHO. RESULTS: It was found that the percentile 97.5 weekly dietary mercuric (of PTWI) intakes of children and adults were 2.92 microg/kg bw (58.4%), 1.67 microg/kg bw (33.4%) respectively. The weekly dietary mercuric intakes of children calculated on a body weight basis were higher than that of adults ( z = 4.0406, P < 0.0001). CONCLUSION: Although the dietary mercuric intakes of children and adults are safe, it should be decreased to a lower level. Moreover, the dietary exposure to mercury are higher for children and should be considered seriously. PMID- 19548576 TI - [Influence of family factors on self-efficacy of the middle school students in Baise City]. AB - OBJECTIVE: To understand the relationship of self-efficacy and family factors of the middle school students in western of Guangxi. METHODS: A total of 778 Zhuang students selected by stratified cluster sampling were questionnaired with the general perceived self-efficacy scale, parents' education scale and Family Environment Scale. RESULTS: (1) Except of education of mother and parents' marriage (15.107/0.002, 6.018/0.049), there were no significant differences for other demography variables. (2) There was significant positive correlation between self-efficacy with cohesion, emotion expression, emotion warm and understanding, but negatively with conflict. (3) The multiple linear regression showed that self-efficacy was affected by cohesion, conflict, emotion warm and understanding, refusing and denying, over-protection and over-interference, as well as education of mother (F = 87.812, R2 = 0.558). CONCLUSION: There was close relationship among self-efficacy, family environment and parents education. It was helpful to establish favorable self-efficacy in scientific education style and warm and understanding of family environment. PMID- 19548577 TI - [Primary and secondary school counseling staff self-efficacy relevant factors in Foshan City]. AB - OBJECTIVE: To understand the general self-efficacy of primary and secondary school counseling staff in Foshan City. METHODS: Using multi-stage random sampling method to choose 108 counseling staff from primary and secondary school in Foshan City. Taking a self-made questionnaire, General Self-Efficacy Scale (GSES) and Simplified Coping Style Questionnaire (SCSQ) for investigation. RESULTS: The self-efficacy of the Primary and secondary school counseling full time and part-time staff have significant difference (P = 0.003), the self efficacy of full-time staff above than part-time staff; The negative coping style of the Primary and secondary school counseling full-time and part-time staff have significant difference (P = 0.007), the negative coping style of part-time staff above than full-time staff; different academic counseling staff of the positive coping style have significant difference (P = 0.039), the positive response of Master's degree and above is highest scores, college staff and below is lowest score. CONCLUSION: The self-efficacy of Primary and secondary school counseling staff is related to positive coping styles and the nature of work (full-time or part-time). PMID- 19548578 TI - [Study of ALAD and VDR gene polymorphisms associated with lead nephrotoxicity susceptibility]. AB - OBJECTIVE: To explore lead nephrotoxicity associated with delta-aminolevulinic acid dehydratase, vitamin D receptor gene polymorphism as well as joint effect of gene-gene, gene-environment. METHODS: 233 occupational lead-exposed workers were involved in this study. Then blood lead was detected as lead exposure biomarker. Urinary N-acetyl-beta-D-glucosamindase and beta2-microglobin of those workers were measured as biomarkers of nephrotoxicity. PCR-RFLP method was used for gene polymorphism analysing. RESULTS: Lead exposure concentration was higher than national limit, NAG acticity were (2.12 +/- 0.07) U/mmol Cr in workers with ALAD1 2/ALAD2-2 genotype and (1.73 +/- 0.03) U/mmol Cr in workers with ALAD1-1 genotype (P < 0.05). In the same condition, beta2-MG were (20.94 +/- 1.12) microg/mmol Cr in workers with VDR-Bb genotype and (15.28 +/- 0.09) microg/mmol Cr in workers with VDR-bb genotype (P = 0.01). Analysis of logistic regression confirmed that lead exposure and high blood lead as well as combined effect of gene-environment were responsible for lead nephrotoxicity in workers (OR = 6.58, 2.41, 3.01). CONCLUSION: Polymorphisms of ALAD and VDR gene may play an important role in lead nephrotoxicity in high lead-exposed workers. Joint effect of gene-environment could be involved in lead nephrotoxicity in workers. PMID- 19548579 TI - [Analysis of quality of life and its influence factors of urban residents in Chongqing City]. AB - OBJECTIVE: To explore the quality of life and its influence factor in urban residents in Chongqing city. METHODS: By stratified cluster sampling, 738 urban residents in Yuzhong and Jiulongpo Districts were collected and were assessed with the version 2 of SF-36 and the general characteristics. RESULTS: Quality of life in the eight dimensions, physical health (PF) and social function (SF) dimension scored higher, overall general health (GH) got the lowest score in Chongqing residents. The work and rest orderliness, appetite, sleep, chronic disease, job satisfaction, and family, interpersonal relationships could affect every dimensions on the quality of life by single-factor analysis. Gender, age, education, occupation, income and other factors could also influence qualities of life in different levels. Coping trend, chronic disease, doss, income, the satisfactory of life, interpersonal relationship, and the work and rest orderliness could be were main influence factors in qualities of life. CONCLUSION: The prevention of chronic diseases, economic development, and adoption of healthy lifestyle could promote improvement of the quality of life in urban residents in Chongqing City. PMID- 19548580 TI - [Immune function changes caused by contact dermatitis in mice after intracutaneous injection of trichloroethylene]. AB - OBJECTIVE: To investigate the contact dermatitis of BALB/c mice exposed to Trichloroethylene (TCE) via back intradermal injection and observe the influence on immune function of BALB/c mice. METHODS: BALB/c mice were exposed to TCE through intradermal injection. The organ coefficient of spleen and thymus were calculated. Lymphocytes viability and lymphocytes proliferation assay, Macrophage activation and NK cell activity assay were carried out; the content of TNF-alpha and IFN-gamma in the serum were measured. RESULTS: The thymic organ coefficient of TCE group was significantly lower than that of the negative control group (P < 0.01). The lymphocytes viability and lymphocytes proliferation of TCE group were obviously higher than those of the negative control group (P < 0.01). Compared to negative control group, the response of macrophage activation and NK cell activity of the TCE group were enhanced markedly (P < 0.01). In TCE group the serous content of TNF-alpha was evidently higher than that of the negative control group (P < 0.01). CONCLUSION: Trichloroethylene as an allergen can cause contact dermatitis whose mechanism was supposed to be a delayed-type hypersensitivity. PMID- 19548581 TI - [Relationship of PAHs levels in umbilical cord blood and the PAHs exposure between the mother and the paired newborns]. AB - OBJECTIVE: To determinate the present polycyclic aromatic hydrocarbons (PAHs) levels in cord blood in order to discuss the PAHs exposure relationships between mother and paired newborns. METHODS: 347 pregnant women joined the study and the information of the 271 paired mother-newborns were used to analysis the exposure relationship. Questionnaire and bio-samples were got during the period of October 2006 to January 2008. High-performance liquid chromatography (HPLC) was used to determine the PAHs (7 kinds) levels in cord blood and maternal blood. RESULTS: In the 271 paired mother/newborns, several kinds of PAHs were detected in nearly all the serum of the subjects. The serum concentrations of B(k)F, B(a)P and DB(a, h) A in cord blood were significantly higher than those in paired maternal blood. In addition, the serum concentrations of B(b)F in cord blood was higher than that in maternal blood (P > 0.05). CONCLUSION: There are several kinds of PAHs detected in the umbilical cord blood and the PAHs levels in cord blood is the same as or even higher than that in the maternal blood, which means that could be is very important to develop the prenatal exposure assessment. PMID- 19548582 TI - [Effects of lanthanum on memory and expression of c-fos mRNA and c-Fos protein of cerebral cortex in rats]. AB - OBJECTIVE: To investigate the effects of lanthanum on memory, and explore the corresponding mechanism from immediate early gene expression of cerebral cortex in rats. METHODS: Female adult rats were divided into control, low, middle, and high dose lanthanum groups at random. Offspring of lanthanum groups were administrated with lanthanum from birth to weaning for one month. Ability of memory of offspring was detected by neuroethology testing. The expression levels of c-fos mRNA, c-Fos protein and Nissl body in cerebral cortex of offspring were also measured. RESULTS: The data in neuroethology testing showed that memory of offspring in lanthanum groups were significantly lower than control level and presented dose-effect relationship to some extent. The expression levels of c-fos mRNA and c-Fos protein in cerebral cortex of low dose lanthanum group were lower significantly than control values. As to middle dose lanthanum group, the expression levels of c-fos mRNA in cerebral cortex were lower significantly than control values, and c-Fos protein expression were lower significantly than control and low dose lanthanum group. Both c-fos mRNA and c-Fos protein expression in cerebral cortex of high dose lanthanum group decreased significantly as compared with control and low dose lanthanum group. In addition, the expression levels of Nissl body in cerebral cortex of low dose lanthanum group were lower significantly than control values. The expression levels of Nissl body in cerebral cortex of middle and high dose lanthanum group decreased further. CONCLUSION: Lanthanum induced deficit in memory, which is possibly related to decreased levels of immediate early gene expression and neuronal function in cerebral cortex caused by lanthanum. PMID- 19548583 TI - [Effect of hydroquinone on the expression of ubiquitin-conjugating enzyme Rad6B in human L-02 hepatic cells]. AB - OBJECTIVE: To study the effects of hydroquinone (HQ) on expression of ubiquitin conjugating enzyme Rad6B in human L-02 hepatic cells. METHODS: After L-02 hepatic cells were at the concentrations of 0, 5, 10, 20, 40, 80, and 160 micromol/L HQ for 24h, and the levels of Rad6B mRNA and protein expressions in L-02 hepatic cells were detected by real-time fluorescent quantitative polymerase chain reaction (QPCR) technique and Western blot method respectively. RESULTS: It was found that HQ in the range of 0-160 micromol/L could induce increases in the expressions of Rad6B mRNA and protein, which was in a dose-dependent manner. The levels of Rad6B mRNA and protein expressions in the treated group of 5, 10, 20, 40, 80, and 160 micromol/L were higher than those in the control (P < 0.01), levels of Rad6B mRNA and protein expressions reached the maximun when L-02 hepatic cells were treated by HQ at the concentrations of 160 micromol/L, and the relative quantity values were 4.35 and 0.85, respectively. CONCLUSION: HQ could regulate up the expression of Rad6B in L-02 hepatic cells. PMID- 19548584 TI - [Effect of t10, c12-conjugated linoleic acid on fatty acid metabolism in C2C12 myotubes]. AB - OBJECTIVE: To study the effect of t10, c12-conjugated linoleic acid (t10, c12 CLA) on fatty acid metabolism in myotubes under the high free fatty acid condition. METHODS: We used palmitate to simulate a high free fatty acid condition for C2C12 myotubes in vitro and administered t10, c12-CLA across the groups. The expression and translocation of FAT/CD36 were determined by fluorimetric immunifaction using laser scanning confocal microscope. A series of enzymes related to fatty acid metabolism, together with AMP-activated protein kinase (AMPK), were determined by western blot. RESULTS: t10, c12-CLA dose dependently increased the translocation of FAT/CD36 to cellular membranes, and increased the expression of p-ACC and CPT-1 in myotubes under the high free fatty acid condition. Moreover, t10, c12-CLA activated the AMPK in myotubes. CONCLUSION: CLA can increased the fatty acid oxidation in myotubes via activating AMPK. PMID- 19548585 TI - [In vitro regression model of glycemic index for carbohydrate-riched foods]. AB - OBJECTIVE: To Establish the in vitro regression model of glycemic index for carbohydrate-riched food by the in vitro digestibility and composition characteristics. METHODS: Thirty products were commercially available and selected on the basis of their high carbohydrate content. After determined fat, protein and carbohydrate constitutes (sugar, starch), an in vitro method for digestibility characteristics were developed to measure hydrolyzed starch at 20 min, 120 min, 240 min, 16 h, and non-digested resistant starch (RS). In vivo glycemic responses were determined by standardized methods. The relationship between the compositions and GI were also discussed through stepwise regression methods. RESULTS: The observed GI ranged from 26 to 113, and correlated strongly with the digestibility profile of carbohydrates. Significantly positive correlation of S20, S120 (P < 0.05) and negative correlation of RS (P < 0.01) to GI was observed. Easily available glucose (EAG, sum of content of S120 and glucose) and RS were selected as variables to setup regression equation for in vitro GI prediction, GI = 39.65 + 1.008EAG-1.072RS. CONCLUSION: The findings indicate that the digestibility profile of carbohydrates is very essential to explain in vivo glycemic response of carbohydrate rich foods and predict GI value. PMID- 19548587 TI - [Growth characteristics and metabolomic sampling strategy of Lactobacillus sp. 30a(ATCC33222)]. AB - OBJECTIVE: Sampling strategy for metabolomic studies of ATCC33222 was discussed according to its grown patterns under six different media. METHODS: Six media with different ingredients or pH value were prepared and ATCC33222 strain cultures were harvested from each medium. Culture samples were collected every 1 4 hours and OD600 and pH values were determined thereafter. Growth curves and pH curves were constructed according to these data. RESULTS: Growth curves were different between ATCC33222 strain cultures. MRS + aa pH 3.5 and MRS + aa + COS media showed longer lag phase, medium with additional glucose had the longest log phase, MRS + aa under pH 6.5 and 5.0 media showed shorter stationary phase. MRS, MRS + aa + glucose and MRS + aa + COS had similar pH curves while the other three pH curves were different from each other. CONCLUSIONS: ATCC33222 strain may have the ability of dynamic metabolic'regulation to accommodate different culture environment or energy source. In metabolomic studies, pH 3.5 and pH 6.5 were recommend for culture condition. Amino acids could be used as ingredient in MRS while glucose was only optional, pH 5.4-5.9 could be important for lactic acid bacteria metabolomic studies. PMID- 19548586 TI - [End product enrichment changes of tracer leucine under the status of infusion and analysis on 13CO2 production rate]. AB - OBJECTIVE: To analyze the end product enrichment of the stable isotope labeled amino acid in different time points and the level of catabolism of different protein dose groups under the infusion state of the labeled amino acid in the study of amino acid metabolic kinetics, supplying the data support for the the setting of sampling time in the study of the physiological requirement of protein and amino acid. METHOD: Twenty subjects were divided into two groups and were supplied two different doses of protein, all subjects were administered the 13C leucine for 3 hours by the left basilic vein at 12:00 on day 6 and respired air samples were collected per 15 minutes during the last 30 minutes of infusion, analyzing the 13C enrichment difference at three time points and the 13CO2 production rate of the two dietary protein dose groups to determine whether 13C enrichment was stable or not and how the dietary protein dose influence the 13CO2 production rate. RESULT: The difference of 13C enrichment is not significant at three time points for every group subjects (P > 0.05), the 13CO2 production rate of the protein dose group of 0.89 g/kg/d was significantly higher than that of the protein dose group of 0.89 g/kg/d (P < 0.05). CONCLUSION: It was suggested that determine one specific time point were determined to compare the metabolic kinetics changes of the different protein and amino acid dose groups, when the physiological requirement of protein and amino acid was explored, and the catabolism level in the bigger dose group was higher than that in the smaller dose group. PMID- 19548588 TI - [Comparison of lead concentration and isotope ratios in children's blood and hair]. AB - OBJECTIVE: To explore the characters of lead isotope ratios in children blood and hair, in order to obtain new data of the cumulated lead sources of human body. METHODS: 28 children in Beijing area were randomly sampled to take blood and hair with a 1:1 sex ratio, half of them from the city and the others from the village. The lead concentrations and isotope ratios in the samples were then determined by ICP-MS. RESULTS: Lead isotope ratios in blood had significant differences between the children living in the city and those in the village, while no lead isotope ratio differences in hair could be found between the children living in the two area. The lead isotope ratios in children blood were significantly different from those in children hair in both area, in the other words, the lead fingerprints were different between blood and hair. CONCLUSION: (1) The concentrations of lead pollution of children were resemble between the city and village in Beijing, but the lead sources in the city and village might be different from each other. (2) Organs or tissues in human body might have its own mechanism to accumulate the lead isotopes. (3) It was necessary to study if the lead in hair could be used as a biomarker of lead source. PMID- 19548589 TI - [Progression of RNA interference on pathogenesis and treatment of neurodegenerative diseases]. AB - Neurodegenerative diseases could be suitable for RNA interference (RNAi), and complexity of their pathogenesis could make into RNAi as a strong research means, and in this article the application of RNA interference in pathogenesis and treatment of neurodegenerative diseases were reviewed. PMID- 19548590 TI - Metabolic syndrome (does it have a common denominator?). AB - The insufficient or inappropriate supply of PUFA OMEGA-3 (esp. docosahexaenoic and eicosa pentaenoic acid DHA and EPA) can be suggested to be a common factor of the four metabolic syndrome's clinical manifestations. DHA can be considered as one of the most important element of the membrane. The protective and beneficial effects of DHA (and EPA) result from the dynamic qualities of its molecular structure as well as from its recently detected effects on the genetic expression of several cytokines, enzymes etc. Dietary supplementation of DHA improves all clinical symptoms and laboratory biochemical markers of the metabolic syndrome. Together with that, depletion of DHA was found in diabetes and in several cardiovascular diseases. Also the developmental aspect and approach supports our view. With high probability DHA represents one of the connecting components of the development of metabolic syndrome's clinical manifestations. Metabolic syndrome could be therefore interpreted as an insufficient function of the cellular membrane. PMID- 19548591 TI - Minireview: molecular structure and dynamics of drug targets. AB - Summary of lectures presented at the Czech and Slovak Pharmacological Meeting, Prague, September 2008. PMID- 19548592 TI - Genomic polymorphism and sepsis--is there a reason for optimism? AB - There is no doubt that, in infectious disease, genetic predisposition plays a very important role in clinical outcome. Sepsis is a polygenic syndrome initiated by infection. A fact confounding the situation is that two factors--the macroorganism and the microorganism--are at play at the same time; hence of genotype effect must be assessed in light of their interaction. From a phylogenetic point of view, infectious disease is a companion of man throughout their life and its role in terms of function of the system of innate immunity is perceived as a beneficial one. However, the presence of a major antigen load by the infectious agent results in pathological responses at the levels of the macroorganism. Assessment of the severity of the inflammatory process on the basis of genetic predisposition is a most challenging issue. Genetic polymorphisms in the immune response to infection have been shown to be associated with clinical outcomes. The advancement of single nucleotide polymorphism (SNP) genotyping in basic genes--CD14, Toll like receptors, LBP, cytokines, cytokine receptors and coagulation factors have provided valuable information on the interaction of the macro and microorganisms. The understanding of the variation in genes and differences in response to infection may contribute to tailored diagnostic and therapeutic interventions with improved outcome in these patients. PMID- 19548593 TI - Comparisin of low energy breakfast based on special egg white spread product with a standard breakfast. AB - AIM: The aim of the study was to evaluate the influence of a new form of low calorie breakfast in comparison to standard low fat cheese breakfast. A special spread based on egg white was used (Energy content per 100 g: 319 kJ versus 802 kJ, fat content 1.15 g versus 15 g). Egg white derived products are widely used for weight reduction in Czech Republic. METHODS: 12 non-diabetic patients were included into the study (7 men and 5 women), mean age 38.5+/-10.7 years, and mean BMI 24.6 kg/m2. Standard breakfast was served first in a time interval of 1 week (Krajanka cheese) and after a week wash-out period low energy breakfast based on egg white was served for the same time. Glucose level, insulinaemia and C peptide were evaluated in 0, 60, 120 minute after breakfast. Psychological questionnaire was used to evaluate the taste and satiety. RESULTS: Standard breakfast: insulinaemia 6.3...34.6...11.0 IU, C peptide 0.6...1.6...1.0 pmol/l, glucose 5.0...5.4...4.8 mmol/I Low energy breakfast: insulinaemia 6.6...24.6...10.7 IU, C peptide 0.6...1.4...1.0 pmol/I, glucose 4.8...5.1...5.0 mmol/l. Paired t-test was used for evaluation. No difference in blood glucose level was found, borderline significances of insulinaemia in 60 minute (p= 0.056) and in C peptide (p= 0.089). Significant difference in decrease of insulinaemia between 60 and 120 minute (p=0.03) and borderline significant decrease in C peptide (p=0.055) shows shorter insulin secretion after low calorie breakfast and good insulin sensitivity in this group of lean subjects. Higher satiety and less-worthier taste were found using low energy breakfast. CONCLUSION: Low calorie breakfast induces lower and shorter insulin secretion and has a good metabolic effect in lean subjects. Testing in obese subjects will follow. Egg white derived low calorie products have a beneficial effect on insulin response without any difference in ingested carbohydrate quantity. PMID- 19548594 TI - Stability of free beta-hCG in the routine screening of Down syndrome in the first trimester of pregnancy. AB - Stability of free beta-human chorionic gonadotropin in maternal blood upon storage at ambient and refrigerator temperatures as well as stability under conditions simulating postal transport were studied. A group of 26 women in the first trimester of pregnancy were included in the study, blood samples were taken during the routine check-up. Blood samples were aliquoted stored in different conditions that should mimic the transportation and then analysed for free beta hCG. No significant differences were found in the free beta-hCG levels in samples after 24 and 48 hours resp., stored in refrigerator (2-8 degrees C) without separation. In samples stored in laboratory temperature without separation the average concentration increased from 11 to 20%. Six blood samples were stored at 35 degrees C for 5 hours and then in laboratory temperature. In this group the average increase of results was from 14.3% to 132.2%. Separation of the sera for Down syndrome screening in 4 hours after withdrawal is necessary. Cooling during any storage, including transportation is highly recommended as the preanalytical phase has a high impact for the analysis. PMID- 19548595 TI - Influence of hydroxyapatite crystallinity on the growth of keratinocytes. AB - Dental implantology is a field, which has made a great progression recently. The main task nowadays is to shorten the healing period and so improve the comfort for the patients. One possibility how to full fil this task is to coat the surface of the implant. Very promising material seems to be hydroxyapatite, which is a natural component of human body and suitable method is the pulsed laser deposition. In our study we tried to evaluate difference between crystalline and amorphous hydroxyapatite coated dental implants from the biological point of view. We found that the cells were able to adhere to all of our studied samples. The worst proliferation of fibroblasts was found on the amorphous coating, whereas the adhesion was fully comparable with other surfaces. The level of keratinocyte differentiation was the same on both of the studied surfaces. PMID- 19548597 TI - Advanced oxidation protein products in obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is a risk factor of hypertension, coronary artery disease and stroke. OSA is also considered a cause of accelerated atherogenesis. Advanced oxidation protein products (AOPP) are among the biochemical indicators of higher risk of atherogenesis as an independent risk factor for coronary artery disease. 20 men suffering from OSA were examined using night polygraphy, the AOPP were determined from their morning blood samples. The mean AOPP concentration in the patients group was 91.8 (SD=42.3) micromol/l, in the control group 76.2 (SD=35.3) pmol/l, the difference was not significant. The AOPP were found correlated with the AHI (apnoe/hypopnoe index) (R=0.485, P=0.030). The results support the hypothesis that OSA increases the oxidative stress and atherogenesis. PMID- 19548596 TI - Quantitative analysis of procalcitonin after pulmonary endarterectomy in relation to cytokines and C-reactive protein. AB - BACKGROUND: Endarterectomy of the pulmonary artery (PEA) is a potential curative treatment method for selected patients with chronic thromboembolic pulmonary hypertension (CTEPH). The postoperative course after PEA is associated with high rate of early postoperative mortality, which ranges from 5 to 23%. The aim of this prospective clinical study was to assess the time course of procalcitonin (PCT) after uncomplicated PEA in relation to other inflammatory parameters. PATIENTS AND METHODS: 32 patients with CTEPH treated with PEA using cardiopulmonary bypass and deep hypothermic circulatory arrest were included into study. PCT, C-reactive protein (CRP), interleukin (IL)-1beta, IL-6, IL-8, and sTNFRI arterial concentrations were measured before/after sternotomy, last DHCA, separation from bypass, and repeatedly to 48 h after sternotomy. RESULTS: Mean duration of CPB was 338.2 min.; mean circulatory arrest time 39.9 min. The initial decline of PCT, IL-6, and IL-8 was followed by an increase culminated 6 24 h after sternotomy. PCT peak was detected +24 h (1.97 ng/ml, 1.70-2.54). IL-6 culminated after separation from CPB, IL-8 was highest 12h after sternotomy. PCT levels correlated with IL-6 (r = 0.81), CRP (r = 0.72), and sTNFRI (r = 0.58). CONCLUSION: Postoperative PCT culmination was delayed in alignment to pro inflammatory mediators as IL-6 and IL-8. Positive correlation between PCT and IL 6 corresponded with the role of IL-6 in PCT induction in perioperative phase. Plasma PCT estimation extended to a measurement of CRP and IL-6 may become a useful complementary examination in early postoperative period after PEA. PMID- 19548598 TI - Uterine arteries doppler velocimetry provides 3-years follow up endometrial ablation outcome. AB - The aim of this study was to assess whether uterine artery Doppler velocimetry [pulsatility index (PI) and resistance index (RI)] and thickness of the endometrium (TE) are able to predict 3-year clinical outcome after endometrial ablation (EA) for dysfunctional uterine bleeding (DUB). This was a prospective, observational study of 29 women of whom 22 were amenorrhoeic (A) and 7 eumenorrhoeic (E) at the end of the first postoperative year. The PI, RI and TE were measured prior to and 1, 6 and 12 months after EA. Statistical analyses were performed using BMDP statistical software, discriminant analysis, ANOVA and T test. Using the calculated classification function (CF) with the three parameters PI, RI and TE measured 12 months after FEAT, we were able to accurately (100%) specify which of the women will have A or E in 3 years. The predictive value of PI, RI and TE has been confirmed clinically in a minimum 3-year follow-up of outcome (ranging from 36 to 72 months [mean 55]). All A and E women have stayed in the same group (A or E) during the minimum of 3 years. In conclusion we found that PI, RI and TE measured prior to EA cannot predict the outcome, however these measurements performed 1 year after FEAT can predict the duration of A or E in the 3-year follow up. PMID- 19548599 TI - Internet supported standardized patient information for selfcare. AB - BACKGROUND: An informed patient is a better partner for the physician than an uninformed one. Such a patient will be more likely to practice effective selfcare, appropriately utilize the health care system, and be prepared to seek and obtain health care services that meet his needs. Objectives. To develop a system of presenting standardized medical information for the general population based on guidelines developed for general practitioners (GPs). METHODS: Guidelines are instruments to be used by GPs to assure comprehensive and consistent enhancement of the quality of care. At the same time, guidelines describe the actions which could be expected by patients in various clinical situations. These materials establish a good basis for the patient's understanding of health care procedures and when and how to utilize health services. These materials will have to be adapted in form, content, and nomenclature in order to be understandable with the background knowledge of the average "citizen". This information for patients is currently being developed in various forms of presentations and applications. The use of web-oriented tools is a priority, because web content offers the patient a very simple and easy way of browsing and searching for the latest relevant information on both basic and advanced/detailed levels, including supporting pictures and animations. Materials are also being developed in other formats such as printed leaflets, audio presentations and videoclips. RESULTS: Web content and printed materials based on clinical guidelines have been created. Focus groups of health professionals and non-professionals were used for evaluation of the materials. General practitioners have tested written materials in practices. CONCLUSIONS: The development and preparation of information and materials for patients has become an integral part of the GP practical guideline development cycle. PMID- 19548600 TI - Prolonged impairment of polymorphonuclear cells functions in one infant with transient zinc deficiency: a case report. AB - BACKGROUND: Zinc is an essential trace element for the immune system. The zinc deficiency diminishes antibody- and cell-mediated responses in man. Lymphopenia and thymic atrophy are usually the early hallmarks of zinc deficiency. Surprisingly, only scarce data are available about polymorphonuclear cells (PMNs) functions in infants with zinc deficiency. We present the results of immunological analyses in one infant with transient zinc deficiency due to decreased zinc concentration in mother milk resulting in severe lactogenic acrodermatitis enteropathica. MATERIAL/METHODS: Nine repeated examination of oxidative burst of PMNs and immunoglobulin levels using nitroblue tetrazolium dye test, chemiluminescence, flow cytometry and nephelometry were performed in the infant with severe zinc deficiency during 28 months period. RESULTS: The unusual prolonged but transient impairment of PMNs respiratory burst accompanied with hypogammaglobulinaemia developed since the age of 2.5 months. Dramatic improvement of the skin was observed within days with total resolution of skin lesions on the 9th day of zinc therapy, but decreased PMNs respiratory burst persisted until the age of 23 months. CONCLUSIONS: We conclude that zinc deficiency may lead to prolonged impairment of polymorphonuclear cells functions and hypogammaglobulinaemia. PMID- 19548601 TI - Unrecognized primary genital herpes infection (case report). AB - The authors present a case report of unrecognized herpes genitalis, which was caused by primary HSV-1 infection. The 28-year old female was examined by three board certified gynaecologist and initially treated as mycotic vulvitis. The authors point out the atypical course in the patient without anti-HSV-1 and HSV-2 antibodies. This infection is rather very painful, extensive and with complications. Thus, it is necessary to consider the diagnosis and to begin antiviral therapy as soon as possible. PMID- 19548602 TI - Ectopic parathyroid adenoma in child. AB - A 10-year old girl presented with fatigue, hypercalcemia, and subperiosteal phalangeal osteolytic lesions. Ultrasonography and MIBI scintigraphy showed a structure near the lower pole of thyroid gland. The structure macroscopically appeared as adenoma, histologically it was thymic tissue. Bilateral neck exploration together with exploration of cervical thymic extensions was performed; adenoma was not found. During next two years, the level of calcium and parathormone raised, bone mineral density decreased. Ultrasonography, MRI, CT and PET/CT were negative. Adenoma was located by MIBI-SPECT/CT near the left border of jugulum. It was found dorsolateral to left common carotid artery and removed. PMID- 19548603 TI - The life-work of Prof. MUDr. Zdenek Lojda, Dr.Sc., Dr. Med. et Iur. h.c. AB - Professor Zdenek Lojda, MD., Dr.Sc., Dr. Med.h.c., Dr. Jur. h.c., vice-rector emeritus of the Charles University and director emeritus of the Institute of Histology and Embryology of the 1st Medical Faculty of the Charles University in Prague died on the 24th April 2004. Born in Trebic (in a local maternal hospital on the 7th December 1927) he grew up in Moravske Budejovice where he graduated from a primary and secondary school (he passed out each class as well as the leaving exam at the local gymnasium in 1946 cum laude). He was growing up in an inspiring atmosphere of a family of a professor at gymnasium that helped him to form his fundamental moral attitude and to gain high knowledge of an almost Renaissance comprehension. He was a gifted linguist (he spoke fluently several languages including Latin) and musician (he did community singing among others) and he was interested in natural sciences. In 1946 he enrolled for the medical faculty and coincidentally he was grouped among students that completed histology and embryology at the Institute of Embryology headed by professor Zdenek Frankenberger, MD., who had recognised his interest in this field of study. The young student gave himself and his free time to the work of an unqualified lab worker and only later he became assistant conducting practical tutorials. He took his degree cum laude in 1952 and he received certificate of competence because of his excellent results. He showed his preoccupation with histology (strongly influenced by the personality of professor Frankenberger) after the graduation when he entered the job of the assistant professor at the Institute of Embryology and he stayed there until 1961. When he was about to decide which problems he should target, professor Frankenberger drew his attention to a very interesting part of histology that was just in advance and that was the use of diazonic salts for formation of colour reaction product demonstrating enzyme activity in tissues. In 1962 professor Frankenberger retired and Lojda left the Institute of Embryology for the 4th internal clinic to work in the Laboratory of Angiology housed at the 1st (Hlava) Institute of Pathologic Anatomy. This step influenced Lojda's professional occupations: in the Laboratory of Angiology he studied enzymes of blood vessel wall; the problems in gastroenterology solved at the 4th internal clinic and teaching hospital brought him to issues connected with enzyme garniture of the intestinal mucosal epithelium; his engagement at the Hlava Institute where he worked as well as house officer and later as pathology teacher caused that he subordinated his interest in enzyme garniture of various tissues to study and explanation of pathological processes using functional morphology. In years 1966 to 1975 he earned his master degree in pathology, he habilitated in pathology and was promoted to a professorship in pathology. PMID- 19548604 TI - EDs not taking chances with H1N1, protocols updated, supplies checked. AB - The ultimate severity of the current H1N1 outbreak is far from certain, but ED managers and other infection control experts are taking a proactive approach to ensure they are prepared for the worst. Here are some steps they're taking: Patients are being asked additional questions in triage to determine risk levels. Staff members are double-checking the fit on their N95 masks. Arriving EMS patients with suspected respiratory infections are remaining in the ambulance until ED determines where they should be taken. PMID- 19548605 TI - 'Reverse triage' adds to surge capacity. AB - Providing adequate surge capacity during a disaster is one of the greatest challenges of emergency response. Now, researchers have proposed a new process called "reverse triage" to help create surge capacity that otherwise would not exist. Patients who have only a slight chance of experiencing an adverse event within four days of leaving the hospital may be discharged to free bed space. ED staff can provide a daily initial reverse triage score for patients being admitted, even if a disaster is not imminent. While general guidelines can have great value, take the interests of the patient and their family into account when making discharge decisions. PMID- 19548606 TI - Staffing changes save ED more than $200K a year. AB - The ED at Dameron Hospital in Stockton, CA, estimates it has saved more than $200,000 a year simply by adjusting the way its staff is allocated. In addition, its rate of left without treatment was in the teens and is now 2%. Here are a few of their winning strategies: "Atypical" shifts were created to match the change in patient load at different times of the day and night. Every nurse was rated according to skill level, so that each shift now has the same balance of skill on its team. A computer portal was created to allow nurses to learn of shift openings. PMID- 19548607 TI - ED/hospitalist plan improves throughput. AB - An ED leader and the head hospitalist at Johns Hopkins Bayview Medical Center, Baltimore, MD, jointly developed a new plan for admitting patients from the ED that has decreased ED throughput for admitted patients 98 minutes (from 458 minutes to 360 minutes) from the same period a year earlier, despite an 8.8% increase in the ED census. Here are some of the keys: A hospitalist, with the treating ED physician, makes the final decisions for admitting ED patients to the cardiac ICU; the medical ICU; and the cardiology, pulmonary, and general medicine units. The rotating position, covered 24/7, also has responsibility for bed management. Objections of specialists in other departments were overcome by giving them input. PMID- 19548608 TI - Hospitals advised to play it safe when it comes to H1N1. AB - Review, drill on your existing response plans and look for any "holes" that may exist. Survey your staff to determine if you will have adequate coverage in the event of a pandemic. Make sure supplies are adequate and staff education is up to date. PMID- 19548609 TI - NQF endorses practices for safe lab medicine. AB - Specimen mix-ups can cause errors even if the tests were performed properly. More standardized approach can improve quality and safety. Lab leadership is a key component in addressing processes. PMID- 19548610 TI - Hospital discharge process can be more efficient. AB - A poorly run discharge process can result in nursing dissatisfaction, swing between high bed vacancies to crowding, and wasteful resource spending. Hospital examines nurse-patient ratios and makes changes that lead to lower turnover rates. "Holy Grail" of discharge planning process is measuring the time from when the physician writes an order to the time the patient is accepted on the floor. PMID- 19548611 TI - Scale measures quality of hospital discharge process. PMID- 19548612 TI - TJC makes it clear: get surgical smoke out of OR. PMID- 19548613 TI - [Structural-functional transformation of the malate dehydrogenase system of the bacterium Sphaerotilus sp. strain D-507 depending on nutritional mode]. AB - High-purity preparations of malate dehydrogenase (EC 1.1.1.37) were obtained by multistage purification from the bacterium Sphaerotilus sp. strain D-507 growing under different conditions. Under organotrophic conditions, the enzyme was dimeric; under mixotrophic conditions, dimeric and trimeric. On the basis of studied properties of the enzyme preparations, data on the activity of enzymes of the glyoxylate and tricarboxylic-acid cycles, and analysis of published data, it can be concluded that malate dehydrogenase isoforms are implicated in the adaptive response of bacteria to changing culturing conditions. PMID- 19548614 TI - [Analysis of genetic variation and differentiation in the pygmy wood mouse Sylvaemus uralensis (Rodentia, Muridae) aided by RAPD-PCR method]. AB - The present work ascertains distinct differentiation of the pygmy wood mouse into two groups of populations, which correspond to the European and Asian races, but can not, however, be considered valid species due to the low values of genetic distances between them. Neither of the used statistical methods explicitly verifies genetic isolation of the East-European and South-European chromosomal forms, which are close to one another and together constitute the European race. However, polymorphism of these chromosomal forms/races of the pygmy wood mouse differs each of them from the others, that can be explained both by the history of their formation (the degree of proximity to the ancestral population complex, isolation, etc.) and by unequal current environmental conditions. PMID- 19548615 TI - [Micromorphometric analysis of root cap cells in Allium cepa]. AB - A two-dimensional micromorphometric analysis of a root cap in Allium cepa included the measurement of the areas of about six thousand cell profiles on both longitudinal and transversal sections. Basing on the results of this analysis, quantitative descriptions of two cell populations from the columella and periferic part of the root cap have been created. The scheme of arrangement of root cap cells, located at a distance of 250 microm or less from a root tip, has been developed. PMID- 19548616 TI - [The influence of mineral nitrogen on legume-rhizobium symbiosis]. AB - A literature review synthesizes the data on physiological mechanisms of the influence of high doses of mineral nitrogen (nitrates and ammonium) on the formation and functioning of legume-rhizobium symbiosis. The participation of phytohormonal and phenolic metabolism and active forms of oxygen and nitrogen in the symbiosis is highlighted. Close connection between these metabolic processes in the formation and functioning of legume-rhizobium symbiosis under a redundant supply of plants by mineral nitrogen is underlined. PMID- 19548617 TI - [General structure of the mushroom body calyx in brachycera orthorrhapha flies (Diptera)]. AB - The mushroom body calyx in Brachycera Orthorrhapha flies is extremely diverse in the degree of development. In general, the calyx has the anterior, posterior, and dorsal lobes, as well as "sleeves" of glomerular neuropil surrounding Kenyon cell fibers. The anterior lobe of the calyx is found in all species studied. The most complex structure of the calyx is characteristic of the families Empididae and especially Bombyliidae, in which it has all the parts listed above. Brachycera Orthorrhapha flies have three fiber bundles of Kenyon cells, in contrast to four bundles in Drosophila flies. It is assumed that each mushroom body in Brachycera Orthorrhapha flies is formed by descendants of three single neuroblasts. PMID- 19548618 TI - [The butterflies (Lepidoptera, Diurna) of the boreal forests in southeastern Russia. II. Light coniferous forests]. AB - Species groupings formed in dark coniferous forests of southeastern Russia are analyzed by the example of butterflies. The species and chorological composition of these groupings are discussed. Classification analysis of species checklists for twelve local faunas is given. It is indicated that the applicability of a single faunistic category to all the dark coniferous forests of the studied region is doubtful. PMID- 19548619 TI - [Plasma membrane depolarization and activation of receptors for endogenous vasoconstrictors as possible mechanisms of potentiation of vasoconstrictive response to serotonin in traumatic shock in rats]. AB - The goal of this work was to study possible mechanisms underlying the potentiation of vasopressor response to serotonin observed in traumatic shock. Experiments with isolated aorta and mesenteric artery of the rat showed that vasoconstriction is caused by the activation of 5HT2A receptors. Agonists of 5HT1B, 5HT1D, 5HT2B, and 5HT4 receptors induced vasodilation. Agonists of 5HT1A receptors had a dual effect determined by interaction with alpha1-adrenergic receptors and 5HT1A receptors. Plasma membrane depolarization with 15 mM KCl increased the vasoconstrictive force in response to serotonin. This effect was determined by the ability of KCl to activate voltage-gated calcium channels, as a result of which the intracellular calcium stores are replenished. Inhibition of serotonin response by ketanserin, a 5HT2A receptor blocker, did not depend on the presence of 15 mM KCl. Constriction in response to serotonin was potentiated after its addition to vessels preconstricted with noradrenaline or endothelin-1. The constriction response partially retained in the presence of 2 x 10(-7) M ketanserin, which completely suppressed the serotonin-induced constriction of dilated vessels both at normal membrane potential and after plasma membrane depolarization. It can be assumed that noradrenalin and endothelin-1 alter the characteristics of 5HT2A receptors and possibly 5HT1A receptors as a result of their heterodimerization with the receptors for these vasoconstrictive hormones or interreceptor interaction at the level of signaling systems. Along with the potentiating effect of KCl, this mechanism may underlie the enhancement of vasopressor response to serotonin in shock. PMID- 19548620 TI - [Structure of component parasite communities in the grayling, Thymallus thymallus L. (Salmoniformes, Thymallidae), and minnow, Phoxinus phoxinus L. (Cypriniformes, Cyprinidae), from the upper reaches of the Pechora River]. AB - A comparative analysis of parasite communities in the grayling and minnow has been performed. These communities differ in the number of constituent species and in that the community of the minnow is dominated by one allogenic species, whereas that of the grayling usually has two dominants, both being autogenic species. The values of species diversity indices characterizing these communities are also different. Differences between the parasite communities of these fish species reflect the position of their hosts in the hydrobiocenosis. Species prevailing in both communities are classified as specialists, which is characteristic of parasite communities of the boreal-piedmont faunal complex. Both these communities consist of three species groups distinguished by their proportions in the total biomass, which indicates that parasitic communities are structured in a certain way and that the distribution of species in them in not random. PMID- 19548621 TI - [Effect of growth factors on morphogenesis of human keratinocytes in vitro]. AB - The effect of some growth factors on morphogenetic processes in the primary culture of human epidermal keratinocytes was studied in the model of formation of tubule-like structures in collagen gel. Culturing of keratinocytes in the presence of IGF and bFGF did not induce growth of tubule-like structures, whereas EGF, KGF, and HGF promoted the growth of three-dimensional epidermal outgrowths whose shape and size varied depending on the growth factor used. PMID- 19548622 TI - [Characterization of mycocin secreted by Rhodotorula colostri (Castelli) lodder]. AB - A Rhodotorula colostri strain has been isolated that shows fungicidal activity against related species of the genera Rhodosporidium, Sporidiobolus, and their anamorphs phylogenetically belonging to the order Sporidiobolales (Microbotryomycetes, Pucciniomycotina). The agent secreted has a molecular weight of about 4-5 kDa, is active at acid pH ranging from 3.5 to 5.5, and is thermolabile and protease-sensitive. PMID- 19548623 TI - [Microsporogenesis and pollen formation in Scotch pine (Pinus sylvestris L.)]. AB - The prolongation of a warm season during autumn, typical for the Siberia in recent 5-6 years, caused changes in the terms of reproductive processes in microsporangia of Scotch pine in the Krasnoyarsk region. The meiosis started in the autumn of the year, preceding the pollination, and comes to the end in next spring, whereas usually this process occurs in the spring of the year of pollination. In such conditions the division of microsporocytes accompanied with different abnormalities, causing various anomalies and the incomplete development of pollen grains. PMID- 19548624 TI - Seasonal levels of the major American cockroach allergen per a 9 (arginine kinase) in Bangkok and their relevance for disease severity. AB - Monitoring the levels of cockroach (CR) allergen in the environment has medical relevance as a clear dose response relationship between CR allergen exposure, sensitization and hospitalization has been reported. In this study, a cross sectional survey of the levels of a major American cockroach (Periplaneta americana) allergen, i.e. Per a 9 (arginine kinase) in dust samples collected in various seasons throughout the year 2007 from 76 houses of CR allergic Thai patients in the Bangkok metropolitan area were determined. A monoclonal antibody polyclonal antibody (MAb-PAb) based-sandwich ELISA was used. The MAb was specific to Per a 9 and the PAb was raised in a rabbit against the crude extract of P. americana. The detection limit of the assay was 122 pg of the allergen or 0.024 microg per gram of fine dust powder. The concentrations of Per a 9 were found to be highest during the winter months and lowest in summer. The levels of this CR allergen had a direct correlation with disease exacerbation; i.e. the majority of the CR allergic patients had their most severe clinical manifestations during winter. Moreover, the CR allergen levels were found to be higher in wood based houses than in concrete houses. PMID- 19548625 TI - Allergenicity of native/recombinant tropomyosin, per a 7, of American cockroach (CR), Periplaneta americana, among CR allergic Thais. AB - In this study, native tropomyosin (Per a 7) of American cockroach (CR), Periplaneta americana, caught in Thailand was purified. Also, gene sequence encoding full length tropomyosin of the CR was PCR amplified by using degenerate primers designed from gene sequences coding for P. americana tropomyosin of the database (Per a 7.0101 and Per a 7.0102; accession no.Y14854 and AF106961, respectively). Amino acid sequence deduced from the nucleotide sequence encoding P. americana tropomyosin of this study (GenBank accession no. FJ976895) had 98.59% identity with the sequences of Per a 7.0101 and Per a 7.0102 and was 97.18% identical to the Bla g 7 sequence of German cockroach, Blatella germanica (accession no. AF260897). The native and recombinant tropomyosins (approximately 34 kDa) were used as antigens in sandwich ELISA for detecting specific IgE in serum samples of 14 consented allergic patients who were positive by skin test to crude CR extract in comparison to 5 individuals who were skin test negative. It was found that 8 (57%) and 6 (43%) of the CR allergic patients gave positive IgE binding results to the native and the recombinant proteins, respectively, while none of the non-allergic counterparts was positive. Results of immunoblotting conformed to the ELISA results. Tropomyosin extracted from the P. americana caught in Thailand has potential as standard P. americana allergen in clinical monitoring of the allergic Thai patients. PMID- 19548626 TI - An analysis of skin prick test reactions in allergic rhinitis patients in Istanbul, Turkey. AB - This retrospective, population-based study reviewed skin prick test (SPT) results against various allergens of 1,552 patients with allergic rhinitis (AR) in the district of Kartal, Istanbul, Turkey. The skin prick tests yielded a positive result in 946 patients (60.9%). Seven hundred and forty-give (48%) patients had perennial AR, 558 (36%) perennial AR with seasonal exacerbations and 249 (16%) had seasonal AR. The prevalence of AR was highest in the age of 21-40 years with 48.7%. The allergen group with the highest SPT positivity was pollens at 44.3%, followed by molds at 38.4%, mites at 35.1%, and epithelia-insect at 30.8%. The strongest single allergen was Altemaria altemata with 33.3%, followed by Dermatophagoides farinae with 29.3%, Dermatophagoides pteronyssinus with 25.3% and a mix of four cereals (barley, maize, oat, wheat) with 25.2%. This information will help etiological research as well as the development of more efficient treatment plans for patients with allergic rhinitis in our country. PMID- 19548627 TI - Sleep disturbances and asthma control: a real life study. AB - The objective of the present study was to investigate the level of asthma control in real life and the relationship between Asthma Control Test (ACT) scores and sleep disturbances. Patients on control visits for bronchial asthma were asked to complete the ACT questionnaire and to answer 5 questions about their sleep quality (S5). It was found that asthma control was unsatisfactory in 44%. In all ACT classes the level of asthma control appeared to be inversely related to the presence of sleep disturbances: patients with good control reported less frequent and less severe sleep disturbances than uncontrolled subjects. However, a significant percentage of subjects (11-20%) with total control of asthma still had sleep disturbances that resulted in an impaired quality of life. Therefore in any asthma case with sleep disturbances a broader investigation is required whether that impairment is actually a result of asthma or some other co morbidity. Thus it would be useful if patients who reported sleep disturbances despite good/total control of asthma and rhinitis were managed with a holistic clinical approach and underwent nocturnal polysomnographic monitoring. PMID- 19548628 TI - Antisense interleukin-5 reduces eosinophil infiltration and hyperresponsiveness in an allergic asthma model. AB - Interleukin-5 (IL-5) involves in the development of airway inflammation and hyperresponsiveness through activation of eosinophils. Thus, inhibition of IL-5 expression seems to be an attractive approach for asthma therapy. In this study, an antisense IL-5 gene transferred by recombinant adeno-associated virus (asIL-5) was constructed to transfect murine allergic asthma model. Our results showed that asIL-5 efficiently inhibited the IL-5 mRNA expression and significantly attenuated the inflammation in lung tissues. Significant decreasing of eosinophils and inflammatory cells were found in peripheral blood and bronchoalveolar lavage fluid (BALF). In addition, significant inhibition of airway hyperresponsiveness (AHR) was also found in the mice treated with asIL-5. These observations demonstrate that antisense oligonucleotid against IL-5 delivered by adeno-associated virus system is possibly an efficacious therapeutic strategy for allergic asthma and other eosinophil-related disorders. PMID- 19548629 TI - Clinical and laboratory evaluation of periodically monitored Turkish children with IgG subclass deficiencies. AB - IgG subclass deficiencies are common immune system disorders during childhood. The aim of this retrospective study was to review clinical findings and laboratory results of patients with IgG subclass deficiencies in order to determine the changes in serum IgG subclass levels during follow-up, the percentage and time span until normalization of the IgG subclass levels to age corresponding normal levels, the type of infections incurred and the benefits of prophylaxis. Among the 59 pediatric patients reviewed, the most frequent defect was an IgG3 subclass deficiency (77%). Nine percent of the patients had an isolated IgG2 deficiency and 14% had an IgG2+G3 deficiency. The most common clinical presentations were recurrent upper respiratory tract infections, followed by pneumonia, acute gastroenteritis and urinary tract infections. Atopy was present in 15% of the patients. Ninety percent of the patients were given a prophylactic treatment (benzathine penicillin, oral antibiotics, oral bacterial lysate or intravenous immunoglobulin). The frequency of recurrent infections decreased from 13.4 +/- 7.4 per year to 5.7 +/- 3.9 in patients receiving a prophylactic regimen. Serum IgG subclass levels reached normal ranges in 30% of the patients in the IgG3 deficiency group and in 35.7% of the patients in the IgG2+G3 deficiency group. Patients with an isolated IgG2 deficiency did not reach age-related normal levels during the study period. Our study shows that IgG subclass levels may normalize in 30 to 40% of patients at about 6 years of age. We emphasize the need of monitoring IgG levels together with the clinical symptomatology in affected individuals and initiate preventive measures when appropriate. PMID- 19548630 TI - Atorvastatin attenuates TLR4-mediated NF-kappaB activation in a MyD88-dependent pathway. AB - Antigen presenting cells such as dendritic cells and macrophages have recently been detected in atherosclerotic plaques. Toll-like receptors expressed on the surface of these cells, have been implicated in ongoing inflammatory responses in the plaques. In this study, we investigated the anti-inflammatory effect of atorvastatin, a lipid lowering drug, via Toll-like receptor 4 (TLR4) in vitro, employing murine pro-B cell lines transfected with hTLR4/MD2 and MyD88/hTLR4/MD2 systems. The results showed that atorvastatin at 10 microM significantly attenuated NF-kappaB activation within 24 hours while at lower doses of 0.1 and 1 microM, treatment time had to be prolonged up to 48 hours for a significant inhibition to occur. The inhibition of NF-kappaB was also observed in a cell line cotransfected with MyD88 and TLR4 suggesting that the attenuation of NF-kappaB by atorvastatin occurred in a MyD88 dependent fashion. PMID- 19548631 TI - Differential gene expression profiles of lung epithelial cells exposed to Burkholderia pseudomallei and Burkholderia thailandensis during the initial phase of infection. AB - Burkholderia pseudomallei is the causative agent of melioidosis, and its infection usually affects patients' lungs. The organism is a facultative intracellular Gram-negative bacillus commonly found in soil and water in endemic tropical regions. Another closely related Burkholderia species found in soil and water is B. thailandensis. This bacterium is a non-pathogenic environmental saprophyte. B. pseudomallei is considerably more efficient than B. thailandensis in host cell invasion and adherence. A previous study by our group demonstrated that after successfully invading cells, there was no difference in the ability to survive and to replicate between both Burkholderia species in cultured A549 human lung epithelial cells. In this study, Human Affymetrix GeneChips were used to identify the difference in gene expression profiles of A549 cells after a 2-h exposure to B. pseudomallei and B. thailandensis. A total of 280 of 22,283 genes were expressed at higher levels in the B. pseudomallei-infected cells than in the B. thailandensis-infected cells, while 280 genes were expressed at lower levels in the B. pseudomallei-infected cells. Approximately 9% of these genes were involved in immune response and apoptosis. Those genes were further selected for gene expression analysis using reverse transcription PCR and/or real-time RT-PCR. The results of RT-PCR and real-time RT-PCR are in accordance with data from the microarray data in that bcl2 gene expression in the B. pseudomallei-infected cells was 2-fold higher than the level in the B. thailandensis-infected cells even though no apoptosis was seen in the infected cells. The levels of E selectin, ICAM-1, IL-11, IRF-1, IL-6, IL-1beta and LIF genes expression in the B. pseudomallei-infected cells were 1.5-5 times lower than in the B. thailandensis infected cells. However, both species stimulated the same level of IL-8 production from the tested epithelial cell line, and no difference in the ratio of adherent polymorphonuclear cells (PMNs) to infected A549 cells of both species was observed. Taken together, our results suggest that B. pseudomallei manipulates host response in favor of its survival in the host cell, which may explain the more virulent characteristics of B. pseudomallei when compared with B. thailandensis. PMID- 19548632 TI - Allergic rhinitis and co-morbid asthma: perspective from India -- ARIA Asia Pacific Workshop report. AB - In India, allergic rhinitis (AR) is considered to be a trivial disease, despite the fact that symptoms of rhinitis were present in 75% of children and 80% of asthmatic adults. Traditionally, AR was also divided into seasonal or perennial, based on the time of occurrence of symptoms during the year. The ARIA workshop report proposed that patients be categorized as "intermittent" and "persistent" while severity was classified as "mild" and "moderate-severe". Patients with AR, depending on their predominant symptom, can also be categorized as "sneezers runners" and "blockers". On sketching their clinical profile, it was observed that "blockers" had significantly higher sinusitis and had higher sensitization to fungi. Skin allergy testing in Indian adults showed that in patients with AR house dust mite (Dermatophagoides farinae) was the most common allergen. Studies conducted in India have shown that AR often restricts the patient's quality of life (QOL). It can affect the physical, psychological and social aspects of the patients' life and can also impact their functions at work. Furthermore, AR adversely affects sleep related QOL. Topical corticosteroids are now considered as the cornerstone of the treatment for AR. In spite of causing a major impact on the QOL in Indian patients, AR is rarely given the importance it deserves. PMID- 19548634 TI - Successful reduction of healthcare associated MRSA infection rates. PMID- 19548633 TI - Epidemiology and current status of allergic rhinitis and asthma in Thailand -- ARIA Asia-Pacific Workshop report. AB - The allergic diseases of the airway, i.e. allergic rhinitis and asthma, are on the increase in Thailand and their prevalence shows no signs of abating. When compared with a previous study, the incidence of wheezing had increased 4 fold (from 4.2% to 18.3%), and allergic rhinitis increased nearly 3 fold (from 17.9% to 44.2%). The results of the ISAAC phase III study revealed that the frequency of allergic diseases of the respiratory tract increased significantly from the ISAAC phase I survey performed in 1995; i.e. asthma increased from 12.2% to 14.5%, and allergic rhinitis from 37.9% to 50.6%. Allergic rhinitis exerts a major impact on the quality of life of Thai patients. The results of skin prick testing have indicated the leading causes of indoor (house-dust mites, house dust, cockroaches, dogs and cats) and outdoor pollen (Bermuda grass, para grass, sedge, careless weed) allergens. Molds (represented by Cladosporium), although prominent in an aeroallergen survey, returned a low percentage of positive skin prick reactions, and therefore, were considered low in allergenicity. In Thailand, there are clinical practice guidelines for both allergic rhinitis and asthma which are comparable to the international guidelines like ARIA and GINA. Sufficient kinds of pharmacotherapy are on the National List of Essential Drugs. Yet due to the limited number of trained allergists, many patients are seen by general physicians, and often, the appropriate diagnostic tests and treatments are not provided. In addition, the financial burden for quality health care may be prohibitive for those without private health insurance in spite of the implementation of a universal health care system for all Thai citizens, which is less than optimal. PMID- 19548635 TI - Microwave-mediated nonaqueous synthesis of quantum dots at moderate temperature. AB - The use of microwave irradiation to accelerate both inorganic and organic chemical reactions has attracted widespread attention. Generally, microwave mediated synthesis of quantum dots (QDs) has been conducted in aqueous solution. Here, using commercial diesel and glycerol as reaction medium, a microwave mediated nonaqueous method toward CdSe QDs with size-tunable photoluminescent properties produces oleic-acid-protected QDs at moderate reaction temperatures of 50-140 degrees C, which are much lower than the current temperature necessary for the synthesis of CdSe QDs in organic solvents. The appropriate condition optimization for high-quality CdSe QDs shows that different sizes of CdSe QDs with emission wavelengths between 450 and 600 nm have been synthesized through varying time, temperature, feed ratio, and reaction medium. PMID- 19548637 TI - Catalytic properties of cobalt(III)-oxo cubanes in the TBHP oxidation of benzylic alcohols. AB - Two series of oxo-bridged Co(III) complexes of the type Co(4)O(4)(O(2)CC(6)H(4) X)(4)(py)(4) (1) and Co(4)O(4)(O(2)CC(6)H(4)-X)(4)(4-Mepy)(4) (2), where X = H (a), Me (b), MeO (c), Cl (d), NO(2) (e), have been synthesized and characterized in detail. The molecular structures of the complexes consist of a cubelike Co(4)(mu(3)-O)(4) core having Co and O atoms at alternate vertices with carboxylato ligands bridging the Co(3+) ions along four face diagonals of the approximate cube. Nitrogen atoms from pyridyl ligands complete the distorted octahedral coordination around each Co(III). These neutral Co(III) complexes undergo a nearly reversible one-electron oxidation involving the redox couple [Co(4)(III)(mu(3)-O)(4)](4+) <--> [Co(3)(III)Co(IV)(mu(3)-O)(4)](5+) at potentials (approximately 0.7-1.0 V) that linearly depend on the electronic influence of X. The cobalt(III) clusters of types 1 and 2 have been found to effectively promote the TBHP oxidation of benzylic alcohols under homogeneous conditions to produce the corresponding carbonyl compounds. PMID- 19548636 TI - Rapid and efficient hydrophilicity tuning of p53/mdm2 antagonists. AB - The protein-protein interaction of p53 and mdm2 is an important anticancer target. The interface, however, is very hydrophobic and naturally results in very hydrophobic antagonists. We used the Orru three component reaction (O-3CR) along with a rapid and efficient, recently discovered amidation reaction to dramatically improve the water solubility of our recently discovered low molecular weight p53/mdm2 antagonists. Arrays of amides were synthesized with improved hydrophilicity and retainment and/or improvement of p53/mdm2 inhibitory activity. PMID- 19548638 TI - Efficient catalytic phosphate ester cleavage by binuclear zinc(II) pyrazolate complexes as functional models of metallophosphatases. AB - A series of dizinc(II) complexes based on the pyrazolate ligands 3-[(1E)-N hydroxyethanimidoyl]-4-methyl-1H-pyrazole-5-carboxylic acid (H(3)L(1)), (1E,1'E) 1,1'-(4-methyl-1H-pyrazole-3,5-diyl)diethanone dihydrazone (HL(2)), (E,E)-(4 methyl-1H-pyrazole-3,5-diyl)bis(methylmethanone) dioxime (H(3)L(3)), (E,E)-(4 phenyl-1H-pyrazole-3,5-diyl)bis(phenylmethanone) dioxime (H(3)L(4)), and 1H pyrazole-3,5-dicarboxylic acid (H(3)L(5)) have been synthesized and investigated as functional models of phosphoesterases, focusing on correlations between the hydrolytic activity and molecular parameters of the bimetallic core. Speciation of the various dizinc complexes in solution has been determined potentiometrically, and the structures in the solid state have been established by X-ray crystallography. The hydrolysis of two phosphoesters, an RNA model 2 hydroxypropyl-p-nitrophenyl phosphate (HPNP) and the pesticide paraoxon-ethyl (POE), promoted by the dinuclear phosphoesterase model complexes has been investigated in DMSO/buffered water (1:1) at 50 degrees C as a function of complex concentration, substrate concentration, and pH. Drastic differences in the hydrolytic activities of [Zn(2)(HL(1))(2)](0), [Zn(2)(L(2))(2)](2+), [Zn(2)(H(2)L(3))(2)](2+), and [Zn(2)(HL(5))(2)](2-) are observed and can be attributed to molecular peculiarities. Pyrazolate-bridged dinuclear zinc(II) complexes seem to provide a sufficient number of coordination sites for both activating the substrate and generating the nucleophile, where the phosphate esters are preferentially bound in a bidentate bridging fashion (in the case of HPNP) and in a monodentate fashion (in the case of POE). PMID- 19548639 TI - Organometallic complexes for nonlinear optics. 42. Syntheses, linear, and nonlinear optical properties of ligated metal-functionalized oligo(p phenyleneethynylene)s. AB - A combination of UV-vis-NIR spectroscopy, femtosecond Z-scan measurements, and time-dependent density functional theory (TD-DFT) calculations have been used to comprehensively investigate the linear optical and nonlinear optical (NLO) properties of pi-delocalizable metal-functionalized oligo(phenyleneethynylene)s. A range of unsymmetrically or symmetrically end-functionalized mono-, di-, tri-, penta-, hepta-, and nona(phenyleneethynylene)s were synthesized, with larger examples bearing varying numbers of 2,5-di(hexyloxy)phenyl groups to ensure sufficient solubility of the metal complex derivatives. The effect of incorporating varying numbers of solubilizing substituents in the OPE bridge, peripheral group modification, OPE lengthening, coligand variation, and metal location in the OPE on the linear optical properties has been established, with the first three molecular modifications resulting in significant changes in the optical absorption maxima. TD-DFT calculations reveal that the most intense transition in the linear optical spectra is localized on the OPE bridge and involves excitation from acetylenic to cumulenic molecular orbitals that are not greatly spatially separated from one another. The nonlinear optical properties are dominated by two-photon absorption, which for all but 1,4-{trans [RuCl(dppm)(2)]C[triple bond]C}(2)C(6)H(4) appears as a band around 11,400 cm(-1) and a sharp increase of nonlinear absorption at frequencies >17,000 cm(-1). Surprisingly, there is relatively little influence of the length of the OPE bridge on the magnitude of the two-photon absorption cross sections, which are in the range 300-1000 GM. PMID- 19548640 TI - Itinerant flat-band magnetism in hydrogenated carbon nanotubes. AB - We investigate the electronic and magnetic properties of hydrogenated carbon nanotubes using ab initio spin-polarized calculations within both the local density approximation (LDA) and the generalized gradient approximation (GGA). We find that the combination of charge transfer and carbon network distortion makes the spin-polarized flat-band appear in the tube's energy gap. Various spin dependent ground state properties are predicted with the changes of the radii, the chiralities of the tubes, and the concentration of hydrogen. It is found that strain or external electric field can effectively modulate the flat-band spin splitting and even induce an insulator-metal transition. PMID- 19548641 TI - A family of aligned C-curved nanoarches. AB - One-dimensional (1-D) nanomaterials are basic building blocks for the construction of nanoscale devices. However, the fabrication and alignment of 1-D nanomaterials with specific geometry and composition on a given substrate is a significant challenge. Herein we show a successful example of fabricating a family of aligned 1-D C-curved nanoarches of different compositions on an extended Si surface by a simple and scalable method. The nanoarches are made up of either single-crystalline Sn nanorods encapsulated in carbon nanotubes (CNTs), SnO(2) nanotubes, or CNTs. The aligned 1-D C-curved nanoarches of single crystalline Sn nanorods in CNTs are prepared first by a facile in situ reduction of SnO(2) nanoparticles under standard chemical vapor deposition conditions. Nanoarches of CNTs and SnO(2) nanotubes were then derived from the Sn@CNT nanoarches by acid etching and by calcination in air, respectively. PMID- 19548642 TI - Wavelength Sensitivity of Single Nanowire Excitation Polarization Anisotropies Explained through a Generalized Treatment of Their Linear Absorption. AB - We investigate the excitation polarization anisotropy of individual semiconductor nanowires (NWs) by monitoring their band edge emission above 680 nm in order to clarify the origin of their strong polarization response. Samples studied include both CdSe and CdSe/CdS core/shell nanowires grown using solution chemistry as well as analogous wires made via chemical-vapor-deposition (CVD). In the limit of optically thick wires, with radii above ~25 nm, we find NW optical responses consistent with the interaction between strong dielectric contrast influences and the onset of bulk-like behavior. Namely, a sizable wavelength dependence of the excitation polarization anisotropy (rho(exc)) exists when NW diameters become comparable to the wavelength of light inside the wire. As a consequence, pronounced rho(exc) rolloffs occur at short wavelengths. By contrast, thinner wires do not exhibit such wavelength dependencies, in agreement with earlier studies. We quantitatively explain observed wavelength sensitivities by modeling the NW as an absorbing dielectric cylinder under plane wave excitation. A comparison of predicted rho(exc)-values to experimental numbers shows good agreement and confirms the existence of wavelength-dependent rho(exc)-values in optically thick wires. Additional results of the model include generalized expressions for NW linear absorption cross-sections under parallel, perpendicular, and circularly polarized excitation. This study therefore adds to a growing body of knowledge about NW polarization anisotropies, specifically, their response in a size regime where dielectric contrast effects compete with the onset of bulk-like behavior. PMID- 19548643 TI - Nonadiabatic Ab initio multichannel quantum defect theory applied to absolute experimental absorption intensities in H(2). AB - The positions and intensities of the Q(N) (N = 1-4) X1 Sigma(g)+ --> nppi1 Pi(u)- (n = 2 to approximately 30) absorption transitions of H2 have been calculated by multichannel quantum defect theory. The computations are based on the quantum chemical ab initio clamped nuclei potential curves and absorption dipole transition moments for n = 2-4 of Wolniewicz and Staszewska (J. Mol. Spectrosc. 2003, 220, 45). The resulting Einstein spontaneous Einstein A coefficients are in good agreement with those derived from the absolute intensity measurements of Glass-Maujean et al. (Mol. Phys. 2007, 105, 1535). The results reveal widespread vibronic intensity perturbations in the Q(N) Rydberg series, whereas the line frequencies are comparatively little affected by nonadiabatic effects. PMID- 19548644 TI - Germylene energetics: electron affinities and singlet-triplet gaps of GeX(2) and GeXY species (X, Y = H, CH(3), SiH(3), GeH(3), F, Cl, Br, I). AB - A systematic investigation of the GeX(2) and GeXY species was carried out using the popular DFT functionals BLYP, B3LYP, and BHHLYP. Predicted are the singlet triplet energy gaps and four types of neutral-anion separations: adiabatic electron affinity (EA(ad)), zero-point vibrational energy corrected EA(ad(ZPVE)), vertical electron affinity (EA(vert)), and vertical detachment energy. The basis sets used for all atoms in this work are of double-zeta plus polarization quality with additional s- and p-type diffuse functions denoted DZP++, except for iodine where the 6-311G(d,p) basis set is used. The geometries are fully optimized with each functional independently. Vibrational frequency analyses were performed to compute zero-point energy corrections and to determine the nature of the stationary points. The geometries and the relative energies are discussed and compared with the carbon and silicon analogues. The EA(ad(ZPVE)) values (eV) obtained with the B3LYP functional range from 0.62 eV [Ge(CH(3))(2)] to 2.08 eV [Ge(GeH(3))(2)]. These results compare satisfactorily with the few available experiments, but most are reported for the first time. Similarly, the predicted singlet-triplet energy separations range from 13.8 kcal mol(-1) [Ge(SiH(3))(2)] to 85.0 kcal mol(-1) [GeF(2)]. Invariably, as one progresses down the periodic table C --> Si --> Ge, the "great divide" occurs between carbon and silicon. PMID- 19548645 TI - Structural and spectroscopic studies of the photophysical properties of benzophenone derivatives. AB - The effect a solvent has on the photophysical properties of a series of benzophenone derivatives, all FDA approved for use in sunscreens, is examined. Experimentally significant differences in the solvatochromic behavior are found to be dependent upon the substituents on the parent benzophenone molecule. The spectral trends do not appear to originate from only changes in the solvent polarity but indicate that specific solvent-solute interactions influence the absorbance energies of some benzophenones. Computational investigations examine the structure and electronic excitation energies of the molecules. Specific interactions of the solvent and solute are modeled to evaluate structural changes that result from solvent-solute complexation and the impact of the changes upon absorbance properties. The viability of an intramolecular excited state proton transfer is theoretically evaluated. The combination of experimental and computational analysis provides a more complete understanding of the molecular level origin of the unique photophysical properties of this class of UV absorbers. PMID- 19548646 TI - QTAIM analysis of ligand properties and mechanisms of tuning of 6-membered ring N heterocyclic carbenes in transition metal complexes through ring-substituent variation. AB - Tuning of the ligand properties of a series of N-heterocyclic carbenes has previously been achieved through the variation of substituents at the ring boron sites. Analysis of the topology of the electron density using the quantum theory of atoms in molecules (QTAIM) reveals sensitive changes in the integrated atomic properties and the curvature of the valence shell charge concentration of the carbene carbon (C2). Amino substitution induces greater ligand-to-metal sigma donation in the transition metal complex, which in turn weakens the attractive potential energy density on the interatomic surface of the trans-carbonyl group leading to the decrease in the trans C-O stretching frequency observed experimentally and theoretically. The distribution of charge concentrations within the inner-valence shell of cobalt in the transition metal complexes recovers the QTAIM analogs of ligand-to-metal sigma-donation and metal-to-ligand pi-back-donation. Investigation of the virial of the Ehrenfest force acting on the interatomic surfaces shows the operative mechanism of ligand tuning to be the inductive withdrawal of charge from the ring by nitrogen and subsequent back polarization of remaining charge toward C2. This mechanism is at odds with the orbital viewpoint of exo nitrogen-to-ring-boron pi-back-donation; however, it is the electronic forces that govern the bonding and charge distribution within a molecule in a stationary state. PMID- 19548647 TI - Coacervation of cationic gemini surfactant with weakly charged anionic polyacrylamide. AB - Coacervation of cationic gemini surfactant hexamethylene-1,6 bis(dodecyldimethylammonium bromide) (C(12)C(6)C(12)Br(2)) with 10% hydrolyzed polyacrylamide (PAM) has been observed and investigated by turbidity titration, isothermal titration calorimetry, dynamic light scattering and microscopy. Without any assistant additive, the coacervation takes place at very low surfactant concentration, and exists in a broad surfactant concentration range. The morphology of the coacervate sponge phase varies in pore size as a function of C(12)C(6)C(12)Br(2) concentration. The polymer/surfactant aggregates grow from soluble complexes with sizes smaller than 20 nm to micrometer during coacervation, and break up into soluble complexes of about 40 nm after coacervate redissolution. PMID- 19548648 TI - Truncated human betaB1-crystallin shows altered structural properties and interaction with human betaA3-crystallin. AB - The purpose of the study was to determine the effects of truncation of various regions of betaB1-crystallin on its structural properties and stability of heterooligomers formed by wild-type (WT) betaB1 or its deletion mutants with WT betaA3-crystallin. For these analyses, seven deletion mutants of betaB1 crystallin were generated with the following sequential deletions of either N terminal arm [betaB1(59-252)], N-terminal arm + motif I [betaB1(99-252)], N terminal arm + motif I + motif II [betaB1(144-252)], N-terminal arm + motif I + motif II + connecting peptide [betaB1(149-252)], C-terminal extension [betaB1(1 234)], C-terminal extension plus motif IV [betaB1(1-190)], or C-terminal extension + motif III + motif IV [betaB1(1-148)]. The betaB1-crystallin became water insoluble on the deletion of C-terminal extension and subsequent deletions of the C-terminal domain (C-terminal extension plus motifs III and IV) while it remained partially soluble on the deletion of the N-terminal domain (N-terminal arm plus motifs I and II). However, circular dichroism spectral analysis showed that the deletion of the N-terminal domain but not the C-terminal domain exhibited relatively greater structural changes in the crystallin. The deletion of the C-terminal domain resulted in a greater exposure and disturbance in the microenvironment of Trp-100, Trp-123, and Trp-126 (localized in the motif II), suggesting a relatively greater role of the C-terminal domain than the N-terminal domain in the structural stability of the crystallin. The deletion of the N terminal extension in betaB1 resulted in maximum exposure of hydrophobic patches and compact structure and in a maximum loss of subunit exchange with WT betaA3 crystallin compared to deletion of either the C-terminal extension, the N terminal domain, or the C-terminal domain. The thermal stability results of the heterooligomer of betaB1- plus betaA3-crystallins suggested that oligomers lose their stability on deletion of the C-terminal domain. Together, the results suggested that the N-terminal arm of betaB1-crystallin plays a major role in interaction with betaA3-crystallin during heterooligomer formation, and the solubility of betaB1-crystallin per se and that of the heterooligomer with betaA3 crystallin are dependent on the intact C-terminal domain of betaB1-crystallin. PMID- 19548650 TI - Solvatochromism in binary mixtures: first report on a solvation free energy relationship between solvent exchange equilibrium constants and the properties of the medium. AB - We have employed UV-vis spectroscopy in order to investigate details of the solvation of six solvatochromicindicators, hereafter designated as "probes", namely, 2,6-diphenyl-4-(2,4,6-triphenylpyridinium-1-yl) phenolate(RB); 4-[(E)-2 (1-methylpyridinium-4-yl)ethenyl] phenolate, MePM; 1-methylquinolinium-8-olate, QB;2-bromo-4-[(E)-2-(1-methylpyridinium-4-yl)ethenyl] phenolate, MePMBr, 2,6 dichloro-4-(2,4,6-triphenylpyridinium-1-yl) phenolate (WB); and 2,6-dibromo-4 [(E)-2-(1-methylpyridinium-4-yl)ethenyl] phenolate,MePMBr2, respectively. These can be divided into three pairs, each includes two probes of similar p kappa(a) in water and different lipophilicity. Solvation has been studied in binary mixtures, BMs, of water, W, with 12protic organic solvents, S, including mono- and bifunctional alcohols (2-alkoxyethanoles, unsaturated and chlorinated alcohols). Each medium was treated as a mixture of S, W, and a complex solvent, S W, formed by hydrogen bonding. Values of lambda max (of the probe intramolecular charge transfer) were converted into empirical polarity scales, ET(probe) in kcal/mol, whose values were correlated with the effective mole fraction of waterin the medium, chi W(effective). This correlation furnished three equilibrium constants for the exchange of solvents int he probe solvation shell; phi W/S (W substitutes S); phi S-W/W (S-W substitutes W), and phi S-W/S (S-W substitutes S), respectively. The values of these constants depend on the physicochemical properties of the probe and the medium. We tested, for the first time, the applicability of a new solvation free energy relationship: phi =constant + a alpha(BM) + b beta(BM) + s(pi* (BM) + d delta) + p log P (BM), where a, b, s, and p are regression coefficients; RBM,alpha (BM), beta(BM) and pi (BM) are solvatochromic parameters of the BM, delta is a correction term for pi*, and log P is an empirical scale of lipophilicity. Correlations were carried out with two-, three-, and four-medium descriptors.In all cases, three descriptors gave satisfactory correlations; use of four parameters gave only a marginal increase of the goodness of fit. For pi W/S, the most important descriptor was found to be the lipophilicity of the medium; for phi S-W/W and phi S-W/S, solvent basicity is either statistically relevant or is the most important descriptor. These responses are different from those of ET(probe) of many solvatochromic indicators in pure solvents, where the importance of solvent basicity is usually marginal, and can be neglected. PMID- 19548649 TI - Fluorescence correlation spectroscopy of phosphatidylinositol-specific phospholipase C monitors the interplay of substrate and activator lipid binding. AB - Phosphatidylinositol-specific phospholipase C (PI-PLC) enzymes simultaneously interact with the substrate, PI, and with nonsubstrate lipids such as phosphatidylcholine (PC). For Bacillus thuringiensis PI-PLC these interactions are synergistic with maximal catalytic activity observed at low to moderate mole fractions of PC (X(PC)) and maximal binding occurring at low mole fractions of anionic lipids. It has been proposed that residues in alpha-helix B help to modulate membrane binding and that dimerization on the membrane surface both increases affinity for PC and activates PI-PLC, yielding the observed PI/PC synergy. Vesicle binding and activity measurements using a variety of PI-PLC mutants support many aspects of this model and reveal that while single mutations can disrupt anionic lipid binding and the anionic lipid/PC synergy, the residues important for PC binding are less localized. Interestingly, at high X(PC) mutations can both decrease membrane affinity and increase activity, supporting a model where reductions in wild-type activity at X(PC) > 0.6 result from both dilution of the substrate and tight membrane binding of PI-PLC, limiting enzyme hopping or scooting to the next substrate molecule. These results provide a direct analysis of vesicle binding and catalytic activity and shed light on how occupation of the activator site enhances enzymatic activity. PMID- 19548652 TI - Influence of charged nanoparticles on colloidal forces: a molecular simulation study. AB - We employ the grand canonical Monte Carlo simulation technique to investigate the influence of charged nanoparticles (macro-ions) on the force between colloidal objects. Specifically, the structure and osmotic pressure of a system of screened Coulomb (Yukawa) particles confined between charged planar walls are simulated. We observe osmotic pressure to oscillate with wall separation and these oscillations to correspond to changes in the number of nanoparticle layers present in the slit pore. Using the Derjaguin approximation, we estimate the overall force between a colloidal sphere and a flat surface and compare our predictions to recent atomic force microscopy (AFM) results (Tulpar, A.; Van Tassel, P. R.; Walz, J. Y. Langmuir 2006, 22, 2876-2883). In excellent agreement with experiment, we find the wavelength of the force versus distance oscillations to scale as c(nu), with c being the bulk nanoparticle concentration and nu = 0.31 +/- 0.01; that is, slightly lower in magnitude from the expected value -1/3 based on average molecule spacing. By considering an order parameter measuring the extent to which neighboring particles form hexagonal symmetry, we show structural order within confined nanoparticle systems to be significantly enhanced as compared to that of bulk systems, despite being quite insensitive to wall separation. Wavelength scaling and order parameter analysis together suggest the confined macro-ion systems to be somewhat glasslike. PMID- 19548651 TI - Salt effects on surface-tethered peptides in solution. AB - The capability to manipulate proteins/peptide fragments at liquid-solid interfaces has led to tremendous applications in detectors and biotechnology. Therefore, understanding the detailed molecular behavior of proteins and peptides tethered on a hard material surface is an interesting and important topic. The inhomogeneity presented by surfaces as well as ions in the solution plays an important role in the thermodynamics and kinetics of the tethered proteins. In this study, we perform a series of molecular dynamics simulations of a pentapeptide RHSVV, a p53 epitope, tethered on a prepared microarray surface in various salt concentrations (0, 0.14, 0.5, and 1 M NaCl), as well as free in ionic solution (0, 0.5, and 1 M). The conformational space the tethered peptide visits largely overlaps with the free peptide in solution. However, surface tethering as well as the salt concentration changes both the thermodynamics and kinetics of the peptide. Frequent conformational changes are observed during the simulations and tend to be slowed down by both increasing the salt concentration and surface tethering. The local composition of ions at different salt concentrations is also compared between the tethered and free peptide. PMID- 19548653 TI - Protein segments with conformationally restricted amino acids can control supramolecular organization at the nanoscale. PMID- 19548654 TI - Proton affinities of some amino acid side chains in a restricted environment. AB - We investigate the dependence of proton affinity values of the side chains of amino acids such as Asp, Glu, His, Ser, and Thr on confinement in a single-walled carbon nanotube. The proton affinity values, estimated using the density functional theories (PW91/dnp and BLYP/dnp), are found to be highly sensitive toward confinement. We find that for both Asp and Glu, the proton affinity, while suspended inside the carbon nanotube, becomes much less in comparison to their respective gas phase values. In the case of His, Ser, and Thr side chains, on the other hand, the proton affinity inside the carbon nanotube becomes negative. Hydrogen bonding with neighboring polar groups is found to result in a marked increase in proton affinity inside the tube in all of the cases reported in this article. The increase is most remarkable in the case of His, Ser, and Thr side chains where the presence of polar neighboring groups within a hydrogen-bonding distance is found to augment the proton affinity value by more than 100 kcal mol( 1). PMID- 19548655 TI - Heterocyclic photochemistry in contrast with carbon behavior. Regioselective photochemical rearrangement of an azacyclohexadienone: mechanistic and exploratory organic photochemistry. AB - The Type-A photochemistry of cyclohexadienones is well-studied and follows a well established mechanistic pathway. One early example is the rearrangement of santonin to lumisantonin. Another example is the rearrangement of 4,4 diphenylcyclohexa-1,5-dienone. Remarkably, replacement of one carbon by nitrogen alters the reaction course to give a regioselective phenyl migration. PMID- 19548656 TI - Electrochemical synthesis of zinc nanoparticles via a metal-ligand-coordinated vesicle phase. AB - Two salt-free Zn(2+)-ligand-coordinated vesicle phases were prepared from the mixtures of alkyldimethylamine oxide (C(n)DMAO, n = 14 and 16, i.e., C(14)DMAO and C(16)DMAO) and zinc laurate [(CH(3)(CH(2))(10)COO)(2)Zn] in aqueous solution. The two salt-free Zn(2+)-ligand-coordinated vesicle phases were subsequently used as the templating media for fabricating zinc nanoparticles on an indium-doped tin oxide (ITO) electrode via electrodeposition. Influence of temperature, direct current density, and composition of the surfactant mixtures on the average diameter and size distribution of Zn nanoparticles was investigated in more detail. The structure of Zn@ITO was determined by scanning electron microscope and energy-dispersive X-ray spectroscopy analysis. Monodispersed Zn nanoparticles on ITO electrode (ZnNPs@ITO) with different average diameters were obtained, and the distribution can be controlled. Finally, ZnO nanostructures on the ITO substrate were prepared from the as-synthesized Zn@ITO nanoparticles through electrochemical surface oxidation. The preparation of nanostructured Zn and ZnO particles by our established method may pave the way for a new templating route from metal-ligand-coordinated vesicles. PMID- 19548657 TI - Mechanism of S(H)2 reactions of disulfides: frontside vs backside, stepwise vs concerted. AB - Density functional theory calculations indicate that the S(H)2 reactions of disulfides with alkyl or aryl radicals take place via concerted backside displacement. The activation energies for reactions of Me* with RSSR (R = Me, Et, (i)Pr, (t)Bu) increase with the size of R, since larger R groups prevent the formation of an ideal geometry for SOMO-LUMO overlap. Frontside transition states can also be located, but these lie at least 11 kcal mol(-1) above the corresponding backside transition states. PMID- 19548658 TI - Impact of transporters in oral absorption: a case study of in vitro and in vivo organic anion absorption. AB - A key determinant for oral bioavailability of a drug candidate is the intestinal epithelial permeation of the drug candidate. This intestinal permeation may be affected by interactions on membrane transporters expressed in the intestinal epithelial cells. The purpose of the present study was to investigate whether transporters were involved in the intestinal absorption of an organic anion A275 and to compare the impact of interactions related to transporters in the Caco-2 cell model versus the in vivo rat model of intestinal absorption. In both models, it was investigated whether intestinal permeation of A275 was concentration dependent and affected by inhibitors or competitive organic anions. Interactions related to transporters in intestinal permeation was clearly demonstrated in the Caco-2 cell model but was not directly evident for in vivo rat absorption. However, an observed biphasic in vivo absorption and a large intervariability between rats might mask a dose-dependent absorption of A275. To avoid these suggested interactions, a dose of at least 10 mg/kg, which saturates the intestinal transporters involved in A275 absorption, should be administered, but at doses below that the risk of such drug interactions should be taken into account. PMID- 19548659 TI - Binding of Cu(II) to human alpha-synucleins: comparison of wild type and the point mutations associated with the familial Parkinson's disease. AB - The Cu(II)-alpha-synuclein interaction has been invoked as an important process in the pathogenesis of Parkinson's disease. Herein, we report binding constants and stoichiometry under near-physiological conditions for the binding of Cu(II) to human alpha-synuclein. Specifically, we compare the binding of Cu(II) to wild type (WT) protein and two separate single mutation proteins that are associated with familial Parkinson's diseases: A30P and A53T. Cu(II) binds to all three alpha-synuclein proteins with a 1:1 stoichiometry. The Cu(II) binding constants, however, vary among the proteins studied. Cu(II) binding to WT and A53T at 37 degrees C is similar with a pH-dependent binding constant (K) of approximately 2.4 x 10(9) and approximately 4.8 x 10(9) M(-1) at pH 7.2 and 7.4, respectively. Cu(II) binding to A30P, however, exhibits two binding constants. The major binding site of A30P, characteristic of >90% of the bound Cu(II), has binding constants of 1.6 x 10(9) and 3.6 x 10(9) M(-1) at pH 7.2 and 7.4, respectively, slightly lower ( approximately 70%) than that characteristic of WT or A53T at the corresponding pH. The second less populated binding exhibited by A30P has a large binding constant, approximately 10(10) M(-1). Our size exclusion analysis ruled out the contribution of protofibrils to the strong Cu(II) binding. Previous studies indicated that A30P had a larger proportion of intermediate species (e.g., small oligomeric species, such as dimers and trimers) relative to WT and A53T. Thus, we propose that the high affinity site is attributed to the binding of Cu(II) to those small oligomeric species. PMID- 19548660 TI - Biological and biomedical applications of two-dimensional vibrational spectroscopy: proteomics, imaging, and structural analysis. AB - In the last 10 years, several forms of two-dimensional infrared (2DIR) spectroscopy have been developed, such as IR pump-probe spectroscopy and photon echo techniques. In this Account, we describe a doubly vibrationally enhanced four-wave mixing method, in which a third-order nonlinear signal is generated from the interaction of two independently tunable IR beams and an electron polarizing visible beam at 790 nm. When the IR beams are independently in resonance with coupled vibrational transitions, the signal is enhanced and cross peaks appear in the spectrum. This method is known as either DOVE (doubly vibrationally enhanced) four-wave mixing or EVV (electron-vibration-vibration) 2DIR spectroscopy. We begin by discussing the basis and properties of EVV 2DIR. We then discuss several biological and potential biomedical applications. These include protein identification and quantification, as well as the potential of this label-free spectroscopy for protein and peptide structural analysis. In proteomics, we also show how post-translational modifications in peptides (tyrosine phosphorylation) can be detected by EVV 2DIR spectroscopy. The feasibility of EVV 2DIR spectroscopy for tissue imaging is also evaluated. Preliminary results were obtained on a mouse kidney histological section that was stained with hematoxylin (a small organic molecule). We obtained images by setting the IR frequencies to a specific cross-peak (the strongest for hematoxylin was obtained from its analysis in isolation; a general CH(3) cross peak for proteins was also used) and then spatially mapping as a function of the beam position relative to the sample. Protein and hematoxylin distribution in the tissue were measured and show differential contrast, which can be entirely explained by the different tissue structures and their functions. The possibility of triply resonant EVV 2DIR spectroscopy was investigated on the retinal chromophore at the centre of the photosynthetic protein bacteriorhodopsin (bR). By putting the visible third beam in resonance with an electronic transition, we were able to enhance the signal and increase the sensitivity of the method by several orders of magnitude. This increase in sensitivity is of great importance for biological applications, in which the number of proteins, metabolites, or drug molecules to be detected is low (typically pico- to femtomoles). Finally, we present theoretical investigations for using EVV 2DIR spectroscopy as a structural analysis tool for inter- and intramolecular interaction geometries. PMID- 19548661 TI - Temperature-dependent mechanisms for the dynamics of protein-hydration waters: a molecular dynamics simulation study. AB - Molecular dynamics simulations are performed to study the temperature-dependent dynamics and structures of the hydration shells of elastin-like and collagen-like peptides. For both model peptides, it is consistently observed that, upon cooling, the mechanisms for water dynamics continuously change from small-step diffusive motion to large-step jump motion, the temperature dependence of water dynamics shows a weak crossover from fragile behavior to strong behavior, and the order of the hydrogen-bond network increases. The temperature of the weak crossover from fragile to strong behavior is found to coincide with the temperature at which maximum possible order of the hydrogen-bond network is reached so that the structure becomes temperature independent. In the strong regime, the temperature dependence of water translation and rotational dynamics is characterized by an activation energy of E(a) approximately 0.43 eV, consistent with results from previous dielectric spectroscopy (DS) and nuclear magnetic resonance (NMR) studies on protein hydration waters. At these temperatures, a distorted pi-flip motion about the 2-fold molecular symmetry axes, i.e., a water-specific beta process, is an important aspect of water dynamics, at least at the water-peptide interfaces. In addition, it is shown that the hydration waters exhibit pronounced dynamical heterogeneities, which can be traced back to a strong slowdown of water motion in the immediate vicinity of peptide molecules due to formation of water-peptide hydrogen bonds. PMID- 19548662 TI - The structure of the protonated adenine dimer by infrared multiple photon dissociation spectroscopy and electronic structure calculations. AB - The infrared multiple photon dissociation (IRMPD) spectrum of electrosprayed adenine proton-bound dimers were recorded in the gas-phase inside the cell of a Fourier transform ion cyclotron resonance spectrometer coupled to a tunable optical parametric oscillator/amplifier infrared laser. While gas-phase B3LYP/6 31+G(d,p) calculations indicate that the four lowest isomers are essentially isoenergetic, comparisons of the experimental and predicted IR spectra suggest that only two of the four isomers are observed in the experiment. However, computed solvation effects, as modeled using both a polarizable continuum model and microsolvation with five explicit water molecules, preferentially stabilize these two observed isomers, consistent with the interpretation of the IRMPD spectra. This work shows that for these small species the solvent-phase structure is preserved. It also demonstrates the potential danger of using gas-phase calculations to predict the structures of gaseous ions born in solution, such as those from an electrospray source. PMID- 19548663 TI - Iterative accumulation multiplexing Fourier transform ion cyclotron resonance mass spectrometry. AB - A multiplexed tandem mass spectrometry (MS/MS) technique known as iterative accumulation multiplexing (IAM) has been implemented on a hybrid quadrupole Fourier transform ion cyclotron resonance mass spectrometer (Q-FTICR-MS). The IAM experiment resulted in obtaining MS/MS spectra for six analytes in two MS/MS experiments while characteristic resolving power and mass measurement accuracies were maintained. Parent-product ion correlations were graphically represented in a "ratiogram" where each product ion is encoded with a ratio unique to the parent ion from which it was formed. This is the first example of multiplexed MS on a FTICR instrument where the ions are encoded externally to the ICR cell. By performing the encoding external to the ICR cell, one set of ions can be encoded while the previous set of ions is being analyzed in the cell, maximizing the use of the continuous ion current emanating from the electrospray ionization source. PMID- 19548664 TI - Theoretical insights into the interaction mechanism between proteins and SWCNTs: adsorptions of tripeptides GXG on SWCNTs. AB - Adsorptions of nine tripeptides GXG, ranging from negatively (D) and positively (K) charged, to hydrophilic (N and S), and to hydrophobic (G, V, F, W, and Y) residues, on the two cluster models (C(54)H(18) and C(54)) of (10,0) single walled carbon nanotubes (SWCNTs) are systemically investigated with the MPWB1K and MP2 methods. The solvent effects are taken into account with the implicit CPCM model. The objective is to provide novel insights into the interaction mechanism between proteins and SWCNTs. Results reveal that the adsorption strength of two charged tripeptides is greatly affected by the solvent effect and the hydrogen saturability of the SWCNT models. In the gas phase, on the surface of C(54)H(18), GKG has the strongest adsorption (adsorption energy (AE): -29.3 kcal/mol at the MP2 level), whereas the adsorption of the negatively charged GDG is the strongest on C(54) (AE: -30.4 kcal/mol with MP2). However, because of strong solvation, the adsorptions of the charged residues (D and K) on both C(54)H(18) and C(54) surfaces in aqueous solution are either rather weak or even unbound. The two neutral hydrophilic residues (N and S) exhibit adsorptions on C(54)H(18) in the gas phase (AE: -3.3 and -4.2 kcal/mol), yet are unable to adsorb on SWCNTs in aqueous solution (AE: +0.3 kcal/mol at MP2+CPCM). The five hydrophobic residues present relatively strong adsorption on SWCNTs, especially for the three aromatic residues (GFG, GYG, and GWG), regardless of the CNT model and whether they are in the gas phase or solution. These results indicate that in general the aromatic groups of proteins would play a very important role on functionalizing CNTs, which basically supports the relevant experimental observations. In addition, the electron correlation is essential for adsorptions of GXG on pristine SWCNTs, and the three aromatic residues have the highest electron correlation effects. The present investigation provides strong evidence that for the functionalization of CNTs via proteins it is most likely that hydrophobic interaction and van der Waals are the dominant driving forces. PMID- 19548665 TI - Accelerator mass spectrometry targets of submilligram carbonaceous samples using the high-throughput Zn reduction method. AB - The high-throughput Zn reduction method was developed and optimized for various biological/biomedical accelerator mass spectrometry (AMS) applications of mg of C size samples. However, high levels of background carbon from the high-throughput Zn reduction method were not suitable for sub-mg of C size samples in environmental, geochronology, and biological/biomedical AMS applications. This study investigated the effect of background carbon mass (mc) and background 14C level (Fc) from the high-throughput Zn reduction method. Background mc was 0.011 mg of C and background Fc was 1.5445. Background subtraction, two-component mixing, and expanded formulas were used for background correction. All three formulas accurately corrected for backgrounds to 0.025 mg of C in the aerosol standard (NIST SRM 1648a). Only the background subtraction and the two-component mixing formulas accurately corrected for backgrounds to 0.1 mg of C in the IAEA C6 and -C7 standards. After the background corrections, our high-throughput Zn reduction method was suitable for biological (diet)/biomedical (drug) and environmental (fine particulate matter) applications of sub-mg of C samples (> or = 0.1 mg of C) in keeping with a balance between throughput (270 samples/day/analyst) and sensitivity/accuracy/precision of AMS measurement. The development of a high-throughput method for examination of > or = 0.1 mg of C size samples opens up a range of applications for 14C AMS studies. While other methods do exist for > or = 0.1 mg of C size samples, the low throughput has made them cost prohibitive for many applications. PMID- 19548667 TI - Multibed needle trap devices for on site sampling and preconcentration of volatile breath biomarkers. AB - To facilitate their use in trace gas analysis, the adsorption capacity of needle trap devices (NTDs) was increased by combining three adsorbent materials and increasing total adsorbent amount. The use of 22 gauge needles, application of internally expanding desorptive flow technique without cryofocusation and a new on site alveolar sampling method for NTDs provided sensitivity in the parts per trillion range of VOC concentrations without loosing precision or linearity. LODs were 0.4 ng/L for isoprene, 0.5 ng/L for dimethyl sulphide, 0.9 ng/L for 2 butenal, 1.0 ng/L for hexane, 1.2 ng/L for pentane, 2.3 ng/L for hexanal, 5.3 ng/L for pentanal, and 8.3 ng/L for acetone. R of calibration curves were consistently >0.98. Loss of volatile aldehydes during storage for 7 days was less than 10%. Needle trap devices packed with more than one adsorbent material represent a promising alternative to SPE and SPME for analysis of volatile organic compounds in the low parts per billion/parts per trillion range. Crucial problems of clinical breath analysis concerning sensitivity of analytical methods, limited stability, and decomposition of breath compounds during sampling and storage could be solved. PMID- 19548666 TI - Wireless, remote-query, and high sensitivity Escherichia coli O157:H7 biosensor based on the recognition action of concanavalin A. AB - Escherichia coli O157:H7 is detected using a remote-query (wireless, passive) magnetoelastic sensor platform to which a 1 microm thick layer of Bayhydrol 110 and then a layer of functionalized mannose is applied. The multivalent binding of lectin concanavalin A (Con A) to the E. coli surface O-antigen and mannose favors the strong adhesion of E. coli to the mannose-modified magnetoelastic sensor; E. coli is rigidly and strongly attached on the mannose-modified sensor through Con A, which works as a bridge to bind E. coli to the mannose-modified sensor surface. As E. coli is bound to the sensor, its resonance frequency shifts, enabling quantification of E. coli concentration with a limit of detection of 60 cells/mL and a linear logarithmic response range of 6.0 x 10(1) to 6.1 x 10(9) cells/mL. The analysis can be directly conducted without incubation and completed in 3 h or less. PMID- 19548669 TI - Competing magnetism and superconductivity in Na(x)CoO2 at half doping. AB - We report the effect of topotactic replacement of Na(+) by H(3)O(+) in Na(0.5)CoO(2). Hydronium ions keep the Co oxidation state constant, increasing the interlayer distance. The result is a progressive destruction of the spin density wave characteristic of half-doped Na(x)CoO(2) and ultimately the induction of superconductivity. This implies that superconductivity occurs closer to Co(3+)/Co(4+) approximately = 1 than previously thought and that its development competes with the development of the magnetic phase. PMID- 19548668 TI - Identification, characterization, and bioconversion of a new intermediate in valanimycin biosynthesis. AB - The antibiotic valanimycin is a naturally occurring azoxy compound isolated from Streptomyces viridifaciens. Detailed investigations have shown that valanimycin is derived from L-valine and L-serine via the intermediacy of O-(L seryl)isobutylhydroxylamine. Sequence analysis of the valanimycin biosynthetic genes provides relatively few clues concerning the nature of the later stages of the pathway. Two exceptions are provided by the vlmJ and vlmK genes. The translation product of vlmJ exhibits similarity to diacylglycerol kinases, while the translation product of vlmK exhibits a weak similarity to the MmgE/PrpD superfamily of proteins. This superfamily includes 2-methylcitrate dehydratase. This communication describes the isolation and structure elucidation of valanimycin hydrate from vlmJ and vlmK mutants of S. viridifaciens. Additional studies have shown that the conversion of valanimycin hydrate into valanimycin by S. viridifaciens requires both the vlmJ and vlmK genes and that VlmJ catalyzes the ATP-dependent phosphorylation of the hydroxyl group of valanimycin hydrate prior to VlmK-catalyzed dehydration. PMID- 19548670 TI - Ion-exchangeable, electronically conducting layered perovskite oxyfluorides. AB - Cation-exchangeable d(0) layered perovskites are amenable to intercalation, exfoliation, and a variety of topochemical reactions, but they lack the interesting electronic and magnetic functionalities of mixed-valent perovskites. Conversely, electronically and magnetically interesting layered perovskites lack scope in terms of interlayer chemistry. To bridge this gap, the insulating, cation-exchangeable layered perovskites RbLaNb(2)O(7), KCa(2)Nb(3)O(10), and NaYTiO(4) were reacted with poly(tetrafluoroethylene) under inert atmosphere conditions to yield layer perovskites in which some of the oxygen is substituted by fluorine. In the fluorinated materials, the B-site cations are reduced to a mixed-valent state without introducing oxygen vacancies into the anion sublattice. The resulting electronically conducting solids can be exposed to air and water and even ion-exchanged in acid without oxidation of the B-site cations. Electronic transport measurements on the air-stable RbLaNb(2)O(6)F reveal room temperature conductivity (2-7 x 10(2) ohms x cm) via a variable-range hopping mechanism, which is not substantially changed after aqueous proton exchange to H(1-x)Rb(x)LaNb(2)O(6)F (x approximately = 0.2). PMID- 19548671 TI - Simultaneous degradation of organophosphates and 4-substituted phenols by Stenotrophomonas species LZ-1 with surface-displayed organophosphorus hydrolase. AB - Organophosphorous hydrolase (OPH) was expressed onto the surface of a Stenotrophomonas species (LZ-1), capable of simultaneously degrading 4 substituted phenols, using the N- and C-terminal domains of ice nucleation protein (INPNC) as an anchoring motif for the first time. The engineered strain LZ-1 could degrade p-nitrophenyl-substituted organophosphates as well as their hydrolytic product, PNP, rapidly. Especially, addition of 4-CP (below 0.8 mM) significantly accelerated the complete degradation of above organophosphates (47.1, 34.0, and 40% reduction of time of paraoxon, parathion, and methyl parathion, respectively) through the accelerated degradation of PNP due to enhanced cell growth supported by 4-CP as the carbon source. OPH could be surface displayed at a high level without inhibition of cell growth and OPH activity in the presence of 4-CP. In soil samples, strain LZ-1 could also remove these compounds successfully. Functional display of heterologous proteins on the surface of indigenous bacteria could provide a promising technology for effective bioremediation of sites contaminated with mixed organic pollutants. PMID- 19548673 TI - Controllable columnar organization of positively charged polycyclic aromatic hydrocarbons by choice of counterions. AB - A novel columnar organization of ionic complexes based on 9 phenylbenzo[1,2]quinolizino[3,4,5,6-fed]phenanthridinylium (PQP) has been achieved via ionic self-assembly. These complexes represent the first family of polycyclic aromatic hydrocarbons containing a large charged aromatic core with controllable columnar organization in both the crystalline and liquid-crystalline phases. The single-crystal structure of the ionic complex with a benzenesulfonate anion exhibits a staggered dimer arrangement of PQP cations that further establishes columnar superstructures. The use of sulfonate anions with long alkyl tails leads to well-ordered discotic columnar mesophases with an identical staggered packing of the PQP cations. PMID- 19548672 TI - Narrow-band absorption-enhanced quantum dot/J-aggregate conjugates. AB - We report narrow-band absorption enhancement of semiconductor nanocrystals via Forster resonance energy transfer from cyanine J-aggregates. These J-aggregated dyes associate electrostatically with short quantum-dot (QD) surface ligands in solution. Energy transfer efficiencies approach unity for this light sensitization and result in a 5-fold enhancement in the QD excitation near the J aggregate absorption maximum. Because a thin layer of J-aggregates attenuates the same amount of light (at peak absorbance) as a far thicker film of monomer dye, these absorption-enhanced materials may have applications in light-sensitizing applications such as photodetection and optical down-conversion. PMID- 19548674 TI - Effects of salinity changes on the growth of Dunaliella salina and its isozyme activities of glycerol-3-phosphate dehydrogenase. AB - Dunaliella salina could survive in media containing a wide range of NaCl concentrations ranging from about 0.05 M to saturation (around 5.5 M). Glycerol is an important osmolyte when Dunaliella survive in various salt environments, and G3pdh is a key enzyme in glycerol metabolism. The osmotic response of D. salina was investigated by studying its cell growth, glycerol content change, and isozyme activity of glycerol-3-phosphate dehydrogenase (G3pdh) in different salinities. Results showed that 2.0 M NaCl was the optimal salinity for the growth of D. salina, in which condition the highest glycerol content of 64.02 +/- 3.21 (mean +/- SD) microg/mL was detected. D. salina could rapidly increase or decrease glycerol contents to adapt to hypoosmotic or hyperosmotic environments. The glycerol content declined 52.05% when salinity was changed from 2.0 to 0.5 M NaCl, and the glycerol content increased 43.61% when salinity was increased from 2.0 to 5.0 M NaCl. In the isozyme electrophoresis assay two kinds of isozymes, G3pdh and superoxide dismutase (Sod), were detected synchronously. Interestingly, it was first found that there are five isozymes of G3pdh in D. salina. G3pdh-2 mainly takes effect in moderate to high salinities, whereas the other four isozymes take effect in low salinities, which may provide an important clue for future research on osmoregulation mechanisms. PMID- 19548677 TI - Synthesis of periodic mesoporous coesite. AB - Periodic mesoporous coesite was obtained by a modified nanocasting process from a periodic mesoporous silica SBA-16/C composite at a pressure of 12 GPa and 350 degrees C. PMID- 19548676 TI - Differential charge polarization of axial histidines in bacterial reaction centers balances the asymmetry of the special pair. AB - In photosynthesis, light energy is transformed into chemical energy that sustains most forms of life on earth. Solid-state NMR spectroscopy in conjunction with density functional theory modeling can resolve electronic structure down to the atomic level in large membrane proteins. In this work, we have used this technique to address the mechanisms underlying the photochemical reactivity of the special pair in the bacterial reaction center. For charge separation, the electrostatics is important, as the Coulomb barrier must be overcome. On the basis of (15)N NMR data, we resolve a subtle charge-balancing mechanism in the ground state by an axial histidine that is connected to the central Mg(2+) on one side and hydrogen-bonded on the other side. Formation of the hydrogen bond between BChl-a-His and H(2)O leads to a difference in electron density relative to the separate BChl-a-His and H(2)O fragments, with excess positive charge on the imidazole ring. This can lower the kinetic barrier for accommodating the different length scales of electron and proton transfer for separation of spin and charge in a bidirectional proton-coupled electron-transfer mechanism. PMID- 19548678 TI - Turning MIL-53(Al) redox-active by functionalization of the bridging OH-group with 1,1'-ferrocenediyl-dimethylsilane. AB - The postsynthetic functionalization of the bridging OH-group between two metal centers of the secondary building units of MIL-53(Al) with 1,1'-ferrocenediyl dimethylsilane is the proof of principle of a new, selective functionalization method of the "inorganic" part of the porous coordination polymer (PCP). The functionalized material was active in the liquid-phase oxidation of benzene to phenol as a test reaction for redox activity. PMID- 19548675 TI - Red wine protects against ethanol-induced oxidative stress in rat liver. AB - Ethanol consumption may be deleterious to the liver. However, alcoholic beverages contain, besides ethanol (EtOH), complex chemical mixtures that can modify EtOH's adverse effects. Red wine (RW) is rich in polyphenolic antioxidants, often reported as hepatoprotective agents. This study aimed to investigate the effects of 6 months of RW ingestion on hepatic oxidative stress and inflammation. Six month-old Wistar rats were treated with RW or EtOH; controls were pair-fed. EtOH increased 8-hydroxy-2'-deoxyguanosine and decreased reduced and oxidized glutathione. These animals also displayed stimulated superoxide dismutase, catalase, and glutathione reductase activities. RW treatment decreased malondialdehyde and reduced glutathione levels. Glutathione-S-transferase and selenium-dependent glutathione peroxidase activities were stimulated and glutathione reductase activity was inhibited by RW intake. No modifications were detected in nuclear factor-kappa B or alkaline phosphatase activities. EtOH consumption induced fibrosis in portal spaces and hepatocyte lipid accumulation that were absent with RW treatment. This paper highlights the importance of RW nonalcoholic components and the relevance of biological matrix in the study of EtOH oxidative effects. PMID- 19548679 TI - A novel immunomodulatory protein from Poria cocos induces Toll-like receptor 4 dependent activation within mouse peritoneal macrophages. AB - Poria cocos is an important Oriental medical fungus with multiple functionalities, yet its bioactive substances and the mechanisms involved have not been fully characterized. A novel immunomodulatory protein (P. cocos immunomodulatory protein; PCP) was purified from the dried sclerotium of P. cocos (Schw.) Wolf using DE-52 cellulose and gel filtration chromatography. Chromatography and electrophoresis results indicated that the native PCP (35.6 kDa) is a disulfide-linked heterodimeric glycoprotein consisting of 14.3 and 21.3 kDa subunits with N- and O-glycosylation. PCP was capable of stimulating RAW 264.7 macrophages in vitro through the induction of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta) as well as the regulation of nuclear factor-kappa B (NF-kappaB)-related gene expression. In primary mouse macrophages, PCP directly activated peritoneal cavity macrophages to induce Toll like receptor 4 (TLR4)-mediated myeloid differentiation factor 88 (MyD88) dependent signaling. This study demonstrated the cell surface interactions of PCP with TLR4 and the capacity of PCP for TLR4 tyrosine phosphorylation. Results obtained with peritoneal macrophages from TLR4-deficient C57BL/10ScN mice revealed that PCP-induced activation and PCP cell surface binding were significantly attenuated. Moreover, enzymatic deglycosylation decreased PCP mediated responses, indicating that the glycosylated portion of PCP was a key factor in PCP signaling through TLR4 in peritoneal macrophages. These findings suggest that PCP is a new potential immune stimulator within P. cocos and that TLR4 is primarily responsible for PCP signaling in murine macrophages. PMID- 19548680 TI - Calcium leads to further increase in glycine-enhanced extracellular secretion of recombinant alpha-cyclodextrin glycosyltransferase in Escherichia coli. AB - Overexpression of recombinant genes in Escherichia coli and targeting recombinant proteins to the culture medium are highly desirable for the production of industrial enzymes. However, a major barrier is inadequate secretion of recombinant protein across the two membranes of E. coli cells. In the present study, we have attempted to circumvent this secretion problem of the recombinant alpha-cyclodextrin glycosyltransferase (alpha-CGTase) from Paenibacillus macerans strain JFB05-01. It was found that glycine, as a medium supplement, could enhance the extracellular secretion of recombinant alpha-CGTase in E. coli. In the culture with glycine at the optimal concentration of 150 mM, the alpha-CGTase activity in the culture medium reached 23.5 U/mL at 40 h of culture, which was 11 fold higher than that of the culture in regular TB medium. A 2.3-fold increase in the maximum extracellular productivity of recombinant alpha-CGTase was also observed. However, further analysis indicated that glycine supplementation exerted impaired cell growth as demonstrated by reduced cell number and viability, increased cell lysis, and damaged cell morphology, which prevented further improvement in overall enzyme productivity. Significantly, Ca(2+) could remedy cell growth inhibition induced by glycine, thereby leading to further increase in the glycine-enhanced extracellular secretion of recombinant alpha CGTase. In the culture with 150 mM glycine and 20 mM Ca(2+), both extracellular activity and maximum productivity of recombinant enzyme were 1.5-fold higher than those in the culture with glycine alone. To the best of our knowledge, this is the first article about the synergistic promoting effects of glycine and Ca(2+) on the extracellular secretion of a recombinant protein in E. coli. PMID- 19548681 TI - Application of natural antimicrobials for food preservation. AB - In this review, antimicrobials from a range of plant, animal, and microbial sources are reviewed along with their potential applications in food systems. Chemical and biochemical antimicrobial compounds derived from these natural sources and their activity against a range of pathogenic and spoilage microorganisms pertinent to food, together with their effects on food organoleptic properties, are outlined. Factors influencing the antimicrobial activity of such agents are discussed including extraction methods, molecular weight, and agent origin. These issues are considered in conjunction with the latest developments in the quantification of the minimum inhibitory (and noninhibitory) concentration of antimicrobials and/or their components. Natural antimicrobials can be used alone or in combination with other novel preservation technologies to facilitate the replacement of traditional approaches. Research priorities and future trends focusing on the impact of product formulation, intrinsic product parameters, and extrinsic storage parameters on the design of efficient food preservation systems are also presented. PMID- 19548682 TI - Microsphere-based rolling circle amplification microarray for the detection of DNA and proteins in a single assay. AB - We describe a high-density microarray for simultaneous detection of proteins and DNA in a single test. In this system, Rolling Circle Amplification (RCA) was used as a signal amplification method for both protein and nucleic acid detection. The microsphere sensors were tested with synthetic DNA and purified recombinant protein analytes. The target DNA sequence was designed from a highly conserved gene that encodes the outer membrane protein P6 (OMP-P6) of both typeable and nontypeable strains of Haemophilus influenzae. The proinflammatory mediators IL-6 and IL-8 were selected as target proteins. Capture antibodies were first immobilized on fluorescently encoded microspheres. The microspheres were then loaded into the etched microwells of an imaging optical fiber bundle. A sandwich assay was performed for target proteins IL-6 and IL-8 using biotin-labeled secondary antibodies. Biotinylated capture DNA probes were then attached to the detection antibodies via an avidin bridge. A padlock probe, complementary to the target sequence, was subsequently hybridized to the capture probe. In the presence of the target sequence, the padlock probe was ligated, and this circular sequence was used for RCA. Following RCA, multiple fluorescently labeled signal probes were hybridized to each amplified sequence, and the microarray was imaged using an epi-fluorescence microscope. With this assay, detection limits down to 10 fM and 1 pM were achieved for proteins and target DNA, respectively. In addition to this new approach for detecting both protein and DNA in a single test using RCA, the limit of detection for IL-8 and IL-6 was improved by 3 orders of magnitude compared to similar microsphere-based assays. PMID- 19548683 TI - Demetalation of silver(III) corrolates. AB - Several procedures for the demetalation of silver(III) corrolates have been tested. Acidic conditions induce removal of the silver ion but they can also promote concomitant oxidation of the corrole nucleus to an isocorrole species, the degree of which will depend upon the specific acidic media. This oxidation cannot be completely avoided by addition of hydrazine, particularly in the case of 3-NO(2) substituted complexes which are quantitatively converted into the corresponding 3-NO(2), 5-hydroxy isocorroles upon silver ion removal. Several beta-nitro isocorrole products were isolated, and one was structurally characterized. Electrochemical and chemical reductive methods for silver(III) corrolates demetalation were then tested with the aim to avoid the formation of isocorroles. While reaction with sodium borohydride was shown to be quite effective to demetalate unsubstituted silver corrolates this was not the case for the beta-nitro derivatives where the peripheral nitro group is reduced by borohydride giving the corresponding 3-amino free base corrole species. For the beta-nitro corrole silver complexes, a successful approach was obtained using DBU/THF solutions which afforded the 3-NO(2) corrole free-base compound as a single reaction product in good yield. These conditions were also effective for unsubstituted corroles although longer reaction times were necessary in this case. To study in greater detail the corrole demetalation behavior, selected Ag(III) derivatives were characterized by cyclic voltammetry in pyridine, and the demetalation products spectrally characterized after controlled potential reduction in a thin-layer spectroelectrochemical cell. PMID- 19548684 TI - Stability studies on astaxanthin extracted from fermented shrimp byproducts. AB - To the best of our knowledge, stability studies on astaxanthin contained in carotenoproteins extracted from lactic acid fermented shrimp byproduct have never been reported. Carotenoprotein powder, containing 1% free astaxanthin, was subjected to oxidation factors of illumination, oxygen availability, and temperature, using synthetic astaxanthin as a control. The individual effects as well as first and second degree interactions were studied on natural and synthetic free astaxanthin stability. Air and full light were the two individual factors with the highest effects on astaxanthin oxidation. Sixty-two and 46% natural and synthetic astaxanthin, respectively, oxidized when exposed to air for 8 weeks of storage, whereas 35 and 28% of natural and synthetic astaxanthin, respectively, oxidized under full light. Ninety-seven and 88% of natural and synthetic astaxanthin, respectively, oxidized under a combination of full light, air, and 45 degrees C at 8 weeks of storage. Storage in the dark, nonoxygen, and 25 degrees C were the treatments that efficiently minimized astaxanthin oxidation. Natural astaxanthin from fermented shrimp byproduct presented moderate stability levels. Although natural astaxanthin oxidized faster than the synthetic pigment, its stability may improve by antioxidant and polymer addition. PMID- 19548685 TI - The effect of bacterial strain and aging on the secondary volatile metabolites produced during malolactic fermentation of tannat red wine. AB - During malolactic fermentation (MLF), lactic acid bacteria influence aroma and flavor of wines by the production of volatile metabolites and the modification of aroma compounds derived from grapes and yeasts. In an effort to isolate these bacteria properties as advantages for winemaking, this study aimed to assess the relative contribution of two aspects: the effects of lactic acid bacteria activity on the volatiles compounds in Tannat wines and the consequences of aging in bottle on aroma compounds produced during MLF. To our knowledge, this is the first report related to the effect of wine aging in bottle on the aroma chemical compounds produced by MLF. Solid phase extraction complemented with chromatographic techniques was used to study the wine aroma compounds. A sensory evaluation of the wines was also performed through descriptive methods. We demonstrated modifications in the concentration of acetates, ethyl esters, and other secondary metabolites during MLF. Major sensorial differences between wines that had undergone MLF were also noted. In addition, some modifications detected in the composition of Tannat wines as a consequence of the aging in bottle contributed to the change in differences between wines with and without MLF and furthermore between strains. These changes probably influence its fruity character. PMID- 19548686 TI - Mono- and bis- methyltrioxorhenium(VII) complexes with salen ligands: synthesis, properties, applications. AB - Methyltrioxorhenium(VII) (MTO) forms 1:1 (mono-) or/and 2:1 (bis-) complexes with salen ligands, undergoing a hydrogen transfer from a ligand-bound OH-group to a ligand nitrogen atom. Some complexes show good stability both in the solid state and in solution, while others must be kept at low temperatures under an argon atmosphere. X-ray crystallography shows distorted trigonal bipyramidal structures of all examined complexes in the solid state, this structure being due to the steric demands of the ligands, with the methyl group of MTO residing in the apical sites in the cis position. Temperature-dependent proton NMR data indicate that the coordination between salen ligands and MTO at low temperatures is considerably stronger than at room temperature. Density functional theory calculations have been performed to find approximate structures for all described complexes and to try to find a rationale for the preferred formation of mono- versus bis-MTO complexes. The formation of mono- or bis-MTO adducts is dependent on both the steric and the electronic influence of the respective salen ligands. The catalytic performance is strongly influenced by the ring substitution. Two MTO molecules coordinated to one salen ligand lead to an additional boost of catalytic activity because there is not only double the amount of catalytic centers present but also a "ligand enhanced" activity increase. PMID- 19548687 TI - Synthesis, binding, and modeling studies of new cytisine derivatives, as ligands for neuronal nicotinic acetylcholine receptor subtypes. AB - The availability of drug affecting neuronal nicotinic acetylcholine receptors (nAChRs) may have important therapeutic potential for the treatment of several CNS pathologies. Pursuing our efforts on the systematic structural modification of cytisine and N-arylalkyl and N-aroylalkyl cytisines were synthesized and tested for the displacement of [(3)H]-epibatidine and [(125)I]-alpha-bungarotoxin from the most widespread brain nAChRs subtypes alpha(4)beta(2) and alpha(7), respectively. While the affinity for alpha(7) subtype was rather poor (K(i) from 0.4 to >50 microM), the affinity for alpha(4)beta(2) subtype was very interesting, with nanomolar K(i) values for the best compounds. The N-substituted cytisines were docked into the rat and human alpha(4)beta(2) nAChR models based on the extracellular domain of a molluscan acetylcholine binding protein. The docking results agreed with the binding data, allowing the detection of discrete amino acid residues of the alpha and beta subunits essential for the ligand binding on rat and human nAChRs, providing a novel structural framework for the development of new alpha(4)beta(2) selective ligands. PMID- 19548688 TI - Synthesis of ruthenium carbonyl complexes with phosphine or substituted Cp ligands, and their activity in the catalytic deoxygenation of 1,2-propanediol. AB - A ruthenium hydride with a bulky tetra-substituted Cp ligand, (Cp(i)(Pr(4)))Ru(CO)(2)H (Cp(i)(Pr(4)) = C(5)(i-C(3)H(7))(4)H) was prepared from the reaction of Ru(3)(CO)(12) with 1,2,3,4-tetraisopropylcyclopentadiene. The molecular structure of (Cp(i)(Pr(4)))Ru(CO)(2)H was determined by X-ray crystallography. The ruthenium hydride complex (C(5)Bz(5))Ru(CO)(2)H (Bz = CH(2)Ph) was similarly prepared. The Ru-Ru bonded dimer, [(1,2,3 trimethylindenyl)Ru(CO)(2)](2), was produced from the reaction of 1,2,3 trimethylindene with Ru(3)(CO)(12), and protonation of this dimer with HOTf gives {[(1,2,3-trimethylindenyl)Ru(CO)(2)](2)-(mu-H)}(+)OTf (-). A series of ruthenium hydride complexes CpRu(CO)(L)H [L = P(OPh)(3), PCy(3), PMe(3), P(p-C(6)H(4)F)(3)] were prepared by reaction of Cp(CO)(2)RuH with added L. Protonation of (Cp(i)(Pr(4)))Ru(CO)(2)H, Cp*Ru(CO)(2)H, or CpRu(CO)[P-(OPh)(3)]H by HOTf at -80 degrees C led to equilibria with the cationic dihydrogen complexes, but H(2) was released at higher temperatures. Protonation of CpRu[P(OPh)(3)](2)H with HOTf gave an observable dihydrogen complex, {CpRu[P-(OPh)(3)](2)(eta(2)-H(2))}(+)OTf ( ) that was converted at -20 degrees C to the dihydride complex {CpRu[P(OPh)(3)](2)(H)(2)}(+)OTf (-). These Ru complexes serve as catalyst precursors for the catalytic deoxygenation of 1,2-propanediol to give n-propanol. The catalytic reactions were carried out in sulfolane solvent with added HOTf under H(2) (750 psi) at 110 degrees C. PMID- 19548689 TI - Energy gaps in supramolecular functionalized graphene nanoribbons. AB - The electronic structure characteristics of supramolecular functionalization of graphene nanoribbons with pi-conjugated polymers are investigated using first principles density functional theory. Noncovalent polymer functionalization leads to distinct changes in the electronic properties, particularly the band gaps of metallic and semimetallic graphene nanoribbons. A detailed analysis of band alignments reveals a profound level hybridization for ribbons with various shaped edges and spin density waves near the edges of zigzag ribbons. The extracted planar polymer conformations and the disappearance of the metallic behavior are in conformity with experimental observations. PMID- 19548690 TI - Macrophyllin-type bicyclo[3.2.1]octanoid neolignans from the leaves of Pleurothyrium cinereum. AB - Four new macrophyllin-type bicyclo[3.2.1]octanoid neolignans, (7S,8R,3'S,5'R) Delta(8')-5,5',3'-trimethoxy-3,4-methylenedioxy-2',3',4',5'-tetrahydro-2',4' dioxo-7.3',8.5'-neolignan (cinerin A), 1, (7R,8R,3'S,4'R,5'R)-Delta(8')-4' hydroxy-5,5'-dimethoxy-3,4-methylenedioxy-2',3',4',5'-tetrahydro-2'-oxo-7.3',8.5' neolignan (cinerin B), 2, (7S,8R,3'R,4'S,5'R)-Delta(8')-4'-hydroxy-5,5',3' trimethoxy-3,4-methylenedioxy-2',3',4',5'-tetrahydro-2'-oxo-7.3',8.5'-neolignan (cinerin C), 3, and (7S,8R,2'R,3'S,5'R)-Delta(8')-2'-hydroxy-5,5'-dimethoxy-3,4 methylenedioxy-2',3',4',5'-tetrahydro-4'-oxo-7.3',8.5'-neolignan (cinerin D), 4, along with the known diterpene kaurenoic acid 5, were isolated from the leaves of Pleurothyrium cinereum. The structures and configuration of these compounds were determined by extensive spectroscopic analysis. Cinerins A-D (1-4) were tested for their inhibition efficacy of platelet activating factor (PAF)-induced aggregation of rabbit platelets. Compound 3 was the most potent PAF antagonist. Compounds 1-5 were tested against Mycobacterium tuberculosis (H(37)Rv strain) using the MABA method. Compound 5 induced 91.3% growth inhibition at 50 microg mL(-1). Compounds 1-5 showed no significant inhibitory activity against some Gram positive and Gram-negative bacteria by the agar-well diffusion method. PMID- 19548691 TI - Facile and scalable synthesis of the fused-ring heterocycles thieno[3,2 b]thiophene and thieno[3,2-b]furan. AB - An optimized synthetic methodology which allows for efficient and scalable access to the important fused-ring heterocycle thieno[3,2-b]thiophene and the first reported isolation of thieno[3,2-b]furan is presented. The properties of thieno[3,2-b]furan were assessed through a detailed analysis of the NMR data and an investigation of the chemical reactivity. Thieno[3,2-b]furan is chemically robust and offers good selectivity toward functionalization at the 2-position via bromination and the 5-position via deprotonation. PMID- 19548692 TI - Mononuclear platinum(II) complexes incorporating kappa2-carboxylate ligands: synthesis, structure, and reactivity. AB - The chloro-bridged dimer [(PC)Pt(mu-Cl)](2) (PC = [((t)Bu(2)P-o-C(6)H(4)) CMe(2)](-)) reacts with silver(I) acetate to yield the monomeric kappa(2)-acetate complex [(PC)Pt(kappa(2)-OOCMe)]. Its trifluoroacetate analogue exists in solution as a mixture of the monomer and two isomeric dimers, and crystallizes as the approximately C(2)-symmetrical dimer [(PC)Pt(mu-OOCCF(3))](2). NMR experiments and density functional theory calculations show that the monomeric species are favored by bulky ligands and electron-rich carboxylates. [(PC)Pt(OOCCF(3))] undergoes a ligand-dehydrogenation reaction in hot trifluoroacetic acid to yield [(PCC)Pt(OOCCF(3))(2)] (PCC = ((t)Bu(2)P-o C(6)H(4))-C(Me)=CH(2)). PMID- 19548694 TI - Optical response of liquid acetonitrile at ambient conditions: the dynamical dielectric behavior from ab initio calculations. AB - We probe the linear optical properties of the neat liquid acetonitrile (CH(3)CN) at ambient conditions using ab initio density functional theory. Uncorrelated structures extracted from Monte Carlo simulation are employed to efficiently calculate average electronic properties. It becomes evident that condensation leads to a conduction band with a large degree of dispersion, which is consistent with the description of dipolar liquids. This allows an interpretation of the dielectric spectrum based on the electronic structure of liquid CH(3)CN, and clearly shows the influence of intermolecular interactions in the absorption features. We find that the lowest-lying excitation of the condensed phase occurs at 7.8 eV, which is reasonable as compared to the 8-9.5 eV absorption region measured in the gas phase. PMID- 19548693 TI - Diterpenoids from the Mediterranean brown alga Dictyota sp. evaluated as antifouling substances against a marine bacterial biofilm. AB - Four new cyclized diterpenes, one xenicane (1) and three dolabellanes (2-4), were isolated, along with seven previously reported metabolites [3beta-hydroxydilophol (5), dictyols E (6) and C (7), hydroxycrenulide (8), 9-acetoxy-15-hydroxy-1,6 dollabelladiene (9), hydroxyacetyldictyolal (10), and fucoxanthin], from a Mediterranean species of Dictyota sp. collected in Le Brusc Lagoon (French Mediterranean coast). Their structures, as well as their relative configurations, were determined through extensive spectrometric (IR, HRESIMS, 1D and 2D NMR) data analysis and molecular modeling studies and by comparison with those reported in literature. Some of the isolated metabolites were evaluated for their antiadhesion activity against a marine bacterial biofilm (Pseudoalteromonas sp. D41). PMID- 19548695 TI - Stability of the Two Au-S Binding Modes in Au(25)(SG)(18) Nanoclusters Probed by NMR and Optical Spectroscopy. AB - This report presents a detailed investigation on the structural stability of the Au(25)(SG)(18) nanoclusters (where, -SG represents glutathionate) with a focus on the oxidation resistance and thermal stability of the Au-S bonds in the cluster. Two types of Au-S binding modes were previously indentified in the crystal structure of phenylethylthiolate-capped Au(25) clusters, and the Au(25)(SG)(18) nanoclusters have been confirmed to adopt the same structure as Au(25)(SCH(2)CH(2)Ph)(18) in our previous works. Herein, NMR in combination with optical spectroscopy revealed some distinct differences in both antioxidation and thermal stability of the two Au-S binding modes in the Au(25)(SG)(18) nanocluster, that is, the mode I thiolate ligands (total 12 ligands) exhibit a much higher stability than the mode II ligands (total 6 ligands). Upon the basis of this major observation as well as an abundant [Au(25)S(12)](-) species detected in laser desorption ionization (LDI) mass spectrometry analysis of Au(25)(SG)(18) clusters, a metastable species similar to Au(25)(SG)(12) (i.e., a loss of six type II ligands of -SG) may exist in the processes of oxidation and thermal treatment. Overall, this work provides a deep insight into the intriguing structure of thiolate-capped Au(25) clusters, which will benefit future studies pertaining to the potential applications of such nanoclusters. PMID- 19548696 TI - Organohalogen and organophosphorous pesticide method for ginseng root--a comparison of gas chromatography-single quadrupole mass spectrometry with high resolution time-of-flight mass spectrometry. AB - A method has been developed for the analysis of 170 organohalogen and organophosphorous pesticides, isomers, and metabolites in dried ground ginseng root. Pesticides were extracted with ethyl acetate and purified with gel permeation chromatography (GPC) and primary/secondary amine modified silica (PSA)/graphitized carbon black (GCB) combination SPE column. Each purified pesticide extract was determined by both gas chromatography single quadrupole mass spectrometry using selected ion monitoring (GC-qMS-SIM) and by gas chromatography high resolution time-of-flight mass spectrometry (GC-HR-TOFMS). The geometric mean LOQs using the qMS and TOFMS were 4 and 3 ng/g ginseng, respectively. Mean recoveries from ginseng were 83, 79, and 75% with standard deviations of 4, 5, and 3%, respectively, for 25, 100, and 500 ng/g using GC-qMS SIM. Mean recoveries using GC-HR-TOFMS were 93, 85, and 81% with mean standard deviations of 7, 7, and 8% for 25, 100, and 500 ng/g, respectively. Seven dried ginseng root products were found to contain combinations of the following pesticides: dacthal, diazinon, DDT, hexachlorobenzene, iprodione, lindane, procymidone, and quintozene (1-460 ng/g). No significant differences were found in the concentrations measured for these pesticides on commercial ginsengs using either of the two GC/MS techniques. PMID- 19548697 TI - Treatise on the measurement of molecular masses with ion mobility spectrometry. AB - The ability to separate isotopes by high-resolution ion mobility spectrometry techniques is considered as a direct means for determining mass at ambient pressures. Calculations of peak shapes from the transport equation show that it should be possible to separate isotopes for low-mass ions (<200) by utilizing heavy collision gases and high-resolution ion mobility analyzers. The mass accuracy associated with this isotopic separation approach based on ion mobility separation is considered. Finally, we predict several isotopes that should be separable. PMID- 19548699 TI - Electron-deficient triphenylene and trinaphthylene carboximides. AB - A series of triphenylene and trinaphthylene carboximides were synthesized mainly through Diels-Alder reactions between the in situ generated radialenes and N alkylmaleimides. High electron affinity was obtained due to the attachment of electron-withdrawing carboximide groups. These disklike molecules can assemble into one-dimensional columnar structures in solid state and their mesophase properties were characterized by DSC, POM, and X-ray diffraction. Long-range ordered columnar phase was observed, qualifying them as potential n-type semiconductors in electronic devices. PMID- 19548698 TI - C-H bond functionalization via hydride transfer: synthesis of dihydrobenzopyrans from ortho-vinylaryl akyl ethers. AB - The hydride transfer initiated cyclization ("HT-cyclization") of aryl alkyl ethers, which leads to direct coupling of sp(3) C-H bonds and activated alkenes, is reported. Readily available salicylaldehyde derived ethers are converted in one step to dihydrobenzopyrans, an important class of heteroarenes frequently found in biologically active compounds. This process has not been previously reported, in contrast to known HT-cyclizations of the corresponding tert-amines ("tert-amino effect" reactions). PMID- 19548700 TI - Approaching the Hartree-Fock limit by perturbative methods. AB - We describe perturbative methods for improving finite-basis Hartree-Fock calculations toward the complete-basis limit. The best method appears to offer quadratic error reduction and preliminary numerical applications demonstrate that remarkably accurate Hartree-Fock energies can be obtained. PMID- 19548701 TI - Controlling molecular rotational population by wave-packet interference. AB - We propose a control scheme for selecting populations of molecular rotational states by wave-packet interference. A series of coherent rotational wave packets is created by nonadiabatic rotational excitation of molecules using two strong femtosecond laser pulses. By adjusting the time delay between the two laser pulses, constructive or destructive interference among these wave packets enables the population to be enhanced or suppressed for a specific rotational state. The evolution of the rotational wave packet with selected populations produces interference patterns with controlled spatial symmetries. This method provides an approach to prepare a molecular ensemble with selected quantum-state distributions and controlled spatial distributions under field-free condition. PMID- 19548702 TI - Auger recombination and excited state relaxation dynamics in Hg(n)(-) (n=9-20) anion clusters. AB - Using femtosecond time-resolved photoelectron imaging, electron-hole pairs are created in size-selected Hg(n)(-) anion clusters (n=9-20), and the subsequent decay dynamics are measured. These clusters eject electrons via Auger decay on time scales of 100-600 fs. There is an abrupt increase in the Auger decay time for clusters larger than Hg(12)(-), coinciding with the onset of the transition from van der Waals to covalent bonding in mercury clusters. Our results also show evidence for subpicosecond excited state relaxation attributed to inelastic electron-electron and electron-hole scattering as well as hole-induced contraction of the cluster. PMID- 19548703 TI - Strong fields induce ultrafast rearrangement of H atoms in H2O. AB - H atoms in H(2)O are rearranged by strong optical fields generated by intense 9.3 fs laser pulses to form H(2)(+). This atomic rearrangement is ultrafast: It occurs within a single laser pulse. Quantum-chemical calculations reveal that H(2)(+) originates in the (1)A state of H(2)O(2+) when the O-H bond elongates to 1.15 a.u. and the H-O-H angle becomes 120 degrees. Bond formation on the ultrafast time scale of molecular vibrations (10 fs for H(2)(+)) and in strong fields has hitherto not been reported. PMID- 19548705 TI - Langevin thermostat for rigid body dynamics. AB - We present a new method for isothermal rigid body simulations using the quaternion representation and Langevin dynamics. It can be combined with the traditional Langevin or gradient (Brownian) dynamics for the translational degrees of freedom to correctly sample the canonical distribution in a simulation of rigid molecules. We propose simple, quasisymplectic second-order numerical integrators and test their performance on the TIP4P model of water. We also investigate the optimal choice of thermostat parameters. PMID- 19548704 TI - Formation of a dual hydrogen bond in the N-H...C=O moiety in the indole-(N methylacetamide)1 cluster revealed by IR-dip spectroscopy with natural bond orbital analysis. AB - IR-dip spectra in the NH stretch regions have been measured for the S(0) state of the indole/N-methylacetamide 1:1 clusters (Ind-NMA(1)). We identified two structural isomers of Ind-NMA(1) that possess an N-H...O=C hydrogen bond. The redshifts of the NH stretch fundamental of the indole moieties in Ind-NMA(1) are larger than that for Ind-(H(2)O)(1) [Carney, Hagemeister, and Zweir, J. Chem. Phys. 108, 3379 (1998)], indicating that the strength of the N-H...O=C hydrogen bond in Ind-NMA(1) is stronger than that of the N-H...O-H hydrogen bond in Ind (H(2)O)(1). On the basis of the natural bond orbital analysis we suggest that two lone pair orbitals of the O atoms in the NMA moiety form a dual hydrogen bond with the NH group designated by N-H:::O=C. Owing to the dual nature of the N H:::O=C hydrogen bond its strength in Ind-NMA(1) is larger than that of the N H...O-H hydrogen bond in Ind-(H(2)O)(1). PMID- 19548706 TI - The initial and final states of electron and energy transfer processes: diabatization as motivated by system-solvent interactions. AB - For a system which undergoes electron or energy transfer in a polar solvent, we define the diabatic states to be the initial and final states of the system, before and after the nonequilibrium transfer process. We consider two models for the system-solvent interactions: A solvent which is linearly polarized in space and a solvent which responds linearly to the system. From these models, we derive two new schemes for obtaining diabatic states from ab initio calculations of the isolated system in the absence of solvent. These algorithms resemble standard approaches for orbital localization, namely, the Boys and Edmiston-Ruedenberg (ER) formalisms. We show that Boys localization is appropriate for describing electron transfer [Subotnik et al., J. Chem. Phys. 129, 244101 (2008)] while ER describes both electron and energy transfer. Neither the Boys nor the ER methods require definitions of donor or acceptor fragments and both are computationally inexpensive. We investigate one chemical example, the case of oligomethylphenyl 3, and we provide attachment/detachment plots whereby the ER diabatic states are seen to have localized electron-hole pairs. PMID- 19548707 TI - A phase-field approach to no-slip boundary conditions in dissipative particle dynamics and other particle models for fluid flow in geometrically complex confined systems. AB - Dissipative particle dynamics (DPD) is an effective mesoscopic particle model with a lower computational cost than molecular dynamics because of the soft potentials that it employs. However, the soft potential is not strong enough to prevent the DPD particles that are used to represent the fluid from penetrating solid boundaries represented by stationary DPD particles. A phase-field variable, phi(x,t), is used to indicate the phase at point x and time t, with a smooth transition from -1 (phase 1) to +1 (phase 2) across the interface. We describe an efficient implementation of no-slip boundary conditions in DPD models that combines solid-liquid particle-particle interactions with reflection at a sharp boundary located with subgrid scale accuracy using the phase field. This approach can be used for arbitrarily complex flow geometries and other similar particle models (such as smoothed particle hydrodynamics), and the validity of the model is demonstrated by DPD simulations of flow in confined systems with various geometries. PMID- 19548708 TI - Computation of potential energy surfaces with the multireference correlation consistent composite approach. AB - A multireference composite method that is based on the correlation consistent Composite Approach (ccCA) is introduced. The developed approach, multireference ccCA, has been utilized to compute the potential energy surfaces (PESs) of N(2) and C(2), which provide rigorous tests for multireference composite methods due to the large multireference character that must be correctly described as the molecules dissociate. As well, PESs provide a stringent test of a composite method because all components of the method must work in harmony for an appropriate, smooth representation across the entire surface. PMID- 19548709 TI - Simultaneous escaping of explicit and hidden free energy barriers: application of the orthogonal space random walk strategy in generalized ensemble based conformational sampling. AB - To overcome the pseudoergodicity problem, conformational sampling can be accelerated via generalized ensemble methods, e.g., through the realization of random walks along prechosen collective variables, such as spatial order parameters, energy scaling parameters, or even system temperatures or pressures, etc. As usually observed, in generalized ensemble simulations, hidden barriers are likely to exist in the space perpendicular to the collective variable direction and these residual free energy barriers could greatly abolish the sampling efficiency. This sampling issue is particularly severe when the collective variable is defined in a low-dimension subset of the target system; then the "Hamiltonian lagging" problem, which reveals the fact that necessary structural relaxation falls behind the move of the collective variable, may be likely to occur. To overcome this problem in equilibrium conformational sampling, we adopted the orthogonal space random walk (OSRW) strategy, which was originally developed in the context of free energy simulation [L. Zheng, M. Chen, and W. Yang, Proc. Natl. Acad. Sci. U.S.A. 105, 20227 (2008)]. Thereby, generalized ensemble simulations can simultaneously escape both the explicit barriers along the collective variable direction and the hidden barriers that are strongly coupled with the collective variable move. As demonstrated in our model studies, the present OSRW based generalized ensemble treatments show improved sampling capability over the corresponding classical generalized ensemble treatments. PMID- 19548710 TI - Effective atomic orbitals for fuzzy atoms. AB - The method of extracting effective atomic orbitals and effective minimal basis sets from molecular wave function characterizing the state of an atom in a molecule is developed in the framework of the "fuzzy" atoms. In all cases studied, there were as many effective orbitals that have considerable occupation numbers as orbitals in the classical minimal basis. That is considered to be of high conceptual importance. PMID- 19548711 TI - The influence of dissipation on the quantum-classical correspondence: stability of stochastic trajectories. AB - The quantum-classical correspondence in the presence of dissipation is studied. The semiclassical expression for the linear response function of an anharmonic system is expressed in a series containing classical stability matrix elements, which can diverge due to the chaotic behavior of stochastic trajectories. The presence of dissipation in most cases removes the divergence of higher-order correction terms, thus suppressing quantum effects and making the system more classical. The regime of system-bath coupling, which makes quantum dynamics completely classical, is obtained in terms of friction, temperature, and anharmonicity. Special cases when bath coupling may lead to enhancement of quantum effects are discussed. PMID- 19548712 TI - On the determination of optimized, fully quadratic, coupled state quasidiabatic Hamiltonians for determining bound state vibronic spectra. AB - The quasidiabatic, coupled electronic state, fully quadratic Hamiltonian (H(d)), suitable for the simulation of spectra exhibiting strong vibronic couplings and constructed using a recently introduced pseudonormal equations approach, is studied. The flexibility inherent in the normal equations approach is shown to provide a robust means for (i) improving the accuracy of H(d), (ii) extending its domain of utility, and (iii) determining the limits of the fully quadratic model. The two lowest electronic states of pyrrolyl which are coupled by conical intersections are used as a test case. The requisite ab initio data are obtained from large multireference configuration interaction expansions comprised of 108.5x10(6) configuration state functions and based on polarized triple zeta quality atomic orbital bases. PMID- 19548713 TI - Harmonic oscillator in presence of nonequilibrium environment. AB - Based on a microscopic Hamiltonian picture where the system is coupled with the nonequilibrium environment, comprising of a set of harmonic oscillators, the Langevin equation with proper microscopic specification of Langevin force is formulated analytically. In our case, the reservoir is perturbed by an external force, either executing rapid or showing periodic fluctuations, hence the reservoir is not in thermal equilibrium. In the presence of external fluctuating force, using Shapiro-Loginov procedure, we arrive at the linear coupled first order differential equations for the two-time correlations and examine the time evolution of the same considering the system as a simple harmonic oscillator. We study the stochastic resonance phenomena of a Kubo-type oscillator (assumed to be the system) when the bath is modulated by a periodic force. The result(s) obtained here is of general significance and can be used to analyze the signature of stochastic resonance. PMID- 19548715 TI - Unified treatment of quantum coherent and incoherent hopping dynamics in electronic energy transfer: reduced hierarchy equation approach. AB - A new quantum dynamic equation for excitation energy transfer is developed which can describe quantum coherent wavelike motion and incoherent hopping in a unified manner. The developed equation reduces to the conventional Redfield theory and Forster theory in their respective limits of validity. In the regime of coherent wavelike motion, the equation predicts several times longer lifetime of electronic coherence between chromophores than does the conventional Redfield equation. Furthermore, we show quantum coherent motion can be observed even when reorganization energy is large in comparison to intersite electronic coupling (the Forster incoherent regime). In the region of small reorganization energy, slow fluctuation sustains longer-lived coherent oscillation, whereas the Markov approximation in the Redfield framework causes infinitely fast fluctuation and then collapses the quantum coherence. In the region of large reorganization energy, sluggish dissipation of reorganization energy increases the time electronic excitation stays above an energy barrier separating chromophores and thus prolongs delocalization over the chromophores. PMID- 19548714 TI - On the adequacy of the Redfield equation and related approaches to the study of quantum dynamics in electronic energy transfer. AB - The observation of long-lived electronic coherence in photosynthetic excitation energy transfer (EET) by Engel et al. [Nature (London) 446, 782 (2007)] raises questions about the role of the protein environment in protecting this coherence and the significance of the quantum coherence in light harvesting efficiency. In this paper we explore the applicability of the Redfield equation in its full form, in the secular approximation and with neglect of the imaginary part of the relaxation terms for the study of these phenomena. We find that none of the methods can give a reliable picture of the role of the environment in photosynthetic EET. In particular the popular secular approximation (or the corresponding Lindblad equation) produces anomalous behavior in the incoherent transfer region leading to overestimation of the contribution of environment assisted transfer. The full Redfield expression on the other hand produces environment-independent dynamics in the large reorganization energy region. A companion paper presents an improved approach, which corrects these deficiencies [A. Ishizaki and G. R. Fleming, J. Chem. Phys. 130, 234111 (2009)]. PMID- 19548716 TI - Development and assessment of a short-range meta-GGA functional. AB - Short-range DFT/long-range ab initio methods allow for a combination of the weak basis-set dependency of DFT with an accurate ab initio treatment of long-range effects like van der Waals interaction. In order to improve existing short-range LDA and GGA density functionals, we developed a TPSS-like short-range meta-GGA exchange-correlation functional and checked its performance in long-range CCSD(T) calculations for thermodynamical properties of the G2 set of molecules. PMID- 19548717 TI - Multiple coherent states for first-principles semiclassical initial value representation molecular dynamics. AB - A multiple coherent states implementation of the semiclassical approximation is introduced and employed to obtain the power spectra with a few classical trajectories. The method is integrated with the time-averaging semiclassical initial value representation to successfully reproduce anharmonicity and Fermi resonance splittings at a level of accuracy comparable to semiclassical simulations of thousands of trajectories. The method is tested on two different model systems with analytical potentials and implemented in conjunction with the first-principles molecular dynamics scheme to obtain the power spectrum for the carbon dioxide molecule. PMID- 19548718 TI - High-performance ab initio density matrix renormalization group method: applicability to large-scale multireference problems for metal compounds. AB - This article presents an efficient and parallelized implementation of the density matrix renormalization group (DMRG) algorithm for quantum chemistry calculations. The DMRG method as a large-scale multireference electronic structure model is by nature particularly efficient for one-dimensionally correlated systems, while the present development is oriented toward applications for polynuclear transition metal compounds, in which the macroscopic one-dimensional structure of electron correlation is absent. A straightforward extension of the DMRG algorithm is proposed with further improvements and aggressive optimizations to allow its application with large multireference active space, which is often demanded for metal compound calculations. Special efficiency is achieved by making better use of sparsity and symmetry in the operator and wave function representations. By accomplishing computationally intensive DMRG calculations, the authors have found that a large number of renormalized basis states are required to represent high entanglement of the electron correlation for metal compound applications, and it is crucial to adopt auxiliary perturbative correction to the projected density matrix during the DMRG sweep optimization in order to attain proper convergence to the solution. Potential energy curve calculations for the Cr(2) molecule near the known equilibrium precisely predicted the full configuration interaction energies with a correlation space of 24 electrons in 30 orbitals [denoted by (24e,30o)]. The energies are demonstrated to be accurate to 0.6mE(h) (the error from the extrapolated best value) when as many as 10,000 renormalized basis states are employed for the left and right DMRG block representations. The relative energy curves for [Cu(2)O(2)](2+) along the isomerization coordinate were obtained from DMRG and other correlated calculations, for which a fairly large orbital space (32e,62o) is modeled as a full correlation space. The DMRG prediction nearly overlaps with the energy curve from the coupled cluster with singles, doubles, and perturbative triple [CCSD(T)] calculations, while the multireference complete active space self-consistent field (CASSCF) calculations with the small reference configuration (8e,8o) are found to overestimate the biradical character of the electronic state of [Cu(2)O(2)](2+) according to the one-electron density matrix analysis. PMID- 19548719 TI - Car-Parrinello density matrix search with a first principles fictitious electron mass method for electronic wave function optimization. AB - In spite of its success in molecular dynamics and the advantage of being a first order propagation technique, the Car-Parrinello method and its variations have not been successful in self-consistent-field (SCF) wave function optimization due to slow convergence. In this article, we introduce a first principles fictitious mass scheme to weigh each individual density element differently and instantaneously. As an alternative to diagonalization in SCF, the Car-Parrinello scheme is implemented as a density matrix search method. Not only does the fictitious mass scheme developed herein allow a very fast SCF convergence, but also the Car-Parrinello density matrix search (CP-DMS) exhibits linear scaling with respect to the system size for alanine helical chain test molecules. The excellent performance of CP-DMS holds even for very challenging compact three dimensional quantum particles. While the conventional diagonalization based SCF method has difficulties optimizing electronic wave functions for CdSe quantum dots, CP-DMS shows both smooth and faster convergence. PMID- 19548720 TI - Measurement of work in single-molecule pulling experiments. AB - A main goal of single-molecule experiments is to evaluate equilibrium free energy differences by applying fluctuation relations to repeated work measurements along irreversible processes. We quantify the error that is made in a free energy estimate by means of the Jarzynski equality when the accumulated work expended on the whole system (including the instrument) is erroneously replaced by the work transferred to the subsystem consisting of the sole molecular construct. We find that the error may be as large as 100%, depending on the number of experiments and on the bandwidth of the data acquisition apparatus. Our theoretical estimate is validated by numerical simulations and pulling experiments on DNA hairpins using optical tweezers. PMID- 19548721 TI - A reanalysis of the A 1A"-X 1A' transition of CFBr. AB - The laser induced fluorescence spectrum of the A (1)A(")-X (1)A(') transition of CFBr is presented, with selected bands recorded at sub-Doppler resolution, allowing the rotational constants to be fully determined. Analysis of dispersed fluorescence spectra and the pattern of (79)Br/(81)Br isotope splittings indicate that the origin must be shifted from previous assignments in the literature to 23 271.0 cm(-1). This implies that only the lowest four vibrational levels in the A state have significant quantum yields for fluorescence, with all other levels strongly predissociated. Comparison with photofragment measurements implies that the A state is metastable, with a barrier to dissociation of approximately 1000 cm(-1). PMID- 19548722 TI - Symmetry-adapted density matrix renormalization group calculations of the primary excited states of poly(para-phenylene vinylene). AB - The Pariser-Parr-Pople model of pi-conjugated electrons is solved by a three block, symmetry-adapted density matrix renormalization group (DMRG) method for the light emitting polymer, poly(para-phenylene vinylene). The energies of the primary excited states are calculated. There is excellent agreement between theory and experiment when solid state screening is incorporated into the model parameters, enabling us to make an identification of the origin of the key spectroscopic features. Appendices describe important technical aspects of the three-block DMRG approach: Local Hilbert space efficiency and its relation to the matrix product formulation of the DMRG; an efficient computational procedure for constructing symmetry-adapted states for DMRG calculations; and correct superblock state targeting to ensure good convergence of the method. PMID- 19548723 TI - Coupled-surface investigation of the photodissociation of NH3(A): effect of exciting the symmetric and antisymmetric stretching modes. AB - Using previously developed potential energy surfaces and their couplings, non Born-Oppenheimer trajectory methods are used to study the state-selected photodissociation of ammonia, prepared with up to six quanta of vibrational excitation in the symmetric (nu(1)) or antisymmetric (nu(3)) stretching modes of NH(3)(A). The predicted dynamics is mainly electronically nonadiabatic (that is, it produces ground electronic state amino radicals). The small probability of forming the excited-state amino radical is found, for low excitations, to increase with total energy and to be independent of whether the symmetric or antisymmetric stretch is excited; however some selectivity with respect to exciting the antisymmetric stretch is found when more than one quantum of excitation is added to the stretches, and more than 50% of the amino radical are found to be electronically excited when six quanta are placed in the antisymmetric stretch. These results are in contrast to the mechanism inferred in recent experimental work, where excitation of the antisymmetric stretch by a single quantum was found to produce significant amounts of excited-state products via adiabatic dissociation at total energies of about 7.0 eV. Both theory and experiment predict a broad range of translational energies for the departing H atoms when the symmetric stretch is excited, but the present simulations do not reproduce the experimental translational energy profiles when the antisymmetric stretch is excited. The sensitivity of the predicted results to several aspects of the calculation is considered in detail, and the analysis leads to insight into the nature of the dynamics that is responsible for mode selectivity. PMID- 19548724 TI - A laboratory and theoretical study of protonated carbon disulfide, HSCS+. AB - The rotational spectrum of protonated carbon disulfide, HSCS(+), has been detected in the centimeter-wave band in a molecular beam by Fourier transform microwave spectroscopy. Rotational and centrifugal distortion constants have been determined from ten transitions in the K(a)=0 ladder of the normal isotopic species, HS(13)CS(+), and DSCS(+). The present assignment agrees well with high level coupled cluster calculations of the HSCS(+) structure, which, like earlier work, predict this isomer to be the ground state on the HCS(2) (+) potential energy surface; HCSS(+), an isomer with C(2v) symmetry, is predicted to lie more than 20 kcal/mol higher in energy. Other properties of HSCS(+) including its dipole moment, anharmonic vibrational frequencies, and infrared intensities have also been computed at the coupled cluster level of theory with large basis sets. Because carbon disulfide possesses a fairly large proton affinity, and because this nonpolar molecule may plausibly exist in astronomical sources, HSCS(+) is a good candidate for detection with radio telescopes in the submillimeter band where the stronger b-type transitions of this protonated cation are predicted to lie. PMID- 19548725 TI - Full dimensional (15-dimensional) quantum-dynamical simulation of the protonated water-dimer III: Mixed Jacobi-valence parametrization and benchmark results for the zero point energy, vibrationally excited states, and infrared spectrum. AB - Quantum dynamical calculations are reported for the zero point energy, several low-lying vibrational states, and the infrared spectrum of the H(5)O(2)(+) cation. The calculations are performed by the multiconfiguration time-dependent Hartree (MCTDH) method. A new vector parametrization based on a mixed Jacobi valence description of the system is presented. With this parametrization the potential energy surface coupling is reduced with respect to a full Jacobi description, providing a better convergence of the n-mode representation of the potential. However, new coupling terms appear in the kinetic energy operator. These terms are derived and discussed. A mode-combination scheme based on six combined coordinates is used, and the representation of the 15-dimensional potential in terms of a six-combined mode cluster expansion including up to some 7-dimensional grids is discussed. A statistical analysis of the accuracy of the n mode representation of the potential at all orders is performed. Benchmark, fully converged results are reported for the zero point energy, which lie within the statistical uncertainty of the reference diffusion Monte Carlo result for this system. Some low-lying vibrationally excited eigenstates are computed by block improved relaxation, illustrating the applicability of the approach to large systems. Benchmark calculations of the linear infrared spectrum are provided, and convergence with increasing size of the time-dependent basis and as a function of the order of the n-mode representation is studied. The calculations presented here make use of recent developments in the parallel version of the MCTDH code, which are briefly discussed. We also show that the infrared spectrum can be computed, to a very good approximation, within D(2d) symmetry, instead of the G(16) symmetry used before, in which the complete rotation of one water molecule with respect to the other is allowed, thus simplifying the dynamical problem. PMID- 19548726 TI - Photodissociation dynamics of IrBr6(2-) dianions by time-resolved photoelectron spectroscopy. AB - We have used femtosecond time-resolved photoelectron spectroscopy to examine the photodissociation dynamics of doubly charged anions IrBr(6)(2-) after excitation at h nu(pump) = 1.6 eV and with a detachment photon energy of h nu(probe) = 4.8 eV. Excited state dynamics proceed by successive decay of the initially excited state, by way of an intermediate and back to the electronic ground state. This is associated with lifetimes of tau(1) = 2.1+/-0.3 ps and tau(2) = 21+/-2 ps, respectively. After nonadiabatic relaxation, the internal energy of the dianion is sufficiently large to induce fragmentation and delayed emission of Br(-) over the repulsive Coulomb barrier with a 79+/-21 ps time constant. As both fragments are negatively charged, Coulomb repulsion at early times (and correspondingly small separations) is reflected in the transient photoelectron spectra. Analysis of both shifts and intensities of the time-dependent bromide detachment features allows determination of the shape of the dissociation barrier. A lower limit of the outer height was retrieved from the kinetic energy release of KER > or = 1.6 eV. Modeling of the dissociation rate with statistical rate theory results in an inner barrier height of E(RCB) = 0.95 eV. PMID- 19548727 TI - Study of the interaction between water and hydrogen sulfide with polycyclic aromatic hydrocarbons. AB - A computational study has been carried out for determining the characteristics of the interaction between one water and hydrogen sulfide molecule with a series of polycyclic aromatic hydrocarbons of increasing size, namely, benzene, anthracene, triphenylene, coronene, circumcoronene, and dicircumcoronene. Potential energy curves were calculated for structures where H(2)X (X=O,S) molecule is located over the central six-membered ring with its hydrogen atoms pointing toward to (mode A) or away from (mode B) the hydrocarbon. The accuracy of different methods has been tested against the results of coupled cluster calculations extrapolated to basis set limit for the smaller hydrocarbons. The spin component scaled MP2 (SCS-MP2) method and a density functional theory method empirically corrected for dispersion (DFT-D) reproduce fairly well the results of high level calculations and therefore were employed for studying the larger systems, though DFT-D seems to underestimate the interaction in hydrogen sulfide clusters. Water complexes in mode A have interaction energies that hardly change with the size of the hydrocarbon due to compensation between the increase in the correlation contribution to the interaction energy and the increase in the repulsive character of the Hartree-Fock energy. For all the other clusters studied, there is a continuous increase in the intensity of the interaction as the size of the hydrocarbon increases, suggesting already converged values for circumcoronene. The interaction energy for water clusters extrapolated to an infinite number of carbon atoms amounts to -13.0 and -15.8 kJ/mol with SCS-MP2 and DFT-D, respectively. Hydrogen sulfide interacts more strongly than water with the hydrocarbons studied, leading to a limiting value of -21.7 kJ/mol with the SCS MP2 method. Also, complexes in mode B are less stable than the corresponding A structures, with interaction energies amounting to -8.2 and -18.2 kJ/mol for water and hydrogen sulfide, respectively. The DFT-D calculations give values of 16.2 and -9.3 kJ/mol for hydrogen sulfide complexes in modes A and B, less negative than those predicted by the SCS-MP2 method, probably indicating problems with sulfur dispersion parameters. PMID- 19548728 TI - A crossed beams and ab initio investigation on the formation of cyanodiacetylene in the reaction of cyano radicals with diacetylene. AB - The crossed molecular beams reaction of ground state cyano radicals (CN) with diacetylene (HCCCCH) was studied in the laboratory under single collision conditions. Combining the derived center-of-mass translational energy and angular distributions with novel electronic structure calculations, we show that the linear cyanodiacetylene molecule (HCCCCCN) is the sole reaction product. Our study provided no substantiation of two alternative products which have been suggested previously: cyanoacetylene (HCCCN), speculated to be synthesized via the exchange of the ethynyl by the cyano group, and the 1,3-butadiynyl radical (HCCCC), thought to be formed via hydrogen abstraction. The unambiguous identification of cyanodiacetylene formed in an exoergic, barrierless bimolecular reaction of the cyano radical with diacetylene strongly suggests that cyanodiacetylene can be also synthesized via this process in the interstellar medium (cold molecular clouds) and in hydrocarbon-rich atmospheres of planets and their moons such as Titan. PMID- 19548730 TI - A combined experimental and theoretical study on realizing and using laser controlled torsion of molecules. AB - It is demonstrated that strong laser pulses can introduce torsional motion in the axially chiral molecule 3,5-difluoro-3('),5(')-dibromobiphenyl. A nanosecond laser pulse spatially aligns the stereogenic carbon-carbon (C-C) bond axis allowing a perpendicularly polarized, intense femtosecond pulse to initiate torsional motion accompanied by a rotation about the fixed axis. We monitor the induced motion by femtosecond time-resolved Coulomb explosion imaging. Our theoretical analysis corroborates the experimental findings and on the basis of these results we discuss future applications of laser-induced torsion, viz., time resolved studies of deracemization and laser controlled molecular junctions based on molecules with torsion. PMID- 19548729 TI - Oxidation of Al doped Au clusters: a first principles study. AB - Using first principles method we report the oxidation of Al doped Au clusters. This work is divided into two parts: (i) the equilibrium structures and stability of Al doped Au(n-1) clusters (n=2-7,21) and (ii) the interaction of O(2) with stable clusters. The calculations are performed using the plane wave pseudopotential approach under the density functional theory and generalized gradient approximation for the exchange and correlation functional. The optimized geometries of Au(n-1)Al clusters indicate that the substitution of Au by Al results an early onset of three-dimensional structures from tetramer onwards. This is different from the results of transition metal doped Au clusters, where the planar conformation of Au clusters retains up to heptamer. The stability of Au(n-1)Al clusters has been analyzed based on the binding energy, second difference in energy, and the energy gaps between the highest occupied molecular orbital and lowest unoccupied molecular orbital energy levels. Based on the energetics, the Au(3)Al and Au(5)Al clusters are found to have extraordinary stability. The oxidation mechanism of Al doped Au clusters have been studied by the interaction of O(2) with Al, Au, AuAl, Au(3)Al, and Au(20)Al clusters. It is found that the oxidation of Au(n-1)Al clusters undergoes via dissociative mechanism, albeit significant charge transfer from Al to Au. Moreover, the O(2) molecule prefers to attach at the Al site rather than at the Au site. PMID- 19548731 TI - OH produced from o-nitrophenol photolysis: a combined experimental and theoretical investigation. AB - Photodissociation dynamics of o-nitrophenol in the gas phase at different photolysis wavelengths (361-390 nm) is investigated, and the nascent OH radical is observed by the single-photon laser-induced fluorescence technique. At all the photolysis wavelengths, the OH radicals are formed in vibrationally cold state (upsilon(")=0) and have similar rotational state distributions. The average rotational temperature for all the photolysis wavelengths is approximately 970+/ 120 K, corresponding to a rotational energy of 1.9+/-0.2 kcal mol(-1). The spin orbit and Lambda-doublet states of the OH fragments formed in the dissociation are measured to have nonstatistical distributions. To get an insight into the dissociative mechanism leading to OH formation in the photolysis of o nitrophenol, the potential energy surfaces of the OH-forming channels are mapped by ab initio theoretical calculations. According to both experimental and theoretical results, a possible mechanism for OH formation is proposed. PMID- 19548732 TI - Anomalous enrichment of 17O and 13C in photodissociation products of CO2: possible role of nuclear spin. AB - Oxygen and carbon isotope fractionation associated with products (CO and O(2)) of gas phase photodissociation of CO(2) have been studied using photons from Hg lamp (184.9 nm) and Kr lamp (123.6 and 116.5 nm). In dissociation by Hg lamp photons both CO and O(2) are enriched in (17)O by about 81 per thousand compared to the estimate based on a kinetic model. Additionally, CO is enriched in (13)C by about 37 per thousand relative to the model composition. In contrast, in dissociation by higher energy Kr lamp photons no such anomaly was found in O(2). The observed isotopic enrichments in case of Hg lamp dissociation are proposed to be due to a hyperfine interaction between nuclear spin and electron spins or orbital motion causing enhanced dissociation of isotopologues of CO(2) containing (17)O and (13)C. The (17)O enrichment is higher than that of (13)C by a factor of 2.2+/-0.2 which can be explained by the known magnetic moment ratio of (17)O and (13)C due to differing nuclear spins and g-factors. These results have potential implications in studies of the planetary atmospheres. PMID- 19548733 TI - Structure and dynamics of a Gay-Berne liquid crystal confined in cylindrical nanopores. AB - Gay-Berne liquid crystals confined in two cylindrical nanopores with different pore sizes were studied by molecular dynamics simulation. Their structure and dynamics properties were obtained and compared with those of the bulk. Our data show that confinement changes the bulk isotropic-to-nematic transition to a continuous ordering from a paranematic to a nematic phase. Moreover, confinement strongly hinders the smectic translational order. The molecular dynamics is characterized by the translational diffusion coefficients and the first-rank reorientational correlation times. Very different characteristic times and temperature variations in the dynamics are observed in confinement. Spatially resolved quantities illustrate that confinement induces predominant structural and dynamical heterogeneities. PMID- 19548735 TI - Composition dependence of glass transition temperature and fragility. II. A topological model of alkali borate liquids. AB - Glass transition temperature and fragility are two important properties derived from the temperature dependence of the shear viscosity of glass-forming melts. While direct calculation of these properties from atomistic simulations is currently infeasible, we have developed a new topological modeling approach that enables accurate prediction of the scaling of both glass transition temperature and fragility with composition. A key feature of our approach is the incorporation of temperature-dependent constraints that become rigid as a liquid is cooled. Using this approach, we derive analytical expressions for the composition (x) dependence of glass transition temperature, T(g)(x), and fragility, m(x), in binary alkali borate systems. Results for sodium borate and lithium borate systems are in agreement with published values of T(g)(x) and m(x). Our modeling approach reveals a natural explanation for the presence of the constant T(g) regime observed in alkali borate systems. PMID- 19548734 TI - Hydrogen transfer and hydration properties of H(n)PO4(3-n) (n=0-3) in water studied by first principles molecular dynamics simulations. AB - Density functional theory Perdew-Burke-Ernzerhof [Perdew et al., Phys. Rev. Lett. 77, 3865 (1996)] molecular dynamics simulations of aqueous solutions of orthophosphate species H(n)PO(4)(3-n) (n=0-3) provide new insights into hydrogen transfer and intermolecular and hydration properties of these important aqueous species. Extensive Car-Parrinello molecular dynamics simulations of the orthophosphate ion PO(4)(3-), of the hydrogen phosphate anions, HPO(4)(2-) and H(2)PO(4)(-), and of the orthophosphoric acid, H(3)PO(4), in explicit water show that the process of proton transfer from H(n)PO(4)(3-n) to the surrounding water molecules is very fast, less than 1 ps, and indicate that the dehydrogenation occurs through a concerted proton hopping mechanism, which involves H(n)PO(4)(3 n) and three water molecules. Analysis of the intermolecular H(n)PO(4)(3-n)-water structure shows that the PO(4)(3-) anions have a significant effect on the H bonding network of bulk water and the presence of P-O(-) moieties induce the formation of new types of H-H interactions around this orthophosphate. Calculated probability distributions of the coordination numbers of the first hydration shell of PO(4)(3-), HPO(4)(2-), and H(2)PO(4)(-) show that these phosphate species display a flexible first coordination shell (between 7 and 13 water molecules) and that the flexibility increases on going from PO(4)(3-) to H(2)PO(4)(-). The strength and number of hydrogen bonds of PO(4)(3-), HPO(4)(2-), and H(2)PO(4)(-) are determined through a detailed analysis of the structural correlation functions. In particular, the H-bond interactions between the oxygen atoms of the phosphates and the surrounding water molecules, which decrease on going from PO(4)(3-) to the hydrogenated H(2)PO(4)(-) species, explain the diminished effect on the structure of water with the increasing hydrogenation of the orthophosphate anions. PMID- 19548736 TI - Static dielectric properties of polarizable ion models: molecular dynamics study of molten AgI and NaI. AB - The fluctuation-dissipation theorem for the static dielectric response function of systems of ions with inducible point dipoles is derived. It is shown that the static longitudinal dielectric function is determined by spatial correlations of both charge and dipole-moment density fluctuations. Moreover, it is deduced that the long-wavelength behavior of the charge structure factor for polarizable ion systems is different from that for systems of rigid ions. Molecular dynamics simulation results of rigid and polarizable ion models for molten AgI and NaI are reported. PMID- 19548737 TI - Characterization of transient molecular vibration excited with shaped femtosecond pulses. AB - We study vibrational dynamics of molecules interacting with spectrally shaped broadband laser pulses. After performing a single measurement based on cross correlation frequency resolved optical gating of molecular vibration, complete evolution of the complex-valued quantum coherence between the vibrational states is reconstructed with variable time and frequency resolution. The ability to change the resolution in the analysis of the transient molecular dynamics without repeating the experiment or changing experimental parameters is useful in designing and understanding various schemes of controlling quantum states of molecules. PMID- 19548738 TI - Width of reaction zones in A+B-->C type reaction-diffusion processes: effects of an electric current. AB - We investigate the effects of an electric current on the width of a stationary reaction zone in an irreversible A(-)+B(+)-->C reaction-diffusion process. The ion dynamics of electrolytes A identical with (A(+),A(-)) and B identical with (B(+),B(-)) is described by reaction-diffusion equations obeying local electroneutrality and the stationary state is obtained by employing reservoirs of fixed electrolyte concentrations at the opposite ends of a finite domain. We find that the width of the reaction zone decreases when the current drives the reacting ions toward the reaction zone while it increases in the opposite case. The linear response of the width to the current is estimated by developing a phenomenological theory based on conservation laws and on electroneutrality. The theory is found to reproduce numerical solutions to a good accuracy. PMID- 19548739 TI - Effect of the static magnetic field strength on parahydrogen induced polarization NMR spectra. AB - Spin polarization transfer from parahydrogen (p-H(2)) to another molecular entity is generally thought to be mediated by longitudinal spin order (represented by the operator product I(z)(A)I(z)(B), A and B being the two hydrogen nuclei which originate from p-H(2) after a hydrogenation reaction). The longitudinal spin order leads to antiphase patterns in the proton NMR spectrum. In addition to these antiphase patterns, in-phase patterns, arising from polarization differences (represented by (I(z)(A)-I(z)(B))), have been experimentally observed. A complete theory, based on a density operator treatment, has been worked out and applied to the two types of parahydrogen induced polarization experiments: PASADENA (PArahydrogen and Synthesis Allow Dramatically Enhanced Nuclear Alignment; hydrogenation reaction inside the NMR magnet) and (ALTADENA) (Adiabatic Longitudinal Transport After Dissociation Engenders Nuclear Alignment; hydrogenation reaction outside the NMR magnet). It is shown that polarization differences are always created in the case of a PASADENA experiment but that their amplitude depends critically on the ratio of the J coupling over the frequency difference between A and B. In the case of an ALTADENA experiment, if the sample is slowly transferred toward the NMR magnet, polarization differences are definitely created and their amplitude can be larger than the amplitude of the longitudinal spin order. Some test experiments demonstrate the validity of the proposed theory. PMID- 19548741 TI - Density functional theory study on Herzberg-Teller contribution in Raman scattering from 4-aminothiophenol-metal complex and metal-4-aminothiophenol-metal junction. AB - Density functional theory (DFT) and time-dependent DFT calculations have been performed to investigate the Raman scattering spectra of metal-molecule complex and metal-molecule-metal junction architectures interconnected with 4 aminothiophenol (PATP) molecule. The simulated profiles of normal Raman scattering (NRS) spectra for the two complexes (Ag(2)-PATP and PATP-Au(2)) and the two junctions (Ag(2)-PATP-Au(2) and Au(2)-PATP-Ag(2)) are similar to each other, but exhibit obviously different Raman intensities. Due to the lager static polarizabilities of the two junctions, which directly influence the ground state chemical enhancement in NRS spectra, the calculated normal Raman intensities of them are stronger than those of two complexes by the factor of 10(2). We calculate preresonance Raman scattering (RRS) spectra with incident light at 1064 nm, which is much lower than the S(1) electronic transition energy of complexes and junctions. Ag(2)-PATP-Au(2) and Au(2)-PATP-Ag(2) junctions yield higher Raman intensities than those of Ag(2)-PATP and PATP-Au(2) complexes, especially for b(2) modes. This effect is mainly attributed to charge transfer (CT) between the metal gap and the PAPT molecule which results in the occurrence of CT resonance enhancement. The calculated pre-RRS spectra strongly depend on the electronic transition state produced by new structures. With excitation at 514.5 nm, the calculated pre-RRS spectra of two complexes and two junctions are stronger than those of with excitation at 1064 nm. A charge difference densities methodology has been used to visually describe chemical enhancement mechanism of RRS spectrum. This methodology aims at visualizing intermolecular CT which provides direct evidence of the Herzberg-Teller mechanism. PMID- 19548742 TI - Photoabsorption cross section of C70 thin films from visible to vacuum ultraviolet. AB - Absolute photoabsorption cross sections of C(70) thin films were determined for hv values from 1.3 to 42 eV using photon attenuation. The spectrum showed a prominent peak of 1320 Mb at 21.4 eV with several fine structures mostly due to sigma-->sigma(*) single-electron excitation. The complex refractive index and complex dielectric function were calculated up to 42 eV with Kramers-Kronig analyses. From the present data of C(70) thin films, the cross section curve of molecular C(70) was calculated using the standard Clausius-Mossotti relation dealing with correction of the local electromagnetic field, with a plausible assumption that the anisotropy in molecular structure of C(70) was smeared out by molecular rotation at room temperature. PMID- 19548740 TI - Concentration fluctuations in fluid mixtures. II. AB - The method developed in a previous paper [R. Mazo, J. Chem. Phys. 129, 154101 (2008)] for extracting information on the size of relative fluctuations in multicomponent systems is tested on ten binary systems and one ternary system. For the binary systems, it is found that the approximation works well for mole fractions in the range of 0.15-0.85 in most cases. For the ternary case, the method inherently yields less information and is valid only over a more restricted range for the case studied (chloroform-methanol-acetone). It is found that the predicted ratio of number fluctuations is approximately equal to the ratio of molar volumes of the components. PMID- 19548743 TI - Direct correlation function of the square-well fluid with attractive well width up to two particle diameters. AB - Analytical expression for direct correlation function of the square-well fluid with an attractive well width up to two particle diameters (2 < lambda < or = 3) is reported. This result is obtained within the first-order mean-spherical approximation (FMSA) and represents the nontrivial extension of the recent study due to Tang [J. Chem. Phys. 127, 164504 (2007)], where the width of square-well attraction was limited by one particle diameter (1 < lambda < or = 2). Prediction of the FMSA theory is validated by direct comparison against Monte Carlo simulation data. Additionally, an impact of the increase in the range of attraction on the parameters of the critical point of the square-well fluid is discussed using the compressibility route to thermodynamics. PMID- 19548744 TI - Exciton-phonon coupling in molecular crystals: synergy between two intramolecular vibrational modes in quaterthiophene single crystals. AB - Exciton-phonon (EP) coupling in molecular crystals is investigated in the case where two intramolecular vibrational modes are involved and a theoretical model is presented which applies when one of the modes is strongly coupled to crystal excitons. The model is used to simulate the low energy portion of the absorption spectra of quaterthiophene (4T) single crystals, for which we find it appropriate to consider a low energy vibrational mode at 161 cm(-1) and an effective strongly coupled high energy mode at 1470 cm(-1). Our numerical results demonstrate that the high energy mode renormalizes the excitonic band, thereby strongly affecting the environment seen by the low energy mode and the overall EP coupling regime. Numerical simulations also confirm the existence of the new coupling regimes "intermediate-I" and "strong-I" already introduced for oligothiophene aggregates [Spano et al., J. Chem. Phys. 127, 184703 (2007)], which arise as a consequence of the large effective mass of low energy excitons in 4T crystals. Comparison with experimental high resolution absorption spectra is also reported and shown to support the model predictions. PMID- 19548745 TI - Interfacial water: a first principles molecular dynamics study of a nanoscale water film on salt. AB - Density functional theory (DFT) molecular dynamics simulations of a thin (approximately 15 A) water film on NaCl(001) have been performed, with the aim of understanding the structural and dynamic properties of this important interfacial water system. The interaction of the water film with the surface orders the water molecules in the immediate vicinity of the interface. This is reflected by oscillations in the planar-averaged water density distribution along the surface normal that extend to about 8 A from the surface. The interaction with the substrate leaves many of the water molecules in the immediate vicinity with broken hydrogen bonds and as a consequence considerably reduced dipole moments. Indeed a clear correlation between the number of hydrogen bonds which a water molecule is involved in and its dipole moment for both water on NaCl and bulk water is observed. How the DFT results obtained here compare to those obtained with various empirical potentials is briefly discussed. PMID- 19548746 TI - The preserved aromaticity of aniline molecules adsorbed on a Si(5 5 12)-2x1 surface. AB - We present a scanning tunneling microscopy and first-principles calculations study of the adsorption structures of aniline on a Si(5 5 12)-2x1 surface. Dissociation from the aniline molecules of one or two H atom(s) bonded to N is favored, and then adsorption onto adatom, tetramer, and dimer rows of Si(5 5 12) 2x1 occurs in several distinct configurations. On the adatom row, aniline binds to an adatom in a tilted configuration, which is formed via a sigma bond between the adatom and N, with one dissociated H atom adsorbed on a nearby adatom. No further hydrogen dissociation occurs. On the tetramer and dimer rows, the structures with two dissociated hydrogens and upright configurations are the most stable. Aniline does not adsorb onto the honeycomb chains; this adsorption configuration has a low adsorption energy. In all the adsorption configurations of aniline on this surface, the molecule's aromaticity is preserved. Thus Si-N bonding of aromatic amine molecules provides a strategy for the homogeneous aromatic functionalization of high index Si surfaces. PMID- 19548747 TI - Standard electrode potential, Tafel equation, and the solvation thermodynamics. AB - Equilibrium in the electronic subsystem across the solution-metal interface is considered to connect the standard electrode potential to the statistics of localized electronic states in solution. We argue that a correct derivation of the Nernst equation for the electrode potential requires a careful separation of the relevant time scales. An equation for the standard metal potential is derived linking it to the thermodynamics of solvation. The Anderson-Newns model for electronic delocalization between the solution and the electrode is combined with a bilinear model of solute-solvent coupling introducing nonlinear solvation into the theory of heterogeneous electron transfer. We therefore are capable of addressing the question of how nonlinear solvation affects electrochemical observables. The transfer coefficient of electrode kinetics is shown to be equal to the derivative of the free energy, or generalized force, required to shift the unoccupied electronic level in the bulk. The transfer coefficient thus directly quantifies the extent of nonlinear solvation of the redox couple. The current model allows the transfer coefficient to deviate from the value of 0.5 of the linear solvation models at zero electrode overpotential. The electrode current curves become asymmetric in respect to the change in the sign of the electrode overpotential. PMID- 19548748 TI - Surface enhanced Raman scattering of pyridine adsorbed on Au@Pd core/shell nanoparticles. AB - Surface enhanced Raman scattering (SERS) of pyridine adsorbed on Au@Pd core/shell nanoparticles has been investigated theoretically with quantum chemical method, generalized Mie theory and three-dimensional finite-difference time domain (3D FDTD) method. We first studied the influence of the coated Pd on the electronic structure of Au nanoparticle, and compared the electronic structure of Au(20) cluster with that of Au(10)Pd(10) (core/shell) cluster. Second, we studied SERS spectroscopy of pyridine on Au@Pd core/shell nanoparticles, which revealed the rate of static chemical enhancement and electromagnetic enhancement in the experimental reports. Third, the influence of the Pd shell thickness to the optical absorption of Au@Pd core/shell nanoparticles was investigated with generalized Mie theory. Fourth, we studied the influence of the shell thickness to the local electric field enhancement with 3D-FDTD method. The theoretical results reveal that the static chemical enhancement and electromagnetic enhancement are in the order of 10 and 10(3), respectively. These theoretical studies promote the deeper understanding of the electronic structure and optical absorption properties of Au@Pd, and the mechanisms for SERS of molecule adsorbed on Au@Pd. PMID- 19548749 TI - Endohedral nitrogen storage in carbon fullerene structures: physisorption to chemisorption transition with increasing gas pressure. AB - We present extensive pseudopotential density functional theory (DFT) calculations in order to analyze the structural properties and chemical reactivity of nitrogen molecules confined in spheroidal (C(82)) and tubelike (C(110)) carbon fullerene structures. For a small number of encapsulated nitrogens, the N(2) species exist in a nonbonded state within the cavities and form well defined molecular conformations such as linear chains, zigzag arrays, as well as both spheroidal and tubular configurations. However, with increasing the number of stored molecules, the interaction among the confined nitrogens as well as between the N(2) species and the fullerene wall is not always mainly repulsive. Actually, at high densities of the encapsulated gas, we found both adsorption of N(2) to the inner carbon surface together with the formation of (N(2))(m) molecular clusters. Total energy DFT calculations reveal that the shape of the interaction potential of a test molecule moving within the carbon cavities strongly varies with the number and proximity of the coadsorbed N(2) from being purely repulsive to having short-range attractive contributions close to the inner wall. In particular, the latter are always found when a group of closely spaced nitrogens is located near the carbon cage (a fact that will naturally occur at high densities of the encapsulated gas), inducing the formation of covalent bonds between the N(2) and the fullerene network. Interestingly, in some cases, the previous nitrogen adsorption to the inner surface is reversible by reducing the gas pressure. The calculated average density of states of our considered carbon compounds reveals the appearance of well defined features that clearly reflect the occurring structural changes and modifications in the adsorption properties in the systems. Our results clearly underline the crucial role played by confinement effects on the reactivity of our endohedral compounds, define this kind of materials as nonideal nanocontainers for high density nitrogen storage applications, and must be taken into account when analyzing the diffusion properties of the encapsulated species. PMID- 19548750 TI - Adsorption and pinning of multiblock copolymers on chemically heterogeneous patterned surfaces. AB - The results of Monte Carlo simulations carried out on a system of multiblock copolymers having two different types of monomer units adsorbed on checker board surface configurations are presented here. We investigated the adsorption behavior for a series of different chess board square dimensions and also for various block lengths of the copolymer. We find that the specific heat capacity curves for adsorption indicate double peaks showing a two stage pattern recognition of the copolymer on the surface. It is also seen that the transition that happens at lower temperature corresponds to pinning where the junction point of the different blocks gets pinned to the interface between the different surface sites. It is interesting to see that the multiblock copolymers form multiple pinning sites on the boundary between different kinds of surface sites. There exists an intermediate size of the square on the board where the recognition and pinning are most favored and for smaller and larger size of the board and block length, the adsorption proceeds like in homopolymer on homogeneous surfaces. Unlike in the case of the simplest model of diblock copolymer on stripe-patterned surfaces [K. Sumithra and E. Straube, J. Chem. Phys. 125, 154701 (2006)], here the recognition is stronger and the average adsorption energy and the perpendicular component of the radius of gyration show distinct changes corresponding to the two transitions. The conformational properties of the multiblock copolymer near the checkered surface show interesting effects with the perpendicular component showing strong deviations from the standard behavior. PMID- 19548751 TI - Tethered DNA dynamics in shear flow. AB - We study the cyclic dynamics of a single polymer tethered to a hard wall in shear flow using Brownian dynamics, the lattice Boltzmann method, and a recent stochastic event-driven molecular dynamics algorithm. We focus on the dynamics of the free end (last bead) of the tethered chain and we examine the cross correlation function and power spectral density of the chain extensions in the flow and gradient directions as a function of chain length N and dimensionless shear rate Wi. Extensive simulation results suggest a classical fluctuation dissipation stochastic process and question the existence of periodicity of the cyclic dynamics, as previously claimed. We support our numerical findings with a simple analytical calculation for a harmonic dimer in shear flow. PMID- 19548752 TI - Polydispersity effects in poly(isoprene-b-styrene-b-ethylene oxide) triblock terpolymers. AB - Four hydroxyl-terminated poly(isoprene-b-styrene) diblock copolymers with comparable molecular weights and compositions (equivalent volume fractions of polyisoprene and polystyrene) but different polystyrene block polydispersity indices (M(w)/M(n)=1.06,1.16,1.31,1.44) were synthesized by anionic polymerization using either sec-butyllithium or the functional organolithium 3 triisopropylsilyloxy-1-propyllithium. Poly(ethylene oxide) (PEO) blocks were grown from the end of each of these parent diblocks to yield four series of poly(isoprene-b-styrene-b-ethylene oxide) (ISO) triblock terpolymers that were used to interrogate the effects of varying the polydispersity of the middle bridged polystyrene block. In addition to the neat triblock samples, 13 multicomponent blends were prepared at four different compositions from the ISO materials containing a polystyrene segment with M(w)/M(n)=1.06; these blends were used to probe the effects of increasing the polydispersity of the terminal PEO block. The melt-phase behavior of all samples was characterized using small-angle X-ray scattering and dynamic mechanical spectroscopy. Numerous polydispersity driven morphological transitions are reported, including transitions from lamellae to core-shell gyroid, from core-shell gyroid to hexagonally packed cylinders, and from network morphologies [either O(70) (the orthorhombic Fddd network) or core-shell gyroid] to lamellae. Domain periodicities and order disorder transition temperatures also vary with block polydispersities. Self consistent field theory calculations were performed to supplement the experimental investigations and help elucidate the molecular factors underlying the polydispersity effects. The consequences of varying the polydispersity of the terminal PEO block are comparable to the polydispersity effects previously reported in AB diblock copolymers. Namely, domain periodicities increase with increasing polydispersity and domain interfaces tend to curve toward polydisperse blocks. The changes in phase behavior that are associated with variations in the polydispersity of the middle bridged polystyrene block, however, are not analogous to those reported in AB diblock copolymers, as increases in this middle block polydispersity are not always accompanied by (i) increased domain periodicities and (ii) a tendency for domain interfaces to curve toward the polydisperse domain. These results highlight the utility of polydispersity as a tool to tune the phase behavior of ABC block terpolymers. PMID- 19548753 TI - Microphase separation induced by differential interactions in diblock copolymer/homopolymer blends. AB - Phase behavior of diblock copolymer/homopolymer blends (AB/C) is investigated theoretically. The study focuses on a special case where all three binary pairs, A/B, B/C, and C/A, are miscible. Despite the miscibility of the binary pairs, a closed-loop immiscible region exists in the AB/C blends when the A/C and B/C pair interactions are sufficiently different. Inside the closed-loop, the system undergoes microphase separation, exhibiting different ordered structures. This phenomenon is enhanced when the homopolymer (C) interacts more strongly to one of the blocks (A or B). PMID- 19548754 TI - Shear and extensional deformation of droplets containing polymers and nanoparticles. AB - We investigate the effects of polymer chains and nanoparticles on the deformation of a droplet in shear and extensional flow using computational modeling that accounts for both the solid and fluid phases explicitly. We show that under shear flow, both the nanoparticles and the encapsulated polymers reduce the shear induced deformation of the droplet at intermediate capillary numbers. At high capillary numbers, however, long polymer chains can induce the breakup of the droplet. We find that the latter behavior is dependent on the nature of the imposed flow. Specifically, under extensional flow, long polymers inhibit the droplet breakup and reduce deformation. Overall, the findings provide guidelines for tailoring the stability of filled droplets under an imposed flow, and thus, the results can provide useful design rules in a range of technological applications. PMID- 19548755 TI - Brownian molecular rotors: theoretical design principles and predicted realizations. AB - We propose simple design concepts for molecular rotors driven by Brownian motion and external photochemical switching. Unidirectionality and efficiency of the motion is measured by explicit simulations. Two different molecular scaffolds are shown to yield viable molecular rotors when decorated with suitable substituents. PMID- 19548756 TI - Theoretical study of the effect of ethynyl group on the structure and electrical properties of phenyl-thiadiazole systems as precursors of electron-conducting materials. AB - 2,5-Bis(phenylethynyl)-1,3,4-thiadiazole (PhEtTh) and 2,5-diphenyl-1,3,4 thiadiazole (PhTh) are expected to be building blocks for polymer materials that could be employed to conduct electricity due to their narrow highest occupied molecular orbital-lowest unoccupied molecular orbital (HOMO-LUMO) energy gaps. In this work, a theoretical, comparative study about the effect of the ethynyl group on the planarity and electrical conductivity of this kind of systems has been carried out. Thus, several ab initio (Hartree-Fock, Moller-Plesset) and DFT (B3LYP, B3PW91, M05, M05-2X) methods and basis sets (6-31G(*), 6-31G+G(**), 6 311G(**), cc-pVDZ, cc-pVTZ) have been tested. As a result, PhEtTh showed better properties for its use as electric conducting material relative to PhTh due to its smaller HOMO-LUMO gap, as well as its enhanced trend to retain the planarity provided the reduction in steric hindrances that the ethynyl group (-C[triple bond]C-) permits. Solvent effects were also modeled for ethanol and chloroform under the conductor-like polarizable continuum model approximation. Finally, electronic transitions in gas and solution phases were predicted by using TDDFT approximation in order to compare the theoretical lambda(max) with the experimental values reported in literature for both compounds. PMID- 19548757 TI - Correlation effects on the dynamics of bipolarons in nondegenerate conjugated polymers. AB - By employing an adaptive time-dependent density-matrix-renormalization-group method, we investigate the dynamics of a charged bipolaron in the presence of both electron-phonon and electron-electron interactions. We use a Su-Schrieffer Heeger model modified to include electron-electron interactions via a Hubbard Hamiltonian, a Brazovskii-Kirova symmetry-breaking term, and an external electric field. Our results show that the velocity of the bipolaron increases first and then decreases with the increasing of the on-site Coulomb interaction, U. Furthermore, the dependence of the bipolaron velocity, bipolaron effective mass, and bipolaron stability on the lattice structures is discussed. PMID- 19548758 TI - Temperature effects on quasi-isolated conjugated polymers as revealed by temperature-dependent optical spectra of 16-mer oligothiophene diluted in a sold matrix. AB - Temperature dependences (4-300 K) of photoluminescence (PL) and absorption spectra of 16-mer oligothiophene (16 T) extremely diluted in polypropylene (PP) have been investigated in order to clarify temperature effects on quasi-isolated conjugated polymers. The PL and absorption spectra are found to blueshift with increasing temperature. The reason for the blueshift is discussed by comparing models based on the refractive index of the solvent (PP) and on the thermal conformational change of 16 T. The blueshift is concluded to result from the thermal conformational change. Time-resolved PL spectra show a redshift of PL band following photoexcitation (spectral migration). The amount of the migration is shown to increase with increasing temperature. The increased migration is concluded to be due to the thermal conformational change. The temperature dependence of the effective conjugation length (ECL) of 16 T is calculated for the absorption and PL transitions. The calculation suggests that ECL is reduced at room temperature to two-thirds of the intrinsic chain length. The activation energy of the conformational change is estimated to be 22.4 meV from the temperature dependence of ECL. We demonstrate that the steady-state PL spectra are well reproduced by simple Franck-Condon analyses using a single Huang-Ryes factor over a wide temperature range. The analyses reveal features of temperature dependence in important spectral parameters such as the Stokes shift, linewidth, and Huang-Ryes factor. PMID- 19548759 TI - Glassy dynamics of liquid crystalline 4(')-n-pentyl-4-cyanobiphenyl in the isotropic and supercooled nematic phases. AB - The temperature evolution of dielectric relaxation times (tau(T)) in the isotropic (I) and supercooled nematic (N) phases of 4(')-n-pentyl-4-cyanobiphenyl is discussed. This compound may be considered as the classical model-material for the physics of liquid crystals and recently also for the glass transition physics. Experimental results extends from approximately T(I-N)+100 K down to T(g) approximately T(I-N)-100 K, where T(I-N) denotes the I-N phase transition temperature. The supporting derivative-based analysis enabled the distortion sensitive tests of the validity of tau(T) parametrization using the Vogel-Fulcher Tammann, Avramov model, and "critical-like" equations. Only the latter yielded the possibility of portraying tau(T) data using a single equation for the isotropic phase within the mode coupling theory and for the supercooled nematic phase within the dynamical scaling model. Issues related to the fragility of the system are also discussed. The existence of the hidden N-SmA transition, damped due to the earlier vitrification at the glass temperature (T(g)), is speculatively suggested. For the isotropic phase the supplementary discussion of dynamics in 4(')-n-butyl-4-cyanobiphenyl is presented. PMID- 19548760 TI - Generalized van der Waals theory for the twist elastic modulus and helical pitch of cholesterics. AB - We present a generalized van der Waals theory for a lyotropic cholesteric system of chiral spherocylinders based on the classical Onsager theory for hard anisometric bodies. The rods consist of a hard spherocylindrical backbone surrounded with a square-well potential to account for attractive (or soft repulsive) interactions. Long-ranged chiral interactions are described by means of a simple pseudoscalar potential which is appropriate for weak chiral forces of a predominant electrostatic origin. Based on the formalism proposed by Straley [Phys. Rev. A 14, 1835 (1976)], we derive explicit algebraic expressions for the twist elastic modulus and the cholesteric pitch for rods as a function of density and temperature. The pitch varies nonmonotonically with density, with a sharp decrease at low packing fractions and a marked increase at higher packing fractions. A similar trend is found for the temperature dependence. The unwinding of the helical pitch at high densities (or low temperatures) originates from a strong enhancement of the local nematic order and the corresponding increase in the twist elastic resistance associated with near-parallel local rod configurations. This contrasts with the commonly held view that the increase in pitch with decreasing temperature as often observed in cholesterics is due to layer formation resulting from presmectic fluctuations. The increase in pitch with increasing temperature is consistent with an entropic unwinding as the chiral interaction becomes less significant than the thermal energy. The variation of the pitch with density, temperature, and contour length is in qualitative agreement with recent experimental results on colloidal fd rods. PMID- 19548761 TI - Correlation of mass fractal dimension and asymmetry. AB - Small-angle X-ray experiments reveal the structure and structural changes of silica embedded in a rubbery host matrix due to the deformation of the surrounding elastomer. The experiments prove that the silica is initially isotropic and becomes anisotropic due to the deformation of the matrix. Increasing the elongation ratio of the polymer results in a larger mass fractal dimension of the silica clusters. The growing mass fractal dimension can be explained simply by a rearrangement of the primary particles within the clusters. However, for the first time, mathematical reasons are presented which clearly demonstrate that self-affine clusters have to be used instead of self-similar ones in order to describe the experiments correctly. PMID- 19548762 TI - The impact of hydration water on the dynamics of side chains of hydrophobic peptides: from dry powder to highly concentrated solutions. AB - Elastic and quasielastic neutron scattering experiments are used to investigate the dynamics of side chains in proteins, using hydrophobic peptides, from dry and hydrated powders up to solutions, as models. The changes of the internal dynamics of a prototypical hydrophobic amino acid, N-acetyl-leucine-methylamide, and alanine amino acids are investigated as a function of water/peptide molecular ratio. While previous results have shown that, in concentrated solution, when the hydrophobic side chains are hydrated by a single hydration water layer, the only allowed motions are confined and can be attributed to librational/rotational movements associated with the methyl groups. In the present work we observe a dynamical evolution from dry to highly hydrated powder. We also observe rotational and diffusive motions and a dynamical transition at approximately 250 K for long side chain peptides while for peptides with short side chains, there is no dynamical transition but only rotational motions. With a local measurement of the influence of hydration water dynamics on the amino acid side chains dynamics, we provide unique experimental evidence that the structural and dynamical properties of interfacial water strongly influence the side chain dynamics and the activation of diffusive motions. We also emphasize that the side chain length has a role on the onset of dynamical transition. PMID- 19548763 TI - Multipili attachment of bacteria with helixlike pili exposed to stress. AB - A number of biomechanical properties of various types of pili expressed by Escherichia coli, predominantly their force-versus-elongation behavior, have previously been assessed in detail on a single pilus level. In vivo, however, bacteria bind in general to host cells by a multitude of pili, which presumably provides them with adhesion properties that differs from those of single pili. Based upon the previously assessed biomechanical properties of individual pili, this work presents a theoretical analysis of the adhesion properties of multipili attaching bacteria expressing helixlike pili exposed to an external force. Expressions for the adhesion lifetime of dual- and multipili-attaching bacteria are derived and their validity is verified by Monte Carlo simulations. It is demonstrated that the adhesion lifetime of a multipili-binding bacterium depends to a large degree on the cooperativity of the attaching pili, which, in turn, depends strongly on their internal biomechanical properties, in particular their helixlike structure and its ability to elongate, which, in turn, depends on the intrinsic properties of the bonds, e.g., their lengths and activation energies. It is shown, for example, that a decrease in the length of a layer-to-layer bond in the rod of P pili, expressed by E. coli, by 50% leads to a decrease in the adhesion lifetime of a bacterium attaching by ten pili and exposed to a force of 500 pN by three orders of magnitude. The results indicate moreover that the intrinsic properties of the rod for this particular type of pili are optimized for multipili attachment under a broad range of external forces and presumably also to its in vivo environment. For example, P pili seems to be optimized to withstand a force exposure during approximately 3 s, which correspond to the time it takes for a bolus to pass a bacterium attached to the ureteral wall. Even though the results presented in this work apply quantitatively to one type of pilus, they are assumed to apply qualitatively to all helixlike pili systems expressing slip bonds. PMID- 19548764 TI - Robustness of integrin signaling network. AB - Integrin signaling network is responsible for regulating a wide variety of fundamental biological processes ranging from cell survival to cell death. While individual components of the network have been studied through experimental and computational methods, the network robustness and the flow of information through the network have not been characterized in a quantitative framework. Using a probability based model implemented through GRID computing, we approach the reduced signaling network and show that the network is highly robust and the final stable steady state is independent of the initial configurations. However, the path from the initial and the final state is intrinsically dependent on the state of the input nodes. Our results demonstrate a rugged funnel-like landscape for the signaling network where the final state is unique, but the paths are dependent on initial conditions. PMID- 19548765 TI - Dependence on temperature and guanine-cytosine content of bubble length distributions in DNA. AB - We present numerical results on the temperature dependence of the distribution of bubble lengths in DNA segments of various guanine-cytosine (GC) concentrations. Base-pair openings are described by the Peyrard-Bishop-Dauxois model and the corresponding thermal equilibrium distributions of bubbles are obtained through Monte Carlo calculations for bubble sizes up to the order of a hundred base pairs. The dependence of the parameters of bubble length distribution on temperature and the GC content is investigated. We provide simple expressions which approximately describe these relations. The variation of the average bubble length is also presented. We find a temperature dependence of the exponent c that appears in the distribution of bubble lengths. If an analogous dependence exists in the loop entropy exponent of real DNA, it may be relevant to understand overstretching in force-extension experiments. PMID- 19548766 TI - Kinetics of proton pumping in cytochrome c oxidase. AB - We propose a simple model of cytochrome c oxidase, including four redox centers and four protonable sites, to study the time evolution of electrostatically coupled electron and proton transfers initiated by the injection of a single electron into the enzyme. We derive a system of master equations for electron and proton state probabilities and show that an efficient pumping of protons across the membrane can be obtained for a reasonable set of parameters. All four experimentally observed kinetic phases appear naturally from our model. We also calculate the dependence of the pumping efficiency on the transmembrane voltage at different temperatures and discuss a possible mechanism of the redox-driven proton translocation. PMID- 19548768 TI - A new water anomaly: the temperature dependence of the proton mean kinetic energy. AB - In a recent article [Pietropaolo et al., Phys. Rev. Lett.100, 127802 (2008)] we discussed the results of a deep inelastic neutron scattering experiment aimed at the measure of mean kinetic energy, , and momentum distribution of protons in supercooled water. The observed excess of proton mean kinetic energy, with respect to theoretical predictions and measurements in water stable liquid and solid phases, suggested a possible link between the anomalous temperature dependence of water density and the temperature dependence of . Nevertheless, the limited number of data did not allow a more quantitative description. Here we report on new measurements, above and below the temperature of maximum density and in the supercooled phase. These show two maxima: The first one, in the supercooled phase in the range of 269-272 K, and a second one at 277 K, the temperature of the maximum density of water. PMID- 19548767 TI - Generic coarse-grained model for protein folding and aggregation. AB - A generic coarse-grained (CG) protein model is presented. The intermediate level of resolution (four beads per amino acid, implicit solvent) allows for accurate sampling of local conformations. It relies on simple interactions that emphasize structure, such as hydrogen bonds and hydrophobicity. Realistic alpha/beta content is achieved by including an effective nearest-neighbor dipolar interaction. Parameters are tuned to reproduce both local conformations and tertiary structures. The thermodynamics and kinetics of a three-helix bundle are studied. We check that the CG model is able to fold proteins with tertiary structures and amino acid sequences different from the one used for parameter tuning. By studying both helical and extended conformations we make sure the force field is not biased toward any particular secondary structure. The accuracy involved in folding not only the test protein but also other ones show strong evidence for amino acid cooperativity embedded in the model. Without any further adjustments or bias a realistic oligopeptide aggregation scenario is observed. PMID- 19548771 TI - Synthesis, characterization, antimicrobial activity and structure-activity relationships of new aryldisulfonamides. AB - A series of aromatic disulfonamide (1-8) derivatives and 4-methylbenzenesulfonyl hydrazide (9) were synthesized and characterized. All compounds were evaluated in vitro for their antimicrobial activity against Staphylococcus aureus ATCC 25953, Bacillus cereus ATCC 6633, Bacillus magaterium RSKK 5117, Escherichia coli ATCC 11230, Salmonella enterititis ATCC 13076 by microdilution and disc diffusion methods. Antimicrobial activity of the aromatic disulfonamides decreased as the length of the carbon chain increased. An analysis of the structure- activity relationship (SAR) along with computational studies showed that the most active compound (9) possessed low lipophilicity (AlogP=0.59) and high solubility (logS = -1.33). PMID- 19548770 TI - Induced multilineage differentiation of chicken embryonic germ cells via embryoid body formation. AB - Although the pluripotent and proliferative capacity of embryonic germ (EG) cells is thought to be equivalent to that of embryonic stem (ES) cells, there has been far less attention focused on the potential use of EG cells for applications in developing novel strategies of tissue transplantation in the treatment of degenerative diseases. In this study, EG cells were derived from primordial germ cells (PGCs) of genital ridges of 4-day-old chicken embryos. These cells satisfied the criteria previously used for defining chicken EG cells by using the expression of markers characteristic to ES cells. When injected subcutaneously, chicken EG cells could form teratomas that enable differentiation into a wide range of tissue types of all three primary cell lineages including neural cells, cartilage, forming bone, adipocytes, blood vessels, smooth muscle, and secretory epithelia in the recipients. Furthermore, cells in embryoid bodies (EBs) expressed lineage-specific markers of three germ layers and could be induced to differentiate into more advanced stages of various committed cell types, including dopamine and cholinergic neurons, astrocytes, oligodendrocytes, adipocytes, and hepatocytes, which were demonstrated by immunocytochemical staining or RT-PCR analysis. These findings support the multilineage differentiation capability of chicken pluripotent EG cells, thus confirming the presumption that chicken embryos may be used as a potential model for better understanding the mechanisms of tissue-specific differentiation and regeneration that will help to devise strategies based on the transplantation of stem cell derived tissues for restoring function to damaged or diseased tissues. PMID- 19548772 TI - Bortezomib-induced perivascular dermatitis in a patient with multiple myeloma. AB - Bortezomib is a potent, selective proteasome inhibitor. It is usually well tolerated, and bortezomib-induced skin lesions are less well known. A 59-year-old man with multiple myeloma underwent autologous hematopoetic stem cell transplantation (AHSCT). Administration of bortezomib was started after AHSCT as his disease progressed despite thalidomide maintenance. He developed a rash afer the third cycle of bortezomib. A biopsy showed superficial and deep perivascular dermatitis. The rash resolved within 4 days; complete response of myeloma was obtained after 8 cycles of bortezomib. The benign clinical course did not require interruption of the treatment, which in turn resulted in complete remission of myeloma. PMID- 19548773 TI - Isoprostanes 8-iPF2alpha-III: risk markers of premature rupture of fetal membranes? AB - AIMS: Isoprostanes may serve as sensitive and specific markers of in vivo oxidative stress intensity. We wanted to determine, whether or not isoprostane concentration may be considered as a risk marker of premature rupture of fetal membranes (PROM). METHODS: On the basis of the presence of PROM and gestational maturity, a total of 128 patients were divided into: (1) preterm PROM (pPROM) group; (2) PROM at term group; (3) control preterm (C1) group and (4) control at term (C2) group. The concentrations of 8-iPF(2alpha)-III were determined using the enzyme-linked immunosorbent assay method. RESULTS: The mean free isoprostane concentrations, examined in amniotic fluid and maternal plasma in the PROM at term patients were significantly higher than in C2 individuals (p < 0.01). The mean concentrations of free 8-iPF(2alpha)-III measured in blood plasma from women in the C1 group were significantly lower than in patients from the pPROM, PROM at term and C2 groups (p < 0.001, p < 0.00001 and p < 0.00001, respectively). CONCLUSION: The measurement of free isoprostane concentration in maternal plasma and amniotic fluid may be considered as a laboratory marker of a PROM-risk pregnancy. PMID- 19548774 TI - Serum Nogo-A levels are not elevated in amyotrophic lateral sclerosis patients. AB - Improved biomarkers would facilitate the diagnosis and treatment of amyotrophic lateral sclerosis (ALS). Muscle content of the neuritic outgrowth inhibitor Nogo A is increased in patients with ALS and other denervating conditions. Seeking a less invasive diagnostic method, we sought to determine whether or not Nogo increases in the serum of ALS patients. We developed a dissociation-enhanced lanthanide fluorescent immunoassay (DELFIA) protocol to screen serum samples from 172 ALS patients and 172 healthy controls for Nogo-A immunoreactivity. Unexpectedly, there was a trend toward decreased levels of serum Nogo-A in ALS. Mean serum Nogo-A level in ALS patients was 0.71 nM (95% confidence interval (CI) 0.42-1.00), as opposed to 1.15 nM (95% CI 0.72-1.59) in healthy controls. A significantly larger percentage of healthy control sera (11.0% vs 4.7%) displayed markedly elevated levels of Nogo-A. Additional study is required to determine the factors that lead to elevated Nogo-A levels in a subset of both ALS patients and healthy controls. PMID- 19548775 TI - Blood gas partition coefficient and pulmonary extraction ratio for propofol in goats and pigs. AB - The interpretation of continuously measured propofol concentration in respiratory gas demands knowledge about the blood gas partition coefficient and pulmonary extraction ratio for propofol. In the present investigation we compared both variables for propofol between goats and pigs during a propofol anaesthesia. In ten goats and ten pigs, expired alveolar gas and arterial and mixed venous blood samples were simultaneously drawn during total intravenous anaesthesia with propofol. The blood gas partition coefficient and pulmonary extraction ratio were calculated for both species. Non-parametric methods were used for statistical inference. The blood gas partition coefficient ranged between 7000 and 646,000 for goats and between 17,000 and 267,000 for pigs. The pulmonary extraction ratio ranged between 32.9% and 98.1% for goats and was higher for pigs, which ranged between -106.0% and 39.0%. The blood gas partition coefficient for propofol exceeded those for other known anaesthetic compounds so that it takes longer to develop a steady-state. The different pulmonary extraction rates in two species suggest that there are different ways to distribute propofol during the lung passage on its way from the blood to breathing gas. This species-specific difference has to be considered for methods using the alveolar gas for monitoring the propofol concentration in plasma. PMID- 19548776 TI - Evidence supporting a role for endoplasmic reticulum stress in the development of atherosclerosis in a hyperglycaemic mouse model. AB - We previously observed a correlation between elevated levels of vascular endoplasmic reticulum (ER) stress and accelerated atherosclerotic plaque development in chronically hyperglycemic apolipoprotein-deficient (ApoE(-/-)) mice. We hypothesize that ER stress plays a causative role in diabetic atherogenesis. Here we examine the temporal relation between the onset of hyperglycemia, glucosamine accumulation in the vessel wall, ER stress, and the development of atherosclerosis. We demonstrate, by using streptozotocin-induced hyperglycemic ApoE(-/-) mice, that conditions of hyperglycemia increase intracellular glucosamine levels and endothelial ER stress levels in the endothelium before the onset of atherosclerosis. At 15 weeks of age, hyperglycemic mice have significantly larger atherosclerotic lesions (0.120 +/- 0.023 vs. 0.065 +/- 0.021 mm2; p = 0.001) relative to normoglycemic mice. Significantly, hyperglycemia-associated accelerated atherosclerosis is observed before the onset of dyslipidemias, suggesting that leveled glucose is sufficient to promote atherogenesis independently. Diagnostic markers of elevated ER-stress levels are increased in macrophage-derived foam cells in early and advanced atherosclerotic lesions. Dietary supplementation with valproate, a small branched chain fatty acid that interferes with ER-stress signaling, significantly attenuates accelerated atherogenesis in this model. Together, these data are consistent with a causative role for hyperglycemia-associated ER stress in the development and progression of diabetic atherosclerosis. PMID- 19548777 TI - Inhibition of protein tyrosine phosphatase 1B by lupeol and lupenone isolated from Sorbus commixta. AB - Protein tyrosine phosphatase 1B (PTP1B) appears to be an attractive target for the development of new drugs for type 2 diabetes and obesity. In our preliminary test, a MeOH extract of the stem barks of Sorbus commixta Hedl. (Rosaceae) showed strong PTP1B inhibitory activity. Bioassay-guided fractionation of the MeOH extract resulted in the isolation of two lupane-type triterpenes, lupenone (1) and lupeol (2). Compounds 1 and 2 inhibited PTP1B with IC(50) values of 13.7 +/- 2.1 and 5.6 +/- 0.9 microM, respectively. Kinetic studies revealed that both the compounds 1 and 2 are non-competitive inhibitors of PTP1B that decrease V(max) values with no effect on K(m) values. PMID- 19548780 TI - Antioxidant phenylpropanoid glycosides from Buddleja davidii. AB - Phytochemical investigations on the n-BuOH-soluble fraction of the whole plant of Buddleja davidii led to the isolation of the phenylpropanoid glycosides 1-10. Their structures were determined by 1D and 2D NMR spectroscopic techniques. All the compounds showed potent antioxidative activity in three different tests, with IC(50) values in the range 4.15-9.47 microM in the hydroxyl radical ( OH) inhibitory activity test, 40.32-81.15 microM in the total ROS (reactive oxygen species) inhibitory activity test, and 2.26-7.79 microM in the peroxynitrite (ONOO(-)) scavenging activity test. Calceolarioside A (1) displayed the strongest scavenging potential with IC(50) values of (4.15 +/- 0.07, 40.32 +/- 0.09, 2.26 +/- 0.03 microM) for OH, total ROS and scavenging of ONOO(-), respectively. PMID- 19548781 TI - Modeling of the growth hormone secretagogue receptor antagonistic activity using chemometric tools. AB - A quantitative structure-activity relationship (QSAR) study has been carried out on growth hormone secretagogue receptor antagonistic activity of the derivatives of 2,4-diaminopyrimidine. To obtain significant QSARs, the approaches involving the non-parametric such as Fujita-Ban, and the parametric based on physicochemical and DRAGON descriptors in Hansch type of analysis have been employed. The Fujita-Ban approach, however, was constrained to 18 compounds only due to a limited number of substituents appeared at varying positions. The derived contributions of different substituents and the parent moiety were used to identify the potential congeners. The physicochemical model of Hansch was subsequently used to interpret the type of interactions involved between the receptor sites and varying positions of these compounds. The study, employing DRAGON descriptors in Hansch approach was also carried out on this data set to discuss the prevailing interactions in terms of topological descriptors. The derived highest significant model was discussed to delineate the type of interactions involved and suggestions have been made for different alterations to lead to further potential compounds of the series. PMID- 19548782 TI - Effect of some analgesics on paraoxonase-1 purified from human serum. AB - The in vitro effects of the analgesic drugs, lornoxicam, indomethacin, tenoxicam, diclofenac sodium, ketoprofen and lincomycine, on the activity of purified human serum paraoxonase (hPON1) (EC 3.1.8.1.) were evaluated. hPON1 was purified from human serum with a final specific activity of 3840 U mg(-1) and a purity of 25.3 % using simple chromatographic methods, including DEAE-Sephadex anion exchange and Sepharose 4B-L-tyrozine-1-napthylamine hydrophobic interaction chromatography. SDS-polyacrylamide gel electrophoresis indicated a single protein band corresponding to hPON1. The six analgesics dose-dependently decreased in vitro hPON1 activity, with IC(50) values for lornoxicam, indomethacin, tenoxicam, diclofenac sodium, ketoprofen and lincomycine of 0.136, 0.195, 0.340, 1.639, 6.23 and 9.638 mM, respectively. K(i) constants were 0.009, 0.097, 0.306, 0.805, 13.010 and 11.116 mM, respectively. Analgesics showed different inhibition mechanisms: lornoxicam, diclofenac sodium and lincomycine were uncompetitive, indomethacin and tenoxicam were competitive, ketoprofen was noncompetitive. According to the results, inhibition potency was lornoxicam>indomethacin>tenoxicam> diclofenac sodium>ketoprofen> lincomycine. PMID- 19548783 TI - Co-creating meaningful structures within long-term psychotherapy group culture. AB - Meaningful group structures are co-created within the long-term outpatient psychotherapy group through a hermeneutical interaction between structure and immediate experience of structure by individuals embedded in personal and collective contexts. Co-created meanings expand original group- and self understandings and further evolve structures that are stable yet do not exist independently of the narratives and affects of the members who interact with them. Group structures do not reduce, expand, or dissolve but change in connection to the experiences and meaning attributions within the group. This intersubjective process mediates the emphasis within group theory on leader responsibility for culture building that risks overpromoting certain psychotherapeutic cultural intentions over others. Three examples of intersubjective hermeneutical interaction within long-term psychotherapy groups lend insight into global, cultural, and societal groups. PMID- 19548784 TI - The conversation group: using group psychoanalytic techniques to resolve resistances of recently immigrated Chinese students to learning English in a high school setting. AB - Does group psychoanalytic theory and technique have an application in an ordinary high school classroom? In this article, the writer describes a research project in which she attempts to answer this question by applying the techniques with a group of recently immigrated Chinese students who wished to improve their spoken English. PMID- 19548785 TI - Cognitive behavior therapy for early psychosis: a comprehensive review of individual vs. group treatment studies. AB - Several recent studies of individually administered cognitive behavior therapy (CBT) for early psychosis have reported only modest treatment benefits. The purpose of the current study was to review the literature to determine how outcomes of group CBT differ from outcomes of individually administered CBT among early cases. Our findings suggest that group CBT for early psychosis may be a more effective modality for this group of patients. We speculate that patients' uncertainty about illness in general may impair the effectiveness of individually administered CBT for early cases and that group CBT may be more effective for these young patients by better addressing those factors with the aid of peer-to peer interactions, identification, and modeling. PMID- 19548786 TI - A psychoeducational group for adolescent girls to facilitate egalitarian, non abusive relationships. AB - Violence in Israeli society has in recent years become more prevalent. Our Social and Welfare Services have been receiving more reports of physical, psychological, and sexual abuse within families and within Israeli society at large. In an attempt to address this problem, the authors selected eight girls, 14 to 15 years of age, to participate in a group to explore egalitarian relationships, with the goal of reducing the likelihood that members will enter a violent relationship. The group was part of a preventive intervention program for girls who were identified as being at high risk. The participants met for a 12-session psychoeducational therapy group to discuss a range of different topics associated with egalitarian relationships and violence, from both theoretical and practical perspectives. After having completed the program, participants generally reported a high level of satisfaction. The girls mostly responded that participating in the group greatly enhanced their knowledge about egalitarian, non-abusive relationships, as well as their personal awareness of the implications of violent behavior. PMID- 19548787 TI - A model parent group for enhancing aggressive children's social competence in Taiwan. AB - This paper presents a semi-structured psychoeducational model of group work for parents of aggressive children based on concepts of co-parenting and bidirectionality. The group was developed for enhancing five Taiwanese aggressive children's social competence by promoting positive interactions within family. Topics covered in the group included identifying parenting styles, forming parental alliances, fostering parent-child mutual initiations/mutual compliances, establishing parent-child co-regulation, and responding to aggressive children's negative emotions. Pre- and post-group comparisons suggested the effectiveness of the group model. PMID- 19548788 TI - Affect education and the development of the interpersonal ego in modern group psychoanalysis. PMID- 19548790 TI - Multifamily group treatment for schizophrenia. PMID- 19548793 TI - Pair wise binding affinity: 3D QSAR studies on a set of triazolo [1, 5-a] quinoxalines as antagonists of AMPA and KA receptors. AB - The glutamate receptor system is implicated in the development and maintenance of epileptic seizures and it has been reported that compounds showing high affinity for both AMPA and KA binding sites are more potent anticonvulsants than compounds having selective affinity toward AMPA or KA receptor. These outcomes make such inhibitors future potential antiepileptic drugs. So, the pair wise binding affinity for AMPA and KA receptors inhibition was proposed by using the addition between biological activities of ligands. This approach for evaluation of pair wise binding affinity was exemplified using set of triazolo [1,5-a] quinoxaline for AMPA and KA receptors. The biological activity towards AMPA and KA receptors (expressed as -log IC(5O)) was taken as a dependent variable for building CoMFA and CoMSIA models. The resulting models show the ways of increasing binding affinity to both AMPA and KA receptors as potential target for epilepsy. The statistically significant results show that pair wise CoMFA and CoMSIA models are better then individual models. The resulting cross-validated r(2)(CV) value 0.806 for CoMFA is greater then 0.780 for CoMSIA pair wise model. The non-cross validated run giving a coefficient of determination r(2) value of 0.946 and 0.908 for CoMFA and CoMSIA respectively, provided a good correlation between the observed and computed affinities of the compounds. PMID- 19548794 TI - Update on biochemical properties of recombinant Pseudomonas diminuta phosphotriesterase. AB - Phosphotriesterase from Pseudomonas diminuta (PTE; EC 3.1.8.1) hydrolyzes organophosphate insecticides and chemical warfare agents. The two zinc cations in the active center can be substituted. Co(2+)-containing PTE is the most efficient but least stable isoform. Gel filtration showed that PTE is monomeric at the submicromolar concentrations used in kinetic assays. The analysis of the recombinant enzyme by X-ray fluorescence spectrometry and CCT-ICP-MS, confirms that recombinant Zn-PTE contains only Zn(2+) whereas Co-PTE has Zn(2+) and Co(2+) in equimolar amount, with Co(2+) most likely in the reported labile beta-site. We noted that recombinant PTE is unstable at low concentrations and must be stabilized by a protein environment. We tested the effect of excess of various metal cofactors on PTE-catalyzed hydrolysis of paraoxon. We notably observed that ZnCl(2) induces a non-competitive partial inhibition of Zn(2+)- and Co(2+)-PTE at pH 8.5 (apparent Ki=155 microM and 52 microM, respectively). Inhibition results from interactions with colloidal Zn(OH)(2) formed in alkaline buffer that alters the catalytic machinery. NiCl(2) caused a similar effect at higher concentrations (apparent Ki=3 mM). We observed that mutating His123, a surface residue close to an alleged allosteric site, dramatically altered the bacterial expression yield of Co(2+)-PTE, Ki for Zn(OH)(2) inhibition, k(cat) (up to 60 fold) for paraoxon hydrolysis, but not K(M). Issues addressed in this work are important for future biotechnological developments of PTE as a detoxifying enzyme. PMID- 19548795 TI - Computation with spikes in a winner-take-all network. AB - The winner-take-all (WTA) computation in networks of recurrently connected neurons is an important decision element of many models of cortical processing. However, analytical studies of the WTA performance in recurrent networks have generally addressed rate-based models. Very few have addressed networks of spiking neurons, which are relevant for understanding the biological networks themselves and also for the development of neuromorphic electronic neurons that commmunicate by action potential like address-events. Here, we make steps in that direction by using a simplified Markov model of the spiking network to examine analytically the ability of a spike-based WTA network to discriminate the statistics of inputs ranging from stationary regular to nonstationary Poisson events. Our work extends previous theoretical results showing that a WTA recurrent network receiving regular spike inputs can select the correct winner within one interspike interval. We show first for the case of spike rate inputs that input discrimination and the effects of self-excitation and inhibition on this discrimination are consistent with results obtained from the standard rate based WTA models. We also extend this discrimination analysis of spiking WTAs to nonstationary inputs with time-varying spike rates resembling statistics of real world sensory stimuli. We conclude that spiking WTAs are consistent with their continuous counterparts for steady-state inputs, but they also exhibit high discrimination performance with nonstationary inputs. PMID- 19548796 TI - Orientation disparity: a cue for 3D orientation? AB - Orientation disparity, the difference in orientation that results when a texture element on a slanted surface is projected to the two eyes, has been proposed as a binocular cue for 3D orientation. Since orientation disparity is confounded with position disparity, neither behavioral nor neurophysiological experiments have successfully isolated its contribution to slant estimates or established whether the visual system uses it. Using a modified disparity energy model, we simulated a population of binocular visual cortical neurons tuned to orientation disparity and measured the amount of Fisher information contained in the activity patterns. We evaluated the potential contribution of orientation disparity to 3D orientation estimation and delimited the stimulus conditions under which it is a reliable cue. Our results suggest that orientation disparity is an efficient source of information about 3D orientation and that it is plausible that the visual system could have mechanisms that are sensitive to it. Although orientation disparity is neither necessary nor sufficient for estimating slant, it appears that it could be useful when combined with estimates from position disparity gradients and monocular perspective cues. PMID- 19548797 TI - Sequential Monte Carlo point-process estimation of kinematics from neural spiking activity for brain-machine interfaces. AB - Many decoding algorithms for brain machine interfaces' (BMIs) estimate hand movement from binned spike rates, which do not fully exploit the resolution contained in spike timing and may exclude rich neural dynamics from the modeling. More recently, an adaptive filtering method based on a Bayesian approach to reconstruct the neural state from the observed spike times has been proposed. However, it assumes and propagates a gaussian distributed state posterior density, which in general is too restrictive. We have also proposed a sequential Monte Carlo estimation methodology to reconstruct the kinematic states directly from the multichannel spike trains. This letter presents a systematic testing of this algorithm in a simulated neural spike train decoding experiment and then in BMI data. Compared to a point-process adaptive filtering algorithm with a linear observation model and a gaussian approximation (the counterpart for point processes of the Kalman filter), our sequential Monte Carlo estimation methodology exploits a detailed encoding model (tuning function) derived for each neuron from training data. However, this added complexity is translated into higher performance with real data. To deal with the intrinsic spike randomness in online modeling, several synthetic spike trains are generated from the intensity function estimated from the neurons and utilized as extra model inputs in an attempt to decrease the variance in the kinematic predictions. The performance of the sequential Monte Carlo estimation methodology augmented with this synthetic spike input provides improved reconstruction, which raises interesting questions and helps explain the overall modeling requirements better. PMID- 19548798 TI - Evaluation of trajectory planning models for arm-reaching movements based on energy cost. AB - Computational studies have suggested that many characteristics of reaching trajectories in a horizontal plane can be effectively predicted by certain models, including, the minimum end point variance model and minimum torque change model. It has also been reported that these characteristics appear to differ from those obtained by the minimum energy cost model that has been reported to explain the characteristics of locomotor patterns. Do these results imply that the human nervous system uses different strategies to resolve the redundancy problem for different tasks? In order to reexamine the optimality of reaching trajectories from a viewpoint of energy cost, we considered the corrective submovements to compensate for positional error due to signal-dependent noise in motor commands and computed the expected value of the total energy costs required to reach a target by repetition of submovements planned by each of the following models: the minimum energy cost model, minimum end point variance model, and minimum torque change model. The results revealed that when the noise is large, the total energy cost required by the minimum end point variance model and the minimum torque change model can be lower than that required by the minimum energy cost model which assumes minimizing energy cost under noise-free condition. This result indicates that the minimization of the expected value of the energy cost would be an important factor in determining the reaching trajectories. PMID- 19548799 TI - A model for learning topographically organized parts-based representations of objects in visual cortex: topographic nonnegative matrix factorization. AB - Object representation in the inferior temporal cortex (IT), an area of visual cortex critical for object recognition in the primate, exhibits two prominent properties: (1) objects are represented by the combined activity of columnar clusters of neurons, with each cluster representing component features or parts of objects, and (2) closely related features are continuously represented along the tangential direction of individual columnar clusters. Here we propose a learning model that reflects these properties of parts-based representation and topographic organization in a unified framework. This model is based on a nonnegative matrix factorization (NMF) basis decomposition method. NMF alone provides a parts-based representation where nonnegative inputs are approximated by additive combinations of nonnegative basis functions. Our proposed model of topographic NMF (TNMF) incorporates neighborhood connections between NMF basis functions arranged on a topographic map and attains the topographic property without losing the parts-based property of the NMF. The TNMF represents an input by multiple activity peaks to describe diverse information, whereas conventional topographic models, such as the self-organizing map (SOM), represent an input by a single activity peak in a topographic map. We demonstrate the parts-based and topographic properties of the TNMF by constructing a hierarchical model for object recognition where the TNMF is at the top tier for learning high-level object features. The TNMF showed better generalization performance over NMF for a data set of continuous view change of an image and more robustly preserving the continuity of the view change in its object representation. Comparison of the outputs of our model with actual neural responses recorded in the IT indicates that the TNMF reconstructs the neuronal responses better than the SOM, giving plausibility to the parts-based learning of the model. PMID- 19548800 TI - Constraint on the number of synaptic inputs to a visual cortical neuron controls receptive field formation. AB - To date, Hebbian learning combined with some form of constraint on synaptic inputs has been demonstrated to describe well the development of neural networks. The previous models revealed mathematically the importance of synaptic constraints to reproduce orientation selectivity in the visual cortical neurons, but biological mechanisms underlying such constraints remain unclear. In this study, we addressed this issue by formulating a synaptic constraint based on activity-dependent mechanisms of synaptic changes. Particularly, considering metabotropic glutamate receptor-mediated long-term depression, we derived synaptic constraint that suppresses the number of inputs from individual presynaptic neurons. We performed computer simulations of the activity-dependent self-organization of geniculocortical inputs with the synaptic constraint and examined the formation of receptive fields (RFs) of model visual cortical neurons. When we changed the magnitude of the synaptic constraint, we found the emergence of distinct RF structures such as concentric RFs, simple-cell-like RFs, and double-oriented RFs and also a gradual transition between spatiotemporal separable and inseparable RFs. Thus, the model based on the synaptic constraint derived from biological consideration can account systematically for the repertoire of RF structures observed in the primary visual cortices of different species for the first time. PMID- 19548801 TI - Stochastic properties of coincidence-detector neural cells. AB - Neural information is characterized by sets of spiking events that travel within the brain through neuron junctions that receive, transmit, and process streams of spikes. Coincidence detection is one of the ways to describe the functionality of a single neural cell. This letter presents an analytical derivation of the output stochastic behavior of a coincidence detector (CD) cell whose stochastic inputs behave as a nonhomogeneous Poisson process (NHPP) with both excitatory and inhibitory inputs. The derivation, which is based on an efficient breakdown of the cell into basic functional elements, results in an output process whose behavior can be approximated as an NHPP as long as the coincidence interval is much smaller than the refractory period of the cell's inputs. Intuitively, the approximation is valid as long as the processing rate is much faster than the incoming information rate. This type of modeling is a simplified but very useful description of neurons since it enables analytical derivations. The statistical properties of single CD cell's output make it possible to integrate and analyze complex neural cells in a feedforward network using the methodology presented here. Accordingly, basic biological characteristics of neural activity are demonstrated, such as a decrease in the spontaneous rate at higher brain levels and improved signal-to-noise ratio for harmonic input signals. PMID- 19548802 TI - Automatic spike sorting using tuning information. AB - Current spike sorting methods focus on clustering neurons' characteristic spike waveforms. The resulting spike-sorted data are typically used to estimate how covariates of interest modulate the firing rates of neurons. However, when these covariates do modulate the firing rates, they provide information about spikes' identities, which thus far have been ignored for the purpose of spike sorting. This letter describes a novel approach to spike sorting, which incorporates both waveform information and tuning information obtained from the modulation of firing rates. Because it efficiently uses all the available information, this spike sorter yields lower spike misclassification rates than traditional automatic spike sorters. This theoretical result is verified empirically on several examples. The proposed method does not require additional assumptions; only its implementation is different. It essentially consists of performing spike sorting and tuning estimation simultaneously rather than sequentially, as is currently done. We used an expectation-maximization maximum likelihood algorithm to implement the new spike sorter. We present the general form of this algorithm and provide a detailed implementable version under the assumptions that neurons are independent and spike according to Poisson processes. Finally, we uncover a systematic flaw of spike sorting based on waveform information only. PMID- 19548804 TI - Receptive field self-organization in a model of the fine structure in v1 cortical columns. AB - We study a dynamical model of processing and learning in the visual cortex, which reflects the anatomy of V1 cortical columns and properties of their neuronal receptive fields. Based on recent results on the fine-scale structure of columns in V1, we model the activity dynamics in subpopulations of excitatory neurons and their interaction with systems of inhibitory neurons. We find that a dynamical model based on these aspects of columnar anatomy can give rise to specific types of computations that result in self-organization of afferents to the column. For a given type of input, self-organization reliably extracts the basic input components represented by neuronal receptive fields. Self-organization is very noise tolerant and can robustly be applied to different types of input. To quantitatively analyze the system's component extraction capabilities, we use two standard benchmarks: the bars test and natural images. In the bars test, the system shows the highest noise robustness reported so far. If natural image patches are used as input, self-organization results in Gabor-like receptive fields. In quantitative comparison with in vivo measurements, we find that the obtained receptive fields capture statistical properties of V1 simple cells that algorithms such as independent component analysis or sparse coding do not reproduce. PMID- 19548803 TI - Sequential effects in two-choice reaction time tasks: decomposition and synthesis of mechanisms. AB - Performance on serial tasks is influenced by first- and higher-order sequential effects, respectively, due to the immediately previous and earlier trials. As response-to-stimulus interval (RSI) increases, the pattern of reaction times transits from a benefit-only mode, traditionally ascribed to automatic facilitation (AF), to a cost-benefit mode, due to strategic expectancy (SE). To illuminate the sources of such effects, we develop a connectionist network of two mutually inhibiting neural decision units subject to feedback from previous trials. A study of separate biasing mechanisms shows that residual decision unit activity can lead to only first-order AF, but higher-order AF can result from strategic priming mediated by conflict monitoring, which we instantiate in two distinct versions. A further mechanism mediates expectation-related biases that grow during RSI toward saturation levels determined by weighted repetition (or alternation) sequence lengths. Equipped with these mechanisms, the network, consistent with known neurophysiology, accounts for several sets of behavioral data over a wide range of RSIs. The results also suggest that practice speeds up all the mechanisms rather than adjusting their relative strengths. PMID- 19548805 TI - A bound on modeling error in observable operator models and an associated learning algorithm. AB - Observable operator models (OOMs) generalize hidden Markov models (HMMs) and can be represented in a structurally similar matrix formalism. The mathematical theory of OOMs gives rise to a family of constructive, fast, and asymptotically correct learning algorithms, whose statistical efficiency, however, depends crucially on the optimization of two auxiliary transformation matrices. This optimization task is nontrivial; indeed, even formulating computationally accessible optimality criteria is not easy. Here we derive how a bound on the modeling error of an OOM can be expressed in terms of these auxiliary matrices, which in turn yields an optimization procedure for them and finally affords us with a complete learning algorithm: the error-controlling algorithm. Models learned by this algorithm have an assured error bound on their parameters. The performance of this algorithm is illuminated by comparisons with two types of HMMs trained by the expectation-maximization algorithm, with the efficiency sharpening algorithm, another recently found learning algorithm for OOMs, and with predictive state representations (Littman & Sutton, 2001 ) trained by methods representing the state of the art in that field. PMID- 19548806 TI - Belief propagation in networks of spiking neurons. AB - From a theoretical point of view, statistical inference is an attractive model of brain operation. However, it is unclear how to implement these inferential processes in neuronal networks. We offer a solution to this problem by showing in detailed simulations how the belief propagation algorithm on a factor graph can be embedded in a network of spiking neurons. We use pools of spiking neurons as the function nodes of the factor graph. Each pool gathers "messages" in the form of population activities from its input nodes and combines them through its network dynamics. Each of the various output messages to be transmitted over the edges of the graph is computed by a group of readout neurons that feed in their respective destination pools. We use this approach to implement two examples of factor graphs. The first example, drawn from coding theory, models the transmission of signals through an unreliable channel and demonstrates the principles and generality of our network approach. The second, more applied example is of a psychophysical mechanism in which visual cues are used to resolve hypotheses about the interpretation of an object's shape and illumination. These two examples, and also a statistical analysis, demonstrate good agreement between the performance of our networks and the direct numerical evaluation of belief propagation. PMID- 19548807 TI - Rodent-borne diseases and their risks for public health. AB - Rodents are the most abundant and diversified order of living mammals in the world. Already since the Middle Ages we know that they can contribute to human disease, as black rats were associated with distribution of plague. However, also in modern times rodents form a threat for public health. In this review article a large number of pathogens that are directly or indirectly transmitted by rodents are described. Moreover, a simplified rodent disease model is discussed. PMID- 19548814 TI - Functional recovery after the repair of transected cervical roots in the chronic stage of injury. AB - The treatment of root injury is typically performed at the more chronic stages post injury, by which time a substantial number of neurons have died. Therefore, before being applied in the clinical setting, a treatment strategy for these lesions should prove to be as effective in the chronic stages of injury as it is in the acute stage. In this study, we simulated the most severe clinical scenarios to establish an optimal time window for repair at a chronic stage. The sixth to eighth cervical roots on the left side of female SD rats were cut at their junction with the spinal cord. One or three weeks later, the wound was reopened and these roots were repaired with intercostal nerve grafts, with subsequent application of aFGF and fibrin glue. In the control group, the wound was closed after re-exploration without further repair procedures. Sensory and motor functions were measured after the surgery. Spinal cord morphology, neuron survival, and nerve fiber regeneration were traced by CTB-HRP. Results showed that both the sensory and motor functions had significant recovery in the 1-week repair group, but not in the 3-week repair group. By CTB-HRP tracing, we found that the architecture of the spinal cords was relatively preserved in the 1-week repair group, while those of the control group showed significant atrophic change. There were regenerating nerve fibers in the dorsal horn and more motor neuron survival in the 1-week repair group compared to that of the 3-week group. It was concluded that treating transected cervical roots at a chronic stage with microsurgical nerve grafting and application of aFGF and fibrin glue can lead to significant functional recovery, as long as the repair is done before too many neurons die. PMID- 19548817 TI - What is the value proposition? What is the value of telemedicine? PMID- 19548816 TI - No country for old men. PMID- 19548819 TI - Telethinking with Rifat Latifi, M.D. Interview by Vicki Glaser. PMID- 19548820 TI - Telemedicine and autism: researchers and clinicians are just starting to consider telemedicine applications for the diagnosis and treatment of autism. PMID- 19548821 TI - New frontiers: all in the family. PMID- 19548822 TI - Remote assessment of acne: the use of acne grading tools to evaluate digital skin images. AB - Digital imaging of dermatology patients is a novel approach to remote data collection. A number of assessment tools have been developed to grade acne severity and to track clinical progress over time. Although these tools have been validated when used in a face-to-face setting, their efficacy and reliability when used to assess digital images have not been examined. The main purpose of this study was to determine whether specific assessment tools designed to grade acne during face-to-face visits can be applied to the evaluation of digital images. The secondary purpose was to ascertain whether images obtained by subjects are of adequate quality to allow such assessments to be made. Three hundred (300) digital images of patients with mild to moderate facial inflammatory acne from an ongoing randomized-controlled study were included in this analysis. These images were obtained from 20 patients and consisted of sets of 3 images taken over time. Of these images, 120 images were captured by subjects themselves and 180 were taken by study staff. Subjects were asked to retake their photographs if the initial images were deemed of poor quality by study staff. Images were evaluated by two dermatologists-in-training using validated acne assessment measures: Total Inflammatory Lesion Count, Leeds technique, and the Investigator's Global Assessment. Reliability of raters was evaluated using correlation coefficients and kappa statistics. Of the different acne assessment measures tested, the inter-rater reliability was highest for the total inflammatory lesion count (r = 0.871), but low for the Leeds technique (kappa = 0.381) and global assessment (kappa = 0.3119). Raters were able to evaluate over 89% of all images using each type of acne assessment measure despite the fact that images obtained by study staff were of higher quality than those obtained by patients (p < 0.001). Several existing clinical assessment measures can be used to evaluate digital images obtained from subjects with inflammatory acne lesions. The level of inter-rater agreement is highly variable across assessment measures, and we found the Total Inflammatory Lesion Count to be the most reliable. This measure could be used to allow a dermatologist to remotely track a patient's progress over time. PMID- 19548823 TI - Is cardiac consultation with remote-controlled real-time echocardiography a wise use of resources? AB - Northern Sweden is a sparsely populated area with six hospitals and about 50 healthcare centers. The elderly population is a large proportion of the total of population, and the incidence of cardiovascular disease is high. The objective of this research was to analyze the costs and benefits of cardiac consultation in healthcare centers involving long-distance, remote-controlled, real-time echocardiography. The distance diagnostics were developed and tested in two healthcare centers. Experiences of the feasibility of this approach were used as a basis for an economic analysis with regard to heart failure. The societal costs for two different systems were calculated, namely, traditional hospital diagnosis versus distance diagnosis using the new system. The potential prime gainers were the patients. Their traveling time, and thereby their time costs, were significantly reduced. The quality of care may also have been improved. From the health authorities' perspective, the costs of the two systems were approximately equal. Since county council costs are not greatly affected, the large reduction in patient travel time and the improved quality of care ought to be a sufficient incentive for large-scale tests. PMID- 19548824 TI - Tele-transmission of stereoscopic images of the optic nerve head in glaucoma via Internet. AB - The objective was to describe an inexpensive system to visualize stereoscopic photographs of the optic nerve head on computer displays and to transmit such images via the Internet for collaborative research or remote clinical diagnosis in glaucoma. Stereoscopic images of glaucoma patients were digitized and stored in a file format (joint photographic stereoimage [jps]) containing all three dimensional information for both eyes on an Internet Web site (www.trizax.com). The size of jps files was between 0.4 to 1.4 MB (corresponding to a diagonal stereo image size between 900 and 1400 pixels) suitable for Internet protocols. A conventional personal computer system equipped with wireless stereoscopic LCD shutter glasses and a CRT-monitor with high refresh rate (120 Hz) can be used to obtain flicker-free stereo visualization of true-color images with high resolution. Modern thin-film transistor-LCD displays in combination with inexpensive red-cyan goggles achieve stereoscopic visualization with the same resolution but reduced color quality and contrast. The primary aim of our study was met to transmit stereoscopic images via the Internet. Additionally, we found that with both stereoscopic visualization techniques, cup depth, neuroretinal rim shape, and slope of the inner wall of the optic nerve head, can be qualitatively better perceived and interpreted than with monoscopic images. This study demonstrates high-quality and low-cost Internet transmission of stereoscopic images of the optic nerve head from glaucoma patients. The technique allows exchange of stereoscopic images and can be applied to tele-diagnostic and glaucoma research. PMID- 19548825 TI - Usability of robotic platforms for remote surgical teleproctoring. AB - Military field hospitals and rural medical centers may lack surgical subspecialists. Robotic technology can enable proctoring of remotely located general surgeons by subspecialists. Our objective compared three proctoring platforms: (1) 6-degree-of-freedom (DOF) computer input devices controlling a camera and laser pointer mounted on robotic arms, (2) a computer mouse controlling a pan-tilt-zoom (PTZ) camera and robotic laser scanner, and (3) a computer pen/tablet controlling a PTZ-camera and robotic laser scanner. Our hypothesis was that a pen/tablet or mouse platform would be superior to the 6-DOF input device platform. Five surgeons used each platform by simulating the creation of operative incisions. Qualitative (instrument handling, time, motion, spatial awareness) and quantitative performance (accuracy, speed) was assessed on a five-point scale. Each surgeon completed a satisfaction survey. Both mouse and pen/tablet had higher mean performance scores than the 6-DOF-input device in all quantitative (6-DOF = 1.7 +/- 0.8, mouse = 4.3 +/- 0.2, pen = 4.1 +/- 0.6; p < 0.001) and qualitative measures (6-DOF = 1.7 +/- 0.2, mouse = 4.8 +/- 0.0, pen = 4.6 +/- 0.1; p < 0.001). Handling, motion, and instrument awareness were superior with the mouse and pen/tablet versus 6-DOF-input devices (p < 0.0001). Speed and accuracy were also superior using the mouse or pen/tablet versus 6-DOF-input devices (p < 0.0001). Surgeons completed tasks faster using the mouse versus pen/tablet (p = 0.02). Satisfaction surveys revealed a preference for the mouse. This study demonstrates the superiority of a mouse or pen/tablet controlling a PTZ-camera and robotic laser scanner for remote surgical teleproctoring versus 6 DOF-input devices controlling a camera and laser pointer. Either a mouse or pen/tablet platform allows subspecialists to proctor remotely located surgeons. PMID- 19548826 TI - Sharing information and data across heterogeneous e-health systems. AB - Information and data sharing across heterogeneous e-health systems, focusing on the management of patient care, have become the backbone of modern delivery of sustainable telemedicine services. Information and data available to healthcare practitioners in such environments range from patient's medical records, stored in repositories at places where patients have been treated, to a variety of information related to medical research, pharmaceutical products, or information stored within social networks of healthcare interest groups. This study sought to demonstrate two different approaches enabling the sharing of information/data across heterogeneous e-health systems: (1) Context-Aware Data Retrieval Architecture (CADRA), which secures the extraction and presentation of e-health information to users in requested format, and (2) Generic Ontology for Context Aware, Interoperable, and Data Sharing (Go-CID) software applications, which secure semantic interoperation across heterogeneous e-health data sources. Proof of-concept was demonstrated in both cases, CADRA and Go-CID, to achieve understanding and building of knowledge about e-health environments. This study invites practical solutions for interoperable e-health systems. PMID- 19548827 TI - Telemedicine and the mini-mental state examination: assessment from a distance. AB - The objective of this study was to determine the reliability of the Mini-Mental State Examination (MMSE) administration via telehealth with a focus on the auditory and visual test components. Reliability was assessed through use of an in-person collaborator and by assessment of faxed test copies. The MMSE was administered via telehealth with the assistance of a face-to-face collaborator. Patient responses were recorded by both the remote and in-person nurse and compared item by item; total scores for each subject were also compared. Visual items were assessed through a blinded separate scoring of a faxed copy. Percent agreement per item and total score were calculated and correlations between scores were determined by Pearson correlation coefficients. Mean score differences and associated 95% confidence intervals were calculated. Eighty percent of individual items demonstrated remote to in-person agreement of >95% and all items were >85.5% in agreement. Pearson correlation coefficients demonstrated high correlations (>0.86) between 80% of the items examined. Mean differences in scored test items were not significantly different from zero. This study demonstrates the utility of using telehealth for cognitive assessment by MMSE. It supports the use of telehealth to improve healthcare access among patients for whom distance, cost, and mobility are potential barriers to attending face-to-face clinical visits. Continued validation and reliability testing is warranted to ensure that all healthcare provided via telehealth maintains an equal quality level to that of in-person care. PMID- 19548828 TI - Implementation of telepathology in the republic of georgia. AB - Telepathology in the Republic of Georgia is still evolving. Although much progress has been made around the world, especially in the field of digital imaging and virtual slides, telepathology in Georgia still revolves around static telepathology. The results of the NATO Networking Infrastructure Grant "ePathology-Virtual Pathology Center in Georgia as the continuation of Virtual Health Care Center" are presented. It is a practical implementation of telepathology in Georgia as a best practice example. Using basic methodology: idea-analysis-conception-implementation-test/deployment, the ePathology server was created for the establishment of telepathology in Georgia. Two main services were made available on the server: Simple Machines Forum (eConsultation) and Moodle (eLearning) under the premise "keep it small, safe, and simple." The ePathology server works well. By its application, introduction of the Pap-smear technique and 2001 Bethesda System for reporting cervicovaginal cytologic diagnosis has been done. The application of easily available and adaptable technology, together with the improvement of the infrastructure conditions, is the essential basis for telemedical applications. Telepathology is a very useful and applicable tool for consulting on difficult pathology cases. It has significantly increased knowledge exchange and thereby ensured a better medical service. PMID- 19548829 TI - Can "patient keeper" help in-patients? AB - The aim of this paper is to present our "Patient Keeper" application, which is a client-server medical application. "Patient Keeper" is designed to run on a mobile phone for the client application and on a PC for the server application using J2ME and JAVA2, respectively. This application can help doctors during visits to their patients in hospitals. The client application allows doctors to store on their mobile phones the results of their diagnoses and findings such as temperature, blood pressure, medications, analysis, etc., and send this information to the server via short message service (SMS) for storage in a database. The server can also respond to any request from the client and send the result via Bluetooth, infrared, or over the air. Experimental results showed a significant improvement of the healthcare delivery and reduction for in-patient stay. PMID- 19548831 TI - A dual-fluorescence high-throughput cell line system for probing multidrug resistance. AB - The efflux pump P-glycoprotein (ATP-binding cassette B1, multidrug resistance [MDR] 1, P-gp) has long been known to contribute to MDR against cancer chemotherapeutics. We describe the development of a dual-fluorescent cell line system to allow multiplexing of drug-sensitive and P-gp-mediated MDR cell lines. The parental OVCAR-8 human ovarian carcinoma cell line and the isogenic MDR NCI/ADR-RES subline, which stably expresses high levels of endogenous P-gp, were transfected to express the fluorescent proteins Discosoma sp. red fluorescent protein DsRed2 and enhanced green fluorescent protein, respectively. Co-culture conditions were defined, and fluorescent barcoding of each cell line allowed for the direct, simultaneous comparison of resistance to cytotoxic compounds in sensitive and MDR cell lines. We show that this assay system retains the phenotypes of the original lines and is suitable for multiplexing using confocal microscopy, flow cytometry, or laser scanning microplate cytometry in 1,536-well plates, enabling the high-throughput screening of large chemical libraries. PMID- 19548832 TI - No man is an island: multiple pathologies in patients with malaria. PMID- 19548833 TI - HIV infection, malnutrition, and invasive bacterial infection among children with severe malaria. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection, malnutrition, and invasive bacterial infection (IBI) are reported among children with severe malaria. However, it is unclear whether their cooccurrence with falciparum parasitization and severe disease happens by chance or by association among children in areas where malaria is endemic. METHODS: We examined 3068 consecutive children admitted to a Kenyan district hospital with clinical features of severe malaria and 592 control subjects from the community. We performed multivariable regression analysis, with each case weighted for its probability of being due to falciparum malaria, using estimates of the fraction of severe disease attributable to malaria at different parasite densities derived from cross sectional parasitological surveys of healthy children from the same community. RESULTS: HIV infection was present in 133 (12%) of 1071 consecutive parasitemic admitted children (95% confidence interval [CI], 11%-15%). Parasite densities were higher in HIV-infected children. The odds ratio for admission associated with HIV infection for admission with true severe falciparum malaria was 9.6 (95% CI, 4.9-19); however, this effect was restricted to children aged 1 year. Malnutrition was present in 507 (25%) of 2048 consecutive parasitemic admitted children (95% CI, 23%-27%). The odd ratio associated with malnutrition for admission with true severe falciparum malaria was 4.0 (95% CI, 2.9-5.5). IBI was detected in 127 (6%) of 2048 consecutive parasitemic admitted children (95% CI, 5.2%-7.3%). All 3 comorbidities were associated with increased case fatality. CONCLUSIONS: HIV, malnutrition and IBI are biologically associated with severe disease due to falciparum malaria rather than being simply alternative diagnoses in co-incidentally parasitized children in an endemic area. PMID- 19548835 TI - A retrospective evaluation of completion rates, total cost, and adverse effects for treatment of latent tuberculosis infection in a public health clinic in central massachusetts. AB - Completion rates, total cost, and adverse effects were compared for patients in central Massachusetts treated for latent tuberculosis infection with 9 months of isoniazid or 4 months of rifampin. Although the adverse effects were similar between the 2 groups, 4 months of rifampin was associated with significantly better completion rates and less hepatotoxicity yet higher total cost. PMID- 19548834 TI - Characteristics of O157 versus non-O157 Shiga toxin-producing Escherichia coli infections in Minnesota, 2000-2006. AB - BACKGROUND: Escherichia coli O157:H7 (O157) is the Shiga toxin-producing E. coli (STEC) serotype most frequently isolated and most often associated with hemolytic uremic syndrome (HUS) in the United States. Non-O157 STEC serotypes can also cause serious illness, but their impact as pathogens remains undefined. We compared characteristics of non-O157 and O157 STEC infections identified through sentinel surveillance. METHODS: Sentinel sites included a metropolitan health maintenance organization laboratory and a hospital laboratory serving a small city and rural area. We received sorbitol-MacConkey agar plates from every stool culture performed at both sites during 2000-2006. Colony sweeps were screened for stx1 and stx2 by polymerase chain reaction. E. coli identity, serotype, and presence of stx1 and/or stx2 were confirmed on individual isolates. RESULTS: Two hundred six STEC isolates were identified: 108 (52%) were non-O157 serotypes, and 98 (48%) were O157. Of non-O157 cases, 54% involved bloody diarrhea, and 8% involved hospitalization. Non-O157 isolates with at least stx2 were not more likely to cause severe illness (bloody diarrhea, hospitalization, or HUS) than were non-O157 isolates with only stx1. O157 cases were more likely than non-O157 cases to involve bloody diarrhea (78% vs 54%; P < .001), hospitalization (34% vs 8%; P < .001 and HUS (7% vs 0%; P = .005). When including only isolates with at least stx2, O157 cases were still more likely to involve bloody diarrhea (78% vs 56%; P = .02) and hospitalization (33% vs 12%; P = .01) than non-O157 cases. CONCLUSIONS: Differences in severity among STEC infections could not be explained by stx2, suggesting that additional factors are important in STEC virulence. PMID- 19548836 TI - Long-term acute care hospitals. AB - Long-term acute care hospitals (LTACHs) are health care facilities that admit complex patients with acute care needs (eg, mechanical ventilator weaning, administration of intravenous antibiotics, and complex wound care) for a mean duration of stay of 25 days. LTACHs are different than nursing homes and were initially created in the 1990s in an effort to decrease Medicare costs by facilitating prompt discharge from intensive care units of patients with difficulty weaning mechanical ventilation; however, current admission diagnoses are quite broad. Patients admitted to these facilities have multiple comorbidities and are at risk for colonization with multidrug-resistant organisms. LTACH patients have been shown to have high rates of hospital-acquired infections, including central vascular catheter-associated bloodstream infection and ventilator-associated pneumonia. In addition, LTACHs have been implicated in various regional outbreaks of multidrug-resistant organisms. This review summarizes the limited amount of scientific literature on LTACHs while highlighting their infection control problems, as well as the role LTACHs play on regional outbreaks. PMID- 19548837 TI - A new phylogenetic diversity measure generalizing the shannon index and its application to phyllostomid bats. AB - Protecting biodiversity involves preserving the maximum number and abundance of species while giving special attention to species with unique genetic or morphological characteristics. In balancing different priorities, conservation policymakers may consider quantitative measures that compare diversity across ecological communities. To serve this purpose, a measure should increase or decrease with changes in community composition in a way that reflects what is valued, including species richness, evenness, and distinctness. However, counterintuitively, studies have shown that established indices, including those that emphasize average interspecies phylogenetic distance, may increase with the elimination of species. We introduce a new diversity index, the phylogenetic entropy, which generalizes in a natural way the Shannon index to incorporate species relatedness. Phylogenetic entropy favors communities in which highly distinct species are more abundant, but it does not advocate decreasing any species proportion below a community structure-dependent threshold. We contrast the behavior of multiple indices on a community of phyllostomid bats in the Selva Lacandona. The optimal genus distribution for phylogenetic entropy populates all genera in a linear relationship to their total phylogenetic distance to other genera. Two other indices favor eliminating 12 out of the 23 genera. PMID- 19548838 TI - Mutation accumulation in real branches: fitness assays for genomic deleterious mutation rate and effect in large-statured plants. AB - The genomic deleterious mutation rate and mean effect are central to the biology and evolution of all species. Large-statured plants, such as trees, are predicted to have high mutation rates due to mitotic mutation and the absence of a sheltered germ line, but their size and generation time has hindered genetic study. We develop and test approaches for estimating deleterious mutation rates and effects from viability comparisons within the canopy of large-statured plants. Our methods, inspired by E. J. Klekowski, are a modification of the classic Bateman-Mukai mutation-accumulation experiment. Within a canopy, cell lineages accumulate mitotic mutations independently. Gametes or zygotes produced at more distal points by these cell lineages contain more mitotic mutations than those at basal locations, and within-flower selfs contain more homozygous mutations than between-flower selfs. The resulting viability differences allow demonstration of lethal mutation with experiments similar in size to assays of genetic load and allow estimates of the rate and effect of new mutations with moderate precision and bias similar to that of classic mutation-accumulation experiments in small-statured organisms. These methods open up new possibilities with the potential to provide valuable new insights into the evolutionary genetics of plants. PMID- 19548840 TI - Genetic variation promotes long-term coexistence of Brassica nigra and its competitors. AB - How multiple species coexist in the face of limiting resources remains one of the central questions in ecology. Recent theoretical and empirical studies have documented the importance of evolutionary forces in species coexistence. However, there remains a disconnect between these two approaches, as empirical studies are generally too short to explore long-term coexistence and theoretical studies are rarely specific enough to allow for meaningful comparisons with natural systems. Here I combine field data with simulation modeling to test how a genetic trade off between intra- and interspecific competitive ability alters the long-term coexistence of plant species. In two of the three species combinations tested, coexistence was possible only in models that included evolutionary processes. Additionally, genetic variation and the resultant evolutionary change allowed for coexistence under a much wider range of ecological conditions by both increasing equalizing (neutral) effects and providing a novel evolutionary stabilizing (niche) effect. Biodiversity is declining at both the species and the genetic levels. These results suggest that conserving species diversity may depend critically on our ability to conserve the genetic diversity within species. PMID- 19548839 TI - Bacteriolytic activity in the ejaculate of an insect. AB - The rapid evolution of ejaculate components is considered to be largely driven by sexual selection. Less attention has been paid to the fact that sperm and microorganisms frequently meet; we consequently predict selection for substances that protect a male's ejaculate. We report, for the first time, bacteriolytic activity (lysozyme-like immune activity [LLA]) in the ejaculate of an animal, the common bedbug Cimex lectularius. We also show that in almost half the males LLA in the seminal fluid exceeded LLA in the hemolymph. We detected no antimicrobial peptide activity in seminal fluid. Because lysozymes degrade only bacteria, our results suggest that sperm-microbe interactions are probably important in the evolution of ejaculate components and thereby provide a route for natural selection to account for some of the diversity of seminal components. PMID- 19548841 TI - Inhibition of acidic mammalian chitinase by RNA interference suppresses ovalbumin sensitized allergic asthma. AB - Asthma, a chronic helper T cell type 2-mediated inflammatory disease, is characterized by airway hyperresponsiveness and inflammation. Growing evidence suggests that increased expression of acidic mammalian chitinase (AMCase) may play a role in the pathogenesis of asthma. In the present study, we sought to develop an RNA interference approach to suppress allergic asthma in mice through silencing of AMCase expression. Mice sensitized with ovalbumin (OVA) were intratracheally administered a recombinant adeno-associated virus expressing short hairpin RNA (rAAV-shRNA) against AMCase. In OVA-sensitized mice, the development of allergic symptoms was significantly associated with elevated AMCase expression. After administration of rAAV-shRNA, there was a significant reduction of AMCase expression in the lung and in bronchoalveolar lavage fluid (BALF) cells of sensitized mice. Sensitized mice receiving rAAV-shRNA showed a significant improvement in allergic symptoms, including airway hyperresponsiveness (AHR), eosinophil infiltration, eotaxin, interleukin-13 secretion in BALF, and serum OVA-specific IgE level. Our data suggest the hyperexpression of AMCase in asthma can be suppressed by rAAV-mediated shRNA. Silencing AMCase expression by shRNA may be a promising therapeutic strategy in asthma. PMID- 19548842 TI - Combined strategies for liposome characterization during in vitro digestion. AB - Three types of pyranine (HPTS)-containing liposomes were prepared by high pressure homogenization under optimized conditions. At 37 degrees C, they were 1) fluid-state vesicles made from soybean phosphatidylcholine (SPC), 2) gel-state liposomes made from hydrogenated SPC (HSPC), and 3) solid-disordered membranes obtained from HSPC and cholesterol (HSPC-Chol). These liposome formulations were characterized before, during, and after in vitro digestion, which involved the presence of pH gradients, enzymes, and bile salts. Mean sizes and size distributions of the vesicles were determined by DLS; (31)P-NMR (nuclear magnetic resonance) was used to quantify lyso-PC forms; internal pH was monitored throughout digestion with two different fluorescent pH probes; and changes in bilayer permeability and HPTS encapsulation were determined by size-exclusion chromatography and fluorimetry. Differential scanning calorimetry analysis was also performed in order to study the effect of digestion on HSPC vesicles. SPC liposomes were physically stable during digestion; they presented 8% lyso-forms and an HPTS encapsulation around 85% after in vitro digestion. However, they were extremely permeable to ions, so that the internal pH immediately equilibrated with the bulk pH. HSPC liposomes were the most affected by the digestive process. Even though they were chemically stable, as inferred from the low lyso-PC content, very important changes in their size distribution were observed. A final 50% HPTS leakage was quantified after in vitro digestion. Nevertheless, they were the least permeable to protons under pH gradients. HSPC-Chol vesicles presented intermediate permeability to protons, having their internal pH decreased from approximately 6.8 to 4.6 after 1 hour of incubation at pH 2. This was the most chemically stable formulation and showed the highest encapsulation, even after in vitro digestion. Therefore, HSPC-Chol liposomes would be the most adequate choice for the design of lipid products for oral administration. PMID- 19548843 TI - Evaluation of the Innovance D-DIMER analytical performance. AB - BACKGROUND: Widespread use of D-dimer in recent years has led to the development of a number of new fully automated quantitative D-dimer assays. METHODS: We evaluated the analytical performance of the particle-enhanced immunoturbidimetric assay Innovance D-DIMER (Siemens Medical Solutions) on the Behring Coagulation System (BCS) analyzer. RESULTS: Within-run coefficients of variation (CVs) for samples with low, borderline, slightly, and extremely increased D-dimer concentrations were 2.1%-5.5%, whereas between-run CVs for control samples with low and extremely increased D-dimer were 5.5%-8.4%. The assay exhibited good linearity in the working range between 0.17 mg/L and 5.45 mg/L fibrinogen equivalent units (FEU), with the lower limit of detection of 0.099 mg/L FEU. The upper reference value determined in 40 plasma samples from healthy volunteers was 0.495 mg/L FEU. The results obtained in 457 fresh plasma samples were compared with results obtained with VIDAS D-Dimer Exclusion. Passing and Bablok regression analysis demonstrated highly significant correlation (y=1.370x-0.108, r=0.952, p<0.001). Bland and Altman difference plots demonstrated slightly higher results obtained with Innovance D-DIMER that was more pronounced with increasing values. Very good agreement between both assays was observed (kappa=0.860; 95% confidence interval (CI), 0.811-0.908). CONCLUSIONS: This study demonstrates that Innovance D-DIMER fulfills all analytical requirements for daily routine use. PMID- 19548844 TI - Chromosome 9p21 polymorphism is associated with myocardial infarction but not with clinical outcome in Han Chinese. AB - BACKGROUND: rs1333049 polymorphism on chromosome 9p21 has been shown to affect susceptibility to coronary artery disease (CAD) in Caucasians. This study examined the association of rs1333049 with myocardial infarction (MI), angiographic severity of CAD and clinical outcome after a first acute MI in Han Chinese. METHODS: rs1333049 polymorphism was genotyped in 520 patients with a first acute MI and in 560 controls. The number of angiographically documented diseased coronary arteries (luminal diameter stenosis > or = 50%), echocardiographic left ventricular ejection fraction (LVEF), and major adverse cardiac events (MACE) during follow-up (mean, 29+/-15 months) were recorded. RESULTS: Patients with MI had higher frequencies of the CC genotype (30.0% vs. 20.7%) or C allele (55.5% vs. 46.2%) compared with controls (all p<0.01). rs1333049 polymorphism was strongly associated with MI [odds ratio (OR) 1.48, 95% confidence interval (CI) 1.22-1.79] after adjusting for traditional risk factors. Although longer hospitalization stay was observed in patients with the rs1333049 C allele, this polymorphism was not related to angiographic severity of CAD, LVEF, and occurrence of MACE after MI. CONCLUSIONS: This study demonstrates an association of rs1333049 polymorphism locus on chromosome 9p21 with risk for MI, but not with post-MI prognosis in Han Chinese. PMID- 19548845 TI - Multicenter evaluation of the hemolysis index in automated clinical chemistry systems. AB - BACKGROUND: In vitro hemolysis, the prevailing cause of preanalytical error in routine laboratory diagnostics, might influence the reliability of several tests, affect the quality of the total testing process and jeopardize patient safety. Although laboratory instrumentation is now routinely equipped with systems capable of automatically testing and eventually correcting for hemolysis interference, to our knowledge there are no reports that have compared the efficiency of different analytical platforms for identifying and classifying specimens with hemolysis. METHODS: Serum from a healthy volunteer was spiked with varying amounts of hemolyzed blood from the same volunteer, providing a serum free hemoglobin concentration ranging from 0.0 g/L to 2.0 g/L as measured by the reference cyanmethemoglobin assay. The spiked serum samples were shipped to seven separate laboratories and the hemolysis index (HI) was tested in triplicate on the following analytical platforms: Roche Modular System P (n=4) and Integra 400 Plus (n=1), Siemens Dimension RxL (n=3), ADVIA 2400 (n=1) and ADVIA 1800 (n=1), Olympus AU 680 (n=1) and Coulter DXC 800 (n=1). RESULTS: Satisfactory agreement of HI results was observed among the various analytical platforms, despite a trend toward overestimation by the ADVIA 2400 and 1800. After normalizing results according to the instrument-specific alert value, discrepancies were considerably reduced. All instruments except for the Dimension RxL gave values normalized to the instrument-specific alert value, <1.0 for the sample with 0.048 g/L free hemoglobin, and >1.0 for the sample with 0.075 g/L free hemoglobin. The results of the four Modular System P tests were also highly reproducible among the different facilities. When evaluating instruments that provided quantitative HI results, the mean intra-assay coefficient of variation (CV) calculated for the triplicate determinations was always between 0.1% and 2.7%. CONCLUSIONS: The results of this multicenter evaluation confirm that efficiency of different analytical platforms to correctly identify and classify unsuitable samples is satisfactory. However, more effort should be placed on the standardization of reporting HI. All the instruments that we tested provide either quantitative or qualitative results that are essentially comparable, but which should always be compared with the instrument-specific alert values to harmonize their efficiency. PMID- 19548847 TI - Comparison of fifteen immunoassays for the measurement of serum MUC-1/CA 15-3 in breast cancer patients. AB - BACKGROUND: Quality control results for serum MUC-1/CA 15-3 assays have always shown large discrepancies. METHODS: This multicentre study of 15 methods (labelled M1-M15) measured coded sera from 35 patients with breast cancer without recurrence (group 1), 46 patients at 1st metastasis (group 2), and 39 patients with advanced metastases (group 3). Results were compared using parametric statistics, ANOVA, principal component analysis, and receiver operating characteristic (ROC) curves. RESULTS: Mean MUC-1/CA 15-3 concentrations varied widely (75.1-303.0 U/mL, 24.8%) among methods. The false positive (FP) rate for group 1 was 8/521 (1.5%); for group 2 and group 3 false negative (FN) results were 21/680 (3.1%) and 11/583 (1.9%), respectively. Using the ROC cut-offs, we found no FPs for group 1 and no FNs for group 3. However, group 2 showed 16 FNs. All p-values for Pearson's correlation were <0.0001 between methods, except for M11. When comparing methods using different antibodies, discordance rates reached a maximum of 15.2%. Principal component analysis revealed a grouping of methods using: CanAg monoclonal antibodies (mAbs) (M2, M7 and M12); Centocor/Fujirebio mAbs (M3-M6, M8-M10, M14-M15) and Biomira mAbs (M1 and M13); and Centocor/Fujirebio mAbs (M11). CONCLUSIONS: Results were more consistent among methods using the same antibody type. Principal component analysis showed that antibody type was the strongest determinant of immunoassay results. PMID- 19548846 TI - Variation of barrier permeability for albumin and immunoglobulin G influx into cerebrospinal fluid. AB - BACKGROUND: The aim of the present study was to analyze variations in permeability of albumin and immunoglobulin G (IgG) influx into cerebrospinal fluid (CSF) in a clinical setting. METHODS: In a retrospective intra-individual comparison of CSF samples, we used the IgGIndex and its constituents to indicate alterations in IgG/albumin permeability. RESULTS: We found altered IgGIndices in 25/64 patients (range -25% to +44%), with differently altered QAlb and QIgG values (-69% to +549%), unaltered IgG-Indices in a further 25/64 patients with equally altered QAlb and QIgG values (-46% to +107%), and no parameter alteration in 14/64 patients. Parameter alterations in 25/64 patients indicated that permeability of albumin was changed to different extents than for IgG. It changed in the same direction in 20/25 patients, and the opposite in five patients. In further 25/64 patients, equal QAlb and QIgG alterations indicated equally altered permeabilities and/or altered efflux of the proteins. In 14/64 patients, no alteration in permeability or efflux was seen. CONCLUSIONS: Results revealed surprisingly variable intra-individual changes in permeabilities for albumin and IgG in pathologic as well as normal CSF. Differing changes in permeability indicate that the diffusion paths of the two proteins may react to disturbances independently of each other. The details of the influx permeability for albumin and IgG into CSF illustrate the prospect of a more comprehensive insight into the protein exchange between blood and CSF. PMID- 19548848 TI - HER1-4 protein concentrations in normal breast tissue from breast cancer patients are expressed by the same profile as in the malignant tissue. AB - BACKGROUND: The epidermal growth factor receptor HER2 is overexpressed or amplified in 25%-30% of patients with breast cancer. The mechanism behind HER2 amplification is unknown, but may be a patho-physiological phenomenon caused by continuous stimulation and activation of the HER1-4 system. We have mapped the protein concentrations of HER1-4 in breast cancer tissue, autologous reference tissue, normal breast tissue and serum samples, to see whether non-cancer cells from these patients express a protein profile indicating general activation. METHODS: Tissue samples from malignant and adjacent normal breast tissue (autologous reference tissue) were collected from 118 women consecutively admitted for surgical treatment of breast cancer. In addition, 26 samples of normal breast tissue were collected from healthy women having breast reduction surgery. The tissue samples were homogenized and the proteins extracted. The tissue and serum concentrations of HER1-4 were determined quantitatively using a commercially available enzyme linked immunosorbent assay (ELISA) method. RESULTS: HER1 was down regulated in cancer tissue when compared to autologous reference tissue (p=8 x 10(-6)), while HER2 (p<10(-7)) and HER3 (p=3 x 10(-5)) were up regulated. Comparing autologous reference tissue with normal tissue showed down regulation of HER1 (p=0.122) and up regulation of HER2 (p=10(-6)), HER3 (p<10( 7)) and HER4 (p<10(-7)). Furthermore, we observed that correlations between the receptor combinations HER1-2, HER1-3 and HER1-4 were maintained from normal breast tissue to autologous reference breast tissue, but were lost in cancer tissue. CONCLUSIONS: We suggest that these findings indicate that breast cancer is a systemic disease where the HER1-4 system in autologous reference tissue is continuously activated, thus favoring the subsequent development of cancer. PMID- 19548849 TI - Discontinued drugs in 2008: cardiovascular drugs. AB - This perspective is part of an annual series of papers discussing drugs dropped from clinical development in the previous year. Specifically, this paper focuses on the 16 cardiovascular drugs discontinued in 2008. Information for this perspective was derived from a search of the Pharmaprojects database for drugs discontinued after reaching Phase I-III clinical trials. PMID- 19548850 TI - Peritoneal carcinomatosis of gastrointestinal origin: natural history and treatment options. AB - BACKGROUND: Peritoneal carcinomatosis (PC) is a challenging consequence of certain malignancies, associated with a poor prognosis and limited treatment options. Generally, PC has been treated similarly to metastatic cancers of the primary tumor, but associated with worse outcomes when compared to other sites of metastatic disease from the same primary tumor site. With supportive care alone, the median survival with PC is 3-6 months. More recently, a limited number of centers have reported success with cytoreductive surgery (CS) and hyperthermic intraperitoneal chemotherapy (HIPEC) in a subset of patients with PC, resulting in improved survival compared with historical controls. OBJECTIVES: This paper outlines the natural history of PC, and surgical, chemotherapeutic, and combined modality treatment options, with a focus on PC of colorectal (CRC) and appendiceal origin. RESULTS: At this time, the 'standard' treatment for PC remains incompletely defined. As such, the optimal management strategies for both 'localized' and unresectable disease is unclear. CS + HIPEC is a promising treatment with a significant survival benefit of 10 months over systemic therapy alone demonstrated in a clinical trial of patients with CRC. CONCLUSION: Well designed clinical trials need to continue to be offered to improve care and determine the optimal treatment strategies for PC. PMID- 19548851 TI - Fosfomycin for the treatment of infections caused by Gram-positive cocci with advanced antimicrobial drug resistance: a review of microbiological, animal and clinical studies. AB - BACKGROUND: The advancing antimicrobial drug resistance in Gram-positive cocci complicates the selection of appropriate therapy. The re-evaluation of older antibiotics may prove useful in expanding relevant therapeutic options. OBJECTIVE: We sought to evaluate fosfomycin for the treatment of infections caused by methicillin-resistant staphylococci, vancomycin-resistant enterococci, and penicillin-non-susceptible pneumococci. METHODS: We searched in PubMed, Scopus, and the Cochrane Library for studies evaluating the antimicrobial activity of fosfomycin against the above-mentioned pathogens, or the in vivo or clinical effectiveness of fosfomycin for the treatment of infections caused by these pathogens. RESULTS/CONCLUSIONS: As reported in the identified studies, the susceptibility rate of methicillin-resistant Staphylococcus aureus to fosfomycin was > or = 90% in 12/22, and 50-90% in 7/22 studies; the cumulative susceptibility rate was 87.9% (4240/4892 isolates). The cumulative susceptibility rate of vancomycin-resistant enterococci to fosfomycin was 30.3% (183/604 isolates), and that of penicillin-non-susceptible pneumococci was 87.2% (191/219 isolates). Clinical data show that fosfomycin, primarily in combination regimens, has been associated with clinical success in 28/29 (96.6%) cases of infection (mainly pneumonia, bacteremia, and meningitis) by fosfomycin-susceptible isolates of methicillin-resistant S. aureus. The above data support further research on the role of fosfomycin against infections caused by Gram-positive cocci with advanced antimicrobial drug resistance. PMID- 19548852 TI - Antiangiogenic (metronomic) chemotherapy for brain tumors: current and future perspectives. AB - Significant advances in the diagnosis and treatment of brain tumors have been made through better imaging, surgical techniques and advances in radiation therapy. However, the cure rate for most adult and pediatric brain tumor patients has not mirrored this success. Angiogenesis, the development of neovascularization, provides the required nutrients and oxygen to an expanding tumor and is controlled by a complex balance of proangiogenic cytokines and antiangiogenic factors. A series of new inhibitors of angiogenesis are now in clinical trials. Most of these rely on inhibiting tumor cell-mediated cytokines or blocking the activation of their cognate receptors. Cytotoxic chemotherapy, by contrast, targets dividing cells but can be modulated to attack dividing endothelial cells. This review will focus on the use of low-dose antiangiogenic (also called metronomic) chemotherapy to inhibit endothelial cell function and resultant neovascularization in the treatment of adult and pediatric brain tumors. By examining the biology and preclinical findings that led to the development of antiangiogenic/metronomic chemotherapy, clinical studies have been undertaken that support the role of this approach in the clinic, and have led to the introduction of a number of markers being used to better predict active combinations and appropriate patient populations. PMID- 19548855 TI - Sustained-release fampridine for multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic inflammatory and neurodegenerative disease of the central nervous system of unknown cause in which neurological impairment and disability result from demyelination and axonal loss. Physiologically, myelin loss leads to changes in axonal ion channels that cause conduction failure. Axonal loss leads to a reduction in signal strength in neuronal pathways. Fampridine (4-aminopyridine) is a potassium channel blocker that can increase action potential duration and amplitude, leading to improved conduction in demyelinated nerve fibers and to increased neurotransmitter release at synaptic endings. Fampridine treatment can improve ambulation in some MS patients, but can also cause seizures and other side effects. Pharmacokinetic studies show that improvement in neurological deficits is primarily related to the total fampridine dose, while seizure induction is related to peak serum levels. To reduce side effects, a slow-release (SR) formulation of fampridine was developed. Two Phase III studies of fampridine SR have now shown that treatment can improve leg strength and walking speed in patients with MS; a new drug application has been filed with the US FDA. PMID- 19548854 TI - Prostvac-VF: a vector-based vaccine targeting PSA in prostate cancer. AB - Prostvac is a prostate cancer vaccine regimen consisting of a recombinant vaccinia vector as a primary vaccination, followed by multiple booster vaccinations employing a recombinant fowlpox vector. Both vectors contain the transgenes for prostate-specific antigen (PSA) and multiple T-cell co-stimulatory molecules (TRICOM). The PSA-TRICOM vaccines infect antigen-presenting cells (APCs) and generate proteins that are expressed on the surface of the APCs in an immune context. The interaction of these APCs with T cells initiates a targeted immune response and T cell-mediated tumor cell destruction. Preliminary clinical trials have indicated negligible toxicity, and Phase II trials have suggested a survival benefit after treatment with Prostvac, especially in patients with indolent disease characteristics. Preclinical and clinical data indicate that radiation, hormonal therapy, and chemotherapy may be combined with Prostvac to enhance the vaccine's efficacy. Additional strategies are in development to further enhance the clinical benefits of Prostvac, and a Phase III trial is being planned in metastatic castration-resistant prostate cancer. PMID- 19548853 TI - The development of antibody-IL-2 based immunotherapy with hu14.18-IL2 (EMD 273063) in melanoma and neuroblastoma. AB - Patients with high risk melanoma and neuroblastoma frequently experience recurrence despite surgical resection and appropriate adjuvant therapies. Immunotherapy with the immunocytokine hu14.18-IL2 (EMD-273063) was developed by means of fusion of two molecules of IL-2 to the monoclonal antibody, 14.18, that recognizes GD2, expressed on the earlier mentioned malignancies. This article will discuss the results of preclinical work using EMD-273063 therapy, including data suggesting that intratumoral therapy may have enhanced antitumor benefit compared with intravenous therapy. Initial clinical trials in adult melanoma and pediatric neuroblastoma have demonstrated acceptable toxicity profiles in dosing that induces immune activation. Preclinical and initial clinical data suggest greater efficacy in the setting of minimal residual disease; therefore, future clinical testing is planned to test the benefit of EMD-273063 in this setting. PMID- 19548856 TI - Cixutumumab. AB - The IGF pathway plays a major role in cancer cell proliferation, survival and resistance to antineoplastic therapies in many human malignancies. As such, interference with this pathway is the target of many investigational pharmacologic agents. Cixutumumab, a monoclonal antibody to IGF-1R, utilizes this concept. In this review, we summarize preclinical, pharmacologic and early clinical data regarding this agent and discuss the impact this drug might have on the future treatment of human cancers. PMID- 19548857 TI - Rilpivirine: a novel non-nucleoside reverse transcriptase inhibitor. AB - Combination antiretroviral therapy has transformed the prognosis and life expectancy of HIV-1 infected individuals in resource-rich settings. British guidelines currently recommend the non-nucleoside reverse transcriptase inhibitor (NNRTI) efavirenz as part of first-line treatment in therapy-naive HIV-1 infected individuals. However, efavirenz is limited by its low genetic barrier to the development of resistance, together with its potential for CNS toxicities. To overcome these obstacles, several 'next-generation' NNRTIs are in various stages of clinical development. Here, we review the journey of rilpivirine (also known as TMC278, Tibotec), from the discovery of the chemical compound, through successful Phase I and II development, to its current position of being studied in international Phase III trials for the treatment of therapy-naive HIV-1 infected subjects using a 25 mg daily dose. Pharmacokinetic findings and food and drug interactions are discussed, together with safety profile. Rilpivirine has demonstrated high antiviral activity (including against NNRTI-resistant isolates) in vitro, with similar rates of virological suppression in therapy-naive individuals at 96 weeks when compared to efavirenz. Rilpivirine seems to be well tolerated with less CNS disturbance than efavirenz, and has non-teratogenic potential; however, unfavorable interactions with acid suppressant medications will require heightened vigilance when rilpivirine is used in widespread clinical practice. PMID- 19548858 TI - Tesofensine--a novel potent weight loss medicine. Evaluation of: Astrup A, Breum L, Jensen TJ, Kroustrup JP, Larsen TM. Effect of tesofensine on bodyweight loss, body composition, and quality of life in obese patients: a randomised, double blind, placebo-controlled trial. Lancet 2008;372:1906-13. AB - BACKGROUND: The incidence of obesity is increasing; this is of major concern, as obesity is associated with cardiovascular disease, stroke, type 2 diabetes, respiratory tract disease, and cancer. OBJECTIVES/METHODS: This evaluation is of a Phase II clinical trial with tesofensine in obese subjects. RESULTS: After 26 weeks, tesofensine caused a significant weight loss, and may have a higher maximal ability to reduce weight than the presently available anti-obesity agents. However, tesofensine also increased blood pressure and heart rate, and may increase psychiatric disorders. CONCLUSIONS: It is encouraging that tesofensine 0.5 mg may cause almost double the weight loss observed with sibutramine or rimonabant. As tesofensine and sibutramine have similar pharmacological profiles, it would be of interest to compare the weight loss with tesofensine in a head-to-head clinical trial with sibutramine, to properly assess their comparative potency. Also, as teso fensine 0.5 mg increases heart rate, as well as increasing the incidence of adverse effects such as nausea, drug mouth, flatulence, insomnia, and depressed mode, its tolerability needs to be further evaluated in large Phase III clinical trials. PMID- 19548859 TI - Cyclosporine A for neuroprotection: establishing dosing guidelines for safe and effective use. AB - Numerous neuroprotective compounds have been investigated to ameliorate secondary changes and the progression of injury after the primary insult in traumatic brain injury (TBI). This cascade of events is complex and difficult to abate once initiated following the primary injury. The clinical consequences of this secondary injury are unpredictable and often permanently incapacitating. Cyclosporine A (CsA) interrupts the endogenous mediators of secondary insult through inhibition of the mitochondrial permeability transition pore and prevention of subsequent mitochondrial dysfunction. This drug may have a role in neuroprotection but has several pharmacologic effects that must be considered when using it in the TBI population. This review discusses the physiologic responses following TBI that may affect CsA efficacy and safety when used for neuroprotective indications. So far, CsA seems to be safe in the TBI population. The role of CsA after acute TBI will be better defined after the completion of upcoming planned clinical trials. PMID- 19548860 TI - Type I IFNs and their role in the development of autoimmune diseases. AB - BACKGROUND: Since their initial use in the 1980s, IFNs have become an essential component of the therapy for many diseases such as hepatitis and multiple sclerosis. Although they have been extremely useful in conditions that pose therapeutic challenges, complications associated with their use have been widely reported including emerging reports of several autoimmune diseases. Many of these reports have shed light on the pathogenesis of autoimmune disorders and helped to highlight not only the critical role of type I IFNs in defense against viral infections but also the pivotal role they occupy in the interface between innate and adaptive immunity. Many patients with autoimmune disease have increased responsiveness to type I IFNs (alpha/beta), and therapy with these cytokines has induced or unmasked autoimmune disease in many additional patients. OBJECTIVE: The objective of this paper is to discuss the role of type I IFNs in autoimmunity. METHODS: The literature regarding type I IFNs and autoimmunity was reviewed using the Medline database from 1950 to 2009. Search terms included 'interferon alpha' and 'autoimmune disease' and 'interferon beta' and 'autoimmune disease'. Case reports, case series, reviews and prospective studies were included in the analysis. RESULTS/CONCLUSIONS: In the literature a variety of autoimmune disorders have reportedly been induced by the use of type I IFNs, being used, although these are primarily in the form of case reports and case series. Nevertheless, there is a growing body of molecular evidence to support the clinical association. The role of IFNs in the induction of autoimmunity is complex with interplay of many genetic and environmental factors that influence the balance between normal and aberrant immune responsiveness, ultimately leading to the observed clinical manifestations. PMID- 19548861 TI - Recent developments in the use of oral anticoagulants. AB - For many years, vitamin K antagonists, unfractionated heparins, low-molecular weight heparins and a pentasaccharide were the only anticoagulant drugs available for the prevention of venous thromboembolism after surgery. However, their benefits were associated with disadvantages, such as their subcutaneous route of administration or the need for coagulation monitoring. Research was challenged to develop new drugs that would simplify thromboprophylaxis while showing equivalent or better efficacy. Rivaroxaban and dabigatran are now available in some countries for the prevention of venous thromboembolism after total hip or knee replacement in adults. Apixaban is also undergoing trials for this indication. Additionally, these drugs show potential for stroke prevention in atrial fibrillation, treatment of venous thromboembolism and prevention of secondary events in acute coronary syndrome. PMID- 19548862 TI - Proteasome inhibitors in cancer therapy. AB - The ubiquitin-proteasome pathway is involved in intracellular protein turnover and plays an essential function in many cellular processes, thus contributing to cellular homeostasis. The observation that proteasome inhibitors are able to induce apoptosis preferentially in tumor cells opened the way to their use as potential drugs. One of these drugs, bortezomib, was introduced in cancer therapy and its use was approved for the treatment of multiple myeloma and mantle cell lymphoma. The investigation of the mechanisms through which bortezomib causes cell death of cancer cells showed that this drug affects many cellular pathways and many of its effects cannot be related to proteasome inhibition. These observations, as well as the occurrence of bortezomib resistance observed in some myeloma patients treated with this drug, prompted the study of new proteasome inhibitors. These studies have lead to a new generation of proteasome inhibitors, some of them exhibiting the property of selective proteasome inhibition and efficacy in cancer therapy without inducing the side effects elicited by bortezomib. The ensemble of these drugs offers the scenario of the possible development of a family of proteasome inhibitors, contributing a drug arsenal for the therapy of cancer and other diseases. PMID- 19548863 TI - Biomarkers of aging with prognostic and predictive value in non-oncological diseases. AB - In this review we discuss the current knowledge on Biomarkers of Aging (BoAs) in the context of human diseases and their value as predictive or prognostic markers. The vast majority of studies using BoAs in a clinical context have been undertaken by determining telomere length in peripheral blood mononuclear cells (PBMCs), whereas the expression of cell cycle inhibitors and an increase in advanced glycation end products (AGEs) have rarely been used. Here we summarize the impact of BoAs on non-oncological, hematological, cardiovascular, metabolic, renal and neurological diseases, as well as on overall survival. The specific methodologies utilized are described and evaluated for their high-throughput potential. PMID- 19548864 TI - Potential therapeutic effects of Na+/Ca2+ exchanger inhibition in cardiac diseases. AB - The Na+/Ca2+ exchanger (NCX) has a pivotal role in cardiac Na+ and Ca2+ homeostasis and is an essential pathway for Ca2+ extrusion from cardiomyocytes. Altered NCX function may result in abnormal Ca2+ release from the sarcoplasmic reticulum (SR) and impaired cardiac electrical activity and contractility in several diseases, like arrhythmias, ischemia/reperfusion injury, hypertrophy and heart failure. This review focuses on the role of the exchanger and the major effects of partial NCX blockade in normally functioning and diseased hearts. In healthy cardiac cells consequences of a partial NCX blockade were found to be moderate and species dependent. In rabbit and dog practically no change in the magnitude of the calcium transients and mechanical activity was observed, while elevation of systolic calcium levels and a concomitant positive inotropic action were demonstrated in rat and murine hearts. On the other hand, NCX inhibition represents a novel potential therapeutic strategy in case of a variety of cardiac dysfunctions. Partial NCX blockade was shown to have beneficial effects in animal models of ischemia/reperfusion injury and also antiarrhythmic and positive inotropic actions in the failing heart. Further progress in this field is seriously hampered by the absence of really selective NCX inhibitors. KB-R7943 and SEA0400 are widely used NCX blockers, both drugs, however, have limited selectivity and efficacy, properties required to initiate relevant clinical trials. In summary, there is an increasing demand by both researchers and clinicians for new NCX inhibitors with significantly enhanced selectivity and functionality. PMID- 19548865 TI - Use and safety of calcium channel blockers in obstetrics. AB - Calcium-Channel Blockers (CCBs), or calcium antagonists, are a heterogeneous group of drugs that produce cardiovascular effects by preventing the influx of calcium ions through L-type voltage-dependent calcium channels in specialized electrical system and conduction tissue cells, like myocardial and vascular smooth muscle cells. In recent years, CCBs have found their way in obstetrics and gynecology, especially in the management of hypertensive disorders of pregnancy and preterm labor. The lack of adequate data had created uncertainty about the safety of CCBs in pregnancy. Teratogenicity with these agents has been demonstrated in animals, but no cases of possible human malformation or deformity have been reported. Data from human studies suggest that CCBs may cause a clinically insignificant fall in maternal mean arterial pressure, but have little to no effect on uterine perfusion. In many countries, CCBs remain unlicensed for use in pregnancy and it is unlikely the manufacturers will ever apply for this status to change. We do believe that this is the situation of CCBs for those critical second- and third-trimester conditions such as Gestational hypertension, Preeclampsia, HELLP syndrome and preterm labor. PMID- 19548866 TI - Doxorubicin: the good, the bad and the ugly effect. AB - The anthracycline doxorubicin (DOX) is widely used in chemotherapy due to its efficacy in fighting a wide range of cancers such as carcinomas, sarcomas and hematological cancers. Despite extensive clinical utilization, the mechanisms of action of DOX remain under intense debate. A growing body of evidence supports the view that this drug can be a double-edge sword. Indeed, injury to nontargeted tissues often complicates cancer treatment by limiting therapeutic dosages of DOX and diminishing the quality of patients' life during and after DOX treatment. The literature shows that the heart is a preferential target of DOX toxicity. However, this anticancer drug also affects other organs like the brain, kidney and liver. This review is mainly devoted to discuss the mechanisms underlying not only DOX beneficial effects but also its toxic outcomes. Additionally, clinical studies focusing the therapeutic efficacy and side effects of DOX treatment will be discussed. Finally, some potential strategies to attenuate DOX-induced toxicity will be debated. PMID- 19548867 TI - Translocator protein ligands as promising therapeutic tools for anxiety disorders. AB - The Translocator protein (TSPO), formerly known as the peripheral-type benzodiazepine receptor, is an 18 kDa mitochondrial protein primarily involved in steroid biosynthesis in both peripheral and glial cells. It has been extensively reported that TSPO regulates the rate-limiting translocation of cholesterol from the outer to the inner mitochondrial membrane before its transformation by cytochrome P450(scc) into pregnenolone, which is further converted into an array of different steroids. In the brain, neurosteroids such as allopregnanolone and pregnenolone, acting as positive modulators of gamma-aminobutyric type A (GABA(A)) receptors, exert anxiolytic activity. Specific ligands targeting TSPO increase neurosteroid production and for this reason they have been suggested to play an important role in anxiety modulation. Unlike benzodiazepines (Bzs), which represent the most common anti-anxiety drugs administered around the world, selective TSPO ligands have shown anxiolytic effects in animal models without any of the side effects associated with Bzs. Therefore, specific TSPO ligands that are able to promote neurosteroidogenesis may represent the future of therapeutic treatment of anxiety disorders. Furthermore, TSPO expression levels are altered in several different psychiatric disorders in which anxiety is the main symptom. This article reviews the primary and patent literature over the last decade concerning the development of novel TSPO ligands that have resulted effective in various models of anxiety, taking into special consideration their structure activity relationships. PMID- 19548868 TI - Thyroid hormone antagonists: potential medical applications and structure activity relationships. AB - Thyroid hormone receptors (TRs) exert profound effects on development, metabolism, and multiple specific organ functions. Principally by regulating crucial genes in a variety of tissues, the thyroid hormones, 3,5,3'-triiodo-L thyronine (L-T(3), 1) and 3,5,3',5'-tetraiodo-L-thyronine (L-T(4), 2), influence basal calorigenesis and oxygen consumption, cardiac rate and contractility, lipid metabolism, bone structure and strength, and central nervous system functions critical for normal mentation and mood. Elevated levels of circulating and tissue 1 and/or 2 result in the thyrotoxic clinical state, manifested by weight loss despite increased caloric intake; heat intolerance due to increased calorigenesis; cardiac tachyarrhythmias, systolic hypertension, and heart failure; skeletal muscle weakness; and a spectrum of neuropsychiatric symptoms ranging from anxiety to delirium and psychosis. The current standard treatments of endogenous hyperthyroidism causing thyrotoxicosis reduce the overproduction of thyroid hormones by pharmacologically inhibiting their synthesis or release (e.g., with thionamides or lithium, respectively), or by ablating thyroid tissue surgically or with radioiodine. TR-antagonists could hypothetically have significant clinical use in treating thyrotoxic states if they were capable of promptly and completely restoring euthyroid levels of thyroid-specific gene activity. No TRalpha-selective ligands have been prepared up to this date, ligands that potentially would further ameliorate the problem with cardiac disease connected with hyperthyroidism and maybe cardiac arrhythmia. Despite its significant potential use, no TR-antagonist has reached clinical application. Design of TR-antagonists ligands has been based on the attachment of a large extension group at the 5-prime position of 1 or other structurally related analogues. This extension is believed to distort folding of the C-terminal helix (helix 12) to the body of the ligand binding domain (LBD), which normally forms a coactivator site. Examples of synthetic TR antagonists based on this extension strategy are reviewed, as well as other strategies to achieve functional TR antagonism. PMID- 19548869 TI - Blocking receptor protein tyrosine phosphatase beta/zeta: a potential therapeutic strategy for Parkinson's disease. AB - striatum of rodents in experimental models of Parkinson's disease. Interestingly, immunohistochemical studies have shown increased levels of PTN expression in the substantia nigra of patients with Parkinson's disease. Since, in other contexts, PTN has been shown to be critical in repair processes in the injured nervous system, the antecedents suggest that PTN could exhibit protective effects in Parkinson's disease. This hypothesis was confirmed when PTN was shown to support survival of dopaminergic neurons and to promote the differentiation of neural stem cells to dopaminergic neurons. These findings suggest a new therapeutic approach in the treatment of Parkinson's disease based on the molecular mechanism of action of PTN. Pleiotrophin receptor, receptor protein tyrosine phosphatase (RPTP) beta/zeta, is found active in monomeric form in neurons and glia within the central nervous system. Pleiotrophin induces dimerization of RPTPbeta/zeta inactivating its phosphatase activity, thus increasing the phosphorylation levels of its substrates such as beta-catenin, Fyn and beta-adducin. These substrates have been shown to be critical for the proliferation of dopaminergic progenitors and the survival and differentiation of dopaminergic neurons. This review summarizes the strong scientific basis to consider blocking RPTPbeta/zeta as a potentially novel therapeutic strategy in the treatment of Parkinson's disease and discusses various starting points to design antagonists of this receptor. PMID- 19548871 TI - Prenylated isoflavonoids: Botanical distribution, structures, biological activities and biotechnological studies. An update (1995-2006). AB - In contrast with the parent class of flavonoids, the distribution of the isoflavonoid class in the plant kingdom is relatively limited, probably owing to the sporadic occurrence of isoflavone synthase. Isoflavonoids have been mostly found in the subfamily Fabaceae/Papilionoideae of the Leguminosae family. Isoprenoid-substituted (also called complex) isoflavonoids are expressed from a smaller number of plants, as a result of the similarly restricted distribution of prenyltransferases (PT-ase). After the reviews of Tanara & Ibrahim (1995), Boland & Donnelly (1997), the Handbook of Flavonoids by Harborne & C ( Handbook of Flavonoids, 1999), and the paper by Harborne and Williams (2000) few other reports concern the distribution and the biological activity of complex isoflavonoids, except a list of isoflavonoids produced from non leguminous plants. This review deals with an update of the literature on isoprenylated isoflavonoids in the years 1995-2006 and is focused on the following highlights. 1. Natural sources of complex isoflavonoids (2000-2006); 2. Chemical structure variety: new entries (2000-2006) 3. Biological activities and a possible structure-activity relationship (1995-2006) 4. In vitro production and microbial metabolism (1995-2006). PMID- 19548870 TI - New therapeutic targets for drug design against Trypanosoma cruzi, advances and perspectives. AB - Chagas disease is one of the most important parasitic diseases in Latin America, affecting 16 to 18 million people. Nifurtimox and Benznidazol are drugs that are commonly used in its treatment; however, these drugs produce several adverse reactions and are not effective in the chronic phase of the disease. Therefore, the design, synthesis, and biological evaluation of new compounds with potential activity against Trypanozoma cruzi are of great importance. We review six proteins involved in the biochemical metabolism of Trypanosoma cruzi that have recently been studied as potential targets for designing new drugs for Chagas disease. These are farnesyl pyrophosphate synthase, trans-sialidase, cruzain cystein protease, trypanothione reductase, glucose 6-phosphate-dehydrogenase, glyceraldehyde 3-phosphate-dehydrogenase, and alpha-hydroxy acid dehydrogenase. We also review the advances of compounds recently designed based on structure activity, and the perspectives of new compounds that inhibit these therapeutic targets. PMID- 19548872 TI - Recent developments in the chemistry of thebaine and its transformation products as pharmacological targets. AB - The most practical synthetic routes to the preparation of as important pharmaceuticals as oxycodone, naloxone, naltrexone, nalbuphine and buprenorphine have utilized the alkaloid, thebaine, as a starting material. This review intends to focus on chemical transformations of morphinans which resulted in morphinandiene derivatives with well-established and novel pharmacological potencies. These chemical transformations were mainly associated with the formation and substitution of the unique diene structure of the ring C of the morphinan backbone. PMID- 19548873 TI - A new drug candidate (GEMSP) for multiple sclerosis. AB - GEMSP is a mixture of functional polypeptides: fatty acids linked to poly-L Lysine (PL), antioxidants linked to PL, free radical scavengers linked to PL, and amino acids linked to PL (patent numbers 6114388 (USA) and 792167 (EU)). In this review, we update the data on this new drug reported in the literature. There is evidence suggesting that GEMSP is a good candidate for the treatment of multiple sclerosis (MS), an inflammatory and neurodegenerative disease of the central nervous system characterized by focal leukocyte inflammation, demyelization and axonal degeneration, resulting in nerve cell dysfunction. Experimental autoimmune encephalomyelitis (EAE) is the main animal model used in the study of MS, a T cell-mediated autoimmune disease of the central nervous system. EAE has many clinical and histopathological similarities to MS. In this model, preclinical studies on GEMSP have demonstrated that the drug strongly inhibits brain leukocyte infiltration and completely abolishes EAE episodes and clinical scores, and it also appears that GEMSP preserves myelin integrity. In general, treatment with the free constituents of GEMSP (not linked to the inert carrier protein) is poorly active against brain leukocyte infiltration in EAE-immunized animals. This means that free molecules (not linked to PL) exert a very poor action on such infiltration and that these molecules are either rapidly incorporated into the metabolism or are degraded. Moreover, with immunocytochemical techniques, it has been demonstrated that one component of GEMSP, the methionine compound, is stored inside the motoneurons of the ventral horn of the spinal cord. However, this component of GEMSP has not been found in the brain. The new candidate for MS therapy has shown no toxicity either in experimental animals or in humans. An open clinical trial in humans has demonstrated that GEMSP is completely safe. In addition, the approved drugs for the treatment of MS exert marked side effects, but no side effects have been reported following the administration of GEMSP. The results obtained at six months of treatment with low doses of GEMSP (0.75 mg/day) in that open clinical trial in humans were as follows: 55% of the patients maintained a stable expanded disability status scale (EDSS) value and 18% of the patients had a decreased EDSS value instead of a normal progression of 0.25 point on the mean EDSS scale. We focus our review on the following topics: 1) EAE models and clinical evaluation; 2) the synthesis of GEMSP; 3) the effects of GEMSP dosage on EAE; 4) the effects of GEMSP on brain leukocyte infiltration; 5) GEMSP inside motoneurons; 6) the role of the components of GEMSP; and 7) GEMSP in MS patients, GEMSP toxicity, and side effects. In conclusion, all the data reported indicate that GEMSP is a new potential drug candidate for the treatment of MS. PMID- 19548875 TI - Receptor dependent multidimensional QSAR for modeling drug--receptor interactions. AB - Quantitative Structure Activity Relationship (QSAR) is an approach of mapping chemical structure to properties. A significant development can be observed in the last two decades in this method which originated from the Hansch analysis based on the logP data and Hammett constant towards a growing importance of the molecular descriptors derived from 3D structure including conformational dynamics and solvation scenarios. However, molecular interactions in biological systems are complex phenomena generating extremely noisy data, if simulated in silico. This decides that activity modeling and predictions are a risky business. Molecular recognition uncertainty in traditional receptor independent (RI) m-QSAR cannot be eliminated but by the inclusion of the receptor data. Modeling ligand receptor interactions is a complex computational problem. This has limited the development of the receptor dependent (RD) m-QSAR. However, a steady increase of computational power has also improved modeling ability in chemoinformatics and novel RD QSAR methods appeared. Following the RI m-QSAR terminology this is usually classified as RD 3/6D-QSAR. However, a clear systematic m-QSAR classification can be proposed, where dimension m refers to, the static ligand representation (3D), multiple ligand representation (4D), ligand-based virtual or pseudo receptor models (5D), multiple solvation scenarios (6D) and real receptor or target-based receptor model data (7D). PMID- 19548874 TI - The L-type Ca(2+) channel as a therapeutic target in heart disease. AB - The L-type Ca(2+) channel plays a critical role in cardiac function as the main route for entry of calcium into cardiac myocytes. It is essential to excitability as it shapes the long plateau phase of the cardiac action potential that is unique to cardiac ventricular myocytes. It is necessary for contraction as it triggers the release of calcium from sarcoplasmic reticulum stores for actin myosin interaction. The L-type Ca(2+) channel also regulates cytoplasmic calcium levels. It is well recognised that an increase in intracellular calcium is involved in the activation of growth-promoting signal pathways. Recently reactive oxygen species have been implicated in the activation of signal pathways and the development of pathological hypertrophy. There is now evidence that implicates activation of the L-type Ca(2+) channel with persistent alterations in calcium homeostasis and cellular reactive oxygen species production as a possible trigger of cardiac hypertrophy. A number of different approaches have been used to modify channel function with the view to preventing ischemia-reperfusion injury, cardiac hypertrophy or cardiac failure providing good evidence that the L-type Ca(2+) channel may be an efficacious target in the prevention of cardiac pathology. PMID- 19548878 TI - Inhibitory effects of kynurenic acid, a tryptophan metabolite, and its derivatives on cytosolic sulfotransferases. AB - KYNA (kynurenic acid) is an endogenous metabolite of tryptophan in the kynurenine pathway and has been characterized as an antagonist of ionotropic glutamate receptors. In addition, we have reported this endogenous compound as a potent inhibitor of SULTs (cytosolic sulfotransferases). In the present study we characterized the inhibitory effects of KYNA on several human (h) and mouse (m) recombinant SULTs. No sulfate metabolite of KYNA was detected with mouse and human SULTs examined under the conditions used, suggesting that it is a bona fide inhibitor of SULTs. Among the mouse enzymes examined, KYNA exhibited selective inhibitory effects on Sult1b1-mediated sulfation of various compounds with IC50 values in the low micromolar range (2.9-4.9 microM). KYNA also exerted an inhibitory activity towards hSULT1A1 and hSULT1B1. The inhibitory potency of KYNA for mSult1b1 was stronger than that of 2,6-dichloro-4-nitrophenol, a known non specific SULT inhibitor, whereas the potencies of these two inhibitors for hSULT1B1 were comparable. The inhibitory characteristics of KYNA were clearly distinct from those of mefenamic acid, a selective inhibitor of SULT1A enzymes. The KYNA derivatives 5,7-dichlorokynurenic acid and L689,560 exhibited preferential inhibitory effects on hSULT1A1 and hSULT1B1 respectively. Interestingly, gavestinel, another KYNA derivative, was found to be an extremely potent inhibitor of hSULT1B1. Finally, we have demonstrated that the mechanism underlying the KYNA inhibition varied depending on the enzyme and substrate involved. Taken together, the present results unveil another distinct aspect of KYNA and its derivatives as an inhibitor of SULTs. PMID- 19548879 TI - Recruitment of NAADP-sensitive acidic Ca2+ stores by glutamate. AB - NAADP (nicotinic acid-adenine dinucleotide phosphate) is an unusual second messenger thought to mobilize acidic Ca(2+) stores, such as lysosomes or lysosome like organelles, that are functionally coupled to the ER (endoplasmic reticulum). Although NAADP-sensitive Ca(2+) stores have been described in neurons, the physiological cues that recruit them are not known. Here we show that in both hippocampal neurons and glia, extracellular application of glutamate, in the absence of external Ca(2+), evoked cytosolic Ca(2+) signals that were inhibited by preventing organelle acidification or following osmotic bursting of lysosomes. The sensitivity of both cell types to glutamate correlated well with lysosomal Ca(2+) content. However, interfering with acidic compartments was largely without effect on the Ca(2+) content of the ER or Ca(2+) signals in response to ATP. Glutamate but not ATP elevated cellular NAADP levels. Our results provide evidence for the agonist-specific recruitment of NAADP-sensitive Ca(2+) stores by glutamate. This links the actions of NAADP to a major neurotransmitter in the brain. PMID- 19548880 TI - Two novel phosphatidylinositol-4-phosphate 5-kinase type Igamma splice variants expressed in human cells display distinctive cellular targeting. AB - The generation of various phosphoinositide messenger molecules at distinct locations within the cell is mediated via the specific targeting of different isoforms and splice variants of phosphoinositide kinases. The lipid messenger PtdIns(4,5)P(2) is generated by several of these enzymes when targeted to distinct cellular compartments. Several splice variants of the type Igamma isoform of PIPK (PtdIns4P 5-kinase), which generate PtdIns(4,5)P(2), have been identified, and each splice variant is thought to serve a unique functional role within cells. Here, we have identified two novel C-terminal splice variants of PIPKIgamma in human cells consisting of 700 and 707 amino acids. These two splice variants are expressed in multiple tissue types and display PIPK activity in vitro. Interestingly, both of these novel splice variants display distinct subcellular targeting. With the addition of these two new splice isoforms, there are minimally five PIPKIgamma splice variants that have been identified in mammals. Therefore, we propose the use of the HUGO (Human Genome Organization) nomenclature in the naming of the splice isoforms. PIPKIgamma_i4 (700 amino acids) is present in the nucleus, a targeting pattern that has not been previously observed in any PIPKIgamma splice variant. PIPKIgamma_i5 (707 amino acids) is targeted to intracellular vesicle-like structures, where it co localizes with markers of several types of endosomal compartments. As occurs with other PIPKIgamma splice variants, the distinctive C-terminal sequences of PIPKIgamma_i4 and PIPKIgamma_i5 may facilitate association with unique protein targeting factors, thereby localizing the kinases to their appropriate cellular subdomains for the site-specific generation of PtdIns(4,5)P(2). PMID- 19548881 TI - Anterior chamber depth and iridocorneal angle in healthy White subjects: effects of age, gender and refraction. AB - PURPOSE: Prior to phakic intraocular lens implantation, it is important to obtain precise knowledge of the anterior chamber depth (ACD). Accurate topographic evaluation of the iridocorneal angle is helpful in estimating risk for angle closure glaucoma. This study investigated the use of the Orbscan II system to measure ACD and the iridocorneal angle in healthy subjects and assessed the influences of age, gender and spherical equivalent on these parameters. METHODS: The Orbscan II system was used to determine the ACD and iridocorneal angle in eight different positions in 390 healthy White subjects with a mean age of 41+/- 16years (range 10-80 years). The sample included 242 male and 148 female subjects. The influences of age, gender and spherical equivalent were assessed using multiple regression analysis. RESULTS: Mean ACD was 2.87 +/- 0.04 mm in male subjects and 2.81+/-0.37mm in female subjects. The explanatory variables relevant to the ACD were age (partial regression coefficient B = -0.0115, p < 0.0001), spherical equivalent (B = - 0.0562, p< 0.0001) and gender (B = 0.0996, p=0.0036). The mean iridocorneal angle (MIA) was 30.7 +/- 2.0 degrees in male and 31.6 +/- 2.1 degrees in female subjects. The variables relevant to the MIA were gender (B =- 0.865, p < 0.0001), age (B =- 0.017, p = 0.0007) and spherical equivalent (B = - 0.121, p = 0.001). The superior iridocorneal angle displayed the strongest negative correlation with age, whereas the temporal angle exhibited the least correlation with age. CONCLUSIONS: The decline in ACD appears to be linear with age, amounting to a mean of 0.58 mm over 50 years. This may become clinically relevant in the use of phakic intraocular lenses. In addition, the angle is more severely constricted in the superior quadrant than in the temporal quadrant. PMID- 19548882 TI - Early changes in clear cornea incision after phacoemulsification: an anterior segment optical coherence tomography study. AB - PURPOSE: We aimed to investigate the ultrastructures of clear corneal incisions 24 hours after phacoemulsification using anterior segment optical coherence tomography (AS-OCT). METHODS: Sixty eyes of 60 patients scheduled for cataract surgery were randomly selected. All eyes underwent the same phacoemulsification procedure carried out by one experienced surgeon. Two-plane temporal clear corneal tunnel incisions were performed. The same types of intraocular lens and implant system were used in all patients. Images of the ultrastructures of the corneal incisions were taken using AS-OCT before and 1 day after surgery. RESULTS: Corneal thickness increased at the incision site after surgery in all eyes (p < 0.001). Epithelial bulla in the incision region was seen in two eyes (3%). Gaping at the internal aspect of the corneal wound was seen in 42 eyes (70%). Eyes with gaping had thicker localized cornea (p = 0.002). Descemet's membrane detachment was seen in 49 eyes (82%); this seemed to be associated with lower preoperative intraocular pressure (p = 0.01). CONCLUSIONS: Anterior segment OCT provides sensitive and detailed measurements of the ultrastructures in clear corneal incision. PMID- 19548883 TI - Variation in dispersal mortality and dispersal propensity among individuals: the effects of age, sex and resource availability. AB - 1. Dispersal of individuals between habitat patches depends on both the propensity to emigrate from a patch and the ability to survive inter-patch movement. Environmental factors and individual characteristics have been shown to influence dispersal rates but separating the effects of emigration and dispersal mortality on dispersal can often be difficult. In this study, we use a soil mite laboratory system to investigate factors affecting emigration and dispersal mortality. 2. We tested the movement of different age groups in two-patch systems with different inter-patch distances. Differences in immigration among age groups were primarily driven by differences in emigration but dispersal mortality was greater for some groups. Immigration declined with increasing inter-patch distance, which was due to increasing dispersal mortality and decreasing emigration. 3. In a second experiment, we compared the dispersal of recently matured males and females and tested the impact of food availability during the developmental period on their dispersal. Dispersal was found to be male biased but there was no significant sex bias in dispersal mortality. There was some evidence that food availability could affect emigration and dispersal mortality. 4. These results demonstrate that both emigration and dispersal mortality can be affected by factors such as individual age and resource availability. Understanding these effects is likely to be important for predicting the fitness costs and population consequences of dispersal. PMID- 19548884 TI - Detection of different quorum-sensing signal molecules in a virulent Edwardsiella tarda strain LTB-4. AB - AIMS: The aim of this study was to elucidate the potential quorum-sensing (QS) signal molecules of an emerging pathogen (Edwardsiella tarda strain LTB-4) of cultured turbot (Scophthalmus maximus). METHODS AND RESULTS: A sensitive and rapid double-layer plate method using biosensor strain Agrobacterium tumefaciens KYC55 was developed to detect the N-acylhomoserine lactone (AHL)-related compounds in bacteria. LTB-4 was found to have two QS systems, one was based on the AHLs and the other was based on the autoinducer-2 (AI-2). The AI-2 activity produced by LTB-4 was growth phase dependent and topped at OD(600) of 1.0. The protocol to detect cholerae autoinducer 1 (CAI-1) activity in bacteria was modified, lowering the background luminescence of biosensor strain Vibrio harveyi JAF375. CAI-1 activity could not be detected in LTB-4. CONCLUSION: Edwardsiella tarda LTB-4 produced at least four kinds of AHLs during its whole growth phase. In comparison with the AHL-inducing QS, AI-2 may be the first predominant signal, functioning at early exponential phase. LTB-4 did not produce any CAI-1 activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Different QS signal molecules of Edw. tarda LTB-4 were clarified by improved bioassays. In contrast to earlier studies detecting two types of AHLs, strain LTB-4 produced at least four kinds of AHLs, which seemed to be C(4)-HSL, C(6)-HSL, 3-oxo-C(6)-HSL and an uncharacterized AHL molecule. PMID- 19548885 TI - Fluoroquinolone resistance in clinical and environmental isolates of Escherichia coli in Mexico City. AB - AIMS: To assess the different phenotypes and mechanisms of fluoroquinolone (FQ) resistance in clinical and environmental isolates of Escherichia coli. METHODS AND RESULTS: We compared FQ-resistant E. coli isolates, measuring minimal inhibitory concentrations (MIC) of ciprofloxacin, along with susceptibility to other antibiotics. We also searched for the presence of efflux pumps, using efflux inhibitors, and for plasmid-borne FQ-resistance by PCR. We found that, aside from the higher FQ-resistance prevalence among clinical strains, environmental ones resist much lower concentrations of ciprofloxacin. Efflux pumps mediate fluoroquinolone resistance as frequently among environmental isolates than in clinical strains. Plasmid-borne qnrA genes were not detected in any resistant strain. CONCLUSIONS: Environmental FQ-resistant strains may have a nonclinical origin and/or a selective pressure different from the clinical use of FQs. SIGNIFICANCE AND IMPACT OF THE STUDY: The identification of the source of low-level FQ-resistant strains (ciprofloxacin MIC c. 8 microg ml(-1)) in the environment could be important to curb the rapid emergence and spread of FQ resistance in clinical settings, as these strains can easily become fully resistant to FQ concentrations achievable in fluids and tissues during therapy. PMID- 19548886 TI - A novel in vitro flat-bed perfusion biofilm model for determining the potential antimicrobial efficacy of topical wound treatments. AB - AIMS: To develop an in vitro flat-bed perfusion biofilm model that could be used to determine the antimicrobial efficacy of topically applied treatments. METHODS AND RESULTS: Pseudomonas aeruginosa and Staphylococcus aureus biofilms were grown within continuously perfused cellulose matrices. Enumeration of the biofilm density and eluate was performed at various sampling times, enabling determination of the biofilm growth rate. Two antimicrobial wound dressings were applied to the surface of mature biofilms and periodically sampled. To enable real-time imaging of biofilm growth and potential antimicrobial kinetics, a bioluminescent Ps. aeruginosa biofilm was monitored using low-light photometry. Target species produced reproducible steady-state biofilms at a density of c. 10(7) per biofilm support matrix, after 24-h perfusion. Test dressings elicited significant antimicrobial effects, producing differing kill kinetic profiles. There was a good correlation between photon and viable count data. CONCLUSIONS: The model enables determination of the antimicrobial profile of topically applied treatments against target species biofilms, accurately differentiating bactericidal from bacteriostatic effects. Moreover, these effects could be monitored in real time using bioluminescence. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first in vitro biofilm model which can assess the antimicrobial potential of topical therapies in a dynamic growth environment. PMID- 19548887 TI - Antimicrobial and antistaphylococcal biofilm activity from the sea urchin Paracentrotus lividus. AB - AIMS: Staphylococcal biofilm-associated infections are resistant to conventional antibiotics. Consequently, new agents are needed to treat them. With this aim, we focused on the effector cells (coelomocytes) of the sea urchin Paracentrotus lividus immune system. METHODS AND RESULTS: We tested the activity of the 5-kDa peptide fraction of the cytosol from coelomocytes (5-CC) against a group of Gram positive, Gram-negative bacteria and fungi. We determined minimal inhibitory concentrations (MICs) ranging from 253.7 to 15.8 mg ml(-1). We observed an inhibitory activity and antibiofilm properties of 5-CC against staphylococcal biofilms of reference strains Staphylococcus epidermidis DSM 3269 and Staphylococcus aureus ATCC 29213. The antimicrobial efficacy of 5-CC against the biofilms of clinical strain Staph. epidermidis 1457 was also tested using live/dead staining in combination with confocal laser scanning microscopy. At a sub-MIC concentration (31 x 7 mg ml(-1)) of 5-CC the formation of young (6-h old) and mature (24-h old) staphylococcal biofilms was inhibited. CONCLUSIONS: The biological activity of 5-CC could be attributed to three peptides belonging to the sequence segment 9-41 of a beta-thymosin of P. lividus. SIGNIFICANCE AND IMPACT OF THE STUDY: The effector cells of P. lividus represent an interesting source of marine invertebrates-derived antimicrobial agents in the development of new strategies to treat staphylococcal biofilms. PMID- 19548888 TI - Bacterial community associated with ensilage process of wilted guinea grass. AB - AIMS: To determine the effects of wilting, storage period and bacterial inoculant on the bacterial community and ensiling fermentation of guinea grass silage. METHODS AND RESULTS: Fermentation products, colony counts and denaturing gradient gel electrophoresis (DGGE) profiles were determined. There was more lactic acid than acetic acid in all silages, but the lactic acid to acetic acid ratio decreased with storage time. This shift from lactic to acetic acid was not prevented even with a combination of wilting and bacterial inoculant. The DGGE analyses suggest that facultatively heterofermentative lactic acid bacteria (Lactobacillus plantarum, Lactobacillus brevis and Lactobacillus pentosus) were involved in the shift to acetic acid fermentation. CONCLUSIONS: Lactic acid can dominate the fermentation in tropical grass silage with sufficient wilting prior to ensiling. Prolonged storage may lead to high levels of acetic acid without distinctive changes in the bacterial community. SIGNIFICANCE AND IMPACT OF THE STUDY: The bacterial community looks stable compared to fermentation products over the course of long storage periods in tropical grass silage. Acetic acid fermentation in tropical grass silage can be a result of the changes in bacterial metabolism rather than community structure. PMID- 19548889 TI - Quantification of Lawsonia intracellularis in porcine faeces by real-time PCR. AB - AIM: To develop and to validate a method for the quantification of Lawsonia intracellularis in porcine faeces by real-time PCR. METHODS AND RESULTS: A real time PCR including a calibrator based on plasmid DNA for quantification by means of DeltaDeltaCt method was evaluated. The parameters specificity, detection limit, quantification limit, linearity, range, repeatability, precision and recovery were validated. The detection limit of the agent was 1 copy per reaction, and quantification was reliable between 10(1) and 10(7) copies per microl reaction volume. The linearity calculated by logistic regression revealed a slope of -3.329 reflecting an efficiency of 99.7% for the assay. Moreover, it was shown that storage of samples and repetition of tests including DNA isolation by same or other investigators did not influence the outcome. CONCLUSION: The quantification method described herein revealed consistent results for the quantitation of L. intracellularis in porcine faeces samples. SIGNIFICANCE AND IMPACT OF THE STUDY: In contrast to common PCR in combination with gel electrophoresis, this validated quantification method based on real-time PCR enhances a reliable quantification and is even more sensitive. PMID- 19548890 TI - Lactobacillus fermentum BCS87 expresses mucus- and mucin-binding proteins on the cell surface. AB - AIMS: To identify and characterize adhesion-associated proteins in the potential probiotic Lactobacillus fermentum BCS87. METHODS AND RESULTS: Protein suspensions obtained from the treatment of Lact. fermentum BCS87 with 1 mol 1(-1) LiCl were analysed by Western blotting using HRP-labelled porcine mucus and mucin. Two adhesion-associated proteins with relative molecular weight of 29 and 32 kDa were identified. The N-terminal and internal peptides of the 32 kDa protein (32-Mmubp) were sequenced, and the corresponding gene (32-mmub) was found by inverse polymerase chain reaction. The complete nucleotide sequence of 32-mmub revealed an open reading frame of 903 bp encoding a primary protein of 300 amino acids and a mature protein of 272 residues. A basic local alignment search showed 47-99% identity to solute-binding components of ATP binding cassette transporter proteins in Lactobacillus, Streptococcus and Clostridium. An OpuAC-conserved domain was identified and phylogenetic relationship analysis confirmed that 32 Mmubp belongs to the OpuAC family. CONCLUSIONS: Adhesion of Lact. fermentum BCS87 appeared to be mediated by two surface-associated proteins. 32-Mmubp is a component of ABC transporter system that also functions as an adhesin. SIGNIFICANCE AND IMPACT OF THE STUDY: Characterization of 32-Mmubp and 32-mmub will contribute to understanding the host-bacteria interactions of Lact. fermentum with the intestinal tract of pigs. PMID- 19548891 TI - Deradelphous cephalothoracopagus in kittens. AB - Cephalothoracopagus is a very rare form of conjoined twins and is characterized by fusion of heads and thoraxes with two separate spines, limbs and pelves. The aim of this study was to describe a case of female cephalothoracopagus kitten puppy. The most important gross findings involved the external body and some of the internal organs. Radiological features revealed main developmental abnormality of the head, spines and thorax. Authors discuss the pathogenic mechanisms of this condition, infrequently reported in veterinary practice, pointing out the importance of embryonic duplications commonly associated with dystocia. PMID- 19548892 TI - Characterization of an organic-solvent-tolerant Brevibacillus agri strain 13 able to stabilize solvent/water emulsion. AB - Brevibacillus agri strain 13 was isolated and characterized as a Gram-positive organic-solvent-tolerant bacterium able to grow at 45 degrees C. It can tolerate high concentrations (5% and 20%, v/v) of various organic solvents with a broad range of log P(ow) when the organic solvent was provided as a nonaqueous layer. Although it can tolerate a number of aromatic solvents, it cannot utilize them as a sole carbon source. The surface characteristics of cells exposed to organic solvent were investigated using the bacterial adhesion to hydrocarbon test, a contact angle measurement, zeta potential determination, and fluorescence microscopy analysis and compared with that of nonexposed cells. The results showed that although it has a hydrophilic cell surface, it has a unique indigenous cell surface characteristic in which the cells can stabilize solvent in-water emulsion by adhering to the solvent-water interface of the solvent droplets. The tolerance and predilection of B. agri strain 13 toward organic solvents may suggest its potential application as a whole-cell biocatalyst for the biotransformation process of water-immiscible substrate(s). PMID- 19548893 TI - Phylogeny and in situ identification of a novel gammaproteobacterium in activated sludge. AB - Failure of a continuously aerated sequencing batch reactor (SBR) pilot plant enhanced biological phosphorus removal (EBPR) process, designed to remove phosphorus from the clarified effluent from a conventional non-EBPR wastewater treatment plant, was associated with the dominance (c. 50% of the biovolume) of gammaproteobacterial coccobacilli. Flow cytometry and subsequent clone library generation from an enriched population of these Gammaproteobacteria showed that their 16S rRNA genes were most similar to partial clone sequences obtained from an actively denitrifying SBR community, and from anaerobic : aerobic EBPR communities. Under the SBR operating conditions used here, these cells stained for poly-beta-hydroxyalkanoates, but never polyphosphate. Applying FISH probes designed against them in combination with microautoradiography showed that they could also assimilate acetate 'aerobically'. FISH analyses of biomass samples from the full-scale treatment plant providing the pilot plant feed showed that they were present there in high numbers. However, they were not detected by FISH in laboratory-scale communities of the same aerated laboratory-scale EBPR process even when EBPR had failed, or from several full-scale EBPR plants or other activated sludge processes. PMID- 19548894 TI - Proteome analysis of outer membrane vesicles from a clinical Acinetobacter baumannii isolate. AB - The secretion of outer membrane vesicles (OMVs) is one of the major mechanisms by which Gram-negative bacteria deliver effector molecules to host cells. Acinetobacter baumannii is an important opportunistic pathogen in hospital acquired infections, but the secretion system for effector molecules to induce host cell damage has not been characterized. In the present study, we investigated the secretion of OMVs from a clinical A. baumannii isolate and analyzed the comprehensive proteome of A. baumannii-derived OMVs. Acinetobacter baumannii secreted OMVs into the extracellular milieu during in vitro growth. Using 1-DE and LC-MS/MS protein identification and assignment analysis, 132 different proteins associated with OMVs were identified. These proteins were derived from outer membranes (n=26), periplasmic space (n=6), inner membranes (n=8), cytoplasm (n=43), and unknown localization or multiple localization sites (n=49) according to the cell location prediction programs. Among the proteins associated with OMVs, a potent cytotoxic molecule, outer membrane protein A, was highly enriched and several putative virulence-associated proteins were also identified. These results suggest that OMVs from A. baumannii are an important vehicle designed to deliver effector molecules to host cells. PMID- 19548895 TI - Comparison of quantitative and molecular genetic variation of native vs. invasive populations of purple loosestrife (Lythrum salicaria L., Lythraceae). AB - Study of adaptive evolutionary changes in populations of invasive species can be advanced through the joint application of quantitative and population genetic methods. Using purple loosestrife as a model system, we investigated the relative roles of natural selection, genetic drift and gene flow in the invasive process by contrasting phenotypical and neutral genetic differentiation among native European and invasive North American populations (Q(ST) - F(ST) analysis). Our results indicate that invasive and native populations harbour comparable levels of amplified fragment length polymorphism variation, a pattern consistent with multiple independent introductions from a diverse European gene pool. However, it was observed that the genetic variation reduced during subsequent invasion, perhaps by founder effects and genetic drift. Comparison of genetically based quantitative trait differentiation (Q(ST)) with its expectation under neutrality (F(ST)) revealed no evidence of disruptive selection (Q(ST) > F(ST)) or stabilizing selection (Q(ST) < F(ST)). One exception was found for only one trait (the number of stems) showing significant sign of stabilizing selection across all populations. This suggests that there are difficulties in distinguishing the effects of nonadaptive population processes and natural selection. Multiple introductions of purple loosestrife may have created a genetic mixture from diverse source populations and increased population genetic diversity, but its link to the adaptive differentiation of invasive North American populations needs further research. PMID- 19548896 TI - Association between human cases and poultry outbreaks of highly pathogenic avian influenza in Vietnam from 2003 to 2007: a nationwide study. AB - This study quantifies the spatio-temporal association between outbreaks of highly pathogenic avian influenza H5N1 in domestic poultry (n = 3050) and human cases (n = 99) in Vietnam during 2003-2007, using rare events logistic regression. After adjusting for the effect of known confounders, the odds of a human case being reported to authorities increased by a factor of 6.17 [95% confidence interval (CI) 3.33-11.38] and 2.48 (95% CI 1.20 - 5.13) if poultry outbreaks were reported in the same district 1 week and 4 weeks later respectively. When jointly considering poultry outbreaks in the same and neighbouring districts, occurrence of poultry outbreaks in the same week, 1-week later, and 4 weeks later increased the odds of a human case by a factor of 2.75 (95% CI 1.43-5.30), 2.56 (95% CI 1.31-5.00) and 2.70 (95% CI 1.56-4.66) respectively. Our study found evidence of different levels of association between human cases and poultry outbreaks in the North and the South of the country. When considering the 9-week interval extending from 4 weeks before to 4 weeks after the week of reporting a human case, in the South poultry outbreaks were recorded in 58% of cases in the same district and 83% of cases in either the same or neighbouring districts, whereas in the North the equivalent results were only 23% and 42%. The strength of the association between human and poultry cases declined over the study period. We conclude that owner reporting of clinical disease in poultry needs to be enhanced by targeted agent-specific surveillance integrated with preventive and other measures, if human exposure is to be minimized. PMID- 19548897 TI - Analysis of antigenic cross-reactivity between subgroup C avian pneumovirus and human metapneumovirus by using recombinant fusion proteins. AB - Avian pneumovirus subgroup C (APV/C) has recently been reported to be more closely related to human metapneumovirus (hMPV) as determined by sequence analysis. To examine the antigenic relationship between APV/C and hMPV, the APV/C fusion (F) gene was cloned and expressed as an uncleaved glycoprotein in a baculovirus system. The reactivity of the APV/C F protein with antibodies against APV subgroups A, B, C, and hMPV was examined by Western blot analysis. The results showed that the expressed APV/C F protein was not only recognized by APV/C-specific antibodies but also by antibodies raised against hMPV. Previously expressed recombinant hMPV F protein also reacted with APV/C-specific antibodies, suggesting that there was significant antigenic cross-reactivity and a potential evolutionary relationship between hMPV and APV/C. Interestingly, the recombinant F proteins from APV/C and hMPV were not recognized by polyclonal antibodies specific to APV subgroups A and B. PMID- 19548898 TI - Endemic transmission of Rift Valley Fever in Senegal. AB - Rift Valley fever (RVF) is an expanding zoonotic disease transmitted from ruminant to ruminant by Culicidae mosquitoes. In 2004, a longitudinal serological survey was performed on small ruminants in the Ferlo are (Senegal) to study RVF transmission and compared the results with those obtained from the same study in 2003. The results confirm that the disease is endemic and that the spatial transmission of RVF is highly heterogeneous. The virus could be maintained during dry season by transovarian transmission in Aedes vexans. Further studies are needed to improve the understanding of the epidemiological cycle of RVF in this region to implement adapted surveillance measures. PMID- 19548899 TI - Immunostimulation to reduce recurrence after surgery for anal condyloma acuminata: a prospective randomized controlled trial. AB - AIM: Human papillomavirus is the most common cause of sexually transmitted disease. It is associated with immunosuppression and shows a marked tendency to recur. We investigated a natural immunostimulant aimed to reduce recurrence. METHOD: A randomized controlled study was carried out including 261 patients allocated to surgical excision alone (control group; n = 122) and surgical excision plus postoperative immunostimulation for 30 days with a natural product (STET; study group; n = 139). Patients with HIV positivity were excluded. All patients gave fully informed consent. RESULTS: The patients were followed for 6 months after surgery. Recurrence occurred in 7.2% (10/139) in the study group and in 27.1% (33/122) in the control group (P < 0.0001). There were no significant differences in the sex, sexual orientation, number of lesions, time to diagnosis and treatment or localization of lesions in the two groups. CONCLUSIONS: Immunostimulation using a natural product significantly reduced the incidence of recurrence of anal condylomata in patients undergoing surgical excision. PMID- 19548900 TI - Evaluation of gadolinium-enhanced T1-weighted magnetic resonance imaging in the preoperative assessment of local staging in rectal cancer. AB - AIM: The aim of this study was to determine whether gadolinium-enhanced T1 weighted magnetic resonance (MR) sequence is beneficial in the preoperative assessment of tumour and nodal staging in patients with primary rectal cancer. METHOD: Eighty-eight patients with primary rectal cancer underwent preoperative MR imaging, followed by surgical resection. Two radiologists independently reviewed (i) T2-weighted MR images (T2WI); (ii) gadolinium-enhanced T1-weighted MR images (T1 + Gd); (iii) MR combined with T2WI and T1 + Gd for the prediction of tumour and nodal stage compared with histopathologic findings as the end point. Differences in the diagnostic performance of T2WI only, T1 + Gd image only and combined T2WI and T1 + Gd MR images were analyzed by comparing areas under receiver operating characteristic curves (Az) for each reader. Interobserver agreement was also calculated. RESULTS: There was no significant difference in the Az values of T2WI only, T1 + Gd image only and combined T2WI and T1 + Gd images for the prediction of tumour staging (Az of T2WI, T1 + Gd and combined MR images for reader 1, 0.80, 0.76 and 0.85; reader 2, 0.83, 0.82 and 0.87) and nodal staging (Az for reader 1, 0.73, 0.73 and 0.81; reader 2, 0.79, 0.80 and 0.83). Interobserver agreement for the prediction of tumour staging was moderate to substantial, while only fair agreement was noted for the prediction of nodal staging. CONCLUSION: Gadolinium-enhanced T1-weighted MRI did not increase the diagnostic yield for tumour and nodal staging, and may be omitted in the MR protocol for preoperative assessment of primary rectal cancer. PMID- 19548901 TI - Stapled haemorrhoidopexy for IV degree haemorrhoids. PMID- 19548902 TI - A possible troponin I leak arising from bowel preparation for elective colonoscopy - are there more implications than observed? PMID- 19548903 TI - Mild and moderate factor VIII deficiency: inhibitor risks. PMID- 19548904 TI - Intensive peri-operative use of factor VIII and the Arg593-->Cys mutation are risk factors for inhibitor development in mild/moderate hemophilia A. AB - BACKGROUND: A severe and challenging complication in the treatment of hemophilia A is the development of inhibiting antibodies (inhibitors) directed towards factor VIII (FVIII). Inhibitors aggravate bleeding complications, disabilities and costs. The etiology of inhibitor development is incompletely understood. OBJECTIVES: In a large cohort study in patients with mild/moderate hemophilia A we evaluated the role of genotype and intensive FVIII exposure in inhibitor development. PATIENTS/METHODS: Longitudinal clinical data from 138 mild/moderate hemophilia A patients were retrospectively collected from 1 January 1980 to 1 January 2008 and analyzed by multivariate analysis using Poisson regression. RESULTS: Genotyping demonstrated the Arg593Cys missense mutation in 52 (38%) patients; the remaining 86 patients had 26 other missense mutations. Sixty-three (46%) patients received intensive FVIII concentrate administration, 41 of them for surgery. Ten patients (7%) developed inhibitors, eight of them carrying the Arg593Cys mutation. Compared with the other patients, those with the Arg593Cys mutation had a 10-fold increased risk of developing inhibitors (RR 10; 95% CI, 0.9-119).The other two inhibitor patients had the newly detected mutations Pro1761Gln and Glu2228Asp. In both these patients and in five patients with genotype Arg593Cys, inhibitors developed after intensive peri-operative use of FVIII concentrate (RR 186; 95% CI, 25-1403). In five of the 10 inhibitor patients FVIII was administered by continuous infusion during surgery (RR 13; 95% CI, 1.9 86). CONCLUSION: The Arg593Cys genotype and intensive peri-operative use of FVIII, especially when administered by continuous infusion, are associated with an increased risk for inhibitor development in mild/moderate hemophilia A. PMID- 19548905 TI - The active conformation of von Willebrand factor in patients with thrombotic thrombocytopenic purpura in remission. AB - BACKGROUND: Functional deficiency of ADAMTS13 in thrombotic thrombocytopenic purpura (TTP) patients is associated with circulating ultralarge von Willebrand factor (VWF) molecules that display spontaneous platelet-binding capacities. Upon remission, however, ADAMTS13 activity does not always return to baseline. OBJECTIVE: To study ADAMTS13 and VWF-related features in TTP patients in remission. METHODS: ADAMTS13 activity, anti-ADAMTS13 antibodies, VWF antigen, ultralarge VWF and levels of VWF that circulate in a glycoprotein Ibalpha-binding conformation were determined in plasma samples of 22 acquired TTP patients in remission between 1 month and 6 years after achieving remission. The composition of active multimers was investigated with a novel immunoprecipitation assay based on monoclonal antibody AU/VWF-a12, which specifically recognizes the active conformation of VWF. RESULTS: ADAMTS13 activity was undetectable in 23% of the patients, even years after they had achieved remission, and lack of ADAMTS13 activity was associated with increased active VWF levels and the presence of ultralarge VWF multimers. Active VWF levels and ultralarge VWF were also associated with blood groups. Results from immunoprecipitation experiments revealed the full range of multimers to be present. CONCLUSION: ADAMTS13 deficiency and the concurrent presence of ultralarge VWF and increased active VWF levels can be detected in TTP patients for years after they have achieved remission. Immunoprecipitation results suggest that the active conformation of VWF may be present in the lower molecular weight multimers, but future studies are necessary to confirm our findings. PMID- 19548906 TI - Three residues at the interface of factor XI (FXI) monomers augment covalent dimerization of FXI. AB - BACKGROUND: Human plasma factor XI is a homodimer, with each monomer comprising a catalytic domain and four homologous 'apple' domains. The monomers bind to each other through non-covalent bonds and through a disulfide bond between Cys321 residues in apple 4 domains. OBJECTIVE: To identify residues essential for dimerization in the FXI monomer interface. METHODS: Specificity-determining residues in apple 4 domains were sought by sequence alignment of FXI and prekallikrein apple domains in different species. Specific residues identified in apple 4 domains were mutagenized and expressed in baby hamster kidney (BHK) cells for evaluation of their effect on FXI dimerization, analyzed by non-reduced sodium dodecylsulfate polyacrylamide gel electrophoresis and size-exclusion chromatography. RESULTS: Among the 19 residues of the FXI monomer interface, Leu284, Ile290 and Tyr329 were defined as specificity-determining residues. Substitutions of these residues or pairs of residues did not affect FXI synthesis and secretion from transfected BHK cells, but did impair dimerization, despite the presence of cysteine at position 321. The double mutant 284A/290A yielded predominantly a monomer, whereas all other single or double mutants yielded monomers as well as disulfide-bonded dimers. CONCLUSIONS: The data suggest that Leu284, Ile290 and Tyr329 in the interface of FXI monomers are essential for forming non-covalently bonded dimers that facilitate formation of a disulfide bonded stable FXI dimer. PMID- 19548907 TI - Variability of platelet aggregate dispersal with glycoprotein IIb-IIIa antagonists eptifibatide and abciximab. AB - BACKGROUND: Utilization of glycoprotein IIb-IIIa (GPIIb-IIIa) inhibitors improves outcomes of patients with acute coronary syndromes (ACS), including those undergoing percutaneous coronary intervention (PCI). These results may be related to the ability of the inhibitors to destabilize coronary thrombi, reduce microembolization, and restore vessel patency. OBJECTIVE: To evaluate in vitro the ability of GPIIb-IIIa antagonists, abciximab and eptifibatide, to promote the disaggregation of platelet-rich thrombus. METHODS: Antagonist-induced disaggregation was assayed in plasma by aggregometry, as well as in whole blood by point of care and capillary perfusion systems. Fibrinogen dissociation from the platelet surface was quantified by flow cytometry. RESULTS: Significant disaggregation of 5 microm ADP-induced aggregates was observed after addition of either agent. The maximum extent and rate of disaggregation were significantly higher with eptifibatide than with abciximab. Both antagonists also dispersed 2 microg mL(-1) collagen-induced aggregates, again with eptifibatide having a greater effect. Eptifibatide, but not abciximab (up to 10 microg mL(-1)), was efficient at dissociating aggregates to single platelets in whole blood and dispersing aggregates that had been aged for 30 min before treatment. Eptifibatide also reduced existing thrombus burden in the perfusion model under arterial flow conditions. A key mechanism of aggregate dispersal was antagonist induced displacement of platelet-bound fibrinogen, which was greater with eptifibatide, a competitive inhibitor of fibrinogen binding, than with the noncompetitive inhibitor, abciximab. CONCLUSIONS: These results suggest that drug concentration and residence time, along with thrombus extent and age, may be critical determinants in promoting timely recanalization. PMID- 19548908 TI - N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine induces apoptosis through the activation of caspases-3 and -8 in human platelets. A role for endoplasmic reticulum stress. AB - BACKGROUND: Apoptosis or programmed cell death involves a number of biochemical events, including the activation of caspases, which lead to specific cell morphology changes and ultimately cell death. Traditionally, two apoptotic pathways have been described: the cell-surface death receptor-dependent extrinsic pathway and the mitochondria-dependent intrinsic pathway. Alternatively, apoptosis has been reported to be induced by endoplasmic reticulum (ER) stress, which is mainly induced by a reduction in intraluminal free Ca(2+) concentration ([Ca(2+)](ER)). OBJECTIVES: The present study aimed to investigate the development of apoptotic events after ER stress induced by N,N,N',N'-tetrakis(2 pyridylmethyl)ethylenediamine (TPEN), an ER Ca(2+) chelator, in human platelets. METHODS: Changes in cytosolic free Ca(2+) concentration, caspase activity and phosphatidylserine externalization were determined by fluorimetric techniques. RESULTS: Our results indicate that TPEN reduces the amount of free Ca(2+) releasable by the Ca(2+)-mobilizing agonist thrombin. TPEN induced activation of caspase-3, -8 and -9 and subsequent phosphatidylserine externalization. The ability of TPEN to induce phosphatidylserine externalization was smaller than that of thrombin. In addition, TPEN was able to induce phosphorylation of the eukaryotic initiation factor 2 alpha (eIF2 alpha). TPEN-mediated caspase-3 activation requires functional caspase-8, but is independent of H(2)O(2) generation. Activation of caspase-3 and -8 by TPEN was prevented by salubrinal, an agent that prevents ER stress-induced apoptosis. CONCLUSION: These findings provide experimental evidence for the existence of ER stress-mediated apoptosis in human platelets, a process that might limit platelet life span upon prolonged stimulation with agonists. PMID- 19548910 TI - Beneficial effect of clopidogrel in a mouse model of polymicrobial sepsis. PMID- 19548911 TI - Faster platelet recovery by high-dose dexamethasone compared with standard-dose prednisolone in adult immune thrombocytopenia: a prospective randomized trial. PMID- 19548909 TI - Differential stimulation of monocytic cells results in distinct populations of microparticles. AB - BACKGROUND: Microparticles (MPs), small vesicles shed from stimulated cells, permit cross-talk between cells within a particular environment. Their composition is thought to reflect their cell of origin, and differs according to whether they are produced by stimulation or by apoptosis. Whether MP properties vary according to stimulus is not yet known. METHODS: We studied the characteristics of MPs produced from monocytic THP-1 cells upon stimulation with lipopolysaccharide or a soluble P-selectin chimera, using proteomics, flow cytometry, western blotting, and electron microscopy. RESULTS: Utilizing a novel criterion of calcein-AM staining to define MPs, we found that MP populations were similar with respect to size, presence and organization of cytoskeleton, and expression of certain antigens. The MPs shared the same level of procoagulant activity. We found that MPs also have distinct characteristics, depending on stimuli. These include differences in phosphatidylserine expression and expression of proteins from specific subcellular locations such as the mitochondria, and of unique antigens such as leukocyte-associated immunoglobin like-receptor (LAIR)-1, which was found only upon stimulation with the soluble P selectin chimera. CONCLUSION: We found that the properties of MPs depend on the stimulus that produced them. This supports the concept that monocytic MPs differentially modulate thrombosis, inflammation and immune regulation according to stimulus. PMID- 19548912 TI - Upper vs. lower extremity deep vein thrombosis: outcome definitions of venous thromboembolism for clinical predictor rules or risk factor analyses in hospitalized patients. PMID- 19548915 TI - Characteristics, head shape measurements and developmental delay in 287 consecutive infants attending a plagiocephaly clinic. AB - AIM: To describe the characteristics, developmental status and severity of head shape deformation in infants presenting to a plagiocephaly clinic. METHODS: Head shape was measured and neck function assessed in 287 consecutive infants presenting to a plagiocephaly outpatient clinic. Information was obtained on demographic and obstetric factors, plagiocephaly history and current positioning strategies. Development was assessed by the Ages and Stages Questionnaire (ASQ). RESULTS: After clinical examination, craniosynostosis was suspected in seven infants (2%) and a skull computed tomography (CT) scan was performed; five showed suture abnormalities. Fifty-eight percent of cases had a history of limitation of neck function; this was more likely in plagiocephalic infants than brachycephalic infants. Males, firstborn infants, instrument-delivered infants, supine sleep position and right-sided flattening were predominant. One or more delays on the ASQ were seen in 36% of infants. CONCLUSION: Physical examination of infants with head shape deformities is essential in order to rule out craniosynostosis. Infants with deformational plagiocephaly frequently have neck muscle dysfunction. We postulate that the higher than expected number of developmental delays may be related to the effects of supine sleep position, low or variable tone, lower activity levels, male gender and neck muscle dysfunction. PMID- 19548917 TI - Nodal marginal zone lymphoma: current knowledge and future directions of an heterogeneous disease. AB - Nodal marginal zone lymphoma (NMZL) is a defined, separate clinicopathological entity. NMZL is a B-cell neoplasm originated in the lymph node, whose histology resembles the nodal infiltration by mucosa-associated lymphoid tissue- or splenic type marginal zone lymphoma, in the absence of clinical evidence of extranodal or spleen disease. The lack of characteristic phenotypic or molecular diagnostic findings is still hampering the reproducibility of this diagnosis. Herein, we review the main morphological and immunophenotypical markers, discussing the differential with other overlapping entities. We also summarize the clinical features and outcome of this rare lymphoma and we discuss the possible association with hepatitis C virus infection. PMID- 19548916 TI - Endometrial population of oestrogen receptors alpha and beta and progesterone receptors A and B during the different phases of the follicular wave of llamas (Lama glama). AB - The aim of this study was to characterize the distribution of oestrogen receptor (ER)alpha and ERbeta as well as both progesterone receptors isoforms progesterone receptor (PR) A and PRB in the luminal and glandular epithelia and stroma of the endometrium during the different phases of the follicular wave in llamas. Six llamas were examined by transrectal ultrasonography, and a transcervical biopsy was obtained when a follicle at the growing, plateau and regressing phase was recorded. Blood samples were collected at the time of biopsy for hormone determinations. An immunohistochemical technique was used to study receptor populations. Total positive area was evaluated in the different cell types by Image Analysis. Mean diameter measurements of the largest follicle were 6.9, 8.5 and 5.1 mm (p < 0.001) and mean plasma oestradiol-17beta concentrations were 27.9 +/- 3.26; 30.0 +/- 2.79 and 24.0 +/- 1.78 pmol/l (p = 0.32) during the growing, plateau and regressing phases, respectively. Immunostaining of ERalpha was higher in the luminal epithelium during the plateau and regressing phases (p < 0.05) than during the growing phase. More positive cells to ERbeta were observed in the glandular epithelium of the growing and plateau phases (p < 0.05) than during the regressing phase. A higher percentage of cells positive to PRB was recorded in the luminal and glandular epithelia during the plateau phase (p < 0.05), while the PRA immunostaining was similar among phases. In brief, this study showed an increased population of ERalpha and PRB in the luminal epithelium, and only of PRB in the glandular epithelium at the time when an ovulatory follicle is present. The physiological importance of these changes in llamas remains to be elucidated. PMID- 19548918 TI - Diffuse large B-cell lymphoma presenting with osteolytic lesions in the bilateral femur. PMID- 19548919 TI - Platelet counts and haemorrhagic diathesis in patients with myelodysplastic syndromes. AB - OBJECTIVES: Most patients with myelodysplastic syndromes (MDS) present with single or multiple lineage cytopenias in peripheral blood despite a hypercellular bone marrow. Thrombocytopenia, attributable to ineffective platelet production by dysfunctional megakaryocytes, has been estimated to occur in 40-65% of patients. However, there are hardly any studies on the clinical relevance of low platelet counts in MDS. METHODS: We retrospectively analysed data from 2900 patients in the Duesseldorf MDS Registry who were diagnosed at our laboratory between 1982 and 2007. RESULTS: At the time of diagnosis, 43% of the patients had a platelet count lower than 100 000/microL. Platelets were lower than 20 000/microL in 7% of the patients, especially in those with advanced stages of MDS, who showed a higher frequency of thrombocytopenia and platelet transfusion dependency. On multivariate analysis, platelet anisometry, hypocellularity of megakaryopoiesis, maturational defects of megakaryocytes and platelets <20 000/microL were independent variables showing a statistically significant correlation (P < 0.05) with clinical signs of bleeding. Platelets lower than 100 000/microL were associated with significantly shortened survival (P < 0.00005), because of an increased risk of progression to acute myeloid leukaemia (AML) (30% vs. 21%) (P < 0.02) and bleeding (16% vs. 8%) (P = 0.0005). CONCLUSIONS: Thrombocytopenia is a strong predictor of short survival, with or without haemorrhagic complications. PMID- 19548920 TI - Bloodstream infections in a medical-surgical intensive care unit: incidence, aetiology, antimicrobial resistance patterns of Gram-positive and Gram-negative bacteria. AB - In the present study, the incidence and antimicrobial resistance patterns of the microorganisms that caused bloodstream infections (BSIs) in a medical-surgical intensive care unit during the years 2005-2007 were determined. The mean BSI incidence density was 6.56 per 1000 patient-days. The incidence density increased linearly during the study period (from 3.57 to 9.60 per 1000 patient-days). Staphylococcus aureus was most frequently isolated (47.3%), followed by Enterococcus spp. (10.8%) and Candida spp. (10.1%). There was a high rate of resistance to several of the prescribed antimicrobials among the bacteria isolated from patients with BSIs. PMID- 19548921 TI - Detection of, and frequent co-infection with, human bocavirus in faecal specimens from children in Wuhan, China. AB - A novel parvovirus, human bocavirus (HBoV), was first discovered in children with respiratory tract infections in Sweden. A retrospective study of HBoV in faecal samples from children suffering from diarrhea, covering a 3-year period (November 2000 to October 2003) in Wuhan, China, was undertaken. PCR assays were used to evaluate 214 faecal samples and to determine the role of HBoV in diarrhoea. Among 196 virus-infected children with diarrhoea, 2.55% were HBoV-positive; however, all HBoV-positive patients were co-infected with common enteric viruses. This result does not support the notion that HBoV is a viral agent causing acute diarrhoea. PMID- 19548922 TI - Sporadic occurrence of CMY-2-producing multidrug-resistant Escherichia coli of ST complexes 38 and 448, and ST131 in Norway. AB - Clinical isolates of Escherichia coli with reduced susceptibility to oxyimino cephalosporins and not susceptible to clavulanic acid synergy (n = 402), collected from Norwegian diagnostic laboratories in 2003-2007, were examined for the presence of plasmid-mediated AmpC beta-lactamases (PABLs). Antimicrobial susceptibility testing was performed for beta-lactam and non-beta-lactam antibiotics using Etest and Vitek2, respectively. The AmpC phenotype was confirmed using the boronic acid test. PABL-producing isolates were detected using ampC multiplex-PCR and examined by bla(AmpC) sequencing, characterization of the bla(AmpC) genetic environment, phylogenetic grouping, XbaI- pulsed-field gel electrophoresis (PFGE), multi-locus sequence typed (MLST), plasmid profiling and PCR-based replicon typing. For the PABL-positive isolates (n = 38), carrying bla(CMY-2) (n = 35), bla(CMY-7) (n = 1) and bla(DHA-1) (n = 2), from out- (n = 23) and in-patients (n = 15), moderate-high MICs of beta-lactams, except cefepime and carbapenems, were determined. All isolates were resistant to trimethoprim sulphamethoxazole. Multidrug resistance was detected in 58% of the isolates. The genes bla(CMY-2) and bla(CMY-7) were linked to ISEcp1 upstream in 32 cases and in one case, respectively, and bla(DHA-1) was linked to qacEDelta1sul1 upstream and downstream in one case. Twenty isolates were of phylogenetic groups B2 or D. Thirty-three XbaI-PFGE types, including three clusters, were observed. Twenty five sequence types (ST) were identified, of which ST complexes (STC) 38 (n = 7), STC 448 (n = 5) and ST131 (n = 4) were dominant. Plasmid profiling revealed 1-4 plasmids (50-250 kb) per isolate and 11 different replicons in 37/38 isolates; bla(CMY-2) was carried on transferable multiple-replicon plasmids, predominantly of Inc groups I1 (n = 12), FII (n = 10) and A/C (n = 7). Chromosomal integration was observed for bla(CMY-2) in ten strains. CMY-2 is the dominant PABL type in Norway and is associated with ISEcp1 and transferable, multiple-replicon IncI1, IncA/C, or IncFII plasmids in nationwide strains of STC 448, STC 38 and ST131. PMID- 19548923 TI - Utility of C-reactive protein in assessing the disease severity and complications of community-acquired pneumonia. AB - Previous studies on the usefulness of C-reactive protein (CRP) in patients with community-acquired pneumonia (CAP) have yielded somewhat inconsistent results. Our aim was to assess the value of CRP in estimating the severity and complications of CAP. CRP levels during the first 5 days of hospitalization were measured in 384 adult patients with CAP, and the data were evaluated using comprehensive statistical analyses. Significantly higher CRP levels on admission were detected in Pneumonia Severity Index (PSI) classes III-V than in classes I and II (p <0.001). An increment of 50 mg/L CRP on admission was associated with a 1.22-fold odds for a patient to be in PSI classes III-V as compared with classes I and II (OR 1.22, 95% CI 1.11-1.34; p <0.001). CRP levels were significantly higher in bacteraemic pneumonia than in non-bacteraemic pneumonia (p <0.001). An increment of 50 mg/L CRP was associated with a 1.67-fold odds for a patient to be bacteraemic (OR 1.67, 95% CI 1.46-1.92; p <0.001). CRP levels >100 mg/L on day 4 after the admission were significantly associated with complications (p <0.01). There was a trend for an association between the level of CRP on admission and the time to reach clinical stability (p <0.01). In conclusion, CRP may be valuable for revealing the development of complications in CAP. It may also be useful to assess the disease severity, thus being complementary to the assessment of the PSI. In our patients, high CRP levels were associated with a failure to reach clinical stability. PMID- 19548924 TI - In vitro activity of secnidazole against Atopobium vaginae, an anaerobic pathogen involved in bacterial vaginosis. AB - Bacterial vaginosis is a polymicrobial syndrome. The most important marker for bacterial vaginosis is the presence of Gardnerella vaginalis and Atopobium vaginae. In this study, the in vitro susceptibilities to metronidazole and secnidazole of 16 strains of A. vaginae were tested with the agar dilution method. We observed an MIC range for metronidazole of 4-64 mg/L (MIC(50), 8 mg/L; MIC(90), 32 mg/L) and an MIC range for secnidazole of 4-128 mg/L (MIC(50), 16 mg/L; MIC(90), 64 mg/L). According to these findings, we can conclude that the activity of secnidazole is similar to that of metronidazole. PMID- 19548925 TI - Comparison of multiple typing methods for Aspergillus fumigatus. AB - As part of studies on the spread of infections, risk factors and prevention, several typing methods were developed to investigate the epidemiology of Aspergillus fumigatus. In the present study, 52 clinical isolates of A. fumigatus from 12 airway specimens from patients with invasive aspergillosis (hospitalized in three different centres) were characterized by short tandem repeat (STR) typing and multilocus sequence typing (MLST). These isolates were previously typed by random amplified polymorphic DNA (RAPD), sequence-specific DNA polymorphism (SSDP), microsatellite polymorphism (MSP) and multilocus enzyme electrophoresis (MLEE). STR typing identified 30 genotypes and, for most patients, all isolates were grouped in one cluster of the unweighted pair group method with arithmetic mean dendrogram. Using MLST, 16 genotypes were identified among 50 isolates, while two isolates appeared untypeable. RAPD, MSP, SSDP and MLEE allowed identification of eight, 14, nine and eight genotypes, respectively. Combining the results of these methods led to the delineation of 25 genotypes and a similar clustering pattern as with STR typing. In general, STR typing led to similar results to the previous combination of RAPD, SSDP, MSP and MLEE, but had a higher resolution, whereas MLST was less discriminatory and resulted in a totally different clustering pattern. Therefore, this study suggests the use of STR typing for research concerning the local epidemiology of A. fumigatus, which requires a high discriminatory power. PMID- 19548926 TI - Clinical response to ertapenem in severe community-acquired pneumonia: a retrospective series in an elderly population. AB - To evaluate in routine hospital practice the clinical response to ertapenem in comparison with other parenteral antibiotics in the treatment of community acquired pneumonia (CAP), clinical records from patients with severe CAP treated with ertapenem from July 2002 to June 2006 in seven Spanish hospitals were retrospectively reviewed. Patients were classified according to the Pneumonia Severity Index (PSI). Each ertapenem-treated patient was matched with two patients in the same hospital treated with other antibiotics, according to age (difference 76 years). Comorbidities were present in 193 patients (95.5%). No differences were found in median hospital stay (7 days for ertapenem vs. 10 days for comparators, p 0.066). A slightly higher clinical response rate was obtained for ertapenem vs. comparators (88.7% vs. 77.1%; p 0.0465; OR 2.25; 95% CI 0.99-5.12), with significant differences in clinical response in patients coming from nursing homes (95.8% ertapenem vs. 63.8% comparators; p 0.0034) but not in non-institutionalized patients (85.4% ertapenem vs. 84.5% comparators; p 0.929). The higher clinical response to ertapenem vs. comparators in severe CAP was due to its significantly higher efficacy in healthcare-associated CAP in patients coming from nursing homes. PMID- 19548927 TI - Clinical characteristics of Haemophilus influenzae meningitis in Denmark in the post-vaccination era. AB - The introduction of Haemophilus influenzae type b (Hib) vaccine into the Danish childhood vaccination programme in 1993 may have influenced the epidemiology of H. influenzae meningitis (i.e. increasing frequency of other non-vaccine types; presentation in other age groups). Based on nationwide registration, clinical information and laboratory findings were collected from all 65 confirmed cases of H. influenzae meningitis during the period 1994-2005. Twenty-nine patients (45%) were <13 years old [median 15 months (range 0-147)], and 36 patients (55%) were >24 years old [median 62 years (range 25-96)]. Hib accounted for 31% (20/65) of the cases, and significantly more children were infected with Hib compared with adults [53% (16/29) vs. 11% (4/36), respectively, p 0.0003]. Overall, 38% of cases had an otogenic focus and this was thus the most frequent primary focus of infection. Among children infected with Hib, two cases (13%) were identified as true vaccine failures. Six patients (9%) died; one premature infant infected with serotype f and five adults (age 83-96 years) with non-typeable H. influenzae. Hearing loss was reported in 16% of the surviving children and in 10% of the surviving adults. The presence of a lung focus was an independent prognostic factor for an unfavourable outcome (p 0.03). In conclusion, meningitis caused by Hib has been infrequent in Denmark after introduction of the Hib vaccine in the childhood vaccination programme, and no increase in meningitis cases due to non-b type H. influenzae has been observed. Cases with H. influenzae meningitis frequently had an otogenic focus, with low risk of an unfavourable outcome. PMID- 19548928 TI - Epidemiological and clinical features of norovirus gastroenteritis in outbreaks: a population-based study. AB - Noroviruses are the most frequent cause of acute gastroenteritis in the community. In Catalonia, it is not clear how this type of viral gastroenteritis is evolving, and the objective of this prospective population-based study was to describe the incidence and epidemiological and clinical features of outbreaks of acute gastroenteritis due to norovirus in Catalonia between October 2004 and October 2005. Incidence rates were calculated using the estimated population of Catalonia in 2005. For each outbreak, the mode of transmission, the number of persons affected, demographic variables, clinical presentation, the date and time of onset of symptoms and the duration of symptoms, physician visits and hospitalizations were collected. Sixty viral outbreaks affecting 1791 people were identified, with no distinct seasonality. The mean number of outbreaks per month was 4.6. The global incidence was 24.6 per 100 000 person-years. The incidence was higher in women (25.7 per 100 000 person-years) and in the 5-11 years (52.4 per 100 000 person-years) and > or =65 years (42.4 per 100 000 person-years) age groups. The prevalence of vomiting, abdominal pain and general malaise was higher in children and adolescents, whereas the prevalence of diarrhoea and myalgia was higher in adults. These results suggest that norovirus infection has an important public health impact in Catalonia and that prevention strategies should be designed and implemented. PMID- 19548929 TI - Limitations of previously published systematic reviews evaluating the outcome of endodontic treatment. AB - The aim of this work was to identify the limitations of previously published systematic reviews evaluating the outcome of root canal treatment. Traditionally, periapical radiography has been used to assess the outcome of root canal treatment with the absence of a periapical radiolucency being considered a confirmation of a healthy periapex. However, a high percentage of cases confirmed as healthy by radiographs revealed apical periodontitis on cone beam computed tomography (CBCT) and by histology. In teeth, where reduced size of the existing radiolucency was diagnosed by radiographs and considered to represent periapical healing, enlargement of the lesion was frequently confirmed by CBCT. In clinical studies, two additional factors may have further contributed to the overestimation of successful outcomes after root canal treatment: (i) extractions and re-treatments were rarely recorded as failures; and (ii) the recall rate was often lower than 50%. The periapical index (PAI), frequently used for determination of success, was based on radiographic and histological findings in the periapical region of maxillary incisors. The validity of using PAI for all tooth positions might be questionable, as the thickness of the cortical bone and the position of the root tip in relation with the cortex vary with tooth position. In conclusion, the serious limitations of longitudinal clinical studies restrict the correct interpretation of root canal treatment outcomes. Systematic reviews reporting the success rates of root canal treatment without referring to these limitations may mislead readers. The outcomes of root canal treatment should be re-evaluated in long-term longitudinal studies using CBCT and stricter evaluation criteria. PMID- 19548930 TI - Microflora in teeth associated with apical periodontitis: a methodological observational study comparing two protocols and three microscopy techniques. AB - AIM: The aim of this study was to compare two protocols to examine bacterial colonization in teeth associated with chronic apical periodontitis with acute episodes (ap), using light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). METHODOLOGY: Nine root samples (seven teeth) were processed using either Eastman Dental Institute (EDI) (n = 4 teeth/4 roots) or Zurich (n = 3 teeth/5 roots) protocols. The roots were sectioned longitudinally; one root portion was viewed with SEM, descriptively dividing its length into apical, middle and coronal; semi-thin and ultra-thin transverse sections were viewed under LM and TEM from each third of the other root portion. Each root was therefore examined using all microscopy techniques. Observations of bacterial presence, description and distribution within the root canal lumen and root dentine were systematically recorded using pre-determined criteria. RESULTS: The Zurich technique gave a more predictable division of the root, but the surface was slightly smeared and demineralization was incomplete. The Eastman Dental Institute (EDI) approach appeared to provide better ultrastructural detail. Bacteria were detected in eight of the nine roots. Bacterial biofilms were commonly seen adhering to the root canal surface, containing various cellular morphotypes: rods, cocci, filaments and spirochaetes. Bacteria were more evident apically than coronally, associated with the canal wall but were more commonly evident coronally than apically within the dentinal tubules. Polymorphs (PMNs) were found in all the root thirds, especially apically, often numerous and walling off the bacterial biofilm from the remaining canal lumen. CONCLUSIONS: Both protocols had merits and de-merits. The combination of microscopy techniques offered complementary views of intra radicular bacterial colonization. The perception of confinement of the host/microbial interface at the apical foramen is not entirely correct; PMNs may be found even in the coronal third of root canals containing necrotic pulp tissue. PMID- 19548931 TI - A comparative study of image quality and radiation exposure for dental radiographs produced using a charge-coupled device and a phosphor plate system. AB - AIM: To investigate the quality of periapical radiographic images produced by two digital dental radiography systems, a charge-coupled device (CCD) and a photostimulable phosphor (PSP) image plate system, and to examine the overall radiation exposure when using these systems in a clinical setting. METHODOLOGY: Patients were randomly allocated to both systems and the resultant radiographs rated for quality. The expected radiation exposure for an investigation was calculated. RESULTS: Overall, 98 images were acquired using the CCD system and 108 with the PSP system. The PSP system produced significantly higher quality (P < 0.001) periapical images compared with the CCD system. The CCD system required significantly more (P < 0.001) repeat exposures to obtain a diagnostic image than the PSP system but at a lower expected radiation exposure. CONCLUSIONS: The image quality was superior using the phosphor plate system. Although more repeat radiographs were required using the CCD system, the images were produced with a lower expected radiation exposure. PMID- 19548932 TI - Complicated crown fractures - management and treatment options. AB - AIM: Complicated crown fractures involve enamel, dentine and the pulp. The incidence of complicated crown fractures ranges from 2% to 13% of all dental injuries and the most commonly involved tooth is the maxillary central incisor. Various treatment modalities are available depending upon the clinical, physiological and radiographic status of the involved tooth. The aim of this article was to discuss techniques for the management of complicated crown fractures. SUMMARY: Management of complicated crown fractures depends upon several factors with the result that various clinical modalities have been suggested. Seven case reports of complicated crown fractures are presented and risks/benefits of the treatment plans are discussed. KEY LEARNING POINTS: Proper diagnosis, treatment planning and follow-up care are important factors in the prognosis of complicated crown fractures. Thorough understanding of the available treatment modalities and their specific indications is critical. The risks and benefits of each treatment option should be carefully evaluated during the treatment planning process. PMID- 19548933 TI - The evaluation of displacement resistance of glass FRC posts to root dentine using a thin slice push-out test. AB - AIM: To investigate and compare the displacement resistance of glass fibre reinforced composite (FRC) posts to root dentine after luting with different adhesive systems. METHODOLOGY: A total of 32 noncarious extracted human mandibular premolars were prepared for post-cementation using the FRC Postec system (Ivoclar Vivadent, Schaan, Liechtenstein) and divided into four groups (n = 8). The posts in each group were luted with one or other of the following materials. Group 1: Variolink II/Excite DSC (etch-and-rinse, dual-curing), group 2: Clearfil Esthetic Cement/ED Primer II (self-etch, dual-curing), group 3: Multilink/Multilink Primer (self-etch, chemical-curing) and group 4: Multilink Sprint (self-adhesive, dual-curing). Specimens were sectioned to obtain slices with the post in the centre and with the root dentine overlaid by the autopolymerizing acrylic resin on each side. The displacement resistance was measured using a Universal Testing Machine at a crosshead speed of 0.5 mm min( 1). The displacement resistance of the specimens were calculated and expressed in MPa. Data were analysed with one-way ANOVA and post hoc Tukey's test (P < 0.05). RESULTS: Mean (SD) values of displacement resistance data in MPa are as follows: group 1, 12.08 (2.13); group 2, 12.39 (2); group 3, 11.3 (1.23); group 4, 14.29 (1.84). There were statistically significant differences amongst the groups (P = 0.021). A statistically significant difference was observed for the displacement resistance values between groups 3 and 4 (P = 0.015), that is between Multilink/Multilink Primer and Multilink Sprint. CONCLUSIONS: Glass FRC posts luted with self-adhesive luting system exhibited higher displacement resistance than when luted with chemical-curing self-etch luting system. PMID- 19548934 TI - A comparative study of physicochemical properties of AH Plus, Epiphany, and Epiphany SE root canal sealers. AB - AIM: To assess the physicochemical properties and the surface morphology of AH Plus, Epiphany, and Epiphany SE root canal sealers. METHODOLOGY: Five samples of each material were employed for each test according to ANSI/ADA specification 57. The samples were assigned to four groups: (i) AH Plus; (ii) Epiphany; (iii) Epiphany + Thinning Resin; (iv) Epiphany SE. The distilled water used during the solubility test was submitted to spectrometry to verify the release of calcium ions. The morphologies of the external surface and the cross-section of the samples were analysed by means of a scanning electron microscope (SEM). Statistical analysis was performed by using One-Way ANOVA and post hoc Tukey Kramer tests with the null hypothesis set as 5%. RESULTS: Setting time, flow and radiopacity results were in accordance with ANSI/ADA requirements whereas the dimensional change of all sealers and solubility of Epiphany did not fulfil ANSI/ADA protocols. AH Plus and Epiphany SE were similar in terms of flow, radiopacity, solubility and dimensional change. The spectrometry test revealed significant calcium ion release from Epiphany with and without the thinning resin. SEM analysis revealed essentially a homogeneous surface with compact layer and some rough areas. CONCLUSIONS: Setting time, flow, and radiopacity tests conformed to ANSI/ADA standardization. The dimensional change in all groups and the solubility of Epiphany were greater than values considered acceptable, with higher amounts of calcium ion release. Epiphany SE revealed more organized, compacted, and homogeneous polymers in a reduced resin matrix when compared with the other groups. PMID- 19548935 TI - The prevalence of anterior teeth with dens invaginatus in the western Mediterranean region of Turkey. AB - AIM: To assess the prevalence and type of dens invaginatus in anterior teeth of a selected population and to report associated dental complications. METHODOLOGY: The records of patients examined in the Department of Paedodontics, Faculty of Dentistry, Suleyman Demirel University, Isparta, Turkey between 1999 and 2006 were screened and a total of 2477 patients who had complete records with satisfactory radiographs were selected. The type of dens invaginatus and the presence of apical pathosis was determined from radiographs. Other dental abnormalities, syndromes and systemic diseases were noted. Variations in crown shape were also recorded. RESULTS: Dens invaginatus was detected in 300 out of 2477 patients, with a prevalence of 12%; 82% of affected patients had dens invaginatus bilaterally. Maxillary lateral incisors were the most affected teeth. The majority of the teeth had normal crown morphology (95%). The most commonly seen type of dens invaginatus was type I (94%). Overall 33% of the patients with type III dens invaginatus and 4% of the patients with type II dens invaginatus had apical pathosis. No associations with other systemic diseases and syndromes and some limited association with hypodontia and dens evaginatus were detected. CONCLUSION: A careful oral examination, radiographs, a suitable treatment plan and follow-up programme are crucial for early diagnosis and treatment for teeth with dens invaginatus. PMID- 19548936 TI - Endodontics or implants? A review of decisive criteria and guidelines for single tooth restorations and full arch reconstructions. AB - This review describes practical criteria and a systematic process to aid the treatment planning decision of whether to preserve teeth by root canal treatment (RCT) or extract and provide an implant. Recommendations presented are based on best available evidence from the literature and the expert views of specialists in endodontics and restorative dentistry, including dental implantology. A MEDLINE search was conducted using the terms 'root canal therapy', 'dental implants', 'decision making', 'treatment planning', 'outcome' and 'human', and supplemented by hand-searching. When evaluating the outcome of root canal treatment, an observation period of 4-5 years is required for complete healing of periapical lesions. Dental implants, however, present a de novo situation and a functional period of at least 5 years is often required before peri-implant diseases are established and detected. Good long-term success rates and greater flexibility in clinical management indicate that RCT or retreatment should be performed first in most instances unless the tooth is judged to be unrestorable. When deciding if a compromised tooth of questionable prognosis should be maintained or replaced by an implant, both local, site-specific and more general patient-related factors should be considered. Following systematic evaluation and consideration of the best treatment option in a particular case, a treatment recommendation may then be given in favour or against tooth retention. Whilst single risks are possibly accepted for single tooth restorations, teeth with questionable prognosis and multiple pre-treatment requirements are better not included as abutments in fixed dental prostheses to reduce the risk to survival of the entire restoration. PMID- 19548937 TI - Confocal laser scanning microscopy-guided surgery for neurofibroma. AB - The neurofibromatoses comprise at least two separate genetic disorders with variable clinical features and an unpredictable course. The most common type, neurofibromatosis 1, is characterized by > or = 6 cafe-au-lait spots and the occurrence of neurofibromas, which may present as cutaneous, subcutaneous or plexiform lesions. Normally, excision of neurofibromas is only indicated in the presence of neurological symptoms, suspicion of malignancy or for exceptional cosmetic reasons. For a good functional and aesthetic result with the least danger of recurrence, the surgeon's goal is to excise as much tissue as necessary and as little tissue as possible. One of the main issues during the surgical procedure is to distinguish between neurofibroma and surrounding tissue. We report for the first time the use of confocal laser scanning microscopy to differentiate between neurofibroma and healthy skin. PMID- 19548938 TI - Immunohistochemical clues to the diagnosis of Mondor's disease of the penis. AB - Mondor's disease of the penis is usually a benign, self-limiting process of acute onset. To date, there have been few studies about its diagnosis and treatment. It is difficult to differentiate MD of the penis from sclerosing lymphangitis of the penis, because the symptoms, epidemiology and aetiology are similar, and because veins are histologically very similar to lymphatics. We report a case of MD in which immunochemical markers such as CD31 and D240 allowed us to specifically discriminate between the small vein and lymphatic vessels. This is important not only for the recognition of MD at any site, but for many other diseases in which veins or lymphatic vessels are affected. PMID- 19548939 TI - Severe hypersensitivity reaction to minocycline in association with lymphomatoid papulosis. PMID- 19548940 TI - A case of acquired multiple plaque-like glomangiomyoma. PMID- 19548941 TI - Preauricular eczema: a sign of cellular phone dermatitis. PMID- 19548942 TI - Radiofrequency-selective nerve ablation for glabellar frown lines in East Asian patients. PMID- 19548943 TI - Stevens-Johnson syndrome due to prophylactic fluconazole in two patients with liver failure. PMID- 19548944 TI - Cutaneous vasculitis in a patient with acute brucellosis. PMID- 19548946 TI - Cytokine profile during the clinical course of toxic shock syndrome. AB - Toxic shock syndrome (TSS) is an acute febrile disease with multiple organ involvement caused by massive and rapid release of cytokines induced by staphylococcal exotoxins. However, the precise cytokine profile is still undefined in clinical cases. We measured serum cytokine concentrations in a patient who developed TSS after a caesarean section. Measurements were taken on admission and several times during the course of the disease. Methicillin resistant Staphylococcus aureus producing TSS toxin-1 and staphylococcal enterotoxin C was detected in the lochia and venous blood. Serum interleukin (IL) 6 level was markedly increased on admission, and IL-10, tumour necrosis factor alpha, and interferon-gamma levels were also raised. These cytokine levels rapidly returned to normal levels. In contrast, IL-1beta and IL-2 were below the analytical sensitivity threshold throughout the course. Our data and other previous case reports indicate that a marked increase in IL-6 concentration could be a clinical marker of TSS onset. PMID- 19548945 TI - Aquagenic palmar wrinkling as a presenting feature of cystic fibrosis gene dysfunction. AB - Aquagenic palmar wrinkling (APW) is characterized by the rapid and transient oedematous wrinkling of the palms after brief immersion in water. APW has been associated with cystic fibrosis (CF). Since the discovery of the CF gene, the clinical spectrum of CF has broadened from classic severe CF to include milder 'atypical CF' and 'CF-related disorders'. We report an unusual case in which APW occurred in a patient with no lung disease, and in whom investigations showed evidence of CF gene dysfunction. APW may be a presenting feature of a CF-related disorder and should prompt investigation of CF gene dysfunction. PMID- 19548947 TI - Jacquet erosive diaper dermatitis: a therapeutic challenge. PMID- 19548948 TI - Unrecognized radiation-induced localized scleroderma: a cause of postoperative wound-healing disorder. PMID- 19548949 TI - Polymorphic eruption of pregnancy in a photodistribution: a potentially new association? PMID- 19548950 TI - Thalidomide-induced polyneuropathy: friend or foe for relief of itch? PMID- 19548951 TI - Anetodermic pilomatricoma in a patient with tuberous sclerosis. PMID- 19548952 TI - Leptin secretion rate increases with higher CAG repeat number in Huntington's disease patients. AB - BACKGROUND: Huntington's disease (HD) is a hereditary neurodegenerative disorder caused by an increased number of CAG repeats in the huntingtin gene. A hallmark of HD is unintended weight loss, the cause of which is unknown. OBJECTIVE: To perform a detailed analysis of adipose tissue function in HD patients as abnormal fat tissue function could contribute to the weight loss. DESIGN, SETTING AND PARTICIPANTS: In a clinical research laboratory, 24-h plasma concentrations of leptin, adiponectin and resistin were studied in nine early-stage, medication free HD patients and nine age-, gender- and body mass index (BMI)-matched controls. MEASUREMENTS: Leptin was measured every 20 min whereas adiponectin and resistin were measured hourly. Autodeconvolution and cosinor regression were applied to quantify secretion characteristics of leptin and diurnal variations in leptin, adiponectin and resistin levels. RESULTS: Plasma levels and diurnal rhythmicity of leptin, adiponectin and resistin were not significantly different between HD patients and controls. However, although leptin production increased with higher BMI and fat mass in controls, no such relation was present in HD patients. Moreover, when corrected for fat mass, mean plasma leptin concentration as well as basal, pulsatile and total secretion rates increased with the size of the CAG repeat mutation (r = +0.72 to r = +0.80; all P < 0.05). Both higher pulsatile leptin secretion and higher mean adiponectin levels were associated with a greater degree of motor and functional impairment in HD patients. CONCLUSIONS: CAG-repeat size-dependent interference of the HD mutation with adipose tissue function may contribute to weight loss in HD patients. PMID- 19548954 TI - Reflections about "burn-out". PMID- 19548955 TI - Microarray analysis of placental tissue in intrauterine growth restriction. AB - OBJECTIVE: Besides foetal or maternal disorders, placental dysfunction is a major cause of intrauterine growth restriction (IUGR). Although numerous macro- and histopathological changes have been described, little is known about the precise aetiology and the contribution of foetal/placental genes in this disorder. DESIGN: Placental tissues of 20 IUGR and control neonates were analysed by microarray technique. Four of the regulated genes with possible relevance in the pathogenesis of IUGR and its consequences were further studied in placentas of 27 IUGR and 35 control newborns. RESULTS: Elevated gene expression of leptin, corticotrophin-releasing hormone (CRH), and IGF-binding protein-1 (IGFBP-1) in IUGR placentas could be confirmed in the larger group by real-time PCR, whereas prolactin showed no significant difference. Accordingly, protein expression of leptin and IGFBP-1 depicted by Western blot was elevated in IUGR, prolactin was not different. Birthweight standard deviation score (SDS) correlated negatively to leptin, IGFBP-1, and CRH, whereas placental weight correlated only to IGFBP-1. Leptin correlated negatively to gestational age of IUGR patients and positively to placental score, a marker of severity of impaired foeto-placental circulation. CONCLUSIONS: As confirmed in a large group of IUGR and control samples, the up regulated factors leptin, IGFBP-1, and CRH may serve as candidate genes for the prediction of subsequent metabolic consequences in IUGR newborns. These three factors may not only influence growth of the foetus, but might also interact with programming of its metabolic functions, which has to be determined in an ongoing study. PMID- 19548956 TI - In vivo comparison of force development with various materials of implant supported prostheses. AB - The purpose of this study was to measure axial loading, generating bending moments on fibre reinforced composite (FRC) implant prostheses using strain gauged customized abutment in vivo. Bending moments of conventional implant prosthetic material were also measured and the data were compared with those for FRC. Three unit fixed dental prostheses were made for two dental implant fixtures, which had been functioning properly for more than one year using Tescera ATL, porcelain fused to metal and gold as occlusal material. Three patients participated in this study; two patients had two implants on one side of the mandible and one had two implants on both sides of the mandible. Five sets of fixed dental prostheses were fabricated for each material and these were cemented with Temp-bond on strain-gauged customized abutments, which were screwed into the underlying implant fixtures. Axial loadings and bending moments were measured when a patient bit the experimental fixed dental prosthesis. ANOVA and the Tukey HSD test (alpha = 0.05) were used for statistical analysis. There were no significant differences in normalized bending moments among the three different implant prosthetic materials. Within the limitations of this study, Tescera ATL generated bending moments similar to conventional implant prosthetic materials such as gold and porcelain. PMID- 19548957 TI - Relation of dietary preference to bite force and occlusal contact area in Japanese children. AB - The purpose of this study was to examine the relationship of dietary preference to bite force and occlusal contact area in Japanese elementary school children. A total of 348 children, aged 7-12 years, from two public elementary schools located in Okayama Prefecture, Japan, participated in the study. Clinical examination included decayed, missing and filled teeth (dmft and DMFT), and total numbers of deciduous and permanent teeth. Bite force and occlusal contact area were measured using a pressure-detecting sheet. Dietary preference was assessed using a questionnaire in which the answers were given in like/dislike form. Mann Whitney U-test and multiple logistic regression analysis were applied to analyse the data. In multiple logistic regression analysis after adjustment for age, gender and total number of teeth present, children who liked cabbage and celery showed significantly higher bite force (P = 0.05 and P < 0.01, respectively) than those who disliked these. Children who liked cabbage and celery also showed higher occlusal contact area (P < 0.05 and P < 0.01, respectively) than those who disliked these. The Japanese elementary school children who liked hard foods such as cabbage and celery showed higher bite force and higher occlusal contact area than those who disliked these foods. A positive attitude towards harder food items might contribute to healthy development of the masticatory apparatus. PMID- 19548958 TI - Electromyographic standardized indices in healthy Brazilian young adults and data reproducibility. AB - The determination of normal parameters is an important procedure in the evaluation of the stomatognathic system. We used the surface electromyography standardization protocol described by Ferrario et al. (J Oral Rehabil. 2000;27:33 40, 2006;33:341) to determine reference values of the electromyographic standardized indices for the assessment of muscular symmetry (left and right side, percentage overlapping coefficient, POC), potential lateral displacing components (unbalanced contractile activities of contralateral masseter and temporalis muscles, TC), relative activity (most prevalent pair of masticatory muscles, ATTIV) and total activity (integrated areas of the electromyographic potentials over time, IMPACT) in healthy Brazilian young adults, and the relevant data reproducibility. Electromyography of the right and left masseter and temporalis muscles was performed during maximum teeth clenching in 20 healthy subjects (10 women and 10 men, mean age 23 years, s.d. 3), free from periodontal problems, temporomandibular disorders, oro-facial myofunctional disorder, and with full permanent dentition (28 teeth at least). Data reproducibility was computed for 75% of the sample. The values obtained were POC Temporal (88.11 +/- 1.45%), POC masseter (87.11 +/- 1.60%), TC (8.79 +/- 1.20%), ATTIV (-0.33 +/- 9.65%) and IMPACT (110.40 +/- 23.69 microV/microV.s %). There were no statistical differences between test and retest values (P > 0.05). The Technical Errors of Measurement (TEM) for 50% of subjects assessed during the same session were 1.5, 1.39, 1.06, 3.83 and 10.04. For 25% of the subjects assessed after a 6-month interval, the TEM were 0.80, 1.03, 0.73, 12.70 and 19.10. For all indices, there was good reproducibility. These electromyographic indices could be used in the assessment of patients with stomatognathic dysfunction. PMID- 19548959 TI - Adherence to topical dermatological therapy: lessons from oral drug treatment. AB - Patients are remarkably nonadherent to medical treatment regimens across all diseases and classes of therapy, and it has been estimated that nonadherence to drug treatment is responsible for as many as 10% of all hospital admissions. Nonadherence to treatment also has significant negative effects on treatment outcomes across a wide range of diseases. Patient-related factors such as age, ethnicity, literacy (including health literacy), health beliefs, and socioeconomic conditions have been shown to influence adherence to oral therapy. Medication-related factors, such as regimen complexity and duration of treatment, also impact on adherence. Variables that significantly influence adherence to oral drugs have similar effects on adherence to topical therapy. Both educational and psychological interventions along with simplification of dosing regimens can significantly improve adherence to oral therapy and limited evidence indicates that these approaches are also effective in patients receiving topical therapy. There is very little information about the effects of dosing regimens on adherence to topical medical therapy. The advent of new drug formulations that permit once-daily or single-dose drug application will, however, permit evaluation of different topical treatment regimens on adherence and treatment outcomes in patients with dermatological disease. PMID- 19548960 TI - Epidermal stem cells: practical perspectives and potential uses. AB - Throughout adult life, the epidermis and the hair follicle undergo a perpetual cycle of growth, regression and rest. Stem cells in the epidermis not only ensure the maintenance of epidermal homeostasis and hair regeneration, but also contribute to repair of the epidermis after injury. These stem cells lie within specific niches in the hair follicle and the epidermis. The availability of monoclonal antibodies that can be used on formalin-fixed paraffin-embedded tissue has greatly facilitated the use of this methodology as an adjunct to uncovering stem cell niches. In this review, we attempt to provide an overview of the potential markers available to identify and study stem cells in an effort to providing a better understanding of the pathogenesis of skin diseases including disorders of hair loss and malignancies. The potential uses of these markers in prognosis and in expanding the therapeutic options in several disorders will also be addressed. PMID- 19548961 TI - Analysis of current data on the use of methotrexate in the treatment of pemphigus and pemphigoid. AB - Methotrexate (MTX) is primarily used in the treatment of malignancies. It has also been used as an immunosuppressive agent in the treatment of pemphigus and pemphigoid. The objective of this study was to determine the role of MTX in the treatment of pemphigus and pemphigoid based on an analysis of the available literature. A retrospective analysis of the English language literature was conducted. The studies included in this analysis were required to fulfil the following inclusion criteria: English language; diagnosis based on histology and immunopathology; minimum of five patients in each series; and data for efficacy, spectrum of responses and follow-up. A total of 136 patients with pemphigus were reported in seven studies. One hundred and eleven of the 136 patients (82%) showed clinical improvement with MTX. A total of 79 patients with pemphigoid were reported in six studies. Overall, 74 of the 79 patients (94%) showed clinical improvement. Nausea and infection were the most common side-effects. Death due to MTX resulted in seven of 215 patients (3%). There is a lack of randomized controlled trials. In many studies in this review there was insufficient information on clinical follow-up post-therapy and on serological correlations. Analysis of the data suggests that MTX may be useful and effective in patients with pemphigus vulgaris, who are corticosteroid dependent or who develop significant complications in relation to corticosteroids. MTX is likely to be more beneficial in patients with pemphigoid, particularly in bullous pemphigoid, than in patients with pemphigus. Given the limitations of the available data, it appears that when there is a need for adjuvant therapy, MTX may be considered early in the management of moderate to moderately severe disease. PMID- 19548962 TI - Blood group genotyping: from patient to high-throughput donor screening. AB - Blood group antigens, present on the cell membrane of red blood cells and platelets, can be defined either serologically or predicted based on the genotypes of genes encoding for blood group antigens. At present, the molecular basis of many antigens of the 30 blood group systems and 17 human platelet antigens is known. In many laboratories, blood group genotyping assays are routinely used for diagnostics in cases where patient red cells cannot be used for serological typing due to the presence of auto-antibodies or after recent transfusions. In addition, DNA genotyping is used to support (un)-expected serological findings. Fetal genotyping is routinely performed when there is a risk of alloimmune-mediated red cell or platelet destruction. In case of patient blood group antigen typing, it is important that a genotyping result is quickly available to support the selection of donor blood, and high-throughput of the genotyping method is not a prerequisite. In addition, genotyping of blood donors will be extremely useful to obtain donor blood with rare phenotypes, for example lacking a high-frequency antigen, and to obtain a fully typed donor database to be used for a better matching between recipient and donor to prevent adverse transfusion reactions. Serological typing of large cohorts of donors is a labour intensive and expensive exercise and hampered by the lack of sufficient amounts of approved typing reagents for all blood group systems of interest. Currently, high-throughput genotyping based on DNA micro-arrays is a very feasible method to obtain a large pool of well-typed blood donors. Several systems for high throughput blood group genotyping are developed and will be discussed in this review. PMID- 19548963 TI - Prion removal effect of a specific affinity ligand introduced into the manufacturing process of the pharmaceutical quality solvent/detergent (S/D) treated plasma OctaplasLG. AB - BACKGROUND AND OBJECTIVES: A new chromatographic step for the selective binding of abnormal prion protein (PrP(Sc)) was developed, and optimization for PrP(Sc) capture was achieved by binding to an affinity ligand attached to synthetic resin particles. This step was implemented into the manufacturing process of the solvent/detergent (S/D)-treated biopharmaceutical quality plasma Octaplas to further improve the safety margin in terms of risk for variant Creutzfeldt-Jakob disease (vCJD) transmission. MATERIALS AND METHODS: Intermediates and Octaplas final container material, spiked with hamster brain-derived PrP(Sc)-containing fractions, were used for experiments to establish the feasibility of introducing this novel chromatography step. The binding capacity per millilitre of ligand gel was determined under the selected manufacturing conditions. In addition, the specificity of the ligand gel to bind PrP(Sc) from human sources was investigated. A validated Western blot test was used for the identification and quantification of PrP(Sc). RESULTS: A reduction factor of > or = 3.0 log(10) could be demonstrated by Western blotting, utilizing the relevant Octaplas matrix from manufacturing. In this particular cell-free plasma solution, the PrP(Sc) binding capacity of the selected gel was very high (> or = 6 log(10) ID(50)/ml, equivalent to roughly 10 log(10) ID(50)/column at manufacturing scale). The gel binds specifically PrP(Sc) from both animal (hamster and mouse) and human (sporadic and variant CJD) sources. CONCLUSION: This new single-use, disposable PrP(Sc)-harvesting gel ensures a very high capacity in terms of removing the pathogenic agent causing vCJD from the new generation OctaplasLG, in the event that prions can be found in plasma from donors incubating the disease and thereby contaminating the raw material plasma used for manufacturing. PMID- 19548964 TI - Respiratory patterns in panic disorder reviewed: a focus on biological challenge tests. AB - OBJECTIVE: To provide a systematic review of studies investigating respiration in PD and comments on relative inconsistencies. METHOD: A Medline search of controlled studies focusing on pCO(2), respiratory rate, tidal volume, and minute volume in PD patients was conducted for baseline/resting condition, challenge, and recovery phase. Respiratory variability and comparisons between panickers and non-panickers were also examined. RESULTS: Lower pCO(2) levels in PD subjects are a consistent finding during the baseline/resting condition, the challenge, and recovery phases. Tidal volume and minute volume are increased in PD subjects relative to controls during the baseline/resting condition. However, the most robust finding is a higher than normal respiratory variability, which appears to be a promising factor for the identification of respiratory etiopathological pathways in PD. CONCLUSION: Respiratory variability might be a candidate for a biological marker of PD: an abnormal breathing pattern as found in panic disorder (PD) patients compared with controls might indicate instability of the respiratory homeostasis. PMID- 19548965 TI - Body mass index and acute asthma severity among children presenting to the emergency department. AB - To determine the prevalence of obesity among children presenting to the emergency department (ED) with acute asthma, and to examine the relationship between body mass index (BMI) and acute asthma severity in the ED setting. We analyzed data from a multicenter prospective cohort study during 1997-1998; 44 ED in 17 US states and two Canadian provinces enrolled 672 patients, age 5-17, with acute asthma. BMI and Pulmonary Index were collected in the ED. We defined overweight and obesity using age, sex, and race-specific BMI values from national and international databases. The prevalence of obesity was significantly higher among ED patients with acute asthma as compared with children from the general population (23% vs. 9-15%; p < 0.001). Obese children with acute asthma did not differ from their non-obese counterparts, by demographic factors or chronic asthma severity (all p > 0.2). Initial Pulmonary Index was the same across underweight, intermediate, and obese groups (3.7 +/- 2.4, 3.8 +/- 2.2, 3.7 +/- 2.3; p = 0.70). Admission status also did not vary across groups (22%, 22% and 23%; p = 0.98). Stratifying the analysis by age group and sex did not change these results. The prevalence of obesity among children presenting to the ED with acute asthma was significantly higher compared with children from the general population. BMI was not associated with markers of chronic and acute asthma severity. The results of this study support a positive association between obesity and asthma, and suggest that asthma exacerbations among obese children are very similar to those experienced by other children. PMID- 19548967 TI - Sarcocystis sp. encephalomyelitis in a cat. AB - A 5-month-old male neutered domestic shorthair cat was evaluated for spinal pain, ataxia, and anisocoria. Neuroanatomic localization indicated diffuse or multifocal central nervous system disease. On cerebrospinal fluid analysis, neutrophilic pleocytosis and intracellular protozoal merozoites were observed. The merozoites were oval, 2-4 microm in width and 4-6 microm in length, and had linear arrays of nuclear material concentrated at one pole. Serum was positive for Sarcocystis sp. antibodies and negative for Toxoplasma gondii antibodies. The organism was determined to be either Sarcocystis neurona or Sarcocystis dasypi based on sequence analysis of the internal transcribed spacer 1 ribosomal RNA genomic region. Clinical disease resolved following treatment with 3 different protocols for protozoal infection. This case is the first to demonstrate the antemortem diagnosis and survival of a domestic cat with Sarcocystis sp. associated encephalomyelitis. Clinicians and cytopathologists should include Sarcocystis sp. as a differential for feline inflammatory central nervous system disease characterized by neutrophilic pleocytosis. PMID- 19548966 TI - Early wheezing phenotypes and cognitive development of 3-yr-olds. Community recruited birth cohort study. AB - The main purpose of the study was to answer the question whether young children without clinical diagnosis of asthma but experiencing early wheezing disorders and therefore being at high risk of developing asthma may have cognitive deficits. In the ongoing birth cohort study wheezing symptoms were recorded postpartum over two first years of age and subsequently cognitive status of children at the age of 3 yr was assessed with the Bayley Mental Development Index (MDI). In the statistical analysis a wide range of modifying and confounding factors (maternal education, gender of children, prenatal exposure to lead and environmental tobacco smoke (ETS) were considered to assess the independent effect of early wheezing phenotypes on cognitive development of children. The MDI score correlated inversely with the number of wheezing days recorded over 24 months (r = -0.13, p = 0.007), lead cord blood concentration (r = -0.12, p = 0.02), number of siblings (r = -0.17, p = 0.0006) and the number of cigarettes smoked daily by other household members at home over the pregnancy period (r = 0.18, p = 0.0002). While the children who experienced wheezing over the first year of age showed deficit of 2 MDI scores (beta coeff. = -2.31, 95%CI: -4.63 to 0.02), those with persistent wheezing had the score deficit of 4 points (beta coeff. = -4.41, 95%CI: -8.27 to -0.55). To our knowledge, it is the first report in the iterature showing that early wheezing is associated the cognitive deficit in a community-recruited very young children. Observed cognitive deficit in early wheezers may be caused by RSV infections or can be related to lower lung function attributed to persistent wheezing, which reducing oxygen supply would affect rapidly developing brain. PMID- 19548969 TI - Thrombelastography in dogs admitted to an intensive care unit. AB - BACKGROUND: Underlying conditions in dogs admitted to an intensive care unit (ICU) can cause hemostatic dysfunction. Thrombelastography (TEG) may be useful in detecting hemostatic alterations as compared with standard coagulation tests. OBJECTIVES: The purpose of this study was to compare TEG results and those of standard coagulation tests in identifying hemostatic dysfunction in dogs admitted to an ICU and to investigate associations among the variables measured. METHODS: Tissue factor-activated TEG analysis, d-dimer and fibrinogen concentrations, antithrombin (AT) activity, prothrombin time (PT), activated partial thromboplastin time (aPTT), and platelet count were measured using standard techniques on 27 dogs admitted to ICU with a disease known to be associated with hemostatic dysfunction and in 31 clinically healthy control dogs. Results were compared between groups using nonparametric tests and kappa analysis; principal component analysis (PCA) and Spearman rank correlation were used to measure associations among variables. RESULTS: Fourteen of 27 ICU dogs had abnormal TEG tracings, which were used to classify the dogs as hypercoagulable (n=11), hypocoagulable (n=3), or normocoagulable (n=13). Hypercoagulable dogs had significantly increased d-dimer (P=.03) and fibrinogen (P=.01) concentrations compared with normocoagulable dogs. In ICU dogs, positive associations were identified between maximum amplitude (MA), alpha-angle, fibrinogen concentration, and platelet count, and between PT, aPTT, and reaction time (R). Significant correlations were found between MA and fibrinogen (r(s)=.76, P<.001) and between reaction time (R) and PT (r(s)=.51, P=.003). CONCLUSIONS: TEG was useful in detecting hemostatic dysfunction in dogs in an ICU. Positive associations among variables may provide insight as to how overall coagulation status reflects alterations in clot strength and coagulation time. Dogs with TEG tracings indicative of hypercoagulability are likely in procoagulant states. Future studies of the incidence of thrombotic complications in dogs with hypercoagulable TEG tracings are warranted. PMID- 19548968 TI - Effects of in vitro hemodilution of canine blood on platelet function analysis using the PFA-100. AB - BACKGROUND: The platelet function analyzer (PFA)-100 is a point-of-care instrument previously evaluated in humans and dogs. In both species, artificially prolonged platelet closure time (CT) occurs with anemia. Reliability of the analyzer in dogs becomes a concern when the HCT is between 0.25 and 0.35 L/L. OBJECTIVE: The objective of this study was to further define the level of HCT at which CT is prolonged, using in vitro diluted canine blood. METHODS: Citrated whole blood samples were collected from 22 healthy dogs. Initial HCT was determined and autologous platelet-rich plasma was added to samples to achieve HCTs of 0.33, 0.30, and 0.27 L/L. CT was determined in duplicate on the PFA-100 using collagen/adenosine-5'-diphosphate cartridges. RESULTS: Compared with the initial CT in samples with HCT 0.39-0.54 L/L (CT mean+/-SD=57.8+/-5.75 seconds), significantly prolonged CTs were found in hemodiluted samples with HCT 0.33 L/L (61.1+/-4.64 seconds), 0.30 L/L (64.3+/-6.79 seconds), and 0.27 L/L (70.8+/-7.90 seconds) (P=0.029; repeated measures ANOVA). CONCLUSION: Although statistical differences were found, further studies are needed to determine the clinical significance of the mild prolongation in CT associated with mild anemia. Until then, dogs with HCTs slightly <0.35 L/L should be evaluated cautiously for platelet dysfunction using the PFA-100. PMID- 19548970 TI - Differential cell counts in canine cytocentrifuged bronchoalveolar lavage fluid: a study on reliable enumeration of each cell type. AB - BACKGROUND: Bronchoalveolar lavage (BAL) allows cell recovery from the lower respiratory tract; differential cell counts of BAL fluid gives important information in the assessment of various bronchial and pulmonary diseases. To the best of our knowledge no study has investigated the relation between the number of cells counted and the reproducibility of BAL fluid differential cell counts. OBJECTIVE: The purpose of this study was to investigate using statistical methods how many cells should be counted in cytocentrifuged BAL fluid preparations in order to obtain a reliable enumeration of each cell type. METHODS: BAL fluid samples from dogs with suspected bronchopulmonary disease were obtained during fiberoptic bronchoscopy with a standardized protocol. Differential cell counts were performed on May-Grunwald-Giemsa-stained cytocentrifuged preparations by 2 independent observers. Reproducibility for the enumeration of each cell type was expressed as the intraclass correlation coefficient. We considered a threshold level of >or=0.90 to be high and a threshold level of >or=0.85 to be adequate. RESULTS: Forty BAL fluid samples were included in the study. For neutrophils, alveolar macrophages, and eosinophils high reproducibility was reached by counting 200 cells; adequate reproducibility was reached for lymphocytes and bronchial epithelial cells by counting 500 cells. CONCLUSIONS: A 500-cell differential count is required for all types of cells to be quantified with adequate reproducibility in canine cytocentrifuged BAL fluid samples. PMID- 19548971 TI - Serum thyroxine concentrations in clinically healthy pet guinea pigs (Cavia porcellus). AB - BACKGROUND: Guinea pigs (Cavia porcellus) are often presented as patients in veterinary practice. Nevertheless, only limited information is available about endocrine diseases or thyroxine reference values for the species. OBJECTIVE: The aim of this study was to determine serum thyroxine concentrations in a well defined population of clinically healthy pet guinea pigs. METHODS: Between October 2007 and July 2008, serum samples were collected from 40 clinically healthy guinea pigs of different sexes, ages, and breeds that were presented to our clinic for a general health check or for castration. Pregnant females were excluded from the study. Thyroxine concentration was measured using a chemiluminescence test (Immulite 2000 Canine Total T4). RESULTS: Thyroxine concentrations ranged from 14.2 to 66.9 nmol/L (1.1-5.2 microg/dL) with a median value of 27.0 nmol/L (2.1 microg/dL). Females (n=16) had significantly (P=.039; Mann-Whitney U-test) lower thyroxine values than castrated males (n=8), whereas no differences were found between females and intact males (n=16) or between intact and castrated males. No significant correlation was found between thyroxine concentration and age. CONCLUSION: This is the first report of serum thyroxine reference values for a well-defined population of healthy pet guinea pigs as measured by a chemiluminescence assay. The results were higher than those previously reported for this species and emphasize the importance of using appropriate reference intervals for the diagnosis of hyperthyroidism. PMID- 19548972 TI - Design, optimization, and application of a conventional PCR assay with an internal control for detection of 'Candidatus Mycoplasma turicensis' 16S rDNA in domestic cats from Brazil. AB - BACKGROUND: 'Candidatus Mycoplasma turicensis' (CMtc) is a hemotrophic bacterial species that can, alone or in combination, induce anemia in cats. The diagnostic test of choice for hemoplasma infections is PCR. Conventional PCR assays have been developed for the detection of Mycoplasma haemofelis (Mhf) and 'Candidatus M. haemominutum' (CMhm) but not for CMtc. Although real-time PCR assays have been reported for all of the feline hemoplasmas, the expense of necessary instrumentation precludes its use in Brazil and many other countries. OBJECTIVES: The goals of this study were to develop and optimize a conventional PCR assay to diagnose CMtc using an internal control to detect false-negative results, and to evaluate the occurrence of CMtc infection in domestic cats from Brazil. METHODS: Species-specific primers were designed and a PCR assay was developed for the detection of CMtc 16S rDNA in cat blood. Sensitivity was determined by serial 10 fold dilutions of plasmid and DNA extracted from blood from an experimentally infected cat. EDTA blood samples from 373 cats were collected. DNA was extracted using a silica-based protocol and tested using the PCR assay. RESULTS: Primer concentration, annealing temperature, and MgCl(2) concentration were optimized in the presence and absence of the internal control. Two samples negative for the internal control were excluded. Of the remaining 371 samples (117 healthy and 254 unhealthy cats), 17 (4.6%) were positive for CMtc. CONCLUSION: These results demonstrate the utility of an optimized PCR assay to detect CMtc in feline blood samples. We also report for the first time the prevalence of CMtc infection in domestic cats in Brazil. PMID- 19548973 TI - Evaluation of a modified thrombelastography assay initiated with recombinant human tissue factor in clinically healthy horses. AB - BACKGROUND: Thrombelastography (TEG) is used to evaluate the viscoelastic properties of blood during clotting and provides a global assessment of hemostasis and clot lysis. TEG analysis initiated with recombinant human tissue factor (TF) has not been evaluated in clinically healthy horses. OBJECTIVES: The purpose of this study was to determine whether TEG results are affected by the time elapsed between sampling and analysis (storage time) of equine blood samples and to establish a preliminary equine reference interval for a modified TEG assay, using recombinant human TF to initiate coagulation. METHODS: Citrated blood samples were obtained from 20 clinically healthy adult horses. Thirteen samples were stored for 30, 60, and 120 minutes at room temperature before TEG analysis. Coagulation was initiated by adding 20 microL of CaCl(2) to 330 microL of blood and 10 microL of diluted recombinant TF for a final dilution of 1:3600. Reaction (R) and clotting (K) times, angle (alpha), and maximum amplitude (MA) were compared between time points. A preliminary reference interval (minimum maximum values) was determined using data from all 20 horses after 30 minutes of sample storage. RESULTS: There was a significant effect of storage time on R, K, and alpha but not MA. Reference intervals were: R, 3.65-6.4 minutes; K, 1.8-5.45 minutes; alpha, 33.4-66.2 degrees ; MA, 41.2-64.1 mm; lysis at 30 minutes post-MA (LY30), <2.75%; and lysis at 60 minutes post-MA (LY60), 1.55-9.5%. CONCLUSIONS: TEG can be performed on equine citrated blood samples using recombinant human TF to activate clot formation. TEG parameters were significantly affected by storage time, suggesting an incomplete inhibition of coagulation in citrated blood. PMID- 19548974 TI - Acute phase proteins in the diagnosis of bovine subclinical mastitis. AB - BACKGROUND: The California mastitis test (CMT) and somatic cell count (SCC) are commonly used for diagnosis of subclinical mastitis in cattle. Acute phase proteins (APPs), as alternative biomarkers of mastitis, may increase in concentration in the absence of macroscopic changes in the milk, or may precede the onset of clinical signs. OBJECTIVE: The aim of this study was to compare the accuracy of APPs measured in milk and in serum with bacterial culture for the diagnosis of bovine subclinical mastitis. METHODS: One hundred and seventy-five Holstein cows were randomly selected from 7 dairy farms. Quarter milk and serum samples were taken from all cows. Milk samples were analyzed using a CMT and SCC, and for haptoglobin (MHp) and amyloid A (MAA) concentrations, and were also submitted for bacterial culture. Serum samples obtained concurrently were analyzed for haptoglobin (SHp) and amyloid A (SAA). Two-sample Wilcoxon (Mann Whitney) test was used to compare SCC, MAA, MHp, SAA, and SHp concentrations between culture-positive and culture-negative animals. Receiver-operating characteristic analysis was used to assess the performance of each test using bacterial culture as the reference method. RESULTS: MAA concentration was the most accurate of the 5 tests, with a sensitivity of 90.6% and specificity of 98.3% at concentrations >16.4 mg/L. MAA and MHp had significantly larger areas under the curve than the respective serum proteins, SAA and SHp. CONCLUSIONS: The results suggest that measuring haptoglobin and amyloid A in milk is more accurate than serum analysis for the diagnosis of subclinical mastitis in Holstein cows. PMID- 19548975 TI - The immunocompromised district: a unifying concept for lymphoedematous, herpes infected and otherwise damaged sites. AB - Systemic immunodeficiency is known to facilitate the onset of opportunistic infections, tumours and immune disorders in any district of the body. There are clinical events, such as chronic lymphoedema, herpetic infections, vaccinations and heterogeneous physical injuries which can selectively damage and immunologically mark the cutaneous district they act upon. After the causing event has disappeared, the affected district may appear clinically normal, but its immune behaviour is often compromised forever. An immunocompromised district becomes a site which is particularly susceptible to subsequent outbreaks of opportunistic infections, tumours and immune disorders confined to the district itself. In this review, there is an ample case-report collection of opportunistic disorders (infectious, neoplastic, immune) which appeared in immunocompromised districts. The cases have been grouped according to the clinical settings responsible for the local immune imbalance: regional chronic lymphoedema; herpes infected sites, which feature the well-known Wolf's isotopic response; and otherwise damaged areas, comprising sites of vaccination, ionizing or UV radiation, thermal burns and traumas. Whatever the immunocompromising factor, a common denominator which facilitates the occurrence of tumours, infections and dysimmune reactions in an immunocompromised district may reside in locally hampered lymph drainage and/or locally altered neuromediator signalling. In fact, any obstacle to the normal trafficking of immunocompetent cells through lymphatic channels or any interference with the signals that the neuropeptides and neurotransmitters released by peripheral nerves send to cell membrane receptors of immunocompetent cells, can significantly alter the local immune response, thus paving the way for heterogeneous opportunistic disorders in the immunocompromised district. PMID- 19548976 TI - Outcome of granuloma annulare: shorter duration is associated with younger age and recent onset. PMID- 19548977 TI - Use of isotretinoin for photoaging. PMID- 19548978 TI - Unequally redundant RCD1 and SRO1 mediate stress and developmental responses and interact with transcription factors. AB - RADICAL-INDUCED CELL DEATH1 (RCD1) is an important regulator of stress and hormonal and developmental responses in Arabidopsis thaliana. Together with its closest homolog, SIMILAR TO RCD-ONE1 (SRO1), it is the only Arabidopsis protein containing the WWE domain, which is known to mediate protein-protein interactions in other organisms. Additionally, these two proteins contain the core catalytic region of poly-ADP-ribose transferases and a conserved C-terminal domain. Tissue and subcellular localization data indicate that RCD1 and SRO1 have partially overlapping functions in plant development. In contrast mutant data indicate that rcd1 has defects in plant development, whereas sro1 displays normal development. However, the rcd1 sro1 double mutant has severe growth defects, indicating that RCD1 and SRO1 exemplify an important genetic principle - unequal genetic redundancy. A large pair-wise interaction test against the REGIA transcription factor collection revealed that RCD1 interacts with a large number of transcription factors belonging to several protein families, such as AP2/ERF, NAC and basic helix-loop-helix (bHLH), and that SRO1 interacts with a smaller subset of these. Full genome array analysis indicated that in many cases targets of these transcription factors have altered expression in the rcd1 but not the sro1 mutant. Taken together RCD1 and SRO1 are required for proper plant development. PMID- 19548979 TI - The Arabidopsis TSPO-related protein is a stress and abscisic acid-regulated, endoplasmic reticulum-Golgi-localized membrane protein. AB - The Arabidopsis gene At2g47770 encodes a membrane-bound protein designated AtTSPO (Arabidopsis thaliana TSPO-related). AtTSPO is related to the bacterial outer membrane tryptophan-rich sensory protein (TspO) and the mammalian mitochondrial 18-kDa translocator protein (18 kDa TSPO), members of the group of TspO/MBR domain-containing membrane proteins. In this study we show that AtTSPO is mainly detected in dry seeds, but can be induced in vegetative tissues by osmotic or salt stress or abscisic acid (ABA) treatment, corroborating available transcriptome data. Using subcellular fractionation, immunocytochemistry and fluorescent protein tagging approaches we present evidence that AtTSPO is targeted to the secretory pathway in plants. Induced or constitutively expressed AtTSPO can be detected in the endoplasmic reticulum and the Golgi stacks of plant cells. AtTSPO tagged with fluorescent protein in transgenic plants (Arabidopsis and tobacco) was mainly detected in the Golgi stacks of leaf epidermal cells. Constitutive expression of AtTSPO resulted in increased sensitivity to NaCl, but not to osmotic stress, and in reduced greening of cultured Arabidopsis cells under light growing conditions. Transgenic Arabidopsis plants overexpressing AtTSPO were more sensitive to ABA-induced growth inhibition, indicating that constitutive expression of AtTSPO may enhance ABA sensitivity. AtTSPO is rapidly downregulated during seed imbibition, and the ABA-dependent induction in plant is transient. Downregulation of AtTSPO seems to be boosted by treatment with aminolevulinic acid. Taken together, these results suggest that AtTSPO is a highly regulated protein, induced by abiotic stress to modulate, at least in part, transient intracellular ABA-dependent stress perception and/or signalling. PMID- 19548980 TI - Microbial community analysis of rectal methanogens and sulfate reducing bacteria in two non-human primate species. AB - BACKGROUND: Methanogenesis by methanogenic Archaea and sulfate reduction by sulfate reducing bacteria (SRB) are the major hydrogenotrophic pathways in the human colon. Methanogenic status of mammals is suggested to be under evolutionary rather than dietary control. However, information is lacking regarding the dynamics of hydrogenotrophic microbial communities among different primate species. METHODS: Rectal swabs were collected from 10 sooty mangabeys (Cercocebus atys) and 10 baboons (Papio hamadryas). The diversity and abundance of methanogens and SRB were examined using PCR-denaturing gradient gel electrophoresis (DGGE) and real-time quantitative PCR (qPCR). RESULTS: The DGGE results revealed that intestinal Archaea and SRB communities differ between mangabeys and baboons. Phylogenetic analyses of Archaea DGGE bands revealed two distinct clusters with one representing a putative novel order of methanogenic Archaea. The qPCR detected a similar abundance of methanogens and SRB. CONCLUSIONS: Intestinal Archaea and SRB coexist in these primates, and the community patterns are host species-specific. PMID- 19548981 TI - Microarray kit analysis of cytokines in blood product units and segments. AB - BACKGROUND: Cytokine concentrations in transfused blood components are of interest for some clinical trials. It is not always possible to process samples of transfused components quickly after their administration. Additionally, it is not practical to sample material in an acceptable manner from many bags of components before transfusion, and after transfusion, the only representative remaining fluid of the component may be that in the "segment," because the bag may have been completely transfused. Multiplex array technology allows rapid simultaneous testing of multiple analytes in small-volume samples. This technology was used to measure white blood cell (WBC) cytokine levels in blood products to determine 1) whether concentrations in segments correlate with those in the main bag and, thus, whether segments could be used for estimation of the concentrations in the transfused component and 2) whether concentrations after sample storage at 4 degrees C for 24 hours do not differ from concentrations before storage, thus allowing for processing within 24 hours, rather than immediately after transfusion. STUDY DESIGN AND METHODS: WBC cytokines were measured in the supernatant from bags and segments of leukoreduced red blood cells (RBCs), nonleukoreduced whole blood, and leukoreduced plateletphereses using a human cytokine array kit (ProteoPlex, Novagen). RESULTS: Cytokine concentrations in RBCs and whole blood or plateletphereses stored at 4 degrees C did not differ between bag and segment samples (all p > 0.05). There was no evidence of systematic differences between segment and bag concentrations. Cytokine concentrations in samples from plateletphereses did not change within 24 hours storage at 4 degrees C. CONCLUSION: Samples from either bag or segment can be used to study cytokine concentrations in groups of blood products. Cytokine concentrations in plateletphereses appear to be stable for at least 24 hours of storage at 4 degrees C and, thus, samples stored with those conditions may be used to estimate the cytokine concentrations of the component at the time of transfusion. PMID- 19548983 TI - Importins and beyond: non-conventional nuclear transport mechanisms. AB - The movement of proteins between the cytoplasm and the nucleus conventionally involves the recognition of nuclear targeting signals by members of the importin (Imp) superfamily of nuclear transporters, followed by translocation through the nuclear envelope-embedded nuclear pore complexes (NPCs). It is becoming increasingly apparent, however, that distinct alternative pathways for nuclear transport exist and are relatively abundant. This review examines several of these novel pathways, including facilitation of Imp-dependent transport by microtubule motors, and Imp-independent pathways involving either other transport molecules such as the calcium-binding protein calmodulin or through direct binding to the components of the NPC. The existence of these pathways and the fact that many proteins appear to possess separate Imp-dependent and -independent nuclear import mechanisms ensure that the cell can function under conditions in which Imp-dependent transport is inhibited and/or modulate the efficiency of Imp dependent transport itself, according to the need. PMID- 19548982 TI - Protein kinase D-dependent trafficking of the large Herpes simplex virus type 1 capsids from the TGN to plasma membrane. AB - The biosynthetic pathway carries cargos from the endoplasmic reticulum (ER) to the trans Golgi network (TGN) via a typical passage through the Golgi. Interestingly, large particles such as procollagen, chylomicrons and some viruses all reach the TGN by atypical routes. Given this dichotomy, we anticipated that such cargos might rely on non-classical machineries downstream of the TGN. Using Herpes simplex virus type 1 (HSV-1) as a model and a synchronized infection protocol that focuses on TGN to plasma membrane transport, the present study revealed the surprising implication of the cellular serine-threonine protein kinase D in HSV-1 egress. These findings, confirmed by a variety of complementary means [pharmacological inhibitors, dominant negative mutant, RNA interference and electron microscopy (EM)], identify one of possibly several cellular factors that modulate the egress of viruses transiting at the TGN. Moreover, the involvement of this kinase, previously known to regulate the transport of small basolateral cargos, highlights the trafficking of both small and exceptionally large entities by a common machinery downstream of the TGN, in sharp contrast to earlier steps of transport. Conceptually, this indicates the TGN is not only a sorting station from which cargos can depart towards different destinations but also a meeting point where conventional and unconventional routes can meet along the biosynthetic pathway. Lastly, given the apical release of HSV-1 in neurons, it opens up the possibility that this kinase might regulate some apical sorting. PMID- 19548984 TI - Connexin 33 impairs gap junction functionality by accelerating connexin 43 gap junction plaque endocytosis. AB - Connexin 33 (Cx33) is a testis-specific gap junction protein. We previously reported that Cx33 exerts dominant-negative effect on gap junction intercellular communication by sequestering Cx43 within early endosomes in Sertoli cells. However, the molecular mechanisms that drive this process are unknown. The present study analyzed: (i) the trafficking of Cx33 and Cx43 in wild-type Sertoli cells transfected with Cx33-DsRed2 and Cx43-green fluorescent protein vectors; (ii) the formation of heteromeric Cx33/Cx43 hemi-channels and their incorporation into gap junction plaques. Fluorescence lifetime imaging microscopy-fluorescence resonance energy transfer and videomicroscopy studies demonstrated that Cx33 and Cx43 associated to form heteromeric oligomers that trafficked along microtubules to the plasma membrane. However, the plaques containing Cx33 were not functional. Immunoprecipitation experiments revealed that zonula occludens-1 (ZO-1), a scaffold protein proposed to secure Cx in gap junction plaques at the cell-cell boundary, associated with Cx33 in testis extracts. In cells expressing Cx33, Cx33 and ZO-1 specifically interacted with P(1) phosphorylated and P(0) unphosphorylated isoforms of Cx43, and the ZO-1 membranous signal level was reduced. It is suggested that alteration of Cx43/ZO-1 association by Cx33 could be one mechanism by which Cx33 exerts its dominant-negative effect on gap junction plaque. PMID- 19548985 TI - Binding site distribution of nuclear transport receptors and transport complexes in single nuclear pore complexes. AB - Transport through the nuclear pore complex (NPC) involves a large channel and an abundance of binding sites for nuclear transport receptors (NTRs). However, the mechanistically important distribution of NTR-binding sites along the channel is vividly debated. In this study, we visualized binding site distributions directly by two complementary optical super-resolution methods, single-molecule microscopy and 4Pi microscopy. First, we analyzed the distribution of RanGDP because this important nuclear transport substrate has two types of binding sites at the NPC, direct and indirect, NTR-mediated sites. We found that the direct binding sites had a maximum at approximately -30 nm with regard to the NPC center, whereas the indirect transport-relevant binding sites peaked at approximately -10 nm. The 20 nm-shift could be only resolved by 4Pi microscopy because of a two to threefold improved localization precision as compared with single-molecule microscopy. Then we analyzed the distribution of the NTR Kapbeta1 and a Kapbeta1-based transport complex and found them to have also binding maxima at approximately -10 nm. These observations support transport models in which NTR binding sites are distributed all along the transport channel and argue against models in which the cytoplasmic entrance of the channel is surrounded by a large cloud of binding sites. PMID- 19548986 TI - An acute severe ehrlichiosis in a dog experimentally infected with a new virulent strain of Ehrlichia canis. PMID- 19548988 TI - Molecular detection of Rickettsia typhi and Rickettsia felis in fleas from Algeria. PMID- 19548989 TI - First detection of Rickettsia aeschlimannii in Hyalomma aegyptium from Algeria. PMID- 19548990 TI - Rickettsia felis and Rickettsia massiliae in Ivory Coast, Africa. PMID- 19548991 TI - Role of the cytoplasmic pattern recognition receptor Nod2 in Coxiella burnetii infection. PMID- 19548992 TI - Rickettsia raoultii and Anaplasma phagocytophilum in Dermacentor reticulatus ticks collected from Bialowieza Primeval Forest European bison (Bison bonasus bonasus), Poland. PMID- 19548993 TI - Genotyping Rickettsia conorii detected in patients with Mediterranean spotted fever in Algeria using multispacer typing (MST). PMID- 19548994 TI - Coinfection with Coxiella burnetii in infectious endocarditis. PMID- 19548995 TI - Diagnosis of Coxiella burnetii pericarditis by using a systematic prescription kit in cases of pericardial effusion: an 8-year experience. PMID- 19548996 TI - Immunohistochemical detection of Coxiella burnetii in an aortic graft. PMID- 19548997 TI - Immunohistochemical detection of Coxiella burnetii in chronic Q fever hepatitis. PMID- 19548998 TI - Uveitis: an emerging clinical form of Bartonella infection. PMID- 19548999 TI - Clinical guidelines for interstitial cystitis and hypersensitive bladder syndrome. AB - A clinical guideline and algorism for interstitial cystitis and hypersensitive bladder syndrome has been developed by a group of East Asian urologists as a revised form of the Japanese guideline for interstitial cystitis. The guideline defines interstitial cystitis (IC) as a disease of the urinary bladder diagnosed by 3 requirements; 1) a characteristic complex of lower urinary tract symptoms, 2) bladder pathology such as Hunner's ulcer and bladder bleeding after overdistension, and 3) exclusions of confusable diseases. The characteristic symptom complex is termed as hypersensitive bladder syndrome (HBS), which is defined as bladder hypersensitivity, usually associated with urinary frequency, with or without bladder pain. For the definite diagnosis of IC, cytoscopy or hydrodistension is crutial; HBS is the diagnosis when IC is suspected but not confirmed by the 3 requirements. Numerous therapeutic options are available; however, most of them lack in high level of evidence, leaving a few as recommended therapies. Etiology of IC are multifactorial; the interaction among nervous, immune and endocrine factors forms a vicious cycle, provocating and maintaining inflammatory reactions in the bladder. The inclusion and efficacy criteria for clinical trials should be standardized to enhance the clinical research for this disabling disease, which has proved to be more prevalent than previously believed. PMID- 19549000 TI - Virological evaluation of domestic water purification devices commonly used in India emphasizes inadequate quality and need for virological standards. AB - OBJECTIVES: To evaluate the performance of domestic water purification units with respect to contaminating enteric viruses. METHODS: Eight domestic water purification systems widely used in India were evaluated using hepatitis E virus (HEV) as a model virus. For HEV concentration and detection, membrane filtration and real-time PCR were used respectively. Viral log reduction value (LRV) was calculated for each unit. RESULTS: Viral log reduction value was 0.21 for unit 6 (polyester + carbon), 1.45 for unit 4 (filter + UV), 1.52 for unit 3 (filter + chlorine), 1.70 for a carbon + exhaust indication contact disinfection unit, 2.20 for an iodine resin unit, 2.51 for a dual filter unit and 6.53 for a hollow fibre membrane unit and a gravity-fed filter unit. Thus, only the technologies employed by the latter two were efficient in complete removal of HEV. CONCLUSIONS: The majority of the water purifiers under use are inadequate. Virological standards in evaluating such devices need to be established urgently, in order to help manufacturers to improve the performance of such products and most importantly, to help consumers. PMID- 19549001 TI - Adherence to artesunate-amodiaquine combination therapy for uncomplicated malaria in children in Zanzibar, Tanzania. AB - OBJECTIVES: To estimate caretaker adherence to co-blistered, but not co formulated, artesunate-amodiaquine (AsAq) for uncomplicated malaria and identify factors associated with caretaker adherence. METHODS: Cross sectional household survey of caretakers of 210 children under 5 years of age who had been prescribed and dispensed AsAq at 21 public health facilities (HFs). The caretakers were interviewed in their homes on the 4th day of receiving the 3 day treatment. Adherence of caretakers was assessed by self report and pill count. RESULTS: Caretaker adherence to AsAq was 77% (95% CI: 67%-87%). Non-adherence resulted in under-dosing (3/4) of the time and was most often in the form of wrong daily doses due to misunderstanding or forgetting the correct dose regimens. Predictors of adherence were education exceeding 7 years (OR = 5.08, P = 0.008) and receiving the exact number of pills to complete the treatment regimen (OR = 4.09, P = 0.006). All caretakers of children who were administered the first dose at the HF had adhered to the treatment. CONCLUSION: We found moderate levels of caretaker adherence to AsAq. Further improvement could be achieved by producing dose-specific packaging for infants, providing clear instructions and giving the first dose under observation at the HF. PMID- 19549002 TI - Melatonin improves cardiovascular function and ameliorates renal, cardiac and cerebral damage in rats with renovascular hypertension. AB - The effect of melatonin was investigated in an angiotensin II-dependent renovascular hypertension model in Wistar albino rats by placing a renal artery clip (two-kidney, one-clip; 2K1C), while sham rats did not have clip placement. Starting either on the operation day or 3 wk after the operation, the rats received melatonin (10 mg/kg/day) or vehicle for the following 6 wk. At the end of the nineth week, after blood pressure (BP) and echocardiographic recordings were obtained, plasma samples were obtained to assay lactate dehydrogenase (LDH), creatine kinase (CK), antioxidant capacity (AOC), asymmetric dimethylarginine (ADMA), and nitric oxide (NOx) levels. In the kidney, heart and brain tissues, malondialdehyde (MDA) and glutathione (GSH) levels, superoxide dismutase (SOD), catalase (CAT), myeloperoxidase (MPO) and Na(+)-K(+) ATPase activities were determined. 2K1C caused an increase in BP and left ventricular (LV) dysfunction. In hypertensive animals LDH, CK, ADMA levels were increased in plasma with a concomitant reduction in AOC and NOx. Moreover, hypertension caused a significant decrease in tissue SOD, CAT, and Na(+), K(+)-ATPase activities and glutathione content, while MDA levels and MPO activity were increased in all studied tissues. On the other hand, both melatonin regimens significantly reduced BP, alleviated oxidative injury and improved LV function. In conclusion, melatonin protected against renovascular hypertension-induced tissue damage and improved cardiac function presumably due to both its direct antioxidant and receptor-dependent actions, suggesting that melatonin may be of therapeutic use in preventing oxidative stress due to hypertension. PMID- 19549003 TI - Iliac cortical thickness in the neonate - the gradient effect. AB - Recent studies of the neonatal ilium are beginning to reveal that a recognizable structural patterning of trabecular bone is present in the absence of any direct stance-related weight transfer. However, little is known about the organization of compact bone in the ilium and the way in which it is laid down during the earliest stages of development. This study investigates cortical bone thickness across both gluteal and pelvic iliac shells in the human neonatal ilium. Measurements of specific regions of interest on the iliac cortices were recorded using reconstructed micro-computed tomography scans from 30 neonatal ilia. Analysis of gluteal and pelvic cortical thicknesses revealed a distinctive patterning consistent with the expected bone distribution achieved through early bone modelling and remodelling. The analysis of this pattern is important for understanding the relationship between trabecular bone patterning and cortical bone structure in the earliest stages of pelvic development prior to locomotive influences and its response to the specific functional forces acting during this period. PMID- 19549004 TI - Linked morphological changes during palate evolution in early tetrapods. AB - We examined the shapes and sizes of dermal bones of the palate of selected Palaeozoic tetrapods in order to identify the ancestral states of palatal bone morphologies in the earliest tetrapods, to learn how the composition of the palate varies within and among early tetrapod radiations, and to recognize evolutionary correlations among the size and shapes of skeletal elements in this important group of animals. We find that whereas the palatal bones themselves and their arrangements are usually conserved, considerable correlated evolutionary change occurs in the shapes and sizes of the bones. Some of the changes in the bones are allometrically linked to overall palate size, which varies more than 100-fold among the taxa in our sample. Often, these allometries were only hinted at in traditional independent contrasts-based regressions of log transformed data, particularly because many allometries are subtle, their slopes may vary among subclades, and the scatter around some trendlines is high. Rather, the allometries showed up in analyses of size-standardized palatal bone dimensions investigated using independent contrasts, bivariate phylomorphospace plots, and mirrored character reconstructions on the phylogenetic tree. We find negative allometry for parasphenoid lengths and widths essentially across the entire tree of Palaeozoic tetrapods, but with different trajectories characterizing the two largest clades, the temnospondyls and the lepospondyls. The lengths of several other elements may show positive allometries, either across the entire tree or in just a subclade. One possible positive allometry exists for the ectopterygoid, which appears to shorten allometrically in temnospondyls that evolve small body and palate size, and, as in Doleserpeton can be lost altogether. Both shortening and loss could be by the same developmental change, paedomorphosis, a form of heterochrony. Paedomorphosis might also account for evolution of relatively large parasphenoids in both lepospondyls and diminutive temnospondyls, but does not seem to explain evolution of ectopterygoid loss in lepospondyls. A regularity observed across nearly all taxa in our study set is an inverse correlation between the lengths of the vomer and pterygoid, bones that lie adjacent to one another along the long palatal axis. Further work is needed to learn whether such correlated evolution might be due to adaptation and/or to developmental bias, and particularly to learn how correlations and allometries themselves evolve. PMID- 19549006 TI - Relationship between microglial activation and dopaminergic neuronal loss in the substantia nigra: a time course study in a 6-hydroxydopamine model of Parkinson's disease. AB - Cellular interactions between activated microglia and degenerating neurons in in vivo models of Parkinson's disease are not well defined. This time course study assesses the dynamics of morphological and immunophenotypic properties of activated microglia in a 6-hydroxydopamine (6-OHDA) model of Parkinson's disease. Neurodegeneration in the substantia nigra pars compacta (SNc) was induced by unilateral injection of 6-OHDA into the medial forebrain bundle. Activated microglia, identified using monoclonal antibodies: clone of antibody that detects major histocompatibility complex (MHC) class II antigens (OX6) for MHC class II, clone of antibody that detects cell surface antigen-cluster of differentiation 11b - anti-complement receptor 3, a marker for complement receptor 3 and CD 68 for phagocytic activity. Activation of microglia in the lesioned SNc was rapid with cells possessing amoeboid or ramified morphology appeared on day 1, whilst antibody clone that detects macrophage-myeloid associated antigen immunoreactivity was observed at day 3 post-lesion when there was no apparent loss of tyrosine hydroxylase (TH)+ve dopaminergic (DA) SNc neurons. Thereafter, OX6 and antibody clone that detects macrophage-myeloid associated antigen activated microglia selectively adhered to degenerating axons, dendrites and apoptotic (caspase 3+ve) DA neurons in the SNc were observed at day 7. This was followed by progressive loss of TH+ve SNc neurons, with the peak of TH+ve cell loss (51%) being observed at day 9. This study suggests that activation of microglia precedes DA neuronal cell loss and neurons undergoing degeneration may be phagocytosed prematurely by phagocytic microglia. PMID- 19549007 TI - L-aspartate as an amino acid neurotransmitter: mechanisms of the depolarization induced release from cerebrocortical synaptosomes. AB - The role of L-aspartate as a classical neurotransmitter of the CNS has been a matter of great debate. In this study, we have characterized the main mechanisms of its depolarization-induced release from rat purified cerebrocortical synaptosomes in superfusion and compared them with those of the well known excitatory neurotransmitter L-glutamate. High KCl and 4-aminopyridine were used as depolarizing agents. At 15 mM KCl, the overflows of both transmitters were almost completely dependent on external Ca2+. At 35 and 50 mM KCl, the overflows of L-aspartate, but not those of L-glutamate, became sensitive to DL-threo-b benzyloxy aspartic acid (DL-TBOA), an excitatory amino acid transporter inhibitor. In the presence of DL-TBOA, the 50 mM KCl-evoked release of L aspartate was still largely external Ca2+-dependent. The DL-TBOA insensitive,external Ca2+-independent component of the 50 mM KCl-evoked overflows of L-aspartate and L-glutamate was significantly decreased by the mitochondrial Na+/Ca2+ exchanger blocker CGP 37157. The Ca2+-dependent, KCl-evoked overflows of L-aspartate and L-glutamate were diminished by botulinum neurotoxin C, although to a significantly different extent. The 4-aminopyridine-induced L-aspartate and L-glutamate release was completely external Ca2+-dependent and never affected by DL-TBOA. Superimposable results have been obtained by pre-labeling synaptosomes with [3H]D aspartate and [3H]L-glutamate. Therefore, our data showing that L aspartate is released from nerve terminals by calcium dependent,exocytotic mechanisms support the neurotransmitter role of this amino acid. PMID- 19549008 TI - F-spondin plays a critical role in murine neuroblastoma survival by maintaining IL-6 expression. AB - F-spondin is associated with the regulation of axonal growth and the development of the nervous system. Its mechanism of action, however, is not clearly understood. In this study, we found that murine neuroblastoma Neuro-2a cells expressed a significant level of IL-6, but only trace amounts of IL-12, tumor necrosis factor alpha and nitric oxide. Knock-down of F-spondin mRNA in murine neuroblastoma NB41A3 and Neuro-2a cells using small interfering RNAs led to decreased IL-6 levels along with lower resistance to serum starvation and cytotoxic amyloid beta(1-42) (Abeta(1-42)) peptide. Restoring decline of F spondin or IL-6 induced by F-spondin knock-down through adding exogenous F spondin, IL-6 or over-expressing F-spondin reversed the cell death induced by Abeta(1-42) peptide or serum starvation. The decrease of IL-6 level was positively correlated with decrease of NF-kappaB and inhibition of p38 mitogen activated protein kinase (MAPK). Over-expressing MEKK, a kinase activator of the p38 MAPK pathway, increased IL-6 production, restored the decrease of p38 induced by F-spondin knock-down, and rescued the cells from death caused by Abeta(1-42) peptide. Taken together, these results suggest that F-spondin may play a critical role in murine neuroblastoma survival under adverse conditions by maintaining IL 6 level via a MEKK/p38 MAPK/NF-kappaB-dependent pathway. PMID- 19549009 TI - Increased vulnerability of hippocampal pyramidal neurons to the toxicity of kainic acid in OASIS-deficient mice. AB - The endoplasmic reticulum (ER) stress response is a defense system for dealing with the accumulation of unfolded proteins in the ER lumen. Old astrocyte specifically induced substance (OASIS) is known to be expressed in astrocytes and involved in the ER stress response; however the function of OASIS in the injured brain has remained unclear. In this study, we examined the roles of OASIS in neuronal degeneration in the hippocampi of mice intraperitoneally injected with kainic acid (KA). OASIS mRNA was strongly induced in response to KA injection, with a similar time course to the induction of ER molecular chaperone immunoglobulin heavy chain binding protein mRNA. In situ hybridization showed that KA injection causes induction of immunoglobulin heavy chain binding protein mRNA in glial fibrillary acidic protein-positive astrocytes as well as in pyramidal neurons, although up-regulation of OASIS mRNA was only detected in glial fibrillary acidic protein-positive astrocytes. Primary cultured astrocytes, but not the neurons of OASIS-/- mice, revealed reduced vulnerability to ER stress. Furthermore, pyramidal neurons in the hippocampi of OASIS-/- mice were more susceptible to the toxicity induced by KA than those of wild-type mice. Taken together, these data suggest that OASIS expressed in astrocytes plays important roles in protection against the neuronal damage induced by KA. PMID- 19549010 TI - The association between emergency department crowding and adverse cardiovascular outcomes in patients with chest pain. AB - OBJECTIVES: While emergency department (ED) crowding is a worldwide problem, few studies have demonstrated associations between crowding and outcomes. The authors examined whether ED crowding was associated with adverse cardiovascular outcomes in patients with chest pain syndromes (chest pain or related complaints of possible cardiac origin). METHODS: A retrospective analysis was performed for patients >or=30 years of age with chest pain syndrome admitted to a tertiary care academic hospital from 1999 through 2006. The authors compared rates of inpatient adverse outcomes from ED triage to hospital discharge, defined as delayed acute myocardial infarction (AMI), heart failure, hypotension, dysrhythmias, and cardiac arrest, which occurred after ED arrival using five separate crowding measures. RESULTS: Among 4,574 patients, 251 (4%) patients developed adverse outcomes after ED arrival; 803 (18%) had documented acute coronary syndrome (ACS), and of those, 273 (34%) had AMI. Compared to less crowded times, ACS patients experienced more adverse outcomes at the highest waiting room census (odds ratio [OR] = 3.7, 95% confidence interval [CI] = 1.3 to 11.0) and patient hours (OR = 5.2, 95% CI = 2.0 to 13.6) and trended toward more adverse outcomes during time of high ED occupancy (OR = 3.1, 95% CI = 1.0 to 9.3). Adverse outcomes were not significantly more frequent during times with the highest number of admitted patients (OR = 1.6, 95% CI = 0.6 to 4.1) or the highest trailing mean length of stay (LOS) for admitted patients transferred to inpatient beds within 6 hours (OR = 1.5, 95% CI = 0.5 to 4.0). Patients with non-ACS chest pain experienced more adverse outcomes during the highest waiting room census (OR = 3.5, 95% CI = 1.4 to 8.4) and patient-hours (OR = 4.3, 95% CI = 2.6 to 7.3), but not occupancy (OR = 1.8, 95% CI = 0.9 to 3.3), number of admitted patients (OR = 0.6, 95% CI 0.4 to 1.1), or trailing LOS for admitted patients (OR = 1.2, 95% CI = 0.6 to 2.0). CONCLUSIONS: There was an association between some measures of ED crowding and a higher risk of adverse cardiovascular outcomes in patients with both ACS-related and non-ACS-related chest pain syndrome. PMID- 19549012 TI - Diagnosis of acute mitral valve insufficiency using emergency bedside echocardiography. PMID- 19549013 TI - Emergency department diagnosis of aortic dissection by bedside transabdominal ultrasound. PMID- 19549014 TI - Financial impact of emergency department ultrasound. AB - OBJECTIVES: There is limited information on the financial implications of an emergency department ultrasound (ED US) program. The authors sought to perform a fiscal analysis of an integrated ED US program. METHODS: A retrospective review of billing data was performed for fiscal year (FY) 2007 for an urban academic ED with an ED US program. The ED had an annual census of 80,000 visits and 1,101 ED trauma activations. The ED is a core teaching site for a 4-year emergency medicine (EM) residency, has 35 faculty members, and has 24-hour availability of all radiology services including formal US. ED US is utilized as part of evaluation of all trauma activations and for ED procedures. As actual billing charges and reimbursement rates are institution-specific and proprietary information, relative value units (RVUs) and reimbursement based on the Centers for Medicare & Medicaid Services (CMS) 2007 fee schedule (adjusted for fixed diagnosis-related group [DRG] payments and bad debt) was used to determine revenue generated from ED US. To estimate potential volume, assumptions were made on improvement in documentation rate for diagnostic scans (current documentation rates based on billed volume versus diagnostic studies in diagnostic image database), with no improvements assumed for procedural ED US. Expenses consist of three components-capital costs, training costs, and ongoing operational costs-and were determined by institutional experience. Training costs were considered sunken expenses by this institution and were thus not included in the original return on investment (ROI) calculation, although for this article a second ROI calculation was done with training cost estimates included. For the purposes of analysis, certain key assumptions were made. We utilized a collection rate of 45% and hospitalization rates (used to adjust for fixed DRG payments) of 33% for all diagnostic scans, 100% for vascular access, and 10% for needle placement. An optimal documentation rate of 95% was used to estimate potential revenue. RESULTS: In FY 2007, 486 limited echo exams of abdomen (current procedural terminology [CPT] 76705) and 480 limited echo cardiac exams were performed (CPT 93308) while there were 78 exams for US-guided vascular access (CPT 76937) and 36 US-guided needle placements when performing paracentesis, thoracentesis, or location of abscess for drainage (CPT 76492). Applying the 2007 CMS fee schedule and above assumptions, the revenue generated was 578 RVUs and $35,541 ($12,934 in professional physician fees and $22,607 in facility fees). Assuming optimal documentation rates for diagnostic ED US scans, ED US could have generated 1,487 RVUs and $94,593 ($33,953 in professional physician fees and $60,640 in facility fees). Program expenses include an initial capital expense (estimated at $120,000 for two US machines) and ongoing operational costs ($68,640 per year to cover image quality assurance review, continuing education, and program maintenance). Based on current revenue, there would be an annual operating loss, and thus an ROI cannot be calculated. However, if potential revenue is achieved, the annual operating income will be $22,846 per year with an ROI of 4.9 years to break even with initial investment. CONCLUSIONS: Determining an ROI is a required procedure for any business plan for establishing an ED US program. Our analysis demonstrates that an ED US program that captures charges for trauma and procedural US and achieves the potential billing volume breaks even in less than 5 years, at which point it would generate a positive margin. PMID- 19549015 TI - The impact of injury coding schemes on predicting hospital mortality after pediatric injury. AB - OBJECTIVES: Accurate adjustment for injury severity is needed to evaluate the effectiveness of trauma management. While the choice of injury coding scheme used for modeling affects performance, the impact of combining coding schemes on performance has not been evaluated. The purpose of this study was to use Bayesian logistic regression to develop models predicting hospital mortality in injured children and to compare the performance of models developed using different injury coding schemes. METHODS: Records of children (age < 15 years) admitted after injury were obtained from the National Trauma Data Bank (NTDB) and the National Pediatric Trauma Registry (NPTR) and used to train Bayesian logistic regression models predicting mortality using three injury coding schemes (International Classification of Disease-9th revision [ICD-9] injury codes, the Abbreviated Injury Scale [AIS] severity scores, and the Barell matrix) and their combinations. Model performance was evaluated using independent data from the NTDB and the Kids' Inpatient Database 2003 (KID). RESULTS: Discrimination was optimal when modeling both ICD-9 and AIS severity codes (area under the receiver operating curve [AUC] = 0.921 [NTDB] and 0.967 [KID], Hosmer-Lemeshow [HL] h statistic = 115 [NTDB] and 147 [KID]), while calibration was optimal when modeling coding based on the Barell matrix (AUC = 0.882 [NTDB] and 0.936 [KID], HL h-statistic = 19 [NTDB] and 69 [KID]). When compared to models based on ICD-9 codes alone, models that also included AIS severity scores and coding from the Barell matrix showed improved discrimination and calibration. CONCLUSIONS: Mortality models that incorporate additional injury coding schemes perform better than those based on ICD-9 codes alone in the setting of pediatric trauma. Combining injury coding schemes may be an effective approach for improving the predictive performance of empirically derived estimates of injury mortality. PMID- 19549016 TI - Prospective validation of the pediatric appendicitis score in a Canadian pediatric emergency department. AB - OBJECTIVES: Clinical scoring systems attempt to improve the diagnostic accuracy of pediatric appendicitis. The Pediatric Appendicitis Score (PAS) was the first score created specifically for children and showed excellent performance in the derivation study when administered by pediatric surgeons. The objective was to validate the score in a nonreferred population by emergency physicians (EPs). METHODS: A convenience sample of children, 4-18 years old presenting to a pediatric emergency department (ED) with abdominal pain of less than 3 days' duration and in whom the treating physician suspected appendicitis, was prospectively evaluated. Children who were nonverbal, had a previous appendectomy, or had chronic abdominal pathology were excluded. Score components (right lower quadrant and hop tenderness, anorexia, pyrexia, emesis, pain migration, leukocytosis, and neutrophilia) were collected on standardized forms by EPs who were blinded to the scoring system. Interobserver assessments were completed when possible. Appendicitis was defined as appendectomy with positive histology. Outcomes were ascertained by review of the pathology reports from the surgery specimens for children undergoing surgery and by telephone follow-up for children who were discharged home. Sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) were calculated. The overall performance of the score was assessed by a receiver operator characteristic (ROC) curve. RESULTS: Of the enrolled children who met inclusion criteria (n = 246), 83 (34%) had pathology-proven appendicitis. Using the single cut-point suggested in the derivation study (PAS 5) resulted in an unacceptably high number of false positives (37.6%). The score's performance improved when two cut-points were used. When children with a PAS of or=8 determined the need for appendectomy, the score's specificity was 95.1% with a PPV of 85.2%. Using this strategy, the negative appendectomy rate would have been 8.8%, the missed appendicitis rate would have been 2.4%, and 41% of imaging investigations would have been avoided. CONCLUSIONS: The PAS is a useful tool in the evaluation of children with possible appendicitis. Scores of or=8 help predict appendicitis. Patients with a PAS of 5-7 may need further radiologic evaluation. PMID- 19549017 TI - Cocaine and ethanol: combined effects on coronary artery blood flow and myocardial function in dogs. AB - OBJECTIVES: In combination, cocaine and ethanol are more cardiotoxic than is either substance alone. These substances together constitute a drug abuse combination that commonly results in fatality. Previously the authors have demonstrated that cardiotoxicity of cocaine and ethanol is in part due to synergistic myocardial-depressant effects. However, it remains unclear whether this myocardial depression is associated with concomitant adverse effects on coronary blood flow in relation to these substances. The aim of this study was to investigate combined effects of cocaine and ethanol on myocardial blood flow, in relation to indices of myocardial function. METHODS: Anesthetized dogs were instrumented for hemodynamic monitoring with Doppler flow probes placed on the circumflex and left anterior descending (LAD) coronary arteries. Dogs were randomized to three groups (each n = 6): ethanol (E, 1.5 g/kg followed by placebo), cocaine (C, placebo followed by cocaine, 7.5 mg/kg IV), or cocaine plus ethanol (C + E). All measurements were made at control, after placebo or ethanol, and then at fixed time intervals after cocaine or placebo bolus over 3 hours. RESULTS: In both the C + E and the C groups, circumflex blood flow (CBF) decreased by 71% (95% confidence interval [CI] = 56% to 85%) and 57% (95% CI = 43% to 72%, both p < 0.04 vs. baseline) immediately after cocaine bolus. This was associated with transient depression of cardiac output, myocardial contractile function, and rate-pressure product (RPP), all indices of myocardial oxygen demand. A subsequent rebound increase of coronary sinus blood flow (CSBF) of 56% (95% CI = 26% to 137%, p < 0.03) compared to baseline occurred only in the C group and was associated with increases of myocardial contractile function and RPP. In the C + E group, 2 hours after drug administration, there was a decrease in CSBF of 49% (95% CI = 32% to 67%; p < 0.01) compared to baseline, which was associated with concomitant numerical decreases of the indices of myocardial oxygen demand and accumulation of cocaethylene. CONCLUSIONS: Acute decreases in myocardial flow secondary to cocaine, and cocaine and ethanol in combination, were similar and temporally associated with cocaine's direct myocardial depressant effects. Rebound increases in myocardial function and blood flow due to cocaine were attenuated by ethanol. Delayed myocardial depression and decreases in myocardial blood flow were observed only with coadministration of cocaine and ethanol. PMID- 19549018 TI - The association between emergency department crowding and analgesia administration in acute abdominal pain patients. AB - OBJECTIVES: The authors assessed the effect of emergency department (ED) crowding on the nontreatment and delay in treatment for analgesia in patients who had acute abdominal pain. METHODS: This was a secondary analysis of prospectively enrolled nonpregnant adult patients presenting to an urban teaching ED with abdominal pain during a 9-month period. Each patient had four validated crowding measures assigned at triage. Main outcomes were the administration of and delays in time to analgesia. A delay was defined as waiting more than 1 hour for analgesia. Relative risk (RR) regression was used to test the effects of crowding on outcomes. RESULTS: A total of 976 abdominal pain patients (mean [+/-standard deviation] age = 41 [+/-16.6] years; 65% female, 62% black) were enrolled, of whom 649 (67%) received any analgesia. Of those treated, 457 (70%) experienced a delay in analgesia from triage, and 320 (49%) experienced a delay in analgesia after room placement. After adjusting for possible confounders of the ED administration of analgesia (age, sex, race, triage class, severe pain, final diagnosis of either abdominal pain not otherwise specified or gastroenteritis), increasing delays in time to analgesia from triage were independently associated with all four crowding measures, comparing the lowest to the highest quartile of crowding (total patient-care hours RR = 1.54, 95% confidence interval [CI] = 1.32 to 1.80; occupancy rate RR = 1.64, 95% CI = 1.42 to 1.91; inpatient number RR = 1.57, 95% CI = 1.36 to 1.81; and waiting room number RR = 1.53, 95% CI = 1.31 to 1.77). Crowding measures were not associated with the failure to treat with analgesia. CONCLUSIONS: Emergency department crowding is associated with delays in analgesic treatment from the time of triage in patients presenting with acute abdominal pain. PMID- 19549019 TI - Identifying star performers: the relationship between ambitious targets and nursing home quality improvement. AB - Setting Targets--Achieving Results (STAR) is a Web-based tool that helps nursing home leadership select annual performance goals, or targets, for a subset of publicly reported quality measures. Previous results demonstrate that nursing homes whose staff implement STAR targets demonstrate greater improvement on the related outcomes. In this analysis, the authors hypothesized that nursing homes whose staff select the most ambitious targets (reflecting large improvement over their current performance) may be more successful in their related quality improvement efforts than homes with less-ambitious targets (reflecting lesser improvement). The authors analyzed data from 7,091 Medicare- or Medicaid certified nursing homes that set STAR targets in 2005 or 2006 for two quality measures: the proportion of residents who were physically restrained daily and the proportion of high-risk residents with pressure ulcers. Targets were classified as ambitious or less ambitious based on the 75th and 50th rank-ordered percentiles, respectively. Improvement was calculated using four-quarter averages for baseline (the year ending when the target was set) and remeasurement (the subsequent year). The results indicate that nursing homes with ambitious targets demonstrate greater improvement than their peers selecting less-ambitious targets. With limited federal and local resources to assist providers with quality improvement, target values may be a used as a "flag" to help agencies allocate scarce resources to nursing homes committed to quality improvement efforts and with the organizational capacity to improve. PMID- 19549020 TI - Effect of fish oil supplementation on quality of life in a general population of older Dutch subjects: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVES: To investigate the effect of eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) supplementation on quality of life (QOL). DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: Independently living individuals from the general older Dutch population. PARTICIPANTS: Three hundred two individuals aged 65 and older without depression or dementia. INTERVENTION: 1,800 mg/d EPA-DHA (n=96), 400 mg/d EPA-DHA (n=100), or placebo capsules (n=106) for 26 weeks. MEASUREMENTS: QOL was assessed using the short version of the World Health Organization QOL questionnaire (WHOQOL-BREF). The WHOQOL-BREF covers four domains: physical health, psychological health, social relationships, and satisfaction with environment. The total score range is 26 to 130, with higher scores indicating a more favorable condition. RESULTS: Mean age of the participants was 70, and 55% were male. Plasma concentrations of EPA-DHA increased 238% in the high-dose and 51% in the low-dose EPA-DHA group, reflecting excellent adherence. Median baseline total WHOQOL scores ranged from 107 to 110 in the three groups and were not significantly different from each other. After 26 weeks, the mean difference from placebo was -1.42 (95% confidence interval (CI)=-3.40-0.57) for the high-dose and 0.02 (95% CI=-1.95-1.99) for the low-dose fish oil group. Treatment with 1,800 mg or 400 mg EPA-DHA did not affect total QOL or any of the separate domains after 13 or 26 weeks of intervention. CONCLUSION: Supplementation with high or low doses of fish oil for 26 weeks did not influence the QOL of healthy older individuals. PMID- 19549021 TI - Prospective study of serum 25-hydroxyvitamin D level, cardiovascular disease mortality, and all-cause mortality in older U.S. adults. AB - OBJECTIVES: To evaluate the association between serum 25-hydroxyvitamin D (25(OH)D) levels and mortality in a representative U.S. sample of older adults. DESIGN: Prospective cohort from the Third National Health and Nutrition Examination Survey (NHANES III) and linked mortality files. SETTING: Noninstitutionalized U.S. civilian population. PARTICIPANTS: Three thousand four hundred eight NHANES III participants aged 65 and older enrolled from 1988 to 1994 and followed for mortality through 2000. MEASUREMENTS: Primary exposure was serum 25(OH)D level at enrollment. Primary and secondary outcomes were all-cause and cardiovascular disease (CVD) mortality, respectively. RESULTS: During the median 7.3 years of follow-up, there were 1,493 (44%) deaths, including 767 CVD related deaths. Median 25(OH)D level was 66 nmol/L. Adjusting for demographics, season, and cardiovascular risk factors, baseline 25(OH)D levels were inversely associated with all-cause mortality risk (adjusted hazard ratio (HR)=0.95, 95% confidence interval (CI)=0.92-0.98, per 10 nmol/L 25[OH]D). Compared with subjects with 25(OH)D levels of 100 nmol/L or higher, the adjusted HR for subjects with levels less than 25.0 nmol/L was 1.83 (95% CI=1.14-2.94) and for levels of 25.0 to 49.9 nmol/L was 1.47 (95% CI=1.09-1.97). The association appeared stronger for CVD mortality (adjusted HR=2.36, 95% CI=1.17-4.75, for subjects with 25[OH]D levels<25.0 nmol/L vs those > or =100.0 nmol/L) than for non-CVD mortality (adjusted HR=1.42, 95% CI=0.73-2.79, for subjects with 25[OH]D levels<25.0 nmol/L vs those > or =100.0 nmol/L). CONCLUSION: In noninstitutionalized older adults, a group at high risk for all-cause mortality, serum 25(OH)D levels had an independent, inverse association with CVD and all cause mortality. Randomized controlled trials of vitamin D supplementation in older adults are warranted to determine whether this association is causal and reversible. PMID- 19549022 TI - Computerized decision support to reduce potentially inappropriate prescribing to older emergency department patients: a randomized, controlled trial. AB - OBJECTIVES: To evaluate the effectiveness of computer-assisted decision support in reducing potentially inappropriate prescribing to older adults. DESIGN: Randomized, controlled trial. SETTING: An academic emergency department (ED) in Indianapolis, Indiana, where computerized physician order entry was used to write all medication prescriptions. PARTICIPANTS: Sixty-three emergency physicians were randomized to the intervention (32 physicians) or control (31 physicians) group. INTERVENTION: Decision support that advised against use of nine potentially inappropriate medications and recommended safer substitute therapies. MEASUREMENTS: The primary outcome was the proportion of ED visits by seniors that resulted in one or more prescriptions for an inappropriate medication. The main secondary outcomes were the proportions of medications prescribed that were inappropriate and intervention physicians' reasons for rejecting the decision support. RESULTS: The average age of the patients was 74, two-thirds were female, and just over half were African American. Decision support was provided 114 times to intervention physicians, who accepted 49 (43%) of the recommendations. Intervention physicians prescribed one or more inappropriate medications during 2.6% of ED visits by seniors, compared with 3.9% of visits managed by control physicians (P=.02; odds ratio=0.55, 95% confidence interval=0.34-0.89). The proportion of all prescribed medications that were inappropriate significantly decreased from 5.4% to 3.4%. The most common reason for rejecting decision support was that the patient had no prior problems with the medication. CONCLUSION: Computerized physician order entry with decision support significantly reduced prescribing of potentially inappropriate medications for seniors. This approach might be used in other efforts to improve ED care. TRIAL REGISTRATION: Clinical trials.gov Identifier: NCT00297869. PMID- 19549023 TI - Thirteen dimensions of health in elderly Sri Lankans: results from a National Sri Lanka Aging Survey. AB - OBJECTIVES: To explore age and sex differences in distribution of 13 health dimensions with a focus on self-rated health (SRH) and the association between SRH and other health dimensions in elderly Sri Lankans. DESIGN: Sri Lanka Aging Survey, a nationally representative cross-sectional survey. SETTING: Community based. PARTICIPANTS: Inhabitants of 13 districts in Sri Lanka aged 60 and older (N=2,413). MEASUREMENTS: Self-reported SRH, hearing, activities of daily living (ADLs), instrumental activities of daily living (IADLs), mobility, physical disability, chronic diseases, stress and worry, mood, cognition, social participation, social support, and financial health. RESULTS: The prevalence of being "healthy" in most health dimensions, including SRH, declined with age. Men were more likely to report better SRH; independence in ADLs, IADLs, and mobility; absence of physical disability and chronic diseases; and good mental health. Absence of chronic diseases, independence in ADLs and IADLs, freedom from stress and worry, and absence of depression were associated with positive (excellent/very good/good) SRH. The male SRH advantage was not significant in adjusted analyses, and sex did not modify the association between SRH and other health dimensions. CONCLUSION: Several of the dimensions associated with positive SRH are to a certain extent modifiable and therefore provide a potential for improvement in SRH of elderly Sri Lankans. Differences between this study and studies from elsewhere in the associations between different health dimensions and SRH, and in how sex modifies these associations, suggest that some of the associations may depend on cultural context. PMID- 19549024 TI - Budesonide and formoterol inhibit inflammatory mediator production by bronchial epithelial cells infected with rhinovirus. AB - BACKGROUND: Rhinoviruses (RVs) are responsible for the majority of acute asthma and chronic obstructive pulmonary disease (COPD) exacerbations. RVs infect the lower airways and induce the production of pro-inflammatory and remodelling associated mediators. Budesonide (BUD) and formoterol (FORM) synergize in controlling asthma and COPD exacerbations; however, their effects on virus induced inflammation and remodelling are less known. OBJECTIVE: We investigated whether BUD and FORM synergize in suppressing RV-induced inflammation and remodelling in the airways. METHODS: In vitro models of RV infection of BEAS-2B and primary normal human bronchial epithelial (NHBE) cells were used. We assessed the effects of individual and combined drugs administered post-infection, at a clinically relevant concentration range (10(-6)-10(-10) m), on the production of CCL5, CXCL10, CXCL8, IL-6 and the remodelling-associated VEGF and bFGF, using ELISA and RT-PCR. RESULTS: BUD effectively suppressed RV-mediated induction of all mediators studied, in a concentration-dependent manner. FORM alone suppressed the production of CXCL8 and bFGF. The combination of BUD and FORM had concentration-dependent, additive or synergistic effects in the suppression of RV induced CCL5, CXCL8 and CXCL10 in both cell types as well as VEGF in NHBE only. Combination treatment also resulted in an enhanced suppression of RV-induced IL 6, and CCL5 at the mRNA level as compared with BUD or FORM alone. CONCLUSION: BUD and FORM suppress RV-induced chemokines and growth factors in bronchial epithelial cells in a concentration-dependent, synergistic or additive manner. These data further support the combined use of BUD and FORM in asthma and COPD and intensification of this therapy during exacerbations. PMID- 19549025 TI - Changes in asthma-like responses after extended removal from exposure to trimellitic anhydride in the Brown Norway rat model. AB - BACKGROUND: Organic acid anhydride-induced occupational asthma is considered to be IgE-mediated. Airway and skin exposure are the two main routes of sensitization in the work place. Recently we developed an allergic asthmatic Brown Norway rat model sensitized by dermal exposure to trimellitic anhydride (TMA) using an occlusion patch application. OBJECTIVES: The objectives of this study were (1) to develop a model of non-occluded dermal exposure leading to allergic sensitization and (2) to examine the effect of extended removal from exposure on persistence of both specific IgE and TMA aerosol-induced airway responses in this model. METHODS: TMA powder (4 or 40 mg) was applied, unoccluded, to the skin of rats for 4 h, once/week for 4 weeks. Rats were given a 10-min aerosol challenge to 40 mg/m(3) TMA 2 weeks after the last dermal exposure (day 35). Another group was challenged on day 35 and again 18-24 months later. Respiratory enhanced pause (Penh), pulmonary histopathology and inflammation and specific IgE titres were measured. RESULTS: Rats produced dose-dependent specific IgE titres after exposure and developed early-phase (EAR) and late-phase airway responses (LAR) after airway challenge to TMA aerosol as well as airway eosinophilic inflammation. Specific airway responses were still manifested after a second TMA airway challenge given 18-24 months following the initial airway challenge. While persistent, airway inflammation, specific IgE and EAR were significantly attenuated following the second TMA challenge. LAR remained robust at 18-24 months and was not significantly different from the response on day 35. CONCLUSIONS: These results demonstrate the persistence of chemical sensitization and further suggest that IgE is not essential for LAR. PMID- 19549026 TI - Basophil activation tests for the diagnosis of food allergy in children. AB - BACKGROUND: Positive skin prick tests (SPT) for food allergens and specific IgE (sIgE) in serum indicate sensitization but do not enable distinction between sensitized but tolerant and clinically allergic patients. OBJECTIVE: Herein, we evaluate the clinical relevance of basophil activation tests (BATs) for peanut or egg allergy diagnosis. METHODS: Thirty-two peanut-allergic, 14 peanut-sensitized (sIgE(+) and/or SPT(+) to peanuts) but tolerant children and 29 controls with no history of an adverse reaction to peanuts were included. Similarly, 31 egg allergic, 14 egg-sensitized children (sIgE(+) and/or SPT(+) to egg white) and 22 controls were studied. Flow cytometric analysis of CD63 expression or CD203c upregulation on basophils and the production of leukotrienes (LT) were performed in response to an in vitro crude peanut extract or ovalbumin (OVA) challenge. RESULTS: After in vitro peanut challenge, the basophils from peanut-allergic children showed significantly higher levels of activation than those from controls (P<0.001). After OVA challenge, a similar distinction (P<0.001) was observed between egg-allergics and controls. Interestingly, the majority of egg- or peanut-sensitized children failed to activate basophils, respectively, in response to OVA and peanut challenge. The sensitivity of the CD63, CD203c and LT assay was 86.7%, 89.5% and 76.0% with a specificity of 94.1%, 97.1% and 94.6% for peanut allergy diagnosis. The corresponding performances of BATs applied to egg allergy diagnosis were 88.9%, 62.5% and 77.8% for the sensitivity and 100%, 96.4% and 96.4% for the specificity. CONCLUSION: Neither conventional tests nor BATs are sensitive and specific enough to predict food allergy accurately. However, BATs may helpfully complete conventional tests, especially SPT, allowing improved discrimination between allergic and non-allergic individuals. PMID- 19549027 TI - Timothy grass pollen major allergen Phl p 1 activates respiratory epithelial cells by a non-protease mechanism. AB - BACKGROUND: Group 1 allergens from grass pollen (e.g. Phl p 1, the major allergen of timothy grass Phleum pratense) cause IgE reactivity in about 95% of allergic subjects and exist in all grass species. The respiratory epithelium represents a first line of contact of the immune system with airborne allergens, functions as physical barrier and is an important immunological regulation system. OBJECTIVE: The aim of this study was to investigate the interaction of Phl p 1 with human respiratory epithelium to elucidate the contribution of epithelial cells to the development of allergic reactions. METHODS: Purified Phl p 1 was used to stimulate A549 cells and transient transfected HEK293 cells. mRNA level of different mediators were investigated by real-time PCR, release of the mediators was determined by ELISA. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and an ex vivo model of the murine trachea were used to investigate a potential proteolytic activity of Phl p 1. RESULTS: Phl p 1 activates respiratory epithelial cells as measured by induction of IL-6, IL-8 and TGF-beta mRNA and release. Phl p 1, in contrast to Der p 1 from the house dust mite, does not exert proteolytic activity, as investigated by microscopic observation and MTT test. In an ex vivo model of the murine trachea we were able to show that Der p 1, in contrast to Phl p 1, enhances the transportation velocity of particles by the trachea, presumably by ATP released from the injured epithelium. CONCLUSION: We conclude that under physiological conditions Phl p 1 affects tracheal epithelial cells through a non-proteolytic activity. Enhancement of TGF-beta expression induced by Phl p 1 together with the increased release of IL-6 and IL-8 might provide an indirect mechanism through which the allergen may cross the epithelial barrier and attracts immunocompetent cells. PMID- 19549028 TI - Characterization of undigested particulate material following microwave digestion of recycled document papers. AB - Recycled document paper was microwave digested in a solution of HNO(3) and H(2)O(2) prior to analysis by inductively coupled plasma mass spectrometry (ICP MS) to determine the trace elemental concentrations within the paper. Undigested particulate material was observed and subsequently characterized as a mixture of kaolin (clay) and TiO(2) by Fourier transform infrared spectroscopy and X-ray diffraction spectroscopy. The effect of the particulate material on the elemental concentrations was then investigated. Paper samples were completely digested in hydrofluoric acid (HF) and element concentrations determined in the HF and HNO(3)/H(2)O digests were statistically compared using Student's t-test (95% confidence limit). Statistical differences in element concentrations between the two digests were observed for only four elements and there was no evidence of element adsorption by the particulate material. Hence, the HNO(3)/H(2)O(2) digestion proved sufficient to digest paper for ICP-MS analysis, eliminating the need to use the hazardous and corrosive HF matrix. PMID- 19549029 TI - Application of DNA forensic techniques for identifying poached guanacos (Lama guanicoe) in Chilean Patagonia*. AB - Guanaco (Lama guanicoe) is a protected and widely distributed ungulate in South America. A poacher, after killing guanacos in Valle Chacabuco, Chilean Patagonia, transported and stored the meat. Samples were retrieved by local police but the suspect argued that the meat was from a horse. Mitochondrial cytochrome b gene (774 pb), 15 loci microsatellites, and SRY gene were used to identify the species, number of animals and their population origin, and the sex of the animals, respectively. Analysis revealed that the samples came from a female (absence of SRY gene) Patagonian guanaco (assignment probability between 0.0075 and 0.0282), and clearly distinguishing it from sympatric ungulates (E-value = 0). Based on the evidence obtained in the field in addition to forensic data, the suspect was convicted of poaching and illegally carrying fire arms. This is the first report of molecular tools being used in forensic investigations of Chilean wildlife indicating its promising future application in guanaco management and conservation. PMID- 19549030 TI - Reconstructing the sequence of events surrounding body disposition based on color staining of bone*. AB - Literature regarding bone color is limited to determining location of primary and secondary dispositions. This research is the first to use bone color to interpret the sequence of events surrounding body disposition. Two scenarios were compared bones buried and then exposed on the ground surface and bones exposed then buried. Forty juvenile pig humeri with minimal tissue were used in each scenario with an additional 20 controls to determine if decomposing tissue affects bone color. Munsell Color Charts were used to record bone color of surface and 2.5 cm cross-sections. Results reveal five main surface colors attributed to soil, sun, hemolysis, decomposition, and fungi. Fungi on buried bones suggests prior surface exposure. Cross-sections of strictly buried bones are identical to buried then exposed bone, stressing the importance of bone surface analysis. Cross-sectioning may help verify remains have been exposed then buried. Decomposition of excess tissue creates minimal color staining. PMID- 19549031 TI - Complete heart block and preserved LV function: does right ventricular pacing site matter? PMID- 19549032 TI - Which is the optimal therapy to prevent thromboembolism after atrial fibrillation ablation procedures in low stroke risk patients, anticoagulation or antiplatelet therapy? PMID- 19549033 TI - Reduction in atrial flutter ablation time by targeting maximum voltage: results of a prospective randomized clinical trial. AB - BACKGROUND: Radiofrequency ablation (RFA) of the cavotricuspid isthmus (CTI) is an established therapy for typical atrial flutter. Previous studies have demonstrated that the CTI is often composed of discrete muscle bundles, and evidence has suggested that these bundles correlate with high-voltage local electrograms in the tricuspid isthmus. This randomized, multicenter clinical trial was designed to prospectively compare the hypothesis that a maximum voltage guided (MVG) technique targets critical conducting bundles in the isthmus, as reflected by a reduction in ablation requirements compared to the anatomical approach to atrial flutter ablation. METHODS: Bidirectional block was achieved in patients undergoing ablation for typical atrial flutter using 1 of 2 randomly assigned methods. The anatomical approach produced a contiguous line of ablation lesions from the inferior aspect of the tricuspid annulus to the inferior vena cava using a standard method. The MVG technique sequentially targeted the maximum voltage local electrograms in the CTI along a similar line. RESULTS: Sixty-nine patients were randomized, with mean age 63 +/- 10 and 58 (84%) male. Among patients in the anatomic group (n = 34), mean ablation time was 11.2 +/- 7.5 minutes compared to 5.9 +/- 3.3 in the MVG group (n = 35) (P = 0.0026). A mean of 14.2 +/- 9.7 ablation lesions were created in the anatomic group, and 7.9 +/- 4.8 in the MVG group (P = 0.0042). CONCLUSIONS: Ablation for atrial flutter using an MVG technique results in significantly less ablation requirements than the traditional approach, potentially by concentrating ablation lesions on the muscle bundles responsible for transisthmus conduction. PMID- 19549034 TI - Novel use of a vascular plug to anchor an azygous vein ICD lead. AB - We describe the case of a young patient with severe hypertrophic cardiomyopathy and marginal defibrillation thresholds (DFTs) at implant of a standard transvenous implantable cardioverter-defibrillator (ICD) system. The patient subsequently experienced multiple failed ICD shocks during a prolonged episode of spontaneous ventricular tachycardia/fibrillation. Placement of a second single coil shocking lead in the azygous vein resulted in acceptable DFTs, but the new lead migrated superiorly within hours of the procedure. To stabilize the lead position, a vascular plug was placed in the distal azygous vein, and the shocking lead screw was actively fixated to the meshwork of the device. Subsequent testing confirmed both adequate defibrillation and stable lead position. PMID- 19549035 TI - Transcranial measurement of cerebral microembolic signals during endocardial pulmonary vein isolation: comparison of three different ablation techniques. AB - INTRODUCTION: Isolation of the pulmonary veins (PVI) using high ablation energy is an effective treatment for atrial fibrillation (AF) with a success rate of 50 95%; however, postoperative neurological complications still occur in 0.5-10%. In this study the incidence of cerebral microembolic signals (MES) as a risk factor for neurological complications is examined during 3 percutaneous endocardial ablation procedure strategies: segmental PVI using a conventional radiofrequency (RF) ablation catheter, segmental PVI using an irrigated RF tip catheter, and circumferential PVI with a cryoballoon catheter (CB). METHODS AND RESULTS: Thirty patients underwent percutaneous endocardial PVI. Ostial isolation was performed in 10 patients with a conventional 4-mm RF catheter (CRF) and in 10 patients with a 4-mm irrigated RF catheter (IRF). A circumferential PVI was performed in 10 patients with a CB. Transcranial Doppler (TCD) monitoring was used to detect MES in the middle cerebral arteries. The total number of cerebral MES differs significantly among the 3 PVI groups; 3,908 cerebral MES were measured with use of the CRF catheter, 1,404 cerebral MES with use of the IRF catheter, and 935 cerebral MES with use of the CB catheter. CONCLUSION: This study demonstrates a significant difference in cerebral MES during PVI with 3 different ablation procedures. The use of an irrigated RF and a cryoballoon produces significantly fewer cerebral MES than the use of conventional RF for a PVI procedure, suggesting a higher risk for neurologic complications using conventional RF energy during a percutaneous PVI procedure. PMID- 19549036 TI - Genetic modulation of brugada syndrome by a common polymorphism. AB - BACKGROUND: Brugada syndrome predisposes some subjects to ventricular tachyarrhythmias and sudden cardiac death. Mutations in SCN5A gene have been associated with approximately 25% of Brugada syndrome patients. A common variant in SCN5A, H558R has shown to improve sodium channel activity in mutated channels. We studied whether common variant H558R has any clinical implications in the phenotype of Brugada syndrome. METHODS: Our study population consisted of Brugada syndrome subjects 75 with SCN5A mutation and 92 without SCN5A mutation. Their mean age was 39 +/- 15 and 42 +/- 17 years, and 65% and 86% were male, respectively. We measured PR-, QRS-, QTc-intervals from leads II and V2 of the 12 lead ECG. We also evaluated J-point amplitude from lead V2 and R'/S ratio from lead aVR (the "aVR sign"). The H558R (A-->G) genotype was detected with direct sequencing of the SCN5A gene. RESULTS: The AA genotype carriers had longer QRS duration in lead II (P = 0.017) and higher J-point elevation in lead V2 (P = 0.013), higher "aVR sign" (P = 0.005) and a trend toward more subjects with symptoms (P = 0.067) than G allele carriers. None of the results were significant in Brugada syndrome subjects without SCN5A mutation. CONCLUSION: The common variant H558R seems to be a genetic modulator of Brugada syndrome among carriers of a SCN5A mutation, in whom the presence of the less common allele G improves the ECG characteristics and clinical phenotype. PMID- 19549037 TI - Quantification of left ventricular asynchrony throughout the whole cardiac cycle with a computed algorithm: application for optimizing resynchronization therapy. AB - INTRODUCTION: Measurement of left ventricular (LV) asynchrony is usually determined on single time points from spectral tissue Doppler imaging (TDI) scans that are frequently difficult to identify or not representative of the whole cardiac cycle. Our aim was to validate a new asynchrony index that evaluates the motion of the LV walls throughout the whole cardiac cycle. METHODS AND RESULTS: Ten healthy volunteers and 50 patients undergoing cardiac resynchronization therapy (CRT) were studied with TDI. Wall displacement tracings from the septal and lateral LV walls were analyzed. Cross-correlation was calculated and 2 indices were obtained to assess LV asynchrony: the time delay and the superposition index (SI) between wall displacements. These results were compared between healthy volunteers and CRT patients, and between responders and nonresponders to CRT. Also, the optimal interventricular (VV) interval was based upon the best matching level. Volunteers showed lower asynchrony indices (83 +/- 2% SI, 17 +/- 8 ms time delay) as compared with CRT patients (63 +/- 15% SI, 73 +/- 60 ms time delay, P < 0.05). Responders also had more LV dyssynchrony than nonresponders (58 +/- 15% SI and 92 +/- 66 ms vs 68 +/- 12% and 48 +/- 34 ms, P < 0.05). The optimum VV interval selected by the computed algorithm showed an excellent concordance (Kappa = 0.90, P < 0.05) with that determined by other validated methods for optimizing the programming of CRT devices. CONCLUSIONS: This approach allows measurement of LV intraventricular asynchrony throughout the cardiac cycle, being useful to determine the optimum VV interval and to select candidates for CRT. PMID- 19549038 TI - Prevalence, predictors, and prognosis of atrial fibrillation early after pulmonary vein isolation: findings from 3 months of continuous automatic ECG loop recordings. AB - INTRODUCTION: Following pulmonary vein isolation (PVI) for atrial fibrillation (AF), early recurrences are frequent, benign and classified as a part of a "blanking period." This study characterizes early recurrences and determines implications of early AF following PVI. METHODS AND RESULTS: Seventy-two consecutive patients (59.8 +/- 10.7 years, 69% male) were studied following PVI for paroxysmal or persistent AF. Subjects were fitted with an external loop recorder for automatic, continuous detection of AF recurrence for 3 months. AF prevalence was highest 2 weeks after PVI (54%) and declined to an eventual low of 22%. A significant number (488, 34%) of recurrences were asymptomatic; however, all patients with > or =1 AF event had > or =1 symptomatic event. No clear predictor of early recurrence was identified. Forty-seven (65%) patients had at least 1 AF episode, predominantly (39 of 47 patients, 83%) within 2 weeks of PVI. Of the 33 patients who did not experience AF within the first 2 weeks, 85% (28/33) were complete responders (P = 0.03) at 12 months. Recurrence at any time within 3 months was not associated with procedural success or failure. CONCLUSIONS: Early AF recurrence peaks within the first few weeks after PVI, but continues at a lower level until the completion of monitoring. A blanking period of 3 months is justified to identify patients with AF recurrences that do not portend procedure failure. Freedom from AF in the first 2 weeks following ablation significantly predicts long-term AF freedom. PMID- 19549039 TI - Primary pulmonary artery sarcoma: surgical management and differential diagnosis with pulmonary embolism and pulmonary valve stenosis. AB - BACKGROUND AND AIM OF THE STUDY: Primary pulmonary artery sarcomas are rare and lethal tumors that are usually diagnosed during surgery or autopsy. We present six cases of primary pulmonary artery sarcomas and discuss clinical features, differential diagnosis, surgical treatment, and outcome of the tumors. METHODS: Between January 1994 and July 2008, six patients were identified with the disease during operation. Three patients were initially diagnosed with pulmonary valve stenosis, and two patients had a presumptive diagnosis of chronic pulmonary embolism. Two patients had simple or partial tumor resection. Four patients had radical tumor resection and homograft reconstruction of the pulmonary arteries. RESULTS: Histological examinations showed five malignant mesenchymomas and one fibrosarcoma. One patient died of refractory pulmonary hypertension during operation. Two patients died 4 months postoperatively because of brain metastases. Two patients were alive for 3 and 9 months, respectively after the operation with recurrent tumor. One patient is alive even 2 years after resection with no signs of recurrence or metastasis. CONCLUSIONS: Because of similar clinical features, pulmonary artery sarcomas are often confused with other pulmonary vascular obstructive diseases. Computed tomography scanning and gadolinium-enhanced magnetic resonance imaging could be useful methods for differential diagnosis. The prognosis is very poor. The survival time after resection varies from several months to several years depending on the presence of recurrence or metastasis. Early diagnosis and radical surgical resection presents the only opportunity for a potential cure. PMID- 19549040 TI - Relationship between myocardial viability and coronary run-off in jeopardized myocardium. AB - OBJECTIVE: The aim of this study was to evaluate the relationship between coronary run-off and myocardial viability in jeopardized regions. METHOD: We studied 50 patients (40 male, mean age: 55.63 +/- 10.54 years) with coronary artery stenosis >70% and ejection fraction <40% referred for viability study via dobutamine stress echocardiography. The relationship between coronary run-off and viability was evaluated. Good run-off demonstrates good or moderate and no run off means poor or no run-off. RESULTS: In the apical region, 33% of the segments with good antegrade run-off were viable and 67% nonviable. Also, 72% of the segments with no run-off were nonviable and 28% viable. In the midportion region, 70% of the segments with good antegrade run-off were viable and 30% nonviable; 50% of the segments with no run-off were nonviable and 50% viable. In the basal region, 85% of the segments with good antegrade run-off were viable and 15% nonviable; 19% of the segments with no run-off were nonviable and 81% viable. The proportion of the nonviable segments increased significantly from the basal to apical regions either with good (p < 0.001) or no run-off (p = 0.004). From 239 viable segments, 58.6% had antegrade, 15.4% retrograde, and 25.5% no run-off. Of 181 nonviable segments, 44% had antegrade, 34% retrograde, and 34.8% no run-off. CONCLUSION: There was more susceptibility to nonviability in the apical regions despite good run-off, while the basal segments showed more viability in spite of having no run-off. The findings may be helpful for selecting patients with coronary artery disease and left ventricular systolic dysfunction that benefit from revascularization. PMID- 19549042 TI - Treatment of obstructive prosthetic heart valve thrombosis in pregnancy. PMID- 19549041 TI - Lung dysfunction following cardiopulmonary bypass. AB - BACKGROUND: It is well known by now that during open heart surgery many detrimental factors are involved in lung injury. The influence mainly of cardiopulmonary bypass (CPB), anesthesia, hypothermia, operation itself, as well as medication and transfusion, can cause a diffuse injury in the lungs, which most often leads to a postoperative pulmonary edema and abnormal gas exchange. METHODS: We performed an unrestricted search of Pubmed Medline and EMBASE from 1966 through 2008. Clinical, experimental, basic science, and review papers were included. RESULTS: The hypothesis that a systemic inflammatory reaction takes place after the use of CPB, could explain most of these effects influences in the lung. On the other hand, the release of various pro-inflammatory cytokines like TNF-a, IL-1, IL-2, IL-6, IL-8, and endotoxin during CPB can lead to the entrapment of neutrophils in the pulmonary capillaries. Consequently, the following chain of reactions is likely to occur: an endothelial cell swelling, plasma and protein extravasation into the interstitial tissue, release of proteolytic enzymes, congestion of the alveoli with plasma, erythrocytes and inflammatory debris. CONCLUSION: In this review we highlight the possible pathophysiological mechanisms implicated in the observed postoperative lung dysfunction. PMID- 19549043 TI - Heartmate II implantation with right coronary bypass grafting in ischemic cardiomyopathy with "fixed" pulmonary hypertension: treatment strategy to protect right ventricular function. AB - A 49-year-old male patient suffering from end-stage ischemic cardiomyopathy with a left ventricular ejection fraction below 15% was presented to redo coronary artery bypass grafting (CABG). Coronary angiogram demonstrated an occluded left anterior descending artery and occluded right coronary artery, perfused retrogradely from the circumflex artery. Since positron emission tomography did not demonstrate viable left ventricular myocardium except for the basis of the left ventricle, CABG was considered futile. Cardiac transplantation was contra indicated due to pharmacologically unresponsive pulmonary artery hypertension. The patient successfully underwent left ventricular assist device implantation in combination with right coronary artery revascularization. The article reflects the regimen of right ventricular preservation in this patient. PMID- 19549044 TI - Remote ischemic conditioning: evolution of the concept, mechanisms, and clinical application. AB - Remote ischemic conditioning is a novel concept of protection against ischemia reperfusion injury. Brief controlled episodes of intermittent ischemia of the arm or leg may confer a powerful systemic protection against prolonged ischemia in a distant organ. This conditioning phenomenon is clinically applicable and can be performed before--preconditioning, during--perconditioning, or after- postconditioning prolonged distant organ ischemia. The remote ischemic conditioning may have an immense impact on clinical practice in the near future. PMID- 19549045 TI - Repair of posterior left ventricular aneurysm through transatrial approach. AB - Repair of posterior left ventricular ischemic aneurysms implies an extracardiac approach to reshape the ventricular geometry frequently associated with mitral surgery. A transatrial technique was described for lesions following mitral surgery or for subvalvular idiopathic cases. A transmitral approach was used for ischemic or traumatic pseudoaneurysm. We describe a case of postinfarction posterior true aneurysm with associated mitral incompetence. Both lesions were treated through an intracardiac approach. The posterior mitral leaflet was detached posteriorly to close the aneurysm with a patch, and the valve replaced sparing all subvalvular apparatus. This technique seems to be safe and allows to treat both lesions avoiding ventriculotomy. PMID- 19549046 TI - Surgical management of univentricular heart with total anomalous pulmonary venous drainage and intrapulmonary vertical vein. AB - We report a patient with univentricular heart, heterotaxy syndrome with total anomalous pulmonary venous connection, and an intrapulmonary vertical vein draining into the left superior vena cava who underwent a successful bidirectional Glenn shunt without cardiopulmonary bypass. A polytetrafluoroethylene tube interposition graft was used between the superior vena cava and the pulmonary artery. This condition is unusual and physiologic palliation, rather than anatomic correction as described in this report, may be appropriate for managing this condition. PMID- 19549047 TI - Unilateral re-expansion pulmonary edema: a rare complication following one-lung ventilation for minimal invasive mitral valve reconstruction. AB - A rare case of a unilateral re-expansion pulmonary edema following one-lung ventilation for minimally invasive mitral valve reconstruction in cardiopulmonary bypass is described. PMID- 19549048 TI - Ascending thoracic aorta aneurysm surgery and aortic valve repair during pregnancy. AB - Open-heart surgery during pregnancy is challenging because it requires attention to both mother and fetus. A 31-year-old pregnant woman underwent surgery for a large ascending aorta aneurysm threatening rupture in the 12th week and then went on to a cesarean section at 38 weeks, which produced a healthy baby. PMID- 19549049 TI - Brain death and donor heart dysfunction: implications in cardiac transplantation. AB - AIM: In this article, the hemodynamic, humoral, and immunological perturbations following brain death (BD) are going to be discussed in a stepwise manner. MATERIALS AND METHODS: BD produces derangements in cardiac function, through a not-yet-well-explained mechanism. Using literature review, we attempted to delineate the "pathophysiology" involved. RESULTS: A severe a-adrenergic stimulation following catecholamine storm results in conditions such that the pulmonary capillary pressure is massively increased. Furthermore, cytokine up regulation, endothelial expression molecules, and neutrophil infiltration produce tissue damage. The end result reflects myocardial necrosis due to reduction of the calcium ATPase activity that leads to myocyte calcium overload and cell death. CONCLUSIONS: Delineation of the mechanisms responsible for donor heart dysfunction (DHD) would be presented. Furthermore, an attempt would be made to apply this knowledge into the clinical practice in order to increase the suitability of donor hearts for transplantation. PMID- 19549050 TI - Evaluation of left ventricular diastolic function after edge-to-edge mitral valve plasty. AB - BACKGROUND: Edge-to-edge mitral valve plasty technique has been widely used to treat bileaflet prolapse. This procedure anchors the correspondence leaflets to create a double-orifice mitral valve. The original mitral valve anatomy is changed, and the opening of mitral valve is restricted. Little is known whether this procedure affects the left ventricular diastolic function. METHODS: Thirty patients with mitral regurgitation were included in this study. Fifteen with posterior leaflet prolapse received quadrangular resection (group 1), 15 with anterior or bileaflet prolapse underwent edge-to-edge procedure (group 2). Acute hemodynamics was monitored with a Swan-Ganz catheter (Edwards Lifesciences LLC, Irvine, CA, USA). Left ventricular diastolic function was also evaluated with echocardiography in 28 patients with sinus rhythm. The ratio of peak E velocity and A velocity (E/A), the ratio of early diastolic peak flow velocity to early diastolic mitral annular movement velocity (E/Em), and the ratio of early diastolic mitral annular velocity to late diastolic mitral annular velocity (Em/Am) were measured before operation and one week after operation. RESULTS: Mitral valve area and mitral regurgitate grade decreased significantly after operation. There was no significant change in pulmonary artery wedge pressure between two groups and in each group before and after operation. Echocardiography evaluation showed there was no significant difference in E/A, E/Em, and Em/Am before and after operation between two groups and in each group. CONCLUSION: Edge to-edge mitral valve plasty procedure has no significant impairment on left ventricular diastolic function. A double-orifice mitral valve has similar hemodynamic behavior with a physiological valve. PMID- 19549051 TI - Immediate clinical outcome after prolonged periods of brain protection: retrospective comparison of hypothermic circulatory arrest, retrograde, and antegrade perfusion. AB - BACKGROUND AND AIM: We aimed to compare the immediate clinical outcome after different cerebral perfusion methods, and examine the factors affecting the mortality and neurological outcome. METHODS: Between 1993 and 2006, 339 patients underwent proximal aortic operations using a period of cerebral protection. Among these, 161 patients (mean age of 55 +/- 12 years) who required cerebral protection longer than 25 minutes were included in the analysis. Ascending aorta with or without root was replaced in all patients. In addition, total arch replacement was performed in 36 patients. All patients were cooled to rectal temperature of 16 degrees C. Hypothermic circulatory arrest without adjunctive perfusion was used in 48 patients. Retrograde or antegrade cerebral perfusion was added in 94 and 19 patients, respectively. The mean duration of total cerebral protection was 42 +/- 17 minutes. RESULTS: Overall mortality was 15.5% (25/161) and did not differ among the perfusion groups. There was no difference in the incidence of overall neurological events, temporary neurological dysfunction, or major stroke among the groups. Multivariate analysis revealed that transfusion of >3 units of blood (p < 0.03) was an incremental risk factor for mortality. History of hypertension (p < 0.03), coexisting systemic diseases (p < 0.005), and transfusion of >3 units of blood (p < 0.04) were predictors of temporary neurological dysfunction. CONCLUSION: In proximal aortic operations requiring prolonged periods of cerebral protection, the mortality and neurological morbidity are not determined by the type of cerebral protection method only. Factors like hypertension and diabetes may play a role in the development of temporary neurological dysfunction. PMID- 19549052 TI - The utility of online panel surveys versus computer-assisted interviews in obtaining substance-use prevalence estimates in the Netherlands. AB - AIMS: Rather than using the traditional, costly method of personal interviews in a general population sample, substance-use prevalence rates can be derived more conveniently from data collected among members of an online access panel. To examine the utility of this method, we compared the outcomes of an online survey with those obtained with the computer-assisted personal interviews (CAPI) method. DESIGN: Data were gathered from a large sample of online panellists and in a two stage stratified sample of the Dutch population using the CAPI method. SETTING: The Netherlands. Participants The online sample comprised 57 125 Dutch online panellists (15-64 years) of Survey Sampling International LLC (SSI), and the CAPI cohort 7204 respondents (15-64 years). MEASUREMENTS: All participants answered identical questions about their use of alcohol, cannabis, ecstasy, cocaine and performance-enhancing drugs. The CAPI respondents were asked additionally about internet access and online panel membership. Both data sets were weighted statistically according to the distribution of demographic characteristics of the general Dutch population. FINDINGS: Response rates were 35.5% (n = 20 282) for the online panel cohort and 62.7% (n = 4516) for the CAPI cohort. The data showed almost consistently lower substance-use prevalence rates for the CAPI respondents. Although the observed differences could be due to bias in both data sets, coverage and non-response bias were higher in the online panel survey. CONCLUSIONS: Despite its economic advantage, the online panel survey showed stronger non-response and coverage bias than the CAPI survey, leading to less reliable estimates of substance use in the general population. PMID- 19549053 TI - Opioid agonist pharmacotherapy in New South Wales from 1985 to 2006: patient characteristics and patterns and predictors of treatment retention. AB - AIMS: The aims of this study were to: examine the number and characteristics of patients entering and re-entering opioid replacement treatment between 1985 and 2006, to examine select demographic and treatment correlates of leaving treatment between 1985 and 2000, and to compare retention rates in methadone and buprenorphine maintenance treatment from 2001 to 2006. DESIGN: A retrospective cohort study using register data from the Pharmaceutical Drugs of Addiction System. SETTING: Opioid substitution treatment in New South Wales (NSW), Australia. PARTICIPANTS: A total of n = 42 690 individuals prescribed opioid replacement treatment between 1985 and 2006 in NSW. MEASUREMENTS: Client characteristics over time, retention in days in first treatment episode, number of episodes of treatment and proportion switching medication. FINDINGS: Overall, younger individuals were significantly more likely to leave their first treatment episode than older individuals. In 2001-06, after controlling for age, sex and first administration point, the hazard of leaving treatment was 1.9 times for those on buprenorphine relative to those on methadone. Retention in treatment varied somewhat across historical time, with those entering during 1995-2000 more likely to leave at an earlier stage than those who entered before that time. CONCLUSIONS: Retention in treatment appears to fluctuate in inverse proportion to the availability of heroin. Individuals in contemporary treatment are older users with a lengthy treatment history. This study has provided population-level evidence to suggest that retention in methadone and buprenorphine differ in routine clinical practice. Future work might investigate ways in which patient adherence and retention may be improved. PMID- 19549054 TI - Smoking cessation during alcohol treatment: a randomized trial of combination nicotine patch plus nicotine gum. AB - AIMS: The primary aim was to compare the efficacy of smoking cessation treatment using a combination of active nicotine patch plus active nicotine gum versus therapy consisting of active nicotine patch plus placebo gum in a sample of alcohol-dependent tobacco smokers in an early phase of out-patient alcohol treatment. A secondary aim was to determine whether or not there were any carry over effects of combination nicotine replacement on drinking outcomes. DESIGN: Small-scale randomized double-blind placebo-controlled clinical trial with 1-year smoking and drinking outcome assessment. SETTING: Two out-patient substance abuse clinics provided a treatment platform of behavioral alcohol and smoking treatment delivered in 3 months of weekly sessions followed by three monthly booster sessions. PARTICIPANTS: Participants were 96 men and women with a diagnosis of alcohol abuse or dependence and smoking 15 or more cigarettes per day. INTERVENTION: All participants received open-label transdermal nicotine patches and were randomized to receive either 2 mg nicotine gum or placebo gum under double-blind conditions. FINDINGS: Analysis of 1-year follow-up data revealed that patients receiving nicotine patch plus active gum had better smoking outcomes than those receiving patch plus placebo gum on measures of time to smoking relapse and prolonged abstinence at 12 months. Alcohol outcomes were not significantly different across medication conditions. CONCLUSIONS: Results of this study were consistent with results of larger trials of smokers without alcohol problems, showing that combination therapy (nicotine patch plus gum) is more effective than monotherapy (nicotine patch) for smoking cessation. PMID- 19549055 TI - Socio-demographic risk factors for alcohol and drug dependence: the 10-year follow-up of the national comorbidity survey. AB - AIMS: Continued progress in etiological research and prevention science requires more precise information concerning the specific stages at which socio demographic variables are implicated most strongly in transition from initial substance use to dependence. The present study examines prospective associations between socio-demographic variables and the subsequent onset of alcohol and drug dependence using data from the National Comorbidity Survey (NCS) and the NCS Follow-up survey (NCS-2). DESIGN: The NCS was a nationally representative survey of the prevalence and correlates of DSM-III-R mental and substance disorders in the United States carried out in 1990-2002. The NCS-2 re-interviewed a probability subsample of NCS respondents a decade after the baseline survey. Baseline NCS socio-demographic characteristics and substance use history were examined as predictors of the first onset of DSM-IV alcohol and drug dependence in the NCS-2. PARTICIPANTS: A total of 5001 NCS respondents were re-interviewed in the NCS-2 (87.6% of baseline sample). FINDINGS: Aggregate analyses demonstrated significant associations between some baseline socio-demographic variables (young age, low education, non-white ethnicity, occupational status) but not others (sex, number of children, residential area) and the subsequent onset of DSM-IV alcohol or drug dependence. However, conditional models showed that these risk factors were limited to specific stages of baseline use. Moreover, many socio-demographic variables that were not significant in the aggregate analyses were significant predictors of dependence when examined by stage of use. CONCLUSIONS: The findings underscore the potential for socio demographic risk factors to have highly specific associations with different stages of the substance use trajectory. PMID- 19549057 TI - Emphasizing interpersonal factors: an extension of the Witkiewitz and Marlatt relapse model. AB - AIM: Recently, Witkiewitz & Marlatt reformulated the Marlatt & Gordon relapse model to account for current research findings. The present paper aims to extend this model further to incorporate social variables more fully. METHODS: The social-factors and alcohol-relapse literatures were reviewed within the framework of the reformulated relapse model. RESULTS: The literature review found that the number of social network members, investment of the individual in the social network, levels of general and alcohol-specific support available within the social network and specific behaviors of network members all predict drinking outcomes. However, little is known about the mechanisms by which these social variables influence outcomes. The authors postulate that social variables influence outcomes by affecting intra-individual factors central to the reformulated relapse prevention model, including processes (e.g. self-efficacy, outcome expectancies, craving, motivation, negative affective states) and behaviors (e.g. coping and substance use). The authors suggest specific hypotheses and discuss methods that can be used to study the impact of social factors on the intra-individual phenomena that contribute to relapse. CONCLUSION: The proposed extension of the relapse model provides testable hypotheses that may guide future alcohol-relapse research. PMID- 19549056 TI - Adolescent smoking and depression: evidence for self-medication and peer smoking mediation. AB - AIMS: The nature of the relationship between adolescent smoking and depression is unclear and the mechanisms that account for the comorbidity have received little investigation. The present study sought to clarify the temporal precedence for smoking and depression and to determine whether these variables are linked indirectly through peer smoking. PARTICIPANTS: The sample was composed of 1093 adolescents participating in a longitudinal study of the behavioral predictors of smoking adoption. DESIGN AND MEASUREMENTS: In this prospective cohort study, smoking, depression, peer smoking and other covariates were measured annually from mid-adolescence (9th grade; age 14) to late adolescence (12th grade, age 18). FINDINGS: Parallel processes latent growth curve models supported a bidirectional relationship between adolescent smoking and depression, where higher depression symptoms in mid-adolescence (age 14) predicted adolescent smoking progression from mid- to late adolescence (ages 14-18). A significant indirect effect indicated that higher depression symptoms across time predicted an increase in the number of smoking peers, which in turn predicted smoking progression from mid-adolescence to late adolescence. In addition, smoking progression predicted a deceleration of depression symptoms from mid- to late adolescence. A significant indirect effect indicated that greater smoking at baseline predicted a deceleration in the number of smoking peers across time, which predicted a deceleration in depression symptoms from mid-adolescence to late adolescence. CONCLUSIONS: The current study provides the first evidence of bidirectional self-medication processes in the relationship between adolescent smoking and depression and highlights peer smoking as one explanation for the comorbidity. PMID- 19549058 TI - Behavioral intervention to promote smoking cessation and prevent weight gain: a systematic review and meta-analysis. AB - AIMS: The prospect of weight gain discourages many cigarette smokers from quitting. Practice guidelines offer varied advice about managing weight gain after quitting smoking, but no systematic review and meta-analysis have been available. We reviewed evidence to determine whether behavioral weight control intervention compromises smoking cessation attempts, and if it offers an effective way to reduce post-cessation weight gain. METHODS: We identified randomized controlled trials (RCTs) that compared combined smoking treatment and behavioral weight control to smoking treatment alone for adult smokers. English language studies were identified through searches of PubMed, Ovid MEDLINE, CINAHL, EMBASE, PsycINFO and Cochrane Central Register of Controlled Trials. Of 779 articles identified and 35 potentially relevant RCTs screened, 10 met the criteria and were included in the meta-analysis. RESULTS: Patients who received both smoking treatment and weight treatment showed increased abstinence [odds ratio (OR) = 1.29, 95% confidence interval (CI) = 1.01, 1.64] and reduced weight gain (g = -0.30, 95% CI = -0.57, -0.02) in the short term (<3 months) compared with patients who received smoking treatment alone. Differences in abstinence (OR = 1.23, 95% CI = 0.85, 1.79) and weight control (g = -0.17, 95% CI = -0.42, 0.07) were no longer significant in the long term (>6 months). CONCLUSIONS: Findings provide no evidence that combining smoking treatment and behavioral weight control produces any harm and significant evidence of short-term benefit for both abstinence and weight control. However, the absence of long-term enhancement of either smoking cessation or weight control by the time-limited interventions studied to date provides insufficient basis to recommend societal expenditures on weight gain prevention treatment for patients who are quitting smoking. PMID- 19549060 TI - Pain responder analysis: use of area under the curve to enhance interpretation of clinical trial results. AB - Interpretation of results on patient-reported pain outcomes from clinical trials should be meaningful to patients and healthcare providers. This study applied an area-under-the-curve (AUC) analysis to responder profiles in a clinical trial of pregabalin for the treatment of fibromyalgia (FM). Data were from a 14-week, randomized, placebo-controlled trial of pregabalin (300, 450, or 600 mg/day) for the treatment of FM in patients meeting American College of Rheumatology criteria for FM and with a baseline pain score of at least 40 mm on the 100-mm pain visual analogue scale. Pain was evaluated in a daily diary by patients using an 11-point numeric rating scale (0 = no pain, 10 = worst possible pain). Response profiles on pain improvement scores and their differences between pregabalin and placebo were assessed using the AUC (derived using the trapezoidal rule) from the responder curve (vertical axis, proportion of subjects; horizontal axis, minimum percent improvement in pain). The AUC can be interpreted as if all responders were improved by the same percentage equal to the AUC divided by 100. The AUCs (2,100 for placebo, and 2,944, 3,170, and 3,349 for pregabalin 300, 450, and 600 mg, respectively) can be considered as if every responder improved by 21, 29, 31, and 33.5% in the responder's respective treatment group. Pain improvement was significantly better with pregabalin (P < 0.05), with pregabalin responders improving by 8.4% (300 mg/day), 10.7% (450 mg/day), and 12.5% (600 mg/day) more than placebo responders. This novel approach demonstrates that responder profiles can provide an enhanced interpretation of pain outcomes for patient care and symptom management. PMID- 19549059 TI - The Danish Barriers Questionnaire-II: preliminary validation in cancer pain patients. AB - OBJECTIVE: The objective of this study was to examine the psychometric properties of the Danish version of the Barriers Questionnaire-II (DBQ-II). METHODS: The validated Norwegian version of the DBQ-II was translated into Danish. Cancer patients for the study were recruited from specialized pain management facilities. Thirty-three patients responded to the DBQ-II, Hospital Anxiety and Depression Scale, and Brief Pain Inventory pain severity scale. RESULTS: A factor analysis of the DBQ-II resulted in six scales. Scale one, Fatalism, consisted of three items addressing fatalistic beliefs regarding cancer pain management. Scale two, Immune System, consisted of three items addressing the belief that pain medications harm the immune system. Scale three, Monitor, consisted of three items addressing the fear that pain medicine masks changes in one's body. Scale four, Communication, consisted of five items addressing the concern that reports of pain distract the physician from treating the cancer, and the belief that "good" patients do not complain. Scale five, Addiction, consisted of two items addressing the fear of becoming addicted to pain medication. Finally, scale six, Tolerance, consisted of three items addressing the fear of getting tolerant to analgesic effect of pain medicine. Items related to medication side effects were analyzed as separate units. The DBQ-II total had an internal consistency of 0.87. The DBQ-II total score was related to measures of pain relief and anxiety. CONCLUSIONS: The DBQ-II seems to be a reliable and valid measure of the barriers to pain management among Danish cancer patients. PMID- 19549061 TI - Reversal of sleep-disordered breathing with opioid withdrawal. AB - Obstructive sleep apnea, central sleep apnea, sleep related hypoventilation, Biot's or ataxic breathing, and cluster breathing are some of the commonly described sleep disorders in patients who are on long-term opioids. Continuous positive airway pressure that is commonly used to treat obstructive sleep apnea may not be effective in treating sleep-disordered breathing in long-term opioid users, and an adaptive servoventilator (ASV) may be needed. We present a 30-year old woman with excessive daytime sleepiness and sleep-disordered breathing for the past 4 years. Medical history was complicated by chronic osteomyelitis, periorbital abscess, and chronic facial pain requiring methadone for pain control for the last 4 years. In this case, ASV, though effective, was not tolerable due to chronic facial pain, and successful withdrawal of methadone at our pain rehabilitation center resolved the sleep-disordered breathing and improved daytime sleepiness. This is to our knowledge the first case report of resolution of sleep-disordered breathing and improvement in daytime sleepiness after withdrawal of long-term opioid use. PMID- 19549062 TI - Foreign body aspiration in children: field report of a German hospital. AB - BACKGROUND: The aspiration of foreign bodies (FB), especially by small children, is a life-threatening situation and can be fatal. The aim of this survey was to study the types of foreign bodies in the upper airways and digestive tract, and the circumstances leading to the aspiration on the basis of hospital records of the Berlin University Hospital in Germany from 1997-2002. METHODS: We performed a retrospective review of hospital records using a standardized protocol. Foreign body aspiration that occurred in children aged 0-14 were considered for inclusion in the database. During the study period, 78 patients with a diagnosis of FB were included in the database. Forty-five patients were male and 33 were female. The median age was 1. RESULTS: In 89.5% of all cases, the children were under the age of 3. Seventy-five of the 78 patients had a foreign body in the trachea/bronchial trees/lungs based on International Classification of Diseases-9 codes at the time of discharge. At the time the injury occurred, the children had either been eating (41.1%) or playing (50.0%). More than 50% of the children were being supervised by an adult at the time the injury occurred. The foreign bodies (FB) were always extracted by using an endoscopic procedure (n= 43 rigid, n= 6 flexible and n= 29 combination of both methods). Moreover, hospitalization was always required due to an institutional requirement. The most commonly found foreign bodies were seeds, nuts, berries and grains. CONCLUSION: Most of the foreign bodies were found in the bronchial tubes, trachea, and lungs. The extraction method from these areas is rigid and/or flexible bronchoscopy or gastrointestinal endoscopy, a procedure requiring anesthesia. There seems to be no association between the aspirated foreign bodies and other purchased objects or packaging material. The fact that a large fraction of the injuries occur under the supervision of the adults suggests that the number and severity of the injuries could be reduced by educating parents and children. Our experience confirms, therefore, that further research into the behavioral aspects leading to FB injuries is needed. PMID- 19549063 TI - Hypohidrosis-related symptoms in pediatric epileptic patients with topiramate. AB - BACKGROUND: Topiramate is one of the most commonly prescribed newer antiepileptic drugs. However, we have encountered quite a few cases of pediatric epileptic patients on topiramate complaining about the symptoms related to hypohidrosis. The aim of this study was to determine the incidence and define the clinical characteristics of hypohidrosis-related symptoms with topiramate in pediatric patients. METHODS: Data was collected prospectively on 264 patients diagnosed as having epilepsy and treated with topiramate at the Department of Pediatrics, Chonbuk National University Hospital between July 2004 and July 2006. The data were collected by direct interview after at least 3 months had elapsed from the initiation of the medication. RESULTS: The study group was composed of 70 boys and 81 girls, with a mean age of 33.1 +/- 43.2 months. The mean duration of topiramate treatment was 13.4 +/- 15.0 months; 52 patients (34.4%) were treated with topiramate only and 99 patients (65.6%) were on polytherapy including topiramate; 59 out of 151 patients (39.1%) experienced hypohidrosis-related symptoms: such as facial flushing, lethargy, itching sensation, irritability with hyperthermia, heat sensation or heat intolerance. However, there were no patients complaining of hypohidrosis-related symptoms among those who were taking antiepileptic drugs other than topiramate. CONCLUSIONS: Our results suggest that topiramate induces hypohidrosis-related symptoms more often than we expected, especially in pediatric patients. We recommend that pediatric epileptic patients taking topiramate should be warned to avoid hot and humid environments, especially during the hot summer season. PMID- 19549064 TI - Beta lactam allergy in children from two ethnically different populations. AB - BACKGROUND: The effects of ethnicity on beta lactam allergy have not been reported. The Negev Desert in Southern Israel is inhabited by two ethnically distinct populations: Jews and Bedouin Muslims. Approximately 60% of the pediatric population of the area is Jewish. Whereas most Jews live in Westernized urban centers, Bedouins are in the process of transition from semi-nomadic conditions to a sedentary lifestyle and the majority of them now live in towns and villages. We sought to determine the rate and characteristics of physician reported beta lactam allergy in Jewish and Bedouin children. METHODS: The medical records of all children registered in five primary community clinics were reviewed and screened for allergy to beta lactam antibiotics. RESULTS: A total of 26,655 medical records were reviewed: of 11,069 Jewish children and 15,586 Bedouin children. Beta lactam allergy was registered in 344 records (1.3%), and was more frequent in Jewish (n= 226, 2.1%) than in Bedouin children (n= 118, 0.8%, P < 0.0001). Beta lactam allergy was more common in boys in both populations (P < 0.01). The clinical features of the reaction that led to the diagnosis of beta lactam allergy were not significantly different between the two ethnic populations, although the specific antibiotics prescribed and the diagnoses for which they were prescribed were different in the two ethnic populations. CONCLUSIONS: We concluded in this study that pediatric beta lactam allergy was registered more frequently in Jewish than in Bedouin children, and in boys more than in girls. PMID- 19549065 TI - Is the maintenance of homeostatic mitochondrial signaling during stress a physiological role for alternative oxidase? AB - All plants maintain a non-energy-conserving pathway of mitochondrial electron transport referred to as alternative oxidase (AOX) respiration. Here, we briefly review some of the most prevailing themes for the metabolic and physiological roles of this respiratory pathway. Many of these themes relate to the potential of AOX to provide metabolic homeostasis in response to fluctuating cellular conditions, such as is often seen during stress. We then review reverse genetic experiments that have been used to test these hypotheses. To date, such experiments have been limited to just two dicot species and have only targeted one member (a stress-induced member) of the AOX multigene family. Nonetheless, the experiments to date strongly reinforce the idea that AOX respiration is of particular importance during abiotic and biotic stress. Finally, we propose that another core role of AOX may be to modulate the strength of a stress-signaling pathway from the mitochondrion that controls cellular responses to stress. In this way, AOX could be acting to provide a degree of signaling homeostasis from the mitochondrion. This hypothesis may provide explanation for some of the disparate results seen in reverse genetic experiments regarding the impact of AOX on the reactive oxygen network and oxidative damage. PMID- 19549066 TI - Populus from high altitude has more efficient protective mechanisms under water stress than from low-altitude habitats: a study in greenhouse for cuttings. AB - Cuttings of Populus przewalskii and P. cathayana, which originated from high and low altitudes in southwest China, were used to examine the effect of water stress on the morphological, physiological and biochemical traits of plants in a greenhouse for one growing season. The dry mass accumulation and allocation, gas exchanges, extent of peroxidation damage, osmotic adjustment and antioxidative defenses, and amounts of pigments were measured to characterize the differences in peroxidation damage and protective mechanisms of two poplar species that contrast in drought tolerance. Under water stress, poplars showed a series of biochemical adjustments and morphological changes as follows: a decrease in leaf relative water content, gas exchanges, plant growth and dry mass accumulation; an increase in relative allocation to roots; an increase in the osmolyte contents (e.g. total amino acids). Additionally, water deficit induced an increase in peroxidation damage [as indicated by an increase in electrolyte leakage, malondialdehyde (MDA), carbonyl (C = O ) and hydrogen peroxide (H(2) O(2) ) content], enhanced activities or contents of antioxidants (e.g. ascorbate peroxidase, guaiacol peroxidase, glutathione redutase and ascorbic acid) and reduced amounts of leaf pigments (e.g. chlorophyll and carotenoid). Furthermore, there were significant differences in the extent of morphological and biochemical changes between the two poplar species. Compared with P. cathayana, P. przewalskii responded to water stress by allocating relatively more to root dry mass, possessing a higher net photosynthesis rate, and having more efficient protective mechanisms, such as more osmolyte accumulation, stronger antioxidant activities and lower chlorophyll/carotenoid ratio. Thus, P. przewalskii suffered less damage as deduced from lower levels of electrolyte leakage, MDA, C=O and H(2) O(2) content. Therefore, P. przewalskii originating from high altitude could possess more efficient protective mechanisms than P. cathayana, which is from low-altitude habitats. PMID- 19549067 TI - Chilling stress and mitochondrial genome rearrangement in the MSC16 cucumber mutant affect the alternative oxidase and antioxidant defense system to a similar extent. AB - The mosaic MSC16 cucumber (Cucumis sativus L.) mutant, which houses a rearranged mitochondrial genome, has altered respiratory chain activity, with a dysfunctional Complex I, increased external NADH dehydrogenases (ND(ex)) activity, and a higher alternative oxidase (AOX) capacity and AOX protein level. In the present study, changes in oxidative defense metabolism resulting from the respiratory chain dysfunction in the MSC16 mutant were compared with those induced by chilling. Chilling increased the enzymatic and non-enzymatic antioxidant defense systems in the wild-type (WT) but not in MSC16, which displays elevated antioxidant defenses as a result of the mitochondrial mutation. The high AOX capacity and protein level in MSC16 were unchanged as a result of chilling, whereas chilling increased these parameters in WT leaves. In mitochondria isolated from WT plants, superoxide was produced to a similar extent in the matrix and the intermembrane space, but in MSC16 mitochondria superoxide was produced largely within the intermembrane space. Mitochondria isolated from both genotypes after chilling showed increased superoxide production within the intermembrane space. Cytochemical detection revealed an increased abundance of H2O2 in the mitochondrial membrane in mesophyll cells of MSC16 leaves. The mitochondrial mutation also resulted in changes in the antioxidative defense system, including AOX, which were similar to those observed following chilling. The results presented here support the hypothesis that AOX is an effective marker of the cellular reprogramming resulting from stress. Moreover, we propose a role for reactive oxygen species (ROS) generated within the mitochondria in signal transduction. PMID- 19549068 TI - Regulation of respiration when the oxygen availability changes. AB - Oxygen is a vital substrate for plant energy metabolism. Since plants do not have a sophisticated mechanism to deliver oxygen to those sites where it is actually needed, a plant cell has to continuously cope with changes of the oxygen tension within the tissue. The actual internal oxygen concentration will depend on the resistance for oxygen diffusion through the tissue, as well as on the actual respiratory activity. This paper discusses the current state of knowledge on the regulation of respiration by the oxygen availability. Contradicting opinions from the literature on plant respiration are reviewed and commented upon. Also, knowledge about the regulation of respiration in animal mitochondria is included. Apart from changes in glycolytic flux, the role of both the cytochrome-c oxidase (COX) and the alternative oxidase (AOX) in the adaptive response of respiration to changes in the oxygen availability are discussed. One hypothesis is formulated which describes an alternative or additional role for AOX. It is suggested that AOX could play a role in maintaining oxygen homeostasis within the mitochondrion. Because of the relative low affinity for oxygen of AOX as compared to COX, the alternative oxidase will not interfere with COX activity, but AOX activity will reduce the free oxygen concentration, thereby decreasing the production of reactive oxygen species (ROS) inside the mitochondrion. PMID- 19549069 TI - Facial EMG and heart rate responses to emotion-inducing film clips in boys with disruptive behavior disorders. AB - We examined aspects of emotional empathy across different physiological response systems in boys with disruptive behavior disorders (DBD) and normal controls. Heart rate (HR) and electromyographic (EMG) reactivity in zygomaticus major and corrugator supercilii muscles were monitored during sadness-, anger-, or happiness-inducing film clips. Relative to controls, DBD boys showed significantly less HR reduction during sadness, and a smaller increase in corrugator EMG activity both during sadness and anger. No significant group differences emerged in HR and zygomaticus EMG responsivity during happiness. We also examined cardiac activity at rest and found higher resting HR and lower respiratory sinus arrhythmia in DBD boys compared to controls. Findings give evidence for a selective impairment in empathy with sadness and anger (not happiness) among DBD boys who exhibit relatively high levels of anxiety and poor emotional control. PMID- 19549070 TI - Cutaneous lymphoid hyperplasia (pseudolymphoma) in a tattoo after far infrared light. PMID- 19549071 TI - Functional dissection of the alpha-synuclein promoter: transcriptional regulation by ZSCAN21 and ZNF219. AB - Alpha-synuclein (SNCA) is an abundant neuronal protein involved in synaptic neurotransmission. SNCA expression levels have been strongly implicated in Parkinson's disease pathogenesis. We have previously demonstrated that in the PC12 cell line elements in intron 1 may mediate SNCA transcriptional regulation in response to neurotrophins. We have now identified transcription factor (TF) binding sites in intron 1 and the 5'-promoter of SNCA. A binding site for the TF zinc finger and SCAN domain containing (ZSCAN)21 in the 5'-region of intron 1 is required for intron 1 transcriptional activity. Small interfering RNA against ZSCAN21 inhibits activation in the luciferase assay and diminishes SNCA protein levels in naive and neurotrophin-treated PC12 cells and in primary cultured cortical neurons, demonstrating that ZSCAN21 is a novel transcriptional regulator of SNCA in neuronal cells. The 5'-promoter of SNCA has a complex architecture, including multiple binding sites for the TF zinc finger protein (ZNF)219, which functions as both an activator and a repressor. Targeting ZSCAN21 or other TFs controlling SNCA transcriptional activity may provide novel therapeutic avenues not only for Parkinson's disease but also for other synucleopathies. PMID- 19549072 TI - Activation of the histaminergic H3 receptor induces phosphorylation of the Akt/GSK-3 beta pathway in cultured cortical neurons and protects against neurotoxic insults. AB - Stimulation of histamine H(3) receptors (H(3)R) activates G(i/o)-proteins that inhibit adenylyl cyclase and triggers MAPK and phospholipase A(2). In a previous study, we showed that H(3)R-mediated phosphorylation of Akt at Ser473 occurs in primary cultures of rat cortical neurons, but neither the downstream targets nor the function of such activation were explored. In this report we address these questions. Western blotting experiments showed that H(3)R-mediated activation of Akt in cultured rat cortical neurons was inhibited by LY 294004 and U0126, suggesting that it depends on phosphoinositide-3-kinase and mitogen-activated protein kinase kinase. H(3)R activation phosphorylated, hence inactivated, the Akt downstream effector glycogen synthase kinase-3beta, increased the expression of the antiapoptotic protein Bcl-2 and protected cultured rat and mouse cortical neurons from neurotoxic insults in a dose-dependent manner. All these effects were inhibited by the H(3)R antagonist inverse/agonist thioperamide. Mouse cortical cells expressed H(3)R as revealed by immunostaining experiments, and stimulation of H(3)R phoshorylated Akt and decreased caspase 3 activity. Hence, we uncovered a yet unexplored action of the H(3)R that may help understand the impact of H(3)R signaling in the CNS. PMID- 19549073 TI - Parkin deficiency increases the resistance of midbrain neurons and glia to mild proteasome inhibition: the role of autophagy and glutathione homeostasis. AB - Parkin mutations in humans produce parkinsonism whose pathogenesis is related to impaired protein degradation, increased free radicals and abnormal neurotransmitter release. In this study, we have investigated whether partial proteasomal inhibition by epoxomicin, an ubiquitin proteasomal system (UPS) irreversible inhibitor, further aggravates the cellular effects of parkin suppression in midbrain neurons and glia. We observed that parkin null (PK-KO) midbrain neuronal cultures are resistant to epoxomicin-induced cell death. This resistance is due to increased GSH and DJ-1 protein levels in PK-KO mice. The treatment with epoxomicin increases, in wild type (WT) cultures, the pro apoptotic Bax/Bcl-2 ratio, the phosphorylation of tau, and the levels of chaperones heat-shock protein 70 and C-terminal Hsc-interacting protein, but none of these effects took place in epoxomicin-treated PK-KO cultures. Poly ubiquitinated proteins increased more in WT than in PK-KO-treated neuronal cultures. Parkin accumulated in WT neuronal cultures treated with epoxomicin. Markers of autophagy, such as LC3II/I, were increased in naive PK-KO cultures, and further increased after treatment with epoxomicin, implying that the blockade of the proteasome in PK-KO neurons triggers the enhancement of autophagy. The treatment with l-buthionine-S,R-sulfoximine and the inhibition of autophagy, however, reverted the increase resistance to epoxomicin of the PK-KO cultures. We also found that PK-KO glial cells, stressed by growth in defined medium and depleted of GSH, were more susceptible to epoxomicin induced cell death than WT glia treated similarly. This susceptibility was linked to reduced GSH levels and less heat-shock protein 70 response, and to activation of p-serine/threonine kinase protein signaling pathway as well as to increased poly-ubiquitinated proteins. These data suggest that mild UPS inhibition is compensated by other mechanisms in PK-KO midbrain neurons. However the depletion of GSH, as happens in stressed glia, suppresses the protection against UPS inhibition-induced cell death. Furthermore, GSH inhibition regulated differentially UPS activity and in old PK-KO mice, which have depletion of GSH, UPS activity is decreased in comparison with that of old-WT. PMID- 19549075 TI - Methods for computer-aided chemical biology. Part 5: rationalizing the selectivity of cathepsin inhibitors on the basis of molecular fragments and topological feature distributions. AB - We report a fragment-based approach to analyze the target selectivity of active compounds. Sets of inhibitors were studied having different activity and selectivity for cathepsins, a family of therapeutically relevant thiol proteases. A systematic analysis was carried out of molecular fragments and atom environment features and their frequency of occurrence for compounds with different selectivity. Fragments extracted from target-selective compounds and independently derived topological features were matched and selectivity markers were identified. Because there is only little overlap between selectivity and other compound set markers, combinations of selectivity set markers could be utilized to predict the selectivity of new cathepsin inhibitors. PMID- 19549074 TI - Fibroblast growth factor receptor-1 phosphorylation requirement for cardiomyocyte differentiation in murine embryonic stem cells. AB - Fibroblast growth factor receptor-1 (Fgfr1) gene knockout impairs cardiac and haematopoietic development in murine embryonic stem cells (mESC). In FGFR1, tyrosine residues Y653 and Y654 are responsible for its tyrosine kinase (TK) activity whereas phosphorylated Y463 and Y766 represent docking sites for intracellular substrates. Aim of this study was the characterization of FGFR1 signalling requirements necessary for cardiomyocyte differentiation in mESC. To this purpose, fgfr1(-/-) mESC were infected with lentiviral vectors harbouring human wild-type hFGFR1 or the Y653/654F, Y463F and Y766F hFGFR1 mutants. The resulting embryonic stem (ES) cell lines were differentiated as embryoid bodies (EBs) and beating foci formation was evaluated. In order to appraise the presence of cells belonging to cardiovascular and haematopoietic lineages, specific markers were analysed by quantitative PCR, whole mount in situ hybridization and immunofluorescence. Transduction with TK(+) hFGFR1 or the TK(+) Y766F-hFGFR1 mutant rescued cardiomyocyte beating foci formation in fgfr1(-/-) EBs whereas the TK(-) Y653/654F-hFGFR1 mutant and the TK(+) Y463F-hFGFR1 mutant were both ineffective. Analysis of the expression of early and late cardiac markers in differentiating EBs confirmed these observations. At variance with cardiomyocyte differentiation, all the transduced TK(+) FGFR1 forms were able to rescue haematopoietic differentiation in EBs originated by infected fgfr1(-/-) mESC, only the TK(-) Y653/654F-hFGFR1 mutant being ineffective. In keeping with these observations, treatment with different signalling pathway inhibitors indicates that protein kinase C and ERK activation are essential for cardiomyocyte but not for haematopoietic differentiation in EBs generated by fgfr1(+/-) approximately mESC. In conclusion, our results suggest that, although FGFR1 kinase activity is necessary for both cardiac and haematopoietic lineage maturation in mESC, phosphorylation of Y463 in the intracellular domain of the receptor is a specific requirement for cardiomyocyte differentiation. PMID- 19549076 TI - Carbonic anhydrase inhibitors: inhibition of cytosolic carbonic anhydrase isozymes II and VII with simple aromatic sulfonamides and some azo dyes. AB - Several substituted benzenesulfonamides were synthesized by various pathways starting from sulfanilamide. The sulfanilamide diazonium salt was reacted with copper (I) halides, potassium iodide and/or aromatic derivatives, leading to 4 halogeno-, and 4-hydroxy-benzenesulfonamides as well as diazo dyes incorporating sulfamoyl moieties. These sulfonamides were assayed as inhibitors of two physiologically relevant isoforms of the zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1), i.e., the cytosolic CA II (ubiquitous), and CA VII (brain-specific enzyme). Good CA inhibitory activity was detected for some of these derivatives, with inhibition constants (Ki) in the range of 17.5-863 nm against CA II; and 30 4200 nm against CA VII. PMID- 19549077 TI - Continuous positive airway pressure treatment for obstructive sleep apnoea reduces resting heart rate but does not affect dysrhythmias: a randomised controlled trial. AB - Obstructive sleep apnoea (OSA) is associated with cardiovascular morbidity and may precipitate cardiac dysrhythmias. Uncontrolled reports suggest that continuous positive airway pressure (CPAP) may reduce dysrhythmia frequency and resting heart rate. We undertook a randomised controlled trial of therapeutic CPAP and compared with a subtherapeutic control which included an exploration of changes in dysrhythmia frequency and heart rate. Values are expressed as mean (SD). Eighty-three men [49.5 (9.6) years] with moderate-severe OSA [Oxygen Desaturation Index, 41.2 (24.3) dips per hour] underwent 3-channel 24-h electrocardiograms during normal daily activities, before and after 1 month of therapeutic (n = 43) or subtherapeutic (n = 40) CPAP. Recordings were manually analysed for mean heart rate, pauses, bradycardias, supraventricular and ventricular dysrhythmias. The two groups were well matched for age, body mass index, OSA severity, cardiovascular risk factors and history. Supraventricular ectopics and ventricular ectopics were frequently found in 95.2% and 85.5% of patients, respectively. Less common were sinus pauses (42.2%), episodes of bradycardia (12%) and ventricular tachycardias (4.8%). Compared with subtherapeutic control, CPAP reduced mean 24-h heart rate from 83.0 (11.5) to 79.7 (9.8) (P < 0.002) in the CPAP group compared with a non-significant rise (P = 0.18) from 79.0 (10.4) to 79.9 (10.4) in the subtherapeutic group; this was also the case for the day period analysed separately. There was no significant change in the frequencies of dysrhythmias after CPAP. Four weeks of CPAP therapy reduces mean 24-h heart rate possibly due to reduced sympathetic activation but did not result in a significant decrease in dysrhythmia frequency. PMID- 19549078 TI - Redistribution of slow wave activity of sleep during pharmacological treatment of depression with paroxetine but not with nefazodone. AB - It has been suggested that increase in delta sleep ratio (DSR), a marker for the relative distribution of slow wave activity (SWA) over night time, is associated with clinical response to antidepressant treatment. We examined this index and its relationship to rapid eye movement (REM) suppression before and during long term treatment with nefazodone, which does not suppress REM sleep, and paroxetine which does. The effect of serotonin (5-HT(2A)) receptor blockade on the evolution of SWA during treatment was also investigated. In a double-blind, randomised, parallel group, 8-week study in 29 depressed patients, sleep electroencephalograms were performed at home at baseline, on night 3 and 10, and at 8 weeks of treatment with either paroxetine or nefazodone. SWA was automatically analysed and a modified DSR (mDSR) was derived, being the ratio of amount of SWA in the first 90 min of sleep to that in the second plus third 90 min periods. At baseline, the pattern of SWA over night time was similar to other reports of depressed patients. mDSR improved over the course of treatment; there was no difference between remitters and non-remitters but there was a significant drug effect and a significant drug x time effect with paroxetine patients having a much higher mDSR after treatment, regardless of clinical status. SWA and REM during antidepressant treatment appear to be interdependent and neither of them alone is likely to predict response to treatment. Higher mDSR did not predict therapeutic response. 5-HT(2A) blockade by nefazodone does not increase SWA above normal levels. PMID- 19549079 TI - Accounting for the richness of daily activities. AB - Serious consideration is being given to the impact of private behavior and public policies on people's subjective well-being (SWB). A new approach to measuring well-being, the day reconstruction method (DRM), weights the affective component of daily activities by their duration in order to construct temporal aggregates. However, the DRM neglects the potentially important role of thoughts. By adapting this method to include thoughts as well as feelings, we provide perhaps the most comprehensive measure of SWB to date. We show that some activities relatively low in pleasure (e.g., work and time with children) are nonetheless thought of as rewarding and therefore contribute to overall SWB. Such information may be important to policymakers wishing to promote behaviors that are conducive to a broader conception of SWB. PMID- 19549080 TI - Selective visual attention and motivation: the consequences of value learning in an attentional blink task. AB - Learning to associate the probability and value of behavioral outcomes with specific stimuli (value learning) is essential for rational decision making. However, in demanding cognitive conditions, access to learned values might be constrained by limited attentional capacity. We measured recognition of briefly presented faces seen previously in a value-learning task involving monetary wins and losses; the recognition task was performed both with and without constraints on available attention. Regardless of available attention, recognition was substantially enhanced for motivationally salient stimuli (i.e., stimuli highly predictive of outcomes), compared with equally familiar stimuli that had weak or no motivational salience, and this effect was found regardless of valence (win or loss). However, when attention was constrained (because stimuli were presented during an attentional blink, AB), valence determined recognition; win-associated faces showed no AB, but all other faces showed large ABs. Motivational salience acts independently of attention to modulate simple perceptual decisions, but when attention is limited, visual processing is biased in favor of reward-associated stimuli. PMID- 19549081 TI - Prismatic lenses shift time perception. AB - Previous studies have demonstrated the involvement of spatial codes in the representation of time and numbers. We took advantage of a well-known spatial modulation (prismatic adaptation) to test the hypothesis that the representation of time is spatially oriented from left to right, with smaller time intervals being represented to the left of larger time intervals. Healthy subjects performed a time-reproduction task and a time-bisection task, before and after leftward and rightward prismatic adaptation. Results showed that prismatic adaptation inducing a rightward orientation of spatial attention produced an overestimation of time intervals, whereas prismatic adaptation inducing a leftward shift of spatial attention produced an underestimation of time intervals. These findings not only confirm that temporal intervals are represented as horizontally arranged in space, but also reveal that spatial modulation of time processing most likely occurs via cuing of spatial attention, and that spatial attention can influence the spatial coding of quantity in different dimensions. PMID- 19549082 TI - Does bilingualism change native-language reading? Cognate effects in a sentence context. AB - Becoming a bilingual can change a person's cognitive functioning and language processing in a number of ways. This study focused on how knowledge of a second language influences how people read sentences written in their native language. We used the cognate-facilitation effect as a marker of cross-lingual activations in both languages. Cognates (e.g., Dutch-English schip [ship]) and controls were presented in a sentence context, and eye movements were monitored. Results showed faster reading times for cognates than for controls. Thus, this study shows that one of people's most automated skills, reading in one's native language, is changed by the knowledge of a second language. PMID- 19549083 TI - The paradox of received social support: the importance of responsiveness. AB - Although the perception of available support is associated with positive outcomes, the receipt of actual support from close others is often associated with negative outcomes. In fact, support that is "invisible" (not perceived by the support recipient) is associated with better outcomes than "visible" support. To investigate this paradox, we proposed that received support (both visible and invisible) would be beneficial when it was responsive to the recipient's needs. Sixty-seven cohabiting couples participated in a daily-experience study in which they reported on the support they provided and received each day. Results indicated that both visible and invisible support were beneficial (i.e., associated with less sadness and anxiety and with greater relationship quality) only when the support was responsive. These findings suggest that the nature of support is an important determinant of when received support will be beneficial. PMID- 19549084 TI - In silico log P prediction for a large data set with support vector machines, radial basis neural networks and multiple linear regression. AB - Oil/water partition coefficient (log P) is one of the key points for lead compound to be drug. In silico log P models based solely on chemical structures have become an important part of modern drug discovery. Here, we report support vector machines, radial basis function neural networks, and multiple linear regression methods to investigate the correlation between partition coefficient and physico-chemical descriptors for a large data set of compounds. The correlation coefficient r(2) between experimental and predicted log P for training and test sets by support vector machines, radial basis function neural networks, and multiple linear regression is 0.92, 0.90, and 0.88, respectively. The results show that non-linear support vector machines derives statistical models that have better prediction ability than those of radial basis function neural networks and multiple linear regression methods. This indicates that support vector machines can be used as an alternative modeling tool for quantitative structure-property/activity relationships studies. PMID- 19549085 TI - QSPR study on the estimation of solubility of drug-like organic compounds: a case of barbiturates. AB - The paper describes a method for the estimation of solubility (log S) of a series of 45 barbiturates employing 26 molecular descriptors. The molecular descriptors used being distance-based topological indices, information indices, valence connectivity index, shape indices, first-order Randic index. In addition, an indicator parameter was also used. The regression analysis has shown that an R(2) value of 0.885 was obtained in multi-parametric regression analysis. The results are discussed critically using a variety of statistical parameters. The predictive powers of the models were discussed by using the method of cross validation. We observed that results obtained using SPSS and NCSS software are identical. PMID- 19549086 TI - Quantitative structure-activity relationship study on the anti-HIV-1 activity of novel 6-naphthylthio HEPT analogs. AB - The quantitative structure-activity relationship of the novel 6-naphthylthio 1 [(2-hydroxyethoxy)methyl]-6-(phenylthio) thymine derivatives for prediction of anti-human immunodeficiency virus type 1 activity was studied. The suitable set of the molecular descriptors was calculated and the important descriptors using the variable selections of the stepwise multiple linear regression and the genetic algorithm were selected. A comparison between the attained results indicated the superiority of the genetic algorithm over the stepwise multiple regression method in the feature-selection. The predictive quality of the quantitative structure-activity relationship models was tested for an external set of eight compounds, randomly chosen out of 39 compounds. The genetic algorithm-multiple linear regression model with six selected descriptors was obtained. This model, demonstrating high statistical qualities (R(2)(train) = 0.925, Q(2) = 0.872, SE (%) = 1.23, F = 49.338, R(2)(pred) = 0.944), could predict the anti-human immunodeficiency virus type 1 activity of the molecules with a prediction error percentage lower than 10%. The results suggest that electronegativity, the masses, and the atomic van der Waals volumes are the main independent factors contributing to the anti-human immunodeficiency virus type 1 activity of the studied compounds. PMID- 19549087 TI - QSAR Studies on antiepileptic and locomotor in vivo activities of 4,5-diphenyl-1H imidazoles. AB - This paper describes the pharmacological evaluation pertaining to in vivo antiepileptic and locomotor activities, and subsequent QSAR studies on 4,5 diphenyl-1H-imidazole analogues. These two activities on albino mice were determined based on electroshock method and by using actophotometer respectively. Compounds with 4-fluorophenyl, 4-dimethylaminophenyl, 4-hydroxyphenyl and 4 methoxyphenyl substitutions exhibit the highest activity. Compounds with phenyl and 2-nitrophenyl substitutions exhibit the lowest activity in both the cases. Data are divided into training and test/validation sets, the former is used for developing the QSAR and the latter is used for determining the predictive capability of the developed models. The three-parameter model for the antiepileptic activity fits the data well and has a good predictive capability (For training set: R( 2) = 0.77, R( 2)(adj) = 0.72, q( 2) = 0.64, R( 2)(mod)= 0.66, for test set: R( 2) = 0.75). Total polar and solvent-accessible surface areas of the molecule are the descriptors in the model. A three parameter model and a four-parameter model for the locomotor activity fit the data well and have good predictive capability (For training set: R( 2) = 0.8-0.89, R( 2)(adj) = 0.76 0.77, q (2) = 0.7-0.86, R( 2)(mod)= 0.64-0.85, for test set: R( 2) = 0.79-0.9). Molecule shape, solvent-accessible surface area, LUMO and polar surface area are the significant descriptors. The first principal component is reverse similar51% and 50% correlated to locomotor and antiepileptic activities respectively. PMID- 19549088 TI - Prevalence of low sexual desire among women in Britain: associated factors. AB - INTRODUCTION: Lack of sexual interest is a common sexual difficulty. Estimates of the prevalence of lack of sexual interest vary widely, and the evidence with regard to factors associated with lack of interest is not always consistent. AIMS: The aims of this study were to identify factors associated with reporting lack of interest in sex among women, and to explore whether these factors differ according to whether or not help was sought. METHODS: Our data came from the second National Survey of Sexual Attitudes and Lifestyles, a probability survey of 12,110 men and women aged 16-44 years and resident in Britain between 1999 and 2001 (N = 6,942 women). Computer-assisted personal interviewing was used to collect sociodemographic, behavioral, and attitudinal data. MAIN OUTCOME MEASURES: (i) Persistent lack of interest in sex (>or=6 months or longer in the past year); and (ii) seeking help for persistent lack of interest in sex. We examined data for all women, regardless of their partnership status. RESULTS: In this study, 10.7% of women reported lacking interest in sex for a period of 6 months or longer, and of these, 27.9% sought help for this difficulty. Reporting persistent low desire per se (outcome 1), and reporting seeking help for low desire (outcome 2) were associated with not enjoying sex, wanting sex more often, not being "competent" at first intercourse, poor communication about sex with partner, frequency of sex, and attitudes according sex low priority. Increasing age, reporting a birth in the last year, having children under 5 in the house, and reporting no sexual partner in the past year were associated with outcome 1 only. Being married and self-perceived health status were associated with outcome 2 only. CONCLUSION: Identifying the factors associated with seeking help for low sexual interest is useful in understanding risk markers for problematic sexual interest, and in providing useful avenues for therapeutic discussion. PMID- 19549090 TI - The role of anxiety in vaginismus: a case-control study. AB - INTRODUCTION: Previous research suggests that anxiety may play a large role in the symptoms of vaginismus. AIM: We aimed to (i) determine the degree of self reported general anxiety in women with vaginismus; and (ii) establish whether general anxiety is a consequence of the condition or a predisposing factor. MAIN OUTCOME MEASURES: Participants reported state and trait anxiety, five-factor personality scores, history of anxiety disorders, and their perceptions of their symptoms and history. METHODS: We compared responses of 244 self-identified women with vaginismus with a control group of 101 women using an online questionnaire. RESULTS: The women with vaginismus were higher in trait anxiety and neuroticism, and lower in extraversion, than the controls. There was also a trend toward a greater prevalence of diagnosed anxiety disorders in the vaginismus group. Levels of state anxiety were high among the women with vaginismus, particularly when they felt unsupported by their partners or pressured to cure the condition. CONCLUSION: Levels of general anxiety are elevated among women with vaginismus and the data suggest that anxiety-proneness may be a predisposing factor for the condition. We conclude that although vaginismus is a multidimensional condition, it may have common predisposing factors with anxiety disorders. PMID- 19549089 TI - Absorption of testosterone gel 1% (Testim) from three different application sites. AB - INTRODUCTION: A popular treatment choice for male hypogonadism is topical testosterone gel. Two proprietary formulations, Testim Gel 1% (Auxilium Pharmaceuticals, Malvern, PA, USA) and AndroGel 1% (Solvay Pharmaceuticals, Marietta, GA, USA), are available. The recommended Testim application site is limited to the arms/shoulders, whereas AndroGel may be applied to the abdomen, shoulders, and upper arms. AIM: To compare absorption variability when applying Testim to various body sites. MAIN OUTCOME MEASURES: Total testosterone (TT) and calculated free testosterone (CT(free)). METHODS: Hypogonadal men (TT < 300 ng/mL) applied Testim to three distinct anatomical sites for 1 month per site: arms/shoulders (A), chest/abdomen (C), and calves/legs (L). Pretreatment TT and CT(free) were compared with end-of-month measurements. Safety was assessed with prostate-specific antigen (PSA) and hemoglobin (Hb) measurements. RESULTS: Twenty one hypogonadal men (age 56.9 +/- 9.0) naive to prior testosterone therapy and otherwise in good health participated. Three groups of seven applied Testim in the sequence ACL, CLA, and LAC. Overall TT and CT(free) increased significantly over pretreatment levels (P < 0.0001) into the normal range. Application sites differed with regard to TT levels achieved, A > C >or= L (P = 0.011). No significant sequence effects were observed, however, the ACL group achieved the highest levels. CT(free) correlated well with TT in all men (R(2) = 0.87) and by application site (R(2) = 0.91, 0.85, and 0.86 for A, C, L, respectively). Pre- and post-treatment PSAs were similar; mean pretreatment Hb increased from 14.7 +/ 1.47 to 15.5 +/- 1.3 g/dL at month 3. Hemoglobin corrected to normal in four subjects with anemia at enrollment (Hb < 13.5 g/dL). CONCLUSIONS: Testim Gel 1% applied to various anatomical sites increases TT and CT(free) into the normal range; the best levels are achieved with arms/shoulder application. Flexibility in the application site of Testim is possible if TT or CT(free) is monitored to ensure adequate therapeutic levels. Anemia, possibly associated with testosterone deficiency, was an incidental finding in several men and was corrected with topical testosterone replacement. PMID- 19549091 TI - Sexual function after radical hysterectomy for early-stage cervical cancer: is there a difference between laparoscopy and laparotomy? AB - INTRODUCTION: Surgical treatment for cervical cancer is associated with a high rate of late postoperative complications, and in particular with sexual dysfunction. AIM: To evaluate sexual function in women who underwent radical hysterectomy (RH), in comparison with a control group of healthy women, using a validated questionnaire (Female Sexual Function Index [FSFI]). Then we tried to evaluate the possible differences between laparoscopic RH and abdominal RH in terms of their impact on sexuality. METHODS: Consecutive sexually active women, who underwent RH for the treatment of early-stage cervical cancer between 2003 and 2007, were enrolled in this study (cases) and divided into two groups, according to the surgical approach. All women were administered the FSFI. The results of this questionnaire were compared between patients who underwent laparoscopic RH (LPS group) vs. women who underwent laparotomic RH (LPT group). The cases of RH were also compared with a control group of healthy women, who were referred to our outpatient clinic for a routine gynecologic evaluation. MAIN OUTCOME MEASURES: FSFI questionnaire on six domains of female sexuality (desire, arousal, lubrication, orgasm, satisfaction, pain). RESULTS: A total of 38 patients were included. We also enrolled 35 women as healthy controls. FSFI score was significantly higher in the healthy controls vs. the cases of RH. In the LPS group, the total score and all the domains of the FSFI were lower in comparison with the healthy controls, whereas three of the six domains (arousal, lubrication, orgasm) and the total score of FSFI were lower in the LPT group if compared with the controls. There were no significant differences between LPS and LPT group. CONCLUSIONS: RH worsens sexual function, regardless of the type of surgical approach. In our experience, laparoscopy did not show any benefit on women's sexuality over the abdominal surgery for cervical cancer. PMID- 19549093 TI - Prevalence and extent of calcification over aorta, coronary and carotid arteries in patients with rheumatoid arthritis. AB - OBJECTIVE: To evaluate the prevalence and pattern of arterial calcification in patients with rheumatoid arthritis (RA). BACKGROUND: Patients with RA are prone to premature atherosclerosis; nonetheless the prevalence and extent of atherosclerosis in different vascular beds and their relationship to each other remain unknown. METHODS: We studied the distribution and extent of arterial calcification in 85 RA patients and 85 age-and sex-matched controls. Arterial calcification as determined by calcium score (CS) were measured using multi detector computed tomography in thoracic aorta, coronary and carotid arteries. RESULTS: Compared with controls, RA patients had a significantly higher average CS and prevalence of CS > 0 in aorta, coronary and carotid arteries and overall arteries (all P < 0.05). After adjusting for age and sex, RA patients had a significantly higher relative risk of developing calcification in the aorta [Odds Ratio (OR) = 19.5, 95% Confidence Interval (CI): 8.0-47.6], followed by the carotid arteries (OR = 5.7, 95% CI:1.7-18.7) and coronary arteries (OR = 5.0, 95% CI:2.2-11.1) compared with controls (all P < 0.01). Amongst RA patients aged >60, 90% had diffuse arterial calcification, especially over the thoracic aorta, compared with 55% of controls who had arterial calcification clustered in the coronary arteries (P < 0.05). RA patients with total CS > 0 were older with a higher urea level and C-reactive protein than those without arterial calcification, no factor was found to be independently predictive for arterial calcification (all P > 0.05). CONCLUSIONS: Our results demonstrated that RA patients had earlier onset, more diffuse arterial calcification over multiple vascular beds and more preferential involvement of thoracic aorta, rather than coronary artery when compared with control. PMID- 19549092 TI - Rituximab in auto-immune haemolytic anaemia and immune thrombocytopenic purpura: a Belgian retrospective multicentric study. AB - OBJECTIVES: For better characterizing the effect of anti-CD20 therapy, we analysed the use of rituximab in Belgian patients experiencing auto-immune haemolytic anaemia (AIHA) and immune thrombocytopenic purpura (ITP). DESIGN: We performed a retrospective multicentric analysis of patients with AIHA and ITP treated with rituximab in Belgium. SETTING: Haematological departments were invited to fill in a questionnaire about patient and disease characteristics. SUBJECTS: All patients with AIHA and ITP, both primary and secondary to other diseases, who received one or more courses of rituximab during their disease course were included. Sixty-eight courses of rituximab in 53 patients with AIHA and 43 courses in 40 patients with ITP were analyzed. INTERVENTION: Response rates, duration of response and factors predictive for response were assessed. RESULTS: All patients were given rituximab after failing at least one previous line of treatment, including splenectomy in 19% and 72.5% of AIHA-patients and ITP-patients respectively. Overall response rates were 79.2% in AIHA and 70% in ITP, with a median follow-up since first rituximab administration of 15 months (range 0.5-62) in AIHA and 11 months (range 0-74) in ITP. Progression free survival at 1 and 2 years were 72% and 56% in AIHA and 70% and 44% in ITP. In this retrospective analysis we were not able to identify pretreatment characteristics predictive for response to rituximab. Nine patients with AIHA and three patients with ITP were given one or more additional courses of rituximab. Most of these patients, who had responded to a previous course, experienced a new response comparable to the previous one, both in terms of quality and of duration of response. Finally, the outcome of patients who failed to respond to rituximab therapy was poor both in terms of response to subsequent therapy and in terms of survival. CONCLUSIONS: This study confirms that rituximab induces responses in a majority of previously treated patients with AIHA and ITP. Response duration generally exceeds 1 year. Retreatment with rituximab in responding patients is most often successful. The outcome of patients who fail on rituximab is poor. We were not able to identify pretreatment patient characteristics predicting for response. PMID- 19549094 TI - Influence of statin treatment on platelet inhibition by clopidogrel - a randomized comparison of rosuvastatin, atorvastatin and simvastatin co-treatment. AB - OBJECTIVES: Possible interactions between clopidogrel and atorvastatin, simvastatin or rosuvastatin (a 'non-CYP3A4' metabolized statin) were investigated in a randomized prospective study using sensitive and specific ex vivo platelet function tests. METHODS: Patients with coronary artery disease participating in a double-blind study comparing lipid-lowering effects of atorvastatin (20-80 mg OD; n = 22) and rosuvastatin (10-40 mg OD; n = 24) were studied before and after 2 weeks treatment with clopidogrel 75 mg OD after completed statin dose titration. In addition, 23 patients were randomized to open-label simvastatin 40 mg OD. RESULTS: Clopidogrel inhibited 10 mumol L(-1) ADP-induced platelet aggregation by 40 +/- 27%, 57 +/- 28% and 51 +/- 29%, respectively, in patients on rosuvastatin, atorvastatin and simvastatin treatment. The other platelet tests yielded similar results. No dose-dependent effects of rosuvastatin or atorvastatin co-treatment on clopidogrel efficacy were observed. CONCLUSIONS: Treatment with CYP3A4 metabolized statins, atorvastatin or simvastatin, did not attenuate the platelet inhibitory effect of clopidogrel maintenance treatment compared with the non CYP3A4 metabolized, rosuvastatin. PMID- 19549095 TI - Persistent parvovirus B19 infection detected by specific CD4+ T-cell responses in a patient with hepatitis and polyarthritis. AB - We, here, report the case of a parvovirus B19 infection in an immunocompetent male patient presenting with acute hepatitis and polyarthritis. To follow the course of infection, we used a previously established enzyme-linked immunosorbent spot assay (ELISPOT) technique to detect CD4+ T cells specific for viral proteins. Even though symptoms of arthritis and hepatitis resolved in the immunocompetent individual within a few weeks, viral DNA in serum and CD4+ T cells specific for the viral protein VP1 unique region were still detectable more than 6 month after the onset of symptoms, thus pointing to a persistent state of infection. On the basis of this observation, we hypothesize that the intensity of liver involvement correlates with the likelihood of developing persistent parvovirus B19 infection. The described ELISPOT technique to detect virus specific CD4+ T cells provides an excellent tool to analyse the state of parvovirus B19 infection for future studies to test this hypothesis. PMID- 19549096 TI - Daytime gating in the Syrian hamster pineal gland. AB - Melatonin synthesis in rodents is tightly regulated at the transcriptional level by stimulatory and inhibitory transcription factors. Among them, phosphorylated cAMP-related element binding protein (pCREB) and inducible cAMP early repressor (ICER), a strong inhibitor of cAMP-related element-driven genes, have an antagonistic action in activating/inhibiting the transcription of the Aa-nat gene, which is an important enzyme in melatonin synthesis. In the Syrian hamster, a rodent displaying a seasonal control of reproduction, melatonin synthesis is strongly gated to the second part of the night. Indeed, exogenous adrenergic stimulation is unable to stimulate Aa-nat gene transcription and melatonin synthesis during daytime. In the present study, we investigated whether ICER may be the cause of this daytime repression by comparing the dynamic of ICER and the adrenergic regulation of two genes whose expression is rapidly activated by cAMP dependant mechanisms, c-fos and Icer. Adrenergic induction of c-fos and Icer expression was not possible during daytime, except at early day. ICER immunoreactivity was elevated throughout the daily cycle but reached the highest levels at early day, when gene expression can be induced by adrenergic agonists. Additionally, CREB phosphorylation was subjected to the same daily gating with an adrenergic induction occurring in the early but not in the late day. Taken together, our results indicate that the diurnal gating of pineal activity in the Syrian hamster is not caused by the repressor ICER and that it may occur at the level of noradrenergic receptor signalling. PMID- 19549097 TI - Melanocytes out-foxed. PMID- 19549098 TI - The ImmunoCAP Rapid Wheeze/Rhinitis Child test is useful in the initial allergy diagnosis of children with respiratory symptoms. AB - Recurrent upper or lower respiratory symptoms, possibly allergy-related, are very frequent in childhood. It is therefore important that physicians involved in the primary care of these children have an accurate initial diagnostic tool available. In this study, we investigated the value of an in vitro diagnostic device testing 10 common allergens, the ImmunoCAP Rapid Wheeze/Rhinitis Child, for the primary evaluation of allergy. Children with non-infectious upper or lower respiratory symptoms possibly related to allergy were recruited in the primary health care setting of private practices of physician trained in immunology/allergology. The investigators carried out their usual diagnostic work up including IgE tests, and the ImmunoCAP Rapid test was performed with capillary whole blood in a blinded way to the investigator. The investigators' conclusions on major triggering allergens were compared to the ImmunoCAP Rapid test results. In the whole patient population (n = 185), the sensitivity of the ImmunoCAP Rapid test for unveiling allergic disease was 92% (95% CI: 86-96%) and the specificity 97% (95% CI: 86-100%). Current guidelines for allergy diagnosis suggest screening children with recurrent, moderate/severe diseases for allergies. For children with asthma falling into these categories, sensitivity was 100% (95% CI: 88-100%) and specificity 100% (95% CI: 69-100%); for children with moderate and severe rhinitis sensitivity was 93% (95% CI: 86-97%) and the specificity 100% (95% CI: 79-100%). The ImmunoCAP Rapid test is an accurate test, in particular with regard to high specificity, for diagnosing allergy in children with recurrent respiratory diseases in primary care settings. PMID- 19549099 TI - Complications of presumed ocular tuberculosis. AB - PURPOSE: To determine the effect of steroid treatment on visual outcome and ocular complications in patients with presumed ocular tuberculosis. METHODS: Retrospective review of patients with presumptive ocular tuberculosis. The clinical diagnosis was made based on ocular findings, positive purified protein derivative (PPD) testing of more than 15 mm induration, exclusion of other causes of uveitis and positive ocular response to anti-tuberculous therapy (ATT) within 4 weeks. Group 1 included patients who had received oral prednisone or subtenon injection of triamcinolone acetonide prior to ATT. Group 2 included patients who did not receive corticosteroid therapy prior to administration of ATT. RESULTS: Among 500 consecutive new cases of uveitis encountered in 1997-2007 there were 49 (10%) patients with presumed ocular tuberculosis. These comprised 28 (57%) male and 21 (43%) female patients with a mean age of 45 years (range 12-76 years). Four (20%) patients in group 1 had initial visual acuity of 20/40 or better, in comparison to eight (28%) patients in group 2. At 1-year follow-up, six (30%) patients in group 1 had a visual acuity of 20/40 or better compared with 20 (69%) patients in group 2 (p = 0.007). Of 20 eyes (26%) in group 1 that had visual acuity of < 20/50 at 1-year follow up, 14 (70%) eyes developed severe chorioretinal lesion (p = 0.019). CONCLUSION: Early administration of corticosteroids without anti-tuberculous therapy in presumed ocular tuberculosis may lead to poor visual outcome compared with patients who did not receive corticosteroids prior to presentation. Furthermore, the severity of chorioretinitis lesion in the group of patients given corticosteroid prior to ATT may account for the poor visual outcome. PMID- 19549100 TI - Efficacy and safety of one intravitreal injection of bevacizumab in diabetic macular oedema. PMID- 19549101 TI - Bilateral giant tear-associated retinal detachment following Artisan phakic intraocular lens implantation for correction of moderate myopia. PMID- 19549103 TI - The inhibitory effect of different concentrations of topical bevacizumab on corneal neovascularization. AB - PURPOSE: This study aimed to evaluate the effects of different concentrations of topically administered bevacizumab (Avastin) on experimental corneal neovascularization (NV) in rats. METHODS: Corneal NV was induced by chemical cauterization with silver nitrate sticks applied to the centre of the corneas of 37 Wistar rats. The rats were then randomized to four topical treatment groups: group 1 (n = 10) received 4 mg/ml bevacizumab; group 2 (n = 9) received 2 mg/ml bevacizumab; group 3 (n = 10) received 1 mg/ml bevacizumab, and group 4 (n=8) represented a control group and received saline. All drops were initiated immediately after cauterization and applied twice per day for 7 days. Corneal NV was assessed 8 days after cauterization in a masked fashion, both qualitatively by clinical evaluation and quantitatively by blood vessel count in photographs of histological sections. RESULTS: On clinical evaluation, groups 1 and 2 showed significantly less NV compared with the saline-treated control group (p = 0.006 and p = 0.024, respectively). Histopathological evaluation showed that only group 1 differed significantly from controls (5% significance level) and normal corneal epithelium was seen in all groups. CONCLUSIONS: Topically administered bevacizumab at a concentration of 4 mg/ml significantly reduces corneal NV according to both clinical and histopathological evaluations; lower concentrations were less effective on both parameters. No corneal epitheliopathy was found using these concentrations. PMID- 19549102 TI - Reduced endothelial progenitor cells and brachial artery flow-mediated dilation as evidence of endothelial dysfunction in ocular hypertension and primary open angle glaucoma. AB - PURPOSE: This study aimed to assess vascular endothelial function in patients with ocular hypertension (OHT) or primary open-angle glaucoma (POAG) by measuring: (a) endothelium-dependent flow-mediated vasodilation (FMD) of the brachial artery, and (b) circulating endothelial progenitor cells, which are believed to support the integrity of the vascular endothelium. METHODS: We enrolled 25 patients with OHT, 23 with POAG and 26 control subjects, all of whom were aged < 65 years and had no medical history of cardiovascular disease or cardiovascular risk factors. All subjects underwent a complete ophthalmological examination, biochemistry study, assessment of cardiovascular parameters, brachial artery ultrasound assessment of endothelium-dependent FMD, generic circulating progenitor cell (CPC) and circulating endothelial progenitor cell (EPC) count with the use of flow cytometry. RESULTS: Flow-mediated vasodilation values differed significantly in OHT (4.5 +/- 1.1%; p = 0.021) and POAG (4.0 +/- 0.9%; p = 0.003) patients compared with controls (7.7 +/- 0.8%). The CD34(+) KDR(+) EPC count was markedly lower in OHT (28.0 +/- 5.0; p < 0.001) and POAG (24.3 +/- 3.4; p < 0.001) patients compared with controls (73.1 +/- 8.1). Neither FMD not EPCs differed significantly between OHT and POAG patients. No significant differences in CPC count or cardiovascular parameters were found among OHT or POAG patients and controls. The levels of CD34(+) KDR(+) EPCs were directly correlated (p = 0.043) with FMD, and inversely correlated (p = 0.032) with baseline intraocular pressure in OHT and POAG patients. CONCLUSIONS: Both OHT and POAG patients without cardiovascular risk factors have previously unreported severely reduced circulating EPCs and reduced FMD, both of which are indicators of endothelial dysfunction and increased risk of cardiovascular events. PMID- 19549104 TI - Nucleolar size in choroidal and ciliary body melanomas and corresponding hepatic metastases. AB - PURPOSE: This study aimed to investigate the relationship between hepatic metastasis and the mean diameter of the 10 largest nucleoli (MLN) in uveal melanoma. METHODS: A cross-sectional histopathological analysis of 37 metastases (13 surgical or needle biopsies, 24 autopsies) and corresponding primary choroidal and ciliary body melanomas was conducted, using statistical tests appropriate for paired data. The largest nucleoli were measured from digital photographs of silver-stained sections along a 5-mm-wide linear field. Confounders considered were presence of epithelioid cells and microvascular density (MVD), counted as the number of discrete elements labelled by monoclonal antibody QBEND/10 to the CD34 epitope. RESULTS: Hepatic metastases had more frequent epithelioid cells (p = 0.0047) and a higher MVD (median difference, 7.5 counts/0.313 mm(2) more; p = 0.044) than their corresponding primary tumours. Hepatic metastases, especially in autopsy specimens rather than surgical biopsies, tended to have a smaller MLN (median 3.6 mum) than the corresponding primary tumour (median difference, 0.55 mum; p = 0.066). The MLN in hepatic metastases was not associated with presence of epithelioid cells and MVD. Overall survival after diagnosis of metastasis was comparable whether hepatic metastases had a large or small MLN (p = 0.95), whereas a high MVD tended to be associated with shorter survival (p = 0.096) among the 13 patients with known survival. CONCLUSIONS: The results suggest that MLN is not a useful marker for assessing prognosis after diagnosis of hepatic metastasis from uveal melanoma. PMID- 19549105 TI - Macular schisis and detachment secondary to large optic nerve head cup: a newly recognized syndrome amenable to vitrectomy. PMID- 19549106 TI - Antifungal activity of bis-azasqualenes, inhibitors of oxidosqualene cyclase. AB - The antifungal activity and in vitro toxicity toward animal cells of two inhibitors of oxidosqualene cyclase, squalene bis-diethylamine (SBD) and squalene bis-diethylmethylammonium iodide (SBDI) were studied. Minimum inhibitory concentration (MIC) against dermatophytes and other fungi involved in cutaneous and systemic infections (12 isolates from seven species) were determined by the broth microdilution method based on the reference documents M38-A and M27-A2 of Clinical and Laboratory Standards Institute (CLSI). Both compounds exerted fungistatic activities, although with different action. SBDI was the more active compound and displayed low MIC values (in the 3.12-12.5 MUg ml(-1) range) against Microsporum canis, Trichophyton mentagrophytes and one isolate of Scopulariopsis brevicaulis, while SBD showed MIC values against these species in the 3.12-25 MUg ml(-1) range. Toxicity was tested on Madin-Darby canine kidney (MDCK) epithelial cells and human microvascular endothelial cells (HMEC). SBDI proved the less toxic compound: it inhibited M. canis, T. mentagrophytes and S. brevicaulis at concentrations below those found toxic for MDCK cells. HMEC were the more sensitive cells. PMID- 19549107 TI - The antifungal properties of chicken egg cystatin against Candida yeast isolates showing different levels of azole resistance. AB - The increasing incidence of fungal infections together with the emergence of strains resistant to currently available antifungal drugs calls for the development of new classes of antimycotics. Naturally occurring antifungal proteins and peptides are of interest because of low toxicity, immunomodulatory potential and mechanisms of action distinct from those of currently available drugs. In this study, the potent antifungal activity of cystatin, affinity purified from chicken egg white (CEWC), against the most frequent human fungal pathogens of the genus Candida was identified and characterised. CEWC inhibited the growth of azole-sensitive Candida albicans isolates with minimal inhibitory concentration (MIC) values ranging from 0.8 to 3.3 micromol l(-1), a potency comparable with those of fluconazole and histatin 5, the antimicrobial peptide of the human saliva. Similarly to histatin 5, CEWC activity was not compromised in azole-resistant isolates overproducing the multidrug efflux transporters Cdr1p and Cdr2p and did not antagonise fluconazole or amphotericin B. CEWC had candidacidal activity, as revealed by the time-kill assay, and, similarly to histatin 5, completely inhibited the growth at supra-MIC concentrations. This was in contrast to the fungistatic effect and trailing growth observed with fluconazole. CEWC inhibited the growth of Candida parapsilosis and Candida tropicalis at similar concentrations, whereas Candida glabrata was more resistant to CEWC. PMID- 19549109 TI - Evidence for divergent natural selection of a Lake Tanganyika cichlid inferred from repeated radiations in body size. AB - Divergent natural selection is thought to play a vital role in speciation, but clear, measurable examples from nature are still few. Among the many possible sources of divergent natural selection, predation pressure may be important because predators are ubiquitous in food webs. Here, we show evidence for divergent natural selection in a Lake Tanganyika cichlid, Telmatochromis temporalis, which uses burrows under stones or empty snail shells as shelters. This species contains normal and dwarf morphs at several localities. The normal morph inhabits rocky shorelines, whereas the dwarf morph invariably inhabits shell beds, where empty snail shells densely cover the lake bottom. Genetic evidence suggested that the dwarf morph evolved independently from the normal morph at two areas, and morphological analysis and evaluation of habitat structure revealed that the body sizes of morphs closely matched the available shelter sizes in their habitats. These findings suggest that the two morphs repeatedly evolved through divergent natural selection associated with the strategy for sheltering from predators. PMID- 19549108 TI - Bathymetric barriers promoting genetic structure in the deepwater demersal fish tusk (Brosme brosme). AB - Population structuring in the North Atlantic deepwater demersal fish tusk (Brosme brosme) was studied with microsatellite DNA analyses. Screening eight samples from across the range of the species for seven loci revealed low but significant genetic heterogeneity (F(ST) = 0.0014). Spatial genetic variability was only weakly related to geographical (Euclidean) distance between study sites or separation of study sites along the path of major ocean currents. Instead, we found a significant effect of habitat, indicated by significant differentiation between relatively closely spaced sites: Rockall, which is surrounded by very deep water (>1000 m), and the Mid-Atlantic Ridge, which is separated from the European slope by a deep ocean basin, were differentiated from relatively homogeneous sites across the Nordic Seas. Limited adult migration across bathymetric barriers in combination with limited intersite exchange of pelagic eggs and larvae due to site-specific circulatory retention or poor survival during drift phases across deep basins may be reducing gene flow. We regard these limitations to gene flow as the most likely mechanisms for the observed population structure in this demersal species. The results underscore the importance of habitat boundaries in marine species. PMID- 19549110 TI - A landscape genetics approach for quantifying the relative influence of historic and contemporary habitat heterogeneity on the genetic connectivity of a rainforest bird. AB - Landscape genetics is an important framework for investigating the influence of spatial pattern on ecological process. Nevertheless, the standard analytic frameworks in landscape genetics have difficulty evaluating hypotheses about spatial processes in dynamic landscapes. We use a predictive hypothesis-driven approach to quantify the relative contribution of historic and contemporary processes to genetic connectivity. By confronting genetic data with models of historic and contemporary landscapes, we identify dispersal processes operating in naturally heterogeneous and human-altered systems. We demonstrate the approach using a case study of microsatellite polymorphism and indirect estimates of gene flow for a rainforest bird, the logrunner (Orthonyx temminckii). Of particular interest was how much information in the genetic data was attributable to processes occurring in the reconstructed historic landscape and contemporary human-modified landscape. A linear mixed model was used to estimate appropriate sampling variance from nonindependent data and information-theoretic model selection provided strength of evidence for alternative hypotheses. The contemporary landscape explained slightly more information in the genetic differentiation data than the historic landscape, and there was considerable evidence for a temporal shift in dispersal pattern. In contrast, migration rates estimated from genealogical information were primarily influenced by contemporary landscape change. We discovered that landscape heterogeneity facilitated gene flow before European settlement, but contemporary deforestation is rapidly becoming the most important barrier to logrunner dispersal. PMID- 19549111 TI - Oral distribution of Candida species and presence of oral lesions in Brazilian leprosy patients under multidrug therapy. AB - OBJECTIVE: The aim of this study was to evaluate the prevalence of Candida spp. and presence of oral lesions in Brazilian leprosy patients under multidrug therapy (MDT). METHODS: Thirty-eight individuals (18 males and 20 females, median age 53 years) clinically and microbiologically diagnosed as leprosy (lepromatous variant), and under MDT for at least 45 days were studied. The control group constituted by 38 healthy individuals (median age 53.5), matched to the test group in relation to age, gender and oral conditions. Oral rinses were collected and the Candida identification was performed by phenotypic tests. The existence of Candida dubliniensis among the isolates was analyzed using a validated multiplex PCR assay. Twenty-nine leprosy patients were examined intra-orally for the presence of lesions. Data were analyzed by z- and Mann-Whitney tests (alpha = 5%). RESULTS: Yeast carriage rate between leprosy patients (65.8%) and controls (47.4%) was similar (P = 0.099), and no significant difference between yeast counts was observed (P = 0.1004). Candida albicans was the most frequently isolated species in both groups. In the leprosy group, Candida tropicalis and Candida parapsilosis were also identified. In the control group, we additionally identified Candida tropicalis, Candida glabrata and Candida kefyr. Candida dubliniensis was not detected. No leprosy-related oral lesion was registered. CONCLUSION: Within the limits of the study, we concluded that Brazilian leprosy patients under MDT showed similar levels of carriage and Candida species distribution in relation to the controls. PMID- 19549112 TI - Targeted drug delivery system for oral cancer therapy using sonoporation. AB - Ultrasound-mediated destruction of microbubbles has been proposed as an innovative non-invasive drug delivery system for cancer therapy. We developed a specific drug delivery system for squamous cell carcinoma that uses sonoporation with the anti-epidermal growth factor receptor (EGFR) antibody. Administration of a low dose of bleomycin (BLM) by sonoporation with the anti-EGFR antibody produced a marked growth inhibition of Ca9-22 cells in vitro. In addition, scanning electron microscopic analysis revealed apparent surface deformation of Ca9-22 cells treated with sonoporation in the presence of the antibody. Interestingly, the population of apoptotic cells was remarkably increased when a low dose of BLM was delivered using sonoporation with the Fab fragment of the anti-EGFR antibody. These findings indicate that sonoporation with the Fab fragment makes it possible to administer drugs into cells more efficiently and specifically, suggesting a novel application for chemotherapy and gene therapy treatments for oral squamous cell carcinoma. PMID- 19549113 TI - Oxidative stress and neurodegeneration in penile ischaemia. AB - OBJECTIVE: To seek markers of oxidative stress and examine neural structural integrity in chronic penile ischaemia using a rabbit model of arteriogenic erectile dysfunction (ED), as the role of ischaemia in penile neuropathy and the oxidative mechanism of neurodegeneration in ED remains unknown. MATERIALS AND METHODS: A rabbit model of atherosclerosis-induced ED was developed by partial balloon de-endothelialization of the iliac arteries. After 10 weeks, intracavernosal blood flow and erectile function in the arteriogenic ED group were compared with age-matched controls. Erectile tissues were processed for analysis of oxidative stress markers and nerve fibre density using enzyme immunoassay and immunohistochemical staining, respectively. Oxidative stress sensitive genes were determined with quantitative real-time polymerase chain reaction. Tissue ultrastructure was examined by transmission electron microscopy. RESULTS: Significant erectile tissue ischaemia, erectile dysfunction, increased levels of oxidative products, and marked nitrotyrosine immunoreactivity was evident in the ED group. Oxidative stress-sensitive genes encoding hypoxia inducible factor-1alpha (HIF-1alpha), superoxide dismutase (SOD), aldose reductase (AR) and nerve growth factor (NGF) were up-regulated in the ischaemic erectile tissue. These changes were associated with collapsed axonal and Schwann cell profiles, neurodegeneration, mitochondrial structural damage, increased caveolae, loss of endothelium, and sporadic vacuolization. CONCLUSIONS: Neuropathy appears to follow the vascular insult in arteriogenic ED. Neural injury in penile ischaemia involves a neurovascular phenomenon mediated by oxidative free radicals. Mitochondrial structural damage and increased HIF-1alpha gene expression may be early signals of oxidative stress and neurodegeneration in ED. Up-regulation of SOD, AR and NGF may be a coordinated defensive reaction to oxidative radicals that seems to fail to prevent neural injury in the ischaemic penis. Our study introduces the concept of oxidative neurodegeneration in the pathophysiology of arteriogenic ED. Therapeutic strategies to protect penile nerves from free radical incursion may enhance the efficacy of surgical and pharmacological interventions in arteriogenic ED. PMID- 19549114 TI - Analysis of association between the 5-HTTLPR and STin2 polymorphisms in the serotonin-transporter gene and clinical response to a selective serotonin reuptake inhibitor (sertraline) in patients with premature ejaculation. AB - OBJECTIVE: To test the hypothesis that polymorphism within the gene-linked polymorphic region (5-HTTLPR) and second intron of SLC6A4 gene (STin2) is associated with selective serotonin-reuptake inhibitors (SSRIs) response in subjects with premature ejaculation (PE). PATIENTS AND METHODS: In all, 246 men with PE were recruited in this study. They were asked to take sertraline 50 mg daily for 2 weeks, and thereafter 100 mg daily, for a 12-week treatment period. Pretreatment evaluation included history and physical examination, intravaginal ejaculatory latency time (IELT), and International Index of Erectile Function (IIEF). The efficacy of treatment was assessed using responses to IIEF, and geometric mean IELT evaluation. 5-HTTLPR was genotyped using polymerase chain reaction techniques. A repeated-measures analysis of variance of geometric mean IELT was done to test a genotype effect on treatment outcome with SSRI (sertraline). RESULTS: Of 227 participants who completed the study, 175 (77.1%) responded to sertraline (IELT >1 min). Overall the patients had a 3.7-fold (95% confidence interval, CI, 1.72-5.46) increase of the geometric mean IELT (P = 0.001). The results showed that responses were significantly better for the L(A)/L(A) genotype of the 5-HTTLPR polymorphism than for S-allele carriers (P = 0.001). The STin2 12/12 genotype was found more often in those responding to sertraline than in those not responding (P = 0.001). The probability of patients responding sufficiently to sertraline with an L(A)/L(A) genotype was highest (odds ratio 4.66, 95% CI, 2.48-6.14). CONCLUSIONS: These findings indicate that the genotype of 5-HTT contributes in unique ways to the variation in the outcome of PE treatment with SSRIs. PMID- 19549115 TI - Do nonspecific deep corticomedullary sutures performed during partial nephrectomy adequately control major vascular and collecting system injury? AB - OBJECTIVE: To critically evaluate the effectiveness of placing nonspecific deep corticomedullary sutures in the setting of major vascular and collecting system injury during laparoscopic partial nephrectomy (LPN). We also aimed to evaluate the incidence of ischaemic injury to the remaining renal remnant because of these sutures, as many laparoscopic centres have adopted this practice. MATERIALS AND METHODS: We performed open PN on eight porcine kidneys. Both the artery and vein were clamped. The ureter was transected and tied around an angiocatheter for evaluating collecting system integrity both before and after corticomedullary suturing. The renal artery was cannulated for angiography before and after the corticomedullary suturing. The rate of bleeding was also assessed before and after corticomedullary suturing. RESULTS: There was marked arterial bleeding and large collecting system injury induced in all kidneys. Two of the eight renal units continued to have significant arterial bleeding after the deep corticomedullary sutures were placed. All of the eight units had at least a small urinary leak after suturing, with three having medium-to-large leaks. In four of the renal units, there were major segmental vessels occluded by the sutures, as detected by angiography. CONCLUSIONS: The practice of placing nonspecific deep corticomedullary sutures, during PN, may not adequately control major vascular and collecting system injury. In addition, segmental vessels supplying remnant renal tissue are often affected; thereby further compromising function because of devascularization. The search for the best technique for LPN continues. PMID- 19549116 TI - National prevalence of urogenital pain and prostatitis-like symptoms in Australian men using the National Institutes of Health Chronic Prostatitis Symptoms Index. AB - STUDY TYPE: Prognosis (cohort). LEVEL OF EVIDENCE: 2a. OBJECTIVE: To provide a summary, using the National Institutes of Health Chronic Prostatitis Symptoms Index (NIH-CPSI), of the prevalence of prostatitis-like symptoms in a population based sample of Australian men. SUBJECTS AND METHODS: Participants were Australian men aged 16-64 years recruited as part of the Australian Longitudinal Study of Health and Relationships: a nationally representative study. In all, 1346 men completed an extensive questionnaire which included the NIH-CPSI. The index identifies six types of urogenital pain, the presence of urinary problems, and effects on quality of life. Men who reported perineal and/or ejaculatory pain or discomfort and a total NIH-CPSI pain score of > or =4 were considered as having prostatitis-like symptoms. RESULTS: Based on a weighted population of 1373 men, some form of urogenital pain was reported by 105 (7.6%) men; with 2.8% of men reporting more than one type of urogenital pain. The mean (range) NIH-CPSI pain score for men reporting pain was 6.2 (5.6-6.8); for all men the mean score was 0.5 (0.4-0.6). About 20% of men (284) were considered to have urinary problems. The mean urinary symptom score for all men was 0.9 (0.9-1.0). The mean total NIH-CPSI score for men reporting pain was 13.3 (12.0-14.7) and for all men it was 2.6 (2.3-2.8). The estimated prevalence of prostatitis-like symptoms was approximately 2%. CONCLUSIONS: Using the NIH-CPSI the estimated prevalence for urogenital pain in Australian men is 8%; an estimated 3% of men experience pain from more than one urogenital location. The estimated prevalence of prostatitis like symptoms in Australian men is 2%. Almost a third of Australian men experiencing urogenital pain or prostatitis-like symptoms would be less than satisfied if this was to be ongoing for the rest of their life. PMID- 19549117 TI - The number of negative pelvic lymph nodes removed does not affect the risk of biochemical failure after radical prostatectomy. AB - OBJECTIVES: To assess patients who had radical prostatectomy (RP) and pelvic lymph node dissection (PLND) for pT2-4 N0M0 prostate cancer, to determine if LN yield affects the risk of biochemical failure (BCF), as the extent of PLND at the time of RP has become increasingly uncertain with the decreasing trend in tumour stage. PATIENTS AND METHODS: We reviewed the Columbia University Urologic Oncology Database for patients with pT2-4 N0M0 prostate cancer treated with RP from 1990 to 2005. Exclusion criteria included <12 months of follow-up, incomplete clinical and pathological data, and neoadjuvant androgen-deprivation therapy (ADT) or immediate adjuvant ADT or external beam radiotherapy. Unadjusted and adjusted models were used to determine the ability of clinical and pathological variables to predict BCF. RESULTS: The final dataset included 964 patients, with a mean age of 60.5 years and median preoperative prostate-specific antigen (PSA) level of 6.2 ng/mL. The median (range) LN yield was 7 (1-42) and the median follow-up 59 (12-190) months. In the unadjusted and adjusted models, preoperative PSA, pathological Gleason score, pathological stage, surgical margin status and year of surgery were significant predictors of BCF. The LN group was not a significant predictor of BCF in both the unadjusted and adjusted model (P = 0.759 and 0.408, respectively). When patients were stratified into high- and low risk groups, LN yield remained an insignificant predictor of BCF. CONCLUSION: A higher LN yield at the time of RP does not increase the chance of cure for patients with pT2-4N0M0 prostate cancer. This lack of a survival advantage holds true for patients with high-risk disease. PMID- 19549118 TI - Neuroanatomy of the male pelvis in respect to radical prostatectomy including three-dimensional visualization. AB - The neuroanatomical structures of the radical prostatectomy (RP) are extensively discussed for their existence, localization and function. Especially structures, e.g. the so-called neurovascular bundle (NVB) that are points of debate in numerous anatomical studies. We review the literature and present our observations in cadaveric specimens, to reconstruct neuroanatomical structures in three dimensions (3D) with the use of appropriate computer applications and produce images of operative fields. We used an internet PubMed survey (http://www.ncbi.nlm.nih.gov) to review recent publications and included back copies of historical neuroanatomical studies from our own library. Our own experimental cadaveric (specimens preserved in Thiel's solution) studies of the autonomic nerve supply of the lower urinary tract were also reviewed. Visualization of the pelvic anatomy and neuroanatomy was done using computer based software packages. No unified terminology for the structures of the NVBs can be presented to date. The innervation of the smooth muscular structures of the urethra and the complex morphology of urethral sphincter remain unclear. Our cadaveric studies showed that nerves are located on the lateral aspect of the prostate in addition to the NVBs described at the dorsolateral side of the prostate. The neuroanatomical investigations of the male pelvis and visualization of the structures in 3D enable the presentation of operative sites as seen intraoperatively. Moreover, dynamic depiction of the pelvic floor is also possible. PMID- 19549119 TI - Adequacy of lymphadenectomy among men undergoing robot-assisted laparoscopic radical prostatectomy. AB - OBJECTIVE: To compare rates of lymph node dissection (LND) and nodal yields between patients treated with open radical retropubic prostatectomy (ORRP) and robot-assisted RRP (RARP) in a contemporary single-institution series. PATIENTS AND METHODS: Data from 1278 consecutive patients (716 ORRP and 562 RARP) from one institution were accrued prospectively in an institutional database, and the data analysed retrospectively. Disease risk was assessed using the Cancer of the Prostate Risk Assessment (CAPRA) score. The likelihood of LND, nodal yield, and likelihood of node positivity were compared between ORRP and RARP. RESULTS: Of patients treated with ORRP and RARP, 47.8% and 31.8% had LND, respectively, with more receiving LND over time in both surgical approaches. Men undergoing LND had a higher disease risk than those not undergoing LND (mean CAPRA score 4.3 vs 2.1, P < 0.01), and there was no difference in risk between those undergoing ORRP or RARP (mean CAPRA score 3.0 vs 2.9, P = 0.29). The mean (sd) nodal yield was 14.4 (8.7) for ORRP and 9.3 (5.4) for RARP (P < 0.01). Among patients undergoing LND, 5.8% of ORRP and 4.1% of RARP patients had positive nodes (P < 0.01). CONCLUSIONS: The indications for LND and template dissection should be the same regardless of surgical approach. The nodal yield was adequate using both approaches; the yield was higher among ORRP than RARP patients, but the difference was not large, and is less remarkable than the wide variation in yield within each approach. Several factors might explain this variation. PMID- 19549120 TI - Payment by results: financial implications of clinical coding errors in urology. PMID- 19549121 TI - Improved detection and reduced early recurrence of non-muscle-invasive bladder cancer using hexaminolaevulinate fluorescence cystoscopy: results of a multicentre prospective randomized study (PC B305). PMID- 19549122 TI - Drug treatment of urological symptoms: estimating the magnitude of unmet need in a community-based sample. AB - OBJECTIVE: To examine, in a community-based sample, the use of prescription drugs for lower urinary tract symptoms/benign prostatic hyperplasia (LUTS/BPH), overactive bladder, erectile dysfunction, urinary incontinence, and painful bladder syndrome; and to determine whether the use of recommended medications varied by sociodemographics, symptom severity, access to care, and other factors. SUBJECTS AND METHODS: In a cross-sectional analysis of data obtained from 5503 men and women residents participating in the Boston Area Community Health Survey of Boston, MA, urological symptoms were ascertained by in-person interviews conducted during 2002-2005, using validated symptom scales. Medication use in the past 4 weeks was captured using a combination of drug-inventory methods and self report. RESULTS: Compared to the prevalence of symptoms, the prevalence of use of medications for urological conditions was very low among men and women. The highest prevalence of use was among men with moderate-to-severe LUTS/BPH symptoms, where 9.6% used recommended drugs. Use of medications did not vary consistently by race/ethnicity or socioeconomic status, but was often associated with symptom severity. More frequent and more recent use of medical care was also associated with greater use of urological medications. CONCLUSIONS: Only a small proportion of community-dwelling men and women with urological symptoms are receiving recommended effective drug treatments for urological conditions. While not all persons are candidates for drug treatment, our results suggest that there is a substantial unmet need in the general population. PMID- 19549123 TI - Interval from prostate biopsy to robot-assisted radical prostatectomy: effects on perioperative outcomes. AB - OBJECTIVE: To determine whether shorter intervals (<4 and 6 weeks) between prostate biopsy and robot-assisted radical prostatectomy (RARP) have a detrimental effect on perioperative outcomes, as recent studies showed that open RP shortly after prostate biopsy does not adversely influence surgical difficulty or efficacy, but RARP relies solely on visual cues rather than tactile sensation to determine posterior surgical planes of dissection. PATIENTS AND METHODS: A series of 559 patients undergoing RARP from March 2004 to July 2007 was retrospectively reviewed. The interval between prostate biopsy and RARP was determined and patients with intervals of 4 weeks. Patient characteristics and perioperative outcomes were analysed to determine statistically significant differences between the groups. This comparison was then repeated with a 6-week interval, and examined with a multivariate logistic regression analysis. RESULTS: In the 4-week group (509 patients), there was a significantly (P < 0.05) higher rate of complications (18.5% vs 6.9%). In the 6-week group (455 patients) there was a smaller but still significantly higher rate of complications (13.6% vs 6.4%). These results were still significant when controlling for patient and disease characteristics and the 'learning curve'. There was also a significantly higher rate of transfusion in the 6-week group (0.7%). CONCLUSIONS: Our data suggest that RARP should be delayed after prostate biopsy; RARP within 6 weeks of biopsy was associated with a greater risk of complications even when controlling for disease and patient characteristics. PMID- 19549124 TI - Does early prostate-specific antigen doubling time (ePSADT) after radical prostatectomy, calculated using PSA values from the first detectable until the first recurrence value, correlate with standard PSADT? A report from the Shared Equal Access Regional Cancer Hospital Database Group. AB - OBJECTIVE: To determine if prostate-specific antigen doubling time (PSADT), calculated from the first detectable PSA level after radical prostatectomy (RP) to the first PSA level of >or=0.2 ng/mL (early PSADT or ePSADT), correlated with 'standard' PSADT (henceforth PSADT) calculated using values >or=0.2 ng/mL, as a short PSADT following biochemical recurrence (BCR) after RP portends a poor prognosis and poor response to salvage treatment but this is based upon PSADT calculated using PSA values of >or=0.2 ng/mL. PATIENTS AND METHODS: We used Spearman's correlation to determine the correlation between ePSADT and PSADT among 157 men in the Shared Equal Access Regional Cancer Hospital database who underwent RP between 1988 and 2005 and had a calculable ePSADT and PSADT. We systematically examined ePSADT thresholds and their positive and negative predictive values (PPV and NPV, respectively), to predict aggressive recurrences (PSADT of <9 months). RESULTS: ePSADT was significantly, though poorly, correlated with PSADT (r = 0.30, P < 0.001). ePSADT more accurately predicted PSADT among men with a long ePSADT. Of men with an ePSADT of >or=20 or >or=15 months, the NPV for an aggressive recurrence was 98% and 93%, respectively. However, among men with an ePSADT of <3 months, the PPV for aggressive recurrence was only 39%. CONCLUSIONS: Although ePSADT and PSADT were significantly related, the overall correlation was poor. This was highlighted by the finding that only 39% of men with the shortest ePSADT (<3 months) had a PSADT of <9 months. However, a long ePSADT correlated well with a long PSADT and is thus useful in identifying men at low risk for prostate cancer-specific mortality very early in their BCR. PMID- 19549125 TI - Population-based prostate-specific antigen testing in the UK leads to a stage migration of prostate cancer. AB - OBJECTIVE: To determine, within the UK, the stage and grade of prostate cancers that would be found through population-based prostate specific antigen (PSA) testing and biopsy. SUBJECTS AND METHODS: In the 'Prostate Testing for Cancer and Treatment' trial (ProtecT), men aged 50-69 years were recruited from nine cities in the UK and from randomly selected practices of general practitioners. Those with a PSA level of >3 ng/mL were offered a prostate biopsy. Age, PSA, stage and grade at diagnosis of ProtecT participants with cancer were compared with contemporaneous incident cases aged 50-69 years (age-restricted Cancer Registry cases) registered with the Eastern Cancer Registration and Information Centre (ECRIC). RESULTS: Within ProtecT, 94,427 men agreed to be tested (50% of men contacted), 8807 ( approximately 9%) had a raised PSA level and 2022 (23%) had prostate cancer; 229 ( approximately 12%) had locally advanced (T3 or T4) or metastatic cancers, the rest having clinically localized (T1c or T2) disease. Within ECRIC, 12,661 cancers were recorded over the same period; 3714 were men aged 50-69 years at diagnosis. Men in ProtecT had a lower age distribution and PSA level, and the cancers were of lower stage and grade (P < 0.001 for all comparisons). If population-based PSA testing were introduced in the UK, approximately 2660 men per 100,000 aged 50-69 years would be found to have prostate cancer, compared to current rates of approximately 130 per 100,000. If half of men accepted PSA testing, approximately 160,000 cancers would be found, compared to 30,000 diagnosed each year at present. CONCLUSIONS: Population-based PSA testing resulted in a significant downward stage and grade migration, and most such cancers were of low stage and grade, which could lead to risks of over treatment for some men. PMID- 19549126 TI - Provision of radical pelvic urological surgery in England, and compliance with improving outcomes guidance. AB - OBJECTIVE: To investigate compliance with Improving Outcomes Guidance (IOG) for radical pelvic surgery in England, and explore the pattern of service provision for radical cystectomy (RC) and radical prostatectomy (RP) before and after the introduction of IOG. METHODS: For the period 2000/01-2006/07, all admissions for RC and RP were extracted from Hospital Episode Statistics (HES). At the institutional level, the numbers of RC and RP cases were combined to assess adherence to IOG. The IOG catchment populations for each institution were calculated by linking HES data to census ward population data. The pattern of service provision for RC and RP was independently assessed by assigning institutions into low-, medium- and high-volume groups of roughly equal volumes a priori, based on the ascending order of annual RC or RP rate, respectively. For RC it was also possible to explore the between-institution referral activity for RC by identifying the 'final endoscopic bladder procedure' that occurred immediately before the RC for each patient. This gave an indication of where the diagnosis and decision for RC had been made. RESULTS: The percentage of institutions achieving the recommended IOG minimal case volume of 50 per year increased significantly between 2000/01 and 2006/07 (36% in odds per year, P < 0.001; odds ratio 1.36, 95% confidence interval 1.24-1.50), although absolute numbers remained relatively low (34% in 2006/07). Only one institution had a catchment population greater than the recommended 1 million. The total number of institutions performing RC decreased significantly over the years (P = 0.03), whereas for RP the decrease was not significant (P = 0.6). The decrease reflected a decline in the number of low-volume institutions, both for RC and RP, although this decline was not more than expected by chance. There had been a significant increase in the percentage of patients referred to another provider for their RC, from 5.5% in 2000/01 to 19.6% in 2006/07 (28% rise in odds per year, P < 0.001: odds ratio 1.28, 95% confidence interval 1.23-1.33). CONCLUSION: There was evidence of centralization of radical pelvic urological surgery, although it is only relatively recently that this seems to have taken place with any certainty. The absolute numbers of providers achieving the IOG minimum caseload standard was relatively low. What impact this has had, if any, on the quality of patient care is yet to be fully determined. PMID- 19549128 TI - Impact of nandrolone decanoate on gene expression in endocrine systems related to the adverse effects of anabolic androgenic steroids. AB - Elite athletes, body builders and adolescents misuse anabolic-androgenic steroids (AAS) in order to increase muscle mass or to enhance physical endurance and braveness. The high doses misused are associated with numerous adverse effects. The purpose of this study was to evaluate the impact of chronic supratherapeutic AAS treatment on circulating hormones and gene expression in peripheral tissues related to such adverse effects. Quantitative real-time PCR was used to measure expression levels of in total 37 genes (including peptide hormones, cell membrane receptors, nuclear receptors, steroid synthesising enzymes and other enzymes) in the pituitary, testes, adrenals, adipose tissue, kidneys and liver of male Sprague-Dawley rats after 14-day administration of the AAS nandrolone decanoate, 3 or 15 mg/kg. Plasma glucose and levels of adrenocorticotropic hormone (ACTH), adiponectin, corticosterone, ghrelin, insulin and leptin were also measured. We found several expected effects on the hypothalamic-pituitary-gonadal axis, while the treatment also caused a number of other not previously identified changes in circulating factors and gene transcription levels such as the dose-dependent reduction of the beta(3)-adrenergic receptor in adipose tissue, reduction of both circulating and mRNA levels of adiponectin, up-regulation of both hydroxymethylglutaryl-CoA-reductase, the rate-limiting enzyme in de novo synthesis of cholesterol, and the receptor for ACTH in the adrenals. The results provide evidence for wide ranging effects of AAS on the hypothalamic-pituitary adrenal axis, adipose tissue and substrates of the renal control of blood pressure. PMID- 19549129 TI - Activation tagging of ADR2 conveys a spreading lesion phenotype and resistance to biotrophic pathogens. AB - An Arabidopsis PR1::luciferase (LUC) transgenic line was transformed with activation T-DNA tags and the resulting population screened for dominant gain-of function mutants exhibiting constitutive LUC activity. LUC imaging identified activated disease resistance 2 (adr2), which exhibited slowly spreading lesions in the absence of pathogen challenge. Molecular, genetic and histochemical analysis was employed to characterize this mutant in detail. adr2 plants constitutively expressed defence-related and antioxidant genes. Moreover, this line accrued increased quantities of salicylic acid (SA) and exhibited heightened mitogen-activated protein kinase activity. adr2 plants exhibited increased resistance against numerous biotrophic but not necrotrophic pathogens. The adr2 phenotype resulted from the overexpression of a Toll interleukin receptor (TIR) nucleotide binding site (NBS) leucine rich repeat (LRR) gene (At1g56510). Constitutive PR1 expression was completely abolished in adr2 nahG, adr2 npr1 and adr2 eds1 double mutants. Furthermore, heightened resistance against Hyaloperonospora arabidopsis Noco2 was compromised in adr2 nahG and adr2 eds1 double mutants but not in adr2 npr1, adr2 coi1 or adr2 etr1 plants. These data imply that adr2-mediated resistance operates through an Enhanced Disease Susceptibility (EDS) and SA-dependent defence signalling network which functions independently from COI1 or ETR1. PMID- 19549130 TI - Genetic and physiological basis of adaptive salt tolerance divergence between coastal and inland Mimulus guttatus. AB - Local adaptation is a well-established phenomenon whereby habitat-mediated natural selection drives the differentiation of populations. However, little is known about how specific traits and loci combine to cause local adaptation. Here, we conducted a set of experiments to determine which physiological mechanisms contribute to locally adaptive divergence in salt tolerance between coastal perennial and inland annual ecotypes of Mimulus guttatus. Quantitative trait locus (QTL) mapping was used to discover loci involved in salt spray tolerance and leaf sodium (Na(+)) concentration. To determine whether these QTLs confer fitness in the field, we examined their effects in reciprocal transplant experiments using recombinant inbred lines (RILs). Coastal plants had constitutively higher leaf Na(+) concentrations and greater levels of tissue tolerance, but no difference in osmotic stress tolerance. Three QTLs contributed to salt spray tolerance and two QTLs to leaf Na(+) concentration. All three salt spray tolerance QTLs had a significant fitness effects at the coastal field site but no effects inland. Leaf Na(+) QTLs had no detectable fitness effects in the field. * Physiological results are consistent with adaptation of coastal populations to salt spray and soil salinity. Field results suggest that there may not be trade-offs across habitats for alleles involved in local salt spray adaptations. PMID- 19549131 TI - Shedding light on an extremophile lifestyle through transcriptomics. AB - The tropical intertidal ecosystem is defined by trees - mangroves - which are adapted to an extreme and extremely variable environment. The genetic basis underlying these adaptations is, however, virtually unknown. Based on advances in pyrosequencing, we present here the first transcriptome analysis for plants for which no prior genomic information was available. We selected the mangroves Rhizophora mangle (Rhizophoraceae) and Heritiera littoralis (Malvaceae) as ecologically important extremophiles employing markedly different physiological and life-history strategies for survival and dominance in this extreme environment. For maximal representation of conditional transcripts, mRNA was obtained from a variety of developmental stages, tissues types, and habitats. For each species, a normalized cDNA library of pooled mRNAs was analysed using GSFLX pyrosequencing. A total of 537,635 sequences were assembled de novo and annotated as > 13,000 distinct gene models for each species. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology annotations highlighted remarkable similarities in the mangrove transcriptome profiles, which differed substantially from the model plants Arabidopsis and Populus. Similarities in the two species suggest a unique mangrove lifestyle overarching the effects of transcriptome size, habitat, tissue type, developmental stage, and biogeographic and phylogenetic differences between them. PMID- 19549132 TI - Speciation and distribution of arsenic and localization of nutrients in rice grains. AB - Arsenic (As) contamination of rice grains and the generally low concentration of micronutrients in rice have been recognized as a major concern for human health. Here, we investigated the speciation and localization of As and the distribution of (micro)nutrients in rice grains because these are key factors controlling bioavailability of nutrients and contaminants. Bulk total and speciation analyses using high-pressure liquid chromatography (HPLC)-inductively coupled plasma mass spectrometry (ICP-MS) and X-ray absorption near-edge spectroscopy (XANES) was complemented by spatially resolved microspectroscopic techniques (micro-XANES, micro-X-ray fluorescence (micro-XRF) and particle induced X-ray emission (PIXE)) to investigate both speciation and distribution of As and localization of nutrients in situ. The distribution of As and micronutrients varied between the various parts of the grains (husk, bran and endosperm) and was characterized by element-specific distribution patterns. The speciation of As in bran and endosperm was dominated by As(III)-thiol complexes. The results indicate that the translocation from the maternal to filial tissues may be a bottleneck for As accumulation in the grain. Strong similarities between the distribution of iron (Fe), manganese (Mn) and phosphorus (P) and between zinc (Zn) and sulphur (S) may be indicative of complexation mechanisms in rice grains. PMID- 19549133 TI - Overexpression of EgROP1, a Eucalyptus vascular-expressed Rac-like small GTPase, affects secondary xylem formation in Arabidopsis thaliana. AB - To better understand the genetic control of secondary xylem formation in trees we analysed genes expressed during Eucalyptus xylem development. Using eucalyptus xylem cDNA libraries, we identified EgROP1, a member of the plant ROP family of Rho-like GTPases. These signalling proteins are central regulators of many important processes in plants, but information on their role in xylogenesis is scarce. Quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) confirmed that EgROP1 was preferentially expressed in the cambial zone and differentiating xylem in eucalyptus. Genetic mapping performed in a eucalyptus breeding population established a link between EgROP1 sequence polymorphisms and quantitative trait loci (QTLs) related to lignin profiles and fibre morphology. Overexpression of various forms of EgROP1 in Arabidopsis thaliana altered anisotropic cell growth in transgenic leaves, but most importantly affected vessel element and fibre growth in secondary xylem. Patches of fibre-like cells in the secondary xylem of transgenic plants showed changes in secondary cell wall thickness, lignin and xylan composition. These results suggest a role for EgROP1 in fibre cell morphology and secondary cell wall formation making it a good candidate gene for marker-based selection of eucalyptus trees. PMID- 19549134 TI - Dissecting iron deficiency-induced proton extrusion in Arabidopsis roots. AB - Here, we have analysed the H(+)-ATPase-mediated extrusion of protons across the plasma membrane (PM) of rhizodermic cells, a process that is inducible by iron (Fe) deficiency and thought to serve in the mobilization of sparingly soluble Fe sources. The induction and function of Fe-responsive PM H(+)-ATPases in Arabidopsis roots was investigated by gene expression analysis and by using mutants defective in the expression or function of one of the isogenes. In addition, the expression of the most responsive isogenes was investigated in natural Arabidopsis accessions that have been selected for their in vivo proton extrusion activity. Our data suggest that the rhizosphere acidification in response to Fe deficiency is chiefly mediated by AHA2, while AHA1 functions as a housekeeping isoform. The aha7 knock-out mutant plants showed a reduced frequency of root hairs, suggesting an involvement of AHA7 in the differentiation of rhizodermic cells. Acidification capacity varied among Arabidopsis accessions and was associated with a high induction of AHA2 and IRT1, a high relative growth rate and a shoot-root ratio that was unaffected by the external Fe supply. An effective regulation of the Fe-responsive genes and a stable shoot-root ratio may represent important characteristics for the Fe uptake efficiency. PMID- 19549135 TI - Association among interleukin-6 gene polymorphism, diabetes and periodontitis in a Chinese population. AB - OBJECTIVES: Diabetics significantly increase risk for periodontitis. Interleukin 6 (IL-6) gene polymorphism may play certain roles in the progression of periodontitis with diabetes. The purpose of this study was to assess the association among IL-6 gene polymorphisms, type 2 diabetes mellitus (T2DM) and chronic periodontitis (CP) in a Chinese population. MATERIAL AND METHODS: DNA was obtained from 159 patients with CP, 88 patients with T2DM, 110 patients with CP&T2DM and 135 control subjects. The -174/-572/-597 polymorphisms of IL-6 gene were investigated by restriction fragment length polymorphism of polymerase chain reaction products. The results were further confirmed by sequencing. Significance was set at P < 0.008 after Bonferroni correction. RESULTS: Among four groups, CP&T2DM group showed the lowest IL-6-572 CC genotype and C-allele frequencies (54.5% and 74.1%). In this regard, there were significant differences between CP&T2DM group and the control group [P = 0.006, odds ratio (OR) = 0.475, 95% CI: 0.279-0.808 and P = 0.002, OR = 0.502, 95% CI: 0.319-0.788 respectively]. Logistic regression with adjustment for age, gender, body mass index, smoking and stress showed no significant difference in terms of IL-6-572 genotypes (P = 0.058, OR= 0.523, 95% CI: 0.268-1.022). CONCLUSIONS: The IL-6-572 genotype and allele distributions are unique to subjects with CP&T2DM in a Chinese population. PMID- 19549136 TI - Influence of clinical parameters on the results of 13C-octanoic acid breath tests: examination of different mathematical models in a large patient cohort. AB - It is assumed, although not proven, that 13CO2-excretion following ingestion of 13C-octanoic acid (13C-OA) does not only depend on gastric emptying (GE) but also on absorption and metabolism of 13C-OA and endogenous CO2-production. Our aims were (i) to test the effects of patient characteristics and of diseases that may impair 13C-OA-metabolism on GE parameters. (ii) To compare different GE endpoints. Therefore, we investigated effects of age, gender, BMI and diseases with potential impact on 13C-OA-metabolism (including pancreatic, liver and lung disease, diabetes, IBD) on cumulative 4h-13CO2-excretion (4h-CUM) and T1/2 calculated by non-linear regression model (NL, determined by shape of breath test curve) and generalized linear regression model (GLR, reflects absolute 13CO2 excretion) in 1279 patients and 19 healthy controls who underwent a standardized 13C-OA-breath test. Digestive and metabolic disturbances hardly influenced 4h-CUM or T1/2 calculated by NL or GLR models. In the multivariate linear regression models, 4h-CUM was significantly predicted by diabetes adjusted for age, gender and IBD but influence of these parameters was small (R2 = 0.028, P < 0.0001). T1/2(NL) and 4h-CUM were weakly correlated, even after exclusion of tests with unrealistically high estimates for T1/2(NL) (n = 1095, R(2) = 0.029, P < 0.0001). Conversely, 4h-CUM was closely associated with T(1/2)(GLR) (exponential correlation, R(2) = 0.774, P < 0.00001, n = 1279). We conclude that influences of digestive and metabolic disturbances on 13CO2-excretion following 13C-OA application are generally low. Thus, our findings resolve an important criticism of methods using absolute 13CO2-excretion for evaluation of 13C-OA-breath tests and suggest that such models may correctly identify T1/2 in a mixed patient population. PMID- 19549137 TI - Major disruption of gene expression in hybrids between young sympatric anadromous and resident populations of brook charr (Salvelinus fontinalis Mitchill). AB - Genome-wide analyses of the transcriptome have suggested that male-biased genes are the first targets of genomic incompatibilities (g.i.) in inter-specific hybrids. However, those studies have almost invariably focused on Drosophila species that diverged at least 0.9 Ma, and with sterile male hybrids. Here, we use microarrays to analyse patterns of gene expression in very closely related (divergence <12,000 years), sympatric, but ecologically divergent anadromous and resident populations of brook charr (Salvelinus fontinalis) and their F(1) hybrids. Our results show a dramatic breakdown of gene expression patterns in hybrids compared with their parental relatives. Several disrupted genes are related to energetic metabolism, immune response, osmoregulation and protection against oxidative stress, and none has sex-biased functions. Besides, pure individuals show no expression differences at most of the genes disrupted in hybrids, which may suggest the operation of some form of stabilizing selection. Taken together, these results both confirm the idea that perturbations of regulatory networks represent a significant source of g.i. and support the suggestion that developmental pathways can diverge through time without any manifest change in the phenotypic outcome. While the role of other evolutionary forces (e.g. genetic drift) cannot be ruled out, this study suggests that ecological selective processes may provide the initial driving force behind disruption of gene expression in inter-specific hybrids. PMID- 19549138 TI - Eusociality and the success of the termites: insights from a supertree of dictyopteran families. AB - Sociality in insects may negatively impact on species richness. We tested whether termites have experienced shifts in diversification rates through time. Supertree methods were used to synthesize family-level relationships within termites, cockroaches and mantids. A deep positive shift in diversification rate is found within termites, but not in the cockroaches from which they evolved. The shift is responsible for most of their extant species richness suggesting that eusociality is not necessarily detrimental to species richness, and may sometimes have a positive effect. Mechanistic studies of speciation and extinction in eusocial insects are advocated. PMID- 19549139 TI - Aphid genotypes vary in their response to the presence of fungal endosymbionts in host plants. AB - Genetic variation for fitness-relevant traits may be maintained in natural populations by fitness differences that depend on environmental conditions. For herbivores, plant quality and variation in chemical plant defences can maintain genetic variation in performance. Apart from plant secondary compounds, symbiosis between plants and endosymbiotic fungi (endophytes) can produce herbivore-toxic compounds. We show that there is significant variation among aphid genotypes in response to endophytes by comparing life-history traits of 37 clones of the bird cherry-oat aphid Rhopalosiphum padi feeding on endophyte-free and endophyte infected tall fescue Lolium arundinaceum. Clonal variation for life-history traits was large, and most clones performed better on endophyte-free plants. However, the clones differed in the relative performance across the two environments, resulting in significant genotype x environment interactions for all reproductive traits. These findings suggest that natural variation in prevalence of endophyte infection can contribute to the maintenance of genetic diversity in aphid populations. PMID- 19549140 TI - Adaptive brain size divergence in nine-spined sticklebacks (Pungitius pungitius)? AB - Most studies seeking to provide evolutionary explanations for brain size variability have relied on interspecific comparisons, while intraspecific studies utilizing ecologically divergent populations to this effect are rare. We investigated the brain size and structure of first-generation laboratory-bred nine-spined sticklebacks (Pungitius pungitius) from four geographically and genetically isolated populations originating from markedly different habitats. We found that the relative size of bulbus olfactorius and telencephalon was significantly larger in marine than in pond populations. Significant, but habitat independent population differences were also found in relative brain and cerebellum sizes. The consistent, habitat-specific differences in the relative size of bulbus olfactorius and telencephalon suggest their adaptive reduction in response to reduced (biotic and abiotic) habitat complexity in pond environments. In general, the results suggest that genetically based brain size and structure differences can evolve relatively rapidly and in repeatable fashion with respect to habitat structure. PMID- 19549141 TI - Phenotypic divergence but not genetic distance predicts assortative mating among species of a cichlid fish radiation. AB - The hypothesis of ecological divergence giving rise to premating isolation in the face of gene flow is controversial. However, this may be an important mechanism to explain the rapid multiplication of species during adaptive radiation following the colonization of a new environment when geographical barriers to gene flow are largely absent but underutilized niche space is abundant. Using cichlid fish, we tested the prediction of ecological speciation that the strength of premating isolation among species is predicted by phenotypic rather than genetic distance. We conducted mate choice experiments between three closely related, sympatric species of a recent radiation in Lake Mweru (Zambia/DRC) that differ in habitat use and phenotype, and a distantly related population from Lake Bangweulu that resembles one of the species in Lake Mweru. We found significant assortative mating among all closely related, sympatric species that differed phenotypically, but none between the distantly related allopatric populations of more similar phenotype. Phenotypic distance between species was a good predictor of the strength of premating isolation, suggesting that assortative mating can evolve rapidly in association with ecological divergence during adaptive radiation. Our data also reveals that distantly related allopatric populations that have not diverged phenotypically, may hybridize when coming into secondary contact, e.g. upon river capture because of diversion of drainage systems. PMID- 19549142 TI - The diversification of mate preferences by natural and sexual selection. AB - The evolution of sexual display traits or preferences for them in response to divergent natural selection will alter sexual selection within populations, yet the role of sexual selection in ecological speciation has received little empirical attention. We evolved 12 populations of Drosophila serrata in a two-way factorial design to investigate the roles of natural and sexual selection in the evolution of female mate preferences for male cuticular hydrocarbons (CHCs). Mate preferences weakened in populations evolving under natural selection alone, implying a cost in the absence of their expression. Comparison of the vectors of linear sexual selection revealed that the populations diverged in the combination of male CHCs that females found most attractive, although this was not significant using a mixed modelling approach. Changes in preference direction tended to evolve when natural and sexual selection were unconstrained, suggesting that both processes may be the key to initial stages of ecological speciation. Determining the generality of this result will require data from various species across a range of novel environments. PMID- 19549143 TI - Testosterone, growth and the evolution of sexual size dimorphism. AB - The integration of macroevolutionary pattern with developmental mechanism presents an outstanding challenge for studies of phenotypic evolution. Here, we use a combination of experimental and comparative data to test whether evolutionary shifts in the direction of sexual size dimorphism (SSD) correspond to underlying changes in the endocrine regulation of growth. First, we combine captive breeding studies with mark-recapture data to show that male-biased SSD develops in the brown anole lizard (Anolis sagrei) because males grow significantly faster than females as juveniles and adults. We then use castration surgeries and testosterone implants to show that castration inhibits, and testosterone stimulates, male growth. We conclude by reviewing published testosterone manipulations in other squamate reptiles in the context of evolutionary patterns in SSD. Collectively, these studies reveal that the evolution of SSD has been accompanied by underlying changes in the effect of testosterone on male growth, potentially facilitating the rapid evolution of SSD. PMID- 19549144 TI - Density dependence and climate effects in Rocky Mountain elk: an application of regression with instrumental variables for population time series with sampling error. AB - 1. Sampling error in annual estimates of population size creates two widely recognized problems for the analysis of population growth. First, if sampling error is mistakenly treated as process error, one obtains inflated estimates of the variation in true population trajectories (Staples, Taper & Dennis 2004). Second, treating sampling error as process error is thought to overestimate the importance of density dependence in population growth (Viljugrein et al. 2005; Dennis et al. 2006). 2. In ecology, state-space models are used to account for sampling error when estimating the effects of density and other variables on population growth (Staples et al. 2004; Dennis et al. 2006). In econometrics, regression with instrumental variables is a well-established method that addresses the problem of correlation between regressors and the error term, but requires fewer assumptions than state-space models (Davidson & MacKinnon 1993; Cameron & Trivedi 2005). 3. We used instrumental variables to account for sampling error and fit a generalized linear model to 472 annual observations of population size for 35 Elk Management Units in Montana, from 1928 to 2004. We compared this model with state-space models fit with the likelihood function of Dennis et al. (2006). We discuss the general advantages and disadvantages of each method. Briefly, regression with instrumental variables is valid with fewer distributional assumptions, but state-space models are more efficient when their distributional assumptions are met. 4. Both methods found that population growth was negatively related to population density and winter snow accumulation. Summer rainfall and wolf (Canis lupus) presence had much weaker effects on elk (Cervus elaphus) dynamics [though limitation by wolves is strong in some elk populations with well-established wolf populations (Creel et al. 2007; Creel & Christianson 2008)]. 5. Coupled with predictions for Montana from global and regional climate models, our results predict a substantial reduction in the limiting effect of snow accumulation on Montana elk populations in the coming decades. If other limiting factors do not operate with greater force, population growth rates would increase substantially. PMID- 19549145 TI - Pulsed resources and climate-induced variation in the reproductive traits of wild boar under high hunting pressure. AB - 1. Identifying which factors influence age and size at maturity is crucial for a better understanding of the evolution of life-history strategies. In particular, populations intensively harvested, hunted or fished by humans often respond by displaying earlier age and decreased size at first reproduction. 2. Among ungulates wild boar (Sus scrofa scrofa L.) exhibit uncommon life-history traits, such as high fertility and early reproduction, which might increase the demographic impact of varying age at first reproduction. We analysed variation in female reproductive output from a 22-year long study of an intensively hunted population. We assessed how the breeding probability and the onset of oestrus responded to changes of female body mass at different ages under varying conditions of climate and food availability. 3. Wild boar females had to reach a threshold body mass (27-33 kg) before breeding for the first time. This threshold mass was relatively low (33-41% of adult body mass) compared to that reported in most other ungulates (about 80%). 4. Proportions of females breeding peaked when rainfall and temperature were low in spring and high in summer. Climatic conditions might act through the nutritional condition of females. The onset of oestrus varied a lot in relation to resources available at both current and previous years. Between none and up to 90% of females were in oestrus in November depending on the year. 5. Past and current resources accounted for equivalent amount of observed variations in proportions of females breeding. Thus, wild boar rank at an intermediate position along the capital-income continuum rather than close to the capital end where similar-sized ungulates rank. 6. Juvenile females made a major contribution to the yearly reproductive output. Comparisons among wild boar populations facing contrasted hunting pressures indicate that a high demographic contribution of juveniles is a likely consequence of a high hunting pressure rather than a species-specific life-history pattern characterizing wild boar. PMID- 19549146 TI - Variation of adult survival drives population dynamics in a migrating forest bat. AB - 1. Variation of survival across time, between sex and ages strongly affect the population dynamics of long-lived species. Bats are extremely long-lived, but the variation of their survival probabilities is poorly studied with reliable methods. 2. We studied annual local survival probabilities of the migratory Leisler's bats Nyctalus leisleri based on capture-recapture data from 1119 individuals sampled in bat boxes over 20 years in eastern Germany. We assessed variation in survival between sex and age classes, estimated the temporal variance of survival and tested whether survival was affected by weather during hibernation or pregnancy. 3. Among females, our analyses revealed two groups of individuals present with different roosting occupancy, survival and/or dispersal. Local survival of locally born females increased with age [first year: 0.45 +/- 0.04 (SE); later: 0.76 +/- 0.04] and the high recapture probabilities indicate regular presence in the roosts. Recapture probabilities and local survival of foreign adult females were significantly lower, indicating less frequent presence in the roosts and stronger dispersal from the study area. 4. In adult males, locally born and foreign individuals were nearly identical regarding survival and recapture, indicating a more homogenous group. Local survival was very low in the first year (0.04 +/- 0.08), most likely caused by strong natal dispersal. It further increased with age (second year: 0.55 +/- 0.20, later: 0.69 +/- 0.07). 5. Survival probabilities of all females varied significantly and in parallel across time, suggesting that a common environmental factor was operating which affected all individuals similarly. Spring temperature and winter North Atlantic Oscillation explained maximally 9% each of the variation in first year and adult female survival. In contrast to our expectations, the temporal variance of first year survival was lower than that of adult survival. 6. We found evidence of a complicated social population structure of female Leisler's bats. Our analyses suggest that their population dynamics are driven to a large amount by variation of survival, in particular by adult survival. The reason for the major temporal variations remains to be identified. PMID- 19549148 TI - Viability of human fibroblasts in coconut water as a storage medium. AB - AIM: To evaluate the effectiveness of a new storage medium for avulsed teeth, coconut water, in maintaining the viability of human fibroblasts. METHODOLOGY: Cell viability after different time periods was evaluated in the following storage media: coconut water, coconut water with sodium bicarbonate, milk, saline and still mineral water. Human fibroblasts were seeded in Eagle's minimal essential medium (EMEM) supplemented with 7.5% foetal calf serum. After trypsinisation, 100 microL of culture medium containing approximately 10(4) cells mL(-1) were collected and pipetted into the wells of 96-well plates, which were incubated overnight in 5% CO(2) and 95% air mixture at 37 degrees C. EMEM was then replaced by the storage media and the plates were incubated at 37 degrees C for 1, 2 and 4 h. Cell viability was determined using the neutral red assay. The proportions of viable cells after exposure to the storage media were analysed statistically by anova and the least significant difference (LSD) test (alpha = 5%). RESULTS: Milk had the greatest capacity to maintain cell viability (P < 0.05), followed by coconut water with sodium bicarbonate and saline. Coconut water was significantly worse at maintaining cell viability compared to milk, coconut water with sodium bicarbonate and saline. The smallest number of viable cells was observed for mineral water (P < 0.05). CONCLUSION: Coconut water was worse than milk in maintaining human fibroblast cell viability. PMID- 19549147 TI - Maternal investment in relation to sex ratio and offspring number in a small mammal - a case for Trivers and Willard theory? AB - 1. Optimal parental sex allocation depends on the balance between the costs of investing into sons vs. daughters and the benefits calculated as fitness returns. The outcome of this equation varies with the life history of the species, as well as the state of the individual and the quality of the environment. 2. We studied maternal allocation and subsequent fecundity costs of bank voles, Myodes glareolus, by manipulating both the postnatal sex ratio (all-male/all-female litters) and the quality of rearing environment (through manipulation of litter size by -2/+2 pups) of their offspring in a laboratory setting. 3. We found that mothers clearly biased their allocation to female rather than male offspring regardless of their own body condition. Male pups had a significantly lower growth rate than female pups, so that at weaning, males from enlarged litters were the smallest. Mothers produced more milk for female litters and also defended them more intensively than male offspring. 4. The results agree with the predictions based on the bank vole life history: there will be selection for greater investment in daughters rather than sons, as a larger size seems to be more influencial for female reproductive success in this species. Our finding could be a general rule in highly polygynous, but weakly dimorphic small mammals where females are territorial. 5. The results disagree with the narrow sense Trivers & Willard hypothesis, which states that in polygynous mammals that show higher variation in male than in female reproductive success, high-quality mothers are expected to invest more in sons than in daughters. PMID- 19549149 TI - Sealing ability, water sorption, solubility and toothbrushing abrasion resistance of temporary filling materials. AB - AIM: To evaluate marginal seal, water sorption, solubility and loss of mass after brushing of several temporary filling materials. METHODOLOGY: For marginal seal, Class I cavities, including endodontic access preparations, were made in human molar teeth and restored using one or other of several temporary filling materials (n = 10): zinc oxide/calcium sulphate-based cement (Cavit, 3M,ESPE, St. Paul, MN, USA), zinc oxide/eugenol cement (IRM, Dentsply Caulk, Milford, DE, USA), glass ionomer cement (Vidrion R, SSWhite, Rio de Janeiro, RJ, Brazil) or a dimethacrylate-based filling (Bioplic, Biodinamica, Londrina, PR, Brazil). Dye penetration was assessed after thermocycling and immersion in 0.5% basic fuchsine solution. For water sorption, solubility and loss of mass analyses, disc-shaped specimens were made. Water sorption and solubility were evaluated by mass alteration after storage in distilled water for 7 days (n = 7). Loss of mass was calculated based on the difference of mass after abrasion with a toothbrush (n = 5), and surfaces were analysed by SEM. Data of water sorption, solubility and loss of mass were submitted to anova and Tukey's test, and marginal sealing data to Kruskal-Wallis test (P < 0.05). RESULTS: Statistically significant differences were observed for marginal sealing (P < 0.0001), water sorption (P < 0.01), solubility (P < 0.01) and loss of mass (P < 0.05). Bioplic had the best marginal seal. Cavit had the greatest water sorption and solubility. Vidrion R and Bioplic had the lowest solubility. Loss of mass after brushing was higher for Cavit, followed by Bioplic, IRM and Vidrion R. Cavit and Vidrion R were worn aggressively by brushing. CONCLUSIONS: The resin-based temporary filling Bioplic produced the best marginal seal, and was associated with the lowest water sorption, solubility and loss of mass. PMID- 19549150 TI - Accuracy of root canal length determination with the impedance ratio method. AB - AIM: To provide insight into the principles of operation of electronic apex locators, determine optimal measuring parameters of the impedance ratio method, and to evaluate its accuracy. METHODOLOGY: Electrical impedance was measured ex vivo on 14 extracted human teeth using a QuadTech 1920 precision impedance analyzer. A file electrode was inserted into the root canal; the second electrode was placed in the saline solution surrounding the tooth. Measurements were performed in a frequency range from 20 Hz to 1 MHz, and repeated with different distances of the file tip from the major apical foramen. The measured impedances were analysed as a function of distance of the file tip to the major apical foramen. Parameters (e.g. measurement frequencies, impedance ratio value) that would result in optimal working length determination were evaluated. RESULTS: The optimal determination of the major apical foramen position was obtained at frequencies of 5 kHz and 0.5 kHz, 10 kHz and 0.5 kHz, and 5 kHz and 1 kHz, for the impedance ratios 0.73 (95% CI: -0.33 to 1.74 mm), 0.66 (95% CI: -0.34 to 1.81 mm) and 0.79 (95% CI: -0.33 to 1.58 mm) respectively. The limit of +/-0.5 mm was attained in 86% of all measurements. Standard deviations decreased as the average measured distance approached and extended beyond the major apical foramen. CONCLUSIONS: With the obtained optimal measuring parameters, the impedance ratio method determined position of the major foramen within +/-0.5 mm. Accuracy varied depending on the set of frequencies used for evaluation as well as on the selected impedance ratio. PMID- 19549151 TI - The nociceptive and anti-nociceptive effects of white mineral trioxide aggregate. AB - AIM: To assess the nociceptive and antinociceptive effects of white mineral trioxide aggregate (WMTA) using the orofacial formalin test in rats. METHODOLOGY: Rats (n = 10 in each group) were separately injected into the ipsilateral upper lip with either 40 microL of a 2.5% formalin solution and eugenol (50 mg kg(-1)) or WMTA (5, 10 and 20 mg dissolved in 0.2 mL saline) alone. In a second experiment to evaluate antinociception effects, 15 min prior to formalin injection, rats were pre-treated with either white ProRoot MTA (20 mg dissolved in 0.2 mL saline) or eugenol. The time each rat spent rubbing the injected site with its paw, as an index of nociception, was recorded for a period of 45 min. RESULTS: Administration of 40 microL white ProRoot MTA (5, 10 and 20 mg per 0.2 mL) alone did not produce any significant nociceptive response. Moreover, prior treatment with WMTA caused significant (P < 0.001) inhibition of formalin-induced nociception. Injection of eugenol (50 mg kg(-1)) provoked the first phase of a nociceptive response, although its intensity was reduced compared with that produced by formalin. Pre-treatment with eugenol significantly (P < 0.0001) inhibited the induction of nociception by formalin. Comparison of the behavioural responses observed in WMTA and eugenol-treated rats alone or in combination with formalin revealed that WMTA did not only induce pain behaviour but also prevented formalin-induced nociception. CONCLUSION: White mineral trioxide aggregate, when compared with eugenol, was more effective in treating nociceptive pain in the orofacial formalin test. PMID- 19549152 TI - A radiographic assessment of the prevalence of pulp stones in a group of Turkish dental patients. AB - AIM: To determine the prevalence of pulp stones in a group of patients using radiographs, and to assess any associations with age, gender, systemic diseases, tooth type, jaw, caries, restorations, impacted third molars and dental anomalies including dens invaginatus, taurodontism, dens evaginatus and microdontia. METHODOLOGY: A total of 519 patients were selected randomly amongst patients referred to Oral Diagnosis and Radiology Department, Faculty of Dentistry, Ankara University. After clinical inspection, full-mouth periapical radiographs were obtained. An oral radiologist examined the radiographs to identify pulp stones. Frequency distribution of teeth with pulp stones was calculated and statistical analysis was performed with chi-square test. RESULTS: Of the patients, 313 (60%) were female and 206 (40%) were male. Sixty (12%) had one or more teeth that contained pulp stones. Amongst the 13,474 teeth examined radiographically, 627 (5%) had pulp stones. There was no significant association between pulp stone occurrence and gender or systemic diseases. As age increased, the prevalence of pulp stones increased (P < 0.01). Molars had statistically more pulp stones than premolars and incisors (P < 0.001). Frequencies in both maxillary and mandibular jaw were similar. There was no association between pulp stones and impacted third molars, caries, restorations or dental anomalies. CONCLUSIONS: The prevalence of pulp stones increased with age and was significantly higher in molar teeth than premolars and incisors. There was no significant association between pulp stones and gender, systemic diseases, impacted third molars, condition of the crown and dental anomalies. PMID- 19549153 TI - Root and canal morphology of mandibular first and second molar teeth in a Jordanian population. AB - AIM: To examine the root and canal morphology of mandibular permanent first and second molar teeth in a Jordanian population. METHODOLOGY: A total of 685 extracted mandibular first and second permanent molar teeth were collected from dental clinics within north Jordan. The teeth were examined visually and the root number and morphology were recorded. After that, access cavities were prepared, pulp tissue dissolved by sodium hypochlorite and the root canals injected with Indian ink. Stained teeth were decalcified with 10% nitric acid, dehydrated with ascending concentrations of alcohol and rendered clear by immersion in methyl salicylate. The following observations were evaluated: (i) number of canals per root; (ii) number of root canals per tooth; (iii) canal configuration in each root; (iv) number and location of lateral canals and (v) presence of intercanal communications. RESULTS: Of 330 mandibular first molars, the majority had three (48%) or four (46%) canals, whilst 4% had a third disto-lingual root. Of 355 mandibular second molars, 58% had three canals, 19% two and 17% had four canals, whilst 10% had C-shaped roots. The most prevalent canal configuration in the mesial root of both first (53%) and second (40%) molars was type IV, and in distal roots was type I (54% in first and 79% in second molars). CONCLUSIONS: Jordanian mandibular first and second molar teeth exhibit features close to the average Caucasian root and canal morphology. PMID- 19549154 TI - Apexification: the beginning of its end. AB - Apexification is a procedure for treating and preserving immature permanent teeth that have lost pulp vitality. It contrasts apexogenesis in terms of its outcome in that apical maturation and normal root thickness cannot be obtained. Apexification has been a routine practice for such teeth for many decades, and despite a literature replete with discussion, including recent artificial barrier methods with mineral trioxide aggregate, ultimately there has been no major breakthrough to improve this treatment. Recently, two new clinical concepts have emerged. One involves a revitalization approach to achieve tissue generation and regeneration. In this method, new living tissue is expected to form in the cleaned canal space, allowing continued root development in terms of both length and thickness. The other is the active pursuit of pulp/dentine regeneration via tissue engineering technology to implant or re-grow pulps. Although the technology is still at its infancy, it has the potential to benefit immature pulpless teeth by allowing continued growth and maturation. With this understanding, it may be predicted that apexification will become less needed in years to come. This study will overview the recent concept of pulp revitalization in the treatment of immature teeth with nonvital pulps and the emerging research on pulp tissue engineering and regeneration. PMID- 19549155 TI - Effect of damping properties on fracture resistance of root filled premolar teeth: a dynamic finite element analysis. AB - AIM: To evaluate the ex vivo effects of damping on stress concentration in root filled premolar teeth. METHODOLOGY: Damping ratios of maxillary premolar teeth that had undergone root canal treatment were tested in a laboratory model. In addition, two-dimensional finite element (FE) models were established for dynamic analysis. RESULTS: The mean-damping ratio was significantly lower in premolar teeth that had undergone root canal preparation (8.50 +/- 0.53%) than in unprepared teeth (14.42 +/- 2.17%) (P < 0.05). However, root filling had a significant positive effect on the damping ratio of the tooth (10.84 +/- 1.70%) (P < 0.05). When the damping ratio was taken into consideration, FE analysis revealed that peak stresses in the apical one-third of the root on the buccal side were reduced by 31.8% when mastication forces were applied on the palatal cusp and occlusal fossa. CONCLUSION: Pulp tissue plays an important role in providing protective effects when teeth are subjected to a dynamic load. However, root filled teeth do not provide such protective effects. PMID- 19549156 TI - Headache prophylaxis with BoNTA: patient characteristics. AB - OBJECTIVE: To assess the characteristics of patients receiving botulinum toxin type A (BoNTA; BOTOX((R))) in the treatment of headache (HA) disorders. METHODS: The following observational epidemiologic data and baseline patient characteristics were prospectively collected from eligible patients treated with BoNTA at 10 US HA specialty centers: demographics; HA diagnoses and characteristics (frequency, severity, and disability); prior and current HA treatments and response; clinical response to BoNTA; Migraine Disability Assessment (MIDAS) questionnaire; and adverse events. Patients maintained a daily HA diary and were evaluated at each follow-up visit. RESULTS: Of 703 patients enrolled (mean age 43.1 years, 78.5% females, 95.4% white), nearly 66% had a diagnosis of chronic migraine (CM), with or without medication overuse. Approximately 75% had severe disability (MIDAS grade IV), and the mean pain rating was 6.5 (where 0 = no pain, 10 = pain as bad as it can be). More than 90% of patients had >or=1 prophylactic HA treatment failure; median number of failures was 4. Significant association was observed between HA frequency and MIDAS grade (P < .001). Approximately 80% of patients with CM had severe (grade IV) disability. The median number of monthly medication days was higher in the group with MIDAS grade IV (P < .001). HA frequency and severity, failed prophylactic therapies, and greater number of coexisting medical conditions were all negatively associated with measures of health-related quality of life. CONCLUSIONS: Majority of patients treated with BoNTA in a specialty HA center presented with a CM diagnosis. HA disability was correlated with measures of frequency and treatment utilization. PMID- 19549157 TI - A comment on remarks regarding Schoonman et al. "Migraine headache is not associated with cerebral or meningeal vasodilatation--a 3T magnetic resonance angiography study". PMID- 19549158 TI - Recurrent coital headache associated with an unruptured carotid saccular aneurysm. AB - We report the case of a 48-year-old woman whose recurrent coital headache ceased following intracranial internal carotid artery aneurysm embolization. PMID- 19549159 TI - Stress management for migraine: recent research and commentary. PMID- 19549161 TI - Recent studies on stress management-related treatments for migraine. PMID- 19549160 TI - Epidemiology and neurobiology of stress and migraine. PMID- 19549162 TI - Rapid oral transmucosal absorption of sumatriptan, and pharmacodynamics in acute migraine. AB - OBJECTIVES: (1) To determine whether sumatriptan can be absorbed across the oral mucosa, and, if so; then (2) to describe its pharmacokinetics; and (3) to investigate whether there are pharmacodynamic correlates of such pharmacokinetics in patients experiencing migraine attacks. METHODS: Two clinical trials. The first, in normal volunteers, compared the pharmacokinetic performance of a lingual spray (LS) formulation of sumatriptan (2 dose sizes, one of which in both the fed and fasted state) with an orthodox 50-mg sumatriptan tablet. The second clinical trial, in a patient population enriched by documenting suboptimal response to an initial 50-mg sumatriptan tablet, was a multiple-attack, crossover, fixed dose-order, open-label comparison of sumatriptan administered by LS (up to 3 different dose sizes) and a 100-mg sumatriptan tablet. RESULTS: The LS formulations resulted in double-peaked time-plasma concentration curves that are consistent with absorption of sumatriptan across the oral mucosa. The first T(max) was usually about 10-15 minutes. In the enriched patient population, this corresponded with evidence of earlier efficacy for the LS in comparison with a 50 mg tablet; the lower dose size for the former was consistent with oral mucosal drug absorption, and evasion of first-pass metabolism. CONCLUSIONS: The initial pharmacokinetics of LS approximate to those of a subcutaneous injection, albeit some fraction of these doses is also swallowed. These pharmacokinetics correspond with earlier effectiveness of LS in comparison with a 50-mg sumatriptan tablet, and at lower dose, in an enriched, relevant patient population. These initial studies support further development of this innovative formulation of sumatriptan and this new route of administration. PMID- 19549163 TI - Migrainous vertigo responsive to intravenous methylprednisolone: case reports. AB - Migrainous vertigo (MV) is a newer evolving concept in which vertigo is causally related to migraine. We report 4 patients with MV. Episodes of severe MV of more than 1-day duration were successfully terminated by intravenous methylprednisolone (IV MPS) in 2 patients. Two other patients who had attacks of MV almost daily also showed complete response to IV MPS. PMID- 19549164 TI - Primary hyperoxaluria causing ESRD and gangrene of extremities leading to amputation. AB - Primary hyperoxaluria is an uncommon, inherited metabolic disorder due to hepatic enzyme deficiencies with consequent hepatic oxalate overproduction and attendant systemic complications. The diagnosis is established on a combination of clinical parameters, elevated urinary excretion of oxalate and glycolate and determination of alanine glyoxylate aminotransferase in the liver tissue. We describe a 45-year old female with end-stage renal disease secondary to nephrolithiasis, who presented with a fulminating vascular syndrome before confirming the diagnosis of primary hyperoxaluria. This case illustrates that in this infrequent clinical entity, the diagnosis is often delayed with incorrect initial management. PMID- 19549165 TI - Small insertion (c.869insC) within F13A gene is dominant in Tunisian patients with inherited FXIII deficiency due to ancient founder effect. PMID- 19549166 TI - Antiphospholipid antibodies in haemophilia patients with severe bleeding tendency: cause, consequence or a consequential cause? AB - The prevalence, cause and the impact of antiphospholipid antibodies (APAs) on the clinical severity in haemophilia patients is poorly studied. We studied 72 severe seronegative (negative for HIV, HBsAg, HCV) haemophilia patients for the presence of four common APAs. Twenty-six (36.1%) were positive for any one of the APAs studied of which eight were positive only for anticardiolipin antibodies, three for beta2 glycoprotein (beta2GP1), four for prothrombin (PT) and six for anti annexin antibodies. Remaining six patients showed multi-specific antibodies. Further, clinically severe haemophilia patients (n = 37) showed higher prevalence of APAs as compared with the clinically milder group (n = 35) suggesting that these antibodies do not contribute in alleviating the clinical severity in haemophilia patients as has been observed with other inherited thrombophilia markers. The study of in vitro thrombin generation showed a higher endogenous thrombin potential (ETP) i.e. almost normal, in case of beta2GP1-positive patients as compared with patients with other types of APAs. High prevalence of APAs in clinically severe haemophilia patients may be a consequence of continuing tissue damage in the clinically severe group; as in India, clotting factor concentrates cannot be used ad lib because of financial constraints. Higher thrombin-generating potential in case of patients positive for beta2GP1 did not seem to have any impact on the clinical severity of haemophilia patients. PMID- 19549167 TI - Versatility of the capsular genes during biofilm formation by Streptococcus pneumoniae. AB - Streptococcus pneumoniae forms part of the natural microbiota of the nasopharynx. For the pneumococcus to cause infection, colonization needs to occur and this process is mediated by adherence of bacteria to the respiratory epithelium. Although the capsular polysaccharide (CPS) of S. pneumoniae is known to be important for infection to occur, its role in colonization is controversial. Biofilm models are starting to emerge as a promising tool to investigate the role of CPS during nasopharyngeal carriage, which is the first step in the dissemination and initiation of a pneumococcal infection. Using a well-defined model system to analyse in vitro biofilm formation in pneumococcus, here we explore the molecular changes underlying the appearance of capsular mutants using type 3 S. pneumoniae cells. Spontaneous colony phase variants show promoter mutations, as well as duplications, deletions and point mutations in the cap3A gene, which codes for a UDP-glucose dehydrogenase (UDP-GlcDH). Increased biofilm forming capacity could usually be correlated with a reduction both in colony size and in the relative amount of CPS present on the cell surface of each colony variant. However, a mutation in Cap3A Thr83Ile (a strictly conserved residue in bacterial UDP-GlcDHs) that resulted in very low CPS production also led to impaired biofilm formation. We propose that non-encapsulated mutants of pneumococcal type 3 strains are essentially involved in the initial stages (the attachment stage) of biofilm formation during colonization/pathogenesis. PMID- 19549169 TI - Lag-phase autophagy in the methylotrophic yeast Pichia pastoris. AB - When microbes sense environmental changes, they often temporarily attenuate cell growth to adapt to the new situations, showing a lag phase. In this study, we report that the methylotrophic yeast, Pichia pastoris, induced autophagy during the lag phase after the cells were shifted from glucose to methanol medium. Through the autophagic process at least two proteins, aminopeptidase I precursor and cytosolic aldehyde dehydrogenase, were found to be transported into the vacuole, which was dependent on PpAtg11 and PpAtg17, respectively. Notably, PpAtg1 and PpAtg17 were required for early exit from the lag phase during the methanol adaptation. In accordance, phosphorylation states of elongation initiation factor 2alpha indicated reductions of intracellular amino-acid pools in the atg mutant strains. Together, these data demonstrate the importance of amino acid recycling by autophagy during a cell-remodeling process. PMID- 19549168 TI - Dissection of sigma(E)-dependent cell lysis in Escherichia coli: roles of RpoE regulators RseA, RseB and periplasmic folding catalyst PpiD. AB - To understand the mechanism of sigma(E)-dependent cell lysis, we examined the consequences of deletion derivatives of rpoE regulators rseA, rseB and rseC on sigma(E) transcription, on levels of free versus membrane-bound sigma(E) and on OMP-biogenesis limiting factor(s) that could impact cell lysis. RT-PCR showed that individual nonpolar DeltarseA and DeltarseB increased the rpoE expression to varying extents, with pronounced induction in DeltarseA. Significantly the ratio of soluble (free) versus membrane-bound form of RpoE increased in DeltarseA, however without increase of its total amount, unraveling furthermore complexity in RpoE regulation. Significant characteristics of cell lysis, accompanied by a severe reduction in the levels of periplasmic OMP-folding factor (PpiD), were observed in DeltarseA. The cell-lysis phenotype of DeltarseA was suppressed by either rseA or ppiD plasmids, but neither by rseB nor by rseC clones. However, the cell lysis of the wild-type strain was almost completely repressed not only by the rseA clone but also by the rseB clone, suggesting RseB might be limiting in vivo. Thus, increase in the ratio of free sigma(E) in rseA mutants with a concomitant reduction in PpiD levels can account for sigma(E)-dependent lysis in concert with a potential role of small RNAs on the lysis process. PMID- 19549170 TI - Intracellular polarity protein PAR-1 regulates extracellular laminin assembly by regulating the dystroglycan complex. AB - Cell polarity depends on extrinsic spatial cues and intrinsic polarity proteins including PAR-aPKC proteins. In mammalian epithelial cells, cell-cell contacts provide spatial cues that activate the aPKC-PAR-3-PAR-6 complex to establish the landmark of the initial cellular asymmetry. PAR-1, a downstream target of the aPKC-PAR-3-PAR-6 complex, mediates further development of the apical and basolateral membrane domains. However, the relationships between the PAR-aPKC proteins and other extrinsic spatial cues provided by the extracellular matrix (ECM) remain unclear. Here, we show that PAR-1 colocalizes with laminin receptors and is required for the assembly of extracellular laminin on the basal surface of epithelial cells. Furthermore, PAR-1 regulates the basolateral localization of the dystroglycan (DG) complex, one of the laminin receptors essential for basement membrane formation. We also show that PAR-1 interacts with the DG complex and is required for the formation of a functional DG complex. These results reveal the presence of a novel inside-out pathway in which an intracellular polarity protein regulates the ECM organization required for epithelial cell polarity and tissue morphogenesis. PMID- 19549171 TI - ATP-induced osteoclast function: the formation of sealing-zone like structure and the secretion of lytic granules via microtubule-deacetylation under the control of Syk. AB - Osteoclasts are bone-resorbing cells which play an exclusive role in bone remodeling, but the molecular mechanisms of osteolysis, how osteoclasts are activated and how the lytic granules are finally released towards the bone matrix are poorly understood. Here we show that an energy molecule ATP induces osteolysis via P2X(7)-nucleotide receptor and that deacetylation of alpha-tubulin is essential for the whole process of osteolysis under the control of a tyrosine kinase Syk. By developing a traceable and reproducible in vitro analyzing system for osteoclast function, we found that ATP-signaling gives rise to two events simultaneously (i) cytoskeletal reorganization for the formation of sealing zones, ring-like adhesion structures which delimit the contact surface, and (ii) the delivery and secretion of lytic granules towards the delimited site on the matrix. We further found that deacetylation of alpha-tubulin is a critical reaction for osteoclast function. Pharmacological inhibition of alpha-tubulin deacetylation resulted in (i) failure of the sealing-zone like structure formation and (ii) ceased secretion of lytic granules. Additionally, kinetics of deacetylation was found to be regulated by Syk. These data suggest a novel P2X(7) microtubular regulation pathway related to Syk for a therapeutic target in osteolytic diseases. PMID- 19549172 TI - Comparative in vivo and in vitro analyses of putative virulence factors of Burkholderia pseudomallei using lipopolysaccharide, capsule and flagellin mutants. AB - Burkholderia pseudomallei is a gram-negative bacillus that is the causative agent of melioidosis. We evaluated host-pathogen interaction at different levels using three separate B. pseudomallei mutants generated by insertional inactivation. One of these mutants is defective in the production of the polysaccharide side chains associated with lipopolysaccharide; one does not produce the capsular polysaccharide with the structure -3)-2-O-acetyl-6-deoxy-beta-d-manno heptopyranose-(1-; and the third mutant does not produce flagellin. We compared the in vivo virulence in BALB/c mice, the in vitro fate of intracellular survival inside human polymorphonuclear cells (PMNs) and macrophages (Mphis) and the susceptibility to killing by 30% normal human serum, reactive nitrogen and oxygen intermediates and antimicrobial peptides with that of their wild-type counterpart. The lipopolysaccharide and capsule mutants demonstrated a marked reduction in virulence for BALB/c mice, but the flagellin mutant was only slightly less virulent than the parent strain. The results from the BALB/c mice experiments correlated with survival in Mphis. The lipopolysaccharide and capsule mutants were also more susceptible to killing by antimicrobial agents. All bacteria were equally susceptible to killing by PMNs. Altogether, the data suggest that lipopolysaccharide and capsule and, to a much lesser extent, flagella, are most likely associated with the virulence of this bacterium and highlight the importance of intracellular killing by PMNs and Mphis in disease pathogenesis. PMID- 19549173 TI - Suppression of inflammatory responses by celastrol, a quinone methide triterpenoid isolated from Celastrus regelii. AB - BACKGROUND: Celastrol, a quinone methide triterpenoid isolated from the Celastraceae family, exhibits various biological properties, including chemopreventive, antioxidant and neuroprotective effects. In this study, we showed that celastrol inhibits inflammatory reactions in macrophages and protects mice from skin inflammation. MATERIALS AND METHODS: Anti-inflammatory effects of celastrol (0-1 microM) were examined in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. To investigate the effects of celastrol (0-50 microg per mice) in vivo, activation of myeloperoxidase (MPO) and histological assessment were examined in the 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear oedema model. RESULTS: Our in vitro experiments showed that celastrol suppressed not only LPS-stimulated generation of nitric oxide and prostaglandin E(2), but also expression of inducible nitric oxide synthase and cyclooxygenase-2 in RAW264.7 cells. Similarly, celastrol inhibited LPS-induced production of inflammatory cytokines, including tumour necrosis factor-alpha and interleukin-6. In an animal model, celastrol protected mice from TPA-induced ear oedema, possibly by inhibiting MPO activity and production of inflammatory cytokines. CONCLUSIONS: Our data suggest that celastrol inhibits the production of inflammatory mediators and is a potential target for the treatment of various inflammatory diseases. PMID- 19549174 TI - Patient-orientated web sites on laryngectomy: is their information readable? AB - The objective of the study was to determine levels of readability of commonly accessed websites containing patient information on laryngectomy. A Cross sectional study of Internet websites was designed. The first 20 websites obtained from a Google search of the word "laryngectomy" that contained a patient information section were selected. Primary outcome measures were the Flesch Readability Ease Score (FRE) and Flesch-Kincaid readability grade (FKRG) score; from these data UK reading age was calculated. The secondary outcome measure was whether or not a site was accredited by an online readability organisation. The reading ages of the 20 sites ranged from 7.8 to 14.7 years with a median of 11.7 years. Half of the Flesch Reading Ease scores were in the "Difficult" or "Fairly difficult" category with 30% falling into the "Standard" or "Fairly easy" categories. Only 20% sites fell into the "Easy" or "Very Easy" categories that are the recommended level for comprehension by the general UK population. Sites not accredited by an online healthcare quality and content control organisation had worse readability scores than those that were not (FRE: p = 0.007, FKRG: p = 0.012). The poor readability of many of the encountered sites about laryngectomy on the Internet may confuse patients who turn to the Internet for information. Methods to improve patient information websites are discussed. PMID- 19549175 TI - Successful treatment of depressed scars of the forehead secondary to herpes zoster using subdermal minimal surgery technology. PMID- 19549176 TI - Uncompleted fan flap for full-thickness lower lip defect. PMID- 19549177 TI - Nonablative fractional photothermolysis for the treatment of striae rubra. PMID- 19549178 TI - Combination treatment with basic fibroblast growth factor and artificial dermis improves complex wounds in patients with a history of long-term systemic corticosteroid use. PMID- 19549179 TI - Development of cerebral air emboli during mohs micrographic surgery. PMID- 19549180 TI - Adenoid Basal cell carcinoma simulating ameloblastoma. PMID- 19549181 TI - Complete closure using a double purse-string closure for skin defects. AB - BACKGROUND: The purse-string closure is useful to repair the round or oval defect. However, difficulty may exist in the achievement of complete closure of the relatively large defect. OBJECTIVE: A new technique was proposed which could redistribute the tension around the defect and achieve a complete and simple closure. METHODS AND MATERIALS: For 17 surgical defects following skin tumor, the double purse-string technique was performed. A midline interrupt suture was placed vertical to the skin relax tension line. Then, the original defect was split into halves and both smaller defects were closed with the pure-string suture. RESULTS: All skin defects were closed completely, and there were no complication occurrences for the repair postoperatively. The final cosmetic appearance was excellent. CONCLUSION: The double purse-string suture is a useful technique to achieve complete closure of the defect with a resulting excellent appearance. The authors have indicated no significant interest with commercial supporters. PMID- 19549182 TI - Novel pretrichial browlift technique and review of methods and complications. AB - BACKGROUND: The upper third of the face is integral to our perception of youth and beauty. While the eyelids anchor this facial cosmetic unit, the eyebrows and forehead are intrinsically linked to the upper eyelids, and their position and texture play an important role in creating pleasing eyes as well as conveying mood and youth. The most common browlifts are performed with endoscopic visualization. Yet, this technique requires special equipment and a prolonged learning curve. OBJECTIVE: To demonstrate a novel pretrichial technique and to review different browlift methods and their potential adverse effects. METHODS Case series and review of the literature. RESULTS: The pretrichial browlift results in a mild to moderate browlift with secondary smoothing of the forehead topography. Aside from bruising and swelling, it results in minimal adverse effects. Other techniques are also effective but may create a larger scar such as a direct browlift, may be more difficult in terms of approach such as the browpexy, or require endoscopes. CONCLUSION: Browlifts are an important procedure in rejuvenating the upper third of the face and improving the overall facial aesthetic appearance. The pretrichial browlift is a less invasive open technique that is safe and effective for the appropriate patient. The authors have indicated no significant interests with commercial supporters. PMID- 19549183 TI - Method for facilitating the application of digital tourniquets. AB - The authors have indicated no significant interests with commercial supporters. PMID- 19549184 TI - Bilobed flap for full-thickness nasal defect: a common flap for an uncommon indication. PMID- 19549185 TI - Treatment of punched-out atrophic and rolling acne scars in skin phototypes III, IV, and V with variable square pulse erbium:yttrium-aluminum-garnet laser resurfacing. AB - BACKGROUND: Treatment of acne scars remains a challenge, especially in dark skinned individuals. Treatment parameters may be optimized by selecting appropriate pulse width and laser energy that enhance tissue thermal response with limited morbidity. OBJECTIVE: To determine the efficacy and side effects of variable square pulse (VSP) erbium:yttrium-aluminum-garnet (Er:YAG) laser resurfacing for treatment of punched-out atrophic and rolling acne scars. METHODS Twenty-four subjects with acne scars were treated monthly for 2 months with four passes of VSP Er:YAG laser resurfacing using a 7-mm spot size and a fluence of 0.4 J/cm(2). Subjects were divided into two groups and treated with two different pulse widths: 300 micros (short pulse, SP) and 1,500 micros (extra-long pulse, XLP). Objective and subjective assessments were obtained at baseline and 1, 2, and 4 months after treatment. RESULTS: In the SP group, skin smoothness improved significantly (p<.01); in the XLP group, skin smoothness (p<.05) and scar volume (p<.05) improved significantly from baseline. Adverse effects consisted of transient postinflammatory hyperpigmentation (18%) and acneiform eruption (9%). CONCLUSIONS: Low-fluence VSP Er:YAG laser resurfacing is a promising treatment option for acne scars, with minimal risk of side effects. Laser pulse width and energy determine the efficacy and the risk of side effects. The authors have indicated no significant interests with commercial supporters. PMID- 19549186 TI - Randomized, placebo-controlled study of a new botulinum toxin type a for treatment of glabellar lines: efficacy and safety. AB - BACKGROUND: A new botulinum toxin type A (BoNT-A) has been assessed in the United States for treatment of glabellar lines. In April 2009, the US FDA approved the Biologics License Application for a new US formulation of BoNT-A (Dysport [abobotulinum toxin A]; Medicis Aesthetics Inc., Scottsdale, AZ). OBJECTIVE: To compare efficacy and safety of a single treatment of BoNT-A with placebo in subjects with moderate to severe glabellar lines. METHODS AND MATERIALS: One hundred fifty-eight subjects with moderate to severe glabellar lines were randomized 2:1 to receive 50 U of BoNT-A (n=105) or placebo (n=53). Responders were defined as having no or mild glabellar lines at 30 days posttreatment according to investigator and subject assessments (co-primary endpoint) using the validated Glabellar Line Scale Score at maximum frown. Subject diaries were used to document onset of effect. When conducting the research, the authors conformed to the ethical guidelines of the 1975 Declaration of Helsinki. RESULTS: According to investigator assessment, the proportion of responders to BoNT-A at Day 30 was 89.5%, versus 7.5% for placebo (p<.001); according to subject assessment, the proportion of responders was 75.7%, versus 9.8% for placebo (p<.001). CONCLUSION: A single treatment with BoNT-A (50 U) was significantly superior to placebo in the correction of moderate to severe glabellar lines, with comparable tolerability. PMID- 19549187 TI - Quenched hydrogen/deuterium exchange NMR characterization of amyloid-beta peptide aggregates formed in the presence of Cu2+ or Zn2+. AB - Alzheimer's disease, a neurodegenerative disorder causing synaptic impairment and neuronal cell death, is strongly correlated with aggregation of the amyloid-beta peptide (Abeta). Divalent metal ions such as Cu(2+) and Zn(2+) are known to significantly affect the rate of aggregation and morphology of Abeta assemblies in vitro and are also found at elevated levels within cerebral plaques in vivo. The present investigation characterized the architecture of the aggregated forms of Abeta(1-40) and Abeta(1-42) in the presence or absence of either Cu(2+) or Zn(2+) using quenched hydrogen/deuterium exchange combined with solution NMR spectroscopy. The NMR analyses provide a quantitative and residue-specific structural characterization of metal-induced Abeta aggregates, showing that both the peptide sequence and the type of metal ion exert an impact on the final architecture. Common features among the metal-complexed peptide aggregates are two solvent-protected regions with an intervening minimum centered at Asn27, and a solvent-accessible N-terminal region, Asp1-Lys16. Our results suggest that Abeta in complex with either Cu(2+) or Zn(2+) can attain an aggregation-prone beta-strand-turn-beta-strand motif, similar to the motif found in fibrils, but where the metal binding to the N-terminal region guides the peptide into an assembly distinctly different from the fibril form. PMID- 19549188 TI - The novel molecule 2-[5-(2-chloroethyl)-2-acetoxy-benzyl]-4-(2-chloroethyl) phenyl acetate inhibits phosphoinositide 3-kinase/Akt/mammalian target of rapamycin signalling through JNK activation in cancer cells. AB - Screening a compound library of compound 48/80 analogues, we identified 2-[5-(2 chloroethyl)-2-acetoxy-benzyl]-4-(2-chloroethyl)-phenyl acetate (E1) as a novel inhibitor of the phosphoinositide 3-kinase/Akt pathway. In order to determine the mechanism of action of E1, we analysed the effect of E1 on components of the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin (mTOR) pathway. E1 demonstrated dose-dependent and time-dependent repression of Akt and mTOR activity in prostate and breast cancer cell lines, PC-3 and MCF-7, respectively. Inhibition of Akt and mTOR activity by E1 also coincided with increased c-Jun NH2 terminal kinase (JNK) phosphorylation. However, the mode of action of E1 is different from that of the mTOR inhibitor rapamycin. Proliferation and cell cycle analysis revealed that E1 induced cell cycle arrest and cell death in PC-3 and MCF-7 cells. Moreover, pretreatment of cancer cells with the JNK inhibitor SP600125 abolished the repression of Akt and mTOR activity by E1, indicating that the inhibition of Akt and mTOR by E1 is mediated through JNK activation. Consistently, E1 repressed Akt and mTOR activity in wild-type and p38-null mouse embryonic fibroblasts (MEFs), but not in MEFs lacking JNK1/2, and JNK-null MEFs were less sensitive to the antiproliferative effects of E1. We further showed that E1 can function cooperatively with suboptimal concentrations of paclitaxel to induce cell death in PC-3 and MCF-7 cells. Taken together, these data suggest that E1 induces cancer cell death through the JNK-dependent repression of Akt and mTOR activity and may provide a valuable compound for further development and research. PMID- 19549189 TI - Serine hydroxymethyltransferase from Plasmodium vivax is different in substrate specificity from its homologues. AB - The putative gene of Plasmodium vivax serine hydroxymethyltransferase (PvSHMT; EC 2.1.2.1) was cloned and expressed in Escherichia coli. The purified enzyme was shown to be a dimeric protein with a monomeric molecular mass of 49 kDa. PvSHMT has a maximum absorption peak at 422 nm with a molar absorption coefficient of 6370 M(-1) x cm(-1). The K(d) for binding of the enzyme and pyridoxal-5-phosphate was 0.14 +/- 0.01 microM. An alternative assay for measuring the tetrahydrofolate dependent SHMT activity based on the coupled reaction with 5,10 methylenetetrahydrofolate reductase (EC 1.5.1.20) from E. coli was developed. PvSHMT uses a ternary-complex mechanism with a k(cat) value of 0.98 +/- 0.06 s( 1) and K(m) values of 0.18 +/- 0.03 and 0.14 +/- 0.02 mM for L-serine and tetrahydrofolate, respectively. The optimum pH of the SHMT reaction was 8.0 and an Arrhenius's plot showed a transition temperature of 19 degrees C. Besides L serine, PvSHMT forms an external aldimine complex with D-serine, L-alanine, L threonine and glycine. PvSHMT also catalyzes the tetrahydrofolate-independent retro-aldol cleavage of 3-hydroxy amino acids. Although L-serine is a physiological substrate for SHMT in the tetrahydrofolate-dependent reaction, PvSHMT can also use D-serine. In the absence of tetrahydrofolate at high pH, PvSHMT forms an enzyme-quinonoid complex with D-serine, but not with L-serine, whereas SHMT from rabbit liver was reported to form an enzyme-quinonoid complex with L-serine. The substrate specificity difference between PvSHMT and the mammalian enzyme indicates the dissimilarity between their active sites, which could be exploited for the development of specific inhibitors against PvSHMT. PMID- 19549191 TI - Head-torso-hand coordination in children with and without developmental coordination disorder. AB - AIM: This study investigated the nature of coordination and control problems in children with developmental coordination disorder (DCD). METHOD: Seven adults (two males, five females, age range 20-28 y; mean 23 y, SD 2 y 8 mo) and eight children with DCD (six males, two females, age range 7-9 y; mean 8 y, SD 8 mo), and 10 without DCD (seven males, three females, age range 7-9 y; mean 8 y, SD 7 mo) sat in a swivel chair and looked at or pointed to targets. Optoelectronic apparatus recorded head, torso, and hand movements, and the spatial and temporal characteristics of the movements were computed. RESULTS: Head movement times were longer (p<0.05) in children with DCD than in the comparison group, even in the looking task, suggesting that these children experience problems at the lowest level of coordination (the coupling of synergistic muscle groups within a single degree of freedom). Increasing the task demands with the pointing condition affected the performance of children with DCD to a much greater extent than the other groups, most noticeably in key feedforward kinematic landmarks. Temporal coordination data indicated that all three groups attempted to produce similar movement patterns to each other, but that the children with DCD were much less successful than age-matched children in the comparison group. INTERPRETATION: Children with DCD have difficulty coordinating and controlling single degree-of freedom movements; this problem makes more complex tasks disproportionately difficult for them. Quantitative analysis of kinematics provides key insights into the nature of the problems faced by children with DCD. PMID- 19549192 TI - Sex ratio of siblings of children diagnosed with autism spectrum disorder. PMID- 19549190 TI - Characterization of the angiogenic activity of zebrafish ribonucleases. AB - Ribonucleases identified from zebrafish possess angiogenic and bactericidal activities. Zebrafish RNases have three intramolecular disulfide bonds, a characteristic structural feature of angiogenin, different from the typical four disulfide bonds of the other members of the RNase A superfamily. They also have a higher degree of sequence homology to angiogenin than to RNase A. It has been proposed that all RNases evolved from these angiogenin-like progenitors. In the present study, we characterize, in detail, the function of zebrafish RNases in various steps in the process of angiogenesis. We report that zebrafish RNase-1, 2 and -3 bind to the cell surface specifically and are able to compete with human angiogenin. Similar to human angiogenin, all three zebrafish RNases are able to induce phosphorylation of extracellular signal-regulated kinase 1/2 mitogen activated protein kinase. They also undergo nuclear translocation, accumulate in the nucleolus and stimulate rRNA transcription. However, zebrafish RNase-3 is defective in cleaving rRNA precursor, even though it has been reported to have an open active site and has higher enzymatic activity toward more classic RNase substrates such as yeast tRNA and synthetic oligonucleotides. Taken together with the findings that zebrafish RNase-3 is less angiogenic than zebrafish RNase-1 and -2 as well as human angiogenin, these results suggest that zebrafish RNase-1 is the ortholog of human angiogenin and that the ribonucleolytic activity of zebrafish RNases toward the rRNA precursor substrate is functionally important for their angiogenic activity. PMID- 19549193 TI - Measuring the quality of life of children with cerebral palsy: comparing the conceptual differences and psychometric properties of three instruments. AB - AIM: To compare the conceptual differences, internal consistency, and validity of the Cerebral Palsy Quality of Life Questionnaire for Children (CP QOL-Child), the Child Health Questionnaire (CHQ), and a European generic health-related quality of life (HRQOL) questionnaire (10-domain version; KIDSCREEN-10) for children with cerebral palsy (CP). METHOD: Two hundred and four primary caregivers (185 females [91%], 19 males [9%]) of children with CP aged 4 to 12 years (mean 8 y 4 mo [SD 2.51]; 112 males [55%], 92 females [46%], Gross Motor Function Classification System level I=18%, II=28%, III=14%, IV=11%, V=28%) provided demographic data and completed the CP QOL-Child, CHQ, and KIDSCREEN-10. Fifty-four children with CP aged 9 to 12 years completed the CP QOL-Child and KIDSCREEN-10. RESULTS: The KIDSCREEN-10 and CP QOL-Child were developed to measure general HRQOL and CP specific QOL respectively, whereas the CHQ was developed to measure functional health and well-being. In terms of internal consistency, KIDSCREEN-10 (Cronbach's alpha=0.86) and CP QOL-Child (0.74-0.91) outperformed the CHQ (0.18-0.96). In terms of validity, all instruments were moderately correlated. Floor and ceiling effects, although minimal or not evident for KIDSCREEN-10 and CP QOL-Child (1 4.9%), were apparent for CHQ (0.5-62.9%). INTERPRETATION: Conceptually and psychometrically, KIDSCREEN-10 and CP QOL-Child performed more strongly than the CHQ, for children with CP. The choice between these two instruments will depend on the questions posed and outcomes sought by the researcher or clinician. PMID- 19549194 TI - 'Natalizumab in paediatric multiple sclerosis and service implication'. PMID- 19549195 TI - The vexed issue of cerebral palsy and breech presentation. PMID- 19549196 TI - Aiming to improve the health care of people with cerebral palsy worldwide: a report of an International Society for Prosthetics and Orthotics conference. PMID- 19549197 TI - Sibling sex ratio of individuals diagnosed with autism spectrum disorder as children. AB - AIM: To study the sex ratio (proportion of males) in siblings of individuals diagnosed with autism spectrum disorders (ASDs) as children. METHOD: In the current study, we extended previous studies dealing with the androgen theory of autism and examined sex ratios in the siblings of 326 individuals with ASD (245 males, 81 females) who had been consecutively assessed at two Danish university clinics of child psychiatry during the 25-year period from 1960 to 1985. RESULTS: Among the 513 siblings, 300 were males and 213 females. This yields a sex ratio of 0.585, which is significantly higher than the Danish live-birth sex ratio over the same period (0.514, p=0.001). The sibling sex ratio was not associated with the IQ in the autistic probands. INTERPRETATION: Our findings suggest a potential indirect confirmation of the androgen theory of autism. PMID- 19549198 TI - Participation in home, extracurricular, and community activities among children and young people with cerebral palsy. AB - AIM: Participation in home, extracurricular, and community activities is a desired outcome of rehabilitation services for children and young people with cerebral palsy (CP). The purpose of this study was to investigate the effect of age and gross motor function on participation among children and young people with CP. METHOD: Five hundred participants (277 males, 223 females) were grouped by age and Gross Motor Function Classification System (GMFCS) level. There were 291 children aged 6 to 12 years and 209 young people aged 13 to 21 years. There were 128 participants in GMFCS level I, 220 in levels II/III, and 152 in levels IV/V. Participants completed the Children's Assessment of Participation and Enjoyment to measure number of activities (diversity) and how often they were performed (intensity) in the past 4 months. RESULTS: Children had higher overall participation diversity and intensity than young people (p<0.001). Children and young people in GMFCS level I had the highest overall participation, followed by children and young people in levels II/III and IV/V. Children had higher participation in recreational (p<0.001) but not formal (such as team sports or clubs) or physical activities. Children (p<0.01) and young people (p<0.001) in level I had the highest participation in physical activities; diversity and intensity were generally low. INTERPRETATION: The findings provide evidence of the effect of age and gross motor function on participation of children and young people with CP. Low participation in physical activities may have implications for fitness and health, especially for children and young people in GMFCS levels IV and V. PMID- 19549199 TI - Stability of parent-reported manual ability and gross motor function classification of cerebral palsy. PMID- 19549200 TI - Does parent report measure performance? A study of the construct validity of the Functional Mobility Scale. AB - AIM: Parental report is often relied on to measure performance of activities in children with cerebral palsy (CP). This study examined whether the Functional Mobility Scale (FMS) accurately reflects performance of mobility in children with CP. METHOD: Eighteen children with spastic CP (11 males, seven females; mean age 12 y 8 mo, SD 2 y 8 mo, range 8-17 y) were recruited from a special development school. Children were in Gross Motor Function Classification System (GMFCS) levels II (n=5), III (n=4), or IV (n=9), and had quadriplegia (n=9), diplegia (n=7), or hemiplegia (n=2). The children's mobility was observed directly around and outside the home and at school and their mobility methods were recorded. The parent's FMS rating was obtained on the telephone by a physiotherapist. Agreement between direct observation and the FMS rating was examined using quadratic weighted kappa (kappa) statistics. RESULTS: Agreement between direct observation and the FMS was as follows: FMS 5m kappa=0.71, 45%; FMS 50m kappa=0.76, 94%; FMS 500m kappa=0.74, 95%. Differences in the range and number of mobility methods were observed by GMFCS level across environmental settings. INTERPRETATION: Substantial agreement was found between FMS ratings and direct observation, particularly over longer distances, providing evidence of the validity of the FMS as a measure of performance in children with CP. PMID- 19549201 TI - Differentiating attention deficits in children with fetal alcohol spectrum disorder or attention-deficit-hyperactivity disorder. AB - AIM: The attention and inhibition problems found in children with attention deficit-hyperactivity disorder (ADHD) are also common in children with fetal alcohol spectrum disorders (FASDs). Attempts to distinguish ADHD from FASDs in terms of these deficits are rare and were pursued in this study. METHOD: A total of 116 children (47 with ADHD, 31 males, 16 females; 30 with FASDs, 17 males, 13 females; and 39 comparison children, 20 males, 19 females) participated. The mean age was 9 years 4 months (SD 1y 8mo) in the ADHD groups, 8 years 10 months (SD 1y 2mo) in the FASD group, and 9 years 1 month (SD 1y 1mo) in the comparison group. Sustained attention was tested with a slow event rate continuous performance task (CPT). Inhibitory control was tested with both a slow and fast event rate Go/No Go task. RESULTS: On the CPT task, children with ADHD, combined type (ADHD-C), ADHD, primarily inattentive type (ADHD-PI), and FASDs showed greater declines in task performance as a function of time than comparison children, suggesting sustained attention problems in all clinical groups. Children's Go/No-Go performance was event-rate dependent, with the ADHD-C group being affected in the slow condition and the ADHD-PI and FASD groups having problems with the fast condition. INTERPRETATION: Children with ADHD-C are typically impaired in handling understimulation, while children with FASDs may have problems with overstimulation. The dissociation in responsivity to event rate between groups may have significant differential diagnostic value. PMID- 19549202 TI - Essential fatty acids and attention-deficit-hyperactivity disorder: a systematic review. AB - AIM: Essential fatty acids (EFAs), also known as omega-3 and omega-6 fatty acids, have been claimed to have beneficial effects as a treatment for attention-deficit hyperactivity disorder (ADHD). Animal experiments have provided information about the role of EFA in the brain, and several mechanisms of EFA activity are well known. The current review provides an updated, systematic overview of the theory and use of EFA in ADHD. METHOD: Clinical studies and review papers of EFA blood levels and EFA supplementation trials in children with ADHD were researched in the Medline PubMed database. Additional studies were found from the references of these reports. RESULTS: Children with ADHD present lower levels of blood EFAs, and open-label EFA supplementation trials in ADHD raise EFA blood levels and improve symptoms of ADHD. Randomized controlled trials, however, have generally been unsuccessful in demonstrating any behavioural treatment effects. INTERPRETATION: Current findings do not support the use of EFA supplements as a primary or supplementary treatment for children with ADHD. PMID- 19549203 TI - Degos disease: a new simulator of non-accidental injury. AB - Recent high-profile cases have made paediatricians very aware of the serious implications of either missing or wrongly diagnosing non-accidental injury. Subdural fluid collections in non-mobile infants usually represent haemorrhage caused by non-accidental injury. We report a 6-month-old male who presented to the Accident and Emergency Department of Birmingham Heartlands Hospital with bilateral subdural fluid collections and skin ulcers resembling cigarette burns. Non-accidental injury was considered to be the most likely diagnosis. However, while under observation in hospital, the child's neurological condition deteriorated with progressive cerebral infarctions, and serial photographs of the skin lesions showed failure to heal. The revised diagnosis, confirmed histologically, was Degos disease, an extremely rare and often fatal occlusive vasculopathy. The child was treated palliatively and died 8 weeks after presentation. This report informs doctors of a new simulator of non-accidental injury to be considered in infants with otherwise unexplained subdural fluid collections. PMID- 19549205 TI - Muscle deformity in cerebral palsy: reduced use, overuse, or both? PMID- 19549204 TI - Congenital perisylvian dysfunction - is it a spectrum? AB - AIM: This study examines the overlap between children with bulbar cerebral palsy (Worster-Drought syndrome [WDS]) and perisylvian polymicrogyria. METHOD: A total of 121 children (81 males, 40 females; mean age 5y 5mo, SD 3y 6mo; age range 1mo 15y 4mo) were studied using retrospective clinical data and magnetic resonance imaging. In all, 70 children had WDS with normal perisylvian imaging, 31 had congenital bilateral perisylvian polymicrogyria (CBPP), and 20 had congenital unilateral perisylvian polymicrogyria (CUPP). RESULTS: All groups shared aetiological markers (male sex, congenital contractures, low familial incidence, excess antenatal events). There was a common phenotype of pseudobulbar palsy with mild limb pyramidal signs in all children with WDS, 90% of those with CBPP, and one-third of those with CUPP, often also associated with learning disability*, epilepsy, and behavioural difficulties. A further 15% of children with CUPP acquired this phenotype through an epileptic encephalopathy. Pseudobulbar palsy rather than polymicrogyria was more predictive of additional impairments other than epilepsy. INTERPRETATION: We propose that congenital perisylvian dysfunction is a spectrum encompassing the WDS phenotype and perisylvian polymicrogyria imaging abnormalities. As with other prenatal brain abnormalities, there is not necessarily concordance between imaging and clinical findings, although the phenotype is often more severe to manifest imaging abnormality. Clinical phenotype is the best indicator of prognosis. Epileptic encephalopathy can cause an acquired form of perisylvian dysfunction where there is. PMID- 19549206 TI - Global developmental delay and its relationship to cognitive skills. PMID- 19549207 TI - The assessment of minor neurological dysfunction in infancy using the Touwen Infant Neurological Examination: strengths and limitations. AB - AIM: Little is known of minor neurological dysfunction (MND) in infancy. This study aimed to evaluate the inter-assessor reliability of the assessment of MND with the Touwen Infant Neurological Examination (TINE) and the construct and predictive validity of MND in infancy. METHOD: Inter-assessor agreement was determined in a sample of 40 infants (24 males, 16 females) aged 3 to 12 months (25 born at term: gestational age 37-41wks, median 39; and 15 born preterm, gestational age 24-35wks, median 32). Thirty typically developing term infants (18 males, 12 females; gestational age 37-42wks, median 40) and 59 preterm infants (34 males, 25 females) born at <35 weeks' gestation (gestational age 25 34wks, median 29) participated in the validity study. They were neurologically assessed with the TINE at the corrected ages of 4, 6, 10, and 12 months and with the Hempel assessment at 18 months. RESULTS: The findings indicated that MND can be assessed reliably (inter-assessor agreement: kappa=0.83). MND during infancy was related to prenatal, perinatal, and social factors, and in particular to preterm birth. Neurological condition during infancy was prone to change, but was related to neurological condition at 18 months at all ages tested. INTERPRETATION: We conclude that MND can be determined reliably in infancy. Important considerations in the construct of MND in infancy are its relation to prenatal and perinatal factors, its limited stability, and its moderate predictive value. PMID- 19549208 TI - Prevalence and outcome of esophagogastric anastomotic leak after esophagectomy in a UK regional cancer network. AB - The aim of this study was to determine the contemporary prevalence, outcome, and survival after esophagogastric anastomotic leakage (EGAL) following esophagectomy by a regional upper gastrointestinal cancer network and to investigate etiological factors. Two hundred forty consecutive patients underwent esophagectomy over a 10-year period (median age 61 [31-79] years, 147 transthoracic and 93 transhiatal esophagectomy, 105 neoadjuvant chemotherapy, 49 chemoradiotherapy). The primary outcome measures were the development of EGAL and survival. Twenty patients developed EGAL (8.3%, 15 managed conservatively, 5 reoperation). Overall operative mortality was 2% (5 patients in total, 1 after EGAL). Median, 1 and 2-year survival was 22 months, 73% and 50%, in patients after EGAL, compared with 31 months, 80% and 56%, in patients who did not suffer EGAL (P= 0.314). On multivariate analysis, low body mass indices (hazard ratio [HR] 0.29, 95% confidence interval [CI] 0.11-0.79, P= 0.016), individual surgeon (HR 1.21, 95% CI 1.02-1.43, P= 0.02), and neoadjuvant chemotherapy (HR 3.28, 95% CI 1.16-9.22, P= 0.024) were significantly associated with the development of EGAL. EGAL following esophagectomy remained common, but associated mortality was less common than reported in earlier Western series and long-term survival was unaffected. PMID- 19549209 TI - Endoscopic classification for reflux pharyngolaryngitis. AB - The quality of life in patients who have undergone surgery for esophageal cancer is frequently disturbed by postoperative gastroesophageal reflux disease or pharyngolaryngeal reflux disease. Recently, there have been many reports on gastroesophageal reflux disease after esophagectomy, and only a few on pharyngolaryngeal reflux disease. There is not yet any convenient endoscopic classification of reflux pharyngolaryngitis. We designed a new classification for reflux pharyngolaryngitis based on endoscopic findings. Our new classification consists of the five grades from 0 to IV based on (i) the extent and severity of erythema and/or edema in the pharynx and the larynx, and (ii) the extent and severity of granulation or scarring stenosis in the vocal cords. Ninety-three patients after cervical esophagogastrostomy after esophagectomy (the CEG group) and 28 patients after intrathoracic esophagogastrostomy (the TEG group) were reviewed in this study. We investigated the relation between the severity of reflux pharyngolaryngitis and clinical symptoms in these patients, and the correlation between this new classification of reflux pharyngolaryngitis and the Los Angeles classification of reflux esophagitis. Reflux esophagitis was more severe in the TEG group than in the CEG group, while there was no difference in the grading of reflux pharyngolaryngitis between the two groups. The pharyngolaryngeal symptoms and F-scale scores were not correlated with the severity of reflux pharyngolaryngitis in each group. The grading of reflux pharyngolaryngitis and that of reflux esophagitis was correlated in each group (P<0.001 in the CEG group and P=0.002 in the TEG group). We proposed a new endoscopic classification of reflux pharyngolaryngitis. The new classification of reflux pharyngolaryngitis correlated fairly well with the Los Angeles classification of reflux esophagitis, although this classification did not correlate with the clinical symptoms in patients who underwent esophagectomy. Follow-up attention including upper endoscopy should be paid to reflux pharyngolaryngitis in patients after esophagogastrostomy as well as reflux esophagitis, because there is often a lack in symptoms regardless of high incidence of pharyngolaryngitis. PMID- 19549210 TI - Protective effects of glycoursodeoxycholic acid in Barrett's esophagus cells. AB - Barrett's esophagus (BE) is a premalignant condition associated with the development of esophageal adenocarcinoma (EAC). Previous studies have implicated hydrophobic bile acids and gastric acid in BE and EAC pathogenesis. In this study, we tested the hypothesis that DNA damage, cytotoxicity and oxidative stress induced by bile acids and gastric acid can be attenuated by the cytoprotective, hydrophilic bile acid glycoursodeoxycholic acid (GUDCA). Non dysplastic BE cells were exposed for 10 min to pH 4 and/or bile acid cocktail or to pH 4 and a modified cocktail consisting of a mixture of bile acids and GUDCA. DNA damage was evaluated by the comet assay; cell viability and proliferation were measured by trypan blue staining and the MTS assay; reactive oxygen species (ROS) were measured using hydroethidium staining; oxidative DNA/RNA damage was detected by immunostaining with antibody against 8-OH-dG; thiol levels were measured by 5-chloromethylfluorescein diacetate (CMFDA) staining; and the expression of antioxidant proteins was evaluated by western blotting. DNA damage and oxidative stress were significantly increased, while thiol levels were decreased in BE cells treated with pH 4 and bile acid cocktail compared with cells treated with pH 4 alone or untreated cells. Bile acids and low pH also significantly decreased cell proliferation. Expression of the antioxidant enzymes, MnSOD and CuZnSOD, was elevated in the cells treated with bile acids and low pH. When GUDCA was included in the medium, all these effects of pH 4 and bile acids were markedly reduced. In conclusion, treatment of BE cells with acidified medium and a bile acid cocktail at physiologically relevant concentrations induces DNA damage, cytotoxicity, and ROS. The cytoprotective bile acid, GUDCA, inhibits these deleterious effects by inhibiting oxidative stress. PMID- 19549211 TI - Effectiveness of voice therapy in reflux-related voice disorders. AB - Gastroesophageal reflux (GER) with laryngopharyngeal reflux plays a significant role in voice disorders. A significant proportion of patients attending ear, nose, and throat clinics with voice disorders may have gastroesophageal reflux disease (GERD). There is no controlled study of the effect of voice therapy on GERD. We assessed the effect of voice therapy in patients with dysphonia and GERD. Thirty-two patients with dysphonia and GERD underwent indirect laryngoscopy and voice analysis. Esophageal and laryngeal symptoms were assessed using the reflux symptom index (RSI). At endoscopy, esophagitis was graded according to Los Angeles classification. Patients were randomized to receive either voice therapy and omeprazole (20 mg bid) (n=16, mean [SD] age 36.1 [9.6] y; 5 men; Gp A) or omeprazole alone (n=16, age 31.8 [11.7] y; 9 men; Gp B). During voice analysis, jitter, shimmer, harmonic-to-noise ratio (HNR) and normalized noise energy (NNE) were assessed using the Dr. Speech software (version 4 1998; Tigers DRS, Inc). Hoarseness and breathiness of voice were assessed using a perceptual rating scale of 0-3. Parameters were reassessed after 6 weeks, and analyzed using parametric or nonparametric tests as applicable. In Group A, 9 patients had Grade A, 3 had Grade B, and 1 had Grade C esophagitis; 3 had normal study. In Group B, 8 patients had Grade A, 2 had Grade B esophagitis, and 6 had normal study. Baseline findings: median RSI scores were comparable (Group A 20.0 [range 14-27], Group B 19.0 [15-24]). Median rating was 2.0 for hoarseness and breathiness for both groups. Values in Groups A and B for jitter 0.5 (0.6) versus 0.5 (0.8), shimmer 3.1 (2.5) versus 2.8 (2.0), HNR 23.0 (5.6) versus 23.1 (4.2), and NNE -7.3 (3.2) versus -7.2 (3.4) were similar. Post-therapy values for Groups A and B: RSI scores were 9.0 (5-13; P<0.01 as compared with baseline) and 13.0 (10-17; P<0.01), respectively. Ratings for hoarseness and breathiness were 0.5 (P<0.01) and 1.0 (P<0.01) and 2.0. Values for jitter were 0.2 (0.0; P=0.02) versus 0.4 (0.7), shimmer 1.3 (0.7; P<0.01) versus 2.3 (1.2), HNR 26.7 (2.3; P<0.01) versus 23.7 (3.2), and NNE -12.3 (3.0, P<0.01) versus -9.2 (3.4; P<0.01). Improvement in the voice therapy group was significantly better than in patients who received omeprazole alone. Dysphonia is a significant problem in GER. Treatment for GER improves dysphonia, but in addition, voice therapy enhances the improvement. PMID- 19549212 TI - Lgr5, an intestinal stem cell marker, is abnormally expressed in Barrett's esophagus and esophageal adenocarcinoma. AB - Lgr5 (leucine-rich-repeat-containing G-protein-coupled receptor 5), a recently discovered intestinal stem cell marker, is expressed in premalignant lesions including Barrett's esophagus (BE) and cancers including colon cancer, ovarian cancer, and hepatocellular carcinoma. It was also recently found to be expressed in tumor spheres prepared from colon cancer, suggesting that it will likely serve as a cancer stem cell marker. We sought to examine Lgr5 as a biomarker in BE associated neoplasia. Using standard immunohistochemistry, we performed immunostaining on 81 esophageal specimens (53 biopsy specimens and 28 surgical resections) representing BE, BE-associated dysplasia, and esophageal adenocarcinoma (EAC). Each immunostain was scored based on intensity of immunostaining and percentage of positive cells. For 24 EAC cases, survival analysis was performed with expression scores and other clinicopathological variables. We found that Lgr5 expression was detected in 70% of BE cases and between 90 and 100% of advanced dysplastic lesions and EAC. The intensity of expression was significantly higher in high-grade dysplasia and EAC than BE. In EAC, high Lgr5 expression scores (> or = 5) were associated with worse survival, independent of stage, age, and neoadjuvant/adjuvant therapy (P = 0.03). Our findings suggest that Lgr5 has potential utility as a biomarker for BE-associated dysplasia and EAC. PMID- 19549215 TI - The greening of public lands grazing in the southwestern U.S.A. PMID- 19549213 TI - Immediate implants at fresh extraction sockets: bone healing in four different implant systems. AB - OBJECTIVES: To describe the differences in bone healing, when placing four different implant systems in fresh extraction sockets. MATERIAL AND METHODS: Eight beagle dogs received implants randomly installed into the distal socket of three P3 and four P4. Four-implant systems were evaluated. Each animal provided four test implant sites. All animals were sacrificed at 6 weeks after implant placement, providing specimens for histo-morphometric analysis of bone to implant contact (BIC), bone area, new bone formation, as well as histometric measurements of the ridge alterations. RESULTS: No statistically significant difference was observed among the four-implant systems. The mean BIC % ranged between 58.5% and 72.1%. Bone modelling of the buccal plate was marked and amounted approximately to 2.5 mm, independently of the system used. CONCLUSION: This study failed to demonstrate differences in the healing pattern after 6 weeks when placing four different implant systems in fresh extraction sockets. In spite of achieving predictable osteointegration with the four implants studied, the occurrence of buccal bone resorption may limit the use of this surgical approach. PMID- 19549216 TI - Home gardening for tropical biodiversity conservation. PMID- 19549217 TI - Conservation of the unique neotropical vascular flora of the Guayana Highlands in the face of global warming. PMID- 19549218 TI - Overestimation of fire risk in the northern spotted owl recovery plan. AB - The U.S. Fish and Wildlife Service's recent recovery plan for one of the most carefully watched threatened species worldwide, the Northern Spotted Owl (Strix occidentalis caurina), recommended a major departure in conservation strategies in the northwestern United States. Due to concern about fire, the plan would switch from a reserve to a no-reserve strategy in up to 52% of the owl's range. Fuel treatments (e.g., thinning) at regular intervals also would occur on up to 65-70% of dry forests in this area. Estimations of fire risk, however, were based on less than a decade of data and an anecdotal assessment of a single, large fire. We found that decadal data are inherently too short, given infrequent large fires, to accurately predict fire risk and trends. Rates of high-severity fire, based on remote-sensing data, are far lower than reported in the plan and in comparison with the rate of old-forest recruitment. In addition, over a 22-year period, there has been no increase in the proportion of high-severity fire. Our findings refute the key conclusions of the plan that are the basis for major changes in conservation strategies for the Spotted Owl. The best available science is needed to address these strategies in an adaptive-management framework. From the standpoint of fire risk, there appears to be ample time for research on fire and proposed treatment effects on Spotted Owls before designing extensive management actions or eliminating reserves. PMID- 19549219 TI - A review of climate-change adaptation strategies for wildlife management and biodiversity conservation. AB - The scientific literature contains numerous descriptions of observed and potential effects of global climate change on species and ecosystems. In response to anticipated effects of climate change, conservation organizations and government agencies are developing "adaptation strategies" to facilitate the adjustment of human society and ecological systems to altered climate regimes. We reviewed the literature and climate-change adaptation plans that have been developed in United States, Canada, England, Mexico, and South Africa and found 16 general adaptation strategies that relate directly to the conservation of biological diversity. These strategies can be grouped into four broad categories: land and water protection and management; direct species management; monitoring and planning; and law and policy. Tools for implementing these strategies are similar or identical to those already in use by conservationists worldwide (land and water conservation, ecological restoration, agrienvironment schemes, species translocation, captive propagation, monitoring, natural resource planning, and legislation/regulation). Although our review indicates natural resource managers already have many tools that can be used to address climate-change effects, managers will likely need to apply these tools in novel and innovative ways to meet the unprecedented challenges posed by climate change. PMID- 19549220 TI - Long-term changes in dynamic cerebral autoregulation: a 10 years follow up study. AB - This study takes a novel approach to describing time-related changes in dynamic cerebral autoregulation (dCA). It is well-recognized that dCA exhibits both intra and inter- subject variability, and this study seeks to characterize the extent to which intra-subject variability occurs after a significant period of time by studying the same subjects 10 years apart, thus eliminating inter-subject variability as a source of error. Ten healthy subjects were identified in 1998 and followed up in 2008. On each visit they underwent simultaneous recordings of right middle cerebral artery cerebral blood flow velocity (RMCA CBFV), blood pressure and heart rate. Data were analysed in the frequency domain using transfer function analysis and in the time domain using CBFV step response, from which the autoregulatory index (ARI) was calculated. Ten subjects of mean age 35.5 (range 24-51) years in 1998 (seven male) were studied. There was a significant fall in ARI from 1998-2008 (DeltaARI = 1.1, P = 0.021), along with a significant rise in coherence in 2008 (at 0.05 Hz, P = 0.018). Difference in mean step response between 1998 and 2008 was also significant (P = 0.045). This is the first study to assess dCA in the same subjects 10 years apart, providing a novel opportunity to assess intra-subject variation in dCA after a long time period has elapsed. A fall in frequency and time domain parameters was observed. This is important, and needs to be considered in future studies assessing long-term changes in dCA, particularly given the body evidence which suggests that dCA is unaffected by ageing. PMID- 19549221 TI - Caspofungin for the treatment of candidaemia in patients with haematological malignancies. AB - This study was prospectively conducted in 11 haematology divisions over a 2-year period to evaluate the efficacy of caspofungin in 24 neutropenic patients with haematological malignancies (HM) and candidaemia. These patients had received chemotherapy for HM and were neutropenic (PNN < 0.5 x 10(9)/L) for a median of 12 days (2-41) before candidaemia. The patients received caspofungin for a median duration of 12 days (range 6-26), obtaining a favourable overall response of 58%. At 30 days, 11 patients had died (46%); candidaemia was responsible for mortality in six patients (25%). These results suggest that treatment of candidaemia with caspofungin in neutropenic HM was efficacious, as it is in non-haematological subgroups. PMID- 19549222 TI - The epidemiology of invasive Streptococcus pneumoniae infections in onco haematology and haematopoietic stem cell transplant patients in France. Are the serotypes covered by the available anti-pneumococcal vaccines? AB - The pneumococcal antigens contained in the polysaccharide (PPV23) and conjugate (7-valent, PCV7; 13-valent, PCV13) vaccines have been chosen since they represent the serotypes that more frequently cause invasive pneumococcal disease. Whether these vaccines cover the serotypes most frequently isolated in haematology patients is unclear. The serotype distribution among Streptococcus pneumoniae in 25 consecutive pneumococcal infections that occurred over the last 3 years in two French haematology departments was investigated. The pneumococcal vaccines PCV7, PCV13 and PPV23 were found to cover 76, 84 and 92%, respectively, of the serotypes found. PMID- 19549224 TI - Genetic analysis of four human rabies cases reported in Turkey between 2002 and 2006. AB - Rabies remains endemic in many regions of Turkey. As a consequence, humans are at risk of this fatal disease through encounters with rabid animals. The present study describes four recent cases of rabies in humans. Subsequent phylogenetic analysis of the rabies virus isolates obtained from each case demonstrates the distinct geographical distribution of rabies virus variants within Turkey. The study suggests that rabies virus translocation has occurred across Turkey and might be the source of the emergence of a genetically similar variant in the Golan Heights region on the Israeli/Syrian border in 2004. PMID- 19549225 TI - Macular haemorrhage after repetitive roller coaster riding. PMID- 19549223 TI - Population-based surveillance for scedosporiosis in Australia: epidemiology, disease manifestations and emergence of Scedosporium aurantiacum infection. AB - Australia-wide population-based surveillance for scedosporiosis identified 180 cases, with 118 (65.6%) cases of colonization and 62 (34.4%) cases of infection. Predisposing factors for isolation of Scedosporium spp. included chronic lung disease in 37.8% and malignancy in 21.7% of cases. Predictors of invasive disease (n=62) included haematological stem cell transplantation (n=7), leukaemia (n=16) and diabetes mellitus (n=8). Of 183 phenotypically-speciated isolates, 75 (41%) were Scedosporium prolificans (risk factors: haematologic cancer (n=17), neutropaenia (n=14)) and 108 (59%) had Scedosporium apiospermum/Pseudallescheria boydii phenotype [risk factor: diabetes (n=15)]. Scedosporium prolificans (p 0.01) and leukaemia (p 0.03) independently predicted death. Epidemiological and antifungal susceptibility profiles of Scedosporium aurantiacum (prevalence>or=15.8%) and S. apiospermum were similar. No patient with S. aurantiacum infection (n=6) died. This is the first description of clinical features associated with S. aurantiacum. PMID- 19549226 TI - Refraction, intraocular pressure and anterior chamber depth changes after Nd:YAG laser treatment for posterior capsular opacification in pseudophakic eyes. AB - PURPOSE: The aim was to investigate the effect of Nd:YAG capsulotomy on refraction, intraocular pressure and anterior chamber depth changes and complications of Nd:YAG laser treatment for posterior capsular opacification in pseudophakic eyes. METHODS: Our study includes 26 eyes (23 patients) with posterior capsular opacification after uncomplicated phacoemulsification surgery and intraocular lens implantation. Complete ocular examinations were performed for all patients. The visual acuity, intraocular pressure and anterior chamber depth measurements were obtained in all examinations. Nd:YAG capsulotomy was measured in all patients. Eyes received one drop of aproclonidine 0.5 % before and immediately after YAG laser capsulotomy. Data were analysed statistically. RESULTS: Mean patient age was 53.73 +/- 13.53 years. Before Nd:YAG capsulotomy mean anterior chamber depth was 4.03 +/- 0.58 mm and in the first day after capsulotomy the mean value was 4.02 +/- 0.46 mm. Mean spherical equivalent refraction before laser treatment was -0.52 D and on the first day after laser treatment was -0.49 D. An improvement in visual acuity was achieved in all cases. Before Nd:YAG capsulotomy mean visual acuity was 0.38 +/- 0.13 and on the first day after capsulotomy, the mean value was 0.93 +/- 0.11, the difference of which was statistically significant. There were no statistically significant differences between the anterior chamber depth and intraocular pressure measurements before laser capsulotomy and on the first day, first month and third month after laser. CONCLUSION: Nd:YAG laser capsulotomy is an effective and safe method of treatment of posterior capsular opacification. PMID- 19549227 TI - Keratometric alterations following the 25-gauge transconjunctival sutureless pars plana vitrectomy versus the conventional pars plana vitrectomy. AB - PURPOSE: To assess the alterations in keratometric astigmatism following the 25 gauge transconjunctival sutureless pars plana vitrectomy versus the conventional pars plana vitrectomy. METHODS: Sixteen consecutive patients were enrolled into the study. Conventional vitrectomy was applied to eight of the cases and 25-gauge transconjunctival sutureless vitrectomy was performed in eight patients. Keratometry was performed before and after the surgery. RESULTS: In the 25-gauge transconjunctival sutureless pars plana vitrectomy group, statistically significant changes were not observed in the corneal curvature in any post operative follow-up measurement (p > 0.05); whereas in the conventional pars plana vitrectomy group, statistically significant changes were observed in the first postoperative day (p = 0.01) and first postoperative month (p = 0.03). We noted that these changes returned to baseline in three months (p = 0.26). CONCLUSION: Both 25-gauge transconjunctival sutureless and conventional pars plana vitrectomy are effective surgical modalities for selected diseases of the posterior segment. Surgical procedures are critical for the visual rehabilitation of the patients. The post-operative corneal astigmatism of the vitrectomised eyes can be accurately determined at least two months post-operatively. PMID- 19549228 TI - Anaphylaxis with use of eye-drops containing benzalkonium chloride preservative. PMID- 19549229 TI - Comparison of the clinical features of recurrent aphthous stomatitis and Behcet's disease. AB - BACKGROUND; Recurrent aphthous stomatitis (RAS) presents a diagnostic problem in Behcet's disease (BD), particularly when it occurs as the only or earliest feature of the disease. To date, there have been only a few reports studying the differences in characteristics between RAS and BD. AIM: To examine the clinical differences between RAS and BD using a large group of patients. METHODS: A retrospective review was carried out, analysing demographic data, the clinical features of the oral ulcer, and the major and minor symptoms of BD of 1643 patients with RAS and 3527 patients with BD presenting from 1995 to 2001. RESULTS: BD had a greater female predominance, and major oral ulcers were significantly more common in BD than in RAS (P < 0.001). Involvement of multiple sites was also more common in BD than in RAS, and the menstrual cycle had more influence on oral ulcers in patients with BD (P < 0.001). Minor symptoms such as articular, neurological and vascular symptoms and epididymitis were also seen more often in BD than in RAS (P < 0.001), and in particularly, patients with BD had a significantly higher frequency of articular symptoms than did patients with RAS (P < 0.001). CONCLUSION: These findings may provide guidelines for the clinical differentiation between RAS and BD. In addition, patients with multiple major aphthae, particularly with articular symptoms, should be closely followed up for the development of BD, and the possibility of other diseases such as ankylosing spondylitis and Crohn's disease should also be considered. PMID- 19549230 TI - Solar urticaria successfully treated with intravenous immunoglobulin. AB - Idiopathic solar urticaria (SU) is a rare, debilitating photodermatosis, which may be difficult to treat. First-line treatment with antihistamines is effective in mild cases, but remission after phototherapeutic induction of tolerance is often short-lived. Other treatment options include plasma exchange, photopheresis and cyclosporin. We present two cases of severe, idiopathic SU, which were resistant to conventional treatment. Both patients achieved remission after administration of intravenous immunoglobulin (IVIg) and have remained in remission at 13 months and 4 years, respectively. There are only two case reports of successful treatment of solar urticaria with IVIg. In our experience IVIg given at a total dose of 2 g/kg over several 5-day courses about a month apart is an effective treatment option for severe idiopathic SU. It is also generally safe, even if certainly subject to significant theoretical risks, such as induction of viral infection or anaphylaxis. PMID- 19549231 TI - Spontaneous regression of subcutaneous in-transit malignant melanoma deposits of the lower leg after treatment with the carbon dioxide laser. AB - Primary malignant melanoma (MM) is one of the few cancers that can show spontaneous regression, and although this is well described, the mechanisms behind this are unclear. Spontaneous regression of secondary metastatic deposits of melanoma is rare and there are only a handful of case reports supporting this phenomenon. We report a patient with in-transit MM metastatic disease of the leg, who underwent carbon dioxide (CO(2)) laser ablation, and subsequently developed remission of untreated deposits. We discuss the possible immunological mechanism for this phenomenon. PMID- 19549232 TI - Treatment with carbamazepine and gabapentin of a patient with primary erythermalgia (erythromelalgia) identified to have a mutation in the SCN9A gene, encoding a voltage-gated sodium channel. AB - Primary erythermalgia (erythromelalgia) is a rare autosomal dominant condition characterized by intermittent attacks of erythema, increased skin temperature and severe burning pain in the extremities, in a bilateral symmetrical distribution. Mutations in the SCN9A gene, which encodes a voltage-gated sodium channel have been shown to cause this disease. We report a family identified to have a mutation in the SCN9A gene, in which one severely affected family member has responded to the therapeutic combination of gabapentin and carbamazepine treatment. PMID- 19549233 TI - Angiolymphoid hyperplasia with eosinophilia: good response to photodynamic therapy. AB - Angiolymphoid hyperplasia with eosinophilia (ALHE) is an uncommon benign vascular disease of unknown pathogenesis, that occurs as solitary or multiple nodules or papules. Lesions are located mainly on the head, forehead and neck, and may be persistent and difficult to eradicate. We report a case of ALHE treated with aminolaevulinic acid photodynamic therapy (ALA-PDT). Treatment consisted of two ALA-PDT sessions with a 2-week interval. Clinical evaluation, 8 weeks after treatment, showed marked improvement though not complete regression. The treatment was well tolerated. At follow-up 4 months after treatment, the lesions were stable. We believe that PDT could be an alternative therapeutic approach for ALHE or could be used as a neoadjuvant treatment to reduce lesion size especially where size or site of lesions limits the efficacy or acceptability of other treatments. The lack of cumulative effects allows repeated treatments with ALA PDT, but long-term follow-up is advised for assessment of recurrence. PMID- 19549234 TI - Successful treatment of 'leucotrichotillomania' by hair dyeing. PMID- 19549235 TI - Treatment of facial flushing by topical application of nicotinic acid cream followed by treatment with 595-nm pulsed-dye laser. PMID- 19549236 TI - Hair diagnoses and signs: the use of dermatoscopy. AB - BACKGROUND: Hair-shaft examination is diagnostically useful in a range of adult and paediatric conditions. OBJECTIVE: To evaluate the usefulness of dermatoscopy in hair-shaft microscopy. METHODS: Typical examples of selected conditions from an extensive collection of scalp hair were examined using a dermatoscope and a light microscope with paired cross-polarizing filters. Hair-shaft characteristics were photographed using a digital camera. RESULTS: Dermatoscopy was helpful in detecting tapered hairs, weathering, monilethrix, pediculosis capitis, peripilar casts, 'exclamation-mark' hairs of alopecia areata, bubble hair and pili torti. It was less helpful in pili annulati and unhelpful in detecting 'tiger-tail' banding in trichothiodystrophy. Light microscopy provided greater detail in almost all cases; it was necessary for detection of cuticle changes and added significant information in detecting characteristic features of trichothiodystrophy, pili annulati, bubble hair and pili torti. CONCLUSIONS: Dermatoscopy is most revealing in conditions resulting in gross changes in shaft outline and colour, where reflected light is valuable. It is unhelpful for detection of features within the shaft or at higher levels of resolution. When added to its ability to aid evaluation of scalp surface characteristics, dermatoscopy provides an excellent first-line method of assessment in clinics. In vivo it may aid screening and selection of hairs of greatest diagnostic yield for further assessment. In some instances, it may obviate the need for obtaining hair specimens and have implications for public health screening. Where detailed or cortical hair-shaft features need assessment, transmitted light microscopy remains the standard tool. PMID- 19549237 TI - Subacute cutaneous lupus erythematosus associated with terbinafine. PMID- 19549238 TI - Migraine with aura induced by efalizumab. PMID- 19549239 TI - Resorcinol peels as a possible self-treatment of painful nodules in hidradenitis suppurativa. AB - BACKGROUND: Hidradenitis suppurativa (HS) is a chronic, inflammatory skin disease characterized by abscess formation localized to apocrine sweat gland-bearing skin. The most important factor in patients' overall assessment of disease severity is pain. The duration of abscesses takes days to weeks and are always painful. AIM: To assess the efficacy of self-treatment with topical 15% resorcinol in an open study. METHODS: The case notes of 12 women with stage 1 or 2 HS treated with topical resorcinol and followed up for at least 1 year were reviewed. The patients rated the efficacy of treatment on global maximum pain of nodules and abscesses on a visual analogue scale (VAS) and by self-report of the mean duration (days) of a painful lesion. RESULTS: All patients experienced a significant decrease in pain as assessed by VAS and reported a reduction in mean duration of the painful abscesses. CONCLUSIONS: Topical treatment with 15% resorcinol reduced pain from painful nodules in all patients with HS. Further trials are warranted to confirm these results. PMID- 19549240 TI - Muir-Torre syndrome: extraocular sebaceous carcinoma with adenocarcinoma of colon in a 76-year-old man. PMID- 19549241 TI - Amicrobial pustulosis of the folds associated with Hashimoto's thyroiditis. AB - Neutrophilic dermatoses comprise a wide spectrum of diseases characterized histologically by the presence of an aseptic neutrophilic infiltrate in the skin. The neutrophilic infiltrate may move from the epidermis to the dermis and the subcutis, resulting in distinct clinicopathological conditions. There have been a few reports on a peculiar pustular dermatosis involving the cutaneous flexures and arising in patients with autoimmune diseases or immunological abnormalities. We report a patient who developed amicrobial pustulosis of the folds soon after she was diagnosed with Hashimoto's thyroiditis (HT). To our knowledge, the association of APF with HT is a novel finding that has not previously been described. We believe that this combination is not just coincidental, but may share similar immunopathological mechanisms. PMID- 19549242 TI - An investigation of the association between diet and occurrence of acne: a rational approach from a traditional Chinese medicine perspective. AB - BACKGROUND: The association between diet and acne vulgaris has long been suggested but remains unproven in western medicine. In contrast, the diet-acne relationship is considered important in traditional Chinese medicine (TCM). Aim. To investigate the association between diet and acne, using a TCM approach. METHODS: Using a cross-sectional study of 322 entrants to a university in Hong Kong, China, we examined the participants' clinical severity of acne using the Global Acne Grading System and the participants' yin and yang scores using a quantitative method. We then divided them into two groups, a yin-predominant group (yin-PG) and a yang-predominant group (yang-PG) before the diet-acne relationship was investigated. RESULTS: In total, 82 (25.2%) participants were in the clinical acne group and 240 (74.5%) were in the reference group. There were 155 (48.1%) participants in the yin-PG and 167 (51.9%) in the yang-PG group. No association of diet and acne was found when the participants were considered as a homogenous group. In yin-PG, intake of foods from street stalls (P = 0.04) was significantly associated with a lower incidence of acne. In yang-PG, the intake of desserts (P = 0.04) and fresh fruit juices (P = 0.02) was significantly associated with a higher incidence of acne, whereas the intake of dairy and soy products (P = 0.04) was significantly associated with a lower incidence of acne. CONCLUSIONS: The application of a TCM approach led to the detection of significant associations between diet and the incidence of acne. PMID- 19549243 TI - Behcet's disease and vitiligo in two brothers: coincidence or association? AB - Behcet's disease (BD) and vitiligo are disorders with unknown aetiology. We report on two brothers with ocular Behcet's disease who had advanced unilateral ophthalmic symptoms associated with vitiligo. The two brothers had recurrent oral and genital ulceration, uveitis and white patches on their skin. The most probable hypothesis for the aetiology of BD is that of an autoimmune reaction in genetically predisposed individuals, with vasculitis as the main pathological hallmark. Despite many years of research, the specific causes of vitiligo remain obscure, and the most advanced aetiological hypothesis remains that of autoimmunity. To our knowledge, this is the first reported case within the literature of BD associated with vitiligo. The existence of the two different disorders is noteworthy as they were observed in two brothers during the same period in their lives, with very similar clinical observations. PMID- 19549244 TI - Efficacy of topical tacrolimus 0.3% in clobetasol propionate 0.05% ointment in therapy-resistant cutaneous lupus erythematosus: a cohort study. AB - BACKGROUND: Despite a range of available topical and systemic therapies, treatment of cutaneous lupus erythematosus (CLE) can be challenging. Objectives. To evaluate the efficacy of a specially formulated preparation of tacrolimus 0.3% in clobetasol propionate 0.05% ointment (TCPO) in the treatment of CLE. METHODS: Case notes of 13 patients with treatment-resistant CLE (11 discoid LE, 1 systemic LE and 1 subacute cutaneous LE) who had used twice-daily TCPO (TCPO group) were reviewed. These were compared with five similar patients with resistant CLE who had been given 0.1% tacrolimus ointment alone (TO group). RESULTS: In the TCPO group (mean treatment duration 20 months, range 1-72), a good or excellent response was seen in five and six patients, respectively; one patient showed slight improvement. Telangiectasia and acne were observed in two patients. No systemic side-effects were noted. In the TO group (mean treatment duration 6 months, range 1-24), one patient showed good improvement and two showed slight improvement. CONCLUSION: The results of our small retrospective study suggest that TCPO may be more effective than either 0.1% tacrolimus or clobetasol propionate 0.05% ointment monotherapy in the treatment of recalcitrant CLE. Randomized controlled trials are needed to confirm these preliminary findings. PMID- 19549245 TI - Immune evasion during varicella zoster virus infection of keratinocytes. AB - T cells are sensitized during varicella-zoster virus (VZV) infection and are important for control of viral spread and reactivation. In this report, we show that human keratinocytes infected with VZV inhibited upregulation of major histocompatibility complex (MHC) class I, MHC class II and intercellular adhesion molecule-1 after interferon (IFN)-gamma treatment. The ability of keratinocytes to upregulate MHC class I in response to IFN-alpha, tumour necrosis factor (TNF) alpha and Toll-like receptor (TLR)-3 ligand was also diminished upon VZV infection. VZV-infected keratinocytes treated with IFN-gamma had significantly reduced capacity to stimulate antigen-specific T cells compared with uninfected cells. Interference with IFN-alpha, TNF-alpha, IFN-gamma and TLR-3 signalling in keratinocytes by VZV may contribute to immune evasion of the adaptive immune response. PMID- 19549246 TI - The adipokine zinc-alpha2-glycoprotein (ZAG) is downregulated with fat mass expansion in obesity. AB - INTRODUCTION: Zinc-alpha2-glycoprotein (ZAG) is a novel adipokine, which may act locally to influence adipocyte metabolism. This study assessed the effect of increased adiposity on ZAG expression in adipose tissue in human subjects. The study also examined the association between ZAG and adiponectin expression in human adipose tissue, and whether ZAG modulates adiponectin secretion by human adipocytes. METHODS: Adipose tissue (visceral and subcutaneous) was collected from human subjects with a wide range of BMIs. Human Simpson-Golabi-Behmel syndrome (SGBS) adipocytes were used for in vitro studies. ZAG mRNA levels were quantified by real-time PCR and protein by Western blotting. RESULTS: In human subjects, ZAG mRNA level was negatively correlated with BMI (r = -0.61, P < 0.001, n = 23, visceral; r = -0.6, P < 0.05, n = 14, subcutaneous) and fat mass (r = -0.62, P < 0.01, visceral; r = -0.6, P < 0.05, subcutaneous). Negative associations were also found between ZAG mRNA and insulin resistance parameters including plasma insulin (r = -0.65, P < 0.001, visceral; r = -0.55, P < 0.05, subcutaneous) and homeostasis model of insulin resistance (HOMA-IR) (r = -0.65, P < 0.001, visceral; r = -0.52, P = 0.055, subcutaneous), and C reactive protein (CRP) (r = -0.46, P < 0.05, visceral; r = -0.53, P < 0.05, subcutaneous). However, ZAG mRNA was positively correlated with adiponectin (r = 0.5, P < 0.05, visceral; r = 0.82, P < 0.001, subcutaneous) but negatively associated with leptin mRNA (r = -0.42, P < 0.05, visceral; r = -0.54, P < 0.05, subcutaneous). ZAG secretion by differentiated human adipocytes was abundant. Addition of recombinant ZAG stimulated adiponectin release from human adipocytes. CONCLUSION: ZAG gene expression in adipose tissue is downregulated with increased adiposity and circulating insulin. ZAG mRNA is positively correlated with adiponectin mRNA, and ZAG enhances adiponectin production by human adipocytes. We suggest that ZAG is linked to obesity and obesity-related insulin resistance. PMID- 19549247 TI - Factors determining the remission of microprolactinomas after dopamine agonist withdrawal. AB - BACKGROUND: Withdrawal of dopamine agonist (DA) therapy in the management of microprolactinoma is common practice, but it is unclear which patients are likely to attain long-term remission. OBJECTIVE: To identify predictive factors for long term remission. DESIGN: Prospective cohort study. PATIENTS: Forty subjects (39 female, aged 24-60 years) with microprolactinoma; all had been normoprolactinaemic on DA therapy for at least 2 years [mean duration of therapy 9 years (range 2-27)]. MEASUREMENTS: A pituitary magnetic resonance imaging (MRI) was performed on 36 (90%) subjects before DA withdrawal. Relapse was defined as prolactin greater than 480 mIU/l (22.8 microg/l) on two occasions. RESULTS: Nine out of 40 (22.5%) subjects were normoprolactinaemic 12 months after DA withdrawal. Amongst the relapse group, 24 of 31 subjects (79.4%) had already relapsed at 3 months. Normalization of MRI prior to DA withdrawal (P = 0.0006) and longer duration of DA treatment (P = 0.032) were significant predictors of remission. Age, pre-treatment prolactin, nadir prolactin, previous failure of DA withdrawal, pregnancy, dose and type of DA were not significant predictors of remission. The nine patients who were in remission at 12 months were then followed up for 58.0 +/- 5.8 months; all remained in remission. CONCLUSIONS: As many as 22.5% of subjects with microprolactinoma remained normoprolactinaemic 12 months after DA withdrawal and these subjects stayed in remission for up to 5 years. Significant predictive factors were normalization of MRI prior to discontinuation, and duration of DA treatment. Our findings support intermittent DA withdrawal after a period of normoprolactinaemia, particularly where MRI appearances have normalized. PMID- 19549248 TI - Morbidity in patients on long-term steroid replacement therapy. PMID- 19549249 TI - Short-term overt hypothyroidism induces sympathovagal imbalance in thyroidectomized differentiated thyroid carcinoma patients. AB - CONTEXT: Thyroid hormone impacts on the cardiovascular system. (Subclinical) hyperthyroidism results in sympathovagal imbalance due to decreased vagal tone. However, conflicting data have been reported on the effects of hypothyroidism on the activity of the autonomic nervous system (ANS). In hypothyroidism, both increased and decreased sympathetic activity and increased vagal tone have been found. OBJECTIVE: To study the effects of acute short-term overt hypothyroidism and thyroxine replacement therapy on the ANS by measuring urinary excretion of catecholamines and heart rate variability (HRV). DESIGN: Prospective study. SETTING: University hospital. PATIENTS: We studied 11 patients, previously treated with thyroidectomy for differentiated thyroid carcinoma, during hypothyroidism caused by cessation of thyroxine substitution for 4 weeks and during thyroxine replacement therapy, and 21 matched healthy controls. MAIN OUTCOME MEASURES: The activity of the ANS was assessed by measuring urinary excretion of catecholamines and HRV at rest and during a challenge of the ANS by a mental stress test. RESULTS: Urinary dopamine excretion was significantly lower during hypothyroidism. Although in the patients total variability was unchanged, HRV analysis showed a significantly lower low frequency/high frequency ratio, indicating sympathovagal imbalance with sympathetic withdrawal. The mental stress test in the patients resulted in a significant increase in heart rate to the extent of 16-18%. This response was not different between the hypothyroid state and during thyroxine replacement therapy suggesting that cardiovascular reflexes in these patients remain intact. CONCLUSION: Acute short-term overt hypothyroidism results in sympathovagal imbalance with sympathetic withdrawal, with preservation of the cardiovascular reflexes to (mental) stress. PMID- 19549250 TI - High cortisol levels in hyperglycaemic myocardial infarct patients signify stress hyperglycaemia and predict subsequent normalization of glucose tolerance. AB - CONTEXT: It is unclear if people who develop stress hyperglycaemia have underlying abnormal glucose metabolism, an exaggerated hormonal response to stress, or both. Similarly, it is unknown whether stress hyperglycaemia predicts future glucose intolerance. OBJECTIVE: To determine the relationship between illness severity and plasma cortisol concentration with the degree of hyperglycaemia in subjects experiencing acute myocardial infarction (AMI), and their later glucose metabolic status. PATIENTS AND DESIGN: This analysis comprised 55 patients from the HI-5 Study--a randomized control trial of insulin glucose infusion therapy for AMI patients with hyperglycaemia. MAIN OUTCOME MEASURES: Blood glucose level (BGL) as well as total and free cortisol levels on admission were measured. Patients not previously known to have diabetes were assessed for abnormal glucose metabolism following discharge. RESULTS: Patients with ST segment-elevation myocardial infarction (STEMI) and higher peak creatinine kinase level had a higher mean admission total and free cortisol level. As many as 38.5% of subjects were found to have newly diagnosed glucose intolerance at follow up. After multiple logistic regression, admission BGL was found to be a positive predictor (P = 0.027) whereas total cortisol level (P = 0.01) was a negative predictor for glucose intolerance. CONCLUSIONS: Both the level of hyperglycaemia and cortisol levels on admission are predictive for the subsequent abnormal glucose tolerance development in hyperglycaemic AMI patients. Hyperglycaemia in patients who are more unwell (i.e. higher cortisol) reflects the stressed state rather than underlying glucose intolerance. Conversely, if the patient is less sick (i.e. lower cortisol), hyperglycaemia is more likely to reflect underlying glucose intolerance. PMID- 19549251 TI - Prevalence of clinical remission in patients with sporadic idiopathic hypoparathyroidism. AB - BACKGROUND: Remission of disease activity is a characteristic feature of autoimmune endocrine disorders such as Graves' disease, Addison's disease and occasionally in patients with premature ovarian failure. Autoimmunity is also implicated in sporadic idiopathic hypoparathyroidism (SIH) with clinical remission of disease reported in three cases. OBJECTIVE: To assess the rate of remission in patients with sporadic idiopathic hypoparathyroidism and review the cases reported so far. SUBJECTS AND METHODS: Subjects included 53 patients (M:F, 24:29) with SIH who had been symptomatic for at least 1 year (range 1-31 years). They were treated with calcium and 1-alpha-(OH)D(3)/cholecalciferol therapy and had a mean duration of follow up of 5.0 +/- 3.2 years. Treatment was withdrawn in two stages in the patients who maintained normal levels of serum total calcium during the preceding year of treatment. In stage-1, the dose of therapy was reduced to half and subsequently all treatment was stopped (stage 2) in those patients who maintained normal serum total calcium levels on the reduced dose. Remission of SIH was defined as maintenance of normal serum total (>or=2.12 mmol/l) and ionized calcium, inorganic phosphorus and serum intact parathyroid hormone (iPTH) for at least 3 months after withdrawal of calcium and 1-alpha (OH)D(3)/cholecalciferol therapy. Calcium sensing receptor autoantibodies (CaSRAb) were determined by Western blot. RESULTS: Two of the 53 patients (3.8%) with SIH stayed in remission for 1 year after complete withdrawal of therapy. CaSRAb was absent in both the cases. The clinical features, age at onset and duration of hypocalcaemic symptoms in cases with remission were comparable to those who did no show remission. CONCLUSION: Sporadic idiopathic hypoparathyroidism is not irreversible as is widely believed and spontaneous remission of disease may occur in 3.8% of patients. PMID- 19549252 TI - Expression of autotaxin and acylglycerol kinase in prostate cancer: association with cancer development and progression. AB - Lysophosphatidic acid (LPA) may enhance diverse biologic activities in prostate cancer. This study was conducted to analyze expression levels of LPA-producing enzymes, autotaxin (ATX) and acylglycerol kinase (AGK), in prostate cancer with relevance to clinicopathological parameters. Real-time RT-PCR and western blotting were performed for ATX and AGK in non-neoplastic prostate cells (PrECs and PrSCs) and prostate cancer cell-lines (DU-145, PC-3, LNCaP, and AILNCaP). Immunohistochemical analyses were conducted in tissue specimens of 132 localized prostate cancer patients who underwent radical prostatectomy between 2001 and 2007 (median observation period, 22 months). Both enzymes were negatively expressed in PrECs and PrSCs at mRNA and protein levels. ATX expression was higher than AGK in AILNCaP, DU-145, and PC-3 cell-lines, while AGK was mainly expressed in LNCaP cells. Immunohistochemically, ATX and AGK expressions were negative in non-neoplastic epithelia, while both were weakly expressed in the majority of high-grade intra-epithelial neoplasia (HG-PIN). In cancer foci, ATX and AGK expressions were strong in 49% and 62%, weak in 40% and 32%, and negative in 11% and 6%, respectively. Expressions of both enzymes were significantly correlated with primary Gleason grade of cancer foci (P < 0.0001) and capsular invasion (P = 0.03 and 0.003 respectively). ATX expression was significantly correlated with probability of prostate specific antigen (PSA)-failure after surgery (P < 0.0001). In conclusion, LPA-producing enzymes (ATX and AGK) were frequently expressed in prostate cancer cells and precancerous HG-PIN. In particular, high expression levels of ATX were associated with both malignant potentials and poor outcomes. PMID- 19549253 TI - Downregulation of MSP58 inhibits growth of human colorectal cancer cells via regulation of the cyclin D1-cyclin-dependent kinase 4-p21 pathway. AB - We have investigated the expression and role of the 58-kDa microspherule protein (MSP58) in colorectal carcinoma (CRC). By immuhistochemistry and immunofluorescence, we observed MSP58 in the nucleus and cytoplasm of CRC cells, and found MSP58 to be present in CRC specimens more often than in adjacent non tumor tissues (92.5 vs 36.3%, P < 0.01). The average staining score in adjacent non-tumor tissues was significantly lower than in CRC tissues (2.05 +/- 1.13 vs 5.23 +/- 1.38, P < 0.01). Moreover, MSP58 mRNA and protein appeared to be upregulated in six fresh CRC samples compared to their adjacent non-cancerous tissues. MSP58 expression was also detected in the human CRC-derived cell lines LoVo, CoLo205, HCT116, HT-29, SW620, and SW480. Downregulation of MSP58 inhibited in vitro growth and attenuated tumor growth in animal models by induction of cell cycle arrest, and was associated with reduced levels of cyclin D1, cyclin dependent kinase 4, phosphorylation-Rb (p-Rb), p21, and Retino blastoma (Rb) proteins. These results indicated that MSP58 might play an important role in the carcinogenesis of CRC via regulation of the cyclin D1-cyclin-dependent kinase 4 p21 pathway. PMID- 19549254 TI - Expression of KAI1/CD82 in distant metastases from estrogen receptor-negative breast cancer. AB - The tetraspanin protein superfamily member KAI1 suppresses tumor growth and metastasis in animal models and is downregulated in various human malignancies. In breast cancer, KAI1 is preferentially lost in estrogen receptor (ER)-positive tumors. Interestingly, most ER-negative primary breast cancers retain KAI1 expression. This study aimed to evaluate whether or not KAI1 is downregulated during progression to metastasis of these carcinomas. Expression of KAI1, ER, progesterone receptor, c-ErbB2, and Ki67 was analyzed in tissue microarrays comprising a large collection of distant organ metastases from human breast cancers (n = 92) by immunohistochemistry. Results were compared with a previously characterized set of primary breast tumors (n = 209). Immunoreactivity for KAI1 was observed in one-third of the metastases and was associated with lack of ER expression (P = 0.005). The high frequency of KAI1-positive cases in ER-negative primary tumors was maintained in ER-negative metastases. Expression of KAI1 was also observed in MDA-MB-468 and SK-BR-3, two ER-negative breast cancer cell lines of metastatic origin. Moreover, a reanalysis of independent microarray gene expression data indicated maintenance of KAI1 mRNA expression in metastases from ER-negative breast cancers. Furthermore, in a series of matched pairs of mammary carcinomas and metachronous distant metastases, all metastases from KAI1 positive/ER-negative primary tumors were KAI1-positive as well. Collectively, these findings demonstrate that the expression of KAI1 is maintained during progression to metastasis in a large proportion of ER-negative mammary carcinomas. This has significant implications for the use of KAI1 as a clinical marker and the understanding of the metastatic process in human breast cancer. PMID- 19549255 TI - Narrow band imaging for detecting residual/recurrent cancerous tissue during second transurethral resection of newly diagnosed non-muscle-invasive high-grade bladder cancer. AB - OBJECTIVE: To determine if narrow-band imaging (NBI) can be used to detect high grade cancerous lesions missed with the white light at the time of a second transurethral resection (TUR) of high-grade non-muscle-invasive bladder cancer (NMIBC). PATIENTS AND METHODS: Consecutive patients with newly diagnosed high grade NMIBC were enrolled in a prospective observational study. Patients with incomplete resection or absence of muscle tissue in the specimen were excluded. About 1 month after the first TUR, NBI cold-cup biopsies were taken from areas suspicious for urothelial cancer at the end of an extensive white-light second TUR protocol including: (i) resection of the scar of the primary tumour; (ii) resection of any overt or suspected urothelial lesions; and (iii) six random cold cup biopsies of healthy mucosa. RESULTS: In 2008, 47 consecutive patients were recruited after giving written consent (median age 62 years, range 49-83, 39 men and eight women). Nine patients (19%) had macroscopic or microscopic high-grade NMI urothelial cancer, whereas one was reassessed as having muscle-invasive disease at the white-light second TUR plus the six random biopsies. NBI biopsies were taken in 40 of the 47 patients and detected six more patients with high grade cancerous tissue (13%). In all 16 of the 47 patients (34%) were found to have residual/recurrent cancer using our extensive protocol of second TUR followed by NBI biopsies. CONCLUSIONS: Adding NBI biopsies at the end of an extensive second TUR protocol in patients with newly diagnosed high-grade NMIBC can lead to the identification of patients with otherwise missed high-grade residual/recurrent urothelial carcinoma. PMID- 19549256 TI - Screening for prostate cancer appears to work, but at what cost? PMID- 19549257 TI - Site of positive surgical margins influences biochemical recurrence after radical prostatectomy. AB - OBJECTIVE: To determine whether the number and location of positive surgical margins (PSMs) in radical prostatectomy (RP) surgical specimens affect biochemical recurrence (BCR) rates. PATIENTS AND METHODS: The locations of PSMs were recorded for 1308 consecutive men who underwent RP between October 2000 and December 2006. BCR was defined as three consecutive prostate-specific antigen (PSA) level rises with the peak level >or=0.15 ng/mL. Multivariate regression analyses were used to identify preoperative predictors of PSMs and BCR. The estimated 5-year risk of BCR was calculated using the Kaplan-Meier method. RESULTS: In all, 128 (9.8%) men had one or more PSMs. The mean body mass index, mean preoperative serum PSA level, the distributions of clinical stage and biopsy Gleason scores, and the presence or absence of biopsy perineural invasion were significantly different between men with or with no PSMs. In multivariate analysis, baseline serum PSA level, Gleason score and perineural invasion were independent preoperative predictors of PSMs. The 5-year actuarial BCR rates were dependent on the site of the PSM (P = 0.035) and not the number of PSMs (P = 0.18). The rank order of estimated 5-year BCR rates according to the site of PSMs were base > anterior > posterolateral > apex approximately posterior. CONCLUSIONS: About half of the men with PSMs in the RP surgical specimen in our prospective series did not develop BCR. The risk of BCR was dependent on the site and not the number of PSMs. Adjuvant therapy should be considered in cases with anterior and basilar PSMs due to the very high risk of BCR. PMID- 19549258 TI - In idiopathic calcium oxalate stone-formers, unattached stones show evidence of having originated as attached stones on Randall's plaque. AB - OBJECTIVE: To analyse the structure and composition of unattached stones in idiopathic calcium oxalate (CaOx) stone-formers (ICSF) and compare them to attached stones from the same cohort, to investigate whether there is more than one pathogenic mechanism for stone formation in ICSF. PATIENTS AND METHODS: ICSF undergoing percutaneous nephrolithotomy or ureteroscopy for the treatment of nephrolithiasis gave consent to participate in this study. All accessible renal papillae were endoscopically imaged using a digital endoscope. All stones were removed and determined by the operating surgeon to be attached or unattached to the underlying papilla. Micro-computed tomography (micro-CT), which provides three-dimensional analysis of entire stones, was used to compare the structure and composition of attached and unattached stones. RESULTS: Of 115 stones collected from nine patients (12 renal units), only 25 stones were found not to be attached to renal papillae. Of these 25 stones, four were lost and 12 showed definite morphological evidence of having been attached to tissue, probably having been displaced from papillae during access. For the remaining nine stones, micro-CT analysis showed at least one internal region of calcium phosphate within each of these unattached CaOx stones, i.e. the internal structure of the unattached stones is consistent with their having originated attached to Randall's plaque, and then having become detached but retained in the kidney, with new layers of CaOx eventually covering the original attachment site. CONCLUSIONS; Micro-CT analysis supports the hypothesis that in ICSF, both attached and unattached stones occur as a result of a common pathogenic mechanism, i.e. in this type of stone former, CaOx stones, even those not showing morphology that betrays attachment, all originate attached to interstitial plaque on the renal papilla. PMID- 19549259 TI - The aggressiveness of urinary tract urothelial carcinoma increases with the severity of chronic kidney disease. AB - OBJECTIVE: To assess, in a retrospective cohort, urinary tract urothelial carcinoma (UT-UC) in patients with various stages of chronic kidney disease (CKD) and their clinicopathological features, as patients with end-stage renal disease (ESRD) have a higher incidence of UT-UC, but the relationship between early stages of CKD and characteristics of UT-UC are less well known. PATIENTS AND METHODS: The study included 267 patients with pathologically confirmed UT-UC from January 1994 to December 2006; all had a physical examination (blood pressure), and measurements of laboratory data (serum creatinine, serum haemoglobin) and pathological data. The glomerular filtration rate (GFR) was calculated using the Modification of Diet in Renal Disease equation. Patients were divided into three groups by individual GFR (mL/min), i.e. >60 (no/mild CKD), 30-60 (CKD stage 3) and <30 (CKD stage 4/5). RESULTS: The CKD stages included 81 (30.3%) patients with none/mild CKD, 121 (45.3%) with CKD stage 3 and 65 (24.3%) with CKD stage 4/5. There was a significant and parallel increase in the frequency of UT-UC as CKD severity increased from none/mild CKD to stage 3 (11% vs 55%), and from CKD stage 3 to 4/5 (55% vs 71%; P < 0.05). Pathologically, the frequency of high grade and high T stage UT-UC in patients with CKD stage 3 (90% and 35%, respectively) and CKD stage 4/5 (91% and 29%, respectively) were significantly greater than in the group with none/mild CKD (P < 0.001). Advanced age and more distant metastasis were independent risk factors for patient survival. CONCLUSION: The aggressiveness of UT-UC increased with the severity of CKD, and this might have important clinical consequences. PMID- 19549261 TI - Prostate-sparing cystectomy: long-term oncological results. AB - OBJECTIVE: To analyse the oncological outcome of prostate-sparing cystectomy (PSC). PATIENTS AND METHODS: Between 1994 and 2006, 63 men were treated with PSC after meeting the inclusion criteria (no tumour at the bladder neck, no prostate cancer). The results were compared with patients who had a standard cystoprostatectomy (SC) during the same study period, after matching for clinical and pathological characteristics. RESULTS: The 3- and 5-year disease-specific survival rates were 77% and 66% in the PSC group, and 68% and 64% in the SC group (log-rank, P = 0.6). The local recurrence rate was 7.9% and 16% for the PSC and the SC groups, respectively, and the respective distant recurrence rate was 29% and 33%. Subsequent prostate cancer was detected in 3% in the PSC group. None of these patients died from prostate cancer. In the SC group the final pathology showed that 18% had prostate cancer. CONCLUSION: Local recurrences were not diagnosed more often in the PSC than the SC group. The outcomes of both procedures are comparable with contemporary cystoprostatectomy series. We consider this procedure oncologically safe and offer this to selected patients. However, selection is the key to success, and our results should further be corroborated by the experience of others. PMID- 19549260 TI - Alendronate decreases the fracture risk in patients with prostate cancer on androgen-deprivation therapy and with severe osteopenia or osteoporosis. AB - OBJECTIVE: To evaluate changes in bone mass and fracture risk in patients with prostate cancer on androgen-deprivation therapy (ADT) and with a basal T-score of >-2.0, who were treated with an oral bisphosphonate, as such patients treated with ADT are at increased risk of bone loss and bone fracture. PATIENTS AND METHODS: We selected 61 patients with prostate cancer treated with ADT; 31 were treated with oral alendronate 70 mg once-weekly and a control group of 30 were not. At baseline and 12 months we measured bone mineral density (BMD) of the lumbar spine, femoral neck and total hip by dual-energy X-ray absorptiometry. All patients had severe osteopenia or osteoporosis at baseline. The risk of femoral neck fracture was calculated at baseline and 12 months (Z-score 2.7). RESULTS: Patients treated with alendronate had a significant increase in BMD at the lumbar spine and femoral neck after 1 year of follow-up, with mean (sd) values of 1.06 (0.26) vs 1.01 (0.21) g/cm(2) at baseline (P < 0.001), and 0.75 (0.07) vs 0.73 (0.07) g/cm(2) (P = 0.03), respectively, while the control group had a significant loss of BMD at the total hip of 0.79 (0.14) vs 0.81 (0.13) g/cm(2) (P = 0.03). BMD was significantly improved at the three locations in patients treated with alendronate compared with the control group, with differences at the lumbar spine, femoral neck and total hip of 0.05 (0.07) vs 0.01 (0.10) (P = 0.001), 0.01 (0.04) vs -0.002 (0.03) (P = 0.04) and 0.01 (0.04) vs -0.01 (0.02) g/cm(2), respectively (P = 0.001). Patients treated with alendronate had a significant decrease in the fracture risk at the femoral neck, by -0.54 (1.29) (P = 0.04) after 1 year of follow-up. CONCLUSIONS: Treatment with once-weekly 70 mg alendronate significantly improved the BMD at the lumbar spine and femoral neck in patients with prostate cancer with severe osteopenia or osteoporosis and on ADT, and significantly decreased the risk of femoral neck fracture. PMID- 19549262 TI - Does the presence of significant risk factors affect perioperative outcomes after robot-assisted radical cystectomy? AB - OBJECTIVE: To evaluate the effect of preoperative risk factors on perioperative outcomes up to 3 months after robot-assisted radical cystectomy (RARC), as RC continues to be associated with a high rate of morbidity and mortality. PATIENTS AND METHODS: From 2005 to 2007, 66 consecutive patients had RARC at Roswell Park Cancer Institute. Patient demographics, preoperative risk factors and complications up to 3 months after RARC were reviewed from a prospective quality assurance database. Patients were stratified into high- and low risk groups based on age, previous abdominal surgery, chronic obstructive pulmonary disease (COPD), body mass index (BMI), Revised Cardiac Risk Index (RCRI) and American Society of Anesthesiologists (ASA) score. RESULTS: Age, previous abdominal surgery, COPD, BMI, RCRI score and ASA score did not significantly influence complications during or up to 3 months following RARC (P > 0.05). Advanced age was associated with a higher RCRI score (P = 0.014) and an increased likelihood of admission to the Intensive Care Unit (P = 0.007). A higher ASA score was associated with an increased overall hospital stay (P = 0.039). Previous abdominal surgery was associated with more frequent unscheduled postoperative clinic visits (P = 0.014). Operative duration did not significantly influence complication rates (P > 0.05). Fifteen of 62 patients (24%) had a major complication, while 15 (24%) had minor complications within 3 months of surgery. The reoperation rate was 11% and the overall mortality rate was 1.6%. CONCLUSIONS: RARC appears to be well tolerated, independent of comorbid risk factors such as age, BMI, RCRI and ASA score. PMID- 19549263 TI - Interferon-induced retinopathy and its risk in patients with diabetes and hypertension undergoing treatment for chronic hepatitis C virus infection. AB - BACKGROUND: Ocular complications are amongst many side-effects of interferon based therapy for hepatitis C virus (HCV) infection. Some suggest that diabetic and hypertensive patients are at increased risk of these complications. AIM: To determine the frequency of ophthalmological complications related to interferon use. METHODS: Retrospective analysis of patients undergoing HCV treatment with pegylated interferon alpha-2a, alpha-2b or consensus interferon plus ribavirin between 2005 and 2007. All patients underwent a baseline eye examination and any visual complaints during treatment prompted a repeat examination. Data recorded included HCV genotype, treatment duration, interferon type, pre-treatment and on treatment visual complaints, known ocular pathology, and retinal findings at baseline and at follow-up. RESULTS: Of 183 patients, 29 (16%) had diabetes and 85 (46%) had hypertension. Seventy-one (38%) received interferon alpha-2a, 100 (55%) alpha-2b, and 12 (7%) consensus interferon. Seven (3.8%) had retinal changes on follow-up and treatment was discontinued in 3 (1.6%). Of seven with ocular changes two had hypertension and one had both hypertension and diabetes. CONCLUSION: The incidence of symptomatic retinopathy in HCV patients undergoing interferon therapy appears low and treatment cessation is rarely needed. Furthermore, patients with hypertension and diabetes may not be at higher risk for interferon-induced retinopathy. PMID- 19549264 TI - Influence of standard treatment on ileal and colonic antimicrobial defensin expression in active Crohn's disease. AB - BACKGROUND: Crohn's Disease (CD), a chronic intestinal inflammation, is currently treated primarily by therapeutics which are directed against inflammatory responses. Recent findings though suggest a central role of the innate immune barrier in the pathophysiology. Important factors providing this barrier are antimicrobial peptides like the alpha- and beta-defensins. Little is known about in vivo effects of common drugs on their expression. AIM: To analyse the influence of corticosteroids, azathioprine and aminosalicylate treatment on ileal and colonic antimicrobial peptides in active CD and also assess the role of inflammation. METHODS: We measured the expression of antimicrobial peptides and pro-inflammatory cytokines in 75 patients with active CD. RESULTS: Ileal and colonic alpha- and beta-defensins as well as LL37 remained unaffected by corticosteroids, azathioprine or aminosalicylate treatment. Additionally, we did not observe a negative coherency between Paneth cell alpha-defensins and any measured cytokines. HBD2 and LL37 unlike HBD1 levels were linked to inflammatory cytokines and increased in highly inflamed samples. CONCLUSIONS: Current oral drug treatment seems to have no major effect on the expression of antimicrobial peptides. In contrast to HBD2 and LL37, ileal levels of HD5 and HD6 and colonic HBD1 level are independent of current inflammation. Innovative drugs should aim to strengthen protective innate immunity. PMID- 19549265 TI - Meta-analysis: clinicopathological and prognostic significance of cyclooxygenase 2 expression on oesophageal squamous cell carcinoma. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) is involved in oesophageal carcinogenesis, but the clinical and prognostic significance of COX-2 expression in oesophageal squamous cell carcinoma (ESCC) remains controversial. AIM: To evaluate the clinicopathological and prognostic role of COX-2 expression in ESCC. METHODS: Studies assessing clinical or prognostic significance of COX-2 expression in ESCC published until December 2008 were selected. A meta-analysis was performed to clarify the impact of COX-2 expression on clinicopathological parameters or overall survival (OS) in ESCC. RESULTS: A total of 19 studies met the inclusion criteria, among which 17 studies were about the clinicopathological significance of COX-2 expression in ESCC, 12 studies were dealing with prognostic role of COX 2 expression in ESCC and 10 studies evaluated both of them. Overexpression of COX 2 was significantly correlated with not only the depth of invasion and TNM stage, with a combined odds ratio (OR) of 0.55 (95%CI: 0.34-0.90, Z = 2.41, P = 0.02) and 0.55 (95%CI: 0.32-0.95, Z = 2.13, P = 0.03) respectively but also the reduced OS with relative risk (RR) 1.42, 95% CI: 1.07-1.90, Z = 2.43, P = 0.02). CONCLUSIONS: COX-2 might play an important role in the progress of ESCC, overexpression of COX-2 correlates with not only the depth of invasion and TNM stage but also the reduced OS. COX-2 might be a potential therapy target for ESCC and work as a prognostic factor for ESCC patients, yet the clinicopathological and prognostic role of COX-2 in ESCC still needs further confirmation by well designed prospective studies. PMID- 19549266 TI - Review article: the current and evolving treatment of colonic diverticular disease. AB - BACKGROUND: Formation of colonic diverticula, via herniation of the colonic wall, is responsible for the development of diverticulosis and consequently diverticular disease. Diverticular disease can be associated with numerous debilitating abdominal and gastrointestinal symptoms (including pain, bloating, nausea, constipation and diarrhoea). AIMS: To review the state of treatment for diverticular disease and its complications, and briefly discuss potential future therapies. METHODS: PubMed and recent conference abstracts were searched for articles describing the treatment of diverticular disease. RESULTS: Many physicians will recommend alterations to lifestyle and increasing fibre consumption. Empirical antibiotics remain the mainstay of therapy for patients with diverticular disease and rifaximin seems to be the best choice. In severe or relapsing disease, surgical intervention is often the only remaining treatment option. Although novel treatment options are yet to become available, the addition of therapies based on mesalazine (mesalamine) and probiotics may enhance treatment efficacy. CONCLUSIONS: Data suggest that diverticular disease may share many of the hallmarks of other, better-characterized inflammatory bowel diseases; however, treatment options for patients with diverticular disease are scarce, revolving around antibiotic treatment and surgery. There is a need for a better understanding of the fundamental mechanisms of diverticular disease to design treatment regimens accordingly. PMID- 19549267 TI - Factors associated with the use of aids to cessation in English smokers. AB - AIMS: To assess factors associated with the use of smoking cessation aids among smokers trying to quit in a country where these aids are widely available and free or cheap to access. DESIGN: Cross-sectional household survey, the 'Smoking Toolkit Study'. SETTING: England. PARTICIPANTS: A total of 3767 respondents who smoked and made at least one serious quit attempt in the past 12 months were interviewed from November 2006 to April 2008. MEASUREMENTS: We analysed differences across socio-demographic and smoking characteristics in the use of nicotine replacement therapy (NRT) over the counter or on prescription, bupropion, varenicline, telephone support and the National Health Service Stop Smoking Service (NHS-SSS) which combines behavioural support with medication. FINDINGS: More than half of smokers trying to quit (51.2%) had used any kind of treatment; 48.4% had used some form of medication but only 6.2% had used the NHS SSS. The use of some form of smoking cessation treatment was higher in female than in male smokers [odds ratio (OR): 1.24, 95% confidence interval (CI): 1.08, 1.43] and increased with age (OR: 1.19, 95% CI: 1.14,1.25) and cigarettes smoked per day (OR = 1.05, 95% CI = 1.04,1.06). There was no association with social grade. Smokers who planned their quit attempt were more likely to have used all types of smoking cessation treatments, except for telephone support. CONCLUSIONS: In England, half of all attempts to quit smoking are aided by some form of pharmacological or behavioural treatment. However, the use of the most effective treatment option (the NHS-SSS) is low, despite it being free of charge. Factors associated with an increased use of aids to cessation were female sex, older age, more cigarettes smoked per day and planning a quit attempt. Research is needed into how to increase utilization rates, particularly among males and younger smokers. PMID- 19549268 TI - Does fibrinogen add to prediction of cardiovascular disease? Results from the Scottish Heart Health Extended Cohort Study. AB - Plasma fibrinogen is an established risk factor for cardiovascular disease (CVD), but it has not been established whether it adds predictive value to risk scores. In the Scottish Heart Health Extended Cohort Study, we measured plasma fibrinogen in 13 060 men and women, aged 30-74 years, initially free of CVD. After follow-up for a median of 19.2 years, 2626 subjects had at least one CVD event. After adjusting for classical CVD risk factors and socio-economic status, the hazard ratios (95% confidence interval) for a one unit (g/l) increase in plasma fibrinogen were 1.09 (1.02, 1.16) for men and 1.10 (1.02, 1.19) for women. Although fibrinogen added significantly to the discrimination of the Framingham risk score for women, it failed to do so for men. Fibrinogen did not add significantly to the ASSIGN risk score. Fibrinogen added between 1.3% and 3.2% to the classification of CVD status by the existing risk scores. We conclude that the added value of fibrinogen to two currently used risk scores is low; hence population screening with fibrinogen for this purpose is unlikely to be clinically useful or cost-effective. PMID- 19549269 TI - The impact of risk stratification by early bone-marrow response in childhood lymphoblastic leukaemia: results from the United Kingdom Medical Research Council trial ALL97 and ALL97/99. AB - The 1997 acute lymphoblastic leukaemia (ALL) trial (ALL97) was a randomised comparison of prednisolone versus dexamethasone and of 6-mercaptopurine versus 6 thioguanine. During the first 2 years of the trial, review of survival data showed the preceding trial, UKALL XI, was no better than its predecessor and that survival for childhood ALL in the UK had not improved in the fashion witnessed by other cooperative treatment groups. The therapy template was therefore altered to an American Children's Cancer Group (CCG) style regimen, including stratification by age, white cell count and early response to therapy by assessment of the bone marrow. This phase of the trial was designated ALL97/99. Comparison of the two phases showed that the event-free survival (EFS) for both ALL97 and ALL97/99 was better than previous UKALL trials, as was overall survival (OS) for ALL97/99. Both EFS and OS were significantly better in ALL97/99 than in ALL97 (at five years, 80.0% vs. 74.0%, P = 0.002; and 88.0% vs. 83.5%, P = 0.005, respectively). Isolated central nervous system (CNS) relapse for patients in ALL97/99 was half that in ALL97 (3.0% vs. 4.9%), P = 0.03) and the overall CNS relapse rate was halved in ALL97/99 (4.4% vs. 9.6%, P < 0.00005). There were no significant differences for non-CNS relapse, induction deaths or deaths in remission between the two phases of the trial. PMID- 19549270 TI - Aminophospholipid translocase and phospholipid scramblase activities in sickle erythrocyte subpopulations. AB - Phosphatidylserine (PS) externalization may contribute to Sickle Cell Disease (SCD) characteristics including thrombogenesis, endothelial adhesion and shortened red blood cell (RBC) lifespan. Aminophospholipid translocase (APLT) returns externalized PS to the inner membrane, and phospholipid scramblase (PLSCR) equilibrates phospholipids (PL) across the membrane. APLT inhibition and PLSCR activation appear to be important for PS externalization. We examined relationships between APLT, PLSCR and external PS in mature sickle RBC and reticulocytes. Normally-hydrated sickle RBC without external PS had active APLT and inactive PLSCR. PS-exposing sickle RBC had inhibited APLT and active PLSCR. Sickle reticulocytes had active APLT and active PLSCR independent of external PS. Sickle RBC dehydrated in vivo had the highest proportion of PS-exposing RBC and markedly inhibited APLT. Normal and sickle RBC dehydrated in vitro had moderately decreased APLT. Rehydration resulted in significant recovery of APLT in RBC previously dehydrated in vitro, but not in sickle RBC dehydrated in vivo. These findings indicate that (i) PS externalization in mature sickle RBC depends on the balance between APLT and PLSCR activities, (ii) PS externalization in sickle reticulocytes depends primarily on PLSCR activation and (iii) APLT inhibition in sickle RBC dehydrated in vivo is due to dehydration itself and other factors. PMID- 19549273 TI - Right coronary fistula and aneurysm draining to the right atrium. AB - In a 3-year-old boy, a continuous heart murmur was heard. The echocardiogram showed a dilated right coronary artery suggesting the existence of a coronary fistula. A more detailed echocardiogram when the patient was sedated revealed a fistula leading to a large aneurysm and further to the right atrium. The accidental dissection and thrombosis during the interventional heart catheterization resulted in a closure of the fistula. A continuous heart murmur and a dilated coronary artery are the hallmarks of coronary fistula. CONCLUSION: Anatomic details of coronary fistula might be possible to see in an echocardiogram. Interventional heart catheterization is usually an adequate treatment option. PMID- 19549271 TI - Outcome, timing and adverse events in therapeutic hypothermia after out-of hospital cardiac arrest. AB - BACKGROUND: Therapeutic hypothermia (TH) after cardiac arrest protects from neurological sequels and death and is recommended in guidelines. The Hypothermia Registry was founded to the monitor outcome, performance and complications of TH. METHODS: Data on out-of-hospital cardiac arrest (OHCA) patients admitted to intensive care for TH were registered. Hospital survival and long-term outcome (6 12 months) were documented using the Cerebral Performance Category (CPC) scale, CPC 1-2 representing a good outcome and 3-5 a bad outcome. RESULTS: From October 2004 to October 2008, 986 TH-treated OHCA patients of all causes were included in the registry. Long-term outcome was reported in 975 patients. The median time from arrest to initiation of TH was 90 min (interquartile range, 60-165 min) and time to achieving the target temperature (< or =34 degrees C) was 260 min (178 400 min). Half of the patients underwent coronary angiography and one-third underwent percutaneous coronary intervention (PCI). Higher age, longer time to return of spontaneous circulation, lower Glasgow Coma Scale at admission, unwitnessed arrest and initial rhythm asystole were all predictors of bad outcome, whereas time to initiation of TH and time to reach the goal temperature had no significant association. Bleeding requiring transfusion occurred in 4% of patients, with a significantly higher risk if angiography/PCI was performed (2.8% vs. 6.2%P=0.02). CONCLUSIONS: Half of the patients survived, with >90% having a good neurological function at long-term follow-up. Factors related to the timing of TH had no apparent association to outcome. The incidence of adverse events was acceptable but the risk of bleeding was increased if angiography/PCI was performed. PMID- 19549274 TI - Chronic mucocutaneous candidiasis may cause elevated gliadin antibodies. AB - We present a 4-year-old boy admitted to the hospital due to the typical symptoms of celiac disease with severe dystrophy, anaemia and elevated gliadin IgG antibodies. Upper endoscopy ruled out celiac disease but showed severe Candida esophagitis. Due to an impaired T-cell function especially following Candida antigen stimulation in vitro, plus recurrent Candida infections of the skin, the diagnosis of chronic mucocutaneous candidasis (CMC) was made. Under the treatment with fluconazol, trimethoprim/sulfmethoxazole and IVIG, the child improved impressively. Gliadin antibodies declined steadily. CONCLUSION: The common symptoms growth retardation, anaemia and elevated gliadin antibodies are suggestive for celiac disease but very unspecific. The rare immunodeficiency CMC may cause elevated gliadin antibodies. PMID- 19549275 TI - Self-concept and mental health status of 'stay-at-home' children in rural China. AB - AIM: To describe the self-concept and mental health status of 'stay-at-home' children and to explore the differences between stay-at-home children and non stay-at-home children. METHODS: A cross-sectional survey was conducted in Changfeng County to collect information on self-concept and mental health status. Children were classified as 'stay-at-home' or 'non-stay-at-home' for data analysis. RESULTS: Stay-at-home children accounted for 55.1% of children. The two groups of children differed significantly on the total scores of self-concept (stay-at-home, 52.48 +/- 14.29; non-stay-at-home, 55.24 +/- 15.10). The mental health status of stay-at-home children was poor, with significant difference between them (stay-at-home, 41.17 +/- 12.25; non-stay-at-home, 40.14 +/- 13.11). Using multivariate linear regression analysis, we found that the total P-H score, gender, low family economic status, stay-at-home status and being cared for by an uncle/aunt or an older sibling were independent variables for mental health of the children. CONCLUSION: This study suggests that stay-at-home children have a greater risk of mental health problems than their counterparts in rural Anhui province, China. In addition, this study provides useful baseline information on childhood mental health and has identified important risk factors that would be important in planning strategies for prevention of mental health problems for stay-at-home children. PMID- 19549276 TI - Thrombin generation and heparin-induced thrombocytopenia. AB - BACKGROUND: Heparin-induced thrombocytopenia (HIT) is a severe complication of heparin therapy. IgG antibodies targeting the platelet factor 4-heparin complex activate platelets and generate microparticles with procoagulant activity. OBJECTIVES: To determine whether the thrombin generation assay is capable of detecting procoagulant activity induced by patient platelet-poor plasma (PPP) in donor platelet-rich plasma (PRP). PATIENTS AND METHODS: We explored two groups of patients; group 1 (n = 23): patients with a positive clinical and biological diagnosis of HIT; group 2 (n = 25): patients with a negative clinical and biological diagnosis of HIT. Mixtures of donor PRP and patient PPP (1:1) were incubated either with unfractionated heparin 0.2 U mL(-1) or with physiological saline. Thrombin generation was assessed by calibrated thrombinography. The effect of heparin on the mixtures was evaluated according to the ratio of the values with and without heparin (wH/woH) of the five thrombogram parameters. RESULTS: With low heparin concentrations, plasma of group 1 activates donor platelets and generates procoagulant activity. A set of three ratios outside the cut-off values corresponds to the 'HIT thrombogram profile', characterized by a highly specific aspect of the thrombogram wH in relation to the thrombogram woH. None of the group 2 patients presented a HIT thrombogram profile. The results of thrombinography correlate well with the results of the platelet aggregation test. CONCLUSION: Our studies illustrate the central paradox of HIT, namely enhancement of thrombin generation in the presence of heparin. The HIT thrombogram profile as it is defined in this study can detect the procoagulant activity of HIT IgG antibodies. PMID- 19549277 TI - Natural history of TFR2-related hereditary hemochromatosis in a 47-yr-old Italian patient. PMID- 19549278 TI - The impact of cytogenetic abnormalities on the prognosis of primary myelofibrosis: a prospective survey of 202 cases in Japan. AB - Cytogenetic abnormalities were often observed in primary myelofibrosis patients. The presence of specific cytogenetic abnormalities, such as sole abnormalities of chromosome 13q-, 20q-, or -7/7q-, is reported to have the influence on the prognosis of primary myelofibrosis. We analyzed the data from the prospective survey of Japanese primary myelofibrosis patients which was conducted from 1999 to clarify the impact of cytogenetic abnormalities on the prognosis of primary myelofibrosis. A total of 202 primary myelofibrosis patients had the cytogenetic and the prognostic data. Eighty (40%) out of 202 cases had cytogenetic abnormalities, and an association was evident for platelet counts. Although the presence of an abnormal karyotype did not affect the prognosis, primary myelofibrosis patients with cytogenetic abnormalities other than 13q- and 20q- showed an inferior prognosis compared to patients with a normal karyotype or sole 13q- or 20q- abnormalities. Patients with an unfavorable cytogenetic profile (abnormal cytogenetics other than 13q- or 20q-) also had a greater tendency to transform to leukemia than patients with a favorable cytogenetic profile (normal cytogenetics, sole abnormalities of either chromosome 13q-, or 20q-). Abnormal cytogenetics other than 13q- or 20q- in primary myelofibrosis patients has the poor prognostic effect for both survival and the risk of leukemic transformation. PMID- 19549280 TI - Bioactive TGF-beta can associate with lipoproteins and is enriched in those containing apolipoprotein E3. AB - Transforming growth factor-beta1 (TGF-beta1) has central functions in development, tissue maintenance, and repair and has been implicated in major diseases. We discovered that TGF-beta1 contains several amphipathic helices and hydrophobic domains similar to apolipoprotein E (apoE), a protein involved in lipoprotein metabolism. Indeed, TGF-beta1 associates with lipoproteins isolated from human plasma, cultured liver cells, or astrocytes, and its bioactivity was highest in high-density lipoprotein preparations. Importantly, lipoproteins containing the apoE3 isoform had higher TGF-beta levels and bioactivity than those containing apoE4, a major genetic risk factor for atherosclerosis and Alzheimer's disease. Because TGF-beta1 can be protective in these diseases an association with apoE3 may be beneficial. Association of TGF-beta with different types of lipoproteins may facilitate its diffusion, regulate signaling, and offer additional specificity for this important growth factor. PMID- 19549281 TI - Inhibition of tau fibrillization by oleocanthal via reaction with the amino groups of tau. AB - Tau is a microtubule-associated protein that promotes microtubule assembly and stability. In Alzheimer's disease and related tauopathies, tau fibrillizes and aggregates into neurofibrillary tangles. Recently, oleocanthal isolated from extra virgin olive oil was found to display non-steroidal anti-inflammatory activity similar to ibuprofen. As our unpublished data indicates an inhibitory effect of oleocanthal on amyloid beta peptide fibrillization, we reasoned that it might inhibit tau fibrillization as well. Herein, we demonstrate that oleocanthal abrogates fibrillization of tau by locking tau into the naturally unfolded state. Using PHF6 consisting of the amino acid residues VQIVYK, a hexapeptide within the third repeat of tau that is essential for fibrillization, we show that oleocanthal forms an adduct with the lysine via initial Schiff base formation. Structure and function studies demonstrate that the two aldehyde groups of oleocanthal are required for the inhibitory activity. These two aldehyde groups show certain specificity when titrated with free lysine and oleocanthal does not significantly affect the normal function of tau. These findings provide a potential scheme for the development of novel therapies for neurodegenerative tauopathies. PMID- 19549283 TI - An epidemiological study to identify the risk factors with two different types of controls in high-grade cervical lesions including invasive cancer. AB - A multidisciplinary study on pre-cancerous and early cancerous lesions of uterine cervix was carried out at our Institute from which the subjects (cases and one group of control) for the present study were selected with the objective to identify the possible risk factors related to high-grade cervical lesions including invasive cancer through an epidemiological study by selecting two different types of controls and to assess the feasibility whether the cancers of other organs could be taken as controls at the same time studying the risk factors associated with cervical cancer. One group of control was women with negative Pap smear and second group of control was the women with breast cancer but negative Pap smear. A total of 100 biopsy-proven cases of high-grade cervical intraepithelial lesions and Invasive cancer were recruited. The results of the study show that some of the risk factors associated with the cervical cancer get exaggerated when breast cancer cases were used as controls because risk factors of both cancers are opposite to each other. So it is concluded that in order to remove any bias, normal hospital controls or controls selected from multiple cancers should be taken to study the risk factors involved in cervical carcinogenesis. PMID- 19549279 TI - Circadian regulation of ion channels and their functions. AB - Ion channels are the gatekeepers to neuronal excitability. Retinal neurons of vertebrates and invertebrates, neurons of the suprachiasmatic nucleus (SCN) of vertebrates, and pinealocytes of non-mammalian vertebrates display daily rhythms in their activities. The interlocking transcription-translation feedback loops with specific post-translational modulations within individual cells form the molecular clock, the basic mechanism that maintains the autonomic approximately 24-h rhythm. The molecular clock regulates downstream output signaling pathways that further modulate activities of various ion channels. Ultimately, it is the circadian regulation of ion channel properties that govern excitability and behavior output of these neurons. In this review, we focus on the recent development of research in circadian neurobiology mainly from 1980 forward. We will emphasize the circadian regulation of various ion channels, including cGMP gated cation channels, various voltage-gated calcium and potassium channels, Na(+)/K(+)-ATPase, and a long-opening cation channel. The cellular mechanisms underlying the circadian regulation of these ion channels and their functions in various tissues and organisms will also be discussed. Despite the magnitude of chronobiological studies in recent years, the circadian regulation of ion channels still remains largely unexplored. Through more investigation and understanding of the circadian regulation of ion channels, the future development of therapeutic strategies for the treatment of sleep disorders, cardiovascular diseases, and other illnesses linked to circadian misalignment will benefit. PMID- 19549282 TI - The HDAC inhibitor, sodium butyrate, stimulates neurogenesis in the ischemic brain. AB - In the healthy adult brain, neurogenesis normally occurs in the subventricular zone (SVZ) and hippocampal dentate gyrus (DG). Cerebral ischemia enhances neurogenesis in neurogenic and non-neurogenic regions of the ischemic brain of adult rodents. This study demonstrated that post-insult treatment with a histone deacetylase inhibitor, sodium butyrate (SB), stimulated the incorporation of bromo-2'-deoxyuridine (BrdU) in the SVZ, DG, striatum, and frontal cortex in the ischemic brain of rats subjected to permanent cerebral ischemia. SB treatment also increased the number of cells expressing polysialic acid-neural cell adhesion molecule, nestin, glial fibrillary acidic protein, phospho-cAMP response element-binding protein (CREB), and brain-derived neurotrophic factor (BDNF) in various brain regions after cerebral ischemia. Furthermore, extensive co localization of BrdU and polysialic acid-neural cell adhesion molecule was observed in multiple regions after ischemia, and SB treatment up-regulated protein levels of BDNF, phospho-CREB, and glial fibrillary acidic protein. Intraventricular injection of K252a, a tyrosine kinase B receptor antagonist, markedly reduced SB-induced cell proliferation detected by BrdU and Ki67 in the ipsilateral SVZ, DG, and other brain regions, blocked SB-induced nestin expression and CREB activation, and attenuated the long-lasting behavioral benefits of SB. Together, these results suggest that histone deacetylase inhibitor-induced cell proliferation, migration and differentiation require BDNF tyrosine kinase B signaling and may contribute to long-term beneficial effects of SB after ischemic injury. PMID- 19549284 TI - Positron emission tomography/computed tomography introduction in the clinical management of patients with suspected recurrence of ovarian cancer: a cost effectiveness analysis. AB - Aim of this study was to evaluate the economic impact of the introduction of positron emission tomography/computed tomography (PET/CT) in the early detection of recurrent ovarian cancer through a cost-effectiveness analysis of different diagnostic strategies. Thirty-two consecutive patients with suspected ovarian cancer recurrence, studied by both contrast enhanced abdominal CT and PET/CT, were retrospectively included in the study. Three different diagnostic strategies were evaluated and compared: (1) CT only or baseline strategy; (2) PET/CT for negative CT or strategy A; (3) PET/CT for All or strategy B. For each one, expected costs, avoided surgery and incremental cost-effectiveness ratio (ICER) were calculated to identify the most cost-effective strategy. The number of positive patients increased from baseline strategy (20/32) to strategy A and B (30/32 and 29/32 respectively). Positron emission tomography/computed tomography reoriented physician choice in 31% and 62% of patients (strategies A and B respectively). Strategy A is dominated by strategy B, which is more expensive (2909 euro vs. 2958 euro), but also more effective (3 cases of surgery avoided) and presents an ICER of 226.77 euro per surgery avoided (range: 49.50-433.00 euro). Positron emission tomography/computed tomography introduction in this population is cost-effective and allowed to redirect the clinical management of patients towards more appropriate therapeutic choices. PMID- 19549285 TI - Supraclavicular lymph node metastases of unknown origin: HPV-typing identifies the primary tumour. AB - Cancers of unknown primary origin (CUP) account for 0.5-10% of all malignancies. CUP patients with metastases have a median survival of approximately 6 months, despite therapy. Identification of the primary tumour site may offer the opportunity of a specific and more efficient treatment. The case of a 45-year-old woman with supraclavicular lymph node metastases of a squamous cell CUP is reported. A staging laparoscopy with multiple biopsies and a loop diathermy excision of the cervix were performed. Human papillomavirus (HPV)-testing in the tissues revealed the tumour cells as metastases of an occult cervical cancer. Primary platin-based chemotherapy combined with paclitaxel leads to a complete apparative remission. Twelve months later, staging positron emission tomography with 2-[18F]fluoro-2-deoxy-D-glucose in combination with computed tomography identified an isolated left renal lymph node metastasis. The patient received targeted radiation therapy, combined with cisplatin. To date, 19 months after diagnosis, she is doing well without any evidence of disease. The presented case report addresses the difficulties involving the identification of CUP. HPV-DNA is found in over 95% of cervical cancers. As the presented case illustrates, testing for this virus DNA in human tissues can be a useful diagnostic tool in patients with CUP where cervical cancer is the possible primary tumour. PMID- 19549286 TI - Assessing the unmet supportive care needs of newly diagnosed patients with cancer. AB - Adequate monitoring in cancer control needs to include measures of psychosocial outcomes so as to take account of the totality of the felt cancer experience. There is a need to know whether the experience of cancer is changing, as well as a need for tools to identify where supportive care interventions or services could be targeted to good effect. The aim of this study was to investigate the utility of using a telephone-administered survey to identify the unmet needs of cancer patients. Participants were identified from a statewide population-based cancer registry following an episode of hospitalized care in Victoria (Australia). Of the 506 eligible patients, 236 completed a telephone adaptation of the Supportive Care Needs Survey-Short Form (SCNS-SF31). Sampling from the cancer registry was shown to provide timely contact with patients. Results from the survey indicated that perceived needs for this newly diagnosed group of cancer patients were mostly in the area of information provision. Results also showed that some socio-demographic and disease-specific variables affected the level of perceived unmet needs. Overall, this study indicated that registry-based sampling was practical and the telephone adaptation of the SCNS-SF31 provided a reliable method to explore the unmet needs of newly diagnosed patients with cancer. PMID- 19549287 TI - Clinical trial: prophylactic intravenous alanyl-glutamine reduces the severity of gastrointestinal toxicity induced by chemotherapy--a randomized crossover study. AB - BACKGROUND: Glutamine has been shown in numerous studies to reduce intestinal permeability which can be increased by chemotherapy. However, there have been few reports that conduct on its clinical effect on gastrointestinal toxicity. AIM: To examine whether prophylactic intravenous alanyl-glutamine dipeptide can ameliorate clinical manifestations of gastrointestinal toxicity induced by chemotherapy. METHODS: Forty-four patients with gastric or colorectal cancer developing WHO side-effect grading system of grade 2 or higher were randomized to either control group (n = 22) or Gln group (n = 22) during next cycle of chemotherapy. Patients were crossed over to the alternate treatment during chemotherapy cycle 2. In the control group, the patients received the same chemotherapy regimens as screening cycle and in the Gln group, the patients received chemotherapy and alanyl-glutamine. Prophylactic intravenous 20 g of alanyl-glutamine dipeptide was given for 5 days. RESULTS: Compared with the control group, the plasma glutamine level in the Gln group was significantly higher and the plasma endotoxin level was significantly lower. The scores of nausea/vomiting and diarrhoea decreased significantly. CONCLUSION: Prophylactic intravenous alanyl-glutamine is effective in preventing intestinal permeability disruption induced by chemotherapy and clinical manifestations of gastrointestinal toxicity. PMID- 19549288 TI - The use of exclusive enteral nutrition for induction of remission in children with Crohn's disease demonstrates that disease phenotype does not influence clinical remission. AB - BACKGROUND: Exclusive enteral nutrition (EEN) achieves variable remission rates in patients with Crohn's disease (CD). AIM: To describe our experience of treating CD with an 8-week course of primary EEN and to study factors affecting treatment outcome. METHODS: All CD patients treated with EEN in our centre between 2004 and 2007 were included in the study. Remission was determined by a combination of clinical parameters. Disease phenotype was assigned using published classifications. Inflammatory markers and anthropometry (Z-scores) were calculated before and after treatment. RESULTS: A total of 114 children were treated (four were excluded). Median age at diagnosis was 11.6 years. Fifty-seven (51.8%) were fed orally whilst 53 (48.2%) were fed by tube. Eighty-eight (80%) achieved remission with consequent reductions in erythrocyte sedimentation rate and C-reactive protein (P < 0.001). Patients in remission had comparative improvements in weight (-1.04 cf. -0.40) and BMI Z-scores (-0.98 cf. -0.03) by the end of treatment (P < 0.001). Individuals with isolated terminal ileal disease (n = 4) had lower remission rates than other locations (P = 0.02). No other significant differences in remission rates for any other disease locations were found. CONCLUSIONS: Exclusive enteral nutrition induces clinical remission, normalization of inflammatory markers and improves weight/BMI Z-scores in most patients. This study demonstrates that disease phenotype should not influence clinicians when commencing patients on EEN. PMID- 19549289 TI - The genetic architecture of reproductive isolation in Louisiana irises: hybrid fitness in nature. AB - Negative epistasis in hybrid genomes commonly results in postzygotic isolation between divergent lineages. However, some genomic regions may be selectively neutral or adaptive in hybrids and thus may potentially cross species barriers. We examined postzygotic isolation between ecologically similar species of Louisiana Iris: Iris brevicaulis and I. fulva to determine the potential for adaptive introgression in nature. Line-cross analyses allowed us a general overview of the gene action responsible for fitness-related traits. We then used a QTL mapping approach to detect genomic regions responsible for variation in these traits. Although hybrid classes suffered reduced fitness for many traits, hybrid means were equivalent to at least one of the parental species in overall estimates of maternal and paternal fitness during the two years of the field study. The genetic architecture underlying the fitness-related traits varied across field site and year of the study, thus emphasizing the importance of the environment in determining the degree of postzygotic isolation and potential for introgression across natural hybrid zones. PMID- 19549290 TI - Masitinib in the treatment of active rheumatoid arthritis: results of a multicentre, open-label, dose-ranging, phase 2a study. AB - INTRODUCTION: Since current treatment options for patients suffering from active rheumatoid arthritis (RA) remain inadequate, especially for those unresponsive to disease-modifying antirheumatic drugs (DMARDs), new and improved medication is needed. This study evaluates the safety and efficacy of masitinib (AB1010), a potent and selective protein tyrosine kinase inhibitor of c-KIT, in the monotherapy treatment of DMARD-refractory RA. METHODS: This was a multicentre, uncontrolled, open-label, randomised, dose-ranging, phase 2a trial. Masitinib was administered orally to 43 patients who had inadequate response to DMARDs, at initial randomised dosing levels of 3 and 6 mg/kg per day over a 12-week period. Dose adjustment was permitted based upon tolerability and response criteria. Efficacy was assessed via American College of Rheumatology 20%/50%/70% improvement criteria (ACR20/50/70) responses, disease activity score using 28 joint counts (DAS28), index of improvement in RA (ACRn) and C-reactive protein (CRP) improvement, relative to baseline at week 12. RESULTS: Improvement was observed in all efficacy endpoints, including ACR20/50/70 scores of 54%, 26% and 8%, respectively, and a reduction in CRP level by greater than 50% for approximately half the population. This improvement was sustainable throughout an extension phase (> 84 weeks) and was also independent of initial DMARD resistance (anti-tumour necrosis factor-alpha and/or methotrexate). A relatively high patient withdrawal rate (37%) required the use of last observation carried forward (LOCF) data imputation. Incidence of adverse events was high (95%), although the majority were of mild or moderate severity with a considerable decline in frequency observed after 12 weeks of treatment. Two nonfatal serious adverse events were reported. Dose-response analyses tentatively indicate that an initial dosing level of 6.0 mg/kg per day administered orally in two daily intakes is the most appropriate, based upon potency and tolerability trends. CONCLUSIONS: Treatment with masitinib improved DMARD-refractory active RA. Following an initial high incidence of mostly mild to moderate side effects during the first 12 weeks of treatment, masitinib appears to be generally well tolerated. This, together with evidence of a sustainable efficacy response, suggests that masitinib is suitable for long-term treatment regimens. Since this was the first study of masitinib in a nononcologic pathology, the relatively high patient withdrawal rate observed can be partly attributed to a highly cautious response to adverse events. There is sufficient compelling evidence to warrant further placebo-controlled investigation. TRIAL REGISTRATION: ClinicalTrials.gov NCT00831922. PMID- 19549291 TI - A comprehensive resource for integrating and displaying protein post translational modifications. AB - BACKGROUND: Protein Post-Translational Modification (PTM) plays an essential role in cellular control mechanisms that adjust protein physical and chemical properties, folding, conformation, stability and activity, thus also altering protein function. FINDINGS: dbPTM (version 1.0), which was developed previously, aimed on a comprehensive collection of protein post-translational modifications. In this update version (dbPTM2.0), we developed a PTM database towards an expert system of protein post-translational modifications. The database comprehensively collects experimental and predictive protein PTM sites. In addition, dbPTM2.0 was extended to a knowledge base comprising the modified sites, solvent accessibility of substrate, protein secondary and tertiary structures, protein domains, protein intrinsic disorder region, and protein variations. Moreover, this work compiles a benchmark to construct evaluation datasets for computational study to identifying PTM sites, such as phosphorylated sites, glycosylated sites, acetylated sites and methylated sites. CONCLUSION: The current release not only provides the sequence based information, but also annotates the structure-based information for protein post-translational modification. The interface is also designed to facilitate the access to the resource. This effective database is now freely accessible at http://dbPTM.mbc.nctu.edu.tw/. PMID- 19549292 TI - Validation of a primer optimisation matrix to improve the performance of reverse transcription - quantitative real-time PCR assays. AB - BACKGROUND: The development of reverse transcription - quantitative real-time PCR (RT-qPCR) platforms that can simultaneously measure the expression of multiple genes is dependent on robust assays that function under identical thermal cycling conditions. The use of a primer optimisation matrix to improve the performance of RT-qPCR assays is often recommended in technical bulletins and manuals. Despite this recommendation, a comprehensive introduction to and evaluation of this approach has been absent from the literature. Therefore, we investigated the impact of varying the primer concentration, leaving all the other reaction conditions unchanged, on a large number of RT-qPCR assays which in this case were designed to be monitored using hydrolysis probes from the Universal Probe Library (UPL) library. FINDINGS: Optimal RT-qPCR conditions were determined for 60 newly designed assays. The calculated Cq (Quantification Cycle) difference, non specific amplification, and primer dimer formation for a given assay was often dependent on primer concentration. The chosen conditions were further optimised by testing two different probe concentrations. Varying the primer concentrations had a greater effect on the performance of a RT-qPCR assay than varying the probe concentrations. CONCLUSION: Primer optimisation is important for improving the performance of RT-qPCR assays monitored by UPL probes. This approach would also be beneficial to the performance of other RT-qPCR assays such as those using other types of probes or fluorescent intercalating dyes. PMID- 19549293 TI - Homicide and geographic access to gun dealers in the United States. AB - BACKGROUND: Firearms are the most commonly used weapon to commit homicide in the U.S. Virtually all firearms enter the public marketplace through a federal firearms licensee (FFL): a store or individual licensed by the federal government to sell firearms. Whether FFLs contribute to gun-related homicide in areas where they are located, in which case FFLs may be a homicide risk factor that can be modified, is not known. METHODS: Annual county-level data (1993-1999) on gun homicide rates and rates of FFLs per capita were analyzed using negative binomial regression controlling for socio-demographic characteristics. Models were run to evaluate whether the relation between rates of FFLs and rates of gun homicide varied over the study period and across counties according to their level of urbanism (defined by four groupings, as below). Also, rates of FFLs were compared against FS/S - which is the proportion of suicides committed by firearm and is thought to be a good proxy for firearm availability in a region - to help evaluate how well the FFL variable is serving as a way to proxy firearm availability in each of the county types of interest. RESULTS: In major cities, gun homicide rates were higher where FFLs were more prevalent (rate ratio [RR] = 1.70, 95% CI 1.03-2.81). This association increased (p < 0.01) from 1993 (RR = 1.69) to 1999 (RR = 12.72), due likely to federal reforms that eliminated low volume dealers, making FFL prevalence a more accurate exposure measure over time. No association was found in small towns. In other cities and in suburbs, gun homicide rates were significantly lower where FFLs were more prevalent, with associations that did not change over the years of the study period. FFL prevalence was correlated strongly (positively) with FS/S in major cities only, suggesting that the findings for how FFL prevalence relates to gun homicide may be valid for the findings pertaining to major cities but not to counties of other types. CONCLUSION: Modification of FFLs through federal, state, and local regulation may be a feasible intervention to reduce gun homicide in major cities. PMID- 19549294 TI - Effectiveness of physical training for self-employed persons with musculoskeletal disorders: a randomized controlled trial. AB - BACKGROUND: Despite the fact that the population of self-employed persons is still growing and at risk for long term disability due to a number of risk factors, there is still a lack of information on the effectiveness of interventions for this specific group. METHODS: To determine the effectiveness of physical training without a cognitive behavioral component and workplace specific exercises (PT) and physical training with a cognitive behavioral component and workplace specific exercises (PTCBWE), we conducted a pragmatic Randomized Controlled Trial, stratified into two groups. Self-employed persons with a new work disability claim because of musculoskeletal disorders were randomized to PT (n = 53) or PTCBWE (n = 76), or to a corresponding usual care group (n = 50 and n = 75 respectively). Both types of training consisted of cardiovascular training, strengthening, relaxation and posture exercises and took place two or three times a week, for 1-1.5 hours, during three months, also if someone had already returned to work full-time. The primary outcome measure was claim duration (in days) during 12 months follow-up. Pain severity and functional status were secondary outcome measures. All data were assessed at baseline and at 6 and 12 months follow-up. The data with regard to claim duration were analyzed by survival analysis and Cox regression analysis. Secondary outcome measures were analyzed by means of linear regression analysis. RESULTS: After 12 months of follow-up there was no difference in claim duration between PT and usual care (Hazard Ratio 0.7; 95%CI, 0.4-1.1; p = 0.12) or PTCBWE and usual care (Hazard Ratio 0.9; 95%CI, 0.6-1.4; p = 0.72). Both types of physical training and usual care improved in pain and functional status over time, but there was only a statistically significant difference in favor of PT on pain improvement at 6 months. CONCLUSION: In this study, physical training with and without a cognitive behavioral component and workplace specific exercises for self-employed persons with musculoskeletal disorders was not shown to be effective on claim duration, pain severity and functional status at 12 months follow-up. TRIAL REGISTRATION: Current Controlled Trials ISRCTN67766245. PMID- 19549295 TI - Role of serotonergic neurons in the Drosophila larval response to light. AB - BACKGROUND: Drosophila larval locomotion consists of forward peristalsis interrupted by episodes of pausing, turning and exploratory behavior (head swinging). This behavior can be regulated by visual input as seen by light induced increase in pausing, head swinging and direction change as well as reduction of linear speed that characterizes the larval photophobic response. During 3rd instar stage, Drosophila larvae gradually cease to be repelled by light and are photoneutral by the time they wander in search for a place to undergo metamorphosis. Thus, Drosophila larval photobehavior can be used to study control of locomotion. RESULTS: We used targeted neuronal silencing to assess the role of candidate neurons in the regulation of larval photobehavior. Inactivation of DOPA decarboxylase (Ddc) neurons increases the response to light throughout larval development, including during the later stages of the 3rd instar characterized by photoneutral response. Increased response to light is characterized by increase in light-induced direction change and associated pause, and reduction of linear movement. Amongst Ddc neurons, suppression of the activity of corazonergic and serotonergic but not dopaminergic neurons increases the photophobic response observed during 3rd instar stage. Silencing of serotonergic neurons does not disrupt larval locomotion or the response to mechanical stimuli. Reduced serotonin (5-hydroxytryptamine, 5-HT) signaling within serotonergic neurons recapitulates the results obtained with targeted neuronal silencing. Ablation of serotonergic cells in the ventral nerve cord (VNC) does not affect the larval response to light. Similarly, disruption of serotonergic projections that contact the photoreceptor termini in the brain hemispheres does not impact the larval response to light. Finally, pan-neural over-expression of 5-HT1A Dro receptors, but not of any other 5-HT receptor subtype, causes a significant decrease in the response to light of 3rd instar larvae. CONCLUSION: Our data demonstrate that activity of serotonergic and corazonergic neurons contribute to the control of larval locomotion by light. We conclude that this control is carried out by 5-HT neurons located in the brain hemispheres, but does not appear to occur at the photoreceptor level and may be mediated by 5-HT1A Dro receptors. These findings provide new insights into the function of 5-HT neurons in Drosophila larval behavior as well as into the mechanisms underlying regulation of larval response to light. PMID- 19549296 TI - A new surgical ventricular restoration technique to reset residual myocardium's fiber orientation: the "KISS" procedure. AB - BACKGROUND: The history of surgical reconstruction of the left ventricle after an anterior myocardial infarction shows an evolution of techniques which tend to a more and more physiologic restoration of ventricular shape and volume, with increasing attention to the orientation of myocardial fibers. METHODS: We set a new surgical procedure for endoventricular patch reconstruction technique with the aim to rebuild a physiologic shape and volume of the left ventricle caring about realignment of myocardial fibers orientation. Peculiarities of this reconstruction are the shape of the patch (reduction of minor axis compared with currently used oval-shaped patch) and the asymmetrical way of suturing it inside the ventricle. RESULTS: We present a detailed description of operative steps of this procedure, and we add some relevant surgical hints to clarify its peculiarities. Most of the patients operated on with this technique showed the original renewal of apical rotation and left ventricular torsion as specific index of the restoration of physiologic fiber orientation: we report an exemplary case of at-sight recovery of apical rotation in the operating room. CONCLUSION: This technique can represent a reproducible new way to realign myocardial fibers in a near-normal setting, improving the physiological restoration of ischemically injured left ventricle. It could be also the basis to reconsider surgical treatment for heart failure. PMID- 19549297 TI - Geostatistical evaluation of integrated marsh management impact on mosquito vectors using before-after-control-impact (BACI) design. AB - BACKGROUND: In many parts of the world, salt marshes play a key ecological role as the interface between the marine and the terrestrial environments. Salt marshes are also exceedingly important for public health as larval habitat for mosquitoes that are vectors of disease and significant biting pests. Although grid ditching and pesticides have been effective in salt marsh mosquito control, marsh degradation and other environmental considerations compel a different approach. Targeted habitat modification and biological control methods known as Open Marsh Water Management (OMWM) had been proposed as a viable alternative to marsh-wide physical alterations and chemical control. However, traditional larval sampling techniques may not adequately assess the impacts of marsh management on mosquito larvae. To assess the effectiveness of integrated OMWM and marsh restoration techniques for mosquito control, we analyzed the results of a 5-year OMWM/marsh restoration project to determine changes in mosquito larval production using GIS and geostatistical methods. METHODS: The following parameters were evaluated using "Before-After-Control-Impact" (BACI) design: frequency and geographic extent of larval production, intensity of larval production, changes in larval habitat, and number of larvicide applications. The analyses were performed using Moran's I, Getis-Ord, and Spatial Scan statistics on aggregated before and after data as well as data collected over time. This allowed comparison of control and treatment areas to identify changes attributable to the OMWM/marsh restoration modifications. RESULTS: The frequency of finding mosquito larvae in the treatment areas was reduced by 70% resulting in a loss of spatial larval clusters compared to those found in the control areas. This effect was observed directly following OMWM treatment and remained significant throughout the study period. The greatly reduced frequency of finding larvae in the treatment areas led to a significant decrease (approximately 44%) in the number of times when the larviciding threshold was reached. This reduction, in turn, resulted in a significant decrease (approximately 74%) in the number of larvicide applications in the treatment areas post-project. The remaining larval habitat in the treatment areas had a different geographic distribution and was largely confined to the restored marsh surface (i.e. filled-in mosquito ditches); however only approximately 21% of the restored marsh surface supported mosquito production. CONCLUSION: The geostatistical analysis showed that OMWM demonstrated considerable potential for effective mosquito control and compatibility with other natural resource management goals such as restoration, wildlife habitat enhancement, and invasive species abatement. GPS and GIS tools are invaluable for large scale project design, data collection, and data analysis, with geostatistical methods serving as an alternative or a supplement to the conventional inference statistics in evaluating the project outcome. PMID- 19549298 TI - Primary hepatic embryonal sarcoma masquerading as metastatic ovarian cancer. AB - BACKGROUND: Hepatic embryonal sarcoma (HES) is a rare but aggressive primary tumor of the liver occurring most frequently in childhood. CASE PRESENTATION: We report a case of a 52 year old woman having previously undergone treatment for ovarian serous papillary carcinoma who subsequently presented with a large solitary mass in the liver. Initially this was presumed to be metastasis from the ovarian primary however, on further examination it was shown to be a primary hepatic embryonal sarcoma. CONCLUSION: Primary liver tumors should be considered in differential diagnoses in patients with ovarian cancer who subsequently present with liver tumors. This is particularly important when there is no direct evidence of recurrence of ovarian cancer. PMID- 19549299 TI - Red fluorescent Xenopus laevis: a new tool for grafting analysis. AB - BACKGROUND: Fluorescent proteins such as the green fluorescent protein (GFP) have widely been used in transgenic animals as reporter genes. Their use in transgenic Xenopus tadpoles is especially of interest, because large numbers of living animals can easily be screened. To track more than one event in the same animal, fluorescent markers that clearly differ in their emission spectrum are needed. RESULTS: We established the transgenic Xenopus laevis strain tom3 that expresses ubiquitously red fluorescence from the tdTomato gene through all larval stages and in the adult animal. This new tool was applied to track transplanted blastemas obtained after tail amputation. The blastema can regenerate ectopic tails marked by red fluorescence in the host animal. Surprisingly, we also found contribution of the host animal to form the regenerate. CONCLUSION: We have established a useful new tool to label grafts in Xenopus transplantation experiments. PMID- 19549301 TI - Determinants of weaning success in patients with prolonged mechanical ventilation. AB - INTRODUCTION: Physiological determinants of weaning success and failure are usually studied in ventilator-supported patients, comparing those who failed a trial of spontaneous breathing with those who tolerated such a trial and were successfully extubated. A major limitation of these studies was that the two groups may be not comparable concerning the severity of the underlying disease and the presence of comorbidities. In this physiological study, we assessed the determinants of weaning success in patients acting as their own control, once they are eventually liberated from the ventilator. METHODS: In 30 stable tracheotomised ventilator-dependent patients admitted to a weaning center inside a respiratory intensive care unit, we recorded the breathing pattern, respiratory mechanics, inspiratory muscle function, and tension-time index of diaphragm (TTdi = Pdisw/Pdimax [that is, tidal transdiaphragmatic pressure over maximum transdiaphragmatic pressure] x Ti/Ttot [that is, the inspiratory time over the total breath duration]) at the time of weaning failure (T0). The measurements were repeated in all the patients (T1) either during a successful weaning trial (successful weaning [SW] group, n = 16) or 5 weeks later, in the case of repeated weaning failure (failed weaning [FW] group, n = 14). RESULTS: Compared to T0, in the FW group at T1, significant differences were observed only for a reduction in spontaneous breathing frequency and in TTdi (0.21 +/- 0.122 versus 0.14 +/- 0.054, P = 0.008). SW patients showed a significant increase in Pdimax (34.9 +/- 18.9 cm H2O versus 43.0 +/- 20.0, P = 0.02) and decrease in Pdisw/Pdimax (36.0% +/- 15.8% versus 23.1% +/- 7.9%, P = 0.004). CONCLUSIONS: The recovery of an inadequate inspiratory muscle force could be the major determinant of 'late' weaning success, since this allows the patients to breathe far below the diaphragm fatigue threshold. PMID- 19549302 TI - Pandemic influenza: implications for occupational medicine. AB - This article reviews the biological and occupational medicine literature related to H5N1 pandemic influenza and its impact on infection control, cost and business continuity in settings outside the health care community. The literature on H5N1 biology is reviewed including the treatment and infection control mechanisms as they pertain to occupational medicine. Planning activity for the potential arrival of pandemic avian influenza is growing rapidly. Much has been published on the molecular biology of H5N1 but there remains a paucity of literature on the occupational medicine impacts to organizations. This review summarizes some of the basic science surrounding H5N1 influenza and raises some key concerns in pandemic planning for the occupational medicine professional. Workplaces other than health care settings will be impacted greatly by an H5N1 pandemic and the occupational physician will play an essential role in corporate preparation, response, and business continuity strategies. PMID- 19549300 TI - Results from the national sepsis practice survey: predictions about mortality and morbidity and recommendations for limitation of care orders. AB - INTRODUCTION: Critically ill patients and families rely upon physicians to provide estimates of prognosis and recommendations for care. Little is known about patient and clinician factors which influence these predictions. The association between these predictions and recommendations for continued aggressive care is also understudied. METHODS: We administered a mail-based survey with simulated clinical vignettes to a random sample of the Critical Care Assembly of the American Thoracic Society. Vignettes represented a patient with septic shock with multi-organ failure with identical APACHE II scores and sepsis associated organ failures. Vignettes varied by age (50 or 70 years old), body mass index (BMI) (normal or obese) and co-morbidities (none or recently diagnosed stage IIA lung cancer). All subjects received the vignettes with the highest and lowest mortality predictions from pilot testing and two additional, randomly selected vignettes. Respondents estimated outcomes and selected care for each hypothetical patient. RESULTS: Despite identical severity of illness, the range of estimates for hospital mortality (5th to 95th percentile range, 17% to 78%) and for problems with self-care (5th to 95th percentile range, 2% to 74%) was wide. Similar variation was observed when clinical factors (age, BMI, and co morbidities) were identical. Estimates of hospital mortality and problems with self-care among survivors were significantly higher in vignettes with obese BMIs (4.3% and 5.3% higher, respectively), older age (8.2% and 11.6% higher, respectively), and cancer diagnosis (5.9% and 6.9% higher, respectively). Higher estimates of mortality (adjusted odds ratio 1.29 per 10% increase in predicted mortality), perceived problems with self-care (adjusted odds ratio 1.26 per 10% increase in predicted problems with self-care), and early-stage lung cancer (adjusted odds ratio 5.82) were independently associated with recommendations to limit care. CONCLUSIONS: The studied clinical factors were consistently associated with poorer outcome predictions but did not explain the variation in prognoses offered by experienced physicians. These observations raise concern that provided information and the resulting decisions about continued aggressive care may be influenced by individual physician perception. To provide more reliable and accurate estimates of outcomes, tools are needed which incorporate patient characteristics and preferences with physician predictions and practices. PMID- 19549304 TI - CellMiner: a relational database and query tool for the NCI-60 cancer cell lines. AB - BACKGROUND: Advances in the high-throughput omic technologies have made it possible to profile cells in a large number of ways at the DNA, RNA, protein, chromosomal, functional, and pharmacological levels. A persistent problem is that some classes of molecular data are labeled with gene identifiers, others with transcript or protein identifiers, and still others with chromosomal locations. What has lagged behind is the ability to integrate the resulting data to uncover complex relationships and patterns. Those issues are reflected in full form by molecular profile data on the panel of 60 diverse human cancer cell lines (the NCI-60) used since 1990 by the U.S. National Cancer Institute to screen compounds for anticancer activity. To our knowledge, CellMiner is the first online database resource for integration of the diverse molecular types of NCI-60 and related meta data. DESCRIPTION: CellMiner enables scientists to perform advanced querying of molecular information on NCI-60 (and additional types) through a single web interface. CellMiner is a freely available tool that organizes and stores raw and normalized data that represent multiple types of molecular characterizations at the DNA, RNA, protein, and pharmacological levels. Annotations for each project, along with associated metadata on the samples and datasets, are stored in a MySQL database and linked to the molecular profile data. Data can be queried and downloaded along with comprehensive information on experimental and analytic methods for each data set. A Data Intersection tool allows selection of a list of genes (proteins) in common between two or more data sets and outputs the data for those genes (proteins) in the respective sets. In addition to its role as an integrative resource for the NCI-60, the CellMiner package also serves as a shell for incorporation of molecular profile data on other cell or tissue sample types. CONCLUSION: CellMiner is a relational database tool for storing, querying, integrating, and downloading molecular profile data on the NCI-60 and other cancer cell types. More broadly, it provides a template to use in providing such functionality for other molecular profile data generated by academic institutions, public projects, or the private sector. CellMiner is available online at (http://discover.nci.nih.gov/cellminer/). PMID- 19549303 TI - P-gp activity is a critical resistance factor against AVE9633 and DM4 cytotoxicity in leukaemia cell lines, but not a major mechanism of chemoresistance in cells from acute myeloid leukaemia patients. AB - BACKGROUND: AVE9633 is a new immunoconjugate comprising a humanized monoclonal antibody, anti-CD33 antigen, linked through a disulfide bond to the maytansine derivative DM4, a cytotoxic agent and potent tubulin inhibitor. It is undergoing a phase I clinical trial. Chemoresistance to anti-mitotic agents has been shown to be related, in part, to overexpression of ABC proteins. The aim of the present study was to investigate the potential roles of P-gp, MRP1 and BCRP in cytotoxicity in AVE9633-induced acute myeloid leukaemia (AML). METHODS: This study used AML cell lines expressing different levels of P-gp, MRP1 or BCRP proteins and twenty-five samples from AML patients. Expression and functionality of the transporter protein were analyzed by flow cytometry. The cytotoxicity of the drug was evaluated by MTT and apoptosis assays. RESULTS: P-gp activity, but not MRP1 and BCRP, attenuated AVE9633 and DM4 cytotoxicity in myeloid cell lines. Zosuquidar, a potent specific P-gp inhibitor, restored the sensitivity of cells expressing P-gp to both AVE9633 and DM4. However, the data from AML patients show that 10/25 samples of AML cells (40%) were resistant to AVE9633 or DM4 (IC(50) > 500 nM), and this was not related to P-gp activity (p-Value: 0.7). Zosuquidar also failed to re-establish drug sensitivity. Furthermore, this resistance was not correlated with CD33 expression (p-Value: 0.6) in those cells. CONCLUSION: P gp activity is not a crucial mechanism of chemoresistance to AVE9633. For patients whose resistance to conventional anthracycline AML regimens is related to ABC protein expression, a combination with AVE9633 could be beneficial. Other mechanisms such as microtubule alteration could play an important role in chemoresistance to AVE9633. PMID- 19549305 TI - Alcohol intoxication and mental health among adolescents--a population review of 8983 young people, 13-19 years in North-Trondelag, Norway: the Young-HUNT Study. AB - BACKGROUND: The aims of this study were to describe alcohol use among Norwegian teenagers and investigate the associations between mental health problems and alcohol intoxications with focus on age and gender. METHODS: Population based, cross-sectional survey addressing all adolescents aged 13-19 years, attending secondary or high school in North - Trondelag County, Norway. 8983 youths (91%) answered the Young-HUNT questionnaire in the 1995-1997 survey. Logistic regression models were used to study associations. RESULTS: 80% of the respondents reported that they had tried drinking alcohol, and 57% had been intoxicated at least once. The proportion of the students, which had tried alcohol, was equal in both genders and increased with age. Attention problems and conduct problems were strongly associated with frequent alcohol intoxications in both genders. Anxiety and depressive symptoms among girls were also related to high numbers of intoxications CONCLUSION: Gender differences in number of alcohol intoxications were small. There was a close association between both conduct and attention problems and high alcohol consumption in both genders. Girls with symptoms of anxiety and depression reported more frequent alcohol intoxications. PMID- 19549306 TI - Are we drawing the right conclusions from randomised placebo-controlled trials? A post-hoc analysis of data from a randomised controlled trial. AB - BACKGROUND: Assumptions underlying placebo controlled trials include that the placebo effect impacts on all study arms equally, and that treatment effects are additional to the placebo effect. However, these assumptions have recently been challenged, and different mechanisms may potentially be operating in the placebo and treatment arms. The objective of the current study was to explore the nature of placebo versus pharmacological effects by comparing predictors of the placebo response with predictors of the treatment response in a randomised, placebo controlled trial of a phytotherapeutic combination for the treatment of menopausal symptoms. A substantial placebo response was observed but no significant difference in efficacy between the two arms. METHODS: A post hoc analysis was conducted on data from 93 participants who completed this previously published study. Variables at baseline were investigated as potential predictors of the response on any of the endpoints of flushing, overall menopausal symptoms and depression. Focused tests were conducted using hierarchical linear regression analyses. Based on these findings, analyses were conducted for both groups separately. These findings are discussed in relation to existing literature on placebo effects. RESULTS: Distinct differences in predictors were observed between the placebo and active groups. A significant difference was found for study entry anxiety, and Greene Climacteric Scale (GCS) scores, on all three endpoints. Attitude to menopause was found to differ significantly between the two groups for GCS scores. Examination of the individual arms found anxiety at study entry to predict placebo response on all three outcome measures individually. In contrast, low anxiety was significantly associated with improvement in the active treatment group. None of the variables found to predict the placebo response was relevant to the treatment arm. CONCLUSION: This study was a post hoc analysis of predictors of the placebo versus treatment response. Whilst this study does not explore neurobiological mechanisms, these observations are consistent with the hypotheses that 'drug' effects and placebo effects are not necessarily additive, and that mutually exclusive mechanisms may be operating in the two arms. The need for more research in the area of mechanisms and mediators of placebo versus active responses is supported. TRIAL REGISTRATION: International Clinical Trials Registry ISRCTN98972974. PMID- 19549308 TI - Is inflammation a consequence of extracellular hyperosmolarity? AB - BACKGROUND: There are several reports suggesting that hyperosmolarity induces inflammation. We recently showed that Dextran Sodium Sulfate causes inflammatory bowel disease due to hyperosmolarity. The aim of this study was to confirm the link between hyperosmolarity and inflammation by assessing osmolarity values in vivo during inflammation, compare the inflammatory potential of different osmotic agents and finally study the long-term consequences of hyperosmolarity on cell fate. METHODS: Osmotic pressures were measured in inflammatory liquids withdrawn from mice subjected to inflammation caused either by subcutaneous injection of Bacille Calmette-Guerin (BCG) or Freund adjuvant. Three epithelial cell lines (HT29, T24 and A549) were exposed up to 48 hours to increasing osmolarities (300, 600, 900 mOsm) of chemically inert molecules such as Mannitol, Propylene Glycol, and Glycerol and inflammatory response was assessed by Enzyme Linked ImmunoSorbent Assay (ELISA) and RNA Protection Assay (RPA). Finally, normal mouse macrophages were exposed to hyperosmotic conditions for long-term culture. RESULTS: The inflammation caused either by BCG or Freund adjuvant is correlated to hyperosmolarity in inflammatory liquids. The exposure of cells to the different compounds, whatever their molecular weight, has no effect on the secretion of cytokines as long as the osmolarity is below a threshold of 300 mOsm. Higher osmolarities result in the secretion of proinflammatory cytokines (Interleukin-8, Interleukin-6, Interleukin-1beta and Tumor Necrosis factor alpha). Long-term hyperosmotic culture extends normal macrophage half-life, from 44 days to 102 days, and alters the expression of p53, Bcl-2 and Bax. CONCLUSION: The present study further suggests inflammation and hyperosmolarity are closely related phenomena if not synonymous. PMID- 19549307 TI - Anti-tumor activity of patient-derived NK cells after cell-based immunotherapy--a case report. AB - BACKGROUND: Membrane-bound heat shock protein 70 (Hsp70) serves as a tumor specific recognition structure for Hsp70-peptide (TKD) plus IL-2 activated NK cells. A phase I clinical trial has shown that repeated re-infusions of ex vivo TKD/IL-2-activated, autologous leukapheresis product is safe. This study investigated the maintenance of the cytolytic activity of NK cells against K562 cells and autologous tumor after 6 plus 3 infusions of TKD/IL-2-activated effector cells. METHODS: A stable tumor cell line was generated from the resected anastomotic relapse of a patient with colon carcinoma (pT3, N2, M0, G2). Two months after surgery, the patient received the first monthly i.v. infusion of his ex vivo TKD/IL-2-activated peripheral blood mononuclear cells (PBMNC). After 6 infusions and a pause of 3 months, the patient received another 3 cell infusions. The phenotypic characteristics and activation status of tumor and effector cells were determined immediately before and at times after each infusion. RESULTS: The NK cell ligands Hsp70, MICA/B, and ULBP-1,2,3 were expressed on the patient's anastomotic relapse. An increased density of activatory NK cell receptors following ex vivo stimulation correlated with an enhanced anti-tumoricidal activity. After 4 re-infusion cycles, the intrinsic cytolytic activity of non stimulated PBMNC was significantly elevated and this heightened responsiveness persisted for up to 3 months after the last infusion. Another 2 re-stimulations with TKD/IL-2 restored the cytolytic activity after the therapeutic pause. CONCLUSION: In a patient with colon carcinoma, repeated infusions of ex vivo TKD/IL-2-activated PBMNC initiate an intrinsic NK cell-mediated cytolytic activity against autologous tumor cells. PMID- 19549309 TI - Comparative genomic analysis of C4 photosynthetic pathway evolution in grasses. AB - BACKGROUND: Sorghum is the first C4 plant and the second grass with a full genome sequence available. This makes it possible to perform a whole-genome-level exploration of C4 pathway evolution by comparing key photosynthetic enzyme genes in sorghum, maize (C4) and rice (C3), and to investigate a long-standing hypothesis that a reservoir of duplicated genes is a prerequisite for the evolution of C4 photosynthesis from a C3 progenitor. RESULTS: We show that both whole-genome and individual gene duplication have contributed to the evolution of C4 photosynthesis. The C4 gene isoforms show differential duplicability, with some C4 genes being recruited from whole genome duplication duplicates by multiple modes of functional innovation. The sorghum and maize carbonic anhydrase genes display a novel mode of new gene formation, with recursive tandem duplication and gene fusion accompanied by adaptive evolution to produce C4 genes with one to three functional units. Other C4 enzymes in sorghum and maize also show evidence of adaptive evolution, though differing in level and mode. Intriguingly, a phosphoenolpyruvate carboxylase gene in the C3 plant rice has also been evolving rapidly and shows evidence of adaptive evolution, although lacking key mutations that are characteristic of C4 metabolism. We also found evidence that both gene redundancy and alternative splicing may have sheltered the evolution of new function. CONCLUSIONS: Gene duplication followed by functional innovation is common to evolution of most but not all C4 genes. The apparently long time-lag between the availability of duplicates for recruitment into C4 and the appearance of C4 grasses, together with the heterogeneity of origins of C4 genes, suggests that there may have been a long transition process before the establishment of C4 photosynthesis. PMID- 19549310 TI - Hepatitis C virus core, NS3, NS4B and NS5A are the major immunogenic proteins in humoral immunity in chronic HCV infection. AB - BACKGROUND: The viral genome of hepatitis C virus constitutes a 9.6-kb single stranded positive-sense RNA which encodes altogether 11 viral proteins. In order to study the humoral immune responses against different HCV proteins in patients suffering from chronic HCV infection, we produced three structural (core, E1 and E2) and six nonstructural proteins (NS2, NS3, NS4A, NS4B, NS5A and NS5B) in Sf9 insect cells by using the baculovirus expression system. RESULTS: The recombinant HCV core, E1, E2, NS2, NS3, NS4A, NS4B, NS5A and NS5B proteins were purified and used in Western blot analysis to determine antibody responses against individual HCV protein in 68 HCV RNA and antibody positive human sera that were obtained from patients suffering from genotype 1, 2, 3 or 4 infection. These sera were also analysed with INNO-LIA Score test for HCV antibodies against core, NS3, NS4AB and NS5A, and the results were similar to the ones obtained by Western blot method. Based on our Western blot analyses we found that the major immunogenic HCV antigens were the core, NS4B, NS3 and NS5A proteins which were recognized in 97%, 86%, 68% and 53% of patient sera, respectively. There were no major genotype specific differences in antibody responses to individual HCV proteins. A common feature within the studied sera was that all except two sera recognized the core protein in high titers, whereas none of the sera recognized NS2 protein and only three sera (from genotype 3) recognised NS5B. CONCLUSION: The data shows significant variation in the specificity in humoral immunity in chronic HCV patients. PMID- 19549312 TI - Friction characteristics of Cd-rich carbonate films on calcite surfaces: implications for compositional differentiation at the nanometer scale. AB - Lateral Force Microscopy (LFM) studies were carried out on cleaved calcite sections in contact with solutions supersaturated with respect to otavite (CdCO3) or calcite-otavite solid solutions (SS) as a means to examine the potential for future application of LFM as a nanometer-scale mineral surface composition mapping technique. Layer-by-layer growth of surface films took place either by step advancement or by a surface nucleation and step advancement mechanisms. Friction vs. applied load data acquired on the films and the calcite substrate were successfully fitted to the Johnson Kendall Roberts (JKR) model for single asperity contacts. Following this model, friction differences between film and substrate at low loads were dictated by differences in adhesion, whereas at higher load they reflect differences in contact shear strength. In most experiments at fixed load, the film showed higher friction than the calcite surface, but the friction-load dependence for the different surfaces revealed that at low loads (0-40 nN), a calcian otavite film has lower friction than calcite; a result that is contrary to earlier LFM reports of the same system. Multilayer films of calcian-otavite displayed increasing friction with film thickness, consistent with the expectation that the film surface composition will become increasingly Cd-rich with increasing thickness. Both load- and thickness dependence trends support the hypothesis that the contact shear strength correlates with the hydration enthalpy of the surface ions, thereby imparting friction sensitivity in the LFM to mineral-water interface composition. PMID- 19549311 TI - MLL rearrangements in pediatric acute lymphoblastic and myeloblastic leukemias: MLL specific and lineage specific signatures. AB - BACKGROUND: The presence of MLL rearrangements in acute leukemia results in a complex number of biological modifications that still remain largely unexplained. Armstrong et al. proposed MLL rearrangement positive ALL as a distinct subgroup, separated from acute lymphoblastic (ALL) and myeloblastic leukemia (AML), with a specific gene expression profile. Here we show that MLL, from both ALL and AML origin, share a signature identified by a small set of genes suggesting a common genetic disregulation that could be at the basis of mixed lineage leukemia in both phenotypes. METHODS: Using Affymetrix(R) HG-U133 Plus 2.0 platform, gene expression data from 140 (training set) + 78 (test set) ALL and AML patients with (24+13) and without (116+65) MLL rearrangements have been investigated performing class comparison (SAM) and class prediction (PAM) analyses. RESULTS: We identified a MLL translocation-specific (379 probes) signature and a phenotype specific (622 probes) signature which have been tested using unsupervised methods. A final subset of 14 genes grants the characterization of acute leukemia patients with and without MLL rearrangements. CONCLUSION: Our study demonstrated that a small subset of genes identifies MLL-specific rearrangements and clearly separates acute leukemia samples according to lineage origin. The subset included well-known genes and newly discovered markers that identified ALL and AML subgroups, with and without MLL rearrangements. PMID- 19549313 TI - Development of a biomechanical energy harvester. AB - BACKGROUND: Biomechanical energy harvesting-generating electricity from people during daily activities-is a promising alternative to batteries for powering increasingly sophisticated portable devices. We recently developed a wearable knee-mounted energy harvesting device that generated electricity during human walking. In this methods-focused paper, we explain the physiological principles that guided our design process and present a detailed description of our device design with an emphasis on new analyses. METHODS: Effectively harvesting energy from walking requires a small lightweight device that efficiently converts intermittent, bi-directional, low speed and high torque mechanical power to electricity, and selectively engages power generation to assist muscles in performing negative mechanical work. To achieve this, our device used a one-way clutch to transmit only knee extension motions, a spur gear transmission to amplify the angular speed, a brushless DC rotary magnetic generator to convert the mechanical power into electrical power, a control system to determine when to open and close the power generation circuit based on measurements of knee angle, and a customized orthopaedic knee brace to distribute the device reaction torque over a large leg surface area. RESULTS: The device selectively engaged power generation towards the end of swing extension, assisting knee flexor muscles by producing substantial flexion torque (6.4 Nm), and efficiently converted the input mechanical power into electricity (54.6%). Consequently, six subjects walking at 1.5 m/s generated 4.8 +/- 0.8 W of electrical power with only a 5.0 +/ 21 W increase in metabolic cost. CONCLUSION: Biomechanical energy harvesting is capable of generating substantial amounts of electrical power from walking with little additional user effort making future versions of this technology particularly promising for charging portable medical devices. PMID- 19549315 TI - Characterization and structural analysis of wild type and a non-abscission mutant at the development funiculus (Def) locus in Pisum sativum L. AB - BACKGROUND: In pea seeds (Pisum sativum L.), the Def locus defines an abscission event where the seed separates from the funicle through the intervening hilum region at maturity. A spontaneous mutation at this locus results in the seed failing to abscise from the funicle as occurs in wild type peas. In this work, structural differences between wild type peas that developed a distinct abscission zone (AZ) between the funicle and the seed coat and non-abscission def mutant were characterized. RESULTS: A clear abscission event was observed in wild type pea seeds that were associated with a distinct double palisade layers at the junction between the seed coat and funicle. Generally, mature seeds fully developed an AZ, which was not present in young wild type seeds. The AZ was formed exactly below the counter palisade layer. In contrast, the palisade layers at the junction of the seed coat and funicle were completely absent in the def mutant pea seeds and the cells in this region were seen to be extensions of surrounding parenchymatous cells. CONCLUSION: The Def wild type developed a distinct AZ associated with palisade layer and counterpalisade layer at the junction of the seed coat and funicle while the def mutant pea seed showed non abscission and an absence of the double palisade layers in the same region. We conclude that the presence of the double palisade layer in the hilum of the wild type pea seeds plays an important structural role in AZ formation by delimiting the specific region between the seed coat and the funicle and may play a structural role in the AZ formation and subsequent detachment of the seed from the funicle. PMID- 19549314 TI - Degradome expression profiling in human articular cartilage. AB - INTRODUCTION: The molecular mechanisms underlying cartilage destruction in osteoarthritis are poorly understood. Proteolysis is a key feature in the turnover and degradation of cartilage extracellular matrix where the focus of research has been on the metzincin family of metalloproteinases. However, there is strong evidence to indicate important roles for other catalytic classes of proteases, with both extracellular and intracellular activities. The aim of this study was to profile the expression of the majority of protease genes in all catalytic classes in normal human cartilage and that from patients with osteoarthritis (OA) using a quantitative method. METHODS: Human cartilage was obtained from femoral heads at joint replacement for either osteoarthritis or following fracture to the neck of femur (NOF). Total RNA was purified, and expression of genes assayed using Taqman low-density array quantitative RT-PCR. RESULTS: A total of 538 protease genes were profiled, of which 431 were expressed in cartilage. A total of 179 genes were differentially expressed in OA versus NOF cartilage: eight aspartic proteases, 44 cysteine proteases, 76 metalloproteases, 46 serine proteases and five threonine proteases. Wilcoxon ranking as well as the LogitBoost-NR machine learning approach were used to assign significance to each gene, with the most highly ranked genes broadly similar using each method. CONCLUSIONS: This study is the most complete quantitative analysis of protease gene expression in cartilage to date. The data help give direction to future research on the specific function(s) of individual proteases or protease families in cartilage and may help to refine anti-proteolytic strategies in OA. PMID- 19549316 TI - Analysis of von Willebrand factor A domain-related protein (WARP) polymorphism in temperate and tropical Plasmodium vivax field isolates. AB - BACKGROUND: The identification of key molecules is crucial for designing transmission-blocking vaccines (TBVs), among those ookinete micronemal proteins are candidate as a general class of malaria transmission-blocking targets. Here, the sequence analysis of an extra-cellular malaria protein expressed in ookinetes, named von Willebrand factor A domain-related protein (WARP), is reported in 91 Plasmodium vivax isolates circulating in different regions of Iran. METHODS: Clinical isolates were collected from north temperate and southern tropical regions in Iran. Primers have been designed based on P. vivax sequence (ctg_6991) which amplified a fragment of about 1044 bp with no size variation. Direct sequencing of PCR products was used to determine polymorphism and further bioinformatics analysis in P. vivax sexual stage antigen, pvwarp. RESULTS: Amplified pvwarp gene showed 886 bp in size, with no intron. BLAST analysis showed a similarity of 98-100% to P. vivax Sal-I strain; however, Iranian isolates had 2 bp mismatches in 247 and 531 positions that were non-synonymous substitution [T (ACT) to A (GCT) and R (AGA) to S (AGT)] in comparison with the Sal-I sequence. CONCLUSION: This study presents the first large-scale survey on pvwarp polymorphism in the world, which provides baseline data for developing WARP-based TBV against both temperate and tropical P. vivax isolates. PMID- 19549317 TI - Physiologic upper limit of pore size in the blood-tumor barrier of malignant solid tumors. AB - BACKGROUND: The existence of large pores in the blood-tumor barrier (BTB) of malignant solid tumor microvasculature makes the blood-tumor barrier more permeable to macromolecules than the endothelial barrier of most normal tissue microvasculature. The BTB of malignant solid tumors growing outside the brain, in peripheral tissues, is more permeable than that of similar tumors growing inside the brain. This has been previously attributed to the larger anatomic sizes of the pores within the BTB of peripheral tumors. Since in the physiological state in vivo a fibrous glycocalyx layer coats the pores of the BTB, it is possible that the effective physiologic pore size in the BTB of brain tumors and peripheral tumors is similar. If this were the case, then the higher permeability of the BTB of peripheral tumor would be attributable to the presence of a greater number of pores in the BTB of peripheral tumors. In this study, we probed in vivo the upper limit of pore size in the BTB of rodent malignant gliomas grown inside the brain, the orthotopic site, as well as outside the brain in temporalis skeletal muscle, the ectopic site. METHODS: Generation 5 (G5) through generation 8 (G8) polyamidoamine dendrimers were labeled with gadolinium (Gd) diethyltriaminepentaacetic acid, an anionic MRI contrast agent. The respective Gd dendrimer generations were visualized in vitro by scanning transmission electron microscopy. Following intravenous infusion of the respective Gd-dendrimer generations (Gd-G5, N = 6; Gd-G6, N = 6; Gd-G7, N = 5; Gd-G8, N = 5) the blood and tumor tissue pharmacokinetics of the Gd-dendrimer generations were visualized in vivo over 600 to 700 minutes by dynamic contrast-enhanced MRI. One additional animal was imaged in each Gd-dendrimer generation group for 175 minutes under continuous anesthesia for the creation of voxel-by-voxel Gd concentration maps. RESULTS: The estimated diameters of Gd-G7 dendrimers were 11 +/- 1 nm and those of Gd-G8 dendrimers were 13 +/- 1 nm. The BTB of ectopic RG-2 gliomas was more permeable than the BTB of orthotopic RG-2 gliomas to all Gd-dendrimer generations except for Gd-G8. The BTB of both ectopic RG-2 gliomas and orthotopic RG-2 gliomas was not permeable to Gd-G8 dendrimers. CONCLUSION: The physiologic upper limit of pore size in the BTB of malignant solid tumor microvasculature is approximately 12 nanometers. In the physiologic state in vivo the luminal fibrous glycocalyx of the BTB of malignant brain tumor and peripheral tumors is the primary impediment to the effective transvascular transport of particles across the BTB of malignant solid tumor microvasculature independent of tumor host site. The higher permeability of malignant peripheral tumor microvasculature to macromolecules smaller than approximately 12 nm in diameter is attributable to the presence of a greater number of pores underlying the glycocalyx of the BTB of malignant peripheral tumor microvasculature. PMID- 19549318 TI - OrthoClusterDB: an online platform for synteny blocks. AB - BACKGROUND: The recent availability of an expanding collection of genome sequences driven by technological advances has facilitated comparative genomics and in particular the identification of synteny among multiple genomes. However, the development of effective and easy-to-use methods for identifying such conserved gene clusters among multiple genomes-synteny blocks-as well as databases, which host synteny blocks from various groups of species (especially eukaryotes) and also allow users to run synteny-identification programs, lags behind. DESCRIPTIONS: OrthoClusterDB is a new online platform for the identification and visualization of synteny blocks. OrthoClusterDB consists of two key web pages: Run OrthoCluster and View Synteny. The Run OrthoCluster page serves as web front-end to OrthoCluster, a recently developed program for synteny block detection. Run OrthoCluster offers full control over the functionalities of OrthoCluster, such as specifying synteny block size, considering order and strandedness of genes within synteny blocks, including or excluding nested synteny blocks, handling one-to-many orthologous relationships, and comparing multiple genomes. In contrast, the View Synteny page gives access to perfect and imperfect synteny blocks precomputed for a large number of genomes, without the need for users to retrieve and format input data. Additionally, genes are cross linked with public databases for effective browsing. For both Run OrthoCluster and View Synteny, identified synteny blocks can be browsed at the whole genome, chromosome, and individual gene level. OrthoClusterDB is freely accessible. CONCLUSION: We have developed an online system for the identification and visualization of synteny blocks among multiple genomes. The system is freely available at (http://genome.sfu.ca/orthoclusterdb/). PMID- 19549319 TI - Conflicting priorities: evaluation of an intervention to improve nurse-parent relationships on a Tanzanian paediatric ward. AB - BACKGROUND: Patient, or parent/guardian, satisfaction with health care provision is important to health outcomes. Poor relationships with health workers, particularly with nursing staff, have been reported to reduce satisfaction with care in Africa. Participatory research approaches such as the Health Workers for Change initiative have been successful in improving provider-client relationships in various developing country settings, but have not yet been reported in the complex environment of hospital wards. We evaluated the HWC approach for improving the relationship between nurses and parents on a paediatric ward in a busy regional hospital in Tanzania. METHODS: The intervention consisted of six workshops, attended by 29 of 31 trained nurses and nurse attendants working on the paediatric ward. Parental satisfaction with nursing care was measured with 288 parents before and six weeks after the workshops, by means of an adapted Picker questionnaire. Two focus-group discussions were held with the workshop participants six months after the intervention. RESULTS: During the workshops, nurses demonstrated awareness of poor relationships between themselves and mothers. To tackle this, they proposed measures including weekly meetings to solve problems, maintain respect and increase cooperation, and representation to administrative forces to request better working conditions such as equipment, salaries and staff numbers. The results of the parent satisfaction questionnaire showed some improvement in responsiveness of nurses to client needs, but overall the mean percentage of parents reporting each of 20 problems was not statistically significantly different after the intervention, compared to before it (38.9% versus 41.2%). Post-workshop focus-group discussions with nursing staff suggested that nurses felt more empathic towards mothers and perceived an improvement in the relationship, but that this was hindered by persisting problems in their working environment, including poor relationships with other staff and a lack of response from hospital administration to their needs. CONCLUSION: The intended outcome of the intervention was not met. The priorities of the intervention--to improve nurse-parent relationships--did not match the priorities of the nursing staff. Development of awareness and empathy was not enough to provide care that was satisfactory to clients in the context of working conditions that were unsatisfactory to nurses. PMID- 19549320 TI - Carbon and arsenic metabolism in Thiomonas strains: differences revealed diverse adaptation processes. AB - BACKGROUND: Thiomonas strains are ubiquitous in arsenic-contaminated environments. Differences between Thiomonas strains in the way they have adapted and respond to arsenic have never been studied in detail. For this purpose, five Thiomonas strains, that are interesting in terms of arsenic metabolism were selected: T. arsenivorans, Thiomonas spp. WJ68 and 3As are able to oxidise As(III), while Thiomonas sp. Ynys1 and T. perometabolis are not. Moreover, T. arsenivorans and 3As present interesting physiological traits, in particular that these strains are able to use As(III) as an electron donor. RESULTS: The metabolism of carbon and arsenic was compared in the five Thiomonas strains belonging to two distinct phylogenetic groups. Greater physiological differences were found between these strains than might have been suggested by 16S rRNA/rpoA gene phylogeny, especially regarding arsenic metabolism. Physiologically, T. perometabolis and Ynys1 were unable to oxidise As(III) and were less arsenic resistant than the other strains. Genetically, they appeared to lack the aox arsenic-oxidising genes and carried only a single ars arsenic resistance operon. Thiomonas arsenivorans belonged to a distinct phylogenetic group and increased its autotrophic metabolism when arsenic concentration increased. Differential proteomic analysis revealed that in T. arsenivorans, the rbc/cbb genes involved in the assimilation of inorganic carbon were induced in the presence of arsenic, whereas these genes were repressed in Thiomonas sp. 3As. CONCLUSION: Taken together, these results show that these closely related bacteria differ substantially in their response to arsenic, amongst other factors, and suggest different relationships between carbon assimilation and arsenic metabolism. PMID- 19549321 TI - Full-length cytokeratin-19 is released by human tumor cells: a potential role in metastatic progression of breast cancer. AB - INTRODUCTION: We evaluated whether CK19, one of the main cytoskeleton proteins of epithelial cells, is released as full-length protein from viable tumor cells and whether this property is relevant for metastatic progression in breast cancer patients. METHODS: EPISPOT (EPithelial ImmunoSPOT) assays were performed to analyze the release of full-length CK19 by carcinoma cells of various origins, and the sequence of CK19 was analyzed with mass spectrometry. Additional functional experiments with cycloheximide, Brefeldin A, or vincristine were done to analyze the biology of the CK19-release. CK19-EPISPOT was used to detect disseminated tumor cells in bone marrow (BM) of 45 breast cancer patients who were then followed up over a median of 6 years. RESULTS: CK19 was expressed and released by colorectal (HT-29, HCT116, Caco-2) and breast (MCF-7, SKBR3, and MDA MB-231) cancer cell lines. The CK19-EPISPOT was more sensitive than the CK19 ELISA. Dual fluorescent EPISPOT with antibodies against different CK19 epitopes showed the release of the full-length CK19, which was confirmed by mass spectrometry. Functional experiments indicated that CK19 release was an active process and not simply the consequence of cell death. CK19-releasing cells (RCs) were detectable in BM of 44% to 70% of breast cancer patients. This incidence and the number of CK19-RCs were correlated to the presence of overt metastases, and patients with CK19-RCs had a reduced survival as compared with patients without these cells (P = 0.025, log-rank test; P = 0.0019, hazard ratio, 4.7; multivariate analysis). CONCLUSIONS: Full-length CK19 is released by viable epithelial tumor cells, and CK19-RCs might constitute a biologically active subset of breast cancer cells with high metastatic properties. PMID- 19549322 TI - Determining the date of diagnosis--is it a simple matter? The impact of different approaches to dating diagnosis on estimates of delayed care for ovarian cancer in UK primary care. AB - BACKGROUND: Studies of cancer incidence and early management will increasingly draw on routine electronic patient records. However, data may be incomplete or inaccurate. We developed a generalizable strategy for investigating presenting symptoms and delays in diagnosis using ovarian cancer as an example. METHODS: The General Practice Research Database was used to investigate the time between first report of symptom and diagnosis of 344 women diagnosed with ovarian cancer between 01/06/2002 and 31/05/2008. Effects of possible inaccuracies in dating of diagnosis on the frequencies and timing of the most commonly reported symptoms were investigated using four increasingly inclusive definitions of first diagnosis/suspicion: 1. "Definite diagnosis" 2. "Ambiguous diagnosis" 3. "First treatment or complication suggesting pre-existing diagnosis", 4 "First relevant test or referral". RESULTS: The most commonly coded symptoms before a definite diagnosis of ovarian cancer, were abdominal pain (41%), urogenital problems(25%), abdominal distension (24%), constipation/change in bowel habits (23%) with 70% of cases reporting at least one of these. The median time between first reporting each of these symptoms and diagnosis was 13, 21, 9.5 and 8.5 weeks respectively. 19% had a code for definitions 2 or 3 prior to definite diagnosis and 73% a code for 4. However, the proportion with symptoms and the delays were similar for all four definitions except 4, where the median delay was 8, 8, 3, 10 and 0 weeks respectively. CONCLUSION: Symptoms recorded in the General Practice Research Database are similar to those reported in the literature, although their frequency is lower than in studies based on self-report. Generalizable strategies for exploring the impact of recording practice on date of diagnosis in electronic patient records are recommended, and studies which date diagnoses in GP records need to present sensitivity analyses based on investigation, referral and diagnosis data. Free text information may be essential in obtaining accurate estimates of incidence, and for accurate dating of diagnoses. PMID- 19549323 TI - Cross-fostering does not alter the neurochemistry or behavior of spontaneously hypertensive rats. AB - BACKGROUND: Attention-deficit/hyperactivity disorder (ADHD) is a highly heritable developmental disorder resulting from complex gene-gene and gene-environment interactions. The most widely used animal model, the spontaneously hypertensive rat (SHR), displays the major symptoms of ADHD (deficits in attention, impulsivity and hyperactivity) and has a disturbance in the noradrenergic system when compared to control Wistar-Kyoto rats (WKY). The aim of the present study was to determine whether the ADHD-like characteristics of SHR were purely genetically determined or dependent on the gene-environment interaction provided by the SHR dam. METHODS: SHR/NCrl (Charles River, USA), WKY/NCrl (Charles River, USA) and Sprague Dawley rats (SD/Hsd, Harlan, UK) were bred at the University of Cape Town. Rat pups were cross-fostered on postnatal day 2 (PND 2). Control rats remained with their birth mothers to serve as a reference for their particular strain phenotype. Behavior in the open-field and the elevated-plus maze was assessed between PND 29 and 33. Two days later, rats were decapitated and glutamate-stimulated release of [3H]norepinephrine was determined in prefrontal cortex and hippocampal slices. RESULTS: There was no significant effect of "strain of dam" but there was a significant effect of "pup strain" on all parameters investigated. SHR pups travelled a greater distance in the open field, spent a longer period of time in the inner zone and entered the inner zone of the open-field more frequently than SD or WKY. SD were more active than WKY in the open-field. WKY took longer to enter the inner zone than SHR or SD. In the elevated-plus maze, SHR spent less time in the closed arms, more time in the open arms and entered the open arms more frequently than SD or WKY. There was no difference between WKY and SD behavior in the elevated-plus maze. SHR released significantly more [3H]norepinephrine in response to glutamate than SD or WKY in both hippocampus and prefrontal cortex while SD prefrontal cortex released more [3H]norepinephrine than WKY. SHR were resilient, cross-fostering did not reduce their ADHD-like behavior or change their neurochemistry. Cross-fostering of SD pups onto SHR or WKY dams increased their exploratory behavior without altering their anxiety-like behavior. CONCLUSION: The ADHD-like behavior of SHR and their neurochemistry is genetically determined and not dependent on nurturing by SHR dams. The similarity between WKY and SD supports the continued use of WKY as a control for SHR and suggests that SD may be a useful additional reference strain for SHR. The fact that SD behaved similarly to WKY in the elevated-plus maze argues against the use of WKY as a model for anxiety-like disorders. PMID- 19549324 TI - Investigations of fine-scale phylogeography in Tigriopus californicus reveal historical patterns of population divergence. AB - BACKGROUND: The intertidal copepod Tigriopus californicus is a model for studying the process of genetic divergence in allopatry and for probing the nature of genetic changes that lead to reproductive isolation. Although previous studies have revealed a pattern of remarkably high levels of genetic divergence between the populations of this species at several spatial scales, it is not clear what types of historical processes are responsible. Particularly lacking are data that can yield insights into population history from the finest scales of geographic resolution. RESULTS: Sequence variation in both cytochrome b (CYTB, mtDNA) and the rieske iron-sulfur protein (RISP, nuclear) are examined at a fine scale within four different regions for populations of T. californicus. High levels of genetic divergence are seen for both genes at the broader scale, and genetic subdivision is apparent at nearly all scales in these populations for these two genes. Patterns of polymorphism and divergence in both CYTB and RISP suggest that selection may be leading to non-neutral evolution of these genes in several cases but a pervasive pattern of neither selection nor coadaptation is seen for these markers. CONCLUSION: The use of sequence data at a fine-scale of resolution in this species has provided novel insights into the processes that have resulted in the accumulation of genetic divergence among populations. This divergence is likely to result from an interplay between a limited dispersal ability for this copepod and the temporal instability of copepod habitat. Both shorter-term processes such as the extinction/recolonization dynamics of copepod pools and longer-term processes such as geological uplift of coastline and sea level changes appear to have impacted the patterns of differentiation. Some patterns of sequence variation are consistent with selection acting upon the loci used in this study; however, it appears that most phylogeographic patterns are the result of history and not selection on these genes in this species. PMID- 19549325 TI - Out-of-hours care in western countries: assessment of different organizational models. AB - BACKGROUND: Internationally, different organizational models are used for providing out-of-hours care. The aim of this study was to assess prevailing models in order to identify their potential strengths and weaknesses. METHODS: An international web-based survey was done in 2007 in a sample of purposefully selected key informants from 25 western countries. The questions concerned prevailing organizational models for out-of-hours care, the most dominant model in each country, perceived weaknesses, and national plans for changes in out-of hours care. RESULTS: A total of 71 key informants from 25 countries provided answers. In most countries several different models existed alongside each other. The Accident and Emergency department was the organizational model most frequently used. Perceived weaknesses of this model concerned the coordination and continuity of care, its efficiency and accessibility. In about a third of the countries, the rota group was the most dominant organizational model for out-of hours care. A perceived weakness of this model was lowered job satisfaction of physicians. The GP cooperative existed in a majority of the participating countries; no weaknesses were mentioned with respect to this model. Most of the countries had plans to change the out-of-hours care, mainly toward large scale organizations. CONCLUSION: GP cooperatives combine size of scale advantages with organizational features of strong primary care, such as high accessibility, continuity and coordination of care. While specific patients require other organizational models, the co-existence of different organizational models for out-of-hours care in a country may be less efficient for health systems. PMID- 19549327 TI - The effects of spatial and temporal heterogeneity on the population dynamics of four animal species in a Danish landscape. AB - BACKGROUND: Variation in carrying capacity and population return rates is generally ignored in traditional studies of population dynamics. Variation is hard to study in the field because of difficulties controlling the environment in order to obtain statistical replicates, and because of the scale and expense of experimenting on populations. There may also be ethical issues. To circumvent these problems we used detailed simulations of the simultaneous behaviours of interacting animals in an accurate facsimile of a real Danish landscape. The models incorporate as much as possible of the behaviour and ecology of skylarks Alauda arvensis, voles Microtus agrestis, a ground beetle Bembidion lampros and a linyphiid spider Erigone atra. This allows us to quantify and evaluate the importance of spatial and temporal heterogeneity on the population dynamics of the four species. RESULTS: Both spatial and temporal heterogeneity affected the relationship between population growth rate and population density in all four species. Spatial heterogeneity accounted for 23-30% of the variance in population growth rate after accounting for the effects of density, reflecting big differences in local carrying capacity associated with the landscape features important to individual species. Temporal heterogeneity accounted for 3-13% of the variance in vole, skylark and spider, but 43% in beetles. The associated temporal variation in carrying capacity would be problematic in traditional analyses of density dependence. Return rates were less than one in all species and essentially invariant in skylarks, spiders and beetles. Return rates varied over the landscape in voles, being slower where there were larger fluctuations in local population sizes. CONCLUSION: Our analyses estimated the traditional parameters of carrying capacities and return rates, but these are now seen as varying continuously over the landscape depending on habitat quality and the mechanisms of density dependence. The importance of our results lies in our demonstration that the effects of spatial and temporal heterogeneity must be accounted for if we are to have accurate predictive models for use in management and conservation. This is an area which until now has lacked an adequate theoretical framework and methodology. PMID- 19549326 TI - Neuroendocrine transcriptional programs adapt dynamically to the supply and demand for neuropeptides as revealed in NSF mutant zebrafish. AB - BACKGROUND: Regulated secretion of specialized neuropeptides in the vertebrate neuroendocrine system is critical for ensuring physiological homeostasis. Expression of these cell-specific peptide markers in the differentiating hypothalamus commences prior to birth, often predating the physiological demand for secreted neuropeptides. The conserved function and spatial expression of hypothalamic peptides in vertebrates prompted us to search for critical neuroendocrine genes in newly hatched zebrafish larvae. RESULTS: We screened mutant 5 days post-fertilization zebrafish larvae that fail to undergo visually mediated background adaptation for disruption in hypothalamic pomc expression. To our surprise, the ATPase N-ethylmaleimide sensitive factor (nsf) was identified as an essential gene for maintenance of neuroendocrine transcriptional programs during the embryo-to-larva transition. Despite normal hypothalamic development in nsf(st53) mutants, neuropeptidergic cells exhibited a dramatic loss of cell specific markers by 5 days post-fertilization that is accompanied by elevated intracellular neuropeptide protein. Consistent with the role of NSF in vesicle membrane fusion events and intracellular trafficking, cytoplasmic endoplasmic reticulum-like membranes accumulate in nsf(-/-) hypothalamic neurons similar to that observed for SEC18 (nsf ortholog) yeast mutants. Our data support a model in which unspent neuropeptide cargo feedbacks to extinguish transcription in neuropeptidergic cells just as they become functionally required. In support of this model we found that gnrh3 transcripts remained unchanged in pre-migratory, non-functional gonadotropin-releasing hormone (GnRH) neurons in nsf(-/-) zebrafish. Furthermore, oxytocin-like (oxtl, intp) transcripts, which are found in osmoreceptive neurons and persist in mutant zebrafish, drop precipitously after mutant zebrafish are acutely challenged with high salt. CONCLUSION: Our analyses of nsf mutant zebrafish reveal an unexpected role for NSF in hypothalamic development, with mutant 5 days post-fertilization larvae exhibiting a stage-dependent loss of neuroendocrine transcripts and a corresponding accumulation of neuropeptides in the soma. Based on our collective findings, we speculate that neuroendocrine transcriptional programs adapt dynamically to both the supply and demand for neuropeptides to ensure adequate homeostatic responses. PMID- 19549328 TI - Expression of FOXA1 and GATA-3 in breast cancer: the prognostic significance in hormone receptor-negative tumours. AB - INTRODUCTION: The expression of additional genes, other than oestrogen receptor (ER), may be important to the hormone-responsive phenotype of breast cancer. Microarray analyses have revealed that forkhead box A1 (FOXA1) and GATA binding protein 3 (GATA-3) are expressed in close association with ERalpha, both encoding for transcription factors with a potential involvement in the ERalpha-mediated action in breast cancer. The purpose of this study was to explore if the expression of FOXA1 and GATA-3 may provide an opportunity to stratify subsets of patients that could have better outcome, among the ERalpha-negative/poor prognosis breast cancer group. METHODS: We evaluate FOXA1 and GATA-3 expression in 249 breast carcinomas by immunohistochemistry, associating it with breast cancer molecular markers, clinicopathological features and patient's survival. The clinicopathological features and immunohistochemical markers of the tumours were compared using the chi-square test and ANOVA. Disease-free survival was analysed through Kaplan-Meier survival curves and Cox regression. RESULTS: FOXA1 expression was demonstrated in 42% of invasive carcinomas, while GATA-3 was detected in 48% of the cases. FOXA1 expression was inversely associated with tumour size, Nottingham Prognostic Index, histological grade, lymph vascular invasion, lymph node stage and human epidermal growth factor receptor-2 (HER-2) overexpression, while GATA-3 expression showed inverse association with histological grade and HER-2. Both FOXA1 and GATA-3 were directly associated with ERalpha and progesterone receptor. Among FOXA1-positive tumours, 83.1% are comprised in the luminal A subtype, similar to GATA-3 where 87.7% of positive tumours were classified within this molecular subtype. In the subset of ERalpha negative patients, those who were FOXA1-negative had a 3.61-fold increased risk of breast cancer recurrence when compared with the FOXA1-positive. CONCLUSIONS: FOXA1 was a significant predictor of good outcome in breast cancer, whereas GATA 3 was an important luminal marker. The expression of FOXA1 may be used for risk stratification among ERalpha-negative patients. PMID- 19549329 TI - Timing of surgical antibiotic prophylaxis administration: complexities of analysis. AB - BACKGROUND: The timing of prophylactic antibiotic administration is a patient safety outcome that is recurrently tracked and reported. The interpretation of these data has important implications for patient safety practices. However, diverse data collection methods and approaches to analysis impede knowledge building in this field. This paper makes explicit several challenges to quantifying the timing of prophylactic antibiotics that we encountered during a recent study and offers a suggested protocol for resolving these challenges. CHALLENGES: Two clear challenges manifested during the data extraction process: the actual classification of antibiotic timing, and the additional complication of multiple antibiotic regimens with different timing classifications in a single case. A formalized protocol was developed for dealing with incomplete, ambiguous and unclear documentation. A hierarchical coding system was implemented for managing cases with multiple antibiotic regimens. INTERPRETATION: Researchers who are tracking prophylactic antibiotic timing as an outcome measure should be aware that documentation of antibiotic timing in the patient chart is frequently incomplete and unclear, and these inconsistencies should be accounted for in analyses. We have developed a systematic method for dealing with specific problematic patterns encountered in the data. We propose that the general adoption of a systematic approach to analysis of this type of data will allow for cross-study comparisons and ensure that interpretation of results is on the basis of timing practices rather than documentation practices. PMID- 19549330 TI - Cytometric detection of antigen-specific IFN-gamma/IL-2 secreting cells in the diagnosis of tuberculosis. AB - BACKGROUND: The purpose of this study was to further characterize the immune response to Mycobacterium tuberculosis (Mtb) antigens, in order to provide new insight into host-pathogen interactions in tuberculosis (TB), and to offer tools for a more accurate diagnosis of the different stages of TB. METHODS: T-cell responses to Bacillus Calmette-Guerin (BCG), purified protein derivative (PPD), early secretory antigenic target-6 (ESAT-6) protein and culture filtrate protein 10 kDa (CFP-10) were measured in terms of interferon (IFN)-gamma and interleukin (IL)-2 release, using a novel flow cytometric cell-secreting cytokine detection technique. The study was conducted on peripheral blood mononuclear cells (PBMC) obtained from active TB patients, latently TB infected individuals, and healthy donors. IL-10 and IL-17 were also measured to test their possible role as indicators of disease activity. RESULTS: We confirmed that the enumeration of IFN gamma releasing cells upon Mtb-specific stimulation is sufficient to identify TB patients and that CD8+ T cells concur to IFN-gamma secretion. IL-2 secreting cells were more frequently observed in latent TB infected individuals compared to active TB patients, suggesting that measurement of cells secreting this cytokine could be a marker of disease stage. No discriminating role was associated to IL 10 and IL-17 release in TB patients. CONCLUSION: Our data indicate that the flow cytometric cytokine-secreting cell detection technique may be envisaged as an additional tool for TB diagnosis allowing the analysis of the immune response to M. tuberculosis-related antigens in the different stages of TB. PMID- 19549331 TI - Interim data monitoring to enroll higher-risk participants in HIV prevention trials. AB - BACKGROUND: Lower-than-expected incidence of HIV undermines sample size calculations and compromises the power of a HIV prevention trial. We evaluated the effectiveness of interim monitoring of HIV infection rates and on-going modification of recruitment strategies to enroll women at higher risk of HIV in the Cellulose Sulfate Phase III study in Nigeria. METHODS: We analyzed prevalence and incidence of HIV and other sexually transmitted infections, demographic and sexual behavior characteristics aggregated over the treatment groups on a quarterly basis. The site investigators were advised on their recruitment strategies based on the findings of the interim analyses. RESULTS: A total of 3619 women were screened and 1644 enrolled at the Ikeja and Apapa clinics in Lagos, and at the Central and Peripheral clinics in Port Harcourt. Twelve months after study initiation, the overall incidence of HIV was less than one-third of the pre-study assumption, with rates of HIV that varied substantially between clinics. Due to the low prevalence and incidence rates of HIV, it was decided to close the Ikeja clinic in Lagos and to find new catchment areas in Port Harcourt. This strategy was associated with an almost two-fold increase in observed HIV incidence during the second year of the study. CONCLUSION: Given the difficulties in estimating HIV incidence, a close monitoring of HIV prevalence and incidence rates during a trial is warranted. The on-going modification of recruitment strategies based on the regular analysis of HIV rates appeared to be an efficient method for targeting populations at greatest risk of HIV infection and increasing study power in the Nigeria trial. TRIAL REGISTRATION: The trial was registered with the ClinicalTrials.gov registry under #NCT00120770 http://clinicaltrials.gov/ct2/show/NCT00120770. PMID- 19549332 TI - Diversity and evolution of the small multidrug resistance protein family. AB - BACKGROUND: Members of the small multidrug resistance (SMR) protein family are integral membrane proteins characterized by four alpha-helical transmembrane strands that confer resistance to a broad range of antiseptics and lipophilic quaternary ammonium compounds (QAC) in bacteria. Due to their short length and broad substrate profile, SMR proteins are suggested to be the progenitors for larger alpha-helical transporters such as the major facilitator superfamily (MFS) and drug/metabolite transporter (DMT) superfamily. To explore their evolutionary association with larger multidrug transporters, an extensive bioinformatics analysis of SMR sequences (> 300 Bacteria taxa) was performed to expand upon previous evolutionary studies of the SMR protein family and its origins. RESULTS: A thorough annotation of unidentified/putative SMR sequences was performed placing sequences into each of the three SMR protein subclass designations, namely small multidrug proteins (SMP), suppressor of groEL mutations (SUG), and paired small multidrug resistance (PSMR) using protein alignments and phylogenetic analysis. Examination of SMR subclass distribution within Bacteria and Archaea taxa identified specific Bacterial classes that uniquely encode for particular SMR subclass members. The extent of selective pressure acting upon each SMR subclass was determined by calculating the rate of synonymous to non synonymous nucleotide substitutions using Syn-SCAN analysis. SUG and SMP subclasses are maintained under moderate selection pressure in comparison to integron and plasmid encoded SMR homologues. Conversely, PSMR sequences are maintained under lower levels of selection pressure, where one of the two PSMR pairs diverges in sequence more rapidly than the other. SMR genomic loci surveys identified potential SMR efflux substrates based on its gene association to putative operons that encode for genes regulating amino acid biogenesis and QAC like metabolites. SMR subclass protein transmembrane domain alignments to Bacterial/Archaeal transporters (BAT), DMT, and MFS sequences supports SMR participation in multidrug transport evolution by identifying common TM domains. CONCLUSION: Based on this study, PSMR sequences originated recently within both SUG and SMP clades through gene duplication events and it appears that SMR members may be evolving towards specific metabolite transport. PMID- 19549333 TI - Low-dose vasopressin infusion results in increased mortality and cardiac dysfunction following ischemia-reperfusion injury in mice. AB - INTRODUCTION: Arginine vasopressin is a vasoactive drug commonly used in distributive shock states including mixed shock with a cardiac component. However, the direct effect of arginine vasopressin on the function of the ischemia/reperfusion injured heart has not been clearly elucidated. METHODS: We measured left ventricular ejection fraction using trans-thoracic echocardiography in C57B6 mice, both in normal controls and following ischemia/reperfusion injury induced by a one hour ligation of the left anterior descending coronary artery. Mice were treated with one of normal saline, dobutamine (8.33 microg/kg/min), or arginine vasopressin (0.00057 Units/kg/min, equivalent to 0.04 Units/min in a 70 kg human) delivered by an intraperitoneal micro-osmotic pump. Arterial blood pressure was measured using a micromanometer catheter. In addition, mortality was recorded and cardiac tissues processed for RNA and protein. RESULTS: Baseline left ventricular ejection fraction was 65.6% (60 to 72). In normal control mice, there was no difference in left ventricular ejection fraction according to infusion group. Following ischemia/reperfusion injury, AVP treatment significantly reduced day 1 left ventricular ejection fraction 46.2% (34.4 to 52.0), both in comparison with baseline and day 1 saline treated controls 56.9% (42.4 to 60.2). There were no significant differences in preload (left ventricular end diastolic volume), afterload (blood pressure) or heart rate to account for the effect of AVP on left ventricular ejection fraction. The seven day mortality rate was highest in the arginine vasopressin group. Following ischemia/reperfusion injury, we found no change in cardiac V1 Receptor expression but a 40% decrease in Oxytocin Receptor expression. CONCLUSIONS: Arginine vasopressin infusion significantly depressed the myocardial function in an ischemia/reperfusion model and increased mortality in comparison with both saline and dobutamine treated animals. The use of vasopressin may be contraindicated in non-vasodilatory shock states associated with significant cardiac injury. PMID- 19549334 TI - Akt finds its new path to regulate cell cycle through modulating Skp2 activity and its destruction by APC/Cdh1. AB - Skp2 over-expression has been observed in many human cancers. However, the mechanisms underlying elevated Skp2 expression have remained elusive. We recently reported that Akt1, but not Akt2, directly controls Skp2 stability by interfering with its association with APC/Cdh1. As a result, Skp2 degradation is protected in cancer cells with elevated Akt activity. This finding expands our knowledge of how specific kinase cascades influence proteolysis governed by APC/Cdh1 complexes. However, it awaits further investigation to elucidate whether the PI3K/Akt circuit affects other APC/Cdh1 substrates. Our results further strengthen the argument that different Akt isoforms might have distinct, even opposing functions in the regulation of cell growth or migration. In addition, we noticed that Ser72 is localized in a putative Nuclear Localization Sequence (NLS), and that phosphorylation of Ser72 disrupts the NLS and thus promotes Skp2 cytoplasmic translocation. This finding links elevated Akt activity with the observed cytoplasmic Skp2 staining in aggressive breast and prostate cancer patients. Furthermore, it provides the rationale for the development of specific Akt1 inhibitors as efficient anti-cancer therapeutic agents. PMID- 19549335 TI - Filtering genes for cluster and network analysis. AB - BACKGROUND: Prior to cluster analysis or genetic network analysis it is customary to filter, or remove genes considered to be irrelevant from the set of genes to be analyzed. Often genes whose variation across samples is less than an arbitrary threshold value are deleted. This can improve interpretability and reduce bias. RESULTS: This paper introduces modular models for representing network structure in order to study the relative effects of different filtering methods. We show that cluster analysis and principal components are strongly affected by filtering. Filtering methods intended specifically for cluster and network analysis are introduced and compared by simulating modular networks with known statistical properties. To study more realistic situations, we analyze simulated "real" data based on well-characterized E. coli and S. cerevisiae regulatory networks. CONCLUSION: The methods introduced apply very generally, to any similarity matrix describing gene expression. One of the proposed methods, SUMCOV, performed well for all models simulated. PMID- 19549336 TI - Palliative care for older people - exploring the views of doctors and nurses from different fields in Germany. AB - BACKGROUND: Providing appropriate palliative care for older people is a major task for health care systems worldwide, and up to now it has also been one of the most neglected. Focusing on the German health care system, we sought to explore the attitudes of health professionals regarding their understanding of palliative care for older patients and its implementation. METHODS: In a qualitative study design, focus groups were established consisting of general practitioners, geriatricians, palliative care physicians, palliative care nurses and general nurses (a total of 29 participants). The group discussions were recorded, transcribed, coded and analysed using the methodological approach of Qualitative Description. RESULTS: Deficiencies in teamwork and conflicting role definitions between doctors and nurses and between family practitioners and medical specialists were found to be central problems affecting the provision of appropriate palliative care for older people. It was emphasized that there are great advantages to family doctors playing a leading role, as they usually have the longest contacts to the patients. However, the professional qualifications of family doctors were to some extent criticized. The general practitioners for their part criticized the increasing specialization on the field of palliative care. All groups complained that the German compensation system gives insufficient consideration to the time-consuming care of older patients, and about excessive bureaucracy. CONCLUSION: General practitioners are the central health professionals in the delivery of palliative care for older people. They should however be encouraged to involve specialized services such as palliative care teams where necessary. With the German health care reform of 2007, a legal framework has been created that allows for this. As far as its realization is concerned, it must be ensured that the spotlight remains on the needs of the patients and not on policy conflicts and rivalries between health care professionals. Older people might particularly benefit if "talking" medicine and time-consuming care were properly catered for, financially and organizationally, in the health care system. PMID- 19549337 TI - Effects of cIAP-1, cIAP-2 and XIAP triple knockdown on prostate cancer cell susceptibility to apoptosis, cell survival and proliferation. AB - BACKGROUND: Manipulating apoptotic resistance represents an important strategy for the treatment of hormone refractory prostate cancer. We hypothesised that the Inhibitor of Apoptosis (IAP) Proteins may be mediating this resistance and knockdown of cIAP-1, cIAP-2 and XIAP would increase sensitivity to apoptosis. METHODS: cIAP-1, cIAP-2 and XIAP where knocked down either individually or in combination using siRNA in androgen independent prostate cancer PC-3 cells as confirmed by real-time PCR and western blotting. Cells were then treated with TRAIL, Etoposide, or Tunicamycin, and apoptosis assessed by PI DNA staining. Apoptosis was confirmed with Annexin V labelling and measurement of PARP cleavage, and was inhibited using the pan-caspase inhibitor, zVAD.fmk. Clonogenic assays and assessment of ID-1 expression by western blotting were used to measure recovery and proliferation. RESULTS: PC-3 are resistant to TRAIL induced apoptosis and have elevated expression of cIAP-1, cIAP-2 and XIAP. Combined knockdown sensitised PC-3 to TRAIL induced apoptosis, but not to Etoposide or Tunicmycin, with corresponding increases in caspase activity and PARP cleavage which was inhibited by ZVAD.fmk. Triple knock down decreased proliferation which was confirmed by decreased ID-1 expression. CONCLUSION: Simultaneous knock down of the IAPs not only sensitised the PC-3 to TRAIL but also inhibited their proliferation rates and clonogenic survival. The inability to alter sensitivity to other triggers of apoptosis suggests that this effect is specific for death receptor pathways and knock down might facilitate immune-surveillance mechanisms to counter cancer progression and, in combination with therapeutic approaches using TRAIL, could represent an important treatment strategy. PMID- 19549338 TI - A pneumatically powered knee-ankle-foot orthosis (KAFO) with myoelectric activation and inhibition. AB - BACKGROUND: The goal of this study was to test the mechanical performance of a prototype knee-ankle-foot orthosis (KAFO) powered by artificial pneumatic muscles during human walking. We had previously built a powered ankle-foot orthosis (AFO) and used it effectively in studies on human motor adaptation, locomotion energetics, and gait rehabilitation. Extending the previous AFO to a KAFO presented additional challenges related to the force-length properties of the artificial pneumatic muscles and the presence of multiple antagonistic artificial pneumatic muscle pairs. METHODS: Three healthy males were fitted with custom KAFOs equipped with artificial pneumatic muscles to power ankle plantar flexion/dorsiflexion and knee extension/flexion. Subjects walked over ground at 1.25 m/s under four conditions without extensive practice: 1) without wearing the orthosis, 2) wearing the orthosis with artificial muscles turned off, 3) wearing the orthosis activated under direct proportional myoelectric control, and 4) wearing the orthosis activated under proportional myoelectric control with flexor inhibition produced by leg extensor muscle activation. We collected joint kinematics, ground reaction forces, electromyography, and orthosis kinetics. RESULTS: The KAFO produced approximately 22%-33% of the peak knee flexor moment, approximately 15%-33% of the peak extensor moment, approximately 42%-46% of the peak plantar flexor moment, and approximately 83%-129% of the peak dorsiflexor moment during normal walking. With flexor inhibition produced by leg extensor muscle activation, ankle (Pearson r-value = 0.74 +/- 0.04) and knee ( r = 0.95 +/ 0.04) joint kinematic profiles were more similar to the without orthosis condition compared to when there was no flexor inhibition (r = 0.49 +/- 0.13 for ankle, p = 0.05, and r = 0.90 +/- 0.03 for knee, p = 0.17). CONCLUSION: The proportional myoelectric control with flexor inhibition allowed for a more normal gait than direct proportional myoelectric control. The current orthosis design provided knee torques smaller than the ankle torques due to the trade-off in torque and range of motion that occurs with artificial pneumatic muscles. Future KAFO designs could incorporate cams, gears, or different actuators to transmit greater torque to the knee. PMID- 19549339 TI - Comparative genomics and evolution of conserved noncoding elements (CNE) in rainbow trout. AB - BACKGROUND: Recent advances in the accumulation of genetic mapping and DNA sequence information from several salmonid species support the long standing view of an autopolyploid origin of these fishes (i.e., 4R). However, the paralogy relationships of the chromosomal segments descendent from earlier polyploidization events (i.e., 2R/3R) largely remain unknown, mainly due to an unbalanced pseudogenization of paralogous genes that were once resident on the ancient duplicated segments. Inter-specific conserved noncoding elements (CNE) might hold the key in identifying these regions, if they are associated with arrays of genes that have been highly conserved in syntenic blocks through evolution. To test this hypothesis, we investigated the chromosomal positions of subset of CNE in the rainbow trout genome using a comparative genomic framework. RESULTS: Through a genome wide analysis, we selected 41 pairs of adjacent CNE located on various chromosomes in zebrafish and obtained their intervening, less conserved, sequence information from rainbow trout. We identified 56 distinct fragments corresponding to about 150 Kbp of sequence data that were localized to 67 different chromosomal regions in the rainbow trout genome. The genomic positions of many duplicated CNE provided additional support for some previously suggested homeologies in this species. Additionally, we now propose 40 new potential paralogous affinities by analyzing the variation in the segregation patterns of some multi-copy CNE along with the synteny association comparison using several model vertebrates. Some of these regions appear to carry signatures of the 1R, 2R or 3R duplications. A subset of these CNE markers also demonstrated high utility in identifying homologous chromosomal segments in the genomes of Atlantic salmon and Arctic charr. CONCLUSION: CNE seem to be more efficacious than coding sequences in providing insights into the ancient paralogous affinities within the vertebrate genomes. Such a feature makes these elements extremely attractive for comparative genomics studies, as they can be treated as 'anchor' markers to investigate the association of distally located candidate genes on the homologous genomic segments of closely or distantly related organisms. PMID- 19549341 TI - Medical decision making for patients with Parkinson disease under Average Cost Criterion. AB - Parkinson's disease (PD) is one of the most common disabling neurological disorders and results in substantial burden for patients, their families and the as a whole society in terms of increased health resource use and poor quality of life. For all stages of PD, medication therapy is the preferred medical treatment. The failure of medical regimes to prevent disease progression and to prevent long-term side effects has led to a resurgence of interest in surgical procedures. Partially observable Markov decision models (POMDPs) are a powerful and appropriate technique for decision making. In this paper we applied the model of POMDP's as a supportive tool to clinical decisions for the treatment of patients with Parkinson's disease. The aim of the model was to determine the critical threshold level to perform the surgery in order to minimize the total lifetime costs over a patient's lifetime (where the costs incorporate duration of life, quality of life, and monetary units). Under some reasonable conditions reflecting the practical meaning of the deterioration and based on the various diagnostic observations we find an optimal average cost policy for patients with PD with three deterioration levels. PMID- 19549340 TI - Monocytes and neutrophils expressing myeloperoxidase occur in fibrous caps and thrombi in unstable coronary plaques. AB - BACKGROUND: Myeloperoxidase (MPO) -containing macrophages and neutrophils have been described at sites of plaque rupture. The presence of these cells in precursor lesions to acute rupture (thin cap atheroma, or vulnerable plaque) and within thrombi adjacent to ruptures has not been described, nor an association with iron-containing macrophages within unstable plaques. METHODS: We studied 61 acute ruptures, 15 organizing ruptures, 31 thin cap fibroatheromas, and 28 fibroatheromas from 72 sudden coronary death victims by immunohistochemical and histochemical techniques. Inflammatory cells were typed with anti-CD68 (macrophages), anti-BP-30 (neutrophil bactericidal glycoprotein), and anti-MPO. Iron was localized by Mallory's Prussian blue stain. In selected plaques alpha smooth muscle actin (DAKO, Carpinteria, CA, clone M0851) was performed. RESULTS: MPO positive cells were present in 79% of ruptured caps, 28% of thin cap fibroatheroma, and no fibroatheromas; neutrophils were present in 72% of ruptures, 8% of thin cap fibroatheromas, and no fibroatheromas. Iron containing foam cells were present in the caps of 93% of acute ruptures, of 85% of organizing ruptures, 20% of thin cap atheromas, and 10% of fibroatheromas. MPO positive cells were more frequent in occlusive than non-occlusive thrombi adjacent to ruptures (p = .006) and were more numerous in diabetics compared to non-diabetics (p = .002) CONCLUSION: Unstable fibrous caps are more likely to contain MPO-positive cells, neutrophils, and iron-containing macrophages than fibrous caps of stable fibroatheromas. MPO-positive cells in thrombi adjacent to disrupted plaques are associated with occlusive thrombi and are more numerous in diabetic patients. PMID- 19549342 TI - Medication diaries do not improve outcomes with highly active antiretroviral therapy in Kenyan children: a randomized clinical trial. AB - BACKGROUND: As highly active antiretroviral therapy (HAART) becomes increasingly available to African children, it is important to evaluate simple and feasible methods of improving adherence in order to maximize benefits of therapy. METHODS: HIV-1-infected children initiating World Health Organization non-nucleoside reverse transcriptase-inhibitor-containing first-line HAART regimens were randomized to use medication diaries plus counselling, or counselling only (the control arm of the study). The diaries were completed daily by caregivers of children randomized to the diary and counselling arm for nine months. HIV-1 RNA, CD4+ T cell count, and z-scores for weight-for-age, height-for-age and weight-for height were measured at a baseline and every three to six months. Self-reported adherence was assessed by questionnaires for nine months. RESULTS: Ninety HIV-1 infected children initiated HAART, and were followed for a median of 15 months (interquartile range: 2-21). Mean CD4 percentage was 17.2% in the diary arm versus 16.3% in the control arm at six months (p = 0.92), and 17.6% versus 18.9% at 15 months (p = 0.36). Virologic response with HIV-1 RNA of <100 copies/ml at nine months was similar between the two arms (50% for the diary arm and 36% for the control, p = 0.83). The weight-for-age, height-for-age and weight-for-height at three, nine and 15 months after HAART initiation were similar between arms. A trend towards lower self-reported adherence was observed in the diary versus the control arm (85% versus 92%, p = 0.08). CONCLUSION: Medication diaries did not improve clinical and virologic response to HAART over a 15-month period. Children had good adherence and clinical response without additional interventions. This suggests that paediatric HAART with conventional counselling can be a successful approach. Further studies on targeted approaches for non-adherent children will be important. PMID- 19549343 TI - IGF-I and IGFBP-3 and the risk of lung cancer: a meta-analysis based on nested case-control studies. AB - BACKGROUND: Lung cancer is the leading cause of death from cancer worldwide. Conventional studies mainly think that insulin-like growth factor-I (IGF-I) and IGF-binding protein-3 (IGFBP-3) may promote and inhibit tumor growth, respectively. However, there are many different results about their function in some recent epidemiological studies. To evaluate the relationship between circulating serum levels of IGF-I, IGFBP-3 and lung cancer, a systematic review and meta-analysis of the published data was performed. METHODS: Literatures searched on PubMed and Embase databases were enrolled in the Meta-analysis. The Meta-analysis of all eligible studies was applied with Stata 10.0 software, and the pooled odds ratio(OR) and weighted mean difference (WMD) value were obtained. The Q test, Egger's test and Begg's funnel plot were used to evaluate the heterogeneity and publication bias between the studies. RESULTS: There are no statistically significant heterogeneity and publication bias between the studies. For IGF- I, the pooled OR and WMD were 0.87(95%CI: 0.60 approximately 1.13,) and 3.04(95%CI: -7.10 approximately 1.02, P = 0.14), respectively. For IGFBP-3, the pooled OR and WMD were 0.68(95%CI: 0.48 approximately 0.88,) and -112.28(95%CI: 165.88 approximately -58.68, P < 0.0001), respectively. CONCLUSION: The association between circulating IGF- I levels and the risk of lung cancer were not statistically significant; IGFBP-3, acts as a tumor suppressor and has a inverse correlation with the risk of lung cancer. PMID- 19549344 TI - Biochemical characterization of malate synthase G of P. aeruginosa. AB - BACKGROUND: Malate synthase catalyzes the second step of the glyoxylate bypass, the condensation of acetyl coenzyme A and glyoxylate to form malate and coenzyme A (CoA). In several microorganisms, the glyoxylate bypass is of general importance to microbial pathogenesis. The predicted malate synthase G of Pseudomonas aeruginosa has also been implicated in virulence of this opportunistic pathogen. RESULTS: Here, we report the verification of the malate synthase activity of this predicted protein and its recombinant production in E. coli, purification and biochemical characterization. The malate synthase G of P. aeruginosa PAO1 has a temperature and pH optimum of 37.5 degrees C and 8.5, respectively. Although displaying normal thermal stability, the enzyme was stable up to incubation at pH 11. The following kinetic parameters of P. aeruginosa PAO1 malate synthase G were obtained: Km glyoxylate (70 microM), Km acetyl CoA (12 microM) and Vmax (16.5 micromol/minutes/mg enzyme). In addition, deletion of the corresponding gene showed that it is a prerequisite for growth on acetate as sole carbon source. CONCLUSION: The implication of the glyoxylate bypass in the pathology of various microorganisms makes malate synthase G an attractive new target for antibacterial therapy. The purification procedure and biochemical characterization assist in the development of antibacterial components directed against this target in P. aeruginosa. PMID- 19549345 TI - Differential in vitro effects of insulin on Taenia crassiceps and Taenia solium cysticerci. AB - Hormones play a significant role in murine cysticercosis (Taenia crassiceps), and increase the frequency of porcine cysticercosis caused by Taenia solium. In the present study, we report the in vitro effect of insulin on the larval stages of T. crassiceps (ORF strain) and T. solium. In vitro exposure of T. crassiceps cysticerci to insulin was found to stimulate this parasite's reproduction twofold with respect to control values, while the same treatment had no effect on T. solium cysticerci. Moreover, normal female mice (BALB/cAnN) infected with T. crassiceps cysticerci previously exposed to insulin presented larger parasite loads than mice inoculated with vehicle-treated cysticerci. To determine the possible molecular mechanisms by which insulin affects T. crassiceps, the insulin receptor was amplified by means of reverse transcriptase-polymerase chain reaction (RT-PCR). Interestingly, both T. crassiceps and T. solium expressed the insulin receptor, although insulin had effects only on T. crassiceps. These results demonstrate that insulin has a dichotomistic effect; it acts directly only on T. crassiceps cysticerci reproduction, possibly through its binding to a specific insulin receptor synthesized by the parasite. Thus, insulin may be recognized by T. crassiceps cysticercus cells as a mitogenic factor, and contribute to parasite proliferation inside the host, as well as to the female mouse susceptibility to T.crassiceps. This phenomenon has not been reported for cysticercosis caused by T. solium, which could, in part, be related to the poor effect of insulin upon the human parasite. PMID- 19549346 TI - Hot topics in parenteral nutrition. A review of the use of glutamine supplementation in the nutritional support of patients undergoing bone-marrow transplantation and traditional cancer therapy. AB - The relationship between glutamine and malignancy can be traced back to the 1950s and the requirement for glutamine for malignant-cell growth in culture. Later studies demonstrated an association between the rate of proliferation of the malignant cells and glutamine usage. The excessive use of glutamine by malignant cells was seen as an opportunity for the development of a treatment using glutamine analogues, but unfortunately excessive toxicity was observed during clinical studies. In animal models glutamine supplementation, initially thought to increase tumour growth, actually causes tumour regression as a result of improved immune clearance of the tumour and appears to reduce the severity of the side effects of chemo- and radiotherapy. This finding led to human studies in both traditional cancer therapy and bone-marrow transplantation, which are reviewed here. Unfortunately, the majority of the studies performed are small and have poor methodological reporting. There is clinical heterogeneity in terms of routes of administration, dosing schedules, chemotherapy regimens and diseases. Studies of glutamine in non-bone-marrow transplantation chemo- and/or radiotherapy treatment suggest a possible trend towards reductions in objective mucositis but no effect on subjective symptoms. There is no evidence for its effect on other clinical outcomes. For bone-marrow transplantation there appears to be some benefit from oral glutamine in reducing mucositis and graft v. host disease, while intravenous glutamine may reduce infections but at the expense of an increased relapse rate. Good-quality studies are required in this area. PMID- 19549347 TI - Characterization of a leucine aminopeptidase of Babesia gibsoni. AB - Peptidases of parasitic protozoa are currently under intense investigation in order to identify novel virulence factors, drug targets, and vaccine candidates, except in Babesia. Leucine aminopeptidases in protozoa, such as Plasmodium and Leishmania, have been identified to be involved in free amino acid regulation. We report here the molecular and enzymatic characterization, as well as the localization of a leucine aminopeptidase, a member of the M17 cytosolic aminopeptidase family, from B. gibsoni (BgLAP). A functional recombinant BgLAP (rBgLAP) expressed in Escherichia coli efficiently hydrolysed synthetic substrates for aminopeptidase, a leucine substrate. Enzyme activity of the rBgLAP was found to be optimum at pH 8.0 and at 37 degrees C. The substrate profile was slightly different from its homologue in P. falciprum. The activity was also strongly dependent on metal divalent cations, and was inhibited by bestatin, which is a specific inhibitor for metalloprotease. These results indicated that BgLAP played an important role in free amino acid regulation. PMID- 19549348 TI - Vertical transmission of Toxoplasma gondii in Australian marsupials. AB - To date, little is known about the dynamics of vertical transmission of Toxoplasma gondii in Australian marsupials. Studies in mice demonstrate that vertical transmission of T. gondii is common and that chronically infected mice can transmit T. gondii to successive generations. In this study, PCR and immunohistochemistry were used to detect T. gondii in chronically infected marsupial dams and their offspring. T. gondii was detected in the unfurred pouch young of 2 out of 10 chronically infected western grey kangaroos (Macropus fuliginosus) and in the unfurred pouch young of a brush-tailed bettong (Bettongia penicillata). Results of the study suggest that vertical transmission of T. gondii can occur in chronically infected Australian marsupials. PMID- 19549350 TI - Tracking a heterosis effect in the field: tadpole resistance to parasites in the water frog hybridogenetic complex. AB - Depending on the extent of evolutionary divergence among parent taxa, hybrids may suffer from a breakdown of co-adapted genes or may conversely exhibit vigour due to the heterosis effect, which confers advantages to increased genetic diversity. That last mechanism could explain the success of hybrids when hybridization zones are large and long lasting, such as in the water frog hybridization complex. In this hybridogenetic system, hybrid individuals exhibit full heterozygosity that makes it possible to investigate in situ the impact of hybridization. We have compared parasite intensity between hybrid Rana esculenta and parental R. lessonae individuals at the tadpole stage in two populations inhabiting contrasted habitats. We estimated intensity of Gyrinicola sp. (Nematoda) in the gut, Echinostome metacercariae in the kidneys and Haplometra cylindracea in the body cavity (both species belong to Trematoda). Despite high sampling effort, no variation in parasite intensity was detected between taxa, except a possible higher tolerance to H. cylindracea in hybrid tadpoles. The low effect of hybridization suggests efficient gene co-adaptation between the two genomes that could result from hemiclonal selection. Variation in infection intensity among ponds could support the Red Queen hypothesis. PMID- 19549349 TI - Identification of the agent causing visceral leishmaniasis in Uzbeki and Tajiki foci by analysing parasite DNA extracted from patients' Giemsa-stained tissue preparations. AB - Our present study is the first attempt to characterize Leishmania parasites from foci in Uzbekistan and Tajikistan endemic for visceral leishmaniasis (VL). PCR sequencing of the ribosomal internal transcribed spacer 1 and multilocus microsatellite typing (MLMT) were applied to DNA extracted from preparations of Giemsa-stained bone marrow aspirates from 13 cases of VL. L. infantum was shown to cause VL currently occurring in this area. MLMT applying 14 microsatellite markers, previously shown to be polymorphic for strains of the L. donovani complex, revealed that microsatellite profiles of parasites causing human VL in the Namangan and Jizzakh regions in Uzbekistan, and Penjikent region in Tajikistan, basically coincide with those of strains of L. infantum MON-1. Furthermore, these parasites were assigned to a distinct cluster genetically clearly separated from the populations of L. infantum MON-1 from Europe, the Middle East and North Africa. The existence of a genetically homogeneous but distinct group of L. infantum MON-1 indicates that the parasites circulating in the Uzbeki and Tajiki foci studied have been restricted there for a long time rather than having been recently introduced from elsewhere by human or animal reservoir migration. PMID- 19549351 TI - A note on the editors of Parasitology. PMID- 19549352 TI - X-cell parasites in the European dab Limanda limanda are related to other X-cell organisms: a discussion on the potential identity of this new group of parasites. AB - Unusual tumour-like pathologies caused by mysterious cells termed 'X-cells' have been reported from numerous fish groups worldwide. After nearly 100 years of research, the tumour-like growths have recently been shown to be caused by a protozoan parasite. In the present study, histopathology and small subunit ribosomal DNA (SSU rDNA) sequences are used to assess whether the X-cell parasite infecting Atlantic dab Limanda limanda L. is distinct from the X-cell parasite infecting Japanese flounder and goby, and to determine their systematic position within the protists. SSU rDNA from Scottish dab was 89.3% and 86.7% similar to Japanese X-cell sequences from flounder and goby respectively, indicating that the parasite infecting dab in the Atlantic is distinct from the Pacific species. Histological studies revealed significant gill pathology and demonstrated the precise location of the parasites within the gill tissues using specific in situ hybridization probes. Phylogenetic analyses showed that the X-cell parasites from Scotland and Japan form a monophyletic group within the Myzozoa, and are basal alveolates. However, ultrastructure of X-cells from dab fails to confirm this systematic placement. PMID- 19549353 TI - Flow cytometric assessment of distinct physiological stages within Cryptosporidium parvum sporozoites post-excystation. AB - Cryptosporidium parvum are protozoan parasites responsible for outbreaks of gastrointestinal disease worldwide. Within the apical complex of this organism reside numerous vesicular secretory organelles and their discharge has been identified as essential for sporozoite motility, cell attachment and penetration. Traditionally, investigation of apical organelle discharge has relied on microscopic and immunochemical hybridization techniques. In this study we demonstrate for the first time how flow cytometry, in combination with vital dye staining, provides an avenue for discrimination of distinct physiological events occurring within Cryptosporidium sporozoites post-excystation. Time-course studies of freshly excysted sporozoites were carried out at 37 degrees C in cell free medium, stained with the fluorescent dyes SYTO9/PI, DiBAC4(3), Fluo-4 AM or FM1-43 and analysed by flow cytometry. Significant decreases in sporozoite plasma membrane permeability and increased membrane depolarization were found to be accompanied by concomitant increases in intracellular calcium. Subsequent to these changes, large increases in exocytosed vesicular membrane were apparent. In addition, by measuring side and forward angle light scatter we were able to assess changes in internal granularity and size of sporozoites post-excystation. These observations were suggestive of rapid mobilization, utilization and discharge of apical organelles within sporozoites, which we relate to changes in sporozoite infectivity, ATP levels and total secreted soluble protein. PMID- 19549354 TI - Using length-frequency data to elucidate the population dynamics of Argulus foliaceus (Crustacea: Branchiura). AB - This study uses a novel method for discriminating cohorts and investigating the population dynamics of the parasitic crustacean, Argulus foliaceus. Analysis of parasite length-frequency data was carried out in order to elucidate the timings and drivers behind the parasite's life cycle. Up to 6 cohorts of the parasite emerge through the course of 1 year in still-water trout fisheries in England. Recruitment ceases over the winter months; however, 3 cohorts of the parasite over-winter, 2 as eggs and 1 as a hatched stage. The technique, when used in conjunction with temperature data, also allowed for the reliable prediction of growth rates and provided estimates of egg incubation times and the length of hatching periods. These data showed that growth rates increased exponentially between the observed temperatures of 4 to 22 degrees C. The method allowed for the time taken from hatching to egg laying under field conditions to be predicted and produced estimates that were validated against independent laboratory studies on the growth of the parasite. PMID- 19549355 TI - P-glycoprotein interfering agents potentiate ivermectin susceptibility in ivermectin sensitive and resistant isolates of Teladorsagia circumcincta and Haemonchus contortus. AB - P-glycoprotein (P-gp) homologues, belonging to the ATP Binding Cassette (ABC) transporter family, are thought to play an important role in the resistance of gastro-intestinal nematode parasites against macrocyclic lactones. The aim of this study was to investigate the influence of various P-gp interfering compounds on the efficacy of ivermectin (IVM) in sensitive and resistant nematode isolates. The feeding of IVM resistant and sensitive Teladorsagia circumcincta and Haemonchus contortus first-stage larvae (L1) was assessed using a range of IVM concentrations (0.08-40 nm) with or without P-gp inhibitors: valspodar, verapamil, quercetin, ketoconazole and pluronic P85. The P-gp inhibitors were selected on the basis of their ability to interfere with P-gp transport activity in an epithelial cell line over-expressing murine P-gp. In the presence of P-gp interfering agents, the in vitro susceptibility to IVM of both sensitive and resistant isolates of T. circumcincta and H. contortus was increased. These results show that compounds interfering with P-gp transport activity could enhance IVM efficacy in sensitive isolates, and also restore IVM sensitivity in resistant nematodes. These results support the view that ABC transporters can play an important role in resistance to IVM, at least in the free-living stages of these economically important gastro-intestinal nematodes. PMID- 19549356 TI - [Relationship between NQO1C(609T), RAD51(G135C), XRCC3(C241T) single nucleotide polymorphisms and acute lymphoblastic leukemia]. AB - This study was purposed to investigate the relationship between NQO1C(609T), RAD51(G135C), XRCC3(C241T) single nucleotide polymorphisms and incidence of acute lymphoblastic leukemia (ALL). NQO1C(609T), RAD51(G135C), XRCC3(C241T) genotypes were detected by PCR-RFLP in 170 patients with de novo ALL and 458 normal persons as control. The results indicated that the genotype ratio of NQO1C(609T), RAD51(G135C) and XRCC3(C241T) in single genotype analysis showed no statistical difference between ALL patients and normal controls, which suggested that the single genotype affect onset of ALL without statistical significance. In combined genotype analysis, presence of both variants for NQO1C(609T) and RAD51(G135C) increased onset risk of ALL with myeloid antigen positive and with balanced translocation (OR value 5.553 and 2.618 respectively); the presence of homozygosity variant for NQO1C(609T) increased onset risk of ALL in the country children (OR = 2.541). In conclusion, the combined effect of NQO1C(609T), RAD51(G135C) and XRCC3(C241T) genotypes may promote occurrence of ALL, which suggests that the combined analysis of 3 genotypes has more predictive significance for ALL than single genotype analysis. PMID- 19549357 TI - [Expression of annexin II in patients with hematologic malignancies]. AB - This study was purposed to investigate the expression of annexin II in patients with hematologic malignancies and its role in genesis and development of hematologic malignancies. The expression levels of annexin II in bone marrow cells from untreated 81 patients with acute leukemia, 6 patients with MM and 20 patients with iron deficiency anemia were detected by real-time PCR assay. The results showed that the expression of annexin II mRNA significantly increased in M(3) patients as compared with others, the expression of annexin II gene in groups M(5), MM, M(4) were higher than that of other groups except M(3) group, and there were no significant difference in expression of annexin II gene between M(1) + M(2) groups and controls. It is concluded that the expression of annexin II gene significantly increased in patients M(5), M(4), MM, who showed higher ratio of infiltration than other patients. It is inferred that the annexin II participates in invasion and infiltration of hematologic malignancies probably through enhancing the degradation of extracellular matrix by cells of hematologic malignancies. PMID- 19549358 TI - [Hypoxia and its simulant CoCl(2) down-regulates Foxp3 expression independent from HIF-1alpha]. AB - This study was purposed to investigate the expression of Jurkat cell Foxp3 in hypoxia condition and the role of HIF-1alpha in this process as well as to clarify the mechanism influencing function of regulatory T cells by hypoxia. The Jurkat cells were incubated with hypoxia (1% O(>2)) and its simulant CoCl(2) for different times (0, 6, 12, 24 hours), the viability was measured by trypan blue staining, the expression of HIF-1alpha was detected by Western blot, the expression of Foxp3 was detected by real-time PCR, the expressions of HIF-1alpha and Foxp3 were assayed after HIF-1alpha in Jurkat cells was inhibited by using RNA interference technique. The results indicated that after Jurkat cells were treated with hypoxia and its simulant CoCl(2), the significant accumulation of HIF-1alpha in cells appeared, but the expression of Foxp3 was obviously down regulated; after expression of HIF-1alpha in Jurkat cells was inhibited by siRNA interference, the CoCl(2) still could down-regulate the expression of Foxp3. It is concluded that the hypoxia and its simulant CoCl(2) can obviously down regulate the expression of Foxp3, but this process is independent from HIF 1alpha. PMID- 19549360 TI - [Tyrosine kinase JAK2V617F mutation in human myeloproliferative disorders]. AB - This study was aimed to investigate the JAK2V617F mutation in myeloproliferative disorders (MPD) and to evaluate the significance of JAK2V617F in diagnosis and therapy of MPD. The bcr/abl fusion gene in 70 MPD patients was detected by reverse transcription polymerase chain reaction (PT-PCR). The JAK2V617F mutation was detected by allele-specific polymerase chain reaction (AS-PCR) and the results were confirmed by sequence analysis. The results indicated that the bcr/abl fusion gene could be detected in 38 patients with chronic myeloid leukemia (CML), but not in the 32 none-CML patients. The JAK2V617F mutation was detected in 12 out of 16 (75%) patients with polycythemia vera (PV), 3 out of 10 (30%) patients with essential thrombocythemia (ET), 3 out of 6 (50%) patients with idiopathic myelofibrosis (IMF), but not in any of the CML patients. The JAK2V617F mutation frequencies between CML and bcr/abl negative MPD patients were statistically significant (p < 0.05). It is concluded that the JAK2V617F may be a characteristic molecular event in PV, ET and IMF patients which may serve as an important molecular marker for the diagnosis and classification of the three diseases. PMID- 19549359 TI - [Analysis of variant translocation der ins (17; 15) in patient with APL by G banding technique and interphase fluorescence in situ hybridization]. AB - To investigate the biological characteristics of the variant translocation der ins (17;15) in a patient with acute promyelocytic leukemia (APL), the conventional G-banding technique, interphase fluorescence in situ hybridization (int-FISH), RT-PCR, gene scanning, gene sequence and flow cytometry were performed. The results indicated that the variant translocation der ins (17, 15) observed by G banding technique was a rare type, the int-FISH assay by using dual color pml/raralpha fusion probes confirmed the cytogenetic findings. The detection results of other molecular methods demonstrated the existence of the whole pml/raralpha fusion gene, while this case had insertion variant translocation. This patient got complete remission by using combined chemotherapy, and survives with continuous complete remission during following up for 1 year. In conclusion, the variant translocation der ins (17; 15) is rare type in APL, its incidence is lower, several signal types in detection of int FISH were observed and the combination chemotherapy for this patient showed more obvious efficacy. PMID- 19549361 TI - [Influences of bone marrow mesenchymal stem cells in patients with acute myeloid leukemia and non-leukemia on HL-60 cells -- a comparison study]. AB - This study was aimed to compare the influences of bone marrow mesenchymal stem cells (BMMSCs) from patients with acute myeloid leukemia (AML), AML patients with complete remission (CR) and non-leukemia patients on HL-60 cells. The HL-60 cells were divided into three groups: group of co-cultivation with BMMSCs of AML patients, group of co-cultivation with BMMSCs of AML patients with CR and group of co-cultivation with BMMSCs of non-leukemia patients. The count of HL-60 cells, the CD11b and survivin expression of HL-60 cells, the cell cycle distribution of the HL-60 cells in 3 groups were compared by flow cytometry, the morphology and differentiation rate of HL-60 cells in 3 groups were observed and compared by microscopy. The results showed that there were no differences in HL-60 cell count at five and seven days, in HL-60 distribution at the G(0)/G(1) phase, in survivin and CD 11b expressions in 3 groups. All cells of 3 groups began to mature, and the differentiation rates in 3 groups were 18.0 +/- 3, 17.0 +/- 1.3 and 19.0 +/- 2.0 respectively, therefore there were no significant differences between the 3 groups (p = 0.23). It is concluded that there is no influence of BMMSCs in 3 groups on the proliferation and differentiation of HL-60 cells. PMID- 19549362 TI - [Prognostic significance of minimal residual disease detected by multiparameter flow cytometry in acute myeloid leukemia]. AB - This study was aimed to explore prognostic significance of minimal residual disease (MRD) detection in patients with acute myeloid leukemia (AML) by multiparameter flow cytometry (MCF). Leukemia-associated immunophenotype (LAIP) of newly diagnosed AML patients were determined by 4-color 5 antibody panels and patients with sensitive LAIP were chosen for MRD detection. 601 bone marrow samples from 95 patients were acquired after treatment and MRD were considered positive by the critical normal value plus twice standard deviation in normal bone marrow specimen. The patients were divided into three groups and the clinical significance was analyzed every 2 months within initial 6 months after induction treatment. The results showed that the relapse rate and relapse-free survival (RFS) rate were all significantly different between MRD positive and MRD negative patients in the three groups (p < 0.05). Patients with MRD positive had a median relapse-free survival time of 11 months, 11.5 months and 11 months at 1 2, 3 - 4 and 5 - 6 months respectively, while all patients with MRD negative were not observed to reach median release-free survival time (p < 0.05). Furthermore, the clinical significance was analyzed after induction and one course of consolidate treatment, the relapse rate of MRD positive and MRD negative patients were 57.14% versus 0% and 91.67% versus 2.27% respectively (p = 0.000 and p = 0.000). It is concluded that MRD detection by multi-parameter flow cytometry can predict outcome of AML patients, which should be continuously monitored after treatment. PMID- 19549363 TI - [Prognosticating relapse risk based on multiparameter flow cytometric assessment of minimal residual disease in patients with acute myeloid leukemia]. AB - The objective of this study was to investigate the prognosticating value of multiparameter flow cytometry in detection of minimal residual disease (MRD) and relapse risk of patients with acute myeloid leukemia (AML). Multiparameter flow cytometry (MPFC) analysis was used to detect the leukemia-associated aberrant immunophenotype (LAIP) of the pretreated patients with AML and to assess the levels of MRD after remission induction (Post-Ind MRD) and consolidation therapy (Post-Cons MRD). The results showed that the definite LAIP could be detected in 94.3% of the patients (115/122) with AML (except APL). Among 115 cases only one LAIP was identified in 15 cases (13.0%), but two or more LAIP were identified in other 100 cases (87.0%). The most frequent LAIP identified was cross-lineage antigen expression (40.9%). The percentages of asynchronous antigen expression, antigen over-expression and antigen lack expression were 20.9%, 27.0%and 34.8% respectively. MRD frequency was monitored in 41 AML patients with CR after remission induction chemotherapy and 2 or more cycles of consolidation chemotherapy. 24 patients were Post-Ind MRD(+) and 17 patients were Post-Ind MRD( ). The percentages of relapse in cases of Post-Ind MRD(+) and Post-Ind MRD(-) were 75.0% (18/24) and 29.4% (5/17) respectively after consolidation chemotherapy. The relapse free survival (RFS) times of the patients with Post-Ind MRD(+) and Post-Ind MRD(-) were 49.06 +/- 6.53 months and 11.92 +/- 1.64 months (p < 0.0001) respectively. 18 patients were Post-Cons MRD(+) and 23 patients were Post-Cons MRD(-). The percentages of relapse in cases of Post-Cons MRD(+) and Post-Cons MRD(-) patients were 100% (18/18) and 21.7% (5/23) respectively after consolidation chemotherapy. The RFS times of the patients with Post-Cons MRD(+) and Post-Cons MRD(-) were 41.74 +/- 5.52 months and 10.06 +/- 1.72 months (p < 0.0001) respectively. It is concluded that the levels of post-Ind MRD and post Cons MRD identified in the patients with AML was highly associated with their RFS. The detection of MRD by MPFC provides prognostic information in AML patients. PMID- 19549364 TI - [Reversal of leukemia multidrug resistance by sequence-specific short hairpin RNA]. AB - This study was aimed to design and screen short hairpin RNA (shRNA) molecules targeting multidrug resistance gene (mdr1), as well as to investigate the effects of shRNA expression vector on K562/A02 cells. Mdr1-shRNA expression vector was transfected into K562/A02 cells by lipofectamine 2000, and G418 was added to screen and establish the stable expression cell strain. The expressions of mdr1 mRNA and protein were detected by real-time RT-PCR and Western blot respectively. The sensitivity of cells to chemodrugs after interference were tested by CCK8 assay. The function of p-glycoprotein was determined by Rhodamine 123 efflux experiment. The results showed that all of 4 mdr1-shRNA expression vectors could significantly knockdown the expression of p-glycoprotein as compared with control vector, moreover, the vector targeting 508 - 526 sites of mdr1 gene was the best one. It is concluded that the mdr1-shRNA expression vector gained by screening can significantly knockdown the expression of mdr1 gene and reverse leukemia drug resistance, paving the way for the application of RNAi in the following animal experiments. PMID- 19549365 TI - [Effect of intracellular acidification on P-glycoprotein in drug-resistant K562/A02 cells]. AB - The aim of this study was to investigate the effect of intracellular acidification on the P-gp in K562/A02 cells. Confocal laser microscope was used to determine the intracellular acidification. MTT assay was used to detect the cytotoxicity of intracellular acidification on K562 and K562/A02 cells. Flow cytometry was applied to measure the influence of intracellular acidification on the activity of P-gp. The P-gp expression at protein and mRNA levels were determined by Western blot and real-time RT-PCR respectively. The results indicated that intracellular acidification had no obvious cytotoxicity on K562 and K562/A02 cells. The function of P-gp in K562/A02 cells weakened along with decrease of intracellular acidification, the intracellular acidification significantly increased the accumulation of Rhodamine 123 (Rh 123) and suppressed the efflux of Rh 123 mediated by P-gp. The intracellular acidification also inhibited the expression of P-gp in K562/A02 cells at protein and mRNA levels which showed intracellular acidification with time-dependence. It is concluded that the intracellular acidification can inhibit the expression and function of P gp in K562/A02 cells. PMID- 19549366 TI - [Effect of proteasome inhibitors MG-132 at different doses on cultured K562 cell apoptosis]. AB - This study was aimed to investigate the effect of proteasome inhibitor MG-132 at different doses on cultured K562 cell apoptosis. MTT assay was used to observe the activity of K562 cell proliferation inhibition rate after treating for 48 hours at different doses (0, 2, 4, 8, 16, 32 micromol/L). Immunocytochemistry was used to detect the NF-kappaB activity and glucocorticoid receptor (GR) expression. Flow cytometry was used to determine the K562 cell apoptosis. The results indicated that proliferation inhibition rate of K562 cells after treated for 48 hours showed dose-dependent, the inhibitory rates of cell proliferation in test groups were significant higher than that in control group, and the effect in 32 micromol/L test group was the most obvious (45.24 +/- 4.12)% (p < 0.05). The NF-kappaB activity and GR expression after treating for 48 hours showed dose dependent. Compared with control group, the NF-kappaB activities in test groups were lower (p < 0.05), and the NF-kappaB activity in 32 micromol/L test group was the lowest (63.60 +/- 2.95); the GR expression in test groups was higher (p < 0.05), and the GR expression in 16 micromol/L test group was the highest (75.62 +/- 2.70). The K562 cell apoptosis rate after treating for 48 hours also showed dose-dependent. Compared with control group, the K562 cell apoptosis rates in test groups were higher (p < 0.05), the K562 cell apoptosis rate in 32 micromol/L test group was the highest (21.37 +/- 2.02)%. It is concluded that the MG-132 may induce K562 cell apoptosis and proliferation inhibition through up-regulation of NF-kappaB activity and down-regulation of GR expression both in dose-dependent manner. PMID- 19549367 TI - [Influence of As(2)O(3) on proteasome beta(1)-subunit in NB4 cells]. AB - This study was aimed to explore the correlation between effects of arsenic trioxide on NB4 cell differentiation and the change of beta(1)-subunit of 26S proteasome. NB4 cell in 0.5 micromol/L As(2)O(3) was incubated for 24 hours and 48 hours, then total protein was extracted, expressions of subunit beta(1) and PML-RARalpha fusion protein were determined by Western blot. The results indicated that the expression of 26S proteasome beta(1)-subunit increased after incubation with As(2)O(3) for 24 hours, but after culture with As(2)O(3) for 48 hours, the expression of beta-subunit decreased to the baseline. Meanwhile, the expression of PML-RARalpha fusion protein obviously decreased after 24 hours, and kept low level at 48 hours. It is concluded that the expression of 26S proteasome beta(1)-subunit increases after exposure to As(2)O(3). Increment of 26S proteasome beta(1)-subunit may be associated with the degradation of PML-RARalpha fusion protein and plays roles in the differentiation and apoptosis of NB4 cells. PMID- 19549368 TI - [Autologous regulatory T cells can suppress the proliferation of lymphoma cell line in vitro]. AB - This study was aimed to investigate the suppressive effect of regulatory T (Treg) cells on the T cell lymphoma EL4 cell line and to explore its mechanism. C57BL/6 Mouse Treg cells were isolated by MACS (magnetic cell sorting). The purity and the expression of Foxp3 were detected by flow cytometry. The suppressive effect of sorted Treg cells on EL4 cells was detected by MTT assay. The secretion of TGF beta1 and IL-10 was examined by enzyme-linked immunosorbent assay (ELISA). The results showed that CD4(+)CD25(+) T cells could be successfully isolated by MACS with the purity reaching 91.6% and the expression level of Foxp3 was 78.9%. The ratio of viable cells was more than 95%. Regulatory T cells could suppress the proliferation of EL4 cells effectively in the presence of antigen presenting cells (APCs). And the suppressive effect was most significant at 1:1 ratio. In addition, the suppression still existed without APCs. TGF-beta1 and IL-10 could not be detected by ELISA. It is concluded that the Treg cells can suppress T lymphoma cell in vitro. The suppressive effect of Treg cells works in dose dependent manner, but not in cytokine-dependent manner. The mechanism of this suppression may take effect through cell-cell contact. PMID- 19549369 TI - [Killing effect of ZnPcH(1)-PDT on lymphoma cells]. AB - The objective of this study was to investigate the effect of ZnPcH(1)-PDT on the lymphoma cells and its mechanism. Human Burkitt's lymphoma cell line CA46 and mouse lymphoma cell line P388 were selected as objects for study. The killing effect of ZnPcH(1)-PDT on cells were assessed by MTT method and colony formation assay; the cell death patterns were analyzed by AO/EB fluorescence stain, TdT mediated dUTP nick end labeling (TUNEL), DNA ladder assay; and the different proportions of each death pattern were determined by Annexin-V(-FITC)/PI double stains. The results showed that ZnPcH(1)-PDT displayed anti-proliferation effect on both CA46 cells and P388 cells in dose-dependent manner. CA46 cells were less sensitive to PDT than P388 cells (p < 0.05). Furthermore, PDT could induce cell apoptosis in time-dependent manner. The rate of cell apoptosis increased in the PDT-treated cells. The results of Annexin-V(-FITC)/PI stain indicated that early apoptosis was the main death pattern in the PDT-treated CA46 cells, while early apoptosis and necrosis were the main death model in the PDT-treated P388 cells. It is concluded that ZnPcH(1)-PDT can effectively inhibit lymphoma cell proliferation and induce cell apoptosis. PMID- 19549370 TI - [Apoptosis of Burkitt's lymphoma Raji cell line induced by bortezomib]. AB - The aim of this study was to clarify whether bortezomib might induce apoptosis in Burkitt's lymphoma Raji cell line and its mechanism. Different concentrations of bortezomib were used to treat Raji cells and its effects of time and dose were observed. Cell morphology was observed under light microscope; flow cytometry was used to analyze cell apoptosis; RT-PCR was used to detect the expressions of NF kappaB and p53 gene mRNAs. The results showed that the bortezomib could inhibit Raji cell growth within a certain range of treating time and dose. Apoptosis were induced in relation to time and dose. The expression of NF-kappaB mRNA and p53 mRNA decreased after treatment with bortezomib. It is concluded that the bortezomib can induce Raji cell apoptosis, which provides a theoretical basis for clinical treatment. NF-kappaB and p53 gene are supposed to participate in the bortezomib induced apoptosis of Raji cells. PMID- 19549371 TI - [Effect of arsenic trioxide on apoptosis in lymphoblastoid Raji cell line with relation to expression of mcl-1 gene]. AB - The aim of this study was to investigate the effect of different concentrations of arsenic trioxide (As(2)O(3)) on apoptosis of lymphoblastoid Raji cell line and its possible mechanisms, as well as its relation to the expression of mcl-1 gene. The Raji cells were treated with different concentrations of As(2)O(3), the classical DNA ladder of cell apoptosis was detected by agar gel electrophoresis, the changes of mitochondrial membrane potential in Raji cells were assayed by confocal laser scanning microscopy, the changes of mcl-1 gene expression after exposure of cells to As(2)O(3) was detected by real-time quantitative fluorescence PCR. The results indicated that 1 micro mol/L As(2)O(3) did not lead to significant apoptosis of Raji cells, 2 - 8 micromol/L As(2)O(3) induced Raji cell apoptosis. Along with increase of drug concentration, the mitochondrial respiratory function and membrane potential of Raji cells obviously decreased. At same time, the expression level of mcl-1 gene were significantly down-regulated. It is concluded that As(2)O(3) can markedly decrease mitochondrial respiratory function and membrane potential of Raji cells, and down-regulate expression of mcl-1 gene, which may be the mechanisms resulting in cell apoptosis. PMID- 19549372 TI - Preliminary study on 5-azacytidine anti-myeloma activity in vitro. AB - This study was aimed to investigate the effect of 5-azacytidine (5-AZA) on XAF1 expression in myeloma cells and efficacy of 5-AZA treatment for myeloma in vitro. XAF1 expression was analyzed by semi-quantitative PCR. Methylation-specific PCR (MSP) was used to detect the methylation status of XAF1 promoter CpG islands. RPMI 8226 and XG-7 cells were treated with 0-5 micromol/L of 5-AZA. Expression of XAF1 mRNA variants was confirmed by gel electrophoresis. The results indicated that the untreated RPMI 8226 cell expressed XAF1 mRNA transcript 1 and transcript 2, untreated XG-7 cells did not express XAF1 mRNA. Hypermethylation of XAF1 promoter CpG islands could be detected in both cell lines. Both cell lines expressed full-length XAF1 transcript after being treated with 2.5 micromol/L of 5-AZA for 72 hours. 5-AZA treatment led XAF1 promoter CpG island to hypomethylation in both cell lines. 5-AZA exerted anti-myeloma activity in a time and concentration-dependent manner. The IC(50) value of XG-7 cells treated with 5-AZA for 48 hours was 2.6 micromol/L. 1.0, 2.0, 2.5 and 5.0 micromol/L of 5-AZA treatment for 48 hours induced (34.3 +/- 8.0)%, (54.8 +/- 3.1)%, (64.1 +/- 3.4)%, (81.0 +/- 4.1)% apoptosis in XG-7 cell line respectively. The combination of 1.0 4.0 micromol/L of 5-AZA with 1.0 - 4.0 micromol/L of arsenic trioxide (ATO) exhibited synergistic toxicity in myeloma cells with all CI values less than 1.0. It is concluded that lack of XAF1 expression and abnormal expression of XAF1 in myeloma cell lines are associated with the hypermethylation of XAF1 gene promoter CpG island. 5-AZA treatment can induce the expression of XAF1 mRNA and protein in myeloma. 5-AZA exerts anti-myeloma activity via apoptosis at clinically achievable concentrations. The findings suggested that 5-AZA and ATO may be an effective combination in the therapy of patients with multiple myeloma. PMID- 19549373 TI - [Osteoclast differentiation regulated by receptor activator of nuclear factor kappaB probably through a novel signaling pathway]. AB - This study was aimed to investigate the signaling pathways regulating osteoclast (OC) differentiation by receptor activator of nuclear factor kappa (RANK) under physiological condition so as to provide some theoretical basis for clarifying mechanism of bone destruction in multiple myeloma. A mutant TNFR(1)/RANK(2) (named RANK-Mu) chimera consisting of tumor necrosis factor receptor 1 (TNFR(1)) and RANK intramembrane domain was constructed by using deletion mutation for deleting IVVY amino acids in RANK intramembrane domain in accordance with (535 )IVVY(-538) as specific domain regulating OC differentiation by RANK. The RANK-Mu and TNFR(1)/RANK chimera without mutation (RANK-WT) were packaged by using plat E cell line to produce the retrovirus, which were transfected into bone marrow macrophages (BMMs) of TNFR(1)/TNFR(2) double knockout mice. After stimulation of these transfected BMMs with TNF-alpha, the differentiation of BMMs into OCs were observed, meanwhile the phosphorylation of NF-kappab, JNK, p38 and ERK was detected by Western blot after stimulation of these BMMs with TNF-alpha. The results showed that BMMs transfected with RANK-WT could be differentiated into OCs and phosphorylation of NF-kappaB, JNK, P38 and ERK were activated at 5 - 10 minutes after being stimulated by TNFalpha. BMMs transfected with RANK-Mu could not be differentiated into OCs, but phosphorylation of NF-kappaB, JNK, P38 and ERK were activated also. It is concluded that RANK regulates osteoclast differentiation probably not through 4 typical signaling pathways, named as NF kappaB, JNK, P38 and ERK, in this process other new signaling pathways maybe participate. PMID- 19549374 TI - Protein RAP1GAP in human myelodysplastic syndrome detected by flow cytometry and its clinical relevance. AB - Previous study on the gene expression profile of human MDS by using microarray discovered that transcription of RAP1GAP was up-regulated, which was confirmed by quantitative RT-PCR in expanding cohort of MDS patients. This study was pourposed to investigate the expression of RAP1GAP in human MDS and its clinical relevance. The expression of RAP1GAP in bone marrow cells of 19 MDS patients was detected by flow cytometry and was compared with that in patients with non-malignant blood diseases and acute leukemias, meanwhile the relevance between expression level of RAP1GAP and hemoglobin, leukocytes, platelets, blasts percentage in bone marrow cells and IPSS score was analyzed. The results indicated that the expression level of RAP1GAp in MDS patients significantly increased as compared with patients with non-malignant blood diseases or AML (8.42 +/- 8.37% vs 2.97 +/- 4.75% or 2.26 +/- 4.24%). Among MDS patients, the expression level of RAP1GAP in MDS-RA was significantly higher than that in MDS-RAEB (11.64 +/- 9.07% vs 4.37 +/ 4.65%). However, no definitive correlation of expression level with above mentioned clinical parameters was found in detected patients with DMS. In conclusion, the expression of RAP1GAP in MDS patients obviously increases, the relationship between expression level of RAP1GAP and laboratory hematological parameter and IPSS score does not be confirmed. The role played by RAP1GAP expression in the pathogenesis of MDS and its clinical significance during progression of MDS towards AML deserves further studies. PMID- 19549375 TI - [Preliminary study on difference of Id4 gene methylation in various types of myelodysplastic syndromes]. AB - The aim of this study was to investigate the difference of Id4 gene promoter methylation in patients with different subtypes of myelodysplastic syndromes (MDS). By using MS-PCR method, the methylation status of Id4 gene was detected in 50 patients with different subtypes of MDS. Id4 methylation was also detected in bone marrow samples from patients with iron deficiency anemia which served as control. The results showed that Id4 gene was unmethylated in all of bone marrow samples from controls. In various subtypes of MDS patients, the rate of Id4 gene methylation was different. No Id4 methylation was found in 6 cases of RA, 2 cases of RARS and 4 cases of MDS-U. Id4 methylations was found in 2 out of 18 patients with RCMD. Id4 methylation in 3 out of 12 patients with RAEBI and other 3 out 8 patients with RAEBII were found. In groups with blast ratio lower or higher than 5%, the incidence of Id4 gene methylation were 6.7% and 30% respectively, so that there was significant difference. In tentative conclusion, Id4 gene methylation possibly is found in MDS patients with higher ratio of blast cells. PMID- 19549376 TI - [Changes of T-cell clonality after induction-cultivation of peripheral T lymphocytes in adoptive immunotherapy for leukemias]. AB - This study was purposed to analyze the changes of T-cell clonality after induction of peripheral T lymphocytes by autogenous DC and cytokines in the preparation of adoptive immunotherapy for leukemias. The bone marrow and peripheral blood from 21 leukemia patients at remission stage after treatment and subjected to adoptive immunotherapy were collected. Their DCs and T-cells were stimulated with cytokines and then were mixed to activate T-cells. T-cell receptor beta variable region (TCRBV) families were amplified by RT-PCR, and genescan method and sequencing of the PCR products were used to observe the clonality changes of T-cells before and after the induction and cultivation of T cells. The flow cytometry was used to identify CD3(+), CD4(+), CD8(+), CD3(+)CD56(+) and CD4(+)CD25str(+)FOXP3(+) cells to disclose the ratio change of cytotoxic T lymphocytes (CTL), helper T-cells, regulatory T-cells and NK T-cells before and after induction and cultivation of T-cells. The results showed that in the 21 patients, most of the 24 TCRBV families presented as oligoclonal distribution on genescan, several families were not expressed, and only a few families remained polyclonal. TCRBV24 was found to be oligoclonal in all of the 21 patients. DNA sequence analysis of TCRBV24 revealed a common motif of VAG in CDR3 in 3 cases and a common motif of GGG in CDR3 in 2 cases. In patient 5, both TCRBV 24 and TCRBV8 contained the same motif of GGG in CDR3. The identical motif in these patients may suggest that these T-cells recognize the same antigen. The peripheral lymphocytes demonstrated recovery of clonal profile on genescan from oligoclonal profile and absence of several families before the induction and cultivation to typical polyclonal profile in all TCRBV families after the induction by DC and cytokines for 13 days. After the induction and cultivation, the number of lymphocytes increased to 3.38 +/- 1.20 times. CD3(+), CD4(+), CD8(+), CD3(+)CD56(+) and CD4(+)CD25str(+)FOX P3(+) cells were 71.1 +/- 11.8%, 26.7 +/- 11.4%, 35.7 +/- 12.9%, 3.1 +/- 1.6% and 0.12 +/- 0.1% respectively before the induction and cultivation, and changed to 95.4 +/- 3.2% (p < 0.01), 27.0 +/- 13.1% (p > 0.01), 55.5 +/- 13.8% (p < 0.01), 9.8 +/- 6.1% (p < 0.01) and 0.22 +/- 0.18% (p < 0.01) respectively after the induction and cultivation. It is concluded that the major action of this induction and cultivation method on T lymphocytes in vitro is the promotion of CTL and NK T-cell proliferation. In leukemic patients at the remission stage, the TCRBV profile is characterized by the oligoclonal proliferation of T-lymphocytes. Several proliferated clones may have the same motif in CDR3, suggesting the recognition of the same antigen by these lymphocyte clones. Cytokine induction and co-culture with autogenous DCs can stimulate the T-lymphocytes to recover their immunocompetence as manifested by the polyclonal profile and the proliferation of CTL and NK-T cells. PMID- 19549377 TI - [Effects of interleukin 21 on anti-leukemia activity of cytotoxic T lymphocytes induced by dendritic cells]. AB - This study was aimed to explore the effects of interleukin 21 (IL-21) on the anti leukemia activity of cytotoxic T lymphocytes (CTL) induced by dendritic cells (DCs) in vitro. The peripheral mononuclear cells from leukemia patients in complete remission were cultured with the specific cytokines to induce the production of DCs. The DCs loaded with RNA from autologous leukemic cells as antigen, and co-cultured with autologous T lymphocytes to get leukemia specific CTL. The cytotoxic activity of CTL against autologous leukemic cells was measured by LDH release method. The concentration of IFN-gamma and TNF-alpha in the culture supernatant was measured by enzyme immunoassay. The effects of IL-21 on the mature DCs were also studied by the measurement of the phenotype of DC and the allogenic mixed lymphocytic reactions induced by DCs. Experiments were divided into 2 groups: test group in which IL-21 (200 ng/ml) was added in coculture of DC/CTL and control group in which no IL-21 (200 ng/ml) was added. The results showed that when cultured with IL-21, the quantity of CTL increased from (56.73 +/- 10.21)% (control group) to (73.43 +/- 18.01)% (p < 0.01); The concentration of IFN-gamma and TNF-alpha in the culture supernatant increased from (154.91 +/- 67.20) ng/L (control group) to (310.62 +/- 141.15) ng/L (p < 0.01) and from (8.77 +/- 5.09) microg/L (control group) to (15.25 +/- 6.56) microg/L (p < 0.01) respectively. At the effector: target ratio of 20:1, the cytotoxic activity against autologous leukemic cells by CTL increased from (50.22 +/- 5.07)% (control group) to (75.38 +/- 9.47)% (p < 0.01). IL-21 had neither effect on the phenotype (CD1a, CD83, CD86, CD80 and HLA-DR) of mature DCs nor the allogeneic mixed lymphocytic reactions induced by DCs. It is concluded that IL-21 can strengthen the proliferation of CTL, and improve the production of IFN-gamma and TNF-alpha, thus enhance the anti-leukemia activity of CTL. Nevertheless, there is no effect of IL-21 on the function of mature DCs. These data indicate that IL-21 has a potential clinical value in the enhancement of anti-leukemia immunotherapy. PMID- 19549378 TI - [Expansion and cytokine secretion profile of human valpha24(+) NKT cells from different sources]. AB - This study was purposed to investigate the phenotype, in vitro expansion and cytokine secretion profile of Valpha24(+) NKT cells from cord blood (CB), peripheral blood (PB), and granulocyte colony stimulating factor-mobilized peripheral blood mononuclear cells (G-PBMNCs). Fresh mononuclear cells (MNCs) were isolated by the method of gradient centrifugation and then cultured with alpha-GalCer (100 ng/ml), IL-2 (50 U/ml), IL-15 (50 ng/ml) for 12 days. Valpha24(+) NKT cells were purified by anti-Vbeta11 TCR McAb and goat anti-mouse IgG magnetic beads. The phenotype and purity of Valpha24(+) NKT cells were determined by flow cytometry. Cytokine production was analyzed by ELISA. The results showed that Valpha24(+) NKT cells in CB, PB and G-PBMNCs were expanded by 221.5 (95 - 501), 456.5 (101 - 2207), and 756.38 (82 - 20373)-fold respectively. After stimulation by phorbol-12-myristate-13-acetate (PMA) for 24 hours, IL-4 and IFN-gamma produced by Valpha24(+) NKT cells from CB and PB were 180.33 (144.67 - 2253.48) vs 190.67 (110.07 - 6060.16) ng/ml, 864.33 (401.33 - 3386.67) vs 508.49 (253.82 - 8840.00) ng/ml respectively, with IL-4/IFN-gamma ratio of 0.503 +/- 0.642 vs 0.455 +/- 0.562 respectively. After expansion of Valpha24(+) NKT cells from G-PBMNCs, IL-4 and IFN-gamma produced by Valpha24(+) NKT cells at day 9 and day 12 were 139.08 (7.62 - 606) vs 89.3 (0 - 729.2) ng/ml, 14264.8 (1168 - 18059) vs 14488 (1041 - 18261) ng/ml respectively, with IL-4/IFN-gamma ratio of 0.0531 +/- 0.1081 vs 0.0376 +/- 0.1148 respectively. It is concluded that in presence of IL-2 and IL-15, alpha-GalCer can facilitate the rapid short-term expansion of Valpha24(+) NKT cells from CB, PB, and G-PBMNCs. Valpha24(+) NKT cells from G PBMNCs show much high potential of expansion in comparison to the counterparts from CB or PB (p < 0.05). The activated Valpha24(+) NKT cells can secrete IFN gamma and IL-4 in large amounts, with IFN-gamma in particular. PMID- 19549379 TI - [Gene KIR in match with HLA-Cw impacts on NK cell cytotoxicity]. AB - The aim of this study was to investigate how the killer immune globulin-like inhibition receptor (KIR) in match with HLA-Cw impacts on NK cell activity. Mononuclear cells were isolated in 20 ml peripheral blood from 27 healthy persons by Ficoll-Hypaque and purified by NK cell isolation kit. Target cells were mononuclear cells isolated from bone marrow of 30 de novo AML patients. The KIR expression were detected by flow cytometry with antibodies against CD158a, CD158b. The 2 ml of peripheral blood from healthy persons and AML patients were collected, the DNA was extracted by using PROTRANS method, the HLA-Cw and KIR gene were detected by PCR-SSP typing with sequence specific primers. The NK cell cytotoxicity against AML cells was determined by MTT after combination of KIR with HLA-Cw gene. The results indicated that the purity of NK cells was (90.8 +/- 6.08)%. The less the KIR/HLA-Cw matched, the more activity was shown in NK cells. When no match of NK cell/target cell (KIR/HLA-Cw) there was, the cytotoxicity was (50.66 +/- 8.40)%, 1 or 2 matches showed cytotoxicity of (38.28 +/- 6.71)% and (19.74 +/- 4.15)% (p < 0.001). Expression level of KIRs on NK cells also was related with cytotoxicity level (p < 0.001). It is concluded that the interaction between inhibitory KIR and HLA ligands, and also expression level of KIRs on NK cells both impact significantly on NK cell function, that the less match of KIR/HLA-Cw, and the less expression of KIRs on NK cells result in the stronger NK cell cytotoxicity. PMID- 19549380 TI - [Construction and verification of a novel triple-regulated oncolytic adenovirus carrying gene Pdcd5]. AB - The purpose of this study was to construct a recombinant conditionally replicating adenovirus (CRAd) expressing programmed cell death 5 (pdcd5). Pdcd5 gene was inserted in the E3 region of SG600-a CRAd in which the key genes for virus replication E1a and E1b were controlled under the human telomerase reverse transcriptase promoter (hTERTp) and the hypoxia response element (HRE) respectively, and with a deletion of 24 nucleotides within CR2 region of E1a. The insertion and orientation of all recombined plasmids were confirmed by restriction enzyme digestion and polymerase chain reaction (PCR). The infection efficiencies of a recombined virus carrying enhanced green fluorescent protein (EGFP) in leukemic cell lines were observed by using fluorescence microscope. The relative pdcd5 expression levels of K562 after being infected with SG611-pdcd5 were detected by real-time quantitative PCR. The results showed that the construction of SG611-pdcd5 was completed and confirmed. Pdcd5, hTERTp, HRE, skeleton and fiber11 of recombinant adenovirus SG611-pdcd5 were successfully amplified. The infection efficiencies of SG611-EGFP were all above 70% in both leukemic K562 and MEG-01 cell lines. SG611-pdcd5 expressed pdcd5 with high efficiency in leukemic cells as compared with Ad-pdcd5 or SG611 (p < 0.001). The expression level of pdcd5 increased gradually along with the increase of MOI. It is concluded that the triple-regulated adenovirus of SG611-pdcd5 containing the pro-apopro-tic gene pdcd5 has been successfully established with high pdcd5 expression level in leukemic cells, indicating that the recombinant adenovirus, SG611-pdcd5, promises further development of targeted tumor gene therapy. PMID- 19549381 TI - Construction of ICAM-1-GFP and its binding with Molt-4 cells. AB - This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering. PMID- 19549382 TI - [Effect of demethylation treatment on the expression of inhibitory receptor KIR gene in NK-92MI cell line]. AB - The aim of this study was to analyze the promoter methylation patterns of inhibitory killer cell immunoglobulin-like receptor (KIR) which gene expression and the effect of demethylation treatment were studied, and to explore the possible regulation mechanism of inhibitory kir gene expression. The promoter methylation levels of kir2DL1 and kir2DL2/kir2DL3 in NK-92MI cell line were detected by bisulfite sequencing technique. Then NK-92MI cells were treated with 5-azacytidine to induce the demethylation of CpG islands. The levels of gene expression of kir were determined by RT-PCR. The results demonstrated that the methylation frequencies of CpG dinucleotides surrounding the promoter regions of kir2DL1 and kir2DL2/kir2DL3 genes were 25% to 88% and 5% to 80% respectively. DNA demethylating treatment with 5-azacytidine resulted in re-expression of kir2DL1 gene and increased expressions of kir2DL1, kir2DL2 and kir2DL3 genes in NK-92MI cells. In conclusion, the promoter DNA methylation participates in the regulation of kir gene expression in NK-92MI cells. PMID- 19549383 TI - [Role of phosphatidylinositol 3-kinase and myosin light chain kinase during the activation of thrombin receptors]. AB - The objective of study was to compare the influences of wortmannin on platelet aggregation and platelet membrane surface glycoproteins GPIb expression after thrombin receptor activation, and to investigate the role of phosphatidylinositol 3-kinase (PI3-K) and myosin light chain kinase (MLCK) in the course of thrombin receptor activation. Peptide SFLLRN (PAR1-AP) and AYPGKF (PAR4-AP) were used for stimulating platelet, and the changes of platelet aggregation and GPIb were analyzed with 100 nmol/L wortmannin (inhibitor of PI3-K) and 10 micromol/L wortmannin (inhibitor of MLCK). The results indicated that the platelet activation was influenced by either concentration of wortmannin in response to PAR stimulation. Platelet aggregation was apparently inhibited by 10 micromol/L wortmannin through both PAR peptides, and was slightly inhibited by 100 nmol/L wortmannin only under PAR1-AP activation. In addition, GPIbalpha internalization was partly inhibited by 100 nmol/L wortmannin in response to PAR1 (p < 0.05 at 1, 2, 5 min) and PAR4 (p < 0.05 at 2, 5, 10 min) activation. Meanwhile, 10 micromol/L wortmannin induced little change for GPIbalpha centralisation in the course of PAR activation, with a delayed restoration of surface GPIbalpha observed under PAR1-AP activation, and no change of GPIbalpha redistribution existed under PAR4-AP activation. It is concluded that the different roles of PI3 K and MLCK exist in the course of thrombin receptor activation. PI3-K accelerates the short course of GPIb centralisation for two PAR signal pathways, while MLCK inhibits the restoration of GPIbalpha in PAR1 pathway. PMID- 19549384 TI - [Quantitative analysis for JAK2 mutation in 98 patients with essential thrombocythemia and its clinical significance]. AB - The objective of this study was to identify the frequency and types of JAK2V617F mutation in chinese patients with essential thrombocythemia (ET), to quantitatively detect the level of mutation transcripts and to investigate its clinical significance. The frequency and types of JAK2V617F mutation were detected by amplification-refractory mutation sequencing polymerase chain reaction (ARMS-PCR), the transcript level of JAK2V617F mutation was determined by using capillary electrophoresis. The results indicated that the JAK2V617F mutation was detected in 59 out of 98 patient with ET, 18 of whom were homozygous mutation. The mean age of patients with homozygous and heterozygous mutation was higher than that of patients with wild type mutation (p < 0.05). The quantitative assay using capillary electrophoresis showed that the transcript level of JAK2V617F mutation in patients with homozygous mutation was (89.9 +/- 6.7)%, which was higher than that in patients with heterozygous mutation (57.1 +/- 6.7)% (p < 0.05); the transcript level of JAK2V617F mutation in patients with age < 60 years was (62.3 +/- 16.5)%, which was lower than that in patients with age > 60 years (72.4% +/- 15.8)% (p < 0.05). The rate of thrombotic complications in patients with JAK2V617F-positive was higher than that in patients with JAK2V617F negative in which the rate of thrombotic complication in patients with homozygous mutation was higher than that in patients with heterozygous mutation (p < 0.05). Compared with patients without thrombotic events, there were higher level of transcripts of JAK2V617F mutation in patients with thrombotic events. It is concluded that the JAK2V617F positive and negative patients with ET display the different clinical features, therefore, the analysis of mutation types and detection of transcript levels not only helps to identify the disease status and progression, but also guides the treatment of ET patients. PMID- 19549385 TI - [Influence of long-used staphylokinase derivative on hemoagglutinative and fibrinolytic systems]. AB - This study was aimed to explore the influence of staphylokinase derivative (SAKD) on the hemoagglutinative and fibrinolytic systems, and to determine the safety of the staphylokinase derivative in application. The normal and model rats each 30 were divided into normal saline, SAKD and rSAK groups. The hemorrhage, bleeding time (BT), blood platelet count (BPC), activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen (Fg), D-dimer (D D), plasminogen (PLG) and plasmin inhibitor activity (PI) were detected before and after the administration with staphylokinase derivative 0.5 mg/kg body weight, once three days for consecutive 15 days. The results indicated that one case of normal rats with SAKD and two cases of high fat diet model group had mild hemorrhage, all of which showed automatic hemostasis; and 3 cases in rSAK group had mild hemorrhage. And the platelet counting, D-D, PLG and PI in all groups did not significantly change. The rats of high fat diet group treated with SAKD showed the significant extension of APTT, PT and TT times, and the decrease of Fg time (p < 0.05). All the experimental results demonstrated that the influence of SAKD on the hemagglutination of the normal animals was lower, however, which can improve the high-hemagglutination status of the rats with high fat diet. It is concluded that the SAKD at the dosage of this study has the higher safety, which can alleviate the high hemagglutination symptoms of the rats with high fat diet. PMID- 19549386 TI - [Analysis on promoter CpG methylation and expression of HOXB4 gene in cord blood CD34(+) cells and peripheral blood mononuclear cells]. AB - This study was aimed to examine the expression and promoter CpG island methylation of homeobox B4 (HOXB4) gene in CD34(+) cells from cord blood and peripheral blood mononuclear cells (PBMNCs) from health adult, and to investigate the expression level of HOXB4 in these two cells and its relationship with the promoter methylation. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to examine the expression of HOXB4 in CD34(+) cells and PBMNCs, and bisulfite sequencing technique was used to detect the methylation status of the promoter CpG sites of HOXB4 gene in CD34(+) cells and PBMNCs. The results indicated that highly expressed HOXB4 and unmethylation of HOXB4 promoter CpG island occurred in CD34(+) cells. However, loss of HOXB4 expression and the methylated CpG island of HOXB4 were observed in PBMNCs, and the methylated C residue was positioned at -129 bp in the upstream of ATG. It is concluded that the methylation status of HOXB4 gene promoter may be one negative regulatory mechanism for HOXB4 gene expression. The unmethylation of CpG island in the promoter region of HOXB4 gene may be correlated with the high expression of HOXB4 gene in CD34(+) cells, while the promoter methylation of HOXB4 gene may be associated with HOXB4 gene silencing in PBMNCs. The preliminary identification of HOXB4 promoter methylation site would provide a basis for further study and a novel approach to expand hematopoietic progenitor cells. PMID- 19549387 TI - [Transfection of bone marrow mesenchymal stem cells by adeno-associated virus 2/1 vector]. AB - This study was aimed to investigate the transfection efficacy of recombinant adeno-associated virus 2/1 (rAAV2/1) on bone marrow mesenchymal stem cells (BMMSCs) at different multiplicities of infection (MOI) and time, and effect of transfection on growth of rat BMMSCs. The rat BMMSCs cultured in vitro were transfected by using rAAV2/1 with enhanced green fluorescent protein (rAAV2/1 EGFP) at MOI of 1 x 10(4), 1 x 10(5) and 1 x 10(6); the EGFP expression was observed by fluorescent microscopy at 3, 7 and 14 days. The viability, proliferation multiple, differentiation ability of daughter cells were detected for evaluating the effect of rAAV2/1 on survival, proliferation and differentiation of BMMSCs and the fluorescence index (FI) were determined by flow cytometry. The results indicated that after transfection with rAAV2/1 for 24 hours the green fluorescence in BMMSCs were observed, but also the fluorescence gradually was enhanced along with prolonging of time, and reached to steady level after 7 days; the viability, proliferation multiple, differentiation ability of BMMSCs transfected by rAAV2/1-EGFP at different MOI showed no significant changes at 3,7 and 14 days (p > 0.05), meanwhile at same MOI the proliferation multiple obviously increased in comparison between 7 day vs 3 day and 14 days vs 7 days (p < 0.01). The flow cytometric detection showed that the transfection efficacy of rAAV2/1-EGFP on BMMSCs and FI increased significantly as the multiplicity of infection and culture time increased (p < 0.05). It is concluded that rAAV2/1 EGFP is able to transfect into BMMSCs effectively, but the transfection efficiency and fluorescence index increase significantly along with increase of multiplicity of infection and culture time. rAAV2/1-EGFP do not affect viability, proliferation multiple and differentiation ability of BMMSCs. rAAV2/1 is a kind of active vector for gene transfer to reform BMMSCs. PMID- 19549388 TI - [Exogenous gene expression in vitro and in vivo in bone marrow mesenchymal stem cells modified by hPDGF-A and hBD(2)]. AB - The objective of this study was to investigate the expression of exogenous hPDGF A and hBD(2) in gene-modified bone marrow mesenchymal stem cells (BM-MSCs) in vitro and in vivo. Recombinant adenovirus vector expressing hPDGF-A/hBD(2) genes was constructed and packaged into virion. Primary isolated and cultured BM-MSCs were transfected by using hPDGF-A hBD(2), then the expressions of exogenous hPDGF A/hBD(2) were detected by immunocytochemical staining in vitro. The conditioned medium (serum-free cultured supernatant of BM-MSCs transfected with recombinant adenovirus) collected from gene-modified BM-MSCs was applied to scratch wound on monolayer cells of multipotential cell line 10T1/2 in order to confirm the stimulative effect of hPDGF-A on cell migration. Gene-modified BM-MSCs were topically transplanted on wound of rats with radiation and skin excision combined injury. The distribution of BM-MSCs and expression of hPDGF-A/hBD(2) on the wound was observed by fluorescent microscopy and immunohistochemical staining respectively. The results indicated that the rat BM-MSCs transfected with recombinant adenovirus could express the EGFP in vitro. The immunofluorescent cytochemistry assay showed that the gene-modified BM-MSCs expressed the hPDGF-A and hBD(2). The scratch test confirmed that the percentage of healing area of wound in cultured supernatant group of gene-modified BM-MSCs was significant higher than that in control group on 8, 12, 24 and 48 hours (p < 0.05). The fluorescence microscopy of exogenous gene-modified BM-MSCs transplanted on wound revealed that the gene-modified BM-MSCs could higher express exogenous genes of EGFP at least within 2 weeks. The immunohistochemistry staining of wound indicated that the expression of exogenous genes began from day 3, reached to peak on day 7, and still visible on day 21 even though the expression became weak because of the possible dilution of the exogenous genes during cell division. It is concluded that efficient expression of exogenous hPDGF-A/hBD(2) in gene modified BM-MSCs are demonstrated both in vitro and in vivo, which suggests that the molecular mechanism underlying chronic wound-healing accelerated by the strategy combining cell therapy with gene therapy. PMID- 19549389 TI - [Transfection of gene mdr1 into human bone marrow mesenchymal stem cells by lentiviral vector]. AB - This study was aimed to investigate the feasibility and security of mdr1 gene modified mesenchymal stem cells (MSCs) so as to establish the experimental foundation for gene therapy. Lentiviral system was utilized to introduce the mdr1 gene into MSCs which were isolated from human bone marrow and cultured in vitro; RT-PCR and GFP marker were used to determine the expression of mdr1; MTT and trypan blue staining were used to detect the proliferative capacity of the MSCs. The results indicated that MSCs were infected with lentivirus at a multiplicity of infection (MOI) of 10 with optimal expression efficiency of 80%; the expressions of CD34, HLA-DR, CD31 and CD45 on surface of MSCs were found at low levels, however, the expressions of CD44, CD105, CD90 and CD13 on surface of MSCs were observed at high levels; GFP marker was observed on 72 hours after gene transfection and then gradually was enhanced; the expression of mdr1 mRNA appeared in transfected cells; Mdr1 transfection did not show a significantly inhibitory effect on MSCs. It is concluded that the expression of mdr1 is up regulated in MSCs transfected successfully by lentiviral vector, and the transfection has no significantly effects on survival and proliferation of MSCs. PMID- 19549390 TI - [Expression of Toll-like receptors in human bone marrow mesenchymal stem cells]. AB - The aim of this study was to explore the characteristics of Toll-like receptor expression in mesenchymal stem cells derived from bone marrow of healthy donor (BM-MSCs). BM-MSCs were isolated from bone marrow of healthy donor by Ficoll method. Expressions of CD34, CD45, HLA-DR, CD44 and CD71 in BM-MSCs were detected by flow cytometry. CD71 in BM-MSCs was assayed by immunocytochemistry. The adipocyte and osteoblast induction of BM-MSCs were detected by alizarin red stain and oil red stain respectively. TLR 1 - 10 mRNA levels in BM-MSCs were evaluated by semiquantitative RT-PCR. The results showed that expressions of CD34, CD45 and HLA-DR in BM-MSC were negative while the expressions of CD44 and CD71 were positive. CD71 in BM-MSCs was positive. After induced by osteoblast and adipocyte inductor, BM-MSCs were positive for alizarin red staining and oil red staining respectively. All of TLR 1 - 10 mRNA were found in BM-MSCs with high expression levels of TLR2, TLR3, TLR4, TLR7, TLR8, TLR9 and low expression levels of TLR1, TLR5, TLR6, TLR10. In conclusion, different levels of TLR 1 - 10 mRNA were expressed in BM-MSCs of healthy donor. PMID- 19549391 TI - [Transfusion of allogeneic mesenchymal stem cells promotes progression of atherosclerotic plaque in rabbits]. AB - The present study was purposed to evaluate the safety of mesenchymal stem cell (MSC)-based therapy impacting on atherosclerosis. Allogeneic MSCs were obtained from rabbit bone marrow aspirates and expanded in vitro. New Zealand white rabbits were divided into three groups: 24 rabbits with hypercholesterolemia receiving intravenous injection of either 5 x 10(7) MSCs (n = 12) or saline (n = 12) after 5 weeks on a high lipid diet and additional rabbits (n = 6) fed with standard rabbit diet were served as controls. Body weight and blood lipids were measured at weeks 0, 5, 9 and 13 during the study. All rabbits were sacrificed at week 13. Atherosclerotic lesion size and vasa vasorum were evaluated by using pathological analysis and immunocytochemical technique. The results showed that the aortic sinus lesion size significantly increased in rabbits infused with MSCs as compared with controls receiving saline (23.35 +/- 3.51% and 11.39 +/- 3.08% respectively). The lesion size in whole aortas of MSC-treated rabbits was 76.64 +/- 12.70% versus 57.61 +/- 9.00% in saline-treated animals (p < 0.05). Moreover, vasa vasorum networks in MSC-treated aortas were more numerous and had increased capillary density. It is concluded that the allogeneic MSC transfusion may result in an increase in atherosclerotic lesion size. In cell therapy with MSCs or cell populations containing MSCs a strategy to attenuate the high potential of MSCs involved in atherogenesis of atherosclerosis should be taken in account. PMID- 19549393 TI - [Analysis of risk factors for overall survival at 5 years in 96 patients after allogeneic hematopoietic cell transplantation]. AB - The aim of this study was to analyze the risk factors for overall survival at 5 years in 96 patients undergoing allogeneic hematopoietic stem cell transplantation by retrospective analysis. 11 clinical parameters including age, sex, disease status, HLA locus, donor type, donor-recipient blood type, conditioning regimen, aGVHD, HC, VOD and IP were selected for univariate analysis by using a Cox regression. Factors have statistic significance at the 0.1 level on univariate analysis were evaluated by multivariate analysis by a Coxs regression. The cumulative incidence of aGVHD and survival rate of patients were calculated by the method of Kaplan and Meier. The results showed that 95 patients achieved sustained donor engraftment except 1 patients. The median time of leukocyte engraftment (ANC > or = 0.5 x 10(9)/L) was 13 days. The aGVHD of I - IV grade was observed in 42 out of 96 patients (43.75%), in which 11 patients were with aGVHD of I grade (11.46%), 19 patients were with aGVHD of II grade (19.79%), 12 patients were with aGVHD of III - IV grade (12.50%). Out of 96 patients 10 relapsed and 38 dead, the overall survival at 5 years was 60.42%. The Cox regression analysis showed that aGVHD and disease status before transplant were main factors affecting long-term survival of patients, relative risks of which were 2.996 and 2.619 respectively. It is concluded that the main factors affecting long-term survival of patients are aGVHD and disease status. The key to improve the outcome of allo-HSCT is to reduce the incidence and severity of aGVHD, meanwhile to select the CR1 for allo-HSCT to treat the patients in advanced refractory and relapsed situation should be considered as important risk factors. PMID- 19549392 TI - [Short tandem repeat panel settled for quantitative chimerism analysis following allogeneic stem cell transplantation and its application]. AB - To evaluate the roles of 8 short tandem repeats (STR) loci as STR panel in quantitative analysis of chimerism following transplantation, the primers were synthesized and marked with different dyes for D3S3045, D4S2366, D4S2639, D5S818, D13S317, D18S1002, D20S481 and D22S689. The blood samples of 15 cases received allogeneic stem cell transplantation were collected before and after transplantation, then DNA was extracted and amplified with these primers, and was further analysed under ABI Genetic Analyser 3100 to select suitable informative STR locus. Donor/recipient dilution series were prepared to get standard curves in selected loci, the DNAs extracted at different days after transplantation were used to quantitatively analyze the chimerism in patients according to the values of peak area or peak height of fluorescent signals. The standard curves can be used to calculate the chimerism by plotting the respective R/D quotient value against the percentage of recipient DNA. The results indicated that the calculated chimerism was in concordance with the donor/recipient dilution. The STR panel succeeded in identifying at least one informative marker and quantitative monitoring the chimerism after HSCT in 15 donor-recipient pairs and a relapsed case was diagnosed. It is concluded that the STR panel and its detection method can accurately and quantitatively monitor the chimerism after allogeneic HSCT, which is more economical and flexible than using commercial kits. PMID- 19549394 TI - [Allogeneic hematopoietic stem cell transplantation in 6 patients with myelodysplastic syndrome]. AB - This study was purposed to explore the efficacy of hematopoietic reconstitution and survival of patients with myelodysplastic syndrome (MDS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Allo-HSCT without T lymphocyte depletion was used in 6 patients with MDS from November 1999 to June 2007. 4 cases out of them received allo-PBSCT from HLA matched sibling donors with conditioning regimen of cyclophosphamide (CTX) and Bu. Graft versus host disease (GVHD) was prevented by the administration of immunosuppressive drugs of cyclosporine A (CsA) and short-course MTX. 2 patients received haploidentical allogeneic bone marrow transplantation (hi-alloBMT) after preconditioning with cytosine arabinoside (Ara-C), CTX and total body irradiation (TBI) with a linear accelerator. GVHD was prevented by the administration of immunosuppressive drugs including CSA, short-course MTX, MMF, anti-CD25 monoclonal antibody and ATG. The results showed that all of the patients were engrafted successfully. The median time of granulocyte recovery exceeding 0.5 x 10(9)/L and platelets exceeding 20 x 10(9)/L were days 15 and 20.3 respectively, and 100% donor hematological cells were detected by cytogenetic analysis. All patients did not experience serious acute graft-versus-host disease (aGVHD). During 18 - 108 months of following-up, 2 cases died of pulmonary complication and of relapse; the other 4 cases survive in a disease-free situation. In conclusion, allo-HSCT was an effective approach for the treatment of MDS. PMID- 19549395 TI - [Clinical analysis of acute renal failure after allogeneic hematopoietic stem cell transplantation]. AB - The aim of this study was to investigate the incidence, risk factors of acute renal failure (ARF) after allogeneic hematopoietic stem cell transplantation (allo-HSCT), and evaluate its effect on the prognosis of patients after allo HSCT. A retrospective analysis was performed in 86 patients undergoing allo-HSCT at Peking University First Hospital from June 2003 to April 2007. ARF is defined as a doubling of baseline serum creatinine at any time during the first 100 days post-transplant. The risks of ARF and mortality after ARF were examined using univariate analysis and multivariate unconditional logistic regression. The correlation of ARF and survival was examined using Cox regression. The results indicated that 27 patients (31.40%) developed ARF at a median of 59.5 days after transplant (range 1 to 93 days). The univariate analysis showed that elevated risks were severe acute GVHD (OR 6.196; 95% CI 1.121 - 34.249, p = 0.033), sepsis or septic shock (OR 4.184; 95% CI 1.314 - 13.325, p = 0.018) and hyperbilirubinemia (OR 3.709; 95% CI 1.428 - 9.635, p = 0.006). Renal disease before transplant (OR 6.711; 95% CI 1.199 - 37.564, p = 0.027), hypertension (OR 2.067; 95% CI 0.739 - 5.782, p = 0.165), the use of vancomycin (OR 2.133; 95% CI 0.844 - 5.392, p = 0.106) or foscarnet sodium (OR 2.133; 95% CI 0.844 - 5.392, p = 0.106) may be potential risks. Multivariate logistic regression analysis showed that renal disease before transplant (OR 6.288; 95% CI 1.218 - 32.455, p = 0.028), sepsis or septic shock (OR 3.614; 95% CI 1.040 - 12.544, p = 0.043) and hyperbilirubinemia (OR 4.448; 95% CI 1.563 - 12.665, p = 0.005) appear to be independently associated with an increased risk of ARF. Age, gender, baseline serum creatinine level, advanced malignant disease, unrelated-donor, total body irradiation (TBI) and cyclosporine levels were not associated with the development of ARF. Cox regression showed that ARF (RR 2.124; 95% CI 1.016 - 4.441, p = 0.045) was independently associated with survival of patients after allo-HSCT. The mortality of patients with ARF within 6 months post-transplant was significantly higher than that of those without ARF (44.4% vs 8.47%, p < 0.001). It is concluded that the cumulative incidence of ARF after allo-HSCT remains high. Renal disease before transplant, hyperbilirubinemia and sepsis or septic shock are all related factors which can increase the risk of ARF. ARF appears to be independent factor influencing survival of patients after allo-HSCT. PMID- 19549396 TI - [Relationship between polymorphisms of FCGR2B and susceptibility of children idiopathic thrombocytopenic purpura]. AB - The aim of study was to investigate the relationship between polymorphisms of FCGR2B232 1/T oligonucleotide and the susceptibility of children with idiopathic thrombocytopenic purpura (ITP). DNA from 76 patients with ITP and 37 controls was extracted. The SNPs of FCGR2B-232 was detected by polymerase chain reaction (PCR) combined with direct sequencing. The genotype distribution and allele frequency among different groups were compared. The results showed that the genotype (I/I, I/T, T/T) of FCGR2B-232 were 55.3%, 42.1%, and 2.6% in 76 patients with ITP, while 81.1%, 18.9%, 0% in 37 controls. The allele frequencies of FCGR2B-232 in patients with ITP were 76.3% (I232) and 23.7% (T232), but 90.5% and 9.5% in controls. There were significant differences in genotype distributions between the ITP patients and controls (chi(2) = 7.45, = 0.024). The enrichment in Thr232 allele carrier was also significant among the ITP patients as compared with the controls (chi(2) = 7.18, p = 0.007, odds ratio 3.47). There were also significant differences in allele frequencies between the ITP patients and controls [chi(2) = 6.54, p = 0.011, odds ratio 2.97, 95% CI (1.25 - 7.05)]. It is concluded that the polymorphisms of FCGR2B-232 significantly correlates with the susceptibility of children suffering from ITP. The minor Thr232 allele may be a risk genetic factor to ITP children. PMID- 19549397 TI - [Biological characteristics of bone marrow-derived mesenchymal stem cells in children with acute leukemia]. AB - This study was aimed to investigate the conditions of culturing in vitro mesenchymal stem cells (MSCs) derived from bone marrow of children with acute leukemia and the biological characteristics of MSCs from leukemia children. The bone marrow MSCs of acute leukemia children were isolated by density gradient centrifugation combined with adherent segregating method and cultured in DMEM/F12. The morphology of Wright stained MSCs was observed under inverted microscope. Cell surface markers were analyzed with flow cytometry. The growth characteristic features of cultured MSCs was measured with MTT method. Induced adipogenic and osteogenic differentiation of MSCs in appropriate induction media was observed. The results indicated that BM-MSCs of acute leukemia children could be successfully cultured in vitro in appropriate conditions. At 24 hours of culture the MSCs began to adhere to wall, grew in colony and appeared in different shapes. As the culture lasted, the MSCs proliferated continuously and shaped in fusiform. After 2 - 3 weeks of culture, MSCs covered the bottom of culture flask. The analysis of growth feature showed that MSCs were in latency for 3 days, and then entered into growth period. After 8 days of culture the growth of MSCs showed to be in plateau stage. The shape of MSCs in 1st and 2nd generation showed to be heterogeneous but the 3rd generation to be homogeneous with long-fusiform. Cells were arranged in shape of whirlpool or radiation. The surface marker analysis showed that the MSCs were positive for CD105, CD29, CD13, but negative for CD34, CD45, CD14 and HLA-DR. The MSCs from leukemia children could be induced into adipocytes and osteocytes in appropriate conditions. It is concluded that (1) MSCs derived from children with acute leukemia can be successfully cultured and passaged in vitro; (2) MSCs from leukemia children not received chemotherapy are more successfully cultured in vitro than those received chemotherapy; (3) the common biological characteristics of MSCs from children with acute leukemia are same as the MSCs from healthy person. PMID- 19549398 TI - [Side effects of L-asparaginase during therapies for remission induction and maintenance in children with acute lymphocytic leukemia]. AB - This study was purposed to investigate the possible side effects of L asparaginase (L-ASP) in the treatment of patients with acute lymphoblastic leukemia (ALL) and to explore the correlation of these side effects at different therapeutic stages by means of retrospective analysis, so as to reduce the incidence of side effects and improve the safety of chemotherapy and the long term survival of patients. The probability and severity of side effects related to use of L-ASP in 38 cases of ALL during remission induction therapy (VDLDex regimen) and 40 cases of ALL during maintenance intensive therapy (VMLDex regimen) were compared. The results showed that allergic response, diabetes and drug-induced liver disease happened more frequently during maintenance therapy than during remission induction therapy, while defibrination, abnormal hemagglutinin, acute pancreatitis, hypoproteinemia, gastrointestinal reaction and infectious shock were observed more during remission induction therapy than those at maintenance therapy. In conclusion, the L-ASP showed some side effects especially for the patients during the remission induction therapy which should be paid enough attention. The regular and comprehensive monitoring can effectively reduce and avoid the side effects of L-ASP, to improve the safety of chemotherapy. PMID- 19549399 TI - [Correlation of JAK2V617F mutation burden with clinical features in patients with polycythemia vera and essential thrombocythemia]. AB - This study was aimed to analyze the correlation of JAK2V617F mutation burden with clinical features in patients with polycythemia vera (PV) and essential thrombocythemia (ET), The JAK2V617F mutation ratios in 47 PV samples and 43 ET samples were detected by real-time PCR. The correlation of mutation allele ratio in PV and ET samples with clinical features (hemoglobin, hematocrit, white blood cell count and platelet count) was analyzed. The results showed that the JAK2V617F mutation burden was higher in PV (0.441 +/- 0.270) than that in ET (0.209 +/- 0.192). The JAK2V617F mutation burden was positively correlated with levels of hemoglobin (PV: R = 0.518, p < 0.001; ET: R = 0.528, p = 0.005), hematocrit (PV: R = 0.510, p < 0.001; ET: R = 0.524, p = 0.005) and leukocyte (PV: R = 0.584, p = < 0.001; ET: R = 0.471, p = 0.013) in PV and ET samples. The higher JAK2V617F mutation burden was negatively correlated with levels of platelet count in PV samples (R = -0.354, p = 0.020), but there was no correlation between the JAK2V617F mutation burden and platelet count in ET samples (R = 0.233, p = 0.242). It is concluded that the higher JAK2V617F mutation burden is related with higher hemoglobin, hematocrit and leukocyte count in both PV and ET samples. The higher JAK2V617F mutation burden is correlated with lower platelet count in PV samples, but there is no correlation between JAK2V617F mutation burden and platelet count in ET samples. PMID- 19549400 TI - [Analysis of complicated anemia in 60 patients with systemic lupus erythematosus]. AB - The study was aimed to explore the characteristics of clinical manifestation, laboratory indicators and bone marrow examination of SLE patients with anemia. 60 SLE patients with anemia were analyzed for their clinical manifestation, laboratory indicators and bone marrow examination in comparison with 40 contemporaneous SLE patients without anemia. The results indicated that there were significant differences in clinical manifestations of fatigue between the SLE patients with anemia and those without anemia. The detection rate of the decreased Plt and C4 and the percentages of eosinophils, early normoblast, polychromatic normoblast and orthochromatic normoblast in bone marrow were all higher than that in those without anemia. The ANA with titer 1:320 in SLE patients complicated by anemia was lower than that in those without anemia. In conclusion, the clinical manifestation, experimental examination and bone marrow findings were significantly different between the SLE patients with anemia and without anemia. PMID- 19549401 TI - [Long-term survival analysis in 89 adult patients with acute myeloid leukemia of fusion gene aml1/eto positive]. AB - This study was aimed to investigate various factors influencing long-term survival in adult AML patients with fusion gene aml1/eto positive. A single institutional retrospective study with long-term follow-up was performed to better define the prognostic factors for AML patients with aml1/eto positive. Newly diagnosed 89 adult AML patients with aml1/eto positive were followed up for 1 to 42 months (median 24 months) from January 2004 to July 2008. Univariate and multivariate analysis of potential factors influencing survival and prognosis were carried out by using Log-Rank and Cox regression method, including sex, age, initial WBC counts, extramedullary leukemic disease, central nervous system leukemia (CNSL), chromosome aberrations, immunophenotype, first induction regimen, chemotherapy course to complete remission (CR), time from induction therapy to CR, negative or positive rate of aml1/eto and allogeneic hematopoietic stem cell transplantation and so on. The results showed that the estimated 5-year overall survival (OS) and relapse-free survival (RFS) were (50.0 +/- 2.3)% and (47.0 +/- 1.9)% respectively in follow-up of 89 patients for 1 - 42 months (mean 24 months). Univariate analysis revealed that initial WBC counts, CNSL, chemotherapy course to CR, time from induction therapy to CR, persistent negative in remission and allogeneic hematopoietic stem cell transplantation were important prognostic factors for long-term surviva1. Multivariate study demonstrated that initial WBC counts, CNSL, CD56 positive, negative or positive rate of aml1/eto, time from induction therapy to CR, persistent negative result of RT-PCR assay in remission and allogeneic hematopoietic stem cell transplantation were all critical factors in relation to OS and RFS. It is concluded that Chinese adult AML patients with fusion gene aml1/eto positive have some different characteristics as compared with patients from other countries, a relatively poor outcome is observed in patients, HSCT should be recommended to adult AML patients. PMID- 19549402 TI - [Secondary acute myeloid leukemia complicated after treatment of non-Hodgkin's lymphoma]. AB - The aim of this study was to investigate the mechanism, susceptibility, (18)F-FDG positron emission computerized tomography ((18)F-FDG PET/CT) features and the treatment of therapy-related acute myeloid leukemia. One patient with NHL was affected with t-MDS after treatment and then progressed to t-AML. The clinical data including bone marrow cell morphology, flow cytometry, karyotype and PET/CT features were analyzed. The results showed that the primary treatment for NHL refers to varieties of cytotoxic drug such as cyclophosphamide-hydroxydaunomycin oncovin-prednisone (CHOP) chemotherapy. The interval time from the chemotherapy of NHL to the occurrence of t-MDS was 105 months and t-MDS progressed to AML-M(2) in 2 months. Karyotype analysis results of t-MDS and t-AML were normal. (18)F-FDG PET indicated that the FDG uptake in the bone raised diffusely. The patient showed complete response after second-line therapy (CAG regiments). In conclusion, the occurrence of t-AML/MDS may be associated with the application of the cytotoxic chemotherapeutics. (18)F-FDG PET may be an indicator predicting the transformation of t-MDS to t-AML. PMID- 19549403 TI - [Low-dose cytarabine and aclarubicin in combination with granulocyte colony stimulating factor priming in 50 patients with relapsed acute myeloid leukemia]. AB - To evaluate the efficacy and toxicity of low-dose cytarabine and aclarubicin in combination with granulocyte colony-stimulating factor (G-CSF) protocol for patients with relapsed acute myeloid leukemia (AML). A total of fifty relapsed patients have been enrolled, including 13 early relapsed and 37 late relapsed. 24 patients were male and 26 were female, with age ranging from 15 to 69 (median 47) years. Out of them, 7 patients relapsed after allogeneic peripheral blood stem cell transplantation (allo-PBSCT), 3 patients relapsed after autologous peripheral blood stem cell transplantation (auto-PBSCT), 25 patients relapsed after received regimens including high dose cytarabine and 15 patients relapsed after CR or stopping chemical therapy themself in course of consolidatory therapy. 30 relapsed patients received CAG regimen, and 20 patients (control group) received an anthracycline in combination with cytarabine. The results indicated that after one course, the complete remission (CR) rate was 46.7% (14/30), the CR rate after allo-PBSCT was 50% (3/6), the early death rate was 3.3% in CAG group; and CR rate was 30% (6/20) and the early death rate was 15% in control group. Myelosuppression was mild to moderate, and no severe nonhematologic toxicity was observed in two groups. The overall median times in CAG group and control group were 22 and 19 months respectively. In conclusion, CAG regimen as the induction therapy is effective and well tolerable with low side effects for relapsed patients who had received high dose cytarabine, auto PBSCT or allo-PBSCT. PMID- 19549404 TI - [A preliminary investigation on early diagnosis of invasive aspergillosis in patients with blood diseases by using circulating galactomannan test]. AB - The objective of this study was to explore the useful value of circulating galactomannan (GM) for early diagnosis of invasive aspergillosis. All 141 patients were classified as 103 patients of clinical and possible diagnosis, and 38 non-Aspergillus patients. 209 serum samples for the detection of GM by Platelia Aspergillus were collected before anti-fungal vaccine therapy. ELISA method was used in detection of GM. The results showed that (1) the sensitivity of 87.5%, specificity of 81.6%, positive prediction of 66.7% and negative prediction of 93.9% were determined by using cut-off value. According to the result of ELISA, the clinical diagnosed patients was up to 48, while the possible diagnosed patients were 55. (2) Among 62 patients with consecutive examinations of serum samples, 50 patients were successfully diagnosed and treated, while 12 patients died. A progressive reduction of GM level was found in survivors, however, the patients of poor prognosis showed higher antigen titres. It is concluded that GM test has more significance for earlier diagnosis of aspergillosis, the concentration of GM is related to prognosis of disease. PMID- 19549405 TI - [Auto-hematopoietic stem cell transplantation for multiple myeloma accompanied with amyloidosis in four limbs]. AB - The study was aimed to investigate the effective therapeutic method for patients with multiple myeloma accompanied with amyloidosis. A 58-year-old patient diagnosed as multiple myeloma accompanied with amyloidosis in four limbs was enrolled in this study. The various clinical and laboratorial examinations were performed, including bone marrow smear, immunologic test, radiography and so on. Patient received chemotherapeutic drugs and then autologous hematopoietic stem cell transplantation (auto-HSCT). The result showed that hematopoietic reconstitution was achieved at 23 days after auto-HSCT. Immunofixation electrophoresis was normal. There was only 0.6% plasma cells in the bone marrow. In conclusion, the auto-HSCT may be an effective treatment for multiple myeloma accompanied with amyloidosis in four limbs. PMID- 19549406 TI - [Clinical study on fludarabine combined with cytarabine regimen in the treatment of patients with refractory and relapsed acute myeloid leukemia]. AB - The aim of study was to evaluate the clinical efficacy and toxicity of fludarabine combined with cytarabine (FA) regimen in the treatment of patients with refractory and/or relapsed acute myeloid leukemia (AML). Nineteen cases with refractory/relapsed AML were treated with FA regimen in which fludarabine phosphate 25 mg/(m(2) x d), d1-5; cytarabine (Ara-C) 2 g/(m(2) x d), d1-5. Another 20 cases were treated with salvage chemotherapy (MAE regimen: mitoxantrone, Ara-C and etoposide or DAE regimen: daunorubicin, Ara-C and etoposide). All patients received at least 2 cycles chemotherapy. The results showed that 9 patients (47%) in FA regimen group achieved complete remission (CR), 8 cases (42%) obtained partial remission (PR), the clinical efficacy was superior to that of the MAE or DAE regimens (p < 0.05). Major toxicity of FA regimen was myelosuppression. Grade IV hematologic toxicity occurred in all patients received FA regimen. Nonhematologic complications consisted of gastrointestinal side effects, mucositis, liver toxicity, which were mild to moderate and could be alleviated with supportive therapy. In conclusion, FA regimen is an effective regimen for treatment of refractory and relapsed AML. PMID- 19549407 TI - [Distribution of tap1 and tap2 loci allelic and genotypic frequencies in Chinese Hans]. AB - This study was aimed to investigate the distribution and implication of tap1 (transporter associated with antigen processing) and tap2 loci allelic and genotypic frequencies. The distribution of tap1 and tap2 loci allelic and genotypic frequencies in 339 random samples of healthy Chinese Hans was analyzed by TaqMan PCR. Several genetic information about power of discrimination, cumulative DP, polymorphism information content, expected heterozygosity and observed heterozygosity were calculated. The results indicated that 5 tap1 alleles (tap1*0101, 020101, 020102, 0301 and 0401) and 4 tap2 alleles (tap2*0101, 0102, 0103 and 0201) were detected in all samples. 8 tap1 genotypes were found which account for 53.3% of the theoretic genotype and 6 tap2 genotypes were found which account for 60% of the theoretic genotype. The genotyping results of tap1 and tap2 both conform to the Hardy-Weinberg expectations (p > 0.05). Tap1*0101 (79.79%) and tap2*0101 (82.74%) are the most common alleles in Chinese Hans. It is concluded that tap1*0101 and tap2*0101 are most common alleles in Chinese Hans, tap1 and tap2 loci carry some power of individual discrimination and polymorphism information content. These two locl can be used for the research in the fields of human genetics, linkage analysis of genetic disease genes, paternity test and individual identification and so on. PMID- 19549408 TI - [A total of 362 HLA different haplotypes and HLA recombination haplotypes based on analysis of their family pedigree in Chinese partial Han populations]. AB - This study was aimed to discover the novel HLA recombination haplotypes and investigate the distribution of haplotypes in Chinese Han population. Based on the HLA-A, B, DRB1 typing results of 179 family members, 791 haplotypes were assigned by the mode of inheritance. The results showed that a total of 4 novel recombinant haplotypes in HLA-DRB1 locus region were observed in 4 families, which ratio of paternal to maternal chromosomes was 3:1. The recombination ratio between HLA-DRB1 and HLA-A or B loci was 0.92% (4/433). There were a total of 362 kinds of HLA-A, -B, -DRB1 haplotypes to be confirmed in Chinese Han partial population. A33-B58-DR17, A2-B46-DR9, A30-B13-DR7, A11-B13-DR15, A11-B75-DR12 and A2-B46-DR14 were the most common haplotypes that was consistent with the distribution of HLA alleles in unrelated donors. There were A1-B63-DR12, A29-B46 DR15, A1-B61-DR10, A34-B35-DR9, A29-B54-DR4, A23-B13-DR16 and A34-B62-DR15 haplotypes and so on, which were rare haplotypes not yet reported in Chinese. It is concluded that the HLA-A-B-DRB1 haplotypes would be confirmed by analysis of their family pedigree. The results obtained in this study are basic data for study of Chinese anthropology, organ transplantation and disease correlation analysis. PMID- 19549409 TI - [Investigation on induced expansion of erythroid cells from cord blood CD34(+) cells in vitro]. AB - This study was aimed to investigate a beneficial approach for resolving the deficiency of blood source, preventing the infection resulting from blood transfusion and overcoming the knotty match of patients with rare blood group by using massive expansion of erythroid cells from cord blood CD34(+) cells in vitro. The CD34(+) cells from human cord blood were cultured in serum-free medium supplemented with stem cell factor (SCF), interleukin-3 (IL-3) and erythropoietin (EPO) for 1 week, then expansion and differentiation of CD34(+) cells into erythroid cells were supported by co-culture with human mesenchymal stem cells (MSCs) derived from bone marrow for 2 weeks. The results indicated that after culture for 23 days, the expansion multiple of total cell number reached 2.52 x 10(5), and over 95% of these cells were erythroid cells as compared with less than 1% of myelomonocytic (CD14(+) or CD15(+)) cells and megakaryocytic (CD41(+)) cells. However, the culture system without MSC support was significantly disadvantaged both in expansion ability and ratio of erythroid cells when compared with MSC supporting system. It is concluded that the erythroid cells can be produced from CD34(+) cells in large scale by culturing in the system comprised of cytokine sets and MSC feeders, in which MSCs can support the proliferation and differentiation of erythroid cells. PMID- 19549410 TI - [Application of microfluidic chip analytical systems in ABO genotyping]. AB - Limitations of polyacrylamide gel or agarose gel electrophoretic methods in genotyping research affect the interpreting of detection results. In order to develop a simple and reliable method for appraising results of ABO genotyping detection, the microfluidic chip analysis system was established by using microfluidic chip to replace the gel electrophoresis and combining with multiplex PCR-RFLP technique. 150 blood samples were tested by this microfluidic chip analysis system with multiplex-PCR-RFLP technique to evaluate its stability and accuracy. The results showed that all the testing results were consistent with serologic ABO genotyping results and 1 blood sample with decrease of B antigen caused by CML was identified. In conclusion, the established microfluidic chip analysis system is stable and reliable technique. Application of this technique enables the ABO genotyping results to be more objective and accurate. PMID- 19549411 TI - [Morphology and function of platelets stored in modified platelet additive solution at low temperature]. AB - The aim of study was to evaluate the function of modified platelet additive solution (PAS-IIIM) with trehalose as a substitute of plasma for the storage of platelet concentrates at low temperature (10 degrees C). Apheresis platelets from 6 donors were divided and added with different media (group A: 100% plasma; group B: 70% PAS-IIIM/30% plasma; group C: 100% plasma/trehalose). Groups A, B, C were stored at 10 degrees C, 22 degrees C and -85 degrees C separately. In addition, group D (platelet concentrates stored with 100% plasma at 4 degrees C) was set up as control group for scan electronmicroscopy. The samples of each platelets were collected on day 0, 1, 5, 7 and 9 after storage respectively, while samples of platelets stored at -85 degrees C (group C) were collected on day 20 after storage. CD62p, hypotonic shock response (HSR), platelet aggregation, lactic dehydrogenase (LDH) and morphology of platelets were evaluated. The results showed that the expressions of CD62p in groups A and B increased in a time dependent manner, but HSR and platelet aggregations decreased. The expression of CD62p, LDH release, and platelet aggregation in group A were significant higher than that in group B (p < 0.05). HSR in group A was significant lower than that in group B (p < 0.05). LDH release was significant high in samples of group C and the expression of CD62p was lower than that in other two groups (p < 0.05). It is concluded that the protective effects of 70% PAS-IIIM/30% plasma (10 degrees C) and plasma platelets (22 degrees C) on morphology of platelets are similar, but better than those of plasma platelets (4 degrees C) and plasma/trehalose (-85 degrees C). In short, PAS-IIIM serves as a good substitute of plasma for platelet storage, and protects the chilled platelets. PMID- 19549412 TI - [Evaluation of in vivo viability of human platelets cryopreserved at -80 degrees C by using SCID mouse model]. AB - The purpose of this study was to evaluate the in vivo viability of human platelets cryopreserved at -80 degrees C by using SCID mouse model and flow cytometry. The fresh human platelets were frozen with 5% DMSO at -80 degrees C for 10 days, thawed, and centrifuged for concentration. A 100 ml aliquot of concentrated platelets was injected into the SCID mouse tail vein by using a 1 ml insulin-syringe fitted with a 29-gauge ultra-fine needle. The whole blood was collected into heparinized capillary tube at 0.5, 2, 4, 6, 12, and 24 hours after infusion via a tail vein and was labelled with CD61-PE. Then the human platelets in mouse whole blood were detected by flow cytometry. The 30 minute time point was used as 100% to calculate the survival time of human platelets. The results showed that the survival time of cryopreserved human platelets were more significantly decreased than that of fresh platelets in SCID mice. Survival rates at 4 hours after transfusion of fresh platelets and cryopreserved platelets in SCID mice were 79.5% +/- 9.1% (n = 8) and 40.6% +/- 6.6% (n = 8) respectively, and a T(1/2) estimated were 7 hours for fresh platelets, but 2.5 hours for the cryopreserved. In conclusion, platelets survival time in SCID mice was shortened after frozen with DMSO at -80 degrees C. PMID- 19549413 TI - [Pathogenesis of Fanconi anemia: FA-BRCA network -- review]. AB - Fanconi anemia (FA) is a rare genetic disease with both autosomal and X-linked inheritance, characterized by genomic instability. The cells from individuals with FA are highly sensitive to DNA-crosslinking drugs, such as mitomycin C (MMC), diepoxybutane (DEB) and so on. Now at least 13 genes (FA-A, B, C, D1, D2, E, F, G, I, J, L, M, N) have been identified, whose products participate in a DNA damage-response network involving breast cancer susceptibility gene products, BRCA1 and BRCA2. The impaired DNA repair due to mutations in FA genes is thought to be one of the main pathogenesis of FA, also closely related to the development of some cancers. In this review, the advances of study about FA-BRCA network are summarized. PMID- 19549414 TI - [Research progress on aurora kinase inhibitor MK-0457 in therapy for some hematological malignancies -- review]. AB - Recent work indicates that an Aurora kinase inhibitor MK-0457 (VX-680), a small molecule inhibitor of Aurora kinases A, B, C and BCR-ABL, FLT-3, JAK-2, can block the progression of cell growth cycle, causing apoptosis in a range of human tumors. MK-0457 has the activity against expressions of wild-type and mutated bcr abl gene, including the T315I mutant, and can inhibit the activity of FLT-3, JAK 2 and their mutated types as well. Clinical applications suggest that the MK-0457 has therapeutic effect on the highly refractory CML and CML with poor prognosis, Ph(+) ALL with T315I mutant, relapse refractory AML and JAK-2 positive myeloproliferative diseases (MPD). The intensive preclinical studies and the on going phase II clinical trials will open up a new vista of therapy for some hematological malignancies. This review focuses on the pharmacologic action of MK 0457 and its clinical trial as well as combined application. PMID- 19549415 TI - [Histone modification and its application in therapy for hematologic malignancies -- review]. AB - Histone modification is an important mechanism in oncogenesis and development of hematologic malignancies. Acetylation of lysine residues on histones and opening chromatin are correlated with activation of genes, whereas lysine residues methylation can result in either activation or repression on expressions of chromatin. The main point of all is deacetylation of histone mediated by histone deacetylases (HDACs). HDAC inhibitors are divided into 4 categories: short-chain fatty acids, hydroxamic acids, cyclic tetrapeptides and benzamides, owning different mechanisms in HDAC inhibition. Many kinds of I/II phase clinical tests showed that all these HDAC inhibitors have obviously therapeutic efficacies in treatment of hematologic malignancies with low poisons. Combination of HDAC inhibitors with DNA demethylation drugs can decrease DNA methylation, increase histone acetylation and recover antioncogene expression. As important parts of epigenetics, histone acetylation and HDAC inhibitors possess positive prospects in treatment of hematologic malignancies. In this review the advances of study on mechanisms of histone modification, HDAC inhibitors and their use in treatment of hematologic malignancies are summarized. PMID- 19549417 TI - [Progress of experimental study on myelodysplastic syndrome -- review]. AB - The myelodysplastic syndrome (MDS) is an abnormal clonal proliferation disease resulting from disorder of hematopoietic stem cells/progenitor cells and is characterized by ineffective hematopoiesis and high risk of transforming into acute leukemia. The present experimental studies in gene, chromosome, cytokines and biochemical aspects may put the genetically models for clarifying the pathogenesis of MDS, help to early evaluation of disease prognosis and eventually develop the strategies of more effective prevention and treatment methods for MDS. In this article, the advances of chromosome, gene, cytokines and biochemical aspects in pathogenesis of MDS are summarized on basis of proceedings of ASH meeting in 2007 years. PMID- 19549416 TI - [Dendritic cell-based immunotherapy for multiple myeloma -- review]. AB - Patients with multiple myeloma (MM) have increased constantly in recent years, but treatment for patients with MM is currently unsatisfactory and it is necessary to develop new complementary therapies. Dendritic cells (DCs) are specialized antigen-presenting cells capable of initiating and regulating immune responses. Vaccination with tumor antigen-pulsed DCs has shown to be safe and possesses therapeutic effect against many tumors. In this review, the various types of MM-associated antigens and clinical trials on DC-based immunotherapy in MM are summarized, the development of DC immunotherapy for MM patients in future trials is discussed. PMID- 19549418 TI - [Important role of nitric oxide in stored red blood cells -- review]. AB - The efflux of nitro oxide (NO) in the duration of storing red blood cells (RBCs) was the main reason resulting in decrease and even loss of vasodilatory activity, cell deformability and ability of carrying oxygen (O2) in the stored RBCs. The deep understanding physical functions and acting ways of NO in circulatory system, as well as transformations and balance control of S-Nitrosohemoglobin (SNO-Hb) has an important significance for ensuring sure safety and efficacy of transfusion. In this article, the physical functions, acting ways, retaining and transferring form of nitro oxide, and SNO-Hb adjusting, as well as effects of SNO Hb concentration on change on stored red blood cells were reviewed. PMID- 19549419 TI - [Effect of cluster a in Hox gene on proliferation and differentiation of hematopoietic stem/progenitor cells and its relation to leukemia -- review]. AB - In recent years, Research shows that cluster A of Hox gene family is a group of master genes for proliferation and differentiation of hematopoietic stem/progenitor cells (HSPCs), which influence on the number of hematopoietic stem cells and the differentiation of HSPCs into erythroid, myeloid, megakaryocytic and lymphocytic lineages, and are closely related with the pathogenesis of leukemia. In this review, the effects of gene Hox on proliferation and differentiation of HSPCs were concerned, while the epigenetics alterations of cluster A in Hox gene family as well as its coexistence with Non HOX and other fusion genes were also discussed. This made cluster A in Hox gene family plays a regulatory role in pathogenesis of leukemia. PMID- 19549420 TI - [Research advance on application of platelet-rich plasma in wound repair -- review]. AB - Platelets (Plts) have been shown to play a critical role in tissue repair mechanisms such as chemotaxis, cell proliferation, angiogenesis and extracellular matrix deposition. These effects are largely due to the contents of platelet granules, among which there are a number of important growth factors contributing to the wound repairing process. Platelet-rich plasma (PRP) has been traditionally used as a source of platelet growth factors (PGFs). After connecting activator, platelet gel (PG) is formed and numerous PGFs such as PDGF, TGF, VEGF, IGF, and EGF are released. It is important in different stages of the wound-healing cascade and greatly influence mitogenic and cellular differentiation activities. The aim of this review is to present knowledge about properties of PRP and possibilities of using PRP in the treatment of wound, as well as the success of the clinical studies performed so far, finally the future of PRP therapies are also described. PMID- 19549421 TI - [Factors influencing graft versus leukemia effect -- review]. AB - In treating of leukemia and controlling of minimal-residual disease (MRD), graft versus leukemia effect (GVL) plays a critical role, and complicated mechanisms are involved in this immunology process. When graft cells are infused into recipients, the evoked GVL effect must be inevitably influenced by many factors derived from allogeneic effect between donor and receptor. To utilize GVL more efficiently in future clinical practice and to improve the curative effect of allo-HSCT, it is necessary to recognize these factors. Some potential factors influencing GVL such as chimerism patterns, autocytotoxic cells, dynamics of immune cells in patients, the cytokines and so on are reviewed in this article. PMID- 19549422 TI - Environment, public policy, and human health: implications of current events for the next generation of patients and physicians. PMID- 19549423 TI - The impact of climate change and aeroallergens on children's health. AB - There are unequivocal data that climate change is occurring and that there are resulting health impacts. Climate change can affect the prevalence and severity of allergic and respiratory disorders through projected increases in the temporal and spatial distribution and concentrations of some aeroallergens. This study was designed to critique and summarize existing knowledge on asthma-related impacts of aeroallergen exposure on children in the United States and to provide suggestions about reducing the negative impacts of climate change through increasing education, adapting current management strategies, and modifying distribution channels. A review and synthesis of published literature was performed. Five studies identified evaluated the relationship between aeroallergens and particular symptoms and six evaluated use of the emergency department and hospital care for asthma. Little is known about the relationship between aeroallergens and particular asthma symptoms. However, overall, there appears to be evidence that weed pollen is significantly associated with asthma exacerbations and use of emergency and hospital services. Activities that can help mitigate the impact of additional climate change-induced respiratory disease include continued research, physician and patient education, optimizing production and distribution, and actively considering the budgetary impact of increased prevalence and severity of respiratory disease. Although more research is needed on aeroallergens and respiratory disease, existing studies suggest that it will be essential to consider the health impacts on children. Strategies to reduce the impacts should be developed and implemented now. PMID- 19549424 TI - Increasing insect reactions in Alaska: is this related to changing climate? AB - In 2006, Fairbanks, AK, reported its first cases of fatal anaphylaxis as a result of Hymenoptera stings concurrent with an increase in insect reactions observed throughout the state. This study was designed to determine whether Alaska medical visits for insect reactions have increased. We conducted a retrospective review of three independent patient databases in Alaska to identify trends of patients seeking medical care for adverse reactions after insect-related events. For each database, an insect reaction was defined as a claim for the International Classification of Diseases, Ninth Edition (ICD-9), codes E9053, E906.4, and 989.5. Increases in insect reactions in each region were compared with temperature changes in the same region. Each database revealed a statistically significant trend in patients seeking care for insect reactions. Fairbanks Memorial Hospital Emergency Department reported a fourfold increase in patients in 2006 compared with previous years (1992-2005). The Allergy, Asthma, and Immunology Center of Alaska reported a threefold increase in patients from 1999 to 2002 to 2003 to 2007. A retrospective review of the Alaska Medicaid database from 1999 to 2006 showed increases in medical claims for insect reactions among all regions, with the largest percentage of increases occurring in the most northern areas. Increases in insect reactions in Alaska have occurred after increases in annual and winter temperatures, and these findings may be causally related. PMID- 19549425 TI - Sleep, quality of life, and productivity impact of nasal symptoms in the United States: findings from the Burden of Rhinitis in America survey. AB - Rhinitis is a common chronic condition that has been shown in observational and interventional studies to have a substantial impact on the sufferer. This study was performed to describe the impact of symptoms of allergic rhinitis (AR) on sleep, quality of life, and productivity in a U.S. population. A cohort of AR sufferers and non-AR sufferers was assembled by screening a representative sample of 15,000 households with a self-administered questionnaire in January 2004. A subsample of respondents received a detailed follow-up questionnaire in the May/June pollen season. Of the 7024 individuals with complete data, 3831 met the case definition of AR sufferer; 3193 were non-AR sufferers. Overall, AR sufferers had consistently poorer average scores on the sleep, quality of life, cognition, and productivity scales compared with non-AR sufferers. Subjects with AR symptoms had more sleep impairment (51.2) compared with subjects with non-AR symptoms and those with no symptoms (59.8 and 63.3, respectively). Only 3.6% of subjects with AR symptoms experienced 100% sleep adequacy compared with 11.7% of subjects with non-AR symptoms and 19.2% of subjects with no symptoms. Quality of life and cognition scores were worse in subjects with AR symptoms compared with subjects with non-AR or no symptoms. Work and school productivity was significantly reduced in subjects with AR symptoms in the past 4 weeks compared with subjects with no symptoms (p < 0.05). Individuals who suffer from AR symptoms experience a substantial burden on their ability to sleep, quality of life, cognitive function, and school/workplace productivity. PMID- 19549427 TI - Efficacy and safety of fixed-dose loratadine/montelukast in seasonal allergic rhinitis: effects on nasal congestion. AB - A need exists for safe, effective therapy for the relief of the symptoms of allergic rhinitis (AR) that also consistently relieves nasal congestion, the most common and bothersome symptom. This study was performed to assess efficacy and safety of a once-daily tablet containing 10 mg of loratadine, an antihistamine, and 10 mg of montelukast, a leukotriene antagonist (SCH 445761) versus placebo and pseudoephedrine (PSE; 240 mg once-daily formulation; active comparator). In a multicenter, parallel-group, double-blind, double-dummy, randomized study, 1095 subjects with documented history of seasonal AR and positive skin-prick test to a prevailing aeroallergen were treated for 15 days with fixed-dose combination loratadine/montelukast (L/M), PSE, or placebo. After randomization, subjects rated severity of nasal congestion and measured peak nasal inspiratory flow (PNIF) rate in the morning and evening. The change in quality of life from baseline was also assessed. L/M and PSE were significantly more effective than placebo in alleviating nighttime and daytime nasal congestion and improving PNIF rate, an objective measure of nasal obstruction. There were no significant differences between L/M and PSE for any efficacy analysis including improvement in the quality of life. Subjects treated with L/M experienced a similar incidence of total adverse events versus placebo and a lower incidence of total adverse events (including dizziness, insomnia, jitteriness, nausea, and dry mouth) versus PSE. Nasal decongestant activity of L/M was significantly higher than that of placebo and similar to that of PSE in symptomatic AR subjects. L/M showed a safety profile similar to placebo and was better tolerated than PSE. Thus, L/M offers a safe and efficacious alternative to PSE for the treatment of nasal congestion associated with AR. PMID- 19549426 TI - Comparison of olopatadine 0.6% nasal spray versus fluticasone propionate 50 microg in the treatment of seasonal allergic rhinitis. AB - The efficacy of nasal antihistamines (NAHs) for allergic rhinitis (AR) is comparable with or better than second-generation oral antihistamines, with faster onset of action and greater effect on congestion. Limited data suggest that NAHs may be equivalent to intranasal corticosteroids at reducing the full range of nasal seasonal AR (SAR) symptoms, including congestion. The efficacy of olopatadine 0.6% nasal spray (2 sprays/nostril b.i.d.) for symptoms of SAR was compared with fluticasone 50 microg nasal spray (2 sprays/nostril q.d.) in a double-blind, randomized, parallel-group, 2-week noninferiority trial. A total of 130 symptomatic patients were randomized to treatment and they recorded nasal and ocular allergy symptom scores b.i.d. (morning and evening) in a diary. Both treatments reduced reflective and instantaneous assessments of nasal and ocular symptoms from baseline throughout the 2-week study period (p < 0.05). The reflective total nasal symptom score (the primary efficacy variable) decreased by an average of -45.4% for patients treated with olopatadine 0.6% and by -47.4% for those treated with fluticasone; statistical significance favoring olopatadine was demonstrated at day 1. No significant between-treatment differences were determined for the average 2-week percent changes from baseline for congestion, runny nose, sneezing, itchy nose, and ocular symptoms, although olopatadine had a faster onset of action for reducing all symptoms. Both treatments were safe and well tolerated. Olopatadine and fluticasone nasal sprays both reduced nasal and ocular SAR symptoms with no significant between-treatment differences except for a faster and greater onset of action with olopatadine. PMID- 19549428 TI - Onset of action of loratadine/montelukast in seasonal allergic rhinitis subjects exposed to ragweed pollen in the Environmental Exposure Unit. AB - Onset of action is recognized as an important pharmacologic property of allergic rhinitis (AR) medications. This study was designed to evaluate the onset of action of loratadine/montelukast (L/M; 10 mg/10 mg) versus placebo in subjects with ragweed-induced seasonal AR (SAR). A single-center, double-blind, parallel group study of ragweed-sensitive AR subjects (n = 310) was performed in the Environmental Exposure Unit (EEU). Subjects were exposed to ragweed pollen in the EEU and symptoms were recorded at 30, 60, 90, and 120 minutes before a single dose of L/M or placebo. After dosing, symptoms were recorded for 4 hours, at 15 minute intervals for the first 2 hours and at 30-minute intervals for the final 2 hours. The primary end point was time to onset of action of L/M, defined as the first time point at which the mean change from baseline in total symptom score (TSS) for L/M became and remained significantly better than placebo. Secondary end points included nasal congestion scores and peak nasal inspiratory flow (PNIF). The onset of action of L/M for TSS was 1 hour and 15 minutes (p = 0.005 versus placebo). L/M reduced nasal congestion as indicated by significant improvements in both the nasal congestion score (p = 0.011) and the PNIF measurements (p = 0.007) within 1 hour and 15 minutes postdose. The incidence of treatment-emergent adverse events was similar between groups. The onset of action after treatment with L/M was 1 hour and 15 minutes for TSS, as well as nasal congestion. L/M was well tolerated. PMID- 19549429 TI - Perception of acute airway function changes by patients with mild asthma. AB - The objective of this study was to compare the perception of dyspnea during acute bronchoconstriction and bronchodilation in patients with mild asthma with normal lung function who never experienced severe asthma exacerbations in the past. We studied 83 mild, stable asthmatic patients aged 18-58 years. All patients underwent the methacholine challenge followed by the bronchodilation test with salbutamol. The perceptual sensitivity of changes in airway function was analyzed by linear regression coefficients, slope alpha(constr), and slope alpha(dilat). Additionally, the perception score was determined at a 20% decrease in FEV(1) (PS(20)) during the methacholine challenge. The finding was that perceptual sensitivity for bronchoconstriction and bronchodilation, expressed as slope alpha(constr) and slope alpha(dilat), was similar in the study subjects (mean +/- SD, 0.09 +/- 0.05 and 0.10 +/- 0.05, respectively). The two subgroups under assessment were named poor perceivers when PS(20) < 1 (n = 19) and good perceivers when PS(20) > or = 1 (n = 64). While assessing them, the values of slope alpha(constr) did not differ from the values of slope alpha(dilat) in either of the subgroups of poor perceivers or good perceivers. However, the poor perceivers sensed changes in airway function significantly less than the good perceivers did, although overlapping values of slope alpha were observed. In conclusion, this study indicates that perceptual sensitivity during acute bronchoconstriction and bronchodilation is comparably the same in mild, stable asthmatic patients. However, some of these asthmatic patients may display a diminished perception of dyspnea, which can lead to the deterioration of their asthma without their noticing the corresponding symptoms. Thus, they may delay treatment for acute asthma. PMID- 19549430 TI - Effect of the inhaled corticosteroid mometasone on small airway patency in patients with asthma. AB - The inflammation in asthma involves both the large and the small airways. This study was designed to examine whether mometasone delivered from a dry powder inhaler would improve those parameters thought to reflect patency and obstruction of the small airways (diameter <2 mm). Subjects with mild to moderate asthma, only receiving short-acting beta-agonists, underwent baseline assessment, and then were randomized to receive for 12 weeks either mometasone 400 micrograms once daily in the evening or matching placebo. Outcomes assessed included clinical measures and measures of airway responsiveness and inflammation included methacholine sensitivity (concentration of methacholine causing a 20% fall in forced expiratory volume in 1 second [FEV(1)] [PC(20)]), exhaled nitric oxide, serum ECP, and sputum eosinophils. Pulmonary function was assessed by spirometry, plethysmography, and forced oscillometry. Measures of small airway patency included single breath nitrogen washout and air trapping on expiratory high resolution computed tomography. Results were available on 12 adult subjects who received mometasone and 14 subjects who received placebo. Among tests reflecting small airway patency, the forced expiratory flow between 25 and 75% of vital capacity was significantly improved by mometasone compared with placebo (+9% versus -6%; p = 0.006 and the closing volume over forced vital capacity (FVC; -2% versus 0%; p = 0.05). Other results significantly favoring mometasone over placebo included FEV(1), FVC, PC(20), and A.M. and P.M. peak expiratory flows, and albuterol use. Mometasone delivered by a dry powder inhaler improved asthma control and pulmonary function in tests reflecting both large and small airways. PMID- 19549431 TI - Efficacy and safety evaluation of ciclesonide in mild-to-moderate persistent asthma previously treated with inhaled corticosteroids. AB - Inhaled corticosteroids (ICSs) are recommended as first-line treatment for persistent asthma. This study was designed to evaluate the ability of ciclesonide (CIC) in subjects with stable asthma previously receiving another ICS or ICS/long acting beta(2)-agonist (LABA) to maintain asthma disease control. In this 12 week, multicenter, double-blind, parallel-group study, subjects aged > or =12 years with stable mild-to-moderate persistent asthma were switched at randomization from an ICS or ICS/LABA to CIC, 80 microg twice daily (CIC80 b.i.d.; n = 149); CIC, 160 microg once daily (CIC160 q.d.; n = 150); or placebo (n = 147). Change in forced expiratory volume in 1 second (FEV(1); primary end point), morning peak expiratory flow (PEF), rescue albuterol use, total asthma symptom score, nighttime awakenings, and safety were evaluated. FEV(1) improved from baseline to week 12 after CIC80 b.i.d. treatment (+0.07 L; p = 0.0232), and was maintained after CIC160 q.d. (+0.01 L; p = 0.6217). FEV(1) declined from baseline after placebo (-0.12 L; p < 0.0001) and significantly versus CIC treatments (p < 0.001). At week 12, morning PEF maintained baseline values after CIC80 b.i.d. (-4.43 L/minute; p = 0.1272) and decreased after CIC160 q.d. (-5.77 L/minute; p = 0.0490) and placebo (-12.82 L/minute; p < 0.0001); the difference between CIC80 b.i.d. and placebo was significant (p = 0.035). Baseline albuterol use, total daily asthma score, and nighttime awakenings were maintained after CIC treatments (p > 0.25), but increased after placebo (p < or = 0.002); the difference between CIC80 b.i.d. and placebo was significant (p < 0.02). Incidence of adverse events was similar among treatment groups (range, 52.0-57.9%). In this study, CIC80 b.i.d. maintained asthma control in subjects with stable mild-to moderate asthma previously treated with ICS or ICS/LABA, was well tolerated, and, in general, was better than CIC160 q.d. in maintaining disease control. PMID- 19549432 TI - Efficacy and safety evaluation of ciclesonide in subjects with mild-to-moderate asthma not currently using inhaled corticosteroids. AB - Inhaled corticosteroids (ICSs) are a first-line treatment for persistent asthma. This study was designed to compare the efficacy and safety of ciclesonide (CIC) in subjects with mild-to-moderate persistent asthma not using an ICS. This was a multicenter, double-blind, parallel-group, placebo-controlled, 16-week study in subjects who were > or =12 years old, had a > or =6-month history of persistent asthma, a forced expiratory volume in 1 second (FEV(1)) of > or =60 to < or =85% predicted, and who were not using an ICS < or =30 days before study entry. Subjects were randomized to CIC, 80 microg twice daily (CIC80 b.i.d.; n = 170); CIC, 160 microg once daily in the morning (CIC160 q.d. in the A.M.; n = 173); CIC80 b.i.d. for 4 weeks followed by CIC160 q.d. for 12 weeks (CIC80 b.i.d./CIC160 q.d.; n = 171); or placebo (n = 177). Change in FEV(1) from baseline to the average of weeks 12 and 16 (primary end point) and to week 16, A.M. peak expiratory flow, rescue albuterol use, nighttime awakenings, asthma symptom scores, and safety were evaluated. FEV(1) improved from baseline to the average of weeks 12 and 16 for CIC80 b.i.d. (+0.30L; p < 0.0001), CIC160q.d. (+0.19L; p < 0.0001), CIC80 b.i.d./CIC160 q.d. (+0.19L; p < 0.0001), and placebo (+0.06L; p = 0.0251); improvement was greatest for CIC80 b.i.d. (p < 0.01). At week 16, all CIC treatments significantly improved FEV(1) and A.M. PEF from baseline (p < 0.0001) and compared with placebo (p < or = 0.015). All treatments reduced albuterol use and nighttime awakenings and improved asthma symptom scores (p < or = 0.05 versus baseline); these improvements were greater for CIC80 b.i.d. than for placebo (p < 0.01). The incidence of adverse events was similar among treatment groups (range, 53-58%). In this study, CIC80 b.i.d. improved disease control in subjects with mild-to-moderate persistent asthma not using an ICS and provided greater improvements than CIC160 q.d. PMID- 19549433 TI - Short-term safety of somatropin inhalation powder in adults with mild to moderate asthma. AB - Systemic therapeutic protein delivery through the lungs could potentially replace delivery by injection, but safety needs to be established in patients with known pulmonary disease. This study determined the short-term safety profile of recombinant human growth hormone (rhGH; somatropin) inhalation therapy in clinically stable adult subjects with mild to moderate asthma and methacholine sensitivity. This randomized, placebo-controlled study had two phases: (1) an escalating 3-dose, 4-day/dosage tolerance phase; and (2) a 14-day, crossover design comparability phase. Noninferiority in maintaining forced expiratory volume in 1 second (FEV(1)) was tested for somatropin inhalation powder (SIP) compared with subcutaneously injected rhGH (Hsc) and inhaled placebo. Lung hyperresponsiveness was assessed by methacholine bronchoprovocative challenge, and adverse events (AEs) were recorded. Eight and 18 subjects enrolled in the first and second phases, respectively. Noninferiority of SIP compared with Hsc and placebo was established for FEV(1) after the first and last doses, and noninferiority of SIP compared with Hsc for methacholine challenge was established after the first dose. Pulmonary uptake and systemic distribution of SIP was confirmed by increased serum insulin-like growth factor I levels. Mild, nonprogressive cough and nasal congestion occurred more commonly with SIP. All other treatment-emergent AEs were mild, similar across active treatment groups, and consistent with rhGH treatment. In clinically stable adults with mild to moderate asthma, no significant changes in pulmonary function or worsening of asthma complaints occurred during SIP treatment. Future studies of SIP may enroll subjects with mild to moderate asthma for longer-term evaluation of safety and efficacy. PMID- 19549435 TI - Patients are entitled to maximal IVF pregnancy rates. AB - IVF programmes generally follow treatment protocols that strive for best outcomes. Deviations from such established protocols, even if conceptionally well supported, always risk potentially adverse effects on pregnancy chances. Successful pregnancy represents, however, the principal motivation for all fertility treatments. As a universal medical principle, patients are, therefore, entitled to maximal professional efforts towards their desired outcomes in the safest, quickest and most cost-effective ways. For IVF this means, as multiple patient queries in the literature have demonstrated, the following parameters in this order of importance: highest possible pregnancy rates, lowest possible risks, shortest possible time and lowest possible cost. Some recently widely propagated changes to broadly utilized practice patterns in IVF now, post factum, have been determined to be clinically useless and, in addition, have been shown to adversely affect pregnancy chances. Also post factum, this has led to the acknowledgement that significant modifications to established IVF practice should be introduced with caution. In view of the quite satisfactory IVF pregnancy rates that are currently achieved, the uncontrolled introduction of significant protocol modifications, which may adversely affect IVF outcomes, should no longer be acceptable practice as such unproven practice modifications may violate the patient's entitlement to maximally achievable pregnancy rates. PMID- 19549434 TI - A case of severe refractory chronic urticaria: a novel method for evaluation and treatment. AB - With cholinergic urticaria (ChU), the ultimate diagnosis often depends on the demonstration of characteristic urticaria by appropriate provocation. Several treatment options may be helpful but traditional options (antihistamines, leukotriene inhibitors, and immunosuppressives) may be exhausted by the refractory ChU patient. Here, we describe such a case. Demonstration of immediate hypersensitivity to autologous sweat skin testing (ASwST) may provide a rationale for use of omalizumab (Xolair, Genentech Novartis, South San Francisco, CA). Patients with severe ChU may have difficulty producing sufficient quantities of sweat for ASwST given that the very effort that produces the sample exacerbates ChU. Generation of sweat by iontophoresis with pilocarpine nitrate can be performed at many large medical centers. The procedure is simple, safe, and produces varying amounts of sweat depending on the individual. This sweat can then be used for ASwST. Our patient had a positive ASwST with appropriate positive and negative controls. Our testing methods were validated by negative ASwST, saline control, and positive histamine control in a nonatopic, nonurticarial control patient. By the patient's second injection of omalizumab, her quality of life score was significantly improved, as were her daily medication scores and exercise tolerance. We describe the first case of a patient with severe refractory ChU who had a positive ASwST by a novel collection method who has been successfully treated with omalizumab. We present a novel tool for the evaluation and demonstration of sweat-specific IgE in ChU patients who are unable to provide sweat by more traditional means. PMID- 19549437 TI - A two-tailed Comet assay for assessing DNA damage in spermatozoa. AB - DNA fragmentation is considered an important parameter of semen quality, and of significant value as a predictor of male fertility. Poor quality chromatin is closely associated with, and highly indicative of, some fertility problems. Many methodologies to assess DNA fragmentation in spermatozoa are available, but they are all unable to differentiate between single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in the same sperm cell. The two-tailed Comet assay (2T-Comet) protocol overcomes this limitation. A modification of the original Comet assay was developed for the simultaneous evaluation of DNA SSB and DSB in human spermatozoa. The 2T-Comet assay is a fast, sensitive, and reliable procedure for the quantification and characterization of DNA damage in spermatozoa. It is an innovative method for assessing sperm DNA integrity, which has important implications for human fertility and andrological pathology. PMID- 19549436 TI - Payment of egg donors in stem cell research in the USA. AB - Arguments have been put forth as to whether women who donate oocytes for human embryonic stem cell (HESC) research should be compensated, but data regarding this issue have been scant. Recently in the United States, several States have begun funding HESC research, and patient recruitment efforts have begun. This paper lays out relevant arguments and presents data concerning this issue. Researchers are finding that women are unwilling to donate eggs altruistically, which is hampering the progress of research. These efforts are examined, and data on potential donors' views are presented. The absence of payment, rather than ethical concerns, appears to explain opposition to donation. Women also appear generally ignorant of policies in this area. It is suggested that policy discussions shift focus from whether to pay, to how much would be appropriate, and how to decide; and that research and public and professional education be increased to heighten understanding and awareness of these issues. PMID- 19549438 TI - Seminal plasma total antioxidant capacity and semen parameters in patients with varicocele. AB - Total antioxidant capacity (TAC) was evaluated in the seminal plasma of infertile patients with varicocele in relation to their semen parameters. The study recruited 60 patients affected by varicocele and 10 fertile non-varicocele subjects as controls. Controls had normal semen parameters and proven fertility. On the basis of semen parameters, patients with varicocele were grouped into normozoospermic (n = 12), asthenozoospermic (n = 8), oligoasthenozoospermic (n = 40). The group with oligosthenozoospermia was divided into mild (<20 x 10(6)/ml; > or =15 x 10(6)/ml), moderate (<15 x 10(6)/ml; > or =5 x 10(6)/ml), and severe (<5 x 10(6)/ml), based on sperm count. Antioxidant activity was measured in seminal plasma and peripheral blood using the free oxygen radicals defence test. No significant differences were observed in peripheral blood TAC concentrations between controls and groups. In patients with varicocele and moderate oligoasthenozoospermia or severe oligoasthenozoospermia, seminal plasma TAC concentrations were significantly lower (P < 0.05) than in controls and normozoospermic patients with varicocele. Moreover, in patients with severe oligosthenozoospermia, seminal plasma TAC concentrations were also significantly lower (P < 0.05) than in asthenozoozpermic patients with varicocele. In all subjects, concentrations of TAC showed a positive correlation with sperm concentration (r = 0.93, P < 0.05) and motility (r = 0.92, P < 0.05). PMID- 19549439 TI - Sperm aneuploidies after human recombinant follicle stimulating hormone therapy in infertile males. AB - Errors in sperm chromosome segregation are frequently observed in infertile males. It would therefore be useful to develop methods for reducing the rate of aneuploidy in spermatozoa. Thirty-one males were selected with an elevated frequency of total sperm aneuploidy of sperm chromosomes 18, X and Y by fluorescence in-situ hybridization (FISH): 22 were treated with 150 IU of recombinant FSH on alternate days for 3 months and the other nine (controls) did not receive any hormonal treatment. Before therapy, FISH analysis demonstrated an increased frequency of diploidy (0.663 +/- 0.09%), disomy (0.412 +/- 0.03%) and total aneuploidy (1.30 +/- 0.12%) in the 22 males. Sperm analyses revealed reduced progressive motility (26.73 +/- 2.3%) and a reduced percentage of spermatozoa with normal morphology (23.86 +/- 5.3%). After 90 days of therapy, a significant reduction in aneuploidies (mean total aneuploidy: 0.86% +/- 0.11; P = 0.005) was obtained, as well as an improvement in functional and structural sperm characteristics. In untreated patients, no significant change in semen parameters and frequency of total aneuploidy was observed between baseline (1.054 +/- 0.06%) and 90 days later (1.080 +/- 0.05%). It is therefore suggested that deranged meiotic segregation in spermatozoa could be reduced by FSH treatment. PMID- 19549440 TI - Luteal phase rescue in high-risk OHSS patients by GnRHa triggering in combination with low-dose HCG: a pilot study. AB - Triggering of final oocyte maturation with gonadotrophin-releasing hormone agonist (GnRHa) has previously been shown to prevent ovarian hyperstimulation syndrome (OHSS), but at the same time a detrimental effect on clinical outcome parameters was usually reported. In this prospective, observational, proof-of concept study, a new protocol was employed, using GnRHa to trigger final oocyte maturation in OHSS high-risk IVF/ICSI patients after co-treatment with GnRH antagonist. The aim was to avoid cycle cancellation in high-risk patients without increasing the risk of early onset OHSS and simultaneously secure the reproductive outcome. Twelve patients with >or =25 follicles > or =11 mm in diameter after ovarian stimulation were prospectively enrolled to have final oocyte maturation triggered with 0.5 mg buserelin s.c., followed by a single bolus of 1500 IU human chorionic gonadotrophin (HCG) 35 h later to rescue the luteal phase. A mean of 21.5 oocytes was retrieved. All patients underwent embryo transfer, resulting in an ongoing clinical pregnancy rate per cycle of 50% (6/12) and a live birth rate of 50% (6/12). One patient developed moderate, late onset OHSS that did not require hospitalization. GnRHa triggering of final oocyte maturation followed by one bolus of 1500 IU HCG seems to prevent early onset OHSS in high-risk patients and secure the reproductive outcome. PMID- 19549441 TI - Morphogenesis of polycystic ovaries as assessed by pituitary-ovarian androgenic function. AB - Despite polycystic ovaries (PCO) being a common morphology in women with polycystic ovary syndrome and regular menstruation, the regulatory principles in the morphogenesis of antral follicles have not yet been elucidated. In recognition of the complementary interaction between androgen-induced expression of the FSH receptor and FSH-augmented expression of the androgen receptor in granulose cells of antral follicles, a possible correlation of antral follicle count (AFC) and pituitary-ovarian androgenic function was investigated in 180 infertile women over days 3-5 of the menstrual cycle. Six discrete types of PCO with decreasing pituitary-ovarian androgenic function were identified: Type I (classical Stein-Leventhal syndrome), Type II (hyperandrogenemism), Type III (singular hyper-LH), Type IV (cryptic hyperandrogenism), Type V (relative LH dominancy) and Type VI (relative FSH dominancy), in parallel to a diminishing number of AFC from Type I to Type VI. Because during the early follicular phase of the cycle until the selection of the dominant follicle, antral follicles are composed of newly emerged healthy follicles plus atretic antral follicles that remain non-ovulated from previous cycles, it is proposed that the six types of PCO may represent the folliculogenetic spectra along which PCO morphogenesis proceeds. PMID- 19549442 TI - Pro12Ala and His447His polymorphisms of PPAR-gamma are associated with polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a common endocrine disorder that affects 4 12% of women of reproductive age, and is specified when two of the three following criteria are present: hyperandrogenism, oligomenorrhoea and polycystic ovary morphology by ultrasonography. PCOS is characterized by infertility, obesity and hyperinsulinaemia. However, the aetiology of PCOS has not yet been fully identified due to complex metabolic mechanisms. Many researchers have demonstrated that polymorphisms of putative genes related to symptoms are associated with PCOS. In the reproductive system, energy metabolism and hormonal regulation are important to differentiate granulosa cells in the ovary. Peroxisome proliferator-activated receptor gamma (PPAR-gamma) is a transcription factor, and is known to be associated with insulin sensitivity. Furthermore, it is highly expressed in granulosa cells. It was recently reported that various polymorphisms of PPAR-gamma are associated with PCOS in different ethnic backgrounds. This study has shown that both Pro12Ala and His447His polymorphisms of PPAR-gamma are associated with PCOS in a Korean population. PMID- 19549443 TI - Aromatase gene (CYP19A1) variants, female infertility and ovarian stimulation outcome: a preliminary report. AB - Progress has been made towards ascertaining the genetic predictors of ovarian stimulation in IVF. Aromatase cytochrome P450, encoded by the CYP19A1 gene, catalyses a key step in ovarian oestrogen biosynthesis. Hence, the aromatase gene is an attractive candidate for genetic studies. This study aimed to examine the genetic influences of CYP19A1 TCT trinucleotide insertion/deletion (Ins/Del) and (TTTA)(n) microsatellite intronic polymorphisms on ovarian stimulation outcome and aetiology of female infertility. IVF patients (n = 152) underwent ovarian stimulation according to recombinant FSH and gonadotrophin releasing hormone antagonist protocol. Del/Del homozygous patients with shorter TTTA repeats exhibited decreased ovarian FSH sensitivity in ovarian stimulation, which may reflect variations in aromatase gene expression during early antral follicle development. Accordingly, this study demonstrates correlations between Del allele and shorter (TTTA)(n) repeat sizes with smaller ovaries (r = -0.70, P = 0.047) and fewer antral follicles (r = 0.21, P = 0.018) on days 3-5 of spontaneous menstrual cycle, respectively. Furthermore, Del variation linked with low-repeat number (TTTA)(n) alleles are involved in enhanced genetic susceptibility to unexplained infertility (adjusted OR = 4.33, P = 0.039) and endometriosis (r = 0.88, P = 0.026), which corroborates evidence on the overlapping patient profiles of ovarian dysfunction in both types of female infertility. PMID- 19549444 TI - Automated follicle tracking improves measurement reliability in patients undergoing ovarian stimulation. AB - This study tested the hypothesis that the automated assessment of a stimulated ovary, using 3D ultrasound and sono-AVC (automatic volume calculation), provides quicker analysis of follicular number and size than conventional 2D ultrasound, without any loss in measurement validity. Transvaginal ultrasound was performed on day 10 of stimulation in 89 prospectively recruited subjects undergoing IVF treatment. The number and mean diameter of follicles present in both ovaries was measured manually using 2D ultrasound. 3D data were then acquired and analysed using sono-AVC. Outcome measures included the number of follicles with a mean diameter >9 mm, >13 mm and >17 mm. The time taken for measurements and data acquisition was recorded. The two methods were compared using a paired t-test or the Wilcoxon signed rank test. Complete data were available for 82 subjects. There was no significant difference in the number of follicles with mean diameters >9 mm, >13 mm and >17 mm measured by either method. The total time taken for follicular measurements was significantly less (P < 0.01) for the automated 3D method (180.5 +/- 63.6 versus 236.1 +/- 57.1 s) which was associated with significantly less exposure to ultrasound (39.0 +/- 6.0 versus 236.10 +/- 57.1 s; P < 0.001). Automated 3D follicular measurements using sono-AVC provide a comparable but quicker assessment of follicle number and size. PMID- 19549445 TI - Effect of growth hormone on oocyte competence in patients with multiple IVF failures. AB - In a preliminary, unpublished randomized study conducted in 2000 on 39 patients, including a placebo group, it was observed that the addition of growth hormone (GH) during ovarian stimulation in patients with poor-quality oocytes increased the pregnancy rate. However, the results were not statistically significant due to the small number of patients in each group. A protocol with 8 IU GH was tested in 291 patients with three or more previous failures of embryo transfer for no clearly identifiable reasons. The analysis was restricted to patients receiving either recombinant FSH or human menopausal gonadotrophin (HMG) (n = 245). They were compared retrospectively to all patients with three or more failures during the same period of time but stimulated only with recombinant FSH or HMG, without GH, in an observational study design. Co-stimulation with GH gave better results in terms of number of oocytes collected and embryos obtained. Pregnancy rate per retrieval was higher than in the control group (25.7% versus 18.2%, P < 0.01) and reached a level similar to the one observed in the study centre for the whole population. Ovarian stimulation associated with GH can be proposed for patients with a history of repeated assisted reproduction failures. An improvement of cytoplasmic competence is proposed as an explanation. PMID- 19549446 TI - Oocyte recovery post human follicular fluid centrifugation in modified natural cycle and achieving embryo. AB - This case reports a successful live birth by intracytoplasmic sperm injection (ICSI) following human follicular fluid (HFF) centrifugation for oocyte retrieval in the modified natural cycle of a poor responder patient. A 37-year-old patient presenting with a severe ovarian defect underwent a modified natural cycle with HFF centrifugation prior to ICSI. As there was only one oocyte under direct binocular observation, HFF was centrifuged and a second oocyte was collected. ICSI was performed on both oocytes. Embryo quality and outcome were not compromised by HFF centrifugation. A live birth was achieved in April 2008. In a modified natural cycle, HFF centrifugation avoided loss of oocytes, optimized the IVF treatment, and achieved the development of two embryos. PMID- 19549447 TI - Are there non-invasive markers in human oocytes that can predict pregnancy outcome? AB - Predictive criteria for selection of the best embryo for single embryo transfer remain elusive. This study aimed to determine if non-invasive markers in human oocytes, detectable using polarized light microscopy, can predict pregnancy outcome. Twenty-two pregnancy-producing oocytes from 19 patients had their morphological features compared with 30 oocytes from 19 age-matched patients whose transfer did not result in a pregnancy. Both pregnant and non-pregnant patients had similar numbers of oocytes collected (average: 11.9 +/- 2.8 versus 11.3 +/- 2.9) and similar fertilization rates (70.1% versus 69.6%). All embryos transferred were 4-cell cleavage-stage on day 2 with <10% fragmentation. Meiotic spindles were examined at 39-40 h following human chorionic gonadotrophin administration for spindle normality, length, density and angle from first polar body. There was a significant difference in spindle normality in oocytes in the pregnant patients compared with oocytes in the non-pregnant patients (100% versus 33%, P < 0.001). Spindle density was significantly higher in those oocytes resulting in pregnancy (3.0 +/- 1.23 nm versus 2.5 +/- 0.7 nm, P = 0.02). These oocyte markers may provide a useful non-invasive tool in the selection of the embryo most likely to produce a pregnancy. PMID- 19549448 TI - Zona pellucida birefringence score and meiotic spindle visualization in relation to embryo development and ICSI outcomes. AB - The meiotic spindle and the zona pellucida exhibit molecular order when imaged with polarized optics. This study aimed to investigate possible factors contributing to the zona pellucida birefringence score and meiotic spindle visualization, and to evaluate whether these structures may predict intracytoplasmic sperm injection outcomes. Oocytes were divided into groups according to zona pellucida birefringence and meiotic spindle visualization. In addition, the cycles were split into three groups based on the zona birefringence of transferred embryos. A positive correlation was observed between zona birefringence and meiotic spindle visualization. In addition, when the meiotic spindle was observed, the fertilization rate among oocytes with high or low zona pellucida birefringence was similar. Implantation and pregnancy rates were significantly higher when embryos derived from high birefringence oocytes were exclusively transferred (P = 0.041 and P = 0.004 respectively). Furthermore, the miscarriage rate was higher when embryos derived from low birefringence oocytes were exclusively transferred. On the other hand, the total dose of FSH negatively affected meiotic spindle visualization. Results show that selection of embryos based on zona pellucida and meiotic spindle imaging can significantly improve implantation and pregnancy rates. Moreover, the dose of FSH used for ovarian stimulation may affect the organization of the oocyte meiotic spindle. PMID- 19549449 TI - Prognostic factors for preimplantation genetic screening in repeated pregnancy loss. AB - The objective of this study was to identify specific subgroups of recurrent pregnancy loss (RPL) patients of unknown aetiology in whom the selection of chromosomally normal embryos for transfer improves reproductive outcome in preimplantation genetic screening (PGS). A total of 428 PGS cycles were included and chromosomes 13, 15, 16, 18, 21, 22, X and Y were evaluated. In RPL patients < or =37 years, a lower incidence of chromosomal abnormalities (P = 0.0004) and miscarriages (P = 0.0283) was observed, and there were significantly higher pregnancy (P < 0.0384) and implantation (P < 0.0434) rates than in patients >37 years. In the former subset, results showed: (i) significantly higher implantation rates (P = 0.0411) in couples that had experienced a previous aneuploid miscarriage; (ii) similar aneuploidy, pregnancy and implantation rates in couples suffering previous miscarriages during fertility treatments and in those with previous spontaneous pregnancies; (iii) no miscarriages after PGS in couples in whom a fluorescence in-situ hybridization assay showed the male partner's sperm to be abnormal; and (iv) lower implantation rates in couples with > or =5 previous miscarriages, associated with a lower percentage of chromosomally abnormal embryos. It is concluded that PGS is to be strongly recommended when RPL is associated with miscarriages during infertility treatments, chromosomopathy in a previous miscarriage, up to five previous miscarriages and a high incidence of chromosomal abnormalities in spermatozoa. PMID- 19549450 TI - Anti-Mullerian hormone in pregnant women in relation to other hormones, fetal sex and in circulation of second trimester fetuses. AB - The aim of this study was to investigate the regulation of anti-Mullerian hormone (AMH) blood concentrations in mother and fetus during pregnancy. Serum concentrations of AMH, gonadotrophins, oestradiol and progesterone were measured in pregnant women in the first trimester and AMH concentrations in second trimester fetuses, and these were compared in relation to the sex of the fetus. A total of 153 women undergoing elective termination of a first-trimester pregnancy and seven second-trimester pregnant women undergoing cordocentesis were included. Concentrations of AMH in the serum of first-trimester pregnant women were similar to non-pregnant women and were unrelated to the very high concentrations of human chorionic gonadotrophin and the undetectable concentrations of FSH and LH. Serum concentrations of oestradiol and progesterone were unrelated to the concentrations of AMH and the sex of the fetus. Serum concentrations of AMH of four, second trimester, male fetuses ranged from 64 to 92 ng/ml, whereas it was undetectable in female fetuses. It appears that AMH serum concentrations in first trimester pregnant women seem to be independent of gonadotrophin concentrations and fetal sex. The concentration of AMH in the circulation of male fetuses is higher than previously reported and is a highly sensitive marker for fetal sex. PMID- 19549451 TI - Successful pregnancy following novel IVF protocol and transmyometrial embryo transfer after radical vaginal trachelectomy. AB - Radical vaginal trachelectomy in patients with early-stage cervical cancer is an oncologically safe procedure in well-selected patients. Successful pregnancy in a patient with radical vaginal trachelectomy is possible, with two-thirds of pregnancies resulting in live birth. However, it presents a great challenge for assisted reproductive techniques and reproductive medicine in cases with subsequent severe cervical stenosis. This is a report of a 38-year-old patient who underwent radical vaginal trachelectomy at the age of 33 years for early stage (IA2) adenocarcinoma and subsequently presented with infertility due to cervical factors. The patient underwent ovarian stimulation using a novel SMART (Stimulation with Minimal Adverse effects, Retrieval and Transfer)-IVF protocol. As it was impossible to perform transcervical embryo transfer with an almost absent severely stenotic cervical opening, a transmyometrial embryo transfer under ultrasound guidance was performed. This resulted in a successful singleton full-term pregnancy delivered by Caesarean section at gestational age 37 weeks. As far as is known, this is the first reported case of successful pregnancy conceived by IVF with transmyometrial embryo transfer for a patient who had previously undergone radical vaginal trachelectomy. PMID- 19549452 TI - Human cystic fibrosis embryonic stem cell lines derived on placental mesenchymal stromal cells. AB - This study describes the production of two new human embryonic stem cell (hESC) lines affected by cystic fibrosis. These cell lines are heterozygous compounds, each a carrier of the DF508 mutations associated either with E585X or with 3849+10 kb C-->T. The derivation process was performed on irradiated human placental mesenchymal stromal cells and designed to minimize contact with xeno components. This new source of feeder cells is easy to obtain and devoid of ethical concerns. The cells have a great capacity to proliferate which reduces the need for continuous preparation of new feeder cell lines. In addition, three normal hESC lines were obtained in the same conditions. The five stem cell lines retained hESC-specific features, including an unlimited and undifferentiated proliferation capacity, marker expression and the maintenance of stable karyotype. They also demonstrated pluripotency in vitro, forming cell lineages of the three germ layers, as indicated by immunolocalization of beta-tubulin, alpha fetoprotein and actin. These new genetic cell lines represent an important in vitro tool to study the physiological processes underlying this genetic disease, drug screening, and tissue engineering. PMID- 19549453 TI - Dynamic tests of ovarian reserve: a systematic review of diagnostic accuracy. AB - Despite a plethora of tests of ovarian reserve, there is no perfect test to predict pregnancy. Recent evidence points that anti-Mullerian hormone and antral follicle count may be better than other tests, although other tests continue to be used and form the basis of exclusion of women from fertility treatments. This systematic review concentrated on dynamic tests of ovarian reserve [clomifene citrate challenge test (CCCT), gonadotrophin-releasing hormone agonist stimulation test (GAST) and exogenous FSH ovarian reserve test (EFORT)] and assessed their predictability in terms of fertility outcomes. The study did not restrict itself to women undergoing IVF. The diagnostic odds of abnormal CCCT for non-pregnancy were 2.11 (95% confidence interval, 1.04-4.29) at FSH >10 IU/l (day 3 or 10). The diagnostic accuracy of GAST and EFORT could not be determined due to inconsistencies in the way these tests were conducted. This systematic review and meta-analysis was limited by heterogeneity in terms of the population sampled and the index and reference tests. There is an urgent need for consensus on the performance of these tests and the definition of normality, if their use is to be continued. However, given the present level of evidence, these tests should be completely abandoned. PMID- 19549454 TI - [Aspirin and gastrointestinal bleeding: the problem we confront with]. PMID- 19549455 TI - [Can application of new biological preparations alter the natural course of Crohn's disease]. PMID- 19549456 TI - [Chemoprophylaxis for colorectal cancer: opportunities and challenges]. PMID- 19549457 TI - [Importance of identifying primary aldosteronism among hypertensive population]. PMID- 19549458 TI - [The preventive strategies for cardiovascular complications of diabetics in light of the latest findings from evidence-based medical research]. PMID- 19549462 TI - [A research of migrating motor complex in patients with irritable bowel syndrome]. AB - OBJECTIVES: To compare the migrating motor complex (MMC) in irritable bowel syndrome (IBS) patients with that in healthy controls. To explore whether discrete clustered contractions (DCC) are connected with abdominal pain in IBS patients. To improve the method of measuring gastroenteric motility (esp. jejunum). METHODS: By using 16-channel water-perfused catheter and manometry instruments, MMC in 16 cases of IBS with constipation (IBS-C), 18 cases of IBS with diarrhea (IBS-D) and 18 cases of healthy controls were monitored. RESULTS: The MMC durations of IBS-C and IBS-D patients were (127.5 +/- 25.5) min and (74.5 +/- 18.7) min, respectively. Comparision with those in the control group [(87.5 +/- 24.2) min] showed significant differences (P < 0.001). The contraction amplitudes of stage III in different sites of IBS-C patients decreased significantly as compared with those in the controls [jejunum, (39.8 +/- 11.7) mm Hg vs. (61.1 +/- 14.1) mm Hg, P < 0.001, 1 mm Hg = 0.133 kPa]. The propagation velocities of stage III in different sites of IBS-C patients also decreased significantly as compared with those in the controls [jejunum, (1.8 +/- 0.9) cm/min vs. (2.6 +/- 0.8) cm/min, P < 0.01]. The contraction amplitudes of stage III in different sites of IBS-D patients increased significantly as compared with those in the controls [jejunum, (69.7 +/- 20.5) mm Hg vs. (61.1 +/- 14.1) mm Hg, P < 0.01]. The propagation velocities of stage III in different sites of IBS-D patients also increased significantly as compared with those in the controls [jejunum, (4.1 +/- 2.5) cm/min vs. (2.6 +/- 0.8) cm/min, P < 0.01]. DCC incidences of IBS-C and IBS-D were 87.5% and 88.8%, respectively. Comparision with those in the normal group (83.3%) did not show significant difference (P > 0.05). The prevalences of abnormal stage III contractions (include disturbances and interferences of stage III contractions) in IBS-C and IBS-D patients were 68.8% and 66.7%, respectively; there were no significant differences between the two groups (P > 0.05). However abnormal stage III contractions did not exist in healthy controls. CONCLUSIONS: (1) The MMC of IBS-C and IBS-D patients are changed, as compared with that in healthy people; this implies that small intestinal motility dysfunction is one of the pathogenetic factors of IBS. The abnormal stage III contractions in jejunum may be a predominant change in IBS gastroenteric motility. (2) No apparent connection is found between DCC and pain in IBS. (3) By using 16-channel water-perfused catheter, we first carried out the method of monitoring jejunum contractions in China. Parameters of MMC in Chinese healthy people were investigated, esp. those of jejunum. PMID- 19549463 TI - [The diagnostic value of double-balloon enteroscopy in 67 cases with obscure abdominal pain]. AB - OBJECTIVE: To evaluate the diagnostic valve of double balloon enteroscopy in patients with obscure abdominal pain and analyze the etiology of chronic abdominal pain resulted from enteral diseases. METHODS: Sixty-seven cases with chronic abdominal pain underwent a previous negative gastroscopy, colonoscopy, gastrointestinal barium, B ultrasound and electrocardiogram were received double balloon enteroscopy during June 2005 to June 2008. RESULTS: Thirty-six of 67 patients was done by enteroscopy via anus, and 19 cases via oral, and 12 cases via both anus and oral. The lesions were found in 41 of the 67 patients, with overall diagnostic yield of 61.19%. Among 41 cases of abdominal pain resulted from small bowel diseases, Crohn's disease were found in 15 cases (36.59%), non specific small enteritis in 10 cases (24.39%), tumors in 8 cases (19.51%), other enteral diseases in 8 cases (19.51%). CONCLUSIONS: Double balloon enteroscopy was a diagnostic modality with a high diagnostic value for obscure abdominal pain resulted from small bowel diseases. The most common causes of obscure abdominal pain were Crohn's disease, non-specific small enteritis and tumors. PMID- 19549464 TI - [The relationship of MMP-9, VEGF and PCNA expressions and their clinical significance in gastric adenocarcinoma]. AB - OBJECTIVE: To investigate abnormal protein expression of matrix metalloproteinase 9 (MMP-9), vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in human gastric adenocarcinoma, and further reveal the clinical significance. METHOD: The MMP-9, VEGF and PCNA proteins expression was determined by immunohistochemistry staining in 45 gastric adenocarcinoma tissues, 45 adjacent specimens and 10 normal gastric mucosa tissues via tissue arrays accordingly. The relationship of these protein expression with differentiation degree, development and progression of gastric adenocarcinoma were also analyzed. RESULTS: Positive rates of MMP-9, VEGF and PCNA in gastric adenocarcinoma, adjacent specimens and gastric normal mucosa were as follows: MMP-9, 82.2% (37/45), 64.4% (29/45), 30.0% (3/10) (P = 0.019); VEGF, 73.3% (33/45), 62.2% (28/45), 30.0% (3/10) (P = 0.029); PCNA, 84.4% (38/45), 71.1% (32/45), 10.0% (1/10), there were statistically significant difference (P = 0.001). The positive rates of MMP-9, VEGF and PCNA in well-differentiated adenocarcinoma, moderately differentiated adenocarcinoma and poorly differentiated adenocarcinoma were as follows: MMP-9, 70.0% (7/10), 80.0% (8/10), 88.0% (22/25), there were statistically significant difference (P = 0.015); VEGF, 50.0% (5/10), 60.0% (6/10), 88.0% (22/25), there were statistically significant difference (P = 0.000); PCNA, 60.0% (6/10), 90.0% (9/10), 92.0% (23/25), the difference is significant statistically (P = 0.004). The expression of MMP-9, VEGF and PCNA showed positive relationship with each other by rank correlation analysis (P < 0.05). CONCLUSION: Tissue arrays technology is effective tool to analyze the expression of cancer related proteins in gastric adenocarcinoma. The expression of MMP-9, VEGF and PCNA proteins participates in the tumorigenesis and development process of gastric adenocarcinoma, and these can be used as indexes to evaluate prognosis in clinical. PMID- 19549466 TI - [A comparative study of in-stent restenosis after drug eluting stents]. AB - OBJECTIVE: To compare the long-term effect of three different drug-eluting stents (DES) for in-stent restenotic lesions. METHODS: From April 2004 to June 2006, 390 consecutive patients undergoing DES implantation including 187 Cypher (group C), 89 Taxus (group T) and 114 Firebird (group F) with resulting in-stent restenotic lesions were studied. A mean of 2 year clinical and 7-month angiographic follow up was carried out. RESULTS: Baseline characteristics indicated that there were more unstable angina cases in T group and less left main disease and more triple vessel disease cases in F group. A mean of 2-year follow-up results showed no difference of major adverse cardiac events (MACE) rate among the three groups (23.0% vs 22.5% vs 13.2%, P = 0.081) and no difference of cardiac death, non fatal myocardial infarction (MI) and target vessel revascularization rate (1.1% vs 4.5% vs 1.8%, P = 0.210, 5.9% vs 2.2% vs 2.6%, P = 0.226 and 2.9% vs 2.2% vs 0.9%, P = 0.509). There was no difference of total stent thrombosis or its components among the three groups (total: 4.8% vs 3.4% vs 2.6%, P = 0.605, early: 0.5% vs 0 vs 0.9%, P = 0.560, late: 1.6% vs 1.1% vs 0.9%, P = 0.849 and very late: 2.9% vs 2.2% vs 0.9%, P = 0.509) according to Academic Research Consortium (ARC) standard definitions (definite + probable). 7-month angiographic follow-up indicated that there was a lower trend of both in-stent and in-segment restenosis rate in C and F groups (17.9% vs 29.4% vs 13.6%, P = 0.214 and 21.8% vs 35.3% vs 15.9%, P = 0.132)and in-stent and in-segment late loss was significantly smaller in C and F groups [(0.31 +/- 0.12) mm vs (0.75 +/- 0.24) mm vs (0.31 +/- 0.13) mm, P = 0.000 and (0.33 +/- 0.18) mm vs (0.61 +/- 0.23) mm vs (0.31 +/- 0.14) mm, P = 0.001]. CONCLUSIONS: RESULTS: from this 2-year follow-up, single-center study showed comparable effectiveness and safety of Cypher, Taxus and Firebird DES for in-stent restenotic lesions, but Cypher and Firebird had better effect in reducing restenosis. PMID- 19549465 TI - [A study of HIV-related lipodystrophy syndrome in 55 HIV-infected Chinese adult patients]. AB - OBJECTIVE: To study the prevalence, clinical characteristics and risk factors of HIV-related lipodystrophy syndrome (HIV-LD) in our cohort of HIV-1 infected Chinese adults. METHODS: In a cross-sectional study, 55 HIV-infected patients were recruited from the HIV clinic of Peking Union Medical College Hospital; most of them were undergoing the first-class highly active antiretroviral therapy (HAART) of today in China. Lipoatrophy or lipohypertrophy was defined if there was concordance between the report of fat change and clinical examination of the participants. Whole body dual-energy X-ray absorptiometry (DEXA) scanning was performed. RESULTS: Prevalence of clinical body fat redistribution in the present study was 47.3%. Comparing with non-LD patients, HIV-LD patients had elder age and longer exposure to HAART (P < 0.05). HAART exposure and stavudine (d4T) usage were two independent risk factors for HIV-LD. CONCLUSIONS: HIV-related fat redistribution does exist in Chinese HIV population. Peripheral lipoatrophy occurs commonly in HIV-infected adults but is not associated with increased trunk fat. HAART exposure and especially d4T usage are independent risk factors for HIV LD. PMID- 19549467 TI - [An analysis of the prognostic factors of acute myocardial infarction in different gender]. AB - OBJECTIVE: To analyse the prognostic factors of ST-elevation acute myocardial infarction men and women. METHODS: The data of 904 in-hospital patients with ST elevation myocardial infarction were collected from the database of our hospital during 2003 - 2004 and 728 of them were followed-up. The patients were divided into groups of male and female. RESULTS: Women had more accompanying diseases such as diabetes mellitus (DM) and hypertension than men; left ventricular ejection fraction (LVEF) was lower in female. The rate of successful reperfusion was lower in women than men (P < 0.05). Mortality rate was higher in women. 728 (202 female) patients were followed up. The use of beta-blockers were statistically different between two groups during follow-up. In the female group, LVEF was lower significantly and the rate of readmission for heart failure and myocardial infarction as well as that of mortality was higher (P < 0.05). Multivariate analysis showed that sex difference was an independent risk factor for in-hospital mortality (OR = 2.130, 95% CI 0.954 - 4.754, P = 0.045), but not for mortality in the followed-up period and readmission. CONCLUSION: There are many factors leading to the poor prognosis of ST-elevation acute myocardial infarction in women. It is essential to pay more attention to its clinical characteristics and begin intervention of the risk factors earlier so as to improve the prognosis. PMID- 19549468 TI - [A study on the relationship between metabolic syndrome and anterior cerebral infarction]. AB - OBJECTIVE: To study the relationship between metabolic syndrome (MS) and anterior circulation infarction (ACI). METHODS: 271 ACI patients (166 men and 105 women) who fulfilled the diagnostic criteria of China Guideline for Cerebrovascular Disease Prevention and Treatment were enrolled. 147 control subjects (67 men and 80 women) without the clinical signs of cerebral infarction but with detailed case history, physical examination and CT or MRI were also selected. The prevalence and risk of MS were observed in the ACI and control group. MS was defined with the modified criteria in Chinese. RESULTS: The prevalence of MS in the ACI group and control subjects was respectively 43.17% and 19.05%. The prevalence of MS was significantly higher in the ACI group as compared with the control subjects (P < 0.01). The component level of MS were significantly different between the two groups (P < 0.05). MS was associated with a 3.7 fold higher risk of ACI (P < 0.01). CONCLUSIONS: There is a close relationship between MS and ACI. MS is an important risk factor of ACI. PMID- 19549469 TI - [The results of transbronchial needle aspiration in 164 cases with enlarged mediastinal and/or hilar lymph nodes]. AB - OBJECTIVE: To evaluate the role of transbronchial needle aspiration (TBNA) in the diagnosis of patients with enlarged mediastinal and/or hilar lymph node. METHODS: Patients with mediastinal and/or hilar lymphoadenopathy proven by CT scan were eligible for TBNA as reported. All specimens were directly and instantly smeared for pathological examination. RESULTS: From June 1 2004 to December 31 2007, 164 patients were examined: including 80 lung cancers, 69 lung benign diseases, 2 other malignancy tumor, and 13 without definite diagnosis. Total 260 lymph nodes were punctured. TBNA procedures were successfully carried out in 445/463 (96.1%). Sensitivity of TBNA was 82.5% (66/80) in patients who had been proven to suffer from bronchogenic carcinoma. There were 25 patients that diagnosis of lung cancer was pathologically determined by TBNA only. A total of 122 lymph nodes in the 80 lung cancer patients were aspirated by TBNA with a positive rate of 65.6% (80/122). Severe complications were rare except small amount of bleeding at the TBNA site (100/164, 61.0%). From June 1 2006 to December 31 2007, lymph node tissues able to make histology diagnosis were yield in 73.5% (64/87) patients. Through histology pathology, the sensitivities of TBNA were 53.3% (8/15) for sarcoidosis and 78.6% (33/42) for lung cancer. CONCLUSION: TBNA is quite safe and helpful in diagnosis and staging of bronchogenic carcinoma, and in diagnosis of benign lung diseases. PMID- 19549470 TI - [A clinical analysis 30 cases of lupus mesenteric vasculitis]. AB - OBJECTIVE: To characterize the clinical characteristics of lupus mesenteric vasculitis (LMV). METHODS: Analyzing the clinical, laboratory and treatment data of LMV patients hospitalized from 2002.1.1 to 2007.12.31 retrospectively. RESULTS: (1) The three common manifestations were abdominal pain, diarrhea and vomit with the prevalence rate of 77%, 70% and 67% respectively. (2) The majority of LMV cases were active vital organ (28/30), kidney (24/30) and hematological system (18/30) were the main organs of involvement. Ten patients had hydroureteronephrosis, and 8 patients had intestinal pseudo-obstruction at the same time. (3) Systemic lupus erythematosus disease activity index (SLEDAI) score was > or = 10 in 80% (24/30) of patients. The progression of LMV was accompanied with new-onset leucopenia or worsening leucopenia or hypocomplementemia in 10 cases. (4) Blood antinuclear antibodies were positive in 27 patients detected, and anti-SSA antibody was positive in 15 (56%), anti-U(1)RNP antibody was positive in 14 (52%). (5) Fourteen cases had bowel wall thickening with target sign or mesenteric vessels with palisade or comb sign in contrast CT scan of abdomen. (6) Twenty-seven cases were treated with orally or intravenous medium to high dose steroid therapy and recovered from LMV. CONCLUSIONS: (1) Abdominal pain, diarrhea and vomit were frequent manifestations of LMV patients. (2) LMV was one of the serious complications of systemic lupus erythematosus (SLE), and usually accompanied by active SLE in other organs. (3) A drop in the white blood cell count or complement C(3) titer might be correlate with the occurrence of LMV. It needs to further investigate the relationship between LMV and the high positive rate of anti-SSA and anti-U(1)RNP antibody. (4) LMV patients responded well to intravenous high dose methylprednisolone. PMID- 19549471 TI - [The quantitative assay and clinical significance of JAK2V617F mutation in 131 patients with chronic myeloproliferative disorders]. AB - OBJECTIVE: To investigate the frequency and mutational status of JAK2V617F mutation in Chinese patients with chronic myeloproliferative disorders (CMPD) and to study the relative quantification of mutated JAK2 mRNA and the clinical significance. METHODS: JAK2V617F mutation and the mutational status were screened with amplification-refractory mutation system polymerase chain reaction (ARMS PCR), the relative quantification of mutated JAK2 mRNA was studied by using capillary electrophoresis. RESULTS: A higher prevalence of JAK2V617F in either the heterozygote or homozygote status in essential thrombocythemia (ET) was observed in elderly patients with ET (P < 0.05). The presence of JAK2V617F was found to be significantly correlated with the age at diagnosis (P < 0.05); patients with age > or = 60 years showed significantly higher JAK2 mutated RNA levels than those with age < 60 years (P < 0.05); the presence of JAK2V617F in polycythemia vera (PV) and ET was found to be significantly associated with higher hemoglobin level and higher leukocyte count (P < 0.05). In addition, higher leukocyte count was observed in homozygous ET patients than in heterozygous ET patients (P < 0.05). The frequency of JAK2V617F mutation and the prevalence of homozygote in PV patients were higher than those in ET patients (P < 0.05). The differences of JAK2V617F mRNA levels among PV, ET and chronic idiopathic myelofibrosis (IMF) were not significant. CONCLUSIONS: ARMS-PCR technique can be used to detect the frequency and mutational status of JAK2V617F mutation owing to its sensitivity and along with capillary electrophoresis, quantitative assay for mutated JAK2 mRNA, diagnosis of CMPD and judgement of prognosis become possible. PMID- 19549472 TI - [The effects of glucocorticoid on expression of cannabinoid-1 receptors in osteoclasts from tibial proximal metaphysis of rats]. AB - OBJECTIVE: To explore the molecular mechanism of glucocorticoid (GC)-induced osteoporosis (GIOP). METHODS: Thirty-two female SD rats after matching body weight were divided randomly into three groups: baseline group (n = 10), control group (n = 11) and GC-treated group (n = 11). The administration time was 9 weeks. Bone mineral density (BMD) was measured with dual energy X-ray absorptiometry. A high resolution micro-CT was used to quantify the densitometric and microarchitectural properties of trabeculae in the proximal metaphysis of right tibia. In situ hybridization histochemistry and immunohistochemistry were used to detect the expression of cannabinoid type 1 receptor (CB1R) in the proximal metaphysis of left tibia. RESULTS: At the end of the experiment, whole body BMD in vivo in the control group [(0.156 +/- 0.008) g/cm(2)] was higher than that in the baseline group [(0.147 +/- 0.006) g/cm(2)], while the whole-body BMD in vivo [(0.147 +/- 0.006) g/cm(2)] and total BMD in vitro at femurs in the GC treated group [(0.220 +/- 0.011) g/cm(2)] was lower than those in the control group [(0.240 +/- 0.024) g/cm(2)]. Compared with the baseline group and control group, there was a remarkable decrease in the volumetric BMD, tissue BMD, trabecular number and trabecular connectivity (P < 0.05) in the GC-treated group, while there was a significant increase in trabecular separation (P < 0.05) and trabecular thickness also increased in the proximal metaphysis of tibiae in the GC-treated group. The expression level of CB1R mRNA and protein in osteoclasts in the GC-treated group was markedly higher than that in the baseline group and control group (P < 0.05). There was a close correlation between the expression level of CB1R mRNA, protein in osteoclasts and some microarchitectural parameters in the proximal metaphysis in the GC-treated group (P < 0.05). CONCLUSIONS: The administration of GC is associated with a decrease in BMD and deterioration in microarchitecture of trabecular bone in rats tibiae. Glucocorticoid may up regulate the CB1R expression level in osteoclasts and this may be a kind of molecular mechanism of GIOP. PMID- 19549486 TI - Modulation of the immunogenicity of the Trypanosoma congolense cysteine protease, congopain, through complexation with alpha(2)-macroglobulin. AB - The protozoan parasite Trypanosoma congolense is the main causative agent of livestock trypanosomosis. Congopain, the major lysosomal cysteine proteinase of T. congolense, contributes to disease pathogenesis, and antibody-mediated inhibition of this enzyme may contribute to mechanisms of trypanotolerance. The potential of different adjuvants to facilitate the production of antibodies that would inhibit congopain activity was evaluated in the present study. Rabbits were immunised with the recombinant catalytic domain of congopain (C2), either without adjuvant, with Freund's adjuvant or complexed with bovine or rabbit alpha(2) macroglobulin (alpha(2)M). The antibodies were assessed for inhibition of congopain activity. Rabbits immunised with C2 alone produced barely detectable anti-C2 antibody levels and these antibodies had no effect on recombinant C2 or native congopain activity. Rabbits immunised with C2 and Freund's adjuvant produced the highest levels of anti-C2 antibodies. These antibodies either inhibited C2 and native congopain activity to a small degree, or enhanced their activity, depending on time of production after initial immunisation. Rabbits receiving C2-alpha(2)M complexes produced moderate levels of anti-C2 antibodies and these antibodies consistently showed the best inhibition of C2 and native congopain activity of all the antibodies, with maximum inhibition of 65%. Results of this study suggest that antibodies inhibiting congopain activity could be raised in livestock with a congopain catalytic domain-alpha(2)M complex. This approach improves the effectiveness of the antigen as an anti-disease vaccine candidate for African trypanosomosis. PMID- 19549487 TI - Expression of three intelectins in sheep and response to a Th2 environment. AB - Sheep intelectin1 and sheep intelectin3 (sITLN1 and sITLN3) were cloned and sequenced. The amino acid sequences of sITLN1 and sITLN3 shared 86% and 91% homology with the previously cloned sheep intelectin2 (sITLN2), respectively. Expression of sITLN1 and sITLN3 transcript was demonstrated in abomasum, lung, colon and gastric lymph node, terminal rectum, skin, jejunum, mesenteric lymph node, ileal peyer's patches, brain, kidney, liver, spleen, skin, ear pinna, heart and ovary in normal sheep tissues. sITLN2 transcript expression was restricted to the abomasal mucosa in normal sheep tissues. Using a non selective chicken anti intelectin antibody, tissue intelectin protein was demonstrated in mucus neck cells in the abomasum, mucus cells in the colon, free mucus in ileum, goblet cells in the lung, small intestinal epithelium and brush border, epidermal layer of the skin and skin sebaceous glands. The expression of the three sITLN transcripts was examined in two nematode infections in sheep known to induce a Th2 response; a Teladorsagia circumcincta challenge infection model and a Dictyocaulus filaria natural infection. The three sITLN were absent in unchallenged naive lambs and present in the abomasal mucosa of both naive and immune lambs following T. circumcincta challenge infection. Upregulation of sITLN2 and sITLN3 was shown in sheep lung following D. filaria natural infection. Intelectins may play an important role in the mucosal response to nematode infections in ruminants. PMID- 19549489 TI - The BBSome. PMID- 19549488 TI - Bovine neonate natural killer cells are fully functional and highly responsive to interleukin-15 and to NKp46 receptor stimulation. AB - Natural killer (NK) cells are key components of the innate immune system with their killing and cytokine producing abilities. Bovine NK cells have been characterized as NKp46(+)/CD3(-) lymphocytes, but little is known about these cells in neonatal calves. As the newborn calf, with an insufficiently developed acquired immunity, has to employ the innate immune system, we wanted to investigate whether neonate NK cells had the same characteristics as cells from older calves. Freshly isolated neonate and calf NK cells presented the same resting CD2(+)/CD25(low)/CD8(-/low) phenotype. Neonates less than 8 days old had one third of the circulating NKp46(+) cells of older calves, but the NK cells proliferated more actively in vitro in the presence of interleukin (IL)-2 or IL 15. Moreover, neonate NK cells were more cytotoxic both in an NKp46 mediated redirected lysis assay and in direct killing of a bovine cell line MDBK when cultured in the presence of IL-15. Neonate and calf NK cells cultured in the presence of IL-2 and then stimulated with IL-12 produced similar dose-dependent interferon (IFN)-gamma amounts, while IL-15 cultured NK cells did not give such a response whatever the age. However, neonatal NK cells cultured in IL-15 and stimulated by IL-12 concomitantly with cross-linking of NKp46, produced 4 to 5 times more IFN-gamma than calf NK cells. These data suggest that although present in lower number at birth, neonate NK cells are fully functional and are more responsive to IL-15 and activation through the NKp46 receptor than NK cells from older calves. PMID- 19549490 TI - Animal asymmetry. PMID- 19549491 TI - Tool-use induces morphological updating of the body schema. PMID- 19549492 TI - Sleep: what goes up must come down. AB - The function of sleep is hotly contested. Two recent studies suggest that fly sleep may be required to rescale synapses in the brain. PMID- 19549493 TI - Chromosome segregation: monopolin goes spindle. AB - At anaphase onset the mitotic spindle undergoes dramatic changes in order to segregate sister chromatids. Surprisingly, the monopolin complex, best known for its role at kinetochores in meiosis, is now shown to localize to, and stabilize, the mitotic anaphase spindle. PMID- 19549494 TI - Learning and memory: while you rest, your brain keeps working. AB - A recent study shows that brain activity recorded while the human subject is at 'rest' is significantly affected by a prior learning episode. These results suggest that understanding resting brain activity may be critical to understanding how humans learn from experience. PMID- 19549495 TI - Cognitive ecology: environmental dependence of the fitness costs of learning. AB - A recent study has found that butterflies maintain behavioural plasticity useful to them in rare environments by reducing associated costs in common environments. Butterflies use innate sensory biases to locate common green hosts, but learn to modify these preferences to find rare, red host-plants. PMID- 19549496 TI - Oogenesis: making the mos of meiosis. AB - Meiosis is an ancient type of cell division whose advent allowed the evolution of sexual reproduction. The evolutionary history of the specialization that allowed gamete production to emerge from a simple reduction division has been unclear. New data now suggest that the molecular mechanisms involved in animal oocyte specialization may have origins that predate the emergence of bilaterian animals. PMID- 19549498 TI - Visual perception: saccadic omission--suppression or temporal masking? AB - Although we don't perceive visual stimuli during saccadic eye movements, new evidence shows that our brains do process these stimuli and they can influence our subsequent visual perception. PMID- 19549497 TI - Olfaction: chemical signposts along the silk road. AB - A recent study on the reception of olfactory cues by silkworm larvae illustrates how the convergence of genomic, physiological and ecological data promises to shed light on the origins and evolution of chemically mediated interactions between plants and insects. PMID- 19549499 TI - Antigenic variation: extending the reach of telomeric silencing. AB - Immune evasion in the parasitic African trypanosome relies upon the silencing of variant surface glycoprotein genes that are found adjacent to telomeres. Work on the RAP1 telomere-binding protein now indicates that silencing spreads over a sufficient distance to repress these genes. PMID- 19549500 TI - Protein phosphorylation influences proteolytic cleavage and kinase substrate properties exemplified by analysis of in vitro phosphorylated Plasmodium falciparum glideosome-associated protein 45 by nano-ultra performance liquid chromatography-tandem mass spectrometry. AB - Plasmodium falciparum glideosome-associated protein 45 (PfGAP45) was in vitro phosphorylated by P. falciparum calcium-dependent protein kinase (PfCDPK1) and digested using the four proteases trypsin, chymotrypsin, AspN, and elastase. Subsequently, phosphopeptide enrichment using Ga(III) immobilized metal affinity chromatography (IMAC) was performed. The resulting fractions were analyzed using ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS), resulting in the identification of a total of nine phosphorylation sites: Ser31, Ser89, Ser103, Ser109, Ser121, Ser149, Ser156, Thr158, and Ser173. During in-depth analyses of the detected phosphopeptides, it was observed that phosphorylation alters the properties of PfGAP45 as kinase and protease substrate. The closely adjacent phosphorylation sites Ser156 (major site) and Thr158 (minor site) were analyzed in detail because at first glance the specific proteases gave highly variable results with respect to the relative abundance of these sites. It was observed that (i) formation of pSer156 and pThr158 was mutually exclusive and (ii) phosphorylation at Ser156 or Thr158 interfered specifically with proteolysis by chymotrypsin or trypsin, respectively. The latter effect was studied in detail using synthetic phosphopeptides carrying either pSer156 or pThr158 as substrate for chymotrypsin or trypsin, respectively. PMID- 19549501 TI - Effect of storage regime on the stability of DNA used as a calibration standard for real-time polymerase chain reaction. AB - This article looks at storage factors influencing the stability of potential DNA calibration standards for use in quantitative polymerase chain reaction (PCR). Target sequences from the bacteria Campylobacter jejuni were cloned into a plasmid vector. Samples of these potential calibration standards were stored at +4, -20, and -80 degrees C as aqueous and lyophilized samples and were prepared as both single-use aliquots and multiple-use preparations. Results showed that the samples stored as single-use aqueous solutions at +4 degrees C and lyophilized samples stored at +4 and -20 degrees C were the most stable. Samples stored as frozen aqueous solutions at -20 degrees C were the least stable. PMID- 19549502 TI - Nitrogen oxide air pollutants interfere with the measurement of nitric oxide using 2,3-diaminonaphthalene: reduction of background interference. AB - Nitrite, a stable metabolite of nitric oxide (NO), is measured fluorometrically as an indicator of NO production using 2,3-diaminonaphthalene. In cultured cells, it has been believed that a longer period of incubation improves the detection sensitivity because of the accumulation of nitrite formed from NO in culture media. However, here we show that nitrite formed from nitrogen oxide air pollutants accumulates continuously in culture media during the incubation and interferes with the measurement of NO as nitrite. Thus, a proper period of incubation is important to allow maximum nitrite signals from NO with minimum background nitrite from the air. PMID- 19549503 TI - Mapping of HIV-1 integrase preferences for target site selection with various oligonucleotides. AB - HIV integrase (IN) catalyzes the insertion of proviral DNA into the host cell chromosome. While IN has strict sequence requirements for the viral cDNA ends, the integration site preference has been shown to be very diverse. Here, we mapped the HIV IN strand transfer reaction requirements using various short oligonucleotides (ON) that mimic the target DNA. Most double stranded DNA dodecamers served as excellent IN targets with variable integration efficiency depending mostly on the ON sequences. The preferred integration was lost with any changes in the geometry of the DNA double helical structures. Various hairpin loop-forming ONs also served as efficient integration targets. Similar integration preferences were also observed for ONs, in which the nucleotide hairpin loop was replaced with a flexible aliphatic linker. The integration biases with all target DNA structures tested were significantly influenced by changes in the resulting secondary ON structures. PMID- 19549504 TI - Reconstitution of blue fluorescent protein from recombinant apoaequorin and synthetic coelenteramide. AB - Blue fluorescent protein of aequorin (BFP) is a complex of Ca(2+)-bound apoaequorin with coelenteramide and is a bifunctional protein, which shows blue fluorescence and the luminescence activity like a luciferase. To reconstitute synthetic BFP (syn-BFP) from apoaequorin and coelenteramide, we established new synthetic route of coelenteramide and prepared highly purified recombinant aequorin using the histidine-tagged secretion system in Escherichia coli cells. As a result, we succeeded in reconstituting syn-BFP quantitatively and the fluorescence and luminescence properties of syn-BFP were identical to that of BFP obtained from aequorin. PMID- 19549505 TI - Interview with Aaron Ciechanover. PMID- 19549506 TI - Comparative pharmacology and computational modelling yield insights into allosteric modulation of human alpha7 nicotinic acetylcholine receptors. AB - The human alpha7 nicotinic acetylcholine receptor (nAChR) subunit and its Caenorhabditis elegans homolog, ACR-16, can generate functional recombinant homomeric receptors when expressed in Xenopus laevis oocytes. Both nAChRs express robustly in the presence of the co-injected chaperone, RIC-3, and show striking differences in the actions of a type I positive allosteric modulator (PAM), ivermectin (IVM). Type I PAMs are characterised by an increase in amplitude only of the response to acetylcholine (ACh), whereas type II PAMs exhibit, in addition, changes in time-course/desensitization of the ACh response. The type I PAMs, ivermectin, 5-hydroxyindole (5-HI), NS-1738 and genistein and the type II PAM, PNU-120596, are all active on human alpha7 but are without PAM activity on ACR-16, where they attenuate the amplitude of the ACh response. We used the published structure of avermectin B1a to generate a model of IVM, which was then docked into the candidate transmembrane allosteric binding site on alpha7 and ACR 16 in an attempt to gain insights into the observed differences in IVM actions. The new pharmacological findings and computational approaches being developed may inform the design of novel PAM drugs targeting major neurological disorders. PMID- 19549507 TI - Long term treatment with ACE inhibitor enalapril decreases body weight gain and increases life span in rats. AB - Renin-angiotensin system is involved in homeostasis processes linked to renal and cardiovascular system and recently has been linked to metabolic syndrome. We analyzed the influence of long term angiotensin I converting enzyme (ACE) inhibitor enalapril treatment in normotensive adult Wistar rats fed with standard or palatable hyperlipidic diets. Our results show that long term enalapril treatment decreases absolute food intake, serum leptin concentration and body weight gain. Moreover, in adipose tissue, enalapril treatment led to decreased ACE activity, enhanced the expression of peroxisome proliferator activated receptor gamma, adiponectin, hormone-sensitive lipase, fatty acid synthase, catalase and superoxide dismutase resulting in prolonged life span. On the other hand, the ACE inhibitor was not able to improve the transport of leptin through the blood brain barrier or to alter the sensitivity of this hormone in the central nervous system. The effect of enalapril in decreasing body weight gain was also observed in older rats. In summary, these results extend our previous findings and corroborate data from the literature regarding the beneficial metabolic effects of enalapril and show for the first time that this ACE inhibitor prolongs life span in rats also fed with palatable hyperlipidic diet, an action probably correlated with adipose tissue metabolic modulation and body weight reduction. PMID- 19549510 TI - Dimebolin is a 5-HT6 antagonist with acute cognition enhancing activities. AB - Dimebolin (Dimebon), is a non-selective antihistamine approved in Russia for the treatment of allergy. Recently, this drug has been shown to be neuroprotective in cellular models of Alzheimer's disease and Huntington's disease, and to preserve cognitive function when chronically administered to AF64A lesioned rats. Interests in identifying the molecular targets of dimebolin have intensified with reports of efficacy in clinical trials with Alzheimer's patients. Dimebolin has been found to interact with a number of molecular targets including acetylcholinesterases, N-methyl-d-aspartate receptors, and voltage-gated calcium channels, with potencies in the range of 5-50 microM. In the present study, the action of dimebolin at the serotonin 5-HT(6) receptor was investigated. Dimebolin binds with moderate affinity to both the human and rat recombinant 5-HT(6) receptor (K(i)=26.0+/-2.5 nM and 119.0+/-14.0 nM respectively) as well as the native rat 5-HT(6) receptor, and acts as an antagonist in functional cAMP assays. Furthermore, dimebolin occupies the 5-HT(6) receptor in vivo as assessed by ex vivo autoradiography, with a dose-occupancy relationship similar to that of the selective 5-HT(6) antagonist SB-399885. Finally, both SB-399885 and dimebolin produce an acute enhancement of short-term social recognition memory, although dimebolin is approximately 10-fold less potent than SB-399885. Taken together, these studies demonstrate that dimebolin antagonizes the 5-HT(6) receptor with higher affinity than other targets characterized to date, and suggest that this activity may play a role in the acute cognition enhancing effects of this compound in preclinical models and in the clinic. PMID- 19549508 TI - PCB-induced endothelial cell dysfunction: role of poly(ADP-ribose) polymerase. AB - Polychlorinated biphenyls (PCBs) are persistent environmental pollutants implicated in the development of pro-inflammatory events critical in the pathology of atherosclerosis and cardiovascular disease. PCB exposure of endothelial cells results in increased cellular oxidative stress, activation of stress and inflammatory pathways leading to increased expression of cytokines and adhesion molecules and ultimately cell death, all of which can lead to development of atherosclerosis. To date no studies have been performed to examine the direct effects of PCB exposure on the vasculature relaxant response which if impaired may predispose individuals to hypertension, an additional risk factor for atherosclerosis. Overactivation of the DNA repair enzyme poly(ADP-ribose) polymerase (PARP) following oxidative/nitrosative stress in endothelial cells and subsequent depletion of NADPH has been identified as a central mediator of cellular dysfunction. The aim therefore was to investigate whether 2,2',4,6,6' pentachlorobiphenyl (PCB 104) directly causes endothelial cell dysfunction via increased oxidative stress and subsequent overactivation of PARP. Exposure of ex vivo rat aortic rings to PCB 104 impaired the acetylcholine-mediated relaxant response, an effect that was dependent on both concentration and exposure time. In vitro exposure of mouse endothelial cells to PCB 104 resulted in increased cellular oxidative stress through activation of the cytochrome p450 enzyme CYP1A1 with subsequent overactivation of PARP and NADPH depletion. Pharmacological inhibition of CYP1A1 or PARP protected against the PCB 104-mediated endothelial cell dysfunction. In conclusion, the environmental contaminants, PCBs, can activate PARP directly impairing endothelial cell function that may predispose exposed individuals to development of hypertension and cardiovascular disease. PMID- 19549511 TI - Pediatric reference intervals determined in ambulatory and hospitalized children and juveniles. AB - OBJECTIVES: The aim of this study was to determine reference intervals in children and juveniles with nine recently developed and widely used laboratory methods. METHODS: More than 800 ambulatory and hospitalized individuals of the University pediatry were carefully selected according to clinical status and chemical profile in an a posteriori process over a period of two and a half years. The reference group with well-balanced gender proportions and steady age distribution between 1 day and 17 years was subdivided in five age classes. The laboratory methods were: the enzyme methods ALT, AST, LDH and GGT, all reliably reference standardized with traceability to the IFCC reference methods at 37 degrees C; ALT and AST without pyridoxal-phosphate activation; ALP as not yet approved IFCC method; the soluble transferrin receptor (sTfR) and ferritin, the latter being the only heterogeneous procedure. RESULTS: The results confirm in most cases the typical age concentration relationship of the measured quantities documented for similar methods. In some critical cases, in particular for sTfR and ferritin, the study produces limits which differ distinctly from those earlier reported. Gender differentiation was outlined according to statistical calculations. CONCLUSION: Proposals for reference intervals were derived from the central 95 percentile values. PMID- 19549509 TI - Retinoid metabolism and ALDH1A2 (RALDH2) expression are altered in the transgenic adenocarcinoma mouse prostate model. AB - Retinoids, which include vitamin A (retinol) and metabolites such as retinoic acid, can inhibit tumor growth and reverse carcinogenesis in animal models of prostate cancer. We analyzed retinoid signaling and metabolism in the TRAMP (transgenic adenocarcinoma mouse prostate) model. We detected increased retinol and retinyl esters in prostates pooled from 24 to 36 week TRAMP transgenic positive mice compared to nontransgenic littermates by HPLC. We used quantitative RT-PCR to measure transcripts for genes involved in retinoid signaling and metabolism, including ALDH1A1, ALDH1A2, ALDH1A3, CYP26A1, LRAT, and RARbeta(2), in prostate tissue from TRAMP positive (+) and age-matched littermate control mice ranging from 18 to 36 weeks. Transcript levels of ALDH1A1, a putative stem cell marker, were decreased in ventral and lateral lobes of prostates from TRAMP mice compared to age-matched, nontransgenic mice. ALDH1A2 (RALDH2) mRNA levels in dorsal and anterior lobes of TRAMP+ mice were lower than in age-matched (24 week) nontransgenic mice. We detected lower RARbeta(2) mRNA levels in dorsal prostate lobes of 36 week TRAMP mice relative to nontransgenic mice. We detected high levels of ALDH1A2 protein in the cytoplasm and nucleus in nontransgenic murine prostate paraffin sections, and lower ALDH1A2 protein levels in all prostate lobes of TRAMP mice compared to nontransgenic mice by immunohistochemistry. We also detected much lower cytoplasmic ALDH1A2 protein levels in all human prostate cancer paraffin sections stained (19 total) relative to normal human prostate tissue on the same sections. Our data indicate that this reduction in ALDH1A2 protein is an early event in human prostate cancer. PMID- 19549512 TI - Protective effects of kaempferol against endothelial damage by an improvement in nitric oxide production and a decrease in asymmetric dimethylarginine level. AB - Previous investigations have shown that asymmetric dimethylarginine (ADMA) inhibits nitric oxide (NO) synthases (NOS) and that ADMA is a risk factor for endothelial dysfunction. The objective of this study was to investigate the protective effect of kaempferol, a naturally occurring flavonoid antioxidant agent, against endothelial damage and the mechanisms involved. The experiments were performed in aorta and plasma from C57BL/6J control and apolipoprotein E deficient (ApoE(-/-)) mice treated or not with kaempferol (50 or 100mg/kg, intragastrically) for 4 weeks, and in human umbilical vein endothelial cells (HUVECs) pretreated or not with kaempferol (1, 3 or 10 microM) for 1h and exposed to lysophosphatidylcholine (LPC) (10 microg/mL) for 24h. Kaempferol treatment improved endothelium-dependent vasorelaxation, increased the maximal relaxation value, and decreased the half-maximum effective concentration concomitantly with an increase in nitric oxide plasma concentration, a decrease in ADMA and malondialdehyde (MDA) plasma concentrations, and increase in the expression of aortic endothelial NOS (eNOS) as well as dimethylarginine dimethylaminohydrolase II (DDAH II) in ApoE(-/-) mice. In addition, LPC caused a reduction in NO production, an increase in ADMA concentration concomitantly with a decreased expression of eNOS and DDAH II in HUVECs, and the effect of LPC was abolished by kaempferol. Treatment with kaempferol also significantly decreased reactive oxygen species production in mice aorta and in HUVECs. The present results suggest that the protective effect of kaempferol against endothelial damage may be associated with an improvement in NO production and a decrease in ADMA level. PMID- 19549513 TI - Milrinone inhibits hypoxia or hydrogen dioxide-induced persistent sodium current in ventricular myocytes. AB - Much evidence indicates that increased persistent sodium current (I(Na.P)) is associated with cellular calcium overload and I(Na.P) is considered to be a potential target for therapeutic intervention in ischaemia and heart failure. By inhibiting type III phosphodiesterase, milrinone increases intracellular cyclic adenosine monophosphate (cAMP), with a positive inotropic effect. However, the effect of milrinone on increased I(Na.P) under pathological conditions remains unknown. Accordingly, we investigated the effect of milrinone on increased I(Na.P) induced by hypoxia or hydrogen dioxide in guinea pig ventricular myocytes. While milrinone (0.01 mM or 0.1mM) or cAMP (0.1 mM) decreased I(Na.P) respectively in control condition, application of 1 microM H-89, a selective cAMP dependant protein kinase inhibitor, prevented the effect of 0.1mM milrinone in control condition. Milrinone (0.1 mM) reduced the increased I(Na.P) induced by hypoxia. Furthermore, 0.01 mM or 0.1mM milrinone reduced the enhanced I(Na.P) induced by 0.3 mM hydrogen peroxide. In addition, 0.01 mM or 0.1 mM milrinone shortened action potential duration at 90% repolarization (APD(90)). Bath application of 0.3 mM hydrogen dioxide markedly prolonged APD(90), while 2 microM tetrodotoxin (TTX) reversed the prolonged APD(90). In the other two groups, 0.01 mM or 0.1 mM milrinone shortened the prolonged APD(90) induced by 0.3 mM hydrogen peroxide, ultimately 2 microM TTX causing a further decurtation of APD(90). These findings demonstrate that milrinone inhibited I(Na.P) under normal condition, hypoxia or hydrogen dioxide-induced I(Na.P), and the APD(90) prolonged by hydrogen dioxide-induced I(Na.P) in ventricular myocytes, which is associated with the mechanism of milrinone increasing intracellular cAMP. PMID- 19549514 TI - Antidepressant effects of pramipexole, a dopamine D3/D2 receptor agonist, and 7 OH-DPAT, a dopamine D3 receptor agonist, in olfactory bulbectomized rats. AB - Treatment with pramipexole, a dopamine D(3)/D(2) receptor agonist, reduces depressive symptoms in patients suffering from Parkinson's disease. To test the putative antidepressant quality of pramipexole, its effects were assessed in one of the most attractive animal models of depression, the olfactory bulbectomized (OBX) rat. Two experiments studied the effects of pramipexole on bulbectomy induced hyperactivity. In experiment I, pramipexole was tested at 0.3 and 1.0 mg/kg together with the reference dopamine D(3) receptor agonist 7-OH-DPAT (0.1 mg/kg) and the tri-cyclic antidepressant imipramine (10 mg/kg). In experiment II, pramipexole was tested at lower doses: 0.03 and 0.1 mg/kg, with the same reference compounds. All animals were tested in the open field on days one (acute), seven (sub-chronic) and fourteen (chronic) of administration, as well as one week after cessation of treatment. Pramipexole, in a U-shaped dose response, reduced bulbectomy-induced hyperactivity after (sub) chronic but not acute administration (like imipramine and 7-OH-DPAT). The highest dose of pramipexole (1.0 mg/kg) did not reduce OBX hyperactivity during treatment. However, one week after cessation of treatment, all pramipexole (including the 1.0 mg/kg dose), 7 OH-DPAT and imipramine groups showed a reduction in OBX-induced hyperactivity. Pramipexole and 7-OH-DPAT exert an antidepressant profile in the OBX-rat model in normalizing bulbectomy-induced hyperactivity during (sub) chronic treatment. Moreover, treatment with both these compounds induced long-lasting changes in the bulbectomized brain similar to established antidepressants, strongly predicting antidepressant activity in major depression. PMID- 19549515 TI - Inhibitory effect of lidocaine on pain and itch using formalin-induced nociception and 5'-guanidinonaltrindole-induced scratching models in mice: behavioral and neuroanatomical evidence. AB - The aim of this study was to establish the effect of lidocaine, a local anesthetic, on pain and itch using formalin-induced nociception and kappa opioid antagonist-induced scratching models in mice. We investigated if local intradermal pretreatment (at -10 min) with lidocaine N-ethyl bromide (lidocaine, 2%, 0.1 ml) antagonizes behavioral responses and prevents c-fos expression induced by pain and itch. Male, Swiss Webster mice (25-30 g, n=6-10) were used. Formalin (5%, 20 microl, s.c.) or saline was administered to the right dorsal hindpaw and the time spent licking this paw was recorded at 0-10 min and 20-35 min. For itching, mice were challenged with 5'-guanidinonaltrindole (GNTI, 0.3mg/kg, s.c., behind the neck) or saline and the number of neck-directed scratches with hindpaws was counted for 30 min. C-fos immunohistochemistry was performed in lumbar (for pain) and cervical (for scratching) spinal sections 2h after the respective treatments. We found that lidocaine (a) antagonizes both formalin-induced pain and GNTI-induced scratching and (b) prevents c-fos expression evoked by pain (medial side of the superficial layer and deeper layers of the dorsal horn) and itch (lateral side of the superficial layer of the dorsal horn). Additionally, GNTI caused c-fos activation in mice wearing an Elizabethan collar (to prevent scratching of the neck) suggesting that GNTI provokes c-fos expression by inducing an itch sensation. Our results highlight the antipruritic properties of lidocaine and argue for its comprehensive clinical testing against pruritic states. PMID- 19549516 TI - Apigenin protects endothelium-dependent relaxation of rat aorta against oxidative stress. AB - Apigenin is shown to have cardiovascular effects, but the effects of apigenin on aortas injured by exogenous oxidants are unknown. The objective of this study was to investigate the effect of apigenin on endothelium-dependent vasorelaxation in isolated rat aortic rings exposed to superoxide anion produced by pyrogallol, and its mechanism. The male Sprague-Dawley rat thoracic aorta was rapidly dissected out and the effect of apigenin on tension of aortic rings pretreated with 500 microM pyrogallol, inducing oxidative stress injury, was measured. The activity of nitric oxide synthase (NOS), the level of nitric oxide (NO) and the inhibition of superoxide anion in aortic tissues were measured. We found that pretreatment with pyrogallol concentration-dependently decreased acetylcholine-induced endothelium-dependent vasorelaxation. Apigenin (0.5-72.0 microM) evoked a concentration-dependent relaxation in aortas (pD(2): 5.304+/-0.049), which was weakened by L-NAME (the maximal relaxation fell from 87.6+/-6.7% to 37.1+/-8.8%, P<0.01), but not by aminoguanidine and indomethacin. Apigenin markedly attenuated the inhibition of vasorelaxation induced by pyrogallol (the maximal relaxation elevated from 55.8%+/-6.6% to 69.5%+/-6.4%, and the pD(2) increased from 6.559+/ 0.119 to 7.057+/-0.145, P<0.01) and increased the inhibition of superoxide anion (from 94.6% to 74.5%), the NO level (from 77.1% to 94.4%), and the constitutive NOS activity (from 35.1% to 62.5%). These results indicate that pyrogallol decreased endothelium-dependent vasorelaxation in rat aortas via oxidative stress, which was markedly attenuated by apigenin. This may be mediated by weakening the oxidative stress and the NO reduction. PMID- 19549518 TI - Use of latency to immobility improves detection of antidepressant-like activity in the behavioral despair test in the mouse. AB - The behavioral despair test (BDT), also called the forced swim test, is an economic, reliable and sensitive test for the detection of potential antidepressant-like activity of new test substances. The vast majority of clinically active antidepressants are active in the BDT, although substances specifically acting on serotonin transmission are generally reported to be less easily detected. Substances active in the BDT decrease the duration of immobility at doses considered as relatively high. In contrast, some psychostimulants are considered as potential false positives since they are also active in the BDT although they are not recognized as clinically active antidepressants. In the present study we have evaluated the usefulness of latency to the first immobility period as an additional parameter in the BDT to further evaluate the effects of antidepressants and psychostimulants administered intraperitoneally in the mouse. The results show that this measure increases the sensitivity of the test for detecting the effects of tricyclic antidepressants (imipramine, desipramine) and selective serotonin/norepinephrine reuptake inhibitors (duloxetine and venlafaxine) but not of serotonin reuptake inhibitors (fluoxetine and escitalopram). In contrast with previous reports, psychostimulants (amphetamine and modafinil) did not affect the duration or the latency to immobility in the BDT. The mouse strain used in the BDT seems to be an important parameter to discriminate between antidepressants and psychostimulants. These results suggest that the measure of the latency to the first immobility improves the predictive validity of the BDT. PMID- 19549517 TI - Two allelic isoforms of the serotonin transporter from Schistosoma mansoni display electrogenic transport and high selectivity for serotonin. AB - The human blood fluke Schistosoma mansoni is the primary cause of schistosomiasis, a debilitating disease that affects 200 million individuals in over 70 countries. The biogenic amine serotonin is essential for the survival of the parasite and serotonergic proteins are potential novel drug targets for treating schistosomiasis. Here we characterize two novel serotonin transporter gene transcripts, SmSERT-A and SmSERT-B, from S.mansoni. Southern blot analysis shows that the two mRNAs are the products of different alleles of a single SmSERT gene locus. The two SmSERT forms differ in three amino acid positions near the N terminus of the protein. Both SmSERTs are expressed in the adult form and in the sporocyst form (infected snails) of the parasite, but are absent from all other stages of the parasite's complex life cycle. Heterologous expression of the two cDNAs in mammalian cells resulted in saturable, sodium-dependent serotonin transport activity with an apparent affinity for serotonin comparable to that of the human serotonin transporter. Although the two SmSERTs are pharmacologically indistinguishable from each other, efflux experiments reveal notably higher substrate selectivity for serotonin compared with their mammalian counterparts. Several well-established substrates for human SERT including (+/-)MDMA, S (+)amphetamine, RU 24969, and m-CPP are not transported by SmSERTs, underscoring the higher selectivity of the schistosomal isoforms. Voltage-clamp recordings of SmSERT substrate-elicited currents confirm the substrate selectivity observed in efflux experiments and suggest that it may be possible to exploit the electrogenic nature of SmSERT to screen for compounds that target the parasite in vivo. PMID- 19549519 TI - Effect of oral mucosal adhesive films containing ginsenoside Rb1 on 5 fluorouracil-induced oral mucositis in hamsters. AB - Oral ulcerative mucositis induced by chemotherapy or radiotherapy has an impact on quality of life, is dose-limiting for chemotherapy, and causes considerable morbidity. The aim of this study was to evaluate the effect of ginsenoside Rb1 on 5-fluorouracil (5-FU)-induced experimental oral mucositis in hamsters. Oral mucositis was induced in hamsters through a combination of 5-FU treatment and mild abrasion of the cheek pouch. Ginsenoside Rb1 isolated from ginseng was contained in chitosan-sodium alginate film (G-Rb1 film). The films were attached to the oral mucosa, and then the healing process was examined by measuring the area of mucositis, myeloperoxidase (MPO) activity and microscopic aspects. Films without ginsenoside Rb1 had no effect on 5-FU-induced oral mucositis in comparison to the control group. However, G-Rb1 films (10(-12) to 10(-4) g/g film) dose-dependently improved recovery from 5-FU-induced damage, and there were significant differences between doses of 10(-6) and 10(-4) g/g film. These results suggest that topical application of films that contain ginsenoside Rb1 has a healing effect on severe oral mucositis induced by chemotherapy. PMID- 19549520 TI - AtCYP20-2 is an auxiliary protein of the chloroplast NAD(P)H dehydrogenase complex. AB - AtCYP20-2 is one of 16 immunophilins in thylakoid lumen. The presence of the isomerase domain in AtCYP20-2, an enrichment of AtCYP20-2 in the stroma membranes and it's co-migration with NAD(P)H dehydrogenase (NDH) in native gels provide evidence that AtCYP20-2 is an auxiliary protein of NDH. When different NDH mutants were studied, AtCYP20-2 was found to be strongly reduced especially in mutants deficient in the membrane domain of NDH, thus suggesting a role in the assembly of NDH hydrophobic domain. Lack of AtCYP20-2, however, did not lead to severe malfunction of NDH, indicating redundancy in the function of lumenal immunophilins. PMID- 19549521 TI - Temperature-dependent hyper-activation of monoglucosyldiacylglycerol synthase is post-translationally regulated in Synechocystis sp. PCC 6803. AB - The mechanism of monoglucosyldiacylglycerol (MGlcDG) increase following heat shock in Synechocystis sp. PCC 6803 was examined by measuring MGlcDG synthase (Sll1377) activity. Temperature-dependent activation of Sll1377 was observed in the membrane fraction of Synechocystis sp. PCC 6803, whereas the Sll1377 protein level remained unchanged, suggesting that the activity is post-translationally regulated without covalent modification of Sll1377 by soluble enzymes. Four individual mutations introduced into recombinant Sll1377 (D147, D200, R329, and R331) significantly reduced the activity and blocked temperature-dependent activation, suggesting that these amino acid residues are essential for Sll1377 activity at both normal growth temperature and the higher temperature. PMID- 19549522 TI - c-Jun N-terminal kinase-1 from hematopoietic cells mediates progression from hepatic steatosis to steatohepatitis and fibrosis in mice. AB - BACKGROUND & AIMS: c-Jun N-terminal kinase (JNK) plays a pivotal role in the development of the metabolic syndrome including nonalcoholic fatty liver disease. However, the mechanism underlying the contribution of JNK to the progression from simple steatosis to steatohepatitis and liver fibrosis is unresolved. METHODS: Hepatic steatosis, inflammation, and fibrosis were examined in wild-type, jnk1(-/ ), or jnk2(-/-) mice fed a choline-deficient L-amino acid-defined (CDAA) diet for 20 weeks. The functional contribution of JNK isoforms in Kupffer cells was assessed in vitro and in vivo using chimeric mice in which the hematopoietic compartment including Kupffer cells was replaced by wild-type, jnk1(-/-), or jnk2(-/-) cells. RESULTS: CDAA diet induced significantly less hepatic inflammation and less liver fibrosis despite a similar level of hepatic steatosis in jnk1(-/-) mice as compared with wild-type or jnk2(-/-) mice. CDAA diet-induced hepatic inflammation was chronic and mediated by Kupffer cells. Pharmacologic inhibition of JNK or gene deletion of jnk1 but not jnk2 repressed the expression of inflammatory and fibrogenic mediators in primary Kupffer cells. In vivo, CDAA diet induced less hepatic inflammation and liver fibrosis despite an equivalent level of hepatic steatosis in chimeric mice with jnk1(-/-) hematopoietic cells as compared with chimeric mice with wild-type or jnk2(-/-) hematopoietic cells. CONCLUSIONS: jnk1(-/-) mice are resistant to diet-induced steatohepatitis and liver fibrosis. JNK1 in hematopoietic cells, especially in Kupffer cells, contributes to the development of liver fibrosis by inducing chronic inflammation. PMID- 19549524 TI - Peripheral and central P2X receptor contributions to colon mechanosensitivity and hypersensitivity in the mouse. AB - BACKGROUND & AIMS: Irritable bowel syndrome is characterized by altered sensory qualities, namely discomfort/pain and colorectal hypersensitivity. In mice, we examined the role of P2X(3) receptors in colon mechanosensitivity and intracolonic zymosan-produced hypersensitivity, a model of persistent colon hypersensitivity without colon inflammation. METHODS: The visceromotor response to colon distension (15-60 mm Hg) was determined before and after intracolonic saline or zymosan (30 mg/mL, 0.1 mL, daily for 3 days) treatment. Colon pathology and intracolonic adenosine triphosphate release was assessed in parallel experiments. To examine P2X(3) receptor contributions to colon mechanosensation and hypersensitivity, electrophysiologic experiments were performed using an in vitro colon-pelvic nerve preparation. RESULTS: Visceromotor responses to distension were significantly reduced in P2X(3)(+/-) and P2X(3)(-/-) mice relative to wild-type mice. Colon hypersensitivity produced by zymosan was virtually absent in P2X(3)(-/-) relative to wild-type or P2X(3)(+/-) mice. Intralumenal release of the endogenous P2X receptor ligand adenosine triphosphate did not differ between wild-type and P2X(3)(-/-) mice or change after intracolonic zymosan treatment. Responses of muscular and muscular-mucosal pelvic nerve afferents to mechanical stretch did not differ between P2X(3)(-/-) and wild type mice. Both muscular and muscular-mucosal afferents in wild-type mice sensitized to application of an inflammatory soup, whereas only muscular-mucosal afferents did so in P2X(3)(-/-) mice. CONCLUSIONS: These results suggest differential roles for peripheral and central P2X(3) receptors in colon mechanosensory transduction and hypersensitivity. PMID- 19549525 TI - Deletion of TRPC4 and TRPC6 in mice impairs smooth muscle contraction and intestinal motility in vivo. AB - BACKGROUND & AIMS: Downstream effects of muscarinic receptor stimulation in intestinal smooth muscle include contraction and intestinal transit. We thought to determine whether classic transient receptor potential (TRPC) channels integrate the intracellular signaling cascades evoked by the stimulated receptors and thereby contribute to the control of the membrane potential, Ca-influx, and cell responses. METHODS: We created trpc4-, trpc6-, and trpc4/trpc6-gene deficient mice and analyzed them for intestinal smooth muscle function in vitro and in vivo. RESULTS: In intestinal smooth muscle cells TRPC4 forms a 55 pS cation channel and underlies more than 80% of the muscarinic receptor-induced cation current (mI(CAT)). The residual mI(CAT) depends on the expression of TRPC6, indicating that TRPC6 and TRPC4 determine mI(CAT) channel activity independent of other channel subunits. In TRPC4-deficient ileal myocytes the carbachol-induced membrane depolarizations are diminished greatly and the atropine-sensitive contraction elicited by acetylcholine release from excitatory motor neurons is reduced greatly. Additional deletion of TRPC6 aggravates these effects. Intestinal transit is slowed down in mice lacking TRPC4 and TRPC6. CONCLUSIONS: In intestinal smooth muscle cells TRPC4 and TRPC6 channels are gated by muscarinic receptors and are responsible for mI(CAT). They couple muscarinic receptors to depolarization of intestinal smooth muscle cells and voltage activated Ca(2+)-influx and contraction, and thereby accelerate small intestinal motility in vivo. PMID- 19549523 TI - Coinfection with HIV-1 and HCV--a one-two punch. AB - Hepatitis C virus (HCV) infection is a major cause of chronic liver disease, cirrhosis, and death; it is estimated that 180 million persons are infected with HCV worldwide. The consequences of HCV are worse in those who are coinfected with human immunodeficiency virus 1 (HIV-1), which is unfortunately a common scenario because of shared risk factors of the viruses. More studies into effects of HCV/HIV-1 coinfection are needed, but efforts have been hampered by limitations in our understanding of the combined pathogenesis of the 2 viruses. Gaining insight into the mechanisms that underlie the immunopathogenesis of these persistent viral infections could lead to new therapeutic strategies for patients with HCV/HIV-1 coinfection. PMID- 19549527 TI - The ets-domain transcription factor Spdef promotes maturation of goblet and paneth cells in the intestinal epithelium. AB - BACKGROUND & AIMS: Stem cells within the intestinal epithelium generate daughter cells that undergo lineage commitment and maturation through the combined action of the Wnt and Notch signaling cascades. Both pathways, in turn, regulate transcription factor networks that further define differentiation toward either enterocytes or 1 of 3 secretory cell lineages (Paneth, goblet, or enteroendocrine cells). In this study, we investigated the role of the Wnt-responsive, Ets-domain transcription factor Spdef in the differentiation of goblet and Paneth cells. METHODS: The in vivo function of Spdef was examined by disrupting the Spdef gene in mice (Spdef(-/-) mice) and analyzing the intestinal phenotype using a range of histologic techniques and DNA microarray profiling. RESULTS: In accordance with expression data, we found that loss of Spdef severely impaired the maturation of goblet and Paneth cells and, conversely, led to an accumulation of immature secretory progenitors. Spdef appears to positively and negatively regulate a specific subset of goblet and Paneth cell genes, including Cryptdins, Mmp7, Ang4, Kallikreins, and Muc2. CONCLUSIONS: Spdef acts downstream of Math1 to promote terminal differentiation of a secretory progenitor pool into Paneth and goblet cells. PMID- 19549526 TI - Pathogenic bacteria induce colonic PepT1 expression: an implication in host defense response. AB - BACKGROUND & AIMS: Expression of the di/tripeptide transporter PepT1 has been observed in the colon under inflammatory conditions; however, the inducing factors and underlying mechanisms remain unknown. Here, we address the effects of pathogenic bacteria on colonic PepT1 expression together with its functional consequences. METHODS: Human colonic HT29-Cl.19A cells were infected with the attaching and effacing enteropathogenic Escherichia coli (EPEC). Wild-type and PepT1 transgenic mice or cultured colonic tissues derived from these mice were infected with Citrobacter rodentium, a murine attaching and effacing pathogen related to EPEC. RESULTS: EPEC induced PepT1 expression and activity in HT29 Cl.19A cells by intimately attaching to host cells through lipid rafts. Induction of PepT1 expression by EPEC required the transcription factor Cdx2. PepT1 expression reduced binding of EPEC to lipid rafts, as well as activation of nuclear factor-kappaB and mitogen-activated protein kinase and production of interleukin-8. Accordingly, ex vivo and in vivo experiments revealed that C rodentium induced colonic PepT1 expression and that, compared with their wild type counterparts, PepT1 transgenic mice infected with C rodentium exhibited decreased bacterial colonization, production of proinflammatory cytokines, and neutrophil infiltration into the colon. CONCLUSIONS: Our findings demonstrate a molecular mechanism underlying the regulation of colonic PepT1 expression under pathologic conditions and reveal a novel role for PepT1 in host defense via its capacity to modulate bacterial-epithelial interactions and intestinal inflammation. PMID- 19549528 TI - Endoscopist-directed administration of propofol: a worldwide safety experience. AB - BACKGROUND & AIMS: Endoscopist-directed propofol sedation (EDP) remains controversial. We sought to update the safety experience of EDP and estimate the cost of using anesthesia specialists for endoscopic sedation. METHODS: We reviewed all published work using EDP. We contacted all endoscopists performing EDP for endoscopy that we were aware of to obtain their safety experience. These complications were available in all patients: endotracheal intubations, permanent neurologic injuries, and death. RESULTS: A total of 646,080 (223,656 published and 422,424 unpublished) EDP cases were identified. Endotracheal intubations, permanent neurologic injuries, and deaths were 11, 0, and 4, respectively. Deaths occurred in 2 patients with pancreatic cancer, a severely handicapped patient with mental retardation, and a patient with severe cardiomyopathy. The overall number of cases requiring mask ventilation was 489 (0.1%) of 569,220 cases with data available. For sites specifying mask ventilation risk by procedure type, 185 (0.1%) of 185,245 patients and 20 (0.01%) of 142,863 patients required mask ventilation during their esophagogastroduodenoscopy or colonoscopy, respectively (P < .001). The estimated cost per life-year saved to substitute anesthesia specialists in these cases, assuming they would have prevented all deaths, was $5.3 million. CONCLUSIONS: EDP thus far has a lower mortality rate than that in published data on endoscopist-delivered benzodiazepines and opioids and a comparable rate to that in published data on general anesthesia by anesthesiologists. In the cases described here, use of anesthesia specialists to deliver propofol would have had high costs relative to any potential benefit. PMID- 19549529 TI - Cyclooxygenase-2 inhibitors down-regulate osteopontin and Nr4A2-new therapeutic targets for colorectal cancers. AB - BACKGROUND & AIMS: Cyclooxygenase-2 inhibitors reduce colon cancer risk by mechanisms that are not fully understood. We performed microarray analysis of adenomas from Apc(Delta14/+) mice to identify genes that respond to these drugs. METHODS: Apc(Delta14/+) mice were given a single daily injection of parecoxib for up to 9 weeks; intestinal tracts of these and control mice were analyzed by microarray analysis, immunohistochemistry, in situ hybridization, and quantitative real-time polymerase chain reaction. Findings were further assessed using Apc(lox/lox)vil-CreER(T2) mice, the CT26 cancer cell line, and human colon tumor samples. RESULTS: Microarray analysis revealed that osteopontin, a marker of colon cancer progression, was down-regulated in polyps from Apc(Delta14/+) mice given parecoxib compared with controls. Apc(Delta14/+) mice given parecoxib had longer survival times and reduced polyp burdens. Osteopontin was quickly down regulated by parecoxib in intestinal polyps from Apc(Delta14/+) mice, and 2 components of the osteopontin regulatory network-the orphan nuclear receptor NR4A2 and Wnt/beta-catenin signaling-were sequentially repressed. NR4A2 activated the osteopontin promoter in CT26 cells; this effect was blocked by mutation of the NR4A2 binding response element, cotransfection of a dominant-negative form of NR4A2, and small inhibitory RNA against NR4A2. NR4A2 levels were increased throughout tumor progression in Apc(Delta14/+) mice but, unlike osteopontin, did not correlate with tumor stage. NR4A2 levels were reduced in adenomas from patients treated with rofecoxib. CONCLUSIONS: Down-regulation of osteopontin, probably through blockade of NR4A2 and Wnt signaling, is an important component of the antitumor activity of cyclooxygenase-2 inhibitors. These factors might be developed as therapeutic targets for intestinal cancers. PMID- 19549530 TI - Induction and down-regulation of Sox17 and its possible roles during the course of gastrointestinal tumorigenesis. AB - BACKGROUND & AIMS: The activation of Wnt/beta-catenin signaling causes the development of gastric and colon cancers. Sox17 represses Wnt/beta-catenin signaling and is down-regulated in colon cancer. This study was designed to elucidate the role of Sox17 during the course of gastrointestinal tumorigenesis. METHODS: Sox17 expression was examined in gastrointestinal tumors of mouse models and humans. The roles of Sox17 in gastric tumorigenesis were examined by cell culture experiments and by construction of Sox17 transgenic mice. RESULTS: Sox17 was induced in K19-Wnt1/C2mE mouse gastric tumors and K19-Wnt1 preneoplastic lesions, where Wnt/beta-catenin signaling was activated. Consistently, Wnt activation induced Sox17 expression in gastric cancer cells. In contrast, Sox17 was rarely detected by immunohistochemistry in gastric and colon cancers, whereas strong nuclear staining of Sox17 was found in >70% of benign gastric and intestinal tumors. Treatment with a demethylating agent induced Sox17 expression in gastric cancer cells, thus indicating the down-regulation of Sox17 by methylation. Moreover, transfection of Sox17 in gastric cancer cells suppressed both the Wnt activity and colony formation efficiency. Finally, transgenic expression of Sox17 suppressed dysplastic tumor development in K19-Wnt1/C2mE mouse stomach. CONCLUSIONS: Sox17 plays a tumor suppressor role through suppression of Wnt signaling. However, Sox17 is induced by Wnt activation in the early stage of gastrointestinal tumorigenesis, and Sox17 is down-regulated by methylation during malignant progression. It is therefore conceivable that Sox17 protects benign tumors from malignant progression at an early stage of tumorigenesis, and down-regulation of Sox17 contributes to malignant progression through promotion of Wnt activity. PMID- 19549531 TI - Expansion and differentiation of neural progenitors derived from the human adult enteric nervous system. AB - BACKGROUND & AIMS: Neural stem and progenitor cells from the enteric nervous system have been proposed for use in cell-based therapies against specific neurogastrointestinal disorders. Recently, enteric neural progenitors were generated from human neonatal and early postnatal (until 5 years after birth) gastrointestinal tract tissues. We investigated the proliferation and differentiation of enteric nervous system progenitors isolated from human adult gastrointestinal tract. METHODS: Human enteric spheroids were generated from adult small and large intestine tissues and then expanded and differentiated, depending on the applied cell culture conditions. For implantation studies, spheres were grafted into fetal slice cultures and embryonic aganglionic hindgut explants from mice. Differentiating enteric neural progenitors were characterized by 5-bromo-2-deoxyuridine labeling, in situ hybridization, immunocytochemistry, quantitative real-time polymerase chain reaction, and electrophysiological studies. RESULTS: The yield of human neurosphere-like bodies was increased by culture in conditional medium derived from fetal mouse enteric progenitors. We were able to generate proliferating enterospheres from adult human small or large intestine tissues; these enterospheres could be subcultured and maintained for several weeks in vitro. Spheroid-derived cells could be differentiated into a variety of neuronal subtypes and glial cells with characteristics of the enteric nervous system. Experiments involving implantation into organotypic intestinal cultures showed the differentiation capacity of neural progenitors in a 3 dimensional environment. CONCLUSIONS: It is feasible to isolate and expand enteric progenitor cells from human adult tissue. These findings offer new strategies for enteric stem cell research and future cell-based therapies. PMID- 19549532 TI - Coordination of deiodinase and thyroid hormone receptor expression during the larval to juvenile transition in sea bream (Sparus aurata, Linnaeus). AB - To test the hypothesis that THs play an important role in the larval to juvenile transition in the marine teleost model, sea bream (Sparus auratus), key elements of the thyroid axis were analysed during development. Specific RT-PCR and Taqman quantitative RT-PCR were established and used to measure sea bream iodothyronine deiodinases and thyroid hormone receptor (TR) genes, respectively. Expression of deiodinases genes (D1 and D2) which encode enzymes producing T3, TRs and T4 levels start to increase at 20-30 days post-hatch (dph; beginning of metamorphosis), peak at about 45 dph (climax) and decline to early larval levels after 90-100 dph (end of metamorphosis) when fish are fully formed juveniles. The profile of these different TH elements during sea bream development is strikingly similar to that observed during the TH driven metamorphosis of flatfish and suggests that THs play an analogous role in the larval to juvenile transition in this species and probably also in other pelagic teleosts. However, the effect of T3 treatment on deiodinases and TR transcript abundance in sea bream is not as clear cut as in larval flatfish and tadpoles indicating divergence in the responsiveness of TH axis elements and highlighting the need for further studies of this axis during development of fish. PMID- 19549535 TI - Cocaine and automaintained responding in pigeons: rate-reducing effects and tolerance thereto with different durations of food delivery. AB - Pigeons were exposed to an automaintenance procedure in which 6-s key illuminations in one color (red or white) were immediately followed by 3-s food deliveries and key illuminations in the other color were followed by 9-s food deliveries. Both conditions engendered consistent responding. With both durations of food delivery, acute and chronic cocaine administrations (1.0-17.8 mg/kg) produced dose-dependent decreases in mean percent trials (key illuminations) with a response and mean total response per session. Tolerance developed to the disruptive effects of cocaine on both response measures. Food duration did not significantly affect either response measure or significantly interact with cocaine dose or drug regimen. The orderliness of the present findings, like those of a related study examining whether probability of food delivery modulated the effects of cocaine on automaintained responding [Porritt, M., Arnold, M., Poling, A., Cocaine and automaintained responding in pigeons: rate-reducing effects and tolerance thereto with different CS-US pairing probabilities. Pharmacol Biochem Behav 2007; 87:405-411.], suggests that the automaintenance procedure is a useful assay for examining tolerance to drug effects on classically-conditioned responding. Unlike the results of that study, however, the present findings are inconsistent with a behavioral momentum analysis of drug effects on such responding. PMID- 19549534 TI - Chlorpromazine specifically prevents the wheel-induced feeding suppression in rats. AB - In rats, limited daytime wheel access suppresses feeding over the subsequent night [Lattanzio SB, Eikelboom R. Wheel access duration in rats: I. effects on feeding and running. Behav Neurosci 2003; 117:496-504.]. This phenomenon is known as the wheel-induced feeding suppression (WIFS). The classic antipsychotic, chlorpromazine, can minimize the severity of the related activity anorexia procedure, but is thought to act through a suppression of running [Routtenberg A. "Self-starvation" of rats living in activity wheels: adaptation effects. J Comp Physiol Psychol 1968; 66:234-8.]. We tested the effects of chlorpromazine (2mg/kg IP) on the acute WIFS in 40 adult male rats by administering the drug before or after 3h of daytime wheel access and measuring food consumption over the subsequent 24h. Control groups received saline injections or were exposed to locked wheels. While chlorpromazine did not attenuate feeding or change wheel running alone, it blocked their interaction, the acute WIFS. This procedure might be useful in screening drugs for anorexia nervosa where exercise is often elevated and feeding is suppressed. PMID- 19549533 TI - Anti-oxidative and anti-inflammatory vasoprotective effects of caloric restriction in aging: role of circulating factors and SIRT1. AB - Endothelial dysfunction, oxidative stress and inflammation are associated with vascular aging and promote the development of cardiovascular disease. Caloric restriction (CR) mitigates conditions associated with aging, but its effects on vascular dysfunction during aging remain poorly defined. To determine whether CR exerts vasoprotective effects in aging, aortas of ad libitum (AL) fed young and aged and CR-aged F344 rats were compared. Aging in AL-rats was associated with impaired acetylcholine-induced relaxation, vascular oxidative stress and increased NF-kappaB activity. Lifelong CR significantly improved endothelial function, attenuated vascular ROS production, inhibited NF-kappaB activity and down-regulated inflammatory genes. To elucidate the role of circulating factors in mediation of the vasoprotective effects of CR, we determined whether sera obtained from CR animals can confer anti-oxidant and anti-inflammatory effects in cultured coronary arterial endothelial cells (CAECs), mimicking the effects of CR. In CAECs cultured in the presence of AL serum TNFalpha elicited oxidative stress, NF-kappaB activation and inflammatory gene expression. By contrast, treatment of CAECs with CR serum attenuated TNFalpha-induced ROS generation and prevented NF-kappaB activation and induction of inflammatory genes. siRNA knockdown of SIRT1 mitigated the anti-oxidant and anti-inflammatory effects of CR serum. CR exerts anti-oxidant and anti-inflammatory vascular effects, which are likely mediated by circulating factors, in part, via a SIRT1-dependent pathway. PMID- 19549536 TI - Performance-enhancing and thermoregulatory effects of intracerebroventricular dopamine in running rats. AB - To assess the role of central dopamine on metabolic rate, heat balance and running performance, 2.0 microL of 5 x 10(-3)M dopamine solution (DA) or 0.15M NaCl (SAL) was intracerebroventricularly injected in Wistar rats 1 min before running on a motor-driven treadmill, according to a graded exercise protocol, until fatigue. Oxygen consumption (VO(2)) and body temperature (T(b)) were recorded at rest, during exercise, and after 30 min of recovery. DA induced a marked increase in workload (approximately 45%, p<0.05). At fatigue point, DA injected rats attained approximately 29% higher maximum oxygen consumption (VO(2max)) and approximately 0.75 degrees C higher T(b) than SAL-injected rats. Despite the higher VO(2max) and T(b) attained during exercise, DA-treated rats reached VO(2) basal values within the same recovery period and dissipated heat approximately 33% faster than SAL-treated rats (p<0.05). The mechanical efficiency loss rate was approximately 40% lower in DA than in SAL-treated rats (p<0.05), however, the heat storage was approximately 35% higher in the DA group (p<0.05). Our results demonstrate that increased DA availability in the brain has a performance-enhancing effect, which is mediated by improvements in the tolerance to heat storage and increases in the metabolic rate induced by graded exercise. These data provide further evidence that central activation of dopaminergic pathways plays an important role in exercise performance. PMID- 19549537 TI - Response surface optimization of polysaccharides extraction from Liriope roots and its modulatory effect on Sjogren syndrome. AB - The response surface methodology (RSM) was employed to study the extraction of liriope polysaccharides. The maximum extraction rate (64.1%) of liriope polysaccharides obtained by using the above optimised concentrations of the variables was extraction time (X(1)) at 185min, ratio of liquid to solid (X(2)) at 11, and extraction temperature (X(3)) at 94 degrees C. Forty male Wistar rats were used in the present experiment. After 2 weeks of acclimatization, animals were divided into 5 equal groups: control, SS model and 3 polysaccharides treatment groups. Rats were orally administered their respective doses every day for 6 weeks. Liriope polysaccharides significantly increased the amount of salivary secretion, and the relative weight of spleen, thymus and submandibular glands. Therefore, the present results revealed that liriope polysaccharides exert a protective effect against tissue damage in rats with sjogren syndrome (SS). PMID- 19549538 TI - Characterization and anti-tumor activities of sulfated polysaccharide SRBPS2a obtained from defatted rice bran. AB - A novel chemically sulfated polysaccharide SRBPS2a with potent anti-tumor activity was derived from defatted rice bran by chlorosulfonic acid-pyridine (CSA Pyr) method. The average molecular weight of SRBPS2a was 3.5 x 10(5) Da and the degree of sulfation (DS) was 1.29. The Fourier-transform infrared spectra (FT-IR) and 13C NMR spectroscopy analysis revealed that SRBPS2a was mainly consist of beta-(1-->3)-D-galactopyranosyl residues, the sulfate substitution site was on C 2 and C-4 while the side chains were cut off during the sulfated reaction. Furthermore, SRBPS2a exhibited evident growth inhibition on mouse mammary tumor EMT-6 cells both in vitro and in vivo. PMID- 19549539 TI - Novel chitin/nanosilica composite scaffolds for bone tissue engineering applications. AB - Biopolymers like chitin are widely investigated as scaffolds in bone tissue engineering. Its properties like biocompatibility, biodegradability, non toxicity, wound healing ability, antibacterial activity, hemostatic property, etc., are widely known. However, these materials are not much bioactive. Addition of material like silica can improve the bioactivity and biocompatibility of chitin. In this work, chitin composite scaffolds containing nanosilica were prepared using chitin hydrogel and their bioactivity, swelling ability and cytotoxicity was analyzed in vitro. These scaffolds were found to be bioactive in simulated body fluid (SBF) and biocompatible when tested with MG 63 cell line. These results suggest that chitin/nanosilica composite scaffolds can be useful for bone tissue engineering applications. PMID- 19549540 TI - Glucose-sensitivity of liposomes incorporating conjugates of glucose oxidase and poly(N-isopropylacrylamide-co-methacrylic acid-co-octadecylacrylate). AB - Liposomes, which release their contents in response to the concentration of glucose, were prepared by modifying the liposomal surface with the conjugate of poly(N-isopropylacrylamide-co-methacrylic acid-co-octadecylacrylate) (P(NIPAM-co MAA-co-ODA)) and glucose oxidase (GOD). The maximum enzymatic activity of copolymer conjugated GOD (Polym-GOD) was observed around pH 5.0 and the value was about 40% of that of native GOD. Nine lysine residues per GOD molecule, on average, were found to be covalently attached to the copolymers. Egg phosphatidylcholine liposomes bearing Polym-GOD released their contents in response to the concentration of glucose and the sensitivity was higher than dipalmitoylphosphatidylcholine liposomes. PMID- 19549541 TI - Diversity in amylopectin structure, thermal and pasting properties of starches from wheat varieties/lines. AB - Structural, thermal and pasting diversity of starches from Indian and exotic lines of wheat was studied. Majority of the starches showed amylose content ranging between 22% and 28%. Endotherm temperatures (T(o), T(p) and T(c)) of the starches showed a range between 56-57, 60 -61 and 65.5-66.5 degrees C, respectively. Exotherms with T(p) between 87.0 and 88.2 degrees C were observed during cooling of heated starches, indicating the presence of amylose-lipid complexes. Exotherm temperatures were negatively correlated to swelling power. Amylopectin unit chains with different degree of polymerization (DP) were observed to be associated with pasting temperature, setback and thermal (endothermic T(o), T(p), and T(c)) parameters. Amylopectin unit chains of DP 13 24 showed positive relationship with endothermic T(o), T(p) and T(c). Pasting temperature showed positive correlation with short chains (DP 6-12) while negative correlation with medium chain (DP 13-24) amylopectins. Setback was positively correlated to DP 16-18 and negatively to DSC amylose-lipid parameters. PMID- 19549542 TI - Structural elucidation, modification and characterization of seed gum from Cassia javahikai seeds: A non-traditional source of industrial gums. AB - A water-soluble seed gum was isolated from seed endosperm of Cassia javahikai. The acid-catalyzed fragmentation, methylation, selective enzymatic degradation and periodate oxidation suggested a heteropolymeric structure for the polysaccharide. The polysaccharide was shown to have a linear chain of beta(1- >4) linked d-mannopyranosyls units with side chains of alpha(1-->6) d galactopyranosyl units. Grafting of polyacrylamide onto the gum was performed using K(2)S(2)O(8)/ascorbic acid redox system in presence of Ag(+) as catalyst at 35+/-2 degrees C. The viscosity of the gum solution increased on grafting and the grafted gum was observed to resist biodegradation for more than 256h. Thermogravimetric analysis revealed that grafted gum was more thermally stable than native gum. PMID- 19549543 TI - Sulfonation and anticoagulant activity of botryosphaeran from Botryosphaeria rhodina MAMB-05 grown on fructose. AB - Botryosphaeran (EPS(FRU)), an exopolysaccharide of the beta-(1-->3,1-->6)-d glucan type with 31% branching at C-6, is produced by the fungus Botryosphaeria rhodina MAMB-05 when grown on fructose as carbon source. Botryosphaeran was derivatized by sulfonation to induce anticoagulant activity. The effectiveness of the sulfonation reaction by chlorosulfonic acid in pyridine was monitored by the degree of substitution and FT-IR analysis of the sulfonated EPS(FRU) (once sulfonated, EPS(FRUSULF); and re-sulfonated, EPS(FRURESULF)). Activated partial thromboplastin time (APTT) and thrombin time (TT) tests of EPS(FRURESULF) indicated significant in vitro anticoagulant activity that was dose-dependent. EPS(FRU) did not inhibit any of the coagulation tests. PMID- 19549544 TI - Different subregions of the medial preoptic area are separately involved in the regulation of copulation and sexual incentive motivation in male rats: a behavioral and morphological study. AB - The purpose of this study was to investigate whether sexual incentive motivation and copulatory performance are regulated by different subregions of the medial preoptic area (MPOA). Sexual incentive motivation was measured by means of a partner preference test. Both copulatory behavior and sexual incentive motivation were tested in male rats treated with 50mg/kg of either EGb 761 or a vehicle (distilled water) by gavage for 14 days. Administration of EGb 761 increased the number of intromissions, but had no effect on the number of mounts, mount latency, intromission latency, ejaculation latency, or post-ejaculatory interval. In the partner preference test, the total duration of visits to estrous female rats in both of the groups was significantly different from the total duration of visits to sexually active males. EGb 761 treatment increased the number of ejaculations compared both to vehicle-treated controls on day 14 and the same group on day 0. In comparison with the controls, the EGb 761-treated group showed a significant increase in the number of tyrosine hydroxylase-expressing cells in the dorsal, but not the ventral, subregion of the MPOA, and significantly high dopamine levels in the MPOA. These results indicate that EGb 761 does not affect sexual incentive motivation, but facilitates copulatory performance in male rats, suggesting that the mechanisms responsible for sexual incentive motivation and copulatory performance may be associated with differential functions of MPOA subregions. PMID- 19549545 TI - Electro-acupuncture stimulation improves motor disorders in Parkinsonian rats. AB - Electro-acupuncture (EA) is believed to be effective for alleviating motor symptoms in patients with Parkinson's disease. In a rat hemiparkinsonian model induced by unilateral transection of the medial forebrain bundle (MFB), the effects of EA stimulation were investigated. EA stimulation at a high frequency (100 Hz) significantly reduced apomorphine-induced rotational behavior. Tyrosine hydroxylase immunohistochemical staining revealed that EA at 100 Hz protected axotomized dopaminergic neurons from degeneration in the substantia nigra (SN). Moreover, high frequency EA reversed the axotomy-induced decrease in substance P content and increase in glutamate decarboxylase-67 (GAD 67) mRNA level in the midbrain; however, it did not affect the axotomy-induced increase in enkephalin content in the globus pallidus. These results suggest that the effects of high frequency EA on motor symptoms of Parkinsonian rats may involve restoration of the homeostasis of dopaminergic transmission in the basal ganglia circuit. PMID- 19549546 TI - Pharmacological effects of ethanol on ingestive behavior of the preweanling rat. AB - The present study was designed to test the hypothesis that sensitivity of ingestive behavior of infant rat to the pharmacological effects of ethanol changes between postnatal (P) days 9 and 12. The intake of 0.1% saccharin and water, general motor activity, and myoclonic twitching activity were assessed following administration of three doses of ethanol (0, 0.25, and 0.5 g/kg) while fluids were free available to the animals. The 0.5 g/kg dose of ethanol attenuated saccharin intake in P9 pups and enhanced saccharin intake in P12 rats. On P12 some sex-related differences emerged at 0.5 g/kg of ethanol, with saccharin intake being higher in females than in their male counterparts. Taste reactivity probe revealed that 0.5 g/kg of ethanol increased taste responsiveness to saccharin on P12 but only to infusions presented at a high rate. The results of the present study indicate that ontogenetic changes in sensitivity to the effects of ethanol on ingestive behavior occur during the second postnatal week, with P9 animals being more sensitive to the inhibitory (sedative) effects on saccharin intake and P12 rats being more sensitive to the stimulatory effects of ethanol. We suggest that acute ethanol enhanced saccharin intake via sensitization of oral response to appetitive taste stimulation. PMID- 19549548 TI - Imbalance in oxidant/antioxidant system in different brain regions of rat after the infection of Japanese encephalitis virus. AB - The imbalance in redox equilibrium is associated with several viral diseases however its role in Japanese encephalitis (JE) has not been reported. In the present study, we report the status of oxidant/antioxidant system in different brain regions in rat model of JE. Twelve days old Wistar strain rats were inoculated intracerebrally with a dose of 3x10(6)pfu of JE virus (JEV). The activity of manganese superoxide dismutase (Mn-SOD), catalase (CAT), glutathione peroxidase (GPx) and the levels of reduced glutathione (GSH) and malonaldialdehyde (MDA) were estimated in corpus striatum, frontal cortex, thalamus and midbrain on 0, 10 and 20 days post-inoculation (dpi). A significant increase in MDA levels in striatum (p<0.01), cortex (p<0.01), thalamus (p<0.01) and midbrain (p<0.01) was observed in JEV infected rats on 10 and 20dpi compared to controls. The activity of CAT, GPx and the levels of GSH were significantly decreased in all the brain regions studied on 10 and 20dpi compared to controls. However, the activity of Mn-SOD in striatum (p<0.01), cortex (p<0.05), thalamus (p<0.01) and midbrain (p<0.01) were significantly increased on 10 and 20dpi in JEV infected rats compared to controls. The activity of Mn-SOD and MDA levels were significantly increased whereas the activity of CAT, GPx and GSH levels were significantly decreased in all the brain regions studied as the disease progressed from 0 to 20dpi. The maximum alteration in oxidant/antioxidant balance was observed in thalamus and midbrain. The results of the present study demonstrate that antioxidant defense mechanism is impaired after the infection of JE virus suggesting its critical role in cellular injury in brain regions. The findings could be beneficial to understand the role of oxidative stress in the pathogenesis of JE and therapeutic interventions. PMID- 19549549 TI - Neurosteroids modulate compulsive and persistent behavior in rodents: implications for obsessive-compulsive disorder. AB - Neurosteroids are reported to modulate GABAergic and glutamatergic pathways that then influence serotonin and dopamine, the neurotransmitters implicated in pathophysiology of obsessive-compulsive disorder (OCD). Fluoxetine, a selective serotonin reuptake inhibitor clinically used in OCD is reported to increase the levels of neurosteroids like allopregnanolone, whereas OCD patients exhibit higher plasma levels of dehydroepiandrosterone 3-sulphate (DHEAS), a neuroactive steroid having opposite effects to that of allopregnanolone. Hence, it was contemplated that neurosteroids may influence obsessive-compulsive behavior. To test this possibility we studied the influence of various neurosteroids on two behavioral models of OCD, namely marble-burying behavior in mice and 8-OH-DPAT induced disruption of spontaneous alternation behavior (SAB) in rats. The results revealed that allopregnanolone (1 microg/mouse, i.c.v) and progesterone (20mg/kg, s.c.) reduced the marble-burying behavior in mice, whereas dehydroisoandrosterone 3-sulphate (DHAS) (5mg/kg, i.p.) exacerbated the same. The effects of allopregnanolone were comparable to that of fluoxetine (10mg/kg, i.p.). In view of the report that restraint stress increases the levels of allopregnanolone and isolation stress decreases the same, we studied the effect of these stressors on marble-burying behavior; wherein it was found to be less in restraint stress exposed mice, and higher in socially isolated mice. Restrain stress-induced attenuation of marble-burying behavior was blocked by finasteride, a neurosteroid biosynthesis blocker. In rat model of SAB disruption, acute and chronic treatment with allopregnanolone (1 microg/mouse, i.c.v.) reduced 8-OH-DPAT-induced persistent behavior, whereas treatment with DHAS (5mg/kg, i.p.) had an opposite effect. In conclusion, the studies indicate that neurosteroids can modulate obsessive-compulsive behavior in a bidirectional manner, and could serve as an effective target in the management of OCD. PMID- 19549547 TI - The changing role of the medial preoptic area in the regulation of maternal behavior across the postpartum period: facilitation followed by inhibition. AB - Maternal behavior in rats undergoes considerable plasticity in parallel to the developmental stage of the pups, resulting in distinct patterns of maternal behavior and care at different postpartum time points. The medial preoptic area (mPOA) of the hypothalamus is one critical neural substrate underlying the onset and early expression of maternal behavior in rats but little is known about its specific functional role in the evolving expression of maternal behavior across the postpartum period. The present study uses a reversible local neural inactivation method to examine the role of the mPOA in the regulation of maternal behavior throughout the postpartum period, particularly extending into the late postpartum, a little examined period. This approach avoids the compensatory plasticity in CNS that occurs after permanent lesions, and allows the repeated testing of same individuals. Early (PPD7-8) and late (PPD13-14) postpartum maternal behavior was evaluated in female rats following infusions of bupivacaine or vehicle into the mPOA or into control areas. As expected, mPOA inactivation severely but transiently disrupted early postpartum maternal behavior whereas infusion of vehicle or inactivation of adjacent control sites did not. Later in the postpartum period, however, transient mPOA inactivation facilitated the expression of maternal behaviors, highly contrasting the behavioral expression levels characteristic of late postpartum. Results strongly demonstrate that the mPOA is differentially engaged throughout postpartum in orchestrating appropriate maternal responses with the developmental stage of the pups. PMID- 19549550 TI - Environmental enrichment improves cognitive deficits in Spontaneously Hypertensive Rats (SHR): relevance for Attention Deficit/Hyperactivity Disorder (ADHD). AB - The interaction between genes and environment seems to be relevant for the development of Attention Deficit/Hyperactivity Disorder (ADHD), one of the most prevalent childhood psychiatric diseases. The occurrence of ADHD is typically associated with poor academic performance, probably reflecting learning difficulties and/or cognitive impulsiveness. The inbred Spontaneously Hypertensive Rats (SHR) strain has often been considered as an animal model of ADHD, since they 'naturally' display the main ADHD symptomatology. Although pharmacological agents improve SHR's cognitive deficits, little is known about the involvement of environmental factors in SHR disabilities and to what extent 'protective' non-pharmacological factors may be considered as strategy for ADHD prevention. Here we investigated whether the rearing environment during neurodevelopment may counteract later cognitive deficits presented by adult SHR. Wistar (WIS) rats were also used to investigate whether the putative effects of environmental enrichment depend on a specific genetic background. The animals were reared in enriched environment (EE) or standard environment (SE) from the post-natal day 21 until 3 months of age (adulthood) and tested for cognitive and non-cognitive phenotypes. EE improved SHR's performance in open field habituation, water maze spatial reference, social and object recognition tasks, while non-cognitive traits, such as nociception and hypertension, were not affected by EE. Response of WIS rats was generally not affected by the present EE. These results show that the general low cognitive performance presented by SHR rats strongly depends on the rearing environment and they may suggest modifications of the familial environment as a putative preventive strategy to cope with ADHD. PMID- 19549551 TI - Radioactivity and heavy metal levels in hazelnut growing in the Eastern Black Sea Region of Turkey. AB - The Eastern Black Sea Region of Turkey is one of the main hazelnut producers in Turkey and in the world. Since this region was contaminated by the Chernobyl accident in 1986, a comprehensive study was planned and carried out to determine the radioactivity level in hazelnut growing region. The dose due to consumption of hazelnut by the public was estimated and it was shown that this dose imposes no threat to human health. In addition, heavy metal analysis was performed in the samples and the amount of Cr, Mn, Fe, Ni, Cu, Zn, and Pb were also detected. The results showed that the concentrations of heavy metal are below the daily intake recommended by the international organizations. PMID- 19549552 TI - Long-term effects of developmental exposure to di-n-butyl-phthalate (DBP) on rat prostate: proliferative and inflammatory disorders and a possible role of androgens. AB - In the present study we evaluated the toxic effects on the male adult rat prostate of DBP exposure during fetal and lactational periods, because although many studies have addressed the influence of phthalates on the male reproductive system, only a few have discussed their possible effects on prostate development. Pregnant females were distributed into two experimental groups: Control (C) and Treated (T). The females of the T group received DBP (100mg/kg, by gavage) from gestation day 12 to postnatal day 21, while C rats received the vehicle (corn oil). In adulthood (90 days old), the animals were euthanized. The serum and testicular testosterone levels were measured. Ventral prostate was removed and weighed. Distal segment fragments of the ventral prostate were fixed and processed for histochemistry and immunohistochemistry to detect androgen receptor (AR) and Ki67 antigens. Protein extraction from ventral prostate fragments was performed for AR immunoblotting and Gelatin zymography for MMP-2 and MMP-9 (MMP, metalloproteinase). Stereological and histopathological analyses were also performed. Serum and testicular testosterone levels and prostate weight were comparable between groups. In the T group the relative proportions (%) of epithelial (C=32.86; T=42.04*) and stromal (C=21.61; T=27.88*) compartments were increased, while the luminal compartment was decreased (C=45.54; T=30.08*), *p<0.05. InT, disseminated inflammatory infiltrate in the stroma, associated or not with epithelial dysplasia and PIN (Prostatic Intraepithelial Neoplasia), was observed. Increases in AR expression, proliferation index and metalloproteinase 9 (MMP-9) activity were noted in T animals. In some T animals, collagen fibrils accumulated adjacent to the epithelium. As far as we are aware, this is the first report in the literature showing that phthalates could play a role in proliferative and inflammatory disorders of the rat prostate. PMID- 19549553 TI - The mycotoxin deoxynivalenol inhibits the cell surface expression of activation markers in human macrophages. AB - Deoxynivalenol (DON) is the most prevalent trichothecene mycotoxin in crops in Europe and North America. It exhibits several toxic effects including impaired growth and immune dysregulation. Macrophages play pivotal role in the host defense; upon activation, they express several specific cell surface receptors that are important in adhesion and cell signaling. Several studies have demonstrated that DON can affect macrophages, however, very few data are available concerning the effect of DON on human macrophages, and the effect on macrophage cell surface receptors is unknown. In the present study, human blood monocytes, differentiated in vitro into macrophages, were activated with IFN gamma, in the presence or absence of low concentrations of DON. The expression of CD11c, CD13, CD14, CD18, CD33, CD35, CD54, CD119 and HLA-DP/DQ/DR was analyzed by flow cytometry. As expected, macrophage activation by IFN-gamma upregulated the expression of CD54, CD14, CD119 and HLA-DP/DQ/DR. Incubation with DON decrease the cell surface expression of these activation markers in a dose-dependent manner. When cells were treated with 5muM DON, the mean fluorescence intensity measured for the expression of these receptors was the same as that observed in non-activated macrophages. This inhibitory effect of DON was only observed when the mycotoxin was applied before the activation signal. Taken together, our results suggest that low concentration of DON alter macrophage activation as measured by the expression of cell surface markers. This may have implications for human health when consuming DON contaminated feed. PMID- 19549555 TI - Involvement of prostaglandin F2alpha in the adverse effect of PCB 77 on the force of contractions of bovine myometrium. AB - Polychlorinated biphenyls (PCBs) stimulate in vitro both the force of myometrial contractions and endometrial secretion of PGF2alpha in cattle. Therefore, the goal of this study was to investigate the participation of PGF2alpha in the effect of PCBs on uterine contractility. For this aim, the myometrial strips were incubated (48h) with PCB 77 at the dose of 1, 10 and 100ng/ml (i.e., 0.0034, 0.034 and 0.34nmol/ml) separately or jointly with indomethacin (INDO, 10(-4)M), which blocks the PGF2alpha synthesis. Next, the force of myometrial strips contractions was measured. Further, the influence of PCB 77 (0.1, 1 and 10ng/ml) on the PGF2alpha secretion from myometrial cells after 6, 24, and 48h and PCB 77 (1 and 10ng/ml) on the mRNA expression of cyclooxygenase 2 (COX-2) and PGF2alpha synthase (PGFS) in myometrial cells after 6 and 24h, was investigated. The increase (P<0.05-0.001) of the contractions force of myometrial strips evoked by each dose of PCB 77, was markedly reduced (P<0.05-001) by INDO. There was an increase (P<0.05-0.001) of both PGF2alpha secretion after all studied periods of cell incubation and mRNA expression for COX-2 and PGFS after 6h treatment of myometrial cells with PCB 77. It can be concluded that myometrial synthesis of PGF2alpha and its further secretion is a part of the mechanism by means of which PCB 77 may affect the force of myometrial contractions in cattle. PMID- 19549556 TI - Coupling of mesoscopic brain oscillations: recent advances in analytical and theoretical perspectives. AB - Oscillatory brain activities have been traditionally studied in the context of how oscillations at a single frequency recorded from a single area could reveal functional insights. Recent advances in methodology used in signal analysis have revealed that cross-frequency coupling, within or between functional related areas, is more informative in determining the possible roles played by brain oscillations. In this review, we begin by describing the cellular basis of oscillatory field potentials and its theorized as well as demonstrated role in brain function. The recent development of mathematical tools that allow the investigation of cross-frequency and cross-area oscillation coupling will be presented and discussed in the context of recent advances in oscillation research based on animal data. Particularly, some pitfalls and caveats of methods currently available are discussed. Data generated from the application of examined techniques are integrated back into the theoretical framework regarding the functional role of brain oscillations. We suggest that the coupling of oscillatory activities at different frequencies between brain regions is crucial for understanding the brain from a functional ensemble perspective. Effort should be directed to elucidate how cross-frequency and area coupling are modulated and controlled. To achieve this, only the correct application of analytical tools may shed light on the intricacies of information representation, generation, binding, encoding, storage and retrieval in the brain. PMID- 19549554 TI - Liquid chromatography electrospray ionization tandem mass spectrometry analysis method for simultaneous detection of trichloroacetic acid, dichloroacetic acid, S (1,2-dichlorovinyl)glutathione and S-(1,2-dichlorovinyl)-L-cysteine. AB - Trichloroethylene (TCE, CAS 79-01-6) is a widely used industrial chemical, and a common environmental pollutant. TCE is a well-known carcinogen in rodents and is classified as "probably carcinogenic to humans". Several analytical methods have been proposed for detection of TCE metabolites in biological media utilizing derivatization-free techniques; however, none of them is suitable for simultaneous detection of both oxidative metabolites and glutathione conjugates of TCE in small volume biological samples. Here, we report a new combination of methods for assessment of major TCE metabolites: dichloroacetic acid (DCA), trichloroacetic acid (TCA), S-(1,2-dichlorovinyl)-L-cysteine (DCVC), and S-(1,2 dichlorovinyl) glutathione (DCVG). First, DCA and TCA were extracted with ether. Second, the remaining aqueous fraction underwent solid phase extraction for DCVC and DCVG. Then, DCA and TCA were measured by hydrophilic interaction liquid chromatography ion exchange negative electrospray ionization tandem mass spectrometry, while DCVC and DCVG were measured by reverse phase positive electrospray ionization tandem mass spectrometry. This method was applied successfully to measure all 4 TCE metabolites in as little as 50 microl of serum from mice orally exposed to TCE (2100 mg/kg, 2h). Serum concentrations (mean+/ standard deviation) of the TCE metabolites obtained with this method are comparable or equivalent to those previously reported in the literature: DCA, 0.122+/-0.014 nmol/ml (limit of detection: 0.01 nmol/ml); TCA, 256+/-30 nmol/ml (0.4 nmol/ml); DCVG, 0.037+/-0.015 nmol/ml (0.001 nmol/ml); DCVC, 0.0024+/-0.0009 nmol/ml (0.001 nmol/ml). This method opens new opportunities to increase throughput and decrease number of animals required for mechanistic studies on TCE in rodents. PMID- 19549557 TI - Voltage-gated delayed rectifier K v 1-subunits may serve as distinctive markers for enteroglial cells with different phenotypes in the murine ileum. AB - Due to entangled results concerning K(v)1 subunit distribution in the gastrointestinal wall, we aimed to unravel the expression of the delayed rectifier potassium subunits K(v)1.1 and K(v)1.2 in the murine ileum. Presence and distribution of both subunits were determined in cryosections and whole-mount preparations of the ileum of three different murine strains by indirect immunofluorescence, and analysed by conventional fluorescence and confocal microscopy. Distribution of both subunits was similar in the ileum of the three strains. K(v)1.1 immunoreactivity (IR) was found in some S100-expressing enteroglial cells (EGC) located at the periphery of myenteric ganglia, in S100 positive EGC along interganglionic, intramuscular and vascular nerve fibres, and in S100-positive EGC of the submucous plexus. K(v)1.1 IR was also observed in some GFAP-expressing EGC at the periphery of myenteric ganglia, and in GFAP positive EGC of submucous ganglia. K(v)1.2 IR was detected in some intramuscular S100-positive EGC, in almost all submucous S100-expressing EGC, and in a few GFAP expressing EGC. K(v)1.2 IR was also expressed in a majority of enteric neurons. Coding of these neurons showed that all cholinergic and most nitrergic neurons express K(v)1.2. In conclusion, the results showed that K(v)1.1 and K(v)1.2 were predominantly expressed in distinct EGC phenotypes. K(v)1.2 was also observed in distinct neuron subpopulations. Our results support the active role of EGC with distinct phenotypes in intestinal functions, which is relevant in view of their modulating role on intestinal barrier and inflammatory responses. PMID- 19549558 TI - Adipose stromal cells-secreted neuroprotective media against neuronal apoptosis. AB - Transplantation of pluripotent adipose stem/stromal cells (ASC) alleviates tissue damage and improves functional deficits in both stroke and cardiovascular disease animal models. Recent studies indicate that the primary mechanism of ASC-induced repair may not be directly related to tissue regeneration through differentiation, but rather through paracrine mechanisms provided by secreted pro survival and repair-inducing trophic factors. In this study, we have found that ASC-conditioned medium (ASC-CM) potently protected cerebellar granule neurons (CGN) from apoptosis induced by serum and potassium deprivation. Neural cell protection was mostly attributable to activated caspase-3 and Akt-mediated neuroprotective pathway signaling. Specific neutralization of neurotrophic factor activity demonstrated that serum and potassium deprivation-induced Akt-mediated neuroprotection and caspase-3-dependent apoptosis were mainly modulated by IGF-1. These data suggest that of the many neuroprotective factors secreted by ASC, IGF 1 is the major factor that mediates protection against serum and potassium deprivation-induced CGN apoptosis. This study establishes a mechanistic basis supporting the therapeutic application of ASC for neurological disorders, specifically through paracrine support provided by trophic factor secretion. PMID- 19549559 TI - SVM-based computer-aided diagnosis of the Alzheimer's disease using t-test NMSE feature selection with feature correlation weighting. AB - This letter shows a computer-aided diagnosis (CAD) technique for the early detection of the Alzheimer's disease (AD) based on single photon emission computed tomography (SPECT) image feature selection and a statistical learning theory classifier. The challenge of the curse of dimensionality is addressed by reducing the large dimensionality of the input data and defining normalized mean squared error features over regions of interest (ROI) that are selected by a t test feature selection with feature correlation weighting. Thus, normalized mean square error (NMSE) features of cubic blocks located in the temporo-parietal brain region yields peak accuracy values of 98.3% for almost linear kernel support vector machine (SVM) defined over the 20 most discriminative features extracted. This new method outperformed recent developed methods for early AD diagnosis. PMID- 19549560 TI - Observing shadow motions: resonant activity within the observer's motor system? AB - Several studies have demonstrated that the human motor cortex is activated by the mere observation of actions performed by others. In the present study, we explored whether the perception of 'impoverished motion stimuli', such as shadow animations, is sufficient to activate motor areas. To do so, transcranial magnetic stimulation (TMS) was applied over the hand area of the primary motor cortex (M1) while subjects observed shadow animations depicting finger motions. Data showed that resonant motor responses in M1 were only found when a biological effector was recognized from the observed shadow animation. Interestingly, M1 responses were similar for observing shadow or real motions. Therefore, the loss of 'pictorial' movement features in a shadow animation appeared to have no effect on motor resonance in M1. In summary, these findings suggest that the 'recognition' of biological motion from sparse visual input is both necessary and sufficient to recruit motor areas. This supports the hypothesis that the motor system is involved in recognizing the actions performed by others. PMID- 19549561 TI - Neuroprotective effects of group II metabotropic glutamate receptor agonist DCG IV on hippocampal neurons in transient forebrain ischemia. AB - Activation of group II metabotropic glutamate receptor (mGluR) inhibits the excessive release of glutamate that may be crucial in the pathogenesis of cerebral ischemia. This study investigated the protective effects of the group II mGluR agonist (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG-IV), against cerebral ischemia by examining extracellular glutamate concentration ([Glu]e) and neuronal damage in a rat model of transient forebrain ischemia. Cerebral ischemia was induced by 5 min of bilateral carotid artery occlusion and hypotension. DCG-IV (10, 100, or 250 pmol) was administered into the lateral ventricle four times every 12 h from 36 h before the start of ischemia, or administered intraperitoneally (40 micromol/kg) 24 h before ischemia, and the effect of the group II mGluR antagonist (LY341495) was also examined. [Glu]e in the CA1 subfield was measured by microdialysis during the peri-ischemic period, and the survival rate of CA1 neurons was evaluated 5 days after ischemia. [Glu]e increased significantly after cerebral ischemia and reached the maximum at 1 min after reperfusion, then gradually decreased and returned to the preischemic level in the vehicle group. The intraventricular injection of DCG-IV (250 pmol) significantly attenuated the [Glu]e increase and significantly increased the survival rate of CA1 neurons. Co-injection of LY341495 reversed the protective effects of DCG-IV. These results suggest that pretreatment with DCG-IV has neuroprotective effects against ischemic neuronal injuries through the inhibition of the glutamate release via the activation of group II mGluR. PMID- 19549562 TI - Effect of Astragali-Cordyceps Mixtura on TGF-beta/Smad signal pathway in the lung of asthma airway remodeling. AB - AIM OF THE STUDY: We try to find out the influence of traditional Chinese Medicine Astragali-Cordyceps Mixtura (ACM) on TGF-beta/Smad signal pathway in the lung of asthma airway remodeling. MATERIALS AND METHODS: Mice were sensitized and challenged by OVA to establish a model of asthma. To assess the effects of ACM on the mice, animals of the ACM groups were treated with ACM. Data were achieved by using techniques as follow: counting cell number of BALF, assaying the amount of collagen deposition by Masson's staining, performing RT-PCR and immunohistochemistry for mRNA and protein expression of TGF-beta1, Smad3 and Smad7. RESULTS: The depositions of collagen in airway wall greatly increased at the model group compared with that of the normal group. In contrast, these decreased at the ACM groups. As compared with the control group, TGF-beta1 expression also decreased at both mRNA and protein level at the ACM-M group versus increased both at the model group. Whereas, Smad7 significantly decreased only at the model group and partly restored at the ACM-M group. CONCLUSIONS: ACM greatly improves the symptoms of asthma airway remodeling by inhibiting the expression of TGF-beta1 and upregulating the amount of Smad7. PMID- 19549563 TI - A traditional Chinese medicine formula extracts stimulate proliferation and inhibit mineralization of human mesenchymal stem cells in vitro. AB - AIM OF THE STUDY: To investigate the effects of a traditional Chinese medicine (TCM) formula extract, named as ZD-I, on the proliferation and osteogenic differentiation of human mesenchymal stem cells (hMSCs) in vitro. MATERIALS AND METHODS: When hMSCs cultivated in the basal medium with ZD-I, cell viability was assessed by MTT assay and cellular proliferation was assessed by SYBR green I assay. The effects of ZD-I on osteogenic differentiation of hMSCs were assessed by alkaline phosphatase (ALP) activity, mineralization assay and real-time RT PCR. RESULTS: ZD-I (0.78-100 microg/ml) was non-cytotoxic. The 50% inhibitory concentration (IC50) of hMSCs was 200 microg/ml. ZD-I (0.78-50 microg/ml) stimulated the proliferation of hMSCs. ZD-I did not change ALP activity of hMSCs cultivated in osteogenic medium in the early stage (4 and 7 days), but ZD-I inhibited the mineralization of hMSCs through down-regulation of several osteogenic markers (e.g. osteocalcin, bone morphogenetic protein 2 and osteopontin) in the late stage. CONCLUSIONS: ZD-I stimulate cellular proliferation and decrease the bone mineral deposition of hMSCs. These results suggest ZD-I may play an important therapeutic role in osteoarthritic patients by improving proliferative capacity of hMSCs and inhibiting the mineralization of hMSCs. PMID- 19549564 TI - Dual effects of lipophilic extract of Salvia miltiorrhiza (Danshen) on catecholamine secretion in cultured bovine adrenal medullary cells. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Salvia miltiorrhiza (Danshen) is a well known traditional Chinese herb, which has been used widely in China for the treatment of cardiovascular diseases in clinic. AIM OF THIS STUDY: The aim of the present study is to clarify the effects of lipophilic extract of Salvia miltiorrhiza (LESM) on catecholamine (CA) secretion, a traditional Chinese medicine used widely for the treatment of cardiovascular diseases in China. MATERIALS AND METHODS: LESM was evaluated for its effects on CA secretion using HPLC-ECD method. The effects of LESM on 22Na+ influx and intracellular calcium ([Ca2+]i) were also investigated. RESULTS: Our results showed that LEMS directly stimulated basal CA secretion in an extracellular Ca2+-dependent manner. And the stimulation was not affected by combination of hexamethonium (Hex),an inhibitor of nAChR. LESM also directly elevated [Ca2+]i. In addition, using selective blockers of voltage-dependent Ca2+ channels, such as nitrendipine (for L-type), omega agatoxin-IVA (for P-type) and -conotoxin-GVIA (for N-type), it was found that nitrendipine suppressed the elevation of [Ca2+]i induced by LESM, but not omega agatoxin-IVA or omega-conotoxin-GVIA. Compared with acetylcholine (ACh) only, however, combination of LESM with ACh inhibited the raise of CA secretion, 22Na+ influx and [Ca2+]i in a concentration-dependent manner. Furthermore, LESM also inhibited CA secretion induced by veratridine (Ver), and 56 mM K+ at concentrations similar to those for [Ca2+]i rise. One of the lipophilic active compounds, cryptotanshione (Cryp), also had the same effects on CA secretion with LESM. CONCLUSIONS: All these findings suggest that LESM exerts dual effects on CA secretion in cultured bovine adrenal medullary cells. LESM exerts antagonistic effects on nAChR, voltage-dependent Na+ and Ca2+ channels, whereas it is an agonist of L-type Ca2+ channel when it used alone. PMID- 19549565 TI - Antibacterial activity of Helleborus bocconei Ten. subsp. siculus root extracts. AB - AIM OF THE STUDY: Roots of Helleborus bocconei Ten. subsp. siculus (Schiffner) Merxm. & Podl. are widely used in veterinary folk medicine in Sicily (Italy) to diagnose and cure lower respiratory tract infections in cattle. This study intended to evaluate the in vitro antibacterial activity of the methanolic root extract of Helleborus bocconei Ten. subsp. siculus, and of the bufadienolide and ecdysteroid fractions extracted from its roots, against strains belonging to species commonly associated with respiratory tract infections. MATERIALS AND METHODS: The phytochemical screening of the previously prepared plant extracts was carried out by chemical, thin-layer chromatography and spectroscopic methods. The in vitro antibacterial activity of the extracts against seven different standard bacterial strains was evaluated by broth microdilution. RESULTS: According to the present study, Helleborus bocconei Ten. subsp. siculus roots contain bufadienolides and ecdysteroids, and the extracts containing these compounds, as well as the crude methanolic root extract, show antibacterial activity against microorganisms responsible for respiratory infections. In particular, the bufadienolide extract has the highest inhibitory activity against all the tested organisms, and, as the other extracts, shows the lowest MIC values (100mug/ml) against Moraxella catarrhalis and Streptococcus pneumoniae. CONCLUSIONS: Since the roots of Helleborus bocconei Ten. subsp. siculus contain substances with antibacterial activity, the traditional use of this plant may also derive from its antibacterial properties. PMID- 19549566 TI - Superovulation in mice alters the methylation pattern of imprinted genes in the sperm of the offspring. AB - Some steps of the assisted reproduction techniques, such as superovulation, may interfere with imprinting reprogramming. In the present study, superovulation was induced in the mouse and its possible effects on the differentially methylated domains of 2 paternally (H19 and Gtl2) and 3 maternally (Peg1, Snrpn and Peg3) imprinted genes were tested in the male offspring over 2 generations. The CpGs methylation status was analyzed by pyro- and bisulfite sequencing. In liver, skeletal muscle and tail, no effect of superovulation could be observed. In the sperm, however, a significant 6% decrease in the number of methylated CpGs of H19 and significant 2.8- and 7.0-fold increases in those of Peg1 and Snrpn, respectively were observed following superovulation. The changes were still present in the H19 and Snrpn genes of the second generation offspring. This suggests that superovulation in the mother transgenerationally affects the offspring sperm methylation pattern. PMID- 19549567 TI - Maps of cardiovascular and respiratory regions of rat ventral medulla: focus on the caudal medulla. AB - The ventral medulla oblongata is critical for cardiorespiratory regulation. Here we review previous literature relating to sites within the ventral medulla that have been identified as having a 'cardiovascular' or 'respiratory' function. Together with the maps generated here, of sites from which cardiovascular and respiratory responses were evoked by glutamate microinjection, specific 'cardiovascular' regions have been defined and delineated. Commonly investigated regions, including the vasopressor rostral ventrolateral medulla (RVLM) and vasodepressor caudal ventrolateral medulla (CVLM), or areas only described by others, such as the medullary cerebral vasodilator area, are included for completeness. Emphasis is given to the caudal medulla, where three pressor regions, the caudal pressor area (CPA), the intermediate pressor area (IPA) and the medullo-cervical pressor area (MCPA), caudal to the vasodepressor CVLM were defined in the original data provided. The IPA is most responsive under pentobarbitone rather than urethane anaesthesia clearly delineating it from both the rostrally located CPA and the caudally located MCPA. The description of these multiple pressor areas appears to clarify the confusion that surrounds the identification of the 'CPA'. Also noted is a vasopressor region adjacent to the vasodepressor CVLM. Apart from the well described ventral respiratory column, a region medial to the pre-Botzinger is described, from which increases in both phrenic nerve frequency and amplitude were evoked. Limitations associated with the technique of glutamate microinjection to define functionally specific regions are discussed. Particular effort has been made to define and delineate the regions with respect to ventrally located anatomical landmarks rather than the commonly used ventral surface or dorsal landmarks such as the obex or calamus scriptorius that may vary with the brain orientation or histological processing. This should ensure that a region can easily be defined by all investigators. Study of defined regions will help expedite the identification of the role of the multiple cell groups with diverse neurotransmitter complements that exist even within each of the regions described, in coordinating the delivery of oxygenated blood to the tissues. PMID- 19549568 TI - Structure-function correlation of micro1 for micromere specification in sea urchin embryos. AB - The micromeres of sea urchin embryos have two functions: to promote the autonomous differentiation of skeletogenic cells and to induce endomesodermal tissues. Micromere specification is controlled by a double-repression gate consisting of two repressors, Pmar1 and HesC. Micro1/pmar1 encodes a transcriptional repressor with a paired-type N-terminal homeodomain and two C terminal serine-rich repeats, each of which includes a sequence similar to engrailed homology region 1, which interacts with the co-repressor Groucho. To understand the molecular mechanisms of the double-repression gate, we examined the correlation between the structure and function of micro1. Phenotypic and gene expression pattern analyses of embryos injected with mutated micro1 mRNA revealed that micro1 consists of five functional domain and motifs; namely, a DNA-binding homeodomain, a nuclear localization signal in the C-terminal flanking region of the homeodomain, and two eh1-like motifs plus a short C-terminal stretch that together mediate transcriptional repression. Our data suggest that micro1 represses target genes, including hesC, via two redundant means: its eh1-like and C-terminal motifs. The C-terminal motif requires unidentified sequences for micro1 function; a micro1 mutant with the motif but lacking the unidentified sequences failed to trigger the double-repression gate for early micromere regulatory genes, except for delta, though it did repress hesC. Our results suggest that the spatial regulation of primary mesenchyme cell specification genes, including tbr, alx1, and ets1, may be different from that of delta. PMID- 19549569 TI - Modeling drug release from hot-melt extruded mini-matrices with constant and non constant diffusivities. AB - Different types of ethylcellulose-based mini-matrices were prepared by hot-melt extrusion and thoroughly characterized in vitro. Metoprolol tartrate was used as model drug, and various amounts and types of polyethylene glycol (PEG)/polyethylene oxide (PEO) were added as release rate modifiers. Based on the experimental results, appropriate mathematical theories were identified/developed, allowing for a better understanding of the underlying drug release mechanisms. For instance, it could be shown that at high initial PEG/PEO contents and/or intermediate initial PEG/PEO contents of low molecular weight, drug diffusion with time- and position-independent diffusivities is predominant. In contrast, at low initial PEG/PEO contents and intermediate initial PEG/PEO contents of high molecular weight, the time- and position-dependent dynamic changes in the matrix porosities significantly affect the conditions for drug and PEG/PEO diffusion. These dynamic changes must be taken into account in the mathematical model. Importantly, the proposed theories are mechanistic realistic and also allow for the quantitative prediction of the effects of the device design on the resulting drug release patterns. Interestingly, these quantitative predictions could be confirmed by independent experiments. Furthermore, Raman spectroscopy allowed for the determination of the resulting drug concentration position profiles within the mini-matrices as a function of time and confirmed the theoretical predictions. PMID- 19549570 TI - The demise of subfamily Hydrocotyloideae (Apiaceae) and the re-alignment of its genera across the entire order Apiales. AB - As circumscribed by Drude, the umbellifer subfamily Hydrocotyloideae posed a major hindrance to resolving the phylogeny of order Apiales. Previous studies have suggested its polyphyly, but have not had sufficient sampling to address the issue fully. To put an end to the out-dated concept of Hydrocotyloideae, we investigated the placement of 40 of the 42 genera once placed in the subfamily, using extensive taxon sampling across the entire order. Molecular phylogenies were constructed using plastid sequences of the rpl16 intron and the trnD-trnT regions and revealed at least six hydrocotyloid lineages dispersed across both families Apiaceae and Araliaceae. The most speciose of these clades corresponds to the recently erected subfamily Azorelloideae. Another lineage includes genera grouped in Mackinlayoideae, where relationships are well resolved. Platysace appears paraphyletic with respect to Homalosciadium, and their placement is well supported as a basal lineage in Apiaceae. The type genus, Hydrocotyle, belongs to a supported clade in Araliaceae. The placements of Hermas as sister to a clade consisting of Apiaceae subfamilies Apioideae and Saniculoideae, and of Choritaenia as sister to Lichtensteinia in a clade with affinities to both Apioideae and Saniculoideae, calls into question the circumscriptions of the two subfamilies. Finally, plastid data suggest that many former hydrocotyloid genera are non-monophyletic (e.g., Azorella, Schizeilema, and Eremocharis) and are in dire need of additional phylogenetic and taxonomic studies. PMID- 19549571 TI - Curcumin prevents the oxidation and lipid modification of LDL and its inhibition of prostacyclin generation by endothelial cells in culture. AB - Low-density lipoprotein (LDL) was isolated from human plasma and oxidized by 5microM copper sulfate for 4h at 37 degrees C in the absence and presence of 1, 3, 5, 10, or 20microM of curcumin. LDL oxidized in the absence of curcumin (oxLDL) showed an increased levels of conjugated dienes, lipid peroxides (TBARS) and lysolecithin (lysoPC) and a significant loss of polyunsaturated fatty acids (PUFA). LDL oxidized with 5microM copper sulfate in the presence of curcumin caused a significant decrease of conjugated diene, lipid peroxides, lysoPC and significant increase of PUFA compared to oxLDL. These changes were dose dependent and reached a maximum at 5microM curcumin. Incubation of human endothelial cells (EC) with 200microg protein/ml of oxLDL caused a significant decrease of prostacyclin (PGI(2)) generation. LDL oxidized in presence of 5microM curcumin did not show any inhibition of PGI(2) generation compared to the control cells. These results indicate that curcumin is an effective chain-breaking antioxidant which prevents oxidation and lipid modification of LDL. The inhibition of oxLDL on PGI(2) is considered a contributing factor in the pathogenesis of thrombosis and atherosclerosis. Curcumin supplementation could be an effective strategy in preventing LDL oxidation and its impact on atherosclerosis and lesion formation. PMID- 19549573 TI - Advances and challenges in infectious diseases supportive care of patients with hematologic malignancies, hematopoietic stem cell transplantation, and severe aplastic anemia. AB - Infectious diseases are important causes of morbidity and mortality in immunocompromised patients with hematological malignancies, severe aplastic anemia (SAA), and myelodysplasia. Major advances in infectious diseases supportive care have been critical to improving the outcome of patients suffering from these life-threatening diseases. Advances in diagnosis, treatment, and prevention of life-threatening infections have reduced morbidity and mortality, improved quality of life, and enabled the use of potentially curative chemotherapy, radiation, hematopoietic stem cell transplantation (HSCT), and immunosuppressive therapy to patients battling these devastating diseases. Despite these advances, the continued development of antimicrobial resistance, emergence of new pathogens, and the evolution of host factors present evolving challenges to the successful management of infectious complications in this expanding patient population. PMID- 19549572 TI - What can mitochondrial heterogeneity tell us about mitochondrial dynamics and autophagy? AB - A growing body of evidence shows that mitochondria are heterogeneous in terms of structure and function. Increased heterogeneity has been demonstrated in a number of disease models including ischemia-reperfusion and nutrient-induced beta cell dysfunction and diabetes. Subcellular location and proximity to other organelles, as well as uneven distribution of respiratory components have been considered as the main contributors to the basal level of heterogeneity. Recent studies point to mitochondrial dynamics and autophagy as major regulators of mitochondrial heterogeneity. While mitochondrial fusion mixes the content of the mitochondrial network, fission dissects the mitochondrial network and generates depolarized segments. These depolarized mitochondria are segregated from the networking population, forming a pre-autophagic pool contributing to heterogeneity. The capacity of a network to yield a depolarized daughter mitochondrion by a fission event is fundamental to the generation of heterogeneity. Several studies and data presented here provide a potential explanation, suggesting that protein and membranous structures are unevenly distributed within the individual mitochondrion and that inner membrane components do not mix during a fusion event to the same extent as the matrix components do. In conclusion, mitochondrial subcellular heterogeneity is a reflection of the mitochondrial lifecycle that involves frequent fusion events in which components may be unevenly mixed and followed by fission events generating disparate daughter mitochondria, some of which may fuse again, others will remain solitary and join a pre-autophagic pool. PMID- 19549574 TI - Advances in antibacterial therapy against emerging bacterial pathogens. AB - During the last decade, both gram-positive and gram-negative bacteria that are resistant to most or all available antibacterial classes have become increasingly prevalent nosocomial pathogens, particularly among immunocompromised patients and those hospitalized in intensive care units. Among gram-positive bacteria, increasing concerns are posed for health care- and community-associated methicillin-resistant Staphylococcus aureus (MRSA), S aureus with reduced susceptibility to vancomycin, and vancomycin-resistant enterococci (VRE). A spectrum of newer antibacterial agents has been developed for the treatment of multi-resistant gram-positive bacteria, such as linezolid, tigecycline, daptomycin, and novel glycopeptides. Gram-negative bacteria have also developed multidrug resistance (MDR), which in the Enterobacteriacae is commonly due to the production of extended-spectrum beta-lactamases and carbapenemases of VIM, IMP, or KPC types. Currently, non-fermenting bacteria such as Pseudomonas aeruginosa and Acinetobacter baumannii are commonly resistant to all available antibiotics, including the newer agents. Colistin retains activity against most P aeruginosa and A baumannii, but its clinical use remains questionable, while newer carbapenems and tigecycline have limited additional advantages. Rational use of newer antibacterial agents coupled with enhanced infection control measures may be able to sufficiently control MDR organisms as to allow hematological patients to recover from serious infectious complications. PMID- 19549575 TI - Recent advances in antifungal prevention and treatment. AB - Invasive fungal infections are important causes of morbidity and mortality in patients with bone marrow failure syndromes and hematological malignancies, or who are undergoing allogeneic hematopoietic stem cell transplantation (HSCT). Current epidemiological trends indicate a shift toward infections by Aspergillus spp., non-albicans Candida spp., and previously uncommon fungal pathogens that have decreased susceptibility to the available antifungal agents. The last two decades have seen substantial improvements in the clinical, laboratory, and radiological diagnosis of these infections and the development of new antifungal compounds. Progress has been made in establishing disease definitions and paradigms for antifungal intervention and in the design and conduct of interventional clinical trials. Collectively, these advances have led to major but ongoing changes in the management of patients at risk of or being affected by invasive fungal infections. This article reviews current approaches to prevention and treatment of opportunistic fungal infections in immunocompromised patients with hematological disorders and discusses novel approaches to antifungal chemotherapy and adjunctive treatments. PMID- 19549576 TI - Antiviral therapy in patients with hematologic malignancies, transplantation, and aplastic anemia. AB - Advances in supportive care over the past two decades have decreased the morbidity and mortality attributed to opportunistic infections in immunocompromised patients, including those with hematologic malignancies, hematopoietic stem cell transplantation (HSCT), and aplastic anemia. Despite advances in antiviral therapy, opportunistic viral infections such as influenza, herpes simplex virus (HSV), varicella zoster virus (VZV), and cytomegalovirus (CMV) still cause significant morbidity and mortality in patients with compromised host defenses. Antiviral agents are key antimicrobials used for treatment and prophylaxis of viral infections in immunocompromised hosts. Currently, there are more than 40 antiviral agents approved for clinical use, but the majority of these agents are for the treatment of human immunodeficiency virus (HIV) or viral hepatitis. This review will focus on antiviral agents used for the treatment of herpesviruses (HSV, VZV, CMV), community-acquired respiratory viruses (influenza), and adenoviruses. Antiviral agents used for the treatment of HIV and viral hepatitis will not be addressed in this review. PMID- 19549577 TI - Recent advances in diagnostic microbiology. AB - The past decade has seen a surge in the development of a variety of molecular diagnostics designed to rapidly identify or characterize medically important microorganisms. We briefly review important advances in molecular microbiology, and then discuss specific assays that have been implemented in clinical microbiology laboratories throughout the country. We also discuss emerging methods and technologies that will soon be more widely used for the prompt and accurate detection of the agents of infectious diseases. PMID- 19549578 TI - Neutropenic fever syndromes in patients undergoing cytotoxic therapy for acute leukemia and myelodysplastic syndromes. AB - Fever represents the major surrogate of infection in neutropenic cancer patients. A number of neutropenic fever syndromes have been recognized, the causes and significance of which will vary depending upon the clinical context. First neutropenic fever syndromes are typically of bacterial origin, the character of which may be influenced by whether antibacterial chemoprophylaxis has been administered. Persistent neutropenic fevers are documented during the empirical systemic antibacterial therapy for the first neutropenic fever, the cause of which is likely outside the spectrum of activity of the initial therapy. Recrudescent neutropenic fevers, defined by the appearance of a new fever after defervescence of the first fever, are often a function of invasive fungal infection or gram-positive infections outside the spectrum of the initial empirical antibacterial regimen. The myeloid reconstitution syndrome occurs in parallel with neutrophil recovery from aplasia and may not necessarily represent new infection. Recognition of these patterns can help the clinician make better clinical judgments and management plans. PMID- 19549579 TI - Infections in patients with aplastic anemia. AB - Infection is a major cause of death in patients with aplastic anemia (AA). There are differences between the immunocompromised state of a patient with AA and the patient who is neutropenic due to chemotherapy and this leads to a difference in the infections that they incur. Prolonged neutropenia is one of the largest risk factors for the development of infections with the invasive mycoses and bacteria. Recovery from neutropenia is directly related to survival, and supportive care plays a large role in protection while the patient is in a neutropenic state. The most common invasive mycoses include the Aspergillus species, Zygomycetes, Candida spp., and Fusarium spp. Bacterial infections that are seen in patients with AA include gram-positive coagulase-negative Staphylococcus species, Enterococcus, Staphylococus aureus, Clostridium spp., Micrococcus, alpha hemolytic streptococci, Listeria monocytogenes, and Bacillus cereus. Gram negative infections including gram-negative bacilli, Escherichia coli, Salmonella, Bacteroides fragilis, Klebsiella oxytoca, Klebsiella pneumonia, Aeromonas hydrophilia, Pseudomonas aeruginosa, and Vibrio vulnificus. Viral infections are much less common but include those that belong to the Herpesviridae family, community-acquired respiratory viral infection, and the viral hepatitides A, B, and C. Evidence of the parasite Strongyloides stercoralis has also been documented. This review discusses the major invasive fungal infections, bacterial pathogens, parasites, and viral infections that are found in patients with AA who are treated with immunosuppressive therapy. The specific immune impairment and current treatment parameters for each of these classes of infection will also be discussed. PMID- 19549580 TI - Infections in patients with multiple myeloma. AB - Infectious diseases are the leading causes of death in patients with multiple myeloma (MM). Major changes have occurred in the spectrum of infections paralleling the changes in the treatment of MM. Bacteria (particularly gram negative organisms) are the most frequent etiologic agents, and invasive fungal infections caused by molds (Aspergillus sp. and Fusarium sp.) have been increasingly reported. New infections have emerged with an increase in the intensity of the treatment of MM. Management of infections in MM patients must include recognition of the likely pathogens. Specific strategies of diagnosis, prophylaxis, and empirical and specific therapy are based on host factors and probable pathogens. PMID- 19549582 TI - Infections in pediatric patients with hematologic malignancies. AB - Despite significant advances in supportive care, infection remains second only to malignancy as a cause of death in pediatric oncology patients, and infection accounts for a large fraction of treatment-related costs. Multiple risk factors contribute to infection-related morbidity, chief among them the immunosuppressive effects of leukemia itself and of cytotoxic chemotherapy, prolonged hospitalization and antibiotic use, and loss of barrier integrity associated with mucositis and the need for indwelling central access. While viruses are the most common causes of infection, bacteria are responsible for most life-threatening complications. Gram-negative bacilli are a concern for all patients undergoing treatment, while a subset of gram-positive organisms, particularly viridans streptococci, become significant pathogens in children receiving profoundly immunosuppressive therapy. Invasive fungal infections are also a serious risk for morbidity and mortality in this population. Availability of new antimicrobial agents has made it possible to treat infectious complications more effectively, but their availability is also leading to an increased prevalence of highly resistant pathogens. Future work in pediatric oncology will need to include measures to reduce the immunosuppressive effects of anti-cancer therapy, provide targeted treatment for infections, and better identify groups of patients at high risk for infectious complications, who may benefit from antimicrobial prophylaxis or more aggressive empirical therapy. PMID- 19549581 TI - Management of infections complicating allogeneic hematopoietic stem cell transplantation. AB - The use of allogeneic hematopoietic stem cell transplantation for the treatment of hematologic malignancies, as well as some benign hematologic disorders, has continued to grow over the last 10 years. The availability of this procedure to an increasing number of patients has been facilitated by the use of newer techniques, including reduced intensity conditioning (RIC) regimens, peripheral blood stem cells (PBSCs) and cord blood as donor sources, graft manipulation such as selective T-cell depletion, and other in vitro and in vivo attempts to reduce the risk and severity of graft-versus-host disease (GVHD) after transplantation without losing the potential benefits of a graft-versus-tumor effect for patients with hematologic malignancies. The underlying theme of many of these newer techniques has been to minimize the severity and duration of transplant-related immune suppression, thus reducing the risk of morbidity and mortality from infectious complications. This article reviews immune suppression and recovery that occur after allogeneic stem cell transplantation, with changes in the epidemiology, and some of the recent advances that have been made in management of infectious complications. PMID- 19549583 TI - In vitro and in vivo pharmacokinetic characterization of two novel prodrugs of zidovudine. AB - This work deals with the in vitro and in vivo pharmacokinetic characterization of 3'-azido-2',3'-dideoxy-5'-O-oxalatoylthymidine (AZT-Ac) and 3'-azido-2',3' dideoxy-5'-O-isonicotinoyl-thymidine (AZT-Iso), two novel prodrugs of the anti HIV agent zidovudine [3'-azido-2',3'-dideoxythymidine (AZT)]. AZT, AZT-Ac and AZT Iso intestinal permeation properties and plasma concentration profiles in rats after intravenous administration were studied. Using the everted gut sac intestinal permeation assay, it was observed that AZT was subjected to saturable transport mechanisms in the jejunum and the proximal ileum, while no saturation was found in the distal ileum. AZT-Ac was able to permeate the intestinal segment at a lower rate than AZT but resisting enzymatic hydrolysis, while no evidence of saturation was found. On the other hand, AZT-Iso was completely hydrolyzed in the intestinal tissue, with AZT being found in the permeated samples. In vivo studies demonstrated that AZT plasma half-life (t(1/2)) is extended after administration of AZT-Ac compared to AZT (2.16 and 0.96h, respectively), while after administering AZT-Iso the t(1/2) of the regenerated AZT was shorter (0.38h). A relationship is proposed between these observed in vivo pharmacokinetic features and previous studies of protein-binding properties, concluding that AZT-Ac is a very promising prodrug of AZT in the search for more effective and safer anti-HIV agents. PMID- 19549584 TI - Identification of bisindolylmaleimides and indolocarbazoles as inhibitors of HCV replication by tube-capture-RT-PCR. AB - We devised a screening method for hepatitis C virus (HCV) inhibitors by exploiting the JFH1 viral culture system. The viral RNA released in the medium was adsorbed onto PCR plates, and real-time RT-PCR was performed by directly adding the one-step RT-PCR reaction mixture to the wells. The "tube-capture-RT PCR" method obviates the need for labor-intensive RNA isolation and should allow high-throughput screening of HCV inhibitors. To substantiate the validity of the assay for drug screening, a pilot screen of an inhibitor library composed of 95 compounds was performed. In addition to the known inhibitors of HCV replication included in the library, the assay identified the PKC inhibitor bisindolylmaleimide I (BIM I) as an HCV replication inhibitor. BIM I was also effective in reducing the viral protein level in genotype 1b and 2a subgenomic replicon cells, indicating inhibition of HCV replication. Further assays revealed that a broad range of bisindolylmaleimides and indolocarbazoles inhibit HCV, but no correlation was found between the PKC inhibition pattern and anti-HCV activity. These series of compounds represent new classes of inhibitors that may warrant further development. PMID- 19549585 TI - Genotypic analysis of the protease and reverse transcriptase of non-B HIV type 1 clinical isolates from naive and treated subjects. AB - One hundred and ninety-two pol sequences of drug-naive and drug-experienced subjects infected with non-B HIV-1 subtypes were analyzed to identify treatment related amino acid changes which might be relevant for drug-resistance and possibly not included in the accepted mutation list for the B subtype. The correspondence analysis identified non-B-specific and subtype-specific polymorphisms which should not be mistaken for mutations. Multiple chi(2) were performed to detect the differences between naive vs treated subjects and between different subtypes. To verify the contribution of each single mutation to the resistance levels as predicted by the Virtual Phenotype-LM, simple univariate linear regression was used with fold resistance as a dependent variable and individual mutations as predictors. Commonly accepted protease (PR) and reverse transcriptase (RT) positions along with mutants at RT positions 118 and 90 were significantly associated with treatment. Two unusual PR (K14R and I66F) and five RT positions (E28K, S68G, H221Y, L228R/H and P294A) were also associated with treatment (p<0.01). Only minimal variations were observed with respect to commonly accepted amino acid changes. All amino acid changes correlated with treatment influenced the resistance levels to each single drug. Our findings demonstrate that there are no substantial differences regarding known resistance associated mutations and the newly emergent substitutions between non-B and B subtype strains. PMID- 19549586 TI - Membrane-initiated actions of estrogen on the endothelium. AB - Estrogen-induced rapid, membrane-initiated activation of numerous signal transduction cascades has been shown in animal, cellular and molecular vascular studies, which support the favorable effects of estrogen on vascular structure and function. These effects are mediated by distinct forms of estrogen receptor (ER) alpha. This includes estrogen-stimulated, rapid activation of endothelial nitric oxide synthase (eNOS), resulting in elaboration of the athero-protective, angiogenesis-promoting product nitric oxide (NO). An N-terminus truncated short isoform of ERalpha, ER46, plays a critical role in membrane-initiated, rapid responses to 17beta-estradiol (E2) in human endothelial cells (ECs). We have proposed a ER46-centered, eNOS-activating molecular complex in human EC caveolar membranes, containing c-Src, phosphatidylinositol 3-kinase (PI3K), Akt and eNOS. In this review, we describe estrogen-induced, rapid, non-genomic actions in the endothelium. PMID- 19549589 TI - Signaling-dependent nuclear export of estradiol receptor controls cell cycle progression in breast cancer cells. AB - Estradiol receptor plays a key role in breast cancer and specific hormonal therapies blocking the receptor functions have been developed. Unfortunately, many patients become resistant to this treatment and develop metastatic breast tumors. The causes of breast tumor progression and hormonal therapy resistance are still debated. Many proteins are mislocalized in human cancers, and increasing evidence indicates that nuclear exclusion of estradiol receptor is involved in tumorigenesis of breast cancer cells and hormonal therapy resistance. Therefore, analysis of intracellular localization of estradiol receptor together with screening for specific compounds that redirect the mislocalized receptor to the correct subcellular compartment is a very promising approach to the discovery of novel anticancer compounds. We recently dissected estradiol receptor nuclear export in breast cancer cells and its dependence on PI3-K. This export has a strong impact on cell cycle progression. A peptide mimicking the nuclear export sequence of estradiol receptor specifically traps the receptor in nuclear compartment and blocks the S-phase entry of target cells. PMID- 19549587 TI - Non-genomic regulation of vascular cell function and growth by estrogen. AB - Estrogens exert rapid, non-genomic effects, which are mediated by plasma membrane associated estrogen receptors (mER) mERalpha and mERbeta, and the intracellular transmembrane G protein-coupled estrogen receptor (GPER). Membrane-initiated responses contribute to transcriptional activation, resulting in a complex interplay of nuclear and extra-nuclear mechanisms that mediate the acute physiological responses to estrogens. Non-genomic estrogen signaling also activates a variety of intracellular estrogen signaling pathways that regulate vascular function and cell growth involving rapid but also long-term effects. This review discusses recent advances in understanding of the mechanisms of non genomic estrogen receptor signaling in the vascular wall. PMID- 19549590 TI - Nongenomic activation of spermatozoa by steroid hormones: facts and fictions. AB - The rapid effects of steroids on spermatozoa have been demonstrated for the first time two decades ago. Progesterone (P), which is present throughout the female genital tract with peaks of levels in the cumulus matrix surrounding the oocyte, stimulates several sperm functions, including hyperactivation and acrosome reaction. These effects are mediated by an extranuclear pathway, as P stimulates an influx of calcium, the tyrosine phosphorylation of sperm proteins and other signalling cascades in a rapid manner. Whether these effects are receptor mediated and which receptors mediate these effects are still a matter of discussion despite all the efforts of the scientific community aimed at identifying them during the last 20 years. Although responsiveness to P is related to sperm fertilizing ability, the physiological role of P during the process of fertilization is discussed, and recent evidence points for a role of the steroid as a chemotactic agent for sperm. A similar situation applies for estrogens (E), which have been shown to induce direct effects on sperm by an extranuclear pathway. In particular, E appear to decrease acrosome reaction in response to P, exerting a role in ensuring an appropriate timing for sperm exocytosis during the process of fertilization. PMID- 19549592 TI - New aspects of rapid aldosterone signaling. AB - Aldosterone, the endogenous ligand of the mineralocorticoid receptor (MR) in humans, is a steroid hormone that regulates salt and water homeostasis. Recently, additional pathophysiological effects in the renocardiovascular system have been identified. Besides genomic effects mediated by activated MR, rapid aldosterone actions that are independent of translation and transcription have been documented. While these nongenomic actions influence electrolyte homeostasis, pH and cell volume in classical MR target organs, they also participate in pathophysiological effects in the renocardiovascular system causing endothelial dysfunction, inflammation and remodeling. The mechanisms conveying these rapid effects consist of a multitude of signaling molecules and include a cross-talk with genomic aldosterone effects as well as with angiotensin II and epidermal growth factor receptor signaling. Rapid corticosteroid signaling via the MR has also been demonstrated in the brain. Altogether, the function of nongenomic aldosterone effects seems to be to modulate other signaling cascades, depending on the surrounding milieu. PMID- 19549588 TI - Control of CNS neuronal excitability by estrogens via membrane-initiated signaling. AB - It is well known that many of the actions of 17beta-estradiol (E2) in the central nervous system (CNS) are mediated via intracellular receptor/transcription factors that interact with steroid response elements on target genes. However, there is compelling evidence for membrane-associated steroid receptors for E2 in hypothalamic and other brain neurons. Indeed, we are just beginning to understand how E2 signals via membrane receptors, and how these signals impact not only membrane excitability but also gene transcription in neurons. We know that E2 can rapidly alter neuronal activity within seconds, indicating that some cellular effects can occur via membrane-delimited events. In addition, E2 can affect second messenger systems including calcium mobilization and a plethora of kinases to alter cell signaling. This review will concentrate on rapid membrane-initiated and intracellular signaling by E2 in the hypothalamus and hippocampus, the nature of receptors involved and how they contribute to CNS functions. PMID- 19549591 TI - Signaling inputs to progesterone receptor gene regulation and promoter selectivity. AB - Progesterone receptors (PR) select and control genetic programs in the breast during normal mammary gland development, and progestin-driven processes contribute to the initiation and/or progression of breast cancer [Beral, V., 2003. Breast cancer and hormone-replacement therapy in the Million Women Study. Lancet 362, 419-427; Chlebowski, R.T., Hendrix, S.L., Langer, R.D., Stefanick, M.L., Gass, M., Lane, D., Rodabough, R.J., Gilligan, M.A., Cyr, M.G., Thomson, C.A., et al., 2003. Influence of estrogen plus progestin on breast cancer and mammography in healthy postmenopausal women: the Women's Health Initiative Randomized Trial. JAMA 289, 3243-3253]. Throughout the mammalian life span, progesterone exerts varying biological consequences on the mammary epithelial compartment, from brief proliferative spurts that occur with each luteal phase of the menstrual cycle to the massive expansion of the pregnant gland in preparation for lactation [Brisken, C., Park, S., Vass, T., Lydon, J.P., O'Malley, B.W., Weinberg, R.A., 1998. A paracrine role for the epithelial progesterone receptor in mammary gland development. Proc. Natl. Acad. Sci. U.S.A. 95, 5076-5081; Ismail, P.M., Amato, P., Soyal, S.M., DeMayo, F.J., Conneely, O.M., O'Malley, B.W., Lydon, J.P., 2003. Progesterone involvement in breast development and tumorigenesis-as revealed by progesterone receptor "knockout" and "knockin" mouse models. Steroids 68, 779-787]. These processes, while important developmentally, can become deregulated in breast cancer, thereby contributing to unchecked proliferation, increased survival, and invasive behaviors. Recently, our lab has focused on the molecular mechanisms, including phosphorylation events, by which PRs select specific target genes in response to progestins and other mitogenic hormonal signals (i.e. EGF, heregulin). Herein, we discuss the actions of cytoplasmic signaling molecules such as c-Src and mitogen-activated protein kinases as key mediators of PR promoter selectivity. PMID- 19549594 TI - Evolva breeds small molecule drugs au naturel. PMID- 19549595 TI - Predictable enzymatic glycosylation. AB - An innovative approach for manipulating glycosyltransferase-catalyzed glycosylation has now been developed (Truman et al.). Created using a domain swapping strategy, these chimeric glycotransferases have predictable substrate specificity and may lead to the breakthrough developments in the preparation of carbohydrate-containing molecules of biological interest. PMID- 19549593 TI - Novel non-genomic signaling of thyroid hormone receptors in thyroid carcinogenesis. AB - The thyroid hormone receptors (TRs) are transcription factors that mediate the pleiotropic activities of the thyroid hormone, T3. Four T3-binding isoforms, TRalpha1, TRbeta1, TRbeta2, and TRbeta3, are encoded by two genes, THRA and THRB. Mutations and altered expression of TRs have been reported in human cancers. A targeted germ-line mutation of the Thrbeta gene in the mouse leads to spontaneous development of follicular thyroid carcinoma (TRbeta(PV/PV) mouse). The TRbetaPV mutant has lost T3-binding activity and displays potent dominant negative activity. The striking phenotype of thyroid cancer exhibited by TRbeta(PV/PV) mice has recently led to the discovery of novel non-genomic actions of TRbetaPV that contribute to thyroid carcinogenesis. These actions involve direct physical interaction of TRbetaPV with cellular proteins, namely the regulatory subunit of the phosphatidylinositol 3-kinase (p85alpha), the pituitary tumor transforming gene (PTTG) and beta-catenin, that are critically involved in cell proliferation, motility, migration, and metastasis. Thus, a TRbeta mutant (TRbetaPV), via a novel mode of non-genomic action, acts as an oncogene in thyroid carcinogenesis. PMID- 19549596 TI - Oxygenase catalyzed 5-methylcytosine hydroxylation. AB - Recent reports identify the oxygenase catalyzed production of 5 hydroxymethylcytosine as a modification to mammalian DNA (Kriaucionis and Heintz, 2009; Tahiliani et al., 2009). This discovery has potential far-reaching implications for epigenetic regulation and will stimulate efforts to identify new nucleic acid modifications. PMID- 19549597 TI - An FAD-dependent pyridine nucleotide-disulfide oxidoreductase is involved in disulfide bond formation in FK228 anticancer depsipeptide. AB - Disulfide bonds are rare in bacterial natural products, and the mechanism of disulfide bond formation in those products is unknown. Here we characterize a gene and its product critical for a disulfide bond formation in FK228 anticancer depsipeptide in Chromobacterium violaceum. Deletion of depH drastically reduced FK228 production, whereas complementation of the depH-deletion mutant with a copy of depH on a medium copy-number plasmid not only fully restored the FK228 production but also significantly increased the FK228 yield. Purified 6xHis tagged DepH fusion protein in native form is a homodimer of 71.0 kDa, with each monomer containing one molecule of FAD. DepH efficiently converts an immediate FK228 precursor to FK228 in the presence of NADP(+). We conclude that DepH is an FAD-dependent pyridine nucleotide-disulfide oxidoreductase, specifically and efficiently catalyzing a disulfide bond formation in FK228. PMID- 19549598 TI - Chemical inhibition of a subset of Arabidopsis thaliana GSK3-like kinases activates brassinosteroid signaling. AB - Glycogen synthase kinase 3 (GSK3) is a key regulator in signaling pathways in both animals and plants. Three Arabidopsis thaliana GSK3s are shown to be related to brassinosteroid (BR) signaling. In a phenotype-based compound screen we identified bikinin, a small molecule that activates BR signaling downstream of the BR receptor. Bikinin directly binds the GSK3 BIN2 and acts as an ATP competitor. Furthermore, bikinin inhibits the activity of six other Arabidopsis GSK3s. Genome-wide transcript analyses demonstrate that simultaneous inhibition of seven GSK3s is sufficient to activate BR responses. Our data suggest that GSK3 inhibition is the sole activation mode of BR signaling and argues against GSK3 independent BR responses in Arabidopsis. The opportunity to generate multiple and conditional knockouts in key regulators in the BR signaling pathway by bikinin represents a useful tool to further unravel regulatory mechanisms. PMID- 19549599 TI - Polypeptide translocation by the AAA+ ClpXP protease machine. AB - In the AAA+ ClpXP protease, ClpX uses repeated cycles of ATP hydrolysis to pull native proteins apart and to translocate the denatured polypeptide into ClpP for degradation. Here, we probe polypeptide features important for translocation. ClpXP degrades diverse synthetic peptide substrates despite major differences in side-chain chirality, size, and polarity. Moreover, translocation occurs without a peptide -NH and with 10 methylenes between successive peptide bonds. Pulling on homopolymeric tracts of glycine, proline, and lysine also allows efficient ClpXP degradation of a stably folded protein. Thus, minimal chemical features of a polypeptide chain are sufficient for translocation and protein unfolding by the ClpX machine. These results suggest that the translocation pore of ClpX is highly elastic, allowing interactions with a wide range of chemical groups, a feature likely to be shared by many AAA+ unfoldases. PMID- 19549600 TI - Physically discrete beta-lactamase-type thioesterase catalyzes product release in atrochrysone synthesis by iterative type I polyketide synthase. AB - ATEG_08451 in Aspergillus terreus, here named atrochrysone carboxylic acid synthase (ACAS), is a nonreducing, iterative type I polyketide synthase that contains no thioesterase domain. In vitro, reactions of ACAS with malonyl-CoA yielded a polyketide intermediate, probably attached to its acyl carrier protein (ACP). The addition of ATEG_08450, here named atrochrysone carboxyl ACP thioesterase (ACTE), to the reaction resulted in the release of products derived from atrochrysone carboxylic acid, such as atrochrysone and endocrocin. ACTE, belonging to the beta-lactamase superfamily, thus appears to be a novel type of thioesterase responsible for product release in polyketide biosynthesis. These findings show that ACAS synthesizes the scaffold of atrochrysone carboxylic acid from malonyl-CoA, and that ACTE hydrolyzes the thioester bond between the ACP of ACAS and the intermediate to release atrochrysone carboxylic acid as the reaction product. PMID- 19549601 TI - Substrate-guided design of a potent and selective kallikrein-related peptidase inhibitor for kallikrein 4. AB - Human kallikrein-related peptidase 4 (KLK4/prostase), a trypsin-like serine protease, is a potential target for prostate cancer treatment because of its proteolytic ability to activate many tumorigenic and metastatic pathways including the protease activated receptors (PARs). Currently there are no KLK4 specific small-molecule inhibitors available for therapeutic development. Here we re-engineer the naturally occurring sunflower trypsin inhibitor to selectively block the proteolytic activity of KLK4 and prevent stimulation of PAR activity in a cell-based system. The re-engineered inhibitor was designed using a combination of molecular modeling and sparse matrix substrate screening. PMID- 19549602 TI - Amitriptyline is a TrkA and TrkB receptor agonist that promotes TrkA/TrkB heterodimerization and has potent neurotrophic activity. AB - Neurotrophins, the cognate ligands for the Trk receptors, are homodimers and induce Trk dimerization through a symmetric bivalent mechanism. We report here that amitriptyline, an antidepressant drug, directly binds TrkA and TrkB and triggers their dimerization and activation. Amitriptyline, but not any other tricyclic or selective serotonin reuptake inhibitor antidepressants, promotes TrkA autophosphorylation in primary neurons and induces neurite outgrowth in PC12 cells. Amitriptyline binds the extracellular domain of both TrkA and TrkB and promotes TrkA-TrkB receptor heterodimerization. Truncation of amitriptyline binding motif on TrkA abrogates the receptor dimerization by amitriptyline. Administration of amitriptyline to mice activates both receptors and significantly reduces kainic acid-triggered neuronal cell death. Inhibition of TrkA, but not TrkB, abolishes amitriptyline's neuroprotective effect without impairing its antidepressant activity. Thus, amitriptyline acts as a TrkA and TrkB agonist and possesses marked neurotrophic activity. PMID- 19549603 TI - A cell active chemical GEF inhibitor selectively targets the Trio/RhoG/Rac1 signaling pathway. AB - RhoGEFs (guanine nucleotide exchange factors of the Rho GTPase family) are upstream regulators of cell adhesion and migration pathways, thus representing attractive yet relatively unexplored targets for the development of anti-invasive drugs. We screened for chemical inhibitors of TrioN, the N-terminal GEF domain of the multidomain Trio protein, and identified ITX3 as a nontoxic inhibitor. In transfected mammalian cells, ITX3 blocked TrioN-mediated dorsal membrane ruffling and Rac1 activation while having no effect on GEF337-, Tiam1-, or Vav2-mediated RhoA or Rac1 activation. ITX3 specifically inhibited endogenous TrioN activity, as evidenced by its ability to inhibit neurite outgrowth in nerve growth factor (NGF)-stimulated PC12 cells or C2C12 differentiation into myotubes. This study introduces a selective cell active inhibitor of the Trio/RhoG/Rac1 pathway and validates RhoGEFs as druggable targets. PMID- 19549604 TI - Structural basis for different specificities of acyltransferases associated with the human cytosolic and mitochondrial fatty acid synthases. AB - Animals employ two systems for the de novo biosynthesis of fatty acids: a megasynthase complex in the cytosol (type I) that produces mainly palmitate, and an ensemble of freestanding enzymes in the mitochondria (type II) that produces mainly octanoyl moieties. The acyltransferases responsible for initiation of fatty acid biosynthesis in the two compartments are distinguished by their different substrate specificities: the type I enzyme transfers both the acetyl primer and the malonyl chain extender, whereas the type II enzyme is responsible for translocation of only the malonyl substrate. Crystal structures for the type I and II enzymes, supported by in silico substrate docking studies and mutagenesis experiments that alter their respective specificities, reveal that, although the two enzymes adopt a similar overall fold, subtle differences at their catalytic centers account for their different specificities. PMID- 19549605 TI - Chimeric glycosyltransferases for the generation of hybrid glycopeptides. AB - Glycodiversification, an invaluable tool for generating biochemical diversity, can be catalyzed by glycosyltransferases, which attach activated sugar "donors" onto "acceptor" molecules. However, many glycosyltransferases can tolerate only minor modifications to their native substrates, thus making them unsuitable tools for current glycodiversification strategies. Here we report the production of functional chimeric glycosyltransferases by mixing and matching the N- and C terminal domains of glycopeptide glycosyltransferases. Using this method we have generated hybrid glycopeptides and have demonstrated that domain swapping can result in a predictable switch of substrate specificity, illustrating that N- and C-terminal domains predominantly dictate acceptor and donor specificity, respectively. The determination of the structure of a chimera in complex with a sugar donor analog shows that almost all sugar-glycosyltransferase binding interactions occur in the C-terminal domain. PMID- 19549606 TI - Levamisole is a potential facilitator for the activation of Th1 responses of the subunit HBV vaccination. AB - Chemical compounds activating innate responses may present potential adjuvants for the vaccine development. Levamisole (LMS), demonstrated as a potent adjuvant for DNA and viral killed vaccines in our previous studies, may activate such responses. To confirm this notion, LMS combined with the recombinant HBsAg (rHBsAg) was investigated. Compared to the vaccination with rHBsAg alone, LMS could up-regulate the expressions of TLR7&8, MyD88, IRF7 and their downstream pro inflammatory cytokines including IFN-alpha and TNF-alpha, which promote DCs activation. Strikingly, we find that the combination of LMS and alum adjuvant synergistically enhances immunogenicity of rHBsAg and leads to a robust cell mediated response demonstrated by the higher level of IgG2a/IgG1, T cell proliferation, and importantly, a high level of antigen-specific CTL and IFN gamma production within these activated CD8(+) T cells. The achieved robust responses are at a comparative level with CpG+alum used as a positive control adjuvant in mice. The combination of LMS+alum with rHBsAg may provide a cost effective, safe, and effective therapy to treat those individuals chronically infected by HBV, since antigen-specific cellular immunity is implicated for the clearance of HBV chronic infection. PMID- 19549607 TI - Neutralizing activity and cellular immune responses induced in mice after immunization with apoptotic HIV-1/murine leukemia virus infected cells. AB - Dendritic cells present microbial antigens to T cells after uptake of apoptotic vesicles from infected cells. We previously reported that immunizations with apoptotic HIV-1/murine leukemia virus (MuLV) infected cells lead to induction of both cellular and humoral immune responses as well as resistance to mucosal challenge with live HIV-1/MuLV infected cells. Here we extended those studies and investigated whether apoptotic cells from HIV-1/MuLV infected cells stimulate the production of HIV-1 neutralizing activity. We compared different routes of administration and were able to induce p24- and Nef-specific cellular proliferation after intraperitoneal (i.p.), intranasal (i.n.), subcutaneous (s.c.) and intramuscular (i.m.) immunizations. Serum IgG and IgA antibodies directed against gp160, p24, or Nef were also produced regardless of immunization route used. However, the induction of mucosa-associated IgAs from faeces or vaginal secretions were detected only after either i.p. or i.n. immunizations. We were able to measure neutralizing activity in sera of mice after i.p. and i.n. immunization. Neutralizing reactivity was also detected after s.c. and i.m. immunizations in the presence of the cytokine adjuvant granulocyte macrophage colony stimulating factor (GM-CSF). Conclusively we show induction of cellular and humoral immune responses including neutralizing activity after immunization with apoptotic HIV-1/MuLV infected cells in mice. The results from this study support further evaluations using apoptotic cells as antigen delivery system for vaccination against HIV-1 in other animal models. PMID- 19549608 TI - Vitamin D treatment in multiple sclerosis. AB - Multiple sclerosis is an immune-mediated disease of the central nervous system caused by a complex interplay between several different genetic as well as environmental factors. Vitamin D is one of the most attractive candidates among various environmental factors suggested to be involved in the development of the disease. Epidemiological evidence combined with clinical and laboratory analyses, and experimental animal models, suggest a possible influence of vitamin D on MS susceptibility as well as clinical disease activity. Supplement with vitamin D may reduce the risk of developing MS. An intervention may also reduce the risk of conversion from a first clinical event suggestive of MS to clinical definite MS, as well as reduce the relapse rate among patients with relapsing remitting MS. More studies are, however, needed to determine optimal dose and serum level for vitamin D, as well as target populations and optimal timing for intervention. PMID- 19549612 TI - What's new in Nicotine & Tobacco Research? PMID- 19549614 TI - Detection of testosterone administration based on the carbon isotope ratio profiling of endogenous steroids: international reference populations of professional soccer players. AB - BACKGROUND AND OBJECTIVES: The determination of the carbon isotope ratio in androgen metabolites has been previously shown to be a reliable, direct method to detect testosterone misuse in the context of antidoping testing. Here, the variability in the 13C/12C ratios in urinary steroids in a widely heterogeneous cohort of professional soccer players residing in different countries (Argentina, Italy, Japan, South Africa, Switzerland and Uganda) is examined. METHODS: Carbon isotope ratios of selected androgens in urine specimens were determined using gas chromatography/combustion/isotope ratio mass spectrometry (GC-C-IRMS). RESULTS: Urinary steroids in Italian and Swiss populations were found to be enriched in 13C relative to other groups, reflecting higher consumption of C3 plants in these two countries. Importantly, detection criteria based on the difference in the carbon isotope ratio of androsterone and pregnanediol for each population were found to be well below the established threshold value for positive cases. CONCLUSIONS: The results obtained with the tested diet groups highlight the importance of adapting the criteria if one wishes to increase the sensitivity of exogenous testosterone detection. In addition, confirmatory tests might be rendered more efficient by combining isotope ratio mass spectrometry with refined interpretation criteria for positivity and subject-based profiling of steroids. PMID- 19549613 TI - Identification of GP2, the major zymogen granule membrane glycoprotein, as the autoantigen of pancreatic antibodies in Crohn's disease. AB - BACKGROUND AND AIMS: The aetiopathogenesis of Crohn's disease, an inflammatory bowel disease (IBD), is not yet fully understood. Autoimmune mechanisms are thought to play a role in the development of Crohn's disease, but the target antigens and the underlying pathways have not been sufficiently identified. METHODS: Based on data from immunoblotting and matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry, the major antigenic target of pancreatic autoantibodies (PABs), which are specific for Crohn's disease, was identified. Specificity of autoantibody reactivity was confirmed by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence (IIF) using purified rat and human recombinant GP2 synthesised in transiently transfected mammalian HEK 293 cells. Real-time polymerase chain reaction (rt-PCR) and IIF were used to detect mRNA and antigen localisation in human colon biopsies. RESULTS: The major zymogen granule membrane glycoprotein 2 (GP2) was identified as the autoantigen of PABs in Crohn's disease. PAB-positive sera from patients with Crohn's disease (n = 42) displayed significantly higher IgG reactivity to rat GP2 in ELISA than either PAB-negative sera (n = 31), or sera from patients with ulcerative colitis (n = 49), or sera from blood donors (n = 69) (p<0.0001, respectively). Twenty-eight (66%) and 18 (43%) of 42 PAB-positive sera demonstrated IgG and IgA reactivity to human recombinant GP2 in IIF, respectively. Patients with PAB-negative Crohn's disease (n = 31) were not reactive. GP2 mRNA transcription was significantly higher in colon biopsies from patients with Crohn's disease (n = 4) compared to patients with ulcerative colitis (n = 4) (p = 0.0286). Immunochemical staining confirmed GP2 expression in human colon biopsies from patients with Crohn's disease. CONCLUSION: Anti-GP2 autoantibodies constitute novel Crohn's disease-specific markers, the quantification of which could significantly improve the serological diagnosis of IBD. The expression of GP2 in human enterocytes suggests an important role for anti-GP2 response in the pathogenesis of Crohn's disease. PMID- 19549615 TI - Prolotherapy injections and eccentric loading exercises for painful Achilles tendinosis: a randomised trial. AB - OBJECTIVE: To compare the effectiveness and cost-effectiveness of eccentric loading exercises (ELE) with prolotherapy injections used singly and in combination for painful Achilles tendinosis. DESIGN: A single-blinded randomised clinical trial. The primary outcome measure was the VISA-A questionnaire with a minimum clinically important change (MCIC) of 20 points. SETTING: Five Australian primary care centres. PARTICIPANTS: 43 patients with painful mid-portion Achilles tendinosis commenced and 40 completed treatment protocols. INTERVENTIONS: Participants were randomised to a 12-week program of ELE (n=15), or prolotherapy injections of hypertonic glucose with lignocaine alongside the affected tendon (n=14) or combined treatment (n=14). MAIN OUTCOME MEASUREMENTS: VISA-A, pain, stiffness and limitation of activity scores; treatment costs. RESULTS: At 12 months, proportions achieving the MCIC for VISA-A were 73% for ELE, 79% for prolotherapy and 86% for combined treatment. Mean (95% CI) increases in VISA-A scores at 12 months were 23.7 (15.6 to 31.9) for ELE, 27.5 (12.8 to 42.2) for prolotherapy and 41.1 (29.3 to 52.9) for combined treatment. At 6 weeks and 12 months, these increases were significantly less for ELE than for combined treatment. Compared with ELE, reductions in stiffness and limitation of activity occurred earlier with prolotherapy and reductions in pain, stiffness and limitation of activity occurred earlier with combined treatment. Combined treatment had the lowest incremental cost per additional responder ($A1539) compared with ELE. CONCLUSIONS: For Achilles tendinosis, prolotherapy and particularly ELE combined with prolotherapy give more rapid improvements in symptoms than ELE alone but long-term VISA-A scores are similar. TRIAL REGISTRATION NUMBER: ACTRN: 12606000179538. PMID- 19549616 TI - Validation of the VISA-A questionnaire for Turkish language: the VISA-A-Tr study. AB - OBJECTIVES: To evaluate the validity and reliability of the Turkish version of the Victorian Institute of Sports Assessment-Achilles (VISA-A) questionnaire for patients with Achilles tendinopathy. DESIGN: Fifty-five patients with a diagnosis of Achilles tendinopathy and 55 healthy subjects were included in the study. VISA A questionnaires were translated and culturally adapted into Turkish. The final Turkish version (VISA-A-Tr) was tested for reliability on healthy individuals and patients. Tests for internal consistency, validity and structure were performed on 55 patients. RESULTS: The VISA-A-Tr showed good test-retest reliability (Pearson's r=0.99, p<0.001). The patients with Achilles tendinopathy had a significantly lower score (p<0.001) than the healthy individuals. The VISA-A-Tr score correlated significantly with the Stanish tendon grading system (Spearman's r=-0.86; p<0.001). CONCLUSION: The VISA-A-Tr is a valid and reliable tool for evaluating the severity of Achilles tendinopathy. PMID- 19549617 TI - The Lausanne recommendations: a Dutch experience. AB - OBJECTIVE: This study presents the results of 28 months of preparticipation cardiovascular screening using the Lausanne recommendations, which include a personal and family history, physical examination and electrocardiogram (ECG). DESIGN: From January 2006 to April 2008 the data of the Lausanne screenings carried out at the University Centre of Sports Medicine in Groningen were collected. PARTICIPANTS: 825 cardiovascular screenings were performed of which 397 were excluded. Exclusion criteria were age under 12 or over 35 years, multiple screenings (only the first was included) and known cardiovascular disease. MAIN OUTCOME MEASURES: Negative screening result, (false) positive screening result, medical consumption and number needed to screen. RESULTS: A total of 371 (87%) athletes had a negative screening result. Fifty-five athletes (13%) underwent additional (stage 2) testing and seven (1.6%) further (stage 3) testing. Only 27 athletes (6.3%) were referred for additional testing based only on abnormalities of their ECG. Forty-seven athletes (11%) had a false-positive screening result. Ten athletes (2%) had a positive screening result and three (0.7%) were ultimately restricted from sports participation. Stage 2 medical consumption was 62%, 20% and 18% for one, two and three or more additional tests, respectively. Stage 3 medical consumption was 1.6%. The number of athletes needed to screen to find a single athlete with a potentially lethal cardiovascular disease was 143. CONCLUSION: This study found that when the Lausanne recommendations are implemented in The Netherlands, screening results resemble those found in previous studies. The number of athletes needed to screen to detect one athlete with a potentially lethal cardiovascular disease is also within an acceptable range. PMID- 19549619 TI - Dual antiplatelet therapy for more than 12 months after percutaneous coronary intervention: insights from the Guthrie PCI Registry. AB - OBJECTIVE: To assess the impact of dual antiplatelet (DAP) therapy of >12 months on long-term death and myocardial infarction (MI) after percutaneous coronary intervention (PCI). DESIGN, SETTING AND PATIENTS: Prospective, single-centre, observational study of 1859 consecutive patients who underwent successful PCI of a native coronary artery and survived event-free for at least 12 months. MAIN OUTCOME MEASURES: Combined end point of death or non-fatal MI determined by survival analysis and propensity-adjusted multivariable Cox regression. Similar analyses were performed in the two stent subsets: bare metal stents (n = 835), drug-eluting stents (n = 1024); and three high-risk subsets: diabetic patients (n = 486), patients presenting with MI (n = 713), and those with ACC/AHA type C lesions (n = 717). RESULTS: Baseline characteristics were as follows: mean (SD) age 64 (12) years, male 69%, diabetic 26%, presentation with MI 38%, mean (SD) ejection fraction 49 (12)%, mean (SD) vessel diameter 3.1 (0.5) mm. Duration of DAP was 27 (11) months in "DAP >12 months" and 4.1 (4.1) months in "DAP < or =12 months" (p<0.001). At a median follow-up of 3.4 years after PCI, "DAP >12 months" vs "DAP < or =12 months" had similar incidence of death or MI (9.4% vs 10.3%, log rank p = 0.83). After multivariable adjustment, DAP therapy >12 months was not associated with lower incidence of death or MI than DAP therapy < or =12 months (adjusted HR = 1.01; 95% CI 0.74 to 1.37, p = 0.95). Analysis of each of the five predefined subsets showed similar results. CONCLUSIONS: In patients who undergo successful native coronary PCI and survive event-free for at least 12 months, continuation of dual antiplatelet therapy beyond 12 months does not confer long term protection from death or MI. PMID- 19549618 TI - Synergistic induction of local glucocorticoid generation by inflammatory cytokines and glucocorticoids: implications for inflammation associated bone loss. AB - OBJECTIVES: Synovial fibroblasts and osteoblasts generate active glucocorticoids by means of the 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) enzyme. This activity increases in response to proinflammatory cytokines or glucocorticoids. During inflammatory arthritis synovium and bone are exposed to both these factors. This study hypothesised that glucocorticoids magnify the effects of inflammatory cytokines on local glucocorticoid production in both synovium and bone. METHODS: The effects of inflammatory cytokines (IL 1beta/tumour necrosis factor alpha; TNFalpha) and glucocorticoids, alone or combined, were assessed on the expression and activity of 11beta-HSD1 in primary synovial fibroblasts, primary human osteoblasts and MG-63 osteosarcoma cells. A range of other target genes and cell types were used to examine the specificity of effects. Functional consequences were assessed using IL-6 ELISA. RESULTS: In synovial fibroblasts and osteoblasts, treatment with cytokines or glucocorticoids in isolation induced 11beta-HSD1 expression and activity. However, in combination, 11beta-HSD1 expression, activity and functional consequences were induced synergistically to a level not seen with isolated treatments. This effect was seen in normal skin fibroblasts but not foreskin fibroblasts or adipocytes and was only seen for the 11beta-HSD1 gene. Synergistic induction had functional consequences on IL-6 production. CONCLUSIONS: Combined treatment with inflammatory cytokines and glucocorticoids synergistically induces 11beta-HSD1 expression and activity in synovial fibroblasts and osteoblasts, providing a mechanism by which synovium and bone can interact to enhance anti-inflammatory responses by increasing localised glucocorticoid levels. However, the synergistic induction of 11beta-HSD1 might also cause detrimental glucocorticoid accumulation in bone or surrounding tissues. PMID- 19549620 TI - Vardenafil treatment for patients with pulmonary arterial hypertension: a multicentre, open-label study. AB - BACKGROUND: Vardenafil is a new phosphodiesterase-5 inhibitor that has shown some efficacy in the treatment of pulmonary arterial hypertension (PAH). OBJECTIVE: To examine the long-term effects of vardenafil in patients with PAH. METHODS: A multicentre, open-label study of 1-year's duration was undertaken in 45 patients with PAH to determine the long-term safety and efficacy of vardenafil (5 mg once daily for the first 4 weeks, then 5 mg twice daily) and make a preliminary assessment of its monthly acquisition cost compared with other PAH-active treatments. The patients' clinical features, exercise capacity, WHO functional class and haemodynamic variables were measured at baseline and at 3 and at least 9 months after initiating vardenafil treatment. RESULTS: At the 3 months and a mean (SD) of 14 (3) months (range 9-18) follow-up assessments, the 6 min walking distance was significantly increased from baseline by 70.7 (78.4) m (p<0.001) and 83.4 (91.8) m (p<0.001), respectively. Furthermore, long-term treatment with vardenafil for a mean duration of >1 year was also associated with improvements in haemodynamic parameters, WHO functional class and serum uric acid concentrations. Overall, vardenafil treatment was well tolerated. No patients were withdrawn owing to adverse events and none died during the course of the study. CONCLUSION: Long-term treatment with vardenafil is well tolerated and has sustained beneficial effects on PAH, as measured by patients' exercise capacity, WHO functional class and haemodynamic parameters. PMID- 19549621 TI - Steep left ventricle to aortic root angle and hypertrophic obstructive cardiomyopathy: study of a novel association using three-dimensional multimodality imaging. AB - BACKGROUND: Patients with hypertrophic cardiomyopathy (HCM) exhibit a difference in left ventricular outflow tract (LVOT) obstruction, independently of basal septal thickness (BST). Some patients with HCM have a steeper left ventricle to aortic root angle than controls. OBJECTIVE: To test the predictors of the LV aortic root angle and the association between LV-aortic root angle and LVOT obstruction using three-dimensional imaging. PATIENTS: 153 consecutive patients with HCM (mean (SD) age 46 (14) years, 68% men) and 62 patients with hypertensive heart disease of the elderly (all >65 years of age, 73 (6) years, 34% men) who underwent whole-heart three-dimensional cardiac magnetic resonance (CMR) angiography (1.5 T) and Doppler echocardiography. Forty-two controls (age 43 (11) years, 38% men) who underwent contrast-enhanced multidetector computed tomography and were free of cardiovascular pathology were also studied. MAIN OUTCOMES: LV aortic root angle, BST and maximal non-exercise LVOT gradient were measured in patients with HCM and in hypertensive-elderly patients. Additionally, LV-aortic root angle and BST were measured in controls. RESULTS: The mean (SD) LV-aortic root angle was significantly different (p<0.001) in the three groups: HCM (134 (10) degrees ), hypertensive-elderly (128 (10) degrees ), control (140 (7) degrees ). There was an inverse correlation between age and LV-aortic root angle in the three groups (all p<0.001): HCM (r = -0.56), hypertensive-elderly (r = 0.35), control (r = -0.48). On univariate analysis, in the HCM group, LV-aortic root angle (beta = -0.34, p<0.001), age (beta = 0.23, p = 0.01) and end-systolic volume index (beta = -0.20, p = 0.02), but not BST (beta = 0.02, p = 0.8), were associated with LVOT gradient. On multivariate analysis, only LV-aortic root angle was associated with LVOT gradient. CONCLUSIONS: Patients with HCM have a steeper LV-aortic root angle than controls. In patients with HCM, a steeper LV aortic root angle predicts dynamic LVOT obstruction, independently of BST. PMID- 19549622 TI - Common and dissociable prefrontal loci associated with component mechanisms of analogical reasoning. AB - The ability to draw analogies requires 2 key cognitive processes, relational integration and resolution of interference. The present study aimed to identify the neural correlates of both component processes of analogical reasoning within a single, nonverbal analogy task using event-related functional magnetic resonance imaging. Participants verified whether a visual analogy was true by considering either 1 or 3 relational dimensions. On half of the trials, there was an additional need to resolve interference in order to make a correct judgment. Increase in the number of dimensions to integrate was associated with increased activation in the lateral prefrontal cortex as well as lateral frontal pole in both hemispheres. When there was a need to resolve interference during reasoning, activation increased in the lateral prefrontal cortex but not in the frontal pole. We identified regions in the middle and inferior frontal gyri which were exclusively sensitive to demands on each component process, in addition to a partial overlap between these neural correlates of each component process. These results indicate that analogical reasoning is mediated by the coordination of multiple regions of the prefrontal cortex, of which some are sensitive to demands on only one of these 2 component processes, whereas others are sensitive to both. PMID- 19549623 TI - Improving recruitment to health research in primary care. AB - BACKGROUND: Recruitment to health research is known to be problematic. However, evidence concerning ways of improving recruitment is sparse. OBJECTIVE: To outline the process of recruitment, factors impacting on recruitment success and key areas for further research and development. METHODS: Narrative literature review. RESULTS: This paper argues that three ways of improving recruitment should form the focus of future work: developing a repository of evidence-based techniques and methods which can be introduced by research teams; developing the infrastructure to support recruitment, especially new technologies around the electronic patient record; and increasing public engagement with research, to improve participation by both clinicians and patients. CONCLUSION: Recruitment to health research in primary care remains a major hurdle, and key research and development priorities must be addressed. PMID- 19549625 TI - Symbolism of plants: examples from European-Mediterranean culture presented with biology and history of art: AUGUST: Bittersweet, woody nightshade. PMID- 19549626 TI - Further characterization of ferric-phytosiderophore transporters ZmYS1 and HvYS1 in maize and barley. AB - Roots of some gramineous plants secrete phytosiderophores in response to iron deficiency and take up Fe as a ferric-phytosiderophore complex through the transporter YS1 (Yellow Stripe 1). Here, this transporter in maize (ZmYS1) and barley (HvYS1) was further characterized and compared in terms of expression pattern, diurnal change, and tissue-type specificity of localization. The expression of HvYS1 was specifically induced by Fe deficiency only in barley roots, and increased with the progress of Fe deficiency, whereas ZmYS1 was expressed in maize in the leaf blades and sheaths, crown, and seminal roots, but not in the hypocotyl. HvYS1 expression was not induced by any other metal deficiency. Furthermore, in maize leaf blades, the expression was higher in the young leaf blades showing severe chlorosis than in the old leaf blades showing no chlorosis. The expression of HvYS1 showed a distinct diurnal rhythm, reaching a maximum before the onset of phytosiderophore secretion. In contrast, ZmYS1 did not show such a rhythm in expression. Immunostaining showed that ZmYS1 was localized in the epidermal cells of both crown and lateral roots, with a polar localization at the distal side of the epidermal cells. In maize leaves, ZmYS1 was localized in mesophyll cells, but not epidermal cells. These differences in gene expression pattern and tissue-type specificity of localization suggest that HvYS1 is only involved in primary Fe acquisition by barley roots, whereas ZmYS1 is involved in both primary Fe acquisition and intracellular transport of iron and other metals in maize. PMID- 19549627 TI - The relationship among neuregulin 1-stimulated phosphorylation of AKT, psychosis proneness, and habituation of arousal in nonclinical individuals. AB - BACKGROUND: Previous studies reported an association between weak habituation of skin conductance orienting response and psychosis proneness. The aim of this study was to investigate the relationship among neuregulin 1 (NRG1)-stimulated AKT phosphorylation (a putative marker of psychosis), orienting response habituation, delusional ideas, anxiety, and depression in nonclinical individuals. METHODS: One hundred twenty individuals participated in the skin conductance measurements. Weak and strong habituators were compared on measures of NRG1-stimulated AKT phosphorylation in B lymphoblasts, delusional ideas, anxiety, and depression. The predictors of delusional ideas were determined by multiple regression analysis. RESULTS: Weak habituators displayed higher levels of delusional ideas/anxiety and a lower ratio of phosphorylated AKT as compared with strong habituators. There were 3 significant predictors of delusional ideas: decreased habituation, NRG1-induced AKT phosphorylation, and anxiety. Age, gender, education, IQ, and depression did not predict delusional ideas. CONCLUSIONS: These results suggest that decreased habituation of arousal, NRG1 induced AKT phosphorylation, and anxiety are related to delusional ideation in the general population. PMID- 19549628 TI - Families with mixed eligibility for public coverage: navigating Medicaid, CHIP, and uninsurance. AB - In the midst of health care reform, eligible but uninsured children remain a cause for concern. Children in the same family often have differing eligibility status for public coverage. Mixed eligibility is associated with higher uninsurance rates, even when all children in a family are eligible. Medicaid policies play an important role in creating mixed-eligibility families via age related eligibility thresholds and limited benefits for immigrants; states running separate Children's Health Insurance Program (CHIP) programs have higher uninsurance rates among eligible children. Recent policies to simplify enrollment have not lowered uninsurance among these children. States may improve take-up rates by focusing on eligible children in mixed-eligibility families. PMID- 19549630 TI - Swift: primary data analysis for the Illumina Solexa sequencing platform. AB - MOTIVATION: Primary data analysis methods are of critical importance in second generation DNA sequencing. Improved methods have the potential to increase yield and reduce the error rates. Openly documented analysis tools enable the user to understand the primary data, this is important for the optimization and validity of their scientific work. RESULTS: In this article, we describe Swift, a new tool for performing primary data analysis on the Illumina Solexa Sequencing Platform. Swift is the first tool, outside of the vendors own software, which completes the full analysis process, from raw images through to base calls. As such it provides an alternative to, and independent validation of, the vendor supplied tool. Our results show that Swift is able to increase yield by 13.8%, at comparable error rate. PMID- 19549629 TI - Prestige centrality-based functional outlier detection in gene expression analysis. AB - MOTIVATION: Traditional gene expression analysis techniques capture an average gene expression state across sample replicates. However, the average signal across replicates will not capture activated gene networks in different states across replicates. For example, if a particular gene expression network is activated within a subset or all sample replicates, yet the activation state across the sample replicates differs by the specific genes activated in each replicate, the activation of this network will be washed out by averaging across replicates. This situation is likely to occur in single cell gene expression experiments or in noisy experimental settings where a small sub-population of cells contributes to the gene expression signature of interest. RESULTS AND IMPLEMENTATION: In this light, we developed a novel network-based approach which considers gene expression within each replicate across its entire gene expression profile, and identifies outliers across replicates. The power of this method is demonstrated by its ability to enrich for distant metastasis related genes derived from noisy expression data of CD44+CD24-/low tumor initiating cells. PMID- 19549631 TI - A survey of across-target bioactivity results of small molecules in PubChem. AB - This work provides an analysis of across-target bioactivity results in the screening data deposited in PubChem. Two alternative approaches for grouping related targets are used to examine a compound's across-target bioactivity. This analysis identifies compounds that are selectively active against groups of protein targets that are identical or similar in sequence. This analysis also identifies compounds that are bioactive across unrelated targets. Statistical distributions of compound' across-target selectivity provide a survey to evaluate target specificity of compounds by deriving and analyzing bioactivity profile across a wide range of biological targets for tested small molecules in PubChem. This work enables one to select target specific inhibitors, identify promiscuous compounds and better understand the biological mechanisms of target-small molecule interactions. PMID- 19549632 TI - SOLpro: accurate sequence-based prediction of protein solubility. AB - MOTIVATION: Protein insolubility is a major obstacle for many experimental studies. A sequence-based prediction method able to accurately predict the propensity of a protein to be soluble on overexpression could be used, for instance, to prioritize targets in large-scale proteomics projects and to identify mutations likely to increase the solubility of insoluble proteins. RESULTS: Here, we first curate a large, non-redundant and balanced training set of more than 17 000 proteins. Next, we extract and study 23 groups of features computed directly or predicted (e.g. secondary structure) from the primary sequence. The data and the features are used to train a two-stage support vector machine (SVM) architecture. The resulting predictor, SOLpro, is compared directly with existing methods and shows significant improvement according to standard evaluation metrics, with an overall accuracy of over 74% estimated using multiple runs of 10-fold cross-validation. PMID- 19549633 TI - The ethics of public health practice: balancing private and public interest within tobacco policy. AB - INTRODUCTION: Public health practice is characterized by measuring population health, assessing needs for health care and the provision (directly or indirectly) of services to protect and promote the public's health. It is increasingly explicitly concerned with issues of equity. SOURCES OF DATA: Publications discussing ethical issues in public health. AREAS OF AGREEMENT: Unlike the duties of clinicians, professional standards for public health practice are not well defined. An ethics framework would help the development and implementation of public health policy. AREAS OF CONTROVERSY: Public health strategies have been criticized for being paternalistic and restrictive of personal choice behaviours or for being too pragmatic, and appearing to endorse illegal activities. GROWING POINTS: Historically public health programmes have been delivered at a population level for large groups of people with varying capacity to benefit. Within more autonomy, consumer-orientated political environment, strategy must be more targeted to facilitate healthy choices as defined by the individual. AREAS TIMELY FOR DEVELOPING RESEARCH: Debate is needed on the aims of public health, rights and responsibilities of professionals and citizens and mechanisms for developing and implementing policy. PMID- 19549634 TI - Sports activities after lower limb osteotomy. AB - INTRODUCTION: Active sports participation can be important in some patients with degenerative joint disease in the lower limb. We investigated whether this is possible after an osteotomy for osteoarthritis of the hip, knee and ankle joints. SOURCES OF DATA: We performed a literature search using Medline, Cochrane, CINAHL and Google Scholar with no restriction to time period or language using the keywords: 'osteotomy and sports'. Eleven studies (all level IV evidence) satisfied our inclusion and exclusion criteria. Nine reported on high tibial osteotomies, one on periacetabular osteotomies and one on distal tibial osteotomies. The Coleman Methodology Score to assess the quality of studies showed much heterogeneity in terms of study design, patient characteristics, management methods and outcome assessment. AREAS OF AGREEMENT: Participation in recreational sports is possible in most patients who were active in sports before lower limb osteotomy. In no study were patients able to participate in competitive sports. AREAS OF CONTROVERSY: Intensive participation in sports after osteotomy may adversely affect outcome and lead to failures requiring re operation. GROWING POINTS: Patients may be able to remain active in selected sports activities after a lower limb osteotomy for osteoarthritis. More rapid progression of arthritis is however a possibility. AREAS TIMELY FOR DEVELOPING RESEARCH: Prospective comparative studies investigating activities and sports participation in age-matched patients undergoing osteotomy or joint replacement could lead to useful conclusions. Increased activity and active sports participation may lead to progression of arthritis and earlier failure requiring additional surgery. PMID- 19549635 TI - A cell-autonomous role for WT1 in regulating Sry in vivo. AB - Human patients with Frasier syndrome express reduced levels of the +KTS isoforms of the developmental regulator WT1 and exhibit complete XY gonadal dysgenesis and male-to-female sex reversal. Mice with a targeted mutation that blocks production of these isoforms show a reduction in Sry mRNA in the gonad, but the molecular and cellular basis of this reduction has not been established. Using immunofluorescence analysis, we found a significantly lower level of SRY protein per cell in XY Wt1(+KTS)-null mouse gonads. We also found a reduced number of SRY expressing cells, correlating with a decrease in cell proliferation at and near the coelomic epithelium at 11.5 dpc. No reduction in somatic cell numbers was seen in XX Wt1(+KTS)-null gonads, indicating that the effect of WT1 on cell proliferation is mediated by Sry. Sertoli cell differentiation was blocked in XY Wt1(+KTS)-null mouse gonads, as indicated by the loss of SOX9 and Fgf9 expression, but the addition of recombinant FGF9 to ex vivo gonad cultures rescued the mutant phenotype, as indicated by the induction of the Sertoli-cell specific marker anti-Mullerian hormone. Our data suggest that WT1(+KTS) is involved in the cell-autonomous regulation of Sry expression, which in turn influences cell proliferation and Sertoli cell differentiation via FGF9. Thus, sex reversal in Wt1(+KTS)-null mice and Frasier syndrome patients results from a failure of Sertoli cells both to fully differentiate and to reach sufficient numbers to direct testis development. PMID- 19549636 TI - The disease-protective complement factor H allotypic variant Ile62 shows increased binding affinity for C3b and enhanced cofactor activity. AB - Mutations and polymorphisms in the gene encoding factor H (CFH) have been associated with atypical haemolytic uraemic syndrome, dense deposit disease and age-related macular degeneration. The disease-predisposing CFH variants show a differential association with pathology that has been very useful to unravel critical events in the pathogenesis of one or other disease. In contrast, the factor H (fH)-Ile(62) polymorphism confers strong protection to all three diseases. Using ELISA-based methods and surface plasmon resonance analyses, we show here that the protective fH-Ile(62) variant binds more efficiently to C3b than fH-Val(62) and competes better with factor B in proconvertase formation. Functional analyses demonstrate an increased cofactor activity for fH-Ile(62) in the factor I-mediated cleavage of fluid phase and surface-bound C3b; however, the two fH variants show no differences in decay accelerating activity. From these data, we conclude that the protective effect of the fH-Ile(62) variant is due to its better capacity to bind C3b, inhibit proconvertase formation and catalyze inactivation of fluid-phase and surface-bound C3b. This demonstration of the functional consequences of the fH-Ile(62) polymorphism provides relevant insights into the complement regulatory activities of fH that will be useful in disease prediction and future development of effective therapeutics for disorders caused by complement dysregulation. PMID- 19549641 TI - The seed bank longevity index revisited: limited reliability evident from a burial experiment and database analyses. AB - BACKGROUND AND AIMS: Seed survival in the soil contributes to population persistence and community diversity, creating a need for reliable measures of soil seed bank persistence. Several methods estimate soil seed bank persistence, most of which count seedlings emerging from soil samples. Seasonality, depth distribution and presence (or absence) in vegetation are then used to classify a species' soil seed bank into persistent or transient, often synthesized into a longevity index. This study aims to determine if counts of seedlings from soil samples yield reliable seed bank persistence estimates and if this is correlated to seed production. METHODS: Seeds of 38 annual weeds taken from arable fields were buried in the field and their viability tested by germination and tetrazolium tests at 6 month intervals for 2.5 years. This direct measure of soil seed survival was compared with indirect estimates from the literature, which use seedling emergence from soil samples to determine seed bank persistence. Published databases were used to explore the generality of the influence of reproductive capacity on seed bank persistence estimates from seedling emergence data. KEY RESULTS: There was no relationship between a species' soil seed survival in the burial experiment and its seed bank persistence estimate from published data using seedling emergence from soil samples. The analysis of complementary data from published databases revealed that while seed bank persistence estimates based on seedling emergence from soil samples are generally correlated with seed production, estimates of seed banks from burial experiments are not. CONCLUSIONS: The results can be explained in terms of the seed size-seed number trade-off, which suggests that the higher number of smaller seeds is compensated after germination. Soil seed bank persistence estimates correlated to seed production are therefore not useful for studies on population persistence or community diversity. Confusion of soil seed survival and seed production can be avoided by separate use of soil seed abundance and experimental soil seed survival. PMID- 19549642 TI - Audit of anaesthetist-performed echocardiography on perioperative management decisions for non-cardiac surgery. AB - BACKGROUND: Intraoperative transoesophageal echocardiography is increasingly used for guiding intraoperative management decisions during non-cardiac surgery. Transthoracic echocardiography (TTE) equipment and training is becoming more available to anaesthetists, and its point-of-care application may facilitate real time haemodynamic management and preoperative screening. METHODS: We conducted an audit of transthoracic and transoesophageal echocardiograms, performed by an anaesthetist at a tertiary referral centre over a 9-month period, to identify the effect of echocardiography on clinical decision-making in patients undergoing non cardiac surgery. The indications for echocardiography followed published guidelines. RESULTS: Echocardiographic examinations of 97 patients included 87 transthoracic, and 14 transoesophageal studies. Of 36 studies conducted in the preoperative clinic, eight revealed significant cardiac pathology, necessitating cardiology referral or admission before surgery. Preoperative transthoracic echocardiograms performed on the day of surgery (n=39) led to two cancellations of surgery owing to end-stage cardiac disease, the institution of two unplanned surgical procedures (drainage of pleural and pericardial effusions), and to significant changes in anaesthetic and haemodynamic management, or both in 18 patients. Greater influence on management occurred with emergency surgery (75%) than elective surgery (43%). Intraoperative transthoracic (n=10) and transoesophageal (n=14) echocardiography also altered management (altered surgery in two patients, cancellation in one, and altered haemodynamic management in 18 patients). CONCLUSIONS: Anaesthetist-performed point-of-care TTE and thoracic ultrasound may have a high clinical impact on the perioperative management of patients scheduled for non-cardiac surgery. PMID- 19549644 TI - Pediatric traumatic bronchial rupture; results of early and late presentation. AB - The aim of this study is to evaluate our results in management of bronchial rupture after blunt chest trauma in children either presented early or late and their prognosis. Between January 2000 and December 2007, 25 cases <18 years presented early with blunt chest trauma and underwent surgical treatment of bronchial rupture, and 9 cases presented late. This study included 34 cases with age ranging from 6 to 18 years (mean 11.58+/-2.51 years). Twenty-one of them were males (61.76%) and 13 (38.24%) were females. In early cases, we approached 21 patients (84%) through right thoracotomy and 4 cases (16%) via left thoracotomy, direct sutures of the all early cases (except 3 cases needed lobectomies) were done. Reimplantation after debridment of the edges was done in 4 cases (44.44%) in the late cases, while pulmonary resections were done in 5 cases (55.56%). Mortalities from perioperative cardiac arrest and cerebral hypoxia with failure of resuscitation occurred in 4 patients (11.76%) from all cases. We conclude that early diagnosis and repair of the bronchial rupture gives good results before suppuration of the parenchyma, which leads to pulmonary resection. Meticulous coordination between the surgeon and anaesthiologist is important to prevent mortalities. Keywords: Bronchial rupture; Children; Blunt chest trauma. PMID- 19549643 TI - Influence of patient-controlled i.v. analgesia with opioids on supraventricular arrhythmias after pulmonary resection. AB - BACKGROUND: Postoperative supraventricular arrhythmias (SVA) are common after pulmonary resection and autonomic imbalance is thought to be one of the triggers. Opioids can increase parasympathetic activity and may balance heightened sympathetic tone after operation. We have examined the effect of postoperative patient-controlled analgesia (PCA) with opioids on postoperative SVA. METHODS: Forty-eight patients were randomly assigned to two groups. The GA group received general anaesthesia PCA and PCA with opioids (fentanyl 6 microg ml(-1) and tramadol 3 mg ml(-1)). The GEA group received combined general/epidural anaesthesia plus patient-controlled epidural analgesia (PCEA). Holter recording was completed for 12 h before operation and 12 and 48 h after operation. The incidence of supraventricular tachycardias (SVT), atrial fibrillation, and supraventricular ectopic beats (SVEBs) was evaluated. RESULTS: The incidence of postoperative SVT was significantly lower in the GA group than in the GEA group (3/22 vs 10/22, P=0.021). The incidence of postoperative SVEBs was not statistically different between the groups, but the frequency of postoperative SVEBs increased less in the GA than the GEA group (7/22 vs 15/22, P=0.016). CONCLUSIONS: PCA with opioids (fentanyl and tramadol) can reduce postoperative SVA after pulmonary resection compared with PCEA with ropivacaine. PMID- 19549645 TI - External validation of the modified Thoracoscore in a new thoracic surgery program: prediction of in-hospital mortality. AB - Informed consent prior to any surgical intervention should include in-hospital survival estimation after the procedure performed. The recently developed Thoracoscore predicts well the postoperative mortality possibility. The purpose of our study was to test the modified Thoracoscore performance in our new thoracic program. One hundred and fifty-five consecutive patients underwent thoracic surgery procedure within two years. The procedures performed were: 62 lung resections, 10 open tumor biopsies, 21 neck and mediastinal procedures, 33 chest wall and pleural procedures, 8 tracheal procedures, 3 esophageal procedures, 13 minor cardiac procedures, and 5 chest trauma cases. The modified Thoracoscore was calculated based on the following variables: age, gender, priority of the procedure, malignancy, type of procedure, Zubrod score, ASA class, and number of co-morbidities. The observed mortality was 5.2% (eight deaths) while the predicted one based on the modified Thoracoscore was 4.9%. The scoring system we used had excellent discriminatory ability with a C statistic (0.95, 95% CIs 0.91-0.99). The Hosmer-Lemeshow goodness-of-fit was not statistically significant (P=0.82), indicating acceptable calibration of the model for the present series. The modified Thoracoscore's ability to predict postoperative survival in the whole context of thoracic surgery performs well in our program. Application of any risk scoring system requires external validation and provides comparison of the actual outcomes with other programs. PMID- 19549646 TI - Microalbuminuria and short-term prognosis in patients undergoing cardiac surgery. AB - OBJECTIVES: To examine if preoperative microalbuminuria (MA) is associated with in increased risk of adverse outcomes in patients undergoing elective cardiothoracic surgery, and if adding information on MA could improve the accuracy of the additive EuroSCORE. METHODS: In a follow-up study we included 962 patients undergoing elective cardiothoracic surgery from 1 April 2005 to 30 September 2007 at our department. MA (urine albumin/creatinine ratio between 2.5 25 mg/mmol) was assessed in a morning spot-urine sample. We used population-based medical registries for 30-day follow-up and compared the length of stay and adverse outcomes including (i) all-cause death, myocardial infarction, stroke, or atrial fibrillation, (ii) surgical reintervention, renal insufficiency, sternal wound infection, or septicaemia among patients with and without MA. RESULTS: MA was found in 180 (18.7%) patients. The risk of both combined outcomes (adjusted odds ratios (ORs): 1.00 (95% confidence interval (CI): 0.77-1.30) and 1.18 (95% CI: 0.79-1.75), respectively) and most individual outcomes did not differ between the micro- and normoalbuminuric patients. The patients with MA and an additive EuroSCORE of 5 had a significantly prolonged median length of intensive care unit (ICU) stay (0.15 days [95% CI: 0.04-0.26]) and total hospital stay (0.5 days [95% CI: 0.04-0.96]). Patients with MA had a higher risk of postoperative septicaemia (OR: 12.1 [95% CI: 3.2-45.9]). Area under receiver operating characteristics curves of the EuroSCORE with regard to 30-day mortality was 0.86 both with and without MA. CONCLUSIONS: Preoperative MA in patients undergoing elective cardiothoracic surgery was not associated with most early adverse outcomes. However, risk of septicaemia was higher and patients with MA also had a marginally longer length of ICU and hospital stay. Information on preoperative MA did not improve the accuracy of the additive EuroSCORE. PMID- 19549647 TI - A critical comparison of echocardiographic measurements used for optimizing cardiac resynchronization therapy: stroke distance is best. AB - AIMS: Dyssynchrony assessment in cardiac resynchronization therapy (CRT) is controversial, and there are no standard protocols for optimizing treatment. We studied the feasibility and reproducibility of several echocardiographic measures to optimize CRT pacemaker settings. We also assessed the utility of 'stroke distance' [left ventricular outflow tract velocity-time integral (LVOT VTI)] in performing this function. METHODS AND RESULTS: Thirty patients underwent the following functional assessments; 6 min walk test distance, peak VO(2) consumption on cardiopulmonary exercise testing (VO(2) peak), quality-of-life scoring, and echocardiography; before and at 3 and 6 months after implantation of the CRT device. At 3 months, patients received LVOT VTI-guided optimization of interventricular (VV) and atrioventricular (AV) delays. The feasibility and reproducibility of each optimization measurement was statistically analysed, and the functional benefits of optimization examined. Left ventricular outflow tract VTI, interventricular mechanical delay (IVMD), and tissue Doppler lateral-septal delay showed good feasibility (>90%), whereas LVOT VTI, IVMD, and the 12-segment tissue Doppler dyssynchrony index showed good reproducibility (coefficient of variation <20%). The most feasible and reproducible measure was LVOT VTI. Our optimization protocol necessitated alteration of AV and/or VV delays in 60% of patients at 3 months and was associated with a 50% improvement in functional responder status between 3 and 6 months. CONCLUSION: Left ventricular outflow tract VTI provides us with a single, direct measure of global LV function which is robust, and easily applicable in routine clinical practice, and which is effective at improving response to CRT. PMID- 19549648 TI - A case of mistaken mesial temporal identity. PMID- 19549649 TI - Watching over the medical device industry. PMID- 19549650 TI - Life and Death. It is not wrong to say no. PMID- 19549652 TI - Ireland's recommended colorectal cancer screening programme will not get extra government funding. PMID- 19549654 TI - Whistleblowing is crucial for a "world class" NHS. PMID- 19549655 TI - RCGP supports use of summary care records. PMID- 19549656 TI - Summary care records. No one has asked me. PMID- 19549657 TI - Conservative Party on health. Exit strategy is a vain hope. PMID- 19549658 TI - Conservative Party on health. GP commissioning. PMID- 19549659 TI - Clostridium difficile infection. PMID- 19549660 TI - Paying for expensive medicines. Oncologists and top-ups. PMID- 19549662 TI - Non-communicable diseases. Please redress the balance of millennium development goals. PMID- 19549661 TI - Colorectal cancer. Limitations of trial. PMID- 19549663 TI - Drug hypoglycaemia. Reactive hypoglycaemia in severe mental illness. PMID- 19549666 TI - UK swine flu cases near 3000, as GP leader says use of Tamiflu may need to be rethought. PMID- 19549667 TI - Nursing homes as a reservoir of extended-spectrum beta-lactamase (ESBL)-producing ciprofloxacin-resistant Escherichia coli. AB - BACKGROUND: To assess the prevalence and risk factors for faecal carriage of fluoroquinolone-resistant, extended-spectrum beta-lactamase (ESBL)-producing, Escherichia coli (MDR E. coli) among residents in nursing homes in Northern Ireland. METHODS: Between January 2004 and May 2006, retrospective histories of hospital admissions, antimicrobial treatment and co-morbidities were collected. Faecal samples were cultured for MDR E. coli. These isolates and their ESBL genes were typed by a reference laboratory. RESULTS: Of the 294 patients included in the study, faecal samples from 119 (40.5%) grew MDR E. coli. The proportion of carriers in the different homes ranged from 0% to 75%. Epidemic strain A belonging to the ST131, O25:H4 lineage with the CTX-M-15 enzyme accounted for 58 (49%) of all isolates; its proportion varied from 0% to 100% among homes. Fifty one percent of carriers had no history of recent hospital admission and only 13.5% had a known history of ESBL E. coli colonization or infection. In a multivariate logistic regression model, days of fluoroquinolone use [odds ratio (OR) = 1.33, 95% confidence interval (CI) 1.04-1.69, P = 0.02] and a history of urinary tract infection (OR = 2.56, 95% CI 1.37-4.78, P = 0.003) were the only variables independently associated with the risk of carrying MDR E. coli. CONCLUSIONS: The high level of faecal carriage of MDR E. coli in nursing home residents demonstrates their importance as a reservoir population. Public health measures to combat spread of these organisms should address the needs of this group. PMID- 19549668 TI - Inadvertent non-nucleoside reverse transcriptase inhibitor (NNRTI)-based antiretroviral therapy in dual HIV-1/2 and HIV-2 seropositive West Africans: a retrospective study. PMID- 19549669 TI - Nosocomial spread of ESBL-positive Enterobacter cloacae co-expressing plasmid mediated quinolone resistance Qnr determinants in one hospital in France. PMID- 19549670 TI - Antibiotic use and intussusception in early childhood. AB - OBJECTIVES: Antibiotic use in childhood has been hypothesized to increase the risk of intussusception. Antibiotics have well-known effects on gut motility and microflora-possible mechanisms for the hypothesized association. We evaluated the risk of intussusception according to antibiotic use. METHODS: We conducted a nationwide cohort study of all Danish singleton children born between 1995 and 2003 (n = 611 410) with individual-level information on antibiotic prescriptions, intussusception and potential confounding variables. Using Poisson regression, we estimated rate ratios of intussusception according to antibiotic use, including estimating increases in rate ratios per dose of antibiotics and rate ratios for time periods following antibiotic use. RESULTS: Intussusception was diagnosed in 434 children during 1 180 749 person-years of follow-up. The intussusception rate ratio was 1.51 [95% confidence interval (CI), 1.19-1.91] comparing antibiotic users with non-users. In the first week following use of extended-spectrum penicillins the rate ratio was 4.68 (95% CI, 2.93-7.47). In the first week following use of macrolides the rate ratio was 3.82 (95% CI, 1.22-11.90). The proportion of all cases attributable to extended-spectrum penicillins and macrolides was 4%. CONCLUSIONS: This is the first prospective study to show an association between antibiotics and intussusception. The association was strong, temporal and biologically plausible. The magnitude of the observed effect and a number of sensitivity analyses favour a causal relationship. However, the potential for confounding-by-indication cannot be completely discounted and controlled studies of the observed association will be necessary for more definite confirmation. PMID- 19549671 TI - Establishment of an outpatient and home parenteral antimicrobial therapy service at a London teaching hospital: a case series. AB - BACKGROUND: Outpatient and home parenteral antimicrobial therapy (OHPAT) is becoming increasingly commonplace in the UK, enabling those patients who would previously have been obliged to remain in hospital for intravenous treatment to be managed as outpatients or in their own homes. The OHPAT service at St Mary's Hospital, London, was established in 2004. This paper describes the types of infection, antimicrobial management and outcomes of patients referred to the service in the 3.5 years since its inception. PATIENTS AND METHODS: All inpatients were eligible for OHPAT, provided that they had a serious infection requiring parenteral therapy, were well enough to leave hospital and fulfilled other criteria. We initially used an outpatient clinic model, but as the service developed, treatment was often delivered in patients' homes, with the OHPAT team providing training and assessment of primary care staff. RESULTS: Four hundred and sixty-seven patients were referred to the service between September 2004 and April 2008. Of these, 273 received 303 courses of OHPAT, 48 were discharged on oral therapy and 3 patients declined outpatient therapy; the remaining 143 patients were deemed unsuitable for inclusion, most commonly because the patient was too unwell for discharge (28.7%) or their social situation was inappropriate (14.7%). Causative organisms were identified in two-thirds of cases, with methicillin-resistant Staphylococcus aureus implicated in one-third of these. Mean treatment length was 24 days (range 1-165 days), with 7394 inpatient bed days saved. Less than 5% of patients were readmitted within 28 days with infection- or drug-related problems. There were no cases of Clostridium difficile associated diarrhoea during or after outpatient treatment, despite extensive use of cephalosporins and other broad-spectrum agents. Patients found the service highly satisfactory and felt that it had improved their quality of life during the treatment period. CONCLUSIONS: The introduction of the OHPAT service at St Mary's Hospital has proved to be of benefit to patients and hospital efficiency alike. PMID- 19549672 TI - Anti-acanthamoeba efficacy and toxicity of miltefosine in an organotypic skin equivalent. AB - OBJECTIVES: Acanthamoebae can cause infections of several organs, including eye, skin, lung and brain. Except for Acanthamoeba keratitis, these infections are linked to immunodeficiency. Treatment is generally problematic, due to the lack of sufficiently effective and also easily manageable drugs. In a previous study we discovered that miltefosine (hexadecylphosphocholine) is highly active against Acanthamoeba spp. in vitro. The aim of the current study was to evaluate the suitability of miltefosine for the topical treatment of Acanthamoeba infections. METHODS: Storage life and time dependency, susceptibilities of opportunistic bacterial and fungal pathogens, and synergistic and adverse effects of combinations with other anti-Acanthamoeba substances were determined. Moreover, an organotypic skin equivalent was adapted for investigating the penetration of acanthamoebae into the epidermis and the human tissue tolerability of miltefosine. RESULTS: It was shown that miltefosine can be stored as a 2 mM stock solution and also as a 50 microM dilution over a period of 12 months at 4 degrees C without any loss of activity. Efficacies against staphylococci and Candida albicans were established. Acanthamoebae were able to penetrate the skin equivalent within 24 h. This penetration was prevented by treatment with miltefosine, while miltefosine treatment was well tolerated by the skin equivalent. CONCLUSIONS: Miltefosine has been approved for oral and topical treatment of leishmaniasis and may also be a promising candidate for the topical treatment of Acanthamoeba infections. PMID- 19549673 TI - Validation of a French hospitalized patients' satisfaction questionnaire: the QSH 45. AB - OBJECTIVE: To develop a generic French self-administered instrument for measuring hospitalized patients' satisfaction based on the patient's point of view: the questionnaire for satisfaction of hospitalized (QSH) patients. DESIGN: The development was supervised by a steering committee and undertaken through three standard steps. Item generation was derived from 95 face-to-face interviews, performed in hospitalized patients and in patients scheduled to be admitted. The item reduction led to a 69-item questionnaire. The validation process was based on validity, reliability and some aspects of external validity. SETTING: Medical, surgical and obstetrical departments (n = 187) of public hospitals (n = 11) from different French regions (n = 3). PARTICIPANTS: Eligible patients were adult subjects hospitalized for at least 24 h. MAIN OUTCOME MEASURES: QSH, sociodemographic data, hospitalization department, visual analogue scales of satisfaction. RESULTS: The final version of QSH contained 45 items describing 9 dimensions, leading to 2 composite scores (staff and structure index). The factor structure accounted for 71% of the total variance. Internal consistency was satisfactory (item-internal consistency over 0.40; Cronbach's alpha coefficients ranged from 0.76 to 0.96). The scalability was satisfactory with inlier-sensitive fit (INFIT) statistics inside an acceptable range. Scores of dimensions were strongly positively correlated with visual analogue scale scores (all P < 0.001). External validity showed statistical associations between QSH scores and age or department. Participation rate was 91%. CONCLUSIONS: The availability of a reliable and valid French questionnaire concerning hospitalized patients' satisfaction, exclusively generated from patients' interviews, enables patient feedback to be incorporated in a continuous quality health-care improvement strategy. PMID- 19549674 TI - The assessment of adverse events in hospitals in Brazil. AB - OBJECTIVE: To evaluate the incidence of adverse events in Brazilian hospitals. DESIGN: Retrospective cohort study based on patient record review. SETTING: Three teaching hospitals in the State of Rio de Janeiro, Brazil. PARTICIPANTS: Random sample (1103) of 27 350 adult patients admitted in 2003. Patients under 18 years old, psychiatric patients and patients whose length of stay was less than 24 hr were excluded, and obstetric cases were included. MAIN OUTCOME MEASURE(S): Incidence of patients with adverse events; proportion of preventable adverse events; number of adverse events per 100 patients and incidence density of adverse events per 100 patient-days. RESULTS: The incidence of patients with adverse events was 7.6% (84 of 1103 patients). The overall proportion of preventable adverse events was 66.7% (56 of 84 patients). The incidence density was 0.8 adverse events per 100 patient-days (103 of 13,563 patient-days). The patient's ward was the most frequent location of adverse events (48.5%). In regard to classification, surgical adverse events were the most frequent ones (35.2%). CONCLUSIONS: The incidence of patients with adverse events at the three hospitals was similar to that in international studies. However, the proportion of preventable adverse events was much higher in the Brazilian hospitals. PMID- 19549675 TI - Electro-anatomical mapping in a patient with isolated left ventricular non compaction and left ventricular tachycardia. AB - We report the case of a 59-year-old non-Caucasian man with sustained left ventricular (LV) tachycardia and isolated LV non-compaction. An electro anatomical mapping of the right ventricle and LV with the Carto system was reconstructed. The voltage map excluded the presence of scarred tissue as a possible substrate responsible of the ventricular arrhythmia. PMID- 19549676 TI - Defibrillation testing in patients with atrial fibrillation. AB - Implantable cardioverter defibrillator implantation in patients with atrial fibrillation (AF) is complicated by the need for anticoagulation during defibrillation testing. In this retrospective study, we evaluated the factors associated with successful cardioversion of AF during ventricular defibrillation testing and the safety of our local anticoagulation protocol. PMID- 19549677 TI - Ventricular fibrillation frequency from implanted cardioverter defibrillator devices. AB - AIMS: The dominant frequency (DF) of ventricular fibrillation (VF) provides a measure of cycle length that may relate to the underlying complexity of the arrhythmia. Dominant frequency analysis may therefore provide insights into VF mechanisms, and potentially guide future therapies. Dominant frequency analysis can be undertaken on stored electrograms (EGMs) from implanted cardioverter defibrillator devices (ICDs). Demonstration of the reproducibility of the DF during separate VF events is necessary before using this tool. METHODS AND RESULTS: We identified 82 patients receiving a Medtronic ICD who had two episodes of VF induced during ICD testing. We extracted EGMs recorded during both episodes and determined DF using the fast Fourier transform. The mean DF for the population was 4.7 +/- 0.6 Hz, corresponding to a cycle length of 213 ms. First and second episodes of VF were very highly correlated (interclass correlation = 0.87, P < 0.01) demonstrating that DF was highly reproducible. The 18 patients on Class III agents had a significantly lower DF than the remaining 63 (4.4 +/- 0.4 vs. 4.8 +/- 0.6 Hz, P < 0.01, n = 18). However, the DF of patients with ischaemic heart disease (n = 34) did not differ when compared with dilated cardiomyopathy patients (n = 25) (4.7 +/- 0.6 vs. 4.6 +/- 0.4 Hz, P = 0.3). CONCLUSION: The DF of short intervals of induced VF is highly reproducible and is sensitive to pharmacological interventions that extend effective refractory period. Such estimates of DF may therefore have clinical utility and in patients with ICDs provide a means of investigating mechanisms underlying the initiation and early phases of VF. PMID- 19549678 TI - Atrial overdrive pacing in sleep apnoea: a meta-analysis. AB - AIMS: Sleep apnoea (SA) is a common breathing disorder that affects 5% of the North American adult population. It has been suggested that suppressing periods of bradycardia associated with apnoea may reduce the autonomic imbalance associated with SA, thereby improving the respiratory condition. The goals of this study were to conduct a systematic review to identify all randomized clinical trial data evaluating atrial overdrive pacing (AOP) for the treatment of SA and to perform a meta-analysis to estimate the true effect of AOP on SA. METHODS AND RESULTS: A systematic review of the literature was performed to identify all reports of the effect of AOP for the treatment of SA. To be eligible for analysis, studies had to be randomized and controlled, and use the apnoea hypopnoea index (AHI) to determine the severity of SA. A total of 10 studies were identified, which included 175 patients with intermediate to severe SA. Overall, AOP reduced the AHI by -4.65 episodes/h [95% confidence interval (CI) -8.27 to 1.03, P = 0.01]. In comparison, studies that included a continuous positive airway pressure (CPAP) arm found a greater reduction in AHI with CPAP: -46.3 episodes/h (CI -56.2 to -36.5, P < 0.001). CONCLUSION: Although it appears that AOP is associated with a statistically significant reduction in AHI, the magnitude of this benefit is small and likely not of clinical significance. Atrial overdrive pacing should not be universally indicated in patients with SA, unless they have a conventional indication for cardiac pacing. PMID- 19549680 TI - RAMA1 is a novel kinetochore protein involved in kinetochore-microtubule attachment. AB - During mitosis, kinetochores need to attach to microtubules emanating from spindle poles. Several protein complexes have been shown to mediate the kinetochore-microtubule interaction. However, with the continually growing number of newly identified kinetochore proteins, it is unclear whether all major components of the kinetochore-microtubule interface have been identified. We therefore performed a high-throughput RNAi screen to identify additional factors involved in kinetochore-microtubule attachment, and identified RAMA1 as a novel regulator of this process. Depletion of RAMA1 results in severe chromosome alignment defects and a checkpoint-dependent mitotic arrest. We show that this is due to reduced kinetochore-microtubule attachments. RAMA1 localizes to the spindle and to outer kinetochores throughout all phases of mitosis and is recruited to kinetochores by the core kinetochore-microtubule attachment factor Hec1. Interestingly, unlike Hec1, the association of RAMA1 with kinetochores is highly dynamic, suggesting that it is not a structural component of the kinetochore. Consistent with this, all other kinetochore proteins tested do not require RAMA1 for their kinetochore localization. Taken together, these results identify RAMA1 as a novel kinetochore protein and suggest that RAMA1 may have a direct role in mediating kinetochore-microtubule interactions. PMID- 19549679 TI - Determination of craniofacial growth in patients with untreated Class III malocclusions and anterior crossbites using the centroid method. AB - The aim of this investigation was to assess average Class III craniofacial growth in Japanese males and females, using cross-sectional data from infancy to adulthood. Growth was analysed using the centroid method. Centroid G was geometrically calculated from the triangle Delta abc, which comprised the palatal, articulare-gnathion (Ar-Gn), and A-B planes. Lateral cephalograms were obtained of 323 Japanese males and 611 females aged 2 years or older but under 25 years of age. Each dataset was divided into 2 year intervals according to age and categorized into 10 developmental stages. One-way analysis of variance followed by a Bonferroni's t-test was used to compare the results for each group of males and females separately. No significant gender differences were observed in the palatal plane to Ar-Gn plane angle. In males, significant increases in the palatal plane to A-B angle were found up to 14-15 years of age and in the area of Delta abc, G-G' and Ms-Ms' values up to 16-17 years. Among females, changes in these cephalometric measurements showed tendencies that were nearly identical to those of males. These findings provide the characteristics of Class III malocclusions in the growth and developmental period, and as they can be used as standards for understanding the effectiveness of Class III treatment, they may be useful in orthodontic and/or orthopaedic diagnosis and treatment planning. PMID- 19549681 TI - Direct recruitment of H+-ATPase from lysosomes for phagosomal acidification. AB - The nascent phagosome progressively establishes an acidic milieu by acquiring a proton pump, the vacuolar-type ATPase (V-ATPase). However, the origin of phagosomal V-ATPase remains poorly understood. We found that phagosomes were enriched with the V-ATPase a3 subunit, which also accumulated in late endosomes and lysosomes. We modified the mouse Tcirg1 locus encoding subunit a3, to express an a3-GFP fusion protein. Live-cell imaging and immunofluorescence microscopy revealed that nascent phagosomes received the a3-GFP from tubular structures extending from lysosomes located in the perinuclear region. Macrophages from a3 deficient mice exhibited impaired acidification of phagosomes and delayed digestion of bacteria. These results show that lysosomal V-ATPase is recruited directly to the phagosomes via tubular lysosomes to establish the acidic environment hostile to pathogens. PMID- 19549682 TI - Regulation of beta 4-integrin expression by epigenetic modifications in the mammary gland and during the epithelial-to-mesenchymal transition. AB - The beta 4 integrin is expressed in epithelial cells, a few other cell types and in some carcinomas. Despite this restricted expression pattern and the functional importance of beta 4 integrin in epithelial and carcinoma biology, little is known about how its expression is regulated. Here, we assessed the epigenetic regulation of beta 4 integrin based on the presence of a large CpG island in the beta 4-integrin gene promoter. We separated basal (beta 4+) and luminal (beta 4-) epithelial cells from the mammary glands of K14-eGFP mice and demonstrated that the beta 4-integrin promoter is unmethylated in basal cells and methylated in luminal cells. We also observed that expression of beta 4 integrin and E-cadherin is lost during the epithelial-to-mesenchymal transition (EMT) of mammary gland cells induced by transforming growth factor beta (TGFbeta), which is coincident with de novo DNA methylation, a decrease in active histone modifications (H3K9Ac and H3K4me3) and an increase in the repressive histone modification H3K27me3. Furthermore, TGFbeta withdrawal promotes a mesenchymal-to-epithelial transition (MET) and triggers the re-expression of beta 4 integrin and E-cadherin. Intriguingly, demethylation at either promoter is not obligatory for transcriptional reactivation after TGFbeta withdrawal. However, both H3K9Ac and H3K4me3 modifications are restored during the MET, and H3K27me3 is reduced, strongly suggesting that reversible histone modifications rather than DNA demethylation are the predominant factors in reactivating expression of these genes. Our data indicate that complex epigenetic modifications contribute to the regulation of the beta 4 integrin and E-cadherin. PMID- 19549683 TI - The beta- and gamma-isoforms of type I PIP5K regulate distinct stages of Ca2+ signaling in mast cells. AB - Crosslinking of IgE receptors by antigen initiates Ca2+ mobilization in mast cells by activating phospholipase-C gamma-mediated hydrolysis of phosphatidylinositol-4,5-bisphosphate [PtdIns(4,5)P2]. The resulting inositol 1,4,5-trisphosphate-mediated Ca2+ release from the endoplasmic reticulum (ER) activates store-operated Ca2+ entry, which is necessary for exocytotic release of inflammatory mediators. To investigate roles for PtdIns(4,5)P2-synthesizing isozymes of the type I phosphatidylinositol 4-phosphate 5-kinase family (PIP5K-I) in mast cell signaling, we compared the ectopic expression of wild-type and catalytically inactive PIP5K-I beta in RBL-2H3 mast cells. Surprisingly, both antigen and thapsigargin-stimulated Ca2+ influx were reduced by overexpression of active PIP5K-I beta, whereas antigen-stimulated Ca2+ release from ER stores was unaffected. Consistent with these results, Ca2+ entry stimulated by antigen or thapsigargin was enhanced by expression of a plasma-membrane-associated inositol polyphosphate 5'-phosphatase, whereas antigen-stimulated Ca2+ release from stores was reduced. To investigate the role of PIP5K-I gamma in antigen-stimulated Ca2+ mobilization, we used bone-marrow-derived mast cells from PIP5K-I gamma(-/-) mice. Antigen-stimulated Ca2+ release from ER stores was substantially reduced in the absence of PIP5K-I gamma, but thapsigargin-mediated Ca2+ entry was unaffected. In summary, PIP5K-I gamma positively regulates antigen-stimulated Ca2+ release from ER stores, whereas PIP5K-I beta negatively regulates store operated Ca2+ entry, suggesting that these different PIP5K-I isoforms synthesize functionally distinct pools of PtdIns(4,5)P2 at the plasma membrane. PMID- 19549684 TI - Rescue of atypical protein kinase C in epithelia by the cytoskeleton and Hsp70 family chaperones. AB - Atypical PKC (PKC iota) is a key organizer of cellular asymmetry. Sequential extractions of intestinal cells showed a pool of enzymatically active PKC iota and the chaperone Hsp70.1 attached to the apical cytoskeleton. Pull-down experiments using purified and recombinant proteins showed a complex of Hsp70 and atypical PKC on filamentous keratins. Transgenic animals overexpressing keratin 8 displayed delocalization of Hsp70 and atypical PKC. Two different keratin-null mouse models, as well as keratin-8 knockdown cells in tissue culture, also showed redistribution of Hsp70 and a sharp decrease in the active form of atypical PKC, which was also reduced by Hsp70 knockdown. An in-vitro turn motif rephosphorylation assay indicated that PKC iota is dephosphorylated by prolonged activity. The Triton-soluble fraction could rephosphorylate PKC iota only when supplemented with the cytoskeletal pellet or filamentous highly purified keratins, a function abolished by immunodepletion of Hsp70 but rescued by recombinant Hsp70. We conclude that both filamentous keratins and Hsp70 are required for the rescue rephosphorylation of mature atypical PKC, regulating the subcellular distribution and steady-state levels of active PKC iota. PMID- 19549687 TI - Dynein and mast/orbit/CLASP have antagonistic roles in regulating kinetochore microtubule plus-end dynamics. AB - Establishment and maintenance of the mitotic spindle requires the balanced activity of microtubule-associated proteins and motors. In this study we have addressed how the microtubule plus-end tracking protein mast/orbit/CLASP and cytoplasmic dynein regulate this process in Drosophila melanogaster embryos and S2 cells. We show that mast accumulates at kinetochores early in mitosis, which is followed by a poleward streaming upon microtubule attachment. This leads to a reduction of mast levels at kinetochores during metaphase and anaphase that depends largely on the microtubule minus end-directed motor cytoplasmic dynein. Surprisingly, we also found that co-depletion of dynein rescues spindle bipolarity in mast-depleted cells, while restoring normal microtubule poleward flux. Our results suggest that mast and dynein have antagonistic roles in the local regulation of microtubule plus-end dynamics at kinetochores, which are important for the maintenance of spindle bipolarity and normal spindle length. PMID- 19549685 TI - Caspase cleavage of Atg4D stimulates GABARAP-L1 processing and triggers mitochondrial targeting and apoptosis. AB - Autophagy is an important catabolic process with roles in cell survival and cell death. It sequesters cytosol and organelles within double-membrane autophagosomes that deliver their contents to lysosomes for degradation. Autophagosome biogenesis is coordinated by the autophagy-related protein 4 (Atg4) family of C54 endopeptidases (Atg4A-Atg4D). These enzymes prime and then later delipidate the autophagosome marker, Atg8. Here, we show that one family member, Atg4D, is cleaved by caspase-3 in vitro and in apoptotic cells. Atg4D is a poor priming and delipidation enzyme in vitro, but truncated DeltaN63 Atg4D displays increased activity against the Atg8 paralogue, gamma-aminobutyric acid receptor-associated protein-like 1 (GABARAP-L1). In living cells, DeltaN63 Atg4D stimulates the delipidation of GABARAP-L1, whereas siRNA silencing of the gene expressing Atg4D abrogates GABARAP-L1 autophagosome formation and sensitises cells to starvation and staurosporine-induced cell death. Interestingly, Atg4D overexpression induces apoptosis, which is preceded by the caspase-independent recruitment of Atg4D to mitochondria and is facilitated by a putative C-terminal Bcl-2 homology 3 (BH3) domain. Atg4D also acquires affinity for damaged mitochondria in cells treated with hydrogen peroxide. These data suggest that Atg4D is an autophagy regulator that links mitochondrial dysfunction with apoptosis. PMID- 19549686 TI - Coordinate pathways for nucleotide and EGF signaling in cultured adult neural progenitor cells. AB - The adult subventricular zone (SVZ) contains astrocyte-like stem cells capable of generating new neurons for the olfactory bulb. Adult neurogenesis is driven by a variety of signal systems that can induce synergistic or opposing cellular responses. It is therefore important to gain insight into the underlying downstream signaling pathways. We have previously shown that the nucleotides ADPbetaS and UTP induce rapid Ca2+ transients in cultured SVZ-derived adult neural progenitors and augment growth-factor-mediated progenitor cell proliferation. Here, we investigated signaling pathways elicited by ADPbetaS, UTP and epidermal growth factor (EGF). All three agonists elicit ERK1/2 and CREB phosphorylation but the temporal characteristics differ between the nucleotides and EGF. Differentiation of the progenitors alters the receptor profile. Oligodendrocytes and young neurons, but not astrocytes, lose responsiveness to the agonists. Inhibition experiments are indicative of an ADPbetaS-elicited EGF receptor transactivation. Whereas UTP acts via the P2Y2 receptor, ADPbetaS exerts its function via the P2Y1 receptor and the P2Y13 receptor. Our data demonstrate that nucleotides and EGF induce converging, but also differential, intracellular signaling pathways and suggest that they carry the potential to act synergistically in the control of cell proliferation and cell survival in adult neurogenesis. PMID- 19549688 TI - An adhesion-independent, aPKC-dependent function for cadherins in morphogenetic movements. AB - Cadherin shedding affects migration and occurs in development and cancer progression. By examining the in vivo biological function of the extracellular cadherin domain (CEC1-5) independently of the shedding process itself, we identified a novel function for cadherins in convergent extension (CE) movements in Xenopus. CEC1-5 interfered with CE movements during gastrulation. Unexpectedly, CEC1-5 did not alter cell aggregation or adhesion to cadherin substrates. Instead, gastrulation defects were rescued by a membrane-anchored cadherin cytoplasmic domain, the polarity protein atypical PKC (aPKC) or constitutive active Rac, indicating that CEC1-5 modulates a cadherin-dependent signalling pathway. We found that the cadherin interacts with aPKC and, more importantly, that the extracellular domain alters this association as well as the phosphorylation status of aPKC. This suggests that CE movements require a dynamic regulation of cadherin-aPKC interaction. Our results show that cadherins play a dual role in CE movements: a previously identified adhesive activity and an adhesion-independent function that requires aPKC and Rac, thereby directly connecting cadherins with polarity. Our results also suggest that increased cadherin shedding, often observed in cancer progression, can regulate migration and invasion by modulating polarity protein activity. PMID- 19549689 TI - In vivo imaging reveals a role for Cdc42 in spindle positioning and planar orientation of cell divisions during vertebrate neural tube closure. AB - Specialization of the cell-division process is a common feature of developing embryos, but most studies on vertebrate cell division have focused on cells dividing in culture. Here, we used in vivo four-dimensional confocal microscopy to explore the role of Cdc42 in governing cell division in the developing neural epithelium of Xenopus laevis. We find that Cdc42 is crucial for stable positioning of the metaphase spindle in these cells, but was not required for spindle positioning in epidermal epithelial cells. We also find that divisions in the Xenopus neural plate are planar oriented, and that rotations of mitotic spindles are essential for establishing this orientation. When Cdc42 is disrupted, spindles over-rotate and the final orientation of divisions is changed. Finally, the planar orientation of cell divisions in this tissue seems to be independent of planar cell polarity (PCP) signaling and does not require normal neural morphogenesis. Our data provide new insights into the coordination of cell division and morphogenesis in epithelial cell sheets and reveal novel, cell-type-specific roles for Cdc42 in spindle positioning and spindle orientation. PMID- 19549691 TI - Acetylcysteine and non-ionic isosmolar contrast-induced nephropathy--a randomized controlled study. AB - INTRODUCTION: Intravenous administration of saline and non-ionic isosmolar contrast media significantly reduces the incidence of contrast-induced nephropathy, one of the most common causes of acute renal failure. Results with oral N-acetylcysteine are conflicting. The aim of our study was to evaluate the prophylactic role of N-acetylcysteine in patients with stable chronic renal failure undergoing coronary and/or peripheral angiography and/or angioplasty. METHODS: We randomized 200 elective, consecutive patients (mean age 74.9 +/- 7.3 years; 65% male, 25% diabetics) with basal creatinine clearance 0.5 mg/dl or >25% within 3 days after the procedure. Serum creatinine was measured at baseline, 24, 48 and 72 h after the procedure. RESULTS: Contrast-induced nephropathy was 8/99 (8.1%) in the N-acetylcysteine group versus 6/101 (5.9%) in the placebo group, P = 0.6. No difference was noted in high-risk subgroups such as diabetics (4/25 versus 2/25 P = 0.4) and those with serum creatinine clearance <42.3 ml/min (5/54 versus 4/48; P = 0.9). CONCLUSION: In our experience, N acetylcysteine did not prevent contrast-induced nephropathy in patients receiving isosmolar (iodixanol) contrast media and adequate hydration. PMID- 19549690 TI - Nonprimed and DYRK1A-primed GSK3 beta-phosphorylation sites on MAP1B regulate microtubule dynamics in growing axons. AB - MAP1B is a developmentally regulated microtubule-associated phosphoprotein that regulates microtubule dynamics in growing axons and growth cones. We used mass spectrometry to map 28 phosphorylation sites on MAP1B, and selected for further study a putative primed GSK3 beta site and compared it with two nonprimed GSK3 beta sites that we had previously characterised. We raised a panel of phosphospecific antibodies to these sites on MAP1B and used it to assess the distribution of phosphorylated MAP1B in the developing nervous system. This showed that the nonprimed sites are restricted to growing axons, whereas the primed sites are also expressed in the neuronal cell body. To identify kinases phosphorylating MAP1B, we added kinase inhibitors to cultured embryonic cortical neurons and monitored MAP1B phosphorylation with our panel of phosphospecific antibodies. These experiments identified dual-specificity tyrosine phosphorylation-regulated kinase (DYRK1A) as the kinase that primes sites of GSK3 beta phosphorylation in MAP1B, and we confirmed this by knocking down DYRK1A in cultured embryonic cortical neurons by using shRNA. DYRK1A knockdown compromised neuritogenesis and was associated with alterations in microtubule stability. These experiments demonstrate that MAP1B has DYRK1A-primed and nonprimed GSK3 beta sites that are involved in the regulation of microtubule stability in growing axons. PMID- 19549692 TI - Hypoxia interferes with connective tissue growth factor (CTGF) gene expression in human proximal tubular cell lines. AB - BACKGROUND: Hypoxia plays an important role in kidney injury. By the stabilization of the transcription factor HIF-1, hypoxia affects gene expression also in tubular epithelial cells. Increased expression of connective tissue growth factor (CTGF) is observed in different kidney diseases and is associated with deteriorating renal function. Therefore, we hypothesized that the expression of CTGF might be modulated under hypoxic conditions. METHODS: The human proximal tubular epithelial cell lines HK-2 and HKC-8 were treated with reduced oxygen tension (1% O(2)) or the hypoxia mimetic dimethyloxalyl glycine (DMOG). CTGF was analysed by Western blotting, real-time RT-PCR and luciferase gene expression assays. RESULTS: Exposure of HK-2 or HKC-8 cells to hypoxia or treatment with DMOG for up to 24 h reduced cellular as well as secreted CTGF protein synthesis. Downregulation was also detectable at the mRNA level and was confirmed by reporter gene assays. Hypoxic repression of CTGF synthesis was dependent on HIF 1, as shown by HIF-1alpha knockdown by siRNA. Furthermore, exposure to hypoxia reduced CTGF synthesis in response to TGF-beta. A negative correlation between HIF-1alpha accumulation and CTGF synthesis was also observed in renal cell carcinoma cells (RCC4 and RCC10). Reexpression of von Hippel-Lindau protein reduced HIF-1alpha and increased CTGF synthesis. CONCLUSIONS: We provide evidence that hypoxia inhibits CTGF synthesis in human proximal tubular epithelial cells, involving HIF-1alpha. Under hypoxic conditions, induction of CTGF by TGF-beta was repressed. The reduced synthesis of the profibrotic factor CTGF may contribute to a potential protective effect of hypoxic preconditioning in acute renal injury. PMID- 19549693 TI - Preserving residual renal function in peritoneal dialysis: volume or biocompatibility? PMID- 19549694 TI - Osmomediated natriuresis in humans: the role of vasopressin and tubular calcium sensing. AB - BACKGROUND: The aim was to investigate the unknown mechanism of osmomediated natriuresis. This is the phenomenon by which hypertonic saline (HS) produces a larger natriuresis than isotonic saline (IS), despite the same sodium content. METHODS: Seven healthy volunteers first received HS and then IS (both 3.85 mmol sodium/kg). To investigate the role of calcium metabolism, four patients received HS, two with an activating mutation (ADH) and two with an inactivating mutation (FHH) of the calcium-sensing receptor (CaSR). RESULTS: In healthy volunteers, HS produced mild hypernatraemia, a 4-fold rise in vasopressin (to 2.2 +/- 0.85 pg/mL) and a 3-fold rise in natriuresis compared with a 1.5-fold rise with IS (P = 0.002). This confirmed osmomediated natriuresis. HS caused calciuresis to increase 1.4-fold and then reduced it 1.4-fold, whereas IS failed to increase calciuresis and caused it to fall 3.7-fold (P = 0.05). Natriuresis and calciuresis in ADH patients were similar to healthy volunteers receiving HS, whereas a blunted response was seen in FHH patients. Patient vasopressin levels did not exceed 1.3 pg/mL and changes from baseline were variable. In one FHH patient, a 3-fold rise in vasopressin did not prevent the blunted natriuresis and calciuresis. In one ADH patient, natriuresis and calciuresis were similar to healthy volunteers despite a 1.7-fold fall in vasopressin. CONCLUSIONS: Our data suggest that not only vasopressin (possibly via its V1a receptor), but also the CaSR (which is sensitive to high sodium concentrations) may play a role in osmomediated natriuresis. These results shed new light on osmomediated natriuresis and suggest roles for the CaSR beyond calcium regulation. PMID- 19549695 TI - Polyunsaturated fatty acid (PUFA) in IgA patients. PMID- 19549696 TI - Neutrophil gelatinase-associated lipocalin (NGAL) reflects iron status in haemodialysis patients. AB - BACKGROUND: An iron deficiency is often present in haemodialysis (HD) patients; however, although transferrin saturation (TSAT) of <20% and/or serum ferritin of <200 ng/mL should express iron scarcity, in HD patients high ferritin levels could be related to inflammation rather than reflecting optimal iron stores. METHODS: The aim of the present study was to evaluate serum levels of neutrophil gelatinase-associated lipocalin (NGAL), a small siderophore-binding protein, in a cohort of 56 chronic HD patients in order to determine its possible relationships with iron status. RESULTS: NGAL levels were markedly higher in HD patients than in healthy controls; furthermore, HD patients with TSAT <20% had lower NGAL values than healthy controls, whereas the correction of iron deficiency by means of chronic i.v. iron administration significantly increased NGAL values from baseline. Findings from univariate and multivariate analyses demonstrated that NGAL was a significant predictor of hsCRP, spKT/V and TSAT. In ROC analysis, a NGAL cut-off level of or =50% diameter reduction). Before PCI, perfusion during stress was significantly reduced in regions supplied by stenotic arteries; blood flow velocity (Deltabeta) -3.9 (-9.0 to 0.5) s(-1), perfusion rate (DeltaA x beta) 175.0 (-518.0 to 58.5) s(-1), and refilling time (Deltart) 210 (-22 to 452)ms, compared with the perfusion increase seen in regions supplied by non-stenotic arteries; Deltabeta 1.6 (-0.7 to 4.4) s(-1), DeltaA x beta 151.7 (-67.0 to 300.5) s(-1), and Deltart -47 (-195 to 89) ms, all P < 0.05. At follow-up, regional stress-induced perfusion improved in 29 regions with successful PCI; Deltabeta 0.1 (-2.7 to 3.6), DeltaA x beta 30.5 (-133.3 to 232.1), and Deltart -99 (-247 to 125), all P < or = 0.01, although there was no improvement in nine regions with restenosis; Deltabeta 0.9 (-1.5 to 5.3), DeltaAxbeta 65.7 (-40.8 to 412.6), and Deltart -79 (-268 to 163), P = NS. CONCLUSION: Quantitative CSE has the potential to detect angiographically significant coronary artery stenoses as well as angiographic success after PCI. PMID- 19549700 TI - Quantitative myocardial contrast echocardiography during pharmacological stress for diagnosis of coronary artery disease: a systematic review and meta-analysis of diagnostic accuracy studies. AB - AIMS: We conducted a meta-analysis to evaluate the accuracy of quantitative stress myocardial contrast echocardiography (MCE) in coronary artery disease (CAD). METHODS AND RESULTS: Database search was performed through January 2008. We included studies evaluating accuracy of quantitative stress MCE for detection of CAD compared with coronary angiography or single-photon emission computed tomography (SPECT) and measuring reserve parameters of A, beta, and Abeta. Data from studies were verified and supplemented by the authors of each study. Using random effects meta-analysis, we estimated weighted mean difference (WMD), likelihood ratios (LRs), diagnostic odds ratios (DORs), and summary area under curve (AUC), all with 95% confidence interval (CI). Of 1443 studies, 13 including 627 patients (age range, 38-75 years) and comparing MCE with angiography (n = 10), SPECT (n = 1), or both (n = 2) were eligible. WMD (95% CI) were significantly less in CAD group than no-CAD group: 0.12 (0.06-0.18) (P < 0.001), 1.38 (1.28-1.52) (P < 0.001), and 1.47 (1.18-1.76) (P < 0.001) for A, beta, and Abeta reserves, respectively. Pooled LRs for positive test were 1.33 (1.13-1.57), 3.76 (2.43-5.80), and 3.64 (2.87-4.78) and LRs for negative test were 0.68 (0.55 0.83), 0.30 (0.24-0.38), and 0.27 (0.22-0.34) for A, beta, and Abeta reserves, respectively. Pooled DORs were 2.09 (1.42-3.07), 15.11 (7.90-28.91), and 14.73 (9.61-22.57) and AUCs were 0.637 (0.594-0.677), 0.851 (0.828-0.872), and 0.859 (0.842-0.750) for A, beta, and Abeta reserves, respectively. CONCLUSION: Evidence supports the use of quantitative MCE as a non-invasive test for detection of CAD. Standardizing MCE quantification analysis and adherence to reporting standards for diagnostic tests could enhance the quality of evidence in this field. PMID- 19549702 TI - The use of serum inhibin A and activin A levels in predicting the outcome of 'pregnancies of unknown location'. AB - BACKGROUND: The aim was to evaluate the role of serum inhibin A and activin A in the prediction of the outcome of women with 'pregnancies of unknown location' (PULs). METHODS: Serum human chorionic gonadotrophin (hCG), progesterone, inhibin A and activin A levels were measured at 0 and 48 h. Differences in the mean levels and the change in levels over 48 h expressed as a ratio (48/0 h) were examined between the three outcome groups--failing PUL, intrauterine pregnancy (IUP) or ectopic pregnancy. Variables were incorporated into logistic regression models to predict the pregnancy outcomes, which were evaluated using receiver operator characteristic curves. RESULTS: One hundred and forty-one women were classified as PULs: 67 failing PULs (47.5%), 58 IUPs (41.1%) and 16 ectopic pregnancies (11.4%). Activin A levels were not significantly different between the three outcome groups. Inhibin A levels were significantly lower in failing PULs. The logistic regression model based on serum inhibin levels gave an area under the curve (AUC) of 0.88 for failing PUL, 0.87 for IUP and 0.60 for ectopic pregnancy. The model based on serum activin levels gave an AUC of 0.61 for failing PUL, 0.64 for IUP and 0.51 for ectopic pregnancy, and the model based on serum hCG levels gave an AUC of 0.95 for failing PUL, 0.97 for IUP and 0.67 for ectopic pregnancy. CONCLUSIONS: Serum activin A levels at 0 and 48 h are not helpful in predicting the outcome of PULs. Although serum inhibin A levels may be of use in the prediction of failing PULs and IUPs in the PUL populations, they do not perform as well as serum hCG levels. PMID- 19549701 TI - Large response to cardiac resynchronization therapy in a patient with segmental paradoxical systolic expansion identified by strain imaging. AB - An 84-year-old man with non-ischaemic cardiomyopathy underwent cardiac resynchronization therapy (CRT) based on the presence of drug-refractory heart failure, depressed left ventricular ejection fraction (25%), and wide QRS complex (160 ms). Longitudinal tissue velocity revealed no significant dyssynchrony (23 ms in Yu index and 35 ms in opposing wall delay). However, longitudinal tissue Doppler strain revealed unique appearances in apical four-chamber and long-axis views. The anterior and inferior septum at basal and mid-levels had reversed strain (becoming positive), indicating paradoxical systolic expansion. Ejection fraction dramatically improved from 26 to 50% the day following CRT, and this beneficial effect of CRT was sustained 12 months following CRT. The presence of the segmental reversed strain might have a potential to predict a large response to CRT in the assessment of longitudinal dyssynchrony. PMID- 19549703 TI - High serum follistatin levels in women with ovarian endometriosis. AB - BACKGROUND: Follistatin is an activin-binding protein produced by several tissues, including endometrium and endometriotic implants. We aimed to quantify follistatin in patients with ovarian endometriosis and investigate its value as a diagnostic marker. METHODS: Women undergoing laparoscopic excision of ovarian endometrioma (n = 52) or other benign ovarian cysts (n = 52) were studied, plus women with non-ovarian endometriosis (n = 11) and healthy controls (n = 27). Serum was collected from all subjects, and peritoneal and cystic fluid from a subset with endometrioma. Follistatin was measured by enzyme-linked immunosorbent assay. The diagnostic accuracy of follistatin to detect endometrioma was evaluated by receiver operating characteristic (ROC) curve and compared with cancer antigen (CA)-125. RESULTS: Serum follistatin was increased in women with ovarian endometrioma (2080 +/- 94 pg/ml) compared with controls (545 +/- 49 pg/ml, P < 0.001), other benign ovarian cysts (795 +/- 60 pg/ml, P < 0.001) or non-ovarian endometriosis (1271 +/- 115 pg/ml, P < 0.001). Cystic fluid showed a higher concentration of follistatin (9850 +/- 4461 pg/ml) than peritoneal fluid (1885 +/- 261 pg/ml, P < 0.001) and serum (P < 0.001). Follistatin levels detected 48/52 cases of endometrioma (92% sensitivity) at 1433 pg/ml cut-off, corresponding to 92% specificity. CA-125 detected only 44% of endometriomas with 90% specificity. ROC curve comparison showed follistatin was more accurate than CA-125 to discriminate women with endometrioma either from controls or women with other benign ovarian cysts (P < 0.0001). CONCLUSIONS: Serum follistatin is increased in women with endometriosis and allows clear distinction between endometrioma and other benign ovarian cysts. Follistatin has the sensitivity and specificity to become a useful clinical marker of ovarian endometrioma. PMID- 19549705 TI - Involvement of AdipoR receptor in adiponectin-induced motility and alpha2beta1 integrin upregulation in human chondrosarcoma cells. AB - Chondrosarcoma is a type of highly malignant tumor with a capacity to invade locally and cause distant metastasis. Chondrosarcoma shows a predilection for metastasis to the lungs. Adiponectin is a protein hormone secreted predominantly by differentiated adipocytes and is involved in energy homeostasis. However, the effect of adiponectin on migration activity in human chondrosarcoma cells is mostly unknown. We found that adiponectin increased the migration and expression of alpha2beta1 integrin in human chondrosarcoma cells. The protein and messenger RNA expression of adiponectin receptor (AdipoR1 and AdipoR2) in chondrosarcoma patients and chondrosarcoma cell lines were significantly higher than the normal cartilage. Moreover, primary chondrosarcoma and chondrosarcoma cell lines (SW1353 and JJ012) were more invasive than normal chondrocytes. Adiponectin-mediated migration and integrin expression was attenuated by 5'-adenosine monophosphate activated protein kinase (AMPK) small interfering RNA and an AMPK inhibitor (Ara A and compound C). Activation of p38 and nuclear factor-kappa B (NF-kappaB) pathways after adiponectin treatment was demonstrated, and adiponectin-induced expression of integrins and migration activity was inhibited by the specific inhibitor and mutant of p38 and NF-kappaB cascades. This study showed for the first time that adiponectin mediates the migration of human chondrosarcoma cells. One mechanism underlying adiponectin-directed migration was transcriptional upregulation of alpha2beta1 integrin and activation of AdipoR receptor, AMPK, p38 and NF-kappaB pathways. PMID- 19549704 TI - Benzyl isothiocyanate-mediated generation of reactive oxygen species causes cell cycle arrest and induces apoptosis via activation of MAPK in human pancreatic cancer cells. AB - In our previous studies, we have shown that benzyl isothiocyanate (BITC) inhibits the growth of human pancreatic cancer cells by inducing apoptosis. In the present study, we demonstrate the activation of all the three (MAPK) family members [extracellular signal-regulated protein kinase (ERK), c-jun N-terminal kinase (JNK) and P38] in response to BITC treatment. Exposure of Capan-2 cells with varying concentrations of BITC for 24 h resulted in the phosphorylation (activation) of ERK at Thr202/Tyr204, JNK at Thr183/Tyr185 and P38 at Thr180/Tyr182, leading to the induction of apoptosis. Similar MAPK activation was also observed in MiaPaCa-2 cells in response to BITC treatment. However, normal human pancreatic ductal epithelial cells did not show the activation of MAPK's and remained unaffected by BITC treatment. To confirm the role of ERK, JNK and P38 in BITC-induced G(2)/M arrest and apoptosis, Capan-2 cells were pre-treated with MAPK-specific inhibitors or MAPK8-short hairpin RNA (shRNA) prior to BITC treatment. Significant protection from BITC-induced G(2)/M arrest was observed in the cells pre-treated with MAPK kinase (MEK-1) but not JNK or P38 inhibitors. On the other hand, BITC-induced apoptosis was almost completely abrogated in the cells pre-treated with MEK-1, JNK or P38 inhibitors. Similarly, MAPK8-shRNA also offered almost complete protection against BITC-induced G(2)/M arrest and apoptosis. Furthermore, we observed that BITC treatment leads to the generation of reactive oxygen species (ROS) in Capan-2 and MiaPaCa-2 cells, which in part was orchestrated by depletion of reduced glutathione (GSH) level. Blocking ROS generation with N-acetyl-L-cysteine (NAC) significantly prevented GSH depletion and activation of ERK and JNK but not P38. Further, NAC or tiron prevented G(2)/M arrest by blocking G(2)/M regulatory proteins and completely protected the cells from BITC-induced apoptosis. Taken together, our results suggest that BITC mediated G(2)/M arrest is mediated through ERK activation, whereas apoptosis is via ERK, JNK and P38. PMID- 19549706 TI - Patient-reported outcomes in a phase III, randomized study of sunitinib versus interferon-{alpha} as first-line systemic therapy for patients with metastatic renal cell carcinoma in a European population. AB - BACKGROUND: The purpose of this study is to evaluate the impact on the health related quality of life (HRQoL) of sunitinib versus interferon-alpha (IFN-alpha) treatment in patients with metastatic renal cell carcinoma (mRCC). PATIENTS AND METHODS: In all, 304 mRCC patients (European cohort) were randomized 1 : 1 to receive sunitinib (50 mg/day for 4 weeks, followed by 2 weeks off) or IFN-alpha (9 million units s.c. injection three times/week). The following questionnaires were completed (days 1 and 28 per cycle): Functional Assessment of Cancer Therapy General (FACT-G), the FACT-Kidney Symptom Index and the EuroQol Group's EQ-5D self-report questionnaire (EQ-5D). Results correspond to an ongoing trial with progression-free survival time as primary end point, and patients were still being followed up. Data were analyzed using repeated measures mixed effects models (MEMs) that allow the inclusion of initial differences and uncompleted repeated measures, with the assumption of data missing at random. Six-cycle results were included. RESULTS: Results consistently showed that patients in sunitinib group experienced statistically significantly milder kidney-related symptoms, better cancer-specific HRQoL and general health status (in social utility scores) during the study period as measured by these patient-reported outcome end points. No statistical differences between groups were found on the FACT-G physical well-being subscale or the EQ-5D VAS values. CONCLUSIONS: Results from MEM showed the sunitinib's benefit on HRQoL compared with IFN-alpha. PMID- 19549707 TI - Cetuximab plus cisplatin-5-fluorouracil versus cisplatin-5-fluorouracil alone in first-line metastatic squamous cell carcinoma of the esophagus: a randomized phase II study of the Arbeitsgemeinschaft Internistische Onkologie. AB - BACKGROUND: This study assessed the activity of the mAb cetuximab in combination with cisplatin and 5-fluorouracil (5-FU) in advanced esophageal squamous cell carcinoma. PATIENTS AND METHODS: For a maximum of six 29-day cycles, patients received cisplatin 100 mg/m(2), day 1, plus 5-FU 1000 mg/m(2), days 1-5 (CF), either alone or in combination with cetuximab (CET-CF; 400 mg/m(2) initial dose followed by 250 mg/m(2) weekly thereafter). The primary end point was tumor response. Tumor material was obtained for analysis of KRAS mutation status. RESULTS: Sixty-two eligible patients were included, 32 receiving CET-CF and 30 CF. Cetuximab did not exacerbate grade 3/4 toxicity, except for rash (6% versus 0%) and diarrhea (16% versus 0%). The overall response rate according to RECIST criteria was 19% and 13% and the disease control rate 75% and 57% for the CET-CF and CF arms, respectively. With a median follow-up of 21.5 months, the median progression-free survival was 5.9 and 3.6 months and median overall survival 9.5 and 5.5 months for CET-CF and CF, respectively. No KRAS codon 12/13 tumor mutations were identified in 37 evaluated samples. CONCLUSION: Cetuximab can be safely combined with CF chemotherapy and may increase the efficacy of standard CF chemotherapy. PMID- 19549708 TI - Empirically derived psychosocial states among adolescents diagnosed with cancer during the acute and extended phase of survival. AB - BACKGROUND: To, during the acute and extended phase of survival, identify psychosocial states among adolescents diagnosed with cancer and to analyse these in relation to demographic and clinical characteristics and self-reported depression. PATIENTS AND METHODS: Participants completed the Hospital Anxiety and Depression Scale and two subscales, Vitality and Mental Health, in the SF-36 4-8 weeks (T1) (n = 61), 6 (T2) (n = 57), 12 (T3) (n = 50), and 18 (T4) months (n = 48) after diagnosis. I-State as Object of Analysis was used to identify a finite set of states based on three dimensions. Cluster analysis was carried out using Ward's method. RESULTS: Five states were obtained: psychosocial dysfunction (state A) and poor (B), incomplete (C), good (D), and excellent (E) psychosocial function. At T1, more adolescents than expected by chance were in states A (P < 0.05) and C (P < 0.01) and fewer in states D (P < 0.05) and E (P < 0.001). At T4, more adolescents than expected by chance were in state E (P < 0.001) and fewer in state C (P < 0.05). Female gender and being in late adolescence when diagnosed is related to worse psychosocial function. CONCLUSION: The findings provide support for subgroups of adolescents whose level of vitality, mental health, and anxiety differ during the acute and extended phase of survival of cancer. Clinical interventions tailored to the level of impairment as determined by the clusters may result in better psychosocial outcomes. PMID- 19549709 TI - Efficacy of everolimus (RAD001) in patients with advanced NSCLC previously treated with chemotherapy alone or with chemotherapy and EGFR inhibitors. AB - BACKGROUND: Treatment options are scarce in pretreated advanced non-small-cell lung cancer (NSCLC) patients. RAD001, an oral inhibitor of the mammalian target of rapamycin (mTOR), has shown phase I efficacy in NSCLC. METHODS: Stage IIIb or IV NSCLC patients, with two or fewer prior chemotherapy regimens, one platinum based (stratum 1) or both chemotherapy and epidermal growth factor receptor tyrosine kinase inhibitors (stratum 2), received RAD001 10 mg/day until progression or unacceptable toxicity. Primary objective was overall response rate (ORR). Analyses of markers associated with the mTOR pathway were carried out on archival tumor from a subgroup using immunohistochemistry (IHC) and direct mutation sequencing. RESULTS: Eighty-five patients were enrolled, 42 in stratum 1 and 43 in stratum. ORR was 4.7% (7.1% stratum 1; 2.3% stratum 2). Overall disease control rate was 47.1%. Median progression-free survivals (PFSs) were 2.6 (stratum 1) and 2.7 months (stratum 2). Common > or =grade 3 events were fatigue, dyspnea, stomatitis, anemia, and thrombocytopenia. Pneumonitis, probably or possibly related, mainly grade 1/2, occurred in 25%. Cox regression analysis of IHC scores found that only phospho AKT (pAKT) was a significant independent predictor of worse PFS. CONCLUSIONS: RAD001 10 mg/day was well tolerated, showing modest clinical activity in pretreated NSCLC. Evaluation of RAD001 plus standard therapy for metastatic NSCLC continues. PMID- 19549710 TI - Dietary glycemic load and hepatocellular carcinoma with or without chronic hepatitis infection. AB - BACKGROUND: Hepatitis B virus (HBV) and hepatitis C virus (HCV) are the major risk factors for hepatocellular carcinoma (HCC). The association of diabetes mellitus with HCC suggests that dietary glycemic load (GL) may influence HCC risk. We have examined the association between dietary GL and HCC. PATIENTS AND METHODS: We conducted a hospital-based case-control study in Italy in 1999-2002, including 185 HCC cases and 412 controls who answered a validated food frequency questionnaire and provided blood samples. Odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were computed using unconditional multiple logistic regression. RESULTS: We observed a positive association between GL and HCC overall, with an OR of 3.02 (95% CI 1.49-6.12) for the highest quintile of GL compared with the lowest and a significant trend. The OR among HCC cases with evidence of chronic infection with HBV and/or HCV was 3.25 (95% CI 1.46-7.22), while the OR among those with no evidence of infection was 2.45 (95% CI 0.69 8.64), with no significant trend. The association was not explained by the presence of cirrhosis or diabetes. CONCLUSIONS: High dietary GL is associated with increased risk for HCC. The positive association was most pronounced among HCC cases with HBV and/or HCV markers. PMID- 19549711 TI - Significantly higher levels of vascular endothelial growth factor (VEGF) and shorter survival times for patients with primary operable triple-negative breast cancer. AB - BACKGROUND: Triple-negative breast cancer (TNBC) lacking expression of steroid receptors and human epidermal growth factor receptor 2, having chemotherapy as the only therapeutic option, is characterised by early relapses and poor outcome. We investigated intratumoural (i.t.) levels of the pro-angiogenic cytokine vascular endothelial growth factor (VEGF) and survival in patients with TNBC compared with non-TNBC. PATIENTS AND METHODS: VEGF levels were determined by an enzyme immunosorbent assay in a retrospective series consisting of 679 consecutive primary breast cancer patients. RESULTS: Eighty-seven patients (13%) were classified as TNBC and had significantly higher VEGF levels; median value in TNBC was 8.2 pg/microg DNA compared with 2.7 pg/microg DNA in non-TNBC (P < 0.001). Patients with TNBC had statistically significant shorter recurrence-free survival [hazard ratio (HR) = 1.8; P = 0.0023], breast cancer-corrected survival (HR = 2.2; P = 0.004) and overall survival (HR = 1.8; P = 0.005) compared with non-TNBC. Patients with TNBC relapsed earlier than non-TNBC; mean time from diagnosis to first relapse was 18.8 and 30.7 months, respectively. The time between first relapse and death was also shorter in TNBC: 7.5 months versus 17.5 months in non-TNBC (P = 0.087). CONCLUSIONS: Our results show that TNBC have higher i.t. VEGF levels compared with non-TNBC. Ongoing clinical trials will answer if therapy directed towards angiogenesis may be an alternative way to improve outcome in this poor prognosis group. PMID- 19549712 TI - Validation, revision and extension of the Follicular Lymphoma International Prognostic Index (FLIPI) in a population-based setting. AB - BACKGROUND: The aim of this study was to validate the Follicular Lymphoma International Prognostic Index (FLIPI) in a population-based cohort and to study the relevance of revision and extension of the FLIPI. PATIENTS AND METHODS: Data of 353 unselected patients, 1993-2002, in the Eindhoven Cancer Registry, were collected. Follow-up was completed up to 1 January 2006. Multiple imputations for missing covariates were used. Validity was assessed by comparing observed to predicted survival of the original model and of a revised model with other prognostic variables. RESULTS: The original FLIPI stratified our cohort into three different risk groups based on stage, Hb, lactate dehydrogenase, nodal involvement and age. The discrimination between risk groups was not as good as in the original cohort. A model including age in three categories (< or =60/61 70/>70 years) and presence of cardiovascular disease (CVD) (yes/no) resulted in a better prognostic index. The 5-year overall survival rates were 79%, 59% and 28% in the low-, intermediate- and high-risk groups for the extended FLIPI compared with 81%, 66% and 47% for the original FLIPI, respectively. CONCLUSIONS: The performance of the FLIPI was validated in a population-based setting, but could significantly be improved by a more refined coding of age and by including the presence of CVD. PMID- 19549713 TI - Multicenter phase II study of gemcitabine and oxaliplatin in advanced nasopharyngeal carcinoma--correlation with excision repair cross-complementing-1 polymorphisms. AB - BACKGROUND: Nasopharyngeal carcinoma (NPC) is a platinum-sensitive cancer and excision repair cross-complementing group 1 (ERCC1) polymorphisms have been shown to predict survival in several cancers following platinum therapy. PATIENTS AND METHODS: This multicenter study evaluated the activity of oxaliplatin and prolonged infusion of gemcitabine ('GEMOX' regimen) in recurrent NPC. Baseline blood samples were genotyped for the presence of ERCC1-118 gene polymorphisms. RESULTS: Forty-two patients were recruited, of whom most (61%) had metastatic disease. Of the 40 patients evaluated for response, the respective overall response and disease control rates were 56.1% and 90.2%. At a median follow-up of 14.8 months, the respective median overall survival and time to progression were 19.6 months [95% confidence interval (CI) = 12.8-22 months] and 9 months (95% CI = 7.3-10 months). Grade 3-4 toxic effects were uncommon. The distribution of ERCC1-118 genotypes from 29 patients was C/C (n = 17, 40.5%), C/T (n = 10, 23.8%) and T/T (n = 2, 4.8%). No differences in survival or response rates were found between genotypes. CONCLUSIONS: GEMOX is active in the treatment of recurrent NPC. Detection of single-nucleotide gene polymorphisms from genomic DNA in peripheral blood is feasible in NPC and further studies are warranted. PMID- 19549715 TI - Examining resilience of quality of life in the face of health-related and psychosocial adversity at older ages: what is "right" about the way we age? AB - PURPOSE: This article examines resilience at older ages, focusing on the relationships between quality of life (qol) and adversity. Our objectives are to identify (a) the basis of adversity, (b) the characteristics of resilient individuals, and (c) the attributes that attenuate the full impact of adversity. DESIGN AND METHODS: Resilience is defined as flourishing despite adversity. Analysis is carried out in a subsample of the Boyd Orr cohort (aged between 68 and 82 years) using questionnaire data. Adversity was identified as circumstances that produce a significant average decrease in qol (CASP-19 scores). Participants were classified into resilient and vulnerable groups based on high or low qol (CASP-19 scores dichotomized at the median) in the face of significant adversity. Shared characteristics that define these outcomes are reported. Attributes that attenuate the negative impact of adversity were analyzed using stratified logistic regression. RESULTS: Adversity was typified by functional limitation; life getting worse in the domains of health, stress, and general living circumstances; and experiencing a negative life event. The resilient tended to report fewer multiple adversities. Indicators of protective attributes, which also characterized resilient outcomes relative to qol, included good quality relationships (5.105, confidence interval [CI] 95% 1.323-19.699), integration in the community (10.800, 95% CI 1.227-95.014), developmental coping (3.397, 95% CI 1.079-10.690), and adaptive coping styles (3.211, 95% CI 1.041-9.910). IMPLICATION: Overall results indicate that policies that offer access to protection and help minimize adversity exposure where possible will promote resilience. PMID- 19549714 TI - Insulin granule recruitment and exocytosis is dependent on p110gamma in insulinoma and human beta-cells. AB - OBJECTIVE: Phosphatidylinositol 3-OH kinase (PI3K) has a long-recognized role in beta-cell mass regulation and gene transcription and is implicated in the modulation of insulin secretion. The role of nontyrosine kinase receptor activated PI3K isoforms is largely unexplored. We therefore investigated the role of the G-protein-coupled PI3Kgamma and its catalytic subunit p110gamma in the regulation of insulin granule recruitment and exocytosis. RESEARCH DESIGN AND METHODS: The expression of p110gamma was knocked down by small-interfering RNA, and p110gamma activity was selectively inhibited with AS605240 (40 nmol/l). Exocytosis and granule recruitment was monitored by islet perifusion, whole-cell capacitance, total internal reflection fluorescence microscopy, and electron microscopy in INS-1 and human beta-cells. Cortical F-actin was examined in INS-1 cells and human islets and in mouse beta-cells lacking the phosphatase and tensin homolog (PTEN). RESULTS: Knockdown or inhibition of p110gamma markedly blunted depolarization-induced insulin secretion and exocytosis and ablated the exocytotic response to direct Ca(2+) infusion. This resulted from reduced granule localization to the plasma membrane and was associated with increased cortical F actin. Inhibition of p110gamma had no effect on F-actin in beta-cells lacking PTEN. Finally, the effect of p110gamma inhibition on granule localization and exocytosis could be rapidly reversed by agents that promote actin depolymerization. CONCLUSIONS: The G-protein-coupled PI3Kgamma is an important determinant of secretory granule trafficking to the plasma membrane, at least in part through the negative regulation of cortical F-actin. Thus, p110gamma activity plays an important role in maintaining a membrane-docked, readily releasable pool of secretory granules in insulinoma and human beta-cells. PMID- 19549716 TI - Concordance of family and staff member reports about end of life in assisted living and nursing homes. AB - PURPOSE: To identify differences in perspectives that may complicate the process of joint decision making at the end of life, this study determined the agreement of family and staff perspectives about end-of-life experiences in nursing homes and residential care/assisted living communities and whether family and staff roles, involvement in care, and interaction are associated with such agreement. DESIGN AND METHODS: This cross-sectional study examined agreement in 336 family staff pairs of postdeath telephone interviews conducted as part of the Collaborative Studies of Long-Term Care. Eligible deaths occurred in or within 3 days of leaving one of a stratified random sample of 113 long-term care facilities in four states and after the resident had lived in the facility (3)15 days of the last month of life. McNemar p values and kappas were determined for each concordance variable, and mixed logistic models were run. RESULTS: Chance adjusted family-staff agreement was poor for expectation of death within weeks (66.9% agreement, kappa = .33), course of illness (62.9%, 0.18), symptom burden (59.6%, 0.18), and familiarity with resident's physician (59.2%, 0.05). Staff were more likely than family to expect death (70.2% vs 51.5%, p < .001) and less likely to report low symptom burden (39.6% vs 46.6%, p = .07). Staff involvement in care related to concordance and perspectives of adult children were more similar to those of staff than were other types of family members. IMPLICATIONS: Family and staff perspectives about end-of-life experiences may differ substantially; efforts can be made to improve family-staff communication and interaction for joint decision making. PMID- 19549717 TI - Differential modification of Cys10 alters transthyretin's effect on beta-amyloid aggregation and toxicity. AB - Tg2576 mice produce high levels of beta-amyloid (Abeta) and develop amyloid deposits, but lack neurofibrillary tangles and do not suffer the extensive neuronal cell loss characteristic of Alzheimer's disease. Protection from Abeta toxicity has been attributed to up-regulation of transthyretin (TTR), a normal component of plasma and cerebrospinal fluid. We compared the effect of TTR purified from human plasma (pTTR) with that produced recombinantly (rTTR) on Abeta aggregation and toxicity. pTTR slowed Abeta aggregation but failed to protect primary cortical neurons from Abeta toxicity. In contrast, rTTR accelerated aggregation, while effectively protecting neurons. This inverse correlation between Abeta aggregation kinetics and toxicity is consistent with the hypothesis that soluble intermediates rather than insoluble fibrils are the most toxic Abeta species. We carried out a detailed comparison of pTTR with rTTR to ascertain the probable cause of these different effects. No differences in secondary, tertiary or quaternary structure were detected. However, pTTR differed from rTTR in the extent and nature of modification at Cys10. We hypothesize that differential modification at Cys10 regulates TTR's effect on Abeta aggregation and toxicity. PMID- 19549718 TI - Breaking the seals: efficient mRNA detection from human archival paraffin embedded tissue. AB - During our study on HOXA13, HOXD12, and HOXD13 mRNA expression in human adult and embryonic tissues, we were confronted with the fact that, within our specimen collection, as in other University Departments in Europe, <20% of all samples yielded reliable labeling, while most samples were resistant to hybridization by standard protocols due to over-fixation. Fixation is essential for specimen stability, especially when samples are stored at room temperature and used for histology, and people tend to be more worried about under- than over-fixation. On the other hand fixation inhibits penetration by the probe and may also trap mRNA within ribosomes. Therefore, we developed a nonradioactive in situ hybridization technique, which allows detection of mRNA expressed on low levels from a variety of differentially fixed tissues while maintaining tissue integrity. This was achieved by improving target retrieval and probe detection. In contrast with others, our method allows reliable staining from tissues that are fixed in paraformaldehyde from four hours to over one week, and archived samples that were stored at room temperature for several years (17-19 yr in some cases) and exceeds detection limits of purely fluorescent methods. Our protocol is highly suitable for detecting CDX-2 mRNA in carcinoma specimens, but especially designed to investigate mRNAs in nonpathological adult and embryonic tissues. Due to the use of standardized probes, we do not expect problems in detecting other mRNAs expressed in suitable amounts. PMID- 19549720 TI - A phase II trial of chemoradiotherapy for stage I esophageal squamous cell carcinoma: Japan Clinical Oncology Group Study (JCOG9708). AB - OBJECTIVE: The study objective was to evaluate the efficacy and toxicity of chemoradiotherapy with 5-fluorouracil (5-FU) plus cisplatin in patients with Stage I esophageal squamous cell carcinoma (ESCC). The primary endpoint was proportion of complete response (%CR). METHODS: Patients with Stage I (T1N0M0) ESCC, aged 20-75 years, without indication of endoscopic mucosal resection were eligible. Treatment consisted of cisplatin 70 mg/m(2) (day 1) and 5-FU 700 mg/m(2)/day (days 1-4) combined with 30 Gy radiotherapy (2 Gy/day, 5 days/week, days 1-21). The cycle was repeated twice with 1-week split. Salvage surgery was recommended for residual tumor or local recurrence. RESULTS: From December 1997 to June 2000, 72 patients were enrolled. No ineligible patient or major protocol violation was observed. There were 63 CRs for %CR of 87.5% [95% confidence interval (CI): 77.6-94.1]. Six patients with residual tumor successfully underwent esophagectomy. There was no Grade 4 toxicity. Four-year survival proportion was 80.5% (95% CI: 71.3-89.7), and 4-year major relapse-free survival proportion was 68% (95% CI: 57.3-78.8) (mucosal recurrence removed by endoscopy was not counted as an event). CONCLUSIONS: High CR proportion and survival proportion with mild toxicity suggest that this regimen could be considered as a candidate of new standard treatment to be compared with surgery in patients with Stage I ESCC. PMID- 19549722 TI - Repeatable battery for the assessment of neuropsychological status effort index performance in a medically ill geriatric sample. AB - Using a sample of 44 clinically referred, non-litigating, older adults, we evaluated the Repeatable Battery for the Assessment of Neuropsychological Status Effort Index [RBANS EI; Silverberg, N. D., Wertheimer, J. C., & Fichtenberg, N. L. (2007). An effort index for the Repeatable Battery for the Assessment of Neurospcyhological Status (RBANS). The Clinical Neuropsychologist, 21, 841-854]. With the current RBANS EI cut-score guidelines, 31% of our sample was classified as putting forth suspect effort. With this, cognitive ability was significantly correlated with suspect effort scores. Thus, it appears that the current guidelines may not be useful in a cognitively impaired medically ill geriatric sample. Hence, further research on RBANS EI validation is warranted. PMID- 19549719 TI - Conformational change in the Bacillus subtilis RNase P holoenzyme--pre-tRNA complex enhances substrate affinity and limits cleavage rate. AB - Ribonuclease P (RNase P) is a ribonucleoprotein complex that catalyzes the 5' maturation of precursor tRNAs. To investigate the mechanism of substrate recognition in this enzyme, we characterize the thermodynamics and kinetics of Bacillus subtilis pre-tRNA(Asp) binding to B. subtilis RNase P holoenzyme using fluorescence techniques. Time courses for fluorescein-labeled pre-tRNA binding to RNase P are biphasic in the presence of both Ca(II) and Mg(II), requiring a minimal two-step association mechanism. In the first step, the apparent bimolecular rate constant for pre-tRNA associating with RNase P has a value that is near the diffusion limit and is independent of the length of the pre-tRNA leader. Following formation of the initial enzyme-substrate complex, a unimolecular step enhances the overall affinity of pre-tRNA by eight- to 300-fold as the length of the leader sequence increases from 2 to 5 nucleotides. This increase in affinity is due to a decrease in the reverse rate constant for the conformational change that correlates with the formation of an optimal leader protein interaction in the RNase P holoenzyme-pre-tRNA complex. Furthermore, the forward rate constant for the conformational change becomes rate limiting for cleavage under single-turnover conditions at high pH, explaining the origin of the observed apparent pK(a) in the RNase P-catalyzed cleavage reaction. These data suggest that a conformational change in the RNase P*pre-tRNA complex is coupled to the interactions between the 5' leader and P protein and aligns essential functional groups at the cleavage active site to enhance efficient cleavage of pre-tRNA. PMID- 19549721 TI - Concussion symptom inventory: an empirically derived scale for monitoring resolution of symptoms following sport-related concussion. AB - Self-report post-concussion symptom scales have been a key method for monitoring recovery from sport-related concussion, to assist in medical management, and return-to-play decision-making. To date, however, item selection and scaling metrics for these instruments have been based solely upon clinical judgment, and no one scale has been identified as the "gold standard". We analyzed a large set of data from existing scales obtained from three separate case-control studies in order to derive a sensitive and efficient scale for this application by eliminating items that were found to be insensitive to concussion. Baseline data from symptom checklists including a total of 27 symptom variables were collected from a total of 16,350 high school and college athletes. Follow-up data were obtained from 641 athletes who subsequently incurred a concussion. Symptom checklists were administered at baseline (preseason), immediately post concussion, post-game, and at 1, 3, and 5 days post-injury. Effect-size analyses resulted in the retention of only 12 of the 27 variables. Receiver-operating characteristic analyses were used to confirm that the reduction in items did not reduce sensitivity or specificity. The newly derived Concussion Symptom Inventory is presented and recommended as a research and clinical tool for monitoring recovery from sport-related concussion. PMID- 19549725 TI - Who can resist Foucault? AB - Michel Foucault's analysis of "the birth of the clinic" describes the genesis of a unified discourse that, in retrospect, has shaped western medicine for two centuries. However, in looking prospectively toward a 21st century medicine, Foucault's analysis is necessary but not sufficient. To better critically address medicine and medical education in the era of simulation, we could draw on frameworks developed by futurists such as Jean Baudrillard. Foucault's analysis does not account for contemporary, complex developments of the clinical gaze as the gaze is distributed across practitioners in increasing use of sophisticated, representational diagnostic imaging. Further, Foucault's antihumanist rhetoric sometimes strays into the antihumane, and this is disturbing for those who support the development of patient-centered medicine. Yet we are increasingly teaching aspects of medicine, such as communication, in simulated learning environments in which complex reality is absent, perhaps inadvertently creating an "inhumanity" in medical education. PMID- 19549724 TI - Memory functioning in developmental dyslexia: an analysis using two clinical memory measures. AB - The goals of this project were threefold: to determine the nature of the memory deficit in children/adolescents with dyslexia, to utilize clinical memory measures in this endeavor, and to determine the extent to which semantic short term memory (STM) is related to basic reading performance. Two studies were conducted using different samples, one incorporating the Wide Range Assessment of Memory and Learning and the other incorporating the California Verbal Learning Test-Children's Version. Results suggest that phonological STM is deficient in children with dyslexia, but semantic STM and visual-spatial STM are intact. Long term memory (LTM) for both visual and verbal material also is intact. Regarding reading performance, semantic STM had small correlations with word identification and pseudoword decoding across studies despite phonological STM being moderately to strongly related to both basic reading skills. Overall, results are consistent with the phonological core deficit model of dyslexia as only phonological STM was affected in dyslexia and related to basic reading skill. PMID- 19549723 TI - Spanish Multicenter Normative Studies (NEURONORMA Project): methods and sample characteristics. AB - This paper describes the methods and sample characteristics of a series of Spanish normative studies (The NEURONORMA project). The primary objective of our research was to collect normative and psychometric information on a sample of people aged over 49 years. The normative information was based on a series of selected, but commonly used, neuropsychological tests covering attention, language, visuo-perceptual abilities, constructional tasks, memory, and executive functions. A sample of 356 community dwelling individuals was studied. Demographics, socio-cultural, and medical data were collected. Cognitive normality was validated via informants and a cognitive screening test. Norms were calculated for midpoint age groups. Effects of age, education, and sex were determined. The use of these norms should improve neuropsychological diagnostic accuracy in older Spanish subjects. These data may also be of considerable use for comparisons with other normative studies. Limitations of these normative data are also commented on. PMID- 19549726 TI - Docile bodies: transnational research ethics as biopolitics. AB - This essay explores the claim that bioethics has become a mode of biopolitics. It seeks to illuminate one of the myriad of ways that bioethics joins other institutionalized discursive practices in the task of producing, organizing, and managing the bodies-of policing and controlling populations-in order to empower larger institutional agents. The focus of this analysis is the contemporary practice of transnational biomedical research. The analysis is catalyzed by the enormous transformation in the political economy of transnational research that has occurred over the past three decades and the accompanying increase in the numbers of human bodies now subjected to research. This essay uses the work of Michel Foucault, particularly his notion of docile bodies, to analyze these changes. Two loci from the bioethics literature are explored-one treating research in the United States and one treating research in developing countries. In the latter, we see a novel dynamic of the new biopolitics: the ways in which bioethics helps to create docile political bodies that will police themselves and who will, in turn, facilitate the production of docile human bodies for research. PMID- 19549727 TI - Analysis of the human E2 ubiquitin conjugating enzyme protein interaction network. AB - In eukaryotic cells the stability and function of many proteins are regulated by the addition of ubiquitin or ubiquitin-like peptides. This process is dependent upon the sequential action of an E1-activating enzyme, an E2-conjugating enzyme, and an E3 ligase. Different combinations of these proteins confer substrate specificity and the form of protein modification. However, combinatorial preferences within ubiquitination networks remain unclear. In this study, yeast two-hybrid (Y2H) screens were combined with true homology modeling methods to generate a high-density map of human E2/E3-RING interactions. These data include 535 experimentally defined novel E2/E3-RING interactions and >1300 E2/E3-RING pairs with more favorable predicted free-energy values than the canonical UBE2L3 CBL complex. The significance of Y2H predictions was assessed by both mutagenesis and functional assays. Significantly, 74/80 (>92%) of Y2H predicted complexes were disrupted by point mutations that inhibit verified E2/E3-RING interactions, and a approximately 93% correlation was observed between Y2H data and the functional activity of E2/E3-RING complexes in vitro. Analysis of the high density human E2/E3-RING network reveals complex combinatorial interactions and a strong potential for functional redundancy, especially within E2 families that have undergone evolutionary expansion. Finally, a one-step extended human E2/E3 RING network, containing 2644 proteins and 5087 edges, was assembled to provide a resource for future functional investigations. PMID- 19549728 TI - First-trimester fasting hyperglycemia and adverse pregnancy outcomes. AB - OBJECTIVE: The Hyperglycemia and Adverse Pregnancy Outcome (HAPO) study found strong associations between higher levels of maternal glucose at 24-32 weeks, within what is currently considered normoglycemia and adverse pregnancy outcomes. Our aim was to evaluate the associations between first-trimester fasting plasma glucose level and adverse pregnancy outcomes. RESEARCH DESIGN AND METHODS: Charts of all patients who delivered at our hospital between June 2001 and June 2006 were reviewed. Only subjects with singleton pregnancy and a recorded first trimester fasting glucose level were included. Women with pregestational diabetes, fasting glucose level >105 mg/dl, or delivery <24 weeks were excluded. Fasting glucose levels were analyzed in seven categories, similar to the HAPO study. The main outcomes were development of gestational diabetes mellitus (GDM), large-for-gestational-age (LGA) neonates and/or macrosomia, and primary cesarean section. Multivariate logistic regression analysis was used; significance was <0.05. RESULTS: A total of 6,129 women had a fasting glucose test at median of 9.5 weeks. There were strong, graded associations between fasting glucose level and primary outcomes. The frequency of GDM development increased from 1.0% in the lowest glucose category to 11.7% in the highest (adjusted odds ratio 11.92 [95% CI 5.39-26.37]). The frequency of LGA neonates and/or macrosomia increased from 7.9 to 19.4% (2.82 [1.67-4.76]). Primary cesarean section rate increased from 12.7 to 20.0% (1.94 [1.11-3.41]). CONCLUSIONS: Higher first-trimester fasting glucose levels, within what is currently considered a nondiabetic range, increase the risk of adverse pregnancy outcomes. Early detection and treatment of women at high risk for these complications might improve pregnancy outcome. PMID- 19549730 TI - Ketoacidosis at diabetes onset is still frequent in children and adolescents: a multicenter analysis of 14,664 patients from 106 institutions. AB - OBJECTIVE: We aimed at analyzing the frequency, clinical characteristics, and trends associated with the occurrence of diabetic ketoacidosis (DKA) at the onset of type 1 diabetes on the basis of long-term follow-up data. RESEARCH DESIGN AND METHODS: A total of 106 pediatric diabetes centers in Germany and Austria participated in this study. Data from 14,664 patients with type 1 diabetes collected between 1995 and 2007 were suitable for evaluation. DKA was defined and classified according to the International Society for Pediatric and Adolescent Diabetes consensus guidelines. RESULTS: DKA was observed in 21.1% of patients. The frequency of DKA, including the severe form, remained unchanged throughout the 13-year observation period. The frequency of DKA was particularly striking among children <5 years of age (26.5%). CONCLUSIONS: Ketoacidosis occurring at diabetes onset continues to be a difficult problem. Our data show no significant change in the frequency and magnitude of DKA over the last 13 years. PMID- 19549729 TI - Association between serum uric acid and development of type 2 diabetes. AB - OBJECTIVE: To systematically evaluate the association between serum uric acid (SUA) level and subsequent development of type 2 diabetes. RESEARCH DESIGN AND METHODS: We searched Medline (31 March from 1966 to 2009) and Embase (31 March from 1980 to 2009) for observational cohort studies examining the association between SUA and the risk of type 2 diabetes by manual literature search. Relative risks (RRs) for each 1 mg/dl increase in SUA were pooled by using a random effects model. The studies included were stratified into subgroups representing different study characteristics, and meta-regression analyses were performed to investigate the effect of these characteristics on the association between SUA level and type 2 diabetes risk. RESULTS: The search yielded 11 cohort studies (42,834 participants) that reported 3,305 incident cases of type 2 diabetes during follow-up periods ranging from 2.0 to 13.5 years. The pooled RR of a 1 mg/dl increase in SUA was 1.17 (95% CI 1.09-1.25). Study results were consistently significant (i.e., >1) across characteristics of participants and study design. Publication bias was both visually and statistically suggested (P = 0.03 for Egger's test, 0.06). Adjustment for publication bias attenuated the pooled RR per mg/dl increase in SUA (RR 1.11 [95% CI 1.03-1.20]), but the association remained statistically significant (P = 0.009). CONCLUSIONS: The current meta-analysis suggests that SUA level is positively associated with the development of type 2 diabetes regardless of various study characteristics. Further research should attempt to determine whether it is effective to utilize SUA level as a predictor of type 2 diabetes for its primary prevention. PMID- 19549731 TI - Safety and viability of microencapsulated human islets transplanted into diabetic humans. AB - OBJECTIVE: Transplantation of insulin-producing cells placed inside microcapsules is being trialled to overcome the need for immunosuppressive therapy. RESEARCH DESIGN AND METHODS: Four type 1 diabetic patients with no detectable C-peptide received an intraperitoneal infusion of islets inside microcapsules of barium alginate (mean 178,200 islet equivalents on each of eight occasions). RESULTS: C peptide was detected on day 1 post-transplantation, and blood glucose levels and insulin requirements decreased. C-peptide was undetectable by 1-4 weeks. In a multi-islet recipient, C-peptide was detected at 6 weeks after the third infusion and remains detectable at 2.5 years. Neither insulin requirements nor glycemic control was affected. Capsules recovered at 16 months were surrounded by fibrous tissue and contained necrotic islets. No major side effects or infection occurred. CONCLUSIONS: While allografting of encapsulated human islets is safe, efficacy of the cells needs to improve for the therapy to make an impact on the clinical scene. PMID- 19549732 TI - The association between A1C and subclinical cardiovascular disease: the multi ethnic study of atherosclerosis. AB - OBJECTIVE: To test the hypothesis that A1C is associated with subclinical cardiovascular disease (CVD) in a population without evident diabetes, after adjusting for traditional CVD risk factors and BMI. RESEARCH DESIGN AND METHODS: This was a cross-sectional study of 5,121 participants without clinically evident CVD or diabetes (fasting glucose > or =7.0 mmol/l or use of diabetes medication), aged 47-86 years, enrolled in the Multi-Ethnic Study of Atherosclerosis (MESA). Measurements included carotid intimal-medial wall thickness (CIMT) and coronary artery calcification (CAC). Results were adjusted for age, sex, ethnicity, smoking, systolic blood pressure, LDL cholesterol, HDL cholesterol, antihypertensive medication use, lipid-lowering medication use, and BMI. RESULTS: Compared with those in the lowest quartile for A1C ([mean +/- SD] 5.0 +/- 0.2%), participants in the highest quartile (6.0 +/- 0.3%) had higher adjusted mean values for common CIMT (0.85 vs. 0.87 mm, P = 0.003) and internal CIMT (1.01 vs. 1.08 mm, P = 0.003). A1C quartile was not associated with prevalence of CAC in the entire cohort (P = 0.27); however, the association was statistically significant in women (adjusted prevalence of CAC in lowest and highest A1C quartiles 37.5 vs. 43.0%, P = 0.01). Among those with some CAC, higher A1C quartile tended to be associated with higher CAC score, but the results were not statistically significant (adjusted P = 0.11). CONCLUSIONS: In this multiethnic cohort, there were small, positive associations between A1C, common CIMT, and internal CIMT in the absence of clinically evident diabetes. An association between higher A1C and CAC prevalence was evident only in women. PMID- 19549733 TI - Inflammatory, hemostatic, and other novel biomarkers for diabetic retinopathy: the multi-ethnic study of atherosclerosis. AB - OBJECTIVE: There are conflicting data regarding relationships of systemic biomarkers of inflammation, hemostasis, and homocysteine with diabetic retinopathy. We examined these relationships in the Multi-Ethnic Study of Atherosclerosis. RESEARCH DESIGN AND METHODS: A total of 921 participants with diabetes were included. Diabetic retinopathy was graded from retinal photographs. We defined two outcomes: any diabetic retinopathy and vision-threatening diabetic retinopathy (severe nonproliferative diabetic retinopathy or worse). Systemic markers analyzed were C-reactive protein, homocysteine, fibrinogen, plasmin alpha(2)-antiplasmin complex (PAP), interleukin-6, d-dimer, factor VIII, serum creatinine, and urinary albumin-to-creatinine (UAC) ratio. RESULTS: Prevalence of diabetic retinopathy was 33.2% and vision-threatening diabetic retinopathy 7.1%. After adjusting for established risk factors (diabetes duration, A1C, systolic blood pressure, waist-to-hip ratio, and use of diabetes medications), fibrinogen (odds ratio 1.14 [95% CI 1.01-1.32], P = 0.05) and PAP (1.25 [1.05-1.50], P = 0.01) were associated with any diabetic retinopathy, while PAP (1.54 [1.13-2.11], P = 0.007) and homocysteine (1.57 [1.16-2.11], P = 0.003) were associated with vision-threatening diabetic retinopathy. Only PAP remained significant after additional adjustment for serum creatinine and UAC ratio. Area under receiver operator characteristic curve (AUROC) for diabetic retinopathy was constructed for established and novel risk factors. Established risk factors accounted for a 39.2% increase of the AUROC, whereas novel markers (fibrinogen, PAP, homocysteine, serum creatinine, and UAC ratio) only accounted for an additional 2.2%. CONCLUSIONS: There were few associations of novel markers of inflammation, hemostasis, and homocysteine with diabetic retinopathy after controlling for established risk factors. These data suggest that there is limited clinical use of these biomarkers for prediction of diabetic retinopathy. PMID- 19549734 TI - Excessive loss of skeletal muscle mass in older adults with type 2 diabetes. AB - OBJECTIVE: A loss of skeletal muscle mass is frequently observed in older adults. The aim of the study was to investigate the impact of type 2 diabetes on the changes in body composition, with particular interest in the skeletal muscle mass. RESEARCH DESIGN AND METHODS: We examined total body composition with dual energy X-ray absorptiometry annually for 6 years in 2,675 older adults. We also measured mid-thigh muscle cross-sectional area (CSA) with computed tomography in year 1 and year 6. At baseline, 75-g oral glucose challenge tests were performed. Diagnosed diabetes (n = 402, 15.0%) was identified by self-report or use of hypoglycemic agents. Undiagnosed diabetes (n = 226, 8.4%) was defined by fasting plasma glucose (>or=7 mmol/l) or 2-h postchallenge plasma glucose (>or=11.1 mmol/l). Longitudinal regression models were fit to examine the effect of diabetes on the changes in body composition variables. RESULTS: Older adults with either diagnosed or undiagnosed type 2 diabetes showed excessive loss of appendicular lean mass and trunk fat mass compared with nondiabetic subjects. Thigh muscle CSA declined two times faster in older women with diabetes than their nondiabetic counterparts. These findings remained significant after adjusting for age, sex, race, clinic site, baseline BMI, weight change intention, and actual weight changes over time. CONCLUSIONS: Type 2 diabetes is associated with excessive loss of skeletal muscle and trunk fat mass in community-dwelling older adults. Older women with type 2 diabetes are at especially high risk for loss of skeletal muscle mass. PMID- 19549735 TI - Graves hyperthyroidism after stopping immunosuppressive therapy in type 1 diabetic Islet cell recipients with pretransplant TPO autoantibodies. AB - OBJECTIVE: After an initially successful islet cell transplantation, a number of patients return to C-peptide negativity, and therefore immunosuppressive therapy is discontinued. Some are then found to have developed Graves disease. We examined the risk of Graves disease after immunosuppression. RESEARCH DESIGN AND METHODS: Immunosuppressive therapy was stopped in 13 type 1 diabetic islet cell recipients who had received one course of antithymocyte globulin and maintenance doses of mycophenolate mofetil and a calcineurin inhibitor. None had a history of thyroid disease. RESULTS: In four patients, clinical Graves hyperthyroidism was observed within 21 months after discontinuation and 30-71 months after the start of immunosuppressive therapy. All four patients exhibited a pretransplant positivity for thyroid peroxidase (TPO) autoantibodies, while the nine others were TPO negative pre- and posttransplantation. CONCLUSIONS: Type 1 diabetic recipients of islet cell grafts with pretransplant TPO autoantibody positivity exhibit a high risk for developing Graves hyperthyroidism after immunosuppressive therapy is discontinued for a failing graft. PMID- 19549736 TI - Effect of glucose variability on the long-term risk of microvascular complications in type 1 diabetes. AB - OBJECTIVE: This study analyzed data from the Epidemiology of Diabetes Interventions and Complications (EDIC) study to see whether longer-term follow-up of Diabetes Control and Complications Trial (DCCT) patients reveals a role for glycemic instability in the development of microvascular complications. RESEARCH DESIGN AND METHODS: The mean area under the curve glucose and the within-day glucose variability (SD and mean amplitude of glycemic excursions [MAGE]) during the DCCT were assessed to see whether they contributed to the risk of retinopathy and nephropathy by year 4 of the EDIC. RESULTS: Logistic regression analysis showed that mean glucose during the DCCT and mean A1C during EDIC were independently predictive of retinopathy (each P < 0.001) as well as A1C during EDIC of nephropathy (P = 0.001) development by EDIC year 4. Glucose variability did not add to this (all P > 0.25 using SD or MAGE). CONCLUSIONS: Glucose variability in the DCCT did not predict the development of retinopathy or nephropathy by EDIC year 4. PMID- 19549737 TI - Effects of salt supplementation on the albuminuric response to telmisartan with or without hydrochlorothiazide therapy in hypertensive patients with type 2 diabetes are modulated by habitual dietary salt intake. AB - OBJECTIVE This prospective randomized double-blind placebo-controlled crossover study examined the effects of sodium chloride (NaCl) supplementation on the antialbuminuric action of telmisartan with or without hydrochlorothiazide (HCT) in hypertensive patients with type 2 diabetes, increased albumin excretion rate (AER), and habitual low dietary salt intake (LDS; <100 mmol sodium/24 h on two of three consecutive occasions) or high dietary salt intake (HDS; >200 mmol sodium/24 h on two of three consecutive occasions). RESEARCH DESIGN AND METHODS Following a washout period, subjects (n = 32) received 40 mg/day telmisartan for 4 weeks followed by 40 mg telmisartan plus 12.5 mg/day HCT for 4 weeks. For the last 2 weeks of each treatment period, patients received either 100 mmol/day NaCl or placebo capsules. After a second washout, the regimen was repeated with supplements in reverse order. AER and ambulatory blood pressure were measured at weeks 0, 4, 8, 14, 18, and 22. RESULTS In LDS, NaCl supplementation reduced the anti-albuminuric effect of telmisartan with or without HCT from 42.3% (placebo) to 9.5% (P = 0.004). By contrast, in HDS, NaCl supplementation did not reduce the AER response to telmisartan with or without HCT (placebo 30.9%, NaCl 28.1%, P = 0.7). Changes in AER were independent of changes in blood pressure. CONCLUSIONS The AER response to telmisartan with or without HCT under habitual low salt intake can be blunted by NaCl supplementation. By contrast, when there is already a suppressed renin angiotensin aldosterone system under habitual high dietary salt intake, the additional NaCl does not alter the AER response. PMID- 19549739 TI - Transdermal nitroglycerin therapy may not prevent early postmenopausal bone loss. AB - CONTEXT: Osteoporosis is common among postmenopausal women; animal studies and human pilot studies support the concept of nitric oxide (NO) donors reducing bone mineral density loss. OBJECTIVE: The objective of the study was to evaluate whether NO donor, nitroglycerin, prevents postmenopausal bone loss. DESIGN: This was a 3-yr randomized, double blinded, single-center, placebo-controlled clinical trial. SETTING: The single-center study was conducted at the University of Medicine and Dentistry-Robert Wood Johnson Medical School (New Brunswick, NJ). PARTICIPANTS: Participants included 186 postmenopausal women aged 40-65 yr, with lumbar bone mineral density (BMD) T-scores of 0 to -2.5. INTERVENTION: Women, stratified by lumbar T-score (<-1.50 and >or=-1.50) and years since menopause (5 yr), were randomized to receive nitroglycerin ointment (22.5 mg as Nitro-Bid) or placebo ointment received daily for 3 yr. Both groups took 630 mg daily calcium plus 400 IU vitamin D supplements. MEASUREMENTS: BMD was measured at 6 months and annually by dual-energy x-ray absorptiometry. Percent change in lumbar vertebrae BMD was the primary outcome. Hip BMD, total body bone mineral content, and height were secondary outcomes. RESULTS: After 36 months of therapy, changes of -2.1% in the active group (n = 88) and -2.5% in the placebo group (n = 82) in lumbar spine BMD were seen (P = 0.59; 95% confidence interval -1.001, 1.975). Secondary outcomes also did not differ by intervention arm. The active group reported more headaches compared with the placebo group (57 vs. 14%, P < 0.001). Other adverse and serious adverse events were not different. CONCLUSIONS: BMD changes did not substantially differ between postmenopausal women who received the dose of nitroglycerin tested, in comparison with a placebo. Once daily dosing with 22.5 mg of transdermal-administered nitroglycerin was not effective (compliance adjusted dose was only approximately 16 mg/d); a sub therapeutic dose. PMID- 19549738 TI - Association of plasma vitamin D levels with adiposity in Hispanic and African Americans. AB - CONTEXT: Previous studies have suggested vitamin D insufficiency is associated with increased obesity; however, the relationship between 25-hydroxyvitamin D (25[OH]D) and 1,25-dihydroxyvitamin D (1,25[OH](2)D) and measures of adiposity has not been well characterized in minority populations. OBJECTIVE: The objective of the study was to examine the relationship between levels of 25[OH]D and 1,25[OH](2)D and measures of adiposity in Hispanic and African-Americans at baseline and on change in these measures over time. DESIGN AND SETTING: The Insulin Resistance Atherosclerosis (IRAS) Family Study examined 917 Hispanics and 439 African-Americans at baseline and again 5.3 yr later (n = 1081 at follow-up). MAIN OUTCOME MEASURE: 25[OH]D (nanograms per milliliter) and 1,25[OH](2)D (picograms per milliliter) were measured at baseline. Abdominal sc adipose tissue (SAT), visceral adipose tissue (VAT; both determined by computed tomography scan), and body mass index (BMI) were measured at baseline and follow-up. RESULTS: 25[OH]D was inversely associated with BMI, VAT, and SAT in both populations at baseline (P < 0.001). 25[OH]D was marginally inversely associated with baseline visceral fat to sc fat ratio in African-Americans (P = 0.049) but not Hispanics. 1,25[OH](2)D was inversely associated with BMI (P < 0.0001, P = 0.002) and VAT (P = 0.0005, P = 0.012) in Hispanics and African-Americans, respectively, whereas 1,25[OH](2)D was inversely associated with SAT in Hispanics (P < 0.0001) and with visceral fat to sc fat ratio in African-Americans (P = 0.02). Adjusting for 25[OH]D attenuated these associations; 1,25[OH](2)D remained associated with BMI in both populations (P < 0.05) and with SAT (P = 0.004) in Hispanics. No significant associations between 5-yr change in adiposity and 25[OH]D or 1,25[OH](2)D were seen. CONCLUSIONS: Vitamin D levels were inversely associated with baseline BMI, SAT, and VAT in Hispanic and African-Americans but were not associated with 5-yr change in adiposity. PMID- 19549740 TI - Aging attenuates the pituitary response to gonadotropin-releasing hormone. AB - CONTEXT: Complex changes in GnRH secretion occur with aging in women, but little is known about the effect of aging on the pituitary per se. OBJECTIVE: The aim of the study was to determine whether pituitary responsiveness to GnRH is attenuated with aging. DESIGN AND SETTING: A GnRH antagonist and graded doses of GnRH were used to isolate pituitary responsiveness in Clinical Research Center studies at an academic medical center. SUBJECTS: Subjects were healthy postmenopausal women (PMW) aged 48-57 yr (n = 10) or 70-77 yr (n= 9). INTERVENTIONS: A suppressive dose of the NAL-GLU GnRH antagonist (150 microg/kg sc) was administered and was followed by GnRH doses of 25, 75, 250, or 750 ng/kg iv every 4 h. RESULTS: The LH response to GnRH was attenuated with aging (P = 0.05) with an interaction between age and dose (P = 0.01) such that the LH amplitude was less in older PMW at the higher doses (250 ng/kg, 50 +/- 9 vs. 29 +/- 4.9 IU/liter, for young and old PMW, respectively, P = 0.02; and 750 ng/kg, 97.7 +/- 11 vs. 70.2 +/- 9.3 IU/liter, P = 0.002), but not the lower doses of GnRH. The FSH response to GnRH was also attenuated with aging in PMW (P = 0.005). CONCLUSIONS: In studies that isolated the pituitary from endogenous GnRH stimulation, aging attenuated the LH and FSH responses to exogenous GnRH in PMW. These studies indicate that the pituitary plays a role in the decline in gonadotropin levels with aging, further supporting the potential contribution of age-associated changes in both hypothalamic and pituitary function to reproductive senescence. PMID- 19549741 TI - Bone mineral density, prevalence of vertebral fractures, and bone quality in patients with adrenal incidentalomas with and without subclinical hypercortisolism: an Italian multicenter study. AB - CONTEXT: In patients with adrenal incidentalomas and subclinical hypercortisolism (SH), the factors influencing bone and the prevalence of vertebral fractures are debated. Spinal deformity index (SDI), which reflects bone quality, has never been evaluated. OBJECTIVE: The objective of the study was to investigate in these patients SDI and factors influencing the prevalence of fractures. DESIGN: This was a retrospective, multicenter study. SETTING: The study was conducted on an in and outpatient basis. PATIENTS: Patients included 287 adrenal incidentaloma patients (111 eugonadal males, 31 premenopausal, 145 postmenopausal females) and 194 controls (90 eugonadal males, 29 premenopausal, 75 postmenopausal females). MAIN OUTCOME MEASURE: Bone mineral density (BMD) was measured by dual X-ray absorptiometry at lumbar spine and femoral neck. By radiograph each vertebra was assessed as intact (grade 0) or grade 1 (20-25%), 2 (25-40%), or 3 (>40%) deformity; SDI was calculated by summing the grade of deformity for each vertebra. SH was diagnosed in the presence of at least two of the following: urinary free cortisol greater than 70 microg per 24 h (193.1 nmol/liter), cortisol after 1-mg dexamethasone test greater than 3.0 microg/dl (>82.8 nmol/liter), ACTH less than 10 pg/ml (<2.2 pmol/liter). RESULTS: BMD was significantly lower in SH+ than SH- patients and controls (lumbar spine -0.73 +/- 1.43, 0.17 +/- 1.33, 0.12 +/- 1.21, respectively; femoral neck -0.37 +/- 1.06, 0.07 +/- 1.09, 0.17 +/- 1.02). Patients with SH had higher fracture prevalence and SDI than those without SH and controls (70.6, 22.2, 21.8%, respectively, P < 0.0001; 0.31 +/- 0.68, 0.39 +/- 0.93, 1.35 +/- 1.27, respectively, P < 0.0001). Fractures and SDI were associated with SH (odds ratio 7.27, 95% confidence interval 3.94-13.41, P = 0.0001; beta = 0.352, t = 6.241, P = 0.0001, respectively) regardless of age, BMD, menopause, and gender. CONCLUSION: SH is associated with low BMD, high fracture prevalence, and reduced bone quality as measured by SDI. PMID- 19549742 TI - Threshold for effects of vitamin D deficiency on glucose metabolism in obese female African-American adolescents. AB - CONTEXT: Vitamin D status can influence insulin resistance. OBJECTIVE: The aim of the study was to determine the prevalence of vitamin D deficiency in obese African-American (AA) adolescent females in a southeastern latitude and to determine the relationship of 25-hydroxyvitamin D [25(OH)D] with insulin and glucose dynamics. DESIGN: We conducted a cross-sectional study in a University Children's Hospital. METHODS: Serum 25(OH)D, fasting glucose, PTH, serum calcium, serum lipids, serum transaminases, and C-reactive protein were assessed. Indices of insulin sensitivity and resistance were determined from an oral glucose tolerance test. Subjects were classified as vitamin D deficient or sufficient, based on the traditional vitamin D deficiency definition [serum 25(OH)D <20 ng/ml] and also by a lower 25(OH)D cut-point of 15 ng/ml or less. RESULTS: A total of 51 AA adolescent females (body mass index, 43.3 +/- 9.9 kg/m(2); age, 14 +/- 2 yr) were studied. Serum 25(OH)D concentrations were 20 ng/ml or less in 78.4% and 15 ng/ml or less in 60.8% of subjects. There were no significant group differences in the metabolic outcomes when subjects were classified using the traditional vitamin D deficiency definition. The Matsuda index of insulin sensitivity was significantly lower (P = 0.02), and insulin area under the curve was significantly higher (P = 0.04) in subjects with 25(OH)D concentrations of 15 ng/ml or less vs. those with higher concentrations. CONCLUSIONS: Vitamin D deficiency is highly prevalent in obese, AA female adolescents and may promote insulin resistance. Our data suggest that a 25(OH)D concentration of 15 ng/ml or less may be the threshold by which vitamin D deficiency confers negative effects on insulin sensitivity. PMID- 19549743 TI - Growth hormone receptor antagonist treatment reduces exercise performance in young males. AB - CONTEXT: The effects of GH on exercise performance remain unclear. OBJECTIVE: The aim of the study was to examine the effects of GH receptor (GHR) antagonist treatment on exercise performance. DESIGN: Subjects were treated with the GHR antagonist pegvisomant or placebo for 16 d. After the treatment period, they exercised to determine exercise performance and hormonal and metabolic responses. PARTICIPANTS: Twenty healthy males participated in the study. INTERVENTION: Subjects were treated with the GHR antagonist (n = 10; 10 mg/d) or placebo (n = 10). After the treatment period, they performed a maximal oxygen uptake (VO(2 max)) test and a prolonged exercise test, consisting of 60 min of submaximal cycling followed by exercise to fatigue at 90% of VO(2 max). MAIN OUTCOME MEASURES: VO(2 max) was measured before and after the treatment period. Hormonal and metabolic responses and time to exhaustion during prolonged exercise were determined. RESULTS: Resting serum IGF-I concentration decreased by 20% in the GHR antagonist-treated group (P < 0.05), whereas no change was observed in the placebo group. Conversely, resting serum GH concentration was significantly higher in the treatment group compared with the placebo group (P < 0.01). VO(2 max) did not change significantly in either group after the treatment period. Time to exhaustion at 90% of VO(2 max) was significantly shorter in the treatment group (P < 0.05). No significant differences were observed between the groups in terms of changes in serum free fatty acids, glycerol, VO(2), or relative fat oxidation. CONCLUSION: GH might be an important determinant of exercise capacity during prolonged exercise, but GHR antagonist did not alter fat metabolism during exercise. PMID- 19549744 TI - Thyroid hormone-related regulation of gene expression in human fatty liver. AB - CONTEXT: Fatty liver is an important complication of obesity; however, regulatory mechanisms mediating altered gene expression patterns have not been identified. OBJECTIVE: The aim of the study was to identify novel transcriptional changes in human liver that could contribute to hepatic lipid accumulation and associated insulin resistance, type 2 diabetes, and nonalcoholic steatohepatitis. DESIGN: We evaluated gene expression in surgical liver biopsies from 13 obese (nine with type 2 diabetes) and five control subjects using Affymetrix U133A microarrays. PCR validation was performed in liver biopsies using an additional 16 subjects. We also tested thyroid hormone responses in mice fed chow or high-fat diet. SETTING: Recruitment was performed in an academic medical center. PARTICIPANTS: Individuals undergoing elective surgery for obesity or gallstones participated in the study. RESULTS: The top-ranking gene set, down-regulated in obese subjects, was comprised of genes previously demonstrated to be positively regulated by T(3) in human skeletal muscle (n = 399; P < 0.001; false discovery rate = 0.07). This gene set included genes related to RNA metabolism (SNRPE, HNRPH3, TIA1, and SFRS2), protein catabolism (PSMA1, PSMD12, USP9X, IBE2B, USP16, and PCMT1), and energy metabolism (ATP5C1, COX7C, UQCRB). We verified thyroid hormone regulation of these genes in the liver after injection of C57BL/6J mice with T(3) (100 microg/100 g body weight); furthermore, T(3)-induced increases in expression of these genes were abolished by high-fat diet. In agreement, expression of these genes inversely correlated with liver fat content in humans. CONCLUSIONS: These data suggest that impaired thyroid hormone action may contribute to altered patterns of gene expression in fatty liver. PMID- 19549745 TI - Insulin-stimulated translocation of glucose transporter (GLUT) 12 parallels that of GLUT4 in normal muscle. AB - CONTEXT: GLUT4 is the predominant glucose transporter isoform expressed in fat and muscle. In GLUT4 null mice, insulin-stimulated glucose uptake into muscle was diminished but not eliminated, suggesting that another insulin-sensitive system was present. OBJECTIVE: This study was intended to determine whether insulin caused GLUT12 translocation in muscle. DESIGN: Six normal volunteers had muscle biopsies before and after euglycemic insulin infusions. SETTING: Infusions and biopsies were performed in an outpatient clinic. PARTICIPANTS: Subjects were nonobese, young adults with no family history of diabetes. MAIN OUTCOME MEASURES: GLUT12, GLUT4, and GLUT1 proteins were quantified in muscle biopsy fractions. Cultured myoblasts were used to determine whether GLUT12 translocation was phosphatidyl inositol-3 kinase (PI3-K)-dependent. INTERVENTION: Insulin was infused at 40 mU/m(2) x min for 3 h. RESULTS: In human muscle, insulin caused a shift of a portion of GLUT12 from intracellular low-density microsomes to the plasma membrane (PM) fraction (17% in PM at baseline, 38% in PM after insulin). Insulin increased GLUT4 in PM from 13 to 42%. GLUT1 was predominantly in the PM fractions at baseline and did not change significantly after insulin. L6 myoblasts in culture also expressed and translocated GLUT12 in response to insulin, but inhibiting PI3-K prevented the translocation of GLUT12 and GLUT4. CONCLUSIONS: Insulin causes GLUT12 to translocate from an intracellular location to the plasma membrane in normal human skeletal muscle. Translocation of GLUT12 in cultured myoblasts was dependent on activation of PI3-K. GLUT12 may have evolutionarily preceded GLUT4 and now provides redundancy to the dominant GLUT4 system in muscle. PMID- 19549747 TI - Effect of a warfarin adherence aid on anticoagulation control in an inner-city anticoagulation clinic population. AB - BACKGROUND: Poor adherence to warfarin therapy is a major contributor to subtherapeutic anticoagulation. OBJECTIVE: To determine whether use of a monthly medication organizer, filled at each clinic visit, improves anticoagulation control among warfarin-treated patients. METHODS: Patients who had a history of nonadherence to warfarin and were attending an inner-city anticoagulation clinic were enrolled in this prospective cohort study and provided with a 28-day medication organizer. Patients were instructed to bring their organizers and warfarin tablets to each anticoagulation clinic visit over the following 3 months. At each visit, the international normalized ratio (INR) was measured, warfarin adherence was assessed, and the organizer was filled with the prescribed warfarin regimen until the next scheduled visit. Data on warfarin adherence and INR values during the 3 months prior to enrollment were collected from medical records and compared with postenrollment data. RESULTS: Thirteen patients were enrolled and completed at least one postenrollment clinic visit. Adherence to warfarin therapy improved with use of the medication organizer (mean +/- SD preenrollment 55 +/- 25%, postenrollment 67 +/- 21%; p = 0.06). There was a significant decrease in the proportion of subtherapeutic INR values (60 +/- 25% to 35 +/- 29%; p = 0.04) and a significant improvement in the percent of time spent within the therapeutic INR range (32 +/- 22% to 56 +/- 28%; p = 0.03) after enrollment. CONCLUSIONS: Use of a monthly medication organizer significantly reduced the percent of subtherapeutic INR values and improved the time spent within the therapeutic anticoagulation range among previously nonadherent patients managed in an inner-city anticoagulation clinic. PMID- 19549746 TI - Heritability and environmental factors affecting vitamin D status in rural Chinese adolescent twins. AB - CONTEXT: Factors associated with the high prevalence of vitamin D deficiency in China are not well described, especially among Chinese adolescents. OBJECTIVES: The aim of the study was to examine important environmental or sociodemographic factors influencing 25-hydroxyvitamin D [25(OH)D] levels and estimate its heritability. DESIGN: A sample of 226 male and female adolescent twins aged 13-20 yr from a large prospective twin cohort of rural Chinese children and adolescents that has been followed for 6 yr were evaluated. MAIN OUTCOME MEASURE(S): Blood level of 25(OH)D was measured using tandem mass spectrometry methodology. RESULTS: The overall mean (SD) 25(OH)D level was 18.0 (9.4) ng/ml, with wide variation by gender and season. In males (47.4% of subjects), the mean (SD) 25(OH)D level was 12.1 (4.2) ng/ml in non-summer and 27.4 (8.8) ng/ml in summer; in females, it was 10.1 (4.1) ng/ml in non-summer and 19.5 (6.3) ng/ml in summer. A multivariate model that included gender, age, season, physical activity, and student status demonstrated that male gender, summer season, and high physical activity significantly increased 25(OH)D levels. Summer season and male gender also significantly decreased the risk of being in the lowest 25(OH)D tertile. Overall, 68.9% of the variability in 25(OH)D level was attributable to additive genetic influence. Stratification by gender found that in males, 85.9% of the variability in 25(OH)D level was attributable to such influence, but in females, it was only 17%. CONCLUSION: In this sample of rural Chinese adolescents, 25(OH)D level was influenced by gender, season, and physical activity level. There was a strong genetic influence on 25(OH)D level in males only. PMID- 19549748 TI - Lack of regulation of 11beta-hydroxysteroid dehydrogenase type 1 during short term manipulation of GH in patients with hypopituitarism. AB - OBJECTIVE: Evidence from long-term clinical studies measuring urinary steroid ratios, and from in vitro studies, suggests that GH administered for longer than 2 months down-regulates 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1), thereby reducing cortisol regeneration in liver and adipose tissue. We aimed to measure acute effects of GH on 11beta-HSD1 in liver and adipose tissue in vivo, including using a stable isotope tracer. DESIGN: Observational studies of GH withdrawal and reintroduction in patients with hypopituitarism. METHODS: Twelve men with benign pituitary disease causing GH and ACTH deficiency on stable replacement therapy for >6 months were studied after GH withdrawal for 3 weeks, and after either placebo or GH injections were reintroduced for another 3 weeks. We measured cortisol kinetics during 9,11,12,12-(2)H(4)-cortisol (d4-cortisol) infusion, urinary cortisol/cortisone metabolite ratios, liver 11beta-HSD1 by appearance of plasma cortisol after oral cortisone, and 11beta-HSD1 mRNA levels in subcutaneous adipose biopsies. RESULTS: GH withdrawal and reintroduction had no effect on 9,12,12-[(2)H](3)-cortisol (d3-cortisol) appearance, urinary cortisol/cortisone metabolite ratios, initial appearance of cortisol after oral cortisone, or adipose 11beta-HSD1 mRNA. GH withdrawal increased plasma cortisol 30-180 min after oral cortisone, increased d4-cortisol clearance, and decreased relative excretion of 5alpha-reduced cortisol metabolites. CONCLUSIONS: In this setting, GH did not regulate 11beta-HSD1 rapidly in vivo in humans. Altered cortisol metabolism with longer term changes in GH may reflect indirect effects on 11beta-HSD1. These data do not suggest that glucocorticoid replacement doses need to be increased immediately after introducing GH therapy to compensate for reduced 11beta-HSD1 activity, although dose adjustment may be required in the longer term. PMID- 19549749 TI - Antenatal micronutrient supplementation reduces metabolic syndrome in 6- to 8 year-old children in rural Nepal. AB - Previously, we showed that antenatal micronutrient supplementation increases birth weight in a malnourished rural South Asian setting, but the long-term effects are unknown. Between 1999 and 2001, pregnant women were sector-randomized to receive from early pregnancy through 3 mo postpartum daily micronutrient supplements containing either vitamin A alone as the control or with folic acid; folic acid+iron; folic acid+iron+zinc; or a multiple micronutrient supplement that included the above nutrients plus 11 others. From 2006 to 2008, 3524 children (93% of surviving children) were revisited between the ages of 6 and 8 y. Blood pressure, BMI, waist circumference, glycated hemoglobin, cholesterol, triglycerides, glucose, insulin, and the urinary microalbumin:creatinine ratio were assessed among children. Insulin resistance was estimated using the homeostasis model assessment (HOMA) and metabolic syndrome was defined using a modified National Cholesterol Education Program definition. None of the micronutrient supplement combinations affected blood pressure, cholesterol, triglycerides, glucose, insulin, or HOMA. There was a reduced risk of microalbuminuria (> or =3.40 mg/mmol creatinine) in the folic acid [odds ratio (OR), 0.56; 95%CI, 0.33-0.93; P = 0.02) and folic acid+iron+zinc (OR, 0.53; CI, 0.32-0.89; P = 0.02) groups and a reduced risk of metabolic syndrome in the folic acid group (OR, 0.63; CI, 0.41-0.97; P = 0.03). Maternal supplementation with folic acid or folic acid+iron+zinc reduced the risk of kidney dysfunction and, to some extent, metabolic syndrome among children at 6-8 y of age. Supplementation with multiple micronutrients had no such affect. Future follow-up studies are needed to examine long-term supplementation effects on risk of chronic diseases in adults. PMID- 19549750 TI - Lactating porcine mammary tissue catabolizes branched-chain amino acids for glutamine and aspartate synthesis. AB - The uptake of branched-chain amino acids (BCAA) from plasma by lactating porcine mammary gland substantially exceeds their output in milk, whereas glutamine output is 125% greater than its uptake from plasma. In this study, we tested the hypothesis that BCAA are catabolized for glutamine synthesis in mammary tissue. Mammary tissue slices from sows on d 28 of lactation were incubated at 37 degrees C for 1 h in Krebs buffer containing 0.5 or 2 mmol/L l-[1-(14)C]- or l-[U-(14)C] labeled leucine, isoleucine, or valine. Rates of BCAA transport and degradation in mammary tissue were high, with approximately 60% of transaminated BCAA undergoing oxidative decarboxylation and the remainder being released as branched chain alpha-ketoacids (BCKA). Most ( approximately 70%) of the decarboxylated BCAA were oxidized to CO(2). Rates of net BCAA transamination were similar to rates of glutamate, glutamine, aspartate, asparagine, and alanine synthesis. Consistent with the metabolic data, mammary tissue expressed BCAA aminotransferase (BCAT), BCKA decarboxylase, glutamine synthetase (GS), glutamate oxaloacetate aminotransferase, glutamate-pyruvate aminotransferase, and asparagine synthetase, but no phosphate-activated glutaminase, activity. Western blot analysis indicated relatively high levels of mitochondrial and cytosolic isoforms of BCAT, as well as BCKA dehydrogenase and GS proteins in mammary tissue. Our results demonstrate that glutamine and aspartate (abundant amino acids in milk protein) were the major nitrogenous products of BCAA catabolism in lactating porcine mammary tissue and provide a biochemical basis to explain an enrichment of glutamine and aspartate in sow milk. PMID- 19549751 TI - Daily injection of tumor necrosis factor-{alpha} increases hepatic triglycerides and alters transcript abundance of metabolic genes in lactating dairy cattle. AB - To determine whether inflammation can induce bovine fatty liver, we administered recombinant bovine tumor necrosis factor-alpha (rbTNF) to late-lactation Holstein cows. Cows (n = 5/treatment) were blocked by feed intake and parity and randomly assigned within block to control (CON; saline), rbTNF at 2 microg/(kg.d), or pair fed control (saline, intake matched) treatments. Treatments were administered once daily by subcutaneous injection for 7 d. Plasma samples were collected daily for analysis of glucose and FFA and a liver biopsy was collected on d 7 for triglyceride (TG) and quantitative RT-PCR analyses. Data were analyzed using treatment contrasts to assess effects of tumor necrosis factor-alpha (TNFalpha) and decreased feed intake. By d 7, feed intake of both rbTNF and pair-fed cows was approximately 15% less than CON (P < 0.01). Administration of rbTNF resulted in greater hepatic TNFalpha mRNA and protein abundance and 103% higher liver TG content (P < 0.05) without affecting the plasma FFA concentration. Hepatic carnitine palmitoyltransferase 1 transcript abundance tended to be lower (P = 0.09) and transcript abundance of fatty acid translocase and 1-acyl-glycerol-3 phosphate acyltransferase was higher (both P < 0.05) after rbTNF treatment, consistent with increased FFA uptake and storage as TG. Transcript abundance of glucose-6-phosphatase (P < 0.05) and phosphoenolpyruvate carboxykinase 1 (P = 0.09), genes important for gluconeogenesis, was lower for rbTNF-treated cows. These findings indicate that TNFalpha promotes liver TG accumulation and suggest that inflammatory pathways may also be responsible for decreased glucose production in cows with fatty liver. PMID- 19549752 TI - Successful manipulation of the quality and quantity of fat and carbohydrate consumed by free-living individuals using a food exchange model. AB - Our objective in this study was to develop and implement an effective intervention strategy to manipulate the amount and composition of dietary fat and carbohydrate (CHO) in free-living individuals in the RISCK study. The study was a randomized, controlled dietary intervention study that was conducted in 720 participants identified as higher risk for or with metabolic syndrome. All followed a 4-wk run-in reference diet [high saturated fatty acids (SF)/high glycemic index (GI)]. Volunteers were randomized to continue this diet for a further 24 wk or to 1 of 4 isoenergetic prescriptions [high monounsaturated fatty acids (MUFA)/high GI; high MUFA/low GI; low fat (LF)/high GI; and LF/low GI]. We developed a food exchange model to implement each diet. Dietary records and plasma phospholipid fatty acids were used to assess the effectiveness of the intervention strategy. Reported fat intake from the LF diets was significantly reduced to 28% of energy (%E) compared with 38%E from the HM and LF diets. SF intake was successfully decreased in the HM and LF diets to < or =10%E compared with 17%E in the reference diet (P = 0.001). Dietary MUFA in the HM diets was approximately 17%E, significantly higher than in the reference (12%E) and LF diets (10%E) (P = 0.001). Changes in plasma phospholipid fatty acids provided further evidence for the successful manipulation of fat intake. The GI of the HGI and LGI arms differed by approximately 9 points (P = 0.001). The food exchange model provided an effective dietary strategy for the design and implementation across multiple sites of 5 experimental diets with specific targets for the proportion of fat and CHO. PMID- 19549754 TI - Maternal, infant, and household factors are associated with breast-feeding trajectories during infants' first 6 months of life in Matlab, Bangladesh. AB - Women's breast-feeding patterns are complex, and existing definitions of breast feeding behavior do not capture this complexity adequately. We used results from a prior qualitative study to define trajectories for feeding during the first half of infancy, and then examined household-, maternal-, and infant-level determinants of these trajectories using logistic regression analysis. The 1472 women in the study cohort lived in rural Bangladesh and were participants in the Maternal and Infant Nutrition Intervention in Matlab trial. The 3 infant feeding trajectories included women who fed only breast milk and water [full breast feeding trajectory (FBT)]; offered mixed feeding continuously when their babies were 0-4 mo old [continuous mixed feeding trajectory (CMFT)]; and practiced any other type of breast-feeding [intermittent feeding trajectory (IFT)], which was the normative feeding behavior in this community. In adjusted regression models, women who lived in rural areas [odds ratio (OR), 2.1; 95% CI, 1.2, 3.4], came from the poorest households (OR, 3.4; 95% CI, 1.5, 7.7), and offered prelacteal (OR, 1.7; 95% CI, 1.0-2.7) were more likely to be in the FBT. Women from the richest households (OR, 1.6; 95% CI, 1.1, 2.6), employed mothers (OR, 1.7; 95% CI, 1.1, 2.6), and older mothers (OR, 1.02; 95% CI, 1.0, 1.1) were more likely to be in the CMFT, and women with higher birth-weight infants (OR, 0.6; 95% CI, 0.4 0.8) were less likely to be in the CMFT. Thus, these trajectories were associated with distinct groups of women and these results provide information useful for developing interventions to improve breast-feeding practices. PMID- 19549753 TI - Associations of gestational exposure to famine with energy balance and macronutrient density of the diet at age 58 years differ according to the reference population used. AB - Individuals exposed to the Dutch Famine of 1944-45 during gestation have increased adiposity, which might be due to changes in energy intake, physical activity, or metabolic efficiency. We studied 357 persons born between January 1945 and March 1946 whose mothers experienced famine during or immediately preceding pregnancy, 298 persons born in the same 3 institutions during 1943 or 1947 (time controls), and 311 same-sex sibling controls. We obtained food frequency and physical activity data by questionnaire between 2003 and 2005 (mean age 58 y). We defined gestational exposure as exposure to a ration of <3762 kJ/d (<900 kcal/d) for at least 10 wk. For the whole study population, energy intake was 9225 +/- 2650 kJ/d and physical activity was 7380 +/- 4331 metabolic equivalents (MET).min/wk. Compared with time controls, gestational famine exposure was associated with 113 kJ/d (95% CI, -272, 502) higher energy intake, 0.01 percentage point (95% CI, -0.88, 0.89) higher fat density, 688 MET.min/wk (95% CI, -1398, 23) lower physical activity, and 63 kJ/d (95% CI, -130, 259) higher predicted energy expenditure (pEE). Compared with sibling controls, gestational famine exposure was associated with 4 kJ/d (95% CI, -702, 711) higher energy intake, 2.01 percentage points (95% CI, 0.38, 3.63) higher fat density, 97 MET.min/wk) (95% CI, -1243, 1050) lower physical activity score, and 188 kJ/d (95% CI, -163, 539) higher pEE. Gender-specific associations (P < 0.05 for heterogeneity) emerged for protein density and pEE using time controls and for energy intake using sibling controls. Associations were weak, differed by choice of control, and may reflect sampling variability or methodological differences. Persistent small energy imbalances could explain the increased weight of famine exposed individuals. PMID- 19549755 TI - Probiotic preparation VSL#3 alters the distribution and phenotypes of dendritic cells within the intestinal mucosa in C57BL/10J mice. AB - Probiotic nutrients have shown promise in therapy for the treatment of gastrointestinal inflammation, infection, and atopic disease. Intestinal dendritic cells (DC) play a critical role in shaping the intestinal immune response. In this study, we tested the effect of a probiotic preparation (VSL#3) on DC distribution and phenotypes within the intestinal mucosa using a lineage depletion-based flow cytometric analysis. In naive C57BL/10J mice, intestinal mucosal DC were composed of plasmacytoid DC (pDC) and myeloid DC (mDC). The pDC were the dominant form in lamina propria and Peyer's patches, whereas mDC were the prevailing type in the mesenteric lymph nodes. Additional characterization of pDC and mDC with flow cytometry revealed that they expressed heterogeneous phenotypes in the intestinal mucosa. In mice gavaged with the probiotic VSL#3 for 7 d, the proportion of pDC within the lamina propria was >60% lower, whereas the pDC subset in the mesenteric lymph nodes was more than 200% greater than in sham treated controls (P < 0.01). Within pDC, the proportion of functionally unique CX3CR1(+) DC was greater than in controls in both the lamina propria and the Peyer's patches (P < 0.01). In contrast to pDC, the mDC number was greater than in controls in all intestinal lymphoid tissue compartments in VSL#3-treated mice (P < 0.01). In conclusion, this study suggests that phenotypically and functionally distinct DC subsets are localized to specific lymphoid tissues within the intestinal mucosa and that the VSL#3 probiotic nutritional supplement alters the distribution of the DC subsets within the intestinal mucosa. These changes may be important in the alteration of mucosal immunity following probiotic VSL#3 therapy. PMID- 19549757 TI - Endocannabinoids may mediate the ability of (n-3) fatty acids to reduce ectopic fat and inflammatory mediators in obese Zucker rats. AB - Dietary (n-3) long-chain PUFA [(n-3) LCPUFA] ameliorate several metabolic risk factors for cardiovascular diseases, although the mechanisms of these beneficial effects are not fully understood. In this study, we compared the effects of dietary (n-3) LCPUFA, in the form of either fish oil (FO) or krill oil (KO) balanced for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) content, with a control (C) diet containing no EPA and DHA and similar contents of oleic, linoleic, and alpha-linolenic acids, on ectopic fat and inflammation in Zucker rats, a model of obesity and related metabolic dysfunction. Diets were fed for 4 wk. Given the emerging evidence for an association between elevated endocannabinoid concentrations and metabolic syndrome, we also measured tissue endocannabinoid concentrations. In (n-3) LCPUFA-supplemented rats, liver triglycerides and the peritoneal macrophage response to an inflammatory stimulus were significantly lower than in rats fed the control diet, and heart triglycerides were lower, but only in KO-fed rats. These effects were associated with a lower concentration of the endocannabinoids, anandamide and 2 arachidonoylglycerol, in the visceral adipose tissue and of anandamide in the liver and heart, which, in turn, was associated with lower levels of arachidonic acid in membrane phospholipids, but not with higher activity of endocannabinoid degrading enzymes. Our data suggest that the beneficial effects of a diet enriched with (n-3) LCPUFA are the result of changes in membrane fatty acid composition. The reduction of substrates for inflammatory molecules and endocannabinoids may account for the dampened inflammatory response and the physiological reequilibration of body fat deposition in obese rats. PMID- 19549756 TI - Fish oil-fed mice have impaired resistance to influenza infection. AB - Dietary fish oils, rich in (n-3) PUFA, including eicosapentaenoic acid and docosahexaenoic acid, have been shown to have antiinflammatory properties. Although the antiinflammatory properties of fish oil may be beneficial during a chronic inflammatory illness, the same antiinflammatory properties can suppress the inflammatory responses necessary to combat acute viral infection. Given that (n-3) fatty acid-rich fish oil supplementation is on the rise and with the increasing threat of an influenza pandemic, we tested the effect of fish oil feeding for 2 wk on the immune response to influenza virus infection. Male C57BL/6 mice fed either a menhaden fish oil/corn oil diet (4 g fish oil:1 g corn oil, wt:wt at 5 g/100 g diet) or a control corn oil diet were infected with influenza A/PuertoRico/8/34 and analyzed for lung pathology and immune function. Although fish oil-fed mice had lower lung inflammation compared with controls, fish oil feeding also resulted in a 40% higher mortality rate, a 70% higher lung viral load at d 7 post infection, and a prolonged recovery period following infection. Although splenic natural killer (NK) cell activity was suppressed in fish oil-fed mice, lung NK activity was not affected. Additionally, lungs of infected fish oil-fed mice had significantly fewer CD8+ T cells and decreased mRNA expression of macrophage inflammatory protein-1-alpha, tumor necrosis factor alpha, and interleukin-6. These results suggest that the antiinflammatory properties of fish oil feeding can alter the immune response to influenza infection, resulting in increased morbidity and mortality. PMID- 19549758 TI - Hepcidin decreases iron transporter expression in vivo in mouse duodenum and spleen and in vitro in THP-1 macrophages and intestinal Caco-2 cells. AB - Hepcidin is thought to control iron metabolism by interacting with the iron efflux transporter ferroportin. In macrophages, there is compelling evidence that hepcidin directly regulates ferroportin protein expression. However, the effects of hepcidin on intestinal ferroportin levels are less conclusive. In this study, we compared the effects of hepcidin on iron transporter expression in the spleen and duodenum of mice treated with hepcidin over a 24- to 72-h period and observed a marked decrease in the expression of ferroportin in both duodenal enterocytes and splenic macrophages following treatment. Changes in transporter protein expression were associated with significant decreases in duodenal iron transport and serum iron. In THP-1 macrophages, ferroportin protein levels were decreased by 300 and 1000 nmol/L hepcidin. In contrast, ferroportin protein expression was unaltered in intestinal Caco-2 cells following exposure to hepcidin. However, iron efflux from Caco-2 cells was significantly inhibited in the presence of hepcidin, suggesting that the peptide could block ferroportin function in these cells. We conclude that hepcidin regulates the release of iron from both enterocytes and macrophages. However, taken together with our previous work, it is apparent that macrophages are more sensitive than enterocytes to a hepcidin challenge. PMID- 19549759 TI - Development and validation of the nutrient-rich foods index: a tool to measure nutritional quality of foods. AB - Ranking and/or classifying foods based on their nutrient composition is known as nutrient profiling. Nutrition quality indices need to be tested and validated against quality of the total diet. A family of nutrient-rich foods (NRF) indices were validated against the Healthy Eating Index (HEI), an accepted measure of diet quality. All foods consumed by participants in NHANES 1999-2002 studies were scored using NRFn.3 (where n = 6-15) indices based on unweighted sums, means, and ratios of percent daily values (DV) for nutrients to encourage (n) and for nutrients to limit (LIM) (3). Individual food scores were calculated based on 100 kcal (418 kJ) and FDA serving sizes [reference amounts customarily consumed (RACC)]. Energy-weighted food-based scores per person were then regressed against HEI, adjusting for gender, age, and ethnicity. The measure of index performance was the percentage of variation in HEI (R2) explained by each NRF score. NRF indices based on both nutrients to encourage and LIM performed better than indices based on LIM only. Maximum variance in HEI was explained using 6 or 9 nutrients to encourage; index performance actually declined with the inclusion of additional vitamins and minerals. NRF indices based on 100 kcal (418 kJ) performed similarly to indices based on RACC. Algorithms based on sums or means of nutrient DV performed better than ratio-based scores. The NRF9.3 index, based on 9 nutrients to encourage and 3 LIM per RACC and per 100 kcal, explained the highest percentage of variation from HEI and could be readily expected to rank foods based on nutrient density. PMID- 19549760 TI - Oral leucine enhances myocardial protein synthesis in rats acutely administered ethanol. AB - Acute alcohol ingestion induces an inhibition of myocardial protein synthesis by impairing mRNA translation initiation. Elevating plasma leucine (Leu) concentrations via oral gavage stimulates mRNA translation initiation in several tissues, although the effect in heart has not been well defined. The experiments described herein were designed to test the effects of a gavage solution containing Leu on protein synthesis and potential mechanisms important in accelerating mRNA translation initiation in cardiac muscle of rats given ethanol acutely to mimic "binge" dinking. Gavage with Leu stimulated protein synthesis and enhanced the assembly of the active eukaryotic initiation factor (eIF)4G.eIF4E complex. Increased assembly of the active eIF4G.eIF4E complex was associated with a 130% rise in phosphorylation of eIF4G(Ser(1108)) and a decreased assembly ( approximately 30%) of inactive eIF4E-binding protein1 (4EBP1).eIF4E complex in rats-administered ethanol. The reduced assembly of the 4EBP1.eIF4E complex was associated with an increase in phosphorylation of 4EBP1 in the hyperphosphorylated gamma-form following Leu gavage. Phosphorylation of mammalian target of rapamycin on Ser(2448), an upstream regulator of phosphorylation of 4EBP1, was elevated following Leu gavage. Neither the phosphorylation of 70-kDa ribosomal protein S6 kinase on Thr(389) nor eIF4E phosphorylation was increased following Leu gavage under any condition. Leu gavage accelerates myocardial protein synthesis following acute ethanol intoxication by enhancing eIF4G.eIF4E complex assembly through increased phosphorylation of eIF4G and decreased association of 4EBP1 with eIF4E. PMID- 19549761 TI - Resveratrol alters proliferative responses and apoptosis in human activated B lymphocytes in vitro. AB - We hypothesized that the phytochemicals resveratrol, quercetin, and kaempferol would modulate B lymphocyte proliferation, Ig synthesis, and apoptosis after activation. Peripheral blood mononuclear cells (PBMC) were isolated from 12 healthy adult human volunteers and incubated with pokeweed mitogen plus 0, 2, 5, and 10 mumol/L resveratrol, quercetin, or kaempferol. After 6 d, CD19+ B cells were analyzed for proliferation, B cell lymphoma-2 (Bcl-2) expression, and activation of caspase-3 using flow cytometry. After 8 d, cell supernatants were collected and IgM and IgG were measured by ELISA. Resveratrol at a concentration of 5 mumol/L increased the percentage of CD19+ cells compared with mitogen only stimulated cells (P < 0.01), and a trend for increased proliferation was observed for cells treated with 0, 2, and 5 mumol/L resveratrol (P-trend = 0.01). However, 10 mumol/L resveratrol inhibited proliferation of B lymphocytes (P < 0.01). Expression of Bcl-2 and caspase-3 activation increased in B cells treated with 10 mumol/L resveratrol compared with mitogen alone (P < 0.01), and trends for dose responsive increases in Bcl-2 expression and caspase-3 activation were observed (P-trend < 0.0001). Differences in IgM and IgG production were not observed for PBMC treated with resveratrol. Kaempferol at 10 mumol/L slightly inhibited proliferative responses (P < 0.05) but did not affect B cell function or apoptosis. Quercetin did not alter B cell proliferation, function, or apoptosis. These data show that human B lymphocyte proliferation and apoptosis are modified by physiological concentrations of resveratrol and suggest that exposure of human B cells to resveratrol may increase survival by upregulating Bcl-2. PMID- 19549762 TI - Up-regulation of AGS3 during morphine withdrawal promotes cAMP superactivation via adenylyl cyclase 5 and 7 in rat nucleus accumbens/striatal neurons. AB - Effective medical treatment of opiate addiction is limited by a high relapse rate in abstinent addicts. Opiate withdrawal causes cAMP superactivation, but the underlying molecular mechanisms are not clear. Recent evidence implicates an activator of G-protein signaling 3 (AGS3) in opiate addiction. We found previously that during a 10-min activation of opioid receptors, AGS3 binds G alpha(i)-GDP to promote free G betagamma stimulation of adenylyl cyclase (AC) 2 and 4, and/or inactivate G alpha(i) inhibitory function, thereby transiently enhancing cAMP-dependent protein kinase A (PKA) activity. In contrast, we report here that in nucleus accumbens/striatal neurons, morphine withdrawal induces cAMP superactivation, which requires up-regulation of AGS3. cAMP increases as a function of withdrawal time, by approximately 20% at 10 min and 75% at 5 h. However, cAMP superactivation does not require G betagamma. Instead, adenosine A2A receptor activation of G alpha(s/olf) seems to initiate cAMP superactivation and promote AGS3 up-regulation. Elevated AGS3 binds to G alpha(i) to prevent its inhibition on AC activation. Moreover, withdrawal-induced increases in cAMP/PKA activate phospholipase C and epsilon protein kinase C to further stimulate AC5 and AC7, causing cAMP superactivation. Our findings identify a critical role for AC 5 and 7 and A2A receptors for up-regulation of AGS3 in morphine withdrawal induced cAMP superactivation. PMID- 19549763 TI - Etoposide induces protein kinase Cdelta- and caspase-3-dependent apoptosis in neuroblastoma cancer cells. AB - In this report, we reveal that etoposide inhibits the proliferation of SK-N-AS neuroblastoma cancer cells and promotes protein kinase Cdelta (PKCdelta)- and caspase-dependent apoptosis. Etoposide induces the caspase-3-dependent cleavage of PKCdelta to its active p40 fragment, and active PKCdelta triggers the processing of caspase-3 by a positive-feedback mechanism. Treatment of cells with the caspase-3-specific inhibitor N-benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethyl ketone or caspase-3-specific small interacting RNA (siRNA) prevented the etoposide-induced activation of caspase-8 and inhibited apoptosis. The silencing of the caspase-2 or caspase-8 genes using siRNAs did not affect the etoposide induced processing of caspase-3, indicating that these caspases lie downstream of caspase-3 in this signaling pathway. Furthermore, the etoposide-induced processing of caspase-2 required the expression of caspase-8, and the etoposide mediated processing of caspase-8 required the expression of caspase-2, indicating that these two caspases activate each other after etoposide treatment. We also observed that etoposide-mediated apoptosis was decreased by treating the cells with the caspase-6-specific inhibitor benzyloxycarbonyl-Val-Glu(OMe)-Ile-Asp (OMe)-fluoromethyl ketone and that caspase-6 was activated by a caspase-8 dependent mechanism. Finally, we show that rottlerin blocks etoposide-induced apoptosis by inhibiting the PKCdelta-mediated activation of caspase-3 and by degrading caspase-2, which prevents caspase-8 activation. Our results add important insights into how etoposide mediates apoptotic signaling and how targeting these pathways may lead to the development of novel therapeutics for the treatment of neuroblastomas. PMID- 19549765 TI - What's Your Line? PMID- 19549764 TI - The role of centrosomes in mammalian fertilization and its significance for ICSI. AB - Centrosome integrity is critically important for successful fertilization and embryo development. In humans, the sperm contributes the dominant centrosomal material containing centrioles and centrosomal components onto which oocyte centrosomal proteins assemble after sperm incorporation to form the sperm aster that is essential for uniting sperm and oocyte pronuclei. Increasingly, dysfunctional sperm centrosomes have been identified as a factor for sperm derived infertility and heterologous Intracytoplasmic Sperm Injection (ICSI) has been used to assess centrosome and sperm aster formation and clearly established a relationship between infertility and sperm centrosomal dysfunction. ICSI has been used successfully to provide novel treatment to overcome male factor infertility and it may open up new possibilities to correct specific sperm related centrosome dysfunctions at molecular levels. New data indicate that it is now possible to replace dysfunctional centrosomes with functional donor sperm centrosomes which may provide new treatment for couples in which infertility is a result of centrosome-related sperm dysfunctions. PMID- 19549766 TI - Urine analysis and protein networking identify met as a marker of metastatic prostate cancer. AB - PURPOSE: Metastatic prostate cancer is a major cause of death of men in the United States. Expression of met, a receptor tyrosine kinase, has been associated with progression of prostate cancer. EXPERIMENTAL DESIGN: To investigate met as a biomarker of disease progression, urinary met was evaluated via ELISA in men with localized (n = 75) and metastatic (n = 81) prostate cancer. Boxplot analysis was used to compare the distribution of met values between each group. We estimated a receiver operating characteristic curve and the associated area under the curve to summarize the diagnostic accuracy of met for distinguishing between localized and metastatic disease. Protein-protein interaction networking via yeast two hybrid technology supplemented by Ingenuity Pathway Analysis and Human Interactome was used to elucidate proteins and pathways related to met that may contribute to progression of disease. RESULTS: Met distribution was significantly different between the metastatic group and the group with localized prostate cancer and people with no evidence of cancer (P < 0.0001). The area under the curve for localized and metastatic disease was 0.90, with a 95% confidence interval of 0.84 to 0.95. Yeast two-hybrid technology, Ingenuity Pathway Analysis, and Human Interactome identified 89 proteins that interact with met, of which 40 have previously been associated with metastatic prostate cancer. CONCLUSION: Urinary met may provide a noninvasive biomarker indicative of metastatic prostate cancer and may be a central regulator of multiple pathways involved in prostate cancer progression. PMID- 19549767 TI - Collapse of the CD27+ B-cell compartment associated with systemic plasmacytosis in patients with advanced melanoma and other cancers. AB - PURPOSE: Disturbed peripheral blood B-cell homeostasis complicates certain infections and autoimmune diseases, such as HIV and systemic lupus erythematosus, but has not been reported in cancer. This study aimed to investigate whether B cell physiology was altered in the presence of melanoma and other cancers. EXPERIMENTAL DESIGN: Flow cytometry was used to identify phenotypic differences in B cells from patients with melanoma and normal donors. In vitro stimulated B cells were assessed for responsiveness and also used as stimulators of allogeneic T cells in mixed lymphocyte reactions. RESULTS: We show B-cell dysregulation in patients with advanced melanoma (n = 26) and other solid tumors (n = 13), marked by a relative and absolute loss of CD27+ (memory) B cells and associated with an aberrant systemic plasmacytosis. Functionally, B cells from patients with melanoma inefficiently up-regulated immunoregulatory molecules and weakly secreted cytokines in response to CD40 and toll-like receptor 9 agonists. Stimulated B cells from patients induced proliferation of alloreactive CD4+ T cells, but these T cells poorly secreted IFNgamma and interleukin-2. These effects were recapitulated by using purified normal donor CD27(neg) B cells in these same assays, linking the predominance of CD27(neg) B cells in patients with the observed functional hyporesponsiveness. Indeed, B-cell dysfunction in patients strongly correlated with the extent of loss of CD27+ B cells in peripheral blood. CONCLUSIONS: Disturbed B-cell homeostasis is a previously unrecognized feature of patients with advanced melanoma and other cancers and may represent an unanticipated mechanism of immune incompetence in cancer. PMID- 19549768 TI - HPV16 tumor associated macrophages suppress antitumor T cell responses. AB - PURPOSE: High-risk human papillomavirus (HPV) is the main etiologic factor for cervical cancer. The severity of HPV-associated cervical lesions has been correlated to the number of infiltrating macrophages. The objective of this work is to characterize the role of tumor-associated macrophages (TAM) on the immune cellular response against the tumor. EXPERIMENTAL DESIGN: We used the HPV16 E6- and E7-expressing TC-1 mouse tumor model to study the effect of TAM on T-cell function in vitro, and depleted TAM, using clodronate-containing liposomes, to characterize its role in vivo. RESULTS: TAM, characterized by the positive expression of CD45, F4/80, and CD11b, formed the major population of infiltrating tumor cells. TAM displayed high basal Arginase I activity, producing interleukin 10 (IL-10); they were resistant to iNOSII activity induction, therefore reversion to M1 phenotype, when stimulated in vitro with lipopolysaccharide/IFNgamma, indicating an M2 phentoype. In cultures of isolated TAM, TAM induced regulatory phenotype, characterized by IL-10 and Foxp3 expression, and inhibited proliferation of CD8 lymphocytes. In vivo, depletion of TAM inhibited tumor growth and stimulated the infiltration of tumors by HPV16 E7(49-57)-specific CD8 lymphocytes, whereas depletion of Gr1(+) tumor-associated cells had no effect. CONCLUSIONS: M2-like macrophages infiltrate HPV16-associated tumors causing suppression of antitumor T-cell response, thus facilitating tumor growth. Depletion or phenotype alteration of this population should be considered in immunotherapy strategies. PMID- 19549770 TI - Activation of p38 mitogen-activated protein kinase drives dendritic cells to become tolerogenic in ret transgenic mice spontaneously developing melanoma. AB - PURPOSE: The purpose of the study was to investigate signaling molecules involved in the acquisition of tolerogenic properties by dendritic cells (DC) in ret transgenic mice with spontaneous melanoma progression and to target these molecules to overcome the barrier for effective melanoma immunotherapy. EXPERIMENTAL DESIGN: DC functions and expression patterns of p38 mitogen activated protein kinase (MAPK) in DCs were evaluated in a ret transgenic murine cutaneous melanoma model, which shows high similarity to human cutaneous melanoma with respect to clinical development. In contrast to transplantation melanoma models (like B16), this model allows the study of melanoma progression under conditions of natural interactions between tumor and host cells over time. RESULTS: We showed a strong tumor infiltration with immature DCs and a reduction in the number of mature DCs in lymphoid organs during melanoma progression. DCs from melanoma-bearing mice secreted significantly more interleukin 10 and less interleukin 12p70, and showed a decreased capacity to activate T cells compared with DCs from tumor-free animals. Observed DC dysfunction was linked to considerable activation of p38 MAPK. Inhibition of its activity in spleen DCs from tumor-bearing mice led to normalization of their cytokine secretion pattern and T-cell stimulation capacity. CONCLUSIONS: Our data show a critical role of constitutively activated p38 MAPK in the acquirement of tolerogenic pattern by DCs during melanoma progression that contributes to the suppression of antitumor T-cell immune responses. We suggest that new strategies of melanoma immunotherapy can include inhibitors of p38 MAPK activity in DCs. PMID- 19549769 TI - Enhanced systemic immune reactivity to a Basal cell carcinoma associated antigen following photodynamic therapy. AB - PURPOSE: Numerous preclinical studies have shown that local photodynamic therapy (PDT) of tumors enhances systemic antitumor immunity. However, other than single case and anecdotal reports, this phenomenon has not been examined following clinical PDT. To determine whether PDT in a clinical setting enhances systemic recognition of tumor cells, we examined whether PDT of basal cell carcinoma resulted in an increased systemic immune response to Hip1, a tumor antigen associated with basal cell carcinoma. EXPERIMENTAL DESIGN: Basal cell carcinoma lesions were either treated with PDT or surgically removed. Blood was collected from patients immediately before or 7 to 10 days following treatment. Peripheral blood leukocytes were isolated from HLA-A2-expressing patients and reactivity to a HLA-A2-restricted Hip1 peptide was measured by INF-gamma ELISpot assay. RESULTS: Immune recognition of Hip1 increased in patients whose basal cell carcinoma lesions were treated with PDT. This increase in reactivity was significantly greater than reactivity observed in patients whose lesions were surgically removed. Patients with superficial lesions exhibited greater enhancement of reactivity compared with patients with nodular lesions. Immune reactivity following PDT was inversely correlated with treatment area and light dose. CONCLUSIONS: These findings show for the first time that local tumor PDT can enhance systemic immune responses to tumors in patients, and validate previous preclinical findings. PMID- 19549771 TI - A phase I trial of radioimmunotherapy with 131I-A5B7 anti-CEA antibody in combination with combretastatin-A4-phosphate in advanced gastrointestinal carcinomas. AB - PURPOSE: In preclinical models, radioimmunotherapy with (131)I-A5B7 anti carcinoembryonic antigen (CEA) antibody ((131)I-A5B7) combined with the vascular disruptive agent combretastatin-A4-phosphate (CA4P) produced cures unlike either agent alone. We conducted a phase I trial determining the dose-limiting toxicity (DLT), maximum tolerated dose, efficacy, and mechanism of this combination in patients with gastrointestinal adenocarcinomas. EXPERIMENTAL DESIGN: Patients had CEA of 10 to 1,000 microg/L, QTc < or =450 ms, no cardiac arrhythmia/ischaemia, and adequate hematology/biochemistry. Tumor was suitable for blood flow analysis by dynamic contrast enhanced-magnetic resonance imaging (MRI). The starting dose was 1,800 MBq/m(2) of (131)I-A5B7 on day 1 and 45 mg/m(2) CA4P given 48 and 72 hours post-(131)I-A5B7, then weekly for up to seven weeks. RESULTS: Twelve patients were treated, with mean age of 63 years (range, 32-77). Two of six patients at the first dose level had DLTs (grade 4 neutropenia). The dose was reduced to 1,600 MBq/m(2), and CA4P escalated to 54 mg/m(2). Again, two of six patients had DLTs (neutropenia). Of ten assessable patients, three had stable disease and seven had progressive disease. Single-photon emission computed tomography confirmed tumor antibody uptake in all 10 patients. DCE-MRI confirmed falls in kinetic parameters (K(trans)/IAUGC(60)) in 9 of 12 patients. The change of both pharmacokinetic parameters reached a level expected to produce efficacy in one patient who had a minor response on computed tomography and a reduced serum tumor marker level. CONCLUSIONS: This is believed to be the first trial reporting the combination of radioimmunotherapy and vascular disruptive agent; each component was shown to function, and myelosuppression was dose-limiting. Optimal dose and timing of CA4P, and moderate improvements in the performance of radioimmunotherapy seem necessary for efficacy. PMID- 19549772 TI - Genomic screening for genes silenced by DNA methylation revealed an association between RASD1 inactivation and dexamethasone resistance in multiple myeloma. AB - PURPOSE: Epigenetic changes such as DNA methylation play a key role in the development and progression of multiple myeloma. Our aim in the present study was to use genomic screening to identify genes targeted for epigenetic inactivation in multiple myeloma and assess their role in the development of resistance to dexamethasone. EXPERIMENTAL DESIGN: Gene expression was examined using microarray screening, reverse transcription-PCR, and real-time quantitative PCR. DNA methylation was examined using bisulfite PCR, bisulfite sequencing, and bisulfite pyrosequencing in 14 multiple myeloma cell lines, 87 multiple myeloma specimens, and 12 control bone marrow samples. WST-8 assays were used to assess cell viability after treatment with 5-aza-2'-deoxycytidine and/or dexamethasone. RESULTS: Microarray analysis was done to screen for genes up-regulated by 5-aza 2'-deoxycytidine. In RPMI8226 cells, 128 genes were up-regulated, whereas 83 genes were up-regulated in KMS12PE cells. Methylation of 22 genes with CpG islands in their 5' regions, including RASD1, was confirmed. Methylation of RASD1 was associated with its inactivation, which correlated with resistance to dexamethasone. Treating multiple myeloma cells with 5-aza-2'-deoxycytidine restored sensitivity to dexamethasone. Methylation of RASD1 was also detected in a subset of primary multiple myeloma specimens, and the levels of methylation were increased after repeated antitumor treatments. Gene signature analysis revealed various genes to be synergistically induced by treatment with a combination of 5-aza-2'-deoxycytidine plus dexamethasone. CONCLUSION: Our findings indicate that epigenetic inactivation of genes, including RASD1, plays a key role in the development of dexamethasone resistance in multiple myeloma. Moreover, they show the utility of demethylation therapy in cases of advanced multiple myeloma. PMID- 19549773 TI - A cohort study of cyclin D1 expression and prognosis in 602 colon cancer cases. AB - PURPOSE: Cyclin D1 and cyclin-dependent kinases (CDK) are commonly activated in colorectal cancer. The activity of cyclin D1 can be blocked by CDK inhibitors, including p27 (CDKN1B) and p21 (CDKN1A, which is induced by p53). However, prognostic significance of tumoral cyclin D1 remains uncertain, and no previous study has considered potential confounding effect of p53, p21, p27, and related molecular events [microsatellite instability (MSI), CpG island methylator phenotype, and LINE-1 hypomethylation]. EXPERIMENTAL DESIGN: Among 602 colon cancer patients (stage I-IV) in two prospective cohort studies, cyclin D1 overexpression was detected in 330 (55%) tumors by immunohistochemistry. Cox proportional hazard models computed hazard ratios (HR) of colon cancer-specific and overall mortalities, adjusted for patient characteristics and tumoral molecular features, including p53, p21, p27, cyclooxygenase-2, fatty acid synthase, LINE-1 methylation, CpG island methylator phenotype, MSI, BMI, KRAS, and BRAF. RESULTS: Cyclin D1 overexpression was associated with a low cancer specific mortality in Kaplan-Meier analysis (P = 0.006), and in both univariate Cox regression [unadjusted HR, 0.64; 95% confidence interval (CI), 0.47-0.88; P = 0.0063] and multivariate analyses (adjusted HR, 0.57; 95% CI, 0.39-0.84; P = 0.0048). Similar findings were observed for an overall mortality (adjusted HR, 0.74; 95% CI, 0.57-0.98; P = 0.036). Notably, the effect of cyclin D1 on survival might differ by MSI status (P(interaction) = 0.008). Compared with tumors that were both cyclin D1-negative and MSI-low/microsatellite stable, the presence of either cyclin D1 or MSI-high or both seemed to confer better clinical outcome (adjusted HR point estimates, 0.10-0.65). CONCLUSIONS: Cyclin D1 overexpression is associated with longer survival in colon cancer. PMID- 19549774 TI - Detection of KRAS oncogene in peripheral blood as a predictor of the response to cetuximab plus chemotherapy in patients with metastatic colorectal cancer. AB - PURPOSE: Previously we developed membrane-arrays as a promising tool to detect circulating tumor cells (CTC) with KRAS oncogene in patients with malignancies. This study was conducted to determinate the predictive values of CTCs with KARS mutation by membrane-arrays for metastatic colorectal cancer patients treated with cetuximab plus chemotherapy. EXPERIMENTAL DESIGN: Seventy-six metastatic colorectal cancer patients receiving cetuximab plus FOLFIRI or FOLFOX-4 chemotherapy were enrolled. KRAS mutation status in the peripheral blood of these patients was analyzed using membrane-arrays, and KRAS mutation status in tumors was analyzed by DNA sequencing. RESULTS: Among 76 metastatic colorectal cancer patients, KRAS mutations in tumors and in peripheral blood were identified in 33 (43.4%) and 30 (39.5%) patients, respectively. The detection sensitivity, specificity, and accuracy of membrane-arrays for CTCs with KRAS oncogene were 84.4%, 95.3%, and 90.8%, respectively, and indeed a highly significant correlation to KRAS mutations in tumors (P < 0.0001) was observed. Forty-five (59.2%) patients responded to cetuximab plus chemotherapy, and 41 and 40 were wild-type KRAS in tumors and peripheral blood, respectively (both P < 0.0001). Patients with tumors that harbor wild-type KRAS are more likely to have a better progression-free survival and overall survival when treated with cetuximab plus chemotherapy (P < 0.0001). Likewise, patients with CTCs of wild-type KRAS in peripheral blood express a better progression-free survival and overall survival when treated with cetuximab plus chemotherapy (P < 0.0001). CONCLUSIONS: These findings provide evidence that detection of KRAS mutational status in CTCs, by gene expression array, has potential for clinical application in selecting metastatic colorectal cancer patients most likely to benefit from cetuximab therapy. PMID- 19549775 TI - Disclosure of research results from cancer genomic studies: state of the science. AB - Although the cancer research community has supported a "nondisclosure" position about the return of individual results to research subjects, new technologies, such as genome wide association studies, will reveal clinically relevant findings, some of which cannot be ignored. What recommendations exist that can guide researchers and Institutional Review Boards (IRBs) about this issue? This article summarizes the relevant public documents about the disclosure of individual research results to inform policy development. Four stakeholder groups were selected to make this comparison: federal, professional, advisory, and advocacy groups. Regardless of a group's position on disclosure, there was consensus that if research results were to be disclosed under any condition, the results must be analytically and clinically validated and that the researcher should not make this decision alone, but in conjunction with the IRB. There was no consensus, however, on the specific determinants for disclosure or what constitutes clinical validity. Although sufficient agreement exists to begin developing general guidelines about the process for disclosure of individual research results, the actual determinants with which to guide this decision remain challenging. An alternate framework that addresses the threshold of uncertainty a stakeholder is willing to accept, the positive predictive value of the research finding, and the magnitude of harm of returning results may be more effective to guide decision making. These assessments, along with what is considered useful information, requires the involvement of the research subject community to inform decision-making and move the policy process forward. PMID- 19549776 TI - Preclinical results of camptothecin-polymer conjugate (IT-101) in multiple human lymphoma xenograft models. AB - PURPOSE: Camptothecin (CPT) has potent broad-spectrum antitumor activity by inhibiting type I DNA topoisomerase (DNA topo I). It has not been used clinically because it is water-insoluble and highly toxic. As a result, irinotecan (CPT-11), a water-soluble analogue of CPT, has been developed and used as salvage chemotherapy in patients with relapsed/refractory lymphoma, but with only modest activity. Recently, we have developed a cyclodextrin-based polymer conjugate of 20-(S)-CPT (IT-101). In this study, we evaluated the preclinical antilymphoma efficacy of IT-101 as compared with CPT-11. EXPERIMENTAL DESIGN: We determined an in vitro cytotoxicity of IT-101, CPT-11, and their metabolites against multiple human lymphoma cell lines. In human lymphoma xenografts, the pharmacokinetics, inhibitions of tumor DNA topo I catalytic activity, and antilymphoma activities of these compounds were evaluated. RESULTS: IT-101 and CPT had very high in vitro cytotoxicity against all lymphoma cell lines tested. As compared with CPT-11 and SN-38, IT-101 and CPT had longer release kinetics and significantly inhibit higher tumor DNA topo I catalytic activities. Furthermore, IT-101 showed significantly prolonged the survival of animals bearing s.c. and disseminated human xenografts when compared with CPT-11 at its maximum tolerated dose in mice. CONCLUSIONS: The promising present results provide the basis for a phase I clinical trial in patients with relapsed/refractory lymphoma. PMID- 19549777 TI - Multiple Histology Phase II Trials. AB - PURPOSE: Testing agents in cancers with multiple disease subtypes, in which the activity of a new treatment may vary between subtypes, presents statistical and logistical challenges. We propose a flexible phase II strategy which includes both analyses for each histology or stratum and a combined analysis which borrows information from all the patients in the study. Sequential futility analyses are conducted once each subgroup or the overall group reaches a specified minimum accrual. EXPERIMENTAL DESIGN: Examples based on a soft tissue sarcoma phase II trial, which includes multiple histologies and simulation studies, are used to assess the statistical properties of the proposed strategy. RESULTS: The combined analyses in one phase II trial lead to smaller expected sample sizes when the drug is broadly inactive, and to greater statistical power if there is modest activity across multiple strata as compared with conducting several smaller phase II studies. In addition, by retaining the stratum-specific tests, the design allows the identification of subgroups for which the agents are most active. CONCLUSION: To consider phase II testing with multiple biological subtypes, a strategy which combines both the individual subgroup tests and overall combined tests has promising statistical properties. Our results support the appropriate use of statistical borrowing of information in phase II studies in this setting. More broadly, this work fits the paradigm that phase II studies should include as broad a group of patients as scientifically reasonable, but incorporate design considerations for subsets of patients with potentially differing responses to therapy. PMID- 19549779 TI - Engineering synthetic adaptors and substrates for controlled ClpXP degradation. AB - Facile control of targeted intracellular protein degradation has many potential uses in basic science and biotechnology. One promising approach to this goal is to redesign adaptor proteins, which can regulate proteolytic specificity by tethering substrates to energy-dependent AAA+ proteases. Using the ClpXP protease, we have probed the minimal biochemical functions required for adaptor function by designing and characterizing variant substrates, adaptors, and ClpX enzymes. We find that substrate tethering mediated by heterologous interaction domains and a small bridging molecule mimics substrate delivery by the wild-type system. These results show that simple tethering is sufficient for synthetic adaptor function. In our engineered system, tethering and proteolysis depend on the presence of the macrolide rapamycin, providing a foundation for engineering highly specific degradation of target proteins in cells. Importantly, this degradation is regulated by a small molecule without the need for new adaptor or enzyme biosynthesis. PMID- 19549778 TI - Structural and functional alterations of FLT3 in acute myeloid leukemia. AB - Hematopoiesis is highly regulated through cytokine-induced stimulation of multiple signal transduction pathways in order to mediate appropriate differentiation and proliferation of specific progenitor populations. Ligand induced stimulation of the FMS-like tyrosine kinase 3 (FLT3) leads to activation of multiple downstream effector pathways resulting in differentiation and proliferation of specific progenitor cell populations. Genomic alterations of the FLT3 gene, including FLT3 internal tandem duplication (FLT3/ITD) and FLT3 activation loop mutation (FLT3/ALM) lead to autonomous receptor activation, dysregulation of FLT3 signal transduction pathways, contribute to myeloid pathogenesis, and have been linked to response to therapy and clinical outcome. Exploring the mechanisms by which these FLT3 alterations lead to dysregulated proliferation should provide a better understanding of the molecular pathogenesis of acute myeloid leukemia (AML) and may provide insights into potential therapeutic interventions. FLT3 inhibitors are under evaluation for their efficacy in AML patients with FLT3 mutations. PMID- 19549780 TI - Coupling of ionic events to protein kinase signaling cascades upon activation of alpha7 nicotinic receptor: cooperative regulation of alpha2-integrin expression and Rho kinase activity. AB - Defining the signaling mechanisms and effector proteins mediating phenotypic and mechanical plasticity of keratinocytes (KCs) during wound epithelialization is one of the major goals in epithelial cell biology. The acetylcholine (ACh)-gated ion channels, or nicotinic ACh receptors (nAChRs), mediate the nicotinergic signaling that controls crawling locomotion of KCs. To elucidate relative contributions of the ionic and protein kinase-mediated events elicited due to activation of alpha7 nAChRs, we quantitated expression of alpha2-integrin gene at the mRNA and protein levels and also measured Rho kinase activity in KCs stimulated with the alpha7 agonist AR-R17779 while blocking the Na+ or Ca2+ entry and/or inhibiting signaling kinases. The results demonstrated the existence of the two-component signaling systems coupling the ionic events and protein kinase signaling cascades downstream of alpha7 nAChR to simultaneous up-regulation of alpha2-integrin expression and activation of Rho kinase. The Raf/MEK1/ERK1/2 cascade up-regulating alpha2-integrin was activated due to both Ca2+-dependent recruitment of Ca2+/calmodulin-dependent protein kinase II and protein kinase C and Ca2+-independent activation of Ras. Likewise the phosphatidylinositol 3 kinase-mediated activation of Rho kinase was elicited due to both Ca2+ entry dependent involvement of Ca2+/calmodulin-dependent protein kinase II and Ca2+ independent activation of Jak2. Thus, although the initial signals emanating from activated alpha7 nAChR are different in nature the pathways intersect at common effector molecules providing for a common end point effect. This novel paradigm of nAChR-mediated coordination of the ionic and metabolic signaling events can allow an auto/paracrine ACh to simultaneously alter gene expression and induce reciprocal changes in the cytoskeleton and contractile system of KCs required to compete a particular step of wound epithelialization. PMID- 19549781 TI - S-glutathionylation of the Rpn2 regulatory subunit inhibits 26 S proteasomal function. AB - Although increased intracellular concentrations of hydrogen peroxide (H2O2) are associated with inhibition of 26 S proteasomal activity, the mechanisms responsible for such effects have not been well delineated. In the present studies, we found that direct exposure of purified 26 S proteasomes to H2O2 had negligible effects on their activity, whereas incubation with glutathione and H2O2 produced >80% decrease in chymotrypsin-like and trypsin-like activities. Rpn1 and Rpn2, which are subunits of the 19 S regulatory particle, undergo S glutathionylation after exposure of purified 26 S proteasomes to glutathione and H2O2, as well as in HEK 293 cells and neutrophils incubated with H2O2. Increased oxidation of Rpn1 and Rpn2 cysteine thiols was also found in lung extracts from mice in which catalase was inactivated, a condition associated with augmented intracellular concentrations of H2O2 and diminished 26 S proteasomal activity. Although unoxidized Rpn2 enhanced 20 S proteolytic function in vitro, such potentiation was not found when the 20 S core particle was incubated with oxidized Rpn2. The composition of 26 S proteasomes was not altered after exposure to glutathione and H2O2, with similar amounts of Rpn1 and Rpn2 in control or oxidized 26 S proteasomal complexes. These findings identify S-glutathionylation of Rpn2 as a contributory mechanism for H2O2-induced inhibition of 26 S proteasomal function. PMID- 19549782 TI - Endoplasmic reticulum-associated degradation (ERAD) and autophagy cooperate to degrade polymerogenic mutant serpins. AB - The serpinopathies are a family of diseases characterized by the accumulation of ordered polymers of mutant protein within the endoplasmic reticulum. They are a diverse group including alpha(1)-antitrypsin deficiency and the inherited dementia familial encephalopathy with neuroserpin inclusion bodies or FENIB. We have used transient transfection of COS7 cells and mouse embryonic fibroblasts, PC12 cell lines that conditionally express wild type and mutant neuroserpin and fly models of FENIB to assess the cellular handling of wild type and mutant serpins. By using a polymer-specific monoclonal antibody, we show that mutant neuroserpin forms polymers after a delay of at least 30 min and that polymers can be cleared in PC12 cell lines and from the brain in a fly model of FENIB. At steady state, the fractions of intracellular polymerogenic G392E mutant neuroserpin in the monomeric and polymeric states are comparable. Inhibition of the proteasome with MG132 reveals that both mutant neuroserpin and alpha(1) antitrypsin are degraded predominantly by endoplasmic reticulum-associated degradation (ERAD). Pharmacological and genetic inhibitions demonstrate that autophagy is responsible for bulk turnover of wild type and mutant serpins, but can be stimulated by rapamycin to compensate for proteasome inhibition. The significance of these findings to the treatment of serpinopathies is discussed. PMID- 19549783 TI - Mechanism for HIV-1 Tat insertion into the endosome membrane. AB - The human immunodeficiency virus, type 1, transactivating protein Tat is a small protein that is strictly required for viral transcription and multiplication within infected cells. The infected cells actively secrete Tat using an unconventional secretion pathway. Extracellular Tat can affect different cell types and induce severe cell dysfunctions ranging from cell activation to cell death. To elicit most cell responses, Tat needs to reach the cell cytosol. To this end, Tat is endocytosed, and low endosomal pH will then trigger Tat translocation to the cytosol. Although this translocation step is critical for Tat cytosolic delivery, how Tat could interact with the endosome membrane is unknown, and the key residues involved in this interaction require identification. We found that, upon acidification below pH 6.0 (i.e. within the endosomal pH range), Tat inserts into model membranes such as monolayers or lipid vesicles. This insertion process relies on Tat single Trp, Trp-11, which is not needed for transactivation and could be replaced by another aromatic residue for membrane insertion. Nevertheless, Trp-11 is strictly required for translocation. Tat conformational changes induced by low pH involve a sensor made of its first acidic residue (Glu/Asp-2) and the end of its basic domain (residues 55-57). Mutation of one of these elements results in membrane insertion above pH 6.5. Tat basic domain is also required for efficient Tat endocytosis and membrane insertion. Together with the strict conservation of Tat Trp among different virus isolates, our results point to an important role for Tat-membrane interaction in the multiplication of human immunodeficiency virus type 1. PMID- 19549786 TI - Leflunomide and the lung. PMID- 19549785 TI - Identification of a disulfide bridge essential for transport function of the human proton-coupled amino acid transporter hPAT1. AB - The proton-coupled amino acid transporter 1 (PAT1, SLC36A1) mediates the uptake of small neutral amino acids at the apical membrane of intestinal epithelial cells after protein digestion. The transporter is currently under intense investigation, because it is a possible vehicle for oral drug delivery. Structural features of the protein such as the number of transmembrane domains, the substrate binding site, or essential amino acids are still unknown. In the present study we use mutagenesis experiments and biochemical approaches to determine the role of the three putative extracellular cysteine residues on transport function and their possible involvement in the formation of a disulfide bridge. As treatment with the reducing reagent dithiothreitol impaired transport function of hPAT1 wild type protein, substitution of putative extracellular cysteine residues Cys-180, Cys-329, and Cys-473 by alanine or serine was performed. Replacement of the two highly conserved cysteine residues Cys-180 and Cys-329 abolished the transport function of hPAT1 in Xenopus laevis oocytes. Studies of wild type and mutant transporters expressed in human retinal pigment epithelial (HRPE) cells suggested that the binding of the substrate was inhibited in these mutants. Substitution of the third putative extracellular nonconserved cysteine residue Cys-473 did not affect transport function. All mutants were expressed at the plasma membrane. Biotinylation of free sulfhydryl groups using maleimide-PEG(11)-biotin and SDS-PAGE analysis under reducing and nonreducing conditions provided direct evidence for the existence of an essential disulfide bond between Cys-180 and Cys-329. This disulfide bridge is very likely involved in forming or stabilizing the substrate binding site. PMID- 19549784 TI - Importin-beta is a GDP-to-GTP exchange factor of Ran: implications for the mechanism of nuclear import. AB - Ran-GTP interacts strongly with importin-beta, and this interaction promotes the release of the importin-alpha-nuclear localization signal cargo from importin beta. Ran-GDP also interacts with importin-beta, but this interaction is 4 orders of magnitude weaker than the Ran-GTP.importin-beta interaction. Here we use the yeast complement of nuclear import proteins to show that the interaction between Ran-GDP and importin-beta promotes the dissociation of GDP from Ran. The release of GDP from the Ran-GDP-importin-beta complex stabilizes the complex, which cannot be dissociated by importin-alpha. Although Ran has a higher affinity for GDP compared with GTP, Ran in complex with importin-beta has a higher affinity for GTP. This feature is responsible for the generation of Ran-GTP from Ran-GDP by importin-beta. Ran-binding protein-1 (RanBP1) activates this reaction by forming a trimeric complex with Ran-GDP and importin-beta. Importin-alpha inhibits the GDP exchange reaction by sequestering importin-beta, whereas RanBP1 restores the GDP nucleotide exchange by importin-beta by forming a tetrameric complex with importin-beta, Ran, and importin-alpha. The exchange is also inhibited by nuclear-transport factor-2 (NTF2). We suggest a mechanism for nuclear import, additional to the established RCC1 (Ran-guanine exchange factor) dependent pathway that incorporates these results. PMID- 19549787 TI - Aortic aneurysm in MAGIC syndrome successfully managed with combined anti-TNF alpha and stent grafting. PMID- 19549788 TI - Efficacy of rituximab in refractory and relapsing myositis with anti-JO1 antibodies: a report of two cases. PMID- 19549789 TI - The transport of high amounts of vascular endothelial growth factor by blood platelets underlines their potential contribution in systemic sclerosis angiogenesis. AB - OBJECTIVES: Altered angiogenesis is a characteristic feature in SSc and remains ill-understood. VEGF is believed to play a central role. Serum VEGF is elevated in SSc patients but questions remain concerning the source of circulating VEGF. Here we investigated platelet activation and the role of platelets as a source of VEGF and other angiogenic mediators in this disease. METHODS: A cohort of 40 patients with SSc was included. Age- and sex-matched healthy subjects and subjects presenting a primary RP were included as controls. Platelets were isolated, activated with thrombin and the secretion of VEGF, platelet derived growth factor, homodimeric form BB (PDGF-BB), TGF-beta1 and angiopoietins-1 and 2 measured. Plasma concentrations of these mediators and the functionality of platelet-derived VEGF were also studied. Platelet activation was assayed by measuring plasma beta-thromboglobulin and expression of P-selectin on platelets. The effect of iloprost on VEGF secretion by platelets was studied. RESULTS: Platelets from SSc patients, in contrast to controls, secreted large amounts of VEGF when activated, but not PDGF-BB, TGF-beta1 or angiopoietins. Increased expression of membrane P-selectin confirmed platelet activation in the patients. Iloprost inhibited VEGF secretion by platelets both in vivo and in vitro, through inhibition of platelet activation. CONCLUSIONS: Platelets transport high levels of VEGF in SSc. They may contribute to circulating VEGF because of ongoing activation in the course of the disease. If activated at the contact of injured endothelium, platelets may be important in the altered angiogenesis associated with the disease through the secretion of high levels of VEGF. PMID- 19549790 TI - Catastrophic anti-phospholipid syndrome associated with Escherichia coli O157 infection. PMID- 19549791 TI - Functional analyses reveal the greater potency of preadipocytes compared with adipocytes as endothelial cell activator under normoxia, hypoxia, and TNFalpha exposure. AB - Obesity is associated with a state of chronic low-grade inflammation. Immune cells accumulate in white adipose tissue (WAT). The vascular endothelium plays an interactive role in these infiltration and inflammatory processes. Mature and hypertrophic adipocytes are considered as the major adipogenic cell type secreting proinflammatory cytokines in WAT. In contrast, the proinflammatory capacity of preadipocytes and their role in endothelial cell activation have been neglected so far. To gain new insights into this molecular and cellular cross talk, we examined the proinflammatory expression and secretion of normoxia, hypoxia, and TNFalpha-treated human preadipocytes and adipocytes (SGBS cells) and their impact on human microvascular endothelial cell (HMEC-1) function. In this study, stimulation of HMEC-1 with conditioned media (CM) from preadipocytes increased endothelial ICAM-1 expression and monocyte adhesion but not adipocyte CM. After hypoxia and TNFalpha stimulation of SGBS cells, adipocyte-CM induced and preadipocyte-CM enhanced the monocyte adhesion. Concordantly, the expression of proinflammatory adipokines was considerably higher in preadipocytes than in adipocytes. SGBS-CM upregulated the phosphorylation of three MAPK pathways, STAT1/3, and c-Jun in HMEC-1, whereas the NF-kappaB pathway was not affected. Inhibitor experiments showed that monocyte/endothelial cell-cell adhesion and endothelial ICAM-1 expression was JNK and JAK-1/STAT1/3 pathway dependent and revealed IL-6 as a major mediator in CM increasing monocyte/endothelial cell-cell adhesion via the STAT1/3 pathway. Our study shows that preadipocytes rather than adipocytes operate as potent activators of endothelial cells. This can be enhanced in preadipocytes and induced in adipocytes by TNFalpha and hypoxia in a manner similar to what may occur in WAT in the etiology of obesity. PMID- 19549792 TI - Detection of EETs and HETE-generating cytochrome P-450 enzymes and the effects of their metabolites on myometrial and vascular function. AB - Cytochrome P-450 (CYP450) enzymes of the CYP2 and -4 family in humans metabolize arachidonic acid to generate bioactive epoxyeicosatrienenoic acids (EETs) and hydroxyeicosatetrenoic acids (HETEs). We report significantly higher levels of CYP 2J2 protein expression following the onset of labor (n = 6, P < 0.05), implying increased EET-generating capacity within the uterus. Myometrial relaxation to 8,9-EET and 5,6-EET was observed, with the latter being inhibited by preincubation with 1 muM paxilline and is supported by whole cell recordings showing a modest effect of 5,6-EET on myometrial outward-current density (n = 4, P < 0.05). Only 5,6-EET of the EETs tested affected vascular reactivity (n = 6). Both 12- and 20-HETE (n = 5-6) caused vasoconstriction of partially depolarized blood vessels, with glibenclamide (n = 5) enhancing the effect of 12-HETE alone. Our findings signify a role for CYP2C9/19, -2J2, and -4A11/22 in late pregnancy, possibly related to the synthesis of lipid metabolites and downstream effects on vascular remodeling in the term pregnant uterus. The presence of CYP4A11/22 and their resultant procontractile metabolites could argue either a role in the control and initiation of labor and/or modification of the vascular delivery system to influence blood flow to the laboring uterus. The differential effects of the EETs and HETEs in the pregnant human uterus identify the CYP pathway as a novel modulator of myometrial and vascular physiology during late pregnancy. PMID- 19549793 TI - Hypergravity modulates vitamin D receptor target gene mRNA expression in mice. AB - The possibility of pathological calcium metabolism is a critical health concern introduced by long-term space travel. Because vitamin D plays an important role in calcium homeostasis, we evaluated the effects of hypergravity on the expression of genes involved in vitamin D and calcium metabolism in ICR mice. When exposed to 2G hypergravity for 2 days, the mRNA expression of renal 25 hydroxyvitamin D 24-hydroxylase (Cyp24a1) was increased and that of 25 hydroxyvitamin D 1alpha-hydroxylase (Cyp27b1) was decreased. Although hypergravity decreased food intake and increased the expression of starvation induced genes, the changes in Cyp24a1 and Cyp27b1 expression were not due to starvation, suggesting that hypergravity affects these genes directly. Hypergravity decreased plasma 1alpha,25-dihydroxyvitamin D(3) levels in ICR mice, suggesting a consequence of decreased Cyp27b1 and increased Cyp24a1 expression. Although 1alpha-hydroxyvitamin D(3) [1alpha(OH)D(3)] treatment induced the expression of vitamin D receptor (VDR) target genes in the kidney of 2G-exposed ICR mice to similar levels as controls, 1alpha(OH)D(3) increased the intestinal expression of Cyp24a1 in ICR mice. Hypergravity-dependent changes of Cyp24a1 and Cyp27b1 expression were diminished in mice exposed to hypergravity for 14 days, which may represent an adaptation to hypergravity stress. Hypergravity exposure also increased Cyp24a1 expression in the kidney of C57BL/6J mice. We examined the effects of hypergravity on VDR-null mice and found that renal Cyp27b1 expression in VDR-null mice was decreased by hypergravity while renal Cyp24a1 expression was not detected in VDR-null mice. Thus hypergravity modifies the expression of genes involved in vitamin D metabolism. PMID- 19549794 TI - Specific attenuation of protein kinase phosphorylation in muscle with a high mitochondrial content. AB - Acute contractile activity increases the activation of protein kinases involved in signal transduction. We hypothesized that the contractile activity-induced kinase phosphorylation would occur to a lesser degree in muscle with elevated mitochondrial content. We compared red and white sections of tibialis anterior (TA) muscle with two- to threefold differences in mitochondrial volume, and we increased the mitochondrial content in the TA muscle by 40% with unilateral chronic stimulation-induced contractile activity (10 Hz, 7 days, 3 h/day). Both the chronically stimulated and the contralateral control muscles were then acutely stimulated in situ for 15 min (10 Hz). We investigated 1) the total protein content and 2) the phosphorylation of kinases important for mitochondrial biogenesis in skeletal muscle, including AMPKalpha and p44, p42, and p38 MAPKs, as well as Akt by immunoblotting. In response to chronic stimulation, a selective upregulation of kinase protein content was observed, suggesting unique transcriptional/translational processing for these enzymes. Inverse relationships were observed between mitochondrial volume and 1) kinase protein content and 2) basal levels of kinase phosphorylation. In general, the kinase phosphorylation response to acute exercise depended, in part, on the oxidative capacity of the fiber type, evidenced by a greater absolute level of acute contractile activity induced kinase signaling in muscle with a lower mitochondrial volume. The attenuation of contraction-evoked kinase phosphorylation in muscle with high mitochondrial content suggests that these proteins may become less sensitive to upstream signaling and require greater stimulation for activation to propagate these adaptive cues downstream toward transcription or translation events. PMID- 19549795 TI - Augmenting frameworks for appraising the practices of community-based health interventions. AB - This paper aims at augmenting the frameworks proposed by Rifkin in 1996 to distinguish between target-oriented and empowerment approaches to participation in community-based health interventions. In her paper, Rifkin defined three criteria: who makes decisions on resource allocation, expected outcome and outcome assessment. We propose five additional criteria: the definition of community, the characteristics of the capacity-building process, the leadership characteristics, the documentation process, and ethical issues regarding participation. Derived from our analysis of a community-based project, the proposed criteria are discussed in the light of the principles of Popular Education and other literature on community participation. The augmented frameworks are intended to assist health professionals and planners interested in the empowerment approach of community participation to consciously sharpen their practice. PMID- 19549796 TI - Population pharmacokinetic analysis of linezolid in patients with infectious disease: application to lower body weight and elderly patients. AB - Linezolid (Zyvox), belonging to oxazolidinone antibiotics, is commonly used for the treatment of patients infected with methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci. Although linezolid has been approved worldwide, the Japanese pharmacokinetic (PK) profile has not been characterized in detail. The objective of this study is to develop a population PK model for linezolid that can be applied to a Japanese population. This population PK model was established based on the 1 Japanese phase III and 4 Caucasian phase II/III studies. A total of 2539 linezolid plasma concentration measurements from 455 patients, aged 18 to 98 years and body weight 30 to 190.5 kg, were used for the analysis. The data were analyzed using nonlinear mixed effects modeling. Body weight (BW), age, ethnicity, and gender were investigated as covariates. The final model was validated by the bootstrap technique. The PK profiles of linezolid were described with a 1-compartment PK model with first-order absorption and first-order elimination. In the final population PK model, BW and age were influential covariates on clearance, and the distribution volume was affected by BW. The present population PK model of linezolid described well the PK profiles in Japanese patients who have lower BW and are relatively older compared with those in the United States/European Union. PMID- 19549799 TI - 'How are you?': what do you mean? PMID- 19549797 TI - The interaction of a high-fat diet and regular moderate intensity exercise on intestinal polyp development in Apc Min/+ mice. AB - Diet and exercise are two environmental factors that can alter colon cancer risk. The purpose of this study was to determine if regular moderate-intensity treadmill exercise training could attenuate polyp formation in Apc(Min/+) mice fed the Western-style diet. Four-week-old male Apc(Min/+) mice (n = 12 per group) were assigned to AIN-76A Control, AIN-76A Exercise, Western Control, or Western Exercise treatment groups. Mice were weaned to these diets and either subjected to regular moderate-intensity treadmill exercise (18 m/min, 60 min/d, 6 d/wk) or remained sedentary for 6 weeks. Mice fed the Western-style diet consumed approximately 14% more calories and had 42% more epididymal fat compared with mice fed the AIN-76A diet. Exercise had no effect on fat pad mass with either diet treatment. Exercise reduced total intestinal polyp number by 50% and the number of large polyps (>1 mm diameter) by 67% in AIN-76A-fed mice. The Western style diet increased polyp number by 75% when compared with AIN-76A-fed mice, but exercise did not decrease polyp number or alter polyp size in mice fed the Western-style diet. Markers of systemic inflammation and immune system function were improved with exercise in mice fed the AIN-76A diet. Mice fed the Western style diet showed more inflammation and immunosuppression, which were not completely ameliorated by exercise. These data suggest that the induction of adiposity, inflammation, and immunosuppression by the Western-style diet may compromise the beneficial effect of moderate-intensity exercise on the intestinal polyp burden in Apc(Min/+) mice. PMID- 19549800 TI - The effect of television news items on intimate partner violence murders. AB - BACKGROUND: This study has been carried out in order to explore the effect of the growing appearance of IPV in television news items on deaths by this cause. METHODS: Ecological study based on 340 deaths by intimate partner violence (IPV) and 3733 television news items covering this topic on Spanish television channels (2003 and 2007). Logistic regression analysis was carried out. Dependent variable: the difference between the number of IPV deaths in the 7 days following a news item broadcast and the number of IPV deaths in the 7 days before a media item. This outcome variable was conceptualised as a binary variable: increase vs. no increase. INDEPENDENT VARIABLES: days with IPV news items-in general, concerning deaths, measures or other IPV crimes. Adjusted variables: day of the week, month, year and public holidays. RESULTS: A small copycat effect was observed in the IPV murder rate (RR = 1.32, CI(95%) = 1.07 - 1.62) for IPV related news item days compared with days without such news. This effect continued when information about deaths was broadcast (RR = 1.28, CI(95%) = 1.01 1.62). News items concerning measures (RR = 0.90, CI(95%) = 0.82 - 0.98) or other IPV crimes (0.84, CI(95%) = 0.82 - 0.98) were related to a lower possibility of an increase in deaths compared with days with death-related news. CONCLUSION: Given the results observed in the case of IPV-related news, there is an evident need to develop a journalistic style guide in order to determine what type of information is recommended due to the potential positive or negative effects. PMID- 19549801 TI - Is health recognized in the EU's policy process? An analysis of the European Commission's impact assessments. AB - BACKGROUND: The European Commission has an Impact Assessment (IA) procedure that aims to inform decision-makers of the all important impacts that decisions may have. This article studies how health is considered in the IA procedure and how it is reflected in the reports: what aspects, whose and simply in what context health is mentioned in the IA reports. METHODS: Half of the Commissions IAs from 2006 were studied. The analysis was text based and informed by content analysis. In total, 48 reports by 17 DGs were analysed. RESULTS: Five DGs (29%) and 10 reports (21%) made no reference to human health, public health or health systems. Five DGs were clearly considering health impacts more often than others; DG EMPL, SANCO, AGRI, ELARG and ENV. Health systems/services were most often and human health next most common referred to (39% and 29% of all, respectively). Health impacts were usually referred to in the sections on the definition of problems and the analysis of impacts. Seldom were they reported on in the sections on policy options, comparing options, or in the monitoring and evaluation sections. CONCLUSION: The results partly support concerns about the potential neglect of health impacts. The results also suggest that health is not considered an important factor when discussing alternative policy choices, and neither does it seem to be an important objective. There is a clear need for further exploration on ways in which health could be more appropriately considered when impacts of other policies are considered by the various DGs. PMID- 19549798 TI - Anti-inflammatory action of pterostilbene is mediated through the p38 mitogen activated protein kinase pathway in colon cancer cells. AB - Oxidative/nitrosative stress and generation of proinflammatory cytokines are hallmarks of inflammation. Because chronic inflammation is implicated in several pathologic conditions in humans, including cancers of the colon, anti inflammatory compounds may be useful chemopreventive agents against colon cancer. Stilbenes, such as resveratrol, have diverse pharmacologic activities, which include anti-inflammation, cancer prevention, a cholesterol-lowering effect, enhanced insulin sensitivity, and increased life span. We previously showed that pterostilbene (trans-3,5-dimethoxy-4'-hydroxystilbene), a structural analogue of resveratrol, is present in blueberries and that pterostilbene inhibited expression of certain inflammation-related genes in the colon and suppressed aberrant crypt foci formation in rats. Here, we examined molecular mechanisms of the action of pterostilbene in colon cancer. Pterostilbene reduced cell proliferation, down-regulated the expression of c-Myc and cyclin D1, and increased the level of cleaved poly(ADP-ribose) polymerase. A combination of cytokines (tumor necrosis factor-alpha, IFN-gamma, and bacterial endotoxin lipopolysaccharide) induced inflammation-related genes such as inducible nitric oxide synthase and cyclooxygenase-2, which was significantly suppressed by treatment with pterostilbene. We further identified upstream signaling pathways contributing to the anti-inflammatory activity of pterostilbene by investigating multiple signaling pathways, including nuclear factor-kappaB, Janus-activated kinase-signal transducer and activator of transcription, extracellular signal regulated kinase, p38, c-Jun NH(2)-terminal kinase, and phosphatidylinositol 3 kinase. Cytokine induction of the p38-activating transcription factor 2 pathway was markedly inhibited by pterostilbene among the different mediators of signaling evaluated. By silencing the expression of the p38 alpha isoform, there was significant reduction in cytokine induction of inducible nitric oxide synthase and cyclooxygenase-2. Our data suggest that the p38 mitogen-activated protein kinase cascade is a key signal transduction pathway for eliciting the anti-inflammatory action of pterostilbene in cultured HT-29 colon cancer cells. PMID- 19549802 TI - Educational inequalities in self-rated health within the Arab minority in Israel: explanatory factors. AB - BACKGROUND: Studies on factors explaining the socioeconomic inequalities in health within ethnic minorities are scarce. This study examines the contribution of material conditions, and psychosocial, community, cultural and behavioural factors to explaining educational disparities in self-rated health (SRH) within the Arab minority in Israel. METHODS: In 2005, a national random sample of 902 persons aged 30-70 was selected in a multistage sampling procedure for interviewing. We used multistage logistic regression modelling to examine the percent of attenuations in odds ratios (OR) of extreme categories for education (low vs. high) in the associations with SRH after inclusion of explanatory variables. RESULTS: Education was significantly associated with SRH [odds ratio (OR) = 3.86, 95% confidence interval (CI) = 2.30-6.47]. The separate inclusion of material conditions reduced the OR of poor SRH by 43%. Psychosocial, community or behavioural variables reduced it by 17%, 26% and 11%, respectively. Combining each group of factors with material conditions led to small additional reductions in OR. Integrating all explanatory variables reduced OR by 54%, with the association becoming non-significant. Cultural variables were not associated with SRH and were therefore not included in the multivariate analysis. CONCLUSIONS: Material conditions are a major factor in explaining the educational disparities in SRH among Arabs in Israel. Most of the impact of psychosocial and health behaviours, together with part of the community effects, operated through material conditions. Reduction of educational inequalities in SRH requires provision of educational attainment opportunities accompanied by employment opportunities assuring equivalent material gains. Further examination of factors explaining health inequalities among other minorities is warranted. PMID- 19549803 TI - Pneumonia mortality in a UK general practice population cohort. AB - BACKGROUND: Pneumonia is a common diagnosis in general practice in the United Kingdom and yet there is little known about the short- and long-term prognosis of people with a diagnosis of pneumonia in general practice. We investigated the short- and long-term survival of people with pneumonia diagnosed in general practice as compared to the general population for all ages. METHODS: This was a general population-based cohort study. Data was obtained from a comprehensive general practice database called The Health Improvement Network (THIN) database which has computerized medical records from 300 general practice surgeries in the United Kingdom. We used Cox regression for our analyses. RESULTS: For pneumonia cases the 30-day mortality was 18.5% and the 3-year mortality was 30.8%. The equivalent figures for the general population controls were 0.4% and 10.3% respectively. The adjusted hazard ratio (HR) for all-cause mortality (for total follow-up time) in pneumonia cases vs. general population was 4.64 (95% CI 4.35 4.95). For the first 30 days the risk of mortality in cases was 46 times more (adj. HR 45.90, 95% CI 36.80-55.20). Even in the period of follow-up 91 days after diagnosis cases were almost 20% more likely to die compared to general population (adj. HR 1.19, 95% CI 1.08-1.31). CONCLUSION: People in general practice who have a diagnosis of pneumonia have a markedly increased mortality in the short-term but some increase in mortality persists during longer-term follow up. PMID- 19549804 TI - Optimized detection of differential expression in global profiling experiments: case studies in clinical transcriptomic and quantitative proteomic datasets. AB - Identification of reliable molecular markers that show differential expression between distinct groups of samples has remained a fundamental research problem in many large-scale profiling studies, such as those based on DNA microarray or mass spectrometry technologies. Despite the availability of a wide spectrum of statistical procedures, the users of the high-throughput platforms are still facing the crucial challenge of deciding which test statistic is best adapted to the intrinsic properties of their own datasets. To meet this challenge, we recently introduced an adaptive procedure, named ROTS (Reproducibility-Optimized Test Statistic), which learns an optimal statistic directly from the given data, and whose relative benefits have previously been shown in comparison with state of-the-art procedures for detecting differential expression. Using gene expression microarray and mass-spectrometry (MS)-based protein expression datasets as case studies, we illustrate here the practical usage and advantages of ROTS toward detecting reliable marker lists in clinical transcriptomic and proteomic studies. In a public leukemia microarray dataset, the procedure could improve the sensitivity of the gene marker lists detected with high specificity. When applied to a recent LC-MS dataset, involving plasma samples from severe burn patients, the procedure could identify several peptide markers that remained undetected in the conventional analysis, thus demonstrating the effectiveness of ROTS also for global quantitative proteomic studies. To promote its widespread usage, we have made freely available efficient implementations of ROTS, which are easily accessible either as a stand-alone R-package or as integrated in the open source data analysis software Chipster. PMID- 19549805 TI - Increased activity of the oncogenic fatty acid synthase and the impaired glucose uptake in the metabolic syndrome. PMID- 19549806 TI - Latinas and breast cancer outcomes: population-based sampling, ethnic identity, and acculturation assessment. AB - PURPOSE: Latinas and African-Americans with breast cancer, especially those of lower socioeconomic status and acculturation, have been underrepresented in studies assessing treatment satisfaction, decision-making, and quality of life. A study was designed to recruit a large and representative sample of these subgroups. MATERIALS AND METHODS: Incident cases were selected by rapid case ascertainment (RCA) in the Los Angeles Surveillance, Epidemiology, and End Results Registry from 2005 to 2006, with oversampling of Latinas and African Americans. Patients were mailed a questionnaire and $10 incentive 5 to 6 months after diagnosis; nonrespondents were contacted by telephone. Multivariate analysis was used to assess possible response bias. The RCA definition of Hispanic origin was validated by self-reports. The Short Acculturation Scale for Hispanics index for Latina respondents was used. RESULTS: One thousand six hundred and ninety-eight eligible breast cancer cases were selected and 1,223 participated, for a response rate of 72.0%, which varied little by race/ethnicity. Age, race/ethnicity, and clinical factors were not associated with response; however, respondents were slightly more likely to be married and from higher socioeconomic status census tracts than nonrespondents. The RCA definition of Hispanic identity was highly sensitive (94.6%) and specific (90.0%). Lower acculturation was associated with lower education and literacy among Latinas. DISCUSSION: High response rates among all subgroups were achieved due to the use of RCA, an incentive, extensive telephone follow-up, a native Spanish-speaking interviewer, and a focused questionnaire. The low acculturation index category identified a highly vulnerable subgroup. This large sample representing subgroups with greater problems will provide a basis for developing better interventions to assist these women. PMID- 19549808 TI - Oral contraceptive use and BRCA penetrance: a case-only study. AB - BACKGROUND: Women with deleterious mutations in BRCA genes are at increased risk of breast cancer. However, the penetrance of the genetic trait may be regulated through environmental factors. This multinational case-only study tested the interaction between oral contraceptive use and genetic susceptibility in the occurrence of breast cancer. METHODS: We recruited 3,123 patients diagnosed with breast cancer before the age of 45 years. Participants were classified according to their probability of carrying a BRCA mutation on the basis of their family history of breast and ovarian cancer. According to a case-only approach, the frequency of relevant exposures among breast cancer cases with high probability of BRCA mutation ("genetic cases") was compared with the frequency of the same exposures among breast cancer cases with a low probability of BRCA mutation ("sporadic cases"). The interaction odds ratios (OR) and 95% confidence intervals (CI) for oral contraceptive use were estimated by unconditional logistic regression, after controlling for potentially confounding variables. RESULTS: The analysis was carried out comparing 382 "genetic" and 1,333 "sporadic" cases. We found a borderline significant interaction between genetic breast cancer and oral contraceptive use for ever users compared with never users (OR, 1.3; 95% CI, 1.0 1.7). The greatest interaction OR was found for women who started using pill at 18 to 20 years (OR, 1.6; 95% CI, 1.1-2.3). CONCLUSION: These results suggest that BRCA mutation carriers, as well as women with a significant family history of breast and ovarian cancer are more vulnerable to exogenous hormones in oral contraceptives. PMID- 19549807 TI - Prostate cancer risk associated loci in African Americans. AB - Four genome-wide association studies, all in populations of European descent, have identified 20 independent single nucleotide polymorphisms (SNP) in 20 regions that are associated with prostate cancer risk. We evaluated these 20 SNPs in a combined African American (AA) study, with 868 prostate cancer patients and 878 control subjects. For 17 of these 20 SNPs, implicated risk-associated alleles were found to be more common in these AA cases than controls, significantly more than expected under the null hypothesis (P = 0.03). Two of these 17 SNPs, located at 3p12, and region 2 at 8q24, were significantly associated with prostate cancer risk (P < 0.05), and only SNP rs16901979 at region 2 of 8q24 remained significant after accounting for 20 tests. A multivariate analysis of additional SNPs across the broader 8q24 region revealed three independent prostate cancer risk associated SNPs, including rs16901979, rs13254738, and rs10086908. The first two SNPs were approximately 20 kb apart and the last SNP, a novel finding from this study, was approximately 100 kb centromeric to the first two SNPs. These results suggest that a systematic evaluation of regions harboring known prostate cancer risk SNPs implicated in other races is an efficient approach to identify risk alleles for AA. However, studies with larger numbers of AA subjects are needed, and this will likely require a major collaborative effort to combine multiple AA study populations. PMID- 19549811 TI - Dietary intake of vegetables and fruits and the modification effects of GSTM1 and NAT2 genotypes on bladder cancer risk. AB - We analyzed the association between intakes of vegetables and fruits as defined by the U.S. Department of Agriculture pyramid food groups and bladder cancer risk using data collected in a large case-control study. The study included 884 histologically confirmed bladder cancer cases and 878 healthy controls matched to cases by age (+/-5 years), gender, and ethnicity. Significant inverse associations were observed for intakes of total vegetables, cruciferous vegetables, orange vegetables, dark green vegetables, and bladder cancer risk. Compared with those in the lowest quartile of total vegetable intake, the odds ratios for the 2nd, 3rd, and 4th quartiles of total vegetable intake were 0.84 [95% confidence interval (95% CI), 0.64-1.10], 0.71 (95% CI, 0.54-0.95), and 0.67 (95% CI, 0.50-0.90), respectively (P for trend = 0.004). Compared with those in the lowest quartile, those in the highest quartile of cruciferous vegetable intake had an odds ratio of 0.69 (95% CI, 0.52-0.92; P for trend = 0.001) and those in the highest quartile of orange vegetable intake had an odds ratio of 0.68 (95% CI, 0.52-0.91; P for trend = 0.006). Furthermore, the protective effect of cruciferous vegetables was more evident in subjects carrying GSTM1-null (odds ratio, 0.43; 95% CI, 0.25-0.73 for the 4th quartile of intake) and NAT2-slow genotypes (odds ratio, 0.56; 95% CI, 0.33-0.97 for the 4th quartile of intake). No association was observed for intakes of total fruits or citrus fruits. Our data strongly support that high vegetable consumption, especially cruciferous vegetable intake, may protect against bladder cancer and that genetic variants of GSTM1 and NAT2 may modify the association. PMID- 19549809 TI - Fine-mapping and family-based association analyses of prostate cancer risk variants at Xp11. AB - Two single nucleotide polymorphisms (SNP; rs5945572 and rs5945619) at Xp11 were recently implicated in two genome-wide association studies of prostate cancer. Using a family-based association test for these two SNPs in 168 families with prostate cancer, we showed in this study that the risk alleles of the two reported SNPs were overtransmitted to the affected offspring (P= 0.009 for rs5945372 and P = 0.03 for rs5945619), which suggested that the observed association in case-control studies were not driven by potential population stratification. We also did a fine-mapping study in the approximately 800 kb region at Xp11 between two independent case-control studies, including 1,527 cases and 482 controls from Johns Hopkins Hospital and 1,172 cases and 1,157 controls from the Prostate, Lung, Colon and Ovarian Cancer screening trial. The strongest association was found with SNPs in the haplotype block in which the two initial reported SNPs were located, although many SNPs in the approximately 140 kb region were highly significant in the combined allelic tests (P = 10(-5) to 10(-6)). The second strongest association was observed with SNPs in the approximately 286 kb region at another haplotype block (P = 10(-4) to 10(-5)), approximately 94 kb centromeric to the first region. The significance of SNPs in the second region decreased considerably after adjusting for SNPs at the first region, although P values remained at <0.05. Additional studies are warranted to test independent prostate cancer associations at these two regions. PMID- 19549810 TI - Meat and heterocyclic amine intake, smoking, NAT1 and NAT2 polymorphisms, and colorectal cancer risk in the multiethnic cohort study. AB - BACKGROUND: N-acetyltransferases (NAT) 1 and 2 are polymorphic enzymes catalyzing the metabolic activation of heterocyclic amines. We investigated the modifying effects of NAT1 and NAT2 polymorphisms on the association of meat consumption, heterocyclic amine intake, and smoking with colorectal cancer risk. METHOD: In the Multiethnic Cohort study, participants completed a smoking history and a food frequency questionnaire at recruitment and a cooked meat module 5 years later to estimate heterocyclic amine intake (2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, 2-amino-3,4,8 trimethylimidazo[4,5-f]quinoxaline). Blood samples were collected from incident cases and age-, sex-, ethnicity-, frequency-matched controls to determine genotypes. For analysis of meat intake and smoking, data were available for 1,009 cases and 1,522 controls; for heterocyclic amine intake analyses, 398 cases and 1,444 controls were available. Multivariate logistic regression models were used to estimate odds ratios. RESULTS: Smoking was associated with an increased colorectal cancer risk (odds ratio, 1.51; 95% confidence interval, 1.17-1.95) for > or =30 pack-years compared with never smokers (P trend = 0.0004). The association was stronger with presence of the "rapid" compared with the "slow/intermediate" NAT2 genotype (P interaction = 0.003). No significant associations were observed for intakes of red meat, processed meat, and heterocyclic amine, or meat doneness preference, but a dietary pattern high in meat showed a weak positive interaction with the NAT2 genotype (P interaction = 0.05). CONCLUSION: The enhanced association between smoking and colorectal cancer risk in subjects with the NAT2 rapid genotype supports a role for NAT2 and tobacco smoke heterocyclic amines in the etiology of colorectal cancer. This study only provides weak support for a similar association with meat heterocyclic amines. PMID- 19549812 TI - The impact of type 2 diabetes on the development of hepatocellular carcinoma in different viral hepatitis statuses. AB - BACKGROUND: The risk of type 2 diabetes on the development of hepatocellular carcinoma remains inconclusive in different hepatitis statuses. METHODS: We prospectively followed a community-based cohort with 5,929 persons in southern Taiwan from January 1997 through December 2004, made up of 4,117 seronegative, 982 anti-hepatitis C virus-positive [HCV(+)], 696 hepatitis B surface antigen positive [HBsAg(+)], and 134 coinfected persons. Before the study, 546 participants had developed diabetes. Hepatocellular carcinoma diagnoses were from the National Cancer Registry. RESULTS: After 50,899 person-years of follow-up, 111 individuals had developed hepatocellular carcinoma. The highest risk of hepatocellular carcinoma, compared with seronegative individuals without diabetes, was in anti-HCV(+) individuals with diabetes [incidence rate ratio (IRR), 76.0], then coinfected (IRR, 46.0), anti-HCV(+) without diabetes (IRR, 26.1), HBsAg(+) with diabetes (IRR, 21.4), and seronegative with diabetes (IRR, 7.2; P < 0.001). Anti-HCV(+) (n = 132) and seronegative individuals (n = 352) with diabetes had a higher cumulative incidence rate of hepatocellular carcinoma than those without diabetes (log-rank test, P < 0.001). Multivariate Cox proportional hazards analysis showed that gender, age, body mass index > or =30, HBsAg(+) [hazards ratio (HR), 12.6], anti-HCV(+) (HR, 18.8), coinfection (HR, 25.9), and diabetes [HR, 2.7; 95% confidence interval (95% CI), 1.7-4.3] were independent predictors of hepatocellular carcinoma (P < 0.05). After stratifying hepatitis status in multivariate Cox analysis, diabetes was significant for seronegative (HR, 5.4; 95% CI, 1.7-17.1) and anti-HCV(+) individuals (HR, 3.1; 95% CI, 1.7-5.4). Body mass index > or =30 was significant for HBsAg(+) individuals (HR, 3.3; 95% CI, 1.3-8.1). CONCLUSION: Type 2 diabetes is a strong independent predictor of hepatocellular carcinoma in anti-HCV(+) and seronegative individuals but not in HBsAg(+) individuals. PMID- 19549813 TI - Physiological and toxicological transcriptome changes in HepG2 cells exposed to copper. AB - Copper is an essential trace element; however, at supraphysiological levels, it can be extremely toxic. Microarray data from HepG2 cells exposed to 100, 200, 400, and 600 microM copper for 4, 8, 12 and 24 h were generated and analyzed. Principal components, K-means, and hierarchical clustering, interactome, and pathway mapping analyses indicated that these exposure conditions induce physiological and toxicological changes in the HepG2 transcriptome. As a general trend, when the level of toxicity increases, the number and diversity of affected genes, Gene Ontology categories, regulatory pathways, and complexity of interactomes increase. Physiological responses to copper include transition metal ion binding and responses to stress/stimulus, whereas toxicological responses include apoptosis, morphogenesis, and negative regulation of biomolecule metabolism. The global gene expression profile was overlaid onto biomolecular interaction networks and signal transduction cascades using pathway mapping and interactome identification. This analysis indicated that copper modulates signal transduction pathways associated with MAPK, NF-kappaB, death receptor, IGF-I, hypoxia, IL-10, IL-2, IL-6, EGF, Toll-like receptor, protein ubiquitination, xenobiotic metabolism, leukocyte extravasation, complement and coagulation, and sonic hedgehog signaling. These results provide insights into the global and molecular mechanisms regulating the physiological and toxicological responses to metal exposure. PMID- 19549815 TI - The dopamine puzzle. PMID- 19549816 TI - Tunable interplay between epidermal growth factor and cell-cell contact governs the spatial dynamics of epithelial growth. AB - Contact-inhibition of proliferation constrains epithelial tissue growth, and the loss of contact-inhibition is a hallmark of cancer cells. In most physiological scenarios, cell-cell contact inhibits proliferation in the presence of other growth-promoting cues, such as soluble growth factors (GFs). How cells quantitatively reconcile the opposing effects of cell-cell contact and GFs, such as epidermal growth factor (EGF), remains unclear. Here, using quantitative analysis of single cells within multicellular clusters, we show that contact is not a "master switch" that overrides EGF. Only when EGF recedes below a threshold level, contact inhibits proliferation, causing spatial patterns in cell cycle activity within epithelial cell clusters. Furthermore, we demonstrate that the onset of contact-inhibition and the timing of spatial patterns in proliferation may be reengineered. Using micropatterned surfaces to amplify cell-cell interactions, we induce contact-inhibition at a higher threshold level of EGF. Using a complementary molecular genetics approach to enhance cell-cell interactions by overexpressing E-cadherin also increases the threshold level of EGF at which contact-inhibition is triggered. These results lead us to propose a state diagram in which epithelial cells transition from a contact-uninhibited state to a contact-inhibited state at a critical threshold level of EGF, a property that may be tuned by modulating the extent of cell-cell contacts. This quantitative model of contact-inhibition has direct implications for how tissue size may be determined and deregulated during development and tumor formation, respectively, and provides design principles for engineering epithelial tissue growth in applications such as tissue engineering. PMID- 19549817 TI - The bactericidal effect of a complement-independent antibody is osmolytic and specific to Borrelia. AB - A complement-independent bactericidal IgG1 against the OspB of Borrelia burgdorferi increased the permeability of the outer membrane through the creation of openings of 2.8 - 4.4 nm, resulting in its osmotic lysis. Cryo-electron microscopy and tomography demonstrated that exposure to the antibody causes the formation of outer membrane projections and large breaks which may precede the increase in permeability of the outer membrane. The bactericidal effect of this antibody is not transferable to Escherichia coli expressing rOspB on its outer membrane. Additionally, the porin P66, the only protein that coprecipitated with OspB, is dispensable for the bactericidal mechanism. PMID- 19549819 TI - Protecting the Amazon with protected areas. AB - This article addresses climate-tipping points in the Amazon Basin resulting from deforestation. It applies a regional climate model to assess whether the system of protected areas in Brazil is able to avoid such tipping points, with massive conversion to semiarid vegetation, particularly along the south and southeastern margins of the basin. The regional climate model produces spatially distributed annual rainfall under a variety of external forcing conditions, assuming that all land outside protected areas is deforested. It translates these results into dry season impacts on resident ecosystems and shows that Amazonian dry ecosystems in the southern and southeastern basin do not desiccate appreciably and that extensive areas experience an increase in precipitation. Nor do the moist forests dry out to an excessive amount. Evidently, Brazilian environmental policy has created a sustainable core of protected areas in the Amazon that buffers against potential climate-tipping points and protects the drier ecosystems of the basin. Thus, all efforts should be made to manage them effectively. PMID- 19549818 TI - Membrane depolarization causes a direct activation of G protein-coupled receptors leading to local Ca2+ release in smooth muscle. AB - Membrane depolarization activates voltage-dependent Ca(2+) channels (VDCCs) inducing Ca(2+) release via ryanodine receptors (RyRs), which is obligatory for skeletal and cardiac muscle contraction and other physiological responses. However, depolarization-induced Ca(2+) release and its functional importance as well as underlying signaling mechanisms in smooth muscle cells (SMCs) are largely unknown. Here we report that membrane depolarization can induce RyR-mediated local Ca(2+) release, leading to a significant increase in the activity of Ca(2+) sparks and contraction in airway SMCs. The increased Ca(2+) sparks are independent of VDCCs and the associated extracellular Ca(2+) influx. This format of local Ca(2+) release results from a direct activation of G protein-coupled, M(3) muscarinic receptors in the absence of exogenous agonists, which causes activation of Gq proteins and phospholipase C, and generation of inositol 1,4,5 triphosphate (IP(3)), inducing initial Ca(2+) release through IP(3) receptors and then further Ca(2+) release via RyR2 due to a local Ca(2+)-induced Ca(2+) release process. These findings demonstrate an important mechanism for Ca(2+) signaling and attendant physiological function in SMCs. PMID- 19549814 TI - Stimulation of growth and changes in the hepatic transcriptome by 17beta estradiol in the yellow perch (Perca flavescens). AB - The effects of dietary 17beta-estradiol (E(2)) on growth and liver transcriptomics were investigated in the yellow perch (Perca flavescens). After a 3-mo treatment, E(2) significantly stimulated an increase in length and weight of juvenile male and female perch relative to control animals. The increase was significantly greater in females compared with males. Separate, unnormalized cDNA libraries were constructed from equal quantities of RNA from 6 male and 6 female livers of E(2)-treated and control perch, and 3,546 and 3,719 expressed sequence tags (ESTs) were obtained, respectively. To characterize E(2)-regulated transcripts, EST frequencies between libraries were calculated within contiguous sequences that were assembled from the combined ESTs of both libraries. Frequencies were also determined in EST transcript groupings produced by aligning all of the ESTs from both libraries at the nucleotide level. From these analyses, there were 28 annotated transcripts that were regulated by 75% between libraries and for which there were at least 5 ESTs of the same transcript between libraries. Regulation of a subset (14) of these transcripts was confirmed by quantitative reverse transcription-polymerase chain reaction (QPCR). Transcripts that were upregulated by E(2) included reproduction-related proteins, binding proteins, and proteases and protease inhibitors. While not part of the transcript frequency analysis, QPCR showed significant upregulation of estrogen receptor esr1 and of insulin-like growth factor I (IGF-I) in E(2) livers. E(2) downregulated transcripts represented a variety of functional categories including components of the respiratory chain, lipid transport and metabolism, glycolysis, amino acid and nitrogen metabolism, binding proteins, a hydrolytic enzyme, and a transcriptional regulator. In perch it appears that exogenous estrogen drastically shifts liver metabolism toward the production of lipoproteins and carbohydrate binding proteins, and that the growth-promoting action may involve an increase in hepatic IGF-I production. PMID- 19549820 TI - Opposing effects of SWI/SNF and Mi-2/NuRD chromatin remodeling complexes on epigenetic reprogramming by EBF and Pax5. AB - Transcriptionally silent genes are maintained in inaccessible chromatin. Accessibility of these genes requires their modification by chromatin remodeling complexes (CRCs), which are recruited to promoters by sequence-specific DNA binding proteins. Early B-cell factor (EBF), which is crucial for B-cell lineage specification, reprograms mb-1 (Ig-alpha) promoters by increasing chromatin accessibility and initiating the loss of DNA methylation. In turn, this facilitates promoter activation by Pax5. Here, we investigated the roles of ATP dependent CRCs in these mechanisms. Fusion of EBF and Pax5 with the ligand binding domain of ERalpha allowed for 4-hydroxytamoxifen-dependent, synergistic activation of mb-1 transcription in plasmacytoma cells. Knock-down of the SWI/SNF ATPases Brg1 and Brm inhibited transcriptional activation by EBF:ER and Pax5:ER. In contrast, knock-down of the Mi-2/NuRD complex subunit Mi-2beta greatly enhanced chromatin accessibility and mb-1 transcription in response to the activators. The reduction of Mi-2beta also propagated DNA demethylation in response to EBF:ER and Pax5:ER, resulting in fully unmethylated mb-1 promoters. In EBF- or EBF/Pax5-deficient fetal liver cells, both EBF and Pax5 were required for efficient demethylation of mb-1 promoters. Together, our data suggest that Mi 2/NuRD is important for the maintenance of hypermethylated chromatin in B cells. We conclude that SWI/SNF and Mi-2/NuRD function in opposition to enable or limit the reprogramming of genes by EBF and Pax5 during B-cell development. PMID- 19549821 TI - Decoupling of the brain's default mode network during deep sleep. AB - The recent discovery of a circuit of brain regions that is highly active in the absence of overt behavior has led to a quest for revealing the possible function of this so-called default-mode network (DMN). A very recent study, finding similarities in awake humans and anesthetized primates, has suggested that DMN activity might not simply reflect ongoing conscious mentation but rather a more general form of network dynamics typical of complex systems. Here, by performing functional MRI in humans, it is shown that a natural, sleep-induced reduction of consciousness is reflected in altered correlation between DMN network components, most notably a reduced involvement of frontal cortex. This suggests that DMN may play an important role in the sustenance of conscious awareness. PMID- 19549822 TI - Single homopolypeptide chains collapse into mechanically rigid conformations. AB - Huntington's disease is linked to the insertion of glutamine (Q) in the protein huntingtin, resulting in polyglutamine (polyQ) expansions that self-associate to form aggregates. While polyQ aggregation has been the subject of intense study, a correspondingly thorough understanding of individual polyQ chains is lacking. Here we demonstrate a single molecule force-clamp technique that directly probes the mechanical properties of single polyQ chains. We have made polyQ constructs of varying lengths that span the length range of normal and diseased polyQ expansions. Each polyQ construct is flanked by the I27 titin module, providing a clear mechanical fingerprint of the molecule being pulled. Remarkably, under the application of force, no extension is observed for any of the polyQ constructs. This is in direct contrast with the random coil protein PEVK of titin, which readily extends under force. Our measurements suggest that polyQ chains form mechanically stable collapsed structures. We test this hypothesis by disrupting polyQ chains with insertions of proline residues and find that their mechanical extensibility is sensitive to the position of the proline interruption. These experiments demonstrate that polyQ chains collapse to form a heterogeneous ensemble of conformations that are mechanically resilient. We further use a heat annealing molecular dynamics protocol to extensively search the conformation space and find that polyQ can exist in highly mechanically stable compact globular conformations. The mechanical rigidity of these collapsed structures may exceed the functional ability of eukaryotic proteasomes, resulting in the accumulation of undigested polyQ sequences in vivo. PMID- 19549823 TI - The structure of alanyl-tRNA synthetase with editing domain. AB - Alanyl-tRNA synthetase (AlaRS) catalyzes synthesis of Ala-tRNA(Ala) and hydrolysis of mis-acylated Ser- and Gly-tRNA(Ala) at 2 different catalytic sites. Here, we describe the monomer structures of C-terminal truncated archaeal AlaRS, with both activation and editing domains in the apo form, in complex with an Ala AMP analog, and in a high-resolution lysine-methylated form. The structures show docking of the editing domain to the activation domain opposite from the predicted tRNA-binding surface. Thus, the editing site is positioned >35 A from the activation site, prompting us to model 2 different tRNA complexes: one binding tRNA at the activation site, and the other binding tRNA at the editing site. Interestingly, a gel-shift assay also implies the presence of 2 types of tRNA complex with different mobility. These results suggest that tRNA translocation via a canonical CCA flipping is unlikely to occur in AlaRS. The structure also demonstrated the binding of zinc in the editing site, in which the specific coordination of zinc would be facilitated by a conserved GGQ motif, implying that the editing mechanism may not be the same as in ThrRS. As Asn-194 in eubacterial AlaRS important for Ser misactivation is replaced by Thr-213 in archaeal AlaRS, a different Ser accommodation mechanism is proposed. PMID- 19549824 TI - Secretion of amyloidogenic gelsolin progressively compromises protein homeostasis leading to the intracellular aggregation of proteins. AB - Familial amyloidosis of Finnish type (FAF) is a systemic amyloid disease associated with the deposition of proteolytic fragments of mutant (D187N/Y) plasma gelsolin. We report a mouse model of FAF featuring a muscle-specific promoter to drive D187N gelsolin synthesis. This model recapitulates the aberrant endoproteolytic cascade and the aging-associated extracellular amyloid deposition of FAF. Amyloidogenesis is observed only in tissues synthesizing human D187N gelsolin, despite the presence of full-length D187N gelsolin and its 68-kDa cleavage product in blood-demonstrating the importance of local synthesis in FAF. Loss of muscle strength was progressive in homozygous D187N gelsolin mice. The presence of misfolding-prone D187N gelsolin appears to exacerbate the age associated decline in cellular protein homeostasis (proteostasis), reflected by the intracellular deposition of numerous proteins, a characteristic of the most common degenerative muscle disease of aging humans, sporadic inclusion body myositis. PMID- 19549825 TI - A unique virus release mechanism in the Archaea. AB - Little is known about the infection cycles of viruses infecting cells from Archaea, the third domain of life. Here, we demonstrate that the virions of the archaeal Sulfolobus islandicus rod-shaped virus 2 (SIRV2) are released from the host cell through a mechanism, involving the formation of specific cellular structures. Large pyramidal virus-induced protrusions transect the cell envelope at several positions, rupturing the S-layer; they eventually open out, thus creating large apertures through which virions escape the cell. We also demonstrate that massive degradation of the host chromosomes occurs because of virus infection, and that virion assembly occurs in the cytoplasm. Furthermore, intracellular viral DNA is visualized by flow cytometry. The results show that SIRV2 is a lytic virus, and that the host cell dies as a consequence of elaborated mechanisms orchestrated by the virus. The generation of specific cellular structures for a distinct step of virus life cycle is known in eukaryal virus-host systems but is unprecedented in cells from other domains. PMID- 19549826 TI - Structure of a serine protease poised to resynthesize a peptide bond. AB - The serine proteases are among the most thoroughly studied enzymes, and numerous crystal structures representing the enzyme-substrate complex and intermediates in the hydrolysis reactions have been reported. Some aspects of the catalytic mechanism remain controversial, however, especially the role of conformational changes in the reaction. We describe here a high-resolution (1.46 A) crystal structure of a complex formed between a cleaved form of bovine pancreatic trypsin inhibitor (BPTI) and a catalytically inactive trypsin variant with the BPTI cleavage site ideally positioned in the active site for resynthesis of the peptide bond. This structure defines the positions of the newly generated amino and carboxyl groups following the 2 steps in the hydrolytic reaction. Comparison of this structure with those representing other intermediates in the reaction demonstrates that the residues of the catalytic triad are positioned to promote each step of both the forward and reverse reaction with remarkably little motion and with conservation of hydrogen-bonding interactions. The results also provide insights into the mechanism by which inhibitors like BPTI normally resist hydrolysis when bound to their target proteases. PMID- 19549827 TI - Enhanced humoral immune responses against T-independent antigens in Fc alpha/muR deficient mice. AB - IgM is an antibody class common to all vertebrates that plays a primary role in host defenses against infection. Binding of IgM with an antigen initiates the complement cascade, accelerating cellular and humoral immune responses. However, the functional role of the Fc receptor for IgM in such immune responses remains obscure. Here we show that mice deficient in Fc alpha/muR, an Fc receptor for IgM expressed on B cells and follicular dendritic cells (FDCs), have enhanced germinal center formation and affinity maturation and memory induction of IgG3(+) B cells after immunization with T-independent (TI) antigens. Moreover, Fc alpha/muR-deficient mice show prolonged antigen retention by marginal zone B (MZB) cells and FDCs. In vitro studies demonstrate that interaction of the IgM immune complex with Fc alpha/muR partly suppress TI antigen retention by MZB cells. We further show that downregulation of complement receptor (CR)1 and CR2 or complement deprivation by in vivo injection with anti-CR1/2 antibody or cobra venom factor attenuates antigen retention by MZB cells and germinal center formation after immunization with TI antigens in Fc alpha/muR(-/-) mice. Taken together, these results suggest that Fc alpha/muR negatively regulates TI antigen retention by MZB cells and FDCs, leading to suppression of humoral immune responses against T-independent antigens. PMID- 19549828 TI - Molecular recognition and self-assembly special feature: Assembly and organization processes in DNA-directed colloidal crystallization. AB - We present an analysis of the key steps involved in the DNA-directed assembly of nanoparticles into crystallites and polycrystalline aggregates. Additionally, the rate of crystal growth as a function of increased DNA linker length, solution temperature, and self-complementary versus non-self-complementary DNA linker strands (1- versus 2-component systems) has been studied. The data show that the crystals grow via a 3-step process: an initial "random binding" phase resulting in disordered DNA-AuNP aggregates, followed by localized reorganization and subsequent growth of crystalline domain size, where the resulting crystals are well-ordered at all subsequent stages of growth. PMID- 19549829 TI - The current refugial rainforests of Sundaland are unrepresentative of their biogeographic past and highly vulnerable to disturbance. AB - Understanding the historical dynamics of forest communities is a critical element for accurate prediction of their response to future change. Here, we examine evergreen rainforest distribution in the Sunda Shelf region at the last glacial maximum (LGM), using a spatially explicit model incorporating geographic, paleoclimatic, and geologic evidence. Results indicate that at the LGM, Sundaland rainforests covered a substantially larger area than currently present. Extrapolation of the model over the past million years demonstrates that the current "island archipelago" setting in Sundaland is extremely unusual given the majority of its history and the dramatic biogeographic transitions caused by global deglaciation were rapid and brief. Compared with dominant glacial conditions, lowland forests were probably reduced from approximately 1.3 to 0.8 x 10(6) km(2) while upland forests were probably reduced by half, from approximately 2.0 to 1.0 x 10(5) km(2). Coastal mangrove and swamp forests experienced the most dramatic change during deglaciations, going through a complete and major biogeographic relocation. The Sundaland forest dynamics of fragmentation and contraction and subsequent expansion, driven by glacial cycles, occur in the opposite phase as those in the northern hemisphere and equatorial Africa, indicating that Sundaland evergreen rainforest communities are currently in a refugial stage. Widespread human-mediated reduction and conversion of these forests in their refugial stage, when most species are passing through significant population bottlenecks, strongly emphasizes the urgency of conservation and management efforts. Further research into the natural process of fragmentation and contraction during deglaciation is necessary to understand the long-term effect of human activity on forest species. PMID- 19549831 TI - Formation of harmful algal blooms cannot be explained by allelopathic interactions. AB - Many planktonic microalgae produce a range of toxins and may form harmful algal blooms. One hypothesis is that some toxins are allelopathic, suppressing the growth of competitors, and it has been suggested that allelopathy may be one important mechanism causing algal blooms. In a metaanalysis of recent experimental work, we looked for evidence that allelopathy may explain the initiation of algal blooms. With few exceptions, allelopathic effects were only significant at very high cell densities typical of blooms. We conclude that there is no experimental support for allelopathy at prebloom densities, throwing doubts on allelopathy as a mechanism in bloom formation. Most studies tested allelopathy using cell-free manipulations. With simple models we show that cell-free manipulations may underestimate allelopathy at low cell densities if effects are transmitted during cell-cell interactions. However, we suggest that the evolution of allelopathy under field conditions may be unlikely even if based on cell-cell interactions. The spatial dispersion of cells in turbulent flow will make it difficult for an allelopathic cell to receive an exclusive benefit, and a dispersion model shows that dividing cells are rapidly separated constraining clone selection. Instead, we propose that reported allelopathic effects may be nonadaptive side effects of predator-prey or casual parasitic cell-cell interactions. PMID- 19549830 TI - A model of the cell-autonomous mammalian circadian clock. AB - Circadian timekeeping by intracellular molecular clocks is evident widely in prokaryotes and eukaryotes. The clockworks are driven by autoregulatory feedback loops that lead to oscillating levels of components whose maxima are in fixed phase relationships with one another. These phase relationships are the key metric characterizing the operation of the clocks. In this study, we built a mathematical model from the regulatory structure of the intracellular circadian clock in mice and identified its parameters using an iterative evolutionary strategy, with minimum cost achieved through conformance to phase separations seen in cell-autonomous oscillators. The model was evaluated against the experimentally observed cell-autonomous circadian phenotypes of gene knockouts, particularly retention of rhythmicity and changes in expression level of molecular clock components. These tests reveal excellent de novo predictive ability of the model. Furthermore, sensitivity analysis shows that these knockout phenotypes are robust to parameter perturbation. PMID- 19549832 TI - Tentacled snakes turn C-starts to their advantage and predict future prey behavior. AB - Fish are elusive prey with a short-latency escape behavior--the C-start- initiated to either the left or right by a "race" between 2 giant Mauthner neurons in the fish brainstem. Water disturbances usually excite the ipsilateral neuron, which massively excites contralateral motor neurons, resulting in a rapid turn away from striking predators. Here, it is reported that tentacled snakes (Erpeton tentaculatus) exploit this normally adaptive circuitry by feinting with their body, triggering the Mauthner cell that is furthest from their head milliseconds before a ballistic strike is initiated. As a result, fish that were oriented parallel to the long axis of the snake's head most often turned toward the approaching jaws, sometimes swimming directly into the snake's mouth. When strikes were instead directed at fish oriented at a right angle to the snake's head, snakes anticipated future fish behavior by striking to where fish would later be if they escaped from the snake's body feint, which fish usually did. The results provide an example of a rare predator taking advantage of a prey's normally adaptive escape circuitry and suggest that the snake's sensory-motor system is adapted to predict future behavior. PMID- 19549833 TI - Suppressor of gamma response 1 (SOG1) encodes a putative transcription factor governing multiple responses to DNA damage. AB - The Arabidopsis sog1-1 (suppressor of gamma response) mutant was originally isolated as a second-site suppressor of the radiosensitive phenotype of seeds defective in the repair endonuclease XPF. Here, we report that SOG1 encodes a putative transcription factor. This gene is a member of the NAC domain [petunia NAM (no apical meristem) and Arabidopsis ATAF1, 2 and CUC2] family (a family of proteins unique to land plants). Hundreds of genes are normally up-regulated in Arabidopsis within an hour of treatment with ionizing radiation; the induction of these genes requires the damage response protein kinase ATM, but not the related kinase ATR. Here, we find that SOG1 is also required for this transcriptional up regulation. In contrast, the SOG1-dependent checkpoint response observed in xpf mutant seeds requires ATR, but does not require ATM. Thus, phenotype of the sog1 1 mutant mimics aspects of the phenotypes of both atr and atm mutants in Arabidopsis, suggesting that SOG1 participates in pathways governed by both of these sensor kinases. We propose that, in plants, signals related to genomic stress are processed through a single, central transcription factor, SOG1. PMID- 19549834 TI - Tyrosine kinase nerve growth factor receptor switches from prosurvival to proapoptotic activity via Abeta-mediated phosphorylation. AB - The present study shows that increased Abeta production in hippocampal neurons, due to a failure of NGF signal, induces an unexpected phosphorylation of tyrosine kinase receptor A (TrkA), followed by activation of the phospholipase C gamma (PLCgamma) pathway and neuronal death. Such phosphorylation seems causally connected with 2 kinases known be involved in amyloidogenesis, Src and CDK5, and associated with alpha and gamma secretase-mediated p75 processing. Pharmacologic inhibition of TrkA phosphorylation and partial silencing of TrkA and/or p75 receptors prevent PLCgamma activation and protect neurons from death. Concomitantly with these events, TrkA, p75, Abeta peptides, and PS1 protein coimmunoprecipitate, suggesting their direct interplay in the subsequent onset of apoptotic death. Together, these findings depict a cellular mechanism whereby the same cellular transducing system may invert its intracellular message from trophic and antiapoptotic to a death signaling, which could also have relevance in the onset of Alzheimer's disease. PMID- 19549835 TI - Clathrin adaptor AP1B controls adenovirus infectivity of epithelial cells. AB - Adenoviruses invading the organism via normal digestive or respiratory routes require the Coxsackie-adenovirus receptor (CAR) to infect the epithelial barrier cells. Because CAR is a component of tight junctions and the basolateral membrane and is normally excluded from the apical membrane, most epithelia are resistant to adenoviruses. However, we discovered that a specialized epithelium, the retinal pigment epithelium (RPE), anomalously expressed CAR at the apical surface and was highly susceptible to adenovirus infection. These properties of RPE cells correlated with the absence of the epithelial-specific clathrin adaptor AP1B. Furthermore, knockdown of this basolateral sorting adaptor in adenovirus resistant MDCK cells promoted apical localization of CAR and increased dramatically Adenovirus infectivity. Targeting assays showed that AP1B is required for accurate basolateral recycling of CAR after internalization. AP1B knock down MDCK cells missorted CAR from recycling endosomes to the apical surface. In summary, we have characterized the cellular machinery responsible for normal sorting of an adenovirus receptor and illustrated how tissue-specific variations in such machinery result in drastic changes in tissue-susceptibility to adenoviruses. PMID- 19549836 TI - Molecular mechanism of membrane constriction and tubulation mediated by the F-BAR protein Pacsin/Syndapin. AB - Peripheral membrane proteins of the Bin/amphiphysin/Rvs (BAR) and Fer-CIP4 homology-BAR (F-BAR) family participate in cellular membrane trafficking and have been shown to generate membrane tubules. The degree of membrane bending appears to be encoded in the structure and immanent curvature of the particular protein domains, with BAR and F-BAR domains inducing high- and low-curvature tubules, respectively. In addition, oligomerization and the formation of ordered arrays influences tubule stabilization. Here, the F-BAR domain-containing protein Pacsin was found to possess a unique activity, creating small tubules and tubule constrictions, in addition to the wide tubules characteristic for this subfamily. Based on crystal structures of the F-BAR domain of Pacsin and mutagenesis studies, vesiculation could be linked to the presence of unique structural features distinguishing it from other F-BAR proteins. Tubulation was suppressed in the context of the full-length protein, suggesting that Pacsin is autoinhibited in solution. The regulated deformation of membranes and promotion of tubule constrictions by Pacsin suggests a more versatile function of these proteins in vesiculation and endocytosis beyond their role as scaffold proteins. PMID- 19549838 TI - Computing generalized Langevin equations and generalized Fokker-Planck equations. AB - The Mori-Zwanzig formalism is an effective tool to derive differential equations describing the evolution of a small number of resolved variables. In this paper we present its application to the derivation of generalized Langevin equations and generalized non-Markovian Fokker-Planck equations. We show how long time scales rates and metastable basins can be extracted from these equations. Numerical algorithms are proposed to discretize these equations. An important aspect is the numerical solution of the orthogonal dynamics equation which is a partial differential equation in a high dimensional space. We propose efficient numerical methods to solve this orthogonal dynamics equation. In addition, we present a projection formalism of the Mori-Zwanzig type that is applicable to discrete maps. Numerical applications are presented from the field of Hamiltonian systems. PMID- 19549837 TI - Baseline brain energy supports the state of consciousness. AB - An individual, human or animal, is defined to be in a conscious state empirically by the behavioral ability to respond meaningfully to stimuli, whereas the loss of consciousness is defined by unresponsiveness. PET measurements of glucose or oxygen consumption show a widespread approximately 45% reduction in cerebral energy consumption with anesthesia-induced loss of consciousness. Because baseline brain energy consumption has been shown by (13)C magnetic resonance spectroscopy to be almost exclusively dedicated to neuronal signaling, we propose that the high level of brain energy is a necessary property of the conscious state. Two additional neuronal properties of the conscious state change with anesthesia. The delocalized fMRI activity patterns in rat brain during sensory stimulation at a higher energy state (close to the awake) collapse to a contralateral somatosensory response at lower energy state (deep anesthesia). Firing rates of an ensemble of neurons in the rat somatosensory cortex shift from the gamma-band range (20-40 Hz) at higher energy state to <10 Hz at lower energy state. With the conscious state defined by the individual's behavior and maintained by high cerebral energy, measurable properties of that state are the widespread fMRI patterns and high frequency neuronal activity, both of which support the extensive interregional communication characteristic of consciousness. This usage of high brain energies when the person is in the "state" of consciousness differs from most studies, which attend the smaller energy increments observed during the stimulations that form the "contents" of that state. PMID- 19549839 TI - Exocyst is involved in polarized cell migration and cerebral cortical development. AB - Neuronal migration is essential for proper development of the cerebral cortex. As a first step, a postmitotic cell extends its leading process, presumably by adding new membrane at the growing tip, which would enable directed locomotion. The goal of the present study was to determine if biosynthetic exocytic pathway is polarized in migrating cells and whether polarized exocytosis promotes directed cell migration. A promising candidate for controlling the spatial sites of vesicle tethering and fusion at the plasma membrane is a protein complex called the exocyst. We found that cell migration in a wound assay, as well as cortical neuronal migration during embryonic development was impaired when the exocyst was disturbed. By combining TIRF microscopy and a stochastic model of exocytosis, we found that vesicle exocytosis is preferentially distributed close to the leading edge of polarized cells, that the exocytic process is organized into hotspots, and that the polarized delivery of vesicles and their clustering in hotspots depend on the intact exocyst complex. The exocyst complex seems to achieve this spatial regulation by determining the sites at the membrane where secretory vesicles tether. Thus, our study supports the notion that polarized membrane traffic regulated by the exocyst is an essential component of cell migration and that its deficit may lead to cortical abnormalities involving cortical neuronal malpositioning. PMID- 19549841 TI - A two-stage algorithm for extracting the multiscale backbone of complex weighted networks. PMID- 19549840 TI - Selective induction of neocortical GABAergic neurons by the PDK1-Akt pathway through activation of Mash1. AB - Extracellular stimuli regulate neuronal differentiation and subtype specification during brain development, although the intracellular signaling pathways that mediate these processes remain largely unclear. We now show that the PDK1-Akt pathway regulates differentiation of telencephalic neural precursor cells (NPCs). Active Akt promotes differentiation of NPC into gamma-aminobutyric acid containing (GABAergic) but not glutamatergic neurons. Disruption of the Pdk1 gene or expression of dominant-negative forms of Akt suppresses insulin-like growth factor (IGF)-1 enhancement of NPC differentiation into neurons in vitro and production of neocortical GABAergic neurons in vivo. Furthermore, active Akt increased the protein levels and transactivation activity of Mash1, a proneural basic helix-loop-helix protein required for the generation of neocortical GABAergic neurons, and Mash1 was required for Akt-induced neuronal differentiation. These results have unveiled an unexpected role of the PDK1-Akt pathway: a key mediator of extracellular signals regulating the production of neocortical GABAergic neurons. PMID- 19549842 TI - Widespread metabolic potential for nitrite and nitrate assimilation among Prochlorococcus ecotypes. AB - The marine cyanobacterium Prochlorococcus is the most abundant photosynthetic organism in oligotrophic regions of the oceans. The inability to assimilate nitrate is considered an important factor underlying the distribution of Prochlorococcus, and thought to explain, in part, low abundance of Prochlorococcus in coastal, temperate, and upwelling zones. Here, we describe the widespread occurrence of a genomic island containing nitrite and nitrate assimilation genes in uncultured Prochlorococcus cells from marine surface waters. These genes are characterized by low GC content, form a separate phylogenetic clade most closely related to marine Synechococcus, and are located in a different genomic region compared with an orthologous cluster found in marine Synechococcus strains. This sequence distinction suggests that these genes were not transferred recently from Synechococcus. We demonstrate that the nitrogen assimilation genes encode functional proteins and are expressed in the ocean. Also, we find that their relative occurrence is higher in the Caribbean Sea and Indian Ocean compared with the Sargasso Sea and Eastern Pacific Ocean, which may be related to the nitrogen availability in each region. Our data suggest that the ability to assimilate nitrite and nitrate is associated with microdiverse lineages within high- and low-light (LL) adapted Prochlorococcus ecotypes. It challenges 2 long-held assumptions that (i) Prochlorococcus cannot assimilate nitrate, and (ii) only LL adapted ecotypes can use nitrite. The potential for previously unrecognized productivity by Prochlorococcus in the presence of oxidized nitrogen species has implications for understanding the biogeography of Prochlorococcus and its role in the oceanic carbon and nitrogen cycles. PMID- 19549843 TI - Increased InsP3Rs in the junctional sarcoplasmic reticulum augment Ca2+ transients and arrhythmias associated with cardiac hypertrophy. AB - Cardiac hypertrophy is a growth response of the heart to increased hemodynamic demand or damage. Accompanying this heart enlargement is a remodeling of Ca(2+) signaling. Due to its fundamental role in controlling cardiomyocyte contraction during every heartbeat, modifications in Ca(2+) fluxes significantly impact on cardiac output and facilitate the development of arrhythmias. Using cardiomyocytes from spontaneously hypertensive rats (SHRs), we demonstrate that an increase in Ca(2+) release through inositol 1,4,5-trisphosphate receptors (InsP(3)Rs) contributes to the larger excitation contraction coupling (ECC) mediated Ca(2+) transients characteristic of hypertrophic myocytes and underlies the more potent enhancement of ECC-mediated Ca(2+) transients and contraction elicited by InsP(3) or endothelin-1 (ET-1). Responsible for this is an increase in InsP(3)R expression in the junctional sarcoplasmic reticulum. Due to their close proximity to ryanodine receptors (RyRs) in this region, enhanced Ca(2+) release through InsP(3)Rs served to sensitize RyRs, thereby increasing diastolic Ca(2+) levels, the incidence of extra-systolic Ca(2+) transients, and the induction of ECC-mediated Ca(2+) elevations. Unlike the increase in InsP(3)R expression and Ca(2+) transient amplitude in the cytosol, InsP(3)R expression and ECC-mediated Ca(2+) transients in the nucleus were not altered during hypertrophy. Elevated InsP(3)R2 expression was also detected in hearts from human patients with heart failure after ischemic dilated cardiomyopathy, as well as in aortic-banded hypertrophic mouse hearts. Our data establish that increased InsP(3)R expression is a general mechanism that underlies remodeling of Ca(2+) signaling during heart disease, and in particular, in triggering ventricular arrhythmia during hypertrophy. PMID- 19549844 TI - BCL6 suppression of BCL2 via Miz1 and its disruption in diffuse large B cell lymphoma. AB - The BCL6 proto-oncogene encodes a transcriptional repressor that is required for germinal center (GC) formation and whose deregulation by genomic lesions is implicated in the pathogenesis of GC-derived diffuse large B cell lymphoma (DLBCL) and, less frequently, follicular lymphoma (FL). The biological function of BCL6 is only partially understood because no more than a few genes have been functionally characterized as direct targets of BCL6 transrepression activity. Here we report that the anti-apoptotic proto-oncogene BCL2 is a direct target of BCL6 in GC B cells. BCL6 binds to the BCL2 promoter region by interacting with the transcriptional activator Miz1 and suppresses Miz1-induced activation of BCL2 expression. BCL6-mediated suppression of BCL2 is lost in FL and DLBCL, where the 2 proteins are pathologically coexpressed, because of BCL2 chromosomal translocations and other mechanisms, including Miz1 deregulation and somatic mutations in the BCL2 promoter region. These results identify an important function for BCL6 in facilitating apoptosis of GC B cells via suppression of BCL2, and suggest that blocking this pathway is critical for lymphomagenesis. PMID- 19549845 TI - Oxidoreductase activity of oligosaccharyltransferase subunits Ost3p and Ost6p defines site-specific glycosylation efficiency. AB - Asparagine-linked glycosylation is a common posttranslational modification of diverse secretory and membrane proteins in eukaryotes, where it is catalyzed by the multiprotein complex oligosaccharyltransferase. The functions of the protein subunits of oligoasccharyltransferase, apart from the catalytic Stt3p, are ill defined. Here we describe functional and structural investigations of the Ost3/6p components of the yeast enzyme. Genetic, biochemical and structural analyses of the lumenal domain of Ost6p revealed oxidoreductase activity mediated by a thioredoxin-like fold with a distinctive active-site loop that changed conformation with redox state. We found that mutation of the active-site cysteine residues of Ost6p and its paralogue Ost3p affected the glycosylation efficiency of a subset of glycosylation sites. Our results show that eukaryotic oligosaccharyltransferase is a multifunctional enzyme that acts at the crossroads of protein modification and protein folding. PMID- 19549846 TI - Detection and differentiation of normal, cancerous, and metastatic cells using nanoparticle-polymer sensor arrays. AB - Rapid and effective differentiation between normal and cancer cells is an important challenge for the diagnosis and treatment of tumors. Here, we describe an array-based system for identification of normal and cancer cells based on a "chemical nose/tongue" approach that exploits subtle changes in the physicochemical nature of different cell surfaces. Their differential interactions with functionalized nanoparticles are transduced through displacement of a multivalent polymer fluorophore that is quenched when bound to the particle and fluorescent after release. Using this sensing strategy we can rapidly (minutes/seconds) and effectively distinguish (i) different cell types; (ii) normal, cancerous and metastatic human breast cells; and (iii) isogenic normal, cancerous and metastatic murine epithelial cell lines. PMID- 19549847 TI - Stoichiometric and temporal requirements of Oct4, Sox2, Klf4, and c-Myc expression for efficient human iPSC induction and differentiation. AB - Human-induced pluripotent stem cells (hiPSCs) are generated from somatic cells by ectopic expression of the 4 reprogramming factors (RFs) Oct-4, Sox2, Klf4, and c Myc. To better define the stoichiometric requirements and dynamic expression patterns required for successful hiPSC induction, we generated 4 bicistronic lentiviral vectors encoding the 4 RFs co-expressed with discernable fluorescent proteins. Using this system, we define the optimal stoichiometry of RF expression to be highly sensitive to Oct4 dosage, and we demonstrate the impact that variations in the relative ratios of RF expression exert on the efficiency of hiPSC induction. Monitoring of expression of each individual RF in single cells during the course of reprogramming revealed that vector silencing follows acquisition of pluripotent cell markers. Pronounced lentiviral vector silencing was a characteristic of successfully reprogrammed hiPSC clones, but lack of complete silencing did not hinder hiPSC induction, maintenance, or directed differentiation. The vector system described here presents a powerful tool for mechanistic studies of reprogramming and the optimization of hiPSC generation. PMID- 19549848 TI - Chromosomal translocations induced at specified loci in human stem cells. AB - The precise genetic manipulation of stem and precursor cells offers extraordinary potential for the analysis, prevention, and treatment of human malignancies. Chromosomal translocations are hallmarks of several tumor types where they are thought to have arisen in stem or precursor cells. Although approaches exist to study factors involved in translocation formation in mouse cells, approaches in human cells have been lacking, especially in relevant cell types. The technology of zinc finger nucleases (ZFNs) allows DNA double-strand breaks (DSBs) to be introduced into specified chromosomal loci. We harnessed this technology to induce chromosomal translocations in human cells by generating concurrent DSBs at 2 endogenous loci, the PPP1R12C/p84 gene on chromosome 19 and the IL2Rgamma gene on the X chromosome. Translocation breakpoint junctions for t(19;X) were detected with nested quantitative PCR in a high throughput 96-well format using denaturation curves and DNA sequencing in a variety of human cell types, including embryonic stem (hES) cells and hES cell-derived mesenchymal precursor cells. Although readily detected, translocations were less frequent than repair of a single DSB by gene targeting or nonhomologous end-joining, neither of which leads to gross chromosomal rearrangements. While previous studies have relied on laborious genetic modification of cells and extensive growth in culture, the approach described in this report is readily applicable to primary human cells, including multipotent and pluripotent cells, to uncover both the underlying mechanisms and phenotypic consequences of targeted translocations and other genomic rearrangements. PMID- 19549849 TI - The neural bases of empathic accuracy. AB - Theories of empathy suggest that an accurate understanding of another's emotions should depend on affective, motor, and/or higher cognitive brain regions, but until recently no experimental method has been available to directly test these possibilities. Here, we present a functional imaging paradigm that allowed us to address this issue. We found that empathically accurate, as compared with inaccurate, judgments depended on (i) structures within the human mirror neuron system thought to be involved in shared sensorimotor representations, and (ii) regions implicated in mental state attribution, the superior temporal sulcus and medial prefrontal cortex. These data demonstrate that activity in these 2 sets of brain regions tracks with the accuracy of attributions made about another's internal emotional state. Taken together, these results provide both an experimental approach and theoretical insights for studying empathy and its dysfunction. PMID- 19549850 TI - Essential role of the Ly49A stalk region for immunological synapse formation and signaling. AB - NK cells use surface NK receptors to discriminate self from non-self. The NK receptor ligand-binding domain (NKD) has been considered the sole regulator of ligand binding. Using a prototypic murine NK receptor, Ly49A, we show that the membrane proximal nonligand binding ecto-domain (the stalk region) is critical to ligand binding and signaling. The stalk region is required for receptor binding to ligand on target cells (trans interaction), but is dispensable for receptor binding to ligand on the same cell (cis interaction). Also, signaling in a trans manner depends on the stalk region mediating the formation of the immunological synapse. Thus, our data modeling receptor function at the cellular level reveal an essential role for the stalk region as a specific mediator of receptor signal integration, by which NKD-ligand interactions at the interface initiate and deliver information to the spatially separated cytoplasmic domain. PMID- 19549851 TI - A multiscale model linking ion-channel molecular dynamics and electrostatics to the cardiac action potential. AB - Ion-channel function is determined by its gating movement. Yet, molecular dynamics and electrophysiological simulations were never combined to link molecular structure to function. We performed multiscale molecular dynamics and continuum electrostatics calculations to simulate a cardiac K(+) channel (I(Ks)) gating and its alteration by mutations that cause arrhythmias and sudden death. An all-atom model of the I(Ks) alpha-subunit KCNQ1, based on the recent Kv1.2 structure, is used to calculate electrostatic energies during gating. Simulations are compared with experiments where varying degrees of positive charge-added via point mutation-progressively reduce current. Whole-cell simulations show that mutations cause action potential and ECG QT interval prolongation, consistent with clinical phenotypes. This framework allows integration of multiscale observations to study the molecular basis of excitation and its alteration by disease. PMID- 19549852 TI - NIH funding trajectories and their correlations with US health dynamics from 1950 to 2004. AB - To determine optimal future National Institutes of Health (NIH) funding levels, the longitudinal correlation of the level of investment in NIH research with population changes in the risk of specific diseases should be analyzed. This is because NIH research is the primary source of new therapies and treatments for major chronic diseases, many of which were viewed as relatively untreatable in the 1950s. NIH research is also important in developing preventative and screening strategies to support public health interventions. These correlations are examined 1938 to 2004 for 4 major chronic diseases [cardiovascular disease (CVD), stroke, cancer, and diabetes] and the NIH institutes responsible for research for those diseases. This analysis shows consistent non-linear temporal correlations of funding to mortality rates across diseases. The economic implications of this are discussed assuming that improved health at later ages will allow projected declines in the rate of growth of the US labor force to be partly offset by a higher rate of labor force participation in the US elderly population due to reduced chronic disease risks and functional impairment. PMID- 19549853 TI - SirT1 knockdown in liver decreases basal hepatic glucose production and increases hepatic insulin responsiveness in diabetic rats. AB - Hepatic gluconeogenesis is a major contributing factor to hyperglycemia in the fasting and postprandial states in type 2 diabetes mellitus (T2DM). Because Sirtuin 1 (SirT1) induces hepatic gluconeogenesis during fasting through the induction of phosphoenolpyruvate carboxylase kinase (PEPCK), fructose-1,6 bisphosphatase (FBPase), and glucose-6-phosphatase (G6Pase) gene transcription, we hypothesized that reducing SirT1, by using an antisense oligonucleotide (ASO), would decrease fasting hyperglycemia in a rat model of T2DM. SirT1 ASO lowered both fasting glucose concentration and hepatic glucose production in the T2DM rat model. Whole body insulin sensitivity was also increased in the SirT1 ASO treated rats as reflected by a 25% increase in the glucose infusion rate required to maintain euglycemia during the hyperinsulinemic-euglycemic clamp and could entirely be attributed to increased suppression of hepatic glucose production by insulin. The reduction in basal and clamped rates of glucose production could in turn be attributed to decreased expression of PEPCK, FBPase, and G6Pase due to increased acetylation of signal transducer and activator of transcription 3 (STAT3), forkhead box O1 (FOXO1), and peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha), known substrates of SirT1. In addition to the effects on glucose metabolism, SirT1 ASO decreased plasma total cholesterol, which was attributed to increased cholesterol uptake and export from the liver. These results indicate that inhibition of hepatic SirT1 may be an attractive approach for treatment of T2DM. PMID- 19549854 TI - Identification of a consensus motif in substrates bound by a Type I Hsp40. AB - Protein aggregation is a hallmark of a large and diverse number of conformational diseases. Molecular chaperones of the Hsp40 family (Escherichia coli DnaJ homologs) recognize misfolded disease proteins and suppress the accumulation of toxic protein species. Type I Hsp40s are very potent at suppressing protein aggregation and facilitating the refolding of damaged proteins. Yet, the molecular mechanism for the recognition of nonnative polypeptides by Type I Hsp40s such as yeast Ydj1 is not clear. Here we computationally identify a unique motif that is selectively recognized by Ydj1p. The motif is characterized by the consensus sequence GX[LMQ]{P}X{P}{CIMPVW}, where [XY] denotes either X or Y and {XY} denotes neither X nor Y. We further verify the validity of the motif by site directed mutagenesis and show that substrate binding by Ydj1 requires recognition of this motif. A yeast proteome screen revealed that many proteins contain more than one stretch of residues that contain the motif and are separated by varying numbers of amino acids. In light of our results, we propose a 2-site peptide binding model and a plausible mechanism of peptide presentation by Ydj1p to the chaperones of the Hsp70 family. Based on our results, and given that Ydj1p and its human ortholog Hdj2 are functionally interchangeable, we hypothesize that our results can be extended to understanding human diseases. PMID- 19549855 TI - Eukaryotic cytosolic and mitochondrial phenylalanyl-tRNA synthetases catalyze the charging of tRNA with the meta-tyrosine. AB - The accumulation of proteins damaged by reactive oxygen species (ROS), conventionally regarded as having pathological potentials, is associated with age related diseases such as Alzheimer's, atherosclerosis, and cataractogenesis. Exposure of the aromatic amino acid phenylalanine to ROS-generating systems produces multiple isomers of tyrosine: m-tyrosine (m-Tyr), o-tyrosine (o-Tyr), and the standard p-tyrosine (Tyr). Previously it was demonstrated that exogenously supplied, oxidized amino acids could be incorporated into bacterial and eukaryotic proteins. It is, therefore, likely that in many cases, in vivo damaged amino acids are available for de novo synthesis of proteins. Although the involvement of aminoacyl-tRNA synthetases in this process has been hypothesized, the specific pathway by which ROS-damaged amino acids are incorporated into proteins remains unclear. We provide herein evidence that mitochondrial and cytoplasmic phenylalanyl-tRNA synthetases (HsmtPheRS and HsctPheRS, respectively) catalyze direct attachment of m-Tyr to tRNA(Phe), thereby opening the way for delivery of the misacylated tRNA to the ribosome and incorporation of ROS-damaged amino acid into eukaryotic proteins. Crystal complexes of mitochondrial and bacterial PheRSs with m-Tyr reveal the net of highly specific interactions within the synthetic and editing sites. PMID- 19549856 TI - An integrated genome screen identifies the Wnt signaling pathway as a major target of WT1. AB - WT1, a critical regulator of kidney development, is a tumor suppressor for nephroblastoma but in some contexts functions as an oncogene. A limited number of direct transcriptional targets of WT1 have been identified to explain its complex roles in tumorigenesis and organogenesis. In this study we performed genome-wide screening for direct WT1 targets, using a combination of ChIP-ChIP and expression arrays. Promoter regions bound by WT1 were highly G-rich and resembled the sites for a number of other widely expressed transcription factors such as SP1, MAZ, and ZNF219. Genes directly regulated by WT1 were implicated in MAPK signaling, axon guidance, and Wnt pathways. Among directly bound and regulated genes by WT1, nine were identified in the Wnt signaling pathway, suggesting that WT1 modulates a subset of Wnt components and responsive genes by direct binding. To prove the biological importance of the interplay between WT1 and Wnt signaling, we showed that WT1 blocked the ability of Wnt8 to induce a secondary body axis during Xenopus embryonic development. WT1 inhibited TCF-mediated transcription activated by Wnt ligand, wild type and mutant, stabilized beta-catenin by preventing TCF4 loading onto a promoter. This was neither due to direct binding of WT1 to the TCF binding site nor to interaction between WT1 and TCF4, but by competition of WT1 and TCF4 for CBP. WT1 interference with Wnt signaling represents an important mode of its action relevant to the suppression of tumor growth and guidance of development. PMID- 19549858 TI - Evidence for electronic gap-driven metal-semiconductor transition in phase-change materials. AB - Phase-change materials are functionally important materials that can be thermally interconverted between metallic (crystalline) and semiconducting (amorphous) phases on a very short time scale. Although the interconversion appears to involve a change in local atomic coordination numbers, the electronic basis for this process is still unclear. Here, we demonstrate that in a nearly vacancy-free binary GeSb system where we can drive the phase change both thermally and, as we discover, by pressure, the transformation into the amorphous phase is electronic in origin. Correlations between conductivity, total system energy, and local atomic coordination revealed by experiments and long time ab initio simulations show that the structural reorganization into the amorphous state is driven by opening of an energy gap in the electronic density of states. The electronic driving force behind the phase change has the potential to change the interconversion paradigm in this material class. PMID- 19549857 TI - Controlling transgene expression in subcutaneous implants using a skin lotion containing the apple metabolite phloretin. AB - Adjustable control of therapeutic transgenes in engineered cell implants after transdermal and topical delivery of nontoxic trigger molecules would increase convenience, patient compliance, and elimination of hepatic first-pass effect in future therapies. Pseudomonas putida DOT-T1E has evolved the flavonoid-triggered TtgR operon, which controls expression of a multisubstrate-specific efflux pump (TtgABC) to resist plant-derived defense metabolites in its rhizosphere habitat. Taking advantage of the TtgR operon, we have engineered a hybrid P. putida mammalian genetic unit responsive to phloretin. This flavonoid is contained in apples, and, as such, or as dietary supplement, regularly consumed by humans. The engineered mammalian phloretin-adjustable control element (PEACE) enabled adjustable and reversible transgene expression in different mammalian cell lines and primary cells. Due to the short half-life of phloretin in culture, PEACE could also be used to program expression of difficult-to-produce protein therapeutics during standard bioreactor operation. When formulated in skin lotions and applied to the skin of mice harboring transgenic cell implants, phloretin was able to fine-tune target genes and adjust heterologous protein levels in the bloodstream of treated mice. PEACE-controlled target gene expression could foster advances in biopharmaceutical manufacturing as well as gene- and cell-based therapies. PMID- 19549859 TI - Amino acid polymorphisms altering the glycosylation of IL-2 do not protect from type 1 diabetes in the NOD mouse. AB - Idd3 is one of many gene regions that affect the development of type 1 diabetes (T1D) in the nonobese diabetic (NOD) mouse. Idd3 has been localized to a 650-kb region on chromosome 3 containing the IL-2 gene. Exon 1 of the IL-2 gene is polymorphic between the susceptible NOD and the protective C57BL/6 (B6) alleles, causing multiple amino acid changes that have been proposed to be responsible for the differing glycosylation status. To address whether this coding polymorphism recapitulates the disease suppression mediated by the B6 Idd3 allele, we generated knockin mice in which exon 1 of the B6 IL-2 allele replaces the homologous region in the NOD allele. We generated these mice by targeting the NOD allele of NOD/129 F(1) ES cells. IL-2 protein from the knockin mice showed the glycosylation pattern of the B6 IL-2 isoform, confirming that the amino acid differences encoded within exon 1 affect the glycosylation of the IL-2 protein. However, unlike NOD.B6 Idd3 congenic mice, the knockin mice were not protected from T1D. Furthermore, the difference in amino acid sequence in the IL-2 protein did not affect the level of expression of IL-2. This approach provides a general method for the determination of a functional role of a given genomic sequence in a disease process. Further, our result demonstrates that the variants in exon 1 of the IL-2 gene are not responsible for T1D suppression in NOD.B6 Idd3 mice, thereby supporting the hypothesis that variants in the regulatory region affecting expression levels are causative. PMID- 19549861 TI - Translocation experiments with butterflies reveal limits to enhancement of poleward populations under climate change. AB - There is a pressing need to predict how species will change their geographic ranges under climate change. Projections typically assume that temperature is a primary fitness determinant and that populations near the poleward (and upward) range boundary are preadapted to warming. Thus, poleward, peripheral populations will increase with warming, and these increases facilitate poleward range expansions. We tested the assumption that poleward, peripheral populations are enhanced by warming using 2 butterflies (Erynnis propertius and Papilio zelicaon) that co-occur and have contrasting degrees of host specialization and interpopulation genetic differentiation. We performed a reciprocal translocation experiment between central and poleward, peripheral populations in the field and simulated a translocation experiment that included alternate host plants. We found that the performance of both central and peripheral populations of E. propertius were enhanced during the summer months by temperatures characteristic of the range center but that local adaptation of peripheral populations to winter conditions near the range edge could counteract that enhancement. Further, poleward range expansion in this species is prevented by a lack of host plants. In P. zelicaon, the fitness of central and peripheral populations decreased under extreme summer temperatures that occurred in the field at the range center. Performance in this species also was affected by an interaction of temperature and host plant such that host species strongly mediated the fitness of peripheral individuals under differing simulated temperatures. Altogether we have evidence that facilitation of poleward range shifts through enhancement of peripheral populations is unlikely in either study species. PMID- 19549860 TI - Regulation of myocardial ketone body metabolism by the gut microbiota during nutrient deprivation. AB - Studies in mice indicate that the gut microbiota promotes energy harvest and storage from components of the diet when these components are plentiful. Here we examine how the microbiota shapes host metabolic and physiologic adaptations to periods of nutrient deprivation. Germ-free (GF) mice and mice who had received a gut microbiota transplant from conventionally raised donors were compared in the fed and fasted states by using functional genomic, biochemical, and physiologic assays. A 24-h fast produces a marked change in gut microbial ecology. Short chain fatty acids generated from microbial fermentation of available glycans are maintained at higher levels compared with GF controls. During fasting, a microbiota-dependent, Ppar alpha-regulated increase in hepatic ketogenesis occurs, and myocardial metabolism is directed to ketone body utilization. Analyses of heart rate, hydraulic work, and output, mitochondrial morphology, number, and respiration, plus ketone body, fatty acid, and glucose oxidation in isolated perfused working hearts from GF and colonized animals (combined with in vivo assessments of myocardial physiology) revealed that the fasted GF heart is able to sustain its performance by increasing glucose utilization, but heart weight, measured echocardiographically or as wet mass and normalized to tibial length or lean body weight, is significantly reduced in both fasted and fed mice. This myocardial-mass phenotype is completely reversed in GF mice by consumption of a ketogenic diet. Together, these results illustrate benefits provided by the gut microbiota during periods of nutrient deprivation, and emphasize the importance of further exploring the relationship between gut microbes and cardiovascular health. PMID- 19549862 TI - Virtual endocast of Ignacius graybullianus (Paromomyidae, Primates) and brain evolution in early primates. AB - Extant primates are distinctive among mammals in having relatively large brains. As stem primates, Paleogene plesiadapiforms provide direct information relevant to the earliest stages in the evolution of this characteristic. Here we describe a virtual endocast reconstructed from ultra high resolution X-ray computed tomography data for the paromomyid plesiadapiform Ignacius graybullianus (USNM 421608) from the early Eocene of Wyoming. This represents the most complete endocast known for a stem primate, allowing for an unprecedented study of both size and fine details of anatomy. Relative to fossil and extant euprimates, I. graybullianus had large olfactory lobes, but less caudal development of the cerebrum and a poorly demarcated temporal lobe, suggesting more emphasis on olfaction and a less well developed visual system. Although its brain was small compared to those of extant primates, the encephalization quotient of I. graybullianus is higher than that calculated for Paleocene Plesiadapis cookei and overlaps the lower portion of the range documented for fossil euprimates. Comparison to the basal gliroid Rhombomylus suggests that early primates exhibited some expansion of the cerebrum compared to their ancestors. The relatively small brain size of I. graybullianus, an arboreal frugivore, implies that neither arboreality nor frugivory was primarily responsible for the expanded brains of modern primates. However, the contrasts in features related to the visual system between I. graybullianus and fossil and extant euprimates suggest that improvements to these portions of the brain contributed to increases in brain size within Euprimates. PMID- 19549863 TI - Structure and assembly of immature HIV. AB - The major structural components of HIV are synthesized as a 55-kDa polyprotein, Gag. Particle formation is driven by the self-assembly of Gag into a curved hexameric lattice, the structure of which is poorly understood. We used cryoelectron tomography and contrast-transfer-function corrected subtomogram averaging to study the structure of the assembled immature Gag lattice to approximately 17-A resolution. Gag is arranged in the immature virus as a single, continuous, but incomplete hexameric lattice whose curvature is mediated without a requirement for pentameric defects. The resolution of the structure allows positioning of individual protein domains. High-resolution crystal structures were fitted into the reconstruction to locate protein-protein interfaces involved in Gag assembly, and to identify the structural transformations associated with virus maturation. The results of this study suggest a concept for the formation of nonsymmetrical enveloped viruses of variable sizes. PMID- 19549865 TI - Global potential for wind-generated electricity. AB - The potential of wind power as a global source of electricity is assessed by using winds derived through assimilation of data from a variety of meteorological sources. The analysis indicates that a network of land-based 2.5-megawatt (MW) turbines restricted to nonforested, ice-free, nonurban areas operating at as little as 20% of their rated capacity could supply >40 times current worldwide consumption of electricity, >5 times total global use of energy in all forms. Resources in the contiguous United States, specifically in the central plain states, could accommodate as much as 16 times total current demand for electricity in the United States. Estimates are given also for quantities of electricity that could be obtained by using a network of 3.6-MW turbines deployed in ocean waters with depths <200 m within 50 nautical miles (92.6 km) of closest coastlines. PMID- 19549864 TI - DNA wrapping is required for DNA damage recognition in the Escherichia coli DNA nucleotide excision repair pathway. AB - Localized DNA melting may provide a general strategy for recognition of the wide array of chemically and structurally diverse DNA lesions repaired by the nucleotide excision repair (NER) pathway. However, it is not clear what causes such DNA melting and how it is driven. Here, we show a DNA wrapping-melting model supported by results from dynamic monitoring of the key DNA-protein and protein protein interactions involved in the early stages of the Escherichia coli NER process. Using an analytical technique involving capillary electrophoresis coupled with laser-induced fluorescence polarization, which combines a mobility shift assay with conformational analysis, we demonstrate that DNA wrapping around UvrB, mediated by UvrA, is an early event in the damage-recognition process during E. coli NER. DNA wrapping of UvrB was confirmed by Forster resonance energy transfer and fluorescence lifetime measurements. This wrapping did not occur with readily denaturable damaged DNA substrates ("bubble" DNA), suggesting that DNA wrapping of UvrB plays an important role in the induction of DNA melting around the damage site. Analysis of DNA wrapping of mutant UvrB Y96A further suggests that a cooperative interaction between DNA wrapping of UvrA(2)B and contact of the beta-hairpin of UvrB with the bulky damage moiety may be involved in the local DNA melting at the damage site. PMID- 19549866 TI - Protein-free small nuclear RNAs catalyze a two-step splicing reaction. AB - Pre-mRNA splicing is a crucial step in eukaryotic gene expression and is carried out by a highly complex ribonucleoprotein assembly, the spliceosome. Many fundamental aspects of spliceosomal function, including the identity of catalytic domains, remain unknown. We show that a base-paired complex of U6 and U2 small nuclear RNAs, in the absence of the approximately 200 other spliceosomal components, performs a two-step reaction with two short RNA oligonucleotides as substrates that results in the formation of a linear RNA product containing portions of both oligonucleotides. This reaction, which is chemically identical to splicing, is dependent on and occurs in proximity of sequences known to be critical for splicing in vivo. These results prove that the complex formed by U6 and U2 RNAs is a ribozyme and can potentially carry out RNA-based catalysis in the spliceosome. PMID- 19549868 TI - The large contribution of projected HFC emissions to future climate forcing. AB - The consumption and emissions of hydrofluorocarbons (HFCs) are projected to increase substantially in the coming decades in response to regulation of ozone depleting gases under the Montreal Protocol. The projected increases result primarily from sustained growth in demand for refrigeration, air-conditioning (AC) and insulating foam products in developing countries assuming no new regulation of HFC consumption or emissions. New HFC scenarios are presented based on current hydrochlorofluorocarbon (HCFC) consumption in leading applications, patterns of replacements of HCFCs by HFCs in developed countries, and gross domestic product (GDP) growth. Global HFC emissions significantly exceed previous estimates after 2025 with developing country emissions as much as 800% greater than in developed countries in 2050. Global HFC emissions in 2050 are equivalent to 9-19% (CO(2)-eq. basis) of projected global CO(2) emissions in business-as usual scenarios and contribute a radiative forcing equivalent to that from 6-13 years of CO(2) emissions near 2050. This percentage increases to 28-45% compared with projected CO(2) emissions in a 450-ppm CO(2) stabilization scenario. In a hypothetical scenario based on a global cap followed by 4% annual reductions in consumption, HFC radiative forcing is shown to peak and begin to decline before 2050. PMID- 19549867 TI - Extracellular thiol-assisted selenium uptake dependent on the x(c)- cystine transporter explains the cancer-specific cytotoxicity of selenite. AB - The selenium salt selenite (SeO(3)(2-)) is cytotoxic in low to moderate concentrations, with a remarkable specificity for cancer cells resistant to conventional chemotherapy. Our data show that selenium uptake and accumulation, rather than intracellular events, are crucial to the specific selenite cytotoxicity observed in resistant cancer cells. We show that selenium uptake depends on extracellular reduction, and that the extracellular environment is a key factor specific to selenite cytotoxicity. The extracellular reduction is mediated by cysteine, and the efficacy is determined by the uptake of cystine by the x(c)(-) antiporter and secretion of cysteine by multidrug resistance proteins, both of which are frequently overexpressed by resistant cancer cells. This mechanism provides molecular evidence for the existence of an inverse relationship between resistance to conventional chemotherapy and sensitivity to selenite cytotoxicity, and highlights the great therapeutic potential in treating multidrug-resistant cancer. PMID- 19549869 TI - Connexin expression by radial glia-like cells is required for neurogenesis in the adult dentate gyrus. AB - In the adult dentate gyrus, radial glia-like cells represent putative stem cells generating neurons and glial cells. Here, we combined patch-clamp recordings, biocytin filling, immunohistochemistry, single-cell transcript analysis, and mouse transgenics to test for connexin expression and gap junctional coupling of radial glia-like cells and its impact on neurogenesis. Radial glia-like cells were identified in mice expressing EGFP under control of the nestin and gfap promoters. We show that a majority of Radial glia-like cells are coupled and express Cx43. Neuronal precursors were not coupled. Mice lacking Cx30 and Cx43 in GFAP-positive cells displayed almost complete inhibition of proliferation and a significant decline in numbers of radial glia-like cells and granule neurons. Inducible virus-mediated ablation of connexins in the adult hippocampus also reduced neurogenesis. These findings strongly suggest the requirement of connexin expression by radial glia-like cells for intact neurogenesis in the adult brain and point to possible communication pathways of these cells. PMID- 19549870 TI - Engineering crystals of dendritic molecules. AB - A detailed single-crystal X-ray study of conformationally flexible sulfonimide based dendritic molecules with systematically varied molecular architectures was undertaken. Thirteen crystal structures reported in this work include 9 structures of the second-generation dendritic sulfonimides decorated with different aryl groups, 2 compounds bearing branches of both second and first generation, and 2 representatives of the first generation. Analysis of the packing patterns of 9 compounds bearing second-generation branches shows that despite their lack of strong directive functional groups there is a repeatedly reproduced intermolecular interaction mode consisting in an anchor-type packing of complementary second-generation branches of neighbouring molecules. The observed interaction tolerates a wide range of substituents in meta- and para positions of the peripheral arylsulfonyl rings. Quantum chemical calculations of the molecule-molecule interaction energies agree at the qualitative level with the packing preferences found in the crystalline state. The calculations can therefore be used as a tool to rationalize and predict molecular structures with commensurate and non-commensurate branches for programming of different packing modes in crystal. PMID- 19549871 TI - The building blocks of economic complexity. AB - For Adam Smith, wealth was related to the division of labor. As people and firms specialize in different activities, economic efficiency increases, suggesting that development is associated with an increase in the number of individual activities and with the complexity that emerges from the interactions between them. Here we develop a view of economic growth and development that gives a central role to the complexity of a country's economy by interpreting trade data as a bipartite network in which countries are connected to the products they export, and show that it is possible to quantify the complexity of a country's economy by characterizing the structure of this network. Furthermore, we show that the measures of complexity we derive are correlated with a country's level of income, and that deviations from this relationship are predictive of future growth. This suggests that countries tend to converge to the level of income dictated by the complexity of their productive structures, indicating that development efforts should focus on generating the conditions that would allow complexity to emerge to generate sustained growth and prosperity. PMID- 19549872 TI - Optical measurement of mGluR1 conformational changes reveals fast activation, slow deactivation, and sensitization. AB - Metabotropic glutamate receptor (mGluR) activation has been extensively studied under steady-state conditions. However, at central synapses, mGluRs are exposed to brief submillisecond glutamate transients and may not reach steady-state. The lack of information on the kinetics of mGluR activation impairs accurate predictions of their operation during synaptic transmission. Here, we report experiments designed to investigate mGluR kinetics in real-time. We inserted either CFP or YFP into the second intracellular loop of mGluR1beta. When these constructs were coexpressed in PC12 cells, glutamate application induced a conformational change that could be monitored, using fluorescence resonance energy transfer (FRET), with an EC(50) of 7.5 microM. The FRET response was mimicked by the agonist DHPG, abolished by the competitive antagonist MCPG, and partially inhibited by mGluR1-selective allosteric modulators. These results suggest that the FRET response reports active conformations of mGluR1 dimers. The solution exchange at the cell membrane was optimized for voltage-clamped cells by recording the current induced by co-application of 30 mM potassium. When glutamate was applied at increasing concentrations up to 2 mM, the activation time course decreased to a minimum of approximately 10 ms, whereas the deactivation time course remained constant (approximately 50 ms). During long lasting applications, no desensitization was observed. In contrast, we observed a robust sensitization of the FRET response that developed over approximately 400 ms. Activation, deactivation, and sensitization time courses and amplitudes were used to derive a kinetic scheme and rate constants, from which we inferred the EC(50) and frequency dependence of mGluR1 activation under non-steady-state conditions, as occurs during synaptic transmission. PMID- 19549873 TI - Paleocene emergence of elephant relatives and the rapid radiation of African ungulates. AB - Elephants are the only living representatives of the Proboscidea, a formerly diverse mammalian order whose history began with the 55-million years (mys) old Phosphatherium. Reported here is the discovery from the early late Paleocene of Morocco, ca. 60 mys, of the oldest and most primitive elephant relative, Eritherium azzouzorum n.g., n.sp., which is one of the earliest known representatives of modern placental orders. This well supported stem proboscidean is extraordinarily primitive and condylarth-like. It provides the first dental evidence of a resemblance between the proboscideans and African ungulates (paenungulates) on the one hand and the louisinines and early macroscelideans on the other. Eritherium illustrates the origin of the elephant order at a previously unknown primitive stage among paenungulates and "ungulates." The primitive morphology of Eritherium suggests a recent and rapid paenungulate radiation after the Cretaceous-Tertiary boundary, probably favoured by early endemic African paleoecosystems. At a broader scale, Eritherium provides a new old calibration point of the placental tree and supports an explosive placental radiation. The Ouled Abdoun basin, which yields the oldest known African placentals, is a key locality for elucidating phylogeny and early evolution of paenungulates and other related endemic African lineages. PMID- 19549874 TI - Improved spatial learning performance of fat-1 mice is associated with enhanced neurogenesis and neuritogenesis by docosahexaenoic acid. AB - Docosahexaenoic acid (DHA), an n-3 long chain polyunsaturated fatty acid (LC PUFA), highly enriched in the central nervous system, is critical for brain development and function. It has been shown that DHA deficiency impairs cognitive performance whereas DHA supplementation improves the condition. However, the mechanisms underlying the role of DHA in brain development and function remain to be elucidated. By using transgenic fat-1 mice rich in endogenous n-3 PUFA, we show that increased brain DHA significantly enhances hippocampal neurogenesis shown by an increased number of proliferating neurons and neuritogenesis, evidenced by increased density of dendritic spines of CA1 pyramidal neurons in the hippocampus. Concurrently, fat-1 mice exhibit a better spatial learning performance in the Morris water maze compared with control WT littermates. In vitro experiments further demonstrate that DHA promotes differentiation and neurite outgrowth of neuronal cells derived from mouse ES cells and increases the proliferation of cells undergoing differentiation into neuronal lineages from the ES cells. These results together provide direct evidence for a promoting effect of DHA on neurogenesis and neuritogenesis and suggest that this effect may be a mechanism underlying its beneficial effect on behavioral performance. PMID- 19549875 TI - Modeling the immune rheostat of macrophages in the lung in response to infection. AB - In the lung, alternatively activated macrophages (AAM) form the first line of defense against microbial infection. Due to the highly regulated nature of AAM, the lung can be considered as an immunosuppressive organ for respiratory pathogens. However, as infection progresses in the lung, another population of macrophages, known as classically activated macrophages (CAM) enters; these cells are typically activated by IFN-gamma. CAM are far more effective than AAM in clearing the microbial load, producing proinflammatory cytokines and antimicrobial defense mechanisms necessary to mount an adequate immune response. Here, we are concerned with determining the first time when the population of CAM becomes more dominant than the population of AAM. This proposed "switching time" is explored in the context of Mycobacterium tuberculosis (MTb) infection. We have developed a mathematical model that describes the interactions among cells, bacteria, and cytokines involved in the activation of both AAM and CAM. The model, based on a system of differential equations, represents a useful tool to analyze strategies for reducing the switching time, and to generate hypotheses for experimental testing. PMID- 19549876 TI - National differences in gender-science stereotypes predict national sex differences in science and math achievement. AB - About 70% of more than half a million Implicit Association Tests completed by citizens of 34 countries revealed expected implicit stereotypes associating science with males more than with females. We discovered that nation-level implicit stereotypes predicted nation-level sex differences in 8th-grade science and mathematics achievement. Self-reported stereotypes did not provide additional predictive validity of the achievement gap. We suggest that implicit stereotypes and sex differences in science participation and performance are mutually reinforcing, contributing to the persistent gender gap in science engagement. PMID- 19549877 TI - Evidence for food storage and predomestication granaries 11,000 years ago in the Jordan Valley. AB - Food storage is a vital component in the economic and social package that comprises the Neolithic, contributing to plant domestication, increasingly sedentary lifestyles, and new social organizations. Recent excavations at Dhra' near the Dead Sea in Jordan provide strong evidence for sophisticated, purpose built granaries in a predomestication context approximately 11,300-11,175 cal B.P., which support recent arguments for the deliberate cultivation of wild cereals at this time. Designed with suspended floors for air circulation and protection from rodents, they are located between residential structures that contain plant-processing instillations. The granaries represent a critical evolutionary shift in the relationship between people and plant foods, which precedes the emergence of domestication and large-scale sedentary communities by at least 1,000 years. PMID- 19549878 TI - Resistance to age-dependent thymic atrophy in long-lived mice that are deficient in pregnancy-associated plasma protein A. AB - Pregnancy-associated plasma protein A (PAPPA) is a metalloproteinase that controls the tissue availability of insulin-like growth factor (IGF). Homozygous deletion of PAPPA in mice leads to lifespan extension. Since immune function is an important determinant of individual fitness, we examined the natural immune ecology of PAPPA(-/-) mice and their wild-type littermates reared under specific pathogen-free condition with aging. Whereas wild-type mice exhibit classic age dependent thymic atrophy, 18-month-old PAPPA(-/-) mice maintain discrete thymic cortex and medulla densely populated by CD4(+)CD8(+) thymocytes that are capable of differentiating into single-positive CD4 and CD8 T cells. Old PAPPA(-/-) mice have high levels of T cell receptor excision circles, and have bone marrows enriched for subsets of thymus-seeding progenitors. PAPPA(-/-) mice have an overall larger pool of naive T cells, and also exhibit an age-dependent accumulation of CD44(+)CD43(+) memory T cells similar to wild-type mice. However, CD43(+) T cell subsets of old PAPPA(-/-) mice have significantly lower prevalence of 1B11 and S7, glycosylation isoforms known to inhibit T cell activation with normal aging. In bioassays of cell activation, splenic T cells of old PAPPA(-/-) mice have high levels of activation antigens and cytokine production, and also elicit Ig production by autologous B cells at levels equivalent to young wild type mice. These data suggest an IGF-immune axis of healthy longevity. Controlling the availability of IGF in the thymus by targeted manipulation of PAPPA could be a way to maintain immune homeostasis during postnatal development and aging. PMID- 19549880 TI - TARP modulation of synaptic AMPA receptor trafficking and gating depends on multiple intracellular domains. AB - Previous work has established stargazin and its related family of transmembrane AMPA receptor regulatory proteins (TARPs) as auxiliary subunits of AMPA receptors (AMPARs) that control synaptic strength both by targeting AMPARs to synapses through an intracellular PDZ-binding motif and by modulating their gating through an extracellular domain. However, TARPs gamma-2 and gamma-8 differentially regulate the synaptic targeting of AMPARs, despite having identical PDZ-binding motifs. Here, we investigate the structural elements that contribute to this functional difference between TARP subtypes by using domain transplantation and truncation. We identify a component of synaptic AMPAR trafficking that is independent of the TARP C-terminal PDZ-binding motif, and we establish previously uncharacterized roles for the TARP intracellular N terminus, loop, and C terminus in modulating both the trafficking and gating of synaptic AMPARs. PMID- 19549879 TI - An engineered substance P variant for receptor-mediated delivery of synthetic antibodies into tumor cells. AB - We have developed and tested a robust delivery method for the transport of proteins to the cytoplasm of mammalian cells without compromising the integrity of the cell membrane. This receptor-mediated delivery (RMD) technology utilizes a variant of substance P (SP), a neuropeptide that is rapidly internalized upon interaction with the neurokinin-1 receptor (NK1R). Cargos in the form of synthetic antibody fragments (sABs) were conjugated to the engineered SP variant (SPv) and efficiently internalized by NK1R-expressing cells. The sABs used here were generated to bind specific conformational forms of actin. The internalized proteins appear to escape the endosome and retain their binding activity within the cells as demonstrated by co-localization with the actin cytoskeleton. Further, since the NK1R is over-expressed in many cancers, SPv-mediated delivery provides a highly specific method for therapeutic utilization of affinity reagents targeting intracellular processes in diseased tissue. PMID- 19549881 TI - The Mo-Se active site of nicotinate dehydrogenase. AB - Nicotinate dehydrogenase (NDH) from Eubacterium barkeri is a molybdoenzyme catalyzing the hydroxylation of nicotinate to 6-hydroxynicotinate. Reactivity of NDH critically depends on the presence of labile (nonselenocysteine) selenium with an as-yet-unidentified form and function. We have determined the crystal structure of NDH and analyzed its active site by multiple wavelengths anomalous dispersion methods. We show that selenium is bound as a terminal Mo=Se ligand to molybdenum and that it occupies the position of the terminal sulfido ligand in other molybdenum hydroxylases. The role of selenium in catalysis has been assessed by model calculations, which indicate an acceleration of the critical hydride transfer from the substrate to the selenido ligand in the course of substrate hydroxylation when compared with an active site containing a sulfido ligand. The MoO(OH)Se active site of NDH shows a novel type of utilization and reactivity of selenium in nature. PMID- 19549882 TI - Neuromedin S and U. PMID- 19549883 TI - Finally! A model for progesterone receptor action in normal human breast. PMID- 19549884 TI - Sexual differentiation of the nervous system: where the action is. PMID- 19549885 TI - Uncoupling modifier genes from uncoupling protein 2 in pancreatic beta-cells. PMID- 19549886 TI - MDA-MB-435 and M14 cell lines: identical but not M14 melanoma? AB - A controversy has arisen over the past several years about the true origin of the human MDA-MB-435 cell line. Originally described as a human breast cancer cell line, subsequent expression array studies instead suggested a gene expression profile consistent with a melanoma origin. Subsequent karyotype and comparative genomic hybridization studies supported the idea that current stocks of both MDA MB-435 cells and M14 melanoma cells must be identical cell lines, and the conclusion was drawn that both cell lines were in fact M14 melanoma cells. However, an alternate conclusion based on these data is that both cell lines are indeed identical, but are in fact MDA-MB-435 breast cancer cells. There is evidence that many cell lines can display "lineage infidelity" and that assignment to tissue type is unreliably made based on expression patterns. Evidence from the literature is presented here that is inconsistent with both lines being of M14 melanoma origin, but rather is consistent with both cell lines being of MDA-MB-435 breast cancer origin. PMID- 19549887 TI - Aneuploidy: cancer's fatal flaw? AB - Aneuploidy is a characteristic of cancer, with greater than 90% of all solid tumors in humans carrying an aberrant karyotype. Yet, whether or how this condition contributes to tumorigenesis is not understood. Here we summarize our recent findings on the effects of aneuploidy on cell physiology and proliferation. These studies suggest that aneuploidy puts significant stress on the cell, which responds to this condition in what can be viewed as an aneuploidy stress response. We will discuss how our results may bear on our understanding of the role of this condition in tumorigenesis and how they may provide new opportunities for treatment of the disease. PMID- 19549888 TI - Functional phosphodiesterase 11A mutations may modify the risk of familial and bilateral testicular germ cell tumors. AB - Inactivating germline mutations in phosphodiesterase 11A (PDE11A) have been implicated in adrenal tumor susceptibility. PDE11A is highly expressed in endocrine steroidogenic tissues, especially the testis, and mice with inactivated Pde11a exhibit male infertility, a known testicular germ cell tumor (TGCT) risk factor. We sequenced the PDE11A gene-coding region in 95 patients with TGCT from 64 unrelated kindreds. We identified 8 nonsynonymous substitutions in 20 patients from 15 families: four (R52T, F258Y, G291R, and V820M) were newly recognized, three (R804H, R867G, and M878V) were functional variants previously implicated in adrenal tumor predisposition, and one (Y727C) was a known polymorphism. We compared the frequency of these variants in our patients to unrelated controls that had been screened and found negative for any endocrine diseases: only the two previously reported variants, R804H and R867G, known to be frequent in general population, were detected in these controls. The frequency of all PDE11A gene variants (combined) was significantly higher among patients with TGCT (P = 0.0002), present in 19% of the families of our cohort. Most variants were detected in the general population, but functional studies showed that all these mutations reduced PDE activity, and that PDE11A protein expression was decreased (or absent) in TGCT samples from carriers. This is the first demonstration of the involvement of a PDE gene in TGCT, although the cyclic AMP signaling pathway has been investigated extensively in reproductive organ function and their diseases. In conclusion, we report that PDE11A-inactivating sequence variants may modify the risk of familial and bilateral TGCT. PMID- 19549890 TI - Building scientific progress without borders: nanobiology and nanomedicine in China and the U.S. PMID- 19549889 TI - A "vascular normalization index" as potential mechanistic biomarker to predict survival after a single dose of cediranib in recurrent glioblastoma patients. AB - Early imaging or blood biomarkers of tumor response are desperately needed to customize antiangiogenic therapy for cancer patients. Anti-vascular endothelial growth factor (VEGF) therapy can "normalize" brain tumor vasculature by decreasing vessel diameter and permeability, and thinning the abnormally thick basement membrane. We hypothesized that the extent of vascular normalization will be predictive of outcome of anti-VEGF therapy in glioblastoma. We used advanced magnetic resonance imaging methods to monitor vascular parameters and treatment response in 31 recurrent glioblastoma patients enrolled in a phase II trial of cediranib, an oral pan-VEGF receptor tyrosine kinase inhibitor. We evaluated the correlation between clinical outcome and magnetic resonance imaging-measured changes in vascular permeability/flow (i.e., K(trans)) and in microvessel volume, and the change of circulating collagen IV levels, all after a single dose of cediranib. Here, we show that evaluation of biomarkers as early as after one day of anti-VEGF therapy with cediranib is predictive of response in patients with recurrent glioblastoma. Changes in K(trans), microvessel volume, and circulating collagen IV correlated with duration of overall survival and/or progression-free survival (P < 0.05). When we combined these three parameters into a "vascular normalization index," we found that it closely associated with overall survival (rho = 0.54; P = 0.004) and progression-free survival (rho = 0.6; P = 0.001). The vascular normalization index described here should be validated in randomized clinical trials. PMID- 19549891 TI - The inhibitor of apoptosis protein Livin (ML-IAP) plays a dual role in tumorigenicity. AB - The inhibitor of apoptosis protein (IAP) family can inhibit apoptosis induced by a variety of stimuli. We and others previously described the IAP Livin (ML-IAP). We found that Livin is unique among the IAP members as, on a strong apoptotic stimulus, it is specifically cleaved by caspases to produce a truncated protein with paradoxical proapoptotic activity (tLivin). We also showed that Livin encodes two splicing variants, termed Livin alpha and beta, with diverse antiapoptotic effects in vitro. In this study, we compared the Livin isoforms in vivo. An animal model was established and the effects of Livin alpha and beta on the initiation and development of tumors were compared. In the animal model, Livin alpha promotes tumor initiation in comparison with control. Interestingly, the growth of tumors originating from cells expressing Livin beta was inhibited. In these tumors, Livin beta was cleaved and produced a high level of the proapoptotic tLivin beta that repressed tumor development. When we eliminated the proapoptotic effect of Livin beta by point mutations, the resulting antiapoptotic Livin beta mutants contributed to tumor progression. In terms of mechanism, we show that Livin beta tumors develop only in mice lacking natural killer (NK) cell activity. Thus, from the animal model, we can conclude that Livin plays a major role in tumorigenicity and that NK cells induce cleavage of Livin to its proapoptotic truncated protein that in turn inhibits tumor growth. Therefore, Livin and tLivin may serve as potential targets for cancer therapy. PMID- 19549892 TI - Prostaglandin F2alpha-F-prostanoid receptor signaling promotes neutrophil chemotaxis via chemokine (C-X-C motif) ligand 1 in endometrial adenocarcinoma. AB - The prostaglandin F(2alpha) (PGF(2alpha)) receptor (FP) is elevated in endometrial adenocarcinoma. This study found that PGF(2alpha) signaling via FP regulates expression of chemokine (C-X-C motif) ligand 1 (CXCL1) in endometrial adenocarcinoma cells. Expression of CXCL1 and its receptor, CXCR2, are elevated in cancer tissue compared with normal endometrium and localized to glandular epithelium, endothelium, and stroma. Treatment of Ishikawa cells stably transfected with the FP receptor (FPS cells) with 100 nmol/L PGF(2alpha) increased CXCL1 promoter activity, mRNA, and protein expression, and these effects were abolished by cotreatment of cells with FP antagonist or chemical inhibitors of Gq, epidermal growth factor receptor, and extracellular signal regulated kinase. Similarly, CXCL1 was elevated in response to 100 nmol/L PGF(2alpha) in endometrial adenocarcinoma explant tissue. CXCL1 is a potent neutrophil chemoattractant. The expression of CXCR2 colocalized to neutrophils in endometrial adenocarcinoma and increased neutrophils were present in endometrial adenocarcinoma compared with normal endometrium. Conditioned media from PGF(2alpha)-treated FPS cells stimulated neutrophil chemotaxis, which could be abolished by CXCL1 protein immunoneutralization of the conditioned media or antagonism of CXCR2. Finally, xenograft tumors in nude mice arising from inoculation with FPS cells showed increased neutrophil infiltration compared with tumors arising from wild-type cells or following treatment of mice bearing FPS tumors with CXCL1-neutralizing antibody. In conclusion, our results show a novel PGF(2alpha)-FP pathway that may regulate the inflammatory microenvironment in endometrial adenocarcinoma via neutrophil chemotaxis. PMID- 19549893 TI - Evaluation of the 8q24 prostate cancer risk locus and MYC expression. AB - Polymorphisms at 8q24 are robustly associated with prostate cancer risk. The risk variants are located in nonprotein coding regions and their mechanism has not been fully elucidated. To further dissect the function of this locus, we tested two hypotheses: (a) unannotated microRNAs (miRNA) are transcribed in the region, and (b) this region is a cis-acting enhancer. Using next generation sequencing, 8q24 risk regions were interrogated for known and novel miRNAs in histologically normal radical prostatectomy tissue. We also evaluated the association between the risk variants and transcript levels of multiple genes, focusing on the proto oncogene, MYC. RNA expression was measured in histologically normal and tumor tissue from 280 prostatectomy specimens (from 234 European American and 46 African American patients), and paired germline DNA from each individual was genotyped for six 8q24 risk single nucleotide polymorphisms. No evidence was found for significant miRNA transcription within 8q24 prostate cancer risk loci. Likewise, no convincing association between RNA expression and risk allele status was detected in either histologically normal or tumor tissue. To our knowledge, this is one of the first and largest studies to directly assess miRNA in this region and to systematically measure MYC expression levels in prostate tissue in relation to inherited risk variants. These data will help to direct the future study of this risk locus. PMID- 19549894 TI - Beta1 integrin cytoplasmic variants differentially regulate expression of the antiangiogenic extracellular matrix protein thrombospondin 1. AB - Beta(1) integrins play an important role in regulating cell proliferation and survival. Using small interfering RNA or an inhibitory antibody to beta(1), we show here that, in vivo, beta(1) integrins are essential for prostate cancer growth. Among the five known beta(1) integrin cytoplasmic variants, two have been shown to differentially affect prostate cell functions. The beta(1A) variant promotes normal and cancer cell proliferation, whereas the beta(1C) variant, which is down-regulated in prostate cancer, inhibits tumor growth and appears to have a dominant effect on beta(1A). To investigate the mechanism by which beta(1C) inhibits the tumorigenic potential of beta(1A), we analyzed changes in gene expression in cells transfected with either beta(1C) or beta(1A). The results show that beta(1C) expression increases the levels of an extracellular matrix protein, thrombospondin 1 (TSP1), an angiogenesis inhibitor. TSP1 protein levels are increased upon beta(1C) expression in prostate cancer cells as well as in beta(1)-null GD25 cells. We show that TSP1 does not affect proliferation, apoptosis, or anchorage-independent growth of prostate cancer cells. In contrast, the newly synthesized TSP1, secreted by prostate cancer cells expressing beta(1C), prevents proliferation of endothelial cells. In conclusion, our novel findings indicate that expression of the beta(1C) integrin variant in prostate glands prevents cancer progression by up-regulation of TSP1 levels and inhibition of angiogenesis. PMID- 19549895 TI - Cell survival under stress is enhanced by a mitochondrial ATP-binding cassette transporter that regulates hemoproteins. AB - The ATP-binding cassette (ABC) transporter ABCB6 localizes to the mitochondria, where it imports porphyrins and up-regulates de novo porphyrin synthesis. If ABCB6 also increases the intracellular heme concentration, it may broadly affect the regulation and physiology of cellular hemoproteins. We tested whether the ability of ABCB6 to accelerate de novo porphyrin biosynthesis alters mitochondrial and extramitochondrial heme levels. ABCB6 overexpression increased the quantity of cytosolic heme but did not affect mitochondrial heme levels. We then tested whether the increased extramitochondrial heme would increase the concentration and/or activity of cellular hemoproteins (hemoglobin, catalase, and cytochrome c oxidase). ABCB6 overexpression increased the activity and quantity of hemoproteins found in several subcellular compartments, and reduction of ABCB6 function (by small interfering RNA or knockout) reversed these findings. In complementary studies, suppression of ABCB6 expression sensitized cells to stress induced by peroxide and cyanide, whereas overexpression of ABCB6 protected against both stressors. Our findings show that the ability of ABCB6 to increase cytosolic heme levels produces phenotypic changes in hemoproteins that protect cells from certain stresses. Collectively, these findings have implications for the health and survival of both normal and abnormal cells, which rely on heme for multiple cellular processes. PMID- 19549897 TI - Xenoestrogen-induced epigenetic repression of microRNA-9-3 in breast epithelial cells. AB - Early exposure to xenoestrogens may predispose to breast cancer risk later in adult life. It is likely that long-lived, self-regenerating epithelial progenitor cells are more susceptible to these exposure injuries over time and transmit the injured memory through epigenetic mechanisms to their differentiated progeny. Here, we used progenitor-containing mammospheres as an in vitro exposure model to study this epigenetic effect. Expression profiling identified that, relative to control cells, 9.1% of microRNAs (82 of 898 loci) were altered in epithelial progeny derived from mammospheres exposed to a synthetic estrogen, diethylstilbestrol. Repressive chromatin marks, trimethyl Lys27 of histone H3 (H3K27me3) and dimethyl Lys9 of histone H3 (H3K9me2), were found at a down regulated locus, miR-9-3, in epithelial cells preexposed to diethylstilbestrol. This was accompanied by recruitment of DNA methyltransferase 1 that caused an aberrant increase in DNA methylation of its promoter CpG island in mammosphere derived epithelial cells on diethylstilbestrol preexposure. Functional analyses suggest that miR-9-3 plays a role in the p53-related apoptotic pathway. Epigenetic silencing of this gene, therefore, reduces this cellular function and promotes the proliferation of breast cancer cells. Promoter hypermethylation of this microRNA may be a hallmark for early breast cancer development, and restoration of its expression by epigenetic and microRNA-based therapies is another viable option for future treatment of this disease. PMID- 19549898 TI - MUC1, a new hypoxia inducible factor target gene, is an actor in clear renal cell carcinoma tumor progression. AB - The hypoxia inducible factor (HIF) signaling pathway is known as the main renal carcinogenetic pathway. MUC1, an O-glycoprotein membrane-bound mucin, is overexpressed in clear renal cell carcinomas (cRCC) with correlation to two major prognostic factors: tumor-node-metastasis stage and nuclear Furhman grade. We questioned whether there is a direct link between the HIF pathway and MUC1 overexpression in renal tumors. Interestingly, we observed concomitant increase of HIF-1alpha and MUC1 in metastatic cRCC group versus nonmetastatic cRCC group. Using different renal cell models and small interfering RNA assays targeting either HIF-1alpha or YC-1, a HIF-1 pharmacologic inhibitor, we showed induction of MUC1 expression under hypoxia by a HIF-dependent mechanism. Chromatin immunoprecipitation assay showed a direct binding of HIF-1alpha at the MUC1 promoter. In addition, combined site-directed mutagenesis and gel shift assay allowed the identification of two functional putative hypoxia responsive elements at -1488/-1485 and at -1510/-1507 in the promoter. Using a rat kidney model of ischemia/reperfusion, we confirmed in vivo that clamping renal pedicle for 1 hour followed by 2 hours of reperfusion induced increased MUC1 expression. Furthermore, MUC1 knockdown induced significant reduction of invasive and migration properties of renal cancer cells under hypoxia. Altogether, these results show that MUC1 is directly regulated by HIF-1alpha and affects the invasive and migration properties of renal cancer cells. Thus, MUC1 could serve as a potential therapeutic target in cRCC. PMID- 19549896 TI - Therapeutics by cytotoxic metabolite accumulation: pemetrexed causes ZMP accumulation, AMPK activation, and mammalian target of rapamycin inhibition. AB - Pemetrexed represents the first antifolate cancer drug to be approved by the Food and Drug Administration in 20 years; it is currently in widespread use for first line therapy of mesothelioma and non-small cell lung cancer. Pemetrexed has more than one site of action; the primary site is thymidylate synthase. We now report that the secondary target is the downstream folate-dependent enzyme in de novo purine synthesis, aminoimidazolecarboxamide ribonucleotide formyltransferase (AICART). The substrate of the AICART reaction, ZMP, accumulated in intact pemetrexed-inhibited tumor cells, identifying AICART as the step in purine synthesis that becomes rate-limiting after drug treatment. The accumulating ZMP causes an activation of AMP-activated protein kinase with subsequent inhibition of the mammalian target of rapamycin (mTOR) and hypophosphorylation of the downstream targets of mTOR that control initiation of protein synthesis and cell growth. We suggest that the activity of pemetrexed against human cancers is a reflection of its direct inhibition of folate-dependent target proteins combined with prolonged inhibition of the mTOR pathway secondary to accumulation of ZMP. PMID- 19549899 TI - Plasminogen kringle 5 induces apoptosis of brain microvessel endothelial cells: sensitization by radiation and requirement for GRP78 and LRP1. AB - Recombinant plasminogen kringle 5 (rK5) has been shown to induce apoptosis of dermal microvessel endothelial cells (MvEC) in a manner that requires glucose regulated protein 78 (GRP78). As we are interested in antiangiogenic therapy for glioblastoma tumors, and the effectiveness of antiangiogenic therapy can be enhanced when combined with radiation, we investigated the proapoptotic effects of rK5 combined with radiation on brain MvEC. We found that rK5 treatment of brain MvEC induced apoptosis in a dose- and time-dependent manner and that prior irradiation significantly sensitized (500-fold) the cells to rK5-induced apoptosis. The rK5-induced apoptosis of both unirradiated and irradiated MvEC required expression of GRP78 and the low-density lipoprotein receptor-related protein 1 (LRP1), a scavenger receptor, based on down-regulation studies with small interfering RNA, and blocking studies with either a GRP78 antibody or a competitive inhibitor of ligand binding to LRP1. Furthermore, p38 mitogen activated protein kinase was found to be a necessary downstream effector for rK5 induced apoptosis. These data suggest that irradiation sensitizes brain MvEC to the rK5-induced apoptosis and that this signal requires LRP1 internalization of GRP78 and the activation of p38 mitogen-activated protein kinase. Our findings suggest that prior irradiation would have a dose-sparing effect on rK5 antiangiogenic therapy for brain tumors and further suggest that the effects of rK5 would be tumor specific, as the expression of GRP78 protein is up-regulated on the brain MvEC in glioblastoma tumor biopsies compared with the normal brain. PMID- 19549901 TI - Suppression of nonhomologous end joining repair by overexpression of HMGA2. AB - Understanding the molecular details associated with aberrant high mobility group A2 (HMGA2) gene expression is key to establishing the mechanism(s) underlying its oncogenic potential and effect on the development of therapeutic strategies. Here, we report the involvement of HMGA2 in impairing DNA-dependent protein kinase (DNA-PK) during the nonhomologous end joining (NHEJ) process. We showed that HMGA2-expressing cells displayed deficiency in overall and precise DNA end joining repair and accumulated more endogenous DNA damage. Proper and timely activation of DNA-PK, consisting of Ku70, Ku80, and DNA-PKcs subunits, is essential for the repair of DNA double strand breaks (DSB) generated endogenously or by exposure to genotoxins. In cells overexpressing HMGA2, accumulation of histone 2A variant X phosphorylation at Ser-139 (gamma-H2AX) was associated with hyperphosphorylation of DNA-PKcs at Thr-2609 and Ser-2056 before and after the induction of DSBs. Also, the steady-state complex of Ku and DNA ends was altered by HMGA2. Microirradiation and real-time imaging in living cells revealed that HMGA2 delayed the release of DNA-PKcs from DSB sites, similar to observations found in DNA-PKcs mutants. Moreover, HMGA2 alone was sufficient to induce chromosomal aberrations, a hallmark of deficiency in NHEJ-mediated DNA repair. In summary, a novel role for HMGA2 to interfere with NHEJ processes was uncovered, implicating HMGA2 in the promotion of genome instability and tumorigenesis. PMID- 19549902 TI - Protein kinase Cdelta activates RelA/p65 and nuclear factor-kappaB signaling in response to tumor necrosis factor-alpha. AB - Nuclear factor-kappaB (NF-kappaB) is tightly modulated by IkappaB kinases and IkappaBalpha in the cytoplasm. On stimulation, NF-kappaB translocates into the nucleus to initiate transcription; however, regulation of its transcriptional activity remains obscure. Here, we show that protein kinase C (PKC) delta controls the main subunit of NF-kappaB, RelA/p65. On exposure to tumor necrosis factor-alpha (TNF-alpha), the expression of RelA/p65 target genes such as IkappaBalpha, RelB, and p100/p52 is up-regulated in a PKCdelta-dependent manner. The results also show that PKCdelta is targeted to the nucleus and forms a complex with RelA/p65 following TNF-alpha exposure. Importantly, kinase activity of PKCdelta is required for RelA/p65 transactivation. In concert with these results, PKCdelta activates RelA/p65 for its occupancy to target-gene promoters, including IkappaBalpha and p100/p52. Moreover, functional analyses show that inhibition of PKCdelta is associated with substantial attenuation of NF-kappaB activity in response to TNF-alpha. These findings provide evidence that PKCdelta orchestrates RelA/p65 transactivation, a requisite for NF-kappaB signaling pathway in the nucleus. PMID- 19549900 TI - Microcephalin regulates BRCA2 and Rad51-associated DNA double-strand break repair. AB - Microcephalin (MCPH1) is a BRCA1 COOH terminal (BRCT) domain containing protein involved in the cellular response to DNA damage that has been implicated in autosomal recessive primary microcephaly. MCPH1 is recruited to sites of DNA double-strand breaks by phosphorylated histone H2AX (gammaH2AX), but the mechanism by which MCPH1 contributes to the repair process remains to be determined. Here, we show that MCPH1 binds to BRCA2 and regulates the localization of BRCA2 and Rad51 at sites of DNA damage. The interaction occurs through the NH(2) terminus of BRCA2 and the COOH terminal BRCT domains of MCPH1. Disruption of the interaction between MCPH1 and BRCA2 has no effect on the ability of BRCA2 to form a complex with Rad51 but is associated with substantially reduced levels of both BRCA2 and Rad51 at sites of DNA double strand breaks. Uncoupling of MCPH1 from BRCA2 also interferes with Rad51 dependent and BRCA2-dependent homologous recombination repair activity. These results suggest that the role of MCPH1 in the DNA damage response is in part associated with the ability to localize BRCA2 to sites of DNA double-stand breaks. PMID- 19549904 TI - Lessons from an aggressive cancer: evolutionary dynamics in esophageal carcinoma. AB - Rapid progression to metastatic disease and an intrinsic resistance to any type of systemic therapy are hallmarks of aggressive solid cancers. The molecular basis for this phenotype is not clear. A detailed study of the somatic progression from local to early systemic esophageal cancer revealed rapid diversification of cancer cells isolated from various sites, but also evidence for early clonal expansion. These findings have implications for diagnostic pathology and therapeutic decision making. PMID- 19549903 TI - The liver is a site for tumor-induced myeloid-derived suppressor cell accumulation and immunosuppression. AB - Tumor-induced immunosuppression plays a key role in tumor evasion of the immune system. A key cell population recognized as myeloid-derived suppressor cells (MDSC) contributes and helps orchestrate this immunosuppression. MDSC can interact with T cells, macrophages, and natural killer cells to create an environment favorable for tumor progression. In various tumor models, their presence at high levels has been reported in the bone marrow, blood, spleen, and tumor. We report for the first time that MDSC accumulate and home to the liver in addition to the other organs. Liver MDSC suppress T cells and accumulate to levels comparable with splenic MDSC. Additionally, hematopoiesis in the liver contributes to the dramatic expansion of MDSC in this organ. Furthermore, MDSC in the liver interact with macrophages, also known as Kupffer cells, and cause their up-regulation of PD-L1, a negative T-cell costimulatory molecule. The liver is thus an organ in which MDSC accumulate and can contribute to immunosuppression directly and indirectly. MDSC play a role in various pathologic states in addition to cancer, and these results contribute to our understanding of their biology and interactions with immune-related cells. PMID- 19549905 TI - Differential response of glioma cells to FOXO1-directed therapy. AB - Gliomas are the most common adult primary brain tumors, and the most malignant form, glioblastoma multiforme, is invariably fatal. The phosphatidylinositol 3 kinase (PI3K)-Akt signaling pathway is altered in most glioblastoma multiforme. PTEN, an important negative regulator of the PI3K-Akt pathway, is also commonly mutated in glioma, leading to constitutive activation of Akt. One ultimate consequence is phosphorylation and inactivation of FOXO forkhead transcription factors that regulate genes involved in apoptosis, cell cycle arrest, nutrient availability, DNA repair, stress, and angiogenesis. We tested the ability of a mutant FOXO1 factor that is not subject to Akt phosphorylation to overcome dysregulated PI3K-Akt signaling in two PTEN-null glioma cell lines, U87 and U251. Adenovirus-mediated gene transfer of the mutant FOXO1 successfully restored cell cycle arrest and induced cell death in vitro and prolonged survival in vivo in xenograft models of human glioma (33% survival at 1 year of animals bearing U251 tumors). However, U87 were much more resistant than U251 to mutant FOXO1-induced death, showing evidence of increased nuclear export and Akt-independent phosphorylation of FOXO1 at S249. A cyclin-dependent kinase 2 inhibitor decreased phosphorylation of S249 and rendered U87 cells significantly more susceptible to mutant FOXO1-induced death. Our results indicate that targeting FOXO1, which is at the convergence point of several growth factor receptor tyrosine kinase pathways, can effectively induce glioma cell death and inhibit tumor growth. They also highlight the importance of Akt-independent phosphorylation events in the nuclear export of FOXO1. PMID- 19549907 TI - Targeted in vivo imaging of integrin alphavbeta6 with an improved radiotracer and its relevance in a pancreatic tumor model. AB - The cell surface receptor alpha(v)beta(6) is epithelial specific, and its expression is tightly regulated; it is low or undetectable in adult tissues but has been shown to be increased in many different cancers, including pancreatic, cervical, lung, and colon cancers. Studies have described alpha(v)beta(6) as a prognostic biomarker linked to poor survival. We have recently shown the feasibility of imaging alpha(v)beta(6) in vivo by positron emission tomography (PET) using the peptide [(18)F]FBA-A20FMDV2. Here, we describe improved alpha(v)beta(6) imaging agents and test their efficacy in a mouse model with endogenous alpha(v)beta(6) expression. The modified compounds maintained high affinity for alpha(v)beta(6) and >1,000-fold selectivity over related integrins (by ELISA) and showed significantly improved alpha(v)beta(6)-dependent binding in cell-based assays (>60% binding versus <10% for [(18)F]FBA-A20FMDV2). In vivo studies using either a melanoma cell line (transduced alpha(v)beta(6) expression) or the BxPC-3 human pancreatic carcinoma cell line (endogenous alpha(v)beta(6) expression) revealed that the modified compounds showed significantly improved tumor retention. This, along with good clearance of nonspecifically bound activity, particularly for the new radiotracer [(18)F]FBA-PEG(28)-A20FMDV2, resulted in improved PET imaging. Tumor/pancreas and tumor/blood biodistribution ratios of >23:1 and >47:1, respectively, were achieved at 4 hours. Significantly, [(18)F]FBA-PEG(28)-A20FMDV2 was superior to 2-[(18)F]fluoro-2-deoxy-d-glucose ([(18)F]FDG) in imaging the BxPC-3 tumors. Pancreatic ductal adenocarcinoma is highly metastatic and current preoperative evaluation of resectability using noninvasive imaging has limited success, with most patients having metastases at time of surgery. The fact that these tumors express alpha(v)beta(6) suggests that this probe has significant potential for the in vivo detection of this malignancy, thus having important implications for patient care and therapy. PMID- 19549906 TI - Overexpression of DPAGT1 leads to aberrant N-glycosylation of E-cadherin and cellular discohesion in oral cancer. AB - Cancer cells are frequently characterized by aberrant increases in protein N glycosylation and by disruption of E-cadherin-mediated adherens junctions. The relationship between altered N-glycosylation and loss of E-cadherin adhesion in cancer, however, remains unclear. Previously, we reported that complex N-glycans on the extracellular domains of E-cadherin inhibited the formation of mature adherens junctions. Here, we examined whether dysregulated N-glycosylation was one of the underlying causes for cellular discohesion in oral cancer. We show that dense cultures of human salivary epidermoid carcinoma A253 cells exhibited elevated expression of DPAGT1, the gene that initiates protein N-glycosylation. Overexpression of DPAGT1 correlated with the production of E-cadherin-bearing complex N-glycans in nascent adherens junctions. Partial inhibition of DPAGT1 with small interfering RNA reduced the complex N-glycans of E-cadherin and increased the abundance of alpha-catenin and stabilizing proteins in adherens junctions. This was associated with the assembly of functional tight junctions. The inverse relationship between DPAGT1 expression and intercellular adhesion was a feature of oral squamous cell carcinoma. Oral squamous cell carcinomas displayed overexpression of DPAGT1 that correlated with diminished localization of E-cadherin and alpha-catenin at the sites of adherens junctions. Our studies show for the first time that DPAGT1 is an upstream regulator of E-cadherin N glycosylation status and adherens junction composition and suggest that dysregulation of DPAGT1 causes disturbances in intercellular adhesion in oral cancer. PMID- 19549908 TI - A ruthenium-containing organometallic compound reduces tumor growth through induction of the endoplasmic reticulum stress gene CHOP. AB - Cisplatin-derived anticancer therapy has been used for three decades despite its side effects. Other types of organometallic complexes, namely, some ruthenium derived compounds (RDC), which would display cytotoxicity through different modes of action, might represent alternative therapeutic agents. We have studied both in vitro and in vivo the biological properties of RDC11, one of the most active compounds of a new class of RDCs that contain a covalent bond between the ruthenium atom and a carbon. We showed that RDC11 inhibited the growth of various tumors implanted in mice more efficiently than cisplatin. Importantly, in striking contrast with cisplatin, RDC11 did not cause severe side effects on the liver, kidneys, or the neuronal sensory system. We analyzed the mode of action of RDC11 and showed that RDC11 interacted poorly with DNA and induced only limited DNA damages compared with cisplatin, suggesting alternative transduction pathways. Indeed, we found that target genes of the endoplasmic reticulum stress pathway, such as Bip, XBP1, PDI, and CHOP, were activated in RDC11-treated cells. Induction of the transcription factor CHOP, a crucial mediator of endoplasmic reticulum stress apoptosis, was also confirmed in tumors treated with RDC11. Activation of CHOP led to the expression of several of its target genes, including proapoptotic genes. In addition, the silencing of CHOP by RNA interference significantly reduced the cytotoxicity of RDC11. Altogether, our results led us to conclude that RDC11 acts by an atypical pathway involving CHOP and endoplasmic reticulum stress, and thus might provide an interesting alternative for anticancer therapy. PMID- 19549909 TI - Expression of WSX1 in tumors sensitizes IL-27 signaling-independent natural killer cell surveillance. AB - It is well known that the interleukin (IL)-27 receptor WSX1 is expressed in immune cells and induces an IL-27-dependent immune response. Opposing this conventional dogma, this study reveals a much higher level of WSX1 expression in multiple types of epithelial tumor cells when compared with normal epithelial cells. Expression of exogenous WSX1 in epithelial tumor cells suppresses tumorigenicity in vitro and inhibits tumor growth in vivo. Different from the role of WSX1 in immune cells, the antitumor activity of WSX1 in epithelial tumor cells is independent of IL-27 signaling but is mainly dependent on natural killer (NK) cell surveillance. Deficiency of either the IL-27 subunit EBV-induced gene 3 or the IL-27 receptor WSX1 in the host animals had no effect on tumor growth inhibition induced by WSX1 expression in tumor cells. Expression of WSX1 in epithelial tumor cells enhances NK cell cytolytic activity against tumor cells, whereas the absence of functional NK cells impairs the WSX1-mediated inhibition of epithelial tumor growth. The underlying mechanism by which WSX1 expression in tumor cells enhances NK cytolytic activity is dependent on up-regulation of NKG2D ligand expression. Our results reveal an IL-27-independent function of WSX1: sensitizing NK cell-mediated antitumor surveillance via a NKG2D-dependent mechanism. PMID- 19549910 TI - miR-15a and miR-16 are implicated in cell cycle regulation in a Rb-dependent manner and are frequently deleted or down-regulated in non-small cell lung cancer. AB - MicroRNAs (miRNA) are negative regulators of gene expression at the posttranscriptional level, which are involved in tumorigenesis. Two miRNAs, miR 15a and miR-16, which are located at chromosome 13q14, have been implicated in cell cycle control and apoptosis, but little information is available about their role in solid tumors. To address this question, we established a protocol to quantify miRNAs from laser capture microdissected tissues. Here, we show that miR 15a/miR-16 are frequently deleted or down-regulated in squamous cell carcinomas and adenocarcinomas of the lung. In these tumors, expression of miR-15a/miR-16 inversely correlates with the expression of cyclin D1. In non-small cell lung cancer (NSCLC) cell lines, cyclins D1, D2, and E1 are directly regulated by physiologic concentrations of miR-15a/miR-16. Consistent with these results, overexpression of these miRNAs induces cell cycle arrest in G(1)-G(0). Interestingly, H2009 cells lacking Rb are resistant to miR-15a/miR-16-induced cell cycle arrest, whereas reintroduction of functional Rb resensitizes these cells to miRNA activity. In contrast, down-regulation of Rb in A549 cells by RNA interference confers resistance to these miRNAs. Thus, cell cycle arrest induced by these miRNAs depends on the expression of Rb, confirming that G(1) cyclins are major targets of miR-15a/miR-16 in NSCLC. Our results indicate that miR-15a/miR 16 are implicated in cell cycle control and likely contribute to the tumorigenesis of NSCLC. PMID- 19549911 TI - Spleen tyrosine kinase is overexpressed and represents a potential therapeutic target in chronic lymphocytic leukemia. AB - B-cell receptor signaling contributes to apoptosis resistance in chronic lymphocytic leukemia (CLL), limiting the efficacy of current therapeutic approaches. In this study, we investigated the expression of spleen tyrosine kinase (SYK), a key component of the B-cell receptor signaling pathway, in CLL and its role in apoptosis. Gene expression profiling identified enhanced expression of SYK and downstream pathways in CLL compared with healthy B cells. Immunoblotting showed increased expression and phosphorylation of SYK, PLCgamma(2), signal transducers and activators of transcription 3, and extracellular signal regulated kinase 1/2 in CLL compared with healthy B cells, suggesting enhanced activation of these mediators in CLL. SYK inhibitors reduced phosphorylation of SYK downstream targets and induced apoptosis in primary CLL cells. With respect to prognostic factors, SYK inhibitors exerted stronger cytotoxic effects in unmutated and ZAP70(+) cases. Cytotoxic effects of SYK inhibitors also associated with SYK protein expression, potentially predicting response to therapy. Combination of fludarabine with SYK Inhibitor II or R406 increased cytotoxicity compared with fludarabine therapy alone. We observed no stroma-contact-mediated drug resistance for SYK inhibitors as described for fludarabine treatment. CD40 ligation further enhanced efficacy of SYK inhibition. Our data provide mechanistic insight into the recently observed therapeutic effects of the SYK inhibitor R406 in CLL. Combination of SYK inhibitors with fludarabine might be a novel treatment option particularly for CLL patients with poor prognosis and should be further evaluated in clinical trials. PMID- 19549912 TI - Antitumor activity of gemcitabine and oxaliplatin is augmented by thymoquinone in pancreatic cancer. AB - Previous studies have shown biological activity of thymoquinone, an active compound extracted from Nigella sativa, in pancreatic cancer cells; however, preclinical animal studies are lacking. Here, we report, for the first time, the chemosensitizing effect of thymoquinone to conventional chemotherapeutic agents both in vitro and in vivo using an orthotopic model of pancreatic cancer. In vitro studies revealed that preexposure of cells with thymoquinone (25 mumol/L) for 48 h followed by gemcitabine or oxaliplatin resulted in 60% to 80% growth inhibition compared with 15% to 25% when gemcitabine or oxaliplatin was used alone. Moreover, we found that thymoquinone could potentiate the killing of pancreatic cancer cells induced by chemotherapeutic agents by down-regulation of nuclear factor-kappaB (NF-kappaB), Bcl-2 family, and NF-kappaB-dependent antiapoptotic genes (X-linked inhibitors of apoptosis, survivin, and cyclooxygenase-2). As shown previously by our laboratory, NF-kappaB gets activated on exposure of pancreatic cancer cells to conventional chemotherapeutic agents; interestingly, thymoquinone was able to down-regulate NF-kappaB in vitro, resulting in chemosensitization. In addition to in vitro results, here we show for the first time, that thymoquinone in combination with gemcitabine and/or oxaliplatin is much more effective as an antitumor agent compared with either agent alone. Most importantly, our data also showed that a specific target, such as NF-kappaB, was inactivated in animal tumors pretreated with thymoquinone followed by gemcitabine and/or oxaliplatin. These results provide strong in vivo molecular evidence in support of our hypothesis that thymoquinone could abrogate gemcitabine- or oxaliplatin-induced activation of NF-kappaB, resulting in the chemosensitization of pancreatic tumors to conventional therapeutics. PMID- 19549913 TI - A novel lung metastasis signature links Wnt signaling with cancer cell self renewal and epithelial-mesenchymal transition in basal-like breast cancer. AB - The establishment of metastasis depends on the ability of cancer cells to acquire a migratory phenotype combined with their capacity to recreate a secondary tumor in a distant tissue. In epithelial cancers, such as those of the breast, the epithelial-mesenchymal transition (EMT) is associated with basal-like breast cancers, generates cells with stem-like properties, and enables cancer cell dissemination and metastasis. However, the molecular mechanism(s) that connects stem cell-like characteristics with EMT has yet to be defined. Using an orthotopic model of human breast cancer metastasis to lung, we identified a poor prognosis gene signature, in which several components of the wnt signaling pathway were overexpressed in early lung metastases. The wnt genes identified in this signature were strongly associated with human basal-like breast cancers. We found that inhibiting wnt signaling through LRP6 reduced the capacity of cancer cells to self-renew and seed tumors in vivo. Furthermore, inhibition of wnt signaling resulted in the reexpression of breast epithelial differentiation markers and repression of EMT transcription factors SLUG and TWIST. Collectively, these results provide a molecular link between self-renewal, EMT, and metastasis in basal-like breast cancers. PMID- 19549914 TI - Increased manganese superoxide dismutase expression or treatment with manganese porphyrin potentiates dexamethasone-induced apoptosis in lymphoma cells. AB - Glucocorticoid-induced apoptosis is exploited for the treatment of hematologic malignancies. Innate and acquired resistance limits treatment efficacy; however, resistance mechanisms are not well understood. Previously, using WEHI7.2 murine thymic lymphoma cells, we found that increasing the resistance to hydrogen peroxide (H(2)O(2)) by catalase transfection or selection for H(2)O(2) resistance caused glucocorticoid resistance. This suggests the possibility that increasing H(2)O(2) sensitivity could sensitize the cells to glucocorticoids. In other cell types, increasing manganese superoxide dismutase (MnSOD) can increase intracellular H(2)O(2). The current study showed that increased expression of MnSOD sensitized WEHI7.2 cells to glucocorticoid-induced apoptosis and H(2)O(2). Treatment of WEHI7.2 cells with the catalytic antioxidant Mn(III) meso-tetrakis(N ethylpyridinium-2-yl)porphyrin (MnTE-2-PyP(5+)), a manganoporphyrin, mimicked the effects of increased MnSOD expression. MnTE-2-PyP(5+) also sensitized WEHI7.2 cells to cyclophosphamide and inhibited cell growth; it had no effect on the WEHI7.2 cell response to doxorubicin or vincristine. In primary follicular lymphoma cells, MnTE-2-PyP(5+) increased cell death due to dexamethasone. Treatment of H9c2 cardiomyocytes with MnTE-2-PyP(5+) inhibited doxorubicin cytotoxicity. The profile of MnTE-2-PyP(5+) effects suggests MnTE-2-PyP(5+) has potential for use in hematologic malignancies that are treated with glucocorticoids, cyclophosphamide, and doxorubicin. PMID- 19549915 TI - Nanoscale cellular changes in field carcinogenesis detected by partial wave spectroscopy. AB - Understanding alteration of cell morphology in disease has been hampered by the diffraction-limited resolution of optical microscopy (>200 nm). We recently developed an optical microscopy technique, partial wave spectroscopy (PWS), which is capable of quantifying statistical properties of cell structure at the nanoscale. Here we use PWS to show for the first time the increase in the disorder strength of the nanoscale architecture not only in tumor cells but also in the microscopically normal-appearing cells outside of the tumor. Although genetic and epigenetic alterations have been previously observed in the field of carcinogenesis, these cells were considered morphologically normal. Our data show organ-wide alteration in cell nanoarchitecture. This seems to be a general event in carcinogenesis, which is supported by our data in three types of cancer: colon, pancreatic, and lung. These results have important implications in that PWS can be used as a new method to identify patients harboring malignant or premalignant tumors by interrogating easily accessible tissue sites distant from the location of the lesion. PMID- 19549916 TI - Oncosome formation in prostate cancer: association with a region of frequent chromosomal deletion in metastatic disease. AB - Oncosomes have recently been described as membrane-derived microvesicles secreted by cancer cells, which transfer oncogenic signals and protein complexes across cell boundaries. Here, we show the rapid formation and secretion of oncosomes from DU145 and LNCaP human prostate cancer cells. Oncosome formation was stimulated by epidermal growth factor receptor activation and also by overexpression of membrane-targeted Akt1. Microvesicles shed from prostate cancer cells contained numerous signal transduction proteins and were capable of activating rapid phospho-tyrosine and Akt pathway signaling, and stimulating proliferation and migration, in recipient tumor cells. They also induced a stromal reaction in recipient normal cells. Knockdown of the actin nucleating protein Diaphanous Related Formin 3 (DRF3/Dia2) by RNA interference enhanced rates of oncosome formation, indicating that these structures resemble, and may be identical to, nonapoptotic membrane blebs, a feature of the amoeboid form of cell motility. Analysis of primary and metastatic human prostate tumors using 100K single nucleotide polymorphism arrays revealed a significantly higher frequency of deletion of the locus encoding DRF3 (DIAPH3) in metastatic tumors (P = 0.001) in comparison with organ-confined tumors. Fluorescence in situ hybridization confirmed increased chromosomal loss of DIAPH3 in metastatic tumors in a different cohort of patients (P = 0.006). These data suggest that microvesicles shed from prostate cancer cells can alter the tumor microenvironment in a manner that may promote disease progression. They also show that DRF3 is a physiologically relevant protein that seems to regulate this process. PMID- 19549917 TI - Lymphopenia as a prognostic factor for overall survival in advanced carcinomas, sarcomas, and lymphomas. AB - Lymphopenia is frequent in advanced cancers and predicts the toxicity of chemotherapy. Its effect on relapse and survival is uncertain. Its prognostic value for survival was analyzed in three databases of previously reported prospective multicenter studies: (a) FEC chemotherapy in metastatic breast carcinoma; (b) CYVADIC in advanced soft tissue sarcoma (European Organization for Research and Treatment of Cancer-Soft Tissue and Bone Sarcoma Group 62791); and (c) prospective, consecutive phase III studies of aggressive diffuse large-cell non-Hodgkin's lymphomas conducted at Centre Leon Berard between 1987 and 1993. Univariate and multivariate analyses of prognostic factors for survival were performed. The incidence of lymphopenia of <1,000/microL before treatment was constant among the series: 25%, 24%, and 27%, respectively. Lymphopenia was significantly more frequent (P < 0.05) in metastatic breast cancer patients with performance status (PS) of >1, non-Hodgkin's lymphoma patients with international prognostic index (IPI) of > 0, and advanced soft tissue sarcoma and metastatic breast cancer patients with bone metastases. Inunivariate analysis, lymphopenia of <1,000/microL significantly correlated to overall survival in patients with metastatic breast cancer (median, 10 versus 14 mo; P < 0.0001), advanced soft tissue sarcoma (median, 5 versus 10 months; P < 0.01), and non-Hodgkin lymphoma (median, 11 versus 94 months; P < 0.0001). In multivariate analysis (Cox model), lymphopenia was an independent prognostic factor for overall survival in metastatic breast cancer [RR (relative risk), 1.8; 95% CI (confidence interval), 1.3-2.4] along with liver metastases and PS; in advanced soft tissue sarcoma (RR, 1.46; 95% CI, 1.0-2.1) along with liver metastases, lung metastases, and PS; and in non-Hodgkin's lymphoma (RR, 1.48; 95% CI, 1.03-2.1) along with IPI. Our findings show that lymphopenia is an independent prognostic factor for overall and progression-free survival in several cancers. PMID- 19549918 TI - Active Notch1 confers a transformed phenotype to primary human melanocytes. AB - The importance of mitogen-activated protein kinase signaling in melanoma is underscored by the prevalence of activating mutations in N-Ras and B-Raf, yet clinical development of inhibitors of this pathway has been largely ineffective, suggesting that alternative oncogenes may also promote melanoma. Notch is an interesting candidate that has only been correlated with melanoma development and progression; a thorough assessment of tumor-initiating effects of activated Notch on human melanocytes would clarify the mounting correlative evidence and perhaps identify a novel target for an otherwise untreatable disease. Analysis of a substantial panel of cell lines and patient lesions showed that Notch activity is significantly higher in melanomas than their nontransformed counterparts. The use of a constitutively active, truncated Notch transgene construct (N(IC)) was exploited to determine if Notch activation is a "driving" event in melanocytic transformation or instead a "passenger" event associated with melanoma progression. N(IC)-infected melanocytes displayed increased proliferative capacity and biological features more reminiscent of melanoma, such as dysregulated cell adhesion and migration. Gene expression analyses supported these observations and aided in the identification of MCAM, an adhesion molecule associated with acquisition of the malignant phenotype, as a direct target of Notch transactivation. N(IC)-positive melanocytes grew at clonal density, proliferated in limiting media conditions, and also exhibited anchorage independent growth, suggesting that Notch alone is a transforming oncogene in human melanocytes, a phenomenon not previously described for any melanoma oncogene. This new information yields valuable insight into the basic epidemiology of melanoma and launches a realm of possibilities for drug intervention in this deadly disease. PMID- 19549920 TI - Insulin-like growth factor-II methylation status in lymphocyte DNA and colon cancer risk in the Northern Sweden Health and Disease cohort. AB - Loss of imprinting (LOI) of the insulin-like growth factor II (IGFII) gene is a frequent phenomenon in colorectal tumor tissues. Previous reports indicated that subjects with colorectal neoplasias show LOI of IGFII in circulating lymphocytes. Furthermore, LOI of IGFII is strongly related to the methylation of a differentially methylated region (DMR) in intron 2 of IGFII, suggesting that the methylation status could serve as a biomarker for early detection. Thus, hypermethylation of this DMR, even at a systemic level, e.g., in lymphocyte DNA, could be used for screening for colon cancer. To validate this, we performed a case-control study of 97 colon cancer cases and 190 age-matched and gender matched controls, nested within the prospective Northern Sweden Health and Disease Study cohort. Methylation levels of the IGFII-DMR in lymphocyte DNA were measured at two specific CpG sites of the IGFII-DMR using a mass-spectrometric method called short oligonucleotide mass analysis, the measurements of which showed high reproducibility between replicate measurements for the two CpG sites combined and showed almost perfect validity when performed on variable mixtures of methylated and unmethylated standards. Mean fractions of CpG methylation, for the two CpG sites combined, were identical for cases and controls (0.47 and 0.46, respectively; P(difference) = 0.75), and logistic regression analyses showed no relationship between colon cancer risk and quartile levels of CpG methylation. The results from this study population do not support the hypothesis that colon cancer can be predicted from the different degrees of methylation of DMR in the IGFII gene from lymphocyte DNA. PMID- 19549919 TI - Breast cancer migration and invasion depend on proteasome degradation of regulator of G-protein signaling 4. AB - Aberrant signaling through G-protein coupled receptors promotes metastasis, the major cause of breast cancer death. We identified regulator of G-protein signaling 4 (RGS4) as a novel suppressor of breast cancer migration and invasion, important steps of metastatic cascades. By blocking signals initiated through G(i)-coupled receptors, such as protease-activated receptor 1 and CXC chemokine receptor 4, RGS4 disrupted Rac1-dependent lamellipodia formation, a key step involved in cancer migration and invasion. RGS4 has GTPase-activating protein (GAP) activity, which inhibits G-protein coupled receptor signaling by deactivating G-proteins. An RGS4 GAP-deficient mutant failed to inhibit migration and invasion of breast cancer cells in both in vitro assays and a mouse xenograft model. Interestingly, both established breast cancer cell lines and human breast cancer specimens showed that the highest levels of RGS4 protein were expressed in normal breast epithelia and that RGS4 down-regulation by proteasome degradation is an index of breast cancer invasiveness. Proteasome blockade increased endogenous RGS4 protein to levels that markedly inhibit breast cancer cell migration and invasion, which was reversed by an RGS4-targeted short hairpin RNA. Our findings point to the existence of a mechanism for posttranslational regulation of RGS4 function, which may have important implications for the acquisition of a metastatic phenotype by breast cancer cells. Preventing degradation of RGS4 protein should attenuate aberrant signal inputs from multiple G(i)-coupled receptors, thereby retarding the spread of breast cancer cells and making them targets for surgery, radiation, and immune treatment. PMID- 19549921 TI - ADAM23 negatively modulates alpha(v)beta(3) integrin activation during metastasis. AB - The ADAM23 gene is frequently silenced in different types of tumors, and, in breast tumors, silencing is correlated with tumor progression, suggesting that it might be associated with the acquisition of a metastatic phenotype. ADAM23 exerts its function mainly through the disintegrin domain, because its metalloprotease domain is inactive. Analysis of ADAM23 binding to integrins has revealed a specific interaction with alpha(v)beta(3) integrin mediated by the disintegrin domain. Altered expression of alpha(v)beta(3) integrin has been observed in different types of tumors, and expression of this integrin in the activated form has been shown to promote metastasis formation. Here, we investigated the possibility that interaction between ADAM23 and alpha(v)beta(3) integrin might negatively modulate alpha(v)beta(3) activation during metastatic progression. ADAM23 expression was knocked down using short hairpin RNA in the MDA-MB-435 cell line, which has been extensively used as a model for alpha(v)beta(3) integrin activation. Ablation of ADAM23 enhanced alpha(v)beta(3) integrin activation by at least 2- to 4-fold and ADAM23 knockdown cells showed enhanced migration and adhesion to classic alpha(v)beta(3) integrin ligands. Ablation of ADAM23 expression also enhanced pulmonary tumor cell arrest in immunodeficient mice. To complement our findings with clinical evidence, we showed that silencing of ADAM23 gene by DNA promoter hypermethylation in a collection of 94 primary breast tumors was significantly associated with lower distant metastases-free and disease-specific survivals and was an independent prognostic factor for poor disease outcome. Our results strongly support a functional role of ADAM23 during metastatic progression by negatively modulating alpha(v)beta(3) integrin activation. PMID- 19549923 TI - Assessment of sensory function in the National Social Life, Health, and Aging Project. AB - OBJECTIVES: The National Social Life, Health, and Aging Project assessed functioning of all 5 senses using both self-report and objective measures. We evaluate the performance of the objective measures and model differences in sensory function by gender and age. In the process, we demonstrate how to use and interpret these measures. METHODS: Distance vision was assessed using a standard Sloan eye chart, and touch was measured using a stationary 2-point discrimination test applied to the index fingertip of the dominant hand. Olfactory function (both intensity detection and odor identification) was assessed using odorants administered via felt-tip pens. Gustatory function was measured via identification of four taste strips. RESULTS: The performance of the objective measures was similar to that reported for previous studies, as was the relationship between sensory function and both gender and age. DISCUSSION: Sensory function is important in studies of aging and health both because it is an important health outcome and also because a decline in functioning can be symptomatic of or predict other health conditions. Although the objective measures provide considerably more precision than the self-report items, the latter can be valuable for imputation of missing data and for understanding differences in how older adults perceive their own sensory ability. PMID- 19549924 TI - Heparan sulfate promotes the aggregation of HDL-associated serum amyloid A: evidence for a proamyloidogenic histidine molecular switch. AB - During inflammatory diseases, serum amyloid A (SAA), an acute-phase apolipoprotein of HDL, can assemble into tissue deposits called AA amyloids. The mechanism and physiological factors promoting amyloidosis are largely unknown but likely involve heparan sulfate (HS), a glycosaminoglycan colocalized with all types of amyloids. In this study, we explored HDL-SAA:HS interactions using in vitro and cell culture assays to identify HS-binding domains that promote the conversion of native SAA into AA amyloid. HS causes the remodeling of HDL-SAA at mildly acidic pH, producing SAA-rich aggregates. A sequence motif in SAA responsible for this conversion was identified that contains a pH-sensitive heparin/HS-binding site, functions as a ligand for a cell surface receptor, and acts as a structural focal point for SAA aggregation. Synthetic peptides corresponding to this region promoted the deposition of AA amyloid in a monocyte culture model for AA amyloidogenesis. The effects were peptide sequence specific and reliant on the protonation of H36. We conclude that a highly conserved motif required for SAA binding to macrophages can, under acidic pH conditions and in an HS-dependent manner, also act as a molecular switch, directing SAA misfolding into AA amyloid. Similar histidine-dependent HS-binding sites are also found in other amyloidogenic polypeptides. PMID- 19549922 TI - Stimulating the GPR30 estrogen receptor with a novel tamoxifen analogue activates SF-1 and promotes endometrial cell proliferation. AB - Estrogens and selective estrogen receptor (ER) modulators such as tamoxifen are known to increase uterine cell proliferation. Mounting evidence suggests that estrogen signaling is mediated not only by ERalpha and ERbeta nuclear receptors, but also by GPR30 (GPER), a seven transmembrane (7TM) receptor. Here, we report that primary human endometriotic H-38 cells express high levels of GPR30 with no detectable ERalpha or ERbeta. Using a novel tamoxifen analogue, STX, which activates GPR30 but not ERs, significant stimulation of the phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathways was observed in H-38 cells and in Ishikawa endometrial cancer cells expressing GPR30; a similar effect was observed in JEG3 choriocarcinoma cells. STX treatment also increased cellular pools of phosphatidylinositol (3,4,5) triphosphate, a proposed ligand for the nuclear hormone receptor SF-1 (NR5A1). Consistent with these findings, STX, tamoxifen, and the phytoestrogen genistein were able to increase SF-1 transcription, promote Ishikawa cell proliferation, and induce the SF-1 target gene aromatase in a GPR30-dependent manner. Our findings suggest a novel signaling paradigm that is initiated by estrogen activation of the 7TM receptor GPR30, with signal transduction cascades (PI3K and MAPK) converging on nuclear hormone receptors (SF-1/LRH-1) to modulate their transcriptional output. We propose that this novel GPR30/SF-1 pathway increases local concentrations of estrogen, and together with classic ER signaling, mediate the proliferative effects of synthetic estrogens such as tamoxifen, in promoting endometriosis and endometrial cancers. PMID- 19549925 TI - Sixty years of Pharmacological Reviews: has the role of review articles in biomedical sciences changed and, if so, how does this affect Pharmacological Reviews? PMID- 19549926 TI - Orexin/hypocretin: a neuropeptide at the interface of sleep, energy homeostasis, and reward system. AB - Recent studies have implicated the orexin system as a critical regulator of sleep/wake states as well as feeding behavior and reward processes. Orexin deficiency results in narcolepsy in humans, dogs, and rodents, suggesting that the orexin system is particularly important for maintenance of wakefulness. In addition, orexin deficiency also cause abnormalities in energy homeostasis and reward systems. Orexin activates waking active monoaminergic and cholinergic neurons in the hypothalamus and brainstem regions to maintain a long, consolidated waking period. Orexin neurons receive abundant input from the limbic system. Orexin neurons also have reciprocal links with the hypothalamic arcuate nucleus, which regulates feeding. Moreover, the responsiveness of orexin neurons to peripheral metabolic cues, such as leptin and glucose, suggest that these neurons have important role as a link between the energy homeostasis and vigilance states. Orexin neurons also have a link with the dopaminergic reward system in the ventral tegmental nucleus. These findings suggest that the orexin system interacts with systems that regulate emotion, reward, and energy homeostasis to maintain proper vigilance states. Therefore, this system may be a potentially important therapeutic target for treatment of sleep disorder, obesity, emotional stress, and addiction. PMID- 19549928 TI - Lutein accumulation in the absence of zeaxanthin restores nonphotochemical quenching in the Arabidopsis thaliana npq1 mutant. AB - Plants protect themselves from excess absorbed light energy through thermal dissipation, which is measured as nonphotochemical quenching of chlorophyll fluorescence (NPQ). The major component of NPQ, qE, is induced by high transthylakoid DeltapH in excess light and depends on the xanthophyll cycle, in which violaxanthin and antheraxanthin are deepoxidized to form zeaxanthin. To investigate the xanthophyll dependence of qE, we identified suppressor of zeaxanthin-less1 (szl1) as a suppressor of the Arabidopsis thaliana npq1 mutant, which lacks zeaxanthin. szl1 npq1 plants have a partially restored qE but lack zeaxanthin and have low levels of violaxanthin, antheraxanthin, and neoxanthin. However, they accumulate more lutein and alpha-carotene than the wild type. szl1 contains a point mutation in the lycopene beta-cyclase (LCYB) gene. Based on the pigment analysis, LCYB appears to be the major lycopene beta-cyclase and is not involved in neoxanthin synthesis. The Lhcb4 (CP29) and Lhcb5 (CP26) protein levels are reduced by 50% in szl1 npq1 relative to the wild type, whereas other Lhcb proteins are present at wild-type levels. Analysis of carotenoid radical cation formation and leaf absorbance changes strongly suggest that the higher amount of lutein substitutes for zeaxanthin in qE, implying a direct role in qE, as well as a mechanism that is weakly sensitive to carotenoid structural properties. PMID- 19549929 TI - Qualification of cardiac troponin I concentration in mouse serum using isoproterenol and implementation in pharmacology studies to accelerate drug development. AB - Cardiac troponin I is a useful biomarker of myocardial injury, but its use in mice and application to early drug discovery are not well described. The authors investigated the relationship between cTnI concentration in serum and histologic lesions in heart tissue from mice treated with isoproterenol (ISO). Cardiac TnI concentrations in serum increased in a dose-dependant manner and remained increased twenty-four to forty-eight hours after a single administration of isoproterenol. Increased cTnI concentration was of greater magnitude and longer duration than increased fatty acid binding protein 3 concentration, aspartate aminotransferase activity, and creatine kinase activity in serum. Isoproterenol induced increases in cTnI concentrations were both greater and more sustained in BALB/c than in CD1 mice and correlated with incidence and severity of lesions observed in heart sections from both strains. In drug development studies in BALB/c mice with novel kinase inhibitors, cTnI concentration was a reliable stand alone biomarker of cardiac injury and was used in combination with measurements of in vivo target inhibition to demonstrate an off-target contribution to cardiotoxicity. Additional attributes, including low cost and rapid turnaround time, made cTnI concentration in serum invaluable for detecting cardiotoxicity, exploring structure-activity relationships, and prioritizing development of compounds with improved safety profiles early in drug discovery. PMID- 19549931 TI - Microbial removal rates in subsurface media estimated from published studies of field experiments and large intact soil cores. AB - Information about the microbial removal efficiencies of subsurface media is essential for assessing the risk of water contamination, estimating setback distances between disposal fields and receiving waters, and selecting suitable sites for wastewater reclamation. By analyzing published data from field experiments and large intact soil cores, an extensive database of microbial removal rates was established for a wide range of subsurface media. High microbial removal rates were found in volcanic soils, pumice sand, fine sand, and highly weathered aquifer rocks. Low removal rates were found in structured clayey soils, stony soils, coarse gravel aquifers, fractured rocks, and karst limestones. Removal rates were lower for enteroviruses than for other human viruses; for MS2 phage than for other phage species; for waste-associated microbes than for those cultivated in the laboratory; and for contaminated media than for uncontaminated media. Microbial removal rates are inversely correlated with infiltration rates and transport velocity. The assumption of first-order law, or a constant removal rate (when the transport scale reaches a representative elementary volume), is appropriate for most of field data analyzed. However 30% of the datasets (26 out of 87 pairs) are better described with the power law, implying reduced removal rates with transport distance. The latter is most prominent for organically contaminated media, especially in relatively fine aquifer media. The presence of organic matter, heterogeneity in microbial properties, change in solution chemistry, detachment, and physical straining, may have caused the discrepancies from the first-order law traditionally used in transport models for describing microbial removal. PMID- 19549930 TI - Emodin has cytotoxic and protective effects in rat C6 glioma cells: roles of Mdr1a and nuclear factor kappaB in cell survival. AB - 1,3,8-Trihydroxy-6-methylanthaquinone (emodin) is recognized as an antiproliferative compound. In the present study, however, we show that emodin has both toxic and survival effects in glioma cells and that the survival effects involve Mdr1a. Emodin inhibited the proliferation and induced apoptosis of C6 cells in a 12-h treatment, but C6 cells survived a 72-h drug treatment, indicating resistance to emodin. Emodin-induced apoptosis was reduced by inhibition of the expression and activation of apoptosis-associated proteins including p53, Bax, Bcl-2, Fas, and caspase-3. C6 cells could express antioxidant proteins (superoxide dismutase and catalase) to decrease reactive oxygen species induced cytotoxicity of emodin and overexpress multidrug resistance genes (Mdr1a, MRP2, MRP3, and MRP6) to decrease the intracellular accumulation of emodin. Electrophoretic mobility shift analysis showed that emodin decreased nuclear factor kappaB (NF-kappaB) expression in 24 h of treatment, but in 48 h, emodin increased NF-kappaB activity. A confocal microscope showed that emodin induced NF kappaB translocation from cytoplasm to nuclei. C6 cells would activate the mitogen-activated protein kinase survival pathway and express the DNA repair gene (MGMT) and associated proteins (PARP and XRCC1) to recover the cell activity. C6 cells also expressed GRP78 to decrease emodin-induced endoplasmic reticulum (ER) stress that would cause apoptosis in C6 cells, and GRP78 inhibited the expression of GADD153 to enhance the expression of Bcl-2 that could balance the ER- and mitochondria-induced apoptosis of C6 cells. PMID- 19549932 TI - Pilot scale facility to determine gaseous emissions from livestock slurry during storage. AB - Livestock production is a growing source of air pollution, locally and to the wider environment. Improved livestock manure management has the potential to reduce environmental impacts, but there is a need for methodologies to precisely quantify emissions. This paper describes and evaluates a novel storage facility for livestock slurry consisting of eight 6.5-m(3) cylindrical units. The stores may be equipped with airtight covers and ventilated during storage or during measurement only. Each store has eight air inlets (160 mm diameter) and a single outlet in the cover connected to a main ventilation duct. The stores can also be used as static enclosures. Ventilation can be regulated within the range of 50 to 250 m(3) h(-1). A gas sampling line enables sampling of odorants using automatic thermal desorption tubes, ammonia using acid traps, and greenhouse gases using gas sampling bags (pooled samples) or a syringe (time point samples). Complete recovery of CH(4) independent of ventilation rate was demonstrated. Vertical profiles of CO(2) and CH(4) above the slurry surface with and without ventilation and mixing of the headspace indicated methane oxidation activity in the surface crust. p-Cresol and 4-ethyl phenol emission from pig slurry was identified by GC MS analysis of odor collected on adsorption tubes. Ammonia emissions between 0 and 166 mg N m(-2) h(-1) were observed during storage of pig slurry with and without surface crust and cover. A comparison of pooled and averaged time point measurements of CO(2), CH(4), and N(2)O indicated that pooled samples account for the diurnal variations under realistic storage conditions. PMID- 19549933 TI - Fertilizer application timing influences greenhouse gas fluxes over a growing season. AB - Microbial production and consumption of greenhouse gases (GHG) is influenced by temperature and nutrients, especially during the first few weeks after agricultural fertilization. The effect of fertilization on GHG fluxes should occur during and shortly after application, yet data indicating how application timing affects both GHG fluxes and crop yields during a growing season are lacking. We designed a replicated (n = 5) field experiment to test for the short term effect of fertilizer application timing on fluxes of methane (CH(4)), carbon dioxide (CO(2)), and nitrous oxide (N(2)O) over a growing season in the northern Great Plains. Each 0.30-ha plot was planted to maize (Zea mays L.) and treated similarly with the exception of fertilizer timing: five plots were fertilized with urea in early spring (1 April) and five plots were fertilized with urea in late spring (13 May). We hypothesized time-integrated fluxes over a growing season would be greater for the late-spring treatment, resulting in a greater net GHG flux, as compared to the early-spring treatment. Data collected on 59 dates and integrated over a 5-mo time course indicated CO(2) fluxes were greater (P < 0.0001) and CH(4) fluxes were lower (P < 0.05) for soils fertilized in late spring. Net GHG flux was also significantly affected by treatment, with 0.84 +/- 0.11 kg CO(2) equivalents m(-2) for early spring and 1.04 +/- 0.13 kg CO(2) equivalents m(-2) for late spring. Nitrous oxide fluxes, however, were similar for both treatments. Results indicate fertilizer application timing influences net GHG emissions in dryland cropping systems. PMID- 19549934 TI - Recreating a functioning forest soil in reclaimed oil sands in northern alberta: an approach for measuring success in ecological restoration. AB - During oil-sands mining all vegetation, soil, overburden, and oil sand is removed, leaving pits several kilometers wide and up to 100 m deep. These pits are reclaimed through a variety of treatments using subsoil or a mixed peat mineral soil cap. Using nonmetric multidimensional scaling and cluster analysis of measurements of ecosystem function, reclamation treatments of several age classes were compared with a range of natural forest ecotypes to discover which treatments had created ecosystems similar to natural forest ecotypes and at what age this occurred. Ecosystem function was estimated from bioavailable nutrients, plant community composition, litter decomposition rate, and development of a surface organic layer. On the reclamation treatments, availability of nitrate, calcium, magnesium, and sulfur were generally higher than in the natural forest ecotypes, while ammonium, P, K, and Mn were generally lower. Reclamation treatments tended to have more bare ground, grasses, and forbs but less moss, lichen, shrubs, trees, or woody debris than natural forests. Rates of litter decomposition were lower on all reclamation treatments. Development of an organic layer appeared to be facilitated by the presence of shrubs. With repeated applications of fertilizers, measured variables for the peat-mineral amendments fell within the range of natural variability at about 20 yr. An intermediate subsoil layer reduced the need for fertilizer and conditions resembling natural forests were reached about 15 yr after a single fertilizer application. Treatments over tailings sand receiving only one application of fertilizer appeared to be on a different trajectory to a novel ecosystem. PMID- 19549935 TI - Simultaneous removal of polycyclic aromatic hydrocarbons and copper from soils using ethyl lactate-amended EDDS solution. AB - A new approach using aqueous ethyl lactate-modified [S,S] ethylenediaminedisuccinic acid (EDDS) washing solutions was examined in the laboratory for the simultaneous removal of phenanthrene, pyrene, and Cu from contaminated soils. Ethyl lactate demonstrated greater solubilization efficiency for phenanthrene and pyrene than ethanol. Thus ethyl lactate has a great potential for extracting polycyclic aromatic hydrocarbons (PAHs) from contaminated soils. For soils with varying properties, removal efficiencies were found to be negatively correlated with soil organic carbon contents. Aqueous EDDS solution effectively extracted Cu from soils. The extraction efficiency reached about 36.7% at a EDDS:Cu molar ratio of 5. The addition of ethyl lactate in EDDS solution (EDDS/Cu molar ratio = 2) efficiently enhanced the extraction of the PAHs and also significantly increased the Cu removal from 34.8 to 42.9%. The latter was mainly attributed to the fact that ethyl lactate increases the stability constant for Cu-EDDS complexes, hence shifting the degree of desorption of Cu from soil. Sequential extraction indicated that the EDDS/ethyl lactate solution extracted Cu primarily from the acid extractable fraction, the reducible fraction and the oxidizable fraction. The results suggest that simultaneous removal of PAHs and heavy metals from contaminated soils is feasible by soil washing using an aqueous EDDS solution enhanced with ethyl lactate. PMID- 19549936 TI - The role of indigenous microorganisms in the biodegradation of naturally occurring petroleum, the reduction of iron, and the mobilization of arsenite from west bengal aquifer sediments. AB - High levels of naturally occurring arsenic are found in the shallow reducing aquifers of West Bengal, Bangladesh, and other areas of Southeast Asia. These aquifers are used extensively for drinking water and irrigation by the local population. Mechanisms for its release are unclear, although increasing evidence points to a microbial control. The type of organic matter present is of vital importance because it has a direct impact on the rate of microbial activity and on the amount of arsenic released into the ground water. The discovery of naturally occurring hydrocarbons in an arsenic-rich aquifer from West Bengal provides a source of potential electron donors for this process. Using microcosm based techniques, seven sediments from a site containing naturally occurring hydrocarbons in West Bengal were incubated with synthetic ground water for 28 d under anaerobic conditions without the addition of an external electron donor. Arsenic release and Fe(III) reduction appeared to be microbially mediated, with variable rates of arsenic mobilization in comparison to Fe(III) reduction, suggesting that multiple processes are involved. All sediments showed a preferential loss of petroleum-sourced n-alkanes over terrestrially sourced sedimentary hydrocarbons n-alkanes during the incubation, implying that the use of petroleum-sourced n-alkanes could support, directly or indirectly, microbial Fe(III) reduction. Samples undergoing maximal release of As(III) contained a significant population of Sulfurospirillum sp., a known As(V)-reducing bacterium, providing the first evidence that such organisms may mediate arsenic release from West Bengali aquifers. PMID- 19549937 TI - Mercury adsorption-desorption and transport in soils. AB - Kinetic sorption and column miscible displacement transport experiments were performed to quantify the extent of retention/release and the mobility of mercury in different soils. Results indicated that adsorption of mercury was rapid and highly nonlinear with sorption capacities having the following sequence: Sharkey clay > Olivier loam > Windsor sand. Mercury adsorption by all soils was strongly irreversible where the amounts released or desorbed were often less than 1% of that applied. Moreover, the removal of soil organic matter resulted in a decrease of mercury adsorption in all soils. Adsorption was described with limited success using a nonlinear (Freundlich) model. Results from the transport experiments indicated that the mobility of mercury was highly retarded, with extremely low concentrations of mercury in column effluents. Furthermore, mercury breakthrough curves exhibited erratic patterns with ill-distinguished peaks. Therefore, mercury is best regarded as strongly retained and highly "immobile" in the soils investigated. This is most likely due to highly stable complex formation (irreversible forms) and strong binding to high-affinity sites. In a column packed with reference sand material, a symmetric breakthrough curve was obtained where the recovery of mercury in the leachate was only 17.3% of that applied. Mercury retention by the reference sand was likely due to adsorption by quartz and metal-oxides. PMID- 19549938 TI - Heavy metal contamination from electronic waste recycling at Guiyu, Southeastern China. AB - This study evaluated the effect of electronic waste (e-waste) recycling activities at a regional center, Guiyu, Guangdong, China on heavy metal pollution in the surrounding waterway system. Surface water and sediment collected from the Lianjiang River and its tributaries were analyzed for concentrations of Cu, Pb, Ni, Cd, Hg, and As. Spatially, heavy metals in water and sediment samples decreased downstream of Lianjiang River from Guiyu to the estuary at Haimen Bay. Temporally, heavy metal concentrations in the surface water were high in the rainy season and low in the dry season. In particular, concentrations of Cu from surface water in the Guiyu area were 2.4 to 131 times the reference background concentration, followed in descending order by Ni, Cd, Pb, Hg, and As. However, when compared with China's Environmental Quality Standard, Cd was considered the most harmful element, followed by Hg, Pb, and Cu. Concentrations of Cu in sediment samples in the Guiyu area were 3.2 to 429 times the reference background concentration, followed by Ni, Hg, Pb, Cd, and As. Our findings suggested that high concentrations of heavy metal observed in Guiyu, Lianjiang River and the Haimen Bay could be attributed to the direct effects of e-waste recycling in Guiyu. PMID- 19549939 TI - Effectiveness and efficacy of conservation options after potato harvest. AB - Soil erosion and phosphorus (P) runoff can be severe in potato production systems in the Northeast USA, which are characterized by intensive tillage, minimal ground cover, low crop residue return, and steep slopes. We used rainfall simulators in the greenhouse and field to assess sediment and P movement associated with two conservation practices: straw mulching and application of polyacrylamide (PAM). In the greenhouse, a Nokomis sandy loam soil (fine-loamy, mixed, frigid Typic Haplorthods) was packed into 0.2 by 1.0 m boxes and subjected to four rainfall events at an intensity of 70 mm h(-1). Runoff amount, sediment concentration, and inorganic and sediment-bound P were measured for 30 min after initiation of runoff. Linear increases in straw mulch biomass (up to equivalent of 3000 kg ha(-1)) resulted in exponential decreases in sediment and P loss. Mulch applied at rates as low as 600 kg ha(-1) provided nearly 50% ground cover and reduced sediment movement and sediment-bound P concentration and loss by >50%. Higher application rates reduced sediment loss by up to 95% but contributed dissolved reactive P (DRP) to runoff water. Field observations using simulated rainfall on mulch-covered and bare soil were consistent with greenhouse results. Linear increases in PAM application rate (to 20 kg ha(-1)) also reduced sediment loss. The efficacy of this practice decreases slightly with successive rainfall events but still had significant benefit through four simulated rainfalls on soil packed into boxes. This was not the case in the field where the effect of PAM was limited to the first two rainfall events. In general, runoff volume was not strongly influenced by any of these practices, and most of the P loss was comprised of sediment-bound P. Both conservation practices are effective at reducing soil and nutrient loss in intensive potato systems. PMID- 19549940 TI - Uncertainty evaluation of coliform bacteria removal from vegetated filter strip under overland flow condition. AB - Vegetated filter strips (VFS) have become an important component of water quality improvement by reducing sediment and nutrients transport to surface water. This management practice is also beneficial for controlling manure-borne pathogen transport to surface water. The objective of this work was to assess the VFS efficiency and evaluate the uncertainty in predicting the microbial pollutant removal from overland flow in VFS. We used the kinematic wave overland flow model as implemented in KINEROS2 coupled with the convective-dispersive overland transport model which accounts for the reversible attachment-detachment and surface straining of infiltrating bacteria. The model was successfully calibrated with experimental data obtained from a series of simulated rainfall experiments at vegetated and bare sandy loam and clay loam plots, where fecal coliforms were released from manure slurry applied on the top of the plots. The calibrated model was then used to assess the sensitivity of the VFS efficiency to the model parameters, rainfall duration, and intensity for a case study with a 6-m VFS placed at the edge of 200-m long field. The Monte Carlo simulations were also performed to evaluate the uncertainty associated with the VFS efficiency given the uncertainty in the model parameters and key inputs. The VFS efficiency was found to be <95% in 25%, <75% in 23%, and <25% in 20% of cases. Relatively long high-intensity rainfalls, low hydraulic conductivities, low net capillary drives of soil, and high soil moisture contents before rainfalls caused the partial failure of VFS to retain coliforms from the infiltration excess runoff. PMID- 19549941 TI - Estimating phosphorus loss in runoff from manure and fertilizer for a phosphorus loss quantification tool. AB - Nonpoint-source pollution of fresh waters by P is a concern because it contributes to accelerated eutrophication. Given the state of the science concerning agricultural P transport, a simple tool to quantify annual, field scale P loss is a realistic goal. We developed new methods to predict annual dissolved P loss in runoff from surface-applied manures and fertilizers and validated the methods with data from 21 published field studies. We incorporated these manure and fertilizer P runoff loss methods into an annual, field-scale P loss quantification tool that estimates dissolved and particulate P loss in runoff from soil, manure, fertilizer, and eroded sediment. We validated the P loss tool using independent data from 28 studies that monitored P loss in runoff from a variety of agricultural land uses for at least 1 yr. Results demonstrated (i) that our new methods to estimate P loss from surface manure and fertilizer are an improvement over methods used in existing Indexes, and (ii) that it was possible to reliably quantify annual dissolved, sediment, and total P loss in runoff using relatively simple methods and readily available inputs. Thus, a P loss quantification tool that does not require greater degrees of complexity or input data than existing P Indexes could accurately predict P loss across a variety of management and fertilization practices, soil types, climates, and geographic locations. However, estimates of runoff and erosion are still needed that are accurate to a level appropriate for the intended use of the quantification tool. PMID- 19549942 TI - Evaluating nonpoint source critical source area contributions at the watershed scale. AB - Areas with disproportionately high pollutant losses (i.e., critical source areas [CSAs]) have been widely recognized as priority areas for the control of nonpoint source pollution. The identification and evaluation of CSAs at the watershed scale allows state and federal programs to implement soil and water conservation measures where they are needed most. Despite many potential advantages, many state and federal conservation programs do not actively target CSAs. There is a lack of research identifying the total CSA pollutant contribution at the watershed scale, and there is no quantitative assessment of program effectiveness if CSAs are actively targeted. The purpose of this research was to identify and quantify sediment and total phosphorus loads originating from CSAs at the watershed scale using the Soil and Water Assessment Tool. This research is a synthesis of CSA targeting studies performed in six Oklahoma priority watersheds from 2001 to 2007 to aid the Oklahoma Conservation Commission in the prioritized placement of subsidized conservation measures. Within these six watersheds, 5% of the land area yielded 50% of sediment and 34% of the phosphorus load. In watersheds dominated by agriculture, the worst 5% of agricultural land contributed, on average, 22% of the total agricultural pollutant load. Pollutant loads from these agricultural CSAs were more than four times greater than the average load from agricultural areas within the watershed. Conservation practices implemented in these areas can be more effective because they have the opportunity to treat more pollutant. The evaluation of CSAs and prioritized implementation of conservation measures at the watershed scale has the potential to significantly improve the effectiveness of state and federally sponsored water quality programs. PMID- 19549927 TI - Extracellular matrix molecules: potential targets in pharmacotherapy. AB - The extracellular matrix (ECM) consists of numerous macromolecules classified traditionally into collagens, elastin, and microfibrillar proteins, proteoglycans including hyaluronan, and noncollagenous glycoproteins. In addition to being necessary structural components, ECM molecules exhibit important functional roles in the control of key cellular events such as adhesion, migration, proliferation, differentiation, and survival. Any structural inherited or acquired defect and/or metabolic disturbance in the ECM may cause cellular and tissue alterations that can lead to the development or progression of disease. Consequently, ECM molecules are important targets for pharmacotherapy. Specific agents that prevent the excess accumulation of ECM molecules in the vascular system, liver, kidney, skin, and lung; alternatively, agents that inhibit the degradation of the ECM in degenerative diseases such as osteoarthritis would be clinically beneficial. Unfortunately, until recently, the ECM in drug discovery has been largely ignored. However, several of today's drugs that act on various primary targets affect the ECM as a byproduct of the drugs' actions, and this activity may in part be beneficial to the drugs' disease-modifying properties. In the future, agents and compounds targeting directly the ECM will significantly advance the treatment of various human diseases, even those for which efficient therapies are not yet available. PMID- 19549943 TI - Denitrification in wood chip bioreactors at different water flows. AB - Subsurface drainage in agricultural watersheds exports a large quantity of nitrate-nitrogen (NO(3)-N) and concentrations frequently exceed 10 mg L(-1). A laboratory column study was conducted to investigate the ability of a wood chip bioreactor to promote denitrification under mean water flow rates of 2.9, 6.6, 8.7 and 13.6 cm d(-1) which are representative of flows entering subsurface drainage tiles. Columns were packed with wood chips and inoculated with a small amount of oxidized till and incubated at 10 degrees C. Silicone sampling cells at the effluent ports were used for N(2)O sampling. (15)Nitrate was added to dosing water at 50 mg L(-1) and effluent was collected and analyzed for NO(3)-N, NH(4) N, and dissolved organic carbon. Mean NO(3)-N concentrations in the effluent were 0.0, 18.5, 24.2, and 35.3 mg L(-1) for the flow rates 2.9, 6.6, 8.7, and 13.6 cm d(-1), respectively, which correspond to 100, 64, 52, and 30% efficiency of removal. The NO(3)-N removal rates per gram of wood increased with increasing flow rates. Denitrification was found to be the dominant NO(3)-N removal mechanism as immobilization of (15)NO(3)-N was negligible compared with the quantity of (15)NO(3)-N removed. Nitrous oxide production from the columns ranged from 0.003 to 0.028% of the N denitrified, indicating that complete denitrification generally occurred. Based on these observations, wood chip bioreactors may be successful at removing significant quantities of NO(3)-N, and reducing NO(3)-N concentration from water moving to subsurface drainage at flow rates observed in central Iowa subsoil. PMID- 19549944 TI - Effects of highway construction on stream water quality and macroinvertebrate condition in a mid-atlantic highlands watershed, USA. AB - Refining best management practices (BMPs) for future highway construction depends on a comprehensive understanding of environmental impacts from current construction methods. Based on a before-after-control impact (BACI) experimental design, long-term stream monitoring (1997-2006) was conducted at upstream (as control, n = 3) and downstream (as impact, n = 6) sites in the Lost River watershed of the Mid-Atlantic Highlands region, West Virginia. Monitoring data were analyzed to assess impacts of during and after highway construction on 15 water quality parameters and macroinvertebrate condition using the West Virginia stream condition index (WVSCI). Principal components analysis (PCA) identified regional primary water quality variances, and paired t tests and time series analysis detected seven highway construction-impacted water quality parameters which were mainly associated with the second principal component. In particular, impacts on turbidity, total suspended solids, and total iron during construction, impacts on chloride and sulfate during and after construction, and impacts on acidity and nitrate after construction were observed at the downstream sites. The construction had statistically significant impacts on macroinvertebrate index scores (i.e., WVSCI) after construction, but did not change the overall good biological condition. Implementing BMPs that address those construction-impacted water quality parameters can be an effective mitigation strategy for future highway construction in this highlands region. PMID- 19549945 TI - Long-term water quality trends after implementing best management practices in South Florida. AB - A mandatory best management practices (BMP) program was implemented in the Everglades Agricultural Area (EAA) farms basin-wide in 1995 as required by the Everglades Forever Act to reduce P loads in drainage water reaching the Everglades ecosystem. All farms in the EAA basin implement similar BMPs, and basin wide P load reductions have exceeded the 25% reduction required by law; however, differences remain in water quality between subbasins. Our objective was to determine long-term trends in P loads in discharge water in the EAA after implementing BMPs for 7 to10 yr and to explore reasons for differences in the performance of the subbasins. Two monitoring datasets were used, one from 10 research farms and the second from the EAA basin inflow and outflow locations. Mann-Kendall trend analysis was used to determine the degree of change in water quality trends. A decreasing trend in P loads was observed in general on sugarcane (Saccharum officinarum L.) farms, while mixed crop farms showed either decreasing or insignificant trends. The insignificant trends are probably related to management practices of mixed crop systems. Decreasing trends in P loads were observed in the outflow of the EAA basin, S5A, and S8 subbasins from 1992 to 2002. Inflow water from Lake Okeechobee had increasing P concentration from 1992 to 2006 with the highest trend in the east side of the lake. This analysis indicated there may be other factors impacting the success of BMPs in individual farms including cropping rotations and flooding of organic soils. Elevated P concentrations in Lake Okeechobee water used for irrigation may pose a future risk to degrade water quality on farms in the EAA, especially in the S5A subbasin. PMID- 19549946 TI - Removal of viruses and indicators by anaerobic membrane bioreactor treating animal waste. AB - Appropriate treatment of agricultural waste is necessary for the protection of public health in rural areas because land-applied animal manure may transmit zoonotic disease. In this study, we evaluated the potential of using a pilot anaerobic membrane bioreactor (AnMBR) to treat agricultural waste. The AnMBR system, following a conventional complete mix anaerobic digester (CMAD), achieved high removals of biological and chemical agents. The mean log(10) removals of Escherichia coli, enterococci, Clostridium perfringens, and coliphage by the AnMBR were 5.2, 6.1, 6.4, and 3.7, respectively, and for the CMAD were 1.5, 1.2, 0.1, and 0.5, respectively. Compared with other indicators, coliphage was observed most frequently and had the highest concentration in effluent samples. Bovine adenoviruses and bovine polymaviruses (BPyV) were monitored in this study using nested PCR methods. All of the CMAD influent and CMAD effluent samples were positive for both viruses, and three AnMBR effluent samples were BPyV positive. The mean removals of total Kjeldahl nitrogen, total phosphate, chemical oxygen demand, total solids, and volatile solids by the entire system were 31, 96, 92, 82, and 91%, respectively, but there was no removal of ammonium. When the AnMBR was operated independent of the CMAD, AnMBR achieved similar E. coli and enterococci removals as the combined CMAD/AnMBR system. The high quality of effluent produced by the pilot AnMBR system in this study demonstrated that such systems can be considered as alternatives for managing animal manure. PMID- 19549947 TI - Phytase supplemented poultry diets affect soluble phosphorus and nitrogen in manure and manure-amended soil. AB - Understanding P and N dynamics in manure-amended soil is essential for estimating the environmental impact of manure utilization in land applications. A laboratory incubation study was conducted to assess, (i) the effect of feeding a standard Australian commercial diet, and diets modified with phytase supplementation and reduced nonphytase phosphorus (NPP), on the concentrations of P and N (total and soluble) in the manure derived from layer hens (Gallus domesticus L.), and (ii) the change in water-soluble phoshorus (P(WSP)) and mineral N (NH(4)-N and NO(3) N) when used as a soil amendment, applied at rates equivalent to 200 kg ha(-1) (200N) and 400 kg ha(-1) (400N). Phytase supplementation increased %P(WSP) by 8 to 12% in the manures, regardless of the levels of NPP in the diets, and in the manure-amended soils by 27 to 30% at the 200N application rate, and up to 54% at the 400N rate. Phytase significantly (P < 0.05) reduced total nitrogen (TN) content (by 12-31%) of the manures but generally produced greater nitrate accumulation in the manure-amended soils. Net nitrification, which commenced 4 wk after incubation, was accompanied by a simultaneous decrease in soil pH (by one pH unit) and a concomitant decline in %P(WSP). The decline in %P(WSP) was primarily attributed to P retention by the soil as it became more acidic. This study suggests that phytase addition not only reduces manure total N content, and increases water-soluble P, but its effects on manure total phosphorus (TP) and 2 mol L(-1) KCl extractable mineral N is influenced by the NPP level in the diet. PMID- 19549948 TI - Effects of dried distillers' grains with solubles (wheat-based) in feedlot cattle diets on feces and manure composition. AB - The use of dried distillers' grains with solubles (DDGS) in feedlot cattle (Bos taurus) diets is increasing as the bio-ethanol industry expands. This study investigated how wheat (Triticum aestivum L.) DDGS-based diets impact feedlot cattle nutrient and volatile fatty acid (VFA) excretion. Feedlot heifers were fed DDGS at 0 (Control) 20, 40, 60% or 60% + Ca (1% limestone) of dietary dry matter. Feces and manure were sampled monthly over a 133-d finishing period. Total nitrogen (TN) (feces only), total phosphorus (TP), pH (manure only), and water soluble NH(4)(+) and P contents in feces and manure were higher with 40 and 60% DDGS diets than with the Control. Significant increases in isobutyric, valeric, and isovaleric VFAs (by far the most odorous in manure) were also observed in the feces with 40 and 60% DDGS diets, although there was no change in the total VFA content with diet. Wheat DDGS manure, with higher N and P contents, should be beneficial to crop production. However, it could potentially increase N and P loading on crop lands after application and contribute to greater NH(3) emission and malodor intensity while manure is in the feedlot pen. Estimated manure N loss while in feedlot pens also increased significantly with dietary DDGS levels. The small (nonsignificant) differences in total and soluble N and P in feces and manure between 20% DDGS and the Control (0% wheat DDGS) suggest that excess nutrient flow to the environment and malodors can be controlled by restricting wheat DDGS to a maximum of 20% in cattle diets. PMID- 19549949 TI - Transport of lincomycin to surface and ground water from manure-amended cropland. AB - Livestock manure containing antimicrobials becomes a possible source of these compounds to surface and ground waters when applied to cropland as a nutrient source. The potential for transport of the veterinary antimicrobial lincomycin to surface waters via surface runoff and to leach to ground water was assessed by monitoring manure-amended soil, simulated rainfall runoff, snowmelt runoff, and ground water over a 2-yr period in Saskatchewan, Canada, after fall application of liquid swine manure to cropland. Liquid chromatography tandem mass spectrometry was used to quantify lincomycin in all matrix extracts. Initial concentrations in soil (46.3-117 mug kg(-1)) were not significantly different (p > 0.05) for manure application rates ranging from 60,000 to 95,000 L ha(-1) and had decreased to nondetectable levels by mid-summer the following year. After fall manure application, lincomycin was present in all simulated rainfall runoff (0.07-2.7 mug L(-1)) and all snowmelt runoff (0.038-3.2 mug L(-1)) samples. Concentrations in snowmelt runoff were not significantly different from those in simulated rainfall runoff the previous fall. On average, lincomycin concentrations in ephemeral wetlands dissipated by 50% after 31 d. Concentrations of lincomycin in ground water were generally <0.005 mug L(-1). This study demonstrates that the management practice of using livestock manure from confined animal feeding operations as a plant nutrient source on cropland may result in antimicrobial transport to surface and ground waters. PMID- 19549950 TI - Fate of indicator microorganisms under nutrient management plan conditions. AB - Nutrient management plans (NMPs) for application of wastewater from concentrated animal feeding operations are designed to meet crop water and nutrient requirements, but implicitly assume that pathogenic microorganisms in the wastewater will be retained and die-off in the root zone. A NMP was implemented on a field plot to test this assumption by monitoring the fate of several fecal indicator microorganisms (Enterococcus, fecal coliforms, somatic coliphage, and total Escherichia coli). When well-water and wastewater were applied to meet measured evapotranspiration (ET), little advective transport of the indicator microorganisms occurred below the root zone and the remaining microorganisms rapidly died-off (within 1 mo). Additional experiments were conducted in the laboratory to better quantify microorganism transport and survival in the field soil. Batch survival experiments revealed much more rapid die-off rates for the bacterial indicator microorganisms in native than in sterilized soil, suggesting that biotic factors controlled survival. Saturated column experiments with packed field soil, demonstrated much greater transport potential for somatic coliphage than bacterial indicators (Enterococcus and total E. coli) and that the retention rates for the indicator microorganisms were not log-linear with depth. A worst case transport scenario of ponded infiltration on a large undistributed soil column from the field was also initiated and indicator microorganisms were not detected in the column outflow or in the soil at a depth of 65 cm. All of these observations support the hypothesis that a NMP at this site will protect groundwater supplies from microorganism contamination, especially when applied water and wastewater meet ET. PMID- 19549951 TI - Evaluation of second-generation multistage wastewater treatment system for the removal of malodors from liquid swine waste. AB - Wastewater quality and malodors in a second generation implementation of environmentally superior technology (EST) were monitored over three cycles of pig (Sus scrofa) production and 15 mo. The wastewater treatment system consisted of three modules: solids separation, biological N removal, and P recovery/wastewater disinfection. While approximately more than 90% of the wastewater suspended solids were removed in the first stage of treatment, little reduction in malodorous compounds occurred, indicating that malodors largely remained with the liquid waste stream. The greatest improvements in wastewater quality occurred in the N treatment module where ammonium was removed through nitrification/denitrification processes: there was more than 99% reduction in aromatic malodorous compounds (e.g., p-cresol, skatole) and almost 90% reduction in volatile fatty acids (e.g., propanoate and butanoate) in N module effluent as compared to raw flushed manure. The system performed consistently well in wastewater odor removal, even during the first cycle of livestock production when system performance was being optimized. These findings showed that the combination of the processes of solids removal and biological N treatment into a practical treatment system can be very effective in reducing malodors from livestock wastewater. PMID- 19549952 TI - Intake and digestibility of 'coastal' bermudagrass hay from treated swine waste using subsurface drip irrigation. AB - Waste handling systems for confined swine production in the upper South (approximately 32-37 degrees N and 79-93 degrees W) depend mainly on anaerobic lagoons and application of the waste effluent to cropland. The main objective of this study was to evaluate the quality of 'Coastal' bermudagrass [Cynodon dactylon (L.) Pers.] hay receiving effluent generated from a raw swine waste treatment system designed to reduce P and K concentrations and delivered by subsurface drip irrigation (SDI) compared with hay produced from commercial N fertilizer. Eight treatments, consisting of commercial N fertilizer or effluent, each irrigated at two irrigation rates (75 and 100% of estimated evapotranspiration) and two lateral spacings (0.6 and 1.2 m), were compared with a control treatment of commercial N fertilizer without irrigation. Three harvests were taken in each of 2 yr and five of the six evaluated using wether sheep (30 45 kg). Greatest dry matter intake (DMI) per unit body weight occurred for the control vs. all irrigated treatments (1.94 vs. 1.77 kg 100(-1) kg; P = 0.02; SEM = 0.11). Among irrigated treatments, DMI was greatest from commercial N vs. effluent (1.81 vs. 1.71 kg 100(-1) kg; P = 0.05; SEM = 0.11). Dry matter intake was similar for the 75% rate treatments and the non-irrigated treatment (mean, 1.87 kg 100(-1) kg) but was reduced for the 100% rate (1.94 vs. 1.72 kg 100(-1) kg; P = 0.03; SEM = 0.11). Hay from the 75% rate was more digestible than hay from the 100% rate (527 vs. 508 g kg(-1); P = 0.03; SEM = 21). The SDI system functioned well, and lateral spacing did not alter hay quality. Treated waste from a raw waste treatment system was readily delivered by SDI at the recommended rate to produce bermudagrass hay of adequate quality for ruminant production systems. PMID- 19549953 TI - Land use and riparian effects on prairie wetland sediment properties and herbicide sorption coefficients. AB - Sorption of commonly used herbicides by wetland sediment can provide important information for herbicide fate modeling. The influence of sediment properties on herbicide sorption as a result of different land uses in the wetland catchment is unclear. We examined the effects of land use on the physiochemical properties of wetland sediments and the associations between these sediment properties and herbicide sorption characteristics. Bottom sediments were sampled in 0- to 5- and 5- to 10-cm sections from 17 wetlands under five different land use classes: (i) ephemeral wetlands with no riparian vegetation in a cultivated catchment (ECNR), (ii) ephemeral wetlands with riparian vegetation in a cultivated catchment (ECR), (iii) ephemeral wetlands in a grassland catchment established 4 yr ago (E4G), (iv) ephemeral wetlands in a brome grass catchment established 20 yr ago (E20G), and (v) semi-permanent (SP) wetlands in a multiple-land-use catchment. Sediments were analyzed for total organic carbon (TOC), total inorganic carbon (TIC), pH, electrical conductivity, exchangeable cations (EXCAT), total cation exchangeable capacity (CEC), and percent clay (%clay). Sediment herbicide sorption partition coefficient (Kd) was measured for trifluralin, atrazine, 2,4-D, and glyphosate. The sorption of the herbicides in the sediment increased in the order of 2,4-D < atrazine < glyphosate < trifluralin. The sorption of 2,4-D, atrazine, and trifluralin was positively correlated to TOC, EXCAT, and CEC but negatively correlated to %clay. Glyphosate sorption was negatively correlated to pH, TIC, EXCAT, and %clay. Overall, wetland sediments that were recently cultivated (ECNR and E4G) had lower TOC, TIC, EC, EXCAT, CEC, and Kd values (2,4-D, trifluralin, and atrazine) than sediments that had not been recently cultivated (ECR, E20G, and SP). The ECR wetland sediments had the largest Kd for all four herbicides, suggesting that land use and riparian vegetation have a significant impact on herbicide sorption. PMID- 19549954 TI - Dissolved oxygen measurements in aquatic environments: the effects of changing temperature and pressure on three sensor technologies. AB - Dissolved oxygen (DO) is probably the most important parameter related to water quality and biological habitat in aquatic environments. In situ DO sensors are some of the most valuable tools used by scientists and engineers for the evaluation of water quality in aquatic ecosystems. Presently, we cannot accurately measure DO concentrations under variable temperature and pressure conditions. Pressure and temperature influence polarographic and optical type DO sensors compared to the standard Winkler titration method. This study combines laboratory and field experiments to compare and quantify the accuracy and performance of commercially available macro and micro Clark-type oxygen sensors as well as optical sensing technology to the Winkler method under changing pressure and temperature conditions. Field measurements at various lake depths revealed sensor response time up to 11 min due to changes in water temperature, pressure, and DO concentration. Investigators should account for transient response in DO sensors before measurements are collected at a given location. We have developed an effective model to predict the transient response time for Clark-type oxygen sensors. The proposed procedure increases the accuracy of DO data collected in situ for profiling applications. PMID- 19549955 TI - Comments on "Diversity in Cell Properties and Transport Behavior among 12 Different Environmental Escherichia coli Isolates," by C.H. Bolster, B.Z. Hazendaroglu, and S.L. Walker in the Journal of Environmental Quality 2009 38:465 472. PMID- 19549956 TI - Invitation or summons? UK debate surrounds messages about mammography. PMID- 19549957 TI - Pediatric brain tumor survivors, physicians, and researchers face long-term challenges. PMID- 19549958 TI - Does homeostatic pressure explain tumor growth? PMID- 19549959 TI - StatBite: Trends in U.S. childhood cancer survival (1975-2004). PMID- 19549961 TI - The cover. A hedgehog. PMID- 19549960 TI - Testing for carcinogens: shift from animals to automation gathers steam--slowly. PMID- 19549962 TI - A piece of my mind. "Minutes from fishing and the opera". PMID- 19549963 TI - Urology group: prostate screening should be offered beginning at age 40. PMID- 19549964 TI - States explore shared decision making. PMID- 19549965 TI - FDA tells drug and device makers to give balanced picture of risks in ads, labels. PMID- 19549966 TI - Economic conditions and obesity. PMID- 19549967 TI - Economic conditions and obesity. PMID- 19549968 TI - Evaluating care coordination among Medicare beneficiaries. PMID- 19549969 TI - Economic conditions and obesity. PMID- 19549970 TI - Clinical practice guideline development and antitrust law. PMID- 19549971 TI - Association of nipple piercing with abnormal milk production and breastfeeding. PMID- 19549972 TI - Body mass index and risk, age of onset, and survival in patients with pancreatic cancer. AB - CONTEXT: Obesity has been implicated as a risk factor for pancreatic cancer. OBJECTIVE: To demonstrate the association of excess body weight across an age cohort and the risk, age of onset, and overall survival of patients with pancreatic cancer. DESIGN, SETTING, AND PARTICIPANTS: A case-control study of 841 patients with pancreatic adenocarcinoma and 754 healthy individuals frequency matched by age, race, and sex. The study was conducted at a university cancer center in the United States from 2004 to 2008. Height and body weight histories were collected by personal interview starting at ages 14 to 19 years and over 10 year intervals progressing to the year prior to recruitment in the study. MAIN OUTCOME MEASURES: The associations between patients' body mass index (BMI) and risk of pancreatic cancer, age at onset, and overall survival were examined by unconditional logistic regression, linear regression, and Cox proportional hazard regression models, respectively. RESULTS: Individuals who were overweight (a BMI of 25-29.9) from the ages of 14 to 39 years (highest odds ratio [OR], 1.67; 95% confidence interval [CI], 1.20-2.34) or obese (a BMI > or = 30) from the ages of 20 to 49 years (highest OR, 2.58; 95% CI, 1.70-3.90) had an associated increased risk of pancreatic cancer, independent of diabetes status. The association was stronger in men (adjusted OR, 1.80; 95% CI, 1.45-2.23) by mean BMI from the ages of 14 to 59 years than in women (adjusted OR, 1.32; 95% CI, 1.02-1.70) and in ever smokers (adjusted OR, 1.75; 95% CI, 1.37-2.22) than in never smokers (adjusted OR, 1.46; 95% CI, 1.16-1.84). The population-attributable risk percentage of pancreatic cancer based on the mean BMI from the ages of 14 to 59 years was 10.3% for never smokers and 21.3% for ever smokers. Individuals who were overweight or obese from the ages of 20 to 49 years had an earlier onset of pancreatic cancer by 2 to 6 years (median age of onset was 64 years for patients with normal weight, 61 years for overweight patients [P = .02], and 59 years for obese patients [P < .001]). Compared with those with normal body weight and after adjusting for all clinical factors, individuals who were overweight or obese from the ages of 30 to 79 years or in the year prior to recruitment had reduced overall survival of pancreatic cancer regardless of disease stage and tumor resection status (overweight patients: hazard ratio, 1.26 [95% CI, 0.94-1.69], P = .04; obese patients: hazard ratio, 1.86 [95% CI, 1.35-2.56], P < .001). CONCLUSIONS: Overweight or obesity during early adulthood was associated with a greater risk of pancreatic cancer and a younger age of disease onset. Obesity at an older age was associated with a lower overall survival in patients with pancreatic cancer. PMID- 19549973 TI - Migraine headache in middle age and late-life brain infarcts. AB - CONTEXT: Migraine is considered to be an episodic condition with no long-term consequences. However, recent studies suggest that migraine attacks may be associated with pathologic changes in the brain, particularly in the cerebellum. OBJECTIVE: To determine whether individuals not reporting headache compared with individuals reporting migraine symptoms, particularly aura, in midlife are at increased risk of late-life infarct-like lesions found on magnetic resonance imaging (MRI) without consideration of clinical symptoms. DESIGN, SETTING, AND PARTICIPANTS: A population-based study of men and women in Reykjavik, Iceland (cohort born 1907-1935; n = 4689; 57% women) were followed up since 1967, examined, and interviewed about migraine symptoms in midlife (mean age, 51 years; range, 33-65 years). Between 2002 and 2006, more than 26 years later, brain MRIs were performed. Participants reporting headaches once or more per month were asked about migraine symptoms including nausea, unilateral location, photophobia, visual disturbance, and numbness. These individuals with headache were classified as having migraine without aura, migraine with aura, or nonmigraine headache. A comprehensive cardiovascular risk assessment was performed at both examinations. MAIN OUTCOME MEASURE: Presence of infarct-like lesions (total) and specifically located in the cortical, subcortical, and cerebellar regions. RESULTS: Infarct like lesions were present in 39.3% of men and 24.6% of women. After adjusting for age, sex, and follow-up time, compared with those not reporting headaches once or more per month (n = 3243), those with midlife migraine with aura (n = 361) had an increased risk of late-life infarct-like lesions (adjusted odds ratio [OR], 1.4; 95% confidence interval [CI], 1.1-1.8) that specifically reflected an association with cerebellar lesions in women (prevalence of infarcts 23.0% for women with migraine with aura vs 14.5% for women not reporting headaches; adjusted OR, 1.9; 95% CI, 1.4-2.6 vs a 19.3% prevalence of infarcts for men with migraine with aura vs 21.3% for men not reporting headaches; adjusted OR, 1.0; 95% CI, 0.6-1.8; P<.04 for interaction by sex). Migraine without aura and nonmigraine headache were not associated with an increased risk. CONCLUSIONS: Migraine with aura in midlife was associated with late-life prevalence of cerebellar infarct-like lesions on MRI. This association was statistically significant only for women. This is consistent with the hypothesis that migraine with aura in midlife is associated with late-life vascular disease in the cerebellum and in women. PMID- 19549974 TI - Long-term outcomes in individuals with prolonged PR interval or first-degree atrioventricular block. AB - CONTEXT: Prolongation of the electrocardiographic PR interval, known as first degree atrioventricular block when the PR interval exceeds 200 milliseconds, is frequently encountered in clinical practice. OBJECTIVE: To determine the clinical significance of PR prolongation in ambulatory individuals. DESIGN, SETTING, AND PARTICIPANTS: Prospective, community-based cohort including 7575 individuals from the Framingham Heart Study (mean age, 47 years; 54% women) who underwent routine 12-lead electrocardiography. The study cohort underwent prospective follow-up through 2007 from baseline examinations in 1968-1974. Multivariable-adjusted Cox proportional hazards models were used to examine the associations of PR interval with the incidence of arrhythmic events and death. MAIN OUTCOME MEASURES: Incident atrial fibrillation (AF), pacemaker implantation, and all-cause mortality. RESULTS: During follow-up, 481 participants developed AF, 124 required pacemaker implantation, and 1739 died. At the baseline examination, 124 individuals had PR intervals longer than 200 milliseconds. For those with PR intervals longer than 200 milliseconds compared with those with PR intervals of 200 milliseconds or shorter, incidence rates per 10 000 person-years were 140 (95% confidence interval [CI], 95-208) vs 36 (95% CI, 32-39) for AF, 59 (95% CI, 40-87) vs 6 (95% CI, 5-7) for pacemaker implantation, and 334 (95% CI, 260-428) vs 129 (95% CI, 123-135) for all-cause mortality. Corresponding absolute risk increases were 1.04% (AF), 0.53% (pacemaker implantation), and 2.05% (all-cause mortality) per year. In multivariable analyses, each 20-millisecond increment in PR was associated with an adjusted hazard ratio (HR) of 1.11 (95% CI, 1.02-1.22; P = .02) for AF, 1.22 (95% CI, 1.14-1.30; P < .001) for pacemaker implantation, and 1.08 (95% CI, 1.02-1.13; P = .005) for all-cause mortality. Individuals with first-degree atrioventricular block had a 2-fold adjusted risk of AF (HR, 2.06; 95% CI, 1.36-3.12; P < .001), 3-fold adjusted risk of pacemaker implantation (HR, 2.89; 95% CI, 1.83-4.57; P < .001), and 1.4-fold adjusted risk of all-cause mortality (HR, 1.44, 95% CI, 1.09-1.91; P = .01). CONCLUSION: Prolongation of the PR interval is associated with increased risks of AF, pacemaker implantation, and all-cause mortality. PMID- 19549975 TI - Evolving health effects of Pneumocystis: one hundred years of progress in diagnosis and treatment. AB - 2009 marks the 100th anniversary of the first description of Pneumocystis, an organism that was ignored for much of its first 50 years but that has subsequently been recognized as an important pathogen of immunocompromised patients, especially patients infected with human immunodeficiency virus (HIV). We present a patient with chronic lymphocytic leukemia who died from Pneumocystis pneumonia (PCP) despite appropriate anti-Pneumocystis therapy. Although substantial advances in diagnosis, treatment, and prevention of PCP have decreased its frequency and improved prognosis, PCP continues to be seen in both HIV-infected patients and patients receiving immunosuppressive medications. Pneumocystis species comprise a family of fungi, each of which appears to be able to infect only 1 host species. Pneumocystis has a worldwide distribution. Immunocompetent hosts clear infection without obvious clinical consequences. Pneumocystis has been identified in patients with other diseases such as chronic obstructive pulmonary disease, although its clinical impact is uncertain. Immunocompromised patients develop disease as a consequence of reinfection and possibly reactivation of latent infection. In patients with HIV infection, the CD4 count is predictive of the risk for developing PCP, but such reliable markers are not available for other immunocompromised populations. In the majority of patients with PCP, multiple Pneumocystis strains can be identified using recently developed typing techniques. Because Pneumocystis cannot be cultured, diagnosis relies on detection of the organism by colorimetric or immunofluorescent stains or by polymerase chain reaction. Trimethoprim-sulfamethoxazole is the preferred drug regimen for both treatment and prevention of PCP, although a number of alternatives are also available. Corticosteroids are an important adjunct for hypoxemic patients. PMID- 19549976 TI - Evolutionary speculation about tuberculosis and the metabolic and inflammatory processes of obesity. PMID- 19549977 TI - A health care cooperative extension service: transforming primary care and community health. PMID- 19549978 TI - Overweight, obesity, and pancreatic cancer: beyond risk alone. PMID- 19549979 TI - Migraine and cerebral infarct-like lesions on MRI: an observation, not a disease. PMID- 19549980 TI - JAMA patient page. Migraine headache. PMID- 19549981 TI - Gait speed under varied challenges and cognitive decline in older persons: a prospective study. AB - OBJECTIVE: to examine whether usual gait speed, fast gait speed or speed while walking with a cognitive or neuromuscular challenge predicts evolving cognitive decline over 3 years. DESIGN: prospective study. SETTING: population-based sample of community-dwelling older persons. PARTICIPANTS: 660 older participants (age > or = 65 years). MEASUREMENTS: usual gait speed, fastest gait speed, gait speed during 'walking-while-talking', depression, comorbidities, education, smoking and demographics were assessed at baseline. Cognition was evaluated at baseline and follow-up. A decline in MMSE score by > or = 3 points was considered as significant cognitive decline (SCD). RESULTS: adjusting for confounders, only fast speed was associated with cognitive performance at 3-year follow-up. One hundred thirty-five participants had SCD over 3 years. Participants in the lowest quartile of usual speed or walking-while-talking speed were more likely to develop SCD. Conversely, participants in the third and fourth quartiles of fast speed were more likely to develop SCD. J-test showed that the model including fast speed quartiles as a regressor was significantly more predictive of SCD than the models with usual speed or walking-while-talking speed quartiles. CONCLUSION: measuring fast gait speed in older persons may assist in identifying those at high risk of cognitive decline. PMID- 19549983 TI - Activation of T cells: releasing the brakes by proteolytic elimination of Cbl-b. AB - Activation of T lymphocytes relies on the simultaneous delivery of signals from the T cell receptor and co-receptors such as CD28. The absence of co-receptor signaling leads to a state of unresponsiveness called anergy, which prevents T cells from reacting against self antigens. The biochemical program that ensures the induction of anergy uses several mechanisms, including the synthesis of ubiquitin E3 ligases such as Cbl-b (Casitas B-lineage lymphoma). Because these E3 ligases function as gatekeepers to prevent the undesired activation of T cells, full and productive induction of the T cell response requires the restriction of these negative regulators by mechanisms that we are beginning to understand. PMID- 19549984 TI - The kinesin protein Kif7 is a critical regulator of Gli transcription factors in mammalian hedgehog signaling. AB - From insects to humans, the Hedgehog (Hh) signaling pathway has conserved roles in embryonic development and tissue homeostasis. However, it has been suggested that the lack of mammalian equivalents of Costal2 (Cos2) contributes to a divergence between the mechanism of Drosophila and mammalian Hh signal transduction. Here, we challenge this view by showing that the kinesin protein Kif7 is a critical regulator of Hh signaling in mice. Similar to Cos2, Kif7 physically interacted with Gli transcription factors and controlled their proteolysis and stability, and acted both positively and negatively in Hh signaling. Thus, Kif7 is a missing component of the mammalian Hh signaling machinery, implying a greater commonality between the Drosophila and mammalian system than the prevailing view suggests. PMID- 19549985 TI - PKC-theta modulates the strength of T cell responses by targeting Cbl-b for ubiquitination and degradation. AB - The E3 ubiquitin ligase Casitas B-lineage lymphoma (Cbl-b) is central to antigen induced immune tolerance and regulates the CD28 dependence of T cell activation. Cbl-b undergoes ubiquitination and proteasomal degradation after adequate costimulation of T cells; however, the mechanism involved is unknown. Here, we identified protein kinase C-theta (PKC-theta) as the critical intermediary for the inactivation of Cbl-b in response to costimulation of T cells through CD28. PKC-theta associated with Cbl-b on stimulation of the T cell receptor. After costimulation of T cells through CD28, Cbl-b was ubiquitinated and degraded through a mechanism that depended on the kinase activity of PKC-theta. Consistent with this mechanism, the impaired responses of PKCtheta-deficient T cells were at least partially restored by the concomitant genetic loss of cblb. Thus, our data establish a nonredundant antagonism between PKC-theta and Cbl-b that regulates T cell activation responses. PMID- 19549987 TI - Risk of arterial cardiovascular events in patients after pulmonary embolism. AB - Studies have reported inconsistent evidence for an association between venous thrombosis and arterial cardiovascular events. We further studied the association between both diseases by comparing the occurrence of cardiovascular events in patients diagnosed with acute pulmonary embolism (PE) contrasted to patients with comparable baseline risk characteristics (patients in whom PE was clinically suspected but ruled out). Included were 259 patients with provoked PE, 95 patients with unprovoked PE, and 334 control patients without PE. Patients diagnosed with PE were treated with vitamin K antagonists for 6 months. Median follow-up was 4.2 years. Sixty-three arterial cardiovascular events were registered (incidence, 5.1/100 patient-years). Adjusted hazard ratio was not different between patients with all-cause PE and control patients (1.39, 95% confidence interval [CI], 0.83-2.3) but increased for patients with unprovoked PE versus both patients with provoked PE and control patients without PE (2.18; 95% CI, 1.1-4.5; and 2.62; 95% CI, 1.4-4.9, respectively). This effect was confirmed after redefining the study start date to the moment the vitamin K antagonists were discontinued. Our study underlines the association between unprovoked venous thrombosis and arterial cardiovascular events; however, risk differences between patients with provoked PE and patients in whom PE was clinically suspected but ruled out could not be demonstrated. PMID- 19549986 TI - Lineages of human T-cell clones, including T helper 17/T helper 1 cells, isolated at different stages of anti-factor VIII immune responses. AB - The development of neutralizing antibodies (inhibitors) after factor VIII (FVIII) infusions is a serious complication that affects approximately one-quarter of hemophilia A patients who have access to replacement therapy. To investigate the differentiation of naive T cells into FVIII-specific helper T cells that promote B-cell activation and antibody secretion, HLA-DRA-DRB1*0101-restricted T-cell clones that respond to a specific epitope in FVIII were isolated from a mild hemophilia A subject (the proband) 19 weeks and 21 months after his development of a high-titer inhibitor. Clones responding to the same epitope were also isolated from his multiply infused brother, who has not developed a clinically significant inhibitor. The 19-week proband clones were T helper (T(H))17/T(H)1- or T(H)1/T(H)2-polarized, whereas all 8 clones isolated 21 months postinhibitor development were T(H)2-polarized cells. In contrast, all 6 clones from the brother who did not develop an inhibitor were T(H)1-polarized, indicating that tolerance to FVIII can be maintained even with circulating T(H)1-polarized cells that respond vigorously to in vitro FVIII stimulation. This is the first evidence that T(H)17/T(H)1-polarized cells play a role in hemophilic immune responses to FVIII. Furthermore, this is the first report of successful isolation and expansion of antigen-specific human T(H)17/T(H)1 clones using standard culture conditions. PMID- 19549988 TI - Identification of patients with poorer survival in primary myelofibrosis based on the burden of JAK2V617F mutated allele. AB - A total of 186 patients with primary myelofibrosis (PMF) were genotyped for JAK2V617F at diagnosis aimed at analyzing the correlation of mutational status and mutated allele burden with outcome variables, including time to anemia, leukocytosis, leukopenia, thrombocytopenia, massive splenomegaly, leukemia, and with overall survival. A total of 127 JAK2V617F-mutated patients (68% of whole series) were divided in quartiles of V617F allele burden. After a median follow up of 17.2 months, 23 patients died, 15 because of leukemia. A JAK2V617F mutated status did not impact on the rate of leukemia transformation or overall survival. Patients in the lower quartile had shorter time to anemia and leukopenia and did not progress to large splenomegaly. Furthermore, survival was significantly reduced in the lower quartile compared with upper quartiles and JAK2 wild-type patients. In multivariate analysis, factors associated with reduced survival were age, a blast count more than 1%, and a JAK2V617F burden within first quartile. Causes of death in the lower quartile were represented mainly by systemic infections. We conclude that a low JAK2V617F allele burden at diagnosis is preferentially associated with a myelodepletive rather than myeloproliferative phenotype and represents an independent factor associated with shortened survival in patients with PMF. PMID- 19549989 TI - Absence of collagen-induced platelet activation caused by compound heterozygous GPVI mutations. AB - The glycoprotein VI (GPVI)/FcRgamma complex is a key receptor for platelet activation by collagen. We describe, for the first time, 2 genetic abnormalities in one patient. This 10-year-old girl presented ecchymoses since infancy, a prolonged bleeding time despite a normal platelet count and no antiplatelet antibodies. Collagen-induced platelet activation was null, whereas GPVI quantification by flow cytometry evidenced an incomplete deficiency. Immunoblotting showed an abnormal migration of residual GPVI, and no FcRgamma defect. GPVI DNA sequencing revealed (1) an R38C mutation in exon 3 of one allele and (2) an insertion of 5 nucleotides in exon 4 of the other allele, leading to a premature nonsense codon and absence of the corresponding mRNA. Introduction of the R38C mutation into recombinant GPVI-Fc resulted in abnormal protein migration and a loss of collagen binding. Thus, this composite genetic GPVI deficiency and dysfunction cause absence of platelet responses to collagen and a mild bleeding phenotype. PMID- 19549990 TI - The supernatant of apoptotic cells causes transcriptional activation of hypoxia inducible factor-1alpha in macrophages via sphingosine-1-phosphate and transforming growth factor-beta. AB - Macrophages infiltrating solid tumors exhibit a tumor-supporting phenotype and are critical for tumor development. Little is known which tumor-derived signal provokes this phenotype shift and how these signals are interpreted in macrophages to support tumor growth. We used the supernatant of apoptotic cells and noticed transcriptional, nuclear factor of activated T cells-dependent up regulation of hypoxia-inducible factor (HIF)-1alpha mRNA, subsequent protein expression, and HIF-1 activity. Blocking calcineurin with cyclosporine A attenuated nuclear factor of activated T cells binding during electrophoretic mobility shift assay analysis and circumvented the HIF-1alpha mRNA increase. Knockdown experiments, receptor analysis, and antibody neutralization pointed to sphingosine-1-phosphate and transforming growth factor-beta as the initiators of the HIF-1 response. The use of macrophages from conditional HIF-1alpha knockout mice revealed that macrophages, under the impact of apoptotic cell supernatants, use HIF-1 to produce factors that induce CD31 expression in murine embryonic stem cells. Our study supports the notion that soluble factors produced from apoptotic tumor cells activate the HIF-1 system under normoxia in macrophages to enhance their tumor-promoting capacity by, for example, releasing vascular endothelial growth factor. This shows the importance of HIF-1-elicited responses in regulatory macrophages under normoxia. PMID- 19549991 TI - FGF23 is mainly synthesized by osteocytes in the regularly distributed osteocytic lacunar canalicular system established after physiological bone remodeling. AB - This study aimed to evaluate whether the immunolocalization of fibroblast growth factor (FGF) 23 and dentin matrix protein 1 (DMP1) is associated with the spatial regularity of the osteocyte lacunar canalicular system(s) (OLCS). Femora of 12 weeks-old male ICR mice were fixed with 4% paraformaldehyde, decalcified with a 10% EDTA solution and then embedded in paraffin. We have devised a triple staining procedure that combines silver impregnation, alkaline phosphatase (ALPase) immunohistochemistry and tartrate-resistant acid phosphatase (TRAPase) enzyme histochemistry on a single paraffin section. This procedure permitted the visualization of ALPase-positive plump osteoblasts and several TRAPase-positive osteoclasts on those bone matrices featuring irregularly arranged OLCS, and of ALPase-positive bone lining cells on the bone matrix displaying the well-arranged OLCS. As observations proceeded from the metaphysis toward the diaphysis, the endosteal cortical bone displayed narrower bands of calcein labeling, accompanied by increased regularity of the OLCS. This implies that the speed of bone deposition during bone remodeling would affect the regularity of the OLCS. While DMP1 was evenly localized in all regions of the cortical bones, FGF23 was more abundantly localized in osteocytes of cortical bones with regularly arranged OLCS. In cortical bones, the endosteal area featuring regular OLCS exhibited more intense FGF23 immunoreaction when compared to the periosteal region, which tended to display irregular OLCS. In summary, FGF23 appears to be synthesized principally by osteocytes in the regularly distributed OLCS that have been established after bone remodeling. PMID- 19549992 TI - When older African American women are affected by violence in the home: a qualitative investigation of risk and protective factors. AB - A growing body of work has begun to recognize that the problem of older women affected by family violence (FV) deserves special consideration. Because risk and protective factors for FV can vary by social and cultural context, it is imperative that more focused attention be paid to vulnerable populations such as older African American women. This article reports on the results of a qualitative inquiry into individual, dyadic, and community-level variables that influence the risk of FV among older African American women receiving primary care at an inner-city hospital. An explanatory model that integrates these variables is presented. PMID- 19549993 TI - Group health cooperative's transformation toward patient-centered access. AB - The Institute of Medicine suggests redesigning health care to ensure safe, effective, timely, efficient, equitable, and patient-centered care. The concept of patient-centered access supports these goals. Group Health, a mixed-model health care system, attempted to improve patients' access to care through the following changes: (a) offering a patient Web site with patient access to patient physician secure e-mail, electronic medical records, and health promotion information; (b) offering advanced access to primary physicians; (c) redesigning primary care services to enhance care efficiency; (d) offering direct access to physician specialists; and (e) aligning primary physician compensation through incentives for patient satisfaction, productivity, and secure messaging with patients. In the 2 years following the redesign, patients reported higher satisfaction with certain aspects of access to care, providers reported improvements in the quality of service given to patients, and enrollment in Group Health stayed aligned with statewide trends in health care coverage. PMID- 19549994 TI - Monitoring bone mineral density during antiresorptive treatment for osteoporosis. PMID- 19549995 TI - Variation in antibiotic prescribing and its impact on recovery in patients with acute cough in primary care: prospective study in 13 countries. AB - OBJECTIVE: To describe variation in antibiotic prescribing for acute cough in contrasting European settings and the impact on recovery. DESIGN: Cross sectional observational study with clinicians from 14 primary care research networks in 13 European countries who recorded symptoms on presentation and management. Patients followed up for 28 days with patient diaries. SETTING: Primary care. PARTICIPANTS: Adults with a new or worsening cough or clinical presentation suggestive of lower respiratory tract infection. MAIN OUTCOME MEASURES: Prescribing of antibiotics by clinicians and total symptom severity scores over time. RESULTS: 3402 patients were recruited (clinicians completed a case report form for 99% (3368) of participants and 80% (2714) returned a symptom diary). Mean symptom severity scores at presentation ranged from 19 (scale range 0 to 100) in networks based in Spain and Italy to 38 in the network based in Sweden. Antibiotic prescribing by networks ranged from 20% to nearly 90% (53% overall), with wide variation in classes of antibiotics prescribed. Amoxicillin was overall the most common antibiotic prescribed, but this ranged from 3% of antibiotics prescribed in the Norwegian network to 83% in the English network. While fluoroquinolones were not prescribed at all in three networks, they were prescribed for 18% in the Milan network. After adjustment for clinical presentation and demographics, considerable differences remained in antibiotic prescribing, ranging from Norway (odds ratio 0.18, 95% confidence interval 0.11 to 0.30) to Slovakia (11.2, 6.20 to 20.27) compared with the overall mean (proportion prescribed: 0.53). The rate of recovery was similar for patients who were and were not prescribed antibiotics (coefficient -0.01, P<0.01) once clinical presentation was taken into account. CONCLUSIONS: Variation in clinical presentation does not explain the considerable variation in antibiotic prescribing for acute cough in Europe. Variation in antibiotic prescribing is not associated with clinically important differences in recovery. TRIAL REGISTRATION: Clinicaltrials.gov NCT00353951. PMID- 19549998 TI - Quantitative myotonia assessment using force relaxation curve modelling. AB - The lack of a robust quantitative measure of myotonia has been underlined in previous studies. Recent publications have proposed methods to quantify myotonia based on the measurement of force relaxation times during maximal contractions. However, they present several drawbacks mainly due to unstable force, odd peaks or digital noise. A possible solution to this issue consists in fitting the force curve with a convenient regression model. The aim of this study was, therefore, to provide a regression model in order to fit the force relaxation time curve automatically and to provide a robust index for quantitative assessment of myotonia in clinical settings. Force curves were fitted by an asymmetric sigmoidal function. The inverse function was then used to compute various absolute and relative relaxation times automatically. These variables were calculated for 16 controls and 16 patients with myotonic dystrophy type 1 (DM1). All variables were significantly increased in DM1 patients compared to controls. For instance, the relaxation time between 40 and 60% of the initial contraction level was 18.2 (SD: 3.3) ms in controls and 40.1 (SD: 17.7) ms in DM1 patients. All relaxation variables were highly discriminant. Force curve modelling provides an objective and effective quantification of myotonia. PMID- 19549996 TI - Value of routine monitoring of bone mineral density after starting bisphosphonate treatment: secondary analysis of trial data. AB - OBJECTIVE: To assess the value of monitoring response to bisphosphonate treatment by means of measuring bone mineral density. DESIGN: Secondary analysis of trial data using mixed models. Data source The Fracture Intervention Trial, a randomised controlled trial that compared the effects of alendronate and placebo in 6459 postmenopausal women with low bone mineral density recruited between May 1992 and May 1993. Bone density measurements of hip and spine were obtained at baseline and at one, two, and three years after randomisation. MAIN OUTCOME MEASURES: Between-person (treatment related) variation and within-person (measurement related) variation in hip and spine bone mineral density. RESULTS: The mean effect of three years' treatment with alendronate was to increase hip bone mineral density by 0.030 g/cm(2). There was some between-person variation in the effects of alendronate, but this was small in size compared with within person variation. Alendronate treatment is estimated to result in increases in hip bone density >or=0.019 g/cm(2) in 97.5% of patients. CONCLUSIONS: Monitoring bone mineral density in postmenopausal women in the first three years after starting treatment with a potent bisphosphonate is unnecessary and may be misleading. Routine monitoring should be avoided in this early period after bisphosphonate treatment is commenced. PMID- 19549999 TI - Conformational energies and entropies of peptides, and the peptide-protein binding problem. AB - A novel statistical thermodynamic approach is applied to free-peptide segments in order to classify them according to their conformational energies, entropies and heat capacities. Our approach employs the rotational isomeric state (RIS) model in which the states are described by the Ramachandran map of backbone torsion angles. The statistical weight matrices for the pairwise-dependent states are derived from the torsion angle probabilities of the consecutive dipeptides in a coil library. The partition function is determined for a given sequence via RIS multiplication of the pre-determined matrices. The conformational partition function, Helmholtz free energy, energy, entropy and heat capacity are obtained. The model is applied to randomly produced peptides and also to known peptide inhibitors to analyze their thermodynamic properties. Peptides with low energy, low entropy and low-heat capacity are determined to be essential for a peptide to be a good candidate inhibitor. Free energy changes in peptide binding are also discussed. PMID- 19549997 TI - Anatomy of health effects of Mediterranean diet: Greek EPIC prospective cohort study. AB - OBJECTIVE: To investigate the relative importance of the individual components of the Mediterranean diet in generating the inverse association of increased adherence to this diet and overall mortality. DESIGN: Prospective cohort study. SETTING: Greek segment of the European Prospective Investigation into Cancer and nutrition (EPIC). PARTICIPANTS: 23 349 men and women, not previously diagnosed with cancer, coronary heart disease, or diabetes, with documented survival status until June 2008 and complete information on nutritional variables and important covariates at enrolment. MAIN OUTCOME MEASURE: All cause mortality. RESULTS: After a mean follow-up of 8.5 years, 652 deaths from any cause had occurred among 12 694 participants with Mediterranean diet scores 0-4 and 423 among 10 655 participants with scores of 5 or more. Controlling for potential confounders, higher adherence to a Mediterranean diet was associated with a statistically significant reduction in total mortality (adjusted mortality ratio per two unit increase in score 0.864, 95% confidence interval 0.802 to 0.932). The contributions of the individual components of the Mediterranean diet to this association were moderate ethanol consumption 23.5%, low consumption of meat and meat products 16.6%, high vegetable consumption 16.2%, high fruit and nut consumption 11.2%, high monounsaturated to saturated lipid ratio 10.6%, and high legume consumption 9.7%. The contributions of high cereal consumption and low dairy consumption were minimal, whereas high fish and seafood consumption was associated with a non-significant increase in mortality ratio. CONCLUSION: The dominant components of the Mediterranean diet score as a predictor of lower mortality are moderate consumption of ethanol, low consumption of meat and meat products, and high consumption of vegetables, fruits and nuts, olive oil, and legumes. Minimal contributions were found for cereals and dairy products, possibly because they are heterogeneous categories of foods with differential health effects, and for fish and seafood, the intake of which is low in this population. PMID- 19550000 TI - Real-time motion-adaptive-optimization (MAO) in TomoTherapy. AB - IMRT delivery follows a planned leaf sequence, which is optimized before treatment delivery. However, it is hard to model real-time variations, such as respiration, in the planning procedure. In this paper, we propose a negative feedback system of IMRT delivery that incorporates real-time optimization to account for intra-fraction motion. Specifically, we developed a feasible workflow of real-time motion-adaptive-optimization (MAO) for TomoTherapy delivery. TomoTherapy delivery is characterized by thousands of projections with a fast projection rate and ultra-fast binary leaf motion. The technique of MAO-guided delivery calculates (i) the motion-encoded dose that has been delivered up to any given projection during the delivery and (ii) the future dose that will be delivered based on the estimated motion probability and future fluence map. These two pieces of information are then used to optimize the leaf open time of the upcoming projection right before its delivery. It consists of several real-time procedures, including 'motion detection and prediction', 'delivered dose accumulation', 'future dose estimation' and 'projection optimization'. Real-time MAO requires that all procedures are executed in time less than the duration of a projection. We implemented and tested this technique using a TomoTherapy research system. The MAO calculation took about 100 ms per projection. We calculated and compared MAO-guided delivery with two other types of delivery, motion-without compensation delivery (MD) and static delivery (SD), using simulated 1D cases, real TomoTherapy plans and the motion traces from clinical lung and prostate patients. The results showed that the proposed technique effectively compensated for motion errors of all test cases. Dose distributions and DVHs of MAO-guided delivery approached those of SD, for regular and irregular respiration with a peak-to-peak amplitude of 3 cm, and for medium and large prostate motions. The results conceptually proved that the proposed method is applicable for real-time motion compensation in TomoTherapy delivery. Extension of the method to real-time adaptive radiation therapy (ART) that compensates for all kinds of delivery errors was proposed. Further validation and clinical implementation is underway. PMID- 19550001 TI - Further factors for the estimation of mean glandular dose using the United Kingdom, European and IAEA breast dosimetry protocols. AB - The United Kingdom, European and IAEA protocols for breast dosimetry in mammography make use of s-factors which allow for the use of different target/filter combinations. To supplement the existing protocols, a Monte Carlo computer program has been used to calculate s-factors for mammography using a tungsten target with silver filters of thicknesses 50-75 microm and for the same target filtered with 0.5 mm aluminium. The dosimetry protocols use slabs of polymethyl methacrylate (PMMA) of specified thicknesses to simulate the exposure of typical breasts. The equivalent thickness of PMMA has been calculated using a simplified approach for a wider range of x-ray spectra and for breast thicknesses of 2-11 cm. The results show that for the tungsten/silver target/filter combination, a single s-factor of 1.042 can be used with the protocols, but when the tungsten target is filtered with 0.5 mm of aluminium, it is necessary to select from a tabulation of s-factors against breast thickness. The equivalent thicknesses of PMMA for a given breast thickness show some dependence on beam quality and the values obtained differ from those presently used in the dosimetry protocols by an amount which depends upon breast thickness and half value layer (HVL). For the extreme case of an 11 cm breast and an HVL of 0.62 mm Al, the use of the protocol thickness would give rise to an error of 10%, but for breast thicknesses of 6 cm or less, the error is typically 2-3%. PMID- 19550002 TI - Breathing interplay effects during proton beam scanning: simulation and statistical analysis. AB - Treatment delivery with active beam scanning in proton radiation therapy introduces the problem of interplay effects when pencil beam motion occurs on a similar time scale as intra-fractional tumor motion. In situations where fractionation may not provide enough repetition to blur the effects of interplay, repeated delivery or 'repainting' of each field several times within a fraction has been suggested. The purpose of this work was to investigate the effectiveness of different repainting strategies in proton beam scanning. To assess the dosimetric impact of interplay effects, we performed a series of simulations considering the following parameters: tumor motion amplitude, breathing period, asymmetry in the motion trajectory for the target and time required to change the beam energy for the delivery system. Several repainting strategies were compared in terms of potential vulnerability to a dose delivery error. Breathing motion perpendicular to the beam direction (representing superior-inferior type tumor motion in patients) was considered and modeled as an asymmetric sine function with a peak-to-peak amplitude of between 10 and 30 mm. The results show that motion effects cause a narrowing of the high-dose profile and widening of the penumbra. The 90% isodose area was reduced significantly when considering a large motion amplitude of 3 cm. The broadening of the penumbra appears to depend only on the amplitude of tumor motion (assuming harmonic motion). The delivered dose exhibits a shift of 10-15% of the tumor amplitude (or 1-5 mm) in the caudal direction due to breathing asymmetry observed for both sin(4)(x) and sin(6)(x) motion. Of the five repainting techniques studied, so-called 'breath sampling' turned out to be most effective in reducing dose errors with a minimal increase in treatment time. In this method, each energy level is repainted at several evenly spaced times within one breathing period. To keep dose delivery errors below 5% while minimizing treatment time, it is recommended that breath sampling repainting be employed using 5-10 paintings per field for an assumed tumor volume of 8.5 x 8.5 x 10 cm(3). For smaller tumor volumes more repaintings will be required, while for larger volumes five repaintings should be sufficient to achieve the required dose accuracy. PMID- 19550003 TI - Comparison of surface doses from spot scanning and passively scattered proton therapy beams. AB - Proton therapy for the treatment of cancer is delivered using either passively scattered or scanning beams. Each technique delivers a different amount of dose to the skin, because of the specific feature of their delivery system. The amount of dose delivered to the skin can play an important role in choosing the delivery technique for a specific site. To assess the differences in skin doses, we measured the surface doses associated with these two techniques. For the purpose of this investigation, the surface doses in a phantom were measured for ten prostate treatment fields planned with passively scattered proton beams and ten patients planned with spot scanning proton beams. The measured doses were compared to evaluate the differences in the amount of skin dose delivered by using these techniques. The results indicate that, on average, the patients treated with spot scanning proton beams received lower skin doses by an amount of 11.8% +/- 0.3% than did the patients treated with passively scattered proton beams. That difference could amount to 4 CGE per field for a prescribed dose of 76 CGE in 38 fractions treated with two equally weighted parallel opposed fields. PMID- 19550004 TI - Dose to water versus dose to medium in proton beam therapy. AB - Dose in radiation therapy is traditionally reported as the water-equivalent dose, or dose to water. Monte Carlo dose calculations report dose to medium and thus a methodology is needed to convert dose to medium into dose to water (or vice versa) for comparison of Monte Carlo results with results from planning systems. This paper describes the development of a formalism to convert dose to medium into dose to water for proton fields when simulating the dose with Monte Carlo techniques. The conversion is based on relative stopping power but also considers energy transferred via nuclear interactions. The influence of different interaction mechanisms of proton beams (electromagnetic versus nuclear) is demonstrated. Further, an approximate method for converting doses retroactively is presented. Based on the outlined formalism, five proton therapy patients with a total of 33 fields were analyzed. Dose distributions, dose volume histograms and absolute doses to assess the clinical significance of differences between dose to medium and dose to water are presented. We found that the difference between the two dose reporting definitions can be up to 10% for high CT numbers if analyzing the mean dose to the target. The difference is clinically insignificant for soft tissues. For the structures analyzed, the mean dose to water could be converted to dose to medium by applying a correction factor increasing linearly with increasing average CT number in the volume. We determined that an approximate conversion method, done retroactively with an energy-independent stopping power ratio and without considering nuclear interaction events separately (as compared to on-the-fly conversion during simulation), is sufficiently accurate to compute mean doses. It is insufficient, however, when analyzing the beam range. For proton beams stopping in bony anatomy, the predicted beam range can differ by 2-3 mm when comparing dose to tissue and dose to water. PMID- 19550005 TI - Some considerations concerning volume-modulated arc therapy: a stepping stone towards a general theory. AB - In this paper it is formally shown that the dynamic multileaf collimator (MLC) IMRT delivery technique remains valid if the MLC is supported on a 1D moving platform. It is also shown that, in such circumstances, it is always time preferable to deliver overlapping modulating fields as a single swept field rather than as separate fields. The most general formulism is presented and then related to simpler equations in limiting cases. The paper explains in detail how a 'small-arc approximation' can be invoked to relate the 1D linear theory to the MLC-on-moving-platform-(gantry) delivery technique involving rotation therapy and known as volume-modulated arc therapy (VMAT). It is explained how volume modulated arc therapy delivered with open unmodulated fields and which can deliver conformal dose distributions can be interpreted as an IMRT delivery. The (Elekta adopted) term VMAT will be used in a generic sense to include a similar (Varian) method known as RapidArc. Approximate expressions are derived for the 'amount of modulation' possible in a VMAT delivery. This paper does not discuss the actual VMAT planning but gives an insight at a deep level into VMAT delivery. No universal theory of VMAT is known in the sense that there is no theory that can predict precisely the performance of a VMAT delivery in terms of the free parameters available (variable gantry speed, variable fluence-delivery rate, set of MLC shapes, MLC orientation, number of arcs, coplanarity versus non coplanarity, etc). This is in stark contrast to the situation with several other IMRT delivery techniques where such theoretical analyses are known. In this paper we do not provide such a theory; the material presented is a stepping stone on the path towards this. PMID- 19550006 TI - Nanoscale roughness contact in a slider-disk interface. AB - The nanoscale roughness contact between molecularly smooth surfaces of a slider disk interface in a hard disk drive is analyzed, and the lubricant behavior at very high shear rate is presented. A new contact model is developed to study the nanoscale roughness contact behavior by classifying various forms of contact into slider-lubricant contact, slider-disk elastic contact and plastic contact. The contact pressure and the contact probabilities of the three types of contact are investigated. The new contact model is employed to explain and provide insight to an interesting experimental result found in a thermal protrusion slider. The protrusion budget for head surfing in the lubricant, which is the ideal state for contact recording, is also discussed. PMID- 19550007 TI - Intracellular nanomanipulation by a photonic-force microscope with real-time acquisition of a 3D stiffness matrix. AB - A traditional photonic-force microscope (PFM) results in huge sets of data, which requires tedious numerical analysis. In this paper, we propose instead an analog signal processor to attain real-time capabilities while retaining the richness of the traditional PFM data. Our system is devoted to intracellular measurements and is fully interactive through the use of a haptic joystick. Using our specialized analog hardware along with a dedicated algorithm, we can extract the full 3D stiffness matrix of the optical trap in real time, including the off-diagonal cross-terms. Our system is also capable of simultaneously recording data for subsequent offline analysis. This allows us to check that a good correlation exists between the classical analysis of stiffness and our real-time measurements. We monitor the PFM beads using an optical microscope. The force feedback mechanism of the haptic joystick helps us in interactively guiding the bead inside living cells and collecting information from its (possibly anisotropic) environment. The instantaneous stiffness measurements are also displayed in real time on a graphical user interface. The whole system has been built and is operational; here we present early results that confirm the consistency of the real-time measurements with offline computations. PMID- 19550008 TI - Catanionic-surfactant-controlled morphosynthesis and gas-sensing properties of corundum-type In(2)O(3). AB - Metastable corundum-type In(2)O(3) hierarchical structures with various morphologies inherited InOOH nanostructures, which were generated via a catanionic-surfactant-assisted solvothermal process. The products were characterized in detail by x-ray powder diffractometry (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and HRTEM. These superstructures were built from one-dimensional components. The one-dimensional preferential growth is explained in terms of the crystallographic features of InOOH. The effects of reaction parameters on the formation of nanostructures are also discussed. It is demonstrated that both laurylamine and oleic acid play an important role in the generation of InOOH hierarchical structures. Gas sensors composed of In(2)O(3) hierarchical nanostructures show a good response to toxic chlorophenol at 280 degrees C. PMID- 19550009 TI - Resistance switching in a SiC nanowire/Au nanoparticle network. AB - Resistance switching in a semiconductor nanowire/metal nanoparticle system is demonstrated. SiC nanowires grown on a Si substrate and decorated with Au nanoparticles are measured using W microprobes in a scanning electron microscope, where one probe is grounded and the other is biased. HIGH and LOW states can be toggled by applying a negative or positive pulse voltage. The switching mechanism is attributed to a charge transfer between the SiC nanowires and the Au nanoparticles. PMID- 19550010 TI - Synthesis and characterization of Au-attached single-walled carbon nanotube bundles. AB - We synthesized suspended single-walled carbon nanotubes (SWNTs) by the thermal chemical vapor deposition method and functionalized them with Au nanoparticles (NPs). We used 3-(aminopropyl)triethoxysilane as a linker and controlled the Au NP density on the SWNT surface by changing the reaction time. In the Raman scattering spectra of the Au-functionalized SWNTs, an enhanced peak frequency and peak intensity were observed in the non-resonant region. A significant enhancement of the metallic character in the high frequency region was also observed, especially when we used a 633 nm laser. By measuring the electric properties using a standard field effect transistor configuration, we found that charge transfer occurred during the functionalization processes. It is expected that the charge transfer related optical enhancement may affect the observed change in the Raman profiles. PMID- 19550011 TI - The structural and electronic properties of chiral SiC nanotubes: a hybrid density functional study. AB - Hybrid density functional theory was employed in investigating the structural and electronic properties of 14 chiral and 3 armchair SiC nanotubes (SiCNTs). The role of the tube diameter, as well as that of the chiral angle theta, was studied in detail by considering nanotubes of diameters varying from 2 to 9 A and chiral angles theta varying between 7 degrees and 30 degrees. The study revealed that all the investigated SiCNTs are semiconductors with a broad spectrum of bandgap values ranging from 0.2 to 2.9 eV and that the structural stability of the nanotubes increases with diameter. By analyzing the behavior of the molecular orbitals, an explanation of the mechanism by which theta affects the determination of such values is put forward. PMID- 19550012 TI - Micellar nanotubes and AAO nanopores decorated with nanoparticles. AB - We demonstrated that anodized aluminum oxide (AAO) enables the fabrication of micellar nanotubes and nanopores decorated with nanoparticles. Block copolymer micelles containing precursors of nanoparticles were coated on the nanopores of AAO, from which we were able to select nanotubes containing nanoparticles or nanopores decorated with nanoparticles by removing the AAO template or the copolymer. Upon removal of the AAO, the micellar nanotubes with nanoparticles were produced, whereas the nanopores of AAO were decorated with nanoparticles by eliminating the copolymer. Since a variety of nanoparticles can be synthesized in addition to full control over the size and spacing of nanoparticles by the copolymer micellar approach, the methodology demonstrated here can allow fabricating functional nanotubes or nanopores with a selection of the type and size of nanoparticles. PMID- 19550013 TI - Electrochemical and surface plasmon resonance characterization of beta cyclodextrin-based self-assembled monolayers and evaluation of their inclusion complexes with glucocorticoids. AB - This paper describes the characterization of a self-assembled beta-cyclodextrin (beta-CD)-derivative monolayer (beta-CD-SAM) on a gold surface and the study of their inclusion complexes with glucocorticoids. To this aim the arrangement of a self-assembled beta-cyclodextrin-derivative monolayer on a gold surface was monitored in situ by means of surface plasmon resonance (SPR) spectroscopy and double-layer capacitance measurements. Film thickness and dielectric constant were evaluated for a monolayer of beta-CD using one-color-approach SPR. The selectivity of the beta-CD host surface was verified by using electroactive species permeable and impermeable in the beta-CD cavity. The redox probe was selected according to its capacity to permeate the beta-CD monolayer and its electrochemical behavior. In order to evaluate the feasibility of an inclusion complex between beta-CD-SAM with some steroids such as cortisol and cortisone, voltammetric experiments in the presence of the redox probes as molecules competitive with the steroids have been performed. The formation constant of the surface host-guest by beta-CD-SAM and the steroids under study was calculated. PMID- 19550014 TI - The synthesis of high coercivity cobalt-in-carbon nanotube hybrid structures and their optical limiting properties. AB - Magnetic heterostructures with carbon nanotubes having multiple functionalities are fascinating materials which can be manipulated by means of an external magnetic field. In this paper we report our investigations on the synthesis and optical limiting properties of pristine cobalt nanotubes and high coercivity cobalt-in-carbon nanotubes (a new nanosystem where carbon nanotubes are filled with cobalt nanotubes). A general mobility assisted growth mechanism for the formation of one-dimensional nanostructures inside nanopores is verified in the case of carbon nanotubes. The open-aperture z-scan technique is employed for the optical limiting measurements in which nanosecond laser pulses at 532 nm have been used for optical excitation. Compared to the benchmark pristine carbon nanotubes these materials show an enhanced nonlinear optical absorption, and the nonlinear optical parameters calculated from the data show that these materials are efficient optical limiters. To the best of our knowledge this is the first report where the optical limiting properties of metal nanotubes are compared to those of carbon nanotubes. PMID- 19550015 TI - Fe oxidation versus Pt segregation in FePt nanoparticles and thin films. AB - Metallic nanoparticles containing 3d elements are generally susceptible to oxidation leading to a deterioration of desired properties. Here, the oxidation behavior of differently sized FePt nanoparticles is experimentally studied by x ray photoelectron spectroscopy (XPS) and compared to a FePt reference film. For all as-prepared metallic samples the common features are the formation of Fe(3+), becoming detectable for exposures to pure oxygen above 10(6) langmuir whereas under identical conditions the Pt(0) signal is conserved. Most notably, these features are independent of particle size. Annealing at 650 degrees C, however, affects small and large FePt particles differently. While large particles as well as the reference film show a 100-1000 times enhanced resistance against oxidation, small FePt particles (diameter 5 nm) exhibit no such enhancement due to the thermal treatment. Additional XPS intensity analysis in combination with model calculations leads to an explanation of this observation in terms of Pt segregating to the surface. In large particles and films the thickness of the resulting Pt layer is sufficient to strongly impede oxidation, while in small particles this layer is incomplete and no longer provides protection against oxidation. PMID- 19550016 TI - Influence of the macroscopic shape of the tip on the contrast in scanning polarization force microscopy images. AB - We demonstrate that a quantitative analysis of the contrast obtained in electrostatic force microscopy images that probe the dielectric response of the sample (scanning polarization force microscopy (SPFM)) requires numerical simulations that take into account both the macroscopic shape of the tip and the nanoscopic tip apex. To simulate the SPFM contrast, we have used the generalized image charge method (GICM), which is able to accurately deal with distances between a few nanometers and several microns, thus involving more than three orders of magnitude. Our numerical simulations show that the macroscopic shape of the tip accounts for most of the SPFM contrast. Moreover, we find a quasi-linear relation between the working tip-sample distance and the contrast for tip radii between 50 and 200 nm. Our calculations are compared with experimental measurements of the contrast between a thermally grown silicon oxide sample and a few-layer graphene film transferred onto it. PMID- 19550018 TI - The formation of TiO(2) nanowires directly from nanoparticles. AB - TiO(2) nanowires were fabricated by annealing TiO(2) nanoparticles on silicon substrate at 1000 degrees C in air. When a polystyrene nanosphere monolayer was used as a template to separate the TiO(2) nanoparticles, they could more easily react with the silicon substrate to form Ti(5)Si(3). The TiO(2) nanowires were formed upon further oxidation of Ti(5)Si(3). The diameters and lengths of TiO(2) nanowires were 30-80 nm and 1-3 microm, respectively. The nanowires had a rutile structure with the growth direction [112]. It is believed that the formation of TiO(2) nanowires involved a precipitation process in the mixture of SiO(2) and TiO(2). The nanowires show different photoluminescence behavior from that of the powder. PMID- 19550017 TI - In situ TEM observation of the growth and decomposition of monoclinic W(18)O(49) nanowires. AB - The growth of monoclinic W(18)O(49) nanowires by heat treatment of a tungsten filament at approximately 873 K and the decomposition of these nanowires under 200 keV electron irradiation at approximately 1023 K have been investigated using in situ transmission electron microscopy (TEM). In situ TEM observation of the growth confirmed the vapor-solid growth mechanism of the monoclinic W(18)O(49) nanowires. In situ irradiation experiments revealed the formation of metallic bcc tungsten from monoclinic W(18)O(49) nanowires under 200 keV electron irradiation. PMID- 19550019 TI - Size and density control of In droplets at near room temperatures. AB - We report on the ability to control the size and density of In droplets on GaAs(100) substrates at near room temperatures using solid source molecular beam epitaxy. We specifically demonstrate the height, diameter and density control of In droplets as functions of substrate temperature (T(sub)) and monolayer (ML) coverage. For a range of density (approximately 10(9)-10(10) cm(-2)), the growth window is revealed to be between 20 and 70 degrees C. For a fixed ML coverage, the size and density of droplets can be controlled by controlling the T(sub). For a fixed T(sub), by controlling the ML coverage, droplet size and density can be controlled. Even at near room temperatures (20-70 degrees C), In atoms are extremely sensitive to surface diffusion and this enables the control of the size and density of droplets. This study provides an aid to understanding the formation of In droplets at near room temperatures and can find applications in the formation of quantum structures and/or nanostructures based on droplet epitaxy. PMID- 19550020 TI - Room temperature synthesized rutile TiO(2) nanoparticles induced by laser ablation in liquid and their photocatalytic activity. AB - TiO(2) nanoparticles were prepared by one-step pulsed laser ablation of a titanium target immersed in a poly-(vinylpyrrolidone) solution at room temperature. The products were systematically characterized by x-ray diffraction (XRD), transmission electron microscopy (TEM), Raman spectroscopy and x-ray photoelectron spectroscopy (XPS). The results indicated that the rutile TiO(2) nanocrystalline particles were one-step synthesized at room temperature and the mean size in diameter is about 50 nm with a narrow size distribution. A probable formation process was proposed on the basis of the microstructure and the instantaneous plasma plume induced by the laser. Photocatalytic activity was monitored by degradation of a methylene blue solution. The as-prepared rutile TiO(2) nanoparticles demonstrate a good photocatalytic performance. This work shows that pulsed laser ablation in liquid media is a good method to synthesize some nanosized materials which are difficult to produce by other conventional methods. PMID- 19550021 TI - The controlled growth of single metallic and conducting polymer nanowires via gate-assisted electrochemical deposition. AB - The fabrication of nanowires with well-controlled lengths and diameters is the basis of the application of one-dimensional nanostructures in more sophisticated electronic and biomolecular device systems. A wide variety of materials, including metals and conducting polymers, have been utilized in nanowire arrays as building blocks for chemical or biomolecular sensors. Thus far, the cheapest and most effective way of nanowire synthesis is electrochemical deposition. In this work, we investigate a new method of electrochemical deposition using two dimensional electric fields instead of the conventional one-directional electric field between working electrodes. Reproducible fabrication of metallic (palladium) and conducting polymer (polypyrrole) single nanowires with diameters down to 30-50 nm is achieved by application of a vertical gate electric field in addition to the lateral one between the two working electrodes. Diameters and lengths of the nanowires can be easily controlled by varying the dimensions of the nanochannels in which the nanowires are grown. A good ohmic contact between the nanowire and gold electrodes is also obtained, indicating the feasibility of electronic devices based on the single nanowires synthesized via this method. In conjunction with experimental findings of nanowire growth mechanism under two dimensional electric field, molecular dynamic simulations are employed to further understand the deposition process. This improved electrochemical deposition is applicable for controlled and simple fabrication of a wide range of metallic and conducting polymeric nanowires with small diameters. PMID- 19550022 TI - The geometric effect and programming current reduction in cylindrical-shaped phase change memory. AB - This study conducts a three-dimensional electro-thermal time-domain simulation for numerical analysis of cylindrical-shaped phase change memories (PCMs). The influence of chalcogenide material, germanium antimony telluride (GeSbTe or GST), structure on PCM operation is explored. GST with vertical structure exhibits promising characteristics. The bottom electrode contact (BEC) is advanced to improve the operation of PCMs, where a 25% reduction of the required programming current is achieved at a cost of 26% reduced resistance ratio. The position of the BEC is then shifted to further improve the performance of PCMs. The required programming current is reduced by a factor of 11, where the resistance ratio is only decreased by 6.9%. However, the PCMs with a larger shift of BEC are sensitive to process variation. To design PCMs with less than 10% programming current variation, PCMs with shifted BEC, where the shifted distance is equal to 1.5 times the BEC's radius, is worth considering. This study quantitatively estimates the structure effect on the phase transition of PCMs and physically provides an insight into the design and technology of PCMs. PMID- 19550023 TI - Diamagnetically stabilized levitation control of an intraluminal magnetic capsule. AB - Controlled navigation promotes full utilization of capsule endoscopy for reliable real-time diagnosis in the gastrointestinal (GI) tract, but intermittent natural peristalsis can disturb the navigational control, destabilize the capsule and take it out of levitation. The focus of the present work was to develop an economical and effective real-time magnetic capsule-guiding system that can operate in the presence of naturally existing peristalsis while retaining navigational control. A real-size magnetic navigation system that can handle peristaltic forces of up to 1.5 N was designed utilizing the computer-aided design (CAD) system Maxwell 3D (Ansoft, Pittsburg, PA) and was verified using a small-size physical experimental setup. The proposed system contains a pair of 50 cm diameter, 10,000-turn copper electromagnets with a 10 cm x 10 cm ferrous core driven by currents of up to 300 A and can successfully maintain position control over the levitating capsule during peristalsis. The addition of bismuth diamagnetic casing for stabilizing the levitating capsule was also studied. A modeled magnetic field around the diamagnetically cased permanent magnet was shown to be redistributed aligning its interaction with the external electromagnets, thus stabilizing the levitating capsule. In summary, a custom designed diamagnetically facilitated capsule navigation system can successfully steer an intraluminal magnet-carrying capsule. PMID- 19550024 TI - Inter-electrode delay estimators for electrohysterographic propagation analysis. AB - Premature birth is a major cause of mortality and permanent dysfunctions. Several parameters derived from single channel electrohysterographic (EHG) signals have been considered to determine contractions leading to preterm delivery. The results are promising, but improvements are needed. As effective uterine contractions result from a proper action potential propagation, in this paper we focus on the propagation properties of EHG signals, which can be predictive of preterm delivery. Two standard delay estimators, namely maximization of the cross correlation function and spectral matching, are adapted and implemented for the assessment of inter-electrode delays of propagating EHG signals. The accuracy of the considered standard estimators might be hampered by a poor inter-channel correlation. An improved dedicated approach is therefore proposed. By simultaneous adaptive estimation of the volume conductor transfer function and the delay, a dedicated method is conceived for improving the inter-channel signal similarity during delay calculation. Furthermore, it provides delay estimates without resolution limits and it is suitable for low sampling rates, which are appropriate for EHG recording. The three estimators were evaluated on EHG signals recorded on seven women. The dedicated approach provided more accurate estimates due to a 22% improvement of the initial average inter-channel correlation. PMID- 19550025 TI - Objective selection of EEG late potentials through residual dependence estimation of independent components. AB - This paper presents a novel method to objectively select electroencephalographic (EEG) cortical sources estimated by independent component analysis (ICA) in event related potential (ERP) studies. A proximity measure based on mutual information is employed to estimate residual dependences of the components that are then hierarchically clustered based on these residual dependences. Next, the properties of each group of components are evaluated at each level of the hierarchical tree by two indices that aim to assess both cluster tightness and physiological reliability through a template matching process. These two indices are combined in three different approaches to bring to light the hierarchical structure of the cluster organizations. Our method is tested on a set of experiments with the purpose of enhancing late positive ERPs elicited by emotional picture stimuli. Results suggest that the best way to look for physiologically plausible late positive potential (LPP) sources is to explore in depth the tightness of those clusters that, taken together, best resemble the template. According to our results, after brain sources clustering, LPPs are always identified more accurately than from ensemble-averaged raw data. Since the late components of an ERP involve the same associative areas, regardless of the modality of stimulation or specific tasks administered, the proposed method can be simply adapted to other ERP studies, and extended from psychophysiological studies to pathological or sport training evaluation support. PMID- 19550026 TI - Complexity analysis of EEG in patients with schizophrenia using fractal dimension. AB - We computed Higuchi's fractal dimension (FD) of resting, eyes closed EEG recorded from 30 scalp locations in 18 male neuroleptic-naive, recent-onset schizophrenia (NRS) subjects and 15 male healthy control (HC) subjects, who were group-matched for age. Schizophrenia patients showed a diffuse reduction of FD except in the bilateral temporal and occipital regions, with the reduction being most prominent bifrontally. The positive symptom (PS) schizophrenia subjects showed FD values similar to or even higher than HC in the bilateral temporo-occipital regions, along with a co-existent bifrontal FD reduction as noted in the overall sample of NRS. In contrast, this increase in FD values in the bilateral temporo-occipital region was absent in the negative symptom (NS) subgroup. The regional differences in complexity suggested by these findings may reflect the aberrant brain dynamics underlying the pathophysiology of schizophrenia and its symptom dimensions. Higuchi's method of measuring FD directly in the time domain provides an alternative for the more computationally intensive nonlinear methods of estimating EEG complexity. PMID- 19550027 TI - The application of empirical mode decomposition for the enhancement of cardiotocograph signals. AB - Cardiotocograph (CTG) is widely used in everyday clinical practice for fetal surveillance, where it is used to record fetal heart rate (FHR) and uterine activity (UA). These two biosignals can be used for antepartum and intrapartum fetal monitoring and are, in fact, nonlinear and non-stationary. CTG recordings are often corrupted by artifacts such as missing beats in FHR, high-frequency noise in FHR and UA signals. In this paper, an empirical mode decomposition (EMD) method is applied on CTG signals. A recursive algorithm is first utilized to eliminate missing beats. High-frequency noise is reduced using EMD followed by the partial reconstruction (PAR) method, where the noise order is identified by a statistical method. The obtained signal enhancement from the proposed method is validated by comparing the resulting traces with the output obtained by applying classical signal processing methods such as Butterworth low-pass filtering, linear interpolation and a moving average filter on 12 CTG signals. Three obstetricians evaluated all 12 sets of traces and rated the proposed method, on average, 3.8 out of 5 on a scale of 1(lowest) to 5 (highest). PMID- 19550028 TI - mGluR5 is a central regulator of synaptic function and plasticity in the developing mouse barrel cortex. PMID- 19550029 TI - Snakes and ladders: the ups and downs of animal segmentation. PMID- 19550030 TI - Insufficient regulatory supervision prior to release of genetically modified crops for commercial cultivation in India. PMID- 19550031 TI - Green fluorescent protein: a molecular lantern that illuminates the cellular interior. PMID- 19550032 TI - Why Darwin rejected intelligent design. PMID- 19550033 TI - The history of vitamin C research in India. PMID- 19550034 TI - What history tells us XVII. Conrad Waddington and the nature of life. PMID- 19550035 TI - Inactivation of a transgene due to transposition of insertion sequence (IS136) of Agrobacterium tumefaciens. AB - Agrobacterium strains harbour insertion sequences, which are known to transpose into genomes as well as into Ti plasmids. In this study we report the inactivation of a transgene due to transposition of the A. tumefaciens insertion sequence IS136. The transposition was discovered following transformation of plant tissues, although the fidelity of the binary vector was confirmed following transformation into Agrobacterium. Such transpositions are rare but can occur and it is thus important to check the fidelity of the binary vector at different times of Agrobacterium growth in order to avoid failure in achieving transgene expression. PMID- 19550036 TI - Functional role of EF-hands 3 and 4 in membrane-binding of KChIP1. AB - The aim of the present study is to explore whether membrane targeting of K+ channel-interacting protein 1 (KChIP1) is associated with its EF-hand motifs and varies with specific phospholipids. Truncated KChIP1, in which the EFhands 3 and 4 were deleted, retained the alpha-helix structure, indicating that the N terminal half of KChIP1 could fold appropriately. Compared with wild-type KChIP1, truncated KChIP1 exhibited lower lipid-binding capability. Compared with wild type KChIP1, increasing membrane permeability by the use of digitonin caused a marked loss of truncated KChIP1, suggesting that intact EF-hands 3 and 4 were crucial for the anchorage of KChIP1 on membrane. KChIP1 showed a higher binding capability with phosphatidylserine (PS) than truncated KChIP1. Unlike that of truncated KChIP1, the binding of wild-type KChIP1 with membrane was enhanced by increasing the PS content. Moreover, the binding of KChIP1 with phospholipid vesicles induced a change in the structure of KChIP1 in the presence of PS. Taken together, our data suggest that EF-hands 3 and 4 of KChIP1 are functionally involved in a specific association with PS on the membrane. PMID- 19550037 TI - Identification of binding peptides of the ADAM15 disintegrin domain using phage display. AB - ADAM15 plays an important role in tumour development by interacting with integrins. In this study, we investigated the target peptides of the ADAM15 disintegrin domain. First, we successfully produced the recombinant human ADAM15 disintegrin domain (RADD) that could inhibit melanoma cell adhesion by using Escherichia coli. Second, four specific binding peptides (peptides A, B, C, and D) were selected using a phage display 12-mer peptide library. The screening protocol involved 4 rounds of positive panning on RADD and 2 rounds of subtractive selection with streptavidin. By using the BLAST software and a relevant protein database, integrin alpha v beta 3 was found to be homologous to peptide A. Synthetic peptide A had a highly inhibitory effect on RADD-integrin alpha v beta 3 binding. The results demonstrate the potential application of short peptides for disrupting high-affinity ADAM-integrin interactions. PMID- 19550038 TI - Selection of scFvs specific for the HepG2 cell line using ribosome display. AB - The aim of this study was to construct a ribosome display library of single chain variable fragments (scFvs) associated with hepatocarcinoma and screen such a library for hepatocarcinoma-binding scFvs. mRNA was isolated from the spleens of mice immunized with hepatocellular carcinoma cell line HepG2. Heavy and k chain genes (VH and k) were amplified separately by RT-PCR, and an anti-HepG2 VH/k chain ribosome display library was constructed by assembling VH and k into the VH/k chain with a specially constructed linker by SOE-PCR. The VH/k chain library was transcribed and translated in vitro using a rabbit reticulocyte lysate system. In order to isolate specific scFvs, recognizing HepG2 negative selection on a normal hepatocyte line WRL-68 was carried out before three rounds of positive selection on HepG2. After three rounds of panning, cell enzyme-linked immunosorbent assay (ELISA) showed that one of the scFvs had high affinity for the HepG2 cell and lower affinity for the WRL-68 cell. In this study, we successfully constructed a native ribosome display library. Such a library would prove useful for direct intact cell panning using ribosome display technology. The selected scFv had a potential value for hepatocarcinoma treatment. PMID- 19550040 TI - Interaction of 18-residue peptides derived from amphipathic helical segments of globular proteins with model membranes. AB - We investigated the interaction of six 18-residue peptides derived from amphipathic helical segments of globular proteins with model membranes. The net charge of the peptides at neutral pH varies from -1 to +6. Circular dichroism spectra indicate that peptides with a high net positive charge tend to fold into a helical conformation in the presence of negatively charged lipid vesicles. In helical conformation, their average hydrophobic moment and hydrophobicity would render them surface-active. The composition of amino acids on the polar face of the helix in the peptides is considerably different. The peptides show variations in their ability to permeabilise zwitterionic and anionic lipid vesicles. Whereas increased net positive charge favours greater permeabilisation, the distribution of charged residues in the polar face also plays a role in determining membrane activity. The distribution of amino acids in the polar face of the helix in the peptides that were investigated do not fall into the canonical classes described. Amphipathic helices, which are part of proteins, with a pattern of amino acid distribution different from those observed in class L, A and others, could help in providing newer insights into peptide-membrane interactions. PMID- 19550041 TI - Physical mapping, expression analysis and polymorphism survey of resistance gene analogues on chromosome 11 of rice. AB - Rice is the first cereal genome with a finished sequence and a model crop that has important syntenic relationships with other cereal species. The objectives of our study were to identify resistance gene analogue (RGA) sequences from chromosome 11 of rice, understand their expression in other cereals and dicots by in silico analysis, determine their presence on other rice chromosomes, and evaluate the extent of polymorphism and actual expression in a set of rice genotypes. A total of 195 RGAs were predicted and physically localised. Of these, 91.79% expressed in rice, and 51.28% expressed in wheat, which was the highest among other cereals. Among monocots, sugarcane showed the highest (78.92%) expression, while among dicots, RGAs were maximally expressed in Arabidopsis (11.79%). Interestingly, two of the chromosome 11-specific RGAs were found to be expressing in all the organisms studied. Eighty RGAs of chromosome 11 had significant homology with chromosome 12, which was the maximum among all the rice chromosomes. Thirty-one per cent of the RGAs used in polymerase chain reaction (PCR) amplification showed polymorphism in a set of rice genotypes. Actual gene expression analysis revealed post-inoculation induction of one RGA in the rice line IRBB-4 carrying the bacterial blight resistance gene Xa-4. Our results have implications for the development of sequence-based markers and functional validation of specific RGAs in rice. PMID- 19550039 TI - Cloning and sequence analysis of putative type II fatty acid synthase genes from Arachis hypogaea L. AB - The cultivated peanut is a valuable source of dietary oil and ranks fifth among the world oil crops. Plant fatty acid biosynthesis is catalysed by type II fatty acid synthase (FAS) in plastids and mitochondria. By constructing a full-length cDNA library derived from immature peanut seeds and homology-based cloning, candidate genes of acyl carrier protein (ACP), malonyl-CoA:ACP transacylase, beta ketoacyl-ACP synthase (I, II, III), beta-ketoacyl-ACP reductase, beta-hydroxyacyl ACP dehydrase and enoyl-ACP reductase were isolated. Sequence alignments revealed that primary structures of type II FAS enzymes were highly conserved in higher plants and the catalytic residues were strictly conserved in Escherichia coli and higher plants. Homologue numbers of each type II FAS gene expressing in developing peanut seeds varied from 1 in KASII, KASIII and HD to 5 in ENR. The number of single-nucleotide polymorphisms (SNPs) was quite different in each gene. Peanut type II FAS genes were predicted to target plastids except ACP2 and ACP3. The results suggested that peanut may contain two type II FAS systems in plastids and mitochondria. The type II FAS enzymes in higher plants may have similar functions as those in E. coli. PMID- 19550042 TI - Variations in morphological and life-history traits under extreme temperatures in Drosophila ananassae. AB - Using half-sib analysis, we analysed the consequences of extreme rearing temperatures on genetic and phenotypic variations in the morphological and life history traits of Drosophila ananassae. Paternal half-sib covariance contains a relatively small proportion of the epistatic variance and lacks the dominance variance and variance due to maternal effect, which provides more reliable estimates of additive genetic variance. Experiments were performed on a mass culture population of D. ananassae collected from Kanniyakumari (India). Two extremely stressful temperatures (18 degree C and 32 degree C) and one standard temperature (25 degree C) were used to examine the effect of stressful and non stressful environments on the morphological and life-history traits in males and females. Mean values of various morphological traits differed signifi cantly among different temperature regimens in both males and females. Rearing at 18 degree C and 32 degree C resulted in decreased thorax length, wing-to-thorax (w/t) ratio, sternopleural bristle number, ovariole number, sex comb-tooth number and testis length. Phenotypic variances increased under stressful temperatures in comparison with non-stressful temperatures. Heritability and evolvability based on among-sires (males), among-dams (females), and the sum of the two components (sire + dam) showed higher values at both the stressful temperatures than at the non-stressful temperature. These differences reflect changes in additive genetic variance. Viability was greater at the high than the low extreme temperature. As viability is an indicator of stress, we can assume that stress was greater at 18 degree C than at 32 degree C in D. ananassae. The genetic variations for all the quantitative and life-history traits were higher at low temperature. Variation in sexual traits was more pronounced as compared with other morphometric traits, which shows that sexual traits are more prone to thermal stress. Our results agree with the hypothesis that genetic variation is increased in stressful environments. PMID- 19550043 TI - Effect of directional selection for body size on fluctuating asymmetry in certain morphological traits in Drosophila ananassae. AB - Variation in the subtle differences between the right and left sides of bilateral characters or fluctuating asymmetry (FA) has been considered as an indicator of an organism's ability to cope with genetic and environmental stresses during development. However, due to inconsistency in the results of empirical studies, the relationship between FA and stress has been the subject of intense debate. In this study, we investigated whether stress caused by artificial bidirectional selection for body size has any effect on the levels of FA of different morphological traits in Drosophila ananassae. The realised heritability (h2) was higher in low-line females and high-line males, which suggests an asymmetrical response to selection for body size. Further, the levels of FA were compared across 10 generations of selection in different selection lines in both sexes for sternopleural bristle number, wing length, wing-to-thorax ratio, sex combtooth number and ovariole number. The levels of FA differed significantly among generations and selection lines but did not change markedly with directional selection. However, the levels of FA were higher in the G10 generation (at the end of selection) than G0 (at the start of selection) but lower than the G5 generation in different selection lines, suggesting that the levels of FA are not affected by the inbreeding generated during the course of selection. Also, the levels of FA in the hybrids of high and low lines were signifi cantly lower than the parental selection lines, suggesting that FA is influenced by hybridisation. These results are discussed in the framework of the literature available on FA and its relationship with stress. PMID- 19550044 TI - Bioluminescence emissions of the firefly Luciola praeusta Kiesenwetter 1874 (Coleoptera : Lampyridae : Luciolinae). AB - We recorded the in vivo emission and time-resolved spectra of the firefly Luciola praeusta Kiesenwetter 1874 (Coleoptera : Lampyridae : Luciolinae). The emission spectrum shows that the full width at half maximum (FWHM) value for this particular species is 55 nm, which is significantly narrower than the in vivo half-widths reported till now. The time-resolved spectrum reveals that a flash of about 100 ms duration is, in fact, composed of a number of microsecond pulses. This suggests that the speed of the enzyme-catalysed chemiluminescence reaction in the firefly for the emission of light is much faster than was previously believed. PMID- 19550045 TI - Rapid aggregation and assembly in aqueous solution of A beta (25-35) peptide. AB - The highly toxic A beta (25-35) is a peculiar peptide that differs from all the other commonly studied beta-amyloid peptides because of its extremely rapid aggregation properties and enhanced neurotoxicity. We investigated A beta (25-35) aggregation in H2O at pH 3.0 and at pH 7.4 by means of in-solution analyses. Adopting UV spectroscopy, Congo red spectrophotometry and thioflavin T fluorimetry, we were able to quantify, in water, the very fast assembling time necessary for A beta (25-35) to form stable insoluble aggregates and their ability to seed or not seed fibril growth. Our quantitative results, which confirm a very rapid assembly leading to stable insoluble aggregates of A beta (25-35) only when incubated at pH 7.4, might be helpful for designing novel aggregation inhibitors and to shed light on the in vivo environment in which fibril formation takes place. PMID- 19550046 TI - Retinoblastoma protein: a central processing unit. AB - The retinoblastoma protein (pRb) is one of the key cell-cycle regulating proteins and its inactivation leads to neoplastic transformation and carcinogenesis. This protein regulates critical G1 -to-S phase transition through interaction with the E2F family of cell-cycle transcription factors repressing transcription of genes required for this cell-cycle check-point transition. Its activity is regulated through network sensing intracellular and extracellular signals which block or permit phosphorylation (inactivation) of the Rb protein. Mechanisms of Rb dependent cell-cycle control have been widely studied over the past couple of decades. However, recently it was found that pRb also regulates apoptosis through the same interaction with E2F transcription factors and that Rb-E2F complexes play a role in regulating the transcription of genes involved in differentiation and development. PMID- 19550047 TI - MMP-1 polymorphism and its relationship to pathological processes. AB - Matrix metalloproteinases (MMPs) are a family of zinc (Zn)-dependent endopeptidases that are collectively capable of cleaving virtually all extracellular matrix (ECM) substrates and play an important role in diverse physiological and pathological processes. The activity of MMPs is regulated at multiple levels. The transcriptional regulation of MMP appears to represent the key step in MMP regulation. There are diverse types of MMPs that differ structural and functionally. MMP-1 is the most ubiquitously expressed interstitial collagenase and has a prominent role in initial cleavage of the ECM. The level of MMP-1 expression can be influenced by different single-nucleotide polymorphisms (SNPs) in the promoter region. A functional polymorphism at position -1607 has been shown to alter the transcriptional activity of MMP-1 and was associated with diverse pathological processes. The aim of our review was to discuss some topics related to MMP in physiological and pathological processes, with a focus on MMP-1 polymorphism. PMID- 19550049 TI - Compensation for research-related injury. PMID- 19550048 TI - Living with urea stress. AB - Intracellular organic osmolytes are present in certain organisms adapted to harsh environments. These osmolytes protect intracellular macromolecules against denaturing environmental stress. In contrast to the usually benign effects of most organic osmolytes, the waste product urea is a well-known perturbant of macromolecules. Although urea is a perturbing solute which inhibits enzyme activity and stability, it is employed by some species as a major osmolyte. The answer to this paradox was believed to be the discovery of protective osmolytes (methylamines). We review the current state of knowledge on the various ways of counteracting the harmful effects of urea in nature and the mechanisms for this. This review ends with the mechanistic idea that cellular salt (KCl/NaCl) plays a crucial role in counteracting the effects of urea, either by inducing required chaperones or methylamines, or by thermodynamic interactions with ureadestabilised proteins. We also propose future opportunities and challenges in the field. PMID- 19550051 TI - To be awake or not to be... PMID- 19550050 TI - Risk factors for diabetic retinopathy in rural India. PMID- 19550052 TI - Risk factors for diabetic retinopathy in self-reported rural population with diabetes. AB - BACKGROUND: Diabetes and its related microvascular complications like diabetic retinopathy (DR) are showing increased prevalence in India. However, the magnitude of DR in rural population with diabetes needs exploration. AIM: To estimate the prevalence and risk factors for the presence and severity of diabetic retinopathy in the self-reported rural population with diabetes. SETTINGS AND DESIGN: In a cross-sectional study, a total of 26,519 participants (age >or= 30 years) attended 198 diabetic retinopathy screening camps conducted in three southern districts of Tamilnadu, India, between February 2004 and April 2006. MATERIALS AND METHODS: All the participants underwent a dilated eye examination to detect DR by indirect ophthalmoscopy. Systemic and ocular risk factor estimation was done in a comprehensive examination. STATISTICAL ANALYSIS: Univariate and stepwise regression analyses were done to identify the independent risk factors associated with the presence and severity of retinopathy. RESULTS: The prevalence of diabetic retinopathy was 17.6% among the self-reported rural population with diabetes. The prevalence of referable (sight threatening) retinopathy was 5.3%. Risk factors associated with the development of any DR were male gender (OR= 1.37), longer duration of diabetes (per year, OR= 1.07), lean body mass index (OR= 1.30), higher systolic blood pressure (per 10 mm Hg, OR= 1.18), and insulin treatment (OR= 1.34; P P CONCLUSION: The study identified risk factors associated with DR in the rural population with diabetes. The results suggested that there was a need for formulating effective preventive strategies to minimize avoidable blindness due to diabetes, in rural areas. PMID- 19550053 TI - Quality of life of parents of children with newly diagnosed specific learning disability. AB - BACKGROUND: Poor school performance in children causes significant stress to parents. AIMS: To analyze the quality of life (QOL) of parents having a child with newly diagnosed specific learning disability (SpLD) and to evaluate the impact of clinical and socio-demographic characteristics on their QOL. DESIGN: Cross-sectional questionnaire-based study. SETTING: Learning disability clinic in tertiary care hospital. MATERIALS AND METHODS: From June 2006 to February 2007, 150 parents (either mother or father) of children consecutively diagnosed as having SpLD were enrolled. Parent's QOL was measured by the WHOQOL-100 instrument which is a generic instrument containing 25 facets of QOL organized in six domains. STATISTICAL ANALYSIS USED: Independent samples t-test, one-way analysis of variance, and multiple regression analysis were carried out for statistical significance. RESULTS: Mean age of parents was 42.6 years (SD 5.5); mothers to fathers ratio 1.3:1; and 19 (12.7%) were currently ill. Only four WHOQOL-100 domains (psychological > social relationships > environment > spiritual) and five WHOQOL-100 facets (leisure > feel > energy > esteem > sex) contributed significantly to their "overall" QOL. Female gender, being currently ill, being in paid work, and having a male child were characteristics that independently predicted a poor domain/facet QOL score. CONCLUSIONS: The present study has identified domains and facets that need to be addressed by counselors for improving overall QOL of these parents. Initiating these measures would also improve the home environment and help in the rehabilitation of children with SpLD. PMID- 19550054 TI - Clinical information in drug package inserts in India. AB - BACKGROUND: It is widely recognized that accurate and reliable product information is essential for the safe and effective use of medications. Pharmaceutical companies are the primary source of most drug information, including package inserts. Package inserts are printed leaflets accompanying marketed drug products and contain information approved by the regulatory agencies. Studies on package inserts in India, in 1996, had shown that crucial information was often missing and they lacked uniformity. AIM: To assess the presentation and completeness of clinically important information provided in the currently available package inserts in India. MATERIALS AND METHODS: Package inserts accompanying allopathic drug products marketed by pharmaceutical companies in India were collected. These package inserts were analyzed for the content of clinically important information in various sections. STATISTICAL ANALYSIS: The results were expressed as absolute numbers and percentages. RESULTS: Preliminary analyses revealed that most package inserts did contain information under headings, such as, therapeutic indications, contraindications, undesirable effects, etc., listed in the Drugs and Cosmetics Rules 1945. The findings indicated considerable improvement in package inserts since 1996. However, on critical evaluation it was revealed that clinically important information was not well presented and was often incomplete. Information with regard to pediatric and geriatric use was present in only 44% and 13% of the package inserts, respectively. Only five of the inserts had information on the most frequent adverse drug reactions associated with the drug. Also, information on interactions and overdosage was often missing. CONCLUSION: Although the package inserts appear to have improved over the past decade there is still a definite need to further refine the clinical information contained, to minimize the risks to patients. This could be brought about by self-regulation on the part of the industry as well as by updating the relevant guidelines in line with those of developed countries. PMID- 19550055 TI - Patient controlled sedation during central neuraxial anesthesia. AB - BACKGROUND: Patient controlled sedation (PCS) gives liberty to patients to choose the time of sedative administration to attain a desired level of comfort. AIMS: The PCS use was evaluated in patients during surgery under central neuraxial blockade. SETTINGS AND DESIGN: Prospective, cross-sectional, clinical study on consecutive patients in a tertiary care university hospital. MATERIALS AND METHODS: PCS technique, using propofol (1%) 2 ml in 2 min was used in 160 adult patients undergoing urologic procedures under central neuraxial block. We observed the time to first PCS activation by patient, duration of surgery, propofol dose, sedation score, hemodynamic stability, patient's acceptability, and the factors correlating with the PCS use. STATISTICAL ANALYSIS USED: Non parametric two-tailed Pearson's test, univariate correlation analysis for the factors favoring PCS use followed by multivariate logistic regression analysis amongst correlating factors. RESULTS: In our cohort, the majority (83%) of the patients activated PCS during surgery under central neuraxial blocks at median time of 30 min and (17%) did not activate PCS. Female patients activated sedation earlier (median 15 min) than male patients (median 30 min). All patients were hemodynamically stable and without significant side effects. Multivariate analysis showed that sedative use significantly ( P < 0.05) correlated with female gender (odds ratio-3.54 [IR-2.64 to 4.73]) and prolonged surgery (>90 min). Majority (91%) of patients rated PCS technique excellent to good. CONCLUSIONS: PCS was very well accepted by patients during central neuraxial block. Propofol regimen (2 ml in 2 min) in PCS was safe, as it caused neither apnea nor significant hypotension. PMID- 19550056 TI - Prevalence and risk factors for female sexual dysfunction in women attending a medical clinic in south India. AB - BACKGROUND: Reports from India on the prevalence and determinants of female sexual dysfunction (FSD) are scant. AIMS: To determine the prevalence and risk factors for FSD. SETTINGS AND DESIGN: A cross-sectional survey in a medical outpatient clinic of a tertiary care hospital. MATERIALS AND METHODS: We administered a Tamil version of the Female Sexual Function Index (FSFI) to 149 married women. We evaluated putative risk factors for FSD. We elicited participant's attributions for their sexual difficulties. STATISTICAL ANALYSIS: We estimated the prevalence of possible FSD and sexual difficulties from published FSFI total and domain cut-off scores. We used logistic regression to identify risk factors for possible FSD. RESULTS: FSFI total scores suggested FSD in two-thirds of the 149 women (73.2%; 95% confidence intervals [CI] 65.5% to 79.6%). FSFI domain scores suggested difficulties with desire in 77.2%; arousal in 91.3%; lubrication in 96.6%; orgasm in 86.6%, satisfaction in 81.2%, and pain in 64.4%. Age above 40 years (odds ratios [OR] 11.7; 95% CI 3.4 to 40.1) and fewer years of education (OR 1.2; 95% CI 1.0 to 1.3) were identified by logistic regression as contributory. Women attributed FSD to physical illness in participant or partner, relationship problems, and cultural taboos but none had sought professional help. CONCLUSIONS: Sexual problems suggestive of dysfunction, as suggested by FSFI total and domain scores, are highly prevalent in the clinic setting, particularly among women above 40 and those less educated, but confirmation using locally validated cut-off scores of the FSFI is needed. PMID- 19550057 TI - Renal amyloidosis secondary to childhood tuberculosis: a report of two cases. AB - Childhood renal amyloidosis is a rare entity and is mostly secondary in nature. We describe two cases of renal amyloidosis in association with childhood tuberculosis. The first case was a 10-year-old girl who presented with abdominal tuberculosis and nephrotic syndrome, while the second case was a 5-year-old boy who presented with disseminated tuberculosis and nephrotic syndrome. They were found to have amyloidosis on renal biopsy. The former was treated with anti tubercular drugs, while the latter required anti-tubercular drugs and steroids. Both the cases showed clinical improvement with remission of nephrotic syndrome. Successful treatment of tuberculosis can result in remission of nephrotic syndrome due to secondary renal amyloidosis. It is important, especially in developing countries, to be aware that tuberculosis continues to be part of the differential diagnosis of amyloidosis in children. PMID- 19550058 TI - Spontaneous closure of a traumatic intrarenal pseudoaneurysm. AB - Renal artery pseudoaneurysms caused by blunt abdominal trauma are uncommon. It requires treatment with angioembolization because of high risk of rupture. Spontaneous closure of the pseudoaneurysm is extremely unusual. We report a case of a 29-year-old male who developed a traumatic intrarenal pseudoaneurysm that later resolved spontaneously. This report provides an insight into the natural history of renal pseudoaneurysm and suggests a potential option of managing them conservatively. PMID- 19550060 TI - Declaration of Helsinki, 2008: implications for stakeholders in research. AB - The Declaration of Helsinki (DoH) was adopted by the World Medical Association (WMA) in 1964, as a statement of ethical principles, to provide guidance to physicians and other participants in medical research involving human subjects. Having undergone several amendments, the most recent version was approved on 18 October 2008, by the WMA General Assembly at Seoul, South Korea, replacing all previous versions. This version highlights issues such as, participant safety, the need to include participants from otherwise underrepresented groups, clinical trial registration, post-study access, usage of data and human tissues, compensating participants with research-related injury, and usage of placebo. In this article, we discuss the major aspects of the 2008 version, including the impact of this version on all stakeholders in research, including, investigators, ethics committee members, sponsors, authors, editors, and reviewers. PMID- 19550059 TI - Medical education in Maharashtra: the student perspective. AB - BACKGROUND: There is hardly any structured study reporting the perspective of medical students, with regard to the medical education system in Maharashtra, which is facing challenges. AIM: A perception study of students was conducted to explore the situation, challenges, and consequent solutions of medical education in Maharashtra. SETTINGS AND DESIGN: A descriptive perception study. MATERIALS AND METHODS: A structured questionnaire was e-mailed to 92 students, and interviews with seven key-informants comprising of faculty, administrators, and policy makers were conducted, to gather qualitative insights. RESULTS: Thirty seven student replies were received and analyzed. The satisfaction level of student respondents for various factors was as follows: infrastructure 18/37 (48.6%), quality of teaching 14/37 (37.8%), patient population 22/37 (59.5%), and administration 8/37 (21.6%). Ninety-two percent (34/37) of the students stated that the fundamental problem was the inability of the system to attract good, quality teachers. The reasons stated were low salaries, low level of job satisfaction, high level of bureaucracy, and high work load. CONCLUSIONS: The medical education system in Maharashtra is viewed as being stagnant. The respondents emphasized an urgent need for educational reforms, which should include better compensation for teachers, sharing of facilities between government and private medical colleges, and improved efficiency of the Medical Council of India. In the long run a public-private mix with sharing of resources may be a plausible solution. PMID- 19550061 TI - Cystic lymphoepithelial lesion of the parotid as an early indicator of HIV infection. PMID- 19550063 TI - Azathioprine induced pancytopenia: a serious complication. AB - Azathioprine is commonly used for treatment of lupus nephritis. Myelosuppression is known to occur with azathioprine, but severe pancytopenia is uncommon. A 23 year-old man with lupus nephritis was initially treated with intravenous cyclophosphamide pulses and oral prednisolone along with enalapril and frusemide. Following six months of cyclophosphamide, he was initiated on oral azathioprine as maintenance therapy. He subsequently returned with febrile neutropenia and severe bone marrow suppression. Fever responded to broad spectrum antibiotics and his counts gradually improved following granulocyte-macrophage colony stimulating factor. When last seen in October, he was symptom free and disease activity in control. We suggest that physicians remain sensitive to possibility of azathioprine induced severe bone marrow suppression. Frequent monitoring of blood counts is probably the best way to avoid this complication specially, where testing for thiopurine methyltransferase is not available. PMID- 19550062 TI - Magnetic resonance imaging findings in ophthalmoplegic migraine. PMID- 19550064 TI - Montelukast induced acute hepatocellular liver injury. AB - A 46-year-old male with uncontrolled asthma on inhaled albuterol and formoterol with budesonide was commenced on montelukast. He developed abdominal pain and jaundice 48 days after initiating montelukast therapy. His liver tests showed an increase in serum total bilirubin, conjugated bilirubin, aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase. The patient was evaluated for possible non-drug related liver injury. Montelukast was discontinued suspecting montelukast induced hepatocellular liver injury. Liver tests began to improve and returned to normal 55 days after drug cessation. Causality of this adverse drug reaction by the Council for International Organizations of Medical Sciences or Roussel Uclaf Causality Assessment Method (CIOMS or RUCAM) and Naranjo's algorithm was 'probable'. Liver tests should be monitored in patients receiving montelukast and any early signs of liver injury should be investigated with a high index of suspicion for drug induced liver injury. PMID- 19550065 TI - Targeting connexin 43 in diabetic wound healing: future perspectives. AB - The unknown mechanisms of impaired tissue repair in diabetes mellitus are making this disease a serious clinical problem for the physicians worldwide. The lacuna in the knowledge of the etiology of diabetic wounds necessitates more focused research in order to develop new targeting tools with higher efficacy for their effective management. Gap-junction proteins, connexins, have shown some promising results in the process of diabetic wound healing. Till now the role of connexins has been implicated in peripheral neuropathy, deafness, skin disorders, cataract, germ cell development and treatment of cancer. Recent findings have revealed that gap junctions play a key role in normal as well as diabetic wound healing. The purpose of this review is to provide the information related to etiology, epidemiology, clinical presentation of diabetic wounds and to analyze the role of connexin 43 (Cx43) in the diabetic wound healing process. The current control strategies and the future research challenges have also been discussed briefly in this review. PMID- 19550067 TI - Need to regulate burgeoning clinical research courses. PMID- 19550066 TI - Cut-off value for PSA: do we need a change? PMID- 19550068 TI - The value of communication. PMID- 19550069 TI - Mentoring medical student research through studentships and fellowships: reflections from India. PMID- 19550070 TI - Comments on neurolinguistic study. PMID- 19550071 TI - Rifampicin-induced severe headache in HIV-tuberculosis coinfected patient. PMID- 19550072 TI - Recurrent vomiting and anemia: an unusual clinical condition caused by midge larva. PMID- 19550073 TI - Expression and function of IA-2 family proteins, unique neuroendocrine-specific protein-tyrosine phosphatases. AB - IA-2 (also known as islet cell antigen ICA-512) and IA-2 beta (also known as phogrin, phosphatase homologue in granules of insulinoma) are major autoantigens in insulin-dependent diabetes mellitus (IDDM). Autoantibodies against both proteins are expressed years before clinical onset, and they become predictive markers for high-risk subjects. However, the role of these genes in the IDDM pathogenesis has been reported fairly negative by recent studies. IA-2 and IA-2 beta are type I transmembrane proteins that possess one inactive protein-tyrosine phosphatase (PTP) domain in the cytoplasmic region, and act as one of the constituents of regulated secretory pathways in various neuroendocrine cell types including pancreatic beta-cells. Existence of IA-2 homologues in different species suggests a fundamental role in neuroendocrine function. Studies of knockout animals have shown their involvement in maintaining hormone content, however, their specific steps in the secretory pathway IA-2 functions as well as their molecular mechanisms in the hormone content regulation are still unknown. More recent studies have suggested a novel function showing that they contribute to pancreatic beta-cell growth. This review attempts to show the possible biological functions of IA-2 family, focusing on their expression and localization in the neuroendocrine cells. PMID- 19550074 TI - Unfavorable blood rheology is closely associated with arterial stiffness in obese patients. AB - Little information on the relationship between blood rheology and atherosclerosis indicators in obese patients is available. We examined blood rheology as assessed by the blood passage time (BPT) with the microchannel method in 109 obese patients. BPT was correlated well with the extent of each metabolic syndrome component. A multivariate regression analysis revealed that the independent contributors to BPT were pulse-wave velocity, an index of arterial stiffness, body mass index and red blood cell. Furthermore, weight reduction intervention significantly decreased BPT. Assessment of rheology may be associated with pulse wave velocity, and useful to manage obese patients. PMID- 19550075 TI - Administration of conophylline and betacellulin-delta4 increases the beta-cell mass in neonatal streptozotocin-treated rats. AB - The present study was conducted to examine the effect of administration of conophylline (CnP) and betacellulindelta4 (BTCdelta4) on the beta-cell mass in neonatal streptozotocin-treated rats (neonatal STZ rats). STZ (100 microg/g) was injected into neonatal rats, and then CnP (2 microg/g) and/or BTCdelta4 (200 pmol/g) were administered to neonatal STZ rats for 1 week. The plasma glucose concentration was monitored, and an intraperitoneal glucose tolerance test (ipGTT) was performed on day 8 and at 8 weeks after the STZ injection. In neonatal STZ rats treated with control solution (S group), the plasma glucose concentration increased for several days after the STZ injection, returned to nearly normal levels, and then increased gradually after six weeks of age. Eight weeks after the STZ-injection, the plasma glucose concentration was increased significantly compared to that of normal rats. The glucose response to ipGTT was significantly reduced in neonatal STZ rats treated with CnP (CnP group), BTCdelta4 (delta4 group) and CnP+BTCdelta4 (CnP+delta4 group). The beta-cell mass and the insulin content of the pancreas were significantly increased in the CnP group and delta4 group. The effect of CnP+delta4 was greater than that of CnP alone or BTCdelta4 alone. CnP+BTCdelta4 significantly increased the number of PDX 1-positive ductal cells and the number of insulin/BrdU double-positive ductal cells. These results indicate the efficacy of CnP and BTCdelta4 in increasing the beta-cells mass of neonatal STZ-treated rats. PMID- 19550076 TI - Single nucleotide polymorphism analysis of the follicle-stimulating hormone (FSH) receptor in Japanese with male infertility: identification of codon combination with heterozygous variations of the two discrete FSH receptor gene. AB - AIMS: Dysfunction of the FSH receptor (FSHR) may be involved in some form of male infertility with azoospermia or oligozoospermia. We assessed the discrete codon combination with homo/heterozygous variation of the exon 10 in the FSHR gene. METHODS: The genotype of codon 307 and codon 680 were analysed in 352 patients with idiopathic male infertility and 145 men with proven fertility. RESULTS AND CONCLUSION: There was no significant difference in the distributions of each homozygous codon 307 or 680 between these two groups as reported in the literature. However, the population with heterozygous combinations Thr/Ala (codon 307) and Ser/ Asn (codon 680) comprised 26% (38/146) and 44.9% (157/343) in subjects with proven fertility and idiopathic infertile men, respectively. Moreover, the heterozygous genotype Thr/Ala-Ser/Asn was significantly increased in infertile patients compared with the controls. This finding showed that the combination of heterozygous FSHR can be responsible for male infertility. PMID- 19550077 TI - Vitamin K2 suppresses proliferation and motility of hepatocellular carcinoma cells by activating steroid and xenobiotic receptor. AB - Vitamin K2, known as a cofactor for gamma-carboxylase, also serves as a ligand of a nuclear receptor, Steroid and Xenobiotic Receptor (SXR). Several clinical trials revealed that vitamin K2 reduced de novo formation and recurrence of hepatocellular carcinoma (HCC). To examine the role of SXR in HCC as a receptor activated by vitamin K2, the cells stably overexpressing SXR were established using a HCC cell line, HuH7. Overexpression of SXR resulted in reduced proliferation and motility of the cells. Further suppression of proliferation and motility was observed when SXR overexpressing clones were treated with vitamin K2. These results suggest that the activation of SXR could contribute to tumor suppressive effects of vitamin K2 on HCC cells. PMID- 19550078 TI - Prevalence of TSH receptor and Gsalpha mutations in 45 autonomously functioning thyroid nodules in Japan. AB - Somatic mutations of the thyrotropin receptor (TSHR) gene and the gene encoding the alpha subunit of the stimulatory GTP-binding protein (Gsalpha) are the main cause for autonomously functioning thyroid nodules (AFTN) in iodine-deficient regions of the world. In iodine-sufficient regions, including Japan, the genetic relevance of AFTN is unclear. In a series of 45 Japanese subjects with AFTN, exons 9 and 10 of the TSHR and exons 7-10 of Gsalpha , where the activating mutations have been found, were analyzed using direct sequencing. We found 29 somatic mutations: 22 in the TSHR gene and 7 in the Gsalpha gene. The most frequent mutation in TSHR was Met453Thr (10 cases), followed by clustered residues from codons 630 through 633 on TSHR (7 cases). Mutations of Gsalpha were detected at codon 201 in 5 cases and at codon 227 in 2 cases. No patients had coexistent TSHR and Gsalpha mutations in the same nodule. All mutated residues but one, which was deleted at codon 403 on the TSHR gene, are constitutively active. The prevalences of a germline polymorphism of Asp727Glu on the TSHR gene and incidental papillary thyroid carcinoma in thyroid surgical specimens were similar to those reported in other studies. In the present study, more than half of the cases with AFTN had a somatic activating mutation either of the TSHR or Gsalpha gene, despite their high iodine intake. PMID- 19550079 TI - Plasma adiponectin levels are increased despite insulin resistance in corticotropin-releasing hormone transgenic mice, an animal model of Cushing syndrome. AB - Adiponectin (AdN), an adipokine derived from the adipose tissue, has an insulin sensitizing effect, and plasma AdN is shown to be decreased in obesity and/or insulin resistant state. To clarify whether changes in AdN are also responsible for the development of glucocorticoid-induced insulin resistance, we examined AdN concentration in plasma and AdN expression in the adipose tissue, using corticotropin-releasing hormone (CRH) transgenic mouse (CRH-Tg), an animal model of Cushing syndrome. We found, unexpectedly, that plasma AdN levels in CRHTg were significantly higher than those in wild-type littermates (wild-type: 19.7+/-2.5, CRH-Tg: 32.4+/-3.1 microg/mL, p<0.01). On the other hand, AdN mRNA and protein levels were significantly decreased in the adipose tissue of CRH-Tg. Bilateral adrenalectomy in CRH-Tg eliminated both their Cushing's phenotype and their increase in plasma AdN levels (wild-type/sham: 9.4+/-0.5, CRH-Tg/sham: 15.7+/ 2.0, CRH-Tg/ADX: 8.5+/-0.4 microg/mL). These results strongly suggest that AdN is not a major factor responsible for the development of insulin resistance in Cushing syndrome. Our data also suggest that glucocorticoid increases plasma AdN levels but decreases AdN expression in adipocytes, the latter being explained possibly by the decrease in AdN metabolism in the Cushing state. PMID- 19550080 TI - Predominant expression of mutated allele of the succunate dehydrogenase D (SDHD) gene in the SDHD-related paragangliomas. AB - Recent studies indicate that succinate dehydrogenase (SDH) genes B, C, or D are, at least partly, involved in the pathogenesis of pheochromocytoma or paraganglioma. Of these three genes, the SDHD gene mutation is most closely related with paragangliomas of the neck. Here we describe a case of an SDHD related paraganglioma, in which we studied the molecular characteristics of an SDHD mutation to evaluate the involvement of SDHD in neck paragangliomas. Genetic testing revealed a heterozygous G106D mutation in the SDHD gene. In the tumor tissue, loss of heterozygosity was demonstrated by real time polymerase chain reaction (PCR). In the present case of SDHD mutated paragangliomas, wild type SDHD gene expression was markedly reduced possibly due to loss of heterozygosity not due to imprinting of SDHD gene in the tumors. PMID- 19550081 TI - Effect of alumina air-abrasion on mechanical bonding between an acrylic resin and casting alloys. AB - This study examined the effect of alumina air-abrasion with different pressure on bonding between an acrylic resin and casting alloys. Disk specimens (8 and 10 mm in diameter) were cast from a silver-palladium-copper-gold (Ag-Pd-Cu-Au, Castwell M.C.12) alloy and a titanium-aluminum-niobium alloy (Ti-6Al-7Nb, T-Alloy Tough). The disks were air-abraded with alumina particles (50-70 microm) under different air-pressures (0 unabraded, 0.1, and 0.6 MPa). The disk pairs were bonded together with a tri-n-butylborane (TBB)-initiated acrylic resin, and shear bond strengths were determined both before and after thermocycling. Bond strength varied from a maximum of 37.1 MPa to a minimum of 3.6 MPa for the Ag-Pd-Cu-Au alloy, whereas bond strength to Ti-6Al-7Nb alloy ranged from 34.7 MPa to 0.1 MPa. Specimens abraded with 0.6 MPa pressure recorded the greatest post-thermocycling bond strength (21.7 MPa and 17.9 MPa), and unabraded specimens showed the lowest strength (3.6 MPa and 0.1 MPa) for both alloys. Post-thermocycling bond strength to the Ag-Pd-Cu-Au alloy was higher than that to the Ti-6Al-7Nb alloy under identical air-abrading conditions. It can be concluded that alumina air-abrasion with an air-pressure of 0.6 MPa is effective in enhancing retentive characteristics of the TBB-initiated resin joined to the alloys. PMID- 19550082 TI - Facilitation of reflex swallowing from the pharynx and larynx. AB - To evaluate the cooperative effect of afferent signals from the pharynx and larynx on reflex swallowing, the interactive effect of afferent signals from the pharyngeal branch of the glossopharyngeal nerve (GPN-ph) and superior laryngeal nerve (SLN) was analyzed in detail in urethane-anesthetized rats. The electromyographic activity of the mylohyoid muscle was recorded as an indicator of swallowing activity. The onset latency of reflex swallowing was measured to evaluate the effects of electrical stimulation of these nerves, and found to become shorter following an increase in the GPN-ph and/or SLN stimulus frequency. During simultaneous electrical stimulation of the GPN-ph and SLN (frequency: 5-10 Hz, intensity: 30 muA, duration: 1.0 ms for each), the onset latency of reflex swallowing became shorter than that for stimulation of each nerve independently. The present findings suggest that spatiotemporal summation of afferent signals from the GPN-ph and SLN results in an increase of motoneuronal activity in the medullary swallowing center, thus enhancing reflex swallowing. PMID- 19550083 TI - Involvement of phosphodiesterase 4 in beta-adrenoceptor agonist-induced amylase release in parotid acinar cells. AB - beta-Adrenoceptor activation increases intracellular cAMP levels and consequently induces exocytotic amylase release in parotid acinar cells. Phosphodiesterase (PDE) catalyses the hydrolysis of cAMP, which terminates the downstream signaling of this second messenger. We investigated the involvement of PDE4, a cAMP-PDE, in beta-adrenoceptor agonist-induced amylase release in mouse, rat and rabbit parotid acinar cells by using the specific PDE4 inhibitor rolipram. cAMP-PDE activity was detected in mouse, rat and rabbit parotid acinar cells. In the presence of rolipram, cAMP-PDE activity was reduced by about 31%, 38% and 33% in mouse, rat and rabbit parotid acinar cells, respectively. The increase in cAMP levels induced by the beta-adrenoceptor agonist isoproterenol was enhanced in the presence of rolipram in mouse, rat and rabbit parotid acinar cells. Isoproterenol induced amylase release, but not constitutive amylase release, was also enhanced in the presence of rolipram in mouse, rat and rabbit parotid acinar cells. These results suggest that the rolipram-sensitive cAMP-PDE, PDE4, is involved in beta adrenoceptor agonist-induced amylase release in parotid acinar cells. PMID- 19550084 TI - Comparison between gutta-percha and Resilon retreatment. AB - Retreatment procedures in endodontic practice require complete removal of the original root filling materials. The aim of this study was to compare the amount of residual filling material on the walls of root canals that were obturated with gutta-percha or Resilon. Thirty extracted single-rooted human teeth were selected and instrumented by RaCe rotary instruments up to MAF #35. They were randomly divided into two groups of 15 teeth each. Group 1 was obturated with gutta-percha and AH26 sealer and group 2 was obturated with the Resilon/Epiphany system by lateral condensation technique. All canals were then retreated using Gates Glidden drills, MAF #40 and chloroform. The teeth were split longitudinally and photographs were taken by a digital camera connected to a stereomicroscope and then samples were analyzed. The amount of residual material and time of retreatment were determined in each group at three levels in the canal. Data were analyzed by Mann-Whitney and Friedman tests. The Resilon group had significantly more residual material in the canal than the Gutta-percha group (P = 0.009). According to the level of the canal, the differences were statistically significant (P = 0.013) only in the Resilon group. Time required for retreatment in the two groups was not significantly different (P = 0.381). Orthograde retreatment of Resilon was less efficient than that of gutta-percha. PMID- 19550085 TI - Effect of phosphate buffer saline on coronal leakage of mineral trioxide aggregate. AB - This study was carried out to compare the bacterial leakage of MTA used as a root end filling material when it was kept in phosphate buffer saline (PBS) or normal saline. In this study, 72 freshly extracted teeth were used. The roots were randomly divided into four experimental groups of 15 each (groups I and II gutta percha obturation + MTA, groups III and IV only MTA) and two positive and negative control groups of six each. The samples in groups I and III were kept in normal saline for 1 month while the samples in groups II and IV were kept in PBS. Enterococcus faecalis was used for determination of the bacterial penetration. Kaplan-Meier survival curve and chi(2) test were employed for data analysis. The obturated samples with root-end filling showed significantly longer duration of resistance to bacterial penetration than canals without obturation (P < 0.05). The roots that were placed in PBS (groups II and IV) showed significantly less bacterial penetration in comparison with the roots that were stored in normal saline (P < 0.05). In conclusion, MTA, which acts as a bioactive material, should be placed in a synthetic tissue fluid before any leakage evaluation. PMID- 19550086 TI - Morphological characteristics of microcirculation in oral lichen planus involving the lateral border of the tongue. AB - Oral lichen planus (OLP) is a chronic mucosal condition commonly encountered in dental practice. Lichen planus is believed to represent an abnormal immune response in which epithelial cells are recognized as foreign, secondary to changes in the antigenicity of the cell surface. It has various oral manifestations. The aim of the study was to evaluate the morphological characteristics of microcirculation of lingual lichen planus (LLP). Twenty patients (10 patients with LLP and 10 healthy patients) were examined by means of videocapillaroscopy. The left margin of the lingual mucosa was examined in each patient. The capillary loop length, loop diameter, and capillary density were analyzed on every capillaroscopic image. The results obtained using videocapillaroscopic software were subjected to statistical analysis. The density, loop length, and total diameter showed statistically significant differences. A remarkable increase in capillary density was shown. The microcirculation of LLP can be studied "in vivo" with the help of videocapillaroscopy, and it could be an indicator of the evolutionary condition of LLP, useful for the evaluation of the evolution or regression phases of the disease. PMID- 19550087 TI - Impact of different periodontitis case definitions on periodontal research. AB - Different periodontitis definitions have been used in periodontal research. This study assessed the impact of case definition on the prevalence and extent rates of periodontitis. A data set including 340 periodontal records, collected in Belo Horizonte, Brazil, was used. Periodontitis was defined as: 1) one site with probing depth (PD) >or= 4 mm; 2) clinical attachment level (CAL) >or= 5 mm in >or= 4 sites + one site with PD >or= 4 mm; 3) CAL >or= 6 mm in >or= 2 teeth + one site with PD >or= 5 mm; 4) >or= 4 teeth with >or= 1 sites with PD >or= 4 mm + CAL >or= 3 mm; 5a) interproximal CAL or PD >or= 4 mm at >or= 2 sites, not on the same tooth; and 5b) interproximal CAL of >or= 6 mm at >or= 2 sites, not on the same tooth + PD >or= 5 mm at >or= 1 proximal site. Definition 5 was determined to be the gold standard and the definitions were compared by means of agreement, sensitivity, specificity, and positive and negative predictive values. Prevalence and extent rates greatly varied, from 13.8% to 65.3% and from 9.7% to 55.6%, respectively. The use of different case definitions has a great impact on the prevalence and extent rates of periodontitis. PMID- 19550088 TI - Evaluation of microleakage following application of a dentin bonding agent as root canal sealer in the presence or absence of smear layer. AB - The aim of this study was to compare the apical leakage of roots obturated with gutta-percha using either an epoxy resin sealer (AH26) or a dual cure dentin binding agent (Excite DSC) as sealer in the presence or absence of smear layer with fluid filtration method. The canals of eighty-six, single-rooted premolars were instrumented until a #40 K-file fit at working length and then randomly divided into four groups (n = 20) with the remaining six used as controls. Groups 1 and 2 were filled with gutta-percha using AH26 as sealer; groups 3 and 4 were filled with gutta-percha and Excite DSC as sealer. Groups 1 and 3 were smear layer-positive, while group 2 and 4 were designated as smear layer-negative. After 3 days and 3 months, the samples were connected to a fluid filtration system. Analysis of data with the paired t-test showed that microleakage in AH26 groups (with and without smear layer) decreased significantly at 3 months compared to 3 days; however, in the DBA groups, the amount of microleakage at 3 days and 3 months was not significantly different. According to the results of this study, DBA (Excite DSC) had better apical sealing ability and could be applied clinically. PMID- 19550089 TI - Surface properties of an indirect composite polymerized with five laboratory light polymerization systems. AB - A study was conducted to evaluate the influence of laboratory light polymerization systems on the post-curing properties of a composite. An indirect composite (Sinfony) was polymerized with five polymerization systems (Visio system, Hyper LII, Pearlcure Light, Twinkle MIII, and UniXS II) using nine polymerization modes. After light exposure, Knoop hardness number, wear depth, and changes in gloss were determined. The highest hardness number was recorded with the use of the Hyper LII (120 s) and Pearlcure Light (120 s) units, whereas the lowest value was obtained with the Visio system and UniXS II (60 s). Six groups demonstrated comparable as well as higher wear resistance to toothbrush abrasion (Hyper LII 60 and 120 s, UniXS II 120 s, Pearlcure Light 60 and 120 s, and Twinkle MIII 120 s), and two groups exhibited lower wear resistance (Visio system and UniXS II 60 s). Gloss of the composite was not dependent on the polymerization mode used before wear testing. However, surface gloss was significantly reduced by toothbrush dentifrice abrasion. Within the limitations of the present experiment, it can be concluded that the Sinfony composite can be polymerized sufficiently with high-intensity light polymerization units. PMID- 19550090 TI - Radiopacity of conventional, resin-modified glass ionomer, and resin-based luting materials. AB - The purpose of the present study was to evaluate the radiopacity of currently available dental luting materials. Five conventional cements, six resin-modified glass ionomers (RMGIs), two methyl methacrylate (MMA)-based acrylic resins (eight shades), and nine composite luting materials were evaluated. Radiographs of the specimens were taken together with tooth slices and aluminum step wedges. The density of the specimens was determined with a densitometer and was expressed in terms of the equivalent thickness of aluminum per 2.0-mm unit thickness of specimen. The radiopacity values for human enamel and dentin were 4.3 and 2.3 mm Al/2.0 mm specimen, respectively. The values for materials ranged from 5.1 to 12.9 for conventional luting materials, from 3.4 to 6.3 for RMGIs, from less than 0.5 to 7.3 for MMA resins, and from 2.3 to 9.9 for the composite luting materials. A zinc phosphate cement showed the highest value (12.9), whereas five shades of MMA resin resulted in the lowest value (less than 0.5). Two RMGIs and three composite luting materials exhibited radiopacity values between those of enamel (4.3) and dentin (2.3). It can be concluded that the radiopacity value of luting materials varies considerably, and that care must be taken when selecting luting materials, considering the material composition of restorations. PMID- 19550091 TI - A radiographic study on the visualization of the anterior loop in dentate subjects of different age groups. AB - The anterior loop is defined as where the mental neurovascular bundle crosses anterior to the mental foramen then doubles back to exit the mental foramen. It cannot be seen clinically but can be detected in 11-60% of panoramic radiographs. As this anatomical structure is important in determining the placement position of endosseous implants in the mandibular premolar region, a pilot study was undertaken to determine its visibility on dental panoramic radiographs in dentate subjects of various age groups. One or more anterior loops were visible in 39 (40.2%) radiographs encompassing 66 sites (34.4%). Interestingly, anterior loops were most commonly observed bilaterally, followed by on the right side of the mandible only. An anterior loop on the left side only was observed in just 1 radiograph. Visibility of anterior loops reduced as the age of subjects increased. More than half (58.1%) of subjects aged 20-29 years exhibited at least one anterior loop; this gradually reduced to only 15 percent of subjects aged 50 and older. There was no association between visualization of the anterior loop and subject gender. PMID- 19550092 TI - Site-specificity of pain sensitivity to intraoral anesthetic injections in children. AB - A total of 455 children (242 males and 213 females) aged 5-6 years were investigated for their pain reactions to maxillary and mandibular infiltration anesthesia, middle superior alveolar nerve block, posterior superior alveolar nerve block, greater palatine nerve block, nasopalatine nerve block, and inferior alveolar nerve block, and the responses were quantified using the sound, eye and motor (SEM) scale. Administration of nasopalatine nerve block produced maximum pain (median SEM score: 10) while that of posterior superior alveolar nerve block and inferior alveolar nerve block was accompanied by minimum pain (3 and 4, respectively). There was no significant gender-specific difference in pain reactions (P = 0.39). Administration of local anesthesia in the maxilla was more painful than injections into the mandible (7 versus 5, P < 0.05). Furthermore, infiltration into the anterior and posterior segments of the maxilla produced maximum and minimum pain reactions, respectively (8 versus 3, P < 0.001). It seems that the anatomical location of an injection is one of the most important determinants of pediatric pain reaction, and that mandibular injections are generally less painful. Prioritization of treatment by consideration of site dependent variability in pain sensitivity may help to achieve optimal behavioral control during dental treatment in young children. PMID- 19550093 TI - Does dental anxiety influence oral health-related quality of life? Observations from a cross-sectional study among adults in Udaipur district, India. AB - We investigated the effect of dental anxiety and dental visiting habits, as well as various socio-demographic variables, on oral health-related quality of life (OHQoL) among subjects aged 15-54 years living in Udaipur district, India. The total sample size was 1235 individuals and a stratified cluster sampling procedure was employed to collect the representative sample. Dental anxiety and oral health-related quality of life were assessed using the Corah Dental anxiety scale and the OHQoL-UK(W) questionnaire, respectively. The majority of the female and older individuals showed higher dental anxiety than their male and younger counterparts. Stepwise linear regression analysis revealed that the best predictors of dental anxiety were, in descending order, occupation, gender and education, which provided a variance of 10.3%. Females were more likely to have poor OHQoL than males. Dental anxiety had a significant influence on OHQoL, people with high dental anxiety being 2.34 times more likely to present poor OHQoL than those having low anxiety. Furthermore, it was found that those who never visited a dentist had an odds ratio of 1.62 for poor OHQoL relative to those who had visited a dentist within the last 12 months. Dental anxiety differed significantly with age and dental visiting practices, and had a significant impact on oral health-related quality of life after controlling for other variables. PMID- 19550094 TI - Panavia F: the role of the primer. AB - This study evaluated the rate of polymerization (R(p)) and degree of conversion (DC) of Panavia F when self- or dual-activated, and the influence of either using or not using a primer containing co-initiators (ED Primer) mixed with the material. The conversion reaction was monitored using real-time infrared spectroscopy with an attenuated total reflectance device. The cement was mixed, put onto the device and irradiated for 20 s (dual-cured). A self-cured group was also prepared. Similarly, dual- and self-cured groups were evaluated after mixing ED Primer with the cement. The DC was monitored for 1 h and the R(p) was calculated. For the dual-cured mode, the most significant effect of ED Primer was to reduce the R(p)(max). For the self-cured mode, the primer was shown to be essential for polymerization of Panavia F: without ED Primer, the reaction started only after approximately 500 s, with a final DC of 50%, whereas a continuous increase in conversion was observed for the group mixed with ED Primer, with a final DC of 74%. The ED primer is essential for proper polymerization of Panavia F; the cement becomes independent of the light when the primer is used. PMID- 19550095 TI - Exploring the role of Th1 cytokines: interleukin-17 and interleukin-18 in periodontal health and disease. AB - There are conflicting data regarding the role of interleukin (IL)-17 in periodontal health and disease. However, IL-18 levels are known to increase with the severity of periodontal disease. The present study was performed to explore the role of these proinflammatory cytokines in periodontal disease progression, and also to clarify the effect of periodontal treatment on their concentration. Sixty age- and gender-matched subjects were divided into three groups each consisting of 20 subjects on the basis of gingival index (GI), probing pocket depth (PPD), clinical attachment loss (CAL) and radiological parameters (bone loss): healthy (group 1), gingivitis (group 2) and chronic periodontitis (group 3), while group 3 patients after treatment constituted group 4. GCF samples collected from all the groups were assayed by enzyme-linked immunosorbent assay to estimate the levels of IL-17 and IL-18. IL-18 levels in GCF increased proportionally with the severity of periodontal disease, and decreased after treatment. However, IL-17 levels in GCF were nearly zero. Since our data indicate an absence of IL-17 in GCF, it cannot be considered as a biomarker of periodontal disease progression, at least in Indian populations. However, IL-18 appears to be a good inflammatory biomarker. PMID- 19550096 TI - Color stability of indirect composite materials polymerized with different polymerization systems. AB - The purpose of the current study was to evaluate the color stability of two indirect composite materials (Sinfony and Pearleste) polymerized with different laboratory polymerization systems. Disk specimens were prepared with their proprietary polymerization systems (Visio and Pearlcure systems) or with a metal halide light polymerization unit (Hyper LII) for 60, 120, and 180 s. After storage at 37 degrees C for 24 h, the specimens were immersed in either purified water or tea. Color change between baseline evaluation and after 4 weeks was determined with a dental chroma meter (ShadeEye NCC) using black and white backgrounds. CIE 1976 L(*)a(*)b(*) values were determined, and they were converted into DeltaE(*)(ab) values. The DeltaE(*)(ab) value of the Sinfony material immersed in tea was the highest when the material was polymerized with the proprietary Visio system. The Pearleste material immersed in purified water and tea was not affected substantially by the polymerization systems. Among the 12 groups polymerized with the Hyper LII units, DeltaE(*)(ab) values of 11 groups were significantly lower for the Pearleste material than for the Sinfony material. It can be concluded that the Pearleste material was stable against color change when the material was polymerized with either the Pearlcure system or with the Hyper LII unit. PMID- 19550097 TI - Comparison of periodontal health status and oral health behavior between Japanese and Chinese dental students. AB - A survey was carried out to compare periodontal health status and oral health behavior between Japanese and Chinese dental students. Subjects consisted of 118 students at Nihon University School of Dentistry and 92 students at the school of Stomatology, Nanjing Medical University. Saliva occult blood test was performed to classify whether subjects may have periodontal disease. Further questionnaires were given to evaluate different lifestyles and oral hygiene habit. The positive rate of the saliva occult blood test in Japanese dental students was 13.6%, and that of Chinese dental students was 43.5%. Bleeding from gingiva as a subjective symptom was as follows: Japansese 7.6%, Chinese 37.0%. Japanese dental students brushed for 13.5 min each day. The rate for Chinese students was 4.6 min. Use of interdental devices was as follows: Japanese 33.1%, Chinese 7.6%. Differences of periodontal disease rates between Japanese and Chinese dental students are thought to be differences in oral hygiene, indicating the need for improvements in hygiene measures in Nanjing City. The establishment and strengthening of oral hygiene education, including the importance of tooth brushing for prevention of periodontal disease, has been proposed. PMID- 19550098 TI - The significance of podoplanin expression in human inflamed gingiva. AB - Podoplanin, a transmembrane glycoprotein, has been considered to be expressed specifically by lymphatic endothelial cells. However, recent studies have shown that the protein is expressed in a variety of normal as well as neoplastic tissues, and that its expression might be related to cell migration and invasion. In this study, we examined podoplanin expression in inflamed gingival tissues using an immunohistochemical method. Positive immunoreactivity for podoplanin was found in the cell membrane and cytoplasm of basal cells of oral gingival epithelium when severe inflammatory cell infiltration was present in the connective tissue just under the epithelium. When inflammatory changes were weak or absent, little or no reactivity for podoplanin in the basal cells was observed. Positive reactivity for podoplanin was also detected in basal cell extensions. Surprisingly, strong immunoreactivity for podoplanin was observed in all layers of oral sulcular and junctional epithelia associated with severe inflammatory reaction in the connective tissue. These findings suggest that increased expression of podoplanin in gingival epithelium is related to the progression of chronic periodontitis. PMID- 19550100 TI - Treatment of large ameloblastic fibroma: a case report. AB - Ameloblastic fibroma (AF) is an extremely rare true mixed benign tumor that can occur in either the mandible or the maxilla, but is most frequently found in the posterior region of the mandible. It usually occurs in the first two decades of life and is associated with tooth enclosure, causing a delay in eruption or altering the dental eruption sequence. AF is diagnosed on routine radiographic evaluation and is clinically and radiographically similar to ameloblastic fibrodontoma and odontoma, which makes an accurate diagnosis mandatory. There is controversy in the literature as to whether treatment should be conservative or agressive. A conservative treatment strategy, such as enucleation and curettage, is usually sufficient. However, extensive lesions require radical treatment. We describe a case of ameloblastic fibroma with a very unusual clinical manifestation: it demonstrated considerable extension but no associated impacted tooth, was located in the anterior region of the mandible, and became symptomatic in the fifth decade of life. A radical surgical approach was taken, with immediate reconstruction. PMID- 19550099 TI - Induction of micronuclei in buccal mucosa on chewing a mixture of betel leaf, areca nut and tobacco. AB - Betel quid containing areca nut and chewing tobacco is used in many parts of India. In this study we evaluated the micronuclei (MN) in buccal mucosa of healthy individuals from southern India, who were regularly chewing a mixture of betel leaf, areca nut and tobacco. A total of 44 subjects were examined. The study population included 15 chewers, 14 chewers with smoking habit and 15 controls with the mean age of 38.57 +/- 0.54, 34.50 +/- 0.95, and 33.28 +/- 0.89 years, respectively. The mean percentage of MN was 1.90 +/- 1.03 in chewers, 2.00 +/-1.12 in chewers with smoking habits and 0.81 +/- 0.66 in controls. There was no significant difference between the mean percentages of the two experimental groups. It can be concluded that a mixture of betel leaf, areca nut, and tobacco is unsafe for oral health. PMID- 19550102 TI - Noonan syndrome: a case report. AB - Cranio-dento-facial findings in a case of Noonan syndrome in a 13-year, 3-month old male are reported. Oral examination of the patient revealed a narrow, high arched palate, malocclusion and congenitally missing teeth. Even though cephalometric measurements showed an increased vertical facial pattern (GoGn/SN = 38.46 degrees ), a severe deep bite (9 mm) was evident. Noonan syndrome is characterized by ocular, facial, cardiac, and dental anomalies, although it is often unrecognized by clinicians. Although differential diagnosis of this syndrome is difficult, the cranio-dento-facial structures show specific findings that can be diagnosed by a dental specialist. The present report describes a new case of this syndrome from a dental viewpoint. PMID- 19550101 TI - Occurrence of talon cusp on a geminated maxillary central incisor: a case report. AB - This article reports an uncommon case of talon cusp on a geminated permanent maxillary central incisor, including details of the clinical and radiographic findings and treatment. A 28-year-old woman presented at the university dental clinic, and intraoral examination revealed a maxillary central incisor of abnormal size with a prominent accessory cusp on the lingual aspect. The case was diagnosed as talon cusp on a geminated tooth. Early diagnosis of this anomaly is important since it may cause clinical problems such as esthetic concerns, caries and tooth crowding. In the present case, these anomalies did not complicate the subsequent endodontic treatment. PMID- 19550103 TI - Intraosseous myofibroma simulating an odontogenic lesion. AB - Myofibroma is a rare benign nodular tumor of the soft tissues, bones, or internal organs, and may affect both children and adults. It is mostly found in the head and neck region, although uncommon in the jaw bones, where only a few lesions have been described. Radiographically, it may appear as a well-defined unilocular radiolucent entity simulating a cystic or odontogenic lesion. Histopathologic findings alone may be insufficient for a final diagnosis, and immunostaining for markers such as vimentin, alpha-smooth muscle actin, desmin and S-100 may be required. Intraosseous myofibroma is a diagnostic challenge and requires careful histopathologic, immunohistochemical and radiographic correlation. The present article describes a conservatively treated intraosseous myofibroma that occurred in a 7-year-old girl. The clinical, radiographic, histopathologic and immunohistochemical features are discussed. PMID- 19550104 TI - Chondrolipoma of the tongue: a case report. AB - Chondrolipomas are benign mesenchymal tumors characterized by the proliferation of mature adipocytes associated with variable amounts of mature cartilaginous tissue. We describe here a case of chondrolipoma of the tongue in a 30-year-old man. The patient reported the presence of the lesion for approximately 10 years. Intraoral clinical examination revealed a nodular, sessile, pink mass of firm consistency, which was located along the midline of the posterior region on the dorsal surface of the tongue. An excisional biopsy was performed and histopathological examination revealed a well-delimited proliferation of mature adipocytes arranged in lobules and separated by fibrous connective tissue septa. Islands of mature cartilaginous tissue were identified in close proximity to the fibrous connective tissue septa, as well as lacunae filled with chondrocytes. No myxoid areas, lipoblasts, atypias, hyperchromasia or mitosis were detected. After 5 years of follow-up, the patient shows no signs of recurrence. Chondrolipomas are uncommon in the oral cavity, with only nine cases being reported in the international literature (Pubmed Database). PMID- 19550105 TI - Treatment of multiple gingival recessions using subepithelial connective tissue grafting with a single-incision technique. AB - The increasing interest in esthetics and the subsequent need to solve related problems such as dentin hypersensitivity and root caries have favored the development of many surgical techniques that permit the coverage of exposed roots. In this case, a 47-year-old female patient presented seeking resolution of dentin hypersensitivity with multiple gingival recessions in the upper anterior region. A coronally positioned flap combined with connective tissue grafting using a single-incision technique was applied to achieve root coverage. The treated site showed improvement of root coverage with reduction of dentin hypersensitivity without any probing defect or significant complication. The single-incision technique may be an option for harvesting of graft material in the treatment of multiple gingival recessions. PMID- 19550106 TI - Molecular cloning of paired related homeobox 2 (prx2) as a novel pituitary transcription factor. AB - This study aimed to identify protein(s) that bind(s) to the highly AT-rich sequence of porcine Fshb promoter region -852/-746 (named Fd2) by the Yeast One Hybrid Cloning System and finally a paired related homeodomain transcription factor, Prx2, known as a key factor for skeletogenesis was cloned. RT-PCR analysis of fetal and postnatal porcine pituitaries demonstrated that Prx2 starts to be expressed at around fetal days 40-50 just before the beginning of Lhb expression and that the level of Prx2 increases after birth. Immunohistochemical analysis of the prepubertal porcine pituitary revealed that some Prx2-positive cells overlap some Lh beta-positive cells. Transient transfection assay using non pituitary CHO cells and pituitary tumor-derived LbetaT2 cells revealed that Prx2 plays a cell-type dependent role in modulation of the Fshb promoter, showing stimulation in CHO cells and repression in LbetaT2 cells via the regions of Fd2 and -596/-239. The binding ability of Prx2 to the regions of Fd2 and -596/-239 was confirmed by electrophoretic mobility shift assay. DNase I footprinting revealed that broad regions of Fd2 were bound by Prx2 and that -596/-239 contained seven Prx2-binding sites. The SELEX method using a random N15-mer oligonucleotide pool demonstrated that Prx2 monomer binds to a TAATT motif, which is present in Fd2 and -596/-239. However, the binding of Prx2 to TAATT with a single molecule and its inverted repeat with two molecules could not induce transcriptional activation, indicating that the Prx2-dependent transcriptional modulation demonstrated in cultured cells is not introduced by Prx2 alone. Thus, this study demonstrated for the first time that Prx2 is expressed in the pituitary gland and at least in a part of gonadotropes in which Prx2 may play a role in repression of the Fshb gene. PMID- 19550107 TI - Alteration in anxiety with relation to the volume of the locus ceruleus in progranulin-deficient mice. AB - The mammalian brain exhibits sex differences with respect to structure and function. In our previous report, we found that progranulin (PGRN)-deficient (pgrn(-/-)) mice displayed an alteration in male-type behaviors, including reduced frequency of ejaculation and elevated levels of aggression and anxiety. The aim of the present study was to elucidate the role of PGRN in sex differences in anxiety. In the elevated plus maze, wild-type (pgrn(+/+)) female mice spent more time in the closed arms than the pgrn(+/+) males, suggesting that the level of anxiety was higher in females than males. On the other hand, no sex difference was observed in the pgrn(-/-) mice, and their anxiety levels were almost the same as those of the pgrn(+/+) females. To elucidate the effect of testosterone on male anxiety, male mice were castrated at 5 weeks of age and silastic tubes filled with either testosterone or cholesterol were then implanted into them for one week. These treatments did not affect anxiety in the open field in either genotypes, although the pgrn(-/-) males exhibited higher anxiety than pgrn(+/+) males. Next, we measured the volume of the paraventricular nucleus (PVN) and the locus ceruleus (LC), as these are anxiety/stress-related nuclei that are known to have sex differences in their structures. In the pgrn(+/+) mice, there was a tendency for the volume of the LC to be larger in males than females. In addition, the pgrn(-/-) mice had a larger volume of LC than the pgrn(+/+) mice, although no sexual differences were observed. The number of cells in the LC was also larger in the pgrn(-/-) than in the pgrn(+/+) mice. No significant differences in the volumes of the PVN were observed between genotypes or sexes. These results suggest that PGRN plays a role in organization of the LC, which eventually modulates anxiety in novel environments. PMID- 19550108 TI - Hyperglycemia reduces mitochondrial content and glucose transporter expression in mouse embryos developing in vitro. AB - The objective of this research was to examine the effects of high concentrations of glucose on mouse embryos developing in vitro by studying embryo viability, mitochondrial content and expression of glucose transporters. Addition of 55 mM glucose to the culture medium of two-cell stage embryos significantly reduced the formation of morulae and blastocysts, resulting in fewer cells in the blastocyst stage embryos and increased levels of apoptosis. Quantitative reverse transcriptase (RT) PCR analysis revealed that the expression levels of the pro apoptotic genes Bax and Casp3 at the blastocyst stage were increased significantly by the addition of either 25 or 55 mM glucose to the culture medium. However, addition of 25 or 55 mM glucose to the culture medium did not change the copy numbers of the apoptosis-related miRNAs mmu-mir-15a, mmu-mir-16 and mmu-mir-21. MitoTracker Green fluorescence revealed a decrease in the mitochondrial mass. The expression levels of the mitochondrial DNA-encoded genes Cox1 and Cox2 decreased sharply with the addition of 25 or 55 mM glucose to the culture medium. Both transcripts and protein synthesis of the glucose transporters Glut1 and Glut3 were reduced in blastocysts cultured in the presence of either 25 or 55 mM glucose. These results suggest that hyperglycemia reduces both mitochondrial content and expression levels of glucose transporters in mouse embryos developing in vitro and that this may result in apoptosis in these embryos. PMID- 19550109 TI - Subjecting holstein heifers to stress during the follicular phase following superovulatory treatment may increase the female sex ratio of embryos. AB - The sex ratio of mammals has previously been shown to be affected by maternal stress. In our previous study, the proportion of female embryos collected from superovulated and artificially inseminated Holstein heifers that were frequently placed in stanchions and subjected to transrectal examinations of the ovaries during the follicular phase tended to be higher than the expected 50%. The goal of the present study was to test the validity of this observation using a greater number of heifers. Superovulated heifers were artificially inseminated at 56 and 72 h after PGF(2alpha) treatment using a single batch of frozen semen. Frequent capture (FC), transrectal examination and/or blood sampling were performed at 4-h intervals from 36 to 76 h after PGF(2alpha) treatment (n=13). Nine heifers were used as the Control (non-treatment). Seven-day-embryos were recovered by uterine flushing. Male and female embryos were separated using the loop-mediated isothermal amplification procedure. The proportion of female transferable embryos in the FC group (67.8%, 78/115) was significantly higher than that in the Control group (51.2%, 43/84, P<0.05). The peak concentration of plasma cortisol during the follicular phase following superovulatory treatment was 20.6 ng/ml in the FC group. These results suggest that subjecting heifers to stress during the follicular phase following superovulatory treatment may increase the female sex ratio of embryos. PMID- 19550110 TI - Insufficient amount of Cdc2 and continuous activation of Wee1 B are the cause of meiotic failure in porcine growing oocytes. AB - In mammals, growing oocytes with a diameter less than 80% of that of full-grown oocytes cannot start meiotic maturation, and their maturation promoting factor (MPF) cannot be activated by hormonal stimulation or isolation from follicles. The aim of the present study was to identify the key molecules responsible for meiotic failure of these growing oocytes (referred to as "small oocytes" in the present study). To this end, we altered the expression of the molecules involved in MPF activation in the small oocytes of pigs by injecting them with mRNA or antisense RNA (asRNA) and examined the effects on the meiotic ability of the small oocytes. Immunoblotting analyses revealed three defects in small oocytes compared with full-grown oocytes, an inactive mitogen activated protein kinase (MAPK) cascade, a failure of cyclin B synthesis and an insufficient amount of Cdc2. Injection with mRNAs of Mos, the uppermost molecule of the MAPK cascade, cyclin B1, cyclin B2 or Cdc2 into small porcine oocytes indicated directly and for the first time that the cause of meiotic failure of porcine small oocytes is an insufficient amount of Cdc2 rather than MAPK inactivation or failure of cyclin B synthesis. Next, in order to suppress Myt1 and Wee1B, which phosphorylates at inhibitory phosphorylation sites of Cdc2 and inactive MPF, we injected their asRNAs into the porcine small oocytes and found that the Wee1B asRNA significantly increased meiotic ability, whereas the Myt1 asRNA had no effect. When Cdc2 overexpression and suppression of Wee1B expression were simultaneously induced in the small oocytes of pigs, about 70% of the small oocytes resumed meiosis, and this rate was nearly comparable with that of the full-grown oocytes. These results strongly suggest that an insufficient amount of Cdc2 and continuous activation of Wee1 B are the cause of meiotic failure of small oocytes in pigs. PMID- 19550111 TI - Expression of c-MYC in nuclear speckles during mouse oocyte growth and preimplantation development. AB - Myelocytomatosis oncogene (c-myc) is a major transcriptional regulator that controls various biological processes, and its deregulated expression causes carcinogenesis. To investigate the involvement of c-myc in oogenesis and preimplantation development, the expression of c-MYC during these stages was examined by immunocytochemistry. A strong c-MYC signal was detected in the nucleus of growing and fully grown oocytes as well as in preimplantation embryos before the morula stage. The signal intensity decreased slightly at the morula stage, and no signal was detected in blastocysts. Close observation of the nucleus revealed that c-MYC was localized in small granules that appeared to be nuclear speckles controlling pre-mRNA splicing. Although the number of granules decreased during oocyte growth, their size increased. After fertilization, the granules of c-MYC disappeared from the pronuclei, and c-MYC was evenly distributed in the nucleoplasm at the 1-cell stage, but the granules reappeared at the 2-cell stage. These results suggest that c-myc is involved in oocyte growth and preimplantation development and that its role changes during these stages. PMID- 19550112 TI - [A case of Wernicke-Korsakoff syndrome who succeeded in returning to work despite the sequelae]. PMID- 19550113 TI - Expression of cytochrome P450 arachidonic acid epoxygenase 2J2 in human tumor tissues and cell lines. AB - BACKGROUND AND OBJECTIVE: Cytochrome P450 arachidonic acid epoxygenase 2J2 (CYP2J2) is a new metabolic pathway of arachidonic acid. However, its biological effects, especially pathophysiologic significance in human beings, remain to be further recognized. This study was to determine the expression of CYP2J2 in human tumor tissues and cell lines. METHODS: The expression of CYP2J2 mRNA and protein in 130 specimens of human carcinoma and related adjacent normal tissues, four specimens of inflammatory pseudotumor tissues, eight human tumor cell lines and two normal cell lines (as control) was detected by reverse transcription polymerase chain reaction (RT-PCR), western blot and immunohistochemistry. RESULTS: CYP2J2 was highly expressed in 101 (78%) carcinoma tissues, but was not detected in adjacent normal tissues and inflammatory pseudo-tumor tissues. Its mRNA level was obviously correlated to its protein level (r = 0.613, p < 0.01). Immunohistochemistry analysis showed the same results as RT-PCR and western blot. Furthermore, CYP2J2 was only expressed in cancer cells but not in interstitial and inflammatory cells. CYP2J2 was highly expressed in all carcinoma cell lines, but not in two normal cell lines. CONCLUSION: CYP2J2 is highly and selectively expressed in human tumor tissues and cell lines and may be a novel biomarker of human tumors. PMID- 19550114 TI - GCPReg package for registration of the segmentation gene expression data in Drosophila. AB - In modern functional genomics registration techniques areused to construct reference gene expression patterns and createa spatiotemporal atlas of the expression of all the genes in anetwork. In this paper we present a software package calledGCPReg, which can be used to register the expression patterns ofsegmentation genes in the early Drosophila embryo. The key task,which this package performs, is the extraction of spatially localizedcharacteristic features of expression patterns. To facilitatethis task, we have developed an easy-to-use interactive graphicalinterface. We describe GCPReg usage and demonstrate how thispackage can be applied to register gene expression patterns inwild type and mutants. GCPReg has been designed to operate ona UNIX platform and is freely available via the Internet at http://urchin.spbcas.ru/downloads/GCPReg/GCPReg.htm. PMID- 19550115 TI - Comparative proteomics analysis on differentiation of human promyelocytic leukemia HL-60 cells into granulocyte and monocyte lineages. AB - BACKGROUND AND OBJECTIVE: Human promyelocytic leukemia cell line HL-60 has potential to differentiate into granulocytes and monocytes by different inducers, such as NSC67657 and all-trans retinoic acid (ATRA). However, the mechanism is not clear yet. This study was to induce differentiation of HL-60 cells using ATRA and NSC67657, and compare the protein expression patterns using two-dimensional electrophoresis (2-DE). METHODS: HL-60 cells were cultured with ATRA and NSC67657 respectively. Cell proliferation was analyzed by MTT assay. Cellular surface specific antigen CD11b/CD14 was detected using flow cytometry (FCM) to assess cell differentiation. The alterations of cell morphology were observed with cellular chemical staining under electron microscope. Total protein was separated by modified 2-DE. The differentially expressed proteins were identified using PD Quest software and analyzed by MOLDI-TOF MS. ICAT protein with differential expression was verified by reverse transcription-polymerase chain reaction (RT PCR) and western blot. RESULTS: The granulocytic and monocytic differentiation of HL-60 cells was induced by ATRA and NSC67657, respectively. The positive rates of both CD11b and CD14 in HL-60 cells were over 90% after 5-day treatment (2 micromol/L ATRA or 10 micromol/L NSC67657); cell morphology also represented characteristics of differentiation. Proteomic analysis showed that 25 proteins were differentially expressed with the same pattern in both differentiation groups, ten were differentially expressed only in monocytic differentiation group and 15 only in granulocytic differentiation group. Among them, ICAT expression was upregulated during NSC67657-induced differentiation of HL-60 cells. CONCLUSION: A batch of differentially expressed proteins has be found by 2-DE in HL-60 cells with granulocytic and monocytic differentiation, which would contribute to the following functional research on related proteins. PMID- 19550116 TI - Reversal effect of PI3-K inhibitor LY294002 on P-glycoprotein-mediated multidrug resistance of human leukemia cell line K562/DNR and gastric cancer cell line SGC7901/ADR. AB - BACKGROUND AND OBJECTIVE: Phosphatidylinositol-3-kinase/protein kinase B (PI3 K/Akt) signaling pathway plays an important role in cell survival. This study was to explore the reversal effect of PI3-K inhibitor LY294002 on p-glycoprotein (P gp)-mediated multidrug resistance in human leukemia cell line K562/DNR and gastric cancer cell line SGC7901/ADR. METHODS: The cells were divided into simple drug-treated groups and LY294002 pretreated groups: the former groups received treatment of daunorubicin (DNR), adriamycin (ADR), vincristine (VCR) and etoposide (VP-16), respectively; the latter groups received pretreatment of LY294002 before drug treatment. Trypan blue dye exclusion method and MTT assay were used to detect the drug sensitivity of K562/DNR and SGC7901/ADR cells, and the effect of LY294002 on the drug resistance. The expression of P-gp and phosphorylated Akt (p-Akt) in K562/DNR, SGC7901/ADR and their parental cell lines K562 and SGC7901 was detected by Western blot. Intracellular drug accumulation was measured by flow cytometry (FCM). RESULTS: LY294002 pretreatment significantly decreased the 50% inhibition concentration (IC(50)) of DNR, ADR, VCR and VP-16 for K562/DNR cells, with reverse efficiencies of 72.4%, 64.9%, 60.4% and 52.8%. In SGC7901/ADR cells, the similar result was obtained with a reverse efficiency of 31.0%. LY294002 pretreatment partially inhibited the expression of p-Akt and P-gp, and promoted the intracellular accumulation of DNR and ADR in K562/DNR and SGC7901/ADR cells, respectively. CONCLUSION: LY294002 could partially reverse multidrug resistance in K562/DNR and SGC7901/ADR cells in vitro via inhibiting PI3-K/Akt pathway. PMID- 19550117 TI - Effect of RhoA signaling transduction on expression of Ezrin in breast cancer cell lines. AB - BACKGROUND AND OBJECTIVE: Both RhoA and Ezrin are confirmed to play an important role in the development and metastasis of tumors. However, the mechanism remains unclear. This study rudimentally investigates the regulatory effect of RhoA on the expression of Ezrin. METHODS: After MDA-MB-231 cells were treated with epidermal growth factor (EGF) or following pretreatment of fasudil (the special inhibitor of RhoA), the expression of RhoA, p-RhoA and Ezrin in MDA-MB-231 cells was detected by western blot. RESULTS: Stimulation of EGF triggered RhoA phosphorylation in MDA-MB-231 cells which reached the maximum at 30 min; RhoA expression did not change; Ezrin expression was enhanced and reached the maximum at 24 h. However, pretreatment of fasudil before EGF stimulation decreased RhoA phosphorylation and Ezrin expression in MDA-MB-231 cells by 72.73% and 51.28%, respectively. CONCLUSION: RhoA may regulate the invasion and metastasis of breast cancer cells as an upstream signaling of Ezrin. PMID- 19550118 TI - Inhibitory effect of wild-type p53 gene on excessive replication of centrosomes in leukemia cell line K562. AB - BACKGROUND AND OBJECTIVE: Mutation and deletion of the p53 gene in tumor cells is one of the major reasons for aneuploid development and genomic instability. Abnormal centrosomes exist in chronic myelogenous leukemia patients at different stages; furthermore, the degree of abnormality is associated with the clinical stage and more severe in the blast crisis stage. This study was to establish the leukemia cell line K562 with the exogenous wild-type p53 (wt-p53) gene, and to explore the effect of the p53 gene on centrosomes in K562 cells. METHODS: The recombinant adenoviruses carrying the wt-p53 gene (Ad5wtp53), the mutant p53 gene (Ad5mtp53) and the green fluorescent protein gene (Ad5GFP) were amplified respectively in HEK293 cells, and co-infected with cation polybrene into K562 cells respectively; uninfected K562 cells were used as blank control. The infection efficiency was analyzed by flow cytometry. P53 expression was detected by Western blot. Centrosomes were counted under the laser confocal microscope after indirect immunofluorescence staining. The expression of Gadd45a (growth arrest and DNA damage), BubR1 (Bub 1 related) and Aurora A was detected by western blot. RESULTS: K562 cell line with exogenous wt-p53 gene was established. The infection efficiencies of three groups were over 60%, and P53 sustained expression for 72 h. The percentage of cells with amplified centrosomes (more than 2/cell) in Ad5wtp53 group was decreased to (0.38 +/- 0.02)%, lower than that of blank control group (p < 0.05). Meanwhile, the protein levels of Gadd45a and BubR1 in Ad5wtp53 group were upregulated by 93% and 88% of blank control (p < 0.05), respectively, and the protein level of Aurora A was downregulated by 56% of blank control (p < 0.05). CONCLUSIONS: P53 protein is sustained to express in K562 cells after being infected by Ad5wtp53. wt-p53 can suppress excessive replication of centrosomes that may contribute to the upregulation of Gadd45a and BubR1 protein expression as well as the downregulation of Aurora A protein expression. PMID- 19550119 TI - Effects of arsenic trioxide under different administration ways on T-cell lymphoma xenografts in nude mice: in vivo and in vitro experiments. AB - BACKGROUND AND OBJECTIVE: In vitro, arsenic trioxide (As(2)O(3)) can inhibit proliferation of many lymphoma cell lines. In clinic, it also can be used to treat many subtypes of lymphoma. But the dosage and administration ways are undetermined yet. In this research, we studied the antitumor effect of As(2)O(3) with different administration ways on T-cell lymphoma and observed the toxicity. METHODS: Murine T-cell lymphoma cell line EL4 was treated with As(2)O(3) of eight concentrations. Cell proliferation was detected by MTT assay. Cell apoptosis was evaluated by flow cytometry with Annexin-V-FITC/PI staining and observed under electroscope and fluorescent microscope. EL4 cells were inoculated into nude mice to establish lymphoma models. The effect of As(2)O(3) on lymphoma in nude mice was observed. RESULTS: As(2)O(3) inhibited the proliferation of EL4 cells with a 50% inhibition concentration (IC(50)) of 1.28 micromol/L at 72 h (p < 0.05). When treated with the same total dose of As(2)O(3) by 4 mg(kg d)(-1) for seven days or 2 mg(kg d)(-1) for 14 days, the inhibition rates of tumor growth in mice were equivalent (58.8% vs. 55.6%, p = 0.351). Apoptotic cells were increased and apoptotic bodies were observed in xenograft tumor tissues. Acute liver damage is the major toxicity. CONCLUSION: Shortening the administration course and increasing the daily dosage of As(2)O(3) can be considered as a reasonable model for treating advanced/refractory lymphomas. PMID- 19550120 TI - Inhibitory effect of shRNA targeting GPR48 on invasion and metastasis of human cervical carcinoma cell line HeLa. AB - BACKGROUND AND OBJECTIVE: GPR48 regulates invasion and metastasis of cancer cells by linking Gs protein to stimulate intracellular signaling pathway and affect actin filament aggregation and the activity of matrix metalloproteinase. This study was to explore the effects of GPR48 short hairpin RNA (shRNA) on GPR48 expression, and invasion and metastasis of cervical carcinoma cell line HeLa. METHODS: Plasmids containing two different sequences of human GPR48 mRNA coding region were constructed and transfected into HeLa cells. HeLa cell clones with stable small interfering RNA (siRNA) expression were screened by neomycin resistance. The expression of GPR48 was detected by reverse transcription polymerase chain reaction (RT-PCR) and western blot. In vitro invasive activity was measured by Boyden chamber experiment. GPR48 shRNA-transfected or untransfected HeLa cells were inoculated subcutaneously into nude mice to observe tumor growth and lung metastasis. RESULTS: Compared with that in negative shRNA transfected cells, GPR48 mRNA expression in GPR48 shRNA-transfected cells was downregulated by more than 80% and the number of HeLa cells infiltrated Boyden chamber membrane was decreased (94.4 +/- 15.7 vs. 28.3 +/- 1.5 and 17.6 +/- 1.5, p < 0.01). The depletion of endogenous GPR48 suppressed in vivo metastasis: the number of lung metastases was significantly larger in negative shRNA group than in GPR48 shRNA groups (7.3 +/- 1.8 vs. 1.3 +/- 0.3 and 1.5 +/- 0.4, p < 0.01). CONCLUSION: GPR48 shRNA can inhibit in vitro invasion and in vivo metastasis of HeLa cells efficiently. PMID- 19550121 TI - Correlation of Notch1 expression and activation to cisplatin-sensitivity of head and neck squamous cell carcinoma. AB - BACKGROUND AND OBJECTIVE: Notch1 belongs to the Notch family of transmembrane receptors and plays an important role in tumor cell proliferation and apoptosis. Notch1 affects chemosensitivity of tumors. However, its correlation to cisplatin (DDP)-sensitivity of head and neck squamous cell carcinoma is unclear. This study was to identify the expression of Notch1 in head and neck squamous cell carcinoma, and investigate its influence on the DDP-sensitivity. METHODS: Twenty five fresh specimens of head and neck squamous cell carcinoma were subjected to primary cell culture. DDP-sensitivity of tumor cells was detected using collagen gel droplet embedded culture-drug sensitivity test (CD-DST). The expression of Notch1 in head and neck squamous cell carcinoma, normal squamous epithelium, and tongue squamous cell carcinoma Tb3.1 cells was detected by immunohistochemistry or immuocytochemistry. Tb3.1 cells were divided into four groups, and received treatment of DMSO, DAPT, DMSO plus DDP, DAPT plus DDP, respectively. The absorbance of the four groups was detected by CD-DST to evaluate DDP-sensitivity. RESULTS: The positive rate of Notch1 was significantly higher in head and neck squamous cell carcinoma than in normal squamous epithelium (100% vs. 35%, p < 0.001), and it was negatively correlated to DDP-sensitivity (r = -0.705, p < 0.01). There was no difference in absorbance between DMSO group and DAPT group (155.4 +/- 2.3 vs. 154.7 +/- 1.2, p > 0.05), while the absorbance was significantly higher in DMSO plus DDP group than in DAPT plus DDP (33.9 +/- 1.3 vs. 26.6 +/- 1.1, p < 0.05). CONCLUSIONS: Notch1 expression is negatively correlated to DDP-sensitivity of head and neck squamous cell carcinoma, and could be used to predict DDP-sensitivity. DAPT can enhance DDP-sensitivity of Tb3.1 cells via blocking Notch1 signaling. PMID- 19550122 TI - Caspase-dependent molecular mechanisms of anti-human DR5 monoclonal antibody mDRA 6 inducing apoptosis of human leukemia Jurkat cells. AB - BACKGROUND AND OBJECTIVE: Both tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and some monoclonal agonistic antibodies against TRAIL receptors have antitumor activity. We have previously prepared a novel monoclonal agonistic antibody against human death receptor 5 (DR5) and designated it as mDRA-6. This study was to explore the Caspase-dependent molecular mechanisms of mDRA-6 inducing apoptosis of human leukemia Jurkat cells. METHODS: After exposure to different doses of mDRA-6, DNA fragmentation of Jurkat cells was detected by agarose gel electrophoresis, cell proliferation was detected by MTT assay, and cell apoptosis was detected by flow cytometry after Annexin V-FITC/PI double staining. Jurkat cells were further treated with the inhibitors for Caspase-10, 9, -8 and -3. The active cleavage products of Caspase-10, -9, -8, -3 and poly ADP ribose polymerase (PARP), BH3 interacting domain death agonist (Bid), truncated Bid (tBid) and cytochrome c (Cyto c), were analyzed by western blot. RESULTS: After mDRA-6 treatment, DNA fragmentation was detected in Jurkat cells. mDRA-6 inhibited cell proliferation in a dose-dependent manner. When treated with 2.0 microg/mL mDRA-6, the apoptosis rates of Jurkat cells were 16.2% at 0.25 h, 28.3% at 0.5 h, 69.2% at 1 h and 78.2% at 2 h. Interestingly, the mDRA-6-induced apoptosis was repressed by 77.9% by Caspase-8 inhibitor ZIF, 54.2% by Caspase-3 inhibitor ZDF, and 8.7% by Caspase-9 inhibitor ZLF, but was not repressed by Caspase-10 inhibitor ZAF. After mDRA-6 exposure, the proenzymes of Caspase-8, -9 and -3 were reduced and their active cleavage products were increased along with the increase of exposure time, the cleavage products of PARP were also increased, Bid was degraded to tBid, and an abundance of Cyto c was released from mitochondria, but the proenzyme of Caspase-10 showed no change and no cleavage products of Caspase-10 were detectable. CONCLUSION: mDRA-6 can induce apoptosis of Jurkat cells via the Caspase-dependent and mitochondrial pathways. PMID- 19550123 TI - Biological characteristics of dendritic cells derived from peripheral blood of patients with epithelial ovarian cancer. AB - BACKGROUND AND OBJECTIVE: Dendritic cells (DCs) are thought to be the most potent antigen-presenting cells (APC) and play a vital role in stimulating human immune response against cancer. At present, most data concerning the immuno-biological function of DCs are obtained from healthy donors. The information about the biological characteristics of DCs from patients is limited. In this study, the biological characteristics of monocyte-derived dendritic cells (MoDCs) from patients with ovarian cancer were investigated. METHODS: Monocytes were isolated from peripheral blood mononuclear cells (PBMC) of eight epithelial ovarian cancer patients and 13 healthy women volunteers, cultured with interleukin 4 (IL-4) and granulocyte-macrophage colony stimulating factor (GM-CSF), and stimulated with tumor necrosis factor-alpha (TNF-alpha). At seven days after induction, the morphologic characteristics of MoDCs were observed. The features of phenotype were analyzed using flow cytometry. The ability of MoDCs to stimulate proliferation of lymphocytes was tested by allogeneic mixed leukocytes reaction (MLR). RESULTS: Mature MoDCs with typical morphology were obtained after seven days of culture. MoDCs from both patients and healthy women expressed high levels of HLA-ABC (MHC-I), HLA-DR (MHC-II) and large amounts of CD86 and CD80. There was no significant differences between MoDCs from ovarian cancer women and healthy women in the mean fluorescence intensity (MFI) of HLA-ABC, HLA-DR, CD86 and CD80 (p > 0.05). The MLR was significantly weaker in ovarian cancer patients than in healthy women (p < 0.05). CONCLUSION: MoDCs from ovarian cancer patients may present lower capacity of stimulating proliferation of lymphocytes, indicating that the patients' MoDCs may have immunological function defect at certain extent. PMID- 19550124 TI - Effect of diallyl disulfide on cell cycle arrest of human colon cancer SW480 cells. AB - BACKGROUND AND OBJECTIVE: Our previous study revealed that diallyl disulfide (DADS) significantly inhibited cell proliferation and induced cell cycle arrest at G(2)/M phase of human colon cancer SW480 cells. However, the molecular mechanism of cell cycle arrest remains unclear. This study was to investigate the role and the molecular mechanism of DADS in the induction of cell cycle arrest of human colon cancer cell line SW480. METHODS: Proliferation of SW480 cells after DADS treatment was measured by MTT assay and cell counting. Phase distribution of cell cycle was analyzed by flow cytometry. Expressions of PCNA, p53, p21(WAF1) and cyclin B1 were detected by immunohistochemistry and western blot. RESULTS: DADS (30-70 microg/mL) significantly inhibited proliferation and retarded the population doubling time of colonies in SW480 cells. Compared with the control group, SW480 cells were markedly accumulated at G(2)/M phase after the treatment with DADS (p < 0.05). Moreover, DADS remarkably decreased the protein contents of PCNA, p53 and cyclin B1, but increased the expression of p21(WAF1) in a time- and dose-dependent manner. CONCLUSION: DADS induces G(2)/M arrest in human colon cancer SW480 cells, probably through the downregulation of PCNA, p53 and cyclin B1 and upregulation of p21(WAF1). PMID- 19550125 TI - Expression and clinical significance of YKL-40 protein in epithelial ovarian cancer tissues. AB - BACKGROUND AND OBJECTIVE: Overexpression of YKL-40 has been detected in the sera from patients with various kinds of malignant tumors, including epithelial ovarian cancer. Moreover, YKL-40 expression is closely related to clinical phenotypes of some malignant tumors. This study was to investigate the expression and clinical significance of YKL-40 protein in epithelial ovarian cancer tissues. METHODS: Protein expression of YKL-40 was detected by immunohistochemistry (IHC) using tissue microarray (TMA) consisting of 86 specimens of epithelial ovarian cancer and 20 specimens of normal ovarian tissues. The correlations of YKL-40 expression to clinical features and prognosis, as well as to the expression of clusterin protein in epithelial ovarian cancer were evaluated. RESULTS: The expression of YKL-40 in all normal ovarian tissues was negative or at low levels. In 74 evaluable specimens of epithelial ovarian cancer, overexpression of YKL-40 was detected in 42 cases (56.8%). YKL-40 expression was closely associated with the clinical stage of epithelial ovarian cancer (p < 0.0001). The overall survival time in patients with overexpression of YKL-40 was significantly shorter than that in patients with normal expression of YKL-40 (p = 0.0389). Moreover, expression of YKL-40 protein was positively correlated with that of clusterin protein in epithelial ovarian cancer (p < 0.0001). CONCLUSION: YKL-40 may be used as a new molecular marker to predict the prognosis of epithelial ovarian cancer. PMID- 19550126 TI - Short-term efficacy of rituximab-CHOP and CHOP regimens on two subtypes of diffuse large B-cell lymphoma. AB - BACKGROUND AND OBJECTIVE: Diffuse large B-cell lymphoma (DLBCL) can be divided into two subgroups as germinal center B-cell like (GCB) and non-GCB DLBCL, according to the origin of tumor cells. The prognosis of GCB DLBCL is better than that of non-GCB DLBCL after receiving the CHOP regimen on initial therapy. This study was to compare the short-term efficacy of rituximab(R)-CHOP and CHOP regimens on GCB and non-GCB DLBCL, thus to explore the optimal first-line regimen for the initial treatment of DLBCL patients. METHODS: In total, eighty-three patients with de novo DLBCL initially treated at Sun Yat-sen University Cancer Center from November 2006 to February 2008 were enrolled. Patients were divided into GCB and non-GCB groups. The short-term efficacy of the CHOP or R-CHOP regimen as the first-line therapy on the two groups was evaluated according to the revised response criteria for lymphoma. Bcl-2 expression and its correlation to the short-term efficacy of the two groups were assessed. RESULTS: There were 35 cases (42.2%) in the GCB group and 48 cases (57.8%) in the non-GCB group. The total remission rate was 74.3% in the GCB group and 60.4% in the non-GCB group (p = 0.006). There was no significant difference in Bcl-2 expression in the two groups. The remission rate was higher in Bcl-2 positive patients receiving R-CHOP regimen than those receiving CHOP regimen (75.6% vs. 47.8%, p = 0.023). There was no significant difference in the remission rate of Bcl-2 negative patients, regardless of chemotherapy regimen. CONCLUSION: The prognosis is better in the GCB group than in the non-GCB group. Addition of rituximab to CHOP could improve the short-term efficacy of Bcl-2 positive patients. PMID- 19550127 TI - Insulin resistance in patients with advanced prostate cancer undergoing intermittent or continuous androgen blockade. AB - BACKGROUND AND OBJECTIVE: Androgen blockade is the principle strategy in the treatment of advanced prostate cancer. Impaired glucose tolerance often occurs in those patients after androgen blockade. This study was to investigate the correlation of insulin resistance to intermittent androgen blockade (IAB) or continuous androgen blockade, which is also named surgical castration, in patients with advanced prostate cancer. METHODS: A total of 139 patients with advanced prostate cancer were classified into four groups according to the body mass index (BMI) and the treatment method. Group A consisted of 30 patients receiving surgical castration with BMI >or= 24 kg/m(2), group B consisted of 32 patients treated with IAB with BMI >or= kg/m(2), group C consisted of 37 patients undergoing surgical castration with BMI < 24 kg/m(2), group D consisted of 40 patients treated by IAB with BMI < 24 kg/m(2). Fasting plasma glucose (FPG) and fasting serum level of insulin (FINS) were assessed before treatment, six months and 12 months after treatment, respectively. Insulin resistance index (IRI) was also calculated. RESULTS: Six months after treatment, FINS and IRI were all increased in the four groups compared with those before treatment; FINS and IRI were significantly higher in groups B and D than in A and C (FINS: t(A:B)=7.7516, p < 0.01, t(C:D)=4.8078, p < 0.01; IRI: t(A:B) =7.3671, p < 0.01, t(C:D)=5.1005, p < 0.01). Twelve months after treatment, which was the intermittent period of the IAB method, FINS returned to the pretreatment level in group D (q=2.5255, p > 0.05), and dramatically decreased in group B compared to the value six months after treatment (q = 9.0942, p < 0.01); in contrast, FINS and IRI remained unchanged in groups A and C. CONCLUSIONS: Androgen blockade promotes insulin resistance in patients with advanced prostate cancer. Insulin resistance gradually disappears during the intermittent period of IAB. PMID- 19550128 TI - Expression of CXCL12/CXCR4 and its correlation to prognosis in esophageal squamous cell carcinoma. AB - BACKGROUND AND OBJECTIVE: CXCL12/CXCR4 is expressed in many kinds of tumors, which is associated with tumor proliferation and invasion. This study was to investigate the expression of CXCL12/CXCR4, and explore its correlation to prognosis and clinicopathologic factors of esophageal squamous cell carcinoma (ESCC). METHODS: The expression of CXCL12/CXCR4 protein in 186 specimens of ESCC was assessed by immunohistochemistry. RESULTS: The positive rates of CXCL4 and CXCR12 protein in ESCC tissues were 67.2% and 63.4%, respectively. CXCL4 and CXCR12 were not expressed in 20 specimens of normal esophageal epithelium. PTNM stage and positive expression of CXCR4 were independent prognostic factors of ESCC (p < 0.05). The five-year survival rates of CXCL12-positive and CXCL12 negative groups were not significantly different (21.0% vs. 18.8%, p > 0.05), while the five-year survival rate was significantly higher in CXCR4-negative group than in CXCR4-positive group (28.5% vs. 2.2%, p < 0.05). The expression of CXCR4 was higher in the group with lymph node metastasis and pathological T3 stage than in the group without lymph node metastasis and with pathological T1-T2 stages (p < 0.05). The expression of CXCR4 was not correlated with the expression of CXCL12 in ESCC. CONCLUSIONS: CXCL12/CXCR4 is intensively expressed in esophageal squamous cell carcinoma. The level of CXCR4 is positively correlated to progression and prognosis of ESCC. PMID- 19550129 TI - Application of multislice spiral CT angiography on transcatheter arterial chemoembolization for hepatocellular carcinoma. AB - BACKGROUND AND OBJECTIVE: Multislice spiral CT angiography (MSCTA) is very important in the diagnosis and treatment of liver diseases. Currently, most studies on three-dimensional MSCTA of the liver vascular system focus on the liver tumors, preoperative assessment of liver transplantation and the systematic anatomy of the liver vascular system. This study was to investigate the clinical application of MSCTA on transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) by comparing images of MSCTA and digital subtraction angiography (DSA). METHODS: MSCT dual-phase enhanced scanning was performed in 50 patients with advanced HCC. Both hepatic artery angiography and portal vein angiography were conducted using maximal intensity projection (MIP) and volume rendering technique (VRT). DSA of the celiac artery, superior mesenteric artery, renal artery and diaphragm artery, as well as TACE were performed in all patients. MSCTA and DSA images of the 50 patients were compared. RESULTS: MSCTA and DSA showed equal detectability in revealing classification of the hepatic artery anatomy and tumor blood vessels, with a coincidence of 100% (p = 1.00). However, MSCTA was superior to DSA in displaying arterioportal shunt and portal vein tumor thrombus. CONCLUSIONS: As a noninvasive and easy to conduct technique, MSCTA can accurately provide information of the hepatic artery, portal vein and tumor supply vessels. Therefore MSCTA has a favorable value to guide TACE for HCC. PMID- 19550130 TI - Dynamic enhancement patterns of solitary pulmonary nodules at multi-detector row CT and correlation with vascular endothelial growth factor and microvessel density. AB - BACKGROUND AND OBJECTIVE: Differential diagnosis of malignant solitary pulmonary nodules (SPNs) from benign ones is difficult based on imaging manifestations. This study was to assess dynamic enhancement patterns of SPNs detected with multi detector row computed tomography (MDCT), correlate SPN manifestations of MDCT to the expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD), thus to explore the potential value of MDCT imaging in the diagnosis of SPNs. METHODS: Fifty pathologically and one clinically confirmed patients with SPNs (diameter 0.05). CONCLUSIONS: Expression of connexin 32 is closely correlated to the differentiation of NSCLC and affects the prognosis of NSCLC patients. Increasing the expression of connexin 32 may improve the prognosis of NSCLC. PMID- 19550133 TI - Application of logistic regression in combination with multiple diagnostic tests for auxiliary diagnosis of nasopharyngeal carcinoma. AB - BACKGROUND AND OBJECTIVE: Although there are many markers for the clinical diagnosis of nasopharyngeal carcinoma (NPC), the efficacy of most of the markers for the early diagnosis is poor. This study was to evaluate the diagnostic value of VCA/IgA, EA/IgA, EBV DNA, EBNA1/IgA, EBNA1/IgG and ZTA/IgG for NPC, as well as to screen out an optimized combination using the logistic regression to increase diagnostic accuracy of NPC. METHODS: Eight-one newly pathologically diagnosed NPC patients prior to treatment and 89 health cases from routine physical checkups were entered into the study. Epstein-Barr virus (EBV) DNA was detected by quantitative real-time PCR; VCA/IgA and EA/IgA were assessed by immunofluorescence assays (IFA). The receiver operating characteristic (ROC) curve and the area under the curve were used to evaluate the diagnostic value of a single test or combined tests for NPC, thus to decide the cut-off value. The logistic regression model was used to combine the results from multiple tests to increase diagnostic accuracy. RESULTS: Comparing to the routine parallel sequential test, the logistic regression in combination with multiple diagnostic tests achieved higher diagnostic specificity and sensitivity for NPC. Two optimal combinations were EBV DNA + EBNA1/IgA and VCA/IgA + EBNA1/IgA, whose sensitivity and specificity reached 0.96 and 0.82, 1.00 and 0.84, respectively. When the logistic model was used and the cut-off value was determined by ROC, the sensitivity and specificity of the two combination groups became 1.00 and 0.87, 0.98 and 0.88, respectively. CONCLUSION: Adopting the logistic regression in combination with multiple diagnostic tests and using the probability prediction to decide the cut-off value may help increase the diagnostic sensitivity and specificity for NPC. PMID- 19550135 TI - Interaction of indoleamine-2,3-dioxyagnase and CD4+CD25+ regulatory T cells in tumor immune escape. AB - There are numerous factors involved in the tumor evasion from immune surveillance. Recently, considerable attention has been given to indoleamine 2,3 dioxygenase (IDO) and CD4(+)CD25(+) regulatory T cells (Tregs). The interaction between the two may play a crucial role in tumor immune escape. This article reviews the correlation between IDO and Tregs in immune escape of tumors. PMID- 19550134 TI - Establishing SCID mouse models of B-cell non-Hodgkin's lymphoma. AB - BACKGROUND AND OBJECTIVE: Recently, the incidence of non-Hodgkin's lymphoma (NHL) is increasing, in which most are aggressive. It is limited for promoting the efficacy of conventional chemotherapy on NHL. In this study, mouse models of B cell NHL were established for determining the efficacy and mechanisms of novel therapies. METHODS: Diffuse large B-cell lymphoma SU-DHL-4 cells and Burkitt's lymphoma Daudi cells were injected into SCID (severe combined immunodeficiency) mice through the tail veins to observe the presentations and requirements for establishing mouse models. The Daudi-cell lymphoma mice were divided into control group and rituximab group, and the latter received treatment of rituximab. The tumor onset and survival time of mice were investigated. RESULTS: The median onset time of SU-DHL-4-cell lymphoma in SCID mice was 39.5 days, which presented cachexia, weight loss, erect hair, tardiness and enlarged tumors in the abdomen, rump or pelvic limb, but without tumor cell infiltration in the liver, spleen or bone marrow. The median onset time of Daudi-cell lymphoma in SCID mice was 30.5 days, which were characterized by paralyzed lower limbs and died about 9.5 days after paralysation. Most organs such as the liver, kidney, spleen and bone marrow were infiltrated by a number of Daudi cells. After treatment of rituximab, Daudi cells presented typical characteristics of apoptosis. The median paralysis time and survival time of mice with Daudi-cell lymphoma were significantly longer in rituximab group than in control group (52.5 days vs. 30.5 days, 76.5 days vs. 40 days, p < 0.05). CONCLUSION: SCID mouse models of B-cell lymphoma can be successfully established with either SU-DHL-4 cells or Daudi cells. PMID- 19550136 TI - Site-specific metastasis formation: chemokines as regulators of tumor cell adhesion, motility and invasion. AB - The metastatic spread of tumors is a well-coordinated process in which different types of cancers tend to form metastases in defined organs. The formation of site specific metastases requires full compatibility between the intrinsic properties of the tumor cells and the tumor microenvironment. It was recently found that chemokines which are expressed in specific loci promote the adhesion, migration and invasion of tumor cells that express the corresponding receptor(s). Of the different members of the family, the CXCL12 chemokine and its cognate CXCR4 receptor are the prototypes of this process, although other members of the family (e.g. CCR7 and CCR10) also play a role in determination of the metastatic spread. This commentary addresses the fundamental roles of chemokines and their receptors in site-specific metastasis, with emphasis on CXCL12-CXCR4. The article also describes some of the efforts that were performed thus far in order to identify the intracellular components involved in this process. The focus is put on the roles played by proteins that regulate adhesion and migration of tumor cells in response to CXCL12, including mainly focal adhesion kinase (FAK), Pyk2/RAFTK and members of the Rho family of GTPases (RhoA, Rac, Cdc42). This is followed by discussion of open questions that need to be addressed in future research, and of the potential therapeutic implications of the findings that are available to date in this field. PMID- 19550137 TI - Wnt asymmetry and the terminal division of neuronal progenitors. PMID- 19550138 TI - Protein kinase D1: a novel regulator of actin-driven directed cell migration. PMID- 19550139 TI - DNA-PK: relaying the insulin signal to USF in lipogenesis. PMID- 19550140 TI - Frequent epigenetic inactivation of the SLIT2 gene in chronic and acute lymphocytic leukemia. AB - Recently a mouse model of T/natural killer acute lymphoblastic leukemia was used to assess global promoter methylation across the mouse genome using the restriction landmark genomic scanning technique. One of the methylated mouse genes identified in this way was Slit2. There are three mammalian SLIT genes (SLIT1, SLIT2, SLIT3), that belong to a highly conserved family of axon guidance molecules. We have previously demonstrated that SLIT2 is frequently inactivated in lung, breast, colorectal and glioma tumors by hypermethylation of a CpG island in its promoter region, whilst inactivating somatic mutations are rare. Furthermore, we demonstrated that SLIT2 acts as a tumor suppressor gene in breast and colorectal cancer cells. In this report we determined the methylation status of the SLIT2 gene in leukemias (CLL and ALL). SLIT2 was methylated in all ten leukemia cell lines analyzed (eight completely and two partially methylated). SLIT2 expression was restored after treating ALL lines with 5-aza-2dC. In primary ALL and CLL samples, SLIT2 was also frequently methylated, 58% (30/52) B-ALL; 83% (10/12) T-ALL and in 80% (24/30) CLL. Whilst DNA from peripheral blood and bone marrow from healthy control samples showed no SLIT2 methylation. Methylation results in leukemia cell lines and ALL and CLL primary samples were confirmed by direct sequencing of bisulfite modified DNA. Our results demonstrate that methylation of the SLIT2 5' CpG island is conserved between mice and humans, and therefore is likely to be of functional importance. PMID- 19550141 TI - Chronic inflammation, the tumor microenvironment and carcinogenesis. AB - Chronic inflammation often precedes or accompanies a substantial number of cancers. Indeed, anti-inflammatory therapies have shown efficacy in cancer prevention and treatment. The exact mechanisms that turn a wound healing process into a cancer precursor are topics of intense research. A pathogenic link has been identified between inflammatory mediators, inflammation related gene polymorphisms and carcinogenesis. Animal models of cancer have been instrumental in demonstrating the diversity of mechanisms through which every tumor compartment and tumor stage may be affected by the underlying inflammatory process. In this review, we focus on the interaction between chronic inflammation, tumor stem cells and the tumor microenvironment. We summarize the proposed mechanisms that lead to the recruitment of bone marrow derived cells and explore the genetic and epigenetic alterations that may occur in inflammation associated cancers. PMID- 19550142 TI - NADPH oxidases contribute to autophagy regulation. AB - Reactive oxygen species (ROS) are emerging as regulators of autophagy in various cellular contexts. There are many cellular sources of ROS in eukaryotic cells. In phagocytes, the critical immune cells for host defense, the Nox2 NADPH oxidase generates ROS during phagocytosis and plays a central role in microbial killing. Toll-like receptors (TLRs) are important membrane microbial sensing receptors, which can activate Nox2,(1) and were recently demonstrated to signal autophagy targeting of phagosomes to promote their maturation.(2) Our recent study reveals that Nox2 activity and its generated ROS are key signals that induce TLR activated autophagy of phagosomes. Our results provide the first evidence that ROS from the Nox2 NADPH oxidase can contribute to regulating autophagy in host defense against bacteria. The association of TLR, Nox2 and autophagy with inflammatory bowel disease (IBD) suggests a significant role of this antibacterial pathway in these diseases. PMID- 19550143 TI - 1-O-Hexadecyl-2-O-methyl-3-O-(2'-acetamido-2'-deoxy-beta-D-glucopyranosyl)-sn glycerol (Gln) induces cell death with more autophagosomes which is autophagy independent. AB - Autophagic cell death has recently received a great deal of attention. However, a dependence of this type of cell death on the actual process of autophagy has only rarely been proven. Indeed, it is important to differentiate between cell death with an accumulation of autophagosomes and cell death actually caused by excessive or inhibited autophagy. The aim of this study was to elucidate the mechanism of action involved in the cytotoxicity of 1-O-hexadecyl-2-O-methyl-3-O (2'-acetamido-2'-deoxy-beta-D-glucopyranosyl)-sn-glycerol (Gln) and specifically the involvement of autophagy in the effects observed. Our results show that Gln induces cell death associated with large increases in autophagolysosome number and size. However the cell death is independent of autophagy and caspase activation. Instead, Gln leads to lysosomal membrane permeabilization with a resulting leakage of hydrolases into the cytosol, which are then directly involved in cell death. The increased number of autophagolysosomes, however, is just a side effect of the neutralization of the lysosomal pH by Gln. PMID- 19550144 TI - How Yersinia pestis becomes a foreign obstruction in the digestive system of the macrophage. PMID- 19550145 TI - A microarray-based DNA methylation study of glioblastoma multiforme. AB - Glioblastoma multiforme (GBM) is the most frequent and devastating primary brain tumor in adults. The presence of epigenetic lesions, like hypermethylation of known tumor suppressor genes such as MGMT, has been widely described in GBM, but to our knowledge, a genome-wide profile of DNA methylation changes in these lethal tumors is not yet available. In the present analysis, we have quantified the DNA methylation level of 1,505 CpG dinucleotides (807 genes) in 87 consecutive GBMs using universal BeadArrays. Supervised cluster analyses identified 25 and seven genes that were respectively hypermethylated and hypomethylated in more than 20% of the cases studied. The most frequently hypermethylated genes were HOXA11, CD81, PRKCDBP, TES, MEST, TNFRSF10A and FZD9, being involved in more than half of the cases. Studying the biological features of hypermethylated genes, we found that the group of genes hypermethylated in GBM was highly enriched (41%, p < 0.001) for targets of the PRC2 (Polycomb repressive complex 2) in embryonic stem cells. This suggests that GBM might be derived from precursor cells with stem cell-like features. DNA methylation profiles were associated with overall survival in GBM, and we confirmed the favorable prognostic impact of MGMT methylation in patients treated with alkylating agents. Furthermore, we identified that promoter hypermethylation of the transcription factor gene GATA6 (occurring in 30% of GBM) was significantly associated with unfavorable patient survival. PMID- 19550146 TI - Binding Rubicon to cross the Rubicon. AB - Beclin 1 is an antitumor protein, required for mammalian autophagy, but its precise molecular function is poorly understood. Mass spectrometry analysis reveals that two novel proteins, Atg14L and Rubicon, associate with Beclin 1, together with a known Beclin 1-binding protein, UVRAG. The interactions of Atg14L and UVRAG with the Beclin 1-Vps34 (class III PI3-kinase)-Vps15 core complex are mutually exclusive; Rubicon associates with a subpopulation of UVRAG-containing complexes. The Atg14L complex, which positively regulates autophagy at an early step, localizes to the phagophore/isolation membrane, autophagosome and endoplasmic reticulum. In contrast, the Rubicon-UVRAG complex localizes to the late endosome/lysosome and negatively regulates both autophagy at a later step and the endocytic pathway. Thus, the Beclin 1-Vps34-Vps15 complex functions in autophagy and the endocytic pathway, but its function in a given context depends on the identity of its interacting subunits. PMID- 19550147 TI - Autophagic degradation of nuclear components in mammalian cells. AB - Autophagy is an evolutionarily conserved intracellular mechanism for the degradation of organelles and proteins. Here we demonstrate the presence of perinuclear autophagosomes/autolysosomes containing nuclear components in nuclear envelopathies caused by mutations in the genes encoding A-type lamins (LMNA) and emerin (EMD). These autophagosomes/autolysosomes were sometimes bigger than a nucleus. The autophagic nature is further supported by upregulation of LC3-II in Lmna(H222P/H222P) fibroblasts. In addition, inhibition of autophagy led to the accumulation of nuclear abnormalities and reduced cell viability, strongly suggesting a beneficial role of autophagy, at least in these cells. Similar giant autophagosomes/autolysosomes were seen even in wild-type cells, albeit rarely, implying that this "nucleophagy" is not confined to the diseased condition, but may be seen even in physiologic conditions to clean up nuclear wastes produced by nuclear damage. PMID- 19550148 TI - A new therapeutic strategy against cancer: cAMP elevating drugs and leptin. PMID- 19550149 TI - Hyperglycemia, intracellular hyaluronan synthesis, cyclin D3 and autophagy. AB - Hyperglycemia is one of the factors that induces autophagy. Our recent studies demonstrate that dividing cells in hyperglycemic medium initiate an intracellular stress response that involves synthesis of hyaluronan and its extrusion extracellularly into structures that are recognized by inflammatory cells. During the later phase, a complex with cyclin D3, CDK4 and C/EBPalpha was observed in the hyperglycemic cultures, and cyclin D3 and C/EBPalpha colocalized in coalesced perinuclear honeycomb-like structures with embedded hyaluronan. Further, microtubule-associated protein 1 light chain 3 (LC3), a marker for autophagy, colocalizes with these structures. These results suggest that cyclin D3 is a central coordinator that controls the organization of a complex set of proteins that regulate autophagy and subsequent formation of the monocyte-adhesive hyaluronan matrix. However, the early intracellular accumulation of hyaluronan could have a critical role in initiating or regulating these downstream events. PMID- 19550150 TI - Recruitment of 9-1-1 to sites of DNA damage through cell cycle-dependent processing of DNA lesions. PMID- 19550151 TI - Stat1 phosphorylation is a molecular switch of Ras signaling and oncogenesis. PMID- 19550152 TI - Comparative genomics, evolution and origins of the nuclear envelope and nuclear pore complex. PMID- 19550153 TI - Genomic instability induced by heat-inactivated bacteria: implication to tumorigenesis. PMID- 19550154 TI - The involvement of CaM-KII in insulin induced cell proliferation. PMID- 19550155 TI - New life for an "old" drink. PMID- 19550156 TI - Apollo: a healer of the genome? PMID- 19550157 TI - "Misinterpretation" of a histone mark is linked to aberrant stem cells and cancer development. PMID- 19550158 TI - A missing link: the influence of societal beliefs on integration among the world's emerging population of persons with disabilities (PWDs). PMID- 19550159 TI - Disability: a view from selected cultural perspectives. AB - The concept of disability has been examined from various cultural perspectives across the globe. The author explores how services and specific support networks have been provided to people with disabilities. Small-scale and large-scale societies have had to deal with unique challenges in monitoring services for the disabled, regardless of their geographical location; financial resources, accountability and adequate staffing are primary problems in delivery of services to the disabled. Prospects and challenges facing people with disabilities require policy reforms and advancements in special education in all countries. PMID- 19550160 TI - Pregnancy intention and antenatal care use in two rural north Indian States. AB - OBJECTIVES: Many studies have shown strong effects of pregnancy intention on antenatal care (ANC) behaviour in developed countries, but studies from developing settings have shown mixed results. Few investigators have utilized a prospective measure of pregnancy intention. This paper will analyze the association of pregnancy intention and the utilization of antenatal services in two states in northern India, using a prospective measure of whether a future pregnancy would be wanted or unwanted. METHODS: A prospective cohort study was conducted between 1998 and 2003 in Jharkhand and Bihar, India, of 2028 women with one or two pregnancies resulting in the live births of singleton infants during the study period. RESULTS: Antenatal care utilization was not found to be significantly associated with prospective pregnancy intention (OR=1.18 [95% CI 0.91, 1.52]). Among women who received ANC (N = 701), initiation of care was not delayed in unwanted pregnancies. Significant differences existed between the numbers of women who reported their pregnancy unwanted retrospectively compared with prospectively. These differences were not associated with the utilization of antenatal care services or timing of care initiation. The exception to these findings were women who consistently reported their pregnancies unwanted both before and after conception, who were twice as likely to delay ANC initiation as women with consistently wanted pregnancies. CONCLUSIONS: Demographic characteristics of reproductive-age women, such as age and parity, seem to predict more closely the use of ANC services than pregnancy intention in Bihar and Jharkhand. Delayed ANC initiation may be significantly associated with unwanted pregnancy, but only when pregnancies were most decisively identified as unwanted. PMID- 19550161 TI - Individual- and contextual-level determinants of social inequities in under-five mortality in Nigeria: differentials by religious affiliation of the mother. AB - In sub-Saharan Africa, social factors such as religion have been linked with significant child morbidity and mortality. In religiously multi-denominational societies, one dominant religious group tends to subject the other groups to some form of economic, social or political discrimination, resulting in inequities in child survival. This study assessed the influence of individual- and community level indicators underlying the association between religion and under-five mortality using multilevel logistic regression modelling. There is a significant association between religion and under-five mortality, which is considerably attenuated by sequentially adjusting for other risk factors. The effect of religion on under-five mortality is influenced by contextual-level characteristics of the community. Traditional religion is associated with under five mortality, which is attributed to individual- and community-level indicators. Policy measures aimed at tackling child health inequities due to social factors such as religion need to consider the significance of the context in which mothers in different communities live, when healthcare interventions are being planned. PMID- 19550162 TI - Bacteriological assessment of stethoscopes used by medical students in Nigeria: implications for nosocomial infection control. AB - Our study assessed bacteria on swabs taken from the surface of the diaphragm of stethoscopes used by medical students in Nigeria. We found bacterial contamination on 80.1% of the stethoscopes. Staphylococcus aureus and Pseudomonas aeruginosa were major isolates. Bacterial colonization was highest among stethoscopes cleaned with only water and those never cleaned with any agent or never cleaned at all. The difference was statistically significant (chi2 = 31.9, p < .05). Stethoscopes from students who cleaned them after use on each patient and from those who practised handwashing after contact with each patient had significantly lower bacterial contamination (chi2 = 26.9; p < .05 and chi2=31.9, p < 0.05, respectively). Isolates of Staphylococcus aureus showed the highest susceptibility to antibiotics, while the most effective antibiotics were Ciproflox and Erythromycin. Integration of stethoscope care in the training curriculum of medical schools will enhance the control nosocomial infections. PMID- 19550163 TI - Re-positioning the role of traditional, complementary and alternative medicine as essential health knowledge in global health: do they still have a role to play? AB - Traditional, complementary and alternative medicine (TCAM), drawn from indigenous medical and/or healing knowledge systems from around the world, has for the last 30 years been recognized by the World Health Organization (WHO) as providing culturally acceptable, affordable and sustainable primary healthcare. TCAM knowledge has been known for some time to assist with birthing practices, acute injuries, infectious diseases and parasites. Although the focus on TCAM began in earnest by the WHO in 1978, and was re-emphasized between 2002 and 2008, TCAM has for the most part been overlooked in large-scale international health programs. This paper follows recent global interest in TCAM and examines notable developments that have specific relevance for TCAM integration in global primary healthcare. Drawing on established work by Bodeker and others, we focus on how TCAM is used in the context of health promotion, disease prevention and the reduction of infectious diseases. Specific examples include the use of TCAM practitioners for HIV/AIDS prevention awareness and direct treatment of AIDS related symptoms; the use of TCAM herbs for the treatment of malaria and the use of home herbal gardens for health maintenance. The final contribution of the paper helps to theorize inherent challenges and possible solutions to integrating TCAM into global health that have not been widely discussed to date. PMID- 19550164 TI - How to win an argument about the senior year of medical school. PMID- 19550165 TI - Becoming a woman physician scientist. PMID- 19550166 TI - Time for physicians to take action on climate change. PMID- 19550167 TI - Commentary: Informatics in biomedicine and health care. AB - During the last two decades, biomedical informatics (BMI) has become a critical component in biomedical research and health care delivery, as evidenced by two recent phenomena. One, as discussed in the article by Bernstam and colleagues in this issue, has been the introduction of Clinical and Translational Science Awards. Perhaps even more important has been the recent, arguably long overdue, emphasis on deployment of health information technology (IT) nationally. BMI utilizes IT and computer science as tools and methods for improving data acquisition, data management, data analysis, and knowledge generation, but it is driven by a focus on applications based in deep understanding of the science and practice, problems, interactions, culture, and milieu of biomedicine and health. Building from Bernstam and colleagues' distinction between BMI and other IT disciplines, the authors discuss the evolving role of BMI professionals as individuals uniquely positioned to work within the human and organizational context and culture in which the IT is being applied. The focus is not on the IT but on the combination--the interactions of IT systems, human beings, and organizations aimed at achieving a particular purpose. There has never been a time when the need for individuals well trained in BMI--those who understand the complexities of the human, social, and organizational milieu of biomedicine and health--has been more critical than it is now, as the nation seeks to develop a national infrastructure for biomedicine and health care, and as these fields seek to broadly deploy IT wisely and appropriately. PMID- 19550168 TI - Longitudinal, integrated clerkship education: better for learners and patients. Point. PMID- 19550169 TI - Longitudinal, integrated clerkship education: is different better? Counterpoint. PMID- 19550170 TI - What training is needed in the fourth year of medical school? Views of residency program directors. AB - PURPOSE: To identify common struggles of interns, determine residency program directors' (PDs') views of the competencies to be gained in the fourth year of medical school, and apply this information to formulate goals of curricular reform and student advising. METHOD: In 2007, semistructured interviews were conducted with 30 PDs in the 10 most common specialty choices of students at the University of California, San Francisco, School of Medicine to assess the PDs' priorities for knowledge, skills, and attitudes to be acquired in the fourth year. Interviews were coded to identify major themes. RESULTS: Common struggles of interns were lack of self-reflection and improvement, poor organizational skills, underdeveloped professionalism, and lack of medical knowledge. The Accreditation Council for Graduate Medical Education competencies of patient care, practice-based learning and improvement, interpersonal and communication skills, and professionalism were deemed fundamental to fourth-year students' development. Rotations recommended across specialties were a subinternship in a student's future field and in internal medicine (IM), rotations in an IM subspecialty, critical care, and emergency and ambulatory medicine. PDs encouraged minimizing additional time spent in the student's future field. Suggested coursework included an intensively coached transitional subinternship and courses to improve students' medical knowledge. CONCLUSIONS: PDs deemed the fourth year to have a critical role in the curriculum. There was consensus about expected fourth-year competencies and the common clinical experiences that best prepare students for residency training. These findings support using the fourth year to transition students to graduate medical training and highlight areas for curricular innovation. PMID- 19550171 TI - Artist's statement: my cadaver. PMID- 19550172 TI - Reflecting on reflections: enhancement of medical education curriculum with structured field notes and guided feedback. AB - The promotion of reflective capacity within the teaching of clinical skills and professionalism is posited as fostering the development of competent health practitioners. An innovative approach combines structured reflective writing by medical students and individualized faculty feedback to those students to augment instruction on reflective practice. A course for preclinical students at the Warren Alpert Medical School of Brown University, entitled "Doctoring," combined reflective writing assignments (field notes) with instruction in clinical skills and professionalism and early clinical exposure in a small-group format. Students generated multiple e-mail field notes in response to structured questions on course topics. Individualized feedback from a physician-behavioral scientist dyad supported the students' reflective process by fostering critical-thinking skills, highlighting appreciation of the affective domain, and providing concrete recommendations. The development and implementation of this innovation are presented, as is an analysis of the written evaluative comments of students taking the Doctoring course. Theoretical and clinical rationales for features of the innovation and supporting evidence of their effectiveness are presented. Qualitative analyses of students' evaluations yielded four themes of beneficial contributions to their learning experience: promoting deeper and more purposeful reflection, the value of (interdisciplinary) feedback, the enhancement of group process, and personal and professional development. Evaluation of the innovation was the fifth theme; some limitations are described, and suggestions for improvement are provided. Issues of the quality of the educational paradigm, generalizability, and sustainability are addressed. PMID- 19550173 TI - A program to enhance competence in clinical transaction skills. AB - The ability to take a comprehensive and accurate clinical history, perform a thorough and nuanced physical examination, engage in sequential clinical reasoning using all relevant clinical and laboratory data, and communicate clearly and compassionately with patients and other providers--the skills of the clinical transaction--are critical to a successful therapeutic outcome. Yet few medical schools' curricula include an explicit focus on developing these skills beyond the introductory level. Vanderbilt Medical School has developed a structured curriculum, integrated into the traditional clerkships of the third and fourth years, that ensures that each student receives specific instruction in clinical transaction skills. The clinical transaction curriculum is based on a set of 25 presenting problems, with learning objectives identified for each problem. Primary responsibility for instruction relating to each presenting problem is assigned to specific core clerkships, with the major portion of teaching provided by a nucleus of specially selected and compensated master clinical teachers. The Clinical Transaction Project at Vanderbilt was begun in 2004. Future development will focus on enhancing approaches to student assessment. PMID- 19550174 TI - Learning through longitudinal patient care-narratives from the Harvard Medical School-Cambridge Integrated Clerkship. AB - PURPOSE: Most medical schools value and seek to create opportunities for students to learn through experiences in the longitudinal care of patients. A number of innovative programs have made longitudinal care the central experiential component of principal clinical year education.The authors sought to identify ways in which learning through the longitudinal care of patients in an innovative longitudinal integrated clerkship contributes to the education of students in their principal clinical year. METHOD: The authors reviewed 16 narratives written by 14 of the 38 students from the first four years of the Harvard Medical School Cambridge Integrated Clerkship, 2004-2007, to identify important aspects of learning from longitudinal care. RESULTS: Students reported that the clerkship structure created a dynamic learning environment that helped them to more broadly learn about their patients' diseases and experiences of illness. Students described feeling deeply connected to "their" patients, which transformed their roles and inspired their reflections. With more thorough knowledge of their patients over time, they felt they made important contributions to their patients' care, not only in providing emotional support but also in bridging gaps in the delivery of services and in motivating deeper exploration into relevant medical and social issues. Students reported that their connections with patients over time inspired a sense of idealism and advocacy. CONCLUSIONS: Organizing learning in the principal clinical year around longitudinal patient care seems to offer significant advantages for learning and professional development. PMID- 19550175 TI - Teaching behavior change concepts and skills during the third-year medicine clerkship. AB - Risky health behaviors and social factors are linked to half of all causes of morbidity and mortality in the United States. Physicians report lack of training as one of the barriers to providing behavior change counseling. Formal behavior change curricula are infrequent in medical schools, and where they are available, they are often isolated from clinical experiences or presented through a limited approach. The authors developed the Health Beliefs and Behavior (HBB) course at University of Medicine and Dentistry-New Jersey Medical School (UMDNJ-NJMS) to teach the impact of unhealthy behaviors on health and wellness, to broaden students' understanding of the many factors that affect behavior, and to give medical students tools to facilitate health behavior change in patients. To the authors' knowledge, this is the only comprehensive, clinically integrated course on health behavior change in a U.S. medical school.The authors intercalated the 60-hour HBB course in the four-week, third-year internal medicine clerkship ambulatory block. Thus, students practice learned techniques in both the ambulatory and classroom settings, and they gain insight into health behavior by applying learned health models to patients and engaging in experiential exercises. Course components stress the biopsychosocial and patient-centered approach. The authors measure the impact of the course through student surveys. Third-year medical students at UMDNJ-NJMS who have completed the HBB course report enhanced understanding of the principles of behavior change and improved ability to perform behavior change counseling. PMID- 19550177 TI - Not so routine follow-up. PMID- 19550176 TI - The effect of gender on the clinical clerkship experiences of female medical students: results from a qualitative study. AB - PURPOSE: To characterize how female medical students perceive the role of gender within their medical education during the transition to the clinical curriculum. METHOD: In 2006-2007, the authors conducted a qualitative study consisting of in depth interviews with 12 third-year female medical students completing their first clinical clerkship. Participants were purposefully selected from a single New England medical school to represent a range of ages, ethnicities, and prior life experiences. RESULTS: Participants (1) struggled to define their role on the wards and often defaulted to stereotypical gender roles, (2) perceived differences in the nature of their workplace relationships compared with the nature of male medical students' workplace relationships, (3) had gendered expectations of male and female physicians that shaped their interactions with clinical supervisors, (4) felt able to negotiate uncomfortable situations with patients but felt unable to negotiate uncomfortable situations with supervisors and attendings, and (5) encountered a "gender learning curve" on the wards that began to shape their self-view as future female physicians. CONCLUSIONS: Despite increased numbers of women in medicine, issues of gender continue to have a substantial impact on the medical education of female students. Institutions can design interventions about gender issues in medicine that expand beyond a focus on sexual harassment to address the complex ways in which students are affected by issues of gender. PMID- 19550178 TI - Clerkship directors' practices with respect to preparing students for and using the National Board of Medical Examiners Subject Exam in medicine: results of a United States and Canadian Survey. AB - PURPOSE: Clerkship directors' practices regarding the National Board of Medical Examiners (NBME) subject exam in medicine are important in enhancing educational evaluation policy. The study's purpose was to determine clerkship directors' use of the subject exam in medicine and related learning activities in the context of curricula and outcomes of the directors' internal medicine clerkships. METHOD: The authors conducted a survey of directors of internal medicine clerkships in 2007. They performed descriptive statistical and multivariate analyses on all responses. RESULTS: Of 110 clerkship directors, 82 responded to the survey, for an overall response rate of 75%. Eighty-eight percent of the clerkship directors required the NBME subject examination in medicine. The mean minimum passing score was 62 (SD = 4.2); this score was not adjusted throughout the academic year, and it contributed 20% to 25% of the final grade. Most (89%) clerkships allowed students a retake after a failed first attempt. Most clerkship directors prepared students for the NBME subject exam in their programs through some combination of lectures, independent self-study, and review sessions with exam-preparation review books. However, 42% of clerkship directors lacked a specific strategy for a retake after a failure. CONCLUSION: Clerkship directors' use of the NBME subject exam in medicine is high. Most allow a retake after a first failure, and a combination of strategies is currently provided to help students prepare. A need exists to develop remediation plans for students who fail the exam. This report may serve as a reference for curricular and programmatic clerkship decisions. PMID- 19550179 TI - Transitional clerkship: an experiential course based on workplace learning theory. AB - Starting clerkships is anxiety provoking for medical students. To ease the transition from preclerkship to clerkship curricula, schools offer classroom based courses which may not be the best model for preparing learners. Drawing from workplace learning theory, the authors developed a seven-day transitional clerkship (TC) in 2007 at the University of California, San Francisco School of Medicine in which students spent half of the course in the hospital, learning routines and logistics of the wards along with their roles and responsibilities as members of ward teams. Twice, they admitted and followed a patient into the next day as part of a shadow team that had no patient-care responsibilities. Dedicated preceptors gave feedback on oral presentations and patient write-ups. Satisfaction with the TC was higher than with the previous year's classroom-based course. TC students felt clearer about their roles and more confident in their abilities as third-year students compared with previous students. TC students continued to rate the transitional course highly after their first clinical rotation. Preceptors were enthusiastic about the course and expressed willingness to commit to future TC preceptorships. The transitional course models an approach to translating workplace learning theory into practice and demonstrates improved satisfaction, better understanding of roles, and increased confidence among new third-year students. PMID- 19550181 TI - When the evaluated becomes evaluator: what can we learn from students' experiences during clerkships? AB - PURPOSE: To identify aspects that influence students' evaluation of the overall quality of clerkships and learning in clinical settings. METHOD: The authors analyzed 2,450 questionnaires dated 1997 through 2005 that evaluated clerkships of seven medical specialties (internal medicine, surgery, pediatrics, psychiatry, community medicine, emergency medicine, and obstetrics-gynecology). Students rated 22 questionnaire items addressing clerkships' global evaluation and domains related to structure, supervision, and clinical and problem-solving learning (PSL) activities using a five-point Likert scale. The authors performed statistical analysis using principal component analysis and regression analysis of items associated with students' global evaluation of clerkships. RESULTS: Correlation between clerkships' global ratings and ratings derived from the evaluation questionnaire was 0.871 (P < .0001). Clerkships' quality was mainly related to their organization, students' integration into clerkship, improvement of clinical skills, supervision, and residents' availability (r = 0.405; P < .0001). Among learning activities, opportunities for clinical practice predominated as the contributing factor to the overall perceived quality of most clerkships, but less than PSL activities in psychiatry (r = 0.070 versus 0.261, respectively; P < .001) and community medicine (r = 0.126 versus 0.298, respectively; P < .001); in surgery, both clinical practice and PSL activities contributed minimally to the clerkships' perceived quality (r = 0.150 and 0.148, respectively; P > .05). CONCLUSIONS: Factors influencing students' evaluation of a clerkship vary among medical specialties and depend not only on the teaching and teacher but also on the clerkship's organization, supervision, and learning activities. For clerkships where direct and multiple access to patients is more difficult, written case-based PSL activities proved complementary to direct patient encounter activities. PMID- 19550182 TI - The integrated clerkship: an innovative model for delivering clinical education at the Zhejiang University School of Medicine. AB - The traditional curriculum of clinical science at the Zhejiang University School of Medicine (ZUSM) was dominated by lectures, and many critical factors in producing competent physicians (such as the development of skills and active learning) were largely neglected. During a four-year period ending in 2007, ZUSM developed and implemented a new model for delivering clinical science education the integrated clerkship. The principal features of the model are the greater amount of time that students are exposed to and are working in the clinical environment as clerks and the great reduction in lecture hours. Unlike the U.S. model of clerkship, the integrated clerkship at ZUSM is characterized by a progressive process, with intensive preparatory lectures before the clerkship, which is divided into two levels, junior and senior. The junior clerkship is equally divided into didactic activities and clinical practice; the senior clerkship requires students to become an essential part of the work taking place on the wards. A preliminary program evaluation showed that the fundamental goals of the integrated clerkship had been largely attained, especially the mastery of basic clinical skills and retention of medical knowledge. Surveys showed that most of the integrated clerkship students and faculty members were satisfied with the new curriculum; the students felt better prepared to cope with the professional challenges of patient care, and they began to understand how social context affects their patients. As the pilot program in China, the integrated clerkship at ZUSM may serve as a template for medical schools at a similar level, in China and elsewhere. PMID- 19550184 TI - Longitudinal integrated clerkships for medical students: an innovation adopted by medical schools in Australia, Canada, South Africa, and the United States. AB - PURPOSE: Integrated clinical clerkships represent a relatively new and innovative approach to medical education that uses continuity as an organizing principle, thus increasing patient-centeredness and learner-centeredness. Medical schools are offering longitudinal integrated clinical clerkships in increasing numbers. This report collates the experiences of medical schools that use longitudinal integrated clerkships for medical student education in order to establish a clearer characterization of these experiences and summarize outcome data, when possible. METHOD: The authors sent an e-mail survey with open text responses to 17 medical schools with known longitudinal integrated clerkships. RESULTS: Sixteen schools in four countries on three continents responded to the survey. Fifteen institutions have active longitudinal integrated clerkships in place. Two programs began before 1995, but the others are newer. More than 2,700 students completed longitudinal integrated clerkships in these schools. The median clerkship length is 40 weeks, and in 15 of the schools, the core clinical content was in medicine, surgery, pediatrics, and obstetrics-gynecology. Eleven schools reported supportive student responses to the programs. No differences were noted in nationally normed exam scores between program participants and those in the traditional clerkships. Limited outcomes data suggest that students who participate in these programs are more likely to enter primary care careers. CONCLUSIONS: This study documents the increasing use of longitudinal integrated clerkships and provides initial insights for institutions that may wish to develop similar clinical programs. Further study will be needed to assess the long-term impact of these programs on medical education and workforce initiatives. PMID- 19550183 TI - Internal medicine clerkship characteristics associated with enhanced student examination performance. AB - PURPOSE: To determine which internal medicine (IM) clerkship characteristics are associated with better student examination performance. METHOD: The authors collected data from 17 U.S. medical schools (1,817 students) regarding characteristics of their IM clerkships, including structural characteristics, pedagogical approaches, patient contact, and clinical teacher characteristics. Outcomes of interest were postclerkship National Board of Medical Examiners (NBME) subject examination score, United States Medical Licensing Examination (USMLE) 2 score, and change in score from USMLE 1 to 2. To examine how associations of various clerkship characteristics and examination performance may differ for students of different prior achievement, the authors categorized students into those who scored in the top (1/4) of the cohort on USMLE 1 and the bottom (1/4). The authors conducted analyses at both the school and the individual student levels. RESULTS: In school-level analyses (using a reduced four-variable model), independent variables associated with higher NBME subject examination score were more small-group hours/week and use of community-based preceptors. Greater score increase from USMLE 1 to 2 was associated with students caring for more patients/day. Several variables were associated with enhanced student examination performance at the student level. The most consistent finding was that more patients cared for per day was associated with higher examination performance. More structured learning activities were associated with higher examination scores for students with lower baseline USMLE 1 achievement. CONCLUSION: Certain clerkship characteristics are associated with better student examination performance, the most salient being caring for more patients per day. PMID- 19550186 TI - Medicine and the arts. Ethics [excerpt] by Dietrich Bonhoeffer. Commentary. PMID- 19550189 TI - More old people classes. PMID- 19550188 TI - Who wants feedback? An investigation of the variables influencing residents' feedback-seeking behavior in relation to night shifts. AB - PURPOSE: The literature on feedback in clinical medical education has predominantly treated trainees as passive recipients. Past research has focused on how clinical supervisors can use feedback to improve a trainee's performance. On the basis of research in social and organizational psychology, the authors reconceptualized residents as active seekers of feedback. They investigated what individual and situational variables influence residents' feedback-seeking behavior on night shifts. METHOD: Early in 2008, the authors sent obstetrics gynecology residents in the Netherlands--both those in their first two years of graduate training and those gaining experience between undergraduate and graduate training--a questionnaire that assessed four predictor variables (learning and performance goal orientation, and instrumental and supportive leadership), two mediator variables (perceived feedback benefits and costs), and two outcome variables (frequency of feedback inquiry and monitoring). They used structural equation modeling software to test a hypothesized model of relationships between variables. RESULTS: The response rate was 76.5%. Results showed that residents who perceive more feedback benefits report a higher frequency of feedback inquiry and monitoring. More perceived feedback costs result mainly in more feedback monitoring. Residents with a higher learning goal orientation perceive more feedback benefits and fewer costs. Residents with a higher performance goal orientation perceive more feedback costs. Supportive physicians lead residents to perceive more feedback benefits and fewer costs. CONCLUSIONS: This study showed that some residents actively seek feedback. Residents' feedback-seeking behavior partially depends on attending physicians' supervisory style. Residents' goal orientations influence their perceptions of the benefits and costs of feedback seeking. PMID- 19550190 TI - A new measure of the cognitive, metacognitive, and experiential aspects of residents' learning. AB - PURPOSE: Psychometric data are presented for the Cognitive Behavior Survey: Residency Level (rCBS), a survey that profiles cognitive, metacognitive, and experiential aspects of residents' learning. METHOD: The authors asked 963 residents from seven medicine residencies of large academic medical centers to participate in their study and gathered data from the respondents during a three year period, 2000-2002. A factor analysis cross-validation design guided the development of rCBS's seven scales: memorization, conceptualization, reflection, independent learning, critical thinking, meaningful learning experience, and attitude toward educational experience. Interscale correlations and MANOVA provided preliminary evidence of scale construct validity. RESULTS: A total of 424 residents (44%) responded. With several minor exceptions, items for each scale loaded .40 or higher. Memorization did not correlate with any other scale, except correlating negatively with critical thinking. Higher-order thinking scales (conceptualization, reflection, independent learning, critical thinking) correlated with one another and with meaningful learning experience and attitude toward educational experience. The one exception: conceptualization did not correlate with critical thinking. MANOVA results reveal that residents who scored in the top 20% on the reflection scale conceptualized, learned independently, and thought critically more than did the bottom 20%. CONCLUSIONS: Results provide preliminary support for scale reliability and construct validity. As residencies seek to meet expectations of the Accreditation Council for Graduate Medical Education's Outcome Project, rCBS could prove useful in program evaluation, residents' self-assessment, and assessment by serving as a means to explore how residents learn, how residency programs affect learning behavior, and how clinically strong and weak residents differ in learning behaviors. PMID- 19550191 TI - The Educational Kanban: promoting effective self-directed adult learning in medical education. AB - The author reviews the many forces that have driven contemporary medical education approaches to evaluation and places them in an adult learning theory context. After noting their strengths and limitations, the author looks to lessons learned from manufacturing on both efficacy and efficiency and explores how these can be applied to the process of trainee assessment in medical education.Building on this, the author describes the rationale for and development of the Educational Kanban (EK) at Children's Hospital Boston- specifically, how it was designed to integrate adult learning theory, Japanese manufacturing models, and educator observations into a unique form of teacher student collaboration that allows for continuous improvement. It is a formative tool, built on the Accreditation Council for Graduate Medical Education's six core competencies, that guides educational efforts to optimize teaching and learning, promotes adult learner responsibility and efficacy, and takes advantage of the labor-intensive clinical educational setting. The author discusses how this model, which will be implemented in July 2009, will lead to training that is highly individualized, optimizes faculty and student educational efforts, and ultimately conserves faculty resources. A model EK is provided for general reference.The EK represents a novel approach to adult learning that will enhance educational effectiveness and efficiency and complement existing evaluative models. Described here in a specific graduate medical setting, it can readily be adapted and integrated into a wide range of undergraduate and graduate clinical educational environments. PMID- 19550192 TI - Development and evaluation of a simulation-based pediatric emergency medicine curriculum. AB - PURPOSE: The infrequency of severe childhood illness limits opportunities for emergency medicine (EM) providers to learn from real-world experience. Simulation offers an evidence-based educational approach to develop and practice clinical skills. METHOD: This was a two-phase, randomized trial with a wait-list control condition. The development phase (2005-2006) involved systematic curriculum and rating checklist creation, producing a six-case, simulation-based curriculum linked to three evaluation cases.In the validation phase (2006-2007), the authors randomized 69 residents from two EM residencies to either an intervention group that received the curriculum one month before the first assessment of all participants or a wait-list control group that received the identical curriculum three months later. A final assessment of all residents followed one month after that. Two raters evaluated all residents. Primary outcome measures are percentages of items completed correctly. The authors assessed rater agreement using intraclass correlation (ICC) and compared group performance using mixed model analysis of variance. RESULTS: ICCs surpassed 0.78. The instructional intervention produced a statistically significant effect for two of three evaluation cases for the validation phase of the study, a case x occasion interaction. Training year was significantly associated with better performance. In a multivariate analysis, training year and session correlated with score, but study group did not. CONCLUSIONS: A one-day, simulation-based pediatric EM curriculum produced limited results. The evaluation approach is reasonable and reproducible for the population studied. Instructional dose strength and factors may have limited curriculum effectiveness. Focused, frequent, and effortful instructional interventions are necessary to achieve substantial performance improvements. PMID- 19550193 TI - Integration strategies for using virtual patients in clinical clerkships. AB - PURPOSE: To explore students' perceptions of virtual patient use in the clinical clerkship and develop a framework to evaluate effects of different integration strategies on students' satisfaction and perceptions of learning effectiveness with this innovation. METHOD: A prospective, multiinstitutional study was conducted at six schools' pediatric clerkships to assess the impact of integrating Web-based virtual patient cases on students' perceptions of their learning during 2004-2005 and 2005-2006. Integration strategies were designed to meet the needs of each school, and integration was scored for components of virtual patient use and elimination of other teaching methodologies. A student survey was developed, validated, and administered at the end of the clerkship to 611 students. Data were analyzed using confirmatory factor analysis and structural equation modeling. RESULTS: A total of 545 students (89%) completed the survey. Overall student satisfaction with the virtual patients was high; students reported that they were more effective than traditional methods. The structural model demonstrated that elimination of other teaching methodologies was directly associated with perceived effectiveness of the integration strategies. A higher use score had a significant negative effect on perceived integration, but a positive effect on perceived knowledge and skills gain. Students' positive perceptions of integration directly affected their satisfaction and perception of the effectiveness of their learning. CONCLUSIONS: Integration strategies balancing the use of virtual patients with elimination of some other requirements were significantly associated with students' satisfaction and their perceptions of improved knowledge and skills. PMID- 19550194 TI - Temporary tattoos to simulate skin disease: report and validation of a novel teaching tool. AB - PURPOSE: Dermatology is a visual specialty requiring examination and description of skin lesions and the development of analytic skills to establish a diagnosis. Student education in dermatology is challenged by several factors. Although 10% to 15% of a general practitioner's consultations are related to the skin, dermatology is often underrepresented in undergraduate medical curriculums. In addition, more serious lesions, such as malignant melanoma (MM), are promptly biopsied and may not be available for students' examination. The authors carried out this study to learn whether a novel educational tool, a temporary tattoo, could successfully simulate an MM. METHOD: Eighty-one dermatologists and 14 dermatology residents participated in this validity study of a tattoo applied to the arm of a standardized patient (SP) to simulate an MM. The study was conducted at the 82nd Annual Canadian Dermatology Association Conference held in June 2007 in Toronto, Canada. RESULTS: A correct diagnosis was made by 93.8% (76/81) of the dermatologists and 90.5% of the participants (86/95) overall. The tattoo was also evaluated as being very realistic on a five-point Likert scale. CONCLUSIONS: The validation of the tattoo shows potential for use in medical education, such as SP visits and examinations. This teaching tool can be used to simulate a variety of skin lesions, providing a way to visually examine a lesion on the skin of an SP, which would enhance the medical student's learning experience. PMID- 19550196 TI - Perspective: Simulation and transformational change: the paradox of expertise. AB - Simulation is widely seen as a space where procedural skills can be practiced in safety, free from the pressures and complexities of clinical care. Central to this approach is the notion of simplification, a stripping down of skills into their component parts. Yet the definition of simplicity is contestable, often determined by experts without reference to those they teach.The author uses the ha-ha, a hidden ditch around a large country house used by 18th-century English landscape gardeners to create an illusion that the house is surrounded by untamed nature, as a metaphor for the differing perspectives of expert and novice. The author proposes that this difference of perspective lies at the heart of many current problems with simulation and simulators.This article challenges the philosophy of simplification, arguing that procedural skills should not be divorced from their clinical context and that oversimplification of a complex process can interfere with deep understanding. The author draws on Meyer and Land's notions of threshold concepts and troublesome knowledge and on his own experience with patient-focused simulation to propose an alternative view of simulation, framing it as a safe space which can reflect the uncertainties of clinical practice and recreate the conditions of real-world learning. By reintroducing complexity and human unpredictability, simulation can provide a safe environment for assisting the transformational change that is essential to becoming a competent clinician. PMID- 19550197 TI - Students' views on the use of real patients and simulated patients in undergraduate medical education. AB - PURPOSE: To determine students' views about the strengths and weaknesses of real patient interactions as opposed to simulated patient (SP) interactions in the undergraduate medical curriculum in order to evaluate how their strengths can be optimally used and weaknesses remedied. METHOD: Five focus-group interviews were conducted among fourth- and fifth-year medical students at Maastricht University in 2007, using a preestablished interview guide. The interviews were recorded, transcribed, and analyzed using qualitative methods. RESULTS: In general, the 38 participants considered real patient encounters more instructive and more authentic than SP encounters. However, students identified several strengths of SP encounters compared with real patient encounters. For example, SP interactions were helpful in preparing students for real patient interactions (particularly with regard to communication skills and self-confidence), in the teaching of "intimate" physical examination skills, such as gynecological examination skills, and in giving constructive feedback on communication skills. In contrast to what we had anticipated, taking a time-out was considered easier in real patient interactions. CONCLUSIONS: Both real patient interactions and SP interactions are considered indispensable to undergraduate medical education. Each encounter has unique strengths and weaknesses from the perspectives of students. On the basis of strengths and weaknesses that were identified, suggestions were made for the use of real patients and SPs in undergraduate medical education. PMID- 19550199 TI - AM Last Page: medical education in the Netherlands. PMID- 19550200 TI - Cerebral blood flow: sleeping beauty awakened by exercise. PMID- 19550201 TI - Osteoarthritis after anterior cruciate ligament injury: the quadriceps question. PMID- 19550198 TI - Synergies and distinctions between computational disciplines in biomedical research: perspective from the Clinical andTranslational Science Award programs. AB - Clinical and translational research increasingly requires computation. Projects may involve multiple computationally oriented groups including information technology (IT) professionals, computer scientists, and biomedical informaticians. However, many biomedical researchers are not aware of the distinctions among these complementary groups, leading to confusion, delays, and suboptimal results. Although written from the perspective of Clinical and Translational Science Award (CTSA) programs within academic medical centers, this article addresses issues that extend beyond clinical and translational research. The authors describe the complementary but distinct roles of operational IT, research IT, computer science, and biomedical informatics using a clinical data warehouse as a running example. In general, IT professionals focus on technology. The authors distinguish between two types of IT groups within academic medical centers: central or administrative IT (supporting the administrative computing needs of large organizations) and research IT (supporting the computing needs of researchers). Computer scientists focus on general issues of computation such as designing faster computers or more efficient algorithms, rather than specific applications. In contrast, informaticians are concerned with data, information, and knowledge. Biomedical informaticians draw on a variety of tools, including but not limited to computers, to solve information problems in health care and biomedicine. The paper concludes with recommendations regarding administrative structures that can help to maximize the benefit of computation to biomedical research within academic health centers. PMID- 19550203 TI - Regulatory mechanisms of cerebral blood flow during exercise: new concepts. AB - The response of the cerebral vasculature to exercise is different from the other peripheral vasculature (e.g., muscle); the cerebral vasculature has a small vascular bed and is strongly regulated by cerebral autoregulation and the partial pressure of arterial carbon dioxide. This review focuses on the integrative mechanisms underlying the regulation of cerebral blood flow during exercise. PMID- 19550202 TI - Sex differences and mechanisms of task-specific muscle fatigue. AB - Women can be less fatigable than men because of sex-related differences within the neuromuscular system that impact physiological adjustments during a fatiguing task. However, the involved mechanism(s) for the sex difference is task specific. This review explores the novel hypothesis that variation of the task will alter the magnitude of the sex-difference in muscle fatigue and the contribution of involved mechanisms. PMID- 19550204 TI - It pays to have a spring in your step. AB - In humans, a large portion of the mechanical work required for walking comes from muscle-tendons crossing the ankle joint. Elastic energy storage and return in the Achilles tendon during each step enhance the efficiency of ankle muscle-tendon mechanical work far beyond what is possible for work performed by knee and hip joint muscle-tendons. PMID- 19550205 TI - High-intensity interval training to maximize cardiac benefits of exercise training? AB - We hypothesized that high-intensity aerobic interval training results in a greater beneficial adaptation of the heart compared with that observed after low to-moderate exercise intensity. This is supported by recent epidemiological, experimental, and clinical studies. Cellular and molecular mechanisms of myocardial adaptation to exercise training are discussed in this review. PMID- 19550206 TI - A neuromuscular mechanism of posttraumatic osteoarthritis associated with ACL injury. AB - Anterior cruciate ligament (ACL) injury leads to early-onset osteoarthritis. Quadriceps weakness is a consequence of ACL injury and is considered to result from arthrogenic inhibition (AMI). AMI is the neurological "shutdown" of muscles surrounding an injured joint, preventing full activation, reducing strength, and promoting atrophy. As quadriceps function is critical for energy absorption, its dysfunction may contribute to posttraumatic osteoarthritis. PMID- 19550207 TI - Editors report on the Journal of Lower Genital Tract Disease. PMID- 19550208 TI - Commentary on: Statement on HPV DNA test utilization. PMID- 19550209 TI - Statement on HPV DNA test utilization. PMID- 19550211 TI - p16 Improves interobserver agreement in diagnosis of anal intraepithelial neoplasia. AB - OBJECTIVES: Evaluation of anal intraepithelial neoplasia (AIN) is subjective. Previous studies have shown p16 and Ki-67 expressions to correlate with AIN grade. Biomarkers like p16 and Ki-67 may improve interobserver agreement. The objectives were (1) to determine the extent of interobserver agreement in evaluating AIN on routine hematoxylin and eosin (H&E) sections and (2) to test whether p16 and/or Ki-67 staining improve interobserver diagnostic agreement. MATERIALS AND METHODS: Seventy-seven anal specimens were retrieved. Sections were stained with monoclonal antibodies against p16 and Ki-67. Blind to the original diagnoses, 4 pathologists assessed H&E alone, p16 alone, Ki-67 alone, and all 3 simultaneously. Diagnoses were normal/reactive, AIN I/HPV, AIN II, and AIN III. Agreement was calculated using kappa and S statistics. RESULTS: Pathologists were board certified and had 2 to 25 years (mean = 13.6 years) of experience. Fair agreement was observed using H&E diagnosis alone (kappa = 0.38, S = 0.56). The p16 diagnostic evaluation demonstrated the highest agreement (kappa = 0.57, S = 0.73). Interobserver agreement for Ki-67 alone and for H&E/p16/Ki-67 combined were comparable to that of H&E alone (kappa = 0.4, S = 0.54 and kappa = 0.44, S = 0.62, respectively). When the pathologists' diagnoses for all diagnostic evaluations were compared with consensus diagnoses, the lowest average magnitude of disagreement was seen with Ki-67 alone, followed by p16 alone, H&E/p16/Ki-67 combined, and H&E alone. CONCLUSIONS: Interobserver agreement for diagnosis of AIN was fair when based solely on H&E preparation. p16 alone improved interobserver agreement and demonstrated superior agreement when compared with H&E, Ki-67, and H&E/p16/Ki-67 combined. PMID- 19550210 TI - The accuracy of colposcopic grading for detection of high-grade cervical intraepithelial neoplasia. AB - OBJECTIVE: To relate aspects of online colposcopic image assessment to the diagnosis of grades 2 and 3 cervical intraepithelial neoplasia (CIN 2+). METHODS: To simulate colposcopic assessment, we obtained digitized cervical images at enrollment after acetic acid application from 919 women referred for equivocal or minor cytologic abnormalities into the ASCUS-LSIL Triage Study. For each, 2 randomly assigned evaluators from a pool of 20 colposcopists assessed images using a standardized tool online. We calculated the accuracy of these assessments for predicting histologic CIN 2+ over the 2 years of study. For validation, a subset of online results was compared with same-day enrollment colposcopic assessments. RESULTS: Identifying any acetowhite lesion in images yielded high sensitivity: 93% of women with CIN 2+ had at least 1 acetowhite lesion. However, 74% of women without CIN 2+ also had acetowhitening, regardless of human papillomavirus status. The sensitivity for CIN 2+ of an online colpophotographic assessment of high-grade disease was 39%. The sensitivity for CIN 2+ of a high grade diagnosis by Reid Index scoring was 30%, and individual Reid Index component scores had similar levels of sensitivity and specificity. The performance of online assessment was not meaningfully different from that of same day enrollment colposcopy, suggesting that these approaches have similar utility. CONCLUSIONS: Finding acetowhite lesions identifies women with CIN 2+, but using subtler colposcopic characteristics to grade lesions is insensitive. All acetowhite lesions should be assessed with biopsy to maximize sensitivity of colposcopic diagnosis with good specificity. PMID- 19550212 TI - Hear all about it: nightly television news coverage of cervical cancer vaccination in the United States. AB - OBJECTIVE: To examine the content of human papillomavirus (HPV)-related vaccination information presented during nightly national television news broadcasts in the United States. MATERIALS AND METHODS: A retrospective content analysis of HPV vaccination coverage on 5 major nightly US television networks from 2002 to 2007. The Vanderbilt Television News Archive was searched for keywords "Gardasil," "cervical cancer vaccination," "human papillomavirus vaccine," and "HPV vaccination." Each television news broadcast was categorized as follows: segment length (in seconds), network (American Broadcasting Company, Columbia Broadcasting Company, National Broadcasting Company, Cable News Network, or Fox Broadcasting Company), year of broadcast (2002-2007), and (4) presentation type. Air dates were plotted on a timeline to depict trends and linkages to 5 seminal events surrounding the development, efficacy, and controversy regarding HPV vaccination. RESULTS: During the 6-year period, a total of 27 HPV-related vaccination news broadcasts aired. News broadcasts ranged from 10 to 250 seconds, lasting an average of close to 2 minutes (mean +/- SD, 127.0 +/- 66.1 seconds). Most broadcasts presented information pertaining to HPV and cervical cancer, information on vaccine labeling, impact of the vaccine, and raised issues or concerns about the vaccine. More than half (66.7%) of news broadcasts were directly related to 5 seminal events surrounding the development, efficacy, and controversy regarding HPV vaccination. CONCLUSION: All 5 networks included within the Vanderbilt Television News Archive aired HPV vaccination content, with National Broadcasting Company and Columbia Broadcasting Company broadcasting most of the news stories during this time period. As compared with other medical related information presented on national nightly television news during this time period, HPV vaccination received a modest amount of coverage. PMID- 19550213 TI - Prevalence and risk factors for human papillomavirus and cervical intraepithelial neoplasia among HIV-positive women at a tertiary level hospital in India. AB - OBJECTIVES: The hypothesis to be tested was that the prevalence of human papillomavirus (HPV) and cervical intraepithelial neoplasia would be significantly higher in HIV seropositive women as compared with seronegative controls. Secondary aims were to determine the risk factors for HPV and cervical intraepithelial neoplasia and the HPV types in HIV-positive women. MATERIALS AND METHODS: A cross-sectional study of women 18 to 49 years old was done. Seventy five women who were HIV seropositive and 58 seronegative women, of whom 27 had HIV-positive partners, participated in the study. A Pap smear and a cervical swab for HPV were done. Women with Pap smear abnormality underwent colposcopy and large loop excision procedures if indicated. RESULTS: Ten (13.3%) HIV-positive women had high-grade squamous intraepithelial lesion as compared with 2 (3.4%) seronegative women (odds ratio [OR] 4.3; 95% CI = 0.9-41.7; p =.048). Among the HIV-positive women, 28 (37.3%) had high-risk HPV, whereas only 9 (15.5%) had high risk HPV among seronegative women (OR 3.2; 95% CI = 1.3-8.3; p =.009). Among women who were positive for high-risk HPV, the HIV-positive women were significantly more likely to have more than 1 HPV type (OR 7.4; 95% CI = 1.4 43.7; p =.005). Women who had coitus at less than 18 years of age were more likely to have high-risk HPV infection (OR 2.9; 95% CI = 1.2-6.2; p =.013) even after controlling for HIV status. CONCLUSIONS: HIV-positive women have a higher risk for multiple HPV infections as compared with seronegative women. Behavioral factors dominate HIV in determining HPV infections and resultant cervical neoplasia. PMID- 19550214 TI - Cytological follow-up of women older than 50 years with high-grade cervical smear treated by large loop excision. AB - OBJECTIVE: To evaluate cytological surveillance for women older than 50 years, to detect recurrent or residual disease after treatment of cervical intraepithelial neoplasia by loop excision. MATERIALS AND METHODS: Women undergoing a large loop excision for high-grade squamous intraepithelial lesion or glandular cytological abnormalities during a period of 4 years (2000-2003) were identified from the colposcopy database. Women younger than 50 years or with a history of previous loop excision were excluded. Clinical data, histology, and follow-up cytology results for up to 2 years after treatment were collected. RESULTS: Eighty-nine patients were identified. Age of the women ranged from 51 to 66 years, with a median of 51.5 years. Thirty-two (36%) had severe dyskaryosis, 53 (60%) had moderate dyskaryosis, and 4 (4%) had glandular abnormalities on cervical cytology before the loop biopsy. Cervical intraepithelial neoplasia (CIN) 2,3 and glandular abnormalities, CIN 1, and no abnormalities were found in 50 (56%), 18 (20%), and 19 (22%) loop specimens, respectively. Invasive disease was found in 2 (2%) cases. They were excluded from further analysis. The lesion was completely excised in 58 (65%) and incompletely excised in 23 (26%) patients. It was not possible to comment on the margin status in 8 (9%) cases. These were excluded from further analysis. Of the 23 women who had margins involved, 8 (35%) had ectocervical, 12 (52%) had endocervical, and 3 (13%) had both margins involved. All women had follow-up cervical smears at the cytology clinic. At 6-month follow up, 3 patients had persistent CIN and 4 had borderline changes on cervical smears. At 2 years follow-up, 3 patients had high-grade squamous intraepithelial lesion abnormalities, 2 of whom had clear margins at their loop biopsy earlier.Twenty percent of the women with positive endocervical margins on loop excision needed further treatment for residual or persistent disease on follow up. Overall, 4 (5%) of the 79 patients who had a loop biopsy went on to have cytological abnormalities suggestive of persistent/residual disease needing further treatment. CONCLUSION: Cytological surveillance for post-loop biopsy follow-up seems to be a good option for detecting residual disease in this high risk group of patients. PMID- 19550215 TI - Multidisciplinary colposcopy clinicopathology correlation meetings: an activity review. AB - OBJECTIVE: Multidisciplinary colposcopy clinicopathology correlation meetings are deemed to be an important aspect of colposcopic quality assurance and are often a focus of attention in colposcopy quality assurance peer-review assessments. Despite this, there are few data on such meetings detailing activity or providing benchmarks for audit. MATERIALS AND METHODS: A retrospective analysis of clinicopathology correlation meetings held during a 3-year period (2004-2006) at the University Hospital of North Staffordshire was performed. RESULTS: A total of 65 meetings were held on a 2 to 4 weekly basis. All meetings contained a representation from cytology, pathology, and colposcopy. A total of 518 cases were listed and 475 were discussed, representing 6.6% of the total patient attendances at the colposcopy clinic during the study period. The main indications for discussion were as follows: cytology/histology discrepancy (35%), cytology/colposcopy discrepancy (10%), management dilemma (25%), and invasive cancer review (18%). A small proportion of cases listed (8%) were not discussed because of administrative problems. Problems were encountered in the quality of documentation, inconsistencies in the recording of findings, conclusions, and management plans. CONCLUSIONS: Multidisciplinary colposcopy pathology meetings provide a valuable data resource for recording and analyzing challenging areas in the clinical management of women with abnormal cervical cytology. However, such meetings are time and labor intensive both in terms of personnel and preparation. National guidelines need to be developed to guide clinicians on the frequency and standards required from such meetings. PMID- 19550216 TI - Treatability by cryotherapy in a screen-and-treat strategy. AB - OBJECTIVES: We estimated the percentage of women infected with human papillomavirus (HPV+) who cannot be immediately treated with cryotherapy. MATERIALS AND METHODS: In a 10,000-woman Costa Rican cohort, we analyzed the 559 HPV+ women aged 25 to 55 years and estimated the proportion for whom immediate cryotherapy was not indicated (i.e., invasive cancer, large precancerous lesions, or benign abnormalities that risk failure such as large ectopy, squamocolumnar junction not visualized, polyps, ulcers, or distorted or atrophied cervix). To determine whether cryotherapy at time of baseline HPV screening would effectively treat HPV+ women, 2 expert gynecologists independently judged entire clinical histories (5-7 years of cytology, histology, and HPV tests) and a full longitudinal series of digitized cervical images. RESULTS: Reviewers judged 144 (25.8%) of 559 HPV+ women as not treatable by immediate cryotherapy. Among 72 women with cervical intraepithelial neoplasia grade 3 who would benefit most from a screening program, 35 (48.6%) were not treatable. In particular, 29 women (40.3%) were determined not treatable for reasons most likely associated with cryotherapy's inadequacy (lesion was large, suspected cancerous or in the endocervical canal or fornix). CONCLUSIONS: "Screen-and-treat" programs in low resource settings will soon use a rapid HPV test to screen older women once or twice in their lifetime, identifying women at higher risk for precancer. Our findings suggest that cryotherapy might not effectively treat many precancers, and other safe, low-technology treatment options could be required, in a scenario where all HPV+ women in this targeted group would receive cryotherapy at the same visit. PMID- 19550217 TI - The management of cervical intraepithelial neoplasia during pregnancy: is colposcopy necessary? AB - OBJECTIVE: The 2006 American Society for Colposcopy and Cervical Pathology Consensus guidelines state that it is acceptable to defer colposcopy until 6 weeks postpartum in pregnant patients with atypical squamous cells of undetermined significance (ASC-US) or low-grade squamous intraepithelial lesion (LSIL) cytology. Therefore, we sought to determine the incidence of cervical intraepithelial neoplasia (CIN) 2,3 in pregnant patients referred to a university colposcopy clinic. MATERIALS AND METHODS: A retrospective study identified all pregnant women with abnormal cytology referred to the University of Alabama at Birmingham colposcopy clinic between May 2005 and September 2007. After an institutional review board approval was obtained, demographic information, referral cytology, and histologic data were collected. The colposcopic impression was also obtained from the records. RESULTS: Six hundred twenty-five pregnant patients were identified. The mean age was 23 years (range, 14-44 years), the mean parity was 1 (range, 0-7), and the mean gestational age was 24 weeks (range, 4-39 weeks). The most common referral cytology was LSIL (41.0%), followed by ASC US (34.1%), and high-grade squamous intraepithelial lesion (13.6%). One hundred thirty-eight patients (22%) underwent cervical biopsy at the time of initial colposcopy. Forty-three patients had CIN 1, 28 patients had CIN 2, and 23 patients had CIN 3. Forty-four patients (32%) had no evidence of CIN on biopsy. There were no cases of invasive cervical cancer identified. Of the 469 patients with ASC-US and LSIL cytology, 20 of 78 patients who had a cervical biopsy were diagnosed with CIN 2,3. Of the 128 patients with high-grade intraepithelial lesion or high-grade squamous intraepithelial lesion cytology, 31 of 60 patients who had a cervical biopsy were diagnosed with CIN 2,3. Repeat colposcopy in the third trimester was performed on 47 patients. Only 3 of 13 patients with a repeat biopsy had CIN 2,3. CONCLUSIONS: Pregnant patients with ASC-US or LSIL cytology rarely have colposcopically suspected CIN 2,3 at their initial colposcopy that warrants a cervical biopsy; therefore, it is reasonable to defer the initial colposcopy in patients with ASC-US and LSIL until at least 6 weeks postpartum. PMID- 19550218 TI - Trichoepithelioma of the vulva: report of a case and review of the literature. AB - Trichoepithelioma is a rare benign skin lesion arising from hair follicles. It most commonly arises on the face and scalp. It is exceptionally rare on the vulva. We present a case and review the literature. PMID- 19550219 TI - Home study course: summer 2009. PMID- 19550221 TI - Vulvar cytology: useful or not? PMID- 19550222 TI - Femoral neck fractures: controversies and evidence. PMID- 19550223 TI - Perioperative considerations in geriatric patients with hip fracture: what is the evidence? AB - Geriatric hip fracture management requires a specialized treatment algorithm secondary to the complex medical and social needs of this patient demographic. The overall goal of the treatment is early mobilization, in an effort to prevent the complications associated with prolonged recumbency and to return the patient to functional activity. There is near-universal agreement among orthopedic surgeons that fractures about the hip require operative fixation, but surgical management in this patient population brings with it a set of issues that require important consideration. The current article reviews the perioperative considerations associated with geriatric hip fractures and takes an evidence based look at the complex issues involved in managing these patients. PMID- 19550224 TI - Internal fixation versus arthroplasty for displaced femoral neck fractures: what is the evidence? AB - A review of the current evidence for internal fixation versus hemiarthroplasty versus primary total hip arthroplasty for displaced femoral neck fractures was undertaken. At the meta-analysis level no difference in postoperative pain, function, or quality of life can yet be demonstrated. A significant difference in mortality has also not been found, but a trend towards higher mortality after primary arthroplasty is possible. Internal fixation (IF) has less morbidity, but a higher risk of revision and less cost-effectiveness. Independent adjudication for IF technique is rare in studies and bias towards higher revision rates due to technical failure is an issue. Randomized trials comparing IF with arthroplasty remain underpowered in specific subgroups of patients, in which IF revision rates could be acceptable. In hemiarthroplasty the data suggest minimal differences in outcome between the prosthesis types. The cementless Austin-Moore prosthesis is out-dated. Currently a cemented unipolar or bipolar, depending on costs, hemi arthroplasty is the treatment of choice for an elderly patient with functional limitations before the fracture. The role of modern, uncemented hemiarthroplasty designs are uncertain until more data are published. Total hip arthroplasty (THA) should be considered in any active older patient with a displaced femoral neck fracture. Patients with concomitant osteoarthritis, rheumatoid arthritis, or renal failure do poorly with other treatment options and should be treated with THA. Randomized trials have shown THA to be a cost-effective treatment with lower revision rates than IF. THA may also appear to be superior to hemiarthroplasty in specific subgroups, but larger trials are needed to confirm this observation. PMID- 19550226 TI - Interobserver reliability of classification systems to rate the quality of femoral neck fracture reduction. AB - OBJECTIVES: We compared the interrater reliability of five classification systems and surgeons' global impressions of the quality of femoral neck fracture reduction. METHODS: Six orthopedic trauma surgeons, six orthopedic nontrauma surgeons, and six orthopedic residents from three sites each rated 50 radiographs of postoperative femoral neck fractures fixated with cannulated screws or a sliding hip screw, using their overall impression, the Garden Index, Lowell's criteria, Lindquist and Tornkvist's criteria, the Western Infirmary Glasgow (WIG) angle, and standards established by a working group of orthopedic trauma surgeons. RESULTS: Reliability estimates for the Garden Index, Lowell's criteria, and the working group standards all fell within the range of moderate agreement [intraclass correlation coefficient (ICC) range 0.41-0.48], with no instrument achieving higher reliability than reviewers' overall impressions (ICC 0.49, 95% CI 0.39-0.61). Reviewers reached only fair agreement using Lindquist and Tornkvist's criteria and the WIG angle (kappa = 0.27 and 0.39, respectively). Trauma surgeons consistently achieved higher agreement than did nontrauma surgeons and trainees. CONCLUSIONS: Future studies using quality of reduction as an outcome measure or exploring the prognostic importance of reduction quality should measure this with trauma surgeons' overall impression, rather than with less experienced assessors or using one of the alternative instruments. PMID- 19550225 TI - Optimal internal fixation for femoral neck fractures: multiple screws or sliding hip screws? AB - INTRODUCTION: The number of hip fractures is likely to exceed 500,000 in the United States and 88,000 in Canada annually over the next 40 years. Hip fractures are associated with a 30% mortality rate at 1 year and profound temporary, and sometimes permanent, impairment of independence and quality of life. OBJECTIVES: Although much focus has centered around the comparison of arthroplasty versus internal fixation devices in the treatment of femoral neck fractures, the optimal approach for internal fixation has been largely ignored. Identifying the optimal technique for internal fixation could reduce the historically high rates of revision surgery that have fueled arguments against fixing patients with displaced femoral neck fractures. RESULTS: Both indirect and direct comparisons suggest a possible benefit for a sliding hip screw over multiple cancellous screws in reducing the need for revision surgery. The indirect nature of the comparison from the meta-analysis of arthroplasty versus internal fixation, and the small sample sizes, methodological limitations, and nonsignificant pooled estimate from the direct comparisons, leaves the issue very much in doubt. CONCLUSIONS: Although the rationale for arthroplasty continues to gain popularity, previous studies suggest that we have yet to identify the best approach for internal fixation. Previous trials suggest that the issue is largely unresolved and solutions will likely come from larger randomized trials comparing alternative devices for fixing the hip. PMID- 19550227 TI - Can we improve fixation and outcomes in the treatment of femoral neck fractures? The use of pharmaceuticals. AB - Femoral neck fracture rates are on the rise with an estimated 500,000 occurring annually by 2040. Despite the mainstay of open reduction and internal fixation, nonunion rates of up to 33% have been reported for displaced femoral neck fractures. Recently, increasing attention has been focused upon the role of osteobiologics to stimulate fracture repair. This article looks at the role of these anabolic compounds and discusses their potential future roles in augmenting the treatment of this condition. PMID- 19550229 TI - Unipolar versus bipolar hemiarthroplasty for femoral neck fractures: is there a difference? AB - Arthroplasty has been shown to reduce the rate of reoperation after femoral neck fracture. A number of studies have compared the outcomes of unipolar and bipolar arthroplasty. At 1- to 3-year follow-up, the functional outcomes of unipolar and bipolar arthroplasty are equivalent. Bipolar arthroplasty has a higher initial cost. However, the long-term results of acetabular wear and need for revision are unknown. PMID- 19550228 TI - Can we improve fixation and outcomes? Use of bone substitutes. AB - Hip fractures secondary to osteoporosis are common in the elderly. Stabilizing these fractures until union is achieved is a challenge due to poor bone stock and insufficient purchase of the implant to the bone. The reported high rate of complications has prompted extensive research in the development of fixation techniques. Furthermore, manipulation of both the local fracture environment in terms of application of growth factors, scaffolds, and mesenchymal cells and the systemic administration of agents promoting bone formation and bone strength has been considered as a treatment option with promising results. There are only a few evidence-based studies reporting on fixation augmentation techniques. This article reports on the efficacy of bone graft substitutes for the fixation of hip fractures, in particular calcium phosphates, which have been used as granules, cements, and implant coatings. PMID- 19550230 TI - Optimal arthroplasty for femoral neck fractures: is total hip arthroplasty the answer? AB - Although there is little doubt that arthroplasty of one form or another is appropriate treatment for many patients with a displaced femoral neck fracture, there is ongoing controversy about the relative merits of different types of arthroplasty among specific groups of patients. In particular, total hip arthroplasty is infrequently recommended because of concerns about the perceived high initial cost, increased risk of dislocation, and the anticipated low functional demands and life expectancy of the typical hip fracture patient. When viewed in this traditional context, the merits of total hip arthroplasty are not believed to outweigh the risks, except in the rare instance of a patient with preexisting symptomatic hip arthritis that sustains a hip fracture.Accumulating evidence suggests that a reevaluation of the role of total hip arthroplasty (THA) in patients with hip fracture is warranted. Recent comparative follow-up studies have documented superior and more durable function in a subset of patients with displaced femoral neck fractures after total hip replacement when compared to hemiarthroplasty or uncomplicated osteosynthesis. Economic analyses have suggested that the long-term cost of treatment favors total hip replacement because of the additional cost of treating failures of internal fixation and hemiarthroplasty in patients who survive 2 years or longer after their initial hip fracture. Finally, recent advances in the design of THA components such as the introduction of improved bearing surfaces allowing the use of larger femoral heads, combined with improved surgical techniques, may be making THA safer and less prone to dislocation and other mechanical complications.The purpose of this paper is to review the recent literature regarding the results of total hip replacement in patients with a displaced fracture of the femoral neck. Total hip replacement may have a larger role in the treatment of displaced femoral neck fractures than it has in the past. PMID- 19550231 TI - Selection of outcome measures for patients with hip fracture. AB - In designing a study protocol relating to hip fracture treatment and outcomes, it is important to select appropriate outcome instruments. Before beginning the process of instrument selection, investigators must gain a comprehensive understanding of the condition of interest and have a thorough knowledge of the expected benefits and harms of the proposed intervention. Adequate evidence of an intervention's effectiveness includes indication of impact on the patient's health. We provide a brief discussion about different ways that health and health measurement have been defined, including the International Classification of Function, Disability and Health (ICF), health-related quality of life (HRQOL), and cost-to-benefit analyses. We outline important properties (reliability, validity, sensitivity to change, and responsiveness) that a measurement instrument must demonstrate before being considered an acceptable means to measure outcome. Potential outcome measures relevant to patients with hip fracture are summarized, and important points to consider in the selection of outcome measures for a hypothetical research question in a hip fracture population are discussed. PMID- 19550233 TI - Minimally invasive hip fracture surgery: are outcomes better? AB - OBJECTIVES: Intertrochanteric hip fractures have high morbidity and mortality rates. The purpose of this study was to determine if minimally invasive plating, nailing, or external fixation operations lead to improved outcomes for intertrochanteric hip fractures compared with standard insertion of a sliding hip screw (SHS). DATA SOURCES: A systematic search of MEDLINE (1996 to June 2007) and EMBASE (1980 to June 2007) was performed. Results were limited to English language studies. References from eligible studies were reviewed to identify additional studies. STUDY SELECTION: Studies were selected for review based on the following criteria: prospective or retrospective studies comparing minimally invasive plating, nailing, or external fixation to standard insertion of an SHS, exclusion of intracapsular and subtrochanteric hip fractures, and report of outcome data by treatment group to allow for comparison. DATA EXTRACTION: The following outcomes were extracted from eligible studies: operative time, operative blood loss, intraoperative complications, postoperative drop in hemoglobin, postoperative pain, postoperative medical or fracture complications, wound complications, length of hospital stay, and post-fracture function. DATA SYNTHESIS: Sufficient data existed among 14 randomized controlled trials to perform a meta-analysis and calculate pooled relative risks for failure of fixation, blood transfusion, and mortality. Relative risks were calculated with 95% confidence intervals using a random-effects model, and an analysis of heterogeneity between pooled studies was conducted. Other outcome measures that were extracted from 17 comparative studies are reported as a systematic review. CONCLUSIONS: Although a significant heterogeneity exists between pooled studies, minimally invasive hip fracture plating, nailing, or external fixation was associated with a decrease in transfusion rate [relative risk of 0.63 as compared to standard SHS (95% confidence interval 0.41 to 0.96; I(2) = 83.6%)]. There was no significant difference for the other comparisons, including mortality between minimally invasive plating, nailing, or external fixation and standard insertion of an SHS. PMID- 19550232 TI - Treatment of displaced femoral neck fractures in the elderly: a cost-benefit analysis. AB - BACKGROUND: The optimal treatment for displaced femoral neck fractures in elderly patients is controversial. Compared with hemiarthroplasty (HA), internal fixation (IF) is associated with less operative trauma, bleeding, and possibly lower mortality at the expense of a higher reoperation rate and possibly increased cost. METHODS: We estimated the costs from a third party payer perspective after 1 year of 2 strategies (HA and IF) for the treatment of femoral neck fractures in patients over the age of 60 years. Using a decision board, we elicited patient preferences for the 2 operative approaches and calculated the net benefit using the willingness-to-pay technique. RESULTS: The 1-year projected cost of 1 IF was $18,100, and that of 1 HA was $15,843 (incremental cost of $2257 for each IF). Of 108 participants, 61 (56.5%) chose IF as the preferred treatment option and were willing to pay an average of $3.33 per month to have this option available if needed. In Ontario, the total incremental cost of performing IF in patients that choose it was $64,714,103, and the total societal benefit was $289,263,600, yielding a net benefit of $224,549,497. CONCLUSION: The benefits of IF over HA outweigh the incremental costs from the perspective of a third-party payer. IF should be available to patients that choose it. PMID- 19550234 TI - Overcoming barriers to osteoporosis care in vulnerable elderly patients with hip fractures. AB - Indications for and benefits of providing osteoporosis (OP) care for hip fracture patients have become widely understood. The hip fracture patient is frequently over age 80 years, minimally ambulatory, has multiple medical comorbidities, and has cognitive impairment. Patient barriers to initiation of effective OP treatment include: age, dementia, medical comorbidities, polypharmacy, lack of adherence with treatment, alcohol abuse, postoperative delirium, language barriers, inadequate social support, and socioeconomic status. In a large teaching hospital, 244 patients presented with hip fracture over 2 years: 72% were female and 64% were over age 80. Forty percent had been diagnosed with dementia; another 29% had other severe medical comorbidities.Opportunities for OP diagnosis and treatment are numerous. In acute care hospitals, coordinator facilitated programs are effective for identification, education, assessment, referral, and treatment of underlying OP in fracture patients. System modifications may include an automated care path or automatic specialist referral for hip fracture patients. In the rehabilitation hospital, the patients are in a more stable condition, there is a focus on the recent fracture, and there are opportunities to initiate OP treatment and to promote adherence. In long-term care, dietary intake including calcium and vitamin D supplementation and persistence with pharmacotherapy can be monitored. Patient education and referral to the family physician for osteoporosis investigation and treatment have improved patient knowledge and diagnosis, but the reported impact on treatment has been limited.Effective OP care for the vulnerable hip fracture patient should be initiated early but may be complex and require coordination. In addition to calcium and vitamin D supplementation, most patients in this category have an indication for aminobisphosphonate therapy. Liaison between the orthopaedic team and the discharge destination caregivers, an established discharge diagnosis of osteoporosis, and ensuring patients are discharged on supplements and medication will promote patient, caregiver, and primary care physician awareness of the patient's OP care needs. Education programs may provide benefits at later stages, to improve adherence with treatment. PMID- 19550235 TI - Gamma nails revisited: gamma nails versus compression hip screws in the management of intertrochanteric fractures of the hip: a meta-analysis. AB - BACKGROUND: Concerns about the Gamma nail have largely been fueled by early randomized trials and meta-analyses suggesting an increased risk of subsequent femoral shaft fractures when compared with compression hip screws. Whereas meta analyses favor compression hip screws over first-generation Gamma nails, little is known as to whether the newer Gamma nail designs and the improved learning curve associated with the implants have reduced the risk of femoral shaft fracture. The current meta-analysis aimed to explore the effects of time and Gamma Nail design on the risks of femoral shaft fracture after treatment of extracapsular hip fractures. METHODS: We searched computerized databases (Medline, Cochrane, and SciSearch) for published randomized clinical trials from 1969 to 2002, and we identified additional studies through hand searches of major orthopedic journals, bibliographies of major orthopedic texts, and personal files. Two investigators independently graded study quality and abstracted relevant data. We abstracted information on subsequent femoral shaft fracture rates from studies. We pooled data using a random-effects model and tested for heterogeneity using the I test. We conducted sensitivity analyses by date and by generation of the Gamma nail. We further conducted a cumulative meta-analysis to explore the treatment effect over time. RESULTS: We identified 25 relevant randomized trials from 1991 to 2005. In earlier studies (N = 1585 patients), Gamma nails increased the risk of femoral shaft fracture 4.5 times compared with a compression hip screw (95% confidence interval: 1.78-11.36, P = 0.0014, I(2) = 0%). However, among the most recent studies (2000-2005), Gamma nails did not significantly increase femoral shaft fracture risk (relative risk = 1.65, 95% confidence interval: 0.50-5.44, P = 0.41, I(2) = 0%). The most recent study (2005) found no difference in femoral fracture rates (relative risk = 1.03, 95% confidence interval = 0.06-16.2, P = 0.99). CONCLUSIONS: Our meta-analysis of randomized trials suggests that previous concerns about increased femoral shaft fracture risk with Gamma nails have been resolved with improved implant design and improved learning curves with the device. Earlier meta-analyses and randomized trials should be interpreted with caution in light of more recent evidence. PMID- 19550236 TI - Intramedullary versus extramedullary fixation for subtrochanteric femur fractures. AB - OBJECTIVES: Both intramedullary and extramedullary internal fixation has been advocated for the treatment of subtrochanteric femur fractures. Is there clinical evidence to recommend one method of internal fixation over the other? DATA SOURCES: A search of MEDLINE (1950 to June 2007), CINAHL (1982 to June 2007), and EMBASE (1980 to June 2007) was performed. Results were limited to English language studies. References from eligible studies were reviewed to identify additional studies. STUDY SELECTION: Studies were selected for review based on the following criteria: comparison and observational studies examining the use of intramedullary and/or extramedullary implants for the fixation of subtrochanteric femur fractures, inclusion of intertrochanteric hip fractures with subtrochanteric extension, exclusion of pure intertrochanteric and intracapsular hip fractures, and exclusion of pathologic fractures. DATA EXTRACTION: The following outcomes were extracted from eligible studies: operative time, operative blood loss, intraoperative complications, postoperative medical complications, number of patients transfused, wound complications, failure of fixation, rate of nonunion, length of hospital stay, and functional recovery. DATA SYNTHESIS: Three level I and 9 level IV studies were identified and used in the systematic review of outcomes for intramedullary and extramedullary fixation for subtrochanteric fractures. Three level I studies were used to calculate a pooled relative risk for failure of fixation with a 95% confidence interval. An analysis of heterogeneity between pooled studies was conducted. CONCLUSIONS: There is grade B evidence that operative time is reduced and that fixation failure is reduced with the use of intramedullary implants for subtrochanteric fractures. Future studies should perform subgroup analysis according to the type of population sampled (ie, young versus elderly) and subtrochanteric fracture type. PMID- 19550237 TI - Salvage of failed hip fracture fixation. AB - Nonunion of fractures about the femoral neck and intertrochanteric hip regions is uncommon. Patients who develop nonunions of these fractures typically exhibit marked pain and disability, thereby presenting a treatment challenge to the orthopaedic surgeon. Factors that guide the choice of salvage treatment include the anatomic site of the nonunion, the quality of the remaining proximal bone and articular surface, and patient factors (such as age and activity level). In the younger patients with a well-preserved hip joint, treatment typically involves revision internal fixation with or without osteotomy or bone grafting. However, in older patients, it is more common to encounter poor remaining proximal bone stock or a badly damaged hip joint from hardware cutout. As such, conversion to hip arthroplasty is intended to help effectively restore function and relieve pain. With respect to salvage procedures for the femoral head, the major challenges in decision making include the choice of both internal fixation device and accurate preoperative planning. The challenges involved in planning to convert to hip arthroplasty include the need for acetabular resurfacing, selecting the femoral implant, and managing discontinuity of the greater trochanter. Furthermore, there are additional technical challenges that may be encountered, such as broken hardware, deformity, and femoral bone defects. Overall, salvage of nonunions of femoral neck and intertrochanteric hip fractures in properly selected patients can provide patients with good to excellent results. PMID- 19550239 TI - So far, so good for lung cancer SBRT. PMID- 19550240 TI - "The best-laid plans ... often go awry ...". PMID- 19550241 TI - Survival prognostication in non-small cell lung cancer. PMID- 19550242 TI - Class IA phosphatidylinositol 3-kinase signaling in non-small cell lung cancer. PMID- 19550243 TI - Regression of established AB1 murine mesothelioma induced by peritumoral injections of CpG oligodeoxynucleotide either alone or in combination with poly(I:C) and CD40 ligand plasmid DNA. AB - INTRODUCTION: Stimulation of the CD40 receptor using an agonistic anti-CD40 antibody can slow the growth of AB1 tumors. Stimulation of the GITR receptor may also have antitumor activity by countering the immunosuppressive effects of regulatory CD4 T cells. Similarly, agonists for Toll-Like Receptors (TLR) such as CpG oligodeoxynucleotides (TLR9 agonist) have activity against AB1 tumors. Combinations of CpG with CD40 ligand and polyinosinic-polycytidylic acid (poly(I:C), TLR3 agonist) may be even stronger than CpG alone. The synergistic effects of these combinations have been tested in other tumor types but not in mesothelioma. METHODS: Established AB1 mesothelioma tumors were injected with either plasmid DNA encoding a novel 4-trimer form of murine CD40 ligand (pSP-D CD40L), GITR ligand (GITRL), or control plasmid DNA. In addition, CpG with or without poly(I:C) was also injected intratumorally. RESULTS: Plasmid injections of pSP-D-CD40L or pSP-D-GITRL, had no significant antitumor effect, possibly reflecting the difficulty of administering DNA injections into this very dense tissue. However, the injection of CpG with or without poly(I:C) strongly suppressed tumor growth and led to long-term tumor-free survival. The response to a triple combination of pSP-D-CD40L + CpG + poly(I:C) was demonstrated by an increase in intratumoral CD8 T cells and a dramatic increase in F4/80 macrophages. CONCLUSIONS: Intratumoral injections of plasmid DNAs encoding highly active forms of either CD40 ligand or GITR ligand had no significant antitumor effects in this model, although improved DNA delivery techniques could possibly improve this strategy. In contrast, intratumoral CpG injections had significant antitumor effects and there were indications that CpG plus poly(I:C) was even more effective. Taken together, these data confirm previous reports that immune stimulants, especially CpG TLR9 agonists, have potential as a treatment for mesothelioma. PMID- 19550244 TI - Differences in the quality of information on the internet about lung cancer between the United States and Japan. AB - INTRODUCTION: Quality of information available over the Internet has been a cause for concern. Our goal was to evaluate the quality of information available on lung cancer in the United States and Japan and assess the differences between the two. METHODS: We conducted a prospective, observational Web review by searching the word "lung cancer" in Japanese and English, using Google Japan (Google-J), Google United States (Google-U), and Yahoo Japan (Yahoo-J). The first 50 Web sites displayed were evaluated from the ethical perspective and for the validity of the information. The administrator of each Web site was also investigated. RESULTS: Ethical policies were generally well described in the Web sites displayed by Google-U but less well so in the sites displayed by Google-J and Yahoo-J. The differences in the validity of the information available was more striking, in that 80% of the Web sites generated by Google-U described the most appropriate treatment methods, whereas less than 50% of the Web sites displayed by Google-J and Yahoo-J recommended the standard therapy, and more than 10% advertised alternative therapy. Nonprofit organizations and public institutions were the primary Web site administrators in the United States, whereas commercial or personal Web sites were more frequent in Japan. CONCLUSION: Differences in the quality of information on lung cancer available over the Internet were apparent between Japan and the United States. The reasons for such differences might be tracked to the administrators of the Web sites. Nonprofit organizations and public institutions are the up-and-coming Web site administrators for relaying reliable medical information. PMID- 19550245 TI - A pericardiophrenic vein varix mimicking a pulmonary mass. PMID- 19550246 TI - Pleural well-differentiated papillary mesothelioma: a case report. AB - Well-differentiated papillary mesothelioma is a pathologic entity distinct from malignant mesothelioma and has a different tumor biology highlighted by its relatively good prognosis. We report the case of a 27-year-old woman with pleural well-differentiated papillary mesothelioma, who was treated with bilateral pleurodesis and pericardial window to control the production of malignant fluid, without significant systemic anticancer treatment. She survived for 16 years, confirming its indolent nature. This case report also highlights the role of fluid cytology, immunohistochemistry, and electron microscopy in the diagnosis of well-differentiated papillary mesothelioma and the potential use of serum CA-125 as a marker of progression. PMID- 19550247 TI - Invasive inflammatory myofibroblastic tumor of the lung. AB - Inflammatory myofibroblastic tumor (IMT) of the lung is a rare tumor but it should be considered when dealing with primary lung tumors in children, adolescents, and nonsmoking adults. It is, from a pathologic point of view, a benign tumor composed of a spindle cell proliferation and inflammatory cells. Its clinical behavior, however, is variable with a benign evolution at one, and a malignant evolution with recurrent and metastatic disease at the other end of the spectrum. Diagnosis is very difficult and often only possible after resection of the tumor. We present a case of pulmonary IMT in a 15-year-old male with malignant features on radiographic and F-Fluoro-deoxyglucose positron emission tomography imaging. Pathogenesis, pathology findings, clinical behavior, and imaging of pulmonary IMT are briefly discussed. PMID- 19550248 TI - Extraskeletal osteosarcoma arising in anterior mediastinum: brief report with a review of the literature. AB - Extraskeletal osteosarcoma arising from the mediastinum is an extremely rare malignant tumor. We present the case of a 77-year-old woman with a past history of pulmonary tuberculosis who presented with a dyspnea. Imaging studies revealed a huge anterior mediastinal mass with scattered calcifications. Complete surgical resection was successfully performed, and histologically the tumor contained osteoid formation consisting of round atypical cells. The literature concerning this rare tumor is reviewed. PMID- 19550250 TI - Epidermal growth factor receptor exon 19 deletions predict complete regression of multiple intracranial metastases in two cases of non-small cell lung cancer treated with erlotinib. PMID- 19550249 TI - Summary of selected presentations from the 8th annual targeted therapy in lung cancer symposium. AB - Lung cancer is the leading cause of cancer-related death in the United States. Outcomes for patients with lung cancer have reached a plateau with cytotoxic chemotherapy. Lung cancer remains very much at the vanguard of the new revolution in cancer therapy using molecular targets. Although striking improvements in survival have been observed in the treatment of chronic myeloid leukemia, gastrointestinal stromal tumors, and in a subset of breast cancer using this approach, the impact of targeted therapies in lung cancer is quite modest. Along with advances in imaging and cancer genomics, there is now considerable optimism that the pace of progress in the treatment of lung cancer will accelerate in the next 10 years. As has been the custom for the past 8 years, leading experts in the biology, diagnosis and treatment of lung cancer met for three days to discuss current areas of research and future directions. This summary provides a brief snapshot of the discussions held at the 8th Annual Meeting on Targeted Therapies for Lung Cancer sponsored by the International Association for the Study of Lung Cancer in Santa Monica in early February 2008. PMID- 19550251 TI - Asymptomatic air embolism after computed tomography-guided lung needle marking. PMID- 19550238 TI - Resolving controversies in hip fracture care: the need for large collaborative trials in hip fractures. AB - Hip fractures are a significant cause of morbidity and mortality worldwide and the burden of disability associated with hip fractures globally vindicate the need for high-quality research to advance the care of patients with hip fractures. Historically, large, multi-centre randomized controlled trials have been rare in the orthopaedic trauma literature. Similar to other medical specialties, orthopaedic research is currently undergoing a paradigm shift from single centre initiatives to larger collaborative groups. This is evident with the establishment of several collaborative groups in Canada, in the United States, and in Europe, which has proven that multi-centre trials can be extremely successful in orthopaedic trauma research.Despite ever increasing literature on the topic of his fractures, the optimal treatment of hip fractures remains unknown and controversial. To resolve this controversy large multi-national collaborative randomized controlled trials are required. In 2005, the International Hip Fracture Research Collaborative was officially established following funding from the Canadian Institute of Health Research International Opportunity Program with the mandate of resolving controversies in hip fracture management. This manuscript will describe the need, the information, the organization, and the accomplishments to date of the International Hip Fracture Research Collaborative. PMID- 19550252 TI - On our way. PMID- 19550253 TI - Is the new Joint Commission mandate that healthcare institutions address disruptive behaviors an appropriate standard? Pro position. PMID- 19550254 TI - Is the new Joint Commission mandate that healthcare institutions address disruptive behaviors an appropriate standard? Con position. PMID- 19550255 TI - Hypoxia in the term newborn: part two-primary pulmonary disease, obstruction, and extrinsic compression. AB - Pediatric care providers are repeatedly called upon to evaluate a cyanotic newborn in the labor and delivery suite, or in the well-baby nursery. A myriad of disorders spanning all-organ systems exist as possibilities for each of these problems, although several causes for newborn cyanosis are particularly common. In this second of a three-part series, primary pulmonary disease, airway obstruction, and extrinsic compression of the lungs as causes for newborn hypoxia are explored. It is in this group of disorders that we find the answers for the greatest number of these cyanotic dilemmas. Knowledge of the breadth of diagnoses, and respect for the variety of clinical possibilities, is the first step in providing a patient with accurate diagnosis, treatment, and referral. PMID- 19550257 TI - The experience of dyspnea in school-age children with asthma. AB - PURPOSE: To explore the experience of dyspnea in school-age children with asthma including exploring children's perceptions of the (1) sensations of dyspnea, (2) precipitants of dyspnea, (3) coping strategies used to deal with dyspnea, and (4) effects of dyspnea on lives of children. STUDY DESIGN AND METHODS: This interpretive, descriptive, qualitative research study had a sample of 30 school age children diagnosed with asthma. Data collection involved individual open ended interviews combined with drawings. Transcribed data were analyzed using the constant comparative method. RESULTS: The childrens' experiences with dyspnea were represented by five themes: (1) it is an overwhelming feeling, (2) it is mainly..., (3) I slow it down, (4) others only need to help when it is really bad, and (5) I am not a player. Although children varied with respect to how they described their experiences, they all reinforced that the sensation of dyspnea was distressing and painful, something that when experienced overshadowed everything else. CLINICAL IMPLICATIONS: Children with dyspnea have much to share about what it is like to experience dyspnea that may be used by nurses to provide comprehensive and sensitive care. Nurses need to take into account the individuality of children's dyspnea experiences when developing treatment plans for children with asthma. Education programs that are tailored to meet individual needs will help children to take control and manage their dyspnea. PMID- 19550259 TI - Out of the abyss of colic: a view through the fathers' eyes. AB - PURPOSE: To elucidate the experience of fathers living with a colicky infant. STUDY DESIGN AND METHODS: Interpretive phenomenology using in-depth interviews with 10 fathers of colicky infants. Reiterative interview reading, repeated meaning unit sorting, going back to the literature, and discussions gave way to emerging themes. FINDINGS: The overall experience is one of "falling into and arising from the crying abyss together as a family." Four themes emerged: (a) falling in, (b) hitting bottom, (c) weaving strands to make a rope, and (d) climbing out. CLINICAL IMPLICATIONS: From their stories, fathers provide insight useful to nurses assisting families of colicky infants through a very difficult time. Nurses should talk to fathers directly about the possibility of colic, and teach both parents what it is, how long it usually lasts, and what can be done about it. Anticipatory guidance as well as discussion about colic during newborn health visits can help parents to vocalize their feelings, and learn what actions to take when the stress of colic becomes overwhelming. PMID- 19550260 TI - Girls in foster care: a vulnerable and high-risk group. AB - PURPOSE: To describe the characteristics of girls in foster care who have exhibited sexually abusive behavior. STUDY DESIGN AND METHODS: This was a chart review and a descriptive, exploratory study of 155 female children and adolescents (age 4-17 years) who were in foster care and had been evaluated for their sexual abusive and aggressive behaviors. RESULTS: Findings indicated that these girls had significant histories of maltreatment, chronic health issues, and foster care placement instability; 84% of the girls had been physically abused; 95% were victims of ongoing abuse by the biological parents (78%). Sexual abuse was reported by 81%, and 68% were sexually abused by more than one individual. Ninety-five percent were neglected; 51% of the neglect was classified as severe and chronic. All of the girls (100%) had been shown to exhibit sexually abusive and inappropriate behaviors toward other children, including exposing themselves to age-mates, violation of body space, sexually aggressive remarks, sexual touching without permission, and sexual touching of much younger children. Ninety two percent of the girls had two or more changes in caregivers by age 16 years. Over one-third of the girls (39.2%) experienced four or more different living situations that lasted less than 1 month. CLINICAL IMPLICATIONS: These girls demonstrated a variety of behavior problems that place them at high risk for multiple foster care placements, which can negatively impact upon their well being and health. Whether in healthcare settings, schools, the workplace, community, in-patient, or psychiatric settings, nurses and other healthcare providers have access to children and their foster care caregivers. Nurses and other healthcare clinicians are in an ideal position to provide a safe clinical environment contributing to the health, education, and support to these girls. PMID- 19550261 TI - Infant hospital abduction: security measures to aid in prevention. AB - The purpose of this article is to describe measures used to prevent abduction of infants from healthcare facilities. According to the National Center for Missing & Exploited Children, 122 infants were abducted from hospitals between 1983 and 2008; characteristics of the abductors have been found to be similar in the majority of infant abductions. The Joint Commission has established security standards, and conducts assessments to identify abduction risk within hospitals. Nurses can contribute to the safety and security of infants by following prevention plans which emphasize these and other actions: educating parents about abduction risks, using identically numbered bands on the baby and parents, taking color photographs of the infant, wearing color photograph ID badges themselves, discouraging parents/families from publishing birth notices in the public media, controlling access to nursery/postpartum unit, and utilizing infant security tags or abduction alarm systems. PMID- 19550262 TI - Explanatory models of asthma in the southeast asian community. AB - PURPOSE: To examine how two groups (parents and healthcare providers) perceive asthma in Southeast (SE) Asian children in the United States, and to address issues of access to asthma care. STUDY DESIGN AND METHODS: Using Kleinman's Explanatory Models (EM) approach, semistructured interviews were used with 12 families and 26 providers. Families were interviewed in a clinic setting. Providers read a case example of a SE Asian child with asthma and were asked to anticipate the case family's EM of asthma. Data were analyzed using template analytic technique in which segments were clustered into etiology, causation, treatments, asthma impact, and access to asthma care. RESULTS: Families predominately followed current asthma practice guidelines, whereas providers believed that SE Asian families primarily followed cultural practices. Families described the severity and impact of asthma as more intense than the providers described the same items. While families identified barriers to care as difficulty getting appointments and unavailable providers, providers viewed family barriers to be predominately culturally based. Both groups noted the need for education and frequent healthcare visits as facilitators to care. CLINICAL IMPLICATIONS: This study suggests that important differences may exist between the way providers and SE Asian families perceive childhood asthma. Nurses are encouraged to ask the families in their care how they explain their children's asthma, thus facilitating more culturally competent care and increased ability to meet the family's needs. Implications for policy change relative to improving access to care to immigrant groups are also suggested. PMID- 19550263 TI - Infant nutrition: the challenge of colic in infants. PMID- 19550265 TI - Global health and nursing: overcoming poverty: one millennium village at a time. PMID- 19550264 TI - The new networking: family-initiated rapid response. PMID- 19550272 TI - The financial crisis and keeping patients safe. PMID- 19550273 TI - Should patients with a strong family history of pancreatic cancer be screened on a periodic basis for cancer of the pancreas? PMID- 19550274 TI - Natural course of acute pancreatitis: what we know today and what we ought to know for tomorrow. PMID- 19550276 TI - Tea consumption and the risk for pancreatic cancer. PMID- 19550277 TI - Th1 and Th2 profiles in patients with pancreatic cancer compared with chronic pancreatitis. PMID- 19550278 TI - Effect of low-molecular weight trypsin inhibitor, nafamostat mesilate, on trypsin activity using the pancreatic acinar cells. PMID- 19550279 TI - Percutaneous transcatheter thrombolysis for graft thrombosis after pancreas transplantation. PMID- 19550280 TI - Phenotype expression in a case of adult cystic fibrosis caused by an extremely rare compound heterozygous genotype (2183AA>G/2789+5G>A). PMID- 19550281 TI - Rapid changes in sclerosing cholangitis associated with autoimmune pancreatitis. PMID- 19550282 TI - Enhanced inhibition of tumor growth with depletion of CD25 regulatory cells and intratumoral immunization with tumor RNA-pulsed dendritic cells in a C57BL/6 pancreatic tumor model. PMID- 19550283 TI - The effect of HAART in 254 consecutive patients with AIDS-related Kaposi's sarcoma. AB - OBJECTIVE: A prospective cohort study was performed to evaluate the clinical outcomes of patients with histologically confirmed AIDS-related Kaposi's sarcoma diagnosed since the introduction of HAART. METHODS: Two hundred and fifty-four consecutive patients (96% men) diagnosed with Kaposi's sarcoma between 1996 and 2008 are included. Clinicopathological and treatment details were prospectively collected. The median follow-up is over 4 years and maximum 12 years. RESULTS: The mean age at Kaposi's sarcoma diagnosis was 39 years and average duration of known HIV seropositivity was 4 years. At Kaposi's sarcoma diagnosis, only 19% patients were on HAART and only 7% patients had an undetectable plasma HIV viral load. Seventy-nine (31%) patients had AIDS clinical Trial Group stage T1 disease at Kaposi's sarcoma diagnosis and 122 (48%) had AIDS clinical Trial Group stage I1 disease (CD4 cell count < 150 cells/microl). Nodular grade Kaposi's sarcoma represented 28% of the tumours and was significantly associated with black African ethnicity and AIDS clinical Trial Group T1 stage disease. The overall 5 year survival is 89% (95% confidence interval 84-93). One hundred and sixty-three patients were treated with HAART alone for T0 stage Kaposi's sarcoma; only one died of Kaposi's sarcoma and only 37 (22%) required chemotherapy, giving a systemic treatment-free survival at 5 years of 74% (95% confidence interval 67 82) and the overall survival at 5 years is 91% (95% confidence interval 87-95). CONCLUSION: The high success rate of HAART in a large cohort of AIDS-Kaposi's sarcoma patients over a prolonged period of follow-up will reassure patients and clinicians that this is a well tolerated and effective approach to stage T0 Kaposi's sarcoma. PMID- 19550284 TI - Metabolic bone disease in HIV infection. PMID- 19550285 TI - HIV integration site distributions in resting and activated CD4+ T cells infected in culture. AB - OBJECTIVE: The goal of this study was to investigate whether the location of HIV integration differs in resting versus activated T cells, a feature that could contribute to the formation of latent viral reservoirs via effects on integration targeting. DESIGN: Primary resting or activated CD4 T cells were infected with purified X4-tropic HIV in the presence and absence of nucleoside triphosphates and genomic locations of integrated provirus determined. METHODS: We sequenced and analyzed a total of 2661 HIV integration sites using linker-mediated PCR and 454 sequencing. Integration site data sets were then compared to each other and to computationally generated random distributions. RESULTS: HIV integration was favored in active transcription units in both cell types, but integration sites from activated cells were found more often in genomic regions that were dense in genes, dense in CpG islands, and enriched in G/C bases. Integration sites from activated cells were also more strongly correlated with histone methylation patterns associated with active genes. CONCLUSION: These data indicate that integration site distributions show modest but significant differences between resting and activated CD4 T cells, and that integration in resting cells occurs more often in regions that may be suboptimal for proviral gene expression. PMID- 19550286 TI - The HIV hide and seek game: an immunogenomic analysis of the HIV epitope repertoire. AB - BACKGROUND: Viruses employ various means to evade immune detection. One common evasion strategy is the removal of CD8 cytotoxic T-lymphocyte (CTL) epitopes. METHOD: Here, we use bioinformatic tools to compute the HIV CTL epitope repertoire presented by over 8000 HIV sequences in multiple Human Leukocyte Antigen alleles. We define the 'Size of Immune Repertoire' (SIR) score, which represents the ratio between the number of the predicted epitopes within a protein and their expected number within a scrambled version of the same protein. RESULTS: We show that HIV proteins present less epitopes than expected and that the number of epitopes gradually decreases from SIV to recent HIV sequences. The decrease of the SIR score of HIV is accompanied by a high frequency of replacement mutations within epitopes. The SIR score of the different HIV proteins is not uniform. The regulatory proteins, Tat and Rev, expressed early during cellular infection have a low SIR score, whereas virion-associated genes that are expressed later, such as Env, Pol and Gag, have a higher SIR score. Actually, the SIR score of Gag keeps increasing over time. CONCLUSION: We hypothesize that our results reflect an HIV immune evasion strategy. This involves the targeting of the CTL immune response to viral structural and enzyme proteins, allowing the virus a time interval to propagate before its host cells are destroyed by CTLs. An efficient anti-HIV CTL response against HIV should thus also target the regulatory genes that HIV seeks to hide from the immune system. PMID- 19550287 TI - Safety, tolerability, and systemic absorption of dapivirine vaginal microbicide gel in healthy, HIV-negative women. AB - OBJECTIVES: To assess the local and systemic safety of dapivirine vaginal gel vs. placebo gel as well as the systemic absorption of dapivirine in healthy, HIV negative women. METHODS: Two prospective, randomized, double-blind, placebo controlled phase I/II studies were conducted at five research centers, four in Africa and one in Belgium. A total of 119 women used dapivirine gel (concentrations of 0.001, 0.002, 0.005, or 0.02%), and 28 used placebo gel twice daily for 42 days. The primary endpoints were colposcopic findings, adverse events, Division of AIDS grade 3 or grade 4 laboratory values, and plasma levels of dapivirine. RESULTS: Safety data were similar for the dapivirine and placebo gels. None of the adverse events with incidence more than 5% occurred with greater frequency in the dapivirine than placebo groups. Similar percentages of placebo and dapivirine gel users had adverse events that were considered by the investigator to be related to study gel. A total of five serious adverse events occurred in the two studies, and none was assessed as related to study gel. Mean plasma concentrations of dapivirine were approximately dose proportional, and, within each dose group, mean concentrations were similar on days 7, 28, and 42. The maximum observed mean concentration was 474 pg/ml in the 0.02% gel group on day 28. Two weeks after the final application of study gel, mean concentrations decreased to 5 pg/ml or less. CONCLUSION: Twice daily administration of dapivirine vaginal gel for 42 days was safe and well tolerated with low systemic absorption in healthy, HIV-negative women suggesting that continued development is warranted. PMID- 19550289 TI - Monitoring HIV vaccine trial participants for primary infection: studies in the SIV/macaque model. AB - OBJECTIVES: The ability to detect and quantify acute HIV-1 infection prior to seroconversion would be an important tool for use in HIV vaccine clinical efficacy trials. We have utilized the SIV/rhesus monkey model to evaluate whether samples more easily obtained than peripheral blood might be used for intensive monitoring of vaccine trial participants. METHODS: We have evaluated viral loads in peripheral blood, saliva, feces, and urine of five rhesus monkeys during primary SIVmac251 infection by quantitative real-time PCR. As an alternative to the direct monitoring of frozen samples, we have also developed a fully quantitative viral load assay utilizing dried blood spots. RESULTS: Although all compartments were found to harbor viral RNA during primary infection, viral RNA could be detected in the peripheral compartments only when levels of plasma viremia exceed a threshold value of 10 RNA copies/ml. We found no direct correlation between viral burden in plasma and saliva, feces, or urine viral loads. Importantly, both dried saliva and whole blood spots can be used for viral detection. Quantitative whole blood or plasma spotting correlated well with viral burden in plasma during both the acute and set point phase of infection. CONCLUSION: Dried blood spots are amenable to rapid quantitative viral load testing. Whole blood spotting has a significant logistical benefit as it requires low blood volumes and no blood processing. Saliva or dried saliva spots or both are potential candidates for acute phase diagnostic screening. These studies indicate the feasibility of intensive monitoring of HIV-1 vaccine trial participants for virus acquisition in resource-limited settings. PMID- 19550290 TI - HIV, the brain, children, HAART and 'neuro-HAART': a complex mix. PMID- 19550288 TI - Longitudinal changes in regional fat content in HIV-infected children and adolescents. AB - BACKGROUND: Alterations in regional fat are often reported in HIV infection. Prior studies have not distinguished between normal changes in regional fat related to sexual maturation and those due to HIV. The study aim was to compare changes in regional fat distribution in HIV-infected (HIV+) and healthy (HIV-) children and adolescents living in the United States. METHODS: Serial dual energy X-ray absorptiometry was performed at baseline and two annual follow-up visits in 64 HIV+ and 147 HIV--participants aged 6-16 years. Total, leg, arm, and trunk fat masses (kg) and regional fat distribution as the percentage of total body fat (%) were compared. RESULTS: HIV+ and HIV--participants did not differ in total fat mass, but the HIV+ group had significantly lower leg and greater arm fat and trunk fat percentage at all time points. Over time, decreases in leg fat percentage and increases in arm fat percentage were more marked among the HIV+ group. Differences between HIV+ and HIV--groups in arm and leg fat percentage remained significant when age, sex, race, height, and pubertal stage were accounted for by mixed effect modeling. Apart from prior treatment with stavudine, no differences in fat distribution were observed according to treatment or degree of immunodeficiency or viremia. CONCLUSION: Although no single pattern of change in regional fat distribution was uniquely associated with HIV, perinatally HIV-infected youth manifest significantly decreased leg fat and increased arm and trunk fat. These differences increase over time and may contribute to cardiovascular disease risk. PMID- 19550291 TI - Hepatitis B and long-term HIV outcomes in coinfected HAART recipients. AB - BACKGROUND: Chronic hepatitis B (CH-B) is common among HIV-infected individuals and increases liver-related mortality in the absence of HAART. The impact of CH-B on long-term HAART outcomes has not been fully characterized. METHODS: To address this question, HAART initiators enrolled in the Multicenter AIDS Cohort Study were retrospectively analyzed. Patients were classified by hepatitis B category based on serology at the time of HAART initiation. The association of CH-B with mortality, AIDS-defining illnesses, CD4 cell rise, and HIV suppression was assessed using regression analysis. RESULTS: Of 816 men followed for a median of 7 years on HAART, 350 were never hepatitis B virus (HBV) infected, 357 had past infection, 45 had CH-B, and 64 were only core-antibody positive. Despite HAART, AIDS-related mortality was the most common cause of death [8.3/1000 person-years (PYs)]. It was highest in those with CH-B (17/1000 PYs, 95% confidence interval 7.3, 42) and lowest among never HBV infected (2.9/1000 PYs, 95% confidence interval 1.4, 6.4). In a multivariable model, patients with CH-B had a 2.7-fold higher incidence of AIDS-related mortality compared with those never infected (P = 0.08). Non-AIDS-related mortality was also highest among those with CH-B (22/1000 PYs), primarily due to liver disease (compared to never infected, adjusted hazard ratio 4.1, P = 0.04). There was no significant difference in AIDS defining events, HIV RNA suppression, and CD4 cell increase. CONCLUSION: In HIV infected patients receiving long-term HAART, HBV status did not influence HIV suppression or CD4 cell increase. However, mortality was highest among those with CH-B and was mostly due to liver disease despite HBV-active HAART. PMID- 19550292 TI - A proteomic study on a human osteosarcoma cell line Saos-2 treated with diallyl trisulfide. AB - Garlic is generally used as a therapeutic reagent against various diseases, and numerous studies have indicated that garlic and its derivatives can reduce the risk of various types of human cancer. Diallyl trisulfide (DATS), a major member of garlic derivatives, could inhibit the cell proliferation by triggering either cell cycle arrest or apoptosis in a variety of cancer cell lines as shown in many studies. However, whether DATS has the same effect on human osteosarcoma cells remains unknown. In this study, we have attempted to analyze the effects of DATS on cell proliferation, cell cycle, induction of apoptosis, global protein expression pattern in a human osteosarcoma cell line Saos-2 cells, and the potential molecular mechanisms of the action of DATS. Saos-2 cells, a human osteosarcoma cell line, were treated with or without 25, 50, and 100 micromol/l DATS for various time intervals. The cell proliferation, cell cycle progression, and apoptosis were examined in this study. Then, after treatment with or without 50 micromol/l DATS for 48 h, protein add pattern in Saos-2 cells were systematically studied using two-dimensional electrophoresis and mass spectrometry. DATS could inhibit the proliferation of Saos-2 cells in a dose dependent and time-dependent manner. Moreover, the percentage of apoptotic cell and cell arrest in G0/G1 phase was also dose-dependent and time-dependent upon DATS treatment. A total of 27 unique proteins in Saos-2 cells, including 18 downregulated proteins and nine upregulated proteins, were detected with significant changes in their expression levels corresponding to DATS administration. Interestingly, almost half of these proteins (13 of 27) are related to either the cell cycle or apoptosis. DATS has the ability to suppress cell proliferation of Saos-2 cells by blocking cell cycle progression and inducing apoptosis in a dose and time-dependent manner. The proteomic results presented, therefore, provide additional support to the hypothesis that DATS is a strong inducer of apoptosis in tumor cells. However, the exact molecular mechanisms, how these proteins significantly changed in the Saos-2 cell line upon DATS treatment, should be further studied. PMID- 19550293 TI - Smac mimetics as new cancer therapeutics. AB - The recent discovery of Smac and the elucidation of its structure and function have led to the rapid development of Smac mimetics, comprising Smac derivative and mimicking molecules, for use in cancer treatment. Smac is an endogenous proapoptotic protein that resides in the mitochondria and is released when a cell is triggered to undergo programmed cell death. One of the mechanisms by which Smac promotes apoptosis is through its ability to inhibit inhibitors of apoptosis (IAPs), by direct inhibition and/or proteasomal degradation of some members of the IAP family, and therefore disinhibit caspases. Thus, the use of Smac mimetics as anticancer agents follows a rational approach in cancer therapeutics. This approach directly targets dysregulated, neoplastic cells that overexpress IAPs or underexpress Smac. Although Smac mimetics are able to elicit an anticancer response when used alone, these molecules can also function effectively and synergistically when combined with other therapeutic agents. A variety of Smac mimetic types comprising peptides, polynucleotides, and compounds have been studied both in vitro and in vivo. This discussion addresses the current status of Smac mimetics in cancer research. PMID- 19550294 TI - Folate-bearing doxorubicin-loaded magnetic poly(N-isopropylacrylamide) microspheres as a new strategy for cancer therapy. AB - Doxorubicin is a classic anticancer agent. Recently, numerous strategies have been used to enhance efficacy of drug delivery for cancer treatment. For example, by modifying poly(N-isopropylacrylamide) microspheres, a nanocarrier, makes it more effective. Conjugation with folic acid increases specific targeted drug delivery towards folate receptor-bearing cancer cells to improve anticancer effectiveness by increasing the tissue's local concentration of drugs. In the current studies, we synthesized folate-bearing, doxorubicin-loaded, magnetic, poly(N-isopropylacrylamide) microspheres (FDMPM) to treat breast cancer cells (human SKBR-3). We found efficiency of drug encapsulation very high (95%) at pH above 7.4. Reverse transcription-PCR showed that cancer cells highly expressed folate receptors. Confocal laser scanning microscopy and flow cytometry revealed internalization of the carrier by SKBR-3 in treatments with FDMPM, which was not the case with any other combination for drug delivery (MPM, FMPM, and DMPM). Similarly, SKBR-3 cell growth was inhibited more (assessed by methylthiazolyldiphenyl-tetrazolium bromide and trypan blue exclusion assays) when treated with FDMPM than with any other combinations. Current results confirm our predication and demonstrate that FDMPM has potential as a new targeting strategy in cancer therapy. PMID- 19550295 TI - Effect of cytotoxic agents and retinoic acid on Myc-N protein expression in neuroblastoma. AB - AIM: Neuroblastoma is an important pediatric tumor in which Myc-N amplification is a well-known poor prognostic indicator. It has a great diversity in clinical behavior. The effect of pharmacologic agents used in neuroblastoma treatment on Myc-N expression is still unclear. METHOD: We analyzed Myc-N expression changes by immunocytochemistry in Myc-N-positive Kelly human neuroblastoma cell line using retinoic acid and cytotoxic drugs (cisplatin, vincristine, cyclophosphamide, etoposide, and doxorubicin) and their combinations compared with control conditions. First, concentration of drugs were determined as LD50 doses. Kelly cells and drugs were incubated for 24 hours in 5% CO2, 37 degrees C in 96-well plates. Myc-N expression was scored semiquantitatively as negative, mild, moderate, or high positive. RESULTS: Myc-N amplification did not change with any agent or combination. It was higher than 20 copies in all conditions. Myc-N protein expression was high in control and doxorubicin group. It was moderate in retinoic acid, cyclophosphamide, retinoic acid combined with cyclophosphamide and retinoic acid combined with doxorubicin groups. The expression was mild in cisplatin, vincristine, etoposide, retinoic acid combined with etoposide, and retinoic acid combined with cisplatin groups. Myc-N expression was negative in retinoic acid combined with vincristine group. CONCLUSIONS: Myc-N expression is reduced with cytotoxic agents and retinoic acid in neuroblastoma although Myc-N amplification remains the same. Retinoic acid combined with vincristine is the most effective combination to reduce Myc-N expression. Our results suggest that therapeutic applications of these agents as low dose maintenance therapy might be useful. PMID- 19550296 TI - Determining tissue of origin for metastatic cancers: meta-analysis and literature review of immunohistochemistry performance. AB - BACKGROUND: Pathologists use various panels of immunohistochemical (IHC) stains to identify the site of tissue of origin for metastatic tumors, particularly poorly or undifferentiated cancers of unknown or uncertain origin. Although clinicians believe that immunostains contribute greatly to determining the probable primary site among 3 or more possibilities, objective evidence has not been convincingly presented. This meta-analysis reviews the objective evidence supporting this practice and summarizes the performance reported in 5 studies published between 1993 and 2007. METHODS: A literature search was conducted to identify IHC performance studies published since 1990 that were masked, included more than 3 tissues types, and used more than 50 specimens. The 5 studies found in this search were separated into 2 subgroups for analysis: those, which included only metastatic tumors (n = 368 specimens) and the blended studies, which combined primary tumors and metastases (n = 289 specimens). RESULTS: The meta-analysis found that IHCs provided the correct tissue identification for 82.3% (95% confidence interval = 77.4%-86.3%) of the blended primary and metastatic samples and 65.6% (95% confidence interval = 60.1%-70.7%) of metastatic cancers. This difference is both clinically and statistically significant. CONCLUSIONS: This literature review confirms that there is still an unmet medical need in identification of the primary site of metastatic tumors. It establishes minimum performance requirements for any new diagnostic test intended to aid the pathologist and oncologist in tissue of origin determination. PMID- 19550297 TI - Utility of fascin and JunB in distinguishing nodular lymphocyte predominant from classical lymphocyte-rich Hodgkin lymphoma. AB - Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) and lymphocyte-rich classical Hodgkin lymphoma (LRCHL), although clinically and morphologically similar, differ biologically and in prognosis. Immunolabeling of Reed-Sternberg (RS) cells in LRCHL and lymphocytic and/or histiocytic variants (L&H cells) in NLPHL is often required to help distinguish between the 2 variants. Our aim was to evaluate fascin (a distinct 55-kd actin-bundling protein) and JunB (an activator protein-1 family transcription factor) to differentiate NLPHL from LRCHL. A total of 35 archival cases of NLPHL (n = 24) and LRCHL (n = 11) from adults and children were studied. Slides were reviewed for all cases and clinical, morphologic, and immunohistochemical features were evaluated. Each case was immunostained for fascin and JunB, and immunoreactivity of RS cells, L&H cells, and background lymphocytes were recorded. Whereas occasional L&H cells were weakly positive for fascin in 3 out of 24 (12.5%) cases of NLPHL, RS cells in LRCHL were positive for fascin in 11 out of 11 (100%) cases with a strong cytoplasmic staining pattern. JunB was positive in 10 out of 24 (41.7%) of NLPHL cases, and 11 out of 11 (100%) of LRCHL cases, showing a stippled and/or diffuse nuclear staining pattern. In addition to L & H Cells, JunB also stained small background lymphocytes, particularly in areas of progressively transformed germinal centers of NLPHL. Either stains when tested alone, if negative, or with rare L&H cell weak positivity for fascin, is indicative of NLPHL. The L&H cells of NLPHL cases were negative for concomitant staining in 24 out of 24 (100%) cases. Concomitant positive staining of classic RS cells for fascin and JunB was found in 11 out of 11 (100%) of LRCHL cases. Although fascin positivity alone supports the diagnosis of LRCHL, concomitant positivity offers stronger support and is less likely to lead to a false conclusion if aberrant fascin staining were to be encountered in a case of NLPHL. Staining for fascin and JunB provides a basis for distinguishing NLPHL from LRCHL and offers an alternative to other antibody profiles. PMID- 19550298 TI - Influence of home blood pressure measuring conditions in the evening on the morning-evening home blood pressure difference in treated hypertensive patients: the J-HOME study. AB - OBJECTIVE: The objective of this study was to clarify the factors affecting the morning-evening home systolic blood pressure (BP) difference (home systolic ME dif) in treated hypertensive patients, including evening home BP measuring conditions, based on the data from the Japan Home versus Office BP Measurement Evaluation study. METHODS: The study participants were 3303 essential hypertensive patients (mean age 66.2+/-10.5 years; females 55.3%) treated using antihypertensive drug therapy in primary care settings in Japan. Multivariate regression analysis including the variables that were significantly associated with the home systolic ME dif on the univariate analysis was performed. RESULTS: The mean of the home systolic/diastolic ME dif was 6.1+/-10.8/4.8+/-6.5 mmHg. The proportion of patients measuring evening BP after drinking alcohol was 20.5%, and the proportion of patients measuring evening BP after bathing was 76.8%. Uncontrolled morning systolic BP (morning systolic BP > or =135 mmHg), controlled evening systolic BP (evening systolic BP <135 mmHg), older age (> or =65 years), measurement of evening BP after drinking, and measurement of evening BP after bathing were positively associated with the home systolic ME dif on multivariate regression analysis. CONCLUSION: Measurement of evening BP after drinking and measurement of evening BP after bathing were strongly associated with an increased home systolic ME dif, independent of morning and evening home BP levels. Therefore, evening home BP measuring conditions (before or after drinking alcohol and bathing) should be taken into account while evaluating the home systolic ME dif. PMID- 19550299 TI - Renal cystic dysplasia, paucity of bile ducts, situs inversus, bowing of the femora in two siblings in the Reunion Island: a ciliopathy? PMID- 19550301 TI - Mast cells: makers and breakers of allergic inflammation. AB - PURPOSE OF REVIEW: Recent findings have developed, changed, challenged and extended the established view of mast cell biology and of their role in inflammation. The purpose of this review is to present and discuss recent research in this area relating to allergic inflammation. RECENT FINDINGS: Mast cells are traditionally viewed only as IgE-activated histamine-providing effector cells of allergic inflammation. Yet, recent findings also support a role for mast cells in the sensitization to allergens, and early released human mast cell tryptase has been shown to break down IgE, hence breaking the key maker of allergic inflammation. Furthermore, mast cells demonstrate proinflammatory action through nonhistamine and non-IgE-mediated routes. For example, the role of platelet-activating factor in contributing to allergic inflammation and anaphylaxis in mice is now starting to be shown in humans. Most surprisingly, mast cells have also emerged as modulators and downregulators of allergic inflammation. For example, the newly discovered siglec-8 receptor might act as an inhibitor of mast cell degranulation and thus of mast cell-mediated allergic inflammation. Also, mast cell-derived IL-10 is shown to be a modulator of allergic inflammation, demonstrated in a murine model of contact hypersensitivity. SUMMARY: Understanding the mast cell's divergent phenotypes based on situation and setting is required to truly discover their role in allergic inflammation. PMID- 19550303 TI - Brachial plexus block with or without ultrasound guidance. AB - PURPOSE OF REVIEW: Should ultrasound or nerve stimulation be used for brachial plexus blocks? We investigated last year's literature to help answer this question. RECENT FINDINGS: Many of the reports concluded that ultrasound guidance may provide a higher success rate for brachial plexus blocks than guidance by nerve stimulator. However, the studies were not large enough to conclude that ultrasound will reduce the risk of nerve injury, local anesthetic toxicity or pneumothorax. Ultrasound may reveal anatomical variations of importance for performing brachial plexus blocks. For postoperative analgesia, 5 ml of ropivacaine 0.5% has been sufficient for an ultrasound-guided interscalene block. For peroperative anesthesia, as much as 42 ml of a local anesthetic mixture was calculated to be appropriate for an ultrasound-guided supraclavicular method. For the future, we notice that three-dimensional and four-dimensional ultrasound technology may facilitate visualizing the needle, the nerves and the local anesthetic distribution. Impedance measurements may be helpful for nerve blocks not guided by ultrasound. SUMMARY: We think that the literature gives a sufficient basis to recommend the use of ultrasound for guidance of brachial plexus blocks. The potential for ultrasound to improve efficacy and reduce complications of brachial plexus blocks requires larger scaled studies. PMID- 19550304 TI - Regional anesthesia and obesity. AB - PURPOSE OF REVIEW: Worldwide, the number of overweight and obese patients has increased dramatically. As a result, anesthesiologists routinely encounter obese patients daily in their clinical practice. The use of regional anesthesia is becoming increasingly popular for these patients. When appropriate, a regional anesthetic offers advantages and should be considered in the anesthetic management plan of obese patients. The following is a review of regional anesthesia in obesity, with special consideration of the unique challenges presented to the anesthesiologist by the obese patient. RECENT FINDINGS: Recent studies report difficulty in achieving peripheral and neuraxial blockade in obese patients. For example, there is an increased incidence of failed blocks in obese patients compared with similar, normal weight patients. Despite difficulties, regional anesthesia can be used successfully in obese patients, even in the ambulatory surgery setting. SUMMARY: Successful peripheral and neuraxial blockade in obese patients requires an anesthesiologist experienced in regional techniques, and one with the knowledge of the physiologic and pharmacologic differences that are unique to the obese patient. PMID- 19550305 TI - Regional anesthesia and the patient with preexisting neurological disease. PMID- 19550302 TI - Abnormal skin barrier in the etiopathogenesis of atopic dermatitis. AB - PURPOSE OF REVIEW: Many recent studies have revealed the key roles played by Th1/Th2 cell dysregulation, IgE production, mast cell hyperactivity, and dendritic cell signaling in the pathogenesis of atopic dermatitis. Accordingly, current therapy has been largely directed towards ameliorating Th2-mediated inflammation and/or pruritus. We will review here emerging evidence that the inflammation in atopic dermatitis results from inherited and acquired insults to the barrier and the therapeutic implications of this new paradigm. RECENT FINDINGS: Recent molecular genetic studies have shown a strong association between mutations in FILAGGRIN and atopic dermatitis, particularly in Northern Europeans. But additional acquired stressors to the barrier are required to initiate inflammation. Sustained hapten access through a defective barrier stimulates a Th1 --> Th2 shift in immunophenotype, which in turn further aggravates the barrier. Secondary Staphylococcus aureus colonization not only amplifies inflammation but also further stresses the barrier in atopic dermatitis. SUMMARY: These results suggest a new 'outside-to-inside, back to outside' paradigm for the pathogenesis of atopic dermatitis. This new concept is providing impetus for the development of new categories of 'barrier repair' therapy. PMID- 19550306 TI - The Mediterranean diet revisited: evidence of its effectiveness grows. AB - PURPOSE OF REVIEW: The optimal dietary strategy for the prevention of chronic degenerative diseases remains a challenging and a highly relevant preventive health issue. In recent years, there have been ongoing investigations of the effectiveness of the Mediterranean diet in preventing the development of cardiovascular and other chronic diseases. Initial research suggested that the consumption of diets high in vegetable oil, fish, and complex grains and carbohydrates conferred a benefit in reducing the prevalence of cardiovascular disease. The purpose of this review is to examine the evidence in support of the effectiveness of the Mediterranean diet in reducing the prevalence of cardiovascular and chronic disease and their associated risk factors. RECENT FINDINGS: Recent studies intended to evaluate the Mediterranean diet in the primary or secondary prevention of cardiovascular and other chronic degenerative diseases have been focused on examining the impact of a whole dietary approach rather than on isolating single nutrients; it is recognized that analyses of single nutrients ignore the important and complex interactions between components of a diet and, more importantly, because people do not eat isolated nutrients. Therefore, dietary scores estimating the adherence to a Mediterranean diet within a population have been developed. Consumption of a Mediterranean diet has been found to be associated with a reduction of overall mortality and a reduced incidence of cardiovascular disease. SUMMARY: The Mediterranean diet has been consistently shown to be associated with favourable health outcomes and a better quality of life. The findings suggest that significant health gains can be expected in a general population whose diet reflects the nutritional principles reflected in a Mediterranean diet. PMID- 19550307 TI - A tragic triad: coronary artery disease, nicotine addiction, and depression. AB - PURPOSE OF REVIEW: Despite the availability of multiple resources for treating smoking in patients with cardiovascular disease (CVD) and the well known risks of continued smoking, a majority of smokers resume smoking after hospitalization for acute CVD. Depression is common among patients with CVD and is associated with failure to follow recommendations to reduce cardiac risk. This review examines the complex relationship between smoking and depression in patients with CVD and current evidence for treating this triad. RECENT FINDINGS: The prevalence of depressive symptoms among smokers hospitalized with CVD is 22-24%. Smokers with depressive symptoms are more likely to return to smoking after hospital discharge compared with nondepressed smokers. Stronger nicotine withdrawal symptoms among the depressed smokers contribute to relapse. Secondary analyses suggest that bupropion SR and cognitive behavioral therapy may be effective treatments for smokers with depression and CVD. A systematic review of smoking interventions in hospitalized patients found that only intensive counseling interventions with follow-up for more than 1 month were effective for smokers with CVD. SUMMARY: Clinicians should consider screening all smokers with CVD for depression, particularly during hospitalization for an acute event. Smokers hospitalized with CVD need intensive counseling lasting more than 1 month after discharge, and smokers with depressive symptoms need even more prolonged treatment for quitting. The addition of pharmacotherapy to long-term counseling has the potential to further improve cessation rates. PMID- 19550308 TI - Medication adherence and the patient with coronary artery disease: challenges for the practitioner. AB - PURPOSE OF REVIEW: Adherence to secondary prevention medications among patients with coronary artery disease is essential in minimizing risks of recurrent myocardial infarction and mortality. Despite its importance, nonadherence remains a significant problem among this population, and a variety of studies have sought to determine its prevalence, contributing factors, and interventions for improvement. RECENT FINDINGS: Several recent studies have demonstrated improving rates of adherence over time, though the overall prevalence of nonadherence remains significant. Other studies have identified important factors associated with nonadherence, and two recent trials tested interventions to improve adherence rates. SUMMARY: Although there have been some improvements in adherence rates, it remains a significant issue. Nonadherence increases both general and cardiac-specific adverse events. Several important factors such as patient attitudes, external influences, concurrent comorbidities, and health system characteristics appear to significantly impact adherence rates. Recent trials to improve adherence rates have demonstrated only modest effects, but lessons from these initial interventions should be incorporated into future strategies to improve adherence. PMID- 19550309 TI - Preventing future acute coronary events: is the target the so-called vulnerable plaque or the high-risk or vulnerable patient? AB - PURPOSE OF REVIEW: Heart disease still remains the leading cause of mortality in the USA in spite of recent reductions in the death rate; complications of coronary artery disease and its sequelae are the most common mechanism of demise. Although there have been great advances in the prevention and treatment of acute myocardial infarction, greater emphasis on prevention will likely be needed to reduce acute coronary complications further. RECENT FINDINGS: The literature is replete with articles on the attempted localization of so-called vulnerable plaques and vulnerable, or high-risk patients. They emphasize the importance of the identification of that high-risk plaque or high-risk individual prior to a subsequent coronary event. This article highlights the breakthroughs into the pathophysiology of acute coronary syndromes in the past few decades and presents a perspective on current treatments, improved risk stratification and potential technological advances that may impact diagnosis and management. SUMMARY: Unfortunately, the search for the so-called vulnerable plaque is hampered by the lack of both natural history studies and proven local or regional therapies for these otherwise asymptomatic plaques. Thus, emphasis on the vulnerable or high risk patient is appropriate, but identifying these individuals in primary prevention is also fraught with difficulty. No specific recommendations can be made at present, as more data are needed in both areas. However, guidelines for future advances are proposed. PMID- 19550310 TI - Three-dimensional echocardiography for assessment of mitral valve stenosis. AB - PURPOSE OF REVIEW: Since the last few years, three-dimensional echocardiography (3DE) has become an accurate tool for mitral stenosis assessment. We will review the latest developments of 3DE in this matter. RECENT FINDINGS: Accuracy of 3DE planimetry is superior to the accuracy of the invasive Gorlin's method for mitral valve area (MVA) measurements when a median value obtained from two-dimensional planimetry, pressure half-time, and proximal isovelocity surface area method is used as the gold standard. 3DE improves MVA measurement particularly in less experienced operators compared with experienced operators. 3DE also improves the measurement of MVA in patients with calcific mitral stenosis by means of colour planimetry of the flow stream. Comparison of mitral valve volumes measured by 3DE in patients with critical and without critical stenosis has shown significantly larger volumes in patients with critical stenosis. SUMMARY: Currently, there is sufficient evidence that 3DE is superior to two-dimensional echocardiography and may be routinely used in the quantification of the MVA in mitral stenosis. In the coming years, 3DE might replace Gorlin's method as the gold standard for MVA quantification and may eventually make cardiac catheterization unnecessary. PMID- 19550311 TI - 3D echocardiography: evaluation of the right ventricle. AB - PURPOSE OF REVIEW: Evaluation of right ventricular (RV) volume, function and mass has been challenging because of its unique cavity geometry. However, it is indispensable, especially in patients with signs of right-sided heart failure and those with congenital heart diseases. Thus, many investigations on the RV have been published to this end. Although clinical investigations of the RV have been focused on the use of 2-dimenisonal (2D) echocardiography, progressive development of three-dimensional (3D) echocardiography has made it possible to evaluate the RV in clinical settings lately. RECENT FINDINGS: Accuracy of newly developed real-time 3D echocardiography for determining RV volume and function has been reported with the use of magnetic resonance imaging (MRI). A slight but significant underestimation of MRI derived RV volumes by 3D echocardiographic methods have been found in multiple recent human studies. One of the in-vitro studies rightly reported the importance of technical factors, causing the inaccuracy of 3D echocardiography. SUMMARY: Despite the development of new real time 3D echocardiographic systems, there still exist limitations in clinical settings due to technical factors. Thus, the combination of conventional 2D and Doppler methods and 3D echocardiography is recommended in the evaluation of the RV at the present time. PMID- 19550312 TI - Mechanisms of insulin resistance assessed by dynamic in-vivo positron emission tomography imaging. AB - PURPOSE OF REVIEW: Skeletal muscle insulin resistance is a hallmark characteristic of type 2 diabetes, although the exact causes of insulin resistance are unknown. In-vivo methods to assess mechanisms that determine insulin resistance in humans are critical to improve our understanding of insulin resistance in obesity and type 2 diabetes. In this review, we examine recent studies utilizing dynamic in-vivo PET imaging in assessing insulin resistance in humans. RECENT FINDINGS: PET imaging of glucose metabolism in vivo has revealed novel and important information about the regulation of glucose metabolism in skeletal muscle. Using dynamic PET imaging, studies have impairments in glucose metabolism at multiple sites, including delivery, phosphorylation, and transport within skeletal muscle. Impairments in glucose phosphorylation as well as glucose transport defects may play an important role in understanding the disorder of skeletal muscle insulin resistance. SUMMARY: PET imaging has great potential to yield significant and promising insight into insulin resistance in skeletal muscle. Dynamic in-vivo PET imaging can provide valuable information regarding the mechanisms and specific loci of skeletal muscle insulin resistance in humans. PMID- 19550313 TI - Methods for assessing intrahepatic fat content and steatosis. AB - PURPOSE OF REVIEW: Intrahepatic fat content is increasingly being recognized as an integral part of metabolic dysfunction. This article reviews available methods for the assessment of hepatic steatosis. RECENT FINDINGS: Apart from liver biopsy, there are several noninvasive radiologic modalities for evaluating nonalcoholic fatty liver disease. Ultrasonography, computed tomography, and traditional MRI remain largely qualitative methods for detecting mild to severe degrees of steatosis rather than quantitative methods for measuring liver fat content, even though novel attempts to collect objective quantitative information have recently been developed. Still, their sensitivity at mild degrees of steatosis is poor. Undoubtedly, most methodological advances have occurred in the field of MRI and magnetic resonance spectroscopy, which currently enable the accurate quantification of intrahepatic fat even at normal or near normal levels. Xenon computed tomography was also recently shown to offer another objective tool for the quantitative assessment of steatosis, although more validation studies are required. SUMMARY: Several modalities can be used for measuring intrahepatic fat and assessing steatosis; the choice will ultimately depend on the intended use and available resources. PMID- 19550314 TI - Photodynamic therapy for cholangiocarcinoma: overview and new developments. AB - PURPOSE OF REVIEW: Photodynamic therapy (PDT) with hematoporphyrins has emerged as promising treatment for nonresectable cholangiocarcinoma in several prospective observational studies and two randomized studies. This review describes the mechanism of action of PDT, gives an overview of clinical experience in cholangiocarcinoma and summarizes the results published in 2007 and 2008. RECENT FINDINGS: The mechanism of action of PDT has been further elucidated. PDT induces an apoptotic, antiangiogenic as well as an immunomodulatory response. Interleukin-6, a bile duct epithelium growth factor correlating with tumor burden, decreases after PDT. The efficacy of PDT was confirmed in a comparative study in the United States. Patients with no visible mass on imaging studies, high serum albumin levels and treatment immediately after diagnosis seem to benefit most from PDT. Although it is recommended to perform PDT in bile ducts without stents in place, illumination through metal stents is possible if the light dose is adjusted. Meso-tetrahydroxyphenyl chlorine is a new potent photosensitizer for PDT of cholangiocarcinoma. SUMMARY: In advanced nonresectable cholangiocarcinoma, PDT is the only evidence-based treatment that improves survival when compared with stenting. Therefore, PDT should be offered to those who are unsuitable for surgery. PMID- 19550315 TI - Pancreatic surgery. AB - PURPOSE OF REVIEW: Surgery of the pancreas is evolving as the understanding of pancreatic disease improves. This report reviews the work published over the last year related to pancreatic surgery and the diseases addressed by surgical techniques. RECENT FINDINGS: Obesity is an important risk factor for many diseases of the pancreas, including pancreatic adenocarcinoma. Recent evidence suggests that obese patients with pancreatic cancer appear to have more advanced disease at the time of diagnosis and a worse outcome following resection. The issues surrounding adjuvant treatment for pancreatic cancer with chemotherapy and/or radiation therapy continue to be evaluated. Cystic lesions of the pancreas remain a vexing treatment dilemma, but as we learn more about their natural history, more thoughtful recommendations for management become possible. Resection of pancreatic endocrine neoplasms is often appropriate, even in the face of metastatic disease. Minimally invasive approaches to the drainage of infected pancreatic necrosis are beginning to gain acceptance. The pain of chronic pancreatitis may be lessened by operative intervention and possibly radiation. SUMMARY: Each year more is learned about the natural history of pancreatic lesion. For those dedicated to the study and treatment of this gland, several new advances help the clinician with treatment decisions. PMID- 19550316 TI - Anticoagulants to prevent placenta-mediated pregnancy complications: a review of current evidence. AB - PURPOSE OF REVIEW: Placenta-mediated pregnancy complications are relatively common and are associated with significant morbidity and mortality. Although anticoagulants are increasingly being used to prevent these complications, the evidence supporting their use is limited. This article will focus on reviewing the current evidence base for the use of anticoagulants to prevent placenta mediated pregnancy complications in women with or without identifiable thrombophilia. RECENT FINDINGS: Controversy exists whether inherited or acquired thrombophilias cause placenta-mediated complications. Small randomized studies with methodological limitations suggest that antepartum anticoagulant prophylaxis will reduce pregnancy loss in women with both acquired and inherited thrombophilia and prior pregnancy loss. There are no published randomized controlled trials examining anticoagulant prophylaxis in thrombophilic women with prior preeclampsia, intrauterine growth restriction or placental abruption. More recently, the benefit of antepartum anticoagulant prophylaxis in women without thrombophilia with prior placenta-mediated pregnancy complications has been suggested in a small pilot randomized trial. SUMMARY: Overall, although antepartum anticoagulant prophylaxis has the potential to reduce placenta mediated pregnancy complications in women with and without identifiable thrombophilia, the data generated by these trials are methodologically limited and inadequate. Hence, we conclude that further trials are required prior to adopting the use of antepartum anticoagulant prophylaxis to prevent placenta mediated pregnancy complications in routine clinical practice. PMID- 19550317 TI - New P2Y12 antagonists. AB - PURPOSE OF REVIEW: The P2Y12 antagonist clopidogrel has a well established role as an antithrombotic agent in the settings of percutaneous coronary intervention and acute coronary syndromes. However, clopidogrel has a number of disadvantages that have led to the development of new P2Y12 antagonists. This article reviews the current status of clopidogrel and these new P2Y12 antagonists. RECENT FINDINGS: A poor response to clopidogrel as measured by in-vitro platelet function assays is associated with a poor clinical response to clopidogrel. A patient's response to clopidogrel is partly dependent on factors that decrease (reduced-function CYP2C19 allele; omeprazole) or increase (cigarette smoking) the metabolism of clopidogrel through cytochrome P450 in the liver. Novel P2Y12 antagonists (prasugrel, ticagrelor, cangrelor, and elinogrel) that have advantages over clopidogrel - including more rapid, less variable, and more complete inhibition of platelet function - are in various phases of development. SUMMARY: The clinical benefit of changing clopidogrel treatment based on platelet function tests, single-nucleotide polymorphisms, or both is as yet unproven. Novel P2Y12 antagonists are under investigation to determine whether they can result in better, more rapid, or both, antithrombotic effects than clopidogrel, without an unacceptable increase in hemorrhagic or other side effects. PMID- 19550318 TI - Oral anticoagulation with factor Xa and thrombin inhibitors: on the threshold of change. AB - PURPOSE OF REVIEW: Since the discovery of vitamin K antagonists in the early 1940s, there has been little change in anticoagulation until, in the 1990s, widespread use of low-molecular-weight heparin. Within the next few years, the treatment of arterial and venous thromboembolism is again poised to undergo a major change with the introduction of new oral anticoagulants that are likely to fulfill many of the unmet needs of current warfarin therapy. New drug development has focused on inhibiting specific coagulation factors, with those targeting thrombin and factor Xa being most advanced in development. RECENT FINDINGS: Several landmark studies are now available on the direct thrombin inhibitor, dabigatran etexilate, and the two factor Xa inhibitors, rivaroxaban and apixaban. Recently, dabigatran etexilate received European approval for venous thromboembolism prevention following orthopedic surgery. Rivaroxaban is currently also approved in Europe and Canada for venous thromboembolism prevention in orthopedic surgery, with US Food and Drug Administration approval expected in 2009. SUMMARY: New oral anticoagulant approval may provide safer and easier venous thromboembolism prevention and treatment than warfarin. As we stand on this threshold, this article reflects on anticoagulation breakthroughs, summarizes recent studies, and discusses potential drawbacks. PMID- 19550319 TI - Animal models of thrombosis. AB - PURPOSE OF REVIEW: The thrombotic response to vascular injury is an important clinical problem that mediates most vascular disease complications. Thrombus formation involves an integrated response that is influenced by blood flow, multiple cell types, and numerous circulating factors. As a result, modeling of this complex response using in-vitro or in-silico strategies is insufficient. The use of animal models of thrombosis provides a critical tool for the discovery and initial testing of novel therapies for vascular thrombosis. RECENT FINDINGS: The literature from 2008 to the present provides significant advances in regard to novel models of arterial thrombosis, novel mechanisms underlying thrombus formation, new models and mechanisms related to thrombotic stroke, and preclinical advances in therapeutics for vascular thrombosis. SUMMARY: The formation of occlusive thrombi is complex, involving the integration of many molecular interactions and cell types at the site of vascular injury. The identification of strategies to suppress occlusive thrombus formation without undermining normal hemostatic function is the primary goal of this area of study. PMID- 19550320 TI - The platelet proteome. AB - PURPOSE OF REVIEW: The proteome is the pool of proteins expressed at a given time and circumstance. The word 'proteomics' summarizes several technologies for visualization, quantitation and identification of these proteins. Recent advances in these techniques are helping to elucidate platelet processes which are relevant to bleeding and clotting disorders, transfusion medicine and regulation of angiogenesis. RECENT FINDINGS: Over 1100 platelet proteins have been identified using proteomic techniques. Various subproteomes have been characterized, including platelet releasates (the 'secretome'), alpha and dense granules, membrane and cytoskeletal proteins, platelet-derived microparticles, and the platelet 'phosphoproteome'. Proteomic data about platelets have become increasingly available in integrated databases. SUMMARY: Proteomic experiments in resting and activated platelets have identified novel signaling pathways and secreted proteins which may represent therapeutic targets, as well as potential cancer biomarkers. PMID- 19550321 TI - A logical approach to surgical thromboprophylaxis. AB - PURPOSE OF REVIEW: Venous thromboembolism is an important complication for patients undergoing surgery. Thromboprophylaxis reduces thromboembolic events. RECENT FINDINGS: In this review we discuss the evidence supporting the use of newer agents for thromboprophylaxis, rivaroxaban and dabigatran etexilate. We will also discuss current thromboprophylaxis options for patients undergoing bariatric surgery, including dose and duration of pharmacological prophylaxis and mechanical methods. The most recent The American College of Chest Physicians and American Academy of Orthopedic Surgeons guidelines are also reviewed. SUMMARY: Novel agents may modify our approach to thromboprophylaxis; although these agents offer reduced rates of venographic deep vein thrombosis, their impact on patient important outcomes (clinical deep vein thrombosis, pulmonary embolism and death) requires additional study. PMID- 19550322 TI - Vena caval filters: current knowledge, uncertainties and practical approaches. AB - PURPOSE OF REVIEW: Inferior vena caval (IVC) interruption has been used as a method to prevent pulmonary embolism since the 1940s. Despite an exponential increase in IVC filter use in both the treatment and prophylaxis of venous thromboembolism, there is little evidence to support current practice. This review will discuss controversies related to IVC filters and will provide a practical approach to their use. RECENT FINDINGS: Current practice guidelines recommend that IVC filters be placed in patients with acute proximal deep vein thrombosis and a contraindication to anticoagulation. We do not recommend IVC filters as primary thromboprophylaxis, even for high-risk surgical or trauma patients. We also do not believe that there is a role for IVC filters in cancer patients with venous thromboembolism when traditional anticoagulation has failed. IVC filters have been shown to be associated with an increased risk of recurrent deep vein thrombosis. SUMMARY: IVC filters are indicated in only a small proportion of patients who have venous thromboembolism. In these situations, retrievable filters are recommended. Anticoagulation should be initiated after filter placement as soon as it is safe to do so and the filter should then be removed shortly thereafter. PMID- 19550323 TI - Variation in paraoxonase-1 activity and atherosclerosis. AB - PURPOSE OF REVIEW: Paraoxonase-1 (PON1) is an HDL-associated protein of 354 amino acids with a molecular mass of 43 000 Da. It is synthesized in the liver, and in serum it is almost exclusively associated with HDL. PON1 has been reported to be an important contributor to the antioxidant and anti-inflammatory activities of HDL. PON1 impedes oxidative modification of LDL. PON1 serum activity is related to systemic lipid peroxidation stress and prospective cardiovascular risk. In this review, we discuss the relationship between PON1 activity and atherosclerotic diseases and various factors modulating PON1 activity including genes, age, lifestyle factors and medical conditions. Finally, evidence that pharmacological agents may affect PON1 activity is summarized. RECENT FINDINGS: There is increasing evidence from both animal and human studies linking low PON1 activity to an increased likelihood of cardiovascular diseases. Two prospective studies reported a significantly lower incidence of major cardiovascular events in participants with the highest systemic PON1 activity, compared with those with the lowest activity. SUMMARY: PON1 is a potentially antiatherogenic HDL associated enzyme that protects LDL from oxidative modification. Enhancing PON1 activity could be an important target for future pharmacological agents aimed at decreasing cardiovascular risk. PMID- 19550324 TI - New insights and therapies for the metabolic consequences of Alstrom syndrome. AB - PURPOSE OF REVIEW: Recent studies have begun to evaluate the heterogeneity of insulin resistance in syndromes associated with type 2 diabetes, dyslipidaemia and associated cardiac, renal and hepatic consequences. These insights are of particular importance in Alstrom syndrome in which all of these conditions coexist from a young age with considerable morbidity and reduction in life expectancy. Clear definition of the phenotype in the syndrome may clarify biochemical pathways of crucial importance in propensity to diabetic complications and heart disease in the general population. This review will focus on ways in which more effective treatments can be put in place for Alstrom families from childhood and adolescence. RECENT FINDINGS: Studies of body fat distribution, insulin resistance, glucose metabolism and dyslipidaemia in Alstrom syndrome will be discussed as well as genotypic and phenotypic variation in the syndrome in the context of recent metabolic studies in insulin resistance. Potential therapies including low-carbohydrate diet, niaspan and incretins will be considered. SUMMARY: These new insights will encourage early introduction of protective nutrition, exercise and drug therapies in Alstrom syndrome and suggest novel approaches to understanding diabetes mellitus, hyperlipidaemias and heart disease in the general population. PMID- 19550325 TI - Phospholipase A2 inhibitors. AB - PURPOSE OF REVIEW: As the role of lipids and inflammation in the genesis and progression of the atherosclerosis disease is unquestionable, novel treatment modalities that target both aspects are currently under investigation. RECENT FINDINGS: For a long time atherosclerosis was regarded as a lipid-driven disease, but now it is evident that it also involves the simultaneous and combined effect of inflammation and immunological pathways. The secreted PLA2s and the lipoprotein-associated phospholipase A2 (Lp-PLA2) have been associated with atherogenesis and its complications. These two enzymes produce biologically active metabolites that are involved in several phases of the atherosclerosis process. SUMMARY: In animal, pathological and epidemiological studies, the increased levels of these two phospholipases (i.e. PLA2s and Lp-PLA2) have been related with an increase in complex coronary lesions and increase in major cardiovascular clinical events, respectively. Therefore, inhibition of these enzymes has become the focus of research in this last decennium. Novel pharmacological inhibitors of those enzymes such as darapladib and varespladib emerge as promising therapeutical options for treating patients with coronary artery disease. Ongoing mechanistic and clinical outcome trials will further elucidate their role in this context. PMID- 19550326 TI - The management of intrauterine synechiae. AB - PURPOSE OF REVIEW: Intrauterine adhesions are a rare but significant cause of menstrual disturbance and infertility. Most cases are caused by uterine instrumentation. It is important for clinicians to understand the cause, diagnostic tools and rationale behind treatment. RECENT FINDINGS: Hysteroscopy is the gold standard for diagnosis and treatment of intrauterine adhesions. A combination of blunt and sharp dissection or electrosurgery is used by most units. Antibiotics and postoperative administration of estrogen +/- progestogen is important in prevention of recurrence. The use of intrauterine contraceptive devices following synechiolysis is supported by some groups. Restoration of menstruation is highly successful (more than 90%), and pregnancy rates around 50 60% with live birth rates around 40-50% can be achieved. SUMMARY: Clinicians should maintain a level of suspicion of intrauterine adhesions and should investigate by hysteroscopy if necessary. Treatment should follow a protocol that incorporates sound hysteroscopic technique with antibiotic prophylaxis and postoperative hormonal therapy. Consideration should be given to the use of an intrauterine contraceptive device or Foley catheter for a short period. PMID- 19550327 TI - Insulin resistance in women's health: why it matters and how to identify it. AB - PURPOSE OF REVIEW: To examine the significance of insulin resistance in women's health and review methods for diagnosing it. RECENT FINDINGS: Clinical phenotypes in conjunction with standard clinical biochemical assays, that is, the metabolic syndrome, remain the key method to diagnose insulin resistance in clinical practice. Candidate alleles from type 2 diabetes offer little predictive value for cardiovascular events beyond traditional risk factors. Simple environmental factors such as irregular meal frequency appear to increase the risk of the metabolic syndrome and require greater scrutiny. Pregnancy complications, particularly gestational diabetes and preeclampsia in the mother and preterm birth in the fetus are events that suggest elevated risk for future cardiovascular morbidity in those affected. SUMMARY: Clinical phenotypes of insulin resistance identify women at risk for perinatal and reproductive complications. PMID- 19550328 TI - IgG4-related interstitial lung disease: a new and evolving concept. AB - PURPOSE OF REVIEW: This review examines recent advances in our knowledge of the clinical, pathological, diagnostic, and therapeutic aspects of IgG4-related interstitial lung disease (ILD). RECENT FINDINGS: A recent case series of ILD with IgG4-positive plasma cells suggested grade 1 lymphomatoid granulomatosis. The presence of the IgG4-positive plasma cells with the lack of atypical cells favored IgG4-related ILD as a diagnosis. In another case study, four out of 30 patients with autoimmune pancreatitis developed pulmonary involvement during follow-up. Elevations of IgG4 and Krebs von den Lungen-6 levels were associated and thought to be predictive of the development of IgG4-related lung disease. A retrospective analysis investigating radiological/pathologic correlation in IgG4 lung disease identified computed tomographic features pathologically corresponding to IgG4-related sclerosing inflammation in the pulmonary interstitium. SUMMARY: IgG4-related ILD is a new and evolving entity. It can occur with or without systemic involvement. Larger studies are necessary to elucidate the exact mechanism and clinical characteristics of this disorder. PMID- 19550329 TI - Update in idiopathic pulmonary fibrosis. AB - PURPOSE OF REVIEW: This review aims to highlight and place in context recent advances in and insights into the natural history, diagnosis, and management of idiopathic pulmonary fibrosis (IPF). RECENT FINDINGS: Although the diagnosis of IPF remains challenging, an evolution in systems of practice and advancing technologies are steadily improving diagnostic accuracy. The identification of concomitant pulmonary hypertension as well as acute exacerbations of the underlying disease have taken on increasing importance in the natural history of IPF. Similarly, the management of IPF remains challenging, and although a number of recent trials of novel investigational agents for the treatment of IPF yielded negative results, at least one of these trials showed significant benefit suggesting progress in the treatment of this disease. SUMMARY: Although IPF remains a diagnostic and therapeutic challenge to even the most experienced of clinicians, our knowledge of the natural history of the disease, diagnostic accuracy, and therapeutic approach continue to advance. PMID- 19550331 TI - Risk factors for functional decline in osteoarthritis of the hip or knee. AB - PURPOSE OF REVIEW: To summarize recent studies on risk factors for functional decline (i.e. worsening of pain and activity limitations) in osteoarthritis of the hip or knee; after a long period of neglect, information on risk factors for functional decline has markedly improved in recent years. RECENT FINDINGS: Studies show slow worsening of pain and activity limitations over time. There is a considerable between-individual variation in the course of pain and activity limitations; identification of risk factors for functional decline is therefore highly relevant. Risk factors for functional decline include characteristic physical impairments in osteoarthritis (pain, stiffness, reduced muscle strength, laxity of the knee joint, proprioceptive inaccuracy, poor standing balance and impaired range of joint motion), cognitive and visual impairments, comorbidity and overweight, psychological and social factors (anxiety, depression, fatigue, poor self-efficacy and social support), health behaviors and sociodemographic factors. SUMMARY: The information on risk factors can be used pragmatically to construct prediction rules on functional decline in osteoarthritis patients. Both clinicians and patients value the ability to prognosticate future pain and activity limitations. Furthermore, the information on risk factors can be used to construct causative models to explain pain and activity limitations: examples include 'instability of joints' and 'avoidance of activity' as causal mechanism of pain and activity limitations. Improved understanding of pain and activity limitations is the basis for improved treatment and rehabilitation of osteoarthritis patients. PMID- 19550333 TI - Determining prognosis and predicting survival in end-of-life care. AB - PURPOSE OF REVIEW: The purpose of this article is to review some recent studies referred to prognosis factors and our capacity to predict death during the final stages of disease. RECENT FINDINGS: Recent studies show significant approaches through the use of some instruments (especially self-assessment tools), which we will analyze herein. SUMMARY: In many cases of clinical practice, this prediction capacity based on solid elements may be key to determine or contraindicate certain therapeutic resources. Such is the case, for example, when it is necessary to define the transfer of a patient to a proper specialized center, under family or legal circumstances demanding a clarification on this point, or for handling the dialog with the patients and/or their relatives. According to recent findings, we should include some elements of self-assessment in our instruments to predict death in palliative care. PMID- 19550332 TI - Advances in the treatment of neutropenia. AB - PURPOSE OF REVIEW: The present review updates treatment of neutropenia from articles published from January 2008 through April 2009. RECENT FINDINGS: Chemotherapy-induced neutropenia occurs most commonly in the first cycle of treatment. Older patients, patients with multiple comorbidities, and those receiving more myelotoxic drugs are prone to develop neutropenia and its complications. Current guidelines recommend the prophylactic use of the myeloid growth factors for the first cycle of chemotherapy for patients with more than a 20% risk of febrile neutropenia. Meta analysis from randomized trials shows that granulocyte colony-stimulating factor prophylaxis is associated with patients receiving more intensive chemotherapy, having better survival, but also having a higher risk of secondary acute myeloid leukemia. Antibiotics are standard treatment of febrile neutropenia and are increasingly used for prophylaxis in 'low-risk' patients. SUMMARY: The myeloid growth factor granulocyte colony stimulating factor has radically changed our approach to the prevention of febrile neutropenia. Antibiotics remain the mainstay of treatment of febrile neutropenia. PMID- 19550330 TI - T-cell and B-cell signaling biomarkers and treatment targets in lupus. AB - PURPOSE OF REVIEW: Systemic lupus erythematosus is characterized by the production of antinuclear autoantibodies and dysfunction of T-cells, B-cells, and dendritic cells. Here, we review newly recognized genetic factors and mechanisms that underlie abnormal intracellular signal processing and intercellular communication within the immune system in systemic lupus erythematosus. RECENT FINDINGS: Activation of the mammalian target of rapamycin plays a pivotal role in abnormal activation of T and B-cells in systemic lupus erythematosus. In T-cells, increased production of nitric oxide and mitochondrial hyperpolarization were identified as metabolic checkpoints upstream of mammalian target of rapamycin activation. Mammalian target of rapamycin controls the expression T-cell receptor associated signaling proteins CD4 and CD3zeta through increased expression of the endosome recycling regulator HRES-1/Rab4 gene, mediates enhanced Ca2+ fluxing and skews the expression of tyrosine kinases both in T and B-cells, and blocks the expression of Foxp3 and the expansion of regulatory T-cells. Mitochondrial hyperpolarization and the resultant ATP depletion predispose T-cells to necrosis, thus promoting the dendritic cell activation, antinuclear autoantibody production, and inflammation. SUMMARY: Mitochondrial hyperpolarization, increased activity of mammalian target of rapamycin and Syk kinases, enhanced receptor recycling and Ca2+ flux have emerged as common T and B-cell biomarkers and targets for treatment in systemic lupus erythematosus. PMID- 19550334 TI - Second-line therapy in bladder cancer. AB - PURPOSE OF REVIEW: To review new developments in second-line treatment for transitional cell carcinoma of the urinary bladder focusing on advances and findings within the last year. RECENT FINDINGS: So far no standard therapy has been established for pretreated patients with transitional cell carcinoma. Prognostic and predictive factors for response and outcome in pretreated patients have been studied retrospectively and will help to identify those who might benefit from intensive therapy and those who would rather improve their quality of life with best supportive care. Single-agent chemotherapy in this setting provided low response rates, short progression-free and overall survival. Vinflunine, a novel vinca-alkaloid, and best supportive care had a modest, but still significant benefit at a very moderate toxicity rate in a randomized phase III trial. As for second-line combination chemotherapy, response rates as well as potential toxicities are higher and the progression-free survival and overall survival benefits are still poorly understood. New chemotherapeutics and approaches like metronomic chemotherapy and novel agents are under investigation. SUMMARY: With vinflunine there is reasonable hope for a new standard chemotherapy in second-line management of transitional cell carcinoma. However, due to the reported results with single agents and combination chemotherapy, treatment within clinical trials is still the best choice in patients progressing after first-line chemotherapy. PMID- 19550335 TI - The oncologic results of laparoscopic radical cystectomy are not (yet) equivalent to open cystectomy. AB - PURPOSE OF REVIEW: To compare oncologic outcomes in a contemporary series of patients undergoing radical cystectomy (RCX) by the laparoscopic or open approach. RECENT FINDINGS: Laparoscopic RCX with extracorporeally constructed urinary diversion is a safe and effective operation for appropriate patients with bladder cancer. Perioperative and functional outcomes are comparable with open surgery. Worldwide experience continues to increase; more than 1000 surgeries have already been performed. Intermediate-term oncologic outcomes appear to be comparable to open approach. SUMMARY: When stage distribution (organ-confined versus nonorgan-confined) and survival rates of laparoscopic and open RCX are cross-checked against surrogate markers (LN+, margin+, distant failure, local recurrence rate, etc.), it becomes easily obvious that the laparoscopic RCX and open cohorts are not identical. To explain the observed discrepancy, there must be a strong selection bias. Data on overall, disease-specific and recurrence-free survival on laparoscopic RCX are still immature compared with the standard of care that must remain in open RCX. To prove the noninferiority of laparoscopic compared with open surgery regarding outcome, multicenter prospective trials are urgently needed. Until then laparoscopic RCX must be considered experimental surgery. PMID- 19550336 TI - Effects of contralateral noise on 40-Hz and 80-Hz auditory steady-state responses. AB - OBJECTIVES: The amplitude of 40-Hz auditory steady-state response (ASSR) in response to repeated click or tone pips at levels of 45 to 60 dB SL is reduced by up to 50% by the central masking mechanism if white noise at levels of 40 to 60 dB SL is applied contralaterally. However, the effect of contralateral noise on the elevation of the threshold of ASSR is unknown. The present study investigated the effects of contralateral noise on the threshold measurements of ASSR for 40- and 80-Hz amplitude modulated tones that are widely used in clinical examinations. DESIGN: The effects of contralateral noise on the 40- and 80-Hz ASSRs for amplitude modulated tones at 500 Hz and 2000 Hz were examined in 11 healthy volunteers (10 men and 1 woman, mean age 26.1 years). Contralateral noise consisted of white noise low-pass filtered at 700 and 4000 Hz in the measurements of ASSR at a carrier frequency of 500 and 2000 Hz, respectively. RESULTS: Contralateral noise at a level of 40 dB SL caused no significant psychophysical threshold elevation, caused significant threshold elevation of the 40-Hz ASSR (average 10 to 15 dB), and caused no significant threshold elevation of the 80-Hz ASSR. CONCLUSIONS: The different effects on the 40- and 80-Hz ASSRs were probably related to the differences in sources because the 40-Hz ASSR contains more components from the upper auditory pathway that are affected by contralateral masking, whereas the 80-Hz ASSR contains more components from the brain stem. The present results suggest that threshold elevation of the 40-Hz ASSR may occur during clinical measurements using binaural presentation of sounds and cause a possible discrepancy between psychophysical threshold and 40-Hz ASSR measurements. PMID- 19550337 TI - The effectiveness of dexmedetomidine infusion for sedating oral cancer patients undergoing awake fibreoptic nasal intubation. AB - BACKGROUND AND OBJECTIVE: Dexmedetomidine is characterized with effects of sedation, analgesia, amnesia and lack of respiratory depression. Hence, it should be suitable for awake fibreoptic intubation (AFOI). METHODS: We enrolled 30 oral cancer patients with limited mouth openings who were undergoing AFOI for elective surgery. Patients were randomly allocated into two groups; the Dex group (n = 16) that received dexmedetomidine (1.0 microg kg(-1)) infusion and the Control group (n = 14) that received fentanyl (1.0 microg kg(-1)) infusion. Main outcomes were evaluated by grading scores presenting conditions for nasal intubation and postintubation. Other analysed parameters included airway obstruction, haemodynamic changes, consumption time for intubation, amnesia level and satisfaction. RESULTS: Intubation score (1-5) representing condition for nasal intubation was significantly better in the Dex group [2(1-3)] than in the Control group [3(2-5)] (P = 0.001). Postintubation score (1-3) representing tolerance to intubation also showed more favourable results in the Dex group [1(1-3)] than in the Control group [2(2-3)] (P = 0.002). The Dex group showed significantly reduced haemodynamic response to intubation than the Control group. Incidence requiring temporary haemodynamic support was higher in the Dex group but not of significance. Both levels of amnesia and satisfaction score were significant in the Dex group. Other analysed parameters such as consumption time for intubation, airway obstruction score and postoperative adverse events did not differ significantly. CONCLUSION: Combination of dexmedetomidine loading with topical anaesthesia provides significant benefit for AFOI in intubation condition, patient tolerance, haemodynamic response, amnesia and satisfaction. Dexmedetomidine is effective for AFOI in anticipated difficult airway with only minor and temporary haemodynamic adverse effects. PMID- 19550338 TI - The influence of perioperative oxygen concentration on postoperative lung function in moderately obese adults. AB - BACKGROUND AND OBJECTIVE: Obesity aggravates the negative effects of general anaesthesia and surgery on the respiratory system, resulting in decreased functional residual capacity and expiratory reserve volume, and increased atelectasis and ventilation/perfusion (Va/Q) mismatch. High-inspired oxygen concentrations also promote atelectasis. This study compares the effects of perioperative inspired low-oxygen and high-oxygen concentrations on postoperative lung function and pulse oximetry values in moderately obese patients (BMI 25-35). METHODS: We prospectively studied 142 overweight patients, BMI 25-35, undergoing minor peripheral surgery; they were randomly allocated to receive either low inspired or high-inspired oxygen concentrations during general anaesthesia. Premedication, general anaesthesia and respiratory patterns were standardized. Arterial oxygen saturation (pulse oximetry) was measured on air breathing. Inspiratory and expiratory lung functions were measured preoperatively (baseline) and at 10 min, 0.5, 2 and 24 h after extubation with the patient supine, in a 30 degrees head-up position. The two groups were compared using repeated-measure analysis of variance and t-test analysis. RESULTS: The low-inspired oxygen group had significantly better arterial saturation during the first 24 h (P < 0.01). Mid-expiratory flow 25 values indicating small airway collapse were significantly better in the low-oxygen group at all measurements (P < 0.05). CONCLUSION: We conclude that postoperative lung function and arterial saturation is better preserved by a low-oxygen strategy, although it is not clear whether this has clinical relevance for the prevention of postoperative pulmonary complications. PMID- 19550339 TI - Dedicated endotracheal tube for percutaneous tracheostomy. AB - BACKGROUND AND OBJECTIVE: Percutaneous dilational tracheostomy (PDT) is an accepted method in intensive care patients. In our neurosurgical ICU, we routinely perform PDT. We have performed 130 PDT procedures and have often observed previously reported difficulties/complications: dislocation of the endotracheal tube (ETT); frequent puncture of the ETT cuff; difficult visualization of the entire procedure because the ETT is often in the way; and risk of damaging the posterior tracheal wall due to needle puncture or rotating dilator. We believe that the rate of complications is directly related to the ability and experience of the anaesthetist carrying out the PDT. Because our hospital is a university teaching centre, PDT is often carried out by residents who are not experienced with the technique. In order to guarantee the patient's safety and to optimize the learning curve, we propose, in such circumstances, the use of a modified standard ETT. METHOD: We developed a prototype tube for percutaneous tracheostomy procedures modifying a commercially available ETT. The idea was to create a protection shield for the tracheal wall, greater stability of the ETT in the trachea and improved vision during the whole procedure. We tested the effectiveness of the prototype on a manikin model. CONCLUSION: The dedicated ETT eliminates puncturing of the ETT, rupture of the cuff and difficult direct vision. We report an optimal stability of the tube during the whole procedure and improvement in the efficacy and safety of the procedure. When residents attempt PDF for the first time, we believe that it would be safer and more practical to perform such a technique with the dedicated ETT. PMID- 19550340 TI - The effect of dexmedetomidine sedation on brachial plexus block in patients with end-stage renal disease. AB - BACKGROUND AND OBJECTIVE: The study investigated the effect of intravenous dexmedetomidine on brachial plexus block in patients with end-stage renal disease sedated with dexmedetomidine or midazolam for the formation of arteriovenous fistula. METHODS: A supraclavicular brachial plexus block was performed in 68 patients scheduled for an arteriovenous fistula formation. A successful block was achieved in 64 patients (94%). Sixty-four patients received either dexmedetomidine (loading dose 1 microg kg 10 min followed by infusion of 0.2-0.7 microg kg h) or midazolam (bolus dose 0.04 mg kg followed by infusion of 0.04 0.08 mg kg h). Three patients were excluded during the study. Finally, 61 patients were analysed, 32 in the dexmedetomidine group and 29 in the midazolam group. Administration of the study drug was started after achievement of a complete block. The duration of the motor and sensory block was assessed. RESULTS: The motor and sensory block was longer in the dexmedetomidine group (11.9 +/- 3.8 vs. 9.4 +/- 3.4 h, P = 0.0085 and 9.4 +/- 3.4 vs. 7.3 +/- 2.8 h, P = 0.030, respectively). CONCLUSION: A systemic administration of dexmedetomidine prolongs the duration of brachial plexus block. PMID- 19550341 TI - Effective anti-tumor responses induced by recombinant bacillus Calmette-Guerin vaccines based on different tandem repeats of MUC1 and GM-CSF. AB - In this study, we constructed several novel breast cancer vaccines, Bacillus Calmette-Guerin (BCG)-MUC1 variable-number tandem repeats (VNTR) 1/4/8-CSF, that consist of BCG and express 1, 4, and 8 of the tandem repeats of MUC1 and human granulocyte-macrophage colony-stimulating factor (GM-CSF). The ability of the three recombinant BCG vaccines to inhibit breast cancer growth was observed in human (hu)-peripheral blood lymphocyte (PBL)-severe combined immunodeficient (SCID) mice. Prophylactic protective responses were successfully induced and the tumor incidence in mice immunized with recombinant BCG (rBCG)-MVNTR4-CSF (37.5%) or rBCG-MVNTR8-CSF (25%) was significantly decreased compared with control (100%) at 42 days after tumor implantation (P<0.05 vs. control group). We also found that CD4-positive and CD8-positive lymphocytes were detected only in tumors grown in rBCG-MVNTR4/8-CSF-immunized animals, and strong IFN-gamma responses were induced by immunization with rBCG-MVNTR4-CSF and rBCG-MVNTR8-CSF vaccines. This study suggests a potential role of coexpressed GM-CSF and tandem repeats of MUC1 in eliciting tumor-specific immune response. Our results indicate that rBCG MVNTR4-CSF and rBCG-MVNTR8-CSF immunization preventively inhibited breast cancer growth in hu-PBL-SCID mice and the both rBCG vaccines may be good candidates for breast cancer immunotherapy. PMID- 19550342 TI - Female representation on emergency medicine editorial teams. AB - OBJECTIVE: To analyse the presence of women on the editorial teams of emergency medicine journals and the potential relationship between the pre-eminence of the journal and their presence. MATERIALS AND METHODS: In this cross-sectional study, we examined 10 journals cited under the heading of 'Emergency Medicine' by Thomson Scientific in the Journal Citation Reports and 14 additional emergency journals not cited but which publish investigations in emergency medicine. We evaluated the editorial board posted on their websites, determining the number of men and women occupying executive tasks, as well as the sex of the editor-in chief of each journal. RESULTS: We identified 372 people working on the editorial teams (mean: 15.5, SD: 13.5), 49 being women (13.2%). Of these 372 people, 28 were editors-in-chief but only one was female (3.6%). We found no statistical differences between indexed and nonindexed emergency journals regarding female representation on the editorial team or in the position of editor-in-chief. Neither did we find any relationship between female presence and the pre-eminence of the indexed journals using impact factor as a surrogate marker. CONCLUSION: Very few women are found either on editorial teams or in editor-in-chief positions in the emergency medicine journals, irrespective of the pre-eminence of the journal. It should be investigated whether a negative journal bias underlies these findings. PMID- 19550343 TI - Prehospital stroke scales in a Belgian prehospital setting: a pilot study. AB - OBJECTIVE: To compare the diagnostic value of the Cincinnati Prehospital Stroke Scale, the Face Arm Speech Test, the Los Angeles Prehospital Stroke Screen and the Melbourne Ambulance Stroke Screen for identifying patients with an acute stroke in a prehospital setting in Belgium. METHODS: A prospective study was performed, using a questionnaire for every primarily transported patient within emergency medial service with relevant neurological complaints. Exclusion criteria were: patients below 18 years, trauma victims, Glasgow Coma Scale of less than 8 or transported to another hospital. The questionnaire is a comprehension of different stroke scales. RESULTS: The Face Arm Speech Test and Cincinnati Prehospital Stroke Scale demonstrate a high sensitivity (95%) but a lower specificity (33%). The sensitivity of the Los Angeles Prehospital Stroke Screen and Melbourne Ambulance Stroke Screen was lower (74%), but the specificity increased (83 and 67%). Items investigating unilateral facial paralysis and unilateral loss/absence of motor response in upper extremities seemed to be most discriminating between the stroke group (68-78%) and the nonstroke group (17%), suggesting that items related to clinical assessment are more important in stroke recognition than history items. Combination of all clinical parameters of the different scores resulted in a sensitivity and specificity of 95 and 83%, respectively. CONCLUSION: The results obtained in this study are comparable with earlier investigations. Given the limitations of the study, we could not identify the most adequate stroke scale. History items seem to be less relevant compared with clinical assessment. Further research is needed to determine the most adequate stroke scale. PMID- 19550344 TI - Monitoring of therapy in patients with chronic hepatitis B virus. AB - OBJECTIVE: The aims of this study were to evaluate therapy with lamivudine (LAM) and adefovir dipivoxil (ADV) monotherapy in chronic hepatitis B virus (HBV) infected patients with frequent measurements of DNA levels, to characterize HBV genotypes, and to determine the emergence of nucleos(t)ide analogue mutants before and during the therapy by direct-sequencing the reverse transcriptase region and by INNO-LiPA HBV DR v3. MATERIALS AND METHODS: A total of 15 chronic HBV patients were analysed: 11 were treated with ADV and four were treated with LAM. RESULTS: Viral genotype was determined, showing the presence of genotype D (73%) in 11 patients and genotype A (27%) in four patients. In the viral response to treatment, three patients developed substitutions at rtM204I associated with LAM resistance and one of these patients presented rtM204V/I plus rtL180M mutation. In contrast, of the 11 patients treated with ADV, three patients developed mutations (rtN236T; rtA181V; rtA181V plus rtN236T). With regard to this case, the same results were observed by INNO-LiPA HBV DR v3 and direct sequencing, but by direct sequencing we detected an extra mutation rtQ215S that was present in two patients: one patient who was on treatment with LAM had an rtQ215S mutation in addition to an rtM204I, and the second patient treated with ADV had rtA181V. CONCLUSION: Direct sequence analysis is an essential tool to optimize therapeutic management of HBV chronic infection in clinical practice to choose the appropriate nucleos(t)ide analogues and to detect extra mutations that are not included in the commercial kit. PMID- 19550345 TI - Case report: appearance of an intestinal metastasis from intrahepatic cholangiocarcinoma occurring 5 years after resection of the primary tumor. AB - We present the unusual case of a 76-year-old male who developed an intestinal recurrence of the same tumor 5 years after hepatic resection for an intrahepatic cholangiocarcinoma. At the time of the first surgery, the patient had undergone hepatic bisegmentectomy of segments IV and V with an 'en bloc' gallbladder resection and porta hepatis lymphadenectomy for the presence of a focal cholangiocarcinoma measuring about 3.0 x 2.5 cm in diameter. The histological report confirmed intrahepatic cholangiocarcinoma, the resection margins were free from disease, and there were no lymph node metastases. Five years later colonoscopy showed, at the level of the splenic flexure, the presence of a sessile bilobate polypoid neoplasm. The patient underwent left hemicolectomy with a histological diagnosis of an isolated recurrence of cholangiocarcinoma. PMID- 19550346 TI - Hepatocellular carcinoma in two patients with cardiac cirrhosis. AB - Hepatocellular carcinoma has been commonly associated with multiple etiologic factors including hepatitis B and C, alcoholic liver disease, and more rarely congenital metabolic liver diseases. 'Cardiac cirrhosis' is the cirrhosis resulting from prolonged passive liver venous congestion secondary to right-sided congestive heart failure; hepatocellular carcinoma is a rarely reported outcome. In this study we present two female patients with congenital heart defects treated with the Fontan procedure who survived into their third decade, and developed hepatocellular carcinoma in the setting of cardiac cirrhosis. The Fontan procedure diverts blood from the inferior vena cava and superior vena cava to the pulmonary arteries, thereby increasing survival in infants born with a single effective ventricle. As such patients live longer, however elevated pulmonary and right-sided heart pressures cause chronic passive liver congestion and eventual cardiac cirrhosis. The two patients in this study had no risk factors for hepatocellular carcinoma other than cardiac cirrhosis secondary to their prolonged survival after their Fontan procedure. In conclusion, we suggest that cardiac cirrhosis may be a risk factor for developing hepatocellular carcinoma and recommend close follow-up and hepatocellular carcinoma screening for patients with known right heart failure and passive hepatic congestion. PMID- 19550348 TI - Sex differences in dietary coping with gastrointestinal symptoms. AB - AIM: Nutritional changes are often considered first-line treatment in public health diseases that apply to many gastrointestinal (GI) disorders, as different food and beverages may modulate GI motor and sensory functions, and may provoke GI symptoms. The aim of this study was to examine dietary coping and possible changes in food and beverage intake in relation to GI symptoms reported by identified irritable bowel syndrome (IBS) patients compared with healthy controls, and whether any sex differences were observed in these respects. METHODS: A population-based case-control design was used. Three primary healthcare centres were selected in the city of Linkoping in Sweden. The IBS patients were recruited from the studied primary healthcare centers on the basis of diagnoses from computerized medical records. The controls were randomly selected from the general population in the same region. A questionnaire was used with specific questions about self-reported food and beverage increase or decrease of GI symptoms and self-reported changes in dietary habits. RESULTS: Female IBS patients seem to be more willing to change dietary habits because of their GI problems than men. Effects of these nutritional behaviour changes were reported for almost all participants that had made dietary adjustments. Fatty food, certain vegetables, dairy products and eggs were significantly more reported to cause GI complaints among IBS patients compared with their controls. CONCLUSION: Female IBS patients reported more changes in their dietary habits because of GI problems than men with the disease. The majority of both women and men who changed their dietary habits because of GI problems experienced improvement in their symptoms. PMID- 19550347 TI - Economic evaluation of early monotherapy versus delayed monotherapy or combination therapy in patients with acute hepatitis C in Germany. AB - BACKGROUND: Antiviral treatment of acute hepatitis C virus (HCV) almost doubles the chance of sustained virological response (SVR) compared with that achievable by treating chronic HCV. AIM: To conduct a health economic evaluation comparing early and delayed therapies for acute HCV in Germany. METHODS: One hundred and thirty-three patients with acute HCV were evaluated in two early monotherapy (EMT) studies and 60 in a delayed therapy study. Efficacy was determined by SVR. In the EMT studies, patients were treated with either standard or pegylated interferon for 24 weeks. In the delayed therapy study, patients with persisting infection were treated with interferon monotherapy or combination therapy with ribavirin for a median of 36 weeks. We conducted a cost-effectiveness analysis based on the study results and a linear simulation model based on current treatment recommendations. RESULTS: The SVR rate for the sex-adjusted on treatment analysis between early and delayed therapies was not significantly different (92.7 vs. 90.9%; P = 0.7). Medication costs accounted for more than 90% in both treatment options. Direct medical costs of early therapy (euro7064/patient) were euro321 lower than those of delayed therapy (P = 0.8). The incremental cost-effectiveness ratio was -178 euro/SVR(%) (confidence interval: -224 to 360 euro/SVR(%)). Average modeled direct medical costs of delayed combination therapy were from euro6745 to euro8299 per patient (from approximately 7% less up to 15% higher than EMT). Spontaneous viral clearance and therapy duration were the most sensitive variables. CONCLUSION: There was no significant efficacy and cost difference between therapy alternatives in base cases. However, in the majority of scenarios in the sensitivity analyses, EMT was a more cost-effective option in acute HCV therapy. PMID- 19550349 TI - An algorithm to optimize viral load testing in HIV-positive patients with suspected first-line antiretroviral therapy failure in Cambodia. AB - OBJECTIVE: To develop an algorithm for optimal use of viral load testing in patients with suspected first-line antiretroviral treatment (ART) failure. METHODS: Data from a cohort of patients on first-line ART in Cambodia were analyzed in a cross-sectional way to detect markers for treatment failure. Markers with an adjusted likelihood ratio <0.67 or >1.5 were retained to calculate a predictor score. The accuracy of a 2-step algorithm based on this score followed by targeted viral load testing was compared with World Health Organization criteria for suspected treatment failure. RESULTS: One thousand eight hundred three viral load measurements of 764 patients were available for analysis. Prior ART exposure, CD4 count below baseline, 25% and 50% drop from peak CD4 count, hemoglobin drop of > or =1 g/dL, CD4 count <100 cells per microliter after 12 months of treatment, new onset of papular pruritic eruption, and visual analog scale <95% were included in the predictor score. A score >or=2 had the best combination of sensitivity and specificity and required confirmatory viral load testing for only 9% of patients. World Health Organization criteria had a similar sensitivity but a lower specificity and required viral load testing for 24.9% of patients. CONCLUSION: An algorithm combining a predictor score with targeted viral load testing in patients with an intermediate probability of failure optimizes the use of scarce resources. PMID- 19550350 TI - Loss to follow-up of adults in public HIV care systems in central Mozambique: identifying obstacles to treatment. AB - INTRODUCTION: Access to antiretroviral treatment (ART) has expanded dramatically in resource-limited settings. Evaluating loss to follow-up from HIV testing through post-ART care can help identify obstacles to care. METHODS: Routine data were analyzed for adults receiving services in 2 public HIV care systems in central Mozambique. The proportion of people passing through the following steps was determined: (1) HIV testing, (2) enrollment at an ART clinic, (3) CD4 testing, (4) starting ART if eligible, and (5) adhering to ART. RESULTS: During the 12-month study period (2004-2005), an estimated 23,430 adults were tested for HIV and 7005 (29.9%) were HIV positive. Only 3956 (56.5%) of those HIV positive enrolled at an ART clinic < or =30 days after testing. CD4 testing was obtained in 77.1% < or =30 days of enrollment. Of 1506 eligible for ART, 471 (31.3%) started ART < or =90 days after CD4 testing. Of 382 with > or =180 days of potential follow-up time on ART, 317 (83.0%) had pharmacy-based adherence rates > or =90%. DISCUSSION: Substantial drop-offs were observed for each step between HIV testing and treatment but were highest for referral from HIV testing to treatment sites and for starting ART. Interventions are needed to improve follow up and ensure that people benefit from available HIV services. PMID- 19550351 TI - Lifetime risk factors for HIV/sexually transmitted infections among male-to female transgender persons. AB - OBJECTIVES: To describe and evaluate risk factors for HIV/sexually transmitted infections (STIs) among male-to-female (MTF) transgender persons. METHODS: Using the life chart interview, potential lifetime risk factors for HIV/STIs among MTFs were measured and evaluated in conjunction with lifetime exposures for HIV, syphilis, hepatitis B, and hepatitis C. The participants were 517 MTFs between the ages of 19 and 59 years from the New York metropolitan area. RESULTS: HIV/STIs were low among white Americans and very high among Hispanics and African Americans. In the latter groups, HIV and hepatitis B were associated with an androphilic sexual orientation, lifetime number of commercial sex partners (sex work), and the social expression of transgender identity; syphilis was associated with lifetime number of casual sex partners; and hepatitis C was associated with injection drug use, unemployment, and social expression of transgender identity. In multivariate models, the social expression of transgender identity was the strongest and most consistent predictor of HIV/STIs. Consistent with their lower levels of infections, white Americans reported significantly lower levels of the risk factors found to be predictive of HIV/STI among Hispanics and African Americans. CONCLUSIONS: HIV/STI prevention in this population should be targeted at Hispanic and African Americans. Prevention programs should incorporate multiple components designed to address the diverse issues confronting ethnic minority transgender persons, with an emphasis on the social expression of transgender identity. PMID- 19550352 TI - Computed tomography coronary angiography plaque burden in patients with suspected coronary artery disease. AB - PURPOSE: To determine the relationship between established cardiovascular risk factors, clinical presentation and the extent of coronary artery disease (CAD), as described with computed tomography coronary angiography. MATERIAL AND METHODS: In this cross-sectional study, we included 567 symptomatic individuals without a history of CAD who consecutively underwent 64-slice computed tomography coronary angiography for evaluation of suspected CAD. We analyzed the prevalence of CAD depending on sex, age, symptoms and risk factors. RESULTS: A total of 8542 segments were analyzed. No evidence of CAD was observed in 225 patients (40%), nonsignificant CAD in 221 patients (39%) and significant CAD (luminal narrowing >50%) in the remaining 121 patients (21%). CAD increased with advancing age, significantly above 50 years (P < 0.05). Female patients had a higher prevalence of normal coronary arteries and males of significant CAD (P < 0.01). With the increase of risk factors, there was a significant increase of the significant disease (P < 0.01). Typical pain with respect to atypical pain had the strongest association with significant CAD (16 vs. 38%; P < 0.05). In multivariate analysis, the number of risk factors, age, male sex and typical pain remained strong predictors of significant CAD (P < 0.0001). CONCLUSION: Computed tomography coronary angiography may play an important role in risk stratification of patients with suspected CAD. PMID- 19550353 TI - Thin strut chrome-cobalt stent implantation for treatment of de-novo lesions in small coronary vessels: results of the RISICO Italian Registry (Registro Italiano Mini VISION nei piccolo Vasi) utilizing the Mini VISION coronary stent platform. AB - OBJECTIVES: The Registro Italiano Mini VISION nei piccoli Vasi registry is a prospective, multicenter, observational study, aimed at assessing immediate and long-term angiographic and clinical outcomes of a small-vessel cobalt-chrome super alloy-dedicated stent (Multi-Link RX VISION) in de-novo and long lesions. BACKGROUND: Small artery size is an important determinant of poor outcomes in percutaneous coronary interventions. METHODS: Patients with ischemic heart disease were included. The primary end point was procedural success. Secondary end points included clinical restenosis (need for target lesion revascularization ), incidence of major adverse cardiac events at 6 months, and cost-effectiveness analysis. RESULTS: Between September 2004 and October 2005, 143 patients (mean age 67 +/- 11 years; 22% diabetes) were enrolled; 6-month follow-up was completed in May 2006. Average lesion length, mean stent length and diameter were 16.8 +/- 7.1, 17.01 +/- 3.9 and 2.41 +/- 0.14 mm, respectively. Procedural success was 96%. At 6-month follow-up, the hierarchical major adverse cardiac event rate was 11.6%, 2.9% deaths, 2.9% myocardial infarction and 5.8% target lesion revascularization. Cost-effectiveness analysis will be reported in a further publication. CONCLUSION: Small-vessel disease treatment with Mini VISION stents permits an elevated procedural success rate with low incidence of clinical restenosis and major adverse cardiac events at mid-term follow-up. Such results require confirmation by means of a randomized controlled study against drug eluting stents. PMID- 19550354 TI - Edge-to-edge technique: is it also useful in children? AB - OBJECTIVE: Mitral valve prolapse is a rare condition and represents a challenge for cardiac surgeons. Many techniques have been described to treat the wide spectrum of abnormalities causing mitral valve regurgitation but the low incidence of these abnormalities is the cause of the relatively poor experience of many institutions. METHODS: From April 2005 to September 2006, five patients underwent isolated mitral valve repair with the edge-to-edge technique for anterior or bileaflet prolapse. Three patients also had a Gore-Tex strip to reinforce the posterior annulus, whereas two had a 26-size soft incomplete ring annuloplasty. RESULTS: There was no death and none of the patients required reoperation. None of the patients had mitral regurgitation more than 1+ at a mean follow-up of 30 months. None of the patients had mitral stenosis or any valve related event. CONCLUSION: The edge-to-edge technique is a well tolerated and useful technique for the anterior or bileaflet prolapse of the mitral valve. Although many techniques have been described with good results, in many anatomical settings and in small patients, edge-to-edge is a good option and might not interfere with the growth of the valve. Early results are encouraging but a longer follow-up is needed to evaluate the long-term prognosis. PMID- 19550355 TI - Comparison between carotid-femoral and brachial-ankle pulse wave velocity as measures of arterial stiffness. AB - BACKGROUND: Arterial stiffness is an important risk factor for cardiovascular disease. Carotid-femoral pulse wave velocity (cfPWV) is the most recognized and established index of arterial stiffness. An emerging automatic measure of PWV primarily used in the Asian countries is brachial-ankle PWV (baPWV). METHOD: To systematically compare these two methodologies, we conducted a multicenter study involving a total of 2287 patients. RESULTS: There was a significant positive relation between baPWV and cfPWV (r = 0.73). Average baPWV was approximately 20% higher than cfPWV. Both cfPWV and baPWV were significantly and positively associated with age (r = 0.56 and 0.64), systolic blood pressure (r = 0.49 and 0.61), and the Framingham risk score (r = 0.48 and 0.63). The areas under the receiver operating curves (ROCs) of PWV to predict the presence of both stroke and coronary artery disease were comparable between cfPWV and baPWV. CONCLUSION: Collectively, these results indicate that cfPWV and baPWV are indices of arterial stiffness that exhibit similar extent of associations with cardiovascular disease risk factors and clinical events. PMID- 19550357 TI - Primary subacute Salmonella osteomyelitis of the navicular bone in a child with normal immunity. AB - Primary subacute haematogenous osteomyelitis is one of the causes of limp. It usually involves tubular bones. Flat and small bones are affected less commonly. Diagnosis is difficult and usually takes weeks together for completion. Salmonella spp. can be isolated as a cause of primary subacute haematogenous osteomyelitis, if a usually underlying disorder, such as sickle cell anemia is associated. In this study, we present a child with normal immunity diagnosed as Salmonella primary subacute haematogenous osteomyelitis of the navicular bone, which is a rare condition. Primary subacute haematogenous osteomyelitis must be considered as a cause of limp for timely diagnosis and treatment. PMID- 19550356 TI - Superior consistency of ambulatory blood pressure monitoring in children: implications for clinical trials. AB - BACKGROUND: Casual office blood pressure (CBP) measurements are still standard in antihypertensive drug trials. In pediatric hypertensive trials, ethical considerations, very low disease prevalence and the marked impact of white-coat hypertension create the need for very sensitive and reproducible techniques of BP assessment. We hypothesized that ambulatory BP monitoring (ABPM) may identify treatment effects more sensitively than CBP and thereby reduce sample sizes required in pediatric antihypertensive trials. METHODS: Standard deviations (SDs) were used to assess population variability of CBP and ABPM at baseline and after 6 months standardized antihypertensive treatment from a trial investigating the BP-lowering effect of ramipril in children with chronic kidney disease. RESULTS: In 157 hypertensive children, ramipril had a similar mean BP-lowering effect on clinic and ambulatory 24-h BP for systolic (-10 vs. -11 mmHg, P = NS) and diastolic values (-9 vs. -11 mmHg, P = NS). However, the SDs of the CBP responses were up to 39% larger than those of ABPM (SBP 15.5 vs. 9.4; DBP 13.8 vs. 8.8; both P < 0.0001). Using power analysis, we demonstrate that, depending on the magnitude of the expected antihypertensive effect and trial design, the utilization of ABPM in antihypertensive drug efficacy studies allows reduction of sample sizes by 57-75%. This reduction of cohort size with ABPM is substantially greater than previously observed for adults. CONCLUSION: The primary use of ABPM can substantially reduce the number of children put at potential risk in blinded antihypertensive drug trials by up to three quarters. PMID- 19550358 TI - Torticollis manifest after a minor fall with underlying bony anomalies and a hypoplastic vertebral artery. AB - The etiologies of torticollis are numerous. We describe a unique patient who presented with torticollis with head tilting to the right after sustaining a minor fall. Computed tomography scan with two and three-dimensional reconstructions revealed an underlying hypoplasia of the right lateral mass of C1 and occipital condyle. Further study with MRI/magnetic resonance angiography showed hypoplasia of the right vertebral artery with compensatory dilatation of the left vertebral artery. We postulate these underlying anomalies predisposed the patient to torticollis, which became apparent after his fall. We discuss the surgical options available, and the rationale for a single C1-C2 transarticular screw with iliac crest bone graft and cable. In the literature, there are four reported cases of vascular anomalies resulting in torticollis. Our patient is the first, to our knowledge, who harbored both bony and vascular anomalies. PMID- 19550359 TI - Simultaneous tissue factor expression and phosphatidylserine exposure account for the highly procoagulant pattern of melanoma cell lines. AB - A correlation between cancer and hypercoagulability has been described for more than a century. Patients with cancer are at increased risk for thrombotic complications, and the clotting initiator protein, tissue factor (TF), is possibly involved in this process. In addition to TF, the presence of negatively charged phospholipids, particularly phosphatidylserine (PS), is necessary to support some of the blood-clotting reactions. There are few reports describing PS exposure by tumor cells. In this study, we characterized the procoagulant properties of the murine B16F10 and the human WM-266-4 melanoma cell lines. Flow cytometry analyses showed constitutive TF expression by both cell lines, in contrast to negative staining observed for the nontumorigenic melanocyte lineage, melan-A. In addition, tumor cells accelerate plasma clotting in a number dependent manner. For WM-266-4, this ability was partially reversed by an anti-TF antibody but not by aprotinin, a nonspecific serine-protease inhibitor. Furthermore, flow-cytometric analyses showed the presence of PS at the outer leaflet of both cell lines. This phenomenon was determinant for the assembly of the intrinsic tenase (FIXa/FVIIIa) and prothrombinase (FXa/FVa) complexes, resulting in the activation of FX to FXa and prothrombin to thrombin, respectively. As a result, incubation of WM-266-4 with human plasma produces robust thrombin generation. In conclusion, simultaneous TF expression and PS exposure are responsible for the highly procoagulant pattern of the aggressive melanoma cell lines B16F10 and WM-266-4. Therefore, these cell lines might be regarded as useful models for studying the role of blood coagulation proteins in tumor biology. PMID- 19550360 TI - Nodular malignant melanoma and multiple cutaneous neoplasms under immunosuppression with azathioprine. AB - Immunosuppressed patients are at increased risk of skin cancer. A 67-year-old renal transplant recipient developed a nodular malignant melanoma after 30 years of immunosuppression with azathioprine and prednisolone. The patient died of metastatic disease 3 months after the diagnosis was made. The function of the renal graft was not affected at all. Renal transplant recipients are at high risk of developing nonmelanocytic skin tumors when on immunosuppressive therapy with cyclosporine A. Less common is the development of skin cancer during immunosuppression with azathioprine. Latest reports show the increased incidence of malignant melanoma in immunosuppressed patients. Our case illustrates the necessity of close dermatological surveillance of allograft recipients, to assure an early recognition of any malignant skin tumor and to reduce the risk of systemic metastatic disease. PMID- 19550361 TI - Ca2+ imaging of identifiable neurons labeled by electroporation in insect brains. AB - Identifiable neurons in the silkmoth brain were studied physiologically after loading of Ca indicator by local electroporation. Small groups of neurons with projections in confined regions of the antennal lobe were labeled, and Ca imaging showed differences in the dose-response characteristics between projection neurons of the macroglomerular complex that responded to different pheromone components. Compared with projection neurons, local interneurons showed shorter response latencies. Targeted labeling of neurons innervating restricted regions of neuropil by local electroporation is a powerful method that will be generally useful for elucidating details of the functional circuitry in insect brains. PMID- 19550362 TI - Cardiac repolarization and striatal dopamine transporter function are interrelated. AB - OBJECTIVE: In Parkinson's disease, striatal dopamine transporter (DAT) binding and cardiac sympathetic function are disturbed. In addition, heart rate (HR) corrected cardiac repolarisation time (QTc interval), which is partly under autonomic control, is prolonged. Whether there is physiological coupling between striatal DAT binding and QTc time (QTc-DAT relation) is not known. The purpose of this study is to evaluate QTc-DAT relation in healthy young adults. METHODS: Thirty-five participants (18 women, age 26.4+/-1.8 years; mean+/-SD) were studied with iodine-123 labelled 2beta-carbomethoxy-3beta-(4-iodophenyl) nortropane single photon emission tomography. Signal-averaged ECG was recorded at rest from each participant. QTc interval was computed with Bazett's correction and with the approach by Karjalainen, getting QTc and QTk intervals, respectively. RESULTS: Mean striatal DAT binding, as (striatum-cerebellum)/cerebellum, was 2.63+/-0.31. Mean HR, QT, QTc and QTk intervals were 66+/-9 bpm, 340+/-25 ms, 354+/-18 ms and 351+/-16 ms, respectively. HR-QT correlation was -0.63, P value of less than 0.001. HR was not related to striatal DAT binding. QTc-DAT and QTk-DAT relations were significant, r = -0.50, P = 0.004 and r = -0.59, P = 0.0002, respectively. In linear regression model, striatal DAT binding explained 35% of the variance of QTk interval (95% confidence interval: -46.9 to -13.0, P = 0.0002). CONCLUSION: This study suggests significant physiological QTc-DAT relation in young healthy adults. QTc interval measurements might carry diagnostically important information in clinical conditions, which have an effect on both striatal DAT binding and cardiac sympathetic function. PMID- 19550363 TI - Impact of image reconstruction on phase analysis of ECG-gated myocardial perfusion SPECT studies. AB - OBJECTIVES: Phase analysis (SyncTool) has been developed to assess left ventricular (LV) dyssynchrony from gated myocardial perfusion single-photon emission computed tomography (GSPECT) studies. Conventionally, GSPECT data are reconstructed using filtered backprojection (FBP). This study is intended to determine the impact of various iterative reconstruction methods on SyncTool. METHODS: Thirty consecutive patients, acquired using a Philips CardioMD system, were enrolled in this study. The GSPECT data were reconstructed using FBP, maximum likelihood expectation maximization (MLEM), MLEM with three-dimensional resolution recovery (Astonish), MLEM with Vantage attenuation correction (AC), and MLEM with Vantage AC and three-dimensional Monte Carlo-based scatter correction (ACSC), respectively. The reconstructed data were then submitted to SyncTool to measure LV dyssynchrony (phase standard deviation and histogram bandwidth). The paired t-test was used to compare the LV dyssynchrony indices given by MLEM, Astonish, AC, and ACSC, respectively, with those given by the FBP. RESULTS: No statistical significance was observed for any comparison between iterative reconstruction methods and the FBP. CONCLUSION: Reconstruction methods have insignificant impact on the LV dyssynchrony indices, indicating that the standard FBP reconstruction is sufficient for accurate phase analysis, supporting the widespread clinical use of SyncTool in measuring LV dyssynchrony. PMID- 19550364 TI - Longer-term psychiatric adjustment of children and parents after meningococcal disease. AB - OBJECTIVE: To ascertain whether increases in psychological symptoms in children and parents after meningococcal disease are sustained over time, and to examine the psychosocial and illness associations of 12-mo psychological outcome. DESIGN: A prospective, cohort study using repeated measures. SETTING: Three pediatric intensive care units and 19 general pediatric wards across greater London. PATIENTS: Fifty-six children, aged 3 to 16 yrs, admitted to hospital with meningococcal disease and their parents. MEASURES AND MAIN RESULTS: Child and parent psychological symptoms were measured, using the Strengths and Difficulties Questionnaire (SDQ) and the General Health Questionnaire (GHQ) at three time points: before/during hospital admission, 3 mos, and 12 mos after discharge. The Impact of Event Scale (IES) was used at the two follow-up points. During the follow-up period, there were statistically significant increases over child pre illness levels in parent-rated emotional, conduct, hyperactivity, and impact SDQ scores; the most significant change at 12-mo follow-up was an increase in impact on daily living scores. At 12 mos, five (11%) of 43 children were at risk for posttraumatic stress disorder. The strongest correlations of 12-mo child psychological symptoms (total SDQ scores)--in addition to premorbid total SDQ score--were illness-related changes in parenting, maternal IES and GHQ scores. At 12 mos, 13 (24%) of 54 mothers and six (15%) of 40 fathers scored at high risk for posttraumatic stress disorder. The strongest correlation of maternal posttraumatic stress disorder symptoms (IES scores) was paternal posttraumatic stress disorder symptoms. CONCLUSIONS: Admission to the hospital with meningococcal disease is followed by an increase in psychological symptoms in children at home, some of which are persistent and impairing, and by continuing posttraumatic stress symptoms in a proportion of children and parents. Psychosocial (pre- and postmorbid) factors predict problems at 12-mo follow-up. PMID- 19550365 TI - Surfactant phospholipids, surfactant proteins, and inflammatory markers during acute lung injury in children. AB - OBJECTIVE: To explore the pathophysiology of acute lung injury in children. DESIGN: Prospective cohort study. SETTING: Regional University Hospital, pediatric intensive care unit. PATIENTS: Children without a preexisting lung injury who developed acute lung injury and were intubated were eligible for the study. Children without lung injury and intubated for minor surgical procedures acted as controls. INTERVENTIONS: Bronchoalveolar lavage fluid and blood were collected on days 1 to 4, weekly, and immediately before extubation during acute lung injury. Molecular species compositions of phosphatidylcholine were determined by electrospray ionization mass spectrometry of lipid extracts of bronchoalveolar lavage fluid supernatants. Surfactant proteins A, B, and D and interleukin-8 were measured in bronchoalveolar lavage fluid and plasma by enzyme linked immunosorbent assay and Western blotting. MEASUREMENTS AND MAIN RESULTS: Eighteen children with acute lung injury were enrolled in the study and compared with eight controls. In children with acute lung injury, there were significant changes in the bronchoalveolar lavage fluid phosphatidylcholine species. Bronchoalveolar lavage fluid dipalmitoyl phosphatidylcholine (PC 16:0/16:0) and palmitoyl-myristoyl phosphatidylcholine (PC 16:0/14:0) significantly deceased during acute lung injury (p < .001 and p < .001, respectively), whereas oleoyl linoleoyl PC (18:1/18:2), palmitoyl-linoleoyl PC (16:0/18:2) and stearoyl linoleoyl PC (18:0/18:2) characteristic of plasma PC were significantly increased (p < .05, p < .02, and p < .05 respectively), as well as palmitoyl-oleoyl PC (16:0/18:1), and stearoyl-arachidonoyl PC (18:0/20:4) which are characteristic of cell membranes (p < .02, and p < .02, respectively). There were no significant changes to bronchoalveolar lavage fluid, surfactant protein A or B levels compared with controls during acute lung injury, whereas bronchoalveolar lavage fluid, surfactant protein D, and interleukin-8 levels significantly increased (p < .05 and p < .02, respectively). In plasma during acute lung injury, there were significant increases in surfactant proteins A, B, and D, and interleukin-8 (p < .001, p < .001, p < .05, and p < .001, respectively). CONCLUSION: Changes to the phosphatidylcholine profile, surfactant proteins, and inflammatory markers of bronchoalveolar lavage fluid and plasma in children with acute lung injury are consistent with an alveolar/blood leakage and inflammatory cell membrane degradation products. These changes are due to alveolar capillary membrane damage and cellular infiltration. PMID- 19550366 TI - Multilocus genotypic association with vascular dementia by multifactor dimensionality reduction and entropy-based estimation. AB - OBJECTIVE: We conducted a simultaneous analysis of candidate genetic loci for their genotypic association with the susceptibility to vascular dementia (VaD) to put forth the best model for predicting genetic susceptibility to VaD. METHODS: Individual-locus effects and their epistatic effects on susceptibility to VaD were simultaneously assessed by multifactor dimensionality reduction and entropy based method. The 23 loci in 12 genes were studied in 207 VaD patients and age matched and sex-matched 207 controls. RESULTS: The multifactor dimensionality reduction analysis revealed that the best single-locus candidate model included angiotensinogen (AGT) Thr235Met with testing accuracy (TA) of 58.31%, the best two-locus candidate model included AGT Thr235Met and transforming growth factor beta1 Pro10Leu with TA of 58.06%, the best three-locus candidate model was not significant (P>0.05), and the best four-locus candidate model included transforming growth factor-beta1 Pro10Leu, AGT Thr235Met, sterol regulatory element binding protein 2 G34995T, and leukemia inhibitory factor T4524G with TA of 57.13% (P<0.05). The best four-locus model was, however, still in question because of the inconsistent best model selection by cross-validation. Synergistic epistatic effect of the best two-locus model was proven by entropy-based estimation. CONCLUSION: The best predictor for genetic susceptibility to VaD was the single-locus model of AGT. The best two-locus model reflecting epistasis would be also employed for predicting its susceptibility. Further studies on the epistasis are to elucidate their underlying mechanisms. PMID- 19550368 TI - Utility of diagnostic and prognostic markers in urothelial carcinoma of the bladder. AB - Urothelial carcinoma (UCC) of the bladder demonstrates diverse morphologic features, often leading to diagnostic challenges in the discrimination between UCC and benign mimickers of neoplasia, and between primary UCC and secondary neoplasms involving the bladder. In situ lesions also provide diagnostic difficulty in some instances, most prominently in the distinction between normal, reactive urothelium and flat urothelial carcinoma in situ. The use of ancillary techniques, including panels of immunohistochemical markers, in distinguishing these entities has aided not only in the diagnosis of UCC, but has also provided insight into the molecular pathogenesis and prognostic value of numerous molecular pathways in UCC. This review focuses on some of the more commonly encountered biomarkers in UCC and their role in addressing key diagnostic and prognostic issues in this disease process. PMID- 19550369 TI - Pathologic features and biologic importance of colorectal serrated polyps. AB - Serrated polyps of the large intestine comprise a heterogeneous group of mucosal lesions that includes nondysplastic polyps, such as hyperplastic polyps and sessile serrated polyps, and polyps that show overt cytologic dysplasia, namely serrated adenomas and mixed hyperplastic/adenomatous polyps. These polyps have received increased recognition over the past 2 decades, as emerging evidence suggests that a subset may be precursors to colorectal carcinomas that lack chromosomal instability. Several investigators have proposed the concept of the "serrated neoplastic pathway" according to which nondysplastic serrated lesions develop progressively severe dysplasia culminating in the development of microsatellite unstable carcinomas that show DNA hypermethylation and BRAF mutations. A subset of hyperplastic polyps and sessile serrated polyps show mutations in the BRAF gene and abnormal DNA methylation, which can, ultimately, affect the promoter regions of key DNA-repair and tumor suppressor genes, such as MLH1 and MGMT, leading to their decreased transcription and microsatellite instability. On the basis of this hypothesis, many authors have proposed that sessile serrated polyps should be treated and surveilled similar to conventional adenomas, although prospective data are lacking. This review describes the clinicopathologic and molecular features of serrated polyps and discusses the current data regarding their biologic significance. PMID- 19550370 TI - Carcinomas of the upper aerodigestive tract with rearrangement of the nuclear protein of the testis (NUT) gene (NUT midline carcinomas). AB - This manuscript reviews carcinomas of the upper aerodigestive tract associated with genetic rearrangements of the nuclear protein of the testis (NUT) gene, also known as NUT midline carcinomas. The literature is reviewed regarding all reported cases of NUT midline carcinomas and the clinicopathologic features are discussed. Our current understanding of the molecular pathogenesis of the disease is also discussed, along with the differential diagnosis for undifferentiated or poorly differentiated malignancy of the upper aerodigestive tract. PMID- 19550371 TI - Seeking beyond rejection: an update on the differential diagnosis and a practical approach to liver allograft biopsy interpretation. AB - Pathologic evaluation of liver allograft biopsies plays an integral role in the management of patients after liver transplantation. This review summarizes the clinical context and classical histology of different types of allograft rejection and also the common entities that enter the differential diagnosis of allograft rejection, and provides practical approaches to liver allograft biopsy interpretation. In addition, some of the new developments in the field of liver transplant pathology are updated. The purpose of this review is to provide guidance for pathologists interpreting liver allograft biopsies, particularly those interested in developing expertise in the field of liver transplant pathology. PMID- 19550372 TI - Calretinin: diagnostic utility in the female genital tract. AB - The utility of calretinin was first described in the diagnosis of mesothelioma. In the female genital tract, calretinin was initially reported in Wolffian remnants and related lesions and later used in the diagnosis of sex cord stromal tumors of the ovary, endometrial stromal tumors with sex cord-like differentiation, and uterine tumors resembling sex cord tumors of the ovary. This review discusses calretinin expression in normal tissues of the female genital tract and highlights its potential utility in the diagnosis of these subsets of neoplasms with emphasis to tumors that may constitute a problem in their differential diagnosis. In particular, the limited utility of calretinin in the diagnosis of sex cord stromal tumors of the ovary, in the differential diagnosis between mesothelioma and serous tumors involving the ovary and/or peritoneum, and in the differential diagnosis between Wolffian lesions and endometrial carcinoma are addressed. PMID- 19550373 TI - Glypican-3: a novel diagnostic marker for hepatocellular carcinoma and more. AB - Glypican-3 (GPC3) is a heparan sulfate proteoglycan that plays an important role in cell growth and differentiation. GPC3 function is tissue dependent. In some tissues, GPC3 acts as a tumor suppressor gene, whereas in others, it acts as an oncofetal protein. Studies have shown that GPC3 is a reliable marker for hepatocellular carcinoma. The sensitivity and specificity exceeds both alpha fetoprotein and hepatocyte-paraffin1. GPC3 immunohistochemistry can aid in the differentiation of testicular germ cell tumors, being expressed in all yolk sac tumors but not in seminomas. GPC3 expression has also been identified in some squamous cell carcinomas of the lung and clear cell carcinomas of the ovary. The role of GPC3 in melanomas is still controversial. This article reviews the current information on the application of GPC3 immunostaining in surgical pathology and cytology. PMID- 19550376 TI - Comparative evaluation of lesion enhancement using 1 M gadobutrol vs. 2 conventional gadolinium chelates, all at a dose of 0.1 mmol/kg, in a rat brain tumor model at 3 T. AB - PURPOSE: The purpose of this study is to compare the differences in contrast enhancement using 0.1 mmol/kg body weight 1 M gadobutrol versus 2 standard gadolinium chelates, both formulated at 0.5 M, (gadopentetate dimeglumine and gadoterate meglumine) in a standardized rat brain glioma model at 3 T. MATERIALS AND METHODS: A total of 19 rats were evaluated, divided into 2 groups. Group 1 (n = 10) was examined using gadobutrol and gadopentetate dimeglumine and group 2 (n = 9) was examined using gadobutrol and gadoterate meglumine. The time between the intraindividual injections was at least 24 hours and contrast agent injections were performed in a randomized order. All agents were applied at a dose of 0.1 mmol/kg body weight.Image acquisition was performed using a T1-weighted 2D TSE technique (repetition time/echo time (TE) 500/16, FA 180 degrees) with an acquisition time of 1:47 minutes:seconds. At a field-of-view of 75 x 75 mm2 and a matrix size of 320 x 320, a voxel size of 0.2 x 0.2 x 2.0 mm3 was achieved. Data acquisition was performed before and at 5 consecutive time points every 2 minutes after contrast agent injection. Signal-to-noise ratios (SNRs) of tumor and normal contralateral brain as well as contrast-to-noise ratio (CNR) measurements were performed using region of interest analysis. RESULTS: The increase in tumor contrast enhancement, ranged between 19.6% and 35.9% for gadobutrol versus gadopentetate dimeglumine (group 1) and between 23.2% and 27.8% for gadobutrol versus gadoterate meglumine (group 2). Overall, CNR was statistically significantly higher for gadobutrol in both groups (P < 0.0001). CNR values for gadobutrol were 25.5 +/- 8.2 in group 1 and 27.1 +/- 8.3 in group 2 with respective CNR values for gadopentetate dimeglumine of 18.6 +/- 5.6 and gadoterate meglumine of 19.2 +/- 5.3. At each acquired time point mean values of tumor SNR were higher for gadobutrol (group 1: SNR(mean) range from 78.7-89.1 vs. 74.3-80.8; group 2: SNR(mean) range from 79.9-88.9 vs. 74.2-80.8). Tumor SNR was statistically significant different at all measured time points in group 2 (P < 0.05). In group 1, the difference of tumor SNR was also statistically significant for the gadobutrol/gadopentetate dimeglumine comparison (P < 0.05) with exception of time point at 9 minutes postcontrast (P = 0.07). CONCLUSION: The results of this study show significantly higher brain tumor SNR and CNR postcontrast for gadobutrol compared with gadopentetate dimeglumine and gadoterate meglumine at 3 T. Injecting the same gadolinium chelate dose on a weight basis, tumor mean SNR gains were superior for gadobutrol at all acquired postcontrast time points. This result with gadobutrol may facilitate better brain tumor detection in the presence of blood-brain barrier disruption. PMID- 19550377 TI - Ultra-fast carotid CT-angiography: low versus standard volume contrast material protocol for a 128-slice CT-system. AB - OBJECTIVES: Noninvasive imaging is increasingly accepted for the evaluation of atherosclerotic disease of the carotid arteries. We sought to evaluate the feasibility of a low-contrast media volume protocol for carotid computed tomography angiography (CTA) using a 128-slice-spiral-computed tomography scanner with a gantry rotation time of 300 milliseconds. METHODS AND MATERIALS: Thirty consecutive patients underwent CTA for the evaluation of the carotid vessels, with a 128-section scanner. Fifteen patients were examined with a standard volume contrast injection protocol (group A): 80 mL of contrast material (CM) were injected at 5 mL/s using the test bolus method to assess individual transit time. Another 15 patients were examined with a low-volume contrast media protocol (group B): 30 mL CM were injected at 4 mL/s using bolus tracking to trigger the CTA acquisition. In both groups, contrast administration was followed by a saline flush. Image quality and segmental vascular enhancement as well as the presence and degree of arterial stenosis were independently evaluated by 2 radiologists. Venous enhancement and streak artifacts at the thoracic inlet because of highly concentrated CM in the subclavian veins were evaluated in both groups. Kappa statistic and Pearson correlation coefficient were used to quantify interobserver variability. Qualitative data were compared using the Wilcoxon signed rank test and student t test was used to investigate differences in segmental vessel attenuation. RESULTS: All studies were of diagnostic quality in both groups. Interobserver agreement was high (kappa = 0.82, group A; kappa = 0.78, group B). Attenuation measurement showed excellent interobserver correlation in both groups (r > 0.9). Mean enhancement values were slightly higher in group A, but without statistical significance when averaged for all segments (P = 0.06). Streak artifacts impaired evaluation of 13 adjacent arterial segments in 8 patients at the level of the thoracic inlet in group A. In group B, only 1 segment was rated insufficient by both radiologists. Venous enhancement was significantly lower in group B (P = 0.04). The low-contrast protocol proved to be the more robust method with constant high arterial enhancement, less streak artifacts at the thoracic inlet, and less venous overlay. CONCLUSION: Using the latest CT technology, optimal depiction of the craniocervical arteries can be archived with a low volume (30 mL) CM protocol. PMID- 19550378 TI - Cervical and cranial computed tomographic angiography with automated bone removal: dual energy computed tomography versus standard computed tomography. AB - OBJECTIVE: In supraaortic vessels, bone subtracted maximum intensity projections make the evaluation of computed tomographic angiography (CTA) datasets easier and faster. Dual energy CT can be used for bone removal without user interaction. The purpose of this study was to compare the results of conventional and dual energy based bone removal. MATERIALS AND METHODS: Dual energy CT angiography of the supraaortic vessels was acquired in 30 patients at 140 and 80 kVp tube potential simultaneously. Thick images of 0.75 mm were reconstructed from both datasets, and an additional weighted average dataset using information from both tubes was calculated. Two readers independently assessed vessel delineation, completeness of bone removal, and vessel preservation, as well as adequacy for diagnostic evaluation after dual energy-based bone removal (DEBR) and conventional bone removal (CoBR). The Student t test and Wilcoxon rank sum test were applied to test differences between data for significance. Cohen's kappa-test was used to calculate the interobserver agreement. RESULTS: Of dual energy datasets DEBR 88.3% were rated as adequate for diagnostic evaluation compared with only 6.7% with CoBR, P < 0.001. Of DEBR (35%) contained all vessels and no bony structures, whereas all diagnostic CoBR still contained residual bone and showed partial vessel truncations. Vessel delineation was rated significantly better with DEBR. DEBR profited from stronger vascular enhancement, whereas it had no significant influence on CoBR. Reading times were 173 +/- 55 seconds with DEBR and 253 +/- 12 seconds with CoBR, P < 0.001, which corresponds to a reduction of 32%. Comparison of dual energy CTA versus single energy CTA showed a dose reduction of 29.0% to 43.7%. CONCLUSIONS: Dual Energy-based bone removal can remove bony structures from supraaortic CTA datasets without time-consuming user interaction. This leads to a significant reduction of reading time, radiation dose and improved vessel delineation. PMID- 19550379 TI - For debate: is gender medicine important in pediatrics? PMID- 19550380 TI - Short stature in children with coeliac disease. AB - Coeliac disease (CD) is a genetically determined gluten-sensitive enteropathy resulting in nutrient malabsorption, with an increasing incidence world-wide. Clinical presentation in early childhood may include classic malabsorption symptoms, whereas older CD children often present extra-intestinal symptoms including short stature and pubertal delay. A gluten-free diet (GFD) generally leads to a rapid catch-up in growth and to normalization of the pituitary function. The pathogenesis of CD-associated short stature is still unclear. Besides the involvement of the growth hormone (GH)/insulin-like growth factor-I axis, a role for ghrelin was recently proposed. Furthermore, some CD patients do not show catch-up growth during GFD, despite reversion to seronegativity for CD markers including antiendomysial and anti-tissue transglutaminase antibodies. These subjects show GH deficiency and could potentially benefit from recombinant human GH therapy. This review deals with the management of short stature and the evaluation of growth axis function in CD children. PMID- 19550381 TI - Endocrine disrupters - with special emphasis on sexual development. AB - The impact of environmental pollutants in increasingly observed alterations in wildlife is obvious. Many laboratories and a number of human studies have revealed that human beings are not resistant to these contaminants. We briefly discuss the findings of animal and human studies regarding the effects of endocrine disrupting chemicals relevant to clinical problems encountered in child care, namely, premature and delayed sexual development and disorders of male genital development. PMID- 19550382 TI - Genetic and environmental contributors to cryptorchidism. AB - Cryptorchidism is a common congenital anomaly that shows familial clustering and increased prevalence in first-degree relatives, suggesting that genetic factors contribute to the etiology. Animal models and some human data suggest that environmental exposures may also contribute to risk. Potential variables during development that may contribute to the occurrence of cryptorchidism in the genetically susceptible individual include maternal hormonal influences as well as other environmental factors that affect hormonal signaling. In a subset of affected males, genetic and/or environmental factors may also contribute to associated infertility and malignancy. This review will discuss the limited state of knowledge regarding the etiology of non-syndromic cryptorchidism. Non syndromic cryptorchidism is a common and complex disorder of un known etiology with geographic and perhaps temporal variability (1,2). Although presumed to be multifactorial in etiology, few specific genetic or environmental factors have been clearly linked to the disease in man. Hormonal pathways that participate in testicular descent and the effects of chemical exposures on this process have been studied in animal models. In man (3-5) and animal strains with cryptorchidism (6,7), there is evidence for multilocus genetic susceptibility. However, despite promising gene candidates, few genomic variants have been linked to non-syndromic cryptorchidism. Humans are exposed to a wide variety of potentially anti-androgenic and/or estrogenic environmental chemicals, but the degree to which these may contribute to the pathogenesis of cryptorchidism is not clearly known. Potential factors that may contribute to the etiology of cryptorchidism are discussed. PMID- 19550384 TI - Testicular descent: a hypothesis and review of current controversies. AB - Descent of the testis into the scrotum occurs by a complex multifactorial process involving the normal development of the testis, the hormonal actions of insulin like growth factor 3, testosterone, a intact hypothalamic pituitary testicular axis, the patent processus vaginalis, gubernacular outgrowth and regression and intraabdominal pressure. The paper reviews the key components of testicular descent, the current hypothesis on how testicular descent occurs and the controversies surrounding this hypothesis. PMID- 19550383 TI - Sex steroidal targets & genetic susceptibility to idiopathic cryptorchidism. AB - Cryptorchidism, the most common congenital abnormality in newborn boys, is a major risk factor for male infertility and testicular malignancy in adulthood. This disorder appears as an isolated form or as part of impaired male sex development or a congenital malformation syndrome. Based mainly on the laboratory studies of the rodent models, sex steroidal signaling pathways have been shown to be involved in testicular descent; however, data on the human genetic susceptibility are less compelling. Mutations in the human genes encoding androgen receptor (AR), estrogen receptor alpha (ERalpha) and beta (ERbeta), and steroidogenic factor-1 (SF-1) have occasionally been identified but do not seem to be a frequent cause of this genital malformation. On the other hand, common polymorphisms in these genes have recently been investigated as possible contributing risk factors for idiopathic isolated (nonsyndromic) cryptorchidism, alone or by influencing susceptibility to other causal factors such as environmental endocrine disruptors. ABBREVIATIONS: Androgen Receptor (AR); DNA Binding Domain (DBD) ;Environmental Estrogen Disruptors (EEDs); Estrogen Receptor Alpha (ERalpha); Estrogen Receptor Beta (ERbeta); Ligand-Binding Domain (LBD); Linkage Disequilibrium (LD); Odds Ratio (OR); Restriction Fragment Length Polymorphism (RFLP); Single Nucleotide Polymorphism (SNP); Steroidogenic Factor-1 (SF-1); Transactivation Domain (TAD); Testicular Dysgenesis Syndrome (TDS); Wild Type (WT). PMID- 19550385 TI - Children born small for gestational age or with very low birth weight: clinical similarities and potential benefits of growth hormone therapy. AB - The dynamics of early postnatal 'extrauterine' growth in very low birth weight (VLBW) premature infants suggest a switch from mechanisms supporting normal growth during pregnancy to mechanisms promoting postnatal growth. This may permanently affect endocrinological and metabolic homeostasis and development in some children. Altered neurological development and regulation of glucose metabolism in some children born with VLBW appears similar to children born small for gestational age (SGA). In addition, when present, postnatal growth failure has many similarities in both VLBW infants and infants born SGA after intrauterine growth retardation. At 2 years of age, it is very difficult to distinguish between these groups. Increased awareness, with ongoing assessment, of these children is required. In particular, two research areas need special focus in short VLBW children: the effects of growth hormone treatment (which shows dose-dependent efficacy on postnatal growth in short SGA children), and insulin sensitivity (which seems altered, as in short SGA children). PMID- 19550386 TI - From paediatric to adult endocrinology care: the challenge of the transition period. AB - Interest in the transition period when young people with chronic illness move from a paediatric service to an adult service has been driven by the increasing numbers of young patients making such a move. This brief article offers suggestions for successful transition programmes based on the recent personal experience of the authors, with a review of the available literature. We discuss how physicians need to adopt a flexible approach to the timing of the transfer, how an education programme is needed before the transition to allow patients to manage their illness independently, and how a co-ordinated approach between the paediatric and adult teams is of the utmost importance. The successful approach to transition will not only benefit the patient but, by linking the paediatric and adult services, may aid research into the long-term modalities for hormonal substitution, including thyroid, steroid and growth hormone deficiencies. PMID- 19550387 TI - Abnormal growth in noonan syndrome: the challenge of optimal therapy. AB - Noonan syndrome (NS) is a phenotypically heterogeneous condition frequently associated with short stature. Genetic investigations have identified mutations in several genes, e.g. PTPN11, KRAS, RAF and SOS1 in patients with the NS phenotype and related disorders such as LEOPARD, Costello and Cardiofacio- cutaneous syndromes. In NS, PTPN11 mutations are present in 29-60% of cases. The degree of short stature does not associate closely with the presence of a mutation; however, some PTPN11-positive patients have decreased growth hormone (GH)-dependent growth factors consistent with mild GH insensitivity. GH therapy induces short-term increases in height velocity over 1-3 years, and is likely to improve adult height. PMID- 19550388 TI - Fibromyalgic syndromes: could growth hormone therapy be beneficial? AB - Fibromyalgia is a chronic, idiopathic condition in which patients experience pain, asthenia and fatigue. The pathogenesis of the condition is unknown, and numerous mechanisms have been postulated, including neural hypersensitivity and autoimmunity. Symptoms of fibromyalgia are broadly similar to those of growth hormone deficiency (GHD), and there is evidence of decreased GH secretion and functional GHD in a subset of patients with fibromyalgia. Use of GH therapy in this patient population therefore represents a rational treatment strategy. Preliminary placebo-controlled trials have shown that GH therapy can significantly improve signs and symptoms of fibromyalgia and quality of life in patients receiving the current standard of care. Despite the use of relatively high doses of GH in these patients, treatment is well tolerated. Several mechanisms of action for GH in fibromyalgia have been suggested, including both central and peripheral effects. PMID- 19550389 TI - Growth hormone deficiency after traumatic brain injury in adults: when to test and how to treat? AB - Hypopituitarism has numerous potential causes, and it is becoming clear that traumatic brain injury (TBI), including traffic accidents and sport-related injuries, is commonly associated with pituitary dysfunction. Mechanisms of pituitary damage after TBI include direct injury and vascular problems, and more recent research suggests that autoimmunity may also be involved. There may also be a genetic influence, as the E3 allele of the ApoE gene may provide some protection from post-traumatic hypopituitarism. Studies suggest that patients with mild or moderate TBI are likely to recover pituitary function over time. In patients with severe TBI, however, adrenocorticotrophic hormone and growth hormone deficiencies may persist. Patients who experience TBI should, therefore, be followed up carefully and evaluated for pituitary dysfunction to ensure that appropriate hormone replacement therapy can be provided if needed. PMID- 19550390 TI - Benefits from growth hormone therapy after bariatric surgery. AB - There is increasing interest in the use of bariatric surgery to achieve weight loss in individuals with severe obesity. However, there are concerns over the loss of lean body mass (LBM) following surgery in individuals who have functional growth hormone (GH) deficiency related to their obesity. We discuss here the results of an open-label study in which we studied the efficacy of GH treatment to prevent loss of LBM and metabolic disturbances in 24 women with functional severe GH deficiency who were undergoing laparoscopic adjustable gastric banding for severe obesity. Participants were randomized to standard care (including a dietary restriction programme) with or without GH treatment, and were studied for 6 months. Overall, weight loss and reductions in waist circumference were similar in the two groups. GH treatment was associated with a significant decrease in fat mass and a significantly smaller decrease in LBM, as well as improvements in insulin sensitivity, and lipid profile. PMID- 19550391 TI - Adherence remains a challenge for patients receiving growth hormone therapy. AB - Patients receiving growth hormone therapy have the best growth results when they have good adherence, but adherence rates are often low. Adherence has been difficult to measure and is often measured indirectly as direct methods are inconvenient, expensive or both. However, use of electronic monitors that log the dose and the time it was delivered provides a simple and reliable means of gathering data and may facilitate the discussion of adherence in the clinic. Although many factors impact on adherence, there are some 'red flags' to which a physician should be particularly alert, such as history of poor attendance at consultations. Based on the author's own clinical experience and opinion, as well as published studies, this article describes factors associated with non adherence and how most, if not all, barriers to good adherence can be overcome by maintaining a good non-judgemental relationship with the patient, and delivering useful and clear education and training soon after diagnosis. PMID- 19550392 TI - Improving adherence to GH therapy with an electronic device: first experience with easypod. AB - Endocrinology units in the UK and Eire have, in recent years, moved away from offering a narrow range of injection devices and products to patients starting growth hormone (GH) treatment, and now most (approximately 90%) offer some form of patient choice. Survey results from a unit in Birmingham, UK, have shown that it is not possible to predict which device will be selected by a given patient based on their sex, age or diagnostic category. Across the UK, there is, however, wide variability in the methods used to present the devices to patients, the time invested in reaching an informed decision and the range of devices and manufacturers offered. Patients not offered a free choice of device at the start of treatment have been shown to be less likely to adhere to treatment, and this is associated with diminished height velocity. These results show the importance of offering patients a choice in their GH delivery device. PMID- 19550394 TI - Nuclear receptors and their role in Epstein -- Barr virus induced B cell transformation. AB - Epstein - Barr virus (EBV) is a lymphotropic virus that infects more than 90% of the human population, and targets B cells for infection. Infection of human B cells leads to the malignant transformation and eventual immortalization. In latency III infection six EBV-encoded nuclear antigens (EBNAs) and three latent membrane proteins (LMPs) are expressed in the transformed cells that can grow as a lymphoblastoid cell lines in vitro . These proteins hijack the normal B cell growth pathways by activating the constitutive growth promotion and external survival signals. We have determined a set of the nuclear receptors that are up- (and down-) regulated in the latency III infected cells at the mRNA level. In the present paper we discussed the possible role of these receptors in B cell transformation upon EBV infection based on the literature data. PMID- 19550395 TI - Expression of galectin-1 in malignant tumors. AB - Galectin-1 is a 14 kDa lectin expressed ubiquitously in mammalian organism. Since its discovery, the lectin was shown to participate in many cellular processes. Involvement in some of them like induction of apoptosis of activated T cells, mediation of cell adhesion and participation in angiogenesis suggest that galectin-1 could be utilized by malignant tumors. Indeed expression of galectin-1 is upregulated in tumors of different origin. Many examples point to its important role in a process of cancer metastasis. This review summarizes the data available to date on galectin-1 expression in human malignancies. PMID- 19550396 TI - Tumor hypoxia and malignant progression. AB - The current problems of malignant growth biology, in particular the molecular background of the specific microenvironment of tumor cells and their interaction with stromal cells, which mediates the behavior of tumors and the tumor-host interrelationship were the subject of the International Conference entitled "Tumor Hypoxia and Malignant Progression", a meeting held at the House of Scientists of the NAS of Ukraine in Kyiv, Ukraine, October 1 st to 4 th , 2008. The meeting was hosted by the R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology of the NAS of Ukraine (IEPOR), and was dedicated to the 90 th Anniversary of the National Academy of Sciences of Ukraine. Over the last years, scientists have focused extensively on the problem of tumor hypoxia as a factor promoting tumor progression. It is known that hypoxia, as a constituent of the tumor cell microenvironment as well as aerobic glycolysis, are important features of malignant tumors. The direct correlation between high levels of hypoxia and tumor aggressiveness has been shown in numerous studies. Therefore, hypoxia is regarded as a factor of unfavorable prognosis. There is a number of different methods available for the evaluation of the level of hypoxia, some of which are being applied in the clinical setting. The stimulating impact of hypoxia and hypoxia-associated proteins on neoangiogenesis and vasculogenesis in tumor tissue has been demonstrated. Several studies have focused on the development of agents capable of blocking hypoxia-associated signaling pathways and vasculogenesis in tumor. Recently, the direct association between hypoxia dependent signaling pathways and expression of factors that mediate inflammation in tumor tissue, in particular tumor-associated macrophages has been shown. To summarize, a better understanding of the relationships between hypoxia-associated signaling pathways, metabolic peculiarities and inflammatory factors that positively influence tumor progression may elucidate not only how the aggressive tumor phenotype is formed but also may assist in the development of new approaches for the treatment of cancer patients. PMID- 19550397 TI - Molecular profile and cell cycle in MCF-7 cells resistant to cisplatin and doxorubicin. AB - AIM: To compare ultrastructure, phenotypic profile and cell cycle progression of MCF-7 human breast cancer cells and MCF7 sublines resistant to cisplatin (MCF 7/DDP) and doxorubicin (MCF-7/DOX). METHODS: MTT-test, immunocytochemistry, flow cytometry, electron microscopy. RESULTS: The development of drug resistance to cisplatin and doxorubicin in MCF-7 cells upon the culturing of the initial cells with the raising concentrations of cytostatics was accompanied by the increase in cells adhesion, the increasing differentiation grade and the loss of steroid hormone receptors. Besides, it was shown that antiapoptotic mechanisms (decrease of Bcl-2 expression) and intracellular glutathione detoxifying system are involved in the process of cisplatin resistance development in MCF-7 cells. At the same time, P-glycoprotein overexpression in cells resistant to doxorubicin suggests MDR-dependent mechanism. Both doxorubicin- and cisplatin-resistant cells are characterized by the changes in the expression of several cell cycle regulators -- Ki-67, cyclin D1, pRb and p21). CONCLUSION: The long-time culture of MCF-7 cells with cytostatic drugs results in the decreased cyclin D1, pRb, and Ki-67 expression and increased p21 expression with the increasing differentiation grade of the resistant cells. The underlying mechanisms of resistance to cisplatin and doxorubicin in MCF-7 cells may be different. PMID- 19550398 TI - Expression profile of nuclear receptors upon Epstein -- Barr virus induced B cell transformation. AB - BACKGROUND: Infection of human B cells with Epstein - Barr virus (EBV) induces metabolic activation, morphological transformation, cell proliferation and eventual immortalization. AIM: To identify the nuclear receptors, which are the cellular interaction partners of EBNAs, that will help to elucidate the mechanism of B cell transformation. METHODS: We have compared the nuclear receptor profile in the naive and EBV-transformed B-lymphocytes, using TaqMan LDA microfluidic card technology. RESULTS: Out of 48 nuclear receptor, 17 showed differential expression at the mRNA level. The expression of 5 genes was elevated in EBV transformed cells, whereas 12 genes were downregulated in lymphoblastoid cells (LCLs). 7 genes were studied at the protein level; 2 genes were up regulated (Nr2F2 and RARA) and 4 genes were down regulated (ERB, NUR77, PPARG, and VDR) in LCLs. CONCLUSION: The nuclear receptor profiling on EBV infected B cells showed alterations of nuclear receptors expression at both mRNA and protein levels compared with non infected peripheral blood cells. Further analysis on a possible role of each nuclear receptor in EBV induced cell transformation should be performed. PMID- 19550399 TI - Blood serum immunoglobulins of patients with multiple myeloma are capable of hydrolysing histone H1. AB - BACKGROUND: Recently we have shown that the imunoglobulins G from blood serum of some multiple sclerosis patients are capable of cleaving histone H1. AIM: To check whether histone H1-hydrolyzing abzymes could be detected not only in blood plasma of autoimmune patients, but also during cancer development, particularly during the onset of multiple myeloma. METHODS: Immunoglobulines were isolated from blood serum of multiple myeloma patients (n = 11) by precipitation with 50% ammonium sulfate and tested for proteolytic activity toward linker and core calf thymus histones. Antibody preparations able to cleaved histone H1 were subjected to affinity chromatography on histone H1-Sepharose with following analysis of chromatographic fractions' protease activity. To prove that antibody molecules are responsible for hydrolysis of histone H1, gel filtration at acidic pH with subsequent examination of protease activity of chromatographic fractions (pH shock analysis) was used. RESULTS: It was found that 3 of 11 antibody preparations are capable of hydrolyzing calf thymus histone H1 but not core histones. It was shown that histone H1-hydrolysing activity of 2 proteolytically active antibody preparations is associated with IgGs that possess affinity towards histone H1. pH-shock analysis proved that protease activity towards histone H1 is intrinsic property of IgG molecules. CONCLUSIONS: We demonstrated the existence of previously unknown histone H1 hydrolyzing IgG abzymes in the serum of multiple myeloma patients. Possible biological role of hisH1-hydrolyzing antibodies in patients with multiple myeloma was discussed. PMID- 19550400 TI - Expression of cathepsin L in nasopharyngeal carcinoma and its clinical significance. AB - AIM: To study the expression of cathepsin L in nasopharyngeal carcinoma (NPC), and analyze its relationship with clinicopathologic factors. METHODS: The expression of cathepsin L was determined immunohistochemically in NPC, adjacent normal nasopharyngeal tissues and metastatic cervical lymph nodes. The correlation between its protein expression and clinicopathologic parameters as well as with long term follow-up data in NPC was analyzed. RESULTS: The protein of cathepsin L was overexpressed in 47% primary tumor tissues, and in 89% metastatic cervical lymph node samples. Overexpression of cathepsin L was found to correlate with lymph node metastasis ( P = 0.04) and distant metastasis ( P = 0.01), and marginally with clinical stage and T classification, but not with patient age, gender and histological classification of tumor. Patients with overexpression of cathepsin L in tumor tissue had worse 5-year survival than those without such expression pattern ( P = 0.033). Multivariate survival analysis showed that cathepsin L protein expression level had a marginal significant correlation with prognosis of NPC. CONCLUSION: Cathepsin L is a potential biomarker for prognosis of NPC and contributes to NPC metastasis. PMID- 19550401 TI - Image fusion using CT, MRI and PET for treatment planning, navigation and follow up in percutaneous RFA. AB - AIM: To evaluate the feasibility of fusion of morphologic and functional imaging modalities to facilitate treatment planning, probe placement, probe re positioning, and early detection of residual disease following radiofrequency ablation (RFA) of cancer. METHODS: Multi-modality datasets were separately acquired that included functional (FDG-PET and DCE-MRI) and standard morphologic studies (CT and MRI). Different combinations of imaging modalities were registered and fused prior to, during, and following percutaneous image-guided tumor ablation with radiofrequency. Different algorithms and visualization tools were evaluated for both intra-modality and inter-modality image registration using the software MIPAV (Medical Image Processing, Analysis and Visualization). Semi-automated and automated registration algorithms were used on a standard PC workstation: 1) landmark-based least-squares rigid registration, 2) landmark based thin-plate spline elastic registration, and 3) automatic voxel-similarity, affine registration. RESULTS: Intra- and inter-modality image fusion were successfully performed prior to, during and after RFA procedures. Fusion of morphologic and functional images provided a useful view of the spatial relationship of lesion structure and functional significance. Fused axial images and segmented three-dimensional surface models were used for treatment planning and post-RFA evaluation, to assess potential for optimizing needle placement during procedures. CONCLUSION: Fusion of morphologic and functional images is feasible before, during and after radiofrequency ablation of tumors in abdominal organs. For routine use, the semi-automated registration algorithms may be most practical. Image fusion may facilitate interventional procedures like RFA and should be further evaluated. PMID- 19550402 TI - Assessment of anthracycline-induced cardiotoxicity with electrocardiography. AB - AIM: Monitoring of anthracycline-induced cardiotoxicity with electrocardiography (ECG) and comparing ECG changes with findings on echocardiography (ECHO). METHODS: A total of 26 adult acute leukemia patients (mean age 46.2 -/+ 12.4 years, 15 males) treated with 2-6 cycles of anthracycline-based chemotherapy (CT) were studied. Cardiac evaluation was performed at the baseline (before CT), after first CT, after last CT (cumulative anthracycline dose 464.3 -/+ 117.5 mg/m2) and circa 6 months after CT. Time ECG parameters, QRS voltage, presence of repolarization changes, arrhythmias and other abnormalities were evaluated. RESULTS: During treatment and follow-up, we found a statistical significant QTc interval prolongation - 414.7 -/+ 16.0 ms (before CT), 419.6 -/+ 21.6 ms (after first CT), 428.0 -/+ 16.2 ms (after last CT) and 430.1 -/+ 18.4 ms (6 months after CT). Significant QTc interval prolongation (> 450 ms) occurred in 3 patients after first CT, in 4 patients after last CT and in 5 patients within 6 months after CT. Significant total QRS voltage lowering in the limb leads (> 1.0 mV versus before CT) occurred in 3 patients after first CT, in 5 patients after last CT and in 6 patients within 6 months after CT. We found a statistically significant correlation between decreased QRS voltage, QTc interval prolongation and left ventricular (LV) dysfunction on ECHO. Repolarization changes associated with oncology treatment were present in 9 patients within 6 months after CT. CONCLUSION: Anthracycline treatment is associated with changes in electrical activity of the myocardium. Prolonged QTc interval represents a risk for development of malignant ventricular arrhythmias. Decreased QRS voltage and prolonged QTc interval after anthracycline treatment could correlate with LV dysfunction on ECHO. Further studies will be needed to prove whether these ECG changes could serve as an accessible and non-invasive screening method indicating LV dysfunction after anthracycline treatment. PMID- 19550403 TI - Analysis of in vitro and in vivo sensitivity of oral cancer cells to methotrexate. AB - AIM: The present study was directed on the assessment of the response of treatment-naive oral cancer cells to methotrexate (MTX) in vitro: and clinical response to MTX therapy. METHODS: A pilot study of in vitro: evaluation of MTX response of oral cancer cells from 10 patients was conducted using a cell viability assay to determine the sensitivity/resistance to MTX. Quantitative in vitro: data were correlated to the clinical outcome to MTX therapy. RESULTS: A positive correlation was observed between the effect of MTX on tumor cells in vitro: and clinical response for 7 out of 10 patients. CONCLUSIONS: Observations from the proof-of-principle pilot study suggests that oral cancer cells have intrinsically variable response to MTX. Confirmation of these findings with a larger cohort of patients could aid in the development of individualized therapies for this class of malignancy. PMID- 19550404 TI - Cardiovascular changes associated with infusion of hematopoietic cell grafts in oncohematological patients -- impact of cryopreservation with dimethylsulfoxide. AB - AIM: Dimethylsulfoxide (DMSO) is the most frequently used agent for hematopoietic cell (HC) graft cryopreservation. This study aimed to monitor blood pressure and heart rate (HR) during HC graft infusion and assess the impact of cryopreservation with DMSO. METHODS: 153 HC graft infusions in 153 consecutive hematological patients (mean age 49.1 -/+ 12.6 years; 80 males) were evaluated. Cryopreservation with DMSO was used in 133 grafts (DMSO group). Twenty grafts were infused directly without cryopreservation (control group). Systolic blood pressure (SBP), diastolic blood pressure (DBP) and HR were measured immediately before and after HC graft infusion. RESULTS: SBP and DBP increased significantly after graft infusions cryopreserved with DMSO ( p<0.0001 for SBP; p<0.01 for DBP). Increases (> 10mmHg) in SBP were seen in 42 (31.6%) patients; in DBP in 31 (23.3%) patients. Changes in HR were non-significant in DMSO group. Increases in BP and HR correlated with increasing DMSO dose (p<0.01; p<0.05, respectively). Changes in SBP, DBP and HR were non-significant in control group. CONCLUSION: HC graft infusions cryopreserved with DMSO could cause statistically significant increases in SBP and DBP, without changes in HR. These changes were mostly transient and asymptomatic, not requiring therapeutic intervention. However, they might cause complications, especially in patients with preexisting cardiovascular disease, who should be monitored closely during HC transplantation. PMID- 19550406 TI - Demonstration of cutaneous allodynia in association with chronic pelvic pain. AB - Pelvic pain is a common condition that is associated with dysmenorrhea and endometriosis. In some women the severe episodes of cyclic pain change and the resultant pain becomes continuous and this condition becomes known as Chronic Pelvic Pain. This state can be present even after the appropriate medical or surgical therapy has been instituted. It can be associated with pain and tenderness in the muscles of the abdomen wall and intra-pelvic muscles leading to severe dyspareunia. Additional symptoms of irritable bowel and interstitial cystitis are common. A common sign of the development of this state is the emergence of cutaneous allodynia which emerges from the so-called viscero-somatic reflex. A simple bedside test for the presence of cutaneous allodynia is presented that does not require excessive time or special equipment. This test builds on previous work associated with changes in sensation related to gall bladder function and the viscera-somatic reflex(1;2). The test is undertaken with the subject s permission after an explanation of how the test will be performed. Allodynia refers to a condition in which a stimulus that is not normally painful is interpreted by the subject as painful. In this instance the light touch associated with a cotton-tipped applicator would not be expected to be painful. A positive test is however noted by the woman as suddenly painful or suddenly sharp. The patterns of this sensation are usually in a discrete pattern of a dermatome of the nerves that innervate the pelvis. The underlying pathology is now interpreted as evidence of neuroplasticity as a consequence of severe and repeating pain with changes in the functions of the dorsal horns of the spinal cord that results in altered function of visceral tissues and resultant somatic symptoms(3). The importance of recognizing the condition lies in an awareness that this process may present coincidentally with the initiating condition or after it has been treated. It also permits the clinician to evaluate the situation from the perspective that alternative explanations for the pain may be present that may not require additional surgery. PMID- 19550405 TI - Expression of drug resistance proteins in triple-receptor-negative tumors as the basis of individualized therapy of the breast cancer patients. AB - AIM: To evaluate the efficacy of the application of various chemotherapy schemes based on the immunohistochemical study of expression patterns of proteins associated with the drug resistance P-glycoprotein (P-gp), glutathione-S transferase (GST), metallothioneins (MT) of breast cancer (BC) patients with the triple-receptor-negative (RE(-), RP(-), HER-2/neu(-)) cancer. METHODS AND RESULTS: P-gp, GST and MT expression in BC-biopsy samples from 60 BC patients was evaluated by immunohistochemical analysis. The results of the clinical observations showed that 3-years relapse-free survival rate of the patients of with P-gp, GST and MT-positive tumors treated with taxoter + adriablastin / taxoter + cyclophosphamide (TA/TC), gemcitabine + carboplatin, or TC + bevacizumab was 61.5%, 78.6% and 81.2% respectively, vs 41.2% in the control group with P-gp, GST and MT-negative tumors treated with adriablastin + cyclophosphamide (p<0.05), while overrall suvival rates were 84.4%, 92.6% and 93.8% respectively vs 70.6% in the control group (p<0.05). CONCLUSION: The study points on the possibility to elevate the efficiency of polychemotherapy by its individualization based on the expression patterns of P-gp, GST and MT on tumor cells of the patients with the triple-receptor-negative BC. PMID- 19550407 TI - Oral rehydration solutions in non-cholera diarrhea: a review. AB - The use of oral rehydration solution (ORS) has revolutionized the management of acute diarrhea. The implementation of the standard World Health Organization ORS (WHO-ORS) has resulted in decreased mortality associated with acute diarrheal illnesses in children, although in general stool volume and diarrhea durations are not reduced. Decreased morbidity and mortality have occurred because of improved hydration status. Decreased morbidity has also been described in adults who used this therapy. Various modifications to the standard ORS have been derived. These modifications have included hypo-osmolar or hyperosmolar solutions, use of rice-based ORS, zinc supplementation, and the use of amino acids, including glycine, alanine, and glutamine. Some of these variations have been successful, some have not, and others are still under investigation. ORS has been used for travelers' diarrhea and to decrease intravenous (IV) fluid requirements in patients with short bowel syndrome (SBS) who require parenteral nutrition (PN). This paper reviews the standard WHO-ORS and its mechanism of action, followed by more contemporary reduced osmolarity ORS and rice-based ORS in non-cholera diarrhea. Various modifications to improve ORS are also discussed. PMID- 19550408 TI - Light and moderate alcohol consumption significantly reduces the prevalence of fatty liver in the Japanese male population. AB - OBJECTIVES: The effect of alcohol consumption on the liver is controversial. Recent reports have suggested that moderate alcohol consumption decreases the prevalence of elevated alanine aminotransferase levels. The role of alcohol consumption in the development of fatty liver (FL), however, has not been studied definitively. The aim of this study was to examine the association between alcohol consumption and FL in a large Japanese population. METHODS: A total of 7,431 asymptomatic male subjects who underwent a complete medical survey in our institute between May 2007 and July 2008 were recruited. Cases positive for hepatitis B or C viruses, potential hepatotoxic drug intake, or under treatment for metabolic disorders were excluded. FL was defined by ultrasonography. Visceral and subcutaneous adipose tissues (VAT and SAT) were measured by computed tomography. Independent and significant predictors associated with FL were determined by multiple logistic regression analysis. RESULTS: Of the initial study candidates, 130 (1.7%) were positive for hepatitis B and 66 (0.8%) were positive for hepatitis C. On the basis of the inclusion and exclusion criteria, 5,599 men (50.9+/-8.1 years) were studied cross-sectionally. Light (40-140 g/week) and moderate (140-280 g/week) alcohol consumption significantly and independently reduced the likelihood of FL (odds ratio=0.824 and 0.754, 95% confidence interval=0.683-0.994 and 0.612-0.928, P=0.044 and 0.008, respectively) by multivariate analysis after adjusting for potential confounding variables. VAT, SAT, low-density lipoprotein, triglycerides, and fasting blood glucose were significant predictors of the increased prevalence of FL, whereas age was a predictor of the decreased prevalence of FL. CONCLUSIONS: The prevalence of FL was significantly and independently decreased by light and moderate alcohol consumption in men of an asymptomatic Japanese population. PMID- 19550409 TI - A population-based study of breastfeeding in inflammatory bowel disease: initiation, duration, and effect on disease in the postpartum period. AB - OBJECTIVES: We aimed to assess breastfeeding practices and the impact of breastfeeding on disease flare during the postpartum year in inflammatory bowel disease (IBD). METHODS: Women of childbearing age from 1985 to 2005 were identified from the University of Manitoba IBD Research Registry. Questionnaires were completed regarding pregnancy and the postpartum period. Data for initiation and duration of breastfeeding were compared with population-based regional data. RESULTS: Of 204 eligible women, 132 (64.7%) responded to the survey, yielding information on 156 births. Breastfeeding was initiated in 83.3% of women with IBD (n=132), 81.9% of Crohn's disease patients (CD, n=90), and 84.2% of ulcerative colitis patients (UC, n=39) vs. 77.1 % in the general population (P>0.05 for all). Of women with IBD, 56.1% breastfed for >24 weeks vs. 44.4% of controls (P=0.02). The rate of disease flare in the postpartum year was 26% for those who breastfed vs. 29.4% in those who did not (P=0.76) in CD and 29.2% vs. 44.4% (P=0.44) in UC. The odds ratio of disease flare postpartum for those who breastfed vs. those who did not was 0.58 (95% CI: 0.24-1.43), 0.84 (0.19-9.87), and 0.51 (0.12-2.2) for IBD total, CD, and UC, respectively. Risk of disease flare was not related to age at pregnancy, duration of disease, or socioeconomic status. CONCLUSIONS: Women with IBD are as likely as the general population to breastfeed their infants. Breastfeeding is not associated with an increased risk of disease flare and may even provide a protective effect against disease flare in the postpartum year. PMID- 19550410 TI - Risk factors for abnormal liver function tests in patients with ileal pouch-anal anastomosis for underlying inflammatory bowel disease. AB - OBJECTIVES: Liver involvement is common in patients with inflammatory bowel disease (IBD). However, the frequency and the significance of liver function test (LFT) abnormalities in patients with ileal pouch-anal anastomosis (IPAA) for underlying IBD have not been studied. The aim of this study was to evaluate the prevalence and to identify risk factors for abnormal LFTs in patients with IPAA and underlying IBD. METHODS: All patients were identified from our prospectively maintained Pouchitis Database between 2002 and 2008. Abnormal LFTs were classified as the following: (i) any abnormal elevation of transaminases, and/or alkaline phosphatase (ALP), and/or bilirubin; (ii) hepatitis, if there was more than twice the elevation of transaminases; and (iii) cholestatic, if there was more than 1.5 times elevation of ALP. Clinical, endoscopic, and histological variables were assessed using Cox proportional hazard models for evaluating risk for abnormal LFTs. RESULTS: A total of 545 IPAA patients with underlying IBD were identified from the database, of which 373 patients who had LFTs done after their pouch surgery were included. This included 346 patients with ulcerative colitis, 25 with indeterminate colitis, and 2 with Crohn's colitis before surgery. Their mean age was 45.9+/-13.8 years. A total of 65 patients (17.4%) (40 men, 25 women, median age: 47 years) had abnormal LFTs. Of the patients, 52 (13.9%) had abnormal transaminases, whereas 15 (4%) were classified as having hepatitis. Thirty-five (9.4%) patients had an abnormal ALP level, with 18 (4.8%) classified as cholestatic. The most common cause of an abnormal LFT was transient elevation in 32 (49.2%) patients, followed by fatty liver (fatty change on imaging with body mass index (BMI) > or =25 kg/m(2) in the absence of other causes, including alcohol abuse and drug-induced hepatitis) in 10 (15.4%), drug-induced abnormal LFTs in 7 (10.7%), and chronic hepatitis B or C in 6 (9.2%). Primary sclerosing cholangitis (PSC) was responsible for abnormal LFTs in 10 patients (15.4%). Cox proportional hazard model analysis showed that BMI (hazard ratio (HR)=1.07, 95% confidence interval (95% CI): 1.02, 1.12; P=0.003), the presence of PSC (HR=4.49, 95% CI: 1.45, 13.89; P=0.009), autoimmune disorder (HR=2.54, 95% CI: 1.09, 5.93; P=0.031), a family history of IBD (HR=2.32, 95% CI: 1.29, 4.17; P=0.005), and extensive colitis before colectomy (HR=4.59, 95% CI: 2.04, 10.33; P<0.001) predicted any abnormal LFTs. CONCLUSIONS: Abnormal LFTs were common in patients with IPAA in this cohort. The presence of co-existing autoimmune disorder, a family history of IBD, extensive colitis before colectomy, the presence of PSC, and a high BMI appear to be a significant risk factors for abnormal LFTs. Whether abnormal LFTs affect health-related quality of life, pouch survival, and selection of pouch-related medical therapy requires further investigation. PMID- 19550411 TI - Alcohol intake, alcohol dehydrogenase genotypes, and liver damage and disease in the Danish general population. AB - OBJECTIVES: We tested the hypothesis that alcohol, alone and in combination with alcohol dehydrogenase (ADH) 1B and ADH1C genotypes, affects liver damage and disease in the general population. METHODS: Information on alcohol intake and on liver disease was obtained from 9,080 men and women from the Copenhagen City Heart Study. Biochemical tests for the detection of liver damage were specific for alanine aminotransferase (ALT), aspartate aminotransferase (AST)-to-ALT ratio (AST/ALT), gamma-glutamyl transpeptidase (gamma-GT), albumin, bilirubin, alkaline phosphatase, coagulation factors, and erythrocyte volume. RESULTS: Increasing alcohol intake was associated with increasing erythrocyte volume, AST/ALT, and levels of ALT, gamma-GT, albumin, bilirubin, coagulation factors, and with decreasing levels of alkaline phosphatase. Multifactorially adjusted hazard ratios for alcoholic liver disease overall were 0.9 (95% confidence interval (CI), 0.6-1.4), 1.4 (0.8-2.5), 1.8 (0.9-3.5), and 4.1 (2.5-7.0) for an alcohol intake of 1-13, 14-20, 21-27, and > or = 28 drinks per week, respectively, compared with drinking <1 drink per week (P for trend<0.0001); the corresponding hazard ratios for alcoholic liver cirrhosis were 1.7 (0.6-4.7), 2.0 (0.8-7.1), 6.5 (2.0-21), and 13 (4.6-37) (P for trend<0.0001). ADH1B and ADH1C genotypes were not associated with and did not modify the effect of alcohol on biochemical tests or risk of liver disease. CONCLUSIONS: Increasing alcohol intake from none to low (1-6 drinks per week) through to moderate (7-20 drinks per week) and excessive intake (> or = 21 drinks per week) leads to stepwise increases in signs of liver damage with no threshold effect, and to an increased risk of liver disease. The minor changes in biochemical tests for low alcohol intake may not account for subclinical liver disease. PMID- 19550412 TI - Mortality rate and risk factors in patients with hereditary pancreatitis: uni- and multidimensional analyses. AB - OBJECTIVES: Patients with hereditary pancreatitis (HP) bear a high risk of pancreatic adenocarcinoma, but their life expectancy remains unknown. The objective of the study was to assess whether the high risk of cancer decreases survival. METHODS: Inclusion criteria were the presence of a PRSS1 mutation with pancreatic symptoms or chronic pancreatitis in at least two first-degree relatives or three second-degree relatives without another cause. Survival rates were assessed according to risk factors. Excess mortality compared with the general French population was calculated (statistical Esteve model) for two periods (20-50 and 50-70 years), according to several risk factors. RESULTS: The cohort comprised 189 patients. PRSS1 mutations were found in 66%. A total of 19 patients died at the median age of 60. In all, 10 deaths were attributable to HP, including 8 to pancreatic adenocarcinoma. Median overall survival for the whole cohort was 74 years (95% confidence interval (CI): 71-79). The presence of R122H mutation, gender, tobacco consumption in patients older than 18 years, and diabetes mellitus were not associated with differences in survival. Only patients with pancreatic cancer had decreased survival (P=0.008). Excess mortality risk compared with the general population was 0.02% between 20 and 50 years, and 0.61% between 50 and 70 years (NS). Gender, R122H mutation, diabetes, and tobacco use were not associated with excess mortality in these two periods. CONCLUSIONS: Despite their high risk of cancer, HP patients do not have excess mortality risk compared with the general population, irrespective of gender, tobacco use, or diabetes mellitus. These data should be brought to the patient's attention. PMID- 19550413 TI - An analysis of the factors associated with the development of complications in patients undergoing precut sphincterotomy: a prospective, controlled, randomized, multicenter study. AB - OBJECTIVES: Precut is performed when biliary access at endoscopic retrograde cholangiopancreatography (ERCP) fails. Precut may have adjunctive risks, but some authors have suggested that the attempts to cannulate the papilla that precede precutting cause complications. We evaluated the role of the timing of precut in determining the development of complications and with respect to the other factors involved. METHODS: During ERCP, after 10 min of attempts to cannulate, patients were randomized to an early-precut group (n=77) undergoing precut immediately or a late-access group (n=74) in which cannulation was attempted for 10 further minutes before the endoscopist was free to perform precut or to persist in cannulation. Occurrence of complications and the associated risk factors were recorded. RESULTS: The two groups were similar for general characteristics. The number of attempts to cannulate, the number of pancreas injections, and the incidence of acinarization were higher in the late-access group. The cannulation rate was 94%. The incidence of overall complications was similar, but the pancreatitis rate was higher in the late-access group (14.9 vs. 2.6%, P=0.008). Amylase levels increased by 398.9+/-879.4 in the early-precut group and 833.6+/-1478.4 in the late-access group (P=0.029). Nondilated bile duct and pancreatic injection were related to the development of pancreatitis, whereas the performance of precut was related to other complications. CONCLUSIONS: Early precut is associated with lower pancreatitis rate, suggesting that pancreatitis develops as a consequence of the attempts to cannulate the papilla and pancreatic injection, and not precutting. PMID- 19550414 TI - Effect of systemic alcohol and nicotine on airway protective reflexes. AB - OBJECTIVES: Injection of water into the pharynx induces contraction of the upper esophageal sphincter (UES), triggers the pharyngo-UES contractile reflex (PUCR), and at a higher volume, triggers an irrepressible swallow, the reflexive pharyngeal swallow (RPS). These aerodigestive reflexes have been proposed to reduce the risks of aspiration. Alcohol ingestion can predispose to aspiration and previous studies have shown that cigarette smoking can adversely affect these reflexes. It is not known whether this is a local effect of smoking on the pharynx or a systemic effect of nicotine. The aim of this study was to elucidate the effect of systemic alcohol and nicotine on PUCR and RPS. METHODS: Ten healthy non-smoking subjects (8 men, 2 women; mean age: 32+/-3 s.d. years) and 10 healthy chronic smokers (7 men, 3 women; 34+/-8 years) with no history of alcohol abuse were studied. Using previously described techniques, the above reflexes were elicited by rapid and slow water injections into the pharynx, before and after an intravenous injection of 5% alcohol (breath alcohol level of 0.1%), before and after smoking, and before and after a nicotine patch was applied. Blood nicotine levels were measured. RESULTS: During rapid and slow water injections, alcohol significantly increased the threshold volume (ml) to trigger PUCR and RPS (rapid: PUCR: baseline 0.2+/-0.05, alcohol 0.4+/-0.09; P=0.022; RPS: baseline 0.5+/-0.17, alcohol 0.8+/-0.19; P=0.01, slow: PUCR: baseline 0.2+/-0.03, alcohol 0.4+/-0.08; P=0.012; RPS: baseline 3.0+/-0.3, alcohol 4.6+/-0.5; P=0.028). During rapid water injections, acute smoking increased the threshold volume to trigger PUCR and RPS (PUCR: baseline 0.4+/-0.06, smoking 0.67+/-0.09; P=0.03; RPS: baseline 0.7+/ 0.03, smoking 1.1+/-0.1; P=0.001). No similar increases were noted after a nicotine patch was applied. CONCLUSIONS: Acute systemic alcohol exposure inhibits the elicitation PUCR and RPS. Unlike cigarette smoking, systemic nicotine does not alter the elicitation of these reflexes. PMID- 19550415 TI - Efficacy of adalimumab in moderate-to-severe pediatric Crohn's disease. AB - OBJECTIVES: The use of tumor necrosis factor-alpha (TNF-alpha) antagonists has changed the therapeutic strategy for Crohn's disease (CD). Adalimumab (ADA), a fully human anti-TNF-alpha monoclonal antibody, is an effective therapy for patients with CD, both naive patients and those intolerant or refractory to Infliximab (IFX), a chimeric anti-TNF-alpha agent. However, the use of ADA is rarely reported in pediatric CD. We performed an open prospective evaluation of short- and long-term efficacy and safety of ADA in children with moderate-to severe CD. METHODS: A total of 23 pediatric CD patients (9 naive and 14 intolerant or unresponsive to IFX) received ADA subcutaneously as a loading schedule at weeks 0 and 2, and at every other week (eow) during a 48-week maintenance phase. Loading and maintenance doses were 160/80 and 80 mg eow in 13 cases, 120/80 and 80 mg eow in 2, and 80/40 and 40 mg eow in 8 cases. The primary efficacy outcomes were clinical remission and response at different scheduled visits along the maintenance phase. At baseline, 13 patients also received immunomodulators (IMs). RESULTS: At weeks 2, 4, 12, 24, and 48, remission rates were 36.3, 60.8, 30.5, 50, and 65.2%, respectively, whereas response rates were 87, 88, 70, 86, and 91%, respectively. Four patients at week 24 and 2 at week 48 received IMs; the mean daily corticosteroid dose, disease activity index, C reactive protein level, and erythrocyte sedimentation rate decreased significantly throughout the trial. No serious adverse events were recorded. CONCLUSIONS: ADA can be an effective and safe biological agent for inducing and maintaining remission in children with moderate-to-severe CD, even in those with previous IFX therapy. PMID- 19550416 TI - AJG series: molecular biology for clinicians. AB - Pharmacogenetics is an evolving field that provides the link between an individual's genetic code and drug metabolism and drug response. This field offers the great promise of individualized medication selection and optimized dosage to maximize treatment response and to minimize adverse side effects. As our understanding of the role of the effects of genetic variants on drug metabolism and body drug processing grows, so does our ability to educate and inform our patients about expected treatment response to the medications being prescribed to them. This brief review will provide an overview of genetics, pharmacogenetics, and current and future examples of genetic variants predicting drug response in gastrointestinal disease, and the limitations and the promise of this exciting and developing field. PMID- 19550417 TI - Fatty acids, IL6, and TNFalpha polymorphisms: an example of nutrigenetics in Crohn's disease. AB - OBJECTIVES: The aim of this work was to study the interaction between genetic polymorphisms (single-nucleotide polymorphisms, SNPs) of pro- and anti inflammatory cytokines and fat intake on the risk of developing Crohn's disease (CD) or modifying disease activity. METHODS: Seven SNPs in interleukin 1 (IL1), tumor necrosis factor alpha (TNFalpha), lymphotoxin alpha (LTalpha), and IL6 genes were analyzed in 116 controls and 99 patients with CD. The type of fat intake was evaluated, and the interaction between SNPs and dietary fat in modulating disease activity was analyzed. RESULTS: Individuals who were homozygous for the IL6-174G/C polymorphism had a six-fold higher risk for CD (odds ratio (OR)=6.1; 95% confidence interval (95% CI)=1.9-19.4), whereas the TT genotype on the TNFalpha-857C/T polymorphism was associated with more active disease (OR=10.4; 95% CI=1.1-94.1). A high intake of total, saturated, and monounsaturated fats, as well as a higher ratio of n-6/n-3 polyunsaturated fatty acid (PUFA), was associated with a more active phenotype (P<0.05). Furthermore, there was an interaction between dietary fat intake and SNPs, with a high intake of saturated and monounsaturated fats being associated with active disease, mainly in patients carrying the variant alleles of the 857 TNFalpha polymorphism (OR=6.0, 95% CI=1.4-26.2; OR=5.17; 95% CI=1.4-19.2, respectively) and the 174 IL6 polymorphism (OR=2.95; 95% CI=1.0-9.1; OR=3.21; 95% CI=1.0-10.4, respectively). Finally, low intake of n-3 PUFA and high n-6/n-3 PUFA ratio in patients with the TNFalpha 857 polymorphism were associated with higher disease activity (OR=3.6; 95% CI=1.0-13.0; OR=5.92; 95% CI=1.3-26.5, respectively). CONCLUSIONS: These results show that different types of fat may interact with cytokine genotype, modulating disease activity. PMID- 19550418 TI - Missed opportunities to initiate endoscopic evaluation for colorectal cancer diagnosis. AB - OBJECTIVES: Delayed diagnosis of colorectal cancer (CRC) is among the most common reasons for ambulatory diagnostic malpractice claims in the United States. Our objective was to describe missed opportunities to diagnose CRC before endoscopic referral, in terms of patient characteristics, nature of clinical clues, and types of diagnostic-process breakdowns involved. METHODS: We conducted a retrospective cohort study of consecutive, newly diagnosed cases of CRC between February 1999 and June 2007 at a tertiary health-care system in Texas. Two reviewers independently evaluated the electronic record of each patient using a standardized pretested data collection instrument. Missed opportunities were defined as care episodes in which endoscopic evaluation was not initiated despite the presence of one or more clues that warrant a diagnostic workup for CRC. Predictors of missed opportunities were evaluated in logistic regression. The types of breakdowns involved in the diagnostic process were also determined and described. RESULTS: Of the 513 patients with CRC who met the inclusion criteria, both reviewers agreed on the presence of at least one missed opportunity in 161 patients. Among these patients there was a mean of 4.2 missed opportunities and 5.3 clues. The most common clues were suspected or confirmed iron deficiency anemia, positive fecal occult blood test, and hematochezia. The odds of a missed opportunity were increased in patients older than 75 years (odds ratio (OR)=2.3; 95% confidence interval (CI) 1.3-4.1) or with iron deficiency anemia (OR=2.2; 95% CI 1.3-3.6), whereas the odds of a missed opportunity were lower in patients with abnormal flexible sigmoidoscopy (OR=0.06; 95% CI 0.01-0.51), or imaging suspicious for CRC (OR=0.3; 95% CI 0.1-0.9). Anemia was the clue associated with the longest time to endoscopic referral (median=393 days). Most process breakdowns occurred in the provider-patient clinical encounter and in the follow up of patients or abnormal diagnostic test results. CONCLUSIONS: Missed opportunities to initiate workup for CRC are common despite the presence of many clues suggestive of CRC diagnosis. Future interventions are needed to reduce the process breakdowns identified. PMID- 19550419 TI - A population-based cohort study on sun habits and endometrial cancer. AB - BACKGROUND: No large cohort study has examined the risk of endometrial cancer in relation to sun exposure. METHODS: A population-based cohort study of 29,508 women who answered a questionnaire in 1990-92, of whom 24,098 responded to a follow-up enquiry in 2000-02. They were followed for an average of 15.5 years. RESULTS: Among the 17,822 postmenopausal women included, 166 cases of endometrial cancer were diagnosed. We used a multivariate Cox regression analysis adjusting for age and other selected demographic variables to determine the risk of endometrial cancer. Women using sun beds >3 times per year reduced their hazard risk (HR) by 40% (0.6, 95% confidence interval (CI) 0.4-0.9) or by 50% when adjusting for body mass index or physical activity (HR 0.5, 95% CI 0.3-0.9), and those women who were sunbathing during summer reduced their risk by 20% (HR 0.8 95% CI 0.5-1.5) compared with women who did not expose themselves to the sun or to artificial sun (i.e., sun beds). CONCLUSION: Exposure to artificial sun by the use of sun beds >3 times per year was associated with a 40% reduction in the risk of endometrial cancer, probably by improving the vitamin D levels during winter. PMID- 19550420 TI - Inhibition of Akt sensitises neuroblastoma cells to gold(III) porphyrin 1a, a novel antitumour drug induced apoptosis and growth inhibition. AB - BACKGROUND: Gold(III) porphyrin 1a is a new class of anticancer drug, which inhibits cell proliferation of wide range of human cancer cell lines and induces apoptosis in human nasopharyngeal carcinoma cells. However, the underlying signalling mechanism by which gold(III) porphyrin 1a modifies the intracellular apoptosis pathways in tumour cells has not been explained in detail in neuroblastoma cells. METHODS: Cell proliferation and apoptosis were determined by measuring 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Annexin V binding, respectively. Western blot assay was used to detect proteins involved in apoptotic and Akt pathways. In vivo tumour growth was assessed by inoculating tumour cells to nude mice subcutaneously, and gold(III) porphyrin 1a was administrated intravenously. RESULTS: This study assessed the antitumour effect and mechanism of gold(III) porphyrin 1a on neuroblastoma in vitro and in vivo. Gold(III) porphyrin 1a displayed a growth inhibition and induction of apoptosis in neuroblastoma cells effectively in vitro, which was accompanied with release of cytochrome c and Smac/DIABLO and caspases activation. Further studies indicated that gold(III) porphyrin 1a inhibited X-linked inhibitor of apoptosis (XIAP). However, we found that gold(III) porphyrin 1a can induce a survival signal, Akt activation within minutes and could last for at least 24 h. To further confirm association between activation of Akt and the effectiveness of gold(III) porphyrin 1a, neuroblastoma cells were treated with API-2, an Akt specific inhibitor. API-2 sensitised cells to gold(III) porphyrin 1a-induced apoptosis and growth inhibition. CONCLUSION: These results suggested that Akt may be considered as a molecular 'brake' that neuroblastoma cells rely on to slow down gold(III) porphyrin 1a-induced apoptosis and antiproliferation. Gold(III) porphyrin 1a is a mitochondrial apoptotic stimulus but also activates Akt, suggesting an involvement of Akt in mediating the effectiveness to growth inhibition and apoptosis by gold(III) porphyrin 1a and that inhibition of Akt can enhance the anticancer activity of gold(III) porphyrin 1a in neuroblastoma. PMID- 19550421 TI - Epstein-Barr virus antibody level and gastric cancer risk in Korea: a nested case control study. AB - BACKGROUND: Few cohort studies have investigated Epstein-Barr virus (EBV) infection before the occurrence of gastric cancer. METHODS: Among 14,440 cohort participants, 100 incident gastric cancer cases were individually matched to two controls. Epstein-Barr virus antibodies IgG and IgA against viral capsid antigen (VCA), EBV nuclear antigen (EBNA) antibody IgG, and early antigen (EA) antibody IgG were measured using enzyme immunoassays (EIAs). RESULTS: The highest titres of VCA IgG (odds ratio (OR): 1.37, 95% confidence interval (CI): 0.62-3.06) or EBNA IgG (OR: 0.87, 95% CI: 0.51-1.46) were not associated with gastric cancer risk. CONCLUSION: Higher levels of VCA IgG or EBNA IgG were not associated with increased risk of gastric adenocarcinoma in Koreans. PMID- 19550422 TI - Identification of macrophage migration inhibitory factor and human neutrophil peptides 1-3 as potential biomarkers for gastric cancer. AB - BACKGROUND: Proteomic methods have the potential to meet the urgent need for better cancer biomarkers. We have used a range of proteomic analyses of serum and tissue from gastric cancer patients and relevant controls to discover biomarkers for gastric cancer. METHODS: Surface-enhanced laser desorption/ionisation time-of flight mass spectrometry (SELDI) and antibody arrays were used to compare protein expression in 21 pairs of gastric cancer tissue and adjacent normal mucosa and serum from 51 gastric cancer patients and 29 patients with benign gastric diseases. Expression differences were confirmed by enzyme-linked immunosorbent assay. RESULTS: Tissue analysis shows human neutrophil peptides 1-3 (HNPs 1-3) elevated 10-fold (P=0.001) in gastric cancer relative to adjacent normal mucosa. Macrophage migration inhibitory factor (MIF) was increased five-fold (P=1.84 x 10(-7)) in the serum of gastric cancer patients relative to individuals with benign gastric disease. The large increase in MIF concentration in serum gives an area under the receiver operating characteristic curve of 0.85. CONCLUSIONS: Proteomic analyses of serum and tissue indicate that HNPs 1-3 and MIF have potential as biomarkers for gastric cancer. In particular MIF may be useful, either alone or in combination with other markers, for diagnosing and monitoring gastric cancer. PMID- 19550423 TI - Factors affecting attitudes toward colorectal cancer screening in the primary care population. AB - BACKGROUND: Colorectal cancer (CRC) is a major cause of death in the United Kingdom. Regular screening could significantly reduce CRC-related morbidity and mortality. However, screening programmes in the United Kingdom have to date seen uptake rates of less than 60%. Attitudes towards screening are the primary factors determining patient uptake. METHODS: A questionnaire was sent to people aged 50-69 years who were registered with general practices in the West Midlands. A total of 11 355 people (53%) completed the questionnaire. Multivariable logistic regression analyses were performed to identify those factors (gender, age, ethnicity, deprivation, number of symptoms, and their duration) that most strongly contributed to negative/positive attitudes in the primary care population. RESULTS: Fourteen percent of respondents had a negative attitude towards screening. Men, older people, and those with Indian ethnic backgrounds were more likely to have negative attitudes toward screening, whereas people with Black-Caribbean ethnic background, people with multiple symptoms and those reporting abdominal pain, bleeding, and tiredness were more likely to have a positive attitude. CONCLUSION: Culturally relevant screening strategies should aim to increase knowledge of the symptoms and signs related to bowel cancer among South Asian ethnic groups in the United Kingdom. It is also important to find ways to increase the acceptability of screening among asymptomatic patients. PMID- 19550424 TI - Chemoprevention of lung carcinogenesis in addicted smokers and ex-smokers. AB - Chemoprevention of lung carcinogenesis is one approach to controlling the epidemic of lung cancer caused by cigarette smoking. The target for chemoprevention should be the activities of the multiple carcinogens, toxicants, co-carcinogens, tumour promoters and inflammatory compounds in cigarette smoke. At present there are many agents, both synthetic and naturally occurring, that prevent lung tumour development in well-established animal models. It seems likely that logically constructed mixtures of these agents, developed from the ground up, will be necessary for the prevention of lung carcinogenesis. PMID- 19550425 TI - Apoptosis and cancer: the genesis of a research field. AB - In multicellular organisms, the total number of cells is a balance between the cell-generating effects of mitosis and cell death that is induced through apoptosis. A disruption of this delicate balance can lead to the development of cancer. This Timeline article focuses on how the field of apoptosis biology has developed in the context of its contribution to our understanding of cell death, or lack of it, in the development of malignant disease. It traces the course of research from key discoveries in fundamental biology to potential therapeutic applications. PMID- 19550427 TI - The reliability of a food frequency questionnaire for use among adolescents. AB - BACKGROUND: Accurate measurement of dietary intake is essential for understanding the long-term effects of adolescent diet on chronic disease risk. However, adolescents may have limited food knowledge and ability to quantify portion sizes and recall dietary intake. Therefore, food frequency questionnaires (FFQs) deemed appropriate for use among adults may not be suitable for adolescents. OBJECTIVES: To evaluate an FFQ in comparison with a 3-day food record (FR) in 14-year olds participating in a population-based cohort study in Western Australia. METHODS: Nutrient intakes estimated by a semi-quantitative FFQ were compared with those from a 3-day FR using Bland & Altman limits of agreement (LOA), tertile classifications and Pearson's correlation coefficients. RESULTS: A total of 785 adolescents provided data from both dietary methods. Mean agreement between the FR and FFQ ranged from 73 (starch) to 161% (vitamin C). The LOA ranged from 27 (retinol) to 976% (carotene), with most nutrients being overestimated by the FFQ. For most nutrients, agreement between the two methods varied significantly with the magnitude of intake. Pearson's r ranged from 0.11 (polyunsaturated fats) to 0.52 (riboflavin). The FFQ classified 80 to 90% of subjects' nutrient intakes into the same or adjacent tertile as their FR. Boys performed slightly better for all of these indices. CONCLUSIONS: Agreement between individual FFQ and FR nutrient intakes was less than ideal. However, the FFQ was able to correctly rank a reasonable proportion of adolescents. PMID- 19550426 TI - Wildlife cancer: a conservation perspective. AB - Until recently, cancer in wildlife was not considered to be a conservation concern. However, with the identification of Tasmanian devil facial tumour disease, sea turtle fibropapillomatosis and sea lion genital carcinoma, it has become apparent that neoplasia can be highly prevalent and have considerable effects on some species. It is also clear that anthropogenic activities contribute to the development of neoplasia in wildlife species, such as beluga whales and bottom-dwelling fish, making them sensitive sentinels of disturbed environments. PMID- 19550428 TI - Dose-dependent effects of decaffeinated coffee on endothelial function in healthy subjects. AB - BACKGROUND/OBJECTIVES: Coffee is known to contain antioxidant substances whose effects may be blunted because of caffeine that may unfavorably affect the cardiovascular system. This study was designed to investigate the acute dose dependent effects of decaffeinated coffee (DC) on endothelial function measured by the brachial artery flow-mediated dilation (FMD). SUBJECTS/METHODS: A total of 15 (8 men and 7 women) healthy nonobese subjects underwent a single-blind, crossover study. Subjects ingested one and two cups of decaffeinated Italian espresso coffee in random order at 5- to 7-day intervals. RESULTS: In the hour following the ingestion of two cups of DC, FMD increased (mean+/-s.e.m.): 0 min, 7.4+/-0.7%; 30 min, 8.0+/-0.6%; 60 min, 10.8+/-0.8%; P<0.001) as compared to consumption of one cup of DC (0 min, 6.9+/-0.7%; 30 min, 8.4+/-1.2%; 60 min, 8.5+/-1.1%; 3 x 2 repeated-measures analysis of variance: P=0.037 for time x treatment effect). Blood pressure did not differ between groups, and basal heart rate was lower in the two-cup group at baseline and 60 min. CONCLUSIONS: The present study demonstrated a significant acute favorable dose-dependent effect of decaffeinated espresso coffee on endothelial function. Further studies are needed to investigate the effects of chronic use of DC especially with respect to caffeinated coffee and in subjects with cardiovascular diseases. PMID- 19550429 TI - Folate intake and the risk of colorectal cancer in a Korean population. AB - BACKGROUND: Folate, a water-soluble B vitamin and one of the major micronutrients in vegetables, is known as an essential factor for the de novo biosynthesis of purines and thymidylate, and it plays an important role in DNA synthesis and replication. Thus, folate deficiency results in ineffective DNA synthesis, and has been shown to induce the initiation and progression of colorectal cancer (CRC). Recently, the incidence of CRC in Korea has increased markedly in both men and women; this trend may be related to the adoption of a more 'westernized' lifestyle, including dietary habits. OBJECTIVE: A hospital-based case-control study was conducted to examine the relationship between folate intake and the risk of CRC within a Korean population. METHODS: A total of 596 cases and 509 controls, aged 30-79 years, were recruited from two university hospitals. Site- and sex-specific odds ratios (ORs) were estimated using logistic regression models. RESULTS: Cases were more frequently found to have a family history of CRC among first-degree relatives, to consume more alcohol, to be more likely current smokers and less likely to participate in vigorous physical activity than the controls. In the overall data for men and women combined, multivariate ORs (95% confidence interval (CI), P for trend) comparing the highest vs the lowest quartile of dietary folate intake were: 0.47 (0.32-0.69, <0.001) for CRC, 0.42 (0.26-0.69, <0.001) for colon cancer and 0.48 (0.28-0.81, 0.007) for rectal cancer. An inverse association was also found in women with dietary folate intake: 0.36 (0.20-0.64, <0.001) for CRC, 0.34 (0.16-0.70, 0.001) for colon cancer and 0.30 (0.12-0.74, 0.026) for rectal cancer, but not in men. In addition, the total folate intake of women was strongly associated with a reduced risk of rectal cancer (OR, 0.38; 95% CI, 0.17-0.88; P for trend=0.04). CONCLUSION: We found a statistically significant relationship between higher dietary folate intake and reduced risk of CRC, colon cancer and rectal cancer in women. A significant association is indicated between higher total folate intake and reduced risk of rectal cancer in women. PMID- 19550430 TI - Beer consumption and the 'beer belly': scientific basis or common belief? AB - BACKGROUND/OBJECTIVES: The term 'beer belly' expresses the common belief that beer consumption is a major determinant of waist circumference (WC). We studied the gender-specific associations between beer consumption and WC (partially in relation to body weight and hip circumference (HC) change). PARTICIPANTS/METHODS: Within the European Prospective Investigation into Cancer and Nutrition (EPIC) Potsdam study (7876 men, 12 749 women), cross-sectional associations were investigated applying general linear models. Prospective analyses of baseline beer consumption and an 8.5-year WC change were assessed using multivariate general linear models and polytomous logistic regression. To test the site specific effect of beer consumption on WC, an adjustment for concurrent changes in body weight and HC was carried out. In addition, the relationship between change in beer consumption and change in WC was studied. RESULTS: A positive association in men and no association in women were seen between beer consumption and WC at baseline. Men consuming 1000 ml/d beer were at 17% higher risk for WC gain compared with very light consumers. Significantly lower odds for WC gain (odds ratio=0.88; 95% confidence interval 0.81, 0.96) were found in beer abstaining women than in very-light-drinking women. The adjustment for concurrent body weight and HC change diminished effect estimates notably, explaining most of the association between beer and change in WC. Decreasing beer consumption was related to higher relative odds for WC loss, although not statistically significant. CONCLUSIONS: Beer consumption leads to WC gain, which is closely related to concurrent overall weight gain. This study does not support the common belief of a site-specific effect of beer on the abdomen, the beer belly. PMID- 19550431 TI - Walking and body mass index in a Portuguese sample of adults: a multilevel analysis. AB - Physical inactivity is an important risk factor for many chronic diseases. The purpose of this study was to investigate the cross-sectional associations between walking and body mass index (BMI). This study comprised 9991 adults (5723 women), aged 37.8+/-9.5 years, from the 2004 Azorean Physical Activity and Health Study. Walking was assessed with the International Physical Activity Questionnaire, and expressed as minutes per week. BMI was calculated from self-reported weight and height. A series of multilevel linear regression models were fitted to assess regression coefficients and s.e. predicting BMI. Results show that, in both genders, and after adjustments for potential confounders, walking was not a significant predictor of BMI. Therefore, our analysis does not extend the findings of earlier studies as it shows no significant associations between walking and BMI, after adjustments for potential confounders. Nevertheless, among Azoreans walking should be encouraged, as walking has other health benefits, beyond controlling obesity. PMID- 19550432 TI - Pregnancy-related changes in activity energy expenditure and resting metabolic rate in Switzerland. AB - BACKGROUND/OBJECTIVES: To measure resting metabolic rate (RMR), activity energy expenditure (AEE), total energy expenditure (TEE) and physical activity pattern, that is, duration and intensity (in metabolic equivalents, METs) of activities performed in late pregnancy compared with postpartum in healthy, well-nourished women living in Switzerland. SUBJECTS/METHODS: Weight, height, RMR, AEE, TEE and physical activity patterns were measured longitudinally in 27 healthy women aged 23-40 years at 38.2+/-1.5 weeks of gestation and 40.0+/-7.2 weeks postpartum. RESULTS: The RMR during late pregnancy was 7480 kJ per day, that is, 1320+/-760 kJ per day (21.4%) higher than the postpartum RMR (P<0.001). Absolute changes in RMR were positively correlated with the corresponding changes in body weight (r=0.61, P<0.001). RMR per kg body weight was similar in late pregnancy vs postpartum (P=0.28). AEE per kg during pregnancy and postpartum was 40+/-13 and 50+/-20 kJ/kg, respectively (P=0.001). There were significant differences in daily time spent at METs<1.5 (1067 vs 998 min, P=0.045), at 2.5< or =METs <3.0 (58 vs 82 min, P=0.002) and METs> or =6 (1 vs 6 min, P=0.014) during pregnancy and postpartum, respectively. CONCLUSIONS: Energy expenditure in healthy women living in Switzerland increases in pregnancy compared with the postpartum state. Additional energy expenditure is primarily attributed to an increase in RMR, which is partly compensated by a decrease in AEE. The decrease in physical activity-related energy costs is achieved by selecting less demanding activities and should be taken into account when defining extra energy requirements for late pregnancy in Switzerland. PMID- 19550433 TI - Dietary fibre intake is inversely associated with carotid intima-media thickness: a cross-sectional assessment in the PREDIMED study. AB - OBJECTIVE: To assess the association between the intake of dietary fibre and carotid intima-media thickness (IMT) in a Mediterranean population at high cardiovascular risk. METHODS: Baseline cross-sectional assessment of 457 men and women (average age 67 years) from two different Spanish centres of the PREDIMED trial. A previously validated food frequency questionnaire (137 food items) was administered by trained dieticians in a face-to-face interview. Mean common carotid IMT was measured using B-mode ultrasound imaging of the right and left carotid arteries by four certified sonographers who used a common protocol. Anthropometric and blood pressure measurements were performed and samples of fasting blood were obtained. Participants were categorized into four groups (roughly quartiles: < or =21; >21 to < or =25; >25 to < or =31 and >31 g/day) of energy-adjusted intake of dietary fibre. Multiple linear regression models were used to adjust for age, sex, centre, smoking, body mass index, diabetes, blood pressure, lipid levels and statin use. RESULTS: In the crude analyses, energy adjusted fibre intake showed a significant inverse correlation with IMT (r=-0.27, P<0.001). In multivariate analyses, a modest, though statistically significant (P=0.03) inverse association between energy-adjusted fibre intake and IMT was also found. The multivariate-adjusted difference in average IMT was -0.051 mm (95% confidence interval: -0.094 to-0.009, P=0.02) for participants whose intake was >35 g/day, (n=47) when compared with those whose intake was <25 g/day (n=224). CONCLUSIONS: Our results suggest that high fibre intake is inversely associated with carotid atherosclerosis. PMID- 19550434 TI - Diet and glycosylated haemoglobin in the 1946 British birth cohort. AB - OBJECTIVES: Raised glycosylated haemoglobin (HbA(1c)) concentration is a recognized risk factor for diabetes, the incidence of which is rising worldwide. The intake of certain foods has been related to HbA(1c) concentration. The aim of this study was to investigate whether nutrient intake, sourced by these foods, was predictive of raised glycosylated haemoglobin (HbA(1c)) concentration in a British cohort. SUBJECTS: The subjects were 495 men and 570 women who were members of the Medical Research Council National Survey of Health and Development, 1946 birth cohort.Diet was assessed from 5-day records in 1982, 1989 and 1999. HbA(1c) was measured in blood samples collected in 1999. Individuals in whom concentration of HbA(1c) was > or =6.3% were identified as being 'at risk' and their nutrient intake was compared with those whose concentration of HbA(1c) was within the normal range (< or =6.2%). RESULTS: Lower intakes of protein, carbohydrate, non-starch polysaccharide, iron, folate, vitamin B(12) and a higher percentage energy from fat in 1989 were significantly predictive of high HbA(1c) status in 1999. In 1999, there were no nutrient intakes that were predictive of HbA(1c) status. Global tests of whether the intakes of energy, carbohydrate, sodium, iron, riboflavin and vitamin B(12) at all three time points were related to HbA(1c) status in 1999, were significant. CONCLUSION: An increased intake of energy, carbohydrate, sodium, iron, riboflavin and vitamin B(12) over 10 years was predictive of raised HbA(1c) status. Increased energy intake may have resulted in increase in body weight, which is a risk factor for diabetes. PMID- 19550435 TI - Beckwith-Wiedemann syndrome. AB - Beckwith-Wiedemann syndrome (BWS) is a model disorder for the study of imprinting, growth dysregulation, and tumorigenesis. Unique observations in this disorder point to an important embryonic developmental window relevant to the observations of increased monozygotic twinning and an increased rate of epigenetic errors after subfertility/assisted reproduction. PMID- 19550436 TI - Comparing population structure as inferred from genealogical versus genetic information. AB - Algorithms for inferring population structure from genetic data (ie, population assignment methods) have shown to effectively recognize genetic clusters in human populations. However, their performance in identifying groups of genealogically related individuals, especially in scanty-differentiated populations, has not been tested empirically thus far. For this study, we had access to both genealogical and genetic data from two closely related, isolated villages in southern Italy. We found that nearly all living individuals were included in a single pedigree, with multiple inbreeding loops. Despite F(st) between villages being a low 0.008, genetic clustering analysis identified two clusters roughly corresponding to the two villages. Average kinship between individuals (estimated from genealogies) increased at increasing values of group membership (estimated from the genetic data), showing that the observed genetic clusters represent individuals who are more closely related to each other than to random members of the population. Further, average kinship within clusters and F(st) between clusters increases with increasingly stringent membership threshold requirements. We conclude that a limited number of genetic markers is sufficient to detect structuring, and that the results of genetic analyses faithfully mirror the structuring inferred from detailed analyses of population genealogies, even when F(st) values are low, as in the case of the two villages. We then estimate the impact of observed levels of population structure on association studies using simulated data. PMID- 19550438 TI - Public health interpretation of trihalomethane blood levels in the United States: NHANES 1999-2004. AB - Trihalomethanes (THMs) can form as byproducts during drinking water disinfection, which is crucial for limiting human exposure to disease-causing pathogens. The US Environmental Protection Agency (USEPA), recognizing both the importance of water disinfection for public health protection and potential risks associated with THM exposure, developed disinfection byproduct rules with the parallel goals of ensuring safe drinking water and limiting the levels of THMs in public water systems. The National Health and Nutrition Examination Survey (NHANES) THM blood data can be used as a means for assessing US population exposures to THMs; biomonitoring equivalents (BEs) can provide human health risk-based context to those data. In this paper, we examine the blood THM levels in the 1999-2004 NHANES data to (i) determine weighted population percentiles of blood THMs, (ii) explore whether gender and/or age are associated with blood THM levels, (iii) determine whether temporal trends can be discerned over the 6-year timeframe, and (iv) draw comparisons between population THM blood levels and BEs. A statistically significant decrease in blood chloroform levels was observed across the 1999-2004 time period. Age-related differences in blood chloroform levels were not consistent and no gender-related differences in blood chloroform levels were observed. The concentrations of all four THMs in the blood of US residents from the 2003 to 2004 NHANES dataset are below BEs consistent with the current US EPA reference doses. For bromodichloromethane and dibromochloromethane, the measured median blood concentrations in the United States are within the BEs for the 10(-6) and 10(-4) cancer risk range, whereas measured values for bromoform generally fall below the 10(-6) cancer risk range. These assessments indicate that general population blood concentrations of THMs are in a range considered to be a low to medium priority for risk assessment follow-up, according to the guidelines for interpretation of biomonitoring data using BEs. PMID- 19550439 TI - Urine-sampling methods for environmental chemicals in infants and young children. AB - This review paper examines and evaluates urine-sampling methodologies in infants and young children, to determine which methods are suitable for use in large biomonitoring surveys or studies of environmental chemicals in children younger than 6 years. Methods for non-toilet-trained children include the use of urine bags, collection pads (e.g., cotton or gauze inserts), disposable diapers, cotton diapers, and the clean catch method. In toilet-trained children, collection methods include use of a commode insert pan as well as specimen collection cups. The advantages and disadvantages of these various methods need to be evaluated with respect to the target population, timing and frequency of collection, minimum sample volume required, method of urine extraction, potential for contamination of the sample, stability of the analyte of interest, and burden on participants and research team. Collection methods must not introduce contamination or affect the integrity of the sample, should be logistically practical, and should minimize discomfort experienced by the child. Although collection of urine samples from children who are not toilet-trained is more challenging than collection from older toilet-trained children, the vulnerability of younger children to the exposure to and health effects of environmental chemicals makes finding suitable methods a priority. PMID- 19550437 TI - CRTAP mutations in lethal and severe osteogenesis imperfecta: the importance of combining biochemical and molecular genetic analysis. AB - Autosomal recessive lethal and severe osteogenesis imperfecta (OI) is caused by the deficiency of cartilage-associated protein (CRTAP) and prolyl-3-hydroxylase 1 (P3H1) because of CRTAP and LEPRE1 mutations. We analyzed five families in which 10 individuals had a clinical diagnosis of lethal and severe OI with an overmodification of collagen type I on biochemical testing and without a mutation in the collagen type I genes. CRTAP mutations not described earlier were identified in the affected individuals. Although it seems that one important feature of autosomal recessive OI due to CRTAP mutations is the higher consistency of radiological features with OI type II-B/III, differentiation between autosomal dominant and autosomal recessive OI on the basis of clinical, radiological and biochemical investigations proves difficult in the affected individuals reported here. These observations confirm that once a clinical diagnosis of OI has been made in an affected individual, biochemical testing for overmodification of collagen type I should always be combined with molecular genetic analysis of the collagen type I genes. If no mutations in the collagen type I genes are found, additional molecular genetic analysis of the CRTAP and LEPRE1 genes should follow. This approach will allow proper identification of the genetic cause of lethal or severe OI, which is important in providing prenatal diagnosis, preimplantation genetic diagnosis and estimating recurrence risk. PMID- 19550440 TI - Simple models for laser-induced damage and conditioning of potassium dihydrogen phosphate crystals by nanosecond pulses. AB - When potassium dihydrogen phosphate crystals (KH(2)PO(4) or KDP) are illuminated by multi-gigawatt nanosecond pulses, damages may appear in the crystal bulk. One can increase damage resistance through a conditioning that consists in carrying out a laser pre-exposure of the crystal. The present paper addresses the modeling of laser-induced damage and conditioning of KDP crystals. The method is based on heating a distribution of defects, the cooperation of which may lead to a dramatic temperature rise. In a previous investigation [Opt. Express 15, 4557 4576 (2007)], calculations were performed for cases where the heat diffusion was permitted in one and three spatial dimensions, corresponding respectively to planar and point defects. For the sake of completeness, the present study involves the 2D heat diffusion that is associated with linear defects. A comparison to experimental data leads to the conclusion that 1D calculations are the most appropriate for describing the laser-induced damage in KDP. Within this framework, the evolution of the damage density is given as a function of the laser energy density and an in-depth analysis of the results is provided based on simple analytical expressions that can be used for experimental design. Regarding the conditioning, assuming that it is due to a decrease in the defect absorption efficiency, two scenarios associated with various defect natures are proposed and these account for certain of the observed experimental facts. For instance, in order to improve the crystal resistance to damage, one needs to use a conditioning pulse duration shorter than the testing pulse. Also, a conditioning scenario based on the migration of point (atomic-size) defects allows the reproduction of a logarithmic-like evolution of the conditioning gain with respect to the number of laser pre-exposures. Moreover, this study aims at refining the knowledge regarding the precursor defects responsible for the laser induced damage in KDP crystals. Within the presented modeling, the best candidate permitting the reproduction of major experimental facts is comprised of a collection of one-hundred-nanometer structural defects associated with point defects as for instance cracks and couples of oxygen interstitials and vacancies. PMID- 19550441 TI - Compact and low power thermo-optic switch using folded silicon waveguides. AB - By exploiting the small bend radius achievable using high-index-contrast silicon photonic wire waveguides, we demonstrate a new low power thermo-optic switch arranged in a dense, double spiral geometry. Such a design permits the waveguide length to be extended for increased phase shift, without the need for increased heated volume. This provides an effective means to reduce the power consumption of thermo-optic switches, as well as a compact geometry desirable for the development of switch arrays. A low switching power of 6.5 mW was obtained for a spiral-path Mach-Zehnder interferometer device having a 10% - 90% rise time of 14 micros. The switching power is shown to be reduced by more than 5 times compared to a Mach-Zehnder interferometer employing a conventional straight waveguide geometry. PMID- 19550443 TI - Extreme axial optical force in a standing wave achieved by optimized object shape. AB - Standing wave optical trapping offers many useful advantages in comparison to single beam trapping, especially for submicrometer size particles. It provides axial force stronger by several orders of magnitude, much higher axial trap stiffness, and spatial confinement of particles with higher refractive index. Mainly spherical particles are nowadays considered theoretically and trapped experimentally. In this paper we consider prolate objects of cylindrical symmetry with radius periodically modulated along the axial direction and we present a theoretical study of optimized objects shapes resulting in up to tenfold enhancement of the axial optical force in comparison with the original unmodulated object shape. We obtain analytical formulas for the axial optical force acting on low refractive index objects where the light scattering by the object is negligible. Numerical results based on the coupled dipole method are presented for objects with higher refractive indices and they support the previous simplified analytical conclusions. PMID- 19550442 TI - Fluctuations of scattered waves: going beyond the ensemble average. AB - The interaction between coherent waves and random media is a complicated, deterministic process that is usually examined upon ensemble averaging. The result of one realization of the interaction process depends on the specific disorder present in an experimentally controllable interaction volume. We show that this randomness can be quantified and structural information not apparent in the ensemble average can be obtained. We use the information entropy as a viable measure of randomness and we demonstrate that its rate of change provides means for discriminating between media with identical mean characteristics. PMID- 19550446 TI - Predissociation dynamics of the B state of CH3I by femtosecond pump-probe technique. AB - The laser induced predissociation dynamics of the B Rydberg state of CH(3)I following two-photon absorption of a pump pulse was studied with femtosecond pump probe photoelectron imaging coupled with time-resolved mass spectroscopy. The predissociation lifetime was measured to be 1.55 ps induced by the crossing between the B state and the repulsive A-band. Two possible predissociation channels were observed originating from (a) direct coupling between the B state and the repulsive (3)Q(0) state and (b) a second crossing between the (3)Q(0) and (1)Q(1) states after the coupling between the B and (3)Q(0) states, respectively. PMID- 19550445 TI - Dielectric liquid microlens with well-shaped electrode. AB - A dielectric liquid microlens with a well-shaped electrode is demonstrated. The bottom well-shaped electrode and the top planar electrode induce an inhomogeneous electric field when a voltage is applied, which causes the focal length to change. Adaptive microlenses and microlens arrays with well-shaped electrode are fabricated and their performance is evaluated. The bi-convex structure introduces a larger optical power. Compared with common planar-electrodes liquid lenses, this well-shaped electrode not only inhibits drifting of the liquid but also reduces the operating voltage. PMID- 19550447 TI - 45 degree polymer micromirror integration for board-level three-dimensional optical interconnects. AB - We introduce here a simple method of integrating 45 degrees total internal reflection micro-mirrors with polymer optical waveguides by an improved tilted beam photolithography on printed circuit boards to provide surface normal light coupling between waveguides and optoelectronic devices for optical interconnects. De-ionized water is used to couple ultraviolet beam through the waveguide core polymer layer at 45 degrees angle during the photo exposure process. This technique is compatible with PCB manufacturing facility and suitable to large panel board-level manufacturing. The mirror slope is controlled accurately (within +/- 1 degrees) with high repeatability. The insertion loss of an uncoated micro-mirror is measured to be 1.6 dB. PMID- 19550444 TI - A real-time photoacoustic tomography system for small animals. AB - A real-time 512-element photoacoustic tomography system for small animal imaging using a ring ultrasound array has been developed. The system, based upon a 5 MHz transducer array formed along a 50 mm circular aperture, achieves sub-200 micron lateral resolution over a 2 cm disk-shaped region. Corresponding elevation resolutions of 0.6 to 2.5 mm over the central volume enable depth-resolved 3D tomographic imaging with linear translation. Using 8:1 electronic multiplexing, imaging at up to 8 frame/sec is demonstrated for both dynamic phantoms and in vivo mouse and brain samples. The real-time, full 2D tomographic capability of the system paves the way for functional photoacoustic tomographic imaging studies in small animals with sub-second time frame. PMID- 19550448 TI - Numerical analysis of resonant properties of a waveguide structure within a random medium. AB - We propose a simple structure for manipulating resonant conditions in random structures, which is composed of a waveguide structure as a defect region embedded in a random structure. Using the two-dimensional finite-difference time domain method, we examine the resonant properties of localized modes bound in the waveguide. From the results, we confirm that long-lived modes are strongly confined in the waveguide only when the resonant frequency matches the frequency windows in the transmitted intensity spectrum of the surrounding random structure. PMID- 19550450 TI - Linear polarization-discriminatory state inverter fabricated by oblique angle deposition. AB - In this paper, we report a linear polarization-discriminatory state inverter made of three-layer sculpture thin film fabricated by oblique angle deposition technique. The first and third layers are quarter-wave plates of zigzag structure and the middle of them is a circular Bragg reflector of left-handed helical structure. It is found that the normal incidence of P-polarized light on this polarization-discriminatory state inverter becomes the S-polarized light at output, while the incident S-polarized light of wavelength lying in the Bragg regime is reflected. The microstructure of the linear polarization-discriminatory state inverter is also investigated by using a scanning electron microscope. PMID- 19550451 TI - Efficient spectroscopy of single embedded emitters using optical fiber taper waveguides. AB - A technique based on using optical fiber taper waveguides for probing single emitters embedded in thin dielectric membranes is assessed through numerical simulations. For an appropriate membrane geometry, photoluminescence collection efficiencies in excess of 10% are predicted, exceeding the efficiency of standard free-space collection by an order of magnitude. Our results indicate that these fiber taper waveguides offer excellent prospects for performing efficient spectroscopy of single emitters embedded in thin films, such as a single self assembled quantum dot in a semiconductor membrane. PMID- 19550449 TI - Propagation of a partially coherent cosine-Gaussian beam through an ABCD optical system in turbulent atmosphere. AB - The propagation of a partially coherent cosine-Gaussian beam through a paraxial ABCD optical system in turbulent atmosphere is investigated. Analytical expression for the average intensity in the output plane is derived. The presented formula only covers optical systems without inherent apertures, where ABCD are all real-valued. As a special case of the general formula, the analytical formula for the average intensity of a partially coherent cosh Gaussian beam through an ABCD optical system in turbulent atmosphere is also presented, respectively. The properties of the average intensity of the partially coherent cosine-Gaussian beam are investigated with a numerical example, and the dependence of the average intensity distribution on the spatial correlation length of a partially coherent cosine-Gaussian beam is mainly discussed. PMID- 19550452 TI - Fresnel-Gaussian shape invariant for optical ray tracing. AB - We propose a technique for ray tracing, based in the propagation of a Gaussian shape invariant under the Fresnel diffraction integral. The technique uses two driving independent terms to direct the ray and is based on the fact that at any arbitrary distance, the center of the propagated Gaussian beam corresponds to the geometrical projection of the center of the incident beam. We present computer simulations as examples of the use of the technique consisting in the calculation of rays through lenses and optical media where the index of refraction varies as a function of position. PMID- 19550454 TI - Three-dimensional quantitative imaging of retinal and choroidal blood flow velocity using joint Spectral and Time domain Optical Coherence Tomography. AB - Recently, joint Spectral and Time domain Optical Coherence Tomography (joint STdOCT) has been proposed to measure ocular blood flow velocity. Limitations of CCD technology allowed only for two-dimensional imaging at that time. In this paper we demonstrate fast three-dimensional STdOCT based on ultrahigh speed CMOS camera. Proposed method is straightforward, fully automatic and does not require any advanced image processing techniques. Three-dimensional distributions of axial velocity components of the blood in human eye vasculature are presented: in retinal and, for the first time, in choroidal layer. Different factors that affect quality of velocity images are discussed. Additionally, the quantitative measurement allows to observe a new interesting optical phenomenon - random Doppler shift in OCT signals that forms a vascular pattern at the depth of sclera. PMID- 19550453 TI - Theory and experiment of a fiber loop mirror filter of two-stage polarization maintaining fibers and polarization controllers for multiwavelength fiber ring laser. AB - A fiber loop mirror (FLM) filter with two-stage polarization-maintaining fibers (PMFs) and polarization controllers (PCs) is presented. The transmission function of this FLM is calculated in detail by Jones matrix. The wavelength interval depends on both the PMFs and the PCs. The side frequencies can be restrained by choosing appropriate length of the PMFs. Furthermore, an erbium-doped fiber ring laser based on this FLM filter is proposed and demonstrated. Stable single-, double- and triple-wavelength are achieved respectively. The 3 dB line-width is less than 0.03 nm, and the fluctuation of wavelength and peak power is less than 0.05 nm and 0.1 dB in 30 minutes. PMID- 19550455 TI - Aerogel waveplates. AB - Optical transmission measurements were made on 98% porosity silica aerogel samples under various degrees of uniaxial strain. Uniaxially compressed aerogels exhibit large birefringence, proportional to the amount of compression, up to the 15% strain studied. The birefringence is mostly reversible and reproducible through multiple compression-decompression cycles. Our study demonstrates that uniaxially strained high porosity aerogels can be used as tunable waveplates in a broad spectral range. PMID- 19550458 TI - Tunable structures comprising two photonic crystal slabs--optical study in view of multi-analyte enhanced detection. AB - Using finite-difference time-domain method, we characterize the normal-incidence transmission properties of a two slab photonic crystal device in a view of its applications in fluorescence enhancement and multi-analyte detection. Individual slabs consist of a square or a triangular lattice of air holes embedded into a silicon nitride slab. The geometrical parameters are chosen so that the individual slabs operate in a guided resonance regime where strong reflectivity under the normal incidence angle is observed in a broad spectral range. When placed in the close proximity of each other, the two photonic crystal slab system exhibits a narrow Fabry-Perot type transmission peak corresponding to the excitation of a resonant mode in the cavity formed by the two slabs. We then study the effects of the size of the air gap between the two photonic crystal slabs on the spectral position and bandwidth of a resonance transmission peak. Finally, we investigate the electromagnetic energy distributions at the wavelength of a transmission resonance in the double slab photonic crystals. As a final result we demonstrate that this structure can provide electric field enhancement at the slab surface, which can be used for fluorescence enhancement. PMID- 19550456 TI - Long-range surface plasmon-polariton waveguide sensors with a Bragg gratingin the asymmetric double-electrode structure. AB - We propose a Bragg grating resonance sensor based on long-range surface plasmon polaritons (LRSPP) excited on an asymmetric double-electrode waveguide structure. The proposed LRSPP waveguide sensor utilizes spectral resonance of the asymmetric double-electrode structure by adding a Bragg grating layer on the top surface of the metal slab. We have numerically estimated the bulk index resolution and thickness detection limit of a target biomolecule layer under 30 dB total propagation loss. The sub-mum fluidic channel between the two metal layers always experiences a highly confined LRSPP mode excited by end-fire coupling with optical fibers, therefore the proposed LRSPP sensor platform can be applied to a variety of integrated sensor-chip scenarios. PMID- 19550457 TI - Efficient LED light distribution cavities using low loss, angle-selective interference transflectors. AB - Recent advances in solid state light source efficiency and luminance present the technical challenge of distributing light from very small point sources to large areas, with area distribution ratios having orders of magnitude greater than previously addressed. Broad adoption of LEDs in lighting and liquid crystal displays is in part contingent on addressing this fundamental light distribution issue. Here we present new materials based on giant birefringent nanotechnology which address these deficiencies allowing us to guide light in air via a novel light distribution system. Resulting from controlled in-plane and out-of-plane x,y,z refractive indices of adjacent layers, these multilayer interference films possess both angle selective and polarization selective reflectance. The angle selectivity can be tuned in both azimuth and polar angle, relieving a key constraint of prior materials. Our work has been done on a physically large scale enabling demonstration of large light management systems of industrial and practical relevance. PMID- 19550459 TI - 3-D coherence holography using a modified Sagnac radial shearing interferometer with geometric phase shift. AB - A new image reconstruction scheme for coherence holography using a modified Sagnac-type radial shearing interferometer with geometric phase shift is proposed, and the first experimental demonstration of generic Leith-type coherence holography, which reconstructs off-axis 3-D objects with depth information, is presented. The reconstructed image, represented by a coherence function, can be visualized with a controllable magnification, which opens up a new possibility for a coherence imaging microscope. PMID- 19550460 TI - Molecular orientation in self-assembled multilayers measured by second harmonic generation using femtosecond pulses. AB - We present measurements of the optical second-harmonic generation in self assembled multilayer films of PAZO/PAH polymers with the aim to investigate molecular order in the layer-by-layer architecture. The experiments are performed in transmission, using a femtosecond Ti:Sa pulsed laser, which allows a more accurate determination of the amplitude of the second harmonic signal, without interference fringes usually present in nanosecond experiments. We found that the first bilayer, in contact with the substrate, presents a broad distribution of the orientation of the molecules, while the addition of successive bilayers (up to 12) produces ordering of the molecules with a small tilt angle respect to the surface normal. This result, together with the monotonic increment of the second harmonic signal with the number of layers indicates that the molecules grow orderly assembled in these films. PMID- 19550461 TI - Stable, continuous-wave, intracavity, optical parametric oscillator pumped by a semiconductor disk laser (VECSEL). AB - We report relaxation oscillation free, true continuous-wave operation of a singly resonant, intracavity optical parametric oscillator (OPO) based upon periodically poled, MgO-doped LiNbO3 and pumped internal to the cavity of a compact, optically excited semiconductor disk laser (or VECSEL). The very short upper-laser-state lifetime of this laser gain medium, coupled with the enhancing effect of the high finesse pump laser cavity in which the OPO is located, enables a low threshold, high efficiency intracavity device to be operated free of relaxation oscillations in continuous-wave mode. By optimizing for low-power operation, parametric threshold was achieved at a diode-laser power of only 1.4 W. At 8.5 W of diode laser power, 205 mW of idler power was extracted, indicating a total down converted power of 1.25 W, and hence a down-conversion efficiency of 83%. PMID- 19550462 TI - Multiscale lens design. AB - While lenses of aperture less than 1000lambda frequently form images with pixel counts approaching the space-bandwidth limit, only heroic designs approach the theoretical information capacity at larger scales. We propose to use the field processing capabilities of small-scale secondary lens arrays to correct aberrations due to larger scale objective lenses, with an ultimate goal of achieving diffraction-limited imaging for apertures greater than 10,000lambda .We present an example optical design using an 8 mm entrance pupil capable of resolving 20 megapixels. PMID- 19550463 TI - High amplification and low noise achieved by a double-stage non-collinear Brillouin amplifier. AB - We report a double-stage non-collinear Brillouin amplifier structure with high amplification and low noise, achieving an energy amplification of 6 x 10(11) and a signal-to-noise ratio of 10(3) for an input signal of 5.5 x 10(-14)J in the regime above the pump's stimulated Brillouin scattering threshold. The signal of the first-stage amplifier is efficiently amplified and separated from the noise output. The saturation amplification with noise suppressing is implemented in the second stage. The design principles of system parameters such as the intersection angle between the pump and signal beams, the pump energy, and the beam diameter are given. PMID- 19550465 TI - Manipulating spatial qudit states with programmable optical devices. AB - The study of how to generate high-dimensional quantum states (qudits) is justified by the advantages that they can bring for the field of quantum information. However, to have some real practical potential for quantum communication, these states must be also of simple manipulation. Spatial qudits states, which are generated by engineering the transverse momentum of the parametric down-converted photons, have been until now considered of hard manipulation. Nevertheless, we show in this work a simple technique for modifying these states. This technique is based on the use of programmable diffractive optical devices, that can act as spatial light modulators, to define the Hilbert space of these photons instead of pre-fabricated multi-slits. PMID- 19550464 TI - Improved phase sensitivity in spectral domain phase microscopy using line-field illumination and self phase-referencing. AB - We report a quantitative phase microscope based on spectral domain optical coherence tomography and line-field illumination. The line illumination allows self phase-referencing method to reject common-mode phase noise. The quantitative phase microscope also features a separate reference arm, permitting the use of high numerical aperture (NA > 1) microscope objectives for high resolution phase measurement at multiple points along the line of illumination. We demonstrate that the path-length sensitivity of the instrument can be as good as 41 pm/square root of Hz, which makes it suitable for nanometer scale study of cell motility. We present the detection of natural motions of cell surface and two-dimensional surface profiling of a HeLa cell. PMID- 19550467 TI - Multistage optical Stark decelerator for a pulsed supersonic beam with a quasi-cw optical lattice. AB - We propose a new scheme to realize a multistage optical Stark deceleration for a supersonic molecular beam using a time-varying red-detuned quasi-cw optical lattice with a length of up to 10 mm. We analyze the motion of the slowed molecules inside the optical decelerator, and study the dependences of the velocity of the slowed molecular packet on the synchronous phase angle and the number of the deceleration stages (i.e., the number of the optical-lattice cells) by using Monte-Carlo method. Our study shows that the proposed optical Stark decelerator cannot only efficiently slow a pulsed supersonic beam from 230 m/s to zero (standstill), but also obtain an ultracold molecular packet with a temperature of sub-mK due to bunching effect in the multistage optical Stark decelerator, which can be trapped in the optical lattice by rapidly turning off the modulation signal of the lattice. Also, we compare the decelerated results of our multistage optical Stark decelerator with a single-stage optical one, and find that our scheme cannot only obtain a colder molecular packet under the same molecular-beam parameters and deceleration conditions, but also be directly used to trap the slowed cold molecules after the deceleration, while don't need to use another molecular trap. PMID- 19550466 TI - Simultaneous reconstruction of phase and amplitude contrast from a single holographic record. AB - We present a reconstruction technique for simultaneous retrieval of absorption and phase shifting properties of an object recorded by in-line holography. The routine is experimentally tested by applying it to optical holograms of a pure phase respectively a pure amplitude object of micrometer dimensions that has been machined into a glass-plate using a focused ion beam. The method has also been applied to previously published electron holograms of single DNA molecules. PMID- 19550468 TI - Characteristics of a multi-mode interference device based on Ti:LiNbO3 channel waveguide. AB - We have analyzed the multi-mode interference effect depending on the wavelength and the polarization states of input beam in a multi-mode Ti:LiNbO(3) waveguide at about 1300 nm region. The transmitted optical signal of a Ti:LiNbO(3) waveguide shows the periodic oscillation as a function of input wavelength. The measured average periodicity of the oscillation in TM and TE polarization beams were about 18 nm and 48 nm, respectively. Actually, the periodicity is determined by the refractive index difference between the two modes (fundamental and first modes). Therefore, we have explained the experimental results with the theoretical calculations which are derived from a quasi-analytical technique based on the effective-refractive- index method and the equation of coupling length determined by the mode phase factor in the multi-mode waveguide. PMID- 19550469 TI - 227-fs pulses from a mode-locked Yb:LuScO3 thin disk laser. AB - We report on the first mode-locked thin disk laser based on Yb:LuScO(3). This new mixed gain material combines the emission peaks of two sesquioxides, leading to a gain bandwidth of more than 20 nm. We achieve 7.2 W average output power in 227 fs pulses, which is shorter than for any previous ultrafast thin disk laser. The output power was limited by a growth defect near the center of the thin disk. PMID- 19550470 TI - A reflective microring notch filter and sensor. AB - We present a new design of wavelength selective reflector composed of a Y junction and a singly coupled microring resonator, and demonstrate its biochemical sensing applications with a prototype device. In contrast with other reflectors like distributed Bragg reflectors, this device acts as notch filter at its reflection port. One promising application of the device is for remote sensing of harsh or inaccessible site, where only one optical fiber is required to transmit the input and reflected light signal over a long distance. The design can also be used to make microring cavity lasers. PMID- 19550471 TI - Spectroscopy of a tapered-fiber photonic crystal waveguide coupler. AB - A spectroscopic technique based on a full four port modeling and measurement is developed and utilized to study a tapered- fiber photonic crystal waveguide coupler. The coupler is made by directly situating the tapered fiber on the defect region of a silicon membrane photonic crystal waveguide. The waveguide is lithographically terminated resulting a Fabry- Perot cavity. It turns out that the line-shape of the resonances is not merely a Lorentzian but can be constructed from that of the unloaded waveguide resonator. By knowing how the resonances broaden the experimental data is then fit and the coupling efficiency is extracted for the entire spectrum. PMID- 19550472 TI - Entanglement swapping using telecom-band photons generated in fibers. AB - We report the first entanglement swapping experiment using entangled photon-pair sources based on spontaneous four-wave mixing (SFWM). The 1.5-microm band entangled photon pairs generated by SFWM in two independent 500-m dispersion shifted fibers exhibited quantum interference, thanks to the negligible walk-off between the pump and photon pairs. The use of 500-MHz gated-mode InGaAs/InP avalanche photodiodes based on the sine-wave gating technique increased the fourfold coincidence rate. As a result, the formation of an entanglement between photons from independent sources was successfully observed. PMID- 19550473 TI - Absorption switches in metal-dielectric-metal plasmonic waveguides. AB - We theoretically investigate the properties of absorption switches for metal dielectric-metal (MDM) plasmonic waveguides. We show that a MDM waveguide directly coupled to a cavity filled with an active material with tunable absorption coefficient can act as an absorption switch, in which the on/off states correspond to the absence/presence of optical pumping. We also show that a MDM plasmonic waveguide side-coupled to a cavity filled with an active material can operate as an absorption switch, in which the on/off states correspond to the presence/absence of pumping. For a specific modulation depth, the side-coupled cavity switch results in more compact designs compared to the direct-coupled cavity switch. Variations in the imaginary part of the refractive index of the material filling the cavity of Deltakappa = 0.01 (Deltakappa = 0.15) result in approximately 60% (approximately 99%) modulation depth. The properties of both switches can be accurately described using transmission line theory. PMID- 19550474 TI - Effect of metallic surface on electric dipole and magnetic dipole emission transitions in Eu3+ doped polymeric film. AB - Spontaneous emission of Eu(3+) ions is studied in thin organic films deposited onto several different substrates. It has been demonstrated that the presence of a metallic surface in close vicinity to emitting Eu(3+) ions causes modifications of their spontaneous emission spectra, in particular, the change in the relative strengths of magnetic-dipole and electric-dipole transitions. The character and the magnitude of the effect depend on the polarization and the observation angle. The experimental data are discussed in terms of modification of transition probabilities and account for the interference between directly emitted and reflected light waves. PMID- 19550476 TI - High-precision open-loop adaptive optics system based on LC-SLM. AB - Used as a wavefront corrector, a liquid crystal spatial modulator (LC-SLM) has good repeatability and linearity, which are essential for open-loop adaptive optics, and the open-loop optical system can increase the light energy efficiency by a factor of two for the LC-SLM and improve the system bandwidth. In order to test the performance of the LC-SLM in open-loop correction, an indoor closed-loop configuration optical system is constructed on the open-loop control method. With this method, it is demonstrated that the residual error after open-loop correction could be smaller than 0.08lambda (RMS: root mean square value) if the initial wavefront aberration is below 2.5lambda (RMS), and the repeatability error of open-loop correction is smaller than 0.01lambda (RMS). PMID- 19550477 TI - Origin of light-deflection in lithium niobate and lithium tantalate under electric field. AB - The deflection of light reported by Muller et al. in lithium niobate [Appl. Phys. B 78, 367-370] and lithium tantalate [Appl. Optics 43 (34), 6344-6347] under electric field originates from refraction at domain-walls, like in ferroelastics. In ferroelectrics the optical discontinuity takes place at domain-walls as a consequence of the electro-optic effect. The theoretical deflection angle calculated from Snell's law is proportional to the square root of the electric field and matches the experimental results reported by Muller et al. for lithium niobate. The finite domain-wall thickness mentioned by the authors is not involved in the deflection phenomenon. PMID- 19550479 TI - Optical magnetic plasma in artificial flowers. AB - We report the design of an artificial flower-like structure that supports a magnetic plasma in the optical domain. The structure is composed of alternating "petals" of conventional dielectrics (epsilon > 0) and plasmonic materials (Re(epsilon ) < 0). The induced effective magnetic current on such a structure possesses a phase lag with respect to the incident TE-mode magnetic field, similar to the phase lag between the induced electric current and the incident TM mode electric field on a metal wire. An analogy is thus drawn with an artificial electric plasma composed of metal wires driven by a radio frequency excitation. The effective medium of an array of flowers has a negative permeability within a certain wavelength range, thus behaving as a magnetic plasma. PMID- 19550478 TI - High temporal resolution OCT using image-based retrospective gating. AB - High temporal resolution OCT imaging is very advantageous for analyzing cardiac mechanics in the developing embryonic heart of small animals. An image-based retrospective gating technique is presented to increase the effective temporal resolution of an OCT system and to allow visualization of systolic dynamics in 3D. The gating technique employs image similarity measures for rearranging asynchronously acquired input data consisting of a time series of 2D images at each z position along the heart volume, to produce a time sequence of 3D volumes of the beating heart. The study includes a novel robust validation technique, which quantitatively evaluates the accuracy of the gating technique, in addition to visual evaluations by 2D multiplanar reformatting (MPR) and 3D volume rendering. The retrospective gating and validation is demonstrated on a stage 14 embryonic quail heart data set. Using the validation scheme, it is shown that the gating is accurate within a standard deviation of 4.7 ms, which is an order of magnitude shorter than the time interval during which systolic contraction (approximately 50 ms) occurs in the developing embryo. This gating method has allowed, for the first time, clear visualization of systolic dynamics of the looping embryonic heart in 3D. PMID- 19550480 TI - Improved color rendering and luminous efficacy in phosphor-converted white light emitting diodes by use of dual-blue emitting active regions. AB - Conventional white-light sources suffer from a fundamental trade-off between color rendering index and the luminous efficacy; increasing one generally comes at the expense of the other. We demonstrate through simulation that dual wavelength blue-emitting active regions in phosphor-converted white light sources maximize the output luminous flux while significantly increasing the color rendering ability. Our results indicate that such improvements can be achieved over a broad range of correlated color temperatures. PMID- 19550481 TI - Which is the most power-efficient modulation format in optical links? AB - By exploiting the electromagnetic wave's four-dimensional signal space, we find that for the additive white Gaussian noise channel, the modulation format with best sensitivity to be an 8-level format with 1.76 dB asymptotic gain over BPSK, for uncoded optical transmission with coherent detection. Low-complexity modulators are presented for the format, as well as an interpretation in terms of quantum-limited sensitivity. PMID- 19550482 TI - Mode-locked InGaAs-AlGaAs disk laser generating sub-200-fs pulses, pulse picking and amplification by a tapered diode amplifier. AB - Almost chirp-free pulses with a duration of 190 fs were achieved from a mode locked semiconductor disk laser (SDL) emitting at approximately 1045 nm. Pulse shaping was different from the soliton-like mode-locking process known from lasers using dielectric gain media; passive amplitude modulation provided by a fast saturable absorber was essential. The spectrum of the absorber had to be matched to the gain spectrum within a few nm. A tapered diode amplifier was demonstrated to be a device for both picking and amplifying SDL pulses. The pulse repetition rate of the SDL output was reduced from 3 GHz to 47 MHz. PMID- 19550483 TI - Distributed nonlinear fiber chirped-pulse amplifier system. AB - A two-stage fiber-based femtosecond amplification system is presented, based on chirped-pulse amplification in highly nonlinear regime. The amount of self-phase modulation is separately adjusted in each stage selecting the proper stretching ratio in order to compensate gain narrowing. Analytical design rules are validated using numerical simulations. Our experimental implementation leads to the generation of high temporal quality 20 microJ 202 fs pulses at repetition rate of 200 kHz, a record duration at this energy level. PMID- 19550484 TI - Spectral artifacts from non-uniform samples analyzed by terahertz time-domain spectroscopy. AB - We report the impact of the spatial coherence distortion on the measured absorption spectra and the identification of materials analyzed by terahertz time domain spectroscopy. It is shown that the deformation of the terahertz beam wave front can result into the overestimation of the electromagnetic absorption, the generation of artificial absorption peaks and even to the disappearance of characteristic absorption peaks. Obtaining clear absorption spectra without artifacts is crucial for applications based on terahertz imaging and spectroscopy. PMID- 19550485 TI - Clinical system for non-invasive in situ monitoring of gases in the human paranasal sinuses. AB - We present a portable system for non-invasive, simultaneous sensing of molecular oxygen (O(2)) and water vapor (H(2)O) in the human paranasal cavities. The system is based on high-resolution tunable diode laser spectroscopy (TDLAS) and digital wavelength modulation spectroscopy (dWMS). Since optical interference and non ideal tuning of the diode lasers render signal processing complex, we focus on Fourier analysis of dWMS signals and procedures for removal of background signals. Clinical data are presented, and exhibit a significant improvement in signal-to-noise with respect to earlier work. The in situ detection limit, in terms of absorption fraction, is about 5x10(-5) for oxygen and 5x10(-4) for water vapor, but varies between patients due to differences in light attenuation. In addition, we discuss the use of water vapor as a reference in quantification of in situ oxygen concentration in detail. In particular, light propagation aspects are investigated by employing photon time-of-flight spectroscopy. PMID- 19550486 TI - Goos-Hanchen shift for a rough metallic mirror. AB - We investigate experimentally the dependence of the Goos-Hanchen shift on the surface properties of an air-metal interface. The shift depends on the microscopic roughness of the metal surface but it is insensitive to the large scale variations associated with surface non-flatness. Both an effective medium model of roughness and the Rayleigh-Rice theory of scattering are used to interpret the observed phenomenon. PMID- 19550487 TI - Investigation on mechanism of multiple holographic recording with uniform diffraction efficiency in photopolymers. AB - In this paper, our simplified model for grating formation in photopolymers is extended to describe the effects of uniform post-exposure on an existing hologram, and further to present a new model of holographic multiplexing for calculating the exposure schedule for multiplexed gratings with uniform diffraction efficiency. It is experimentally verified that the refractive-index modulation of the existing gratings continues to increase when a current grating is being recorded. Twenty gratings were multiplexed using the exposure schedule calculated with the uniform post-exposure model, and comparison with the result from traditional method confirmed the validity of this model. PMID- 19550488 TI - Endface reflectivities of optical nanowires. AB - Endface reflectivities (ERs) of optical nanowires are investigated using three dimensional finite-difference time-domain simulations. Typical ERs of both free standing and substrate-supported silica, tellurite, PMMA and semiconductor nanowires or nanofibers are obtained. Unlike in conventional waveguides such as optical fibers, ERs of nanowires are usually considerably lower when operated in single mode. Dependences of ER on the diameter and the refractive index of the nanowire, and the wavelength of the guided light are also investigated. These results are helpful for estimating and understanding ERs in optical nanowires with diameters close to or smaller than the wavelengths of the light, and may offer valuable references for practical applications such as nanowire or nanofiber-based resonators and lasers. PMID- 19550489 TI - Midinfrared absorption measured at a lambda/400 resolution with an atomic force microscope. AB - Midinfrared absorption can be locally measured using a detection combining an atomic force microscope and a pulsed excitation. This is illustrated for the midinfrared bulk GaAs phonon absorption and for the midinfrared absorption of thin SiO(2) microdisks. We show that the signal given by the cantilever oscillation amplitude of the atomic force microscope follows the spectral dependence of the bulk material absorption. The absorption spatial resolution achieved with microdisks is around 50 nanometer for an optical excitation around 22 micrometer wavelength. PMID- 19550490 TI - A finite element beam propagation method for simulation of liquid crystal devices. AB - An efficient full-vectorial finite element beam propagation method is presented that uses higher order vector elements to calculate the wide angle propagation of an optical field through inhomogeneous, anisotropic optical materials such as liquid crystals. The full dielectric permittivity tensor is considered in solving Maxwell's equations. The wide applicability of the method is illustrated with different examples: the propagation of a laser beam in a uniaxial medium, the tunability of a directional coupler based on liquid crystals and the near-field diffraction of a plane wave in a structure containing micrometer scale variations in the transverse refractive index, similar to the pixels of a spatial light modulator. PMID- 19550491 TI - Interaction of radially polarized focused light with a prolate spheroidal nanoparticle. AB - The interaction of a nanoparticle with light is affected by nanoparticle geometry and composition, as well as by focused beam parameters, such as the polarization and numerical aperture of the beam. The interaction of a radially focused beam with a prolate spheroidal nanoparticle is particularly important because it has the potential to produce strong near-field electromagnetic radiation. Strong and tightly localized longitudinal components of a radially polarized focused beam can excite strong plasmon modes on elongated nanoparticles such as prolate spheroids. In this study, near field radiation from a prolate spheriodal nanoparticle is investigated when it is illuminated with a radially polarized focused beam of light. Near-field radiation from the nanoparticle is investigated in the absence and presence of metallic layers. It is shown that the interaction of a radially polarized focused beam with a prolate spheroidal nanoparticle can be enhanced by creating images of monopole charges using metallic layers. In addition, it is also observed that the presence of a metallic layer shifts the resonance of the prolate spheroid toward longer wavelengths. Dipole, quadruple, and off resonance field distributions for particles with different sizes and aspect ratios are presented when they are illuminated with a radially focused beam of light. PMID- 19550492 TI - On the normalization of scintillation autocovariance for generalized SCIDAR. AB - The Generalized SCIDAR (Scintillation Detection and Ranging) technique consists in the computation of the mean autocorrelation of double-star scintillation images taken on a virtual plane located a few kilo-meters below the telescope pupil. This autocorrelation is normalized by the autocorrelation of the mean image. Johnston et al. in 2002 pointed out that this normalization leads to an inexact estimate of the optical-turbulence strength C(2)(N). Those authors restricted their analysis to turbulence at ground level. Here we generalize that study by calculating analytically the error induced by that normalization, for a turbulent layer at any altitude. An exact expression is given for any telescope pupil shape and an approximate simple formula is provided for a full circular pupil. We show that the effect of the inexact normalization is to overestimate the C(2)(N) values. The error is larger for higher turbulent layers, smaller telescopes, longer distances of the analysis plane from the pupil, wider double star separations, and larger differences of stellar magnitudes. Depending on the observational parameters and the turbulence altitude, the relative error can take values from zero up to a factor of 4, in which case the real C(2)(N) value is only 0.2 times the erroneous one. Our results can be applied to correct the C(2)(N) profiles that have been measured using the Generalized SCIDAR technique. PMID- 19550493 TI - All-fiber-based optical frequency generation from an Er-doped fiber femtosecond laser. AB - Generating precise optical frequencies with a functional power is necessary in many fields of science and technology. Here we demonstrate an all-fiber-based apparatus built to generate near-infrared frequencies directly from an Er-doped fiber femtosecond laser. In our apparatus, only a single resonance mode is extracted at a time on demand via a composite fiber filter comprised of a Fabry Perot etalon with a Bragg grating. The extracted mode having weak 40 nW power is amplified to 20 mW by means of optical injection locking to a distributed feedback laser diode under phase-stabilization control. The amplified final output signal yields a frequency stability of 2 parts in 10(15) at 10 s averaging with a narrow linewidth of less than 1 Hz. This apparatus is precise and immune to environmental disturbance, thereby being well suited to on-site near-infrared applications of frequency calibration, spectroscopy, and optical clocks. PMID- 19550494 TI - Spatio-temporal Hotelling observer for signal detection from image sequences. AB - Detection of signals in noisy images is necessary in many applications, including astronomy and medical imaging. The optimal linear observer for performing a detection task, called the Hotelling observer in the medical literature, can be regarded as a generalization of the familiar prewhitening matched filter. Performance on the detection task is limited by randomness in the image data, which stems from randomness in the object, randomness in the imaging system, and randomness in the detector outputs due to photon and readout noise, and the Hotelling observer accounts for all of these effects in an optimal way. If multiple temporal frames of images are acquired, the resulting data set is a spatio-temporal random process, and the Hotelling observer becomes a spatio temporal linear operator. This paper discusses the theory of the spatio-temporal Hotelling observer and estimation of the required spatio-temporal covariance matrices. It also presents a parallel implementation of the observer on a cluster of Sony PLAYSTATION 3 gaming consoles. As an example, we consider the use of the spatio-temporal Hotelling observer for exoplanet detection. PMID- 19550495 TI - Multiple path length dual polarization interferometry. AB - An optical sensor for quantitative analysis of ultrathin films and adsorbed layers is described. Quantification of both layer thickness and refractive index (density) can be made for in situ and ex-situ coated films. With the use of two polarizations, in situ measurements are made via one path length in a young's interferometer arrangement while ex-situ measurements use multiple path lengths. The multiple path length young's interferometer arrangement is embodied in a solid state waveguide configuration called the multiple path length dual polarization interferometer (MPL-DPI). The technique is demonstrated with ultrathin layers of poly(methylmethacrylate) and human serum albumin. PMID- 19550496 TI - Femtosecond third-order optical nonlinearity of BiFeO3. AB - We report both linear and nonlinear optical properties of a ferroelectric thin film of polycrystalline BiFeO(3) deposited on a quartz substrate. The linear refraction index and absorption coefficient of the film are determined as a function of light wavelength by optical transmittance measurements. By performing Z-scan experiments with femtosecond laser pulses at a wavelength of 780 nm, the third-order nonlinear refraction index and two-photon absorption (2PA) coefficient are measured to be 1.5 x 10(-4) cm(2)/GW and 16 cm/GW, respectively. The results indicate that the thin film of polycrystalline BiFeO(3) is a promising candidate for applications in nonlinear photonic devices. PMID- 19550498 TI - High-speed optical switch fabrics with large port count. AB - We report a novel architecture that can be used to construct optical switch fabrics with very high port count and nanoseconds switching speed. It is well known that optical switch fabrics with very fast switching time and high port count are challenging to realize. Currently, one of the most promising solutions is based on a combination of wavelength-tunable lasers and the arrayed waveguide grating router (AWGR). To scale up the number of ports in such switches, a direct method is to use AWGRs with a high channel count. However, such AWGRs introduce very large crosstalk noise due to the close wavelength channel spacing. In this paper, we propose an architecture for realizing a high-port count optical switch fabric using a combination of low-port count AWGRs, optical ON-OFF gates and WDM couplers. Using this new methodology, we constructed a proof-of concept experiment to demonstrate the feasibility of a 256 x 256 optical switch fabric. To our knowledge, this port count is the highest ever reported for switch fabrics of this type. PMID- 19550497 TI - Implementation of quantum state tomography for time-bin entangled photon pairs. AB - Quantum state tomography (QST) is an important method for evaluating the quality of entangled photon pairs, and has been widely used to measure polarization entanglement. However, QST has not been applied to time-bin entanglement, which is a type of entanglement suitable for fiber transmission. In this paper, we clarify the way to implement QST on time-bin entangled photon pairs using a 1-bit delayed interferometer. We also provide experimental results for a demonstration of QST for time-bin entangled photon pairs generated using spontaneous four-wave mixing in a dispersion shifted fiber. PMID- 19550499 TI - Liquid-crystal variable-focus lenses with a spatially-distributed tilt angles. AB - A pretilt angle controlling method by the density of rubbings using a tiny stylus is proposed. The control of the surface pretilt angle is achieved by rubbing a side-chain type polyimide film for a homeotropic alignment. Smooth liquid crystal (LC) director distribution in the bulk layer is successfully obtained even though the rough surface orientation. This approach is applied to LC cylindrical and rectangular lenses with a variable-focusing function. The distribution profile of the rubbing pitch (the reciprocal of the rubbing density) for small aberration is determined to be quadratic. The variable focusing function is successfully achieved in the LC rectangular lens, and the voltage dependence of the focal length is tried to be explained by the LC molecular reorientation behavior. PMID- 19550501 TI - Wave-aberration control with a liquid crystal on silicon (LCOS) spatial phase modulator. AB - Liquid crystal on Silicon (LCOS) spatial phase modulators offer enhanced possibilities for adaptive optics applications in terms of response velocity and fidelity. Unlike deformable mirrors, they present a capability for reproducing discontinuous phase profiles. This ability also allows an increase in the effective stroke of the device by means of phase wrapping. The latter is only limited by the diffraction related effects that become noticeable as the number of phase cycles increase. In this work we estimated the ranges of generation of the Zernike polynomials as a means for characterizing the performance of the device. Sets of images systematically degraded with the different Zernike polynomials generated using a LCOS phase modulator have been recorded and compared with their theoretical digital counterparts. For each Zernike mode, we have found that image degradation reaches a limit for a certain coefficient value; further increase in the aberration amount has no additional effect in image quality. This behavior is attributed to the intensification of the 0-order diffraction. These results have allowed determining the usable limits of the phase modulator virtually free from diffraction artifacts. The results are particularly important for visual simulation and ophthalmic testing applications, although they are equally interesting for any adaptive optics application with liquid crystal based devices. PMID- 19550500 TI - Passive mode-locking of a Tm-doped bulk laser near 2 microm using a carbon nanotube saturable absorber. AB - Stable and self-starting mode-locking of a Tm:KLu(WO(4))(2) crystal laser is demonstrated using a transmission-type single-walled carbon nanotube (SWCNT) based saturable absorber (SA). These experiments in the 2 microm regime utilize the E11 transition of the SWCNTs for nonlinear saturable absorption. The recovery time of the SWCNT-SA is measured by pump-probe measurements as approximately 1.2 ps. The mode-locked laser delivers approximately 10 ps pulses near 1.95 microm with a maximum output power of up to 240 mW at 126 MHz repetition rate. PMID- 19550502 TI - Dependence of resonant light transmission properties of a subwavelength slit on structural parameters. AB - We perform a systematic study of the resonant transmission of visible and near infrared (NIR) light through a single subwavelength slit in a gold film when the parameters defining the structure are varied. We further examine the optical properties of a related nanostructure, a cross with subwavelength sized features. Focused ion beam (FIB) milling was used to fabricate nanoslits and crosses with linewidths ranging from 26 nm to 85 nm. The dimensions of the structure are found to affect strongly the transmittance spectrum. For example, as the slit becomes narrower the resonance is observed to both sharpen and shift significantly. Our observations are in good agreement with our earlier numerical calculations on the optical properties of nanoslits. PMID- 19550503 TI - Realizing near-perfect absorption at visible frequencies. AB - Sub-wavelength hole array (SHA) combined with thick metal layer (TML) is shown to have simultaneous suppressed transmission and reflection, resulting near-perfect absorption. Unlike the simultaneous electric and magnetic resonances in electric ring resonator and cut wire [PRL, 100, 207402 (2008)], such behavior results from strong anti-symmetric surface plasmons coupling supported by SHA and TML. The polarization-free characteristic permits to construct an ideal absorber for some practical applications in turbid backgrounds. PMID- 19550504 TI - Ultrafast all-optical modulation in silicon-based nanoplasmonic devices. AB - A five-layer silicon-based nanoplasmonic waveguiding structure is proposed for ultrafast all-optical modulation and switching applications. Ultrafast nonlinear phase and amplitude modulation is achieved via photo-generated free carrier dynamics in ion-implanted silicon using above-bandgap femtosecond pump pulses. Both an analytical model and rigorous numerical simulations of the structures have shown that a switching time of 5 ps and an on-off contrast of 35 dB can be achieved in these devices. PMID- 19550505 TI - High-resolution computed tomography with a compact soft x-ray microscope. AB - Computed tomography based on high-resolution soft x-ray microscopy utilizes the natural contrast for biological specimens provided by the water window (lambda = 2.4 - 4.4 nm) and the high resolving power of zone plate objectives. It is capable of revealing the 3D structure of biological specimens at sub-visible microscopic resolution. To date, the technique has only been available at synchrotron-based microscopes, which limits the researchers access. In the present paper we demonstrate high-resolution soft x-ray tomography with a laboratory zone-plate-based soft x-ray microscope. The specimen, a diatom mounted on a glass capillary, was reconstructed from a tilt series of 53 images covering 180 degrees using a filtered back projection algorithm. The resolution of the tomogram was estimated to a half period of 140 nm using a differential-phase residual method. Cryo-fixation, increased source brightness and extended-depth-of focus objectives are important for pushing the resolution of compact systems for biological samples. PMID- 19550506 TI - Optimized grating coupler with fully etched slots. AB - A grating coupler with fully etched slots is optimized for fiber coupling into SOI slab waveguides. Such coupler can be produced in one lithography step together with other SOI components. Theoretical maximal coupling efficiency of 49% is demonstrated with a 3 dB bandwidth of 35 nm. Strong reflection from the fully etched grating was avoided through an antireflection interface. Constructive interference is used to decrease radiation into the substrate and the filling factor is optimized for optimal power coupling into the fiber mode. It was also demonstrated, that the chirped grating approach is inapplicable for fully etched gratings. PMID- 19550507 TI - Pico-strain multiplexed fiber optic sensor array operating down to infra-sonic frequencies. AB - An integrated sensor system is presented which displays passive long range operation to 100 km at pico-strain (pepsilon) sensitivity to low frequencies (4 Hz) in wavelength division multiplexed operation with negligible cross-talk (better than -75 dB). This has been achieved by pre-stabilizing and multiplexing all interrogation lasers for the sensor array to a single optical frequency reference. This single frequency reference allows each laser to be locked to an arbitrary wavelength and independently tuned, while maintaining suppression of laser frequency noise. With appropriate packaging, such a multiplexed strain sensing system can form the core of a low frequency accelerometer or hydrophone array. PMID- 19550508 TI - Detection of acoustic pressure with hollow-core photonic bandgap fiber. AB - We report the results of our preliminary investigation on the use of hollow-core photonic bandgap fiber (HC-PBF) for hydrophone application. The response of the commercial HC-1550-02 fiber to acoustic pressure, in terms of normalized responsivity (NR), is measured to be - 334.4 dB re 1 microPa(-1). This agrees well with the theoretically predicated value of -331.6 dB re 1 microPa(-1) and is about 15 dB higher than that of the conventional fiber (HNSM-155). With straightforward fiber structure modifications (thinner outer silica cladding and higher air-filling ratio of inner microstructured cladding), the NR could be further enhanced to - 310 dB re 1 microPa(-1). PMID- 19550509 TI - Suppression of aliasing in multi-sensor scanning absolute profile measurement. AB - The task of anti-aliasing in absolute profile measurement by multi-sensor scanning techniques is considered. Simulation results are presented which demonstrate that aliasing can be highly reduced by a suitable choice of the scanning steps. The simulation results were confirmed by results obtained for interferometric measurements (Nyquist frequency 1/646 microm(-1)) on a specifically designed chirp specimen with sinusoidal waves of amplitude 100 nm and wavelengths from 2.5 mm down to 19 microm. PMID- 19550510 TI - Lasing in metal-insulator-metal sub-wavelength plasmonic waveguides. AB - We demonstrate lasing in Metal-Insulator-Metal (MIM) waveguides filled with electrically pumped semiconductor cores, with core width dimensions below the diffraction limit. Furthermore these waveguides propagate a transverse magnetic (TM0) or so called gap plasmon mode [1-4]. Hence we show that losses in sub wavelength MIM waveguides can be overcome to create small plasmon mode lasers at wavelengths near 1500 nm. We also give results showing room temperature lasing in MIM waveguides, with approximately 310 nm wide semiconductor cores which propagate a transverse electric mode. PMID- 19550511 TI - Quadrupole-dipole transform based on optical near-field interactions in engineered nanostructures. AB - Nanophotonics has the potential to provide novel devices and systems with unique functions based on optical near-field interactions. Here we experimentally demonstrate, for the first time, what we call a quadrupole-dipole transform achieved by optical near-field interactions between engineered nanostructures. We describe its principles, the nanostructure design, fabrication of one- and two layer gold nanostructures, an experimental demonstration, and optical characterization and analysis. PMID- 19550512 TI - Compression of ultrashort laser pulses in planar hollow waveguides: a stability analysis. AB - We investigate compression of ultrashort laser pulses by nonlinear propagation in gas-filled planar hollow waveguides, using (3+1)- dimensional numerical simulations. In this geometry, the laser beam is guided with a fixed size in one transverse dimension, generating significant spectral broadening, while it propagates freely in the other, allowing for energy up-scalability. In this respect the concept outperforms compression techniques based on hollow core fibers or filamentation. Small-scale self-focusing is a crucial consideration, which introduces mode deterioration and finally break-up in multiple filaments. The simulation results, which match well with initial experiments, provide important guidelines for scaling the few-cycle pulse generation to higher energies. Pulse compression down to few-cycle duration with energies up to 100 mJ levels should be possible. PMID- 19550513 TI - Average intensity and spreading of an elegant Hermite-Gaussian beam in turbulent atmosphere. AB - The propagation of an elegant Hermite-Gaussian beam (EHGB) in turbulent atmosphere is investigated. Analytical propagation formulae for the average intensity and effective beam size of an EHGB in turbulent atmosphere are derived based on the extended Huygens-Fresnel integral. The corresponding results of a standard Hermite-Gaussian beam (SHGB) in turbulent atmosphere are also derived for the convenience of comparison. The intensity and spreading properties of EHGBs and SHGBs in turbulent atmosphere are studied numerically and comparatively. It is found that the propagation properties of EHGBs and SHGBs are much different from their properties in free space, and the EHGB and SHGB with higher orders are less affected by the turbulence. What's more, the SHGB spreads more rapidly than the EHGB in turbulent atmosphere under the same conditions. Our results will be useful in long-distance free-space optical communications. PMID- 19550514 TI - Monitoring technique for waveform distortion of 160 Gb/s signal by prescaled clock tone detection using EA modulator. AB - In order to monitor quality of ultra high bit-rate optical signals in a future optical network, such as 160 Gb/s, a simple monitoring technique is required. Therefore, a novel waveform monitoring technique by prescaled-clock tone detection was proposed in a previous report. In this paper, detailed principle of the proposed technique was explained. The monitoring technique is based on an asynchronous beat signal generation using an elecro-absorption modulator (EAM) and is able to separately observe waveform distortion caused by accumulated chromatic dispersion (CD), polarization mode dispersion (PMD) and optical signal to-noise ratio (OSNR) degradation. The verification of concepts was performed by experiments, in which 1 GHz pre-scaled signals were employed to monitor distortion of OTDM 160 Gb/s carrier suppressed return-to-zero (CS-RZ) signals. Furthermore, applicability to Q factor estimation was verified by an experiment. In addition, an observation of 160 Gb/s signal by the proposed monitor was demonstrated over 120 minutes using an installed fiber in JGNII testbed. PMID- 19550515 TI - Nonlinear compression in a rod-type fiber for high energy ultrashort pulse generation. AB - We report the use of nonlinear compression in a very large mode-area rod-type photonic crystal fiber. This fiber allows the use of high energy pulses in the few microjoule range. We demonstrate the compression of 4 microJ, 338 fs pulses from a fiber chirped pulse amplification (FCPA) system down to 49 fs, 41 MW peak power pulses at a repetition rate of 200 kHz with an average power of 400 mW. The nonlinear compression setup is composed of a 5-cm-long rod-type fiber and a pair of SF10 prisms. The system was optimized to obtain good temporal quality, with a temporal Strehl ration of 86 % for the compressed 49 fs pulses. PMID- 19550516 TI - Discrimination of collagen in normal and pathological skin dermis through second order susceptibility microscopy. AB - Polarization-resolved, second harmonic generation (P-SHG) microscopy at single pixel resolution is utilized for medical diagnosis of pathological skin dermis. In analyzing the large area, pixel by pixel, second-order susceptibility of normal and pathological skin dermis, we found that P-SHG can be used to distinguish normal and dermal pathological conditions of keloid, morphea, and dermal elastolysis. Specifically, we found that the second order susceptibility tensor ratio of d(33)/d(31) for normal skins is 1.27+/-0.20, while the corresponding values for keloid, morphea, and dermal elastolysis are respectively 1.67+/-0.29, 1.79+/-0.30, and 1.75+/-0.31. We also found that the histograms of the d(33)/d(31) ratio for the pathological skins contain two peak values and are 1.5 times wider than that of the normal case, suggesting that the pathological dermal collagen fibers tend to be more structurally heterogeneous. Our work demonstrates that pixel-resolved, second-order susceptibility microscopy is effective for detecting heterogeneity in spatial distribution of collagen fibers and maybe used for future clinical diagnosis and in vivo studies of collagen pathological conditions. PMID- 19550517 TI - Observation of a bent crystal-lattice by x-ray interferometry. AB - he capability of operating a separate crystal x-ray interferometer over centimeter displacements has made it possible to observe minute strain fields of a bent crystal at the atomic scale resolution by means of phase-contrast x-ray topography. Measurement and predictive capabilities of lattice strain are key ingredients of a highly accurate measurement of the Si lattice parameter and of a determination of the number of atoms in a realization of the mass unit based on an atom mass. Here we show that the observed strain can be accurately predicted by a finite-element analysis of the crystal deformation. PMID- 19550518 TI - A single-mode laser based on asymmetric Bragg reflection waveguides. AB - A single oscillation-mode laser employing the asymmetric waveguide structure is designed and analyzed. The mode selection mechanism is realized by using an asymmetric Bragg reflection waveguide (BRW) and shown to be effective to achieve high side-mode suppression ratio (SMSR). As an example, a silicon-based quasi-one dimensional BRW with Er-doped Si-nanocrystal in the silicon oxide core is considered and illustrated for the laser structure. Guidance properties and threshold conditions are examined to verify the design procedure and performance feasibility for the single oscillation mode laser. PMID- 19550519 TI - Phase retrieval from one single phase contrast x-ray image. AB - Phase retrieval is required for achieving artifact-free x-ray phase-sensitive 3D imaging. A phase-retrieval approach based on the phase-attenuation duality with high energy x-rays can greatly facilitate for phase sensitive imaging by allowing phase retrieval from only one single projection image. The previously derived phase retrieval formula is valid only for small Fresnel propagator phases corresponding to common clinical imaging tasks. In this work we presented a new duality-based phase retrieval formula that can be applied for cases with large Fresnel propagator phases corresponding to high spatial resolution imaging. The computer simulation demonstrated superiority of this new formula over the previous phase retrieval formula in reconstructing the high frequency components of imaged objects. A modified Tikhonov regularization technique has been devised for phase retrieval in cases of very high resolution and large object-detector distance such that some Fresnel propagator phases may be close or greater than pi. This new phase retrieval formula lays the foundation for implementing high resolution phase-sensitive 3D imaging of soft tissue objects. PMID- 19550520 TI - Generation of squeezed pulses with a Sagnac loop fiber interferometer using a non soliton femtosecond laser pulse at 800 nm. AB - We experimentally demonstrate generation of a squeezed vacuum at 800 nm with a Sagnac loop fiber interferometer. When negative dispersion is properly added to an input laser pulse to compensate for the fiber dispersion, the level of squeezing is improved. A squeezed vacuum of 0.45 dB is obtained at a dispersion of -0.0157 ps(2) for the 1.5 m-long fiber loop. Since the squeezed vacuum is degraded by guided acoustic-wave Brillouin scattering (GAWBS), the noise level of the squeezing is improved by -0.3 dB at a liquid nitrogen temperature. We also demonstrate generation of photon number squeezing at -1.3 dB. PMID- 19550521 TI - Computational investigation of nanohole array based SPR sensing using phase shift. AB - We present a new high spatial resolution sensor for monitoring refractive index variations caused by binding of organic and biological molecules to the metallic surface containing arrays of nanoholes. Signal transduction is provided through detecting the optical phase change in the extraordinary optical transmission (EOT) effected by surface plasmon resonance (SPR). These 2D nanoholes are well suited for the sensor chip format in which high dense integration is readily achievable. While the sensor operates at normal illumination, practical implementation of the sensor is much easier in comparison to the traditional Kretschmann arrangement for SPR sensing. Various design parameters of the device have been studied by simulation. Our results indicate that the scheme has a shot noise limited sensitivity threshold of 4.37 x 10(-9) refractive index units (RIU) and a dynamic range of 0.17 RIU, which compare favorably with typical SPR sensors, particularly in terms of achieving high resolution and wide dynamic range sensor attributes. The phase change is also quite linear over the entire refractive index detection range. PMID- 19550522 TI - Analytic scaling analysis of high harmonic generation conversion efficiency. AB - Closed form expressions for the high harmonic generation (HHG) conversion efficiency are obtained for the plateau and cutoff regions. The presented formulas eliminate most of the computational complexity related to HHG simulations, and enable a detailed scaling analysis of HHG efficiency as a function of drive laser parameters and material properties. Moreover, in the total absence of any fitting procedure, the results show excellent agreement with experimental data reported in the literature. Thus, this paper opens new pathways for the global optimization problem of extreme ultraviolet (EUV) sources based on HHG. PMID- 19550523 TI - Polyaniline/polystyrene single-nanowire devices for highly selective optical detection of gas mixtures. AB - By using spectral analysis in the visible/near-infrared region, we demonstrate a single waveguiding polyaniline/polystyrene nanowire for highly selective detection of gas mixtures with a NH(3) detection limit on parts-per-million level and relative humidity sensing ranging from 37% to 84%. The compact and flexible sensing scheme shown here may be attractive for highly selective optical detection in complex chemical or biological environments with a single nanowire. PMID- 19550524 TI - Multifocal multiphoton excitation and time correlated single photon counting detection for 3-D fluorescence lifetime imaging. AB - We report a multifocal multiphoton time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) microscope system that uses a 16 channel multi-anode PMT detector. Multiphoton excitation minimizes out-of-focus photobleaching, multifocal excitation reduces non-linear in-plane photobleaching effects and TCSPC electronics provide photon-efficient detection of the fluorescence decay profile. TCSPC detection is less prone to bleaching- and movement-induced artefacts compared to wide-field time-gated or frequency-domain FLIM. This microscope is therefore capable of acquiring 3-D FLIM images at significantly increased speeds compared to single beam multiphoton microscopy and we demonstrate this with live cells expressing a GFP tagged protein. We also apply this system to time-lapse FLIM of NAD(P)H autofluorescence in single live cells and report measurements on the change in the fluorescence decay profile following the application of a known metabolic inhibitor. PMID- 19550525 TI - Investigating functionalized active coated nanoparticles for use in nano-sensing applications. AB - In this paper we investigate the use of active coated nanoparticles (CNPs) for nano-sensing applications. Simulation results of the optical properties of an active CNP with a 24nm radius active silica core and 6nm thick plasmonic shell made of silver that has been functionalized by an additional spherical outer layer of varying thickness and refractive index are presented. In particular, the effects of the functional-layer thickness and refractive index on the super resonant (SR) state of the active CNP are presented. It is shown that the wavelength and optical gain required to excite the SR state may provide both a spectral and a power signature usable for nano-scale sensing and that these signatures may be used to identify the dimensions and optical properties of the functional layer. These results are then applied to the case of a functional layer containing a solution of human hemoglobin. It is demonstrated that the concentration of hemoglobin may be remotely determined from these SR signatures. PMID- 19550526 TI - Development of a synchronous fluorescence imaging system and data analysis methods. AB - Although conventional autofluorescence spectroscopy, in which fluorescence emission spectra are recorded for fixed excitation wavelengths, has demonstrated good performance in tissue diagnosis, it suffers from prolonged data acquisition time and broad-band fluorescence features. Synchronous spectroscopy has been proposed to overcome the limitations of conventional fluorescence spectroscopy but has not been applied to imaging for tissue diagnosis in vivo. Our group has developed a synchronous fluorescence imaging system to combine the great diagnostic potential of synchronous spectroscopy and the large field of view of imaging for cancer diagnosis. This system has been tested in a mouse skin model to capture synchronous fluorescence images. A simple discriminant analysis method and a more complicated multi-variate statistical method have been developed to generate a single diagnostic image from a large number of raw fluorescence images. Moreover, it was demonstrated that the diagnostic image generated from synchronous data is comparable to that generated from full spectral data in classification accuracy. PMID- 19550527 TI - Self-assembled hollow waveguides with hybrid metal-dielectric Bragg claddings. AB - We report on the fabrication and characterization of integrated hollow waveguides cladded by gold-terminated, omnidirectional Bragg reflectors. The hollow waveguide channels were realized by the controlled formation of straight-sided delamination buckles within a multilayer thin film stack. An optimized process produced low-defect, straight-sided buckles with base widths from 10 to 80 mum, and corresponding peak core heights from ~0.7 to ~4 mum, on a single sample. The waveguides described have upper and lower cladding mirrors of 4 and 5.5 periods, respectively. Gold termination of the cladding reflectors significantly reduces the propagation loss of air-guided modes. The minimum propagation loss is less than 4 dB/cm in the near infrared, corresponding to upper and lower cladding reflectance of ~ 0.999. The main details of the guiding mechanism are well approximated by a simple ray-optics model. PMID- 19550528 TI - A new conformal radiation boundary condition for high accuracy finite difference analysis of open waveguides. AB - A highly accurate radiation boundary condition for finite difference analysis of open waveguides is introduced. The boundary condition is applicable to the structures embedded in a homogeneous medium and fitted to the cross section of the structure. The numerical tests carried out for a few types of waveguides including microstructured fibers showed that the proposed approach improves the accuracy by about an order of magnitude in comparison with the PML technique and eliminates all its disadvantages. PMID- 19550529 TI - The resolution of optical traps created by Light Induced Dielectrophoresis (LIDEP). AB - Light induced dielectrophoresis (LIDEP) is a variant of the dielectrophoresis (DEP) mechanism that has been used for some time to manipulate particles in a microfluidic environment. Rather than relying on lithographically created contacts to generate the required electrical fields, the electrical contacts in LIDEP are created through the selective illumination of a photoconductor. The key question we address is how microscopic traps created via LIDEP compare to optical traps based on the gradient force, in terms of power required and trap stiffness achieved, as well as the size resolution of such a trap. We highlight the complex interplay between optical power and resolution with electrical parameters, such as the electrical resistance and applied AC Voltage. We show that for a spotsize of five micrometres and larger, particles can indeed be trapped with low power. We use trap stiffness per mW to compare LIDEP with an optical trap and show that our system is 470+/- 94 times stiffer per mW than a conventional optical trap, with no loss of resolution. We also discuss the difficulties of achieving trapping at smaller spot sizes, and that the submicron resolution possible with gradient force trapping is very difficult to realise with LIDEP. PMID- 19550530 TI - An optical solution for the traveling salesman problem: erratum. AB - The initialization time for setting up the experiment grows exponential with the number of cities. Therefore the overall time required to solve the problem is not quadratic but exponential. PMID- 19550531 TI - Fabrication of photonic devices in nanostructured glasses by femtosecond laser pulses. AB - Focused femtosecond laser pulses have been used to modify the optical properties of glass doped with CdSxSe1-x nanocrystals. Large positive refractive index changes have been observed and exploited for the fabrication of photonic devices. In particular, we report on highly confining optical waveguides and single and multi-layer volume diffraction gratings. PMID- 19550532 TI - Mobility and transverse flow visualization using phase variance contrast with spectral domain optical coherence tomography. AB - Phase variance-based motion contrast is demonstrated using two phase analysis methods in a spectral domain optical coherence tomography system. Mobility contrast is demonstrated for an intensity matched Intralipid solution placed without flow within agarose wells. Vasculature oriented transversely to the imaging direction has been imaged for 3-4 dpf in vivo zebrafish using the phase variance contrast methods. 2D phase variance contrast images are demonstrated with imaging times only 25% higher than a Doppler flow image with comparable statistics. En face images created by integrating depth regions of 3D zebrafish intensity and phase variance contrast data demonstrate vasculature consistent with expected images. PMID- 19550533 TI - Effect of optical correction and remaining aberrations on peripheral resolution acuity in the human eye. AB - Retinal sampling poses a fundamental limit to resolution acuity in the periphery. However, reduced image quality from optical aberrations may also influence peripheral resolution. In this study, we investigate the impact of different degrees of optical correction on acuity in the periphery. We used an adaptive optics system to measure and modify the off-axis aberrations of the right eye of six normal subjects at 20 degrees eccentricity. The system consists of a Hartmann Shack sensor, a deformable mirror, and a channel for visual testing. Four different optical corrections were tested, ranging from foveal sphero-cylindrical correction to full correction of eccentric low- and high-order monochromatic aberrations. High-contrast visual acuity was measured in green light using a forced choice procedure with Landolt C's, viewed via the deformable mirror through a 4.8-mm artificial pupil. The Zernike terms mainly induced by eccentricity were defocus and with- and against-the-rule astigmatism and each correction condition was successfully implemented. On average, resolution decimal visual acuity improved from 0.057 to 0.061 as the total root-mean-square wavefront error changed from 1.01 mum to 0.05 mum. However, this small tendency of improvement in visual acuity with correction was not significant. The results suggest that for our experimental conditions and subjects, the resolution acuity in the periphery cannot be improved with optical correction. PMID- 19550534 TI - Proposal of three-dimensional phase contrast holographic microscopy. AB - We propose a three-dimensional phase contrast digital holographic microscopy. The object to be observed is a low-contrast transparent refractive index distribution sample, such as biological tissue. Low contrast phase objects are converted to high contrast images through the microscopy we propose. In order to gain high three-dimensional resolution, the direction of pump plane wave is scanned, and separate holographic images produced at each angle are acquired and decoded into complex amplitude in Fourier space. The three-dimensional image is reconstructed in a computer from all information acquired through the system. The resolution in the direction of the optical axis is increased by utilizing a 4pi configuration of objective lenzes. PMID- 19550535 TI - Models for guidance in kagome-structured hollow-core photonic crystal fibres. AB - We demonstrate by numerical simulation that the general features of the loss spectrum of photonic crystal fibres (PCF) with a kagome structure can be explained by simple models consisting of thin concentric hexagons or rings of glass in air. These easily analysed models provide increased understanding of the mechanism of guidance in kagome PCF, and suggest ways in which the high-loss resonances in the loss spectrum may be shifted. PMID- 19550536 TI - Multilayer 3-D photonics in silicon. AB - Three-dimensionally (3-D) integrated photonic structures in multiple layers of silicon are reported. Implantation of oxygen ions into a silicon-on-insulator substrate with a patterned thermal oxide mask, followed by a high temperature anneal, creates photonic structures on 3-D integrated layers of silicon. This process is combined with epitaxial growth to achieve devices on three vertically integrated layers of silicon. As a demonstration vehicle, we report a multistage optical filter that comprises of coupled microdisks on two subsurface silicon layers with bus waveguides on the surface (3rd) layer. The optical filter shows extinction ratios in excess of 14 dB, with excess insertion loss of less than 1 dB. PMID- 19550537 TI - Generating a geometric mode for clarifying differences between an operator method and SU(2) wave representation. AB - We study both numerically and experimentally a specific geometric mode, named VW mode, in an end-pumped Nd:YVO4 laser with a plano-concave cavity near the 1/3 degeneracy. Three theoretical methods are used to analyze the transverse profiles, the operator method [J. Opt. Soc. Am. A 22, 1559 (2005)], the SU(2) wave representation [Phys. Rev. A 69, 053807 (2004)], and the Fox-Li approach. Some differences among them are addressed and clarified. Moreover, the generating conditions for VW mode are found peculiar and its propagation character is demonstrated. By comparing the experimental mode patterns with the three numerical results, we conclude that the field of a geometric mode in the operator method should be extended to include those of the reverse directional trajectories and the SU(2) coherent state representation is found too specific to produce the fringes within some transverse patterns. PMID- 19550538 TI - Enhanced output power for phase-matched second-harmonic generation at 10.6 mum in a ZnGeP(2) crystal. AB - Second-harmonic generation was phase-matched at the fundamental wavelength of 10.6 mum in an annealed ZnGeP(2) crystal at room temperature. Our results demonstrated that the phase-matching angle was decreased with increasing the pump power. Such a unique dependence resulted in a significant enhancement on the second-harmonic output power. The highest average output power at 5.3 mum was 55 mW for an average pump power of 5.0 W, which corresponded to a conversion efficiency of 1.1%. Due to the laser-induced heating effect, the second-harmonic output power was increased by 65%. Such an efficient conversion was made possible also by using a short-pulse repetition-frequency-excited waveguide and single longitudinal-mode CO(2) laser as a fundamental beam. PMID- 19550539 TI - NIR Raman spectroscopic investigation of single mitochondria trapped by optical tweezers. AB - Raman spectroscopy is a vibration spectroscopic technique that has been widely used to probe biochemical changes of biological sample such as tumor tissue, blood cells, bacteria and yeast. Here, we applied near-infrared Raman spectroscopy to analyze the chemical composition changes of intact or swollen mitochondria induced by calcium ions. We used a confocal Laser Tweezers Raman Spectroscopy (LTRS) system that combined optical trapping and near infrared Raman spectroscopy to confine a single mitochondrion and consequently measure its Raman spectra following the addition of calcium ion solution. We analyzed Raman spectra of mitochondria isolated from rat liver, heart muscle and kidney, respectively. The major Raman peaks at 1654, 1602, 1446, 1301 and 1226 cm(-1) were observed from individual intact mitochondria. We examined the differences in near infrared spectra between intact and Ca(2+) damaged mitochondria. We found that after the exposure of the intact mitochondria to the 100 muM Ca(2+) solution the band of 1602 cm(-1) decreased very rapidly in the first period and then disappeared after 30minutes, while the intensities of the phospholipids and protein bands changed slowly in the first period and then suddenly disappeared, corresponding to the Ca(2+) induced swelling process. These results demonstrate the potential of LTRS technique as a valuable tool for the study of bioactivity and molecular composition of mitochondria. PMID- 19550540 TI - Improvement of cylindrical cloaking with the SHS lining. AB - We analyze the effectiveness of cloaking an infinite cylinder from observations by electromagnetic waves in three dimensions. We show that, as truncated approximations of the ideal permittivity and permeability material parameters tend towards the singular ideal cloaking values, the D and B fields blow up near the cloaking surface. Since the metamaterials used to implement cloaking are based on effective medium theory, the resulting large variation in D and B poses a challenge to the suitability of the field-averaged characterization of epsilon and mu. We also consider cloaking with and without the SHS (soft-and-hard surface) lining. We demonstrate numerically that cloaking is significantly improved by the SHS lining, with both the far field of the scattered wave significantly reduced and the blow up of D and B prevented. PMID- 19550541 TI - Opto-fluidic ring resonator lasers based on highly efficient resonant energy transfer. AB - We demonstrate an opto-fluidic ring resonator dye laser using highly efficient energy transfer. The active lasing material consists of a donor and acceptor mixture and flows in a fused silica capillary whose circular cross section forms a ring resonator and supports the whispering gallery modes (WGMs) of high Q factors (>107). The excited states are created in the donor and transferred to the acceptor through the fluorescence resonant energy transfer (FRET), whose emission is coupled into the WGM. Due to the high energy transfer efficiency and high Q-factors, the acceptor exhibits a lasing threshold as low as 0.3 muJ/mm2. We further analyze the energy transfer mechanisms and find that non-radiative Forster transfer is the dominant effect to support the acceptor lasing. FRET lasers using cascade energy transfer and using quantum dots (QDs) as the donor are also presented. Our study will not only lead to development of novel microfluidic lasers with low lasing thresholds and excitation/emission flexibility, but also open an avenue for future laser intra-cavity bio/chemical sensing. PMID- 19550542 TI - Selective metallization on insulator surfaces with femtosecond laser pulses. AB - We report selective metallization on surfaces of insulators (glass slides and lithium niobate crystal) based on femtosecond laser modification combined with electroless plating. The process is mainly composed of four steps: (1) formation of silver nitrate thin films on the surfaces of glass or crystal substrates; (2) generation of silver particles in the irradiated area by femtosecond laser direct writing; (3) removal of unirradiated silver nitrate films; and (4) selective electroless plating in the modified area. We discuss the mechanism of selective metallization on the insulators. Moreover, we investigate the electrical and adhesive properties of the copper microstructures patterned on the insulator surfaces, showing great potential of integrating electrical functions into lab-on a-chip devices. PMID- 19550543 TI - ABCD formalism and attosecond few-cycle pulse via chirp manipulation of a seeded free electron laser. AB - An ABCD formalism is identified to characterize a seeded Free Electron Laser (FEL) with three chirps: an initial frequency chirp in the seed Laser, an energy chirp in the electron bunch, and an intrinsic frequency chirp due to the FEL process. A scheme of generating attosecond few-cycle pulses is proposed by invoking an FEL seeded by high-order harmonic generation (HHG) from an infrared laser. The HHG seed has generic attosecond structure. It is possible to manipulate these three chirps to maintain the attosecond structure via post undulator chirped pulse compression. PMID- 19550544 TI - Group velocity inversion in AlGaAs nanowires. AB - We investigated the dispersion characteristics of submicron sized AlGaAs waveguides. Numerical simulations shows that the tight confinement of the optical waves in such nanowires leads to strong variations of the dispersion characteristics compared to classic, weakly guided waveguides of the same material system. We found numerically that the investigated structure has negative GVD for the TE mode provided the waveguide width is between 670 nm and 280 nm. Experimental data obtained from 300 mum - 1 mm long wires confirms the numerical results. PMID- 19550545 TI - Improved method of Fournier-Forand marine phase function parameterization. AB - Volume scattering functions (VSFs) and other optical seawater parameters were measured during a cruise in the Southern Baltic. Phase functions (PFs) calculated from VSFs were compared with Fournier-Forand phase functions parameterized with backscattering ratios. Due to significant divergences between experimental and modeled data a new method of Fournier-Forand phase function parameterization is proposed. PMID- 19550546 TI - Bright narrowband source of photon pairs at optical telecommunication wavelengths using a type-II periodically poled lithium niobate waveguide. AB - We report on the generation of narrowband photon pairs at telecommunication wavelengths using a periodically poled lithium niobate waveguide that utilizes the nonlinear tensor element d(24) for type-II quasi phase matching. The FWHM bandwidth of the spontaneous parametric downconversion was 1 nm. The brightness of the photon pair source was ~6x10(5)/s/GHz when the pump power was 1 mW. The indistinguishability of the signal and idler photons generated by the degenerate spontaneous parametric downconversion process was studied in a Hong-Ou-Mandel type interference experiment. PMID- 19550547 TI - Near infrared photorefractive self focusing in Sn(2)P(2)S(6):Te crystals. AB - The experimental observation of photorefractive self focusing in Sn(2)P(2)S(6) : Te bulk crystals at 1.06 mum wavelength is presented. Steady state self focusing is reached as fast as 15 ms for an input peak intensity equal to 160 W/cm(2). Self focusing is maximum for input peak intensities around 15 W/cm(2) and is decreasing for intensities below and above this value. PMID- 19550548 TI - Highly efficient fluorescence sensing with hollow core photonic crystal fibers. AB - We investigate the potential of microstructured optical fibers (MOFs) for highly sensitive absorption and fluorescence measurements by infiltrating a dye solution in the holey structure. Generally in a MOF only the evanescent part of the electromagnetic field penetrates into the sample material, providing a weak light matter interaction. We compare such a MOF with a selectively filled hollow core photonic crystal fiber (HCPCF), in which most of the field energy propagates in the sample material. We show that dye concentrations down to 1x10(-10) M can be detected in a HCPCF using only nanoliter sample volumes. Our experiments proof that HCPCFs are well suited for demanding sensing applications, outperforming existing fiber tools that rely on evanescent sensing. PMID- 19550549 TI - Fourier transform channeled spectropolarimetry in the MWIR. AB - A complete Fourier Transform Spectropolarimeter in the MWIR is demonstrated. The channeled spectral technique, originally developed by K. Oka, is implemented with the use of two Yttrium Vanadate (YVO(4)) crystal retarders. A basic mathematical model for the system is presented, showing that all the Stokes parameters are directly present in the interferogram. Theoretical results are compared with real data from the system, an improved model is provided to simulate the effects of absorption within the crystal, and a modified calibration technique is introduced to account for this absorption. Lastly, effects due to interferometer instabilities on the reconstructions, including nonuniform sampling and interferograms translations, are investigated and techniques are employed to mitigate them. PMID- 19550550 TI - Near-field coupling of a single fluorescent molecule and a spherical gold nanoparticle. AB - Near-field coupling of a single gold nanoparticle (GNP) to a single fluorescent molecule is investigated here for varying separation d between the two. While the emission quantum efficiency of the coupled system generally decreases for d!0, a pronounced near-field enhancement is observed under certain conditions, partly outweighing the efficiency loss at small distances. We report on optimizing these conditions by varying the excitation field direction and the three-dimensional relative configuration between the GNP and the fluorophore. Furthermore, we examine how the sphere diameter, the surrounding medium, as well as the absorption and emission wavelengths of the molecular dipole influence the fluorescence yield. Our results are of high practical relevance for all GNP mediated application fields such as fluorescence microscopy, scattering near field optical microscopy, bioanalytics, and medical applications. PMID- 19550551 TI - Multi-photon excitation properties of CdSe quantum dots solutions and optical limiting behavior in infrared range. AB - Multi-photon absorption and excitation properties of CdSe quantum dots in hexane with different dot-sizes have been investigated. The two- and three-photon absorption (2PA and 3PA) coefficients were measured by using ~160-fs laser pulses at wavelengths of ~775-nm and ~1300-nm, respectively. The dependence of one-, two and three-photon induced fluorescence spectra as well as their double exponential decay on the dot-sizes was studied. Based on the fluorescence emission spectra and temporal decay constants for a given sample solution excited by one-, two-and three-photon absorption, it can be concluded that the transition pathways for fluorescence emission and decay under one-, two- and three-photon excitation are nearly identical. The optical power limiting capabilities based on 2PA and 3PA mechanisms are demonstrated separately. In addition, a saturation behavior of 3PA at ~1300 nm was observed. PMID- 19550552 TI - Near-infrared light scattering by particles in coastal waters. AB - We report the first measurements of the scattering coefficient of natural marine particles, which extend over the near-infrared spectral region to up to 870 nm. The measurements were conducted in three different European estuaries (Gironde, Tamar and Elbe) using an in situ absorption and attenuation-meter. The observed particulate scattering coefficients varied from 1 to nearly 100 m(-1). The spectral shape in the near-infrared very closely matched a lambda(-gamma) spectral dependence, which is expected when the particle size followed a power law distribution. The spectral slope of the scattering spectrum, gamma, spanned from 0.1 to 1.2 and showed significant regional and temporal variations. These variations were certainly related to the particle size distribution, which will have to be studied in future works. Using our near-infrared data as a reference, we assessed the use of the attenuation coefficient spectrum in the visible range to estimate the near-infrared particulate scattering slope and found values different by 10% on average. PMID- 19550553 TI - Fresnel rhombs as achromatic phase shifters for infrared nulling interferometry. AB - We propose a new family of achromatic phase shifters for infrared nulling interferometry. These key optical components can be seen as optimized Fresnel rhombs, using the total internal reflection phenomenon, modulated or not. The total internal reflection indeed comes with a phase shift between the polarization components of the incident light. We propose a solution to implement this vectorial phase shift between interferometer arms to provide the destructive interference process needed to disentangle highly contrasted objects from one another. We also show that, modulating the index transition at the total internal reflection interface allows compensating for the intrinsic material dispersion in order to make the subsequent phase shift achromatic over especially broad bands. The modulation can be induced by a thin film of a well-chosen material or a subwavelength grating whose structural parameters are thoroughly optimized. We present results from theoretical simulations together with preliminary fabrication outcomes and measurements for a prototype in Zinc Selenide. PMID- 19550554 TI - Bending sensitivity of long-period fiber gratings inscribed in holey fibers depending on an axial rotation angle. AB - We discuss bending properties of a long-period fiber grating (LPFG) inscribed into a holey fiber (HF) depending on an axial rotation angle. High quality of the HF-based LPFG with a high extinction ratio of more than 20 dB is achieved. The proposed HF-based LPFG has bending insensitivity under a certain range of the bending curvature. As the bending curvature is higher than 4 m(-1), the center wavelength of the grating is shifted into the shorter wavelength. Bending sensitivity of the HF-based LPFG is changed by an axial rotation angle, which shows its dependence on the rotational orientation. We measure the transmission characteristics of the HF-based LPFG with the ambient index change. The HF-based LPFG has ambient index insensitivity because of the air holes in the inner cladding. PMID- 19550555 TI - Crossing and branching nodes in soft-lithography-based optical interconnects. AB - We investigate the characteristics of crossing and branching nodes in monolayer soft-lithography-based polymer optical interconnects with experimental and theoretical analysis. The theoretical crosstalk, as calculated by a function of crossing angle, was determined for a set of interconnect pairs with varying cross sections, and was compared with experimental measurements. Furthermore, a suitable branching angle was found for branching node and the effects of short distance mode scrambling in highly multimode polymer waveguides were studied in detail. It was found that mode-filling occurred within a propagation distance of 1.5mm for a 50x50mum2 cross-section for VCSEL coupling; however, complete scrambling of ray direction required a propagation distance of more than 5mm. PMID- 19550556 TI - Frequency-doubling of a CW fiber laser using PPKTP, PPMgSLT, and PPMgLN. AB - Second-harmonic generation (SHG) in PPKTP, PPMgSLT, and PPMgLN crystals is analyzed by frequency-doubling CW light from a 1064 nm fiber laser over a range of powers up to 10 W. Data for fundamental powers less than 3 W is used to determine the effects of the fundamental laser linewidth on SHG and to identify imperfections in the periodicity and boundary sharpness of the crystals' poled domains which can reduce SHG. Data for fundamental powers greater than 3 W is used to diagnose and model limiting effects on SHG such as pump depletion and thermal dephasing. Thermal dephasing was found to reduce second-harmonic power by 25% or more for input fundamental powers approaching 10 W. PMID- 19550557 TI - Experimental study on measurement of aspheric surface shape with complementary annular subaperture interferometric method. AB - Based on our previously reported annular subaperture reconstruction algorithm with Zernike annular polynomials and matrix method, we provide an experimental demonstration by testing a parabolic mirror with the complementary annular subaperture interferometric method. By comparing the results of annular subaperture method with that of the classical auto-collimation method, it is shown that the reconstruction results are in good agreement with the auto collimation measurement. In addition, we discuss some limitations of characterizing annular subaperture measurement data with finite Zernike coefficients in our algorithm, and also show the possibility of characterizing higher spatial frequency information with adequate Zernike coefficients. It is believable that the reported method can be extended to test the surface shape of some large concave aspheric mirrors with acceptable accuracy. PMID- 19550558 TI - Continuous-wave and Q-switched laser operation of Yb:NaY(WO(4))(2) crystal. AB - Diode-pumped continuous-wave (cw) and Q-switched laser operations were demonstrated, for the first time to our knowledge, with a disordered Yb:NaY(WO(4))(2) crystal. A cw output power of 5.2 W at 1045 nm was generated with an optical-to-optical efficiency of 37%. The slope efficiency in the absence of thermal losses was 50%. With a Cr(4+):YAG saturable absorber to passively Q switch the laser, an average output power of 2.0 W was obtained with a slope efficiency of 35%. The output pulse energy, duration, and peak power were 145 muJ, 26 ns, and 5.6 kW, respectively. PMID- 19550560 TI - Switching with vortex beams in nonlinear concentric couplers. AB - We demonstrate that a concentric ring coupler can be employed for nonlinear switching of the angular momentum of light carried by an optical vortex. We find different types of stationary vortex states in the nonlinear coupler and study coupling of both power and momentum of an optical vortex launched into one of the rings, demonstrating that the switching takes place well below the collapse threshold. The switching is more effective for the inner-ring excitation since it triggers more sharply and for the powers low enough to avoid the vortex instability and breakup. PMID- 19550559 TI - Enhancement of Nd:YAG LIBS emission of a remote target using a simultaneous CO(2) laser pulse. AB - For the first time to the best of our knowledge, a simultaneous 10.6 mum CO(2) laser pulse has been used to enhance the Laser Induced Breakdown Spectroscopy (LIBS) emission from a 1.064 mum Nd:YAG laser induced plasma on a hard target. The enhancement factor was on the order of 25 to 300 times, depending upon the emission lines observed. For an alumina ceramic substrate the Al emission lines at 308 nm and Fe impurity line at 278 nm showed an increase of 60x and 119x, respectively. The output energy of the Nd:YAG laser was 50 mJ/pulse focused to a 1 mm diameter spot to produce breakdown. The CO(2) laser pulse had a similar energy density of 40 mJ/mm(2). Timing overlap of the two laser pulses within 1 microsecond was important for enhancement to be observed. An observed feature was the differential enhancement between different elemental species and also between different ionization states, which may be helpful in the application of LIBS for multi-element analysis. PMID- 19550561 TI - Geometric distortion-invariant watermarking based on Tschebycheff transform and dual-channel detection. AB - Many proposed image watermarking techniques are sensitive to geometric distortions such as rotation, scaling, and translation. Geometric distortion, even by a slight amount, can disable a watermark decoder. In this study, a geometric distortion-invariant watermarking technique is designed by utilizing Tschebycheff moments of the original image to estimate the geometric distortion parameters of corrupted watermarked images. The Tschebycheff moments of an original image can be used as a private key for watermark extraction. The embedding process is a closed-loop system that modifies the embedding intensity according to the results of the performance analysis. The convergence of the closed-loop system is proved. Different from early heuristic methods, the optimal blind watermark detector is designed with the introduction of dual-channel detection utilizing high-order spectra detection and likelihood detection. Even with a small signal-to-noise ratio (SNR), the detector can still get a satisfying detection probability if there is enough high-order spectra information. When the high-order spectra are small, this dual-channel detection system will become a likelihood detection system. The watermark decoder extracts a watermark by blindly utilizing independent component analysis (ICA). The computational aspects of the proposed watermarking technique are also discussed in detail. Experimental results demonstrate that the proposed watermarking technique is robust with respect to attacks performed by the popular watermark benchmark, StirMark. PMID- 19550562 TI - Bistability and all-optical switching in semiconductor ring lasers. AB - Semiconductor ring lasers display a variety of dynamical regimes originating from the nonlinear competition between the clockwise and counter-clockwise propagating modes. In particular, for large pumping the system has a bistable regime in which two stationary quasi-unidirectional counter-propagating modes coexist. Bistability is induced by cross-gain saturation of the two counter-propagating modes being stronger than the self-saturation and can be used for data storage when the semiconductor ring laser is addressed with an optical pulse. In this work we study the response time when an optical pulse is injected in order to make the system switch from one mode to the counter-propagating one. We also determine the optimal pulse energy to induce switching. PMID- 19550563 TI - Broad-band continuous-wave parametric wavelength conversion in silicon nanowaveguides. AB - We demonstrate highly broad-band frequency conversion via four-wave mixing in silicon nanowaveguides. Through appropriate engineering of the waveguide dimensions, conversion bandwidths greater than 150 nm are achieved and peak conversion efficiencies of -9.6 dB are demonstrated. Furthermore, utilizing fourth-order dispersion, wave-length conversion across four telecommunication bands from 1477 nm (S-band) to 1672 nm (U-band) is demonstrated with an efficiency of -12 dB. PMID- 19550564 TI - Analysis of whispering-gallery microcavity-enhanced chemical absorption sensors. AB - A theoretical analysis of the operation of a chemical sensor based on cavity enhanced optical absorption is given for a system in which the cavity is a dielectric whispering-gallery microresonator. Continuous-wave input is assumed, and the detection sensitivity is characterized in terms of an effective absorption path length. In the case of tunable single-frequency input, it is shown that monitoring analyte-induced changes in the throughput dip depth enables detection with relative sensitivity greater than that of frequency-shift and cavity-ringdown methods. In addition, for the case of broadband input and drop port output, an analysis applicable to microcavity-enhanced absorbance spectroscopy experiments is given. PMID- 19550565 TI - Fiber nonlinearity pre- and post-compensation for long-haul optical links using OFDM. AB - The nonlinear power limit of optical links using optical Orthogonal Frequency Division Multiplexing (OFDM) for dispersion compensation can be significantly improved using an optimum combination of nonlinearity precompensation and postcompensation. The compensation is implemented at the transmitter and at the receiver as computationally-efficient power-dependent phase shifts with a single tuning parameter. The system is robust against the exact details of the fiber plant's dispersion and power levels. Using an optimum combination of pre and post compensation allows a 2-dB increase in launch power for 2000-km standard single mode fiber (S-SMF) systems and 5-dB when 6 ps/nm/km fibers are used. Using pre or post compensation alone approximately halves these values. PMID- 19550566 TI - Thermally induced wavelength tunability of microcavity solid-state dye lasers. AB - Wavelength tunability of a microcavity solid-state dye laser is modeled and demonstrated by simulations making use of the finite element method. We investigate the application of two phenomena, thermoelastic expansion of the microcavity material and thermo-induced change of the refractive index, to tune the microcavity mode frequencies by a variation of the effective optical path. An optimized size of the laser microcavity is defined depending on the operation wavelength bandwidth and the glass transition temperature of the gain material. PMID- 19550567 TI - Inverse silica opal photonic crystals for optical sensing applications. AB - This work reports fabrication of inverse silica opal photonic crystal structures from direct polystyrene micro sphere opals using low-temperature sol-gel infiltration of silica, and examines performance of these photonic crystals as environmental refractive index sensors. Sensitivity of the spectral position and optical attenuation of photonic stop gaps is found to allow detection of the index changes by the amount of ~10(-3). The high value of sensitivity, which is comparable with those of other optical sensing techniques, along with simplicity of the optical detection setup required for sensing, and the low-temperature, energy-efficient fabrication process make inverse silica opals attractive systems for optical sensing applications. PMID- 19550568 TI - Waveplate analyzer using binary magneto-optic rotators. AB - We demonstrate a simple waveplate analyzer to characterize linear retarders using magneto-optic (MO) polarization rotators. The all-solid state device can provide highly accurate measurements for both the retardation of the waveplate and the orientation of optical axes simultaneously. PMID- 19550569 TI - Plasmonic Bragg reflectors for enhanced extraordinary optical transmission through nano-hole arrays in a gold film. AB - We demonstrate the use of plasmonic Bragg reflectors (PBRs) to enhance the extraordinary optical transmission (EOT) from an array of subwavelength apertures in a gold film. Arrays of partially milled lines and dimples are placed at the edges of an array of nano-holes in a gold film. These PBR structures, with half the pitch of the array, capture light scattered away from the array by Bragg reflection. By appropriate positioning of the PBR, the light is reflected in phase with the EOT light and thereby doubles the EOT without shifting the wavelength of the resonant transmission peak. Furthermore, the PBR structures show strong polarization dependence that is also strongly dependent on the structure of the PBR, as explained in terms of scattering of the surface waves. PMID- 19550570 TI - Investigation of the suitability of silicate bonding for facet termination in active fiber devices. AB - We demonstrate that silicate bonding an optical flat to the output facet of an active fiber device can increase the reliability of high-peak power systems and subsantially reduce the effective feedback at the termination of a double-clad fiber. We determine the bonding parameters and conditions that maximize the optical damage threshold of the bond and minimize the Fresnel reflection from the bond. At 1-mum wavelength, damage thresholds greater than 70 J/cm(2) are demonstrated for 25-ns pulses. We also measured Fresnel reflections less than -63 dB off the bond. Finally, we determined that the strength of the bond is sufficient for most operating environments. PMID- 19550571 TI - Viewing region maximization of an integral floating display through location adjustment of viewing window. AB - Integral floating display is a recently proposed three-dimensional (3D) display method which provides a dynamic 3D image in the vicinity to an observer. It has a viewing window only through which correct 3D images can be observed. However, the positional difference between the viewing window and the floating image causes limited viewing zone in integral floating system. In this paper, we provide the principle and experimental results of the location adjustment of the viewing window of the integral floating display system by modifying the elemental image region for integral imaging. We explain the characteristics of the viewing window and propose how to move the viewing window to maximize the viewing zone. PMID- 19550572 TI - PINIP based high-speed high-extinction ratio micron-size silicon electrooptic modulator. AB - We propose an electrooptic device in silicon based on a p-i-n-i-p device structure for charge transport. The proposed device exhibits carrier injection time of 10 ps and extraction time of 15 ps enabling 100 GHz operation. When integrated into a resonator the proposed micron-size device operates at 40 Gb/s with 12 dB extinction ratio and 4fJ/bit/micron-length power dissipation, limited in speed only by the photon lifetime of the resonator. PMID- 19550573 TI - Low-threshold, room-temperature femtosecond Cr4+:forsterite laser. AB - We describe a low-threshold Cr(4+):forsterite laser which can be operated in continuous-wave as well as Kerr-lens mode-locked regimes at room temperature. To achieve low lasing thresholds, a 3-mm-long Cr(4+):forsterite crystal with a single-pass absorption of 41% was used to reduce the passive losses. Furthermore, a thin crystal was used to minimize the pump induced thermal gradients. During continuous-wave operation, absorbed threshold pump powers as low as 290 mW were obtained at 200C in single-pass pumping with a Yb fiber laser. To the best of our knowledge, this is the lowest pump threshold reported to date for Cr(4+):forsterite lasers. Furthermore, Kerr-lens mode locking could also be obtained using dispersion compensation with double-chirped mirrors. With an input pump power of 3 W, nearly transform-limited 89-fs pulses were obtained at a repetition rate of 200 MHz and with an average output power of 85 mW. PMID- 19550574 TI - Nano-lithographically fabricated titanium dioxide based visible frequency three dimensional gap photonic crystal. AB - Photonic crystals (PC) have emerged as important types of structures for light manipulation. Ultimate control of light is possible by creating PCs with a complete three dimensional (3D) gap [1, 2]. This has proven to be a considerable challenge in the visible and ultraviolet frequencies mainly due to complications in integrating transparent, high refractive index (n) materials with fabrication techniques to create ~ 100nm features with long range translational order. In this letter, we demonstrate a nano-lithography approach based on a multilevel electron beam direct write and physical vapor deposition, to fabricate four-layer titania woodpile PCs that potentially exhibit complete 3D gap at visible wavelengths. We achieved a short wavelength bandedge of 525nm with a 300nm lattice constant PC. Due to the nanoscale precision and capability for defect control, the nanolithography approach represents an important step toward novel visible photonic devices for lighting, lasers, sensing and biophotonics. PMID- 19550575 TI - Shaping and control of polychromatic light in nonlinear photonic lattices. AB -

Focus Serial: Frontiers of Nonlinear Optics

We overview our recent results on spatio-spectral control, diffraction management, broadband switching, and self-trapping of polychromatic light in periodic photonic lattices in the form of rainbow gap solitons, polychromatic surface waves, and multigap color breathers. We show that an interplay of wave scattering from a periodic structure and interaction of multiple colors in media with slow nonlinear response can be used to selectively separate or combine different spectral components. We use an array of optical waveguides fabricated in a LiNbO(3) crystal to actively control the output spectrum of the supercontinuum radiation and generate polychromatic gap solitons through a sharp transition from spatial separation of spectral components to the simultaneous spatio-spectral localization of supercontinuum light. We also show that by introducing specially optimized periodic bending of waveguides in the longitudinal direction, one can manage the strength and type of diffraction in an ultra-broad spectral region and, in particular, realize the multicolor Talbot effect. PMID- 19550576 TI - Spatio-temporal reshaping and X Wave dynamics in optical filaments. AB -

Focus Serial: Frontiers of Nonlinear Optics

We investigate ultrashort laser pulse filamentation within the framework of spontaneous X Wave formation. After a brief overview of the filamentation process we study the case of an intense filament co-propagating with a weaker seed pulse. The filament is shown to induce strong Cross-Phase Modulation (XPM) effects on the weak seed pulse: driven by the pump, the seed pulse undergoes pulse splitting with the daughter pulses slaved to their pump counterparts. They undergo strong spatio-temporal reshaping and are transformed into XWaves traveling at the same group velocities as the pump split-off pulses. In the presence of a gain mechanism such as Four-Wave-Mixing or Stimulated Raman Scattering, energy is then transferred from the pump filament leading to amplification of the seed X Wave and formation of a temporally compressed intensity peak. PMID- 19550577 TI - Experimental observations on the response of 1(st) and 2(nd) order fibre optic long period grating coupling bands to the deposition of nanostructured coatings. AB - The sensitivity of attenuation bands corresponding to the 2(nd) order coupling to cladding modes by a fibre optic long period grating (LPG) to the deposition of nanostructured coatings is investigated and compared with that of the 1(st) order coupling. The experimental observations support previously reported theoretical descriptions of LPGs with nanoscale coatings. PMID- 19550578 TI - Antireflection of transparent polymers by advanced plasma etching procedures. AB - Self-organized nanostructures that provide antireflection properties grow on PMMA caused by plasma ion etching. A new procedure uses a thin initial layer prior to the etching step. Different types of antireflective structures can now be produced in a shorter time and with fewer limitations on the type of polymer that can be used. The durability of the structured surfaces can be improved by the deposition of additional thin films. PMID- 19550579 TI - Simple nonlinear interferometer-based all-optical thresholder and its applications for optical CDMA. AB - We present an experimental demonstration of an ultrafast all-optical thresholder based on a nonlinear Sagnac interferometer. The proposed design is intended for operation at very small nonlinear phase shifts. Therefore, it requires an in-loop nonlinearity lower than for the classical nonlinear loop mirror scheme. Only 15 meters of conventional (non-holey) silica-based fiber is used as a nonlinear element. The proposed thresholder is polarization insensitive and is good for multi-wavelength operation, meeting all the requirements for autocorrelation detection in various optical CDMA communication systems. The observed cubic transfer function is superior to the quadratic transfer function of second harmonic generation-based thresholders. PMID- 19550580 TI - Broadcast MIMO over multimode optical interconnects by modal beamforming. AB - We introduce broadcast MIMO communication systems over multimode optical fibers or waveguides. Based on BeamForming (BF) at the transmitter, decoupled virtual subchannels are provided to multiple uncoordinated conventional direct detection receivers. This optical technique, extending Zero-Forcing BF wireless MIMO techniques to quadratic detection, is applicable to photonic interconnects, e.g. short-reach point-to-(multi)point transmission over MMF, up to rates of 100 Gb/s for distances up to 100 m. PMID- 19550581 TI - Dependence of extrinsic loss on group velocity in photonic crystal waveguides. AB - We examine the effects of disorder on propagation loss as a function of group velocity for W1 photonic crystal (PhC) waveguides. Disorder is deliberately and controllably introduced into the photonic crystal by pseudo-randomly displacing the holes of the photonic lattice. This allows us to clearly distinguish two types of loss. Away from the band-edge and for moderately slow light (group velocity c/20-c/30) loss scales sub-linearly with group velocity, whereas near the band-edge, reflection loss increases dramatically due to the random and local shift of the band-edge. The optical analysis also shows that the random fabrication errors of our structures, made on a standard e-beam lithography system, are below 1 nm root mean square. PMID- 19550582 TI - Femtosecond micromachining in transparent bulk materials using an anamorphic lens. AB - A unique anamorphic lens design was applied to a circular 780nm femtosecond laser pulse to transform it into an elliptically shaped beam at focus. This lens was developed to give an alternative method of micromachining bulk transparent materials. The challenge for femtosecond laser processing is to control the nonlinear affect of self-focusing, which can occur when using a fast f-number lens. Once the focused spot is dominated by self-focusing the predicted focused beam becomes a filament inside the bulk, which is an undesirable effect. The anamorphic lens resolves this self-focusing by increasing the numerical aperture (NA) and employing an elliptical beam shape. The anamorphic lens was designed to furnish a 2.5mum by 190mum line at focus. Provided the pulse energy is high enough, transparent bulk material will be damaged with a single femtosecond laser pulse. Damage in this text refers to visual change in the index of refraction as observed under an optical microscope. Using this elliptical shape (or line), grating structures were micro-machined on the surface of SiC bulk transparent substrate. SiC was chosen because it is known for its micromachining difficulty and its crystalline structure. From the lack of self-focusing and using energy that is just above the damage threshold the focused line beam generated from the anamorphic lens grating structures produced a line shape nearly identical to the geometrical approximation. In this paper we discuss a new method of writing gratings (or other types of structures) in bulk transparent materials using a single femtosecond laser pulse. We will investigate the grating structures visually (inspected under an optical microscope) and also by use of an atomic force microscopy (AFM). In addition, we test the grating diffraction efficiency (DE) as a function of grating spacing, d. PMID- 19550583 TI - On the relationship between Bloch modes and phase-related refractive index of photonic crystals. AB - It has previously been shown that the phase-related refractive index is positive in photonic crystals that display negative refraction at higher bands. We hypothesize that the phase velocity is governed by a wave that can be related to the dominant Bloch mode. This dominant wave can be identified from an approximate solution of Maxwell Equations using a homogeneously averaged dielectric constant and the dominant wavevector is related to the fundamental wavevector and the reciprocal lattice vectors. We validate this hypothesis by numerical Fourier decomposition of the field in the entire simulation domain. It confirms that for negative refraction at higher bands, the phase-related refractive index is indeed positive and differs significantly from the negative value of effective refractive index calculated from the band structure. PMID- 19550584 TI - A passively mode-locked fiber laser at 1.54 mum with a fundamental repetition frequency reaching 2 GHz. AB - We demonstrate a fundamentally mode-locked fiber laser with a repetition frequency in excess of 2 GHz at a central wavelength of 1.535 mum. Co-doped ytterbium-erbium fiber provides the gain medium for the laser, affording high gain per unit length, while a semiconductor saturable absorber mirror (SAM) provides the pulse shaping mechanism in a standing wave cavity. Results are shown confirming cw mode-locking for 1 GHz and 2 GHz repetition frequency systems. The response of the frequency comb output to pump power variations is shown to follow a single pole response. The timing jitter of a 540MHz repetition-rate laser has been suppressed to below 100 fs through phase-lead compensated feedback to the pump power. Alternatively, a single comb line of a 850MHz repetition-rate laser has been phase-locked to a narrow linewidth cw laser with an in-loop phase jitter of 0.06 rad(2). The laser design is compatible with low-noise oscillator applications. PMID- 19550585 TI - Color-encoded fringe projection for 3D shape measurements. AB - A novel technique using color-encoded stripes embedded into a sinusoidal fringe pattern for finding the absolute shape of an object is proposed. Phases of the projected fringes on the surface are evaluated by Fourier transform method. Unwrapping is then performed with reference to the color-encoded stripes. When surfaces of interest contain large depth discontinuities, the color-encoded stripes can easily identify the fringe order. Compared with other phase unwrapping schemes, this method offers many major advantages, including: (1) very low computation cost for the 3D reconstruction, (2) reliable phase unwrapping to complex objects, especially for surfaces with large depth discontinuities, (3) only one-shot measurement is required, and (4) robust performance to analyze dynamic objects. PMID- 19550586 TI - Effects of absorption on multiple scattering by random particulate media: exact results. AB - We employ the numerically exact superposition T-matrix method to perform extensive computations of elec nottromagnetic scattering by a volume of discrete random medium densely filled with increasingly absorbing as well as non-absorbing particles. Our numerical data demonstrate that increasing absorption diminishes and nearly extinguishes certain optical effects such as depolarization and coherent backscattering and increases the angular width of coherent backscattering patterns. This result corroborates the multiple-scattering origin of such effects and further demonstrates the heuristic value of the concept of multiple scattering even in application to densely packed particulate media. PMID- 19550587 TI - Electromagnetic scattering by a fixed finite object embedded in an absorbing medium. AB - This paper presents a general and systematic analysis of the problem of electromagnetic scattering by an arbitrary finite fixed object embedded in an absorbing, homogeneous, isotropic, and unbounded medium. The volume integral equation is used to derive generalized formulas of the far-field approximation. The latter serve to introduce direct optical observables such as the phase and extinction matrices. The differences between the generalized equations and their counterparts describing electromagnetic scattering by an object embedded in a non absorbing medium are discussed. PMID- 19550588 TI - Optical pulse compression in dispersion decreasing photonic crystal fiber. AB - Improvements to tapered photonic crystal fiber (PCF) fabrication have allowed us to make up to 50 m long PCF tapers with loss as low as 30 dB/km. We discuss the design constraints for tapered PCFs used for adiabatic soliton compression and demonstrate over 15 times compression of pulses from over 830 fs to 55 fs duration at a wavelength of 1.06 lm, an order of magnitude improvement over previous results. PMID- 19550589 TI - Generation of 1.5 microJ single-cycle terahertz pulses by optical rectification from a large aperture ZnTe crystal. AB - We demonstrate the generation muJ-level, single-cycle terahertz pulses by optical rectification from a large-aperture ZnTe single crystal wafer. Energies up to 1.5 muJ per pulse and a spectral range extending to 3 THz were obtained using a 100 Hz Ti:sapphire laser source and a 75-mmdiameter, 0.5-mm-thick, (110) ZnTe crystal, corresponding to an average power of 150 muW and an energy conversion efficiency of 3.1 x 10(-5). We also demonstrate real-time imaging of the focused terahertz beam using a pyroelectric infrared camera. PMID- 19550590 TI - Novel optical properties of six-fold symmetric photonic quasicrystal fibers. AB - We have investigated optical properties of an optical fiber having a six-fold symmetric quasiperiodic array of air holes in cladding, a six-fold symmetric photonic quasicrystal fiber. The photonic quasicrystal fiber exhibits larger cutoff ratio for endlessly single mode operation than that of a triangular photonic crystal fiber having six-fold symmetry and almost zero ultra-flattened chromatic dispersion, 0+/-0.05 ps/km/nm, in the range of wavelength from 1490 to 1680 nm. The dispersion value is much less than those of the proposed dispersion flattened PCFs. PMID- 19550591 TI - Near-field imaging of quantum cascade laser transverse modes. AB - We report near field imaging of the transverse lasing modes of quantum cascade lasers. A mid-infrared apertureless near-field scanning optical microscope was used to characterize the modes on the laser facet. A very stable mode pattern corresponding to a TM(00) mode was observed as function of increasing driving current for a narrow active region quantum cascade laser. Higher order modes were observed for devices with a larger active region width-to-wavelength ratio operated in pulsed mode close to threshold. A theoretical model is proposed to explain why specific transverse modes are preferred close to threshold. The model is in good agreement with the experimental results. PMID- 19550592 TI - Investigation of optical properties of circular spiral photonic crystals. AB - The photonic bandgap of three-dimensional photonic crystals, formed by arranging circular spirals in face-centre-cubic lattice, was theoretically investigated. The structure was found to have a relative photonic bandgap of up to 25% in both direct and inversed configurations. The conditions under which the structure has a bandgap larger than 10% are described. Some considerations for optimizing such photonic crystal fabrication by two-photon polymerization are given. The theoretical results are implemented to fabricate polymeric structures that can be used as templates for photonic crystals with full photonic bandgap larger than 10% centered in the near-infrared region. PMID- 19550593 TI - Improved mastering material for multilevel blue laser disc. AB - In order to get a small-sized pit, the reaction threshold of a photoresist is enhanced by increasing the prebake temperature and time. With the improved photoresist, a pit size corresponding to or even smaller than the minimum pit on a blue laser disc can be obtained on an industrial DVD product line. The improved photoresist may serve as the master material of future multilevel blue laser discs, which might provide a potential solution for the fabrication of multilevel blue laser discs. PMID- 19550594 TI - Synthesizing computer generated holograms with reduced number of perspective projections. AB - We present an improved method for recording a synthesized Fourier hologram under incoherent white-light illumination. The advantage of the method is that the number of real projections needed for generating the hologram is significantly reduced. The new method, designated as synthetic projection holography, is demonstrated experimentally. We show that the synthetic projection holography barely affects the reconstructed images. However, by increasing the number of observed projections one can improve the synthetic projection hologram quality. PMID- 19550595 TI - Optical burst add-drop multiplexing technique for sub-wavelength granularity in wavelength multiplexed ring networks. AB - We demonstrate optical burst add-drop multiplexing as a practical application of the optical burst switching technology in a wavelength-division-multiplexed ring network. To control optical bursts in the network, a burst identifier (BI) for delivering control information, and a BI processor for handling the BI, were designed. Optical bursts of 10- to 100-mus in length were optically multiplexed or demultiplexed in an intermediate node of the ring network. The demonstration shows that the optical burst add-drop multiplexing technique provides sub wavelength granularity to a ring network. PMID- 19550596 TI - Femtosecond laser written surface waveguides fabricated in Nd:YAG ceramics. AB - Near surface channel waveguides have been fabricated in Neodymium doped YAG ceramics by using IR femtosecond laser irradiation at the low frequency regime. Single mode guidance has been demonstrated with propagation losses of ~1 dB/cm. Time resolved confocal micro-luminescence experiments have been used to determine the spectroscopic properties of the Nd(3+) laser ions in the channel waveguide as well as to elucidate the waveguide formation processes. PMID- 19550597 TI - Bowtie plasmonic quantum cascade laser antenna. AB - We report a bowtie plasmonic quantum cascade laser antenna that can confine coherent mid-infrared radiation well below the diffraction limit. The antenna is fabricated on the facet of a mid-infrared quantum cascade laser and consists of a pair of gold fan-like segments, whose narrow ends are separated by a nanometric gap. Compared with a nano-rod antenna composed of a pair of nano-rods, the bowtie antenna efficiently suppresses the field enhancement at the outer ends of the structure, making it more suitable for spatially-resolved high-resolution chemical and biological imaging and spectroscopy. The antenna near field is characterized by an apertureless near-field scanning optical microscope; field confinement as small as 130 nm is demonstrated at a wavelength of 7.0 mum. PMID- 19550598 TI - Calculation of power limit due to fiber nonlinearity in optical OFDM systems. AB - We develop a simple formula for estimating the effect of Four- Wave Mixing (FWM) on received signal quality in coherent optical systems using Orthogonal Frequency Division Multiplexing (OFDM) for dispersion compensation. This shows the nonlinear limit is substantially independent of the number of OFDM subcarriers. Our analysis agrees well with full split-step Fourier method simulations, so allows the nonlinear limit of multi-span systems to be estimated without lengthy simulations. PMID- 19550599 TI - Improving the viewing angle properties of microcavity OLEDs by using dispersive gratings. AB - The changes of emission peak wavelength and angular intensity with viewing angles have been issues for the use of microcavity OLEDs. We will investigate Distributed Bragg Gratings (DBRs) constructed from largely dispersive index materials for reducing the viewing angle dependence. A DBR stack mirror, aiming at a symmetric structure and less number of grating period for a practical fabrication, is studied to achieve a chirp-featured grating for OLEDs with blue emission peak of 450nm. For maximizing the compensation of the viewing angle dependence, the effects of dispersive index, grating structure, thickness of each layer of the grating, grating period and chirp will be comprehensively investigated. The contributions of TE and TM modes to the angular emission power will be analyzed for the grating optimization, which have not been expressed in detail. In studying the light emission of OLEDs, we will investigate the Purcell effect which is important but has not been properly considered. Our results show that with a proper design of the DBR, not only a wider viewing angle can be achieved but also the color purity of OLEDs can be improved. PMID- 19550600 TI - Laser noise compression by filamentation at 400 nm in argon. AB - Filamentation is an efficient way to produce an intense and spectrally broad, but poorly stable, source for coherent control spectroscopy. We first described both theoretically and experimentally the filamentation and broadening of a 410 nm ultrashort laser pulse in Argon. By observing the theoretical and experimental spectral cross-correlation in the filament, we then show that the stability of the source can be improved. The Signal-to-Noise Ratio of the intensity inside the filament is increased up to 7 dB by its spectral filtering which provide a low noise broad spectrum source. PMID- 19550601 TI - Laser trapping of deformable objects. AB - We report the trapping and manipulation of bubbles in viscous glass melts through the use of a laser. This phenomenon is observed in bubbles tens of micrometers in diameter under illumination by low numerical aperture (NA = 0.55). Once the bubble was centered on the optical axis, it was trapped and followed a lateral relocation of the laser beam. This phenomenon is explained by modifications of the bubble's shape induced by axial heating and a decrease in surface tension. It is shown that formation of a concave dimple on the bubble's front surface explains the observed laser trapping and manipulation. This mechanism of laser trapping is expected to take place in other deformable materials and can also be used to remove bubbles from melts or liquids. For this technique to be effective, the alteration of the bubble's shape should be faster than its expulsion out of the laser's point of focus. PMID- 19550602 TI - Optical characterization of extremely small volumes of liquid in sub-micro-holes by simultaneous reflectivity, ellipsometry and spectrometry. AB - We have fabricated and characterized a lattice of submicron cone-shaped holes on a SiO(2)/Si wafer. Reflectivity profiles as a function of angle of incidence and polarization, phase shift and spectrometry are obtained for several fluids with different refractive indexes filling the holes. The optical setup allows measuring in the center of a single hole and collecting all data simultaneously, which can be applied for measuring extremely low volumes of fluid (in the order of 0.1 femtolitres) and label-free immunoassays, as it works as a refractive index sensor. A three layer film stack model is defined to perform theoretical calculations. PMID- 19550603 TI - Dynamic chirp control of all-optical format-converted pulsed data from a multi wavelength inverse-optical-comb injected semiconductor optical amplifier. AB - By spectrally and temporally reshaping the gain-window of a traveling-wave semiconductor optical amplifier (TWSOA) with a backward injected multi- or single wavelength inverse-optical-comb, we theoretically and experimentally investigate the dynamic frequency chirp of the all-optical 10GBit/s Return-to-Zero (RZ) data stream format-converted from the TWSOA under strong cross-gain depletion scheme. The multi-wavelength inverse-optical-comb injection effectively depletes the TWSOA gain spectrally and temporally, remaining a narrow gain-window and a reduced spectral linewidth and provide a converted RZ data with a smaller peak-to peak frequency chirp of 6.7 GHz. Even at high inverse-optical-comb injection power and highly biased current condition for improving the operational bit-rate, the chirp of the multi-wavelength-injection converted RZ pulse is still 2.1-GHz smaller than that obtained by using single-wavelength injection at a cost of slight pulse-width broadening by 1 ps. PMID- 19550604 TI - Discrimination between Doppler-shifted and non-shifted light in coherence domain path length resolved measurements of multiply scattered light. AB - We show a novel technique to distinguish between Doppler shifted and unshifted light in multiple scattering experiments on mixed static and dynamic media. With a phase modulated low coherence Mach- Zehnder interferometer, optical path lengths of shifted and unshifted light and path length dependent Doppler broadening are measured in a two-layer tissue phantom, with a superficial static layer of different thickness covering a semi-infinite dynamic medium having identical optical properties. No Doppler broadening is observed until a certain optical path length depending on the thickness of the superficial static layer. From the minimum optical path length corresponding to the Doppler-shifted light the thickness of the static layer that overlies the dynamic layer can be estimated. Validation of the experimentally determined thickness of the static layer is done with the Doppler Monte Carlo technique. This approach has potential applications in discriminating between statically and dynamically scattered light in the perfusion signal and in determining superficial burn depths. PMID- 19550605 TI - Nonlinear ellipse rotation modified Z-scan measurements of third-order nonlinear susceptibility tensor. AB - We present a method that combines the Z-scan technique with nonlinear ellipse rotation (NER) to measure third-order nonlinear susceptibility components. The experimental details are demonstrated, and a comprehensive theoretical analysis is given. The validity of this method is verified by the measurements of the nonlinear susceptibility tensor of a well-characterized liquid, CS2. PMID- 19550606 TI - Rigorous analysis of spheres in Gauss-Laguerre beams. AB - In this paper we develop a rigorous formulation of Gauss- Laguerre beams in terms of Mie scattering coefficients which permits us to quasi-analytically treat the interaction of a spherical particle located in the focal region of a possibly high numerical aperture lens illuminated by a Gauss-Laguerre beam. This formalism is used to study the scattered field as a function of the radius of a spherical scatterer, as well as the translation of a spherical scatterer through the Gauss Laguerre illumination in the focal plane. Knowledge of the Mie coefficients provides a deeper insight to understanding the scattering process and explaining the oscillatory behaviour of the scattered intensity distribution. PMID- 19550607 TI - Full range complex spectral domain optical coherence tomography without additional phase shifters. AB - We demonstrate a new full range complex spectral domain optical coherence tomography (FRC SD-OCT) method. Other than FRC SD-OCT systems reported in literature, which employed devices such as electro-/acousto optic modulators or piezo-driven mirrors providing the phase modulations necessary for retrieval of the complex-valued signal, the system presented works without any additional phase shifting device. The required phase shift is introduced by the galvanometer scanner used for transversally scanning the sample beam. By means of a slight displacement of the probe beam with respect to the scanning mirror's pivot axis, the sample arm length and thus the phase is continuously modulated as the beam is scanned in lateral direction. From such modulated spectral data, the complex valued data yielding a twofold increase of accessible depth range can be calculated using an algorithm based on the Hilbert transform. To demonstrate the performance of our method quantitative measurements of the suppression of mirror images as a function of induced phase shift were performed. In order to validate the FRC SD-OCT technique for high-speed imaging of biological tissue, we present full-range images of the human anterior chamber in vivo. PMID- 19550608 TI - Determination of Gamut Boundary Description for multi-primary color displays. AB - Displays with a larger color gamut to represent the images of the small color gamut are emphasized in the display development trend recently. Resulting from the vigorous development of Light Emitting Diodes (LEDs), the solutions to enlarge the color gamut which is formed a polygon area by adding multiple primary colors are possible, easier and inexpensive considerably. Therefore, how to determine the Gamut Boundary Description (GBD) plays a significant role for the applications of the multiple primary color displays, where the primaries form a convex polygon in CIE xy space. The paper proposed a method to construct the three-dimension color volume of GBD from the two-dimension polygon gamut area precisely regardless of that how many multiple primary colors the displays have. The method is examined in detail by the simulations and experiments, and proved it to fulfill from tri-primary color device to Nprimary color device. PMID- 19550609 TI - Excitation of a single hollow waveguide mode using inhomogeneous anisotropic subwavelength structures. AB - We propose and analyze a general approach for coupling a free space uniformly polarized beam to a desired hollow waveguide mode, thus enabling a single mode operation. The required spatial polarization state manipulation is achieved by use of inhomogeneous anisotropic subwavelength structures. Demonstration is obtained by coupling a linearly polarized CO(2) laser beam at a wavelength of 10.6 mum to the TE(01), TM(01), EH(11), EH(21), and EH(31) modes of a 300 mum diameter dielectric-coated hollow metallic waveguide. Full polarization and intensity analysis of the beam at the waveguide's inlet and outlet ports indicates a high coupling efficiency to a single waveguide mode. Finally, shaping the waveguide mode to a nearly diffraction limited linearly polarized beam and to a radially polarized vectorial vortex are also demonstrated. PMID- 19550611 TI - Analysis on fluorescence intensity reverse photonic phenomenon between red and green fluorescence of oxyfluoride nanophase vitroceramics. AB - An interesting fluorescence intensity reverse photonic phenomenon between red and green fluorescence is investigated. The dynamic range Sigma of intensity reverse between red and green fluorescence of Er(0.5)Yb(3):FOV oxyfluoride nanophase vitroceramics, when excited by 378.5nm and 522.5nm light respectively, is about 4.32x10(2). It is calculated that the phonon-assistant energy transfer rate of the electric multi-dipole interaction of {(4)G(11/2)(Er(3+))?(4)F(9/2)(Er(3+)), (2)F(7/2)(Yb(3+))?(2)F(5/2)(Yb(3+))} energy transfer of Er(0.5)Yb(3):FOV is around 1.380x10(8)s(-1), which is much larger than the relative multiphonon nonradiative relaxation rates 3.20x10(5)s(-1). That energy transfer rate for general material with same rare earth ion's concentration is about 1.194x10(5)s( 1). These are the reason to emerge the unusual intensity reverse phenomenon in Er(0.5)Yb(3):FOV. PMID- 19550610 TI - Luminescent images of single gold nanoparticles and their labeling on silica beads. AB - Luminescent Au nanoparticles were synthesized in a modified Brust method (average diameters of metal core = 1.6 nm). The fluorescence images were measured using scanning confocal microscopy and validated as compared with organic fluorophores. The metal particles were functionalized with succinimidyl ester terminated ligands and bound as fluorophores on surface-aminated silica beads to mimic labeling of biological functionalities. The labeled silica beads were shown to display bright signals and good photostability. Our results indicate that the luminescent metal nanoparticles can be employed as the probes to label the biological functionalities in developing molecule imaging agents. PMID- 19550612 TI - Back-scattered detection provides atomic-scale localization precision, stability, and registration in 3D. AB - State-of-the-art microscopy techniques (e.g., atomic force microscopy, scanning tunneling microscopy, and optical tweezers) are sensitive to atomic-scale (100 pm) displacements. Yet, sample drift limits the ultimate potential of many of these techniques. We demonstrate a general solution for sample control in 3D using back-scattered detection (BSD) in both air and water. BSD off a silicon disk fabricated on a cover slip enabled 19 pm lateral localization precision (Deltaf = 0.1-50 Hz) with low crosstalk between axes (99.9% and a mean group delay dispersion (GDD) of about -30 fs(2) with a theoretical GDD ripple of less than 0.5 fs(2) in the working spectral range. Deviations of the measured GDD from the calculated one are restricted to less than +/- 3 fs(2), limited by our measurement system. Simulations reveal that a dispersive delay line composed of 120 bounces off these mirrors introduces negligible distortion to a femtosecond pulse and largely preserves its contrast. The mirrors constitute an ideal tool for precision intracavity or extracavity dispersion control in the range of several thousand fs(2), particularly if pulses with high contrast are to be generated. PMID- 19550648 TI - Optical 'shorting wires'. AB - Connecting lumped circuit elements in a conventional circuit is usually accomplished by conducting wires that act as conduits for the conduction currents with negligible potential drops. More challenging, however, is to extend these concepts to optical nanocircuit elements. Here, following our recent development of optical lumped circuit elements, we show how a special class of nanowaveguides formed by a thin core with relatively large (positive or negative) permittivity surrounded by a thin concentric shell with low permittivity may provide the required analogy to 'wires' for optical nano-circuits. PMID- 19550649 TI - Coexistence of total internal reflexion and bandgap modes in solid core photonic bandgap fibre with intersticial air holes. AB - In this article, we deal with new properties of a Solid Core Photonic Bandgap (SC PBGF) fiber with intersticial air holes (IAHs) in its transverse structure. It has been shown recently, that IAH enlarges its bandgaps (BG), compared to what is observed in a regular SC-PBGF. We shall describe the mechanisms that account for this BG opening, which has not been explained in detail yet. It is then interesting to discuss the role of air holes in the modification of the Bloch modes, at the boundaries of the BG. In particular, we will use a simple method to compute the exact BG diagrams in a faster way, than what is done usually, drawing some parallels between structured fibers and physics of photonic crystals. The very peculiar influence of IAHs on the upper/lower boundaries of the bandgaps will be explained thanks to the difference between mode profiles excited on both boundaries, and linked to the symmetry / asymmetry of the modes. We will observe a modification of the highest index band (n(FSM)) due to IAHs, that will enable us to propose a fiber design to guide by Total Internal Reflection (TIR) effect, as well as by a more common BG confinement. The transmission zone is deeply enlarged, compared to regular photonic bandgap fibers, and consists in the juxtaposition of (almost non overlapping) BG guiding zones and TIR zone. PMID- 19550650 TI - Light scattering by a nanoparticle and a dipole placed near a dielectric surface covered by a thin metallic film. AB - On the basis of Maxwell's equations a light scattering system of axial symmetry is considered, which consists of a nanoparticle, a dipole and a metal film (covering a dielectric support). Nanoparticle (NP) and dipole are situated on an axis of symmetry and the dipole is oriented along the axis and placed between film and nanoparticle. The field enhancement factor F and dipole energy flux D are calculated by the Green's function method: the initial system of Maxwell's equations is reduced to a system of boundary integral equations, and solutions are obtained by the boundary element method. Illumination of the scattering system by a radially polarized Bessel light beam causes a field enhancement in the vicinity of the film surface. The metallic NP closely placed at the film surface acts as nano-antenna. Surface plasmons excited in the particle and film convert the incident propagating EM field into non-propagating evanescent near field. Then the field is confined and strongly enhanced in a particle/film gap. The enhancement of Raman radiation depends on many factors: size and shape of NP, permittivities of all materials, light wavelength, film thickness, angle of light beam, and - very strongly - on the gap distance. The field enhancement in a gap ~1 nm can be 10(3) and more and the Raman radiation enhancement factor can reach huge values ~10(10)-10(12). Whereas for small nanoparticles the field enhancement factor F and the dipole energy flux D do not depend on the direction of the exciting beam and on the angle of emission, a strong influence is found for extended particles. This influence is plausibly explained by a larger overlap between the electric field of the exciting beam or the emitted radiation field with the near field distribution of the nanoparticle leading to higher F and D values, respectively. PMID- 19550651 TI - Roughness-induced radiation losses in optical micro or nanofibers. AB - Roughness-induced radiation losses in optical micro or nanofibers (MNFs) are investigated using an induced-current model. Loss coefficients of silica, phosphate, tellurite and silicon MNFs with sinusoidal deformations on their surfaces are numerically calculated with respect to typical parameters of the guiding system. Interesting phenomena such as the existence of the loss minima at specific perturbation periods are observed. Results presented in this work may be generalized to all kinds of surface deformation and may provide useful guidelines for both estimating and tailoring waveguiding properties of MNFs. PMID- 19550652 TI - Statistical speckle study to characterize scattering media: use of two complementary approaches. AB - Speckle produced by strongly-scattering media contains information about its optical properties. Statistical speckle study allows discrimination between media and enables one to characterize any change. Two approaches of the speckle phenomenon are used in the measurement of speckle produced by monodisperse polystyrene microspheres in solution and mixtures of them: a stochastic approach based on the fractional Brownian motion and a classical frequential approach based on speckle size measurement. In this paper, we introduce an approach that contains the multi-scale aspect of the speckle; therefore it provides more information on the medium than the speckle dimension. The obtained results show that the stochastic approach allows a better samples discrimination than the classical frequential approach. PMID- 19550653 TI - Giant enhancement of band edge emission based on ZnO/TiO(2) nanocomposites. AB - Enhancement of band edge emission of ZnO nanorods up to a factor of 120 times has been observed in the composite consisting of ZnO nanorods and TiO(2) nanoparticles, while the defect emission of ZnO nanorods is quenched to noise level. Through a detailed investigation, it is found that the large enhancement mainly arises from fluorescence resonance energy transfer between the band edge transition of ZnO nanorods and TiO(2) nanoparticles. Our finding opens up new possibilities for the creation of highly efficient solid state emitters. PMID- 19550654 TI - Enhanced optical absorptance of metals using interferometric femtosecond ablation. AB - The enhanced optical absorptance in metals was recently demonstrated using femtosecond laser-induced surface structuring. This structuring was obtained by simply focusing the light to the sample surface. Here we demonstrate more efficient absorptance enhancement using interferometric ablation. This interferometric ablation technique produces deeper surface structures and, consequently, higher absorption than structures obtained by just focusing the light to the surface. We also show the measured reflectance spectra over visible region for unaltered and structured stainless steel and copper samples. PMID- 19550655 TI - Stabilized dual-wavelength erbium-doped dual-ring fiber laser. AB - A stable dual-wavelength erbium-doped fiber (EDF) compound ring laser using a double-ring filter (DRF) is proposed and demonstrated experimentally. By using a ring filter incorporated within the ring cavity, the fiber laser can lase two wavelengths simultaneously. The dual-wavelength output exhibits a good performance having the optical side-mode suppression ratios (SMSRs) of 45.3 and 46.3 dB and output powers of -8.1 and -7.1 dBm, respectively. In addition, the optical output stabilities of the ring laser have been also discussed. PMID- 19550656 TI - Laser damage resistance of RbTiOPO(4): evidence of polarization dependent anisotropy. AB - Nanosecond-laser induced damage of RbTiOPO(4) crystals (RTP) has been studied at 1064 nm as a function of propagation direction and polarization orientation. A significant difference in the Laser Induced Damage Threshold (LIDT) was observed for x-cut and y-cut crystals in Pockels cell configuration, where the light propagation direction is along the x and y axes of the crystal respectively. In Pockels cell configuration the polarization is oriented at 45? with respect to the z-axis of the crystal. Experiments with the polarization oriented parallel to the principal axes of the crystal pointed out the importance of the polarization direction for the LIDT whereas the propagation direction did not significantly influence the LIDT. Comparison of the experimental data with a simple model reveals the influence of frequency doubling on the LIDT in Pockels cell configuration. In the case of the y-cut Pockels cell, the generation of frequency doubled light causes an LIDT below the LIDT of x and z-polarized light at the fundamental wavelength. PMID- 19550657 TI - Devil's lenses. AB - In this paper we present a new kind of kinoform lenses in which the phase distribution is characterized by the "devil's staircase" function. The focusing properties of these fractal DOEs coined devil's lenses (DLs) are analytically studied and compared with conventional Fresnel kinoform lenses. It is shown that under monochromatic illumination a DL give rise a single fractal focus that axially replicates the self-similarity of the lens. Under broadband illumination the superposition of the different monochromatic foci produces an increase in the depth of focus and also a strong reduction in the chromaticity variation along the optical axis. PMID- 19550658 TI - Coupling of optical lumped nanocircuit elements and effects of substrates. AB - We present here an analytical quasi-static circuit model for the coupling among small nanoparticles excited by an optical electric field in the framework of the optical lumped nanocircuit theory [N. Engheta, A. Salandrino, and A. Alu, Phys. Rev. Lett. 95, 095504 (2005)]. We derive how coupling effects may affect the corresponding nanocircuit model by adding lumped controlled sources that depend on the optical voltages applied on the coupled particles as coupled lumped elements. With the same technique, we may model the presence of a substrate located underneath the nanocircuit elements, relating its presence to the coupling with a properly modeled image nanoparticle. These results are of importance in the understanding and the design of complex optical nanocircuits at infrared and optical frequencies. PMID- 19550659 TI - Electromagnetic equivalent model for phase conjugate mirror based on the utilization of left-handed material. AB - An electromagnetic equivalent model for the phase conjugate mirror (PCM) is proposed in this paper. The model is based on the unique property of the isotropic left-handed material (LHM) - the ability of LHM to reverse the phase factors of propagative waves. We show that a PCM interface can be substituted with a LHM-RHM (right-handed material) interface and associated image sources and objects in the LHM. This equivalent model is fully equivalent in the treatment of propagative wave components. However, we note that the presence of evanescent wave components can lead to undesirably surface resonance at the LHM-RHM interface. This artefact can be kept well bounded by introducing a small refractive index mismatch between the LHM and RHM. We demonstrate the usefulness of this model by modelling several representative scenarios of light patterns interacting with a PCM. The simulations were performed by applying the equivalent model to a commercial finite element method (FEM) software. This equivalent model also points to the intriguing possibility of realizing some unique LHM based systems in the optical domain by substituting a PCM in place of a LHM-RHM interface. PMID- 19550660 TI - Optical characterization of a GaAs/In(0.5)(AlxGa(1-x))(0.5)P/GaAs heterostructure cavity by piezoreflectance spectroscopy. AB - Optical properties of a lattice matched GaAs/In(0.5)(AlxGa(1-x))(0.5)P/GaAs heterostructure cavity have been characterized using piezoreflectance (PzR) measurements in the temperature range between 20 and 300 K. The measurements were carried out in the energy range of 1.3-6 eV. The PzR spectra of In(0.5)(AlxGa(1 x))(0.5)P at 20 and 300K clearly show a lot of interband transition features present at energies above the band edge. There is also a feature of interference fringes oscillations observed in each PzR spectrum below band gap E0 of In(0.5)(AlxGa(1-x))(0.5)P. The oscillation period in between the PzR interference fringes can be utilized to determine the index of refraction (n) for the In(0.5)(AlxGa(1-x))(0.5)P at different temperatures. The Al composition x of In(0.5)(AlxGa(1-x))(0.5)P can be estimated from the evaluation value of E(0) by PzR. The obtained Al composition of x=0.691 is in good agreements with the original design for growing the quaternary compound. Electronic band structure of In(0.5)(AlxGa(1-x))(0.5)P is determined by the interband transitions from PzR. The temperature variations of the transition energies and index of refraction n for the In(0.5)(AlxGa(1-x))(0.5)P are analyzed discussed. The PzR is proven to be very sensitive in determining the optical parameters in III-V GaAs/InAlGaP/GaAs Fabry-Perot system. PMID- 19550661 TI - Second-harmonic emission from sub-wavelength apertures: Effects of aperture symmetry and lattice arrangement. AB - We measure second-harmonic generation from arrays of sub-wavelength apertures in transmission using fundamental input at 800 nm. Lattice arrangements include disordered, Penrose (quasi-periodic or aperiodic), and square (periodic). Strong angular dependence of SHG is observed, with maxima located at angular positions that roughly correspond to incidence angles of extraordinary optical transmission (EOT) for the fundamental. In addition, even at incidence normal to the sample, strong secondary maxima are observed at off-normal scattering angles for the arrangements with higher degree of order. Breaking the inversion symmetry of the aperture allows second harmonic peaks at normal incidence and detection. These measurements help to resolve the role that symmetry plays in second-harmonic generation from arrays of apertures. PMID- 19550662 TI - Mirror cavity MMI coupled photonic wire resonator in SOI. AB - We propose a new waveguide resonator device with a mirror cavity and a multimode interference (MMI) coupler. We present simulation results for the silicon wire MMI coupler with suppressed reflections and its use as a coupling element in the resonator cavity, built on the silicon-on-insulator waveguide platform. Tapering structures used in the reflection suppression were optimized, and the wavelength dependency of a conventional MMI was compared to that of the MMI with reflection suppression. Equations relating the power transfer of the two-mirror MMI-coupled resonator and quality factor were derived. The device was also studied using finite difference time domain simulation by both pulse and continuous wave excitation. The resonator does not require bend waveguides, it has the advantages of having no bend loss and a compact layout. The resonator device has a very small footprint of 3 mum x 30 mum, and a quality factor of 516. PMID- 19550663 TI - Real time sensor for monitoring oxygen in radio-frequency plasma applications. AB - Real time closed loop control of plasma assisted semiconductor manufacturing processes has received significant attention in recent years. Therefore we have developed and tested a customized optical sensor based on buffer gas (argon) actinometry which has been used to determine relative densities of atomic and molecular oxygen in an Ar/O(2) radio-frequency ICP chamber. The operation and accuracy of our optical sensor compared favorably with a high resolution commercial spectrometer but at lower cost and exhibited improved actinometric performance over a low resolution commercial spectrometer. Furthermore, threshold tests have been performed on the validity of buffer gas based actinometry in Ar/O(2) ICP plasmas where Ar is no longer a trace gas through Xe actinometry. The plasma conditions for which this customized optical sensor can be used for closed loop control have been established. PMID- 19550664 TI - Photochromism and two-photon luminescence of Ag-TiO(2) granular composite films activated by near infrared ps/fs pulses. AB - We reported photochromism and largely enhanced visible two-photon luminescence (TPL) of Ag-TiO(2) granular composite films by using ps/fs laser at the wavelength of 800 nm. Three types of photochromism spectra were observed when the Ag atom fraction are less than, comparable to and larger than the percolation threshold. The strong surface-plasmon-resonance enhanced visible TPL emissions near Ag(2)O transition band from the photoactivated Ag-TiO(2) samples were also observed. Furthermore, we found that the TPL intensity saturatedly increased while the absorbance at 800 nm exponentially decreased with the same rate as the increasing of photoactivation time, which means that both photochromism and TPL of Ag-TiO(2) composite films are originated from the photo-oxidation of Ag to Ag(+). These observations exhibit the multifunctional features of Ag nanoparticle materials. PMID- 19550665 TI - Quasi-two-level Yb:KYW laser with a volume Bragg grating. AB - Using a volume Bragg grating as input coupler, we demonstrate an Yb:KYW laser with a very small quantum defect (1.6%) and an output power of 3.6 W. The laser was longitudinally diode-pumped at 982 nm and the laser wavelength was determined by the grating to 998 nm, with a laser bandwidth of 10 GHz (33 pm). Due to the low quantum defect, the laser should be readily scalable to 20 W or more without critical thermal effects. PMID- 19550666 TI - Origin of coupling to antisymmetric modes in arc-induced long-period fiber gratings. AB - We study the origin of antisymmetric perturbation of the fiber in arc-induced long-period gratings that couple the core mode into the antisymmetric cladding modes. We demonstrate that this perturbation is caused by the temperature gradient in the fiber, which is induced, in turn, by the temperature gradient in the arc discharge. The reproducibility of the process of the grating inscription is higher when the fiber is placed in a region with larger temperature gradient. PMID- 19550667 TI - Generation of hard X-rays using an ultrafast fiber laser system. AB - We report the first hard X-ray source driven by a femtosecond fiber laser. The high energy fiber CPA system incorporated a 65mum LMA fiber amplifying stage which provided 300-fs recompressed pulses and diffraction limited beam quality with M(2) < 1.07. A deformable mirror was used to optimize the wavefront and the spot size was focused down to 2.3 mum with an f/1.2 paraboloidal mirror. 50muJ was deposited on the nickel target with 2x10(15)-W/cm(2) focal intensity and a distinctive Ni K(alpha)-line (7.48 keV) emission was measured with 5x10(-8) energy conversion efficiency. PMID- 19550668 TI - Surface profilometry with composite interferometer. AB - We proposed and demonstrated a low-cost optical system for surface profilometry with nanometer-resolution. The system is based on a composite interferometer consisting of a Michelson interferometer and a Mach-Zehnder interferometer. With the proposed phase compensating mechanism, the phase deviation due to the instability of the optical delay system and environmental perturbation can be compensated simultaneously. The system can perform a wide-field imaging in the millimeter range and a measurement with the axial resolution within +/-5 nm without special shielding and protection of the system as well as any special preparation of the sample. PMID- 19550669 TI - Long-distance distribution of time-bin entangled photon pairs over 100 km using frequency up-conversion detectors. AB - We report an experimental demonstration of the distribution of time-bin entangled photon pairs over 100 km of optical fiber. In our experiment, 1.5-mum non degenerated time-bin entangled photon pairs were generated with a periodically poled lithium niobate (PPLN) waveguide by using the parametric down conversion process. Combining this approach with ultra-low-loss filters to eliminate the pump light and separate signal and idler photons, we obtained an efficient entangled photon pair source. To detect the photons, we used single-photon detectors based on frequency up-conversion. These detectors operated in a non gated mode so that we could use a pulse stream of time correlated entangled photon pairs at a high repetition frequency (1 GHz). Using these elements, we distributed time-bin entangled photon pairs over 100 km of dispersion shifted fiber and performed a two-photon interference experiment. We obtained a coincidence fringe of 81.6% visibility without subtracting any background noise, such as accidental coincidence or dark count, which was good enough to violate Bell's inequality. Thus, we successfully distributed time-bin entangled photon pairs over 100 km. PMID- 19550670 TI - 31 GHz Ge n-i-p waveguide photodetectors on Silicon-on-Insulator substrate. AB - We report on evanescently coupled Ge waveguide photodetectors that are grown on top of Si rib waveguides. A Ge waveguide detector with a width of 7.4mum and length of 50 mum demonstrated an optical bandwidth of 31.3 GHz at -2V for 1550nm. In addition, a responsivity of 0.89 A/W at 1550 nm and dark current of 169 nA were measured from this detector at -2V. A higher responsivity of 1.16 A/W was also measured from a longer Ge waveguide detector (4.4 x 100 mum2), with a corresponding bandwidth of 29.4 GHz at -2V. An open eye diagram at 40 Gb/s is also shown. PMID- 19550671 TI - Transverse particle dynamics in a Bessel beam. AB - Spatially periodic optical fields can be used to sort dielectric microscopic particles as a function of size, shape or refractive index. In this paper we elucidate through both theory and experiment the behavior of silica microspheres moving under the influence of the periodic optical field provided by a Bessel beam. We compare two different computational models, one based on Mie scattering, the other on geometrical ray optics and find good qualitative agreement, with both models predicting the existence of distinct size-dependent phases of particle behavior. We verify these predictions by providing experimental observations of the individual behavioral phases. PMID- 19550672 TI - Monte Carlo simulation of propagation of a short light beam through turbulent oceanic flow. AB - We use Monte Carlo time-dependent simulations of light pulse propagation through turbulent water laden with particles to investigate the application of Multiple Field Of View (MFOV) lidar to detect and characterize oceanic turbulence. Inhomogeneities in the refractive index induced by temperature fluctuations in turbulent ocean flows scatter light in near-forward angles, thus affecting the near-forward part of oceanic water scattering phase function. Our results show that the oceanic turbulent signal can be detected by analyzing the returns from a MFOV lidar, after re-scaling the particulate back scattering phase function. PMID- 19550673 TI - EUV reflectance and scattering of Mo/Si multilayers on differently polished substrates. AB - Highly reflective Molybdenum/Silicon multilayer mirrors for 13.5 nm are characterized at-wavelength using a new laboratory size measurement system for EUV reflectance and scattering. Roughness analysis before and after coating by Atomic Force Microscopy indicates roughness enhancement as well as smoothing effects during thin film growth. The impact of the substrate finish and the deposition process onto the scattering distribution and scatter losses with regard to the specular reflectance is analyzed. PMID- 19550674 TI - Single-shot compressive spectral imaging with a dual-disperser architecture. AB - This paper describes a single-shot spectral imaging approach based on the concept of compressive sensing. The primary features of the system design are two dispersive elements, arranged in opposition and surrounding a binary-valued aperture code. In contrast to thin-film approaches to spectral filtering, this structure results in easily-controllable, spatially-varying, spectral filter functions with narrow features. Measurement of the input scene through these filters is equivalent to projective measurement in the spectral domain, and hence can be treated with the compressive sensing frameworks recently developed by a number of groups. We present a reconstruction framework and demonstrate its application to experimental data. PMID- 19550675 TI - Coherent anti-Stokes Raman scattering microscopy using photonic crystal fiber with two closely lying zero dispersion wavelengths. AB - We demonstrate coherent anti-Stokes Raman scattering (CARS) microscopy of lipid rich structures using a single unamplified femtosecond Ti:sapphire laser and a photonic crystal fiber (PCF) with two closely lying zero dispersion wavelengths (ZDW) for the Stokes source. The primary enabling factor for the fast data acquisition (84 micros per pixel) in the proof-of-principle CARS images, is the low noise supercontinuum (SC) generated in this type of PCF, in contrast to SC generated in a PCF with one ZDW. The dependence of the Stokes pulse on average input power, pump wavelength, pulse duration and polarization is experimentally characterized. We show that it is possible to control the spectral shape of the SC by tuning the pump wavelength of the input pulse and the consequence for CARS microscopy is discussed. PMID- 19550676 TI - Chromatic dispersion and polarization mode dispersion monitoring for multi-level intensity and phase modulation systems. AB - We demonstrate chromatic dispersion (CD) and polarization mode dispersion (PMD) monitoring techniques via simulation and experiment for 2- and 4-level intensity modulated as well as phase-modulated optical systems. Degree of polarization (DOP) measurement for monitoring PMD up to 100-ps and clock tone measurement for monitoring CD up to 720-ps/nm are demonstrated in 10-Gsymbol/s non-return-to-zero (NRZ) and return-to-zero (RZ) systems. Analysis on dynamic range and monitoring window shows that careful consideration and characterization are necessary when applying these monitoring techniques to multi-level systems. PMID- 19550677 TI - Characterization of near-infrared low energy ultra-short laser pulses for portable applications of laser induced breakdown spectroscopy. AB - We report on the delivery of low energy ultra-short (<1 ps) laser pulses for laser induced breakdown spectroscopy (LIBS). Ultra-short pulses have the advantage of high peak irradiance even at very low pulse energies. This opens the possibility to use compact, rare-earth doped fiber lasers in a portable platform for point detection applications using LIBS for elemental analysis. The use of low energy ultra-short pulses minimizes the generation of a broad continuum background in the emission spectrum, which permits the use of non-gated detection schemes using very simple and compact spectrometers. The pulse energies used to produce high-quality LIBS spectra in this investigation are some of the lowest reported and we investigate the threshold pulse requirements for a number of near IR pulse wavelengths (785-1500 nm) and observe that the pulse wavelength has no effects on the threshold for observation of plasma emission or the quality of the emission spectra obtained. PMID- 19550678 TI - Volumetric motility-contrast imaging of tissue response to cytoskeletal anti cancer drugs. AB - Microscopic imaging of cellular motility has recently advanced from two dimensions to three dimensions for applications in drug development. However, significant degradation in resolution occurs with increasing imaging depth, limiting access to motility information from deep inside the sample. Here, digital holographic optical coherence imaging is adapted to allow visualization of motility in tissue at depths inaccessible to conventional motility assay approaches. This method tracks the effect of cytoskeletal anti-cancer drugs on tissue inside its natural three-dimensional environment using time-course measurement of motility within tumor tissue. PMID- 19550679 TI - Petal-like modes in Porro prism resonators. AB - A new approach to modeling the spatial intensity profile from Porro prism resonators is proposed based on rotating loss screens to mimic the apex losses of the prisms. A numerical model based on this approach is presented which correctly predicts the output transverse field distribution found experimentally from such resonators. PMID- 19550680 TI - Electrically switchable and optically rewritable reflective Fresnel zone plate in dye-doped cholesteric liquid crystals. AB - This work demonstrates a reflective Fresnel zone plate based on dye-doped cholesteric liquid crystals (DDCLC) using the photo-induced realignment technique. Illumination of a DDCLC film with a laser beam through a Fresnel-zone plate mask yields a reflective lens with binary-amplitude structures - planar and focal conic textures, which reflect and scatter probed light, respectively. The formed lens persists without any external disturbance, and its focusing efficiency, analyzed using circularly polarized light, is ~ 23.7%, which almost equals the measured diffraction efficiency of the used Fresnel-zone-plate mask (~ 25.6%). The lens is thermally erasable, rewritable and switchable between focusing and defocusing states, upon application of a voltage. PMID- 19550681 TI - Shape based Monte Carlo code for light transport in complex heterogeneous Tissues. AB - A Monte Carlo code for the calculation of light transport in heterogeneous scattering media is presented together with its validation. Triangle meshes are used to define the interfaces between different materials, in contrast with techniques based on individual volume elements. This approach allows to address realistic problems in a flexible way. A hierarchical spatial organisation enables a fast photon-surface intersection test. The application of the new environment to evaluate the impact of the trabecular structure of bone on its optical properties is demonstrated. A model of the trabecular micro structure recovered from microCT data was used to compute light distribution within tissue. Time resolved curves across a spherical bone volume were computed. The work presented enables simulation of radiative transport in complex reality-based models of tissue and serves as a powerful, generic tool to study the effect of heterogeneity in the field of biomedical optics. PMID- 19550682 TI - Electroluminescence of nanopatterned silicon with carbon implantation and solid phase epitaxial regrowth. AB - Electroluminescence at 1.28mum is observed in a nanopatterned silicon test structure that has been subjected to carbon implantation followed by solid-phase epitaxial regrowth for recrystalization. The sub-bandgap luminescence comes from a di-carbon complex known as 'G center'. Enrichment of silicon with carbon atoms has been achieved in a procedure consisting of two implantations and solid-phase epitaxial regrowth. Nanopatterning was done using an anodized aluminum oxide membrane as a mask for reactive ion etching. Along with the electroluminescence, an enhanced photoluminescence was measured. PMID- 19550683 TI - Improved analysis on the signal property of computational integral imaging system. AB - In this paper, we introduce an improved signal analysis of the computational integral imaging (CII) system having a pickup process of three-dimensional object and a volumetric computational reconstruction (VCR) process. We propose a signal model for the CII system. From the signal model and its analysis, we can define a granular noise caused by the non-uniform overlapping. We also analyze the characteristics of the granular noise. According to our model and analysis, there is a condition that the granular noise cancels out. To show the feasibility of our model, the preliminary experiments are carried out and the result is presented. This is the first time, to our knowledge, that a signal model for the analysis of CII systems is provided. PMID- 19550684 TI - Statistical analysis of measured free-space laser signal intensity over a 2.33 km optical path. AB - Experimental research is conducted to determine the characteristic behavior of high frequency laser signal intensity data collected over a 2.33 km optical path. Results focus mainly on calculated power spectra and frequency distributions. In addition, a model is developed to calculate optical turbulence intensity (C(n)/2) as a function of receiving and transmitting aperture diameter, log-amplitude variance, and path length. Initial comparisons of calculated to measured C(n)/2 data are favorable. It is anticipated that this kind of signal data analysis will benefit laser communication systems development and testing at the U.S. Army Research Laboratory (ARL) and elsewhere. PMID- 19550685 TI - Micro Fabry-Perot interferometers in silica fibers machined by femtosecond laser. AB - Micro Fabry-Perot (F-P) interferometers (MFPIs) are machined in a single-mode fiber (SMF) and a photonic crystal fiber (PCF) by using a near-infrared femtosecond laser, respectively. The strain and temperature characteristics of the two MFPIs with an identical cavity length are investigated and the experimental results show that the strain sensitivity of the PCF-based MFPI is smaller than that of the SMF-based MFPI due to their different waveguide structures, while the two MFPIs have close temperature sensitivities due to their similar host materials. These MFPIs in silica fibers are compact, stable, inexpensive, capable for mass-production and easy fabrication, offering great potentials for wide sensing applications. PMID- 19550686 TI - Plasmonic nanoclusters: a path towards negative-index metafluids. AB - We introduce the concept of metafluids-liquid metamaterials based on clusters of metallic nanoparticles which we will term Artificial Plasmonic Molecules (APMs). APMs comprising four nanoparticles in a tetrahedral arrangement have isotropic electric and magnetic responses and are analyzed using the plasmon hybridization (PH) method, an electrostatic eigenvalue equation, and vectorial finite element frequency domain (FEFD) electromagnetic simulations. With the aid of group theory, we identify the resonances that provide the strongest electric and magnetic response and study them as a function of separation between spherical nanoparticles. It is demonstrated that a colloidal solution of plasmonic tetrahedral nanoclusters can act as an optical medium with very large, small, or even negative effective permittivity, epsilon(eff), and substantial effective magnetic susceptibility, Chi(eff) = mu(eff) -1, in the visible or near infrared bands. We suggest paths for increasing the magnetic response, decreasing the damping, and developing a metafluid with simultaneously negative epsilon(eff) and mu(eff). PMID- 19550687 TI - Design of spherically corrected, achromatic variable-focus liquid lenses. AB - A design method for correcting chromatic as well as spherical aberrations of variable-focus, multi-chamber liquid lenses is described. By combining suitable optical liquids with appropriate radii of the liquid's interfaces, liquid lenses with superior, diffraction-limited resolution over a wide focal tuning range are possible. For an infinite object distance, the analytic thin-lens approximation of an achromatic positive/negative varifocal liquid lens is derived and the obtained results are compared with ray-traced optimized designs which consider finite thicknesses and rigid cover glasses. As a design example, the optical performance of a 4mm-diameter positive/negative f /3.6 achromatic liquid lens is given in detail. PMID- 19550688 TI - Coherence collapse and low-frequency fluctuations in quantum-dash based lasers emitting at 1.57 mum. AB - Optical feedback tolerance is experimentally investigated on a 600-mum-long quantum-dash based Fabry-Perot laser emitting at 1.57mum. While quantum-dashes are structurally intermediate to quantum-wells and quantum-dots, the observed behaviour is distinctly like that of a quantum-well based laser but with greater stability. Coherence collapse and low-frequency fluctuation regimes are observed and are reported here. The onset of the coherence collapse regime is experimentally determined and is found to vary from -29 dB to -21 dB external feedback level when increasing the current from twice to nine times the threshold current. PMID- 19550689 TI - Counterpropagating nondiffracting beams through reflection gratings. AB - We investigate the counterpropagation of paraxial non-diffracting optical beams through a medium hosting a bulk reflection grating in the quasi-Bragg matching condition. The impact of the relative magnitude of the Bragg detuning parameter and the grating depth on the plane wave dispersion relation allows us to identify three distinct regimes where counterpropagation and interaction of nondiffracting beams show qualitatively different features, encompassing longitudinally invariant, periodic or exponential intensity profiles. In one of the identified regimes the dispersion relation is not monotonic and the consequent "longitudinal degeneracy" allows the investigation of new class of nondiffracting beams characterized by a double spectral ring profile. PMID- 19550690 TI - Linear and nonlinear optical properties of Hafnium-doped lithium-niobate crystals. AB - Measurements of birefringence, second-harmonic phase-matching conditions, and nonlinear coefficient d(31) are performed for a set of Hafnium-doped congruent lithium niobate (Hf:cLN) crystals as functions of dopant concentration. The data highlight that the threshold concentration, above which there is a change in the Hf incorporation mechanism, is slightly above 2mol% and that, up to this value of concentration, the efficiency of nonlinear processes is not affected by the dopant insertion. Combining these results with those already present in literature, Hf:cLN crystals appear to be very promising candidates for the development of photorefractivity-free wavelength converters working at room temperature. PMID- 19550691 TI - Localized surface plasmon nanolithography with ultrahigh resolution. AB - A localized surface plasmon nanolithography (LSPN) technique is proposed and demonstrated to produce patterns with a sub-20nm line width. High transmission efficiency is realized by adjusting the period of grating. The well-regulated grating structures in metallic mask are employed to excite surface plasmon polaritons (SPPs) on the illuminated side. The SPP waves propagate toward the tip along the taper surfaces which cause most of energy accumulation at the tip and give rise to high local field enhancements in a near-field region around the tip. The amplitude of local electric field intensity is quite large and the line width can be confined within sub-20nm, at the same time, the contrast and spatial resolution are greatly enhanced, which can facilitate nanolithography efficiently with simple ultraviolet light sources. PMID- 19550692 TI - Label-free optical imaging of mitochondria in live cells. AB - The far-field optical imaging of mitochondria of live cells without the use of any label is demonstrated. It uses a highly sensitive photothermal method and has a resolution comparable to confocal fluorescence setups. The morphological states of mitochondria were followed under different physiological treatments, and the role of cytochrome c was ruled out as the main origin of the photothermal signals. This label free optical method provides a high contrast imaging of live mitochondria and should find many applications in biosciences. PMID- 19550693 TI - Effectiveness of recycling light in ultra-bright short-arc discharge lamps. AB - Recycling light back into a plasma lamp's radiant zone can enhance its radiance. Measurements are reported for the effectiveness, spectral properties and modified plasma radiance maps that result from light recycling with a specular hemispherical mirror in commercial 150 W ultrabright Xenon short-arc discharge lamps, motivated by projection, biomedical and high-temperature furnace applications. For certain spectral windows and plasma arc regions, radiance can be heightened by up to 70%. However, the overall light recycling efficiency is reduced to about half this value due to lamp geometry. The manner in which light plasma interactions affect light recycling efficacy is also elucidated. PMID- 19550694 TI - Tomographic reconstruction of weak, replicated index structures embedded in a volume. AB - Measurements of weak, embedded index structures are important for material characterization of photopolymers, glass and other optical materials as well as for characterization of fabricated structures such as waveguides. We demonstrate an optical diffraction tomography system capable of measuring deeply-buried, weak, fabricated index structures written in a homogeneous volume. High-fidelity cross sections of these weak index structures are constructed by replicating the structure to be measured to form a diffraction grating. The coherent addition of scattering from each of these objects increases the sensitivity of the imaging system. Measurements are made in the far field, without the use of lenses, eliminating phase aberration errors through thick volumes. PMID- 19550695 TI - Analysis of frequency offset in the frequency stabilization of semiconductor laser based on frequency dithering technique. AB - We investigate the frequency offset in the frequency stabilization of a semiconductor laser based on a frequency-dithering technique. An analytical model is presented to describe the effects of the amplitude modulation and the phase delay between the amplitude and frequency modulation on the frequency stabilization. We also experimentally and analytically show that the frequency offset could be reduced by using an appropriate phase-sensitive detection. PMID- 19550696 TI - Negative beam displacements from negative-index photonic metamaterials. AB - It is well known that refraction of light at interfaces leads to a beam displacement for oblique incidence of light onto a slab of material. In ray optics and for homogeneous isotropic materials, the sign of this beam displacement is strictly identical to the sign of the refractive index. Our numerical calculations reveal negative beam displacements from state-of-the-art double-fishnet-type photonic metamaterials. This holds true for the "main" polarization corresponding to a negative phase velocity for normal incidence as well as for the "secondary" polarization with positive phase velocity. To understand and interpret these results, we also present exact analytical calculations for thin metal films showing that, in wave optics, the sign of the beam displacement (i.e., the sign of refraction) is generally not identical to the sign of the refractive index. PMID- 19550697 TI - Reduction of polarization-induced performance degradation in WDM PON utilizing MQW-SLD-based broadband source. AB - We report on the reduction of polarization-induced performance degradation in WDM PON utilizing MQW-SLD-based ASE source for injection locking to FPLD. The results show that, to suppress the polarization-induced Q penalty sufficiently less than 0.5 dB, the MQW-SLD output should be depolarized within the locking range of the wavelength-locked FPLD. PMID- 19550698 TI - Optimization of all-optical EDFA-based Sagnac-interferometer switch. AB - We perform optimization of all-optical EDFA-based Sagnac - interferometer switch through an analytical model and numerical simulations by solving nonlinear Schrodinger equations. The effects of the performance of EDFA on the bit rate and the switching power are investigated for all-optical switch based on self-phase or cross-phase modulation. The simulated results show that ultra-low switching power (<1mW) all-optical switch for 40 Gb/s data can be realized by properly selecting the length of highly nonlinear photonic crystal fiber and adjusting the performance of EDFA. PMID- 19550699 TI - Quantum interference fringes beating the diffraction limit. AB - Spatially formed two-photon interference fringes with fringe periods smaller than the diffraction limit are demonstrated. In the experiment, a fringe formed by two photon NOON states with wavelength lambda=702.2 nm is observed using a specially developed near-field scanning optical microscope probe and two-photon detection setup. The observed fringe period of 328.2 nm is well below the diffraction limit (351 nm = lambda /2). Another experiment with a path-length difference larger than the coherent length of photons confirms that the observed fringe is due to two-photon interference. PMID- 19550700 TI - Simple coherent polarization manipulation scheme for generating high power radially polarized beam. AB - We present a simple novel scheme that converts a Gaussian beam into an approximated radially polarized beam using coherent polarization manipulation together with Poynting walk-off in birefringent crystals. Our scheme alleviates the interferometric stability required by previous schemes that implemented this coherent mode summation using Mach-Zehnder-like interferometers. A symmetrical arrangement of two walk-off crystals with a half-wave plate, allows coherence control even when the laser has short temporal coherence length. We generated 14 watts of radially polarized beam from an Ytterbium fiber laser, only limited by the available fiber laser power. PMID- 19550701 TI - Model for polarized and depolarized Rayleigh Brillouin scattering spectra in molecular gases. AB - Numerical models for Rayleigh-Brillouin scattering (RBS) spectra from molecular gases are obtained and discussed in this paper. The current publicly-available S6 model is for polarized RBS spectra only, despite the existence of both polarized and depolarized RBS light in many real applications. One of the new models (Q9) can be used to calculate both polarized and depolarized RBS spectra. In addition, this model has a solid physical ground because it is based on the correct Waldmann-Snider equation in which molecular internal energy is treated quantum mechanically. PMID- 19550702 TI - Spectral dependence of single molecule fluorescence enhancement. AB - The fluorescence from a single molecule can be strongly enhanced near a metal nanoparticle acting as an optical antenna. We demonstrate the spectral tunability of this antenna effect and show that maximum enhancement is achieved when the emission frequency is red-shifted from the surface plasmon resonance of the particle. Our experimental results, using individual gold and silver particles excited at different laser-frequencies, are in good agreement with an analytical theory which predicts a different spectral dependence of the radiative and non radiative decay rates. PMID- 19550703 TI - All-optical format conversions from NRZ to BPSK and QPSK based on nonlinear responses in silicon microring resonators. AB - We propose and numerically verify a novel scheme of all-optical format conversion from non-return-to-zero (NRZ) to binary phase-shift- keying (BPSK) at 160 Gb/s using cascaded microring resonators (CMRR) on a single silicon chip. The conversion is based on large phase shift and flattened intensity-response characteristics in the CMRR. A continuous-wave light experiences different phase shifts controlled by the power of an input NRZ signal with an ~8.8-dB extinction ratio, while maintaining approximately the constant intensity. All-optical format conversion from NRZ to quadrature phase-shift-keying (QPSK) is also demonstrated based on parallel NRZ/BPSK converters in a Mach-Zehnder interferometer structure. PMID- 19550704 TI - Separating polarization components through the electro-optic read-out of photorefractive solitons. AB - Analyzing the propagation dynamics of a light beam of arbitrary linear input polarization in an electro-activated photorefractive soliton we are able to experimentally find the conditions that separate its linear polarization components, mapping them into spatially distinct regions at the crystal output. Extending experiments to the switching scheme based on two oppositely biased solitons, we are able to transform this spatial separation into a separation of two distinct guided modes. The result is a miniaturized electro-optic polarization separator. PMID- 19550705 TI - Localized plasmon-engineered spontaneous emission of CdSe/ZnS nanocrystals closely-packed in the proximity of Ag nanoisland films for controlling emission linewidth, peak, and intensity. AB - Using metallic nanoislands, we demonstrate the localized plasmonic control and modification of the spontaneous emission from closely-packed nanocrystal emitters, leading to significant changes in their collective emission characteristics tuned spectrally and spatially by plasmon coupling. Using randomly-distributed silver nanoislands, we show that the emission linewidth of proximal CdSe/ZnS core-shell quantum dots is reduced by 22% and their peak emission wavelength is shifted by 14nm, while their ensemble photoluminescence is enhanced via radiative energy transfer by 21.6 and 15.1 times compared to the control groups of CdSe/ZnS nanocrystals with identical nano-silver but no dielectric spacer and the same nanocrystals alone, respectively. PMID- 19550706 TI - Refractive index dependence of L3 photonic crystal nano-cavities. AB - We model the optical properties of L3 photonic crystal nano-cavities as a function of the photonic crystal membrane refractive index n using a guided mode expansion method. Band structure calculations revealed that a TE-like full band gap exists for materials of refractive index as low as 1.6. The Q-factor of such cavities showed a super-linear increase with refractive index. By adjusting the relative position of the cavity side holes, the Q-factor was optimised as a function of the photonic crystal membrane refractive index n over the range 1.6 to 3.4. Q-factors in the range 3000-8000 were predicted from absorption free materials in the visible range with refractive index between 2.45 and 2.8. PMID- 19550707 TI - Adjacent crosstalk suppression in a colorless WDM passive optical network. AB - We propose and experimentally demonstrate a novel wavelength-division- multiplexed passive optical network (WDM-PON) scheme for the suppression of adjacent crosstalk arising from the wavelength misalignment in arrayed waveguide gratings (AWGs) between a central office (CO) and a remote node (RN). The adjacent crosstalk suppression is achieved by allocating two different bands to adjacent channels of the AWGs by utilizing interleavers and WDM filters. The transmission performance of the proposed scheme was measured at a 155 Mb/s data stream, and error free transmission with a power penalty less than 0.7 dB was successfully achieved in case of AWG misalignment of 0.3 nm. PMID- 19550708 TI - A fast Gabor wavelet transform for high-precision phase retrieval in spectral interferometry. AB - A fast implementation of the Gabor wavelet transform for phase retrieval in spectral interferometry is discussed. This algorithm is experimentally demonstrated for the characterization of a supercontinuum, using spectral phase interferometry for direct electric-field reconstruction (SPIDER). The performance of wavelet based ridge tracking for frequency demodulation is evaluated and compared to traditional Fourier filtering techniques. It is found that the wavelet based strategy is significantly less susceptible toward experimental noise and does not exhibit cycle slip artifacts. Optimum performance of the Gabor transform is observed for a Heisenberg box with unity aspect ratio. As a result, the phase jitter of 60 individual measurements is reduced by about a factor 2 compared to Fourier filtering, and the detection window increases by 20%. With an optimized implementation, retrieval rates of several 10Hz can be reached, which makes the fast Gabor transform a superior one-to-one replacement even in applications that require video-rate update, such as a real-time SPIDER apparatus. PMID- 19550709 TI - Optofluidic trapping and transport on solid core waveguides within a microfluidic device. AB - In this work we demonstrate an integrated microfluidic/photonic architecture for performing dynamic optofluidic trapping and transport of particles in the evanescent field of solid core waveguides. Our architecture consists of SU-8 polymer waveguides combined with soft lithography defined poly(dimethylsiloxane) (PDMS) microfluidic channels. The forces exerted by the evanescent field result in both the attraction of particles to the waveguide surface and propulsion in the direction of optical propagation both perpendicular and opposite to the direction of pressure-driven flow. Velocities as high as 28 mum/s were achieved for 3 mum diameter polystyrene spheres with an estimated 53.5 mW of guided optical power at the trapping location. The particle-size dependence of the optical forces in such devices is also characterized. PMID- 19550710 TI - Theoretical analysis of supercontinuum generation in a highly birefringent D shaped microstructured optical fiber. AB - This paper carries out a rigorous analysis of supercontinuum generation in an improved highly asymmetric microstructured fiber (MF) design. This geometry, defined simply as D-MF, has the advantage of being produced with a regular stacking and drawing technology. We have obtained birefringence values on the order of 4.87x10(-3) at the adopted pump wavelength and a significantly smaller effective area when compared to a whole MF, which makes this fiber quite attractive for SCG. Therefore, this D-MF design is a promising alternative for SCG since it provides new degrees of freedom to control field confinement, birefringence, and dispersion characteristics of MFs. PMID- 19550711 TI - Moving picture recording and observation of three-dimensional image of femtosecond light pulse propagation. AB - We recorded and observed, for the first time, three-dimensional image of femtosecond light pulse propagation as continuous moving picture using light-in flight recording by holography. We present the moving pictures of collimated and converging light pulses and some images extracted from them. We also discussed inherent feature appearing in the images. Such a discussion is essential to determine the actual shape of the propagating light pulse. This technique provides the means for observation of a temporally and spatially continuous moving picture of light itself and also enables the analysis of various kinds of ultrafast phenomena. PMID- 19550712 TI - Demonstration of a Mid-infrared silicon Raman amplifier. AB - We demonstrate, for the first time, a mid infrared silicon Raman amplifier. Amplification of 12 dB is reported for a signal at 3.39 micron wavelength. The active medium was a 2.5 cm long silicon sample that was pumped with 5ns pulses at 2.88 micron. Such a technology can potentially extend silicon photonics' application beyond data communication in the near IR and into the mid-IR world of remote sensing, biochemical detection and laser medicine. Challenges faced in the mid-IR regime such as a higher free carrier scattering rate longer lifetime in mid-IR waveguides are also discussed. PMID- 19550713 TI - A broad-angle polarization beam splitter based on a simple dielectric periodic structure. AB - We propose a novel broad-angle polarization beam splitter utilizing the spatial dispersion of a special multi-layered dielectric periodic structure. The equal frequency contours of this structure are flat lines for TE polarization but curved lines for TM polarization at a designed frequency. This special multi layered structure has a fixed optical thickness for TE polarization for all incident angles. A polarization beam splitter working over a broad range of angle (from 0 degrees to 70 degrees ) is achieved by stacking two such multi-layered structures of finite length (in the normal direction) with a half-period shift in the transverse direction. PMID- 19550715 TI - Optical liquid ring resonator sensor. AB - We demonstrate a robust and highly responsive optical microsensor, which probes the refractive index of liquids flowing along a ~ 100 mum radius channel formed in a polymer matrix. Sensing is based on measurement of the transmission spectrum of the whispering gallery modes, which are excited across the liquid channel by an optical microfiber imbedded into the polymer. The achieved sensitivity is 800 nm/RIU. Potentially, it is straightforward to assemble the sensing elements of this type into a lab-on-the-chip imbedded in a solidified optical material. PMID- 19550714 TI - Terahertz pulse reflective focal-plane tomography. AB - An effective terahertz (THz) imaging technology is presented for achieving tomographic image. A THz pulse reflective focal-plane imaging system is built up and the tomographic image of a metallic cross hidden by a high resistivity Si wafer is achieved. Using the reflected pulses from each interface, the thickness of each layer can be calculated with calculation error below 2.5%. This work demonstrates that the THz pulse focal-plane tomography can be used to analysis interior configuration of the object. PMID- 19550716 TI - Preform fabrication and fiber drawing of 300 nm broadband Cr-doped fibers. AB - The fabrication of a Cr-doped fiber using a drawing-tower method with Cr:YAG as the core of the preform is presented. The Cr-doped YAG preform was fabricated by a rod-in-tube method. By employing a negative pressure control in drawing-tower technique on the YAG preform, the Cr-doped fibers with a better core circularity and uniformity, and good interface between core and cladding were fabricated. The amplified spontaneous emission spectrum showed a broadband emission of 1.2 to 1.6 mum with the output power density about a few nW/nm. The results indicate that this new Cr-doped fiber may be used as a broadband fiber amplifier to cover the bandwidths in the whole 1.3-1.6 mum range of low-loss and lowdispersion windows of silica fibers. PMID- 19550717 TI - High SBS-threshold, narrowband, erbium codoped with ytterbium fiber amplifier pulses frequency-doubled to 770 nm. AB - We present results of pulsed, narrowband amplification at 1540.6nm using a polarization maintaining, large mode area gain fiber codoped with erbium and ytterbium. At a repetition rate of 55 kHz, 2.9 W of average 1540.6nm power were generated with a pulse duration of 136 ns, corresponding to an SBS free peak power of 360 W. The amplified signal was frequency doubled in peridically poled potassium titanyl phosphate and conversion efficiencies of up to 56% were generated. When varying the repetition rate between 55-150 kHz the conversion efficiency changed from 56% to 35% due to the limited pump power. PMID- 19550718 TI - Spectral gaps and mode localization in Fibonacci chains of metal nanoparticles. AB - In this paper we study the spectral, localization and dispersion properties of dipolar modes in quasi-periodically modulated nanoparticle chains based on the Fibonacci sequence. By developing a transfer matrix approach for the calculation of resonant frequencies, oscillation eigenvectors and integrated density of states (IDS) of spatially-modulated dipole chains, we demonstrate the presence of large spectral gaps and calculate the pseudo-dispersion diagram of Fibonacci plasmonic chains. The presence of plasmonic band-gaps and localized states in metal nanoparticle chains based on quasi-periodic order can have a large impact in the design and fabrication of novel nanophotonics devices. PMID- 19550719 TI - 3.4-mum ZGP RISTRA nanosecond optical parametric oscillator pumped by a 2.05-mum Ho:YLF MOPA system. AB - We report on the first demonstration of ZGP OPO based on Rotated Image Singly Resonant Twisted RectAngle (RISTRA) cavity. For the OPO signal wave we achieved a near diffraction-limited beam at 3.4 mum with pulse energy of 10 mJ at repetition rate up to 500 Hz. As a pump source for the ZGP OPO, we utilized a 2-mum, TEM00, Ho:YLF MOPA system producing > 55 mJ energy per pulse at repetition rate range from single shot to 500 Hz. PMID- 19550720 TI - Long, low loss etched As(2)S(3) chalcogenide waveguides for all-optical signal regeneration. AB - We report on the fabrication and optical properties of etched highly nonlinear As(2)S(3) chalcogenide planar rib waveguides with lengths up to 22.5 cm and optical losses as low as 0.05 dB/cm at 1550 nm - the lowest ever reported. We demonstrate strong spectral broadening of 1.2 ps pulses, in good agreement with simulations, and find that the ratio of nonlinearity and dispersion linearizes the pulse chirp, reducing the spectral oscillations caused by self-phase modulation alone. When combined with a spectrally offset band-pass filter, this gives rise to a nonlinear transfer function suitable for all-optical regeneration of high data rate signals. PMID- 19550721 TI - Study of color-conversion-materials in chromatic-stability white organic light emitting diodes. AB - We have fabricated high efficiency white organic light-emitting diodes (WOLEDs) with red-dye doped hole injection layer (HIL) and blue emissive layer. We investigated the carrier transport and photoluminescence efficiency in the device of single-layer HIL with different red dye species, bis(4-diphneylamino-phenyl) perylene-3-yl-amine, 4-(dicyanomethylene)-2-t-butyl-6-(1,1,7,7 tetramethyljulolidyl-9enyl)-4Hpyran and a dopant material of fused aromatic ring (P1) provided by Idemitsu Kosan in 4,4',4"-tris[2 naphthyl(phenyl)amino]triphenylamine with a varied concentration. It was found that 25-% wt P1 in HIL has better electro-optical characteristics. Furthermore, we have fabricated three WOLEDs of chromatic-stability with different thickness of HIL layer. High chromatic-stability CCM-WOLEDs have been achieved, which can be applied for lightening and backlight for flat panel display. PMID- 19550722 TI - Efficient and spurious-free integral-equation-based optical waveguide mode solver. AB - Modal analysis of waveguides and resonators by integra-lequation formulations can be hindered by the existence of spurious solutions. In this paper, spurious solutions are shown to be eliminated by introduction of a Rayleigh-quotient based matrix singularity measure. Once the spurious solutions are eliminated, the true modes may be determined efficiently and reliably, even in the presence of degeneracy, by an adaptive search algorithm. Analysis examples that demonstrate the efficacy of the method include an elliptical dielectric waveguide, two unequal touching dielectric cylinders, a plasmonic waveguide, and a realistic micro-structured optical fiber. A freely downloadable version of an optical waveguide mode solver based on this article is available. PMID- 19550723 TI - Computing Photonic Crystal Defect Modes by Dirichlet-to-Neumann Maps. AB - We develop an efficient numerical method for computing defect modes in two dimensional photonic crystals based on the Dirichletto- Neumann (DtN) maps of the defect and normal unit cells. The DtN map of a unit cell is an operator that maps the wave field on the boundary of the cell to its normal derivative. The frequencies of the defect modes are solved from a condition that a small matrix is singular. The size of the matrix is related to the number of points used to discretize the boundary of the defect cell. The matrix is obtained by solving a linear system involving only discrete points on the edges of the unit cells in a truncated domain. PMID- 19550724 TI - Compact silicon microring resonators with ultra-low propagation loss in the C band. AB - The propagation loss in compact silicon microring resonators is optimized with varied ring widths as well as bending radii. At the telecom band of 1.53-1.57 mum, we demonstrate as low as 3-4 dB/cm propagation losses in compact silicon microring resonators with a small bending radius of 5 mum, corresponding to a high intrinsic quality factor of 200,000-300,000. The loss is reduced to 2-3 dB/cm for a larger bending radius of 10 mum, and the intrinsic quality factor increases up to an ultrahigh value of 420,000. Slot-waveguide microring resonators with around 80% optical power confinement in the slot are also demonstrated with propagation losses as low as 1.3+/-0.2 dB/mm at 1.55 mum band. These loss numbers are believed to be among the lowest ones ever achieved in silicon microring resonators with similar sizes. PMID- 19550725 TI - 4W continuous-wave narrow-linewidth tunable solid-state laser source at 546nm by externally frequency doubling a ytterbium-doped single-mode fiber laser system. AB - A high-power continuous-wave coherent light source at 545.5nm is described. We use 8.3W from a solid-state ytterbium-doped single-mode fiber oscillator/amplifier system as input into an external frequency doubling stage. This system produces up to 4.1W of stable green single-frequency laser radiation. We characterize the light source by performing absorption spectroscopy on iodine across the full tuning range of the fiber laser and saturation spectroscopy on one strong iodine line of the doppler-broadened spectrum. PMID- 19550726 TI - Portable, large-bandwidth time-resolved system for diffuse optical spectroscopy. AB - We report the development of a compact time-resolved system for the measurement of the optical properties of highly scattering media over a bandwidth of 600-1000 nm. The instrument is based on a fiber laser generating supercontinuum radiation, that is spectrally dispersed and sequentially used to illuminate the sample. A single photon avalanche photo-diode in combination with time correlated single photon counting is used to recover the time-dispersion curve at each wavelength, both fitted by the diffusion equation. Transmittance measurements performed on calibrated epoxy phantoms and in-vivo on female breast are presented, showing good agreement with previous reports. PMID- 19550727 TI - Parallel photopolymerisation with complex light patterns generated by diffractive optical elements. AB - Photopolymerisation by scanning a focused laser beam is a powerful method to build structures of arbitrary complexity with submicrometer resolution. We introduce parallel photopolymerisation to enhance the efficiency. Instead of multidimensional scanning of a single focus, the structure is generated simultaneously with diffractive patterns. We used fixed diffractive optical elements (DOEs), kinoforms, and Spatial Light Modulators (SLMs). The possibilities of photopolymerisation using SLM were investigated: the added flexibility using the programmable device is demonstrated. By using these DOEs, straight and helical cross shaped columns were produced with a single scan at a rate about an order of magnitude faster than by simple scanning. The produced helical structures could be rotated by optical tweezers. PMID- 19550728 TI - Defect superlattice solitons. AB - We reveal theoretically that defect superlattice solitons (DSSs) exist at the defect site in one-dimensional optical superlattices with focusing saturable nonlinearity. Solitons with some unique properties exist in superlattices with defects. For a positive defect, solitons exist at the semi-infinite gap, and solitons are stable at low power but unstable at high power. For a negative defect, most solitons exist in the first finite gap and can propagate stably. In particular, it is found that the solitons can be divided into two equal parts upon propagation in a certain regime of parameters. PMID- 19550729 TI - Elevation information in tail (EIT) technique for lidar altimetry. AB - A technique we refer to as Elevation Information in Tail (EIT) has been developed to provide improved lidar altimetry from CALIPSO lidar data. The EIT technique is demonstrated using CALIPSO data and is applicable to other similar lidar systems with low-pass filters. The technique relies on an observed relation between the shape of the surface return signals (peak shape) and the detector photo multiplier tube transient response (transient response tail). Application of the EIT to CALIPSO data resulted in an order of magnitude or better improvement in the CALIPSO land surface 30-meter elevation measurements. The results of EIT compared very well with the National Elevation Database (NED) high resolution elevation maps, and with the elevation measurements from the Shuttle Radar Topography Mission (SRTM). PMID- 19550730 TI - Composite Yb:YAG/Cr(4+):YAG ceramics picosecond microchip lasers. AB - Efficient laser-diode pumped picosecond self-Q-switched all-ceramic composite Yb:YAG/Cr(4+):YAG microchip lasers with 0.72 MW peak power has been developed. Lasers with nearly diffraction-limited beam quality (M(2) < 1.09), oscillate at stable single- and multi- longitudinal-modes due to the combined etalon effects in the Yb:YAG and Cr(4+):YAG parts of its binary structure. PMID- 19550731 TI - Orientation-controllable self-organized microgratings induced in the bulk SrTiO(3) crystal by a single femtosecond laser beam. AB - Self-organized microgratings were induced in the bulk SrTiO(3) crystal by readily scanning the laser focus in the direction perpendicular to the laser propagation axis. The groove orientations of those gratings could be controlled by changing the irradiation pulse number per unit scanning length, which could be implemented either through adjusting the scanning velocity at a fixed pulse repetition rate or through varying the pulse repetition rate at a fixed scanning velocity. This high-speed method for fabrication of microgratings will have many potential applications in the integration of micro-optical elements. The possible formation mechanism of the self-organized microgratings is also discussed. PMID- 19550732 TI - A mobile head-worn projection display. AB - A recent advancement was achieved in the integration and miniaturization of a binocular head-worn projection display (HWPD) conceived for fully mobile users. The devised display, referred to as Mobile HWPD (M-HWPD), offers see-through capability through custom-designed, light-weight projection optics and an integrated commercial-off-the-shelf (COTS) retro-reflective screen to display full color stereoscopic rendered images augmenting the real world. Moreover, the light-weight optical device (i.e., approximately 8g per eye) has the ability to project clear images at three different locations within near- or far-field observation depths without loss of image quality. In this paper, we first demonstrate the miniaturization of the optics, the optical performance, and the integration of these components with the retro-reflective screen to produce an M HWPD prototype. We then show results that demonstrate the feasibility of superimposing computer-generated images on a real outdoor scene with the M-HWPD. PMID- 19550733 TI - SiPM for fast Photon-Counting and Multiphoton Detection. AB - We demonstrate fast counting and multiphoton detection abilities of a Silicon Photo Multiplier (SiPM). In fast counting mode we are able to detect two consecutive photons separated by only 2.3 ns corresponding to 430 MHz. The counting efficiency for small optical intensities at lambda= 532 nm was found to be around 16% with a dark count rate of 52 kHz at T= -5 masculine C. Using the SiPM in multiphoton detection mode, we find a good signal discrimination for different numbers of simultaneously detected photons. PMID- 19550734 TI - Facile fabrication of close-packed microlens arrays using photoinduced surface relief structures as templates. AB - We demonstrate the cost-effective and facile method of fabricating close-packed microlens arrays using photoinduced two-dimensional (2-D) surface relief structures as original templates. 2-D surface relief structures are produced by successive inscription of two beams interference patterns with different grating vectors on azopolymer films. The employed exposure dose of 1st inscription stage and 2nd inscription stage are optimized to obtain symmetrical modulation heights. These photoinduced 2-D surface relief structures on azopolymer films are used directly to mold PDMS, and PDMS molds were then transferred onto photopolymer to imprint microlens arrays. Using this method, tetragonally and hexagonally close packed microlens arrays are successfully fabricated in rapid and cost-effective way. PMID- 19550735 TI - Bouncing of a dispersive pulse on an accelerating soliton and stepwise frequency conversion in optical fibers. AB - We demonstrate that a short pulse with spectrum in the range of normal group velocity dispersion can experience periodic reflections on a refractive index maximum created by a co-propagating with it soliton, providing the latter is continuously decelerated by the intrapulse Raman scattering. After each reflection the intensity profile and phase of the pulse are almost perfectly reconstructed, while its frequency is stepwise converted. This phenomenon has direct analogy with the effect of 'quantum bouncing' known for cold atoms. PMID- 19550736 TI - Low-loss high-index-contrast planar waveguides with graded-index cladding layers. AB - We experimentally demonstrate, for the first time, propagation loss reduction via graded-index (GRIN) cladding layers in high-index-contrast (HIC) glass waveguides. We show that scattering loss arising from sidewall roughness can be significantly reduced without compromising the high-index-contrast condition, by inserting thin GRIN cladding layers with refractive indices intermediate between the core and topmost cover of a strip waveguide. Loss as low as 1.5 dB/cm is achieved in small core (1.6 mum x 0.35 mum), high-index-contrast (Deltan = 1.37) arsenic-based sulfide strip waveguides. This GRIN cladding design is generally applicable to HIC waveguide systems such as Si/SiO2. PMID- 19550737 TI - Microlens diffusers for efficient laser speckle generation. AB - Laser Speckle is the optical phenomena resulting from the random interference of coherent light. This phenomenon can be utilized to measure the Modulation Transfer Function (MTF) of detector arrays. Common devices used for speckle generation, such as integrating spheres and ground glass, suffer from low efficiencies less than 20%. Microlens diffusers are shown to be more efficient alternatives for speckle generation. An analysis of the statistical behavior of microlens diffusers is presented with emphasis on their application to MTF testing of detector arrays in the visible spectrum. PMID- 19550738 TI - Yb(3+) spectroscopy in (Nb or Ta):RbTiOPO(4) single crystals for laser applications. AB - Single crystals of Yb:RbTiOPO(4) codoped with Nb(5+) or Ta(5+) were grown by the top seeded solution growth slow-cooling technique. The ytterbium concentration in the crystals varies as a function of the molar ratio of the precursor oxides and of the codopant, reaching a maximum value of 1.9x10(20) Yb(3+) ions/cm(3). The broad band near 1 mum in absorption and emission spectra at room temperature is due to the large splitting of the Yb(3+) ground state. The ytterbium (2)F(5/2) level radiative lifetime in Nb:RbTiOPO(4) (tau(rad) = 2.7 ms), was calculated and then compared to the measured fluorescence decay time (tau(em) = 2.2 ms), giving an intrinsic quantum efficiency of 81%. To evaluate the potentiality of these crystals for self-frequency doubling, preliminary results of Yb(3+) laser operation and fundamental wavelength measurements for type-II non-critical second harmonic generation ( lambda(NCPM)) are also reported. PMID- 19550739 TI - Quantitative Phase Microscopy of microstructures with extended measurement range and correction of chromatic aberrations by multiwavelength digital holography. AB - Quantitative Phase Microscopy (QPM) by interferometric techniques can require a multiwavelength configuration to remove 2pi ambiguity and improve accuracy. However, severe chromatic aberration can affect the resulting phase-contrast map. By means of classical interference microscope configuration it is quite unpractical to correct such aberration. We propose and demonstrate that by Digital Holography (DH) in a microscope configuration it is possible to clear out the QPM map from the chromatic aberration in a simpler and more effective way with respect to other approaches. The proposed method takes benefit of the unique feature of DH to record in a plane out-of-focus and subsequently reconstruct numerically at the right focal image plane. In fact, the main effect of the chromatic aberration is to shift differently the correct focal image plane at each wavelength and this can be readily compensated by adjusting the corresponding reconstruction distance for each wavelength. A procedure is described in order to determine easily the relative focal shift among different imaging wavelengths by performing a scanning of the numerical reconstruction along the optical axis, to find out the focus and to remove at the same time the chromatic aberration. PMID- 19550740 TI - An optoelectronic measurement system for measuring 6-degree-of-freedom motion error of rotary parts. AB - This paper presents an optoelectronic measurement system for measuring 6 degree of-freedom (DOF) motion error of rotary parts. It comprises a pyramid-polygon mirror, three laser diodes and three 2-axis position sensing detectors (PSD). The laser/PSD pairs are arranged evenly around the pyramid-polygon-mirror, which is mounted rigidly on and aligned axially with the rotary part to be measured. Laser rays from the laser diodes are reflected off the respective mirrors to the respective PSDs. The incidence point of the laser ray on the PSD's surface varies with the pose of the pyramid-polygon-mirror, allowing the PSD to register variation in the mirror and, thereby, the rotary part. With appropriate orientation of the lasers and PSDs, this system can measure variation (error) during rotation of a rotary part. By use of skew-ray tracing and first order Taylor series expansion, the system achieves measurement of translational and rotational motion errors for each Cartesian axis. To validate the proposed methodology, a laboratory prototype system is built. System verification and stability tests are conducted to evaluate its performance. Stability test results show that measurement errors and maximum crosstalk are within +/-1 mum in translation and +/-1.5 arc sec in rotation. PMID- 19550741 TI - Optical trapping of nonspherical particles in the T-matrix formalism: erratum. AB - An erratum is presented to correct the errors in two equations in Sect. 2 of [Opt. Express 15, 11984-11998 (2007)]. PMID- 19550742 TI - Dual-probe near-field fiber head with gap servo control for data storage applications. AB - We present a novel fiber-based near-field optical head consisting of a straw shaped writing probe and a flat gap sensing probe. The straw-shaped probe with a C-aperture on the end face exhibits enhanced transmission by a factor of 3 orders of magnitude over a conventional fiber probe due to a hybrid effect that excites both propagation modes and surface plasmon waves. In the gap sensing probe, the spacing between the probe and the disk surface functions as an external cavity. The high sensitivity of the output power to the change in the gap width is used as a feedback control signal. We characterize and design the straw-shaped writing probe and the flat gap sensing probe. The dual-probe system is installed on a conventional biaxial actuator to demonstrate the capability of flying over a disk surface with nanometer position precision. PMID- 19550743 TI - Tunable transmission at 100 THz through a metallic hole array with a varying hole channel shape. AB - Extraordinary optical transmission spectrum for a two-dimensional metallic hole array (2D-MHA) changes with the hole channel shape. In this paper a new converging-diverging channel (CDC) shape is proposed. A three-dimensional (3D) finite element method is utilized to analyze the transmission characteristics of the 2D-MHA with CDC. The transmission peaks are blue-shifted when the gap at the throat of CDC is reduced. Similar blue-shift in the transmission peaks are observed for a straight channel MHA when the aperture size is reduced. The transmission for the straight channel MHA is not sensitive to the metal film thickness. But, for a CDC MHA the transmission varies with the metal film thickness. Also, the CDC shape gives an extra degree of geometrical variable to 2D-MHA for tuning the transmission peak location with potential applications in nanolithography, imaging and biosensing. PMID- 19550744 TI - Azimuthal distinguishability of entangled photons generated in spontaneous parametric down-conversion. AB - We experimentally demonstrate that paired photons generated in different sections of a down-conversion cone, when some of the interacting waves show Poynting vector walk-off, carry different spatial correlations, and therefore a different degree of spatial entanglement. This is shown to be in agreement with theoretical results. We also discuss how this azimuthal distinguishing information of the down-conversion cone is relevant for the implementation of quantum sources aimed at the generation of entanglement in other degrees of freedom, such as polarization. PMID- 19550745 TI - High-efficiency and high-contrast phosphorescent top-emitting organic light emitting devices with p-type Si anodes. AB - We report high-efficiency and high-contrast phosphorescent topemitting organic light-emitting devices (OLEDs) by employing the low reflectance p-type Si bottom anodes and the high transmittance Cs(2)CO(3)/Ag top cathodes for effective hole and electron injection. With the green electrophosphorescent material fac tris (2 phenylpyridine) iridium [Ir(ppy)(3)] doped emitting layer, the devices exhibit peak external quantum and power efficiencies of 3.5% (12 cd/A) and 4.5 lm/W, which are the highest values reported for OLEDs using Si wafers as electrodes. Moreover, these devices exhibit significantly higher contrast compared to the conventional bottom-emitting and top-emitting OLEDs with the highly reflective back electrodes. PMID- 19550746 TI - Reciprocal transmissions and asymmetric modal distributions in waveguide-coupled spiral-shaped microdisk resonators. AB - We report transmission spectra and mode-field distributions of a waveguide coupled spiral-shaped microdisk resonator on a silicon nitride-on-silica substrate. Our measured and simulated transmission spectra reveal reciprocal transmissions between clockwise and counterclockwise travelingwaves of such microcavity that lacks mirror symmetry. Our measured out-of-plane scattering intensity distributions and simulated steady-state mode-field patterns, however, indicate asymmetric modal distributions that depend on the sense of lightwave circulations and the input-coupling mechanisms. We discuss implications of the observed reciprocal transmissions with asymmetric modal distributions to unidirectional lasing from spiral-shaped microcavities reported in the literature. PMID- 19550747 TI - Switchable optical UWB monocycle and doublet generation using a reconfigurable photonic microwave delay-line filter. AB - Optically switchable Ultra-Wideband (UWB) monocycle and doublet pulse generation using an optically reconfigurable photonic microwave delay-line filter is proposed and demonstrated. The microwave filter can be reconfigured as a two- or three-tap microwave filter with coefficients of (1, -1) or (1, -2, 1). The function of the two- or three-tap microwave filter is equivalent to an operation of a first- or second-order difference, which can be approximated as a first- or second-order derivative. When a Gaussian pulse is inputted to the two- or three tap microwave delay-line filter, a Gaussian monocycle or doublet pulse is generated. The proposed photonic microwave delay-line filter is implemented using a polarization modulator (PolM), a length of polarization maintaining fiber (PMF), and a balanced photo-detector (BPD). In the experiment, Gaussian monocycle and doublet pulses with a fractional bandwidth of about 170% and 130% are generated. The switchability of the proposed UWB pulse generator in pulse shape and polarity is also experimentally demonstrated. PMID- 19550748 TI - Reflection and emission properties of an infrared emitter. AB - The reflection and emission properties of an infrared emitter, which is a plasmonic multilayer structure consisting of a relief metallic grating, a waveguide layer, and a metallic substrate are investigated both experimentally and theoretically. A localized surface plasmon polariton (SPP) mode which is angular-independent in almost the full range of incident angles is observed. The thermal emission of this structure is also measured. It is found that the emission peak coincides with the angular-independent localized SPP mode. In addition, the emission spectrum of the plasmonic emitter can be predicted by investigating the reflectance spectrum. PMID- 19550749 TI - Optically pumped Si nanocrystal emitter integrated with low loss silicon nitride waveguides. AB - We describe the integration of optically pumped silicon nanocrystals (Si-ncs) embedded in SiO(2) with low loss silicon nitride slab waveguides. An emission waveguide containing Si-ncs with a broad band emission centered at 850 nm, together with a low loss transmission silicon nitride waveguide forms a two section device. The waveguides are fabricated via the deposition of SiO(x) and silicon nitride using ECR-PECVD. Incorporation of hydrogen through annealing, while beneficial to emission from the Si-ncs, is found to increase material absorption in silicon nitride. This is reconciled by annealing at low temperature. This work shows clearly the potential for this material system as a means for the integration of optical emission and waveguiding using a wholly VLSI compatible processing technology. We further suggest that immediate applications exist in particular in the field of evanescent sensing. PMID- 19550750 TI - Theory of light emission from a dipole source embedded in a chiral sculptured thin film. AB - Developing a theory based on a spectral Green function for light emission from a point-dipole source embedded in a chiral sculptured thin film (CSTF), we found that the intensity and polarization of the emitted light are strongly influenced by the structural handedness of the CSTF as well as the placement and orientation of the source dipole. The emission patterns across both pupils of the dipole containing CSTF can be explained in terms of the circular Bragg phenomenon exhibited by CSTFs when illuminated by normally as well as obliquely incident plane waves. The emission characteristics augur well for the future of CSTFs as optical biosensors as well as light emitters with controlled circular polarization and bandwidth. PMID- 19550751 TI - Period-doubling and total mode-locking, and chaotic pulsations in a GaAs ridge wave-guide external-cavity diode laser. AB - We report on the experimental observations of a period-doubling route to chaos and a total mode-locking between two lowest-order lateral modes of a GaAs ridge wave-guide diode laser at 795 nm. A self-starting passively mode-locking was achieved in an extended-cavity diode laser (ECDL) operating in a gain saturation regime. In the period-doubling mode-locked state, the RF spectra associated with the coherently coupled lateral TE(0) and TE(1) modes, consisted of harmonics (nf(rep)/2, n is an integer) of the half of pulse repetition rate f(rep)/2 and the period (2T) of the pulse train was two-times longer than the pulse round-trip time T. On the other hand, in the total mode-locked state, the RF spectra and the pulse train corresponding to the TE(0) and TE(1) modes had exactly the same features compared to those of the period-doubling mode-locked state, except the RF spectra and the pulse train of the TE(1) mode were shifted by f(ref)/4 in frequency and by T in time, respectively, indicating the pulses actually traveled alternatively through two lateral modes. The total mode-locking and also chaotic pulsations were observed at slightly different operation parameters, e.g. at different feedback angle of the grating, which was used as an output coupler of the ECDL. PMID- 19550752 TI - Electric field enhancement between two Si microdisks. AB - The field enhancement in the gap between two Si microdisks is theoretically investigated using the finite difference time domain method. We show that the electric field within this gap increases as the distance between the two disks decreases, and it can be enhanced by as much as two orders of magnitude. By perturbing the Si microdisks to force the field leakage into an ever smaller volume, the field enhancement can reach a value as high as 238 with a deep sub wavelength mode volume. This behavior is comparable to what can be observed in gap plasmons between metal nanoparticles, but is produced here in purely dielectric structures. PMID- 19550753 TI - Investigation of aqueous alcohol and sugar solutions with reflection terahertz time-domain spectroscopy. AB - We give a detailed analysis of a general realization of reflection terahertz time domain spectroscopy. The method is self-referenced and applicable at all incidence angles and for all polarizations of the incident terahertz radiation. Hence it is a general method for the determination of the dielectric properties of especially liquids in environments where transmission measurements are difficult. We investigate the dielectric properties in the 0.1-1.0 THz frequency range of liquids using reflection terahertz time-domain spectroscopy. We apply the technique for the determination of alcohol and sugar concentration of commercial alcoholic beverages and liquors. The special geometry of the experiment allows measurement on sparkling beverages. PMID- 19550754 TI - An assessment of the Wigner distribution method in Doppler OCT. AB - Analyzing the experimental data of the velocity distribution in a fluid flow using Doppler Optical Coherence Tomography (OCT), we compared the Wigner distribution method to the short-time Fourier transform method, the Hilbert-based phase-resolved method and the autocorrelation method. We conclude that the pseudo Wigner-distribution signal processing method is overall more precise than other often-used methods in Doppler OCT for the analysis of cross-sectional velocity distributions. PMID- 19550755 TI - Sub-critical regime of femtosecond inscription. AB - We apply well known nonlinear diffraction theory governing focusing of a powerful light beam of arbitrary shape in medium with Kerr nonlinearity to the analysis of femtosecond (fs) laser processing of dielectric in sub-critical (input power less than the critical power of self-focusing) regime. Simple analytical expressions are derived for the input beam power and spatial focusing parameter (numerical aperture) that are required for achieving an inscription threshold. Application of non-Gaussian laser beams for better controlled fs inscription at higher powers is also discussed. PMID- 19550756 TI - A highly compact third-order silicon microring add-drop filter with a very large free spectral range, a flat passband and a low delay dispersion. AB - We demonstrate highly compact third-order silicon microring add-drop filters. The microring resonator has a small radius of 2.5 mum and a very large free spectral range of 32 nm at 1.55 mum. Experimental results show a low add-drop crosstalk of around -20 dB. Box-like channel dropping response is demonstrated, and it has a passband of ~ 1 nm (125 GHz), fast rolling-off (slope ~ 0.2 dB/GHz), high out-of band signal rejection of around 40 dB and a low drop loss. Simulation agrees well with experiments in power transmission, and the group delay is also simulated and the variation is less than 1 ps within the passband. The propagation loss in microring resonators is optimized. PMID- 19550757 TI - Transformation-designed optical elements. AB - We describe transformation design of optical elements which, in addition to image transfer, perform useful operations. For one class of operations, including translation, rotation, mirroring and inversion, an image can be generated that is ideal in the sense of the perfect lens (combining both near- and far-field components in a flat, unit transfer function, up to the limits imposed by material imperfection). We also describe elements that perform magnification, free from geometric aberrations, even while providing free-space working distance on both the input and output sides. These magnifying elements also operate in the near- and far-field, allowing them to transfer near field information into the far field, as with the hyper lens and other related devices, however in contrast to those devices, insertion loss can be much lower, due to the matching properties accessible with transformation design. The devices here described inherently require dispersive materials, thus chromatic aberration will be present, and the bandwidth limited. PMID- 19550758 TI - Design of a LiNbO(3) ribbon waveguide for efficient difference-frequency generation of terahertz wave in the collinear configuration. AB - We propose and investigate a ribbon waveguide for difference-frequency generation of terahertz (THz) wave from infrared light sources. The proposed ribbon waveguide is composed of a nonlinear optic crystal and has a thickness less than the wavelength of the THz wave to support the surface-wave mode in the THz region. By utilizing the waveguide dispersion of the surface-wave mode, the phase matching condition between infrared pump, idler and THz waves can be realized in the collinear configuration. Owing to the weak mode confinement of the THz wave, the absorption coefficient can also be reduced. We design the ribbon waveguide which uses LiNbO(3) crystal and discuss the phase-matching condition for DFG of THz wave. Highly efficient THz-wave generation is confirmed by numerical simulations. PMID- 19550759 TI - Using colloidal lithography to fabricate and optimize sub-wavelength pyramidal and honeycomb structures in solar cells. AB - The external quantum efficiency of solar cells can be improved by using texturing pyramid- and honeycomb-like structures with minimum reflection. In this study, we investigated the reflection properties of texturing structures through rigorous coupled-wave analysis and the three-dimensional finite-difference time domains (FDTD) method to analyze close-packed texturing structures. We also demonstrate a simple method-combining sub-wavelength-scale monolayer and bilayer polystyrene spheres with a one-step reactive ion etching process-to fabricate optimized pyramid- and honeycomb-shaped antireflection structures, respectively. Thus, sub wavelength pyramidal and honeycomb-like structures displaying low reflectance were obtained readily without the need for any lithography equipment. PMID- 19550760 TI - Highly sensitive surface plasmon terahertz imaging with planar plasmonic crystals. AB - We report on the operation of a highly sensitive terahertz imaging system relying on a planar metallic plasmonic crystal as a terahertz surface plasmon resonant (THz-SPR) sensor. The terahertz imaging is based on the resonantly enhanced transmission phenomenon of a periodically perforated metal film. The detection sensitivity and the imaging contrast for small amounts of substance are considerably better than those of the conventional terahertz transmission imaging without a THz-SPR sensor. As a demonstration, a high contrast image of a fingerprint recorded on a thin film can be achieved by using this system. PMID- 19550761 TI - Novel cascaded injection-locked 1.55-mum VCSELs with 66 GHz modulation bandwidth. AB - We demonstrate a novel cascaded configuration of optically injection-locked (COIL) VCSELs, which enables a wide and tailorable direct modulation bandwidth. Up to 66 GHz bandwidth is achieved using VCSELs with an original, free-running 10 GHz bandwidth. Different configurations of cascading are discussed in detail with the focus on optimizing the modulation bandwidth. We also discuss scaling capability of this technique to achieve tailorable modulation response. PMID- 19550762 TI - Joint nonuniform illumination estimation and deblurring for bar code signals. AB - We present a novel joint nonuniform illumination estimation and deblurring method for bar code signals based on a penalized nonlinear squares objective function. The objective function is based on the proper parameterization of a bar code signal and nonuniform illumination as well as a regularization on the illumination using a smoothness penalty. By the minimization of the objective function, the proposed method simultaneously estimates the bar code signal and illumination in the spatial domain. In simulations and experiments, the proposed method showed improved performance compared with two conventional bar code decoding methods without deblurring or nonuniform illumination correction. In a few iterations, the proposed method was able to decode test bar code signals that were not decodable due to blurring or nonuniform illumination. PMID- 19550763 TI - Photodarkening in Yb doped fibers: experimental evidence of equilibrium states depending on the pump power. AB - Photodarkening in Yb doped fibers was examined at 633 nm in-situ during cladding pumping at 915 nm with varying pump powers and with no indication of an onset threshold. For the first time, the partial bleaching of the photodarkening loss by the pump power itself was observed. We found the relaxation to well-defined equilibrium states of the core excess loss, depending on the Yb inversion. From the dependence of the measured rate constant on the density of excited Yb ions we conclude, that on average 3 to 4 excited Yb ions create or bleach one color center responsible for the core excess loss. PMID- 19550764 TI - Improved detection of atmospheric turbulence with SLODAR. AB - We discuss several improvements in the detection of atmospheric turbulence using SLOpe Detection And Ranging (SLODAR). Frequently, SLODAR observations have shown strong ground-layer turbulence, which is beneficial to adaptive optics. We show that current methods which neglect atmospheric propagation effects can underestimate the strength of high altitude turbulence by up to ~ 30%. We show that mirror and dome seeing turbulence can be a significant fraction of measured ground-layer turbulence, some cases up to ~ 50%. We also demonstrate a novel technique to improve the nominal height resolution, by a factor of 3, called Generalized SLODAR. This can be applied when sampling high-altitude turbulence, where the nominal height resolution is the poorest, or for resolving details in the important ground-layer. PMID- 19550765 TI - Demonstration of air-guided quantum cascade lasers without top claddings. AB - We report on quantum cascade lasers employing waveguides based on a predominant air confinement mechanism in which the active region is located immediately at the device top surface. The lasers employ ridge-waveguide resonators with narrow lateral electrical contacts only, with a large, central top region not covered by metallization layers. Devices based on this principle have been reported in the past; however, they employed a thick, doped top-cladding layer in order to allow for uniform current injection. We find that the in-plane conductivity of the active region - when the material used is of high quality - provides adequate electrical injection. As a consequence, the devices demonstrated in this work are thinner, and most importantly they can simultaneously support air-guided and surface-plasmon waveguide modes. When the lateral contacts are narrow, the optical mode is mostly located below the air-semiconductor interface. The mode is predominantly air-guided and it leaks from the top surface into the surrounding environment, suggesting that these lasers could be employed for surface-sensing applications. These laser modes are found to operate up to room temperature under pulsed injection, with an emission spectrum centered around l (1/4) 7:66 mum. PMID- 19550766 TI - Photon pair-state preparation with tailored spectral properties by spontaneous four-wave mixing in photonic-crystal fiber. AB - We study theoretically the generation of photon pairs by spontaneous four-wave mixing (SFWM) in photonic crystal optical fiber. We show that it is possible to engineer two-photon states with specific spectral correlation ("entanglement") properties suitable for quantum information processing applications. We focus on the case exhibiting no spectral correlations in the two-photon component of the state, which we call factorability, and which allows heralding of single-photon pure-state wave packets without the need for spectral post filtering. We show that spontaneous four wave mixing exhibits a remarkable flexibility, permitting a wider class of two-photon states, including ultra-broadband, highly anticorrelated states. PMID- 19550767 TI - Frequency-dependent electric dc power consumption model including quantum conversion efficiencies in ultrafast all-optical semiconductor gates around 160 Gb/s. AB - Based on nine up-to-date types of semiconductor-optical-amplifier (SOA) samples, we devised a power-consumption model of SOA-based all-optical gates as a tool to develop faster and more efficient OTDM systems for bitrates from 10 to 160 Gb/s and those over 160 Gb/s. The conventional effect of a continuous wave (cw) holding beam was included in the model. Furthermore, in this work we defined three step-wise quantum conversion efficiencies eta(1), eta(2), and eta(3) from current-injected carriers through photons. The dependence of each of the three efficiencies on the SOA-structure was studied. The total efficiency eta(T) observed for the nine SOAs ranged widely from 0.07 to 0.46. The validity of the power-consumption model was verified by systematically measuring the effective carrier recovery rate. According to our model, the power consumption of the best existing SOA-based gate for 160-Gb/s signals is 750 mW, and this increases at a rate approximately proportional to (bitrate)(2), and decreases proportionally to (1/etaT)(2). PMID- 19550768 TI - Generation of two-mode optical signals with broadband frequency tunability and low spurious signal level. AB - For continuous millimeter and terahertz-wave applications, a two-mode optical signal generation technique that uses two arrayed waveguide gratings and two optical switch units is presented. In addition to easy and fast operation, this scheme offers broadband frequency tunability and high signal purity with a low spurious mode level. Mode spacing, which corresponds to the frequency of the generated MM/THz-wave signal after photomixing, was successfully swept in the range of 200 ~ 550 GHz and the optical spurious mode suppression ratio higher than 25 dBc was achieved. In addition, spurious modes characteristics were investigated by using second harmonic generation (SHG) autocorrelation methods for several frequencies. PMID- 19550769 TI - Study of an efficient longitudinal multimode pumping scheme for Si-nc sensitized EDWAs. AB - We present an efficient multimode longitudinal pumping scheme which overcomes the main limitations of single-mode longitudinal pumping as well as top pumping in Si nanoclusters sensitized Erbium-doped waveguide amplifiers. The proposed configuration is based on evanescent pump light coupling from a multimode waveguide to a Si-nanoclusters sensitized Er(3+)-doped active core. Theoretical predictions, based on propagation and population-rate equations for the coupled Er(3+)/Sinanoclusters system, point out that the proposed pumping scheme can provide high pump intensity within the active core, also ensuring good uniformity of the population inversion along the waveguide amplifier. Although longitudinal multimode pumping by high power LEDs in the visible can potentially lead to low cost integrated amplifiers, further material optimization is required. In particular, we show that when dealing with high pump intensities, confined carrier absorption seriously affects the amplifier performance, and an optimization of both Si-nc and Er(3+) concentrations is necessary. PMID- 19550770 TI - Nanometric control of the distance between plasmonic nanoparticles using optical forces. AB - We theoretically analyze the optical forces between two nearby silver nanoparticles for the case when the wavelength of the incoming light is close to the localized surface plasmon resonance (LSPR). It is shown that the optical force between the nanoparticles is enhanced by the LSPR and that it changes from attractive to repulsive for wavelengths slightly shorter than the resonance when the polarization of the incident light is parallel to the axis of the dimer. This behavior can be utilized to generate a stable separation distance between the nanoparticles. In the Rayleigh limit, the equilibrium distance is uniquely determined by the real part of the particle polarizability and the wavelength of the incident light. The results suggest that near-field optical forces can be used to manipulate and organize plasmonic nanoparticles with a tunable spatial resolution in the nanometer regime. PMID- 19550771 TI - Period-one oscillation for photonic microwave transmission using an optically injected semiconductor laser. AB - Optically injected semiconductor laser under periodone oscillation is investigated as a source for photonic microwave transmission over fiber. The period-one nonlinear dynamics of an optically injected laser is studied for the purpose of minimizing the microwave power penalty induced by chromatic dispersion. Over a large range of injection strengths and frequency detunings, we first obtain the mapping of the period-one oscillation characteristics, including the microwave frequency, the microwave power, and the single sideband (SSB) characteristics of the optical spectrum. By accounting for the fiber chromatic dispersion, we calculate its effect on the optical spectrum and the associated microwave power penalty. A mapping of the minimum microwave power deliverable after the maximum penalty is obtained. The system is shown to be least susceptible to the penalty when operated under strong injection with the frequency detuned above the Hopf bifurcation line. Microwave frequency beyond six times the relaxation resonance frequency can be effectively transmitted. PMID- 19550772 TI - Hybrid LPFG/MEFPI sensor for simultaneous measurement of high-temperature and strain. AB - A hybrid fiber-optic sensor consisting of a long-period fiber grating (LPFG) and a micro extrinsic Fabry-Perot (F-P) interferometric (MEFPI) sensor is proposed and demonstrated for simultaneous measurement of high-temperature and strain. The LPFG written by using high-frequency CO(3+) laser pulses is used for high temperature measurement while the MEFPI sensor fabricated by using 157nm F(2) laser pulses is used for strain measurement under high temperature. The distinguishing feature of such a hybrid fiber-optic sensor is that it can stand for high temperature of up to 650 masculineC and achieve precise measurement of strain under high temperature conditions simultaneously. PMID- 19550773 TI - Analysis of maximum reach in WDM PON architecture based on distributed Raman amplification and pump recycling technique. AB - We analyze the performance of bidirectional WDM PON architecture which utilizes distributed Raman amplification and pump recycling technique. The maximum reach at data rates of 622 Mb/s and 1.25 Gb/s in the proposed WDM PON architecture is calculated by taking into account the effects of power budget, chromatic dispersion of transmission fiber, and Raman amplification-induced noises with a given amount of Raman pump power. From the result, the maximum reach for 622 Mb/s and 1.25 Gb/s signal transmission is calculated to be 65 km and 60 km with a Raman pump power of 700 mW, respectively. We also find that the calculated results agree well with the experimental results which were reported previously. PMID- 19550774 TI - Nanosecond switching of fiber Bragg gratings. AB - A FBG was written in a two-hole fiber with internal alloy electrodes. Nanosecond high current pulses cause metal expansion, increase birefringence and tune the gratings with a response time of 29 ns. This short length, low loss, all-spliced high-speed wavelength switching devices described here has potential use in Q switching fiber laser. PMID- 19550775 TI - Aperiodic computer-generated volume holograms improve the performance of amplitude volume gratings. AB - We study the performance of amplitude computer-generated volume holograms (CGVH) in terms of efficiency and angular/frequency selectivity. We compare CGVHs to interferometrically-recorded amplitude volume holograms. Theoretical results show that amplitude CGVHs can increase the efficiency as well as the angular and wavelength selectivity relative to optically recorded amplitude volume holograms. We fabricate the CGVHs using femtosecond laser pulse micromachining in the bulk of glass and demonstrate results consistent with the theory. These results show that aperiodic three-dimensional structures provide the degrees of freedom necessary to improve the performance of volume diffractive optics. They suggest that, under certain circumstances, a departure from the Bragg paradigm provides enhanced volume diffraction properties. PMID- 19550776 TI - Localization of light in a lamellar structure with left-handed medium : the Light Wheel. AB - A rigorous analysis of the contra-directional coupling between a left-handed waveguide and a right-handed waveguide shows that such a structure supports twin modes with complex (conjugate) propagation constants. Using evanescent coupling or a source inside one of the waveguides, the twin modes can be excited. In this case, light rotates inside the lamellar structure forming an exotic mode : the light wheel. The structure can finally be seen as a new type of cavity and can be used for beam reshaping. PMID- 19550777 TI - High speed in situ depth profiling of ultrafast micromachining. AB - We demonstrate real-time depth profiling of ultrafast micromachining of stainless steel at scan rates of 46 kHz. The broad bandwidth and high power of the light source allows for simultaneous machining and coaxial Fourier-domain interferometric imaging of the ablation surface with depth resolutions of 6 mum. Since the same light is used to machine as to probe, spatial and temporal synchronization are automatic. PMID- 19550778 TI - Directivity enhancement and deflection of the beam emitted from a photonic crystal waveguide via defect coupling. AB - We experimentally and numerically investigate the spatial distribution of the emission from a photonic crystal waveguide, coupled with defects, that are located at the output edge. Two defects that are located symmetrically enhance the directivity of the beam compared to that of a plain waveguide, as was reported in recently conducted theoretical work. We further demonstrate that a single defect deflects of the beam. By choosing the defect resonance that is close to the edge of the pass band of the waveguide, where the group velocity of the beam within the waveguide is slow, a significant amount of deflection can be achieved. PMID- 19550779 TI - Pump-induced, dual-frequency switching in a short-cavity, ytterbium-doped fiber laser. AB - Using a short linear cavity composed of a section of highly ytterbium-doped fiber surrounded by two fiber Bragg gratings, dual-frequency switching is achieved by tuning the pump power of the laser. The dual-frequency switching is generated by the thermal effects of the absorbed pump in the ytterbium-doped fiber. At each frequency, the laser shows single-longitudinal-mode behavior. In each single-mode regime, the optical signal-to-noise ratio of the laser is greater than 50 dB. The dual-frequency, switchable, fiber laser can be designed for various applications by the careful selection of the two gratings. PMID- 19550780 TI - Vector field mapping of local polarization using gold nanoparticle functionalized tips: independence of the tip shape. AB - We have measured local electric field vectors of local polarizaton on the nanoscale using gold nanoparticle functionalized tips as local field scatterers. In our experiments, the local field induces a dipole-moment in the gold nanoparticle functionalized tip, which then radiates into the far-field, transferring the full information about the local electric field from the near into the far field. The polarization characteristics of the scattered fields are analyzed using a conventional ellipsometry method. The tip dependent scattering function- the polarizability tensor- is fully determined by far field scattering measurements. Once the polarizability tensor for each tip is correctly accounted for in the data analysis, our results show that the finally determined local field polarization vectors are essentially independent of the tip shape. PMID- 19550781 TI - Modified discrete particle model of optical scattering in skin tissue accounting for multiparticle scattering. AB - We rigorously account for the effects of multiparticle light scattering from a fractal sphere aggregate in order to simulate the optical properties of a soft biological tissue, human skin. Using a computational method that extends Mie theory to the multisphere case, we show that multiparticle scattering significantly affects the computed optical properties, resulting in a reduction in both scattering coefficient and anisotropy for the wavelengths simulated, as well as a significantly enhanced forward peak in the simulated phase function. The model is extended to incorporate the contribution of Rayleigh scatterers, which we show is required to obtain reasonable agreement with experimentally measured optical properties of skin tissue. PMID- 19550782 TI - Micro-structured fiber Bragg gratings: optimization of the fabrication process. AB - This work has been devoted to present and demonstrate a novel approach for the fabrication of micro-structured fiber Bragg gratings (MSFBGs) with enhanced control of the geometric features and thus of the spectral properties of the final device. The investigated structure relies on the localized stripping of the cladding layer in a well defined region in the middle of the grating structure leading to the formation of a defect state in the spectral response. In order to fully explore the versatility of MSFBGs for sensing and communications applications, a technological assessment of the fabrication process aimed to provide high control of the geometrical features is required. To this aim, here, we demonstrate that the optimization of this device is possible by adopting a fabrication process based on polymeric coatings patterned by high resolution UV laser micromachining tools. The function of the polymeric coating is to act as mask for the HF based chemical etching process responsible for the cladding stripping. Whereas, UV laser micromachining provides a valuable method to accurately pattern the polymeric coating and thus obtain a selective stripping along the grating structure. Here, we experimentally demonstrate the potentiality of the proposed approach to realize reliable and cost efficient MSFBGs enabling the prototyping of advanced photonics devices based on this technology. PMID- 19550783 TI - Birefringence compensated AWG demultiplexer with angled star couplers. AB - A new approach to birefringence compensation in arrayed waveguide gratings (AWG) is proposed. The star couplers are designed according to Rowland circle construction with an oblique incident/diffraction angle, similar to the case of an echelle grating. Such an AWG design is more general and flexible, and the conventional AWG becomes its special case when the grating angle is zero. By appropriately designing the star coupler shape, the birefringence of the arrayed waveguides can be compensated by that of the slab waveguides. The details of the design method and simulation results are presented. PMID- 19550784 TI - Enhancing intensity of emitted light from a ring by incorporating a circular groove. AB - We fabricated a ring containing a single circular groove (RCG) on silver film and which was supported on a glass substrate. We found that by changing the mean radius of the circular groove, the light intensity emitted from the RCG can be modulated by using the scattering light from the circular groove. In addition, we also fabricated circular grooves with the same depth but of different widths so that we could examine the scattering light behavior of the grooves. Herein, we propose a theoretical model which takes into account the amplitude modulation of the cylindrical waves. Our results showed that our proposed model agreed well with the experimental results. PMID- 19550785 TI - Generation of broadband mid-infrared pulses from an optical parametric amplifier. AB - We report on the direct generation of broadband mid-IR pulses from an optical parametric amplifier. Several crystals with extended IR transparency, when pumped at 800 nm, display a broad phase-matching bandwidth around 1 mum, allowing for the generation of idler pulses spanning the 3-5 mum wavelength range. Using LiIO(3), we produce 2muJ pulses tunable in the 3-4 mum range with bandwidth supporting 30-fs transform-limited duration. PMID- 19550786 TI - Experimental and theoretical thermal analysis of a Hybrid Silicon Evanescent Laser. AB - In this work we present both experimental and theoretical thermal analysis of an electrically pumped hybrid silicon evanescent laser. Measurements of an 850 mum long Fabry-Perot structure show an overall characteristic temperature of 51 oC, an above threshold characteristic temperature of 100 oC, and a thermal impedance of 41.8 oC/W. Finite element analysis of the laser structure predicts a thermal impedance of 43.5 oC/W, which is within 5% of the experimental results. Using the overall characteristic temperature, above threshold characteristic temperature, and the measured thermal impedance, the continuous wave output power vs. current from the laser is simulated and is in good agreement with experiment. PMID- 19550787 TI - Broadband wavelength conversion at 40 Gb/s using long serpentine As(2)S(3) planar waveguides. AB - We demonstrate broadband wavelength conversion of a 40 Gb/s return-to-zero signal by cross-phase modulation in a newly developed chalcogenide glass waveguide based photonic chip. These new serpentine As(2)S(3) waveguides offer a nonlinear coefficient approximately 1700 W(-1)km(-1) with 5x lower propagation loss over a length of 22.5 cm which ensures the full propagation length contributes towards the nonlinear process. This reduces the peak operating power thereby allowing a x4 increase in the data rate compared with previous results. Spectral measurements show the device operates over a span of 40 nm while system measurements show just over 1 dB of power penalty at a bit-error rate of 10(-9). This is primarily due to the compact planar waveguide design which minimizes the effect of groupvelocity dispersion. PMID- 19550788 TI - Direct rotation-induced intensity modulation in circular Bragg micro-lasers. AB - The effect of rotation on the lasing properties of circular Bragg micro-lasers is studied. In addition to the wavelength detuning caused by the Sagnac shift, a direct influence of the rotation on the lasing threshold of the clockwise and counterclockwise rotating modes is observed. The employment of the new effect for the realization of micro-rotation-sensors is discussed. PMID- 19550789 TI - Optimization of FM spectroscopy parameters for a frequency locking loop in small scale CPT based atomic clocks. AB - We describe the optimization of a Frequency Locked Loop (FLL) in an atomic clock which is based on Coherent Population Trapping (CPT) in (87)Rb vapor using the D(2) transition. The FLL uses frequency modulation (FM) spectroscopy and we study the effect of FM parameters (modulation frequency and index) on the sensitivity and the signal to noise ratio of the feedback signal in the FLL. The clock which employs a small spherical glass cell containing (87)Rb atoms and a buffer gas, exhibits a short term stability of 3x10(-11)/ radicaltau. The long term relative frequency stability of the 10 MHz output is better than 10(-10) with a drift of 10(-11) per day. PMID- 19550790 TI - Generalized inverse lithography methods for phase-shifting mask design. AB - Optical proximity correction (OPC) and phase shifting masks (PSM) are resolution enhancement techniques (RET) used extensively in the semiconductor industry to improve the resolution and pattern fidelity of optical lithography. In this paper, we develop generalized gradient-based RET optimization methods to solve for the inverse lithography problem, where the search space is not constrained to a finite phase tessellation but where arbitrary search trajectories in the complex space are allowed. Subsequent mask quantization leads to efficient design of PSMs having an arbitrary number of discrete phases. In order to influence the solution patterns to have more desirable manufacturability properties, a wavelet regularization framework is introduced offering more localized flexibility than total-variation regularization methods traditionally employed in inverse problems. The proposed algorithms provide highly effective four-phase PSMs capable of generating mask patterns with arbitrary Manhattan geometries. Furthermore, a double-exposure optimization method for general inverse lithography is developed where each exposure uses an optimized two-phase mask. PMID- 19550791 TI - Simultaneous demonstration on all-optical digital encoder and comparator at 40 Gb/s with semiconductor optical amplifiers. AB - We proposed and experimental demonstrated all-optical two line-four line encoder and two bit-wise comparator of RZ data streams at 40Gb/s based on cross gain modulation (XGM) and four wave mixing (FWM) in three parallel SOAs. Five logic functions for digital encoder and comparator between two signals A and B: AB, AB, AB, AB and AOmicronB, were achieved simultaneously. The first three optical logics are realized based on XGM in SOAs, the fourth is realized with FWM, and the fifth is the mixing result of the first and the fourth. A detuning filter is employed to improve the output performance. The output extinction ratio (ER) for the XGM operation is above 10dB, and the ER for FWM operation is around 8 dB. Wide and clear eye patterns for the five logic outputs can be observed. PMID- 19550792 TI - Extrusion of complex preforms for microstructured optical fibers. AB - We report a significant advance in preform extrusion and die design, which has allowed for the first time the fabrication of complex structured preforms using soft glass and polymer billets. Structural preform distortions are minimized by adjustment of the material flow within the die. The low propagation loss of an extruded complex bismuth glass fiber demonstrates the potential of this advanced extrusion technique for the fabrication of novel soft glass and polymer microstructured fiber designs. PMID- 19550793 TI - Chromatic discrimination by use of computer controlled set of light-emitting diodes. AB - We propose a novel approach for measurements of two-dimensional distribution of the reflection spectra with high spatial resolution. It is based on a subspace vector model of surface reflections and includes sequential illumination of the object by basis functions preliminary calculated with principal component analysis. A simple optical system consisting of a computer controlled set of light-emitting diodes and a photo-receiver operating in integration regime is used to acquire spatial distribution of reflection spectra in compressed form. The compressed data can be directly used for accurate color classification or recognition. The system's ability to distinguish metameric samples with extremely small hue difference is experimentally demonstrated. PMID- 19550794 TI - Sidestream Dark Field (SDF) imaging: a novel stroboscopic LED ring-based imaging modality for clinical assessment of the microcirculation. AB - Sidestream Dark Field (SDF) imaging, a stroboscopic LED ring-based imaging modality, is introduced for clinical observation of the microcirculation. SDF imaging is validated by comparison to Orthogonal Polarization Spectral imaging. Nailfold capillary diameters and red blood cell velocities were measured using both techniques and equal quantitative results were obtained. An image quality system was developed to quantitatively compare the quality of sublingually acquired microcirculatory images using OPS and SDF imaging. Venular contrast, sharpness, and quality were shown to be comparable for OPS and SDF imaging. However, capillary contrast and quality were shown to be significantly higher using SDF imaging. Venular granularity, in addition, was more clearly observable using SDF imaging. PMID- 19550795 TI - High speed engine gas thermometry by Fourier-domain mode-locked laser absorption spectroscopy. AB - We present a novel method for low noise, high-speed, real-time spectroscopy to monitor molecular absorption spectra. The system is based on a rapidly swept, narrowband CW Fourier-domain mode-locked (FDML) laser source for spectral encoding in time and an optically time-multiplexed split-pulse data acquisition system for improved noise performance and sensitivity. An acquisition speed of ~100 kHz, a spectral resolution better than 0.1 nm over a wavelength range of ~1335-1373 nm and a relative noise level of ~5 mOD (~1% minimum detectable base-e absorbance) are achieved. The system is applied for crank-angle-resolved gas thermometry by H(2)O absorption spectroscopy in an engine motoring at 600 and 900 rpm with a precision of ~1%. Influences of various noise sources such as laser phase and intensity noise, trigger and synchronization jitter in the electronic detection system, and the accuracy of available H(2)O absorption databases are discussed. PMID- 19550796 TI - Normalization detection scheme for high-speed optical frequency-domain imaging and reflectometry. AB - We introduce a new signal detection method that can effectively suppress the effect of relative intensity noise (RIN) in optical frequency-domain reflectometry or imaging (OFDR/OFDI) schemes to enhance the sensitivity and dynamic range. In this method, spectral interferogram signal is normalized digitally by a spectral reference signal that contains the real-time spectrum and the RIN information of the frequency-swept source. Unlike the conventional balanced detection method that suppresses only additive intensity noises, we found that our proposed scheme removes both the additive and convolutional contributions of the RINs in the final interferogram signals. Experimental demonstrations were performed using a stretched-pulse optical coherence tomography (SP-OCT) system where the high RIN of a supercontinuum source had been a serious drawback. We have experimentally verified the superiority of our proposed scheme in terms of its improved dynamic range in comparison to the balanced detection method. In addition, we have shown that the noise suppression performance is immune to the spectral imbalance characteristics of the optical components used in the system, whereas the common-mode noise rejection of the conventional balanced detection method is influenced by them. PMID- 19550797 TI - Double component long period waveguide grating filter in sol-gel material. AB - An efficient, tunable Long Period Waveguide Grating (LPWG) filter based on a new hybrid sol-gel material is demonstrated. The LPWG exhibits an attenuation of -22 dB and a high temperature sensitivity of ~3.3 nm/ degrees C. At room temperature the device shows an almost polarization independent wavelength. We took the advantage of the UV-curable sol-gel materials and used soft lithography to demonstrate a simple approach of integrating two LPWG filters on the same structure. The gratings were fabricated on the top and on the bottom of the same ridge waveguide and operate at communication wavelengths of 1550 and 1310 nm, respectively. PMID- 19550798 TI - Analyses of cladding modes in photonic crystal fiber. AB - Characteristics of cladding modes in a photonic crystal fiber (PCF) are numerically analyzed using the plane wave expansion method. The presence of the outer silica ring in the PCF tends to push the optical fields of the cladding modes toward the rim of the PCF, which creates 'ring modes' whose fields are tightly confined in the outer ring. The dispersion of the cladding modes are determined mainly by the dispersive property of the holey cladding structure. The optical field patterns of the cladding modes and the beatlengths between the fundamental mode and the cladding modes are also investigated. PMID- 19550799 TI - The effects of thermal equilibrium and contrast in LWIR polarimetric images. AB - Long-wave infrared (LWIR) polarimetric signatures provide the potential for day night detection and identification of objects in remotely sensed imagery. The source of optical energy in the LWIR is usually due to thermal emission from the object in question, which makes the signature dependent primarily on the target and not on the external environment. In this paper we explore the impact of thermal equilibrium and the temperature of (unseen) background objects on LWIR polarimetric signatures. We demonstrate that an object can completely lose its polarization signature when it is in thermal equilibrium with its optical background, even if it has thermal contrast with the objects that appear behind it in the image. PMID- 19550800 TI - Spectral shearing interferometry with spatially chirped replicas for measuring ultrashort pulses. AB - A new spectral shearing interferometry technique for ultrashort pulse characterization is demonstrated. The method makes use of a spectral shear that varies across the test pulse beam profile to generate a two-dimensional interferogram that allows simultaneous reference phase measurement and pulse field reconstruction from a single data set. The method uses a new configuration for upconversion of a single (non-spatially chirped) test pulse with two spatially chirped ancillary pulses in a medium with a highly asymmetric phase matching function. This technique is particularly suited for spectral regions where second harmonic is much easier detectable that the fundamental wavelength, such as telecom band around 1.5 μm, since all the data are available from the measurement at the upconverted wavelength. The high degree of redundancy in the two-dimensional interferogram provides a built-in check of the consistency of the reconstruction. PMID- 19550801 TI - Anisotropy without tensors: a novel approach using geometric algebra. AB - The most widespread approach to anisotropic media is dyadic analysis. However, to get a geometrical picture of a dielectric tensor, one has to resort to a coordinate system for a matrix form in order to obtain, for example, the index ellipsoid, thereby obnubilating the deeper coordinate-free meaning of anisotropy itself. To overcome these shortcomings we present a novel approach to anisotropy: using geometric algebra we introduce a direct geometrical interpretation without the intervention of any coordinate system. By applying this new approach to biaxial crystals we show the effectiveness and insight that geometric algebra can bring to the optics of anisotropic media. PMID- 19550802 TI - 1240nm high-power GaInNAs laser diodes. AB - We have fabricated 1240nm GaInNAs high-power semi-conductor laser diodes. In pulsed operation 1000 mum x 100 mum laser diodes show record low threshold current densities of 174 Acm(-2). Continuous wave output powers exceeding 4.6 Watts at room temperature and 6.2 Watts at a heatsink temperature of -5 degrees C are obtained from 1300 mum x 200 mum devices. The maximum wallplug efficiency of the device exceeds 40 % and the internal quantum efficiency reaches 0.89. Preliminary lifetime tests were performed for about 1000 h and show stable high power operation. PMID- 19550803 TI - Automatic retinal blood flow calculation using spectral domain optical coherence tomography. AB - Optical Doppler tomography (ODT) is a branch of optical coherence tomography (OCT) that can measure the speed of a blood flow by measuring the Doppler shift impinged on the probing sample light by the moving blood cells. However, the measured speed of blood flow is a function of the Doppler angle, which needs to be determined in order to calculate the absolute velocity of the blood flow inside a vessel. We developed a technique that can extract the Doppler angle from the 3D data measured with spectral-domain OCT, which needs to extract the lateral and depth coordinates of a vessel in each measured ODT and OCT image. The lateral coordinates and the diameter of a blood vessel were first extracted in each OCT structural image by using the technique of blood vessel shadowgram, a technique first developed by us for enhancing the retinal blood vessel contrast in the en face view of the 3D OCT. The depth coordinate of a vessel was then determined by using a circular averaging filter moving in the depth direction along the axis passing through the vessel center in the ODT image. The Doppler angle was then calculated from the extracted coordinates of the blood vessel. The technique was applied in blood flow measurements in retinal blood vessels, which has potential impact on the study and diagnosis of blinding diseases like glaucoma. PMID- 19550804 TI - Shot noise limited heterodyne detection of CARS signals. AB - We demonstrate heterodyne detection of CARS signals using a cascaded phase preserving chain to generate the CARS input wavelengths and a coherent local oscillator. The heterodyne amplification by the local oscillator reveals a window for shot noise limited detection before the signal-to-noise is limited by amplitude fluctuations. We demonstrate an improvement in sensitivity by more than 3 orders of magnitude for detection using a photodiode. This will enable CARS microscopy to reveal concentrations below the current mMolar range. PMID- 19550805 TI - Angular momentum of multimode and polarization patterns. AB - We study the mechanical properties of a broad class of multimode and polarization light patterns, resulting from the interference and superposition of waves in helical modes. General local and global properties of energy and angular momentum (AM) are identified, in order to define the conditions to optimize the AM with increasing beam complexity. We show the possibility to engineer independently the local densities of optical AM and energy, opening the possibility of an experimental demonstration of their respective effects in light-matter interaction. Multimode Laguerre-Gaussian beams also allows us to tailor the local spin AM through the Gouy phase. PMID- 19550806 TI - Achievement of arbitrary bandwidth of a narrow bandpass filter. AB - By adjusting the coating parameters to vary the refractive index of the thin film material, we are able to fine tune the bandwidth of a narrow bandpass filter to an arbitrary value. The relation between the varied index Deltan and the maximum arbitrary bandwidth was analyzed. A 4-skip-0 bandpass filter for a 100 GHz DWDM system was designed and fabricated. In addition, the relation between the tolerance of the index and the bandwidth was also analyzed to avoid broadening or narrowing the bandwidth. The final results showed that the arbitrary bandwidth met the requirements very well. PMID- 19550807 TI - Localized field enhancements in fractal shaped periodic metal nanostructures. AB - Fractal shaped structures formed with a 100-nm-period square lattice of gold nanoparticles placed on a gold film are characterized by using far-field nonlinear scanning optical microscopy, in which two-photon photoluminescence (TPL) excited with a strongly focused laser beam (in the wavelength range of 730 790 nm) is detected. The TPL images recorded for all wavelengths exhibit diffraction-limited (~ 0.6 mum) bright spots corresponding to the field intensity enhancement of up to 150, whose positions are dictated by the incident light wavelength and polarization. We relate these field enhancements to the occurrence of constructive interference of surface plasmons (SPs), which are excited by the incident radiation (due to scattering by nanoparticles) and partially reflected by fractal shaped boundaries due to a difference in the SP effective index at a flat and periodically corrugated gold surface. The conjecture on SP index difference is verified with observations (using leakage radiation microscopy) of SP focusing by circular and waveguiding by rectangular areas filled with periodically arranged nanoparticles. PMID- 19550808 TI - Supercontinuum generation in silicon photonic wires. AB - We observe spectral broadening of more than 350 nm, i.e., a 3/10-octave span, upon propagation of ultrashort 1.3-mum-wavelength optical pulses in a 4.7-mm-long silicon-photonic-wire waveguide. We measure the wavelength dependence of the spectral features and relate it to waveguide dispersion and input power. The spectral characteristics of the output pulses are shown to be consistent, in part, with higher-order soliton radiative effects. PMID- 19550809 TI - Adaptive Speckle Imaging Interferometry: a new technique for the analysis of microstructure dynamics, drying processes and coating formation. AB - We describe an extension of multi-speckle diffusing wave spectroscopy adapted to follow the non-stationary microscopic dynamics in drying films and coatings in a very reactive way and with a high dynamic range. We call this technique "Adaptive Speckle Imaging Interferometry". We introduce an efficient tool, the inter-image distance, to evaluate the speckle dynamics, and the concept of "speckle rate" (SR, in Hz) to quantify this dynamics. The adaptive algorithm plots a simple kinetics, the time evolution of the SR, providing a non-invasive characterization of drying phenomena. A new commercial instrument, called HORUS(R), based on ASII and specialized in the analysis of film formation and drying processes is presented. PMID- 19550810 TI - A simple method for determination of nonlinear propagation regimes in gases. AB - A simple method to evaluate the nonlinear propagation regimes in gases is demonstrated. The principle is to focus ultrashort laser pulses into a gas cell, vary the input pulse power and measure the transmission through a pinhole placed at the output. The resulting transmission curve yields an intuitive signature of various nonlinear propagation regimes. Going from low powers to higher, one first observes a brief decrease in the transmission due to nonlinear moving focus. Then, a sharp rise occurs, indicating the starting of the filamentation process. When the power increases further, the transmission saturates and eventually decreases due to the beginning of multi-filamentation. As a result, this method gives a quick and sensitive measurement of pulse energies required to have single and multiple filaments. PMID- 19550811 TI - Grating inscription in picosecond regime in thin films of functionalized DNA. AB - Polymers containing azo-benzene groups are useful holographic recording materials. In these materials the efficient mixed amplitude and phase gratings, frequently accompanied with photoinduced-surface relief gratings, can be inscribed with polarized laser light. The light-induced trans-cis-trans photoisomerization of azo-benzene groups is responsible for optical anisotropy in such systems. The aim of the present work is to study the dynamics of grating inscription in Disperse Red 1 doped deoxyrbonucleic acid- hexadecyltrimethylammonium material (DR1-DNACTMA) using 16 ps laser pulses (532 nm, 1.3 mJ). Results are compared with that obtained for other polymeric matrices loaded with DR1. The dynamics of the grating growth, due to repeated pulses from picosecond laser with 10 Hz repetition rate, was probed by measuring the intensity of the first order of diffraction of a cw He-Ne. We report results in function of the light polarization of writing beams. In this paper we present the first results of the grating inscription in functionalized DNA (in the picosecond pulse regime). PMID- 19550812 TI - Coherent and incoherent radial THz radiation emission from femtosecond filaments in air. AB - We show that the THz radiation emitted in the radial direction by a femtosecond filament created in air is linearly polarized and coherent. By applying an electric field along the filament axis this THz radiation is strongly enhanced and becomes incoherent and not polarized. PMID- 19550813 TI - Design of a maximally flat optical low pass filter using plasmonic nanostrip waveguides. AB - In this paper, we present a new optical range low pass filter based on plasmonic nanostrip waveguides. We calculate the characteristic impedance of plasmonic nanostrip waveguides and compare it with that of microstrip transmission lines. An optical range maximally flat low pass filter with subwavelength dimensions is designed based on the nanostrip waveguide structure. Finite-difference time domain (FDTD) simulations of the designed optical range filter are presented, which demonstrate subwavelength light confinement as well as acceptable filter cutoff performance. PMID- 19550814 TI - Measurement and compensation of optical aberrations using a single spatial light modulator. AB - We describe a compact adaptive optical system using a spatial light modulator (SLM) as a single element to both measure and compensate optical aberrations. We used a low-cost, off-the-shelf twisted nematic liquid-crystal display (TNLCD) optimally configured to achieve maximum phase modulation with near constant transmittance. The TNLCD acts both as the microlens array of a Hartmann-Shack wavefront sensor and as the aberration compensation element. This adaptive setup is easy to implement and offers great versatility. PMID- 19550815 TI - Pupil stabilization for SPHERE's extreme AO and high performance coronagraph system. AB - We propose a new concept of pupil motion sensor for astronomical adaptive optics systems and present experimental results obtained during the first laboratory validation of this concept. Pupil motion is an important issue in the case of extreme adaptive optics, high contrast systems, such as the proposed Planet Finder instruments for the ESO and Gemini 8-meter telescopes. Such high contrast imaging instruments will definitively require pupil stabilization to minimize the effect of quasi-static aberrations. The concept for pupil stabilization we propose uses the flux information from the AO system wave-front sensor to drive in closed loop a pupil tip-tilt mirror located in a focal plane. A laboratory experiment validates this concept and demonstrates its interest for high contrast imaging instrument. PMID- 19550816 TI - Ultrafast beam shaping with high-numerical-aperture microscope objectives. AB - Ultrafast diffraction results in spatiotemporal un-coupling of the wave field, inducing spectral anomalies and pulse stretching. Localized compensation may be achieved via angular dispersion driven by diffractive optical elements (DOEs). We report on an DOEs-based beam shaper of ultrashort optical pulses with high spatiotemporal resolution. Inspection of the validity of our approach is performed in the single-cycle regime. PMID- 19550817 TI - Two-dimensional photonic aperiodic crystals based on Thue-Morse sequence. AB - We investigate from a theoretical point of view the photonic properties of a two dimensional photonic aperiodic crystal. These structures are obtained by removing the lattice points from a square arrangement, following the inflation rules emerging from the Thue-Morse sequence. The photonic bandgap analysis is performed by means of the density of states calculation. The mechanism of bandgap formation is investigated adopting the single scattering model, and the Mie scattering. The electromagnetic field distribution can be represented as quasi-localized states. Finally, a generalized method to obtain aperiodic photonic structures has been proposed. PMID- 19550818 TI - High-efficiency, broad band, high-damage threshold high-index gratings for femtosecond pulse compression. AB - High efficiency, broad-band TE-polarization diffraction over a wavelength range centered at 800 nm is obtained by high index gratings placed on a non-corrugated mirror. More than 96% efficiency wide band top-hat diffraction efficiency spectra, as well as more than 1 J/cm(2) damage threshold under 50 fs pulses are demonstrated experimentally. This opens the way to high-efficiency Chirped Pulse Amplification for high average power laser machining by means of all-dielectric structures as well as for ultra-short high energy pulses by means of metal dielectric structures. PMID- 19550819 TI - Parasitic lasing suppression in high gain femtosecond petawatt Ti:sapphire amplifier. AB - New parasitic lasing suppression techniques are developed and high gain amplification is demonstrated in a petawatt level Ti:sapphire amplifier based on the chirped pulse amplification (CPA) scheme. Cladding the large aperture Ti:sapphire with refractive-index matched liquid doped with absorber suppresses the transverse lasing. The acousto-optic programmable dispersive filter (AOPDF) is used to realize side-lobe suppression in the temporal profile of the compressed pulse. The 800 nm laser output with peak power of 0.89 PW and pulse width of 29.0 fs is demonstrated. PMID- 19550820 TI - Slow to superluminal light waves in thin 3D photonic crystals. AB - Phase measurements on self-assembled three-dimensional photonic crystals show that the group velocity of light can flip from small positive (slow) to negative (superluminal) values in samples of a few mum size. This phenomenon takes place in a narrow spectral range around the second-order stop band and follows from coupling to weakly dispersive photonic bands associated with multiple Bragg diffraction. The observations are well accounted for by theoretical calculations of the phase delay and of photonic states in the finite-sized systems. PMID- 19550821 TI - Enhanced phase-matching for generation of soft X-ray harmonics and attosecond pulses in atomic gases. AB -

Focus Serial: Frontiers of Nonlinear Optics

We theoretically investigate the generation of high harmonics and attosecond pulses by mid-infrared (IR) driving fields. Conditions for coherent build-up of high harmonics are revisited. We show that the coherence length dictated by ionization-induced dephasing does not constitute an ultimate limitation to the coherent growth of soft X-ray (> 100 eV) harmonics driven by few-cycle mid-IR driving pulses: perfect phase-matching, similar to non-adiabatic self-phase- matching, can be achieved even without non-linear deformation of the driving pulse. Our trajectory-based analysis of phase-matching reveals several important advantages of using longer laser wavelengths: conversion efficiency can be improved by orders of magnitude, phase-matched build-up of harmonics can be achieved in a jet with a high gas pressure, and isolated attosecond pulses can be extracted from plateau harmonics. PMID- 19550822 TI - Nonlinear waveguide optics and photonic crystal fibers. AB -

Focus Serial: Frontiers of Nonlinear Optics

Optical fibers and waveguides provide unique and distinct environments for nonlinear optics, because of the combination of high intensities, long interaction lengths, and control of the propagation constants. They are also becoming of technological importance. The topic has a long history but continues to generate rapid development, most recently through the invention of the new forms of optical fiber collectively known as photonic crystal fibers. Some of the discoveries and ideas from the new fibers look set to have lasting influence in the broader field of guided-wave nonlinear optics. In this paper we introduce some of these ideas. PMID- 19550823 TI - A wavelength-tunable fiber-coupled source of narrowband entangled photons. AB - We demonstrate a wavelength-tunable, fiber-coupled source of polarization- entangled photons with extremely high spectral brightness and quality of entanglement. Using a 25 mm PPKTP crystal inside a polarization Sagnac interferometer we detect a spectral brightness of 273000 pairs (s mW nm)(-1), a factor of 28 better than comparable previous sources while state tomography showed the two-photon state to have a tangle of T = 0.987. This improvement was achieved by use of a long crystal, careful selection of focusing parameters and single-mode fiber coupling. We demonstrate that, due to the particular geometry of the setup, the signal and idler wavelengths can be tuned over a wide range without loss of entanglement. PMID- 19550825 TI - Implications of higher-order mode content in large mode area fibers with good beam quality. AB - The impact of Higher-Order Mode content on beam quality in large mode area fibers supporting several guided modes is carefully investigated. It is shown that even excellent beam quality (M(2) < 1.1) in LMA fibers does not guarantee low HOM content, and that the presence of HOMs can lead to significant uncontrollable changes in beam quality, peak intensity, and pointing uncertainty that depend on the uncontrollable relative phase of the modes in the fiber. PMID- 19550824 TI - The von Neumann picture: a new representation for ultrashort laser pulses. AB - In recent years, the use of joint time-frequency representations to characterize and interpret shaped femtosecond laser pulses has proven to be very useful. However, the number of points in a joint time-frequency representation is daunting as compared with those in either the frequency or time representation. In this article we introduce the use of the von Neumann representation, in which a femtosecond pulse is represented on a discrete lattice of evenly spaced time frequency points using a non-orthogonal Gaussian basis. We show that the information content in the von Neumann representation using a lattice of radicalN points in time and radicalN points in frequency is exactly the same as in a frequency (or time) array of N points. Explicit formulas are given for the forward and reverse transformation between an N-point frequency signal and the von Neumann representation. We provide numerical examples of the forward and reverse transformation between the two representations for a variety of different pulse shapes; in all cases the original pulse is reconstructed with excellent precision. The von Neumann representation has the interpretational advantages of the Husimi representation but requires a bare minimum number of points and is stably and conveniently inverted; moreover, it avoids the periodic boundary conditions of the Fourier representation. PMID- 19550826 TI - Optimal design of computer-generated holograms to minimize sensitivity to fabrication errors. AB - Aspheric optical surfaces are often tested using computer-generated holograms (CGHs). The etching of the CGH pattern must be highly accurate to create desired wavefronts. Variations of line width, etching depth, and surface roughness cause unwanted wavefront errors. The sensitivity to these manufacturing errors is studied using scalar diffraction analysis. We provide a parametric model that can be used for optimizing the CGH design to give good diffraction efficiency and limited sensitivity to manufacturing errors. PMID- 19550827 TI - Four-wave mixing based widely tunable wavelength conversion using 1-m dispersion shifted bismuth-oxide photonic crystal fiber. AB - We demonstrate widely tunable wavelength conversion based on four-wave mixing using a dispersion-shifted bismuth-oxide photonic crystal fiber (Bi-PCF). A 1 meter-long Bi-PCF is used as the nonlinear medium for wavelength conversion of a 10 Gb/s non-return-to-zero (NRZ) signal. A 3- dB working range of the converted signal over 35 nm is obtained with around 1-dB power penalty in the bit-error rate measurements. PMID- 19550828 TI - Computer-guided alignment II :Optical system alignment using differential wavefront sampling. AB - We present a differential wavefront sampling method for the efficient alignment of centred optical systems. Using the inter-element effects reported in our previous study, this method generates a linear symmetric matrix that relates the optical wavefront to misalignments within the system. The solution vector of this matrix equation provides a unique description of decentre and tilt misalignments of the system. We give a comparison of this approach to the existing method in the first case study and then illustrate characteristics of the new approach using the subsequent four case studies and Monte-Carlo alignment simulations. The results reveal superiority of the method over the existing one in misalignment estimation accuracy and demonstrate the practical feasibility and robustness. PMID- 19550829 TI - Self compression of Yb-doped solid-state lasers by combination of self-phase modulation and group-velocity dispersion in KDP crystal. AB - We propose and evaluate numerically the self compression for Yb-doped solid-state laser pulses in a KDP crystal by the combination of the group-velocity dispersion and the self-phase modulation. The self compression is achievable as the group velocity-dispersion coefficient of KDP crystals is negative around a 1-mum wavelength. Numerical results showed that the laser pulse in the range 50-200 fs can be compressed as short as 12.8 fs. This self-compression method is simple and low cost, which is possible to be applied to an Yb-doped solid-state laser system with a large-scale beam and an ultrahigh intensity in the regime of tens of femotoseconds. PMID- 19550830 TI - Flame front detection and characterization using conditioned particle image velocimetry (CPIV). AB - We investigate the ability of the conditioned particle image velocimetry technique (CPIV) to derive the actual flame front position in turbulent premixed flames. In CPIV, the flame front shape is deduced from the step in the particle number density in PIV images caused by the steep temperature increase in the reaction zone of premixed flames. In a validation experiment the true flame front position is deduced for comparison from simultaneous heat release measurements using planar LIF measurements of OH and CH(2)O. It is found that CPIV yields nearly the same spatial position as the heat release measurements or the steepest slope in the OH distribution. Furthermore, statistical quantities, derived from the extracted flame front shape, like the spatially resolved turbulent flux, the flame surface density and the flame front curvature are compared, showing negligible differences between the applied methods. PMID- 19550831 TI - Color filter based on a subwavelength patterned metal grating. AB - A color filter incorporating a subwavelength patterned grating in a metal film perforated with a square array of circular apertures on a quartz substrate was accomplished. Its performance was enhanced by applying a dielectric overlay to the grating layer so as to match the refractive indices of the media on either side of it. The device was designed by utilizing the finite-difference time domain method and implemented by adopting the electron-beam direct-writing technique. Two different devices were fabricated with the structural parameters: the grating height of 50 nm and the pitch of 340 nm for the red color and 260 nm for the green color. For the red color filter the center wavelength was 680 nm and the peak transmission 57%, while for the green color one the center wavelength was 550 nm and the peak transmission 50%. It was confirmed the introduction of the index matching overlay led to an increase of ~15% in the transmission efficiency and helped combine double bands into a single dominant band as well, thereby improving the color selectivity of the filter. PMID- 19550832 TI - Processing advantages of linear chirped fiber Bragg gratings in the time domain realization of optical frequency-domain reflectometry. AB - The inclusion of a linear chirped fiber Bragg grating for short pulse dispersion is shown to enhance the time domain realization of optical frequency-domain reflectometry. A low resolution demonstrator is constructed with single surface scans containing 140 resolvable spots. The system dynamic range meets that shown in earlier demonstrations without digital post-processing for signal linearization. Using a conjugate pair of chirped pulses created by the fiber grating, ranging is performed with position and velocity information decoupled. Additionally, by probing the target with short pulses and introducing grating dispersion just before photodetection, velocity immune ranging is demonstrated. PMID- 19550833 TI - Second-harmonic generation by an astigmatic partially coherent beam. AB - We investigate second-harmonic generation by an astigmatic partially coherent beam. An explicit expression for the second-order correlation function of the second-harmonic field is obtained. The properties of the generated field and the conversion efficiency for second-harmonic generation are studied numerically. We find that using an astigmatic instead of a stigmatic partially coherent pump beam can increase the conversion efficiency of the second-harmonic generation. PMID- 19550834 TI - Fluorescence ratio thermometry in a microfluidic dual-beam laser trap. AB - The dual-beam laser trap is a versatile tool with many possible applications. In order to characterize its thermal properties in a microfluidic trap geometry we have developed a non-intrusive fluorescence ratio technique using the temperature sensitive dye Rhodamine B and the temperature independent reference dye Rhodamine 110. We measured temperature distribution profiles in the trap with submicron spatial resolution on a confocal laser-scanning microscope. The maximum heating in the center of the trap amounts to (13 +/- 2) degrees C/W for a wavelength of lambda = 1064 nm and scales linearly with the applied power. The measurements correspond well with simulated temperature distributions. PMID- 19550835 TI - A photonic crystal ring resonator formed by SOI nano-rods. AB - The design, fabrication and measurement of a silicon-on-insulator (SOI) two dimensional photonic crystal ring resonator are demonstrated in this study. The structure of the photonic crystal is comprised of silicon nano-rods arranged in a hexagonal lattice on an SOI wafer. The photonic crystal ring resonator allows for the simultaneous separation of light at wavelengths of 1.31 and 1.55mum. The device is fabricated by e-beam lithography. The measurement results confirm that a 1.31mum/1.55mum wavelength ring resonator filter with a nano-rod photonic crystal structure can be realized. PMID- 19550836 TI - Hemodynamic responses to antivascular therapy and ionizing radiation assessed by diffuse optical spectroscopies. AB - Diffuse optical methods were used to monitor two different therapies in K1735 malignant mouse melanoma tumor models: anti-vascular therapy and radiation therapy. Anti-vascular therapy induced acute variation in hemodynamic parameters within an hour, and radiation therapy induced longitudinal changes within 2 weeks. During anti-vascular therapy, the drug Combretastatin A-4 3-O-Phosphate (CA4P, 2.5 mg/200 mul PBS/mouse) significantly decreased tissue blood flow (65%) and blood oxygenation (38%) one hour after injection. In the longitudinal study, single-fraction ionizing radiation (12 Gy x 1) induced significant reduction of tissue blood flow (36%) and blood oxygenation (24%) 14 days after radiation. The results correlated well with contrast enhanced ultrasound, tumor histology, and a nitroimidazole hypoxia marker (EF5). The research provides further evidence that noninvasive diffuse optical spectroscopies can be useful tools for monitoring cancer therapy in vivo. PMID- 19550838 TI - Versatile opto-fluidic ring resonator lasers with ultra-low threshold. AB - We develop a versatile integrated opto-fluidic ring resonator (OFRR) dye laser that can be operated regardless of the refractive index (RI) of the liquid. The OFRR is a micro-sized glass capillary with a wall thickness of a few micrometers. When the liquid in the core has an RI lower than that of the capillary wall (n=1.45), the capillary circular cross-section forms the ring resonator and supports the whispering gallery modes (WGMs) that interact evanescently with the gain medium in the core. When the core RI is higher than that of the wall, the WGMs exist at the core/wall interface. In both cases, the WGMs can have extremely high Q-factor (>109), providing excellent optical feedback for low-threshold lasing. In this paper, we analyze the OFRR laser for various core RI's and then we demonstrate the R6G laser when the dye is in ethanol (n=1.36), chloroform (n=1.445), and quinoline (n=1.626). The lasing threshold of 25 nJ/mm(2) is achieved, two to three orders of magnitude lower than the previous work in microfluidic lasers. We further show that the laser emission can be efficiently out-coupled via an optical waveguide in touch with the OFRR for both high and low RI liquid core, allowing for easy guiding and delivery of the laser light. PMID- 19550837 TI - Complete experimental characterization of stimulated Brillouin scattering in photonic crystal fiber. AB - We provide a complete experimental characterization of stimulated Brillouin scattering in a 160 m long solid-core photonic crystal fiber, including threshold and spectrum measurements as well as position-resolved mapping of the Brillouin frequency shift. In particular, a three-fold increase of the Brillouin threshold power is observed, in excellent agreement with the spectrally-broadened Brillouin gain spectrum. Distributed measurements additionally reveal that the rise of the Brillouin threshold results from the broadband nature of the gain spectrum all along the fiber and is strongly influenced by strain. Our experiments confirm that these unique fibers can be exploited for the passive control or the suppression of Brillouin scattering. PMID- 19550839 TI - Near-field studies of microwave three-dimensional photonic crystals with waveguides. AB - By utilizing a vector network analyzer, the field distributions on the surface of a three-dimensional woodpile photonic crystal with a straight waveguide or a bend waveguide buried under the surface were measured in the microwave regime. The information of field profile and propagation characteristics of the guided modes can be successfully extracted from the surface near-field measurement. This work indicates that the near-field detection can become a promising means for experimental characterization of three-dimensional photonic crystal devices in supplement to the usual transmission spectrum measurement. PMID- 19550841 TI - Reflectance reconstruction for multispectral imaging by adaptive Wiener estimation. AB - In multispectral imaging, Wiener estimation is widely adopted for the reconstruction of spectral reflectance. We propose an improved reflectance reconstruction method by adaptively selecting training samples for the autocorrelation matrix calculation in Wiener estimation, without a prior knowledge of the spectral information of the samples being imaged. The performance of the proposed adaptive Wiener estimation and the traditional method are compared in the cases of different channel numbers and noise levels. Experimental results show that the proposed method outperforms the traditional method in terms of both spectral and colorimetric prediction errors when the imaging channel number is 7 or less. When the imaging system consists of 11 or more channels, the color accuracy of the proposed method is slightly better than or becomes close to that of the traditional method. PMID- 19550840 TI - Ultrashort pulse Yb:LaSc(3)(BO(3))(4) mode-locked oscillator. AB - Passive mode-locked laser operation based on an Yb-doped lanthanum scandium borate crystal is demonstrated. Pulse durations as short as 58 fs and 67 fs were achieved applying a Ti:sapphire- and a diode-laser pump source, respectively. The average output powers were 73 mW and 39 mW at a repetition rate of 90 MHz. The laser was broadly tunable from 1028 to 1057 nm in the sub-200 fs pulse regime. PMID- 19550842 TI - Spectral characteristics and bend response of Bragg gratings inscribed in all solid bandgap fibers. AB - In this paper, we investigate the spectral characteristics and bend response of fiber Bragg gratings (FBGs) in all-solid photonic bandgap fibers (PBGFs). We inscribe FBGs within the secondary bandgap by ultraviolet (UV) side illumination and observe the couplings to backward core mode, guided LP(01) and LP(11) supermodes and radiative LP(02) supermodes. The mechanisms of these resonant couplings in the FBG are described in detail. We demonstrate that only those supermodes with certain phase relationships and symmetric mode field profiles are responsible for the supermode resonances. When the fiber grating is bent, the guided supermode resonances become chirped as a result of the strain gradient over the fiber cross section. Meanwhile, the core resonance is enhanced, due to more energy of the core mode distributed in the cladding rods. The bend response is direction dependant owing to the nonuniform UV-induced average index raises and index modulation over the high-index rod lattice. . PMID- 19550843 TI - Fast full-view photoacoustic imaging by combined scanning with a linear transducer array. AB - We present a fast full-view photoacoustic imaging system for visualizing tissue structures using a linear transducer array with combined scan. In this system, a 128-element linear transducer array was used to detect photoacoustic signals by combined scanning of electronic scan and mechanical scan. An improved limited field filtered back projection algorithm with directivity factors was applied to reconstruct the optical absorption distribution. The experiments of phantoms and in vivo blood vessels in a rat brain were performed with this system. And a clear view of the curve boundaries of objects and the network of blood vessels of rat's brain were acquired. The experimental results demonstrate the multi-element photoacoustic imaging system has the ability of imaging complicated structures of objects. PMID- 19550844 TI - A novel Zn-indiffused mode converter in x-cut lithium niobate. AB - A novel Zn-indiffused mode converter has been proposed and experimentally studied in an x-cut/z-propagation lithium niobate at a wavelength of 0.632 mum for the first time. The optimized phase-matching and mode-conversion voltages for maximum conversion are 12 V and -?5 V, respectively. The results show that the proposed mode converter can operate with a stable conversion efficiency of about 99.5% between TM and TE polarizations at a throughput power of 25 muW in a period of 60 min. Moreover, a comparison of optical power-handling stability between the Ti indiffused and the Zn-indiffused channel waveguides, was explored. The encouraging results indicate that the Zn-indiffused waveguide has better power stability than the Ti-indiffused waveguide. Thus, it is expected that the proposed mode converter will have better stability than the conventional Ti indiffused ones, especially in the visible wavelength region. PMID- 19550845 TI - Terahertz movie of internal transmission imaging. AB - Recently terahertz imaging using two-dimensional E-O sampling has attracted much interest because it can acquire real-time terahertz images unlike a conventional raster scan method. We are applying this technique to the non-destructive measurement of opaque materials in a visible range. We acquired 10-fps consecutive terahertz transmission images: dripping water in a plastic pipe and metal included in a piece of gum. Since the obtained images were confirmed to be proportional to the electric field of the terahertz waves, the images in the present paper are useful for quantitative analysis. We also showed the signal-to noise ratio of the terahertz images. PMID- 19550846 TI - Arbitrary-to-linear or linear-to-arbitrary polarization controller based on Faraday and Pockels effects in a single BGO crystal. AB - We propose an arbitrary-to-linear or linear-to-arbitrary polarization controller based on the mutual action of Faraday and Pockels effects in a single Bi(4)Ge(3)O(12) (BGO) crystal after the wave coupling theory describing these two effects. It is demonstrated that, the expected conversion of arbitrary-to-linear or linear-to-arbitrary polarization state of light can be realized by adjusting the applied electric and magnetic fields. PMID- 19550847 TI - Experimental and numerical study of pulse dynamics in positive net-cavity dispersion modelocked Yb-doped fiber lasers. AB - We report on environmentally stable mode-locked Yb-doped all-fiber lasers operating in the wave-breaking-free and stretched-pulse regime. The compact linear cavity is constructed with saturable absorber mirror directly glued to the fibers end-facet as nonlinear mode-locking mechanism and chirped fiber Bragg grating for dispersion management, thus, without any free-space optics. In the wave-breaking-free regime the laser generates positively-chirped pulses with a pulse duration of 15.4 ps. These pulses are compressed to 218 fs in a hollow-core photonic bandgap fiber spliced to the output port. Adaptation of dispersion management has led to operation in the stretched-pulse regime, where a parabolic spectral profile is obtained as well. In this regime pulses are compressible to 213 fs. Numerical simulations are presented which confirm the wave-breaking-free and stretched-pulse evolution inside the fiber laser cavity. Both regimes are compared in terms of pulse quality. PMID- 19550848 TI - Mapping the broadband polarization properties of linear 2D SOI photonic crystal waveguides. AB - Both quasi-TE and TM polarisation spectra for a silicon-on-insulator (SOI) waveguide are recorded over (1100-1700)nm using a broadband supercontinuum source. By studying both the input and output polarisation eigenstates we observe narrowband resonant cross coupling near the lowest quasi-TE mode cut-off. We also observe relatively broadband mixing between the two eigenstates to generate a complete photonic bandgap. By careful analysis of the output polarisation state we report on an inherent non-reciprocity between quasi TE and TM fundamental mode cross coupling. The nature of polarisation distinction in such bandgap structures is discussed in the context of polarisation scattering at an interface. PMID- 19550849 TI - Near infrared multifactor identification tags. AB - We propose a compact technique for encryption-verification that relies on the following elements: multifactor encryption, which permits the simultaneous verification of up to four factors; distortion-invariant ID tag for remote identification; near infrared (NIR) writing and readout of the ID tag signal for invisible transmission; and optical processor, based on joint transform pattern recognition by optical correlation, for automatic verification of information. A highly-reliable security system is obtained by joining the advantages of all these elements for the first time. A novel NIR ID tag is designed and built by using commonly available materials. The very ID tag content cannot be visually perceived at naked eye; it cannot be either copied, scanned, or captured by any conventional device. Experimental results based on the NIR ID tag are shown. The satisfactory results obtained demonstrate a new insight into the applications of the compact and efficient technique for high-secure identification systems. PMID- 19550850 TI - Diffraction by fractal metallic supergratings. AB - The reflectance of corrugated surfaces with a fractal distribution of grooves is investigated. Triadic and polyadic Cantor fractal distributions are considered, and the reflected intensity is compared with that of the corresponding periodic structure. The self-similarity property of the response is analyzed when varying the depth of the grooves and the lacunarity parameter. The results confirm that the response is self-similar for the whole range of depths considered, and this property is also maintained for all values of the lacunarity parameter. PMID- 19550851 TI - Photonic crystal fibers confining light by both index-guiding and bandgap guiding: hybrid PCFs. AB - We propose two kinds of hybrid photonic crystal fiber (PCF) structures and investigate the properties of such PCFs in detail. The modal effective index, mode field area, confinement loss, group velocity dispersion, and birefringence are numerically simulated and compared with those of the corresponding index guiding and bandgap PCFs, which allows for a deeper understanding of the guiding mechanism of the hybrid PCFs. The advantages of hybrid PCFs and potential applications are also discussed. PMID- 19550852 TI - Sensitized nonlinear emission of gold nanoparticles. AB - We have studied Stokes and anti-Stokes emission of Au nanoparticles suspended in pure methanol and methanol solution of rhodamine 6G dye. In the presence of dye, excitation of anti-Stokes emission of gold involves two-photon absorption in rhodamine 6G molecules followed by the energy transfer to Au nanoparticles with simultaneous absorption of one pumping photon by Au. The sensitization by dye molecules caused six-fold enhancement of the anti-Stokes emission of gold nanoparticles. PMID- 19550853 TI - High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events. AB - We present a time domain optically sectioned fluorescence lifetime imaging (FLIM) microscope developed for high-speed live cell imaging. This single photon excited system combines wide field parallel pixel detection with confocal sectioning utilizing spinning Nipkow disc microscopy. It can acquire fluorescence lifetime images of live cells at up to 10 frames per second (fps), permitting high-speed FLIM of cell dynamics and protein interactions with potential for high throughput cell imaging and screening applications. We demonstrate the application of this FLIM microscope to real-time monitoring of changes in lipid order in cell membranes following cholesterol depletion using cyclodextrin and to the activation of the small GTP-ase Ras in live cells using FRET. PMID- 19550854 TI - Mode scalability in bent optical fibers. AB - This paper introduces a simple, analytical method for generalizing the behavior of bent, weakly-guided fibers and waveguides. It begins with a comprehensive study of the modes of the bent step-index fiber, which is later extended to encompass a wide range of more complicated waveguide geometries. The analysis is based on the introduction of a scaling parameter, analogous to the V-number for straight step-index fibers, for the bend radius. When this parameter remains constant, waveguides of different bend radii, numerical apertures and wavelengths will all propagate identical mode field distributions, except scaled in size. This allows the behavior of individual waveguides to be broadly extended, and is especially useful for generalizing the results of numerical simulations. The technique is applied to the bent step-index fiber in this paper to arrive at simple analytical formulae for the propagation constant and mode area, which are valid well beyond the transition to whispering-gallery modes. Animations illustrating mode deformation with respect to bending and curves describing polarization decoupling are also presented, which encompass the entire family of weakly-guided, step-index fibers. PMID- 19550855 TI - Fluorescence component in the reflectance spectra from coastal waters. Dependence on water composition. AB - Based on HYDROLIGHT simulations of more than 2000 reflectance spectra from datasets typical of coastal waters with highly variable optically active constituents as well as on intercomparisons with field measurements, the magnitude of chlorophyll fluorescence was analyzed and parameterized as a function of phytoplankton, CDOM, and suspended inorganic matter concentrations. Using the parameterizations developed, we show that variations in the fluorescence component of water leaving radiance in coastal waters are due more to the variability of attenuation in the water than to the variability of the fluorescence quantum yield, which we estimate to be relatively stable at around 1%. Finally, the ranges of water conditions where fluorescence plays a significant role in the reflectance NIR peak and where it is effectively undetectable are also determined. PMID- 19550856 TI - The NIR-SWIR combined atmospheric correction approach for MODIS ocean color data processing. AB - A method of ocean color data processing using the combined near-infrared (NIR) and shortwave infrared (SWIR) bands for atmospheric correction for the Moderate Resolution Imaging Spectroradiometer (MODIS) on Aqua is proposed. MODIS-Aqua has been producing the high quality ocean color products in the open oceans, but there are still some significant errors in the derived products in the coastal regions. With the proposed NIR-SWIR combined algorithm, MODIS ocean color data can be processed using the standard (NIR) atmospheric correction algorithm for the open oceans, whereas for the turbid waters in the coastal region the SWIR atmospheric correction algorithm can be executed. The turbid water index developed by Shi and Wang (2007) (Remote Sens. Environ. 110, 149-161 (2007)) is computed prior to the atmospheric correction for the identification of the productive and/or turbid waters where the SWIR algorithm can be operated. For non turbid ocean waters (discriminated using the turbid water index criterion), the MODIS data are still processed using the standard (NIR) algorithm. The NIR-SWIR combined algorithm has been tested and evaluated. Two examples from MODIS-Aqua measurements along the U.S. and China east coast regions show improved ocean color products with the new approach. In particular, there are no obvious data discontinuities between using the NIR and SWIR methods. Therefore, with the NIR SWIR combined approach for the MODIS ocean color data processing, good quality ocean color products can be derived both in clear (open) oceans as well as for turbid coastal waters. PMID- 19550857 TI - Narrow-band filtering with whispering modes in gratings made of fibers. AB - We present a numerical study of whispering modes in gratings made of fibers. Due to the strong localization of the modes inside each fiber, it is possible to obtain narrow-band filters with very broad angular tolerance. PMID- 19550858 TI - Self-organized tungsten nanospikes grown on subwavelength ripples induced by femtosecond laser pulses. AB - We report on the formation of nanoscale tungsten spikes generated on subwavelength periodic ripples which built up by single beam 800 nm femtosecond laser pulses. The nanospikes have a diameter ranging from 10 to 100 nm and are up to 250 nm in length. The nanospikes orientate from the ridges toward the valleys of the ripple structures independent of the polarization of the light. The heterogeneous nucleation of the liquid phase at the irradiated surface and the inhomogeneous surface roughness are considered as the mechanism of this nanospike formation. PMID- 19550859 TI - Speckle noise reduction algorithm for optical coherence tomography based on interval type II fuzzy set. AB - A novel speckle reduction technique based on soft thresholding of wavelet coefficients using interval type II fuzzy system was developed for reducing speckle noise in Optical Coherence Tomography images. The proposed algorithm is an extension of a recently published method for filtering additive Gaussian noise by use of type I fuzzy system. Unlike type I, interval type II fuzzy based thresholding filter considers the uncertainty in the calculated threshold and the wavelet coefficient is adjusted based on this uncertainty. A single parameter controls the signal-to-noise (SNR) improvement. Application of this novel algorithm to optical coherence tomograms acquired in-vivo from a human finger tip show reduction in the speckle noise with little edge blurring and image SNR improvement of about 10dB. Comparison with adaptive Wiener and adaptive Lee filters, applied to the same image, demonstrated the superior performance of the fuzzy type II algorithm in terms of image metrics improvement. PMID- 19550860 TI - A compact system for simultaneous measurement of linear and angular displacements of nano-stages. AB - We report on a novel compact interferometery system for measuring parasitic motions of a precision stage. It is a combination of a Michelson interferometer with an auto-collimator, of which full physical dimension is mere 70 mm x80 mm x35 mm (WxLxH) including optical components, photo-detectors, and electronic circuits. Since the beams, which measure displacement and angle, can be directed at the same position on the moving mirror, the system is applicable for testing small nano-stages where commercial interferometers are not able to be used. And thus, errors from nano-scale deformation of the moving mirror can be minimized. We find that the residual errors of linear and angular motion measurements are 2.5 nm in peak-to-peak and 0.2'', respectively. PMID- 19550861 TI - High-efficiency and stable optical transmitter using VCSEL-direct-bonded connector for optical interconnection. AB - A high-efficiency optical transmitter module for PCB (printed circuit board) based interconnections was fabricated using a bottom-emitting VCSEL. The bottom emitting VCSEL was directly bonded by an epoxy on a 90 degrees -bent fiber connector which is inserted into the PCB to couple to the fiber layer embedded in the board. A ray trace simulation indicates that close contact between the VCSEL and the connector removes most of the losses due to Fresnel reflection and beam divergence. This tendency was experimentally identified. Thermal dissipation through the epoxy layer and the connector also improves significantly the power characteristics of the VCSEL. The VCSEL after bonding on the connector shows about 40% higher power compared to that of the bare VCSEL at the current showing a peak power before bonding. The results indicate that direct bonding improves both optical and electrical efficiencies. A successful eye diagram at the speed of 5 Gb/s/ch with 850 nm was accomplished from the VCSEL-direct-bonded transmitter module. PMID- 19550862 TI - Supercontinuum generation in an ultrafast laser inscribed chalcogenide glass waveguide. AB - The authors report supercontinuum generation in an ultrafast laser inscribed chalcogenide glass waveguide. The waveguides were fabricated using a Yb:glass cavity-dumped femtosecond oscillator with 600- kHz repetition rate. The waveguides were pumped using an optical parametric amplifier tuned to 1500 nm with a bandwidth of 100 nm. The broadest resulting supercontinuum spanned 600 nm (at -15 dB points) from 1320 to 1920 nm. The supercontinuum was generated in the normal dispersion regime, enhancing stability, and exhibits a smooth spectral shape. PMID- 19550863 TI - Analytical analysis of modulated signal in apertureless scanning near-field optical microscopy. AB - Eliminating background-scattering effects from the detected signal is crucial in improving the performance of super-high-resolution apertureless scanning near field optical microscopy (A-SNOM). Using a simple mathematical model of the A SNOM detected signal, this study explores the respective effects of the phase modulation depth, the wavelength and angle of the incident light, and the amplitude of the tip vibration on the signal contrast and signal intensity. In general, the results show that the background-noise decays as the order of the Bessel function increases and that higher-order harmonic frequencies yield an improved signal contrast. Additionally, it is found that incident light with a longer wavelength improves the signal contrast for a constant order of modulation frequency. The signal contrast can also be improved by reducing the incident angle of the incident light. Finally, it is demonstrated that sample stage scanning yields an improved imaging result. However, tip scanning provides a reasonable low-cost and faster solution in the smaller scan area. The analytical results presented in this study enable a better understanding of the complex detected signal in A-SNOM and provide insights into methods of improving the signal contrast of the A-SNOM measurement. PMID- 19550864 TI - Full-vectorial finite element method based eigenvalue algorithm for the analysis of 2D photonic crystals with arbitrary 3D anisotropy. AB - A full-vectorial finite element method based eigenvalue algorithm is developed to analyze the band structures of two-dimensional (2D) photonic crystals (PCs) with arbitray 3D anisotropy for in-planewave propagations, in which the simple transverse-electric (TE) or transverse-magnetic (TM) modes may not be clearly defined. By taking all the field components into consideration simultaneously without decoupling of the wave modes in 2D PCs into TE and TM modes, a full vectorial matrix eigenvalue equation, with the square of the wavenumber as the eigenvalue, is derived. We examine the convergence behaviors of this algorithm and analyze 2D PCs with arbitrary anisotropy using this algorithm to demonstrate its correctness and usefulness by explaining the numerical results theoretically. PMID- 19550865 TI - Optical coding scheme using optical interconnection for high sampling rate and high resolution photonic analog-to-digital conversion. AB - We propose and demonstrate an optical coding scheme using optical interconnection for a photonic analog-to-digital conversion. It allows us to convert a multi power level signal into a multiple-bit binary code so as to detect it in a bit parallel format by binary photodiode array. The proposed optical coding is executed after optical quantization using self-frequency shift. Optical interconnection based on a binary conversion table generates a multiple-bit binary code by appropriate allocation of a level identification signal which is provided as a result of optical quantization. Experimental results show that 8 levels analog pulses are converted into 3-bit parallel binary codes. PMID- 19550866 TI - Low voltage-defect quantum cascade laser with heterogeneous injector regions. AB - We demonstrate an In(0.635)Al(0.356)As/In(0.678)Ga(0.322)As strain compensated quantum cascade laser that employs heterogeneous injector regions for low voltage defect operation. The active core consists of interdigitated undoped and doped injectors followed by nominally identical wavelength optical transitions. The undoped injector regions are designed with reduced voltage defect while the doped injectors are of a more conventional design. The measured average voltage defect is less than 79 meV. At 80 K, a 2.3 mm long, back facet high reflectance coated laser has an emission wavelength of 4.7 mum and outputs 2.3 W pulsed power with a peak wall-plug efficiency of 19%. PMID- 19550867 TI - Parabolic pulse generation with active or passive dispersion decreasing optical fibers. AB - We experimentally demonstrate the possibility to generate parabolic pulses via a single dispersion decreasing optical fiber with normal dispersion. We numerically and experimentally investigate the influence of the dispersion profile, and we show that a hybrid configuration combining dispersion decrease and gain has several benefits on the parabolic generated pulses. PMID- 19550868 TI - Silicon photonic tunable optical dispersion compensator. AB - A silicon photonic tunable optical dispersion compensator (TODC) is demonstrated based on a series of 5 thermally tunable Mach- Zehnder interferometers. The TODC has a 2.8mm x 5.0 mm foot-print with continuously tunable dispersion from 0ps/nm to 2000ps/nm with a low tuning power of 80mW. This TODC is used to extend the reach of a 10Gb/s link from 85km to 150km. PMID- 19550869 TI - Realization of two Fourier-limited solid-state single-photon sources. AB - We demonstrate two solid-state sources of indistinguishable single photons. High resolution laser spectroscopy and optical microscopy were combined at T = 1.4 K to identify individual molecules in two independent microscopes. The Stark effect was exploited to shift the transition frequency of a given molecule and thus obtain single photon sources with perfect spectral overlap. Our experimental arrangement sets the ground for the realization of various quantum interference and information processing experiments. PMID- 19550870 TI - A refractometer based on a micro-slot in a fiber Bragg grating formed by chemically assisted femtosecond laser processing. AB - A liquid core waveguide as a refractometer is proposed. Microtunnels were created in standard optical fiber using tightly focused femtoscond laser inscription and chemical etching. A 1.2(h)x125(d) x500(l) mum micro-slot engraved along a fiber Bragg grating (FBG) was used to construct liquid core waveguide by filling the slot with index matching oils. The device was used to measure refractive index and sensitivity up to 10- 6/pm was obtained. PMID- 19550871 TI - Invariant resolution dynamic focus OCM based on liquid crystal lens. AB - A primary limitation of optical coherence microscopy is the lack of sufficient lateral resolution over a usable imaging volume for diagnostic applications, even with high-numerical aperture imaging optics. In this paper, we first motivate the benefit of refocusing at multiple depths in a highly scattering biological sample using optical coherence microscopy, which experimentally shows invariant 2.5 mum axial and 6.5 mum lateral resolution throughout the sample. We then present the optical system design of a hand-held probe with the advanced capability to dynamically focus with no moving parts to a depth of 2 mm in skin-equivalent tissue at 3 mum resolution throughout an 8 cubic millimeter imaging volume. The built-in dynamic focusing ability is investigated with an addressable liquid crystal lens embedded in a custom-designed optics optimized for a Ti:Sa pulsed broadband laser source of bandwidth 100nm centered at 800nm. The design was developed not only to account for refocusing into the tissue but also to minimize and compensate for the varying on-axis and off-axis optical aberrations that would be introduced throughout a 2 mm thick and 2 mm wide skin imaging volume. The MTF contrast functions and distortion plots at three different skin depths are presented. PMID- 19550872 TI - Electrically tunable fast light at THz bandwidth using cascaded semiconductor optical amplifiers. AB - Ultra fast non-linear processes are used to achieve an advance of 2 ps for a 600 fs pulse propagating through two SOAs in series. This corresponding 3.3-pulse advance is tuned continuously by changing the current applied to the devices. We propose an experimental scheme that uses a single SOA in a loop to emulate the propagation of pulse through multiple cascaded SOAs. PMID- 19550873 TI - Second-harmonic parametric scattering in ferroelectric crystals with disordered nonlinear domain structures. AB - We study the second-harmonic (SH) parametric processes in unpoled crystals of Strontium Barium Niobate (SBN) with disordered structures of ferroelectric domains. Such crystals allow for the simultaneous phase matching of several second-order nonlinear processes. We analyze the polarization properties of these parametric processes using two types of generation schemes: quasi-collinear SH generation and transverse SH generation. From our experimental data we determine the ratio of d(32) and d(33) components of the second order susceptibility tensor and also the statistical properties of the random structure of the SBN crystal. PMID- 19550874 TI - Coma measurement of projection optics in lithographic tools based on relative image displacements at multiple illumination settings. AB - In this paper, we propose a novel method for measuring the coma aberrations of lithographic projection optics based on relative image displacements at multiple illumination settings. The measurement accuracy of coma can be improved because the phase-shifting gratings are more sensitive to the aberrations than the binary gratings used in the TAMIS technique, and the impact of distortion on displacements of aerial image can be eliminated when the relative image displacements are measured. The PROLITH simulation results show that, the measurement accuracy of coma increases by more than 25% under conventional illumination, and the measurement accuracy of primary coma increases by more than 20% under annular illumination, compared with the TAMIS technique. PMID- 19550875 TI - Development of optical hyperlens for imaging below the diffraction limit. AB - We report here the design, fabrication and characterization of optical hyperlens that can image sub-diffraction-limited objects in the far field. The hyperlens is based on an artificial anisotropic metamaterial with carefully designed hyperbolic dispersion. We successfully designed and fabricated such a metamaterial hyperlens composed of curved silver/alumina multilayers. Experimental results demonstrate far-field imaging with resolution down to 125nm at 365nm working wavelength which is below the diffraction limit. PMID- 19550876 TI - High power and high energy ultrashort pulse generation with a frequency shifted feedback fiber laser. AB - The highest average power that has been achieved with a frequency-shifted feedback modelocked fiber laser is reported. Subpicosecond pulses with 40 kW peak power are obtained by this technique for the first time by using external pulse compression. The pulsing is self starting and environmentally stable. The measured pulse energy in modelocked operation is 120 nJ. The pulses could be compressed to 855 fs. The pulse energy was increased to 1muJ with controlled Q switched modelocking. PMID- 19550878 TI - An efficient Jacobian reduction method for diffuse optical image reconstruction. AB - Model based image reconstruction in Diffuse Optical Tomography relies on both the numerical accuracy of the forward model as well as the computational speed and efficiency of the inverse model. Most model based image reconstruction algorithms rely on Newton type inversion methods, whereby the inverse of a large Jacobian is approximated. In this work we present an efficient Jacobian reduction method which takes into account the total sensitivity of the imaging domain to the measured boundary data. It is shown using numerical and phantom data that by removing regions within the inverse model whose contribution to the measured data is less than 1%, it has no significant effect upon the estimated inverse problem, but does provide up to a 14 fold improvement in computational time. PMID- 19550877 TI - Time-domain upconversion measurements of group-velocity dispersion in quantum cascade lasers. AB - A time-resolved mid-infrared upconversion technique based on sum-frequency generation was applied to measure pulse propagation in lambda approximately 5.0 mum quantum cascade lasers operated in continuous wave at 30 K. The wavelength dependent propagation delay of femtosecond mid-infrared pulses was measured to determine the total group-velocity dispersion. The material and waveguide dispersion were calculated and their contributions to the total group-velocity dispersion were found to be relatively small and constant. The small-signal gain dispersion was estimated from a measurement of the electroluminescence spectrum without a laser cavity, and was found to be the largest component of the total GVD. A negative group-velocity dispersion of beta2 ( =d2beta/d omega2) approximately - 4.6x10-6 ps2/mum was observed at the peak emission wavelength, and good agreement was found for the measured and calculated pulse-broadening. PMID- 19550879 TI - Field trial of differential-phase-shift quantum key distribution using polarization independent frequency up-conversion detectors. AB - We report a field trial of differential phase shift quantum key distribution (QKD) using polarization independent frequency up-conversion detectors. A frequency up-conversion detector is a promising device for achieving a high key generation rate when combined with a high clock rate QKD system. However, its polarization dependence prevents it from being applied to practical QKD systems. In this paper, we employ a modified polarization diversity configuration to eliminate the polarization dependence. Applying this method, we performed a long term stability test using a 17.6-km installed fiber. We successfully demonstrated stable operation for 6 hours and achieved a sifted key generation rate of 120 kbps and an average quantum bit error rate of 3.14 %. The sifted key generation rate was not the estimated value but the effective value, which means that the sifted key was continuously generated at a rate of 120 kbps for 6 hours. PMID- 19550880 TI - Observation of Goos-Hanchen shifts in metallic reflection. AB - We report the first observation of the Goos-Hanchen shift of a light beam incident on a bare metal surface. This phenomenon is particularly interesting because the Goos-Hanchen shift for p polarized light in metals is negative and much bigger than the positive shift for s polarized light. The experimental result for the measured shifts as a function of the angle of incidence is in excellent agreement with theoretical predictions. In an energy-flux interpretation, our measurement shows the existence of a backward energy flow at the bare metal surface when this is excited by a p polarized beam of light. PMID- 19550881 TI - Fast computation of Lyot-style coronagraph propagation. AB - We present a new method for numerical propagation through Lyot-style coronagraphs using finite occulting masks. Standard methods for coronagraphic simulations involve Fast Fourier Transforms (FFT) of very large arrays, and computing power is an issue for the design and tolerancing of coronagraphs on segmented Extremely Large Telescopes (ELT) in order to handle both the speed and memory requirements. Our method combines a semi-analytical approach with non-FFT based Fourier transform algorithms. It enables both fast and memory-efficient computations without introducing any additional approximations. Typical speed improvements based on computation costs are of about ten to fifty for propagations from pupil to Lyot plane, with thirty to sixty times less memory needed. Our method makes it possible to perform numerical coronagraphic studies even in the case of ELTs using a contemporary commercial laptop computer, or any standard commercial workstation computer. PMID- 19550882 TI - SBS gain efficiency measurements and modeling in a 1714 mum(2) effective area LP(08) higher-order mode optical fiber. AB - The stimulated Brillouin scattering (SBS) gain efficiencies were measured in the LP(08) and LP(01) modes of a higher-order-mode optical fiber. Gain efficiencies C(B) of 0.0085 and 0.20 (m-W)(-1) were measured for the LP(08) and LP(01) modes at 1083 nm, respectively. C(B) is inversely proportional to the optical effective area Aeff and the same core-localized acoustic phonon seeds the SBS process in each case. An acoustic modal analysis and a distributed phenomenological model are presented to facilitate the data analysis and interpretation. The LP(08) mode exhibits a threshold powerlength product of 2.5 kW-m. PMID- 19550883 TI - Optical second harmonic generation imaging for visualizing in-plane electric field distribution. AB - The electric field distribution in electronic devices, particularly in the organic devices, was visualized by the optical second harmonic generation (SHG) imaging technique on the basis of electric field induced SHG (EFISHG). Two dimensional SHG images from organic field effect transistor using pentacene were taken with a cooled CCD camera, and the SHG images showed the electric field was successfully visualized with a resolution of 1 mum. The SHG imaging method provides us a novel technique for visualizing the electric field distribution in actual devices under device operation. PMID- 19550884 TI - Comparison of three-dimensional optical coherence tomography and high resolution photography for art conservation studies. AB - Gold punchwork and underdrawing in Renaissance panel paintings are analyzed using both three-dimensional swept source / Fourier domain optical coherence tomography (3D-OCT) and high resolution digital photography. 3D-OCT can generate en face images with micrometer-scale resolutions at arbitrary sectioning depths, rejecting out-of-plane light by coherence gating. Therefore 3D-OCT is well suited for analyzing artwork where a surface layer obscures details of interest. 3D-OCT also enables cross-sectional imaging and quantitative measurement of 3D features such as punch depth, which is beneficial for analyzing the tools and techniques used to create works of art. High volumetric imaging speeds are enabled by the use of a Fourier domain mode locked (FDML) laser as the 3D-OCT light source. High resolution infrared (IR) digital photography is shown to be particularly useful for the analysis of underdrawing, where the materials used for the underdrawing and paint layers have significantly different IR absrption properties. In general, 3D-OCT provides a more flexible and comprehensive analysis of artwork than high resolution photography, but also requires more complex instrumentation and data analysis. PMID- 19550885 TI - Dynamic spectroscopic ellipsometry determination of nanostructural changes in plasmonic silver films. AB - Dynamic in situ spectroscopic ellipsometry is used to probe post-deposition nano structural changes in silver films at room temperature in the pre- and post coalescence stages of Volmer-Weber growth. In the island growth phase the Maxwell Garnett theory is used to determine structural changes in the island film. Changes in the plasmon resonance frequency indicate an increased distance between islands which explain pre-coalescence resistivity changes. Post-coalescence changes in the resistivity are determined to be due to grain growth. A reduction in film thickness of 0.2 - 0.3 nm is also observed. The results are used to evaluate recent competing theories based on in situ stress measurements. PMID- 19550886 TI - MLSE receiver tolerance to all-order polarization mode dispersion. AB - We experimentally characterize a maximum likelihood sequence estimation (MSLE) based receiver's tolerance to first- and all-order polarization mode dispersion (PMD). We show that the response of the MLSE receiver to first-order PMD can be characterized in two ways depending on the differential group delay (DGD). In addition we show that first-order PMD-induced system penalties dominate those from high-order PMD. High-order PMD induces a large system penalty only when the first-order penalty is small, or the DGD exceeds a bit period. PMID- 19550887 TI - 2-D PSTD Simulation of optical phase conjugation for turbidity suppression. AB - Turbidity Suppression via Optical Phase Conjugation (TS-OPC) is an optical phenomenon that uses the back propagation nature of optical phase conjugate light field to undo the effect of tissue scattering. We use the computationally efficient and accurate pseudospectral time-domain (PSTD) simulation method to study this phenomenon; a key adaptation is the volumetric inversion of the optical wavefront E-field as a means for simulating a phase conjugate mirror. We simulate a number of scenarios and demonstrate that TS-OPC deteriorates with increased scattering in the medium, or increased mismatch between the random medium and the phase conjugate wave during reconstruction. PMID- 19550888 TI - Passively mode-locked and cavity-dumped Yb:KY(WO(4))(2) oscillator with positive dispersion. AB - We demonstrate, what is to our knowledge the first passively mode-locked Ytterbium based solid state high energy laser oscillator operated in the positive dispersion regime. Compared to solitary mode-locking the pulse energy can be increased with even broader spectral bandwidth. With high speed cavity dumping the laser generates 2 muJ-pulses at a 1 MHz repetition rate. The chirped output pulses are compressible down to 420 fs. PMID- 19550889 TI - Mode-locked X-wave lasers. AB - We theoretically demonstrate X-waves as global attractors that enable mode locking of a laser cavity operating in the normal dispersion regime. This result is based upon a fully comprehensive physical model of the laser cavity, where the nonlinear discrete diffraction dynamics of a waveguide array mediates the spontaneous periodic generation of spatio-temporal X-waves. PMID- 19550890 TI - Calculation of spherical red blood cell deformation in a dual-beam optical stretcher. AB - We present a numerical method based on the linear elastic membrane theory to compute the morphological deformation of a spherical cell from the photonics stress distribution over the cellular membrane. The method is applied to fit the experimental data for deformation of a spherical human red blood cell trapped and stretched in a fiber-optical dual-beam trap with a single fitting parameter Eh where E is the Young's modulus of elasticity and h is the thickness of the cell membrane. We obtained Eh = (20+/-2)muNm(-1) which is comparable to results reported earlier. This numerical method can be applied in general experimental conditions. PMID- 19550891 TI - Grating-free Raman laser using highly nonlinear photonic crystal fiber. AB - We demonstrate a Raman laser made from a grating-free highly-nonlinear photonic crystal fiber. The laser threshold power is lower than 600 mW and laser power characteristics recorded in experiments are accurately described from the usual simplest model dealing only with stationary evolutions of total optical powers [J. Opt. Soc. Am. 69, 803-807 (1979)]. In our theoretical treatment, reflectivity coefficients are fixed parameters, in strong contrast with procedures usually implemented to describe Raman fiber lasers made with fiber Bragg gratings. Experimental investigations of the spectral properties of our grating-free Raman fiber laser evidence that the shape of the Stokes power spectrum remains remarkably Gaussian whatever the incident pump power. Increasing the incident pump power induces a drift of the Stokes wavelength together with a broadening of the Stokes optical spectrum. Investigations on the role of light polarization on laser characteristics show that our grating-free Raman fiber laser behaves as a Raman laser made with a standard polarization maintaining fiber. PMID- 19550892 TI - Optical information transfer between two light channels in a Pr(3+):Y(2)SiO(5)crystal. AB - We experimentally demonstrate light storage and release in a four-level double lambda atomic system of a Pr (3+):Y(2)SiO(5) crystal. Based on the technique of light storage, we realize optical information transfer between two light channels. The coherent optical information of a probe pulse stored in the crystal can be selectively released into two different light channels by varying the frequency and propagation direction of the switch-on control field. PMID- 19550893 TI - Defocused imaging of second harmonic generation from a single nanocrystal. AB - We demonstrate the direct imaging of the second harmonic generation radiation from a single nonlinear nanocrystal using defocused nonlinear microscopy. This technique allows the retrieval of complete information on the 3D orientation of a nanocrystal as well as possible deviations from its purely crystalline nature, in a simple experimental implementation. The obtained images are modeled by calculation of the radiation diagram from a nonlinear dipole that accounts for the excitation beam, the crystal symmetry and the particle size. Experimental demonstrations are performed on Potassium Titanyl Phosphase (KTP) nanocrystals. The shape and structure of the radiation images show a strong dependence on both crystal orientation and field polarization state, as expected by the specific nonlinear coherent coupling between the induced dipole and the excitation field polarization state. PMID- 19550894 TI - In-situ femtosecond laser pulse characterization and compression during micromachining. AB - We report on phase measurements and adaptive phase distortion compensation of femtosecond pulses using multiphoton intrapulse interference phase scan (MIIPS) based on second harmonic generation in the plasma generated on the surface of silicon and metals. PMID- 19550895 TI - Double image encryption based on random phase encoding in the fractional Fourier domain. AB - A novel image encryption method is proposed by utilizing random phase encoding in the fractional Fourier domain to encrypt two images into one encrypted image with stationary white distribution. By applying the correct keys which consist of the fractional orders, the random phase masks and the pixel scrambling operator, the two primary images can be recovered without cross-talk. The decryption process is robust against the loss of data. The phase-based image with a larger key space is more sensitive to keys and disturbances than the amplitude-based image. The pixel scrambling operation improves the quality of the decrypted image when noise perturbation occurs. The novel approach is verified by simulations. PMID- 19550896 TI - Color information processing (coding and synthesis) with fractional Fourier transforms and digital holography. AB - In this paper, we propose a new method for color image coding and synthesis based on fractional Fourier transforms and wavelength multiplexing with digital holography. A color image is divided into three channels and each channel, in which the information is encrypted with different wavelength, fractional orders and random phase masks, is independently encrypted or synthesized. The system parameters are additional keys and this method would improve the security of information encryption. The images are fused or subtracted by phase shifting technique. The possible optical implementations for color image encryption and synthesis are also proposed with some simulation results that show the possibility of the proposed idea. PMID- 19550897 TI - Ultra-High Q/V Fabry-Perot microcavity on SOI substrate. AB - We experimentally demonstrate an ultra high Q/V nanocavity on SOI substrate. The design is based on modal adaptation within the cavity and allows to measure a quality factor of 58.000 for a modal volume of 0.6(lambda/n)(3) . This record Q/V value of 10(5) achieved for a structure standing on a physical substrate, rather than on membrane, is in very good agreement with theoretical predictions also shown. Based on these experimental results, we show that further refinements of the cavity design could lead to Q/V ratios close to 10(6). PMID- 19550898 TI - Direct diode-pumped laser operation of Cr(3+)- doped LiInGeO(4) crystals. AB - Quasi-continuous wave (cw) laser action has been demonstrated by direct diode pumping of a new extremely broadband Cr(3+)-doped crystal. In contrast to previous works, where large-frame pump lasers have been employed, we have shown that low-power direct diode pumping of LiInGeO(4) is feasible, opening up the way for realizing compact and efficient laser sources for telecommunication applications. PMID- 19550899 TI - Optimization of multiple filamentation of femtosecond laser pulses in air using a pinhole. AB - The robustness and prolongation of multiple filamentation (MF) for femtosecond laser propagation in air are investigated experimentally and numerically. It is shown that the number, pattern, propagation distance, and spatial stability of MF can be controlled by a variable-aperture on-axis pinhole. The random MF pattern can be optimized to a deterministic pattern. In our numerical simulations, we configured double filaments to principlly simulate the experimental MF interactions. It is experimentally and numerically demonstrated that the pinhole can reduce the modulational instability of MF and is favorable for a more stable MF evolution. PMID- 19550900 TI - Mode profile dispersion in the generalised nonlinear Schrodinger equation. AB - The formulation of Schrodinger-like equations for nonlinear pulse propagation in a single-mode microstructured optical fiber with a strongly frequency-dependent guided-mode profile is investigated.A correct account of mode profile dispersion in general necessiates a generalization of the effective area concept commonly used in the generalized nonlinear Schrodinger equation (GNLSE). A numerical scheme to this end is developed, and applied to a solid-core photonic bandgap fiber as a test case. It is further shown, that a simple reformulation of the GNLSE, expressed only in terms of the traditional frequency-dependent effective area, yields a good agreement with the more complete theory. PMID- 19550901 TI - Broadband ultrafast spectroscopy using a photonic crystal fiber: application to the photophysics of malachite green. AB - Femtosecond single-pulsed supercontinua (SC) are produced in a short sub-cm piece of photonic crystal fiber. The SC span from 450 nm to more than 1.1 mum with 1-nJ energy injection. UV light down to 340 nm is observed with increased injection power. Using such a single-pulsed SC we implemented a compact transient absorption spectrometer with broadband detection and 150-fs FWHM time resolution to monitor the ultrafast dynamics of the electronic states of malachite green in ethanol excited to the S(2) state. The full spectral evolution is observed from 450 nm to 1050 nm, with high sensitivity and a signal-to-noise ratio as high as 1000. PMID- 19550902 TI - Optimization and characterization of a structured illumination microscope. AB - Structured illumination microscopy provides a simple and cheap mean to obtain optical sections of a sample. It can be implemented easily on a regular fluorescent microscope and is a scanning free alternative to confocal microscopy.We have analyzed theoretically the performances of the technique in terms of sectioning strength, resolution enhancement along the optical axis, and signal to background as a function of the objective used and the grid's characteristics (pitch and contrast). We show that under optimized conditions, the axial resolution can be improved by a factor of 1.5 in comparison with an epifluorescence microscope, and that optical cuts with a thickness of less than 400nm can be obtained with a 1.4 numerical aperture objective. We modified the original grid in-step modulation pattern and used a sinusoidal modulation for the grid displacement. Optical sections are computed by combining four images acquired during one modulation period. This algorithm is very stable even for modulations at high frequencies. The speed of the acquisition is thus only limited by the performance of the detector and the signal/background ratio of the sample. Finally, we compared our technique to commercial setups: a confocal microscope, a Spinning Disk Microscope and a Zeiss Apotome. PMID- 19550904 TI - Enhanced nonlinear optics in photonic-crystal microcavities. AB -

Focus Serial: Frontiers of Nonlinear Optics

Nonlinear photonic-crystal microresonators offer unique fundamental ways of enhancing a variety of nonlinear optical processes. This enhancement improves the performance of nonlinear optical devices to such an extent that their corresponding operation powers and switching times are suitable for their implementation in realistic ultrafast integrated optical devices. Here, we review three different nonlinear optical phenomena that can be strongly enhanced in photonic crystal microcavities. First, we discuss a system in which this enhancement has been successfully demonstrated both theoretically and experimentally, namely, a photonic crystal cavity showing optical bistability properties. In this part, we also present the physical basis for this dramatic improvement with respect to the case of traditional nonlinear devices based on nonlinear Fabry-Perot etalons. Secondly, we show how nonlinear photonic crystal cavities can be also used to obtain complete second-harmonic frequency conversion at very low input powers. Finally, we demonstrate that the nonlinear susceptibility of materials can be strongly modified via the so-called Purcell effect, present in the resonant cavities under study. PMID- 19550903 TI - In vivo functional imaging of human cone photoreceptors. AB - We evaluate a novel non-invasive optical technique for observing fast physiological processes, in particular phototransduction, in single photoreceptor cells in the living human eye. The method takes advantage of the interference of multiple reflections within the outer segments (OS) of cones. This self interference phenomenon is highly sensitive to phase changes such as those caused by variations in refractive index and scatter within the photoreceptor cell. A high-speed (192 Hz) flood-illumination retina camera equipped with adaptive optics (AO) is used to observe individual photoreceptors, and to monitor changes in their reflectance in response to visible stimuli ("scintillation"). AO and high frame rates are necessary for resolving individual cones and their fast temporal dynamics, respectively. Scintillation initiates within 5 to 10 ms after the onset of the stimulus flash, lasts 300 to 400 ms, is observed at visible and near-infrared (NIR) wavelengths, and is highly sensitive to the coherence length of the imaging light source. To our knowledge this is the first demonstration of in vivo optical imaging of the fast physiological processes that accompany phototransduction in individual photoreceptors. PMID- 19550905 TI - Liquid crystal Adaptive Optics Visual Simulator: Application to testing and design of ophthalmic optical elements. AB - The concept of Adaptive Optics Visual Simulation applies to the use of an Adaptive Optics system to manipulate ocular aberrations in order to perform visual testing through a modified optics. It can be of interest both to study the visual system and to design new ophthalmic optical elements. In this work, we describe an apparatus based on a liquid crystal programmable phase modulator and explore its capabilities as a tool in the early stages of the design of ophthalmic optical elements with increased depth of field for presbyopic subjects. To illustrate the potential of the instrument, we analyze the performance of two phase profiles obtained by a hybrid optimization procedure. The liquid crystal Adaptive Optics Visual Simulator can be used to experimentally record the point spread function for different vergences in order to objectively measure depth of focus, to perform different psychophysical experiments through the phase profile in order to measure its impact on visual performance, and to study the interaction with the eye's particular aberrations. This approach could save several steps in current procedures of ophthalmic optical design and eventually lead to improved solutions. PMID- 19550906 TI - Photon-counting passive 3D image sensing for reconstruction and recognition of partially occluded objects. AB - In this paper, we discuss the reconstruction and the recognition of partially occluded objects using photon counting integral imaging (II). Irradiance scenes are numerically reconstructed for the reference target in three-dimensional (3D) space. Photon counting scenes are estimated for unknown input objects using maximum likelihood estimation (MLE) of Poisson parameter. We propose nonlinear matched filtering in 3D space to recognize partially occluded targets. The recognition performance is substantially improved from the nonlinear matched filtering of elemental images without 3D reconstruction. The discrimination capability is analyzed in terms of Fisher ratio (FR) and receiver operating characteristic (ROC) curves. PMID- 19550907 TI - Improved performance in coaxial holographic data recording. AB - We describe a coaxial holographic recording system for achieving high recording density. We implement several techniques, such as an objective lens with high numerical aperture (NA), high capacity page data format, a random binary phase mask, and an optical noise reduction element. Our system successfully realizes a hologram recording/retrieving at a low diffraction efficiency less than 2.0 x 10( 3) and achieves a raw data density of 180 Gbit/in.(2), thus demonstrating the potential of a coaxial holographic system for high-density optical storage systems. PMID- 19550908 TI - Experimental demonstration of a wafer-level flexible probe for optical waveguide testing. AB - A flexible optical probe that accomplishes wafer-level directional coupling of light into optical waveguides is investigated theoretically and experimentally. Simulated results indicate high coupling efficiencies in excess of 80% for a range of parameters. Probe fabrication was implemented using SU8 as flexible waveguide material. Coupling of light from flexible probe to an S-shaped test waveguide demonstrated 11% efficiency compared to direct butt coupling. These results may lead to increased yield, shorter development cycles and overall savings in PLC packaging costs. PMID- 19550909 TI - Ring surface waves in thermal nonlinear media. AB - We address ring-shaped surface waves supported by defocusing thermal media with circular cross-section. Such waves exist because of the balance between repulsion from the interface and deflection of light from the bulk medium due to defocusing nonlocal nonlinearity. The properties of such surface waves are determined by the geometry of the sample. Nodeless ring surface waves are stable for all values of their winding number, while surface waves with a small number of azimuthal nodes can be metastable . PMID- 19550910 TI - Analysis of Smith-Purcell radiation in optical region. AB - Smith-Purcell radiation (SPR), emitted when an electron beam is traveling above a metallic grating, has attracted a lot of attention as a source of electromagnetic (EM) radiation in the millimeter to visible spectrum. We conducted a theoretical investigation of SPR in the optical region using a two-dimensional finite difference time-domain (FDTD) method. The permittivity of metal was represented using the Drude model. During the simulation, we observed three types of EM radiations when an electron bunch passes above a metal grating. We think these three types of EM radiation were basic SPR, original surface plasmon polariton (SPP), and mimic-SPP, caused by the periodic grating structure. Our observations were in accordance with analytical models of original SPP and mimic-SPP EM radiation. PMID- 19550911 TI - Models of dielectric response in disordered solids. AB - Two dispersion models of disordered solids, one parameterizing density of states (PDOS) and the other parameterizing joint density of states (PJDOS), are presented. Using these models, not only the complex dielectric function of the materials, but also some information about their electronic structure can be obtained. The numerical integration is necessary in the PDOS model. If analytical expressions are required the presented PJDOS model is, for some materials, a suitable option still providing information about the electronic structure of the material. It is demonstrated that the PDOS model can be successfully applied to a wide variety of materials. In this paper, its application to diamond-like carbon (DLC), a-Si and SiO2-like materials are discussed in detail. Unlike the PDOS model, the presented PJDOS model represents a special case of parameterization that can be applied to limited types of materials, for example DLC, ultrananocrystalline diamond (UNCD) and SiO2-like. PMID- 19550912 TI - Design of resonant microcavities: application to optical gyroscopes. AB - We study theoretically and numerically the effect of rotation on resonant frequencies of microcavities in a rotating frame of reference. Cavity rotation causes the shifts of the resonant frequencies proportional to the rotation rate if it is larger than a certain value. Below the value, a region of rotation rate exists where there is no resulting the frequency shifts proportional to the rotation rate. We show that designing cavity symmetry as C(nv) (n >/= 3) can eliminate this region. PMID- 19550913 TI - Enhanced viewing-angle integral imaging by multiple-axis telecentric relay system. AB - One of the main limitations of integral imaging is the narrow viewing angle. This drawback comes from the limited field of view of microlenses during the pickup and display. We propose a novel all-optical technique which allows the substantial increase of the field of view of any microlens and therefore of the viewing angle of integral-imaging displays. PMID- 19550914 TI - In vivo monitoring of blood oxygenation in large veins with a triple-wavelength optoacoustic system. AB - A noninvasive optoacoustic technique could be a clinically useful alternative to existing, invasive methods for cerebral oxygenation monitoring. Recently we proposed to use an optoacoustic technique for monitoring cerebral blood oxygenation by probing large cerebral and neck veins including the superior sagittal sinus and the internal jugular vein. In these studies we used a multi wavelength optoacoustic system with a nanosecond optical parametric oscillator as a light source and a custom-made optoacoustic probe for the measurement of the optoacoustic signals in vivo from the area of the sheep neck overlying the external jugular vein, which is similar in diameter and depth to the human internal jugular vein. Optoacoustic signals induced in venous blood were measured with high resolution despite the presence of a thick layer of tissues (up to 10 mm) between the external jugular vein and the optoacoustic probe. Three wavelengths were chosen to provide accurate and stable measurements of blood oxygenation: signals at 700 nm and 1064 nm demonstrated high correlation with actual oxygenation measured invasively with CO-Oximeter ("gold standard"), while the signal at 800 nm (isosbestic point) was independent of blood oxygenation and was used for calibration. PMID- 19550915 TI - Selective coating of holes in microstructured optical fiber and its application to in-fiber absorptive polarizers. AB - An interesting feature of microstructured optical fibers (MOFs) is that their properties can be adjusted by filling or coating of the holes. Some applications require selective filling or coating, which has proved experimentally demanding. We demonstrate selective coating of MOFs with metal and use it to fabricate an in fiber absorptive polarizer. PMID- 19550916 TI - Segmented grown Yb:KY(WO(4))(2)/KY(WO(4))2(2) for use in continuous-wave and mode locked lasers. AB - Segmented growth of monoclinic Yb:KY(WO(4))(2) on KY(WO(4))2(2) substrates was successfully implemented and its excellent laser performance demonstrated. High slope efficiencies up to 80% and an output power of 375 mW were achieved under Ti:sapphire laser pumping in the continuous-wave regime. In the passively mode locked regime, pulses as short as 99 fs with an average output power of 69 mW were obtained. PMID- 19550917 TI - Encapsulation of low-refractive-index SiO(2) nanorods by Al(2)O(3) with atomic layer deposition. AB - Thin films composed of SiO(2) nanorods or nanoporous SiO(2) (np- SiO(2)) are attractive for use as a low refractive index material in various types of optical coatings. However, the material properties of these films are unstable because of the high porosity of the films. This is particularly apparent in dry versus humid atmospheres where both the refractive index and coefficient of thermal expansion (CTE) vary dramatically. In this article, we demonstrate that np-SiO(2) can be encapsulated by depositing Al(2)O(3) with Atomic Layer Deposition (ALD), stabilizing these properties. In addition, this encapsulation ability is demonstrated successfully in a 4-pair distributed Bragg reflector (DBR) design. It is hoped that this technique will be useful in patterning specific regions of a film for optical and mechanical stability while other portions are ambient interactive for sensing. PMID- 19550918 TI - Observation of Q-switching and mode-locking in two-section InAs/InP (100) quantum dot lasers around 1.55 mum. AB - First observation of passive mode-locking in two-section quantum-dot lasers operating at wavelengths around 1.55 mum is reported. Pulse generation at 4.6 GHz from a 9 mm long device is verified by background-free autocorrelation, RF spectra and real-time oscilloscope traces. The output pulses are stretched in time and heavily up-chirped with a value of 20 ps/nm, contrary to what is normally observed in passively mode-locked semiconductor lasers. The complete output spectrum is shown to be coherent over 10 nm. From a 7 mm long device Q switching is observed over a large operating regime. The lasers have been realized using a fabrication technology that is compatible with further photonic integration. This makes the laser a promising candidate for e.g. a mode-comb generator in a complex photonic chip. PMID- 19550919 TI - Soliton fission management by dispersion oscillating fiber. AB - We report the experimental observation of the fission of picosecond solitons in a fiber with sine-wave variation of the core diameter along the longitudinal direction of propagation. The experimental pulse dynamics is reproduced by numerical simulations. The fission of high-intensity solitons caused by both the variation of the fiber dispersion and stimulated Raman scattering is demonstrated. The number of output pulses and their frequencies can be managed by periodical modulation of the fiber dispersion even under the strong effect of the Raman scattering. PMID- 19550920 TI - GaP waveguide emitters for high power broadband THz generation pumped by Yb-doped fiber lasers. AB - We demonstrate the generation of broadband THz pulses by optical rectification in GaP waveguides pumped by high power Yb-doped fiber amplifiers. The dispersion of the GaP emitter can be controlled via the geometry of the waveguide; the peak frequency of the emitted THz radiation is tuned by varying the waveguide cross section. Most importantly, the use of a waveguide for the THz emission increases the coherent buildup length of the THz pulses and offers scalability to higher power; this was investigated by pumping a GaP waveguide emitter with a high power Yb-doped fiber laser system. A 25-MHz-repetition-rate pulse train of THz radiation with 120 muW average power was achieved using 14 W optical power, which represents the highest average power for a broadband THz source pumped by fiber lasers to date. PMID- 19550921 TI - Discrete wavelength tuning characteristics of a single-frequency fiber laser with dual-wavelength external frequency-stable light injection. AB - We investigate discrete wavelength tuning characteristics of a single-frequency fiber laser locked to either of two external lights. Frequency locking is achieved by the cooperatively induced spatial-hole burning (SHB) of a saturable absorber in the laser cavity. We show that lasing frequency is well locked to either of the two external lights when the lasing wavelength of the fiber laser is adjusted to the corresponding wavelength of the external light by tuning the bandpass filter in the laser cavity. The locked frequency stability of the fiber laser is as high as that of the employed external light source. PMID- 19550922 TI - Speckle mechanism in holographic optical imaging. AB - Speckle imaging was investigated by using dynamic holography and photorefractive AlGaAs/GaAs multiple quantum wells in a holographic optical imaging system. We showed that the speckle contrast depends on holographic fringes and the photorefractive effect. We further demonstrated that a moving grating technique can be used to suppress the random speckle. PMID- 19550923 TI - Rytov theory for Helmholtz-Gauss beams in turbulent atmosphere. AB - The Rytov theory for the propagation of Helmholtz-Gauss (HzG) beams in turbulent atmosphere is presented. We derive expressions for the first and second-order normalized Born approximations, the second-order moments, and the transverse intensity pattern of the HzG beams at any arbitrary propagation distance. The general formulation is applied to study the propagation of several special cases of the HzG beams, in particular, the Bessel-Gauss and Mathieu-Gauss beams and their pure nondiffracting counterparts, the Bessel and Mathieu beams. For numerical purposes, we assume the standard Kolmogorov distribution to model the power spectrum of the atmospheric fluctuations. PMID- 19550924 TI - Polarisation-mode coupling in (100)-cut Nd:YAG. AB - We investigate the polarisation-mode dynamics and Lamb's mode coupling constant for orthogonally polarised laser states in a dual-mode (100)-cut Nd:YAG laser with feedback, and compare with an anisotropic rate equation model. The anisotropic (100)-cut Nd:YAG exhibits thermally-induced depolarisation and polarisation-mode coupling dependent on the pump polarisation, crystal angle and laser polarisation directions. Here, the links between the depolarisation and polarisation-mode coupling are discussed with reference to a rate equation model which includes gain anisotropy in a quasi-isotropic laser cavity. PMID- 19550925 TI - 13.5 nm EUV generation from tin-doped droplets using a fiber laser. AB - A comprehensive study of the spectral and Mo-Si mirror inband EUV emission from tin-doped droplet laser plasma targets irradiated with a single 1064 nm beam from an Yb:doped fiber laser is reported.With pre-pulse enhancement, in-band conversion efficiency of approximately 2.1% is measured for laser irradiance intensities near 8 x 10(10) W/cm(2). This is the first study to be reported that uses a high-power, high repetition rate fiber laser with the high repetition rate droplet targets where EUV generation from plasmas is measured. PMID- 19550926 TI - Dual chirped optical pulses from a phase-modulated laser. AB - Dual chirped optical pulse is reported by use of a phase-modulated laser. When a number of sidebands around the carrier frequency in a phase-modulated laser spectrum are spectrally carved, short optical pulse trains with dual chirp states are obtained. It is discussed that the pulse duration and the linear frequency chirp parameter can be found in closed form expressions given as a function of the modulation index and the modulation frequency when the phase modulation function of the dual-chirped pulse trains does not deviate severely from the modulation function applied to the phase modulator by spectral filtering effects. In experiments, optical pulse trains at 5 GHz repetition rate were generated with the pulse duration of 29.4 ps when frequency-modulated fiber laser signals with a 2.5 GHz repetition rate passed through a Gaussian-shaped optical band-pass filter (OBF) with a full width at half maximum (FWHM) 0.3 nm and a transmission peak at the carrier frequency of the frequency-modulated laser spectrum. Measured pulse duration was in good agreement with the analytical prediction value 28.2 ps and the numerical result 27.7 ps. PMID- 19550927 TI - An optical tweezer actuated, nanoaperture-grid based Optofluidic Microscope implementation method. AB - We report a novel grid based Optofluidic Microscope (OFM) method where a closely spaced 2D grid of nanoapertures (diameter = 100 nm, separation = 2.5 mum) provided patterned illumination. We achieved a one-to-one mapping of the light transmissions through the nanoapertures onto a high-speed CCD camera. By optically tweezing a targeted sample across the grid in a controlled fashion and recording the time varying light reception from the nanoapertures, we were able to generate high-resolution images of the sample. The achievable resolution limit of the prototype was ~ 110 nm (Sparrow's criterion) under optimal conditions. We demonstrated the technique by imaging polystyrene beads and pollen spores. PMID- 19550928 TI - Efficient waveguide-integrated tunnel junction detectors at 1.6 mum. AB - Near-infrared detectors based on metal-insulator-metal tunnel junctions integrated with planarized silicon nanowire waveguides are presented, which we believe to be the first of their kind. The junction is coupled to the waveguide via a thin-film metal antenna feeding a plasmonic travelling wave structure that includes the tunnel junction. These devices are inherently broadband; the design presented here operates throughout the 1500-1700 nm region. Careful design of the antenna and travelling wave region substantially eliminates losses due to poor mode matching and RC rolloff, allowing efficient operation. The antennas are made from multilayer stacks of gold and nickel, and the active devices are Ni-NiO-Ni edge junctions. The waveguides are made via shallow trench isolation technology, resulting in a planar oxide surface with the waveguides buried a few nanometres beneath.The antennas are fabricated using directional deposition through a suspended Ge shadow mask, using a single level of electron-beam lithography. The waveguides are patterned with conventional 248-nm optical lithography and reactive-ion etching, then planarized using shallow-trench isolation technology. We also present measurements showing overall quantum efficiencies of 6% (responsivity 0.08 A/W at 1.605 mum), thus demonstrating that the previously very low overall quantum efficiencies reported for antenna-coupled tunnel junction devices are due to poor electromagnetic coupling and poor choices of antenna metal, not to any inherent limitations of the technology. PMID- 19550929 TI - Spectrally balanced detection for optical frequency domain imaging. AB - In optical frequency domain imaging (OFDI) or swept-source optical coherence tomography, balanced detection is required to suppress relative intensity noise (RIN). A regular implementation of balanced detection by combining reference and sample arm signal in a 50/50 coupler and detecting the differential output with a balanced receiver is however, not perfect. Since the splitting ratio of the 50/50 coupler is wavelength dependent, RIN is not optimally canceled at the edges of the wavelength sweep. The splitting ratio has a nearly linear shift of 0.4% per nanometer. This brings as much as +/-12% deviation at the margins of wavelength swept range centered at 1060nm. We demonstrate a RIN suppression of 33dB by spectrally corrected balanced detection, 11dB more that regular balanced detection. PMID- 19550932 TI - Backscattering of light from disk-like particles with aperiodic angular fine structure. AB - Recent computations of the backscattering cross section (sigmab) of randomly oriented disk-like particles (refractive index, 1.20) with small-scale periodic angular internal structure, have been repeated for similarly sized particles, but with the periodic structure replaced by an aperiodic structure. The latter is formed by randomly perturbing a periodic structure. Although sigmab for individual realizations of an aperiodic disk can differ significantly from that of its periodic counterpart, averaging over several realizations brings the two into confluence, unless the aperiodicity is too large. These computations suggest that using disks with perfectly periodic (as opposed to quasi-periodic) fine structure for modeling the backscattering of detached coccoliths from E. Huxleyi is justified. PMID- 19550930 TI - Linear 3D reconstruction of time-domain diffuse optical imaging differential data: improved depth localization and lateral resolution. AB - We present 3D linear reconstructions of time-domain (TD) diffuse optical imaging differential data. We first compute the sensitivity matrix at different delay gates within the diffusion approximation for a homogeneous semi-infinite medium. The matrix is then inverted using spatially varying regularization. The performances of the method and the influence of a number of parameters are evaluated with simulated data and compared to continuous-wave (CW) imaging. In addition to the expected depth resolution provided by TD, we show improved lateral resolution and localization. The method is then applied to reconstructing phantom data consisting of an absorbing inclusion located at different depths within a scattering medium. PMID- 19550931 TI - Subcellular-resolution molecular imaging within living tissue by fiber microendoscopy. AB - Conventional histopathology involves sampling, sectioning and staining of tissue specimens prior to microscopic evaluation, and provides diagnostic information at a single location and point in time. In vivo microscopy and molecular-targeted optical labeling are two rapidly developing fields, which together have the potential to provide anatomical and functional indications of disease by staining and imaging tissue in situ. To address the need for high-resolution imaging instrumentation, we have developed a compact, robust, and inexpensive fiber-optic microendoscopy system based around wide-field LED illumination, a flexible 1 mm diameter fiber-optic bundle, and a color CCD camera. We demonstrate the sub cellular resolution imaging capabilities of the system through a series of experiments, beginning with simultaneous imaging of three different cancer cell lines in culture, each targeted with a distinct fluorescent label. We used the narrow diameter probe to access subcutaneous tumors in an in vivo murine model, allowing direct comparison of microendoscopy images with macroscopic images and histopathology. A surgically resected tissue specimen from the human oral cavity was imaged across the clinical margin, demonstrating qualitative and quantitative distinction between normal and cancerous tissue based on sub-cellular image features. Finally, the fiber-optic microendoscope was used on topically-stained normal human oral mucosa in vivo, resolving epithelial cell nuclei and membranes in real-time fluorescence images. Our results demonstrate that this imaging system can potentially complement conventional diagnostic techniques, and support efforts to translate emerging molecular-diagnostic and therapeutic agents into clinical use. PMID- 19550933 TI - Fresnel diffraction effects in Fourier-transform arrayed waveguide grating spectrometer. AB - We present an analysis of Fourier-transform arrayed waveguide gratings in the Fresnel diffraction regime. We report a distinct spatial modulation of the interference pattern referred to as the Moire-Talbot effect. The effect and its influence in a FT AWG device is explained by deriving an original analytical expression for the modulated field, and is also confirmed by numerical simulations using the angular spectrum method to solve the Fresnel diffraction integral. We illustrate the retrieval of spectral information in a waveguide Fourier-transform spectrometer in the presence of the Moire-Talbot effect. The simulated device comprises two interleaved waveguide arrays each with 180 waveguides and the interference order of 40. It is designed with a Rayleigh spectral resolution of 0.1 nm and 8 nm bandwidth at wavelength lambda approximately 1.5 mum. We also demonstrate by numerical simulations that the spectrometer crosstalk is reduced from -20 dB to -40 dB by Gaussian apodization. PMID- 19550934 TI - Experimental demonstration of interference avoidance protocol (transmission scheduling) in O-CDMA networks. AB - We experimentally demonstrate a transmission scheduling algorithm to avoid congestion collapse in O-CDMA networks. Our result shows that transmission scheduling increases the performance of the system by orders of magnitude. PMID- 19550935 TI - Towards the short-wavelength limit lasing at 1450 nm over 4I13/2 ? 4I15/2 transition in silica-based erbium-doped fiber. AB - The transition rate of the stimulated emission at the higher energy levels of the excited states in a silica-based erbium-doped fiber can be enhanced by introducing fundamental-mode cutoff filtering mechanism. The electrons excited by optical pumping can more occupy the higher energy levels of the excited states when the transition rate for the lower energy levels (longer wavelengths) of the excited states is substantially suppressed. The achieved lasing wavelength can thus be moving toward the shorter wavelengths of the gain bandwidth. The laser transition between 4I13/2 ? 4I15/2 multiplets of the silica-based erbium-doped fiber is known to emit fluorescence with the shortest wavelength around 1450 nm. We, for the first time, experimentally demonstrate a widely tunable fiber laser at the wavelength very close to 1450 nm by using a standard silica-based C-band erbium-doped fiber. The tuning range covers 1451.9-1548.1 nm, with the best temperature tuning efficiency as high as 57.3 nm/ degrees C, by discretely introducing tunable fundamental-mode cutoff tapered fiber filters along a 16-m long erbium-doped fiber under a 980 nm pump power around 200 mW. The signal-ASE ratio can be higher than 45 dB whereas the FWHM of the laser lasing lights can be reduced below 0.2 nm by using an additional Fabry-Perot filter. PMID- 19550936 TI - Dramatic performance enhancement of evanescent-wave multimode fiber fluorometer using non-Lambertian light diffuser. AB - To enhance the performance of an evanescent-wave (EW) based sensor, efforts are usually made to modify the sensor architecture rather than the excitation source. In this paper, we theoretically examine the role of meridian and skew rays under total internal reflection (TIR) as well as tunneling rays with the emphasis on sensor performance. Our further investigation indicates that the intensity profile of the light source enormously influences the EW power, and thus the collectable fluorescent emission level as well. A non-Lambertian fiber-optic side emitting diffuser (FOSED) is proposed and experimentally verified, revealing that a proper alignment of this FOSED can dramatically improve the signal quality and reduce the level of stray excitation light. In particular, the adoption of a FOSED or other light diffusers with similar output profiles will ensure that the excitation power is used more efficiently, suggesting a lower demand on the excitation source power level, and the performance of the stray light filter and detector. The superiority of this innovation is further addressed by comparing it with a long period grating (LPG) fiber-optic sensor, which claims highly efficient core to cladding mode coupling. This study presents a new concept for the construction of a high-performance and cost-effective EW-based sensor system. PMID- 19550937 TI - Radiation pressure driven mechanical oscillation in deformed silica microspheres via free-space evanescent excitation. AB - Opto-mechanical effects including radiation pressure driven mechanical oscillations are observed through direct free-space evanescent excitation of whispering gallery modes in deformed silica microspheres. The simplicity, the robustness, and the relatively high coupling efficiency of free-space evanescent excitation open up new avenues for the exploration and application of opto mechanical effects. PMID- 19550938 TI - Fluorescence pH probe based on microstructured polymer optical fiber. AB - A kind of optical pH sensor was demonstrated that is based on a pH-sensitive fluorescence dye-doped (eosin) cellulose acetate (CA) thin-film modified microstructured polymer optical fiber (MPOF). It was obtained by directly inhaling an eosin-CA-acetic acid mixed solution into array holes in a MPOF and then removing the solvent (acetic acid). The sensing film showed different fluorescence intensities to different pH solutions in a pH range of 2.5-4.5. Furthermore, the pH response range could be tailored through doping a surfactant, hexadecyl trimethyl ammonium bromide (CTAB), in the sensing film. PMID- 19550939 TI - Design and modeling of tapered waveguide for photonic crystal slab coupling by using time-domain Hertzian potentials formulation. AB - This work introduces a new simulation approach to the evaluation of the time domain electromagnetic (EM) field useful in the modeling of tapered waveguide for the Photonic Crystal Slab (PCS) coupling. Only solutions of two scalar Helmholtz equations are used in the evaluation of electric and magnetic Hertzian-potentials that yields the EM field and the frequency response of the tapered waveguide. By considering simultaneously an analytical and a numerical approximation it is possible to reduce the computational burden. In order to compare the computational time we analyze the 2D structure by also using the Finite Difference Time Domain (FDTD) method and by the 3D Finite Element Method (FEM). The method is applied by starting from design criteria of the tapered structures in order to set the correct geometrical and physical parameters, and considers the field-perturbation effect in proximity of the dielectric discontinuities by generators modeling. PMID- 19550940 TI - A high speed 2x2 electro-optic switch using a polarization modulator. AB - A high speed 2x2 electro-optic switch using a polarization modulator (PolM) is proposed and experimentally demonstrated. In the proposed switch, two linearly polarized input lightwaves with orthogonal polarization directions are sent to the PolM which is connected to a polarization beam splitter (PBS). When a switching signal is applied to the PolM, the polarization directions of the two lightwaves at the output of PolM will exchange. Consequently, the lightwaves at the two output ports of the PBS would be switched, a 2x2 switch is thus realized. An optical switch with a crosstalk lower than -35 dB and a switching time less than 25 ps is experimentally demonstrated. The performance of the switch is also experimentally investigated by studying the bit error rates and eye diagrams. A technique to achieve polarization independent operation is also proposed and discussed. PMID- 19550941 TI - Numerical study for selective excitation of Ince-Gaussian modes in end-pumped solid-state lasers. AB - This study report a first systematic approach to the selective excitations of all Ince-Gaussian modes (IGMs) in end-pumped solid-state lasers. The proposed Ince Gaussian mode excitation mechanism is based on the "mode-gain control" concept. This study classifies IGMs into three categories, explores and verifies approach to excite each IGM category using numerical simulation. PMID- 19550942 TI - Stabilization of a semiconductor disk laser using an intra-cavity high reflectivity grating. AB - We demonstrate the use of a High Reflectivity Grating (HRG) as an intra-cavity element in a Semiconductor Disk Laser (or Vertical External Cavity Surface Emitting Laser) to stabilise its emission wavelength and polarization characteristics. Operation at 1058nm with up to 645mW of pump-limited output power and an M(2)~1.4 is achieved. We also show that this scheme permits tunable single-frequency operation. PMID- 19550943 TI - Inhibition of multipolar plasmon excitation in periodic chains of gold nanoblocks. AB - Periodically corrugated chains of gold nanoblocks, fabricated with high precision by electron-beam lithography and lift-off techniques, were found to exhibit optical signatures of particle plasmon states in which relative contribution of longitudinal multipolar plasmons is significantly lower than that in equivalent rectangular gold nanorods. Plasmonic response of periodic chains is dominated by dipolar plasmon modes, which in the absence of multipolar exciations are seen as background-free and spectrally well-resolved extinction peaks at infrared (IR) wavelengths. This observation may help improve spectral parameters of IR plasmonic sub-wavelength antennae. Comparative studies of plasmon damping and dephasing in corrugated chains of nanoblocks and smooth rectangular nanorods are also presented. PMID- 19550944 TI - Sensitive multiplex spectroscopy in the molecular fingerprint 2.4 mum region with a Cr(2+):ZnSe femtosecond laser. AB - An ultrashort-pulse Cr(2+):ZnSe laser is a novel broadband source for sensitive high resolution molecular spectroscopy. A 130-fs pulse allows covering of up to 380 cm(-1) spectral domain around 2.4 mum which is analyzed simultaneously with a 0.12 cm(-1) (3.6 GHz) resolution by a Fouriertransform spectrometer. Recorded in 13 s, from 70-cm length absorption around 4150 cm(-1), acetylene and ammonia spectra exhibit a 3800 signal-tonoise ratio and a 2.4.10(-7) cm(-1).Hz(-1/2) noise equivalent absorption coefficient at one second averaging per spectral element, suggesting a 0.2 ppbv detection level for HF molecule. With the widely practiced classical tungsten lamp source instead of the laser, identical spectra would have taken more than one hour. PMID- 19550945 TI - Distortion of 3D SU8 photonic structures fabricated by four-beam holographic lithography withumbrella configuration. AB - We present a quantitative study of the distortion from a threeterm diamond-like structure fabricated in SU8 polymer by four-beam holographic lithography. In the study of the refraction effect, theory suggests that the lattice in SU8 should be elongated in the [111] direction but have no distortion in the (111) plane, and each triangular-like hole array in the (111) plane would rotate by ~30 degrees away from that in air. Our experiments agree with the prediction on the periodicity in the (111) plane and the rotation due to refraction effect, however, we find that the film shrinkage during lithographic process has nearly compensated the predicted elongation in the [111] direction. In study of photonic bandgap (PBG) properties of silicon photonic crystals templated by the SU8 structure, we find that the distortion has decreased quality of PBG. PMID- 19550946 TI - Discrimination of granulocyte subtypes from light scattering: theoretical analysis using a granulated sphere model. AB - We perform extensive simulations of light scattering by a granulated sphere in the size and refractive index range of human granulated leucocytes using the discrete dipole approximation. We calculate total and depolarized side scattering signals as a function of the size and refractive indices of cell and granules, and the granule volume fraction. Using typical parameters derived from the literature data on granulocyte morphology, we show that differences between experimentally measured signals of two granulocyte subtypes can be explained solely by the difference in their granule sizes. Moreover, the calculated depolarization ratio quantitatively agrees with experimental results. We also use the Rayleigh-Debye-Gans approximation and its second order extension to derive analytical expressions for side scattering signals. These expressions qualitatively describe the scaling of signals with varying model parameters obtained by rigorous simulations, and even lead to quantitative agreement in some cases. Finally, we show and discuss the dependence of extinction efficiency and asymmetry parameter on size and volume fraction of granules. PMID- 19550947 TI - Investigation of signal dependence on tissue thickness in near infrared spectral imaging. AB - The signal intensity in near infrared autofluorescence and polarization sensitive light scattering imaging is explored as a function of tissue thickness using homogeneous porcine cardiac tissue samples as a model system. Eight images are recorded from each tissue sample including two autofluorescence images obtained under 408 nm and 633 nm excitation and six light scattering images acquired with alternating linear polarization orientations (parallel or perpendicular) under 700 nm, 850 nm, and 1000 nm linearly polarized illumination. The mean image intensity of each sample for each imaging method is plotted as a function of tissue thickness. The experimental results indicate a strong dependence of the detected signal on tissue thickness up to approximately 2 mm. Furthermore, the intensity of the spectral ratio images also exhibit thickness-dependent changes up to about 3 mm. The behavior of the light scattering experimental data was reproduced using a mathematical model based on a modified version of the random walk theory of photon migration. PMID- 19550948 TI - Dispersion characteristics of channel plasmon polariton waveguides with step trench-type grooves. AB - We have studied the dispersion characteristics of single-mode channel plasmon polaritons (CPPs) with step-trench-type groove waveguides. From the numerical simulations using the finite-element method, the modal shapes and the complex propagation constants of the CPPs over a wide spectral range were obtained. It is shown that the dispersion characteristics of the step-trench-type CPP waveguide, which is composed of a step trench with a stacking nature, show an intermediate feature between the narrow and broad trenches. The results show that this configuration allows for a well-confined CPP with a moderate propagation loss at the wavelengths investigated. PMID- 19550949 TI - Nonlinear optical phenomena in silicon waveguides: modeling and applications. AB -

Focus Serial: Frontiers of Nonlinear Optics

Several kinds of nonlinear optical effects have been observed in recent years using silicon waveguides, and their device applications are attracting considerable attention. In this review, we provide a unified theoretical platform that not only can be used for understanding the underlying physics but should also provide guidance toward new and useful applications. We begin with a description of the third-order nonlinearity of silicon and consider the tensorial nature of both the electronic and Raman contributions. The generation of free carriers through two-photon absorption and their impact on various nonlinear phenomena is included fully within the theory presented here. We derive a general propagation equation in the frequency domain and show how it leads to a generalized nonlinear Schrodinger equation when it is converted to the time domain. We use this equation to study propagation of ultrashort optical pulses in the presence of self-phase modulation and show the possibility of soliton formation and supercontinuum generation. The nonlinear phenomena of cross phase modulation and stimulated Raman scattering are discussed next with emphasis on the impact of free carriers on Raman amplification and lasing. We also consider the four-wave mixing process for both continuous-wave and pulsed pumping and discuss the conditions under which parametric amplification and wavelength conversion can be realized with net gain in the telecommunication band. PMID- 19550950 TI - Electrically switchable optical vortex generated by a computer-generated hologram recorded in polymer-dispersed liquid crystals. AB - A computer-generated hologram designed to generate an optical vortex was recorded in a cell filled with polymer-dispersed liquid crystal material under a collimated Ar+ laser beam operating at 514.5 nm. Owing to the photopolymerization induced phase separation between the polymer and the liquid crystal, an index modulation was formed between the polymer-rich and liquid crystal-rich regions. A good optical vortex beam with high fidelity was reconstructed using a collimated He-Ne laser beam. The diffraction efficiency is estimated to be about 13%-17%. With a suitable voltage applied, the reconstructed optical vortex beam can be switched owing to the index change between the polymer and the liquid crystal. PMID- 19550951 TI - Dependence of the magnetic-resonance frequency on the cut-wire width of cut-wire pair medium. AB - We investigated some magnetic metamaterials based on cut-wire pairs instead of split-ring resonators to quest for the possibility of a negative magnetic permeability. Several periodic structures of cut-wire pairs were designed and fabricated, and their transmission spectra were measured in the microwave frequency regime. The width dependence of magnetic-resonance frequency was studied both theoretically and experimentally for the periodic structures of cut wire pairs. It was found that, besides the length dependence, the magnetic resonance frequency also depends significantly on the width of cut-wire pair. A simple equivalent-circuit model proposed by Zhou et al. [Opt. Lett. 31, 3620-3622 (2006)] was employed to elucidate this interesting phenomenon and to simulate the width dependence. In the simulation the magnetic and the electric energies of the cut-wire pair were directly calculated to obtain the magnetic-resonance frequency. The theoretical results are in good agreement with the experimental ones. It reveals that there is a rather easy way to manipulate the magnetic resonance frequency of magnetic-magnetic metamaterials. PMID- 19550952 TI - Ex vivo Characterization of Atherosclerosis using Intravascular Photoacoustic Imaging. AB - The imaging of plaque composition represents one of the important steps in the interventional management of atherosclerosis. Intravascular photoacoustic (IVPA) imaging has the potential to play a major role in the detection and differentiation of an atherosclerotic lesion. The difference in the optical properties of the arterial wall and plaque constituents could be utilized to obtain high resolution photoacoustic images. In this work, through ex vivo imaging studies using a rabbit model of atherosclerosis, we evaluate the ability of IVPA imaging to detect and characterize the plaque. Specifically, the difference in the magnitude of the photoacoustic signals from the free lipids, macrophage foam cells, blood and the rest of the arterial wall were helpful in providing the contrast and detecting the fibro-cellular inflammatory plaque. The constituents identified in the IVPA images were confirmed by the results from histology. PMID- 19550953 TI - Focusing and directing of surface plasmon polaritons by curved chains of nanoparticles. AB - Excitation, focusing and directing of surface plasmon polaritons (SPPs) with curved chains of nanoparticles located on a metal surface is investigated both experimentally and theoretically. We demonstrate that, by using a relatively narrow laser beam (at normal incidence) interacting only with a portion of a curved chain of nanoparticles, one can excite an SPP beam whose divergence and propagation direction are dictated by the incident light spot size and its position along the chain. It is also found that the SPP focusing regime is strongly influenced by the chain inter-particle distance. Extensive numerical simulations of the configuration investigated experimentally are carried out for a wide set of system parameters by making use of the Green's tensor formalism and dipole approximation. Comparison of numerical results with experimental data shows good agreement with respect to the observed features in SPP focusing and directing, providing the guidelines for a proper choice of the system parameters. PMID- 19550954 TI - Facile collection of two-dimensional electronic spectra using femtosecond pulse shaping Technology. AB - This letter reports a straightforward means of collecting two-dimensional electronic (2D-E) spectra using optical tools common to many research groups involved in ultrafast spectroscopy and quantum control. In our method a femtosecond pulse shaper is used to generate a pair of phase stable collinear laser pulses which are then incident on a gas or liquid sample. The pulse pair is followed by an ultrashort probe pulse that is spectrally resolved. The delay between the collinear pulses is incremented using phase and amplitude shaping and a 2D-E spectrum is generated following Fourier transformation. The partially collinear beam geometry results in perfectly phased absorptive spectra without phase twist. Our approach is much simpler to implement than standard non collinear beam geometries, which are challenging to phase stabilize and require complicated calibrations. Using pulse shaping, many new experiments are now also possible in both 2D-E spectroscopy and coherent control. PMID- 19550956 TI - Optical ridge waveguides in SBN crystal produced by low-dose carbon ion implantation followed by a sputter etching technique. AB - This paper demonstrates, for the first time, a method to fabricate optical ridge waveguides in SBN photorefractive crystal, i.e. by first using high-energy carbon ion implantation (forming planar waveguide substrate) followed by Ar+ ion sputter etching (constructing ridged stripes). A two-dimensional (2D) cross-sectional refractive index profile of ridge waveguide is reconstructed by carefully considering the ridged topography as well as the index distributions of the planar waveguide. Based on this profile, the waveguide modes are calculated, in which shows a reasonable agreement with the experimentally observed modal near field intensity distributions. PMID- 19550955 TI - Impact of nonlinear spectral broadening in ultra-long Raman fibre lasers. AB - We present an experimental study of the impact of FWM-induced nonlinear spectral broadening on the effective reflectivity of ultra-long Raman fiber laser cavities of diverse lengths and fiber bases. We observe an exponential decay of the effective reflectivity with growing power. In standard single-mode fiber, effective reflectivity drops of up to 50% for shorter cavity lengths are observed, while the longest cavity length of 82.4km displays power leakage amounting to an effective reduction of reflectivity of approximately 30%. Using different types of fiber we examine the effect of chromatic dispersion on the Stokes wave broadening. PMID- 19550957 TI - Diffraction characteristics of a liquid crystal polarization grating analyzed using the finite-difference time-domain method. AB - This work studies the polarization characteristics of diffracted beams from a liquid crystal polarization grating. The grating is fabricated by exploiting the photo-alignment effect on a substrate that is coated with an azo dye-doped polyvinyl alcohol (PVA) film. The mechanism is induced by the irradiation of this film with suitably polarized light, which reorients the dyes. The reoriented dyes then align the liquid crystals (LCs). An LC polarization grating is fabricated using this approach. The LC alignment of the grating on one substrate is uni directionally parallel to the surface, while that on the other is rotated. The polarization and the intensity of the diffracted beams are measured. A simulation based on the finite-difference time-domain (FDTD) method is performed and is very consistent with the experimental results. PMID- 19550958 TI - Compact 90 degrees trench-based splitter for silicon-on-insulator rib waveguides. AB - Compact silicon-on-insulator (SOI) rib waveguide 90 degrees splitters based on narrow, high-aspect ratio (~10:1) trenches are designed and experimentally demonstrated. The splitter area is only 11 mum x 11 mum. Splitter optical performance is investigated as a function of both trench width and refractive index of the trench fill material. We examine three trench fill materials, air (n=1.0), SU8 (n=1.57), and index matching fluid (n=1.733), and find good agreement between experimental measurement and three dimensional (3D) finite difference time domain (FDTD) simulation. A splitting ratio of 49/51 (reflection/transmission) is measured for an index fluid-filled trench 82nm wide. PMID- 19550959 TI - Airy-Gauss beams and their transformation by paraxial optical systems. AB - We introduce the generalized Airy-Gauss (AiG) beams and analyze their propagation through optical systems described by ABCD matrices with complex elements in general. The transverse mathematical structure of the AiG beams is form-invariant under paraxial transformations. The conditions for square integrability of the beams are studied in detail. The model of the AiG beam describes in a more realistic way the propagation of the Airy wave packets because AiG beams carry finite power, retain the nondiffracting propagation properties within a finite propagation distance, and can be realized experimentally to a very good approximation. PMID- 19550960 TI - High repetition rate fiber amplifier pumped sub-20 fs optical parametric amplifier. AB - We report on a high repetition rate noncollinear optical parametric amplifier system (NOPA) based on a cavity dumped Ti:Sapphire oscillator providing the signal, and an Ytterbium-doped fiber amplifier pumping the device. Temporally synchronized NOPA pump pulses are created via soliton generation in a highly nonlinear photonic crystal fiber. This soliton is fiber amplified to high pulse energies at high repetition rates. The broadband Ti:Sapphire laser pulses are parametrically amplified either directly or after additional spectral broadening. The approach of fiber-based pump-pulse generation from a femtosecond laser, that emits in the spectral region of NOPA-gain, offers enhanced long-term stability and pulse quality compared to conventional techniques, such as signal pulse generation from a high power laser system via filamentation in bulk media. The presented system produces high-energy ultra-short pulses with pulse-durations down to 15.6 fs and pulse-energies up to 500 nJ at a repetition rate as high as 2 MHz. PMID- 19550961 TI - Frequency upconversion of multiple RF signals using optical carrier suppression for radio over fiber downlinks. AB - We propose and analyze a technique of an optical carrier transmitting two RF signals using optical carrier suppression. A single optical Mach-Zehnder modulator is used for both optical carrier suppression and signal modulation, and optical carrier suppression modulation is also used for frequency conversion of RF signals. This work shows that in contrary to the case of an optical carrier transmitting a single RF signal with optical carrier suppression where stronger optical carrier suppression improves the upconverted RF signal, weaker optical carrier suppression is preferred for an optical carrier transmitting two RF signals due to nonlinear distortion because the nonlinear distortion is reduced by using weaker optical carrier suppression. We find that the usable range of optical carrier suppression ratio is from 10 to 18 dB for RF signal upconverted to 20 GHz and beyond, and the best optical carrier suppression ratio is around 10 dB. We verify the concept and analysis with experiment. In experiment, we used two RFs at 6 and 18 GHz transmitting two 750 Mb/s signals. The experiment for the first time demonstrated that an optical carrier can transmit two RF signals using optical carrier suppression and showed that upconverted RF signals are degraded by nonlinear distortion, particularly for upconverted RF signal at 12 GHz, i.e. the RF signal at the lower frequency. PMID- 19550962 TI - Periodic and quasi-periodic non-diffracting wave fields generated by superposition of multiple Bessel beams. AB - We discuss a computer generated hologram whose transmittance is defined in terms of the Jacobi-Anger identity. If the hologram is implemented with a continuous phase spatial light modulator it generates integer-order non-diffracting Bessel beams, with a common asymptotic radial frequency, at separated propagation axes. On the other hand, when the hologram is implemented with a low-resolution pixelated phase modulator, it is possible to generate multiple Bessel beams with a common propagation axis. We employ this superposition of multiple Bessel beams to generate non-diffracting periodic and quasi-periodic wave fields. PMID- 19550963 TI - Double-clad fiber amplifier for broadband tunable ytterbium-doped oxyorthosilicates lasers. AB - Broadband tunable continuous-wave laser action and double-clad fiber amplifier of Yb:GSO, Yb:GYSO and Yb:LYSO crystals have been demonstrated. Under 940 nm diode pump, the free-running Yb:GSO, Yb:GYSO and Yb:LYSO lasers output power are 3.1 W at 1091 nm, 3.0 W at 1083 nm and 1.1 W at 1058 nm, with the corresponding optical to-optical conversion efficiency of 57%, 42% and 13%, respectively. By using intra-cavity dispersion prism and a slit to restrict the spatial mode structure, continuously tunable range extends from 1002 to 1120 nm, covering some specific wavelengths as required for metrological or optical pumping applications. By using an ytterbium-doped double-clad fiber amplifier, several watts were obtained at tunable wavelengths. Particularly, the output power was 6.5 W for Yb:GSO, 12 W for Yb:GYSO, and 4 W for Yb:LYSO at 1083 nm, which matched the atomic transition wavelength useful for optical pumping of helium. PMID- 19550964 TI - A novel kind of programmable 3(n) feed-forward optical fiber true delay line based on SOA. AB - A new kind of 3(n) feed-forward programmable optical fiber true delay line was proposed. Theoretical analysis was presented on its delay performance and expandability. Experimental demonstration was given to show the implementation of such delay lines using SOAs and Farady rotation mirrors. Delay step as small as 0.5 ps with precision of about 0.03 ps was achieved. Measurement was performed to verify the feasibility and results are given. PMID- 19550965 TI - Energy-transfer efficiency in the Er/Yb-codoped waveguide ring laser under sinusoidally modulated pump. AB - The response of an Er/Yb-codoped waveguide ring laser to a sinusoidally modulated pump power is studied. Experimentally, resonance peaks are observed and their dependences on the average pump power and the modulation index are analyzed. For high modulation indexes bistable behaviour is found. Numerically, a good agreement is obtained for the resonance peak frequencies by using a straightforward approximate model and assuming a dependence on the average pump power of the macroscopic Yb ? Er energy-transfer coefficient. This dependence can be related to these mechanisms' performance for high doping and pump levels when examined in a microscopic statistical formalism. PMID- 19550966 TI - Characteristics of switching dynamics in a semiconductor-based cavity-soliton laser. AB - The switching behavior of a semiconductor cavity soliton laser is experimentally investigated, based on a vertical-cavity surface-emitting laser with frequency selective feedback. In particular, we show the effect of frequency detuning between cavity solitons and the external injection, the temporal dynamics during ignition and erasure, and characterize the necessary injection pulse width versus its power for successful switching. PMID- 19550967 TI - Symmetric reconfigurable capacity assignment in a bidirectional DWDM access network. AB - This paper presents a novel architecture for DWDM bidirectional access networks providing symmetric dynamic capacity allocation for both downlink and uplink signals. A foldback arrayed waveguide grating incorporating an optical switch enables the experimental demonstration of flexible assignment of multiservice capacity. Different analog and digital services, such as CATV, 10 GHz-tone, 155Mb/s PRBS and UMTS signals have been transmitted in order to successfully test the system performance under different scenarios of total capacity distribution from the Central Station to different Base Stations with two reconfigurable extra channels for each down and upstream direction. PMID- 19550968 TI - Heat transport in solid and air-clad fibers for high-power fiber lasers. AB - An analytical approach for the thermal design for high-power fiber lasers and fiber components is presented. The approach is based on defining a thermal resistance for each fiber layer. Thus the importance of each layer for the heat transport is made transparent and the influence of the parameters can be studied for each layer separately. Furthermore the analysis and analytic optimization of interacting effects of groups of layers is possible. The approach is applied to air-clad-fiber with results differing up to 40 % from previous works. Furthermore the heat transport from splices is analyzed and recommendations for the thermal packaging of splices and fiber integrated components are given. PMID- 19550969 TI - Self-starting passively mode-locked chirped-pulse fiber laser. AB - We present a self-starting passively mode-locked fiber laser operating in the chirped-pulse regime for the first time. A chirped fiber Bragg grating in the cavity provides positive dispersion with negligible nonlinearity. The laser generates positively-chirped pulses with a pulse duration of 22 ps at a repetition rate of 44 MHz, which are compressible down to 1.5 ps. We believe that the presented approach reveals a pulse energy scaling potential of mode-locked fiber lasers as nonlinear effects are significantly diminished compared the other known operation regimes. PMID- 19550970 TI - Heating and rapid cooling of bulk glass after photoexcitation by a focused femtosecond laser pulse. AB - To investigate the energy dissipation process after focusing a femtosecond laser pulse inside a zinc borosilicate glass, the time-dependent lens effect in the laser focal region was observed by a transient lens (TrL) method. We found that the TrL signal after 100 ns can be explained clearly by thermal diffusion. By fitting the observed signal, we obtained the phase change due to temperature increase, the initial diameter of the heated volume and the thermal diffusivity. On the basis of the results, the temperature increase and the cooling rate were estimated to be about 1800 K and 1.7X10(8) Ks(-1), respectively. We have also observed the signal change on a 100 ns scale, which can not be explained by the thermal diffusion model. This change was attributed to the relaxation of the heated material. PMID- 19550971 TI - Three dimensional tracking for volumetric spectral-domain optical coherence tomography. AB - We present a three-dimensional (3D) tracker for a clinical ophthalmic spectral domain optical coherence tomography (SD-OCT) system that combines depth-tracking with lateral tracking, providing a stabilized reference frame for 3D data recording and post acquisition analysis. The depth-tracking system is implemented through a real-time dynamic feedback mechanism to compensate for motion artifact in the axial direction. Active monitoring of the retina and adapting the reference arm of the interferometer allowed the whole thickness of the retina to be stabilized to within +/-100 mum. We achieve a relatively constant SNR from image to image by stabilizing the image of the retina with respect to the depth dependent sensitivity of SD-OCT. The depth tracking range of our system is 5.2 mm in air and the depth is adjusted every frame.nhancement in the stability of the images with the depth-tracking algorithm is demonstrated on a healthy volunteer. PMID- 19550972 TI - Terahertz photonic crystal quantum cascade lasers. AB - We combine photonic crystal and quantum cascade band engineering to create an in plane laser at terahertz frequency. We demonstrate that such photonic crystal lasers strongly improve the performances of terahertz quantum cascade material in terms of threshold current, waveguide losses, emission mode selection, tunability and maximum operation temperature. The laser operates in a slow-light regime between the M saddle point and K band-edge in reciprocal lattice. Coarse frequency control of half of a terahertz is achieved by lithographically tuning the photonic crystal period. Thanks to field assisted gain shift and cavity pulling, the single mode emission is continuously tuned over 30 GHz. PMID- 19550973 TI - Direct electron beam writing of channel waveguides in nonlinear optical organic crystals. AB - We report on optical channel waveguiding in an organic crystalline waveguide produced by direct electron beam patterning. The refractive index profile as a function of the applied electron fluence has been determined by a reflection scan method in the nonlinear optical organic crystal 4-N, N-dimethylamino-4'-N'-methyl stilbazolium tosylate (DAST). A maximal refractive index reduction of Deltan1 = 0.3 at a probing wave-length of 633 nm has been measured for an electron fluence of 2.6mC/cm(2). Furthermore, a new concept of direct channel waveguide patterning in bulk crystals is presented and waveguiding has been demonstrated in the produced structures by end-fire coupling. Mach-Zehnder modulators have been successfully realized and a first electro-optic modulation at a wavelength of lambda= 1.55mum has been demonstrated therein. PMID- 19550974 TI - Time-gated single photon counting enables separation of CARS microscopy data from multiphoton-excited tissue autofluorescence. AB - We demonstrate time-gated confocal imaging as a means to separate coherent anti Stokes Raman scattering (CARS) microscopy data from multi-photon excited endogenous fluorescence in tissue. CARS is a quasi-instantaneous process and its signal decay time is only limited by the system's instrument response function (IRF). Signals due to two-photon-excited (TPE) tissue autofluorescence with excited state lifetimes on the nanosecond scale can be identified and separated from the CARS signal by employing time-gating techniques. We demonstrate this improved contrast on the example of CARS microscopy of intact roots of plant seedlings as well as on rat arterial tissue. PMID- 19550975 TI - Electric field enhancing properties of the V-shaped optical resonant antennas. AB - The electric field enhancing properties of the V-shaped optical resonant antenna are studied by using finite-difference time-domain method. Both dipolar and quadrupolar modes can be effectively excited and strong electric field enhancement in the gap of the V-shaped antenna is found. Compared with full-wave dipole antenna, the V-shaped antenna has a greater electric field enhancement, which can be attributed to the higher radiation directivity and the smaller curvature radius of the antenna arms. The more asymmetrical structure also contributes to the efficient quadrupolar excitation. The electric field enhancement of the V-shaped antenna has different dependences on the open angle of the V-shaped antenna for the dipolar and quadrupolar excitation. We obtained stronger electric field enhancing properties by using V-shaped bow-tie antennas, especially for the quadrupolar excitation. The V-shaped antenna and the bow-tie antenna can realize strongly localized and enhanced field and thus are well suitable for the use of near-field optics applications. PMID- 19550976 TI - Theoretical and experimental limits of cavity-dumping in passively mode-locked thin-disk oscillators. AB - The dynamical properties of a mode-locked thin disk laser with cavity-dumping in the solitary regime are studied using numerical simulations along with experimental data. Limitations of this system as well as their origin are identified. The results of these investigations agree very well with recently published experimental results. Based on these findings design criteria for future systems are deducted and estimates of possible pulse energies are made. PMID- 19550977 TI - Efficient and tolerant resonant grating coupler for multimode optical interconnections. AB - More than 60% overall coupling efficiency is achieved in the demonstrator of an optical interconnect comprising an input grating coupler, a multimode slab waveguide section and an output grating coupler. The grating coupling strength is enhanced by means of a leaky mode resonance. The efficiency of the resonant grating coupler compares favourably with the performancs reported on mirror inserts. PMID- 19550978 TI - Optical waveguide in stoichiometric lithium niobate formed by 500 keV proton implantation. AB - We report on the fabrication of planar waveguide in stoichiometric lithium niobate by 500 keV proton implantation with a dose of 1x10(17) ions/cm(2). The formation of n(e) enhancement planar waveguide in the crystal was disclosed by the dark mode spectra and the subsequent endface coupling measurement. The absorption spectra show that the postannealing treatments above 400 masculineC temperature can remove the color centers induced by implantation efficiently. The propagation loss and near-field profiles of the planar waveguide were obtained with an end-face coupling system. PMID- 19550979 TI - All silicon infrared photodiodes: photo response and effects of processing temperature. AB - CMOS compatible infrared waveguide Si photodiodes are made responsive from 1100 to 1750 nm by Si(+) implantation and annealing. This article compares diodes fabricated using two annealing temperatures, 300 and 475 degrees C. 0.25-mm-long diodes annealed to 300 degrees C have a response to 1539 nm radiation of 0.1 A W (-1) at a reverse bias of 5 V and 1.2 A W(-1) at 20 V. 3-mm-long diodes processed to 475 degrees C exhibited two states, L1 and L2, with photo responses of 0.3 +/ 0.1 A W(-1) at 5 V and 0.7 +/-0.2 A W(-1) at 20 V for the L1 state and 0.5 +/-0.2 A W(-1) at 5 V and 4 to 20 A W(-1)-1 at 20 V for the L2 state. The diodes can be switched between L1 and L2. The bandwidths vary from 10 to 20 GHz. These diodes will generate electrical power from the incident radiation with efficiencies from 4 to 10 %. PMID- 19550980 TI - Single-arm three-wave interferometer for measuring dispersion of short lengths of fiber. AB - We present a simple fiber-based single-arm spectral interferometer to measure directly the second-order dispersion parameter of short lengths of fiber (< 50 cm). The standard deviation of the measured dispersion on a 39.5-cm-long SMF28(TM) fiber is 1x10(-4) ps/nm, corresponding to 1% relative error, without employing any curve fitting. Our technique measures the second-order dispersion by examining the envelope of the interference pattern produced by three reflections: two from the facets of the test fiber and one from a mirror placed away from the fiber facet at a distance that introduces the same group delay as the test fiber at the measured wavelength. The operational constraints on system parameters, such as required bandwidth, wavelength resolution, and fiber length, are discussed in detail. Experimental verification of this technique is carried out via comparison of measurements of single mode fiber (SMF28(TM)) with published data and via comparison of measurements of a dispersion compensating fiber with those taken using conventional techniques. Moreover, we used this new technique to measure the dispersion coefficient of a 45-cm-long twin-hole fiber over a 70 nm bandwidth. It is the first time dispersion measurement on this specialty fiber is reported. PMID- 19550981 TI - Changes in the statistical properties of stochastic anisotropic electromagnetic beams on propagation in the turbulent atmosphere. AB - We report analytic formulas for the elements of the e 2 X2 cross-spectral density matrix of a stochastic electromagnetic anisotropic beam propagating through the turbulent atmosphere with the help of vector integration. From these formulas the changes in the spectral density (spectrum), in the spectral degree of polarization, and in the spectral degree of coherence of such a beam on propagation are determined. As an example, these quantities are calculated for a so-called anisotropic electromagnetic Gaussian Schell-model beam propagating in the isotropic and homogeneous atmosphere. In particular, it is shown numerically that for a beam of this class, unlike for an isotropic electromagnetic Gaussian Schell-model beam, its spectral degree of polarization does not return to its value in the source plane after propagating at sufficiently large distances in the atmosphere. It is also shown that the spectral degree of coherence of such a beam tends to zero with increasing distance of propagation through the turbulent atmosphere, in agreement with results previously reported for isotropic beams. PMID- 19550982 TI - An RZ DPSK receiver design with significantly improved dispersion tolerance. AB - We show an improved DPSK receiver design which can increase useful dispersion tolerance by up to a factor of two. The increased dispersion tolerance is achieved through optimization of the optical filter at the receiver and the delay of the Mach-Zehnder interferometer. In this paper we fully explain the concept, quantify the gain and provide an explanation for the operation of the receiver. PMID- 19550984 TI - Experimental investigation of optical beam deflection based on PLZT electro-optic ceramic. AB - Based on the optical characteristics of PLZT (Lead Lanthanum Zirconate Titanate) electro-optic ceramic, two kinds of electro-optical deflectors, triangular electrode structure and optical phased array technology, are studied in detail by using transverse electro-optic effect. Theoretically, the electro-optic deflection characteristics and mechanisms of the deflectors are analyzed; experimentally, the optical characteristics of ceramic wafer, such as the phase modulation, the hysteresis and the electro-induced loss characteristics, are measured; a systematic experiment is designed to verify the theoretical results. The effect of temperature on the performance of deflector is also investigated. Comparing with both structure's deflectors, the optical phased array technology has a higher angle scanning resolution, however, it has a relative small deflection angle. OCIS codes: (230.2090) Electro-optical devices; (160.2100) Electro-optical materials; (060.5060) Phase modulation. PMID- 19550985 TI - Controlling the influence of SPM in fiber-based chirped-pulse amplification systems by using an actively shaped parabolic spectrum. AB - We report on the experimental demonstration of the control of the influence of nonlinearity in fiber-based chirped-pulse amplification (CPA) using active spectral amplitude shaping. By applying a liquid crystal spatial light modulator, the influence of the spectral profile on the recompressed pulse quality is experimentally revealed. The parabolic spectrum is experimentally determined to be very suitable for CPA-systems in which nonlinearity is present. The corresponding nonlinear phase contribution can be efficiently compensated by a conventional grating compressor. In a proof-of-principle experiment using an Yb doped fiber- CPA-system, control at a B-integral as high as 16 rad is demonstrated. The method allows significant performance improvement of fiber based chirpedpulse amplification. PMID- 19550983 TI - Simultaneous SLO/OCT imaging of the human retina with axial eye motion correction. AB - It has been shown that transversal scanning (or en-face) optical coherence tomography (TS-OCT) represents an imaging modality capable to record high isotropic resolution images of the human retina in vivo. However, axial eye motion still remains a challenging problem of this technique. In this paper we introduce a novel method to compensate for this eye motion. An auxiliary spectral domain partial coherence interferometer (SD-PCI) was integrated into an existing TS-OCT system and used to measure accurately the position of the cornea. A light source emitting at 1310nm was used in the additional interferometer which enabled a nearly loss free coupling of the two measurement beams via a dichroic mirror. The recorded corneal position was used to drive an additional voice coil translation stage in the reference arm of the TS-OCT system to correct for axial eye motion. Currently, the correction can be performed with an update rate of ~200Hz. The TS-OCT instrument is operated with a line scan rate of 4000 transversal lines per second which enables simultaneous SLO/OCT imaging at a frame rate of 40fps. 3D data of the human retina with isotropic high resolution, that was sufficient to visualize the human cone mosaic in vivo, is presented. PMID- 19550986 TI - The effect of structural disorder on guided resonances in photonic crystal slabs studied with terahertz time-domain spectroscopy. AB - We measure the normal-incidence transmission coefficient of photonic crystal slabs with hexagonal arrays of air holes in silicon. The transmission spectra exhibit sharp resonant features with Fano line shapes. They are produced due to the coupling of the leaky photonic crystal modes, called guided resonances, to the continuum of free-space modes. We investigate the effects of several types of structural disorder on the spectra of these resonances. Our results indicate that guided resonances are very tolerant to disorder in the hole diameter and to interface roughness, but very sensitive to disorder in the lattice periodicity. PMID- 19550987 TI - Efficient femtosecond high power Yb:Lu(2)O(3) thin disk laser. AB - We demonstrate the first passively mode-locked thin disk laser based on Yb:Lu(2)O(3). The laser generates 370-fs pulses with 20.5 W of average power in a diffraction-limited beam (M(2) < 1.1). The nearly transform-limited pulses have a spectral bandwidth of 3.4 nm centered near 1034 nm. With slightly longer pulses (523 fs) we obtained 24 W of average power at a pump power of 56 W, resulting in an optical-to-optical efficiency of 43%, which is higher than for any previously mode-locked thin disk laser. PMID- 19550989 TI - Tunable luminescence of CaO-Al(2)O(3)-GeO(2) glasses. AB - We report tunable luminescence in oxygen-deficient CaO-Al(2)O(3)- GeO(2) glasses. The glass samples were prepared by adding metal Al instead of corresponding oxide (Al2O3). Efficient blue and red emissions were observed when excited by 300 and 370 nm ultraviolet light, respectively. By adjusting the content of metal Al, we could control the quantities of the defects which results in tunable luminescence from blue to white to red. Furthermore the resultant oxygen-deficient glasses have shown bright white luminescence when the excitation wavelength tuned to 335nm. Our method opens a new route for the white light illumination. PMID- 19550988 TI - Improved slow-light performance of 10 Gb/s NRZ, PSBT and DPSK signals in fiber broadband SBS. AB - We have demonstrated error-free operations of slow-light via stimulated Brillouin scattering (SBS) in optical fiber for 10-Gb/s signals with different modulation formats, including non-return-to-zero (NRZ), phase-shaped binary transmission (PSBT) and differential phase-shiftkeying (DPSK). The SBS gain bandwidth is broadened by using current noise modulation of the pump laser diode. The gain shape is simply controlled by the noise density function. Super-Gaussian noise modulation of the Brillouin pump allows a flat-top and sharp-edge SBS gain spectrum, which can reduce slow-light induced distortion in case of 10-Gb/s NRZ signal. The corresponding maximal delay-time with error-free operation is 35 ps. Then we propose the PSBT format to minimize distortions resulting from SBS filtering effect and dispersion accompanied with slow light because of its high spectral efficiency and strong dispersion tolerance. The sensitivity of the 10 Gb/s PSBT signal is 5.2 dB better than the NRZ case with a same 35-ps delay. The maximal delay of 51 ps with error-free operation has been achieved. Futhermore, the DPSK format is directly demodulated through a Gaussian-shaped SBS gain, which is achieved using Gaussian-noise modulation of the Brillouin pump. The maximal error-free time delay after demodulation of a 10-Gb/s DPSK signal is as high as 81.5 ps, which is the best demonstrated result for 10-Gb/s slow-light. PMID- 19550990 TI - Improving thin-film crystalline silicon solar cell efficiencies with photonic crystals. AB - Most photovoltaic (solar) cells are made from crystalline silicon (c-Si), which has an indirect band gap. This gives rise to weak absorption of one-third of usable solar photons. Therefore, improved light trapping schemes are needed, particularly for c-Si thin film solar cells. Here, a photonic crystal-based light trapping approach is analyzed and compared to previous approaches. For a solar cell made of a 2 mum thin film of c-Si and a 6 bilayer distributed Bragg reflector (DBR) in the back, power generation can be enhanced by a relative amount of 24.0% by adding a 1D grating, 26.3% by replacing the DBR with a six period triangular photonic crystal made of air holes in silicon, 31.3% by a DBR plus 2D grating, and 26.5% by replacing it with an eight-period inverse opal photonic crystal. PMID- 19550991 TI - Microrheology of red blood cell membranes using dynamic scattering microscopy. AB - We employ a novel optical technique, dynamic scattering microscopy (DSM), to extract the frequency dependence of the viscoelastic modulus associated with the red blood cell membrane. This approach applies the principle of dynamic light scattering to micro beads attached to the red blood cell membrane in thermal fluctuation. This allows for highthroughput characterization of a large number of cells simultaneously, which represents a significant advantage over current methods. The results in terms of the effective loss and storage moduli indicate the generic behavior of a viscoelastic material, characterized by power laws with exponents between 0 and 1. PMID- 19550992 TI - Photochromic effect in LiNbO(3): Fe :Co. AB - In this paper, Co(2)O(3) was codoped in LiNbO(3): Fe crystals. It was found that Co codoping can give rise to the strong photochromic effect in LiNbO(3): Fe. Based on the UV-VIS-NIR absorption spectra, photorefractive sensitivities, and EPR results for both virgin and sensitized states of the crystal, the photochromic mechanism in this material was suggested as follows. During sensitization, electrons transfer from O(2-) to Fe(3+) directly while holes are thermally excited from O- into the valence band and then partly trapped by Co(2+), which leads to the darkening of the crystal. In bleaching process, the electrons are excited from Fe(2+) to the conduction band by green light and recombine with the holes on the Co(2+) level. Sensitizing/bleaching experiments were also carried out in LiNbO(3): Fe: Co crystals. Fast sensitization found for this material is beneficial to two-color recording. PMID- 19550993 TI - Spectral-domain measurement of phase modal birefringence in polarization maintaining fiber. AB - We report on a new and simple method for measuring the wavelength dependence of phase modal birefringence in a polarizationmaintaining fiber. The method is based on application of a lateral pointlike force on the fiber that causes strong coupling between polarization modes and utilizes their interference resolved as the channeled spectrum. The change of the phase retrieved from two recorded channeled spectra that are associated with the known displacement of coupling point is used to determine the phase modal birefringence as a function of wavelength. A windowed Fourier transform is applied to reconstruct precisely the phase change and the phase ambiguity is removed provided that we know the phase change of the spectral fringes at one specific wavelength. The measured wavelength dependence of phase modal birefringence is compared with that resulting from the group modal birefringence measurement. PMID- 19550994 TI - All-optical M-ary ASK signal demultiplexer based on a photonic analog-to-digital conversion. AB - An all-optical M-ary amplitude shift keying (ASK) signal demultiplexer is proposed and demonstrated. It allowed us to seamlessly demultiplex a high bit rate optical M-ary ASK signal into on-off keying (OOK) signals without O/E conversion. It is composed of multilevel thresholding using self-frequency shift and OOK signal generation using optical interconnection. A level identification signal is provided as a result of multilevel thresholding and it is fed to an optical interconnection circuit which can generate corresponding OOK signals. We demonstrate the quadrature ASK signal demultiplexing at 100 Gsymbol/s and its error free operation at 10 Gb/s. PMID- 19550995 TI - Modeling focusing characteristics of low Fnumber diffractive optical elements with continuous relief fabricated by laser direct writing. AB - A theoretical model is established using Rayleigh-Sommerfeld diffraction theory to describe the diffraction focusing characteristics of low F-number diffractive optical elements with continuous relief fabricated by laser direct writing, and continuous-relief diffractive optical elements with a design wavelength of 441.6nm and a F-number of F/4 are fabricated and measured to verify the validity of the diffraction focusing model. The measurements made indicate that the spot size is 1.75mum and the diffraction efficiency is 70.7% at the design wavelength, which coincide well with the theoretical results: a spot size of 1.66mum and a diffraction efficiency of 71.2%. PMID- 19550996 TI - Anamorphic beam concentrator for linear laser-diode bar. AB - An anamorphic beam concentrator for linear laser-diode (LD) bar is presented. It consists of a tapered SiO(2)-rod with skewed and curved surface. The principle and applicability of this device are numerically investigated by ZEMAX and experimentally illustrated for the specific example of the linear LD bar. Results show that a relative symmetrical output beam spot is produced at the output facet of the rod and the intensity and spatial fluctuations in the input beam are compensated at distance of 20cm from the output facet. PMID- 19550998 TI - Intracavity transverse modes controlled by a genetic algorithm based on Zernike mode coefficients. AB - A new adaptive optics (AO) system for controlling the mode profile of a diode laser-pumped Nd:YAG solid laser has been set up in our laboratory. A 19-element piezoelectric deformable mirror (DM), which is used as the rear mirror of the solid-state laser, is controlled by a genetic algorithm (GA). To improve the system convergence rate, the GA optimizes the first 10 orders of Zernike mode coefficients rather than optimize 19 voltages on the DM. The transform matrix between the 19 voltages and the first 10 orders of Zernike mode coefficients is deduced. Comparative numerical results show that the convergence speed and the correction performance of the AO system based on optimizing Zernike mode coefficients is far better than that of optimizing voltages. Moreover, experimental results showed that this AO system could change TEM(10), TEM(11), and TEM(20) transverse modes into a TEM(00) mode successfully. PMID- 19550997 TI - 502 Watt, single transverse mode, narrow linewidth, bidirectionally pumped Yb doped fiber amplifier. AB - High power operation of narrow linewidth optical fiber amplifiers is usually limited by the onset of stimulated Brillouin scattering. In this paper, we present results demonstrating over 500 Watts of power in a single mode beam from a fiber designed to suppress stimulated Brillouin scattering through a reduction in the overlap of the optical and acoustic fields. Simulations demonstrate the potential for this fiber to achieve greater than 1000 Watts of output power. PMID- 19550999 TI - Light depolarization effects during the Freedericksz transition in nematic liquid crystals. AB - This work is aimed to the photopolarimetric characterization of the disorder evolution occurring in homeotropically aligned nematic liquid crystal films during the electrically induced Freedericksz transition. The molecular director dynamics and the transversal reorientation modes are investigated by the analysis of the depolarization of the light beam emerging from the sample. Our measurements reveal unexpected depolarization effects at the transition, which we interpret in terms of director field unhomogeneity and defects creation. PMID- 19551000 TI - Optimal multiplexed sensing: bounds, conditions and a graph theory link. AB - Measuring an array of variables is central to many systems, including imagers (array of pixels), spectrometers (array of spectral bands) and lighting systems. Each of the measurements, however, is prone to noise and potential sensor saturation. It is recognized by a growing number of methods that such problems can be reduced by multiplexing the measured variables. In each measurement, multiple variables (radiation channels) are mixed (multiplexed) by a code. Then, after data acquisition, the variables are decoupled computationally in post processing. Potential benefits of the use of multiplexing include increased signal-to-noise ratio and accommodation of scene dynamic range. However, existing multiplexing schemes, including Hadamard-based codes, are inhibited by fundamental limits set by sensor saturation and Poisson distributed photon noise, which is scene dependent. There is thus a need to find optimal codes that best increase the signal to noise ratio, while accounting for these effects. Hence, this paper deals with the pursuit of such optimal measurements that avoid saturation and account for the signal dependency of noise. The paper derives lower bounds on the mean square error of demultiplexed variables. This is useful for assessing the optimality of numerically-searched multiplexing codes, thus expediting the numerical search. Furthermore, the paper states the necessary conditions for attaining the lower bounds by a general code.We show that graph theory can be harnessed for finding such ideal codes, by the use of strongly regular graphs. PMID- 19551001 TI - Monolithically integrated laser Bragg Q-switch and wavelength converter in a PPLN crystal. AB - We report a periodically poled lithium niobate (PPLN) crystal for both temperature-insensitive laser Q-switching and temperature-tuned wavelength conversion. The PPLN crystal consists of two sections, a 20.3-mum period section functioning as an electro-optic Bragg grating for Qswitching a diode-pumped Nd:YVO4 laser at 1064 nm and a 31-mum-period section functioning as an optical parametric generator for down converting the generated 1064-nm laser. When driving the PPLN Bragg grating with 170-V voltage pulses, we measured 181 muJ pulse energy at 1064 nm from the Nd:YVO4 laser pumped by 20.4 W diode power. The 181-muJ pulsed laser was further converted into mid-infrared radiation in the monolithic PPLN crystal with 35% parametric efficiency. The wavelengths were broadly tunable in the range of 1.75-1.88 mum (signal) and 2.7-2.44 mum (idler) via temperature without affecting the performance of the PPLN Bragg Qswitch. PMID- 19551002 TI - Large dynamic range photon detector with a temperature-stabilized Si-based multi pixel photon counter. AB - We present an efficient fluorescence detector in the visible region of the spectrum with a photon detection dynamic range over 10 (6) photons/s made of a temperature-stabilized Si-based multi-pixel photon counter at temperature down to 5 degrees C. We show that effective cooling of the device by means of a compact thermo-electric cooler brings several advantages, such as high gain, low dark noise rate, and thus high signal-to-noise ratio in the efficient fluorescence detection at 398.9 nm from the (1)S(0) ? (1)P(1) transition of the ytterbium atoms in an effusive atomic beam. We present also a comparison of the fluorescence detection efficiencies between the device and a side-on photo multiplier tube with known gain positioned at the symmetric location from the ytterbium atomic beam. PMID- 19551003 TI - Ultra-compact, low RF power, 10 Gb/s silicon Mach-Zehnder modulator. AB - Silicon p(+)-i-n(+) diode Mach-Zehnder electrooptic modulators having an ultra compact length of 100 to 200 mum are presented. These devices exhibit high modulation efficiency, with a V(pi)L figure of merit of 0.36 V-mm. Optical modulation at data rates up to 10 Gb/s is demonstrated with low RF power consumption of only 5 pJ/bit. PMID- 19551004 TI - Phase locking of an array of three fiber lasers by an all-fiber coupling loop. AB - Constructing a phase-locking array of fiber lasers by an all-fiber coupling loop is presented, and the loop is composed of multiple 2x2 fiber couplers which are connected with the output ends of component fiber lasers of the array. Due to the mutual injection locking among the fiber lasers by the coupling loop, they can achieve the phase-locking states and then are coherently combined in the far field. Phase locking of an array of three fiber lasers with this configuration has been demonstrated experimentally, and obvious interference patterns and stable coherent output have been observed. PMID- 19551005 TI - Generating coherent broadband continuum soft-x-ray radiation by attosecond ionization gating. AB - The current paradigm of isolated attosecond pulse production requires a few-cycle pulse as the driver for high-harmonic generation that has a cosine-like electric field stabilized with respect to the peak of the pulse envelope. Here, we present simulations and experimental evidence that the production of high-harmonic light can be restricted to one or a few cycles on the leading edge of a laser pulse by a gating mechanism that employs time-dependent ionization of the conversion medium. This scheme enables the generation of broadband and tunable attosecond pulses. Instead of fixing the carrier-envelope phase to produce a cosine driver pulse, the phase becomes a control parameter for the center frequency of the attosecond pulse. A method to assess the multiplicity of attosecond pulses in the pulse train is also presented. The results of our study suggest an avenue towards relaxing the requirement of few-cycle pulses for isolated attosecond pulse generation. PMID- 19551006 TI - Line defects and temperature effects in liquid crystal tunable planar Bragg gratings. AB - Liquid crystal tunable planar Bragg Gratings produced by Direct UV Writing are capable of wavelength tuning of over 100GHz. However, such devices exhibit non linear tuning curves with threshold points and hysteresis. We show that these effects are due to the formation of disclination structures in the liquid crystal and discuss the role of electrode defects and sample temperature on wavelength tuning. PMID- 19551007 TI - Semilinear coherent optical oscillator with frequency shifted feedback. AB - It is shown that the saw-tooth variation of the cavity length in a photorefractive semilinear coherent oscillator can suppress the instability in the frequency domain and prevent a bifurcation in the oscillation spectrum. To achieve such a suppression the frequency of the cavity length modulation should be chosen appropriately. It depends on the photorefractive crystal parameters (electrooptic properties, photoconductivity, dimensions) and on the experimental conditions (pump intensity ratio, orientation of the pump and oscillation waves with respect to the crystallographic axes, polarization of the pump waves, etc. ). It depends also strongly on a possible misalignment of the two pump waves. On the other hand, within a certain range of the experimental parameters the mirror vibration may lead to a further frequency splitting in the already existing two mode oscillation spectrum. PMID- 19551008 TI - Frequency-doubling in femtosecond laser inscribed periodically-poled potassium titanyl phosphate waveguides. AB - Frequency doubling has been achieved in femtosecond-laser-inscribed single-mode waveguides written in two periodically-poled potassium titanyl phosphate crystals. A conversion efficiency of 0.22 %W(1) was obtained for first-order quasi-phase matching at 980 nm and an efficiency of 0.02 %W(-1) for third-order quasi-phase matching at 800 nm. PMID- 19551009 TI - Flat-top surface plasmon-polariton modes guided by double-electrode structures. AB - We characterize the frequency dependence of symmetrically-coupled long-range surface plasmon-polaritons (sc-LRSPPs) excited on double-electrode slab waveguides composed of five layers of insulator(I) and metal(M) stacked in order of IMIMI. When the core insulator has a refractive-index larger than the cladding ones, there is no cut-off core-thickness(D) for sc-LRSPP modes in all frequency range likely for modes in a conventional dielectric slab waveguide. At a specific frequency of ωc which depends on the index difference of insulator layers and the thickness of metal, the sc-LRSPP modes are non-dispersive at all for change in D. Furthermore, regardless of D alteration, the modes at ω = ω(c) consistently maintain a perfect flat-top profile in the core region and identical decay tails in the cladding. The sc-LRSPP modes with these prominent characteristics may excite an active medium sandwiched in between the metal layers very uniformly, therefore it will be interesting to implement such a non-dispersive flat-top mode for nonlinear applications of SPP waveguides. PMID- 19551010 TI - Near-field photodetection with high spatial resolution by nanocrystal quantum dots. AB - We report a new demonstration of nanoscale solution-processed photodetectors by fabricating a nano-sized gap between two electrodes and drop-casting nanocrystal quantum dots (NCQDs) into the gap. We demonstrate a detection sensitivity of 62 pW with a max responsivity of 2.7 mA/W over a device with a nano-gap of 25 nm. Additionally, we characterize the dependence of signal-to-dark current ratio and responsivity on nano-gap size. Responsivity ranges from 1 - 90 mA/W for a nano gap size range of 25 nm - 1.5 nm. Our results represent the first demonstration of how near-field optical detection for sub-diffraction nanophotonic integrated circuits can be achieved in principle using NCQDs. PMID- 19551011 TI - Introduction: physics and applications of microresonators. AB - An introduction to the Focus Issue devoted Physics and Applications of Microresonators is presented. An introduction includes a description of the area of microresontors and a description of the background and structure of the Focus Issue. PMID- 19551012 TI - Cavity opto-mechanics. AB - The coupling of mechanical and optical degrees of freedom via radiation pressure has been a subject of early research in the context of gravitational wave detection. Recent experimental advances have allowed studying for the first time the modifications of mechanical dynamics provided by radiation pressure. This paper reviews the consequences of back-action of light confined in whispering gallery dielectric microcavities, and presents a unified treatment of its two manifestations: notably the parametric instability (mechanical amplification and oscillation) and radiation pressure back-action cooling. Parametric instability offers a novel "photonic clock" which is driven purely by the pressure of light. In contrast, radiation pressure cooling can surpass existing cryogenic technologies and offers cooling to phonon occupancies below unity and provides a route towards cavity Quantum Optomechanics. PMID- 19551013 TI - High-Q nanocavity with a 2-ns photon lifetime. AB - We have succeeded in fabricating a photonic crystal nanocavity with a photon lifetime of 2.1 ns, which corresponds to a quality factor of 2.5 x 10(6). This lifetime is the longest recorded thus far in photonic crystal cavities, and was brought about by improvements in the fabrication process. Comparing our experimental quality factor with the results of calculations shows that we have suppressed variations in the radii and positions of the air holes composing a nanocavity such that their standard deviations are less than 1 nm. PMID- 19551014 TI - Near-field imaging and frequency tuning of a high-Q photonic crystal membrane microcavity. AB - We discuss experimental studies of the interaction between a nanoscopic object and a photonic crystal membrane resonator of quality factor Q=55000. By controlled actuation of a glass fiber tip in the near field of the photonic crystal, we constructed a complete spatio-spectral map of the resonator mode and its coupling with the fiber tip. On the one hand, our findings demonstrate that scanning probes can profoundly influence the optical characteristics and the near field images of photonic devices. On the other hand, we show that the introduction of a nanoscopic object provides a low loss method for on-command tuning of a photonic crystal resonator frequency. Our results are in a very good agreement with the predictions of a combined numerical/analytical theory. PMID- 19551015 TI - Polarized quantum dot emission from photonic crystal nanocavities studied under moderesonant enhanced excitation. AB - We study the linear polarization of the emission from single quantum dots embedded in an "L3" defect nanocavity in a two-dimensional photonic crystal. By using narrow linewidth optical excitation in resonance with higher-order modes, we are able to achieve strong quantum dot emission intensity whilst reducing the background from quantum dots in the surrounding lattice. We find that all the dots observed emit very strongly linearly polarized light of the same orientation as the closest mode, despite the fact that these quantum dots may be spectrally detuned by several times the mode linewidth. We discuss the coupling mechanisms which may explain this behavior. PMID- 19551016 TI - Modification of visible spontaneous emission with silicon nitride photonic crystal nanocavities. AB - Photonic crystal (PC) nanocavities based on silicon nitride membranes are studied as tools for the manipulation of spontaneous emission in the wavelength range between 550 nm and 800 nm. We observe a strong modification of the fluorescence spectrum of dye molecules spin-cast on top of the PC, indicating an efficient coupling of the dye emission to the cavity modes. The cavity design is optimized with respect to the quality factor and values of nearly 1500 are achieved experimentally. Taking into account the small mode volume, which leads to a strong Purcell enhancement, these nanocavities enable the realization of efficient single photon sources in the visible region of the spectrum. Furthermore, their fabrication is fully compatible with existing CMOS technology, making an integration into more complex optoelectronic devices feasible. PMID- 19551017 TI - Reconfigurable microfiber-coupled photonic crystal resonator. AB - We propose and demonstrate reconfigurable microfiber-coupled photonic crystal (PhC) lasers. In this generic configuration, the position of a PhC resonator can be defined (and redefined) repeatedly by simply relocating a curved microfiber along the linear PhC waveguide. In the proximity of the PhC waveguide in contact with the microfiber, the cutoff frequency (effective index) of the PhC waveguide becomes smaller (larger) than that of a bare PhC waveguide. Accordingly, when a curved microfiber is in contact with the PhC waveguide, a linear PhC resonator having Gaussian-shaped potential well is formed. Experimentally we confirm the formation of the reconfigurable resonator by observing laser operation slightly below three available band edges. PMID- 19551018 TI - High-Q cavities in multilayer photonic crystal slabs. AB - We propose a novel concept for creating high-Q cavities in photonic crystal slabs (PCS). These cavities are formed by depositing a polymer layer on top of a photonic crystal membrane fabricated in a high index semiconductor slab. We show that such multilayer structures exhibit a mode-gap and can yield high-Q microcavities with quality factors of Q approximately 106. This allows the cavity to be created by polymer processing, following the much more demanding semiconductor processing that is used to generate a uniform PCS. Depending on the polymer used, these structures can be additionally tuned using photosensitivity or the electro-optic effect. PMID- 19551019 TI - Ultra-high-Q three-dimensional photonic crystal nano-resonators. AB - Two nano-resonator modes are designed in a woodpile three-dimensional photonic crystal by the modulation of unit cell size along a low-loss optical waveguide. One is a dipole mode with 2.88 cubic half-wavelengths mode volume. The other is a quadrupole mode with 8.3 cubic half-wavelengths mode volume. Light is three dimensionally confined by a complete photonic band gap so that, in the analyzed range, the quality factor exponentially increases as the increase in the number of unit cells used for confinement of light. PMID- 19551020 TI - Optical modulation using anti-crossing between paired amplitude and phase resonators. AB - An optical modulator design based upon anti-crossing between coupled silicon microrings with independent amplitude and phase functionality is presented. The device exhibits over 25x improvement in sensitivity to an input drive signal when compared with previously studied microring modulators based on control of waveguide-resonator coupling. The new design also demonstrates an ON-OFF contrast of 14 dB, and has an ultra-compact footprint of 0.003 mm(2). The observed sensitivity enhancement suggests that this modulator may be driven directly by digital CMOS electrical signals with less than 1 V amplitude. PMID- 19551021 TI - A reconfigurable architecture for continuously variable optical slow-wave delay lines. AB - A novel reconfigurable architecture based on slow-wave propagation in integrated optical ring resonators is proposed for the realization of variable optical delay lines. A continuously variable delay is achieved by combining a coarse discrete (digital) delay, provided by a coupled resonator slow-wave structure, with a fine continuous (analog) delay given by a cascaded ring- resonator phase-shifter. The reflective configuration of the structure enables a simple, accurate and robust tuning of the delay and provides a footprint reduction by a factor 2 with respect to conventional coupled resonator optical waveguides. Proof-of-concept devices realized in 4.4% silicon oxynitride waveguides and activated by a thermal control are discussed. Experimental results demonstrate, in both spectral and time domain, a continuously variable delay, from zero to 800 ps (2 bit fractional delay), on a 2.5 Gbit/s NRZ signal, with less than 8 dB insertion loss and less than 5 mm2 device footprint. PMID- 19551022 TI - Polysilicon photonic resonators for large-scale 3D integration of optical networks. AB - We demonstrate optical microresonators in polycrystalline silicon with quality factors of 20,000. We also demonstrate polycrystalline resonators vertically coupled to crystalline silicon waveguides. Electrically active photonic structures fabricated in deposited polysilicon layers would enable the large scale integration of photonics with current CMOS microelectronics. PMID- 19551023 TI - High Q whispering gallery modes in GaAs/AlAs pillar microcavities. AB - We report the observation of whispering gallery modes (WGM) in high quality GaAs/AlAs pillar microcavities defined by electron-beam lithography and electron cyclotron resonance reactive ion etching. Photoluminescence experiments, conducted using InAs quantum dots as an internal probe, reveal a remarkably simple WGM spectrum, consisting of a single series of TE modes. For diameters ranging from 3 to 4 mum, Q-factors in excess of 15 000 were measured, allowing for WGM lasing. Noticeably, sub-micron diameter micropillars also display high Qs (~ 1000), close to the limit set by intrinsic radiative losses. These results open the way to the development of original microlasers and improved quantum-dot single photon sources. PMID- 19551024 TI - Improvement of thermal properties of ultra-high Q silicon microdisk resonators. AB - We present a detailed study of the thermal properties of ultra-high quality factor (Q) microdisk resonators on silicon-on-insulator (SOI) platforms. We show that by preserving the buried oxide layer underneath the Si resonator and by adding a thin Si pedestal layer at the interface between the resonator and the oxide layer we can increase the overall thermal conductivity of the structure while the ultra-high Q property is preserved. This allows higher field intensities inside the resonator which are crucial for nonlinear optics applications. PMID- 19551025 TI - Coupled spiral-shaped microdisk resonators with non-evanescent asymmetric inter cavity coupling. AB - We study coupled spiral-shaped microdisk resonators with non-evanescent asymmetric inter-cavity coupling via seamlessly jointed notches. Our finite difference time-domain numerical simulations reveal that the throughput-port transmissions are reciprocal between counterclockwise (CCW) and clockwise (CW) traveling-wave modes, while the drop-port transmissions and modal field distributions are input-port dependent. By introducing a slight mismatch in radii between two coupled microdisks while preserving their seamlessly jointed notches, we are able to show selectively enhanced extinction ratio for one of the split modes while suppressing the other. Our experiments using coupled spiral-shaped microdisk resonators in silicon nitride-on-silica suggest split resonances with an extinction ratio of ~20 dB using identical coupled microdisks, and an enhanced resonance extinction ratio of ~24 dB using slightly mismatched coupled microdisks. The non-evanescent coupling preserves high-Q resonances. PMID- 19551026 TI - Modal properties and modal control in vertically emitting annular Bragg lasers. AB - The modal properties, including the resonant vertical radiation, of a type of laser structures based on the annular Bragg resonance (ABR) are studied in detail. The modal threshold gains and the resonance frequencies of such lasers are obtained from the derived governing characteristic equation. Two kinds of ABR lasers, one with a pi/2 phase shift in the outer grating and the other without, are analyzed. It is numerically demonstrated that, it's possible to get a large area, high-efficiency, single defect mode lasing in ABR lasers if we choose the kind without a pi/2 phase shift in the outer grating and also a device size smaller than a critical value. PMID- 19551028 TI - Nanojets and directional emission in symmetric photonic molecules. AB - Symmetric directional emission of light from multisphere photonic molecules is experimentally shown in this work. The photonic molecules are illuminated in the vertical direction with a defocused laser beam. The emission is attributed to photonic nanojets generated in the structure. Furthermore, spectral analysis exhibit whispering gallery mode resonances of coupled and uncoupled modes. A benzene molecule-like structure consisting of a 7-microspheres cyclic photonic molecule shows a field emission pattern similar to the spatial distribution of the orbitals of the benzene molecule. PMID- 19551027 TI - Subdiffraction optical resolution of a gold nanosphere located within the nanojet of a Mie-resonant dielectric microsphere. AB - We theoretically investigate light scattering from a bi-sphere system consisting of a gold nanosphere and a lossless dielectric microsphere illuminated at a resonant optical wavelength of the microsphere. Using generalized multisphere Mie theory, we find that a gold nanosphere 100 times smaller than the dielectric microsphere can be detected with a subdiffraction resolution as fine as one-third wavelength in the background medium when the microsphere is illuminated at a Mie resonance. Otherwise, off-resonance, the spatial resolution reverts to that of the nonresonant nanojet, approximately one-half wavelength in the background medium. An important potential biophotonics application is the detection of antibody-conjugated gold nanoparticles attached to the membranes of living cells in an aqueous environment. PMID- 19551030 TI - Band formation in coupled-resonator slow-wave structures. AB - Sequences of coupled-resonator optical waveguides (CROWs) have been examined as slow-wave structures. The formation of photonic bands in finite systems is studied in the frame of a coupled oscillator model. Several types of resonator size tuning in the system are evaluated in a systematical manner. We show that aperiodicities in sequences of coupled microspheres provide an additional degree of freedom for the design of photonic bands. PMID- 19551029 TI - Percolation of light through whispering gallery modes in 3D lattices of coupled microspheres. AB - Using techniques of flow-assisted self-assembly we synthesized three-dimensional (3D) lattices of dye-doped fluorescent (FL) 5 mum polystyrene spheres with 3% size dispersion with well controlled thickness from one monolayer up to 43 monolayers. In FL transmission spectra of such lattices we observed signatures of coupling between multiple spheres with nearly resonant whispering gallery modes (WGMs). These include (i) splitting of the WGM-related peaks with the magnitude 4.0-5.3 nm at the average wavelength 535 nm, (ii) pump dependence of FL transmission showing that the splitting is seen only above the threshold for lasing WGMs, and (iii) anomalously high transmission at the WGM peak wavelengths compared to the background for samples with thickness around 25 mum. We propose a qualitative interpretation of the observed WGM transport based on an analogy with percolation theory where the sites of the lattice (spheres) are connected with optical "bonds" which are present with probability depending on the spheres' size dispersion. We predict that the WGM percolation threshold should be achievable in close packed 3D lattices formed by cavities with ~10(3) quality factors of WGMs and with ~1% size dispersion. Such systems can be used for developing next generation of resonant sensors and arrayed-resonator light emitting devices. PMID- 19551031 TI - Spectral engineering of bends and branches in microdisk coupled-resonator optical waveguides. AB - Rigorous simulations of bent and branched sections of coupled resonator optical waveguides (CROWs) composed of side-coupled whispering gallery (WG) mode microdisks are performed. Pre- and post-fabrication tuning capability of the designed structures is explored, and a novel concept of realization of tunable CROW-based routers and switches is introduced. The proposed tuning mechanism exploits the properties of CROW optical modes coupling with avoided crossing scenario rather than the previously used Vernier effect. Applications of spectrally-engineered branched CROW structures for controllable manipulation of coupling between spatially separated nano-emitters are also discussed. PMID- 19551032 TI - Guiding optical modes in chains of dielectric particles. AB - We have investigated low frequency guiding polariton modes in finite linear chains of closely packed dielectric spherical particles of different optical materials. These guiding (chain bound) modes cannot decay radiatively, because photon emission cannot take place with simultaneous conservation of energy and momentum. For extending previous work on infinite chains of spherical particles [1] and infinite rods [2, 3], we were able to apply the multisphere Mie scattering formalism to finite chains of dielectric particles to calculate quality factors of most bound modes originating from the first two Mie resonances depending on the number of particles N and the material's refractive index nr. We found that, in agreement with the earlier work [4], guiding modes exist for n(r) > 2 and the quality factor of the most bound mode scales by N(3). We interpreted this behavior as the property of "frozen" modes near the edges of guiding bands with group velocity vanishing as N increases. In contrast with circular arrays, longitudinal guiding modes in particle chains possess a higher quality factor compared to the transverse ones. PMID- 19551033 TI - Light beam output from diamond-shaped total-internal reflection modes by using intracavity air gaps. AB - We propose a novel method for extracting light beams from diamond-shaped total internal reflection modes in two-dimensional microcavity laser diodes by the use of intracavity air gaps. By fabricating such a laser diode, we experimentally demonstrate that the direction and longitudinal mode spacing of the output beams are in good accordance with theoretical calculations. PMID- 19551034 TI - On fundamental quantum noises of whispering gallery mode electro-optic modulators. AB - Using the example of whispering gallery mode (WGM) electro-optic modulator (EOM) we show that the majority of phase EOMs, particularly the resonant types, introduce additional quantum noise to the modulated light. The noise power grows quadratically with the optical power and results from the unavoidable spontaneous emission process originating from the strongly nondegenerate parametric interaction. This latter process is the physical basis for modulation. PMID- 19551035 TI - High-Q microsphere biosensor - analysis for adsorption of rodlike bacteria. AB - Theory is developed for frequency shift and linewidth-broadening induced by rodlike bacteria bound to micro-optical resonators. Optical shift of whispering gallery modes (WGMs) is modeled by introducing a form factor that accounts for random horizontal orientation of cylindrical bacteria bound by their high refractive index cell walls. Linewidth-broadening is estimated from scattering losses. Analytic results are confirmed by measurement using E.Coli as model system (~10(2) bacteria/mm(2) sensitivity), establishing the WGM biosensor as sensitive technique for detection and analysis of micro-organisms. PMID- 19551036 TI - A unique resonance mode observed in a prism-coupled micro-tube resonator sensor with superior index sensitivity. AB - Silica micro-tube resonators were studied by using a prism-coupled method and act as refractive index sensing elements. Two types of refractive index sensing modes were observed experimentally by tuning the incident angles in the prism-coupled configuration. The resonance mode with sensitivity below 100nm/RIU is a result of the typical evanescent wave interaction with the fluid, while the mode with record high sensitivity of ~ 600nm/RIU is a form of non-evanescent wave with high optical field in the low index fluid region. An analysis of the field distribution of the resonance modes in the micro-tube also revealed the existence of high order modes with strong optical field inside the low index liquid core, which leads to the high index sensitivity. Theoretical calculation of the sensitivity for this specific mode obtained by the Mie scattering method is consistent with the experimental result. A ray optics picture is presented to elucidate the physical nature of this special resonance mode observed in the micro-tube resonator. PMID- 19551038 TI - Cavity-enhanced laser absorption spectroscopy using microresonator whispering gallery modes. AB - Tunable diode laser absorption spectroscopy using microresonator whispering gallery modes (WGMs) is demonstrated. WGMs are excited around the circumference of a cylindrical cavity 125 mum in diameter using an adiabatically tapered fiber. The microresonator is very conveniently tuned by stretching, enabling the locking of an individual WGM to the laser. As the laser is scanned in frequency over an atmospheric trace-gas absorption line, changes in the fiber throughput are recorded. The experimental results of cavity-enhanced detection using such a microresonator are centimeter effective absorption pathlengths in a volume of only a few hundred microns cubed. The measured effective absorption pathlengths are in good agreement with theory. PMID- 19551037 TI - SERS-based detection in an optofluidic ring resonator platform. AB - The development of surface enhanced Raman scattering (SERS) detection has made Raman spectroscopy relevant for highly sensitive labon- a-chip bio/chemical sensors. Despite the tremendous benefit in specificity that a Raman-based sensor can deliver, development of a lab-on-a- chip SERS tool has been limited thus far. In this work, we utilize an optofluidic ring resonator (OFRR) platform to develop a SERS-based detection tool with integrated microfluidics. The liquid core optical ring resonator (LCORR) serves both as the microfluidic sample delivery mechanism and as a ring resonator, exciting the metal nanoclusters and target analytes as they pass through the channel. Using this OFRR approach and R6G as the analyte, we have achieved a measured detection limit of 400 pM. The measured Raman signal in this case is likely generated by only a few hundred R6G molecules, which foreshadows the development of a SERS-based lab-on-a-chip bio/chemical sensor capable of detecting a low number of target analyte molecules. PMID- 19551039 TI - Optimization of resonant optical sensors. AB - The sensitivity of the resonant optical sensors, which are based on measurement of the transmission and reflection spectra of optical resonators, is investigated. The following problem is addressed: When the losses of the resonator are known, what is the sharpest possible and the steepest possible shape of the resonant peaks that can be achieved experimentally? This optimization problem is solved for the case of a separated peak, which corresponds to a nondegenerated eigenvalue of the resonator. It is shown that the reflection spectrum possesses better sensitivity than the transmission spectrum. The model of the resonant sensor consisting of two coupled resonators is also considered. This model demonstrates that the sensitivity of transmission spectrum can be significantly increased by modification of the resonator structure. However, for the reflection spectrum, the best sensitivity is still given by a separated resonant peak. PMID- 19551040 TI - Nonlinear and adiabatic control of high-Q photonic crystal nanocavities. AB - This article overviews our recent studies of ultrahigh-Q and ultrasmall photonic crystal cavities, and their applications to nonlinear optical processing and novel adiabatic control of light. First, we show our latest achievements of ultrahigh-Q photonic-crystal nanocavities, and present extreme slow-light demonstration. Next, we show all-optical bistable switching and memory operations based on enhanced optical nonlinearity in these nanocavities with extremely low power, and discuss their applicability for realizing chip-scale all-optical logic, such as flip-flop. Finally, we introduce adiabatic tuning of high-Q nanocavities, which leads to novel wavelength conversion and another type of optical memories. PMID- 19551041 TI - Experimental measurement of the dispersion relations of the surface plasmon modes of metal nanoparticle chains. AB - The dispersion relations of the surface plasmon modes of metal nanoparticle chains are measured, and compared with theory. The theoretical model includes the effects of retardation, radiative damping and dynamic depolarization due to the finite size of the nanoparticles. The results reveal that, in addition to one longitudinal and one transverse mode, there is a third mode, which has not been previously reported. PMID- 19551042 TI - Simultaneous direct amplification and compression of picosecond pulses to 65-kW peak power without pulse break-up in erbium fiber. AB - Picosecond pulses at 1.56 micro mm wavelength are directly amplified with a diffraction limited beam quality in a core-pumped Er fiber with an 875 micro m(2) effective area. The interplay between nonlinear spectral broadening and anomalous fiber dispersion compresses the input pulse duration during amplification so that 42 nJ energy pulses with approximately 65 kW peak power are achieved without pulse break-up. PMID- 19551043 TI - Fusion of necklace-ring patterns into vortex and fundamental solitons in dissipative media. AB - We demonstrate that necklace-shaped arrays of localized spatial beams can merge into stable fundamental or vortex solitons in a generic model of laser cavities, based on the two-dimensional complex Ginzburg-Landau equation with the cubic quintic nonlinearity. The outcome of the fusion is controlled by the number of "beads" in the initial necklace, 2N, and its topological charge, M. We predict and confirm by systematic simulations that the vorticity of the emerging soliton is |N-M|. Threshold characteristics of the fusion are found and explained too. If the initial radius of the array (R(0)) is too large, it simply keeps the necklace shape (if R(0) is somewhat smaller, the necklace features a partial fusion), while, if R(0) is too small, the array disappears. PMID- 19551044 TI - An explanation of SRS beam cleanup in graded-index fibers and the absence of SRS beam cleanup in step-index fibers. AB - Beam cleanup via stimulated Raman scattering in multimode fibers is modeled by numerically considering the competition between the Stokes modes of graded-index and step-index fibers. The relative gain of each Stokes mode is calculated by considering the overlap the various pump and Stokes modes of the fibers. Mode competition in a graded-index fiber favors the LP(01) Stokes mode while mode competition in a step-index fiber does not favor the LP(01) Stokes mode. This model explains why beam cleanup has only been reported for graded-index fibers and not for step-index fibers. PMID- 19551045 TI - Efficient use of hybrid Genetic Algorithms in the gain optimization of distributed Raman amplifiers. AB - In this paper, we propose an efficient and accurate method that combines the Genetic Algorithm (GA) with the Nelder-Mead method in order to obtain the gain optimization of distributed Raman amplifiers. By using these two methods together, the advantages of both are combined: the convergence of the GA and the high accuracy of the Nelder-Mead. To enhance the convergence of the GA, several features were examined and correlated with fitting errors. It is also shown that when the right moment to switch between methods is chosen, the computation time can be reduced by a factor of two. PMID- 19551046 TI - Numerical simulation of self-compressed second-harmonic generation in type II potassium dihydrogen phosphate with a time predelay for Yb-doped solid-state lasers. AB - We report numerical results of second-harmonic generation in a type II potassium dihydrogen phosphate crystal with a time predelay for picosecond and/or femtosecond Yb-doped solid-state lasers, and clarify the dependence of the self compression in the second-harmonic laser pulse on the initial frequency chirp, fundamental duration and intensity, and phase-mismatching angle. We also show numerically the generation possibility of a self-compressed second-harmonic laser pulse near 20 fs. PMID- 19551047 TI - Optical monitoring and real time admittance loci calculation through polarization interferometer. AB - A simple, low coherence, vibration insensitive, polarization Fizeau interferometer is employed in this novel optical monitoring system proposed to extract the temporal phase change of the reflection coefficient of the growing film stacks. This system can directly detect fluctuating reflection coefficient and obtain the corresponding optical admittance of the growing film in real time. PMID- 19551048 TI - Chalcogenide glass microspheres; their production, characterization and potential. AB - Micro-resonators have attracted considerable attention as a potential geometry for photonic devices used in multiplexing, memory and switching. These all optical-resonators allow light at certain wavelengths to build up in intensity allowing nonlinear effects to be seen for much lower input power than in a bulk material. We report here on microspheres made from gallium-lanthanum-sulphide glass. Spheres have been produced with diameters from less than 1 mum up to 450 microm, and we demonstrate a first measured quality factor of 8x10(4) at 1.55 microm, for a chalcogenide sphere diameter of 100 microm. We also predict an ultimate Q of up to 4x10(10) at 3 microm. PMID- 19551050 TI - High-order dispersion in photonic crystal waveguides. AB - High-order dispersion in the slow-light regime of photonic crystal (PhC) waveguides was measured by utilizing integrated Mach-Zehnder interferometer (MZI) structures, and compared with theoretical results obtained from 3D plane-wave calculations. Highly accurate measurements of group-velocity dispersion (GVD), third-order dispersion (TOD) and fourth-order dispersion (FOD) at high group index (n(g)) values were enabled by minimizing external phase-distortions and increasing signal-to-noise ratio in the MZI. The experimental results for GVD, TOD, and FOD parameters at n(g) approximately 100 were approximately 10(2)ps(2)/mm, approximately 10(4)ps(3)/mm, and approximately 10(5)ps(3)/mm respectively. The results emphasize the importance of taking into consideration the effect of TOD and FOD on pulse broadening in the slow-light regime. PMID- 19551049 TI - Multiple rare earth emissions in a multicore tellurite fiber with a single pump wavelength. AB - A three-core tellurite glass fiber having different combinations of rare earth oxide dopants in each core has been fabricated using shaped die-extrusion. Three cores, doped with Ho(3+)-Tm(3+)-Yb(3+), Er(3+)-Ce(3+), and Tm(3+)-Yb(3+) respectively, exhibited visible upconversion (blue, green and red) and infrared emissions at 1.4 microm, 1.5 microm, 1.9 microm and 2.05 microm when pumped at a wavelength of 980 nm. The prospects for multi-band amplifiers and lasers are discussed. PMID- 19551051 TI - Methane detection at 1670-nm band using a hollow-core photonic bandgap fiber and a multiline algorithm. AB - The long interaction pathlengths provided by hollow-core photonic bandgap fibers (HC-PBFs) are especially advantageous for the detection of weakly absorbing gases such as methane (CH(4)). In this paper, we demonstrate methane sensing with a 1670-nm band HC-PBF. A multiline algorithm is used to fit the R(6) manifold (near 1645 nm) and, in this way, to measure the gas concentration. With this method, a minimum detectivity of 10 ppmv for the system configuration was estimated. PMID- 19551052 TI - Design of 7 and 19 cells core air-guiding photonic crystal fibers for low-loss, wide bandwidth and dispersion controlled operation. AB - We study the modal properties of feasible hollow-core photonic bandgap fibers (HC PBGFs) with cores formed by omitting either 7 or 19 central unit-cells. Firstly, we analyze fibers with thin core surrounds and demonstrate that even for large cores the proposed structures are optimum for broad-band transmission. We compare these optimized structures with fibers which incorporate antiresonant core surrounds which are known to have low-loss. Trade-offs between loss and useful bandwidth are presented. Finally, we study the effects that small modifications to the core surround have on the fiber's group velocity dispersion, showing the possibility of engineering the dispersion in hollow-core photonic bandgap fibers. PMID- 19551053 TI - Holographic injection locking of a broad area laser diode via a photorefractive thin-film device. AB - We demonstrate locking of a high power broad area laser diode to a single frequency using holographic feedback from a photorefractive polymer thin-film device for the first time. A four-wave mixing setup is used to generate feedback for the broad area diode at the wavelength of the single frequency source (Ti:Sapphire laser) while the spatial distribution adapts to the preferred profile of the broad area diode. The result is an injection-locked broad area diode emitting with a linewidth comparable to the Ti:Sapphire laser. PMID- 19551054 TI - Holography with extended reference by autocorrelation linear differential operation. AB - We introduce a generalization of Fourier transform holography that allows the use of the boundary waves of an extended object to act as a holographic-like reference. By applying a linear differential operator on the field autocorrelation, we use a sharp feature on the extended reference to reconstruct a complex-valued image of the object of interest in a single-step computation. We generalize the approach of Podorov et al. [Opt. Express 15, 9954 (2007)] to a much wider class of extended reference objects. Effects of apertures in Fourier domain and imperfections in the reference object are analyzed. Realistic numerical simulations show the feasibility of our approach and its robustness against noise. PMID- 19551055 TI - Propagation of a cosh-Gaussian beam through an optical system in turbulent atmosphere. AB - The propagation of a cosh-Gaussian beam through an arbitrary ABCD optical system in turbulent atmosphere has been investigated. The analytical expressions for the average intensity at any receiver plane are obtained. As an elementary example, the average intensity and its radius at the image plane of a cosh-Gaussian beam through a thin lens are studied. To show the effects of a lens on the average intensity and the intensity radius of the laser beam in turbulent atmosphere, the properties of a collimated cosh-Gaussian beam and a focused cosh-Gaussian beam for direct propagation in turbulent atmosphere are studied and numerically calculated. The average intensity profiles of a cosh-Gaussian beam through a lens can have a shape similar to that of the initial beam for a longer propagation distance than that of a collimated cosh-Gaussian beam for direct propagation. With the increment in the propagation distance, the average intensity radius at the image plane of a cosh-Gaussian beam through a thin lens will be smaller than that at the focal plane of a focused cosh-Gaussian beam for direct propagation. Meanwhile, the intensity distributions at the image plane of a cosh-Gaussian beam through a lens with different w(0) and Omega(0) are also studied. PMID- 19551056 TI - Generation of optical vortex beams by nonlinear wave mixing. AB - It is shown that optical vortex beams can be generated from a non-vortex fundamental beam by an optical frequency conversion process taking place within a twisted nonlinear photonic crystal. This is done without any first-order (linear) refractive optics. Through such a proposed structure, all-optical switching of vortices with different helicities is made possible, as well as the simultaneous application of counter-rotating vortex beams of different frequencies. PMID- 19551057 TI - Threshold for stimulated Brillouin scattering in optical fiber. AB - We show that in optical fiber the threshold exponential gain, Gth, for stimulated Brillouin scattering initiated by spontaneous Brillouin scattering is functionally and strongly dependent on the material, length and numerical aperture of the fiber and the pump wavelength. For silica fiber we show that the value of G(th) at lambda congruent with 1 microm ranges from as low as approximately 5 in long fiber ( > or =few kms) to ~10-12 in fibers of approximately 100 m length and approximately 20-23 for very short fibers (<10 cm). PMID- 19551058 TI - Chirp-enhanced fast light in semiconductor optical amplifiers. AB - We present a novel scheme to increase the THz-bandwidth fast light effect in semiconductor optical amplifiers and increase the number of advanced pulses. By introducing a linear chirp to the input pulses before the SOA and recompressing at the output with an opposite chirp, the advance-bandwidth product reached 3.5 at room temperature, 1.55 microm wavelength. This is the largest number reported, to the best of our knowledge, for a semiconductor slow/fast light device. PMID- 19551059 TI - Bend loss in structured optical fibres. AB - Bend loss characterisation tests are carried out in two air-silica structured fibres: a periodic photonic crystal fibre and a non-periodic fibre. An explanation based on resonant coupling between interstitial zones accounts for the improved confinement of non-periodic structured optical fibres. PMID- 19551060 TI - Light coupling between two parallel CO2-laser written long-period fiber gratings. AB - We demonstrate experimentally the light coupling effects between two parallel CO(2)-laser written long-period fiber gratings. For gratings written in standard single-mode fibers, the coupling efficiency depends strongly on the fiber orientation with the strongest coupling obtained when the exposed sides of the fibers face each other, while for gratings written in boron-doped fibers, the coupling efficiency is independent of the fiber orientation. We achieve a peak coupling efficiency of approximately 86% with gratings written in boron-doped fibers by using a suitable surrounding refractive index and offset distance between the two gratings. Our results suggest the possibility of realizing efficient broadband all-fiber couplers with CO(2)-laser written long-period fiber gratings. PMID- 19551061 TI - Multichannel balanced electro-optic detection for Terahertz imaging. AB - We present THz imaging with 1D electro-optic sampling of ultrashort THz pulses by multichannel balanced detection. Using a lock-in technique, it combines the advantage of a high signal to noise ratio along with the fast acquisition time of multichannel detection. The object is probed by a line focus and the resolution can be adjusted. The performance of the system is demonstrated exemplarily by imaging two objects. PMID- 19551062 TI - Optimal self-referenced sensing using long- and short- range surface plasmons. AB - Dual-mode surface-plasmon resonance (SPR) sensors use both long- and short- range surface plasmon waves to differentiate surface binding interactions from interfering bulk effects. We have optimized the design of these sensors for minimum surface limit of detection (LOD) using a Cramer-Rao lower bound for spectral shift estimation. Despite trade-offs between resonance width, minimum reflectivity, and sensitivity for the two modes, a range of reasonable design parameters provides nearly optimal performance. Experimental verification using biotin-streptavidin binding as a model system reveals that sensitivity and LOD for dual-mode sensors remains competitive with single-mode sensors while compensating for bulk effects. PMID- 19551063 TI - Observation of transverse interference fringes on an atom laser beam. AB - Using the unique detection properties offered by metastable helium atoms we have produced high resolution images of the transverse spatial profiles of an atom laser beam. We observe fringes on the beam, resulting from quantum mechanical interference between atoms that start from rest at different transverse locations within the outcoupling surface and end up at a later time with different velocities at the same transverse position. Numerical simulations in the low output-coupling limit give good quantitative agreement with our experimental data. PMID- 19551064 TI - Optical probe based on double-clad optical fiber for fluorescence spectroscopy. AB - We report an optical probe based on a single double-clad fiber (DCF), which is suitable for fluorescence spectroscopy. The excitation light is delivered through the small diameter core of the DCF and the excited fluorescence light is collected by the large diameter inner cladding of the same fiber. To retrieve the signal beam from the inner cladding, we utilize a DCF coupler that couples only the light beams in the inner claddings of two different pieces of DCF. It was found that the separation of the channel for the excited beam from the channel for the excitation beam in the same piece of fiber could diminish the autofluorescence background noise generated by the fiber itself, while maintaining all the benefits of a single-fiber probe system. The usefulness of the DCF probe and the performance of the DCF coupler are then reported by presenting the fluorescence spectrum of a fresh gingko leaf and comparing it with the spectrum taken with conventional methods. The fabrication process of the DCF fiber and the inner cladding mode coupler for it are also presented. PMID- 19551065 TI - Quantitative polarized phase microscopy for birefringence imaging. AB - A novel application of quantitative phase imaging under linearly polarized light is introduced for studying unstained anisotropic live cells. The method is first validated as a technique for mapping the two-dimensional retardation distribution of a well-characterized optical fiber and is then applied to the characterization of unstained isolated cardiac cells. The experimental retardation measurements are in very good agreement with the established Brace-Kohler method, and additionally provide spatially resolved cell birefringence and phase data. PMID- 19551066 TI - Absorption-induced trapping in an anisotropic magneto-optical trap. AB - We report on a simple anisotropic magneto-optical trap for neutral atoms that produces a large sample of cold atoms confined in a cylindrically-shaped volume with a high aspect ratio (100:1). Due to the large number of trapped atoms, the laser beams that propagate along the optically thick axis of the trap to cool the atoms are substantially attenuated. We demonstrate that the resulting intensity imbalance produces a net force that spatially localizes the atoms. This limits both the trap length and the total number of trapped atoms. Rotating the cooling beams by a small angle relative to the trap axis avoids the problem of attenuation, and atoms can be trapped throughout the entire available trapping volume. Numerical and experimental results are reported that demonstrate the effects of absorption in an anisotropic trap, and a steady-state, line-center optical path length of 55 is measured for a probe beam propagating along the length of the trap. PMID- 19551067 TI - When does the choice of the refractive index of a linear, homogeneous, isotropic, active, dielectric medium matter? AB - Two choices are possible for the refractive index of a linear, homogeneous, isotropic, active, dielectric material. Either of the choices is adequate for obtaining frequency-domain solutions for (i) scattering by slabs, spheres, and other objects of bounded extent; (ii) guided--wave propagation in homogeneously filled, cross-sectionally uniform, straight waveguide sections with perfectly conducting walls; and (iii) image formation due to flat lenses. The correct choice does matter for the half-space problem, but that problem is not realistic. PMID- 19551068 TI - Characterization of frequency noise on a broadband infrared frequency comb using optical heterodyne techniques. AB - We measure the frequency noise across a Cr:forsterite infrared frequency comb through the optical heterodyne beat of different comb teeth against stable continuous wave (CW) lasers. This sensitive measurement shows strong correlations of the frequency noise between spectral components of the comb, relative to a fixed optical frequency near the 1.3 micron carrier of the Cr:forsterite laser. The correlated frequency fluctuations are shown to arise from amplitude noise on the pump laser. We also report a preliminary comparison of excess noise that occurs during supercontinuum generation in both highly nonlinear fiber and an extruded glass microstructured fiber. PMID- 19551070 TI - Optical scattering resonances of single and coupled dimer plasmonic nanoantennas. AB - The optical resonances of individual plasmonic dimer antennas are investigated using confocal darkfield spectroscopy. Experiments on an array of antennas with varying arm lengths and interparticle gap sizes show large spectral shifts of the plasmon modes due to a combination of geometrical resonances and plasmon hybridization. The resonances of the coupled-dimer antennas are considerably broadened compared to those of single nanorods, which is attributed to a superradiant damping of the coupled antenna modes. The scattering spectra are compared with electrodynamic model calculations that demonstrate both the near field and far-field characteristics of a half-wave antenna. PMID- 19551069 TI - Dispersion, birefringence, and amplification characteristics of newly designed dispersion compensating hole-assisted fibers. AB - We propose a new design of hole-assisted fiber (HAF) that can compensate for the accumulated dispersion in single-mode fiber link along with dispersion slope, thus providing broadband dispersion compensation over C-band as well as can amplify the signal channels by utilizing the stimulated Raman scattering phenomena. The proposed dispersion-compensating HAF (DCHAF) exhibits the lowest dispersion coefficient of -550 ps/nm/km at 1550 nm with an effective mode area of 15.6 microm(2). A 2.52 km long module of DCHAF amplifies incoming signals by rendering a gain of 4.2 dB with +/-0.8 dB gain flatness over whole C-band. To obtain accurate modal properties of DCHAF, a full-vector finite element method (FEM) solver is employed. The macro-bend loss characteristics of the proposed DCHAF are evaluated using FEM solver in cylindrical coordinate systems of a curved DCHAF, and low bending losses (<10(-2) dB/m for 1 cm bending radius) are obtained for improved DCHAF design while keeping intact its dispersion compensation and Raman amplification properties. We have further investigated the birefringence characteristics that can give significant information on the polarization mode dispersion of DCHAF by assuming a certain deformation (eccentricity e = 7%) either in air-holes or in the doped core or in both at a same time. It is noticed that the distortion in air-holes induces a birefringence of 10(-5), which is larger by a factor of 10 than the birefringence caused due to the core ellipticity. A PMD of 11.3 ps/ radicalkm is obtained at 1550 nm for distorted air-holes DCHAF structure. PMID- 19551071 TI - Wideband tunable wavelength-selective coupling in asymmetric side-polished fiber coupler with dispersive interlayer. AB - We demonstrate tunable highly wavelength-selective filter based on a 2 x 2 asymmetric side-polished fiber coupler with dispersive interlayer in one of the coupling arms. The asymmetric fiber coupler is made of two side-polished fibers using identical single-mode fibers and one of the polished fibers is further chemically etched at the central evanescent coupling region to gain closer to the core. An optical liquid with different dispersion characteristics than that of silica fiber is used to fill up the etched hollow and therefore the propagation constant for the polished fiber with dispersive liquid becomes more dispersive and crosses with that of another untreated polished fiber. The location of the cross point and the cross angle between two propagation constant curves determine the coupling wavelength and coupling bandwidth as well as channel wavelength separation, respectively. The coupling wavelength can be tuned at least wider than 84 nm (1.326-1.410 microm) under index variation of 0.004 and with coupling ratios of higher than 30 dB. PMID- 19551072 TI - Physical origin of the high energy optical response of three dimensional photonic crystals. AB - The physical origin of the optical response observed in three-dimensional photonic crystals when the photon wavelength is equal or lower than the lattice parameter still remains unsatisfactorily explained and is the subject of an intense and interesting debate. Herein we demonstrate for the first time that all optical spectra features in this high energy region of photonic crystals arise from electromagnetic resonances within the ordered array, modified by the interplay between these resonances with the opening of diffraction channels, the presence of imperfections and finite size effects. All these four phenomena are taken into account in our theoretical approach to the problem, which allows us to provide a full description of the observed optical response based on fundamental phenomena as well as to attain fair fittings of experimental results. PMID- 19551073 TI - Enhanced optical nonlinearity in amorphous silicon and its application to waveguide devices. AB - We present the first measurements of optical nonlinearity due to free carrier effects in amorphous silicon films using z-scan technique, demonstrating enhanced nonlinearity due to existence of midgap localized states. We also introduce, fabricate and experimentally characterized a new composite waveguide structure consisting of amorphous and crystalline silicon. The fabricated composite rib waveguide confirms to have enhanced free-carrier nonlinearity at the estimated value of 4cm/GW, i.e., seven times larger than that of a crystalline silicon waveguide. Due to existence of the midgap localized states in amorphous silicon, the measured free-carrier lifetime in the composite rib waveguide was about approximately 300ps which is shorter than the values reported in the literature for similar geometries made of silicon. PMID- 19551074 TI - Scattering characteristics of simplified cylindrical invisibility cloaks. AB - The previously reported simplified cylindrical linear cloak is improved so that the cloak's outer surface is impedance-matched to free space. The scattering characteristics of the improved linear cloak is compared to the previous counterpart as well as the recently proposed simplified quadratic cloak derived from quadratic coordinate transformation. Significant improvement in invisibility performance is noticed for the improved linear cloak with respect to the previously proposed linear one. The improved linear cloak and the simplified quadratic cloak have comparable invisibility performances, except that the latter however has to fulfill a minimum shell thickness requirement (i.e. outer radius must be larger than twice of inner radius). When a thin cloak shell is desired, the improved linear cloak is much more superior than the other two versions of simplified cloaks. PMID- 19551075 TI - Planar glass waveguide ring resonators with gain. AB - The frequency resolution of an active waveguide ring resonator spectrometer is fundamentally limited by spontaneous emission noise produced by the gain medium. A closed-form expression for this resolution is derived, and the result is used to determine the minimum, rms, angular rotation rate, random walk error achievable by an active ring resonator gyroscope. An active waveguide ring resonator is demonstrated in a neodymium-doped glass, and a finesse of 250 at a signal wavelength of 1060 nm is achieved for the 1.6 cm diameter ring under laser diode pumping. This finesse corresponds to an effective propagation loss on the order of 0.013 dB/cm, which is the lowest value reported to date for rings of this size. PMID- 19551076 TI - Wavelength-multiplexed submicron holograms for disk-compatible data storage. AB - By using a hybrid diffractive and refractive achromat with extended depth of focus, we have successfully recorded a micro-hologram array with diffraction limited individual spot size maintained throughout the thickness of recording medium. An electrically programmable wavelength combiner was constructed in which a white light source was adopted. By modifying on a commercial CD readout head, we configured a compact micro-hologram recording/readout system that is compatible to existing disk storage technology. Base on the wavelength combiner and recording/readout system, wavelength-multiplexed micro-holograms were recorded and recovered. The presented results demonstrate the practicality of our novel storage architecture. PMID- 19551077 TI - Extreme shearing interferometry: theoretical limits with practical consequences. AB - In this work we analyze the frequency response, the spatial distribution and continuity of the recovered phase in Lateral Shearing Interferometry (LSI). This frequency content and topology of the recovered phase is analyzed for the forward LSI operator as well as its inverse LSI operator using one, two, or n two dimensional sheared interferograms. The spatial frequency response of the shearing interferometer is well known and for the reader's convenience, it is briefly revisited in a new perspective. It is however less well-known and more interesting to analyze the spatial distribution of the sheared data as well as the spatial topology of the recovered phase produced by some inverse LSI operators. Also we define a useful space of functions S with the property that any sheared data available, along any direction, may be used to recovered a smooth continuous phase with the bonus property of fully covering the pupil of the wavefront being tested. These combined aspects allow us to find the best possible wave-front reconstruction from the available sheared data using one, two or n sheared interferograms. PMID- 19551078 TI - Detection of quantum-dot labelled proteins using soft glass microstructured optical fibers. AB - The detection of quantum-dot labeled proteins is demonstrated within lead silicate soft glass microstructured optical fibers using near infrared light. The protein concentration is measured using a new fluorescence capture approach. Light guided within the fiber is used both to excite and collect fluorescent photons, and the detection limit achieved without optimization of the fiber geometry is 1 nM, using just 3% of the guided mode of the fiber. Issues that currently restrict the detection of lower protein concentrations are discussed. PMID- 19551079 TI - Three dimensional analysis of chromatic aberration in diffractive elements with extended depth of focus. AB - The paper presents the polychromatic analysis of two diffractive optical elements with extended depth of focus: the linear axicon and the light sword optical element. Chromatic aberration produces axial displacement of the focal segment line. Thus, we explore the possibility of extending the focal depth of these elements to permit superposition of the chromatic foci. In the case of an axicon, we achieve an achromatic zone where focusing is produced. In the case of the light sword element, we show that the focusing segment is out of axis. Therefore a superposition of colors is produced, but not on axis overlapping. Instead, three colored and separated foci are simultaneously obtained in a single plane. Three dimensional structures of the propagated beams are analyzed in order to provide better understanding of the properties and applications of such elements. PMID- 19551080 TI - Control of ionization processes in high band gap materials via tailored femtosecond pulses. AB - Control of two basic ionization processes in dielectrics i.e. photo ionization and electron-electron impact ionization on intrinsic time and intensity scales is investigated experimentally and theoretically. Temporally asymmetric femtosecond pulses of identical fluence, spectrum and pulse duration result in different final free electron densities. We found that an asymmetric pulse and its time reversed counterpart address two ionization processes in a different fashion. This results in the observation of different thresholds for surface material modification in sapphire and fused silica. We conclude that control of ionization processes with tailored femtosecond pulses is suitable for robust manipulation of breakdown and thus control of the initial steps of laser processing of high band gap materials. PMID- 19551081 TI - Optical transmission through single subwavelength apertures using prism coupled input of laser light of annular intensity profile. AB - Light transmission through a single subwavelength aperture in a silver film is examined with a novel input configuration comprising an annular laser beam of variable diameter that is prism-coupled to the back face of the silver. Transmission peaks driven by excitation of the back-face surface plasmon mode or by the aperture resonance itself are separately observed. For both cases, comparison of films with and without a front-face, circular, grating implies significantly more efficient coupling from the aperture fields to the front-face surface plasmon than directly to free radiation. PMID- 19551083 TI - Magnetic and electric excitations in split ring resonators. AB - We studied the electric and magnetic resonances of U-shaped SRRs. We showed that higher order excitation modes exist in both of the electric and magnetic resonances. The nodes in the current distribution were found for all the resonance modes. It turns out that the magnetic resonances are the modes with odd number of half-wavelength of the current wave, i.e. lambda/2, 3lambda/2 and 5lambda/2 modes, and the electric resonances are modes with integer number of whole-wavelength of current wave, i.e. lambda, 2lambda and 3lambda modes. We discussed the electric moment and magnetic moment of the electric and magnetic resonances, and their dependence to the length of two parallel side arms. We show that the magnetic moment of magnetic resonance vanishes as the length of side arms of the SRR reduces to zero, i.e. a rod does not give any magnetic moment or magnetic resonance. PMID- 19551082 TI - Laser emission in proton-implanted Nd:YAG channel waveguides. AB - The performance of lasers based on channel waveguides produced by proton implantation in Nd:YAG crystals through an electroformed mask is reported. The fabrication method used can produce several waveguide lasers in the crystal by a single implantation process with very good optical performance. The analysis and comparison of the main laser emission features, as well as the propagation losses of these waveguides, by using different output couplers in the laser cavity is also presented. PMID- 19551084 TI - Enhancement of fluorescence-based sensing using microstructured optical fibres. AB - We develop a generic model of excitation and fluorescence recapturing within filled microstructured optical fibres (MOFs) with arbitrary structure and demonstrate that the light-matter overlap alone does not determine the optimal fibre choice. Fibre designs with sub-wavelength features and high-index glasses exhibit localised regions of high intensity, and we show that these regions can lead to approximately two orders of magnitude enhancement of fluorescence recapturing. Here we show how this regime can be exploited for sensing and demonstrate experimentally in-fibre excitation and fluorescence recapturing within a filled, solid-core MOF. PMID- 19551085 TI - Systematic comparison of the discrete dipole approximation and the finite difference time domain method for large dielectric scatterers. AB - We compare the discrete dipole approximation (DDA) and the finite difference time domain (FDTD) method for simulating light scattering of spheres in a range of size parameters x up to 80 and refractive indices m up to 2. Using parallel implementations of both methods, we require them to reach a certain accuracy goal for scattering quantities and then compare their performance. We show that relative performance sharply depends on m. The DDA is faster for smaller m, while the FDTD for larger values of m. The break-even point lies at m = 1.4. We also compare the performance of both methods for a few particular biological cells, resulting in the same conclusions as for optically soft spheres. PMID- 19551086 TI - All-optical axially multi-regional super resolved imaging. AB - In this paper we present a new approach of all-optical extended depth of focus providing two (or more) discrete ranges of focused imaging for close as well as far ranges. The fact that the extended depth of focus is not continuous allows obtaining improved contrast in the two (or more) axial regions of extended depth of focus. The design is aimed for the cell phone camera applications where dual range extended depth of focus can allow simultaneous reading of business cards at very short distance as well as very high contrasted imaging at far range. PMID- 19551088 TI - Active polarization stabilization in optical fibers suitable for quantum key distribution. AB - Polarization feedback control of single-photon pulses has been achieved in long distance fibers for more than 10 hours, which facilitated "one-way" polarization encoded quantum key distribution with long-term stabilities. Experimental test of polarization encoding in 75 km fibers demonstrated that the single-photon polarization transformation in long-distance fibers could be controlled to provide a typical QBER of (3.9+/-1.5)% within a long-term operation of 620 minutes. PMID- 19551087 TI - Ultrafast all-optical switching with low saturation energy via intersubband transitions in GaN/AlN quantum-well waveguides. AB - A fiber-optic pump-probe setup is used to demonstrate all-optical switching based on intersubband cross-absorption modulation in GaN/AlN quantum-well waveguides, with record low values of the required control pulse energy. In particular, a signal modulation depth of 10 dB is obtained with control pulse energies as small as 38 pJ. Such low power requirements for this class of materials are mainly ascribed to an optimized design of the waveguide structure. At the same time, the intersubband absorption fully recovers from the control-pulse-induced saturation on a picosecond time scale, so that these nonlinear waveguide devices are suitable for all-optical switching at bit rates of several hundred Gb/s. PMID- 19551089 TI - Optimal design of omni-directional viewing angle switching panel. AB - A viewing angle switching (VAS) panel used for the conversion of wide and narrow viewing angle has been demonstrated. Unlike the previous VAS panels, this one features a pair of same-handed circular polarizers and it is capable of switching the viewing angle omni-directionally from wide to narrow or vice versa by electrically controlling the birefringence effect of the LC layer which is sandwiched in between. To explain how it works, the design principle as well as the optimization of both wide and narrow viewing modes will be elaborated hereinafter. PMID- 19551090 TI - Perfect 4-way splitting in nano plasmonic X-junctions. AB - A plasmon wave is perfectly split to 4 identical waves when encountering nano intersection. This is substantially different from the dielectric waveguides case where power coupling to vertical segments is negligible. When larger multimode plasmonic junction is realized--beating and retardation come into effect. The analysis of the plasmonic coupling in this device is helpful also in understanding plasmonic assisted enhanced transmission. PMID- 19551091 TI - Slow light with flat or offset band edges in few-mode fiber with two gratings. AB - We consider mode coupling in multimode optical fibers using either two Bragg gratings or a Bragg grating and a long-period grating. We show that the magnitude of the band edge curvature can be controlled leading to a flat, quartic band-edge or to two band edges at distinct, nonequivalent k-values, allowing precise control of slow light propagation. PMID- 19551092 TI - Two-photon four-qubit cluster state generation based on a polarization-entangled photon pair. AB - We propose and experimentally demonstrate a two-photon four-qubit cluster state generator using linear optics and a single polarization-entangled photon-pair source based on spontaneous parametric down-conversion (SPDC). Our novel scheme provides greater design flexibility compared to previous schemes that rely on hyperentanglement. PMID- 19551093 TI - Horizontal single and multiple slot waveguides: optical transmission at lambda = 1550 nm. AB - We experimentally demonstrate the optical transmission at 1550 nm of the fundamental slot modes (quasi-TM modes) in horizontal single and multiple slot waveguides and ring resonators consisting of deposited amorphous silicon and silicon dioxide. We demonstrate that the horizontal multiple slot configuration provides enhanced optical confinement in low index slot regions compared to a horizontal single slot structure with the same total SiO2 layer thickness by comparing their thermo-optic coefficients for the horizontal slot ring resonators. We show in these early structures that horizontal slot waveguides have low propagation loss of 6 approximately 7 dB/cm. The waveguide loss is mainly due to a-Si material absorption. The addition of a-Si/SiO(2) interfaces does not introduce significant scattering loss in a horizontal multiple slot waveguide compared to a horizontal single slot waveguide. PMID- 19551094 TI - Oval-like hollow intensity distribution of tightly focused femtosecond laser pulses in air. AB - The propagation of a tightly focused femtosecond laser pulse in air has been investigated. Unlike long-distance self-guided propagation of short laser pulses, a novel oval-like hollow distribution of the laser intensity is observed in the experiments and reproduced by the numerical simulations. The formation of the hollow structures can be explained by the interplay between ionization-induced refraction and Kerr self-focusing. PMID- 19551095 TI - Triple-wavelength erbium fiber ring laser based on compound-ring scheme. AB - .A triple-wavelength erbium-doped compound ring fiber laser using the fiber-based triple-ring filter (TRF) is proposed and experimentally investigated. Using the fiber-based TRF laser scheme, the proposed laser can lase three wavelengths simultaneously. The fiber laser retrieve the optical side-mode suppression ratios (SMSRs) of 40.2, 40.4 and 41.6 dB and the output powers of -9, -8.8 and -7.6 dBm at the wavelengths 1555.89, 1556.77 and 1557.66 nm, respectively. The mode spacing of the triple-wavelength fiber laser is nearly 0.9 nm. Moreover, the output power stability of the ring laser has also been measured and analyzed. PMID- 19551096 TI - Multiple quasi-phase-matching for enhanced generation of selected high harmonics in aperiodic modulated fibers. AB - A technique for enhanced generation of selected high harmonics in a gas medium, in a high ionization limit, is proposed in this paper. An aperiodically corrugated hollow-core fiber is employed to modulate the intensity of the fundamental laser pulse along the direction of propagation, resulting in multiple quasi-phase-matched high harmonic emissions at the cutoff region. Simulated annealing (SA) algorithm is applied for optimizing the aperiodic hollow-core fiber. Our simulation shows that the yield of selected harmonics is increased equally by up to 2 orders of magnitude compared with no modulation and this permits flexible control of the quasi-phase-matched emission of selected harmonics by appropriate corrugation. PMID- 19551097 TI - Spontaneous emission in GaN/InGaN photonic crystal nanopillars. AB - We investigate the physics of spontaneous emission in a photonic crystal (PhC) made of GaN rods with embedded InGaN quantum wells, formed on a thick GaN layer. Although the PhC lies on a higher-index medium, we evidence the existence of unexpected quasi-guided Bloch modes which are strongly localized in the PhC region and possess a long lifetime. These modes determine the behavior of spontaneous emission such as the emission diagram and Purcell effect, as would happen in the usual case of emission in a PhC membrane. PMID- 19551098 TI - Transmission behaviors of single mode hollow metallic waveguides dedicated to mid infrared nulling interferometry. AB - This paper reports the characterization of hollow metallic waveguides (HMW) to be used as single-mode wavefront filters for nulling interferometry in the 6 20microm range. The measurements presented here were performed using both single mode and multimode conductive waveguides at 10.6microm. We found propagation losses of about 16dB/mm, which are mainly due to the theoretical skin effect absorption in addition to the roughness of the waveguide's metallic walls. The input and output coupling efficiency of our samples has been improved by adding tapers to minimize the impedance mismatch. A proper distinction between propagation losses and coupling losses is presented. Despite their elevate propagation losses, HMW show excellent spatial filtering capabilities in a spectral range where photonics technologies are only emerging. PMID- 19551099 TI - Orthonormal vector polynomials in a unit circle, Part I: Basis set derived from gradients of Zernike polynomials. AB - Zernike polynomials provide a well known, orthogonal set of scalar functions over a circular domain, and are commonly used to represent wavefront phase or surface irregularity. A related set of orthogonal functions is given here which represent vector quantities, such as mapping distortion or wavefront gradient. These functions are generated from gradients of Zernike polynomials, made orthonormal using the Gram- Schmidt technique. This set provides a complete basis for representing vector fields that can be defined as a gradient of some scalar function. It is then efficient to transform from the coefficients of the vector functions to the scalar Zernike polynomials that represent the function whose gradient was fit. These new vector functions have immediate application for fitting data from a Shack-Hartmann wavefront sensor or for fitting mapping distortion for optical testing. A subsequent paper gives an additional set of vector functions consisting only of rotational terms with zero divergence. The two sets together provide a complete basis that can represent all vector distributions in a circular domain. PMID- 19551100 TI - Complete polarization state control of ultrafast laser pulses with a single linear spatial light modulator. AB - Shaping of the phase, amplitude, and polarization state of an ultrashort pulse is demonstrated using a novel arrangement of a single, linear, high-resolution liquid crystal array. Orthogonal polarization components, separated by a Wollaston prism, are manipulated independently and re-combined in a near-common path, common-optic geometry. PMID- 19551101 TI - Detection of multiple scattering in optical coherence tomography using the spatial distribution of Stokes vectors. AB - Multiple scattering is one of the main degrading influences in optical coherence tomography, but to date its presence in an image can only be indirectly inferred. We present a polarization-sensitive method that shows the potential to detect it more directly, based on the degree to which the detected polarization state at any given image point is correlated with the mean state over the surrounding region. We report the validation of the method in microsphere suspensions, showing a strong dependence of the degree of correlation upon the extent to which multiply scattered light is coherently detected. We demonstrate the method's utility in various tissues, including chicken breast ex vivo and human skin and nailfold in vivo. PMID- 19551102 TI - Bidirectional subwavelength slit splitter for THz surface plasmons. AB - We have conducted a feasibility study of a frequency splitter operating at THz frequencies, based on a bidirectional subwavelength slit simulated using two dimensional finite difference time domain (FDTD) techniques. The near-field wave emanating from the narrow slit serves as a subwavelength-scaled excitation source. By placing two optimized grating structures on the opposite sides of the slit, the THz waves at different frequencies are guided in the two desired directions. Confinement of the optical field is illustrated for different surface structures. PMID- 19551103 TI - Angle-resolved annular data acquisition method for microellipsometry. AB - An ellipsometric data acquisition method is introduced to measure the optical properties of sample. It is based on a microellipsometer hardware layout integrated a high numerical aperture objective lens, which is aligned in the normal direction of sample surface. This technique enables to achieve ellipsometric data at multiple incident angle with a sub-mum probe beam size, moreover real-time measurement is possible due to no moving parts. The experimental results of different SiO(2) thin film are demonstrated, also calibration technique is described. PMID- 19551104 TI - Microconcentrators to recover fill-factor in image photodetectors with pixel on board processing circuits. AB - We propose an array of non-imaging micro-concentrators as a mean to recover the loss of sensitivity due to area fill-factor. This is particularly important for those image photo detectors in which complex circuit functions are required and a substantial fraction of the pixel area is consumed, like e.g., 3D camera, SPAD arrays, fluorescence analyzers, etc., but also in CMOS sensors. So far, the low fill-factor was an unacceptable loss of sensitivity precluding from the development of such devices, whereas by using a concentrator array a recovery is possible, up to the inverse square of numerical aperture of the objective lens. By ray tracing, we calculate the concentration factors of several geometries of non-imaging concentrator, i.e., truncated cone, parabolic and compound parabolic, both reflective and refractive. The feasibility of a sizeable recovery of fill factor (up to 50) is demonstrated. PMID- 19551105 TI - Reconstruction of optical absorption coefficient maps of heterogeneous media by photoacoustic tomography coupled with diffusion equation based regularized Newton method. AB - We describe a novel reconstruction method that allows for quantitative recovery of optical absorption coefficient maps of heterogeneous media using tomographic photoacoustic measurements. Images of optical absorption coefficient are obtained from a diffusion equation based regularized Newton method where the absorbed energy density distribution from conventional photoacoustic tomography serves as the measured field data. We experimentally demonstrate this new method using tissue-mimicking phantom measurements and simulations. The reconstruction results show that the optical absorption coefficient images obtained are quantitative in terms of the shape, size, location and optical property values of the heterogeneities examined. PMID- 19551106 TI - Lensless light focusing with the centric marine diatom Coscinodiscus walesii. AB - In this work, we report on the light focusing ability exploited by the microshell of a marine organism: the Coscinodiscus wailesii diatom. A 100 microm spot size of a red laser beam is narrowed up to less than 10 microm at a distance of 104 microm after the transmission through the regular geometry of the diatom structure, which thus acts as a microlens. Numerical simulations of the electromagnetic field propagation show a good qualitative agreement with the experimental results. The focusing effect is due to the superposition of the waves scattered by the holes present on the surface of the diatom valve. Very interesting applications in micro-optic devices are feasible due to the morphological and biological characteristic of these unicellular organisms. PMID- 19551107 TI - The finite element method as applied to the diffraction by an anisotropic grating. AB - The main goal of the method proposed in this paper is the numerical study of various kinds of anisotropic gratings deposited on isotropic substrates, without any constraint upon the diffractive pattern geometry or electromagnetic properties. To that end we propose a new FEM (Finite Element Method) formulation which rigorously deals with each infinite issue inherent to grating problems. As an example, 2D numerical experiments are presented in the cases of the diffraction of a plane wave by an anisotropic aragonite grating on silica substrate (for the two polarization cases and at normal or oblique incidence). We emphasize the interesting property that the diffracted field is non symmetric in a geometrically symmetric configuration. PMID- 19551108 TI - Towards non-sequential double ionization of Ne and Ar using a femtosecond laser oscillator. AB - We report on first proof-of-principles results on non-sequential double ionization of argon and neon achieved by using a newly developed long-cavity Ti:sapphire femtosecond oscillator with a pulse duration of 45 fs and a repetition of 6.2 MHz combined with a dedicated reaction microscope. Under optimized experimental conditions, peak intensities larger than 2.310(14) W/cm(2) have been achieved. Ion momentum distributions were recorded for both rare gases and show significantly different features for single as well as for double ionization. For single ionization of neon a spike of zero-momentum electrons is found when decreasing the laser intensity towards the lowest ionization rate we can measure which is attributed to a non-resonant ionization channel. As to double ionization, the longitudinal momentum distribution for Ne(2+) displays a clear double-hump structure whereas this feature is found to be smoothened out with a maximum at zero momentum for Ar(2+). PMID- 19551109 TI - Point spread function of a collinear holographic storage system. AB - A paraxial solution to the coaxial holographic storage algorithm is proposed based on the scalar diffraction theory and a VOHIL model (volume hologram being an integrator of the lights emitted from elementary light sources), which can give insight into the optical characteristics of the collinear holographic storage system in an effective way, including the point spread function and shift selectivity. The paraxial solution shows that the reference pattern is the key issue in the point spread function. Thus, the bit error rate of the system can be improved by changing the reference pattern. The proposed solution will be useful in the design of a new reference pattern to perform a high-quality readout pattern in the coaxial holographic storage system. PMID- 19551110 TI - Tailoring the sensing capabilities of nanohole arrays in gold films with Rayleigh anomaly-surface plasmon polaritons. AB - Surface plasmon polaritons (SPPs) and diffraction effects such as Rayleigh anomalies (RAs) play key roles in the transmission of light through periodic subwavelength hole arrays in metal films. Using a combination of theory and experiment we show how refractive index (RI) sensitive transmission features arise from hole arrays in thin gold films. We show that large transmission amplitude changes occur over a narrow range of RI values due to coupling between RAs and SPPs on opposite sides of the metal film. Furthermore, we show how to predict, on the basis of a relatively simple analysis, the periodicity and other system parameters that should be used to achieve this "RA-SPP" effect for any desired RI range. PMID- 19551111 TI - Two-axis magnetically-driven MEMS scanning catheter for endoscopic high-speed optical coherence tomography. AB - A two-axis scanning catheter was developed for 3D endoscopic imaging with spectral domain optical coherence tomography (SD-OCT). The catheter incorporates a micro-mirror scanner implemented with microelectromechanical systems (MEMS) technology: the micro-mirror is mounted on a two-axis gimbal comprised of folded flexure hinges and is actuated by magnetic field. The scanner can run either statically in both axes or at the resonant frequency (>= 350Hz) for the fast axis. The assembled catheter has an outer diameter of 2.8 mm and a rigid part of 12 mm in length. Its scanning range is +/- 20 in optical angle in both axes with low voltages (1 approximately 3V), resulting in a scannable length of approximately 1 mm at the surface in both axes, even with the small catheter size. The catheter was incorporated with a multi-functional SD-OCT system for 3D endoscopic imaging. Both intensity and polarization-sensitive images could be acquired simultaneously at 18.5K axial scans/s. In vivo 3D images of human fingertips and oral cavity tissue are presented as a demonstration. PMID- 19551112 TI - Harmonically matched grating-based full-field quantitative high-resolution phase microscope for observing dynamics of transparent biological samples. AB - We have developed a full-field high resolution quantitative phase imaging technique for observing dynamics of transparent biological samples. By using a harmonically matched diffraction grating pair (600 and 1200 lines/mm), we were able to obtain non-trivial phase difference (other than 0 degrees or 180 degrees) between the output ports of the gratings. Improving upon our previous design, our current system mitigates astigmatism artifacts and is capable of high resolution imaging. This system also employs an improved phase extraction algorithm. The system has a lateral resolution of 1.6 mum and a phase sensitivity of 62 mrad. We employed the system to acquire high resolution phase images of onion skin cells and a phase movie of amoeba proteus in motion. PMID- 19551113 TI - Ultra-wideband pulse generation with flexible pulse shape and polarity control using a Sagnac-interferometer-based intensity modulator. AB - We propose a novel scheme to generate ultra-wideband (UWB) pulse by employing a Sagnac interferometer comprising a phase modulator. This structure performs a dual-input and dual-output intensity modulator (IM), ultimately resulting in the flexibility to select the shape and the polarity of the generated UWB pulse. The experiment results show a good agreement with the theoretical investigation in terms of both pulse profile and spectrum, which conforms to the definition of UWB signals by the U.S. Federal Communications Commission. Furthermore, the proposed scheme is independent of the voltage bias point. PMID- 19551114 TI - Laser operation of Yb3+ in disordered Li0.75Gd0.75Ba0.5(MoO4)2 crystal with small quantum defect. AB - A new strategy has been developed to enhance the optical bandwidths of rare earth dopants by partial substitution of the divalent cation (D) in the original DWO(4) or DMoO(4) crystal structures. For demonstration, the monoclinic (space group C2/c) Yb-doped Li(0.75)Gd(0.75)Ba(0.5)(MoO(4))(2) crystal was grown in a Li(2)Mo(2)O(7) flux, and 300 K Yb(3+) laser operation is reported. The laser emission is characterized by rather short wavelengths related to the specific features of the absorption and emission spectra, which leads to very small quantum defect, i.e., as low as 0.7% for E//b-axis. Using a Ti:sapphire laser at 976.6 nm, up to 295 mW of cw power are obtained without special cooling while the tunability range extends over 33 nm around 1020 nm. PMID- 19551115 TI - Heuristic Green's function of the time dependent radiative transfer equation for a semi-infinite medium. AB - The Green's function of the time dependent radiative transfer equation for the semi-infinite medium is derived for the first time by a heuristic approach based on the extrapolated boundary condition and on an almost exact solution for the infinite medium. Monte Carlo simulations performed both in the simple case of isotropic scattering and of an isotropic point-like source, and in the more realistic case of anisotropic scattering and pencil beam source, are used to validate the heuristic Green's function. Except for the very early times, the proposed solution has an excellent accuracy (> 98 % for the isotropic case, and > 97 % for the anisotropic case) significantly better than the diffusion equation. The use of this solution could be extremely useful in the biomedical optics field where it can be directly employed in conditions where the use of the diffusion equation is limited, e.g. small volume samples, high absorption and/or low scattering media, short source-receiver distances and early times. Also it represents a first step to derive tools for other geometries (e.g. slab and slab with inhomogeneities inside) of practical interest for noninvasive spectroscopy and diffuse optical imaging. Moreover the proposed solution can be useful to several research fields where the study of a transport process is fundamental. PMID- 19551116 TI - Multiaperture planar waveguide spectrometer formed by arrayed Mach-Zehnder interferometers. AB - Concept, theory and simulations of a new type of waveguide device, a multiaperture Fourier-transform planar waveguide spectrometer, are presented. The spectrometer is formed by an array of Mach-Zehnder interferometers generating a wavelength dependent spatial fringe pattern at the array output. The input light spectrum is calculated using a discrete Fourier transformation of the output spatial fringes. The multiaperture input significantly increases the optical throughput (etendue) compared to conventional single input spectrometers. Design rules for the arrayed spectrometer are deduced from performance specifications such as wavelength range and spectral resolution. A design example with spectral resolution 0.025 nm and range 2.5 nm is presented, where the optical throughput is increased by a factor of 200 compared to a single input device. PMID- 19551117 TI - Wavefront correction with a 37-actuator ferrofluid deformable mirror. AB - This paper discusses an innovative low-cost deformable mirror made of a magnetic liquid (ferrofluid) whose surface is actuated by an hexagonal array of small current carrying coils. Predicted and experimental performances of a 37-actuator ferrofluid deformable mirror are presented along with wavefront correction examples. We show the validity of the model used to compute the actuators currents to obtain a desired wavefront shape. We demonstrate that the ferrofluid deformable mirror can correct a 11 microm low order aberrated wavefront to a residual RMS error of 0.05 microm corresponding to a Strehl ratio of 0.82. PMID- 19551118 TI - Volume integral equation analysis of surface plasmon resonance of nanoparticles. AB - The interactions between electromagnetic field and arbitrarily shaped metallic nanoparticles are numerically investigated. The scattering and near field intensity of nanoparticles are characterized by using volume integral equation which is formulated by considering the total electric field, i.e. the sum of incident fields and radiated fields by equivalent electric volume currents, within the scatterers. The resultant volume integral equation is then discretized using divergence-conforming vector basis functions and is subsequently solved numerically. Numerical examples are presented to demonstrate the application of volume integral equation to capture and analyze the surface plasmon resonance of arbitrarily shaped metallic nanoparticles. The effects of illumination angles and background media to the surface plasmon resonance are also investigated. The results show that our proposed method is particularly useful and accurate in characterizing the surface plasmon properties of metallic nanoparticles. PMID- 19551119 TI - Adaptive optics for enhanced signal in CARS microscopy. AB - We report the use of adaptive optics with coherent anti-Stokes Raman scattering (CARS) microscopy for label-free deep tissue imaging based on molecular vibrational spectroscopy. The setup employs a deformable membrane mirror and a random search optimization algorithm to improve signal intensity and image quality at large sample depths. We demonstrate the ability to correct for both system and sample-induced aberrations in test samples as well as in muscle tissue in order to enhance the CARS signal. The combined system and sample-induced aberration correction increased the signal by an average factor of approximately 3x for the test samples at a depth of 700 microm and approximately 6x for muscle tissue at a depth of 260 microm. The enhanced signal and higher penetration depth offered by adaptive optics will augment CARS microscopy as an in vivo and in situ biomedical imaging modality. PMID- 19551120 TI - Picosecond-resolution fluorescence lifetime imaging microscopy: a useful tool for sensing molecular interactions in vivo via FRET. AB - Fluorescence lifetime imaging microscopy (FLIM) provides a promising, robust method of detecting molecular interaction not not nots in vivo via fluorescence/Forster resonance energy transfer (FRET), by monitoring the variation of donor fluorescence lifetime, which is insensitive to many artifacts influencing convential intensity-based measurements, e.g. fluorophore concentration, photobleaching, and spectral bleed-through. As proof of principle, we demonstrate the capability of a novel picosecond-resolution FLIM system to detect molecular interactions in a well-established FRET assay. We then apply the FLIM system to detect the molecular interaction of a transforming oncogene RhoC with a binding partner RhoGDIgamma in vivo, which is critical to understand and interfere with Rho signaling for cancer therapeutics. PMID- 19551121 TI - The quest for three-color entanglement: experimental investigation of new multipartite quantum correlations. AB - We experimentally investigate quadrature correlations between pump, signal, and idler fields in an above-threshold optical parametric oscillator. We observe new quantum correlations among the pump and signal or idler beams, as well as among the pump and a combined quadrature of signal and idler beams. A further investigation of unforeseen classical noise observed in this system is presented, which hinders the observation of the recently predicted tripartite entanglement. In spite of this noise, current results approach the limit required to demonstrate three-color entanglement. PMID- 19551122 TI - Suppression of XPM and XPM-induced nonlinear phase noise for RZ-DPSK signals in 40 Gbit/s WDM transmission systems with optimum dispersion mapping. AB - We numerically investigate XPM effect and XPM-induced nonlinear phase noise in both RZ-DPSK and multi-format (RZ-DPSK and RZ-OOK) WDM systems operating at 40 Gbit/s with different dispersion maps. The relative strength of XPM effect and XPM-induced nonlinear phase noise is discussed for both RZ-DPSK and multi-format WDM transmission. With optimum dispersion mapping, XPM and XPM-induced nonlinear phase noise from neighboring channels carrying either OOK or DPSK signals can both be effectively suppressed. PMID- 19551123 TI - Point light source integral imaging with improved resolution and viewing angle by the use of electrically movable pinhole array. AB - A resolution and viewing-angle enhanced integral imaging system using electrically movable pinhole array is proposed. A pinhole array on liquid crystal is adopted as dynamic pinhole array in integral imaging. The location of the pinhole array is controlled electrically. The pinhole array is expected to be moved fast enough to make an after-image effect, and the corresponding elemental images are displayed synchronously without reducing the 3D viewing aspect of the reconstructed image. With the proposed technique, the resolution and the viewing angle can be improved remarkably, and the upper resolution limit imposed by the Nyquist sampling theorem is overcome. The explanation of the proposed system is provided and the experimental results are also presented. PMID- 19551124 TI - The effect of external forces on discrete motion within holographic optical tweezers. AB - Holographic optical tweezers is a widely used technique to manipulate the individual positions of optically trapped micron-sized particles in a sample. The trap positions are changed by updating the holographic image displayed on a spatial light modulator. The updating process takes a finite time, resulting in a temporary decrease of the intensity, and thus the stiffness, of the optical trap. We have investigated this change in trap stiffness during the updating process by studying the motion of an optically trapped particle in a fluid flow. We found a highly nonlinear behavior of the change in trap stiffness vs. changes in step size. For step sizes up to approximately 300 nm the trap stiffness is decreasing. Above 300 nm the change in trap stiffness remains constant for all step sizes up to one particle radius. This information is crucial for optical force measurements using holographic optical tweezers. PMID- 19551125 TI - Characterizing and tracking single colloidal particles with video holographic microscopy. AB - We use digital holographic microscopy and Mie scattering theory to simultaneously characterize and track individual colloidal particles. Each holographic snapshot provides enough information to measure a colloidal sphere's radius and refractive index to within 1%, and simultaneously to measure its three-dimensional position with nanometer in-plane precision and 10 nanometer axial resolution. PMID- 19551126 TI - Fabrication and characterization of three-dimensional copper metallodielectric photonic crystals. AB - Three-dimensional metallodielectric photonic crystals were created by fabricating a micron-scale polymeric template using multiphoton direct laser writing (DLW) in SU-8 and conformally and selectively coating the template with copper (Cu) via nanoparticle-nucleated electroless metallization. This process deposits a uniform metal coating, even deep within a lattice, because it is not directional like sputter-coating or evaporative deposition. Infrared reflectance spectra show that upon metallization the optical behavior transitions fully from a dielectric photonic crystal to that of a metal photonic crystal (MPC). After depositing 50 nm of Cu, the MPCs exhibit a strong plasmonic stop band having reflectance greater than 80% across the measured part of the band and reaching as high as 95% at some wavelengths. Numerical simulations match remarkably well with the experimental data and predict all dominant features observed in the reflectance measurements, showing that the MPCs are structurally well formed. These data show that the Cu-based process can be used to create high performance MPCs and devices that are difficult or impossible to fabricate by other means. PMID- 19551127 TI - Ultra-broadband optical parametric generation and simultaneous RGB generation in periodically poled lithium niobate. AB - We report on efficient collinear optical parametric generation (OPG) with gain band ranging from 1400 to 2600 nm in a 2 cm-long periodically poled lithium niobate (PPLN) crystal. Such an ultra-broad gain band was obtained by choosing the pump wavelength at 933 nm, at which the group-velocities of the signal and the idler match near the degeneracy point. High OPG efficiency was obtained by quasi-phase matching (QPM). The ultra-broadband OPG led to efficient collinear RGB generation from a single PPLN crystal at a fixed pump wavelength. The green and red beams were found to be originating from high-order QPM sum-frequency generation between the pump and selected frequencies in the OPG band, while the blue beam was high-order QPM second-harmonic generation of the pump. PMID- 19551128 TI - Reconstruction for free-space fluorescence tomography using a novel hybrid adaptive finite element algorithm. AB - With the development of in-vivo free-space fluorescence molecular imaging and multi-modality imaging for small animals, there is a need for new reconstruction methods for real animal-shape models with a large dataset. In this paper we are reporting a novel hybrid adaptive finite element algorithm for fluorescence tomography reconstruction, based on a linear scheme. Two different inversion strategies (Conjugate Gradient and Landweber iterations) are separately applied to the first mesh level and the succeeding levels. The new algorithm was validated by numerical simulations of a 3-D mouse atlas, based on the latest free space setup of fluorescence tomography with 360 degrees geometry projections. The reconstructed results suggest that we are able to achieve high computational efficiency and spatial resolution for models with irregular shape and inhomogeneous optical properties. PMID- 19551129 TI - Point-by-point inscription of first-order fiber Bragg grating for C-band applications. AB - The influence of the fiber geometry on the point-by-point inscription of fiber Bragg gratings using a femtosecond laser is highlighted. Fiber Bragg gratings with high spectral quality and strong first-order Bragg resonances within the C band are achieved by optimizing the inscription process. Large birefringence (1.2x10(-4)) and high degree of polarization-dependent index modulation are observed in these gratings. Potential applications of these gratings in resonators are further illustrated. PMID- 19551130 TI - Elliptically modulated self-trapped singular beams in nonlocal nonlinear media: ellipticons. AB - We introduce a new class of elliptically modulated self-trapped singular beams in isotropic nonlinear media where nonlocality plays a crucial role in their existence. The analytical expressions in the highly nonlocal nonlinear limit of these elliptically shaped self-trapped beams, or ellipticons, is obtained and their existence in more general nonlocal nonlinear media is demonstrated. We show that the ellipticons represent a generalization of several known self-trapped beams, for example vortex solitons, azimuthons, and the Hermite and Laguerre solitons clusters. For the limit of the highly nonlocal nonlinear medium, the ellipticons are described in close form in terms of the InceGauss functions. PMID- 19551131 TI - Quantum state tomography of a fiber-based source of polarization-entangled photon pairs. AB - We report an experimental demonstration of a bright high-fidelity single-mode optical-fiber source of polarization-entangled photon pairs. The source takes advantage of single-mode fiber optics, highly nonlinear microstructure fiber, judicious phase-matching, and the inherent stability provided by a Sagnac interferometer. With a modest average pump power (300 microW), we create all four Bell states with a detected two-photon coincidence rate of 7 kHz per bandwidth of 0.9 nm, in a spectral range of more than 20 nm. To characterize the purity of the states produced by this source, we use quantum-state tomography to reconstruct the corresponding density matrices, with fidelities of 95 % or more for each Bell state. PMID- 19551132 TI - 2.7 W tunable orange-red GaInNAs semiconductor disk laser. AB - We report on a GaInNAs/GaAs semiconductor disk laser frequency-doubled to produce orange-red radiation. The disk laser operates at a fundamental wavelength of 1224 nm and delivers an output power of 2.68 W in the visible region with an optical to-optical conversion efficiency of 7.4%. The frequency-converted signal could be launched into a singlemode optical fiber with 70-78% coupling efficiency, demonstrating good beam quality for the visible radiation. Using a Fabry-Perot glass etalon the emission wavelength could be tuned over an 8 nm spectral range. PMID- 19551133 TI - Low-loss, flat-passband and athermal arrayed-waveguide grating multi/demultiplexer. AB - We successfully demonstrated a low-loss, flat-passband, and athermal arrayed waveguide grating (AWG) multi/demultiplexer with a Mach-Zehnder interferometer (MZI) as an input router. Resin-filled trenches were formed in the longer arm of the MZI as well as the slab in the AWG to compensate for the temperature dependence. A 32-channel athermal multi/demultiplexer was fabricated using silica based planar lightwave circuit (PLC) technology. A small temperature-dependent wavelength shift of 0.02 nm was obtained over the temperature range of -5 to 65 degrees C with low-loss (3.3-3.7 dB) and flat-passband spectra. PMID- 19551134 TI - Simulation and analysis of OOK-to-BPSK format conversion based on gain transparent SOA used as optical phase-modulator. AB - All-optical on-off keying (OOK) to binary phase-shift keying (BPSK) modulation format conversion based on gain-transparent semiconductor optical amplifier (GT SOA) is simulated and analyzed, where GT-SOA is used as an all-optical phase modulator (PM). Numerical simulation of the phase modulation effect of GT-SOA is performed using a wideband dynamic model of GT-SOA and the quality of the BPSK signal is evaluated using the differential-phase-Q factor. Performance improvement by holding light injection is analyzed and non-return-to-zero (NRZ) and return-to-zero (RZ) modulation formats of the OOK signal are considered. PMID- 19551135 TI - Induction of empathy by the smell of anxiety. AB - The communication of stress/anxiety between conspecifics through chemosensory signals has been documented in many vertebrates and invertebrates. Here, we investigate how chemosensory anxiety signals conveyed by the sweat of humans (N = 49) awaiting an academic examination are processed by the human brain, as compared to chemosensory control signals obtained from the same sweat donors in a sport condition. The chemosensory stimuli were pooled according to the donation condition and administered to 28 participants (14 males) synchronously to breathing via an olfactometer. The stimuli were perceived with a low intensity and accordingly only about half of the odor presentations were detected by the participants. The fMRI results (event-related design) show that chemosensory anxiety signals activate brain areas involved in the processing of social emotional stimuli (fusiform gyrus), and in the regulation of empathic feelings (insula, precuneus, cingulate cortex). In addition, neuronal activity within attentional (thalamus, dorsomedial prefrontal cortex) and emotional (cerebellum, vermis) control systems were observed. The chemosensory perception of human anxiety seems to automatically recruit empathy-related resources. Even though the participants could not attentively differentiate the chemosensory stimuli, emotional contagion seems to be effectively mediated by the olfactory system. PMID- 19551137 TI - Reverse engineering of gene regulatory networks: a comparative study. AB - Reverse engineering of gene regulatory networks has been an intensively studied topic in bioinformatics since it constitutes an intermediate step from explorative to causative gene expression analysis. Many methods have been proposed through recent years leading to a wide range of mathematical approaches. In practice, different mathematical approaches will generate different resulting network structures, thus, it is very important for users to assess the performance of these algorithms. We have conducted a comparative study with six different reverse engineering methods, including relevance networks, neural networks, and Bayesian networks. Our approach consists of the generation of defined benchmark data, the analysis of these data with the different methods, and the assessment of algorithmic performances by statistical analyses. Performance was judged by network size and noise levels. The results of the comparative study highlight the neural network approach as best performing method among those under study. PMID- 19551136 TI - Melanopsin bistability: a fly's eye technology in the human retina. AB - In addition to rods and cones, the human retina contains light-sensitive ganglion cells that express melanopsin, a photopigment with signal transduction mechanisms similar to that of invertebrate rhabdomeric photopigments (IRP). Like fly rhodopsins, melanopsin acts as a dual-state photosensitive flip-flop in which light drives both phototransduction responses and chromophore photoregeneration that bestows independence from the retinoid cycle required by rods and cones to regenerate photoresponsiveness following bleaching by light. To explore the hypothesis that melanopsin in humans expresses the properties of a bistable photopigment in vivo we used the pupillary light reflex (PLR) as a tool but with methods designed to study invertebrate photoreceptors. We show that the pupil only attains a fully stabilized state of constriction after several minutes of light exposure, a feature that is consistent with typical IRP photoequilibrium spectra. We further demonstrate that previous exposure to long wavelength light increases, while short wavelength light decreases the amplitude of pupil constriction, a fundamental property of IRP difference spectra. Modelling these responses to invertebrate photopigment templates yields two putative spectra for the underlying R and M photopigment states with peaks at 481 nm and 587 nm respectively. Furthermore, this bistable mechanism may confer a novel form of "photic memory" since information of prior light conditions is retained and shapes subsequent responses to light. These results suggest that the human retina exploits fly-like photoreceptive mechanisms that are potentially important for the modulation of non-visual responses to light and highlights the ubiquitous nature of photoswitchable photosensors across living organisms. PMID- 19551138 TI - CCHCR1 is up-regulated in skin cancer and associated with EGFR expression. AB - Despite chronic inflammation, psoriatic lesions hardly ever progress to skin cancer. Aberrant function of the CCHCR1 gene (Coiled-Coil alpha-Helical Rod protein 1, HCR) within the PSORS1 locus may contribute to the onset of psoriasis. As CCHCR1 is expressed in certain cancers and regulates keratinocyte (KC) proliferation in a transgenic mouse model, we studied its relation to proliferation in cutaneous squamous cell cancer (SCC) cell lines by expression arrays and quantitative RT-PCR and in skin tumors by immunohistochemistry. CCHCR1 protein was detected in the pushing border of SCC and lining basal cell carcinoma islands. Different from psoriasis, Ki67 had a similar expression pattern as CCHCR1. The most intense CCHCR1 staining occurred in areas positive for epidermal growth factor receptor (EGFR). Expression of CCHCR1 mRNA was upregulated 30-80% in SCC lines when compared to normal KCs and correlated positively with Ki67 expression. The most aggressive and invasive tumor cell lines (RT3, FaDu) expressed CCHCR1 mRNA less than non-tumorigenic HaCaT cells. Moreover, the tumor promoters okadaic acid and menadione downregulated CCHCR1 mRNA. We conclude that both in psoriasis and the early stages of KC transformation, CCHCR1 may function as a negative regulator of proliferation, but beyond a certain point in oncogenesis cannot control this phenomenon any longer. PMID- 19551139 TI - Cardiomyocyte contractile dysfunction in the APPswe/PS1dE9 mouse model of Alzheimer's disease. AB - OBJECTIVES: Ample clinical and experimental evidence indicated that patients with Alzheimer's disease display a high incidence of cardiovascular events. This study was designed to examine myocardial histology, cardiomyocyte shortening, intracellular Ca(2+) homeostasis and regulatory proteins, electrocardiogram, adrenergic response, endoplasmic reticulum (ER) stress and protein carbonyl formation in C57 wild-type (WT) mice and an APPswe/PS1dE9 transgenic (APP/PS1) model for Alzheimer's disease. METHODS: Cardiomyocyte mechanical properties were evaluated including peak shortening (PS), time-to-PS (TPS), time-to-relengthening (TR), maximal velocity of shortening and relengthening (+/-dL/dt), intracellular Ca(2+) transient rise and decay. RESULTS: Little histological changes were observed in APP/PS1 myocardium. Cardiomyocytes from APP/PS1 but not APP or PS1 single mutation mice exhibited depressed PS, reduced+/-dL/dt, normal TPS and TR compared with WT mice(.) Rise in intracellular Ca(2+) was lower accompanied by unchanged resting/peak intracellular Ca(2+) levels and intracellular Ca(2+) decay in APP/PS1 mice. Cardiomyocytes from APP/PS1 mice exhibited a steeper decline in PS at high frequencies. The responsiveness to adrenergic agonists was dampened although beta(1)-adrenergic receptor expression was unchanged in APP/PS1 hearts. Expression of the Ca(2+) regulatory protein phospholamban and protein carbonyl formation were downregulated and elevated, respectively, associated with unchanged SERCA2a, Na(+)-Ca(2+) exchanger and ER stress markers in APP/PS1 hearts. Our further study revealed that antioxidant N-acetylcysteine attenuated the contractile dysfunction in APP/PS1 mice. CONCLUSIONS: Our results depicted overt cardiomyocyte mechanical dysfunction in the APP/PS1 Alzheimer's disease model, possibly due to oxidative stress. PMID- 19551140 TI - Simultaneous live cell imaging using dual FRET sensors with a single excitation light. AB - Fluorescence resonance energy transfer (FRET) between fluorescent proteins is a powerful tool for visualization of signal transduction in living cells, and recently, some strategies for imaging of dual FRET pairs in a single cell have been reported. However, these necessitate alteration of excitation light between two different wavelengths to avoid the spectral overlap, resulting in sequential detection with a lag time. Thus, to follow fast signal dynamics or signal changes in highly motile cells, a single-excitation dual-FRET method should be required. Here we reported this by using four-color imaging with a single excitation light and subsequent linear unmixing to distinguish fluorescent proteins. We constructed new FRET sensors with Sapphire/RFP to combine with CFP/YFP, and accomplished simultaneous imaging of cAMP and cGMP in single cells. We confirmed that signal amplitude of our dual FRET measurement is comparable to of conventional single FRET measurement. Finally, we demonstrated to monitor both intracellular Ca(2+) and cAMP in highly motile cardiac myocytes. To cancel out artifacts caused by the movement of the cell, this method expands the applicability of the combined use of dual FRET sensors for cell samples with high motility. PMID- 19551141 TI - Analysis of germline variants in CDH1, IGFBP3, MMP1, MMP3, STK15 and VEGF in familial and sporadic renal cell carcinoma. AB - BACKGROUND: The investigation of rare familial forms of kidney cancer has provided important insights into the biology of sporadic renal cell carcinoma (RCC). In particular, the identification of the von Hippel Lindau (VHL) familial cancer syndrome gene (VHL) provided the basis for the discovery that VHL is somatically inactivated in most sporadic clear cell RCC. Many cases of familial RCC do not have mutations in known RCC susceptibility genes and there is evidence that genetic modifiers may influence the risk of RCC in VHL disease patients. Hence we hypothesised that low-penetrance functional genetic variants in pathways related to the VHL protein (pVHL) function might (a) modify the phenotypic expression of VHL disease and/or (b) predispose to sporadic RCC. METHODOLOGY/PRINCIPAL FINDINGS: We tested this hypothesis for functional polymorphisms in CDH1 (rs16260), IGFBP3 (rs2854744), MMP1 (rs1799750), MMP3 (rs679620), STK15 (rs2273535) and VEGF (rs1570360). We observed that variants of MMP1 and MMP3 were significant modifiers of RCC risk (and risks of retinal angioma and cerebellar haemangioblastoma) in VHL disease patients. In addition, higher frequencies of the MMP1 rs1799750 2G allele (p = 0.017, OR 1.49, 95%CI 1.06-2.08) and the MMP1/MMP3 rs1799750/rs679620 2G/G haplotype (OR 1.45, 95%CI 1.01-2.10) were detected in sporadic RCC patients than in controls (n = 295). CONCLUSIONS/SIGNIFICANCE: These findings (a) represent the first example of genetic modifiers of RCC risk in VHL disease, (b) replicate a previous report of an association between MMP1/MMP3 variants and sporadic RCC and (c) further implicate MMP1/MMP3-related pathways in the pathogenesis of familial and sporadic RCC. PMID- 19551142 TI - No need to discriminate? Reproductive diploid males in a parasitoid with complementary sex determination. AB - Diploid males in hymenopterans are generally either inviable or sterile, thus imposing a severe genetic load on populations. In species with the widespread single locus complementary sex determination (sl-CSD), sex depends on the genotype at one single locus with multiple alleles. Haploid (hemizygous) individuals are always males. Diploid individuals develop into females when heterozygous and into males when homozygous at the sex determining locus. Our comparison of the mating and reproductive success of haploid and diploid males revealed that diploid males of the braconid parasitoid Cotesia glomerata sire viable and fertile diploid daughters. Females mated to diploid males, however, produced fewer daughters than females mated to haploid males. Nevertheless, females did not discriminate against diploid males as mating partners. Diploid males initiated courtship display sooner than haploid males and were larger in body size. Although in most species so far examined diploid males were recognized as genetic dead ends, we present a second example of a species with sl-CSD and commonly occurring functionally reproductive diploid males. Our study suggests that functionally reproductive diploid males might not be as rare as hitherto assumed. We argue that the frequent occurrence of inbreeding in combination with imperfect behavioural adaptations towards its avoidance promote the evolution of diploid male fertility. PMID- 19551143 TI - A fish eye out of water: ten visual opsins in the four-eyed fish, Anableps anableps. AB - The "four-eyed" fish Anableps anableps has numerous morphological adaptations that enable above and below-water vision. Here, as the first step in our efforts to identify molecular adaptations for aerial and aquatic vision in this species, we describe the A. anableps visual opsin repertoire. We used PCR, cloning, and sequencing to survey cDNA using unique primers designed to amplify eight sequences from five visual opsin gene subfamilies, SWS1, SWS2, RH1, RH2, and LWS. We also used Southern blotting to count opsin loci in genomic DNA digested with EcoR1 and BamH1. Phylogenetic analyses confirmed the identity of all opsin sequences and allowed us to map gene duplication and divergence events onto a tree of teleost fish. Each of the gene-specific primer sets produced an amplicon from cDNA, indicating that A. anableps possessed and expressed at least eight opsin genes. A second PCR-based survey of genomic and cDNA uncovered two additional LWS genes. Thus, A. anableps has at least ten visual opsins and all but one were expressed in the eyes of the single adult surveyed. Among these ten visual opsins, two have key site haplotypes not found in other fish. Of particular interest is the A. anableps-specific opsin in the LWS subfamily, S180gamma, with a SHYAA five key site haplotype. Although A. anableps has a visual opsin gene repertoire similar to that found in other fishes in the suborder Cyprinodontoidei, the LWS opsin subfamily has two loci not found in close relatives, including one with a key site haplotype not found in any other fish species. A. anableps opsin sequence data will be used to design in situ probes allowing us to test the hypothesis that opsin gene expression differs in the distinct ventral and dorsal retinas found in this species. PMID- 19551144 TI - Modulation of the intestinal microbiota alters colitis-associated colorectal cancer susceptibility. AB - It is well established that the intestinal microbiota plays a key role in the pathogenesis of Crohn's disease (CD) and ulcerative colitis (UC) collectively referred to as inflammatory bowel disease (IBD). Epidemiological studies have provided strong evidence that IBD patients bear increased risk for the development of colorectal cancer (CRC). However, the impact of the microbiota on the development of colitis-associated cancer (CAC) remains largely unknown. In this study, we established a new model of CAC using azoxymethane (AOM)-exposed, conventionalized-Il10(-/-) mice and have explored the contribution of the host intestinal microbiota and MyD88 signaling to the development of CAC. We show that 8/13 (62%) of AOM-Il10(-/-) mice developed colon tumors compared to only 3/15 (20%) of AOM- wild-type (WT) mice. Conventionalized AOM-Il10(-/-) mice developed spontaneous colitis and colorectal carcinomas while AOM-WT mice were colitis-free and developed only rare adenomas. Importantly, tumor multiplicity directly correlated with the presence of colitis. Il10(-/-) mice mono-associated with the mildly colitogenic bacterium Bacteroides vulgatus displayed significantly reduced colitis and colorectal tumor multiplicity compared to Il10(-/-) mice. Germ-free AOM-treated Il10(-/-) mice showed normal colon histology and were devoid of tumors. Il10(-/-); Myd88(-/-) mice treated with AOM displayed reduced expression of Il12p40 and Tnfalpha mRNA and showed no signs of tumor development. We present the first direct demonstration that manipulation of the intestinal microbiota alters the development of CAC. The TLR/MyD88 pathway is essential for microbiota induced development of CAC. Unlike findings obtained using the AOM/DSS model, we demonstrate that the severity of chronic colitis directly correlates to colorectal tumor development and that bacterial-induced inflammation drives progression from adenoma to invasive carcinoma. PMID- 19551145 TI - Raf Kinase Inhibitory Protein protects cells against locostatin-mediated inhibition of migration. AB - BACKGROUND: Raf Kinase Inhibitory Protein (RKIP, also PEBP1), a member of the Phosphatidylethanolamine Binding Protein family, negatively regulates growth factor signaling by the Raf/MAP kinase pathway. Since an organic compound, locostatin, was reported to bind RKIP and inhibit cell migration by a Raf dependent mechanism, we addressed the role of RKIP in locostatin function. METHODS/FINDINGS: We analyzed locostatin interaction with RKIP and examined the biological consequences of locostatin binding on RKIP function. NMR studies show that a locostatin precursor binds to the conserved phosphatidylethanolamine binding pocket of RKIP. However, drug binding to the pocket does not prevent RKIP association with its inhibitory target, Raf-1, nor affect RKIP phosphorylation by Protein Kinase C at a regulatory site. Similarly, exposure of wild type, RKIP depleted HeLa cells or RKIP-deficient (RKIP(-/-)) mouse embryonic fibroblasts (MEFs) to locostatin has no effect on MAP kinase activation. Locostatin treatment of wild type MEFs causes inhibition of cell migration following wounding. RKIP deficiency impairs migration further, indicating that RKIP protects cells against locostatin-mediated inhibition of migration. Locostatin treatment of depleted or RKIP(-/-) MEFs reveals cytoskeletal disruption and microtubule abnormalities in the spindle. CONCLUSIONS/SIGNIFICANCE: These results suggest that locostatin's effects on cytoskeletal structure and migration are caused through mechanisms independent of its binding to RKIP and Raf/MAP kinase signaling. The protective effect of RKIP against drug inhibition of migration suggests a new role for RKIP in potentially sequestering toxic compounds that may have deleterious effects on cells. PMID- 19551146 TI - Stimulus familiarity affects perceptual restoration in the European starling (Sturnus vulgaris). AB - BACKGROUND: Humans can easily restore a speech signal that is temporally masked by an interfering sound (e.g., a cough masking parts of a word in a conversation), and listeners have the illusion that the speech continues through the interfering sound. This perceptual restoration for human speech is affected by prior experience. Here we provide evidence for perceptual restoration in complex vocalizations of a songbird that are acquired by vocal learning in a similar way as humans learn their language. METHODOLOGY/PRINCIPAL FINDINGS: European starlings were trained in a same/different paradigm to report salient differences between successive sounds. The birds' response latency for discriminating between a stimulus pair is an indicator for the salience of the difference, and these latencies can be used to evaluate perceptual distances using multi-dimensional scaling. For familiar motifs the birds showed a large perceptual distance if discriminating between song motifs that were muted for brief time periods and complete motifs. If the muted periods were filled with noise, the perceptual distance was reduced. For unfamiliar motifs no such difference was observed. CONCLUSIONS/SIGNIFICANCE: The results suggest that starlings are able to perceptually restore partly masked sounds and, similarly to humans, rely on prior experience. They may be a suitable model to study the mechanism underlying experience-dependent perceptual restoration. PMID- 19551147 TI - Neuron-specific regulation of associative learning and memory by MAGI-1 in C. elegans. AB - BACKGROUND: Identifying the molecular mechanisms and neural circuits that control learning and memory are major challenges in neuroscience. Mammalian MAGI/S-SCAM is a multi-PDZ domain synaptic scaffolding protein that interacts with a number of postsynaptic signaling proteins and is thereby thought to regulate synaptic plasticity [1], [2], [3]. PRINCIPAL FINDINGS: While investigating the behavioral defects of C. elegans nematodes carrying a mutation in the single MAGI ortholog magi-1, we have identified specific neurons that require MAGI-1 function for different aspects of associative learning and memory. Various sensory stimuli and a food deprivation signal are associated in RIA interneurons during learning, while additional expression of MAGI-1 in glutamatergic AVA, AVD and possibly AVE interneurons is required for efficient memory consolidation, i.e. the ability to retain the conditioned changes in behavior over time. During associative learning, MAGI-1 in RIA neurons controls in a cell non-autonomous fashion the dynamic remodeling of AVA, AVD and AVE synapses containing the ionotropic glutamate receptor (iGluR) GLR-1 [4]. During memory consolidation, however, MAGI 1 controls GLR-1 clustering in AVA and AVD interneurons cell-autonomously and depends on the ability to interact with the beta-catenin HMP-2. SIGNIFICANCE: Together, these results indicate that different aspects of associative learning and memory in C. elegans are likely carried out by distinct subsets of interneurons. The synaptic scaffolding protein MAGI-1 plays a critical role in these processes in part by regulating the clustering of iGluRs at synapses. PMID- 19551148 TI - Large-scale assessment of the effect of popularity on the reliability of research. AB - Based on theoretical reasoning it has been suggested that the reliability of findings published in the scientific literature decreases with the popularity of a research field. Here we provide empirical support for this prediction. We evaluate published statements on protein interactions with data from high throughput experiments. We find evidence for two distinctive effects. First, with increasing popularity of the interaction partners, individual statements in the literature become more erroneous. Second, the overall evidence on an interaction becomes increasingly distorted by multiple independent testing. We therefore argue that for increasing the reliability of research it is essential to assess the negative effects of popularity and develop approaches to diminish these effects. PMID- 19551149 TI - Suppression of glomerulonephritis in NZB/NZW lupus prone mice by adoptive transfer of ex vivo expanded regulatory T cells. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease of unknown cause characterized by expansion of autoreactive lymphocytes. Regulatory T cells (T(regs)) are a component of the normal immune system and contribute to the maintenance of peripheral tolerance. T(reg) abnormalities have been associated with several autoimmune diseases and there is interest in the role of T(regs) in SLE. We previously demonstrated that transfer of expanded CD4(+)CD25(+)CD62L(HI) T(regs) slows the development of lupus in (NZBxNZW)F(1) (B/W) mice. However in the absence of T(reg) specific surface antigens, cell purification remains a compromise between the breadth and purity of the population isolated. Importantly, purified populations always contain Foxp3(-) effector T cells (T(effs)) that theoretically could exacerbate autoimmunity in the recipient. Here we explore the impact of transferring the more comprehensive, but less pure T(reg) subset defined by CD4(+)CD25(+) expression on development of murine lupus. All cells were FACS sorted and expanded prior to adoptive transfer. Development of proteinuria and survival were measured. We found that exogenous expansion of CD4(+)CD25(+) cells produced a population containing 70-85% CD4(+)Foxp3(+)T(regs). Expanded T(regs) had higher CTLA-4 and Foxp3 expression, increased in vitro suppression capacity, and prolonged in vivo survival as compared to freshly isolated cells. Adoptive transfer of expanded CD4(+)CD25(+) T(regs) inhibited the onset of glomerulonephritis and prolonged survival in mice. Importantly the population of T(eff) contained within the adoptively transferred cells had reduced survival and proliferation capacity as compared to either co transferred T(regs) or transferred T(effs) expanded in the absence of T(regs). These studies demonstrate that adoptive transfer of expanded CD4(+)CD25(+)Foxp3(+)T(regs) has the capacity to inhibit the onset of murine lupus and that this capacity is significant despite transfer of co-cultured T(eff) cells. These data indicate that when co-expanded with regulatory T cells, exogenously activated T(effs) from autoimmune patients may not pose a significant risk of promoting disease. PMID- 19551150 TI - Epstein-Barr virus independent dysregulation of UBP43 expression alters interferon-stimulated gene expression in Burkitt lymphoma. AB - Epstein-Barr virus (EBV) persists as a life-long latent infection within memory B cells, but how EBV may circumvent the innate immune response within this virus reservoir is unclear. Recent studies suggest that the latency-associated non coding RNAs of EBV may actually induce type I (antiviral) interferon production, raising the question of how EBV counters the negative consequences this is likely to have on viral persistence. We addressed this by examining the type I interferon response in Burkitt lymphoma (BL) cell lines, the only in vitro model of the restricted program of EBV latency-gene expression in persistently infected B cells in vivo. Importantly, we observed no effect of EBV on interferon alpha induced signaling or evidence of type I interferon production, suggesting that EBV in this latent state is silent to the cell's innate antiviral surveillance. We did uncover, however, a defect in the negative feedback control of interferon signaling in a subpopulation of BL lines as was revealed by prolonged interferon stimulated gene transcription consistent with sustained tyrosine phosphorylation on STAT1 and STAT2. This was due to inadequate induction of expression of the ubiquitin-specific protease UBP43, which removes the ubiquitin-like ISG15 polypeptide conjugated to proteins (ISGylation) in response to type I interferons. Results here are consistent with previous findings in genetically engineered Ubp43(-/-) murine cells that UBP43 down-regulates interferon signaling, independent of its ISG15 isopeptidase activity, by precluding the protein kinase JAK1 from the interferon receptor. This natural deficiency in UBP43 expression may therefore provide a useful model to further probe the biological roles of UBP43 and ISGylation. PMID- 19551151 TI - MYC is a metastasis gene for non-small-cell lung cancer. AB - BACKGROUND: Metastasis is a process by which cancer cells learn to form satellite tumors in distant organs and represents the principle cause of death of patients with solid tumors. NSCLC is the most lethal human cancer due to its high rate of metastasis. METHODOLOGY/PRINCIPAL FINDINGS: Lack of a suitable animal model has so far hampered analysis of metastatic progression. We have examined c-MYC for its ability to induce metastasis in a C-RAF-driven mouse model for non-small-cell lung cancer. c-MYC alone induced frank tumor growth only after long latency at which time secondary mutations in K-Ras or LKB1 were detected reminiscent of human NSCLC. Combination with C-RAF led to immediate acceleration of tumor growth, conversion to papillary epithelial cells and angiogenic switch induction. Moreover, addition of c-MYC was sufficient to induce macrometastasis in liver and lymph nodes with short latency associated with lineage switch events. Thus we have generated the first conditional model for metastasis of NSCLC and identified a gene, c-MYC that is able to orchestrate all steps of this process. CONCLUSIONS/SIGNIFICANCE: Potential markers for detection of metastasis were identified and validated for diagnosis of human biopsies. These markers may represent targets for future therapeutic intervention as they include genes such as Gata4 that are exclusively expressed during lung development. PMID- 19551153 TI - Seizure (Ictal)--EEG characteristics in subgroups of depressive disorder in patients receiving electroconvulsive therapy (ECT)--a preliminary study and multivariate approach. AB - OBJECTIVES: Examine frequency distributions of ictal EEG after ECT stimulation in diagnostic subgroups of depression. METHODS: EEG registration was consecutively monitored in 33 patients after ECT stimulation. Patients were diagnosed according to DSM IV and subdivided into: (1) major depressive disorder with psychotic features (n = 7), (2) unipolar depression (n = 20), and (3) bipolar depression (n = 6). RESULTS: Results indicate that the diagnostically subgroups differ in their ictal EEG frequency spectrumml: (1) psychotic depression has a high occurrence of delta and theta waves, (2) unipolar depression has high occurrence of delta, theta and gamma waves, and (3) bipolar depression has a high occurrence of gamma waves. A linear discriminant function separated the three clinical groups with an accuracy of 94%. CONCLUSION: Psychotic depressed patients differ from bipolar depression in their frequency based on probability distribution of ictal EEG. Psychotic depressed patients show more prominent slowing of EEG than nonpsychotic depressed patients. Thus the EEG results may be supportive in classifying subgroups of depression already at the start of the ECT treatment. PMID- 19551152 TI - Methodologies for in vitro cloning of small RNAs and application for plant genome(s). AB - The "RNA revolution" that started at the end of the 20th century with the discovery of post-transcriptional gene silencing and its mechanism via RNA interference (RNAi) placed tiny 21-24 nucleotide long noncoding RNAs (ncRNAs) in the forefront of biology as one of the most important regulatory elements in a host of physiologic processes. The discovery of new classes of ncRNAs including endogenous small interfering RNAs, microRNAs, and PIWI-interacting RNAs is a hallmark in the understanding of RNA-dependent gene regulation. New generation high-throughput sequencing technologies further accelerated the studies of this "tiny world" and provided their global characterization and validation in many biological systems with sequenced genomes. Nevertheless, for the many "yet unsequenced" plant genomes, the discovery of small RNA world requires in vitro cloning from purified cellular RNAs. Thus, reproducible methods for in vitro small RNA cloning are of paramount importance and will remain so into the foreseeable future. In this paper, we present a description of existing small RNA cloning methods as well as next-generation sequencing methods that have accelerated this research along with a description of the application of one in vitro cloning method in an initial small RNA survey in the "still unsequenced" allotetraploid cotton genome. PMID- 19551154 TI - Computational challenges in miRNA target predictions: to be or not to be a true target? AB - All microRNA (miRNA) target--finder algorithms return lists of candidate target genes. How valid is that output in a biological setting? Transcriptome analysis has proven to be a useful approach to determine mRNA targets. Time course mRNA microarray experiments may reliably identify downregulated genes in response to overexpression of specific miRNA. The approach may miss some miRNA targets that are principally downregulated at the protein level. However, the high-throughput capacity of the assay makes it an effective tool to rapidly identify a large number of promising miRNA targets. Finally, loss and gain of function miRNA genetics have the clear potential of being critical in evaluating the biological relevance of thousands of target genes predicted by bioinformatic studies and to test the degree to which miRNA-mediated regulation of any "validated" target functionally matters to the animal or plant. PMID- 19551155 TI - [Not Available]. AB - The cyclin-dependent kinase (Cdk)-cyclin D/retinoblastoma (pRb)/E2F cascade, which controls the G1/S transition of cell cycle, has been found to be altered in many neoplasias. Inhibition of this pathway by using, for example, selective Cdk4 inhibitors has been suggested to be a promising approach for cancer therapy. We hypothesized that appropriately radiolabeled Cdk4 inhibitors are suitable probes for tumor imaging and may be helpful studying cell proliferation processes in vivo by positron emission tomography. Herein, we report the synthesis and biological, biochemical, and radiopharmacological characterizations of two (124)I labeled small molecule Cdk4 inhibitors (8-cyclopentyl-6-iodo-5-methyl-2-(4 piperazin-1-yl-phenylamino)-8H-pyrido[2,3-d]-pyrimidin-7-one (CKIA) and 8 cyclopentyl-6-iodo-5-methyl-2-(5-(piperazin-1-yl)-pyridin-2-yl-amino)-8H pyrido[2,3-d]pyrimidin-7-one (CKIB)). Our data demonstrate a defined and specific inhibition of tumor cell proliferation through CKIA and CKIB by inhibition of the Cdk4/pRb/E2F pathway emphasizing potential therapeutic benefit of CKIA and CKIB. Furthermore, radiopharmacological properties of [(124)I]CKIA and [(124)I]CKIB observed in human tumor cells are promising prerequisites for in vivo biodistribution and imaging studies. PMID- 19551157 TI - The levels of tumor necrosis factor-alpha and interleukin-6 in patients with isolated coronary artery ectasia. AB - BACKGROUND/AIM: Coronary artery ectasia (CAE) is considered as a variant of atherosclerosis. Tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) are among the sensitive markers of systemic inflammation. The aim of this study was to evaluate the plasma levels of the cytokines; TNF-alpha and IL-6 in CAE patients. METHODS: Plasma concentrations of TNF-alpha and IL-6 were measured in 36 patients with CAE (28 males, mean age: 58.2 +/- 12 years), and results were compared with age and sex-matched controls (n = 32) without coronary artery ectasia. TNF-alpha and IL-6 concentrations in blood were assessed by enzyme linked immunosorbent assay (ELISA). RESULTS: Baseline characteristics of the two groups were similar. TNF-alpha and IL-6 levels were significantly higher in CAE group than controls (15.6 +/- 11.2 pg/mL versus 7.8 +/- 3.7 pg/mL, P < .001, and 17.2 +/- 12.6 versus 7.6 +/- 2.1 P < .0001, resp.). CONCLUSION: CAE patients showed increases in TNF-alpha and IL-6 levels compared to the controls. This study provides evidence for alterations in the proinflammatory cytokines which suggest the involvement of the immune system in the pathophysiology of CAE. Further placebo-controlled studies are needed to evaluate the clinical significance of this increase in TNF-alpha and IL-6 levels. PMID- 19551156 TI - Mesothelioma cells escape heat stress by upregulating Hsp40/Hsp70 expression via mitogen-activated protein kinases. AB - Therapy with hyperthermal chemotherapy in pleural diffuse malignant mesothelioma had limited benefits for patients. Here we investigated the effect of heat stress on heat shock proteins (HSP), which rescue tumour cells from apoptosis. In human mesothelioma and mesothelial cells heat stress (39-42 degrees C) induced the phosphorylation of two mitogen activated kinases (MAPK) Erk1/2 and p38, and increased Hsp40, and Hsp70 expression. Mesothelioma cells expressed more Hsp40 and were less sensitive to heat stress compared to mesothelial cells. Inhibition of Erk1/2 MAPK by PD98059 or by Erk1 siRNA down-regulated heat stress-induced Hsp40 and Hsp70 expression and reduced mesothelioma cell survival. Inhibition of p38MAPK by SB203580 or siRNA reduced Hsp40, but not Hsp70, expression and also increased mesothelioma cell death. Thus hyperthermia combined with suppression of p38 MAPK or Hsp40 may represent a novel approach to improve mesothelioma therapy. PMID- 19551159 TI - Resonant Mode-hopping Micromixing. AB - A common micromixer design strategy is to generate interleaved flow topologies to enhance diffusion. However, problems with these designs include complicated structures and dead volumes within the flow fields. We present an active micromixer using a resonating piezoceramic/silicon composite diaphragm to generate acoustic streaming flow topologies. Circulation patterns are observed experimentally and correlate to the resonant mode shapes of the diaphragm. The dead volumes in the flow field are eliminated by rapidly switching from one discrete resonant mode to another (i.e., resonant mode-hop). Mixer performance is characterized by mixing buffer with a fluorescence tracer containing fluorescein. Movies of the mixing process are analyzed by converting fluorescent images to two dimensional fluorescein concentration distributions. The results demonstrate that mode-hopping operation rapidly homogenized chamber contents, circumventing diffusion-isolated zones. PMID- 19551158 TI - Heterologous expression of a membrane-spanning auxin importer: implications for functional analyses of auxin transporters. AB - Biochemical studies of plant auxin transporters in vivo are made difficult by the presence of multiple auxin transporters and auxin-interacting proteins. Furthermore, the expression level of most such transporters in plants is likely to be too low for purification and downstream functional analysis. Heterologous expression systems should address both of these issues. We have examined a number of such systems for their efficiency in expressing AUX1 from Arabidopsis thaliana. We find that a eukaryotic system based upon infection of insect cells with recombinant baculovirus provides a high level, easily scalable expression system capable of delivering a functional assay for AUX1. Furthermore, a transient transfection system in mammalian cells enables localization of AUX1 and AUX1-mediated transport of auxin to be investigated. In contrast, we were unable to utilise P. pastoris or L. lactis expression systems to reliably express AUX1. PMID- 19551160 TI - Fundamental studies of molecular depth profiling and 3D imaging using Langmuir Blodgett films as a model. AB - Molecular depth profiling and three-dimensional imaging using cluster projectiles and SIMS have become a prominent tool for organic and biological materials characterization. To further explore the fundamental features of cluster bombardment of organic materials, especially depth resolution and differential sputtering, we have developed a reproducible and robust model system consisting of Langmuir-Blodgett (LB) multilayer films. Molecular depth profiles were acquired, using a 40-keV C(60) (+) probe, with LB films chemically alternating between barium arachidate and barium dimyristoyl phosphatidate. The chemical structures were successfully resolved as a function of depth. The molecular ion signals were better preserved when the experiment was performed under cryogenic conditions than at room temperature. A novel method was used to convert the scale of fluence into depth which facilitated quantitative measurement of the interface width. Furthermore, the LB films were imaged as a function of depth. The reconstruction of the SIMS images correctly represented the original chemical structure of the film. It also provided useful information about interface mixing and edge effects during sputtering. PMID- 19551161 TI - Carbon-Centered Free Radicals in Particulate Matter Emissions from Wood and Coal Combustion. AB - Electron paramagnetic resonance (EPR) spectroscopy was used to measure the free radicals in the particulate matter (PM) emissions from wood and coal combustion. The intensity of radicals in PM dropped linearly within two months of sample storage and stabilized after that. This factor of storage time was adjusted when comparing radical intensities among different PM samples. An inverse relationship between coal rank and free radical intensities in PM emissions was observed, which was in contrast with the pattern of radical intensities in the source coals. The strong correlation between intensities of free radical and elemental carbon in PM emissions suggests that the radical species may be carbon-centered. The increased g-factors, 2.0029-2.0039, over that of purely carbon-centered radicals may indicate the presence of vicinal oxygen heteroatom. The redox and biology activities of these carbon-centered radicals are worthy of evaluation. PMID- 19551165 TI - 2D-IR spectroscopy: ultrafast insights into biomolecule structure and function. AB - Ultrafast 2D-IR spectroscopy is rapidly becoming a valuable tool for examining the relationship between structure and function of biomolecules. The unique combination of molecular-level structural information and ultrafast time resolution gives previously inaccessible insights relating to the rapid structural fluctuations, vibrational dynamics and solvent-solute interactions of proteins, all of which have important implications for the biological function of these species. In this tutorial review, the method and development of ultrafast 2D-IR spectroscopy is discussed, including an introduction to the two main experimental approaches, double resonance and photon echo 2D-IR, and the extension of the technique to non-equilibrium or transient 2D-IR measurements. The scope of the new information available through 2D-IR spectroscopy is then demonstrated by reference to the current state of the art of 2D-IR studies of molecules of biological interest. PMID- 19551166 TI - Rational design and applications of highly efficient reaction systems photocatalyzed by noble metal nanoparticle-loaded titanium(IV) dioxide. AB - The fundamentals and applications of TiO(2) photocatalysis have been extensively studied in recent decades, the central theme being increasing reaction efficiency. A guideline for achieving so-called "reasonable delivery photocatalytic reaction systems (RDPRSs)" has been presented. This tutorial review summarizes recent developments in the RDPRS-based design of highly efficient photocatalytic reactions by noble metal nanoparticle-loaded TiO(2) (M/TiO(2)). After introducing the conditions required for RDPRSs, the key factors affecting the photocatalytic activity, including Fermi energy in the photostationary state, the metal particle size effect, and solvent effects are discussed. We then describe selected applications of RDPRSs for the conversion of light energy to chemical energy, environmental purification, and low-temperature cleaning of sulfur-poisoned metal catalysts. Future applications of this idea to visible-light photocatalysts, which are currently the subject of intensive research, may lead to feasible green and efficient photocatalytic reaction systems using sunlight as an energy source. PMID- 19551167 TI - Fluorescence modulation with photochromic switches in nanostructured constructs. AB - This tutorial review illustrates the structural design, photochemical and photophysical properties of nanostructured constructs incorporating luminescent and photochromic components. In these systems, the pronounced structural and electronic modifications that accompany the transformations of the photochromic components can be exploited to modulate the emission intensity of the luminescent components on the basis of electron and energy transfer processes. These photoresponsive systems can be assembled by: (1) integrating fluorescent and photochromic components within the main chain of the same polymer; (2) attaching multiple photochromes to a fluorescent organic polymer or luminescent inorganic nanoparticle; (3) appending either independent fluorophores and photochromes or fluorophore-photochrome dyads to a common polymer scaffold; (4) trapping distinct fluorophores and photochromes within the hydrophobic interior of the same cross linked polymer. In all instances, the changes in absorbance and/or redox potentials associated with the reversible interconversion of the two states of each photochromic component regulate the radiative deactivation of the luminescent components. As a result, the emission intensity of these nanoscaled assemblies can reversibly be switched between high and low values under the influence of optical stimulations. Thus, these clever operating principles for fluorescence modulation can lead to the development of innovative functional and nanostructured materials with photoresponsive character. In particular, protocols for the optical writing and reading of data as well as luminescent probes for bioimaging applications might ultimately emerge from these fundamental studies on photoresponsive molecular switches. PMID- 19551168 TI - Cationic inorganic materials for anionic pollutant trapping and catalysis. AB - Despite their vast industrial importance, almost all extended inorganic synthetic structures and naturally occurring minerals adopt a neutral or anionic charge. This tutorial review describes the current knowledge of inorganic materials that bear a positive charge on the host. The best-known example is the family of clays known as layered double hydroxides, and other limited examples also exist. Our research efforts have given rise to a series of new cationic inorganic materials, oxides and fluorides of heavy p-block metals. Organic-based cationic materials will also be briefly discussed. Interest in these materials is widespread, with potential applications in many areas including anionic pollutant trapping, exchange, selective catalysis, polymer hybrids and nanocomposites, and biomaterials for drug or DNA delivery. PMID- 19551169 TI - Mass spectrometry: from proteomics to metabolomics and lipidomics. AB - Mass spectrometry (MS) in combination with liquid chromatography (LC), i.e. LC MS, is the key analytical technique on which the emerging "omics" technologies of proteomics, metabolomics and lipidomics are based. It provides both structural and quantitative data and can be used in a "global" or "targeted" manner allowing on the one hand the identification of thousands of proteins from a tissue, or on the other the detection of biologically active metabolites at levels of a few parts-per-billion. It can be expected that the continued incremental developments in LC-MS along with data-handling routines will soon bear fruit in the quest for a greater understanding of human disease, leading to new drug targets and therapies. This tutorial review on proteomics, metabolomics and lipidomics will be of interest to the biochemical, biomedical and pharmaceutical communities, as well as those working in other branches of analytical bioscience. PMID- 19551170 TI - Carbon nanotubes: biomaterial applications. AB - There is increasing interest in the unique biological and medical properties of carbon nanotubes (CNTs), and it is expected that biomaterials incorporating CNTs will be developed for clinical use. There has been a great deal of progress in improving the various properties of CNTs for use in biomaterials and for promotion of tissue regeneration as scaffold materials. The effects of CNTs on cells and tissues are extremely important for their use in biomaterials. This tutorial review clarifies the current state of knowledge in the interdisciplinary field of CNT-based nanobiotechnology to determine whether CNTs may be useful in biomaterials. Future perspectives in this rapidly developing field will also be discussed. PMID- 19551171 TI - Silica-based chromogenic and fluorogenic hybrid chemosensor materials. AB - This tutorial review focuses on the recent development of silica-based organic inorganic hybrid nanomaterials for use in biological and environmental applications, in which these chromogenic and fluorogenic chemosensors can selectively detect and separate specific anions and neutral organic guests as well as toxic metal ions. PMID- 19551172 TI - trans-cyclopentane-1,2-diamine: the second youth of the forgotten diamine. AB - Despite its early description, trans-cyclopentane-1,2-diamine has been underestimated historically. Its non-commercial availability, extreme instability and complexity of the classical reported syntheses have produced far less interest amongst chemists than its higher homologue trans-cyclohexane-1,2 diamine, perhaps the most widely used diamine for the synthesis of ligands and receptors. However, the recent development of novel and efficient synthetic approaches has stimulated renewed interest in this chiral motif for a broad range of applications. In the first part of this tutorial review we shall discuss the existing methods for the preparation of trans-cyclopentane-1,2-diamine and some of its derivatives, in both racemic and enantioenriched forms. In subsequent sections, recent findings employing this diamine as a scaffold for chiral ligands, receptors and biologically active compounds will be highlighted. PMID- 19551173 TI - Techniques for the study and development of microbial fuel cells: an electrochemical perspective. AB - Microbial fuel cells (MFCs) represent a clean and renewable energy resource. To date, power generation in MFCs is severely limited. In order to improve performance, a wide range of techniques have been utilised for a fundamental scientific understanding of the components and processes and also to investigate MFC performance bottlenecks. In this tutorial review, we discuss the electrochemical/electroanalytical techniques employed in recent MFC studies and discusses the principles, experimental implementation, data processing requirements, capabilities, and weaknesses of these techniques. PMID- 19551174 TI - Catalytic asymmetric direct Mannich reaction: a powerful tool for the synthesis of alpha,beta-diamino acids. AB - Optically active alpha,beta-diamino acids are very attractive targets in organic synthesis because of their wide-ranging biological significance and high versatility as synthetic building blocks. Efficient synthesis of such non proteinogenic amino acid derivatives must face the challenge of generating two contiguous stereocenters with complete diastereo- and enantiocontrol in flexible, acyclic molecules. The catalytic asymmetric direct Mannich reaction has provided elegant and efficient solutions for the stereocontrolled assembly of both syn- and anti-alpha,beta-diamino acid derivatives, including those with a alpha tetrasubstituted carbon stereocenter, with the aid of either organometallic or purely organic chiral catalysts (or the combination of both). This tutorial review highlights progress in this area, which has recently been boosted through two complementary strategies: the direct Mannich reaction of glycine ester Schiff bases with imines and the direct aza-Henry reaction between nitro compounds and imines. PMID- 19551175 TI - Target-selective peptide-cleaving catalysts as a new paradigm in drug design. AB - This tutorial review describes the evolution of peptide-hydrolyzing metal catalysts towards artificial metalloproteases cleaving target proteins selectively. The catalytic cleavage of the backbone of a protein related to a disease may effect a cure. In particular, a new therapeutic option for amyloid diseases such as Alzheimer's disease, diabetes and Parkinson's disease has been presented. The new paradigm of drug design based on artificial metalloproteases should be of interest to researchers in the areas of biomimetic chemistry, as well as medicinal chemistry. PMID- 19551176 TI - Recent conceptual and technological advances in polydiacetylene-based supramolecular chemosensors. AB - Owing to the color (blue-to-red) and fluorescence (non-to-fluorescent) changes that take place in response to environmental perturbations, conjugated polydiacetylenes (PDAs) have been actively employed as sensory materials for the detection of biologically-, environmentally- and chemically-important target molecules. Until recently, the majority of PDA sensors have been prepared in the form of aqueous suspensions or Langmuir-type thin films on solid substrates. In order to overcome the limitations associated with conventional solution/film sensors, conceptually new formats, such as immobilized PDAs in and on solid substrates, microarrayed PDA sensors, microfluidic PDA sensors, as well as PDA embedded electrospun fiber sensors and resonance energy transfer (RET)-based PDA sensors, have been developed recently. In this tutorial review, the recent conceptual and technological achievements made in the area of conjugated PDA chemosensors are described. PMID- 19551178 TI - Solution combustion synthesis of oxide semiconductors for solar energy conversion and environmental remediation. AB - In this tutorial review, we summarize recent research on the solution combustion synthesis of oxide semiconductors for applications related to photovoltaic solar energy conversion, photoelectrochemical hydrogen generation, and heterogeneous photocatalytic remediation of environmental pollutants. First, the advantages of combustion synthesis relative to other strategies for preparing oxide semiconductors are discussed followed by a summary of process variants in combustion synthesis. The possibility of in situ chemical modification of the oxide during its formation in the combustion environment is addressed. Morphological and crystal structure aspects of the combustion-synthesized products are discussed followed by a summary of trends in their photocatalytic activity relative to benchmark samples prepared by other methods. PMID- 19551177 TI - The synergy of elemental and biomolecular mass spectrometry: new analytical strategies in life sciences. AB - The application of mass spectrometry with soft ionization techniques (ESI, electrospray ionization, and MALDI, matrix-assisted laser desorption ionization) in the life sciences for the detection and identification of biomolecules is already well established, whereas the application of elemental mass spectrometry and in particular inductively coupled plasma mass spectrometry (ICP-MS) for the determination of metals, metalloids and non-metals in biomolecules is rather new and there is some hesitation in accepting this analytical method, although it offers many advantages. Therefore, it is the aim of this tutorial review to highlight new analytical strategies consisting of the combined applications of elemental and molecular mass spectrometric techniques. In fact, elemental and biomolecular mass spectrometric methods are highly complementary: elemental mass spectrometry methods, such as ICP-MS, offer very sensitive element analysis in the trace and ultra-trace concentration range with multielement capability and the excellent and uniform sensitivity is structure-independent and can be used analytically for accurate quantification as well as for fast screening of specific elements even in complex samples. Laser ablation (LA) ICP-MS, as a solid state mass spectrometric technique, allows the direct determination of trace elements in biological and environmental samples and is applied for microlocal analysis with spatial resolution in the mum range. In contrast, molecular weight determination and structural information is completely lost during the ionization step so that these features have to be provided by biomolecular mass spectrometry and in particular by ESI- and MALDI-MS. On the basis of selected examples, it will be shown that only the combination of different elemental and biomolecular mass spectrometric techniques can solve analytical problems in the life sciences and environmental research in a synergistic way where neither technique alone would be successful. This synergy will be demonstrated by selected applications from various areas: food and nutrition, toxicology, clinical and pharmaceutical research, biochemistry and in particular proteomics. Future developments and trends will be discussed concerning instrumental developments of new mass spectrometric techniques providing high sensitivity with lower detection limits for many elements measured quasi-simultaneously so that new analytical information about biological systems can be drawn from isotopic information and the application of stable non-radioactive isotopic tracers. In addition, elemental labels enable the development of new high-throughput screening techniques based on multiplexed biomarkers. Advanced powerful surface mass spectrometric techniques are required for the imaging of elemental and molecular information in order to analyse tissue samples and to develop novel array-based biochips. PMID- 19551179 TI - Photocatalysis. A multi-faceted concept for green chemistry. AB - Photocatalysis (by semiconductors, molecules and ions) is used in such diverse applications as water hydrolysis for producing hydrogen as fuel, organic synthesis and the recovery of polluted effluents. This tutorial review discusses the common principles of such applications and their role in green chemistry. PMID- 19551180 TI - The chemical biology of modular biosynthetic enzymes. AB - Fatty acid synthase (FAS), polyketide synthase (PKS), and nonribosomal peptide synthetase (NRPS) modular biosynthetic enzymes are responsible for the production of a multitude of structurally diverse and biologically important small molecule natural products. Traditional biochemical and genetic studies of these enzymes have contributed substantially to the understanding of their underlying biosynthetic mechanisms. More recently these investigations have been aided by the skillful application of a combination of chemical and biological techniques to aid in overcoming the unique challenges associated with the enzymology of these large multifunctional enzymes. This critical review provides a historical context and details studies (through July 2008) which aim to identify and characterize these enzymes using synthetically and/or chemoenzymatically generated small molecule probes (233 references). PMID- 19551181 TI - Cellulose modification by polymer grafting: a review. AB - This critical review is concerned with the recent advances in graft polymerisation techniques involving cellulose and its derivatives. It summarises some of the features of cellulose structure and cellulose reactivity. Also described are the various techniques for grafting synthetic polymers from the cellulosic substrate. In addition to the traditional grafting techniques, we highlight the recent developments in polymer synthesis that allow increased control over the grafting process and permit the production of functional celluloses that possess improved physical properties and chemical properties (189 references). PMID- 19551182 TI - Advanced approach to polycyclics by a synergistic combination of enyne metathesis and Diels-Alder reaction. AB - Enyne metathesis (EM) has widely been used to prepare various synthetic and natural products. Further, a sequential use of EM and Diels-Alder (DA) reactions generates highly functionalized and intricate polycarbocycles and heterocyclic frameworks. In this critical review we describe the application of a unique combination of EM and DA reactions to prepare various amino acid derivatives, natural products and heterocycles. Some of the heterocyclic targets include cyclic peroxides, siloxanes and various nitrogen (or oxygen) containing compounds. Use of ethylene and cyclooctadiene (COD) for improving the yield of EM products are also discussed (69 references). PMID- 19551183 TI - Stereocontrolled routes to beta,beta'-disubstituted alpha-amino acids. AB - Owing to their significant abundance in natural products, chiral beta,beta' disubstituted alpha-amino acids remain an important synthetic objective. Emphasis has been focused in this critical review on the great diversity of enantio- and diastereoselective methodologies to reach these highly functionalized compounds. The oldest and cutting edge synthetic methods are described in parallel with the synthesis of many relevant biologically active targets (224 references). PMID- 19551184 TI - Calix[4]arene daisychains. AB - Generic calix[4]arenes became readily accessible in the late 70s. With their potential eight anchoring points, their utility for the production of sophisticated, highly functionalised macrocyclic molecules was rapidly recognised. While most studies in calixarene chemistry have focused on monocalixarene derivatives, there is now an increasing interest in developing multicalixarene compounds, especially those made of several linearly-arranged calix[4]arene units, the first examples of which were reported in 1989. This critical review will present the most important synthetic routes to such molecules together with an analysis of the properties that such cavity combinations may induce. In particular it will be shown that the nature of the links between the calixarene units plays a determinant role in the product properties and that singly-linked calixarenes can be exploited in varied applications, including those as efficient receptors of large molecules, as electrochemical and luminescent sensors in ion detection, or as new materials allowing capsule formation suitable for the storage of small guests (82 references). PMID- 19551188 TI - Theoretical evaluation of zeolite confinement effects on the reactivity of bulky intermediates. AB - Zeolites provide a unique setting for heterogeneous Bronsted acid catalysis, because the effects of the surrounding framework on fundamental reaction kinetics go well beyond what would be expected for a mere reaction flask. This aspect becomes very pronounced when bulky molecules form key intermediates for the reaction under study, which is exactly when the interaction between the framework and the intermediate is maximal. We will use the example of methanol-to-olefin conversion (MTO), and, more specifically, the constant interplay between the inorganic host framework and the organic hydrocarbon pool co-catalyst, to illustrate how zeolite confinement directly influences catalytic reaction rates. Theoretical calculations are used to isolate and quantify these specific effects, with the main focus on methylbenzenes in ZSM-5, as the archetypical MTO catalyst. This review intends to give an overview of recent theoretical insights, which have proven to provide an ideal complementary tool to experimental investigations. In addition, we will also introduce the role of zeolite breathing in activating a catalytic cycle. PMID- 19551189 TI - Initiated and oxidative chemical vapor deposition: a scalable method for conformal and functional polymer films on real substrates. AB - Chemical vapor deposition (CVD) is a widely-used technology for the preparation of conformal and defect-free inorganic thin films with systematically tunable properties. Polymers are a desirable class of materials for surface modification because of their low cost, wide array of chemical and physical functionality and mechanical flexibility. Initiated and oxidative chemical vapor deposition (iCVD and oCVD) are polymer CVD methods that combine the benefits of CVD processing with the possibilities of polymeric materials. Using these technologies, our laboratory has synthesized a number of functional, biocompatible and electrically conducting polymers as thin films on micro- and nano-structured surfaces. This Perspective will review recent advances in these areas and highlight devices and applications that utilize iCVD and oCVD polymers. PMID- 19551190 TI - IM-19: a new flexible microporous gallium based-MOF framework with pressure- and temperature-dependent openings. AB - Five metal-organic frameworks (MOFs) based on the same three-dimensional gallium terephthalate network (IM-19) are described, and an incommensurate structure (for the as-synthesized form) as well as two remarkable guest-free polymorphs (open and closed) are highlighted. PMID- 19551191 TI - Electronic charge transfer between ceria surfaces and gold adatoms: a GGA+U investigation. AB - We use density functional theory calculations with Hubbard corrections (DFT+U) to investigate electronic aspects of the interaction between ceria surfaces and gold atoms. Our results show that Au adatoms at the (111) surface of ceria can adopt Au(0), Au(+) or Au(-) electronic configurations depending on the adsorption site. The strongest adsorption sites are on top of the surface oxygen and in a bridge position between two surface oxygen atoms, and in both cases charge transfer from the gold atom to one of the Ce cations at the surface is involved. Adsorption at other sites, including the hollow sites of the surface, and an O-Ce bridging site, is weaker and does not involve charge transfer. Adsorption at an oxygen vacancy site is very strong and involves the formation of an Au(-) anion. We argue that the ability of gold atoms to stabilise oxygen vacancies at the ceria surface by moving into the vacancy site and attracting the excess electrons of the defect could be responsible for the enhanced reducibility of ceria surfaces in the presence of gold. Finally, we rationalise the differences in charge transfer behaviour from site to site in terms of the electrostatic potential at the surface and the coordination of the species. PMID- 19551192 TI - Alkali metals (Li, Na, and K) in methyl phosphodiester hydrolysis. AB - The phosphodiester linkage central to biological systems has been modeled by methyl phosphodiester (MPDE) in various theoretical and experimental studies. Under physiological conditions, hydrolysis of the phosphodiester is negligible, however this process can be catalyzed in the presence of metal ions. To understand the role of alkali metals in MPDE hydrolysis and, in particular, how it influences the reaction pathway and the associated energetics, density functional calculations employing the 6-31+G(d,p) basis set have been carried out. Different pathways that include the reactant, intermediates and the products have been investigated for MPDE hydrolysis catalyzed by one or two lithium ions, characterized as stationary point geometries on the potential energy surface. The pathways A and B incorporate a single lithium ion bonded to different oxygens of the diester functionality. In pathway C, a six-membered ring was noticed wherein the nucleophile bridges two lithium ions interacting with different oxygens of the phosphoryl group. Furthermore, in the pathway (D) incorporating two lithium ions, one of the lithium ions interacts with the hydroxyl group and another with the methoxy oxygen; both metal ions are coordinated by the same phosphoryl oxygen. In addition to this, yet another pathway (E), where the metal ions are bound to different oxygens of the phosphoryl group, has also been dealt with. The calculations have shown that the A and B pathways lead to a single step reaction. A three-step mechanism including the nucleophilic (hydroxyl) attack, rotation of a methyl group and, finally, departure of the methoxy group has been predicted for the D and E profiles. Both D and E pathways are favored equally (with a marginal difference of 0.3 kJ mol(-1) in their activation energies) in the gas phase and a transition state corresponding to nucleophilic attack with an energy barrier of 32.5 kJ mol(-1) was located when lithium was used. A penta-coordinated phosphorous intermediate on the potential energy surface was characterized along these pathways. MPDE hydrolysis yielded a lower energy barrier for lithium than those for the remaining alkali metal ions. This agrees well with the experimentally observed trend for the hydrolysis rates: Li > Na > K. Self consistent reaction field (SCRF) calculations reveal the lower energy barrier between the reactant and the transition state for the nucleophilic attack in nonpolar solvents. The extent of bond formation (or cleavage) in different stationary point structures along the reaction path as estimated from the electron density at the bond critical point in the molecular electron density topography, has proven useful in distinguishing the associative or dissociative reaction pathways. PMID- 19551193 TI - Dimerisation of nitrile oxides: a quantum-chemical study. AB - The [3 + 2] and [3 + 3] cyclodimerisation processes of small nitrile oxides, XCNO (X = F, Cl, Br, CN, CH(3)) are investigated by ab initio coupled cluster theory at the CCSD, CCSD(T) and MR-AQCC levels for the first time. The favoured dimerisation process is a multi-step reaction to furoxans (1,2,5-oxadiazole-2 oxides) involving dinitrosoalkene-like intermediates with diradical character. The rate determining step for all but the F-species is the first, corresponding to the C-C bond formation. The kinetic energy barrier depends on the nature of the substituent X, generally increasing with decreasing electronegativity and increasing pi-donor ability of the substituent: F (DeltaG(298) = 0 kJ mol(-1)) < Cl (72) < Br (90) < CH(3) (104) < CN (114) (MR-AQCC(2,2)//UB3LYP/cc-pVTZ). Following initial C-C bond formation, three possible dinitrosoethylene diradical pathways are explored. Two of them are new, and one of them is a low-energy three step path with implications for cycloreversion, tautomerism and detection of dinitrosoethylene intermediates. Alternative one-step, concerted [3 + 2] and [3 + 3] cyclodimerisation processes leading to 1,2,4-oxadiazole-4-oxides and 1,4,2,5 dioxadiazines have kinetic energy barriers around 100-240 kJ mol(-1) (CCSD//B3LYP), some 1.6 to 2.5 times higher than those leading to furoxans, supporting the experimental observations of furoxan formation as nitrile oxide loss channels during storage, trapping/re-vaporisation and reactions of nitrile oxides. Potential polymerisation initiation processes for NCCNO, involving the 1,2-dipolar NC substituent are also explored. PMID- 19551194 TI - Autocatalytic degradation of white phosphorus with silylenes. AB - White phosphorus (P(4)) is prone to undergo degradation by nucleophiles and is reluctant to do so with electrophiles. Silylenes possess a strong singlet character but at the same time bear a largely inert lone pair orbital at the silicon atom. Thus they predominantly react in a similar way to electrophilic carbenes. Due to the poor pi-character of the P-P bonds in white phosphorus, the overlap with the empty orbital for the electrophilic silylene is less facile and results in a relatively large barrier for the addition reaction. The electrophilic approach of the silylene to white phosphorus is catalyzed by addition of a second P(4), forming a trigonal bipyramidal transition state geometry. Its stability towards fragmentation is essentially lower than that of the silyl cation. The entropy contributions for bimolecular versus termolecular reactions are discussed. PMID- 19551195 TI - Symmetry breaking in spin-restricted Hartree-Fock solutions: the case of the C2 molecule and the N(2)+ and F(2)+ cations. AB - We examine a spin-preserving stability of restricted Hartree-Fock (RHF) and open shell RHF (ROHF) solutions for homonuclear diatomic species, namely the molecule C(2) and the N(2)(+) and F(2)(+)cations, in the entire relevant range of internuclear separations. In the presence of respective singlet, doublet, or triplet instabilities we explore the implied broken-symmetry (BS) solutions and check their stability. We also address the occurrence of vanishing roots rendered by the stability problem in the case of BS solutions. Since for homonuclear diatomic species the space symmetry of the nuclear framework cannot be broken, the only possible space symmetry breaking involves either the electron charge distribution or the rotational symmetry about the internuclear axis. Such BS solutions arise primarily due to the symmetry breaking of the valence, mostly frontier, molecular orbitals, which approach atomic-type orbitals in the dissociation limit. The resulting BS RHF or ROHF solutions yield much more realistic potential energy curves (PECs) than do the symmetry-adapted (SA) solutions. For the sake of comparison, we also generated corresponding PECs using the density functional theory (DFT). Finally, we examine the role of BS molecular orbitals in post-HF correlated approaches to the many-electron problem, specifically in the computation of equilibrium properties using the coupled cluster method with singles and doubles (CCSD) and its perturbatively corrected version for triples, the CCSD(T) method. PMID- 19551196 TI - Sequential or superexchange mechanism in bridged electron transfer distinguished by dynamics at a bridging molecule. AB - Two kinds of mechanisms are well known for electron transfer (ET) in the system DMA where a donor D and an acceptor A are fixed in a close distance by a bridging molecule M. When the free energy DeltaG(m) of the intermediate state |m of DM(-)A is much higher than the thermal energy k(B)T above the initial state of D(-)MA, the ET occurs unistep from D to A by the superexchange (SX) mechanism, passing |m as a quantum-mechanical virtual state. However, when DeltaG(m) becomes not much higher than k(B)T, the ordinary sequential (OS) ET may manifest itself, where the second ET from |m to the final state of DMA(-) takes place after thermalization of phonons in |m. Recently, much interest has been aroused in how the SX mechanism switches to the OS one as DeltaG(m) is lowered. This subject has often been described conventionally by summation of the rate constant for the SX mechanism and that for the OS one. However, such convention cannot be justified, since these mechanisms are realized in mutually opposite limits concerned with dynamics in mediation of ET by a bridging molecule, hence they cannot both be realized simultaneously in a single system. An observation of such a bridged ET by Paulson, Miller, Gan and Closs (J. Am. Chem. Soc. 2005, 127, 4860) provides a good example of this feature. Describing their observation in a unified framework for the ET, it is shown that the switch occurs at DeltaG(m) congruent with-0.5 eV, which is much lower than 0.3 eV reported by them, where the hot-sequential ET reveals itself, taking place during thermalization of phonons in |m. PMID- 19551197 TI - UV resonance Raman spectroscopy of TTR(105-115): determination of the pKa of tyrosine. AB - The 11-residue peptide fragment from transthyretin (TTR(105-115)) has been investigated using UV resonance Raman spectroscopy. Excitation at 239.5 nm reveals selective enhancement of scattering from two Tyr residues. The titrating behavior of the tyrosines is followed through the change in the Y8a band (1617 cm(-1)) frequency as a function of pH, and a pK(a) = 10.2 +/- 0.2 is obtained. This is compared to the value of 9.1 +/- 0.2 for the pK(a) of aqueous Tyr also obtained in the present study. The pK(a) difference observed here, along with observations in the nu(OH) region, suggest that the two Tyr residues in the peptide probe two distinct microenvironments. PMID- 19551198 TI - Assessment of the amide-I local modes in gamma- and beta-turns of peptides. AB - The amide-I local modes, mainly the C[double bond, length as m-dash]O stretching vibrations, form the structural basis of femtosecond 2D IR spectroscopy in characterizing backbone structures and dynamics of peptides and proteins. In this work, a density functional theory (DFT) level of computational assessment of the amide-I local modes in oligomers mostly in the turn conformations was carried out. It is shown that local mode properties, including transition frequencies and transition dipole magnitudes and orientations, are slightly conformational dependent. However, the distributions of these properties in the peptide oligomers are narrow and have mean values almost identical to those from an isolated peptide monomer, justifying the prevalent use of a uniform local mode in modeling the 1D and 2D IR spectra. In addition, it is shown that the transition dipole magnitude and orientation of the peptide monomer predicted by the DFT calculations can be well approximated by electrostatic potential-based transition charge schemes, e.g. Merz-Singh-Kollman, CHELP, as well as CHELPG. PMID- 19551199 TI - Reactions in gas phase and condensed phase C6F5X (X = NCO, CH2CN) triggered by low energy electrons. AB - Electron attachment to gas phase perfluorophenylisocyanate (C(6)F(5)NCO) and perfluorophenyloacetonitrile (C(6)F(5)CH(2)CN) generates metastable parent anions within a very narrow resonance close to zero energy. At higher energies (2-7 eV), dissociative electron attachment (DEA) resonances are present, associated with the rupture of the C(6)F(5)-X bond (X = NCO, CH(2)CN) with the excess electron finally localised on either of the two fragments. The most intense fragment ion from C(6)F(5)CH(2)CN (M) is (M - HF)(-), which arises from the loss of a neutral HF from the transient anion and requires the concerted cleavage of two bonds and formation of a new molecule (HF). Most remarkably, this rather complex DEA reaction is by about two orders of magnitude more intense than the single bond cleavages (C(6)F(5)-X) leading to the complementary DEA reactions C(6)F(5) + X(-) and C(6)F(5)(-) + X. From both condensed molecules we observe desorption of F(-) and CN(-) and, additionally, O(-) from C(6)F(5)NCO. The desorption yields also show a resonant behaviour with the peak maxima in the range 8-12 eV, i.e., near or above the ionization energy, indicating that in electron stimulated desorption (ESD) highly excited resonances are involved. Ab initio calculations are performed in order to get information on the shape and energy of the molecular orbitals involved in low energy (<2 eV) electron attachment. PMID- 19551200 TI - The gas-phase reaction between silylene and 2-butyne: kinetics, isotope studies, pressure dependence studies and quantum chemical calculations. AB - Time-resolved kinetic studies of the reactions of silylene, SiH(2), and dideutero silylene, SiD(2), generated by laser flash photolysis of phenylsilane and phenylsilane-d(3), respectively, have been carried out to obtain rate coefficients for their bimolecular reactions with 2-butyne, CH(3)C[triple bond, length as m-dash]CCH(3). The reactions were studied in the gas phase over the pressure range 1-100 Torr in SF(6) bath gas at five temperatures in the range 294 612 K. The second-order rate coefficients, obtained by extrapolation to the high pressure limits at each temperature, fitted the Arrhenius equations where the error limits are single standard deviations: log(k(H)(Infinity)/cm(3) molecule( 1) s(-1) = (-9.67 +/- 0.04) + (1.71 +/- 0.33) kJ mol(1)/RTIn10log(k(D)(Infinity)/cm(3) molecule(-1) s(-1) = (-9.65 +/- 0.01) + (1.92 +/- 0.13) kJ mol(-1)/RTIn10. Additionally, pressure-dependent rate coefficients for the reaction of SiH(2) with 2-butyne in the presence of He (1 100 Torr) were obtained at 301, 429 and 613 K. Quantum chemical (ab initio) calculations of the SiC(4)H(8) reaction system at the G3 level support the formation of 2,3-dimethylsilirene [cyclo-SiH(2)C(CH(3))[double bond, length as m dash]C(CH(3))-] as the sole end product. However, reversible formation of 2,3 dimethylvinylsilylene [CH(3)CH[double bond, length as m-dash]C(CH(3))SiH] is also an important process. The calculations also indicate the probable involvement of several other intermediates, and possible products. RRKM calculations are in reasonable agreement with the pressure dependences at an enthalpy value for 2,3 dimethylsilirene fairly close to that suggested by the ab initio calculations. The experimental isotope effects deviate significantly from those predicted by RRKM theory. The differences can be explained by an isotopic scrambling mechanism, involving H-D exchange between the hydrogens of the methyl groups and the D-atoms in the ring in 2,3-dimethylsilirene-1,1-d(2). A detailed mechanism involving several intermediate species, which is consistent with the G3 energy surface, is proposed to account for this. PMID- 19551201 TI - Study of the CH3 H2O radical complex stabilized in helium nanodroplets. AB - The weakly bound CH(3)H(2)O radical complex has been investigated by infrared laser spectroscopy. The complex is stabilized in helium nanodroplets and prepared by sequential pick up of a methyl radical and water molecule. Partially rotationally resolved spectra corresponding to the v = 1 <-- 0 excitation of the symmetric H(2)O stretching vibration within the complex show a significant red shift (25.06 cm(-1)) when compared with the symmetric stretch of H(2)O monomer, in agreement with the hydrogen bonded like structure derived by theory. Additional broad features were observed in the region predicted by theory for the antisymmetric stretch supporting our assignment. The B rotational constant is found to be 3.03 times smaller than predicted by ab initio calculations, with the reduction being attributed to the effects of helium solvation. The permanent electric dipole moment of the complex is experimentally determined to be 2.1 +/- 0.3 D using Stark spectroscopy. Ab initio calculations are also reported that provide support to the experimental results, as well as investigate the nature of large amplitude vibrational motion within the complex. PMID- 19551202 TI - Excited-state lifetime of propadienylidene, l-C3H2. AB - The excited-state dynamics of the singlet carbene propadienylidene, l-C(3)H(2), were investigated by femtosecond time-resolved photoionisation. The carbene was excited into the C (1)A(1) state with 250 nm pulses and the subsequent excited state dynamics were probed by multiphoton ionization with 800 nm pulses. The lifetime of the C (1)A(1) state was determined to be 70 fs. In agreement with recent nanosecond experiments, we assume that the carbene deactivates to the electronic ground state where it subsequently dissociates. Since propadienylidene was generated from 3-bromo-1-iodopropyne, two further radical intermediates were studied, IC(3)H(2) and C(3)H(2)Br. For both species, an ultrafast excited state decay was observed with an upper limit of 40 fs for the respective lifetimes. PMID- 19551203 TI - Effective temperature representation of quantum delocalization effects on the dynamics of NO(A(2)Sigma(+) <--X(2)Pi(+)) photoexcitation in Ne matrices. AB - The energetics of the optical excitation of the lowest Rydberg state of a nitric oxide molecule embedded in a neon matrix and the subsequent rearrangement of the solid host are investigated by classical molecular dynamics simulations. Quantum delocalization effects are incorporated through an effective temperature. The relevance of the inclusion of the anharmonicity of crystal site oscillations in the calculation of the effective temperature is evaluated. We show that representing zero point vibrations beyond the harmonic approximation improves the correspondence between the results of molecular dynamics simulations and pump probe experiments performed on this system. We explored both the steady state spectroscopy and the relaxation dynamics of the doped matrix in this improved representation. The analysis of the Fourier components of particle density reveals that the system simulated within the quantum thermal harmonic correction does not fulfill the translational symmetry of the host crystal. PMID- 19551204 TI - Stacked-structure-dependent photoelectrochemical properties of CdS nanoparticle/layered double hydroxide (LDH) nanosheet multilayer films prepared by layer-by-layer accumulation. AB - Inorganic multilayer films were prepared by layer-by-layer accumulation of positively charged layered double hydroxide (LDH) nanosheets and negatively charged CdS nanoparticles of different sizes. Nanoparticles were densely immobilized on LDH sheets to form a monolayer without coalescence into larger particles. The absorbance and photoluminescence intensity of immobilized CdS particles were enlarged with an increase in the accumulation number of the film. Hybrid films produced by accumulation of both monolayers of CdS particles (diameter: 5 nm) and those of smaller CdS particles (2.1 nm) exhibited characteristic photoluminescence spectra indicating the efficient energy transfer of photogenerated excitons from nanoparticle layers of smaller CdS particles to those of larger ones. LDH/CdS multilayers deposited on an F-doped SnO(2) (FTO) electrode behaved as an n-type semiconductor photoelectrode in an acetonitrile solution regardless of the size of the CdS particles immobilized, but their efficiency for photocurrent generation was greatly dependent on the stacked structure of the films. Accumulation of CdS particles of 2.1 nm in diameter on pre-coated LDH/CdS layers of 5 nm-sized CdS particles on FTO remarkably enhanced the photocurrent intensity in comparison to that in the case of accumulation of these two kinds of CdS particles in the opposite sequence. These observations can be explained by photoinduced electron transfer and energy transfer along with the band gap gradient in the films. PMID- 19551205 TI - Caging and solvent effects on the tautomeric equilibrium of 3-pyridone/3 hydroxypyridine in the ground state: a study in cyclodextrins and binary solvents. AB - The tautomeric equilibrium between 3-pyridone (3Py) and 3-hydroxypyridine (3HP) shows characteristic absorption peaks for the zwitterion form of 3Py in water that may be used as a probe of the hydrophobic nature inside macromolecules such as proteins and other biologically related systems. We studied this equilibrium in the ground state in aqueous cyclodextrins (CDs) and in binary solvent mixtures of 1,4-dioxane and water by absorption spectroscopy, and by ab initio calculations. Upon the addition of alpha-CD or beta-CD to an aqueous solution of the 3Py/3HP system, the absorbance intensity of the zwitterion tautomer decreases with a concomitant increase in the intensity of the enol tautomer of 3HP. The results reflect the nature of the tautomeric equilibrium and point to the hydrophobic environment inside the CD cavities. The effect of inclusion is noticeably less in the case of alpha-CD. This is attributed to the small cavity size of alpha-CD which sustains only partial inclusion. Upon the addition of gamma-CD, the intensity of the zwitterion tautomer slightly increased over that in water which is attributed to the direct interaction between the charged sides of the tautomer with the outer primary or secondary hydroxyls of the glycopyranose units of gamma-CD. This interaction is a result of the large cavity size of gamma-CD which does not support a stable complex. The largest caging effect was observed in 2,6-di-O-methyl-beta-CD (DMbeta-CD) which is an indication of a more hydrophobic environment around the guest. The large hydrophobicity of DMbeta-CD is due to the presence of the two methyl groups in the beta-CD derivative which reduce the amount of water inside the cavity upon encapsulation. In the binary mixtures of 1,4-dioxane and water, the change in the absorbance intensity of the enol and the zwitterion tautomers was analyzed quantitatively and three water molecules were found to solvate the polar centers of each tautomer. Ab initio calculations of the solvation of both tautomers by two and three water molecules were performed at the MP2/6-31++G(d,p) level. The calculations show that three water molecules are necessary to solvate the polar centers of each tautomer in a water network pattern. The results presented here suggest that the 3Py/3HP system represents a potentially useful new photophysical probe for supramolecular structures, particularly those involving inclusion. PMID- 19551206 TI - The photoionisation of two phenylcarbenes and their diazirine precursors investigated using synchrotron radiation. AB - Two phenylcarbenes, chlorophenylcarbene (CPC) and trifluoromethylphenylcarbene (TFPC), were generated by jet flash pyrolysis of diazirine precursors. Their photoionisation was studied by photoelectron-photoion coincidence spectroscopy using synchrotron radiation. For CPC we determined an adiabatic ionisation energy (IE) of 8.15 eV and a vertical IE of 9.3 eV. For TFPC we obtained an adiabatic IE of 8.47 eV and a vertical IE of 8.95 eV. The photoelectron spectra are broad and unstructured due to a large increase in the angle between the phenyl group, carbene centre and the substituent (Cl or CF(3)). The geometry change upon ionisation is more pronounced for CPC. Being a singlet arylcarbene, CPC is more strongly bent in the neutral ground state than the triplet TFPC. In addition, the bond between the carbenic centre and the chlorine atom shortens upon ionisation, because the radical cation is stabilised by the non-bonding electrons of the Cl through a mesomeric effect. The photoionisation and dissociative photoionisation of the diazirine precursors are also explored. The CPC precursor, 3-chloro-3 phenyldiazirine, undergoes complete dissociative photoionisation and only the CPC(+) fragment is observed above 8.8 eV. For 3-trifluoromethyl-3-phenyldiazirine some molecular ions can be observed above 9.05 eV. However, dissociative photoionisation also sets in right at the ionisation threshold. A fit to the data yields an appearance energy of AE (0 K) approximately 9.27 eV. PMID- 19551207 TI - Photolysis in aqueous aerosols: 300 nm yields of Fe2+ from a ferrioxalate actinometer and of OH radical from nitrate ions. AB - 300 nm photolysis yields of Fe(2+) from potassium ferrioxalate and of OH from nitrate ion have been measured in aqueous aerosols, the yield from ferrioxalate in a bulk solution being used to measure the light intensity. Mie theory was used to calculate effective cross-sections for absorption and scattering of light by the aerosol droplets. Yields of OH from nitrate ion have been measured with benzoic acid and carbon monoxide radical scavengers. The photolysis yield of Fe(2+) from ferrioxalate was found to be enhanced in the aerosol by a factor of 48 +/- 17. This enhancement is believed to be real, and is attributed to surfactant behaviour that results in the presence of a high concentration of ferrioxalate in a region of high light intensity near the droplet surface. The experiments with benzoic acid indicate that the yield of OH from nitrate in aerosol droplets is not significantly different from the yield in bulk solution. The CO experiments appear to indicate that the total OH production in the aerosol is enhanced over that in bulk solution by a factor of 10 +/- 3, but this number is not considered reliable. PMID- 19551208 TI - Tuned lifetime, at the ensemble and single molecule level, of a xanthenic fluorescent dye by means of a buffer-mediated excited-state proton exchange reaction. AB - The photophysical behaviour of the new fluorescein derivative 9-[1-(2-methyl-4 methoxyphenyl)]-6-hydroxy-3H-xanthen-3-one has been explored by using absorption, and steady-state, time-resolved and single-molecule fluorescence measurements. The apparent ground-state acidity constant of the dye determined by both the absorbance and steady-state fluorescence is almost independent of the added buffer and salt concentrations. The excited-state proton exchange reaction around the physiological pH becomes reversible upon addition of phosphate buffer, inducing a pH-dependent change of the steady-state fluorescence and decay times. Fluorescence decay traces, collected as a function of total buffer concentration and pH, were analyzed by global compartmental analysis (GCA) to elucidate the values of the excited-state rate constants. The features of this system make the fluorescence decays monoexponential at pH values and phosphate buffer concentrations higher than 6.10 and 0.2 M respectively, with the possibility of tuning the fluorescence lifetime value by changing pH or buffer concentrations. The tuned lifetimes obtained by means of phosphate concentration at constant pH have also been recovered at the single-molecule level. PMID- 19551210 TI - Surface reconstruction of Pt nanocrystals interacting with gas atmosphere. Bridging the pressure gap with in situ diffraction. AB - A carefully designed in situ XRD experiment when guided by atomistic simulations can provide data on the atomistic structure of a surface layer of platinum nanoclusters. Even the adsorption process for a strongly bonded adsorbate can be monitored and interpreted, providing data that are not available from other techniques. The data reported here present the first observation of surface reconstruction of nanocrystals by X-ray diffraction known to the authors. We were able to observe repeatable in situ evolution of Pt nanocrystal diffraction peak positions on exchange of gas atmosphere from hydrogen to helium. Experiments at room temperature and at 373 K shows various hydrogen desorption rate in He atmosphere but a very similar rate of an average lattice constant change with clearly separated desorption and reconstruction phases. The hydrogen desorption rate has been shown to be controlled by a slower process-hydrogen spillover and its activation energy was estimated. Diffraction peaks of Pt on exposition to O(2) shift at various degree to lower angles due to surface relaxation-the effect being particle-size-dependent and illustrating elongation of surface Pt-Pt bonds caused by adsorption. The results show the possibility for XRD to become a nanosurface science tool enabling the combination of structure analysis with adsorption/desorption measurements within the pressure gap and material gap. PMID- 19551209 TI - Photophysics of manisyl-substituted 2-pyridin-2-yl-1,10-phenanthrolines. Dual emission dependent on structure and solvent. AB - Systematic variation of the substitution pattern of 2-pyridin-2-yl-1,10 phenanthrolines with 4-methoxy-2,6-dimethylphenyl (manisyl) groups alpha, beta, and gamma to the nitrogen atoms results in a 3 x 3 array of pyridyl phenanthrolines displaying low to high quantum yields (Phi(f) = 0.03-0.60). Photophysical studies elucidated a duality of emissive states: a weakly emissive, locally excited state, similar to the (1)pi,pi state of phenanthroline; and a strongly emissive, charge-transfer state, dependent on manisyl regiochemistry and solvent polarity. Ab initio calculations underscore the similarities in the electronic structures of phenanthroline and pyridyl-phenanthrolines rather than between terpyridine and pyridyl-phenanthroline. PMID- 19551211 TI - Electrochemical reactions at a porphyrin-copper interface. AB - The structure and reactivity of a Cu(100) single crystal electrode surface covered with free base meso-tetra (N-methyl-4-pyridinium) porphyrin (abbreviated as H(2)TMPyP) as a function of electrode potential have been investigated with cyclic voltammetry (CV), electrochemical scanning tunneling microscopy (ECSTM), and UV-Vis and Raman spectroscopy. The well-ordered self-assembled layer of the porphyrin is consistent with the adsorption of the reduced porphyrin species after the first two-electron reduction step. The copper dissolution reaction in the presence of the stable self-assembled porphyrin layer starts at step edges on both upper and lower terraces and coincides with the preferential oxidation of reduced porphyrin species at step sites. The dissolved copper cations are incorporated into the free base porphyrin molecules leading to the formation of CuTMPyP. As a consequence this new species accumulates in the solution with time and a copper redeposition in the cathodic potential scan is lacking. PMID- 19551212 TI - An embedded cluster study of the formation of water on interstellar dust grains. AB - The formation of water in the interstellar medium from hydrogen and oxygen atoms on a pristine olivine surface (forsterite (010)) is investigated with an embedded cluster approach. The 55-atom quantum cluster is described at the density functional level while the remaining 1629 atoms of the surface cluster are described with atomistic potentials. Transition states are most easily calculated with our modified implementation of the climbing-image nudged elastic band method in ChemShell. With these computational techniques, we find that gas phase hydrogen atoms can chemisorb (-102 kJ mol(-1)) without an activation barrier on the forsterite (010) surface, concomitantly creating a surface electron at the adjacent magnesium atom site. Subsequently, an oxygen atom chemisorbs strongly to this surface electron site (-432 kJ mol(-1)). The rearrangement of the adjacently chemisorbed O and H to a chemisorbed OH-radical is endothermic by 4 kJ mol(-1) and activated by 27 kJ mol(-1). This chemisorbed OH can then react barrierlessly with a second hydrogen atom to yield adsorbed water (-511 kJ mol(-1)). Alternatively, if O and H do not recombine to form OH, but instead thermally equilibrate, a second hydrogen atom can react with the chemisorbed oxygen atom ( 501 kJ mol(-1)) to yield dissociatively adsorbed water (OH(-) and H(+)), which then can rearrange to associatively adsorbed water (-5 kJ mol(-1), DeltaE(double dagger) = 18 kJ mol(-1)) or gas phase water (+91 kJ mol(-1)). The formation of water on a bare dust grain from hydrogen and oxygen atoms is thus catalysed by an olivine surface by stabilising the reaction intermediates and product. Since the reaction proceeds via three chemisorbed intermediates, thermal equilibration is facilitated and back-dissociation of the freshly formed reaction products OH and H(2)O would not occur as frequently as it would in the gas phase or when the reactants are physisorbed on a surface. PMID- 19551213 TI - Time-resolved spectroscopy of dye-labeled photoactive yellow protein suggests a pathway of light-induced structural changes in the N-terminal cap. AB - The photoreceptor PYP responds to light activation with global conformational changes. These changes are mainly located in the N-terminal cap of the protein, which is approximately 20 A away from the chromophore binding pocket and separated from it by the central beta-sheet. The question of the propagation of the structural change across the central beta-sheet is of general interest for the superfamily of PAS domain proteins, for which PYP is the structural prototype. Here we measured the kinetics of the structural changes in the N terminal cap by transient absorption spectroscopy on the ns to second timescale. For this purpose the cysteine mutants A5C and N13C were prepared and labeled with thiol reactive 5-iodoacetamidofluorescein (IAF). A5 is located close to the N terminus, while N13 is part of helix alpha1 near the functionally important salt bridge E12-K110 between the N-terminal cap and the central anti-parallel beta sheet. The absorption spectrum of the dye is sensitive to its environment, and serves as a sensor for conformational changes near the labeling site. In both labeled mutants light activation results in a transient red-shift of the fluorescein absorption spectrum. To correlate the conformational changes with the photocycle intermediates of the protein, we compared the kinetics of the transient absorption signal of the dye with that of the p-hydroxycinnamoyl chromophore. While the structural change near A5 is synchronized with the rise of the I(2) intermediate, which is formed in approximately 200 mus, the change near N13 is delayed and rises with the next intermediate I(2)', which forms in approximately 2 ms. This indicates that different parts of the N-terminal cap respond to light activation with different kinetics. For the signaling pathway of photoactive yellow protein we propose a model in which the structural signal propagates from the chromophore binding pocket across the central beta-sheet via the N-terminal region to helix alpha1, resulting in a large change in the protein conformation. PMID- 19551214 TI - Formation of gold and gold sulfide nanoparticles and mesoscale intermediate structures in the reactions of aqueous HAuCl4 with sulfide and citrate ions. AB - The effects of the molar ratio of sodium sulfide to chloroauric acid in the range of 0.5 to 5 and the time factor on the formation of the nanoparticles (NPs) of metallic Au, Au(2)S or their mixtures have been studied applying in situ and ex situ techniques (UV-Vis absorption spectroscopy, potentiometry, TEM, SPM, SERS, XPS). The products and intermediates have been compared with those for the reduction of chloroaurate with citrate ions and combinations of citrate and sulfide ions. An increase in the concentration of sulfide ions accelerates the reduction of Au(iii) complexes but hinders the nucleation and growth of Au NPs, resulting in a prolonged period before the appearance of plasmon peaks. The electrochemical potential is not directly associated with the plasmon intensities, although the potential sharply decreases simultaneously with a blue shift of the near-IR peak emerging with the Na(2)S/HAuCl(4) ratios of 0.5 to 1.5. It was concluded that the peak is due to longitudinal plasmon resonance of gold nanoplates. Au(2)S NPs, the nucleation of which is effectively inhibited, and probably some structures and fragments visible in TEM and AFM, including 2-5 nm Au NPs, crystallize in part outside the solutions. The evidence of partially liquid mesoscale structures comprising intermediate gold species as precursors of nanoparticles is presented, and their origin, ex situ transformation and role in the reaction mechanisms are discussed. PMID- 19551216 TI - Solubilization of flurbiprofen within non-ionic Tween 20 surfactant micelles: a 19F and 1H NMR study. AB - The solubilization of the poorly water soluble anti-inflammatory drug flurbiprofen in non-ionic Tween 20 surfactant micellar solutions was studied by both (19)F and (1)H NMR spectroscopy in an acidic environment. These non destructive techniques allowed us to investigate the effect of temperature cycling in situ. Using (19)F NMR, an increased solubilisation capacity was observed as the temperature increased. This effect became more pronounced above the cloud point, which was reduced by more than 30 degrees C in the presence of an excess of flurbiprofen. Upon clouding, peak splitting was observed in the (19)F spectrum, which indicates that two pools of solubilised flurbiprofen exist that are in slow exchange on the NMR frequency timescale. The clouding and solubilization processes were found to be reversible, albeit with slow kinetics. Based on chemical shift differences of both Tween 20 and flurbiprofen, as well as NOESY experiments, the flurbiprofen was found to be accumulated within the palisade layer of the Tween 20 micelles. PMID- 19551215 TI - Morphology and optical responses of SERS active pi-conjugated poly(N-ethyl-2 ethynylpyridinium iodide)/Ag nanocomposite systems. AB - The influence of the poly(N-ethyl-2-ethynylpyridinium iodide) (PEEP-I) concentration on the morphology and optical properties of nanocomposite systems prepared by mixing the polymer solution with a hydrosol of ca. 9 nm Ag nanoparticles (NPs) was investigated by a combination of surface plasmon extinction (SPE) measurements, transmission electron microscope (TEM) imaging and surface-enhanced Raman spectroscopy (SERS). The PEEP-I concentration was found to have a strong impact on the assembly of Ag NPs and, consequently, on the optical responses of the composite systems. At low polymer concentrations in the composite (corresponding to ca. 50-1800 monomer units/NP), the formation of fractal aggregates was observed. In particular, the average fractal dimension D = 1.9 +/- 0.1 was determined for aggregates in the system with 5 x 10(-6) M polymer concentration. By contrast, in systems with polymer concentrations higher than about 1 x 10(-5) M, relatively small aggregates of Ag NPs with large interparticle distances were formed. The differences in the morphology of the composite systems with various polymer concentrations manifested themselves clearly in their SPE spectra. Furthermore, upon optical excitation with appropriate wavelengths (488.0 and 514.5 nm), the fractal aggregates acted as carriers of "hot spots", i.e. strong, localized, nanoscale optical fields, from which intense and well resolved SERS spectra of the polymer were obtained. PMID- 19551217 TI - New insight into the nanostructure of ionic liquids: a small angle X-ray scattering (SAXS) study on liquid tri-alkyl-methyl-ammonium bis(trifluoromethanesulfonyl)amides and their mixtures. AB - We report a small angle X-ray scattering study on the liquid phase of a series of room temperature ionic liquids and their binary mixtures. The ionic liquids studied belong to the tri-alkyl-methyl-ammonium family with bis(trifluoromethanesulfonyl)amide as the anion and were studied as a function of alkyl chain length. These ionic liquids were found to exhibit marked nanoscale ordering in their isotropic liquid state as judged from the small angle X-ray scattering. The observed structural ordering is of supramolecular order and depends strongly on the length of the cation hydrophobic side chain. Moreover, the data can be analyzed on the basis of a disordered smectic A phase, consisting of strongly interdigitated bilayers that sequester the ionic liquid into polar and hydrophobic regions. These findings were also found to be consistent with density data of these molten salts. Additionally, we demonstrate that this experimentally observed nanostructuring can further be fine-tuned using binary mixtures. PMID- 19551219 TI - Non-perturbative magnetic phenomena in closed-shell paramagnetic molecules. AB - By means of non-perturbative ab initio calculations, it is shown that paramagnetic closed-shell molecules are characterized by a strongly non-linear magnetic response, whose main feature consists of a paramagnetic-to-diamagnetic transition in a strong magnetic field. The physical origin of this phenomenon is rationalised on the basis of an analytical model based on molecular orbital theory. For the largest molecules considered here, the acepleiadylene dianion and the corannulene dianion, the transition field is of the order of 10(3) T, about one order of magnitude larger than the magnetic field strength currently achievable in experimental settings. However, our simple model suggests that the paramagnetic-to-diamagnetic transition is a universal property of paramagnetic closed-shell systems in strong magnetic fields, provided no singlet-triplet level crossing occurs for fields smaller than the critical transition field. Accordingly, fields weaker than 100 T should suffice to trigger the predicted transition for systems whose size is still well within the (medium-large) molecular domain, such as hypothetical antiaromatic rings with less than one hundred carbon atoms. PMID- 19551218 TI - In situ XAS and IR studies on Cu:SAPO-5 and Cu:SAPO-11: the contributory role of monomeric linear copper(i) species in the selective catalytic reduction of NOx by propene. AB - Cu:SAPO-5 and Cu:SAPO-11 were prepared by conventional and hydrothermal ion exchange. Copper incorporation is increased six-fold by hydrothermal ion exchange relative to conventional methods. In all cases, the amount of copper taken up by SAPO-11 is superior to uptake in SAPO-5. Copper is divalent and in tetragonally distorted octahedral environments in the as-prepared samples independent of the method of incorporation for both systems. The local structures about the metal and the valence states associated with the different steps in the selective catalytic reduction of NO(x) in the presence of propene (SCR-HC) have been investigated using X-ray absorption spectroscopy (XAS). For both the Cu:SAPO-5 and Cu:SAPO-11 systems, heating in helium partially autoreduces copper(ii) to copper(i). Following activation in oxygen, propene causes further reduction to copper(i) in all four samples as shown by the evolution of an intense pre-edge diagnostic feature. XANES analysis reveal this to be characteristic of monomeric linear two coordinate copper(i) species. This is a prime example of a pre-edge peak with such a high intensity being observed in the solid state. This is supported by IR where peaks attributed to bidentate copper were observed for Cu:SAPO-11/HT. For all four samples NO(x) partially reoxidises the copper(i) formed in the helium and propene steps. Ion exchanged Cu:SAPO-5 and Cu:SAPO-11 exhibit low activity in reducing NO(x) by propene in an oxygen rich environment. The role of the copper ion during NO adsorption was studied using in situ infra red spectroscopy. The activity of copper exchanged materials is governed by both the degree of reducibility of copper(ii) and the ease of reversing the valence states with the structural characteristics of the parent materials playing a crucial role. PMID- 19551220 TI - Time-domain electrostatic force spectroscopy on nanostructured lithium-ion conducting glass ceramics: analysis and interpretation of relaxation times. AB - The nanoscopic electrical properties of LiAlSiO(4) glass ceramics with different degrees of crystallinity chi were studied by means of time-domain electrostatic force spectroscopy (TDEFS). Thereby, a faster relaxation process due to lithium ion movements in the glassy phase and a slower process due to lithium ion movements in the crystallites could be distinguished. Over a broad range of crystallinity values, the TDEFS relaxation times of both processes are Arrhenius activated, with an activation energy being essentially independent of chi and with a pre-exponential factor depending in a systematic fashion on chi. With increasing crystallinity, the pre-exponential factor of the faster process (glassy phase) increases, while that of the slower process (crystallites) decreases. In order to explain this observation, we consider simple equivalent circuit models for the capacitance relaxation of the system AFM tip/gap/sample. PMID- 19551221 TI - Defect interactions and ionic transport in scandia stabilized zirconia. AB - Classical molecular dynamics simulation has been used to study ionic transport in scandia-stabilized zirconia, as well as scandia and yttria-co-doped zirconia, as a function of temperature and composition. The oxygen diffusion coefficient shows a peak at a composition of 6 mol% Sc(2)O(3). At 1125 K and higher temperatures, oxygen vacancies prefer to be second nearest neighbours to yttrium ions and first neighbours to scandium ions, because the defect interactions in scandia stabilized zirconia are governed mainly by electrostatic effects. Oxygen migration between cation tetrahedra is impeded less effectively by Sc-Sc edges than by Y-Y edges. The formation of neutral dopant-anion vacancy clusters is favoured, in agreement with recent nuclear magnetic resonance observations. PMID- 19551222 TI - Crystal structure prediction for cyclotrimethylene trinitramine (RDX) from first principles. AB - Crystal structure prediction and molecular dynamics methods were applied to the cyclotrimethylene trinitramine (RDX) crystal to explore the stability rankings of various polymorphs using a recently developed nonempirical potential energy function that describes the RDX dimer interactions. The energies of 500 high density structures resulting from molecular packing were minimized and the 14 lowest-energy structures were subjected to isothermal-isostress molecular dynamics (NsT-MD) simulations. For both crystal structure prediction methods and molecular dynamics simulations, the lowest-energy polymorph corresponded to the experimental structure; furthermore, the lattice energy of this polymorph was lower than that of the other polymorphs by at least 1.1 kcal mol(-1). Crystal parameters and densities of the low-energy crystal produced by the NsT-MD simulations matched those of the experimental crystal to within 1% of density and cell edge lengths and 0.01 degrees of the cell angle. The arrangement of the molecules within the time-averaged unit cell were in equally outstanding agreement with experiment, with the largest deviation of the location of the molecular mass centers being less than 0.07 A and the largest deviation in molecular orientation being less than 2.8 degrees . NsT-MD simulations were also used to calculate crystallographic parameters as functions of temperature and pressure and the results were in a reasonable agreement with experiment. PMID- 19551225 TI - Injuries: the neglected burden in developing countries. PMID- 19551226 TI - Why health communication is important in public health. PMID- 19551227 TI - Labs form a new front against deadly pathogens. PMID- 19551228 TI - WHO takes lead on health as UN tackles crises. PMID- 19551229 TI - Greater needs, limited access. PMID- 19551230 TI - Mobilizing the honest majority to fight health-sector fraud. PMID- 19551233 TI - The lifetime risk of maternal mortality: concept and measurement. AB - OBJECTIVE: The lifetime risk of maternal mortality, which describes the cumulative loss of life due to maternal deaths over the female life course, is an important summary measure of population health. However, despite its interpretive appeal, the lifetime risk of dying from maternal causes can be defined and calculated in various ways. A clear and concise discussion of both its underlying concept and methods of measurement is badly needed. METHODS: I define and compare a variety of procedures for calculating the lifetime risk of maternal mortality. I use detailed survey data from Bangladesh in 2001 to illustrate these calculations and compare the properties of the various risk measures. Using official UN estimates of maternal mortality for 2005, I document the differences in lifetime risk derived with the various measures. FINDINGS: Taking sub-Saharan Africa as an example, the range of estimates for the 2005 lifetime risk extends from 3.41% to 5.76%, or from 1 in 29 to 1 in 17. The highest value resulted from the method used for producing official UN estimates for the year 2000. The measure recommended here has an intermediate value of 4.47%, or 1 in 22. CONCLUSION: There are strong reasons to consider the calculation method proposed here more accurate and appropriate than earlier procedures. Accordingly, it was adopted for use in producing the 2005 UN estimates of the lifetime risk of maternal mortality. By comparison, the method used for the 2000 UN estimates appears to overestimate this important measure of population health by around 20%. PMID- 19551234 TI - Emergency triage assessment for hypoxaemia in neonates and young children in a Kenyan hospital: an observational study. AB - OBJECTIVE: To describe the prevalence of hypoxaemia in children admitted to a hospital in Kenya for the purpose of identifying clinical signs of hypoxaemia for emergency triage assessment, and to test the hypothesis that such signs lead to correct identification of hypoxaemia in children, irrespective of their diagnosis. METHODS: From 2002 to 2005 we prospectively collected clinical data and pulse oximetry measurements for all paediatric admissions to Kilifi District Hospital, Kenya, irrespective of diagnosis, and assessed the prevalence of hypoxaemia in relation to the WHO clinical syndromes of 'pneumonia' on admission and the final diagnoses made at discharge. We used the data collected over the first three years to derive signs predictive of hypoxaemia, and data from the fourth year to validate those signs. FINDINGS: Hypoxemia was found in 977 of 15 289 (6.4%) of all admissions (5% to 19% depending on age group) and was strongly associated with inpatient mortality (age-adjusted risk ratio: 4.5; 95% confidence interval, CI: 3.8-5.3). Although most hypoxaemic children aged > 60 days met the WHO criteria for a syndrome of 'pneumonia' on admission, only 215 of the 693 (31%) such children had a final diagnosis of lower respiratory tract infection (LRTI). The most predictive signs for hypoxaemia included shock, a heart rate < 80 beats per minute, irregular breathing, a respiratory rate > 60 breaths per minute and impaired consciousness. However, 5-15% of the children who had hypoxaemia on admission were missed, and 18% of the children were incorrectly identified as hypoxaemic. CONCLUSION: The syndromes of pneumonia make it possible to identify most hypoxaemic children, including those without LRTI. Shock, bradycardia and irregular breathing are important predictive signs, and severe malaria with respiratory distress is a common cause of hypoxaemia. Overall, however, clinical signs are poor predictors of hypoxaemia, and using pulse oximetry in resource-poor health facilities to target oxygen therapy is likely to save costs. PMID- 19551236 TI - Financial barriers to HIV treatment in Yaounde, Cameroon: first results of a national cross-sectional survey. AB - OBJECTIVE: To assess the extent to which user fees for antiretroviral therapy (ART) represent a financial barrier to access to ART among HIV-positive patients in Yaounde, Cameroon. METHODS: Sociodemographic, economic and clinical data were collected from a random sample of 707 HIV-positive patients followed up in six public hospitals of the capital city (Yaounde) and its surroundings through face to-face interviews carried out by trained interviewers independently from medical staff and medical questionnaires filled out by prescribing physicians. Logistic regression models were used to identify factors associated with self-reported financial difficulties in purchasing ART during the previous 3 months. FINDINGS: Of the 532 patients treated with ART at the time of the survey, 20% reported financial difficulty in purchasing their antiretroviral drugs during the previous 3 months. After adjustment for socioeconomic and clinical factors, reports of financial difficulties were significantly associated with lower adherence to ART (odds ratio, OR: 0.24; 95% confidence interval, CI: 0.15-0.40; P < 0.0001) and with lower CD4+ lymphocyte (CD4) counts after 6 months of treatment (OR: 2.14; 95% CI: 1.15-3.96 for CD4 counts < 200 cells/microl; P = 0.04). CONCLUSION: Removing a financial barrier to treatment with ART by eliminating user fees at the point of care delivery, as recommended by WHO, could lead to increased adherence to ART and to improved clinical results. New health financing mechanisms based on the public resources of national governments and international donors are needed to attain universal access to drugs and treatment for HIV infection. PMID- 19551235 TI - How does satisfaction with the health-care system relate to patient experience? AB - OBJECTIVE: To explore what determines people's satisfaction with the health-care system above and beyond their experience as patients. METHODS: Data on health system responsiveness, which refers to the manner and environment in which people are treated when they seek health care, provides a unique opportunity to better understand the determinants of people's satisfaction with the health-care system and how strongly this is influenced by an individual's experience as a patient. The data were obtained from 21 European Union countries in the World Health Survey for 2003. Additive ordinary least-squares regression models were used to assess the extent to which variables commonly associated with satisfaction with the health-care system, as recorded in the literature, explain the variation around the concept of satisfaction. A residual analysis was used to identify other predictors of satisfaction with the health-care system. FINDINGS: Patient experience was significantly associated with satisfaction with the health-care system and explained 10.4% of the variation around the concept of satisfaction. Other factors such as patient expectations, health status, type of care, and immunization coverage were also significant predictors of health system satisfaction; although together they explained only 17.5% of the observed variation, while broader societal factors may largely account for the unexplained portion of satisfaction with the health-care system. CONCLUSION: Contrary to published reports, people's satisfaction with the health-care system depends more on factors external to the health system than on the experience of care as a patient. Thus, measuring the latter may be of limited use as a basis for quality improvement and health system reform. PMID- 19551237 TI - Estimating pregnancy-related mortality from census data: experience in Latin America. AB - OBJECTIVE: To assess the feasibility of measuring maternal mortality in countries lacking accurate birth and death registration through national population censuses by a detailed evaluation of such data for three Latin American countries. METHODS: We used established demographic techniques, including the general growth balance method, to evaluate the completeness and coverage of the household death data obtained through population censuses. We also compared parity to cumulative fertility data to evaluate the coverage of recent household births. After evaluating the data and adjusting it as necessary, we calculated pregnancy-related mortality ratios (PRMRs) per 100 000 live births and used them to estimate maternal mortality. FINDINGS: The PRMRs for Honduras (2001), Nicaragua (2005) and Paraguay (2002) were 168, 95 and 178 per 100 000 live births, respectively. Surprisingly, evaluation of the data for Nicaragua and Paraguay showed overreporting of adult deaths, so a downward adjustment of 20% to 30% was required. In Honduras, the number of adult female deaths required substantial upward adjustment. The number of live births needed minimal adjustment. The adjusted PRMR estimates are broadly consistent with existing estimates of maternal mortality from various data sources, though the comparison varies by source. CONCLUSION: Census data can be used to measure pregnancy related mortality as a proxy for maternal mortality in countries with poor death registration. However, because our data were obtained from countries with reasonably good statistical systems and literate populations, we cannot be certain the methods employed in the study will be equally useful in more challenging environments. Our data evaluation and adjustment methods worked, but with considerable uncertainty. Ways of quantifying this uncertainty are needed. PMID- 19551238 TI - The persistence of tuberculosis in the age of DOTS: reassessing the effect of case detection. AB - OBJECTIVE: To investigate whether short-term annual declines of 5-10% in the incidence of tuberculosis (TB) can be sustained over the long term by maintaining high case detection rates (CDRs). METHODS: We constructed a compartmental difference-equation model of a TB epidemic in a hypothetical population of constant size with a treatment success rate of 85%. The impact of CDR on TB incidence was then investigated by generating an equilibrium population with no TB case detection and increasing the smear-positive CDR under two scenarios: (i) rapid expansion by 10% per year to a CDR of 80% after 8 years, and (ii) gradual expansion by 1% per year to a CDR of 90% after 90 years. The model was applied in two hypothetical populations: one without HIV and the other with a stable HIV incidence representative of the African Region. The CDR for smear-negative TB was assumed to be a constant fraction of the smear-positive CDR. FINDINGS: In the absence of a TB control programme, the projected annual incidence of TB was 513 cases per 100 000 population, with a point prevalence of 1233 per 100 000 and an annual TB-specific mortality rate of 182 per 100 000. Immediately increasing the TB CDR from 0% to 70% caused a 74% reduction in TB incidence within 10 years. However, once case detection stabilized at any constant level < 80%, projected TB incidence also stabilized. Ten years after a CDR of 70% was reached, the annual decline in TB incidence was < 1.5%, regardless of how rapidly case detection was scaled up and despite wide variation of all model parameters. CONCLUSION: While improved CDRs have a dramatic short-term effect on TB incidence, maintaining those rates, even at current target levels, may not reduce long-term incidence by more than 1-2% per year. TB control programmes and researchers should vigorously pursue improvements in case detection, regardless of current CDRs. PMID- 19551239 TI - Population-based simulations of influenza pandemics: validity and significance for public health policy. AB - OBJECTIVE: To examine the validity and usefulness of pandemic simulations aimed at informing practical decision-making in public health. METHODS: We recruited a multidisciplinary group of nine experts to assess a case-study simulation of influenza transmission in a Swedish county. We used a non-statistical nominal group technique to generate evaluations of the plausibility, formal validity (verification) and predictive validity of the simulation. A health-effect assessment structure was used as a framework for data collection. FINDINGS: The unpredictability of social order during disasters was not adequately addressed by simulation methods; even minor disruptions of the social order may invalidate key infrastructural assumptions underpinning current pandemic simulation models. Further, a direct relationship between model flexibility and computation time was noted. Consequently, simulation methods cannot, in practice, support integrated modifications of microbiological, epidemiological and spatial submodels or handle multiple parallel scenarios. CONCLUSION: The combination of incomplete surveillance data and simulation methods that neglect social dynamics limits the ability of national public health agencies to provide policy-makers and the general public with the critical and timely information needed during a pandemic. PMID- 19551241 TI - The rationale for integrated childhood meningoencephalitis surveillance: a case study from Cambodia. AB - PROBLEM: Recent progress in vaccine availability and affordability has raised prospects for reducing death and disability from neurological infections in children. In many Asian countries, however, the epidemiology and public health burden of neurological diseases such as Japanese encephalitis and bacterial meningitis are poorly understood. APPROACH: A sentinel surveillance system for Japanese encephalitis was developed and embedded within the routine meningoencephalitis syndromic surveillance system in Cambodia in 2006. The sentinel surveillance system was designed so surveillance and laboratory testing for other etiologies of neurological infection could be incorporated. LOCAL SETTING: The Communicable Disease Control department of the Ministry of Health in Cambodia worked with partners to establish the sentinel surveillance system. RELEVANT CHANGES: The sentinel surveillance system has provided important information on the disease burden of Japanese encephalitis in Cambodia and is now providing a platform for expansion to incorporate laboratory testing for other vaccine-preventable neurological infections in children. LESSONS LEARNED: Sentinel surveillance systems, when linked to syndromic reporting systems, can characterize the epidemiology of meningoencephalitis and identify the proportion of hospital-based neurological infection in children that is vaccine preventable. Integrated systems enable consistency in data collection, analysis and information dissemination, and they enhance the capacity of public health managers to provide more credible and integrated information to policy-makers. This will assist decision-making about the potential role of immunization in reducing the incidence of childhood neurological infections. PMID- 19551240 TI - Promoting the health of marginalized populations in Ecuador through international collaboration and educational innovations. AB - This paper examines two innovative educational initiatives for the Ecuadorian public health workforce: a Canadian-funded Masters programme in ecosystem approaches to health that focuses on building capacity to manage environmental health risks sustainably; and the training of Ecuadorians at the Latin American School of Medicine in Cuba (known as Escuela Latinoamericana de Medicina in Spanish). We apply a typology for analysing how training programmes address the needs of marginalized populations and build capacity for addressing health determinants. We highlight some ways we can learn from such training programmes with particular regard to lessons, barriers and opportunities for their sustainability at the local, national and international levels and for pursuing similar initiatives in other countries and contexts. We conclude that educational efforts focused on the challenges of marginalization and the determinants of health require explicit attention not only to the knowledge, attitudes and skills of graduates but also on effectively engaging the health settings and systems that will reinforce the establishment and retention of capacity in low- and middle-income settings where this is most needed. PMID- 19551242 TI - Rehabilitation of the injured child. PMID- 19551244 TI - Child injuries and violence: responding to a global challenge. PMID- 19551245 TI - Prevention is better than cure. PMID- 19551246 TI - Healing child soldiers. PMID- 19551247 TI - Fragile brain, handle with care. PMID- 19551248 TI - Child maltreatment comes out of the shadows. PMID- 19551249 TI - Education and legislation are key to preventing child injuries. PMID- 19551251 TI - Severe physical punishment: risk of mental health problems for poor urban children in Brazil. AB - OBJECTIVE: To examine the relationship between specific types of child mental health problems and severe physical punishment, in combination with other important known risk factors. METHODS: We conducted a cross-sectional study in Embu, Sao Paulo, Brazil, as the Brazilian component of a multicountry survey on abuse in the family environment. From a probabilistic sample of clusters that included all eligible households (women aged 15-49 years with a son or daughter < 18 years of age), we randomly selected one mother-child pair per household (n = 813; attrition rate: 17.6%). This study focused on children aged 6-17 years (n = 480). Child Behaviour Checklist CBCL/6-18 was used to identify children with internalizing problems only, externalizing problems only, and both internalizing and externalizing problems (comorbidity). Severe physical punishment was defined as being hit with an object, being kicked, choked, smothered, burnt, scalded, branded, beaten or threatened with a weapon. We examined other potential correlates from four domains: child (gender, age, ever witnessing marital violence); mother (education, unemployment, anxiety or depression, marital violence); father (absence, drunkenness); and family (socioeconomic status). The WHO Self-Reporting Questionnaire (SRQ-20) was used to identify maternal anxiety or depression (score > 7). Backward logistic regression analysis identified independent correlates and significant interactions. FINDINGS: Multivariate modelling showed that severe punishment was an independent correlate of comorbid internalizing and externalizing problems but was not associated with internalizing problems only. It increased the risk of externalizing problems alone only for children and adolescents not exposed to maternal anxiety or depression. Maternal anxiety or depression increased the risk only for children or adolescents not exposed to severe punishment. CONCLUSION: Severe punishment may be related to child mental health problems, with the mechanism depending on the type of problem. Its influence persists in the presence of family stressors such as the father's absence and maternal anxiety or depression. PMID- 19551253 TI - Child maltreatment prevention: a systematic review of reviews. AB - OBJECTIVE: To synthesize recent evidence from systematic and comprehensive reviews on the effectiveness of universal and selective child maltreatment prevention interventions, evaluate the methodological quality of the reviews and outcome evaluation studies they are based on, and map the geographical distribution of the evidence. METHODS: A systematic review of reviews was conducted. The quality of the systematic reviews was evaluated with a tool for the assessment of multiple systematic reviews (AMSTAR), and the quality of the outcome evaluations was assessed using indicators of internal validity and of the construct validity of outcome measures. FINDINGS: The review focused on seven main types of interventions: home visiting, parent education, child sex abuse prevention, abusive head trauma prevention, multi-component interventions, media based interventions, and support and mutual aid groups. Four of the seven - home visiting, parent education, abusive head trauma prevention and multi-component interventions - show promise in preventing actual child maltreatment. Three of them - home visiting, parent education and child sexual abuse prevention - appear effective in reducing risk factors for child maltreatment, although these conclusions are tentative due to the methodological shortcomings of the reviews and outcome evaluation studies they draw on. An analysis of the geographical distribution of the evidence shows that outcome evaluations of child maltreatment prevention interventions are exceedingly rare in low- and middle-income countries and make up only 0.6% of the total evidence base. CONCLUSION: Evidence for the effectiveness of four of the seven main types of interventions for preventing child maltreatment is promising, although it is weakened by methodological problems and paucity of outcome evaluations from low- and middle-income countries. PMID- 19551254 TI - Violence-related injury of children in Israel: age-dependent pattern. AB - OBJECTIVE: To characterize the population of children hospitalized as a result of violence. METHODS: This retrospective study used data from the Israeli National Trauma Registry on patients aged 0-17 years hospitalized for trauma during 1998 2006. Of 65,430 patients, 2060 (3.1%) had violence-related injuries. Descriptive statistics and bivariate analysis were used to characterize injury patterns and multivariate analysis was used to identify factors associated with severe injury. FINDINGS: Half the victims of violence < 18 years of age were 15-17 years old. Most were boys. Violence-related trauma occurred more often than other trauma in the street, at school or in a public place or leisure facility, but less often at home. Unarmed brawling and stabbing were the most common types of violence. Brawling affected Jewish children more, while non-Jewish children were more often injured by firearms. The boundaries of age groups with different injury rates corresponded to the 'institutional' childhood stages of the Israeli educational system: the violence-related injury rate dropped after the first year of life, stayed low during kindergarten, rose slightly during elementary school and rose steeply during secondary and high school. The percentage of males increased with each age group. The street became more dangerous with age, while school and home became safer. Adolescents aged 15-17 years, newborn infants and those injured by firearms had the highest risk of severe injury. CONCLUSION: Age and type of violence were the most important predictors of violence frequency and severity. Ethnicity lost importance when adjusted by these factors. Further research on their influence on violence-related injury is needed. PMID- 19551252 TI - Global childhood unintentional injury surveillance in four cities in developing countries: a pilot study. AB - OBJECTIVE: To determine the frequency and nature of childhood injuries and to explore the risk factors for such injuries in low-income countries by using emergency department (ED) surveillance data. METHODS: This pilot study represents the initial phase of a multi-country global childhood unintentional injury surveillance (GCUIS) project and was based on a sequential sample of children < 11 years of age of either gender who presented to selected EDs in Bangladesh, Colombia, Egypt and Pakistan over a 3-4 month period, which varied for each site, in 2007. FINDINGS: Of 1559 injured children across all sites, 1010 (65%) were male; 941 (60%) were aged >or= 5 years, 32 (2%) were < 1 year old. Injuries were especially frequent (34%) during the morning hours. They occurred in and around the home in 56% of the cases, outside while children played in 63% and during trips in 11%. Of all the injuries observed, 913 (56%) involved falls; 350 (22%), road traffic injuries; 210 (13%), burns; 66 (4%), poisoning; and 20 (1%), near drowning or drowning. Falls occurred most often from stairs or ladders; road traffic injuries most often involved pedestrians; the majority of burns were from hot liquids; poisonings typically involved medicines, and most drowning occurred in the home. The mean injury severity score was highest for near drowning or drowning (11), followed closely by road traffic injuries (10). There were 6 deaths, of which 2 resulted from drowning, 2 from falls and 2 from road traffic injuries. CONCLUSION: Hospitals in low-income countries bear a substantial burden of childhood injuries, and systematic surveillance is required to identify the epidemiological distribution of such injuries and understand their risk factors. Methodological standardization for surveillance across countries makes it possible to draw international comparisons and identify common issues. PMID- 19551255 TI - Viet Nam's mandatory motorcycle helmet law and its impact on children. AB - OBJECTIVE: To measure the use of motorcycle helmets in children and to determine the reasons why children wear helmets less often than adults. METHODS: The frequency of helmet wearing among adults and children was ascertained by trained roadside observers, and randomized road user surveys were completed in four major centres in Viet Nam: Hanoi, Ho Chi Minh City, Can Tho and Da Nang. Survey data on key questions were cross tabulated, and chi2 was calculated for significant differences between parents and non-parents (0.05). FINDINGS: The frequency of helmet use in the four study locations ranged from 90-99% among adults, from 15 53% among children 7 but 0.05). Mean HbA1c levels ranged from 8.48 (+/-1.00) to 7.37 (+/-0.99) over time in Group A and from 9.89 (+/-0.86) to 8.34 (+/-1.06) in Group B. The analysis of the HbA1c showed a significant reduction in the first and last 6 months and over the 12 months of the study in two groups (p<0.05). The preprandial scheme demonstrated the largest number and highest percentage of significant drops in HbA1c. CONCLUSIONS: The two monitoring improved the metabolic control and the preprandial scheme was more effective. PMID- 19551273 TI - Factors associated to patients' noncompliance with hypertension treatment. AB - The greatest challenge posed by Systemic Hypertension (SH) is related to patients' compliance with treatment. Thus, this study aimed to determine attendance of these patients to medical appointments and the percentage of adherence to medication and non-medication regimens, and also identify the main reasons hypertensive patients report for non-adherence. This is a descriptive study with 68 hypertensive patients (64.71% women with average age of 63.9 years) at a teaching outpatient clinic. The instruments used for data collection were: multi-professional team care report form, the Morisky-Green test and telephone interview. The results show that 61.76% attended the medical consultations, 86.76% did not comply with the medication regimen and 85.29% did not comply with the non-medication regimen, reporting at least one non-healthy life habit. The emotional factor was the most reported (69.12%) among patients' reasons for non adherence to treatment. The study can support interventions in care delivery to patients with systemic hypertension, with a view to improving their level of adherence and quality of life. PMID- 19551274 TI - Developing a comparative scale of different nociceptive and neuropathic pain through two psychophysical methods. AB - The general aim of this study was to create a comparative scale of different types of pain through different psychophysical methods and different samples. The psychophysical methods used were magnitude estimation and category estimation. The participants were 30 patients from different outpatient clinics, 30 physicians and 30 nurses. The results were: 1) cancer pain, myocardial infarction pain, renal colic, burn-injury pain, and labor pain were considered more intense, regardless of the psychophysical method used or sample studied; 2) The ranking of different pain intensities, comparing the different psychophysical methods used, resulted in significant agreement levels with Kendal values close to 1.00; 3) There were divergences in the perception of the intensities of some types of pain. These divergences were especially observed between professionals and patients. PMID- 19551275 TI - Understanding the dimensions of intensive care: transpersonal caring and complexity theories. AB - This is a descriptive, interpretive and qualitative study carried out at the ICU of a Brazilian teaching hospital. It aimed to understand the dimensions of human caring experienced by health care professionals, clients and their family members at an ICU, based on human caring complexity. The Transpersonal Caring and Complexity theories support theory and data analysis. The following dimensions of care emerged from the themes analyzed according to Ricoeur: self-care, care as an individual value, professional vs. informal care, care as supportive relationship, affective care, humanized care, care as act/attitude, care practice; educative care, dialogical relationship, care coupled to technology, loving care, interactive care, non-care, care ambience, the essence of life and profession, and meaning/purpose of care. We believe in care that encompasses several dimensions presented here, based on the relationship with the other, on the empathetic, sensitive, affectionate, creative, dynamic and understanding being in the totality of the human being. PMID- 19551276 TI - Potential drug interactions in intensive care patients at a teaching hospital. AB - This study assessed potential drugs interactions in intensive care patients at a university hospital in Ceara, northeast Brazil. Of 102 patients studied, 72.5% were exposed to 311 potential drug-drug interactions; 64% of them were females aged 60 years or more and hospital stay was at least 9 days. A statistically significant association was found between number of drugs used and the occurrence of drug interactions. A total of 1,140 drugs were scheduled to be administered concomitantly; of these, 74% had potential for drug interactions. As for the classification of these events, 48.2% had a pharmacokinetic profile; 55.4% were of slow onset; 54.7% had moderate severity; and 60.6% were well-documented in the literature. The most common clinical action taken was 'to monitor signs and symptoms'. Nursing staff can perform 80% of preventive actions to avoid undesirable effects of drug interactions. However, nurses need to have adequate knowledge about drug action mechanisms and triggering factors associated to drug interactions. PMID- 19551277 TI - Factors associated to the development of hypothermia in the intraoperative period. AB - This study aimed to assess factors associated to body temperature changes intraoperatively in patients undergoing elective surgery. A prospective study including 70 patients was carried out in a charity hospital. A data collection instrument was developed and its face and content validity was established. The variables measured were operating room temperature and humidity and patient body temperature at different times. In the multivariate linear regression, the variables type of anesthesia, duration of anesthesia, body mass index, and operating room temperature were directly associated to mean body temperature. Nurses are responsible for planning and implementing effective interventions that can contribute to minimize costs and most importantly reduce hypothermia complications. PMID- 19551278 TI - Nurses' perception of power regarding their clinical role. AB - This article reports on a study on nurses' perception of power regarding their clinical role before and after implementation of a nursing diagnosis classification. Sixty clinical nurses (average age=37.2+/-7.0 years) from a Brazilian teaching hospital answered the Power as Knowing Participation in Change Tool (PKPCT) before and after the implementation of a diagnosis classification. PKPCT has four domains and provides total and partial scores. Reliability coefficients ranged from 0.88 to 0.98. Total scores were not statistically different between assessments (p=0.21), although scores in the 'Involvement in Creating Change' domain were higher in the second assessment (p=0.04). Further studies providing sound evidence regarding the impact of nursing classification systems on nurses' power perception are needed to guide decisions on teaching and clinical practice. PMID- 19551279 TI - Quality management at a hospital's nursing service. AB - The study aimed to know the opinion of nurses in relation to the Quality Management implemented in a hospital service. This is an exploratory and descriptive study carried out with a sample of 17 individuals. The main results, among the 14 principles of quality, reveal that the seventh principle 'adopt and institute leadership' received the highest score. On the other hand, the lowest scored principle was the third: 'cease the dependency of mass inspection'. The obtained results, coupled with theoretical knowledge on the subject and professional experience on the management of nursing services, lead us to the conclusion that nurses consider Total Quality a practical philosophy to be implemented in the services under their responsibility and accept the challenge of overcoming barriers related to tradition, going from discourse to practice. PMID- 19551280 TI - Drug use and risk factors among secondary students. AB - Adolescence is a phase of exposure to several risk behaviors, especially the experimental use of drugs and its associated problems. The study aims to identify risk factors and drug use among secondary students in Comonfort, Guanajuato, Mexico. This is a cross-sectional study, using a version of the Drug User Screening Inventory (DUSI) adapted from Portuguese to Spanish. The sample was composed of 695 (42.9%) students, 52.8% women. Drug use was present in 20.3%, predominantly alcohol and tobacco. Risk factors are related to the male gender, older than 13 years, second and third grades, living with relatives, poor relationships, curiosity, family conflicts, peer pressure and solidarity. CONCLUSION: alcohol and tobacco are the most used drugs and are associated to curiosity and peer pressure. PMID- 19551281 TI - The potential of an instrument to identify social vulnerabilities and health needs: collective health knowledge and practices. AB - OBJECTIVES: To analyze an instrument that aims to understand the health-disease process, its potential to identify social vulnerabilities and health needs to enable the most appropriate interventions in a given territory. PROCEDURES: analysis of the data collection instrument--composed of closed and open questions to identify different profiles of social reproduction (ways of working and living)--used in the Project of Pedagogical Development and activities reported by students at a Collective Health Nursing department. CONCLUSIONS: The instrument enables health care professionals, especially those from Collective Health Nursing, to identify social vulnerabilities and health needs, and guide interventions in different contexts, according to the health promotion perspective. Thus, it reveals potential to transform current health practices, dynamism and constant innovation of these practices in different contexts, according to the 'radicality' that is characteristic of Collective Health. PMID- 19551283 TI - Mothers' health in Brazil and recovering the training of midwives for care in the birth process. AB - The authors briefly analyze the situation of maternal health care in Brazil and, based on their findings, they comment that there have been transformations in the health care model for women and families and propose some measures, including recovery of the work of the obstetric nurse or midwife (obstetriz, in Portuguese). They comment that women care tendencies in the delivery process presuppose incorporation of the paradigm of improving the physiology of valuing women's experience, the approach to the family, health advice that prioritizes prevention, education and relationships, without ignoring safety. They appoint that building this health care model, which includes the work of the midwife, may improve maternal health indicators. PMID- 19551282 TI - Content analysis in studies using the clinical-qualitative method: application and perspectives. AB - Content analysis comprises a set of techniques for organizing communication/information - a procedure used with qualitative data to make themes/topics and concepts/knowledge emerge. Communication content, considering human written or spoken discourse, is complex and presents valuable polysemous characteristic. With the dissemination of the clinical-qualitative method, the use of content analysis, considered an important methodological tool, raises theoretical-practical issues that need to be taken into consideration for its academically precise use. Thus, this article aimed to enumerate specific elements of the content analysis technique and discuss its suitability for the clinical qualitative method that combines generic qualitative methods from Human and Cultural Sciences with the area of Health Sciences. Concepts were selected due to their pertinence, use and eventual adaptation to the method focused on. PMID- 19551284 TI - Blind parents: their experience in care for their children. AB - This study reflects on the difficulties and strategies of blind parents to take care of their children. The situations were related to breastfeeding, bathing, feeding, domestic accidents and administering medication. They use touch, hearing, smelling and the support network, contributing to their autonomy. PMID- 19551285 TI - Animal helminths in human archaeological remains: a review of zoonoses in the past. AB - The authors present a review of records of intestinal parasitic helminths from animals in human archaeological remains, reported since the emergence of paleopathological studies. The objective was to relate paleoparasitological findings to geographic, biotic, and abiotic factors from the environment in which the prehistoric populations lived, and understand some aspects related to the process of human dispersion and biological and cultural evolution. Modification of eating habits and the incorporation of new cultural practices are analyzed from the perspective of zoonoses from prehistory to the present day, especially in Brazilian indigenous populations. Three tables identifying the helminths, their natural hosts, dates, and sites of archaeological findings complete this review. In conclusion, various zoonoses known today have occurred since antiquity, and these data, combined with studies on the emergence and reemergence of diseases, could make possible to compose scenarios for the future. PMID- 19551286 TI - Acellular and "low" pertussis vaccines: adverse events and the role of mutations. AB - OBJECTIVE: to discuss the current PAHO recommendation that does not support the substitution of traditional cellular DTP vaccine by acellular DTP, and the role of mutations, in humans, as the main cause of rare adverse events, such as epileptic-like convulsions, triggered by pertussis vaccine. DATA REVIEW: the main components related to toxic effects of cellular pertussis vaccines are the lipopolysaccharide of bacterial cell wall and pertussis toxin. The removal of part of lipopolysaccharide layer has allowed the creation of a safer cellular pertussis vaccine, with costs comparable to the traditional cellular vaccine, and which may be a substitute for the acellular vaccine. CONCLUSION: The new methodology introduced by Instituto Butantan allows for the development of a new safer pertussis vaccine with low LPS content (Plow), and the use of the lipopolysaccharide obtained in the process in the production of monophosphoryl lipid A. This component has shown potent adjuvant effect when administered together with influenza inactivated vaccine, making possible to reduce the antigen dose, enhancing the production capacity and lowering costs. PMID- 19551287 TI - Risk-factors for non-adherence to antiretroviral therapy. AB - Cross-sectional study analyzed as case-control to identify risk factors for non adherence to antiretroviral therapy. We studied 412 out-clinics HIV infected subjects of three public hospitals of Recife, Pernambuco. The objective was to examine the association between non-adherence to the antiretroviral therapy and biological, social-behavior and demographics and economic factors, factors related to the disease and/or treatment, factors related to life habits and depression symptoms. Variables significantly associated with non-adherence to antiretroviral therapy were: time elapsed since HIV diagnosis (p = 0.002), daily dose (p = 0.046), use of alcohol (p = 0.030) and past drug use (p = 0.048), and borderline p-values were found for educational level (p = 0.093) and family monthly income (p = 0.08). In the multivariable analysis, the factors that remained in the final model were family monthly income, time period with HIV infection and use of alcohol. No association was observed between non-adherence to antiretroviral therapy and gender, age, sexual orientation, marital status, educational level and place of residence. Based on our results and the local situation we suggest: assessment of social needs; training of partners and/or families on supporting adherence, creation of 'adherence groups' to motivate and to reassure patients on the benefits of treatment; counseling and/or psychotherapy for alcohol drinkers. PMID- 19551288 TI - Cytokine and nitric oxide production by mouse macrophages infected with Brazilian flaviviruses. AB - The Flaviviridae family, Flavivirus genus includes viruses that are transmitted to vertebrates by infected mosquitoes or ticks. The genus Flavivirus includes a variety of viruses that cause diseases such as acute febrile illness, encephalitis, and hemorrhagic fever. Flaviviruses primarily infect blood monocytes and tissue macrophages, which have been shown to be permissive, supporting viral replication and serving as virus reservoirs. On the other hand, these cells may have an important antiviral activity related to modulation by cytokine production and by the capacity of these cells to synthesize reactive free radicals such as nitric oxide (NO) which can have a microbicidal effect. The present study was performed in order to determine the production of cytokines interleukin-1beta (IL-1beta), tumor necrosis factor -alpha (TNF-alpha), transforming growth factor- beta (TGF-beta) and interferon -alpha (IFN-alpha) and NO by macrophages infected with one of four Brazilian flaviviruses, Bussuquara virus (BUSV), Yellow Fever virus (YFV), Rocio virus (ROCV) and Encephalitis Saint Louis virus (SLEV), and to verify the possible antiviral effect of NO during macrophage infection with ROCV. Moreover, we asked if the different viruses were able to regulate bacterial lipopolysaccharide (LPS) induced cytokine production. Our results showed that YFV and SLEV reduced the production of IL-1beta and TGF beta by LPS-stimulated macrophages, while ROCV only diminished LPS-stimulated TGF beta synthesis. On the other hand, BUSV more likely favored an enhancement of the LPS-induced production of IL-1beta by macrophages. Additionally, while most of the viruses stimulated the production of IFN-alpha, none of them altered the production of TNF-alpha by murine macrophages. Interestingly, all viruses induced synthesis of NO that was not correlated with antiviral activity for ROCV. PMID- 19551289 TI - Spleen cell proliferation during and after skin myiasis by human bot fly Dermatobia hominis. AB - Spleen cells from mice were examined at 5, 10, 15, 20 and 25 days post-infection (dpi) with Dermatobia hominis larva and at 5, 10, 15, 30 and 60 days post-larval emergence (dple). Cell proliferation in vitro assays were carried out with RPMI 1640 medium and larval secretory product (LSP) of D. hominis at 5, 10, 15, 20 and 25 days. When each group of mice was tested against each medium, significance was only seen for 25 dpi, with increasing order: LSP-10 d, -25 d, -5 d, -20 d, -15 d and RPMI. Significant results were also observed when each medium was tested against mice at each dpi or dple. Each dple group vs. each medium produced significant results only for 10 dple, with increasing order: LSP-5 d, -20 d, -25 d, -10 d, -15 d and RPMI. Comparative tests were also carried out between groups to refine certain observations. The LSPs were also analyzed using SDS-PAGE. The results prove that myiasis caused depletion of spleen cells, particularly under the effect of the LSP-10 and -15, but the cells tended to increase up to 60 dple. This in vitro assay may represent the real systemic immune response in the relationship LSP-D. hominis-host. PMID- 19551290 TI - Giardia intestinalis and nutritional status in children participating in the complementary nutrition program, Antioquia, Colombia, May to October 2006. AB - Giardia intestinalis infection is prevalent throughout the world and widely distributed in developing countries. In general, children display serious consequences to their state of health, including slow height-weight development; therefore, the main aim of this study was to determine the association between Giardia infection and the nutritional status of children who participate in the program of complementary feeding (Mejoramiento Alimentario y Nutricional de Antioquia (MANA) - Instituto Colombiano de Bienestar Familiar (ICBF)). A cross sectional study examining the association of giardiasis with nutritional status was conducted. A total of 2035 children aged eight months to six years-old were studied. Data were collected using structured questionnaires, anthropometric measurements and laboratory analysis of blood and stool samples. Analysis of the results showed that 27.6% of children were infected with G. intestinalis, while 8.1% and 1.9% were mildly and significantly underweight, respectively, and 14.1% presented stunting. Giardiasis was statistically identified as a strong predictor of stunting in this study population. PMID- 19551291 TI - Influence of variables on centrifuge-flotation technique for recovery of Toxocara canis eggs from soil. AB - The purpose of this study was to evaluate the influence of variables in a flotation technique for the recovery of Toxocara canis eggs from soil. The trials were done under standardized conditions on one gram of previously sterilized soil samples contaminated with 200 eggs of T. canis. The following variables were evaluated in serial steps: sieving; type of wash; time of stirring; resuspension of sediment; solution flotation. Centrifuge-flotation in sodium nitrate (d = 1.20 g/cm(3)) was adopted as an initial technique, using Tween 80 (0.2%) and decinormal sodium hydroxide as solutions for washing the samples. Ten tests were done to compare the variables, using counting in triplicate. The sieving of the material reduced significantly the recovery of eggs (p < 0.001) and the number of eggs recovered was higher when the sediment was resuspended (p < 0.05). After standardization, flotation solutions sodium chloride, zinc sulfate, sodium dichromate, magnesium sulfate, and sodium nitrate (d = 1.20g/cm(3)) were compared. The best results were obtained by using zinc sulfate solution. In conclusion, the chances of recovering T. canis eggs from samples using flotation solutions can be increased by washing of soil twice using distilled water, and resuspension of sediment. On the other hand, the sieving procedure can drastically reduce the number of eggs. PMID- 19551292 TI - Seabather's eruption: a clinical and epidemiological study of 38 cases in Santa Catarina State, Brazil. AB - Seabather's eruption (SBE) is an intensely itchy, papule-erythematous dermatitis that occurs predominantly in regions of the body covered by bathing costumes, after exposure to marine water. The planulae larvae of Linuche unguiculata scyphomedusae (thimble jellyfish) are the etiologic agent of the dermatitis, which is frequent in waters of Caribbean, Gulf of Mexico and Florida. The authors report 38 cases of SBE in the State of Santa Catarina (Southern region of Brazil), with emphasis on their clinical and epidemiological aspects, such as profile of victim, topography of the papules and conditions predisposing to the accident. PMID- 19551293 TI - Clinical and therapeutic applications of Nigella in the management of tropical infectious disorders besides Schistosomiasis. PMID- 19551294 TI - Molecular characterization of toxins in Staphylococcus aureus isolated from milk and 'coalho' cheese in cities from the Agreste region of Pernambuco. PMID- 19551295 TI - Resection of a giant malignant mediastinal peripheral nerve sheath tumour under cardiopulmonary bypass. AB - Malignant peripheral nerve sheath tumour (MPNST) is a rare occurrence in the mediastinum. It is biologically aggressive and is generally resistant to chemoradiation therapy. The mainstay of treatment is complete surgical resection. We report a large MPNST which invaded into the adjacent aortic wall in a 50-year old man. Extensive resection, which included aortic reconstruction under cardiopulmonary bypass and deep hypothermic arrest, was necessary for a good long term outcome. PMID- 19551296 TI - Gastric adenocarcinoma occurring in a young patient with common variable immunodeficiency syndrome. AB - Patients with common variable immunodeficiency syndrome (CVID) have an increased risk of gastric adenocarcinoma. We describe a case of gastric adenocarcinoma in a 29-year-old man with CVID. He complained of dyspepsia and weight loss. Endoscopy showed an antral lesion. He underwent subtotal gastrectomy with postoperative adjuvant chemoradiation, and remained disease-free for three years. CVID is a predisposing factor for gastric adenocarcinoma. Gastric complaints are common among these patients and should be viewed seriously. Endoscopy is performed to detect any pathology. Premalignant conditions like chronic atrophic gastritis, intestinal metaplasia and dysplasia require regular endoscopic surveillance in these high-risk patients. PMID- 19551297 TI - Tenosynovial giant cell tumour of the posterior cruciate ligament and its arthroscopic treatment. AB - Tenosynovial giant cell tumours originate from synovial tissues of the joints, tendon sheaths, mucosal bursas or fibrous tissues adjacent to tendons. Tenosynovial giant cell tumours are rarely intra-articular. We report a giant cell tumour of the tendon sheath arising from the posterior cruciate ligament diagnosed by magnetic resonance imaging and resected arthroscopically in a 54 year-old woman. PMID- 19551298 TI - Open simultaneous double dislocation of the metacarpophalangeal and interphalangeal joints of the thumb with ulnar collateral ligament avulsion. AB - Double dislocations of the thumb at the metacarpophalangeal joint (MCPJ) and interphalangeal joint (IPJ) occurring at the same time are seldom seen in medical practice. The sporadic occurrence of this interesting injury makes it difficult to formulate a clear management protocol. We report an unusual combination of injury involving the right thumb of a 60-year-old man, where the IPJ was dislocated dorsally with a volar dislocation of the MCPJ. It was an open injury with avulsion of the ulnar collateral ligament of the MCPJ. The dislocation was treated successfully by closed reduction in the casualty department, under a ring block. The ulnar collateral ligament was repaired later in the operating theatre. At six months' follow-up, the patient had a full painless range of movement in his thumb with no instability. To our knowledge, this is the first reported case of a thumb double dislocation where the injury was open at both joints. This is also the first case where a volar dislocation of the MCPJ was treated successfully by closed reduction, with an excellent functional outcome. PMID- 19551299 TI - Double inferior vena cava: a report of three cases. AB - Double inferior vena cava (IVC) is a congenital anomaly resulting from the persistence of the embryonic venous system. The majority of cases are clinically silent and diagnosed incidentally on imaging for other reasons. However, these venous anomalies may have significant clinical implications, especially during retroperitoneal surgery and in the treatment of thromboembolic diseases. We report three cases of double IVC and review the relevant literature. The clinical importance of recognising double IVC is discussed. PMID- 19551300 TI - Bilateral coracoclavicular joints as a rare cause of bilateral thoracic outlet syndrome and shoulder pain treated successfully by conservative means. AB - The coracoclavicular joint (CCJ) is a rare anomalous joint. Symptomatic CCJ, being an exceptional rarity, makes it difficult to formulate a standard set of practice or guidelines. We report a 50-year-old Indian man, a machine operator by profession, who experienced bilateral shoulder pain and arm paraesthesia for two years, and was diagnosed with bilateral CCJ. The symptoms gradually increased, affecting his daily activities. Dynamic magnetic resonance imaging revealed the compression of the brachial plexus in extreme shoulder abduction. After a thorough search of the literature, we retrieved four similar cases, all of them treated with individualised protocols. None of the cases was bilateral. The lack of clear evidence in any particular direction and the patient's medical condition prompted us to give a conservative trial, before embarking on more invasive methods. He showed rapid response to the conservative treatment with remission of all symptoms. To the best of our knowledge, this is the first reported case of bilateral symptomatic CCJ with bilateral thoracic outlet syndrome, that was managed conservatively. PMID- 19551301 TI - Recurrent pancreatitis secondary to pancreatic ascariasis. AB - Ascaris lumbricoides infestations are endemic in tropical countries. Ascaris lumbricoides can occasionally cause biliary obstruction and result in obstructive jaundice or pancreatitis. We present a 34-year-old Bangladeshi woman with biliary ascariasis, resulting in recurrent pancreatitis. Her diagnosis was made with endoscopic retrograde cholangiopancreatography performed during an acute attack of pain. PMID- 19551302 TI - Coloduodenal fistula: a rare complication of right-sided diverticulitis. AB - Coloduodenal fistula is an uncommon disease entity. It can be caused by either a malignant or benign disease. We report the fifth case of coloduodenal fistula secondary to colonic diverticulosis. Our patient, a 60-year-old man, presented with severe diarrhoea and recurrent severe hypokalaemia. A partial colectomy with en bloc excision of the fistula was performed, and the duodenal defect was closed primarily. A literature review was carried out on the aetiology, presentation, diagnosis and management of coloduodenal fistula. PMID- 19551303 TI - Gigantism of the lower limb in Klippel-Trenaunay syndrome: anatomy of the lateral marginal vein. AB - The Klippel-Trenaunay syndrome is a combination of venous and capillary malformations associated with soft tissue and/or bony limb hypertrophy, with or without lymphatic malformations. Although persistent foetal veins are rare, the persistence of the lateral marginal vein is a common association in this syndrome. It results in venous hypertension, which gives rise to venous varicosities, which are commonly seen in this syndrome. This is a case report of a 28-year-old man with Klippel-Trenaunay syndrome, with persistence of the lateral marginal vein, affecting his right lower limb. He was treated with an above-knee amputation. The amputated limb was dissected to demonstrate the anatomy of the lateral marginal vein. To the best of the authors' knowledge, the gross anatomy of the lateral marginal vein has not been previously reported. PMID- 19551304 TI - Primary diaphyseal tuberculosis of the tibia. AB - Tuberculous osteomyelitis involving primarily the diaphysis without articular involvement is very rare. Pain and swelling are the common presenting symptoms. The nonspecific nature of the symptoms leads to a delay in the diagnosis. Radiographs may mimic pyogenic osteomyelitis, Brodie's abscess, tumours or granulomatous lesions. Curettage of the lesion and the histopathological examination of the material obtained are necessary for confirmation of the diagnosis and offer a chance for early healing. We report the successful diagnosis and management of a rare case in a 28-year-old man of Indian origin afflicted with primary diaphyseal tuberculosis of the left tibia. PMID- 19551305 TI - Responding to the new influenza A (H1N1) 2009 pandemic: moving forward together. PMID- 19551306 TI - The evolution of the anatomically modern or advanced Homo sapiens: time, place, process, affinities and variations. AB - This paper surveys the opinions expressed in the recent literature on the origins of the anatomically- modern Homo sapiens, and reviews the evidence from cranial and dental morphology argued by proponents of opposing views to support their case. It also critically analyses problems facing the interpretation of the evidence in arriving at a definitive conclusion to the debate. PMID- 19551307 TI - Authorship and acknowledgements. AB - All persons designated as authors of a scientific manuscript should qualify for authorship, and all those who qualify should be listed. Each author should have participated sufficiently in the work to take public responsibility for appropriate portions of the content, and have made substantial intellectual contribution to the submitted manuscript. All contributors who do not meet the criteria for authorship should be listed in the acknowledgements section. Ghost authors are undesirable while professional medical writers are helpful to authors who are not native English speakers, have language difficulties, or lack training or expertise in medical writing. PMID- 19551309 TI - New influenza A (H1N1) 2009 in Singapore: the first ten adult imported cases. AB - INTRODUCTION: Since late March 2009, a novel influenza H1N1 strain emerged in humans in Mexico and the United States. It has rapidly spread to many countries on different continents, prompting unprecedented activation of pandemic preparedness plans. Singapore has adopted a containment strategy with active screening of febrile travellers with respiratory symptoms from affected countries since April 27, 2009. METHODS: All cases with new influenza A (H1N1) confirmed on polymerase chain reaction assay on combined nasal and throat swabs and who were admitted to the Communicable Disease Centre, were included in a prospective evaluation of clinical characteristics of new influenza A (H1N1). RESULTS: From May 26 to June 3, 2009, there were ten patients with a mean age of 27.6 years, seven of whom were female. All but one travelled from the United States, six of whom travelled from New York; the last one travelled from the Philippines. Clinical illness developed within a mean of 1.4 days after arrival in Singapore, and presentation to the emergency department at a mean of 2.7 days from illness onset. Fever occurred in 90 percent, cough 70 percent, coryza 40 percent, sore throat and myalgia/arthralgia 30 percent; none had diarrhoea. The fever lasted a mean of 2.1 days. All were treated with oseltamivir. The clinical course was uncomplicated in all cases. CONCLUSION: Clinical features of new influenza A (H1N1) appeared mild, and ran an uncomplicated course in immunocompetent patients. PMID- 19551308 TI - Management of novel influenza epidemics in Singapore: consensus recommendations from the Hospital Influenza Workgroup (Singapore). AB - The recent emergence and global spread of a novel strain of influenza A (H1N1) virus has resulted in the first influenza pandemic of the 21st century. With its rapid spread to more than 70 countries within three months, governments are faced with the challenge of either containing or mitigating this influenza pandemic. The aim of this paper is to provide evidence-based consensus recommendations in the areas of infection control, antiviral treatment, chemoprophylaxis, antibiotic stockpiling and vaccination to guide decision-making for clinicians and administrators within the Singapore context. As the transmissibility and virulence of this new influenza A (H1N1) virus may evolve over time, we have tailored our recommendations according to several potential scenarios of viral virulence and transmissibility. PMID- 19551310 TI - Foetal peak systolic velocity in the middle cerebral artery: an Asian reference range. AB - INTRODUCTION: The aim of this study was to establish reference values of peak systolic blood flow velocity measurement in the foetal middle cerebral artery (MCA-PSV) in the local Asian obstetric population and to compare our reference ranges with those of previously-published studies. METHODS: 329 normal pregnant women attending the outpatient antenatal clinics of the Department of Obstetrics and Gynaecology in the Singapore General Hospital underwent Doppler ultrasonography at least once between 16 and 40 weeks' gestation. The blood flow velocity recordings from the foetal middle cerebral artery were obtained. New reference ranges were constructed by regressing each parameter on gestational age. RESULTS: New reference ranges for foetal middle cerebral artery with gestation were constructed for an Asian population. Our reference curves were compared with that of a previously-constructed one. CONCLUSION: MCA-PSV increases with advancing gestational age. There appear to be differences between Asian and non-Asian reference ranges for MCA-PSV. PMID- 19551311 TI - Pattern of Turner syndrome in Singapore (1999-2004). AB - INTRODUCTION: Turner syndrome is the most common sex chromosomal abnormality in female foetuses, and is associated with a high proportion of cardiac anomalies. The aim of this study was to look at the incidence, demographical data and epidemiological pattern of Turner syndrome in Singapore from 1999 to 2004 and to examine the birth defects associated with this condition, specifically with reference to cardiac defects. METHODS: Data on Turner syndrome cases born in 1999 2004 were retrieved from the National Birth Defects Registry (NBDR) and analysed. Data on congenital cardiac defect cases notified to the NBDR in the same time period were also retrieved and compared with the Turner syndrome cases. RESULTS: There were a total of 101 cases of Turner syndrome in the six-year period from 1999-2004, yielding an overall incidence of 0.85 per 1,000 female live births, or one in 1,180 female live births. The incidence was lowest among Indians (0.38 per 1,000) compared to Malays (0.72 per 1,000) and Chinese (0.90 per 1,000). 75 cases (74.3 percent) had the 45,X karyotype, while the other 26 cases (25.7 percent) were mosaics. The mean maternal age for 45,X was lower (32.2 years, range 22-42) compared to mosaics (34.5 years, range 27-40). 19.8 percent (20/101) were live births, 38.6 percent (39/101) were terminated pregnancies and 41.6 percent (42/101) were spontaneous miscarriages. 13.9 percent of Turner syndrome babies had cardiac defects compared to 1.2 percent in the general population (p-value is less than 0.0001). Major cardiac defects found among Turner syndrome babies compared to the general population included the coarctation of the aorta (5.9 percent compared to 0.03 percent, p-value is less than 0.0001), atrial septal defects (3.0 percent compared to 0.6 percent, p-value is 0.006), a hypoplastic left heart (2.0 percent compared to 0.05 percent, p-value is less than 0.0001), aortic hypoplasia (3.0 percent compared to 0.01 percent, p-value is less than 0.0001) and dextrocardia (1.0 percent compared to 0.02 percent, p-value is 0.0002). CONCLUSION: Cardiac defects, particularly left-sided ones, are significantly more common among Turner syndrome foetuses. The true incidence of this syndrome is likely to be higher than that quoted in this study, and can only be solved when a complete screening of an entire population has been performed. PMID- 19551312 TI - Endovenous laser treatment for varicose veins in Singapore: a single centre experience of 169 patients over two years. AB - INTRODUCTION: Endovenous laser therapy (EVLT) is one of the many minimally invasive procedures that have been developed in recent years for the treatment of varicose veins. We present our single centre experience of 169 patients who underwent EVLT. METHODS: All patients who underwent EVLT since its introduction in our institution were included in our series. RESULTS: A total of 270 incompetent long saphenous veins in 169 patients were ablated by EVLT from February 2006 to January 2008. Bilateral EVLT was performed in 101 (59.8 percent) patients, with the remaining 68 (40.2 percent) undergoing unilateral EVLT. The mean age of the patients was 54 (range 19-78) years and there were 112 (66.3 percent) women. The majority of our patients (63.3 percent) had symptoms for more than five years. The symptoms included lower limb cramps and aches (47.9 percent) as well as lower limb swelling (16.6 percent). The median follow-up was six months. Complications from our series included numbness over the affected lower limbs (10.7 percent) and skin pigmentation (4.1 percent). Only 2.4 percent of patients had recurrence after one year. CONCLUSION: Early results with EVLT have been impressive, and this study has reaffirmed the safety and effectiveness of EVLT in the treatment of varicose veins. PMID- 19551313 TI - Is noninvasive pressure support ventilation as effective and safe as continuous positive airway pressure in cardiogenic pulmonary oedema? AB - INTRODUCTION: Noninvasive ventilation (NIV) with continuous positive airway pressure (CPAP) has been shown to decrease endotracheal intubation and mortality in patients with acute cardiogenic pulmonary oedema (ACPE). The Three Interventions in Cardiogenic Pulmonary Oedema showed no advantage of NIV over standard medical therapy. This meta-analysis is an update on the efficacy and safety of two different forms of NIV (noninvasive pressure support ventilation [NIPSV] vs. CPAP) in patients with ACPE. METHODS: We searched the MEDLINE and EMBASE databases for randomised clinical trials published from 1980 to 2008 that have compared NIPSV and CPAP in patients with ACPE. We calculated the odds ratio (OR) with 95 percent confidence intervals (CI) and pooled the results using three different statistical models (fixed effects, random effects and exact method). RESULTS: Ten studies (577 and 576 in the CPAP and NIPSV groups, respectively) met our inclusion criteria. NIPSV performed similar to CPAP in decreasing the intubation rates (OR 0.8; 95 percent CI 0.43-1.49), hospital mortality (OR 1.08; 95 percent CI 0.76-1.54) and the occurrence of myocardial infarction (OR 0.8; 95 percent CI 0.36-1.76). The results were similar when pooling the data with any of the three statistical methods and stratifying for the type of pressure therapy (fixed vs. variable) except for myocardial infarction, which was more frequent in the fixed pressure NIPSV arm (OR 5.06; 95 percent CI 1.66-15.44). CONCLUSION: NIPSV appears to be as safe and efficacious as CPAP, if titrated rather than fixed pressures are employed. PMID- 19551314 TI - Role of fibrinolytic markers in acute stroke. AB - INTRODUCTION: The fibrinolytic system plays an important role in normal haemostasis and endothelial function. This study was conducted to compare three fibrinolytic markers, i.e. plasminogen, tissue-plasminogen activator (t-PA) and plasminogen activator inhibitor type-1 (PAI-1) between acute stroke and stable non-stroke patients and to investigate the clinical significance of these markers. METHODS: A prospective study was done for a one-year period upon obtaining ethical approval from the local institution. 106 non-stroke individuals from general outpatient clinics (control group) and 51 acute stroke patients were selected. All subjects were tested for t-PA and PAI-1 levels using the enzyme immunoassay technique (Biopool TintElize) and for plasminogen level by colorimetric assay (HemosIL). They were followed up over a period of three months for survival and neurological recovery. RESULTS: Only the mean t-PA level was significantly higher in acute stroke patients compared to the control group, including after adjusting for confounders (using ANCOVA). There was no statistical association between the three fibrinolytic markers and the age, gender, stroke subtypes, number of risk factors, functional impairment, survival and neurological recovery. We observed that all the eight patients who died within one month of stroke onset had high levels of t-PA. An association between high t-PA antigen and acute stroke was found during a cerebrovascular event with a 4.6-fold odds ratio compared to non-stroke controls. CONCLUSION: High t-PA antigen in acute stroke patients probably indicates a poor prognosis. Its value as a marker for monitoring and prognostication needs to be evaluated as a routine clinical practice. PMID- 19551315 TI - The association between serum apolipoprotein A-I and apolipoprotein B and the severity of angiographical coronary artery disease. AB - INTRODUCTION: The aim of this study was to investigate the relationship between serum apolipoprotein A-I (apoA-I) and apolipoprotein B (apoB) and the severity of coronary artery stenosis. METHODS: This case-control study was carried out on 106 patients who underwent angiography and 100 healthy controls. ApoA-I and apoB as well as the serum total cholesterol, high-density lipoprotein cholesterol (HDL C), triglyceride and low-density lipoprotein cholesterol (LDL-C) levels were measured. Very low-density lipoprotein cholesterol levels and the LDL-C/HDL-C ratio were calculated. RESULTS: In an Iranian population with coronary artery disease (79 men and 27 women, aged 53 +/- 8.5 years), the increased levels of apoA-I and apoB were correlated with the number of involved vessels and the severity of coronary lesions. However, no significant correlation was found between the serum values of lipids as well as other lipoproteins and the number of vessels involved and the severity of coronary lesions. CONCLUSION: ApoA-I and apoB are indicated as risk factors for cardiovascular and, possibly, cerebrovascular diseases. From this study, it may be concluded that apoA-I and apoB serum concentration levels are independent risk factors for coronary atherosclerosis in the Iranian population. It also demonstrates a direct relationship between the severity of coronary atherosclerosis and the number of lesions in the involved vessels. It can be regarded as an index for the relationship of apoA-I and apoB to the early, still clinically asymptomatic, steps of the pathogenesis of coronary disease. PMID- 19551316 TI - Quantitative assessment of posture stability using computerised dynamic posturography in type 2 diabetic patients with neuropathy and its relation to glycaemic control. AB - INTRODUCTION: Patients with diabetic neuropathy have an imbalance, which comes with a higher risk of falls. The aim of this study was to assess posture stability using computerised dynamic posturography in type 2 diabetics mellitus patients with neuropathy as well as its relation to glycaemic control. METHODS: 54 type 2 diabetics mellitus patients with peripheral neuropathy were recruited, together with 18 type 2 diabetics mellitus patients without peripheral neuropathy acting as the control group. The first group was divided into two subgroups according to glycaemic control assessed by HbA1c (A1c), the first subgroup comprising 24 patients had good glycaemic control with A1c less than or equal to seven percent and the second subgroup with 30 patients had poor glycaemic control with A1c more than 7 percent. The postural stability was evaluated using dynamic posturography. RESULTS: The composite equilibrium score, sensory organisation test 1, 2 and 3 conditions were significantly lower in the neuropathic group as compared to the non-neuropathic group (p-value is less than 0.001). A1c was significantly correlated with the composite equilibrium score in the neuropathic group with poor glycaemic control (r-value equal to -0.395) but not correlated in the neuropathic group with good glycaemic control (r-value equal to 0.151). CONCLUSION: Posture instability in type 2 diabetic patients with peripheral neuropathy reflects an impairment of the somatosensory system; also, poor glycaemic control resulted in more posture instability. The early detection of imbalance using dynamic posturography and achieving good glycaemic control may be of great help in the prevention of falls in such patients. PMID- 19551317 TI - Impact of Ramadan on upper gastrointestinal endoscopy referrals in Brunei Darussalam. AB - INTRODUCTION: Fasting during the month of Ramadan is one of the five holy pillars in the Islamic faith and is an obligation for all its followers. Prolonged fasting may precipitate or exacerbate gastrointestinal (GI) complaints. This study assessed the impact of Ramadan on referrals for upper GI endoscopy in a tertiary referral centre. METHODS: 1,661 patients referred to the centre a month before, during and a month after the month of Ramadan over a four-year period (2004-07) were retrospectively studied. Significant endoscopic findings were taken as any bleeding lesions, severe or complicated oesophagitis, peptic ulcer disease, portal hypertension related pathologies and malignancies. RESULTS: Overall, there was significantly less workload generated during the fasting month (397 patients, 5.7 +/- 2.7 cases per list) compared to before (603 patients, 6.9 +/- 3.0 cases per list, p-value is equal to 0.036) and after (661 patients, 7.8 +/- 3.4 cases per list, p-value is less than 0.001) the fasting month. There was no significant difference between the period before and after the fasting month (p-value equal to 0.124). There were no significant differences in the mean age and gender of the patients. During the fasting month, there were differences in the ethnicity (fewer Malays and more Chinese, p-value is equal to 0.002) and referral sources (more wards and fewer clinics, p-value is less than 0.001). There were no differences in the referral indications, oesophageal and gastric findings, but there was a significant difference in the duodenal findings (p value is equal to 0.001), especially ulcer disease, during the fasting month. CONCLUSION: This study showed that significantly less workload was generated during the fasting month of Ramadan compared to the non-fasting months. There were also some differences in the referral sources, ethnicity and the endoscopic findings. PMID- 19551318 TI - Antioxidant status and smoking habits: relationship with diet. AB - INTRODUCTION: The present study was conducted to assess the association between smoking, dietary intake of antioxidants and plasma indices of oxidative stress and antioxidant defences in male smokers (cigarette and bidi smokers). METHODS: The study sample consisted of 100 healthy men, including 50 non-smokers and 50 smokers, who were subclassified into 25 cigarette smokers and 25 bidi smokers, aged 18-55 years. Erythrocyte superoxide dismutase and plasma ascorbic acid were measured as antioxidants and erythrocyte malondialdehyde as an oxidative stress index, by colorimetric methods. RESULTS: Smokers ate less fruits and vegetables than non-smokers, leading to them having a lower antioxidant level. Erythrocyte superoxide dismutase was significantly lower in cigarette smokers (0.193 U/mgP, p value is less than 0.05) and bidi smokers (0.169 U/mgP, p-value is less than 0.001) as compared to non-smokers (0.231 U/mgP). Plasma ascorbic acid was also significantly lower in cigarette smokers (1.45 mg/100 ml, p-value is less than 0.05) as well as in bidi smokers (1.38 mg/100 ml, p-value is less than 0.001) as compared to non-smokers (1.73 mg/100 ml). There was a significant increase in erythrocyte malondialdehyde concentration levels in cigarette smokers (171.47 micromol/gHb, p-value is less than 0.05) as well as in bidi smokers (231.04 micromol/gHb, p-value is less than 0.001) as compared to non-smokers (127.30 micromol/gHb). CONCLUSION: These results provide enough evidence of increased oxidative stress and a compromised antioxidant defence system in smokers, and they are more profound in bidi smokers than in those smoking cigarettes. This study also revealed that the diet and nutrient intake of smokers are different from that of non-smokers. PMID- 19551319 TI - Ertapenem susceptibility of extended spectrum beta-lactamase-producing Enterobacteriaceae at a tertiary care centre in India. AB - INTRODUCTION: Infections caused by multidrug-resistant organisms such as extended spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have come to assume widespread proportions. Carbapenems (imipenem and meropenem) are the drugs of choice for the treatment of infections caused by ESBL-producing organisms. There is limited clinical data regarding the efficacy of the latest carbapenem, called ertapenem, against these organisms in the Indian subcontinent. In this study, ertapenem susceptibility in ESBL-producing clinical isolates was evaluated. The in vitro activities of the three carbapenems were compared in ertapenem-resistant isolates. METHODS: A total of 205 ESBL-producing Enterobacteriaceae collected from inpatients and outpatients at the All India Institute of Medical Sciences, were identified and tested for antimicrobial susceptibility by the disc diffusion method and Vitek 2 advanced expert system. Ertapenem susceptibility was performed by disc diffusion and Vitek 2 in all the isolates and by E-test in 100 isolates. RESULTS: 191 (93 percent) of the ESBL-producing isolates tested were susceptible to ertapenem. All ertapenem-susceptible isolates were also susceptible to imipenem and meropenem. Isolates with low-level ertapenem resistance retained their susceptibility to imipenem and meropenem, whereas those with high-level ertapenem resistance were resistant to both imipenem and meropenem. CONCLUSION: Our results suggest that ertapenem may be a viable alternative to other carbapenems for the treatment of infections caused by ESBL-producing clinical isolates. Clinical outcome studies are required to determine if ertapenem is effective for the treatment of infections caused by these organisms. PMID- 19551320 TI - Gastrointestinal tuberculosis. AB - Tuberculosis (TB) infection is still common and remains an important cause of morbidity and mortality, particularly in underdeveloped and developing nations. The gastrointestinal (GI) tract is the sixth commonest extrapulmonary site to be affected after lymphatic, genitourinary, bones and joints, miliary and meningeal involvement. Manifestations can be non-specific and mimic many conditions, including malignancies. Findings from endoscopy and radiological imaging are myriad, and depend on the stage of the disease and the time at which investigations are carried out. Hence, diagnosis can be difficult. Endoscopy is now the investigation of choice as it allows for visualisation and the sampling of tissue for histology and culture. This is complemented by radiological imaging. This pictorial essay reviews some of the endoscopic and radiological findings of non-human immunodeficiency virus associated proven GI TB infections that we have encountered in a tertiary referral centre. PMID- 19551321 TI - CT findings of tuberculous peritonitis. PMID- 19551322 TI - "Fin de siecle"--how we got into this financial and economic crisis and what implications it has for the practice of medicine in Singapore. PMID- 19551325 TI - Oocytes and early embryos selectively express the survival factor BCL2L10. AB - Apoptosis has been reported in oocytes and human preimplantation embryos both in vitro and in vivo. BCL-2 family proteins are likely to play a pivotal role in controlling oocyte and early embryo degeneration. However, no BCL-2-related survival factors have been identified that would specifically function during oocyte maturation, after fertilization and during early embryogenesis. Here, we performed a comprehensive tissue expression pattern analysis of the BCL-2 family at the mRNA level. While expression of various members was detected in human oocytes and during early primate embryogenesis, our data indicate that BCL2L10 is the predominant maternally loaded Bcl-2 family transcript, revealing an evolutionary conserved expression profile at the egg-to-zygote transition. We provide evidence that BCL2L10 is associated with the microtubule binding protein translationally controlled tumor protein and mitochondria, with a stage-specific redistribution along the pericortical regulatory ooplasm. In dying oocytes, BCL2L10 colocalized with proapoptotic BAX and neutralization of BCL2L10 accelerated oocyte death. We propose BCL2L10 as a novel and prime candidate related to oocyte maturation, fertility, and embryo developmental competence. PMID- 19551326 TI - The prevalence of autism spectrum disorders: impact of diagnostic instrument and non-response bias. AB - BACKGROUND: A large part of the variability in rates of autism spectrum disorders (ASD) across studies is non-aetiologic, and can be explained by differences in diagnostic criteria, case-finding method, and other issues of study design. AIM: To investigate the effects on ASD prevalence of two methodological issues; non response bias and case ascertainment. We compared the findings of using a semi structured parent interview versus in-depth clinical assessment, including an ASD specific interview. We further explored whether including information on non responders affected the ASD prevalence estimate. METHOD: A total population of 7- to 9-year olds (N = 9,430) was screened for ASD with the autism spectrum screening questionnaire (ASSQ) in the Bergen Child Study (BCS). Children scoring above the 98th percentile on parent and/or teacher ASSQ were invited to participate in the second and subsequently in the third phase of the BCS where they were assessed for ASD using the Development and Well-Being Assessment (DAWBA), and the Diagnostic Interview for Social and Communication disorders (DISCO), respectively. RESULTS: Clinical assessment using DISCO confirmed all DAWBA ASD cases, but also diagnosed additional cases. DISCO-generated minimum prevalence for ASD was 0.21%, whereas estimated prevalence was 0.72%, increasing to 0.87% when adjusting for non-responders. The DAWBA estimate for the same population was 0.44%. CONCLUSION: Large variances in prevalence rates across studies can be explained by methodological differences. Both information about assessment method and non-response are crucial when interpreting prevalence rates of ASD. PMID- 19551327 TI - Polygamy and mental health of adolescents. AB - The objective is to study the influence of polygamous versus monogamous marriage on the mental health of adolescents in an Israeli Bedouin population. Pupils aged 11-18 years attending schools in Bedouin Arab communities in southern Israel were asked to complete a demographic questionnaire and a panel of psychological instruments measuring competence and behavioral problems, anxiety and depression. Findings were compared between pupils of families with one wife and pupils of families with more than one wife. The population comprised 406 pupils of mean age 14.5 years; 56% were female. Fifty-three percent were from polygamous marriages and 47% from monogamous marriages. After allowing for the influence of socioeconomic factors, there were no differences between offspring of polygamous marriages and those of monogamous marriages for any of the psychological scales. When polygamy is the accepted practice in a particular social milieu, it does not have a deleterious psychological effect on adolescents. PMID- 19551328 TI - Breast milk excretion Kinetic of b-HCH, pp'DDE and pp'DDT. AB - Breast milk is considered the most important route in the elimination of deposited organochlorine pesticides in a mother's body. The equilibrium of organochlorine pesticides in the human body considers the elements of internal transport processes, the equilibrium pattern between pesticides and tissue fat contents, and the mobilization of lipids and lipoproteins among body parts. The aim of this study was to determine organochlorine pesticide levels in breast milk samples from the 4th to the 30th day of lactation and the trend in their concentration time so as to forecast the time tendency of residue levels and the pesticide excretion pattern. Milk samples were taken from forty participants and analyzed by GLC-ECD. The organochlorine pesticide residues determined in the breast milk samples during lactation decreased: beta-HCH from 0.095 to 0.066 mg/kg, pp'DDE from 1.807 to 1.423 mg/kg and pp'DDT from 0.528 to 0.405 mg/kg, at the characteristic rate for each compound. The obtained results compared with the calculated fits of forecasts were parallel and did not exhibit significant differences. The newborn baby exposed during lactation had organochlorine pesticide residues whose levels decreased permanently. The levels depended not only on the breast milk nutrition, but also on the total environmental exposures which included air pollution as a significant contamination source. PMID- 19551329 TI - Interference of shot noise of open-channel current with analysis of fast gating: patchers do not (Yet) have to care. AB - Microsecond gating of ion channels can be evaluated by fitting beta distributions to amplitude histograms of measured time series. The shape of these histograms is determined not only by the rate constants of the gating process (in relation to the filter frequency) but also by baseline noise and shot noise, resulting from the stochastic nature of ion flow. Under normal temporal resolution, the small shot noise can be ignored. This simplification may no longer be legitimate when rate constants reach the range above 1 mus(-1). Here, the influence of shot noise is studied by means of simulated time series for several values of single-channel current of the fully open state and baseline noise. Under realistic optimal conditions (16 pA current, 1 pA noise, 50 kHz bandwidth), ignoring the shot noise leads to an underestimation of the rate constants above 1 mus(-1) by a factor of about 2.5. However, in that range, the scatter of the evaluated rate constants is at least of the same magnitude, obscuring the systematic error. The incorporation of shot noise into the analysis will become more important when amplifiers with significantly reduced noise become available. PMID- 19551331 TI - Endoscopic papillary large balloon dilation for large common bile duct stones. AB - Endoscopic papillary large balloon dilation (EPLBD) involves endoscopic biliary sphincterotomy (EBS) followed by balloon dilation using a 12-20-mm balloon to remove large or difficult stones from the common bile duct. The complications and limitations of endoscopic biliary sphincterotomy (EBS) are well known. Endoscopic papillary balloon dilation (EPBD) with a smaller diameter balloon but without sphincterotomy is widely used in a number of regions of the world for removal of routine bile duct stones and has been investigated as an alternative to EBS. EPBD, however, appears to be associated with an increased risk of pancreatitis. EPLBD differs from EPBD as it involves EBS followed by large balloon dilation. EPLBD would theoretically combine advantages of sphincterotomy and balloon dilation by increasing efficacy at stone extraction while minimizing complications of both EBS and EBD. A review of the available literature for EPBLD shows that it is relatively safe and effective. A high success rate (up to 95%) has been described for stone removal using EPLBD, with a low complication rate. Unlike EPBD, EBLBD does not appear to be associated with a higher risk of post ERCP pancreatitis, probably because of separation of the biliary and pancreatic sphincters after EBS. EPLBD appears to be a reasonable option for removal of large or difficult common bile duct stones. This technique may be especially helpful in patients with difficult papillary anatomy, such as those with small papillae, intra- or peri-diverticular papilla. Its role in patients with coagulopathy or other risks for bleeding remains to be investigated. PMID- 19551332 TI - Covered metallic stents in the management of malignant and benign pancreatobiliary strictures. AB - In the endoscopic management of unresectable malignant biliary obstructions by placement of a metallic stent (MS), longer patency and a lower incidence of stent occlusion are desirable goals. With its mesh structure, the uncovered MS (UMS) is occluded mainly by tumor or tissue ingrowth, making it impossible to remove. The covered MS (CMS) was developed to overcome these disadvantages, and was shown to maintain patency longer than the UMS in our randomized study. The most important characteristic of the CMS is that it is removable, allowing it to be used in patients with resectable malignancies and benign strictures. In addition, the drug-eluting CMS provides an additional approach to the treatment of biliary malignancies. The CMS may also change the treatment paradigm for biliary strictures and strictures due to chronic pancreatitis. The CMS is analogous to a large-bore, expandable plastic stent and is effective both as an endoprosthesis and a dilating or anti-cancer device. However, to better understand the utility of these devices, we need to first consider mechanical properties such as radial force (RF, expansion force) and axial force (AF, straightening force). AF is particularly important when developing CMSs because of related complications. PMID- 19551333 TI - Endotherapy for pancreatic necrosis and abscess: endoscopic drainage and necrosectomy. AB - Pancreatic necrosis and abscess are among the most severe complications of acute pancreatitis. Endoscopic drainage of pancreatic fluid collections has been increasingly performed in many tertiary care centers. The type of fluid collection that is being intervened upon determines the outcome. The development of endoscopic ultrasonography (EUS) has expanded the safety and efficacy of this modality by allowing one to access and drain more challenging fluid collections. The technique and review of current literature regarding endoscopic therapy of pancreatic necrosis and abscess will be discussed. PMID- 19551334 TI - Multiple lytic lesions of the spine: a rare diagnosis of eosinophilic granuloma in an adult: a case report. AB - Eosinophilic granuloma (EG) is a rare benign osteolytic lesion observed rarely in adults, with only some 18 cases of spinal location reported in the literature. We present an unusual variant of EG in a 23-year-old man with radiological features of multiple spinal lytic lesions which was evocated of metastatic processes. A surgically transpedicular biopsy of the thoracic collapsed vertebrae with posterior stabilization was made. Histological examination of the tissue showed features of eosinophilic granuloma. The clinical and radiological findings of EG present dilemmas of both diagnosis and treatment. The etiology is unclear and the therapeutic approach is still controversial. PMID- 19551335 TI - Early activation of matrix metalloproteinase-9 is associated with blood-brain barrier disruption after photothrombotic cerebral ischemia in rats. AB - BACKGROUND: The activation of matrix metalloproteinases (MMPs) is a critical event for disruption of the blood-brain barrier (BBB) during cerebral ischemia. Among the MMPs, MMP-2, and MMP-9 expression were reported to be significantly elevated after the onset of ischemia. The aim of this study was to investigate which one is more significant for BBB disruption in the photothrombotic cerebral ischemia. MATERIALS AND METHODS: Male Sprague-Dawley rats weighing 250-300 g received focal cerebral ischemia by photothrombosis. MMP-2 and MMP-9 activities were assessed by gelatin zymography at various times from 2 h to 7 days. The BBB integrity was assessed using Evans blue dye with a spectrophotometric assay. FINDINGS: The Evans blue extravasation was increased within 2 h after cerebral ischemia, and was maximal at 12 and 24 h after the injury, and then gradually decreased. MMP-9 protein activity was detected as early as 2 h after the focal ischemic event; it rapidly increased at 6 h after ischemia, and reached a maximum level 48 h after the ischemic event. Thereafter, the MMP-9 level abruptly decreased and returned to the baseline at 72 h after the insult. By contrast, the MMP-2 protein activity was up-regulated at 6 h after the focal ischemic insult, and reached a maximum level at 72 h after the event. The elevated MMP-2 levels persisted for 7 days after the injury. CONCLUSIONS: The early activation of MMP-9 was correlated with the increase in the permeability of the BBB. Our findings suggest that MMP-9 is the key factor involved in BBB disruption and subsequent brain injury after photothrombotic cerebral ischemia in rats. PMID- 19551336 TI - Association of ApoE genotype with clinical features and outcome in idiopathic normal pressure hydrocephalus (iNPH): a preliminary report. AB - Fifteen patients suspected of having iNPH were clinically evaluated and their ApoE genotype determined prior to a possible ventriculoperitoneal shunt insertion. All patients fulfilling our criteria for intervention and who had a shunt implanted with good results were homozygous for the ApoE3/3 genotype. PMID- 19551337 TI - Endonasal approaches to the sellar and parasellar regions: closure techniques using biomaterials. AB - PURPOSE: We reviewed the clinical outcomes resulting from various closure techniques used following endoscopic endonasal surgery for lesions in the sellar and parasellar regions. We compared our current closure technique, which uses a biological matrix of native equine collagen (TissuDura) fixed with fibrin sealant (Tisseel), with the technique we employed previously, using autologous materials, in order to assess the comparative efficacy and tolerability of both methods over the medium- to long-term. METHODS: A review was conducted of all cases of endonasal endoscopic intervention carried out in our institution between 1997 and 2007. Operations performed between January 1st 1997 and December 31st 2003 involved a sellar closure technique using autologous materials, either alone or supported by fibrin sealant. From January 1st 2004, sellar reconstruction techniques involving resorbable heterologous materials were used in the closure phases. Post-operatively, clinico-endoscopic assessments took place at 15 days, 1, 3, and 6 months and yearly thereafter, supplemented by magnetic resonance imaging (MRI) scanning at 3 months and annually. RESULTS: Between January 1st 1997 and December 31st 2003, 79 operations were performed in which the sellar closure technique involved the use of autologous materials. Between January 1st 2004 and January 1st 2008, 125 operations were performed in which biomaterials were used for sellar closure. The incidence of complications (fluid fistula) was 2.5% in the autologous materials closure group and 1.6% in the biomaterials closure group. The most marked difference between the two approaches was seen at 1-month follow-up, when restoration of mucociliary transport in the sphenoidal sinus and physiological functionality of the nasal mucosa and paranasal sinuses were observed to be superior in the biomaterials patient cohort. CONCLUSIONS: The development of biomaterials for closure of the sellar floor offers a viable alternative to traditional techniques using autologous materials. PMID- 19551338 TI - Clinical study on cognitive dysfunction after spontaneous subarachnoid haemorrhage: patient profiles and relationship to cholinergic dysfunction. AB - PURPOSE: We aimed to explore the cognitive profiles of subarachnoid haemorrhage patients who returned to the community, along with the associated risk factors. METHODS: We recruited 40 Chinese patients with spontaneous subarachnoid haemorrhage 7-27 months after the initial presentation. They had all been discharged to their homes or to care homes for the elderly. For cognitive assessment, we employed the Cognitive Subscale of the Alzheimer Disease Assessment Scale (ADAS-cog) for global cognitive function, the Frontal Assessment Battery (FAB) for frontal lobe function, and the Rivermead Behavioural Memory Test (RBMT) for everyday memory function. RESULTS: An ADAS-cog of more than 21/85 (poor global cognitive function) was noted in 14 (35%) patients. A FAB of less than 12/18 (poor frontal lobe function) was noted in 13 (27.5%) patients. An RBMT score of less than 15/26 (poor everyday memory function) was noted in 17 (43.6%) patients. Poor cognitive function was found to be associated with chronic hydrocephalus (in terms of FAB), with clinical vasospasm (in terms of RBMT), and with cerebral infarction (in terms of RBMT). CONCLUSIONS: Poor cognitive function was common and occurred in up to 43.6% of the patients, with the verbal and behavioural memory aspects predominantly affected. We did not find a significant association between cholinergic dysfunction and cognitive dysfunction. Organization of future drug trials and cognitive rehabilitation should take into account the association between frontal lobe dysfunction and chronic hydrocephalus. PMID- 19551339 TI - Metastasis development at the site of cervical spine arthrodesis. AB - Biologically, the site of arthrodesis provides a microenvironment replete of growth factors and active remodeling, which is propitious for bone regrowth. There is a theoretical possibility though that this microenvironment would also provide a fertile site for metastatic deposits to occur. Although spinal fusion with various types of instrumentation is commonly used to treat various disorders, development of tumors at the site of spinal arthrodesis has not been previously reported. We present the clinical, radiographic, and pathological features of a case of previously undiagnosed metastatic adenocarcinoma, occurring 1 year following anterior arthrodesis and fusion for cervical spondylotic myelopathy at the C3-C4 level. The patient's initial radiologic presentation suggested an epidural abscess with osteomyelitic destruction of C3 and C4. Our report alerts surgeons to the possibility that metastasis may occur at the site of a previous spinal arthrodesis. PMID- 19551340 TI - Ischemia of the lateral femoral cutaneous nerve during periacetabular osteotomy using Smith-Petersen approach. AB - BACKGROUND: Lateral femoral cutaneous nerve (LFCN) injury is a common complication in the Smith-Petersen approach to the hip. This complication may be induced by neural ischemia or direct trauma during the procedure. The purpose of this study was to investigate the relationship between the neural ischemia of LFCN and postoperative sensory disturbance. MATERIALS AND METHODS: Nineteen patients who underwent periacetabular osteotomy through the Smith-Petersen approach were investigated. To evaluate neural ischemia, we measured the blood flow of LFCN using a laser Doppler flowmetry. The measurements were performed before and after osteotomy at the point 1 cm distal from the lower border of the inguinal ligament. LFCN was retracted to the medial side during the procedure. There was no direct trauma to LFCN in all cases. Postoperative sensory disturbance was evaluated at 2 weeks, 3 months, and 1 year follow-up after surgery. RESULTS: After osteotomy, the blood flow of LFCN was decreased to 2.4 from 3.3 ml min(-1) 100 g(-1) when compared with that before osteotomy (P < 0.01). Postoperatively, 14 of 19 patients had sensory disturbance at 2 weeks, 8 of 19 patients at 3 months, and 2 of 19 patients at 1 year follow-up. The blood flows of both patients who had persistent symptoms over 1 year after surgery had been decreased by more than 50% during operation. CONCLUSIONS: Decrease of blood flow of LFCN by more than 50% seems to cause persistent symptoms after surgery through the Smith-Petersen approach even if direct trauma to the nerve is avoided. Excessive traction by retractors is thought to be the main cause of blood flow reduction. PMID- 19551342 TI - Analysis of interval cancers observed in an Italian mammography screening programme (2000-2006). AB - PURPOSE: The purpose of this study was to assess the interval cancer (IC) proportional incidence and review IC cases observed in an Italian mammography screening programme during 2000-2006. MATERIALS AND METHODS: ICs were identified through linkage of a screening database with the local cancer registry and hospital discharge records to calculate proportional (observed/expected) incidence. Negatively reported mammograms preceding ICs underwent blind review (randomly mixed with negative controls in a 2:1 ratio) by three expert radiologists and classified according to European guidelines criteria (OC=occult, MS=minimal sign, SE=screening error) according to majority report. Proportional IC incidence and rate of reviewed IC classified as SE were compared with European guideline standards. RESULTS: Proportional IC incidence was 10.8% in the first and 40.0% in the second year of the interval (European standard=30% or 50%, respectively). Sensitivity estimate for the 2-year interval was 74.6%. ICs were reviewed as SE, MS or OC in 15.0%, 14.0% or 71.0% of cases, respectively. Corresponding review results for negative controls were 7.0%, 25.0% or 68.0%, respectively. Positive predictive value for IC was 51.7% for SE and 21.8% for MS reporting category, respectively (p=0.008). European standard (<20% reviewed as SE) was reached. CONCLUSIONS: The study shows that the sensitivity of the mammography programme was good, complying with European guideline recommendations. Assessment of IC-based early indicators of screening efficacy is feasible in a current screening programme and should become a routine procedure. PMID- 19551341 TI - Chronic total coronary occlusion in patients with intermediate viability: value of low-dose dobutamine and contrast-enhanced 3-T MRI in predicting functional recovery in patients undergoing percutaneous revascularisation with drug-eluting stent. AB - PURPOSE: Myocardial viability was evaluated by magnetic resonance imaging (MRI) in patients with chronic total coronary occlusion (CTO) treated with a drug eluting stent. Change in left ventricular ejection fraction (LVEF) was analysed. MATERIALS AND METHODS: Twenty-three patients with CTO underwent delayed enhancement (DE) and low-dose dobutamine MRI (LD). Diastolic wall thickness (DWT), dobutamine-induced systolic wall thickening (SWT) and DE transmural extension were quantitatively assessed in vessel-related segments, calculating the contribution of viable tissue to SWT, expressed as viability index (VI)=[SWTx(100 - DE)]/100. Patients with transmural enhancement were excluded from revascularisation. At 6 months follow-up, patients underwent coronary angiography (CA) and MRI. Functional recovery was defined as a 2-mm increase in SWT. RESULTS: Transmural enhancement (mean DE 62.88+/-37.18] was present in three patients. Mean DWT, SWT, VI and DE of recanalised patients were 8.03+/-2.35, 2.64+/-1.56, 1.77+/-1.48 mm and 41.97+/-30.32. Revascularisation was successful in 14/16. Follow-up CA showed patency of treated vessels. Functional recovery was achieved in 13 patients. Functional recovery showed significant correlation with SWT (beta 1,779, p=0.015), and even higher correlation with VI (beta 2.032, p=0.011). LVEF improved significantly [Delta 95% confidence interval (CI) -4.47, p=0.0203). CONCLUSIONS: Invasive CTO treatment has beneficial effects on myocardial contractility that can be predicted by VI, and on LVEF. PMID- 19551343 TI - Assessment of left ventricular volumes with cardiac MRI: comparison between two semiautomated quantitative software packages. AB - PURPOSE: This study compared two quantitative semiautomated software packages for volumetric analysis of the left ventricle (LV) by magnetic resonance (MR) imaging using two-dimensional (2D) cine balanced steady-state free precession (b-SSFP) sequences. MATERIALS AND METHODS: We included 46 consecutive patients who underwent cardiac MR imaging for various indications. Two-dimensional cine b-SSFP sequences were used to assess the LV. Data sets were evaluated with two dedicated software packages: ViewForum, version 4.2, and Argus, version Va60C. Results were compared with Student's t test for paired samples, Pearson's r correlation coefficient and R2 coefficient of determination; ejection fraction differences were assessed with Bland-Altman analysis. The time required for analysis was also recorded. RESULTS: We observed very high levels of concordance and reproducibility. High correlation was observed for ejection fraction (p>0.05; r=0.9; R (2)=0.82). The time required for analysis was 7.6+/-2.78 min vs. 7.52+/ 2.4 min (p>0.05; r=0.85; R (2)=0.73). Intraobserver and interobserver variability did not show significant differences. CONCLUSIONS: LV volume evaluation is an integral part of cardiac MR imaging. In our experience, there is no significant difference between the commonly used software packages in either quantitative output or time required for analysis. PMID- 19551344 TI - Prognostic value of multidetector computed tomography in bowel infarction. AB - PURPOSE: This study aimed to correlate computed tomography (CT) findings and outcomes in patients affected by bowel infarction. MATERIALS AND METHODS: Twenty seven patients with bowel infarction due to vascular obstruction were evaluated with multidetector CT (MDCT) to establish the prognostic value of CT findings and their correlation with the origin of the ischaemia. The chi-square test was used to analyse the results (por= 20%) and one minor QTL (FM.2.1; R (2) = 8%) were found to be associated with FM. This map was then merged with a previous recombinant inbred line (RIL)-based map used to identify seven QTL associated with QbetaC in melon fruit. This consensus map [300 molecular markers (187 co-dominant melon and 14 interspecific; 10 LG)] provides a framework for the further dissection and cloning of published QTL, which will consequently lead to more effective trait introgression in melon. PMID- 19551369 TI - Circulating angiopoietin-2 levels in the course of septic shock: relation with fluid balance, pulmonary dysfunction and mortality. AB - PURPOSE: To investigate whether angiopoietin-2, von Willebrand factor (VWF) and angiopoietin-1 relate to surrogate indicators of vascular permeability, pulmonary dysfunction and intensive care unit (ICU) mortality throughout the course of septic shock. METHODS: In 50 consecutive mechanically ventilated septic shock patients, plasma angiopoietin-2, VWF and angiopoietin-1 levels and fluid balance, partial pressure of oxygen/inspiratory oxygen fraction and the oxygenation index as indicators of vascular permeability and pulmonary dysfunction, respectively, were measured until day 28. RESULTS: Angiopoietin-2 positively related to the fluid balance and pulmonary dysfunction, was higher in non-survivors than in survivors and independently predicted non-survival throughout the course of septic shock. VWF inversely related to the fluid balance and pulmonary dysfunction throughout the course of septic shock, was comparable between survivors and non-survivors and predicted non-survival on day 0 only. Angiopoietin-1 positively related to pulmonary dysfunction throughout the course, but did not differ between survivors and non-survivors. CONCLUSIONS: In contrast to VWF, plasma angiopoietin-2 positively relates to fluid balance, pulmonary dysfunction and mortality throughout the course of septic shock, in line with a suggested mediator role of the protein. PMID- 19551370 TI - Decontamination of the digestive tract and oropharynx: hospital acquired infections after discharge from the intensive care unit. AB - OBJECTIVE: To determine the incidence rates of hospital acquired infections (HAI) during the first 14 days after ICU discharge after treatment during ICU-stay with Selective Decontamination of the Digestive tract (SDD), Selective Oropharyngeal Decontamination (SOD) or Standard Care (SC). DESIGN: Prospective observational study. SETTING: ICUs in two tertiary care hospitals. PATIENTS: Patients discharged from the ICU to the ward. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Post-ICU incidences of HAI per 1,000 days at risk were 11.2, 12.9 and 8.3 for patients that had received SDD (n = 296), SOD (n = 286) or SC (n = 289) respectively in ICU, yielding relative risks, as compared to SC, of 1.49 (CI(95) 0.9-2.47) for SOD and 1.44 (CI(95) 0.87-2.39) for SDD. Incidences of surgical site infections (per 100 surgical procedures) were 4 after SC and 11.8 and 8 after SOD and SDD (p = 0.04). Among patients that succumbed in the hospital after ICU-stay (n = 58) eight (14%) had developed HAI after ICU discharge; 3 of 21 after SDD, 3 of 15 after SOD and 2 of 22 after SC. CONCLUSIONS: Incidences of HAI in general wards tended to be higher in patients that had received either SDD or SOD during ICU-stay, but it seems unlikely that these infections have an effect on hospital mortality rates. PMID- 19551371 TI - Post-resuscitation NOS inhibition does not improve hemodynamic recovery of hypoxic newborn pigs. AB - BACKGROUND: Significant improvement in myocardial recovery has been shown previously with interventions to decrease reactive oxygen species after ischemia/hypoxia. We investigated whether co-administration of N-acetylcysteine (NAC, a scavenger for reactive oxygen species) and N (G)-monomethyl-L: -arginine (L-NMMA, a non-selective nitric oxide synthase inhibitor) results in better hemodynamic recovery. DESIGN: Controlled, block-randomized study. SETTING: University research laboratory. SUBJECT: Mixed breed piglets (1-4d, 1.6-2.4 kg). INTERVENTIONS: Acutely instrumented piglets received normocapnic alveolar hypoxia (10-15% oxygen) for 2 h followed by reoxygenation with 100% oxygen (1 h) then 21% oxygen (3 h). After reoxygenation, hypoxic-reoxygenated piglets were given either saline (controls), NAC [30 mg/kg bolus + 20 mg/(kg h) infusion], NMMA [0.1 mg/kg bolus + 0.1 mg/(kg h) infusion] or NAC + L-NMMA via intravenous infusion in a blinded, randomized fashion (n = 8/group). Sham-operated piglets had no hypoxia reoxygenation (n = 5). MEASUREMENTS AND RESULTS: Both cardiac index and stroke volume of hypoxia-reoxygenation controls remained depressed during reoxygenation (vs. normoxic baseline, p < 0.05). Post-resuscitation treatment with L-NMMA alone did not improve systemic hemodynamic recovery, but caused pulmonary hypertension (vs. controls). In contrast, treating the piglets with either NAC or NAC + L-NMMA improved cardiac index and stroke volume, with no effect on heart rate and blood pressure (vs. controls). These treatments also decreased various oxidative stress markers in myocardial tissues (vs. controls). However, there was no significant difference between NAC- and NAC + L-NMMA groups in all examined parameters. CONCLUSIONS: Post-resuscitation administration of NAC improved cardiac function and reduced oxidative stress in newborn pigs with hypoxia-reoxygenation insult. Low-dose, non-selective inhibitor of nitric oxide synthase activity did not provide any further beneficial effect. PMID- 19551373 TI - Lack of sedative effects after vespertine intake of oxazepam as hypnotic in healthy volunteers. AB - AIMS: An objective physiological test was used to investigate the hangover effect, its time course and dose relationship compared to placebo and an herbal relaxant. METHODS: Pupillographic Sleepiness Test as an objective measurement, Stanford Sleepiness Scale (SSS) and visual analogue scales (VAS) were used. Study design included: (a) randomised, double-blind, double-dummy, placebo-controlled crossover trial; (b) double-blind, placebo-controlled, randomised study. Primary end point was the Pupillary Unrest Index (lnPUI). RESULTS: Oxazepam 10 mg did not increase PUI. In the VAS and SSS, there was no increase in sleepiness after the three treatment periods. Neither 10 nor 30 mg oxazepam caused sedation in healthy volunteers. Subjective and objective sleepiness measures correlated significantly. DISCUSSION: The lack of sedative effects after vespertine intake of oxazepam (10/30 mg) seems to be relevant with respect to product safety. With regard to the subjective perception at 30 mg, fatigue rather than sleepiness may be the underlying reason. PMID- 19551372 TI - Patients' preferences for enrolment into critical-care trials. AB - BACKGROUND: Most critically ill patients are incapable of providing informed consent for research. OBJECTIVE: We sought to determine patients' preferences for different consent frameworks for enrolling incapable patients into critical-care trials. DESIGN: Prospective observational and structured interview study. SETTING: Five university-affiliated hospitals in Ontario. PATIENTS: Two-hundred and forty consecutive capable and consenting survivors of critical illness. INTERVENTION: Participants considered four frameworks for enrolling incapable patients into clinical trials using a baseline scenario and three permutations for: risk (very low vs. high), treatment type (new vs. currently available), and availability of substitute decision-maker (yes vs. no). MEASUREMENTS AND MAIN RESULTS: For each scenario, patients chose their preferred framework and rated the acceptability of each framework using a seven-point Likert scale. Most (180/240; 76%) patients selected "consent by substitute prior to enrolment" as their preferred framework; this also received the highest baseline acceptability ratings ("acceptable" or "highly acceptable" 207/240; 87%). Modifying risk or treatment type did not substantially change these ratings. A minority of patients rated delayed consent as unacceptable or highly unacceptable in both the baseline scenario (48/240, 20% delayed to substitute; 57/240, 24% delayed to patient) and when a substitute was unavailable (34/240; 15%). CONCLUSIONS: Most survivors of critical illness found the usual practice of obtaining informed consent from a substitute decision-maker prior to enrolment in a clinical trial to be acceptable. Nearly half of patients considered foregoing informed consent to be unacceptable, whereas a minority considered enrolment followed by delayed consent to be unacceptable even when a substitute was unavailable. These approaches should, therefore, only be considered when deviating from the usual practice of obtaining consent from a substitute decision-maker is truly justified, such as where treatments being tested need to be delivered as soon as possible in order to be effective. PMID- 19551374 TI - Traceability of sulfonamide antibiotic treatment by immunochemical analysis of farm animal hair samples. AB - The use of hair to trace use of unauthorized substances, therapeutic agents, or their misuse is becoming very attractive since residues can be detected for a long time after treatment. For this purpose, an indirect enzyme-linked immunosorbent assay (ELISA) has been evaluated for its capability to trace sulfonamide antibiotic treatment by analyzing cattle and pig hair samples. Pigmented and nonpigmented hair samples from control and sulfamethazine (SMZ) treated pigs and calves were collected, extracted under different alkaline conditions, and analyzed by ELISA after just diluting the extracts with the assay buffer. Data analysis following the European recommendations for screening methods demonstrates that the ELISA can detect SMZ in hair samples with a limit of detection (90% of the zero dose (IC(90))) between 30 and 75 ng g(-1). The same samples have been analyzed by HPLC after a dual solid-phase extraction. The ELISA results matched very well those obtained by the chromatographic method, demonstrating that the immunochemical method can be used as a screening tool to trace animal treatments. Between the benefits of this method are the possibility to directly analyze hair extracts with sufficient detectability and its high throughput capability. Preliminary validation data are reported using an experimental approach inspired on the Commission Decision 2002/657/EC criteria for screening methods. PMID- 19551375 TI - Can illusory deviant stimuli be used as attentional distractors to record vMMN in a passive three stimulus oddball paradigm? AB - A passive three stimulus oddball paradigm was used to investigate Visual Mismatch Negativity (vMMN) a component of the Event Related Potential (ERP) believed to represent a central pre-attentive change mechanism. Responses to a change in orientation were recorded to monochrome stimuli presented to subjects on a computer screen. One of the infrequent stimuli formed an illusory figure (Kanizsa Square) aimed to capture spatial attention in the absence of an active task. Nineteen electrodes (10-20 system) were used to record the electroencephalogram in fourteen subjects (ten females) mean age 34.5 years. ERPs to all stimuli consisted of a positive negative positive complex recorded maximally over lateral occipital areas. The negative component was greater for deviant and illusory deviant compared to standard stimuli in a time window of 170-190 ms. A P3a component over frontal/central electrodes to the illusory deviant but not to the deviant stimulus suggests the illusory figure was able to capture attention and orientate subjects to the recording. Subtraction waveforms revealed visual discrimination responses at occipital electrodes, which may represent vMMN. In a control study with 13 subjects (11 females; mean age 29.23 years), using an embedded active attention task, we confirmed the existence of an earlier (150-170 ms) and attenuated vMMN. Recordings from an intracranial case study confirmed separation of N1 and discrimination components to posterior and anterior occipital areas, respectively. We conclude that although the illusory figure captured spatial attention in its own right it did not draw sufficient attentional resources from the standard-deviant comparison as revealed when using a concurrent active task. PMID- 19551376 TI - Differential expression of two glutamate transporters, GLAST and GLT-1, in an experimental rat model of glaucoma. AB - Glutamate is the major excitatory neurotransmitter of the mammalian retina, and excessive glutamate has been implicated in the pathogenesis of glaucoma. It is well known that glutamate transport, mainly via GLAST and GLT-1, is cardinal mechanism for maintaining glutamate homeostasis in normal and pathological conditions, including ischemia in the brain. In an effort to understand the role of glutamate and the glutamate regulation system of the retina in the pathogenesis of glaucoma, we examined changes in the expression of two glutamate transporters, GLAST and GLT-1, by Western blot analysis and immunocytochemistry in a rat glaucoma model. GLT-1 was expressed in cone photoreceptors and some cone bipolar cells and the levels of expression were significantly increased in the cauterized eyes throughout the entire experimental period. In contrast, GLAST expression, which occurred in Muller cells, the main retinal glial cells, remained stable during the experimental period. These results suggest that GLT-1 may be a prerequisite for the maintenance of glutamate homeostasis in the retina undergoing glaucoma. PMID- 19551377 TI - Challenges in quantifying multisensory integration: alternative criteria, models, and inverse effectiveness. AB - Single-neuron studies provide a foundation for understanding many facets of multisensory integration. These studies have used a variety of criteria for identifying and quantifying multisensory integration. While a number of techniques have been used, an explicit discussion of the assumptions, criteria, and analytical methods traditionally used to define the principles of multisensory integration is lacking. This was not problematic when the field was small, but with rapid growth a number of alternative techniques and models have been introduced, each with its own criteria and sets of implicit assumptions to define and characterize what is thought to be the same phenomenon. The potential for misconception prompted this reexamination of traditional approaches in order to clarify their underlying assumptions and analytic techniques. The objective here is to review and discuss traditional quantitative methods advanced in the study of single-neuron physiology in order to appreciate the process of multisensory integration and its impact. PMID- 19551379 TI - Ultrasound of the hands and feet for rheumatological disorders: influence on clinical diagnostic confidence and patient management. AB - PURPOSE: The purpose of the study was to quantify the impact that ultrasound (US) of the hands and feet has on the rheumatologists' diagnostic confidence and on patient management. MATERIALS AND METHODS: There were 62 consecutive referrals from two rheumatologists for US of the hands and/or feet for this prospective controlled observational study. Measurements of diagnostic confidence for both specific clinical findings as well as overall diagnosis using a Likert scale were made both before and after the US examination in each case. Proposed management was also recorded before US and then with the benefit of the US result. McNemar's test was performed to determine differences in diagnostic certainty and proposed management before and after US. RESULTS: The physician certainty for specific clinical findings increased significantly following US for synovitis (9.7 vs 38.7%), tenosynovitis (9.7 vs 46.8%), erosions (1.6 vs 58.1%), enthesitis (50.0 vs 83.9%) and other (53.2 vs 77.4%). The physician certainty for overall diagnosis increased significantly for seronegative arthritis (46.8 vs 61.3%), inflammatory osteoarthritis (46.8 vs 87.1%), and primary osteoarthritis (46.8 vs 73.0%). A total of 88.7% of patients had disease-modifying antirheumatic drugs as a proposed management option before US vs 48.4% after US. Before US, 4.8% of patients had non-steroidal anti-inflammatory drug as a proposed management option versus 45.2% after US. CONCLUSION: Ultrasound of the hands and/or feet significantly influenced the rheumatologists' diagnostic confidence in specific clinical findings and management plans. PMID- 19551380 TI - Perineal nodular swelling in a recreational cyclist: diagnosis and discussion. PMID- 19551378 TI - Molecular identification and dynamics of microbial communities in reactor treating organic household waste. AB - The prokaryotic diversity associated with organic household waste (OHW), leachate (start-up inoculum), and mesophilic anaerobic digestion processes in the degradation of OHW for 44 and 90 days was investigated using a culture independent approach. Bacterial and archaeal 16S rRNA and mcrA gene clone libraries were constructed from community DNA preparations. Bacterial clones were affiliated with 13 phyla, of which Firmicutes, Proteobacteria, and Bacteroidetes were represented in all libraries, whereas Actinobacteria, Thermotogae, Lentisphaerae, Acidobacteria, Chloroflexi, Cyanobacteria, Synergistetes, Spirochaetes, Deferribacteres, and Deinococcus-Thermus were exclusively identified in a single library. Within the Archaea domain, the Euryarchaeota phylum was the only one represented. Corresponding sequences were associated with the following orders of hydrogenotrophic methanogens: Methanomicrobiales (Methanoculleus genus) and Methanobacteriales (Methanosphaera and Methanobacterium genera). One archaeal clone was not affiliated with any order and may represent a novel taxon. Diversity indices showed greater diversity of Bacteria when compared to methanogenic Archaea. PMID- 19551381 TI - Treatment combinations targeting apoptosis to improve immunotherapy of melanoma. AB - Immunotherapy based on T cell responses to the tumor is believed to involve killing of cancer cells by induction of apoptosis. The predominant mechanisms are death ligand-induced signaling mainly by TNF-related apoptosis-inducing ligand (TRAIL) mediated by CD4 T cells, monocytes and dendritic cells, and perforin/granzyme mediated apoptosis mediated by CD8 T cells and NK cells. Resistance against TRAIL involves loss of TRAIL death receptors and/or activation of the MEK and/or Akt signal pathways. Resistance to CD8 CTL responses also involves activation of the MEK and/or Akt pathways. Apoptosis induced by immune responses is regulated by the Bcl-2 family of proteins. Many reagents have been developed against the Bcl-2 antiapoptotic proteins and clinical trials combining them with immunotherapy are awaited. The second group of agents that regulate the Bcl-2 family of proteins are the signal pathway inhibitors. Clinical trials with inhibitors of RAS, RAF or MEK are in progress and would appear an exciting combination with immunotherapy. One of the main drivers of resistance to apoptosis are adaptive mechanisms that allow cancer cells to overcome endoplasmic reticulum (ER) stress. These adaptive mechanisms inhibit practically all known apoptotic pathways and create an acidic environment that may reduce infiltration of lymphocytes against the tumor. The signal pathway inhibitors may be effective against these adaptive processes but additional agents that target ER stress pathways are in development. In conclusion, combination of immunotherapy with agents that target antiapoptotic mechanisms in cancer cells offers a new approach that requires evaluation in clinical trials. PMID- 19551382 TI - Phase I/II and pharmacokinetic study of S-1 and oxaliplatin in previously untreated advanced gastric cancer. AB - BACKGROUND: We aimed to determine the maximum-tolerated dose (MTD) of S-1 when given with oxaliplatin, to evaluate S-1 pharmacokinetics, and to determine the efficacy and safety of this regimen as a first-line treatment for advanced gastric cancer (AGC). METHODS: Oxaliplatin was fixed at a dose of 130 mg/m2 on day 1 (D1). S-1 was administered from D1 to D14 of a 3-week cycle, and escalated by 10 mg/m2 per day from 70 mg/m2 per day up to 100 mg/m2 per day. Pharmacokinetic analyses were performed following a single dose of S-1 on D-5 and D1 of the first cycle. RESULTS: In phase I (n=18), MTD was not defined. In phase II (n=47) with the planned maximum dose, partial response was achieved in 26 patients (55.3%) and stable disease in 14 patients (29.8%). The median time to progression was 6.6 months (95% CI 4.0-9.2 months) and the median overall survival was 12.5 months (95% CI 9.2-15.9 months). Frequent grade 3/4 toxicities included thrombocytopenia (39%), neutropenia (28%), anemia (17%), and leukopenia (13%). There was one grade 5 febrile neutropenia during the first cycle. CONCLUSIONS: The pharmacokinetics of S-1 was not influenced by oxaliplatin. S 1/Oxaliplatin combination therapy is highly active against AGC and has a favorable toxicity profile. PMID- 19551383 TI - SAPHO syndrome in an adult with ulcerative colitis responsive to intravenous pamidronate: a case report and review of the literature. AB - Synovitis, acne, pustulosis, hyperostosis, osteitis (SAPHO) syndrome is a rare group of sterile, inflammatory osteoarticular disorders classically associated with skin lesions. It is occasionally associated with enteropathic disease such as ulcerative colitis. We present a 39-year-old patient with chronic ulcerative colitis who developed enteropathic SAPHO and responded well to pamidronate. We discuss the clinicopathological features with particular attention to bone pathology, and perform a literature review of this fascinating syndrome. PMID- 19551384 TI - Successful resolution of pneumonia developed in a patient affected by Crohn's disease, rheumatoid arthritis, myasthenia gravis and recurrent uveitis during concomitant treatment with tumour necrosis factor alpha inhibitors and conventional immunosuppressive drugs. AB - Tumour necrosis factor alpha inhibitors, both infliximab and adalimumab, have been approved for the treatment of both rheumatoid arthritis and Crohn's disease. A slight increase in the risk of infections in patients receiving immunosuppressants and/or biological agents has been reported. Here, we present the case of a 68-year-old woman affected by Crohn's disease, myasthenia gravis, recurrent uveitis and rheumatoid arthritis who developed pneumonia during concomitant treatment with biological agents and conventional immunosuppressive drugs. PMID- 19551386 TI - Bladder pain syndrome/interstitial cystitis: a sense of urgency. AB - A classic triad of symptoms (bladder pain, urinary frequency, and urgency) has served to define bladder pain syndrome/painful bladder syndrome/interstitial cystitis (BPS/PBS/IC) syndrome. BPS/PBS/IC is a distinct condition and it is likely that the urgency experienced by these patients differs from that experienced by those with overactive bladder syndrome. It is unclear how best to define urgency in the BPS/PBS/IC setting. Differences in the other primary symptoms associated with these conditions probably influence how urgency is perceived. Advances in research into the pathophysiology of urgency and underlying disease processes will help to optimize both the diagnosis and treatment of BPS/PBS/IC. PMID- 19551385 TI - Stress-related variation in antioxidative enzymes activity and cell metabolism efficiency associated with embryogenesis induction in isolated microspore culture of triticale (x Triticosecale Wittm.). AB - Isolated microspore cultures of two spring triticale (x Triticosecale Wittm.) cultivars were used to examine the effect of various stress treatments (either high--32 degrees C or low--5 degrees C temperature with or without nitrogen/carbohydrate starvation) applied to excised anthers on the effectiveness of microspore embryogenesis induction. To quantify the effects of pretreatment conditions, the activity of antioxidative enzymes (catalase, peroxidase and superoxide dismutase) together with respiration rate and heat emission were measured. It was observed that heat shock treatment applied as the only one stress factor increased the activity of antioxidative enzymes which suggests intensive generation of reactive oxygen species. Such pretreatment effectively triggered microspore reprogramming but drastically decreased microspore viability. After low temperature treatment, the activity of antioxidative enzymes was similar to the control subjected only with the stress originated from the transfer to in vitro culture conditions. This pretreatment decreased the number of microspores entering embryogenesis but sustained cell viability and this effect prevailed in the final estimation of microspore embryogenesis effectiveness. For both, low- and high-temperature treatments, interaction with starvation stress was beneficial increasing microspore viability (at 5 degrees C) or efficiency of embryogenesis induction (at 32 degrees C). The latter treatment significantly reduced cell metabolic activity. Physiological background of these effects seems to be different and some hypothetical explanations have been discussed. Received data indicate that in triticale, anther preculture conditions could generate oxidative stress and change the cell metabolic activity which could next be reflected in the cell viability and the efficiency of microspore embryogenesis. PMID- 19551387 TI - Intracranial tumors in first year of life: the CHEO experience. AB - INTRODUCTION: One seventh of pediatric brain tumors are diagnosed in the first year of life. With more widespread and accessible neuroimaging, these lesions are being diagnosed earlier, but there remains scant literature about their natural history. METHODS: A retrospective review was performed of brain tumor patients presenting to the Children's Hospital of Eastern Ontario (CHEO) through the last 34 years. Patients presenting in the first year of life, including symptoms, management features, and functional outcome, were analyzed using ANOVA and chi (2) statistics. RESULTS: Eighteen cases of brain tumors in the first year of life were identified: 12 suptratentorial, eight with benign histology, and six infratentorial all with malignant histology. Median age of presentation differed by lesion location (p = 0.05) and glial tumors were most common. Raised intracranial pressure was more than twice as prevalent with posterior fossa lesions (p < 0.01) with equivalent likelihood of increasing head circumference (p = 0.74), whereas seizures were more frequent with supratentorial tumors (p = 0.04). Gross total resection was achieved in 47% of patients, cerebrospinal fluid diversion was more frequently necessary among infratentorial lesions (p = 0.02), and adjuvant therapy was more utilized for infratentorial lesions (p < 0.01). Among eight surviving infants, seven had supratentorial tumors, five survived to adulthood, and six are functionally independent. CONCLUSIONS: Brain tumors in the first year of life represent 4.8% of patients treated at CHEO. Mode of presentation, utilization of adjuvant therapy, and survival depend on tumor location and histology, with worse prognosis for infratentorial lesions. One third of patients had acceptable functional outcome requiring no special assistance. PMID- 19551388 TI - Cervical myelocystocele: prenatal diagnosis and therapeutical considerations. PMID- 19551389 TI - Defenestration in children younger than 6 years old: mortality predictors in severe head trauma. AB - PURPOSE: This study aims to describe the characteristics of severe head injuries in children less than 6 years old, victims of falls from windows, and identify the main predictive factors of mortality in this population. PATIENTS AND METHODS: A cross-sectional study was designed through data derived from medical records of less than 6-year-old children victims of falls from windows presenting with a severe head injury defined by an initial Glasgow coma scale (GCS) < or =8, hospitalized at a Pediatric Trauma center level III, between January 2000 and December 2005. Statistical analysis used univariate analysis and multiple logistic regressions. RESULTS: We identified 58 severe head injuries in children victims of falls from windows. The mean age was 2.8 +/- 1.4 years, with a male prevalence (64%); 48% of patients had a GCS < or =5; 62.1% had a Pediatric Trauma Score (PTS) < or =3 at hospital admission. The mortality rate was 41% (24/58) and most of them (88%; 21/24) died within 48 h. An increased death rate was noted in children admitted with hypoxemia (p = 0.001), low systolic blood pressure (p = 0.002), hypothermia (p = 0.0001), GCS < or =5 (p = 10(-5)), PTS < or =3 (p = 0.008), hyperglycemia (p = 0.023), coagulation disorders (p = 0.02), and initial intracranial pressure > or =20 mmHg (p = 0.03). Initial hypothermia, hyperglycemia, and coagulation disorders were the only independent predictive factors of mortality. CONCLUSION: Severe head injuries resulting from falls from windows carry a high risk of mortality in less than 6-year-old children. Hypothermia, hyperglycemia, and coagulation's disorders are independent predictive factors of mortality. Early deaths could be considered as direct consequences of uncontrollable brain lesions. PMID- 19551390 TI - Bilateral choroid plexus hyperplasia: a case report and management strategies. AB - INTRODUCTION: Choroid plexus hyperplasia (CPH) is a rare cause of cerebrospinal fluid (CSF) overproduction and shunt-resistant hydrocephalus in infants. If treated with a ventriculoperitoneal (VP) shunt, these patients secondarily develop CSF accumulation along the shunt tract and within the peritoneum. The surgical management of this condition is not as clearly defined as in the case of a choroid plexus papilloma or carcinoma. CASE REPORT: An 8-day-old male patient presented with bulging fontanelle, head circumference = 42 cm (>98th percentile), and cranial ultrasound demonstrating communicating hydrocephalus with enlarged choroid plexuses. A VP shunt was inserted, and secondarily progressive hydrocephalus, shunt tract fluid accumulation, and ascites developed. The infant underwent staged bilateral plexectomy and is tolerating CSF diversion. The pathology confirmed CPH. DISCUSSION: Of the cases reported in the literature, treatments have included CSF shunting, endoscopic coagulation, and craniotomy with plexectomy. CSF shunting was required in the majority but not all. Only those having undergone bilateral choroid plexectomy have been rendered shunt free. PMID- 19551391 TI - Duodenal atresia: associated anomalies, prenatal diagnosis and outcome. AB - BACKGROUND: The diagnosis of duodenal atresia is commonly made prenatally, either as an isolated lesion or due to its association with other chromosomal abnormalities (Robertson et al. in Semin Perinatol 18:182-195, 1994; Hemming and Rankin in J Prenat Diagn 27:1205-1211, 2007). The aim of this study was to describe the prevalence of associated anomalies, prenatal diagnostic accuracy and survival of cases of congenital duodenal atresia in our institution. METHODS: All cases of duodenal atresia registered with our local congenital anomaly register over a 10-year period, 1995-2004 inclusive, were studied, including those resulting in termination of pregnancies, stillbirths, intrauterine deaths and neonatal deaths. To ensure high-case ascertainment, data were cross checked with prenatal ultrasound, cytogenetic laboratory, pathology department and neonatal surgical data base. Data were analysed for associated anomalies, accuracy of prenatal diagnosis and neonatal outcomes. RESULTS: A total of 65 patients were initially diagnosed as having duodenal atresia, of these 4 were subsequently excluded (1 postnatal normal bowel and 3 high jejunal atresias). In the remaining 61 cases, 35 (57%) had an association with other congenital abnormalities and 26 (43%) were isolated anomalies. Thirty-five were male and 26 female (M:F = 1.4:1). Twenty-one out of 29 (72%) patients prenatally diagnosed, compared with 14 out of 32 (44%) patients diagnosed postnatally had associated anomalies. Duodenal atresia was suspected on routine prenatal ultrasonography at 20-week gestation in 33 cases and confirmed in 29 (48%) cases with 4 false-positive diagnoses (1 normal bowel and 3 high jejunal atresias). No prenatal diagnosis was made in 32 (52%) babies. Of the 61 cases, 53 were live births with 2 early neonatal deaths (1 cardiac and 1 VACTERL), 5 terminations, 2 intrauterine deaths and 1 stillbirth (Fig. 3). Overall neonatal survival was 96% (51 cases). Mortality in the group diagnosed prenatally was 34 % (10 cases). CONCLUSION: This study shows an overall increased association of duodenal atresia with Down's syndrome. In the group diagnosed prenatally, mortality as well as the association with other congenital anomalies was found to be higher. We have demonstrated a greater prenatal diagnostic accuracy, but confirm postnatal outcomes similar to previous studies. PMID- 19551392 TI - [Anti-B-cell strategies in vasculitides and collagenoses]. AB - B-cells play a central role in the pathogenesis of autoimmune diseases. As already discussed in other articles, besides the production of potentially pathogenic autoantibodies, B-cells may function as antigen-presenting cells, may induce T-cell activation and produce various cytokines. The feasibility of targeting B-cells in patients with severe and refractory autoimmune disorders, especially in patients with vasculitis or connective tissue diseases, has met growing interest among rheumatologists in recent years. The use of rituximab as a monoclonal antibody directed against CD20 positive B-cells has been reported in case reports and small patient series; however, these are hard to compare as different diseases are described and different doses and schedules of rituximab were used. It has to be considered that positive reports are more likely to be reported than patients who do not improve or experience side effects. Data on only a few indications from randomized, double-blind studies are available; however, even these results should be evaluated critically. PMID- 19551393 TI - Geographical variation in incidence of primary total hip arthroplasty: a population-based analysis of 34,642 replacements. AB - BACKGROUND: Considerable variation in total hip arthroplasty (THA) incidence between regions has been described. The aim of this study was to evaluate geographical variation in the incidence of primary THA for OA in Finland and to analyze variables associated with this variation. METHODS: Using Hospital Discharge Register, 34,642 THAs performed for primary OA over the 1998 and 2005 period were identified. Incidence indices for 21 hospital administrative regions were determined. Logistic regression analysis and generalized linear models were used for studying the association of potential explanatory factors for the variation in the incidence of THA. RESULTS: Incidence indices of THA varied from 1.9- to 3.0-fold during the study period. A high ratio of primary THA for primary OA to primary THA for any reason was associated with a high absolute incidence of primary THA (P < 0.001). Neither average incomes nor morbidity were associated with the incidence of THA. INTERPRETATION: When hip surgery was performed on a larger scale, threshold for performing THAs due to primary OA was higher. Socio economic status had no apparent effect on THA rate. Surgeon decision-making related factors influence THA rates when there are only a few surgeons responsible for performing THAs in a region. PMID- 19551394 TI - Recurrent mutations in functionally-related EDA and EDAR genes underlie X-linked isolated hypodontia and autosomal recessive hypohidrotic ectodermal dysplasia. AB - Mutations in three functionally related genes EDA, EDAR and EDARDD have been reported to cause hypohidrotic ectodermal dysplasia (HED), which is characterized by sparse hair, reduced ability to sweat, and hypodontia. In few cases mutations in the EDA gene have been found to result in X-linked recessive isolated hypodontia. In the study, presented here, we have ascertained two large Pakistani families (A and B) with autosomal recessive form of hypohidrotic ectodermal dysplasia and X-linked recessive isolated hypodontia. Genetic mapping showed linkage of family A to EDAR gene on chromosome 2q11-q13 and family B to EDA gene on chromosome Xq12-q13.1. Subsequently, DNA sequencing of the coding regions of EDAR and EDA genes revealed previously described mutations. Sequence analysis identified a four base-pair splice-junction deletion mutation (c.718_721delAAAG) in EDAR gene in family A and a missense mutation (c.T1091C; p.M364T) in EDA gene in family B. Recurrence of mutations in EDAR and EDA genes in unrelated families is evocative of the dispersion of ancestral chromosome in different locality groups through common ancestors. PMID- 19551395 TI - Right lower quadrant pain and postpartum ovarian vein thrombosis. Uncommon but not forgotten. AB - INTRODUCTION: Ovarian vein thrombosis (OVT) is an uncommon, life-threatening complication of the postpartum period. Symptoms of OVT include lower abdominal and flank pain associated with fever, tachycardia, and nausea and vomiting. CASE HISTORY: We present a case of OVT in a 22-year-old woman 4 days postpartum presenting to the emergency department with right lower quadrant pain, tachycardia, and leukocytosis. The initial clinical diagnosis was of appendicitis; however, on CT scan, she had findings characteristic of right OVT. CONCLUSION: The clinical presentation of OVT can be very similar to that of acute appendicitis and should always be included in the differential diagnosis of right lower quadrant pain in the female patient. Modern imaging techniques enable confirmation of the diagnosis when OVT is suspected. PMID- 19551396 TI - The unbooked mother: a cohort study of maternal and foetal outcomes in a North London Hospital. AB - OBJECTIVE: The aim of the study was to determine the pregnancy outcome of unbooked compared to booked women of similar parity and ethnic background over a period of 18 months at the North Middlesex University Hospital (NMUH), London. METHODS: A retrospective cohort study from September 2006 to March 2008 comparing the socio-demographics, foetal and maternal outcomes of pregnancies of unbooked versus booked women. Women who received no antenatal care or who delivered within 3 days of the initial booking visit were categorized as 'unbooked' while the next delivered women on the labour ward register (matched for ethnicity and parity) who booked prior to the second trimester served as comparison. Fischer's exact test (two-tailed), student's t test and Pearson's chi-square test, odds ratio and 95% confidence interval were used to compare the two groups statistically. RESULTS: There were 91 unbooked births in the 18 month study period. Unbooked mothers were younger (26.0 +/- 6.68 vs. 29.2 +/- 6.10 years, p < 0.0001), more likely to be unemployed, unmarried and non-English speaking compared to matched booked mothers (all p < 0.01). The rate of Caesarean sections ([OR] = 0.85, 95% [CI] = 0.39-1.88), p > 0.05), birth asphyxia, Apgar scores <7 at 1 min ([OR] = 1.10, 95% [CI] = 0.52-2.35, p = 0.80) and intrauterine foetal death (stillbirth) ([OR] = 2.03, 95% [CI] = 0.28-14.60, p = 0.60) were comparable between the two groups. However, unbooked mothers were five times more likely to have preterm delivery ([OR] = 6.44, 95% [CI] = 2.24-18.50, p < 0.0002); three times more likely to have low birth weight babies (<2,500 g) ([OR] = 2.87, 95% [CI] = 1.21 6.82, p < 0.02) and twice as likely to have postpartum haemorrhage ([OR] = 1.85, 95% [CI] = 0.69-4.98, p = 0.3). CONCLUSION: Unbooked women were more at risk of adverse foetal and maternal outcomes than booked women, even within a population of young, relatively healthy immigrant women. PMID- 19551398 TI - Comparison of the h index with standard bibliometric indicators to rank influential otolaryngologists in Europe and North America. AB - The Hirsch (h) index is an original and simple new bibliometric measure incorporating both quantity and quality. In this study, our aim was first to present characteristics of the statistical correlation between the h index and several standard bibliometric indicators and secondly we compared the h index between otolaryngologists from Europe and US. We used the Institute of Scientific Information (ISI) Web of Knowledge to identify citation reports from a random sample of influential editors from six otolaryngology journals: Journal of Laryngology and Otology (n = 21), Clinical Otolaryngology (n = 16), European Archives of Oto-Rhino-Laryngology (n = 49), The Laryngoscope (n = 66), Otolaryngology-Head and Neck Surgery (n = 15), and Archives of Otolaryngology Head and Neck Surgery (n = 15). The following data were gathered: Number of publications (P), total citations received by P(C), total citations received by P without self-citations (Cs), average number of citations per publication (CPP), and Hirsch index (h). Statistical analysis was used to correlate the above data and we also compared the h index of European and North American editors. There were 182 randomly selected editors. We observed a good correlation between the h index and other standard bibliometric indicators. Using the non-parametric Mann Whitney test, the median P between Europe and US was not statistically significant. However, the median C, CS and h were statistically significant. In conclusion, the h index is a simple yet powerful indicator as it combines productivity and impact. Overall, the US editorial panel have a higher h index. PMID- 19551397 TI - Ear, nose, and throat effects of high altitude. AB - High altitude changes human physiology and can result in illnesses such as acute mountain sickness, high-altitude cerebral edema, and high-altitude pulmonary edema. The physiological impacts of high-altitude illnesses occur secondary to extravasation of fluid from the intravascular space into the extravascular space during a rapid ascent. Headache, hearing disturbances, vestibular disturbances, epistaxis, sleep apnea, coughing, respiratory tract infections, and nasal obstruction are main ear, nose, and throat complaints of individuals travelling to high altitude. These complaints can cause delays or cancelations in a person's climbing plans. In this article, we review the ear, nose, and throat effects of high altitude based on the relevant literature. PMID- 19551399 TI - Necrotizing fasciitis of the head and neck: 34 cases of a single institution experience. AB - We analyzed 34 patients with necrotizing fasciitis (NF) of the head and neck. According to the clinical presentation, we classified NF into the cervical and cranial type. Infection can spread along the superficial lamina or along the pretracheal lamina of the deep cervical fascia. In the first type, skin necrosis is the main sequelae, while in the second case, descending infection and mediastinitis are the main complications. Of the 34 patients, in 17 the infection progressed in the mediastinum. Due to aggressive surgical therapy and postoperative intensive care, the mortality rate remained low (1/17; 6%). In this analysis, we emphasize the importance of an accurate and fast diagnosis, followed by aggressive surgical therapy and a multidisciplinary approach. PMID- 19551400 TI - Microbial DNA fingerprinting of human fingerprints: dynamic colonization of fingertip microflora challenges human host inferences for forensic purposes. AB - Human fingertip microflora is transferred to touched objects and may provide forensically relevant information on individual hosts, such as on geographic origins, if endogenous microbial skin species/strains would be retrievable from physical fingerprints and would carry geographically restricted DNA diversity. We tested the suitability of physical fingerprints for revealing human host information, with geographic inference as example, via microbial DNA fingerprinting. We showed that the transient exogenous fingertip microflora is frequently different from the resident endogenous bacteria of the same individuals. In only 54% of the experiments, the DNA analysis of the transient fingertip microflora allowed the detection of defined, but often not the major, elements of the resident microflora. Although we found microbial persistency in certain individuals, time-wise variation of transient and resident microflora within individuals was also observed when resampling fingerprints after 3 weeks. While microbial species differed considerably in their frequency spectrum between fingerprint samples from volunteers in Europe and southern Asia, there was no clear geographic distinction between Staphylococcus strains in a cluster analysis, although bacterial genotypes did not overlap between both continental regions. Our results, though limited in quantity, clearly demonstrate that the dynamic fingerprint microflora challenges human host inferences for forensic purposes including geographic ones. Overall, our results suggest that human fingerprint microflora is too dynamic to allow for forensic marker developments for retrieving human information. PMID- 19551401 TI - Tafluprost protects rat retinal ganglion cells from apoptosis in vitro and in vivo. AB - BACKGROUND: To investigate whether tafluprost, which is a prostaglandin-related compound and an anti-glaucoma drug, has a direct anti-apoptotic effect in cultured retinal ganglion cells (RGCs) and rat RGCs in retinas with optic nerve crush (ONC). METHODS: RGC-5 cells were induced to undergo apoptosis by a serum deprivation and by exogenous glutamate. The level of cell death with or without tafluprost was monitored by an XTT assay and by immunocytochemistry with activated caspase-3. Changes in intracellular calcium ([Ca(2+)]i) levels were measured with fluo-4 fluorescence. Rat RGCs were degenerated by ONC. After topical instillation of tafluprost for 7 and 14 days, the numbers of retrograde labeled RGCs were counted. Retinal flatmounts were subjected to terminal dUTP nick end labeling (TUNEL) staining to detect apoptotic cells. RESULTS: Tafluprost dose-dependently promoted RGC-5 cell viability with an optimum concentration of 3 microM (p = 0.006). Tafluprost significantly reduced caspase-3-positive cells and suppressed [Ca(+2)]i evoked by exogenous glutamate. The cGMP-dependent protein kinase inhibitor and KT-5823 partially blocked the rescue effect of tafluprost (p = 0.002). The survival rate of RGCs significantly increased in eyes treated with tafluprost (p = 0.01), and the prevalence of TUNEL-positive cells was significantly decreased 14 days after ONC (p < 0.001). CONCLUSIONS: These data suggest that tafluprost has an anti-apoptotic effect in RGCs. PMID- 19551402 TI - Acute high-intensity exercise with low energy expenditure reduced LDL-c and total cholesterol in men. AB - A reduction in LDL cholesterol and an increase in HDL cholesterol levels are clinically relevant parameters for the treatment of dyslipidaemia, and exercise is often recommended as an intervention. This study aimed to examine the effects of acute, high-intensity exercise ( approximately 90% VO(2max)) and varying carbohydrate levels (control, low and high) on the blood lipid profile. Six male subjects were distributed randomly into exercise groups, based on the carbohydrate diets (control, low and high) to which the subjects were restricted before each exercise session. The lipid profile (triglycerides, VLDL, HDL cholesterol, LDL cholesterol and total cholesterol) was determined at rest, and immediately and 1 h after exercise bouts. There were no changes in the time exhaustion (8.00 +/- 1.83; 7.82 +/- 2.66; and 9.09 +/- 3.51 min) and energy expenditure (496.0 +/- 224.8; 411.5 +/- 223.1; and 592.1 +/- 369.9 kJ) parameters with the three varying carbohydrate intake (control, low and high). Glucose and insulin levels did not show time-dependent changes under the different conditions (P > 0.05). Total cholesterol and LDL cholesterol were reduced after the exhaustion and 1 h recovery periods when compared with rest periods only in the control carbohydrate intake group (P < 0.05), although this relation failed when the diet was manipulated. These results indicate that acute, high-intensity exercise with low energy expenditure induces changes in the cholesterol profile, and that influences of carbohydrate level corresponding to these modifications fail when carbohydrate (low and high) intake is manipulated. PMID- 19551403 TI - Can the electromyographic fatigue threshold be determined from superficial elbow flexor muscles during an isometric single-joint task? AB - The purpose of this study was to compare the electromyographic fatigue threshold (EMG(FT)) values determined simultaneously from superficial elbow flexor muscles during an isometric single-joint task. Eight subjects performed isometric elbow flexions at randomly ordered percentages of maximal voluntary contraction (20, 30, 40, 50 and 60%). During these bouts, electromyographic (EMG) activity was measured in the anterior head of Deltoid, lateral head of Triceps brachii, Brachioradialis and both short and long head of Biceps brachii. For each subject and each muscle, the EMG amplitude data were plotted as function of time for the five submaximal bouts. The slope coefficient of the EMG amplitude versus time linear relationships were plotted against force level. EMG(FT) was determined as the y-intercept of this relationship and considered as valid only if the following criteria were met: (1) significant positive linear regression (P < 0.05) between force and slope coefficient, (2) an adjusted coefficient of determination for force versus slope coefficient relationship greater than 0.85, and (3) a standard error for the EMG(FT) below 5% of maximal voluntary contraction. The EMG(FT) could only be determined for one muscle (the long head of Biceps brachii) and only in three out of the eight subjects (mean value = 24.9 +/- 1.1% of maximal voluntary contraction). The lack of EMG(FT) in most of the subjects (5/8) could be explained by putative compensations between elbow muscles which were indirectly observed in some subjects. In this way, EMG(FT) should be studied from a more simple movement i.e., ideally a movement implying mainly one muscle. PMID- 19551404 TI - Time to exhaustion at maximal lactate steady state is similar for cycling and running in moderately trained subjects. AB - We compared time to exhaustion (t(lim)) at maximal lactate steady state (MLSS) between cycling and running, investigated if oxygen consumption, ventilation, blood lactate concentration, and perceived exertion differ between the exercise modes, and established whether MLSS can be determined for cycling and running using the same criteria. MLSS was determined in 15 moderately trained men (30 +/- 6 years, 77 +/- 6 kg) by several constant-load tests to exhaustion in cycling and running. Heart rate, oxygen consumption, and ventilation were recorded continuously. Blood lactate concentration and perceived exertion were measured every 5 min. t (lim) (37.7 +/- 8.9 vs. 34.4 +/- 5.4 min) and perceived exertion (7.2 +/- 1.7 vs. 7.2 +/- 1.5) were similar for cycling and running. Heart rate (165 +/- 8 vs. 175 +/- 10 min(-1); P < 0.01), oxygen consumption (3.1 +/- 0.3 vs. 3.4 +/- 0.3 l min(-1); P < 0.001) and ventilation (93 +/- 12 vs. 103 +/- 16 l min(-1); P < 0.01) were lower for cycling compared to running, respectively, whereas blood lactate concentration (5.6 +/- 1.7 vs. 4.3 +/- 1.3 mmol l(-1); P < 0.05) was higher for cycling. t(lim) at MLSS is similar for cycling and running, despite absolute differences in heart rate, ventilation, blood lactate concentration, and oxygen consumption. This may be explained by the relatively equal cardiorespiratory demand at MLSS. Additionally, the similar t(lim) for cycling and running allows the same criteria to be used for determining MLSS in both exercise modes. PMID- 19551405 TI - Pythium infection activates conserved plant defense responses in mosses. AB - The moss Physcomitrella patens (P. patens) is a useful model to study abiotic stress responses since it is highly tolerant to drought, salt and osmotic stress. However, very little is known about the defense mechanisms activated in this moss after pathogen assault. In this study, we show that P. patens activated multiple and similar responses against Pythium irregulare and Pythium debaryanum, including the reinforcement of the cell wall, induction of the defense genes CHS, LOX and PAL, and accumulation of the signaling molecules jasmonic acid (JA) and its precursor 12-oxo-phytodienoic acid (OPDA). However, theses responses were not sufficient and infection could not be prevented leading to hyphae colonization of moss tissues and plant decay. Pythium infection induced reactive oxygen species production and caused cell death of moss tissues. Taken together, these data indicate that Pythium infection activates in P. patens common responses to those previously characterized in flowering plants. Microscopic analysis also revealed intracellular relocation of chloroplasts in Pythium-infected tissues toward the infection site. In addition, OPDA, JA and its methyl ester methyl jasmonate induced the expression of PAL. Our results show for the first time JA and OPDA accumulation in a moss and suggest that this defense pathway is functional and has been maintained during the evolution of plants. PMID- 19551406 TI - DNA methylation regulates constitutive expression of Stat6 regulatory genes SOCS 1 and SHP-1 in colon cancer cells. AB - PURPOSE: Stat6 signaling is active in cancer cells and IL-4-induced Stat6 activities or Stat6 activational phenotypes vary among cancer cells. This study aimed at investigating possible mechanism(s) involved in the formation of varying Stat6 activities/phenotypes. METHODS: Stat6 regulatory genes, SOCS-1 and SHP-1, were examined for mRNA expression using RT-PCR, and their promoter DNA methylation was assayed by methylation-specific PCR in Stat6-phenotyped colon cancer cell lines. DNA methylation was then verified by sequencing. RT-PCR assay and Western blotting were used to detect the expression of SOCS-1 and SHP-1 after demethylation using 5-aza-2'-deoxycytidine. RESULTS: Compared with Stat6(null) Caco-2 cells, Stat6(high) HT-29 cells showed decreased constitutive expression of SOCS-1 and SHP-1, which correlated with DNA hypermethylation in these genes' promoters. Interestingly, demethylation in HT-29 cells recovered the constitutive expression of SOCS-1 and SHP-1. CONCLUSIONS: These findings suggest that DNA methylation controls the constitutive expression of negative Stat6 regulatory genes, which may affect Stat6 activities. PMID- 19551407 TI - Prognostic significance of a pretreatment hematologic profile in patients with head and neck cancer. AB - BACKGROUND: The purpose of this retrospective study was to test whether the pretreatment hematologic profile can predict the prognosis of patients with head and neck cancer. METHODS: Medical records from 278 patients with head and neck cancer were reviewed, and 270 cases were evaluable. Clinical data including age, gender, stage, pretreatment hematologic profile (including white blood cell, platelet, and differential counts, and hemoglobin level) were recorded. Statistical analyses were performed to determine the prognostic effect of these hematologic indicators, as well as clinical variables. The association between the hematologic indicators and clinical factors was also analyzed. RESULTS: Pretreatment monocytes >1,000 cells/microl (P = 0.028), hemoglobin <11.0 g/dl (P = 0.022), and platelet count >400 x 10(3) cells/microl (P = 0.017) were identified as independent prognostic factors in addition to the nodal status and metastasis. A significant correlation between T-stage/monocyte or platelet count and metastasis/platelet count were shown. Monocytosis, anemia, and thrombocytosis were demonstrated to have a cumulative effect on the prognosis of head and neck cancer patients (normal vs. abnormality in one lineage, P = 0.001; abnormality in one vs. more than one lineage, P = 0.005). CONCLUSIONS: A pretreatment hematologic profile can be considered as a useful prognostic marker in patients with head and neck cancer. PMID- 19551408 TI - Validity of Isospora lacazei (Labbe, 1893) infecting the house sparrow, Passer domesticus (L.), in Saudi Arabia. AB - Isospora lacazei was described from large number of passeriform birds including house sparrow (Passer domesticus). It was not certain whether all described species under this name belong to the same species. Also, two other species (Isospora passeris and Isospora passerum) were identified from P. domesticus depending on variation of oocyst measurements. During the present study, the Isospora species was reported from house sparrow (P. domesticus) in Saudi Arabia. The oocyst measurement ranges cover all the ranges of the previously described Isospora species from P. domesticus. Therefore, the present species could be identified as I. lacazei, whilst the other species being described can be considered as a junior synonym of I. lacazei. PMID- 19551409 TI - Engineering novel habitats on urban infrastructure to increase intertidal biodiversity. AB - Urbanization replaces natural shorelines with built infrastructure, seriously impacting species living on these "new" shores. Understanding the ecology of developed shorelines and reducing the consequences of urban development to fauna and flora cannot advance by simply documenting changes to diversity. It needs a robust experimental programme to develop ways in which biodiversity can be sustained in urbanized environments. There have, however, been few such experiments despite wholesale changes to shorelines in urbanized areas. Seawalls- the most extensive artificial infrastructure--are generally featureless, vertical habitats that support reduced levels of local biodiversity. Here, a mimic of an important habitat on natural rocky shores (rock-pools) was experimentally added to a seawall and its impact on diversity assessed. The mimics created shaded vertical substratum and pools that retained water during low tide. These novel habitats increased diversity of foliose algae and sessile and mobile animals, especially higher on the shore. Many species that are generally confined to lowshore levels, expanded their distribution over a greater tidal range. In fact, there were more species in the constructed pools than in natural pools of similar size on nearby shores. There was less effect on the abundances of mobile animals, which may be due to the limited time available for recruitment, or because these structures did not provide appropriate habitat. With increasing anthropogenic intrusion into natural areas and concomitant loss of species, it is essential to learn how to build urban infrastructure that can maintain or enhance biodiversity while meeting societal and engineering criteria. Success requires melding engineering skills and ecological understanding. This paper demonstrates one cost effective way of addressing this important issue for urban infrastructure affecting nearshore habitats. PMID- 19551410 TI - Imprint of past environmental regimes on structure and succession of a deep-sea hydrothermal vent community. AB - Dramatic perturbations of ecological communities through rapid shifts in environmental regime do not always result in complete mortality of residents. Instead, legacy individuals may remain and influence the succession and composition of subsequent communities. We used a reciprocal transplant experiment to investigate whether a legacy effect is detectable in communities experiencing an abrupt increase or decrease in hydrothermal fluid flux at deep-sea vents. Vent habitats are characterized by strong gradients in productivity and physico chemical stressors, both of which tend to increase with increasing vent fluid flux. In our experiments, many species survived transplantation from cool (water temperatures <2 degrees C above ambient) to warm (4-30 degrees C above ambient) habitats, resulting in significantly higher species richness on transplanted than remaining experimental substrata. A legacy effect was much less apparent in transplantation from warm to cool habitat, although a few vestimentiferan tubeworms, normally restricted to warm habitat, survived transplantation. The asymmetry in influence of legacy individuals suggests that productivity enhancement may outweigh potential physiological stress in setting limits to distributions of vent invertebrates. This influence of biological processes contrasts with theory developed in the rocky intertidal that predicts the predominance of physical control at the high-stress end of an environmental gradient. Prediction of successional transitions in vents and other habitats experiencing regime shifts in which remnant species may survive must take into account the possible influence of historical effects. PMID- 19551411 TI - Nutritional support in patients with oesophageal cancer. AB - BACKGROUND: Obesity and overweight are risk factors for developing an oesophageal cancer, especially the adenocarcinoma in the distal oesophagus or at the gastroesophageal junction, and many patients still are overweight at the clinical presentation even if they are losing weight. Main mechanisms involved in weight loss are a decreased nutrients' intake and an alteration in metabolism due to a cytokine-driven inflammatory status. Malnutrition is a risk factor for a poor compliance to chemotherapy and radiation therapy and finally for the oncologic outcome. There is scientific evidence that frequently both conditions exist but in the advanced stages of disease metabolic alterations play a major role and are responsible for the poor response to nutritional support. METHODS: The literature about the nutritional support in patients with cancer of the oesophagus has been reviewed with special emphasis on randomised clinical trials whenever available. In surgical patients, both overweight and weight loss increase the risk of postoperative complications. RESULTS: In non-dysphagic patients receiving a neo adjuvant oncologic treatment, the simple use of oral nutritional supplements is little effective in ameliorating the nutritional status, in contrast, an intensive dietetic surveillance associated with oral supplements can lead to better nutritional status, improved quality of life and better compliance with therapy. In dysphagic patients, many comparative non-randomised clinical studies have shown clinical benefits from tube feeding on the nutritional status and compliance with therapy. There is no apparent difference on the metabolic efficacy of the enteral versus parenteral nutrition. Studies on peri-operative nutrition in oesophagectomy patients were often underpowered and, hence, inconclusive, but the large experience on the nutritional support in patients with gastrointestinal cancer undergoing major abdominal surgery has clearly shown the benefits of the enteral nutrition. Both the American and the European Society for Parenteral and Enteral Nutrition have recognised a grade A recommendation for the nutritional support of malnourished gastrointestinal cancer patients undergoing major surgery. CONCLUSIONS: In patients with oesophageal cancer on chemotherapy and/or radiation therapy, enteral nutrition (oral supplements +/- intensive counselling or tube feeding) is nutritionally and clinically beneficial. In surgical patients, a broad experience in major abdominal surgery supports the peri-operative use of enteral nutrition and especially of immune enteral nutrition. PMID- 19551412 TI - New vistas in GPCR 3D structure prediction. AB - Human G-protein coupled receptors (hGPCRs) comprise the most prominent family of validated drug targets. More than 50% of approved drugs reveal their therapeutic effects by targeting this family. Accurate models would greatly facilitate the process of drug discovery and development. However, 3-D structure prediction of GPCRs remains a challenge due to limited availability of resolved structure. The X-ray structures have been solved for only four such proteins. The identity between hGPCRs and the potential templates is mostly less than 30%, well below the level at which sequence alignment can be done regularly. In this study, we analyze a large database of human G-protein coupled receptors that are members of family A in order to optimize usage of the available crystal structures for molecular modeling of hGPCRs. On the basis of our findings in this study, we propose to regard specific parts from the trans-membrane domains of the reference receptor helices as appropriate template for constructing models of other GPCRs, while other residues require other techniques for their remodeling and refinement. The proposed hypothesis in the current study has been tested by modeling human beta2-adrenergic receptor based on crystal structures of bovine rhodopsin (1F88) and human A2A adenosine receptor (3EML). The results have shown some improvement in the quality of the predicted models compared to Modeller software. PMID- 19551414 TI - Melt brushes of diblock copolymer. AB - Using self-consistent field theory (SCFT), we investigate the morphologies formed by a melt brush of AB diblock copolymers grafted to a flat substrate by their B ends. In addition to a laterally uniform morphology, SCFT predicts three ordered morphologies exhibiting different periodic patterns at the air surface: a hexagonal array of A-rich dots, an alternating sequence of A- and B-rich stripes, and a hexagonal pattern of B-rich dots. When the phase diagram of the tethered film is plotted as a function of A/B incompatibility, chiN, and diblock composition, f, it resembles the bulk phase diagram with the periodic phases converging to a mean-field critical point at weak segregation. The periodic-phase region in the phase diagram shrinks with increasing grafting density and expands when the air surface acquires an affinity for the grafted B blocks. PMID- 19551413 TI - The association of vacA genotypes and Helicobacter pylori-related gastroduodenal diseases in the Middle East. AB - The variations in the three regions of the Helicobacter pylori vacA gene, the signal (s1 and s2), intermediate (i1 and i2) and middle regions (m1 and m2), are known to cause the differences in vacuolating activities. However, it was unclear whether these vacA genotypes are associated with the development of gastric cancer and peptic ulcer in the Middle East. The aim was to identify the prevalence of vacA genotypes in the Middle East and the association with gastroduodenal diseases. We investigated the relationship of vacA genotypes to H. pylori-related disease development by meta-analysis using previous reports of 1,646 patients from the Middle East. The frequency of the vacA s1, m1 and i1 genotypes in the Middle Eastern strains was 71.5% (1,007/1,409), 32.8% (427/1,300) and 40.7% (59/145), respectively. Importantly, the frequency of vacA s- and m-region genotypes significantly differed between the north and south parts of the Middle East countries (P < 0.001). The vacA genotypes significantly increased the risk of gastric cancer (odds ratio [OR]: 4.02, 95% confidence interval [CI]: 1.98-8.14 for the s1 genotype; 2.50, 1.62-3.85 for m1; 5.27, 1.97 14.1 for s1m1; 15.03, 4.69-48.17 for i1) and peptic ulcers (OR: 3.07, 95% CI: 2.08-4.52 for s1; 1.81, 1.36-2.42 for m1). The cagA-positive genotype frequently coincided with the s1, m1 and i1 genotypes. The vacA s- and m-region genotypes may be useful risk factors for gastrointestinal diseases in the Middle East, similar to European and American countries. Further studies will be required to evaluate the effects of the i-region genotype. PMID- 19551415 TI - Fibronectin adsorption studied using neutron reflectometry and complementary techniques. AB - In implantology it is known that fibronectin affects cell-substrate adhesion, consequently, the structure and composition of the initially adsorbed fibronectin layer to a large extent determines the biological response to a biomaterial implanted into the body. In this study we have used neutron reflectometry and quartz-crystal microbalance with dissipation to investigate the amount of fibronectin adsorbed, the layer density, thickness and structure of films adsorbed to polished silicon oxide surfaces. We have cultured MG63 osteoblast like cells on surfaces coated and uncoated with fibronectin and monitored the cellular response to these surfaces. The results show that at fibronectin concentrations in the range 0.01 to 0.1 mg/ml a single highly hydrated layer of fibronectin approximately 40-50 A in thickness adsorbs to a polished silicon oxide surface and is likely to correspond to one diffuse monolayer of fibronectin arranged side-on. Cells cultured on this fibronectin layer have dramatically different morphology and growth to those grown on bare surfaces. Using a model silicon oxide surface has enabled us to study the substrate/protein interface, together with the impact of a fibronectin layer on the cellular response using consistent experimental conditions across a unique set of experimental techniques. PMID- 19551416 TI - Spreading of diblock copolymer droplets: a probe of polymer micro-rheology. AB - We present an experimental study of the spreading dynamics of symmetric diblock copolymer droplets above and below the order-disorder transition. Disordered diblock droplets are found to spread as a homopolymer and follow Tanner's law (the radius grows as R approximately t(m), where t is time and m = 1/10). However, droplets that are in the ordered phase are found to be frustrated by the imposed lamellar microstructure. This frustration is likely at the root of the observed deviation from Tanner's law: droplet spreading has a much slower power law (m approximately 0.05+/-0.01). We show that the different spreading dynamics can be reconciled with conventional theory if a strain-rate-dependent viscosity is taken into account. PMID- 19551417 TI - The impact of environmental changes upon the microrheological response of adherent cells. AB - The mechanical behaviour of adherent cells cultured in vitro is known to be dependent on the mechanical properties of the substrate. We show that this mechanical behaviour is also strongly affected by the cells' environment. We focus here on the impact of temperature and pH. Experiments carried out on individual cells in a tuneable environment reveal that the intra-cellular mechanical behaviour exhibits large and fast changes when the external cell environment is changed. Fast passive microrheometry measurements allow for the precise characterisation of the transient regime observed during a temperature drop. When maintained at a non-physiological temperature, the cells reach a stabilised state distinct from the state observed in physiological conditions. The perturbation can be reversed but exhibits hysteretic behaviour when physiological conditions are restored. The transient regime observed during the recovery process is found to be different from the transient regime observed when leaving physiological conditions. A modified generalized Stokes-Einstein equation taking into account the cell activity through an effective temperature is proposed here to fit the experimental results. Excellent agreement between the model and the measurements is obtained for time lags from 10-3 to 1 s considered in this study. PMID- 19551418 TI - Kinetic arrest in polyion-induced inhomogeneously charged colloidal particle aggregation. AB - Polymer chains adsorbed onto oppositely charged colloidal particles can significantly modify the particle-particle interactions. For sufficient amounts of added polymers, the original electrostatic repulsion can even turn into an effective attraction and relatively large aggregates can form. The attractive interaction contribution between two particles arises from the correlated adsorption of polyions at the oppositely charged particle surfaces, resulting in a non-homogeneous surface charge distribution. Here, we investigate the aggregation kinetics of polyion-induced colloidal complexes through Monte Carlo simulation, in which the effect of charge anisotropy is taken into account by a DLVO-like inter-particle potential, as recently proposed by Velegol and Thwar (Langmuir 17, 7687 (2001)). The results reveal that the aggregation process slows down due to the progressive increase of the potential barrier height upon clustering. Within this framework, the experimentally observed cluster phases in polyelectrolyte-liposome solutions can be interpreted as a kinetic arrested state. PMID- 19551419 TI - Localization and delocalization of energy in a Peyrard-Bishop chain. AB - This paper studies energy localization conditions in lattices of the type proposed by Peyrard and Bishop. Homogeneous and inhomogeneous lattices are analyzed and the role of interfaces in the latter is emphasized. Simulations allowed us to identify critical energy values for the existence of localization. After a certain energy value, it is possible to observe the loss of energy localization along the chain. PMID- 19551420 TI - Critical dynamics of lateral and transversal phase separations in bilayer biomembranes and surfactants. AB - We consider bilayer biomembranes or surfactants made of two chemically incompatible amphiphile molecules, which may laterally or transversely phase separate into macrodomains, upon variation of some suitable parameter (temperature, lateral pressure, etc.). The purpose is an extensive study of the dynamics of both lateral and transverse phase separations, when the bilayer is suddenly cooled down from a high initial temperature towards a final one very close to the spinodal point. The critical dynamics are investigated through the partial dynamic structure factors of different species. Using a two-order parameter field theory, where the two fields are the composition fluctuations of one component in the leaflets of the bilayer, combined with an extended van Hove approach that is based on two coupled Langevin equations (with noise), we exactly compute these dynamic structure factors. We first find that the dynamics is governed by two time scales. The longest one, Tau, can be related to the thermal correlation length, Xi ~ Sigma|T - T(c)|(-1/2), by Tau ~ Xi(z), with the dynamic critical exponent z = 4, where Sigma is an atomic length scale, T the absolute temperature, and T(c) its critical value. The characteristic time Tau can be interpreted as the time required for the formation of the final macrophase domains. The second time scale is rather shorter, and can be viewed as the short time during which the unlike phospholipids execute local motion. Second, we demonstrate that the dynamic structure factors obey exact scaling laws, and depend on three lengths, namely the wavelength q(-1) (q is the wave vector modulus), the correlation length Xi, and a length scale R(t) ~ t(1/z) (z = 4) representing the size of macrophase domains at time t. Of course, the two lengths Xi and R(t) coincide at the final time Tau at which the bilayer reaches its final equilibrium state. Finally, the present work must be considered as a natural extension of our previously published one dealing with the study of lateral and transverse phase separations from a static point of view. PMID- 19551421 TI - [Myalgia of the masticatory muscles]. AB - Masticatory muscle pain can be regarded as a regional manifestation of musculoskeletal disorders similar to those observed in other body regions. Along with temporomandibular joint pain and some painless disturbances related to mandibular mobility they are subsumed under the term temporomandibular disorders (TMD). Masticatory muscle pain is assumed to be associated with a variety of biophysiological risk factors. Valid diagnostic instruments make it possible to differentiate between the various TMD subgroups. In most cases, masticatory muscle pain can be treated/managed successfully. In a considerable number of patients, however, the pain persists over a long period of time despite therapeutic interventions. Understanding of the underlying neurobiological background of acute and chronic pain may help in therapeutic decision-making and evaluation of the therapeutic effects. PMID- 19551425 TI - Small bowel obstruction due to a right-sided paraduodenal hernia: a case report. AB - Small bowel obstruction (SBO) diagnosed with abdominal computed tomography (CT) has been extensively studied in radiology literature. We present a case report of SBO due to a rare right-sided paraduodenal hernia diagnosed preoperatively on a non-contrast CT and confirmed surgically. PMID- 19551426 TI - Gallbladder torsion diagnosed by MDCT and MRCP. PMID- 19551427 TI - The gastric band: first-choice procedure for obesity surgery. AB - The advent of laparoscopic adjustable gastric banding (LAGB) during the latter part of the 20th century represents a watershed in the management of chronic obesity. In this paper we provide an overview of LAGB with respect to its development, clinical outcomes, and future role. We also address current controversies, including a comparison of LAGB with Roux-en-Y gastric bypass (RYGBP). At present LAGB seems to be increasing in popularity in the United States, whereas in Europe there seems to be a trend away from gastric banding toward RYGBP. Optimal outcomes after LAGB are a function of correct laparoscopic technique, an experienced surgical team, a well-engineered device, and intensive long-term follow-up. The majority of studies show that LAGB is an extremely safe and effective procedure, with an operative mortality of 0-0.1% and excess weight loss (%EWL) of 50-60%. Commensurate with this degree of weight loss, almost all studies show substantial improvements in obesity-related co-morbidities, such as hypertension, type II diabetes, and dyslipidemia. In addition, LAGB has been shown to be both safe and effective in the super-obese, in adolescents, and in older patients and can be delivered as an ambulatory procedure. Operative mortality and early complication rates are significantly higher for RYGBP and, whilst gastric bypass results in greater weight loss than LAGB in the first 2 years, at 3 years and beyond the difference appears to be less marked. Overall, LAGB provides a safe, effective intervention for obese patients and remains our first-choice procedure for bariatric surgery. PMID- 19551428 TI - Candida isolation in patients hospitalized on a surgical ward: significance and mortality-related factors. AB - BACKGROUND: Infections caused by Candida are an emerging pathology on surgical wards. The aim of the present study was to evaluate the incidence, characteristics, and predictive factors of mortality in patients colonized and/or infected by Candida spp. in this setting. METHODS: A consecutive series of 105 patients hospitalized on a general surgery ward between 2000 and 2004 were included, and 118 positive cultures for Candida were identified. The variables age, sex, previous medical history, current disease, anemia, ICU stay, type and localization of the microorganism, need for parenteral nutrition, and transfusions were recorded. The primary outcome was in-hospital mortality. A univariate analysis was performed to determine which of these variables were associated with mortality. With a logistic regression model, independent prognostic factors of mortality were determined. RESULTS: The prevalence of patients colonized and/or infected by Candida on our surgical ward was 0.98% (CI 95%: 0.79-1.17), and the incidence was 49 cases per 1,000 patient-years. Of the 105 patients in this series, 56 were men (53%) and 49 women (47%); the mean age was 63.8 years (SD +/- 15.7). Twelve patients (11.4%) had candidemia. Crude mortality was 23% (24 patients), whereas the mortality attributable to candidemia was 25% (3/12 cases). Anemia (p = 0.001); transfusions (p = 0.003), and an ICU stay (p = 0.002) were associated with mortality. Candidemia was associated with neoplasms (p = 0.02) and the infection caused by Candida parapsilosis (p = 0.04). The only independent factor related to mortality was the anemia (p = 0.028; Odds Ratio: 6.43; 95% CI: 1.23-33.73). CONCLUSIONS: Colonization and/or infection by Candida spp. in non-ICU hospitalized surgical patients implies a relative high mortality. Anemia is an independent factor for mortality. PMID- 19551429 TI - Increase in the serum bile acid level predicts the effective hypertrophy of the nonembolized hepatic lobe after right portal vein embolization. AB - BACKGROUND: The purpose of the present study was to investigate the clinical association between serum bile acid level changes and liver hypertrophy in portal vein embolization (PVE). METHODS: In 31 patients, the serum total bile acid level was prospectively measured before and 1, 3, 5, 7, and 14 days after right PVE. Computed tomographic volumetry was performed before and 25.0 +/- 3.6 days after PVE. RESULTS: Portal vein embolization induced the liver hypertrophy with a median increase in the left lobe volume (ILV) of 165 cm(3) and a median percentage ILV (%ILV) of 29%. Compared with the pretreatment level, the serum bile acid levels significantly increased on day 3 and day 14 after PVE (p = 0.017 and p = 0.003, respectively). In patients with greater hypertrophy after PVE (ILV > 165 cm(3) and %ILV > 30%), the increases in the bile acid level on day 3 were larger than that in those with lesser hypertrophy (p = 0.008 and p = 0.002, respectively). The increase on day 3 positively correlated with the ILV and %ILV (p = 0.003 and p = 0.004, respectively). The serum bile acid levels on day 3, 5, and 7 after PVE increased in patients with %ILV > 30% but not in those with %ILV < or = 30%. CONCLUSIONS: Portal vein embolization increases the serum bile acid level in patients with effective liver hypertrophy in the nonembolized lobe. The increase on day 3 is a useful predictor of effective hypertrophy of the nonembolized lobe. Thus, bile acid signaling may be important for liver regeneration post-PVE. PMID- 19551430 TI - Anatomical changes after inguinal hernia treatment: a reason for chronic pain and recurrent hernia? AB - BACKGROUND: Chronic pain and hernia recurrence are the most frequent long-term complications of treating inguinal hernia. One reason may be postsurgical changes in the anatomy of the groin. METHODS: In a retrospective investigation from 1994 to 2008, 1,194 patients undergoing 1,421 laparoscopic transabdominal preperitoneal (TAPP) herniorrhaphies were studied. Anatomical structures in the groin, seen in 1,214 primary and 207 recurrent hernias, were charted by means of video analysis. Hernia orifices, myopectineal orifice (MPO), and Hesselbach's and Hessert's triangles were measured in their respective vertical and horizontal diameters in order to calculate the surface area. Other anatomical changes were also recorded. RESULTS: The mean surface area of hernial orifices was 3.00 +/- 2.01 cm(2) in primary hernias and 3.60 +/- 3.81 cm(2) in recurrent hernias. The mean surface area of Hesselbach's triangle was 4.23 +/- 2.21 cm(2) in the former group and 2.09 +/- 2.10 cm(2) in the latter (p < 0.0001). The mean surface area of Hessert's triangle in primary hernias (9.03 +/- 6.17 cm(2)) was significantly larger than that in recurrent hernias (3.11 +/- 3.67 cm(2); p < 0.0001). Further anatomical changes in suture-treated recurrent hernias included a dislocated spermatic cord, a raised inguinal ligament, and asymmetry in the region. CONCLUSION: The treatment of inguinal hernia by the suture technique is followed by significant anatomical changes such as reduction of the surface area and a subsequent increase of tension in the inguinal region. This could be one of the main reasons for chronic pain and hernia recurrence. PMID- 19551431 TI - Is a bougie required for the performance of the fundal wrap during laparoscopic Nissen fundoplication? AB - BACKGROUND: It has been claimed that oesophageal intubation with a bougie during laparoscopic Nissen fundoplication (LNF) reduces the risk of a tight crural repair and wrap, and thereby decreases the prevalence of post-operative dysphagia. The aim of this study is to assess the benefit of routinely inserting a bougie during LNF, in relation to post-operative dysphagia. METHODS: All patients who underwent LNF by a single surgeon between March 2005 and March 2007 were reviewed. Oesophageal intubation during surgery was routinely performed in all patients who underwent LNF during the first 11 months of the study period, whilst during the second phase, routine oesophageal intubation was not performed. The main outcome measures were the prevalence of post-operative dysphagia and complication rates. Dysphagia severity was assessed clinically by a scoring system at discharge, and again at 6 weeks, 3 months, 6 months and 1 year. RESULTS: Forty patients had a bougie inserted (group 1) and 42 had no bougie (group 2). The mean age was 49.1 (SD, standard deviation +/- 8.1) years in group 1 and 49.4 (SD +/- 8.4) years in group 2 (p = 0.88). There were no major complications. When assessed at 12 weeks, 60% of group 1 and 51% of group 2 patients still had some degree of dysphagia (p = 0.635) but by 1 year dysphagia had resolved in all patients (p = 1.00). There was no significant difference in the median dysphagia scores between the two groups at: discharge (p = 0.181), 6 weeks (p = 0.234), 12 weeks (p = 0.504), 24 weeks (p = 0.182) or 1 year (p = 0.530). Analysis of the progression of dysphagia over the first post-operative year using Cox regression analysis did not show any significant difference between the two groups (p = 0.375). CONCLUSIONS: LNF can be safely performed without the routine use of an oesophageal bougie and this practice does not result in increased post-operative dysphagia rates. PMID- 19551432 TI - Effectiveness of stent placement for palliative treatment in malignant colorectal obstruction and predictive factors for stent occlusion. AB - BACKGROUND: Self-expandable metallic stent (SEMS) for malignant colorectal obstruction is widely used in palliative treatment and as an alternative to surgery. The aims of this study are to evaluate the effectiveness of stent placement for palliative treatment and to identify the predictive factors associated with stent occlusion. METHODS: A retrospective analysis was performed in 55 patients who had undergone placement of an uncovered SEMS from February 2004 to April 2007 for palliative treatment of malignant colorectal obstruction with metastatic or locally advanced cancer that was surgically unresectable. We analyzed the technical and clinical outcomes of stent placement, complications related to the procedure, stent patency rate, and predictive factors associated with stent occlusion. RESULTS: The causes of colorectal obstruction before stent placement were primary colorectal cancer in 42 patients and noncolorectal extrinsic cancer in 13 patients. The initial technical success rate was 98.2%, and the clinical success rate was 94.4%. Complications occurred in 17 patients (30.9%). These included stent occlusion (n = 8), migration (n = 6), bowel perforation (n = 1), stent distortion (n = 1), and fistula formation (n = 1). The mean and median stent patency periods were 184 days [95% confidence interval (CI), 137-230 days] and 141 days (95% CI, 69-213 days), respectively. The degree of expansion 48 h after stent placement was significantly better in the nonocclusion group than in the stent occlusion group. In the multivariate Cox proportional hazard model, insufficient stent expansion (<70%) 48 h after stent placement was significantly associated with an increase in stent occlusion during the follow-up period (odds ratio, 12.55; p = 0.002). CONCLUSIONS: Uncovered SEMS placement is an effective palliative treatment for malignant colorectal obstruction. The degree of stent expansion 48 h after stent placement is significantly associated with the maintenance of stent patency and is a predictive factor for stent occlusion. PMID- 19551433 TI - Laparoscopic cholecystectomy in cirrhotic patients: the value of MELD score and Child-Pugh classification in predicting outcome. AB - BACKGROUND: Laparoscopic cholecystectomy is a challenging procedure in patients with cirrhosis. This study aims to evaluate the safety and outcome of laparoscopic cholecystectomy in patients with cirrhosis and examines the value of model for end-stage liver disease (MELD) score and Child-Pugh classification in predicting morbidity. MATERIALS AND METHODS: From January 1995 to July 2008, 220 laparoscopic cholecystectomies were performed in cirrhotic, Child-Pugh class A and B patients. Indications included symptomatic gallbladder disease and cholecystitis. MELD score ranged between 8 and 27. Child-Pugh class and MELD score were preoperatively calculated and associated with postoperative results. Data regarding patients and surgical outcome were retrospectively analyzed. RESULTS: No deaths occurred. Postoperative morbidity occurred in 19% of the patients and included hemorrhage, wound complications, and intra-abdominal collections controlled conservatively. Intraoperative difficulty due to liver bed bleeding was experienced in 19 patients. Conversion to open cholecystectomy was necessary in 12 cases. Median operative time was 95 min. Median hospital stay was 4 days. Patients with preoperative MELD score above 13 showed a tendency for higher complication rate postoperatively. Child-Pugh classification did not seem to predict morbidity effectively. CONCLUSION: Laparoscopic cholecystectomy can be performed safely in selected patients with cirrhosis Child-Pugh A and B and symptomatic cholelithiasis with acceptable morbidity. Some of its advantages are shorter operative time and reduced hospital stay. MELD score seems to predict morbidity more accurately than Child-Pugh classification system. PMID- 19551434 TI - Establishing construct validity of a virtual-reality training simulator for hysteroscopy via a multimetric scoring system. AB - BACKGROUND: The aims of this study are to determine construct validity for the HystSim virtual-reality (VR) training simulator for hysteroscopy via a new multimetric scoring system (MMSS) and to explore learning curves for both novices and experienced surgeons. METHODS: Fifteen relevant metrics had been identified for diagnostic hysteroscopy by means of hierarchical task decomposition. They were grouped into four modules (visualization, ergonomics, safety, and fluid handling) and individually weighted, building the MMSS for this study. In a first step, 24 novice medical students and 12 experienced gynecologists went through a self-paced teaching tutorial, in which all participants received clearly stated goals and instructions on how to carry out hysteroscopic procedures properly for this study. All subjects performed five repeated trials on two different exercises on HystSim (exploration and diagnosis exercises). After each trial the results were presented to the participants in the form of an automated objective feedback report (AOFR). Construct validity for the MMSS and learning curves were investigated by comparing the performance between novices and experienced surgeons and in between the repeated trials. To study the effect of repeated practice, 23 of the novices returned 2 weeks later for a second training session. RESULTS: Comparing novices with the experienced group, the ergonomics and fluid handling modules resulted in construct validity, while the visualization module did not, and for the safety module the experienced group even scored significantly lower than novices in both exercises. The overall score showed only construct validity when the safety module was excluded. Concerning learning curves, all subjects improved significantly during the training on HystSim, with clear indication that the second training session was beneficial for novice surgeons. CONCLUSIONS: Construct validity for HystSim has been established for different modules of VR metrics on a new MMSS developed for diagnostic hysteroscopy. Careful refinement and further testing of metrics and scores is required before using them as assessment tools for operative skills. PMID- 19551435 TI - Preventing and managing complications of laparoscopic gastrectomy. PMID- 19551436 TI - A systematic review of methods to palliate malignant gastric outlet obstruction. AB - BACKGROUND: The traditional approach to palliating patients with malignant gastric outlet obstruction (GOO) has been open gastrojejunostomy (OGJ). More recently endoscopic stenting (ES) and laparoscopic gastrojejunostomy (LGJ) have been introduced as alternatives, and some studies have suggested improved outcomes with ES. The aim of this review is to compare ES with OGJ and LGJ in terms of clinical outcome. METHOD: A systematic literature search and review was performed for the period January 1990 to May 2008. Original comparative studies were included where ES was compared with either LGJ or OGJ or both, for the palliation of malignant GOO. RESULTS: Thirteen studies met the inclusion criteria (10 retrospective cohort studies, two randomised controlled trials and one prospective study). Compared with OGJ, ES resulted in an increased likelihood of tolerating an oral intake [odds ratio (OR) 2.6, p = 0.02], a shorter time to tolerating an oral intake (mean difference 6.9 days, p < 0.001) and a shorter post-procedural hospital stay (mean difference 11.8 days, p < 0.001). There were no significant differences between 30-day mortality, complication rates or survival. There were an inadequate number of cases to quantitatively compare ES with LGJ. CONCLUSION: This review demonstrates improved clinical outcomes with ES over OGJ for patients with malignant GOO. However, there is insufficient data to adequately compare ES with LGJ, which is the current standard for operative management. As these conclusions are based on observational studies only, future large well-designed randomised controlled trials (RCTs) would be required to ensure the estimates of the relative efficacy of these interventions are valid. PMID- 19551437 TI - Minimally invasive colon resection for malignant colonic conditions is associated with a transient early increase in plasma sVEGFR1 and a decrease in sVEGFR2 levels after surgery. AB - INTRODUCTION: Plasma VEGF levels increase after minimally invasive colorectal resection (MICR) and remain elevated for 2-4 weeks. VEGF induces physiologic and pathologic angiogenesis by binding to endothelial cell (EC) bound VEGF-Receptor-1 (VEGFR1) and VEGFR2. Soluble forms of these receptors sequester plasma VEGF, decreasing the amount available to bind to EC-bound receptors. Ramifications of surgery-related plasma VEGF changes partially depend on plasma levels of sVEGFR1 and sVEGFR2. This study assessed perioperative sVEGFR1 and sVEGFR2 levels after MICR in patients with colorectal cancer. METHODS: Forty-five patients were studied; blood samples were taken from all patients preoperatively (preop) and on postoperative days (POD) 1 and 3; in most a fourth sample was drawn between POD 7 30. Late samples were bundled into two time points: POD 7-13 and POD 14-30. sVEGFR1 and sVEGFR2 levels were measured via ELISA. sVEGFR2 data are reported as mean +/- SD and were assessed with the paired samples t test. sVEGFR1 data were not normally distributed. They are reported as median and 95% confidence interval (CI) and were assessed with the Wilcoxon signed-Rank test (p < 0.05). RESULTS: Preoperatively, the mean plasma sVEGFR2 level (7583.9 pg/ml) was greater than the sVEGFR1 result (98.3 pg/ml). Compared with preop levels, sVEGFR2 levels were significantly lower on POD 1 (6068.2 pg/ml, +/-2034.5) and POD 3 (6227.6 pg/ml, +/-2007.0), whereas sVEGFR1 levels were significantly greater on POD 1 (237.5 pg/ml; 95% CI, 89.6-103.5), POD 3 (200.2 pg/ml; 95% CI, 159-253), and POD 7-13 (102.9 pg/ml; 95% CI, 189.7-253). No differences were found on POD 7-13 for sVEGFR2 or POD 14-30 for either protein. CONCLUSIONS: sVEGFR2 values decreased and sVEGFR1 levels increased early after MICR; sVEGFR2 changes dominate due to their much larger magnitude. The net result is less plasma VEGF bound by soluble receptors and more plasma VEGF available to bind to ECs early after surgery. PMID- 19551438 TI - Laparoscopy: a safe approach to appendicitis during pregnancy. AB - BACKGROUND: The aim of this study was to evaluate laparoscopic versus open surgery for suspected appendicitis during pregnancy. METHODS: A hospital-based retrospective review of 65 consecutive pregnant patients who underwent surgery for suspected appendicitis from 1999 to 2008 was performed. Significance was determined by Pearson's chi(2) test, Fisher's exact test, Mann-Whitney test, and Kruskal-Wallis test. RESULTS: Of the 65 patients, 48 cases were laparoscopic and 17 open. Use of the laparoscopic versus open approach was significantly increased in the first (100% vs. 0%, p < 0.001) and second trimesters (73% vs. 27%, p < 0.001). The open approach was used more frequently in third-trimester patients (71% vs. 29%, p = NS). Significance was demonstrated in mean length of hospital stay in the laparoscopic versus open group (3.4 vs. 4.2 days, p = 0.001). No maternal mortalities occurred. Follow-up of fetal outcome was achieved in 89% of patients. No difference was demonstrated in fetal loss (1 in laparoscopic group), APGAR score, birth weight, and preterm delivery rate by operative approach. Adverse outcome was associated with maternal temperature greater than 38 degrees C, leukocytosis greater than 16 x 10(9)/l, or more than 48 h between onset of symptoms and emergency room presentation. CONCLUSIONS: This article is the largest hospital-based series evaluating the laparoscopic versus open approach for pregnant patients with presumed acute appendicitis. While methodological limitations preclude a definitive recommendation, laparoscopy appears to be a safe, feasible, and efficacious approach for pregnant patients with presumed acute appendicitis. We conclude that it is likely not the surgical approach but the underlying diagnosis combined with maternal factors that determine the risk for pregnancy complications. A benefit of laparoscopy is the diagnostic ability to identify other intra-abdominal pathology which may mimic appendicitis and harbor pregnancy risks. PMID- 19551439 TI - Current technique of laparoscopic total mesorectal excision (TME): an international questionnaire among 368 surgeons. AB - BACKGROUND: Current literature shows no consensus for the technique of laparoscopic total mesorectal excision (LTME). This study aimed to assess the current practice of LTME. METHODS: From January to March 2008, members of the European Association for Endoscopic Surgery (EAES), the Indian Association of Gastrointestinal Endo-Surgeons (IAGES), and the Society of Laparoscopic Surgeons (SLS), together with renowned surgeons in the field of LTME, were invited to fill out an online questionnaire concerning aspects of LTME. RESULTS: The 368 questionnaires showed that 77% of the study participants performed 1-20 LTMEs per year (low volume) and that 33% performed more than 20 LTMEs per year (high volume). Preoperative bowel preparation (PBP), Trendelenburg position, periumbilical insertion of a 30o laparoscope, medial-to-lateral dissection, ultrasonic hemostasis, high-tie ligation, splenic flexure mobilization, left ureteral identification, partial sigmoid resection, extraction of the specimen by a new minilaparotomy and wound protector, end-to-end stapled anastomosis using a 28- to 29-mm anvil with 3.5-mm staples, abdominal lavage, pelvic drainage, and diverting ileostoma were performed by a majority of the surgeons. Less consistency was observed in identification of the right ureter, dissection of Denonvilliers' fascia, location of the minilaparotomy, and construction of a colonic pouch. There were significant differences between high and low volume and between American and European surgeons. Significantly more low-volume surgeons indicated a preference for an open TME depending on the age and gender of the patient, the presence of comorbidity, previous laparotomy, and locally advanced tumor. More low-volume surgeons applied PBP (83.4% vs. 71.8%; p = 0.017). On the average, high-volume surgeons identified more autonomic pelvic nerves during dissection (2.6 vs. 1.8 nerves). The right ureter was identified by 66% of the American and 31.2% of the European surgeons. In the United States 91.5% and in Europe 61.2% created an end-to-end anastomosis. Pouches were created by 32% of the European and 6.8% of the American surgeons. CONCLUSION: The respondents showed an apparent preference for several aspects of LTME. Differences were related to expertise and still more to continent. PMID- 19551440 TI - The impact of proton-pump inhibitors on intraperitoneal sepsis: a word of caution for transgastric NOTES procedures. AB - BACKGROUND: During transgastric natural orifice transluminal endoscopic surgery (NOTES), there is an iatrogenic perforation of the gastric wall with leakage of gastric contents into the peritoneal cavity. The aim of this study is to determine the effect of proton-pump inhibitors (PPI) and alterations of gastric pH on infection during transgastric surgery. METHODS: Thirty 250-g male Sprague Dawley rats were divided into a study group (SG, n = 15) and a control group (CG, n =15). SG were given 5 mg/kg pantoprazole for 3 days before procedure and another dose 1 h before. CG received saline at similar time points. A mini laparotomy with gastrotomy was performed. Aspiration of 2.0 cc gastric contents was removed from the stomach and injected into the peritoneal cavity of both groups. Gastric pH and peritoneal pH levels were obtained. Gastric aspirate was sent for culture. White blood cell counts (WBC) were obtained on postoperative days 1, 7, and 14, and C-reactive protein (CRP) levels were obtained on postoperative day 1. At day 14, a necropsy was performed and aerobic and anaerobic cultures of the peritoneal cavity were obtained. RESULTS: There were no deaths in either group. The average gastric pH in the SG was 5.13 versus 3.26 (p = 0.03) in the CG. The average peritoneal pH was similar in both groups. The WBC in the SG was 4.5 vs. 3.5 (1,000 cells/mm) in the CG. There was no elevation in CRP levels in either group. Bacterial cultures were positive in 3/15 (20%) rats in the CG and in 9/15 (60%) in the SG (p = 0.008). Intra-abdominal abscesses were found in 2/15 (13%) rats in the CG and in 5/15 (33%) in the SG (p = 0.08). CONCLUSIONS: Pretreatment with a PPI resulted in a higher rate of peritoneal bacterial contamination and abscess formation. The acidic environment of the stomach appears to be protective against infection when intraperitoneal contamination occurs as a result of gastrotomy. PMID- 19551441 TI - Prospective randomized comparison of clinical results between hand-assisted laparoscopic and open splenectomies. AB - BACKGROUND: Although some studies have compared laparoscopic and hand-assisted laparoscopic splenectomy (HALS) in splenomegaly cases, no study has analyzed the differences between HALS and open splenectomy (OS). This study aimed to compare the HALS and OS techniques in splenomegaly cases. METHODS: This prospective study included 27 patients undergoing splenectomy for splenic disorders at the Department of General Surgery, Istanbul Medical Faculty between February and October 2007. Open splenectomy was performed for 14 patients and HALS for the remaining 13 patients. RESULTS: The end points compared included incision length, operative time, intraoperative blood loss, postoperative drain output and duration, postoperative pain scores, length of postoperative hospitalization, and perioperative complications. The authors found benefits of HALS over OS for incision length, postoperative pain score, postoperative drain output and duration, and hospital stay. The main advantages of the HALS technique over OS were less postoperative pain (p = 0.0002), shorter hospital stay (p = 0.004), and shorter abdominal incision (p = 0.012). CONCLUSIONS: For splenomegaly, HALS significantly facilitates the surgical procedure and reduces the hospital stay while maintaining the advantages of OS such as tactile sense as well as easy and atraumatic manipulation of enlarged spleens. PMID- 19551442 TI - Laparoscopic Heller myotomy for achalasia: results after 10 years. AB - BACKGROUND: Laparoscopic Heller myotomy was first undertaken in the early 1990s, and appreciable numbers of patients with 10-year follow-up periods are now available. This study was undertaken to determine long-term outcomes after laparoscopic Heller myotomy used to treat achalasia. METHODS: Of 337 patients who have undergone laparoscopic Heller myotomy since 1992, 47 who underwent myotomy more than 10 years ago have been followed through a prospectively maintained registry. Among many symptoms, patients scored dysphagia, chest pain, vomiting, regurgitation, choking, and heartburn before and after myotomy using a Likert scale with choices ranging from 0 (never/not bothersome) to 10 (always/very bothersome). Symptom scores before and after myotomy were compared using a Wilcoxon matched-pairs test. Data are reported as median (mean +/- standard deviation). RESULTS: The median length of the hospital stay was 2 days (mean, 3 +/- 8.6 days; range, 1-60 days). Notable complications were infrequent after myotomy. There were no perioperative deaths. One patient required a redo myotomy after 5 years due to recurrence of symptoms. At this writing, 33 patients (70%) are still alive. The causes of death after discharge were unrelated to myotomy. The frequency and severity scores for dysphagia, chest pain, vomiting, regurgitation, choking, and heartburn all decreased significantly after laparoscopic Heller myotomy (p < 0.0001 for all). CONCLUSIONS: Laparoscopic Heller myotomy can be undertaken with few complications. This procedure significantly decreases the frequency and severity of achalasia symptoms without promoting heartburn. The symptoms of achalasia are durably ameliorated by laparoscopic Heller myotomy during long-term follow-up evaluation, thereby promoting application of this procedure. PMID- 19551443 TI - Charcoal suspension tattoo localization for differentiated thyroid cancer recurrence. AB - BACKGROUND: The high sensitivity of ultrasound and thyroglobulin determination for follow-up of differentiated thyroid cancer allows early detection of nonpalpable recurrences. Intraoperative localization of these small foci in previously dissected necks is a surgical challenge. We describe a new technique for ultrasound-guided tattooing to facilitate excision. METHODS: Prospective study of 15 consecutive patients with suspected recurrence of differentiated carcinoma. Whole-body scan after administration of 100 mCi (131)I, performed in 14 cases, was negative in 13. TSH stimulated thyroglobulin averaged 31 ng/ml (<1 182 ng/ml). During ultrasound 19 lesions were discovered in regions already addressed by en bloc neck dissection. Lymph node metastasis was confirmed by cytology in 11 and by washout thyroglobulin in 2. Fine-needle aspiration (FNA) was insufficient for analysis in 1 and was not performed for 5 because of the size (<5 mm). Colloidal charcoal (1-4 ml) was injected under ultrasound, 1-15 days preoperatively. Tolerance, intraoperative charcoal localization, and success of resection were recorded. RESULTS: The injection was well tolerated. Charcoal was found in or just next to 16 lesions (84%). In 1 case it was found several centimeters away. In 1 case, no charcoal was found. In 1 case, hematoma caused by injection impaired surgical exploration. Surgery removed 18 lesions (95%) in 14 patients (93%): carcinoma (16), benign lymphadenitis (2). CONCLUSIONS: Ultrasound guided charcoal tattooing is safe, easy, and well-tolerated for localization of nonpalpable lesions in previously operated necks, with a high rate of success. Excision of these small recurrences remains controversial, however, and may not impact survival or quality of life. PMID- 19551444 TI - Outcome of locally recurrent and metastatic angiosarcoma. AB - BACKGROUND: Angiosarcoma (AS) is a rare soft tissue sarcoma with an enhanced propensity for local and systemic failure. The outcome of locally recurrent and metastatic AS treated at a single institution was evaluated. METHODS: Medical records of AS patients treated for local recurrence and distant metastasis (1993 2008) were retrospectively reviewed. Univariable and multivariable analyses were performed to identify prognosticators. RESULTS: Forty-four patients were treated for locally recurrent AS; the majority (59%) were 5 cm as the only independent adverse prognosticator of recurrent AS-specific survival [hazard ratio (HR): 3.26, P = 0.04]. Ninety-nine patients were treated for metastatic AS; 73% had multiple metastatic sites; the lung was the most common site (36%). Chemotherapy, mainly doxorubicin- and/or paclitaxel-based regimens, were administered to 95 patients (96%). Radiotherapy was utilized in 25% cases; 16% of patients underwent curative intent surgery. Median DSS was 10 months (95% CI: 7.9-12 months). Isolated lymph node metastasis versus hematogenic spread was the only statistically significant favorable prognostic factor identified (HR: 0.29, P = 0.01). CONCLUSION: Locally recurrent AS is often treatable; complete resection can potentially prolong survival. In contrast, metastatic patients have a grave prognosis; however, patients with isolated lymphatic spread and possibly those treated with taxol based chemotherapeutic regimens have a favorable outcome. PMID- 19551445 TI - Rapamycin inhibits hepatectomy-induced stimulation of metastatic tumor growth by reduction of angiogenesis, microvascular blood perfusion, and tumor cell proliferation. AB - BACKGROUND: Liver regeneration after hepatectomy stimulates metastatic tumor growth through the release of potent growth factors. In the signaling cascades of several growth factors, mTOR is a downstream mediator, triggering cell survival and cell cycle progression. The mTOR inhibitor rapamycin (RAPA) has been shown to exhibit potent antitumor activities. However, it is unknown whether RAPA is capable of exerting these effects under the conditions of hepatectomy-associated liver regeneration. We therefore analyzed the effect of RAPA and cyclosporine A (CyA) on tumor growth characteristics after major hepatectomy using a mouse model of colorectal metastasis. METHODS: Tumor growth was studied by using GFP transfected CT26.WT colorectal cancer cells, which were implanted into the dorsal skinfold chambers of BALB/c-mice after 70% hepatectomy. The animals were treated daily with RAPA (1.5 mg/kg) or CyA (10 mg/kg). Tumors were analyzed for angiogenesis, microvascular blood perfusion, cell proliferation, apoptotic cell death, and tumor growth. RESULTS: RAPA significantly inhibited tumor growth compared with CyA and sham treatment. This was associated with a decreased microvascular density within the tumors and a markedly reduced microvascular blood perfusion. CyA only slightly reduced angiogenesis and tumor growth. The effects of RAPA were associated with a significant reduction of tumor cell proliferation, whereas manifestation of apoptotic cell death was not affected by the immunosuppressive treatment regimen. CONCLUSIONS: RAPA is capable of inhibiting angiogenesis, microvascular blood perfusion, and tumor growth of colorectal metastasis during hepatectomy-associated liver regeneration. Thus, targeting mTOR might represent an interesting strategy to prevent tumor recurrence after hepatectomy for colorectal metastasis. PMID- 19551446 TI - Comparison of gene-transfer efficiency in human embryonic stem cells. AB - Technologies designed to allow manipulation and modification of human embryonic stem (hES) cells are numerous and vary in the complexity of their methods, efficiency, reliability, and safety. The most commonly studied and practiced of these methods include electroporation, lipofection, nucleofection, and lentiviral transduction. However, at present, it is unclear which protocol offers the most efficient and reliable method of gene transfer to hES cells. In this study, a bi fusion construct with ubiquitin promoter driving enhanced green fluorescent protein reporter and the firefly luciferase (pUb-eGFP-Fluc) along with neomycin selection marker was used for in vitro and in vivo studies. In vitro studies examined the transfection efficiency and viability of each technique using two hES cell lines (male H1 and female H9 cells). Lentiviral transduction demonstrated the highest efficiency (H1: 25.3 +/- 4.8%; H9: 22.4 +/- 6.5%) with >95% cell viability. Nucleofection demonstrated transfection efficiency of 16.1 +/- 3.6% (H1) and 5.8 +/- 3.2% (H9). However, minimal transfection efficiency was observed with electroporation (2.1 +/- 0.4% (H1) and 1.9 +/- 0.3% (H9)) and lipofection (1.5 +/- 0.5% (H1) and 1.3 +/- 0.2% (H9); P < 0.05 vs. lentiviral transduction). Electroporation also demonstrated the highest cell death (62 +/- 11% (H1) and 42 +/- 10% (H9)) followed by nucleofection (25 +/- 9% (H1) and 30 +/ 15 (H9)). Importantly, lentiviral transduction generated a greater number of hES cell lines stably expressing the double-fusion reporter gene (hES-DF) compared to other transfection techniques. Finally, following subcutaneous transplantation into immunodeficient nude mice, the hES-eGFP-Fluc cells showed robust proliferation as determined by longitudinal bioluminescence imaging. In summary, this study demonstrates that lentiviral transduction and nucleofection are efficient, simple, and safe techniques for reliable gene transfer in hES cells. The double-fusion construct provides an attractive approach for generating stable hES cell lines and monitoring engraftment and proliferation in vitro and in vivo. PMID- 19551447 TI - Comparison of medicine resident diabetes care between Veterans Affairs and academic health care systems. AB - BACKGROUND: Although others have reported national disparities in the quality of diabetes care between the Veterans Affairs (VA) and private health care delivery systems, it is not known whether these differences persist among internal medicine resident providers in academic settings. OBJECTIVE: We compared the quality of diabetes primary care delivered by resident physicians in either a private academic health care system (AHS) or its affiliated VA health care system. DESIGN: Cross-sectional observational study PARTICIPANTS: We included patients who: had a diagnosis of diabetes, had >2 primary care visits with the same resident provider during 2005, and were not separately managed by an attending physician or endocrinologist. A total of 640 patients met our criteria and were included in the analysis. MEASUREMENTS AND RESULTS: Compared to the VA, patients in the AHS were more likely to be younger, female, have fewer medications, and be treated with insulin, but had less comorbidity. Patients in the VA were more likely to be referred for an annual eye exam (94% vs. 78%), receive lipid screening (88% vs. 74%), receive proteinuria screening (63% vs. 34%), and receive a complete foot exam (85% vs. 32%) in analyses adjusted for patient demographics and comorbidities (p-value <0.001 for all comparisons). In adjusted analyses, there were no significant differences in HbA1(c), blood pressure, or LDL cholesterol control. CONCLUSIONS: In spite of similar resident providers and practice models, there were substantial differences in the diabetes quality of care delivered in the VA and AHS. Understanding how these factors influence subsequent practice patterns is an important area for study. PMID- 19551448 TI - Internal medicine resident self-report of factors associated with career decisions. AB - BACKGROUND: Little is known about factors contributing to the career decisions of internal medicine residents. OBJECTIVE: To evaluate factors self-reported by internal medicine residents nationally as important to their career decisions. DESIGN: Cross-sectional survey conducted in October of 2005, 2006, and 2007 as part of the national Internal Medicine In-Training Examination (IM-ITE). PARTICIPANTS: Postgraduate year 3 internal medicine residents taking the IM-ITE. MEASUREMENTS: Residents rated the importance of nine factors in their career decisions on 5-point Likert scales. Univariate statistics characterized the distribution of responses. Associations between variables were evaluated using Cochran-Mantel-Haenszel statistics for ordinal data. Multivariate analyses were conducted using logistic regression. RESULTS: Of 17,044 eligible residents taking the IM-ITE, 14,890 (87.4%) completed the career decision survey questions. Overall, time with family was the factor most commonly reported as of high or very high importance to career decisions (69.6%). Women were more likely to assign greatest importance to family time (OR 1.22, 95% confidence interval 1.12 1.31, p < 0.001) and long-term patient relationships (OR 1.34, 95% confidence interval 1.23-1.46, p < 0.001). Across debt levels, financial considerations were of greatest importance more often for residents owing >$150,000 (OR 1.33, 95% confidence interval 1.09-1.62, p < 0.001). Across specialties, mentor specialty was rated lowest in importance by residents pursuing hospitalist and general internal medicine careers. CONCLUSIONS: Greater attention to factors reported by residents as important to their career decisions may assist efforts to optimize the distribution of physicians across disciplines. In addition to lifestyle and practice considerations, these factors may include mentor specialty. As this factor is less commonly reported as important by residents planning careers in generalist fields, attention to effective mentoring may be an important element of efforts to increase interest in these areas. PMID- 19551449 TI - Population-based fracture risk assessment and osteoporosis treatment disparities by race and gender. AB - BACKGROUND: Undertreatment of osteoporosis has been recognized as a common problem in selected patient subgroups. However, primary prevention has been hampered by limited risk assessment tools that can be applied to large populations. OBJECTIVES: Using clinical risk factors with a new tool from the World Health Organization (FRAX) and recommendations from the National Osteoporosis Foundation (NOF), we evaluated fracture risk and osteoporosis treatment in a US cohort. PARTICIPANTS: African Americans and Caucasians recruited from 2003-7 across the US as part of a longitudinal study. DESIGN: Cross-sectional. MEASURES: The number of persons receiving prescription osteoporosis medications was assessed by race, sex, and fracture risk. Multivariable logistic regression evaluated the association between receipt of osteoporosis medications and fracture risk after controlling for potential confounders. RESULTS: Among 24,783 participants, estimated fracture risk was highest for Caucasian women. After multivariable adjustment for fracture-related risk factors, the likelihood of receipt of osteoporosis medications among African Americans was lower than among Caucasians [odds ratio (OR) = 0.44, 95% confidence interval (CI) 0.37, 0.53] and for men compared to women (OR = 0.08, 95% CI 0.06 0.10). Even for the highest risk group, Caucasian women with 10-year hip fracture risk > or = 3% (n = 3,025, 39.7%), only 26% were receiving treatment. CONCLUSIONS: A substantial gap exists between 2008 NOF treatment guidelines based on fracture risk and the receipt of prescription osteoporosis medications. This gap was particularly notable for African Americans and men. FRAX is likely to be useful to assess risk at a population level and identify high-risk persons in need of additional evaluation. PMID- 19551450 TI - Inhibition of p38 MAPK facilitates ex vivo expansion of skin epithelial progenitor cells. AB - Ex vivo expansion of skin epithelial stem cells has long attracted great interest because of the potential utilization in transplantation and gene therapy. The use of cultured stem or progenitor cells was limited by the lack of applicable culturing system with both satisfactory expansion efficacy and well suppressed differentiation ex vivo. The p38 mitogen-activated protein kinase (MAPK) pathways are responsible for cell growth and differentiation process. We investigated the function of p38 inhibitor SB203580 in the ex vivo expansion of skin epithelial progenitor cells by comparing media with or without addition of this inhibitor. Our results showed that the culturing medium with murine 3T3 feeder layers added with 10 microM SB203580 was more effective in promoting clonal growth of human skin epithelial progenitors or stem cells than the conventional medium without SB203580. The clone initial day in cells treated with 10 microM SB203580 came 2 d earlier with higher colony formation efficiency. The skin epithelial progenitor cells treated with 10 microM SB203580 formed clones that were uniformly smaller in size, longer in sustained proliferation, shorter in clone doubling time, higher in S-phase cells percentage, and lower in levels of differentiation markers such as K10 along with higher levels of stem-cell-associated markers such as p63, K15, and ABCG2 than those cultured in the conventional medium. Collectively, these results indicate that the p38 MAPK pathways inhibitor SB203580 can be used as a culture medium additive that helps to achieve more effective ex vivo expansion of skin epithelial progenitor cells. PMID- 19551452 TI - Single incision transumbilical laparoscopic Roux-en-Y gastric bypass: a first case report. AB - Roux-en-Y gastric bypass, which is based on the dual mechanisms of restriction and mal-absorption, is considered to be one of the gold standard surgeries for treatment of morbid obesity. However, the classic laparoscopic approach requires five to seven incisions for multiple trocar placement. Recently, single incision laparoscopic surgery has been adopted for performing appendectomies, cholecystectomies, sleeve gastrectomies, and adjustable gastric band surgeries. Here, we described the first case of a patient receiving laparoscopic Roux-en-Y gastric bypass through a single transumbilical incision. The operative time was 170 min. There were no intra-operative complications; the patient did very well postoperatively and was discharged 2 days later. Single incision laparoscopic surgery has been viewed as an alternative to natural orifice transluminal endoscopic surgery. When performed via the transumbilical route, it can make the abdominal wound scarless and cosmetically more acceptable. PMID- 19551451 TI - Transforming growth factor-beta1 (TGF-beta1) and acetylcholine (ACh) alter nitric oxide (NO) and interleukin-1beta (IL-1beta) secretion in human colon adenocarcinoma cells. AB - Colon adenocarcinoma is one of the most common fatal malignancies in Western countries. Progression of this cancer is dependent on tumor microenvironmental signaling molecules such as transforming growth factor-beta (TGF-beta) or acetylcholine (ACh). The present study was conducted to assess the influence of recombinant human transforming growth factor (rhTGF)-beta1 or ACh on nitric oxide (NO) and interleukin-1beta (IL-1beta) secretion by three human colon adenocarcinoma cell lines: HT29, LS180, and SW948, derived from different grade tumors (Duke's stage). The cells were cultured in 2D and 3D (spheroids) conditions. Colon carcinoma cells exhibited different sensitivities to rhTGF beta1 or ACh dependent on the tumor grade and the culture model. ACh exhibited significant inhibitory effects towards NO, endothelial nitric oxide synthase (eNOS), and IL-1beta secretion especially by tumor cells derived form Duke's C stage of colon carcinoma. rhTGF-beta1 also decreased NO, IL-1beta, and eNOS expression, but its effect was lower than that observed after the administration of ACh. The inhibition of NO and IL-1beta production was more striking in 3D tumor spheroids than in 2D culture monolayers. Taken together, the TGF-beta1-ACh axis may regulate colon carcinoma progression and metastasis by altering NO secretion and influence inflammatory responses by modulating IL-1beta production. PMID- 19551453 TI - Successful medical management of status post-Roux-en-Y-gastric-bypass hyperinsulinemic hypoglycemia. AB - Roux-en-Y gastric bypass (RYGBP) is the most commonly performed type of bariatric surgery, which is used in the treatment of obesity and type 2 diabetes. Recent case reports and case series have described a rare complication of RYGBP, status post-gastric-bypass hyperinsulinemic hypoglycemia, which was mainly managed successfully with pancreatectomy. In this letter, we describe the first successful management of status post-gastric-bypass hyperinsulinemic hypoglycemia with diazoxide. PMID- 19551454 TI - An evaluation of the rapid airway management positioner in obese patients undergoing gastric bypass or laparoscopic gastric banding surgery. AB - A new positioning device, the Rapid Airway Management Positioner (RAMP, Airpal Inc., Center Valley, PA) was evaluated to determine if there was an improvement in either mask ventilation, direct laryngoscopy, or both with the use of the RAMP in this patient population. Fifty-one morbidly obese patients (BMI > 35 kg/m(2)) undergoing elective bariatric surgery were enrolled. Ventilation and laryngoscopy was performed in the neutral and head-elevated laryngoscopy position (HELP). Direct laryngoscopy was performed noting the glottic view according to the Cormack-Lehane classification (Samsoon and Young, Anesthesiology 42:487, 1987). Mask ventilation was then recommenced. The HELP, or "ramped," position was achieved by inflating the RAMP, which was placed underneath the patient prior to entering the OR. Once proper HELP position was achieved, a second laryngoscopy was performed followed by endotracheal intubation. Two main outcomes were noted in the neutral and HELP positions: (1) laryngoscopic view and (2) ease of ventilation. The inflated ramped position provided greater ease of ventilation as compared to the neutral position (p = 0.0003). There was also a significant improvement in the glottic view in the ramped position (p = 0.04). Ease of intubation was perceived to be severely difficult among two, and overall use of the positioning device was found to be difficult among seven of the residents. The RAMP effectively positions morbidly obese patients in the HELP position. Ease of ventilation and laryngoscopic view were both improved with its use in this patient population. PMID- 19551455 TI - CXCR4 and CXCL12 expression is increased in the nigro-striatal system of Parkinson's disease. AB - Except for a handful of inherited cases related to known gene defects, Parkinson's disease (PD) is a sporadic neurodegenerative disease of unknown etiology. There is increasing evidence that inflammation and proliferation of microglia may contribute to the neuronal damage seen in the nigro-striatal dopaminergic system of PD patients. Microglia events that participate in neuronal injury include the release of pro-inflammatory and neurotoxic factors. Characterizing these factors may help to prevent the exacerbation of PD symptoms or to remediate the disease progression. In rodents, the nigro-striatal system exhibits high expression of the chemokine receptor CXCR4. Its natural ligand CXCL12 can promote neuronal apoptosis. Therefore, the present study investigated the expression of CXCR4 and CXCL12 in post-mortem brains of PD and control (non PD) individuals and in an animal model of PD. In the human substantia nigra (SN), CXCR4 immunoreactivity was high in dopaminergic neurons. Interestingly, the SN of PD subjects exhibited higher expression of CXCR4 expression and CXCL12 than control subjects despite the loss of dopamine (DA) neurons. This effect was accompanied by an increase in activated microglia. However, results from post mortem brains may not provide indication as to whether CXCL12/CXCR4 can cause the degeneration of DA neurons. To examine the role of these chemokines, we determined the levels of CXCL12 and CXCR4 in the SN of MPTP-treated mice. MPTP produced a time-dependent up-regulation of CXCR4 that preceded the loss of DA neurons. These results suggest that CXCL12/CXCR4 may participate in the etiology of PD and indicate a new possible target molecule for PD. PMID- 19551458 TI - A modified protocol for bisulfite genomic sequencing of difficult samples. AB - The bisulfite genomic sequencing protocol is a widely used method for analyzing DNA methylation. It relies on the deamination of unmethylated cytosine residues to uracil; however, its high rates of DNA degradation and incomplete cytosine to uracil conversion often lead to failed experiments, uninformative results, and false positives. Here, we report the addition of a single-step multiple restriction enzyme digestion (MRED) designed to differentially digest polymerase chain reaction products amplified from unconverted DNA while leaving those of converted DNA intact. We show that for our model system, RARB2 P2 promoter, use of MRED increased informative sequencings ninefold, and MRED did not alter the clonal representation in one fully methylated cell line, H-596, treated or not with 5-azadeoxycytidine, a methylation inhibitor. We believe that this method may easily be adapted for analyzing other genes and provide guidelines for selecting the most appropriate MRED restriction enzymes. PMID- 19551457 TI - Withania somnifera prevents morphine withdrawal-induced decrease in spine density in nucleus accumbens shell of rats: a confocal laser scanning microscopy study. AB - Opiate withdrawal is associated with morphological changes of dopamine neurons in the ventral tegmental area and with reduction of spine density of second-order dendrites of medium size spiny neurons in the nucleus accumbens shell but not core. Withania somnifera has long been used in the Middle East, Africa, and India as a remedy for different conditions and diseases and a growing body of evidence points to its beneficial effects on a number of experimental models of neurological disorders. Recently, many studies focused on the potential neuritic regeneration and synaptic reconstruction properties of its methanolic extract and its constituents (withanolides). This study investigates whether morphine withdrawal-induced spine reduction in the nucleus accumbens is affected by the administration of a Withania somnifera extract. To this end, rats were chronically treated with Withania somnifera extract along with morphine or saline and, upon spontaneous (1 and 3 days) or pharmacologically precipitated withdrawal, their brains were fixed in Golgi-Cox stain for confocal microscopic examination. In a separate group of animals, Withania somnifera extract was administered during three days of spontaneous withdrawal. Withania somnifera extract treatment reduced the severity of the withdrawal syndrome when given during chronic morphine but not during withdrawal. In addition, treatment with Withania somnifera extract during chronic morphine, but not during withdrawal, fully prevented the reduction of spine density in the nucleus accumbens shell in spontaneous and pharmacologically precipitated morphine withdrawal. These results indicate that pretreatment with Withania somnifera extract protects from the structural changes induced by morphine withdrawal potentially providing beneficial effects on the consequences related to this condition. PMID- 19551459 TI - Use of the lactose-[13C]ureide breath test for diagnosis of small bowel bacterial overgrowth: comparison to the glucose hydrogen breath test. AB - PURPOSE: The glucose hydrogen breath test (GHBT) is commonly used as a noninvasive test to diagnose small bowel bacterial overgrowth (SBBO) but its validity has been questioned. Our aim was to evaluate the lactose-[(13)C]ureide breath test (LUBT) to diagnose SBBO and to compare it with the GHBT, using cultures of intestinal aspirates as a gold standard. METHODS: In 22 patients with suspected SBBO (14 male, age range 18-73 years) aspirates were taken from the region of the ligament of Treitz under sterile conditions and cultured for bacterial growth. More than 10(6) colony-forming units/mL fluid or the presence of colonic flora was defined as culture positive (c+). After oral intake of 50 g glucose and 2 g of lactose-[(13)C]ureide, end-expiratory breath samples were obtained up to 120 min. The (13)C/(12)C ratio in breath CO(2) was determined by isotope ratio-mass spectrometry and hydrogen concentration in breath was analyzed electrochemically. RESULTS: After analyzing receiver operating characteristic curves of the LUBT results, total label recovery of >0.88% at 120 min was considered positive. The test had a sensitivity of 66.7% and a specificity of 100% to predict c+. In the GHBT, an increase of the signal of > or =12 ppm from baseline was considered positive. The sensitivity and specificity of the test were 41.7 and 44.4%, respectively. CONCLUSIONS: The new stable isotope-labeled LUBT has excellent specificity but suboptimal sensitivity. In contrast, the standard GHBT lacks both high sensitivity and specificity. The LUBT is superior to the GHBT for detecting SBBO. PMID- 19551456 TI - Role of synucleins in Alzheimer's disease. AB - Alzheimer's disease (AD) and Parkinson's disease (PD) are the most common causes of dementia and movement disorders in the elderly. While progressive accumulation of oligomeric amyloid-beta protein (Abeta) has been identified as one of the central toxic events in AD leading to synaptic dysfunction, accumulation of alpha synuclein (alpha-syn) resulting in the formation of oligomers has been linked to PD. Most of the studies in AD have been focused on investigating the role of Abeta and Tau; however, recent studies suggest that alpha-syn might also play a role in the pathogenesis of AD. For example, fragments of alpha-syn can associate with amyloid plaques and Abeta promotes the aggregation of alpha-syn in vivo and worsens the deficits in alpha-syn tg mice. Moreover, alpha-syn has also been shown to accumulate in limbic regions in AD, Down's syndrome, and familial AD cases. Abeta and alpha-syn might directly interact under pathological conditions leading to the formation of toxic oligomers and nanopores that increase intracellular calcium. The interactions between Abeta and alpha-syn might also result in oxidative stress, lysosomal leakage, and mitochondrial dysfunction. Thus, better understanding the steps involved in the process of Abeta and alpha syn aggregation is important in order to develop intervention strategies that might prevent or reverse the accumulation of toxic proteins in AD. PMID- 19551460 TI - Efficient production of ectoine using ectoine-excreting strain. AB - Halophilic bacteria strain Halomonas salina DSM 5928 was found to excrete ectoine, suggesting its potential in the development of a new method of ectoine production. We performed HPLC and LC-MS analyses that showed that Halomonas salina DSM 5928 excreted ectoine under constant extracellular osmolarity. Medium adopting monosodium glutamate as a sole source of carbon and nitrogen was beneficial for ectoine synthesis. The total concentration of ectoine was not affected by NaCl concentration in the range 0.5-2 mol l(-1). The total concentration of ectoine and productivity in a 10-l fermentor with 0.5 mol l(-1) NaCl were 6.9 g l(-1) and 7.9 g l(-1) d(-1), respectively. These findings show that Halomonas salina DSM 5928 efficiently produces ectoine at relatively low NaCl concentration. This research also indicates the potential application of free or immobilized cells for continuous culture to produce ectoine. PMID- 19551461 TI - Eye of the tiger-like MRI in parkinsonian variant of multiple system atrophy. AB - Parkinsonian variant of multiple system atrophy (MSA-P) clinically presents as autonomic dysfunction with parkinsonian features. Parkinsonian features include bradykinesia, rigidity, tremor, postural instability and poor levo-dopa response. Neuropathologically, MSA-P is characterized by selective neuronal loss and gliosis mainly affecting the putamen and caudate nucleus, substantia nigra, olivopontocerebellar pathway and intermediolateral cell column of the spinal cord. Therefore, the target of magnetic resonance imaging (MRI) is focused on signal changes or volume reduction on putamen, including putaminal slit, gliosis by diffusion studies and reduction of putaminal volume. There have been no reports describing clinical manifestations of MSA-P with imaging abnormalities over globus pallidus. Here, we describe three patients with typical presentations of MSA-P with autonomic dysfunction and disturbances of axial motor function with minimal appendicular symptoms, including postural instability and gait difficulties. MRI showed symmetrical hyperintensity over the center of globus pallidus surrounded by a mild low-signal rims at T2-weighted image that is similar to that of eye of the tiger sign except for the marked hypointense rims. Dopamine transporter scans showed symmetric reduction of uptake over bilateral basal ganglia. This is the first report concerning these unusual imaging findings in MSA-P patients and we believe there is a subgroup of MSA-P with clinical presentation of axial impairment and symmetrically abnormal signal changes of globus pallidus in MRI. PMID- 19551463 TI - Penis: a 'sanctuary' site of extramedullary leukemia relapse. PMID- 19551462 TI - Olfactory identification in non-demented elderly population and in mild cognitive impairment: a comparison of performance in clinical odor identification versus Boston Naming Test. AB - Performance in olfactory identification was studied in mild cognitive impairment (MCI), using slightly expanded standard clinical approach to study the olfactory nerve. Four hundred and eighty-six cognitively normal individuals and 72 individuals with MCI underwent spontaneous and cued odor identification and delayed odor recall. Performance in these was compared with the performance in the CERAD version of the Boston Naming Test (BNT). The individuals with MCI scores significantly worse in all tests compared with controls, but the performance in tests assessing odor were less impaired than performance in the BNT. Standard assessment of olfactory nerve function is not sufficient to study cognitive impairment in MCI. PMID- 19551464 TI - Multiple myeloma presenting with acquired factor VIII inhibitor. AB - An initial presentation of hematological malignancies associated with autoantibodies is not common, and there is only one documented case of multiple myeloma presenting with acquired FVIII inhibitor for multiple myeloma. In this paper, we describe a second case of multiple myeloma who presented with acquired FVIII inhibitor. A 43-year-old woman was referred to our hematology unit for anemia and an elevated erythrocyte sedimentation rate. Two months before her admission, she had undergone an operation at a local hospital because of ovarian cyst rupture complicated by severe postoperative bleeding. Because coagulation tests had revealed a prolonged partial thromboplastin time which could not be corrected by a mixing test and a decreased FVIII level, a diagnosis of acquired FVIII inhibitor had been made. The patient was hospitalized in our unit for further evaluation. The erythrocyte sedimentation rate was 110 mm/h, serum albumin level 2.5 g/dL, globulin level 5.6 g/dL, and C-reactive protein 47.8 mg/L (0-6). Serum IgG was high, and serum protein electrophoresis showed a monoclonal spike in the gamma region. An IgG-kappa paraprotein was identified by immunofixation of the urine and serum. X-ray films of the bones revealed lytic areas in the skull, pelvis, and lumbar vertebrae. Bone marrow aspiration showed normal cellularity with 40% plasma cell infiltration. The patient was diagnosed with the IgG kappa type of multiple myeloma associated with acquired FVIII inhibitor. In patients presenting with severe bleeding, autoantibodies against FVIII should be considered for the differential diagnosis of bleeding. Clinicians should be alert to the presence of rare underlying neoplastic diseases such as multiple myeloma, in patients with acquired FVIII inhibitor. PMID- 19551465 TI - Phase II study of neoadjuvant treatment with doxorubicin, docetaxel, and capecitabine (ATX) in locally advanced or inflammatory breast cancer. AB - BACKGROUND: Pathologic complete response (pCR) after preoperative systemic chemotherapy (PSCh) is associated with better outcome in locally advanced breast cancer (LABC). PATIENTS AND METHODS: PSCh included: doxorubicin (A) 50 mg/m(2) i.v. on day 1; docetaxel (T) 30 mg/m(2) i.v. on days 1, 8 and 15; and capecitabine (X) 1,500 mg/m(2)/day p.o. on days 1-14, in a 4-week course repeated for up to four cycles (ATX), followed by surgery. The primary end point of this study was to evaluate the pCR rate. Secondary endpoints included clinical response rate, disease-free survival (DFS), overall survival (OS), and the toxicity profile. RESULTS: A total of 60 patients were included in the analysis. Median age was 49 years, and 63.3% of patients were hormone receptor positive. The median number of cycles of PSCh was four (95% CI: 3-4). Five patients (8.3%) achieved pCR in both breast and nodes, and 16.7% reached pCR only in nodes. The clinical response rate was 77% (27% complete response), but only 18% of the patients underwent conservative surgery. With a median follow-up of 20 months, 3 year DFS and OS were 76 and 90%, respectively. Grade III/IV toxicity included neutropenia (74%), febrile neutropenia (9%), mucositis (12%), and diarrhea (12%). CONCLUSIONS: ATX every 28 days for four cycles is associated with a modest activity (low pCR rate) in the neoadjuvant setting of LABC. PMID- 19551466 TI - Role of the metabotropic glutamate receptor subtype 1 in the harmaline-induced tremor in rats. AB - The present study shows that JNJ 16259685-a selective antagonist of glutamate metabotropic receptor subtype 1 (mGluR1) injected in doses of 0.16 and 0.32 mg/kg sc strongly enhances tremor of forelimbs, head and trunk, hypolocomotion, and ataxia induced by harmaline (7.5 mg/kg ip) in rats. JNJ 16259685 inhibited locomotor and exploratory activity per se. These results may suggest an inhibitory influence of mGluR1 on the harmaline-induced motor disturbances. PMID- 19551468 TI - NAP protects against cyanide-related microtubule destruction. AB - The peptide NAP (NAPVSIPQ) was shown to protect neurons against a wide variety of insults. Particularly, NAP was shown to be neuroprotective in vitro against cyanide in hippocampal cultures and against oxygen-glucose deprivation in hippocampal and cortical neuronal cultures. Cyanide causes energy depletion in the cell and destroys the cytoskeleton, and NAP has been shown before to protect the microtubule cytoskeleton. The current study explored the effect of NAP on cyanide-induced microtubule destruction in cerebral cortical cultures. Sodium cyanide (6.8 mM) reduced the number of neurons containing intact microtubules as identified by bIII-tubulin immunostaining. When sodium cyanide was added together with NAP (10(-14)-10(-12) M), complete protection was observed. Although the primary site of action of cyanide is considered to be the mitochondria, the current results involve microtubule destruction by cyanide toxicity that is completely reversed by NAP treatment. PMID- 19551469 TI - Clustering and spatial correlations of the neuronal cytoplasmic inclusions, astrocytic plaques and ballooned neurons in corticobasal degeneration. AB - This study tested three hypotheses: (1) that there is clustering of the neuronal cytoplasmic inclusions (NCI), astrocytic plaques (AP) and ballooned neurons (BN) in corticobasal degeneration (CBD), (2) that the clusters of NCI and BN are not spatially correlated, and (3) that the lesions are correlated with disease 'stage'. In 50% of the regions, clusters of lesions were 400-800 mum in diameter and regularly distributed parallel to the tissue boundary. Clusters of NCI and BN were larger in laminae II/III and V/VI, respectively. In a third of regions, the clusters of BN and NCI were negatively spatially correlated. Cluster size of the BN in the parahippocampal gyrus (PHG) was positively correlated with disease 'stage'. The data suggest the following: (1) degeneration of the cortico-cortical pathways in CBD, (2) clusters of NCI and BN may affect different anatomical pathways and (3) BN may develop after the NCI in the PHG. PMID- 19551467 TI - Neuroprotective effects of probenecid in a transgenic animal model of Huntington's disease. AB - Huntington's disease (HD) is an autosomal dominantly inherited disorder, caused by an expanded polyglutamine region of a protein called huntingtin. The excitotoxicity, oxidative damage and altered membrane transport may have an important role in the pathogenesis of HD. Probenecid is a non-selective inhibitor of multidrug resistance-associated proteins, but it also inhibits organic anion transporters. In this study, we examined the effects of probenecid on the survival, behaviour and immunohistochemical changes in the N171-82Q transgenic mouse model of HD. After probenecid administration, the duration of survival improved by 35%. The motor activity was significantly ameliorated as compared with the control transgenic group. Probenecid treatment significantly reduced the neuronal loss and the number of neuronal intranuclear aggregates. These results suggest that probenecid may exert a neuroprotective effect by increasing the membrane transport of protective compounds, and/or inhibiting the toxic compounds. PMID- 19551470 TI - The complete nucleotide sequence of the barley yellow dwarf GPV isolate from China shows that it is a new member of the genus Polerovirus. AB - The complete nucleotide sequence of the ssRNA genome of a Chinese GPV isolate of barley yellow dwarf virus (BYDV) was determined. It comprised 5673 nucleotides, and the deduced genome organization resembled that of members of the genus Polerovirus. It was most closely related to cereal yellow dwarf virus-RPV (77% nt identity over the entire genome; coat protein amino acid identity 79%). The GPV isolate also differs in vector specificity from other BYDV strains. Biological properties, phylogenetic analyses and detailed sequence comparisons suggest that GPV should be considered a member of a new species within the genus, and the name Wheat yellow dwarf virus-GPV is proposed. PMID- 19551472 TI - Clinical trials in heart failure: may we trust? PMID- 19551471 TI - Alleviating perinatal depressive symptoms and stress: a nurse-community health worker randomized trial. AB - To determine whether a Nurse-Community Health Worker (CHW) home visiting team, in the context of a Medicaid enhanced prenatal/postnatal services (EPS), would demonstrate greater reduction of depressive symptoms and stress and improvement of psychosocial resources (mastery, self-esteem, social support) when compared with usual Community Care (CC) that includes Medicaid EPS delivered by professionals. Greatest program benefits were expected for women who reported low psychosocial resources, high stress, or both at the time of enrollment. Medicaid eligible pregnant women (N = 613) were randomly assigned to either usual CC or the Nurse-CHW team. Mixed effects regression was used to analyze up to five prenatal and postnatal psychosocial assessments. Compared to usual CC, assignment to the Nurse-CHW team resulted in significantly fewer depressive symptoms, and as hypothesized, reductions in depressive symptoms were most pronounced for women with low psychosocial resources, high stress, or both high stress and low resources. Outcomes for mastery and stress approached statistical significance, with the women in the Nurse-CHW group reporting less stress and greater mastery. Women in the Nurse-CHW group with low psychosocial resources reported significantly less perceived stress than women in usual CC. No differences between the groups were found for self-esteem and social support. A Nurse-CHW team approach to EPS demonstrated advantage for alleviating depressive symptoms in Medicaid eligible women compared to CC, especially for women at higher risk. PMID- 19551473 TI - Ascites due to pericardial cyst. PMID- 19551474 TI - Future drugs for migraine. AB - Migraine is a complex, neurovascular disorder in which genetic and environmental factors interact. At present, frontline therapies in the acute treatment of migraine include the use of non-steroidal anti-inflammatory drugs and triptans. Evidence indicates that calcitonin gene-related peptide (CGRP) plays a fundamental role in the mechanism of migraine. CGRP is a strong vasodilatatory neuropeptide that is released from activated trigeminal sensory nerves. The development of CGRP antagonists has also been driven by the fact that triptans are vasoconstrictive and cannot be safely used in patients with cardiovascular risk factors. Olcegepant (BIBN4096) is the first CGRP antagonist for the treatment of migraine that has been tested in clinical trials, but because of its poor oral bioavailability, only the intravenous formulation has been tested. The first oral non-peptide CGRP antagonist, telcagepant, has been shown recently to be highly effective in the treatment of migraine attacks. This development can be considered as the most important pharmacological breakthrough for migraine treatment since the introduction of sumatriptan in the early 1990s. These results are also of importance, since they support an interesting pathophysiological hypothesis of migraine. The pipeline of future compounds for the treatment of acute migraine headaches include TPRV1 antagonists, prostaglandin E receptor 4 (EP(4)) receptor antagonists, serotonin 5HT1(F) receptor agonists and nitric oxide synthase inhibitors. The immediate future of a preventative treatment for migraine headaches is well represented by botulinum toxin type-A, glutamate NMDA receptor antagonists, gap-junction blocker tonabersat and an angiotensin type 1 blocker candesartan. PMID- 19551475 TI - Cloning and characterization of two novel chloroplastic glycerol-3-phosphate dehydrogenases from Dunaliella viridis. AB - Dunaliella, a unicellular green alga, has the unusual ability to survive dramatic osmotic stress by accumulating high concentrations of intracellular glycerol as a compatible solute. The chloroplastic glycerol-3-phosphate dehydrogenase (GPDH) has been considered to be the key enzyme that produces glycerol for osmoregulation in Dunaliella. In this study, we cloned the two most prominent GPDH cDNAs (DvGPDH1 and DvGPDH2) from Dunaliella viridis, which encode two polypeptides of 695 and 701 amino acids, respectively. Unlike higher plant GPDHs, both proteins contained extra phosphoserine phosphatase (SerB) domains at their N termini in addition to C-terminal GPDH domains. Such bi-domain GPDHs represent a novel type of GPDH and are found exclusively in the chlorophyte lineage. Transient expression of EGFP fusion proteins in tobacco leaf cells demonstrated that both DvGPDH1 and DvGPDH2 are localized in the chloroplast. Overexpression of DvGPDH1 or DvGPDH2 could complement a yeast GPDH mutant (gpd1Delta), but not a yeast SerB mutant (ser2Delta). In vitro assays with purified DvGPDH1 and DvGPDH2 also showed apparent GPDH activity for both, but no SerB activity was detected. Surprisingly, unlike chloroplastic GPDHs from plants, DvGPDH1 and DvGPDH2 could utilize both NADH and NADPH as coenzymes and exhibited significantly higher GPDH activities when NADH was used as the coenzyme. Q-PCR analysis revealed that both genes exhibited transient transcriptional induction of gene expression upon hypersalinity shock, followed by a negative feedback of gene expression. These results shed light on the regulation of glycerol synthesis during salt stress in Dunaliella. PMID- 19551476 TI - Tracing the factors responsible for arsenic enrichment in groundwater of the middle Gangetic Plain, India: a source identification perspective. AB - Arsenic contamination in groundwater is of increasing concern because of its high toxicity and widespread occurrence. This study is an effort to trace the factors responsible for arsenic enrichment in groundwater of the middle Gangetic Plain of India through major ion chemistry, arsenic speciation, sediment grain-size analyses, and multivariate statistical techniques. The study focuses on the distinction between the contributions of natural weathering and anthropogenic inputs of arsenic with its spatial distribution and seasonal variations in the plain of the state Bihar of India. Thirty-six groundwater and one sediment core samples were collected in the pre-monsoon and post-monsoon seasons. Various graphical plots and statistical analysis were carried out using chemical data to enable hydrochemical evaluation of the aquifer system based on the ionic constituents, water types, hydrochemical facies, and factors controlling groundwater quality. Results suggest that the groundwater is characterized by slightly alkaline pH with moderate to strong reducing nature. The general trend of various ions was found to be Ca(2+) > Na(+) > Mg(2+) > K(+) > NH(4) (+); and HCO(3) (-) > Cl(-) > SO(4) (2-) > NO(3) (-) > PO(4) (3-) > F(-) in both seasons. Spatial and temporal variations showed a slightly higher arsenic concentration in the pre-monsoon period (118 microg/L) than in the post-monsoon period (114 microg/L). Results of correlation analyses indicate that arsenic contamination is strongly associated with high concentrations of Fe, PO(4) (3-), and NH(4) (+) but relatively low Mn concentrations. Further, the enrichment of arsenic is more prevalent in the proximity of the Ganges River, indicating that fluvial input is the main source of arsenic. Grain size analyses of sediment core samples revealed clay (fine-grained) strata between 4.5 and 7.5 m deep that govern the vertical distribution of arsenic. The weathering of carbonate and silicate minerals along with surface-groundwater interactions, ion exchange, and anthropogenic activities seem to be the processes governing groundwater contamination, including with arsenic. Although the percentage of wells exceeding the permissible limit (50 microg/L) was less (47%) than that reported in Bangladesh and West Bengal, the percentage contribution of toxic As(III) to total arsenic concentration is quite high (66%). This study is vital considering that groundwater is the exclusive source of drinking water in the region and not only makes situation alarming but also calls for immediate attention. PMID- 19551477 TI - Comparative study of the nitrification characteristics of two different nitrifier immobilization methods. AB - The research investigated the nitrification characteristics of two different immobilization methods: nitrifier encapsulation in polyethylene glycol (PEG) gel pellets and nitrifier biofilm attachment on elastic plastic filler. The two carriers were placed in identical reactors. They reached a maximum nitrification rate of 39 and 25 mgN/L.h 30 days after start-up. The results showed that the nitrification efficiency in the PEG reactor was higher than in the biofilm reactor under the same conditions. Variations in temperature decreased the nitrification rate by approximately 55% in the PEG reactor from 28 to 8 degrees C, while 74.2% in the biofilm reactor. When the COD loading rate was increased to 0.8 kg/m(3) day, the nitrification efficiency in the biofilm reactor dropped sharply to 23%, and that of PEG reactor remained over 80%. PEG pellets with a high nitrification rate under all conditions showed promise as an immobilization medium, and are likely to be utilized in the nitrification of high-strength ammonia and COD wastewater during long-term operation. PMID- 19551478 TI - Expression of a putative stem cell marker, Musashi 1, in mammary glands of ewes. AB - Several recent studies demonstrated that development, function and remodelling of mammary glands involved multipotent cells, but no specific molecular markers for mammary epithelial stem cells were revealed. These studies principally concerned human and mouse mammary tissue, but mammary stem cells could be a valuable tool in agricultural production and bioengineering in farm animals. The Musashi-1 (Msi 1) gene encodes an RNA binding protein, which is likely to be associated with self-renewal of neural, intestinal and mammary progenitor cells and is believed to influence the Notch signalling pathway. In this study Musashi-1 expression was detected using immunohistochemistry and in situ hybridisation analysis on mammary glands of ewes at different developmental stages. The protein expression was observed in the epithelial cells at all stages examined. In situ hybridization analysis showed that Msi 1 mRNA has an expression pattern similar to the encoded protein, with positive staining in both nuclei and cytoplasm of ductal, secretory and stromal cells. Ultrastructural in situ analysis confirmed the nuclear and cytoplasmatic expression of Msi. Quantitative analysis of Msi 1 gene expression showed a strong correlation with that of Ki-67, that is a marker of cell proliferation. This is the first report outlining expression of Msi 1 in ovine mammary glands during a complete cycle of lactation. PMID- 19551479 TI - Thermal preconditioning prevents fiber type transformation of the unloading induced-atrophied muscle in rats. AB - Muscle atrophy is accompanied by a slow-to-fast transformation of the slow muscle, e.g., the soleus muscle, which is characterized by a decrease in the expression of the slow myosin heavy chain (MyHC) isoform. Heat stress before hindlimb unloading, i.e., thermal preconditioning, has been shown to reduce the rate of disuse-induced muscle atrophy. The present study examined whether thermal preconditioning could prevent a slow-to-fast transformation of the MyHC isoform through the induction of heat-shock protein (HSP) 72. Thermally preconditioned rats (Heat + HU) were individually placed in an environmentally controlled heat chamber for 1 h before hindlimb unloading for 2 weeks (HU). Although the mean fiber cross-sectional areas of the soleus muscle decreased in the HU and Heat + HU group, the loss of myofibrillar protein was attenuated in the Heat + HU group. Furthermore, a slow-to-fast transformation of MyHC isoform was inhibited in the Heat + HU group with the overexpression of HSP72. These results indicate that thermal preconditioning before hindlimb unloading attenuates the decrease of the slow MyHC isoform in the soleus muscle. Therefore, thermal preconditioning provides a new approach to prevent disuse-induced fiber type transformation of skeletal muscle. PMID- 19551481 TI - Bioassessment tools in novel habitats: an evaluation of indices and sampling methods in low-gradient streams in California. AB - Biomonitoring programs are often required to assess streams for which assessment tools have not been developed. For example, low-gradient streams (slopeor=450) to calculate the SCIBI in 28 of 45 samples and often collected fewer than 100 individuals, suggesting it is inappropriate for low-gradient streams in California; failures for the other methods were less common (TRC, 16 samples; MCM, 11 samples). Within-site precision, measured as the minimum detectable difference (MDD) was poor but similar across methods for the SCIBI (ranging from 19 to 22). However, RWB had the lowest MDD for O/E scores (0.20 versus 0.24 and 0.28 for MCM and TRC, respectively). Mantel correlations showed that assemblages were more similar within sites among methods than within methods among sites, suggesting that the sampling methods were collecting similar assemblages of organisms. Statistically significant disagreements among methods were not detected, although O/E scores were higher for RWB samples than TRC. Index scores suggested impairment at all sites in the study. Although index scores did not respond strongly to several measurements of disturbance in the watershed, percent agriculture showed a significant, negative relationship with O/E scores. PMID- 19551480 TI - Aeromonas and Pseudomonas: antibiotic and heavy metal resistance species from Iskenderun Bay, Turkey (northeast Mediterranean Sea). AB - We studied the susceptibility patterns to 15 different antibiotics and six heavy metals in Aeromonas spp. and Pseudomonas spp. isolated from Iskenderun Bay, Turkey (northeast Mediterranean Sea). A high percentage of Aeromonas isolates showed resistance to cefazolin (66.6%) and trimethoprim-sulphamethoxazole (66.6%). Amongst the Pseudomonas isolates, there was a high incidence of resistance to nitrofurantoin (86.2%), cefazolin (84.8%) and cefuroxime (71.7%). Most isolates showed tolerance to different concentrations of heavy metals, and minimal inhibition concentrations ranged from 25 to >3,200 microg/ml. The Aeromonas spp. and Pseudomonas spp. showed high resistance to copper of 98.3% and 75.4%, respectively, and low resistance to lead of 1.7% and 7.2%, respectively. Our results show that antibiotic and heavy metal resistant Aeromonas spp. and Pseudomonas spp. were widespread in Iskenderun Bay in 2007 and 2008. The increasing presence of antibiotic and heavy metal resistant Aeromonas spp. and Pseudomonas spp. may become a potential human health hazard. PMID- 19551482 TI - The anatomy of learning anatomy. AB - The experience of clinical teachers as well as research results about senior medical students' understanding of basic science concepts has much been debated. To gain a better understanding about how this knowledge-transformation is managed by medical students, this work aims at investigating their ways of setting about learning anatomy. Second-year medical students were interviewed with a focus on their approach to learning and their way of organizing their studies in anatomy. Phenomenographic analysis of the interviews was performed in 2007 to explore the complex field of learning anatomy. Subjects were found to hold conceptions of a dual notion of the field of anatomy and the interplay between details and wholes permeated their ways of studying with an obvious endeavor of understanding anatomy in terms of connectedness and meaning. The students' ways of approaching the learning task was characterized by three categories of description; the subjects experienced their anatomy studies as memorizing, contextualizing or experiencing. The study reveals aspects of learning anatomy indicating a deficit in meaningfulness. Variation in approach to learning and contextualization of anatomy are suggested as key-elements in how the students arrive at understanding. This should be acknowledged through careful variation of the integration of anatomy in future design of medical curricula. PMID- 19551483 TI - Occurrences of thermophilic Campylobacter in cattle slaughtered at Morogoro municipal abattoir, Tanzania. AB - An investigation was conducted in Morogoro municipality to assess the likelihood of slaughter cattle posing public health risk of contaminating carcasses with thermophilic Campylobacter. Butchers and meat shopkeepers were interviewed on source of slaughter cattle, method of animal and carcass transportation, carcass dressing, meat storage facilities, access to clean water and availability of food hygiene practices. Faecal samples were collected from 107 slaughter cattle and after slaughter; four different parts of dressed carcasses (i.e. from ham, neck, pelvis and thigh muscles) were also sampled. In addition 107 cattle meat samples for Campylobacter culture were collected in different meat shops. The samples were subjected to standard bacteriological examination using Skirrows protocol. It was found that cattle slaughter, dressing and meat handling in meatshops was done under unhygienic condition. Thermophilic Campylobacter prevalence in slaughter cattle was 5.6% while contamination rate of dressed carcasses and cattle meat at shops was 9.3% and 1.9%, respectively. The majority of thermophilic Campylobacter isolated were C. jejuni (88.9%) while C. coli was isolated at 11.1%. Findings of this study suggest possibility of humans acquiring zoonotic Campylobacter infections from cattle meat particularly when meat preparation and processing is not done properly. More work is required to establish the magnitude of zoonotic enteric Campylobacteriosis in humans and epidemiological role of cattle and other animals in the study area. PMID- 19551484 TI - Cross-sectional study on Contagious Caprine Pleuro Pneumonia in selected districts of sedentary and pastoral production systems in Southern Ethiopia. AB - A study to estimate the seroprevalence of Contagious Caprine Pluropneumonia (CCPP) in southern Ethiopia was conducted from November 2005 to June 2006. Two districts from sedentary (Arbaminch and Boreda) and pastoral (Hammar and Bena Tsemay) production systems were included in the study. Sera samples were collected from 913 goats (234 from sedentary and 679 from pastoral) to check for CCPP serostatus. The animals were sampled from 155 flocks (44 pastoral and 111 sedentary). Five clinically suspected CCPP cases were also sacrificed and attempt was made to isolate Mycoplasma capricolum capripneumoniae (MccP) from lung tissue, nasal swab and plural exudates. Sera samples were tested for the presence of CCPP antibodies using CFT. The overall seroprevalence recorded in the study was 18.61%. The corresponding seroprevalences for sedentary and pastoral production systems were 27.78% and 15.46% respectively. Regarding districts, the prevalence in Hammar was 15.63% while that of Bena-Tsemay 15.29%. In Arbaminch and Boreda the percent of seroreactors were 23.01 and 32.23% respectively. Out of 44 pastoral and 111 sedentary flocks, 50.45% of pastoral and 65.91% of sedentary flocks had at least one seroreactor goat per flock respectively. Both in the univariable and multivariable logistic regression analysis, seropositivity was found to have strong association with sedentary production system (P < 0.05, OR = 2.24) and adult age (P < 0.05, OR = 1.77). In microbiological study, two broth cultures from thoracic fluid and two broth cultures from lung tissue samples were found to be positive for Mycoplasma capricolum capripneumoniae (MccP). In conclusion, both the serological study and bacteriological isolation confirmed the disease CCPP being an important disease that demands serious attention in both production systems. PMID- 19551485 TI - The investigation of ureteral sympathetic innervation, using semi-serial sections: why does the alpha1-adrenergic receptor antagonist work well for ureteral stones? AB - PURPOSE: The efficacy of the alpha1-adrenergic receptor antagonist against ureteral stones was clarified on the basis of ureteral sympathetic innervation. METHODS: Semi-serial sections of retroperitoneal tissue were prepared from one each of donated male and female cadavers, and the nerve distribution in the ureter was investigated. Sympathetic nerve fibers were identified by immunostaining of tyrosine hydroxylase. RESULTS: We were able to prepare semi serial sections of the male right and female left ureters, in which 202 and 210 sympathetic nerve bundles were distributed. Sympathetic nerve bundles were distributed throughout the entire ureter. The number of nerve bundles was highest in the portion close to the bladder in the male right ureter (eight bundles/mm) and in the portion crossing the common iliac artery in the female left ureter (seven bundles/mm). CONCLUSION: Adrenergic sympathetic nerve fibers were distributed throughout the ureter. Therefore, alpha1-adrenergic receptor antagonists may act on the entire ureter, reducing its tonus. As a result, these antagonists may be useful for improving the stone-freeing rate and inhibiting pain attacks. PMID- 19551487 TI - Introduction to a special Festschrift issue celebrating the microbiology of Cupriavidus metallidurans strain CH34. PMID- 19551488 TI - Heart failure exacerbation leading to hospital admission: a cross-sectional study. AB - OBJECTIVES: The aim of this pilot study was to investigate cause(s) of heart failure (HF). SETTING: The emergency department and medical wards at Malmo University Hospital. METHOD: A cross sectional pilot study. MAIN OUTCOME MEASURES: Comparison of compliance, comprehension and optimal treatment on a population basis between men and women, younger (< or =75 years) and elderly (>75 years) patients, and patients in different New York Heart Association (NYHA) classes, in order to assess if exacerbation could have been caused by any of these factors. RESULTS: Of the 47 patients included, 60% reported high compliance, with significant differences between women and men, and between patients in NYHA class IV and patients in NYHA class III. Comprehension on self care was poor. Only 30% weighed themselves regularly and 45% did not limit the amount of fluids. No more than 28% reported they would contact a health professional in the case of experiencing more symptoms. Suboptimal treatment was also found to be a great concern. The majority were treated with recommended agents, but had not achieved target dose as recommended in the guidelines. CONCLUSION: This pilot study indicates suboptimal HF management of patients with HF prior to hospital admission due to HF exacerbation. A larger study is needed to assess the extent of the problem, and establish the need and nature of management improvement in different patient subgroups. PMID- 19551489 TI - Thermoluminescence: experimental. AB - Thermoluminesence measurements are useful for the study of Photosystem II electron transport in intact leaves, in algal and cyanobacterial cells, as well as in isolated membrane complexes. Here an overview of the experimental approaches is provided. In the present review, instruments and the experimental procedures for measuring thermoluminescence emission from photosynthetic systems of various origins are summarized and discussed. Major pitfalls frequently encountered in measurements with isolated membranes, suspensions of intact organisms or solid leaf samples are highlighted. Analytical and numeric methods for the analysis of measured thermoluminescence curves are also discussed. PMID- 19551490 TI - Disclosure and rationality: comparative risk information and decision-making about prevention. AB - With the growing focus on prevention in medicine, studies of how to describe risk have become increasing important. Recently, some researchers have argued against giving patients "comparative risk information," such as data about whether their baseline risk of developing a particular disease is above or below average. The concern is that giving patients this information will interfere with their consideration of more relevant data, such as the specific chance of getting the disease (the "personal risk"), the risk reduction the treatment provides, and any possible side effects. I explore this view and the theories of rationality that ground it, and I argue instead that comparative risk information can play a positive role in decision-making. The criticism of disclosing this sort of information to patients, I conclude, rests on a mistakenly narrow account of the goals of prevention and the nature of rational choice in medicine. PMID- 19551491 TI - Wisdom in clinical reasoning and medical practice. AB - Exploring informal components of clinical reasoning, we argue that they need to be understood via the analysis of professional wisdom. Wise decisions are needed where action or insight is vital, but neither everyday nor expert knowledge provides solutions. Wisdom combines experiential, intellectual, ethical, emotional and practical capacities; we contend that it is also more strongly social than is usually appreciated. But many accounts of reasoning specifically rule out such features as irrational. Seeking to illuminate how wisdom operates, we therefore build on Aristotle's work on informal reasoning. His account of rhetorical communication shows how non-formal components can play active parts in reasoning, retaining, or even enhancing its reasonableness. We extend this account, applying it to forms of healthcare-related reasoning which are characterised by the need for wise decision-making. We then go on to explore some of what clinical wise reasoning may mean, concluding with a case taken from psychotherapeutic practice. PMID- 19551492 TI - Yearning for certainty and the critique of medicine as "science". AB - A debate has simmered concerning the nature of clinical reasoning, especially diagnostic reasoning: is it a "science" or an "art"? The trend since the seventeenth century has been to regard medical reasoning as scientific reasoning, and the most advanced clinical reasoning is the most scientific. However, in recent years, several scholars have argued that clinical reasoning is clearly not "science" reasoning, but is in fact a species of narratival or hermeneutical reasoning. The study reviews this dispute, and argues that in a theoretical sense, the dispute rests upon a naive--but very popular--caricature of what constitutes "science reasoning." But, if the dispute rests upon just such a caricature, why is it so persistent? The study concludes by suggesting that we, as patients and as physicians, have deep psychological tendencies that incline us to adopt the very naive "science" concept/model of diagnostic reasoning, even if (or when) we understand its inaptness. PMID- 19551493 TI - Predictive testing for pre-malignancy as a prelude to adoption? An English case. AB - Genetic testing as a preliminary to adoption has been discussed in the literature [BAAF/BSHG Statement on the use of DNA testing to determine racial background], together with the medical benefits derived from such testing ASHG/ACMG Statement [Am J Hum Genetics, 66: 761-767, 2000]. But specific cases that reach court are rare. Such a case was recently discussed on national radio [http://www.bbc.co.uk/radio4/science/ethicscommittee_s4_tr3.shtml], disclosing sufficient facts to allow analysis of the ethical and legal issues encountered. Furthermore, the outcome of the case, a court order to test the child in infancy, can be reviewed in the light of the current law, together with the geneticist's response to this order. PMID- 19551495 TI - Barriers to seeking help for gambling problems: a review of the empirical literature. AB - This literature review summarizes recent empirical research on obstacles preventing problem gamblers from seeking treatment for their gambling problems. Relevant databases and bibliographies were searched for English-language papers and reports published since 1998. The only methodological requirement was that gamblers themselves be asked about reasons for not seeking help. Nineteen studies conducted in five countries were identified. All except one targeted adults. Despite differences in methodology, many of the same barriers to treatment were identified. Most commonly reported barriers were: wish to handle problem by oneself; shame/embarrassment/stigma; unwillingness to admit problem; and issues with treatment itself. The authors of the review argue that unwillingness to admit to the problem may be even more prevalent than is typically indicated by the results of barriers studies. Other frequently reported barriers included lack of knowledge about treatment options and practical issues around attending treatment. More research is needed on barriers to treatment-seeking experienced by subgroups of gamblers defined by culture, ethnicity, gender, age. Open-ended questioning methods can help provide insights into what barrier categories mean to different groups and individuals. Input directly from gamblers can be combined with information from other kinds of studies to devise better ways of reaching problem gamblers, especially those in underserviced populations. PMID- 19551496 TI - Prevalence of criminal thinking among state prison inmates with serious mental illness. AB - To examine the prevalence of criminal thinking in mentally disordered offenders, incarcerated male (n = 265) and female (n = 149) offenders completed measures of psychiatric functioning and criminal thinking. Results indicated 92% of the participants were diagnosed with a serious mental illness, and mentally disordered offenders produced criminal thinking scores on the Psychological Inventory of Criminal Thinking Styles (PICTS) and Criminal Sentiments Scale Modified (CSS-M) similar to that of non-mentally ill offenders. Collectively, results indicated the clinical presentation of mentally disordered offenders is similar to that of psychiatric patients and criminals. Implications are discussed with specific focus on the need for mental health professionals to treat co occurring issues of mental illness and criminality in correctional mental health treatment programs. PMID- 19551498 TI - STR polymorphisms of the Henan population and investigation of the Central Plains Han origin of Chaoshanese. AB - Allele frequencies for 15 short tandem repeat (STR) loci were obtained from a Chinese Han population in Henan province of middle China. No deviation from Hardy Weinberg equilibrium was observed for the STR loci except for D3S1358. The 15 STR loci are potentially useful for paternity testing and forensic casework in the Henan population. A phylogenetic tree based on CODIS STR allele frequencies of 25 Han populations revealed noticeable but far less clear distinctions between southern and northern Chinese populations; the Henan Han population was located at an intermediate position between south and north Chinese Han populations, relatively closer to Chaoshan and Minnan Han. Moreover, admixture analysis showed a large proportion of Central Plains Han origin in Chaoshanese and Minnanese. Admixture and phylogenetic analysis also reflected the genetic similarity shared by these two groups. PMID- 19551494 TI - LPS-induced cytokine levels are repressed by elevated expression of HSP70 in rats: possible role of NF-kappaB. AB - Heat shock protein (HSP)70 provides a spectrum of protection against any of a variety of stresses, preventing damage measured at the level of molecules, cells, as well as whole organism. We have previously reported that lipopolysaccharide (LPS)-induced lethality in rats is prevented by a previous exposure to a mild thermal stress and that a thermal stress sufficient to induce HSP70 expression in the liver is accompanied by an inhibition of endotoxin-mediated cytokines and modulation of febrile response. However, the effect of HSP70 upregulation on cytokine expression in animals is unknown. The aim of the present study was to demonstrate the effect of HSP70 overexpression with adenovirus administration on LPS-induced increase in cytokines levels in animals. In the present study, Sprague-Dawley rats were infected with either the control AdTrack or Ad70 virus that directs the expression of human HSP70. After a 5-day incubation, animals were injected with either saline alone or LPS (50 microg/kg). Four hours later, blood samples were drawn and plasma levels of interleukin (IL)-6 or tumor necrosis factor (TNF)-alpha were measured by enzyme-linked immunosorbent assay. Our data demonstrate for the first time that HSP70 overexpression with adenovirus injection prevented the LPS-induced increase in TNF-alpha and IL-6 levels in rats. Repression of LPS-induced cytokines expressions by HSP70 upregulation was associated with inhibited IkappaBalpha degradation and nuclear factor kappa-B (NF kappaB) p65 nuclear translocation in liver, suggesting that HSP70 overexpression may regulate LPS-induced cytokines expression through NF-kappaB pathway. We conclude that the effects of heat stress-induced increase in HSP70 protein expression on LPS-induced cytokine elaboration in whole animals can be reproduced by the actions of a single gene product. PMID- 19551499 TI - Predicting benefit from fulvestrant in pretreated metastatic breast cancer patients. AB - Fulvestrant use in pretreated metastatic breast cancer patients is associated with variable response rates. This study aimed to characterize these responses and to develop a prediction model to identify those patients who could potentially derive the most clinical benefit. A nationwide review of patients enrolled in a Canadian compassionate use program from 1999 to 2006 was performed. Prior therapy with tamoxifen, steroidal, and nonsteroidal aromatase inhibitors was mandatory. The dependent variable in the analysis was the proportion of patients requiring chemotherapy at 3 months following the start of fulvestrant. General Linear Mixed modeling was used to identify factors significantly associated with this dependant variable and to subsequently develop the prediction model. Three hundred and five women received at least one dose of fulvestrant; 207 went on to receive chemotherapy (68%). Median duration of fulvestrant treatment was 4.1 months (range 0.8-63.1). Factors predictive of being chemotherapy free at 3 months included older age, no prior adjuvant hormonal therapy, and the absence of lung or brain metastases at the start of therapy. A receiver operating characteristic (ROC) curve analysis had an area under the curve of 0.70 (95% CI 0.60-0.80). This model was able to identify risk information that could be helpful in assessing which patients would most likely benefit from fulvestrant as an intervention with the objective being a delay in chemotherapy. PMID- 19551500 TI - Validity of self-reports of breast cancer treatment in low-income, medically underserved women with breast cancer. AB - Few studies have assessed the agreement between subjects' self-report and medical records among patients with breast cancer (BC), and none has addressed this issue in low-income women with BC. We assessed the level of agreement between self report and medical records data for key BC treatment and prognostic characteristics using correct proportion and the Kappa statistic, among 726 low income BC patients. Unconditional regression was used to investigate the association between accuracy of self-report and potential explanatory factors. Overall agreement between self-report and medical records was 95.3-99.6% for BC treatments including surgery, chemotherapy, radiotherapy and hormone therapy (Kappa = 0.79-0.99). Specific agreement was 87-89.5% for surgery type (Kappa = 0.51-0.96); 86.3% for chemotherapy completion (Kappa = 0.46) and 98.7% for radiotherapy completion (Kappa = 0.43); 95.2% for medical oncologist consultation (Kappa = 0.59) and 96% for radiation oncologist consultation; 97.3% for metastasis (Kappa = 0.56); and 93.6% for recurrence (Kappa = 0.30). When accepting answers within 15 days of the medical record date, 78.2% of women correctly reported surgery date, yet only around 55% of women correctly reported the start and/or end date of radiotherapy. Older age, less education, BC recurrence and poor patient-physician communication were associated with the lesser accuracy of patients' self-report compared to medical records (P < 0.05). The results of this study suggest that self-reporting of key treatment and prognostic information is relatively accurate among low-income women with BC. Self-report seems to be a reliable source for accurate information when medical record review is unavailable or unfeasible. Interventions to enhance patient physician communication may facilitate more accurate information reporting among vulnerable populations. PMID- 19551501 TI - Comments on "Computer-aided detection in full-field digital mammography in a clinical population: performance of radiologist and technologists". PMID- 19551502 TI - An analysis of relationships among peer support, psychiatric hospitalization, and crisis stabilization. AB - This study's objective was to investigate how peer support relates to psychiatric hospitalization and crisis stabilization utilization outcomes. The likelihood of experiencing a psychiatric hospitalization or a crisis stabilization was modeled for consumers using peer support services and a control group of consumers using community mental health services but not peer support with 2003 and 2004 Georgia Medicaid claims data; 2003 and 2004 Mental Health, Developmental Disability, and Addictive Diseases (MHDDAD) Community Information System data; and 2003 and 2004 MHDDAD Hospital Information System data. Peer support was associated with an increased likelihood (odds = 1.345) of crisis stabilization, a decreased but statistically insignificant likelihood (odds = 0.871) of psychiatric hospitalization overall, and a decreased and statistically significant (odds = .766) likelihood of psychiatric hospitalization for those who did not have a crisis stabilization episode. PMID- 19551503 TI - Continuity of care: validation of a self-report measure to assess client perceptions of mental health service delivery. AB - The psychometric characteristics of an instrument to assess perceived continuity of care among mental health patients were examined. 441 adults with severe and persistent mental illness were recruited from 70 inpatient, outpatient, emergency and community treatment programs (n = 259 females; M age = 42.5, SD = 10.3 years) in Alberta, Canada. Respondents completed a 43-item self-report questionnaire to rate perceived continuity. Item quality was assessed by examining missing data and frequency distributions. In a randomly selected subsample (n = 171), exploratory factor analysis identified 3 dimensions: (1) perceived attentiveness to individual needs or changes in illness or life circumstances (Individualized Care; alpha = .64); (2) perceptions of a coherent system characterized by good communication between providers (Responsive System; alpha = .71); and (3) perceived responsiveness on the part of a primary provider (Responsive Caregiver; alpha = .52). In a second random subsample (n = 181), confirmatory factor analysis provided support for this 3-factor structure and the inclusion of a second-order "continuity" factor (alpha = .72), and informed further item reduction. Split-half reliability for the second-order factor was calculated. Associations between the factors and "objective" measures of continuity, as well as relevant clinical, quality of life, and service satisfaction variables, are reported. Use of the measure for clinical and research purposes and its limitations are considered. PMID- 19551504 TI - Field test of the feasibility and validity of using the Hoosier Assurance Plan Instrument for Adults in a state mental health program. AB - The current paper reports on the feasibility of using the HAPI-A, an instrument designed to assess a person's level of functioning in the community: (1) to help determine eligibility to receive behavioral health services, (2) to assign reimbursement case rates; and (3) to provide data for a service provider report card. A 3-year field study of the use of the instrument across an entire state mental health system explored the effectiveness of methods to enhance data accuracy, including annual training and a professional clinical record audit, and the ability of the test to detect differences in improvement rates within risk adjusted groupings. The combination of training and auditing produced statistically significant, cumulative reductions in data errors across all 3 years of the field test. The HAPI-A also was sensitive in detecting differences among service providers in outcome improvements for six of six risk-adjusted groups rated at the moderate level of impairment and for five of six groups rated at the mild level of impairment, but was inconsistent in detecting outcome differences for persons rated at the severe level of impairment. PMID- 19551505 TI - Patterns of recovery from severe mental illness: a pilot study of outcomes. AB - We performed a pilot study examining the patterns of recovery from severe mental illness in a model integrated service delivery system using measures from the Milestones of Recovery Scale (MORS), a valid and reliable measure of recovery outcomes which ranges from 1 to 8 (8 levels). For purposes of presentation, we constructed an aggregate MORS (6 levels) where the levels are described as follows: (1) extreme risk; (2) unengaged, poorly self-coordinating; (3) engaged, poorly self-coordinating; (4) coping and rehabilitating; (5) early recovery, and (6) self reliant. We analyzed MORS data on individuals followed over time from The Village in Long Beach, California (658 observations). Using Markov Chains, we estimated origin-destination transition probabilities, simulating recovery outcomes for 100 months. Our models suggest that after 12 months only 8% of "extreme risk" clients remain such. Over 40% have moved to "engaged, poorly self coordinating." After 2 years, almost half of the initial "extreme Risk" clients are "coping/rehabilitating", "early recovery" or "Self reliant." Most gains occur within 2 years. PMID- 19551506 TI - Barriers to and supports of family participation in a rural system of care for children with serious emotional problems. AB - Researchers have not adequately addressed the unique characteristics of rural areas that influence the accessibility of services for families with children who have serious emotional problems. Understanding rurality is particularly important to "systems of care" grant sites because these grants are intended to restructure mental health service delivery by building upon the strengths of a community and addressing the community's needs. This qualitative study examines the barriers to and supports for participation in services within a rural system of care site through the reported experiences of eight caregivers and nine staff. Findings indicate families face many challenges related to rurality, including stigma, transportation, isolation, poverty, and service availability. In addition to these challenges, however, participants reported many meaningful supports such as the religious community and the close-knit community of families and service providers. We present implications for planning, implementing, and evaluating systems of care in rural areas. PMID- 19551507 TI - Screening and access to services for individuals with serious mental illnesses in jails. AB - High rates of serious mental illnesses (SMI) among jail inmates pose challenges for the criminal justice system and risks for the individual with SMI. Research has identified actions to address these issues; it is unclear to what extent they have been operationalized. This study examines jails in one state, exploring how individuals with SMI are identified and treated, and comparing these with research-based recommendations. Results indicate that jails are not using evidence-based screenings, staff require training in SMI, access to services and medications for jailed individuals with SMI is often slow, and coordination between community providers and jails is limited. PMID- 19551508 TI - A simulation of vessel-clamp interaction: transient closure dynamics. AB - Cross-clamping of aorta is routinely performed in cardiac surgery. The objective of this study was to simulate cross-clamping of the aorta to elucidate the perturbation of stresses in the wall (solid mechanics) and lumen of the vessel (fluid mechanics). Models of the aorta and clamp were created in Computer Assisted Design and Finite Element Analysis packages. The vessel wall was considered as a non-linear anisotropic material while the fluid was simulated as Newtonian with pulsatile flow. The clamp was applied to produce total occlusion in approximately 1 s. A cylindrical and rectangular geometry for the clamp were considered. High jet speed and flow reversal were demonstrated during clamping. It was found that the clamp design and vessel wall anisotropy affected both the fluid wall shear stress (WSS) and solid stresses in vessel wall. The maximum wall stresses increased by about 170 and 220% during closure in the cases of plate and cylindrical clamps, respectively. The plate clamp design was superior for reduction of both solid stresses as well as fluid shear stresses. The cylindrical clamp causes much larger stresses than the plate clamp in each of the stress components; e.g., radial compression of -180 vs. -50 kPa. Vibrations, flow and WSS oscillations were detected immediately before total vessel occlusion. The present findings provide valuable insights into the mode of tissue injury during clamping and may also be useful for improving surgical clamp designs. PMID- 19551509 TI - Estimation of the biphasic property in a female's menstrual cycle from cutaneous temperature measured during sleep. AB - This paper proposes a method to estimate a woman's menstrual cycle based on the hidden Markov model (HMM). A tiny device was developed that attaches around the abdominal region to measure cutaneous temperature at 10-min intervals during sleep. The measured temperature data were encoded as a two-dimensional image (QR code, i.e., quick response code) and displayed in the LCD window of the device. A mobile phone captured the QR code image, decoded the information and transmitted the data to a database server. The collected data were analyzed by three steps to estimate the biphasic temperature property in a menstrual cycle. The key step was an HMM-based step between preprocessing and postprocessing. A discrete Markov model, with two hidden phases, was assumed to represent higher- and lower temperature phases during a menstrual cycle. The proposed method was verified by the data collected from 30 female participants, aged from 14 to 46, over six consecutive months. By comparing the estimated results with individual records from the participants, 71.6% of 190 menstrual cycles were correctly estimated. The sensitivity and positive predictability were 91.8 and 96.6%, respectively. This objective evaluation provides a promising approach for managing premenstrual syndrome and birth control. PMID- 19551510 TI - Role of upper airway dimensions in snore production: acoustical and perceptual findings. AB - While considerable efforts have been expended to develop snore-driven markers for detecting obstructive sleep apnea (OSA), there is little emphasis on the relationship between the human upper airway (UA) dimensions and the attributes of snores. This paper aims to investigate the acoustical and perceptual impacts of changing the cross-sectional areas (CSA) of the pharynx and oral cavity on the production of snores. Synthetic snores were generated based on the source-filter theory, whereas natural snores were recorded from 40 snorers during nocturnal polysomnography. First formant frequency (F1), spectral peak frequency (PF), and psychoacoustic metrics (loudness, sharpness, roughness, fluctuation strength, and annoyance) of CSA perturbations were examined, completed with diagnostic appraisal of F1 and PF for single- and mixed-gender groupings using the receiver operating characteristic curve analysis. Results show that (1) narrowing the pharyngeal airway consistently increases F1, but not for PF; and (2) altering the airway dimensions yield no considerable differences in perception of snore sounds, but indirectly affect the psychoacoustics by changing the dynamics of snore source flow. Diagnostic outcomes for all groupings (p-value < 0.0001) demonstrate that F1 is more capable of distinguishing apneic and benign snorers than PF due to the close association of F1 with the UA anatomical structures. Correlation exists between the UA anatomy and the properties of snores; there is a promising future for developing snore-driven screening tools for OSA. PMID- 19551511 TI - Investigation of obstructive sleep apnea using nonlinear mode interactions in nonstationary snore signals. AB - Acoustic studies on snoring sounds have recently drawn attention as a potential alternative to polysomnography in the diagnosis of obstructive sleep apnea (OSA). This paper investigates the feasibility of using nonlinear coupling between frequency modes in snore signals via wavelet bicoherence (WBC) analysis for screening of OSA. Two novel markers (PF1 and PSF), which are frequency modes with high nonlinear coupling strength in their respective WBC spectrum, are proposed to differentiate between apneic and benign snores in same- or both-gender snorers. Snoring sounds were recorded from 40 subjects (30 apneic and 10 benign) by a hanging microphone, and subsequently preprocessed within a wavelet transform domain. Forty inspiratory snores (30 as training and 10 as test data) from each subject were examined. Results demonstrate that nonlinear mode interactions in apneic snores are less self-coupled and usually occupy higher and wider frequency ranges than that of benign snores. PF1 and PSF are indicative of apneic and benign snores (p < 0.0001), with optimal thresholds of PF1 = 285 Hz and PSF = 492 Hz (for both genders combined), as well as sensitivity and specificity values between 85.0 and 90.7%, respectively, outperforming the conventional diagnostic indicator (spectral peak frequency, PF = 243-275 Hz, sensitivity = 77.7-79.7%, specificity = 72.0-78.0%, p < 0.0001). Relationships between apnea-hypopnea index and the proposed markers could likely take the functional form of exponential or power. Perspectives on nonlinear dynamics analysis of snore signals are promising for further research and development of a reliable and inexpensive diagnostic tool for OSA. PMID- 19551512 TI - Measurement of solute transport in the endothelial glycocalyx using indicator dilution techniques. AB - A new method is presented to quantify changes in permeability of the endothelial glycocalyx to small solutes and fluid flow using techniques of indicator dilution. Following infusion of a bolus of fluorescent solutes (either FITC or FITC conjugated Dextran70) into the rat mesenteric circulation, its transient dispersion through post-capillary venules was recorded and analyzed offline. To represent dispersion of solute as a function of radial position in a microvessel, a virtual transit time (VTT) was calculated from the first moment of fluorescence intensity-time curves. Computer simulations and subsequent in vivo measurements showed that the radial gradient of VTT within the glycocalyx layer (Delta VTT/Delta r) may be related to the hydraulic resistance within the layer along the axial direction in a post-capillary venule and the effective diffusion coefficient within the glycocalyx. Modeling the inflammatory process by superfusion of the mesentery with 10(-7) M fMLP, Delta VTT/Delta r was found to decrease significantly from 0.23 +/- 0.08 SD s/microm to 0.18 +/- 0.09 SD s/microm. Computer simulations demonstrated that Delta VTT/Delta r is principally determined by three independent variables: glycocalyx thickness (delta), hydraulic resistivity (K(r)) and effective diffusion coefficient of the solute (D(eff)) within the glycocalyx. Based upon these simulations, the measured 20% decrease in Delta VTT/Delta r at the endothelial cell surface corresponds to a 20% increase in D(eff) over a broad range in K(r), assuming a constant thickness delta. The absolute magnitude of D(eff) required to match Delta VTT/Delta r between in vivo measurements and simulations was found to be on the order of 2.5 x 10(-3) x D(free), where D(free) is the diffusion coefficient of FITC in aqueous media. Thus the present method may provide a useful tool for elucidating structural and molecular alterations in the glycocalyx as occur with ischemia, metabolic and inflammatory events. PMID- 19551513 TI - Design and fabrication of an injection tool for neuromuscular microstimulators. AB - The injection of small implants into precisely localized sites within the body is a difficult task usually undertaken by surgeons or interventive radiologists. We have designed, produced and tested a simple tool for implanting BION wireless microstimulators as an outpatient office procedure. The ability of BIONs to elicit a desired muscle contraction depends on their placement near the motor fibers that innervate the muscle fibers, providing both the requirement and a means for achieving accurate placement. The implant is preloaded into the tip of the cannula of a two-piece insertion tool made from non-conductive polymers. Trial stimulation pulses are generated by the implant as the tool is manipulated into the desired position. The implant is released by withdrawing the cannula over the implant, preserving both the relative location of the implant's electrodes with respect to the target and determining its desired axial orientation, which is important for implants containing motion sensors. The BION Insertion Tool has been used for over 30 BION implants in human subjects to date. PMID- 19551515 TI - A novel triple-regulated oncolytic adenovirus carrying PDCD5 gene exerts potent antitumor efficacy on common human leukemic cell lines. AB - PDCD5 (programmed cell death 5) accelerates apoptosis of certain tumor cells and the replication-defective Ad-PDCD5 may be a promising agent for enhancing chemosensitivity. In this study, a triple-regulated conditionally replicating adenoviruses (CRAd) carrying PDCD5 gene expression cassette, SG611-PDCD5, was engineered. In SG611-PDCD5, the E1a gene with a deletion of 24 nucleotides within CR2 region is controlled under the human telomerase reverse transcriptase (hTERT) promoter, the E1b gene expression is directed by the hypoxia response element (HRE), whereas the PDCD5 gene is controlled by the cytomegalovirus promoter. The tumor-selective replication of this virus and its antitumor efficacy were characterized in several leukemic cell lines in vitro and in xenograft models of human leukemic cell line in nude mice. It was found by RQ-RT-PCR assay that SG611 PDCD5 expressed PDCD5 efficiently in leukemic cells. In K562 tumor xenograft models, SG611-PDCD5 displayed a tumor killing capacity. At a dose of 1 x 10(9) plaque-forming units, SG611-PDCD5 alone could completely inhibit the tumor growth and more effective than replication-defective Ad-PDCD5. Histopathologic examination revealed that SG611-PDCD5 administration resulted in leukemic cell apoptosis. We concluded that the triple-regulated SG611-PDCD5, as a more potent and safer antitumor therapeutic, could provide a new strategy for leukemia biotherapy. PMID- 19551517 TI - A two one-sided parametric tolerance interval test for control of delivered dose uniformity--part 3--investigation of robustness to deviations from normality. AB - The robustness of the parametric tolerance interval test, which was proposed by the Food and Drug Administration for control of delivered dose uniformity in orally inhaled and nasal drug products, is investigated in this article using different scenarios for deviations from a univariate normal distribution. The studied scenarios span a wide range of conditions, the purpose of which is to provide an understanding of how the test performs depending on the nature and degree of the deviation from normality. Operating characteristic curves were generated to compare the performance of the test for different types of distributions (normal and non-normal) having the same proportion of doses in the tails (on one or both sides) outside the target interval. The results show that, in most cases, non-normality does not increase the probability of accepting a batch of unacceptable quality (i.e., the test is robust) except in extreme situations, which do not necessarily represent commercially viable products. The results also demonstrate that, in the case of bimodal distributions where the life-stage means differ from each other by up to 24% label claim, the test's criterion on life-stage means does not affect pass rates because the tolerance interval portion of the test reacts to shifting means as well. PMID- 19551516 TI - Collision sellar lesions: experience with eight cases and review of the literature. AB - The concomitant presence of a pituitary adenoma with a second sellar lesion in patients operated upon for pituitary adenoma is an uncommon entity. Although rare, quite a great variety of lesions have been indentified coexisting with pituitary adenomas. In fact, most combinations have been described before, but an overview with information on the frequency of combined pathologies in a large series has not been published. We present a series of eight collision sellar lesions indentified among 548 transsphenoidally resected pituitary adenomas in two Neurosurgical Departments. The histological studies confirmed a case of sarcoidosis within a non-functioning pituitary adenoma, a case of intrasellar schwannoma coexisting with growth hormone (GH) secreting adenoma, two Rathke's cleft cysts combined with pituitary adenomas, three gangliocytomas associated with GH-secreting adenomas, and a case of a double pituitary adenoma. The pertinent literature is discussed with emphasis on pathogenetic theories of dual sellar lesions. Although there is no direct evidence to confirm the pathogenetic relationship of collision sellar lesions, the number of cases presented in literature makes the theory of an incidental occurrence rather doubtful. Suggested hypotheses about a common embryonic origin or a potential interaction between pituitary adenomas and the immune system are presented. PMID- 19551518 TI - A two one-sided parametric tolerance interval test for control of delivered dose uniformity. Part 1--characterization of FDA proposed test. AB - The FDA proposed a parametric tolerance interval (PTI) test at the October 2005 Advisory Committee meeting as a replacement of the attribute (counting) test for delivered dose uniformity (DDU), published in the 1998 draft guidance for metered dose inhalers (MDIs) and dry powder inhalers (DPIs) and the 2002 final guidance for inhalation sprays and intranasal products. This article (first in a series of three) focuses on the test named by the FDA "87.5% coverage." Unlike a typical two-sided PTI test, which controls the proportion of the DDU distribution within a target interval (coverage), this test is comprised of two one-sided tests (TOST) designed to control the maximum amount of DDU values in either tail of the distribution above and below the target interval. Through simulations, this article characterizes the properties and performance of the proposed PTI-TOST under different scenarios. The results show that coverages of 99% or greater are needed for a batch to have acceptance probability 98% or greater with the test named by the FDA "87.5% coverage" (95% confidence level), while batches with 87.5% coverage have less than 1% probability of being accepted. The results also illustrate that with this PTI-TOST, the coverage requirement for a given acceptance probability increases as the batch mean deviates from target. The accompanying articles study the effects of changing test parameters and the test robustness to deviations from normality. PMID- 19551519 TI - A two one-sided parametric tolerance interval test for control of delivered dose uniformity--part 2--effect of changing parameters. AB - This article examines the effects of changing parameters in the test which was proposed by the FDA at the October 2005 Advisory Committee meeting for confirming delivered dose uniformity in orally inhaled and nasal drug products. This article is an extension of the characterization study presented in an accompanying article (Part 1). The goal of this study is to understand how parameters of the test affect the test performance. The effects of changing test parameters such as target interval, maximum allowable proportion in the tail area, and sample size are examined. The results show that changing the maximum allowable tail area and/or the target interval have the largest impact on the test outcomes, i.e., probability of acceptance for a given batch mean and standard deviation. The presented information may provide potential users of the test with a set of tools for optimizing the test characteristics for a particular product. PMID- 19551520 TI - Challenges for the autoimmunologist. AB - Every 2 years, an International Congress of Autoimmunity is organized and held with the intent of creating a new base for continuing interdisciplinary and international research, including further opportunities to train in the field of autoimmunity. In September 2008, the Congress of Autoimmunity was held in Porto, Portugal, and over the next 6 months, requests were made from a number of key investigators to provide copies of their papers for publication in a special volume of this journal. In addition, a satellite meeting sponsored by the AESKU.KIPP Institute was held in Porto with an attempt of focusing on specific challenges faced by the autoimmunologist. It is hoped that this collection of papers will become a valuable source for researchers in autoimmunity. PMID- 19551521 TI - Influence of insulin treatment on the lacrimal gland and ocular surface of diabetic rats. AB - Previous studies have observed changes in the lacrimal gland and ocular surface related to diabetes mellitus and related it to insulin resistance or insufficiency and oxidative damage. The aim of this study was to evaluate whether insulin treatment inhibits those changes. Diabetes was induced in male Wistar rats with a single intravenous injection of streptozotocin and a subgroup was treated with insulin. After 5 and 10 weeks, the three groups (n = 5 10/group/experimental procedure) were compared for biochemical, functional, and histological parameters. After 5 weeks, changes in morphology and increased numbers of lipofucsin-like inclusions were observed in lacrimal glands of diabetic but not insulin-treated rats. After 5 weeks, malonaldehyde and total peroxidase activity were significantly higher in diabetic rats, but similar to control in insulin-treated diabetic rats (P = 0.03, P = 0.02, respectively). Our data indicate that diabetes induces histological alterations in lacrimal gland and suggests that hyperglycemia-related oxidative stress may participate in diabetic dry eye syndrome. Prevention by insulin replacement suggests direct hormone action and/or benefit by early sub optimal metabolic control. PMID- 19551522 TI - Effects of estrogen receptor alpha and beta gene deletion on estrogenic induction of progesterone receptors in the locus coeruleus in female mice. AB - Locus coeruleus (LC) is involved in the LHRH regulation by gonadal steroids. We investigated the expression of progesterone and estrogen receptors (PR; ER) in LC neurons of ERalpha (alphaERKO) or ERbeta (betaERKO) knockout mice, and their wild type (alphaWT and betaWT). Immunocytochemical studies showed that LC expresses PR and both ERs, although ERbeta was more abundant. Estradiol benzoate (EB) decreased ERalpha-positive cells in WT and betaERKO mice, and progesterone caused a further reduction, whereas none of the steroids influenced ERbeta expression. ERbeta deletion increased ERalpha while ERalpha deletion did not alter ERbeta expression. In both WT mice, EB increased PR expression, which was diminished by progesterone. These steroid effects were also observed in alphaERKO animals but to a lesser extent, suggesting that ERalpha is partially responsible for the estrogenic induction of PR in LC. Steroid effects on PR in betaERKO mice were similar to those in the alphaERKO but to a lesser extent, probably because PR expression was already high in the oil-treated group. This expression seems to be specific of LC neurons, since it was not observed in other areas studied, the preoptic area and ventromedial nucleus of hypothalamus. These findings show that LC in mice expresses alphaER, betaER, and PR, and that a balance between them may be critical for the physiological control of reproductive function. PMID- 19551523 TI - Targeting myofibroblasts in model systems of fibrosis by an artificial alpha smooth muscle-actin promoter hybrid. AB - Myofibroblasts are the main cell types producing extracellular matrix proteins in a variety of fibrotic diseases. Therefore, they are useful targets for studies of intracellular communication and gene therapeutical approaches in scarring diseases. An artificial promoter containing the -702 bp regulatory sequence of the alpha-smooth muscle actin (SMA) gene linked to the first intron enhancer sequence of the beta-actin gene and the beta-globin intron-exon junction was constructed and tested for myofibroblast-dependent gene expression using the green fluorescent protein as a reporter. Reporter expression revealed myofibroblast-specific function in hepatic and renal myofibroblasts, in vitro. In addition, differentiation-dependent activation of the SMA-beta-actin promoter hybrid was shown after induction of myofibroblastic features in mesangial cells by stretching treatment. Furthermore, wound healing experiments with SMA-beta actin promoter reporter mice demonstrated myofibroblast-specific action, in vivo. In conclusion, the -702 bp regulatory region of the SMA promoter linked to enhancing beta-actin and beta-globin sequences benefits from its small size and is suggested as a promising tool to target myofibroblasts as the crucial cell type in various scarring processes. PMID- 19551524 TI - The high mobility group protein HMGA2: a co-regulator of chromatin structure and pluripotency in stem cells? AB - The small, chromatin-associated HMGA proteins contain three separate DNA binding domains, so-called AT hooks, which bind preferentially to short AT-rich sequences. These proteins are abundant in pluripotent embryonic stem (ES) cells and most malignant human tumors, but are not detectable in normal somatic cells. They act both as activator and repressor of gene expression, and most likely facilitate DNA architectural changes during formation of specialized nucleoprotein structures at selected promoter regions. For example, HMGA2 is involved in transcriptional activation of certain cell proliferation genes, which likely contributes to its well-established oncogenic potential during tumor formation. However, surprisingly little is known about how HMGA proteins bind DNA packaged in chromatin and how this affects the chromatin structure at a larger scale. Experimental evidence suggests that HMGA2 competes with binding of histone H1 in the chromatin fiber. This could substantially alter chromatin domain structures in ES cells and contribute to the activation of certain transcription networks. HMGA2 also seems capable of recruiting enzymes directly involved in histone modifications to trigger gene expression. Furthermore, it was shown that multiple HMGA2 molecules bind stably to a single nucleosome core particle whose structure is known. How these features of HMGA2 impinge on chromatin organization inside a living cell is unknown. In this commentary, we propose that HMGA2, through the action of three independent DNA binding domains, substantially contributes to the plasticity of ES cell chromatin and is involved in the maintenance of a un-differentiated cell state. PMID- 19551526 TI - Hematological indices at birth in relation to arterial and venous Doppler in small-for-gestational-age fetuses. AB - OBJECTIVE: To correlate Doppler results with hematological indices at birth in small-for-gestational-age (SGA) fetuses. DESIGN: Prospective study. SETTING: Tertiary teaching hospital, Sao Paulo, Brazil. Population. One hundred singleton pregnancies with SGA fetuses of > 27 weeks gestational age. METHODS: All women had Doppler velocimetry of the umbilical arteries, middle cerebral artery, and ductus venosus within < 72 hours prior to delivery. After birth, umbilical artery blood was collected for hematological analysis. MAIN OUTCOME MEASURES: The association between fetal Doppler velocimetry pulsatility index (PI) and some hematological indices. RESULTS: Umbilical artery PI showed a positive correlation with nucleated red blood cell count in the umbilical cord (r = 0.46; p < 0.01), and a negative correlation with platelet count (r = -0.53; p < 0.01) and white blood cell count (r = -0.42; p < 0.01). Middle cerebral artery PI was positively correlated with platelet count (r = 0.43; p < 0.01) and white blood cell count (r = 0.38; p < 0.01), and was negatively correlated with nucleated red blood cell count (r = -0.39; p < 0.01). The ductus venosus pulsatility index showed a positive correlation with nucleated red blood cell count (r = 0.36; p < 0.01), and a negative correlation with platelet count (r = -0.37; p < 0.01) and white blood cell count (r = -0.26; p < 0.01). CONCLUSION: A significant positive or negative correlation between nucleated red blood cell, platelet and white blood cell counts, and Doppler indices suggests an association between placental insufficiency and the fetal hematological response. PMID- 19551527 TI - Peripheral nerves are sensitive to irradiation, Or: on the pitfalls from too short follow-up times. PMID- 19551528 TI - Social ties and risk for cancer--a prospective cohort study. AB - BACKGROUND: Poor social support and small social networks have been associated with increased risks for conditions such as coronary heart disease as well as with overall mortality. We investigated the association between social ties and risk for cancer. MATERIAL AND METHODS: The study sample consisted of 8 548 Danes who had been examined in 1991-1994 within the Copenhagen City Heart Study. The median length of follow-up was 9.3 years (range, 0-11.2 years). Social ties were measured from answers to a questionnaire on social networks. Regression analyses for cancers at the most frequent sites (breast, lung, prostate and colon and rectum) were conducted with the Cox proportional hazards model, with adjustment for a number of well-known risk factors for cancer. RESULTS: While we found no significant association between social ties and risk for cancer in men, women with high social network scores had an increased risk for lung cancer of borderline significance (HR, 2.16; 95% CI, 1.02-4.60). The risks for breast cancer and colorectal cancers were not significantly increased in the same group of women. DISCUSSION: The results of this study do not support the hypothesis that social network size is associated with a decreased risk for cancer. PMID- 19551525 TI - The molecular mechanism of induced pluripotency: a two-stage switch. AB - Pluripotent stem cells are basic cells with an indefinite self-renewal capacity and the potential to generate all the cell types of the three germinal layers. So far, the major source for pluripotent stem cells is the inner cell mass of the blastocysts: embryonic stem (ES) cells. Potential clinical application of ES cells is faced with many practical and ethical concerns. So, a major breakthrough was achieved in 2006, when it was shown that pluripotent stem cells could be obtained by transducing mouse embryonic and adult fibroblasts with a limited set of defined transcription factors. These reprogrammed cells, named induced pluripotent stem (iPS) cells, resembled ES cells in many of their characteristics. Since this initial study, iPS cell research has taken an incredible flight, and to date iPS cells have been generated from cells from several species using different sets of reprogramming factors. Given the potential to generate patient-specific cell populations without the need for human embryonic cells, iPS cell technology has been received with great excitement by research and medical communities. However, many questions regarding the actual molecular process of induced reprogramming remain unanswered and need to be addressed before iPS cells can go to the clinic. In this review, we start by summarizing recent advances in iPS cell research and inventory the hurdles that still need to be taken before safe clinical application. Our major aim, however, is to review the available data on the molecular processes underlying pluripotency reprogramming and present a two-stage switch model. PMID- 19551530 TI - Incidence and survival of gastric non-Hodgkin's lymphoma: a population-based study from the Association of the French Cancer Registries (FRANCIM). AB - BACKGROUND: Most epidemiological studies on gastric lymphomas (GL) were carried out before changes in therapy were introduced. The aim of the study was to measure the incidence of GL and to estimate survival. MATERIAL AND METHODS: Data were provided by the Association of the French Cancer Registries database. Age standardized incidence rates were calculated for 786 incident cases diagnosed between 1978 and 2002. Crude and relative survival were calculated for 361 cases diagnosed between 1989 and 1997. Effects specific to sex, age at diagnosis, year of diagnosis, and grade of malignancy were estimated in multivariate analysis. RESULTS: Incidence was stable during the study period. However, high-grade GL frequency increased whereas low-grade and not otherwise specified (NOS) GL frequencies were respectively stable and decreased. At 5 years, relative survival was 63% in men and 60% in women. Patients aged 75 or older had a five-year relative survival of 33%. Age at diagnosis was the only significant prognostic factor in multivariate analysis. Time trend improvement in prognosis was observed. DISCUSSION: Results in elderly patients show that therapeutic regimens should be specifically designed and assessed for them. The prognosis improvement trend is probably related to the implementation of changes in management of patients and has to be confirmed by more recent data. PMID- 19551532 TI - Somatic mutation of GNAQ gene is rare in common solid cancers and leukemias. PMID- 19551533 TI - Correction for patient sway in radiographic biplanar imaging for three dimensional reconstruction of the spine: in vitro study of a new method. AB - BACKGROUND: Three-dimensional (3D) reconstructions of the spine in the upright position are classically obtained using two-dimensional, non-simultaneous radiographic imaging. However, a subject's sway between exposures induces inaccuracy in the 3D reconstructions. PURPOSE: To evaluate the impact of patient sway between successive radiographic exposures, and to test if 3D reconstruction accuracy can be improved by a corrective method with simultaneous Moire-X-ray imaging. MATERIAL AND METHODS: Using a calibrated deformable phantom perceptible by both techniques (Moire and X-ray), the 3D positional and rotational vertebral data from 3D reconstructions with and without the corrective procedure were compared to the corresponding data of computed tomography (CT) scans, considered as a reference. All were expressed in the global axis system, as defined by the Scoliosis Research Society. RESULTS: When a sagittal sway of 10 degrees occurred between successive biplanar X-rays, the accuracy of the 3D reconstruction without correction was 8.8 mm for the anteroposterior vertebral locations and 6.4 degrees for the sagittal orientations. When the corrective method was applied, the accuracy was improved to 1.3 mm and 1.5 degrees , respectively. CONCLUSION: 3D accuracy improved significantly by using the corrective method, whatever the subject's sway. This technique is reliable for clinical appraisal of the spine, if the subject's sway does not exceed 10 degrees . For greater sway, improvement persists, but a risk of lack of accuracy exists. PMID- 19551534 TI - Fractures of self-expanding metallic stents in periampullary malignant biliary obstruction. AB - BACKGROUND: Self-expanding metallic stents are widely used for relieving biliary duct obstruction in patients with unresectable periampullary malignancies. However, only a few studies have assessed the occurrence of fractures in these stents. PURPOSE: To determine the prevalence and significance of stent fracture after placement of self-expanding metallic stents for periampullary malignant biliary obstruction. MATERIAL AND METHODS: Over a 5-year period, 48 patients underwent placement of self-expanding metallic stents for periampullary malignant biliary obstructions. Stents were introduced 2-6 weeks after a percutaneous transhepatic biliary decompression. The medical records and relevant images were reviewed for stent patency, stent fracture, type of stent, and stent-related complications. RESULTS: Stent fracture was detected in four of the 48 patients (8%): in one patient at 1 month and in three patients between 10 and 21 months after stenting. All four fractures involved one type of nitinol stent used in 38 patients. In one of the patients, fracture was complicated by life-threatening gastrointestinal bleeding. The mean survival time for all patients was 251 days (standard deviation [SD]+/-275 days) and the mean overall patency time for all stents was 187 days (SD+/-205 days). CONCLUSION: Stent fracture occurs after placement of self-expanding nitinol stents for periampullary malignant biliary obstruction. The low reported incidence of this complication may be due to a lack of awareness of and difficulty in detecting stent fracture. Fracture should be considered as a possible contributing factor in recurrent biliary obstruction after self-expanding metallic stent insertion. PMID- 19551535 TI - Homocysteine levels and amyotrophic lateral sclerosis: A possible link. AB - Homocysteine (Hcy) exerts multiple neurotoxic mechanisms that have also been shown to be relevant in the pathogenesis of amyotrophic lateral sclerosis (ALS). We reviewed the published evidence to assess possible correlations between Hcy and ALS. A Medline literature search was performed to identify all studies on Hcy and ALS or motor neurons published from 1 January 1966 through 28 February 2009. Twelve studies (one in vitro, eight in vivo, and three studies on human subjects) were reviewed. The in vitro and in vivo animal studies showed that Hcy can damage motor neurons by inducing oxidative stress and stimulating excitotoxic receptors. In preliminary studies on human subjects, ALS subjects had higher median Hcy levels compared to age- and sex-matched controls. Higher Hcy levels were also correlated with a possible marker of disease progression. Finally, a short-term treatment with a high dose of methylcobalamin, which reduces Hcy levels, was effective in improving compound motor action potentials in patients with ALS. In conclusion, several types of evidence show that accumulation of Hcy may increase the risk and progression of motoneuronal degeneration. If this is confirmed, early interventions to decrease Hcy levels may be useful to modify ALS progression and possibly onset. PMID- 19551536 TI - Correlates of quality of life in ALS: Lessons from the minocycline study. AB - Improving quality of life (QoL) is a major goal in ALS palliative care. Previous studies performed on the general ALS population showed no relationship between QoL and disease progression. ALS subjects participating in clinical trials may differ from those in the general ALS population. We explored the relationship between QoL and disease progression in 412 subjects enrolled in a minocycline trial. We examined correlations between Single Item McGill Quality of Life Scale (MQoL-SIS) score and disease duration, ALS Functional Rating Scale Revised (ALSFRS-R) score, FVC, and survival rate. We also analyzed how NIV and PEG affect QoL. Within subjects, MQoL-SIS scores correlated with ALSFRS-R and FVC (p<0.001). MQoL-SIS declined over time (p<0.001) and correlated with the decline of ALSFRS-R (p<0.001). MQoL-SIS tended to improve after initiation of NIV (p=0.07). There was a significant reduction in the rate of MQoL-SIS decline (p<0.001) after initiation of PEG. Subjects with slower QoL decline survived seven months longer than those with faster QoL decline (p<0.01). Our study demonstrated that QoL does decline with advancing ALS in subjects who participated in a minocycline trial, that the slope of QoL predicts survival, and that both NIV and PEG have beneficial impacts on QoL. PMID- 19551537 TI - Prognosis of unrecognized coeliac disease as regards mortality: a population based cohort study. AB - BACKGROUND AND AIM: Clinically diagnosed coeliac disease patients carry an increased risk of mortality. As coeliac disease is markedly underdiagnosed, we aimed to quantify the risk of mortality in subjects with unrecognized and thus untreated coeliac disease. METHOD: Blood samples from 6,987 Finnish adults were drawn in 1978-80, and sera were tested for immunoglobulin A (IgA)-class tissue transglutaminase antibodies (Eu-tTG) in 2001. Positive sera were further analysed for endomysial (EMA) and tissue transglutaminase antibodies by another test (Celikey tTG). EMA- and Celikey tTG-positive cases were compared to negatives as regards mortality in up to 28 years of surveillance, yielding a total follow-up of 147,646 person years. Dates and causes of death were extracted from the nation wide database. RESULTS: Altogether 74 (1.1%) of the participants were EMA- and 204 (2.9%) Celikey tTG-positive. The age- and sex-adjusted relative risk of overall mortality was not increased in either EMA (0.78, 95% CI 0.52-1.18) or Celikey tTG (1.19, 95% CI 0.99-1.42) -positive subjects. However, antibody positive cases evinced a tendency to die from lymphoma, stroke, and diseases of the respiratory system. CONCLUSIONS: The prognosis of unrecognized coeliac disease was good as regards overall mortality, which does not support screening of asymptomatic coeliac disease cases. PMID- 19551538 TI - Depressive symptoms in the congenital long QT syndrome. AB - BACKGROUND: A proportion of patients with congenital long QT syndrome (LQTS) experience potentially life-threatening cardiac arrhythmias. AIM: To examine whether depressive symptoms are related to arrhythmic events among symptomatic and asymptomatic LQTS patients, and syncope events among their relatives not carrying the family's LQTS-causing mutation. METHODS: The participants were 569 molecularly defined LQTS mutation carriers and 622 non-carrier relatives from the Finnish LQTS registry. Depressive symptoms were self-rated with a revised version of the Beck Depression Inventory. RESULTS: LQTS patients with arrhythmic events scored higher on depressive symptoms than those without (P=0.011) or the control group (P=0.005). In addition, in the binary logistic regression analysis including symptomatic and asymptomatic LQTS mutation carriers, depressive symptoms showed an age- and sex-adjusted association of odds ratio (OR) 1.40 (95% confidence interval (CI) 1.12-1.74) with symptomatic status of LQTS. In similar analysis including non-carriers of the LQTS mutation, there was no association between depressive symptoms and history of syncope events OR 1.23 (95% CI 0.99 1.53). CONCLUSION: Our results from this relatively large genotyped LQTS patient cohort indicate that depressive symptoms are associated with arrhythmic events in LQTS patients. Whether depressive symptoms are causally related to arrhythmias in LQTS remains uncertain. PMID- 19551540 TI - Sex and sexuality in the Shenzhen tongzhi circle: HIV risk context and migrant men who have sex with men in China. AB - The paper explores the HIV risk context of migrant men who have sex with men, or tongzhi, in Shenzhen, China. Findings from 42 qualitative interviews indicate that respondents are living within a complex risk environment, characterised by the realities of migration and a 'weak ties', virtual men who have sex with men community, which offers little emotional support but many opportunities for sexual engagement. Male sex work has proliferated in Shenzhen, with respondents informally participating in a type of buy-sell circuit. Respondents' portrayal of their environment and conflicts were underscored by three interacting elements: sexual freedom, migration and traditional Chinese values. The confluence of these factors, and how they may influence risk prioritisation in a multi-risk environment, is explored. PMID- 19551544 TI - Efficacy of myo-inositol in the treatment of cutaneous disorders in young women with polycystic ovary syndrome. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is the most common endocrine cause of hirsutism, acne and pattern alopecia, often characterised by ovulation disorders (usually manifested as oligo- or amenorrhea). In addition, 30-40% of women with PCOS have impaired glucose tolerance, and a defect in the insulin signalling pathway seems to be implicated in the pathogenesis of insulin resistance. For this reason, insulin-lowering medications represent novel approach in women with PCOS. The aim of this study was to evaluate the effects of myo-inositol (MYO), an isoform of inositol, belonging to the vitamin B complex, in the treatment of cutaneous disorders like hirsutism and acne. METHODS: Fifty patients with PCOS were enrolled in the study. BMI, LH, FSH, insulin, HOMA index, androstenedione, testosterone, free testosterone, hirsutism and acne were evaluated at the baseline and after receiving MYO therapy for 6 months. RESULTS: After 3 months of MYO administration, plasma LH, testosterone, free testosterone, insulin and HOMA index resulted significantly reduced; no significant changes were observed in plasma FSH and androstenedione levels. Both hirsutism and acne decreased after 6 months of therapy. DISCUSSION: MYO administration is a simple and safe treatment that ameliorates the metabolic profile of patients with PCOS, reducing hirsutism and acne. PMID- 19551542 TI - In vitro study of matrix metalloproteinase/tissue inhibitor of metalloproteinase production by mesenchymal stromal cells in response to inflammatory cytokines: the role of their migration in injured tissues. AB - BACKGROUND AIMS: The transmigratory capacity of bone marrow (BM) mesenchymal stromal cells (MSC) through the endothelial cell barrier into various tissues and their differentiation potential makes them ideal candidates for cell therapy. Nevertheless, the mechanisms and agents promoting their migration are not fully understood. We evaluated the effects of several inflammatory cytokines on the migration of BM MSC and matrix metalloproteinase (MMP)/tissue inhibitor of metalloproteinase (TIMP) production. METHODS: The migratory potential of BM MSC was evaluated using a Boyden chamber coated with Matrigel in the presence and absence of stromal cell-derived (SDF)-1alpha, platelet-derived growth factor (PDGF)bb, insulin-like growth factor (IGF)-I and interleukin (IL)-6. The ability of inflammatory cytokines to induce MSC migration was tested in presence of their respective Ab or blocking peptide. We used immunofluorescence to check the expression of cytokine receptors, and MMP/TIMP production was analyzed at the protein (human cytokine array, enzyme-linked immunosorbent assay (ELISA), gelatine zymography and Western blot) and mRNA quantitative real-time polymerase chain reaction (qRT-PCR) levels. RESULTS: We have demonstrated that inflammatory cytokines promote the migratory capacity of BM MSC according to the expression of their respective receptors. Higher migration through Matrigel was observed in response to IL-6 and PDGFbb. qRT-PCR and cytokine array revealed that migration was the result of the variable level of MMP/TIMP in response to inflammatory stimuli. CONCLUSIONS: Our observations suggest that chemokines and cytokines involved in the regulation of the immunity or inflammatory process promote the migration of MSC into BM or damaged tissues. One of the mechanisms used by MSC to promote their migration though the extracellular matrix is modulation of the production of MMP-1, MMP-2, MMP-13, TIMP-1 and TIMP-2. PMID- 19551545 TI - Heat shock proteins in antigen trafficking--implications on antigen presentation to T cells. AB - Heat shock proteins (HSP) are molecular chaperones implicated in facilitation of protein folding and translocation between distinct compartments, and hence in preventing protein from aggregation. In terms of proteolysis, HSP act as a double edged sword, stimulating proteasome-dependent proteolysis while preventing the degradation of the same proteins, even though in both cases association of unfolded proteins with HSP is the initial step. The proteasomal degradation products are utilised as ligands of major histocompatibility complex (MHC) class I molecules to be recognised by CD8(+) T cells, leading to activation of cytotoxic T cell immunity indispensable in fighting virus infections and cancers. In this context, HSP-mediated antigen traffic towards proteasomal degradation is coupled with acquired T cell immunity. In addition, exogenous antigens internalised by dendritic cells (DC) are also forwarded to the proteasome, possibly through the ER-associated degradation (ERAD) system, based on the fusion of the ER-membrane to the endosome containing the antigens. Thus, antigens within endosomes might be translocated to the cytosol, possibly through the Sec61 complex recruited from ER and degraded by the proteasome, rendering their peptides presentable by MHC class I molecules, a process known as cross presentation. Since binding protein (Bip) facilitates degradation of most ER luminal soluble proteins in yeast, it is possible that endosomal HSP in DC, mimicking the action of Bip, facilitate the degradation of internalised soluble antigens. This may explain why the HSP-peptide/protein complex is extremely efficient in terms of cross-presentation ability. In this review, we discuss how HSP are linked to the ubiquitin-dependent proteasome system to generate peptides presentable by MHC molecules. PMID- 19551547 TI - The efficacy and safety of percutaneous microwave coagulation by a new microwave delivery system in large hepatocellular carcinomas: four case studies. AB - PURPOSE: To demonstrate the efficacy and safety of percutaneous microwave coagulation treatment (PMCT) by a new microwave delivery system (Forsea Microwave) in large hepatocellular carcinomas (HCC) (> or =5 cm). MATERIALS AND METHODS: Four patients with 4 HCC lesions measuring > or =6 cm in the greatest dimension underwent PMCT by means of the Forsea Microwave microwave delivery system. Final therapeutic efficacy was evaluated with dynamic computer tomography (CT) scans performed within one month after PMCT. During and after PMCT, patients' complaints and any abnormal physical signs were recorded for safety assay. CT or ultrasound scan (US) performed immediately after the treatment was used to detect acute complications related to the treatment. Repeated dynamic CT scans were performed every three to four months thereafter to detect local disease recurrence and/or other recurrences. RESULTS: Three of these patients achieved a complete ablation of the cancer nodules (two patients with two treatment sessions and one patient with three treatment sessions). One of these patients obtained a complete ablation of the cancer nodule with two treatment sessions except the lesion of portal vein tumour thrombus (PVTT). No obvious symptomatic complication was observed except abdominal pain during and after the treatment in two of these patients. All the patients remained asymptomatic and no recurrent tumour was observed during their follow-up (1-19 months). CONCLUSIONS: PMCT by the Forsea Microwave microwave delivery system could offer a satisfactory therapeutic effect and is applicable to the treatment of large HCC. PMID- 19551546 TI - Enhancement of hyperthermia-induced apoptosis by sanazole in human lymphoma U937 cells. AB - Sanazole has been tested clinically as a hypoxic cell radiosensitizer. The aim of the present study was to investigate whether sanazole enhances apoptosis induced by hyperthermia at 44 degrees C for 20 min in human lymphoma U937 cells. Sanazole alone induced continuous increase in the intracellular superoxide generation in a time-dependent manner and transient increase in the peroxide formation, which further were enhanced at 1 hour after HT treatment. Moreover, when the cells were treated first with 10 mM sanazole for 40 min, exposed to HT at 44 degrees C for 20 min and the cells were further treated with the drug at 37 degrees C for 6 h, a significant enhancement of HT-induced apoptosis was evidenced by DNA fragmentation, morphological changes and phosphatidylserine externalization. Studying the apoptotic pathways involved in this enhancement, we found that loss of the mitochondrial membrane potential, release of cytochrome c from mitochondria to cytosol, and activation of caspase-3 and caspase-8 was enhanced significantly in the U937 cells after the combined treatment. Moreover, this combination enhanced activation of Bid, and down regulation of Hsp70. In addition, an increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)), and externalization of Fas were observed immediately after sanazole and HT treatment. Our data indicate that sanazole can enhance the hyperthermia induced apoptosis through the Fas-caspase-8- and [Ca(2+)](i)-dependent apoptotic pathways. In addition, the down regulation of Hsp70 contributed to this enhancement. PMID- 19551548 TI - Climate change and occupational safety and health: establishing a preliminary framework. AB - The relationship between global climate change and occupational safety and health has not been extensively characterized. To begin such an effort, it may be useful to develop a framework for identifying how climate change could affect the workplace; workers; and occupational morbidity, mortality, and injury. This article develops such a framework based on a review of the published scientific literature from 1988-2008 that includes climatic effects, their interaction with occupational hazards, and their manifestation in the working population. Seven categories of climate-related hazards are identified: (1) increased ambient temperature, (2) air pollution, (3) ultraviolet exposure, (4) extreme weather, (5) vector-borne diseases and expanded habitats, (6) industrial transitions and emerging industries; and (7) changes in the built environment. This review indicates that while climate change may result in increasing the prevalence, distribution, and severity of known occupational hazards, there is no evidence of unique or previously unknown hazards. However, such a possibility should not be excluded, since there is potential for interactions of known hazards and new conditions leading to new hazards and risks. PMID- 19551549 TI - Heart rate, blood lactate concentration, and time-motion analysis of female basketball players during competition. AB - The aim of this study was to examine the physiological demands and movement patterns of female basketball players after changes in the rules of the game. Nine varsity players were studied during nine official games. Each game was videotaped to identify the frequencies of the main movements performed, heart rate was recorded continuously, and blood samples were collected to determine blood lactate concentration when the competition rules allowed. The main results showed that the players performed on average 652 +/- 128 movements per game, which corresponded to a change in activity every 2.82 s. Mean heart rate was 165 +/- 9 beats . min(-1) (89.1% of maximum heart rate) for total time and 170 +/- 8 beats . min(-1) (92.5% of maximum) for live time. Mean blood lactate concentration was 5.2 +/- 2.7 mmol . l(-1) (55.9% of maximum blood lactate concentration). In addition, heart rates were significantly higher in the first half than the second half of games. These results indicate: (1) a greater physiological load compared with previous studies on female players tested before the rules modification (Beam & Merrill, 1994; McArdle et al., 1971) and (2) lower movement frequencies compared with male players competing under modern rules (Ben Abdelkrim et al., 2007). These observations must be taken into account by coaches and conditioning specialists working with female players. PMID- 19551550 TI - A physiological, time-motion, and technical comparison of youth water polo and Acquagoal. AB - The aims of this study were to provide a profile of young water polo players and to compare technical and tactical aspects, movement patterns, and cardiac loads of youth water polo and Acquagoal codes. Ten young (age 12.3 years, s = 0.6) male water polo players underwent anthropometric (stature, body mass, body mass index, chest circumference, hand breadth, and length), strength, and [Vdot]O(2max) evaluations. Friendly youth water polo and Acquagoal matches were arranged to evaluate heart rates and swimming patterns (horizontal and vertical, with and without the ball) of players, and technical and tactical aspects of matches (number of actions, passes, player involved in an action, lost possessions, shots, goals, and the origin and types of shot). Independent of code, matches imposed a high cardiac load on players. Vertical swimming occurred more frequently (P < 0.05) in Acquagoal (71%) than youth water polo (45%). Technical and tactical measures also differed (P < 0.05), with players performing more passes and shots inside the penalty area and showing a higher goal-to-shot ratio during youth water polo than during Acquagoal. These results indicate that youth water polo better resembles the swimming patterns of adult water polo and facilitates the technical and tactical aspects of play better than Acquagoal. PMID- 19551551 TI - An in-depth assessment of expert sprint coaches' technical knowledge. AB - Among the myriad issues addressed by current research are the sources of knowledge of coaches in terms of their implicit and/or explicit natures. What appears to remain unaddressed, however, is the arguably prior step of coaches' content knowledge; that is, what coaches know as opposed to how they know it. The aim of this study was to partially address this shortcoming by identifying the characteristics that expert coaches associate with good sprint running technique, in addition to where such knowledge was gleaned. Seven expert male sprint coaches participated in the study, with data being gathered through in-depth, semi structured interviews. The data were inductively analysed using the ATLAS.ti 5.2 qualitative software. Four principal constructs emerged from the interview data. These included "posture", "hip position", "ground contact, and "arm action', with tacit, experiential sources, generated predominantly from conversations with respected peers, forming the basis of such knowledge. The findings are located within existing sprinting literature, before we summarize the main points made. PMID- 19551552 TI - Tactical use of the T area in squash by players of differing standard. AB - The importance of dominating the T in squash is recognized by coaches and players but there has been little formal investigation of this aspect of tactical play. Consequently, the aim of this research was to analyse player occupancy of a T area, to establish whether there are differences between winners and losers of games at different playing standards. An automated player-tracking system, with operator supervision and intervention, captured players' movements during matches at the World Team Championships (n = 11), the Slovenian National Championships (n = 11), and a local tournament (n = 15). Frequency of occupying the T area at the moment opponents played their shot best discriminated playing standard. Winners spent a greater proportion of total playing time in the T area than losers (P < 0.001), except during closely contested games. The results suggest that time in the T area indicates dominance of rallies. Future studies need to consider both between-group (playing standard) and within-game (individual player standard) differences, as both were shown to influence the time players spent in the T area. PMID- 19551554 TI - Is dysphonia due to allergic laryngitis being misdiagnosed as laryngopharyngeal reflux? AB - OBJECTIVES: Voice problems affect up to 30% of the population at some time during their life, and in particular it affects those who use their voice professionally. Patients who commonly present with dysphonia are often found to have symptoms of both allergy and reflux. Clinical examination of the larynx often makes it difficult to differentiate between the main causative factors of dysphonia, as the clinical findings on nasendoscopy between reflux and allergy are similar. Laryngopharyngeal reflux (LPR), an entity of gastro-oesophageal reflux, is a common diagnosis made in the voice out-patient clinic. Few studies have been able to successfully correlate management of LPR and outcome of dysphonia. Allergy or more specifically allergic rhinitis is said to affect approximately 24% of the population, but few studies have investigated its role in dysphonia. DESIGN: Prospective observational study. SETTINGS: University teaching hospital otolaryngology department. PARTICIPANTS: Fifteen new patients with primary voice disorder were investigated for possible LPR and allergy from April 2007 to October 2007. All patients were tested using the validated reflux symptom index (RSI), reflux finding score (RFS) to diagnose LPR, and both skin prick tests (SPT) and nasal nitric oxide (NO) levels to diagnose the presence of allergy. MAIN OUTCOME MEASURES: Prevalence of symptoms of reflux and allergy in patients presenting with primary voice disorders, and also whether dysphonia thought to be caused by LPR could actually be due to allergy. RESULTS: No positive correlation or statistical significance was found between reflux and allergy. From our sample 20% (three patients) were diagnosed with LPR and 67% (ten patients) with allergy. The three patients diagnosed with LPR also had concomitant allergy. CONCLUSIONS: In our patient cohort, three times as many patients demonstrated allergy compared with LPR. This has led us to question if some patients with allergic laryngitis are being misdiagnosed with LPR and thereby being over-treated with proton pump inhibitors (PPIs). PMID- 19551555 TI - Prevalence of depression in a general hospital in Izhevsk, Russia. AB - BACKGROUND: There are a lot of studies on depressive disorders in a general hospital done across the world, but no data from Russia on this subject was found in international psychiatric journals or MEDLINE. AIMS: to determine the prevalence of depressive disorders in medical inpatients in Izhevsk, the capital of the Udmurt Republic, a region in Russia, and to identify associated factors. METHOD: A sample of 323 adult medical inpatients was composed. The Russian version of the MINI 5.0.0 was used. RESULTS: The prevalence of lifetime and current depressive disorders was 30% and 20.7%, respectively. Depression was more common in women, widowed or divorced, retired or disabled, with low income and bad family relationships, and among respondents with a chronic somatic illness. Depression had a high comorbidity with organic mental and anxiety disorders. Only 40.3% of the individuals with depression had referred for psychiatric consultations, most of them being treated with fluvoxamine. CONCLUSIONS: Prevalence of depression was substantial but consistent with other studies. Taking into consideration associated factors, physicians can improve recognition and treatment of depression in medical inpatients. PMID- 19551556 TI - Antiangiogenic properties of prostate-specific antigen (PSA). AB - The prostate produces high levels of prostate-specific antigen (PSA, also known as kallikrein-related peptidase 3, KLK3), which is a potential target for tumor imaging and treatment. Although serum PSA levels are elevated in prostate cancer, PSA expression is lower in malignant than in normal prostatic epithelium and it is further reduced in poorly differentiated tumors. PSA has been shown to inhibit angiogenesis both in in vitro and in vivo models. In this review we focus on our recent studies concerning the mechanism of the antiangiogenic function of PSA. We have recently shown that the antiangiogenic activity of PSA is related to its enzymatic activity. Inactive PSA isoforms do not have antiangiogenic activity as studied by a human umbelical vein endothelial cell (HUVEC) tube formation model. Furthermore, inhibition of PSA, either by a monoclonal antibody or small molecule inhibitors abolishes the effect of PSA, while a peptide that stimulates the activity of PSA enhances the antiangiogenic effect. We have analyzed changes in gene expression associated with the PSA induced reduction of tube formation in the HUVEC model. Several small changes were observed and they were found to be opposite to those associated with tube formation. Taken together, these studies suggest that PSA exerts antiantiogenic activity related to its enzymatic activity. Thus it might be associated with the slow growth of prostate cancer. PMID- 19551557 TI - The role of glycodelin in cell differentiation and tumor growth. AB - Glycodelin is a lipocalin family glycoprotein expressed mainly in reproductive tissues. It is involved in cell recognition, and its relationship with epithelial differentiation is well established. Glycodelin actually appears to drive epithelial differentiation. The evidence comes from studies employing endometrial and breast cancer cell lines. First, transfection of glycodelin cDNA into glycodelin-negative carcinoma cells results in reduced expression of oncogenes, increased expression of tumor suppressor genes, increased cell differentiation, and reduced carcinoma cell growth. Second, histone deacetylase inhibitors (HDACIs) induce glycodelin synthesis in endometrial cancer cells concomitantly with cell differentiation. This effect is blocked by specific down-regulation of glycodelin by RNA interference, suggesting that the effects of HDACIs are mediated by glycodelin. We recently found that glycodelin not only reduces carcinoma cell growth in vitro, but glycodelin cDNA transfection to MCF-7 breast carcinoma cells also reduces growth of these cells in vivo, demonstrated by xenograft tumor growth in mouse mammary fat pads. These results strongly suggest that glycodelin acts as a tumor suppressor in breast cancer. The findings are compatible with the observations that certain types of glycodelin-expressing ovarian and breast cancers have a more favorable prognosis compared to glycodelin non-expressing tumors. This research has therefore introduced a novel mechanism to control cancer cell growth. In this communication we review the differentiation-related effects of glycodelin. PMID- 19551559 TI - Orbital approach for retrieval of transected extraocular muscles. AB - Transection of an extraocular muscle can occur from orbital and facial trauma, or as a complication of surgery. The injury can occur either near the muscle insertion or in the muscle belly. Identification of the proximal end of the muscle in the orbit may be difficult, especially if the transection occurs a farther distance from the insertion, and, in these cases, the muscle is often considered lost. We present two patients who suffered from traumatic transections of an extraocular muscle more than 10 mm from the insertion. Both patients underwent transconjunctival orbitotomy to retrieve and secure the severed extraocular muscle. Both patients achieved good primary gaze alignment postoperatively. Preoperative imaging should be considered in cases of traumatic extraocular muscle transection. PMID- 19551560 TI - Muscle belly union for treatment of myopic strabismus fixus. AB - PURPOSE: To evaluate the surgical results of the medial rectus (MR) muscle recession associated with longitudinal splitting of the lateral rectus (LR) and superior rectus (SR) muscles and surgical union of their corresponding midpoints in highly myopic patients with severe esotropia and hypotropia and restricted abduction and elevation. METHODS: Six cases of severe high myopic strabismus fixus were considered with more than 90Delta esotropia and 25Delta to 30Delta hypotropia. The nasally deviated SR muscle and inferiorly shifted LR muscle were confirmed by MRI or CT scan. Supratemporal herniation of the globe from the muscle cone was also found by imaging. The LR and SR muscles were split in half from the insertion to past the equator. The lateral half of the SR muscle was united to the superior half of the LR muscle. Medial rectus muscle was recessed also. RESULTS: Motility gradually improved and by 8 weeks in four cases there was 5Delta to 15Delta residual esotropia and no hypotropia and mild limitation in abduction and elevation. In two cases due to marked residual esotropia, we performed a second operation, and 2 months later residual esotropia was mild. CONCLUSION: In highly myopic patients, if the deviant paths of the LR and SR muscles are demonstrated by MRI or CT scan, the surgical method described is effective and recommended. PMID- 19551561 TI - Asperger syndrome associated with idiopathic infantile nystagmus--a report of 2 cases. AB - Asperger syndrome is a severe and chronic developmental disorder. It is closely associated with autism and is grouped under autism spectrum disorder (ASD). Various eye movement abnormalities in AS have been reported in literature such as increased errors and latencies on the antisaccadic task implicating dysfunction of the prefrontal cortex, impairment of the pursuit especially for targets presented in the right visual hemisphere, suggesting disturbance in the left extrastraite cortex. There are no reports in the literature of association between idiopathic infantile nystagmus (IIN) and AS. We report 2 cases of Asperger syndrome associated with idiopathic infantile nystagmus. PMID- 19551562 TI - Case report and literature review of inferior rectus muscle aplasia in 16 Japanese patients. AB - PURPOSE: To describe clinical features of inferior rectus muscle aplasia in Japanese patients and to gain insight into its pathogenesis. METHODS: Case presentation and literature review of 16 Japanese patients with inferior rectus muscle aplasia. RESULTS: The age at presentation of 16 patients (9 females, 4 males, and 3 with unknown gender) varied from 7 months to 73 years (mean, 21.8 years). The inferior rectus muscle was absent on the right side in 10 patients, on the left side in 4 patients, and on both sides in 2 patients. Structural anomalies of the eye globe, such as microphthalmos, microcornea, and coloboma of varying degrees, were present on the ipsilateral side of the inferior rectus muscle aplasia in 4 patients and on the contralateral side in 1 patient. As surgical findings, anomalous inferiorly located insertion of the medial rectus muscle was discovered in 5 patients: 1 patient in association with microcornea and iris coloboma. Frequent surgical procedures were recession or tenotomy of the superior rectus muscle, combined with downward transposition of the entire or partial tendons of the medial rectus and lateral rectus muscle to the medial and lateral end, respectively, of the putative inferior rectus muscle insertion. CONCLUSIONS: The association of coloboma with inferior rectus muscle aplasia suggests that abnormal optic fissure closure during embryogenesis might underlie the muscle aplasia. PMID- 19551563 TI - PHACE syndrome associated with congenital oculomotor nerve palsy. AB - PHACE syndrome is a multisystem disorder presenting with facial hemangiomas, arterial anomalies, cardiac anomalies, posterior fossa malformations and eye abnormalities. The eye abnormalities include microphthalmos, cataracts, optic atrophy and iris hypoplasia. Amongst the neurological anomalies, posterior fossa malformations are common. Fourth nerve palsy has been reported with PHACE syndrome. We report a child presenting with a triad of congenital third nerve palsy, cerebellar hypoplasia and facial capillary hemangioma. PMID- 19551564 TI - An audit of the Royal College of Ophthalmologists strabismic amblyopia treatment protocol: a departmental review. AB - INTRODUCTION: Children presenting with strabismus and mixed (anisometropic/strabismic) amblyopia are managed by a local protocol as per guidelines from the Royal College of Ophthalmologists. Decisions regarding intervention for occlusion are currently delayed until a 22 week review allowing for refractive adaptation, with intermediate reviews at 6 and 14 weeks. PURPOSE: The purpose of this audit was to determine adherence to the protocol and the benefit of the 14 week review. MATERIALS AND METHODS: We performed a prospective data collection of all children attending the orthoptic department with strabismus without pathology, both with and without unequal vision, from October 2007 to July 2008 managed using the protocol. RESULTS: 26 patients were eligible. Mean age at presentation was 3.3 years (1.4 to 6.5). Cycloplegic spherical equivalent (SE) mean was 2.6 dioptres (-2.25 to +7.25). Five patients failed to comply with the protocol; one patient was listed surgery, four patients commenced premature occlusion. At presentation 8 patients had equal vision (defined as < 0.1 logMAR difference) between the two eyes or would only perform BEO vision, by week 14, over 60% were found to have a difference in vision between the eyes, despite refraction correction (mean 0.4 logMAR, range 0 to 1.4 logMAR). In a subgroup of patients (n = 8 at 6 weeks) with a small discrepancy of vision between the eyes (0.1 to 0.5 logMAR) there was no progressive worsening of vision during the period of observation and 50% of patients improved spontaneously, although two patients were lost to follow up. In the subgroup (n = 10 at 6 weeks) with a large discrepancy between the eyes (> 0.5 logMAR), all patients ultimately required occlusion, and there was minimal improvement in only three patients. CONCLUSION: Recorded vision at 6 weeks is more informative for making treatment decisions than that recorded at presentation. The 14 week review confers no benefit to those with reliable and improving visual acuity. Those patients presenting with a large discrepancy in visual acuity do not improve after 14 weeks and we feel that these patients could be occluded at 6 weeks. We propose a new algorithm for the treatment of this patient group. PMID- 19551565 TI - Drift of visually induced optostatic torsion. AB - PURPOSE: Compensatory ocular torsion has been found in response to static tilted visual stimuli. The purpose was to investigate if visually induced ocular torsion is maintained during continuous stimulation and to reinvestigate the effect of stimuli tilt angle. METHODS: Eye movements were recorded in 19 healthy individuals using a 3-D video oculography system. One visual stimulus was used in the tests, a photographic image of a city scene with spatial clues relevant for body posture. The scenes were tilted counterclockwise around a central pivot point from 0 to 15 degrees (test A), from 0 to 30 degrees (test B), and from 0 to 45 degrees (test C). Each tilted stimulus position was held static for 4 min, 40 sec. RESULTS: A compensatory torsional response was found in all subjects and test conditions. No significant difference was found between the different stimuli tilt angles. The torsional response was not maintained but drifted back towards the reference position. CONCLUSIONS: Visually induced ocular torsion in response to a static tilted scene is not maintained, but drifts back towards the initial reference position. Higher cognitive functions such as imagination and attention certainly have an influence on the outcome. PMID- 19551566 TI - Experiences with the elimination of amblyopia and the restoration of binocular vision in squinters, part I. 1927. PMID- 19551569 TI - [Carpal tunnel syndrome as a possible occupational disease. Neurologic assessment]. PMID- 19551570 TI - [Carpal tunnel syndrome as possible occupational disease. Occupational medicine assessment]. PMID- 19551571 TI - [In memoriam--surgeon of the century]. PMID- 19551578 TI - [The German trauma network--current status]. PMID- 19551579 TI - [Fatal risk thrombosis? A prospective study for the incidence of deep vein thrombosis in pelvic fractures]. AB - AIM: The aim of the study was to determine the incidence of deep vein thrombosis (DVT) after pelvic trauma and surgical stabilisation of pelvic and acetabular fractures under medicamentous prophylaxis. PATIENTS AND METHOD: Within 20 months we prospectively included 50 patients. 25 had an isolated pelvic trauma, 25 patients had multiple injuries. 21 of them were polytrauma patients (average ISS: 31.4; min. 26 pts., max. 50 pts.), four patients had additional highly unstable spine fractures or fractures of the lower extremities. Low molecular-weight heparin (Enoxaparin 40 g/d) was administered on average within 24 hours of injury in 44 cases, one patient received low-dose heparin (Liquemin 15,000 to 22,500 I. E./d), five patients received both. 31 patients were treated operatively and 19 conservatively. Colour-flow duplex ultrasonography was performed within 72 hours of injury and stabilisation of the pelvic and acetabular fracture, or weekly. By means of ultrasound, 97 to 100 % of the deep and superficial leg veins could be examinated safely, as well as 88 to 89 % of the external iliac veins and 64 to 66 % of the common iliac veins. Only in 36 to 40 % of the patients the internal iliac veins were visible by ultrasound. RESULTS: Proximal DVTs were detected postoperatively in two patients (4 %), one patient (2 %) died after a fatal P. E. before the scheduled duplex scan. CONCLUSION: Early medicamentous prophylaxis can prevent deep vein thrombosis after pelvic trauma. Delayed applications due to pelvic operations are risk factors. In such cases duplex scanning should be performed routinely and postoperative medicamentous prophylaxis should be increased. PMID- 19551580 TI - [The articular fracture of the lower limb]. AB - AIMS: The combination of a distal tibia fracture with an additional upper ankle joint injury is a challenge. Apart from various implants the intraoperative problem is the handling of these two injuries with appropriate reduction and retention. The existing and further developing soft tissue damage has to be taken into consideration. The aim of this study is to evaluate the surgical management of this type of fracture. PATIENTS AND METHODS: Between 2000 and 2007 we treated 22 patients with tibia fractures and coexisting upper ankle fractures. All patients could be traced for follow-up examinations. We deduced the impact energy of the trauma and the soft tissue damage. The tibial and ankle fracture sites were categorised. The time elapsed until full weight bearing was measured. The ankle joint function was inspected. RESULTS: A high energy trauma existed in 10 and a low energy trauma in 12 patients. In the high energy group we found in 5 cases A3, in 2 cases B1 and 1 B2 and 1 B3 and 1 C2 fracture of the tibia. In 5 cases a bimalleolar ankle injury existed, once a medial ankle fracture und in 3 cases an isolated distal fibula fracture. One patient had an open ankle joint luxation. In some cases an additional fibula shaft fracture was seen. "Fracture communication" between the tibia fracture and the ankle injury was not seen at all. An intramedullary nail stabilisation was used in 9 cases, while in 1 patient due to the soft tissue damage an external fixator was applied. In the low-energy group there were 4 B1, 4 B2 and 4 C1 fractures of the tibia. We found 6 injuries of the posterior plafond and 6 cases with a central fracture line of the pilon. A fracture communication between ankle and tibial shaft was detected in all cases. In 7 patients we used a minimally invasive locking plate and in 5 cases a nailing technique for stabilisation of the tibia fracture. Overall, we saw 1 distraction fault, 1 valgus misfitting of the fracture, 1 pin infection and 2 soft tissue necrosis as postoperative complications. The time interval until osseous union was 3.5 months. Monoarticular fractures of the upper ankle joint had better results according to the Weber score. CONCLUSION: One can divide the distal articular tibial shaft fracture into two groups. In the high energy entity the ankle joint injury happens first, and afterwards the tibial shaft fracture occurs. Therefore, both fracture sites are usually not communicating, which means they are in fact two types of fracture. On the other hand, in the low energy group, both fractures are communicating. Here, the tibial shaft fracture is equal to the inner ankle fracture in a classic bimalleolar fracture. Therefore we have only one fracture site. PMID- 19551581 TI - [Unstable pertrochanteric femur fractures. Failure rate, lag screw sliding and outcome with extra- and intramedullary devices (PCCP, DHS and PFN)]. AB - AIM OF THE STUDY: The dynamic hip screw (DHS) often shows a high incidence of therapeutic failure and an impared outcome in the treatment of the unstable pertrochanteric femur fracture (31A2). Therefore often an intramedullary device is recommended. In a retrospective clinical study we examined whether the percutaneous compression plate (PCCP, Gotfried) provides advantages following unstable fractures in comparison to DHS and PFN. METHODS: From January 2002 to April 2007 135 patients with unstable pertrochanteric femur fractures underwent internal fixation with the PCCP (n = 46, age 78.3, ASA 2.8), DHS (n = 36, age 75.9, ASA 3.0) or PFN (n = 53, age 77.2, ASA 2.8). Radiological and clinical re examination of the patients (33 PCCP, 24 DHS, 34 PFN) was performed 17 months later. RESULTS AND DISCUSSION: The PCCP was implanted in less time than the DHS and PFN (59 vs. 80 vs. 79 min, p = 0.004). Radiographic screening time was low (PCCP 143 vs. DHS 146 vs. PFN 280 s, p = 0.001). Re-operations for wound infections and haematomas occurred in 2 % after PCCP, 14 % after DHS and 4 % after PFN (p = 0.058). There was a low re-operation rate for fracture fixation complications in PCCP (9 %), in contrast to DHS (25 %) and PFN (13 %, p = 0.109). Cut-out was seen more in DHS (19 %, PCCP 2 %, PFN 4 %, p = 0.005). Lag screw sliding was high with DHS (PCCP 4 mm vs. DHS 9 mm vs. PFN 6 mm, p = 0.032). There was no correlation between lag screw sliding and outcome. PCCP, DHS and PFN had the same functional results in Merle d'Aubigne and Harris hip scores. CONCLUSIONS: Using the minimally invasive PCCP technique in unstable pertrochanteric femur fractures provides a promising therapy option especially with regard to surgical time, radiographic screening time and failure rate. Lag screw sliding was low. There was no advantage of the intramedullary device PFN. PMID- 19551582 TI - [Correction of supramalleolar deformities with the Taylor spatial frame]. AB - BACKGROUND: Post-traumatic supramalleolar deformities and malunions of ankle fusion require mostly a multiplanar correction. In cases of severe soft tissue damage, external fixation and gradual correction is a definite treatment alternative. METHOD: Between 2003 and 2007 a correction of supramalleolar deformities was performed in 9 patients with the Taylor spatial frame external fixator. The mean age was 30 years (min. 12, max. 68). There were 6 patients with deformities after malunion of supramalleolar fractures and 3 patients with malunion after ankle fusion. The mean angular deformity was 30 degrees and 5 patients had a rotational malposition of 13.6 degrees (min. 5 degrees, max. 25 degrees). 5 patients needed also lengthening (min. 10 mm, max. 40 mm) of the post traumatic deformed tibia. The mean time of the follow-up examination was 23 months (min. 12, max. 41). RESULTS: Anatomic correction could be achieved in all patients. The average correction time was 36 days (min. 10, max. 82) with an average time period of 163 days (min. 130, max. 218) until the fixator was removed. The healing index of the leg lengthening cases was 77 d/cm. There were 2 pin-tract infections, 1 prolongated callus formation and 1 insufficient callus formation. CONCLUSION: In cases of post-traumatic supramalleolar deformities with poor soft tissue and bone quality the Taylor spatial frame is a useful tool. Angular, axial, translational and rotatory deformities are corrected simultaneously without complex and time-consuming fixator reconstructions. PMID- 19551583 TI - [Operative and follow-up treatment of rotator cuff tears--the current situation]. AB - AIM: The aim of this study was to evaluate the current situation of rotator cuff repair and follow-up treatment in German hospitals. METHODS: An evaluated survey, enquiring about the year 2006, was sent to all 777 German orthopaedic and/or trauma surgery departments. The hospitals were chosen using the official index of hospitals from the German Federal Statistical Office. RESULTS: 44 % of the surveys were sent back, whereby 40 % were of use. Within the 309 departments, 26 % of the total number of 59,957 shoulder operations were rotator cuff repairs. Mini-open was the operation method in 49 %, open in 29 % and arthroscopic in 22 % of cases. Regarding the operational methods, there were differences between the departments and level of care. The arthroscopic technique was used sometimes in 48 % of the departments, whereas 52 % never used it. Specialised shoulder departments employed the total arthroscopic procedure more often. A set follow-up treatment occurred in 79 % of departments. CONCLUSION: Rotator cuff repair is mainly carried out using the mini-open method, which is nowadays regarded as gold standard. PMID- 19551584 TI - [The percutaneous bone biopsy: in vitro study for comparison of bone biopsy needles]. AB - PURPOSE: The aim of this study was to assess the differences in ease of use and quality of samples of several bone biopsy needles in an animal test. MATERIALS AND METHODS: An evaluation of eight bone biopsy needles of different gauges was undertaken. With each needle, 5 biopsies of an animal bone (lumbal vertebral body of calf, pig and lamb) were performed and compared to each other. The subjective assessment of force to obtain a sample, ease of needle use and ease of sample removal were graded on a 5-point scale. Each biopsy specimen was measured before and after fixation and the gross state was evaluated. For evaluation of histopathological quality, width and degree of fragmentation were also evaluated on a scale. RESULTS: The Somatex, Bone Marrow and Safe Cut 8 G and the Cardinal Health, Jamshidi 8 G needles were rated as being the easiest ones to use, while the Bloodline, Easy Trap 8 G and the RADI, Bonopty 15 G biopsy needles were rated as being the most difficult ones. Histological specimen quality was highest for the Somatex 8 G needles, the Cardinal Health, Jamshidi 8 G and the Bloodline, Easy Trap 8 G needles. The Inter.V, SnareLok 8 G and the RADI, Bonopty 15 G needles had the lowest yield. Furthermore, differences in length before and after fixation were recorded. The average decrease of core length after fixation was 18 %. CONCLUSION: The bone biopsy needles tested here vary significantly in performance and quality of the histopathological specimen. Detailed knowledge of the strengths and weaknesses of different needles could facilitate the decision for the selection of an appropriate instrument. PMID- 19551585 TI - [Transfixation in slipped capital femoral epiphysis: long-term evidence for femoro-acetabular impingement]. AB - AIM: It was the aim of this study to reveal whether and to what extent femoro acetabular impingement (FAI) was present 10 to 20 years after transfixation of the gliding epiphysis in patients treated for unilateral slipped capital femoral epiphysis (SCFE). METHOD: Clinical and radiological evaluations were undertaken of 20 patients who were treated with a bilateral transfixation of the epiphysis with screws or Kirschner wires because of mild unilateral SCFE. The Harris hip score, the activity level according to Tegner and Lysholm and the range of motion of both hip joints were noted. An impingement provocation test was performed on both sides. The femoral head-neck junction was qualitatively and quantitatively assessed in anteroposterior radiographs of the pelvis and "cross table" lateral radiographs of both hip joints. Quantitative assessment was achieved by measuring the alpha angle according to Notzli et al. RESULTS: All except one patient showed excellent results in the Harris hip score. The activity levels of the majority of patients, however, were mediocre. There was no difference for flexion, whereas internal rotation and abduction significantly differed between affected and non affected hips (p = 0.135, p = 0.002 and p = 0.007, respectively). The impingement provocation test was found positive in eight affected and five non-affected hips (p = 0.004). The alpha angle was significantly different between affected and non affected hips in the anteroposterior radiographs of the pelvis (86 vs. 61 degrees; p < 0.001), but not so, however, in the "cross table" lateral radiographs (52 vs. 46 degrees; p = 0.145). CONCLUSION: Clinical evaluation of 20 patients at a mean of 14.9 years after transfixation of the epiphysis revealed some evidence for the presence of FAI in affected hip joints. Radiological evaluation suggests that its localisation is more lateral and less anterior. PMID- 19551587 TI - [Antibiotic prophylaxis for patients with joint prostheses undergoing dental treatment--a topic for discussion]. AB - AIM: In accordance with international guidelines, the German Society for Cardiology and the Paul Ehrlich Society for Chemotherapy recently adapted their recommendations for antibiotic prophylaxis of infectious endocarditis. The new version reflected the statistically lower risk for such infections as compared to former considerations and reduced the group of patients who would benefit from the prophylaxis. That paper as well as an increasing number of statements of orthopaedic/traumatologic or dental societies stimulated our contribution on the prevention of prosthesis infections after dental care. With this article we intend to stimulate a position paper of the German Society for Orthopaedics and Traumatology. METHOD: For our study we screened the international literature on the association between bacteremia and dental treatment, bacteremia and prosthesis infections as well as on the availability and risks of antibiotic prophylaxis for prosthesis infections. In addition, we included data on the responsible microorganisms and the importance of biofilms both in the oral cavity and on the infected prosthesis. RESULTS: Generally, the risk of prosthesis infections after bacteremia is lower than that of endocarditis. Also, the range of involved microorganisms only partially overlaps in both diseases. Of note, bacteremia regularly occurs due to normal dental hygiene measures or even after chewing. Because of this high background risk and because of the extended latency period between dental care and symptomatic prosthesis infections, the causality of professional dental measures for prosthesis infections has never conclusively been demonstrated, e.g., by employing molecular methods. However, the association remains plausible and the consequences for such patients are severe. CONCLUSION: We suggest an oral prophylaxis with an aminopenicillin plus beta-lactamase inhibitor or clindamycin shortly before and 4 hours after dental care depending on the tissue invasiveness of the dental measures and the personal risk profile of the patient (prosthesis recently implanted, history of prosthesis infection, natural or iatrogenic conditions severely affecting the immune status). PMID- 19551586 TI - [Hip centralising forces of the iliotibial tract with various femoral neck angles]. AB - AIM: Concerning the biomechanical properties of the iliotibial tract different opinions can be found in the literature. Due to this fact it was the aim of this study to take measurements about the hip centralising forces of the iliotibial tract in a neutral position as well as in abduction, adduction and flexion of the hip joint at different femoral neck angles by using a custom-made hip prosthesis. METHOD: By using a custom-made measuring endoprothesis with the capability for adjusting different femoral neck angles and lengths and, furthermore, by measuring the intra- and subligamentous forces at the height of the greater trochanter, we measured the forces for validating the influence of the iliotibial tract for hip joint centralisation. RESULTS: By increasing the CCD angle (coxa valga) a higher load on the hip joint results. By decreasing the CCD angle (coxa vara) a lowering of the coxalfemoral load results. Flexion of the knee joint leads to a decrease of the iliotibial tract tension. By extension of the knee joint and resulting tightening of the iliotibial tract, a triplication of the forces at the femoral neck was measured. Flexion of the hip joint exerted a ventralising of the iliotibial tract with an initial decrease of the centralising hip forces. Subligamentous measurement of the iliotibial tract showed increasing forces upon adduction as well as decreasing forces upon abduction of the hip joint. In the investigations about the forces with various femoral neck lengths, we saw a considerable increase by lengthening the femoral neck and resulting higher forces in the acetabulum. CONCLUSION: The clinical relevance of these results concerns the predictability of the in- or decreasing tension band wiring effect of the iliotibial tract in correlation to the CCD angle and the direction of motion of the hip joint. The measurements give the clinical users a benchmark for the expected subligamentous forces of the iliotibial tract and the resulting hip centralising forces. The influence of the lengthening of the femoral neck for the hip centralising forces clarifies the importance of the iliotibial tract when planning displacement osteotomies or hip joint replacement. PMID- 19551588 TI - [Triple arthrodesis in treatment of degenerative hindfoot deformities: clinical, radiological and pedobarographic results]. AB - INTRODUCTION: Triple arthrodesis is performed to reconstruct a painless plantigrad foot in cases of fixed and painful hindfoot deformity. Mid-term results of our patients after triple arthrodesis concerning clinical and functional as well as radiological outcome were assessed in this examination. PATIENTS AND METHOD: In the retrospective study, 24 feet in 20 patients with triple arthrodesis after degenerative hindfoot malposition were examined. Sampling was done consecutively. The average age of the patients was 62.8 years (40-79), mean follow-up-period was 21.5 months. The patients were examined using the AOFAS score, assessment of subjective satisfaction and X-rays. In addition, the position of the foot was analysed using dynamic pedobarography and gait analysis. RESULTS: In the examined population, the mean AOFAS score was 74 (+/- 12.41) out of 94 points, the improvement averaged 51 (+/- 14.72) points. 75 % of the patients had no or only slight pain, 8 % reported limitations in everyday life. Mobility of the ankle was more than 30 degrees for 9 patients (38 %) and between 15 and 30 degrees for 13 patients (54 %). 23 patients out of 24 were satisfied with the result of the surgery. In most cases, dynamic pedobarography showed a rather physiological pattern of plantar pressure distribution with increased pressure remaining in the midfoot area. Valgus position of the hindfoot was 2.8 degrees (+/- 2.9 degrees), gait analysis showed very good results in 11 cases and good results in 13 cases. Among the examined patients, two cases of wound healing disorders appeared, and one non-union of the talonavicular joint which had to be reoperated. In two cases, clinically relevant degenerative changes of the ankle were diagnosed. CONCLUSION: Triple arthrodesis using internal fixation followed by sufficient immobilisation is a good and reliable technique for the correction of fixed hindfoot deformities. A very high level of patient satisfaction and a good clinical outcome can be achieved. Plantar pressure distribution can be reconstructed to a satisfactory extent. Due to the relatively high rate of degenerative arthritis in adjacent joints, the decision upon performing a triple arthrodesis should be considered carefully. Clinical and radiological integrity of these joints are required. PMID- 19551589 TI - [Painful enlargement of the 2nd toe due to an osteoid osteoma in the distal phalanx]. AB - The rare case of an osteoid osteoma in the distal phalanx of the 2nd toe resulting in painful enlargement and hypertrophy of the entire toe in a 12-year old girl is discussed. The tumour was excised and the oversize of the toe was corrected by exarticulation of the distal phalanx. 18 months postoperatively the patient demonstrates normal function of her forefoot without complaints or signs of inflammation. PMID- 19551590 TI - [Ochronosis--a rare cause of secondary gonarthrosis]. AB - Ochronosis is a manifestation of the rare disease alkaptonuria. The most common presentations include pain in the lumbar spine region starting during the 3rd decade, spreading over the large joints. There exists no curative therapy for the disease at the moment. In the long-term the patients will be dependent on total joint arthroplasty. PMID- 19551591 TI - [The interesting case: schwannoma of the nervus pudendus after triple pelvic osteotomy according to toennis]. AB - Schwannomas (neurilemmomas) are benign neural sheath tumours which commonly arise from cranial nerves and cutaneous nerves of the head and neck. The most common site is the acoustic neuroma of the 8th cranial nerve. Pelvic schwannomas are rare and often present with non-specific symptoms leading to misdiagnosis and prolonged morbidity. Most cases of pelvic schwannoma have been reported in the gynaecological and urological literature due to their presentation as a pelvic mass or from urinary tract compression. We present a schwannoma of the nervus pudendus with clinical, radiological, MRI scan and intraoperative findings together with a description of the technique of surgical resection. PMID- 19551593 TI - [Humerus shaft fractures]. AB - Fractures of the humerus shaft are a common injury, mainly in younger patients and often coming with other injuries. The strategies of treatment are the conservative therapy, which was favoured till the late 1970s, and the operative therapy, including intramedullary nailing, plate osteosynthesis and external fixation. The initial procedure should be chosen according to the primary soft tissue damage and the existence of radial nerve palsy. The aim of treatment must be a usable extremity with anatomical axis, length and rotation. PMID- 19551595 TI - Psychiatry for neurologists. Preface. PMID- 19551592 TI - [The dorsal approach to the radiocarpal joint: an anatomic variant and its clinical implications]. AB - The extensor carpi radialis longus tendon and the extensor carpi radialis brevis tendon are important landmarks for the dorsoradial approach to the wrist. This case report presents an anatomic variant: both tendons are divided into two different reins. Knowledge about this anatomic variant is of importance in order to avoid misinterpretations. PMID- 19551596 TI - The neuropsychiatric mental status examination. AB - Neuropsychiatric disorders can cause a wide range of effects. Many aspects of behavior, cognition, and personality can be impacted. This article illustrates sensitive interview techniques for obtaining structured pertinent information to capture the effects of neuropsychiatric disorders. It then goes on to explain the components of the cognitive evaluation, with an emphasis on the hierarchical nature of cognition. It includes practical comments about evaluating cognitive dysfunctions, along with recommendations regarding the use of practical batteries. The mental status examination is then presented in detail, including nuances in assessing activity, mood, and perceptual disturbances. The manuscript content provides a template for the overall evaluation of neuropsychiatric patients. PMID- 19551597 TI - Psychotherapy for neurologists. AB - Psychotherapy has traditionally been regarded as the purview of psychiatry rather than neurology. Yet, the doctor-patient relationship is fundamental to both specialties, and the principles that derive from psychotherapy theory and practice apply to that relationship regardless of the specialty. It is common knowledge that a large proportion of patients seen in the context of the practice of medicine have some kind of emotional disturbance. Moreover, patients with organic disease may also have significant emotional difficulties that complicate both the primary illness and its treatment. This experience inevitably has drawn attention to the need for the nonpsychiatric physician to have an understanding and proficiency in psychiatric diagnosis and psychotherapeutic principles. In this article, we consider basic psychotherapeutic principles that are useful in the everyday practice of neurologists and other nonpsychiatric physicians. These skills are important not only for practical reasons, but also because responsiveness to their emotional distress is essential to maintain empathy and caring as cornerstones of the art of medicine. With the use of clinical examples to illustrate these principles, we hope that readers can apply them to their own clinical experiences. PMID- 19551598 TI - Psychopharmacology for neurologists. AB - Psychiatric disorders are common in neurological patients, and psychopharmacological agents are frequently used to treat agitation and other problems in neurological practice. Antidepressants are effective for depression caused by neurological illness, but they can interact with other medications. Antianxiety drugs can aggravate cognitive disorders and should be used cautiously in this context. Antipsychotic drugs can be useful for acute agitation, but they are being found to be neither effective nor safe for chronic nonpsychotic agitation. Other agents such as beta blockers and serotonergic agents are being found to be more useful. Uses, doses, adverse effects, and interactions of psychiatric medications in neurology are summarized in this article, and suggestions are provided for the practical application of these treatments. PMID- 19551599 TI - Depression in neurological practice: diagnosis, treatment, implications. AB - Ambulatory prevalence rates for significant depressive syndromes in general neurology clinics are quite high, in the range of 15 to 20% of clinic attendees. These depressive syndromes are a source of considerable morbidity and even mortality for the patients who suffer from them. Depression is a treatable syndrome, but there are not enough psychiatrists to administer all the treatments. Inevitably, many neurologists will become involved with some antidepressant therapies. In this article, I review a series of steps that can be used by neurologists to diagnose and treat the depressive disorders that occur in their practices. The Goldman algorithm for the treatment of depression is also presented as a therapeutic tool for practicing neurologists. PMID- 19551600 TI - Somatoform disorders. AB - The fields of neurology and psychiatry were originally practiced as a unitary model. As the fields dichotomized patients into "organic" and "functional," patients with medically unexplained symptoms fell between the borderlands. Despite their significant prevalence, somatoform disorders (SDs) are grossly under-recognized and lack effective treatments. This article reviews diagnostic criteria, differential diagnoses, examination findings, and treatments for these disorders. A summary of advances in research in functional neuroimaging and treatment trials is also given. PMID- 19551601 TI - Neuropsychiatric sequelae of traumatic brain injury. AB - The prevalence of traumatic brain injury (TBI) is increasing, particularly in the population of veterans. Many times, the motor and sensory consequences of TBI are addressed, but the post-TBI neuropsychiatric sequelae, which may be as, or even more devastating than the motor and sensory deficits, are left unattended. Cognitive, mood, anxiety, thought, impulse, and substance disorders, and a variety of personality disorders can be seen following TBI. The neuropsychiatric sequelae of TBI not only interfere with day-to-day function, but can severely impede rehabilitation efforts. To date, there have been few large-scale studies looking at the effectiveness of the various treatment modalities, including psychotherapeutic and pharmacological interventions. PMID- 19551602 TI - Psychiatric neurosurgery 2009: review and perspective. AB - In the treatment of psychiatric disorders, modern lesion procedures and nonablative deep brain stimulation (DBS) offer a degree of hope for patients who remain severely ill and impaired despite pharmacological and behavioral treatments. The available data support the therapeutic promise of these procedures, primarily for those suffering with obsessive-compulsive disorder (OCD) or major depressive disorder (MDD). However, the use of neurosurgical treatments for psychiatric disorders must be approached with both caution and a commitment to long-term care. The data also show that psychiatric neurosurgical procedures can be implemented most successfully by dedicated interdisciplinary teams in the context of a multimodal treatment plan. Treatment using these procedures is further complicated by issues involving the criteria for patient selection, the long-term management of patients receiving psychiatric neurosurgery, and the different patterns of potential clinical benefits and burdens presented by DBS and contemporary lesion procedures. PMID- 19551603 TI - Personality disorders: understanding and managing the difficult patient in neurology practice. AB - This article provides a background for understanding and managing maladaptive personality traits and personality disorders in neurology practice. These characteristics are commonplace in neurology patients and may cause diagnostic confusion, increased functional impairment, and complications in the doctor patient relationship. Maladaptive personality traits and personality disorders may precede neurological illness, may contribute to circumstances that lead to neurological injury, and may be caused by neurological illness, or some combination of these factors. Maladaptive personality traits associated with key neurological illnesses are reviewed, as are the major personality disorders, maladaptive defense mechanisms, countertransference reactions, and how these combine to contribute to difficulty in patient management. Finally, basic clinical management strategies are suggested. PMID- 19551604 TI - [Care of the dying in the hospital: initial experience with the Liverpool Care Pathway (LCP) in Germany]. AB - BACKGROUND AND OBJECTIVE: In the last few years public interest in the care of severely ill and dying patients has been growing. The aim of palliative medicine is to improve the care of the dying. However, this is still not achieved in many general hospitals. The Liverpool Care Pathway (LCP) for the care of the dying intends to change this situation. The aim of this study was to explore the views of professionals using the LCP as a framework for ensuring good care of the dying. METHODS: A qualitative study was conducted with an interdisciplinary focus group of ten professionals (nurses, physicians, spiritual adviser, social worker, physiotherapist and art therapist) to explore their views and experience after implementation of the LCP in a palliative care unit (PCU). The recorded discussion between them was transcribed verbatim and analysed using content analysis by three independent reviewers. RESULTS: Seven nurses and three physicians with an average work experience of 16 years each took part in the focus group. Based on the experience of 24 patients, the LCP was evaluated as very positive by all participants. In particular, three aspects were emphasized as having high relevance for a good quality of care: improvement of self confidence, better control of symptoms, and enhancement of the communication between professionals and with patients and their relatives. However, some weaknesses were also mentioned, e.g. inadequate effort of documenting the beginning of implementing the scheme. CONCLUSION: The LCP was well received by professionals after the initial implementation of the LCP in a German PCU. The LCP was judged as an appropriate and helpful framework in the care of the dying. PMID- 19551606 TI - [73-year-old patient with hepatocellular carcinoma]. PMID- 19551605 TI - [Prolonged course of tick-borne ulceroglandular tularemia in a 20-year-old patient in Germany--case report and review of the literature]. AB - HISTORY AND ADMISSION FINDINGS: A 20-year-old female patient presented with painful axillary lymphadenopathy. She reported a tick bite five months ago in her right hand followed by fever, chills and regional lymphadenopathy. Empiric antibiotic treatment with doxycyclin and ciprofloxacin had led to defervescence but no change in painful lymph node swellings. Surgical removal of a cubital lymph node had already been performed three months after the tick bite. INVESTIGATIONS: Laboratory findings were normal except for moderate elevation of C-reactive protein. Serology confirmed the suspected clinical diagnosis of ulceroglandular tularemia. Retrospective real-time PCR (markers fopA and tul4) for Francisella tularensis from the previously removed lymph node (paraffin tissue blocks) was negative. DIAGNOSIS, TREATMENT AND COURSE: Clinical presentation and serological test results were consistent with a prolonged course of tick-borne ulceroglandular tularemia associated with reactive lymph node swelling. The patient requested surgical removal of the painful axillary lymph node. Histology showed reticulocytic, abscess forming lymphadenitis with pseudotuberculosis type of granulomatosis and negative acid-fast staining. Blood culture, capture ELISA and real-time PCR for Francisella tularensis performed on material from the lymph node preparations were negative. A complete recovery was achieved without renewed antibiotic treatment. CONCLUSIONS: According to recent seroprevalence studies, the emergence of tularemia as a rare zoonosis in Germany is clinically underestimated. This case report illustrates possible appearance of the disease in other than known risk groups (e.g. hunters, lumbermen). Ectoparasites like infected ticks have to be considered as vectors, even in non endemic regions. PMID- 19551608 TI - [Risk factors for sporadic Creutzfeldt-Jakob disease]. AB - Sporadic Creutzfeldt-Jakob disease (sCJD) is the most common form of human transmissible spongiform encephalopathies (prion disease), but its cause has not been fully elucidated. According to its biochemical properties prion protein is resistant to routine sterilisation methods. Thus, invasive medical procedures could be involved in the genesis of the disease. Present knowledge about iatrogenic routes of transmission, oral infection and transmission via blood products in variant CJD (vCJD) underlines the importance of careful surveillance and analysis of potential routes of transmission. Several studies of risk factors for sCJD published in the past have given contrary results, which may be largely explained by different control groups. This article reviews epidemiology and classification of CJD and discusses possible risk factors and summarizes previous case-control studies. PMID- 19551607 TI - [The problems of compliance and adherence, using the example of chronic inflammatory bowel disease]. AB - To what extent do patients comply with their doctor's treatment recommendation, i.e. show "compliance" and "adherence" ?. The treatment of chronic diseases in particular is associated with enormous problems of adherence. Patients often take only 40 to 80% of their prescribed dosage of medication. This issue affects not only the course of the disease in the particular patient, but also has considerable impact on the health care system. Great difficulties still exist in recognizing poor adherence, because doctors depend on the information given by their patients. The reasons for adherence problems can be attributed to disease related, treatment-related or patient-related factors, as well as to the quality of the doctor-patient relationship. Improving adherence is difficult due to its numerous and individually different causes. This has been emphasized by a recent Cochrane review, in which only 5 out of 21 randomized and controlled studies concerning the improvement of adherence demonstrated significant success. However, half of the studies displayed methodological errors that weakened the statistical detection of improvement because of the low numbers patients. Overall, enormous efforts will be required of doctors, patients and health care policies to achieve substantial alterations in the problems associated with adherence. PMID- 19551609 TI - RET expression and neuron-like differentiation of pheochromocytoma and normal chromaffin cells. AB - Receptor tyrosine kinase RET is normally expressed at low levels in chromaffin cells and high levels in sympathetic neurons. Paradoxically, it is overexpressed in subsets of pheochromocytomas. The overexpressed protein is usually wild-type, except in multiple endocrine neoplasia type 2. Possible explanations for overexpression include tumor origin from RET-expressing sympathoadrenal progenitors that escape developmental culling during embryogenesis, or reactivation of signaling pathways related to neuronal differentiation. Normal adult chromaffin and pheochromocytoma cells can undergo neuron-like differentiation in cell culture. In this investigation, cultured cells from two normal human adrenal medullas, two of three human pheochromocytomas, and one extra-adrenal paraganglioma showed RET induction corresponding with extensive nerve growth factor-induced outgrowth of neurite-like processes, while one pheochromocytoma showed neither processes nor RET induction. RET was similarly upregulated in parallel with process outgrowth in cultures of normal rat chromaffin cells and PC12 rat pheochromocytoma cells. In contrast, mouse pheochromocytoma cells that constitutively express high levels of wild-type RET together with other neuronal progenitor markers showed no further RET increase after cyclic AMP-induced process outgrowth. The RET-activating ligand GDNF was anti-apoptotic for mouse pheochromocytoma but not for PC12 cells. The findings suggest that overexpression of RET in pheochromocytomas could result either from a secondary event that activates signaling pathways mediating adult chromaffin cell plasticity or as a component of a persistent sympathoadrenal progenitor phenotype. Whether wild-type RET contributes to tumor development or is merely a lineage marker for cells at various stages of neuronal differentiation may vary, with other tumor characteristics. PMID- 19551610 TI - Prolactin suppresses malonyl-CoA concentration in human adipose tissue. AB - Prolactin is best known for its involvement in lactation, where it regulates mechanisms that supply nutrients for milk production. In individuals with pathological hyperprolactinemia, glucose and fat homeostasis have been reported to be negatively influenced. It is not previously known, however, whether prolactin regulates lipogenesis in human adipose tissue. The aim of this study was to investigate the effect of prolactin on lipogenesis in human adipose tissue in vitro. Prolactin decreased the concentration of malonyl-CoA, the product of the first committed step in lipogenesis, to 77+/-6% compared to control 100+/-5% (p=0.022) in cultured human adipose tissue. In addition, prolactin was found to decrease glucose transporter 4 ( GLUT4) mRNA expression, which may cause decreased glucose uptake. In conclusion, we propose that prolactin decreases lipogenesis in human adipose tissue as a consequence of suppressed malonyl-CoA concentration in parallel with decreased GLUT-4 expression. In the lactating woman, this regulation in adipose tissue may enhance the provision of nutrients for the infant instead of nutrients being stored in adipose tissue. In hyperprolactinemic individuals, a suppressed lipogenesis could contribute to an insulin resistant state with consequences for the health. PMID- 19551611 TI - Cytotoxic activity and cell cycle analysis of quinoline alkaloids isolated from Haplophyllum canaliculatum Boiss. AB - Bioassay-guided fractionation of Haplophyllum canaliculatum Boiss. (Rutaceae) extract resulted in isolation of five quinoline alkaloids: 7-isopentenyloxy-gamma fagarine, atanine, skimmianine, flindersine and perfamine. This is the first isolation of these compounds from this endemic species. The antitumor activity of these five isolates was evaluated against RAJI, Jurkat, KG-1a, HEP-2, MCF-7, HL 60 and HL-60/MX1 tumor cell lines. The highest cytotoxic effect was observed on acute lymphoblastic leukemia cell lines. 7-Isopentenyloxy-gamma-fagarine, atanine, skimmianine and flindersine exhibited very high cytotoxicity against the RAJI cell line with IC(50) values of 1.5, 14.5, 15.6 and 14.9 microg/mL, respectively and 7-isopentenyloxy-gamma-fagarine, atanine and skimmianine exhibited very high cytotoxicity against the Jurkat cell line with IC(50) values of 3.6, 9.3 and 11.5 microg/mL, respectively. 7-Isopentenyloxy-gamma-fagarine was also highly cytotoxic against the MCF-7 cell line (IC(50) = 15.5 microg/mL), while atanine, skimmianine, flindersine and perfamine showed moderate to low activity against these cells. All alkaloids had moderate to low cytotoxicity against KG-1a and HEP-2. Investigation of the toxic potential of the alkaloids on HL-60 and HL-60/MX1 showed a significantly higher effect against HL-60/MX1, a multidrug-resistant cell line, compared with the control etoposide (p < 0.05). In all cytotoxicity experiments, peripheral blood mononuclear cells (PBMC) were used as a control for normal hematopoietic cells. Flow cytometry analysis of the compounds resulted in the arrest of cell cycle progression at the sub-G1 phase of the RAJI and Jurkat cell lines in a dose-dependent manner. According to computational analyses, the similar cytotoxic trend in the cell lines could be indicative of the fact that these compounds may act through parallel mechanisms. PMID- 19551612 TI - Isolation and identification of novel genes involved in artemisinin production from flowers of Artemisia annua using suppression subtractive hybridization and metabolite analysis. AB - Malaria is a global health problem that threatens 300-500 million people and kills more than one million people annually. Artemisinin is highly effective against multidrug-resistant Plasmodium falciparum and it has been widely used as part of the artemisinin-based combination therapies against malaria. To elucidate the biosynthetic pathway of artemisinin and to clone related genes in Artemisia annua, differentially expressed genes between blooming flowers and flower buds were isolated and characterized by a combined approach of suppression subtractive hybridization (SSH) and metabolite analysis. A total of 350 cDNA clones from a subtractive cDNA library were randomly picked, sequenced and analyzed and 253 high-quality sequences were obtained. BLASTX comparisons indicated that about 9.9 % of the clones encoded enzymes involved in isoprenoid (including artemisinin) biosynthesis. The expression of 4 gene transcripts involved in artemisinin biosynthesis was examined by RT-PCR and the results confirmed the higher expression of these transcripts in blooming flowers than in flower buds. In addition, 2 putative transcript factors transparenta testa glabra 1 (TTG1) and ENHANCER OF GLABRA3 (GL3), which promote trichome initiation, were presented in the library. Finally, this study demonstrated that the increase of expression level of the putative TTG1 gene correlated with the improvement of glandular trichome density and artemisinin production in A. annua leaves. The subtractive cDNA library described in the present study provides important candidate genes for future research in order to increase the artemisinin content in A. annua. PMID- 19551613 TI - Overexpression of the HMG-CoA reductase gene leads to enhanced artemisinin biosynthesis in transgenic Artemisia annua plants. AB - An effective and affordable treatment against malaria is still a challenge for medicine. Most contemporary drugs either are too expensive to produce or are not effective against resistant strains of the malaria parasite Plasmodium falciparum. The plant Artemisia annua L. is the source of artemisinin, an effective drug against malaria for which no resistant strains of the bacterium have been reported. However, the artemisinin content of A. annua is very low, which makes its production expensive. Here we report the use of transgenic technology to increase the artemisinin content of A. annua. We report the production of transgenic plants of A. annua into which we transferred 3-hydroxy-3 methylglutaryl CoA reductase (HMGR) gene from Catharanthus roseus (L.) G. Don using Agrobacterium-mediated gene transfer technology. Transgene integration and copy number were assessed by PCR and Southern hybridization, which confirmed the stable integration of multiple copies of the transgene in 7 different transgenic lines of A. annua. The leaf tissue of three of the A. annua transgenic lines possessed significantly higher HMGR activity compared with wild-type controls, and this activity was associated exclusively with microsomal membranes. The artemisinin content of the shoots of one of the transgenic lines depicted an increase of 22.5 % artemisinin content compared with wild-type control A. annua plants. PMID- 19551614 TI - Ultrastructural changes on clinical isolates of Trichophyton rubrum, Trichophyton mentagrophytes, and Microsporum gypseum caused by Solanum chrysotrichum saponin SC-2. AB - Worldwide, dermatophytoses represent a high percentage of all superficial mycoses. The most frequently isolated dermatophyte is Trichophyton rubrum. Solanum chrysotrichum is a vegetal species widely used in Mexican traditional medicine to treat skin infections; its extract has been used to formulate an herbal medicinal product that is used successfully to treat Tinea pedis. Spirostanic saponin SC-2 from S. Chrysotrichum possesses high activity against dermatophytes. The present study reports the ultrastructural changes observed by means of transmission electron microscopy (TEM) in clinical isolates of T. rubrum, T. mentagrophytes, and Microsporum gypseum induced by saponin SC-2. Strains were grown in RPMI 1640 containing SC-2 (1600 microg/mL). Fungi were harvested at 6, 12, 24, and 48 h; controls without SC-2 were included. T. mentagrophytes was the most susceptible to the SC-2 saponin, followed by M. gypseum, while T. rubrum was the most resistant. The main alterations caused by the SC-2 saponin were as follows: i) loss of cytoplasmic membrane continuity; ii) organelle degradation; iii) to a lesser extent, irreversible damage to the fungal wall; and iv) cellular death. PMID- 19551616 TI - Incomplete tubular duplication of the esophagus becoming symptomatic in adulthood. PMID- 19551615 TI - Reduction of postoperative adhesions by perfluorocarbons: an experimental study in a rat model. AB - Peritoneal adhesions are a well-known and frequently occurring postoperative complication. Many published studies have looked into the prophylaxis of adhesions following abdominal surgery, but only few clinically relevant agents have been reported. Most publications refer to adult patients and not to paediatric patient collectives. This experimental study in a rat model compares the effect of perfluorocarbons as adhesion prophylaxis with those of a well-known anti-adhesive agent Adept and with an untreated control group. We hypothesized that PFC might have a double effect: initially it could suppress the accumulation of monocytes and neutrophilic granulocytes, and subsequently it would work as a barrier to prevent contact between the visceral and parietal layers of the peritoneum. After a standardised operation, PFC was injected into the abdominal cavity of rats in the study group. Macroscopically, the PFC group did not fare significantly better, but nevertheless a clear tendency towards fewer adhesions after the application of PFC could be ascertained. PMID- 19551617 TI - Puncture and cytology - sufficient for endoscopic diagnosis of pneumatosis cystoides intestinalis? PMID- 19551618 TI - [Pressure ulcer prevalence in German nursing homes and hospitals: what role does the National Nursing Expert Standard Prevention of Pressure Ulcer play?]. AB - AIM OF THE STUDY: The aim of this study was to investigate the relationship between the use of the National Nursing Expert Standard Pressure Ulcer Prevention and the pressure ulcer prevalence in German nursing homes and hospitals. METHODS: Data were collected within two nationwide surveys conducted by the Department of Nursing Science of the Charite, Berlin, Germany. The surveys, designed as cross sectional prevalence studies, serve as an investigation of the amount of clinically relevant nursing phenomena, i. e., pressure ulcers. Prevalence per facility in the at-risk group was explored by a ranking procedure of the 95 nursing homes and hospitals. The facilities were divided into two groups according to whether they used the German Expert Standard to develop the local protocol or not. RESULTS: The pressure ulcer prevalence of the at-risk group ranged from 0% to 24.6% in nursing homes and from 7% to 40% in hospitals. In about 40% of the hospitals and nursing homes the local protocol of pressure ulcer prevention was based on the German Expert Standard. The ranking figure indicates that there is no statistically significant relation between Expert Standard-based local protocols and the pressure ulcer prevalence in the at-risk group. CONCLUSION: A clear advantage to use the German Expert Standard compared with other sources cannot be shown with these data. However, a uniform pressure ulcer prevention is an essential quality feature of nursing care. The degree of implementation and the consequent transfer of the recommendations to daily practice should be evaluated regularly. PMID- 19551619 TI - [Medical services: patients' and physicians' perceptions]. AB - In 2007, the German health-care system was once again subject to reform. Demographic changes and a rising demand for health-care services on the one hand and technological progress with increased medical-technological possibilities on the other lead to questions about organising and financing health care. Moreover, with the German health care being a mixture of a public insurance systems and private insurances, the extent, quality and equality of access is constantly under debate. In an empirical study, 1 604 patients were asked about their satisfaction with medical services and their insurance as well as their views on the recent reform. 90 physicians were questioned about their job satisfaction and their views on the health care system and the recent reform. Patients, irrespective of their type of insurance, on average were very satisfied with the physicians' services. Privately insured patients on average were more satisfied with their insurance than publicly insured. Patients were not well informed about the reform. Physicians' job satisfaction was markedly lower, they anticipate problems with health-care provision and they judge the health-care reform as inappropriate. Thus, patients on average are very satisfied with medical services, physicians on average are not satisfied with their situation, and the health-care reform is not perceived as a great success. PMID- 19551620 TI - [Evaluation of empowerment among socially disadvantaged women - examination in different living circumstances]. AB - This paper follows on from a previous study which assessed the relationship between socioeconomic position, empowerment and the development of psychological health in women after treatment in mother-child rehabilitation centres in Germany. The study revealed that socioeconomic position was less important for mothers caring for young children. For this reason the present study is based on a broader definition of social inequity, taking household conditions and psychosocial stressors into account. The aim of the paper is to answer the following questions: 1) To what extent does the improvement of psychological health depend on the living circumstances of the mothers? 2) What is the impact of living conditions on the success of empowerment? 3) Does the health-related impact of empowerment differ between different living conditions of the mothers? By conducting a cluster analysis on clinical data of the women (n=6094), seven different living circumstances of the mothers could be detected. Two living circumstances could be identified to be related to extremely poor health. These are 'dissatisfied single mothers with high degrees of psychosocial distress and lack of social support', and 'married mothers with conflicts within the family and self-perceived lack of appreciation'. At the end of inpatient treatment these mothers showed the highest reduction of psychological symptoms, but after six and twelve months the symptoms increased again. The results of empowerment showed that empowerment is most health-effective for mothers living in poor living conditions, but the success of empowerment here is less pronounced. As a consequence the health effect of empowerment was smaller for those mothers. The study suggests that health promotion programmes could be more effective when they explicitly take the living circumstances of their participants into account. PMID- 19551621 TI - [Radiation risks from diagnostic radiology: meningiomas and other late effects after exposure of the skull]. AB - A complete assessment of late effects of X-ray diagnostics should take into account that radiation sensitivity varies considerably for the different ages at exposure and, furthermore, that not only malignant diseases but also benign neoplasms are induced which also may lead to severe detriment of the patient. Risk estimates are derived for paediatric head CTs as well as for brain tumours in adults. Dose-effect relationships for tumours of the brain, skin, thyroid, and other sites of the head region, leukaemia, and cataracts are taken from the literature. On the basis of estimates for Germany about the number of head scans, the annual rate of radiation-induced diseases is calculated. 1,000 annual paediatric CT investigations of the skull will lead to about 3 excess neoplasms in the head region, i.e., the probability of an induced late effect must be suspected in the range of some thousandths. Additionally, a relevant increase of cataracts must be considered. The radiation-induced occurrence of meningiomas and other brain tumours most probably contributes to the continuously increasing incidence of these diseases which is observed in several industrial nations, as well as the exposure of the bone marrow by CT to the increase of childhood leukaemia. PMID- 19551622 TI - [Decision-analytical modelling of costs per QALY in the context of the German Social Law]. AB - PURPOSE: In 2007, a legal reform introduced formal health economic evaluation for selected reimbursement decisions by the statutory health insurance in Germany. The methods of evaluation are currently under discussion. This study assesses whether an approach based on decision-analytic cost per QALY modelling fits with the legal requirements set by Book Five of the German Social Code (SGB V). METHODS: It is based on a review of legal documents and the relevant literature. RESULTS: Key specifications for economic evaluation in Germany are the differential interpretations of "benefit" in the relevant legislation as well as the requirement that the methods follow "international standards of evidence based medicine and health economics" ( section sign section sign 35b, 139a SGB V). In German reimbursement decision practice, new interventions have undergone an assessment of (1) benefit, (2) necessity and (3) cost-effectiveness (prior to the legal reform only exclusion of dominated alternatives). While the establishment of benefit in step (1) is preferably based on clinical trials in current practice, also two different interpretations of "benefit" in steps (1) and (3) would be in accord with SGB V. Methods for establishing QALYs measure and evaluate different dimensions of health benefit based on transparent and theoretically justified methods. They also capture the dimensions specifically stated by section sign 35b SGB V, e. g., extension of life or improvement in quality of life. Compared to ad-hoc synthesis of the different measures of patient benefit from clinical trials, this is more consistent with the standards of evidence-based medicine and health economics. Decision analytical modelling provides a practical and theoretically sound method to integrate the condition specific evidence for estimating the costs and benefits associated with a medical treatment in health care practice. CONCLUSIONS: Both the establishment of health effects in terms of QALYs and decision-analytical models for evidence synthesis meet the requirements of SGB V. Further methodological issues that need to be addressed include guidelines for QALY measures, and the choice of analytical perspective that fits best with German law. This notwithstanding, it remains an open question how the appropriateness of a cost-benefit ratio should be appraised and what role cost-effectiveness should play in health care decision making, compared with other principles potentially relevant to the decision. PMID- 19551623 TI - [Unspecific back pain - basic principles and possibilities for intervention from a psychological point of view]. AB - INTRODUCTION: Back pain is one of the most common and expensive health problems in Germany. Apart from somatic parameters, psychological factors are thought to influence the aetiology and, especially, the chronification of back pain. A literature search has been performed to establish which psychological procedures are used in the diagnosis, prevention and therapy for back pain, and how effective they are. METHOD: The connection between back pain and psychological factors was investigated in English and German technical articles identified in a literature search in Medline (2000-2006) and in psyndex (2000-2006). 714 hits were identified for the key words "back pain and prevention", 61 hits for "back pain and psychological factors", 732 hits for "back pain and risk factors" and 4 hits for "back pain and chronification of pain". 75 technical articles or studies were selected from these hits and used as a basis for the evaluation of the above questions. The search was completed by a manual search in the literature lists of published articles, perusing monographs, searching the internet and evaluating congress and meeting reports (2000-2008). RESULTS: The results of various studies show that cognitive, emotional and behavioural aspects and the processes of respondent and operant learning are of great importance in processing and overcoming pain. Biopsychosocial procedures appear to be superior to biomedical procedures. There has been good research on the (psychological) risk factors which contribute to the chronification of pain. It seems to be sensible to use screening procedures, such as the "Orebro Musculoskeletal Pain Questionnaire (OMPQ)" for the identification of patients at increased risk of chronification. For patients with chronic symptoms, there are evidence-based psychological programme components; these are usually successful, especially as part of multimodal programmes. As yet, there have only been initial studies on the use of psychological intervention for prevention, particularly in occupational settings. DISCUSSION: In the high risk group of patients already suffering from back pain, relapses and chronification can best be prevented by multimodal programmes. A decisive condition for the success of these physiotherapeutic, ergotherapeutic, sports therapeutic and psychotherapeutic interventions is apparently that there should be a standardised procedure in accordance with the theoretical principles of behavioural therapy, complied with by all those involved in the process. However, the principle objective of all measures should be the avoidance of the transition from acute to chronic back pain and the protracted and expensive clinical course this can lead to. Psychological intervention appears to work here as well. Further studies are needed to clarify whether the psychological program components are effective in isolation or whether they must be embedded in a multimodal (preventive) concept. It must also be investigated which target groups benefit most from which type of preventive (psychological) intervention. Provision of specific information is an alternative preventive approach. This health psychological or educative procedure corresponds to demedicalisation of this condition. Although media campaigns have been successfully performed in Australia, it is as yet unclear whether these can be transferred to Germany and which structural changes these would require in our health care system. PMID- 19551624 TI - [Cost-effectiveness model of a community-based service for dementia caregivers]. AB - OBJECTIVE: The objective of this study was to compare the cost-effectiveness of a psychoeducational concept for dementia caregivers with standard care. METHODS: A literature search determined the data for the cost-effectiveness model concerning the quality-adjusted life years (QALYs) and costs. The parameters were evaluated using a decision tree. The evaluation of the base case analysis concerning stability was carried out using sensitivity analysis and a Monte Carlo simulation (MCS). RESULTS: After the implementation of the psychoeducational concept for caregivers a saving of 750 euro was made and a 0.07 QALY was gained over a one year time horizon. The results of the sensitivity analysis proved to be stable. CONCLUSIONS: By using the community-based service, costs can be saved and QALYs of the caregivers can be gained. This results in a decrease of depressive symptoms among caregivers and in a delay of institutionalisation of the dementia patient. PMID- 19551625 TI - [The term "Funktionale Gesundheit" (functional health) in the German edition of the ICF]. AB - In the preface of the German edition of the INTERNATIONAL CLASSIFICATION OF FUNCTIONING, DISABILITY AND HEALTH (ICF) the term "functional health" (Funktionale Gesundheit) is introduced and defined. There is no direct equivalent expression in the original English Version of the ICF. The definition of "Funktionale Gesundheit" was thus strongly disputed among the translators and ICF experts involved in the German translation. In the following, the author suggests reconsidering the definition critically and proposes an alternative solution. PMID- 19551626 TI - [Prevalence and utilisation of health promotion in German enterprises]. AB - The aim of this study was to estimate the prevalence of health promotion in German enterprises, differentiated by size, sector, and the companies' business situations. Representative data were analysed from the survey of working conditions in Germany (n=20,000) that was conducted by the Federal Institute for Vocational Education and Training (BIBB) and the Federal Institute of Occupational Safety and Health (BAuA). Thirty-eight percent of interviewed employees confirmed that health promotion activities had been carried out in their company during the last two years. The prevalence varied significantly, depending on the company's size, sector, and business situation. In cases of implementation, a higher percentage of employees participated in micro and small companies than in medium-sized or large companies. With respect to the implementation of health promotion, more advice and support are needed, particularly in micro and small enterprises. There is still a need for health promotion activities which meet the special needs of micro and small enterprises. Furthermore there is still a need to invest in an infrastructure which allows their adequate supply. PMID- 19551627 TI - Molecular characterization of olive cultivars using amplified fragment length polymorphism markers. AB - We sampled six olive cultivars (Tavli Sati, Sati, Gorvela, Sacakli Otur, Butko, and Otur) from Coruh Valley, located in the northeast part of Turkey, and characterized them using amplified fragment length polymorphism (AFLP) markers. Some morphological and biochemical characteristics are also determined. Six AFLP primer combinations were used for molecular characterization and 66 AFLP markers were obtained. Six olive cultivars were classified into two major clusters using UPGMA clustering analysis; cv. Otur alone comprised the first group. Some morphological and biochemical characteristics of cv. Otur were also distinct from those of the other cultivars. The highest genetic similarity was observed between cultivars Tavli Sati and Sati (0.74), while the lowest genetic similarity was observed between cvs. Gorvela and Otur (0.37). PMID- 19551628 TI - Identification of Turkish and standard apple rootstocks by morphological and molecular markers. AB - Two local (Vezir-1 and Vezir-2) and two standard (M9 and MM106) clonal apple rootstocks were compared using both morphological and molecular markers. International Union for the Protection of New Varieties of Plants criteria were used for morphological evaluation, which did not clearly separate these rootstocks. We tested 47 random decamer primers for random amplified polymorphic DNA analysis; 15 of them gave reproducible polymorphic patterns, yielding 109 bands, which showed 78% polymorphism. Based on a dendrogram obtained by unweighted pair group method using arithmetic average analysis, three clusters were obtained. The highest genetic similarities were found between M9 and Vezir-2 (0.670). The random amplified polymorphic DNA markers proved to be more efficient than the standard morphological markers for the identification of rootstocks. PMID- 19551629 TI - A patient with ascending aortic dilatation, similar to phenotypes of connective tissue disorders. AB - We report on the clinical and molecular findings of a patient who presented alopecia, epicanthus, micrognathia, retrognathia, high arched palate, hypertelorism, Chiari type I malformation, mixed-type hearing loss but with normal heartbeat Q-T interval, malformed earlobes, down-slanted palpebral fissures, downturned corners of the mouth, syndactyly, atopic eczema, and seizures. The patient was a male adult, 23 years old, with short stature (153 cm) and low weight (50.5 kg), due to severe aortic insufficiency and dilatation of the ascending aorta. Conventional cytogenetic screening did not show any chromosomal gains or losses. Molecular genetic screening was conducted for gene mutations involved in various syndromes; the mutations found included [beta fibrinogen -455 G>A wt/wt (wt/mut), PAI-1 4G/5G (4G/4G), HPA1 a/b (a/a), MTHFR C677T wt/wt (wt/mut), ACE I/D (I/I), and Apo E E3/E4]. Many clinical and molecular genetics findings overlapped with other conditions associated with arterial tortuosity and arterial aneurysms, including the Marfan, Ehler-Danlos, Shprintzen-Goldberg, and Loeys-Dietz syndromes. Although a diagnosis of Shprintzen-Goldberg syndrome was based on clinical findings and radiographic findings indicate other syndromes, aortic root dilatation seems to be a new symptom, similar to phenotypes of connective tissue disorders. The unique grouping of clinical manifestations in this patient and the molecular genetics findings lead us to suggest that this case could be an example of a previously unrecognized syndrome. PMID- 19551630 TI - The G1138A mutation rate in the fibroblast growth factor receptor 3 (FGFR3) gene is increased in cells carrying the t (4; 14) translocation. AB - Spontaneous mutations are a common phenomenon, occurring in both germ-line and somatic genomes. They may have deleterious consequences including the development of genetic disorders or, when occurring in somatic tissues, may participate in the process of carcinogenesis. Similar to many mutational hotspots, the G1138A mutation in the fibroblast growth factor receptor 3 (FGFR3) gene occurs at a CpG site. In germ-line tissues, the G1138A mutation results in achondroplasia and has one of the highest spontaneous mutation rates in the human genome. Although not at the G1138A site, there are increased rates of other somatic mutations in the FGFR3 gene that have been reported in multiple myeloma cases associated with a translocation, t (4; 14). The chromosome-4 break points in this translocation are clustered in a 70-kb region centromeric to the FGFR3 gene. We hypothesized that this translocation may impact the mutation rate at the G1138A site. We employed a semi-quantitative polymerase chain reaction-based assay to measure the frequency of this mutation in multiple myeloma cell lines carrying t (4; 14) translocation. Analysis of these cell lines varied from no change to a 10-fold increase in the mutation frequency compared with normal controls. In general, there was an increase in the G1138A mutational frequency suggesting that chromosomal rearrangement can affect the stability of the CpG hotspots. PMID- 19551631 TI - Genotoxicity testing of Ambelania occidentalis (Apocynaceae) leaf extract in vivo. AB - Ambelania occidentalis is routinely used in folk medicine for treating gastrointestinal disorders, even though there have been no safety trials. We evaluated the genotoxic potential of hydro-alcoholic extracts of this plant in mice; induced DNA damage was assessed in peripheral blood leukocytes and micronucleus induction was assessed in polychromatic erythrocytes from bone marrow. The extract was administered by an oral route at single doses of 1000, 1500 and 2000 mg/kg body weight. N-nitroso-N-ethylurea was used as a positive control. The comet assay was performed on peripheral blood leukocytes at 4 and 24 h after treatment, and the micronucleus test was carried out on bone marrow cells collected at 24 and 48 h after treatment. The ratio of polychromatic/normochromatic erythrocytes was scored for cytotoxicity assessment. No increase in the number of micronucleated polychromatic erythrocytes from bone marrow or in leukocyte DNA damage was observed. The hydro-alcoholic extracts of A. occidentalis had no mutagenic or cytotoxic effects in the mouse cells. PMID- 19551632 TI - A proposed selection index for feedlot profitability based on estimated breeding values. AB - It is generally accepted that feed intake and growth (gain) are the most important economic components when calculating profitability in a growth test or feedlot. We developed a single post-weaning growth (feedlot) index based on the economic values of different components. Variance components, heritabilities and genetic correlations for and between initial weight (IW), final weight (FW), feed intake (FI), and shoulder height (SHD) were estimated by multitrait restricted maximum likelihood procedures. The estimated breeding values (EBVs) and the economic values for IW, FW and FI were used in a selection index to estimate a post-weaning or feedlot profitability value. Heritabilities for IW, FW, FI, and SHD were 0.41, 0.40, 0.33, and 0.51, respectively. The highest genetic correlations were 0.78 (between IW and FW) and 0.70 (between FI and FW). EBVs were used in a selection index to calculate a single economical value for each animal. This economic value is an indication of the gross profitability value or the gross test value (GTV) of the animal in a post-weaning growth test. GTVs varied between -R192.17 and R231.38 with an average of R9.31 and a standard deviation of R39.96. The Pearson correlations between EBVs (for production and efficiency traits) and GTV ranged from -0.51 to 0.68. The lowest correlation (closest to zero) was 0.26 between the Kleiber ratio and GTV. Correlations of 0.68 and -0.51 were estimated between average daily gain and GTV and feed conversion ratio and GTV, respectively. These results showed that it is possible to select for GTV. The selection index can benefit feedlotting in selecting offspring of bulls with high GTVs to maximize profitability. PMID- 19551633 TI - Haplotype distribution of five nuclear genes based on network genealogies and Bayesian inference indicates that Trypanosoma cruzi hybrid strains are polyphyletic. AB - Chagas disease is still a major public health problem in Latin America. Its causative agent, Trypanosoma cruzi, can be typed into three major groups, T. cruzi I, T. cruzi II and hybrids. These groups each have specific genetic characteristics and epidemiological distributions. Several highly virulent strains are found in the hybrid group; their origin is still a matter of debate. The null hypothesis is that the hybrids are of polyphyletic origin, evolving independently from various hybridization events. The alternative hypothesis is that all extant hybrid strains originated from a single hybridization event. We sequenced both alleles of genes encoding EF-1alpha, actin and SSU rDNA of 26 T. cruzi strains and DHFR-TS and TR of 12 strains. This information was used for network genealogy analysis and Bayesian phylogenies. We found T. cruzi I and T. cruzi II to be monophyletic and that all hybrids had different combinations of T. cruzi I and T. cruzi II haplotypes plus hybrid-specific haplotypes. Bootstrap values (networks) and posterior probabilities (Bayesian phylogenies) of clades supporting the monophyly of hybrids were far below the 95% confidence interval, indicating that the hybrid group is polyphyletic. We hypothesize that T. cruzi I and T. cruzi II are two different species and that the hybrids are extant representatives of independent events of genome hybridization, which sporadically have sufficient fitness to impact on the epidemiology of Chagas disease. PMID- 19551634 TI - Alpha-thalassemia (3.7 kb deletion) in a population from the Brazilian Amazon region: Santarem, Para State. AB - The ethnic composition of the Brazilian population favors high frequencies of the -alpha3.7 deletion, responsible for alpha-thalassemia, because this mutation is very common in African populations. In spite of its importance, this hemoglobinopathy has been poorly investigated in Brazil, especially at the molecular level. We investigated the prevalence of the -alpha3.7 mutation in 220 individuals attended at the Municipal Hospital of Santarem, in the state of Para. These patients were distributed into three different groups: i) 103 individuals with anemia who had microcytosis and hypochromia, ii) 11 individuals without anemia who had microcytosis and hypochromia, and iii) 106 individuals with no hematological alterations. We examined the usefulness of investigating alpha thalassemia carrier status for microcytosis. Among the 103 patients with anemia, 20 (19.4%) were heterozygotes (-alpha3.7/alphaalpha) and one (1.0%) was a homozygote (-alpha3.7/-alpha3.7). Among the 11 patients without anemia, one heterozygote (-alpha3.7/alphaalpha) was identified; in the third group, composed of normal individuals (106 samples), deletion -alpha3.7 was found in seven samples (6.6%), all of which were heterozygotes (-alpha/alphaalpha).These frequencies are within the expected range, given available data on the distribution of this hemoglobin disorder in human populations and the ethnic composition of the population of Santarem. We found that alpha-thalassemia is a common cause of microcytosis, given that a high proportion (19.2%) of the microcytic population carried alpha-globin gene deletions. PMID- 19551635 TI - A rapid and efficient method for isolation of total RNA from Euglena gracilis (Euglenoidea). AB - RNA isolation is essential to the study of gene expression at the molecular level. However, it is difficult to isolate RNA from organisms that contain large amounts of polysaccharides or other compounds that bind or coprecipitate with RNA, such as the unicellular protist Euglena gracilis. Currently, there is no commercial kit available that is specific for the isolation of high-quality RNA from this organism. Since it contains large amount of polysaccharides, the common protocols for RNA isolation usually result in poor yields when applied to E. gracilis. We developed a simple and fast RNA protocol that effectively removes these contaminating substances, without affecting the RNA yield. This protocol was based on the sodium dodecyl sulfate/phenol method, without beta mercaptoethanol and without maceration in liquid nitrogen; it uses phenol/chloroform extraction to remove proteins, DNA, and co-precipitated polysaccharides. The RNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder RNA extraction. PMID- 19551636 TI - A985G mutation incidence in the medium-chain acyl-CoA dehydrogenase (MCAD) gene in Brazil. AB - In view of the serious consequences of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency and the absence of information about its incidence in the Brazilian population, we examined the frequency of the A985G mutation in the MCAD gene. A retrospective analysis was made of data on 1722 individuals (844 females) genotyped for the A985G mutation in the MCAD gene, using genomic DNA extracted from peripheral blood leukocytes and genotyping with PCR-RFLP; 0.41% of these individuals were heterozygous for the A985G mutation. The mutant homozygous genotype was not found. The 985G mutant and 985A normal alleles had allelic frequencies of 0.0020 and 0.9980, respectively. Given the A985G allele frequency, genotyping would be recommended in cases of family history of MCAD deficiency and sudden infant death syndrome, and when there is suspicion of medium-chain fatty acid metabolic alterations; genetic counseling should be offered in cases involving 985GG and A985G individuals and consanguineous marriages. PMID- 19551637 TI - Analysis of p53 codon 72 gene polymorphism in Brazilian patients with endometriosis. AB - We examined the frequency of p53 codon 72 polymorphism in 38 patients with endometriosis whose diagnosis was confirmed using videolaparoscopy. Half of the women were infertile. There were no significant differences in the genotype (P = 0.0927) or allele frequencies (P = 0.1430) for p53 Arg72Pro polymorphism between the two groups. We found a significant association between the heterozygous and homozygous proline genotypes and intense pain in the patients. Sixty-four percent of the patients were homozygous or heterozygous for proline in patients with degree III or IV endometriosis, but there was no significant difference compared to homozygous arginine genotype (P = 0.6115). We found that the proline allele is associated with substantial complaints (infertility associated with pain), when compared to the homozygous arginine genotype; we also found that the proline allele was more frequent in endometriosis patients. PMID- 19551638 TI - Screening of mutations in the GCK gene in Jordanian maturity-onset diabetes of the young type 2 (MODY2) patients. AB - Maturity-onset diabetes of the young type 2 (MODY2) is a genetic form of diabetes mellitus caused by mutations in the glucokinase gene (GCK). We assessed the frequency of GCK gene mutations in Jordanian suspected MODY2 patients. We screened exons 7, 8 and 9, which are specific for pancreatic glucokinase, for mutations at positions 682A>G, p.T228A; 895G>C, p.G299R, and 1148C>A, p.S383X, respectively, in 250 subjects (100 patients suspected to have MODY2 and 150 healthy controls without family history of diabetes mellitus). We did not find any association of these mutations in Jordanian suspected MODY2 patients or in healthy controls, different from data on Caucasian Italian patients screened for the same mutations. PMID- 19551639 TI - A mixed colony of Scaptotrigona depilis and Nannotrigona testaceicornis (Hymenoptera, Apidae, Meliponina). AB - We describe a case of a spontaneously established mixed colony of two species of stingless bees. The host colony of Scaptotrigona depilis, an aggressive bee that forms large colonies, was invaded by workers of Nannotrigona testaceicornis, a smaller bee that forms small colonies. The host colony and the invading species colony were maintained in next boxes about 1.5 m apart. The N. testaceicornis colony had been recently divided. Observations were made daily for 10 min, and every two weeks the colony was opened for observations within the nest. Initially the host colony bees repulsed the invading species, but as their numbers built up, they were no longer able to defend the entrance. An estimated 60-90 N. testaceicornis workers lived integrated into the colony of S. depilis for 58 days. During this period, they reconstructed and maintained the entrance tube, changing it to an entrance typical of N. testaceicornis. They also collected food and building material for the host colony. Nannotrigona testaceicornis tolerated transit of S. depilis through the entrance, but did not allow the host species to remain within the tube, though the attacks never resulted in bee mortality. Aggression was limited to biting the wings; when the bees fell to the ground they immediately separated and flew back. There have been very few reports of spontaneously occurring mixed stingless bee colonies. It is difficult to determine what caused the association that we found; probably workers of N. testaceicornis got lost when we split their colony, and then they invaded the colony of S. depilis. PMID- 19551640 TI - Male sleeping aggregations of solitary oil-collecting bees in Brazil (Centridini, Tapinotaspidini, and Tetrapediini; Hymenoptera: Apidae). AB - Males of solitary bees usually spend the night in clusters on small branches of plants, cavities and flowers. The individuals usually return to the same location each evening during their life, exhibiting site fidelity to a particular plant. We report on the sleeping roosts of the males of some oil-collecting bees of the genera Centris, Paratetrapedia, Lanthanomelissa, Monoeca, and Tetrapedia, as well as the host plants. We discuss the role of the male clusters to the associated plants. PMID- 19551641 TI - Bionomics and sociological aspects of Euglossa fimbriata (Apidae, Euglossini). AB - The nesting biology and the social structure of Euglossa fimbriata were studied based on two original nests found on the campus of the University of Sao Paulo, Ribeirao Preto, Brazil. Nest 1 contained two inseminated females, 39 sealed cells, one cell being provisioned, and six old and empty cells. Nest 2 contained three inseminated females, 45 sealed cells, one cell being provisioned, and 27 old and empty cells. The cells of nest 1 were distributed into three clusters from which three new nests were set up in the laboratory and maintained in observation boxes from August 1993 to March 1994 in order to study the behaviors performed by the bees. The males left the nest immediately after emergence and did not return. Some females left the nest within a few days of eclosing, and others stayed in their original nests and began to reactivate them. The E. fimbriata colonies were small, with semi-social and eusocial organization. In these colonies one female becomes the dominant female, usually the oldest female, and the others behave as subordinate females. The subordinate females build their cells, provision and oviposit in them, while the dominant female becomes the major guard bee, and oviposits in cells oviposited in by subordinate females. Oviposition by the dominant female is always preceded by oophagy. Irrespective of the behavior displayed, all the females that we dissected had been inseminated. The behavior displayed by the dominant female is characteristic of brood parasitism and fits the parental parasitism hypothesis developed as an alternative pathway by which insect sociality could have arisen. PMID- 19551642 TI - Differences between the quality of strawberries (Fragaria x ananassa) pollinated by the stingless bees Scaptotrigona aff. depilis and Nannotrigona testaceicornis. AB - We investigated the success of two stingless bee species in pollinating strawberries in greenhouses. Three greenhouses and one open field area were used; one greenhouse had only strawberry plants (control), another (G1) had three colonies of Scaptotrigona aff. depilis and another (G2) had three colonies of Nannotrigona testaceicornis. In the open field area, the flowers could be visited by any bee. The total production of fruits was counted and a random sample (N = 100) from each area was used to measure weight, length, circumference, and achenes number (N = 5). The percentages of deformed strawberries were: 23% (no bees); 2% (greenhouses with bees) and 13% (open field). The strawberries from the greenhouse with N. testaceicornis and the open field were heavier than those from the greenhouses with no bees and with S. depilis. The fruit circumference was largest in the greenhouses with bees. The achenes number did not differ among the experimental areas. The strawberries produced in the greenhouses with stingless bees had more quality and greater commercial value than the fruits produced in the open field area and the greenhouse without bees. We conclude that stingless bees are efficient pollinators of strawberry flowers cultivated in greenhouses. PMID- 19551643 TI - Pollen sources of the orchid bee Euglossa annectans Dressler 1982 (Hymenoptera: Apidae, Euglossini) analyzed from larval provisions. AB - In order to analyze the pollen resources used by the orchid bee Euglossa annectans, samples of larval provisions from cells under construction were taken from 12 different trap nests (wooden boxes) on Santa Catarina Island, southern Brazil. The 43 samples collected between 2002 and 2005 represented all months except December. Overall, 74 pollen types from 24 families were distinguished. Among the 26 pollen types that reached more than 10% in monthly means, the families Melastomataceae, Bromeliaceae, Ochnaceae, Fabaceae, and Myrtaceae were most frequently represented. The Shannon-Weaver diversity index H' for the 43 brood cells varied from 0.10-1.65 and the annual diversity was 0.98. Similarity indices ranged from 0 to 0.87 and were highest during spring and summer. The results characterize E. annectans as a polylectic species. Based on these data, we can conclude that Euglossa females may act as pollinators of many forest species. PMID- 19551644 TI - The number of reproductive workers in highly eusocial Hymenoptera: monogyny and monandry. AB - Haplodiploidy results in relatedness asymmetries between colony members of highly eusocial Hymenoptera. As a consequence, queen and reproductive workers are more related to their own sons than to each other's male offspring. Kin selection theory predicts multiple optima in male parentage: either the queen or the workers should produce all the males. Nevertheless, shared male parentage is common in highly eusocial hymenopterans. An inclusive fitness model was used to analyze the effect of the number of reproductive workers on male parentage shared by the queen and laying workers by isolating the male component from an inclusive fitness equation using the equal fitness through male condition for each pairwise combination of the three female classes comprised of the queen, laying workers and non-laying workers. The main result of the theoretical analyses showed that the fraction of males produced by workers increases asymptotically with the number of laying workers at an increasingly diminishing rate, tending to an asymptotic value of 0.67. In addition, as the number of laying workers increases, the share of male parentage converges to that of non-laying workers. The diminishing return effect on male parentage share depending on the number of reproductive workers leads us to expect the number of reproductive workers to be relatively small in a stingless bee colony, even in the absence of productivity costs. The available data confirms this hypothesis, as there is an unusually small number of reproductive workers in stingless bee colonies. PMID- 19551645 TI - Hygienic behavior in the stingless bees Melipona beecheii and Scaptotrigona pectoralis (Hymenoptera: Meliponini). AB - Hygienic behavior, a trait that may confer resistance to brood diseases in the honey bee Apis mellifera, was studied in two species of stingless bees in Mexico. Eight colonies each of Melipona beecheii and Scaptotrigona pectoralis were tested for hygienic behavior, the removal of dead or diseased brood, by freeze killing a comb of sealed cells containing pupae. Both species detected and removed dead brood. However, removal rates differed between species. In M. beecheii colonies, workers took 2-9 days to remove 100% of the dead brood (4.4 +/- 2.0 days, mean +/ SD), while S. pectoralis removed all dead brood in less than 3 days (2.3 +/- 0.6 days, mean +/- SD). We conclude that hygienic behavior is not unique to A. mellifera, and is not solely an adaptation for the reuse of brood cells as occurs in honey bees but not stingless bees. Although stingless bees do not reuse brood cells, space is limited. The removal of dead brood may be necessary to allow new cells to be constructed in the same place. PMID- 19551646 TI - Spatial patterns in the brood combs of Nannotrigona testaceicornis (Hymenoptera: Meliponinae): male clusters. AB - Genetic models of sex and caste determination in eusocial stingless bees suggest specific patterns of male, worker and gyne cell distribution in the brood comb. Conflict between queen and laying workers over male parentage and center periphery gradients of conditions, such as food and temperature, could also contribute to non-random spatial configuration. We converted the positions of the hexagonal cells in a brood comb to Cartesian coordinates, labeled by sex or caste of the individuals inside. To detect and locate clustered patterns, the mapped brood combs were evaluated by indexes of dispersion (MMC, mean distance of cells of a given category from their centroid) and eccentricity (DMB, distance between this centroid and the overall brood comb centroid) that we developed. After randomizing the labels and recalculating the indexes, we calculated probabilities that the original values had been generated by chance. We created sets of binary brood combs in which males were aggregated, regularly or randomly distributed among females. These stylized maps were used to describe the power of MMC and DMB, and they were applied to evaluate the male distribution in the sampled Nannotrigona testaceicornis brood combs. MMC was very sensitive to slight deviations from a perfectly rounded clump; DMB detected any asymmetry in the location of these compact to fuzzy clusters. Six of the 82 brood combs of N. testaceicornis that we analyzed had more than nine males, distributed according to variations in spatial patterns, as indicated by the two indexes. PMID- 19551647 TI - Cuticular hydrocarbons in the stingless bee Schwarziana quadripunctata (Hymenoptera, Apidae, Meliponini): differences between colonies, castes and age. AB - Chemical communication is of fundamental importance to maintain the integration of insect colonies. In honey bees, cuticular lipids differ in their composition between queens, workers and drones. Little is known, however, about cuticular hydrocarbons in stingless bees. We investigated chemical differences in cuticular hydrocarbons between different colonies, castes and individuals of different ages in Schwarziana quadripunctata. The epicuticle of the bees was extracted using the non-polar solvent hexane, and was analyzed by means of a gas chromatograph coupled with a mass spectrometer. The identified compounds were alkanes, branched alkanes and alkenes with chains of 19 to 33 carbon atoms. Discriminant analyses showed clear differences between all the groups analyzed. There were significant differences between bees from different colonies, workers of different age and between workers and virgin queens. PMID- 19551648 TI - Comparative analysis of two sampling techniques for pollen gathered by Nannotrigona testaceicornis Lepeletier (Apidae, Meliponini). AB - Pollen counts from samples taken from storage pots throughout one year (from October to September) were adjusted by Tasei's volumetric correction coefficient for the determination of pollen sources exploited by two colonies of Nannotrigona testaceicornis in Sao Paulo, Brazil. The results obtained by this sampling technique for seven months (December to June) were compared with those from corbicula load samples taken within the same period. This species visited a large variety of plant species, but few of them were frequently used. As a rule, pollen sources that appeared at frequencies greater than 1% were found with both sampling methods and significant positive correlations (Spearman correlation coefficient) were found between their values. The pollen load sample data showed that N. testaceicornis gathered pollen throughout the external activity period. PMID- 19551649 TI - Trap-nesting bees (Hymenoptera: Apoidea) in forest fragments of the State of Sao Paulo, Brazil. AB - We studied the community ecology of trap-nesting bees in two forest fragments of the state of Sao Paulo, Brazil, during two years, utilizing bamboo canes and tubes made of black cardboard as trap nests. The traps were inspected once a month with an otoscope. One hundred and fifteen nests were obtained at Estacao Ecologica de Paulo de Faria, Paulo de Faria (EEPF). These included nine species belonging to five genera and two families. At Santa Cecilia Farm (SCF), 12 species belonging to seven genera and three families built 392 nests. Natural enemies reared from nests of both areas included Hymenoptera, Diptera and Coleoptera. Species richness was similar between the areas but the communities differed considerably in species composition. The higher diversity found at EEPF was due to more even distribution of the species. No difference was observed between the numbers of nests built in each year in each area. Although the species richness was lower in the cool/dry season of both years at SCF, and in the first year at EEPF, the nesting frequencies did not differ between seasons for both the overall community but for each of the most abundant species. No annual fluctuation in the frequencies of nesting was observed. As temperature and precipitation were not found to be significantly different between the two years of study in each area, we concluded that climatic stability resulted in population stability. PMID- 19551650 TI - Rearing Africanized honey bee (Apis mellifera L.) brood under laboratory conditions. AB - We developed a method for rearing larvae of Africanized bees under laboratory conditions to determine the amount of diet needed during larval development to obtain a worker bee. We started with larvae 18-24 h old, which were transferred to polyethylene cell cups and fed for five days. We found that the amount of diet needed for successful larval development was: 4, 15, 25, 50, and 70 microl during the first to fifth days, respectively. The survival rate to the adult stage was 88.6% when the larvae received the daily amount of diet divided into two feedings, and 80% when they received only one feeding per day. The adult weight obtained in the laboratory, when the larvae received the daily amount of diet in a single dose, did not differ from those that were developed under field conditions (our control). All adults that we obtained in laboratory appeared to be normal. This technique has the potential to facilitate studies on brood pathogens, resistance mechanisms to diseases and also might be useful to test the impacts of transgenic products on honey bee brood. PMID- 19551651 TI - [Hypertriglyceridemia]. PMID- 19551652 TI - [Intoxication with lithium]. AB - We report a case of a 75-year-old male patient who presented to the emergency room with arterial hypotension and impaired vigilance. The patient was on lithium therapy due to mood disorder. One month earlier medication with a betablocker, a loop-diuretic and an ACE-inhibitor had been started due to heart failure. Findings at admission included renal insufficiency, pneumonia and a slightly increased serum level of lithium. Three days later his Glasgow Coma Scale Score was 7, he showed gaze deviation, increased muscle tonus and cloni. The patient fully recovered after volume substitution and normalization of his renal function. Diagnosis of chronic intoxication with lithium was made due to the clinical picture and after exclusion of neurological pathologies. The pharmacokinetic characteristics of lithium is described and the risk factors leading to lithium intoxication and treatment of intoxication are discussed. PMID- 19551653 TI - [Comment on rule of thumb 3 in PRAXIS no. 7. "The patient who can climb on the cot with ease does not have appendicitis" ]. PMID- 19551654 TI - [Web based audiovisual patient information system--a pilot study of the preoperative patient information]. AB - A considerable part of the medical clinical daily routine incorporates the information of patients about medical diagnosis and especially about the scheduled operative intervention. The disease pattern, the course in the hospital before and after surgery and the operation itself will be discussed during these conversations. The patient has to be informed about all potential complications from a medico legal point of view. To optimise information delivery, auxiliary materials like charts, pictures, X-rays and models are used. Unfortunately, only a small percentage of the presented information is assimilated by the patient after strict verbal information. To inform patients better and earlier about medical details and internal processes before the operation, we developed a web based audiovisual patient information system with a combination of pictures, text, tone and video about surgical interventions. The patient satisfaction could be markedly improved by the application of this patient information system in the informed consent process. Furthermore, the use of the web-based multimedia information portal may leads to an essential time saving for the medical staff. PMID- 19551656 TI - [How to plan patient discharge from the hospital]. PMID- 19551655 TI - [Breathlessness: different causes and qualities of dyspnea]. AB - Dyspnea is the unpleasant awareness of breathing while healthy persons will hardly sense their own breath at rest. Dyspnea is comparable to pain because both caution the organism that will activate protective mechanisms to avoid further damage. Dyspnea results from central nervous processing of respiratory feed back signals. Feed back from respiratory muscles, the chest wall, the airways and the lung reach the brain while partial pressure of oxygen and carbon dioxide is continuously monitored and arterial blood gases are permanently stabilized. Sensation of breathlessness depends on its origin. Main qualities of dyspnea include air hunger during hypercapnia, laboured breathing during increased minute ventilation and chest tightness in asthma. During chronic dyspnea, respiratory control aims for a new breathing pattern that will ease breathlessness. Since respiratory control includes both autonomic regulation and cortical modulation of breathing, voluntarily guided respiration can be employed to influence other autonomic regulated systems and pain perception. PMID- 19551657 TI - [A 35-year old bricklayer with malperfusion of the left hand]. AB - A 35-year old man with coldness, paleness, numbness, and pain at Dig. IV of the left hand comes to the consultation. The Allen test is pathological, the angiography shows an occlusion at the length of 4 cm of the Arteria ulnaris beginning at the Hamulus ossis hamati. After therapy with prostaglandin E1, aspirin 100 mg for 10 days and a smoking stop, the symptom disappeared at the re consultation after 10 days. PMID- 19551663 TI - Effect of chalazion excision on refractive error and corneal topography. AB - PURPOSE: To evaluate refractive and corneal topographic changes following excision of chalazia. METHODS: This prospective noncomparative quasi-experimental clinical trial includes consecutive patients older than 7 years with chalazia of minimum duration of 1 month who underwent excision of the lesions by an internal or external approach. RESULTS: Overall, 253 lids from 228 eyes of 195 patients including 110 female subjects with mean age of 31-/+14 years (range 7-71) were studied. Mean duration of presenting symptoms was 4-/+2.8 months (range 1-24). Lesions were equally distributed in medial, central, and lateral areas of the eyelids. The chalazia were single in 172 (88.2%) and multiple in 23 (11.8%) patients. Mean change in best-corrected visual acuity, spherical equivalent refractive error, and difference of keratometry (corneal astigmatism) were 0.0004 /+0.007 logMAR (p=0.3), -0.06-/+0.6 D (p=0.1), and 0.34-/+0.35 (p<0.0001) after surgery, respectively. Corneal topographic analysis revealed significant changes in surface regularity index (0.13), surface asymmetry index (0.09), and potential visual acuity (0.06-/+0.1 logMAR) after the procedure (p<0.0001 for all comparisons). Single, central, and firm chalazia were compared with multiple, peripheral, and soft lesions; change in difference of keratometry and spherical equivalent were 0.75-/+0.42 D versus 0.22-/+0.15 D (p=0.001) and -0.35-/+0.19 versus -0.11-/+0.30 D (p=0.1), respectively, implying more induced astigmatism with the former type of lesions. CONCLUSIONS: Chalazion excision can decrease corneal astigmatism and irregularity, which is more prominent in single, firm, and central upper lid lesions. These findings may have implications in pediatric patients at risk for amblyopia. PMID- 19551664 TI - Extraocular muscle fixation to porous polyethylene orbital implants using 2-octyl cyanoacrylate. AB - PURPOSE: To evaluate the efficacy of bioadhesive in attaching the extraocular muscles to porous polyethylene spheres in rabbit enucleated cavities. METHODS: A prospective, experimental study was performed. Eight adult New Zealand white rabbits underwent right enucleation with insertion of an unwrapped 12 mm porous polyethylene sphere. The superior and inferior rectus muscles were attached to the implant with 2-octyl cyanoacrylate placed to the underside of the muscles and application of pressure for 5 seconds. Ninety days after surgery, the orbits were exenterated and four animals underwent histologic evaluation and the other four animals tensile strength testing of the muscle-sphere attachment. RESULTS: There were no exposures or infections of the implants and the muscles remained attached in all cases. Histologic examination showed fibrous tissue reaction at the muscle sphere attachment with minimal inflammation of the surrounding tissues; a pseudocapsule formation around the spheres and full vascularization of the implant was also observed. There was no evidence of residual glue on histologic examination. Tensile strength testing showed that rupture force was similar between bonded muscle and the porous polyethylene sphere. CONCLUSIONS: The 2 octyl-cyanoacrylate did not cause deleterious effects in orbital tissues and provided strong adhesion between muscles and spheres. Bioadhesive may be an option for attaching the extraocular muscles to orbital implants in anophthalmic cavity reconstruction. PMID- 19551665 TI - A new stereotest for strabismologists: the covered two pencil test. AB - PURPOSE: The two pencil test (TPT) checks the ability to perform a simple visual motor task. The test is only partially based on intact stereopsis, since monocular depth perception clues are involved. To overcome this shortcoming, a new test variant has been developed and tested. METHODS: By covering the tip of the examiner's rod, monocular cues should be drastically reduced. Twenty normal subjects performed the TPT and the covered two pencil test (CTPT) under monocular and binocular conditions. Each test condition was repeated 15 times. Four subjects were retested by three different examiners in order to determine the observer variation. Differences between the accuracy and observer variation of the two test variants under monocular and binocular condition were compared using the paired t test. RESULTS: The average difference between monocular and binocular accuracy was for the TPT 1.26 cm and for the CTPT 2.37 cm (p<0.0005). The average coefficients of variation of both test variants were similar, for the TPT 0.28 and for the CTPT 0.33 (p>0.1). CONCLUSIONS: This study demonstrates that covering the tip of the examiner's rod approximately doubles the differences between monocular and binocular accuracy. Therefore, covering seems to reduce monocular cues and increase the stereoscopic value of the TPT. PMID- 19551666 TI - Topographically guided two-step LASIK and standard LASIK in the correction of refractive errors after penetrating keratoplasty. AB - PURPOSE: To evaluate in a long-term period the effectiveness and safety of topographically guided two-step laser in situ keratomileusis (LASIK) and standard LASIK technique in the correction of refractive errors after successful penetrating keratoplasty (PKP) for keratoconus. METHODS: At least 2 years after PKP and 6 months after removal of all sutures, 15 eyes of 15 patients (Group 1; mean manifest refraction spherical equivalent (MRSE) -7.23 D -/+ 3.42 SD) were submitted to standard LASIK and 15 eyes of 15 patients (Group 2; mean MRSE -4.37 D -/+ 1.97 SD) to a topographically guided two-step LASIK procedure (first the flap and at least 2 weeks later the laser ablation). In all cases, a superior hinged corneal flap(160 microm/9.5 mm) was created. RESULTS: After a follow-up of 36 months, in Group 1 the mean uncorrected visual acuity (UCVA) was 0.51 logarithm of the minimum angle of resolution (logMAR) -/+ 0.41 SD and the mean best-corrected visual acuity (BCVA) was 0.03 logMAR -/+ 0.05 SD, with a mean MRSE of -1.57 D -/+ 2.65 SD. In Group 2, the mean UCVA was 0.28 logMAR -/+ 0.24 SD and the mean BCVA was 0.01 logMAR -/+ 0.03 SD, with a mean MRSE of -0.07 D -/+ 1.00 SD. In both groups, no complications were observed. CONCLUSIONS: After a long follow-up period, both topographically guided two-step LASIK and standard LASIK could be considered effective and safe tools in the correction of refractive errors after successful PKP for keratoconus. PMID- 19551667 TI - Aspheric profile versus wavefront-guided ablation photorefractive keratectomy for the correction of myopia using the Allegretto Eye Q. AB - PURPOSE: To compare the results of photorefractive keratectomy (PRK) with Q factor customized aspheric ablation and wavefront-guided customized ablation for correction of myopia compound with astigmatism. METHODS: Fifty-six patients were enrolled in this prospective controlled study. In each patient, one eye was randomly assigned for treatment with the Q-factor customized PRK (custom-Q study group) and the other eye treated with wavefront-guided customized PRK. Preoperative and 3-month postoperative manifest refraction, contrast sensitivity, visual acuity, aberrometry, and asphericity of the cornea were compared between the two groups. All eyes were treated with the Wavelight Eye-Q 400 Hz excimer laser in a single refractive surgery center by a single surgeon. RESULTS: Uncorrected visual acuity (UCVA) improved to 20/20 or more and 20/25 or more in 34 eyes (60.7%) and 56 eyes (100%), respectively, in the wavefront-guided ablation group and in 36 eyes (64.2%) and 54 eyes (96.4%) in the custom-Q ablation group. All eyes had UCVA of 20/40 or better. A total of 54 eyes (96.4%) in the wavefront-guided ablation group and 56 (100%) in the custom-Q ablation group had spherical equivalent (SE) within -/+0.5 D. One eye in each group (2%) lost >or=2 lines of best-corrected visual acuity (BCVA). High order root mean square (RMS) in the wavefront-guided group was 0.3630-/+0.13 mum preoperatively and 0.427-/+0.17 at 3 months (p=0.2). In the custom-Q ablation group it was 0.329 /+0.092 preoperatively and 0.4730-/+0.181 at 3 months after PRK (p=0.08). CONCLUSIONS: Regarding safety and refractive efficacy, custom-Q ablation profiles were clinically equivalent to wavefront-guided profiles in corrections of myopia up to -6.00 D and astigmatism up to 2.50 D. PMID- 19551669 TI - The efficacy of 5% povidone-iodine for 3 minutes prior to cataract surgery. AB - PURPOSE: To evaluate the efficacy of 3 minutes of 5% povidone-iodine for skin and conjunctival antisepsis prior to cataract surgery as recommended by the European Society of Cataract and Refractive Surgeons. METHODS: A prospective controlled study of 54 patients undergoing unilateral cataract surgery by phacoemulsification was undertaken. The eye undergoing surgery was prepared with 5% povidone-iodine for 3 minutes. The other eye served as control. Swabs were taken from both eyes prior to application of 5% povidone-iodine and 3 minutes after. Proportion of culture-positive swabs, mean number of species, and growth of isolates on culture were compared at different times and between the two groups. RESULTS: Coagulase-negative Staphylococcus (CoNS) was the main species isolated from culture-positive swabs. Use of 5% povidone-iodine for 3 minutes resulted in a statistically significant reduction in proportion of culture positive swabs (87% vs 30%, p<0.001), mean number of species (0.96-/+0.47 vs 0.30 /+0.46, p<0.001), and growth of CoNS (heavy: 30% vs 0%, moderate: 23% vs 6%, light: 47% vs 94%, p=0.004). No such changes were observed in the control group. CONCLUSIONS: Use of 5% povidone-iodine for 3 minutes prior to cataract surgery achieves a significant reduction in organisms such as CoNS from lid and conjunctival flora. PMID- 19551668 TI - Topical N-acetylcysteine reduces interleukin-1-alpha in tear fluid after laser subepithelial keratectomy. AB - PURPOSE: To evaluate the effect of topical N-acetylcysteine (NAC) on interleukin 1-alpha (IL-1alpha) levels in tear fluid after myopic laser subepithelial keratectomy (LASEK) and its possible role in modulating corneal wound healing. METHODS: Twenty-six eyes of 13 patients who underwent myopic LASEK were divided into 2 groups. Group 1 (n=10 eyes) was used as a control group. All patients received topical lomefloxacin and dexamethasone postoperatively. Additionally, patients in Group 2 received topical NAC for 1 month postoperatively. Tear fluid samples were collected with microcapillary tubes preoperatively, on the first and on the fifth postoperative day, and the release of IL-1alpha in tear fluid was calculated. Haze grading and confocal microscopic examination were performed at 1 and 3 months postoperatively. RESULTS: The mean IL-1-alpha release values were 0.285-/+0.159 pg/min in Group 1 and 0.235-/+0.142 pg/min in Group 2 preoperatively. In Group 1, the values were 0.243-/+0.155 pg/min on day 1 and 0.164-/+0.125 pg/min on day 5. In Group 2, the mean IL-1alpha release values were 0.220-/+0.200 pg/min on day 1 and 0.080-/+0.079 pg/min on day 5. The difference between the groups was significant only for day 5 (p<0.05). Mean corneal haze score and grey scale value in confocal microscopy were significantly higher (p<0.05) in Group 1 at 1 month. However, at 3 months there was no difference between groups (p>0.05). CONCLUSIONS: NAC seems to have an additive effect to steroids in suppressing IL-1alpha levels in tear fluid and may be clinically advantageous in modulating corneal wound healing during the early postoperative period after LASEK. PMID- 19551670 TI - Antioxidant status modifications by topical administration of dorzolamide in primary open-angle glaucoma. AB - PURPOSE: This study deals with the role of the topical administration of dorzolamide eyedrops on the oxidative/antioxidant status of aqueous humor in patients with primary open angle glaucoma (POAG). METHODS: A case-control study including 130 patients distributed into three groups was carried out: 1) patients with POAG without dorzolamide instillation administration (GG; n=34); 2) patients with POAG with dorzolamide (DG; n=36); and 3) subjects with cataracts (comparative group, CG; n=60). Oxidative activity was measured in the aqueous humor by malondialdehyde determination by the thiobarbituric acid reactive substances assay. Antioxidant status was assessed in the aqueous humor samples by measuring the superoxide dismutase activity and the total antioxidant status. RESULTS: Oxidative activity was significantly higher in both glaucoma groups than in the cataracts group (GG vs CG, p=3.68 E-34; DG vs CG, p=5.11 E-45) and was significantly higher in GG than in DG (p=0.0034). SOD activity was significantly higher in both glaucoma groups than in the cataracts group (GG vs CG, p=1.08 E 14; DG vs CG, p=3.70 E-22), and was significantly higher in GG than in DG (p=0.018). Finally, total antioxidant status was significantly decreased in both glaucoma groups compared with the cataracts group (GG vs CG, p=2.51 E-12; DG vs CG, p=5.06 E-05), and was more significantly decreased in GG than in DG (p=9.23 E 07). CONCLUSIONS: Topical administration of dorzolamide colirium diminishes oxidative stress in patients with glaucoma. PMID- 19551671 TI - Dry eye syndrome-related quality of life in glaucoma patients. AB - PURPOSE: To verify the presence of dry eye syndrome (DES) in treated patients with glaucoma and to analyze DES's impact on the patients' quality of life (QOL) versus the control group. METHODS: In this observational cross-sectional study, 61 patients were enrolled at a clinical practice. Patients were divided into three groups by number of glaucoma drops instilled per day (G1=1 drop/day, G2=2 drops/day, G3=3 drops/day). A control group of 20 subjects was also selected (G0). All subjects were submitted to a complete ocular examination (including tear function and ocular surface status) and completed the 25-item National Eye Institute Visual Function Questionnaire (NEI-VFQ), Glaucoma Symptom Scale (GSS) questionnaire, and Ocular Surface Disease Index (OSDI). DES was defined as presence of punctate keratitis and decreased break-up time. Statistical analysis was performed applying the Kruskal-Wallis analysis of variance and Mann-Whitney U tests (to compare median values between groups) as well as the chi2 and Fisher test (to verify significant differences). RESULTS: A total of 40% of G3 and 39% of G2 patients presented DES versus 11% of G1 and 5% of G0 (p=0.01). QOL was significantly influenced and altered (NEI-VFQ 25 total mean and GSS total mean and symptoms average: p=0.0085, p=0.006, and p=0.03, respectively). OSDI pointed out differences by group: 26% of G2 and 15% of G3 presented moderate OSDI and 15% of G3 and 8.7% of G2 severe OSDI (p>0.05). CONCLUSIONS: Patients with topically treated glaucoma present DES more often than a similar control group (p=0.01). The presence of DES negatively influences the patient's QOL. The patients with glaucoma's ocular surface status should be evaluated regularly to ensure the timely detection and treatment of pathologic signs on the ocular surface. PMID- 19551672 TI - Color Doppler imaging and ocular pulse amplitude in glaucomatous and healthy eyes. AB - PURPOSE: To determine factors influencing color Doppler imaging (CDI) measurements, to compare retrobulbar flow velocities between patients with glaucoma and healthy controls, and to describe the correlation between CDI and ocular pulse amplitude (OPA). METHODS: Patients with normal tension (n=28) or primary open angle glaucoma (n=19) and healthy controls (n=22) underwent CDI and OPA measurements. Intraocular pressure, corneal thickness, blood pressure, and heart rate were also measured. Spearman correlations were used to explore relations among these variables. A regression model for repeated measures was applied to compare between diagnostic groups the flow velocity indices in the retrobulbar vessels. RESULTS: Retrobulbar diastolic blood flow velocities correlated with diastolic blood pressure and perfusion pressure (range of Spearman rho [rho] coefficients=0.25-0.28; p=0.044-0.013 for the different vessels). Corneal thickness showed a positive correlation with systolic and diastolic flow velocities in the central retinal artery (rho=0.29 and 0.31; p=0.017 and 0.011 for peak systolic and end diastolic velocity, respectively). Systolic and diastolic blood flow velocities were reduced in the retrobulbar vessels of patients with normal tension glaucoma (P=0.0004) as well as primary open angle glaucoma (P=0.003) compared to healthy controls. A correlation was found between OPA and the resistive index in the retrobulbar vessels of the healthy controls (range rho=0.42-0.53; p=0.059-0.014). CONCLUSIONS: Retrobulbar blood flow velocities are reduced in patients with primary open angle glaucoma and normal tension glaucoma. Blood pressure and corneal thickness may influence CDI measurements. OPA correlates with the resistive index in CDI. PMID- 19551674 TI - A 12-week study evaluating the efficacy of bimatoprost 0.03% in patients with pseudoexfoliative and open-angle glaucoma. AB - PURPOSE: To evaluate the control of diurnal intraocular pressure (IOP) and the safety profile of bimatoprost in pseudoexfoliative glaucoma (PXG) compared to primary open angle glaucoma (POAG). METHODS: A prospective, observer-masked, nonrandomized study was performed. Seventy consecutive patients with either POAG (35 eyes) or PXG (35 eyes) drug-naive for glaucoma were assigned to receive bimatoprost 0.03% once daily for 12 weeks. Diurnal IOP was measured at baseline and after 12 weeks at three time points (8 AM, noon, and 4 PM). Main outcomes were diurnal IOP control and achievement of target IOP (CIGTS criteria). Mean diurnal IOP, hour-by-hour IOP measurements, and safety, including serious adverse events, were also evaluated. RESULTS: A significant IOP reduction from baseline was found in both groups (p<0.001). Mean and hour-by-hour IOP differences between groups were not statistically significant (NS). The observed IOP values and percentages of IOP reduction were 17.0 mmHg (31.5%) and 16.4 mmHg (31.9%) in PXG and POAG eyes, respectively; the differences were not statistically significant. Six eyes (1 POAG and 5 PXG, respectively) responded with a <20% IOP reduction (NS). Twenty-seven POAG (77.1%) and 23 PXG (65.7%) eyes achieved target IOP. Consequently,20 eyes (8 POAG and 12 PXG, respectively) were classified as unable to achieve the IOP target values (NS). CONCLUSIONS: Bimatoprost was effective and safe in lowering IOP both in open angle and pseudoexfoliative glaucoma, achieving target pressure in most patients. However, long-term efficacy in PXG must be evaluated. PMID- 19551673 TI - Ophthalmologist attitudes regarding fixed combination treatment for glaucoma in the European Union. AB - PURPOSE: To survey ophthalmologists in the European Union to determine their fixed combination (FC) preferences. METHODS: A multiple-choice survey was sent to randomly chosen ophthalmologists throughout the European Union and bordering countries by electronic mail delivery. Ophthalmologists were resent the survey on two more occasions if no response was obtained. RESULTS: In total, 50 surveys were received from 530 distributed (9.4% response) from 16 countries. More ophthalmologists prescribed an FC as second (80%) or third choice (64%) therapy than first choice (30%, p=0.0036). As first (p<0.0001) and third choice (p=0.011), the dorzolamide/timolol FC was most commonly prescribed, while as second choice the latanoprost/timolol FC (p<0.0001) was most popular. Overall, 98% (49/50) of doctors believed FC therapy improved patient care most often by better compliance (n=49) and quality of life(n=48, p<0.0001). Most ophthalmologists believed that there was a difference in efficacy between the FC products (32/50, 64%) with the most effective being the bimatoprost/timolol FC (n=12, 24%, p=0.029). However, fewer ophthalmologists perceived safety differences between the products (34/50, 68%). For prostaglandins, dosing time was suggested in the morning by 18 (36%) ophthalmologists and in the evening by 24 (48%) (p=0.35). CONCLUSIONS: FCs in the European Union are a potentially popular method to reduce intraocular pressure, being prescribed most commonly as second or third choice therapy. The perceived advantages to FC therapy are greater adherence to the medicine and improved patient quality of life. PMID- 19551675 TI - Ultrasound biomicroscopy in nonperforating filtering surgery with intraoperative 5-fluorouracil and no implant: long-term results. AB - PURPOSE: To evaluate the long-term morphologic changes in the anterior segment structures after nonpenetrating filtering surgery (NPFS) supplemented with 5 fluorouracil (5-FU) and no scleral implant using ultrasound biomicroscopy (UBM). METHODS: Thirteen eyes of 13 consecutive patients who underwent NPFS with intraoperative 5-FU under the conjunctiva and the scleral flap and no implant were evaluated in an observational nonrandomized, consecutive case series study conducted 2 years postoperatively. Patients were assessed for the presence of a subconjunctival filtering bleb, the volume of an intrascleral cavity, and a suprachoroidal hypoechoic area. The intraocular pressure(IOP) was measured preoperatively and postoperatively at the time of UBM. RESULTS: The IOP decreased significantly (p=0.01) from 24-/+7.6 mmHg to 13.7-/+4.1 mmHg. In most patients (69.2%), the postoperative IOP decreased at least 30% from the preoperative value without medications and in 84.6% with medication. UBM showed a subconjunctival empty space in 92.3% of eyes. In 84.6% of patients, an intrascleral cavity was seen, the mean volume of which was 1.68 mm(3) (range, 0-4.07). We found a negative correlation between the height, width, and volume of the intrascleral lake and the IOP. In 92.3% of eyes, a hypoechoic area in the suprachoroidal space also was seen. CONCLUSIONS: UBM showed a filtering intrascleral cavity, subconjunctival filtering bleb, and a suprachoroidal space after NPFS supplemented with 5-FU under the conjunctiva and the scleral flap without an implant. PMID- 19551676 TI - Intravitreal bevacizumab as an adjunct treatment for neovascular glaucoma. AB - PURPOSE: To assess the effect of adjunctive intravitreal bevacizumab treatment on neovascular glaucoma (NVG). METHODS: The medical records of all consecutive patients with NVG treated with intravitreal bevacizumab at our center from May 2006 to February 2008 were reviewed. The data collected included background features, findings on full ophthalmologic examination (including visual acuity, gonioscopy, and intraocular pressure), glaucoma medications prescribed, and additional procedures for glaucoma performed before and after bevacizumab injection.The interval between the diagnosis of NVG and intravitreal bevacizumab treatment was calculated. RESULTS: Eighteen patients (6 male, 12 female; mean age 63-/+13.2 years) met the study criteria. Causes of NVG were proliferative diabetic retinopathy (n=14), central retinal vein occlusion (n=2), occlusive vasculitis (n=1), and panuveitis (n=1). The mean duration of followup was 52 ( /+12) weeks. Mean intraocular pressure decreased from 32.3 (-/+4.99) to 18 ( /+6.1) mmHg (p<0.0001) and mean number of glaucoma medications decreased from 3.16 (-/+1.2) to 2.55 (-/+1.46) (p=0.1938). An interval of less than 6 months between the start of bevacizumab treatment and diagnosis was associated with better final visual acuity than delayed treatment (0.82-/+0.4 logMAR vs 1.88 /+1.1 logMAR, p=0.002) and a better regression of iris neovascularization (22% vs 89%; p=0.015). CONCLUSIONS: Intravitreal bevacizumab is beneficial for the treatment of anterior segment neovascularization and NVG when used as an adjunct, making the administration of additional treatment for the underlying cause possible. Bevacizumab should be instituted promptly after diagnosis, before irreversible anatomic and functional damage occurs. PMID- 19551677 TI - Intravitreal bevacizumab for treatment of chronic central serous chorioretinopathy. AB - PURPOSE: To evaluate the short-term safety and efficacy of intravitreal bevacizumab for the treatment of intraretinal or subretinal fluid accumulation secondary to chronic central serous chorioretinopathy (CSC). METHODS: Twelve patients were treated with intravitreal injections of 2.5 mg bevacizumab at 6- to 8-week intervals until intraretinal or subretinal fluid resolved. Observation procedures were Early Treatment Diabetic Retinopathy Study best-corrected visual acuity (BCVA), ophthalmic examination, and optical coherence tomography (OCT), performed at 6- to 8-week intervals. Fluorescein angiography was performed at baseline visit and thereafter depending on clinical and OCT findings. Multivariate analysis of variance with repeated measures was used to calculate a statistical significance of change in BCVA and mean central retinal thickness, which were the main outcome measures. SAS statistical software was used for analyses. RESULTS: Patients received 2-/+1 intravitreal injections of bevacizumab on average during a follow-up of 24-/+14 weeks. Mean BCVA increased by 2-/+2 lines; the change in BCVA (logMAR) was significant (p<0.02). Mean central retinal thickness decreased significantly over follow-up (p<0.05), with 6 patients (50%) showing complete resolution of subretinal fluid. CONCLUSIONS: Anatomic and functional improvement following intravitreal bevacizumab injections suggest that vascular endothelial growth factor (VEGF) may be involved in fluid leakage in patients with chronic CSC. The results suggest a possible role for anti-VEGF agents in the treatment of chronic CSC. Further evaluation of intravitreal bevacizumab for chronic CSC in controlled randomized studies is warranted. PMID- 19551678 TI - Can an intraoperative bevacizumab injection prevent recurrent postvitrectomy diabetic vitreous hemorrhage? AB - PURPOSE: To evaluate the recurrence of vitreous hemorrhage (VH) in patients treated with intravitreal bevacizumab (IVB) injection (2.5 mg/0.1 mL) intraoperatively at the end of vitrectomy for treatment of diabetic nonclearing VH. METHODS: A prospective pilot study of 30 eyes of 28 consecutive diabetic patients who underwent pars plana vitrectomy and IVB injection intraoperatively at the end of vitrectomy was performed. The amount of VH was graded with slit lamp biomicroscopy by three masked retinal specialists from grade 0 to grade 3. Main outcome measures were rate of recurrence of the VH, improvement in visual acuity, incidence of cataract formation, and postoperative complications through a follow-up of 6 months. RESULTS: The percentage of severe recurrent VH with no fundus details (grade 3) was 7%, 13%, 27%, and 30%, respectively, at 7 days and 1 , 3-, and 6-month follow-up. At 6-month followup, the best-corrected visual acuity improved from 1.00 to 0.4 logMAR (p=0.01) in 21 out of 30 eyes (70%). Nine out 20 (40%) phakic patients developed cataract during the follow-up period, and 7 (31%) of them underwent cataract surgery. CONCLUSIONS: The study suggests that intravitreal bevacizumab injection cannot prevent rebleeding in eyes undergoing pars plana vitrectomy for treatment of diabetic vitreous hemorrhage. PMID- 19551679 TI - Intravitreal bevacizumab in refractory uveitic macular edema: one-year follow-up. AB - PURPOSE: Uveitis is a major cause of ocular morbidity in developed countries. It has been demonstrated that macular edema is a significant cause of decreased visual acuity and macular edema in these patients. In this article, we evaluate the long-term outcome of intravitreal bevacizumab in the treatment of refractory uveitic macular edema. METHODS: In this retrospective, noncomparative, interventional case series, uveitic patients with macular edema who were refractory to conventional therapy and who were treated with intravitreal bevacizumab were identified and assessed. Best-corrected visual acuity and optical coherence tomography central macular thickness measurements were collected and analyzed with correlative statistical analysis, including the use of Student paired t-test, Kaplan-Meier, and linear regression analysis. RESULTS: Twenty-nine eyes of 27 patients with diverse uveitic etiologies were analyzed and followed up at 1 year. Thirteen patients received a single intravitreal bevacizumab injection. Six patients required a second intravitreal bevacizumab injection, while 10 patients received combination therapy of intravitreal bevacizumab and triamcinolone acetonide. Baseline mean logMAR visual acuity was 0.59. At 1 year, the mean logMAR visual acuity was -0.42-/+ 0.36 (p=0.0045). Baseline mean central macular thickness was 383.66 microm. At 1 year, the mean thickness was 294.32-/+110.87 (p=0.0007). CONCLUSIONS: Intravitreal bevacizumab is a useful and therapeutically beneficial agent in the treatment of refractory uveitic macular edema. Some patients will require adjunctive intravitreal bevacizumab injections or the use of combination therapy with intravitreal triamcinolone acetonide. PMID- 19551680 TI - Efficacy of intravitreal triamcinolone after or concomitant with laser photocoagulation in nonproliferative diabetic retinopathy with macular edema. AB - PURPOSE: To investigate the clinical effects and outcomes of intravitreal injection of 4 mg of triamcinolone acetonide (IVTA) after or concomitant with macular laser photocoagulation (MP) for clinically significant macular edema (CSME). METHODS: Forty-nine eyes of 49 patients with nonproliferative diabetic retinopathy and CSME were randomized into three groups. The eyes in the laser group (n=17), group 1, were subjected to MP 3 weeks after IVTA; the eyes in the IVTA group (n=13), group 2, were subjected to MP, concomitant with IVTA; the eyes in the control group (n=19), group 3, underwent only IVTA application. Visual acuity (VA), fundus fluorescein angiography, and photography were performed in each group. RESULTS: In the first group, the mean VA improved from 0.17-/+0.09 at baseline to 0.28-/+0.15 (p=0.114) and in the second group, deteriorated from 0.19 /+0.08 at baseline to 0.14-/+0.08 at the sixth month (p=0.141), respectively. In Group 3, the mean VA improved from 0.16-/+0.08 at baseline to 0.28-/+0.18 (p=0.118) at the end of the follow-up. When VA was compared between the control and study groups, significant difference was detected at the sixth month (p=0.038). CONCLUSIONS: MP after IVTA improved VA, rather than MP concomitant with IVTA, and only IVTA application for CSME. MP after IVTA may reduce the recurrence of CSME and needs further investigations in a longer period. PMID- 19551681 TI - HLA-DQB1 subtypes predict diabetic retinopathy in patients with type I diabetes mellitus. AB - PURPOSE: To investigate if diabetic patients without diabetic retinopathy despite long disease duration have different human leukocyte antigen (HLA) status vs those with an early onset of retinopathy. METHODS: Retrospective, nonrandomized, masked comparative study. Type 1 diabetic patients with a disease onset before age 30 were recruited to the study. The study population consisted of two groups of diabetic patients: those with normal retinopathy course (retinopathy developed during the first 20 years of diabetes onset) (23 patients) and those with postponed retinopathy (no obvious retinopathy in spite of passing 20 years of diabetes) (19 patients). These groups were matched with regard to level of glycemic control, blood pressure, and lipid profile. A group of 23 healthy patients served as controls. HLA-DQB1 typing of blood samples was done using a polymerase chain reaction with sequence-specific primer (PCR-SSP) method. RESULTS: HLA-DQB1*0201/HLA-DQB1*0501 and HLA-DQB1*0201/HLA-DQB1*0504 haplotypes were more common among type 1 diabetic patients with normal retinopathy course than those with postponed retinopathy (26.1% vs 0.0%; p=0.019). HLA-DQB1*0301 and HLA-DQB1*0304 were less common among those diabetic patients with normal retinopathy course than those with a postponed retinopathy (63.2% vs 34.8%; p=0.067). CONCLUSIONS: Some haplotypes seem to predispose diabetic patients to diabetic retinopathy. HLA typing may be beneficial for predicting the prognosis of diabetic retinopathy in younger diabetic patients. PMID- 19551682 TI - Long-term choroidal vascular changes after iodine brachytherapy versus transpupillary thermotherapy for choroidal melanoma. AB - PURPOSE: To compare long-term choroidal vascular changes after iodine-125 brachytherapy (IBT) versus transpupillary thermotherapy (TTT) used as primary treatment of small choroidal melanoma. METHODS: Ninety-five small choroidal melanomas were randomized: 49 eyes with TTT and 46 eyes with IBT alone. Fluorescein and indocyanine green angiography (ICGA) were performed at 3-month intervals during the first year, and every 6 months thereafter. RESULTS: Mean follow-up was 56.2 months (range, 24-118 months; SD, 22.6). Tumor regressed in 45 (92%) TTT-treated vs 45 (98%) IBT-treated eyes (p=0.397). Four TTT-treated and one IBT-treated tumor recurred. Occlusion of choriocapillaris was present in all TTT and IBT cases. Closure of medium and large choroidal vessels was observed in 17 (35%) TTT-treated vs 44 (96%) IBT-treated eyes (p<0.001). Choroidal vascular remodeling was detected in 20 (41%) TTT-treated and 16 (35%) IBT-treated eyes (p=0.693). Retinochoroidal anastomosis was present in 4 of the 37 (11%) TTT treated eyes with patency of medium and large choroidal vessels, but never observed in the IBT-treated eyes, and was associated with tumor recurrence. Among IBT-treated eyes, segments of choroidal vascular wall ICG staining and choroidal aneurysmal changes were detected in 30 (65%) and 7 (15%), respectively. These changes were never detected in TTT-treated cases (p<0.0001 and p=0.015, respectively). CONCLUSIONS: The pattern of tumor choroidal vascular changes following IBT and TTT differs. TTT is less effective in closing all tumor vasculature. The role of long-term choroidal vascular remodeling observed after these two treatments needs longer follow-up. PMID- 19551683 TI - Proton beam radiotherapy of uveal melanoma: Italian patients treated in Nice, France. AB - PURPOSE: To evaluate the results of 15 years of experience with proton beam radiotherapy in the treatment of intraocular melanoma, and to determine univariate and multivariate risk factors for local failure, eye retention, and survival. METHODS: A total of 368 cases of intraocular melanoma were treated with proton beam radiotherapy at Centre Lacassagne Cyclotron Biomedical of Nice, France, between 1991 and 2006. Actuarial methods were used to evaluate rate of local tumor control, eye retention, and survival after proton beam radiotherapy. Cox regression models were extracted to evaluate univariate risk factors, while regularized least squares algorithm was used to have a multivariate classification model to better discriminate risk factors. RESULTS: Tumor relapse occurred in 8.4% of the eyes, with a median recurrence time of 46 months. Enucleation was performed on 11.7% of the eyes after a median time of 49 months following proton beam; out of these, 29 eyes were enucleated due to relapse and 16 due to other causes. The univariate regression analysis identified tumor height and diameter as primary risk factors for enucleation. Regularized least squares analysis demonstrated the higher effectiveness of a multivariate model of five risk factors (macula distance, optic disc distance, tumor height, maximum diameter, and age) in discriminating relapsed vs nonrelapsed patients. CONCLUSIONS: This data set, which is the largest in Italy with relatively long term follow-up, demonstrates that a high rate of tumor control, survival, and eye retention were achieved after proton beam irradiation, as in other series. PMID- 19551684 TI - Aqueous concentration of tranexamic acid after 5% and 10% eyedrop administration. AB - PURPOSE: To determine whether topical tranexamic acid can provide acceptable therapeutic concentration in the aqueous humor. METHODS: A total of 47 patients with cataract as the sole eye disease who have been scheduled for cataract operation received one drop of tranexamic acid 5% solution at various time intervals prior to operation (from 15 minutes to 9 hours). Two patients received 10% solution of the drug. Paracentesis of the anterior chamber was done and the aqueous humor was aspirated in the operation room. The aqueous samples were analyzed using high-performance liquid chromatography method with the limit of quantification around 0.1 microg/mL. Systemic and ocular side effects were evaluated. RESULTS: Aqueous concentrations of tranexamic acid was higher than 1.5 microg/mL up to 160 minutes after administration of a single drop of 5% solution of the drug and then declined to an average concentration of 1 microg/mL at 300 minutes which persisted up to 9 hours after administration.In two patients for whom a single drop of 10% tranexamic acid solution was used, aqueous concentrations of 2.72 and 2.90 microg/mL were detected 60 minutes after administration. None of the patients experienced ocular or systemic side effects. CONCLUSIONS: Topical administration of tranexamic acid is effective in yielding therapeutic intraocular concentrations of drug without any ocular or systemic toxicity. PMID- 19551685 TI - KIF21A variant R954W in familial or sporadic cases of CFEOM1. AB - PURPOSE: To demonstrate the clinical characteristics and determine mutations in the KIF21A gene, encoding a kinesin motor protein in patients with congenital fibrosis of the extraocular muscles (CFEOM) type 1. METHODS: Patients of five families with congenital fibrosis syndrome and two simplex patients with CFEOM underwent ophthalmologic examination and mutation analysis in the KIF21A gene. RESULTS: Clinical examination and passive motility testing prior to surgery met criteria for CFEOM. All patients had congenital restrictive ophthalmoplegia primarily affecting muscles innervated by the oculomotor nerve. Complete mutation screening in the KIF21A gene revealed the presence of the known and most common recurrent variant R954W in three families and in two simplex cases. Two families demonstrated linkage to chromosome 16. CONCLUSIONS: The patients included in the study had marked restriction of movement bilaterally with nearly complete loss of vertical ocular motility, graded reduction of horizontal motility, ptosis, and compensatory chin elevation. The phenotype was variable in patients carrying the same mutation. In one family, all patients were diagnosed with mental retardation, indicating that this syndrome might not only affect the development of cranial nerves, but can also be responsible for general neurologic dysfunction. The screening data suggest frequent and exclusive appearance of the R454W variant in sporadic and familial cases of CFEOM1 in Germany. PMID- 19551686 TI - Recently recognized cases of ophthalmofilariasis in Hungary. AB - PURPOSE: To report four cases of zoonotic ophthalmodirofilariasis infection caused by Dirofilaria repens in Hungary. METHODS: Four cases of ophthalmofilariasis have been treated at our department during the last 14 months. A subconjunctival moving worm was observed by slit lamp biomicroscopy in two cases. In one of these a living filaria was surgically removed, but the other disappeared. Red eye and migrating edema were the presenting signs in two cases. A biopsy taken from the subcutaneous masses disclosed D repens. RESULTS: Histopathologic or parasitologic examination identified a female D repens in every case. Laboratory alterations were not found. Symptoms subsided after treatment. CONCLUSION: The clinical presentation of filariasis is not always straightforward, and a high index of suspicion is necessary in cases presenting with orbital or periorbital inflammation. During the past 10 years the identification of locally acquired infections by D repens has increased in Hungary. PMID- 19551687 TI - Acute presentation of nasal-type natural killer/T-cell lymphoma of the orbit. AB - PURPOSE: An unusual case of nasal-type natural killer/T-cell lymphoma (NKTL) of the orbit is reported. METHODS: The clinical history, computed tomography, magnetic resonance imaging, and biopsy specimen of a 29-year-old man with a right orbital lymphoma were evaluated. RESULTS: The patient initially presented with conjunctival injection and had flu-like symptoms before developing right proptosis and reduced vision; imaging showed a diffuse infiltrative process throughout the orbit. Orbital biopsy revealed angiodestruction with prominent necrosis, and angiocentric lymphoma growth and lymphoma cells were positively stained for CD3, CD20, CD45RO, CD56, cytotoxic molecules (granzyme B and T-cell intracellular antigen-1), and Epstein-Barr virus. CONCLUSIONS: NKTL is rare and may present acutely; the imaging findings presented serve to highlight the radiologic features of the disease. PMID- 19551688 TI - Palpebro-orbital myiasis in a patient with basal cell carcinoma. AB - PURPOSE: We report a patient with basal cell carcinoma presenting with severe myiasis in a large ulcer involving the upper and lower eyelid. METHODS: Myiasis is an infestation of vertebrate animals by larvae of certain fly species. About 70 larvae were removed manually. A biopsy of the tissue underneath demonstrated a basal cell carcinoma. The myiasis was produced by the fly Lucilia sericata, currently used for treating chronic nonhealing ulcers. Because of the stage of the carcinoma, an orbital exenteration was carried out. RESULTS: The patient died 2 days later because of cardiopulmonary failure not related to the myiasis. CONCLUSIONS: We present a case of a severe orbital myiasis focusing on its management and life-threatening nature. PMID- 19551689 TI - Iris color: validation of a new classification and distribution in a Spanish population-based sample. AB - PURPOSE: To design and validate a standard, simple, and reliable iris color classification and to study its distribution in a Spanish population. Iris color has a geographic distribution and has been correlated with different ocular diseases. However, there is no standard and validated iris color classification allowing comparison among different studies. METHODS: Classification was made in three grades (blue-gray, hazel-green, brown) and was validated by 3 independent readers. Initially, a preliminary study was made in 50 iris photographs to detect technical hitches. Afterwards, based on this procedure, 221 iris photographs were graded. RESULTS: Measures of interobserver reliability were 0.786 by kappa index with an agreement of 89.6%.Iris color distribution in the Spanish cohort was blue grey 16.29%, hazel-green 55.2%, and brown 28.5%. CONCLUSIONS: This classification is simple, reliable, and easy to use in clinical research and by ophthalmologists or generalists in practice. The Spanish cohort from this study shows a different iris color distribution from those previously published in other countries. PMID- 19551690 TI - Leber miliary aneurysms and multiple sclerosis. AB - PURPOSE: To describe a patient with Leber miliary aneurysms who fulfilled the criteria for clinically definite multiple sclerosis (MS) in remission. METHODS: Case report. RESULTS: We performed a complete ophthalmologic examination in a 45 year-old woman who was referred to our department for retinal vascular abnormalities in her left eye. Twenty-eight years previously,the patient had been diagnosed with MS. Following a complete ophthalmologic examination,the patient was diagnosed with Leber miliary aneurysms and coincident MS in remission. To our knowledge, there are no previous reports describing association between Leber miliary aneurysms and MS. CONCLUSIONS: Leber miliary aneurysms have been found in association with multiple ocular and systemic syndromes. In this report, we suggest another possible systemic association (MS) with Leber miliary aneurysms. PMID- 19551691 TI - Cardiorespiratory depression and hyperglycemia after unintentional ingestion of brimonidine in a neonate. AB - PURPOSE: Brimonidine is a third-generation selective alpha-2 adrenergic agonist that lowers intraocular pressure by decreasing aqueous humor production and increasing uveoscleral flow. Its safety profile for children <2 years of age remains unknown. METHODS: We describe a case of ingestion of a single drop of brimonidine 0.2% in a newborn, resulting in sedation, cardiorespiratory depression, and hyperglycemia within minutes. CONCLUSIONS: This report adds to the existing evidence that brimonidine can have serious adverse side effects and should therefore be used with extreme caution in infants <2 years of age. Additionally,safety procedures like double-checking should also include vitamins or oral supplements to improve medication safety in the neonatal intensive care unit. PMID- 19551693 TI - Calcium isotope analysis by mass spectrometry. AB - The variations in the isotopic composition of calcium caused by fractionation in heterogeneous systems and by nuclear reactions can provide insight into numerous biological, geological, and cosmic processes, and therefore isotopic analysis finds a wide spectrum of applications in cosmo- and geochemistry, paleoclimatic, nutritional, and biomedical studies. The measurement of calcium isotopic abundances in natural samples has challenged the analysts for more than three decades. Practically all Ca isotopes suffer from significant isobaric interferences, whereas low-abundant isotopes can be particularly affected by neighboring major isotopes. The extent of natural variations of stable isotopes appears to be relatively limited, and highly precise techniques are required to resolve isotopic effects. Isotope fractionation during sample preparation and measurements and instrumental mass bias can significantly exceed small isotope abundance variations in samples, which have to be investigated. Not surprisingly, a TIMS procedure developed by Russell et al. (Russell et al., 1978. Geochim Cosmochim Acta 42: 1075-1090) for Ca isotope measurements was considered as revolutionary for isotopic measurements in general, and that approach is used nowadays (with small modifications) for practically all isotopic systems and with different mass spectrometric techniques. Nevertheless, despite several decades of calcium research and corresponding development of mass spectrometers, the available precision and accuracy is still not always sufficient to achieve the challenging goals. The present article discusses figures of merits of presently used analytical methods and instrumentation, and attempts to critically assess their limitations. In Sections 2 and 3, mass spectrometric methods applied to precise stable isotope analysis and to the determination of (41)Ca are described. Section 4 contains a short summary of selected applications, and includes tracer experiments and the potential use of biological isotope fractionation in medical studies, paleoclimatic and paleoceanographic, and other terrestrial as well as extraterrestrial investigations. PMID- 19551695 TI - Electrospray: from ions in solution to ions in the gas phase, what we know now. AB - There is an advantage for users of electrospray and nanospray mass spectrometry to have an understanding of the processes involved in the conversion of the ions present in the solution to ions in the gas phase. The following processes are considered: Creation of charge droplets at the capillary tip; Electrical potentials required and possibility of gas discharges; Evolution of charged droplets, due to solvent evaporation and Coulomb explosions, to very small droplets that are the precursors of the gas phase ions; Production of gas phase ions from these droplets via the Ion Evaporation and Charge residue models; Analytical uses of ESIMS of small ions, qualitative and quantitative analysis; Effects of the ESI mechanism on the analysis of proteins and protein complexes; Determination of stability constants of protein complexes; Role of additives such as ammonium acetate on the observed mass spectra. PMID- 19551694 TI - Unilateral and bilateral upper extremity weight-bearing effect on upper extremity impairment and functional performance after brain injury. AB - The purpose of the study was to investigate the effect of upper extremity (UE) weight bearing on UE impairment functional performance of persons with acquired brain injury (BI). A quasi-experimental design was used to examine a convenience sample of 99 persons with acquired BI and 22 without BI (WBI) living in a community re-entry centre. A computerized force-sensing array pressure map system was used to determine the UE pressure during unilateral and bilateral conditions. Differences between groups were examined using t-tests. Correlations were computed between UE weight bearing and hand function, and functional performance as measured by the Fugl-Meyer scale and functional independence measure (FIM) scale. The group of people with BI exerted significantly lower UE weight bearing during unilateral conditions as compared with persons WBI [left: t (119) = 2.34, p = 0.021; right: t (119) = 4.79, p = 0.043). UE weight-bearing measures correlated strongly with FIM motor scores with bilateral UE conditions yielded the highest significant correlation (bilateral left r = 0.487, p < 0.001; bilateral right r = 0.469, p < 0.01). The results indicated that UE weight bearing pressure differs in unilateral and bilateral conditions, between persons with and WBI and between persons with stroke and traumatic brain injury. These findings may have implications for occupational therapists that use unilateral versus bilateral motor training for rehabilitation. There is a need to replicate the study design with a randomized and stratified sample of persons with BI. PMID- 19551697 TI - Montelukast and psychiatric disorders in children. AB - PURPOSE: A signal has been raised concerning montelukast and adverse drug reactions (ADRs) in children. The purpose of the present study was to evaluate psychiatric ADRs during treatment with montelukast in children. METHODS: We analyzed all reports of psychiatric disorders during treatment with montelukast in children (<18 years) in the Swedish ADR database SWEDIS (1998-2007). The Bayesian Confidence Propagation Neural Network (BCPNN) method was used to screen for disproportional reporting rates of these reactions. An information component (IC) with a positive 95% lower confidence limit indicates a statistically significant disproportionality between the expected and the reported rate for a drug and an ADR. RESULTS: A total of 48 reports of psychiatric disorders in children during treatment with montelukast were found in SWEDIS. Reports appeared every year after registration. Psychiatric disorders reported more than once included nightmares (n = 15), unspecified anxiety (n = 11), aggressiveness (n = 11), sleep disorders (n = 10), insomnia (n = 3), irritability (n = 3), hallucination (n = 3), hyperactivity (n = 3), and personality disorder (n = 2). In 23 reports (48%), the child experiencing psychiatric ADRs was < or = 3 years old. Time from exposure to ADR was indicated in 35 reports. In 28 of these (80%), the time from exposure to ADR was less than 1 week. A statistical signal for psychiatric disorders appeared in the fourth quarter of 1998 (three reports, IC value: 2.34, 95% lower confidence limit: 0.62). CONCLUSIONS: Psychiatric ADRs can occur during montelukast treatment in children, indicating that attention to this is essential. Further studies are needed to establish the magnitude of the problem. PMID- 19551696 TI - Anterior cingulate cortical volumes and treatment remission of geriatric depression. AB - BACKGROUND: Structural abnormalities of the anterior cingulate cortex (ACC) may interfere with the interaction of cortical and limbic networks involved in emotional regulation and contribute to chronic depressive syndromes in the elderly. This study examined the relationship of regional anterior cingulate cortical volumes with treatment remission of elderly depressed patients. We hypothesized that patients who failed to remit during a 12-week controlled treatment trial of escitalopram would exhibit smaller anterior cingulate gray matter volumes than patients who remitted. METHODS: The participants were 41 non demented individuals with non-psychotic major depression. After a 2-week single blind placebo period, subjects who still had a Hamilton Depression Rating Scale (HDRS) of 18 or greater received escitalopram 10 mg daily for 12 weeks. Remission was defined as a HDRS score of 7 or below for at least 2 consecutive weeks. The patient sample consisted of 22 depressed patients who achieved remission during the study and 19 depressed patients who remained symptomatic. High-resolution magnetization-prepared rapidly acquired gradient echo (MPRAGE) sequences were acquired on a 1.5 T scanner and regional ACC volumes were manually outlined (dorsal, rostral, anterior subgenual, and posterior subgenual). RESULTS: Repeated measure analyses revealed that patients who failed to remit following escitalopram treatment had smaller dorsal and rostral anterior cingulate gray matter volumes than patients who remitted, whereas subgenual cortical volumes did not differ between the groups. CONCLUSIONS: Structural abnormalities of the dorsal and rostral anterior cingulate may perpetuate late-life depression. PMID- 19551698 TI - Antiretroviral induced adverse drug reactions in Iranian human immunodeficiency virus positive patients. AB - PURPOSE: To our knowledge, there is no report regarding antiretroviral induced adverse drug reactions (ADRs) in Iranian patients who infected with human immunodeficiency virus (HIV). We have evaluated the frequency of antiretroviral therapy (ART) induced ADRs in this population and assessed some risk factors of these reactions. METHODS: This is a prospective cohort study that was performed in Iranian Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome Research Center during years 2005-2007. Adult patients who infected with HIV and newly started on ART were included in this study and followed laboratory and clinically for the development of any ADRs for at least 6 months. RESULTS: During this study 87.6% of patients showed at least one ADR. Prevalence of ADRs based on affected organ was: gastrointestinal (GI) (63.7%), hematological (32.6%), neurological (30%), cutaneous (22%), musculoskeletal (21.3%), hepatic (20%), metabolic (18.6%), and renal (2.6%) adverse effects. CONCLUSIONS: The most important results of this study were finding of hematological ADRs as the most common cause of ART interruption, supporting the hypersensitivity nature of antiretroviral induced hepatotoxicity especially cholestasis and new finding of the significant positive correlation between hepatitis C virus co-infection and the occurrence of ART induced skin reactions. PMID- 19551699 TI - Validity of The Health Improvement Network (THIN) for epidemiologic studies of hepatitis C virus infection. AB - PURPOSE: Before using computerized databases to study hepatitis C virus (HCV) epidemiology, the validity of the diagnosis must be assessed. We determined the accuracy of HCV diagnostic codes within The Health Improvement Network (THIN), an electronic database containing medical record data from general medical practices in the United Kingdom. METHODS: Patients with initial diagnostic codes for HCV infection and nonspecific viral hepatitis between 2000 and 2007 in the THIN database were identified. Questionnaires were mailed to general practitioners caring for a random sample of 150 of these patients (75 with an HCV code; 75 with a nonspecific viral hepatitis code) to collect information on HCV and other hepatitis diagnoses. We determined the positive predictive value of the database's HCV diagnostic codes and its ability to identify the date of a new HCV diagnosis. RESULTS: Usable surveys were returned for 146 (97%) patients. Among 74 patients with an HCV code and questionnaire data, HCV was confirmed in 64 (positive predictive value, 86%; 95%CI, 77-93%). In 40 (63%), the first recorded diagnosis in THIN was within 30 days of the date reported in the questionnaire (median difference, 11 days; interquartile range, 0-362 days). Among 72 patients with a nonspecific viral hepatitis code, 16 (22%) had HCV, but manual review of the database's electronic records correctly identified 12/16 (75%). CONCLUSIONS: In THIN, the HCV-specific diagnostic codes are highly predictive of HCV infection. After manual review, few patients with a nonspecific viral hepatitis code were misclassified as having HCV infection. PMID- 19551701 TI - What do Korean American immigrants know about Alzheimer's disease (AD)? The impact of acculturation and exposure to the disease on AD knowledge. AB - OBJECTIVE: The present study examined knowledge of Alzheimer's disease (AD) and factors related to the disease knowledge among Korean American immigrants. METHODS: Two-hundred and nine middle-aged and older Korean American immigrants completed a self-administered questionnaire. RESULTS: Korean American immigrants had strong stigma about AD and interpreted the disease as a form of insanity. They considered memory loss and AD as a part of the aging process. They seriously lacked knowledge about the treatment, diagnosis and cause of AD. Those who were less acculturated and less exposed to AD were likely to have poorer AD knowledge. CONCLUSIONS: There is a need to expand AD education efforts among Korean American immigrants because the views of stigmatization and normalization of AD combined with certain Asian cultural norms can be barriers to early recognition and intervention. Our study suggests knowledge areas of concentration in AD education and a segment of the population that AD education should target. Further research on symptom knowledge by the disease stages and causal beliefs is needed to better understand the existing views about AD in Korean American immigrants. PMID- 19551700 TI - Time trends in the use of anti-hypertensive medications: results from the Multi Ethnic Study of Atherosclerosis. AB - BACKGROUND: Previous reports have suggested that new evidence of the comparative effectiveness of different medication classes from randomized controlled trials (RCTs) does not always alter treatment decisions for first-line anti-hypertensive therapy. OBJECTIVES: To evaluate the association of RCT evidence in December 2002 from the Anti-hypertensive and Lipid-Lowering treatment to prevent Heart Attack Trial (ALLHAT) on use of anti-hypertensive medications in a multi-ethnic cohort. METHODS: The Multi-Ethnic Study of Atherosclerosis (MESA) study, a prospective cohort study of 6814 adults from four ethnic groups, had four separate assessments of drug use. Users of anti-hypertensive medications at baseline were excluded. We evaluated temporal changes in the medication class reported by new users of anti-hypertensive medications. RESULTS: After the exclusion of anti hypertensive drug users at baseline, 32% of new users of anti-hypertensive drugs seen at exam 2 were prescribed a diuretic. The publication of ALLHAT was associated with a subsequent increase in the proportion of new users taking diuretics at exam 3 compared with exam 2 (relative risk (RR): 1.31; 95% confidence interval (CI): 1.09-1.59). After the report from ALLHAT, the proportion of users of diuretics seen at exam 3 rose to 44% (starting in 2004) and 39% in exam 4 (starting in 2005). This increase in the proportion of diuretic use among new users of anti-hypertensive medications declined slightly but could still be detected at exam 4 as compared to exam 2 (RR: 1.28; 95%CI: 1.04-1.57). CONCLUSIONS: The randomized trial evidence from the ALLHAT study was temporally associated with a moderate increase in diuretic use. PMID- 19551702 TI - Electrocardiographic monitoring for QT prolongation in patients treated with ziprasidone--a claims database approach. AB - PURPOSE: Drug-induced cardiac arrhythmia is a major safety concern for drugs with a potential to prolong the QT interval. The purpose of the study is to examine whether timely electrocardiographic (ECG) monitoring has been performed for patients treated with ziprasidone. METHODS: Out-patient pharmacy claims data abstracted from Taiwan's National Health Insurance Research Database were used to identify patients treated with ziprasidone. Based on the dataset sorted by the encrypted patient identifier and by date, the rates of ECG performed before and during treatment were examined. The intervals between treatment beginning and ECG examination during therapy were calculated. RESULTS: Among 4789 patients ever treated with zipasidone, 229 (4.8%) had ECG performed before treatment. Among 2052 patients treated with a longer duration of ziprasidone, 394 (19.2%) had ECG performed during treatment. They included 64 (3.1%) patients who had it performed within the first 30 days of treatment, 124 (6.0%) within 60 days and 178 (8.7%) within 90 days. The mean interval was 157.4 days (95% confidence interval (CI), 143.0-171.8) between treatment beginning and the first on-therapy ECG examination and was 210.4 days (95%CI, 178.6-242.2) between treatment beginning and the second ECG. CONCLUSIONS: Low rates of ECG examination before and during therapy were noted among patients treated with ziprasidone. It took significant length of time to perform ECG monitoring after treatment was started. Although the patients studied had few risk factors for cardiac arrhythmia, ECG monitoring for patients using ziprasdione could be improved. PMID- 19551703 TI - Disagreement in preference for residential care between family caregivers and elders is greater among cognitively impaired elders group than cognitively intact elders group. AB - OBJECTIVES: This study examined the predictive factors of preference for residential care in cognitively intact and impaired elders and their family caregivers. It was hypothesized that disagreement in preference for residential care between the elders and their caregivers was greater in the cognitively impaired. METHODS: A cross-sectional survey was conducted during June 2007 to March 2008 in Hong Kong, and 707 community-dwelling elders aged 65 and above and 705 family caregivers were interviewed. Cognitively impaired elders were over sampled to give reliable estimates for that sub-group. A structural questionnaire was used to collect data on preference for residential care and potential factors. Logistic regression was used to identify the predictors. RESULTS: More cognitively impaired elder-caregiver dyads (37.4%) had disagreement in preference for residential care than cognitively intact elder-caregiver dyads (20.5%) (p < .001). From the elders' perspective, less preference for residential care was associated with cognitive impairment, whereas greater preference was associated with depression (for cognitively intact elders), more usage of community service and functional impairment. From the caregivers' perspective, greater preference for residential care was associated with greater caregiver burden, or care recipients having cognitive or functional impairment, or more usage of community services. CONCLUSIONS: Cognitively intact elders were more likely to indicate preference for residential care than cognitively impaired elders. Elders, both cognitively intact and impaired, were less likely than their caregivers to indicate preference for residential care. Disagreement in preference for residential care between the elders and their caregivers was larger for the cognitively impaired group than the cognitively intact group. PMID- 19551705 TI - Modified electrophoretic and digestion conditions allow a simplified mass spectrometric evaluation of disulfide bonds. AB - Proper formation of disulfide bonds in proteins is a prerequisite to their stability and function. Information on disulfide pattern may therefore serve as an indication of the proper folding of recombinant proteins, and can also be used in protein homology modeling for the purpose of structure refinement. Protein handling and digestion at basic pH leads to disulfide bond scrambling. That is why the samples are usually treated and digested at low pH where no scrambling occurs. Unfortunately, the specific proteases used in protein research are active at high pH values. Here, we present a complete sample handling protocol, which allows processing of disulfide containing proteins at basic pH. We modified the standard SDS gel electrophoresis and protein digestion conditions by the addition of an oxidative agent, cystamine. This modification prevented disulfide scrambling, which we otherwise observed in the samples handled according to the general protocol. Lysozyme from hen egg was used as a model protein for the development of the method. We then applied our protocol to human leukocyte antigen CD69, for which the disulfide bonding is known, but only for its monomeric form. In addition, the disulfide arrangement was then 'de novo' identified in the recombinant murine leukocyte receptor NKR-P1A and in the larger glycosylated proteins beta-N-acetylhexosaminidases from Aspergillus oryzae and Penicillium oxalicum. PMID- 19551706 TI - Screening for depression in elderly medical inpatients from rural area of Norway: prevalence and associated factors. AB - AIM: The present investigation screened for depression in order to assess the prevalence of depression and to study the associated factors with depression in elderly medically hospitalised patients from a rural area in Norway. METHODS: A cross-sectional study evaluated 484 (243 women) elderly medical inpatients with age range 65-101 (mean 80.7) years between September 2006 and August 2008 and used the Hospital Anxiety and Depression scale (HAD), Montgomery and Asberg Depression Rating Scale, the Mini-Mental State Examination, Lawton and Brody's scale for self-maintaining and instrumental activities of daily living. RESULTS: The prevalence of current depression, depression score > or =8 at HAD, was for the total sample 10% of whom 78% was previously not diagnosed as having depression. The odds for depression were decreased for women aged 80 years or more while for men at the same age strata it was increased threefold. Age adjusted logistic regression analyses demonstrated an increased odds for depression for those who were in need of nursing assistance before hospitalisation, had lower level of physical functioning, had clinical anxiety symptoms and had higher number of medicaments at inclusion time. CONCLUSION: The prevalence of depression in medical hospitalised elderly from rural areas was lower than in most other hospital studies. However, most patients with depression were not previously recognised as being depressed. PMID- 19551708 TI - Development of cardiac form and function in ectothermic sauropsids. AB - Evolutionary morphologists and physiologists have long recognized the phylogenetic significance of the ectothermic sauropsids. Sauropids have been classically considered to bridge between early tetrapods, ectotherms, and the evolution of endotherms. This transition has been associated with many modifications in cardiovascular form and function, which have changed dramatically during the course of vertebrate evolution. Most cardiovascular studies have focused upon adults, leaving the development of this critical system largely unexplored. In this essay, we attempt a synthesis of sauropsid cardiovascular development based on the limited literature and indicate fertile regions for future studies. Early morphological cardiovascular development, i.e., the basic formation of the tube heart and the major pulmonary and systemic vessels, is similar across tetrapods. Subsequent cardiac chamber development, however, varies considerably between developing chelonians, squamates, crocodilians, and birds, reflected in the diversity of adult ventricular structure across these taxa. The details of how these differences in morphology develop, including the molecular regulation of cardiac and vascular growth and differentiation, are still poorly understood. In terms of the functional maturation of the cardiovascular system, reflected in physiological mechanisms for regulating heart rate and cardiac output, recent work has illustrated that changes during ontogeny in parameters such as heart rate and arterial blood pressure are somewhat species-dependent. However, there are commonalities, such as a beta-adrenergic receptor tone on the embryonic heart appearing prior to 60% of development. Differential gross morphological responses to environmental stressors (oxygen, hydration, temperature) have been investigated interspecifically, revealing that cardiac development is relatively plastic, especially, with respect to change in heart growth. Collectively, the data assembled here reflects the current limited morphological and physiological understanding of cardiovascular development in sauropsids and identifies key areas for future studies of this diverse vertebrate lineage. PMID- 19551707 TI - Associations between stroke risk and cognition in normal aging and Alzheimer's disease with and without depression. AB - BACKGROUND: Stroke risk factors have been increasingly implicated in the development of age-related cognitive decline, the spectrum of vascular cognitive impairment, and, more recently, Alzheimer's disease (AD). In addition, depression and the apolipoprotein (APOE) epsilon4 allele have been reported to influence the association between stroke risk and cognition. However, few studies have described the relations among stroke risk, cognition, and APOE genotype in AD, and the findings have been equivocal. METHODS: Thirty cognitively normal older adults, 30 AD patients with depression, and 30 AD patients without depression were administered a comprehensive neuropsychological battery measuring several domains including memory, attention, language, visuospatial skills, executive functions, and speed of information processing. The Framingham Stroke Risk Profile (FSRP), a validated scale that was developed to predict 10-year probability of stroke, was used to quantify stroke risk burden. RESULTS: AD patients with depression demonstrated greater stroke risk burden relative to the cognitively normal group and, across all participants, increased stroke risk was associated with poorer performance on memory and processing speed measures. Moreover, stroke risk accurately predicted AD diagnosis. Notably, there were no significant differences in stroke risk or cognitive performance between the AD participants with depression and those without depression. CONCLUSION: Given that many markers of stroke risk are modifiable or treatable, our findings have implications for assessment, prevention, and treatment of cognitive decline. PMID- 19551709 TI - Living in fast-flowing water: morphology of the gastromyzophorous tadpole of the bufonid Rhinella quechua (R. veraguensis group). AB - We describe the bufonid gastromyzophorous tadpoles of Rhinella quechua from montane forest streams in Bolivia. Specimens were cleared and stained, and the external morphology, buccopharyngeal structures, and the musculoskeletal system were studied. These tadpoles show a combination of some traits common in Rhinella larvae (e.g., emarginate oral disc with large ventral gap in the marginal papillae, labial tooth row formula 2/3, prenarial ridge, two infralabial papillae, quadratoorbital commissure present, larval otic process absent, mm. mandibulolabialis superior, interhyoideus posterior, and diaphragmatopraecordialis absent, m. subarcualis rectus I composed of three slips), some traits apparently exclusive for the described species of the R. veraguensis group (e.g., second anterior labial tooth row complete, lingual papillae absent, adrostral cartilages present), and some traits that are shared with other gastromyzophorous tadpoles (e.g., enlarged oral disc, short and wide articular process of the palatoquadrate, several muscles inserting on the abdominal sucker). In the context of the substantial taxonomic and nomenclatural changes that the former genus Bufo has undergone, and despite the conspicuous morphological differences related to the presence of an abdominal sucker, the larval morphology of R. quechua supports including it in the genus Rhinella and placing it close to species of the R. veraguensis assemblage. PMID- 19551710 TI - Effect of vitrification on mitochondrial distribution and membrane potential in mouse two pronuclear (2-PN) embryos. AB - The present study was designed to investigate the effect of vitrification on mitochondrial distribution, membrane potential (Deltapsi) and microtubule distribution in mouse 2-PN embryos, as well as to document the relationship between mitochondrial distribution and developmental ability of those embryos. Mitochondrial distribution was examined by fluorescence microscopy technology. Results indicated that: (1) The rate of mitochondrial ring formation around pronuclei in vitrified 2-PN embryos was significantly lower than in fresh ones (67.3 +/- 3.0% vs. 84.9 +/- 3.1%) (P < 0.05). (2) Blastocyst development rate of vitrified 2-PN embryos without mitochondrial rings (61.7 +/- 4.5%) was significantly lower than that of vitrified embryos with mitochondrial rings (82.1 +/- 2.8%). (3) Following staining by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl imidacarbo-cyanine iodide (JC-1), most red-colored mitochondria (high Deltapsi) were distributed peripherally around pronuclei and along cell membranes of fresh 2-PN embryos. Conversely, red-colored mitochondria were greatly diminished in vitrified embryos, with green mitochondria (low Deltapsi) evenly distributed throughout the cytoplasm. The proportion of fresh 2-PN embryos with obvious aggregation of high Deltapsi mitochondria (84.2 +/- 2.2%) was significantly higher than that of vitrified embryos (26.7 +/- 3.0%) (P < 0.05). (4) The proportion of fresh embryos with microtubules distributed around pronuclei (83.5 +/- 3.4%) was similar to that of vitrified embryos (74.7 +/- 2.5%). In conclusion, vitrification affected mitochondrial distribution and decreased the mitochondrial membrane potential in mouse 2-PN embryos, events which may affect subsequent developmental viability of such embryos. PMID- 19551711 TI - Behavioral treatment of substance abuse in schizophrenia. AB - Co-occurring substance use disorders are highly prevalent among individuals with schizophrenia and other serious and persistent mental illnesses (SPMI) and are associated with clinically significant consequences. A multifaceted behavioral treatment called Behavioral Treatment for Substance Abuse in Serious and Persistent Mental Illness (BTSAS) can reduce substance abuse in persons with SPMI. The key treatment strategies in BTSAS include a urinalysis contingency, short-term goal setting, training in drug refusal skills, psychoeducation about the impact of drug use, and relapse prevention training. A case example illustrating the application of BTSAS is presented and relevant clinical issues are discussed. PMID- 19551712 TI - Familial recurrence of urethral stenosis/atresia. AB - BACKGROUND: We report the familial recurrence of urethral stenosis/atresia in two sibling fetuses with bladder outlet obstruction, severe oligohydramnios, and pulmonary hypoplasia. Urethral obstruction in the fetus, when severe, results in a dilated urinary bladder (megacystis) and associated urinary anomalies (hydroureter, hydronephrosis, renal dysplasia). Distention of the fetal abdomen, the result of megacystis or urinary ascites, leads to stretching and eventually hypoplasia or even absence of abdominal muscles. CASES: This constellation of findings, known by a variety of terms including "prune belly" syndrome, is associated with a variety of urethral changes, including posterior urethral valves and urethral stenosis/atresia. One fetus manifested unilateral postaxial polydactyly of the left hand. CONCLUSIONS: A microdeletion of 6p25.3, identified in mother and one fetus, is not associated with a gene known to be involved in urethral development and therefore of unknown significance. PMID- 19551713 TI - Regarding the need, or lack thereof, of a comparator group for pregnancy registries. PMID- 19551714 TI - New and improved: the role of text augmentation and the application of response interpretation standards (coding schemes) in a final iteration of birth defects warnings development. AB - BACKGROUND: Several birth defects warning symbols identified as most successful in an earlier study (Mayhorn and Goldsworthy, 2007) were further modified and then evaluated within a nationally distributed field trial (n = 2773). The purpose for the current research was to determine whether symbol warning components could be improved further, whether the addition of text enhanced comprehension uniformly across symbols, and whether results varied by the application of different interpretation standards (coding schemes). METHOD: A total of 11 warning labels were examined: four new symbols plus the existing baseline symbol, each in versions with and without text, plus a text-only condition. Participant interpretation accuracy and preferences were assessed during face-to-face interview sessions. RESULTS: For symbol-only conditions, several candidate symbols outperformed the existing symbol, one substantially so. The effect of adding text to symbols varied significantly by symbol. Symbol plus text and text-only conditions performed equivalently, generally exceeded symbol only conditions, and often surpassed the American National Standards Institute benchmark of 85% accurate interpretation. CONCLUSIONS: The research effort has identified a teratogen symbol and warning that outperforms the one currently in use. The effort has also identified important pragmatic and conceptual issues that should inform future work to improve medication labeling and other hazard communication. PMID- 19551715 TI - Folding in solution of the C-catalytic protein fragment of angiotensin-converting enzyme. AB - Angiotensin-converting enzyme (ACE) is a key molecule of the renin-angiotensin aldosterone system which is responsible for the control of blood pressure. For over 30 years it has become the target for fighting off hypertension. Many inhibitors of the enzyme have been synthesized and used widely in medicine despite the lack of ACE structure. The last 5 years the crystal structure of ACE separate domains has been revealed, but in order to understand how the enzyme works it is necessary to study its structure in solution. We present here the cloning, overexpression in Escherichia coli, purification and structural study of the Ala(959) to Ser(1066) region (ACE_C) that corresponds to the C-catalytic domain of human somatic angiotensin-I-converting enzyme. ACE_C was purified under denatured conditions and the yield was 6 mg/l of culture. Circular dichroism (CD) spectroscopy indicated that 1,1,1-trifluoroethanol (TFE) is necessary for the correct folding of the protein fragment. The described procedure can be used for the production of an isotopically labelled ACE(959-1066) protein fragment in order to study its structure in solution by NMR spectroscopy. PMID- 19551717 TI - Ontogeny of the androgen receptor expression in the fetal and postnatal testis: its relevance on Sertoli cell maturation and the onset of adult spermatogenesis. AB - From fetal life to adulthood, the testis evolves through maturational phases showing specific morphologic and functional features in its different compartments. The seminiferous cords contain Sertoli and germ cells, surrounded by peritubular cells, and the interstitial tissue contains Leydig cells and connective tissue. Sertoli cells secrete anti-Mullerian hormone (AMH), whereas Leydig cells secrete androgens. In the fetal and early postnatal testis, Leydig cells actively secrete androgens. Sertoli cells are morphologically and functionally immature--e.g., they secrete high levels of AMH--and germ cells proliferate by mitosis but do not enter meiosis. During infancy and childhood, Leydig cells regress and testosterone secretion declines dramatically. Sertoli cells remain immature and spermatogenesis is arrested at the premeiotic stage. At puberty, Leydig cells differentiate again, and testosterone concentration increases and provokes Sertoli cell maturation--e.g., down-regulation of AMH expression--and germ cells undergo meiosis, the hallmark of adult spermatogenesis driving to sperm production. An intriguing feature of testicular development is that, although testosterone production is as active in the fetal and early postnatal periods as in puberty, Sertoli cells and spermatogenesis remain immature until pubertal onset. Here, we review the ontogeny of the androgen receptor expression in the testis and its impact on Sertoli cell maturation and the onset of pubertal spermatogenesis. We show that the absence of androgen receptor expression in Sertoli cells underlies a physiological stage of androgen insensitivity within the male gonad in the fetal and early postnatal periods. PMID- 19551718 TI - Prior use of extreme weight-control behaviors in a community sample of women with binge eating disorder or subthreshold binge eating disorder: a descriptive study. AB - OBJECTIVE AND METHOD: The prior occurrence of regular extreme weight-control behaviors was examined in a community sample of women (n = 27) with binge eating disorder (BED) or subthreshold BED. RESULTS: Approximately two thirds of participants (65.4%) reported the prior use of either purging at least weekly or nonpurging behaviors three or more times per week, for a period of 3 months or more, whereas 38.5% of participants reported either purging at least twice weekly or nonpurging behaviors five or more times per week. Comparatively few participants (11.1%) had ever met formal diagnostic criteria for BN. DISCUSSION: Considerable overlap between disorders characterized by binge eating in the absence of extreme weight-control behaviors and those characterized by extreme weight-control behaviors in the absence of binge eating may be apparent when a longitudinal perspective is taken. Caution needs to be exercised in drawing conclusions concerning the extent of this overlap based on any one operational definition of the term "regular extreme weight-control behaviors". PMID- 19551723 TI - Chemical constituents of plants from the genus Blumea. PMID- 19551724 TI - Chemical constituents of plants from the genus Incarvillea. PMID- 19551725 TI - Cationic porphyrins and analogues as new DNA topoisomerase I and II inhibitors. AB - A series of cationic porphyrins, and analogues such as cationic corroles and phthalocyanines were found to have biological activities towards topoisomerases I and II in vitro. Cationic porphyrins and phthalocyanines do not induce Topo I-DNA covalent complexes but inhibit topoisomerase I by direct binding to DNA, which limits topoisomerase I access to the DNA substrate. The lowest concentration where an inhibition effect is clearly visible of some derivatives is between 0.1 and 0.6 microM. Furthermore, some complexes were found to inhibit the activity of the topoisomerase II. PMID- 19551726 TI - Antifungal activity of alkaloids from the seeds of Chimonanthus praecox. AB - Two alkaloids, D-calycanthine (1) and L-folicanthine (2), were isolated from the active MeOH extract of the seeds of Chimonanthus praecox LINK. The structures of the two compounds were established by (1)H- and (13)C-NMR, and MS (FAB, ESI) analyses. In the in vitro tests, compounds 1 and 2 showed significant inhibitory activities against five plant pathogenic fungi Exserohilum turcicum, Bipolaris maydis, Alternaria solani, Sclerotinia sderotiorum, and Fusarium oxysportium, among which B. maydis was found to be the most susceptible to 1 with an EC(50) value of 29.3 microg/ml, followed by S. sderotiorum to 2 with an EC(50) value of 61.2 microg/ml. To our knowledge, this is the first report of isolation and LC/MS/MS identification as well as of antifungal properties of these alkaloids from the seeds of this plant. PMID- 19551727 TI - Synthesis and biological evaluation of asiatic acid derivatives as inhibitors of glycogen phosphorylases. AB - Twenty-four asiatic acid derivatives have been synthesized and biologically evaluated as inhibitors of glycogen phosphorylase (GP). Within this series of compounds, asiatic acid benzyl ester (23; IC(50)=3.8 microM) exhibited more potent activity than its parent compound 1 (IC(50)=17 microM). SAR Analysis showed that asiatic acid (1) possessing a 2alpha-OH function exhibited more potent GP inhibitory activity than eriantic acid B (27) which possesses a 2beta OH function. Further lead optimization based on 1 is needed to find more effective asiatic acid derivatives as antidiabetic agents with protective effects against ischemic diabetic complications. PMID- 19551729 TI - Flower and root oils of the tunisian Daucus carota L. ssp. maritimus (Apiaceae): integrated analyses by GC, GC/MS, and 13C-NMR spectroscopy, and in vitro antibacterial activity. AB - The essential oils from flowers and roots of Daucus carota L. ssp. maritimus were obtained by hydrodistillation and analyzed by combination of GC, GC/MS, and (13)C NMR. The chemical composition of the flower and root oils of this subspecies is reported here for the first time. Thirty-two and six compounds were identified in flower and root oils, respectively. A remarkable difference was found between the constituent percentages of the two organs. The chemical composition of the essential oil from flowers was characterized by a high proportion of monoterpene hydrocarbons (68.4%) and belonged to the sabinene (4; 51.6%) chemotype. The oxygenated monoterpenes represented the second major fraction of the same oil among which terpinen-4-ol (11.0%) was the predominant compound. Root oil exhibited a surprisingly different composition. Phenolic derivatives (76.3%), myristicin (31; 29.7%), and dillapiole (32; 46.6%) constituted the main fraction. The antibacterial effect resulted in the inhibition of a series of common human pathogenic bacteria, and of some clinically and environmentally isolated strains with significant MIC and MBC values. PMID- 19551728 TI - Phenolic constituents from Balanophora laxiflora with DPPH radical-scavenging activity. AB - Balanophora laxiflora HEMSL. (Balanophoraceae), a dioeciously parasitic plant, has been used as a tonic and for sobering up from drunk by the local people of Yunnan province, China. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay on the acetone extract of the fresh female plant of B. laxiflora displayed considerable radical-scavenging activity (SC(50)=16.4 microg/ml). Further purification of the extract led to the isolation of two new phenolic glycosides, balaxiflorins A and B (1 and 2, resp.), together with 17 known phenolic compounds including 3 phenylpropanoids, 3, 17-18, four lignans, 4-7, nine hydrolyzable tannins, 8-16, and gallic acid (19). Their structures were determined by detailed spectroscopic analysis. The free-radical-scavenging activity of the isolated compounds was examined by DPPH assay. PMID- 19551730 TI - Immunological adjuvant effect of a water-soluble polysaccharide, CPP, from the roots of Codonopsis pilosula on the immune responses to ovalbumin in mice. AB - In this study, one water-soluble polysaccharide, CPP, was purified from the root of Codonopsis pilosula. The immunomodulatory effect and the adjuvant potential of CPP on the cellular and humoral immune response of ICR mice against ovalbumin (OVA) were investigated. CPP was shown not to be lethal in vivo for mice in doses ranging from 0.5 to 4 mg. ICR Mice were immunized subcutaneously with 0.1 mg of OVA alone or with 0.1 mg of OVA dissolved in saline-containing aluminum hydroxide gel (Alum) (0.2 mg), QuilA (0.01 and 0.02 mg) or CPP (0.5, 1 or 2 mg) on days 1 and 15. Two weeks later (day 28), concanavalin A (ConA)-, lipopolysaccharide (LPS)-, and OVA-stimulated splenocyte proliferation, and OVA-specific serum antibodies were measured. CPP significantly enhanced the ConA-, LPS-, or OVA induced splenocyte proliferation in the OVA-immunized mice especially at a dose of 1 mg (P<0.05 or P<0.01). The OVA-specific IgG, IgG1, and IgG2b antibody levels in serum were also significantly enhanced by CPP compared with OVA control group (P<0.05 or P<0.01). The results suggest that CPP could be a safe efficacious adjuvant for use in vaccines against both pathogens and cancer. PMID- 19551731 TI - Susceptibility of Aedes aegypti and Culex quinquefasciatus Larvae to gedunin related limonoids. AB - The major non-azadirachtin limonoids such as gedunin (1), epoxyazadiradione (3), nimbocinol (4), and nimolicinol (5) from Azadirachta indica A. Juss ('neem') and their derivatives were evaluated for their toxic action against fourth instar larvae of Aedes aegypti L. and Culex quinquefasciatus Say. Gedunin exhibited 100% toxic action against both the mosquito larvae at 50 and 10 ppm. Epoxyazadiradione and epoxynimolicinol also showed significant toxicities (> or =50%) against larvae of both mosquito species at 50 ppm. These neem limonoids can have benefits in mosquito-control programs. PMID- 19551732 TI - Acylated oleanane-type triterpene saponins with acceleration of gastrointestinal transit and inhibitory effect on pancreatic lipase from flower buds of chinese tea plant (Camellia sinensis). AB - The MeOH extract and its BuOH-soluble fraction (crude saponin fraction) from the flower buds of Chinese tea plant (Camellia sinensis (L.) O. KUNTZE; Fujian Province) were found to exhibit accelerating effects on gastrointestinal transit in mice and inhibitory effects against pancreatic lipase. From the BuOH-soluble fraction, three new acylated oleanane-type triterpene oligoglycosides, chakasaponins I, II, and III (1-3, resp.), were isolated together with 13 known compounds. The chemical structures 1-3 were elucidated on the basis of chemical and physicochemical evidence. Compounds 1-3 showed accelerating effects on gastrointestinal transit in mice and inhibitory effects against porcine pancreatic lipase (IC(50)=150-530 microM). PMID- 19551733 TI - Stemucronatoside L, a pregnane glycoside from the roots of Stephanotis mucronata, inhibits Th1/Th2 immune responses in vitro. AB - Stemucronatoside L (SML), isolated from Stephanotis mucronata, could suppress the activation of T cells in vitro. However, the mechanisms responsible for its immunosuppressive activity remain poorly understood. The purpose of this study was to investigate whether SML could suppress Th1/Th2 immune responses and to characterize the cellular mechanisms involved. Effects of SML on T-lymphocyte subsets and the production of Th1 cytokines IL-2 and IFN-gamma, and Th2 cytokines IL-4 and IL-10 from ConA-stimulated mice splenocytes were detected by flow cytometric analysis and ELISA method, respectively. Furthermore, effects of SML on mRNA expression level of Th1/Th2 cytokines and transcription factors T-bet and GATA-3 were evaluated by RT-PCR analysis. SML not only significantly decreased the percentage of CD4(+) T cells and the CD4(+)/CD8(+) ratio, but reduced the production of Th1/Th2 cytokines in a concentration-dependent manner. The mRNA expression levels of Th1/Th2 cytokines and transcription factors (T-bet and GATA 3) were also suppressed by SML. These results suggested that SML could simultaneously inhibit Th1/Th2 immune responses by suppressing gene expression of Th1/Th2 cytokines and transcription factors. PMID- 19551734 TI - Secondary metabolites and cytotoxic activities from the stem bark of Zanthoxylum nitidum. AB - A dihydrobenzo[c]phenanthridine alkaloid, epizanthocadinanine A (1), together with 27 known compounds, including eight benzo[c]phenanthridines, i.e., oxynitidine (2), oxyavicine (3), oxychelerythrine (4), dihydrochelerythrine (5), 6-acetonyldihydrochelerythrine (6), norchelerythrine (7), decarine (8), and arnottianamide (9); two 2-quinolones, i.e., flindersine (10) and 4-methoxy-1 methyl-2-quinolone (11); two furoquinolines, i.e., skimmianine (12) and gamma fagarine (13); three aporphines, i.e., liriodenine (14), N-acetyldehydroanonaine (15), and N-acetylanonaine (16); six lignans, i.e., sesamin (17), episesamin (18), piperitol-3,3-dimethylallyl ether (19), xanthoxylol-3,3-dimethylallyl ether (20), savinin (21), and 2,3-bis(3,4-methylenedioxybenzyl)but-2-en-4-olide (22); three terpenoids, i.e., alpha-cadinol (23), anticopalol (24), and spathulenol (25); one coumarin, i.e., aesculetin dimethyl ether (26); and two steroids, i.e., beta-sitosterol (27) and beta-sitostenone (28) were isolated from the stem bark of Zanthoxylum nitidum. Their structures were elucidated on the basis of extensive 1D- and 2D-NMR as well as MS analyses. Moreover, the recently reported structures 2'-4' of rhoifolines B and A, and '8-methoxynorchelerythrine', resp., isolated as new compounds from Z. rhoifolium and Z. nitidum, resp., could be assigned the revised structures 2-4 by reinvestigation of the spectroscopic data. In addition, the cytotoxicity of the isolates was evaluated on the MCF-7, NCI H460, and SF-268 cell lines. Among these isolates, liriodenine (14) was the most active compound against the MCF-7, NCI-H460, and SF-268 cell lines with IC(50) values of 2.19, 2.38, and 3.19 microg/ml, resp. PMID- 19551735 TI - Two new unprecedented acetonyl-bearing sesquiterpenes from the hainan sponge Dysidea fragilis. AB - Two new acetonyl-bearing sesquiterpenes, dysifragilisins A and B (1 and 2, resp.), have been isolated from the Hainan marine sponge Dysidea fragilis. The structures of the new compounds were elucidated on the basis of detailed spectroscopic analyses and by comparison of their NMR data with those of structurally related compounds. A plausible biogenetic pathway is proposed for the origin of compounds 1 and 2. PMID- 19551736 TI - Effective insect repellent formulation in both surfactantless and classical microemulsions with a long-lasting protection for human beings. AB - The aim of this work is to develop a new generation of repellent products with a long-lasting protection based on a natural component, para-menthane-3,8-diol (PMD). The active is first rendered soluble in a surfactantless microemulsion (H(2)O/(i)PrOH/PMD) and then in classical microemulsions. The presence of self associated nanostructures is detected by dynamic light scattering (DLS). A synergetic system of surfactants (Cremophor) RH40 and Texapon N70) is used. Additionally, 2-ethylhexane-1,3-diol and ethyl (-)-(S)-lactate are incorporated. The final product contains, as main components, 46% of H(2)O, 25% of (i)PrOH, 20% of non-H(2)O-soluble PMD, and only 4% of surfactants. Investigations of lasting protection on human volunteers are carried out using a cage test bioassay protocol and Aedes aegypti mosquitoes. A complete protection of 315 min is found on the test persons using the surfactantless microemulsion. An extension is observed with the final formulation to reach a mean of complete protection of 385 min. This study demonstrates that alternative formulations using a natural active instead of synthetic chemicals like N,N-diethyl-m-methylbenzamide (DEET) can be efficient for human protection against mosquitoes. PMID- 19551737 TI - Anti-inflammatory, antiproliferative, and radical-scavenging activities of tolfenamic acid and its metal complexes. AB - Some new complexes of tolfenamic acid (=2-[(2-methyl-3-chlorophenyl)amino]benzoic acid; Htolf) with potentially interesting biological activities are described. The complexes [Mn(tolf)(2)(H(2)O)(2)], [Co(tolf)(2)(H(2)O)(2)], [Ni(tolf(2)(H(2)O)(2)], [Cu(tolf)(2)(H(2)O)](2), and [Zn(tolf)(2)(H(2)O)] were prepared by the reaction of tolfenamic acid, a potent anti-inflammatory drug, with metal salts. The radical-scavenging activities of the complexes were evaluated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging assay. Their ability to inhibit soybean lipoxygenase, beta-glucuronidase, and trypsin-induced proteolysis was studied. Their inhibitory effects on rat paw edema induced by carrageenin was studied and compared with those of tolfenamic acid. The complex [Zn(tolf)(2)(H(2)O)] exhibited the strongest in vivo inhibitory effect at 0.1 mm/kg Body Weight (BW; 93.0+/-0.9%), superior than the inhibition induced by tolfenamic acid at the same molar dose (76.0+/-0.9%). Tolfenamic acid and its metal complexes have been evaluated for antiproliferative activity in vitro against the cells of three human cancer cell lines, MCF-7 (breast cancer cell line), T24 (bladder cancer cell line), and A-549 (non-small cell lung carcinoma), and a mouse fibroblast L-929 cell line. The complexes [Mn(tolf)(2)(H(2)O)(2)] and [Cu(tolf)(2)(H(2)O)](2) have shown selectivity against T24 cell line. The IC(50) values of these two complexes against T24 cancer cell lines are in a micromolar range similar or better to that of the antitumor drug cisplatin. PMID- 19551739 TI - Liquid chromatographic determination of benomyl in water samples after dispersive liquid-liquid microextraction. AB - A dispersive liquid-liquid microextraction (DLLME) method combined with solvolysis reaction for extraction of the carbamate fungicide benomyl as carbendazim from water samples is described. The method is based on the extraction of benomyl from acidified sample solution and its conversion into carbendazim via solvolysis reaction with DMF as organic solvent. The proposed DLLME method was followed by HPLC with fluorimetric detection for determination of benomyl. The proposed method has good linearity (0.998) with wide linear dynamic range (0.01-25 mg/L) and low detection limit (0.0033 mg/L), making it suitable for benomyl determination in water samples. PMID- 19551740 TI - Doubly tethered tertiary amide linked and ionically bonded diproline chiral stationary phases. AB - Oligoproline chiral stationary phase (CSP) is a new class peptide chiral stationary phase. Many proline chiral stationary phases with different proline chain lengths and linkers have been prepared and evaluated. However, the doubly tethered and ionic type linkers have not been adequately investigated. In this study, covalently and ionically bonded chiral stationary phases with doubly tethered linker were prepared and characterized. The new covalently bonded doubly tethered diproline CSP was applied successfully to resolve various enantiomers of acidic, basic, and neutral compounds with phenyl, naphthyl, anthryl, or similarly sized groups. The enantiorecognition performances of singly and doubly tethered diproline CSPs were comparable. Variation of the type and content of organic modifiers in hexane or heptane mobile phase showed that the branch alcohols such as 2-propanol and 2-butanol, 1,2-dichloroethane, methyl tert-butyl ether, and ethyl acetate in the mobile phase enhanced chiral separation. End-capping on doubly tethered diproline CSP did not always improve the separation factor and resolution. Due to the rigid structure of the double tether, the enantioseparation ability of ionically bonded diproline CSP was well expressed to some analytes. PMID- 19551741 TI - Solid-phase extraction and residue determination of glyphosate in apple by ion pairing reverse-phase liquid chromatography with pre-column derivatization. AB - A new method for glyphosate residue determination in apple has been developed. A SPE cartridge was used to clean up the samples before derivatization. Glyphosate was derivatized with 4-chloro-3,5-dinitrobenzotrifluoride (CNBF) and quantified by reverse ion-pair liquid chromatography using cetyltrimethylammonium bromide (CTAB) as ion-pair reagent. In pH 9.5 H(3)BO(3)-Na(2)B(4)O(7) medium, the reaction of glyphosate with CNBF was complete after 30 min at 60 degrees C. The stability of the derivative on exposure to light at room temperature in methanol water was demonstrated. The labeled glyphosate was separated on a Kromasil C(18) column (250 x 4.6 mm, 5 microm) at room temperature and UV detection was applied at 360 nm. Separation was achieved within 15 min in gradient elution mode. The correlation coefficient for the method was 0.9998 at concentrations ranging from 0.1 to 50 microg/g. The calculated recoveries for glyphosate in apple were from 86.00 to 99.55%, and the relative standard deviations (n = 6) were from 1.43 to 6.32. The limit of detection was 0.01 microg/g for glyphosate in apple. PMID- 19551742 TI - Antiproliferation and apoptosis induced by evodiamine in human colorectal carcinoma cells (COLO-205). AB - Evodiamine (1), a biologically active alkaloid isolated from Evodia rutaecarpa (known in Chinese as Wu-Chu-Yu), has antioxidant, anti-inflammatory, and anticancer activities. It has recently been demonstrated that the cytotoxic activities of 1 might be due to its ability to inhibit cell growth and induce apoptosis. In this study, we investigated the effects of 1 on growth and apoptosis in COLO-205 cells by MTT assay, fluorescence microscopy, transmission electron microscopy, DNA fragmentation assay, flow cytometry, immunohistochemical analysis, Western blotting, and caspase-3 activity assay. Our data revealed that 1 could significantly inhibit COLO-205 cell proliferation in a dose-dependent manner, and 1-treated COLO-205 cells displayed typical morphological apoptotic characteristics and formation of DNA ladders in agarose gel electrophoresis. The COLO-205 cell cycle was arrested in G(2)/M phase by 1. Meanwhile, 1 increased the expression of Bax and p53, decreased the expression of Bcl-2, lowered the mitochondrial transmembrane potential, and induced the activation of caspase-3. These activities may contribute to the anticarcinogenic action of 1. PMID- 19551743 TI - In situ ethylation of organolead, organotin and organomercury species by bromomagnesium tetraethylborate prior to GC-ICP-MS analysis. AB - An analytical method for simultaneous in situ ethylation, of organolead, organotin and organomercury compounds in aqueous samples was developed using a new derivatisation agent, bromomagnesium tetraethylborate (BrMgEt(4)B). The determination of lead, tin and mercury compounds was done by species-specific isotope dilution, derivatisation and GC-inductively coupled plasma MS (GC-ICP-MS) or by GC-MS. The recovery and accuracy of the derivatisation were evaluated. The effect of pH and the relative quantity of derivatisation agent were studied. PMID- 19551744 TI - Capillary zone electrophoresis with field enhanced sample stacking as a tool for targeted metabolome analysis of adenine nucleotides and coenzymes in Paracoccus denitrificans. AB - The main aim of this work was to demonstrate the applicability of capillary zone electrophoresis in combination with field enhanced sample stacking in targeted metabolome analyses of adenine nucleotides--AMP, ADP, ATP, coenzymes NAD(+), NADP(+) and their reduced forms in Paracoccus denitrificans. Sodium carbonate/hydrogencarbonate buffer (100 mM, pH 9.6) with the addition of beta-CD at a concentration of 10 mM was found to be an effective BGE for their separation within 20 min. Besides this, special attention was paid to the development of the procedure for the extraction of specific metabolites from the bacterium P. denitrificans. This procedure was not only optimised to achieve the highest metabolite yields but also to obtain a sample that was fully compatible with the online preconcetration strategy used. The developed methodology was finally applied in a study of the bacterium P. denitrificans at various stages of the active respiratory chain. PMID- 19551745 TI - Rapid and efficient screening of adsorbent for oligopeptide using molecular docking and isothermal titration calorimetry. AB - A rapid and efficient method based on molecular docking and isothermal titration calorimetry (ITC) was developed to identify effective adsorbents for the target peptide Ser-Glu-Ala-Asp-His (SEADH). Preliminary screening of five candidate adsorbents using molecular docking revealed that three suitable structures (A1, A2, and A3) either with or without coordination of Zn(2+) should be effective. The three selected structures were then prepared and further screened by evaluating their affinities for the peptide SEADH using ITC. The screening results revealed that the adsorbent A2 coordinated with Zn(2+) was the best adsorbent, and subsequent static adsorption experiments confirmed the reliability of the screening method. Further ITC analysis, combined with molecular docking, was performed to provide the possible adsorption mechanism. PMID- 19551746 TI - An analysis of life-course smoking behavior in China. AB - With a total population of more than 1.3 billion people where more than 31% of adults smoke, China has become the world's largest producer and consumer of cigarettes. We adopt a life-course perspective to study the economics of smoking behavior in China. We use data from the China Health and Nutrition Survey (CHNS) to follow individuals over their whole lives and to analyze their decisions to both start and stop smoking. We extend the small but growing body of economic research on smoking in China. Our life-course approach emphasizes that current smoking participation reflects a decision to start and a series of past decisions to not quit. We explore how the determinants of smoking initiation differ from the determinants of smoking cessation. We find results, consistent with some previous empirical evidence, that Chinese smoking is not strongly related to the price of cigarettes. Based on our results, we offer some speculative hypotheses that, we hope, might guide future research on the economics of smoking in China. It seems especially useful to compare the broad patterns we document with the experiences of other countries. PMID- 19551747 TI - Health insurance and catastrophic illness: a report on the New Cooperative Medical System in rural China. AB - The overall goal of the paper is to understand the progress of the design and implementation of China's New Cooperative Medical System (NCMS) program between 2004 (the second year of the program) and 2007. In the paper we seek to assess some of the strengths and weaknesses of the program using a panel of national representative, household survey data that were collected in 2005 and early 2008. According to our data, we confirm the recent reports by the Ministry of Health that there have been substantial improvements to the NCMS program in terms of coverage and participation. We also show that rural individuals also perceive an improvement in service by 2007. While the progress of the NCMS program is clear, there are still weaknesses. Most importantly, the program clearly does not meet one of its key goals of providing insurance against catastrophic illnesses. On average, individuals that required inpatient treatment in 2007 were reimbursed for 15% of their expenditures. Although this is higher than in 2004, on average, as the severity of the illness (in terms of expenditures on health care) rose, the real reimbursement rate (reimbursement amount/total expenditure on medical care) fell. The real reimbursement rate for illnesses that required expenditures between 4000 and 10,000 yuan (over 10,000 yuan) was only 11% (8%). Our analysis shows that one of the limiting factors is constrained funding. PMID- 19551748 TI - Are services delivered by community health centers more cost-effective? Evidence from urban China. AB - China has introduced a system of community health centers (CHCs) to provide primary care. To test whether services provided by such centers are more cost effective than treatment at local higher-level hospitals, the study compared health outcomes and expenditures for patients with hypertension and diabetes mellitus in three cities. We hypothesized that treating patients in stable condition at CHCs is less costly than providing treatment in higher-level hospitals with no differences in health outcomes. Results indicate that daily drug and other medical expenditures were consistently equal or lower for patients seeking treatment in CHCs than for those treated in hospitals. Patients also saved time by visiting CHCs. Health outcomes, measured as mean arterial pressure for hypertension and plasma glucose for diabetes, were similar for patients seeking treatment in CHCs and hospitals in most cases. Results suggest that CHCs are more cost-effective than hospitals in treating chronic diseases. Findings may also indicate that those patients seeking care at hospitals have more serious- and therefore more expensive and time-consuming--conditions. Further empirical research is needed to clarify these results. PMID- 19551749 TI - Does per-diem reimbursement necessarily increase length of stay? The case of a public psychiatric hospital. AB - Payment methods can affect providers' behaviour and in turn influence the outcome of medical services. The per-diem reimbursement method is predicted to increase length of stay (LOS) and reduce daily expenditure. Using a Difference in Differences design, this study empirically examines the impact of changing from fee-for-service to per-diem reimbursement in a large psychiatric hospital in Beijing. Results show that the LOS did not increase but daily expenditure in fact increased. We provide several potential explanations for these puzzling findings, including the internal contracts between the hospitals and their physician staff among public hospitals in China. PMID- 19551750 TI - The Urban Resident Basic Medical Insurance: a landmark reform towards universal coverage in China. AB - As the latest government effort to reform China's health care system, Urban Resident Basic Medical Insurance (URBMI) was piloted in seventy-nine cities during the summer of 2007, following State Council Policy Document 2007 No. 20's guidelines. This study presents the first economic analysis of URBMI, following a national household survey in nine representative Chinese cities. The survey aimed to answer three questions: Who is covered by the plan? Who gains from the plan? Who is most satisfied with the plan? We have found that there is a U-shaped relationship between URBMI participation rate and income. That is, the extremely rich or poor are the most likely to participate. Those with any inpatient treatment last year or with any chronic disease are also more likely to enroll in URBMI, indicating adverse selection into participation. We have also found that in reducing financial barriers to care, URBMI most significantly benefits the poor and those with previous inpatient care. Finally, those participants in the bottom 20% of family incomes are happier with URBMI than are their more affluent counterparts. PMID- 19551751 TI - Health-seeking behavior and hospital choice in China's New Cooperative Medical System. AB - Since the dissolution of the Rural Cooperative Medical System at the end of the commune period, illness has emerged as a leading cause of poverty in rural China. To address the poor state of health care, the Chinese government unveiled the New Cooperative Medical System in 2002. Because local governments have been given significant control over program design, fundamental characteristics of the program vary from one county to the next. These differences may influence the decision to seek health care as well as the choice of hospital conditional on that initial decision. In this paper, we use a nested logit model to analyze household survey data from 25 counties to analyze the determinants of such health seeking behavior. We find that age, the share of household expenditures allocated to food consumption (a measure of relative income), and the presence of other sick people in the household negatively affect the decision to seek health care while disability has a positive influence. Further, conditional on seeking treatment, the reimbursement scheme in place in each county and the average daily expenditure associated with hospitalization strongly influence hospital choice. PMID- 19551752 TI - The New Cooperative Medical Scheme in rural China: does more coverage mean more service and better health? AB - This paper explores the impact of the New Cooperative Medical Scheme (NCMS), a newly adopted public health insurance program in rural China. Using a longitudinal sample drawn from the China Health and Nutrition Survey (CHNS), we employed multiple estimation strategies (individual fixed-effect models, instrumental variable estimation, and difference-in-differences estimation with propensity score matching) to correct the potential selection bias. We find that participating in the NCMS significantly decreases the use of traditional Chinese folk doctors and increases the utilization of preventive care, particularly general physical examinations. However, we do not find that the NCMS decreases out-of-pocket expenditure nor do we find that it increases utilization of formal medical service or improves health status, as measured by self-reported health status and by sickness or injury in the past four weeks. Our study indicates that despite the wide expansion of coverage, the impact of the NCMS is still limited. PMID- 19551753 TI - China's health system and its reform: a review of recent studies. AB - This paper provides a survey of the recent empirical research on China's 'old' health system (i.e. prior to the spate of reforms beginning in 2003). It argues that this research has enhanced our understanding of the system prior to 2003, in some cases reinforcing conclusions (e.g. the demand-inducement associated with perverse incentives) while in other cases suggesting a slightly less clear storyline (e.g. the link between insurance and out-of-pocket spending). It also concludes that the research to date points to the importance of careful evaluation of the current reforms, and its potential to modify policies as the rollout proceeds. Finally, it argues that the research on the pre-2003 system suggests that while the recently announced further reforms are a step in the right direction, the hoped-for improvements in China's health system will far more likely occur if the reforms become less timid in certain key areas, namely provider payments and intergovernmental fiscal relations. PMID- 19551754 TI - Economic analysis of China's health care system: turning a new page. PMID- 19551755 TI - Health economics in China. Foreword. PMID- 19551756 TI - Congenital abnormalities of 88 children born to mothers who attempted suicide with phenobarbital during pregnany: the use of a disaster epidemiological model for the evaluation of drug teratogenicity. AB - OBJECTIVE: To study the effect of large doses of phenobarbital used for a suicide attempt during pregnancy for fetuses to estimate the teratogenic potential of phenobarbital. DESIGN AND SETTING: Comparative analysis of exposed children and their unexposed sibs born to pregnant patients who attempted suicide during pregnancy and admitted to the toxicological inpatient clinic, Budapest, 1960 1993. STUDY PARTICIPANTS: Of 1044 self-poisoned pregnant women, 88 used phenobarbital for suicide attempt and delivered newborn babies. MAIN OUTCOME MEASURES: Structural birth detects, i.e., congenital abnormalities (CAs). RESULTS: Doses ranged between 400 and 3000 mg of phenobarbital in 88 pregnant women who attempted suicide with this drug during pregnancy and later delivered live-born babies. Twelve (13.6%) of 88 exposed children and eight (10.3%) of 78 sibs were affected with CAs (odds ratios (OR) with 95%CI: 1.4, 0.6-3.5). Of 88 exposed children, 34 were born to mothers who attempted suicide with phenobarbital between the 3rd and 12th postconceptional weeks, the critical period of most CAs; 3 had CAs (diaphragmatic defect, multiple CA, undescended testis), but only diaphragmatic defect might be associated with phenobarbital used for suicide attempt. CONCLUSIONS: The use of phenobarbital once but extremely large doses in non-epileptic pregnant women does not seem to pose a significant risk for CAs. Our experiences show the feasibility and benefits of the disaster epidemiological self-poisoning model for estimating human teratogenic risks of drug exposure. PMID- 19551758 TI - NSAID use and risk of leukaemia: a population-based case-control study. AB - PURPOSE: The use of non-steroidal anti-inflammatory drugs (NSAIDs) is associated with a reduced risk of developing colorectal and lung cancer. Studies suggesting similar associations in leukaemia have been small and underpowered. We have conducted a large population-based case-control study to determine whether the use of NSAIDs is associated with a reduced risk of acute and chronic leukaemias, and whether their use has any impact on survival in these patients. METHODS: We identified all the incident cases of leukaemia in 'The Health Improvement Network' (THIN) general practice dataset, along with at least four matched controls per case. We used conditional logistic regression to determine odds ratios for NSAID prescription rates and the risk of developing several leukaemia sub-types. We then used Cox regression to determine the association between NSAID prescription rate and risk of death in leukaemia. Hazard ratios were adjusted for gender, age at diagnosis, smoking status and Townsend Score. RESULTS: The risk of leukaemia overall appears to increase marginally with increased use of NSAIDs prior to diagnosis. This is not seen when individual leukaemia sub-types are examined, however, except perhaps in CLL where patients who had received 2-5 prescriptions/year are 34% more likely to be diagnosed with CLL than those who had not had any NSAID prescriptions (O.R. 1.34, p = 0.03, 95% C.I. 1.02-1.74). There was no statistically significant association between exposure to NSAIDs prior to leukaemia diagnosis and survival. CONCLUSION: The use of NSAIDs does not reduce the risk of developing leukaemia, nor do they improve survival. PMID- 19551757 TI - Hexabromocyclododecane decreases the lytic function and ATP levels of human natural killer cells. AB - This study investigates the effect of hexabromocyclododecane (HBCD) on the lytic function of human natural killer (NK) cells and on ATP levels in NK cells. NK cells are capable of lysing tumor cells, virally infected cells, and antibody coated cells. HBCD is a brominated cyclic alkane used primarily as an additive flame retardant. If HBCD interferes with NK cell function, this could increase risk of tumor development and/or viral infection. NK cells were exposed to various concentrations of HBCD for 24 and 48 h and 6 days before determining lytic function and ATP levels. ATP levels and lytic function were also determined in NK cells that were exposed to HBCD for 1 h followed by 24 and 48 h, and 6 days in HBCD-free media. The results indicated that exposure of NK cells to 10 microm HBCD for 24 h causes a very significant decrease in both NK cell lytic function and ATP levels (93.5 and 90.5%, respectively). Exposure of NK cells to 10 microm HBCD for 1 h followed by 24 h in HBCD-free media showed a progressive and persistent loss of lytic function (89.3%) as well as a decrease in ATP levels (46.1%). The results indicate that HBCD exposures decreased lytic function as well as ATP levels. However, a decrease in lytic function was not necessarily accompanied by a similar decrease in ATP. Importantly, these results also indicate that a brief (1 h) exposure to HBCD causes a progressive loss of lytic function over a 6 day period. PMID- 19551759 TI - cblC: advances in defining the MMACHC mutation spectrum. PMID- 19551760 TI - Genetic heterogeneity and cystic fibrosis. PMID- 19551761 TI - Polymer-peptide block copolymers - an overview and assessment of synthesis methods. AB - Incorporating peptide blocks into block copolymers opens up new realms of bioactive or smart materials. Because there are such a variety of peptides, polymers, and hybrid architectures that can be imagined, there are many different routes available for the synthesis of these chimera molecules. This review summarizes the contemporary strategies in combining synthesis techniques to create well-defined peptide-polymer hybrids that retain the vital aspects of each disparate block. Living polymerization can be united with the molecular-level control afforded by peptide blocks to yield block copolymers that not only have precisely defined primary structures, but that also interact with other (bio)molecules in a well defined manner. PMID- 19551762 TI - A safety and tolerability laboratory study of the combination of aripiprazole and topiramate in volunteers who drink alcohol. AB - OBJECTIVE: There are no reports examining the safety of taking both topiramate and aripiprazole together with alcohol. The ultimate aim for this research is to determine whether this combination is safe and is superior to either drug taken alone in reducing alcohol use in alcohol dependent patients. METHOD: This was an open-label trial. Thirteen heavy drinking participants not seeking treatment for alcoholism were randomized. Participants were titrated up to 300 mg of topiramate and 30 mg of aripiprazole a day over 35 days. Participants reported adverse events (AEs) daily alcohol use and participated in an alcohol challenge session (ACS). RESULTS: The eight participants who completed the study achieved the maximum doses of drugs. The AEs of the drugs would suggest that the AEs profile is broader but not additive. Alcohol use from the 28 days before screening to the seven days before the ACS was reduced (p = 0.08). CONCLUSION: There was no evidence that AEs of aripiprazole and topiramate are additive and can, therefore, be administered safely together with a modest amount of alcohol. There was also a trend for a reduction of alcohol use by participants. This finding has implications for further investigation of this combination of drugs for alcohol dependence. PMID- 19551763 TI - Melperone, an aytpical antipsychotic drug with clozapine-like effect on plasma prolactin: contrast with typical neuroleptics. AB - OBJECTIVE: To evaluate the effect of melperone, a butyrophenone with atypical antipsychotic properties, on plasma prolactin (PRL) concentrations compared with clozapine and typical neuroleptics. METHODS: Analysis of pre- and post-treatment PRL levels collected prospectively per protocol in a non-randomized study of 26 melperone-, 76 clozapine-, and 66 neuroleptic-treated patients with schizophrenia or schizoaffective disorder. Cross-sectional analysis of a larger sample of patients with PRL data was also performed. RESULTS: For males, post-treatment PRL levels were significantly higher in the typical neuroleptic group compared with the melperone (p = 0.0001) and clozapine (p = 0.0001) groups, with no significant difference between clozapine and melperone. For females, post-treatment PRL levels were significantly higher in the melperone group as compared to the clozapine group (p = 0.004). There were too few typical neuroleptic-treated females to permit analysis of this sample. However, the cross-sectional analysis of PRL data confirmed the results for melperone- and clozapine-treated females, and showed higher PRL levels in typical neuroleptic-treated females as compared with those who received melperone and clozapine. CONCLUSION: Melperone did not significantly increase PRL levels in male patients. However, melperone and typical neuroleptics caused increase in PRL levels in females. Further study of melperone's effects on PRL concentration is warranted. PMID- 19551765 TI - The relation between the specific absorption rate and electromagnetic field intensity for heterogeneous exposure conditions at mobile communications frequencies. AB - The relation between the incident electromagnetic field strength and both the whole-body and the local specific absorption rate (SAR) was investigated for typical heterogeneous exposure scenarios for frequencies relevant for mobile communication. The results were compared to results from plane wave exposure. Heterogeneous exposure arises from multiple path propagation of the electromagnetic waves to the location of interest. It is shown that plane wave exposure does not represent worst-case exposure conditions. When the electric field strength arising at plane wave exposure is compared to the electric field strength averaged over the volume of the human body occurring during multipath exposure, 12% of all heterogeneous cases examined represent worse exposure conditions than plane wave exposure for whole-body exposure at 946 MHz, 15% at 1840 MHz, and 22% at 2140 MHz. The deviation between plane wave and heterogeneous whole-body SAR ranges from -54% to 54%. For partial-body SAR averaged over 10 g of tissue, a range from -93% to 209% was found when comparing multiple wave exposure to single incoming plane waves. The investigations performed using the Visible Human as phantom showed that the basic restrictions are met as long as the reference levels are not exceeded. However, this must not be necessarily the case when different phantoms are used to perform similar investigations because recent studies demonstrated that reference levels might not be conservative when phantoms of children are used. Therefore, the results of this work indicate the need to extend the investigations to numerical simulations with additional human phantoms representing parts of the human population having different anatomy and morphology compared to the phantom used within the frame of this project. This also applies to phantoms of children. PMID- 19551764 TI - Effects of ectopic expression of human telomerase reverse transcriptase on immortalization of feather keratinocyte stem cells. AB - Normal somatic cells possess a finite life span owing to replicative senescence. Telomerase functions as a potential regulator of senescence in various cells. Expression level of human telomerase reverse transcriptase (hTERT) is correlated with telomerase activity and cellular immortalization. In this study, we investigated the effects of ectopic expression of hTERT on proliferation potential of chicken feather keratinocyte stem cells (FKSCs). We established FKSCs transduced with hTERT catalytic subunit fused with EGFP marker gene (hTERT EGFP-FKSCs). hTERT-EGFP-FKSCs had the great potential of proliferation in vitro and expressed kerainocyte stem cell markers integrin beta1 and CD49c. Keratin 15 and keratin 19, as native FKSCs, were also detected in hTERT-EGFP-FKSCs. By the analysis of fluorescent RT-PCR, western blotting and TRAP assay, hTERT-EGFP-FKSCs were positive for telomerase activity, in comparison with native FKSCs showing no telomerase activity. We demonstrated that ectopic expression of hTERT could result in immortalization of FKSCs. Tumorigenecity of hTERT-EGFP-FKSCs were examined by soft agar assay and transplantation into NOD-SCID mice. Results showed that hTERT-EGFP-FKSCs sustained the cellular characteristics of native FKSCs and had no transforming activity. In vivo differentiation multipotentials of hTERT-EGFP-FKSCs were confirmed by transplantation into developing chicken embryos and in situ hybridization analysis. These data provide a novel framework for understanding human telomerase activity in different species and suggest a new insight for manipulating hTERT for therapeutic purposes in treating tissue injury and aging. PMID- 19551766 TI - Hypersensitivity to RF fields emitted from CDMA cellular phones: a provocation study. AB - With the number of cellular phone users rapidly increasing, there is a considerable amount of public concern regarding the effects that electromagnetic fields (EMFs) from cellular phones have on health. People with self-attributed electromagnetic hypersensitivity (EHS) complain of subjective symptoms such as headaches, insomnia, and memory loss, and attribute these symptoms to radio frequency (RF) radiation from cellular phones and/or base stations. However, EHS is difficult to diagnose because it relies on a person's subjective judgment. Various provocation studies have been conducted on EHS caused by Global System for Mobile Communications (GSM) phones in which heart rate and blood pressure or subjective symptoms were investigated. However, there have been few sham controlled provocation studies on EHS with Code Division Multiple Access (CDMA) phones where physiological parameters, subjective symptoms, and perception of RF radiation for EHS and non-EHS groups were simultaneously investigated. In this study, two volunteer groups of 18 self-reported EHS and 19 non-EHS persons were tested for both sham and real RF exposure from CDMA cellular phones with a 300 mW maximum exposure that lasted half an hour. We investigated not only the physiological parameters such as heart rate, respiration rate, and heart rate variability (HRV), but also various subjective symptoms and the perception of EMF. In conclusion, RF exposure did not have any effects on physiological parameters or subjective symptoms in either group. As for EMF perception, there was no evidence that the EHS group better perceived EMF than the non-EHS group. PMID- 19551767 TI - Melatonin as a potential therapeutic agent in psychiatric illness. AB - The aim of this review was to summarize the potential use of melatonin in the treatment of mental disorders, specifically bipolar disorders, depression, and schizophrenia. To date, melatonin has been most commonly used in psychiatry because of its hypnotic, rhythm resynchronizing, and antioxidant actions. Here, we examine other properties of the melatonin including its anti-inflammatory, antinociceptive, anxiolytic, and drug detoxification actions as well as its protective effects against neural loss. The brain is an intricate sensory and motor organ which receives information from both the external and internal environments. It transduces information into complex chemical and electrical signals which are transmitted throughout the central nervous system (CNS) and the organism. The pathogenesis of mental disorders remains ambiguous and neuroinflammation has been proposed as a causative agent. We consider the potential contributions of melatonin as therapeutic agent in CNS and during neuroinflammation in mental disorders. PMID- 19551769 TI - From azides to nitrogen-centered radicals: applications of azide radical chemistry to organic synthesis. AB - Over the last thirty years organic azides have drawn a great deal of attention as radical traps for carbon- and heteroatom-centered radicals, both in intra- and in intermolecular processes. The resulting intermediates (nitrogen-centered radicals such as triazenyls, aminyls, or even iminyls) can be conveniently employed in the synthesis of a variety of cyclic and acyclic nitrogen-containing compounds. PMID- 19551768 TI - The influence of pulsed magnetic fields (PMFs) on nonsynaptic potentials recorded from the central and peripheral nervous systems in vitro. AB - The influence of pulsed magnetic fields (PMFs) on nonsynaptic potentials recorded from the central and peripheral nervous system in vitro has been investigated. The population spikes (PSs) recorded from hippocampal slices during antidromic stimulation and compound action potentials (CAPs) recorded from the segments of the sciatic nerve were used as indicators of neuronal activity. The potentials recorded from both preparations were significantly and permanently enhanced following PMF (0.16 Hz, 15 mT) exposure. The increase in the antidromic PS occurred even in the presence of potassium channel blocker tetraethylammonium (TEA) and was accompanied by multiple spiking. Among all frequencies of PMF tested (0.5, 0.16, 0.07, 0.03, 0.0 Hz), the frequency of 0.5 Hz was the most effective in enhancement of potential amplitude. The influence of PMF on the amplitude of two CAPs evoked by the pair of electrical stimuli applied in rapid succession has also been evaluated. In control conditions the potential triggered by the second stimuli was slightly smaller expressing the phenomenon of short term depression (STD). Although PMF exposure amplified the amplitude of both potentials, the increase in the size of the first potential was significantly greater increasing further the magnitude of STD. The blocking of potassium channels reversed STD into facilitation. One of the possible mechanisms involved in PMF action could be the modification of the axonal threshold, which was significantly reduced following exposure to PMF. PMID- 19551770 TI - The catalytic mechanism of fluoroacetate dehalogenase: a computational exploration of biological dehalogenation. AB - The biological dehalogenation of fluoroacetate carried out by fluoroacetate dehalogenase is discussed by using quantum mechanical/molecular mechanical (QM/MM) calculations for a whole-enzyme model of 10 800 atoms. Substrate fluoroacetate is anchored by a hydrogen-bonding network with water molecules and the surrounding amino acid residues of Arg105, Arg108, His149, Trp150, and Tyr212 in the active site in a similar way to haloalkane dehalogenase. Asp104 is likely to act as a nucleophile to attack the alpha-carbon of fluoroacetate, resulting in the formation of an ester intermediate, which is subsequently hydrolyzed by the nucleophilic attack of a water molecule to the carbonyl carbon atom. The cleavage of the strong C-F bond is greatly facilitated by the hydrogen-bonding interactions between the leaving fluorine atom and the three amino acid residues of His149, Trp150, and Tyr212. The hydrolysis of the ester intermediate is initiated by a proton transfer from the water molecule to His271 and by the simultaneous nucleophilic attack of the water molecule. The transition state and produced tetrahedral intermediate are stabilized by Asp128 and the oxyanion hole composed of Phe34 and Arg105. PMID- 19551771 TI - A mechanistic study of the effects of antioxidants on the formation of malondialdehyde-like products in the reaction of hydroxyl radicals with deoxyribose. AB - The reactions of (*)OH radicals with deoxyribose, DR, form five different DR(*) radicals, only one of which is transformed into malondialdehyde (MDA)-like products. The radiolytic yield of the MDA-like products increases with the increase in the DR concentration indicating that some of the initially formed "unproductive" radicals react with DR to form the "productive" radicals. The yield of the MDA-like products also increases with the dose rate delivered to the solution suggesting that the formation of the MDA-like products involves the reaction of the "productive" radicals with a radical. The addition of ascorbate, AH(-), to the solution decreases the yield of the MDA-like products as expected from the relative rates of the reaction of DR and AH(-) with (*)OH radicals. On the other hand the addition of the exogenous thiol, N-acetylcysteine (NAC), to the solutions decreases the yield of the MDA-like products considerably more than expected from the rate constants of the reaction with (*)OH radicals. The addition of the endogenous thiol, glutathione (GSH), to the solutions affects the yield of the MDA-like products at low concentration less than expected and at "high" concentrations more than expected from the rate constant of the reaction. Addition of low concentration of AH(-) to solutions containing GSH increases considerably its antioxidant activity whereas addition of small concentrations of AH(-) to solutions containing NAC has no effect on its antioxidant activity. The results point out that the DR(*) radicals react differently with NAC and GSH and that the GS(*) and NAC(*) radicals react differently with DR, the GS(*) radical being considerably more active than the NAC(*) radical. Thus it has to be concluded that the relative activity of antioxidants depends also on the rate constants of many secondary reactions and on the concentrations of all the solutes present in the system. PMID- 19551772 TI - Reversible uptake of COS, CS2, and SO2: ionic liquids with O-alkylxanthate, O alkylthiocarbonyl, and O-alkylsulfite anions. AB - CO(2)-binding organic liquids (CO(2)BOLs) are mixtures of a base (typically an amidine or guanidine) and an alcohol, and have been shown to reversibly capture and release CO(2) with low reaction energies and high gravimetric CO(2) capacity. We now report the ability of such liquid blends to chemically bind and release other acid gases such as CS(2), COS, and SO(2) analogously to CO(2). These systems bind with sulfur-containing acid gases to form colored ionic liquids with new O-alkylxanthate, O-alkylthiocarbonyl, and O-alkylsulfite anions. The capture and thermal stripping of each acid gas from these systems and their applicability towards flue gas desulfurization is discussed. PMID- 19551773 TI - Framework breathing in the vapour-phase adsorption and separation of xylene isomers with the metal-organic framework MIL-53. AB - Vapour-phase adsorption and separation of the C8 alkyl aromatic compounds p xylene, m-xylene, o-xylene, and ethylbenzene has been studied on the metal organic framework MIL-53. Adsorption and desorption isotherms of the pure components at 110 degrees C were determined using the gravimetric technique. The adsorption isotherms show two well-defined steps and hysteresis, corresponding to the opening or breathing of the framework, as induced by the presence of the adsorbing molecules. In the first isotherm plateau, an adsorption capacity of about 18 wt % is observed. After the breathing phenomenon, the adsorption capacity increases to about 40 wt %. Breakthrough separation experiments with equimolar o-xylene/ethylbenzene mixtures were performed at 110 degrees C with varying hydrocarbon pressures. The separation mechanism is related to the state of the pore structure, as dictated by framework breathing. At low pressure, below the "pore-opening" pressure, MIL-53 shows no preference for any isomer. At pressures high enough to induce pore opening, separation of the C8 alkyl aromatic isomers becomes possible and separation factors as high as 6.5 are observed. The separation at a high degree of pore filling in the open form is a result of differences in the packing modes of the C8 alkyl aromatic components in the pores of MIL-53. PMID- 19551774 TI - Heterogeneous wheel-shaped Cu20-polyoxotungstate [Cu20Cl(OH)24(H2O)12(P8W48O184)]25- catalyst for solvent-free aerobic oxidation of n-hexadecane. AB - The selective oxidation of alkanes as a green process remains a challenging task because partial oxidation is easier to achieve with sacrificial oxidants, such as hydrogen peroxide, alkyl hydroperoxides or iodosylbenzene, than with molecular oxygen or air. Here, we report on a heterogeneous catalyst for n-hexadecane oxidation comprised of the wheel shaped Cu20-polyoxotungstate [Cu20Cl(OH)24(H2O)12(P8W48O184)]25- anchored on 3-aminopropyltriethoxysilane (apts)-modified SBA-15. The catalysts were characterized by powder X-ray diffraction (XRD), N2-adsorption measurements and Fourier transform infrared reflectance (FT-IR) spectroscopy. The heterogeneous Cu20-polyanion system catalyzed the solvent-free aerobic oxidation of n-hexadecane to alcohols and ketones by using air as the oxidant under ambient conditions. The catalyst exhibits an exceptionally high turn over frequency (TOF) of 20,000 h(-1) at 150 degrees C and is resistant to poisoning by CS2. Moreover, it can be easily recovered and reused by filtration without loss of its catalytic activity. Possible homogeneous contributions also have been examined and eliminated. Thus, this system can use air as oxidant, which, in combination with its good overall performance and poison tolerance, raises the prospect of this type of heterogeneous catalyst for practical applications. PMID- 19551775 TI - Modelling the magnetic behaviour of square-pyramidal Co(II)5 aggregates: tuning SMM behaviour through variations in the ligand shell. AB - Three new mu4-bridged Co(II)5 clusters with similar core motifs have been synthesised with the use of N-tert-butyldiethanolamine (tbdeaH2) and pivalic acid (piv): [Co(II)5(mu4-N3)(tbdea)2(mu-piv)4(piv)(CH3CN)2].CH3CN (1), [Co(II)5(mu4 Cl)(Cl)(tbdea)2(mu-piv)4(pivH)2] (2) and [Co(II)5(mu4-N3)(Cl)(tbdea)2(mu piv)4(pivH)2] (3). Magnetic measurements were performed for all three compounds. It was found that while the chloride-bridged cluster 2 does not show an out-of phase signal, which excludes single-molecule magnet (SMM) behaviour, the azide bridged compounds 1 and 3 show out-of-phase signals as well as frequency dependence of the ac susceptibility, as expected for SMMs. We confirmed that 1 is a SMM with zero-field quantum tunnelling of the magnetisation at 1.8 K. Compound 3 is likely a SMM with a blocking temperature well below 1.8 K. We established a physical model to fit the chiT versus T and M versus B curves of the three compounds to reproduce the observed SMM trend. The analysis showed that small changes in the ligand shell modify not only the magnitude of exchange constants, but also affect the J and g matrices in a non-trivial way. PMID- 19551777 TI - Synthesis of amphiphilic [60]fullerene [3:3]hexakisadducts with four spherically defined addend zones. AB - New [3:3]hexakisadducts 3, 6, and 7 have been synthesized by a stepwise addition of two tripodal malonate tethers to pristine [60]fullerene. For the first time, [3:3]hexakisadducts of two sets of asymmetrically substituted malonate addends could be prepared as single in/out isomers by following this synthetic protocol. Thereby, four spherically defined addend zones I-IV were created with various similar degrees of latitude on the fullerene sphere. Four amphiphilic [3:3]hexakisadducts 12, 14, 18, and 19 were prepared with different relative arrangements of both the hydrophilic and lipophilic moieties in the addend zones I/II and III/IV, respectively. PMID- 19551776 TI - Direct imidation to construct 1H-benzo[d]imidazole through Pd(II)-catalyzed C-H activation promoted by thiourea. PMID- 19551778 TI - High-throughput reaction optimisation and activity screening of ferrocene-based Lewis acid-catalyst complexes by using continuous-flow reaction detection mass spectrometry. AB - Optimising synthetic conversions and assessing catalyst performance is a tedious and laborious endeavour. Herein, we present an automated alternative to the commonly applied sequential approaches that are used to increase catalyst discovery process efficiencies by increasing the number of entities that can be tested. This new approach combines conversion of the reactants and determination of product formation into a single comprehensive reaction detection system that can be operated with minimal catalyst and reactant consumption. With this approach, rudimentary reaction conditions can be quickly optimised and the same system can then be used to screen for the optimal homogenous catalyst in a selected solution-phase synthetic conversion. The system, which is composed of standard HPLC components, can be used to screen catalyst libraries at a repetition rate of five minutes and can be run unsupervised. The sensitive mass spectrometric detection that is implemented in the reaction detection methodology can be used for the simultaneous monitoring of reactants, catalysts and product ions. In the experiments, the three-component reaction that gives a substituted 2 imidazoline was optimised. Afterwards, the same method was used to assess a library of ferrocene-based Lewis acid catalysts for performance in the aforementioned conversion in six different solvents. We demonstrate the feasibility of using this methodology to directly compare the performance results obtained in different solvents by calibrating the solvent-specific MS responses. PMID- 19551779 TI - Large di- and heptafullerene polyelectrolytes composed of C60 building blocks having a highly symmetrical hexakisaddition pattern. AB - We report here on the synthesis of three new prototypes (types I-III) of very large fullerene-based polyelectrolytes which can carry up to 60 charges on their periphery. All fullerene moieties incorporated in these macromolecular structures have an octahedral hexakisaddition pattern. Dumbbell-shaped icosacarboxylate 5 (type I), which can accumulate up to twenty negative charges, is very soluble in methanol as well as in neutral and basic water. On the other hand, Janus dumbbell 13 (type II) contains both positively and negatively chargeable fullerene building blocks and is very soluble in acidic and basic media. However, in the region of the isoelectric point at pH 6.0-6.5 it precipitates as a pale orange solid due to pronounced intermolecular Coulomb interactions. Giant heptafullerene 15 (type III) can store up to 60 positive charges in its periphery and is the largest molecular polyelectrolyte with defined three-dimensional structure. PMID- 19551780 TI - Synthesis and orthogonal functionalization of [60]fullerene e,e,e-trisadducts with two spherically defined addend zones. AB - e,e,e-Trisadducts 13 and 15 have been prepared by a highly regioselective threefold cyclopropanation of tripodal malonates 10 and 12 with C60. The yield and regioselectivity depend on the length and structure of the tethers that connect the malonate units to the focal benzene core of 13-15. As a consequence of the template-directed synthesis, all e,e,e-trisadducts were formed as in/out isomers exclusively and contain two spherically well-defined addend zones with equatorial and polar orientation, respectively. By variation of the outer malonate termini of the tethers, selective functionalization of the equatorial addend zone could be achieved, thus leading to fine-tuning of intermolecular interactions, such as solubility or aggregation phenomena. After removal of the focal benzene moiety in 14 and 15, selective functionalization of the polar addend zone could be achieved. Strong intramolecular hydrogen-bonding networks of the polar substituents in the polar addend zone could be observed by 1H NMR spectroscopic analysis. By orthogonal functionalization of both addend zones, fullerene derivatives 44-48 could be synthesized as one single in/out isomer, thus greatly enhancing the potential of e,e,e-trisadducts as building blocks in supramolecular architectures. PMID- 19551781 TI - Polydiacetylene-based colorimetric self-assembled vesicular receptors for biological phosphate ion recognition. AB - Self-assembled vesicular polydiacetylene (PDA) particles with embedded metal complex receptor sites have been prepared. The particles respond to the presence of ATP and PPi (pyrophosphate) in buffered aqueous solution by visible changes of their color and emission properties. Blue PDA vesicles of uniform size of about 200 nm were obtained upon UV irradiation from mono- and dinuclear zinc(II)-cyclen and iminodiacetato copper [Cu(II)-IDA] modified diacetylenes, embedded in amphiphilic diacetylene monomers. Addition of ATP and PPi to the PDA vesicle solution induces a color change from blue to red observable by the naked eye. The binding of ATP and PPi changes the emission intensity. Other anions such as ADP, AMP, H2PO4-, CH3COO-, F-, Cl-, Br- and I-, failed to induce any spectral changes. The zinc(II)-cyclen nanoparticles are useful for the facile detection of PPi and ATP in millimolar concentrations in neutral aqueous solutions, while Cu(II)-IDA modified vesicular PDA receptors are able to selectively discriminate between ATP and PPi. PMID- 19551782 TI - Selective triplet-state formation during charge recombination in a fullerene/Bodipy molecular dyad (Bodipy=borondipyrromethene). AB - A conformationally restricted molecular dyad has been synthesized and subjected to detailed photophysical examination. The dyad comprises a borondipyrromethene (Bodipy) dye covalently linked to a buckminsterfullerene C60 residue, and is equipped with hexadecyne units at the boron centre in order to assist solubility. The linkage consists of a diphenyltolane, attached at the meso position of the Bodipy core and through an N-methylpyrrolidine ring at the C60 surface. Triplet states localised on the two terminals are essentially isoenergetic. Cyclic voltammetry indicates that light-induced electron transfer from Bodipy to C60 is thermodynamically favourable and could compete with intramolecular energy transfer in the same direction. The driving force for light-induced electron abstraction from Bodipy by the singlet excited state of C60 depends critically on the solvent polarity. Thus, in non-polar solvents, light-induced electron transfer is thermodynamically uphill, but fast excitation energy transfer occurs from Bodipy to C60 and is followed by intersystem crossing and subsequent equilibration of the two triplet excited states. Moving to a polar solvent switches on light-induced electron transfer. Now, in benzonitrile, the charge transfer state (CTS) is positioned slightly below the triplet levels, such that charge recombination restores the ground state. However, in CH2Cl2 or methyltetrahydrofuran, the CTS is slightly higher in energy than the triplet levels, and decays, in part, to form the triplet state localized on the C60 residue. This step is highly specific and does not result in direct formation of the triplet excited state localized on the Bodipy unit. Subsequent equilibration of the two triplets takes place on a relatively slow timescale. PMID- 19551784 TI - Stereochemistry of compounds with coordination number ten. AB - The stereochemistry of ten-coordinate rare-earth and transition-metal compounds is studied from the point of view of continuous shape measures (CShM) and derived tools. A total of 19 ideal ten-vertex polyhedra belonging to 12 different symmetry point groups have been considered, from which nine are retained for the description of the stereochemistries of all studied compounds. The structures of the coordination spheres are analyzed by families, according to the denticity and topology of the ligands. Some ligand topologies are seen to consistently favor usually neglected ideal polyhedra, such as the capped trigonal cupola. PMID- 19551783 TI - Solvent-enhanced diastereo- and regioselectivity in the Pd(II)-catalyzed synthesis of six- and eight-membered heterocycles via cis-aminopalladation. AB - The Pd(II)-catalyzed intramolecular oxidative cyclization of tosyl-protected cis- and trans-N-allyl-2-aminocyclohexanecarboxamides was examined, and efficient syntheses of cyclohexane-fused pyrimidin-4-ones and 1,5-diazocin-6-ones were developed. In the course of the research, a marked solvent effect was observed on both the regio- and diastereoselectivity. Additionally, a novel Pd(II)-mediated domino oxidation, oxidative amination reaction was discovered. Our experimental and theoretical findings suggest that the reactions proceed via a cis aminopalladation mechanism. PMID- 19551785 TI - New blue-light-emitting ultralong [Cd(L)(TeO3)] (L = polyamine) organic-inorganic hybrid nanofibre bundles: their thermal stability and acidic sensitivity. AB - A new type of blue-light-emitting ultralong [Cd(L)(TeO(3))] (L = ethylenediamine, diethylenetriamine) nanofibre bundle has been synthesised under reflux in a mixed solvent media. Inorganic Cd(TeO(3)) layers are assumed to exist in the structures and are connected by the organic amine molecules through the coordination between nitrogen atoms and cadmium ions. The composition and formulae of these hybrid materials, based on the proposed structures, have been identified through element analysis (EA), thermal gravity analysis (TGA) and energy dispersive spectra (EDS). The thermal stabilities and optical properties of these nanofibre bundles have been investigated. Thermal decomposition of [Cd(en)(TeO(3))] (en = ethylenediamine) and [Cd(DETA)(TeO(3))] (DETA = diethylenetriamine) at 450 degrees C allowed the formation of a mixture of CdTe and Cd(TeO(3)) phases, and a pure CdTe phase, respectively. In addition, this new kind of hybrid bundle, which demonstrates blue emission, was found to be sensitive to acids, and the emission intensity is strongly dependent on the acidity of the solutions, implying that these hybrid nanofibre bundles could be potentially applied as acid sensors. PMID- 19551786 TI - Structural adaptability of zinc binding sites: different structures in partially, fully, and heavy-metal loaded states. AB - The present study demonstrates that both the nature (Zn(II), Cd(II) or Hg(II)) and supply of metal ions determine whether zinc fingers fold into the well-known, fully loaded structures or alternatively populate a variety of structural states under substoichiometric conditions. Metal-bridged species are observed by perturbed angular correlation (PAC), EXAFS, UV spectroscopy, and stopped-flow kinetics. Transitions between structural states as adaptive reactions to changed metal-ion supply might represent intelligent system changes in zinc homeostasis, trafficking and signalling, and reflect features of heavy-metal toxicity at the molecular level. Because the zinc fingers exist in structural states that are different from the metal-free and fully loaded species, the prevailing view on metal-mediated molecular regulation in terms of "on and off control" might be oversimplified. PMID- 19551788 TI - Do extremely bent allenes exist? PMID- 19551787 TI - Electron-transfer kinetics of microperoxidase-11 covalently immobilised onto the surface of multi-walled carbon nanotubes by reactive landing of mass-selected ions. AB - Controlled deposition of biological molecules on nanostructured materials is a basic step towards the realisation of biochip components. In this study we report the investigation of the first covalent immobilisation of mass-selected redox protein on a carboxyl-functionalised multi-walled carbon nanotube (MWCNT) electrode surface by means of ion soft landing. The immobilised protein maintains its biochemical properties, displaying an excellent electrochemical behaviour on the electrode surface. The deposition of mass-selected ions is influenced by several factors, including the charge state and the collision energy of the projectile ions. To elucidate the mechanism involved in the protein reactive landing onto the MWCNT surface, the data obtained from cyclic voltammetry experiments were modelled according to the Marcus theory. The proposed method opens up the way to the development of a new generation of biocomponents with potential use in biosensors, diagnostics, biofuel cells and bioactive films. PMID- 19551789 TI - N-heteroacenes. AB - The synthesis and the property evaluation of several large N-heteroacenes are discussed. Issues of stability and aromaticity are compared and investigated and a historical perspective of the field is given. Some of the larger heterocyclic materials that are evaluated in this concept article have been around for more than one hundred years, yet their chemistry and properties are not well known/understood. PMID- 19551790 TI - Synthesis and structure-activity correlation studies of secondary- and tertiary amine-based glutathione peroxidase mimics. AB - In this study, a series of secondary- and tertiary-amino-substituted diaryl diselenides were synthesized and studied for their glutathione peroxidase (GPx) like antioxidant activities with H(2)O(2), cumene hydroperoxide, or tBuOOH as substrates and with PhSH or glutathione (GSH) as thiol cosubstrates. This study reveals that replacement of the tert-amino groups in benzylamine-based diselenides by sec-amino moieties drastically enhances the catalytic activities in both the aromatic thiol (PhSH) and GSH assay systems. Particularly, the N propyl- and N-isopropylamino-substituted diselenides are 8-18 times more active than the corresponding N,N-dipropyl- and N,N-diisopropylamine-based compounds in all three peroxide systems when GSH is used as the thiol cosubstrate. Although the catalytic mechanism of sec-amino-substituted diselenides is similar to that of the tert-amine-based compounds, differences in the stability and reactivity of some of the key intermediates account for the differences in the GPx-like activities. It is observed that the sec-amino groups are better than the tert amino moieties for generating the catalytically active selenols. This is due to the absence of any significant thiol-exchange reactions in the selenenyl sulfides derived from sec-amine-based diselenides. Furthermore, the seleninic acids (RSeO(2)H) derived from the sec-amine-based compounds are more stable toward further reactions with peroxides than their tert-amine-based analogues. PMID- 19551791 TI - Stabilization of the acyclic tautomer in reducing carbohydrates. PMID- 19551792 TI - Formal highly enantioselective organocatalytic addition of fluoromethyl anion to alpha,beta-unsaturated aldehydes. PMID- 19551793 TI - Lewis base stabilized dichlorosilylene. PMID- 19551794 TI - Synthesis and structure of stereoisomeric multivicinal hexafluoroalkanes. PMID- 19551795 TI - Eosin Y-sensitized artificial photosynthesis by highly efficient visible-light driven regeneration of nicotinamide cofactor. AB - Dye-sensitized photosynthesis: Eosin Y (EY), a dye photosensitizer, works efficiently as a molecular photoelectrode by catalyzing the visible-light-driven electron-transfer reaction for efficient regeneration of NADH through a photosensitizer-electron relay dyad. Injection of the photosensitized electron resulted in highly accelerated regeneration of NADH, which can be used by glutamate dehydrogenase for the photosynthesis of L-glutamate. PMID- 19551796 TI - Translational diffusion and interaction of a photoreceptor and its cognate transducer observed in giant unilamellar vesicles by using dual-focus FCS. AB - In order to monitor membrane-protein binding in lipid bilayers at physiological protein concentrations, we employed the recently developed dual-focus fluorescence correlation spectroscopy (2fFCS) technique. In a case study on a photoreceptor consisting of seven transmembrane helices and its cognate transducer (two transmembrane helices), the lateral diffusion for these integral membrane proteins was analyzed in giant unilamellar vesicles (GUVs). The two dimensional diffusion coefficients of both separately diffusing proteins differ significantly, with D = 2.2 x 10(-8) cm2 s(-1) for the photoreceptor and with D = 4.1 x 10(-8) cm2 s(-1) for the transducer. In GUVs with both membrane proteins present together, we observed significantly smaller diffusion coefficients for labelled transducer molecules; this indicates the presence of larger diffusing units and therefore intermolecular protein binding. Based on the phenomenological dependence of diffusion coefficients on the molecule's cylindrical radius, we are able to estimate the degree of membrane protein binding on a quantitative level. PMID- 19551797 TI - Spatio-temporal control of cell coculture interactions on surfaces. AB - Coculture control: We report a combined photochemical and electroactive self assembled monolayer (SAM)-based substrate strategy to generate a coculture platform with spatial and temporal control of cell-cell interactions. These dynamic substrates can present a variety of ligands on the surface for biospecific interactions between the ligands and cell surface receptors. Photopatterning enables the ligands to be immobilized in patterns and even gradients. PMID- 19551798 TI - Magnetoelectric memories--a disruptive technology? PMID- 19551799 TI - Synthesis and antimalarial properties of streptocyanine dyes. AB - Several streptocyanine dyes were synthesized that contain polymethine chains of varying length. Their in vitro antimalarial activities were evaluated against the virulent P. falciparum parasite. In addition to the influence of polymethine chain length, the effects of structural modifications at nitrogen end groups, para substitution of the phenyl groups, and counter-anions were studied. The most potent antimalarial activities were found for heptacarbon chain streptocyanines, with an IC(50) value of 60 nM. Interestingly, most of the compounds were less cytotoxic toward the mammalian cells tested. The best selective toxicity profiles were found for pentacarbon chain streptocyanines, which have a good in vitro specificity index. PMID- 19551800 TI - Screening for the drug-phospholipid interaction: correlation to phospholipidosis. AB - Phospholipid bilayers represent a complex, anisotropic environment fundamentally different from bulk oil or octanol, for instance. Even "simple" drug association to phospholipid bilayers can only be fully understood if the slab-of-hydrocarbon approach is abandoned and the complex, anisotropic properties of lipid bilayers reflecting the chemical structures and organization of the constituent phospholipids are considered. The interactions of drugs with phospholipids are important in various processes, such as drug absorption, tissue distribution, and subcellular distribution. In addition, drug-lipid interactions may lead to changes in lipid-dependent protein activities, and further, to functional and morphological changes in cells, a prominent example being the phospholipidosis (PLD) induced by cationic amphiphilic drugs. Herein we briefly review drug-lipid interactions in general and the significance of these interactions in PLD in particular. We also focus on a potential causal connection between drug-induced PLD and steatohepatitis, which is induced by some cationic amphiphilic drugs. PMID- 19551801 TI - Stereoselective HDAC inhibition from cysteine-derived zinc-binding groups. AB - A series of small-molecule histone deacetylase (HDAC) inhibitors, which feature zinc binding groups derived from cysteine, were synthesized. These inhibitors were tested against multiple HDAC isoforms, and the most potent, compound 10, was determined to have IC(50) values below 1 microM. The compounds were also tested in a cellular assay of oxidative stress-induced neurodegeneration. Many of the inhibitors gave near-complete protection against cell death at 10 microM without the neurotoxicity seen with hydroxamic acid-based inhibitors, and were far more neuroprotective than HDAC inhibitors currently in clinical trials. Both enantiomers of cysteine were used in the synthesis of a variety of novel zinc binding groups (ZBGs). Derivatives of L-cysteine were active in the HDAC inhibition assays, while the derivatives of D-cysteine were inactive. Notably, the finding that both the D- and L-cysteine derivatives were active in the neuroprotection assays suggests that multiple mechanisms are working to protect the neurons from cell death. Molecular modeling was employed to investigate the differences in inhibitory activity between the HDAC inhibitors generated from the two enantiomeric forms of cysteine. PMID- 19551802 TI - Design, selection, and evaluation of a general kinase-focused library. PMID- 19551803 TI - Theoretical screening of ionic liquid solvents for carbon capture. PMID- 19551804 TI - Vanadium-catalyzed oxidative bromination under atmospheric oxygen. PMID- 19551807 TI - Successful weaning after plasma exchange for polyneuropathy related to POEMS syndrome. AB - Respiratory failure due to polyneuropathy associated with POEMS syndrome (Polyneuropathy, organomegaly, endocrinopathy, M protein, and skin changes) is unusual. In this report, we describe a 47-year-old male patient with polyneuropathy related to POEMS syndrome who suffered from hypercapnic respiratory failure due to progressive weakness of the respiratory muscles. His respiratory muscle strength significantly improved after plasma exchange and was extubated successfully. To our knowledge, this is the first case of POEMS syndrome with aggressive polyneuropathy that was successful treated by plasma exchange and our experience provides insight regarding the optimal management of such patients. PMID- 19551805 TI - Ginkgo biloba: specificity of neuropsychological improvement--a selective review in search of differential effects. AB - Extracts of Ginkgo biloba are widely used for the treatment of cognitive impairment. Whereas reviews have focused on the question whether ginkgo is effective to enhance cognition in general, little is known about specificity of improvement. This might be crucial for future trials, thus enabling hypotheses about sensitive outcome measures. Therefore, this article summarizes such information, i.e. neuropsychological effects of chronic administration of ginkgo in healthy and cognitively impaired subjects of any age. Objective psychometric test results were considered if they reflected distinct cognitive functions from randomized controlled group-studies (RCT). We reviewed 29 RCTs yielding 209 placebo-drug comparisons of psychometric scores in four different cognitive domains comprising 14 sub-functions. Whereas little specific information can be obtained from trials for treatment of dementia, a pattern of pharmacological actions on cognitive processes emerges here from studies for mild cognitive impairment (MCI), depression, multiple sclerosis and healthy young and elderly subjects. There is consistent evidence that chronic administration improves selective attention, some executive processes and long-term memory for verbal and non-verbal material. Further trials should be more comprehensive as there are few data available on some cognitive functions and psychometric flaws in the selection of tests and the interpretation of their results favouring predominantly beta-errors. Thus, though this pattern is encouraging it also asks for a cautious interpretation to date. PMID- 19551808 TI - Passage marker excretion in red kangaroo (Macropus rufus), collared peccary (Pecari tajacu) and colobine monkeys (Colobus angolensis, C. polykomos, Trachypithecus johnii). AB - Ruminants are characterized by an efficient particle-sorting mechanism in the forestomach (FRST) followed by selective rechewing of large food particles. For the nonruminating foregut fermenter pygmy hippo it was demonstrated that large particles are excreted as fast as, or faster than, the small particles. The same has been suggested for other nonruminating foregut fermenters. We determined the mean retention time of fluids and different-sized particles in six red kangaroos (Macropus rufus), seven collared peccaries (Pecari tajacu) and three colobine monkeys (Colobus angolensis, C. polykomos, Trachypithecus johnii). We fed Co-EDTA as fluid and mordanted fiber as particle markers (Cr, Ce). Mean (+ or - SD) total tract retention time for fluids, small and large particles was 14 + or - 2, 29 + or - 10 and 30 + or - 9 hr in red kangaroos, 26 + or - 2, 34 + or - 5 and 32 + or - 3 hr in collared peccaries and 57 + or - 17, 55 + or - 19 and 54 + or - 19 hr in colobine monkeys, respectively. Large and small particles were excreted simultaneously in all species. There was no difference in the excretion of fluids and particles in the colobine monkeys, in contrast to the other foregut fermenters. In the nonprimate, nonruminant foregut fermenters, the difference in the excretion of fluids and small particles decreases with increasing food intake. On the contrary, ruminants keep this differential excretion constant at different intake levels. This may be a prerequisite for the sorting of particles in their FRST and enable them to achieve higher food intake rates. The functional significance of differential excretion of fluids and particles from the FRST requires further investigations. PMID- 19551809 TI - Upregulation of plakophilin-2 and its acquisition to adherens junctions identifies a novel molecular ensemble of cell-cell-attachment characteristic for transformed mesenchymal cells. AB - In contrast to the desmosome-containing epithelial and carcinoma cells, normal and malignantly transformed cells derived from mesenchymal tissues and tumors are connected only by adherens junctions (AJs) containing N-cadherins and/or cadherin 11, anchored in a cytoplasmic plaque assembled by alpha- and beta-catenin, plakoglobin, proteins p120 and p0071. Here, we report that the AJs of many malignantly transformed cell lines are characterized by the additional presence of plakophilin-2 (Pkp2), a protein hitherto known only as a major component of desmosomal plaques, i.e., AJs of epithelia and carcinomatous cells. This massive acquisition of Pkp2 and its integration into AJ plaques of a large number of transformed cell lines is demonstrated with biochemical and immunolocalization techniques. Upregulation of Pkp2 and its integration into AJs has also been noted in some soft tissue tumors insitu and some highly proliferative colonies of cultured mesenchymal stem cells. As Pkp2 has recently been identified as a functionally important major regulatory organizer in AJs and related junctions in epithelial cells and cardiomyocytes, we hypothesize that the integration of Pkp2 into AJs of "soft tissue tumor" cells also can serve functions in the upregulation of proliferation, the promotion of malignant growth in general as well as the close-packing of diverse kinds of cells and the metastatic behavior of such tumors. We propose to examine its presence in transformed mesenchymal cells and related tumors and to use it as an additional diagnostic criterion. PMID- 19551810 TI - A comparison of metabolite extraction strategies for 1H-NMR-based metabolic profiling using mature leaf tissue from the model plant Arabidopsis thaliana. AB - Metabolite analysis is recognized as an important facet of systems biology, however complete metabolome characterization has not been realized due to challenges in sample preparation, inherent instrumental limitations and the labor intensive task of data interpretation. This work aims to compare several commonly used metabolite extraction strategies for their effect on the (1)H nuclear magnetic resonance (NMR) metabolic profile of extracts of the model plant Arabidopsis thaliana. Extractions were carried out on aliquots from a pool of homogenized plant tissue using CD(3)CN/D(2)O, buffered D(2)O, perchloric acid in D(2)O, CD(3)OD/D(2)O and CD(3)OD/D(2)O/CDCl(3) as the extraction solvents. The effects of lyophilization as a sample pretreatment, solvent evaporation and extract fractionation for removal of interfering species were studied. Representative spectra are presented for qualitative interpretation. Analytical reproducibility was evaluated by principal components analysis. Perchloric acid facilitated acid-catalyzed cleavage of sucrose, further complicating biological interpretation of the resulting metabolite profile. The solvent system CD(3)OD/D(2)O/CDCl(3) gave the least reproducible results in our hands. D(2)O extracts suffered from poor stability probably due to contamination by soluble enzymes, which were not denatured in this solvent. CD(3)CN/D(2)O extracts showed greater stability than D(2)O alone, but problems were encountered due to degradation of (1)H NMR spectral resolution during lengthy acquisitions due to partial phase separation. In addition, this solvent system produced spectra with significant contamination by lipids that obscured spectral regions containing the resonances of the aliphatic amino acids. These problems were solved by speedvacuuming the CD(3)CN/D(2)O extract and reconstituting in D(2)O solution. PMID- 19551811 TI - Concordance analysis of an in vitro micronucleus screening assay and the regulatory chromosome aberration assay using pharmaceutical drug candidates. AB - The in vitro micronucleus assay is under consideration by regulatory agencies as a suitable alternative to the in vitro chromosome aberration (CA) assay. At Pfizer, we utilized a non-Good Laboratory practices cytokinesis-block in vitro micronucleus (CBMN) assay in CHO cells as a screen to predict the regulatory outcome of the human lymphocyte CA assay, and we have retrospectively analyzed a highly select set of 112 internal drug candidates to measure concordance. Overall, our exploratory CBMN correctly classified 97 of 112 (86.6%) compounds in the CA assay. Specificity was high with 87 of 92 (94.6%) CA negative compounds correctly classified by CBMN. Sensitivity was low at 50% with 10 of 20 CA positive compounds correctly classified by CBMN; this may be attributed to the low number of CA positives in the select set. In an attempt to improve sensitivity, we increased the number of CA positives by combining our internal set with an industrial data set previously published (Miller B et al. 1997: Mutat Res 392:45-59). When combined, concordance was 86.7% (143/165), specificity was 91.2% (114/125), and sensitivity increased to 72.5% (29/40). Because cytotoxicity is considered a confounding factor of in vitro test systems, we also examined, within the Pfizer data set, the influence of cytotoxicity in the CBMN assay, and the results indicated that seemingly low (<50%) or excessively high (>70%) levels of cytotoxicity did not significantly alter predicted CA outcome. These collective analyses contribute to growing evidence that the CBMN assay is a suitable regulatory option in the standard battery of genetic toxicology tests. PMID- 19551812 TI - Analysis of genomic dose-response information on arsenic to inform key events in a mode of action for carcinogenicity. AB - A comprehensive literature search was conducted to identify information on gene expression changes following exposures to inorganic arsenic compounds. This information was organized by compound, exposure, dose/concentration, species, tissue, and cell type. A concentration-related hierarchy of responses was observed, beginning with changes in gene/protein expression associated with adaptive responses (e.g., preinflammatory responses, delay of apoptosis). Between 0.1 and 10 microM, additional gene/protein expression changes related to oxidative stress, proteotoxicity, inflammation, and proliferative signaling occur along with those related to DNA repair, cell cycle G2/M checkpoint control, and induction of apoptosis. At higher concentrations (10-100 microM), changes in apoptotic genes dominate. Comparisons of primary cell results with those obtained from immortalized or tumor-derived cell lines were also evaluated to determine the extent to which similar responses are observed across cell lines. Although immortalized cells appear to respond similarly to primary cells, caution must be exercised in using gene expression data from tumor-derived cell lines, where inactivation or overexpression of key genes (e.g., p53, Bcl-2) may lead to altered genomic responses. Data from acute in vivo exposures are of limited value for evaluating the dose-response for gene expression, because of the transient, variable, and uncertain nature of tissue exposure in these studies. The available in vitro gene expression data, together with information on the metabolism and protein binding of arsenic compounds, provide evidence of a mode of action for inorganic arsenic carcinogenicity involving interactions with critical proteins, such as those involved in DNA repair, overlaid against a background of chemical stress, including proteotoxicity and depletion of nonprotein sulfhydryls. The inhibition of DNA repair under conditions of toxicity and proliferative pressure may compromise the ability of cells to maintain the integrity of their DNA. PMID- 19551813 TI - Aromatic-ring-functionalised benzoxazinones in the system Oryza sativa Echinochloa crus-galli as biorational herbicide models. AB - BACKGROUND: Barnyardgrass, Echinochloa crus-galli (L.) Beauv., is one of the most problematic weeds occurring in rice crops. Although efficient chemical control is provided by herbicides available on the market, resistant biotypes provoked by pressure selection have appeared in recent times. This emphasises the need for alternative treatments in which herbicidal compounds from a natural origin could be included. RESULTS: A number of chemicals with a [2H]-1,4-benzoxazin-3(4H)-one (D-DIBOA) skeleton were tested on this weed, and also in rice, in order to achieve an optimal lead for herbicide composition development by taking into consideration phytotoxic effects and selectivity on the weed. 6-Cl-D-DIBOA causes the same effect as the commercial herbicide propanil at a concentration 15 times lower, while 6-F-D-DIBOA causes this inhibition at a dose 30 times lower. The phytotoxicities caused by 8-Cl-D-DIBOA (IC50 = 44 microM, R2 = 0.866) and 7,8-diF D-DIBOA (IC50 = 52 microM, R2 = 0.9067) are also remarkable. 8-Cl-D-DIBOA was the compound that presented the highest selectivity on Echinochloa crus-galli. The structural requirements for optimal phytotoxicity and selectivity were elucidated by means of QSAR methodology, considering electronic and steric factors. One of the most important descriptors influencing the bioactivity was the dipole moment modulus. This was successfully correlated by employing a second-order polynomial model. CONCLUSION: The in vitro phytotoxic profiles and selectivities shown for these chemicals make them truly promising candidates for higher-level studies. 6F and 6Cl-D-DIBOA, for their high phytotoxicities, and 8-Cl-D-DIBOA, because of its high selectivity, were found to be the most interesting compounds from this point of view. PMID- 19551815 TI - ERK activation is involved in tooth development via FGF10 signaling. AB - The tooth is one of the ectodermal organs that develop from epithelial mesenchymal interactions during embryonic development. An understanding of the underlying molecular mechanisms would improve our knowledge of the growth factors that regulate cell proliferation and differentiation. One of the related aspects is mitogen-activated protein kinase (MAPK) signaling in tooth differentiation. The extracellular-signal regulated kinase (ERK)/mitogen-activated protein kinase kinase (MEK) cascade plays a pivotal role in many of the essential cellular processes underlying embryonic development, including responses to major developmental changes. However, the role of the ERK pathway in molar development is unclear. This study investigated epithelial patterning and tooth growth in the mouse embryo by monitoring ERK and fibroblast growth factor (FGF) signaling. ERK, MEK, and phosphatase and tensin homolog (PTEN) were activated at different levels and locations in the developing tooth at E13.5 to E16.5 and PN2. ERK was activated in the inner dental epithelium and cervical loop, while PTEN was activated in the outer dental epithelium. In addition, only ERK was activated in secretory ameloblast at PN2. To further define the pathways involving FGF and ERK, tooth germs were cultured in the presence of compounds to inhibit MAPK/ERK mediated signaling. Western blot analysis indicated that pERK2 was strongly activated in the tooth germ. Moreover, the activation level of pERK1 was dramatically increased by exogenous FGF10 alone and by combined treatment with FGF10 and U0126. The reported results will improve our understanding of the unique developmental processes of the dental epithelium and tooth growth, and will help to elucidate the fundamental mechanisms of ERK signaling underlying tooth development. PMID- 19551816 TI - The relationship between genotypic sensitivity score and treatment outcomes in late stage HIV disease after supervised HAART. AB - This study investigated the effect of resistance testing quantified through a genotypic sensitivity score (GSS) on virologic, immunologic, and clinical responses among patients with late stage HIV-1 disease receiving supervised highly active antiretroviral therapy (HAART). Newly admitted patients received drug resistance testing (n = 198) and then HAART supervised by residential health care facilities nurses. After initiating a resistance testing-informed HAART regimen, patients were followed for HIV-1 RNA suppression (<50 copies/ml), mean change in CD4(+) T-cells, new AIDS defining category C opportunistic conditions and death. GSS was constructed using the HAART regimen prescribed after resistance testing and data derived from IAS-USA consensus mutations table with modification. Regressions with generalized estimating equations for robust estimation of standard errors and Cox proportional hazards regression estimated independent associations between GSS and treatment responses. After adjusting for adherence, initial log(10) HIV-1 RNA levels, and other covariates, patients with a GSS > or =3 had significantly greater HIV-1 RNA suppression (adjusted odds ratio (AOR) 2.32; 95% CI 1.14, 4.75). HIV-1 RNA levels were lower among patients with > or =95% adherence, but the effect of GSS on viral suppression was not modified by adherence. Self-rated health status, and baseline CD4(+) T-cell counts independently predicted HIV-1 RNA suppression. GSS did not predict mean change in CD4(+) cells/mm(3) (236 vs. 233, P = 0.92), occurrence of new AIDS defining category C conditions or death. These data support resistance testing guided therapy as an independent predictive factor to improve virologic responses in treatment-experienced patients. PMID- 19551814 TI - Do molecular signals from the conceptus influence endometrium decidualization in rodents? AB - A critical period in establishing pregnancy occurs after the onset of implantation but before placental development. Evidence strongly suggests that abnormalities occurring during this period can result in pregnancy termination or in pre-eclampsia; the latter may lead to small-for-gestational-weight offspring that are likely to be unhealthy. Clearly, events occurring in the endometrium during the implantation process are crucial for proper fetal development and for optimal offspring health. In several mammalian species bi-directional communication between the conceptus and endometrium during implantation is required for successful pregnancy. Although different implantation and placentation modes occur in different mammalian species, common aspects of this bi-directional signaling may exist. The molecular signals from the trophoblast cells of the conceptus, which direct endometrial changes during implantation progression, are well known in some nonrodent species. Currently, we know little about such signaling in rodents during implantation progression, when the endometrium undergoes decidualization. This review focuses on data that support the hypothesis that paracrine signals from the rodent conceptus influence decidualization. Where possible, these findings are compared and contrasted with information currently known in other species that exhibit different implantation modes. PMID- 19551817 TI - Low rate of oral human papillomavirus (HPV) infection in women screened for cervical HPV infection in Southern Italy: A cross-sectional study of 140 immunocompetent subjects. AB - Even though the natural history of cervical and oral human papillomavirus (HPV) infection has been investigated intensely, the possibility that HPV may infect both sites in the same subject is not well documented. This study investigated the frequency of concurrent oral and cervical HPV infection in southern Italian women, in the light of some selected socio-behavioral variables. One hundred forty women (mean age: 36 years), with known cervical HPV status, were analyzed for oral HPV. Age, smoking/drinking habits, clinical and socio-behavioral history were assessed by personal interviews. Oral mucosal cells were collected by oral brushing and HPV DNA was sought by the use of nested PCR amplification followed by direct DNA sequencing and the commercial assay INNOLiPA HPV Genotyping (Innogenetics N.V., Ghent, Belgium). The data were analyzed by using the chi square test and a logistic regression (logit) model (P < 0.05 statistically significant). Oral HPV infection was detected in 2/140 (1.4%) cases, being present in 2/76 (2.6%) women with cervical HPV infection and 0/64 uninfected women (P = 0.19). A lack of type-specific concordance in the two patients with concurrent infection was observed. In the sample of population examined, HPV cervical infection does not seem to predispose to oral transmission, even in the presence of oral-genital sexual habits, thus suggesting the independence of infection at the two mucosal sites. PMID- 19551818 TI - Shared and persistent asymptomatic cutaneous human papillomavirus infections in healthy skin. AB - Cutaneous human papillomavirus (HPV) types are commonly found in normal skin, and some of them have been suspected to play a role in the development of non melanoma skin cancer. This present study is divided into three sections, the aims of this study were to examine if certain HPV-types persist over time and if HPV types are shared within families. From the first part of the study, swab samples from foreheads were collected for three longitudinal studies from one family with a newborn baby. Five specific HPV-types were isolated from the family with a newborn, with HPV-5 and FA67 being found at various time points and prevalence rates in all four members of the family. Part 2 consisted of a followed up study from two families with a 6 years interval. Six of the family members were found to have at least one of the HPV-types identified in the family 6 years earlier. Many of the HPV-types identified were shared within the families studied. Part 3 of this study involved weekly samples from four healthy females for 4 months. Among the four healthy individuals, 11%, 65%, and 56% of the weekly samples were HPV-DNA positive with one individual HPV-negative. All specimens were tested for HPV-DNA by PCR using the broad range HPV-type primer pair FAP59/64. The positive samples were HPV-type determined by cloning and sequencing. Specific cutaneous HPV-types persist over long periods of time in healthy skin in most individuals investigated and certain HPVs are shared between family members. PMID- 19551819 TI - Seroprevalence, risk factors, and hepatitis C virus genotypes in groups with high risk sexual behavior in Croatia. AB - The seroprevalence, risk factors and genotypes of hepatitis C virus (HCV) in groups with high-risk sexual behavior (persons with multiple sexual partners, men who have sex with men, commercial sex workers and their clients and persons with sexually transmitted diseases) in seven Croatian cities were analyzed. A total of 821 participants without history of injecting drug use were included in the study. Anti-HCV prevalence among risk groups varied from 2.9% to 8.5% with an overall prevalence of 4.6% (95% CI = 3.2-6.1) compared with 0.5% (95% CI = 0.0 1.5) in controls (pregnant females; OR = 9.66; 95% CI = 1.32-70.7). HCV-RNA was detected in 73.1% anti-HCV positive patients. Three of the seronegative cases (2.1%) were also found to be HCV-RNA positive ("window period"). Genotype 1 was most commonly detected (55.6%). The most prevalent subtypes were 1a (38.9%) and 3a (38.9%). Sociodemographic characteristics (age, gender, marital status and level of education) were not associated with anti-HCV seropositivity. Among sexually transmitted disease markers, a higher seroprevalence of HCV infection was found in subjects with a history of HBV infection (10.5% vs. 3.8%, P = 0.002) and gonorrhea (13.2% vs. 4.2%, P = 0.011). No other factors reflecting risk sexual behavior such as sexual orientation, number of sexual partners and number of risk behaviors were associated with HCV seroprevalence. PMID- 19551820 TI - Influenza A virus in Taiwan, 1980-2006: Phylogenetic and antigenic characteristics of the hemagglutinin gene. AB - Limited amount of information is available in Taiwan on the genetic or antigenic characteristics of influenza A virus prior to the establishment of a Taiwan surveillance network in 2000. Isolates of H1N1 and H3N2 viruses in Taiwan between 1980 and 2006 were studied, and part of the hemagglutinin gene was analyzed due to its importance in terms of viral infection and antibody neutralization. Results from a phylogenetic analysis indicate continuous evolutionary topology in H3N2 isolates, and two distinct H1N1 lineages. Many genetic relationships between vaccine strains and epidemic isolates appearing in Taiwan before other global locations were also observed and recorded in addition to a gradual increase in the number of N-linked glycosylation sites on partial HA1 proteins since 1980. The results from pairwise comparisons of HA1 nucleotide and deduced amino acid sequences indicate shared identities within groups organized according to their bootstrap and P-values of approximately 95.5-100% and 95.7-100% in H1N1 and 94.5 100% and 93.2-100% in H3N2 viruses, respectively. Comparisons of amino acid substitutions in the five antigenic regions reveal highly non-synonymous changes occurring in the Sb region of H1N1 and in the B region of H3N2. The results of an antigenic analysis using a hemagglutinin inhibition (HI) test indicate the presence of some epidemic strains 1-2 years earlier in Taiwan than in other parts of the world, as well as higher vaccine mismatch rates. This information supports the need for continuous surveillance of emerging influenza viruses in Taiwan, which will be useful for making global vaccine decisions. PMID- 19551823 TI - Adenovirus type 3 outbreak in connecticut associated with a novel variant. AB - An adenovirus outbreak occurred in New Haven, Connecticut in 2006-2007. Molecular typing revealed a twofold increase in adenovirus type 3 infections. Restriction enzyme analysis (REA) indicated that the CT outbreak was largely due to a marked increase in the novel Ad3a51 strain. This outbreak represents the first detection of Ad3a51 in the United States. While most infections were mild, children under 3 were at increased risk for severe disease and one patient with underlying disease died. PMID- 19551822 TI - Gene expression profiling of dengue infected human primary cells identifies secreted mediators in vivo. AB - We used gene expression profiling of human primary cells infected in vitro with dengue virus (DENV) as a tool to identify secreted mediators induced in response to the infection. Affymetrix GeneChip analysis of human primary monocytes, B cells and dendritic cells infected with DENV in vitro showed strong induction of monocyte chemotactic protein 2 (MCP-2/CCL8), interferon gamma-induced protein 10 (IP-10/CXCL10) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/TNFSF10). The expression of these genes was confirmed in dendritic cells infected with DENV in vitro at mRNA and protein levels. A prospectively enrolled cohort of DENV-infected Venezuelan patients was used to measure the levels of these proteins in serum during three different periods of the disease. Results showed significant increase of MCP-2, IP-10, and TRAIL levels in patients infected with DENV during the febrile period, when compared to healthy donors and patients with other febrile illnesses. MCP-2 and IP-10 levels were still elevated during the post-febrile period while TRAIL levels dropped close to normal after defervescense. Patients with primary infections had higher TRAIL levels than patients with secondary infections during the febrile period of the disease. Increased levels of IP-10, TRAIL and MCP-2 in acute DENV infections suggest a role for these mediators in the immune response to the infection. MCP-2 was identified in this work as a new unreported and important dengue-related protein and IP-10 was confirmed as a novel and strong pro-inflammatory marker in acute disease. PMID- 19551824 TI - Human T-lymphotropic virus type 1 (HTLV-1) genetic typing in Kakeroma Island, an island at the crossroads of the ryukyuans and Wajin in Japan, providing further insights into the origin of the virus in Japan. AB - Peripheral blood samples were collected from 23 human T-lymphotropic virus type-1 (HTLV-1) carriers residing in Kakeroma Island, Japan (Kagoshima Prefecture, Oshima County, Setouchi Town), one of the most highly endemic areas in Japan. The samples were subjected to amplification by PCR and sequencing of the Long Terminal Repeat in order to reconstruct a phylogenetic tree of HTLV-1 isolates. Restriction Fragment Length Polymorphism (RFLP) analysis of env region was also conducted for subgrouping of HTLV-1. Although one sample could not be amplified by PCR, and three more could not be sequenced due to the existence of conspicuous nonspecific bands or repeated sequences, the phylogenetic analysis revealed that the remaining 19 isolates obtained from Kakeroma Island belonged to either the Transcontinental or the Japanese subgroups of the Cosmopolitan subtype, one of the three major subtypes. The RFLP data corresponded closely with the typing data throughout the sequencing. The proportion of the Transcontinental subgroup among the isolates was 26.3% (5 of 19) by sequence analysis and 27.3% (6 of 22) by RFLP. Unlike in Taiwan, China and Okinawa, the Japanese subgroup was dominant in Kakeroma Island. The analysis would also suggest that the Japanese subgroup seems not to have derived from the Transcontinental subgroup, but rather that the Transcontinental subgroup came to Japan first and was followed later by the Japanese one. PMID- 19551825 TI - Development of real-time reverse transcriptase polymerase chain reaction methods for human rabies diagnosis. AB - To improve timely ante-mortem human rabies diagnosis, methods to detect viral RNA by TaqMan-based quantitative reverse transcriptase polymerase chain reactions (qRT-PCRs) have been developed. Three sets of two primers and one internal dual labeled probe for each primer set that target distinct conserved regions of the rabies virus N gene were designed and evaluated. Using a collection of 203 isolates representative of the world-wide diversity of rabies virus, all three primers/probe sets were shown to detect a wide range of rabies virus strains with very few detection failures; the RABVD1 set in particular was the most broadly reactive. These qRT-PCR assays were shown to be quantitative over a wide range of viral titer and were 100-1,000 times more sensitive than nested RT-PCR; however, both the standard and real-time PCR methods yielded concordant results when used to test a collection of archived human suspect samples. The qRT-PCR assay was employed to monitor virus load in the saliva of a rabies virus-infected patient undergoing the Milwaukee treatment protocol. However in this case it would appear that reduction of the viral load in the patient's saliva over time did not appear to correlate well with clearance of viral components from the brain. PMID- 19551826 TI - Clearance of serum HBsAg and anti-HBs seroconversion following antiviral therapy for chronic hepatitis B. AB - In this study, we have analyzed the evolution of serum HBsAg levels in 16 patients with chronic hepatitis B who showed an HBsAg seroconversion following antiviral therapy. The data showed that the clearance of serum HBsAg is slower than that of serum HBV DNA, which may reflect a slow kinetics of clearance of infected hepatocytes. Interestingly, HBsAg was detectable for a longer time using the Architect assay than with the Bio-Rad assay. As viremia suppression is achieved in most patients under therapy with the new generation of nucleoside analogs, these data suggest that the quantitative monitoring of serum HBsAg may represent a novel tool for the assessment of antiviral therapy efficacy. PMID- 19551827 TI - Genomic mutations of viral protein 1 and BK virus nephropathy in kidney transplant recipients. AB - Genomic variability in the viral protein 1 region of BK polyomavirus (BKV) may change the ability of the virus to replicate. The significance of such changes was studied in clinical samples taken from kidney transplant patients with and without BKV nephropathy. A 94 base-pair fragment of viral protein 1 was amplified from 68 urine, 28 blood, and 12 renal biopsy samples from eight patients with BKV nephropathy, and from 100 urine samples, 17 blood and three renal biopsy samples from 41 of 218 controls. The DNA was sequenced and the amino acid changes were predicted by the Expert Protein Analysis System program (ExPASy, Swiss Institute of Bioinformatics, Geneva, Switzerland). Single base-pair mutations were detected more frequently in the samples from the BKV nephropathy patients than in the controls, and this was the only statistically significant finding of the study (P < 0.05), thus suggesting a greater genetic instability in BKV nephropathy associated strains. The amino acid changes were distributed at random in both BKV nephropathy patients and controls. However, one aspartic acid-to-asparagine substitution at residue 75 was detected in all samples of the one patient with BKV-associated nephropathy, who developed disease progression confirmed by histology, and not in any of the other patient or control samples. Whether this specific amino acid change plays a role in disease deserves further study. PMID- 19551828 TI - Differences in viral kinetics between genotypes 1 and 2 of hepatitis C virus after single administration of standard interferon-alpha. AB - Hepatitis C virus (HCV) kinetics were determined after a single administration of standard interferon (IFN) according to the HCV genotype that affects the response to antiviral therapy with IFN/peginterferon. A total of 208 patients were investigated. All patients received a single administration of 6 megaunits of standard IFN-alpha. HCV RNA concentration was measured before, and at 24, 48, 72, and 120 hr after administration. Changes in HCV RNA concentration were compared between genotypes. The patient group consisted of 132 patients with genotype 1B, 58 with genotype 2A, and 18 with genotype 2B. In the comparison between genotypes 1 and 2, the reduction in HCV RNA concentration after a single IFN administration was less marked in patients with genotype 1B at both 24 and 48 hr after administration (P < 0.0001). In contrast, an increase in HCV RNA concentration during 24-48 or 24-72 hr after a single administration was comparable between genotypes 1 and 2. In the comparison between genotypes 2A and 2B, the reduction in HCV RNA concentration after a single administration was more marked in patients with genotype 2A, despite the similar rate of sustained virologic response to peginterferon and ribavirin combination therapy. The results of the study indicate that the rapid decrease in HCV RNA concentration observed during IFN or peginterferon therapy in patients with genotype 2 appeared to be due to the difference in sensitivity to IFN. Within genotype 2, HCV genotype 2A was more sensitive to IFN than genotype 2B. PMID- 19551829 TI - Use of stored dried blood spots for retrospective diagnosis of congenital CMV. AB - The diagnosis of congenital cytomegalovirus infection cannot be made with certainty in children presenting after the perinatal period, unless stored early samples are available for diagnostic testing. This has led to uncertainty in confirming the overall contribution of CMV to hearing loss and neurodevelopmental impairment. The use of dried blood spots (DBSs) to retrospectively diagnose infection in children with compatible symptoms may be helpful diagnostically although there are ongoing uncertainties regarding the stability of viral DNA in cards, the risk of contamination between cards, and sensitivity and specificity in a clinical setting. This report aims to address these areas and evaluate the use of DBS testing in our hands in the United Kingdom to date. Results from testing artificially prepared cards and cards from three populations of children suggest a high specificity for congenital CMV infection and a good sensitivity for cases where sensorineural hearing loss is caused by congenital CMV. PMID- 19551830 TI - Lower cellular immune responses to influenza A (H3N2) in the elderly. AB - Influenza epidemic is an important cause of severe illness in the elderly. Age dependent morbidity of influenza in the elderly is associated with weakened immunity. The baseline age-related memory T cell activity in Chinese persons who are exposed to influenza virus through natural infection, are associated with the protective response to the virus after vaccination, thus providing important pre vaccination information. A cohort from the general population was established at the end of an influenza season in an area where influenza occurs regularly, and followed for 24 weeks. The subjects had no vaccination history for 5 years. Memory T cell responses were evaluated using a set of peptides spanning the influenza A (H3N2) entire proteome in a gamma interferon (IFN-gamma)-enzyme linked immunospot (ELISPOT) assay, prior to the next influenza season. Changes of hemagglutination inhibition (HI) antibody titers were also evaluated. IFN gamma(+) T cell responses against influenza peptides were significantly lower in subjects of 60 years and older. Although the age-related decline of cellular immune response was clear, no significant association of antibody titers with age was found. The pre-vaccination baseline of memory IFN-gamma(+) T cell immunity state in elderly Chinese was significantly lower than in people younger than 60 years. Measurement of the ex vivo cellular immune responses to influenza should be incorporated into the evaluation of protective immunity in elderly persons. PMID- 19551831 TI - Correlation of rhinovirus load in the respiratory tract and clinical symptoms in hospitalized immunocompetent and immunocompromised patients. AB - While human rhinoviruses (HRVs) are well accepted as a major cause of common cold syndromes (rhinitis), their role in the etiology of lower respiratory tract infections is still controversial, and their detection in asymptomatic patients is relatively common. The HRV pathogenic role in four groups of hospitalized patients (pediatric immunocompetent and immunocompromised patients, and adult immunocompetent and immunocompromised patients) was investigated by quantifying HRV load in nasopharyngeal aspirates or bronchoalveolar lavage samples by real time reverse transcription PCR (RT-PCR). Real-time RT-PCR was performed in duplicate on all respiratory samples resulting positive by qualitative RT-PCR. In addition, molecular typing allowed detection of all known HRV species (A, B, and C). In immunocompetent pediatric patients HRVs were mostly associated with lower respiratory tract infections (in the absence of other viral agents) and wheezing, when viral load was > or =10(6) RNA copies/ml. In young immunocompromised patients (stem cell transplantation recipients), an inverse correlation between HRV persistence over time and time at which the infection occurred after transplantation was observed, whereas in adult immunocompromised patients (lung transplant recipients) HRVs could be detected at a medium-low level (<10(5) RNA copies/ml) in bronchoalveolar lavage samples taken routinely from asymptomatic patients. In conclusion, when detected at high viral load, HRVs may cause severe upper and lower respiratory tract infections, whereas when detected at a medium low viral load, an event more frequent in immunocompromised subjects, they may represent only bystander viruses. PMID- 19551834 TI - Molecular detection of hepatitis B, hepatitis C, and torque teno viruses in drug users in Saudi Arabia. AB - Injecting drug users are at increased risk of infection with hepatitis viruses and blood-borne pathogens. The aim of this study was to examine HBV, HCV, HDV, and TTV infections in Saudi drug users (N = 344). Extraction of nucleic acid from serum, reverse-transcription, amplification of viral nucleic acids, and HBV and HCV genotyping were done using established techniques. Of the analyzed samples, 41 (12%) contained detectable HBV DNA, 131 (38%) contained detectable HCV RNA, and 174 (51%) had detectable TTV DNA. The predominant HBV genotype was found to be genotype D and the predominant HCV genotype was found to be genotype 1b. All the samples were negative for HDV. Twelve samples (3.5%) were found to contain mixed HBV and HCV genomes, 24 samples (7%) were found to contain mixed HBV and TTV genomes, 82 samples (24%) were found to contain mixed HCV and TTV genomes, and 9 samples (2.6%) were found to contain mixed HBV, HCV, and TTV genomes. Identification of various infections in drug users will help the control of these infections in this group as well as in the community. PMID- 19551833 TI - Monitoring of herpes simplex virus DNA types 1 and 2 viral load in cerebrospinal fluid by real-time PCR in patients with herpes simplex encephalitis. AB - A quantitative polymerase chain reaction (PCR) assay was evaluated retrospectively on 92 cerebrospinal fluid (CSF) samples from 29 patients with herpes simplex virus (HSV) encephalitis with the aim to study if the concentration of HSV genomes can be used as a prognostic marker and for monitoring of antiviral therapy. The results were compared to those obtained previously by nested PCR, and the numbers of HSV genomes/ml were evaluated in correlation to patient outcome and treatment. The aims were to compare the sensitivity of a conventional nested PCR to a quantitative PCR, to investigate the range of HSV genome concentration in initial samples and to evaluate possible relationships between the HSV DNA concentrations in CSF, neopterin levels, and outcome of disease. The 29 initial samples contained between 2 x 10(2) and 42 x 10(6) HSV genomes/ml. There was no apparent correlation between the amount of HSV DNA in the initial samples and income status, initial neopterin levels, or prognosis. The number of HSV genomes/ml declined after treatment in all patients, but HSV DNA was still detectable after day 20 in 3 out of 16 patients. A long duration of genome detectability was found to correlate with poor outcome. There was no difference in sensitivity between the nested PCR and the quantitative PCR. While the quantitative PCR is more rational than a nested PCR, the quantitation of HSV genomes does not seem very useful as a prognostic marker in HSV encephalitis. PMID- 19551832 TI - Seroprevalence of Kaposi's sarcoma-associated herpesvirus and risk factors in Xinjiang, China. AB - Xinjiang, China is an endemic area for Kaposi's sarcoma (KS) but the seroprevalence of Kaposi's sarcoma-associated herpesvirus (KSHV) and risk factors remain undefined. In this study, antibodies to one KSHV latent protein (ORF73) and two KSHV lytic proteins (ORF65 and ORF-K8.1) were examined in 2,228 subjects from the general population and 37 subjects infected with HIV-1 in Xinjiang, and 560 subjects from the general population in Hubei, a low KS incidence region. The serostatus of a serum sample was defined based on positive results in any one of the three serologic assays. The seroprevalence of KSHV in the general population was higher in Xinjiang than in Hubei (19.2% vs. 9.5%; odds ratios [OR], 2.28; 95% confidence interval [CI], 1.68-3.08; P < 0.001). Among the ethnic groups in Xinjiang, 68 (15.8%) Han, 182 (20.7%) Uygur, 140 (19.9%) Hazakh, 9 (33.3%) Xibo, and 29 (16.8%) Hui were KSHV-seropositive, respectively. Compared to the Han, the latter groups had an increase in the risk of KSHV of 62.2%, 63.8%, 180.1%, and 30.2% (P = 0.003, 0.004, 0.018, and 0.286, respectively). Subjects aged <20, 20 50, and >50 had a seroprevalence of KSHV of 11.8%, 17.9%, and 24.6%, respectively. Compared to subjects aged <20, the latter groups had an increase in the risk of KSHV of 63.3% and 144.5% (P = 0.009 and <0.001, respectively). Subjects infected with HIV-1 in Xinjiang had a seroprevalence of KSHV of 43.2%, and a 220% increase in the risk of KSHV compared to the general population (P < 0.001). Similar results were obtained when the seroprevalence of KSHV was analyzed with any single or two of the three serologic assays alone. Genotyping identified three unique sequences clustered in the A clade. This study indicates that Xinjiang has a high seroprevalence of KSHV. Geographic location, ethnicity, age and HIV-1 infection are risk factors. Serologic and genotyping results suggest the introduction of KSHV into Xinjiang by specific ethnic groups. PMID- 19551835 TI - Cytomegalovirus and Epstein-Barr virus coinfection in three toddlers with prolonged illnesses. AB - Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) usually cause primary and latent infections during childhood; thus, coinfection with these viruses occurs occasionally in children. However, its clinical impact has not been established, and may be underestimated. Three cases of coinfection involving these two viruses in toddlers are described: a 14-month-old male with infectious mononucleosis, an 18-month-old female with hemophagocytic lymphohistiocytosis, and a 13-month-old female with acute hepatitis. All three patients had prolonged illnesses. Serial serological testing and quantitation of viral DNA for CMV and EBV using peripheral blood from the patients suggested primary infections with both viruses. In all three cases, the viral load of EBV and CMV in the early stage of disease exceeded 6.4 x 10(3) and 8.8 x 10(2) copies/ml of whole blood, respectively, suggesting that the viruses were associated with the clinical condition. Recognizing that coinfection with these viruses may modulate the clinical course of disease is important. PMID- 19551836 TI - IgA antibodies against the Epstein-Barr nuclear antigen1 as a valuable biomarker for the diagnosis of nasopharyngeal carcinoma in Tunisian patients. AB - Serological tests for Epstein-Barr virus (EBV) have been used for many years as diagnostic predictors of nasopharyngeal carcinoma. It has been shown previously that the conventional immunofluorescence assay has a limited diagnostic value, especially in young patients from North African area. In the search for more reliable immunoglobulin (Ig) G or IgA antibody markers for the diagnosis of nasopharyngeal carcinoma, immunoblot analysis was performed using a full spectrum of EBV proteins. Sera were collected from 108 patients with nasopharyngeal carcinoma and three control groups composed of 18 patients with lymphoma, 18 other patients with autoimmune diseases and 55 healthy EBV carriers. It was observed that the IgA Epstein-Barr nuclear antigen 1 (EBNA1), IgA early antigen (EA)-p138 and IgG EA-p138 antibodies represent the most specific anti-EBV responses in either young or older patients with nasopharyngeal carcinoma which yield higher positive rates compared to the three control groups. Since the IgA EBNA1 response showed the highest sensitivity value for the detection of nasopharyngeal carcinoma, a novel enzyme-linked immunosorbent assay (ELISA) was established using a GST-EBNA1 protein expressed in bacteria, containing the P threonine EBNA1 subtype cloned from DNA EBV sequence of C15 xenograft cells. Detection rates were 85.7% and 94.9% in young and older patients with nasopharyngeal carcinoma respectively, while only 3.6%, 11.1%, and 16.6% in healthy EBV carriers, patients with lymphoma and patients with autoimmune diseases, respectively. Thus, IgA EBNA1 ELISA may be useful for early diagnosis and mass screening of nasopharyngeal carcinoma in Tunisia even in young patients. PMID- 19551837 TI - Genetic characterization of Chinese measles vaccines by analysis of complete genomic sequences. AB - The complete genomic sequences of two Chinese measles vaccine viruses, Shanghai 191 (S-191) and Changchun-47 (C-47), were determined and compared to the sequences of other measles vaccine strains as well as the prototype measles strain, Edmonston wild-type (Edwt). Compared to Edwt, S-191 and C-47 had 49 and 43 nucleotide changes, respectively. These differences were found at 52 nucleotide positions that were not found in other vaccine strains. Phylogenetic analysis of the all of the available genomic sequences for measles vaccines showed that S-191 and C-47 were most closely related to the Leningrad-4 strain. S 191 and C-47 shared conserved vaccine virus-specific amino acid changes in the phosphoprotein (P), V, C, matrix (M), and hemagglutinin (H) that could represent important targets for future studies aimed at understanding the molecular basis of attenuation. In addition, S-191 and C-47 had several unique amino acid changes including 13 positions that differed from Edwt. This is the first comparison of the complete genomic sequences of Chinese measles vaccines to the sequences of other vaccine strains. PMID- 19551838 TI - A novel genotype of hepatitis E virus prevalent among farmed rabbits in China. AB - In total, 335 serum samples were collected from rabbits from two farms in Gansu province, China, and tested for anti-hepatitis E virus (HEV) antibody using EIA and for HEV RNA using nested RT- PCR with ORF2 primers. The overall prevalence of anti-HEV antibody and HEV RNA was 57.0% (191/335) and 7.5% (25/335), respectively. The positivity rate of HEV RNA in the anti-HEV antibody negative group (7.6% (11/144)) did not differ significantly from that in the positive group (7.3% (14/191)). The concordance between HEV RNA and anti-HEV antibody was 43.3% with no significant correlation (P < 0.05). All 25 amplicons from the ORF2 region were cloned and sequenced. On the basis of nucleotide sequence comparison, they had 84-99% identity to each other and 73-77%, 70-76%, 75-82%, 71-77%, and 53 65% with the corresponding regions of genotypes 1, 2, 3, 4, and avian HEV, respectively. Samples that were positive with the ORF2 primers were amplified using ORF1 region primers; 17 were positive and shared 71-78%, 73-76%, 74-82%, 72 78%, and 39-58% identity with the corresponding regions of genotypes 1, 2, 3, 4, and avian HEV, respectively, at the nucleotide level. Two representative full length sequences were determined. These two sequences shared 85% identity with each other and had 74%, 73%, 78-79%, 74-75%, and 46-47% identity to full-length genotypes 1, 2, 3, 4, and avian HEV, respectively. Thus, the sequences isolated from the rabbits represent a novel genotype of HEV. This study provides novel information about HEV genotypes infecting rabbits as well as evidence of a new mammalian genotype of HEV. PMID- 19551839 TI - Molecular analysis of transmission of hepatitis C virus in a nurse who acquired acute hepatitis C after caring for a viremic patient with epistaxis. AB - A 23-year-old nurse (HC-IP) developed acute hepatitis C. Intrafamilial transmission of hepatitis C virus (HCV) was suspected initially because her parents were carriers of HCV of the same genotype (1b) as that of Patient HC-IP. However, the HCV isolate from Patient HC-IP and those from her parents shared identities of only 92.4-92.7% in the 1,087-nucleotide (nt) sequence within the NS5B region. It was then suspected that she contracted HCV infection during medical practice. Sixteen patients with antibodies to HCV (anti-HCV) were hospitalized 1-3 months before she became positive for anti-HCV. Upon analysis of stored serum samples, 14 of the 16 patients were found to be positive for HCV RNA, and 9 of the 14 viremic patients had genotype 1b HCV. Although the shared identities between the HCV isolate from Patient HC-IP and those from eight of the nine patients were merely 90.6-93.9% within the 1,087-nt NS5B sequence, the HCV isolate from the remaining one patient (HC-P12) was 99.7% identical to that from Patient HC-IP. Upon analysis of the E1 and E2 junctional region including hypervariable region 1 (283 nt), there was a close relationship (99.3-100%) between clones obtained from Patients HC-IP and HC-P12. Although the nurse HC-IP had a finger injury, she took care of Patient HC-P12, a 70-year-old man with HCV related cirrhosis and recurrent epistaxis, occasionally without wearing protective gloves. This study indicates the occurrence of HCV transmission by exposure of nonintact skin to blood in health care settings. PMID- 19551840 TI - Temperature-dependent release of volatile organic compounds of eucalypts by direct analysis in real time (DART) mass spectrometry. AB - A method is described for the rapid identification of biogenic, volatile organic compounds (VOCs) emitted by plants, including the analysis of the temperature dependence of those emissions. Direct analysis in real time (DART) enabled ionization of VOCs from stem and leaf of several eucalyptus species including E. cinerea, E. citriodora, E. nicholii and E. sideroxylon. Plant tissues were placed directly in the gap between the DART ionization source skimmer and the capillary inlet of the time-of-flight (TOF) mass spectrometer. Temperature-dependent emission of VOCs was achieved by adjusting the temperature of the helium gas into the DART ionization source at 50, 100, 200 and 300 degrees C, which enabled direct evaporation of compounds, up to the onset of pyrolysis of plant fibres (i.e. cellulose and lignin). Accurate mass measurements facilitated by TOF mass spectrometry provided elemental compositions for the VOCs. A wide range of compounds was detected from simple organic compounds (i.e. methanol and acetone) to a series of monoterpenes (i.e. pinene, camphene, cymene, eucalyptol) common to many plant species, as well as several less abundant sesquiterpenes and flavonoids (i.e. naringenin, spathulenol, eucalyptin) with antioxidant and antimicrobial properties. The leaf and stem tissues for all four eucalypt species showed similar compounds. The relative abundances of methanol and ethanol were greater in stem wood than in leaf tissue suggesting that DART could be used to investigate the tissue-specific transport and emissions of VOCs. PMID- 19551841 TI - Timp-2 binding with cellular MT1-MMP stimulates invasion-promoting MEK/ERK signaling in cancer cells. AB - Both invasion-promoting MT1-MMP and its physiological inhibitor TIMP-2 play a significant role in tumorigenesis and are identified in the most aggressive cancers. Despite its antiproteolytic effects in vitro, clinical data suggest that TIMP-2 expression is positively associated with tumor recurrence, thus emphasizing the wide-ranging role of TIMP-2 in malignancies. To shed light on this role of TIMP-2, we report that low concentrations of TIMP-2, by interacting with MT1-MMP (a specific membrane receptor of TIMP-2), induce the MEK/ERK signaling cascade in fibrosarcoma HT1080 cells which express MT1-MMP naturally. TIMP-2 binding with cell surface-associated MT1-MMP stimulates phosphorylation of MEK1/2, which is upstream of ERK1/2, and the ERK1/2 substrate p90RSK. Consistent with volumes of literature, we confirmed that the activation of ERK stimulated cell migration. Both the transcriptional silencing of MT1-MMP and the inhibition of MEK1/2 reversed the signaling effects of TIMP-2/MT1-MMP while the active site targeting MMP inhibitor GM6001 did not. Our data suggest that both the interactions of TIMP-2 with MT1-MMP, which activate the pro-migratory ERK signaling cascade,and the conventional inhibition of MT1-MMP's catalytic activity by TIMP-2, play a role in the invasion-promoting function of MT1-MMP. The TIMP-2 induced stimulation of ERK signaling in cancer cells explains the direct, as opposed to the inverse, association of TIMP-2 expression with poor prognosis in cancer. PMID- 19551843 TI - An engineered tryptophan zipper-type peptide as a molecular recognition scaffold. AB - In an effort to develop a structured peptide scaffold that lacks a disulfide bond and is thus suitable for molecular recognition applications in the reducing environment of the cytosol, we investigated engineered versions of the trpzip class of beta-hairpin peptides. We have previously shown that even most highly folded members of the trpzip class (i.e. the 16mer peptide HP5W4) are substantially destabilized by the introduction of mutations in the turn region and therefore not an ideal peptide scaffold. To address this issue, we used a FRET-based live cell screening system to identify extended trpzip-type peptides with additional stabilizing interactions. One of the most promising of these extended trpzip-type variants is the 24mer xxtz1-peptide with the sequence KAWTHDWTWNPATGKWTWLWRKNK. A phage display library of this peptide with randomization of six residues with side chains directed towards one face of the hairpin was constructed and panned against immobilized streptavidin. We have also explored the use of xxtz1-peptide for the presentation of an unstructured peptide 'loop' inserted into the turn region. Although NMR analysis provided no direct evidence for structure in the xxtz1-peptide with the loop insertion, we did attempt to use this construct as a scaffold for phage display of randomized peptide libraries. Panning of the resulting libraries against streptavidin resulted in the identification of peptide sequences with submicromolar affinities. Interestingly, substitution of key residues in the hairpin-derived portion of the peptide resulted in a 400-fold decrease in K(d), suggesting that the hairpin-derived portion plays an important role in preorganization of the loop region for molecular recognition. PMID- 19551842 TI - Dietary lycopene and tomato extract supplementations inhibit nonalcoholic steatohepatitis-promoted hepatocarcinogenesis in rats. AB - Epidemiological and experimental studies provide supportive evidence that lycopene (LY), a major carotenoid from tomatoes and tomato products, may act as a chemopreventive agent against certain types of cancers. We recently showed that high-fat diet (HFD)-induced nonalcoholic steatohepatitis (NASH) promoted diethylnitrosamine (DEN)-initiated hepatocarcinogenesis in a rat model. Using this model, we investigated the efficacy of an equivalent dosage of dietary LY from either a pure compound or a tomato extract (TE) against NASH-promoted hepatocarcinogenesis. Six groups of rats were injected with DEN and then fed either Lieber-DeCarli control diet or HFD with or without LY or TE for 6 weeks. Results showed that both LY and TE supplementations significantly decreased the number of altered hepatic foci expressing the placental form of glutathione S transferase in the livers of HFD-fed rats. This was associated with significantly lower proliferating cell nuclear antigen positive hepatocytes and cyclinD1 protein, as well as decreased activation of extracellular signal-regulated kinase and nuclear NF-kappaB. Although both LY and TE supplementations reduced HFD induced lipid peroxidation in the livers, we observed significantly decreased cytochrome P450 2E1, inflammatory foci and mRNA expression of proinflammatory cytokines (TNF-alpha, IL-1beta and IL-12) in the HFD+TE fed group but increased nuclear NF-E2-related factor-2 and heme oxygenase-1 proteins in the HFD+LY fed group, relative to HFD feeding alone. These data indicate that LY and TE can inhibit NASH-promoted hepatocarcinogenesis mainly as a result of reduced oxidative stress, which could be fulfilled through different mechanisms. PMID- 19551844 TI - Myeloid derived suppressor cells inhibit natural killer cells in patients with hepatocellular carcinoma via the NKp30 receptor. AB - Several immune suppressive mechanisms that evade the host immune response have been described in patients with hepatocellular carcinoma (HCC); one of these mechanisms is expansion of myeloid-derived suppressor cells (MDSCs). MDSCs have been shown to inhibit T cell responses in tumor-bearing mice, but little is known about these cells in humans. Here, we have analyzed and characterized the effect of MDSCs on the innate immune system, in particular, their interaction with natural killer (NK) cells in patients with HCC. MDSCs from patients with HCC inhibited autologous NK cell cytotoxicity and cytokine secretion when cultured together in vitro. This suppression was dependent on cell contact, but did not rely on the arginase activity of MDSCs, which is a hallmark function of these cells. However, MDSC-mediated inhibition of NK cell function was dependent mainly on the NKp30 on NK cells. CONCLUSION: Our study suggests a new role for MDSCs in patients with HCC in disarming the innate immune system and further contributing to the immune suppressor network in these patients. These findings have important implications when designing immunotherapy protocols. PMID- 19551845 TI - A binary matrix for improved detection of phosphopeptides in matrix-assisted laser desorption/ionization mass spectrometry. AB - Application of matrix-assisted laser-desorption/ionization mass spectrometry (MALDI MS) to analysis and characterization of phosphopeptides in peptide mixtures may have a limitation, because of the lower ionizing efficiency of phosphopeptides than nonphosphorylated peptides in MALDI MS. In this work, a binary matrix that consists of two conventional matrices of 3-hydroxypicolinic acid (3-HPA) and alpha-cyano-4-hydroxycinnamic acid (CCA) was tested for phosphopeptide analysis. 3-HPA and CCA were found to be hot matrices, and 3-HPA not as good as CCA and 2,5-dihydroxybenzoic acid (DHB) for peptide analysis. However, the presence of 3-HPA in the CCA solution with a volume ratio of 1:1 could significantly enhance ion signals for phosphopeptides in both positive-ion and negative-ion detection modes compared with the use of pure CCA or DHB, the most common phosphopeptide matrices. Higher signal intensities of phosphopeptides could be obtained with lower laser power using the binary matrix. Neutral loss of the phosphate group (-80 Da) and phosphoric acid (-98 Da) from the phosphorylated residue-containing peptide ions with the binary matrix was decreased compared with CCA alone. In addition, since the crystal shape prepared with the binary matrix was more homogeneous than that prepared with DHB, searching for 'sweet' spots can be avoided. The sensitivity to detect singly or doubly phosphorylated peptides in peptide mixtures was higher than that obtained with pure CCA and as good as that obtained using DHB. We also used the binary matrix to detect the in solution tryptic digest of the crude casein extracted from commercially available low fat milk sample, and found six phosphopeptides to match the digestion products of casein, based on mass-to-charge values and LIFT TOF-TOF spectra. PMID- 19551847 TI - Immunohistochemical detection of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 expression in breast carcinomas: comparison on cell block, needle-core, and tissue block preparations. AB - BACKGROUND: Fine-needle aspiration (FNA) is a rapid and accurate procedure for the detection of breast carcinomas. The evaluation of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) expression by immunohistochemistry (IHC) is performed routinely on formalin fixed, paraffin-embedded needle-core (NC) or excision tissue block (TB) preparations, according to the American Society of Clinical Oncology/College of American Pathologist guidelines. In this retrospective study, the authors compared expression levels of ER, PR, and HER2 in ethanol-fixed BC FNA cell block (CB) samples with expression levels in formalin-fixed NC and TB samples. METHODS: Forty-one breast carcinoma CB samples with concurrent or subsequent NC and TB samples were identified. Patients who had received neoadjuvant or adjuvant chemotherapy were excluded. CB samples initially were fixed in 50% ethanol (4-12 hours), and this was followed by formalin fixation (minimum, 6 hours). NC samples were placed promptly in formalin for a minimum of 6 hours. Within 4 to 8 hours, TB samples were fixed in formalin for 6 to 48 hours. Fluorescence in situ hybridization (FISH) results were also compared. RESULTS: IHC for ER on alcohol fixed CB samples had good correlation with NC and TB samples. PR results on TB samples had excellent agreement with NC samples. A higher discordance rate wais observed when PR results were compared between CB samples and NC samples. HER2 detection on ethanol-fixed CB samples resulted in a higher rate of positive and equivocal staining than NC or TB samples. HER2 IHC on TB samples demonstrated better correlation with FISH results than CB or NC samples. CONCLUSIONS: Alcohol fixation did not affect ER results in breast carcinoma, but it may alter tumor cell PR antigenicity. The authors concluded that CB samples could be used to triage patients for tamoxifen therapy, but they are not reliable for the assessment of HER2 status; therefore, CB results should be correlated with results from NC or TB samples. PMID- 19551846 TI - Semi-quantitative and structural metabolic phenotyping by direct infusion ion trap mass spectrometry and its application in genetical metabolomics. AB - The identification of quantitative trait loci (QTL) for plant metabolites requires the quantitation of these metabolites across a large range of progeny. We developed a rapid metabolic profiling method using both untargeted and targeted direct infusion tandem mass spectrometry (DIMSMS) with a linear ion trap mass spectrometer yielding sufficient precision and accuracy for the quantification of a large number of metabolites in a high-throughput environment. The untargeted DIMSMS method uses top-down data-dependent fragmentation yielding MS(2) and MS(3) spectra. We have developed software tools to assess the structural homogeneity of the MS(2) and MS(3) spectra hence their utility for phenotyping and genetical metabolomics. In addition we used a targeted DIMS(MS) method for rapid quantitation of specific compounds. This method was compared with targeted LC/MS/MS methods for these compounds. The DIMSMS methods showed sufficient precision and accuracy for QTL discovery. We phenotyped 200 individual Lolium perenne genotypes from a mapping population harvested in two consecutive years. Computational and statistical analyses identified 246 nominal m/z bins with sufficient precision and homogeneity for QTL discovery. Comparison of the data for specific metabolites obtained by DIMSMS with the results from targeted LC/MS/MS analysis showed that quantitation by this metabolic profiling method is reasonably accurate. Of the top 100 MS(1) bins, 22 ions gave one or more reproducible QTL across the 2 years. PMID- 19551848 TI - Interinstitutional consultation in fine-needle aspiration cytopathology: a study of 742 cases. AB - BACKGROUND: The importance of interinstitutional consultation (IC) has been documented across a variety of surgical pathology organ systems. However, to the authors' knowledge, few studies exist regarding this practice within cytopathology and specifically within fine-needle aspiration cytology (FNAC). METHODS: All FNAC cases between September 2002 and January 2007 were reviewed. Original diagnoses and second opinion diagnoses (SODs) were categorized as either no diagnostic disagreement, or minor diagnostic disagreement, or major diagnostic disagreement, and the latter was defined as either a 2-step deviation on a scale of "unsatisfactory, benign, atypical, suspicious, and malignant" or a change in treatment and/or prognosis. Outcome was determined by a review of the electronic medical record. RESULTS: Among 742 FNAC cases from outside laboratories, there were minor disagreements in 132 cases (17.8%) and major disagreements in 69 cases (9.3%) compared with the SODs from the authors' laboratory. Follow-up was available for 60 of 69 major discrepancies. The SOD was supported on follow-up in 65% of major discrepancies, and the initial diagnosis was supported better in 33% of major discrepancies. In 55% of cases in which the original institution diagnosis was supported better, either no case slides were received for examination or the slides contained material that was considered nondiagnostic by the authors. An SOD prompted a change in clinical management in 32 of 742 patients (4.3%). Aspirates that were most prone to change in management or therapy were from the thyroid (13 cases), neck (soft tissue and lymph nodes; 9 cases), salivary gland (2 cases), and liver (2 cases). Of 60 major diagnostic disagreements, board-certified cytopathologists rendered an SOD in 44 cases, and 75% of the diagnoses were supported better by follow-up, whereas pathologists who were not board certified in cytopathology had only 38% of SODs supported. CONCLUSIONS: Of 742 FNAC cases, 9.3% had major diagnostic disagreements; and, in 4.3%, patient management and therapy were altered. These results were similar to studies in surgical pathology emphasizing the importance of IC in FNAC. The authors concluded that FNAC IC benefits patient care. PMID- 19551850 TI - Prediction of deep neck abscesses by contrast-enhanced computerized tomography in 76 clinically suspect consecutive patients. AB - OBJECTIVES/HYPOTHESIS: Contrast-enhanced computerized tomography (CECT) has become the imaging method of choice in patients with clinical suspicion of a deep neck abscess. The purpose of this retrospective study was to assess the predictive value of the diagnosis of deep neck abscess using CECT. STUDY DESIGN: Retrospective chart review study over a 5-year period from 2002 to 2007 by a blinded observer. METHODS: CECT examinations of patients with clinical suspicion of a deep neck abscess were reviewed for the presence of fluid collections, rim enhancement, and abnormal air collections, which we considered as indicative for an abscess. Additionally, the location and extension of the abnormalities were documented. RESULTS: In this series of 76 patients, 61 adults and 15 children, fluid collections with complete or partial rim enhancement were seen in 65 patients. In two patients large abnormal air collections were seen with nonenhancing fluid accumulation in all deep spaces. Final diagnosis was confirmed by surgery in 57 patients. In 36 patients, two or more adjacent compartments were involved. Nine patients with an abscess were treated successfully by intravenous antibiotics only. The positive predictive value (PPV) for the presence of an abscess was 82% (53/65) if all radiological signs were included. The PPV was 67% (8/12) for fluid collections without rim enhancement. Air within or adjacent to a fluid collection or excessive free air between the fascias indicated an abscess in all cases. Negative predictive value (prediction of no abscess) was 100%. CONCLUSIONS: The predictive value of the diagnosis deep neck abscess by CECT is high, with a PPV of 82%. The presence of air indicates abscess in all cases. PMID- 19551849 TI - Postischemic treatment of neonatal cerebral ischemia should target autophagy. AB - OBJECTIVE: To evaluate the contributions of autophagic, necrotic, and apoptotic cell death mechanisms after neonatal cerebral ischemia and hence define the most appropriate neuroprotective approach for postischemic therapy. METHODS: Rats were exposed to transient focal cerebral ischemia on postnatal day 12. Some rats were treated by postischemic administration of pan-caspase or autophagy inhibitors. The ischemic brain tissue was studied histologically, biochemically, and ultrastructurally for autophagic, apoptotic, and necrotic markers. RESULTS: Lysosomal and autophagic activities were increased in neurons in the ischemic area from 6 to 24 hours postinjury, as shown by immunohistochemistry against lysosomal-associated membrane protein 1 and cathepsin D, by acid phosphatase histochemistry, by increased expression of autophagosome-specific LC3-II and by punctate LC3 staining. Electron microscopy confirmed the presence of large autolysosomes and putative autophagosomes in neurons. The increases in lysosomal activity and autophagosome formation together demonstrate increased autophagy, which occurred mainly in the border of the lesion, suggesting its involvement in delayed cell death. We also provide evidence for necrosis near the center of the lesion and apoptotic-like cell death in its border, but in nonautophagic cells. Postischemic intracerebroventricular injections of autophagy inhibitor 3 methyladenine strongly reduced the lesion volume (by 46%) even when given >4 hours after the beginning of the ischemia, whereas pan-caspase inhibitors, carbobenzoxy-valyl-alanyl-aspartyl(OMe)-fluoromethylketone and quinoline-val asp(OMe)-Ch2-O-phenoxy, provided no protection. INTERPRETATION: The prominence of autophagic neuronal death in the ischemic penumbra and the neuroprotective efficacy of postischemic autophagy inhibition indicate that autophagy should be a primary target in the treatment of neonatal cerebral ischemia. PMID- 19551851 TI - Sialendoscopy-based diagnosis and classification of parotid duct stenoses. AB - OBJECTIVES/HYPOTHESIS: To assess stenoses of Stensen's duct considering findings provided by sialendoscopy-based direct visualization. A practical classification scheme based on different parameters is presented. METHODS: Ultrasonography and sialendoscopy were used to diagnose and analyze symptomatic stenoses of the parotid gland excretory duct in 93 patients. Sialendoscopy was performed with semirigid endoscopes that enabled direct evaluation and classification of those stenoses in terms of tissue quality, luminal narrowing, extend, number, location, and in consideration of possible etiologic diseases or conditions. RESULTS: A total of 111 stenoses were diagnosed in 93 patients. Three types could be distinguished. Stenoses were inflammation-dominated with various narrowing of the lumen (type I) in 16.1% of patients, fibrous and web-associated, predominant incomplete (luminal narrowing <50%, type II) in 18.3%, and fibrous, predominant high-grade (luminal narrowing >50%, type III) in 66.6%. A length of more than 1 cm or diffuse involvement of the duct system was observed in 12.9% of cases. Together 70.1% of all stenoses were found in middle and distal regions of the duct. Multiple stenoses were found in 12.9% of patients, bilateral in 6.5%. Of all patients, 45.2% had diseases possibly implicated in the etiogenesis. Type III stenoses were associated with these comorbidities in up to 100%. CONCLUSIONS: Minimally invasive techniques play a central role in the diagnosis and treatment of Stensen's duct stenoses. Sialendoscopy is the diagnostic method of first choice. It enables an exact and direct classification of stenoses and provides additional information for planning effective treatment. PMID- 19551852 TI - Elevated expression of microRNAs 155, 203, 210 and 222 in pancreatic tumors is associated with poorer survival. AB - Pancreatic cancer is the eighth most common cancer and has an overall 5-year survival rate lower than 10%. Because of their ability to regulate gene expression, microRNAs can act as oncogenes or tumor-suppressor genes and so have garnered interest as possible prognostic and therapeutic markers during the last decade. However, the prognostic value of microRNA expression in pancreatic cancer has not been thoroughly investigated. We measured the levels of miR-155, miR-203, miR-210, miR-216, miR-217 and miR-222 by quantitative RT-PCR in a cohort of 56 microdissected pancreatic ductal adenocarcinomas (PDAC). These microRNAs were chosen as they had previously been shown to be differentially expressed in pancreatic tumors compared to normal tissues. The possible association of microRNA expression and patients' survival was examined using multivariate Cox's regression hazard analyses. Interestingly, significant correlations between elevated microRNA expression and overall survival were observed for miR-155 (RR = 2.50; p = 0.005), miR-203 (RR = 2.21; p = 0.017), miR-210 (RR = 2.48; p = 0.005) and miR-222 (RR = 2.05; p = 0.035). Furthermore, tumors from patients demonstrating elevated expression levels of all 4 microRNAs possessed a 6.2-fold increased risk of tumor-related death compared to patients whose tumors showed a lower expression of these microRNAs. This study provides the first evidence for an oncogenic activity of miR-155, miR-203, miR-210 and miR-222 in the development of pancreatic cancer as has been reported for other tumor types. Furthermore, the putative target genes for these microRNAs suggest a complex signaling network that can affect PDAC tumorigenesis and tumor progression. PMID- 19551853 TI - Methylation of cystatin M promoter is associated with unfavorable prognosis in operable breast cancer. AB - The methylation status of cystatin M (CST6) gene in breast tumors was investigated and its prognostic significance as a novel breast cancer biomarker was evaluated. Using methylation-specific PCR (MSP), CST6 promoter methylation was examined in 134 formalin fixed paraffin-embedded tissues (FFPEs): 10 pairs of breast tumors and their surrounding normal tissues, 10 breast fibroadenomas, 11 normal breast tissues and 93 breast tumors. Methylation of CST6 promoter was observed in 2/21 (9.5%) noncancerous breast tissues, 1/10 (10%) benign breast tumors (fibroadenomas) and 52 (55.9%) operable breast cancer tumor samples. CST6 was rarely methylated in the normal tissue surrounding the tumor (10%). During the follow-up period, 24 (25.8%) patients relapsed and 19 (20.4%) died. CST6 methylation was detected in 19 (79.2%) of patients who relapsed and in 15 (78.9%) of patients who died. Disease-free-interval (DFI) and overall survival (OS) were significantly associated with CST6 promoter methylation (p=0.004 and p=0.001 respectively). Multivariate analysis revealed that CST6 methylation is an independent prognostic factor for DFI (HR=3.484; 95% CI: 1.155-10.511; p=0.027). and OS (HR=9.190; 95% CI: 1.989-42.454; p=0.004). CST6 promoter methylation status in tumor cells seems to provide important prognostic information in operable breast cancer and merits to be further evaluated and validated in a larger cohort of patients. PMID- 19551854 TI - Reproductive factors and postmenopausal hormone use in relation to endometrial cancer risk in the Nurses' Health Study cohort 1976-2004. AB - Endometrial cancer is a disease primarily driven by cumulative exposure to estrogen unopposed by progesterone. Reproductive factors associated with changes in endogenous hormone levels and use of exogenous hormones such as postmenopausal hormones influence the risk of disease. The authors used the Nurses' Health Study, comprised of 121,700 nurses, to assess the above associations. Over 28 years of follow-up, 778 adenocarcinoma cases were diagnosed and 1,850,078 person years were accumulated. Cox proportional hazards models were used to estimate relative risks (RR) and 95% confidence intervals (CI). A late age at menarche decreased the risk independent of body mass index (BMI) (P-trend = 0.02). A late age at menopause increased cancer risk (P-trend = 0.0003). An advanced age at last birth reduced the risk (P-trend < 0.0001), however, an inverse association with age at first birth and parity diminished after adjustment for age at last birth. Compared with never users, an increased risk was observed among long-term (> or =5 years) users of both estrogen (E) (RR = 7.67, 95% CI: 5.57, 10.57) and combined estrogen plus progesterone (E+P) (RR = 1.52, 95% CI: 1.03, 2.23). Normal weight (BMI < 25) women had the highest risk following E or E+P use (P interaction-E = 0.0008, P-interaction-E+P = 0.02). The findings from this study underscore the importance of hormonal mechanisms in endometrial carcinogenesis. PMID- 19551855 TI - Oncogenic human papillomaviruses block expression of the B-cell translocation gene-2 tumor suppressor gene. AB - Human papillomavirus (HPV)-induced carcinogenesis is critically dependent on the activities of the viral E6 and E7 oncogenes. Here, we demonstrate that expression of the putative tumor suppressor gene B-cell translocation gene-2 (BTG2) is reinduced in HPV16- and HPV18-positive cancer cells on silencing of viral oncogene expression, indicating that BTG2 is repressed by oncogenic HPVs. Inhibition of BTG2 expression was mediated by the HPV E6 oncogene and occurred in a p53-dependent manner. Luciferase reporter gene analyses revealed that BTG2 repression takes place at the transcriptional level and is dependent on the integrity of the major p53-response element within the BTG2 promoter. Ectopic expression of BTG2 acted antiproliferative in cervical cancer cells. Tissue specimens commonly exhibited reduced BTG2 protein levels in HPV-positive high grade lesions (CIN2/3) and cervical carcinomas, when compared with normal cervical epithelium. These findings identify the antiproliferative BTG2 gene as a novel cellular target blocked by the HPV E6 oncoprotein. PMID- 19551856 TI - Active MMP-2 effectively identifies the presence of colorectal cancer. AB - Fully active MMP-2 is expressed at such low levels in human tissues that studies often fail to confirm its value as a cancer marker despite strong associations with malignancy. Our study utilized careful extraction, accurate activity measurements, standardization to purified controls and a new statistical metric to determine whether active MMP-2 is an effective indicator of colorectal cancer compared to pro-MMP-2 or pro-MMP-9. MMP-2 and MMP-9 activities were analyzed in matched normal and cancer samples from 269 patients by gelatin zymography, computer-assisted image analysis, serial dilutions of strong samples and standardization to controls. An index of effect size was designed for comparative evaluation of active MMP-2, pro-MMP-2 and pro-MMP-9 activities. For each gelatinase, mean activity and protein levels/mg soluble protein in normal mucosa and colorectal cancer were calculated for the first time with respect to commercial standards. Active MMP-2 activity, detected in 99% of colorectal cancers, was higher in 95% of cancers (on average 10-fold) than in normal mucosa. Levels of pro-MMP-2 and pro-MMP-9, but not active MMP-9, activities were also significantly higher in cancers versus normal. However, active MMP-2 activity provided the most effective test for the presence of cancer (p<0.0.0001) with an effect size statistically significantly larger than for either pro-MMP-2 or pro MMP-9. Receiver operating characteristic (ROC) curves demonstrated that a cut-off for active MMP-2 of >44 SDU activity/mg soluble protein (>180 pg/mg), which is three times mean normal levels, would permit detection of colorectal cancer with an estimated sensitivity of 84% and estimated specificity of 93%. PMID- 19551857 TI - The mechanisms underlying MMR deficiency in immunodeficiency-related non-Hodgkin lymphomas are different from those in other sporadic microsatellite instable neoplasms. AB - The spectrum of tumors showing microsatellite instability (MSI) has recently been enlarged to sporadic neoplasms whose incidence is favored in the context of chronic immunosuppression. We investigated the biological, therapeutic and clinical features associated with MSI in immunodeficiency-related non-Hodgkin lymphomas (ID-RL). MSI screening was performed in 275 ID-RL. MSI ID-RL were further analyzed for MMR gene expression and for BRAF/KRAS mutations since these genes are frequently altered in MSI cancers. We also assessed the expression of O(6)-methylguanine-DNA methyltransferase (MGMT), an enzyme whose inactivation has been reported in lymphomas and may help in the selection of MMR deficient clones. Unlike other sporadic MSI neoplasms, MSI ID-RL (N = 17) presented with heterogeneous MMR defects and no MLH1 promoter methylation. About one third of these tumors presented with normal expression of MLH1, MSH2, MSH6 and PMS2. They accumulated BRAF activating mutations (33%). Unlike other ID-RL, MSI ID-RL were primarily EBV-negative NHL of T-cell origin, and arose after long-term immunosuppression in patients who received azathioprine as part of their immunosuppressive regimen (p = 0.05) and/or who exhibited methylation-induced loss of expression of MGMT in tumor cells (p= 0.02). Overall, these results highlight that, in the context of deficient immune status, some MSI neoplasms arise through alternative mechanism when compared to other sporadic MSI neoplasms. They give the exact way how to make the diagnosis of MSI in these tumors and may help to define biological and clinicalrisk factors associated with their emergence in such a clinicalcontext. PMID- 19551858 TI - Housekeeping genes in prostate tumorigenesis. AB - Housekeeping (HK) genes are involved in basic cellular functions and tend to be constitutively expressed across various tissues and conditions. A number of studies have analyzed the value of HK genes as an internal standard for assessing gene expression, but the role of HK genes in cancer development has never been specifically addressed. In this study, we sought to evaluate the expression of HK genes during prostate tumorigenesis. We performed a meta-analysis of gene expression during the transition from normal prostate (NP) to localized prostate cancer (LPC) (i.e., NP > LPC) and from localized to metastatic prostate cancer (MPC) (i.e., LPC > MPC). We found that HK genes are more likely to be differentially expressed during prostate tumorigenesis than is the average gene in the human genome, suggesting that prostate tumorigenesis is driven by modulation of the expression of HK genes. Cell-cycle genes and proliferation markers were up-regulated in both NP > LPC and LPC > MPC transitions. We also found that the genes encoding ribosomal proteins were up-regulated in the NP > LPC and down-regulated in the LPC > MPC transition. The expression of heat shock proteins was up-regulated during the LPC > MPC transition, suggesting that in its advanced stages, prostate tumor is under cellular stress. The results of these analyses suggest that during prostate tumorigenesis, there is a period when the tumor is under cellular stress and, therefore, may be the most vulnerable and responsive to treatment. PMID- 19551859 TI - Cancer risk among patients with cystic fibrosis and their first-degree relatives. AB - Patients with cystic fibrosis (CF) are at increased risk of some cancers. Little is known about the cancer risks among carriers heterozygous for the CF mutation and it is hypothesized this may be associated with reduced cancer risk. Using Swedish general population-based registers, we identified 884 patients with CF from 1968 to 2003 and 3,033 of their first-degree relatives The subjects were followed from birth of index persons or 1958, whichever came later, until death, emigration or 2003, whichever came first. Cancer risks were compared with the general Swedish population using standardized incidence ratios (SIR) with 95% confidence intervals (CI). Patients, followed for an average of 21 years, were at a higher overall risk of cancer. Some 26 cancer diagnoses, after excluding multiple diagnoses of nonmelanoma skin cancer in one man, produced an overall SIR of 3.2 (95% CI 2.1-4.6). We found statistically significantly increased risks for kidney, thyroid, endocrine, lymphoma and nonmelanoma skin cancer. There was no modification of cancer risk among parents and siblings, with an average of 21 years of follow-up. This study did not identify a heterozygote advantage for CF gene mutations in relation to cancer risk. PMID- 19551860 TI - Common germline polymorphisms in COMT, CYP19A1, ESR1, PGR, SULT1E1 and STS and survival after a diagnosis of breast cancer. AB - Although preliminary evidence suggests that germline variation in genes involved in steroid hormone metabolism may alter breast cancer prognosis, this has not been systematically evaluated. We examined associations between germline polymorphisms in 6 genes involved in the steroid hormone metabolism and signaling pathway (COMT, CYP19A1, ESR1, PGR, SULT1E1, STS) and survival among women with breast cancer participating in SEARCH, a population-based case-control study. Blood samples from up to 4,470 women were genotyped for 4 possible functional SNPs in CYP19A1 and 106 SNPs tagging the common variation in the remainder of the genes. The genotypes of each polymorphism were tested for association with survival after breast cancer diagnosis using Cox regression analysis. Significant evidence of an association was observed for a COMT polymorphism (rs4818 p = 0.016) under the codominant model. This SNP appeared to fit a dominant model better (HR = 0.80 95% CI: 0.69-0.95, p = 0.009); however, the result was only marginally significant after permutation analysis adjustment for multiple hypothesis tests (p = 0.047). To further evaluate this finding, somatic expression microarray data from 8 publicly available datasets were used to test the association between survival and tumor COMT gene expression; no statistically significant associations were observed. A correlated SNP in COMT, rs4860, has recently been associated with breast cancer prognosis in Chinese women in a dominant model. These results suggest that COMT rs4818, or a variant it tags, is associated with breast cancer prognosis. Further study of COMT and its putative association with breast cancer prognosis is warranted. PMID- 19551861 TI - Metabolic regulation and redox activity as mechanisms for angioprevention by dietary phytochemicals. AB - The existence of active principles in numerous foods and beverages has been recognized by traditional medicines worldwide after centuries of empirical trial. Epidemiological studies support the concepts linking diet to survival, particularly in the incidence rates of specific cancers. Molecular studies have provided evidence that a wide range of food-derived phytochemicals and other diet associated compounds or their synthetic derivatives represent a cornucopia of potential new compounds for prevention and treatment of chronic or acute diseases. Many have entered clinical practice or are under clinical testing. A remarkable property shared by several phytochemicals is the capacity to restrain inflammation and angiogenesis, two complex physiologic processes kept under control by strict rules, which can backfire in cancer and in pathologic conditions such as metabolic, cardiovascular and neurological disorders. We termed this concept "angioprevention". Here, we discuss recent findings on the metabolic effects of several phytochemicals with anticancer properties. The different molecular targets shared by these compounds seem to converge on crosstalking signaling networks involved in controlling energy metabolism through a redox-regulated code. The redox imbalance produced in the tissue microenvironment elicits an adaptive response that seems to provide cytoprotective effects potentially beneficial in cardiovascular and neurological disorders or energy balancing effects in metabolic disorders. However, in transformed and overt tumor cells, this redox imbalance favors cell death while curbing tumor inflammation and angiogenesis, thus engaging an overall antitumor response. These concepts provide a broader framework for pharmacological application of phytochemical-derived drugs against cancer. PMID- 19551862 TI - Merkel cell carcinoma subgroups by Merkel cell polyomavirus DNA relative abundance and oncogene expression. AB - Merkel cell polyomavirus (MCPyV) was recently discovered in Merkel cell carcinoma (MCC), a clinically and pathologically heterogeneous malignancy of dermal neuroendocrine cells. To investigate this heterogeneity, we developed a tissue microarray (TMA) to characterize immunohistochemical staining of candidate tumor cell proteins and a quantitative PCR assay to detect MCPyV and measure viral loads. MCPyV was detected in 19 of 23 (74%) primary MCC tumors, but 8 of these had less than 1 viral copy per 300 cells. Viral abundance of 0.06-1.2 viral copies/cell was directly related to presence of retinoblastoma gene product (pRb) and terminal deoxyribonucleotidyl transferase (TdT) by immunohistochemical staining (p < or = 0.003). Higher viral abundance tumors tended to be associated with less p53 expression, younger age at diagnosis and longer survival (p < or = 0.08). These data suggest that MCC may arise through different oncogenic pathways, including ones independent of pRb and MCPyV. PMID- 19551863 TI - Id-1 promotes tumorigenicity and metastasis of human esophageal cancer cells through activation of PI3K/AKT signaling pathway. AB - Id-1 (inhibitor of differentiation or DNA binding) is a helix-loop-helix protein that is overexpressed in many types of cancer including esophageal squamous cell carcinoma (ESCC). We previously reported that ectopic Id-1 expression activates the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway in human esophageal cancer cells. In this study, we confirmed a positive correlation between Id-1 and phospho-AKT (Ser473) expressions in ESCC cell lines, as well as in ESCC on a tissue microarray. To investigate the significance of Id 1 in esophageal cancer progression, ESCC cells with stable ectopic Id-1 expression were inoculated subcutaneously into the flank of nude mice and were found to form larger tumors that showed elevated Ki-67 proliferation index and increased angiogenesis, as well as reduced apoptosis, compared with control cells expressing the empty vector.The Id-1-overexpressing cells also exhibited enhanced metastatic potential in the experimental metastasis assay. Treatment with the PI3K inhibitor LY294002 attenuated the tumor promotion effects of Id-1, indicating that the effects were mediated by the PI3K/AKT signaling pathway. In addition, our in vitro experiments showed that ectopic Id-1 expression altered the expression levels of markers associated with epithelial-mesenchymal transition and enhanced the migration ability of esophageal cancer cells. The Id 1-overexpressing ESCC cells also exhibited increased invasive potential, which was in part due to PI3K/AKT-dependent modulation of matrix metalloproteinase-9 expression. In conclusion, our results provide the first evidence that Id-1 promotes tumorigenicity and metastasis of human esophageal cancer in vivo and that the PI3K inhibitor LY294002 can attenuate these effects. PMID- 19551864 TI - Genetic variation and circulating levels of IGF-I and IGFBP-3 in relation to risk of proliferative benign breast disease. AB - Insulin-like growth factor-I (IGF-I) and its major binding protein IGFBP-3 have been implicated in breast carcinogenesis. We examined the associations between genetic variants and circulating levels of IGF-I and IGFBP-3 with proliferative benign breast disease (BBD), a marker of increased breast cancer risk, in the Nurses' Health Study II (NHSII). Participants were 359 pathology-confirmed proliferative BBD cases and 359 matched controls. Circulating IGF-I and IGFBP-3 levels were measured in blood samples collected between 1996 and 1999. Thirty single nucleotide polymorphisms (SNPs) in IGF-I, IGFBP-1, and IGFBP-3 genes were selected using a haplotype tagging approach and genotyped in cases and controls. Circulating IGF-I levels were not associated with proliferative BBD risk. Higher circulating IGFBP-3 levels were significantly associated with increased risk of proliferative BBD (highest vs. lowest quartile odds ratio (OR) [95% confidence interval (CI)], 1.70 (1.06-2.72); p-trend = 0.03). The minor alleles of 2 IGFBP-3 SNPs were associated with lower proliferative BBD risk (homozygous variant vs. homozygous wild-type OR (95% CI): rs3110697: 0.6 (0.4-0.9), p-trend = 0.02; rs2132570: 0.2 (0.1-0.6), p-trend = 0.02). Three other IGFBP-3 SNPs (rs2854744, rs2960436 and rs2854746) were significantly associated with circulating IGFBP-3 levels (p < 0.01). Although these SNPs were not significantly associated with proliferative BBD risk, there was suggestive evidence that the alleles associated with higher circulating IGFBP-3 levels were also associated with higher risk of proliferative BBD. These results suggest that genetic variants and circulating levels of IGFBP-3 may play a role in the early stage of breast carcinogenesis. PMID- 19551865 TI - The YSNSG cyclopeptide derived from tumstatin inhibits tumor angiogenesis by down regulating endothelial cell migration. AB - We previously demonstrated that the CNYYSNS peptide derived from tumstatin inhibited in vivo tumor progression. The YSNS motif formed a beta-turn crucial for biological activity. More recently, a YSNSG cyclopeptide with a constrained beta-turn on the YSNS residues was designed. Intraperitoneal administration of the YSNSG cyclopeptide inhibited in vivo melanoma progression more efficiently than the native linear peptide. In the present article, we showed that the YSNSG cyclopeptide also triggered an inhibition of in vivo tumor neovascularization and we further analyzed its in vitroantiangiogenic effect. The YSNSG cyclopeptide did not alter endothelial cell proliferation but inhibited cell migration by 83% in an in vitro wound healing model. The inhibition was mediated by a decrease in active MT1-MMP at the migration front as well as a decrease in u-PA and u-PAR expression. The cyclopeptide also altered beta1-integrin distribution in endothelial cell lamellipodia, induced a strong decrease in the phosphorylated focal adhesion kinase (p125(FAK)), disorganized F-actin stress fibers and decreased the number of lamellipodia, resulting in a non migratory phenotype. Our results confirm the YSNSG cyclopeptide as a potent antitumor agent, through both the inhibition of invasive properties of tumor cells and the antiangiogenic activity. PMID- 19551866 TI - N-glycosylation status of beta-haptoglobin in sera of patients with colon cancer, chronic inflammatory diseases and normal subjects. AB - N-glycosylation status of purified beta-haptoglobin from sera of 17 patients, and from sera of 14 healthy volunteer subjects, was compared by blotting with various lectins and antibodies. Patients in this study were diagnosed as having colon cancer through histological examination of each tumor tissue by biopsy. Blotting index of serum beta-haptoglobin with Aleuria aurantia lectin (AAL) was clearly higher for cancer patients than for healthy subjects. No such distinction was observed for blotting with three other lectins and two monoclonal antibodies. To determine tumor-associated reactivity of AAL binding as compared to inflammatory processes in colonic tissues, beta-haptoglobin separated from sera of 5 patients with Crohn's disease (CD), and 4 patients with ulcerative colitis (UC), was studied. All these cases, except one case of UC, showed AAL index lower than that in cancer cases, similarly to healthy subjects. The higher AAL binding of beta haptoglobin in colon cancer patients than in healthy subjects appeared to be due to alpha-L-fucosyl residue, since it was eliminated by bovine kidney alpha fucosidase treatment. N-linked glycans of serum haptoglobin from colon cancer patients vs. healthy subjects were released by N-glycanase, fluorescence-labeled, and subjected to normal-phase high performance liquid chromatography (NP-HPLC). Glycan structures were determined based on glucose unit (GU) values and their changes upon sequential treatment with various exoglycosidases. Glycosyl sequences and their branching status of glycans from 14 cases of serum beta haptoglobin were characterized. The identified glycans were sialylated or nonsialylated, bi-antennary or tri-antennary structures, with or without terminal fucosylation. PMID- 19551867 TI - BRCA1 modulates malignant cell behavior, the expression of survivin and chemosensitivity in human breast cancer cells. AB - BRCA1 is a multifunctional tumor-suppressive protein. Many functional aspects of BRCA1 are not fully understood. We used a shRNA approach to probe the function of BRCA1 in human breast cancer cells. Knocking down BRCA1 expression by shRNA in the wild-type BRCA1 human breast cancer MCF-7 and MDA-MB-231 cells resulted in an increase in cell proliferation, anchorage-independent growth, cell migration, invasion and a loss of p21/Waf1 and p27Kip1 expression. In BRCA1 knocked-down cells, the expression of survivin was significantly up regulated with a concurrent decrease in cellular sensitivity to paclitaxel. We also found that cells harboring endogenous mutant or defective BRCA1 (MDA-MB-436 and HCC1937) were highly proliferative and expressed a relatively low level of p21/Waf1 and p27Kip1 by comparison to wild-type BRCA1 cells. Cells harboring mutated BRCA1 also expressed a high level of survivin and were relatively resistant to paclitaxel by comparison to wild-type cells. Increase resistance to paclitaxel was due to an increase in the expression of survivin in both the BRCA1 knocked down and mutant BRCA1 cells because knocking down survivin expression by siRNA restored sensitivity to paclitaxel. We conclude that BRCA1 down-modulates the malignant behavior of breast cancer cells, promotes the expression of p21/Waf1, p27Kip1 and inhibits the expression of survivin. Moreover, loss of BRCA1 expression or function leads to an increase in survivin expression and a reduction in chemosensitivity to paclitaxel. PMID- 19551869 TI - Hyperossification in miniaturized toadlets of the genus Brachycephalus (Amphibia: Anura: Brachycephalidae): Microscopic structure and macroscopic patterns of variation. AB - Species of the genus Brachycephalus, have a snout-vent length of less than 18 mm and are believed to have evolved through miniaturization. Brachycephalus ephippium, is particularly interesting; because its entire skull is hyperossified, and the presacral vertebrae and transverse processes are covered by a dorsal shield. We demonstrate in this paper that, at the macroscopic level, a completely hyperossified skull and dorsal shield occur only in B. ephippium, but not in B. ferruginus, B. izechsohni, B. pernix, B. pombali, B. brunneus, B. didactylus, and B. hermogenesi. An intermediate condition, in which the skull is hyperossified but a dorsal shield is absent, occurs in B. vertebralis, B. nodoterga, B. pitanga, and B. alipioi. The microscopic structure of hyperossification was examined in skulls of B. ephippium and B. pitanga, revealing a complex organization involving the presence of Sharpey fibers, which in humans are characteristic of periodontal connections. PMID- 19551868 TI - KLF4-dependent, PPARgamma-induced expression of GPA33 in colon cancer cell lines. AB - The glycoprotein A33 (GPA33) is a colon cancer antigen. Phase I trials with 131I and 125I monoclonal antibody A33 in colon carcinoma patients showed excellent localization to colorectal cancer and some evidence of tumor response. Using DNA microarrays, we have identified the GPA33 gene as a target of PPARgamma in HT29 Cl.16E colon cancer cells. Treatment of HT29-Cl.16E, Caco2, SW1116 and LS174T colon cancer cells with the PPARgamma agonist GW7845 induced a 2- to 6-fold increase in GPA33 mRNA as determined by real-time PCR. This induction was also found in HT29-Cl.16E cells treated with rosiglitazone and ciglitazone and was prevented by cotreatment with the PPARgamma antagonist GW9662, indicating that this regulation was PPARgamma dependent. No canonical PPAR responsive element was found in the GPA33 promoter. We therefore analyzed the expression of transcription factors involved in GPA33 expression. CDXl, CDX2 and KLF5 expression was not modified by PPARgamma activation. By contrast, a significant increase in KLF4 was seen, both at mRNA and protein levels. Furthermore, chromatin immunoprecipitation studies demonstrated that an increased amount of KLF4 protein was bound to the GPA33 promoter in cells treated with rosiglitazone. Finally, downregulation of KLF4 expression by siRNA reduced rosiglitazone-induced GPA33 expression. This indicates that PPARgamma activation induces KLF4 expression, which in turn increases GPA33 expression. We also demonstrate that PPARgamma activation leads to increased (p21WAF1/Cip1 and keratin 19) or decreased (cyclin D1) expression of known KLF4 targets, suggesting that KLF4 is a nodal player in a network of PPARgamma-regulated genes. PMID- 19551870 TI - The lissamphibian humerus and elbow joint, and the origins of modern amphibians. AB - The origins and evolution of the three major clades of modern amphibians are still a source of controversy, and no general consensus exists as to their relationship to the various known Paleozoic taxa. This may indicate that additional character complexes should be studied to resolve their phylogenetic relationship. The salamander elbow joint has been fundamentally misinterpreted in previous morphological descriptions. In caudates and anurans, both the radius and ulna (fused in anurans) articulate with the characteristically large capitulum (radial condyle), although part of the ulnar articulating surface fits into to the smooth trochlear region. The salamander "ulnar condyle" of previous descriptions is in fact the entepicondyle. The condition seen in batrachians (i.e., salamanders and frogs) may be a lissamphibian synapomorphy because the elbow region of the primitive fossil caecilian Eocaecilia resembles those of frogs and salamanders. In addition to the large and bulbous capitulum, all lissamphibian humeri lack an entepicondylar foramen, and possess a distally pointing entepicondyle, a low and rounded ectepicondyle, and an elongated shaft. These characters are identified in key fossil forms to assess the support for the different hypotheses proposed for the evolutionary origins of lissamphibians. Temnospondyli is the only group of early tetrapods that shows a progressive evolution of lissamphibian traits in the humerus and elbow joint. Furthermore, among Paleozoic taxa, the dissorophoid temnospondyl Doleserpeton annectens is the only taxon that has the full set of humeral features shared by all lissamphibians. These results add support for the theory of a monophyletic origin of lissamphibians from dissorophoidtemnospondyls. PMID- 19551871 TI - Histology and ultrastructure of the placental membranes of the viviparous brown snake, Storeria dekayi (Colubridae: Natricinae). AB - The placental membranes of the viviparous brown snake Storeria dekayi were examined following mid-gestation by means of light microscopy, scanning electron microscopy, and transmission electron microscopy to reveal their structural organization and cytological composition. By Zehr stage 32, the chorioallantoic placenta (allantoplacenta) is established around much of the egg, and a well developed omphalallantoic placenta occurs in the abembryonic hemisphere. The allantoplacenta exhibits multiple features that enhance interhemal exchange: the uterus and allantois are well vascularized, the chorionic and uterine epithelia are attenuated, and the shell membrane is vestigial and has begun to degenerate. In the omphalallantoic placenta, the uterine epithelium is enlarged and appears to be secretory. The omphalopleure contains two distinct populations of cells, and shows cytological evidence for absorption. In intermediate areas, regions of omphalallantoic placenta are being transformed into allantoplacenta, through depletion of the isolated yolk mass and reduction in epithelial height of both uterus and omphalopleure. Morphological evidence suggests that the allantoplacenta is specialized for gas exchange, and the omphalallantoic placenta, for maternal secretion and fetal absorption. On the basis of the available evidence, we postulate that this pattern is characteristic of the thamnophine radiation of snakes. PMID- 19551874 TI - Differential expression of TrkB isoforms switches climbing fiber-Purkinje cell synaptogenesis to selective synapse elimination. AB - Correct neural function depends on precisely organized connectivity, which is refined from broader projections through synaptic/collateral elimination. In the rat, olivocerebellar topography is refined by regression of multiple climbing fiber (CF) innervation of Purkinje cells (PC) during the first two postnatal weeks. The molecules that initiate this regression are not fully understood. We assessed the role of cerebellar neurotrophins by examining tropomycin receptor kinase (Trk) receptor expression in the inferior olive and cerebellum between postnatal days (P)3-7, when CF-PC innervation changes from synapse formation to selective synapse elimination, and in a denervation-reinnervation model when synaptogenesis is delayed. Trks A, B, and C are expressed in olivary neurons; although TrkA was not transported to the cerebellum and TrkC was unchanged during innervation and reinnervation, suggesting that neither receptor is involved in CF PC synaptogenesis. In contrast, both total and truncated TrkB (TrkB.T) increased in the olive and cerebellum from P4, whereas full-length and activated phosphorylated TrkB (phospho-TrkB) decreased from P4-5. This reveals less TrkB signaling at the onset of CF regression. This expression pattern was reproduced during CF-PC reinnervation: in the denervated hemicerebellum phospho-TrkB decreased as CF terminals degenerated, then increased in parallel with the delayed neosynaptogenesis as new CFs reinnervated the denervated hemicerebellum. In the absence of this signaling, CF reinnervation did not develop. Our data reveal that olivocerebellar TrkB activity parallels CF-PC synaptic formation and stabilization and is required for neosynaptogenesis. Furthermore, TrkB.T expression rises to reduce TrkB signaling and permit synapse elimination. PMID- 19551875 TI - Error propagation from prime variables into specific rates and metabolic fluxes for mammalian cells in perfusion culture. AB - Error propagation from prime variables into specific rates and metabolic fluxes was quantified for high-concentration CHO cell perfusion cultivation. Prime variable errors were first determined from repeated measurements and ranged from 4.8 to 12.2%. Errors in nutrient uptake and metabolite/product formation rates for 5-15% error in prime variables ranged from 8-22%. The specific growth rate, however, was characterized by higher uncertainty as 15% errors in the bioreactor and harvest cell concentration resulted in 37.8% error. Metabolic fluxes were estimated for 12 experimental conditions, each of 10 day duration, during 120-day perfusion cultivation and were used to determine error propagation from specific rates into metabolic fluxes. Errors of the greater metabolic fluxes (those related to glycolysis, lactate production, TCA cycle and oxidative phosphorylation) were similar in magnitude to those of the related greater specific rates (glucose, lactate, oxygen and CO(2) rates) and were insensitive to errors of the lesser specific rates (amino acid catabolism and biosynthesis rates). Errors of the lesser metabolic fluxes (those related to amino acid metabolism), however, were extremely sensitive to errors of the greater specific rates to the extent that they were no longer representative of cellular metabolism and were much less affected by errors in the lesser specific rates. We show that the relationship between specific rate and metabolic flux error could be accurately described by normalized sensitivity coefficients, which were readily calculated once metabolic fluxes were estimated. Their ease of calculation, along with their ability to accurately describe the specific rate metabolic flux error relationship, makes them a necessary component of metabolic flux analysis. PMID- 19551873 TI - Rapid, long-term labeling of cells in the developing and adult rodent visual cortex using double-stranded adeno-associated viral vectors. AB - Chronic in vivo imaging studies of the brain require a labeling method that is fast, long-lasting, efficient, nontoxic, and cell-type specific. Over the last decade, adeno-associated virus (AAV) has been used to stably express fluorescent proteins in neurons in vivo. However, AAV's main limitation for many studies (such as those of neuronal development) is the necessity of second-strand DNA synthesis, which delays peak transgene expression. The development of double stranded AAV (dsAAV) vectors has overcome this limitation, allowing rapid transgene expression. Here, we have injected different serotypes (1, 2, 6, 7, 8, and 9) of a dsAAV vector carrying the green fluorescent protein (GFP) gene into the developing and adult mouse visual cortex and characterized its expression. We observed labeling of both neurons and astrocytes with serotype-specific tropism. dsAAV-GFP labeling showed high levels of neuronal GFP expression as early as 2 days postinjection and as long as a month, surpassing conventional AAV's onset of expression and matching its longevity. Neurons labeled with dsAAV-GFP appeared structurally and electrophysiologically identical to nonlabeled neurons, suggesting that dsAAV-GFP is neither cytotoxic nor alters normal neuronal function. We also demonstrated that dsAAV-labeled cells can be imaged with subcellular resolution in vivo over multiple days. We conclude that dsAAV is an excellent vector for rapid labeling and long-term in vivo imaging studies of astrocytes and neurons on the single cell level within the developing and adult visual cortex. PMID- 19551876 TI - Synergistic coordination of polyethylene glycol with ClpB/DnaKJE bichaperone for refolding of heat-denatured malate dehydrogenase. AB - The use of polyethylene glycol (PEG) as a refolding additive to a refolding cocktail comprising the molecular bichaperone ClpB and DnaKJE significantly enhances chaperone-mediated refolding of heat-denatured malate dehydrogenase (MDH). The critical factor to affect the refolding yield is the time point of introducing PEG to the refolding cocktail. The refolding efficiency reached approximately 90% only when PEG was added at the beginning of refolding reaction. The synergistic coordination of an inexpensive refolding additive PEG with the ClpB/DnaKJE bichaperone system may provide an economical route to further enhance the efficacy of ClpB/DnaKJE refolding cocktail approach, facilitating its implementation in large-scale refolding processes. PMID- 19551878 TI - Chiral separation of phosphine-containing alpha-amino acid derivatives using two complementary cellulosic stationary phases in supercritical fluid chromatography. AB - Enantiomeric separations of six amino-acid derivatives have been studied using packed-column supercritical fluid chromatography with two polysaccharide-based enantioselective stationary phases: cellulose tris(3,5-dimethylphenylcarbamate) and cellulose tris(3-chloro-4-methylphenylcarbamate) (Lux Cellulose-1 and -2). The effect of analyte structure on retention and separation was studied. Varied mobile phase compositions were investigated: alcohol modifier percentage was increased from 3 to 40% but smaller amounts were most effective in separating these compounds. Besides, ethanol was preferred to methanol or isopropanol as it proved to be a good compromise to achieve sufficient resolution in a reasonable analysis time. Moreover, a carbon dioxide-ethanol mixture allows performing analyses in safe and green conditions. The effect of temperature at constant mobile phase composition was explored between 10 and 40 degrees C. In most cases, increasing the temperature improved the chiral separation, up to an optimum temperature. The results are discussed in line with the structure variation of the racemic derivatives analyzed and the two columns are compared. The two columns were shown to provide complementary selectivities for the investigated solutes: whereas Lux 1 provided separation for five of the six racemates, Lux 2 could resolve the last racemic mixture. Finally, optimized conditions of separation are defined. PMID- 19551877 TI - Mcl-1 overexpression leads to higher viabilities and increased production of humanized monoclonal antibody in Chinese hamster ovary cells. AB - Bioreactor stresses, including nutrient deprivation, shear stress, and byproduct accumulation can cause apoptosis, leading to lower recombinant protein yields and increased costs in downstream processing. Although cell engineering strategies utilizing the overexpression of antiapoptotic Bcl-2 family proteins such as Bcl-2 and Bcl-x(L) potently inhibit apoptosis, no studies have examined the use of the Bcl-2 family protein, Mcl-1, in commercial mammalian cell culture processes. Here, we overexpress both the wild type Mcl-1 protein and a Mcl-1 mutant protein that is not degraded by the proteasome in a serum-free Chinese hamster ovary (CHO) cell line producing a therapeutic antibody. The expression of Mcl-1 led to increased viabilities in fed-batch culture, with cell lines expressing the Mcl-1 mutant maintaining approximately 90% viability after 14 days when compared with 65% for control cells. In addition to enhanced culture viability, Mcl-1 expressing cell lines were isolated that consistently showed increases in antibody production of 20-35% when compared with control cultures. The quality of the antibody product was not affected in the Mcl-1-expressing cell lines, and Mcl 1-expressing cells exhibited 3-fold lower caspase-3 activation when compared with the control cell lines. Altogether, the expression of Mcl-1 represents a promising alternative cell engineering strategy to delay apoptosis and increase recombinant protein production in CHO cells. PMID- 19551880 TI - The fermentation stoichiometry of Thermotoga neapolitana and influence of temperature, oxygen, and pH on hydrogen production. AB - The hyperthermophilic bacterium, Thermotoga neapolitana, has potential for use in biological hydrogen (H(2)) production. The objectives of this study were to (1) determine the fermentation stoichiometry of Thermotoga neapolitana and examine H(2) production at various growth temperatures, (2) investigate the effect of oxygen (O(2)) on H(2) production, and (3) determine the cause of glucose consumption inhibition. Batch fermentation experiments were conducted at temperatures of 60, 65, 70, 77, and 85 degrees C to determine product yield coefficients and volumetric productivity rates. Yield coefficients did not show significant changes with respect to growth temperature and the rate of H(2) production reached maximum levels in both the 77 degrees C and 85 degrees C experiments. The fermentation stoichiometry for T. neapolitana at 85 degrees C was 3.8 mol H(2), 2 mol CO(2), 1.8 mol acetate, and 0.1 mol lactate produced per mol of glucose consumed. Under microaerobic conditions H(2) production did not increase when compared to anaerobic conditions, which supports other evidence in the literature that T. neapolitana does not produce H(2) through microaerobic metabolism. Glucose consumption was inhibited by a decrease in pH. When pH was adjusted with buffer addition cultures completely consumed available glucose. PMID- 19551879 TI - Monomeric structure of the cardioprotective chemokine SDF-1/CXCL12. AB - The chemokine stromal cell-derived factor-1 (SDF-1/CXCL12) directs leukocyte migration, stem cell homing, and cancer metastasis through activation of CXCR4, which is also a coreceptor for T-tropic HIV-1. Recently, SDF-1 was shown to play a protective role after myocardial infarction, and the protein is a candidate for development of new anti-ischemic compounds. SDF-1 is monomeric at nanomolar concentrations but binding partners promote self-association at higher concentrations to form a typical CXC chemokine homodimer. Two NMR structures have been reported for the SDF-1 monomer, but only one matches the conformation observed in a series of dimeric crystal structures. In the other model, the C terminal helix is tilted at an angle incompatible with SDF-1 dimerization. Using a rat heart explant model for ischemia/reperfusion injury, we found that dimeric SDF-1 exerts no cardioprotective effect, suggesting that the active species is monomeric. To resolve the discrepancy between existing models, we solved the NMR structure of the SDF-1 monomer in different solution conditions. Irrespective of pH and buffer composition, the C-terminal helix remains tilted at an angle with no evidence for the perpendicular arrangement. Furthermore, we find that phospholipid bicelles promote dimerization that necessarily shifts the helix to the perpendicular orientation, yielding dipolar couplings that are incompatible with the NOE distance constraints. We conclude that interactions with the alignment medium biased the previous structure, masking flexibility in the helix position that may be essential for the distinct functional properties of the SDF 1 monomer. PMID- 19551881 TI - Cancer treatment adherence among low-income women with breast or gynecologic cancer: a randomized controlled trial of patient navigation. AB - BACKGROUND: The authors implemented a controlled, randomized trial that compared 2 interventions: the provision of written resource navigation information (enhanced usual care [EUC]) versus written information plus patient navigation (TPN) aimed at improving adjuvant treatment adherence and follow-up among 487 low income, predominantly Hispanic women with breast cancer or gynecologic cancer. METHODS: Women were randomized to receive either TPN or EUC; and chemotherapy, radiation therapy, hormone therapy, and follow-up were assessed over 12 months. Patients with breast cancer were analyzed separately from patients with gynecologic cancer. RESULTS: Overall adherence rates ranged from 87% to 94%, and there were no significant differences between the TPN group and the EUC group. Among women with breast cancer, 90% of the EUC group and 88% of the TPN group completed chemotherapy (14% of the EUC group and 26% of the TPN group delayed the completion of chemotherapy), 2% of the EUC group and 4% of the TPN group failed to complete chemotherapy, and 8% of the EUC group and 7% of the TPN group refused chemotherapy. Radiation treatment adherence was similar between the groups: Ninety percent of patients completed radiation (40% of the EUC group and 42% of the TPN group delayed the completion of radiation); in both groups, 2% failed to complete radiation, and 8% refused radiation. Among gynecologic patients, 87% of the EUC group and 94% of the TPN group completed chemotherapy (41% of the EUC group and 31% of the TPN group completed it with delays), 7% of the EUC group and 6% of the TPN group failed to complete chemotherapy, 6% of the EUC refused chemotherapy, 87% of the EUC group and 84% of the TPN group completed radiation (51% of the EUC group and 42% of the TPN with delays), 5% of the EUC group and 8% of the TPN group failed to complete radiation, and 8% of the EUC group and 5% of the TPN group refused radiation. CONCLUSIONS: Treatment adherence across randomized groups was notably higher than reported in previous studies, suggesting that active telephone patient navigation or written resource informational materials may facilitate adherence among low-income, predominantly Hispanic women. Adherence also may have be facilitated by federal-state breast and cervical cancer treatment funding. PMID- 19551882 TI - Etiology and outcome of extreme leukocytosis in 758 nonhematologic cancer patients: a retrospective, single-institution study. AB - BACKGROUND: To the authors' knowledge, the literature regarding extreme leukocytosis in solid tumor patients is sparse, consisting of a few case reports and small case series. METHODS: A total of 3770 consecutive solid tumor patients with a white blood cell count>40,000/microL were retrospectively identified over a 3-year period (2005-2008). Those patients without a secondary cause of their leukocytosis were defined as having a paraneoplastic leukemoid reaction. RESULTS: A total of 758 (20%) patients with solid tumors and extreme leukocytosis were identified. The etiology of the leukocytosis was hematopoietic growth factors in 522 (69%) patients, infection in 112 (15%) patients, high-dose corticosteroids in 38 (5%) patients, newly diagnosed leukemia in 9 (1%) patients, and paraneoplastic leukemoid reaction in 77 (10%) patients. The patients diagnosed with a paraneoplastic leukemoid reaction typically had neutrophil predominance (96%) and radiographic evidence of metastatic disease (78%), were clinically stable, and had a poor prognosis; 78% either died or were discharged to hospice within 12 weeks of their initial extreme leukocyte count. All of the 8 (10%) patients who survived>1 year received effective antineoplastic therapy. CONCLUSIONS: Infection was an uncommon cause of extreme leukocytosis in patients with solid tumors. Patients with paraneoplastic leukemoid reactions typically were clinically stable despite having large tumor burdens. However, clinical outcomes were poor unless effective antineoplastic treatment was received. PMID- 19551883 TI - Platin-based exclusive chemotherapy for selected patients with squamous cell carcinoma of the larynx and pharynx. AB - BACKGROUND: The current study was conducted to determine the long-term outcomes of patients with squamous cell carcinoma of the larynx and pharynx who were treated with platin-based exclusive chemotherapy (EC) after they achieved a complete clinical response (CCR) to induction chemotherapy. METHODS: One hundred forty-two who achieved a CCR after platin-based induction chemotherapy were treated exclusively with additional chemotherapy, and 98.6% were followed for a minimum of 3 years or until death. Thirty-five patients had >10 years of follow up. RESULTS: The survival rates at 1 year and 5 years were 95.8% and 61.2%, respectively. The main causes of death were metachronous second primary tumors (n = 27) and intercurrent disease (n = 21). Death related to EC was not encountered, and only 2 patients (1.4%) had grade 4 toxicity. In multivariate analysis, primary tumor arising outside the glottic larynx (P = .0001) and a Charlson comorbidity index >1 (P = .0001) were associated with a statistically significant reduction in survival. The 1-year and 5-year Kaplan-Meier local control estimates were 76.1% and 50.7%, respectively. Salvage treatment resulted in an observed final local control rate of 93% that varied from 97.2% in patients who had glottic cancer to 88.7% in patients who had tumor originating from other sites (P = .097). Combined chemotherapy with cisplatin and 5-fluorouracil (PF) allowed for the successful modulation of local therapy in 54.9% of patients. CONCLUSIONS: For selected patients, EC may provide long-term, durable disease control. For patients who developed recurrent disease after EC, this approach did not diminish survival and maintained function in the majority of patients. Future work should be directed toward select markers of response to PF chemotherapy with which to identify those patients who are suited optimally for this approach. PMID- 19551884 TI - Outcomes of patients with stage III nonsmall cell lung cancer treated with chemotherapy and radiation with and without surgery. AB - BACKGROUND: The objective of this study was to identify the factors associated with improved outcome after treatment for stage III nonsmall cell lung cancer (NSCLC). METHODS: A retrospective review of stage III NSCLC patients treated at who were treated at the Dana-Farber Cancer Institute/Brigham and Women's Cancer Center was done with institutional review board approval. Patients were followed for toxicity, local and distant failure, and overall survival. Multivariate Cox logistic regression analysis was used to determine the factors associated with treatment outcome. RESULTS: Between August 2000 and November 2006, 144 patients received concurrent chemoradiation (CRT) for stage III NSCLC. Eighty of 144 patients were men (56%), and the median age was 61 years (range, 33-81 years). Sixty-two patients (43%) had stage IIIA NSCLC, and 82 patients (57%) had stage IIIB NSCLC. Radiotherapy (RT) was given concurrently with chemotherapy to all patients; 100 patients (69%) received CRT without surgery, and 44 patients (31%) received with neoadjuvant CRT followed by surgical resection. The median RT dose was 60 grays (Gy) (range, 46-70 Gy). The median follow-up was 15 months (range, 3 64 months), the median potential follow-up was 37 months (range, 12-84 months), and the median overall survival was 22 months (95% confidence interval, 15-28 months). The 1-year and 2-year survival rates were 68% and 47%, respectively. Among the 44 patients who underwent resection, the median survival was 61 months, and the 2-year survival rate was 73%. On multivariate analysis, stage at the time of treatment (stage IIIA vs stage IIIB) and use of surgery were the only factors associated with improved outcome (P=.01 and P=.001, respectively). CONCLUSIONS: In this retrospective series, those patients who were able to undergo resection appeared to have improved outcome after induction CRT. PMID- 19551885 TI - DFT/TD-DFT investigation on Ir(III) complexes with N-heterocyclic carbene ligands: geometries, electronic structures, absorption, and phosphorescence properties. AB - Iridium(III) complexes with N-heterocyclic (NHC) ligands including fac-Ir(pmb)(3) (1), mer-Ir(pmb)(3) (2), (pmb)(2)Ir(acac) (3), mer-Ir(pypi)(3) (4), and fac Ir(pypi)(3) (5) [pmb = 1-phenyl-3H-benzimidazolin-2-ylidene, acac = acetoylacetonate, pypi = 1-phenyl-5H-benzimidazolin-2-ylidene; fac = facial, mer = meridional] were investigated theoretically. The geometry structures of 1-5 in the ground and excited state were optimized with restricted and unrestricted DFT (density functional theory) methods, respectively (LANL2DZ for Ir atom and 6-31G for other atoms). The HOMOs (highest occupied molecular orbitals) of 1-3 are composed of d(Ir) and pi(phenyl), while those of 4 and 5 are contributed by d(Ir) and pi(carbene). The LUMOs (lowest unoccupied molecular orbitals) of 1, 2, 4, and 5 are localized on carbene, but that of 3 is localized on acac. The calculated lowest-lying absorptions with TD-DFT method based on Perdew-Burke-Erzenrhof (PBE) functional of 1 (310 nm), 2 (332 nm), and 3 (347 nm) have ML(carbene)CT/IL(phenyl ->carbene)CT (MLCT = metal-to-ligand charge transfer; ILCT = intraligand charge transfer) transition characters, whereas those of 4 (385 nm) and 5 (389 nm) are assigned to ML(carbene)CT/IL(carbene-->carbene)CT transitions. The phosphorescences calculated by TD-DFT method with PBE0 functional of 1 (386 nm) and 2 (388 nm) originate from (3)ML(carbene)CT/(3)IL(phenyl-->carbene)CT excited states, but those of 4 (575 nm) and 5 (578 nm) come from (3)ML(carbene)CT/(3)IL(carbene-->carbene)CT excited states. The calculated results showed that the carbene and phenyl groups act as two independent chromophores in transition processes. Compared with 1 and 2, the absorptions of 4 and 5 are red-shifted by increasing the effective pi-conjugation groups near the C(carbene) atom. We predicated that (pmb)(2)Ir(acac) is nonemissive, because the LUMO of 3 is contributed by the nonemissive acac ligand. PMID- 19551886 TI - Chloride ion conduction without water coordination in the pore of ClC protein. AB - In the present work, we have found by an atomistic molecular dynamics simulation that hydrogen atoms originating from the residues of a prokaryotic ClC protein (EcClC) stabilize the chloride ion without water molecules in the pore of ClC protein. When the chloride ion conduction is simulated by pulling a chloride ion along the pore axis, the free energy barrier for chloride ion conduction is calculated to be low (4 kcal/mol), although the chloride ion is stripped of its hydration shell as it passes through the dehydrated pore region. The calculation of the number of hydrogen atoms surrounding the chloride ion reveals that water molecules hydrating the chloride ion are replaced by polar and non-polar hydrogen atoms protruding from the protein residues. From the analysis of the pair interaction energy between the chloride ion and these hydrogen atoms, it is realized that the hydrogen atoms from the protein residues stabilize the chloride ion at the dehydrated region instead of water molecules, by which the energetic penalty for detaching water molecules from the permeating ion is compensated. PMID- 19551888 TI - Sulfite pretreatment to overcome recalcitrance of lignocellulose (SPORL) for robust enzymatic saccharification of hardwoods. AB - This study demonstrates sulfite pretreatment to overcome recalcitrance of lignocellulose (SPORL) for robust bioconversion of hardwoods. With only about 4% sodium bisulfite charge on aspen and 30-min pretreatment at temperature 180 degrees C, SPORL can achieve near-complete cellulose conversion to glucose in a wide range of pretreatment liquor of pH 2.0-4.5 in only about 10 h enzymatic hydrolysis. The enzyme loading was about 20 FPU cellulase plus 30 CBU beta glucosidase per gram of cellulose. The production of fermentation inhibitor furfural was less than 20 mg/g of aspen wood at pH 4.5. With pH 4.5, SPORL avoided reactor corrosion problem and eliminated the need for substrate neutralization prior to enzymatic hydrolysis. Similar results were obtained from maple and eucalyptus. PMID- 19551887 TI - Mental status changes after hematopoietic stem cell transplantation. AB - BACKGROUND: The growing numbers of survivors of innovative cancer treatments, such as hematopoietic stem cell transplantation (HSCT), often report subsequent cognitive difficulties. The objective of this study was to evaluate and compare neurocognitive changes in patients with chronic myelogenous leukemia (CML) or primary myelodysplastic syndrome (MDS) after allogeneic HSCT or other therapies. METHODS: In this prospective cohort study, serial evaluations of attention, concentration, memory, mood, and quality of life were used in a consecutive sample of 106 eligible patients who had CML (n = 91) or MDS (n = 15) at enrollment and then 12 months and 18 months after HSCT or other therapy. RESULTS: The 3 evaluations at enrollment, 12 months, and 18 months were completed by 98%, 95%, and 89% of surviving participants, respectively. Among all patients, there was significant improvement in memory over 18 months. For example, the 45 patients who underwent HSCT (42 patients with CML and 3 patients with MDS) compared favorably with the patients who received other treatment on most measures of neuropsychological function, except they had improved mental health (P = .034), worse physical function (P = .049), and more difficulty with coordination and fine motor speed bilaterally (dominant hand, P = .005; nondominant hand, P = .0019). Patients with CML overall had improved phonemic fluency (P = .014). CONCLUSIONS: The current study indicated that time and diagnosis may be important factors when assessing neurocognitive and other changes. Complaints regarding "chemobrain" after HSCT merit further study, because deficits actually may predate the initiation of treatment and subsequently may improve. The study results could reassure prospective HSCT recipients, because HSCT compared favorably with other treatments when mental status side effects were considered. PMID- 19551890 TI - Rapid algal toxicity assay using variable chlorophyll fluorescence for Chlorella kessleri (chlorophyta). AB - Three methods of algal assays--the standard assay, microassay, and the proposed fluorescence assay--are compared from the point of view of reliability of EC50 detection, the minimum required time for the detection, sensitivity of individual measurement, i.e. at which cell density the particular assay can be used for EC50 estimation, and the time stability of the EC50 values. The assays were performed with green alga Chlorella kessleri strain LARG/1 growing in potassium dichromate solution in Z-medium ranging from 0.01 to 100 mg Cr L-1. The inoculation cell density was set according to the standards to 104 cells mL-1 and according to spectrophotometer/plate reader detection limit. The average EC50 ranged from 0.096 to 0.649 mg Cr L-1 and there were no significant differences in EC50 between the assay type and the inoculation methods with the exception of the significant difference between EC(c)5072 (EC50 established from biomass measured as chlorophyll a concentration after 72 h of cultivation) in the standard assay and EC(r)50 (EC50 derived from growth rate) in the microassay in the standard inoculation experiment due to low variability of their values. The EC(f)50 (EC50 derived from variable fluorescence measurement) values correspond to EC50 values derived from the growth rates. Fluorescence measurement revealed the toxic effect of the chromium after 24 h of exposure at cell density of 5 x 104 cells mL-1, less by half than other used assay methods. The positive correlation of EC(f)50 and time was found in the standard inoculation experiment but opposite effect was observed at the spectrophotometric one. PMID- 19551891 TI - Invasion characteristics of oral tongue cancer: frequency of reporting and effect on survival in a population-based study. AB - BACKGROUND: The 2000 College of American Pathologists (CAP) guidelines recommend that a characterization of carcinomas of the upper aerodigestive tract, including tongue cancer, should include depth of invasion (DI) and the presence of lymphovascular invasion (LVI) or perineural invasion (PNI). METHODS: This study included patients who were diagnosed with cancer of the oral tongue, who underwent tumor resection, and who were reported to either the Metropolitan Atlanta and Rural Georgia Surveillance, Epidemiology, and End Results (SEER) registry or the Los Angeles SEER registry. The authors assessed the completeness of pathology reporting with respect to the documentation of PNI or LVI and DI. Generalized estimating equations were used to examine factors that influenced reporting while taking into consideration clustering of observations within the hospitals. Univariate and multivariate survival analyses were conducted to examine the impact of tumor invasion characteristics on mortality while controlling for other prognostic factors. RESULTS: DI reporting increased from 13% between 1997 and 1999 to 23% between 2000 and 2004 after the CAP issued its recommendations; whereas mode of invasion (the presence of LVI and/or PNI) reporting for the same period increased from 13% to 38%. The observed increase in reporting was most pronounced in the first 2 years (2000 and 2001) and appeared to decline again afterward. Tumor invasion >3 mm in depth and the presence of PNI were among the strongest predictors of survival in multivariate analyses. CONCLUSIONS: The current results indicated the importance of reporting tumor invasion characteristics for patients diagnosed with cancer of the oral tongue. The findings also underscore the need for continuous monitoring of adherence to the CAP protocol. PMID- 19551889 TI - Yes-associated protein is an independent prognostic marker in hepatocellular carcinoma. AB - BACKGROUND: Yes-associated protein (YAP), a downstream target of the Hippo signaling pathway, was recently linked to hepatocarcinogenesis in a mouse hepatocellular carcinoma (HCC) model. The objective of the current study was to investigate the clinical significance of YAP in HCC and its prognostic values in predicting survival and tumor recurrence. METHODS: The authors collected 177 pairs of tumor and adjacent nontumor tissue from HCC patients with definitive clinicopathologic and follow-up data. YAP expression was determined by immunohistochemistry, Western blot analysis, and quantitative polymerase chain reaction. Association of YAP with each clinicopathologic feature was analyzed by Pearson chi-square test, and HCC-specific disease-free survival and overall survival by Kaplan-Meier curves and log-rank test. Multivariate Cox regression analyses of YAP in HCC were also performed. RESULTS: YAP was expressed in the majority of HCC cases (approximately 62%) and mainly accumulated in the tumor nucleus. Overexpression of YAP in HCC was significantly associated with poorer tumor differentiation (Edmonson grade; P = .021) and high serum alpha-fetoprotein (AFP) level (P < .001). Kaplan-Meier and Cox regression data indicated that YAP was an independent predictor for HCC-specific disease-free survival (hazards ratio [HR], 1.653; 95% confidence interval [95% CI], 1.081-2.528 [P = .02]) and overall survival (HR, 2.148; 95% CI, 1.255-3.677 [P = .005]). CONCLUSIONS: YAP is an independent prognostic marker for overall survival and disease-free survival times of HCC patients and clinicopathologically associated with tumor differentiation and serum AFP level. It is a potential therapeutic target for this aggressive malignancy. PMID- 19551892 TI - p53-induced RING-H2 protein, a novel marker for poor survival in hepatocellular carcinoma after hepatic resection. AB - BACKGROUND: Currently, the role of p53-induced RING-H2 protein (PIRH2) in the development of hepatocellular carcinoma (HCC) remains unknown. The objective of this retrospective study was to investigate the expression of PIRH2 and its relation to prognosis in patients with HCC after hepatic resection. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR), quantitative real-time RT-PCR, and Western blot analyses were used to detect expression levels of PIRH2 in 30 samples of HCC tissue and paracarcinomatous liver tissue (PCLT) and in 5 samples of normal liver tissue (NL). In addition, immunohistochemical analysis was performed on 122 HCC specimens and follow-up information data from those patients were reviewed. RESULTS: Both messenger RNA and protein expression levels of PIRH2 were elevated significantly in HCC tissues compared with PCLT and NL tissues. The increased PIRH2 expression was correlated with vein invasion, Edmondson-Steiner grade, TNM stage, and multiple tumor nodes (P<.05). It is noteworthy that the patients with HCC who had high PIRH2 expression had shorter overall survival and disease-free survival than the patients who had low PIRH2 expression (median survival, 280 days vs 372 days; P = .0002; median disease-free survival, 220 days vs 310 days; P = .0016). Multivariate Cox regression analysis revealed that high PIRH2 expression was an independent prognostic factor for patients with HCC (relative risk, 1.792; P = .009). CONCLUSIONS: The current data revealed that increased expression of PIRH2 was correlated with poor survival in patients with HCC, indicating that PIRH2 is a novel prognostic marker for HCC. PMID- 19551893 TI - Real-time visualization of oxidative stress in a floating macrophyte Lemna minor L. exposed to cadmium, copper, menadione, and AAPH. AB - An ultra-sensitive digital imaging system was employed to visualize oxidative stress in intact L. minor plants exposed to Cd, Cu, menadione, AAPH, and ascorbate in real time. The increase of ROS production was assessed by measuring the rate of fluorescence intensity increases of the test medium supplemented with a fluorescing probe (dichlorofluorescein diacetate). The addition of 100 MUM CdCl2 or 100 MUM CuSO4 to the growth medium resulted in a significant increase of medium fluorescence. Additionally, CuSO4 caused a significantly higher fluorescence intensity than CdCl2 did. A strong positive correlation (R2 = 0.99) between menadione concentration and fluorescence intensity was observed. The positive correlation between AAPH concentration and fluorescence intensity was not as strong as in the case of menadione (R2 = 0.81). Menadione induced a stronger oxidative stress than similar concentration of AAPH. The addition of 100 MUM ascorbate to L. minor treated with 50 MUM menadione significantly reduced the fluorescence intensity increase. A linear trend of the fluorescence increase was observed in all treatments, indicating that chemical-induced oxidative stress is a gradual process and that the applied concentrations of the chemicals caused a constant increased production of ROS with different intensities, depending on the treatment. This is the combined result of a gradual diminishing of antioxidant reserves and accumulating oxidative damage. The observed rates of ROS production were slower than those in the studies using cell cultures. PMID- 19551894 TI - Glycosylation of aromatic amines III: Mechanistic implications of the pH dependent glycosylation of various aromatic amines (kynurenine, 2' aminoacetophenone, daptomycin, and sulfamethoxzaole). AB - Glycosylation reaction kinetics of a series of aromatic amines (kynurenine, 2' aminoacetophenone, daptomycin, and sulfamethoxazole) was compared to propose a unifying reaction mechanism. Kinetic studies were conducted in aqueous solutions containing glucose in the pH range 1-6.5 with 2'-aminoacetophenone and daptomycin. The resultant pH-rate profiles were compared to previously reported profiles for the reactions of glucose and kynurenine or sulfamethoxazole. Glycosylation of weakly basic aromatic amines involved the addition of the unprotonated amine to the aldehydic sugar leading to carbinolamine formation followed by specific acid catalyzed dehydration. All of the pH-rate profiles displayed characteristic downward bend at pH 4-5 due to a change from rate determining addition to dehydration. In the pH-rate profile for kynurenine, a second downward bend was observed in the pH region 2-4. This feature was absent for the other substrates and was attributed to differences in reactivity of the two ionization states of the alpha carboxylic acid in kynurenine. This stabilization was not possible for the other amines studied. PMID- 19551895 TI - Cytomics and regenerative medicine. PMID- 19551896 TI - Water polygons in high-resolution protein crystal structures. AB - We have analyzed the interstitial water (ISW) structures in 1500 protein crystal structures deposited in the Protein Data Bank that have greater than 1.5 A resolution with less than 90% sequence similarity with each other. We observed varieties of polygonal water structures composed of three to eight water molecules. These polygons may represent the time- and space-averaged structures of "stable" water oligomers present in liquid water, and their presence as well as relative population may be relevant in understanding physical properties of liquid water at a given temperature. On an average, 13% of ISWs are localized enough to be visible by X-ray diffraction. Of those, averages of 78% are water molecules in the first water layer on the protein surface. Of the localized ISWs beyond the first layer, almost half of them form water polygons such as trigons, tetragons, as well as expected pentagons, hexagons, higher polygons, partial dodecahedrons, and disordered networks. Most of the octagons and nanogons are formed by fusion of smaller polygons. The trigons are most commonly observed. We suggest that our observation provides an experimental basis for including these water polygon structures in correlating and predicting various water properties in liquid state. PMID- 19551898 TI - WITHDRAWN: impaired spermatogenesis in mice overexpressing stem cell protein Piwil2 (Mili). PMID- 19551897 TI - Regulation of prostaglandin synthesis in ovaries of sexually-mature zebrafish (Danio rerio). AB - This study investigates the regulation of prostaglandin (PG) synthesis in the ovaries of sexually-mature zebrafish (Danio rerio). We examined the ovarian expression of genes within the arachidonic acid (AA) pathway, and the ovarian levels of 17alpha,20beta-dihydroxy-4-pregnen-3-one (17alpha,20beta-P), 17beta estradiol (E(2)), and PGF(2alpha) in spawning and nonspawning fish during the ovulatory cycle. Real-time RT-PCR analysis revealed that the expression levels of cytosolic phospholipase A(2) (cpla2) and cyclooxygenases (COX)-2 (ptgs2) in ovarian fragments and in isolated full-grown follicles of spawning fish were highest at 6:00 when ovulation was expected to occur. In nonspawning fish, cpla2 expression levels declined over time while ptgs2 expression displayed the same temporal pattern as in spawning fish. Elevated levels of 17alpha,20beta-P in the spawning fish occurred at 3:30, but there were no changes in the nonspawning fish. In other studies conducted to investigate the hormonal regulation of AA pathway genes, fish exposed via the water for 24 or 96 hr to 17alpha,20beta-P or E(2) exhibited reduced ovarian expression levels of COX-1 (ptgs1) and PG E synthase-2 (ptgsl), and E(2) reduced the expression of cpla2. Injection of human chorionic gonadotropin (hCG) (100 IU) led to increased expression levels of cpla2 and ptgs2 at 2 and 18 hr post-treatment, but consistently reduced ptgs1 and ptgsl expression. In these fish, ovarian levels of 17alpha,20beta-P were elevated at all time points and PGF(2alpha) levels in the hCG-treated group were significantly higher than the control fish at 18 hr. Collectively, these in vivo results suggest that gonadotropins and steroids are involved in the regulation of the AA pathway in ovarian follicles of zebrafish. PMID- 19551900 TI - The TLR-independent DNA recognition pathway in murine macrophages: Ligand features and molecular signature. AB - Recognition of foreign DNA by cytosolic innate immune receptors triggers the production of IFN-beta. However, it is unclear whether different types of DNA ligands are recognized by similar receptors and whether the resulting response is distinct from the endosomal TLR response. To address these questions, we compared the two most commonly used types of DNA ligands (IFN-stimulatory DNA (ISD) and poly(dAdT)) and assessed the minimal structural requirements for stimulatory capacity in RAW264.7 cells. Gene expression signatures and competition experiments suggest that ISD and poly(dAdT) are qualitatively indistinguishable and differ from the CpG-containing oligonucleotides triggering the TLR9 pathway. Structure - activity relationship analyses revealed that a minimal length of two helical turns is sufficient for ISD-mediated IFN-beta induction, while phosphorylation at the 5'-end is dispensable. Altogether, our data suggest that, in murine macrophages, only one major cytosolic DNA recognition pathway is operational. PMID- 19551899 TI - Lack of TNFR2 expression by CD4(+) T cells exacerbates experimental colitis. AB - TNF plays fundamental roles in the induction and perpetuation of inflammation. The effects of TNF are mediated through TNF receptor (TNFR) 1 or 2. As these two receptors mediate different functions, selective targeting of one receptor may represent a more specific treatment for inflammatory disorders than the complete blocking of TNF. TNFR2 expression is up-regulated in inflammatory bowel disease. Hence, we directly assessed the role of TNFR2 signaling in the CD4(+) T-cell transfer model of colitis using TNFR2(-/-) or WT mice as donors of colitogenic CD4(+)CD45RB(hi) T cells for transfer into syngeneic RAG2(-/-) or RAG2(-/-)TNFR2( /-) recipient mice. Although the absence of TNFR2 expression by non-lymphoid cells of the recipient mice does not influence the course of colitis, transfer of TNFR2(-/-) CD4(+) T cells leads to an accelerated onset of disease and to more severe signs of inflammation. The enhanced colitogenic potential of TNFR2(-/-) CD4(+) T cells is associated with reduced activation-induced cell death, resulting in an increased accumulation of TNFR2(-/-) CD4(+) T cells. Hence, TNFR2 signaling is crucial for the TNF-dependent contraction of the disease-inducing T cells. Therefore, a selective blocking of TNFR2 may lead to exacerbation rather than attenuation of T-cell-mediated inflammatory disorders. PMID- 19551901 TI - Progress technology in microencapsulation methods for cell therapy. AB - Cell encapsulation in microcapsules allows the in situ delivery of secreted proteins to treat different pathological conditions. Spherical microcapsules offer optimal surface-to-volume ratio for protein and nutrient diffusion, and thus, cell viability. This technology permits cell survival along with protein secretion activity upon appropriate host stimuli without the deleterious effects of immunosuppressant drugs. Microcapsules can be classified in 3 categories: matrix-core/shell microcapsules, liquid-core/shell microcapsules, and cells core/shell microcapsules (or conformal coating). Many preparation techniques using natural or synthetic polymers as well as inorganic compounds have been reported. Matrix-core/shell microcapsules in which cells are hydrogel-embedded, exemplified by alginates capsule, is by far the most studied method. Numerous refinement of the technique have been proposed over the years such as better material characterization and purification, improvements in microbead generation methods, and new microbeads coating techniques. Other approaches, based on liquid core capsules showed improved protein production and increased cell survival. But aside those more traditional techniques, new techniques are emerging in response to shortcomings of existing methods. More recently, direct cell aggregate coating have been proposed to minimize membrane thickness and implants size. Microcapsule performances are largely dictated by the physicochemical properties of the materials and the preparation techniques employed. Despite numerous promising pre clinical results, at the present time each methods proposed need further improvements before reaching the clinical phase. PMID- 19551902 TI - Pathway sensitivity analysis for detecting pro-proliferation activities of oncogenes and tumor suppressors of epidermal growth factor receptor-extracellular signal-regulated protein kinase pathway at altered protein levels. AB - BACKGROUND: Mathematic models and sensitivity analyses of biologic pathways have been used for exploring the dynamics and for detecting the key components of signaling pathways. METHODS: The authors previously developed a mathematic model of the epidermal growth factor receptor-extracellular signal-regulated protein kinase (EGFR-ERK) pathway using ordinary differential equations from existing EGFR-ERK pathway models. By using prolonged ERK activation as an indicator that may lead to cell proliferation under certain circumstances, in the current study, a pathway sensitivity analysis was performed to test its capability of detecting pro-proliferative activities through altered protein levels to examine the effects on ERK activation. RESULTS: The analysis revealed that 12 of 20 oncoproteins and 4 of 5 tumor suppressors were detected, consistent with reported experimental works. Because pathway dynamics depend on many factors, some of which were not included in the current models, failure to detect all known oncogenes and tumor suppressors can be because of the failure to include relevant crosstalk to other pathway components. CONCLUSIONS: Overall, the current results indicated that pathway sensitivity analysis is a useful approach for detecting and distinguishing pro-proliferation activities of oncoproteins and suppressed proliferative activities of tumor suppressors at altered protein levels at least in the EGFR-ERK model. PMID- 19551905 TI - Validation of beta-actin used as endogenous control for gene expression analysis in mechanobiology studies. PMID- 19551906 TI - A splice site mutation combined with a novel missense mutation of LHCGR cause male pseudohermaphroditism. AB - Leydig cell hypoplasia (LCH) is a rare form of male pseudohermaphroditism caused by inactivating mutations in the luteinizing hormone receptor gene (LHCGR). The majority of LHCGR mutations are located in the coding sequence, resulting in impairment of either LH/CG binding or signal transduction. We report a Chinese family with two siblings (46, XY and 46, XX) carrying a missense mutation (c. 455 T>C, p. Ile152Thr) and a splice site mutation (c. 537-3 C>A). Computational analysis of the missense mutation in the three-dimensional structural model predicted it might influence the distribution of hydrogen bonds and intermolecular contacts between the hormone and receptor. Consistent with these findings, in vitro mutant analysis revealed a marked impairment of human chorionic gonadotropin binding and signal transduction. The splice-acceptor mutation (c. 537-3 C>A) resulted in abnormal splicing of LHCGR mRNA, skipping exon 7. This report expands the genotypic spectrum of LHCGR mutations, with relevant implications for the molecular analysis of this gene. PMID- 19551907 TI - Notch2 signaling promotes biliary epithelial cell fate specification and tubulogenesis during bile duct development in mice. AB - Intrahepatic bile duct (IHBD) development begins with the differentiation of hepatoblasts into a single continuous biliary epithelial cell (BEC) layer, called the ductal plate. During ductal plate remodeling, tubular structures arise at distinct sites of the ductal plate, forming bile ducts that dilate into the biliary tree. Alagille syndrome patients, who suffer from bile duct paucity, carry Jagged1 and Notch2 mutations, indicating that Notch2 signaling is important for IHBD development. To clarify the role of Notch2 in BEC differentiation, tubulogenesis, and BEC survival, we developed a mouse model for conditional expression of activated Notch2 in the liver. We show that expression of the intracellular domain of Notch2 (Notch2ICD) differentiates hepatoblasts into BECs, which form additional bile ducts in periportal regions and ectopic ducts in lobular regions. Additional ducts in periportal regions are maintained into adulthood and connect to the biliary tight junction network, resulting in an increased number of bile ducts per portal tract. Remarkably, Notch2ICD-expressing ductal plate remnants were not eliminated during postnatal development, implicating Notch2 signaling in BEC survival. Ectopic ducts in lobular regions did not persist into adulthood, indicating that local signals in the portal environment are important for maintaining bile ducts. CONCLUSION: Notch2 signaling regulates BEC differentiation, the induction of tubulogenesis during IHBD development, and BEC survival. PMID- 19551909 TI - AM-FM techniques in the analysis of optical coherence tomography signals. AB - The subtle tissue changes associated with the early stages of malignancies, such as cancer, are not clearly discernible even at the current, improved, resolution of optical coherence tomography (OCT) systems. However, these changes directly affect the spectral content of the OCT image that contains information regarding these unresolvable features. Spectral analysis of OCT signals has recently been shown to provide additional information, resulting in improved contrast, directly related to scatterer size changes. Amplitude modulation-frequency modulation (AM FM) analysis, a fast and accurate technique for the estimation of the instantaneous frequency, phase, and amplitude of a signal, can also be applied to OCT images to extract scatterer-size information. The proposed technique could make available an extremely valuable tool for the investigation of disease characteristics that now remain below the resolution of OCT and could significantly improve the technology's diagnostic capabilities. PMID- 19551904 TI - Genetic basis of inherited macular dystrophies and implications for stem cell therapy. AB - Untreatable hereditary macular dystrophy (HMD) presents a major burden to society in terms of the resulting patient disability and the cost to the healthcare provision system. HMD results in central vision loss in humans sufficiently severe for blind registration, and key issues in the development of therapeutic strategies to target these conditions are greater understanding of the causes of photoreceptor loss and the development of restorative procedures. More effective and precise analytical techniques coupled to the development of transgenic models of disease have led to a prolific growth in the identification and our understanding of the genetic mutations that underly HMD. Recent successes in driving differentiation of pluripotent cells towards specific somatic lineages have led to the development of more efficient protocols that can yield enriched populations of a desired phenotype. Retinal pigmented epithelial cells and photoreceptors derived from these are some of the most promising cells that may soon be used in the treatment of specific HMD, especially since rapid developments in the field of induced pluripotency have now set the stage for the production of patient-derived stem cells that overcome the ethical and methodological issues surrounding the use of embryonic derivatives. In this review we highlight a selection of HMD which appear suitable candidates for combinatorial restorative therapy, focusing specifically on where those photoreceptor loss occurs. This technology, along with increased genetic screening, opens up an entirely new pathway to restore vision in patients affected by HMD. PMID- 19551908 TI - Human immunodeficiency virus and hepatitis C infections induce distinct immunologic imprints in peripheral mononuclear cells. AB - Coinfection with hepatitis C virus (HCV) is present in one-third of all human immunodeficiency virus (HIV)-infected individuals in the United States and is associated with rapid progression of liver fibrosis and poor response to pegylated interferon (IFN) and ribavirin. In this study we examined gene expression profiles in peripheral blood mononuclear cells (PBMCs) from different groups of individuals who are monoinfected or coinfected with HIV and HCV. Data showed that HIV and HCV viremia up-regulate genes associated with immune activation and immunoregulatory pathways. HCV viremia is also associated with abnormalities in all peripheral immune cells, suggesting a global effect of HCV on the immune system. Interferon-alpha-induced genes were expressed at a higher level in PBMCs from HIV-infected individuals. HCV and HIV infections leave distinct profiles or gene expression of immune activation in PBMCs. HIV viremia induces an immune activated state; by comparison, HCV infection induces immunoregulatory and proinflammatory pathways that may contribute to progression of liver fibrosis. CONCLUSION: An aberrant type-I IFN response seen exclusively in HIV-infected individuals could be responsible for the poor therapeutic response experienced by HIV/HCV coinfected individuals receiving interferon-alpha based current standard of care. PMID- 19551911 TI - Influence of the solvent ability to form hydrogen bonds in the crystal structure of ([3beta,5beta,7alpha,12alpha]-3[(norbornyl-2-acetyl)-amino]-7,12 dihydroxycholan-24-oic acid (a norbornyl derivative of cholic acid). AB - A norbornyl-2-acetyl derivative of cholic acid ([3beta,5beta,7alpha,12alpha] 3[(norbornyl-2-acetyl)-amino]-7,12-dihydroxycholan-24-oic acid -NbCH2CA-) was synthesized and recrystallized in two dipolar aprotic solvents (acetone, DMSO) and in one protic solvent (2-propanol). In DMSO and acetone the crystals are orthorhombic, P2(1)2(1)2(1) (all their parameters being very similar) while in 2 propanol the crystal is monoclinic, P2(1). The inclusion complexes with the solvent have a 1:1 stochiometry with DMSO and acetone and 1:2 with 2-propanol. All solvents are forming a hydrogen bond with the amide bond of the bridge between the norbornyl residue and the steroid nucleus of the bile acid. In DMSO and acetone the beta side of the steroid groups lies in the same region facilitating hydrophobic interactions, and the molecules are disposed in an antiparallel orientation (the methyl groups having a beta interdigitation) forming bilayers. The width of the bilayers is 9.231 A and 8.859 A in DMSO and acetone, respectively. A lamellar structure is also evident for the crystal in 2 propanol (the width being 11.908 A), but the packing is different from the previous one since a sliding between the steroid groups is observed and the methyl groups are not interdigitated. Four different hydrogen bonds are established by every steroid molecule in the NbCH2CA/DMSO (or acetone) crystal. This hydrogen bond network interconnects the hydrophilic regions of the lamellar structure. The hydrogen bond network of the NbCH2CA:2-propanol crystal is different because of the different abilities of 2-propanol to form hydrogen bonds. The side chain has a ttti conformation in the two orthorhombic crystals, and a tgtg one in the monoclinic crystal. PMID- 19551912 TI - Proceedings of the Xth International Plant Virus Epidemiology Symposium. October 14-18, 2007. Patancheru, Hyderabad, Andhra Pradesh, India. PMID- 19551913 TI - Confusion over counterfeit drugs in Uganda. PMID- 19551914 TI - Raw deal for refused asylum seekers in the UK. PMID- 19551915 TI - Tuberculosis: pictures of a ticking time bomb. PMID- 19551916 TI - Valuing communication. PMID- 19551917 TI - FGLamide Allatostatin genes in Arthropoda: introns early or late? AB - FGLamide allatostatins are invertebrate neuropeptides which inhibit juvenile hormone biosynthesis in Dictyoptera and related orders and also show myomodulatory activity. The FGLamide allatostatin (AST) gene structure in Dictyoptera is intronless within the ORF, whereas in 9 species of Diptera, the FGLamide AST ORF has one intron. To investigate the evolutionary history of AST intron structure, (intron early versus intron late hypothesis), all available Arthropoda FGLamide AST gene sequences were examined from genome databases with reference to intron presence and position/phase. Three types of FGLamide AST ORF organization were found: intronless in I. scapularis and P. humanus corporis; one intron in D. pulex, A. pisum, A. mellifera and five Drosophila sp.; two introns in N. vitripennis, B. mori strains, A. aegypti, A. gambiae and C. quinquefasciatus. The literature suggests that for the majority of genes examined, most introns exist between codons (phase 0) which may reflect an ancient function of introns to separate protein modules. 60% of the FGLamide AST ORFs introns were between the first and second base within a codon (phase 1), 28% were between the second and third nucleotides within a codon (phase two) and 12% were phase 0. As would be required for correct intron splicing consensus sequence, 84% of introns were in codons starting with guanine. The positioning of introns was a maximum of 9 codons from a dibasic cleavage site. Our results suggest that the introns in the analyzed species support the intron late model. PMID- 19551918 TI - Recurrence of the 'deep-intronic' FGG IVS6-320A>T mutation causing quantitative fibrinogen deficiency in the Italian population of Veneto. AB - Quantitative fibrinogen deficiency is a rare bleeding disorder characterized by abnormally low levels of fibrinogen in plasma, generally due to mutations in one of the three fibrinogen genes: FGA, FGB, and FGG, coding for A alpha, B beta, and gamma chain, respectively. Although the partial defect (hypofibrinogenemia) is due to mutations occurring in the heterozygous state, homozygosity or compound heterozygosity for the same genetic defects give rise to the more severe afibrinogenemia. Mutations responsible for these conditions are scattered throughout the three fibrinogen genes, with only few sites representing relative mutational hot spots. In this study, we report the identification of the FGG IVS6 320A>T mutation in an Italian hypofibrinogenemic patient from Veneto (a region of North-Eastern Italy). This 'deep-intronic' mutation, which would go unnoticed by using conventional mutational screening strategies was previously reported in an afibrinogenemic family from Vicenza (a province of Veneto). The geographic clustering of patients carrying the FGG IVS6-320A>T mutation and the results of haplotype analysis suggest the existence of a common founder. This information will be useful to direct future genetic screenings in patients coming from the same geographic area. PMID- 19551919 TI - Teens and drug facilitated sexual assault. PMID- 19551920 TI - Prognostic factors in gastrointestinal bleeding due to peptic ulcer, construction of a predictive model. PMID- 19551922 TI - 30-year-old HIV-positive female with diffuse alveolar hemorrhage. AB - We present a case of Strongyloides stercoralis hyperinfection in a 30-year-old HIV positive female who presents with diffuse alveolar hemorrhage. We discuss the relevant differential diagnoses and characteristic imaging findings. PMID- 19551921 TI - Design and development of a cardiopulmonary resuscitation mattress. AB - This study introduces the design and construction of a mattress insert to produce more effective cardiopulmonary resuscitation (CPR). The mattress insert deflates, making the mattress insert a rigid surface. Using a device that administers a constant compression depth onto a manikin, we were able to show that our mattress insert more effectively directed the compressive force to the manikin compared to the current practice of using a headboard on top of a mattress. The mattress insert produced a statistically significant increase in the compression efficiency when compared to the current practice of using the headboard (81% vs. 53%). Because the mattress insert starts deflating immediately after the vacuum is turned on, 1 person is needed to initiate chest compressions. Compression begins while the mattress deflates. Running compression tests at incremental time periods, we found that our design reaches and surpasses the compression efficiency when using a headboard in 10 seconds. PMID- 19551923 TI - Policy, politics, & nursing practice. In this issue. PMID- 19551924 TI - The clothes make the iceman. PMID- 19551925 TI - Model independent determination of colloidal silica size distributions via analytical ultracentrifugation. AB - We report a method to determine the particle size distribution of small colloidal silica spheres via analytical ultracentrifugation and show that the average particle size, variance, standard deviation, and relative polydispersity can be obtained from a single sedimentation velocity (SV) analytical ultracentrifugation (AUC) experiment. The particle size distribution (psd) from the enhanced van Holde-Weischet (vHW) analysis accounts for the dynamic light scattering results quite well. In addition, the vHW psd equals the psd from a continuous distribution of sedimentation coefficients analysis where whole sedimentation velocity boundaries are fitted. The SV AUC interference optical data also yield the specific particle volume such that distributions of sedimentation coefficients for colloidal spheres can be converted directly to particle size distributions. Our results show that SV AUC experiments may yield a quantitative particle size distribution without a priori knowledge of the particle size and the shape of the size distribution. PMID- 19551926 TI - Natural organic matter and the event horizon of mass spectrometry. AB - Soils, sediments, freshwaters, and marine waters contain natural organic matter (NOM), an exceedingly complex mixture of organic compounds that collectively exhibit a nearly continuous range of properties (size-reactivity continuum). NOM is composed mainly of carbon, hydrogen, and oxygen, with minor contributions from heteroatoms such as nitrogen, sulfur, and phosphorus. Suwannee River fulvic acid (SuwFA) is a fraction of NOM that is relatively depleted in heteroatoms. Ultrahigh resolution Fourier transform ion cyclotron (FTICR) mass spectra of SuwFA reveal several thousand molecular formulas, corresponding in turn to several hundred thousand distinct chemical environments of carbon even without accountancy of isomers. The mass difference deltam among adjoining C,H,O molecules between and within clusters of nominal mass is inversely related to molecular dissimilarity: any decrease of deltam imposes an ever growing mandatory difference in molecular composition. Molecular formulas that are expected for likely biochemical precursor molecules are notably absent from these spectra, indicating that SuwFA is the product of diagenetic reactions that have altered the major components of biomass beyond the point of recognition. The degree of complexity of SuwFA can be brought into sharp focus through comparison with the theoretical limits of chemical complexity, as constrained and quantized by the fundamentals of chemical binding. The theoretical C,H,O-compositional space denotes the isomer-filtered complement of the entire, very vast space of molecular structures composed solely of carbon, hydrogen, and oxygen. The molecular formulas within SuwFA occupy a sizable proportion of the theoretical C,H,O-compositional space. A 100 percent coverage of the theoretically feasible C,H,O-compositional space by SuwFA molecules is attained throughout a sizable range of mass and H/C and O/C elemental ratios. The substantial differences between (and complementarity of) the SuwFA molecular formulas that are observed using six different modes of ionization (APCI, APPI, and ESI in positive and negative modus) imply considerable selectivity of the ionization process and suggest that the observed mass spectra represent simplified projections of still more complex mixtures. PMID- 19551927 TI - Trapping of bioparticles via microvortices in a microfluidic device for bioassay applications. AB - This paper presents hydrodynamic trapping of bioparticles in a microfluidic device. An in-plane oscillatory microplate, driven via Lorentz law, generates two counter-rotating microvortices, trapping the bioparticles within the confines of the microvortices. The force required to trap bioparticles is quantified by tuning the background flow and the microplate's excitation voltage. Trapping and releasing of 10-microm polystyrene beads, human embryonic kidney (HEK) cells, red blood cells (RBCs), and IgG antibodies were demonstrated. Results show the microvortices rotates at 0-6 Hz corresponding to 2-9 Vpp (peak-to-peak) excitation. At a particular rate of rotation (2-7 Vpp tested), a bioparticle is trapped until the background flow exceeds a limit. This flow limit increases with the rate of rotation, which defines the trap/release force boundary over the range of operation. This boundary is 12 +/- 2.0 pN for cell-size bioparticles and 160 +/- 50 fN for antibodies. Trapping of RBCs demonstrated microvortices' ability for nonspherical cells. Cell viability was studied via HEK cells that were trapped for 30 min and shown to be viable. This hydrodynamically controlled approach to trap a wide range of bioparticles should be useful as a microfluidic device for cellular and subcellular bioassay applications. PMID- 19551928 TI - Evaluating low-intensity unknown signals in quantitative proton NMR mixture analysis. AB - Analytical analyses of highly complex mixtures, such as biofluids or liquid food products, often give rise to signals for unknown compounds, particularly for compounds at low concentration. Here we compare two conventional chemometric approaches for NMR spectral analysis ("spectral binning" and "high-resolution analysis") with a novel library-based method ("targeted profiling of unknowns", TPU). The three methods were applied to a proton NMR spectral data set of ultrafiltered mouse serum typical of those examined in metabolomics/metabonomics studies. The advantages of high-resolution analysis of typical NMR peaks have been well described previously, and as a result we examined low intensity unknowns peaks (LIUPs). A total of 25 LIUPs were assessed based on their significance to multivariate statistical analysis of the data set using the TPU method. The linearity of NMR signals at low incremental concentration changes (< 10 microM) was determined by titration of endogenously occurring metabolites into filtered mouse serum. Carbon-13 decoupling of the NMR spectra was used to ensure isotope-satellite peaks were eliminated. Four peaks were noted as significant to separation between arthritic and diseased animals. The conventional spectral methods were hampered by baseline noise or overlap with high concentration metabolites and were not able to identify these LIUPs reliably. In general, conventional methods, particularly high-resolution analysis, are recommended for peaks with moderate signal-to-noise. The TPU method is recommended for peaks with low signal-to-noise or when compression of spectral data with high fidelity is desirable, such as integration of NMR data into cross-platform studies. PMID- 19551929 TI - High-throughput quantitative polymerase chain reaction in picoliter droplets. AB - Limiting dilution PCR has become an increasingly useful technique for the detection and quantification of rare species in a population, but the limit of detection and accuracy of quantification are largely determined by the number of reactions that can be analyzed. Increased throughput may be achieved by reducing the reaction volume and increasing processivity. We have designed a high throughput microfluidic chip that encapsulates PCR reagents in millions of picoliter droplets in a continuous oil flow. The oil stream conducts the droplets through alternating denaturation and annealing zones, resulting in rapid (55-s cycles) and efficient PCR amplification. Inclusion of fluorescent probes in the PCR reaction mix permits the amplification process to be monitored within individual droplets at specific locations within the microfluidic chip. We show that amplification of a 245-bp adenovirus product can be detected and quantified in 35 min at starting template concentrations as low as 1 template molecule/167 droplets (0.003 pg/microL). The frequencies of positive reactions over a range of template concentrations agree closely with the frequencies predicted by Poisson statistics, demonstrating both the accuracy and sensitivity of this platform for limiting dilution and digital PCR applications. PMID- 19551930 TI - Application of photonic crystal enhanced fluorescence to a cytokine immunoassay. AB - Photonic crystal surfaces are demonstrated as a means for enhancing the detection sensitivity and resolution for assays that use a fluorescent tag to quantify the concentration of an analyte protein molecule in a liquid test sample. Computer modeling of the spatial distribution of resonantly coupled electromagnetic fields on the photonic crystal surface are used to estimate the magnitude of enhancement factor compared to performing the same fluorescent assay on a plain glass surface, and the photonic crystal structure is fabricated and tested to experimentally verify the performance using a sandwich immunoassay for the protein tumor necrosis factor-alpha (TNFalpha). The demonstrated photonic crystal fabrication method utilizes a nanoreplica molding technique that allows for large area inexpensive fabrication of the structure in a format that is compatible with confocal microarray laser scanners. The signal-to-noise ratio for fluorescent spots on the photonic crystal is increased by at least 5-fold relative to the glass slide, allowing a TNF-alpha concentration of 1.6 pg/mL to be distinguished from noise on a photonic crystal surface. In addition, the minimum quantitative limit of detection on the photonic crystal surface is one-third the limit on the glass slide--a decrease from 18 to 6 pg/mL. The increased performance of the immunoassay allows for more accurate quantitation of physiologically relevant concentrations of TNF-alpha in a protein microarray format that can be expanded to multiple cytokines. PMID- 19551931 TI - Design of an oligonucleotide-incorporated nonfouling surface and its application in electrochemical DNA sensors for highly sensitive and sequence-specific detection of target DNA. AB - In this work, we report a novel electrochemical DNA sensor based on a nonfouling monolayer structure self-assembled at gold surfaces. Self-assembled monolayers (SAMs) with oligo(ethylene glycol) (OEG)-terminated thiols are known to be highly protein-resistant and effectively repel nonspecific adsorption. We found that a mixed SAM structure incorporating thiolated oligonucleotides and OEG thiols (SH DNA/OEG) exhibited the similar non-fouling feature. More importantly, it allowed facile electron transfer across the monolayer and thus was fully compatible with electrochemical detection. On the basis of this SH-DNA/OEG platform, we developed a sandwich-type electrochemical sensor for the sequence-specific detection of DNA targets. This sensor was able to detect as little as 1 pM target DNA even in the presence of complicated biological fluids such as human serum. We also employed this sensor to directly detect a polymerase chain reaction (PCR) amplicon from the genomic DNA of Escherichia coli K12, which led to a very low detect limit of 60 fg (approximately 10 copies). PMID- 19551932 TI - Development of a selective ESI-MS derivatization reagent: synthesis and optimization for the analysis of aldehydes in biological mixtures. AB - In LC-MS, derivatization is primarily used to improve ionization characteristics, especially for analytes that are not (efficiently) ionized by ESI or APCI such as aldehydes, sugars, and steroids. Derivatization strategies are then directed at the incorporation of a group with a permanent charge. A compound class that typically requires derivatization prior to LC-MS is the group of small aliphatic aldehydes that are, for instance, analyzed as the key biomarkers for lipid peroxidation in organisms. Here we report the development of a new tailor-made, highly sensitive, and selective derivatization agent 4-(2 (trimethylammonio)ethoxy)benzenaminium halide (4-APC) for the quantification of aldehydes in biological matrixes with positive ESI-MS/ MS without additional extraction procedures. 4-APC possesses an aniline moiety for a fast selective reaction with aliphatic aldehydes as well as a quaternary ammonium group for improved MS sensitivity. The derivatization reaction is a convenient one-pot reaction at a mild pH (5.7) and temperature (10 degrees C). As a result, an in vial derivatization can be performed before analysis with an LC-MS/MS system. All aldehydes are derivatized within 30 min to a plateau, except malondialdehyde, which requires 300 min to reach a plateau. All derivatized aldehydes are stable for at least 35 h. Linearity was established between 10 and 500 nM and the limits of detection were in the 3-33 nM range for the aldehyde derivatives. Furthermore, the chosen design of these structures allows tandem MS to be used to monitor the typical losses of 59 and 87 from aldehyde derivatives, thereby enabling screening for aldehydes. Finally, of all aldehydes, pentanal and hexanal were detected at elevated levels in pooled healthy human urine samples. PMID- 19551933 TI - A new dynamic in mass spectral imaging of single biological cells. AB - Time-of-flight secondary ion mass spectrometry (TOF-SIMS) has unique capabilities in the area of high-resolution mass spectrometric imaging of biological samples. The technique offers parallel detection of native and non-native molecules at physiological concentrations with potentially submicrometer spatial resolution. Recent advances in SIMS technology have been focused on generating new ion sources that can in turn be used to eject more intact molecular and biological characteristic species from a sample. The introduction of polyatomic ion beams, particularly C60, for TOF-SIMS analysis has created a whole new application of molecular depth profiling and 3D molecular imaging. However, such analyses, particularly at high lateral resolution, are severely hampered by the accompanying mass spectrometry associated with current TOF-SIMS instruments. Hence, we have developed an instrument that overcomes many of the drawbacks of current TOF-SIMS spectrometers by removing the need to pulse the primary ion beam. The instrument samples the secondary ions using a buncher that feeds into a specially designed time-of-flight analyzer. We have validated this new instrumental concept by analyzing a number of biological samples generating 2D and 3D images showing molecular localization on a subcellular scale, over a practical time frame, while maintaining high mass resolution. We also demonstrate large area mapping and the MS/MS capability of the instrument. PMID- 19551934 TI - Ion mobility spectrometry/mass spectrometry snapshots for assessing the molecular compositions of complex polymeric systems. AB - The synthesis of increasingly complex polymers has created daunting, sometimes insurmountable problems for their chemical analysis. The importance is magnified by outsourcing of production and their use in consumer products, including medical devices and food storage, and therefore requires a new generation of technology for quality assurance. Here, we report capturing subtle differences at the molecular level in complex polymer mixtures nearly instantaneously using a prototype multidimensional ion mobility spectrometry/mass spectrometry spectrometry instrument. Bulk activation/fragmentation strategies reported here provide signatures of structural characteristics that permit effortless recognition of minor differences in blends and copolymers, even as structural isomers and from a quantitative perspective. The data displayed as a pictorial snapshot provide a visual pattern that is sufficiently distinctive that computer aided pattern recognition can be used to address process control and regulatory issues. PMID- 19551935 TI - Preparation of alkanethiolate-functionalized core/shell Fe3O4@Au nanoparticles and its interaction with several typical target molecules. AB - This paper reports the synthesis of alkanethiolate-functionalized core/shell Fe3O4/Au nanoparticles (Fe3O4@Au NPs) that combine the advantages of core/shell magnetic nanoparticles with self-assembled monolayers (SAMs). The alkanethiolates, having carboxylic acid (COOH) and methyl (CH3) terminal groups, can be easily self-assembled on the Fe3O4@Au NPs substrates. The surface composition and structure of 11-mercaptoundecanoic acid (11-MUA) and 1 dodecanethiol (11-DDT) SAMs were characterized in detail using energy-dispersive spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). To further study the surface properties of the SAMs, hydrophobic, ionic, and nonionic organic compounds were selected as probe molecules. The results indicated that the hydrophobic interaction caused strong retention of analytes, whereas the repulsive electrostatic interaction between the negatively charged 11-MUA SAMs surface and the negatively charged probe molecule would lead to the decrease of adsorption affinity. PMID- 19551936 TI - Electrochemical DNA biosensor based on nanoporous gold electrode and multifunctional encoded DNA-Au bio bar codes. AB - A sensitive electrochemical DNA sensor based on nanoporous gold (NPG) electrode and multifunctional encoded Au nanoparticle (AuNP) was developed. The NPG electrode was prepared with a simple dealloying strategy, by which silver was dissoluted from silver/gold alloys in nitric acid, making the active surface area of NPG electrode 9.2 times higher than that of a bare flat one characterized by cyclic voltammetry. A DNA biosensor was fabricated by immobilizing capture probe DNA on the NPG electrode and hybridization with target DNA, which further hybridized with the reporter DNA loaded on the AuNPs. The AuNP contained two kinds of bio bar code DNA, one was complementary to the target DNA, while the other was not, reducing the cross reaction between the targets and reporter DNA on the same AuNP. Electrochemical signals of [Ru(NH3)6]3+ bound to the reporter DNA via electrostatic interactions were measured by chronocoulometry. Taking advantage of dual-amplification effects of the NPG electrode and multifunctional encoded AuNP, this DNA biosensor could detect the DNA target quantitatively, in the range of 8.0 x 10(-17)-1.6 x 10(-12) M, with a limit of detection as low as 28 aM, and exhibited excellent selectivity even for single-mismatched DNA detection. PMID- 19551938 TI - Oligopeptides as mimic of acetylcholinesterase: from the rational design to the application in solid-phase extraction for pesticides. AB - Three different peptides (His-Glu-Pro-Ser, His-Gly-Ser-Ala and Glu-Pro-Ser-Ala) were selected and tested to be used as affinity binding receptors for organophosphate and carbamate pesticides. The peptides were rationally designed by mimicking acetylcholinesterase active site. The simulated binding energy of the three tetrapeptides versus one model of organophosphate (paraoxon) and one of carbamate (carbaryl) pesticide was calculated; a good correlation between shape designed and binding score was obtained. The binding properties of the peptide pesticide interaction were studied following the variation of UV-visible spectra in different solvents. The binding constants in water, which were nicely correlated with computational data, ranged from 506 (+/-115) to 36(+/-2) M(-1). All the peptides had a 5-fold decrease in binding by changing solvent, going from water to less polar ethanol. The binding affinity suggested the use of these ligands as a preanalytical tool in extraction cartridges. The tetrapeptides efficiency was tested linking the peptides to two different supports. The cartridges prepared using His-Glu-Pro-Ser sequence was, as predicted, able to bind paraoxon and carbaryl with recovery values in the 72-88% range at pH 4.5. Intercolumn, interday RSD was in the 4-7% range. The columns were used for 80 cycles before losing retention ability. PMID- 19551937 TI - Assignment of disulfide-linked peptides using automatic a1 ion recognition. AB - We present a novel approach for the assignment of peptides containing disulfide linkages. Dimethyl labeling is introduced to generate labeled peptides which exhibit enhanced a1 ion signals during MS/MS fragmentation. For disulfide-linked peptides, multiple a1 ions can be observed due to multiple N-termini. This distinct feature allows sieving out the disulfide-linked peptides; meanwhile, the N-terminal amino acids can be identified. With such information, the number of possible peptide combinations involved in a disulfide bond dramatically narrows down. Furthermore, we developed a computational algorithm to perform target a1 ion screening followed by molecular weight matching of cysteine-containing peptides with specific amino acids at the N-termini. Once the protein sequence and the peak list from a LC-MS/MS survey scan of labeled peptides are imported, the identities of disulfide-linked peptides can be readily obtained. The presented approach is simple and straightforward, offering a valuable tool for protein structural characterization. PMID- 19551939 TI - Memory effects in compound-specific D/H analysis by gas chromatography/pyrolysis/isotope-ratio mass spectrometry. AB - Compound-specific analyses of lipid D/H ratios often encounter ranges of 300 per thousand or more, and experiments using D-enriched water to study fractionations often extend the range up to 1000 per thousand. Here we show that for such large dynamic ranges in D/H ratio, isotopic "memory" between adjacent peaks can be significant. Memory effects have not been previously reported for GC/P/IRMS systems but can have a significant impact on many measurements, even those exploring only natural-abundance variations in D/H. To quantitatively evaluate these effects, we synthesized two series of organic standards with deltaD values varying from -230 to +800 per thousand. We then analyzed chromatograms in which analyte deltaD values, retention times, or relative abundances were independently varied. For two sequential GC peaks, isotopic memory is measured to be typically 2-4% of the difference in deltaD values between the two. Roughly half of this effect can be attributed to unknown processes within the GC itself, and the other half to surface adsorption processes in the pyrolytic conversion of analytes to H2. Isotopic memory increases with decreasing time separation between peaks, with decreasing analyte abundance, and with increasing age of pyrolysis reactors. A simple numerical model that simulates dynamic adsorption of H2 on pyrolytic carbon can reproduce many aspects of the experimental data, suggesting that this is likely to be an important mechanism in isotopic memory. Several steps to mitigate memory effects in routine analyses are suggested. PMID- 19551940 TI - Obtaining Nernstian response of a Ca(2+)-selective electrode in a broad concentration range by tuned galvanostatic polarization. AB - Linear Nernstian response is obtained for a neutral ionophore-based Ca(2+) selective electrode down to 10(-10) M CaCl2 by means of galvanostatic polarization. The densities of the applied cathodic current were tuned for particular concentrations of Ca2+. The procedure included recording the potential at zero current, followed by measurements when current is passed through the electrode, and then again at zero current. The respective chronopotentiometric curves included negative ohmic drop immediately after turning the current on, the polarization domain, and positive ohmic drop when the current was turned off, followed with the relaxation domain. The potentials immediately after the positive ohmic drops were used as analytical signals. These potentials make a straight line with Nernstian slope when currents are tuned (optimized) for each particular concentration. An iteration procedure is proposed which allows for simultaneous optimization of the current density and accessing analyte concentration in the sample. PMID- 19551941 TI - Diethylation labeling combined with UPLC/MS/MS for simultaneous determination of a panel of monoamine neurotransmitters in rat prefrontal cortex microdialysates. AB - The primary challenge associated with the development of an LC/MS/MS-based assay for simultaneous determination of biogenic monoamine neurotransmitters such as norepinephrine (NE), dopamine (DA), serotonin (5-HT), and normetanephrine (NM) in rat brain microdialysates is to improve detection sensitivity. In this work, a UPLC/ MS/MS-based method combined with a diethyl labeling technique was developed for simultaneous determination of a panel of monoamines in rat prefrontal cortex microdialysates. The chromatographic run time is 3.5 min/ sample. The limits of detection of the UPLC/MS/MS-based method for NE, DA, 5-HT/ and NM, with/without diethyl labeling of monoamines, are 0.005/0.4 (30/2367 pM), 0.005/0.1 (33/653 pM), 0.005/0.2 (28/1136 pM), and 0.002/0.2 ng/mL (11/1092 pM), respectively. Diethyl labeling of amino groups of monoamines affords 20-100 times increased detection sensitivity of corresponding native monoamines during the UPLC/MS/MS analysis. This could result from the following: (1) improved fragmentation patterns; (2) increased hydrophobicity and concomitantly increased ionization efficiency in ESI MS and MS/MS analysis; (3) reduced matrix interference. This labeling reaction employs a commercially available reagent, acetaldehyde-d4, to label the amine groups on the monoamines via reductive amination. It is also simple, fast (approximately 25-min reaction time), specific, and quantitative under mild reaction conditions. Data are also presented from the application of this assay to monitor the drug-induced changes of monoamine concentrations in rat prefrontal cortex microdialysate samples followed by administration of SKF 81297, a selective D1 dopamine receptor agonist known to elevate the extracellular level of the neurotransmitters DA and NE in the central nervous system. PMID- 19551942 TI - Precise isotopic analysis of Mo in seawater using multiple collector-inductively coupled mass spectrometry coupled with a chelating resin column preconcentration method. AB - It is widely recognized that the natural isotopic variation of Mo can provide crucial information about the geochemical circulation of Mo, and the ocean is an important reservoir of Mo. To obtain precise isotopic data on Mo in seawater samples using multiple collector-inductively coupled plasma mass spectrometry (MC ICPMS), we have developed a preconcentration technique using 8-hydroxyquinoline bonded covalently to a vinyl polymer resin (TSK-8HQ). By optimizing the procedure, Mo in seawater could be effectively separated from matrix elements such as alkali, alkaline earth, and transition metals. With this technique, even with a 50-fold enrichment factor, the changes in the 98Mo/95Mo ratio during preconcentration were smaller than twice the standard deviation (SD) in this study. Mass discrimination of Mo isotopes during the measurement was externally corrected for by normalizing 86Sr/88Sr to 0.1194 using an exponential law. We evaluated delta98/95Mo to a precision of +/- 0.08 per thousand (+/-2 SD); this value was found to be less than one-third of previous reported values. Moreover, we were able to determine an accurate ratio for every pair of stable Mo isotopes, which was impossible with previous methods owing to the isobaric interference from the external elements (Zr and Ru). In this study, delta92/98Mo in seawater was first determined so that it had the smallest relative error. We applied the proposed method to four kinds of seawater samples. The Mo compositions were constant among them, with average delta98/95Mo and delta92/98Mo values of 2.45 +/ 0.11 and -4.94 +/- 0.09 per thousand (+/-2 SD), respectively. Our data indicate that seawater is enriched in heavy Mo isotopes than previously reported. PMID- 19551943 TI - Reversed-phase LC/MS method for polyphosphoinositide analyses: changes in molecular species levels during epidermal growth factor activation in A431 cells. AB - In studies on lipid metabolomics, liquid chromatography/ mass spectrometry (LC/MS) is a robust and popular technique. Although effective reversed-phase (RP) LC/ MS methods enabling the separation of phospholipid molecular species have been developed, RPLC methods to analyze phosphatidylinositol phosphates (PIPs) have not been reported. In this study, we developed conditions suitable for PIP analysis. Coupled with (diethylamino)ethyl (DEAE)-cellulose pretreatment, at least 1 pmol each of phosphatidylinositol monophosphates (PIP1), bisphosphates (PIP2), and triphosphates standards per approximately 6 x 10(6) cultured cells could be measured. Using these methods, we detected elevated concentrations of more than 12 PIP1 species in epidermal growth factor (EGF)-stimulated A431 cells, a human epidermoid carcinoma cell line. The PIP2 species detected were not elevated after stimulation. We also detected EGF-induced increases in the levels of several phosphatidic acid species using another set of methods. Our method sensitively determined PIPs within a biological sample and is thus suitable for analysis of phoisphoiniositide metabolism. PMID- 19551944 TI - Calibration-free quantitative application of in situ Raman spectroscopy to a crystallization process. AB - In this work, in situ Raman spectroscopy is applied in a quantitative manner to crystal suspensions during crystallization in order to monitor a solvent-mediated polymorph transformation without the use of a calibration model. Assuming a linear dependency of the Raman signal intensity on the solute and on the solid concentrations of both solid-state forms, the measured time-resolved Raman spectra are fitted directly using a detailed model that describes the time evolution of the process. The applicability of this novel method is demonstrated thoroughly through the application to synthetic data of unseeded and seeded transformations as well as to various seeded polymorph transformation experiments. The resulting concentration profiles show a good agreement with the concentrations obtained by a multivariate calibration model. Additionally, the estimated kinetic parameters are compared to parameters obtained through fitting the solid-phase composition profiles that result from the calibration. The discrepancy between the estimated model parameters is small, and essentially the same descriptive process model is obtained. However, by fitting the time-resolved Raman spectra directly, a significant amount of calibration effort can be avoided. PMID- 19551945 TI - Integration of microdialysis sampling and microchip electrophoresis with electrochemical detection. AB - Here we describe the fabrication, optimization, and application of a microfluidic device that integrates microdialysis (MD) sampling, microchip electrophoresis (ME), and electrochemical detection (EC). The manner in which the chip is produced is reproducible and enables the fixed alignment of the MD/ME and ME/EC interfaces. Poly(dimethylsiloxane) (PDMS)-based valves were used for the discrete injection of sample from the hydrodynamic MD dialysate stream into a separation channel for analysis with ME. To enable the integration of ME with EC detection, a palladium decoupler was used to isolate the high voltages associated with electrophoresis from micrometer-sized carbon ink detection electrodes. Optimization of the ME/EC interface was needed to allow the use of biologically appropriate perfusate buffers containing high salt content. This optimization included changes in the fabrication procedure, increases in the decoupler surface area, and a programmed voltage shutoff. The ability of the MD/ME/EC system to sample a biological system was demonstrated by using a linear probe to monitor the stimulated release of dopamine from a confluent layer of PC 12 cells. To our knowledge, this is the first report of a microchip-based system that couples microdialysis sampling with microchip electrophoresis and electrochemical detection. PMID- 19551946 TI - Matrix solid-phase dispersion as a sample pretreatment for the speciation of arsenic in seafood products. AB - Matrix solid-phase dispersion (MSPD) has been applied to extract arsenical species (arsenite, As(III); arsenate, As(V); monomethylarsonic acid; dimethylarsinic acid, DMA; arsenobetaine, AsB; and arsenocholine) from seafood products. High-performance liquid chromatography coupled to inductively coupled plasma-mass spectrometry was used to separate and detect all arsenic species. Variables affecting MSPD, such as the solid support material (dispersing agent), solid support mass/sample mass ratio, elution solvent composition, and elution solvent volume, have been fully evaluated. Quantitative recoveries for inorganic and organic arsenic species have been obtained when using diatomaceous earth or octadecyl-functionalized silica gel (C18) as a solid support material, with a solid support mass/sample mass ratio of 7.0. Elution of arsenical compounds has been assessed using 10 mL of 50/50 methanol/ultrapure water as an elution solvent. The MSPD method has been found precise, with RSDs of approximately 9% for As(III), DMA, and As(V) and 3% for AsB. The developed procedure has been tested by analyzing different certified reference materials of marine origin such as DORM-2 and BCR 627, which offer certified contents for some arsenic species. The method has been also applied to assess arsenic speciation in different mollusks, cold water fishes, and white fishes. PMID- 19551947 TI - Urinary metabolite quantification employing 2D NMR spectroscopy. AB - Two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy is a fairly novel method for the quantification of metabolites in biological fluids and tissue extracts. We show in this contribution that, compared to 1D 1H spectra, superior quantification of human urinary metabolites is obtained from 2D 1H-13C heteronuclear single-quantum correlation (HSQC) spectra measured at natural abundance. This was accomplished by the generation of separate calibration curves for the different 2D HSQC signals of each metabolite. Lower limits of detection were in the low to mid micromolar range and were generally the lower the greater the number of methyl groups contained in an analyte. The quantitative 2D NMR data obtained for a selected set of 17 urinary metabolites were compared to those obtained independently by means of gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry of amino acids and hippurate as well as enzymatic and colorimetric measurements of creatinine. As a typical application, 2D-NMR was used for the investigation of urine from patients with inborn errors of metabolism. PMID- 19551949 TI - Instance based algorithm for posterior probability calculation by target-decoy strategy to improve protein identifications. AB - The target-decoy database search strategy is often applied to determine the global false-discovery rate (FDR) of peptide identifications in proteome research. However, the confidence of individual peptide identification is typically not determined. In this study, we introduced an approach for the calculation of posterior probability of individual peptide identification from the "local false-discovery rate" (local FDR), which is also determined based on a target-decoy database search. The peptide identification scores output by the database search algorithm were weighted by their discriminating power using a Shannon information entropy based strategy. Then the local FDR of a peptide identification was calculated based on the fraction of decoy identifications among its nearest neighbors within a small space defined by these weighted scores. It was demonstrated that the calculated probability matched the actual probability precisely, and it provided powerful discriminating performance between true positive and false positive identifications. Hence, the sensitivity for peptide identification as well as protein identification was significantly improved when the calculated probability was used to process different proteome data sets. As an instance based strategy, this algorithm provides a safe way for the posterior probability calculation and should work well for data sets with different characteristics. PMID- 19551951 TI - Analysis of hybrid silica materials with the aid of conventional NMR and GC/MS. AB - Two simple and straightforward procedures for determining the organic content of hybrid silica materials by means of conventional NMR and GC/MS techniques are described. The methods involve dissolving the hybrid material either in a concentrated solution of sodium hydroxide in deuterated water containing a suitable reference or in a solution of hydrogen fluoride in water and extracting with methylene chloride. These methods constitute useful routine techniques for obtaining immediate information concerning both the amount and chemical composition of the organic compounds on the silica surface. PMID- 19551948 TI - Continuous and quantitative delivery of molecules into individual cells with a diffusional microburet. AB - Direct delivery of molecules into the cytosol of live cells is required in many areas of biology and clinical research. Molecules of interest include indicator dyes, biomolecules, and pharmacological agents. In this work we describe continuous delivery of molecules into single cells using a diffusional microburet, DMB. The DMB is a pulled glass micropipette with a fine tip that contains a microscopic plug made of a hydrogel such as agar or polyacrylamide. This plug prevents flow but allows diffusive delivery of the molecule of interest from the DMB body into the cytosol, driven by its concentration gradient. This leads to a scheme of sustained intracellular dosing that is highly reproducible and quantifiable yet does not require the addition of solution volume to the cell. Potential loss of biomolecules from the cytosol through the plug of the DMB can be greatly reduced by proper choice of the pore size and tortuosity of the hydrogel in the DMB tip. The intracellular concentration of fluorescent molecules during delivery can be obtained calibration free. In this work we demonstrate dosing of Lucifer Yellow CH, LY, a charged fluorescent dye, into individual a7r5 vascular smooth muscle cells with a DMB. New types of quantitative analytical experiments on single live cells that the DMB technology enables are titration of intracellular ions and ligands, binding sites, and efflux pathways such as those that are involved in drug resistance. PMID- 19551950 TI - Overcoming matrix effects in liquid chromatography-mass spectrometry. AB - A major limitation in quantitative analysis with electrospray ionization mass spectrometry (ESI-MS) is represented by the so-called matrix effects in which the matrix coextracted with the analytes can alter the signal response, causing either suppression or enhancement, resulting in poor analytical accuracy, linearity, and reproducibility. In the direct electron ionization liquid chromatography-mass spectrometry (direct-EI LC-MS) interface the ionization process is based on electron impact ionization, and it occurs in the gas phase and is not influenced by coeluted matrix compounds. In this work we quantitatively evaluated matrix effects on enriched environmental and biological samples, with different extraction procedures, using ESI and direct-EI LC-MS. As expected, the samples analyzed with direct-EI were not affected by matrix composition, whereas with ESI we observed either signal suppression or enhancement, depending on the sample nature. PMID- 19551952 TI - Multiplex ligation-dependent probe amplification technique for copy number analysis on small amounts of DNA material. AB - The multiplex ligation-dependent probe amplification (MLPA) technique is a sensitive technique for relative quantification of up to 50 different nucleic acid sequences in a single reaction, and the technique is routinely used for copy number analysis in various syndromes and diseases. The aim of the study was to exploit the potential of MLPA when the DNA material is limited. The DNA concentration required in standard MLPA analysis is not attainable from dried blood spot samples (DBSS) often used in neonatal screening programs. A novel design of MLPA probes has been developed to permit for MLPA analysis on small amounts of DNA. Six patients with congenital adrenal hyperplasia (CAH) were used in this study. DNA was extracted from both whole blood and DBSS and subjected to MLPA analysis using normal and modified probes. Results were analyzed using GeneMarker and manual Excel analysis. A total number of 792 ligation events were analyzed. In DNA extracted from dried blood spot samples, 99.1% of the results were accurate compared to 99.9% of the results obtained in DNA from whole blood samples. This study clearly demonstrates that MLPA reactions with modified probes are successful and reliable with DNA concentrations down to 0.3 ng/microL (1.6 ng total). This broadens the diagnostic perspectives of samples of DBSS allowing for copy number variation analysis in general and particularly testing for CAH. PMID- 19551953 TI - Response to comment on "Preparation of superparamagnetic ribonuclease A surface imprinted submicrometer particles for protein recognition in aqueous media". PMID- 19551954 TI - Take care when charting patients. PMID- 19551958 TI - [Antibodies to enzymes of purine metabolism as a factor of gastrointestinal tract lesions in systemic scleroderma]. AB - Antibodies to Adenosine Deaminase (AD) and Guanin Deaminase (GDA) were founded in sera of systemic sclerosis (SS) patients, their concentration positive correlated with activity of disease. Increased levels of antibodies to AD and GDA correlated with the severity of gastrointestinal tract injury (especially pancreatitis and hepatitis). It is shown that level of antibodies to AD and GDA differs reliable from contents of these antibodies, in sera of healthy persons. It is shown that AD enzymatic activity was decreased, while activity of GDA was increased. The antibodies to enzymes may be one of the possible causes of change of enzymes activity in sera of SS patients. The method of immunoenzyme determination of level of antibodies to AD, G on the basis of immobilized form of magnet sorbent was developed for immune diagnostics of SS. PMID- 19551959 TI - [Dynamics of pro-and anti-inflammatory cytokines at patients with combination of chronic opisthorchiasis with Helicobacter pylori-associated gastritis]. AB - The aim of our study was to estimate the cytokins changes (TNF alpha, IL-1 beta, IL-8, IL-4) in patients with combination of chronic opisthorhosis and Helicobacter pylori associated gastritis and to estimate degelmintizational therapy with biltricid on cytokine levels. The 55 patients were investigated and divided on two groups: isolated opisthorhosis -- 20 patients and opisthorhosis in combination with Helicobacter pylori associated gastritis -- 35 patients. In both groups of patients was estimated the increase of TNF alpha, IL-1 beta, IL-8, IL-4 levels. The treatment with biltricid allow to decrease the TNF alpha, IL-1beta, IL-8 levels and to increase IL-4 level, especially in patients with combination of chronic opisthorhosis and Helicobacter pylori associated gastritis. PMID- 19551960 TI - [Effectiveness of panzytrat--modern physiological enzyme preparation in complex therapy of pancreatic exocrine secretory insufficiency in cholelithiasis]. AB - In the article the analysis of the survey with 102 patients with gallstone disease involved, 68 of whom underwent cholecystectomy and 34 were treated conservatively, is made. The content of fecal elastase 1 in stool was estimated for diagnostics of exocrine enzyme insufficiency of pancreas by immune-enzyme analysis. It was stated that 90% of patients possess secondary exocrine insufficiency of pancreas in case of gallstone disease. It is the result of complex metabolic liver abnormalities, portal and mesenterial haemodynamics, dysbiosis of large intestine which are the components of a syndrom of maldigestion and appear during gallstone disease progressing long time before hospitalization. Cholecystectomy doesn't eliminate enzyme insufficiency of pancreas. The effectiveness of using new physiological enzymatic drug Panzytrat in a complex therapy of a syndrom of maldigestion in case of gallstone disease is shown. PMID- 19551961 TI - [The role of public health in the implementation of the demographic policy concept on the regional level]. AB - The major trends of the social demographic processes in the Novgorodskaya oblast are characterized. The activities to increase the level of natality rate and to enhance the population reproductive health are proposed. The set of measures is developed to decrease the mortality of population of able-bodied age. The prospective results of demographic policy applied in the Novgorodskaya oblast up to 2011 are exposed. The role of public health in the enhancement of demographic situation on the regional level is specified. PMID- 19551963 TI - [The characteristics of children disability on the Northern Territories]. AB - The materials related to the regional characteristics of health formation in the disabled children are presented. The corresponding evaluation was made using the conventional and new methodological approach based on the so-called "three dimensional concept". The obtained data is the initial base for developing and implementation of the integrated programs of the medical social, psychological pedagogical and social economical rehabilitation of the disabled children and their families. PMID- 19551962 TI - [The medical activity of population resident in rural area]. AB - The medical social study was carried out to investigate the degree of the prevalence of medical activity among the inhabitants of rural area. The attitude of the respondents to the preventive medical check-up was determined. The timeliness of having recurrence to medical care, the performing of physician prescriptions, the independent decision-making related to the medical treatment and the degree of awareness of healthy life-style have been explored. The study was organized in the large rural region of Chelyabinsk oblast. The study sampling consisted of 1740 respondents. The lower level of medical activity among rural inhabitants was revealed. The necessity to enhance the organization of medical institutions functioning related to sanitary hygienic education of rural population and to elaborate the medical care organization. PMID- 19551964 TI - [The medical social characteristics of patients hospitalized in the traumatic surgery department]. AB - The medical social characteristics of injured patients hospitalized in the traumatic surgery department of municipal hospital. The study results permitted to develop the organizational functional model of injury prevention management. The pattern presuppose the implementation of complex of activities considering the options of target population groups and measures realization period based on the regular occurrences revealed in the study. PMID- 19551965 TI - [The application of the quantitative analysis technique in the evaluation of the effectiveness of treatment and prevention of diseases]. AB - The optimization model of cost effectiveness analysis as applied to select the medical diagnostic and medical prevention programs with the view of increasing the losses of population health capital. The program implementation of the model and recommendations on its practical use at management decision-making in public health are exposed. PMID- 19551966 TI - [The medical organizational resources of fertility promotion in the Republic of Belarus]. AB - The assessment of youth reproductive attitudes and the identification of the medical organizational opportunities for fertility promotion in the Republic of Belarus is considered. Relying on the data of the sociological survey of representative youth sample of national level it is established that both the reproductive attitudes and reproductive behavior of Belarus youth population does not correspond to the demographic criteria of effective population reproduction. It is revealed that the enhancement of reproductive attitudes of young women is possible in the conditions of increase of the rate of first pregnancy planning. The economic benefit due to possible implementation of this uncovered resource is evaluated. PMID- 19551967 TI - [Analysis of expression of heavy myosin chains during in vitro differentiation of satellite cells and myoblasts derived from rat skeletal muscles]. AB - Differentiation of cultured myogenic progenitor cells (satellite cells and mononucleated myoblasts) derived from hindlimb muscles of rat embryos and newborn animals was studied. Immunocytochemical methods and PCR analysis revealed expression of heavy myosin chains at the earliest stages of myogenesis (in mononucleated myoblasts). Expression of the gene encoding the embryonic form of myosin and a low level of expression of the gene encoding perinatal myosin in cultured progenitor cells derived from embryonic muscles was detected by PCR. Cells derived from muscles of newborn animals also expressed these two myosin forms, though at a lower level. The progenitor cells derived from muscles of rat embryos and newborn animals were found to express myosin 2a, which is characteristic of fast-twitch definitive muscle fibers. PMID- 19551968 TI - [Freezing tolerance of winter wheat plants depends on adaptation of photosynthesis and respiration in different time intervals]. AB - This study is devoted to CO2 gas exchange (true photosynthesis at light saturation (P), dark respiration (R), and P/R ratio) in vegetating and cold hardened winter wheat (Triticum aestivum L.) plants (cultivar Mironovskaya 808) in relation to their freezing tolerance. Under natural cultivation conditions, freezing tolerance of plants depended on adaptive changes in the shape of P and R curves in the temperature range from 20 to -2 degrees C. These changes, induced by cold hardening and treatment of plants with the photosynthesis inhibitor diuron, were observed within month and week ranges. Under laboratory conditions, the P/R ratio in vegetating plants increased three times within an hour range as the temperature decreased from 22 to 0 degree C. The P/R ratio also decreased within a minute range as a result of partial inhibition of photosynthesis with diuron and immediately decreased when CO2 concentration in the air was reduced from 419 to 0 microl/l. The P/R ratio decreased primarily at the expense of a decrease in P. The decrease in P/R was more pronounced at low temperatures, indicating variability of low-temperature tolerance of photosynthesis within a minute range. The possibility of plant adaptation to nonsimultaneous temperature changes under natural conditions via adaptive changes in temperature tolerance of the photosynthetic apparatus is discussed. PMID- 19551969 TI - Solvation effects in the quartz crystal microbalance with dissipation monitoring response to biomolecular adsorption. A phenomenological approach. AB - Quartz crystal microbalance with dissipation monitoring (QCM-D) has become a popular tool to investigate biomolecular adsorption phenomena at surfaces. In contrast to optical mass-sensitive techniques, which commonly detect the adsorbed nonhydrated mass, the mechanically coupled mass measured by QCM-D includes a significant amount of water. A mechanistic and quantitative picture of how the surrounding liquid couples to the deposited solutes has so far been elusive for apparently simple phenomena like the random adsorption of nanometer-sized particles on a planar surface. Using a setup that enables simultaneous measurements by reflectometry and QCM-D on the same support, we have quantified the variations in coupled water, as sensed by the QCM frequency response, as a function of coverage for the formation of monolayers of globular proteins, virus particles, and small unilamellar vesicles. We found a close-to-linear relationship between the surface coverage and the relative contribution of water to the frequency response for these adsorption scenarios. The experimental hydration curves could be reproduced quantitatively using a theoretical model that assigns a pyramid-shaped hydration coat to each adsorbed particle and that accounts for the random distribution of adsorbents on the surface. This simple model fits the experimental data well and provides insight into the parameters that affect hydration. PMID- 19551970 TI - Electroacoustic polymer microchip as an alternative to quartz crystal microbalance for biosensor development. AB - Laser photoablation of poly(ethylene terephthalate) (PET), a flexible dielectric organic polymer, was used to design an acoustic miniaturized DNA biosensor. The microchip device includes a 100-microm-thick PET layer, with two microband electrodes patterned in photoablated microchannels on one side and a depressed photoablated disk decorated by gold sputtered layer on the other side. Upon application of an electric signal between the two electrodes, an electroacoustic resonance phenomenon at approximately 30 MHz was established through the microelectrodes/PET/ gold layer interface. The electroacoustic resonance response was fitted with a series RLC motional arm in parallel with a static Co arm of a Buttlerworth-Van Dyke equivalent circuit: admittance spectra recorded after successive cycles of DNA hybridization on the gold surface showed reproducible changes on R, L, and C parameters. The same hybridizations runs were performed concomitantly on a 27-MHz (9 MHz, third overtone) quartz crystal microbalance in order to validate the PET device developed for bioanalysis applications. The electroacoustic PET device, approximately 100 times smaller than a microbalance quartz crystal, is interesting for the large-scale integration of acoustic sensors in biochips. PMID- 19551971 TI - On-line digestion system for protein characterization and proteome analysis. AB - An efficient on-line digestion system that reduces the number of sample manipulation steps has been demonstrated for high-throughput proteomics. By incorporating a pressurized sample loop into a liquid chromatography-based separation system, both sample and enzyme (e.g., trypsin) can be simultaneously introduced to produce a complete, yet rapid digestion. Both standard proteins and a complex Shewanella oneidensis global protein extract were digested and analyzed using the automated online pressurized digestion system coupled to an ion mobility time-of-flight mass spectrometer, an ion trap mass spectrometer, or both. The system denatured, digested, and separated product peptides in a manner of minutes, making it amenable to on-line high-throughput applications. In addition to simplifying and expediting sample processing, the system was easy to implement and no cross-contamination was observed among samples. As a result, the online digestion system offers a powerful approach for high-throughput screening of proteins that could prove valuable in biochemical research (rapid screening of protein-based drugs). PMID- 19551972 TI - Comparing capillary electrophoresis-mass spectrometry fingerprints of urine samples obtained after intake of coffee, tea, or water. AB - Metabolomic fingerprinting is a growing strategy for characterizing complex biological samples without detailed prior knowledge about the metabolic system. A two-way analysis system with liquid separation and mass spectrometric detection provides detail-rich data suitable for such fingerprints. As a model study, human urine samples, obtained after intake of coffee, tea, or water, were analyzed with capillary electrophoresis electrospray ionization time-of-flight mass spectrometry (CE-ESI-TOF-MS). In-house-developed software (in Matlab) was utilized to manage and explore the large amount of data acquired (230 CE-MS runs, each with 50-100 million nonzero data points). After baseline and noise reduction, followed by suitable binning in time and m/z, the data sets comprised 9 and 14 million data points in negative and positive ESI mode, respectively. Finally, a signal threshold was applied, further reducing the number to about 100 000 data points per data set. A set of interactive exploratory tools, utilizing principal component analysis (PCA) and analysis of variance (ANOVA) results based on a general linear model, facilitated visual interpretation with score plots (for group assessment) and differential fingerprints (for "hot spot" detection). In the model study highly significant differences due to beverage intake were obtained among the 10 first principal components (p < 10(-6) for two of the components in both ESI modes). Especially, the contrasts between "coffee" and "tea or water" indicated several "hot spots" with highly elevated intensities (e.g., for uncharged masses 93, 94, 109, 119, 123, 132, 148, 169, 178, 187, 190, and 193) suitable for further analysis, for example, with tandem MS. PMID- 19551973 TI - Determination of pesticide residues in fruit-based soft drinks. AB - Here we report the first worldwide reconnaissance study of the presence and occurrence of pesticides in fruit-based soft drinks. While there are strict regulations and exhaustive controls for pesticides in fruits, vegetables, and drinking water, scarce attention has been paid to highly consumed derivate products, which may contain these commodities as ingredients. In the case of the fruit-based soft drinks industry, there are no clear regulations, relating to pesticides, which address them, even when there is significant consumption in vulnerable groups such as children. In this work, we have developed a screening method to search automatically for up to 100 pesticides in fruit-based soft drinks extracts based on the application of liquid chromatography-electrospray time-of-flight mass spectrometry (LC-TOF MS). The sample extracts injected were obtained by a preliminary sample treatment step based on solid-phase extraction using hydrophilic-lipophilic balanced polymer-based reverse phase cartridges and methanol as eluting solvent. Subsequent identification, confirmation, and quantitation were carried out by LC-TOF MS analysis: the confirmation of the target species was based on retention time matching and accurate mass measurements of protonated molecules ([M + H]+) and fragment ions (obtaining accuracy errors typically lower than 2 ppm). With the proposed method, we measured over 100 fruit-based soft drink samples, purchased from 15 different countries from companies with brands distributed worldwide and found relatively large concentration levels of pesticides in most of the samples analyzed. The concentration levels detected were of the micrograms per liter level, low when considering the European maximum residue levels (MRLs) set for fruits but very high (i.e., 300 times) when considering the MRLs for drinking or bottled water. The detected pesticides (carbendazim, thiabendazole, imazalil and its main degradate, prochloraz and its main degradate, malathion, and iprodione) are mainly those applied to crops in the final stages of production (postharvest treatment), some of them contain chlorine atoms in their structures. Therefore, steps should be taken with the aim of removing any traces of pesticides in these products, in order to avoid this source of pesticide exposure on the consumer, particularly on vulnerable groups with higher exposure, such as children. PMID- 19551974 TI - Effect of sample heterogeneity on the interpretation of quartz crystal microbalance data: impurity effects. AB - Quartz crystal microbalance (QCM or QCM-D) has become instrumental in life sciences and biosensor research. It is routinely and successfully used for monitoring interfacial processes, such as protein adsorption and conformational changes in the protein-adsorbed films, liposome-surface interactions and supported bilayer formation, and in the development of biosensor platforms. However, quantitative interpretation of QCM data from biological interfaces studied in liquid remains challenging. In vacuum, the so-called Sauerbrey relationship is routinely used to relate QCM frequency shifts due to the adsorbed layer to the mass of the adsorbed layer. Deviations from Sauerbrey relationship are typically observed when studying soft interfaces in liquids; these are interpreted in terms of layer viscoelastic properties. In this study, we develop and use a combined atomic force microscopy (AFM)-QCM setup to investigate the adsorption of protein ferritin on the surface of gold. First, we find that deviations from the Sauerbrey relationship in this system originate almost entirely from the heterogeneity of the protein films caused by the presence of impurities. Second, relying on the ability of AFM to visualize single ferritin molecules adsorbed on the surface, we find that the frequency shifts determined by QCM are not linearly related to the protein surface coverage. PMID- 19551975 TI - Identification of viruses using microfluidic protein profiling and Bayesian classification. AB - We present a rapid method for the identification of viruses using microfluidic chip gel electrophoresis (CGE) of high-copy number proteins to generate unique protein profiles. Viral proteins are solubilized by heating at 95 degrees C in borate buffer containing detergent (5 min), then labeled with fluorescamine dye (10 s), and analyzed using the microChemLab CGE system (5 min). Analyses of closely related T2 and T4 bacteriophage demonstrate sufficient assay sensitivity and peak resolution to distinguish the two phage. CGE analyses of four additional viruses--MS2 bacteriophage, Epstein-Barr, respiratory syncytial, and vaccinia viruses--demonstrate reproducible and visually distinct protein profiles. To evaluate the suitability of the method for unique identification of viruses, we employed a Bayesian classification approach. Using a subset of 126 replicate electropherograms of the six viruses and phage for training purposes, successful classification with non-training data was 66/69 or 95% with no false positives. The classification method is based on a single attribute (elution time), although other attributes such as peak width, peak amplitude, or peak shape could be incorporated and may improve performance further. The encouraging results suggest a rapid and simple way to identify viruses without requiring specialty reagents such as PCR probes and antibodies. PMID- 19551976 TI - Gold nanoparticle-based colorimetric and "turn-on" fluorescent probe for mercury(II) ions in aqueous solution. AB - An approach for visual and fluorescent sensing of Hg2+ in aqueous solution is presented. This method is based on the Hg(2+)-induced conformational change of a thymine (T)-rich single-stranded DNA (ssDNA) and the difference in electrostatic affinity between ssDNA and double-stranded (dsDNA) with gold nanoparticles. The dye-tagged ssDNA containing T-T mismatched sequences was chosen as Hg2+ acceptor. At high ionic strength, introduction of the ssDNA to a colloidal solution of the aggregates of gold nanoparticles results in color change, from blue-gray to red of the solution, and the fluorescence quenching of the dye. Binding of Hg2+ with the ssDNA forms the double-stranded structure. This formation of dsDNA reduces the capability to stabilize bare nanoparticles against salt-induced aggregation, remaining a blue-gray in the color of the solution, but fluorescence signal enhancement compared with that without Hg2+. With the optimum conditions described, the system exhibits a dynamic response range for Hg2+ from 9.6 x 10( 8) to 6.4 x 10(-6) M with a detection limit of 4.0 x 10(-8) M. Both the color and fluorescence changes of the system are extremely specific for Hg2+ even in the presence of high concentrations of other heavy and transition metal ions, which meet the selective requirements for biomedical and environmental application. The combined data from transmission electron microscopy, fluorescence anisotropy measurements, and dialysis experiments indicate that both the color and the fluorescence emission changes of the DNA-functioned gold nanoparticles generated by Hg2+ are the results of the metal-induced formation of dsDNA and subsequent formation of nanoparticle aggregates. PMID- 19551978 TI - Characterization of individual nano-objects by secondary ion mass spectrometry. AB - We present secondary ion mass spectrometry (SIMS) data obtained from the bombardment of a novel nanomaterial with a suite of projectiles: Au1+, Au3+, Au9+, and Au400(4+). These are the first experiments where free-standing nano objects were bombarded with kiloelectronvolt projectiles of atomic to nanoparticle size (Au400(4+)). The objects are aluminum monohydrate nanowhiskers, identified as crystalline boehmite (AlOOH) using X-ray diffraction. The nanoalumina is bonded to a microglass fiber that serves as a scaffold. The largest projectile, Au400(4+), has a diameter of approximately 2 nm, comparable to the nominal diameter of the nanowhiskers. There are notable differences in secondary ion (SI) response from sample volumes too small for full projectile energy deposition. The whisker spectra are dominated by small clusters--the most abundant species being AlO- and AlO2-. Bulk samples have larger yields for AlO2- than AlO-, whereas this trend is reversed in the whisker samples. Bulk samples give similar abundances of large SI cluster families [(Al2O3)(n)AlO2]- and [(Al2O3)(n)OH]-, whereas the whisker samples give an order of magnitude lower yield of these SIs. Given the nature of our experiments, i.e., the event-by-event bombardment/detection mode, we are uniquely able to obtain information from SIs emitted from single-projectile impacts. As such, effective yields were calculated in order to determine quantitative differences between the nano-objects and bulk samples. PMID- 19551977 TI - Enhanced digestion efficiency, peptide ionization efficiency, and sequence resolution for protein hydrogen/deuterium exchange monitored by Fourier transform ion cyclotron resonance mass spectrometry. AB - Solution-phase hydrogen/deuterium exchange (HDX) monitored by high-resolution Fourier transform ion cyclotron resonance (FTICR) mass spectrometry offers a rapid method to study protein conformations and protein-protein interactions. Pepsin is usually used to digest proteins in HDX and is known for lack of cleavage specificity. To improve digestion efficiency and specificity, we have optimized digestion conditions and cleavage preferences for pepsin and protease type XIII from Aspergillus saitoi. A dilution series of the proteases was used to determine the digestion efficiency for several test proteins. Protease type XIII prefers to cleave on the C-terminal end of basic amino acids and produced the highest number of fragments and the best sequence coverage compared to pepsin or protease type XVIII from Rhizhopus. Furthermore, protease type XIII exhibited much less self-digestion than pepsin and thus is superior for HDX experiments. Many highly overlapped segments from protease type XIII and pepsin digestion, combined with high-resolution FTICR mass spectrometry, provide high sequence resolution (to as few as one or two amino acids) for the assignment of amide hydrogen exchange rate. Our H/D exchange results correlate well with the secondary and tertiary structure of myoglobin. Such assignments of highly overlapped fragments promise to greatly enhance the accuracy and sequence resolution for determining conformational differences resulting from ligand binding or protein-protein interactions. PMID- 19551979 TI - Impedance spectroscopy of bacterial membranes: coenzyme-Q diffusion in a finite diffusion layer. AB - The inner membrane of Escherichia coli, overexpressing an ubiquinol oxidase, cytochrome bo3 (cbo3), was "tethered" in a planar configuration to a gold electrode. Electron transfer to cbo3 was achieved via native ubiquinol-8 or added ubiquinol-10, and impedance spectroscopy was used to characterize the diffusion properties of the ubiquinol/ubiquinone in the tethered membrane system. Spectra were obtained at varying direct current (DC) potentials covering the potential window in which the voltammetric catalytic wave of cbo3 is visible. These spectra were compared to those obtained after addition of a potent inhibitor of cbo3, cyanide, and the difference in impedance was analyzed using a derived equivalent circuit, which is similar to that of open finite-length diffusion (OFLD) or the finite Warburg circuit, but with the boundary conditions modified to account for the fact that ubiquinol reoxidation is limited by enzyme activity. Analysis of the impedance spectra of the tethered membrane system gave kinetic parameters that are consistent with values obtained using cyclic voltammetry. Importantly, the diffusion rate of ubiquinone (10(-13)-10(-12) cm2/s) was found to be orders of magnitude lower than accepted values for lateral diffusion (10(-8)-10(-7) cm2/ s). It is hypothesized that this result represent perpendicular diffusion of quinone across the membrane, corresponding to a "flip" time between 0.05 and 1 s. PMID- 19551980 TI - Low-temperature plasma probe for ambient desorption ionization. AB - A low-temperature plasma (LTP) probe has been developed for ambient desorption ionization. An ac electric field is used to induce a dielectric barrier discharge through use of a specially designed electrode configuration. The low-temperature plasma is extracted from the probe where it interacts directly with the sample being analyzed, desorbing and ionizing surface molecules in the ambient environment. This allows experiments to be performed without damage to the sample or underlying substrate and, in the case of biological analysis on skin surfaces, without electrical shock or perceptible heating. Positive or negative ions are produced from a wide range of chemical compounds in the pure stateand as mixtures in the gaseous, solution, or condensed phases, using He, Ar, N2, or ambient air as the discharge gas. Limited fragmentation occurs, although it is greater in the cases of the molecular than the atomic discharge gases. The effectiveness of the LTP probe has been demonstrated by recording characteristic mass spectra and tandem mass spectra of samples containing hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and 2,4,6-trinitrotoluene (TNT) from poly(tetrafluoroethylene) (PTFE) surfaces where limits of detection are as low as 5 pg. Other performance characteristics, when using a commercial ion trap mass spectrometer, include 3-4 orders of magnitude linear dynamic range in favorable cases. Demonstration applications include direct analysis of cocaine from human skin, determination of active ingredients directly in drug tablets, and analysis of toxic and therapeutic compounds in complex biological samples. Ionization of chemicals directly from bulk aqueous solution has been demonstrated, where limits of detection are as low as 1 ppb. Large surface area sampling and control of fragmentation by a simple adjustment of the electrode configuration during operation are other demonstrated characteristics of the method. PMID- 19551982 TI - Paper-based microfluidic devices by plasma treatment. AB - Paper-based microfluidic patterns have been demonstrated in recent literature to have a significant potential in developing low-cost analytical devices for telemedicine and general health monitoring. This study reports a new method for making microfluidic patterns on a paper surface using plasma treatment. Paper was first hydrophobized and then treated using plasma in conjunction with a mask. This formed well defined hydrophilic channels on the paper. Paper-based microfluidic systems produced in this way retained the flexibility of paper and a variety of patterns could be formed. A major advantage of this system is that simple functional elements such as switches and filters can be built into the patterns. Examples of these elements are given in this study. PMID- 19551981 TI - Novel three-dimensional MALDI plate for interfacing high-capacity LC separations with MALDI-TOF. AB - Novel matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) sample plates employing collimated-hole structures have been developed that allow capture and concentration of samples while simultaneously acting as a sink for carrier solvents. These plates were designed to provide an efficient interface between higher-capacity liquid chromatography (LC) separations and MALDI-TOF mass spectrometry (MS). LC-MALDI using conventional plates can accommodate the low flow (< 1 microL/min) separation protocols typically used in on-line LC-MS methods, and can also be used with higher flow rate, larger columns, but are ultimately limited by the capacity of the two-dimensional surface onto which the sample is deposited. Typically, about 1 microL of chromatographic effluent plus 1 microL of matrix solution can be deposited and dried on a ca. 3 mm diameter spot. Deposition rates (spot dwell time) are determined by the chromatographic resolution and the flow rate. To overcome this limitation, a new three dimensional MALDI sample plate has been developed using collimated-hole structures (CHS) with monolithic chromatography media filling the holes in the collimated-hole structures. These new plates retain all of the required features of conventional sample plates, commonly formed from stainless steel, but provide additional capacity for capturing and concentrating samples. Results are presented from reversed-phase separation of peptides on a 1 mm i.d. column operating at flow rate of 50 microL/min. Typically, 10 microL of effluent can be collected on a single spot, and sample and matrix dried on a 1 mm diameter spot, to yield about 30-fold enrichment of sample concentration in matrix crystals on the surface compared to the conventional plate. Sample loadings ranging from 1 fmol to 10 pmol/spot were investigated. PMID- 19551983 TI - Single-cell chemical lysis method for analyses of intracellular molecules using an array of picoliter-scale microwells. AB - Analyzing the intracellular contents and enzymatic activities of single cells is important for studying the physiological and pathological activities at the cellular level. For this purpose, we developed a simple single-cell lysis method by using a dense array of microwells of 10-30-pL volume fabricated by poly(dimethylsiloxane) (PDMS) and a commercially available cell lysis reagent. To demonstrate the performance of this single-cell lysis method, we carried out two different assays at the single-cell level: detection of proteins by antibody conjugated microbeads and measurement of protease activity by fluorescent substrates. The results indicated that this method readily enabled us to monitor protein levels and enzymatic activities in a single cell. Because this method required only an array of PDMS microwells and a fluorescence microscope, the simplicity of this platform opens a way to explore the biochemical characteristics of single cells even by those who are not familiar with microfluidic technology. PMID- 19551984 TI - Impedance biosensor for peanut protein Ara h 1. AB - A reagentless electrochemical impedance biosensor for detection of peanut protein Ara h 1, one of the allergenic proteins found in peanuts, has been demonstrated using an Au substrate onto which an antibody film has been immobilized. Following initial stabilization of the self-assembled monolayer (SAM) through which the antibody is immobilized, the biosensor substrate exhibits stable impedance spectra at different stages of substrate preparation. By fitting the impedance spectra to a Randles equivalent circuit, one can demonstrate that the charge transfer resistance (R(ct)) increases and the differential capacitance (Cd) decreases with increasing concentration of Ara h 1, although R(ct) exhibits greater sensitivity. The detection limit of this reagentless biosensor is estimated to be less than 0.3 nM. Assuming a Langmuir adsorption isotherm, the dissociation constant of the peanut protein Ara h 1 and its antibody can be calculated as 0.52 nM from the variation in differential capacitance with Ara h 1 concentration. PMID- 19551985 TI - Design and sensing properties of an integrated optical gas sensor based on a multilayer structure. AB - In this paper, a new multilayer integrated optical sensor (MIOS) for ammonia detection at room temperature is proposed and characterized. The sensor is integrated on a single-mode TE0-TM0 planar polymer waveguide and based on polyaniline (PANI) sensitive material. A polymethyl methacrylate (PMMA) passive layer is deposited between the waveguide core and PANI sensitive layer in order to decrease optical losses induced by evanescent wave/sensitive material coupling. The design of this new sensor is discussed. Moreover, in order to investigate the feasibility of this sensor, the sensing properties to ammonia at room temperature are studied. A significant change is observed in the guided light output power after the sensor is exposed to ammonia gas, due to PANI absorption coefficient variation. This new ammonia sensor shows fast response and recovery times, good reversibility and repeatability. The metrological parameters (sensitivity, response time and recovery time) of the sensor are strongly influenced by the interaction length (length of sensing region) and the PANI forms (doped and dedoped). The sensor has a logarithmic linear optical response within the ammonia concentration range between 92 to 4618 ppm. These experimental results demonstrate that the MIOS structure presents a potential innovation to elaborate integrated optical sensor based on non transparent (opaque) sensitive material. PMID- 19551987 TI - Effects of binding of metals to the hydrogel and filter membrane on the accuracy of the diffusive gradients in thin films technique. AB - The effect of metal binding to the diffusion layer of diffusive gradients in thin films (DGT) on measurements of time-integrated concentrations (CDGT) was studied experimentally and using a numerical model. Cationic divalent Cu and Cu-fulvic complexes bind to sites in the filter membrane and polyacrylamide (APA) gel, together comprising the diffusion layer of DGT. Less Cu was found in Chelex gels retrieved immediately after DGT deployment than in Chelex gels retrieved after storage of deployed DGTs. The difference was attributed to postdeployment transport of Cu from the diffusion layer to the Chelex gel. Binding of Cu in the diffusion layer delays the formation of a steady concentration gradient between solution and Chelex gel and can, depending on diffusion layer thickness, speciation, and concentration of metal, significantly bias the measurement of CDGT for short deployment times. However, C(DGT) will converge with the concentration in solution provided that the deployment time is sufficiently long (for a 1.0 mm diffusion layer, a diffusion coefficient of 5.5 x 10(-6) cm2 s(-1) and a concentration of Cu in solution of 0.1 nM, the difference is less than 11% and 5% for deployment times longer than 32 and 64 h, respectively). Minimal bias is encountered when measuring Cu contaminated waters (> or = 100 nM) using DGTs with thinner (< or = 0.5 mm) diffusion layer thicknesses. Diffusion layer binding of cationic Cd in the presence of cationic Cu was insignificant, and C(DGT) of Cd was found to converge with the concentration in solution after deployment times of 4 h and less. These findings have implications for the maximum temporal resolution that can be achieved with DGT. PMID- 19551986 TI - Software tool for the structural determination of glycosaminoglycans by mass spectrometry. AB - Structural elucidation of glycosaminoglycans (GAGs) is one of the major challenges in biochemical analysis. This is mainly because of the diversity of GAG sulfation and N-acetylation patterns and variations in uronate isomers. ESI MS and recently MALDI-MS methodologies are important strategies for investigating the molecular structure of GAGs. However, the interpretation of MS data produced by these strategies must take into account a large number of variables (including the number of monosaccharide residues, acetylations, sulfate groups, multiple charges, and exchanges between different cations). We have developed a bioinformatics tool to assist this complex interpretation task. The software is based on GlycoWorkbench, a tool for semiautomatic interpretation of glycan MS data. The tool generates the sugar backbones in all their variants (GAG family, composition, acetylation positions, and number of sulfates) and automatically matches them with the selected MS peaks. The backbones corresponding to a given peak are validated against the selected MS/MS peaks by generating all possible fragmentations. Native chondroitin sulfate and heparin oligosaccharides as well as chemically modified heparin oligomers have been successfully analyzed by MALDI and ESI-MS and MS/MS, and the results of the semiautomated annotation of these mass spectra are presented here. PMID- 19551988 TI - Detection of fluoroquinolone antibiotics in milk via a labeless immunoassay based upon an alternating current impedance protocol. AB - This paper describes the construction of a labeless immunosensor for the antibiotic ciprofloxacin in milk and its interrogation using an ac impedance protocol. Commercial screen-printed carbon electrodes were used as the basis for the sensor. Polyaniline was electrodeposited onto the sensors and then utilized to immobilize a biotinylated antibody for ciprofloxacin using classical avidin biotin interactions. Antibody loaded electrodes were exposed to solutions of antigen in milk and interrogated using an ac impedance protocol. The faradaic component of the impedance of the electrodes was found to increase with increasing concentration of antigen. Control samples containing a nonspecific IgG antibody were also studied but were found to display large nonspecific responses, probably due to the antibody binding some of the large number of components found in milk. Control sensors could, however, be fabricated using antibodies specific for species not found in milk. Calibration curves could be obtained by subtraction of the responses for specific and control antibody-based sensors, thereby eliminating the effects of nonspecific adsorption of antigen. Sensors exposed to ciprofloxacin in milk gave increases in impedance whereas ciprofloxacin in phosphate buffer led to decreases, indicating the possibility of developing sensors which can both detect and differentiate between free and chelated antigen. PMID- 19551989 TI - Biosensor-based screening method for the detection of aflatoxins B1-G1. AB - Aflatoxins are extremely toxic metabolites from Aspergillus species that can adulterate a wide range of human foodstuff. Herein, we propose a novel assay designed as an analytical test for aflatoxin B1 and G1 (AFB1 and AFG1, respectively) that could represent an alternative screening technique for this class of mycotoxins. The approach for the determination of these toxins is based on surface plasmon resonance using neutrophil porcine elastase as a "bait" for these aflatoxins. The selection and optimization of the analytical procedure involved a preliminary investigation on the type of inhibition by AFB1: the level of the protease inhibition exerted by AFB1 depended upon the incubation time and the concentration of the binding partners, showing the competitiveness and the reversibility of the inhibition. A posteriori, the nature of the interaction granted a rapid analysis, a single detection test requiring only a few minutes. For the development of the assay, the experimental conditions were evaluated and optimized with both calibration solution and aflatoxin-spiked samples. To apply this method to aflatoxin-contaminated maize, a rapid solid-phase extraction treatment was developed. The proposed assay for AFB1 and AFG1 was validated by comparison with both a chromatographic reference method and a standard enzyme linked immunosorbent assay procedure. This enzyme-based biosensor represents a new approach for the detection of aflatoxins based on the reversible interaction between a blocked macromolecule and a soluble ligand, having the major advantages in the relative rapidity, the reusability of the capturing surface, and low cost per single test. PMID- 19551990 TI - Electrochemical immunosensor for detection of celiac disease toxic gliadin in foodstuff. AB - Celiac disease is a gluten-sensitive enteropathy that affects as much as 1% of the population. Patients with celiac disease should maintain a lifelong gluten free diet, in order to avoid serious complications and consequences. It is essential to have methods of analysis to reliably control the contents of gluten free foods, and there is a definitive need for an assay that is easy to use, and can be used on site, to facilitate the rapid testing of incoming raw materials or monitoring for gluten contamination, by industries generating gluten-free foods. Here, we report on the development of an electrochemical immunosensor exploiting an antibody raised against the putative immunodominant celiac disease epitope, for the measurement of gliadin content and potential celiac toxicity of a foodstuff. To develop the gliadin immunosensor, we explored the use of two surface chemistries, based on the use of dithiols, 22-(3,5-bis((6 mercaptohexyl)oxy)phenyl)-3,6,9,12,15,18,21-heptaoxadocosanoic acid (1) and 1,2 dithiolane-3-pentanoic acid (thioctic acid) (2), for anchoring of the capture antibody. The different surface chemistries were evaluated in terms of time required for formation of self-assembled monolayers, stability, susceptibility to nonspecific binding, reproducibility, and sensitivity. The thioctic acid self assembled monolayer took more than 100 h to attain a stable surface and rapidly destabilized following functionalization with capture antibody, while the heptaoxadocosanoic acid surface rapidlyformed (less than 3 h) and was stable for at least 5 days, stored at room temperature, following antibody immobilization. Both surface chemistries gave rise to highly sensitive immunosensors, with detection limits of 5.5 and 11.6 ng/mL being obtained for 1 and 2, respectively, with nonspecific binding of just 2.7% of the specific signal attained. The immunosensors were extremely reproducible, with RSD of 5.2 and 6.75% obtained for 1 and 2 (n = 5, 30 ng/mL), respectively. Finally, the immunosensor was applied to the analysis of commercial gluten-free and gluten-containing raw and processed foodstuffs, and excellent correlation achieved when its performance compared to that of an ELISA. PMID- 19551991 TI - Ionic reagent for controlling the gas-phase fragmentation reactions of cross linked peptides. AB - Chemical cross-linking combined with proteolytic digestion and mass spectrometry (MS) is a promising approach to provide inter- and intramolecular distance constraints for the structural characterization of protein topologies and functional multiprotein complexes. Despite the relative straightforwardness of these methodologies, the identification and characterization of cross-linked proteins presents a significant analytical challenge, due to the complexity of the resultant peptide mixtures, as well as the array of inter-, intra-, or "dead end"-cross-linked peptides that may be generated from a single cross-linking experiment. To address these issues, we describe here the synthesis, characterization, and initial evaluation of a novel "fixed charge" sulfonium ion containing crosslinking reagent, S-methyl 5,5'-thiodipentanoylhydroxysuc cinimide. The peptide products obtained by reaction with this reagent are all shown to fragment exclusively via facile cleavage of the C-S bond directly adjacent to the fixed charge during CID-MS/MS, resulting in the formation of characteristic product ions that enable the presence and type (i.e., inter, intra, or dead-end) of the cross-linked products to be readily determined, independently of the "proton mobility" of the precursor ion. Subsequent isolation and dissociation of these products by MS3 provides additional structural information required for identification of the peptide sequences involved in the cross-linking reactions, as well as for characterization of the specific site(s) at which cross-linking has occurred. The specificity of these gas-phase fragmentation reactions, as well as the solubility and stability of the cross linking reagent under aqueous conditions, suggests that this strategy holds great promise for use in future studies aimed at the structural analysis of large proteins or multiprotein assemblies. PMID- 19551993 TI - Imaging localized astrocyte ATP release with firefly luciferase beads attached to the cell surface. AB - Extracellular adenosine triphosphate (ATP) functions as a signaling molecule in many cell regulation processes. The traditional firefly luciferase assays measure the ATP release as a signal increase with time using a luminometer. Recently, advanced cell imaging techniques using charge-coupled device (CCD) cameras have enabled two-dimensional (2D) high-resolution detection providing both spatial and temporal information. Real-time imaging of ATP release from astrocyte cells has been reported. However, the observed chemiluminescence propagation wave reflects both ATP release and diffusion in the extracellular bulk solution. The dynamic ATP efflux at the cell surface could not be accurately measured. Hence, we constructed biotinylated fused firefly luciferase proteins, immobilized the proteins on 1 microm beads, and attached the beads to the cell surface to detect ATP release from mechanically stimulated astrocyte cells. This novel detection method enables us to monitor the actual ATP concentration at the surface of single live cells. The localized ATP release was found to be prominent but lasted only <20 s, which is very different from the results obtained by free firefly luciferase detection. PMID- 19551992 TI - Multiple isotopic labels for quantitative mass spectrometry. AB - Quantitative mass spectrometry is often performed using isotopically labeled samples. Although the 4-trimethylammoniumbutyryl (TMAB) labels have many advantages over other isotopic tags, only two forms have previously been synthesized (i.e., a heavy form containing nine deuteriums and a light form without deuterium). In the present report, two additional forms containing three and six deuteriums have been synthesized and tested. These additional isotopic tags perform identically to the previously reported tags; peptides labeled with the new TMAB reagents coelute from reversed-phase HPLC columns with peptides labeled with the lighter and heavier TMAB reagents. Altogether, these four tags allow for multivariate analysis in a single liquid chromatography/mass spectrometry analysis, with each isotopically tagged peptide differing in mass by 3 Da per tag incorporated. The synthetic scheme is described in simple terms so that a biochemist without specific training in organic chemistry can perform the synthesis. The interpretation of tandem mass spectrometry data for the TMAB labeled peptides is also described in more detail. The additional TMAB isotopic reagents described here, together with the additional description of the synthesis and analysis, should allow these labels to be more widely used for proteomics and peptidomics analyses. PMID- 19551994 TI - Nitroavidin as a ligand for the surface capture and release of biotinylated proteins. AB - Discussed here is the preparation, detailed purification, and evaluation of nitroavidin as a ligand for surface capture and release of biotinylated proteins. Avidin from chicken egg white was nitrated using dilute tetranitromethane solutions. UV-vis spectroscopy was used to show decreased binding of the biotin analogue, 2-(4'-hydroxyazobenzene)benzoic acid, HABA, to nitroavidin compared to binding of HABA to native avidin. From enzyme-linked immunosorbent assay (ELISA) based assays of the modified avidin, it was found that there are approximately three tyrosine residues converted to nitrotyrosine out of the total four tyrosine residues in the protein tetramer. For the first time, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to demonstrate the point of nitration in nitroavidin as that of the tyrosine associated with the binding of biotin (Y33). From surface plasmon resonance spectroscopy (SPR) experiments, it is shown that biotin has less binding propensity to immobilized nitroavidin (K(D) = 4.4 +/- 1.9 x 10(-6) M) than immobilized avidin (K(D) < or = 10(-11) M). Importantly, the use of pH 10 carbonate buffer as eluent resulted in facile release of bound biotin from the nitroavidin-functionalized surfaces, allowing for readily regenerated biotin capture surfaces (reversible binding surfaces). These outcomes are important for the development of protein concentration methods directed at isolation of select proteins from a large population using gentle target protein isolation/release conditions. PMID- 19551995 TI - Construction of time-lapse scanning electrochemical microscopy with temperature control and its application to evaluate the preservation effects of antifreeze proteins on living cells. AB - Antifreeze proteins (AFPs) can protect cells from hypothermic damage; however, their mechanism of action remains unclear. Scanning electrochemical microscopy (SECM) can evaluate the size and activities of cells, although long-term continuous monitoring has been unsuccessful. We constructed a novel, fully automated, time-lapse SECM system and investigated the cell preservation effect of AFPs by analyzing single cellular topography at low temperatures. From the SECM measurements, mammalian cells (HepG2), treated in Euro-Collins (EC) solution at 4 degrees C, began to swell at 8 h and then immediately ruptured. In AFP containing EC solution, the cellular size did not change until 16 h and then gradually increased and finally ruptured. In addition, the cellular height at rupture point significantly increased in the presence of AFPs. These results suggest that AFPs stabilize the cellular membrane and protect cells from hypothermic damage. This SECM system allowed us to observe the single cellular response to hypothermia by long-term automatic scanning and will be applicable for analysis to other cellular activities and topographies. PMID- 19551997 TI - Microwave-induced combustion coupled to flame furnace atomic absorption spectrometry for determination of cadmium and lead in botanical samples. AB - A procedure based on microwave-induced combustion coupled to flame furnace (FF) atomic absorption spectrometry (FF-AAS) was used for analysis of solid samples. Botanical samples were prepared as pellets and introduced into a quartz holder device. This device was fitted to a glass chamber that was used for the combustion step. The complete device was coupled to the flame furnace by using poly(tetrafluoroethylene) and quartz tubes. The glass chamber was placed inside a microwave oven in a position previously set to receive the higher power of microwave radiation. Ignition was performed by microwave radiation using a small piece of paper wetted with NH4NO3 solution. An oxygen flow was used to assist the sample combustion and also to transport the combustion products up to the heated FF positioned above an air/ acetylene burner. Flame furnace temperature, oxygen flow rate, flame stoichiometry, and sample mass range were evaluated. Cadmium and lead were determined in botanical samples as examples to demonstrate the potential of the proposed procedure for trace analysis. Sample masses up to 60 mg could be used, allowing a limit of detection as low as 0.003 and 0.24 microg g( 1) for Cd and Pb, respectively. Integrated absorbance was used with an integration time of 30 s. Background signals were always low, and relative standard deviation (n = 5) was below 9% for Cd and 11% for Pb. The throughput was 20 determinations/h, including the weighing step. Accuracy was between 94 and 105%, and calibration was performed using standard solutions. The combustion device could be easily adapted to conventional atomic absorption spectrometers. PMID- 19551996 TI - Dequenching of Cu(I)-bathocuproine disulfonate complexes for high-performance liquid chromatographic determination of phytochelatins, heavy-metal-binding peptides produced by the primitive red alga Cyanidioschyzon merolae. AB - A novel method has been devised for the determination of phytochelatins (PCs), heavy-metal-tolerant peptides produced by higher plants and algae. The method is based on the facts that fluorescence of bathocuproine disulfonate (BCS) is quenched by Cu(I) ions as a result of Cu(I)-BCS complex formation and that PCs compete with BCS for Cu(I). Detection of PCs via recovered fluorescence of BCS using the Cu(I)-BCS complex as a postcolumn reagent, following separation of peptides on an octyldecylsilane column, demonstrated a highly sensitive method for determination of PCs. PCs in the primitive red alga, Cyanidioschyzon merolae, grown in the presence or absence of added Cd(II) were successfully determined by this protocol. Unlike other methods for the determination of PCs, which rely on the SH groups in the peptides, the proposed method is unique in that detection is based on the chemical nature of PCs, which favors the formation of complexes with Cu(I). In this context, the new method yields chromatograms based on the strength of binding Cu(I) ions. PMID- 19551998 TI - Diabetes and the downturn. Getting care during tough times. PMID- 19551999 TI - Fast-food cooking at home. Fresh fish. PMID- 19552001 TI - Diabetes quiz. How much do you know about indigestion? PMID- 19552000 TI - Managing chronic pain. PMID- 19552002 TI - Keeping active with diabetes and arthritis. PMID- 19552003 TI - Barley the Italian way. PMID- 19552004 TI - Supermarket smarts. Canned chili. PMID- 19552005 TI - Evaluating construct validity of the Slosson Visual-Motor Performance Test using the Rasch Measurement Model. AB - The aim of this study was to evaluate the construct validity of the Slosson Visual-Motor Performance Test by applying the Rasch Measurement Model to evaluate the test's scalability, dimensionality, differential item functioning based on sex, and hierarchical ordering. Participants were 400 children ages 5 to 12 years, recruited from six schools in Melbourne, Victoria, Australia. The Slosson Visual-Motor Performance Test requires a child to copy 14 different geometric designs three times each for a total 42 scale items. Children completed the test under the supervision of an occupational therapist. Overall, 13 of 42 of the test items exhibited poor measurement properties. As nearly one-third of the scale items were problematic, the Slosson Visual-Motor Performance Test in its current form is not recommended for clinical use. PMID- 19552006 TI - [Abstracts of the Portuguese Pneumology Society XXII Pneumology Congress and the IV Luso-Brasileiro Pneumology Congress]. PMID- 19552007 TI - 1-Cyano-4-dimethylamino pyridinium tetrafluoroborate as a cyanylating agent for the covalent attachment of ligand to polysaccharide resins. PMID- 19552008 TI - Limitations of N-hydroxysuccinimide esters in affinity chromatography and protein immobilization. AB - The carbodiimide-mediated reaction of N-hydroxysuccinimide with carboxyl groups immobilized to hydroxyl-containing polymers (such as Sepharose or Trisacryl) leads to an undesirable side reaction in high yields. The product of this reaction interferes with the application of such columns for further affinity based purification. In addition to the desired N-hydroxysuccinimide ester, a bis(N-hydroxysuccinimide)derivative of beta-alanine [namely, N- [(succinimidooxy)carbonyl]-beta-alanine N-hydroxysuccinimide ester] is probably produced that reacts subsequently with the hydroxyl group of the polymer via ester and carbamate bonds. These beta-alanine derivatives are formed upon interaction of dicyclohexylcarbodiimide with 3 equiv of N-hydroxysuccinimide followed by a Lossen rearrangement. The amount of beta-alanine thus coupled is very high compared to the number of carboxyl groups present on the resin. The beta-alanine bound through the ester bond comprises about 90% of the beta-alanine bound. Alkaline treatment of the ester-bonded beta-alanine-containing polymers (prior to coupling of amino-containing ligands) causes a rearrangement yielding beta-alanine with a free carboxyl group coupled through a stable carbamate linkage. After coupling of amino-containing ligands, the above-described rearrangement cannot occur, and the beta-alanine-linked ligand leaks from the polymer via hydrolysis of the ester bond. The newly formed carboxyl groups (derived from the rearrangement) can be used to prepare active esters. In view of the above, we developed methods for the preparation of nitrophenyl esters as well as N-hydroxysuccinimide esters free of unstable beta-alanine derivatives on polymers containing hydroxyl groups. Upon coupling with amino-containing ligands, these esters yield resins bearing chemically stable bonds. PMID- 19552009 TI - [Ovarian hyperstimulation syndrome]. AB - Experience with ovulation induction treatment and understanding of the physiopathology of ovarian hyperstimulation syndrome (OHS), the risk factors and the clinical characteristics are of key importance for preventing and managing OHS. Light manifestations ofOHS are fairly common, occurring in up to a third of the cycles with superovulation induced by exogenous gonadotropins. A worsening of the symptoms of OHS can still normally be managed in out-patient form, but frequent monitoring and evaluation are essential. Serious disease resulting from OHS is much less common, but can involve risk to life. Hospitalisation might be necessary in some cases. PMID- 19552010 TI - Ciencia & Saude Coletiva. Editorial. PMID- 19552011 TI - Exercise for management and treatment of Parkinson disease. PMID- 19552012 TI - [ERCC1 and RRM1 genes in lung cancer]. AB - In lung cancer, expressive survival has not yet been achieved in non surgical stages. Non-small cell lung cancer (NSCLC) patients are treated with platinum and other drugs. To choose these agents we can actual ly define predictive biomarkers to preview therapeutic response. A literature revision was done in order to define the role of ERCC1 e RRM1 genes in the response to chemotherapy based in platinum and gemcitabine respectively. The expression of these genes is faced as a predictive marker to the chemotherapy response in patients with adenocarcinomas and squamous cell carcinomas, providing a personalized therapy. Published data supports this behaviour and is useful to individualize therapy accordingly to individual levels of ERCC1 which are modified by genetic mutations. Polymorphisms in codons 118 C/T and C8092A, seem to influence the carcinogenesis, cytostatic resistance, survival and even the prognosis. Clinical and laboratorial trials showed that high expression of RRM1 gene in NSCLC has impact in the tumoral phenotype. Patients having done surgical resection and presenting high expression of RRM1 have better survival than those with lower expression. However, patients with advanced NSCLC and treated with chemotherapy with gemcitabine and cisplatin appear to have a poor outcome if the tumor express elevated levels of RRM1 gene. PMID- 19552013 TI - [Postoperative pain management: guidelines, organization and techniques]. PMID- 19552015 TI - [ABO hemolytic disease and developing of significant hyperbilirubinemia in term newborns: early predictive factors]. PMID- 19552016 TI - Guest editor's introduction: special section on shape, solid, and physical modeling. PMID- 19552017 TI - [Efficiency of famotidin in prophylaxis of NSAIDs-induced gastropathy: result of multicenter research ZASLON-1 (protection of gastric mucosa from non-steroidal anti-inflammatory drugs]. AB - This article presents results of the first Russian randomized study of NSAID gastropaty prevention at patients receiving NSAIDS. Results of clinical and endoscopic monitoring of two groups of patients: 1st - 108 patients with AA or RA treated with diclofenac and famotidin and 2nd--116 patients with AA or RA receiving diclofenac treatment only. It was shown that combined use of diclofenac and famotidin reduces to 2.6 times frequency and significantly reduces severity of painful dyspepsia symptoms and antipain manifestations reduces more than twice frequency of erosive-ulcerous lesions of gastric mucosa and duodenum. PMID- 19552018 TI - [Present status of clinical and fundamental threpsology]. AB - A good nutrition status is one of the impotant prerequisites of good health. Malnutrition in hospital leads to increased complication, morbidity and mortality rates. Whenever the gastrointestinal tract cannot fulfil its function or must be rested (during intestinal desease), parenteral administration of nutrients (or simultative parenteral and enteral alimitation) is necessary. Nowday perspectives of nutrition support in patients whith intestinal deseases, methods of easily diagnosed of malnutrition are the problems of this article. PMID- 19552019 TI - [The state of the gastrointestinal tract and glucose metabolism as limiting factors of parenteral-enteral nutrition in patients with intracranial hemorrhages]. AB - A part of patients' arterial hyperglycemia is not accompanied by increases in the content of glucose in brain substance, and aggressive correction of hyperglycemia can lead to reduction in concentration of glucose expressed in "affected", and conditionally "intact" brain substance. At choosing a mixed strategy of parenteral-enteral correction should take into account not only condition of the gastrointestinal tract but especially absorption of glucose (the level of cellular homeostasis). PMID- 19552021 TI - [Nutritional support and efficient gastroduodenal endoscopy in the treatment of acute pancreatitis and different forms of pancreonecrosis]. AB - Nutrition support and molecular-genetic markers of the immune system at patients with complicated forms of gastric pathology. In this study presented results of surgery clinic of and oncology faculty SGMU to improve treatment patients with acute pancreatitis, and various forms of pancreonecrozis, through a balanced Threpsology support and operational gastrointestinal endoscopy. PMID- 19552020 TI - [Effectiveness of nutritional support for the liver-cell deficiency in the liver cirrhosis]. AB - THE AIM: To study the efficiency of enteral nutritional support and its impact on clinical-laboratorial markers of hepatocellular failure of the patients with Child-Pue's hepatic cirrhosis type B suffering from trafologic impairment. MATERIAL AND RESEARCH METHODS: 61 patients with Child-Pue's hepatic cirrhosis type B suffering from trafologic impairment took part in the randomized prospective study. The control group (31 patients) got the conventional treatment with lactulose, beta-blocker, spironolactone on the basis of 1.2 g/kg protein weight and 30 kcal/kg body weight per day. There were 30 patients in the second group (the principal one) and in addition to conventional treatment they got the multisubstrate nutritional cocktail of dietary fibers on the basis of 0.3 g/kg protein weight, 7.5 g fibers per day and 10 kcal/kg body weight during 30 days. To evaluate the efficiency of the treatment we analyzed the clinical presentation and the intensity of blood cholinesterase. The stage of hepatic encephalopathy was defined with the affinity numbers test of Retainer. The parameters under study were checked before the treatment, 30 days and 60 days later after the treatment began. RESULTS: It was found out during the prospective randomized study that the patients under nutritional support showed the positive clinical dynamics (asthenia and hepatic encephalopathy retrogression and the accurate increase of blood cholinesterase from 4365.4 +/- 1028.6 ME up to 5502.7 +/- 1142.6 ME (p < 0.05). CONCLUSION: Nutritional support of the patients with Child Pue's hepatic cirrhosis type B suffering from trafologic impairment is effective and has an accurate impact upon clinical-laboratorial markers of hepatocellular failure. PMID- 19552022 TI - [Bioactive systems based on nanostructured polymers for enteral nutrition]. AB - Nanostructured materials with various methods of coating have antimicrobial activity and can be used in creation of biologically active systems with a given medical-biological characteristics. Use of organic derivatives of fullerene S60 as a new class of biologically active compounds and their use is a new and promising direction in this area, providing a selective effect on the micro organisms of different species. Using probes with the biologically active nanostructured surfaces useful for the correction of pH-dependent states, such as the translocation of microflora in the intestine when postgastrectomical states may have to correct with GERD. Programmable output regulators of activity of intestinal microbiota can be viewed as a way of normalizing the digestive transport processes in the intestine. PMID- 19552023 TI - [Experimental and clinical evaluation of a modified pectin-containing drugs in the treatment of intestinal insufficiency in peritonitis]. AB - Experimental studies on 26 rats showed a stimulating effect of beet pectin at small intestine with paresis, mucosal repair, and bactericidal action on intestinal microflora. In comparison of activity intensity of small intestine solder at 1 day was 2.5-1.75 times less than 3 days and it is not reached. In the experimental group at 1 day solder activity was lower than the norm for the divisions in the 1.7-1.64--1.53-fold, and by 3 days, respectively, exceeded the norm of 0.8-2.4--2.25 times. We examined 150 patients aged 19 to 86 years with poured peritonitis primarily toxic (85.3%). III. The main causes of peritonitis were acute appendicitis (24.0%) and perforation of gastroduodenal ulcers (21.3%). For a comparative analysis of results of treatment were formed 3 patient groups using paired sample. Control group patients received conventional treatment. Enterosorption conducted through nazointestinalny probe known carbon sorbent UAO A and pektincontents products (SAPs) the dining beet sublimate materials. The results are compared. Reducing the time used to resolve paresis of intestine and nazointestinal intubation in 1,3 times, reducing the severity of the patient on a scale of SAPS is already at 2 hours improving immunity more rapid decline in toxicity of blood plasma and leukocyte index of intoxication, reduction of postoperative pneumonia in 12% and postoperative mortality from 14% to 6.6% showed the advantages enterosorption modified pectin contents drug. PMID- 19552024 TI - [Enteral nutrition in patients with ulcerative and postburn cicatrix strictures of the esophagus and stomach outcome region]. AB - Decompensated cicatrices stricture of upper alimentary canal is a complex disease clinically presenting a high mechanical blocking and leads to expressed abnormality of homeostasis, which requires its pathogenetic correction of urgency evidence. The greatest difficulty is correct protein-energy malnutrition and water-electrolyte metabolism. Prior to the imposition of stoma for feeding should begin immediately with standard parenteral nutrition solutions. In a subsequent it is nessesary to resort more physiologecal tube alimentasion. As with esophageal postambustion stricture electrical activity of the stomach inhibiting and in essentially remains small bowel function, preference should be given to ways of enteral threpsology support. This can be used as a balanced composition in breeding (primary breeding should be 1: 2) and special blends for intraintestinal alimentation (close chyme on line carrying the major components). In the case of postambustion struck of outlet termination stomach department when identified violations of the underlying functions of the digestive canal division, rational come to gentle tactics of enteral alimentation using mixtures, completely similar in composition to himus. At stricture janitor ulcer genesis appropriate tactics is enteral correction, similar to that used in the event of postambustion strictures of the zones when bowel function is largely preserved. PMID- 19552025 TI - [Hepato-renal syndrome and problem of protein-energy metabolism correction during urgent surgery of the abdominal organs]. AB - Early connecting of hardware detoxification methods allow more efficient use of amino acid preparations for action. The use of efferent technology provides a full correction of protein and energy metabolism in the immediate surgical patients with complicated diseases of the abdominal cavity. This will reduce bed day in patient and surgical mortality. PMID- 19552026 TI - [Barrett esophagus: modern diagnosis, drug therapy and reduce risk of cancer]. AB - This article describes ways of risk reducing of esophageal adenocarcinoma with the use of modern screening treatment methods of GERD and Barrett esophagus. PMID- 19552027 TI - [Evaluation of alpha-normiks (rifaximin) efficacy in the treatment of patients with diverticular disease associated with medium and severe intestinal dysbacteriosis]. AB - THE PURPOSE OF THE STUDY: to evaluate the efficacy of using rifaksimin drug within complex therapy of patients with the disease and diverticular intestinal dysbacteriosis. MATERIALS AND METHODS: 57 patients participated with a moderate exacerbation of the disease, they were divided equally on the composition of the group--two control and one under study. Patients in the control groups were composed of complex treatment of oral antibacterial drugs, or bacteriophage, the investigated group of patients--rifaksimin. We evaluated the dynamics of clinical symptoms, analysis of stool for dysbacteriosis and biochemical parameters. RESULTS: detected superior effectiveness of the Alpha-normiks drug in the treatment of patients with colon pathologies investigated in comparison with the use of oral antibacterial drugs, or bacteriophages. Proved the high safety of treatment with using of rifaksimin. CONCLUSIONS: The preparation of Alpha-normiks may be appointed with great success at patients with diverticular disease, and with intestinal dysbacteriosis. PMID- 19552028 TI - [Gastroesophageal reflux disease: pathogenetic basis of differentiated tactics of treatment]. AB - The article contains information about severe side effects of long-term acid suppression with non-select proton pump inhibitors (PPI) treatment of gastroesophageal reflux disease (GERD). As far as concern patients with heartborn without esophagitis (non-erosive GERD) using PPI doesn't correspond the pathogenesis and hardly has any advantages. Therapy with gastric acidity inhibitors increases risk of acute gastroenteritis and community-acquired pneumonia. Quality of life is rising and symptoms of gastro-esophageal reflux are getting away thanks to topical harmless treatment GERD without severe esophagitis Pepsan-R. PMID- 19552029 TI - [The effectiveness of spasmolytic therapy in chronic acalculous cholecystitis]. AB - The therapeutic efficacy of duspatalin was evaluated in 69 patients with chronic cholecystitis and dyskinesia of the biliary tract (BT). Supplementation of duspatalin to the combined therapy in the patients with chronic cholecystitis shown to exert a pronounced therapeutic effect. This caused positive changes in clinical symptoms and BT function and quality of life, diminished the lithogenic properties of bile. PMID- 19552030 TI - [The role and physiological basis of macro- and micronutrient metabolism in human nutrition]. AB - In this article presented original evidence on the beneficial effects of dyuspatalin (mebeverin) on the functional status of gallbladder and sphincter apparatus of biliary tract, as well as the lithogenic properties of bile at patients with chronic acalculous cholecystitis. PMID- 19552031 TI - [Mixtures for enternal nutrition in Russia (literature review)]. AB - The article provides a history of research in the field of enteral nutrition, history of the mixtures for tube alimentation. Also presented classification of the present mixtures for enteral introduction, as well as a characteristic of each of them. PMID- 19552032 TI - [Vegetarian diets in nutritional therapy]. PMID- 19552033 TI - [The state of the science of nutrition in the XXI century. From the theory of balanced nutrition to holistic theory]. AB - Holistic nutrition theory takes into account: Not only nutrition, but regulatory, sensory, landmark properties of food. Individual characteristics of people. Climatic, ethnic, social, occupational, religious, familial, aesthetic, ethical terms, and traditions; Conditions for use of new products including genetically modified. Food, as part of a biological culture, defining the correct behavior of man in the biosphere and noosphere. PMID- 19552034 TI - [Parenteral nutrition in the treatment of short bowel syndrome]. AB - Relevance of nutritive support problems at patients with SRL is shown in experiences of RSSC members named by B.V. Petrovski RAMS. This article demonstrates feasibility of artificial therapeutic nutrition and role of parenteral nutrition, clinical and social rehabilitation at patients with SRL. PMID- 19552035 TI - [Nutritional support in patients with acute pancreatitis]. AB - A causal therapy for acute pancreatitis is not available, so treatment has to focus on best supportive care. As nutrition is a cornerstone in the treatment of the disease, recent clinical research has focused on this question. The concept of enteral nutrition is aiming at the maintenance of a normal mucosal barrriere function which has been discovered to be important in mild and severe acute pancreatitis. This review offers an overview about current concepts of nutritional support and gives recommendations for clinical care as well. PMID- 19552036 TI - [Retrospective evaluation of the effectiveness of gastric enteral nutrition in intensive care unit]. AB - The duly hypocaloric intragastric nutrition is more portable and convenient to adapt in depressed consciousness patients of the intensive care unit (ICU). Daily caloric content of 16-19 kcal/kg does not allow to save the patient from significant protein losses that can be shown by serum albumin decreasing even in already restored peroral feeding. But there are no the certificated laboratory methods to indicate inadequate nutrition in due time. It is necessary to return to aprioristic taking the normocaloric feeding as the basis nutrition support of patients in ICU. PMID- 19552037 TI - Framework for action. PMID- 19552038 TI - Slowly rebuilding her life, the nurse jailed for a crime she did not commit. PMID- 19552039 TI - Does poor staffing affect job satisfaction and patient care? PMID- 19552040 TI - Flexible friends. AB - While the recession means that many families are economising, there is no shortage of demand for nurses with specialist skills to care for sick children at home. PMID- 19552041 TI - Caring in troubled times. AB - The risks to the quality of care in an uncertain economic crisis was the topic for debate at a fringe meeting at congress. PMID- 19552042 TI - Closer to home. AB - Having cancer chemotherapy can be a gruelling experience for patients. Taking a service closer to where they live can make all the difference. PMID- 19552043 TI - A cry for help. PMID- 19552044 TI - Improving services for people with alcohol-related problems. AB - From March 2007 to April 2008 the Royal College of Nursing supported a project to improve services for people with alcohol-related problems by demonstrating and publicising the effectiveness of nursing interventions. The project team included representation from England, Northern Ireland, Wales and Scotland. This article presents a framework that can be adapted and used by nurses to promote the development of specialist nursing services by health and social care commissioners and policy makers to improve care for people who misuse alcohol. PMID- 19552045 TI - Using ward-based simulation in cardiopulmonary training. AB - This article assesses the role of ward-based simulation in cardiopulmonary resuscitation training. Based on an initiative introduced at the Royal Hospital for Neuro-disability, London, it supports many of the positive claims made about the value of simulation in such contexts. The role of simulation was audited on the basis of feedback from staff, who indicated that the opportunity to practise their skills in a realistic setting increased confidence in their abilities to respond to a real-life event and promoted critical reflection. Further analysis indicated that, as a learning resource, simulation compares well with real-life experience, but is most effective when deployed alongside frequent updates based on more traditional teaching methods. PMID- 19552046 TI - A collaborative and progressive approach to basic life support. AB - This article discusses how emergency situations occur across a range of lay, community and hospital settings. Healthcare practitioners working in these environments need to be equipped to respond to these situations and be able to develop and expand on previously acquired skills and knowledge. They also need to be able to recognise and maximise other team members' contributions, especially in settings where there is a diverse interprofessional group. The approach outlined in this article is based on a model currently being used in a higher education institution. PMID- 19552047 TI - Hyperprolactinaemia. PMID- 19552048 TI - Key to student support. PMID- 19552049 TI - Keep it local. PMID- 19552050 TI - Unmask your talent. PMID- 19552051 TI - Can large animal experiments be totally replaced by alternatives? PMID- 19552052 TI - Neurobiology of cannabis addiction. AB - Cannabis has emerged as a common substance of abuse and dependence and the peculiarities associated with this widely available and used substance has triggered substantial research in this field. The earlier held concept of rather benign nature of this compound as a substance of abuse and dependence has changed as a result of the ongoing clinical and research findings. Cannabis has been found to have multiple physical and mental effects in human beings. But still a lot remains to be answered regarding the basis for the development of dependence on cannabis. However, the discovery of various cannabis receptors and their endogenous and synthetic ligands have added fuel to the ever growing interest in this substance. Various hypotheses have been postulated in this regard based on the findings of both the animal and human studies which serve as potential explanations to the observations. These findings have helped in the better understanding of the issue and have provided substrate for the clinical application. PMID- 19552053 TI - Protective effect of adenosine in diabetic neuropathic pain is mediated through adenosine A1-receptors. AB - Diabetic neuropathic pain is generally considered to be one of the most troublesome complications affecting diabetic patients and current therapy provides inadequate pain relief. In the present study, the effect of adenosine was investigated in a model of diabetic neuropathic pain. Diabetes was induced by streptozotocin (65 mg/kg, ip) in male Sprague Dawley rats and subjected to thermal (cold and hot) and chemical (formalin) stimuli. Diabetic rats developed hyperalgesia by the end of six weeks in thermal and chemical stimuli test. Adenosine (100, 200 and 500 mg/kg, ip) produced significant reversal of responses to thermal and chemical stimuli in diabetic rats. 8-Cyclopentyl-1, 3 dipropylxanthine (DPCPX 1 mg/kg, ip), an adenosine A1-receptor antagonist, but not 3,7-dimethyl-1-propargylxanthine (DMPX 1 mg/kg, ip), an adenosine A2A receptor antagonist, reversed the protective effect of adenosine. These results indicate that adenosine is an effective analgesics in a model of diabetic neuropathy, and the protection produced by adenosine is via stimulation of adenosine A1-receptors. PMID- 19552054 TI - 2-Deoxy-D-glucose alters the cardio-respiratory parameters in anaesthetized rats. AB - 2-Deoxy-D-Glucose (2-DG), a synthetic analogue of glucose, is used as an anticancer agent either alone or in combination with other tumor treatment protocols. The present study was conducted to identify the systemic effects of 2 DG on parameters of vital importance. The blood pressure, ECG and respiratory excursions were recorded in anesthetized adult rats. At the end (after 120 min) of experiments, the plasma glucose and serum insulin levels were estimated. Injection of 2-DG (0.5 g/kg) produced an immediate increase in mean arterial pressure (MAP) and respiratory rate. The increase in MAP continued throughout the period of observation (120 min) and the maximal increase was seen at 90 min (27%). Whereas, the respiratory rate decreased by 17% at 15 min which decreased further to 37% by 120 min. Heart rate also decreased after 2-DG in a time dependent manner and 40% decrease was observed at 120 min. Administration of 2-DG increased the plasma glucose level significantly (30%) as compared to saline control group but did not increase the serum insulin level. The results indicate that 2-DG alters the cardio-respiratory parameters by mechanisms unrelated to plasma insulin activity. PMID- 19552055 TI - Antinociception induced by central administration of histamine in the formalin test in rats. AB - In the present study, effects of intracerebroventricular (icv) administration of histamine, mepyramine (H1-receptor antagonist) and famotidine (H2-receptor antagonist) have been investigated on the formalin test in rats. Subcutaneous injection of formalin (50 microl, 1%) into the ventral surface of the left hind paw produced a marked biphasic pain response (first phase: 0-5 min and second phase: 15-45 min). All the performed treatments did not significantly influence the first phase of pain. Histamine at the doses of 10 and 40 microg and mepyramine and famotidine at the same doses of 20 and 80 microg, significantly (P < 0.05) decreased the late phase of formalin-induced pain. Pretreatments with mepyramine and famotidine at the same dose of 80 microg, significantly (P < 0.05) prevented the histamine (40 microg)-induced antinociception. These results indicate that brain histamine produces antinociception, and both central H1 and H2 receptors may involve in the histamine-induced antinociception in the formalin test in rats. PMID- 19552056 TI - Air pollution and fuel vapour induced changes in lung functions: are fuel handlers safe? AB - Automobile exhaust derived air pollutants have become a major health hazard. Coupled with the inhalation of fuel vapour, as occurs in petrol station workers, this may lead to significant impairment of lung function. Spirometric lung functions were studied in 58 petrol station workers to examine this possibility. The forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), forced expiratory flow 25%-75% (FEF25-75) and peak expiratory flow (PEF) were recorded and analysed separately for smokers and non-smokers. The workers were divided into 5 groups for analysis of data based on the number of years of work in the petrol pumps. Outdoor air analysis was also carried out. The FVC, FEV1 and PEF declined significantly with increasing years of work in petrol stations in both smokers and non-smokers. Smoking as an independent variable was found to affect the FEV1 significantly but not FVC or PEF. The FEF25-75 was found to be the most affected spirometric value with a significant reduction with increasing years of work. Smoking as such did not affect it. Oxides of nitrogen (NOx), suspended particulate matter (SPM) and particulate matter less than 10 microns (PM10) in outdoor air were higher than the national ambient air quality standards. Exposure to automobile exhaust and fuel vapour impairs lung function in a time-dependent manner. Cigarette smoking appears to accelerate the decline. PMID- 19552057 TI - Effect of 1 week '61-points relaxation training' on cold pressor test induced stress in premenstrual syndrome. AB - The present study was conducted on 50 clinically healthy women volunteers who were in their reproductive age group and in their premenstrual period. Thirty women having premenstrual syndrome (PMS) were compared with 20 control women to evaluate 1 week training of 61-points relaxation (61-PR) yogic exercise. In both the groups cold pressor test (CPT) was performed; and systolic blood pressure (SBP; mmHg), diastolic blood pressure (DBP; mmHg) and heart rate (HR/min) were measured. Basal SBP, DBP and HR of women with PMS was significantly higher than the control subjects with P value 1.2E-05, 0.000583 and 0.005707 respectively, suggesting the presence of stress in women with PMS. Immediately after CPT, the said parameters were statistically significantly more raised in PMS group than control group (P = 6.14E-05, 2.78E-06, 0.003799) respectively. Following 1 week of 61-PR training, the control group showed a significant decline in SBP, DBP & HR (P = 0.00769384, 0.00852894, 0.62316217) respectively. However, the PMS group showed a statistically highly significant reduction in SBP, DBP & HR (P = 2.55E 07, 4.35E-07 and 6.4904E-06) respectively immediately after CPT. These results suggest a reduction in sympathetic activity by 61-PR training and it can be used as an effective relaxation tool during premenstrual stress. PMID- 19552058 TI - Release of copper from CuT 380A co-incubated with human semen and its effect on sperm function in vitro. AB - Release of copper and its effect on functional integrity of human sperms in vitro were assessed following co-incubation of semen with CuT 380A. After 30 min of incubation with semen, release of copper ions from CuT 380A was found to be 9.2 to 40 times higher compared to control incubations with PBS. Sperm function tests, when simultaneously performed following loss of motility in sperms (> 95%) after 120 min of copper exposure, depicted a significant (P < 0.001) reduction in sperm viability and hypo-osmotic swelling (HOS) response. However, the affected sperm populations revealed no significant alterations in other functional tests like acrosomal status or nuclear chromatin decondensation. It is therefore concluded that the high release of copper from CuT 380A drastically lowers sperm motility, viability and HOS response but only marginally affects the acrosome status or nuclear chromatin condensation in short term incubations. PMID- 19552059 TI - Possible role of male factors in recurrent pregnancy loss. AB - OBJECTIVE: To evaluate various causes possibly contributing towards recurrent pregnancy loss (RPL), particularly male factors. Prospective study of 75 couples with history of RPL who were investigated for genetic, anatomic, immunological, infective and systemic causes in both partners. Functional sperm capacity was assessed by the Hypo-osmotic swelling test (HOS), Acrosomal Reaction (AR), Nuclear condensation-decondensation test (NCD) and Seminal Total Leukocyte Count (TLC) along with semen analysis. Twenty male volunteers with recently proven fertility were also included for detailed sperm morphology and sperm functions test as controls. Amongst male partners 3 (4%) had varicocele, 23 (30.6%) had infection, 1 (1.3%) immunological and 1 (1.3%) had genetic abnormality. Sperm motility, viability and sperm function tests were significantly lower in the RPL group as compared to the control group (P = 0.000). Male factor might be a possible contributing factor towards RPL. Both the partners should be evaluated and treated simultaneously in order to achieve desirable outcome. PMID- 19552060 TI - Oxidative stress in hypertension: association with antihypertensive treatment. AB - There is growing evidence that oxidative stress contributes to the pathogenesis of hypertension. Our aim was to measure oxidative stress in hypertensive subjects, and assess the potential confounding influences of antihypertensive therapy. Serum malondialdehyde and antioxidant levels were estimated in patients at the time of presentation and also after a antihypertensive therapy for 3 months. During the period of study no antioxidant/s was given to the patients and control subjects. Mean blood pressure values were altered in the hypertensive patients following antihypertensive therapy from their respective values observed at the time of presentation. Serum malondialdehyde levels were significantly higher in the hypertensive patients in comparison to control cases. The antioxidant activity of enzymes super oxide dismutase, glutathione and non enzymatic antioxidant levels of vitamins E and C were significantly lower in patients compared to controls. After 3 months of antihypertensive treatment all the above parameters showed reversal in the respective levels of serum malondialdehyde and antioxidant activity. Antihypertensive medications lower the blood pressure and thereby results in reduced oxidative stress which indicates that oxidative stress is not the cause, but rather a consequence, of hypertension. PMID- 19552061 TI - Association of anticardiolipin antibodies levels with instent restenosis in patients with coronary artery disease. AB - Present study was conducted to evaluate the association of IgG anticardiolipin antibodies with instent restenosis in patients having undergone percutaneous intervention with bare metal or drug eluting stents. Coronary artery disease patients with stent placement at least 6 months prior were screened for eligibility. 26 satisfied the inclusion/exclusion criteria. 10 patients with symptoms of restenosis, confirmed on check angiography served as cases and 16 without symptoms of restenosis served as control. Unpaired t- test was applied to ascertain the significance of any difference between control and study groups. Antibody levels were estimated on ELISA reader. The mean (+/- SD) anticardiolipin antibodies levels in cases and controls were 11.8 +/- 5.1 GPL/U/ml and 14.3 +/- 10.2 GPL/U/ml, respectively. The difference was not statistically significant (P > 0.05). In conclusion, we did not observe any significant correlation between the level of IgG aCL and instent restenosis. PMID- 19552062 TI - Effect of methylene blue on oxidative stress and antioxidant defence parameters of rat hepatic and renal tissues. AB - The objective of the present study was to investigate the effect of administration of 1 mM methylene blue (MB) in drinking water for 30 days on hepatic and renal antioxidant status in female adult Wistar strain rats (n = 5). MB failed to induce significant change in any of the measured antioxidant defence parameters namely, superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH). However, a marginally significant (P < 0.05) increase in the level of lipid peroxidation (LPx) was recorded in liver, while a reduction (P < 0.05) in its level in the kidney was noticed. Serum alanine amino transferase (AlaAT) and creatinine levels significantly (P < 0.001) decreased in MB treated rats without any change in blood urea nitrogen (BUN) level. Our findings suggest that the effect of MB as administered in the present study was tissue specific with regard to the level of LPx, however, in general, it does not impair liver and kidney functions as evidenced by serum parameters. PMID- 19552063 TI - Levels of malondialdehyde and antioxidants in the blood of patients with vitiligo of age group 11-20 years. AB - The aetiopathogenesis of vitiligo has not been fully understood. To investigate the role of oxidative stress in pathogenesis of vitiligo, we examined the blood level of antioxidants and malondialdehyde in 40 patients with generalized vitiligo and 40 healthy volunteers belonging to same age group. Our results revealed that the level of malondialdehyde was significantly raised while those of vitamin E, uric acid and ceruloplasmin were significantly lowered (P < 0.05) in patients with generalized vitiligo as compared to controls. This demonstrates the presence of an imbalance in the oxidant-antioxidant system in the blood of vitiligo patients and thus provides support for a free radical mediated damage as a pathogenic event in vitiligo. PMID- 19552064 TI - Effect of surgical stress on neutrophil function. AB - The present study was undertaken to find out the effect of surgical stress on nonspecific immune response. Twenty patients posted for various elective surgeries participated in the study (male : 17, female : 3, age : 43.4 +/- 2 yrs). The blood samples were taken preoperatively (4 to 6 days prior to surgery) and the following parameters were assessed: phagocytic index of neutrophils, avidity index of neutrophils and percentage of neutrophils in differential count. These were compared with the respective parameters assessed in the blood samples taken 24 hours after surgery. There was a significant (P = 0.0001) decrease in the phagocytic index of neutrophil and a significant (P = 0.003) increase in the percentage of neutrophils in differential count in the postoperative blood samples. However, the avidity index did not show a significant change. It could be tentatively concluded that surgical stress causes depression of nonspecific immunity in the early postoperative period. PMID- 19552065 TI - Metabolic syndrome--prevalence and significance of its detection in selected population in Pondicherry. AB - Obesity by itself is not an immediate lethal disease, however, it is a significant risk factor associated with a range of serious non-communicable diseases. Our aim is to find out the prevalence of metabolic syndrome in interns coming from higher socioeconomic class. Sixty one interns participated in the study. BMI, WHR and blood chemistry were performed. EGIR criteria were considered to identify the metabolic syndrome. According to EGIR criteria, insulin resistance was found in 24.5% cases and metabolic syndrome exists in 14.7% of overweight/obese interns. Insulin resistance as per HOMA-IR was found in 6% of those having metabolic syndrome. High diastolic blood pressure, low HDLc are prone to develop metabolic syndrome by six times where as an increase in WHR (central obesity) particularly in male are eleven times more prone to develop metabolic syndrome. PMID- 19552066 TI - Pathophysiological approach to the management of scorpion envenomation. AB - Indian red scorpion (Mesobuthus tamulus; MBT) produces lethal stings and is a matter of concern in certain parts of India. MBT envenomation produces multi systemic involvement, thus presents difficulty in the management. Symptomatic treatment has been practiced earlier that failed to relieve the toxic effects of the venom. Therefore, present manuscript deals with pathophysiologically based approach in the management of toxicity considering the merits and the demerits of treatment protocols so as to evolve a consensus in the treatment strategies of scorpion envenomation. PMID- 19552067 TI - Ethanol intoxication--dose dependent effect on circulating testosterone level in Wistar rats. PMID- 19552068 TI - Temperature effect on behaviour, oxygen consumption, ammonia excretion and tolerance limit of the fish fingerlings of Alepes djidaba. AB - The present study has been carried out to determine the effect of temperature on behaviour, oxygen consumption, ammonia excretion and tolerance limit of the fish fingerlings of Alepes djidaba, which were collected at Kalpakkam. The fish fingerlings were placed at different temperatures, based on the thermal tolerance limit of fish and thermal outfall of the Madras Atomic Power Station (MAPS). The thermal tolerance experiments were conducted in two ways: in direct exposure and in gradually increasing temperature for duration of 48 hr. The upper and lower lethal temperatures for the fish fingerlings of Alepes djidaba were 38.5 degrees C and 14 degrees C respectively. During tolerance experiment, no mortality was observed at 33 degrees C and 35 degrees C. But at 38 degrees C with gradual increase in temperature, 100% loss of equilibrium was observed at 37.7 degrees C in 36 min and mortality was observed at 38 degrees C in 55 min. On the contrary, when the fish fingerlings A. djidaba were directly exposed to 38 degrees C, almost 100% loss of equilibrium and mortality were recorded in 15 min and 31 min respectively. At 40 degrees C with gradual increase in temperature, 100% loss of equilibrium was recorded at 38.5 degrees C in 46 min and mortality was recorded at 39 degrees C in 50 min. On the other hand, when the fish fingerlings of Alepes djidaba were directly exposed to 40 degrees C, 100% mortality occurred immediately within one minute. These behavioral responses include an elevated temperature of deltaT 10 degrees C, surfacing, dashing against glass wall, jumping out of the water, etc. In general, the rate of oxygen consumption and ammonia excretion was found to enhance with increasing temperature. The oxygen consumption was found to increase from 0.97 mg O2/g/hr at 30 degrees C to 2.2 mg O2/g/hr at 35 degrees C. Similarly, the excretion of ammonia also increased from 3.18 microg/g/hr at 30 degrees C to 5.91 microg/g/hr at 35 degrees C. In the present study, it was found that gradual increase in temperature favours the fish population to escape from the thermal exposure as compared to direct exposure, and also the temperature impact due to thermal discharge from MAPS is only minimal (35 degrees C) and no mortality could be caused to the fish population. PMID- 19552069 TI - Impact of chlorination on the incidence of cancers and miscarriages in two different campus communities in India. AB - Long-term impacts of drinking chlorinated water on the incidence of cancers and miscarriages were assessed in a population-based cross-sectional study conducted in the two campus communities of IIT Kanpur (IITK) and IIT Kharagpur (IITKgp). IITK has been using untreated groundwater since the community was established in 1963, while IITKgp has been using chlorinated water for more than 30 years. A house-to-house survey was carried out to gather information on residential history, i.e age, education, income, source and extent of treatment of water and health characteristics. Only adults above 20 years of age were included for data analyses. Odds ratios were calculated based on the hypothesis that exposure to chlorinated drinking water may result in a higher incidence of cancers and miscarriages as found in many studies. The odds ratios (OR) in this study were found to be 0.56 (95% CI = 0.16 to 1.62) for cancers and 0.33 (95% CI = 0.19 0.56) for miscarriages. These OR values are not statistically significant indicating the lack of association between cancers or miscarriages and exposure to chlorinated drinking water, and are in agreement with some published epidemiological studies as well. Reciprocal OR values were calculated based on an alternative hypothesis that chlorination actually decreases the risk of cancers and miscarriages. Based on this, the OR values for cancers are 1.77 (95% CI = 0.55 to 5.66) and for miscarriages are 3.07 (95% CI = 1.78 to 5.29). These results show that there is no association between exposure to chlorinated drinking water and cancers while there is significant decrease in the incidence of miscarriages for those exposed to chlorinated drinking water. PMID- 19552070 TI - Quality of ground water used for irrigation in Ujjain district of Madhya Pradesh, India. AB - The ground water samples collected from different tehsils of Ujjain district were analysed in the laboratory for electrical conductivity (EC), pH, cations (Na+, K+, Ca2+ and Mg2+) and anions (CO3(2-)), HCO3-, Cl- and SO4(2-)). The water samples were categorized as per criterion developed by Central Soil Salinity Research Institute, Karnal, Haryana, India. Out of 712 samples, 105, 144, 150, 84, 68, 111 and 50 samples belong to Ujjain, Mahidpur, Khachrod, Tarana, Barnagar, Nagda and Ghatia tehsils of the district respectively. 80% samples indicated good water quality at the district as a whole, whereas 14% were found saline (marginally saline (B1)-9%, saline (B2)-4%, high SAR saline (B3)-1%) and 6% as alkali (marginally alkali (C1)-5%, and alkali (C2)-1%) categories. In general, these waters were of Ca-Na-Mg type with the dominance of Cl followed by HCO3 and CO3 in case of good quality waters. The poor water quality belonged to categories of B1, B2, B3, C1 and C2, which were either having accumulation of salts (high EC) or high sodium adsorption ratio (SAR) and residual sodium carbonate (RSC). High significant negative correlation coefficient between pH and SO4-- was observed. The sodium (Na+) content showed significant positive correlation coefficient with EC. Similarly, HCO3- showed a significant positive correlation with RSC content. The ground water quality map of the district was generated using ERDAS-IMAGINE 8.7 Software showing different categories. PMID- 19552071 TI - Ground water quality in Coimbatore, Tamil Nadu along Noyyal River. AB - The ground water quality in Coimbatore city along the Noyyal river during pre monsoon and post-monsoon seasons in 2005 is discussed in this paper. Water samples were collected from 12 wells on either side of the river. The physico chemical analysis of the collected samples was carried out for the parameters, such as pH, total dissolved solids (TDS), electrical conductivity (EC), total hardness (TH), alkalinity, calcium (Ca2+), magnesium (Mg2+), chloride, sulphates (SO4(2-)) and sodium (Na+) . From the study, it is revealed that EC, Ca2+, Mg2+ have high concentrations in pre- and post-monsoon seasons. In most of the places, the quality of water was not found suitable for irrigational and industrial activities because of high concentrations of electrical conductivity and total hardness. In general, the values of all the studied parameters were high and above the standards during the post-monsoon season, compared to the pre-monsoon season in 2005. PMID- 19552072 TI - Effect of inoculum-substrate ratio on acclimatization of pharmaceutical effluent in an anaerobic batch reactor. AB - Anaerobic treatment has gained tremendous success over the past two decades for treatment of industrial effluents. Over the past 30 years, the popularity of anaerobic wastewater treatment has increased as public utilities and industries have utilized its considerable benefits. Low biomass production, row nutrient requirements and the energy production in terms of methane yield are the significant advantages over aerobic treatment process. Due to the disadvantages reported in the earlier investigations, during the past decade, anaerobic biotechnology now seems to become a stable process technology in respect of generating a high quality effluent. The objective of the present experimental study was to compare the biodegradability of recalcitrant effluent (pharmaceutical effluent) for various inoculum-substrate ratios. The batch experiments were conducted over 6 months to get effect of ratio of inoculum substrate on the acclimatization of pharmaceutical effluent. The tests were carried out in batch reactors, serum bottles, of volume 2000 mL and plastic canes of 10000 mL. Each inoculum was filled with a cow dung, sewage and phosphate buffer. The batch was made-up of diluted cow dung at various proportions of water and cow dung, i.e., 1:1 and 1:2 (one part of cow dung and one part of water by weight for 1:1). The bottles were incubated at ambient temperature (32 degrees C 35 degrees C). The bottles were closed tightly so that the anaerobic condition is maintained. The samples were collected and biodegradability was measured once in four days. The bottles were carefully stirred before gas measurement. The substrate was added to a mixture of inoculum and phosphate nutrients. The variations in pH, conductivity, alkalinity, COD, TS, TVS, VSS, and VFA were measured for batch process. The biogas productivity was calculated for various batches of inoculum-substrate addition and conclusions were drawn for expressing the biodegradability of pharmaceutical effluent on acclimatization period and influent COD concentration. PMID- 19552073 TI - Simultaneous determination of trace heavy metals in ambient aerosols by inductively coupled plasma atomic emission spectrometry after pre-concentration with sodium diethyldithiocarbamate. AB - The simultaneous determination of heavy metals associated with airborne particulate matter in the atmosphere of the city Isfahan (Iran) was performed by inductively coupled plasma atomic emission spectrometry (ICP-AES) after pre concentration with sodium diethyldithiocarbamate. The preconcentration procedure developed found instrumental to determine the trace heavy metals associated with ambient aerosols collected at a short sampling period or collected from rural areas where the concentrations of these metals are much less than those in urban areas. Several samples were analyzed by both flame atomic absorption spectrometry (FAAS) as a conventional method and the proposed method. The results obtained by the two methods were found in good agreement. The method was applied to the determination of atmospheric level of heavy metals in rural area and also for study of variation in levels of heavy metals in urban atmosphere during the days and nights. PMID- 19552074 TI - Adsorption of oils, heavy metals and dyes by recovered carbon powder from spent pot liner of aluminum smelter plant. AB - Aluminum smelter plants employ Hall-Heroult electrolysis cells for electrolysis of molten cryolite to recover aluminum metal by electrolysis. These cells use carbon cathode blocks as a lining material inside. At the end of service life of the cells, pot lines are discarded and new carbon blocks are laid for fresh charging. These used carbon cathode blocks, known as spent pot liners, are heavily infested with toxic elements such as fluoride, cyanide, alkali, etc. Therefore, their disposal in open field poses great environmental risk. A simple process has been developed for decontamination of these spent pot liners and to recover its carbon value. The experiments indicated that this carbon, in the form of fine powder (around 20 micron in size) can absorb toxic elements like heavy metals, dyes, oils, etc. to a great extent and thus can be used for mitigating environmental pollution occuring due to various toxic wastes. PMID- 19552075 TI - Environment-friendly cycle time optimization and quality improvisation using Six Sigma. AB - Healthy environment in any organization can make a difference in improving productivity and quality with low defect, lack of concentration, willingness to work, minimum accidental problems etc. Six Sigma is one of the more recent quality improvement initiatives to gain popularity and acceptance in many industries across the globe. It is an alternative to TQM to obtain minimum manufacturing defect, cycle time reduction, cost reduction, inventory reduction etc. Its use is increasingly widespread in many industries, in both manufacturing and service industries with many proponents of the approach claiming that it has developed beyond a quality control approach into a broader process improvement concept. PMID- 19552077 TI - Polynomialized model for critical DO deficit. AB - The existing DO sag models of Streater-Phelps have become obsolete in the present day context of polluted streams in which a part of the BOD removal necessarily takes place through sedimentation. The Streater Phelps models do not consider this aspect. Bhargava's theoretically derived model for the critical DO deficit rests on an almost linear removal of the settleable BOD and an exponential decay of non-settleable BOD. However, the Bhargava's model has two independent but complex expressions, one each for times less than and greater than the transition time. A polynomialized form of Bhargava's models for critical DO deficit has been developed as a single expression and universally applicable without any regard to the transition time. Unlike, the Streater-Phelps or Bhargava models, the presented polynomial form of Bhargava models, for critical DO deficit has an additional advantage of evaluating the critical dissolved oxygen deficit concentrations directly and without first determining the time of occurrence of such a deficit. The material presented would thus add to the exiting literature on the subject. PMID- 19552076 TI - Multivariate analysis of groundwater resources in Ganga-Yamuna basin (India). AB - Groundwater quality data on physico-chemical, bacteriological and heavy metal concentrations in three cities (Faridabad, Allahabad and Varanasi) in Ganga Yamuna basin was subjected to multivariate analysis (MVA) using SPSS. The factors extracted showed high loading (> 0.3) of various parameters, such as Cl, conductivity, TDS, hardness, Na, Mg, and SO4, indicating contamination due to leaching of pollutants. Major manifest variable associated with these factors is the unorganized solid waste dumping practiced in all the cities. Bacterial contamination of hand pump samples in Allahabad is attributed to surface water groundwater interaction. The factor with high loading of Ca and F is indicative of geological conditions of the region. Wells in Yamuna river sub-watershed exhibit less freshwater recharge, which is attributed to surface water pollution and sediment deposition in the river. Thus, the methodology for hydrogeological analysis is useful to identify critical water quality issues and possible sources of pollution in river basins. PMID- 19552078 TI - Applicability of liquid ion exchange to alum recovery from waste stabilization pond sludge. AB - This paper reports the evaluation of the tertiary application of relatively low doses of alum for improving the overall performance and reliability of existing wastewater treatment plants. With this type of system, the additional capability for phosphorus removal is imbibed by simply altering the chemical dose. The study was conducted on wastewater from a Stabilization pond having capacity of 54 mld, which was treating municipal wastewater from Ujjain city. The wastewater had suspended colloidal impurities. PMID- 19552079 TI - Energy recovery from municipal solid waste in an anaerobic reactor. AB - Anaerobic digestion of municipal solid waste was carried out in the laboratory at room temperature to assess the bio-energy production from municipal solid waste (MSW) with high total solids content. The total biogas production from the municipal garbage was found to be 3.2 L in 120 days. The results from the biomethanation process showed that an increase in gas production was observed with increase in digestion period when the bioconversion parameters were found to be favorable for the production of gas. Changes in the parameters, such as pH, affected the production of gas significantly. Samples taken from the reactor at definite interval of time during the degradation process showed considerable reduction in total volatile solids, total carbon, total nitrogen and COD, etc. indicating the waste stabilization. PMID- 19552080 TI - Role of the fungus--Periconiella sp. in destruction of biomedical waste. AB - The disposal of biomedical waste is a burning problem in developing countries due to scarcity of resources and funds in view of its high cost involved. In this paper, a very cheap, easily available and effective method by using fungus- Periconiella sp. isolated from cow dung was successfully tried, which degrades the biomedical waste materials very efficiently and quickly without leaving any trace of harmful effect on the population. PMID- 19552082 TI - Medical informatics --power of print and vision. PMID- 19552083 TI - Variations of adolescent blood pressure by multifactorial analysis in an urban slum of Kolkata. AB - To find out the factors responsible for variations of adolescent blood pressure, a community-based cross-sectional study was undertaken among 1081 adolescents, aged 10-19 years, selected by simple random sampling in an urban slum of Kolkata, Chetla in 2003-04. Age, educational status, additional ghee/butter intake with diet, family history of hypertension, weight, height and body mass index were found to be associated with blood pressure in unifactorial analysis, hence these variables were considered for multifactorial analysis. All these independent variables were correlated with both systolic and diastolic blood pressure except additional ghee/butter intake with food. In stepwise multiple regression analysis, age, education, family history of hypertension, weight, height and body mass index contributed 64.6% of the total variation of systolic blood pressure, where weight alone contributed to 62.6% while 35.4% of variations remained unexplained. In case of diastolic blood pressure, 56.8% of the total variations were due to age, education, family history of hypertension, weight, height and body mass index, out of which weight alone contributed 56% while remaining 43.2% of total variations remained unexplained. Since single most important predictor of blood pressure was weight, regular aerobic physical activity, dietary modification, behavioral changes and health education are recommended for weight reduction and prevention of obesity. PMID- 19552081 TI - Nutrient removal by root zone treatment systems: a review. AB - The Root Zone Treatment System (RZTS) has been used widely for nutrient removal in European countries. In spite of having its more adaptability in tropical region like India its use to address nutrient induced issues in the country is very less. The lack of widely accepted data, non consensus of scientists over nutrient removal mechanism and inability to apply performance standards observed in other countries directly might have hampered the acceptance of this technology in India. A few technology assessment programs are being conducted in collaboration with other countries to engineer this technology but nutrient removal aspects are not essentially focused. In this context, there is need to direct lab scale research to identify potential wetland plants, bed media and comparative study of their combination specific performance under similar conditions. The field application of the data will help to understand variability in performance and disparities in the mechanism. The systems would be amended based on these studies to establish combination specific performance standards for typical Indian conditions. Maintenance strategy and optimization of design will help to foster the technology. The development strategy should give due consideration to the contributions of other countries so as to avoid repetition of work which will save time, money and efforts, and help for the real acceptance of RZTS in Indian conditions. PMID- 19552084 TI - Acute pancreatitis in Goa--a hospital-based study. AB - A prospective analysis of the epidemiology and outcome of patients admitted with acute pancreatitis to a tertiary health care centre in Goa was carried out during the time period of 1st June, 2003 to 31st January, 2005. The patients studied were those who were admitted to the Goa Medical College with a diagnosis of acute pancreatitis based on a serum amylase of greater than 180 Somogyii units with appropriate clinical and radiographic evidence. The selection criteria were fulfilled by 282 patients. Acute pancreatitis accounted for 2.29% of all admissions and 4.9% of all deaths in the department of surgery. The disease was seen to affect males more commonly (96.1%), alcohol, being the predominant (92.2%) aetiological factor. The median age for occurrence of the disease was 40 years. Severe acute pancreatitis was encountered in 32.9% cases with a mortality rate of 12.05%. Mortality was higher in patients older than 50 years. Complications included pseudocysts (n = 34), abscess (n = 2), necrosis (n = 6), ascites (n = 13), acute respiratory distress syndrome (n = 13), acute renal failure (n = 14), shock (n = 14), coagulopathy (n = 1) and pleural effusion (n = 26). The widespread availability and use of locally made cheaper varieties of alcohol in our geographical location explains the trend towards alcoholic pancreatitis and younger age groups being affected by the disease. PMID- 19552085 TI - Prevalence of microvascular complications in metabolic syndrome in comparison to type 2 diabetes mellitus. AB - To identify the prevalence of microvascular complications in patients with metabolic syndrome and compare them with patients with only diabetes mellitus, a retrospective study was carried out at a tertiary health care centre in South India with 150 patients, in two groups of 75 with almost identical age distributions. The test group (n = 75) fulfilled the NCEP AT III guidelines for metabolic syndrome. The control group (n = 75) had only diabetes mellitus and no other components of metabolic syndrome. They were assessed for the microvascular complications of diabetes mellitus, namely retinopathy (by direct ophthalmoscopy and zeiss f450 plus fundus camera), nephropathy (using overnight spot albumin/creatinine ratio) and neuropathy (using clinical features, abnormal results on Achilles tendon reflex and sensory perception by 10 g Semmes Weinstein monofilament). In the test group, 33.3% were found to have retinopathy, while in control group 13.3% had retinopathy; 38.7% in the test group and 22.7% in the control group had nephropathy; 50.7% in the test group and 26.7% in control group had neuropathy. On comparison the test group had statistically significant (retinopathy p < 0.05, nephropathy p < 0.001 and neuropathy p < 0.05) increase in the prevalence of microvascular complications as compared to the control group. Patients with metabolic syndrome have more prevalence of microvascular complications as compared to patients with only diabetes mellitus. PMID- 19552086 TI - A study of comparison of efficacy and safety of talc and povidone iodine for pleurodesis of malignant pleural effusions. AB - Pleurodesis is considered as the best palliative therapy for the treatment of symptomatic malignant pleural effusion. Several chemical agents are used for this purpose with variable efficacy and safety. The present study is to compare the effectiveness and safety of talc and povidone iodine as chemical agents for pleurodesis in patients of malignant pleural effusion. Fifty-two patients with malignant pleural effusion admitted in the department of chest of Calcutta National Medical College, Kolkata were selected for the study. Pleurodesis with povidone iodine and talc in slurry was done in 28 and 24 patients respectively. Efficacy and safety of these agents were assessed during a follow-up period of six months. Among the 52 patients, 42 were males and 10 females. Age ranged from 40 to 64 years with mean age of 56.4 years. In 41 patients effusion was secondary to bronchogemic carcinoma, 8 had effusion secondary to breast carcinoma, 1 had effusion due to non-Hodgkin's lymphoma, while primary malignancy was unknown in 2 patients. Among the 24 patients treated with talc pleurodesis, 20 had bronchogenic carcinoma, 3 had breast carcinoma and 1 had unknown primary malignancy. Out of the 28 patients treated with povidone iodine pleurodesis, bronchogenic carcinoma was present in 21 patients, breast carcinoma in 5 patients, non-Hodgkin's lymphoma and unknown primary malignancy was present in 1 patient each. Pleurodesis with talc showed complete success in 19 patients, partial success in 3 patients and failure in 2 patients. Pleurodesis with povidone iodine showed complete response in 24 patients, partial response in 1 patient and failure in 3 patients. Chest pain occurred in 4 patients of talc pleurodesis and 5 patients of povidone iodine pleurodesis, 3 patients of each group had fever. There was no death in the peripleurodesis period. During the 6 months follow-up, 12 patients of talc pleurodesis and 18 patients of povidone iodine pleurodesis died. Talc is slurry and povidone iodine is equally effective and safe pleurodesing agent for symptomatic malignant pleural effusion. However povidone iodine can be preferred option because of easy availability and low cost. PMID- 19552087 TI - Rheumatological manifestations in diabetes mellitus. AB - A study was undertaken in the department of medicine from November 2003 to July 2005 with a view to find out the prevalence of different rheumatological problem in Indian diabetic population. Although several such studies have been conducted in western population very limited data are available from our country. A total of 80 patients were studied out of which 43 had some form of rheumatological manifestations. Dupuytren's contracture was found in highest number of cases (n = 23) followed by limited joint mobility in 16 patients; adhesive capsulitis in 19 patients; trigger finger in 4 patients; aglodystrophy, carpal tunnel syndrome and hyperostosis were found in 2, 3 and 2 cases respectively. Symptomatic osteo arthritis was found in 19 cases. PMID- 19552088 TI - IMA GFATM RNTCP PPM: gearing up to face ground realities. AB - To fight the menace of tuberculosis public-private mix model as being nurtured by Central TB Division and IMA may become an appropriate model for the developing and even for developed countries. IMA had sensitised 12147 private practitioners in five target states in India. Also 592 private practitioners have been trained in District Training Programmes. Frequently asked questions in these sensitisation and District Training Programmes are narrated in this article. The whole of India has been covered under DOTS strategy. Case detection rate of 70% is the key factor for the programme to succeed. CTD is able to reach case detection of new sputum positive (NSP) cases up to 55-60%. IMA is expected to fill the gap to reach magical figures of 70% NSP case detection rate. The risk of active tuberculosis with latent infection is increased 100-fold by HIV infection. Treating HIV and tuberculosis together and initiative of HIV treatment as been elaborated. Immune reconstitution inflammatory syndrome (IRIS) is characterised by a clinical worsening of a known condition or the appearance of a new condition after initiating antiretroviral therapy (ART). Details about IRIS have been narrated in this article. PMID- 19552089 TI - A case of rhino-orbital cerebral mucormycosis with diabetic keto-acidosis. AB - A 60-year-old lady was admitted in the hospital with the complaints of burning sensation during micturition and abdominal pain. She was diagnosed to have moderate hydronephrosis with left lower 1/3rd ureteric calculus for which ureteroscopy and lithotripsy were done. Her pre-operative random blood sugar was normal. On the 7th postoperative day the patient developed diabetic keto-acidosis which was followed by an acute onset of right sided peri-orbital oedema, proptosis and facial pain. Subsequently she developed diminished vision and lower motor neurone type of Ill, IV and VI cranial nerves paralysis on right side, disorientation and minimal left sided hemiparesis. ENT examination revealed black eschar nasal turbinates, nasal septum and palate and a provisional diagnosis of rhino-orbital cerebral mucormycosis was made. Extensive debridement was done for the patient and the specimen culture showed growth of mucor species. Patient was started on intravenous amphotericin-B and she started improving dramatically. This case of rhino-orbital cerebral mucormycosis with diabetic keto-acidois is presented here for its rarity. PMID- 19552090 TI - Pelvic tuberculosis in a postmenopausal woman mimicking ovarian malignancy--a clinical dilemma. AB - A 72-year-old postmenopausal woman presented with dyspepsia, weight loss, abdominal pain, and ascites. Ultrasonography revealed a pelvic mass and evidence of pyometra. Serum CA-125 was raised. Paracentesis revealed lymphocytic exudate but no malignant cells or acid-fast bacilli. However, after drainage of pyometra endometrial curettings revealed epithelioid granuloma with acid-fast bacilli. She had complete recovery with full course of antituberculosis treatment. Thus, pelvic tuberculosis should be considered in the differential diagnosis of ovarian malignancy, which can prevent many unnecessary laparotomies. PMID- 19552091 TI - Primary mediastinal yolk sac tumour in a young female. PMID- 19552092 TI - Evaluation of sensitivity of different organisms to cefepime and tazobactum (megapime XP) in comparison to cefepime and ceftazidime. AB - The present study was carried out to evaluate the sensitivity of different organisms to cefepime and tazobactum (megapime XP) in comparison to cefepime and ceftazidime. Samples were collected from clinical specimens and micro-organisms were isolated from clinical specimens. Strains of micro-organisms isolated from clinical specimens were identified using standard methods like morphology, colony characteristics and biochemical reactions. Sensitivity was carried out using Kirby Bauer disc diffusion method. Minimum inhibitory concentrations (MICs) for all the antibiotics were carried out as per guideline of Clinical and Laboratory Standard Institute (CLSI). The results of the study demonstrated that cefepime and tazobactum exhibits good activity against Gram +ve and Gram-ve organisms. PMID- 19552093 TI - Evaluation of safety and efficacy of telmisartan-amlodipine combination in treating hypertension. AB - The objective of this open, non-comparative, prospective postmarketing surveillance (PMS) study was to identify, validate and quantify the safety and efficacy associated with the use of fixed dose combination (FDC) of telmisartan 40 mg + amlodipine 5 mg (T40+A5) in hypertensive patients with or without concomitant diabetes. The data was collected from 72 centres from all over India during the period of June 2007 to February 2008. A total of 251 patients of either sex and those who were newly diagnosed stage II hypertension, or those who were uncontrolled on monotherapy with or without diabetes mellitus were included in this study. Patients were prescribed with T40+A5 combination orally. Systolic BP (SBP), diastolic BP (DBP) and heart rate (HR) were measured at the start and at the end of 2, 4 and 8 weeks of treatment. Primary efficacy end points were reduction in clinical SBP/ DBP from baseline to study end and number of patients achieving JNC VII goals. Tolerability was assessed by treatment-emergent adverse events. Out of 251 patients, 208 patients had completed the study (120 males and 88 females), 42 were lost to follow-up the study and one patient was withdrawn due to adverse effects. The mean age of the patients was 54.5 +/- 0.98 years for males and 52.94 +/- 1.078 years for females. Diabetes mellitus was seen in 64.9% of cases, dyslipidaemia in 2.88%, previous IHD in 7.2% cases and chronic obstructive pulmonary disease (COPD) in 0.50% of cases. Reduction in the mean SBP was found to be 12.08%, 18.92% and 22.90% at the end of 2, 4 and 8 weeks respectively (p < 0.001). Reduction in the mean DBP was found to be 10.09%, 14.55% and 17.19% at the end of 2, 4 and 8 weeks respectively (p < 0.001). At the end of the study it was found that 86.3% of the hypertensive patients and 70% diabetic hypertensive patients achieved the JNC VII recommended goals. The overall incidence of ADRs was 7.69% with headache (1.92%) and vertigo (1.44%), as the commonest side-effect. According to physician's assessment of efficacy and tolerability 99.5% of total cases showed good to excellent response. In the treatment of stage II hypertensive patient the FDC of T40+A5 (Telar-AM) was found to be significantly effective in the reduction of SBP as well as DBP. Overall incidence of ADRs was lower and FDC of T40+A5 is well tolerated. PMID- 19552095 TI - National Vector Borne Disease Control Programme--a glimpse. PMID- 19552094 TI - Chiral non-steroidal anti-inflammatory drugs--a review. AB - A chiral centre is noted in majority of the NSAIDs. For NSAIDs the enantiomer with S configuration almost exclusively possesses the ability to inhibit prostaglandin activity. R-enantiomers of NSAIDs have poor COX inhibitory activity. Some R-enantiomers are not inert, and many have different actions. The R- to S- chiral inversion varies with biological factors and property of NSAID. The S- to R- chiral inversion is rare for all the NSAIDs. Various preclinical and clinical studies have demonstrated that chirally pure NSAIDs like dexketoprofen, dexibuprofen and S-etodolac are more potent than their respective R enantiomers. Favourable pharmacokinetic and pharmacodynamic profile of dexketoprofen, dexibuprofen and S-etodolac make them effective and well tolerated drug for the treatment of painful inflammatory conditions at half doses of recemate. Thus chiral switch of NSAIDs is a rational approach for the treatment of painful inflammatory conditions. PMID- 19552096 TI - Malaria associated acute renal failure--experience from Rourkela, eastern India. AB - Acute renal failure is a significant complication of falciparum malaria and is associated with increased morbidity and mortality. It occurs rarely in children and is seen frequently among the adults. Malaria associated renal failure may occur as a sole complication or as a component of multiple complications. Some of the patients may have normal urination (non-oliguric) and usually have better prognosis even without renal replacement therapy. Only a few research studies are available from Indian subcontinent on malarial acute renal failure. The present study is a hospital based study from eastern India. Rourkela is situated in the western part of Orissa which contributes a large number of falciparum malaria cases. The study was conducted at the internal medicine department of Ispat General Hospital. The clinical presentation of malaria patients in 2001 were analysed with special emphasis on malarial acute renal failure. The difference between patients with or without renal failure was compared. Seven hundred eighteen patients admitted to Ispat General Hospital in 2001 above the age of 14 years were analysed. Of these, 84 (11.8%) had serum creatinine >3 mg/dl. Seventy five patients were referred from different hospitals outside the township. The presenting complaints were fever (95%), oliguria (55%); loose motions (25%), and vomiting (51%). Headache was present in only 20% patients. Similarly, hypotension was encountered in about a third. Associated complications were significantly more common among patients having renal failure viz, Jaundice (77 versus 19%; p < 0.001), Cerebral malaria (59 versus 11%; p < 0.001), and hypoglycaemia (p < 0.05). The mortality in presence of acute renal failure was high (p < 0.001). Though malaria renal failure is a burning issue, still scant data is available in the literature, including India. The present study is an attempt to study the patients admitted to a referral hospital. The reason for high mortality is due to presence of multiple complications. The present study indicates that the presence of acute renal failure and jaundice together adversely influences the mortality. Hence, studies may be carried out to find out the reason of this changing trend as well as the methods to ameliorate/manage the situation. PMID- 19552097 TI - Does metformin augment the ovulation inducing effects of clomiphene in non-obese women with polycystic ovary syndrome? AB - To decide if metformin augments clomiphene response for ovulation induction in non-obese women with polycystic ovary syndrome, a prospective randomised placebo controlled trial was undertaken among 27 patients, who were found eligible for this study following clinical assessment and basic investigations. Women with polycystic ovary syndrome, even when non-obese, has increased insulin-resistance and do not respond favourably when treated with clomiphene for ovulation induction. The hypothesis is that by improving insulin- resistance and thus reversing from hyperinsulinaemia towards normal insulin secretion will enhance the responsiveness of these women to ovulation inducing effects of clomiphene. In the metformin group there were 15 patients who were given metformin 500mg orally 8 hourly daily for initial 3 months whereas in the placebo group there were 12 patients who were given folic acid (as placebo) 5mg orally once daily for initial 3 months. Thereafter, all the 27 patients in both groups were treated with clomiphene 50mg orally once daily from day 2 for 5 days in each month for subsequent 3 months. However, metformin and folic acid was continued in the metformin group and placebo group respectively for these subsequent 3 months when these patients were being treated with clomiphene for ovulation induction. Ovulation, as the outcome measure, was assessed by serial transvaginal ultrasound scanning from day 8/ day 9 and serum progesterone estimation on the 7th or 8th day following the ultrasound evidence of ovulation. Ovulation is taken to have occurred when serum progesterone was > or =8 ng/ml. Ovulation was noted to have occurred in 71.11% of the 45 cycles studied in 15 patients in the metformin group whereas ovulation occurred in 11.11% of the 36 cycles studied in 12 patients in the placebo group. This difference was highly statistically significant (p < 0.001) by Fisher's exact test. It can be conducted that metformin augments the ovulation inducing effects of clomiphene in non-obese women with polycystic ovary syndrome. PMID- 19552098 TI - Short-term mortality and complications in ST elevation myocardial infarction--the Heart Hospital experience. AB - The mortality rate of acute myocardial infarction has declined considerably in the past three decades. In view of paucity of literature from different centres from India on this issue, the present study was undertaken to determine the in hospital mortality with acute ST segment elevation myocardial infarction presenting to a tertiary care cardiac centre in India. Consecutive patients (n=862) with the diagnosis of acute ST segment elevation myocardial infarction admitted in Heart Hospital, Patna between June 2003 and July 2006 were included in this study. The in-hospital mortality and event rates (reinfarction, recurrent angina and heart failure) were analysed. The mean age of study population was 56 +/- 13 years. There were 690 males (80.05%) and 172 females (19.95%); 468 patients (54.29%) had hypertension, 384 patients (44.55%) were diabetic, 415 (48.14%) were smokers/tobacco chewers and 154 patients (17.86%) had past history of myocardial infarction. Anterior wall infarction was present in 435 patients (50.46%), 408 patients (47.33%) had inferior wall infarction, 115 patients (13.34%) had associated right ventricular or posterior wall infarction and 19 (2.20%) had antero-inferior infarction; 346 patients (40.14%) received thrombolytic therapy while the other patients were not thrombolysed due to various reasons (usually late arrival). The mean duration between symptom onset and hospital admission was 29.2. +/- 10.8 hours in the entire group (8.6 +/- 2.8 hours in the thrombolysed group). Of the total 862 patients, 107 patients (12.41%) died during in-hospital stay while 755 patients were discharged from the hospital in stable condition after a mean stay of 7.1 +/- 1.8 days. The in hospital mortality rate of acute ST segment elevation myocardial infarction in this study was 12.41%, which is comparable to reports from the west. However the revascularisation rate (thrombolysis or PTCA) remained low and most patients received thrombolysis late. PMID- 19552099 TI - A study on endocrine dysfunction in thalassaemia. AB - The aim was to study the prevalence and severity of hormonal imbalance affecting growth, gonadal and thyroid function in thalassaemic patients and to find out whether any correlation exists between the degree of tissue iron-overload and several patients characteristics like age, gender, foetal haemoglobin (HbF) level, type of thalassaemia (beta or E-beta), and the presence of specific endocrine abnormality. Sixty-eight consecutive non-chelated, transfusion dependent patients of beta and E-beta-thalassaemia with significant tissue iron overload (serum ferritin more than 2000 microg/l) were included. Standing height was noted and clinical features of hypogonadism were recorded. Insulin tolerance test was done to assess growth hormone reserve. Serum oestradiol, T3,T4,TSH were measured in fasting clotted sample, while pooled sera (from 3 consevutive morning samples) was used for testosterone, FSH and LH. Hypogonadism was the commonest abnormality, both in males (52.28%) and females (35.89%) followed by growth retardation (20.58%) and reduced growth hormone reserve (7.35%). There was no significant difference in the prevalence of endocrine dysfunction with regard to patient's age, gender, type of thalassaemia (beta or E-beta) amd HbF level. Hypogonadic females had a significantly elevated mean serum ferritin level. Subclinical hypothyroidism was present in 23.52% of patients, related to the duration of disease. No association was found between pituitarty and thyroid dysfunction. PMID- 19552100 TI - Epidemiology of Japanese encephalitis in context with Indian scenario. AB - Japanese encephalitis (JE) has been prevalent in various countries of East and South-East Asia since long. In India, JE virus activity was, however, first detected in 1952 through sero-epidemiological surveys in Nagpur district of Maharashtra and Chingleput district of Tamil Nadu. Japanese encephalitis as a disease was first reported in 1955 when cases of JE occurred in Vellore and Pondicherry in southern India. The virus was however, not recovered from man until 1958 when three isolations were made from the brain tissue of cases of encephalitis. This served to confirm JE as a case of encephalitis in India. Until early 1970s, the disease was reported only from southern India with periodic focal reports of its occurrence. A major outbreak resulting in 763 cases and 325 deaths [case-fatality rate (CFR)--42.6%] was reported from Bankura district of West Bengal in 1973. Subsequently, the disease spread to other states and caused a series of outbreaks in different parts of the country. In 1978, cases were reported from 21 states/UTs. Currently disease is reported from the states of Andhra Pradesh, Assam, Bihar, Goa, Haryana, Karnataka, Kerala, Mahrashtra, Manipur, Tamil Nadu, Uttar Pradesh, West Bengal and Nagaland. Till 2007 103389 AES/JE cases and 33729 deaths (CFR 32.62%) have been reported since 1978. Government of India launched vaccination campaign in highly endemic states of Assam, Karnataka, West Bengal and Uttar Pradesh in 2006 and in Andhra Pradesh, Bihar, Haryana, Maharashtra, Tamil Nadu in 2007 and 2008 respectively which has resulted in reduced incidence of JE in these states. PMID- 19552101 TI - Kala-azar elimination programme in India. AB - Kala-azar has been endemic in India for a long time. Phlebotomus argentipes, the vector became resurgent during 70s in four districts of Bihar and slowly spread to other parts of south Bihar and several districts of West Bengal. Kala-azar is a present endemic in 31 districs of Bihar, 4 districts of Jharkhand, 11 districts of West Bengal besides occurring is sporadic form in far districts of eastern UP. With enhanced central support to states from December, 2003 the case registration has improved. Elimination strategy lies in the followings: Case detection and treatment; interruption of transmission through vector control; advocacy, communication for behavioral impact and inter-sectoral convergence; capacity building; monitoring, supervision and evaluation and operational research. Moreover, inter-country co-ordination and assistance by Government of India are very important in elimination of kala-azar. PMID- 19552102 TI - Note on Kala-azar control programme in India. PMID- 19552103 TI - Elimination of lymphatic filariasis in India--a successful endeavour. AB - World Health Assembly resolution in 1997 for elimination of lymphatic filariasis (ELF) by 2020 made all the filaria endemic countries in the world to put efforts for its elimination by progressively reducing and ultimately interrupting the transmission of lymphatic filariasis. National Health Policy, 2002 has set the goal for elimination of lymphatic filariasis (ELF) in India by the year 2015. In pursuit to achieve this goal, the strategy of mass drug administration (MDA) with annual single dose of diethylcarbamazine citrate (DEC) tablets was introduced in 2004 in all the filaria endemic states of the country and 202 districts were covered. Intensive information education and communication (IEC) and awareness campaign were organised and more districts were included for MDA in subsequent years after 2004.Presently 250 districts are covered under MDA. The coverage of population during MDA against eligible population in the country has been reported 73.19% in 2004, 71.54% in 2005, 60.92% in 2006 and 81.41% in 2007. The microfilaria (mf) rate estimated in sentinel and random sites of each filaria endemic district revealed that in 2007, 177 districts are with less than 1% mf rate and 73 districts are with more than 1%. PMID- 19552104 TI - Emergence of drug resistance in India. AB - Chloroquine resistance in Plasmodium malaria is an emerging problem globally. In India resistance of Plasmodium falciparum to choloroquine, the cheapest and the most used drug was first reported in the year 1973 from Diphu of Karbi-Anglong district in Assam state. Systematic monitoring of drug resistance is being undertaken in the country from 1978 by the Directorate of National Vector Borne Disease Control Programme (NVBDCP) through its 13 Pf monitoring teams. The findings of these drug resistance studies has helped the programme for the revision of the drug policy and update it from time to time thereby facilitating appropriate measures for not only individual cases but also to contain and prevent further spread of resistant foci. This article summarises therapeutic efficacy studies conducted by the Pf monitoring teams in the country between 2001 and 2007 related to efficacy of chloroquine and other antimalarials drugs. As per the results available, the efficacy of chloroquine for treating uncomplicated falciparum at most of the study sites is much lower than the desired cut off levels of 10% (83% studies have shown treatment failure more than 10%). Total of 4273, 168 and 137 P. falciparum cases have been tested against chloroquine, sulphadoxine/pyrimethamine and ACT(AS+SP) combination. During the period under report, 85 new chloroquine resistant PHCs/foci from 64 districts were qualified warranting change of drug policy as per the national guidelines. These studies show that chloroquine resistance in P. falciparum is widespread in the country. To combat the drug resistant in malaria, the use of combination therapy ie, artesunate plus sulfadoxine/pyrimethamine has been recommended for treatment of all confirmed P. falciparum cases in all the qualified areas as per the criteria laid down in National Drug Policy on malaria. PMID- 19552105 TI - Stenting of non-malignant superior vena caval occlusion: a case report. AB - Endovascular stenting has been proposed as the primary treatment of choice in superior vena caval syndrome. Here a 34 year male had been diagnosed to have superior vena caval syndrome with pleural effusion (left side). CT scan showed sequential occlusion of superior vena cava lumen with left sided chronic pleural collection. Angiography showed superior vena caval occlusion. He was treated with antituberculous drugs for left sided pleural effusion. Superior vena caval syndrome was managed by deploying a stent which was postdilated by a peripheral balloon. PMID- 19552106 TI - Subcutaneous filariasis. PMID- 19552107 TI - Dynamics of the mammalian sperm head: modifications and maturation events from spermatogenesis to egg activation. AB - Mammalian spermatozoa have complex structures. The structure-function relationship of sperm has been studied from various viewpoints. Accumulated evidence has shown that the sperm components undergo sequential changes from the beginning of spermatogenesis to the time of fertilization/embryogenesis. Structural analyses have been performed using various new techniques of light and electron microscopy as well as immunohistochemistry and immunocytochemistry in combination with specific probes such as antibodies against sperm components. Recently developed gene-manipulation techniques have accelerated investigations on the events that govern the relationship between the structure and the molecular components of sperm. In addition, animal models with gene manipulations have been shown to exhibit various morphological and functional abnormalities that lead to infertility. In humans, male infertility is caused by a number of factors, such as the external environment, nutrient changes, genotoxins, and mutagens. These factors affect not only the development of germ cells during spermatogenesis but also the functions of mature sperm, ultimately impairing fertilization or embryogenesis. Typical phenotypes of impaired fertilization or embryogenesis are visible in conditions such as azoospermia, oligozoospermia, and teratozoospermia, which have been induced in model animals with gene deficiencies. Thus, comparative analyses of the phenotypes expressed by model animals carrying gene mutations and infertile human patients should be performed in relation to the normal (natural) fertilization process to clarify the etiology of infertility due to male factors. In this book, I discuss the events that occur in the normal sperm head and govern the structure-function relationship from the time of spermatogenesis to that of fertilization or egg activation. In this regard, I describe dynamic modifications and maturation events that occur in sperm-head components and compare the outcomes of these events with the outcomes of their failure. PMID- 19552108 TI - Identification and characterization of neural progenitor cells in the adult mammalian brain. AB - Adult neurogenesis has been questioned for many years. In the early 1900s, a dogma was established that denied new neuron formation in the adult brain. In the last century, however, new discoveries have demonstrated the real existence of proliferation in the adult brain, and in the last decade, these studies led to the identification of neural stem cells in mammals. Adult neural stem cells are undifferentiated cells that are present in the adult brain and are capable of dividing and differentiating into glia and new neurons. Newly formed neurons terminally differentiate into mature neurons in the olfactory bulb and the dentate gyrus of the hippocampus. Since then, a number of new research lines have emerged whose common objective is the phenotypical and molecular characterization of brain stem cells. As a result, new therapies are successfully being applied to animal models for certain neurodegenerative diseases or stroke. This work is being or will be extended to the adult human brain, and so it provides purpose and hope to all previous studies in this field. We are still far from clinical therapies because the mechanisms and functions of these cells are not completely understood, but we appear to be moving in the right direction. PMID- 19552109 TI - An agenda to damage. PMID- 19552110 TI - Frontline pressures after baby P. PMID- 19552111 TI - Controversial questions (part one): what is the right size for a health visiting caseload? AB - Questions asked by managers, commissioners and policy makers to find out what is, or should be, happening within health visiting services can seem immensely helpful in focusing the mind or clarifying key points. Alternatively, they may feel hostile and accusative, if their starting assumptions are alien to the everyday experience of health visitors. This paper is the first in a short series of three that draw on the experience of providing evidence to the Health Select Committee's 2008 inquiry into health inequalities. A formal process of seeking written evidence was followed up with specific questions in oral session, asked by committee members, trying to find out about how health visiting services relate to health inequalities. This line of questioning reflects concerns expressed elsewhere about the variability of health visiting services across the country and the lack of clear alignment to areas of deprivation, leading to calls for an increase in targeted services, instead of universal ones. This paper explores the notion of 'caseload', the distribution of services according to levels of deprivation and delivery of a universal or targeted health visiting service. PMID- 19552112 TI - Nothing about us without us: involving families in early support. AB - Early Support is the England-wide government programme to achieve child-centred and family-focused services for children aged nought to five years with a disability or complex need and their families. Integral to the ethos of Early Support is the understanding that every decision should be influenced and led by children and families. Families are expected to play a strategic role in the development and monitoring of policy and practice, and the service is expected to be proactive in seeking their views. Section 242(1B) of the NHS Act 2006 now places a duty to involve service users in planning the provision of services, changes to services and the operation of services. Yet for many frontline staff, user involvement and partnership working in this way presents real challenges. Using a framework based on principles of interpretive phenomenology, this paper documents efforts to ensure that the stories of families are heard and used to inform the development of proposals for Early Support in Bedfordshire and explores ways to overcoming barriers to meaningful user-involvement. PMID- 19552113 TI - Alcohol brief interventions: exploring perceptions and training needs. AB - One of the main thrusts of contemporary alcohol misuse policies is that early intervention can make a real difference to patterns of problem drinking, as long as healthcare professionals are given the right skills. However, healthcare professionals themselves suggest that they are often unsure of how to raise issues, and feel that they lack the skills or knowledge to do this effectively. This study investigates the perceptions and training needs of health visitors, school nurses, nursery nurses and practice nurses in relation to alcohol misuse primary prevention and the delivery of brief interventions in their day-to-day work. The findings indicate a variation in the need for training, which is reflected by the level of knowledge, skills and confidence of the different professional groups. This may help to facilitate the provision of needs-led alcohol training, and promote the effective delivery of support and brief interventions to individuals, families, schools and communities. PMID- 19552114 TI - Prescription to practise. PMID- 19552116 TI - Saving community services. PMID- 19552115 TI - New infant growth charts. PMID- 19552117 TI - Valuing health at work. PMID- 19552118 TI - Hospital design compromises care. PMID- 19552119 TI - Last month's prudence. PMID- 19552120 TI - A sad farewell to Sulu Fesili. PMID- 19552121 TI - Supporting Alzheimer's research. PMID- 19552122 TI - Over staffed, over paid and over here. PMID- 19552123 TI - How can nurses' flu vaccination rates be boosted? PMID- 19552124 TI - Encouraging nurses to get vaccinated. PMID- 19552125 TI - Death highlights poor ulcer care. PMID- 19552126 TI - Caring for those who care. PMID- 19552127 TI - Keeping staff and the organisation safe. PMID- 19552128 TI - Toxic mould syndrome--can the link be proved? PMID- 19552129 TI - Learning to handle patients safely. PMID- 19552130 TI - Take care when making statements. PMID- 19552131 TI - Employers' responsibilities are employees' complaints. PMID- 19552132 TI - Wharangi Ruamano. PMID- 19552133 TI - Surviving cancer. PMID- 19552134 TI - Identifying mistakes. PMID- 19552135 TI - We need more nurses. PMID- 19552136 TI - Disease update: professionitis. PMID- 19552137 TI - Breast self examination no longer recommended. PMID- 19552138 TI - New era dawns at Nursing Council. PMID- 19552139 TI - A passion for oncology nursing. PMID- 19552140 TI - The lonely road--a patient's view. PMID- 19552141 TI - Easing the transition to palliative care. PMID- 19552142 TI - Gardasil: holy grail or false idol? PMID- 19552144 TI - Making the case for gun law reform. PMID- 19552143 TI - Survey provides snapshot of nurses' attitudes to their work. PMID- 19552145 TI - Is your extended practice safe? PMID- 19552146 TI - Delegate or delegate? PMID- 19552147 TI - Developing the Maori nursing and midwifery workforce. PMID- 19552148 TI - [What kind of health care do we want for the 21st century?]. PMID- 19552149 TI - [Use and maintenance of a peripheral venous catheter for more than three days]. AB - The use of vascular access in a frequent procedure in the field of hospital practice. Nonetheless, vascular access is not bereft of complications, phlebitis being the most common. Following the Center for Disease Control and Prevention (CDC) recommendations, in our internal medicine service, changing a peripheral venous catheter is systematically carried out every 48-72 hours. The objective of this present study is to evaluate the need to change the vein accessed systematically every 48-72 hours, following the Center for Disease Control and Prevention (CDC) recommendations, in those patients who have a catheter inserted in a peripheral vein blocked by the administration of medication as part of the treatment of their pathology; or to evaluate the need to establish a new operational protocol since, according to other studies, its utility has not been demonstrated in a convincing manner. As a final conclusion, and while awaiting the results obtained in this study the authors believe it is possible to amplify the established periods for changing peripheral venous catheters while guaranteeing the security and comfort of patients having peripheral venous catheters inserted. PMID- 19552150 TI - [Care plan for the user of a Bricker-type urostomy]. PMID- 19552151 TI - [The Zero Bacteremia Project. Reduction of bacteremia caused by central venous catheters at ICUs in Spain]. PMID- 19552152 TI - [Nursing procedures and techniques: permanent pleural drainage devices for use after thoracic surgery]. AB - After various years of not having any innovations in the structure and manageability of pleural or thoracic drainage systems, at present time medical personnel are utilizing some known as permanent pleural drainage devices. These mechanisms are carried by patients who suffer from empyema or persistent pleural drainage, usually as a consequence of oncologic lung procedures. Current closed thoracic drainage systems are disposable and guarantee optimal, safe use by patients. To know the principal basic techniques for their correct utilization will allow medical personnel to maintain an efficient and effective drainage system as well as follow-up control of patients as their treatment evolves. In this article, the authors refer to the technical and the physiological aspects of this procedure so one fully understands how these devices function; they deal with the care necessary until these devices are finally removed. The authors describe nursing treatment and guides which provide recommendations focused on patient release from hospital care. PMID- 19552153 TI - [Mental health in contemporary women]. AB - The increase in illnesses and behavior disorders which medical personnel have observed among contemporary women during clinical practice in the outpatient services for Primary Health and Mental Health Care, basically anxiety and depression related disorders, is the reason why the author has written this article. This fact leads us to reflect on what is occurring. PMID- 19552154 TI - [Prevention of bed sores; results of a program in intensive pediatrics treatment]. AB - The difficulty to keep our patients' skin intact combined with few articles on this topic in pediatrics led the authors to carry out a study in their Pediatrics Intensive Care Unit "UCIP" at the Central University Hospital of Asturias, HUCA. The objective of this study consisted in determining if a decrease in the appearance of bed sores in children checked into their unit after a program whose purpose was to prevent bed sores had been applied occurred. This retrospective/prospective, comparative, observational study dealt with patients checked in their UCIP from September 2004 to September 2007; patients were broken into two groups, before and after applying a protocol and comparing the following variables: the number of patients checked in during each of the periods of this study; patients who had bed sores; average time kept in this ward for each group; pathologies patients who suffered from bed sores had when checked into this ward; the site and degree of these bed sores. Although there was an increase in the number of patients checked into this unit, the number of bed sores among children in this unit decreased 24.6% during the second period after the application of the protocol. Among the study observations noted there was a reduction in the severity of lesions patients manifested, as well as important differences as to where their lesions were located. Based on the results of the authors' study, one may conclude that the measures included in the bed sore prevention protocol, as well as the use of Mepentol hyperoxygenated fatty acids, have proven effective in their UCIP leading to a decrease in the number of patients which evidenced bed sores and the severity of these lesions was much less. PMID- 19552155 TI - [Oral anticoagulant treatment]. AB - Oral anticoagulant treatment is a preventive measure against thromboembolic disease; while this measure has very beneficial aspects, it is not exempt from risks since it has a few peculiar characteristics such as possible hemorrhages. The increasing number of patients who require anticoagulant medicine means that nurses may have to treat these patients in any forum where nurses work and therefore nurses need to know the traits this treatment measure entails. This article reviews the general characteristics of anticoagulant treatment and nurses' role in controlling this treatment. PMID- 19552156 TI - [Polyurethane foam dressing. Assessment of its therapeutic effects in the treatment of moderate- to highly-exudative chronic ulcers]. AB - The authors evaluate the behavior of Skinfoam Polyurethane Foam Dressings in the treatment of bed sores and vascular ulcers having a moderate to high exudation level by means of a multi-centric, prospective, open, observational study of a series of clinical cases. The length of this study was up to 20 dressing changes or until the patient was cured. During the first visit with patients, the personnel recorded patient data including an evaluation of the characteristics of patient's lesion; during visits to change dressings, the medical personnel recorded the evolution of the lesion and dressing changes; during the final visit, the medical personnel evaluated the final characteristics of the lesion as well as a global evaluation of the behavior of the dressing. This study included 185 patients who had an average age of 75.5. At the end of this study, 44.32% of these patients did not have any signs of an ulcer while among the other patients, the medical team noted an important decrease in the size of their lesions, with a considerable increase of tissue epithelization. The perilesional skin showed improvement in all aspects and erythema and eczema diminished in a statistically significant manner. The presence of exudation was null when the final dressing change occurred. Patients manifested a lack of pain before the end of this study 85.15% of the professionals indicated a global satisfaction level of good, very good or excellent while 100% of the patients stated they would use this dressing again. The authors conclude that the dressing under study is effective in the treatment of bed sores and high exudation vascular ulcers; even in cases involving compressive therapy, this dressing could be the one chosen for application. PMID- 19552158 TI - FTC's 'Red Flags' rule will affect most health care providers. PMID- 19552157 TI - Seize the EHR incentive. PMID- 19552159 TI - What to look for in e-prescribing advanced functions. PMID- 19552160 TI - Is your surgical practice missing revenue by not using modifier 24? PMID- 19552161 TI - The upper crust. PMID- 19552162 TI - PCs or thin clients? PMID- 19552163 TI - Making web visits work for your practice. PMID- 19552164 TI - Don't minimize EHR disruption, manage it. PMID- 19552165 TI - Economic stimulus package means landmark EHR incentives. PMID- 19552166 TI - Is CMS the IRS of medical practice? PMID- 19552171 TI - [Personnel's rights and obligations in elder abuse in institutions]. PMID- 19552172 TI - [Geriatric oncology, a new field of activity for the health networks?]. PMID- 19552173 TI - [Diagnosis disclosure oncology nurse. Interview by Tristan Cudennec]. PMID- 19552174 TI - [From competence to diploma, the VAE (validation of acquired experience) nursing aide in practice]. PMID- 19552175 TI - [Quality of life project and care project]. PMID- 19552176 TI - [Quality of life project or object of care, what alternatives for the very old?]. PMID- 19552177 TI - [Professional competences and a humanist approach at the heart of individualised quality of life projects]. PMID- 19552178 TI - [Quality of life project and care project in day care admission for Alzheimer's or similar diseases]. PMID- 19552179 TI - [Quality of life projects and personalized care projects in geriatric hospitals]. PMID- 19552180 TI - [Clothing care, a key element in the construction of a quality of life project]. PMID- 19552181 TI - [Thinking about geriatric care in the Breton language]. PMID- 19552182 TI - [External evaluation of a quality of life project in Ehpad using the quality indicator reference]. PMID- 19552183 TI - [Nurses, patients and families. 6/8--Dependency and physical, financial and material aides]. PMID- 19552184 TI - [3/6--Glaucoma]. PMID- 19552185 TI - Screening guidelines can vary based on risk. PMID- 19552187 TI - What has been your own experience with cancer survivorship? Oncology nurse brings unique perspective to caring for survivors. PMID- 19552186 TI - Life after cancer. AB - Many times we celebrate with our patients the last day of their cancer treatment. Often we may commemorate the occasion with a small party, a cake, or a token of some sort. We exchange hugs and well wishes, and, to an observer, it would appear that the last chemotherapy or radiation treatment is an event surrounded by happiness and relief. But for millions of cancer survivors, it's also a time of fear and uncertainty. PMID- 19552188 TI - Learn to recognize depression in patients with head and neck cancer. PMID- 19552189 TI - Health care and the environment: hospitals take steps to reduce waste. PMID- 19552191 TI - [Help, Vesale!]. PMID- 19552192 TI - [Organ trafficking: what reality?]. PMID- 19552190 TI - New study sheds light on angiogenesis inhibitors. PMID- 19552193 TI - [Multiple pulmonary abcesses and right side endocarditis]. PMID- 19552194 TI - [Immunologic thrombocytopenic purpura: a revolution of biological therapy?]. PMID- 19552195 TI - [The issue of stress]. PMID- 19552196 TI - [Herpes encephalitis]. PMID- 19552197 TI - [A clinical trap: acute and chronic psychogenic dyspnea]. AB - Psychogenic dyspnea may be part of two main clinical pictures. The first one is acute with dyspnea in the foreground of a panic disorder with or without agoraphobia. The second one is chronic, dyspnea being a background symptom of a generalized anxiety disorder. The problems related to the diagnosis and the implementation of psychological or pharmacological treatments are reviewed with reference to evidence-based medicine. PMID- 19552198 TI - [Adult hearing loss. From electronics to molecular biology]. PMID- 19552199 TI - [Pathophysiology of auditory and speech perception]. AB - Auditory perception or hearing can be defined as the interpretation of sensory evidence, produced by the ears in response to sound, in terms of the events that caused the sound. We do not hear a window but we may hear a window closing. We do not hear a dog but we may hear a dog barking. And we do not hear a person but we may hear a person talking. Hearing impairment can result in anxiety or stress in everyday life. Pure-tone hearing loss (or threshold shift) is a measure of hearing impairment. Aging and excessive noise are the main causes of hearing impairment. Speech perception is another concept. The difference with the former is best illustrated by the disabled individual declaring "I can hear that someone is talking to me, but I don't understand what she says". Being unable to understand easily and clearly significant others, especially in understanding speech in a noisy environment, can give rise to considerable psychosocial and professional consequences (disability). Presbycusis is the decline in hearing sensitivity caused by the aging process at different levels of the auditory system. However, it is difficult to isolate age effects from other contributors to age-related hearing loss such as noise damage, genetic susceptibility, inflammatory otologic disorders, and ototoxic agents. Therefore, presbycusis and age-related hearing loss are often used synonymously. In this report pathophysiology is mostly described with regard to presbycusis, and the main peripheral types of presbycusis (sensory or Corti organ-related, strial, and neural) are summarized. An original experimental model of strial presbycusis, based on chronic application of furosemide at the round window, is further described. Central presbycusis is mainly determined by degeneration secondary to peripheral impairment (concept of deafferentation). Central auditory changes typically affect speed of processing and result in poorer speech understanding in noise or with rapid or degraded speech. Last, age-related cognitive factors can be associated to peripheral hearing impairment and increase disability in speech understanding in noise. PMID- 19552200 TI - [Genetic deafness in adults]. PMID- 19552201 TI - [Prevention and management of acoustic traumas]. AB - Exposure to intense noises during a short time can be responsible for permanent damage of the inner ear, even after an isolated exposure. Amplified music (discos, concert rooms) is the first cause of acute acoustic traumas among the young people. Acute acoustic traumas need urgent management and treatment. Exposure to high noise levels during a long time can lead to a progressive and insidious impairment of the inner ear, the hearing loss appearing obvious only after several months or years. Noise-induced hearing loss is irreversible. The functional consequences of acoustic traumas can be socially disabling and can have an important psychological impact. The prevention is based on the information of the patients, notably of the youngest ones, and on the wearing of hearing protectors at the time of risky situations. PMID- 19552202 TI - [Adult deafness: hearing aid or hearing repair...]. AB - With the increase in lifespan, presbyacusis is a common situation. It starts with the difficulty of understanding in noisy conditions and may extend to withdrawal of social activity because we do not face the unintelligibility during diners and meetings. But, with difficulty, the hearing impaired confesses his deafness and hardly recognizes "his" presbyacusis. It will last approximately 10 years, 10 years with lost of profit from his entourage. The practitioner who meets more frequently this range of 60-65 years people should be able to detect this beginning deafness, or set the stage of presbyacusis. The obstruction of the ear canal by wax, that is not one, requires an active approach. These are the semiological elements we need to know for these 2 quite common situations. PMID- 19552203 TI - [Hearing loss in adults]. AB - The management of hearing loss in adults depends of etiology and its severity. It can be as simple as treating an external otitis, removing an impacted cerumen or a more complex one such as a surgery for otosclerosis. The hearing loss is managed mainly by new advances in hearing aids technology and implantable hearing devices which include BAHA, middle ear implant and cochlear implants. The research is focused on developing new molecules for intracochlear drug therapy to treat noise induced hearing loss, drug ototoxicity as well as hearing loss related to cochlear implant insertion trauma. Antioxidant molecules, molecules against apoptosis are at this time the most promising molecules than need further investigations. PMID- 19552204 TI - [Hearing loss among handicapped persons]. AB - Deafness appears to have a various impact on daily life, depending on the severity of the hearing loss. More often, mild and medium hearing loss are quickly rehabilited with hearing aids. Severe and profound hearing losses require a multidisciplinary evaluation, to determine the best treatment (speech therapy, cochlear implantation). Audiologic evaluation can be completed by handicap inventory specific for hearing loss, in order to appreciate the individual impact. Recently in France (2005) a law has demonstrated the determination of State's institutions to improve the daily life with handicap, and particularly in case of deafness. PMID- 19552205 TI - [Public assistance for the management of hearing loss]. AB - Financial management of adult deafness is still limited nowadays: the administrative assistance for purchasing hearing assistive devices, if not nonexistent, is often complex, difficult to implement and inconsistent. Therefore, not only patients but also recommending practitioners are often unaware of its existence. This article aims at providing below a non-exhaustive summary table, as simple as possible. PMID- 19552206 TI - [Leptospirosis]. AB - Leptospirosis is among the most important zoonotic disease worldwide. Annually, millions of human cases occur worldwide, with case fatality rates ranging as high as 20-25% in some regions. The pathogenicity of the Leptospira spp is depending on the responsible serovar. Rodents are the most important primary hosts but a wide range of other mammals are also able to carry and transmit the disease as secondary hosts. Humans become infected directly through infected animal contact or indirectly with water, food, or soil containing urine from these infected animals. The main route of infection is the skin through skin abrasions, more rarely through mucosa contact and exceptionally by respiratory or digestive route. The diagnosis should be evoked on a detailed personal history evoking epidemiological context with consistent clinical and biological results. A lot of infected persons may have no symptoms at all showing a minor illness. In most cases symptoms are flu-like with fever, headache and intensive myalgia associated with hepatic cytolysis, proteinuria and thrombocytopenia. In France, the most common clinical presentation combines liver involvement and jaundice with kidney involvement. However, other sites can be involved (brain, heart and lung). The diagnosis is often obtained several weeks later by serology. Treatment for leptospirosis is based on an early antibiotic therapy with amoxicillin, or tetracycline in case of allergy, but it remains controversial whether antimicrobials produce a beneficial effect in mild human leptospirosis. For at risk occupations, preventive treatment recommends eviction or limitation of exposure, and vaccination. PMID- 19552207 TI - [Managing conflicts of interest]. PMID- 19552208 TI - [Aspects of observational epidemiologic studies: case series, cross-sectional studies, environmental studies, case-control studies]. PMID- 19552209 TI - [Palpitations]. PMID- 19552210 TI - [Sanitary risks linked to water and food. Food poisoning]. PMID- 19552211 TI - [Epilepsy in the child]. PMID- 19552212 TI - [Multiple sclerosis]. PMID- 19552213 TI - [Documentary research and self-instruction]. PMID- 19552214 TI - [Different methods of psychotherapy]. PMID- 19552215 TI - [Indications and strategies of diagnostic imaging modalities]. PMID- 19552216 TI - [Movement disorders]. PMID- 19552217 TI - [First French observation of cystic fibrosis of the pancreas (mucoviscidosis)]. PMID- 19552218 TI - A response to "the hassle factor". PMID- 19552219 TI - Ask and act: Delaware physicians demonstrate the effectiveness of the American Academy of Family Physicians' initiative to promote tobacco cessation counseling. AB - The Ask and Act program of the American Academy of Family Physicians promotes family physician tobacco cessation counseling. Smoking and cessation counseling rates were obtained by cross-sectional analysis of electronic medical record data before and after initiation of the Ask and Act program in Delaware. Before the intervention, 24% of the patient population sampled were smokers and 70% of smokers had been counseled to quit smoking. After the intervention, 17% of the patient population sampled were smokers and 98% of smokers had been counseled. Our observations suggest that the Ask and Act program increases counseling, thereby increasing smoking cessation. PMID- 19552220 TI - The true cost and benefit of medication reduction. PMID- 19552221 TI - More on access. PMID- 19552222 TI - The role of pediatric dentistry in multidisciplinary cleft palate teams at advanced pediatric dental residency programs. AB - PURPOSE: The purpose of this study was to characterize the participation of pediatric dentistry in multidisciplinary cleft palate teams (CPTs) at advanced pediatric dental residency programs. METHODS: A survey was sent to the directors of advanced pediatric dentistry programs across the United States. RESULTS: Of the 60 (90%) surveys returned, 18% of the programs were university-based, 40% hospital-based, and 42% combined programs. Overall, 92% of the programs reported pediatric dentistry's participation in CPTs. Orthodontics, plastic surgery, oral surgery, otolaryngology, and speech therapy, are represented on at least 75% of the CPTs. Nursing and psychology are represented in less than 50% of the CPTs. A higher percentage of combined programs reported providing interceptive orthodontics, while more hospital-based programs reported providing presurgical infant orthopedic appliances (PIOAs). Of the 47% of the programs that reported use of POIA, 64% reported using removable appliances. Seventy-five percent of the programs reported that there has been no change, 22% reported an increase, and 3% reported a decrease in the CPT participation level in the post 5 years. CONCLUSIONS: This study highlights the role of pediatric dentistry as a part of cleft palate team. This role extends from preventive and restorative to infant orthopedics. PMID- 19552224 TI - Infant oral health examinations: pediatric dentists' professional behavior and attitudes. AB - PURPOSE: The purposes of this study were to: (1) determine pediatric dentists' behaviors and attitudes concerning infant oral health examinations (IOHEs); and (2) explore how respondents who do or do not perform IOHEs differ in their behavior and attitudes concerning IOHEs. METHODS: Survey data were collected from 2157 members of the AAPD (response rate=49%). Most respondents were male (67%) and practiced in solo practices (45%). RESULTS: While 84% of the respondents agreed with the AAPD policy concerning IOHEs and reported that they perform IOHEs, only 53% indicated that they see infants 12 months old or less. The majority of respondents engaged in oral hygiene education (86%) and performed soft and hard tissue exams (81% and 75%, respectively). Fewer practitioners assessed family dynamics (33%), or talked about injury prevention (53%). The respondents had positive attitudes concerning IOHEs. These attitudes were correlated with the number of IOHEs conducted. CONCLUSIONS: The finding that only 53% of the respondents see 1-year-old children or younger shows that efforts need to continue to increase the percentage of dentists who offer IOHEs. Most respondents held rather positive attitudes toward IOHEs. They differed in the amount of time they schedule for these exams and the issues they address. PMID- 19552225 TI - Preventive resin restorations: practice and billing patterns of pediatric dentists. AB - PURPOSE: The purpose of this study was to determine practice patterns of pediatric dentists for preventive resin restorations (PRRs) and if they believe a code should be added to the American Dental Association's current dental terminology (CDT) for the PRR. METHODS: A 16-question survey sent to 475 pediatric dentists randomly selected from the American Academy of Pediatric Dentistry database, addressed demographics, treatment planning, techniques in preparation and restoration, billing practices, and perceptions about the need for a CDT code for PRRs. RESULTS: Two hundred thirty-eight (50%) surveys were returned, revealing that 72% of respondents perform PRRs and 64% feel that a PRR code should be added to the CDT Fifty-two percent believe not having a CDT code could cause dentists to perform more invasive dentistry to comply with billing requirements that Class I restorations be in dentin. PRRs are commonly treatment planned for deep pits and fissures with questionable decoy not entering dentin. Up to 50% of respondents could be erroneously billing for PRRs. CONCLUSIONS: Most pediatric dentists perform preventive resin restorations in their office and believe that a code for the procedure needs to be added to the current dental terminology. PMID- 19552223 TI - Maternal transmission of mutans Streptococci in severe-early childhood caries. AB - PURPOSE: This study utilized multiple genetic analyses to detect evidence of maternal MS acquisition in children with S-ECC. METHODS: Twenty-seven mother/child pairs were selected from children with 5-ECC preceding dental rehabilitation under general anesthesia. Plaque samples were collected from the mother, child, and the child's carious lesion. Arbitrarily primed-polymerase chain reaction (AP-PCR) genotyped 6-8 MS isolates from each plaque sample, and unique genotypes were identified. Representative MS isolates with unique genotypes were characterized by pulsed-field gel electrophoresis (PFGE). Cluster analysis using the Dice band-based similarity coefficient was used to generate dendrograms from gel bonding patterns. A Dice coefficient >70% indicated similarity or match among PFGE genotypes. RESULTS: In 26% (7/27) of mother/child pairs, all of the child's isolates matched the mother. In 15% (4/27), some of the child's genotypes matched the mother, and in 59% (16/27), no isolates matched the mother. Maternal transmission was a mode of MS acquisition in 41% (11/27) of mother/child pairs, while acquisition from non-maternal sources occurred in 74% (20/27). CONCLUSIONS: MS genotypes that did not match maternal strains were identified in the majority of children (74%) within this S-ECC population. Evidence of maternal transmission was detected in 41% of mother/child pairs. PMID- 19552226 TI - Body mass index of children with severe early childhood caries. AB - PURPOSE: The purpose of this retrospective case study was to describe the body mass index (BMI) of children with severe early childhood caries (5-ECC) receiving dental rehabilitation under general anesthesia. METHODS: Demographics, BMI percentile, decoyed, missing, or filled teeth (dmft), and the number of pulp involved teeth were analyzed for 293 healthy 2- to 5-year-olds (mean = 47.2 months). Weight groups were assigned using current Centers for Disease Control (CDC) BMI-for-age and gender definitions. Descriptive statistics were calculated and multivariate analysis used to evaluate BMI's association with oral health measures. BMI distribution of the subjects was compared graphically and with the use of confidence intervals to a reference population with similar demographics. RESULTS: The distribution of subjects' BMI percentiles was: underweight=11l%; normal weight=67%; at risk for overweight=9%; and overweight=11%. The mean dmft was 11.8; BMI percentile did not correlate with dmft or the number of pulp involved teeth. Significantly, more children in the sample were underweight than in the reference population (11% vs. 5%). CONCLUSION: In this sample of S-ECC children, the BMI percentile was not correlated with dmft or the number of pulp involved teeth, even After adjusting for confounding factors. Thirty-two percent had unhealthy weights, as currently defined by the Centers for Disease Control and Prevention. PMID- 19552227 TI - Effect of sensory adaptation on anxiety of children with developmental disabilities: a new approach. AB - PURPOSE: The aim of this study was to evaluate the effect of a sensory-adapted dental environment (SADE) on anxiety, relaxation, and cooperation of children with developmental disabilities (CDDs). Pharmacological treatment has been widely used to reduce anxiety, but nonpharmacological methods may be similarly effective. The standardized clinical situation chosen was a dental hygiene cleaning. METHODS: A SADE was structured. Sixteen CDDs participated in an open cross-over intervention trial measuring behavioral and psychophysiological variables. RESULTS: There was a substantial increase in relaxation and cooperation in the SADE as opposed to the regular dental environment (RDE). This was reflected by: mean duration of anxious behaviors (SADE = 9.04 minutes vs. RDE = 23.44 minutes; P < .01); mean magnitude of anxious behaviors (SADE = 8.49 vs. RDE = 15.50; P < .01); cooperation levels (SADE = 331 vs. RDE = 1.94; P < .01); mean electrodermal activity (EDA; SADE = 1230 vs. RDE = 446; P < .001); and difference in degree of relaxation by EDA (SADE=2014 vs. RDE=763; P < .004). CONCLUSIONS: The findings indicate the potential importance of considering the sensory-adapted environment as a preferable dental environment for this population. PMID- 19552228 TI - Dental caries and its association with sociodemographics, erosion, and diet in schoolchildren from southeast Brazil. AB - PURPOSES: The purposes of this study were to determine the prevalence of caries in a sample of Brazilian schoolchildren, test associotions between caries, sociodemographics, and erosion, and investigate potential dietary factors common to both conditions. METHODS: Brazilian schoolchildren (13-14 years old) completed a questionnaire to provide dietary information and underwent dental examination. Sociodemographic characteristics were collected using a questionnaire completed by parents/guardians. Caries was assessed using decayed, missing, filled teeth/ surfaces (DMFT/DMFS) indices. Erosion was assessed using a previously validated index. RESULTS: Of 458 schoolchildren, 78% had caries experience. A statistically significantly lower prevalence of caries was observed in children from a higher economic class and whose parents had higher educational levels. Thirty-five percent of children with caries also had erosion, while 32% with a DMFT of 0 had erosion (P = .72). The frequency of consumption of drinks and foods was not statistically significantly associated with caries. The intake of sugared carbonated drinks was statistically significantly associated with erosion (P = .01). The mothers' educational level was the only variable independently associated with caries experience (P = .04). CONCLUSIONS: Dental caries was associated with sociodemographics, but not with erosion. There were no dietary factors identified in common for caries and erosion. PMID- 19552229 TI - Pediatric conditions associated with compromised airway: part I--congenital. AB - Epidemiological studies have confirmed that respiratory complications are collectively the leading cause of pediatric anesthetic-related morbidity and mortality Perioperative pulmonary complications associated with sedation and/or general anesthesia include obstruction, atelectosis, aspiration, pneumonia, bronchitis, laryngospasm, bronchospasm, hypoxemia and respiratory failure. This review discusses congenital conditions characterized by potentially diff cult airway management, and proposes treatment considerations to prevent adverse events in these patients. The importance of patient evaluation prior to sedation and general anesthesia is emphasized. Knowledge of normal and abnormal head and neck anatomy and its influence on maintaining airway potency during routine and emergency procedures is necessary in providing safe and high-quality care for young and adolescent patients. PMID- 19552230 TI - Lower lip mucocele treated with an erbium laser. AB - Mucoceles are benign lesions of the minor salivary glands that are common in children. The most frequent localizations of these lesions include the lower lip and the cheek mucosa. Such mucoceles are caused by traumas, the rubbing of orthodontic devices, or biting habits. The purpose of this article was to describe the case of a 4-mm extravasation mucocele located on the lower lip of a 9-year-old girl. This mucocele was removed with an erbium loser after perilesional infiltration with 12 mg of 2% lidocaine and epinephrine 1:100,000. The histopathological report confirmed the presurgical diagnosis. The wound healed excellently and rapidly without sutures. No relapse was observed a year ofter the surgery. Lasers apply modern technology and are useful for soft tissue surgery in pediatric dentistry, as operations are rapid and wounds heal well without sutures. PMID- 19552231 TI - Repositioning of a completely intruded permanent incisor with surgical extrusion: a 4-year follow-up case. AB - Intrusive luxation is a serious dental injury in the permanent dentition that has a poor prognosis. There is a lock of consensus concerning the management of intruded permanent teeth because of a small number of published studies and the injury's complicated nature. The purpose of this report was to present the successful outcome of a completely intruded permanent maxillary right central incisor with a concomitant uncomplicated enamel fracture in a 9-year-old boy. The intruded tooth was repositioned via surgical extrusion and splinted with a semirigid splint for three weeks. Clinical and radiographic examination 48 months often the surgical extrusion revealed no signs of progressive root resorption, marginal bone loss, or periopical disease. Also the percussion sound was normal and the tooth was functionally acceptable and oesthetically pleasing. PMID- 19552232 TI - Xylitol: effects on the acquisition of cariogenic species in infants. AB - PURPOSE: The purpose of this study was to examine the effects of xylitol gum (XG) on the acquisition pattern of 39 bacterial species, including mutans streptococci (MS), in infants. METHODS: Ninety-seven mothers (MS counts > 10(5) CFU/ml) were randomly divided into 4 groups and received: (1) XG (4.2 gm/day); (2) XG (6 months after baseline exams); (3) sorbitol gum (4.2 gm/day); or (4) no gum. Groups 1 and 3 chewed gum 3 times a day for 9 months. Microbiota of plaque and saliva samples from the mother-child pairs were analyzed by culturing and via checkerboard DNA-DNA hybridization. RESULTS: MS was isolated from 33% of the predentate infant (< or =5 months old) baseline saliva samples and from 41% of the saliva and 65% of the plaque samples at the final visit. At baseline, positive responses to "mother's checking of baby's food temperature using baby's spoon" and "starting a bottle after stopping breast-feeding" were significant predictors (P = .009 and P < .001, respectively) of infant's total streptococci counts. At the final visit (9 months later), there were no significant differences between treatment groups for infants' 39 microbial plaque species, including MS. CONCLUSIONS: Maternal use of xylitol gum did not result in statistically significant differences in the microbial plaque composition of 9- to 14-month-old infants. PMID- 19552234 TI - Problems with minimal-risk research oversight: a threat to academic freedom? PMID- 19552233 TI - An intervention to improve cancer patients' understanding of early-phase clinical trials. PMID- 19552236 TI - Reassessing "patient knowledge". PMID- 19552235 TI - Data-sharing dilemmas: allowing pharmaceutical company access to research data. PMID- 19552237 TI - Planning for a different future. PMID- 19552238 TI - How are you doing in the new economy? PMID- 19552239 TI - The critical role of physicians in fund development. PMID- 19552240 TI - Financial analysts' changing view of health care philanthropy: how does your program rate? PMID- 19552241 TI - Thoughts on being a successor. PMID- 19552242 TI - HIPAA, Sarbanes-Oxley, IRS, USPS and the states: juggling complicated U.S. regulations. PMID- 19552243 TI - Controversial role prompts action in Scotland. PMID- 19552244 TI - Confidentiality and children with HIV. AB - AIM: This study explored how healthcare professionals ensure confidentiality for children with HIV, whether these methods comply with the Data Protection Act and the Caldicott principles for confidentiality and whether the methods used enable healthcare professionals to manage disclosure of HIV status outside of the health service. METHOD: Nineteen healthcare staff participated in semi-structured interviews that were tape recorded. Data were analysed using the constant comparative method. FINDINGS: The measures that were described appeared to be consistent with data protection principles. A minority suggested that the correct processes were not always adhered to. A prominent theme was uncertainty about disclosure, particularly in relation to whether schools should be informed about the status of children with HIV. Effective communication among healthcare professionals appeared to be a significant factor in minimising the risk of breaches in confidentiality. CONCLUSION: Families can be reassured that confidentiality is being maintained within the organisation where this study was conducted. However the risk of inappropriate disclosure remains high, indicating that staff should not be complacent about this area of their practice. PMID- 19552246 TI - Diarrhoea and vomiting. PMID- 19552245 TI - Central venous access devices: review of practice. AB - Management of central venous access devices is a complex nursing activity with the potential for serious complications. There are variations in the care and maintenance of these devices, between and within, hospitals. Building on previous literature reviews and audits, a postal survey was undertaken to inform review of practice guidelines. METHOD: Questionnaires were sent to 22 national, tertiary paediatric surgical centres and neonatal units where neonatal surgery was undertaken. Respondents were asked to provide information on: types of access devices and dressings used, cleansing agents, solutions to flush the device and sizes of syringes used to access the device. RESULTS: Broviac catheters were the most common. There was considerable variation in practice with a range of dressings, cleansing agents, flush solutions and syringe sizes. CONCLUSIONS: Literature reviews, practice audits and the postal survey provide good evidenc for recommendations on dressing type, cleansing agent and syringe size. Selection of flush solution and amount to be used depend on manufacturer advice. Implementation of these practice guidelines can be supported by placing printed instructions at the bedside of each patient with a central venous access device. PMID- 19552247 TI - Decision making in nursing practice: a case review. AB - Clinical decisions and clinical judgements are important elements of nursing practice because they have a direct effect on patient care. A four-stage information processing framework was used to critique a decision to carry out chest examination and auscultation during the routine assessment of a child's asthma status. The use of this process demonstrates that nurses apply a combination of analytical and intuitive strategies when making decisions. Nurses who manage the care of patients in nurse-led clinics need to acquire advanced assessment skills to support their clinical decision making. PMID- 19552248 TI - Mental health of children and young people in hospital. AB - It is estimated that one in ten children and young people have mental health problems such as emotional or conduct disorders. Acute and chronic physical illnesses also have consequences for mental health and well being. In acute settings, physical health problems are well addressed but provision for mental health needs generally falls short of the holistic care that is necessary for optimum health. The Common Assessment Framework supports early identification of children's additional needs and provides for timely and co-ordinated support to meet those needs. However, it is not yet widely implemented in general inpatient settings. Staff need to work closely with child and adolescent mental health staff, addressing the challenges of information sharing and interagency working, if they are to provide more holistic care and support. PMID- 19552249 TI - Involving children in research. AB - Children and young people should be involved in the research process when the findings could affect them. It is important to use research tools that will facilitate their engagement as they can offer valuable contributions that generate a knowledge base that is a unique reflection of their needs. Visual methods such as drawing, collage and photography can help them to express emotions and opinions. Reference groups of children and young people can inform every stage of the research process, ensuring appropriateness of information materials, consent forms and methods. There are challenges associated with the involvement of children and young people in the research process, but with careful planning, these are surmountable. PMID- 19552250 TI - Managing stomal bleeds. PMID- 19552251 TI - Body image in adolescents: insights and implications. AB - This article explores some of the key processes associated with the development of a normal body image in adolescence and considers the impact of illness on the same. Nurses are assisted to explore the ways in which their insights into body image change might assist adolescents to cope with illness, injury or disability in a more constructive way. PMID- 19552252 TI - Imperfect blend. PMID- 19552253 TI - More on the number of embryos to transfer. PMID- 19552254 TI - Brave new field. PMID- 19552255 TI - A different view of depression. PMID- 19552256 TI - Complementary coursework. PMID- 19552257 TI - Responsibility is key to health care reform. PMID- 19552258 TI - Lone ranger. PMID- 19552259 TI - From the margins to the mainstream. PMID- 19552260 TI - Alternate route. PMID- 19552261 TI - A pharmaceutical economic stimulus plan. PMID- 19552262 TI - A primer on acupuncture. AB - Although widely practiced in China for 2,000 years, acupuncture was introduced in the United States 40 years ago, and it has only gained acceptance by the medical establishment here in recent years. Now, a growing body of evidence supports its efficacy for a number of conditions, and it is being routinely recommended for them. This article provides an overview of acupuncture and reviews some of the evidence showing its effectiveness for treating various conditions. PMID- 19552263 TI - Integrative holistic medicine in Minnesota. AB - Minnesota has played a leading role in the integrative holistic medicine movement in the United States for more than 2 decades. This article defines integrative holistic medicine and describes how it is practiced. It also discusses the reasons why institutions and providers here and elsewhere in the country have embraced this approach to patient care. PMID- 19552265 TI - The challenge of delivery of care. PMID- 19552264 TI - Mind-body medicine: a model of the comparative clinical impact of the acute stress and relaxation responses. AB - Although the physiological and biochemical changes that occur during the acute stress response have been well-characterized, the contrasting changes that underlie the relaxation response evoked by various mind-body techniques are less understood. To help guide future mind-body research, we present a conceptual model that integrates patterns of change at the physiological and molecular levels. In addition, we point to future research opportunities and discuss how repeated elicitation of these responses could influence the health of patients. PMID- 19552266 TI - Ke Ola Mamo: Oahu's Native Hawaiian Health Care System. PMID- 19552267 TI - Mobile Van clinic: helping to serve Maui's uninsured. PMID- 19552268 TI - Lutheran Medical Center's dental residencies--a history of breaking boundaries. PMID- 19552269 TI - The future of healthcare. Observations from a physician/legislator. PMID- 19552270 TI - An introduction to Hawai'i's community centers. PMID- 19552271 TI - The truth about attrition. PMID- 19552272 TI - Will measuring the quality of nursing care improve patients' experience? PMID- 19552273 TI - Time in motion. AB - With junior doctors' hours due to be reduced from August, nurses may find changes in their working lives. PMID- 19552274 TI - What the directive will mean. AB - The European Working Time Directive (EWTD) became law in the U.K. more than ten years ago, but some nurses still choose to work long hours. PMID- 19552275 TI - Home-made health. AB - Research into parents' use of home remedies for their children produced some startling results. Yet few healthcare professionals realise the extent of their use. PMID- 19552276 TI - Fitness regime. AB - A nurse-led programme to improve people vascular health is the aim of MyAction. PMID- 19552277 TI - No more tears. PMID- 19552278 TI - Working in a youth offending team: the learning disability nurse's role. AB - This article highlights the importance of learning disability nurses' skills to youth offending teams. Through the use of a case study, it explains why this role is making a difference in one location in the U.K. and how it could and should make a difference in other youth offending teams. PMID- 19552279 TI - Skin disinfection and its efficacy before administering injections. AB - The need to disinfect a patient's skin before subcutaneous or intramuscular injection is a much debated practice. Guidance on this issue varies between NHS organisations that provide primary and secondary care. However, with patients being increasingly concerned with healthcare-associated infections, a general consensus needs to be reached whereby this practice is either rejected or made mandatory. PMID- 19552280 TI - Interpretation and evaluation of pulmonary function tests. AB - Evaluation of pulmonary function is essential for the diagnosis and management of respiratory disease and other conditions. Various tests are available. Some, such as spirometry and pulse oximetry, are widely used in a variety of healthcare settings; others, such as those that measure diffusing capacity and static lung volumes, are performed in specialist pulmonary function laboratories. This article examines some of the pulmonary function tests available. PMID- 19552281 TI - Immune system. PMID- 19552282 TI - Lost in the hubbub. PMID- 19552283 TI - Sketching out a new career. Interview by Frances Pickersgill. PMID- 19552284 TI - Measures to stop the rot. PMID- 19552285 TI - Eradicating central venous catheter related infection. PMID- 19552286 TI - Unravelling Menieres disease: an approach to diagnosis & management. PMID- 19552287 TI - Deliberate Self-Harm (DSH): a follow-up study of Irish children. AB - This study aimed to look at rates of repetition in children presenting with Deliberate Self-Harm (DSH) to a paediatric A&E department. Children presenting with DSH to a paediatric A&E between 2000 and 2005 were invited to participate in the study. Telephone interviews collected information on demographic details and mental health functioning, including repetition of DSH. Data was obtained from 39 parents and 10 children (31 girls and 8 boys, mean age 15) 1 in 5 females (20%) had made a repeat attempt of DSH and 1 in 10 (10%) had repeated more than once. No males repeated self-harm. On going parental concern rather than clinician risk assessment at index episode predicted subsequent DSH. Given the poor predictive value of clinician risk assessment, all attempts of DSH must be taken seriously, especially those associated with ongoing parental concern. PMID- 19552288 TI - Usage of emergency contraception between medical related and non-medical related students. AB - Teenagers and young adultshave the most risk of unplanned pregnancy, due to lack of awareness to see a family planning provider after unprotected sexual intercourse. In addition, nearly one in five physicians is reluctant to provide information regarding Emergency Contraception (EC) to women and this may contribute to their lack of awareness. This study was conducted to assess the knowledge, attitudes and practices regarding the use of EC between medical related students compared to non-medical related students. Data collection was done using questionnaires distributed among students in University College Cork (UCC). 93% of medically related students were aware of EC compared to only 73.5% of non-medically related students. Medical related students also were more aware about the mechanism of action and detailed knowledge of EC compared to the non medical students. This study has proven that medically related students have more detailed knowledge regarding EC compared to non-medical related students. However, there was no significant difference noted regarding the attitude and practice between the two groups. PMID- 19552289 TI - Occupational injuries in foreign-national workers presenting to St James's Hospital Plastic Surgery service. AB - This study was undertaken to investigate our impression that migrant foreign national workers were more at risk of sustaining work place injuries requiring referral to our Plastic Surgery service than their indigenous Irish counterparts. Data were collected prospectively from August 2006 to February 2007 on all work related injuries presenting to the Plastic Surgery service in St James's Hospital, Dublin. 201 work-related injuries were recorded during the six month study period. 40% (n = 81) of the study group were foreign-national workers. Foreign-national workers account for only nine percent of the total Irish workforce. 31% (n = 25) of the study group required a translator. Over half (55%) of all the foreign-national workers in the current study had been in their present job for less than six months at the time of injury compared to only nine percent of Irish workers. This study highlights that foreign-national workers in Ireland are at a disproportionately high risk of occupational injury when compared to their Irish colleagues and emphasises the need for targeted occupational health and safety measures in this vulnerable group. PMID- 19552290 TI - Irish neonatal mortality statistics for 2004 and over the past 17 years: how do we compare internationally? AB - In the past 17 years neonatal mortality survey has provided important data on the trends in deaths of all live born infants born in Southern Ireland who are greater than 500 g birth weight and who die within the first 28 days of life. The aims of this study were to report neonatal mortality data for Southern Ireland for 2004, to examine trends in neonatal mortality over the past 17 years and compare Irish Neonatal Mortality rates to other countries around the world. The neonatal mortality rate for 2004 was 2.9/1000 with a corrected NMR of 1.9/1000. The response rate to the survey was 100%. Prematurity is now the leading cause of neonatal mortality representing a change from previous years. Deaths related to asphyxia have remained largely unchanged. When compared to international figures Ireland compares favourably to other countries around the world. PMID- 19552291 TI - EUROASPIRE (European Action on Secondary Prevention through Intervention to Reduce Events) III--a comparison of Irish and European results. AB - The EUROASPIRE III audit was a Europe-wide study which took place in 2006/2007. The objective was to examine the control of risk factors in subjects with established cardiovascular disease. Here, we compare the Irish results to those of the other 21 European countries which participated. Control of blood cholesterol was significantly better in Irish participants, with 73% below the target of 4.5 mmol/l. Blood pressure control was less satisfactory in both Irish and European individuals, with an average of 52% of Irish participants not achieving blood pressure targets. Medication usage was high throughout, particularly anti-platelet agents, beta-blockers and, especially in Ireland, statins. Obesity figures were particularly high in Ireland and throughout Europe, with 82% Irish men and women either overweight or obese. Smoking figures in Irish women were also of concern, with 24% continuing to smoke. Cardiac rehabilitation attendance was particularly high in Ireland, with 68% attending; substantially higher than the European figure of 34%. In common with the rest of Europe, current control of body weight and blood pressure in Ireland is unsatisfactory and in need of increased consideration on the part of both patients and healthcare professionals. PMID- 19552292 TI - Obesity in Ireland in 2008: what radiological equipment is available to image the obese patient? AB - Obesity is a global epidemic, responsible for 2000 premature deaths in Ireland each year. The extent of this epidemic was quantified by the National Taskforce on Obesity (IOTF), whose report, published in 2005, found that 39% of adults in Ireland were overweight and 18% obese with obesity in adults predicted to increase by 1% per year. In light of the clear evidence that we, as a nation, are quite literally expanding, how well equipped are Irish hospitals and, in particular, radiology departments, to deal with patients of increasing size and weight? The purpose of this study was to quantify the weight limits and girth restrictions of the radiology equipment, in particular CT, MRI and fluoroscopy, in hospitals, both public and private, in Ireland in an attempt to answer this question. PMID- 19552293 TI - Preoperative fasting time in children. AB - The aim of preoperative fasting is to prevent regurgitation and pulmonary aspiration while limiting potential problems of thirst, dehydration and hypoglycaemia. The American Society of Anaesthesiologists (ASA) has suggested guidelines for preoperative fasting for children undergoing elective surgery. We did a postal survey to determine the current practice regarding these guidelines amongst all specialist registrars in anaesthesia in Ireland. A questionnaire was sent to all specialist registrars in anaesthesia (90 in total), 60 (67%) were returned and analysed. The question asked was how long children should be kept fasting before elective surgery. The results of our survey suggest that most of the respondents are following the ASA guidelines for clear fluids and solids however there were differing opinion regarding the duration of fasting for formula milk and breast milk. In conclusion, we would recommend greater awareness and collaboration between anaesthetists, nurses and surgeons to ensure that fasting instructions are consistent with the ASA guidelines and that patient and their parents understand these directives as well. PMID- 19552294 TI - Isolated pancreatic metastases from a bronchogenic small cell carcinoma. AB - We describe the case of a 60 year old female smoker who presented with a three month history of weight loss (14 Kg), generalized abdominal discomfort and malaise. Chest radiography demonstrated a mass projected inferior to the hilum of the right lung. Computed Tomography of thorax confirmed a lobulated lesion in the right infrahilar region and subsequent staging abdominal CT demonstrated a low density lesion in the neck of the pancreas. Percutaneous Ultrasound guided pancreatic biopsy was performed, histology of which demonstrated pancreatic tissue containing a highly necrotic small cell undifferentiated carcinoma consistent with metastatic small cell carcinoma of the bronchus. PMID- 19552295 TI - Up front about frontal headaches and sinusitis. AB - In clinical practice frontal headaches are common however are frequently incorrectly attributed to rhinosinusitis. The misdiagnosis of frontal headaches and/or facial pain has been compounded by increased accessibility and over reliance on sinus CT imaging. We reviewed the presentation and clinical findings of 3 patients in our unit with frontal headaches misattributed to rhinosinusitis. We also reviewed the literature regarding the diagnostic criterias for rhinosinusitis and the role/limitations of CT imaging in the diagnosis of paranasal sinus disease. All 3 patients had isolated frontal headaches in association with normal nasal examination. They had also undergone CT imaging with isolated frontal sinus opacification evident in 2 cases and frontal and maxillary opacification in the third. All failed medical therapy and when CT imaging was repeated, it was found to be normal. The combination of facial pain/headache alone and sinus opacification on CT imaging do not meet the criteria for either acute rhinosinusitis or chronic rhinosinusitis. Other symptomotolgy, CT and endoscopic findings need to be considered to prevent the incorrect diagnosis of rhinosinusitis and unnecessary surgical intervention. For those cases where the clinical presentation does not support a diagnosis of rhinosinusitis, repeat CT imaging and a neurology consultation may help in clarifying the diagnosis and deciding which patients really do need surgery. PMID- 19552296 TI - Transfer of patients following non-elective PCI is safe after four hours of observation. PMID- 19552297 TI - Diagnosing Takayasu's arteritis with modern day radiology. PMID- 19552298 TI - The uses of mercury equipment and products in Irish healthcare. PMID- 19552299 TI - First Irish pregnancies after IVF with gestational carrier. PMID- 19552300 TI - Timing and modality of neuroimaging in acute stroke: an Irish perspective. PMID- 19552301 TI - Introduction to neuroeconomics. AB - Neuroeconomics has quickly attracted the attention of economists, psychologists, and neuroscientists. These scholars have joined to combine experimental methods and mathematical models from which novel results on brain and behavior have emerged. In this book the authors, who represent the cutting edge of this new discipline, draw connections between research in neuroeconomics and health economics. Moreover, they direct interested readers to a goldmine of additional references. Much of the research discussed here offers more questions than answers, and many of the answers are tentative first steps at a new understanding. It is precisely this that fuels our excitement about neuroeconomics. PMID- 19552303 TI - Emotion, decision-making and the brain. AB - PURPOSE: Initial explorations in the burgeoning field of neuroeconomics have highlighted evidence supporting a potential dissociation between a fast automatic system and a slow deliberative controlled system. Growing research in the role of emotion in decision-making has attempted to draw parallels to the automatic system. This chapter will discuss a theoretical framework for understanding the role of emotion in decision-making and evidence supporting the underlying neural substrates. DESIGN/METHODOLOGY/APPROACH: This chapter applies a conceptual framework to understanding the role of emotion in decision-making, and emphasizes a distinction between expected and immediate emotions. Expected emotions refer to anticipated emotional states associated with a given decision that are never actually experienced. Immediate emotions, however, are experienced at the time of decision, and either can occur in response to a particular decision or merely as a result of a transitory fluctuation. This chapter will review research from the neuroeconomics literature that supports a neural dissociation between these two classes of emotion and also discuss a few interpretive caveats. FINDINGS: Several lines of research including regret, uncertainty, social decision-making, and moral decision-making have yielded evidence consistent with our formulization- expected and immediate emotions may invoke dissociable neural systems. ORIGINALITY/VALUE: This chapter provides a more specific conceptualization of the mediating role of emotions in the decision-making process, which has important implications for understanding the interacting neural systems underlying the interface between emotion and cognition--a topic of immediate value to anyone investigating topics within the context of social-cognitive-affective neuroscience. PMID- 19552304 TI - Anxiety and decision-making: toward a neuroeconomics perspective. AB - PURPOSE: This chapter focuses on individual differences in anxiety, by reviewing its neurobiology, cognitive effects, with an emphasis on decision-making, and recent developments in neuroeconomics. METHODOLOGY: A review and discussion of anxiety and decision-making research. PRACTICAL IMPLICATIONS: This chapter argues that by making the step from emotional states to individual differences in emotion, neuroeconomics can extend its neurobiological roots and outreach its current clinical relevance. VALUE OF CHAPTER: This chapter contributes to the literature on individual differences in emotion and their effects on decision making, which is increasingly important in mainstream behavioral economics and neuroeconomics. PMID- 19552305 TI - The potential role of regret in the physician-patient relationship: insights from neuroeconomics. AB - PURPOSE: The aim of the chapter is to show how two important facts of physicians' behavior, (i) their tendency to "create" the demand for medical practices, and (ii) their delay and reluctance in using new treatments and therapies, can be explained with the lens of the neuroeconomics research on the neural and behavioral basis of regret. METHODOLOGY: This chapter adopts a neuroeconomics perspective on decision-making, asking how the brain represents values and generates emotional states, which consequently influence choices. In the line of recent work on emotion-based decision-making, we expect to be able to characterize the brain areas underlying the studied processes and to specify the functional relationship between rational decision-making and the emotional influences that modulate these decisional processes. ORIGINALITY: Neurobiological approaches can contribute significantly to a better understanding of the cognitive and emotional underpinnings of medical decision-making, from how physicians might evaluate and anticipate the effect of alternative therapies, to how patients might anticipate future consequences of their health choice. This can explain some features of the doctor-patient relationship which are not consistent with simple maximization models. FINDINGS: Our findings suggest that physicians' behavior can be often explained by regret avoidance. Likewise, they suggest that physicians play as actual agents when they make medical decisions that will affect the future well-being of their patients. RESEARCH LIMITATIONS: We limited our analysis to the potential role of anticipated regret; therefore, this chapter neglects many important factors of the health sector. PMID- 19552302 TI - Three studies on the neuroeconomics of decision-making when payoffs are real and negative. AB - PURPOSE: We summarize three previous neuroeconomic studies with two features that distinguish them from most others in experimental economics: (1) the use of physical pain to induce incentives and (2) acquisition of data on brain activation levels. By correlating behavior when payoffs are painful with brain activation, we are able to test for the neurobiological relevance of important phenomena previously observed in experimental studies that are at odds with classical economic theories of decision-making. These specific phenomena are (a) negative discounting of future payoffs; (b) nonlinear probability weighting; (c) the experience of regret and rejoice when making a decision under risk. METHODOLOGY/APPROACH: The expectation of pain is created through the use of mild electric shocks to the top of the foot. Pain confers disutility, so decisions are made in the domain of losses relative to the status quo. Simultaneous with these decisions, brain activation data is acquired through functional magnetic resonance imaging (fMRI). FINDINGS: We find evidence for negative time discounting of electric shocks. Participants who exhibited the most extreme forms of this discounting were distinguished by early and robust activation of a subset of the cortical pain matrix. We also find evidence for probability weighting in the domain of electric shocks, which is manifest at the neural level. We find evidence both behaviorally and neurally for regret and rejoice functions for painful outcomes. ORIGINALITY/VALUE OF CHAPTER: Previous experimental economic studies in the domain of losses have typically used monetary rewards. Here, we report behavioral effects and neural correlates using pain. PMID- 19552306 TI - How primates (including us!) respond to inequity. AB - PURPOSE: Responding negatively to inequity is not a uniquely human trait. Some of our closest evolutionary ancestors respond negatively when treated less well than a conspecific. Comparative work between humans and other primates can help elucidate the evolutionary underpinnings of humans' social preferences. METHODOLOGY/APPROACH: Results from studies of nonhuman primates, in particular chimpanzees and capuchin monkeys, are presented in comparison to human results that have been collected during economic game studies in humans, such as in the Ultimatum Game or Impunity Game. FINDINGS: Among nonhuman primates, a frequent behavioral reaction to inequity is to refuse to continue the interaction. While in some cases this response appears to be caused by the inequitable distribution, in others, it seems to be caused by another individual's inequitable behavior. While these reactions are similar to those of humans, this reaction does not appear to be a sense of fairness in the way that we think of it in humans. Neither nonhuman primate species alters their behavior when they are the benefited individual, and in an experimental situation, chimpanzees do not alter their behavior to obtain food for their partner as well as for themselves. ORIGINALITY/VALUE OF THE CHAPTER: Although there are differences between human and nonhuman primate responses, such studies allow us to better understand the evolution of our own responses to inequity. Given the strong behavioral reactions that even monkeys show to inequitable treatment, it is not surprising that humans are concerned with equity. Such comparisons increase understanding of issues such as healthcare disparities in humans. PMID- 19552307 TI - On the nature, modeling, and neural bases of social ties. AB - PURPOSE: This chapter addresses the nature, formalization, and neural bases of (affective) social ties and discusses the relevance of ties for health economics. A social tie is defined as an affective weight attached by an individual to the well-being of another individual ('utility interdependence'). Ties can be positive or negative, and symmetric or asymmetric between individuals. Characteristic of a social tie, as conceived of here, is that it develops over time under the influence of interaction, in contrast with a trait like altruism. Moreover, a tie is not related to strategic behavior such as reputation formation but seen as generated by affective responses. METHODOLOGY/APPROACH: A formalization is presented together with some supportive evidence from behavioral experiments. This is followed by a discussion of related psychological constructs and the presentation of suggestive existing neural findings. To help prepare the grounds for a model-based neural analysis some speculations on the neural networks involved are provided, together with suggestions for future research. FINDINGS: Social ties are not only found to be important from an economic viewpoint, it is also shown that they can be modeled and related to neural substrates. ORIGINALITY/VALUE OF THE CHAPTER: By providing an overview of the economic research on social ties and connecting it with the broader behavioral and neuroeconomics literature, the chapter may contribute to the development of a neuroeconomics of social ties. PMID- 19552308 TI - Emotion expression, decision-making and well-being. AB - PURPOSE: This chapter discusses the role of emotion expression in decision making. To understand connections between emotion and decision it is helpful first to differentiate between emotion experience and emotion expression. Understanding how emotion expression influences decision-making is important as a practical matter. However, in contrast to emotion experience, economic research has paid little attention to the significance of emotion expression in decision making. APPROACH: I review recent studies on emotion expression, paying specific attention to possible connections between emotion expression, punishment, fair economic exchange, and well-being. PRACTICAL IMPLICATIONS: In contrast to emotions, which are typically difficult to control, I suggest that opportunities for emotion expression can feasibly be manipulated through appropriately designed policies. I further suggest that this approach may have the ability to positively affect well-being and economic outcomes. VALUE OF THE CHAPTER: The chapter provides new perspectives on how policy-makers can benefit by understanding the effect of emotion expression in decision-making. The chapter also suggests future research to improve our understanding of emotion expression. PMID- 19552309 TI - Source preference and ambiguity aversion: models and evidence from behavioral and neuroimaging experiments. AB - PURPOSE: This experimental economics study using brain imaging techniques investigates the risk-ambiguity distinction in relation to the source preference hypothesis (Fox & Tversky, 1995) in which identically distributed risks arising from different sources of uncertainty may engender distinct preferences for the same decision maker, contrary to classical economic thinking. The use of brain imaging enables sharper testing of the implications of different models of decision-making including Chew and Sagi's (2008) axiomatization of source preference. METHODOLOGY/APPROACH: Using fMRI, brain activations were observed when subjects make 48 sequential binary choices among even-chance lotteries based on whether the trailing digits of a number of stock prices at market closing would be odd or even. Subsequently, subjects rate familiarity of the stock symbols. FINDINGS: When contrasting brain activation from more familiar sources with those from less familiar ones, regions appearing to be more active include the putamen, medial frontal cortex, and superior temporal gyrus. ROI analysis showed that the activation patterns in the familiar-unfamiliar and unfamiliar familiar contrasts are similar to those in the risk-ambiguity and ambiguity-risk contrasts reported by Hsu et al. (2005). This supports the conjecture that the risk-ambiguity distinction can be subsumed by the source preference hypothesis. RESEARCH LIMITATIONS/IMPLICATIONS: Our odd-even design has the advantage of inducing the same "unambiguous" probability of half for each subject in each binary comparison. Our finding supports the implications of the Chew-Sagi model and rejects models based on global probabilistic sophistication, including rank dependent models derived from non-additive probabilities, e.g., Choquet expected utility and cumulative prospect theory, as well as those based on multiple priors, e.g., alpha-maxmin. The finding in Hsu et al. (2005) that orbitofrontal cortex lesion patients display neither ambiguity aversion nor risk aversion offers further support to the Chew-Sagi model. Our finding also supports the Levy et al. (2007) contention of a single valuation system encompassing risk and ambiguity aversion. ORIGINALITY/VALUE OF CHAPTER: This is the first neuroimaging study of the source preference hypothesis using a design which can discriminate among decision models ranging from risk-based ones to those relying on multiple priors. PMID- 19552310 TI - Neuroeconomics of decision-making in the aging brain: the example of long-term care. AB - PURPOSE: Long-term care (LTC) services assist people with limitations in the ability to perform activities of daily living (ADLs) as a result of chronic illness or disabilities. We discuss possible behavioral explanations for the under-purchasing of LTC insurance, as well as the current state of knowledge on the neural mechanisms behind these behavioral factors. FINDINGS/APPROACH: Ideas from behavioral economics are discussed, including risk-seeking over losses, ambiguity-preferring over losses, hyperbolic discounting, and the effect of the aging process on the underlying neural mechanisms supporting these decisions. We further emphasize the role of age, as aging is a highly heterogeneous process. It is associated with changes in both brain tissue as well as cognitive abilities, and both are characterized by large individual differences. Therefore, understanding the neural mechanisms is vital to understanding this heterogeneity and identifying possible methods of interventions. RESEARCH IMPLICATIONS: LTC financing and insurance is a looming issue in the next 10-20 years. It is important to understand the process by which people make decisions about LTC insurance, heterogeneity in decision processes across individuals, and how these decisions interact with changes in policy and private LTC insurance markets. ORIGINALITY/VALUE OF THE CHAPTER: By providing an overview of the current state of knowledge in behavioral economics of LTC insurance and the neuroscience of aging, this chapter provides some new directions in the emerging area of neuroeconomics of aging. PMID- 19552311 TI - Health economic choices in old age: interdisciplinary perspectives on economic decisions and the aging mind. AB - PURPOSE: This chapter offers an integrative review of psychological and neurobiological differences between younger and older adults that might impact economic behavior. Focusing on key health economic challenges facing the elderly, it offers perspectives on how these psychological and neurobiological factors may influence decision-making over the life course and considers future interdisciplinary research directions. METHODOLOGY/APPROACH: We review relevant literature from three domains that are essential for developing a comprehensive science of decision-making and economic behavior in aging (psychology, neuroscience, and economics), consider implications for prescription drug coverage and long-term care (LTC) insurance, and highlight future research directions. FINDINGS: Older adults face many complex economic decisions that directly affect their health and well-being, including LTC insurance, prescription drug plans, and end of life care. Economic research suggests that many older Americans are not making cost-effective and economically rational decisions. While economic models provide insight into some of the financial incentives associated with these decisions, they typically do not consider the roles of cognition and affect in decision-making. Research has established that older age is associated with predictable declines in many cognitive functions and evidence is accumulating that distinct social motives and affect-processing profiles emerge in older age. It is unknown how these age differences impact the economic behaviors of older people and implies opportunities for path-breaking interdisciplinary research. ORIGINALITY/VALUE OF THE CHAPTER: Our chapter looks to develop interdisciplinary research to better understand the causes and consequences of age-related changes in economic decision-making and guide interventions to improve public programs and overall social welfare. PMID- 19552312 TI - Child health disparities, socio-economic status, and school enrollment decisions: evidence from German elementary school entrance exams. AB - PURPOSE: This chapter examines the role of child health for the intergenerational transmission of human capital. METHODOLOGY/APPROACH: The chapter uses unique administrative data from German elementary school entrance examinations. The chapter considers child health conditions such as obesity, low birth weight, ear problems, eye problems, behavioral problems, asthma, and allergies. We control for socio-economic and demographic characteristics of children and their parents as well as for institutional factors such as duration of pre-school attendance. FINDINGS: We find that health conditions are more common among children of less educated parents. We also find that health conditions have a substantially negative impact on school readiness, and the negative impact is considerably stronger for children of less-educated parents. In total, 55% of the school readiness gap can be attributed to health factors. Specifically, 19% of the gap can be attributed to differences in the prevalence of health conditions, and 36% of the gap can be attributed to differences in the severity of the impact. Thus, policies aimed at reducing disparities in child achievement should also focus on improving the health of disadvantaged children. ORIGINALITY: First, our study quantifies the extent to which the school readiness gap between parental education groups can be attributed to child health. Second, our data are of extraordinary quality, since they consist of a full sample of all children in one city and since they are collected during detailed examinations that were administered by government pediatricians. PMID- 19552314 TI - Expectations mediate objective physiological placebo effects. AB - PURPOSE: A placebo effect is a (positive) change in health outcomes that is due to a (positive) change in beliefs about the value of a treatment. Placebo effects might be "behavioral," in the sense that revised beliefs lead to behavioral changes or new actions that in turn yield changes in health outcomes. Placebo effects might also include a "physiological" component, which refers broadly to non-behavioral, brain-modulated mechanisms by which new beliefs cause changes in health outcomes. Nearly all formal economic models of human behavior are consistent with behavioral placebo effects, but strongly inconsistent with their physiological counterparts. The reason is that the latter effects can imply that expectations enter, rather than multiply, state-contingent preferences. It is therefore unfortunate that little evidence exists on physiological placebo effects. We report data from novel clinical experiments with caffeine that seek to provide such evidence. METHODS: Subjects visit the clinic on multiple occasions. On each visit they ingest either a placebo or caffeine pill. Subjects only know the probability with which the pill includes caffeine. We obtain physiological measurements prior to ingestion and at 30, 60, and 90 min after ingestion. Importantly, we constrain subjects to remain seated and read preselected magazines during the interval between treatment and outcome measurement. FINDINGS: Our design provides particularly clean inference because it (i) eliminates the possibility of behavioral confounds; (ii) provides for measurements at the individual level; (iii) manipulates beliefs without deception; and (iv) uses salient rewards. We find evidence for the existence of physiological placebo effects mediated by expectations. IMPLICATIONS: Our results are consistent with the possibility that the prefrontal cortex provides external, top-down control that modulates physiological outcomes, and make a case for the importance of research geared toward developing appropriate and tractable frameworks that accommodate non-linear relationships between expectations and preferences. PMID- 19552315 TI - The case for cases: a pedagogy for developing habits of thought. PMID- 19552313 TI - Temporal discounting as a measure of executive function: insights from the competing neuro-behavioral decision system hypothesis of addiction. AB - CONCEPTUAL PAPER PURPOSE: The purpose of this chapter is to examine a new conceptual model of addiction and interpret the results from delay discounting studies in light of this new perspective. METHODOLOGY/APPROACH: To accomplish this we (1) introduce this new conceptual model, (2) briefly review executive function, including evidence for executive dysfunction among the addicted, (3) describe the unique relationship of temporal discounting to the new model and executive dysfunction, and (4) reinterpret the discounting literature in light of this new conceptual model. FINDINGS: Addicted individuals discount the future more than controls. This is consistent with greater relative activation of the impulsive system and decreased relative activation of the executive system. It also supports the new conceptual model of addiction. RESEARCH IMPLICATIONS: The new model provides a model for understanding the observations from the broader area of research in temporal discounting. ORIGINALITY/VALUE OF CHAPTER: Given the view of executive function as important for the cross-temporal organization of behavior, we think that temporal discounting, the valuing of future commodities, qualifies this process to be included as an executive function. PMID- 19552316 TI - Evaluation tool for clinical nursing textbooks: bridging content analysis with evidence. AB - One of the challenges nurse educators face is choosing a textbook that ensures congruency within the discipline of nursing, national and global health priorities, and the mission of the university. This article discusses the development of a tool that evaluates course content concurrently with evidence. The need to critically link content analysis and evidence within clinical nursing textbooks is deemed important given the discipline's imperative to prepare nurses to use the best evidence available for practice. The history and concept of evidence-based nursing practice is explored to develop an operational definition for the tool, which was designed to guide a realistic and expeditious process for this important faculty responsibility. PMID- 19552317 TI - Examining patterns of change in the critical thinking skills of graduate nursing students. AB - Although critical thinking in undergraduate nursing education has been explored in depth, little is known about the critical thinking skills of graduate nursing students. Prior research on change in critical thinking scores is based primarily on pretest and posttest assessments that provide minimal information about change. This study used individual growth modeling to investigate how critical thinking skills change during a 2-year graduate nurse program. Scores from the evaluation, inference, and analysis subscales of the California Critical Thinking Skills Test comprised the empirical growth record. Change in the three critical thinking skills was more dynamic than that reported in previous studies. Patterns of change differed by critical thinking skill and in relation to students' initial critical thinking skill levels at program entry. PMID- 19552318 TI - Spiraling evidence-based practice and outcomes management concepts in an undergraduate curriculum: a systematic approach. AB - Preparing undergraduate nursing students to practice nursing in the 21st century requires a focus on the development of evidence-based practice and outcomes management knowledge and skills throughout the nursing curriculum. To this end, seven learning activities were created that spiral and increase in complexity while building on previously acquired skills. Working in teams and practicing team-building techniques, students learned how to develop a clinical question, search the literature, synthesize the current knowledge, identify the significance of the issue in an ecological model, decipher existing quality data and compare that data to national benchmarks, investigate a health care quality issue using quality improvement methods, and draft a proposal for implementation of a continuous quality improvement initiative. Work was presented in both written and oral presentation formats, with emphasis on engaging various audiences in a compelling health care issue. PMID- 19552319 TI - The nature of evidence in teaching practice. AB - A common thread in the educational literature is the recognition that teaching methods are appropriate for learners and that educators use the most effective teaching approaches. Unfortunately, the literature lacks a clear description of the evidence on which nurse educators base their teaching. Using a qualitative descriptive design, this study surveyed 14 nurse educators about the nature of the evidence they use in their teaching practices. Participants derived evidence from multiple sources and noted limited use of empirical research as a basis for teaching. An important consideration as to whether participants would modify their teaching was evaluation of that evidence. In addition, there were barriers and facilitators to the use of evidence. Based on the findings, a pluralistic view of evidence rather than any single source appears to be a preferable approach given the rapidly changing, dynamic nature of teaching practice and the nursing student population. PMID- 19552320 TI - Using new resources to teach evidence-based practice. AB - This article describes an innovative strategy used to teach evidence-based practice (EBP) concepts in an undergraduate research course and explains the underlying framework used to design the strategy. The strategy for the Evidence Based Practice Project was based on new resources: undergraduate nursing student competencies for EBP, national health care improvement priorities, evidence rating systems, and a model of knowledge transformation for EBP. Groups of students selected a priority area, categorized and critically appraised the evidence supporting the recommendation for change in health care practice, and compared the recommendation to actual practice. An oral and poster presentation of the project provided the opportunity to discuss the significance, influence, and strength of the evidence supporting the recommendation to change health care practice. Student presentations and faculty perceptions of the project indicated beginning competencies in EBP were achieved. PMID- 19552321 TI - Collaborative strategies for teaching evidence-based practice in accelerated second-degree programs. AB - An important challenge for teaching in accelerated second-degree programs is how to manage essential content within a compressed curriculum format. This article describes a project that used a collaborative model for teaching evidence-based practice (EBP) in a redesigned second-degree nursing program. Instructors in two courses shared responsibility for teaching basic concepts and guiding students' implementation of EBP in a clinical setting in partnership with clinical nurses. This approach resulted in a high degree of satisfaction for students, instructors, and nursing staff in clinical agencies. The project demonstrated collaborative teaching strategies can help students achieve basic knowledge in EBP and translate that knowledge into their clinical practice. Collaboration also can achieve more efficient learning experiences, a critical element in accelerated nursing programs. PMID- 19552322 TI - Student articulation of a nursing philosophical statement: an assignment to enhance critical thinking skills and promote learning. AB - The development of a personal philosophical statement about nursing is a useful assignment for both undergraduate and graduate nursing students. Students are asked to write a paper describing their philosophical views about nursing related to the metaparadigm concepts and include in-depth examples from their clinical practice to illustrate these views. This article includes information regarding important preparatory activities and handouts to facilitate the writing and evaluation of this assignment. There are many potential benefits of this complex assignment. It promotes students' reflection on their personal clinical practice experiences and engages students in mental and verbal dialogue that elicits their philosophical ideas about the metaparadigm concepts. These activities promote critical thinking and reflective clinical practice, act as a foundation for building theoretical understanding, and promote confidence in learning about nursing and other theories. PMID- 19552323 TI - Concept-based learning activities using the clinical judgment model as a foundation for clinical learning. AB - Several forces are influencing clinical education in the early 21st century. The clinical environment is changing as both patient acuity and patient-to-nurse staffing ratios increase. In many communities, the number of nursing students exceeds the number of clinical placements available. As nursing education moves forward, it is worthwhile to look more closely at how education occurs in the clinical environment regarding student progression from novice to more advanced levels of practice and how best to support development of student clinical judgment. Concept-based learning activities, first described by Heims and Boyd, are used to assist students in examining and addressing fundamental aspects (i.e., concepts) of pediatric patient and family care. The Clinical Judgment Model has been incorporated into concept-based learning activities as a framework to guide student thinking about the concepts. PMID- 19552324 TI - Addressing bugbears in practice while learning to read and synthesize research. PMID- 19552325 TI - Easing the pressure on A&E. PMID- 19552326 TI - Nurse practitioners. PMID- 19552327 TI - Developing a triage tool for paediatric care. AB - Most paediatric triage systems used in UK emergency departments (EDs) are adapted from adult tools. Paediatric-specific triage tools have been developed in Australia, Canada and the United States but there is no evidence of a similar tool in use in the UK despite support for its development. Staff at New Cross Hospital, in Wolverhampton, are involved in a project to develop a paediatric specific triage tool that can be used in EDs, assessment units and walk-in centres throughout the UK. The tool has been piloted in the hospital's paediatric assessment unit already with some success, but is yet to be validated. PMID- 19552328 TI - Transfer of war-zone casualties. AB - Many healthcare professionals in the UK are, like the author, also reservists in the armed forces and stationed in war zones overseas. In this article, the author describes the thrill of evacuating military and civilian personnel out of danger in Afghanistan. PMID- 19552329 TI - Launching a social enterprise see-and-treat service. AB - Many children who attend emergency departments with minor injuries or illnesses can be cared for by primary care services. This article describes an innovative partnership between a primary care trust and a social enterprise company to develop a see-and-treat primary care service that has reduced the number o children attending the traditional emergency department at a London hospital. PMID- 19552330 TI - Casting: complications and after care. AB - This article continues a short series of articles on how to apply casts. It examines some of the complications experienced by patients and discusses how emergency nurses can improve after-care services. PMID- 19552331 TI - Pistol crossbow injuries. AB - This article describes the wounding potential of pistol crossbows to prepare nurses who may have to provide immediate care for patients with injuries from these inexpensive and readily available weapons. PMID- 19552332 TI - After action reviews: a new model for learning. AB - University College London Hospitals NHS Foundation Trust is committed to developing a learning culture for its staff and, to achieve this, the organisation recently developed the after action review (AAR) model as a way for people involved in specific incidents to explore what happened and what they have learned. This article explains the concept of AAR and uses case studies to illustrate how it can improve patient care. PMID- 19552333 TI - Seizing the day. PMID- 19552334 TI - What is H1N1? PMID- 19552336 TI - How can case managers effectively handle stress-related illness? PMID- 19552337 TI - Workplace lactation programs: a critical element for breastfeeding mothers' success. AB - Employers play a critical role in mothers' success with breastfeeding when these women work full-time. Employers need information on the benefits breastfeeding can provide for working mothers and businesses. This article discusses current legislation and policies supporting mothers' ability to breastfeed in the workplace, the benefits of breastfeeding to the employer, mother, and child, and the need for employers to understand the benefits of breastfeeding. Finally, the role of occupational health nurses in facilitating the success of breastfeeding mothers through the development and implementation of a lactation program is explicated. PMID- 19552338 TI - Self-reported back pain among farm women in southeast Louisiana. AB - This cross-sectional survey details the incidence and predictors of self-reported back pain among farm women from southeastern Louisiana. The sample was obtained from a preexisting data set of a cross-sectional randomized sample of farm women who participated in the Louisiana Farm Family Injury Prevention and Health Initiative research project. The Haddon Injury Model was used as the conceptual framework. Approximately one fourth of the women in the sample reported back pain in the past year. The major findings from logistic regression analysis were that women reported back pain if they were hauling animals to market and had physical health conditions. Implications for nursing practice include assessing the risk of back pain among farm women and providing client education that includes the importance of compliance with recommended health screenings for early detection and treatment of physical health conditions associated with back pain. PMID- 19552340 TI - Image revisited. AB - The image that nurses project impacts their ability to influence others. Occupational health nurses who dress as other professionals at work dress will convince others that they can do more than treat injuries. PMID- 19552339 TI - The Family and Medical Leave Act: implications for occupational and environmental health nursing. AB - The Family and Medical Leave Act (FMLA) was enacted in 1993 to balance the demands of the workplace with the needs of families. Balancing work and family responsibilities will affect most workers as they experience their own serious illness or care for a child or a parent. The FMLA continues to present challenges regarding medical certifications, recordkeeping, intermittent leave management, and lack of understanding by employees and employers about rights and responsibilities under the law. This article discusses the rights and responsibilities of both parties. It also discusses how the occupational and environmental health nurse can bridge the gap between meeting the needs of the employee and those of the employer by serving as educator, advocate, and liaison/collaborator, leading to measurable cost savings for the employer and immeasurable benefits for the employee. PMID- 19552341 TI - Reduction in force perspectives--the occupational health nurse as the employee. AB - The occupational health nurse usually encounters reduction in force issues from a company perspective. This article discusses strategies for the occupational health nurse who is downsized. PMID- 19552342 TI - Hospital diversion scheme draws ire of national ED organizations. AB - The controversy over a new diversion policy at the University of Chicago Medical Center points out the pitfalls that can arise when seeking creative solutions to overcrowding and financial concerns, especially when it comes to compliance with the Emergency Medical Treatment and Labor Act (EMTALA). Here are some warnings offered by the experts: Your physicians must participate in all medical screening exams. You always can be second-guessed as to whether a case was "urgent." When conducting financial analyses, look at what visits actually cost you, not what you charge for care. PMID- 19552343 TI - New protocol slashes PCI-to-balloon time. AB - A new team-oriented process in Houston has enabled caregivers to dramatically reduce the time it takes for a patient to receive balloon angioplasty and, in the process, has vastly improved mortality rates. Here are some of their keys to success: As part of a "cocktail" given in the ambulance, the protocol calls for half-doses of retavase to reduce likelihood of reinfarction. Cardiologists are called from the ED once the process begins, so they often are in the ED or cath lab when the patient arrives. A set of criteria helps determine which patients will receive the new protocol. PMID- 19552344 TI - Improve your understanding of decision making's impact. PMID- 19552345 TI - Tips to succeed in getting sued. PMID- 19552346 TI - Standards on verbal orders rank high among common compliance problems. AB - The hectic pace of the ED makes it difficult to comply with The Joint Commission's standard on verbal orders, but some nurse managers have been able to increase compliance significantly with proactive strategies. Here are some of the successful approaches they have used: ongoing education and communication,including inservices, unit meetings, posters, and mass e-mails; incorporating the verbal orders standard into education about the National Patient Safety Goals; conducting random chart audits to gauge the level of staff compliance. PMID- 19552348 TI - We can fix health care with personalized HSAs. PMID- 19552347 TI - Uptake and inflammatory effects of nanoparticles in a human vascular endothelial cell line. AB - The mechanisms governing the correlation between exposure to nanoparticles and the increased incidence of cardiovascular disease remain unknown. Nanoparticles appear to cross the pulmonary epithelial barrier into the bloodstream, raising the possibility of direct contact with the vascular endothelium. Because endothelial inflammation is critical for the development of cardiovascular pathology, we hypothesized that direct exposure of human aortic endothelial cells (HAECs*) to nanoparticles induces an inflammatory response and that this response depends on the composition of the particles. To test this hypothesis, we incubated HAECs for 1 to 8 hours with different concentrations (0.001-50 microg/mL) of iron oxide (Fe2O3), yttrium oxide (Y2O3), cerium oxide (CeO2), and zinc oxide (ZnO) nanoparticles. Using real-time reverse transcriptase-polymerase chain reaction (RT-PCR), we subsequently measured messenger RNA (mRNA) levels of three markers of inflammation: intercellular cell adhesion molecule-1 (ICAM-1), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1). The particles were well characterized in terms of size, surface area, composition, and crystal structure. To determine the interactions of nanoparticles with HAECs, we used inductively coupled plasma-mass spectrometry (ICP-MS) to measure the concentration of internalized particles. Our data indicate that the delivery of nanoparticles to the HAEC surface and their uptake within the cells correlate directly with the concentration of particles in the cell culture medium. Transmission electron microscopy (TEM) revealed that the Fe2O3, Y2O3, and ZnO nanoparticles are internalized by HAECs and are often found within intracellular vesicles (the CeO2 particles were not imaged). Fe2O3 nanoparticles did not provoke an inflammatory response in HAECs at any of the concentrations tested, CeO2 particles elicited no response at low concentrations and a weak response above 10 microg/mL, and Y2O3 and ZnO nanoparticles elicited a pronounced inflammatory response above a threshold concentration of 10 microg/mL. We used fluorescent markers to identify the production of reactive oxygen species (ROS) in cells; the results showed that Y2O3 and ZnO particles at the highest concentrations may lead to the production of ROS. At the highest concentration, ZnO nanoparticles caused significant loss of cell adherence. These results demonstrate that inflammation in HAECs after acute exposure to metal oxide nanoparticles depends on the concentration and composition of the particles. PMID- 19552349 TI - The physician's role: overcoming challenges to serve in the best interest of the patient. PMID- 19552350 TI - Meeting the research infrastructure needs of micropolitan and rural communities. AB - In the 1800s, this country chose to establish land-grant colleges to see that the working class could attain higher education, and that the research needs of the agricultural and manufacturing segments of this country could be met. It seems contrary to our origins to see so little support at present for research infrastructure going to the very communities that need such research to sustain their populations, grow their economies, to attract physicians, to provide adequate health care, and to educate, retain, and employ their youth. Cities are viewed as sources for high-paying jobs, yet many of these same jobs could be translated to rural and micropolitan areas, provided that the resources are established to support it. One of the fastest growing economic periods in this country's history was during World War II, when even the smallest and most remote towns contributed substantially to the innovations, manufacture, and production of goods benefiting our nation as a whole. Rural areas have always lagged somewhat behind metropolitan areas in acquisition of new technology. Rural electricity and rural phone access are examples from the past. Testing our universities' abilities to grow distributive research networks beyond their campuses will create a competitive edge regionally, against global workplace, educational, and research competition, and will lay the groundwork for efficiency in research and for new innovation. PMID- 19552351 TI - Physician perspectives on quality and error in the outpatient setting. AB - CONTEXT: Little is known about the influence of the primary care workplace on patient care. Assessing physician opinion through focus groups can elucidate factors related to safety and error in this setting. METHOD: During phase 1 of the Minimizing Error, Maximizing Outcome (MEMO) Study, 9 focus groups were conducted with 32 family physicians and general internists from 5 areas in the upper Midwest and New York City. RESULTS: The physicians described challenging settings with rapidly changing conditions. Patients are medically and psychosocially complex and often underinsured. Communication is complicated by multiple languages, time pressure, and inadequate information systems. Complex processes of care have missing elements including medication lists and test results. Physicians are pressed to be more productive, and key administrative decisions are made without their input. Targeted areas to improve safety and reduce error included teamwork, aligned leadership values, diversity, collegiality, and respect. CONCLUSIONS: Primary care physicians clearly described positive and negative workplace factors related to safety and error. The themes suggest that systems of care and their dynamic nature warrant attention. Enhancing positive and ameliorating negative cultures and processes of care could bring real benefits to patients, physicians, and ambulatory office settings. PMID- 19552353 TI - Factors associated with the seasonality of blood lead levels among preschool Wisconsin children. AB - CONTEXT: Aggregate blood lead testing data for Wisconsin children younger than age 6 exhibit seasonal trends in both average blood lead levels and in the percent of those tested and found to have blood lead levels in excess of the 10 mcg/dL threshold for poisoning. Blood lead levels and poisoning rates typically peak during the late summer and early fall months, and are at their minimum during the late winter. METHOD: Blood test data was analyzed to determine variations by month and age. RESULTS: Seasonal variations are evident even among the very young: infants younger than 10 months who likely have limited opportunity to encounter lead hazards within their home or in the outdoor soil. Seasonal periodicity is most evident among infants who reside in very urban and very rural communities. The observed seasonal periodicity might be associated with the seasonal availability of lead within the children's environment. Particulate matter data measured at several ambient air quality monitoring stations exhibit a similar periodic seasonality, suggesting a possible relationship between blood lead levels and the availability of dust and airborne particulates during the summer months. CONCLUSION: Clinicians should consider the seasonality of blood lead levels when scheduling tests and interpreting test results. PMID- 19552354 TI - Emergency contraception and physicians' rights of conscience: a review of current legal standards in Wisconsin. AB - Recent legislation in Wisconsin mandating provision of emergency contraception to victims of sexual assault may create a conflict of conscience for some health care professionals. Although disputes exist over the exact mechanism of action of emergency contraception, those professionals who espouse a particularly strict stance may be reluctant to dispense the medication for fear that it could prevent a fertilized embryo from implanting in the uterus. While no objection of conscience clause was written into the new law, Wisconsin law has a long tradition of recognizing rights of conscience in matters of religious conflict. This legal tradition both at statutory and common law levels is summarized with application to the recent emergency contraception mandate. A case is made for a potential legal defense should a health care professional abstain from dispensing emergency contraception. PMID- 19552355 TI - Transforming medicine from the bottom up. PMID- 19552352 TI - The prescribing patterns of Wisconsin family physicians surrounding saline nasal irrigation for upper respiratory conditions. AB - CONTEXT: Upper respiratory conditions are common and have a significant impact on patient quality of life, medical resource expenditure, and antibiotic use. Saline nasal irrigation (SNI) is an adjunctive therapy for upper respiratory conditions; clinical studies suggest that use of SNI may be effective for symptoms of upper respiratory conditions, and its popularity seems to be growing. The prescribing patterns of physicians regarding SNI have not yet been studied. OBJECTIVE: To assess the use among family physicians in Wisconsin of SNI, determine how and for which conditions they recommend SNI, and the degree to which they experience clinical success with SNI. METHOD: This was assessed by an electronic questionnaire of 330 practicing family physicians in the Wisconsin Academy of Family Physicians and Wisconsin Research and Education Network. RESULTS: Analysis showed 286 of 330 respondents (87%) have used SNI as adjunctive care for a variety of upper respiratory conditions including chronic rhinosinusitis (91%), acute bacterial rhinosinusitis (67%), seasonal allergic rhinitis (66%), viral upper respiratory infection (59%), other allergic rhinitis (48%), irritant-based congestion (48%), and rhinitis of pregnancy (17%). Respondents also reported having used SNI prior to antibiotics for acute bacterial rhinosinusitis (77%). Use patterns varied regarding type of SNI administration, dosing frequency, saline concentration, and patient education. CONCLUSIONS: This questionnaire based study suggests that SNI is used by family physicians for a variety of upper respiratory conditions though recommendation and patient education styles, dosing schedules, and solution types vary. PMID- 19552356 TI - Having a clear vision. PMID- 19552357 TI - Calcium channel blockers and gingival hyperplasia--an update. PMID- 19552358 TI - The use of direct gold in restorative dentistry. AB - Direct gold is not the most utilized dental restorative technique; however, as Frederick Eichmiller notes, no other material allows the marginal gap to approach 0 micro (personal communication, 2005). Placing gold foils is best learned in an operating study club environment with an experienced mentor. The technique is rarely (if ever) taught in any dental school, although a few schools have electives that expose students to this classic method of operative dentistry. PMID- 19552359 TI - Attachment-retained overdentures: the most cost-effective implant solution for the edentulous arch. PMID- 19552360 TI - Curing lights--does a 5-second cure really work? PMID- 19552361 TI - Management of dentinal hypersensitivity. AB - Dentinal hypersensitivity is a complex condition that can cause considerable concern in the dental office. Despite the large number of available treatment modalities, no current desensitizing agent is considered ideal for managing this uncomfortable condition. Selecting the right therapy requires a complete understanding of how applying a stimulus to the exposed dentin surface can influence the nerve fibers and produce hyperesthesia. This article reviews the etiology and critically analyzes management of dentinal hypersensitivity by reviewing laboratory and clinical investigations. PMID- 19552362 TI - Residual (ghost) sockets in bisphosphonate use--evidence of poor healing and slow bone turnover. AB - Patients taking bisphosphonate drug therapy have demonstrated extremely poor alveolar bone healing after relatively minor oral surgical procedures. It would seem logical that extraction sockets could remain visible radiographically for an extended period after surgery, even in cases with soft tissue healing. This article chronicles the case of a patient who had been taking zoledronic acid chronically for metastatic cancer and who demonstrated numerous residual sockets (also known as ghost sockets), with lamina dura outlines that were visible radiographically. PMID- 19552363 TI - Factors involved in microbial colonization of oral prostheses. AB - The presence of biofilm or denture plaque on the tissue contacting the (intaglio) surface of a denture is a major etiologic factor in the pathogenesis of both denture stomatitis and inflammatory papillary hyperplasia. This article reviews the literature concerning the various factors that contribute to the development of denture plaque and its colonization by Candida albicans and other microorganisms. PMID- 19552364 TI - Influence of gel color on in vitro surface and intrapulpal temperature rise during blue light-activated tooth bleaching. AB - This study evaluated the null hypothesis that the color of a bleaching gel has no effect on temperature because it absorbs the light used during photocuring. This study used 10 maxillary central incisors (offering endodontic access) and a digital thermometer with thermocouples. One end of the thermocouple was placed on the labial surface (immersed in bleaching gel); the other end was placed in the pulp chamber. The bleaching materials each offered two color options: blue and red. The bleaching agents were activated with a LED hybrid blue light photocuring unit and a low intensity diode laser. Based on the blue light-activated bleaching, it was determined that the red gel produced significantly higher temperature increases than the blue gel. No significant differences were found between the two brands of gel tested. PMID- 19552365 TI - Dental erosion and acid reflux disease: an overview. AB - Dental erosion can be difficult to detect, especially in the early stages when lesions are subtle and can be easily overlooked. Patients often are not aware of erosion until the dentition has sustained severe damage that requires extensive and expensive dental rehabilitation. The pH of stomach acid is much lower than the critical pH of enamel dissolution; therefore, reflux of stomach contents into the oral cavity over an extended period of time can cause severe loss of tooth structure. Dental treatment for reflux-induced erosion should focus not only on appropriate restoration but also on all available preventive measures, such as neutralization of acid and remineralization or strengthening of enamel against acid attack. Dentists must maintain a high degree of suspicion for reflux-induced erosion whenever a patient displays symptoms of acid reflux disease or a pattern of erosion that suggests an intrinsic source of acid exposure. PMID- 19552366 TI - Opportunities for practicing non-invasive dentistry during the process of routine crown preparation. AB - Minimally invasive dentistry involves the philosophy of sacrificing minimal normal tissue to preserve a maximal natural tooth volume. Preparing a tooth for a crown exposes the adjacent proximal surfaces; these can be inspected carefully to determine a need for minimally invasive repair. Small, cavitated lesions can be identified and resolved in an early stage of disease. PMID- 19552367 TI - Influence of matrix and insertion technique on the microleakage and microhardness of posterior composite restorations. AB - This study sought to evaluate how the matrix and insertion technique affected microleakage and microhardness of posterior composite restorations. Class II preparations with the gingival margin in cementum were performed in 60 bovine teeth and divided into two groups: transparent matrix and reflective wedge (Group T) (n=20) and metallic matrix and wood wedge (Group M) (n=40). Cavities were restored and specimens were isolated with nail varnish, immersed in 0.2% Rhodamine B, and sectioned. Microleakage was evaluated under magnification (20x) and data were analyzed with one-way ANOVA. Microhardness was assessed at 1, 2, and 3 mm from the gingival wall and data were analyzed with two-way ANOVA. No significant difference was observed in terms of leakage. Significant differences were observed in terms of hardness for both factors, although no interaction was noted. It was concluded that the insertion technique had no influence on microleakage and that using a metallic matrix with oblique increments provided the highest hardness. There was no correlation between these two variables. PMID- 19552368 TI - Comparison of the fit of Procera crowns made from stone with those made from polyurethane resin. AB - This study compared the average marginal gap of Procera crowns fabricated from a definitive cast made from polyurethane resin with those made from type IV stone. A metallic maxillary central incisor and first premolar were prepared according to the Procera guidelines and incorporated into a U-shaped definitive cast. From this definitive cast, 20 primary casts were made in polyurethane resin. Two sets of impressions (10 with type IV stone and 10 with polyurethane resin) were made and poured onto these casts. Forty pure aluminum oxide substructures for Procera crowns were made for these groups, with 10 incisors and 10 premolars in each group. Each substructure was cemented on the respective primary die and sectioned. Using an optical microscope and special software, sections were measured and compared for marginal adaptation. Based on the findings, both materials were equally adequate for producing a Procera crown. However, the mean margin gap was consistently larger for type IV stone dies than for polyurethane resin dies. PMID- 19552369 TI - Parotid swelling and facial nerve palsy: an uncommon presentation of sarcoidosis. AB - Sarcoidosis is a granulomatous disease of unknown etiology that can affect virtually any organ system. Diagnosing sarcoidosis can be challenging, since manifestations of the disease will vary depending on the organ systems involved. The present case describes a 24-year-old woman with bilateral jaw pain and swelling, who developed bilateral facial nerve palsies secondary to sarcoidosis. Consideration of sarcoidosis in the differential diagnosis of facial nerve palsy or parotid swelling is important to ensure proper treatment. PMID- 19552370 TI - Unusual gingival pigmentation. Intentional tattooing. PMID- 19552371 TI - Crusted facial lesion of two years' duration. Basosquamous carcinoma. PMID- 19552372 TI - E-novo: an automated workflow for efficient structure-based lead optimization. AB - An automated E-Novo protocol designed as a structure-based lead optimization tool was prepared through Pipeline Pilot with existing CHARMm components in Discovery Studio. A scaffold core having 3D binding coordinates of interest is generated from a ligand-bound protein structural model. Ligands of interest are generated from the scaffold using an R-group fragmentation/enumeration tool within E-Novo, with their cores aligned. The ligand side chains are conformationally sampled and are subjected to core-constrained protein docking, using a modified CHARMm-based CDOCKER method to generate top poses along with CDOCKER energies. In the final stage of E-Novo, a physics-based binding energy scoring function ranks the top ligand CDOCKER poses using a more accurate Molecular Mechanics-Generalized Born with Surface Area method. Correlation of the calculated ligand binding energies with experimental binding affinities were used to validate protocol performance. Inhibitors of Src tyrosine kinase, CDK2 kinase, beta-secretase, factor Xa, HIV protease, and thrombin were used to test the protocol using published ligand crystal structure data within reasonably defined binding sites. In-house Respiratory Syncytial Virus inhibitor data were used as a more challenging test set using a hand-built binding model. Least squares fits for all data sets suggested reasonable validation of the protocol within the context of observed ligand binding poses. The E-Novo protocol provides a convenient all-in-one structure-based design process for rapid assessment and scoring of lead optimization libraries. PMID- 19552373 TI - Synthesis of magnetic Fe2O3/Au core/shell nanoparticles for bioseparation and immunoassay based on surface-enhanced Raman spectroscopy. AB - Magnetic Fe2O3/Au core/shell nanoparticles can be particularly used in biological separation, but the development of an appropriate technique including a production process for higher efficient separation and the subsequent immunoassay for lower level still represent a great challenge. In this article, Fe2O3/Au core/shell nanoparticles with different Au ratios were prepared by reducing HAuCl4 on the surface of gamma-Fe2O3 nanoparticles. Scanning electron microscopy (SEM) images and surface-enhanced Raman spectroscopy (SERS) spectra clearly show that the surfaces of Fe2O3 nanoparticles were covered by Au. SERS signals of pyridine (Py) have been obtained on the Fe2O3/Au nanoparticles, and it has been found that the SERS intensity enhanced with the increase of iterative additions of HAuCl4. The antigens in test solution have been effectively separated by the magnetic Fe2O3/Au core/shell nanoparticles, and subsequent rapid detection was examined by immunoassay analysis based on SERS. The result demonstrates that the magnetic bioseparation program used by this magnetic Fe2O3/Au core/shell nanoparticles could separate almost all of the antigens in test solution. The ease of operation and good separation efficiency of this effective method has shown a potential application for magnetic Fe2O3/Au core/shell nanoparticles in bioseparation. PMID- 19552375 TI - Chloroalkane gel formations by tris-urea low molecular weight gelator under various conditions. AB - C3-symmetrical tris-urea low molecular weight gelator 1 was synthesized in three steps from phloroglucin. The gelation ability of 1 in four chloroalkanes, i.e., CH2Cl2, CHCl3, 1,1,2-trichloroethane, and 1,1,2,2-tetrachloroethane, was investigated under various conditions. Thermal treatment was effective in gelating 1,1,2-trichloroethane. In the presence of equimolar 1 and CuBr2, 1,1,2 trichloroethane and 1,1,2,2-tetrachloroethane formed gels. Mixtures of 1 and CHCl3 or 1,1,2,2-tetrachloroethane gave gels after ultrasound irradiation. CH2Cl2 changed into a gel in the presence of equimolar 1 and BiCl3 after sonication. Spherical particles with rough surfaces were found by SEM observation of CHCl3 gel prepared from ultrasound irradiation of 1 and Y(NO3)3. PMID- 19552374 TI - Polymer-mesoporous silica materials templated with an oppositely charged surfactant/polymer system for drug delivery. AB - In this paper, polymer-mesoporous silica nanoparticles have been synthesized by a dual-template technology. Cationic polymer quaternized poly[bis(2 chloroethyl)ether-alt-1,3-bis[3-(dimethylamino)propyl]urea] (PEPU) and anionic surfactant sodium dodecyl sulfate (SDS) were used to form a homogeneous comicelle system to induce mesoporous silica spherical nanoparticles with diameters of 50 180 nm. The formation mechanism was studied by transmission electron microscopy (TEM), which suggested that PEPU played a cotemplate role in the synthesis process, and no mesoporous structure was generated without it. After removing the anionic surfactant, SDS, by an ion-exchange method, the cationic polymer mesoporous silica nanoparticles were obtained. Using the materials as the host and ibuprofen (IBU)/captopril (CapH(2)) as the model drugs, the system revealed well-sustained release profiles. PMID- 19552378 TI - Pd(PPh3)4-PEG 400 catalyzed protocol for the atom-efficient Stille cross-coupling reaction of organotin with aryl bromides. AB - Aryl bromides (4 equiv) were coupled efficiently with organotin (1 equiv) in an atom-efficient way using the tetra(triphenylphosphine)palladium/polyethylene glycol 400 (Pd(PPh3)4/PEG 400) catalytic system in the presence of sodium acetate (NaOAc) as base at 100 degrees C, providing excellent yields of the corresponding functionalized biaryls in short reaction times. PMID- 19552377 TI - Discovery and development of a small molecule library with lumazine synthase inhibitory activity. AB - (E)-5-Nitro-6-(2-hydroxystyryl)pyrimidine-2,4(1H,3H)-dione (9) was identified as a novel inhibitor of Schizosaccharomyces pombe lumazine synthase by high throughput screening of a 100000 compound library. The K(i) of 9 vs Mycobacterium tuberculosis lumazine synthase was 95 microM. Compound 9 is a structural analogue of the lumazine synthase substrate 5-amino-6-(d-ribitylamino)-2,4 (1H,3H)pyrimidinedione (1). This indicates that the ribitylamino side chain of the substrate is not essential for binding to the enzyme. Optimization of the enzyme inhibitory activity through systematic structure modification of the lead compound 9 led to (E)-5-nitro-6-(4-nitrostyryl)pyrimidine-2,4(1H,3H)-dione (26), which has a K(i) of 3.7 microM vs M. tuberculosis lumazine synthase. PMID- 19552379 TI - Enantio- and diastereoselective asymmetric addition of 1,3-dicarbonyl compounds to nitroalkenes in a doubly stereocontrolled manner catalyzed by bifunctional rosin-derived amine thiourea catalysts. AB - Starting from commercially available natural rosin derivatives, a class of bifunctional rosin-derived amine thiourea catalysts were designed and synthesized. The doubly stereocontrolled asymmetric addition of a variety of 1,3 dicarbonyl compounds to nitroalkenes was investigated. These rosin-derived chiral thioureas have been shown to serve as effective catalysts for this double sterecontrolled organocatalytic process by the investigation of the efficacy of the thiourea catalysts in comparison with other thiourea catalysts reported. In addition, these chiral thiourea ligands are easily available. Furthermore, the rosin-derived tertiary amine-thiourea was also revealed to be highly efficient for construction of contiguous stereogenetic centers containing an asymmetric quaternary carbon by the Michael reaction of alpha-substituted beta-ketoesters to nitroalkenes. PMID- 19552380 TI - Concise and versatile multicomponent synthesis of multisubstituted polyfunctional dihydropyrroles. AB - Tetra- and pentasubstituted polyfunctional dihydropyrroles have been concisely synthesized in high yields by two different processes of the one-pot multicomponent reactions (MCRs) of but-2-ynedioates 1, amines 2, and aldehydes 3 at room temperature or at 70 degrees C. The first one involves a domino hydroamination/nucleophilic addition/amidation-cyclization process and leads to the formation of tetrasubstituted polyfunctional dihydropyrroles 4. The second undergoes hydroamination/amidation/intramolecular cyclization/imine-enamine tautomerization sequence and results in pentasubstituted products 5. The structures of 4 and 5 were confirmed by single-crystal X-ray diffraction. These novel methodologies provide easy access to diversely multisubstituted polyfunctional dihydropyrrole libraries. The primary biological screening in vitro against HIV-1 has shown that 22 tested compounds have exhibited significant activity with IC(50) in micromolar range (38-58 microM). PMID- 19552381 TI - Single-crystal-to-single-crystal transformation in a one-dimensional Ag-Eu helical system. AB - Single-crystal-to-single-crystal transformation of one-dimensional 4d-4f coordination polymers has been investigated for the first time. More importantly, we observed the transformation of a meso-helical chain to a rac-helical chain as a function of the temperature. PMID- 19552382 TI - A perspective distilled from seventy years of research. AB - Physical organic chemistry might be regarded as officially recognized as a distinct discipline through the publication of L. P. Hammett's book of that title, although substantial earlier work can be traced back to the turn of the 20th century. Many of the instrumental tools that helped the discipline develop in so many different ways began to appear in the late thirties and during World War II and were soon built to be increasingly operated in the "hands-on" mode. This development became very popular in academia, where instruments are not operated for you by an expert, but even if you are an undergraduate, you can more or less be the expert yourself and take many varieties of data on instruments usually available on a 24 h basis. It has been my privilege and joy to begin research in chemistry just as these waves of change began to grow and to savor the great contribution that the new methods, such as measurement of (14)C, UV vis, IR, NMR, and hands-on use of computers, made in facilitating our research programs at MIT and later at Caltech. Among those programs, which will be discussed, were (14)C tracing of carbocation rearrangements and benzyne formation, electrical effects of substituents, Grignard reagents, synthesis of small-ring compounds, (2 + 2) cycloaddition reactions of halogenated ethylenes, assisting in development of (19)F, (13)C, and (15)N NMR for conformational analysis, other structural, kinetic, and tracer studies, as well as helping through textbooks to bring Huckel MO theory and the elements of NMR to familiarity for organic chemists. From the very beginning of my research career, I have been the beneficiary of personal mentoring which has been very crucial to my success in research and is an important theme in what follows. PMID- 19552383 TI - Silver Nanowire Networks as Flexible, Transparent, Conducting Films: Extremely High DC to Optical Conductivity Ratios. AB - We have used aqueous dispersions of silver nanowires to prepare thin, flexible, transparent, conducting films. The nanowires are of length and diameter close to 6.5 MUm and 85 nm, respectively. At low thickness, the films consist of networks but appear to become bulk-like for mean film thicknesses above ~160 nm. These films can be very transparent with optical transmittance reaching as high as 92% for low thickness. The transmittance (550 nm) decreases with increasing thickness, consistent with an optical conductivity of 6472 S/m. The films are also very uniform; the transmittance varies spatially by typically <2%. The sheet resistance decreases with increasing thickness, falling below 1 Omega/? for thicknesses above 300 nm. The DC conductivity increases from 2 * 10(5) S/m for very thin films before saturating at 5 * 10(6) S/m for thicker films. Similarly, the ratio of DC to optical conductivity increases with increasing thickness from 25 for the thinnest films, saturating at ~500 for thicknesses above ~160 nm. We believe this is the highest conductivity ratio ever observed for nanostructured films and is matched only by doped metal oxide films. These nanowire films are electromechanically very robust, with all but the thinnest films showing no change in sheet resistance when flexed over >1000 cycles. Such results make these films ideal as replacements for indium tin oxide as transparent electrodes. We have prepared films with optical transmittance and sheet resistance of 85% and 13 Omega/?, respectively. This is very close to that displayed by commercially available indium tin oxide. PMID- 19552385 TI - D-ring-opened phragmalin-type limonoid orthoesters from the twigs of Swietenia macrophylla. AB - Sixteen new D-ring-opened phragmalin limonoid orthoesters, swietenitins A-M (1-6, 8, 10, 12-16), 2-acetoxyswietenialide D (7), 2,11-diacetoxyswietenialide D (9), and 11-deoxyswietenialide D (11), and four known compounds were isolated from the twigs of Swietenia macrophylla. The structures of 1-16 were established on the basis of spectroscopic methods, and 1 and 2 were confirmed by single-crystal X ray diffraction. The (1)H NMR-based conformational analysis on the epimeric compounds 1 and 2 provided a general approach to determine the absolute configuration of the 2,3-epoxy-2-methylbutyryloxy unit at C-3 borne by a large group of the known phragmalin-type limonoid orthoesters. PMID- 19552384 TI - Combined chemical and biosynthetic route to access a new apoptolidin congener. AB - Glycosylation of a synthetic aglycone using precursor-directed biosynthesis is facilitated by a chemical ketosynthase "knockdown" of the apoptolidin producer Nocardiopsis sp. This synthetic approach facilitated the preparation of an unnatural disaccharide derivative of apoptolidin D that substantially restores cytotoxicity against H292 cells and deconvolutes the role of the decorating sugars in apoptolidin bioactivity. PMID- 19552386 TI - Fabrication, self-assembly, and properties of ultrathin AlN/GaN porous crystalline nanomembranes: tubes, spirals, and curved sheets. AB - Ultrathin AlN/GaN crystalline porous freestanding nanomembranes are fabricated on Si(111) by selective silicon etching, and self-assembled into various geometries such as tubes, spirals, and curved sheets. Nanopores with sizes from several to tens of nanometers are produced in nanomembranes of 20-35 nm nominal thickness, caused by the island growth of AlN on Si(111). No crystal-orientation dependence is observed while releasing the AlN/GaN nanomembranes from the Si substrate indicating that the driving stress mainly originates from the zipping effect among islands during growth. Competition between different relaxation mechanisms is experimentally revealed for different nanomembrane geometries and well described by numerical calculations. The cathodoluminescence emission from GaN nanomembranes reveals a weak peak close to the GaN bandgap, which is dramatically enhanced by electron irradiation. PMID- 19552388 TI - Stereoselective quaternary center construction via atom-transfer radical cyclization using silicon tethers on acyclic precursors. AB - A novel strategy for the stereoselective construction of all-carbon quaternary centers on acyclic molecules using a two-step tandem process is reported. The first step involves an intramolecular and stereoselective atom transfer radical cyclization reaction from an allyl or vinyl subunit attached on a silyloxy, serving as a tether, to a tertiary radical alpha to an ester. A subsequent mild acidic elimination leads stereoselectively to a quaternary center bearing an allyl or a vinyl in high yield. PMID- 19552387 TI - Stereoselective cross-coupling between allylic alcohols and aldimines. AB - A cross-coupling reaction between an allylic alcohol and an imine is described for stereoselective allylation of aromatic and aliphatic imines. This method provides operationally simple, enantioselective access to functionalized homoallylic amines. Particularly noteworthy is direct use of a functionalized allylic alcohol as an allylating reagent without prederivatization, which obviates the use of preformed organometallic reagents or activated imine derivatives. PMID- 19552389 TI - Polynuclear nickel(II) complexes: a magnetostructural study of Ni(II)4, Ni(II)6, and Ni(II)9 species with oxime ligands. AB - Syntheses, crystal structures, and magnetic properties are reported for a tetrametallic (complex 1), a nonametallic (complex 2), and two hexametallic (complexes 3 and 4) nickel(II) clusters, namely, [Ni(4)(HL(1))(2)(mu OAc)(2)(MeOH)] (1), [Ni(9)(L(2))(10)(mu(3)-OH) (2) (mu-OH) (2) (mu-OH (2))(2)(OH(2))(6)](ClO(4)) (4) (2), [Ni(6)(L(2))(9)(L(2)H)(MeOH)(H(2)O)(2)](ClO(4))(3) (3), and [Ni(6)(L(3))(3)(mu(3) O)(2)](ClO(4))(2) (4), where H(4)L(1) represents N,N'-dimethyl-N,N'-ethylene bis(5-bromo-3-formyloxime-2-hydroxybenzylamine); HL(2), 1-methylimidazole-2 aldoxime; and H(2)L(3), N,N'-bis(2,3-butanedionemonoxime-2-ene) 3(aminomethyl)benzylamine. The structure of 1 can be considered as two face sharing bioctahedral units of [Ni(2)(mu-O(phen))(2)(mu-OAc)] bridged by a two atom (-N-O-) oximate linker. The Ni(II)...Ni(II) distances of av. 2.935 A preclude metal-metal bonding, although they are remarkably short. Variable temperature magnetic susceptibility data are fitted to obtain the parameters J (1) = +8.0 cm(-1), J(2) = -16.0 cm (-1), and g = 2.19 (H= -2 JS(i) x S(j)). The ferromagnetic coupling J (1) operates between the nickel(II) centers in the face sharing bioctahedral units, whereas J(2) represents the antiferromagnetic interactions mediated by a single (-N-O-) bridge separating the two nickel centers at a distance of approximately 4.71 A. A rationale for the disparate nature of interactions based on a comparison with those reported in the literature is forwarded. The structure of 2 consists of two [Ni(4)(L(2))(5)] units linked covalently to a central nickel atom by four oximate and two hydroxy oxygen atoms, resulting in a central octahedral NiO(6) core and thus yielding the nonanuclear nickel(II) cluster. The magnetic data were analyzed by a "two-J" model, yielding pairwise antiferromagnetic exchange interactions, J(1) = -24.0 cm (-1) and J(2) = -5.8 cm (-1), between the nickel centers. The spin ground state of S(t) = 1.0 has been confirmed by magnetization measurements (variable temperature, variable-field) at different fields. The structure of 3 contains six nickel(II) centers, each of which is six-coordinated but with different coordination environments: NiN(6), NiO(6), NiN(3)O(3)(2x), NiN(4)O(2), and NiNO(5). The ground-state spin has been observed to be S(t) = 1.0 with the axial zero-field splitting parameter D = -7.2 cm(-1). Complex 4 is a rare example of dimeric [Ni(3)(mu(3)-O)](4+) units, in which each of six nickel(II) centers is in square-planar geometry with low-spin d(8) Ni(II) centers, thus rendering diamagnetism to complex 4. PMID- 19552390 TI - Cyclization reactions through DDQ-mediated vinyl oxazolidinone oxidation. AB - Vinyl oxazolidinones react with DDQ to form alpha,beta-unsaturated acyliminium ions in a new method for forming electrophiles under oxidative conditions. Appended nucleophiles undergo 1,4-addition reactions with these intermediates to form cyclic vinyl oxazolidinones with good levels of diastereocontrol, highlighting a new approach to utilizing oxidative carbon-hydrogen bond functionalization to increase molecular complexity. PMID- 19552391 TI - Interfacial synthesis and widely controllable conductivity of polythiophene microparticles. AB - Fine polythiophene (PTh) microparticles were successfully synthesized by a novel interfacial polymerization at a dynamic interface between two immiscible solvents, i.e., n-hexane and acetonitrile or nitromethane containing thiophene and oxidant, respectively. The polymerization yield, size, and electrical conductivity of the microparticles are optimized by facilely regulating the medium species, oxidant species, oxidant/monomer ratio, monomer concentration, and polymerization temperature. The microparticles were thoroughly characterized by IR, UV-vis spectroscopy, wide-angle X-ray diffractometry, laser particle-size analyzer, and simultaneous TG-DSC technique. The yield rises with increasing oxidant/monomer ratio, monomer concentration, and polymerization temperature. However, low monomer concentration, low polymerization temperature, and modest oxidant/monomer ratio are all favorable for the formation of the PTh with good, large pi-conjugation and high conductivity. With decreasing the thiophene concentration from 200 to 50 mM at a fixed FeCl3/thiophene molar ratio of 3 at 0 degrees C in hexane/nitromethane biphase system, the PTh obtained exhibits a steadily enhanced conductivity from 10(-12) to 0.01 S cm(-1) and gradually darkening color from crimson to black. Under the same conditions, the PTh obtained in hexane/acetonitrile usually possesses lower yield but higher conductivity than that in hexane/nitromethane. The conductivity will be further enhanced to 1.1 and 4.4 S cm(-1) if the PTh powders are doped in iodine vapor and simply carbonized at 25 through 999 degrees C in nitrogen, respectively. The PTh is fine particles with the number-average diameter of 2.67-3.95 microm and low size polydispersity index between 1.12 and 1.23. The black particles carbonized at 25 to 999 degrees C are much smaller than original PTh particles, with the number-average diameter of 279 nm and size polydispersity index of 1.09. This interfacial approach provides an optimal synthesis of unique PTh microparticles with large pi-conjugation, high conductivity, black color, uniform size, good insolubility, excellent infusibility, high thermostability, and high yield of electrically conducting char at 999 degrees C. PMID- 19552392 TI - Calculation of free-energy differences by confinement simulations. Application to peptide conformers. AB - Conformational free-energy differences are key quantities for understanding important phenomena in molecular biology that involve large structural changes of macromolecules. In this paper, an improved version of the confinement approach, which is based on earlier developments, is used to determine the free energy of individual molecular states by progressively restraining the corresponding molecular structures to pure harmonic basins, whose absolute free energy can be computed by normal-mode analysis. The method is used to calculate the free-energy difference between two conformational states of the alanine dipeptide in vacuo, and of the beta-hairpin from protein G with an implicit solvation model. In all cases, the confinement results are in excellent agreement with the ones obtained from converged equilibrium molecular dynamics simulations, which have a much larger computational cost. The systematic and statistical errors of the results are evaluated and the origin of the errors is identified. The sensitivity of the calculated free-energy differences to structure-based definitions of the molecular states is discussed. A variant of the method, which closes the thermodynamic cycle by a quasi-harmonic rather than harmonic analysis, is introduced. The latter is proposed for possible use with explicit solvent simulations. PMID- 19552393 TI - Accurate relativistic small-core pseudopotentials for actinides. energy adjustment for uranium and first applications to uranium hydride. AB - The options to adjust accurate relativistic energy-consistent pseudopotentials for actinides are explored using uranium as an example. The choice of the reference data and the core-valence separation is discussed in view of a targeted accuracy of 0.04 eV or better in atomic energy differences such as excitation energies and ionization potentials. A new small-core pseudopotential attributing 60 electrons to the core has been generated by an energy adjustment to state averaged multiconfiguration Dirac-Hartree-Fock/Dirac-Coulomb-Breit Fermi nucleus reference data of 100 nonrelativistic configurations of U to U(7+) corresponding to 30190 reference J levels. At the finite-difference multiconfiguration Hartree Fock level the mean absolute errors are 0.002 and 0.024 eV for the configurations and J levels, respectively. A first molecular application to uranium monohydride UH yields very satisfactory agreement with results from all-electron calculations based on the Douglas-Kroll-Hess Hamiltonian. PMID- 19552394 TI - 12C/13C ratio in ethane on titan and implications for methane's replenishment. AB - The (12)C/(13)C abundance ratio in ethane in the atmosphere of Titan has been measured at 822 cm(-1) from high spectral resolution ground-based observations. The value, 89(8), coincides with the telluric standard and also agrees with the ratio seen in the outer planets. It is almost identical to the result for ethane on Titan found by the composite infrared spectrometer (CIRS) on Cassini. The (12)C/(13)C ratio for ethane is higher than the ratio measured in atmospheric methane by Cassini/Huygens GCMS, 82.3(1), representing an enrichment of (12)C in the ethane that might be explained by a kinetic isotope effect of approximately 1.1 in the formation of methyl radicals. If methane is being continuously resupplied to balance photochemical destruction, then we expect the isotopic composition in the ethane product to equilibrate at close to the same (12)C/(13)C ratio as that in the supply. The telluric value of the ratio in ethane then implies that the methane reservoir is primordial. PMID- 19552395 TI - Water's role in reshaping a macrocycle's binding pocket: infrared and ultraviolet spectroscopy of benzo-15-crown-5-(H(2)O)(n) and 4'-aminobenzo-15-crown-5 (H(2)O)(n), n = 1, 2. AB - Laser-induced fluorescence (LIF), resonant two-photon ionization (R2PI), ultraviolet hole-burning (UVHB), resonant ion-dip infrared (RIDIR), and infrared infrared ultraviolet hole-burning (IR-IR-UV) spectroscopies were carried out on benzo-15-crown-5 ether-(H(2)O)(n) (B15C-(H(2)O)(n)) and 4'-amino-benzo-15-crown-5 ether-(H(2)O)(n) (ABC-(H(2)O)(n)) clusters with n = 1,2 formed in a supersonic expansion. Two isomers of B15C-(H(2)O)(1) with S(0)-S(1) origins at 35,628 and 35,685 cm(-1) (B15C-(H(2)O)(1)(A) and B15C-(H(2)O)(1)(B), respectively) were identified and, on the basis of the combined evidence from the single-isomer UV and IR spectra, assigned to structures in which the H(2)O molecule donates both its OH groups to H-bonds to the crown oxygens. Both isomers share the same open, chairlike C(s) symmetry structure for the crown ether that exposes the crown oxygen lone pairs to binding to H(2)O on the interior of the crown. This crown conformation is not among those represented in the observed conformers in the absence of the H(2)O molecule, indicating that even a single water molecule is capable of reshaping the crown binding pocket in binding to it. In B15C (H(2)O)(1)(A), the water molecule takes up a position parallel to the crown plane of symmetry, using one OH group to bind to the two benzo oxygens, while the other OH binds to a single crown oxygen on the opposite side of the crown. The H(2)O molecule in B15C-(H(2)O)(1)(B) binds to the other two crown oxygens, in an orientation perpendicular to the crown's symmetry plane. B15C-(H(2)O)(2) also has two isomers. The first, B15C-(H(2)O)(2)(A) with S(0)-S(1) origin at 35,813 cm( 1), is assigned to a structure in which the two water molecules take up the two positions occupied by individual water molecules in B15C-(H(2)O)(1) A and B. The second isomer, with S(0)-S(1) origin at 35,665 cm(-1), has an OH stretch RIDIR spectrum that reflects a water-water H-bond, with the second water molecule binding to the crown-bound water in the parallel binding site. The combined data from B15C-(H(2)O)(1), ABC-(H(2)O)(1), and ABC-(HOD) complexes is used to deduce the uncoupled OH stretch wavenumber shifts associated with each of the unique binding sites for H(2)O to the crown. Arguments are presented that the binding pocket present in benzo-15-crown-5 ether is of a near ideal size to accommodate strong bidentate binding of individual water molecules to its most open crown conformation. PMID- 19552396 TI - Study of the benzocaine transfer from aqueous solution to the interior of a biological membrane. AB - The precise molecular mechanism of general anesthetics remains unknown. It is therefore important to understand where molecules with anesthetic properties localize within biological membranes. We have determined the free energy profile of a benzocaine molecule (BZC) across a biological membrane using molecular dynamics simulation. We use an asymmetric phospholipid bilayer with DPPS in one leaflet of a DPPC bilayer (Lopez Cascales et al. J. Phys. Chem. B 2006, 110, 2358 2363) to model a biological bilayer. From the free energy profile, we predict the zone of actuation of a benzocaine is located in the hydrocarbon region or at the end of the lipid head, depending of the presence of charged lipids (DPPS) in the leaflet. We observe a moderate increase in the disorder of the membrane and in particular an increase in the disorder of DPPS. Static and dynamic physicochemical properties of the benzocaine, such as its dipole orientation, translational diffusion coefficient, and rotational relaxation time were measured. PMID- 19552397 TI - Jet-cooled electronic and vibrational spectroscopy of crown ethers: benzo-15 crown-5 ether and 4'-amino-benzo-15-crown-5 ether. AB - Laser-induced fluorescence (LIF), ultraviolet hole-burning (UVHB), and resonant ion-dip infrared (RIDIR) spectroscopies were carried out on isolated benzo-15 crown-5 ether (B15C) and 4'-amino-benzo-15-crown-5 ether (ABC) cooled in a supersonic expansion. Three conformational isomers of B15C and four of ABC were observed and spectroscopically characterized. Full optimizations and harmonic frequency calculations were undertaken for the full set of almost 1700 conformational minima identified in a molecular mechanics force field search. When compared with TDDFT predictions, the S(0)-S(1) origin positions serve as a useful diagnostic of the conformation of the crown ether near the phenyl ring responsible for the UV absorption and to the position of the NH(2) substituent. In-plane orientations for the beta carbons produce red-shifted S(0)-S(1) origins, while out-of-plane "buckling" produces substantial blue shifts of 600 cm(-1) or more. Comparison between the alkyl CH stretch spectra of B15C and ABC divide the spectra into common subgroups shared by the two molecules. The high-frequency CH stretch transitions (above 2930 cm(-1)) reflect the number of CH...O interactions, which in turn track in a general way the degree of buckling of the crown. On this basis, assignments of each of the observed conformational isomers to a class of structure can be made. All the observed structures have some degree of buckling to them, indicating that in the absence of a strong-binding partner, the crown folds in on itself to gain additional stabilization from weak dispersive and CH...O interactions. PMID- 19552398 TI - HIV-1 peptide vaccine candidates: selecting constrained V3 peptides with highest affinity to antibody 447-52D. AB - The V3 region of the envelope glycoprotein gp120 of the human immunodeficiency virus type 1 (HIV-1) is a potential target for an anti-HIV-1 vaccine. Peptides corresponding to V3 form three variations of a beta-hairpin conformation when bound to anti-V3 HIV-1 neutralizing antibodies. The conformation of a V3(IIIB) peptide bound to the 0.5beta antibody, generated against an X4 gp120, has been postulated to represent the V3 conformation of X4 viruses while the conformations of a V3(MN) and a V3(CONSENSUS) peptide bound to the 447-52D human monoclonal antibody were postulated to represent the R5A and R5B V3 conformations of R5 viruses, respectively. To constrain the conformation of synthetic V3 peptides to these X4, R5A, and R5B conformations, we formed disulfide bonds between Cys residues whose location in a peptide template representing the entire V3(CONSENSUS) epitope recognized by the broadly neutralizing 447-52D antibody was changed systematically. In a previous study [Mor, A., et al. (2009) Biochemistry 48, 3288-3303] we showed that these constrained peptides adopted conformations resembling the three antibody-bound V3 conformations according to the location of the disulfide bonds. Here we show that these constrained peptides, with the exception of peptides in which the disulfide bond flanks the GPGR segment, retain high-affinity binding to the 447-52D antibody. Compared with peptides designed to mimic the X4 conformation, peptides designed to mimic either the R5A or R5B conformation had higher affinity to 447-52D. It is possible that constrained peptides which mimic the R5A and R5B conformations of the V3 and retain high affinity binding to 447-52D are good candidates for eliciting a broad neutralizing antibody response similar to that of 447-52D. PMID- 19552400 TI - Palladium-catalyzed acylation of sp2 C-H bond: direct access to ketones from aldehydes. AB - A palladium-catalyzed direct access to ketones from aldehydes via C-H cleavage of arenes is described. The procedure utilizes air as a clean and free terminal oxidant. PMID- 19552399 TI - Formation of carotenoid neutral radicals in photosystem II. AB - beta-Carotene radicals produced in the hexagonal pores of the molecular sieve Cu(II)-MCM-41 were studied by ENDOR and visible/near-IR spectroscopies. ENDOR studies showed that neutral radicals of beta-carotene were produced in humid air under ambient fluorescent light. The maximum absorption wavelengths of the neutral radicals were measured and were additionally predicted by using time dependent density functional theory (TD-DFT) calculations. An absorption peak at 750 nm, assigned to the neutral radical with a proton loss from the 4(4') position of the beta-carotene radical cation in Cu(II)-MCM-41, was also observed in photosystem II (PS II) samples using near-IR spectroscopy after illumination at 20 K. This peak was previously unassigned in PS II samples. The intensity of the absorption peak at 750 nm relative to the absorption of chlorophyll radical cations and beta-carotene radical cations increased with increasing pH of the PS II sample, providing further evidence that the absorption peak is due to the deprotonation of the beta-carotene radical cation. Based on a consideration of possible proton acceptors that are adjacent to beta-carotene molecules in photosystem II, as modeled in the X-ray crystal structure of Guskov et al. Nat. Struct. Mol. Biol. 2009, 16, 334-342, an electron-transfer pathway from a beta carotene molecule with an adjacent proton acceptor to P680*+ is proposed. PMID- 19552401 TI - Targeting and insertion of the cholesterol-binding translocator protein into the outer mitochondrial membrane. AB - Translocator protein (18 kDa, TSPO), previously known as the peripheral-type benzodiazepine receptor, is an outer mitochondrial membrane (OMM) protein necessary for cholesterol import and steroid production. We reconstituted the mitochondrial targeting and insertion of TSPO into the OMM to analyze the signals and mechanisms required for this process. Initial studies indicated the formation of a mitochondrial 66 kDa complex through Blue Native-PAGE analysis. The formation of this complex was found to be dependent on the presence of ATP and the cytosolic chaperone Hsp90. Through mutational analysis we identified two areas necessary for TSPO targeting, import, and function: amino acids 103-108 (Schellman motif), which provide the necessary structural orientation for import, and the cholesterol-binding C-terminus required for insertion. Although the translocase of the outer mitochondrial membrane (TOM) complex proteins Tom22 and Tom40 were present in the OMM, the TOM complex did not interact with TSPO. In search of proteins involved in TSPO import, we analyzed complexes known to interact with TSPO by mass spectrometry. Formation of the 66 kDa complex was found to be dependent on an identified protein, Metaxin 1, for formation and TSPO import. The level of import of TSPO into steroidogenic cell mitochondria was increased following treatment of the cells with cAMP. These findings suggest that the initial targeting of TSPO to mitochondria is dependent upon the presence of cytosolic chaperones interacting with the import receptor Tom70. The C-terminus plays an important role in targeting TSPO to mitochondria, whereas its import into the OMM is dependent upon the presence of the Schellman motif. Final integration of TSPO into the OMM occurs via its interaction with Metaxin 1. Import of TSPO into steroidogenic cell mitochondria is regulated by cAMP. PMID- 19552403 TI - Peptide-targeted diagnostics and radiotherapeutics. AB - Radiotherapeutic drugs and medical imaging agents, although used for different purposes, both benefit from precise targeting. When systemically administered, either would be most useful if designed to find and bind only a tumor, single type of cell, or unique molecular assembly thereon. In this Account, we examine the use of small peptides, natural and synthetic, to create biochemically specific molecular imaging agents and radiotherapeutic pharmaceuticals, discussing three distinct examples. In one project, a small natural peptide known to target members of the bombesin family of receptors was chemically attached to a strong, versatile metal chelator, DO3A, through a series of small-molecule linkers. The linkers powerfully affected not only binding strength for the bombesin receptors, tissue distribution, and tumor uptake in vivo but also receptor subtype specificity. When the assembly is combined with an active metal ion for human trials, the versatility of the DO3A (dodecanetriacetate) chelate affords choices in selecting the metal ion for different purposes: lutetium for a combination radiotherapeutic and diagnostic agent, (177)Lu-AMBA, and gallium for a positron emission tomography (PET) imaging agent, (68)Ga-AMBA. We also created small (approximately 5-kDa) bivalent peptides, each composed of different chemically linked peptides derived from phage display. The monomer peptides bound to the same target protein, VEGF-R2, a primary target of vascular endothelial growth factor (VEGF), the angiogenesis-stimulating protein. Several families of the monomer peptides did not compete with one another for the binding site on VEGF-R2. Their combination into fully synthetic hetero-bivalent molecules yielded subnanomolar K(d) values and greater than 100-fold improvements over homo bivalent molecules. Biological activity was evident in the hetero-bivalents, whereas none or very little existed in homo-bivalents, monomers, and monomer mixtures. In ultrasound imaging, tiny bubbles (2 microm in diameter) filled with inert gas can be used as effective contrast agents. By coating the shell of such bubbles with the peptide TKPPR (a tuftsin antagonist), we created contrast agents that bound unexpectedly to cultured endothelial cells expressing angiogenesis targets; the binding was attributable to a previously unnoticed and powerful multivalency effect. TKPPR binds specifically to neuropilin-1 (NP-1), a VEGF co receptor, but only when multimerized is it avid. Tuftsin, a small peptide derived from immunoglobulin G (IgG) that binds to macrophages during inflammation, has been studied for over 30 years; the receptor has never been cloned. Our results led to new conclusions about tuftsin, NP-1, and the purpose, heretofore unknown, of exon 8 in VEGF, which appears to be involved in NP-1 binding. Our disparate projects demonstrate that small-peptide targeted molecules can be very versatile in drug discovery in combination with classical medicinal chemistry. In particular, multivalent interactions can lead to unpredictable and useful biochemical information, as well as new drug candidates. PMID- 19552402 TI - Glutamate racemase dimerization inhibits dynamic conformational flexibility and reduces catalytic rates. AB - Glutamate racemase (RacE) is a bacterial enzyme that converts l-glutamate to d glutamate, an essential precursor for peptidoglycan synthesis. In prior work, we have shown that both isoforms cocrystallize with d-glutamate as dimers, and the enzyme is in a closed conformation with limited access to the active site [May, M., et al. (2007) J. Mol. Biol. 371, 1219-1237]. The active site of RacE2 is especially restricted. We utilize several computational and experimental approaches to understand the overall conformational dynamics involved during catalysis when the ligand enters and the product exits the active site. Our steered molecular dynamics simulations and normal-mode analysis results indicate that the monomeric form of the enzyme is more flexible than the native dimeric form. These results suggest that the monomeric enzyme might be more active than the dimeric form. We thus generated site-specific mutations that disrupt dimerization and find that the mutants exhibit significantly higher catalytic rates in the d-Glu to l-Glu reaction direction than the native enzyme. Low resolution models restored from solution X-ray scattering studies correlate well with the first six normal modes of the dimeric form of the enzyme, obtained from NMA. Thus, along with the local active site residues, global domain motions appear to be implicated in the catalytically relevant structural dynamics of this enzyme and suggest that increased flexibility may accelerate catalysis. This is a novel observation that residues distant from the catalytic site restrain catalytic activity through formation of the dimer structure. PMID- 19552404 TI - Signatures of nonequilibrium solvation dynamics on multidimensional spectra. AB - Multidimensional electronic and vibrational spectroscopies have established themselves over the last decade as uniquely detailed probes of intramolecular structure and dynamics. However, these spectroscopies can also provide powerful tools for probing solute-solvent interactions and the solvation dynamics that they give rise to. To this end, it should be noted that multidimensional spectra can be expressed in terms of optical response functions that differ with respect to the chromophore's quantum state during the various time intervals separating light-matter interactions. The dynamics of the photoinactive degrees of freedom during those time intervals (that is, between pulses) is dictated by potential energy surfaces that depend on the corresponding state of the chromophore. One therefore expects the system to hop between potential surfaces in a manner dictated by the optical response functions. Thus, the corresponding spectra should reflect the system's dynamics during the resulting sequence of nonequilibrium solvation processes. However, the interpretation of multidimensional spectra is often based on the assumption that they reflect the equilibrium dynamics of the photoinactive degrees of freedom on the potential surface that corresponds to the chromophore's ground state. In this Account, we present a systematic analysis of the signature of nonequilibrium solvation dynamics on multidimensional spectra and the ability of various computational methods to capture it. The analysis is performed in the context of the following three model systems: (A) a two-state chromophore with shifted harmonic potential surfaces that differ in frequency, (B) a two-state atomic chromophore in an atomic liquid, and (C) the hydrogen stretch of a moderately strong hydrogen bonded complex in a dipolar liquid. The following computational methods are employed and compared: (1) exact quantum dynamics (model A only), (2) the semiclassical forward-backward initial value representation (FB-IVR) method (models A and B only), (3) the linearized semiclassical (LSC) method (all three models), and (4) the standard ground-state equilibrium dynamics approach (all three models). The results demonstrate how multidimensional spectra can be used to probe nonequilibrium solvation dynamics in real time and with an unprecedented level of detail. We also show that, unlike the standard method, the LSC and FB IVR methods can accurately capture the signature of solvation dynamics on the spectra. Our results also suggest that LSC and FB-IVR yield similar results in the presence of rapid dephasing, which is typical in complex condensed-phase systems. This observation gives credence to the use of the LSC method for modeling spectra in complex systems for which an exact or even FB-IVR-based calculation is prohibitively expensive. PMID- 19552405 TI - Use of a temporary "solubilizing" peptide tag for the Fmoc solid-phase synthesis of human insulin glargine via use of regioselective disulfide bond formation. AB - Solid-phase peptide synthesis has been refined to a stage where efficient preparation of long and complex peptides is now achievable. However, the postsynthesis handling of poorly soluble peptides often remains a significant hindrance to their purification and further use. Several synthetic schemes have been developed for the preparation of such peptides containing modifications to aid their solubility. However, these require the use of complex chemistry or yield non-native sequences. We describe a simple approach based on the use of penta-lysine "tags" that are linked to the C-terminus of the peptide of interest via a base-labile linker. After ready purification of the now freely solubilized peptide, the "tag" is removed by simple, brief base treatment giving the native sequence in much higher overall yield. The applicability of the method was demonstrated by the novel preparation of insulin glargine via solid-phase synthesis of each of the two chains--including the notoriously poorly soluble A chain--followed by their combination in solution via regioselective disulfide bond formation. At the conclusion of the chain combination, the solubilizing peptide tag was removed from the A-chain to provide synthetic human glargine in nearly 10% overall yield. This approach should facilitate the development of new insulin analogues as well as be widely applicable to the improved purification and acquisition of otherwise poorly soluble synthetic peptides. PMID- 19552407 TI - Accelerated multidimensional NMR data acquisition by varying the pulse sequence repetition time. AB - Proposed is a simple method for shortening the data acquisition time in multidimensional NMR experiments based on variation of the repetition time (VRT). The repetition time is modulated (usually shortened) in concert with incrementing one or all of the evolution periods. The shortening of the data acquisition time is a consequence of accelerated pulsing. Initially, the pulsing is slow, and it is progressively accelerated toward the outskirts of the evolution domains. Pulsing modulation affects the longitudinal magnetization available for detection, causing line broadening during data acquisition, in complete analogy to filtering during data processing. In VRT, the only difference is that the filter is a function of longitudinal relaxation time. However, most often the resulting broadening is masked by the limited size of the evolution domain, so the overall effect of VRT is a slight reduction of the signal-to-noise ratio. The method is easy to implement in most multidimensional experiments since it only replaces the constant repetition time by a suitable VRT function. It is shown that in a VRT-NOESY-HSQC 3D experiment in 10 mM (13)C natural-abundance sucrose, the spectrum quality is virtually unchanged even when the data acquisition time is halved. PMID- 19552406 TI - Evaluation of a novel Arg-Gly-Asp-conjugated alpha-melanocyte stimulating hormone hybrid peptide for potential melanoma therapy. AB - The purpose of this study was to determine whether Arg-Gly-Asp (RGD)-conjugated alpha-melanocyte stimulating hormone (alpha-MSH) hybrid peptide could be employed to target melanocortin-1 (MC1) receptor for potential melanoma therapy. METHODS: The RGD motif {cyclic(Arg-Gly-Asp-DTyr-Asp)} was coupled to [Cys(3,4,10), DPhe(7), Arg(11)]alpha-MSH(3-13) {(Arg(11))CCMSH} to generate RGD-Lys (Arg(11))CCMSH hybrid peptide. The MC1 receptor binding affinity of RGD-Lys (Arg(11))CCMSH was determined in B16/F1 melanoma cells. The internalization and efflux, melanoma targeting and pharmacokinetic properties and single photon emission computed tomography/CT (SPECT/CT) imaging of (99m)Tc-RGD-Lys (Arg(11))CCMSH were determined in B16/F1 melanoma cells and melanoma-bearing C57 mice. Clonogenic cytotoxic effect of RGD-Lys-(Arg(11))CCMSH was examined in B16/F1 melanoma cells. RESULTS: RGD-Lys-(Arg(11))CCMSH displayed 2.1 nM MC1 receptor binding affinity. (99m)Tc-RGD-Lys-(Arg(11))CCMSH showed rapid internalization and extended retention in B16/F1 cells. The cellular uptake of (99m)Tc-RGD-Lys-(Arg(11))CCMSH was MC1 receptor-mediated. (99m)Tc-RGD-Lys (Arg(11))CCMSH exhibited high tumor uptake (14.83 +/- 2.94% ID/g 2 h postinjection) and prolonged tumor retention (7.59 +/- 2.04% ID/g 24 h postinjection) in B16/F1 melanoma-bearing mice. Nontarget organ uptakes were generally low except for the kidneys. Whole-body clearance of (99m)Tc-RGD-Lys (Arg(11))CCMSH was rapid, with approximately 62% of the injected radioactivity cleared through the urinary system by 2 h postinjection. Flank melanoma tumors were clearly imaged by small animal SPECT/CT using (99m)Tc-RGD-Lys-(Arg(11))CCMSH as an imaging probe 2 h postinjection. Single treatment (3 h incubation) with 100 nM of RGD-Lys-(Arg(11))CCMSH significantly (p < 0.05) decreased the clonogenic survival of B16/F1 cells by 65% compared to the untreated control cells. CONCLUSION: Favorable melanoma targeting property of (99m)Tc-RGD-Lys (Arg(11))CCMSH and remarkable cytotoxic effect of RGD-Lys-(Arg(11))CCMSH in B16/F1 cells warranted the further evaluation of (188)Re-labeled alpha-MSH hybrid peptides as novel therapeutic peptides for melanoma treatment once the strategies of amino acid coinjection or structural modification of peptide sequence substantially reduce the renal uptake. PMID- 19552408 TI - Immunological response from an entirely carbohydrate antigen: design of synthetic vaccines based on Tn-PS A1 conjugates. AB - An entirely carbohydrate-based immunogen consisting of a zwitterionic polysaccharide (ZPS) PS A1 and the well-known tumor antigen Tn has been designed, synthesized, and studied for immunological effects. The PS A1 motif was included to act as an MHCII elicitor for a T-cell-dependent immune response with increased immunogenicity against tumor-associated carbohydrate antigens, providing an alternative to carrier proteins. Through the use of C57BL/6 mice, it has been shown that chemical modification of PS A1 does not alter the recognition sequence responsible for an MHCII-mediated, T-cell-dependent immune response. The Tn-PS A1 conjugate construct confers specificity toward the Tn antigen alone, and specific carbohydrate immunoglobulins, namely, IgG3, are generated from intraperitoneal immunizations with or without adjuvant. The properties of the vaccine candidate are attributed to a site-specific linking strategy that incurs significant incorporation of Tn antigen. PMID- 19552410 TI - Fluorescent lifetime trajectories of a single fluorophore reveal reaction intermediates during transcription initiation. AB - Single molecule (SM) techniques are relatively new additions to the field of biophysics that allow one to manipulate individual molecules and study their behavior. To make these studies more relevant to what actually happens in the cell, one needs to move beyond the studies of individual molecules in isolation and study many different molecules working in concert. This presents a technical challenge as most SM experiments measure only one observable as a function of time, whereas complex biomolecular systems require multidimensional SM analysis. Forster resonance energy transfer (FRET) is one of the most common single molecule approaches and can report on the real time distance changes. However, FRET requires two fluorophores which will ultimately limit the degree of multiplexing in future SM applications. It will be useful if a single fluorophore can be used to provide equivalent information. In this communication, we show that fluorescence lifetime analysis of a single Cy3 fluorophore attached to the promoter region of the DNA can be used to reveal transient reaction intermediates during transcription initiation by T7 RNA polymerase. This work represents the first demonstration of real-time biochemical reactions observed via single molecule fluorescence lifetime trajectories of immobilized molecules. PMID- 19552409 TI - Modular nucleic acid assembled p/MHC microarrays for multiplexed sorting of antigen-specific T cells. AB - The human immune system consists of a large number of T cells capable of recognizing and responding to antigens derived from various sources. The development of peptide-major histocompatibility (p/MHC) tetrameric complexes has enabled the direct detection of these antigen-specific T cells. With the goal of increasing throughput and multiplexing of T cell detection, protein microarrays spotted with defined p/MHC complexes have been reported, but studies have been limited due to the inherent instability and reproducibility of arrays produced via conventional spotted methods. Herein, we report on a platform for the detection of antigen-specific T cells on glass substrates that offers significant advantages over existing surface-bound schemes. In this approach, called "Nucleic Acid Cell Sorting (NACS)", single-stranded DNA oligomers conjugated site specifically to p/MHC tetramers are employed to immobilize p/MHC tetramers via hybridization to a complementary-printed substrate. Fully assembled p/MHC arrays are used to detect and enumerate T cells captured from cellular suspensions, including primary human T cells collected from cancer patients. NACS arrays outperform conventional spotted arrays assessed in key criteria such as repeatability and homogeneity. The versatility of employing DNA sequences for cell sorting is exploited to enable the programmed, selective release of target populations of immobilized T cells with restriction endonucleases for downstream analysis. Because of the performance, facile and modular assembly of p/MHC tetramer arrays, NACS holds promise as a versatile platform for multiplexed T cell detection. PMID- 19552411 TI - Rational design of ligands targeting triplet repeating transcripts that cause RNA dominant disease: application to myotonic muscular dystrophy type 1 and spinocerebellar ataxia type 3. AB - Herein, we describe the design of high affinity ligands that bind expanded rCUG and rCAG repeat RNAs expressed in myotonic dystrophy type 1 (DM1) and spinocerebellar ataxia type 3. These ligands also inhibit, with nanomolar IC(50) values, the formation of RNA-protein complexes that are implicated in both disorders. The expanded rCUG and rCAG repeats form stable RNA hairpins with regularly repeating internal loops in the stem and have deleterious effects on cell function. The ligands that bind the repeats display a derivative of the bisbenzimidazole Hoechst 33258, which was identified by searching known RNA ligand interactions for ligands that bind the internal loop displayed in these hairpins. A series of 13 modularly assembled ligands with defined valencies and distances between ligand modules was synthesized to target multiple motifs in these RNAs simultaneously. The most avid binder, a pentamer, binds the rCUG repeat hairpin with a K(d) of 13 nM. When compared to a series of related RNAs, the pentamer binds to rCUG repeats with 4.4- to >200-fold specificity. Furthermore, the affinity of binding to rCUG repeats shows incremental gains with increasing valency, while the background binding to genomic DNA is correspondingly reduced. Then, it was determined whether the modularly assembled ligands inhibit the recognition of RNA repeats by Muscleblind-like 1 (MBNL1) protein, the expanded-rCUG binding protein whose sequestration leads to splicing defects in DM1. Among several compounds with nanomolar IC(50) values, the most potent inhibitor is the pentamer, which also inhibits the formation of rCAG repeat-MBNL1 complexes. Comparison of the binding data for the designed synthetic ligands and MBNL1 to repeating RNAs shows that the synthetic ligand is 23-fold higher affinity and more specific to DM1 RNAs than MBNL1. Further studies show that the designed ligands are cell permeable to mouse myoblasts. Thus, cell permeable ligands that bind repetitive RNAs have been designed that exhibit higher affinity and specificity for binding RNA than natural proteins. These studies suggest a general approach to targeting RNA, including those that cause RNA dominant disease. PMID- 19552412 TI - Two-dimensional electronic double-quantum coherence spectroscopy. AB - The theory of electronic structure of many-electron systems, such as molecules, is extraordinarily complicated. A consideration of how electron density is distributed on average in the average field of the other electrons in the system, that is, mean field theory, is very instructive. However, quantitatively describing chemical bonds, reactions, and spectroscopy requires consideration of the way that electrons avoid each other while moving; this is called electron correlation (or in physics, the many-body problem for fermions). Although great progress has been made in theory, there is a need for incisive experimental tests for large molecular systems in the condensed phase. In this Account, we report a two-dimensional (2D) optical coherent spectroscopy that correlates the double excited electronic states to constituent single-excited states. The technique, termed 2D double-quantum coherence spectroscopy (2D-DQCS), uses multiple, time ordered ultrashort coherent optical pulses to create double- and single-quantum coherences over time intervals between the pulses. The resulting 2D electronic spectrum is a map of the energy correlation between the first excited state and two-photon allowed double-quantum states. The underlying principle of the experiment is that when the energy of the double-quantum state, viewed in simple models as a double HOMO-to-LUMO (highest occupied to lowest unoccupied molecular orbital) excitation, equals twice that of a single excitation, then no signal is radiated. However, electron-electron interactions, a combination of exchange interactions and electron correlation, in real systems generates a signal that reveals precisely how the energy of the double-quantum resonance differs from twice the single-quantum resonance. The energy shift measured in this experiment reveals how the second excitation is perturbed by both the presence of the first excitation and the way that the other electrons in the system have responded to the presence of that first excitation. We compare a series of organic dye molecules and find that the energy offset for adding a second electronic excitation to the system relative to the first excitation is on the order of tens of millielectronvolts; it also depends quite sensitively on molecular geometry. These results demonstrate the effectiveness of 2D-DQCS for elucidating quantitative information about electron-electron interactions, many-electron wave functions, and electron correlation in electronic excited states and excitons. Our work helps illuminate the implications of electron correlation on chemical systems. In a broad sense, we are trying to help address the fundamental question "How do we go beyond the orbital representation of electrons in the chemical sciences?" PMID- 19552414 TI - Formation of two- and three-dimensional hybrid mesostructures from branched siloxane molecules. AB - We report the design of a new precursor having three branching disiloxane units capable of forming 3D mesostructures with a cubic Pm-3n and its orthorhombic and tetragonal variants Cmmm and P4(2)/mnm, in addition to a conventional 2D hexagonal (p6mm) mesostructure, thus creating a novel research area of mesostructural design in silica-organic nanohybrid materials. PMID- 19552413 TI - Palladium- and copper-catalyzed arylation of carbon-hydrogen bonds. AB - The transition-metal-catalyzed functionalization of C-H bonds is a powerful method for generating carbon-carbon bonds. Although significant advances to this field have been reported during the past decade, many challenges remain. First, most of the methods are substrate-specific and thus cannot be generalized. Second, conversions of unactivated (i.e., not benzylic or alpha to heteroatom) sp(3) C-H bonds to C-C bonds are rare, with most examples limited to t-butyl groups, a conversion that is inherently simple because there are no beta hydrogens that can be eliminated. Finally, the palladium, rhodium, and ruthenium catalysts routinely used for the conversion of C-H bonds to C-C bonds are expensive. Catalytically active metals that are cheaper and less exotic (e.g., copper, iron, and manganese) are rarely used. This Account describes our attempts to provide solutions to these three problems. We have developed a general method for directing-group-containing arene arylation by aryl iodides. Using palladium acetate as the catalyst, we arylated anilides, benzamides, benzoic acids, benzylamines, and 2-substituted pyridine derivatives under nearly identical conditions. We have also developed a method for the palladium-catalyzed auxiliary assisted arylation of unactivated sp(3) C-H bonds. This procedure allows for the beta-arylation of carboxylic acid derivatives and the gamma-arylation of amine derivatives. Furthermore, copper catalysis can be used to mediate the arylation of acidic arene C-H bonds (i.e., those with pK(a) values <35 in DMSO). Using a copper iodide catalyst in combination with a base and a phenanthroline ligand, we successfully arylated electron-rich and electron-deficient heterocycles and electron-poor arenes possessing at least two electron-withdrawing groups. The reaction exhibits unusual regioselectivity: arylation occurs at the most hindered position. This copper-catalyzed method supplements the well-known C-H activation/borylation methodology, in which functionalization usually occurs at the least hindered position. We also describe preliminary investigations to determine the mechanisms of these transformations. We anticipate that other transition metals, including iron, nickel, cobalt, and silver, will also be able to facilitate deprotonation/arylation reaction sequences. PMID- 19552416 TI - Alpha-hydroxyalkyl heterocycles via chiral allylic boronates: Pd-catalyzed borylation leading to a formal enantioselective isomerization of allylic ether and amine. AB - An efficient catalytic enantioselective preparation of synthetically useful pyranyl and piperidinyl allylic boronates was achieved via a palladium-catalyzed borylation/isomerization reaction on the corresponding alkenyl triflates. The influence of the base and solvent was found to be crucial on the regio- and enantioselectivity of this reaction. The overall borylation process constitutes a successful example of formal asymmetric isomerization of allylic ether/amine. The resulting allylic boronate reagents add to various aldehydes in a one-pot process to give synthetically useful alpha-hydroxyalkyl derivatives in high stereoselectivity. PMID- 19552418 TI - Mycobased synthesis of silver nanoparticles and their incorporation into sodium alginate films for vegetable and fruit preservation. AB - Biosynthesis of silver nanoparticles using Trichoderma viride and their incorporation into sodium alginate for vegetable and fruit preservation has been demonstrated in this study. Aqueous silver (Ag(+)) ions when exposed to the filtrate of T. viride are reduced in solution. These extremely stable silver nanoparticles were characterized by means of UV-vis spectrophotometer, FTIR, TEM, and EDS. The nanoparticles exhibit maximum absorbance at 421 nm in the UV spectrum. The presence of proteins was identified by FTIR. TEM micrograph revealed the formation of polydispersed nanoparticles, and the presence of elemental silver was confirmed by EDS analysis. The silver nanoparticle incorporated sodium alginate thin film shows good antibacterial activity against test strains. This film increases the shelf life of carrot and pear when compared to control with respect to weight loss and soluble protein content. These results show silver nanoparticle incorporated sodium alginate coated vegetables and fruits are suitable for preservation. PMID- 19552419 TI - Fluorescent-protein-based biosensors: modulation of energy transfer as a design principle. AB - Genetically-encoded biosensors based on FRET between fluorescent proteins of different hues enable quantitative measurement of intracellular enzyme activities and small molecule concentrations. (To listen to a podcast about this feature, please go to the Analytical Chemistry website at pubs.acs.org/journal/ancham.). PMID- 19552420 TI - Fourier transform infrared (FT-IR) microspectroscopic census of single starch granules for octenyl succinate ester modification. AB - Fourier transform infrared (FT-IR) microspectroscopy was used to investigate reaction homogeneity of octenyl succinic anhydride modification on waxy maize starch and detect uniformity of blends of modified and native starches. For the first time, the level and uniformity of chemical substitution on individual starch granules were analyzed by FT-IR microspectroscopy. More than 100 starch granules of each sample were analyzed one by one by FT-IR microspectroscopy. In comparison to the native starch, modified starch had two additional bands at 1723 and 1563 cm(-1), indicative of ester formation in the modified starch. For the 3% modification level, the degree of substitution (DS) was low (0.019) and the distribution of the ester group was not uniform among starch granules. For the modified starch with DS of 0.073, 99% of individual starch granules had a large carbonyl band area, indicating that most granules were modified to a sufficient extent that the presence of their carbonyl ester classified them individually as being modified. However, the octenyl succinate concentration varied between granules, suggesting that the reaction was not uniform. When modified starch (DS = 0.073) was blended with native starch (3:7, w/w) to achieve a mixture with an average DS of 0.019, FT-IR microspectroscopy was able to detect heterogeneity of octenyl succinate in the blend and determine the ratio of the modified starch to the native starch granules. PMID- 19552421 TI - Organocatalytic asymmetric formal [3 + 2] cycloaddition with in situ-generated N carbamoyl nitrones. AB - A novel organocatalytic formal [3 + 2] cycloaddition reaction with in situ generation of N-carbamoyl nitrones is presented. For the first time, N-Boc- and N Cbz-protected isoxazolidines have been directly obtained as single diastereoisomers in generally high yields and enantiomeric excesses using mild reaction conditions and inexpensive, readily available Cinchona alkaloid quaternary ammonium salts as catalysts. Synthetic manipulations of the products provided highly valuable building blocks such as free isoxazolidines, a N-Boc-1,3 aminoalcohol, and a free delta-lactam. This report represents a pioneering work in the use of N-carbamoyl nitrones as electron-poor 1,3-dipoles and glutaconates as new dipolarophiles in asymmetric catalysis. PMID- 19552423 TI - Simultaneous decoupled detection of dopamine and oxygen using pyrolyzed carbon microarrays and fast-scan cyclic voltammetry. AB - Microfabricated structures utilizing pyrolyzed photoresist have been shown to be useful for monitoring electrochemical processes. These previous studies, however, were limited to constant-potential measurements and slow-scan voltammetry. The work described in this paper utilizes microfabrication processes to produce devices that enable multiple fast-scan cyclic voltammetry (FSCV) waveforms to be applied to different electrodes on a single substrate. This enabled the simultaneous, decoupled detection of dopamine and oxygen. In this paper we describe the fabrication process of these arrays and show that pyrolyzed photoresist electrodes possess surface chemistry and electrochemical properties comparable to PAN-type, T-650, carbon fiber microelectrodes using background subtracted FSCV. The functionality of the array is discussed in terms of the degree of cross talk in response to flow injections of physiologically relevant concentrations of dopamine and oxygen. Finally, other applications of pyrolyzed photoresist microelectrode arrays are shown, including spatially resolved detection of analytes and combining FSCV with amperometry for the detection of dopamine. PMID- 19552424 TI - A fast capillary electrophoresis method for separation and quantification of modified nucleosides in urinary samples. AB - Modified nucleosides are formed at the post-transcriptional stage by chemical modification of normal nucleosides within the ribonucleic acid (RNA). These modified nucleosides cannot be reutilized or further degraded, but they are excreted in the urine as intact molecules. The elevated levels of modified nucleosides in the urine samples have served as potential cancer biomarkers in many studies. Although different analytical techniques have been reported for determining nucleosides levels, they are practically difficult to use as a routine tool for early cancer screening. In this paper, a novel method was developed to separate and quantify 10 nucleosides--adenosine, cytidine, guanosine, uridine, inosine, xanthosine, pseudouridine, N(2)-methylguanosine, 1 methyladenosine, and N(2),N(2)-dimethylguanosine--in urine samples using capillary electrophoresis with an ultraviolet (UV) detector (abbreviated as CE UV) at a wavelength of 254 nm. A 50 microm (i.d.) x 38 cm (effective length) fused silica capillary was used for the separation, and a borate-phosphate buffer containing 25 mM cetyltrimethylammonium bromide (CTAB) at pH 9.50 was used as a background electrolyte. The separation was performed at 15 kV under reverse polarity and completed within 10 min. The linear range of the analytes was 5.0 500 micromol/L, and the limit of detection was <2.0 micromol/L. The effects of pH, buffer concentrations, CTAB concentration, and the operation voltages on the separation and quantification of the modified nucleosides were also investigated. The technique developed in this study is much simpler and faster, compared to previous studies, and can be used to quantify modified nucleosides in urine samples. PMID- 19552425 TI - Native lignin structure of Miscanthus x giganteus and its changes during acetic and formic acid fractionation. AB - Milled wood lignin (MWL) and acetic and formic acid lignin (AL and FL) from Miscanthus x giganteus bark were produced, respectively, before and after organosolv fractionations under optimal conditions, in terms of organic and hydrochloric acid concentrations, liquid/wood ratio, and reaction time. In order to study the M. x giganteus native lignin structure and its modifications during the fractionation process, the lignins were studied by two-dimensional heteronuclear single quantum coherence (2D-(HSQC)), (13)C- and (31)P nuclear magnetic resonance (NMR) spectroscopy, Fourier transform infrared (FTIR), size exclusion chromatography (SEC) both before and after thioacidolysis, and elemental analysis. In addition, chemical composition analysis was performed on ash, Klason lignin, and carbohydrate content. The analyses demonstrated that M. x giganteus native lignin (MWL) is highly acylated at the C(gamma) of the lignin side chain (46%), possibly with p-coumarate and/or acetate groups. This is newsworthy since several earlier studies showed that acylation at the gamma carbon commonly occurs in C(3) and CAM grasses, whereas M. x giganteus is a C(4) grass. Furthermore, M. x giganteus showed a low S/G ratio (0.7) and a predominance of beta-O-4' linkages (up to 93% of all linkages). AL and FL lose part of these linkages during organosolv fractionation (up to 21 and 32%, respectively). The p-coumarate groups resist fractionation processes and are still present in high quantities in AL and FL. During the fractionation process, lignin is acetylated (acetic acid process) and condensed, with the G units condensing more than S units. M. x giganteus MWL contains a high content of carbohydrates (22.8%), suggesting that it is a lignin-carbohydrate complex (LCC). AL and FL showed low carbohydrate contents because of the breaking down of the LCC structures. AL and FL have high molecular weights and low polydispersities, and are high in phenolic content, qualities that make these suitable for different applications. These results suggest that refinement of M. x giganteus via organosolv processes could potentially turn this grass into a valuable source of both fiber and lignin. PMID- 19552426 TI - Functional group tolerant Kumada-Corriu-Tamao coupling of nonactivated alkyl halides with aryl and heteroaryl nucleophiles: catalysis by a nickel pincer complex permits the coupling of functionalized Grignard reagents. AB - A nickel(II) pincer complex [((Me)NN(2))NiCl] (1) catalyzes Kumada-Corriu-Tamao cross coupling of nonactivated alkyl halides with aryl and heteroaryl Grignard reagents. The coupling of octyl bromide with phenylmagnesium chloride was used as a test reaction. Using 3 mol % of 1 as the precatalyst and THF as the solvent, and in the presence of a catalytic amount of TMEDA, the coupling product was obtained in a high yield. The reaction conditions could be applied to cross coupling of other primary and secondary alkyl bromides and iodides. The coupling is tolerant to a wide range of functional groups. Therefore, alkyl halides containing ester, amide, ether, thioether, alcohol, pyrrole, indole, furan, nitrile, conjugated enone, and aryl halide moieties were coupled to give high isolated yields of products in which these units stay intact. For the coupling of ester-containing substrates, O-TMEDA is a better additive than TMEDA. The reaction protocol proves to be efficient for the coupling of Knochel-type functionalized Grignard reagents. Thus aryl Grignard reagents containing electron deficient and/or sensitive ester, nitrile, amide, and CF(3) substituents could be successfully coupled to nonactivated and functionalized alkyl iodides. The catalysis is also efficient for the coupling of alkyl iodides with functionalized heteroaryl Grignard reagents, giving rise to pyridine-, thiophene-, pyrazole-, furan-containing molecules with additional functionalities. Concerning the mechanism of the catalysis, [((Me)NN(2))Ni-(hetero)Ar] was identified as an intermediate, and the activation of alkyl halides was found to take place through a radical-rebound process. PMID- 19552428 TI - Vitamin B12 is the active corrinoid produced in cultivated white button mushrooms (Agaricus bisporus). AB - Analysis of vitamin B(12) in freshly harvested white button mushrooms ( Agaricus bisporus ) from five farms was performed by affinity chromatography and HPLC-ESI MS techniques. The vitamin B(12) concentrations obtained varied from farm to farm, with higher concentrations of vitamin B(12) detected in outer peel than in cap, stalk, or flesh, suggesting that the vitamin B(12) is probably bacteria derived. High concentrations of vitamin B(12) were also detected in the flush mushrooms including cups and flats. HPLC and mass spectrometry showed vitamin B(12) retention time and mass spectra identical to those of the standard vitamin B(12) and those of food products including beef, beef liver, salmon, egg, and milk but not of the pseudovitamin B(12), an inactive corrinoid in humans. The results suggest that the consumer may benefit from the consumption of mushroom to increase intake of this vitamin in the diet. PMID- 19552429 TI - A beta-galactosidase from pea seeds (PsBGAL): purification, stabilization, catalytic energetics, conformational heterogeneity, and its significance. AB - A basic glycosylated beta-galactosidase (PsBGAL) has been purified from pea seeds by 910-fold with a specific activity of 77.33 mumoL min(-1) mg(-1) protein. The purified enzyme is an electrophoretically homogeneous protein consisting of a single protein band with an apparent M(r) of 55 kDa, while the deglycosylated enzyme has a M(r) of 54.2 kDa on SDS-PAGE under reducing conditions. According to MALDI-TOF measurements of the 55 kDa band, the enzyme showed a homology with BGAL from other sources present in the SWISS-PROT database, while it showed no resemblance to any lectin. The N-terminal sequence of PsBGAL was determined as TIECK and showed a resemblance to BGAL from Arabidopsis thaliana (Q93Z24). The enzyme showed an unique property of multiple banding patterns on SDS-PAGE at 20 mA current, with tryptic digests of all bands having similar m/z values (using MALDI-TOF) while it showed only a single band at 10 mA current. PsBGAL is effectively compartmentalized during seed maturation inside vacuoles (pH approximately 5). The enzyme is capable of hydrolyzing pea seed xyloglucan, and it may be involved in modifying the cell wall architecture during seedling growth and development. The enzyme has a protonated carboxyl group at its active site as observed by ionization constant, thermodynamics, and chemical modification studies. PMID- 19552427 TI - Quantum mechanical/molecular mechanical and density functional theory studies of a prototypical zinc peptidase (carboxypeptidase A) suggest a general acid-general base mechanism. AB - Carboxypeptidase A is a zinc-containing enzyme that cleaves the C-terminal residue in a polypeptide substrate. Despite much experimental work, there is still a significant controversy concerning its catalytic mechanism. In this study, the carboxypeptidase A-catalyzed hydrolysis of the hippuryl-L-Phe molecule (k(cat) = 17.7 +/- 0.7 s(-1)) is investigated using both density functional theory and a hybrid quantum mechanical/molecular mechanical approach. The enzymatic reaction was found to proceed via a promoted-water pathway with Glu270 serving as the general base and general acid. Free-energy calculations indicate that the first nucleophilic addition step is rate-limiting, with a barrier of 17.9 kcal/mol. Besides activating the zinc-bound water nucleophile, the zinc cofactor also serves as an electrophilic catalyst that stabilizes the substrate carbonyl oxygen during the formation of the tetrahedral intermediate. In the Michaelis complex, Arg127, rather than Zn(II), is responsible for the polarization of the substrate carbonyl and it also serves as the oxyanion hole. As a result, its mutation leads to a higher free-energy barrier, in agreement with experimental observations. PMID- 19552430 TI - Anticancer activity of rhamnoallosan against DU-145 cells is kinetically complementary to coexisting Polyphenolics in Psidium guajava budding leaves. AB - Psidium guajava L. is a valuable farm fruit plant having many medicinal uses. Previously its budding leaves (PE) were shown to contain huge amounts of soluble polyphenolics (SP) including (in mg/g) gallic acid (348), catechin (102), epicatechin (60), rutin (100), quercetin (102), and rutin (100) and to exhibit potent anticancer activity. However, reconstitution of these polyphenolics recovered only 40% of the original bioactivity, and the soluble carbohydrate (SC) portion in PE was suspected to contribute the remaining. PE contained a novel rhamnoallosan, which had a carbohydrate/protein (w/w) ratio = 29.06%/10.27% (=2.83, average molecular mass of 5029 kDa), characteristically evidencing a peptidoglycan, consisting of a composition (mole % ratio) of rhamnose/allose/arabinose/tallose/xylose/fucose/glucose/mannose/galactose = 36.05:24.24:8.76:7.95:7.37:5.90:3.69:3.19:2.85 and of amino acid (in wt %) glycine/leucine/proline/alanine/methionine/isoleucine/valine/histidine/tyrosine/p enylalanine/cysteine/aspartic acid/lysine/glutamic acid = 37.12:12.68:10.05:8.97:5.99:4.89:4.83:4.25:4.05:2.78:1.86:1.10:0.73:0.70. Kinetic analysis showed comparable apparent cell-killing rate coefficients (k(app)) to be 4.03 x 10(3) and 2.92 x 10(3) cells mg(-1) h(-1), respectively, by SP and SC, evidencing the complementary anti-DU-145 bioactivity in nature. PMID- 19552431 TI - Novel small molecule bradykinin B2 receptor antagonists. AB - Blockade of the bradykinin B(2) receptor provides therapeutic benefit in hereditary angioedema (HAE) and potentially in many other diseases. Herein, we describe the development of highly potent B(2) receptor antagonists with a molecular weight of approximately 500 g/mol. First, known quinoline-based B(2) receptor antagonists were stripped down to their shared core motif 53, which turned out to be the minimum pharmacophore. Targeted modifications of 53 resulted in the highly water-soluble lead compound 8a. Extensive exploration of its structure-activity relationship resulted in a series of highly potent B(2) receptor antagonists, featuring a hydrogen bond accepting functionality, which presumably interacts with the side chain of Asn-107 of the B(2) receptor. Optimization of the microsomal stability and cytochrome P450 inhibition eventually led to the discovery of the highly potent and orally available B(2) receptor antagonist 52e (JSM10292), which showed the best overall properties. PMID- 19552432 TI - Octahydropyrrolo[3,4-c]pyrrole: a diamine scaffold for construction of either alpha4beta2 or alpha7-selective nicotinic acetylcholine receptor (nAChR) ligands. Substitutions that switch subtype selectivity. AB - A series of 5-(pyridine-3-yl)octahydropyrrolo[3,4-c]pyrroles have been prepared that exhibit high affinity to alpha4beta2 and/or alpha7 nicotinic acetylcholine receptors (nAChRs). Simple substitution patterns have been identified that allow construction of ligands that are highly selective for either nAChR subtype. The effects of substitution on subtype selectivity provide some insight into the differences in the ligand binding domains of the alpha4beta2 and alpha7 receptors, especially in regions removed from the cation binding pocket. PMID- 19552433 TI - Discovery of novel 2-aminobenzamide inhibitors of heat shock protein 90 as potent, selective and orally active antitumor agents. AB - A novel class of heat shock protein 90 (Hsp90) inhibitors was developed from an unbiased screen to identify protein targets for a diverse compound library. These indol-4-one and indazol-4-one derived 2-aminobenzamides showed strong binding affinity to Hsp90, and optimized analogues exhibited nanomolar antiproliferative activity across multiple cancer cell lines. Heat shock protein 70 (Hsp70) induction and specific client protein degradation in cells on treatment with the inhibitors supported Hsp90 inhibition as the mechanism of action. Computational chemistry and X-ray crystallographic analysis of selected member compounds clearly defined the protein-inhibitor interaction and assisted the design of analogues. 4-[6,6-Dimethyl-4-oxo-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H indazol-1-yl]-2-[(trans-4-hydroxycyclohexyl)amino]benzamide (SNX-2112, 9) was identified as highly selective and potent (IC(50) Her2 = 11 nM, HT-29 = 3 nM); its prodrug amino-acetic acid 4-[2-carbamoyl-5-(6,6-dimethyl-4-oxo-3 trifluoromethyl-4,5,6,7-tetrahydro-indazol-1-yl)-phenylamino]-cyclohexyl ester methanesulfonate (SNX-5422, 10) was orally bioavailable and efficacious in a broad range of xenograft tumor models (e.g. 67% growth delay in a HT-29 model) and is now in multiple phase I clinical trials. PMID- 19552435 TI - Chiral recognition based on enantioselectively aggregation-induced emission. AB - Novel chiral AIE compounds bearing a tartaric acid group were synthesized. They selectively aggregated with one enantiomer of a number of chiral amines, such that one enantiomer led to strong fluorescence and another enantiomer showed no or only weak fluorescence. This was used for the quantitative analysis of enantiomeric composition. PMID- 19552436 TI - In vitro intrinsic clearance-based optimization of N3-phenylpyrazinones as corticotropin-releasing factor-1 (CRF1) receptor antagonists. AB - A series of pyrazinone-based heterocycles was identified as potent and orally active corticotropin-releasing factor-1 (CRF(1)) receptor antagonists. Selected compounds proved efficacious in an anxiety model in rats; however, pharmacokinetic properties were not optimal. In this article, we describe an in vitro intrinsic clearance-based approach to the optimization of pyrazinone-based CRF(1) receptor antagonists wherein sites of metabolism were identified by incubation with human liver microsomes. It was found that the rate of metabolism could be decreased by incorporation of appropriate substituents at the primary sites of metabolism. This led to the discovery of compound 12x, a highly potent (IC(50) = 1.0 nM) and selective CRF(1) receptor antagonist with good oral bioavailability (F = 52%) in rats and efficacy in the defensive withdrawal anxiety test in rats. PMID- 19552438 TI - Probing the electronic stability of multiply charged anions: sulfonated pyrene tri- and tetraanions. AB - The strong intramolecular Coulomb repulsion in multiply charged anions (MCAs) creates a potential barrier that provides dynamic stability to MCAs and allows electronically metastable species to be observed. The 1-hydroxy-3,6,8-pyrene trisulfonate {[Py(OH)(SO(3))(3)](3-) or HPTS(3-)} was recently observed as a long lived metastable MCA with a large negative electron binding energy of -0.66 eV. Here we use Penning trap mass spectrometry to monitor the spontaneous decay of HPTS(3-) --> HPTS(*2-) + e(-) and have determined the half-life of HPTS(3-) to be 0.1 s. To explore the limit of electronic metastability, we tried to make the related quadruply charged pyrene-1,3,6,8-tetrasulfonate {[Py(SO(3))(4)](4-)}. However, only its decay product, the triply charged radical anion [Py(SO(3))(4)](*3-), as well as the triply charged ion-pairs [Py(SO(3))(4)H](3-) and [Py(SO(3))(4)Na](3-), was observed, suggesting that the tremendous intramolecular Coulomb repulsion makes the [Py(SO(3))(4)](4-) anion extremely short-lived. Photoelectron spectroscopy data showed that [Py(SO(3))(4)](*3-) is an electronically stable species with electron binding energies of +0.5 eV, whereas [Py(SO(3))(4)H](3-) and [Py(SO(3))(4)Na](3-) possess electron binding energies of 0.0 and -0.1 eV, respectively. Ab initio calculations confirmed the stability of these triply charged species and further predicted a large negative electron binding energy (-2.78 eV) for [Py(SO(3))(4)](4-), consistent with its short lifetime. PMID- 19552437 TI - Synthesis, structure-activity relationships, and in vivo evaluation of N3 phenylpyrazinones as novel corticotropin-releasing factor-1 (CRF1) receptor antagonists. AB - Evidence suggests that corticotropin-releasing factor-1 (CRF(1)) receptor antagonists may offer therapeutic potential for the treatment of diseases associated with elevated levels of CRF such as anxiety and depression. A pyrazinone-based chemotype of CRF(1) receptor antagonists was discovered. Structure-activity relationship studies led to the identification of numerous potent analogues including 12p, a highly potent and selective CRF(1) receptor antagonist with an IC(50) value of 0.26 nM. The pharmacokinetic properties of 12p were assessed in rats and Cynomolgus monkeys. Compound 12p was efficacious in the defensive withdrawal test (an animal model of anxiety) in rats. The synthesis, structure-activity relationships and in vivo properties of compounds within the pyrazinone chemotype are described. PMID- 19552439 TI - Ultrafast time-resolved infrared spectroscopy study of the photochemistry of N,N diethyldiazoacetamide: rearrangement in the excited state. AB - Ultrafast infrared spectroscopy shows that in chloroform, beta-lactam is formed immediately after the laser pulse but gamma-lactam is formed from both slow and fast processes. It is concluded that beta-lactam is formed from the diazoamide excited state via the rearrangement in the excited state (RIES) mechanism and that gamma-lactam is formed from both RIES and carbene. In methanol, both carbene decay and the rise of amide ether product are observed directly. Predictions from density functional theory calculations are consistent with these observations. PMID- 19552440 TI - Nature of molecular interactions of peptides with gold, palladium, and Pd-Au bimetal surfaces in aqueous solution. AB - We investigated molecular interactions involved in the selective binding of several short peptides derived from phage-display techniques (8-12 amino acids, excluding Cys) to surfaces of Au, Pd, and Pd-Au bimetal. The quantitative analysis of changes in energy and conformation upon adsorption on even {111} and {100} surfaces was carried out by molecular dynamics simulation using an efficient computational screening technique, including 1000 explicit water molecules and physically meaningful peptide concentrations at pH = 7. Changes in chain conformation from the solution to the adsorbed state over the course of multiple nanoseconds suggest that the peptides preferably interact with vacant sites of the face-centered cubic lattice above the metal surface. Residues that contribute to binding are in direct contact with the metal surfaces, and less binding residues are separated from the surface by one or two water layers. The strength of adsorption ranges from 0 to -100 kcal/(mol peptide) and scales with the surface energy of the metal (Pd surfaces are more attractive than Au surfaces), the affinity of individual residues versus the affinity of water, and conformation aspects, as well as polarization and charge transfer at the metal interface (only qualitatively considered here). A hexagonal spacing of approximately 1.6 A between available lattice sites on the {111} surfaces accounts for the characteristic adsorption of aromatic side groups and various other residues (including Tyr, Phe, Asp, His, Arg, Asn, Ser), and a quadratic spacing of approximately 2.8 A between available lattice sites on the {100} surface accounts for a significantly lower affinity to all peptides in favor of mobile water molecules. The combination of these factors suggests a "soft epitaxy" mechanism of binding. On a bimetallic Pd-Au {111} surface, binding patterns are similar, and the polarity of the bimetal junction can modify the binding energy by approximately 10 kcal/mol. The results are semiquantitatively supported by experimental measurements of the affinity of peptides and small molecules to metal surfaces as well as results from quantum-mechanical calculations on small peptide and surface fragments. Interfaces were modeled using the consistent valence force field extended for Lennard-Jones parameters for fcc metals which accurately reproduce surface and interface energies [Heinz, H.; Vaia, R. A.; Farmer, B. L.; Naik, R. R. J. Phys. Chem. C 2008, 112, 17281 17290]. PMID- 19552442 TI - Intramolecular hydroamination of aminoalkenes by calcium and magnesium complexes: a synthetic and mechanistic study. AB - The beta-diketiminate-stabilized calcium amide complex [{ArNC(Me)CHC(Me)NAr}Ca{N(SiMe(3))(2)}(THF)] (Ar = 2,6-diisopropylphenyl) and magnesium methyl complex [{ArNC(Me)CHC(Me)NAr}Mg(Me)(THF)] are reported as efficient precatalysts for hydroamination/cyclization of aminoalkenes. The reactions proceeded under mild conditions, allowing the synthesis of five-, six-, and seven-membered heterocyclic compounds. Qualitative assessment of these reactions revealed that the ease of catalytic turnover increases (i) for smaller ring sizes (5 > 6 > 7), (ii) substrates that benefit from favorable Thorpe-Ingold effects, and (iii) substrates that do not possess additional substitution on the alkene entity. Prochiral substrates may undergo diastereoselective hydroamination/cyclization depending upon the position of the existing stereocenter. Furthermore, a number of minor byproducts of these reactions, arising from competitive alkene isomerization reactions, were identified. A series of stoichiometric reactions between the precatalysts and primary amines provided an important model for catalyst initiation and suggested that these reactions are facile at room temperature, with the reaction of the calcium precatalyst with benzylamine proceeding with DeltaG(o)(298 K) = -2.7 kcal mol( 1). Both external amine/amide exchange and coordinated amine/amide exchange were observed in model complexes, and the data suggest that these processes occur via low-activation-energy pathways. As a result of the formation of potentially reactive byproducts such as hexamethyldisilazane, calcium-catalyst initiation is reversible, whereas for the magnesium precatalyst, this process is nonreversible. Further stoichiometric reactions of the two precatalysts with 1-amino-2,2 diphenyl-4-pentene demonstrated that the alkene insertion step proceeds via a highly reactive transient alkylmetal intermediate that readily reacts with N-H sigma bonds under catalytically relevant conditions. The results of deuterium labeling studies are consistent with the formation of a single transient alkyl complex for both the magnesium and calcium precatalysts. Kinetic analysis of the nonreversible magnesium system revealed that the reaction rate depends directly upon catalyst concentration and inversely upon substrate concentration, suggesting that substrate-inhibited alkene insertion is rate-determining. PMID- 19552441 TI - A highly reactive p450 model compound I. AB - The detection and kinetic characterization of a cytochrome P450 model compound I, [OFe(IV)-4-TMPyP](+) (1), in aqueous solution shows extraordinary reaction rates for C-H hydroxylations. Stopped-flow spectrophotometric monitoring of the oxidation of Fe(III)-4-TMPyP with mCPBA revealed the intermediate 1, which displays a weak, blue-shifted Soret band at 402 nm and an absorbance at 673 nm, typical of a porphyrin pi-radical cation. This intermediate was subsequently transformed into the well-characterized OFe(IV)-4-TMPyP. Global analysis afforded a second-order rate constant k(1) = (1.59 +/- 0.06) x 10(7) M(-1) s(-1) for the formation of 1 followed by a first-order decay with k(2) = 8.8 +/- 0.1 s(-1). (1)H and (13)C NMR determined 9-xanthydrol to be the major product (approximately 90% yield) of xanthene oxidation by 1. Electrospray ionization mass spectrometry carried out in 47.5% (18)OH(2) indicated 21% (18)O incorporation, consistent with an oxygen-rebound reaction scenario. Xanthene/xanthene-d(2) revealed a modest kinetic isotope effect, k(H)/k(D) = 2.1. Xanthene hydroxylation by 1 occurred with a very large second-order rate constant k(3) = (3.6 +/- 0.3) x 10(6) M(-1) s(-1). Similar reactions of fluorene-4-carboxylic acid and 4-isopropyl- and 4 ethylbenzoic acid also gave high rates for C-H hydroxylation that correlated well with the scissile C-H bond energy, indicating a homolytic hydrogen abstraction transition state. Mapping the observed rate constants for C-H bond cleavage onto the Bronsted-Evans-Polanyi relationship for similar substrates determined the H OFe(IV)-4-TMPyP bond dissociation energy to be approximately 100 kcal/mol. The high kinetic reactivity observed for 1 is suggested to result from a high porphyrin redox potential and spin-state-crossing phenomena. More generally, subtle charge modulation at the active site may result in high reactivity of a cytochrome P450 compound I. PMID- 19552445 TI - Copper-mediated direct arylation of 1,3,4-oxadiazoles and 1,2,4-triazoles with aryl iodides. AB - The copper-mediated direct arylation of 1,3,4-oxadiazoles and 1,2,4-triazoles with aryl iodides proceeds efficiently in the presence of suitable ligands and bases. This method allows the installation of a variety of aryl moieties bearing a functional group such as ketone, ester, or nitrile so as to enable the facile construction of various functionalized oxadiazole and triazole core pi systems. PMID- 19552444 TI - New S-adenosyl-L-methionine analogues: synthesis and reactivity studies. AB - Two new and complementary synthetic strategies for 5'-N-chloroethylamino-5' deoxyadenosines are presented. Additionally, the reaction kinetics of their conversion into aziridines under typical enzyme assay conditions is reported using time-resolved NMR spectroscopy. A stable photocaged derivative of 5'-N chloroethylamino-5'-deoxyadenosine has also been synthesized, and its stability and activation in aqueous solution at physiological pH have been examined. PMID- 19552446 TI - Lipase-catalyzed highly enantioselective kinetic resolution of boron-containing chiral alcohols. AB - The first application of enzymes as catalysts to obtain optically pure boron compounds is described. The kinetic resolution of boron-containing chiral alcohols via enantioselective transesterification catalyzed by lipases was studied. Aromatic, allylic, and aliphatic secondary alcohols containing a boronate ester or boronic acid group were resolved by lipase from Candida antartica (CALB), and excellent E values (E > 200) and high enantiomeric excesses (up to >99%) of both remaining substrates and acetylated product were obtained. PMID- 19552447 TI - Electron-transfer photoredox catalysis: development of a tin-free reductive dehalogenation reaction. AB - We report an operationally simple, tin-free reductive dehalogenation system utilizing the well-known visible-light-activated photoredox catalyst Ru(bpy)(3)Cl(2) in combination with (i)Pr(2)NEt and HCO(2)H or Hantzsch ester as the hydrogen atom donor. Activated C-X bonds may be reduced in good yields with excellent functional-group tolerance and chemoselectivity over aryl and vinyl C-X bonds. The proposed mechanism involves visible-light excitation of the catalyst, which is reduced by the tertiary amine to produce the single-electron reducing agent Ru(bpy)(3)(+). A subsequent single-electron transfer generates the alkyl radical, which is quenched by abstraction of a hydrogen atom. Reductions can be accomplished on a preparative scale with as little as 0.05 mol % Ru catalyst. PMID- 19552448 TI - Iron-catalyzed, hydrogen-mediated reductive cyclization of 1,6-enynes and diynes: evidence for bis(imino)pyridine ligand participation. AB - The bis(imino)pyridine iron dinitrogen complex (((i)Pr)PDI)Fe(N(2))(2) catalyzes the hydrogen-mediated reductive cyclization of enynes and diynes with turnover frequencies comparable to those of established precious metal catalysts. Amino, oxygenated, and carbon-based substrates are readily cyclized to the corresponding hetero- and carbocycles with 5 mol % iron and 4 atm H(2) at 23 degrees C. Stoichiometric reactions between selected substrates and the iron compound under a N(2) atmosphere established transfer dehydrogenation from an isopropyl aryl substituent to either the enyne or diyne substrate. In situ monitoring of the catalytic reaction by (1)H NMR spectroscopy coupled with deuterium labeling experiments established rapid cyclization followed by turnover-limiting hydrogenation. PMID- 19552449 TI - Iridium-catalyzed asymmetric hydrogenation yielding chiral diarylmethines with weakly coordinating or noncoordinating substituents. AB - Diarylmethine-containing stereocenters are present in pharmaceuticals and natural products, making the synthetic methods that form these chiral centers are important in industry. We have applied iridium complexes with novel N,P-chelating ligands to the asymmetric hydrogenation of trisubstituted olefins, forming diarylmethine chiral centers in high conversions and excellent enantioselectivities (up to 99% ee) for a broad range of substrates. Our results support the hypothesis that steric hindrance in one specific area of the catalyst is playing a key role in stereoselection, as the hydrogenation of substrates differing little at the prochiral carbon occurred with high enantioselectivity. As a result, excellent stereodiscrimination was obtained even when the prochiral carbon bore, for example, phenyl and p-tolyl groups. PMID- 19552450 TI - Heterobimetallic bismuth(III)/molybdenum(VI) and antimony(III)/molybdenum(VI) calix[5]arene complexes. Progress toward modeling the SOHIO catalyst. AB - The treatment of the monometallic bismuth or antimony complexes [M{(t)BuC5(H)(2)}] (M = Bi, Sb) with 1.5 equiv of MoO(2)(O(t)Bu)(2) in 1,2 dimethoxyethane (DME) produced soluble Bi(III)/Mo(VI) and Sb(III)/Mo(VI) heterometallic calix[5]arene complexes [Bi(2)Mo(4)O(11){(t)BuC5(H)}(2)] 1 and [Sb(2)Mo(4)O(11){(t)BuC5(H)}(2)] 2 in 55 and 45% yields, respectively. In solution the (1)H NMR patterns for 1 and 2 are characteristic of a C(s) symmetry with three pairs of doublets for the methylene protons and three singlets in a 1:2:2 ratio for the tert-butyl groups. Complex 1 crystallizes in the P1 space group and consists of a dimeric heterometallic Bi/Mo (1:2 ratio) complex featuring an overall Mo(4)Bi(2)O(21) oxo cluster. The remarkable oxo-rich core structure of 1 contains Bi(mu-O)Mo, Mo(mu-O)Mo, Mo=O...Mo, and Bi-O-Bi interactions that resemble aspects of the proposed SOHIO catalyst active site and the crystal structure of the Bi(2)Mo(2)O(9) oxide phase. PMID- 19552451 TI - The bologna annotation resource: a non hierarchical method for the functional and structural annotation of protein sequences relying on a comparative large-scale genome analysis. AB - Protein sequence annotation is a major challenge in the postgenomic era. Thanks to the availability of complete genomes and proteomes, protein annotation has recently taken invaluable advantage from cross-genome comparisons. In this work, we describe a new non hierarchical clustering procedure characterized by a stringent metric which ensures a reliable transfer of function between related proteins even in the case of multidomain and distantly related proteins. The method takes advantage of the comparative analysis of 599 completely sequenced genomes, both from prokaryotes and eukaryotes, and of a GO and PDB/SCOP mapping over the clusters. A statistical validation of our method demonstrates that our clustering technique captures the essential information shared between homologous and distantly related protein sequences. By this, uncharacterized proteins can be safely annotated by inheriting the annotation of the cluster. We validate our method by blindly annotating other 201 genomes and finally we develop BAR (the Bologna Annotation Resource), a prediction server for protein functional annotation based on a total of 800 genomes (publicly available at http://microserf.biocomp.unibo.it/bar/). PMID- 19552452 TI - Stabilization of imidosamarium(III) cubane by amidinates. AB - An imidosamarium(III) cubane has been prepared from the reductive cleavage of azobenzene by a divalent samarium bis(amidinate) complex, indicating that the "spectrator" bis(amidinate) and the resulting imido ligands help to stabilize the cubane framework. The cubane-type imido cluster is a novel unit in lanthanide chemistry. PMID- 19552453 TI - First-principles investigation of anistropic hole mobilities in organic semiconductors. AB - We report a simple first-principles-based simulation model (combining quantum mechanics with Marcus-Hush theory) that provides the quantitative structural relationships between angular resolution anisotropic hole mobility and molecular structures and packing. We validate that this model correctly predicts the anisotropic hole mobilities of ruberene, pentacene, tetracene, 5,11 dichlorotetracene (DCT), and hexathiapentacene (HTP), leading to results in good agreement with experiment. PMID- 19552454 TI - Investigation on phase separation kinetics of polyolefin blends through combination of viscoelasticity and morphology. AB - Phase separation kinetics of polyethylene copolymer blends polyethylene-co-hexene (PEH)/polyethylene-co-butene (PEB) at a phase separation temperature of 130 degrees C have been investigated through the combination of rheological measurements and optical microscope observation. When the blends are located in the unstable region, i.e., PEH/PEB 40/60 blend (H40), 50/50 blend (H50), and 60/40 blend (H60), due to the coeffect of the fast decay of concentration fluctuations and the reduced interfacial area, the stroage modulus, G', behaves dramatically, decreasing at the early or intermediate stages; while when the blends are located in the metastable region, i.e., PEH/PEB 70/30 blend (H70), G' decreases slightly and slowly during the whole time sweep process. During the cyclic frequency sweeps, G' evolutions of H50 and H70 show similar trends. Obviously different from the strong phase segregation systems, the increase of G' with time in the metastable region has not been observed, possibly due to the entanglement effects and weak interaction between the components of polyethylene blends. The interfacial tension-driven or diffusion-limited morphological evolutions of H50 and H70 during phase separation give direct interpretations to the viscoelastic difference between the two blends, which is dominated by different phase separation kinetics. The relatively low interfacial tensions at the late stage of phase separation for H50 (0.5-0.38 mN/m varying with time) and H70 (1.2 mN/m) can be estimated by using the Gramespacher-Meissner model. PMID- 19552455 TI - Surface adsorption and aggregate formation of cationic gemini surfactant and long chain alcohol mixtures. AB - We measured the surface tension of aqueous solutions of octanol-butandiyl-1,4 bis(decyldimethylammonium bromide) using the drop-volume technique at 298.15 K under atmospheric pressure as a function of the total molality and bulk composition. The results of the surface tension measurements, which were analyzed by originally developed thermodynamic equations, suggested that octanol molecules filled the spaces among the hydrophobic chains of gemini surfactants and formed a densely packed monolayer with them in the adsorbed film. The turbidity of aqueous solutions was also measured to construct the concentration-composition diagram with the surface tension data. A transmission electron microscope was used to determine the aggregate morphology in the aqueous solutions. Disc-like micelle and microemulsion regions were found on the diagram prior to the spherical micelle formation; nevertheless, the butandiyl-1,4-bis(decyldimethylammonium bromide) itself formed only spherical (or small ellipsoid) micelles in the concentration range measured. We also studied the relationship between synergism and molecular packing in the aggregates. PMID- 19552456 TI - Radiation effects on hydrophobic ionic liquid [C4mim][NTf2] during extraction of strontium ions. AB - The applications of room-temperature ionic liquids (RTILs) in separation of high level radioactive nuclides demand a comprehensive knowledge of the stability and metal ion extraction of RTILs under radiation. Herein, we assessed the influence of gamma-irradiation on the [C(4)mim][NTf(2)]-based extraction system, where [C(4)mim](+) is 1-butyl-3-methylimidazolium and [NTf(2)](-) is bis(trifluoromethylsulfonyl)imide, by solvent extraction of Sr(2+) using irradiated [C(4)mim][NTf(2)] in combination with dicyclohexyl-18-crown-6 (DCH18C6). It was found that the degree of extraction for Sr(2+) from water to irradiated [C(4)mim][NTf(2)] decreased compared with that to unirradiated [C(4)mim][NTf(2)], and the decrement enhanced obviously with increasing dose. NMR spectroscopic probe analysis revealed the formation of acids during irradiation of [C(4)mim][NTf(2)]. The decrease of Sr(2+) partitioning in irradiated [C(4)mim][NTf(2)] is attributed to the competition between H(+) with Sr(2+) to interact with DCH18C6. Accordingly, washing irradiated [C(4)mim][NTf(2)] with water gives a simple way of ionic liquid recycling. Furthermore, the degree of extraction for Sr(2+) from 3 mol.L(-1) nitric acid solution to [C(4)mim][NTf(2)] is independent of the irradiation of [C(4)mim][NTf(2)] since the amount of the radiation-generated H(+) is negligible in such a high acidic solution. PMID- 19552458 TI - Radiosynthesis of new [90Y]-DOTA-based maleimide reagents suitable for the prelabeling of thiol-bearing L-oligonucleotides and peptides. AB - We describe the radiosynthesis of two new [(90)Y]-DOTA-based maleimide reagents, suitable for the mild radiolabeling of L-RNAs and peptides modified with thiol bearing linkers. The synthesis procedure of both maleimide-bearing (90)Y complexes, [{(2S)-2-[4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)benzyl]-1,4,7,10 tetraazacyclododecane-1,4,7,10-tetrayl}tetraacetato][(90)Y]yttrate(1-)([(90)Y]3) and [{(2S)-2-(4-{[4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)butanoyl]amino}benzyl) 1,4,7,10-tetraaza-cyclododecane-1,4,7,10-tetrayl]tetraacetato}[(90)Y]yttrate(1 )([(90)Y]4), was optimized in terms of an easy purification method via solid phase extraction (SPE). Application as well as reactivity of both maleimide reagents were initially evaluated by the prelabeling of glutathione (GSH) and a thiol-modified 12mer L-RNA as model substances. In comparison to the N-aryl maleimide-bearing complex [(90)Y]3, N-alkyl maleimide-bearing complex [(90)Y]4 showed an increased hydrolytic stability at pH > or = 7. A slightly higher reactivity was found for [(90)Y]3 by prelabeling of 0.1 and 1 microg glutathione, respectively, in phosphate buffer (pH 7.2) at room temperature. In terms of very high radiochemical yields, the direct radiolabeling of DOTA-L-RNA conjugate with [(90)Y]YCl(3) proved to be more suitable than the prelabeling of the thiol modified 12mer L-RNA derivative with [(90)Y]4. PMID- 19552457 TI - Lipid-shelled vehicles: engineering for ultrasound molecular imaging and drug delivery. AB - Ultrasound pressure waves can map the location of lipid-stabilized gas micro bubbles after their intravenous administration in the body, facilitating an estimate of vascular density and microvascular flow rate. Microbubbles are currently approved by the Food and Drug Administration as ultrasound contrast agents for visualizing opacification of the left ventricle in echocardiography. However, the interaction of ultrasound waves with intravenously-injected lipid shelled particles, including both liposomes and microbubbles, is a far richer field. Particles can be designed for molecular imaging and loaded with drugs or genes; the mechanical and thermal properties of ultrasound can then effect localized drug release. In this Account, we provide an overview of the engineering of lipid-shelled microbubbles (typical diameter 1000-10 000 nm) and liposomes (typical diameter 65-120 nm) for ultrasound-based applications in molecular imaging and drug delivery. The chemistries of the shell and core can be optimized to enhance stability, circulation persistence, drug loading and release, targeting to and fusion with the cell membrane, and therapeutic biological effects. To assess the biodistribution and pharmacokinetics of these particles, we incorporated positron emission tomography (PET) radioisotopes on the shell. The radionuclide (18)F (half-life approximately 2 h) was covalently coupled to a dipalmitoyl lipid, followed by integration of the labeled lipid into the shell, facilitating short-term analysis of particle pharmacokinetics and metabolism of the lipid molecule. Alternately, labeling a formed particle with (64)Cu (half-life 12.7 h), after prior covalent incorporation of a copper chelating moiety onto the lipid shell, permits pharmacokinetic study of particles over several days. Stability and persistence in circulation of both liposomes and microbubbles are enhanced by long acyl chains and a poly(ethylene glycol) coating. Vascular targeting has been demonstrated with both nano- and microdiameter particles. Targeting affinity of the microbubble can be modulated by burying the ligand within a polymer brush layer; the application of ultrasound then reveals the ligand, enabling specific targeting of only the insonified region. Microbubbles and liposomes require different strategies for both drug loading and release. Microbubble loading is inhibited by the gas core and enhanced by layer-by-layer construction or conjugation of drug-entrapped particles to the surface. Liposome loading is typically internal and is enhanced by drug-specific loading techniques. Drug release from a microbubble results from the oscillation of the gas core diameter produced by the sound wave, whereas that from a liposome is enhanced by heat produced from the local absorption of acoustic energy within the tissue microenvironment. Biological effects induced by ultrasound, such as changes in cell membrane and vascular permeability, can enhance drug delivery. In particular, as microbubbles oscillate near a vessel wall, shock waves or liquid jets enhance drug transport. Mild heating induced by ultrasound, either before or after injection of the drug, facilitates the transport of liposomes from blood vessels to the tissue interstitium, thus increasing drug accumulation in the target region. Lipid-shelled vehicles offer many opportunities for chemists and engineers; ultrasound-based applications beyond the few currently in common use will undoubtedly soon multiply as molecular construction techniques are further refined. PMID- 19552459 TI - Synthesis and studies on cell-penetrating peptides. AB - The ability of different synthetic cell penetrating peptides, as Antennapedia (wild and Phe(6) mutated penetratins), flock house virus, and integrin peptides to form complexes with a 25mer antisense oligonucleotide was compared and their conformation was determined by circular dichroism spectroscopy. The efficiency for oligonucleotide delivery into cells was measured using peptides labeled with a coumarin derivative showing blue fluorescence and the fluorescein-labeled antisense oligonucleotide showing green fluorescence. Fluorescence due to the excitation energy transfer confirmed the interaction of the antisense oligonucleotide and cell-penetrating peptides. The most efficient oligonucleotide delivery was found for penetratins. Comparison of the two types of penetratins shows that the wild-type penetratin proved to be more efficient than mutated penetratin. The paper also emphasizes that the attachment of a fluorescent label may have an effect on the conformation and flexibility of cell-penetrating peptides that must be taken into consideration when evaluating biological experiments. PMID- 19552460 TI - Anion sensor-based ratiometric peptide probe for protein kinase activity. AB - A new fluorescent sensor consisting of Cd(II)-cylcen appended aminocoumarin and a substrate peptide for protein kinase A (PKA) has been designed. Upon phosphorylation by PKA, the metal complex moiety binds to a phosphorylated residue, which in turn displaces the coumarin fluorophore, and this event results in ratiometric change of excitation spectrum in neutral aqueous solution. PMID- 19552461 TI - Total synthesis of proximicin A-C and synthesis of new furan-based DNA binding agents. AB - The total synthesis of the natural occurring polyamides proximicin A-C (3-5) has been accomplished. A short and efficient synthesis of a thus far unknown 4-amino 2-furan carboxylic acid was developed. Furthermore, this unique heterocyclic gamma-amino-acid was used for the synthesis of a new class of AT-selective DNA binding agents derived from the natural products combining structural features of the proximicins with those from the known DNA-binding natural products netropsin (1) and distamycin (2). PMID- 19552462 TI - Efficient synthesis of regioisomerically pure bis(trifluoromethyl)-substituted 3,4,9,10-perylene tetracarboxylic bis(benzimidazole). AB - By a new strategy, two trifluoromethyl groups were efficiently introduced to highly insoluble n-type semiconductor 3,4,9,10-perylene tetracarboxylic bis(benzimidazole) (PTCBI), and for the first time we obtained three pure regioisomers of bis(trifluoromethyl)-substituted PTCBI, whose optical properties are different from one another. PMID- 19552463 TI - Pyrene-dihydrophenazine bis(radical cation) in a singlet ground state. AB - A new pyrene-dihydrophenazine dyad was prepared. Oxidation of the neutral species produced a bis(radical cation) species, which was characterized by the absorptions of their component radical cations in the visible region. A thermally accessible triplet state was observed in the ESR measurement in frozen n-PrCN. The energy gap between the singlet and triplet states was determined to be 2J/k(B) = -36 +/- 3 K. PMID- 19552464 TI - Design and synthesis of inhibitors of Hedgehog signaling based on the alkaloid cyclopamine. AB - The synthesis and biological evaluation of structurally simplified, metabolically stable cyclopamine-like Sonic Hedgehog (SHH) signaling inhibitors, i.e., 5, is described in four chemical steps from commercially available steroidal precursors. Biological evaluation of this cyclopamine analogue in two different systems establishes the high potency of 5 as a SHH signaling inhibitor. This approach provides important new lead structures for the development of new cancer chemotherapeutic agents based on the inhibition on SHH signaling. PMID- 19552466 TI - A novel synthesis of 1,3-oxazine-2,4-diones via a simple and efficient reaction of CO2 with 2,3-allenamides. AB - A simple and efficient reaction of CO(2) with 2,3-allenamides under mild conditions (CO(2) balloon without any metal catalyst in the presence of K(2)CO(3) or Cs(2)CO(3)) leads to an efficient synthesis of 1,3-oxazine-2,4-diones. The high reactivity of the allene moiety is crucial for the success of this transformation since the corresponding reaction of alpha,beta-unsaturated alkenamides or alkynamides does not occur. PMID- 19552465 TI - Syntheses and biological activity studies of novel sterol analogs from nitroso Diels-Alder reactions of ergosterol. AB - A series of novel sterol analogs was prepared using nitroso Diels-Alder reactions with ergosterol. Most cycloaddition reactions proceeded in an excellent regio- and stereoselective fashion. Further N-O bond cleavage of cycloadducts generated compounds with biological activity in PC-3 and MCF-7 cancer cell lines. PMID- 19552467 TI - Synthesis of 1,3-amino alcohol derivatives via a silicon-mediated ring-opening of substituted piperidines. AB - Multisubstituted piperidines containing a trimethylsilylmethyl group at C-2 can be opened regioselectively with TBAF and cyanogen bromide. The ring-opened products contain synthetically useful cyanamide and terminal alkene functional groups. This method is useful for the stereoselective synthesis of alkylamine derivatives containing multiple chiral centers. PMID- 19552470 TI - Autobiography of Robert Benny Gerber. PMID- 19552473 TI - Publications of Robert Benny Gerber. PMID- 19552468 TI - Direct, intermolecular, enantioselective, iridium-catalyzed allylation of carbamates to form carbamate-protected, branched allylic amines. AB - The direct reaction between carbamates and achiral allylic carbonates to form branched, conveniently protected primary allylic amines with high regioselectivity and enantioselectivity is reported. This process occurs without base or with 0.5 equiv K(3)PO(4) in the presence of a metalacyclic iridium catalyst containing a labile ethylene ligand. The reactions of aryl-, heteroaryl , and alkyl-substituted allylic carbonates with BocNH(2), FmocNH(2), CbzNH(2), TrocNH(2), TeocNH(2), and 2-oxazolidinone occur in good yields, with high selectivity for the branched isomer and high enantioselectivities (98% average ee). PMID- 19552474 TI - Statistics of indistinguishable particles. AB - The wave function of a system containing identical particles takes into account the relationship between a particle's intrinsic spin and its statistical property. Specifically, the exchange of two identical particles having odd-half integer spin results in the wave function changing sign, whereas the exchange of two identical particles having integer spin is accompanied by no such sign change. This is embodied in a term (-1)(2s), which has the value +1 for integer s (bosons), and -1 for odd-half-integer s (fermions), where s is the particle spin. All of this is well-known. In the nonrelativistic limit, a detailed consideration of the exchange of two identical particles shows that exchange is accompanied by a 2pi reorientation that yields the (-1)(2s) term. The same bookkeeping is applicable to the relativistic case described by the proper orthochronous Lorentz group, because any proper orthochronous Lorentz transformation can be expressed as the product of spatial rotations and a boost along the direction of motion. PMID- 19552475 TI - Photodissociation of H2(+) upon exposure to an intense pulsed photonic Fock state. AB - Producing and controlling nonclassical light states are now the subject of intense experimental efforts. In this paper we consider the interaction of such a light state with a small molecule. Specifically, we develop the theory and apply it numerically to calculate in detail how a short pulse of nonclassical light, such as the high intensity Fock state, induces photodissociation in H(2)(+). We compare the kinetic energy distributions and photodissociation yields with the analogous results of quasi-classical light, namely a coherent state. We find that Fock-state light decreases the overall probability of dissociation for low vibrational states of H(2)(+) as well as the location of peaks and line shapes in the kinetic energy distribution of the nuclei. PMID- 19552476 TI - Molecular dynamics simulations of bromine clathrate hydrates. AB - A polarizable force field that explicitly includes contributions from exchange repulsion, dispersion, charge penetration, and multipole electrostatics was developed to describe the interaction between bromine and water. This force field was combined with a polarizable force field for water and used in molecular dynamics simulations to calculate the relative energetics of three bromine clathrate hydrates. The simulations predict the tetragonal structure (Allen, K. W.; Jeffrey, G. A. J. Chem. Phys. 1963, 38, 2304) to be the most stable, with the CS-I and CS-II cubic structures being less stable. Although the CS-II species is not the most stable energetically, we argue that it could be formed under conditions of low bromine concentration, in agreement with recent measurements (Goldschleger, I. U.; Kerenskaya, G.; Janda, K. C.; Apkarian, V. A. J. Phys. Chem. A 2008, 112, 787) that provide evidence for three different bromine hydrate crystal types. PMID- 19552477 TI - Reduced-dimensional quantum approach to tunneling splittings using saddle-point normal coordinates. AB - We describe multidimensional extensions to a one-dimensional approach to tunneling splittings using a relaxed potential in the imaginary-frequency normal mode of the relevant saddle point (J. Chem. Phys. 2008 , 129 , 121103). Tests of these extensions are given for H(3)O(+) and NH(3) where full dimensional tunneling splittings are available and for the vinyl radical using a new full dimensional potential energy surface. PMID- 19552478 TI - Primary steps in the reaction of OH radicals with amino acids at low temperatures in Laval nozzle expansions: perspectives from experiment and theory. AB - Recent work has focused on the damaging effects of free radicals on biological molecules. This study investigates the kinetics of the attack of OH radicals on L alanine ethyl ester in the gas phase in cold beams of Laval nozzle expansions. Experiments and high-level theory are used to understand the preferred site of attack by the OH radical. Optimizations of L-alanine and L-alanine ethyl ester show that the essential transition state features for hydrogen abstraction off the C(alpha), C(beta), and N are similar. The energetics show that for L-alanine, the C(alpha)-site, C(beta)-site, and N-site transition states are all below the reactants level. For L-alanine ethyl ester, however, the energetics for hydrogen abstraction off the C(alpha) and N are the preferred site of reaction. These findings are supported by the observed negative temperature dependence of the rate constants of OH with alanine ethyl ester in Laval nozzle expansion experiments. More importantly, both the experiments and theory show that L alanine ethyl ester provides a good model for gas phase studies of the amino acids such as L-alanine. PMID- 19552479 TI - Coupled translation-rotation eigenstates of H2, HD, and D2 in the large cage of structure II clathrate hydrate: comparison with the small cage and rotational Raman spectroscopy. AB - We report fully coupled quantum five-dimensional calculations of the translation rotation (T-R) energy levels of one H(2), HD, and D(2) molecule confined inside the large hexakaidecahedral (5(12)6(4)) cage of the structure II clathrate hydrate. Highly converged T-R eigenstates have been obtained for excitation energies beyond the j = 2 rotational levels of the guest molecules, in order to allow comparison with the recent Raman spectroscopic measurements. The translationally excited T-R states are assigned with the quantum numbers n and l of the 3D isotropic harmonic oscillator. However, the translational excitations are not harmonic, since the level energies depend not only on n but also on l. For l > 1, the T-R levels having the same n,l values are split into groups of almost degenerate levels. The splitting patterns follow the predictions of group theory for the environment of T(d) symmetry, which is created by the configuration of the oxygen atoms of the large cage. The 2j + 1 degeneracy of the j = 1 and 2 rotational levels of the encapsulated hydrogen molecule is lifted entirely by the angular anisotropy of the H(2)-cage interaction potential. The patterns and magnitudes of the j = 1, 2 rotational level splittings, and the energies of the sublevels, in the large cage are virtually identical with those calculated for the small cage. This is in agreement with, and sheds light on, the observation that the S(0)(0) (j = 0-->2) bands in the rotational Raman spectra measured for simple H(2) hydrate and the binary hydrate of H(2) with tetrahydrofuran are remarkably similar with respect to their frequencies, widths, shapes, and internal structure, when the H(2) occupancy of the large cage of simple H(2) hydrate is low. PMID- 19552480 TI - Atomic spectral-product representations of molecular electronic structure: metric matrices and atomic-product composition of molecular eigenfunctions. AB - Recent progress is reported in development of ab initio computational methods for the electronic structures of molecules employing the many-electron eigenstates of constituent atoms in spectral-product forms. The approach provides a universal atomic-product description of the electronic structure of matter as an alternative to more commonly employed valence-bond- or molecular-orbital-based representations. The Hamiltonian matrix in this representation is seen to comprise a sum over atomic energies and a pairwise sum over Coulombic interaction terms that depend only on the separations of the individual atomic pairs. Overall electron antisymmetry can be enforced by unitary transformation when appropriate, rather than as a possibly encumbering or unnecessary global constraint. The matrix representative of the antisymmetrizer in the spectral-product basis, which is equivalent to the metric matrix of the corresponding explicitly antisymmetric basis, provides the required transformation to antisymmetric or linearly independent states after Hamiltonian evaluation. Particular attention is focused in the present report on properties of the metric matrix and on the atomic product compositions of molecular eigenstates as described in the spectral product representations. Illustrative calculations are reported for simple but prototypically important diatomic (H(2), CH) and triatomic (H(3), CH(2)) molecules employing algorithms and computer codes devised recently for this purpose. This particular implementation of the approach combines Slater-orbital based one- and two-electron integral evaluations, valence-bond constructions of standard tableau functions and matrices, and transformations to atomic eigenstate product representations. The calculated metric matrices and corresponding potential energy surfaces obtained in this way elucidate a number of aspects of the spectral-product development, including the nature of closure in the representation, the general redundancy or linear dependence of its explicitly antisymmetrized form, the convergence of the apparently disparate atomic-product and explicitly antisymmetrized atomic-product forms to a common invariant subspace, and the nature of a chemical bonding descriptor provided by the atomic product compositions of molecular eigenstates. Concluding remarks indicate additional studies in progress and the prognosis for performing atomic spectral product calculations more generally and efficiently. PMID- 19552481 TI - Delivery of polyethylenimine/DNA complexes assembled in a microfluidics device. AB - Polyethylenimine (PEI) and plasmid DNA (pDNA) complexes (PEI/pDNA) are nonviral vectors for gene delivery. The conventional method for producing these complexes involves bulk mixing (BM) of PEI and DNA followed by vortexing which at low N/P ratios results in large particle size distribution, low cytotoxicity, and poor gene transfection, while at high N/P ratios it results in small particle size and better gene transfection but high cytotoxicity. To improve size control, gene transfection efficiency, and cytotoxicity, in this study, we used a microfluidic hydrodynamic focusing (MF) device to prepare PEI/pDNA complexes at N/P = 3.3 and 6.7. We used bulk mixing as control, mouse NIH 3T3 fibroblast cells and mouse embryonic stem (mES) cells as model cell lines, plasmid encoding green fluorescent protein (pGFP) and secreted alkaline phosphatase (pSEAP) as the reporter gene, and commercially available Lipofectamine 2,000 as a positive control. The complexes were characterized by atomic force microscopy (AFM), dynamic light scattering (DLS), and zeta potential (zeta) measurement. Confocal laser scanning microscopy (CLSM) and fluorescent labeling techniques were used to visualize the complex size distribution, complexation uniformity, and cellular distribution. The results showed that MF produced complexes were smaller and more uniformly complexed and had higher cell viability and improved exogenous gene expression. PMID- 19552482 TI - Mapping of lysine monomethylation of linker histones in human breast and its cancer. AB - Linker histones H1 are key modulators of chromatin structure. Tightness of their binding to DNA is regulated by posttranslational modifications. In this study we have analyzed posttranslational modifications of five major variants of H1 in human tissue - H1.0, H1.2, H1.3, H1.4, and H1.5. To improve sequence coverage, tryptic peptides of H1 were separated by HPLC and the individual fractions were analyzed using a peptide on-chip implementation of nanoelectrospray (TriVersa), coupled to a linear ion trap-orbitrap hybrid instrument. For quantitative analysis of lysine methylation, ionization efficiencies of methylated and nonmethylated peptides were determined using synthetic peptides. Our analysis revealed that monomethylation of lysine residues alongside with phosphorylation of serine and threonine residues is the major modification of H1 in tissue. We found that most prominent methylation sites are in the N-terminal tail and the globular domain of H1. In the C- terminal domains we identified only few and less abundant methylation sites. Quantitative analysis revealed that up to 25% of H1.4 is methylated at K-26 in human tissues. Another prominent methylation site was mapped to K-27 in H1.5, which resembles the K-26 site in H1.4. In H1.0 five less abundant (<1% of H1.0) sites were identified. Analysis of patient matched pairs of cancer and adjacent normal breast demonstrated high variation in H1 methylation between individuals. PMID- 19552483 TI - Pharmacotherapy for cannabis dependence: how close are we? AB - Cannabis is the most widely used illicit drug in the world. Treatment admissions for cannabis use disorders have risen considerably in recent years, and the identification of medications that can be used to improve treatment outcomes among this population is a priority for researchers and clinicians. To date, several medications have been investigated for indications of clinically desirable effects among cannabis users (e.g. reduced withdrawal, attenuation of subjective or reinforcing effects, reduced relapse). Medications studied have included those: (i) known to be effective in the treatment of other drug use disorders; (ii) known to alleviate symptoms of cannabis withdrawal (e.g. dysphoric mood, irritability); or (iii) that directly affect endogenous cannabinoid receptor function. Results from controlled laboratory studies and small open-label clinical studies indicate that buspirone, dronabinol, fluoxetine, lithium and lofexidine may have therapeutic benefit for those seeking treatment for cannabis-related problems. However, controlled clinical trials have not been conducted and are needed to both confirm the potential clinical efficacy of these medications and to validate the laboratory models being used to study candidate medications. Although the recent increase in research towards the development of pharmacotherapy for cannabis use disorders has yielded promising leads, well controlled clinical trials are needed to support broad clinical use of these medications to treat cannabis use disorders. PMID- 19552484 TI - Lacosamide: a new approach to target voltage-gated sodium currents in epileptic disorders. AB - The mechanism of action of several antiepileptic drugs (AEDs) rests on their ability to modulate the activity of voltage-gated sodium currents that are responsible for fast action potential generation. Recent data indicate that lacosamide (a compound with analgesic and anticonvulsant effects in animal models) shares a similar mechanism. When compared with other AEDs, lacosamide has the unique ability to interact with sodium channel slow inactivation without affecting fast inactivation. This article reviews these findings and discusses their relevance within the context of neuronal activity seen during epileptiform discharges generated by limbic neuronal networks in the presence of chemical convulsants. These seizure-like events are characterized by sustained discharges of sodium-dependent action potentials supported by robust depolarizations, thus providing synchronization within neuronal networks. Generally, AEDs such as phenytoin, carbamazepine and lamotrigine block sodium channels when activated. In contrast, lacosamide facilitates slow inactivation of sodium channels both in terms of kinetics and voltage dependency. This effect may be relatively selective for repeatedly depolarized neurons, such as those participating in seizure activity in which the persistence of sodium currents is more pronounced and promotes neuronal excitation. The clinical effectiveness of lacosamide has been demonstrated in randomized, double-blind, parallel-group, placebo-controlled, adjunctive-therapy trials in patients with refractory partial seizures. Further studies should determine whether the effects of lacosamide in animal models and in clinical settings are fully explained by its selective action on sodium current slow inactivation or whether other effects (e.g. interactions with the collapsin-response mediator protein-2) play a contributory role. PMID- 19552485 TI - Protein kinase C inhibitors: rationale for use and potential in the treatment of bipolar disorder. AB - Bipolar disorder is one of the most severely debilitating of all medical illnesses. For a large number of patients, outcomes are quite poor. The illness results in tremendous suffering for patients and their families and commonly impairs functioning and workplace productivity. Risks of increased morbidity and mortality, unfortunately, are frequent occurrences as well. Until recently, little has been known about the specific molecular and cellular underpinnings of bipolar disorder. Such knowledge is crucial for the prospect of developing specific targeted therapies that are more effective and that have a more rapid onset of action than currently available treatments. Exciting recent data suggest that regulation of certain signalling pathways may be involved in the aetiology of bipolar disorder and that these pathways may be profitably targeted to treat the disorder. In particular, mania is associated with overactive protein kinase C (PKC) intracellular signalling, and recent genome-wide association studies of bipolar disorder have implicated an enzyme that reduces the activation of PKC. Importantly, the current mainstays in the treatment of mania, lithium (a monovalent cation) and valproate (a small fatty acid) indirectly inhibit PKC. In addition, recent clinical studies with the relatively selective PKC inhibitor tamoxifen add support to the relevance of the PKC target in bipolar disorder. Overall, a growing body of work both on a preclinical and clinical level indicates that PKC signalling may play an important role in the pathophysiology and treatment of bipolar disorder. The development of CNS-penetrant PKC inhibitors may have considerable benefit for this devastating illness. PMID- 19552488 TI - The efficacy and safety of blonanserin compared with haloperidol in acute-phase schizophrenia: a randomized, double-blind, placebo-controlled, multicentre study. AB - BACKGROUND: Blonanserin is a novel atypical antipsychotic agent with potent dopamine D(2) and serotonin 5-HT(2) antagonist properties. It may potentially have a lower incidence of adverse events than other antipsychotic agents. OBJECTIVE: To determine the efficacy and safety of three doses of blonanserin compared with placebo and haloperidol in patients with acute-phase schizophrenia. METHODS: This was a 6-week, randomized, double-blind, placebo- and haloperidol controlled, international, multicentre study. Patients with an acute exacerbation of their schizophrenia, with a Positive and Negative Syndrome Scale (PANSS) score >/=70 and a Clinical Global Impression - Severity of Illness (CGI-S) score >/=4 ('moderately ill') [with no decrease >/=20% or >/=1 point, respectively, during the wash-out period] were randomized into one of five treatment groups (blonanserin 2.5, 5 or 10 mg, haloperidol 10 mg or placebo once daily). Patients were assessed weekly for clinical efficacy, adverse events, extrapyramidal symptoms (EPS) and drug compliance, and were assessed biweekly for other safety variables. RESULTS: All 307 randomized patients received at least one dose of study medication and 228 (74.3%) completed the study. The mean reduction in PANSS total score at week 6 was significantly greater with all active treatments compared with placebo (-12.58; p < 0.001); blonanserin 10 mg was significantly superior to blonanserin 2.5 mg (-30.18 vs -20.6; p < 0.001), but blonanserin 5 mg (-27.19) and haloperidol 10 mg (-28.16) were not. All active treatments showed greater efficacy against the positive symptoms of schizophrenia, and blonanserin (5 and 10 mg) was more effective against the negative symptoms than haloperidol. Blonanserin was well tolerated at all doses and there was no evidence of clinically important weight gain, orthostatic hypotension, corrected QT interval prolongation or clinically relevant changes in laboratory test results. Haloperidol caused persistent elevation in prolactin levels, but this was not seen with any dose of blonanserin throughout the study period. There was a lower incidence of EPS with blonanserin 10 mg (26.6%) than with haloperidol 10 mg (53.3%). CONCLUSION: Blonanserin was effective in the treatment of acute schizophrenia and showed greater efficacy in negative symptoms compared with placebo and haloperidol. Blonanserin was well tolerated and its safety profile compared favourably with haloperidol, particularly with respect to prolactin elevation and EPS frequency. PMID- 19552486 TI - Cardiovascular abnormalities in patients with major depressive disorder: autonomic mechanisms and implications for treatment. AB - This article provides a detailed review of the association of major depression with coronary heart disease (CHD), examines the biological variables underpinning the linkage and discusses the clinical implications for treatment. When considering the co-morbidity between major depressive disorder (MDD) and CHD it is important to differentiate between (i) the prevalence and impact of MDD in those with existing CHD and (ii) MDD as a risk factor for the development of CHD. Whether the same biological mechanisms are at play in these two instances remains unknown. Depression is common in patients with CHD. Importantly, depression in these patients increases mortality. There is also consistent evidence that MDD is a risk factor for the development of CHD. The relative risk of developing CHD is proportional to the severity of depression and is independent of smoking, obesity, hypercholesterolaemia, diabetes mellitus and hypertension. There is a clear need to identify the underlying neurochemical mechanisms responsible for MDD and their linkage to the heart and vascular system. Of particular interest are activation of stress pathways, including both the sympathetic nervous system and hypothalamic-pituitary-adrenal axis, and inflammatory-mediated atherogenesis. Elevated sympathetic activity, reduced heart rate variability and increased plasma cortisol levels have been documented in patients with MDD. In addition to direct effects on the heart and vasculature, activation of stress pathways may also be associated with increased release of inflammatory cytokines such as interleukin-6 and tumour necrosis factor-alpha. Elevated levels of C-reactive protein are commonly observed in patients with MDD. The majority of investigations examining treatment of depression following myocardial infarction have focused on safety and efficacy; there is little evidence to indicate that treating depression in these patients improves survival. Given that strategies for preventive therapy remain incompletely formulated, future research should focus on generating a better understanding of the neurobiology of MDD and heart disease as a basis for rational and effective therapy. PMID- 19552487 TI - Cognitive effects of second-generation antipsychotics: current insights into neurochemical mechanisms. AB - Historically, pharmacotherapy for schizophrenia was mainly focused on finding drugs to treat psychotic symptoms only, without addressing other crucial domains of the disorder such as cognitive impairments. As a result, these domains have remained undertreated. In this review, we discuss recent preclinical research efforts, including investigation of synaptic mechanisms as well as intracellular signalling pathways and mechanisms involved in neuroplasticity and cell resilience, that may represent new mechanisms participating in the pathogenesis of schizophrenia, particularly at the level of the prefrontal cortex and hippocampus, and that might lead to the development of drugs that can counteract, at least partially, the cognitive impairments typical of schizophrenia. PMID- 19552490 TI - Antidepressants and falls in the elderly. AB - Antidepressants have long been recognized as a contributory factor to falls and many studies show an association between antidepressants and falls. There are extensive data for tricyclic antidepressants (TCAs) and related drugs, and for selective serotonin reuptake inhibitors (SSRIs), but few data for other classes of antidepressants. Sedation, insomnia and impaired sleep, nocturia, impaired postural reflexes and increased reaction times, orthostatic hypotension, cardiac rhythm and conduction disorders, and movement disorders have all been postulated as contributing factors to falls in patients taking antidepressants. Sleep disturbance is a cardinal feature of depression, and all antidepressants have effects on sleep. TCAs and related drugs cause marked sedation with daytime drowsiness. SSRIs and related drugs have an alerting effect, impairing sleep duration and quality and causing insomnia, which may result in nocturia and daytime drowsiness. Daytime drowsiness is a significant risk factor for falls, both in untreated depression and in depression treated with antidepressants. Clinically significant orthostatic hypotension is common with TCAs and related drugs, the older monoamine oxidase inhibitors and serotonin-norepinephrine reuptake inhibitors (SNRIs). It occurs less commonly with SSRIs, and rarely with moclobemide and bupropion, and is not reported as a significant adverse effect of hypericum (St John's wort). Cardiac rhythm and conduction disturbances are well recognized with TCAs, tetracyclics and SNRIs, but have also been reported with SSRIs. The contribution of antidepressant-induced conduction and rhythm disturbances to falls cannot be assessed with current data. There are insufficient data to exonerate any individual antidepressant or class of antidepressants as a potential cause of falls. The magnitude of the increased risk of falling with an antidepressant is about the same as the excess risk found in patients with untreated depression. PMID- 19552489 TI - Current treatment of pseudomonal infections in the elderly. AB - Pseudomonas aeruginosa infections have emerged as a major infectious disease threat in recent decades as a result of the significant mortality of pseudomonal pneumonia and bacteraemia, and the evolving resistance exhibited by the pathogen to numerous antibacterials. Pseudomonas possesses a large genome; thus, the pathogen is environmentally adaptable, metabolically flexible, able to overcome antibacterial pressure by selecting for resistant strains and even able to accumulate resistance mechanisms, leading to multidrug resistance (MDR), an increasingly recognized therapeutic challenge. In fact, most research currently does not focus on maximizing the efficacy of available antibacterials; rather, it focuses on maximizing their ecological safety. The elderly population may be particularly prone to pseudomonal infection as a result of increased co morbidities (such as diabetes mellitus and structural lung disease), the presence of invasive devices such as urinary catheters and feeding tubes, polypharmacy that includes antibacterials, and immune compromise related to age. However, age per se, as well as residence in nursing homes, may not predispose individuals to an increased risk for pseudomonal infection. On the other hand, age has been repeatedly outlined as a risk factor for MDR pseudomonal infections. The severity of pseudomonal infections necessitates prompt administration of appropriate antibacterials upon suspicion. Progress has been made in recognizing risk factors for P. aeruginosa infections both in hospitalized and community-residing patients. Antimicrobial therapy may be instituted as a combination or monotherapy: the debate cannot be definitively resolved since the available data are extracted from studies with varying targeted populations and varying definitions of response, adequacy and MDR. Empirical combination therapy maximizes the chances of bacterial coverage and exerts a lower resistance selection pressure. Although associated with increased percentages of adverse events, mainly as a result of the included aminoglycosides, empirical combination therapy seems a reasonable choice. Upon confirmation of Pseudomonas as the causative agent and awareness of its susceptibility profile, monotherapy is advocated by many, but not all, experts. Infections involving MDR strains can be treated with colistin, which has adequate efficacy and few renal adverse events, or doripenem. In the elderly, in addition to making dose modifications that are needed because of loss of renal function, the prescriber should be more cautious about the use of aminoglycoside-containing regimens, possibly replacing them with a combination of quinolone and a beta-lactam, notwithstanding the possible increased pressure for selection of resistance with the latter combination. PMID- 19552491 TI - Leflunomide treatment in elderly patients with rheumatoid or psoriatic arthritis: retrospective analysis of safety and adherence to treatment. AB - BACKGROUND: Disease-modifying antirheumatic drugs (DMARDs) play a crucial role in the treatment of persistent chronic synovitis, such as active rheumatoid arthritis (RA) and spondyloarthritis, by inducing or maintaining disease remission, reducing the frequency of flares or relapses, and allowing corticosteroids to be tapered while maintaining disease control. OBJECTIVE: The aim of this retrospective study was to evaluate the safety of, and adherence to treatment with, leflunomide in elderly RA and psoriatic arthritis patients compared with younger patients. METHODS: A total of 90 Italian patients (80 with active RA and 10 with psoriatic arthritis) were retrospectively examined at entry and after 24 months' follow-up. Patients were divided into two groups according to age: those aged 65 years (n = 40). Each patient was analysed for clinical, demographic and laboratory parameters in order to evaluate liver, renal and haematological toxicity. Disease Activity Score including a 28-joint count (DAS28) and physician global assessment of disease activity (MD global) were measured to define disease activity. RESULTS: During the 24-month follow-up period, 30 patients (33.3%) discontinued leflunomide: 17 patients (34.0%) in the group of patients aged 65 years. There were no differences in treatment withdrawal between the two groups. Overall, 10 patients (11.1%) in the entire study population discontinued leflunomide for lack of efficacy, while 21 (23.3%) discontinued the drug because of adverse effects (one patient withdrew because of both inefficacy and adverse effects). There were no significant differences in efficacy or adverse effects between patients aged 65 years. There was also no significant difference in survival rates of leflunomide treatment when patients aged 65 years (p = 0.94). There were no significant differences in withdrawal rates in the overall population when leflunomide monotherapy was compared with leflunomide combination therapy. There were also no significant differences in the types of adverse effects associated with monotherapy or combination therapy when the two age groups were compared. CONCLUSIONS: Leflunomide is a useful and well tolerated DMARD for the treatment of RA and psoriatic arthritis in the elderly. The safety profile of, and adherence to, leflunomide is not different in older patients with chronic inflammatory joint diseases such as RA or psoriatic arthritis to that observed in younger patients. PMID- 19552492 TI - Persistence with cholinesterase inhibitor therapy for dementia: an observational administrative health database study. AB - OBJECTIVE: To determine if choice of drug and ease of administration affect persistence of therapy with cholinesterase inhibitors (ChEIs) for treatment of dementia. METHODS: An observational administrative health database study was conducted in 5622 patients aged >or=65 years who received a new prescription for donepezil (DON), rivastigmine (RIV) or galantamine (GAL) from February to May 2006. Patients were followed for 1 year from initiation of therapy to determine percentage persistence and days of therapy. Once-daily galantamine extended release (GAL-ER) was compared with twice-daily galantamine immediate release (GAL IR) to determine if ease of administration affected persistence. Previous treatment with ChEIs was also documented. RESULTS: One-year persistence rates were significantly different among the ChEIs: GAL-ER 54% (95% CI 51, 57), DON 46% (95% CI 43, 49) and RIV 40% (95% CI 37, 43). Average days of therapy were greater for GAL-ER (293) than for RIV (272), but there were no differences between DON (287) and GAL-ER or DON and RIV. One-year persistence was significantly greater for GAL-ER 54% (95% CI 48, 59) than for GAL-IR 44% (95% CI 39, 50), although there was no significant difference in days of therapy (293 vs 286, respectively). More patients currently treated with RIV (40.5%) or GAL-ER (32.3%) had received previous treatment with a different ChEI than with DON (21.9%). CONCLUSION: Among possible factors affecting persistence of ChEI therapy for dementia, choice of drug, ease of administration and previous treatment appear to be important. PMID- 19552493 TI - Post-fracture prescribed calcium and vitamin D supplements alone or, in females, with concomitant anti-osteoporotic drugs is associated with lower mortality in elderly hip fracture patients: a prospective analysis. AB - BACKGROUND: Several studies have shown excess mortality among hip fracture patients compared with the normal population of the same age. Finnish guidelines for medical treatment of hip fracture patients recommend anti-osteoporosis medication and the daily concomitant use of prescribed calcium and vitamin D supplements. However, whether post-fracture use of calcium and vitamin D supplements is associated with survival in such patients has not been evaluated. OBJECTIVE: To study the association between survival in hip fracture patients and patients' sex and age, pre-fracture vitamin D status, American Society of Anesthesiologists - Physical Status (ASA-PS) class, type of fracture and post fracture use of prescribed calcium plus vitamin D and anti-osteoporotic medication. METHODS: The study population was 221 hip fracture patients primarily treated in acute care for a new hip fracture in 2003-4 in two Finnish hospitals. After a median of 27.5 months from the fracture, a questionnaire was sent to all patients who were still alive at the time (n = 137). The patients were queried about their use of prescribed calcium plus vitamin D supplementation and of anti osteoporotic drugs. The follow-up time for use of anti-osteoporotic medication and prescribed calcium and vitamin D was 19.5-36 months (median 27.5 months). Data on the use of prescribed calcium plus vitamin D supplementation and anti osteoporotic drugs were checked against information on reimbursement of drug prescriptions held by the Finnish Social Insurance Institution. A total of 4 years' (48 months') survival data for all patients in the study population was also obtained, with the dates of patient deaths being checked against Official National and Regional population statistics. Patient survival was analysed using both the Bayesian multivariate analysis and the life table method. RESULTS: In the multivariate analysis, the combination of variables that best explained post fracture survival was as follows: age <80 years; ASA-PS class 1-2 (ASA-PS class 1 and 2 data were combined in calculations); post-fracture use of prescribed calcium plus vitamin D supplements concomitantly with anti-osteoporotic drugs; post-fracture use of prescribed calcium plus vitamin D supplements; post-fracture use of anti-osteoporotic drugs only; and type of fracture (femoral neck or subtrochanteric). This model correctly predicted 74% of cases. At 36 months, we observed a 36% reduction in deaths in females who used prescribed calcium plus vitamin D supplementation and a corresponding 43% reduction in males. Survival of females who used anti-osteoporotic drugs concomitantly was even greater (43% reduction in deaths) over the entire follow-up period. Excess mortality was highest in females and males who used neither anti-osteoporotic drugs nor prescribed calcium and vitamin D. CONCLUSION: Our results indicate a potential relationship between reduced mortality and post-fracture use of prescribed calcium plus vitamin D supplementation and, in females, concomitant use of anti osteoporotic drugs. However, further investigations are needed to understand the reason for the reduction in the risk of death. Population-based, randomized, placebo-controlled trials with total mortality as the main endpoint should be conducted to verify our results. PMID- 19552495 TI - Brinzolamide/timolol: in open-angle glaucoma and ocular hypertension. AB - Brinzolamide 1%/timolol 0.5% fixed combination (brinzolamide/timolol) is a twice daily eyedrops suspension comprising the carbonic anhydrase-II inhibitor brinzolamide and the beta-adrenergic receptor antagonist timolol. Brinzolamide/timolol produced clinically relevant reductions in mean intraocular pressure (IOP) from baseline and was more effective than brinzolamide or timolol monotherapy in lowering IOP in a 6-month, randomized, phase III trial in patients with open-angle glaucoma or ocular hypertension (n = 523). The proportion of patients achieving a mean IOP of <18 mmHg was significantly greater in recipients of brinzolamide/timolol than in recipients of brinzolamide or timolol monotherapy. The IOP-lowering efficacy of brinzolamide/timolol was maintained for up to 12 months, and was no less effective than dorzolamide 2%/timolol 0.5% solution (dorzolamide/timolol) in a randomized, phase III, noninferiority trial (n = 437). Brinzolamide/timolol was generally well tolerated and was associated with significantly lower ocular discomfort scores than dorzolamide/timolol. Moreover, a significantly greater number of patients expressed a preference for brinzolamide/timolol over dorzolamide/timolol. The main ocular adverse event was blurred vision, and was not considered to be a safety issue. PMID- 19552494 TI - Prevalence of polypharmacy, polyherbacy, nutritional supplement use and potential product interactions among older adults living on the United States-Mexico border: a descriptive, questionnaire-based study. AB - BACKGROUND AND OBJECTIVE: The use of multiple medications, herbs or nutritional supplements can lead to adverse consequences, particularly in the elderly. A significant consequence resulting from polypharmacy, polyherbacy and nutritional supplement use is the potential for interactions to occur among the various products. The primary objective of this study was to estimate the prevalence of polypharmacy, polyherbacy, nutritional supplement use and potential product interactions among older adults living on the US-Mexico border. METHODS: This was a descriptive study that involved the administration of a bilingual (English/Spanish) questionnaire to a convenience sample of adults aged >or=60 years recruited from senior centres located within the most populated US-Mexico border region from June 2005 to March 2006. Participant demographics were collected in addition to information about current use of prescription medications, over-the-counter (OTC) medications, herbal products and nutritional supplements (i.e. nutraceuticals and vitamins or minerals). The outcomes measured were the number of prescription medications, OTC medications, herbal products, vitamins/minerals and nutraceuticals per participant. Furthermore, the number of potential interactions and major interactions between drugs, herbal products and nutritional supplements were identified for each participant. Additionally, product use patterns between men and women and among locations within the border region were compared. RESULTS: One-hundred-and-thirty participants (mean age 71.4 years) were recruited to complete the questionnaire. The prevalence of polypharmacy among all participants was 72.3% (n = 94), with 38.5% (n = 50) taking five or more concomitant medications (major polypharmacy). Twenty-one participants (16.2%) in the study sample reported taking two or more herbal products (polyherbacy). Thirty-four participants (26.2%) reported taking two or more vitamin/mineral supplements and nine (6.9%) reported using two or more nutraceuticals. Participants living on the US side of the border had higher rates of major polypharmacy, polyherbacy and use of nutritional supplements than those living on the Mexican side of the border. Overall, there were no significant differences in medication, herbal product and nutritional supplement use patterns between men and women. Evaluation of potential interactions revealed that 46.2% (n = 60) of participants were at risk of having at least one potential drug-drug interaction. Regarding drug and herbal product-supplement interactions, 31.5% (n = 41) of participants were at risk of having at least one possible interaction. CONCLUSIONS: The prevalence of polypharmacy among older adults living on the US side of the border was similar to national trends (estimates suggest that one quarter to one-half of US adults aged >or=65 years take five or more medications). However, polypharmacy was less common in older adults living on the Mexican side of the border. Additionally, herb use was higher in older adults living on the US-Mexico border than has been reported in national surveys of US adults, which indicate that less than one-quarter of adults have used a herbal product within the previous 12 months. Furthermore, this study demonstrated that older adults living on the US side of the border consumed more herbs and nutritional supplements than their Mexican counterparts. In addition to describing product use patterns on the border, these findings suggest that almost half of the older adult participants were at risk for a potential drug-drug interaction, with approximately one-third having a potential interaction between their medications, herbs or nutritional supplements. PMID- 19552497 TI - Revisiting bioaccumulation criteria for POPs and PBT assessments. AB - Scientists from academia, industry, and government reviewed current international regulations for the screening of commercial chemicals for bioaccumulation in the context of the current state of bioaccumulation science. On the basis of this review, several recommendations were proposed, including a scientific definition for "bioaccumulative substances," improved criteria for the characterization of bioaccumulative substances (including the trophic magnification factor and the biomagnification factor), novel methods for measuring and calculating bioaccumulation properties, and a framework for screening commercial chemicals for bioaccumulative substances. The proposed framework for bioaccumulation screening improves current practices by reducing miscategorization, making more effective use of available bioaccumulation data that currently cannot be considered, reducing the need for animal testing, providing simpler and cheaper test protocols for animal studies in case animal studies are necessary, making use of alternative testing strategies, including in vitro and in silico metabolic transformation assays, and providing a scientific foundation for bioaccumulation screening that can act to harmonize bioaccumulation screening among various jurisdictions. PMID- 19552498 TI - Environmental persistence of organic pollutants: guidance for development and review of POP risk profiles. AB - Environmental persistence is an important property that can enhance the potential of a chemical substance to exert adverse effects and be transported to remote environments. The persistence of organic compounds is governed by the rates at which they are removed by biological and chemical processes, such as biodegradation, hydrolysis, atmospheric oxidation, and photolysis. The persistence workgroup in a recent Society of Environmental Toxicology and Chemistry (SETAC) Pellston workshop (Pensacola, FL, USA, January 2008) focused on evaluating persistence of organic compounds in environmental media (air, water, soil, sediment) in terms of their single-medium degradation half-lives. The primary aim was to provide guidance to authors and reviewers of chemical dossiers for persistent organic pollutants (POPs) and persistent, bioaccumulative, and toxic substances (PBTs) proposed for action. A second objective was to provide a summary of the current state of the science with respect to POP fate assessment. Assessing the persistence of chemical substances in the environment is not straightforward. A common misconception is that, like many chemical properties, environmental persistence is an inherent property of the substance and can be readily measured. In fact, rates of degradation of a substance in the environment are determined by a combination of substance-specific properties and environmental conditions. This article addresses how persistence can be evaluated based on an assortment of supporting information. Special attention is given to several critical issues, including transformation products, nonextractable residues, and treatment of uncertainty and conflicting data as part of a weight of-evidence assessment. PMID- 19552499 TI - Use of measurement data in evaluating exposure of humans and wildlife to POPs/PBTs. AB - The Stockholm Convention on Persistent Organic Pollutants (POPs) recognized that POPs resist degradation, undergo long-range transport, and accumulate in remote ecosystems. The Stockholm Convention also acknowledged that indigenous communities, particularly in the Arctic, were at risk because of the biomagnification of POPs and contamination of their traditional foods. This recognition was largely based on environmental monitoring data and demonstrates the need to have adequate guidance on data collection and use. Although long range transport, persistence, and bioaccumulation models are important for screening potential POPs and for assessing human exposure, environmental measurement data are needed to confirm predictions. Indeed the Stockholm Convention (Annex E) requires monitoring data for assessing "exposure in local areas and, in particular, as a result of long-range environmental transport". However, there is relatively little guidance available on the most appropriate environmental measurement approaches, particularly for new candidate POPs, and on how to create a weight of evidence based on such data. We provide guidance on how to assess existing data that have been generated by monitoring programs and individual studies on the exposure of top predators and humans to candidate or potential POPs, as well as considerations for collecting new additional data. Our overall recommendation for assessing exposure in humans and top predators is to use or obtain direct measurements of the compound of concern from a significantly and uniquely exposed population (indigenous populations, remote populations), as well as data demonstrating biomagnification within food webs and time trends if possible. These data must be from the appropriate sample matrix type, collected and analyzed using accepted methodologies, reviewed for quality assurance, and interpreted correctly in order to be used to assess exposure. PMID- 19552500 TI - Evaluation of bioaccumulation using in vivo laboratory and field studies. AB - A primary consideration in the evaluation of chemicals is the potential for substances to be absorbed and retained in an organism's tissues (i.e., bioaccumulated) at concentrations sufficient to pose health concerns. Substances that exhibit properties that enable biomagnification in the food chain (i.e., amplification of tissue concentrations at successive trophic levels) are of particular concern due to the elevated long-term exposures these substances pose to higher trophic organisms, including humans. Historically, biomarkers of in vivo chemical exposure (e.g., eggshell thinning, bill deformities) retrospectively led to the identification of such compounds, which were later categorized as persistent organic pollutants. Today, multiple bioaccumulation metrics are available to quantitatively assess the bioaccumulation potential of new and existing chemicals and identify substances that, upon or before environmental release, may be characterized as persistent organic pollutants. This paper reviews the various in vivo measurement approaches that can be used to assess the bioaccumulation of chemicals in aquatic or terrestrial species using laboratory-exposed, field-deployed, or collected organisms. Important issues associated with laboratory measurements of bioaccumulation include appropriate test species selection, test chemical dosing methods, exposure duration, and chemical and statistical analyses. Measuring bioaccumulation at a particular field site requires consideration of which test species to use and whether to examine natural populations or to use field-deployed populations. Both laboratory and field methods also require reliable determination of chemical concentrations in exposure media of interest (i.e., water, sediment, food or prey, etc.), accumulated body residues, or both. The advantages and disadvantages of various laboratory and field bioaccumulation metrics for assessing biomagnification potential in aquatic or terrestrial food chains are discussed. Guidance is provided on how to consider the uncertainty in these metrics and develop a weight of-evidence evaluation that supports technically sound and consistent persistent organic pollutant and persistent, bioaccumulative, and toxic chemical identification. Based on the bioaccumulation information shared in 8 draft risk profiles submitted for review under the United Nations Stockholm Convention, recommendations are given for the information that is most critical to aid transparency and consistency in decision making. PMID- 19552501 TI - Use of (eco)toxicity data as screening criteria for the identification and classification of PBT/POP compounds. AB - Characterization of "significant adverse ecotoxicological effects" of persistent organic pollutants (POPs) presents particular challenges. In the various international conventions on POPs and persistent, bioaccumulative, and toxic substances, guidance on classification is not detailed and, in some cases, is unclear. This paper focuses on several key issues in relation to selection of assessment endpoints, use of appropriate effect measures, and uncertainty in the face of limited data. Because POPs are persistent and bioaccumulative, measures of effect should be based not on concentrations in environmental matrices but rather on residues in the body of the organism or in tissues that are causally linked to adverse responses. To obtain these data, currently used toxicity testing methods may need to be modified or substantiated by toxicokinetic information to ensure that substances with POP-like properties are adequately characterized. These data can be more easily matched to environmental monitoring measurements of body or tissue residues for the purposes of assessing whether adverse effects occur in the environment. In the face of persistence and accumulation in the food chain, and considering the extent and suitability of available data, a suitable policy on the use of uncertainty factors may need to be applied when making judgments about toxicity. This paper offers guidance that can be used to identify candidate POPs that have the potential to cause significant adverse effects in the ecosystem. PMID- 19552502 TI - Integrated approach to PBT and POP prioritization and risk assessment. AB - This article summarizes discussions at the SETAC Pellston Workshop on "Science Based Guidance and Framework for the Evaluation and Identification of PBTs and POPs" and provides an overview of other articles from that workshop that are also published in this issue. Identification of persistent, bioaccumulative, and toxic substances (PBTs) and persistent organic pollutants (POPs) and evaluation of their impact are more complicated than those for other chemicals and remain a challenge. The main reason for this is that PBT substance and POP assessment is associated with higher uncertainty and generally requires more data. However, for some data-rich PBTs and POPs, that identification and assessment of impact are feasible has been clearly demonstrated. New scientific developments and techniques are able to significantly increase the certainty of the various elements of PBT and POP assessment, and the current scientific literature provides many successful and illustrative examples that can be used as methodologies to build on. Applying multiple approaches for assessment is advisable, because it will reduce uncertainty and may increase confidence and improve the quality of decision-making. PMID- 19552503 TI - Modeling exposure to persistent chemicals in hazard and risk assessment. AB - Fate and exposure modeling has not, thus far, been explicitly used in the risk profile documents prepared for evaluating the significant adverse effect of candidate chemicals for either the Stockholm Convention or the Convention on Long Range Transboundary Air Pollution. However, we believe models have considerable potential to improve the risk profiles. Fate and exposure models are already used routinely in other similar regulatory applications to inform decisions, and they have been instrumental in building our current understanding of the fate of persistent organic pollutants (POP) and persistent, bioaccumulative, and toxic (PBT) chemicals in the environment. The goal of this publication is to motivate the use of fate and exposure models in preparing risk profiles in the POP assessment procedure by providing strategies for incorporating and using models. The ways that fate and exposure models can be used to improve and inform the development of risk profiles include 1) benchmarking the ratio of exposure and emissions of candidate chemicals to the same ratio for known POPs, thereby opening the possibility of combining this ratio with the relative emissions and relative toxicity to arrive at a measure of relative risk; 2) directly estimating the exposure of the environment, biota, and humans to provide information to complement measurements or where measurements are not available or are limited; 3) to identify the key processes and chemical or environmental parameters that determine the exposure, thereby allowing the effective prioritization of research or measurements to improve the risk profile; and 4) forecasting future time trends, including how quickly exposure levels in remote areas would respond to reductions in emissions. Currently there is no standardized consensus model for use in the risk profile context. Therefore, to choose the appropriate model the risk profile developer must evaluate how appropriate an existing model is for a specific setting and whether the assumptions and input data are relevant in the context of the application. It is possible to have confidence in the predictions of many of the existing models because of their fundamental physical and chemical, mechanistic underpinnings and the extensive work already done to compare model predictions and empirical observations. The working group recommends that modeling tools be applied for benchmarking PBT and POPs according to exposure-emissions relationships and that modeling tools be used to interpret emissions and monitoring data. The further development of models that combine fate, long-range transport, and bioaccumulation should be fostered, especially models that will allow time trends to be scientifically addressed in the risk profile. PMID- 19552504 TI - Multimedia partitioning, overall persistence, and long-range transport potential in the context of POPs and PBT chemical assessments. AB - For the identification and assessment of persistent, bioaccumulative, and toxic (PBT) chemicals and persistent organic pollutants (POPs), overall persistence (P(ov)) and long-range transport potential (LRTP) are important indicators. In this article, we first give an overview of methods to determine P(ov) and LRTP and discuss the influence of multimedia partitioning of semivolatile organic chemicals (SOCs) on P(ov) and LRTP. Next, we summarize the most important features of various multimedia fate and transport models that can be used to calculate P(ov) and LRTP. Complementary to environmental fate models, field data provide important empirical information about the spatial distribution and time trends of SOC concentrations in the environment. We discuss the role of field data in the estimation of P(ov) and LRTP and give an overview of important field studies showing the levels and trends of various groups of chemicals in different parts of the world. Then, we address key topics in the field of PBT and POP assessment that require further research, such as the formation of transformation products, the influence of atmospheric aerosols on the degradation and transport of SOCs, and the effect of long-range transport by ocean currents. In addition, we describe the most important types of uncertainty associated with estimates of P(ov) and LRTP, which are mainly uncertainty of chemical property data and uncertainty of the design of environmental fate models. Finally, we illustrate the characterization of SOCs in terms of P(ov) and LRTP with the example of the consensus model for P(ov) and LRTP Tool that is provided by the Organization for Economic Cooperation and Development. PMID- 19552505 TI - Introduction to special series: science-based guidance and framework for the evaluation and identification of PBTs and POPs. AB - There is a growing sense of urgency among scientists and environmental policy makers concerning the need for improving the scientific foundation supporting international regulations for identifying and evaluating persistent, bioaccumulative, and toxic (PBT) substances and persistent organic pollutants (POPs) in the environment. The current national and international regulations define PBTs and POPs in terms of fairly strict criteria that are based on the state of the science in the late 1970s and early 1980s. Since then, an evolution in the state of the science has produced new insights into PBT substances and an array of new methods to identify PBT chemicals. The development of regulatory criteria has not kept up with the rapid development in environmental chemistry and toxicology, and as a result, scientists often find themselves in the situation where guidance on PBT and POPs criteria is limited and, in some respects, out of date. With this background, a Society of Environmental Toxicology and Chemistry (SETAC) Pellston Workshop brought together experts from academia, government, and industry to reach consensus on the significance of advancements in our understanding of the behavior and potential impact of POPs and PBTs in the environment, the current understanding of the state of the science, as well as recommendations for policy-makers to improve and coordinate national and international regulations on this issue. The workshop builds on the outcome of a previous Pellston workshop, held in 1998, which focused on the evaluation of persistence and long-range transport of organic chemicals in the environment, and is linked to other recent Pellston workshops, among them the Tissue Residue Approach for Toxicity Assessment workshop held in 2007. The results of this workshop are conveyed in a series of 9 articles, published in this issue of Integrated Environmental Assessment and Management, and describe the coordination of science, regulation, and management needed to more effectively achieve a common goal of managing chemicals on our planet. PMID- 19552506 TI - Update on patented cholesterol absorption inhibitors. AB - BACKGROUND: Atherosclerosis is one of the most life-threatening diseases primarily associated with hypercholesterolemia and is characterized by increased serum cholesterol level. Cholesterol originates from both its de novo synthesis within the hepatic cells and its absorption into the intestine in the form of dietary or bile cholesterol. Interventions influencing both of these processes are promising therapeutic options to lower the cholesterol level. Hydroxymethyl glutaryl-CoA reductase inhibitors, commonly known as statins, effectively block the rate determining step in the biosynthesis of cholesterol. Ezetimibe is the first new class of drugs used to treat hypercholesterolemia by inhibition of cholesterol absorption through Niemann Pick C1 Like 1 membrane of enterocytes. Therefore, combination therapy of ezetimibe and statins offers an efficacious new approach for the prevention and treatment of hypercholesterolemia. OBJECTIVES: The present review focuses on updates on ezetimibe and patented profile of novel cholesterol absorption inhibitors followed by critical analysis of different targets such as cholesterol esterase inhibitors, bile acid transport inhibitors or phospholipase-A(2) inhibitors, etc.which play an important role in the lipid absorption. CONCLUSION: The discovery of ezetimibe has opened a new door for the management of hyper-cholesterolemia in combination with statins. There are newer analogues that are under clinical trials, among which darapladib, FM-VP4 and A 002 are promising compounds. PMID- 19552507 TI - A novel Syk kinase inhibitor suitable for inhalation: R-343(?)--WO-2009031011. AB - The subject of this application is N(4)-[(2,2-difluoro-4H-benzo[1,4]oxazin-3-one) 6-yl]-5-fluoro-N(2)-[3-(methylaminocarbonylmethyleneoxy)phenyl]-2,4 pyrimidinediamine xinafoate salt, as well as inhalant formulations of the salt and a process for the preparation of the salt. This salt is probably R-343, a Syk kinase inhibitor being developed under license by Pfizer, an inhaled formulation of which is in Phase I development for the treatment of asthma. PMID- 19552508 TI - Positive allosteric modulators of the metabotropic glutamate receptor 2 for the treatment of schizophrenia. AB - BACKGROUND: Normalization of excessive glutamate neurotransmission through activation of the metabotropic glutamate receptor 2 (mGluR2) represents a novel and promising approach for the treatment of schizophrenia. This strategy has gained support through the evaluation of dual mGluR2/3 agonists that act directly at the glutamate (orthosteric) binding site. Importantly, clinical validation of the mechanism was achieved in a Phase II study in schizophrenia patients with mGluR2/3 agonist LY404039. Selective positive allosteric modulators (potentiators) of mGluR2 that bind to the transmembrane region of the receptor have shown efficacy in rodent models predictive of antipsychotic activity, but have yet to be evaluated in the clinic. Allosteric mGluR2 potentiators may offer advantages over orthosteric mGluR2/3 agonists as a result of their unique mode of action and ability to achieve superior mGluR2 selectivity. OBJECTIVE/METHOD: This review focuses on the structures and biological activities of small molecule potentiators of mGluR2 that appeared in the patent literature between 2006 and early 2009. CONCLUSION: Potent mGluR2 potentiators that span a broad range of structural diversity have been disclosed. Narrow patent filings within select series and drug-like properties of corresponding preferred compounds suggest that development candidates have likely been nominated. PMID- 19552510 TI - Bradykinin receptor antagonists--a review of the patent literature 2005-2008. AB - BACKGROUND: For > 20 years, pharmaceutical companies and academic centers have been developing bradykinin antagonists. The patent literature on these molecules (up to and including 2004) has been analyzed previously in this journal in two review articles. OBJECTIVE: The aim of this review is to provide an update (from 2005 to early 2009) on the patenting activity in the field of bradykinin antagonists (including patents on their formulation). Where possible, the information from the patents has been supplemented with that from the primary literature, clinical trial databases and company websites in an attempt to give a more complete picture. CONCLUSIONS: In the past 4 years, nearly 50 new patents have been filed on bradykinin antagonists--in the case of several filings, only the original source has been considered in this analysis--the vast majority of these (> 93%) on B1 antagonists. However, despite this large amount of work, only one compound, icatibant--a hydrophilic decapeptide selective for the B2 receptor- has reached the market, although it needs to be administered parenterally. PMID- 19552509 TI - Flavin-containing monooxygenase 1-catalysed N,N-dimethylamphetamine N-oxidation. AB - N,N-dimethylamphetamine (DMA) is a methamphetamine analogue known to be a weaker central nervous system stimulant than methamphetamine. Although a major metabolite of DMA is known to be DMA N-oxide (DMANO), which may be catalysed by flavin-containing monooxygenase (FMO), the specific enzyme(s) involved in this biotransformation has not been identified. In this study, the specific enzyme(s) involved with DMA N-oxidation was characterized by several assays. When DMA was incubated with different human recombinant drug-metabolizing enzymes, including FMOs and cytochrome P450s (CYPs), the formation of DMANO by FMO1 was the most predominant. The Michaelis-Menten kinetic constants for DMA N-oxidation by FMO1 were: K(m) of 44.5 microM, V(max) of 7.59 nmol min(-1) mg(-1) protein, and intrinsic clearance of 171 microl min(-1) mg(-1) protein, which was about twelve fold higher than that by FMO3. Imipramine, an FMO1-specific inhibitor, selectively inhibited DMA N-oxidation. The resulting data showed that DMA N oxidation is mainly mediated by FMO1. PMID- 19552511 TI - Review of recent acetyl-CoA carboxylase inhibitor patents: mid-2007-2008. AB - BACKGROUND: Acetyl-CoA carboxylase (ACC) is a biologic target that is receiving increased attention for the treatment of obesity and type 2 diabetes mellitus. Inhibition of this enzyme, either in transgenic mice or pharmacologically, has been shown to have beneficial effects on lab animals. METHOD: This review of the ACC inhibitor patent literature covers the period from mid-2007 to December 2008, during which time a total of 18 patents were published. CONCLUSION: These published patent applications include ACC inhibitors that inhibit the enzyme through modulation of the carboxyltransferase-domain, inhibitors that bind to the biotin carboxylase-domain and novel chemotypes whose mode of action was not disclosed. Furthermore, published patents claim the discovery of ACC2 isoform selective and ACC1/2 non-selective inhibitors. PMID- 19552512 TI - Novel patent publications on high-affinity nicotinic acid receptor agonists. AB - BACKGROUND: Nicotinic acid (NA) has been used as a drug to treat dyslipidemia for > 50 years. In outcome clinical trials, NA displayed remarkable efficacy in patients with cardiovascular diseases by modifying lipid profiles that results in reduced morbidity and mortality. On the other hand, NA induces vasodilation (flushing) that undermines treatment compliance. In addition, high-dose treatment is required presumably owing to the poor pharmacokinetic properties of NA. The identification of the high-affinity NA receptor, namely G-protein coupled receptor 109A (GPR109A), led to further understanding of the pharmacological effects of NA and discovery of compounds that are potentially superior in efficacy yet devoid of NA's adverse effects. OBJECTIVE/METHOD: This review focuses on the endeavors of several pharmaceutical companies to discover and develop GPR109A agonists. Representative compounds of each series in patent literature since 2005 are highlighted. CONCLUSION: Highly potent GPR109A agonists with minimal flushing effects and robust free fatty acid reduction have been identified. Despite the failure of the partial agonist MK-0354 to achieve efficacy in a Phase II clinical trial, at least three other GPR109A agonists have been evaluated in clinical trials. The upcoming clinical data would be critical to validate the therapeutic utility of this receptor. PMID- 19552513 TI - Chemical penetration enhancers: a patent review. AB - BACKGROUND: Ever since transdermal drug delivery came into existence, it has offered great promises, although most of them are yet to be fulfilled owing to some intrinsic restrictions of the transdermal route. On the positive side, transdermal drug delivery systems present advantages including non-invasiveness, prolonged therapeutic effect, reduced side effects, improved bioavailability, better patient compliance and easy termination of drug therapy. The greatest hindrance in the percutaneous delivery is the obstruction property of the stratum corneum, the outermost layer of the skin, in addition to usual problems such as skin binding, skin metabolism, cutaneous toxicity and prolonged lag times. OBJECTIVE: This paper reviews investigations on the feasibility and application of penetration enhancers as described in recent patents, which help in the selection of a suitable sorption promoter(s) for enhanced delivery of medicaments through the skin. METHOD: The patents granted under various categories of penetration enhancers have been discussed including fatty acids, terpenes, fatty alcohol, pyrrolidone, sulfoxides, laurocapram, surface active agents, amides, amines, lecithin, polyols, quaternary ammonium compounds, silicones, alkanoates and so on. CONCLUSION: Scores of promising chemicals have been harnessed for their skin permeation promoting capacity as mentioned earlier. In future, many more chemicals and putative enhancers are likely be documented and patented. PMID- 19552516 TI - Humanized anti-EphB4 antibodies for the treatment of carcinomas and vasculogenesis-related diseases. AB - The invention provides human, humanized or chimeric versions of anti-EphB4 mouse monoclonal antibodies that bind to the human EphB4 receptor tyrosine kinase. The described anti-EphB4 antibodies are derived from two murine mAbs #47 and #131 through framework shuffling and include those of the IgGl, IgG2, IgG3 or IgG4 human isotype. The patent further relates to pharmaceutical compositions, immunotherapeutic compositions and methods using therapeutic antibodies that bind to the human EphB4 antigen and that may induce phosphorylation and degradation of EphB4 and mediate antigen-dependent cell-mediated-cytotoxicity, complement dependent cell-mediated cytotoxicity and/or apoptosis for the treatment of human malignancies and vasculogenesis-related disorders and diseases. PMID- 19552517 TI - Investigation and control of an outbreak of Enterobacter aerogenes bloodstream infection in a neonatal intensive care unit in Fiji. AB - Ten neonates developed blood stream infection with extended-spectrum beta lactamase-producing Enterobacter aerogenes in a neonatal intensive care unit in Fiji. The source of the outbreak was traced to a bag of contaminated normal saline in the ward, which was used for multiple patients. All isolates recovered from patients were indistinguishable from the bacteria recovered from the normal saline by pulsed-field gel electrophoresis. The outbreak was controlled using simple infection control practices such as reinforcement of strict hand hygiene policy, provision of single use vials of normal saline, and strict aseptic technique for injections. PMID- 19552519 TI - A comparison of the reactivating and therapeutic efficacy of newly developed bispyridinium oximes (K250, K251) with commonly used oximes against tabun in rats and mice. AB - The potency of newly developed bispyridinium compounds (K250, K251) in reactivating tabun-inhibited acetylcholinesterase and reducing tabun-induced lethal toxic effects was compared with currently available oximes (obidoxime, trimedoxime, the oxime HI-6) using in vivo methods. Studies determined percentage of reactivation of tabun-inhibited blood and tissue AChE in poisoned rats and showed that the reactivating efficacy of both newly developed oximes is comparable with the oxime HI-6 but it is significantly lower than the reactivating effects of obidoxime and trimedoxime, especially in diaphragm and brain. Both newly developed oximes were also found to be able to slightly reduce lethal toxic effects in tabun-poisoned mice. Their therapeutic efficacy is higher than the potency of the oxime HI-6 but it is lower than the therapeutic effects of trimedoxime and obidoxime. Thus, the reactivating and therapeutic potency of both newly developed oximes (K250, K251) does not prevail over the effectiveness of currently available oximes and, therefore, they are not suitable for their replacement for the treatment of acute tabun poisoning. PMID- 19552520 TI - A preliminary assessment of relative sensitivities to foreign red blood cell challenges in the northern bobwhite for potential evaluation of immunotoxicity. AB - Many environmental toxins have been shown to suppress the immune system across taxa. The foreign red blood cell (RBC) challenge is an important part of a complement of tests used to assess immunocompetence in the laboratory because it can assess an individual's humoral response without impacting its health. This challenge is used commonly across species and measures antibody titers in response to an intraperitoneal, intravenous, or subcutaneous injection of foreign RBCs. Determination of the best appropriate foreign RBC challenge is therefore important when designing tests for evaluation of humoral responses. The northern bobwhite (Colinus virginianus) is a commonly used species for avian toxicity tests, however little is known about the relative sensitivities of its humoral responses to foreign erythrocytes. In this pilot study, we exposed adult quail to intravenous injections of 5% solutions of sheep, rat, rabbit, bovine, or chicken erythrocytes and performed antibody titers [hemagglutination assay for total immunoglobulin (Ig), IgG, and IgM] for primary and secondary responses. Although the bobwhites appeared to respond strongly to rat RBCs, high variability in responses were observed among individuals. Chicken RBCs elicited the poorest responses for both primary and secondary challenges. Sheep and bovine RBCs were adequate antigens for this test in bobwhites. We found that rabbit erythrocytes elicited the strongest responses with the least amount of variability between individuals. Rabbit RBCs, therefore, appear to be the ideal antigen for this test of the humoral response in this species. PMID- 19552521 TI - A review of inhalability fraction models: discussion and recommendations. AB - The first step in mathematically modeling the mechanics of respiratory deposition of particles is to estimate the ability of a particle to enter the head, either through the mouth or nose. Models of the biological effects from inhaled particles are commonly, albeit incorrectly, simplified by making an assumption that the only particles of concern are those that can readily penetrate to the pulmonary region of the lung: typically particles less than 5microm in aerodynamic diameter. Inhalability for particles of this size is effectively 100%, so there is little need to develop a mathematical representation of the phenomenon. However, chemical irritants, biological agents, or radioactive material, in the form of large particles or droplets, can cause adverse biological responses by simply being taken into the head and depositing in the extrathoracic area. As a result, it is important to understand the inhalability of both small and large particles. The concept of particle inhalability received little consideration until the 1970s; since then it has been the subject of many experiments with a fairly wide disparity of results, in part due to the variety of dependent variables and the difficulty in adequate measurement methods. This article describes the currently utilized models of inhalability, recommends specific methods for implementing inhalability into mathematical models of respiratory deposition, and identifies outstanding issues and limitations. In this article, we describe inhalability as it applies to particulate matter and liquid droplets; modeling the inhalability of fibers is a work in progress and is not addressed. PMID- 19552522 TI - Autophagy: regulation and role in disease. AB - Autophagy, a lysosomal process involved in the maintenance of cellular homeostasis, is responsible for the turnover of long-lived proteins and organelles that are either damaged or functionally redundant. The process is tightly controlled by the insulin-amino acid-mammalian target of the rapamycin dependent signal-transduction pathway. Research in the last decade has indicated not only that autophagy provides cells with oxidizable substrate when nutrients become scarce but also that it can provide protection against aging and a number of pathologies such as cancer, neurodegeneration, cardiac disease, diabetes, and infections. PMID- 19552523 TI - Analysis of Plasmodium falciparum var genes expressed in children from Papua New Guinea. AB - BACKGROUND: The variable antigen P. falciparum erythrocyte membrane protein-1 (PfEMP1) is a major virulence factor in malaria. A large number of var genes encode PfEMP1, and we hypothesized that a restricted PfEMP1 repertoire determines clinical disease presentation. We conducted a case-control study in Papua New Guinea and analyzed transcribed var genes in naturally infected children. METHODS: var messenger RNA was isolated from 78 children with asymptomatic, mild, or severe malaria. We prepared complementary DNA from the upstream region into the DBL1alpha domain and picked, on average, 20 clones for sequencing. RESULTS: Twenty-five percent of centrally located var genes were shared between children, whereas only 5% of subtelomeric genes were shared, indicating lower diversity in the former group. Linkage between group B or C var upstream sequences and DBL1alpha groups was not observed, which impeded prediction by DBL1alpha analysis. A higher proportion of var group A sequences was detected in symptomatic malaria, and a subgroup of frequently encountered var genes with complex head structure seems to be associated with severe malaria. A subset of var group C genes was frequently expressed in older children with asymptomatic high levels of parasitemia. CONCLUSION: Despite this vast diversity, restricted disease-associated var genes were identified and might be used for innovative interventions based on PfEMP1. PMID- 19552524 TI - Parenteral lipid emulsion induces germination of Candida albicans and increases biofilm formation on medical catheter surfaces. AB - BACKGROUND: The administration of parenteral nutrition, including lipid emulsion (LE), to patients via medical catheters is an unexplained risk factor for the development of candidemia. Germination and biofilm formation are recognized virulence determinants of Candida albicans. No studies have addressed the effect of LE on candidal biofilm production. In this study, we investigated the effect of LE on candidal germination and its ability to form biofilm on medical catheter material. METHODS: C. albicans strain SC-5314 was grown in standard growth medium in the presence or absence a commercially available LE. Biofilms grown on silicone-elastomer catheter discs in these media were compared for mass by dry weight measurements. Biofilm morphology was analyzed by scanning electron microscopy and confocal laser microscopy. The effect of LE on C. albicans germination and growth was evaluated microscopically and by determination of colony-forming units, respectively. RESULTS: Addition of LE to standard growth medium increased C. albicans biofilm production and resulted in observed changes in biofilm morphology and architecture. Furthermore, LE induced germination and supported the growth of C. albicans. CONCLUSIONS: LE-inducible candidal virulence determinants, such as germination and enhanced biofilm production, may help to explain the increased risk of candidemia in patients receiving LE via medical catheters. PMID- 19552525 TI - Engagement of Toll-like receptor 2 on CD4(+) T cells facilitates local immune responses in patients with tuberculous pleurisy. AB - BACKGROUND: Although it has been recognized that Mycobacterium tuberculosis contains large amounts of Toll-like receptor 2 (TLR2) ligands, their direct effects on CD4(+) T cells and the clinical implications have not been determined. METHODS: With the recent finding that activated CD4(+) T cells express TLR2 as a costimulatory receptor, we hypothesized that M. tuberculosis and its components may directly affect CD4(+) T cells by engaging TLR2, thus facilitating the expansion and function of these lymphocytes in tuberculous pleura. RESULTS: Our results indicate that CD4(+) T cells from the pleural fluid and peripheral blood of patients with tuberculosis show significantly increased TLR2 expression, compared with those from healthy donors. TLR2 ligand activity was also significantly higher in the tuberculous pleural fluid than in the serum from healthy donors or patients with pulmonary tuberculosis. M. tuberculosis TLR2 ligands, 19-kDa lipoprotein, and live bacillus Calmette-Guerin all modulated cytokine production (interferon gamma and interleukin 17), cellular proliferation, survival, and migration of CD4(+) T cells isolated from pleural fluid and activated with anti-CD3 and anti-CD28. CONCLUSION: These data indicate that direct interaction between M. tuberculosis TLR2 ligands and CD4(+) T cells facilitated local CD4(+) T cell immune responses in patients with tuberculous pleurisy. PMID- 19552526 TI - Inactivation of the Haemophilus ducreyi luxS gene affects the virulence of this pathogen in human subjects. AB - Haemophilus ducreyi 35000HP contains a homologue of the luxS gene, which encodes an enzyme that synthesizes autoinducer 2 (AI-2) in other gram-negative bacteria. H. ducreyi 35000HP produced AI-2 that functioned in a Vibrio harveyi-based reporter system. A H. ducreyi luxS mutant was constructed by insertional inactivation of the luxS gene and lost the ability to produce AI-2. Provision of the H. ducreyi luxS gene in trans partially restored AI-2 production by the mutant. The luxS mutant was compared with its parent for virulence in the human challenge model of experimental chancroid. The pustule-formation rate in 5 volunteers was 93.3% (95% confidence interval, 81.7%-99.9%) at 15 parent sites and 60.0% (95% confidence interval, 48.3%-71.7%) at 15 mutant sites (1-tailed P < .001). Thus, the luxS mutant was partially attenuated for virulence. This is the first report of AI-2 production contributing to the pathogenesis of a genital ulcer disease. PMID- 19552528 TI - Disposition of GDC-0879, a B-RAF kinase inhibitor in preclinical species. AB - 1. The pharmacokinetics and disposition of GDC-0879, a small molecule B-RAF kinase inhibitor, was characterized in mouse, rat, dog, and monkey. 2. In mouse and monkey, clearance (CL) of GDC-0879 was moderate (18.7-24.3 and 14.5 +/- 2.1 ml min(-1) kg(-1), respectively), low in dog (5.84 +/- 1.06 ml min(-1) kg(-1)) and high in rat (86.9 +/- 14.2 ml min(-1) kg(-1)). The volume of distribution across species ranged from 0.49 to 1.9 l kg(-1). Mean terminal half-life values ranged from 0.28 h in rats to 2.97 h in dogs. Absolute oral bioavailability ranged from 18% in dog to 65% in mouse. 3. Plasma protein binding of GDC-0879 in mouse, rat, dog, monkey, and humans ranged from 68.8% to 81.9%. 4. In dog, the major ketone metabolite (G-030748) of GDC-0879 appeared to be formation rate limited. 5. Based on assessment in dogs, the absorption of GDC-0879 appeared to be sensitive to changes in gut pH, food and salt form (solubililty), with approximately three- to four-fold change in areas under the curve (AUCs) observed. PMID- 19552529 TI - Species differences for stereoselective metabolism of ethofumesate and its enantiomers in vitro. AB - 1. The stereoselective metabolism of ethofumesate (ETO) and its enantiomers in rabbit and rat liver microsomes have been studied by chiral high-performance liquid chromatography (HPLC) method. Two metabolites were detected in both liver microsomes in the presence of beta-nicotinamide adenine dinucleotide phosphate (NADPH). 2. The T(1/2) of (+)-ETO and (-)-ETO in rabbit liver microsomes were 12.2 and 4.7 min of rac-ETO and 25.9 and 6.7 of ETO enantiomers. However, the T(1/2) of (+)-ETO and (-)-ETO in rat liver microsomes were 5.3 and 5.9 min of rac ETO and 7.8 and 10.6 of ETO enantiomers. The stereoselective selectivity is similar to the in vivo study. 3. After incubation of ETO enantiomers, stereoselectivity was present in the formation of ETO-OH enantiomer in rabbit liver microsomes, but stereoselectivity was not evident in rat liver microsomes. 4. There was no chiral inversion from the (+)-ETO to (-)-ETO or inversion from ( )-ETO to (+)-ETO in both rabbit and rat liver microsomes. PMID- 19552530 TI - Particles from wood smoke and road traffic differently affect the innate immune system of the lung. AB - The effect of particles from road traffic and wood smoke on the innate immune response in the lung was studied in a lung challenge model with the intracellular bacterium Listeria monocytogenes. Female Balb/cA mice were instilled intratracheally with wood smoke particles, particles from road traffic collected during winter (studded tires used; St+), and during autumn (no studded tires; St ), or diesel exhaust particles (DEP). Simultaneously with, and 1 or 7 days after particle instillation, 10(5) bacteria were inoculated intratracheally. Bacterial numbers in the lungs and spleen 1 day after Listeria challenge were determined, as an indicator of cellular activation. In separate experiments, bronchoalveolar lavage (BAL) fluid was collected 4 h and 24 h after particle instillation. All particles tested reduced the numbers of bacteria in the lung 24 h after bacterial inoculation. When particles were given simultaneously with Listeria, the reduction was greatest for DEP, followed by St+ and St-, and least for wood smoke particles. Particle effects were no longer apparent after 7 days. Neutrophil numbers in BAL fluid were increased for all particle exposed groups. St+ and St- induced the highest levels of IL-1beta, MIP-2, MCP-1, and TNF-alpha, followed by DEP, which induced no TNF-alpha. In contrast, wood smoke particles only increased lactate dehydrogenase (LDH) activity, indicating a cytotoxic effect of these particles. In conclusion, all particles tested activated the innate immune system as determined with Listeria. However, differences in kinetics of anti-Listeria activity and levels of proinflammatory mediators point to cellular activation by different mechanisms. PMID- 19552531 TI - Decreased urinary secretion of belotecan in folic acid-induced acute renal failure rats due to down-regulation of Oat1 and Bcrp. AB - The effects of folic acid-induced acute renal failure on the renal excretion of belotecan were investigated in rats after intravenous administration. Both glomeruli and renal tubules were seriously damaged by folic acid-induced acute renal failure. The renal excretion clearance, CLr, of belotecan was significantly decreased by folic acid-induced acute renal failure. Furthermore, glomerular filtration rate and secretion clearance of the drug were dramatically decreased by folic acid-induced acute renal failure. In vivo renal uptake of belotecan was inhibited by p-aminohippurate, whereas renal excretion was inhibited by GF120918, but not by verapamil and bromosulphalein. This indicates that Oat1/3 and Bcrp are involved in the renal uptake and urinary excretion of belotecan, respectively. Both mRNA and protein levels of Oat1, Oat3 and Bcrp were significantly decreased in folic acid-induced acute renal failure rats. Based on the finding that belotecan is a substrate of OAT1 but not of OAT3, the decrease in CLr of belotecan in folic acid-induced acute renal failure could, therefore, mainly be attributed to the down-regulation of Oat1 and Bcrp, in addition to the decrease in glomerular filtration rate. PMID- 19552540 TI - A paired comparison between human skin and hairless guinea pig skin in vitro permeability and lag time measurements for 6 industrial chemicals. AB - The purpose of the present study was to measure and compare permeability coefficients (k(p)) and lag times (tau) in human skin and hairless guinea pig (HGP) skin. Paired experiments employed heat-separated epidermal membranes from human and HGP sources mounted on static in vitro diffusion cells. Infinite-dose, saturated aqueous solutions of 6 industrial chemicals were used as donors: aniline, benzene, 1,2- dichloroethane, diethyl phthalate, naphthalene, and tetrachloroethylene. No significant differences were found between human and HGP skin for either k(p) or tau for any of these chemicals (p >or= .24). HGP vs. human k(p) measurements, and HGP vs. human tau measurements, were highly correlated. For k(p), the slope of the linear correlation was close to unity (1.080 +/- 0.182) and the intercept close to 0 (0.015 +/- 0. 029 cm/h), with a correlation coefficient (r(2)) = 0.898. For tau, the slope was also close to unity (0.818 +/- 0.030) and the intercept close to 0 (-0.014 +/- 0.023 h), with r(2) = 0.994. These results suggest that HGP skin may serve as an excellent surrogate for human skin in in vitro dermal penetration studies. PMID- 19552541 TI - Cross-reactions among hair dye allergens. AB - BACKGROUND: p-Phenylenediamine (PPD) is an important hair dye allergen, but there remains a reasonable suspicion that other hair dye chemicals may also be responsible for a proportion of the clinical burden of hair dye allergy. OBJECTIVE: To assess to what extent presently assessed additional patch test agents contribute to the diagnosis of non-PPD hair dye allergy. METHODS: A retrospective analysis was conducted of patch test results with hair dye allergens, focusing on the extent to which patients who were positive for allergic reactions to other hair dye allergens also had a concomitant positive reaction to PPD. RESULTS: For the hair dye allergens other than p-toluenediamine (PTD), reactions in the absence of a concomitant positive reaction to PPD were very rare. Positive reactors to PTD were also positive for reactions to PPD in 5 of every 6 cases. Pyrogallol positives often occurred in the absence of a PPD positive, but were never judged to be of clinical relevance. CONCLUSIONS: Hair dye chemicals other than PPD may be of importance, but the presently tested materials, with the possible exception of PTD, are normally positive only when a PPD-positive reaction is also present, suggesting that their use in patch testing in hair dye allergy is likely to be of limited value. PMID- 19552542 TI - Production and optimization of valproic acid nanostructured lipid carriers by the Taguchi design. AB - The objective of this study was production of nanostructured lipid carriers (NLC) of valproic acid for brain delivery thought the nasal pathway. NLC were prepared by solvent diffusion method using a Taguchi design. The lipid, oil and valproic acid were dissolved in organic phase then dispersed in aqueous phase containing the surfactant. The most effective factors on size were surfactant concentration, organic/aqueous phase ratio and acetone/ethanol volume ratio. Zeta potential was more affected by the lipid type, while the surfactant concentration and sonication time were more effective variables on drug release rate. The entrapment efficiency was more affected by lipid/drug weight ratio. Optimum formulation obtained with 400 mg lipid, 400 mg valproic acid, 2% surfactant, 1:5 organic/aqueous phase ratio, 1:1 acetone/ethanol volume ratio and 2 min sonication. PMID- 19552544 TI - Formulation and evaluation of a pulsatile drug delivery system using time- and pH dependant polymers. AB - The aim of the present study was to develop fast-release enteric-coated tablets for pulsatile drug delivery to the colon. The novelty of this work is a combination of pH- and time-dependant enteric polymers as a single coating. Eudragit S100 was used as a pH-dependant polymer and eudragit RL100 was used as a time-dependant polymer. Theophylline was taken as a model drug. Dissolution studies of enteric-coated tablets were performed with different media having a pH of 1.2, 7.4, and 6.8. Results of the dissolution data show that drug release in the colon could be controlled by using eudragit RL100 eudragit S100. The lag time prior to the drug release was highly affected by a combination of two factors: The percentage of eudragit RL100 and coating level. The optimum formulation was found to be one containing eudragit RL100 and eudragit S100 with a ratio of 60:40 of polymer and coating level of 4.66% w/w. The present study demonstrates that the theophylline enteric-coated tablets could be successfully formulated as a pulsatile drug delivery by the design of a time- and pH-dependant modified chronopharmaceutical formulation. In conclusion, pulsatile drug release over a period of 2-12 hours, consistent with the requirements for chronopharmaceutical drug delivery, can be achieved by using time- and pH-dependant polymers. PMID- 19552545 TI - Development and evaluation of ocular drug delivery system. AB - The eye presents unique opportunities and challenges when it comes to the delivery of pharmaceuticals. In the present study, ocular inserts of levofloxacin were prepared using chitosan and gelatin by solvent casting technique with an aim to improve therapeutic efficacy in the treatment of conjunctivitis. Prepared ocular inserts were then evaluated for film thickness, weight variation, content uniformity, percentage moisture loss and absorption. In vitro drug release studies were carried out using flow through apparatus that simulated the eye conditions. Optimized formulations were subjected to in vivo and stability studies to assess the effectiveness of the formulations. Finally in vitro in vivo correlation was established. Plasticizer like PEG was found to influence their effect on drug release. Prepared ocular inserts exhibited zero order kinetics which was confirmed by strong and positive correlation. The in vitro and in vivo drug release studies revealed that the formulations provide a best alternative to prolong the drug release at the end of 24 h and remained stable with intact at ambient conditions. PMID- 19552546 TI - Nanoemulsion: a promising tool for solubility and dissolution enhancement of celecoxib. AB - The solubility and dissolution of the poorly soluble drug celecoxib (CXB) was enhanced using many techniques like nanoemulsion, solid lipid nanoparticle and solid dispersion in the present investigation. The solubility of CXB in each formulation was determined using the reported HPLC method at the wavelength of 250 nm. Dissolution studies of pure CXB and its formulations were performed using USP dissolution apparatus in distilled water. The highest solubility (228. 24 mg/mL) as well as % dissolution (99.9) of CXB was obtained with nanoemulsion technique. The results of solubility and dissolution were highly significant using the nanoemulsion technique as compared to other techniques (P < 0.01). All three formulations showed a sustained type of drug release. The best sustained type drug release was obtained with nanoemulsion. This indicated that nanoemulsion can be successfully used for sustained and controlled drug delivery of CXB. Overall these findings suggested that nanoemulsion is a promising vehicle for solubility and dissolution enhancement of CXB. PMID- 19552551 TI - Physicochemical investigation of the solid dispersion systems of etoricoxib with poloxamer 188. AB - Solid dispersion systems of a poorly water-soluble drug, etoricoxib were prepared with poloxamer 188 in 1:0.5, 1:1.5 and 1:2.5 ratios and evaluated by FTIR, powder XRD and dissolution studies. Physical studies demonstrated a strong hydrogen bonding with significant decrease in the crystallinity and formation of amorphous etoricoxib in its binary systems. All binary systems of etoricoxib showed faster dissolution than pure drug alone (P < 0.001). However, 1:2.5 proportion of etoricoxib: poloxamer 188 showed superior performance (DE45: 71.27% +/- 3.85) in enhancing solubility and dissolution rate of etoricoxib suggesting optimum ratio of the carrier. PMID- 19552527 TI - Predictive value of HIV-1 genotypic resistance test interpretation algorithms. AB - BACKGROUND: Interpreting human immunodeficiency virus type 1 (HIV-1) genotypic drug-resistance test results is challenging for clinicians treating HIV-1 infected patients. Multiple drug-resistance interpretation algorithms have been developed, but their predictive value has rarely been evaluated using contemporary clinical data sets. METHODS: We examined the predictive value of 4 algorithms at predicting virologic response (VR) during 734 treatment-change episodes (TCEs). VR was defined as attaining plasma HIV-1 RNA levels below the limit of quantification. Drug-specific genotypic susceptibility scores (GSSs) were calculated by applying each algorithm to the baseline genotype. Weighted GSSs were calculated by multiplying drug-specific GSSs by antiretroviral (ARV) potency factors. Regimen-specific GSSs (rGSSs) were calculated by adding unweighted or weighted drug-specific GSSs for each salvage therapy ARV. The predictive value of rGSSs were estimated by use of multivariate logistic regression. RESULTS: Of 734 TCEs, 475 (65%) were associated with VR. The rGSSs for the 4 algorithms were the variables most strongly predictive of VR. The adjusted rGSS odds ratios ranged from 1.6 to 2.2 (P < .001). Using 10-fold cross validation, the averaged area under the receiver operating characteristic curve for all algorithms increased from 0.76 with unweighted rGSSs to 0.80 with weighted rGSSs. CONCLUSIONS: Unweighted and weighted rGSSs of 4 genotypic resistance algorithms were the strongest independent predictors of VR. Optimizing ARV weighting may further improve VR predictions. PMID- 19552552 TI - Properties of theophylline tablets dry powder coated with Eudragit((R)) E PO and Eudragit((R)) L 100-55. AB - The aim of the present study was to investigate the influence of Eudragit((R)) E PO on the drug release mechanism of Eudragit((R)) L 100-55 film coatings applied to theophylline tablets by a dry powder coating technique. The process was entirely liquid-free. Calculation of the Flory-Huggins interaction parameter based on solubility parameters suggested immiscibility of the two copolymers. MDSC thermograms were characterized by two glass transitions for the investigated Eudragit((R)) E PO/Eudragit((R)) L 100-55 ratios and confirmed incomplete miscibility of the copolymers at processing conditions. FT-IR analysis was employed to study binding interactions of the polymers. Due to the higher affinity of the plasticizer, triethyl citrate, for Eudragit((R)) E PO compared to Eudragit((R)) L 100-55, redistribution of the plasticizer was observed during the curing phase of the process. Plasticizer migration also affected the initial phase of drug release from powder-coated theophylline tablets that were stored for four weeks. Drug release from powder-coated tablets was dependent on the polymer blend ratio, coating thickness, and the pH of the dissolution medium. A broad range of pH dependent theophylline release profiles were obtained as a function of the polymer blend ratio. The particle size of the coating powder influenced the microstructure of the film coating. PMID- 19552554 TI - Formulation of intelligent tablets with an antacid effect. AB - Matrix systems with a local antacid effect were produced in this study. Aluminium hydroxide and magnesium trisilicate in constant concentrations were used as active agents. Eudragit E PO was applied as a matrix former and sodium bicarbonate as a disintegrant (third antacid component), in different ratios. Their effects on the properties of the tablets were studied. Such formulated systems must be insoluble if the pH of the stomach is less acidic, but a rapid disintegration must occur if necessary. It can be concluded that Eudragit E PO in appropriate composition can ensure tablets with pH-dependent disintegration. Its binding effect allows tablet making from the elastic active component. The liberation of antacid materials from this system is controlled. If the pH reached 2.5, the erosion of the tablet was reduced. In contrast with expectations, the application of poorly compressible and effervescent sodium bicarbonate increased the time for disintegration of the tablets, because of its extended alkalizing effect around the tablet. This system with this acrylic component is appropriate to produce a controlled-release local antacid preparation. PMID- 19552558 TI - Influence of formulation composition and processing on the content uniformity of low-dose tablets manufactured at kilogram scale. AB - The purpose of this study was to investigate the impact of processing, API loading, and formulation composition on the content uniformity of low-dose tablets made using direct compression (DC) and roller compaction (RC) methods at 1 kg scale. Blends of 1:1 microcrystalline cellulose/lactose or 1:1 microcrystalline cellulose/dicalcium phosphate anhydrous with active pharmaceutical ingredient (API) at loadings of 0.2, 1 and 5% were processed either by DC or RC. A statistical analysis showed that DC produced comparable content uniformity results to RC. Microcrystalline cellulose/lactose formulations had improved average potency compared to microcrystalline cellulose/dicalcium phosphate anhydrous formulations for both DC and RC. The impact of segregation in the DC blends and adhesion to equipment surfaces was assessed to aid in understanding potency trends. DC may be as suitable as RC for low-dose regime (e.g. < 1 mg) when manufacturing clinical supplies at small scale provided the API has a suitable particle size and potency loss to equipment is negligible. PMID- 19552560 TI - Use of scoring to induce reproducible drug delivery from osmotic pulsatile tablets. AB - An osmotic-controlled pulsatile delivery technology was developed for targeted drug delivery. This novel system consists of a tablet core surrounded by an osmotic coating that has been mechanically compromised in strategic locations to facilitate reliable drug release at a given time point after administration. The tablet core contains a high drug load in addition to several osmotic agents and swellable polymers, and the surrounding mechanically-compromised osmotic coating consists of a semipermeable membrane that has been scored with a razor blade in several key locations. The components in the tablet core attract water into the core, causing it to swell and propagate the scores in the coating along the length of the tablet. After the scores have fully propagated, the coating bursts open, releasing the tablet core's contents, including the drug, into the surrounding media. The variables that were investigated in this study included the configuration of the scores in the coating, the length of the scores, and the distance between the scores. The delivery system developed in this work is able to generate a reproducible dissolution profile consisting of a specific targeted lag time, between five minutes and two hours, followed by immediate release of the drug from the core. The performance of the system was validated in vitro using the drug salicylic acid. Unlike previously developed osmotic pulsatile delivery systems, the present system is able to accommodate higher drug loading levels, it is easier to manufacture, and has demonstrated more reproducible burst times (i.e. burst time) than several other pulsatile systems. PMID- 19552561 TI - Bioequivalence studies and sugar-based excipients effects on the properties of new generic ketoconazole tablets formulations and stability evaluation by using direct compression method. AB - In this work we described the development of a new solid oral formulation of ketoconazole, a broad-spectrum antifungal agent that belongs to the class II of Biopharmaceutics Classification System (BCS). The ketoconazole raw material supplier was selected to present a best flow and compactation. In addition we used direct compression and superdisintegrants associated to polyols to enhance the dissolution of the ketoconazole tablets. The dissolution was evaluated based in level C in vivo/in vitro correlation established. The best formulation was obtained with croscarmellose/maltose association that in the accelerated stability assays presented no differences on quality specifications and no drug excipients interaction by DSC analyses. In this work it was possible to confirm the use of sugar-based excipients as suitable dissolution enhancers in pharmaceutical technology and real processes conditions. PMID- 19552562 TI - Egg shell membrane as a substrate for optimizing in vitro transbuccal delivery of glipizide. AB - Buccoadhesive gels for transbuccal delivery of glipizide were prepared using different bio-adhesive polymers. The gels were prepared by solution polymerization technique. An apparatus simulating the in vivo conditions of the mouth was designed in order to assess in vitro drug release kinetics of these gels. The gels were also evaluated for spreadability, buccoadhesive strength, swelling index, and viscosity. Maximum buccoadhesive strength was observed for formulation, F8 with good sustained release behavior, whereas viscosity and swelling index was highest for the formulation, F5 but with minimum buccoadhesive strength. The drug release kinetics followed Higuchi model with release mechanism being Fickian diffusion. PMID- 19552564 TI - Enhancement of transdermal delivery of progesterone using medium-chain mono and diglycerides as skin penetration enhancers. AB - We evaluated whether medium-chain mono and diglycerides (MCG) can be utilized to optimize the transdermal delivery of progesterone (PGT). MCG was studied at 10 70% (w/w) in propylene glycol (a polar solvent) or Myvacet oil (nonpolar solvent); PGT was used at 1% (w/w). The topical (to the skin) and transdermal (across the skin) delivery of PGT were evaluated in vitro using porcine ear skin. When incorporated in propylene glycol, MCG at 10% enhanced the topical and transdermal delivery of PGT by 2.5- and 7-fold, respectively. At 20-50%, topical delivery was further enhanced while transdermal delivery gradually returned towards baseline. At 70%, MCG enhanced neither the delivery to viable skin nor the transdermal delivery of PGT. Similar concentration-dependent effects were observed when MCG was incorporated in Myvacet oil, but their magnitudes were 2- to 3-fold smaller. The relative safety of MCG was assessed in cultured fibroblasts and compared to propylene glycol (regarded as safe) and sodium lauryl sulfate (moderate-to-severe irritant). Both MCG and propylene glycol were substantially less cytotoxic than sodium lauryl sulfate. We conclude that formulations containing 10% MCG in propylene glycol may be a simple and safe method to improve the transdermal delivery of progesterone and promote its use in hormone replacement therapy. PMID- 19552563 TI - Formulation and process development of (recombinant human) deoxyribonuclease I as a powder for inhalation. AB - A formulation and process development study was performed to formulate recombinant human deoxyribonuclease I as a powder for inhalation. First, excipient compatibility (with bovine DNase as a model substance) was examined with a stability study at stressed conditions (60 and 85 degrees C) while monitoring for occurrence of the Maillard reaction. Next, powders for inhalation were prepared by spray drying and spray freeze drying. We found that spray drying with inulin as stabilizer resulted in the best powder for inhalation. Finally, an ex-vivo test with the spray dried rhDNase I/inulin powder significantly decreased elastic and viscous moduli of sputum from five cystic fibrosis patients. PMID- 19552565 TI - Evaluation of Blatta orientalis (Q) nasal gel formulation in milk aspiration induced eosinophilia. AB - BACKGROUND: The purpose of the present study was to develop intranasal delivery systems of the homeopathic anti-asthmatic remedy Blatta orientalis mother tincture (Q) using thermoreversible polymer Pluronic F127 (PF127) and mucoadhesive polymer Carbopol 934P (C934P). METHODS: Formulations were modulated so as to have a gelation temperature below 34 degrees C to ensure gelation at physiological temperature after intranasal administration. Its gelation temperature, mucoadhesive strength, viscosity and gel strength were studied. B. orientalis (Q) nasal gel was tested with recurrent milk aspiration to determine whether it produces changes in eosinophilia in a murine model of asthma. RESULT: The gelation temperatures of the formulations and mucoadhesive strength, determined using sheep nasal mucosal membrane, increased by the addition of increasing concentrations of Carbopol. The results of milk aspiration induced eosinophilia, B. orientalis (Q) nasal gel significantly (P < 0.001), decreased eosinophil cell count as compared with toxicant by using in absolute eosinophilia count method. Finally, histopathological examination did not detect any damage during in vivo studies. CONCLUSION: The PF127 gel formulation of B. orientalis (Q) with in situ gelling and mucoadhesive properties with increased permeation rate is promising for prolonging nasal residence time and thereby nasal absorption. PMID- 19552566 TI - Lack of association of angiotensin-converting enzyme insertion/deletion polymorphism and myocardial infarction at very young ages. AB - We examined whether angiotensin-converting enzyme (ACE) insertion (I)/deletion (D) polymorphism is associated with the development of myocardial infarction (MI) at < or = 35 years of age. The study sample consisted of 201 patients with premature MI and 140 age- and sex-matched healthy individuals. No difference was found in the distribution of ACE genotypes between the patients and controls. A higher prevalence of the DD genotype among hypertensives was found compared with the non-hypertensive patients (62.5% vs 35.6%, p = 0.01). ACE polymorphism is not associated with the development of premature MI and this might be due to the low prevalence of hypertension in young coronary patients. PMID- 19552567 TI - Serum markers in pre-eclampsia. AB - Preeclampsia occurs approximately in 10% of pregnancies and remains a leading cause of maternal and neonatal mortality and morbidity worldwide, particularly in developing countries. The condition is usually diagnosed in late pregnancy by the presence of hypertension with proteinuria and/ or edema. Prevention of any disease process requires knowledge of its etiology and pathogenesis, as well as the availability of methods for prediction of those at high risk for this disorder. Numerous clinical, biophysical, and biochemical tests have been proposed for prediction or early detection of preeclampsia. This review will explore the current tests available in the evaluation of hypertensive complications of pregnancy. PMID- 19552568 TI - Development and set-up of a portable device to monitor airway exhalation and deposition of particulate matter. AB - The aim of this study was to assess and monitor airway exhalation and deposition of particulate matter (PM). After standardizing inspiratory/expiratory flow and volumes, a novel device was tested on a group of 20 volunteers and in a field study on workers exposed to cristobalite. Both male and female subjects showed a higher percentage of deposition in the 0.5 microm channel than in the 0.3 microm channel on a laser particle counter, but it was higher in the males because of their higher exhaled lung volumes. The device was tested on a wider range of particles (0.3-0.5-1.0-2.5 microm) in the cristobalite productive division. The device has low intrasubject variability and good reproducibility, with geometric mean of %CV < 5%. Such a measure can be used to assess individual susceptibility to PM, making repeated measures in different environments, and examining the persistence of particles in the airways after a period in polluted environments. PMID- 19552569 TI - New potential biomarkers in the diagnosis of esophageal squamous cell carcinoma. AB - OBJECTIVE: To analyse the alterations of serum proteins in cases of esophageal squamous cell carcinoma (ESCC) in order to screen and validate serum marker patterns for the diagnosis of ESCC in the high-risk populations of Xinjiang, China. METHODS: The serum proteomic patterns of 188 cases, including 139 patients with ESCC (54 Uygur, 45 Kazakh and 40 Han subjects) and 49 sex- and age-matched healthy controls, were detected using the SELDI-TOF-MS (surface-enhanced laser desorption/ionization-time of flight-mass spectrometry) technology with the CM10 ProteinChip. Differences in protein peaks between patients with ESCC and controls were analysed using the Biomarker Pattern Software, and a primary diagnosis model of ESCC was developed and validated with SVM (support vector machines). This model was further evaluated by a large-scale blind test. RESULTS: Two hundred and eighty-three protein peaks were detected within the molecular range of 0-20 kDa, among which, 140 peaks were significantly different between ESCC cases and controls (p < 0.05). A diagnostic pattern consisting of six protein peaks (m/z 5667, 5709, 5876, 5979, 6043 and 6102) was established with a sensitivity of 97.12% and a specificity of 83.87%. The large-scale blind test generated a sensitivity of 91.43% and a specificity of 88.89%. CONCLUSIONS: The differential protein peaks analysed by SELDI-TOF-MS may contain promising serum biomarkers for screening ESCC. The diagnostic model which combined only six protein peaks had a satisfactory discriminatory power. The model should be further evaluated in other populations of ESCC patients and tested against controls. The nature and function of the discriminating proteins have yet to be elucidated. PMID- 19552571 TI - Increases in serum concentration of human heart-type fatty acid-binding protein following elective coronary intervention. AB - BACKGROUND: Heart-type fatty acid-binding protein (H-FABP) is considered a marker of myocardial necrosis but whether or not it is modified by myocardial ischemia is not clear. We sought to investigate if H-FABP serum levels increase following non-urgent coronary angioplasty. METHODS: We studied 31 patients undergoing coronary angioplasty. Peripheral venous samples were drawn immediately before angioplasty, 1 h after the first balloon inflation and 24 h after the procedure and assayed for H-FABP. RESULTS: Serum levels of H-FABP increased significantly at 1 h vs baseline from 2554 +/- 1268 to 3322 +/- 245 pg ml(-1) (p = 0.024). However, no differences were observed between 1 h and 24 h after angioplasty (3268 +/- 1861 vs 3322 +/- 2459 pg ml(-1), p = 0.87). Moreover, no significant difference was observed when we compared 24 h after angioplasty with the baseline (3268 +/- 1861 vs 2554 +/- 1268 pg ml(-1), p = 0.112). CONCLUSIONS: We conclude that H-FABP significantly increases after elective coronary angioplasty at 1 h compared with baseline values; whether or not this has any prognostic significance for future events, as it occurs with troponins, needs to be studied further. PMID- 19552570 TI - Thrombin activity throughout the acute phase of acute ST-elevation myocardial infarction and the relation to outcome. AB - BACKGROUND: Thrombin and plasmin play a central role in ongoing thrombosis and platelet activation in patients with acute ST-elevation myocardial infarction (STEMI). Data of thrombin and plasmin activity in the early course of STEMI and the relation to outcome are scarce. METHODS: We included 68 consecutive patients (53 male, 59 +/- 11.4 years) with STEMI who underwent acute catheter-based reperfusion therapy within the first 12 h after onset of symptoms. Blood samples were taken at admission and after 4, 8, 12 and 24 h. Thrombin activity and generation was measured by changes in the thrombin/antithrombin-III complex (TAT) and prothrombin fragment (F1.2); plasmin was measured by changes in the plasmin alpha(2)/antiplasmin complex (PAP). A follow-up with respect to the combined primary endpoint consisting of death, acute myocardial infarction or urgent need for revascularization up to 6 weeks post-discharge was carried out. RESULTS: TAT values showed no significant change over time in patients with and without the primary endpoint but there was a borderline difference between these groups at 4 h after admission (event group 9.0 vs no event group 4.7 microg l(-1), p = 0.057). F1.2 values were different between groups only after 24 h (event group 1.5 vs no event group 0.9 nmol l(-1), p = 0.028) and did not differ in serial sampling of 24 h. PAP values were higher in patients with events after 4 and 8 h and declined over time in the group without events (p <0.001). Odds ratios (OR) with respect to the primary endpoint were highest for TAT >4.8 microg l(-1) at 0 h and TAT >8.4 microg l(-1) at 4 h (OR 7.1, 95% confidence interval (CI) 1.5-34, p = 0.015 and OR 5.5, 95% CI 1.5-20.0, p = 0.01, respectively). The predictive value of plasmin concentrations were equally high after 4 h (PAP >962 microg l( 1); OR 6.8, 95% CI 1.8-26.2, p = 0.005) and 8 h (PAP >495 microg l(-1), OR 6.7, 95% CI 1.4-32.9, p = 0.024). Values for F1.2 were only predictive after 24 h (F1.2 >0.85 nmol l(-1), OR 13, 95% CI 1.4-117.8, p = 0.023). CONCLUSIONS: Markers of thrombin and plasmin activity in acute STEMI are related to outcome. The marker for thrombin generation F1.2 becomes a significant predictor of outcome at 24 h after admission, reflecting the potentially adverse effects of ongoing thrombin generation. This underlines the potential for direct thrombin inhibition and individualization of treatment by thrombin markers in STEMI. PMID- 19552578 TI - Modification of wool surface by liposomes for dyeing with weld. AB - In this research work, wool surface has been modified by liposome to investigate its effects on dyeing with weld, a yellow natural dye. To do this, samples were first treated with aluminium sulphate and afterward with different concentrations of liposomes at various temperatures for 30 minutes and, finally, dyed with weld at 75, 85, and 95 degrees C for 30, 45, and 60 minutes. K/S values of fabric samples were calculated and washing, light and rub fastness properties of the samples were indicated. The results proposed that the sample treated with 1% liposomes and dyed at 75 degrees C for 60 min has the highest K/S value. The central composite design (CCD) used for the experimental plan with three variables on the results of color strength and statistical analysis confirms the optimum conditions obtained by the experimental results. It was also found that washing, light, wet, and dry rub fastness properties of samples dyed with weld, including liposomes, have not significantly changed. The results of water drop absorption indicated that the hydrophobicity is higher for the samples pretreated with liposomes. The SEM picture of wool sample treated with mordant and liposomes and finally dyed with weld shows a coated layer on the fiber surface. PMID- 19552579 TI - Solid lipid nanoparticles and nanoemulsions containing ceramides: preparation and physicochemical characterization. AB - Nanoemulsions (NEs) and solid lipid nanoparticles (SLNs) are widely used colloidal carriers for bioactive compounds. They are used in therapeutic, diagnostic, and cosmetic formulations. Ceramides are main components of the stratum corneum and are essential for the efficient barrier function. Their very high lipophilicity renders them difficult to incorporate in an acceptable formulation. The aim of this work was to investigate the possibility of using the benefits of nanotechnology in the efficient topical delivery of ceramides formulated as NEs or SLNs. The physicochemical characteristics of such carriers incorporating ceramides were investigated and their stability over time was assessed. Their morphology was examined under a scanning electron microscope and the interactions of their components were studied by differential scanning calorimetry. The results showed that the nanoemulsions can incorporate a high percentage (48.4% of total lipids by weight) of ceramides giving more homogeneous particle distributions of spherical-shaped nanoparticles and they maintained their characteristics over time. On the contrary, SLNs' incorporation of ceramide higher than 10.8% of total lipids by weight led to the formation of rod-like nanoparticles deteriorating the homogeneity of the particle distribution, as depicted on the high polydispersity indexes of the corresponding formulations. The results demonstrate that NEs may be the more suitable carrier, compared to SLNs. PMID- 19552583 TI - Asian sand dust aggravates allergic rhinitis in guinea pigs induced by Japanese cedar pollen. AB - Asian sand dust (ASD) contains microbial materials, sulfate (SO(4)(2-)), and nitrate (NO(3)(-)), and is derived from air pollutants in East China. ASD reportedly causes adverse respiratory health effects; a case in point is aggravated allergen-associated experimental lung eosinophilia. Guinea pigs were administered normal saline (control), ASD (0.3 mg/animal), ASD (0.6 mg/animal), Japanese cedar pollen (JCP) (0.2 mg/kg body weight), JCP + ASD (0.3 mg/animal), or JCP + ASD (0.6 mg/animal), into their nasal cavities at seven weekly intervals. The number of sneezes, amount of nasal secretions, and nasal obstructing response were measured as indices of nasal responses. Total immunoglobulin E (IgE) antibodies in serum and the number of eosinophils, histamine, and arachidonic acid metabolites in nasal cavity lavage fluids (NCLF) were also measured. ASD enhanced the JCP-associated nasal obstructing response, but not the number of sneezes or amount of nasal secretions. ASD enhanced JCP associated cysteinyl leukotrienes (C(4), D(4), E(4)) and histamine production in NCLF. ASD augmented the number of eosinophils in NCLF and total IgE in serum induced by JCP. ASD enhanced eosinophil recruitment in the nasal mucosa, and goblet cell proliferation in the nasal epithelium induced by JCP. These results suggest that ASD enhances the nasal allergic reaction induced by repeated JCP administration in guinea pigs. PMID- 19552588 TI - Re: Use of a warming bath to prevent lens fogging during laparoscopy. (From Brown JA, Inocencio MD, and Sundaram CP. J Endourol 2008;22:2413). PMID- 19552585 TI - Dynamic oxygen enhances oocyte maturation in long-term follicle culture. AB - Traditionally, follicles have been grown in standard incubators with atmospheric oxygen concentration. However, preantral follicles exist in the avascular cortex of the ovary. This study examines the effectiveness of an oxygen delivery protocol that more closely mimics the in vivo environment of the ovary on oocyte viability, maturation, parthenogenetic activation, and fertilization from in vitro cultured rat preantral follicles. Of 54 oocytes cultured in the dynamic oxygen environment, 35 were viable while only 22 of 50 oocytes cultured within an ambient oxygen concentration remained viable (p < 0.05). Germinal vesicle breakdown was observed in 56% of oocytes from the dynamic oxygen group compared to 30% of oocytes from the ambient oxygen group (p < 0.05). Parthenogenetic activation was observed in a significant number of oocytes from the dynamic oxygen group, while none of the oocytes from the ambient oxygen group activated (p < 0.05). However, the proportions of oocytes from the dynamic oxygen group that remained viable underwent germinal vesicle breakdown, and activated were still significantly less than those from the in vivo control group (p < 0.05). Fertilization of the oocytes from the dynamic oxygen group was confirmed through a successful trial of intracytoplasmic sperm injection. PMID- 19552590 TI - Cost-effectiveness analysis of raltegravir in treatment-experienced HIV type 1 infected patients in Spain. AB - Raltegravir, a novel HIV-1 integrase inhibitor, has superior efficacy with optimized background treatment (OBT) vs. placebo + OBT in treatment-experienced HIV-1 patients. This study assessed the long-term cost effectiveness of raltegravir from a Spanish National Healthcare System perspective. A cohort-state transition model was used to estimate clinical and economic outcomes associated with raltegravir + OBT vs. OBT alone. Subjects were stratified into health states according to HIV RNA level, CD4 count, and opportunistic infection (OI) history, and could transition into different health states over time based on projected long-term efficacy. Each health state was associated with a distinct treatment cost and utility (QoL) score. Model inputs for mortality, resource utilization, unit costs, OI risk, and long-term durability of viral suppression were obtained from clinical trials, published studies, and database analyses. Model outcomes were reported as incremental cost-effectiveness ratios (ICERs) in 2007 Euros per quality-adjusted life-year (euro/QALY) gained. Costs and QALYs were discounted at 6% per year based on Spanish cost-effectiveness guidelines. Extensive sensitivity analyses were conducted. Five years of treatment with raltegravir + OBT resulted in an additional 4.5 years of undiscounted life expectancy vs. OBT alone. The ICER of raltegravir + OBT vs. OBT alone was euro22,908/QALY and euro31,431/QALY for 3- and 5-year use, respectively. Lower ICERs were observed with lower discount rates (3%) for costs and benefits, lower raltegravir price (20%), and shorter treatment duration (3 years). ICER was also sensitive to analytical time horizon and alternative sources of QoL scores. In treatment-experienced Spanish patients, raltegravir was projected to provide survival benefits and be cost effective. PMID- 19552591 TI - Characterization of genetically diverse HIV type 1 from a London cohort: near full-length genomic analysis of a subtype H strain. AB - HIV-1 is characterized by an exceptional level of sequence diversity and a rapid rate of evolution. HIV diversity has implications for reliability of assays designed to detect and monitor infection, pathogenesis, disease progression, response to antiviral therapeutics, resistance pathways, and vaccine development. In the present study, HIV-1 strain diversity was assessed for a small clinical cohort (n = 15) from London, England at risk for infection with non-subtype B strains. Analysis of gag p24, pol IN, and env gp41 IDR revealed the presence of five subtypes (A, B, C, D, H), CRF02_AG, and four unique recombinant forms. Due to the paucity of complete subtype H genomes available, we performed near full length genome sequence analysis on the candidate subtype H strain, designated as 00GB.AC4001. Phylogenetic analysis revealed that it formed a monophyletic cluster with the three available subtype H reference sequences. Bootscanning analysis confirmed that 00GB.AC4001 represents a new nonrecombinant subtype H genome. PMID- 19552592 TI - Analysis of HIV type 1 gp41 and enfuvirtide susceptibility among men in the United States who were HIV infected prior to availability of HIV entry inhibitors. AB - We analyzed HIV gp41 from 195 men in the United States who were HIV-1 infected between 1999 and 2002, before enfuvirtide (ENF) was approved for clinical use in the United States. gp41 genotyping results were obtained for 175 samples. None of the samples had major ENF resistance mutations. Six (3.4%) samples had minor ENF resistance mutations in the HR1 region (V38G, N43K, L44M, L45M). Twenty-eight (16%) samples had the N42S polymorphism, which is associated with ENF hypersusceptibility. Accessory mutations in the HR2 region were identified in some samples (E137K, S138A). Five of the six samples with HR1 resistance mutations were analyzed with a phenotypic assay; one sample had reduced ENF susceptibility (a sample with N42S +L44M + E137K). Prior to the availability of ENF, some men in the United States were infected with HIV that contained mutations associated with ENF resistance or hypersusceptibility. However, most of the mutations were not associated with phenotypic ENF resistance. PMID- 19552593 TI - In utero HIV infection is associated with an increased risk of nevirapine resistance in ugandan infants who were exposed to perinatal single dose nevirapine. AB - Use of single dose nevirapine (sdNVP) to prevent HIV mother-to-child transmission is associated with the emergence of NVP resistance in many infants who are HIV infected despite prophylaxis. We combined results from four clinical trials to analyze predictors of NVP resistance in sdNVP-exposed Ugandan infants. Samples were tested with the ViroSeq HIV Genotyping System and a sensitive point mutation assay (LigAmp, for detection of K103N, Y181C, and G190A). NVP resistance was detected at 6-8 weeks in 36 (45.0%) of 80 infants using ViroSeq and 33 (45.8%) of 72 infants using LigAmp. NVP resistance was more frequent among infants who were infected in utero than among infants who were diagnosed with HIV infection after birth by 6-8 weeks of age. Detection of NVP resistance at 6-8 weeks was not associated with HIV subtype (A vs. D), pre-NVP maternal viral load or CD4 cell count, infant viral load at 6-8 weeks, or infant sex. NVP resistance was still detected in some infants 6-12 months after sdNVP exposure. In this study, in utero HIV infection was the only factor associated with detection of NVP resistance in infants 6-8 weeks after sdNVP exposure. PMID- 19552595 TI - A genome-wide scan for the Sasang constitution in a Korean family suggests significant linkage at chromosomes 8q11.22-23 and 11q22.1-3. AB - OBJECTIVES: We aim to identify the genetic loci responsible for Sasang constitution type, which is important for effective personalized treatments in traditional Korean medicine. SUBJECTS: Forty (40) individuals in a Korean family were recruited for linkage analysis and 310 unrelated individuals for association analysis to confirm the linkage result. OUTCOME MEASURES: Genome-wide linkage analysis was performed for the Korean family using the Affymetrix 500K arrays. MERLIN software was used for multipoint nonparametric linkage (NPL) analysis. The significant candidate regions in linkage analysis were also investigated with association analysis of independent 310 individuals. RESULTS: Linkage analysis showed four significant peaks, 3q27.3, 8p11.21, 8q11.22-23, and 11q22.1-3, whose NPL Z scores are greater than 5.0. Among the significant loci, the 8q11.22-23 and 11q22.1-3 regions were supported by independent association analysis at the level of p < 0.05. CONCLUSIONS: The 8q11.22-23 and 11q22.1-3 regions were suggested as the candidate region for significant linkage to Sasang constitution. PMID- 19552596 TI - Complete recovery from tension-type headache through Kampo medicine. AB - OBJECTIVE: We administered Kampo medicine for tension-type headache with extremely effective results. The objective is to demonstrate the effect of Kampo medicine. SUBJECT: The case was 13 year-old-girl who had been suffering from headaches from the age of 11 years. As general and neurologic examinations (blood tests, urine analysis, head computed tomography, and electroencephalography) showed no abnormal findings, we diagnosed her as suffering from tension-type headache. Accompanying signs and symptoms included fatigue, shoulder stiffness, and a lot of stress in a private supplementary school and from ballet lessons. INTERVENTIONS AND OUTCOME: Kampo medicine was chosen to address the necessity for a comprehensive treatment. We prescribed saikokaryukotsuboreito, with clear positive effect. Although headaches re-appeared when she later forgot to take the formula, the symptom disappeared by taking it again. Treatment was discontinued after 11 months, and the headache has not recurred for 3 years. CONCLUSIONS: Kampo medicine was able to improve her overall symptom pattern, as Kampo formulas are selected not only by paying attention to the primary symptom (in this case headache) but also by checking the other characteristics (additional symptoms, constitution, etc). This case showed that Kampo treatment is an effective therapeutic option for tension-type headaches accompanied by various other symptoms. PMID- 19552597 TI - Acupuncture for chemotherapy-induced neutropenia in patients with gynecologic malignancies: a pilot randomized, sham-controlled clinical trial. AB - OBJECTIVES: The objective of this study was to investigate the effect of acupuncture administered during myelosuppressive chemotherapy on white blood cell (WBC) count and absolute neutrophil count (ANC) in patients with ovarian cancer. DESIGN: This study is a pilot, randomized, sham-controlled clinical trial. Patients received active acupuncture versus sham acupuncture while undergoing chemotherapy. A standardized acupuncture protocol was employed with manual and electrostimulation. The frequency of treatment was 2-3 times per week for a total of 10 sessions, starting 1 week before the second cycle of chemotherapy. SETTING: The setting was two outpatient academic centers for patients with cancer. SUBJECTS: Twenty-one (21) newly diagnosed and recurrent ovarian cancer patients were the subjects. OUTCOME MEASURES: WBC count, ANC, and plasma granulocyte colony-stimulating factor (G-CSF ) were assessed weekly. RESULTS: The median leukocyte value in the acupuncture arm at the first day of the third cycle of chemotherapy was significantly higher than in the control arm after adjusting for baseline value (8600 cells/microL, range: 4800-12,000 versus 4400 cell/microL, range: 2300-10,000) (p = 0.046). The incidence of grade 2-4 leukopenia was less in the acupuncture arm than in the sham arm (30% versus 90%; p = 0.02). However, the median leukocyte nadir, neutrophil nadir, and recovering ANC were all higher but not statistically significantly different (p = 0.116-0.16), after adjusting for baseline differences. There were no statistically significant differences in plasma G-CSF between the two groups. CONCLUSIONS: We observed clinically relevant trends of higher WBC values during one cycle of chemotherapy in patients with ovarian cancer, which suggests a potential myeloprotective effect of acupuncture. A larger trial is warranted to more definitively determine the efficacy of acupuncture on clinically important outcomes of chemotherapy-induced neutropenia. PMID- 19552598 TI - Potential protective effect of long-term therapy with Xuezhikang on left ventricular diastolic function in patients with essential hypertension. AB - OBJECTIVE: To investigate the benefits of long-term therapy with Xuezhikang, a cholestin extract, in combination with calcium channel blockers for improvement of left ventricular (LV) hypertrophy and function in patients with essential hypertension, as determined using echocardiography. DESIGN: Fifty-five (55) hypertensive patients with normal blood low-density lipoprotein cholesterol (LDL C) levels were randomly assigned to the Xuezhikang group (n = 28, 1200 mg/d of Xuezhikang) or the placebo group (n = 27, matched placebo). All of the patients were treated with extended-release nifedipine (20 mg twice daily). Thirty (30) normotensive subjects, matched for age and gender, were selected as a control group. Conventional echocardiography and tissue Doppler imaging were used to measure the left ventricle (LV) wall thickness and LV diastolic function at weeks 0, 24, and 72 during the period of observation. The serum levels of lipids, carboxy-terminal propeptide of procollagen type I (PIP), and C-reactive protein (CRP) were determined as well. RESULTS: The hypertensive patients had significantly elevated PIP and CRP levels in serum, increased LV wall thickness, and impaired LV diastolic function compared with the normotensive subjects (0.01 < p < 0.05). Compared with the placebo group, the transmitral flow velocities (E/A ratio) (1.11 +/- 0.36 versus 0.85 +/- 0.24, p < 0.01) and the myocardial motion velocities (Em/Am ratio) at the septal mitral annulus (0.90 +/- 0.19 versus 0.70 +/- 0.18, p < 0.05) and the lateral mitral annulus (1.06 +/- 0.20 versus 0.86 +/- 0.14, p < 0.01) were significantly increased, while there was no significant change in the LV wall thickness after 72 weeks of therapy with Xuezhikang. The serum levels of PIP (0.43 +/- 0.13 ng/mL versus 0.51 +/- 0.20 ng/mL, p < 0.05) and CRP (0.32 +/- 0.13 mg/L versus 0.40 +/-0.17 mg/L, p < 0.05) were significantly reduced compared to placebo treatment. There was no significant correlation between changes in LV diastolic function and blood pressure or lipid profile with Xuezhikang therapy. CONCLUSION: Long-term therapy with Xuezhikang improved LV diastolic function, probably mediated through antifibrotic and anti-inflammatory effects and independent of blood pressure and lipid profiles in patients with essential hypertension. PMID- 19552599 TI - Understanding the reliability of diagnostic variables in a Chinese Medicine examination. AB - The question of the objectivity of the clinical examination has been raised in relation to Western and non-Western medical systems. Western practitioners are often skeptical about Traditional Chinese Medicine (TCM), on the basis that its diagnostic variables and subcategories of disease appear subjective and not repeatable. We conducted a study investigating the reproducibility of individual diagnostic observations within three of the four diagnostic methods used in a TCM examination: inspection, palpation, and auscultation. Three TCM practitioners participated in the study, and examined 45 adults who had mild-to-moderate hypercholesterolemia but were otherwise healthy. Results indicated that while there are certain features of the TCM system that are highly objective and repeatable, such as detection of the presence of shen, character of breath sounds, and pulse speed, there are other features that are subjective and unreliable, such as color under the eyes and tongue body color. This poses a challenge for TCM practitioners to improve their clinical practice and demonstrates to Western medical practitioners that TCM does in part rest on a rigorous and objective empirical basis. PMID- 19552600 TI - Effects of timolol and latanoprost on respiratory and cardiovascular status in elderly patients with glaucoma. AB - PURPOSE: To investigate the respiratory and cardiovascular effects of switching therapy from topical timolol 0.5% to latanoprost 0.005% in elderly patients with glaucoma. METHODS: Twenty-five patients with primary open-angle glaucoma (POAG) who were treated with topical timolol 0.5% were included in the study. The beta blocker treatment was ended due to insufficient intraocular pressure (IOP) and systemic or local side effects. The treatment then switched to latanoprost in these patients. All recruited patients underwent a full ocular, cardiovascular, and respiratory examination including spirometry, pulse rate, and blood pressure. All measurements were made 1 day before the beta-blocker treatment was ended and 30 days after the latanoprost treatment was started. RESULTS: Timolol treatment was associated with numerically but not statistically significant lowered pulse rates, systemic blood pressure, diastolic blood pressure, and reductions in the spirometry measurements. The change to latanoprost treatment was associated with numerical improvement in mean values of spirometric test results. There were no significant differences in changes in mean values of spirometry, pulse rate, or blood pressure. Histamine challenge test was determined to be positive in 16 of 25 patients for timolol, of whom 10 were positive for latanoprost. This was statistically significant (P < 0.05). CONCLUSIONS: Although all the results presented were statistically insignificant, switching to latanoprost treatment may offer some advantages in respiratory and cardiovascular function for elderly people with glaucoma over beta-blocker drugs. Spirometry and the measurement of pulse rate and blood pressure are advised in all patients receiving topical beta blocker therapy. PMID- 19552601 TI - Nepafenac dosing frequency for ocular pain and inflammation associated with cataract surgery. PMID- 19552594 TI - Comparison of HIV type 1 sequences from plasma, cell-free breast milk, and cell associated breast milk viral populations in treated and untreated women in Mozambique. AB - We analyzed the sequences of the HIV viral populations obtained from plasma, cell free breast milk, and breast milk cells of HAART-treated (23) and untreated (30) HIV-infected women to obtain information about the origin of the breast milk virus. Sequence analyses of viruses were performed using the TruGene HIV-1 assay. Direct sequences of the reverse transcriptase (RT) and protease (PR) genes were analyzed using the Phylip 3.68 suite of sequence analysis program and pairwise evolutionary distances were calculated with the Kimura two parameter model for estimation of distances. We found that the genetic distances between the plasma and the cell-free breast milk viruses and between the cell-free and cell associated breast milk viruses for RT were higher in HAART-receiving women than in untreated women, suggesting viral evolution under selective drug pressure in breast milk. Our data support the hypothesis of the presence of an actively replicating viral population in the breast milk compartment, distinct from that present in plasma. PMID- 19552602 TI - Activation of the BK(Ca) channel increases outflow facility and decreases trabecular meshwork cell volume. AB - PURPOSE: Inhibition of the BK(Ca) channel attenuated the nitric oxide-induced increase in outflow facility and decrease in trabecular meshwork (TM) cell volume suggesting the involvement of the BK(Ca) channel in TM cell function. This study examined the effects of activation of the BK(Ca) channel on outflow facility and TM cell volume and determined if the effects of NO and BK(Ca) channel activation on TM cell volume were additive. METHODS: Porcine eyes were used to measure outflow facility using the anterior segment organ culture perfusion system. Cell volume was measured using Calcein AM fluorescent dye, detected by confocal microscopy, and quantified using NIH ImageJ software. RESULTS: NS1619 increased outflow facility 86% over baseline. Additionally, there was a concentration dependent decrease in TM cell volume in response to NS1619, which was abolished by iberiotoxin (IBTX). While NS1619 alone and DETA-NO alone decreased TM cell volume, together their effects were not additive. The time course for NS1619 induced increases in outflow facility correlated with the time course for NS1619 induced decreases in cell volume. CONCLUSIONS: BK(Ca) channel activation increases outflow facility and decreases cell volume suggesting that K(+) efflux regulates TM cell function. PMID- 19552603 TI - Architectural and immunohistochemical characterization of biliary ductules in normal human liver. AB - The canals of Hering or biliary ductules have been described to connect the bile canaliculi with the interlobular bile ducts, and thus forming the distal part of the biliary tree. Studies in the last two decades suggested that the cells constructing these ductules could behave as hepatic progenitor cells. The canals of Hering are confined to the periportal space in the rat, while they have been reported to spread beyond the limiting plate in human liver. The distribution of the distal biliary ductules in normal human hepatic tissue has been investigated in our recent experiments. We could demonstrate the presence of interlobular connective tissue septa in a rudimentary form in healthy livers. The canals of Hering run in these septa in line with the terminal branches of the portal vein and hepatic arteries. This arrangement develops in the postnatal period but regresses after early childhood. The canals of Hering can be identified by the unique epithelial membrane antigen (EMA)-/CD56+/CD133+ immunophenotype. The canals of Hering leave the periportal space and spread into the liver parenchyma along rudimentary interlobular septa outlining the hepatic lobules. Our observations refine the original architectural description of the intraparenchymal portion of the canals of Hering in the human liver. The distinct immunophenotype supports their unique biological function. PMID- 19552605 TI - Inosculation: connecting the life-sustaining pipelines. AB - Recent progress in engineering microvascular networks in vitro and in vivo offers exciting opportunities to create tissue constructs with preformed blood vessels, which are rapidly blood perfused by developing interconnections to the preexisting blood vessels of the host tissue after implantation. This process, termed as inosculation, is well known from the revascularization of various tissue grafts, such as transplanted skin, nerves, or bone. It is characterized by the close interaction of the implant's preformed microvascular network and the host microvasculature. The sprouting angiogenic activity of both counterparts determines whether inosculation takes place internally within the implant or externally within the surrounding host tissue. Successful inosculation involves vascular remodeling as well as infiltration of inflammatory cells and stem cells. With the use of sophisticated in vitro and in vivo models, more detailed analysis of regulatory mechanisms of inosculation will help to develop novel strategies, aiming at further accelerating the establishment of a life-sustaining blood supply to implanted tissue constructs. PMID- 19552604 TI - Micro- and nanoscale control of the cardiac stem cell niche for tissue fabrication. AB - Advances in stem cell (SC) biology have greatly enhanced our understanding of SC self-renewal and differentiation. Both embryonic and adult SCs can be differentiated into a great variety of tissue cell types, including cardiac myocytes. In vivo studies and clinical trials, however, have demonstrated major limitations in reconstituting the myocardium in failing hearts. These limitations include precise control of SC proliferation, survival and phenotype both prior and subsequent to transplantation and avoidance of serious adverse effects such as tumorigenesis and arrhythmias. Micro- and nanoscale techniques to recreate SC niches, the natural environment for the maintenance and regulation of SCs, have enabled the elucidation of novel SC behaviors and offer great promise in the fabrication of cardiac tissue constructs. The ability to precisely manipulate the interface between biopolymeric scaffolds and SCs at in vivo scale resolutions is unique to micro- and nanoscale approaches and may help overcome limitations of conventional biological scaffolds and methods for cell delivery. We now know that micro- and nanoscale manipulation of scaffold composition, mechanical properties, and three-dimensional architecture have profound influences on SC fate and will likely prove important in developing the next generation of "transplantable SC niches" for regeneration of heart and other tissues. In this review, we examine two key aspects of micro- and nanofabricated SC-based cardiac tissue constructs: the role of scaffold composition and the role of scaffold architecture and detail how recent work in these areas brings us closer to clinical solutions for cardiovascular regeneration. PMID- 19552607 TI - Emerging drugs for hepatic encephalopathy. AB - BACKGROUND: Hepatic encephalopathy (HE) is a severe complication of cirrhosis, seriously affecting the patients' quality of life. The classical approach aimed at reducing the production of gut-derived toxins, such as ammonia, is under debate as, at the moment, the information obtained from the clinical trials does not support any specific treatment for HE. OBJECTIVES: i) To discuss present therapeutic strategies and possible future developments; ii) to identify areas of medical needs and iii) to suggest the ideal design and methodology for randomized controlled trials (RCTs) in HE. METHODS: Current approaches were obtained from already available RCTs or from experimental animal studies. Those approaches developed from studies on HE pathophysiology were considered as working hypotheses for future therapies. RESULTS/CONCLUSION: Our competence in testing old and new treatment modalities by RCTs with appropriate clinically relevant end points should urgently be improved. The patients at risk of HE are identifiable, and studies specifically aimed at establishing whether HE may be prevented or not are needed. As far as new treatment modalities are concerned, RCTs on the modulators of the intestinal bacterial flora and on the molecular adsorbent recirculating system are already available, but further studies are needed to confirm these promising approaches. PMID- 19552606 TI - Endogenous adenosine selectively modulates oxidant stress via the A1 receptor in ischemic hearts. AB - We tested the impact of A1 adenosine receptor (AR) deletion on injury and oxidant damage in mouse hearts subjected to 25-min ischemia/45-min reperfusion (I/R). Wild-type hearts recovered approximately 50% of contractile function and released 8.2 +/- 0.7 IU/g of lactate dehydrogenase (LDH). A1AR deletion worsened dysfunction and LDH efflux (15.2 +/- 2.6 IU/g). Tissue cholesterol and native cholesteryl esters were unchanged, whereas cholesteryl ester-derived lipid hydroperoxides and hydroxides (CE-O(O)H; a marker of lipid oxidation) increased threefold, and alpha-tocopherylquinone [alpha-TQ; oxidation product of alpha tocopherol (alpha-TOH)] increased sixfold. Elevations in alpha-TQ were augmented by two- to threefold by A1AR deletion, whereas CE-O(O)H was unaltered. A(1)AR deletion also decreased glutathione redox status ([GSH]/[GSSG + GSH]) and enhanced expression of the antioxidant response element heme oxygenase-1 (HO-1) during I/R: fourfold elevations in HO-1 mRNA and activity were doubled by A1AR deletion. Broad-spectrum AR agonism (10 microM 2-chloroadenosine; 2-CAD) countered effects of A1AR deletion on oxidant damage, HO-1, and tissue injury, indicating that additional ARs (A(2A), A(2B), and/or A3) can mediate similar actions. These data reveal that local adenosine engages A1ARs during I/R to limit oxidant damage and enhance outcome selectively. Control of alpha-TOH/alpha-TQ levels may contribute to A1AR-dependent cardioprotection. PMID- 19552608 TI - Emerging drugs for the therapy of primary and post essential thrombocythemia, post polycythemia vera myelofibrosis. AB - Managing patients with myelofibrosis (MF) (both those with primary myelofibrosis or having evolved from an antecedent polycythemia vera or essential thrombocythemia) present many challenges to the hematologist. MF patients suffer from a variable, but severe range of disease manifestations including massive splenomegaly, cytopenias, significant constitutional symptoms, possible transformation to blast phase and premature death. Cure is achievable through allogeneic stem cell transplantation; yet, this therapy is either inappropriate or not an option for most patients. Current available therapies are palliative, but can sometimes be of significant value to MF patients. The discovery of the JAK2-V617F mutation, and other pathogenetic insights into the pathophysiology of myeloproliferative neoplasms, has ushered in an era of potential new therapies for MF. Over a dozen JAK2 inhibitors are in development, with the leading compounds such as INCB018424, TG101348 and others showing promising early results particularly for control of disease associated splenomegaly and symptoms. Parallel trials with immunomodulatory therapy for MF associated anemia and stromal manifestations of the disease are continuing. The future may well see the approval of a range of agents for MF patients, with differing mechanisms of action, efficacy and toxicity profiles. PMID- 19552609 TI - Malignant pleural mesothelioma: current treatments and emerging drugs. AB - BACKGROUND: Malignant pleural mesothelioma (MPM) is an uncommon disease whose incidence is increasing worldwide over the past 30 years. Surgical resection and radiotherapy represent the standard treatment in patient with resectable MPM. Chemotherapy is also necessary to reduce incidence of distant metastases, but the optimal setting of treatment (neoadjuvant, adjuvant and intrapleural) is not clarified. For the patients with unresectable MPM, the combination cisplatin and pemetrexed or ralitrexed is the standard treatment as supported by a Phase III study. Better understanding of molecular pathways involved in MPM has enabled inclusion of new drugs targeted against pathways responsible for proliferation, cell survival and angiogenesis. OBJECTIVE: This review discusses the current treatment option, the specific signal pathways activated in MPM and the novel agents under evaluation in clinical trials. METHODS: We use for this article abstracts, papers, oral presentations from ASCO and the website http://www.clinical-trials.gov. RESULTS/ CONCLUSION: This review summarizes the activity of chemotherapy and of new agents under evaluation in clinical trials. The better understanding of molecular pathways activated in MPM will hopefully provide new therapeutic options for these patients in the future. PMID- 19552610 TI - Drugs with anticholinergic properties: a potential risk factor for psychosis onset in Alzheimer's disease? AB - Psychosis in Alzheimer's disease is common and troublesome. The impact on the quality of life of both patients and caregivers is high and drug treatments raise concern in terms of both efficacy and safety. Therefore, identifying the risk factors that play an important role in the onset of psychosis is mandatory for the prevention of this clinical condition. From a biological point of view, drugs with anticholinergic properties are a reasonable cause of psychosis. Demented patients have been found to use a disproportionate amount of drugs with anticholinergic properties. On the other hand, new evidence suggests that the cholinergic system may be implicated not only with the onset of cognitive impairment, but even in the genesis of psychosis symptoms. This review focuses on biological and clinical data which suggest that anti-cholinergic drugs should be regarded as a potential risk factor for psychosis in Alzheimer's disease. PMID- 19552612 TI - Collagen-coated microparticles in drug delivery. AB - Advantages of drug-incorporated collagen particles have been described for the controlled delivery system for therapeutic actions. The attractiveness of collagen lies in its low immunogenicity and high biocompatibility. It is also recognized by the body as a natural constituent rather than a foreign body. Our research and development efforts are focused towards addressing some of the limitations of collagen, like the high viscosity of an aqueous phase, nondissolution in neutral pH buffers, thermal instability (denaturation) and biodegradability, to make it an ideal material for drug delivery with particular reference to microparticles. These limitations could be overcome by making collagen conjugates with other biomaterials or chemically modifying collagen monomer without affecting its triple helical conformation and maintaining its native properties. This article highlights collagen microparticles' present status as a carrier in drug delivery. PMID- 19552611 TI - Controlled delivery of antisense oligonucleotides: a brief review of current strategies. AB - Antisense therapy has been investigated extensively over the past two decades, either experimentally for gene functional research or clinically as therapeutic agents owing to the conceptual simplicity, ease of design and low cost. The concept of this therapeutic approach is promising because short antisense oligonucleotides (ASOs) can be delivered into target cells for specific hybridisation with target mRNA, resulting in the inhibition of the expression of pathogenic genes. However, the efficient delivery of the ASO molecules into target cells remains challenging; this bottleneck together with several other technical hurdles need to be overcome before this approach becomes effective and widely adopted. A variety of vectors such as lipids, polymers, peptides and nanoparticles have been explored. This review outlines the recent advances of the non-viral ASO delivery strategies. Several recent scientific studies, including authors' contributions, have been selected to highlight the technical aspects of ASO delivery. PMID- 19552613 TI - Nuclear targeting of viral and non-viral DNA. AB - The nuclear envelope presents a major barrier to transgene delivery and expression using a non-viral vector. Virus is capable of overcoming the barrier to deliver their genetic materials efficiently into the nucleus by virtue of the specialized protein components with the unique amino acid sequences recognizing cellular nuclear transport machinery. However, considering the safety issues in the clinical gene therapy for treating critical human diseases, non-viral systems are highly promising compared with their viral counterparts. This review summarizes the progress on exploring the nuclear traffic mechanisms for the prominent viral vectors and the technological innovations for the nuclear delivery of non-viral DNA by mimicking those natural processes evolved for the viruses as well as for many cellular proteins. PMID- 19552614 TI - Applications for site-directed molecular imaging agents coupled with drug delivery potential. AB - Molecular imaging allows non-invasive characterization and quantification of biological processes at cellular and molecular level. Such technologies make it possible to enhance our understanding of drug activity and pharmokinetic properties, and therefore aid decisions to select candidates that are most likely to benefit from targeted drug therapy. Targeted DDSs are nanometer-sized carrier materials designed for improving the biodistribution of systemically applied (chemo-)therapeutics by strictly localizing its pharmacological activity to the site or organ of action. The parallel development of molecular imaging and targeted drug delivery offers great challenges and opportunities for a single multifunctional platform technology, combining targeted motif, therapeutic agents and imaging agents for imaging guided drug delivery. This review article summarizes the synthesis and characterization of various biomaterials that carry targeting motifs, imaging tags and therapeutic agents as theragnostics. PMID- 19552615 TI - Revealing evolutionary pathways by fitness landscape reconstruction. AB - The concept of epistasis has since long been used to denote non-additive fitness effects of genetic changes and has played a central role in understanding the evolution of biological systems. Owing to an array of novel experimental methodologies, it has become possible to experimentally determine epistatic interactions as well as more elaborate genotype-fitness maps. These data have opened up the investigation of a host of long-standing questions in evolutionary biology, such as the ruggedness of fitness landscapes and the accessibility of mutational trajectories, the evolution of sex, and the origin of robustness and modularity. Here we review this recent and timely marriage between systems biology and evolutionary biology, which holds the promise to understand evolutionary dynamics in a more mechanistic and predictive manner. PMID- 19552617 TI - Metabonomic characterization of early atherosclerosis in hamsters with induced cholesterol. AB - Atherosclerosis is a complicated and multifactorial disease, induced not only by genotype, but also, even more importantly, by environmental factors. Study on the metabolic perturbation of endogenous compounds may offer deeper insight into development of atherosclerosis. Gas chromatography/mass spectrometry (GC/MS) based metabonomics was used to profile a metabolic fingerprint of serum obtained from hamsters with induced cholesterol. The deconvoluted GC/MS data were processed by multivariate statistical analysis tools, such as principal component analysis (PCA) and partial least squares projection to latent structure and discriminant analysis (PLS-DA). For the first time we showed a time-dependent development of the model animal from normal to hypercholesterolaemia, and further to early atherosclerosis. Twenty-one compounds were identified as markers involved in the development to atherosclerosis. Identification of the compounds suggests that amino acid metabolism and fatty acid oxidation are significantly perturbed following cholesterol overloading. The data provide novel information to approach the pathophysiological processes of the hypercholesterolaemia and atherosclerosis disease continuum. PMID- 19552616 TI - Lung epithelium injury biomarkers in workers exposed to sulphur dioxide in a non ferrous smelter. AB - Serum Clara cell protein (CC16) and surfactant-associated protein D (SP-D) were measured in 161 workers exposed to sulphur dioxide (SO(2)) in a non-ferrous smelter. Seventy workers from a blanket manufacture served as referents. Exposure to SO(2) and tobacco smoking were associated with a decrease of CC16 and an increase of SP-D in serum. Tobacco smoking and exposure SO(2) interacted synergistically to decrease serum CC16 but not to increase serum SP-D. While further illustrating the potential of serum CC16 and SP-D, our study confirms that SO(2) can cause airways damage at exposure levels below current occupational exposure limits. PMID- 19552618 TI - Acute conjunctivitis: truth and misconceptions. AB - OBJECTIVE: Acute conjunctivitis is an extremely common condition and can be associated with significant morbidity and economic burden. Despite this, considerable controversy exists pertaining to the prevalence, diagnosis, management, and treatment of the condition. A panel of ophthalmology experts was assembled to review and discuss the current evidence based literature as it pertains to each of these persistent controversies. METHODS AND SCOPE: An acute conjunctivitis round table symposium was convened at the American Academy of Ophthalmology meeting in Atlanta, November 2008. The expert panelists consisted of four academic ophthalmologists in the field of cornea and external disease, whose discussion was informed by an English language literature survey carried out on the PubMed database for the period of January 1972 to October 2008. A narrative summary was generated from the literature review and direct transcription of this event, from which this Review article was developed. FINDINGS AND CONCLUSIONS: Considerable light has been shed on acute microbial conjunctivitis and especially those cases caused by adenovirus. Many of the myths that have perpetuated for years have been debunked by emerging evidence. The advent and the implementation of better diagnostic tools and anti-viral medications will help clinicians to improve their diagnostic accuracy, improve management and treatment decisions, and ultimately benefit patients while saving overall healthcare costs. PMID- 19552619 TI - Evaluation of the potential for steady-state pharmacokinetic and pharmacodynamic interactions between the DPP-4 inhibitor linagliptin and metformin in healthy subjects. AB - OBJECTIVE: Linagliptin (BI 1356) is a novel, orally available inhibitor of dipeptidyl peptidase-4 (DPP-4). Linagliptin improves glycaemic control in type 2 diabetic patients by increasing the half-life of the incretin hormone glucagon like peptide-1 (GLP-1). Linagliptin is expected to be used as monotherapy or in combination with other antihyperglycaemic agents. This study was conducted to investigate potential pharmacokinetic or pharmacodynamic interactions between linagliptin and metformin. METHODS: This randomised, monocentric, open-label, two way crossover design study was conducted in 16 healthy male subjects. Linagliptin (10 mg/day) and metformin (850 mg three times daily) were each administered alone and concomitantly. The steady-state pharmacokinetics of linagliptin and metformin and the inhibition of DPP-4 activity were determined at the end of each dosing period. RESULTS: Co-administration of linagliptin had no apparent effect on metformin exposure (metformin AUC(tau,ss); geometric mean ratio [GMR] co administration:individual administration was 1.01; 90% confidence interval [CI] was 0.89-1.14). Effects on maximum concentration (C(max,ss)) were small (GMR: 0.89; 90% CI: 0.78-1.00). Co-administration of metformin did not significantly affect C(max,ss) of linagliptin (GMR: 1.03; 90% CI: 0.86-1.24), but increased AUC(tau)(,ss) by 20% (GMR: 1.20; 90% CI: 1.07-1.34). Metformin alone had no effect on DPP-4 activity, and the inhibition of DPP-4 caused by linagliptin was not affected by concomitant administration of metformin. Tolerability was good whether linagliptin and metformin were administered alone or concomitantly. No serious adverse events occurred and the frequency of adverse events was low; 7 events in 6 subjects. The most frequent events were related to the gastrointestinal tract, as expected with metformin. Importantly, no subjects experienced signs or symptoms relating to episodes of hypoglycaemia. CONCLUSION: In this small, multiple dose study carried out in healthy subjects, co administration of linagliptin with metformin did not have a clinically relevant effect on the pharmacokinetics or pharmacodynamics of either agent. This study suggests linagliptin and metformin can safely be administered concomitantly in type 2 diabetes patients without dose adjustment; larger, longer-term clinical trials in diabetic patients are underway. PMID- 19552620 TI - Estimation of economic costs associated with transfusion dependence in adults with MDS. AB - OBJECTIVE: To examine the economic burden of myelodysplastic syndromes (MDS) and the incremental cost of transfusion dependence. RESEARCH DESIGN AND METHODS: Adults with evidence of MDS were identified between 05/01/2000 and 09/30/2003 from a longitudinal, retrospective claims database for a large, geographically diverse US health plan and their medical histories were followed for at least 6 months. Patients were classified as transfusion-dependent (MDS-TD) or transfusion independent (MDS-TI). MAIN OUTCOME MEASURES: Variables were categorized as demographic, health status, utilization, or cost. Utilization (inpatient hospitalizations, outpatient facility visits, emergency department visits, and physician office visits) is reported as the mean and median numbers of each specified encounter per subject. Costs were measured as the sum of patient and plan liability. All variables were analyzed descriptively, and appropriate statistical tests were used to compare the MDS-TD and MDS-TI cohorts. Pharmacy, medical, and total health care costs, adjusted for demographics and comorbidity, were estimated using gamma regression with a log link. RESULTS: The MDS-TI cohort consisted of 2864 patients, and the MDS-TD cohort comprised 336 patients. Mean age for the entire study sample was 70.2 years. The MDS-TI cohort tended to receive most of its medical care at physicians' offices, whereas the MDS-TD cohort received nearly as much medical care at outpatient facilities (e.g., infusion clinics, hospital outpatient clinics) as it did in physicians' offices. The MDS-TD cohort had significantly higher mean annual costs: pharmacy, $4457 vs. $2926; medical, $50,663 vs. $17,469; total, $51,066 vs. $19,811 (p < 0.001 for all comparisons). Thus, transfusion dependence was associated with an incremental cost of $31,255 per patient per year. Some limitations inherent to using claims data and diagnosis codes for research apply to this study. CONCLUSIONS: This study demonstrated that an important consequence of transfusion dependence for MDS patients was markedly greater use of, and consequently higher costs associated with, inpatient and outpatient services. Continued research and efforts to develop biologic and pharmaceutical therapies may help more patients achieve transfusion independence, thereby reducing the financial burden of MDS. PMID- 19552621 TI - A novel application for Cocoacrisp protein as a biomarker for experimental pulmonary fibrosis. AB - Pulmonary fibrosis is a debilitating disease affecting up to 2 million people worldwide, with a median survival rate of only 3 years after diagnosis. The aim of this study was to evaluate a potential protein biomarker (Cocoacrisp, CC) to identify the onset of pulmonary fibrosis. A model of fibrosis was induced via intratracheal instillation of bleomycin, and samples were collected during the early phase of the disease. Immunohistochemical identification of CC was carried out in lung tissue from the bleomycin model. Quantification by image analysis showed CC levels were doubled (p <0.0003), after a single bleomycin dose, but not after double instillation. Microscopic analysis revealed that CC signal was primarily detected on the alveolar surface. The secretion of the novel protein CC during the early stages of bleomycin-induced injury may have the potential to be utilized as a clinical biomarker for the early stages of fibrosis, particularly as it may be detectable in bronchoalveolar lavage fluid. PMID- 19552622 TI - Proteomic analysis of mouse thymoma EL4 cells treated with bis(tri-n butyltin)oxide (TBTO). AB - Here, we report the results of proteomic analysis of the mouse thymoma EL4 cell line exposed to bis(tri-n-butylin)oxide (TBTO), an immunotoxic organotin compound. The objective of the work was to examine whether TBTO affects the expression of proteins in this cell line and to compare the differentially expressed proteins with the corresponding mRNA expression data. The identified proteins were quantified using a label-free quantitative method based on counting the observed peptides as an index of protein abundance. The calculation of the ratio of peptides obtained from exposed and control samples allowed us to evaluate the effect of TBTO on protein expression and to compare these results to those obtained in gene expression profiling studies. Correlation of some of the differentially expressed proteins and their corresponding mRNAs was observed. The analysis of the protein ratios revealed that 12 proteins were significantly affected. These proteins included cytoskeleton proteins myosin-9, spectrin beta 2 and plectin 8. The first two proteins were down-regulated 3-fold, whereas the third was up-regulated 2-fold. Ras-related Rab1, a GTP binding protein and T complex protein-1 subunit alpha, a chaperonin, were decreased 2- and 3.6-fold, respectively. The ribosomal S10 and eukaryotic translation factor (eIf4G1), which are involved in protein synthesis, were down-regulated 2.6- and 3.7-fold, respectively. Also, proteins involved in splicing of pre-mRNA and in transcription, splicing factor arginine/serine-rich 2 and chromodomain-helicase DNA binding protein 4 (Chd4), were decreased 2.6- and 4.5 times, respectively. Nuclear RNA helicase II was reduced 2.8-fold. Finally, prothymosin-alpha (ProTalpha), an essential protein for cell proliferation, and a protein similar to ProTalpha, (with a molecular weight and a pI (3.54) comparable to that of ProTalpha) were also down-regulated 6-and 8-fold, respectively. We propose that the observed down-regulation of the expression level of ProTalpha in the TBTO exposed cells could account for the previously reported anti-proliferative effect of TBTO. PMID- 19552623 TI - Pancreatic stellate cells can form new beta-like cells. AB - Regenerative medicine, including cell-replacement strategies, may have an important role in the treatment of Type 1 and Type 2 diabetes, both of which are associated with decreased islet cell mass. To date, significant progress has been made in deriving insulin-secreting beta-like cells from human ES (embryonic stem) cells. However, the cells are not fully differentiated, and there is a long way to go before they could be used as a replenishable supply of insulin-secreting beta-cells for transplantation. For this reason, adult pancreatic stem cells are seen as an alternative source that could be expanded and differentiated ex vivo, or induced to form new islets in situ. In this issue of the Biochemical Journal, Mato et al. used drug selection to purify a population of stellate cells from explant cultures of pancreas from lactating rats. The selected cells express some stem-cell markers and can be grown for over 2 years as a fibroblast-like monolayer. When plated on extracellular matrix, along with a cocktail of growth factors that included insulin, transferrin, selenium and the GLP-1 (glucagon-like peptide-1) analogue exendin-4, the cells differentiated into cells that expressed many of the phenotypic markers characteristic of a beta-cell, and exhibited an insulin-secretory response, albeit weak, to glucose. The ability to purify this cell population opens up the possibility of unravelling the mechanisms that control self-renewal and differentiation of pancreatic cells that share some of the properties of stem cells. PMID- 19552625 TI - Identification and functional characterization of legumain in amphioxus Branchiostoma belcheri. AB - Legumain has been reported from diverse sources such as plants, parasites (animals) and mammals, but little is known in the lower chordates. The present study reports the first characterization of legumain cDNA from the protochordate Branchiostoma belcheri. The deduced 435-amino-acid-long protein is structurally characterized by the presence of a putative N-terminal signal peptide, a peptidase_C13 superfamily domain with the conserved Lys123-Gly124-Asp125 motif and catalytic dyad His153 and Cys195 and two potential Asn-glycosylation sites at Asn85 and Asn270. Phylogenetic analysis demonstrates that B. belcheri legumain forms an independent cluster together with ascidian legumain, and is positioned at the base of vertebrate legumains, suggesting that B. belcheri legumain gene may represent the archetype of vertebrate legumain genes. Both recombinant legumain expressed in yeast and endogenous legumain are able to be converted into active protein of approximately 37 kDa via a C-terminal autocleavage at acid pH values. The recombinant legumain efficiently degrades the legumain-specific substrate Z-Ala-Ala-Asn-MCA (benzyloxycarbonyl-L-alanyl-L-alanyl-L-asparagine-4 methylcoumaryl-7-amide) at optimum pH 5.5; and the enzymatic activity is inhibited potently by iodoacetamide and N-ethylmaleimide, partially by hen's-egg white cystatin, but not by E-64 [trans-epoxysuccinyl-L-leucylamido-(4 guanidino)butane], PMSF and pepstatin A. In addition, legumain is expressed in vivo in a tissue-specific manner, with main expression in the hepatic caecum and hind-gut of B. belcheri. Altogether, these results suggest that B. belcheri legumain plays a role in the degradation of macromolecules in food. PMID- 19552626 TI - Chemokine degradation by the Group A streptococcal serine proteinase ScpC can be reconstituted in vitro and requires two separate domains. AB - Streptococcus pyogenes is one of the most common human pathogens and possesses diverse mechanisms to evade the human immune defence. One example of its immune evasion is the degradation of the chemokine IL (interleukin)-8 by ScpC, a serine proteinase that prevents the recruitment of neutrophils to an infection site. By applying the ANTIGENome technology and using human serum antibodies, we identified Spy0416, annotated as ScpC, as a prominent antigen that induces protective immune responses in animals. We demonstrate here for the first time that the recombinant form of Spy0416 is capable of IL-8 degradation in vitro in a concentration- and time-dependent manner. Mutations in the conserved amino acid residues of the catalytic triad of Spy0416 completely abolished in vitro activity. However, the isolated predicted proteinase domain does not exhibit IL-8 degrading activity, but is dependent on the presence of the C-terminal region of Spy0416. Binding to IL-8 is mainly mediated by the catalytic domain. However, the C-terminal region modulates substrate binding, indicating that the proteolytic activity is amenable to regulation via the non-catalytic regions. The specificity for human substrates is not restricted to IL-8, since we also detected in vitro protease activity for another CXC chemokine GRO-alpha (growth-related oncogene alpha), but not for NAP-2 (neutrophil-activating protein 2), SDF (stromal-cell derived factor)-1alpha, PF-4 (platelet factor 4), I-TAC (interferon-gamma inducible T-cell alpha-chemoattractant), IP-10 (interferon-gamma-inducible protein 10) and MCP-1 (monocyte chemoattractant protein 1). The degradation of two human CXC chemokines in vitro, the high sequence conservation, the immunogenicity of the protein in humans and the shown protection in animal studies suggest that Spy0416 is a promising vaccine candidate for the prevention of infections by S. pyogenes. PMID- 19552624 TI - Function and regulation of phospholipid signalling in plants. AB - As an important metabolic pathway, phosphatidylinositol metabolism generates both constitutive and signalling molecules that are crucial for plant growth and development. Recent studies using genetic and molecular approaches reveal the important roles of phospholipid molecules and signalling in multiple processes of higher plants, including root growth, pollen and vascular development, hormone effects and cell responses to environmental stimuli plants. The present review summarizes the current progress in our understanding of the functional mechanism of phospholipid signalling, with an emphasis on the regulation of Ins(1,4,5)P3 Ca2+ oscillation, the second messenger molecule phosphatidic acid and the cytoskeleton. PMID- 19552628 TI - An overview of osteoporosis for the practising prosthodontist. AB - Osteoporosis is an insidious and common bone disorder of the modern age, as a result of the rapidly increasing number of older people in the total population. It has long been concluded that this disease has definite deleterious effects on the stomatognathic system and is therefore of major concern to a Prosthodontist. If features on a dental radiograph, which are the most commonly required radiographs, can be detected regularly and consistently, it would place a prosthodontist in a position to refer the patient for timely management and also modify his treatment plan, greatly improving the prognosis. Available literature was therefore reviewed for pathophysiology, dental radiographic screening measures, implications and management of osteoporosis from the perspective of a prosthodontist. PMID- 19552629 TI - Screening of edentulous patients in a dental school population using the prosthodontic diagnostic index. AB - PURPOSE: The aim of this study was to classify a number of completely or partially edentulous patients who attended undergraduate clinics at the Dental School of Athens using the prosthodontic diagnostic index (PDI) to identify the complexity of the cases treated by the students. BACKGROUND: There is a scarcity of studies, concerning prosthodontic patients and screening them according to a classification system such as the PDI. MATERIAL AND METHODS: The survey was conducted on 181 patients who attended the clinics for treatment. The PDI categorised the patients into four classes (Class I-IV) according to the severity of certain diagnostic findings. RESULTS: From the 110 completely edentulous patients, 27 patients were in Class I, 21 in Class II, 15 in Class III and 47 in Class IV. From the 71 partially edentulous patients, two were in Class I, 17 in Class II, 29 in Class III and 23 in Class IV. CONCLUSION: Our results showed that the majority of patients were categorised in Class III (partially edentulous) and IV (completely edentulous). This indicates the need to introduce a proper screening tool, such as the PDIs, during the initial examination, to achieve a successful treatment. PMID- 19552627 TI - Ephrin-independent regulation of cell substrate adhesion by the EphB4 receptor. AB - Receptor tyrosine kinases of the Eph family become tyrosine phosphorylated and initiate signalling events upon binding of their ligands, the ephrins. Eph receptors such as EphA2 and EphB4 are highly expressed but poorly tyrosine phosphorylated in many types of cancer cells, suggesting a limited interaction with ephrin ligands. Nevertheless, decreasing the expression of these receptors affects the malignant properties of cancer cells, suggesting that Eph receptors may influence cancer cells independently of ephrin stimulation. Ligand independent activities of Eph receptors in cancer, however, have not been demonstrated. By using siRNA (small interfering RNA) to downregulate EphB4 in MCF7 and MDA-MB-435 cancer cells, we found that EphB4 inhibits integrin-mediated cell substrate adhesion, spreading and migration, and reduces beta1-integrin protein levels. Low expression of the EphB4 preferred ligand, ephrin-B2, and minimal contact between cells in these assays suggest that cell contact-dependent stimulation of EphB4 by the transmembrane ephrin-B2 ligand does not play a role in these effects. Indeed, inhibitors of ephrin-B2 binding to endogenous EphB4 did not influence cell substrate adhesion. Increasing EphB4 expression by transient transfection inhibited cell substrate adhesion, and this effect was also independent of ephrin stimulation because it was not affected by single amino acid mutations in EphB4 that impair ephrin binding. The overexpressed EphB4 was tyrosine phosphorylated, and we found that EphB4 kinase activity is important for inhibition of integrin-mediated adhesion, although several EphB4 tyrosine phosphorylation sites are dispensable. These findings demonstrate that EphB4 can affect cancer cell behaviour in an ephrin-independent manner. PMID- 19552630 TI - Transient lower oesophageal sphincter relaxation in achalasia: everything but LOS relaxation. AB - In conducting clinical high-resolution oesophageal pressure topography (HROPT) studies we observed that after subjects sat upright between series of supine and upright test swallows, they frequently had a transient lower oesophageal sphincter relaxation (TLOSR). When achalasia patients were studied in the same protocol, they exhibited a similar HROPT event leading to the hypothesis that achalasics had incomplete TLOSRs. We reviewed clinical HROPT studies of 94 consecutive non-achalasics and 25 achalasics. Studies were analyzed for a TLOSR like event during the study and, when observed, that TLOSR-like event was characterized for the degree and duration of distal oesophageal shortening, the degree of LOS relaxation, associated crural diaphragm (CD) inhibition, oesophageal pressurization and upper oesophageal sphincter (UOS) relaxation. About 64/94 (68%) non-achalasics and 15/24 (63%) of achalasics had a pressure topography event after the posture change characterized by a prolonged period of distal oesophageal shortening and/or LOS relaxation. Events among the non achalasics and achalasics were similar in terms of magnitude and duration of shortening and all were associated with CD inhibition. Similar proportions had associated non-deglutitive UOS relaxations. The only consistent differences were the absence of associated LOS relaxation and the absence of HROPT evidence of reflux among the achalasics leading us to conclude that their events were incomplete TLOSRs. Achalasic patients exhibit a selective defect in the TLOSR response suggesting preservation of all sensory, central and efferent aspects of the requisite neural substrate with the notable exception of LOS relaxation, a function of inhibitory (nitrergic) myenteric plexus neurons. PMID- 19552631 TI - Irritable bowel syndrome is strongly associated with generalized anxiety disorder: a community study. AB - BACKGROUND: No previous study has examined the comorbidity of Irritable Bowel Syndrome (IBS) and Generalized Anxiety Disorder (GAD) in a general population using standardized diagnostic methods. AIM: To examine the prevalence, comorbidity and risk correlates of IBS and GAD in a general population. METHODS: A random community-based telephone survey was conducted. The questionnaire covered symptoms of IBS, GAD, core depressive symptoms, help-seeking behaviour and functional impairment on the Sheehan Disability Scale. RESULTS: A total of 2005 participants completed the interview. The current prevalence of IBS was 5.4% and the 12-month prevalence of GAD was 4%. GAD was five times more common among IBS respondents than non-IBS respondents (OR: 5.84, P < 0.001), whereas IBS was 4.7 times more common among GAD respondents than among non-GAD respondents (OR: 6.32, P < 0.001). Core depressive symptoms (OR: 6.25, P < 0.01) and education level (OR: 5.918, P = 0.021) were risk correlates of GAD among IBS respondents. Comorbid respondents were more impaired than respondents having either disorder alone, but were not more likely to seek professional help than IBS-only respondents. CONCLUSION: Irritable Bowel Syndrome and GAD comorbidity was common and added to impairment in the community. The strong association between psychiatric morbidity and IBS observed in referral centres was not a consequence of increased help-seeking behaviour. PMID- 19552632 TI - Efficacy of methotrexate in Crohn's disease and ulcerative colitis patients unresponsive or intolerant to azathioprine /mercaptopurine. AB - BACKGROUND: Despite the wide use of azathioprine/mercaptopurine (AZA/MP) therapy in the management of both Crohn's disease (CD) and ulcerative colitis (UC), approximately 20% of patients cannot tolerate the drugs and 30% do not respond. AIM: To examine the efficacy and safety profile of methotrexate (MTX) in patients with CD or UC who are either intolerant or non-responsive to AZA/MP. METHODS: A total of 131 patients with IBD treated with MTX were identified. Retrospective data were obtained by case note review. Clinical response (defined as steroid withdrawal, normalization of previously raised CRP or physician's clinical assessment of improvement) was assessed at 6 months. RESULTS: Clinical response in Crohn's disease occurred in 18 of 29 patients (62%) refractory to AZA/MP and 42 of 70 patients (60%) intolerant to AZA/MP, with no difference between the groups (P = 1.0). In UC, clinical response was seen in 7 of 9 (78%) patients refractory to AZA/MP and 15 of 23 (65%) intolerant to thiopurines. MTX was well tolerated in a majority of individuals. CONCLUSIONS: Methotrexate appears effective in both CD and UC patients who fail to respond to or are intolerant to AZA/MP therapy. PMID- 19552633 TI - Prognostic impact of calf muscle near-infrared spectroscopy in patients with a first episode of deep vein thrombosis. AB - BACKGROUND: The risk factors that affect the development of post-thrombotic syndrome (PTS) are not fully recognized, and it is difficult to reliably predict which patients are likely to develop PTS in acute phase of deep vein thrombosis (DVT). AIMS: To investigate changes in calf muscle deoxygenated hemoglobin (HHb) levels after DVT, and to determine the indicative parameters reflecting the progression of PTS. METHODS: Seventy-six consecutive patients with a first episode of unilateral DVT were prospectively enrolled. Clinical manifestations were categorized according to the CEAP (Clinical, Etiologic, Anatomical, and Pathophysiologic) classification, and the patients were divided into no-PTS (C(0 3)E(s),A(s,d,p),P(r,o)) and PTS (C(4-6)E(s),A(s,d,p),P(r,o)) groups. Near infrared spectroscopy (NIRS) was used to measure calf muscle HHb levels at 6 months after diagnosis of DVT. The calf venous blood filling index (HHbFI) was calculated on standing, and the venous ejection index and the venous retention index (HHbRI) were then obtained after exercise. All patients were followed up for more than 24 months after the diagnosis of DVT. RESULTS: Of 76 patients evaluated, 20 (26.3%) had PTS. The NIRS-derived HHbFI and HHbRI were significantly increased in patients who developed PTS in comparison with those who did not (P = 0.04 and P = 0.0001, respectively). HHbRI was significantly increased in patients with iliofemoral DVT in comparison with patients with calf DVT (P = 0.041). An optimal cut-off point of 2.9 for HHbRI showed the strongest ability to predict the development of PTS, with a sensitivity of 100% and a specificity of 82.1%. CONCLUSIONS: HHbRI as measured by NIRS is significantly increased in patients with iliofemoral DVT as compared with those with calf DVT. Furthermore, HHbRI > 2.9 is a strong predictor of the development of PTS at 6 months. PMID- 19552634 TI - Ticagrelor binds to human P2Y(12) independently from ADP but antagonizes ADP induced receptor signaling and platelet aggregation. AB - BACKGROUND: P2Y(12) plays an important role in regulating platelet aggregation and function. This receptor is the primary target of thienopyridine antiplatelet agents, the active metabolites of which bind irreversibly to the receptor, and of newer agents that can directly and reversibly modulate receptor activity. OBJECTIVE: To characterize the receptor biology of the first reversibly binding oral P2Y(12) antagonist, ticagrelor (AZD6140), a member of the new cyclopentyltriazolopyrimidine (CPTP) class currently in phase III development. METHODS: Ticagrelor displayed apparent non-competitive or insurmountable antagonism of ADP-induced aggregation in human washed platelets. This was investigated using competition binding against [(3)H]ADP, [(33)P]2MeS-ADP and the investigational CPTP compound [(125)I]AZ11931285 at recombinant human P2Y(12). Functional receptor inhibition studies were performed using a GTPgammaS-binding assay, and further binding studies were performed using membranes prepared from washed human platelets. RESULTS: Radioligand-binding studies demonstrated that ticagrelor binds potently and reversibly to human P2Y(12) with K(on) and K(off) of (1.1 +/- 0.2) x 10(-4) nm(-1) s(-1) and (8.7 +/- 1.4) x 10(-4) s(-1), respectively. Ticagrelor does not displace [(3)H]ADP from the receptor (K(i) > 10 mum) but binds competitively with [(33)P]2MeS-ADP (K(i) = 4.3 +/- 1.3 nm) and [(125)I]AZ11931285 (K(i) = 0.33 +/- 0.04 nm), and shows apparent non-competitive inhibition of ADP-induced signaling but competitive inhibition of 2MeS-ADP induced signaling. Binding studies on membranes prepared from human washed platelets demonstrated similar non-competitive binding for ADP and ticagrelor. CONCLUSIONS: These data indicate that P2Y(12) is targeted by ticagrelor via a mechanism that is non-competitive with ADP, suggesting the existence of an independent receptor-binding site for CPTPs. PMID- 19552635 TI - Further evidence of genetic heterogeneity segregating with hereditary gingival fibromatosis. AB - AIM: To clinically characterize and map the disease-associated locus in a five generation Chinese family with autosomal dominant early-onset hereditary gingival fibromatosis (HGF). MATERIAL AND METHODS: A complete oral examination was conducted. Genomic DNA samples were obtained from 14 individuals. Short tandem repeats markers, which encompass four previously known loci related to HGF, were genotyped. Two-point log of the odds (LOD) scores were calculated using MLINK program of the LINKAGE software, multipoint and non-parametric linkage (NPL) analysis were performed using the GENEHUNTER software. RESULTS: Clinical evaluation and histological examination of this family suggested typical features of HGF. The onset age was early in the generations, ranging between 1 and 2 years. None of the tested markers showed cosegregation among affected individuals. Genotyping data from four putative regions yielded significant negative two-point LOD scores (<-2.0) at theta=0. The maximum multipoint LOD scores and NPL analysis revealed exclusion of these loci as well. CONCLUSIONS: Exclusion of linkage in this family to any of the known HGF loci proved the existence of a novel locus for autosomal dominant HGF and showed that this rare disorder is far more heterogeneous than previously expected. PMID- 19552636 TI - Transmitochondrial embryonic stem cells containing pathogenic mtDNA mutations are compromised in neuronal differentiation. AB - OBJECTIVES: Defects of the mitochondrial genome (mtDNA) cause a series of rare, mainly neurological disorders. In addition, they have been implicated in more common forms of movement disorders, dementia and the ageing process. In order to try to model neuronal dysfunction associated with mitochondrial disease, we have attempted to establish a series of transmitochondrial mouse embryonic stem cells harbouring pathogenic mtDNA mutations. MATERIALS AND METHODS: Transmitochondrial embryonic stem cell cybrids were generated by fusion of cytoplasts carrying a variety of mtDNA mutations, into embryonic stem cells that had been pretreated with rhodamine 6G, to prevent transmission of endogenous mtDNA. Cybrids were differentiated into neurons and assessed for efficiency of differentiation and electrophysiological function. RESULTS: Neuronal differentiation could occur, as indicated by expression of neuronal markers. Differentiation was impaired in embryonic stem cells carrying mtDNA mutations that caused severe biochemical deficiency. Electrophysiological tests showed evidence of synaptic activity in differentiated neurons carrying non-pathogenic mtDNA mutations or in those that caused a mild defect of respiratory activity. Again, however, neurons carrying mtDNA mutations that resulted in severe biochemical deficiency had marked reduction in post-synaptic events. CONCLUSIONS: Differentiated neurons carrying severely pathogenic mtDNA defects can provide a useful model for understanding how such mutations can cause neuronal dysfunction. PMID- 19552637 TI - Melatonin provides neuroprotection by reducing oxidative stress and HSP70 expression during chronic cerebral hypoperfusion in ovariectomized rats. AB - Oxidative stress is believed to contribute to functional and histopathologic disturbances associated with chronic cerebral hypoperfusion (CCH) in rats. Melatonin has protective effects against cerebral ischemia/reperfusion injury. This effect has mainly been attributed to its antioxidant properties. In the present study, we evaluate the effects of melatonin on chronic cerebral hypoperfused rats and examined its possible influence on oxidative stress, superoxide dismutase (SOD) activity, reduced glutathione (GSH) levels, and heat shock protein (HSP) 70 induction. CCH was induced by permanent bilateral common carotid artery occlusion in ovariectomized female rats. Extensive neuronal loss in the hippocampus at day 14 following CCH was observed. The ischemic changes were preceded by increases in malondialdehyde (MDA) concentration and HSP70 induction as well as reductions in GSH and SOD. Melatonin treatment restored the levels of MDA, SOD, GSH, and HSP70 induction as compared to the ischemic group. Histopathologic analysis confirmed the protective effect of melatonin against CCH induced morphologic alterations. Taken together, our results document that melatonin provides neuroprotective effects in CCH by attenuating oxidative stress and stress protein expression in neurons. This suggests melatonin may be helpful for the treatment of vascular dementia and cerebrovascular insufficiency. PMID- 19552638 TI - Effect of anti-platelet factor-4/heparin antibody induction on early saphenous vein graft occlusion after coronary artery bypass surgery. AB - BACKGROUND: Antibodies to complexes of heparin and platelet factor 4 (PF4) are capable of causing heparin-induced thrombocytopenia (HIT). Recent evidence suggests that anti-PF4/heparin antibodies may be prothrombogenic even in the absence of thrombocytopenia and clinically-recognized HIT. OBJECTIVES: To determine if induction of anti-PF4/heparin antibodies is an independent risk factor for early saphenous vein graft (SVG) occlusion or adverse clinical outcome after coronary artery bypass graft (CABG) surgery. PATIENTS/METHODS: Anti PF4/heparin antibody titers were measured in 368 patients prior to and then 4 days, 6 weeks and 6 months after CABG surgery. Serotonin release assay (SRA) and antibody isotype analysis were also performed on 6-week samples. SVG patency was determined in 297 patients 6 months after surgery by multidetector computed tomography coronary angiography. RESULTS: Six weeks after surgery, 52% of patients were anti-PF4/heparin seropositive and 9% were SRA positive. Six months after surgery, neither the percentage of occluded SVG (19% vs. 20%, P = NS), the percentage of patients with an occluded SVG (33% vs. 33%, P = NS) nor the incidence of adverse clinical events (21% vs. 24%, P = NS) differed between seropositive and seronegative groups. Neither IgG isotype nor SRA positivity was additionally predictive of SVG occlusion or adverse clinical outcome. CONCLUSION: Induction of anti-PF4/heparin antibodies, even those capable of heparin-dependent platelet activation, is not independently associated with early SVG occlusion or adverse clinical outcomes after CABG surgery. PMID- 19552639 TI - Chronic venous insufficiency is associated with elevated level of circulating microparticles. AB - BACKGROUND: Chronic venous insufficiency (CVI) results when the veins in the legs no longer pump blood back to the heart effectively. Microparticles (MPs) are small membrane vesicles released by several circulating and vascular cells upon activation or apoptosis. OBJECTIVES: The purpose of this study was to assess the subpopulations of circulating endothelial (EMPs) and platelet microparticles (PMPs) in CVI, and to disclose their contribution in mediating dysfunction of human peripheral venules. PATIENTS AND METHODS: Human peripheral venules were explanted during leg surgery on patients with CVI and on control subjects (C); concurrently, blood samples were collected and circulating MPs isolated. The techniques used were: flow cytometry, fluorescence and electron microscopy, myograph technique and western-blotting technique. RESULTS: The results showed that compared with controls, patients with CVI had: (i) a marked elevation of circulating EMPs and PMPs; (ii) a structural modification of the venous wall consisting of activation of endothelial and smooth muscle cells, an abundance of intermediary filaments and synthesis of hyperplasic-multilayered basal lamina; (iii) a significantly altered reactivity of the venous wall, closely associated with EMPs and PMPs adherence; (iv) altered contractile response to noradrenaline, acetylcholine, 5-hydroxytryptamine and KCl, and an impeded relaxation in response to sodium nitroprusside; and (iv) a substantially increased protein expression of tissue factor (TF) and of P-Selectin both in the venular vascular wall and on the surface of EMPs and PMPs. CONCLUSIONS: The findings indicate that CVI is accompanied by an enhanced release of EMPs and PMPs that contribute to altered dysfunctional response of the venous wall. PMID- 19552641 TI - Neonatal sucking behavior: comparison of perioral movement during breast-feeding and bottle feeding. AB - BACKGROUND: The aim of the present study was to compare orofacial motion and mouth angle during breast-feeding and bottle feeding in normal infants. METHODS: Thirteen normal infants were enrolled in the study, and feeding sessions were recorded using a digital video camera at 4 or 5 days following birth. The analysis was successfully performed for 12 infants during both breast-feeding and bottle feeding. Markers were placed at the lateral angle of the eye, the tip of the jaw, and the throat region. Jaw and throat region movements were calculated using the direct linear transformation method. The angle of the mouth was also measured. RESULTS: Throat region movement was significantly larger than jaw movement in both breast-feeding (eye-throat region changes: 1.51 +/- 0.37 mm, eye jaw changes: 0.91 +/- 0.31 mm, P < 0.001) and bottle feeding (eye-throat region changes: 2.91 +/- 0.46 mm, eye-jaw changes: 1.77 +/- 0.33 mm, P < 0.001). Jaw and throat region movements were significantly smaller during breast-feeding than those during bottle feeding (P < 0.001). The angle of the mouth during breast feeding is larger than that during bottle feeding (P < 0.001). There was a negative correlation between the angle of the mouth and jaw movement. CONCLUSIONS: Jaw and throat region movements during bottle feeding differ from those during breast-feeding. This partly results from the difference in the angle of the mouth during feeding. PMID- 19552640 TI - A low dose of Mycoplasma pneumoniae infection enhances an established allergic inflammation in mice: the role of the prostaglandin E2 pathway. AB - BACKGROUND: Over 40% of chronic stable asthma patients have evidence of respiratory Mycoplasma pneumoniae (Mp) infection as detected by PCR, but not by serology and culture, suggesting that a low-level Mp is involved in chronic asthma. However, the role of such a low-level Mp infection in the regulation of allergic inflammation remains unknown. OBJECTIVE: To determine the impact of a low-level Mp infection in mice with established airway allergic inflammation on allergic responses such as eosinophilia and chemokine eotaxin-2, and the underlying mechanisms [i.e. the prostaglandin E(2) (PGE(2)) pathway] since PGE(2) inhalation before an allergen challenge suppressed the eosinophil infiltration in human airways. METHODS: BALB/c mouse models of ovalbumin (OVA)-induced allergic asthma with an ensuing low- or high-dose Mp were used to assess IL-4 expression, bronchoalveolar lavage (BAL) eosinophil, eotaxin-2 and PGE(2) levels, and lung mRNA levels of microsomal prostaglandin E synthase-1 (mPGES-1). Primary alveolar macrophages (pAMs) from naive BALB/c mice were cultured to determine whether Mp induced PGE(2) or exogenous PGE(2) down-regulates IL-4/IL-13-induced eotaxin-2. RESULTS: Low-dose Mp in allergic mice significantly enhanced IL-4 and eotaxin-2, and moderately promoted lung eosinophilia, whereas high-dose Mp significantly reduced lung eosinophilia and tended to decrease IL-4 and eotaxin-2. Moreover, in both OVA-naive and allergic mice, lung mPGES-1 mRNA and BAL PGE(2) levels were elevated in mice infected with high-dose, but not low-dose Mp. In pAMs, IL-4/IL 13 significantly increased eotaxin-2, which was reduced by Mp infection accompanied by dose-dependent PGE(2) induction. Exogenous PGE(2) inhibited IL 4/IL-13-induced eotaxin-2 in a dose-dependent manner. CONCLUSIONS: This study highlights a novel concept on how different bacterial loads in the lung modify the established allergic airway inflammation and thus interact with an allergen to further induce Th2 responses. That is, unlike high-level Mp, low-level Mp fails to effectively induce PGE(2) to down-regulate allergic responses (e.g. eotaxin-2), thus maintaining or even worsening allergic inflammation in asthmatic airways. PMID- 19552642 TI - A comprehensive approach to risk assessment and surveillance guiding public health interventions. AB - Tools are required to identify 'multi-endemic' population segments - in order to benefit the most vulnerable people and to make public health interventions cost effective. The article suggests a comprehensive risk assessment and surveillance system approach based on a combination of epidemiological, environmental and social determinants. Such a cross-disciplinary approach will combine the advantages of forecasting upcoming disease 'hot spots' with provision of evidence for long-term planning under more stable conditions, and it may in principle apply to any combination of public health problems depending on the local context. PMID- 19552643 TI - Severe malaria and concomitant bacteraemia in children admitted to a rural Mozambican hospital. AB - OBJECTIVES: To describe the prevalence, aetiology and prognostic implications of coexisting invasive bacterial disease in children admitted with severe malaria in a rural Mozambican Hospital. METHODS: Retrospective study of data systematically collected from June 2003 to May 2007 in a rural Mozambican hospital, from all children younger than 5 years admitted with severe malaria. RESULTS: Seven thousand and forty-three children were admitted with a diagnosis of malaria. 25.2% fulfilled the criteria for severe malaria. 5.4% of the children with severe malaria and valid blood culture results had a concomitant bacteraemia. Case fatality rates of severe malaria cases rose steeply when bacteraemia was also present (from 4.0% to 22.0%, P < 0.0001), and bacteraemia was an independent risk factor for death among severe malaria patients (adjusted OR 6.2, 95% CI 2.8-13.7, P = 0.0001). Streptococcus pneumoniae, Gram-negative bacteria, Staphilococcus aureus and non-typhoid Salmonella (NTS) were the most frequently isolated microorganisms among severe malaria cases. Their frequency and associated case fatality rates (CFR) varied according to age and to syndromic presentation. Streptococcus pneumoniae had a relatively low CFR, but was consistently associated with severe malaria syndromes, or anaemia severity groups. No clear cut relationship between malarial anaemia and NTS bacteraemia was found. CONCLUSIONS: The coexistence of malaria and invasive bacterial infections is a frequent and life-threatening condition in many endemic African settings. In Mozambique, S. pneumoniae is the leading pathogen in this interaction, possibly as a consequence of the high HIV prevalence in the area. Measures directed at reducing the burden of both those infections are urgently needed to reduce child mortality in Africa. PMID- 19552644 TI - Three estimates of the association between linear growth failure and cognitive ability. AB - OBJECTIVES: To compare three estimators of association between growth stunting as measured by height-for-age Z-score and cognitive ability in children, and to examine the extent statistical adjustment for covariates is useful for removing confounding due to socio-economic status. METHODS: Three estimators, namely random-effects, within- and between-cluster estimators, for panel data were used to estimate the association in a survey of 1105 pairs of siblings who were assessed for anthropometry and cognition. Furthermore, a 'combined' model was formulated to simultaneously provide the within- and between-cluster estimates. RESULTS: Random-effects and between-cluster estimators showed strong association between linear growth and cognitive ability, even after adjustment for a range of socio-economic variables. In contrast, the within-cluster estimator showed a much more modest association: For every increase of one Z-score in linear growth, cognitive ability increased by about 0.08 standard deviation (P < 0.001). The combined model verified that the between-cluster estimate was significantly larger than the within-cluster estimate (P = 0.004). CONCLUSION: Residual confounding by socio-economic situations may explain a substantial proportion of the observed association between linear growth and cognition in studies that attempt to control the confounding by means of multivariable regression analysis. The within-cluster estimator provides more convincing and modest results about the strength of association. PMID- 19552645 TI - Risk factors for in-hospital mortality of visceral leishmaniasis patients in eastern Uganda. AB - OBJECTIVE: To identify risk factors for in-hospital mortality in patients treated for visceral leishmaniasis (VL) in Uganda. METHODS: Retrospective analysis of VL patients' clinical data collected for project monitoring by Medecins Sans Frontieres in Amudat, eastern Uganda. RESULTS: Between 2000 and 2005, of 3483 clinically suspect patients, 53% were confirmed with primary VL. Sixty-two per cent were children <16 years of age with a male/female ratio of 2.2. The overall case-fatality rate during pentavalent antimonial (n = 1641) or conventional amphotericin B treatment (n = 217) was 3.7%. There was no difference in the case fatality rate between treatment groups (P > 0.20). The main risk factors for in hospital death identified by a multivariate analysis were age <6 years and >15 years, concomitant tuberculosis or hepatopathy, and drug-related adverse events. The case-fatality rate among patients >45 years of age was strikingly high (29.0%). CONCLUSION: Subgroups of VL patients at higher risk of death during treatment with drugs currently available in Uganda were identified. Less toxic drugs should be evaluated and used in these patients. PMID- 19552646 TI - Feasibility, acceptability and cost of home-based HIV testing in rural Kenya. AB - OBJECTIVE: To demonstrate the feasibility, acceptability and cost of home-based HIV testing and to examine the applicability of the model to high HIV prevalence settings. METHODS: Quantitative, qualitative and cost data were collected during a home-based HIV testing program in a high-prevalence rural area of Kenya; data on age, gender and marital status along with HIV test results were collected. This was complemented with qualitative research including key informant interviews with counselors and program managers to highlight experiences and challenges. Direct costs of the interventions were estimated through the review of budgets and monthly expenditure sheets. RESULTS: Of 3180 15-49-year olds exposed to a community awareness campaign, 2033 (63.9%) agreed to be visited by counselors, of whom 1984 (97.6%) agreed to be tested and receive the results. Adult HIV prevalence was 8.2% and married women were 4.8 times more likely to be HIV-positive than those never married. Counselors reported feeling welcomed and noted the enthusiasm of the community towards testing. The total cost of the exercise was $17,569. The program cost was $2.60 for each of the 6750 community members, $5.88 for each person tested, and $84 per positive case detected. CONCLUSION: This study suggests that home-based HIV testing is feasible with high uptake, and has the potential to substantially expand access to HIV testing services. There is a strong economic case for the extension of such a screening program to other communities. PMID- 19552648 TI - Exo70-mediated recruitment of nucleoporin Nup62 at the leading edge of migrating cells is required for cell migration. AB - Nucleoporin Nup62 localizes at the central channel of the nuclear pore complex and is essential for nucleocytoplasmic transport. Through its FG-repeat domain, Nup62 regulates nuclear pore permeability and binds nuclear transport receptors. Here, we report that Nup62 interacts directly with Exo70 and colocalizes with Exo70 at the leading edge of migrating cells. Nup62 binds the N-terminal domain of Exo70 through its coiled-coil domain but not through its FG-repeat domain. Selective inhibition of leading edge Nup62 using RNA interference significantly reduces cell migration. Furthermore, Exo70 recruits Nup62 at the plasma membrane and at filopodia. Removal of the Exo70-binding domain of Nup62 prevents leading edge localization of Nup62. Analogous to Exo70, Nup62 cycles between the plasma membrane and the perinuclear recycling compartment. Altogether, we propose that Nup62 not solely regulates access to the cell nucleus, but additionally functions in conjunction with Exo70, a key regulator of exocytosis and actin dynamics, at the leading edge of migrating cells. PMID- 19552649 TI - Estimating stochastic elasticities directly from longitudinal data. AB - The elasticities of long-run population growth rate with respect to vital rates are useful in studying selection on vital rates, and in evaluating management policy that aims to control vital rates. In temporally varying environments, elasticity is often calculated from simulations that assume a probability distribution for the environmental states. Here we develop a method to estimate elasticities directly from demographic data. Using a time-series of demographic matrices and age-structure we construct a consistent statistical estimator of elasticity that converges to the correct limiting value as the sample length increases. We also construct confidence intervals for elasticities from temporal data and suggest tools for testing hypotheses about the strength of selection. We use data on a natural population to show that our method can indeed accurately estimate elasticities using relatively short time series. PMID- 19552650 TI - Photochemical decomposition of dipyridamole in aqueous solution and the utilization of citrate-theophylline-adenosine-dipyridamole anticoagulant for monitoring of heparin. PMID- 19552647 TI - Messenger RNA export from the nucleus: a series of molecular wardrobe changes. AB - The advent of the nucleus during the evolutionary development of the eukaryotic cell necessitated the development of a transport system to convey messenger RNA (mRNA) from the site of transcription in the nucleus to ribosomes in the cytoplasm. In this review, we highlight components of each step in mRNA biogenesis, from transcription to processing, that are coupled with mRNA export from the nucleus. We also review the mechanism by which proteins from one step in the mRNA assembly line are replaced by those required for the next. These 'molecular wardrobe changes' appear to be key steps in facilitating the rapid and efficient nuclear export of mRNA transcripts. PMID- 19552651 TI - Drug resistant ADLTE and recurrent partial status epilepticus with dysphasic features in a family with a novel LGI1mutation: electroclinical, genetic, and EEG/fMRI findings. AB - PURPOSE: We characterized a family with autosomal dominant lateral temporal epilepsy (ADLTE) whose proband presented uncommon electroclinical findings such as drug-resistant seizures and recurrent episodes of status epilepticus with dysphasic features. METHODS: The electroclinical characteristics and LGI1 genotype were defined in the family. In the proband, the ictal pattern was documented during video-EEG monitoring and epileptic activity was mapped by EEG/fMRI. RESULTS: The affected members who were studied had drug-resistant seizures. In the proband, seizures with predominant dysphasic features often occurred as partial status epilepticus. The video-EEG-documented ictal activity and fMRI activation clearly indicated the elective involvement of the left posterior lateral temporal cortex. Sequencing of LGI1 exons revealed a heterozygous c.367G>A mutation in exon 4, resulting in a Glu123Lys substitution in the protein sequence. CONCLUSIONS: The uncommon clinical pattern (high seizure frequency, drug-resistance) highlights the variability of the ADLTE phenotype and extends our knowledge of the clinical spectrum associated with LGI1 mutations. PMID- 19552652 TI - Structures involved at the time of temporal lobe spikes revealed by interindividual group analysis of EEG/fMRI data. AB - PURPOSE: We measured metabolic changes associated with temporal lobe (TL) spikes using combined electroencephalography (EEG) and functional magnetic resonance imaging (fMRI). We selected 18 patients with temporal lobe epilepsy (TLE) who underwent a 2-h simultaneous EEG-fMRI and had unilateral or bilateral independent TL spikes for interindividual group analysis, in order to identify consistent blood oxygenation level dependent (BOLD) responses to TL spikes. METHODS: EEG was postprocessed and spikes were visually identified. fMRI data were preprocessed with motion correction, spatial smoothing, and removal of low frequency drifts. Spike timings were used as events for fMRI statistical analysis. Four hemodynamic response functions were used to account for variability in the BOLD response. RESULTS: Group analysis revealed common areas of BOLD activations and deactivations. The hemodynamic response function (HRF) peaking 3 s after the spike showed activation involving ipsilaterally the mesial temporal structures (presumably the hippocampus), putamen/globus pallidus, inferior insula, and superior temporal gyrus. The HRF peaking at 5 s showed activations involving ipsi and contralaterally the superior temporal gyrus and inferior insula. Both HRFs showed bilateral posterior cingulate deactivations. DISCUSSION: We disclosed involvement of a network of activated areas during unilateral TL spikes, including ipsilateral mesial temporal structures, basal ganglia, and bilateral neocortical temporal regions. Despite the low temporal resolution of fMRI we demonstrated that contralateral temporal involvement occurred later than ipsilateral activation. This contralateral change took place in the absence of visible EEG changes. The posterior cingulate deactivation may reflect the interconnections between this region and other limbic structures. It may also partially correspond to a suspension of the default mode network, as previously described for TL spikes. PMID- 19552653 TI - Stiripentol open study in Japanese patients with Dravet syndrome. AB - PURPOSE: To survey the treatment situation of Dravet syndrome in Japan and to compare this result with effectiveness of stiripentol (STP) add-on therapy in an open-label multicenter study. METHODS: Medical records of patients with Dravet syndrome who visited the study institutions during 2006 were surveyed to examine the effect of antiepileptic drugs (AEDs) on clonic or tonic-clonic seizures (GTCS). Patients older than 1 year of age treated with at least one conventional AED and more than four GTCS per month were invited to participate in the STP study. Seizure status and adverse effects during the first 4 weeks of STP (50 or 1,000 mg/day) add-on therapy (early period) and during long-term treatment were compared with baseline. RESULTS: Only 15% of the treatment trials with 15 conventional AEDs in 112 patients succeeded in reducing seizures by more than 50%. With STP, GTCS were reduced more than 50% in 14 of 23 patients (61%), including 2 who became seizure-free, in the early period. Moreover, duration of seizures was shortened in 10 patients and status epilepticus decreased in 6. These effects continued in the long-term although to a lesser degree. Adverse effects (loss of appetite, sleep disturbance, ataxia, hyperactivity/irritability) disappeared after dose modification in most cases. STP was effective at a lower than initial dose in five patients. Some patients benefited from STP added on clobazam despite mutation in CYP2C19. CONCLUSION: Our data suggest that an early introduction of STP into Japan will result in substantial patient benefit. PMID- 19552654 TI - Seizure frequency and sex steroids in women with partial epilepsy on antiepileptic therapy. AB - PURPOSE: Neuroactive sex steroids influence neuron excitability, which is enhanced by estradiol (E2) and decreased by progesterone (Pg). In epilepsy, the production, metabolism, biologic availability, and activity of sex hormones may be affected by seizures themselves or by antiepileptic drugs (AEDs). This cross sectional observational study was aimed at evaluating the relationships between sex steroids, seizure frequency, and other clinical parameters in women with partial epilepsy (PE) on AED treatments. METHODS: Serum E2, Pg, sex hormone binding globulin (SHBG) levels, free E2 (fE2), and E2/Pg ratios were determined during the follicular and luteal phases in 72 adult women with PE, and in 30 healthy controls. Hormonal data were correlated with seizure frequency, age, body weight, body mass index (BMI), disease onset and duration, and AED therapies. RESULTS: In patients, E2, fE2, and Pg levels were lower in both ovarian phases, whereas those of SHBG were higher than in controls. No significant changes in hormone levels and in prevalence of anovulatory cycles were observed between patients grouped according to their seizure frequency. However, when compared with those in healthy controls, luteal fE2 and Pg levels were chiefly impaired in women with more frequent seizures, mostly undergoing AED polytherapies, but not in those with absent or rarer seizures. CONCLUSIONS: The actual changes in sex steroid levels and E2/Pg ratios did not explain an increased seizure frequency in adult women with AED-treated PE, but patients with more severe disease showed more relevant changes in their sex hormone profile and impaired Pg levels during the luteal phase. PMID- 19552657 TI - A model for community mental health services in South Africa. AB - OBJECTIVES: To provide a model to estimate human resource needs for community based mental health services in South Africa. METHODS: A situation analysis was conducted of current community-based mental health service provision in South Africa, which comprise outpatient and emergency services, residential care and day care. Service utilisation rates and staffing needs were estimated for two levels of service coverage, using data from the situation analysis, local epidemiological studies and consultation with key stakeholders. RESULTS: For a population of 100,000 people, 7.3-23.8 full-time equivalent staff would be required to provide services in outpatient services, 14.9-41.6 in day care and 11.5-23.0 in residential care at minimum and full coverage levels respectively. The model can facilitate rational planning by requiring transparency and accountability in the assumptions used. This method can be adapted to a range of countries, by entering relevant country data. CONCLUSIONS: The model fills a gap, particularly in low- and middle-income countries, where community-based mental health services are sparse, and decisions regarding allocations to them are hampered by a lack of good quality data. The results of the model are limited by the quality of data and the assumptions upon which the modelling are based. PMID- 19552658 TI - Pentamidine as secondary prophylaxis for visceral leishmaniasis in the immunocompromised host: report of four cases. AB - We report a retrospective and descriptive study of four immunocompromised patients (three with HIV-1 and one with idiopathic CD4+-lymphopenia) with relapsing visceral leishmaniasis seen at the Hospital for Tropical Diseases, London, in whom pentamidine was used as secondary prophylaxis to prevent relapse. Patients experienced between one and four relapses before commencing prophylaxis with subsequent relapse-free periods ranging from 5 to 98 months. Based on these observational data, we recommend large trials to investigate the efficacy of pentamidine over other agents in preventing relapse of VL in the immunocompromised patient. PMID- 19552656 TI - Distributed source modeling of language with magnetoencephalography: application to patients with intractable epilepsy. AB - PURPOSE: To examine distributed patterns of language processing in healthy controls and patients with epilepsy using magnetoencephalography (MEG), and to evaluate the concordance between laterality of distributed MEG sources and language laterality as determined by the intracarotid amobarbital procedure (IAP). METHODS: MEG was performed in 10 healthy controls using an anatomically constrained, noise-normalized distributed source solution (dynamic statistical parametric map, dSPM). Distributed source modeling of language was then applied to eight patients with intractable epilepsy. Average source strengths within temporoparietal and frontal lobe regions of interest (ROIs) were calculated, and the laterality of activity within ROIs during discrete time windows was compared to results from the IAP. RESULTS: In healthy controls, dSPM revealed activity in visual cortex bilaterally from approximately 80 to 120 ms in response to novel words and sensory control stimuli (i.e., false fonts). Activity then spread to fusiform cortex approximately 160-200 ms, and was dominated by left hemisphere activity in response to novel words. From approximately 240 to 450 ms, novel words produced activity that was left-lateralized in frontal and temporal lobe regions, including anterior and inferior temporal, temporal pole, and pars opercularis, as well as bilaterally in posterior superior temporal cortex. Analysis of patient data with dSPM demonstrated that from 350 to 450 ms, laterality of temporoparietal sources agreed with the IAP 75% of the time, whereas laterality of frontal MEG sources agreed with the IAP in all eight patients. DISCUSSION: Our results reveal that dSPM can unveil the timing and spatial extent of language processes in patients with epilepsy and may enhance knowledge of language lateralization and localization for use in preoperative planning. PMID- 19552659 TI - Nutritional status of Malawian adults on antiretroviral therapy 1 year after supplementary feeding in the first 3 months of therapy. AB - OBJECTIVE: To test the hypothesis that individuals on antiretroviral therapy (ART) for 3 months with a greater body mass index (BMI) as a result of supplementary feeding with ready-to-use fortified spread would maintain a higher BMI 9 months after the feeding ended. METHODS: Two cohorts of wasted adults with AIDS, after 12 months of ART and 3 months of supplementary feeding with either ready-to-use fortified spread, an energy dense lipid paste; or corn/soy blended flour, were assessed for clinical and anthropometric status, quality of life, and ART adherence after 3 and 9 months. RESULTS: 336 ART patients participated: 162 who had received ready-to-use fortified spread and 174 who had received corn/soy blended flour. 9 months after stopping food supplements, both groups had a similar BMI, fat-free body mass, hospitalization rate and mortality. Binary logistic regression modelling showed that lower BMI, lower CD4 count, and older age at baseline were associated with a higher risk of death (odds ratio for BMI = 0.63, 95% CI 0.47-0.79). Adherence to the ART regimen and quality of life were similar in both cohorts. CONCLUSION: While supplementary feeding with ready-to use fortified spread can ameliorate the BMI, an established risk factor for mortality, this effect is sustained only during the time of the intervention. Supplementary feeding of wasted patients for longer than 3 months should be investigated. PMID- 19552655 TI - Pursuing paradoxical proconvulsant prophylaxis for epileptogenesis. AB - There are essentially two potential treatment options for any acquired disorder: symptomatic or prophylactic. For acquired epilepsies that follow a variety of different brain insults, there remains a complete lack of prophylactic treatment options, whereas at the same time these epilepsies are notoriously resistant, once they have emerged, to symptomatic treatments with antiepileptic drugs. The development of prophylactic strategies is logistically challenging, both for basic researchers and clinicians. Nevertheless, cannabinoid-targeting drugs provide a very interesting example of a system within the central nervous system (CNS) that can have very different acute and long-term effects on hyperexcitability and seizures. In this review, we outline research on cannabinoids suggesting that although cannabinoid antagonists are acutely proconvulsant, they may have beneficial effects on long-term hyperexcitability following brain insults of multiple etiologies, making them promising candidates for further investigation as prophylactics against acquired epilepsy. We then discuss some of the implications of this finding on future attempts at prophylactic treatments, specifically, the very short window within which prevention may be possible, the possibility that traditional anticonvulsants may interfere with prophylactic strategies, and the importance of moving beyond anticonvulsants-even to proconvulsants-to find the ideal preventative strategy for acquired epilepsy. PMID- 19552660 TI - Outbreak of leptospirosis in New Caledonia: diagnosis issues and burden of disease. AB - A leptospirosis epidemic affected New Caledonia during the first semester of 2008. A total of 135 cases were diagnosed with a relatively low fatality rate of 3.7%. Heavy rainfalls, related to La Nina, favoured this epidemic. The PCR, routinely used, confirmed 54% of the cases, and the microagglutination test 56%. Epidemiological and economical data on this epidemic are presented and discussed. PMID- 19552662 TI - Prolonged persistence of residual Wuchereria bancrofti infection after cessation of diethylcarbamazine-fortified salt programme. AB - A diethylcarbamazine (DEC)-fortified salt intervention programme was implemented between 1982 and 1986 in Karaikal district, Union territory of Pondicherry, south India, to control Culex transmitted bancroftian filariasis. The intervention reduced the microfilaria (Mf) rate from 4.49% to 0.08%. To eliminate the residual microfilaraemia, the health department detected and treated Mf carriers from 1987 to 2005 and mass-administered drugs in 2004 and 2005. Surveillance from 1987 to 2005 revealed persistent microfilaraemia in 0.03-0.42% of the population. In 2006, we conducted a more detailed Mf survey and a child antigenaemia (Ag) survey in 15 urban wards and 17 rural villages. These surveys showed an overall Mf rate of 0.46% in the high-risk urban areas and 0.18% in the rural areas; none of the sampled children was positive for Ag. All detected Mf carriers were >20 years old. The age of the youngest Mf carrier was 30 years in urban and 21 years in rural areas, which suggests that transmission was interrupted and there was no incidence of new Mf case after cessation of DEC salt programme. Eleven of 15 urban and 15 of 17 villages were totally free from microfilaraemia. Nevertheless, three of 15 surveyed urban localities and two of 17 villages showed >1% Mf rate. Thus, it seems that (i) post-intervention very low levels of microfilaraemia can continue as long as 20 years; (ii) 0.60-0.70% Mf rate is a safe level and at this level recrudescence of infection may not occur; (iii) there can be isolated localities with >1% Mf rate and their detection for further intervention measures could be challenging in larger control/elimination programmes and (iv) the residual infection mostly gets concentrated in the adult population, in underdeveloped urban areas and in historically highly endemic or large endemic rural areas. These groups and areas should be targeted with rigorous intervention measures such as mass drug administration to eliminate the residual infection. PMID- 19552661 TI - Diagnosis of antiretroviral therapy failure in Malawi: poor performance of clinical and immunological WHO criteria. AB - OBJECTIVES: In antiretroviral therapy (ART) scale-up programmes in sub-Saharan Africa viral load monitoring is not recommended. We wanted to study the impact of only using clinical and immunological monitoring on the diagnosis of virological ART failure under routine circumstances. METHODS: Clinicians in two urban ART clinics in Malawi used clinical and immunological monitoring to identify adult patients for switching to second-line ART. If patients met clinical and/or immunological failure criteria of WHO guidelines and had a viral load <400 copies/ml there was misclassification of virological ART failure. RESULTS: Between January 2006 and July 2007, we identified 155 patients with WHO criteria for immunological and/or clinical failure. Virological ART failure had been misclassified in 66 (43%) patients. Misclassification was significantly higher in patients meeting clinical failure criteria (57%) than in those with immunological criteria (30%). On multivariate analysis, misclassification was associated with being on ART <2 years [OR = 7.42 (2.63, 20.95)] and CD4 > 200 cells/microl [OR = 5.03 (2.05, 12.34)]. Active tuberculosis and Kaposi's sarcoma were the most common conditions causing misclassification of virological ART failure. CONCLUSION: Misclassification of virological ART failure occurs frequently using WHO clinical and immunological criteria of ART failure for poor settings. A viral load test confirming virological ART failure is therefore advised to avoid unnecessary switching to second-line regimens. PMID- 19552664 TI - Ribavirin dose reduction raises relapse rate dose-dependently in genotype 1 patients with hepatitis C responding to pegylated interferon alpha-2b plus ribavirin. AB - The impact of ribavirin exposure on virologic relapse remains controversial in combination therapy with pegylated interferon (Peg-IFN) and ribavirin for patients with chronic hepatitis C (CH-C) genotype 1. The present study was conducted to investigate this. Nine hundred and eighty-four patients with CH-C genotype 1 were enrolled. The drug exposure of each medication was calculated by averaging the dose actually taken. For the 472 patients who were HCV RNA negative at week 24 and week 48, multivariate logistic regression analysis showed that the degree of fibrosis (P = 0.002), the timing of HCV RNA negativiation (P < 0.001) and the mean doses of ribavirin (P < 0.001) were significantly associated with relapse, but those of Peg-IFN were not. Stepwise reduction of the ribavirin dose was associated with a stepwise increase in relapse rate from 11% to 60%. For patients with complete early virologic response (c-EVR) defined as HCV RNA negativity at week 12, only 4% relapse was found in patients given > or = 12 mg/kg/day of ribavirin and ribavirin exposure affected the relapse even after treatment week 12, while Peg-IFN could be reduced to 0.6 microg/kg/week after week 12 without the increase of relapse rate. Ribavirin showed dose-dependent correlation with the relapse. Maintaining as high a ribavirin dose as possible (> or = 12 mg/kg/day) during the full treatment period can lead to suppression of the relapse in HCV genotype 1 patients responding to Peg-IFN alpha-2b plus ribavirin, especially in c-EVR patients. PMID- 19552663 TI - Pegylated interferon alpha-2b (Peg-IFN alpha-2b) affects early virologic response dose-dependently in patients with chronic hepatitis C genotype 1 during treatment with Peg-IFN alpha-2b plus ribavirin. AB - Chronic hepatitis C (CH-C) genotype 1 patients who achieved early virologic response have a high probability of sustained virologic response (SVR) following pegylated interferon (Peg-IFN) plus ribavirin therapy. This study was conducted to evaluate how reducing drug doses affects complete early virologic response (c EVR) defined as hepatitis C virus (HCV) RNA negativity at week 12. Nine hundred eighty-four patients with CH-C genotype 1 were enrolled. Drug doses were evaluated independently on a body weight base from doses actually taken. From multivariate analysis, the mean dose of Peg-IFN alpha-2b during the first 12 weeks was the independent factor for c-EVR (P = 0.02), not ribavirin. The c-EVR rate was 55% in patients receiving > or = 1.2 microg/kg/week of Peg-IFN, and declined to 38% at 0.9-1.2 microg/kg/week, and 22% in patients given <0.9 microg/kg/week (P < 0.0001). Even with stratified analysis according to ribavirin dose, the dose-dependent effect of Peg-IFN on c-EVR was observed, and similar c EVR rates were obtained if the dose categories of Peg-IFN were the same. Furthermore, the mean dose of Peg-IFN during the first 12 weeks affected HCV RNA negativity at week 24 (P < 0.0001) and SVR (P < 0.0001) in a dose-dependent manner. Our results suggest that Peg-IFN was dose-dependently correlated with c EVR, independently of ribavirin dose. Thus, maintaining the Peg-IFN dose as high as possible during the first 12 weeks can yield HCV RNA negativity and higher c EVR rates, leading to better SVR rates in patients with CH-C genotype 1. PMID- 19552665 TI - Sex-related adaptive responses of Populus cathayana to photoperiod transitions. AB - Populus cathayana Rehd., a dioecious tree species, occupies a wide range of habitats in southwest China. Both males and females were grown under two regimes of photoperiod, from mid-length to short-day photoperiod (SD shift), or to long day photoperiod (LD shift). SD shift triggered leaf senescence in both males and females by decreasing net photosynthesis rate (A), transpiration (E), and chlorophyll pigment (Chl), non-structural carbohydrate (NSC) and indoleacetic acid (IAA) contents, while increasing abscisic acid (ABA), malonaldehyde (MDA) and free proline (Pro) contents. The antioxidant enzyme (e.g. POD, CAT and SOD) activities and capability to maintain ultrastructural integrity also decreased under SD shift. Males exhibited faster leaf senescence than did females, as shown by greater decreases in A, E, Chl and IAA. However, males maintained a less senescent stage than did females, as indicated by higher values of A, Chl, NSC, IAA and antioxidant enzyme activities. Conversely, A, E, NSC and IAA contents and antioxidant enzyme activities were enhanced by lower O(2)(*-) in females, whereas reduced by higher O(2)(*-) in males under LD shift. Such sex-dependent responses of P. cathayana to photoperiod transitions showed that males and females possess different adaptabilities, which may relate to sex-specific leaf senescence speed under changing environments. PMID- 19552666 TI - Transfer of nitrogen from a tropical legume tree to an associated fodder grass via root exudation and common mycelial networks. AB - Symbiotic dinitrogen fixation by legume trees represents a substantial N input in agroforestry systems, which may benefit the associated crops. Applying (15)N labelling, we studied N transfer via common mycelial networks (CMN) and root exudation from the legume tree Gliricidia sepium to the associated fodder grass Dichantium aristatum. The plants were grown in greenhouse in shared pots in full interaction (treatment FI) or with their root systems separated with a fine mesh that allowed N transfer via CMN only (treatment MY). Tree root exudation was measured separately with hydroponics. Nitrogen transfer estimates were based on the isotopic signature of N (delta(15)N) transferred from the donor. We obtained a range for estimates by calculating transfer with delta(15)N of tree roots and exudates. Nitrogen transfer was 3.7-14.0 and 0.7-2.5% of grass total N in treatments FI and MY, respectively. Root delta(15)N gave the lower and exudate delta(15)N the higher estimates. Transfer in FI probably occurred mainly via root exudation. Transfer in MY correlated negatively with grass root N concentration, implying that it was driven by source-sink relationships between the plants. The range of transfer estimates, depending on source delta(15)N applied, indicates the need of understanding the transfer mechanisms as a basis for reliable estimates. PMID- 19552667 TI - The involvement of aquaglyceroporins in transport of boron in barley roots. AB - Boron (B) enters cells as the uncharged boric acid, a small neutral molecule with sufficient lipid solubility to cross cell membranes without the aid of transport proteins. The extent to which the observed uptake rates for B in plants can be explained by this simple physical process was examined by applying treatments expected to inhibit the membrane transporters most likely to be involved in B transport. These experiments established that at least 50% of B uptake could be facilitated by transporters. The B transport characteristics of two barley aquaglyceroporins, HvPIP1;3 and HvPIP1;4, were investigated using yeast complementation assays. Expression of both genes in yeast resulted in increased B sensitivity. Transport assays in yeast confirmed that HvPIP1;3 and HvPIP1;4 are both capable of transporting B. The physiological role of these HvPIP1 genes in B transport is uncertain since their expression was not responsive to B nutritional status, and they continued to be expressed under toxicity conditions. PMID- 19552668 TI - Bulk leaf delta(18)O and delta(13)C reflect the intensity of intraspecific competition for water in a semi-arid tussock grassland. AB - We investigated the extent to which plant water and nutrient status are affected by intraspecific competition intensity and microsite quality in a monodominant tussock grassland. Leaf gas exchange and stable isotope measurements were used to assess the water relations of Stipa tenacissima tussocks growing along a gradient of plant cover and soil depth in a semi-arid catchment of Southeast Spain. Stomatal conductance and photosynthetic rate decreased with increasing intensity of competition during the wet growing season, leading to foliar delta(18)O and delta(13)C enrichment. A high potential for runoff interception by upslope neighbours exerted strong detrimental effects on the water and phosphorus status of downslope S. tenacissima tussocks. Foliar delta(15)N values became more enriched with increasing soil depth. Multiple stepwise regression showed that competition potential and/or rhizosphere soil depth accounted for large proportions of variance in foliar delta(13)C, delta(18)O and delta(15)N among target tussocks (57, 37 and 64%, respectively). The results presented here highlight the key role that spatial redistribution of resources (water and nutrients) by runoff plays in semi-arid ecosystems. It is concluded that combined measurement of delta(13)C, delta(18)O and nutrient concentrations in bulk leaf tissue can provide insight into the intensity of competitive interactions occurring in natural plant communities. PMID- 19552669 TI - The effect of the first application of verteporfin photodynamic therapy on lesion growth in choroidal neovascularisation and its potential impact on combination therapy. AB - PURPOSE: To evaluate the impact of lesion size on the observed growth of the choroidal neovascularisation (CNV) following the first application of photodynamic therapy (PDT). METHODS: A retrospective study of consecutive patients with subfoveal classic CNV or predominantly classic CNV undergoing first verteporfin photodynamic therapy (VPDT) according to treatment of age-related macular degeneration with photodynamic therapy (TAP) protocol between June 2005 and September 2005. Patients were classified into two groups according to baseline greatest linear dimension (GLD) at the time of VPDT monotherapy. Group 1 comprised patients with lesion GLD less then 2000 microm and Group 2, patients with lesion GLD 2000 microm or larger. Difference between the mean GLD at baseline and at three months post-treatment were investigated for clinical significance using the paired t-test. RESULTS: Group 1 (n = 16) showed a mean change in GLD from 1380.25 microm to 2031.25 microm, while Group 2 (n = 26) showed a mean change of 2909.26 microm to 3023.07 microm. Data of mean percentage change in GLD for Group 1 showed a 51.97 per cent increase in the lesion size [95% CI, 22.93 - 81.01%] in comparison to Group 2, which showed only 5.8 per cent increase in the lesion [95% CI, -9.65 - 21.61%]. Comparison of the mean percentage change in GLD between the two groups showed a statistically significant difference (p = 0.003). CONCLUSIONS: Our study demonstrates that following the first application of VPDT, smaller lesions enlarge disproportionately more than larger lesions. As we have entered into the era of using angiostatic agents in combination with VPDT, it may be important to evaluate this effect with regards to the timing of VPDT commencement. PMID- 19552670 TI - Feasibility of using tissue Doppler velocities in stress echo during upright bicycle exercise. AB - BACKGROUND AND AIM: There are several studies on myocardial tissue Doppler velocities during dobutamine stress, but few studies on the same parameters during bicycle exercise. We wanted to examine how sample sites affected velocities, beat-to-beat variability and segments eligible for analysis. METHODS: Twenty patients with normal coronary arteries were included in the study. Echocardiograms were obtained at rest and at peak exercise on the exercise bike and were analyzed off line by two independent raters. RESULTS AND CONCLUSIONS: Tissue velocities decreased from base to apex. Mean difference between the raters was-0.02 cm/sec at rest and 0.45 cm/sec during exercise. Peak-to-peak variability was lowest in the mitral ring during rest (10-14%) and increased at peak exercise (15-21%). The proportion of segments eligible for analysis was 84% in the septum and 67% in the anterior wall during exercise. Thus, in patients with normal coronary arteries, tissue Doppler velocities decrease from base toward the apex during bicycle exercise. Interrater agreement is reduced during exercise and beat to-beat variability is lowest in the mitral ring. PMID- 19552671 TI - Mitral regurgitation associated with mitral annular dilation in patients with lone atrial fibrillation: an echocardiographic study. AB - BACKGROUND: Whether and how lone atrial fibrillation (AF) is associated with functional mitral regurgitation (MR) remain unclear. METHOD: We studied 12 lone AF patients without left ventricular (LV) dysfunction and/or dilatation, who underwent mitral valve annuloplasty for functional mitral regurgitation (MR). Ten lone AF patients without MR served as controls. RESULTS: Lone AF Patients with MR had a greater mitral valve annular area and left atrial area than those without MR. There were no differences in LV volumes or LV ejection fraction. CONCLUSIONS: Therefore, we concluded that left atrial dilation and corresponding mitral annular dilation may cause MR in lone AF patients without LV dysfunction and/or dilatation. PMID- 19552672 TI - Inferior vena cava syndrome due to Echinococcus multilocularis. AB - Alveolar echinococcosis is an invasive, tumor-like zoonosis, accidentally transmitted to humans. We present a case of recurrent inferior vena cava (IVC) syndrome due to alveolar echinococcosis and strongly suspected on transthoracic echocardiographic examination. PMID- 19552673 TI - Arterial mechanics in ischemic versus nonischemic cardiomyopathy: clinical and diagnostic impact. AB - BACKGROUND: Differential diagnosis between ischemic (IDCM) and the nonischemic type (NIDCM) of cardiomyopathy constitutes a challenge in the daily medical practice. Carotid and aortic elastic properties deteriorate in patients with coronary artery disease. However, their predictive role in differentiating IDCM from NIDCM has not been addressed so far. AIM OF THE WORK: To examine carotid and aortic mechanical functions using conventional and Doppler tissue echocardiography in the distinction between IDCM and NIDCM in patients with clinically undetermined etiology. METHODS: 70 patients with dilatation and diffuse impairment of the left ventricular (LV) contraction were studied. All patients underwent carotid duplex for measuring intima-media (IMT) thickness, peak systolic velocity (PSV), and luminal diameters (LD). Aortic distensibility, strain, and aortic wall velocities (systolic (Sa), early diastolic (Ea), late diastolic (Aa) velocities, Sa(t), and Ea(t)) were measured. According to coronary angiographic results, patients were categorized into IDCM (n = 36) (age 57.9 +/- 9.2 years) and NIDCM groups (n = 34) (age 56.0 +/- 8.3 years); they were compared to 30 age- and sex-matched healthy individuals as a control group. RESULTS: The aortic pulsatile change, aortic strain, and distensibility were significantly reduced in both patient groups in comparison to control (P < 0.001). These parameters were much impaired in patients with IDCM compared with NIDCM (P < 0.001). IDCM have more deterioration of Sa, Ea, and Aa compared with NIDCM group (7.6 +/- 2.4 vs. 8.9 +/- 1.58, 7.5 +/- 2.8 vs. 10.6 +/- 1.5, 9.0 +/- 1.4 vs. 6.9 +/- 2.4 cm/sec; P < 0.001), respectively. In IDCM, the variables of aortic elastic properties were correlated only to age, while in NIDCM they were correlated to hemodynamics, LV volumes, wall thickness, and mass. Both carotid diameter and IMT were significantly increased in IDCM in comparison to NIDCM and control (P < 0.001). Carotid distensibility was significantly reduced in IDCM compared with NIDCM and control (P < 0.001). However, the carotid properties strongly correlated to risk factors in IDCM and to hemodynamics and LV function in NIDCM. Using ROC curve, a cutoff value < or =4.7 (cm(2)/dyne/10(3)) for aortic distensibility, value <8 cm/sec for Sa and IMT >0.8 mm predicted IDCM with 94.4%, 72.7%, and 97.2% sensitivity and 88.2%, 85.3%, and 97.1% specificity, respectively. CONCLUSION: Both carotid and aortic mechanical functions are more deteriorated in ischemic compared with nonischemic dilated cardiomyopathy. Different functional and structural mechanisms might be responsible for the deterioration of arterial elastic properties in each category. PMID- 19552674 TI - Dental care visits among dentate adults with diabetes and periodontitis. AB - OBJECTIVES: Two national surveys have shown that dentate adults with diabetes are less likely to visit a dentist than are those without diabetes; one survey showed this association only among women. We hypothesize that periodontal health among those with diabetes could explain this disparity. This report investigates the influence of periodontitis on the association between diabetes and dental care visits. It also tests whether disparities are limited to women. METHODS: Data from the 1999-2004 National Health and Nutrition Examination Survey were used. Covariates included age, sex, race/ethnicity, poverty status, education level, dental insurance, and periodontitis status. Weighted analyses were limited to dentate adults aged > or =25 years. RESULTS: Overall, 56.8 percent of dentate adults with diabetes reported having a dental care visit in the preceding year compared with 64.7 percent for those without diabetes. In a multivariable model, diabetes status was significantly associated with having a dental care visit, independent of periodontitis status and covariates. Neither periodontitis status nor sex served as effect modifiers for the association between diabetes status and dental care visits. CONCLUSIONS: These data revealed that dental care visits for dentate adults with diabetes were unrelated to their periodontal health, suggesting that fear of periodontal therapy did not influence visit patterns. These data also showed that dental care visit disparities existed for all adults with diabetes, not just women. Future research should investigate whether factors that are indirectly related to diabetes status, such as competing costs, attitudes, and knowledge, are influencing dental care visit patterns among dentate adults with diabetes. PMID- 19552675 TI - Primary language spoken at home and children's dental service utilization in the United States. AB - OBJECTIVE: Language barriers have been well documented as a contributing factor to disparities in the receipt of medical services, especially for Hispanic children. However, there is a paucity of information on the effect of language barriers on children's dental service utilization. We examined the association of primary language spoken at home with the receipt of preventive and routine dental care for children in the United States. METHODS: We analyzed data from the Medical Expenditure Panel Survey (2002-2004), which contains data on 21,049 children weighted to represent 75.8 million children nationally. RESULTS: Among children aged 1-18 years, 13 percent spoke a language other than English at home. Whites, females, children between the ages of 7 and 12 years, and those whose parents spoke English at home had the highest marginal rates of preventive and routine dental visits. However, the large marginal effect of language, even among Hispanics, was not significant after adjusting for other covariates. Parental education and having a primary provider were the strongest predictors of preventive and routine dental visits. CONCLUSION: Children that did not speak English at home were less likely to receive preventive or routine dental care. However, after adjusting for other socio-economic factors, our study suggests that language barriers may not play as pronounced a role in the receipt of dental care as that documented for medical services. PMID- 19552676 TI - Morphological diagnosis of abetalipoproteinemia and the importance of a freshly prepared peripheral smear. PMID- 19552677 TI - How should the patient with multiple endocrine neoplasia type 1 (MEN 1) be followed? AB - Multiple endocrine neoplasia type 1 (MEN 1) is a complex multi-system disease manifesting a diverse range of primary and secondary metabolic and neoplastic disorders. It is possible to improve patient prognosis by early disease detection and treatment. Regular biochemical and radiological screening for parathyroid, gastro-enteropancreatic, pituitary, intrathorasic and adrenal lesions forms the basis of surveillance. The likelihood of adverse sequelea such as renal and bone disease resulting from hyperparathyroidism, severe peptic ulceration and gastric carcinoidosis secondary to hypergastrinaemia can be ameliorated by early detection and management. PMID- 19552678 TI - Change designation of clotting factors to Arabic numerals. PMID- 19552679 TI - Early activation of coagulation after allergen challenge in patients with allergic asthma. PMID- 19552681 TI - Human alternatively spliced tissue factor is not secreted and does not trigger coagulation. PMID- 19552682 TI - A compound heterozygous mutation in glycoprotein VI in a patient with a bleeding disorder. AB - BACKGROUND: The physiological relevance of the collagen glycoprotein VI (GPVI) receptor was known prior to its recognition as a platelet membrane receptor as several patients lacking GPVI as a consequence of autoantibody inhibition presented with a mild bleeding diathesis. Remarkably, patients with a proven GPVI gene mutation have not yet been identified. RESULTS: In the present study, we describe a patient with a lifelong history of bleeding problems, structurally normal platelets but a functional platelet defect. Platelet aggregations are normal except for an absent response to Horm collagen, convulxin and the collagen related peptide (CRP). ATP dense granule secretion is normal with ADP but absent with Horm collagen. Thrombus formation on a collagen surface in flowing blood is reduced but more single platelets are attached. Remarkably, the platelet function analyzer-100 shows a shortened collagen/ADP closure time. Flow cytometry demonstrates an absent expression of GPVI whereas immunoblot analysis shows strongly reduced levels of GPVI. The patient is compound heterozygous for an out of-frame 16-bp deletion and a missense mutation S175N in a highly conserved residue of the 2nd Ig-like GPVI domain. The parents without clinical bleeding problems are heterozygous carriers. The mother carries the S175N mutation and presents with a mild functional platelet defect. In vitro studies show a reduced membrane expression and convulxin binding with the mutated S175N compared with the wild-type (WT) GPVI receptor. CONCLUSIONS: This study presents the first patient with a proven genetic GPVI defect. PMID- 19552680 TI - Common hemostasis and inflammation gene variants and venous thrombosis in older adults from the Cardiovascular Health Study. AB - BACKGROUND/OBJECTIVES: Age-related changes in blood coagulation and fibrinolysis are associated with increased risk of thrombotic events. Inherited deficiencies of coagulation proteins, such as factor V (FV) Leiden and prothrombin G20210A, explain a small fraction of venous thromboembolic disease (VTE). Additional genetic factors are likely to underlie the etiology of VTE, some of which may become manifest at older ages. METHODS: We tested 290 common SNPs within 51 thrombosis and inflammation genes for association with VTE in the Cardiovascular Health Study, a large, prospective cohort of older adults followed for up to 12 years. RESULTS: There were 184 VTE events that occurred at mean age of 78 years. TagSNPs within four genes encoding FXIII subunit A (F13A), FVII activating protease (HABP2), protease activated receptor-1 (F2R) and the urokinase receptor (PLAUR) showed the strongest evidence for association with VTE, with each gene having a global P-value < 0.05 and at least one tagSNP false discovery rate (FDR) q-value < 0.05. The rs3024409 variant allele of F13A1 was associated with 1.66 fold increased risk of VTE, while the minor alleles of HABP2 rs6585234 and rs3862019, F2R rs253061 and rs153311, and PLAUR rs344782 were each associated with lower risk of VTE (hazard ratios in the range of 0.49-0.66). Consistent with the observed protective association for VTE risk, the HABP2 rs3862019 variant allele was also associated with lower activity levels of coagulation factors FVIII, FIX, FX and plasminogen. We also confirm previously reported associations between common variants of the coagulation FII, FV, FVIII, FXI, alpha-fibrinogen and protein C genes and risk of VTE. CONCLUSIONS: These findings suggest that several novel common coagulation gene variants may be related to risk of VTE in older adults. Further studies in older adults are needed to validate these findings and assess functional molecular mechanisms. PMID- 19552683 TI - Evolution of the fibrinogen gamma' chain: implications for the binding of factor XIII, thrombin and platelets. PMID- 19552684 TI - Safety of ruling out acute pulmonary embolism by normal computed tomography pulmonary angiography in patients with an indication for computed tomography: systematic review and meta-analysis. AB - INTRODUCTION: Several outcome studies have ruled out acute pulmonary embolism (PE) by normal computed tomography pulmonary angiography (CTPA). We performed a meta-analysis in order to determine the safety of this strategy in a specific group of patients with a strict indication for CTPA, that is, 'likely' or 'high' clinical probability for PE, an elevated D-dimer concentration, or both. METHODS: Studies that ruled out PE by normal CTPA, with or without subsequent normal bilateral compression ultrasonography (CUS), in patients with a strict indication for CTPA, were searched for in Medline, EMBASE, Web of Science and the Cochrane dataset. The primary endpoint was the occurrence of (fatal) venous thromboembolism (VTE) in a 3-month follow-up period. RESULTS: Three studies were identified that excluded PE by CTPA alone (2020 patients), and three studies that performed additional CUS of the legs after normal CTPA (1069 patients). The pooled incidence of VTE at 3 months was 1.2% [95% confidence interval (CI) 0.8 1.8] based on a normal CTPA result as a sole test, and 1.1% (95% CI 0.6-2.0) based on normal CTPA and negative CUS findings, resulting in negative predictive values of 98.8% (95% CI 98.2-99.2) and 98.9% (95% CI 98.0-99.4), respectively. This compares favorably with the VTE failure rate after normal pulmonary angiography (1.7%, 95% CI 1.0-2.7). The risk of fatal PE did not differ between the diagnostic strategies (0.6% vs. 0.5%). CONCLUSION: A normal CTPA result alone can safely exclude PE in all patients in whom CTPA is required to rule out this disease. There is no need for additional ultrasonography to rule out VTE in these patients. PMID- 19552686 TI - Variability of anti-PF4/heparin antibody results obtained by the rapid testing system ID-H/PF4-PaGIA: a rebuttal. PMID- 19552688 TI - Looking forwards or looking backwards in avian phylogeography? A comment on Zink and Barrowclough 2008. PMID- 19552689 TI - Actions of veratridine on tetrodotoxin-sensitive voltage-gated Na currents, Na1.6, in murine vas deferens myocytes. AB - BACKGROUND AND PURPOSE: The effects of veratridine, an alkaloid found in Liliaceae plants, on tetrodotoxin (TTX)-sensitive voltage-gated Na(+) channels were investigated in mouse vas deferens. EXPERIMENTAL APPROACH: Effects of veratridine on TTX-sensitive Na(+) currents (I(Na)) in vas deferens myocytes dispersed from BALB/c mice, homozygous mice with a null allele of Na(V)1.6 (Na(V)1.6(-/-)) and wild-type mice (Na(V)1.6(+/+)) were studied using patch-clamp techniques. Tension measurements were also performed to compare the effects of veratridine on phasic contractions in intact tissues. KEY RESULTS: In whole-cell configuration, veratridine had a concentration-dependent dual action on the peak amplitude of I(Na): I(Na) was enhanced by veratridine (1-10 microM), while higher concentrations (> or =30 microM) inhibited I(Na). Additionally, two membrane current components were evoked by veratridine, namely a sustained inward current during the duration of the depolarizing rectangular pulse and a tail current at the repolarization. Although veratridine caused little shift of the voltage dependence of the steady-state inactivation curve and the activation curve for I(Na), veratridine enhanced a non-inactivating component of I(Na). Veratridine caused no detectable contractions in vas deferens from Na(V)1.6(-/-) mice, although in tissues from Na(V)1.6(+/+) mice, veratridine (> or =3 microM) induced TTX-sensitive contractions. Similarly, no detectable inward currents were evoked by veratridine in Na(V)1.6(-/-) vas deferens myocytes, while veratridine elicited both the sustained and tail currents in cells taken from Na(V)1.6(+/+) mice. CONCLUSIONS AND IMPLICATIONS: These results suggest that veratridine possesses a dual action on I(Na) and that the veratridine-induced activation of contraction is induced by the activation of Na(V)1.6 channels. PMID- 19552690 TI - Silibinin prevents amyloid beta peptide-induced memory impairment and oxidative stress in mice. AB - BACKGROUND AND PURPOSE: Accumulated evidence suggests that oxidative stress is involved in amyloid beta (Abeta)-induced cognitive dysfunction. Silibinin (silybin), a flavonoid derived from the herb milk thistle (Silybum marianum), has been shown to have antioxidative properties; however, it remains unclear whether silibinin improves Abeta-induced neurotoxicity. In the present study, we examined the effect of silibinin on the memory impairment and accumulation of oxidative stress induced by Abeta(25-35) in mice. EXPERIMENTAL APPROACH: Aggregated Abeta(25-35) (3 nmol) was intracerebroventricularly administered to mice. Treatment with silibinin (2, 20 and 200 mg.kg(-1), once a day, p.o.) was started immediately after the injection of Abeta(25-35). Locomotor activity was evaluated 6 days after the Abeta(25-35) treatment, and cognitive function was evaluated in a Y-maze and novel object recognition tests 6-11 days after the Abeta(25-35) treatment. The levels of lipid peroxidation (malondialdehyde) and antioxidant (glutathione) in the hippocampus were measured 7 days after the Abeta(25-35) injection. KEY RESULTS: Silibinin prevented the memory impairment induced by Abeta(25-35) in the Y-maze and novel object recognition tests. Repeated treatment with silibinin attenuated the Abeta(25-35)-induced accumulation of malondialdehyde and depletion of glutathione in the hippocampus. CONCLUSIONS AND IMPLICATIONS: Silibinin prevents memory impairment and oxidative damage induced by Abeta(25-35) and may be a potential therapeutic agent for Alzheimer's disease. PMID- 19552691 TI - Functional evidence for the expression of P2X1, P2X4 and P2X7 receptors in human lung mast cells. AB - BACKGROUND AND PURPOSE: P2X receptors are widely expressed in cells of the immune system with varying functions. This study sought to characterize P2X receptor expression in the LAD2 human mast cell line and human lung mast cells (HLMCs). EXPERIMENTAL APPROACH: Reverse transcriptase polymerase chain reaction (RT-PCR) and patch clamp studies were used to characterize P2X expression in mast cells using a range of pharmacological tools. KEY RESULTS: RT-PCR revealed P2X1, P2X4 and P2X7 transcripts in both cell types; mRNA for P2X6 was also detected in LAD2 cells. Under whole-cell patch clamp conditions, rapid application of ATP (1-1000 microM) to cells clamped at -60 mV consistently evoked inward currents in both types of cells. Brief application of ATP (1 s) evoked a rapidly desensitizing P2X1-like current in both cell types. This current was also elicited by alphabetamethylene ATP (10 microM, 94% cells, n= 31) and was antagonized in LAD2 cells by NF 449 (1 microM) and pyridoxal phosphate-6-azo(benzene-2,4-disulphonic acid) (1-10 microM). A P2X7-like non-desensitizing current in response to high concentrations of ATP (1-5 mM) was also seen in both cell types (96% LAD2, n= 24; 54% HLMCs, n= 24) which was antagonized by AZ11645373 (1 microM). P2X7-like responses were also evoked in LAD2 cells by 2'(3')-0-(4-benzoylbenzoyl)ATP (300 microM). A P2X4-like current was evoked by 100 microM ATP (80% LAD2, n= 10; 21% HLMCs, n= 29), the amplitude and duration of which was potentiated by ivermectin (3 microM). CONCLUSION AND IMPLICATIONS: Our data confirmed the presence of functional P2X1, P2X4 and P2X7 receptors in LAD2 cells and HLMCs. PMID- 19552693 TI - Phenotypic selection on leaf ecophysiological traits in Helianthus. AB - Habitats that differ in soil resource availability are expected to differ for selection on resource-related plant traits. Here, we examined spatial and temporal variation in phenotypic selection on leaf ecophysiological traits for 10 Helianthus populations, including two species of hybrid origin, Helianthus anomalus and Helianthus deserticola, and artificial hybrids of their ancestral parents. Leaf traits assessed were leaf size, succulence, nitrogen (N) concentration and water-use efficiency (WUE). Biomass and leaf traits of artificial hybrids indicate that the actively moving dune habitat of H. anomalus was more growth limiting, with lower N availability but higher relative water availability than the stabilized dune habitat of H. deserticola. Habitats differed for direct selection on leaf N and WUE, but not size or succulence, for the artificial hybrids. However, within the H. anomalus habitat, direct selection on WUE also differed among populations. Across years, direct selection on leaf traits did not differ. Leaf N was the only trait for which direct selection differed between habitats but not within the H. anomalus habitat, suggesting that nutrient limitation is an important selective force driving adaptation of H. anomalus to the active dune habitat. PMID- 19552692 TI - Constitutive activity of cannabinoid-2 (CB2) receptors plays an essential role in the protean agonism of (+)AM1241 and L768242. AB - BACKGROUND AND PURPOSE: Cannabinoid-2 (CB(2)) receptor-selective agonists have shown anti-nociceptive activity in models of neuropathic and inflammatory pain, and the two agonists most widely used, (+/-)AM1241 [(2-iodo-5-nitrophenyl)-[1-(1 methylpiperidin-2-ylmethyl)-1H-indol-3-yl-methanone] and L768242 [(2,3-dichloro phenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-yl-ethyl)-indol-1-yl]-methanone] (GW405833), have been suggested to be protean agonists. Here we investigated the role of the constitutive activity of CB(2) receptors in (+)AM1241 and L768242 protean agonism. EXPERIMENTAL APPROACH: Pharmacological profiles of CB(2) receptor ligands were evaluated in Chinese hamster ovary cells expressing recombinant human (hCB(2)) or rat (rCB(2)) receptors, by measuring modulation of cAMP. To assess the influence of constitutive activity on pharmacological profile, constitutive activity was abolished by pretreatment with AM630 [(6-iodo 2-methyl-1-[2-(4-morpholinyl)ethyl]-1H-indol-3-yl](4-methoxyphenyl) methanone)], followed by extensive washing. KEY RESULTS: In cell lines expressing either hCB(2) or rCB(2) receptors, (+)AM1241 did not reverse forskolin stimulation of cAMP levels. Conversely, L768242 was an inverse agonist at both hCB(2) and rCB(2) receptors. Abolition of constitutive activity disclosed (+)AM1241 and L768242 agonist activity, while activity of CP55940 [5-(1,1-dimethylheptyl)-2-[(1R,2R,5R) 5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-phenol] was unaffected and AM630 became a neutral antagonist. In presence of constitutively active CB(2) receptors, (+)AM1241 antagonized CP55940, but when constitutive activity was abolished, it acted as a partial agonist with additive or antagonistic behaviour, depending on concentration. CONCLUSIONS AND IMPLICATIONS: These results show that (+)AM1241 and L768242 are protean agonists at both hCB(2) and rCB(2) receptors. Abolition of constitutive activity reveals the agonist activity of these compounds. Thus, differences between in vivo and in vitro profiles of CB(2) receptor agonists could be due to different levels of constitutive activity in recombinant versus native CB(2) receptors. PMID- 19552694 TI - Darwin's beautiful contrivances: evolutionary and functional evidence for floral adaptation. AB - Although not 'a professed botanist', Charles Darwin made seminal contributions to understanding of floral and inflorescence function while seeking evidence of adaptation by natural selection. This review considers the legacy of Darwin's ideas from three perspectives. First, we examine the process of floral and inflorescence adaptation by surveying studies of phenotypic selection, heritability and selection responses. Despite widespread phenotypic and genetic capacity for natural selection, only one-third of estimates indicate phenotypic selection. Second, we evaluate experimental studies of floral and inflorescence function and find that they usually demonstrate that reproductive traits represent adaptations. Finally, we consider the role of adaptation in floral diversification. Despite different diversification modes (coevolution, divergent use of the same pollen vector, pollinator shifts), evidence of pollination ecotypes and phylogenetic patterns suggests that adaptation commonly contributes to floral diversity. Thus, this review reveals a contrast between the inconsistent occurrence of phenotypic selection and convincing experimental and comparative evidence that floral traits are adaptations. Rather than rejecting Darwin's hypotheses about floral evolution, this contrast suggests that the tempo of creative selection varies, with strong, consistent selection during episodes of diversification, but relatively weak and inconsistent selection during longer, 'normal' periods of relative phenotypic stasis. PMID- 19552695 TI - Exploding a myth: the capsule dehiscence mechanism and the function of pseudostomata in Sphagnum. AB - The nineteenth century air-gun explanation for explosive spore discharge in Sphagnum has never been tested experimentally. Similarly, the function of the numerous stomata ubiquitous in the capsule walls has never been investigated. Both intact and pricked Sphagnum capsules, that were allowed to dry out, all dehisced over an 8-12 h period during which time the stomatal guard cells gradually collapsed and their potassium content, measured by X-ray microanalysis in a cryoscanning electron microscope, gradually increased. By contrast, guard cell potassium fell in water-stressed Arabidopsis. The pricking experiments demonstrate that the air-gun notion for explosive spore discharge in Sphagnum is inaccurate; differential shrinkage of the capsule walls causes popping off the rigid operculum. The absence of evidence for a potassium-regulating mechanism in the stomatal guard cells and their gradual collapse before spore discharge indicates that their sole role is facilitation of sporophyte desiccation that ultimately leads to capsule dehiscence. Our novel functional data on Sphagnum, when considered in relation to bryophyte phylogeny, suggest the possibility that stomata first appeared in land plants as structures that facilitated sporophyte drying out before spore discharge and only subsequently acquired their role in the regulation of gaseous exchange. PMID- 19552696 TI - Duffy antigen expression on reticulocytes does not alter following blood loss in an autologous donation model. AB - BACKGROUND: The Duffy blood group (Fy) antigen functions as the receptor whereby the malarial parasite Plasmodium vivax invades reticulocytes. In this study, we evaluated an autologous blood donation model to measure Fy expression during the anticipated response to blood loss. AIMS: This study aims to examine Fy expression following anticipated reticulocytosis in response to blood loss from autologous whole blood donation. METHOD: Subjects were healthy blood donors presenting for planned collection of two or three autologous units. Whole blood (450 ml +/- 10%) was collected and processed. Blood samples for Fy testing were obtained from the donations. These were assayed by flow cytometry by measuring binding of a phycoerythrin-labelled anti-Fy6 antibody and compared against reticulocyte numbers. Reticulocyte numbers were measured using thiazole orange. Results were compared from baseline (first donation) with samples at second and, if available, third, donations. Phenotyping for Fy a and b antigens was performed. RESULTS: Reticulocytes increased by a mean of 37% over baseline [0.93% (range 0.31-1.93) to 1.23% (0.32-3.51%)] following donation of two (n = 32) or three (n = 9) autologous whole blood units. Absolute reticulocyte count remained low. Mean and median Fy expression on mature red blood cells and reticulocytes did not change from baseline levels despite individual variation. No apparent relationship to serologically determined Fy a and/or b antigen status was present. CONCLUSION: Baseline expression of Fy antigen on mature red blood cells and reticulocytes is quite variable between individuals, but appears not to be greatly affected by mild to moderate reticulocytosis following blood loss in an autologous blood donation model. PMID- 19552697 TI - Sialylated ligands on pathogenic Trypanosoma cruzi interact with Siglec-E (sialic acid-binding Ig-like lectin-E). AB - Trypanosoma cruzi causes a suppression of the immune system leading to persistence in host cells. The trans-sialidase expressed by T. cruzi is a major virulence factor and transfers sialic acid from host glycoconjugates to mucin like molecules on the parasite. Here we demonstrate that these sialylated structures play a role in the immunosuppression. We used two T. cruzi strains, whose TS activity correlated with their pathogenicity. The Tulahuen strain, characterized by a high TS activity efficiently infected mice, whereas the Tehuantepec strain showing a reduced TS activity could not establish a patent parasitemia. In vitro analysis revealed that these two strains invaded phagocytic and non-phagocytic host cells at a comparable rate, but they exhibited different potentials to modulate dendritic cell function. In contrast to Tehuantepec, the Tulahuen strain suppressed the production of the proinflammatory cytokine IL-12 and subsequent T-cell activation. This inhibitory effect was absent upon desialylation of the parasite. Therefore, we analysed whether sialylated structures of T. cruzi interact with the inhibitory sialic acid-binding protein Siglec-E on DC. Indeed, Siglec-E interacted with the pathogenic Tulahuen strain, but showed a diminished binding to the Tehuantepec strain. Ligation of Siglec-E on DC using antibodies confirmed this inhibitory effect on DC function. PMID- 19552699 TI - CNS and lung metastasis of sebaceous epithelioma in a dog. AB - Sebaceous epithelioma is a common canine cutaneous neoplasm characterized by a preponderance of basaloid cells with few well-differentiated sebocytes. It is considered a low-grade malignancy, as it may occasionally recur at the excision site; the possibility of lymph node metastasis is anecdotally reported, and distant metastases have never been observed. This case report presents the clinical and pathological features of a sebaceous epithelioma of the upper lip with a highly aggressive behaviour. The patient was a 9-year-old female dachshund that developed local recurrence 11 months after the excision of the primary sebaceous epithelioma and multiple lung and central nervous system metastases 5 months later. The designation epitheliomatous sebaceous carcinoma has been suggested for aggressive sebaceous epitheliomas, although differential criteria are still to be determined. PMID- 19552700 TI - A systematic review of the evidence of reduced allergenicity and clinical benefit of food hydrolysates in dogs with cutaneous adverse food reactions. AB - Several hydrolysate-based diets have been commercialized for helping diagnose or treat dogs with cutaneous adverse food reactions (CAFR). This systematic review was performed to examine the evidence in favour of reduced immunological and clinical allergenicity of hydrolysates in dogs with CAFR. Citation databases, meeting abstracts and article bibliographies were scanned for relevant citations, and companies were contacted to provide unpublished reports. Eleven studies relevant to this study were identified. Some evidence of reduced serum IgE binding to a soy hydrolysate (1 study) and decreased intradermal test reactivity to hydrolysed proteins (three studies) was found. In four reports, the feeding of dogs suspected of having CAFR with hydrolysate-based diets reduced or eliminated clinical signs in a variable proportion of subjects. The percentage of dogs with CAFR that still reacted to these hydrolysate-based diets could not be assessed, however. Importantly, up to 50% of dogs with CAFR enrolled in three controlled studies exhibited increases in clinical signs after ingesting partial hydrolysates derived from foods to which they were hypersensitive. In conclusion, the limited number of studies undertaken point to reduced - but not eliminated - immunological and clinical allergenicity of hydrolysate-based commercial diets. A variable proportion of dogs with CAFR will exhibit a worsening of clinical signs when fed partial hydrolysates. Clinicians must weigh the clinical benefit of these diets versus their high cost and low risk of reduced appetence or gastrointestinal sign development. At this time, hydrolysate-containing diets are probably best used in dogs suspected not to be hypersensitive to their individual components. PMID- 19552698 TI - Human Toll-like receptor 4 responses to P. gingivalis are regulated by lipid A 1- and 4'-phosphatase activities. AB - Signal transduction following binding of lipopolysaccharide (LPS) to Toll-like receptor 4 (TLR4) is an essential aspect of host innate immune responses to infection by Gram-negative pathogens. Here, we describe a novel molecular mechanism used by a prevalent human bacterial pathogen to evade and subvert the human innate immune system. We show that the oral pathogen, Porphyromonas gingivalis, uses endogenous lipid A 1- and 4'-phosphatase activities to modify its LPS, creating immunologically silent, non-phosphorylated lipid A. This unique lipid A provides a highly effective mechanism employed by this bacterium to evade TLR4 sensing and to resist killing by cationic antimicrobial peptides. In addition, lipid A 1-phosphatase activity is suppressed by haemin, an important nutrient in the oral cavity. Specifically, P. gingivalis grown in the presence of high haemin produces lipid A that acts as a potent TLR4 antagonist. These results suggest that haemin-dependent regulation of lipid A 1-dephosphorylation can shift P. gingivalis lipid A activity from TLR4 evasive to TLR4 suppressive, potentially altering critical interactions between this bacterium, the local microbial community and the host innate immune system. PMID- 19552701 TI - Tepoxalin reduces pruritus and modified CADESI-01 scores in dogs with atopic dermatitis: a prospective, randomized, double-blinded, placebo-controlled, cross over study. AB - Thirty dogs with atopic dermatitis were given tepoxalin (Zubrin, Intervet/Schering-Plough Animal Health, Boxmeer, the Netherlands) or placebo once daily for 4 weeks, followed by a wash-out period of 1 week before reversing the treatments. Pruritus was scored by the owners using the Edinburgh Pruritus Scale and one investigator employed a modification of the Canine Atopic Dermatitis Extent and Severity Index-01 (mCADESI-01) to score the physical lesions. After administration of tepoxalin there was a > or = 50% reduction in pruritus and mCADESI-01 scores in 36% and 25% of the dogs, respectively, whereas following administration of the placebo there was a > or = 50% reduction in pruritus and mCADESI-01 scores in only 25% and 16% of the dogs, respectively. Analysis of pooled data indicated that tepoxalin resulted in a significant reduction in pruritus (P = 0.012) and mCADESI-01 (P = 0.002) scores but there was no significant change after placebo. The median pruritus scores before and after tepoxalin were 2 (range 1-5) and 1 (range 0-5), respectively, and before and after placebo were 2 (range 0-4) and 2 (range 0-4), respectively. The median mCADESI scores before and after tepoxalin were 23 (range 0-68) and 16 (range 0 72), respectively, and before and after placebo were 18 (range 3-79) and 24 (range 0-65), respectively. At the dose used in this study (10.0-19.1 mg kg(-1)), tepoxalin was well-tolerated and no adverse effects were noted. PMID- 19552702 TI - CNS arousal and neurobehavioral performance in a short-term sleep restriction paradigm. AB - Few studies have investigated waking electrophysiological measures of arousal during sleep restriction. This study examined electroencephalogram (EEG) activity and performance during a 96-hour laboratory protocol where participants slept a baseline night (8 h), were randomly assigned to 3-, 5-, or 8-hour sleep groups for the next two nights sleep restriction (SR1, SR2), and then slept a recovery night (8 h). There were dose-dependent deficits on measures of mood, sleepiness, and reaction time that were apparent during this short-term bout of sleep restriction. The ratio of alpha to theta EEG recorded at rest indicated dose dependent changes in CNS arousal. At 9:00 hours, both the 3- and 5-hour groups showed EEG slowing (sleepiness) during restriction, with the 3-hour group exhibiting greater deficits. Later in the day at 13:00 hours, the 5-hour group no longer exhibited EEG slowing, but the extent of slowing was more widespread across the scalp for the 3-hour group. High-frequency EEG, a measure of effort, was greater on the mornings following sleep restriction. The 5-hour group had increased beta EEG at central-parietal sites following both nights of restriction, whereas the 3-hour group had increased beta and gamma EEG at occipital regions following the first night only. Short-term sleep restriction leads to deficits in performance as well as EEG slowing that correspond to the amount and duration of sleep loss. High-frequency EEG may be a marker of effort or compensation. PMID- 19552703 TI - Information processing during sleep: the effect of olfactory stimuli on dream content and dream emotions. AB - Research has shown that external stimuli presented during sleep can affect dream content, thus reflecting information processing of the sleeping brain. Olfactory stimuli should have a stronger effect on dream emotions because their processing is linked directly to the limbic system. Because selective olfactory stimulation does not increase arousal activity, intense olfactory stimulation is therefore a prime paradigm for studying information processing during sleep. Fifteen healthy, normosmic volunteers were studied by intranasal chemosensory stimulation during rapid eye movement sleep based on air-dilution olfactometry. For olfactory stimulation, hydrogen sulphide (smell of rotten eggs) and phenyl ethyl alcohol (smell of roses) was used and compared with a control condition without stimulation. The olfactory stimuli affected significantly the emotional content of dreams: the positively toned stimulus yielded more positively toned dreams, whereas the negative stimulus was followed by more negatively toned dreams. Direct incorporations, i.e. the dreamer is smelling something, were not found. The findings indicate that information processing of olfactory stimuli is present in sleep and that the emotional tone of dreams can be influenced significantly depending upon the hedonic characteristic of the stimulus used. It would be interesting to conduct learning experiments (associating specific odours with declarative material) to study whether this declarative material is incorporated into subsequent dreams if the corresponding odour cue is presented during sleep. It would also be interesting to study the effect of positively toned olfactory stimuli on nightmares. PMID- 19552704 TI - Sleep and sleep homeostasis in constant darkness in the rat. AB - According to the two-process model of sleep regulation, a homeostatic Process S increases during waking and decreases during sleep. The time course of Process S can be derived on the basis of changes in vigilance states and changes in electroencephalogram slow-wave activity (SWA, activity below 4 Hz) during non rapid eye movement (NREM) sleep. In most mouse strains, an optimal fit between S and SWA was achieved with one increasing (active during waking and REM sleep) and one decreasing time constant (active during NREM sleep) for Process S. However, in the rat, systematic deviations in the light and dark periods were observed, which were resolved by introducing different decreasing time constants between the light and dark periods. The present study shows that this difference between the rest (light) and active (dark) phases remains, and may even be larger, after animals are adapted to constant dark conditions for at least a week. In addition, the data show that the build-up rate of SWA at the onset of a NREM sleep episode is slow compared with the increase rate under light-dark conditions, and that this build-up rate changes with the circadian phase. The slow build-up rate introduces a systematic error between the simulation of Process S and SWA in NREM sleep. The circadian modulation of the build-up rate may, together with circadian changes in NREM sleep episode duration, be the source of the necessity of introducing a difference in the decreasing time constant between the rest and active phases. PMID- 19552705 TI - A phenotypic study of TFS1 mutants differentially altered in the inhibition of Ira2p or CPY. AB - The Saccharomyces cerevisiae protein Tfs1p is known as a dual protein. On the one hand, it inhibits the carboxypeptidase Y protease, and on the other, it inhibits Ira2p, a GTPase-activating protein of Ras. We managed to dissect precise areas of Tfs1p specifically involved in only one of those functions. Based on these data, specific Tfs1p point mutants affected in only one of these two functions were constructed. In order to obtain insights on the physiological role of these functions, systematic phenotypic tests were performed on strains expressing these specific Tfs1p mutants. The results obtained demonstrate that the inhibition of Ira2p by Tfs1p is the predominant function under the conditions tested. PMID- 19552707 TI - The PagN protein mediates invasion via interaction with proteoglycan. AB - Heparan sulphate proteoglycans are major components of the mammalian cell membrane. Here we show that PagN of Salmonella enterica serovar Typhimurium utilizes heparinated proteoglycan to successfully invade mammalian cells. Mutants defective in the production of the outer membrane protein PagN displayed similar levels of invasiveness of glycosylation-deficient pgsA-745 cells in comparison with wild-type Salmonella. Furthermore, pgsA-745 cells were invaded c. 400-fold less efficiently than CHO-K1 cells by Escherichia coli expressing PagN. PagN is likely to interact with heparinated proteoglycan as heparin could inhibit PagN mediated invasion in a dose-dependent manner. Finally, we show, by deletion analysis, that all four extracellular loops of PagN are crucial for invasion of mammalian cells. PMID- 19552706 TI - Cross-kingdom interactions: Candida albicans and bacteria. AB - Bacteria and fungi are found together in a myriad of environments and particularly in a biofilm, where adherent species interact through diverse signaling mechanisms. Yet, despite billions of years of coexistence, the area of research exploring fungal-bacterial interactions, particularly within the context of polymicrobial infections, is still in its infancy. However, reports describing a multitude of wide-ranging interactions between the fungal pathogen Candida albicans and various bacterial pathogens are on the rise. An example of a mutually beneficial interaction is coaggregation, a phenomenon that takes place in oral biofilms where the adhesion of C. albicans to oral bacteria is considered crucial for its colonization of the oral cavity. In contrast, the interaction between C. albicans and Pseudomonas aeruginosa is described as being competitive and antagonistic in nature. Another intriguing interaction is that occurring between Staphylococcus aureus and C. albicans, which although not yet fully characterized, appears to be initially synergistic. These complex interactions between such diverse and important pathogens would have significant clinical implications if they occurred in an immunocompromised host. Therefore, understanding the mechanisms of adhesion and signaling involved in fungal bacterial interactions may lead to the development of novel therapeutic strategies for impeding microbial colonization and development of polymicrobial disease. PMID- 19552708 TI - Glutathione production by efficient ATP-regenerating Escherichia coli mutants. AB - There is an ongoing demand to improve the ATP-regenerating system for industrial ATP-driven bioprocesses because of the low efficiency of ATP regeneration. To address this issue, we investigated the efficiency of ATP regeneration in Escherichia coli using the Permeable Cell Assay. This assay identified 40 single gene deletion strains that had over 150% higher total cellular ATP synthetic activity relative to the parental strain. Most of them also showed higher ATP driven glutathione synthesis. The deleted genes of the identified strains that showed increased efficiency of ATP regeneration for glutathione production could be divided into the following four groups: (1) glycolytic pathway-related genes, (2) genes related to degradation of ATP or adenosine, (3) global regulatory genes, and (4) genes whose contribution to the ATP regeneration is unknown. Furthermore, the high glutathione productivity of DeltanlpD, the highest glutathione-producing mutant strain, was due to its reduced sensitivity to the externally added ATP for ATP regeneration. This study showed that the Permeable Cell Assay was useful for improving the ATP-regenerating activity of E. coli for practical applications in various ATP-driven bioprocesses, much as that of glutathione production. PMID- 19552709 TI - DivIVA uses an N-terminal conserved region and two coiled-coil domains to localize and sustain the polar growth in Corynebacterium glutamicum. AB - Corynebacterium glutamicum is a rod-shaped actinomycete with a distinct model of peptidoglycan synthesis during cell elongation, which takes place at the cell poles and is sustained by the essential protein DivIVA(CG) (C. glutamicum DivIVA). This protein contains a short conserved N-terminal domain and two coiled coil regions: CC1 and CC2. Domain deletions and chimeric versions of DivIVA were used to functionally characterize the three domains, and all three were found to be essential for proper DivIVA(CG) function. However, in the presence of the N terminal domain from DivIVA(CG), either of the two coiled-coil domains of DivIVA(CG) could be replaced by the equivalent coiled-coil domain of Bacillus subtilis DivIVA (DivIVA(BS)) without affecting the function of the original DivIVA(CG), and more than one domain had to be exchanged to lose function. Although no single domain was sufficient for subcellular localization or function, CC1 was mainly implicated in stimulating polar growth and CC2 in targeting to DivIVA(CG) assemblies at the cell poles in C. glutamicum. PMID- 19552710 TI - Domains involved in the in vivo function and oligomerization of apical growth determinant DivIVA in Streptomyces coelicolor. AB - The coiled-coil protein DivIVA is a determinant of apical growth and hyphal branching in Streptomyces coelicolor. We have investigated the properties of this protein and the involvement of different domains in its essential function and subcellular targeting. In S. coelicolor cell extracts, DivIVA was present as large oligomeric complexes that were not strongly membrane associated. The purified protein could self-assemble into extensive protein filaments in vitro. Two large and conspicuous segments in the amino acid sequence of streptomycete DivIVAs not present in other homologs, an internal PQG-rich segment and a carboxy terminal extension, are shown to be dispensable for the essential function in S. coelicolor. Instead, the highly conserved amino-terminal of 22 amino acids was required and affected establishment of new DivIVA foci and hyphal branches, and an essential coiled-coil domain affected oligomerization of the protein. PMID- 19552711 TI - Contribution of phosphoglucosamine mutase to the resistance of Streptococcus gordonii DL1 to polymorphonuclear leukocyte killing. AB - Phosphoglucosamine mutase (GlmM; EC 5.4.2.10) catalyzes the interconversion of glucosamine-6-phosphate to glucosamine-1-phosphate, an essential step in the biosynthetic pathway leading to the formation of the peptidoglycan precursor uridine 5'-diphospho-N-acetylglucosamine. We have recently identified the gene (glmM) encoding the enzyme of Streptococcus gordonii, an early colonizer on the human tooth and an important cause of infective endocarditis, and indicated that the glmM mutation in S. gordonii appears to influence bacterial cell growth, morphology, and sensitivity to penicillins. In the present study, we assessed whether the glmM mutation also affects escape from polymorphonuclear leukocyte (PMN)-dependent killing. Although no differences in attachment to human PMNs were observed between the glmM mutant and the wild-type S. gordonii, the glmM mutation resulted in increased sensitivity to PMN-dependent killing. Compared with the wild type, the glmM mutant induced increased superoxide anion production and lysozyme release by PMNs. Moreover, the glmM mutant is more sensitive to lysozyme, indicating that the GlmM may be required for synthesis of firm peptidoglycans for resistance to bacterial cell lysis. These findings suggest that the GlmM contributes to the resistance of S. gordonii to PMN-dependent killing. Enzymes such as GlmM could be novel drug targets for this organism. PMID- 19552712 TI - Possible promoter regions within the proteolytic system in Streptococcus thermophilus and their interaction with the CodY homolog. AB - Possible promoter regions preceding 14 genes belonging to the proteolytic system of Streptococcus thermophilus KLDS 3.0503 were predicted by a promoter analysis software nnpp. The 14 genes included an extracellular protease gene prtS, an oligopeptide ABC transport system gene amiA1, and 12 genes, respectively, encoding peptidases pepA, pepS, pepN, pepC, pepB, pepQ, pepV, pepT, pepM, pepXP, pepP, and pepO. These predicted promoter sequences were cloned and inserted into the upstream of a promoterless Escherichia coli gusA (beta-glucuronidase) gene in a promoter probe vector pNZ273. The resulting vectors were, respectively, introduced into S. thermophilus KLDS 3.0503 and all 14 predicted promoter sequences were able to drive gusA expression, which indicated that these sequences were functional promoters. These promoters were able to interact with the S. thermophilus CodY homolog in an in vitro DNA binding assay but they did not contain a conserved CodY-box sequence identified in Lactococcus lactis. These results were useful for further studies on the regulation of protein metabolism in S. thermophilus. PMID- 19552713 TI - Ferredoxin-NADP reductase from the thermophilic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6. AB - The thermophilic, obligately chemolithoautotrophic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6, assimilates carbon dioxide via the reductive tricarboxylic acid cycle. Small iron-sulfur proteins, ferredoxins, play a central role as low-potential electron donors for this cycle. The fpr gene of this bacterium, encoding a putative ferredoxin-NADP(+) reductase (FNR, EC 1.18.1.2), was expressed in Escherichia coli, and the recombinant protein was purified to homogeneity. Unexpectedly, the monomeric Fpr protein contained one molecule of FMN as a prosthetic group, although FNRs from other organisms are known to contain FAD. The FMN-containing Fpr was shown to be a bona fide FNR that catalyzes a reversible redox reaction between NADP(+)/NADPH and ferredoxins. PMID- 19552715 TI - Anti-inflammatory activity of a globular adiponectin function on RAW 264 cells stimulated by lipopolysaccharide from Aggregatibacter actinomycetemcomitans. AB - Adiponectin is an adipokine with potent anti-inflammatory properties. We previously reported that a globular adiponectin (gAd) suppresses Aggregatibacter actinomycetemcomitans lipopolysaccharide-induced nuclear factor-kappaB activity, suggesting an anti-inflammatory effect of gAd. In this study, we investigated whether gAd is able to modulate the effect of A. actinomycetemcomitans lipopolysaccharide on cytokine induction in a murine macrophage cell line (RAW 264). The phosphorylation of p38 mitogen-activated protein kinase, c-Jun N terminal kinase, extracellular signal-regulated kinase, and IkappaB kinase alpha/beta and the degradation of IkappaB, which were induced by A. actinomycetemcomitans lipopolysaccharide intoxication, were clearly reduced in gAd-pretreated RAW 264 cells compared with the untreated cells. Expression levels of tumor necrosis factor (TNF)-alpha and interleukin-10 (IL-10) mRNA were assessed by real-time PCR. Cell-free supernatants were collected after 12 h of stimulation and analyzed by enzyme-linked immunosorbent assay for TNF-alpha and IL-10. Pretreatment with gAd significantly inhibited the A. actinomycetemcomitans lipopolysaccharide-induced TNF-alpha mRNA expression and protein secretion. In contrast, pretreatment with gAd significantly enhanced the A. actinomycetemcomitans lipopolysaccharide-induced IL-10 mRNA expression and protein secretion. These data suggest a mechanism for the anti-inflammatory activity of gAd in local inflammatory lesions, such as periodontitis. PMID- 19552716 TI - Low-dose methotrexate treatment for moderate-to-severe atopic dermatitis in adults. AB - BACKGROUND: Atopic dermatitis (AD) is a common inflammatory skin disease. Methotrexate (MTX) was suggested as an effective treatment option in cases of moderate-to-severe atopic dermatitis. This study assessed the efficacy and safety of treatment with low weekly doses of methotrexate for moderate-to-severe AD in adults. METHODS: Twenty adult patients with moderate-to-severe AD were included in this retrospective study. Those patients were unresponsive to topical treatments, antihistamines and at least one of the second-line treatments. MTX in low weekly doses of 10-25 mg was administered orally or intramuscularly with folic acid supplementation 5 mg per week for at least 8-12 weeks. The response to treatment was evaluated by change in SCORAD (SCORing Atopic Dermatitis), DLQI (Dermatology Quality of Life Index) and the global assessment of the clinical response score. RESULTS: After 8-12 weeks of treatment, we observed an objective response in most patients. There were 16 responders and 4 non-responders. The mean SCORAD and DLQI decreased by 28.65 units (44.3%) and 10.15 units (43.5%), respectively. The first improvement was observed after a period ranging from 2 weeks to 3 months (mean 9.95 w +/- 3.17). Treatment was more effective in adult onset AD than in childhood onset. Tolerance of treatment was good. However, nausea and an increase of liver enzymes were observed in 5 patients and 3 of them required a transient discontinuation of MTX. One patient developed peripheral neuropathy, which was resolved several weeks after the discontinuation of MTX. CONCLUSION: MTX seems to be an effective and safe second-line treatment for patients with moderate-to-severe atopic dermatitis. A randomized, controlled study is warranted. PMID- 19552717 TI - The influence of treatment on quality of life in systemic lupus erythematosus patients. AB - OBJECTIVE: Systemic lupus erythematosus is an autoimmune disease with uncertain prognosis as to the course. The patients need pharmacotherapy all their life. The main aim of this study was to determinate the impact of therapeutic schedules on patients' quality of life. METHODS: The study was performed on 83 patients who were divided into five groups according to methods of treatment used. Quality of life was measured using MOS SF-36. RESULTS: Obtained results revealed that therapeutic schedules have the highest influence on patients' social functioning. The more medicaments the patients use, the lower their quality of life. CONCLUSIONS: The type of medicaments and therapeutic schemes adopted affect the patients' quality of life. Nevertheless, quality of life and the patients' attitude to it are very subjective and are also influenced by the clinical state of the patients as well as many other factors such as socioeconomic and demographic. PMID- 19552718 TI - Expression of human neutrophil proteins in acne vulgaris. AB - BACKGROUND: In acne vulgaris patients, the presence of a dysregulation of the production of innate and specific antimicrobial peptides has been postulated. OBJECTIVE: This study aims to determine whether human neutrophil proteins (HNP) 1 3 are expressed in acne patients. MATERIALS AND METHODS: HNP 1-3 expression was investigated in 35 acne patients treated with isotretinoin and in 25 healthy subjects. At the beginning of the study, two skin biopsies were taken from acne patients; one biopsy was taken from an established pustule and one from uninvolved skin, and the biopsies were repeated after treatment. Only one biopsy was obtained from controls. RESULTS: The statistical analysis showed that pustular lesions of acne patients had significantly higher levels of perivascular and interstitial HNP 1-3 expression when compared with the biopsy of uninvolved skin of these patients (P = 0.003, P = 0.001, respectively) and with that of healthy controls (P = 0.007, P = 0.014, respectively). Isotretinoin treatment achieved a decrease in the perivascular and interstitial HNP 1-3 expression of pustular lesions (P = 0.01, P = 0.001, respectively). CONCLUSION: Our current study demonstrates the novel observation that a recently identified antimicrobial peptide, HNP 1-3, is expressed in neutrophils of acne inflammation but not in uninvolved skin of these patients. These results suggest that HNP 1-3 may contribute to the development of inflammatory lesions of acne. PMID- 19552719 TI - Oral manifestations of inflammatory bowel disease: a review based on the observation of six cases. AB - Inflammatory bowel disease (IBD) comprises two chronic, tissue-destructive, clinical entities: Crohn's disease (CD) and ulcerative colitis (UC), both immunologically based. Bowel symptoms are predominant, but extra-intestinal complications may occur, including involvement of the oral cavity. Oral involvement during IBD includes several types of lesions: the most common are aphthae; uncommon lesions include, among others, pyostomatitis vegetans and granulomatous lesions of CD. Starting with a presentation of six patients with oral manifestations, which were crucial for the final diagnosis of IBD, a review on the subject is presented. Oral involvement in IBD may be previous or simultaneous to the gastrointestinal symptoms. However, in the majority of cases, bowel disease precedes the onset of oral lesions by months or years. In many patients, the intestinal symptoms may be minimal and can go undetected; thus, most authors believe that the bowel must be thoroughly examined in all patients with suspected IBD even in the absence of specific symptoms. Usually, the clinical course of oral lesions is parallel to the activity of IBD; therefore, oral manifestations are a good cutaneous marker of IBD. PMID- 19552720 TI - Tinea nigra: a rare imported infection. PMID- 19552721 TI - Venous thromboembolism and ethnicity. AB - Venous thromboembolism (VTE) has long been considered a disease that affects predominantly white populations, a misconception resulting from a paucity of epidemiological data from non-Western countries, and the low incidence of hereditary thrombophilia in those of non-Caucasian background. Over the last decade, interest has grown in this area with the emergence of evidence that VTE is as prevalent, if not more so, in the black population and is also common in Asian groups. Much is still to be learned, as our current knowledge of hereditary thrombophilia and acquired risk factors do not fully explain the risk of VTE in non-Caucasian groups. This review summarises the current understanding of ethnic variation in VTE and highlights the need for further research in this area. PMID- 19552722 TI - Cytolytic T-cell response to the PASD1 cancer testis antigen in patients with diffuse large B-cell lymphoma. AB - The identification of immunogenic cancer testis antigens (CTAs) as immunotherapeutic targets represents one approach to improve treatment options for diffuse large B-cell lymphoma (DLBCL). We previously identified PASD1 [PAS (Per ARNT Sim) domain containing 1 (PASD1)], a DLBCL-associated CTA that was expressed in a range of hematopoietic malignancies. The aim of the present study was to investigate the presence of a cytotoxic T-cell (CTL) response to PASD1 in DLBCL patients. A significant gamma-interferon (IFN) release was detected in 21/29 HLA-A*0201-positive DLBCL patients (18 de novo DLBCL, two transformed DLBCL and one T-cell rich B-cell lymphoma) following short-term culture of their peripheral blood mononuclear cells stimulated with five HLA-A*0201-restricted PASD1 peptides. No significant responses were detected in 21 HLA-A*0201-negative DLBCL patients (12 de novo DLBCL, seven transformed DLBCL, two T-cell rich B-cell lymphoma) or six normal subjects. CTL cell lines were able to lyse PASD1-positive tumour cells in a major histocompatibility complex-Class I dependent manner. The presence of a gamma-IFN response correlated with PASD1 protein expression in the tumour cells in 12/15 cases studied. This is the first report of a CTL response to a CTA in DLBCL. Our results provide additional valuable evidence supporting PASD1 as a potential immunotherapeutic target for the treatment of DLBCL and other malignancies. PMID- 19552723 TI - Drug cross-resistance and therapy-induced resistance in chronic lymphocytic leukaemia by an enhanced method of individualised tumour response testing. AB - Previous results with individualised tumour response testing (ITRT) in vitro in chronic lymphocytic leukaemia (CLL) have consistently shown good correlation with patient response and survival. We describe here an improved test and report its use with samples from the Leukaemia Research Fund CLL4 randomised clinical trial and previously treated patients. ITRT was performed by the tumour response to anti-neoplastic compounds (TRAC) assay, a modification of the differential staining cytotoxicity (DiSC) assay. Improvements included drying drugs into wells before assay and using the Octospot system to cytocentrifuge eight spots of cells onto one microscope slide. We successfully tested 765/782 (98%) cellular blood samples received within 48 h of phlebotomy. Cross-resistance (Pearson's r > 0.7) in untreated CLL was found between similar drugs. Mitoxantrone (r = 0.31), cyclophosphamide (r = 0.35) and pentostatin (r = 0.29) had low cross-resistance with fludarabine. Treatment resulted in increased resistance to chlorambucil, cyclophosphamide, doxorubicin, mitoxantrone, corticosteroids, cladribine and fludarabine (P < 0.01) but not to pentostatin. These results provide further rationale for standard drug combinations such as fludarabine-mitoxantrone and fludarabine-mitoxantrone-cyclophosphamide and suggest possible pentostatin salvage in fludarabine-resistant patients. ITRT results could assist both in determining the best treatment for individual patients and in the design and rationale of future clinical trials. PMID- 19552724 TI - Association between a common IL10 distal promoter haplotype and IgE production in individuals with atopic dermatitis. AB - Atopic dermatitis (AD) is a genetically determined inflammatory skin disease characterized by abnormal cytokine production, including increased production of interleukin 10 (IL10). Single nucleotide polymorphisms (SNP) and haplotypes in the IL10 gene promoter region on chromosome 1q31-32 have been implicated in several inflammatory diseases, but generally, only SNPs proximal to the transcription start site (TSS) have been investigated. The aim of this study was to identify proximal, distal and combined haplotype sets in the IL10 promoter region and to study their association with clinical phenotypes in atopic dermatitis. SNPs at positions -3575, -2849, -2779, -2763, -1082, -851, -819 and 592 in the IL10 promoter region were genotyped in individuals with atopic dermatitis (n= 47) and nonatopic control subjects (n= 40) using polymerase chain reaction-based techniques and induced heteroduplex generator (IHG) analysis. Pan promoter, TSS-proximal and TSS-distal haplotypes were reconstructed using phase analysis. Fifteen haplotypes representing all eight SNPs were identified. Subgrouping identified four 4-locus and three 3-locus TSS-proximal haplotypes; and nine 4-locus TSS-distal haplotypes. No difference was found in haplotype or SNP frequencies between the AD and control groups, or between patients with mild or severe disease. However, a common 4-locus TSS-distal haplotype (TGAC) was significantly increased in patients with IgE levels over 1000 kIU L(-1). This study is the first to analyse the association between haplotype groups in the IL10 promoter region and clinical phenotypes in AD. We have demonstrated a significant association between the TSS-distal haplotype TGAC, and IgE levels in AD patients. It remains to be shown if there is an association between the TGAC haplotype and IL10 production, which might account for the stimulation of IgE production. PMID- 19552725 TI - Serotonin and GABA are colocalized in restricted groups of neurons in the larval sea lamprey brain: insights into the early evolution of neurotransmitter colocalization in vertebrates. AB - Colocalization of the classic neurotransmitters serotonin (5-HT) and gamma aminobutyric acid (GABA) (or the enzyme that synthesizes the latter, glutamate decarboxylase) has been reported in a few neurons of the rat raphe magnus obscurus nuclei. However, there are no data on the presence of neurochemically similar neurons in the brain of non-mammalian vertebrates. Lampreys are the oldest extant vertebrates and may provide important data on the phylogeny of neurochemical systems. The colocalization of 5-HT and GABA in neurons of the sea lamprey brain was studied using antibodies directed against 5-HT and GABA and confocal microscopy. Colocalization of the neurotransmitters was observed in the diencephalon and the isthmus. In the diencephalon, about 87% of the serotonergic cells of the rostral tier of the dorsal thalamus (close to the zona limitans) exhibited GABA immunoreactivity. In addition, occasional cells double-labelled for GABA and 5-HT were observed in the hypothalamic tuberal nucleus and the pretectum. Of the three serotonergic isthmic subgroups already recognized in the sea lamprey isthmus (dorsal, medial and ventral), such double-labelled cells were only observed in the ventral subgroup (about 61% of the serotonergic cells in the ventral subgroup exhibited GABA immunoreactivity). An equivalence between these lamprey isthmic cells and the serotonergic/GABAergic raphe cells of mammals is suggested. Present findings suggest that serotonergic/GABAergic neurons are more extensive in lampreys than in the rat and probably appeared before the separation of agnathans and gnathostomes. Cotransmission by release of 5-HT and GABA by the here-described lamprey brain neurons is proposed. PMID- 19552727 TI - The establishment of a new national network leads to quality improvement in childhood diabetes: implementation of the ISPAD Guidelines. AB - AIMS: To investigate whether implementation of International Society for Pediatric and Adolescent Diabetes (ISPAD) Guidelines and the establishment of a system for nationwide anonymous comparison, between treatment centres, of quality indicators for childhood diabetes could lead to improvement in diabetes care. METHODS: Children and adolescents with type-1 diabetes in Norway are treated at the public hospitals. Data were collected prospectively yearly according to standardized written instructions. Quality indicators were defined and benchmarked. HbA1c was measured at a central national Diabetes Control and Complications Trial (DCCT) standardized laboratory. RESULTS: The participation increased with 454 type-1 diabetes patients from eight clinics included in 2001 and 1658 patients from 25 clinics in 2005. The adherence rate in 2005 was 85% of all eligible patients from 25 of 26 pediatric clinics. The mean HbA1c of all clinics improved (8.6% in 2001 and 8.1% in 2005) and this was statistically significant (p < 0.01). The use of intensive insulin treatment increased from 56% to 78% (p < 0.01) and pumps from 8% to 37% (p < 0.01). The incidence of diabetes ketoacidosis (DKA) remained constant. The incidence of severe hypoglycemia declined insignificantly. The proportion of patients not screened yearly for microalbuminuria and retinopathy, according to ISPAD guidelines, decreased from 12% to 2% (p < 0.01) and from 42% to 27% (p < 0.01), respectively. All changes occurred gradually from 2001 to 2005. CONCLUSIONS: During the establishment of a system for benchmarking of diabetes treatment in Norway the outcomes showed significant improvements associated with changes in management and the quality of screening assessments. Benchmarking combined with organized quality meetings and discussions was effective to improve outcome on a national level. PMID- 19552726 TI - Vasoactive intestinal polypeptide immunoreactivity in the human cerebellum: qualitative and quantitative analyses. AB - Although autoradiographic, reverse transcription-polymerase chain reaction and immunohistochemical studies have demonstrated receptors for vasoactive intestinal polypeptide (VIP) in the cerebellum of various species, immunohistochemistry has never shown immunoreactivity for VIP within cerebellar neuronal bodies and processes. The present study aimed to ascertain whether VIP immunoreactivity really does exist in the human cerebellum by making a systematic analysis of samples removed post-mortem from all of the cerebellar lobes. The study was carried out using light microscopy immunohistochemical techniques based on a set of four different antibodies (three polyclonal and one monoclonal) against VIP, carefully selected on the basis of control tests performed on human colon. All of the antibodies used showed VIP-immunoreactive neuronal bodies and processes distributed in the cerebellar cortex and subjacent white matter of all of the cerebellum lobes, having similar qualitative patterns of distribution. Immunoreactive neurons included subpopulations of the main neuron types of the cortex. Statistical analysis of the quantitative data on the VIP immunoreactivity revealed by the different antibodies in the different cerebellar lobes did not demonstrate any significant differences. In conclusion, using four different anti VIP antibodies, the first evidence of VIP immunoreactivity is herein supplied in the human post-mortem cerebellum, with similar qualitative/quantitative patterns of distribution among the different cerebellum lobes. Owing to the function performed by VIP as a neurotransmitter/neuromodulator, it is a candidate for a role in intrinsic and extrinsic (projective) circuits of the cerebellum, in agreement with previous demonstrations of receptors for VIP in the cerebellar cortex and nuclei. As VIP signalling pathways are implicated in the regulation of cognitive and psychic functions, cerebral blood flow and metabolism, processes of histomorphogenesis, differentiation and outgrowth of nervous tissues, the results of this study could be applied to clinical neurology and psychiatry, opening new perspectives for the interpretation of neurodevelopment disorders and development of new therapeutic strategies in cerebellar diseases. PMID- 19552728 TI - Efficacy of ibandronate for the treatment of skeletal events in patients with metastatic breast cancer. AB - Patients with breast carcinoma often develop bone metastases that carry a high risk of complications. A randomized, placebo-controlled trial was conducted to evaluate the efficacy and safety of ibandronate in patients with metastatic bone disease following breast cancer. The primary efficacy end point of the study was the proportion of patients who developed skeletal-related events (SREs, defined as pathologic fracture, spinal cord compression, radiation therapy to bone, change in anti-neoplastic therapy and surgery to bone). Secondary end points included time to first skeletal event, skeletal morbidity rate (events/year) and time to progression of bone lesions. In 150 patients (148 [female symbol] / 2 [male symbol]) with breast carcinoma and bone metastases, treatment with intravenous ibandronate 6 mg over 15 min every 4 weeks for 24 months significantly reduced the proportion of patients who experienced an SRE compared with placebo (36% vs. 48%; P = 0.027). Time to first SRE was also delayed significantly (median 457 vs. 304 days; P = 0.007). Multiple event analysis showed that ibandronate reduced the risk of developing an SRE by 32% (hazard ratio = 0.69; 95% confidence interval 0.42-0.79; P = 0.003). In general, ibandronate was well tolerated with very rare grade 3 or 4 toxicity. In this study, ibandronate was shown to be significantly more effective than placebo as a treatment for metastatic bone disease from breast cancer using multiple end points. PMID- 19552729 TI - Social support and quality of life of prostate cancer patients after radiotherapy treatment. AB - Research suggests that social support can have an impact on health-related quality of life (HRQOL). Social support can be structural support (SSS) or functional support (FSS). Our study was designed to clarify the relationships between HRQOL, FSS and SSS. We conducted a cross-sectional survey and a detailed chart review. The study population was men attending a follow-up clinic after receiving radiotherapy for prostate cancer. Functional social support was measured by using the MOS Social Support Survey. Structural social support was measured by using questions adapted from the 1994-1995 National Population Health Survey conducted by Statistics Canada. Health-related quality of life was measured by using the European Organization for Research and Treatment of Cancer's QLQ-C30. We found a statistically significant positive correlation between FSS and HRQOL but no association between overall SSS and HRQOL. Worsening urinary symptoms were significantly associated with lower levels of FSS and with lower HRQOL. This study underscores that the perception of support (functional) is more important than the amount or size of support (structural). We also identified a subgroup of men who have lower FSS and lower HRQOL that suffer from urinary side effects of their treatment. Further research to clarify the relationship between FSS and urinary symptoms will also clarify how an intervention could improve the HRQOL of these men. PMID- 19552731 TI - Who visits mobile UK services providing cancer information and support in the community? AB - People can access a variety of sources of information and support when they have questions about cancer according to their needs. There are various sources of information and support for cancer beyond the health-care setting. In this study, we set out to assess reasons for visiting two mobile cancer information and support services in the UK during 2006. Data were collected about each visitor by staff on the mobile services. The two mobiles travelled to 109 UK locations over a 7-month period. Fifty-nine per cent of visitors were women. Thirty-one per cent of visitors had (had) cancer; very few were still undergoing treatment. For 95% of visitors the visit had been spontaneous rather than pre-planned, and 89% of visits lasted <15 min. Most visitors required information or support for themselves, but a third requested information for someone else. A quarter of enquiries were about cancer prevention and early detection (e.g. screening, genetic testing, lifestyle). The mobiles appear to serve an important function in providing information and support in the community where visitors can drop in for an informal conversation with trained members of staff to ask questions and receive support in relation to cancer. PMID- 19552730 TI - Mapping patients' experiences from initial change in health to cancer diagnosis: a qualitative exploration of patient and system factors mediating this process. AB - Delays in the diagnosis of cancer are common, and they are attributed to both patient and healthcare system factors. Minimizing such delays and improving early detection rates is a key goal of the new cancer reform strategy in England, in light of recent data showing that survival rates in the UK are low. The aim of this study was to explore the pathway from initial persistent change in health to diagnosis of cancer in a sample of patients from seven diagnostic groups in the UK and the factors mediating this process. Qualitative interviews with patients diagnosed with cancer were carried out. Seventy-five cancer patients discussed their pre-diagnosis experience as part of a broader exploration of their symptom experience for a larger study. Data were analysed by using content analysis and chart events. A broader range of mediating factors affecting and extending the patient pathway to diagnosis were reported in relation to lung, gastrointestinal and head and neck cancers and lymphoma, compared with breast, gynaecological and brain cancer patients. Many of the mediating factors were patient-related (e.g. misattribution of symptoms to common ailments, underestimation of the seriousness of the symptoms, self-medication or monitoring of symptoms, etc.). Primary care practitioner-factors were also prominent, including the exploration of firstly more common possibilities for treating the presenting symptoms without follow-up of persisting symptoms. Public health education about common cancer signs and symptoms, educational approaches in primary care to improve early diagnoses of cancer and updated guidelines for referral of suspected cancers should be enhanced before we can see any improvements in survival rates from cancer in the UK. PMID- 19552732 TI - Once-per-cycle pegfilgrastim in breast cancer patients treated with docetaxel/epidoxorubicin/cyclophosphamide. AB - The incidence of neutropenia following combination chemotherapy is significant in breast cancer and impairs patients' quality of life. Colony-stimulating factors significantly decrease the risk of febrile neutropenia (FN). Aim of the present study was to assess the efficacy and safety profile of once-per-cycle pegfilgrastim in reducing FN in breast cancer patients treated with docetaxel (75 mg/m(2)), epidoxorubicin (75 mg/m(2)), cyclophosphamide (500 mg/m(2)) administered every 3 weeks. Thirty-five breast cancer patients were enrolled. Chemotherapy was administered in adjuvant, neoadjuvant and metastatic setting respectively in 26, 4 and 5 patients. Toxicity was monitored with programmed clinical evaluation and blood sampling. All patients completed the therapeutic programme consisting of six cycles for overall 210 cycles. The FN appeared in 6 out of 35 patients (17%), requiring dose reduction in 3 patients. Hypertransaminasemia was registered in two patients. In one patient pegfilgrastim administration was stopped because of skin hypersensitivity reaction. In conclusion, pegfilgrastim was able to maintain doses and timing of docetaxel/epidoxorubicin/cyclophosphamide in almost all breast cancer patients treated in this series. The reduced need for daily administration of colony stimulating factors, blood sampling, antibiotic therapy and hospitalization has a significant impact in terms of both quality of life and pharmaco-economic evaluations. PMID- 19552733 TI - Highway driving performance and cognitive functioning the morning after bedtime and middle-of-the-night use of gaboxadol, zopiclone and zolpidem. AB - Gaboxadol is a selective extrasynaptic GABA(A) receptor agonist previously in development for the treatment of insomnia. Due to its short half-life (1.5-2 h) it is expected to be free from residual effects the next morning. The present study assessed the residual effects of evening and middle-of-the-night administration of 15 mg of gaboxadol on cognitive, psychomotor and driving performance. Twenty-eight healthy volunteers entered the study with 25 (12 women; mean age 31.4 years) completing a double-blind, placebo-controlled, active referenced five-way cross-over study. Each treatment night subjects ingested one capsule at 23:00 hours and one at 04:00 hours. Treatments were placebo at both times, 15 mg gaboxadol or 7.5 mg zopiclone followed by placebo, and placebo followed by 15 mg gaboxadol or 10 mg zolpidem. Effects on cognition and psychomotor performance were assessed between 07:30 and 08:30 hours and on driving between 09:00 and 10:00 hours. Driving, as measured by standard deviation of lateral position in an on-the-road driving test, was almost significantly (P < 0.07) impaired after evening administration of gaboxadol for the all-subjects completed set (n = 25) but significantly (P < 0.05) in the full analysis set (n = 28). Effects of all other active treatments on driving were significant. Evening administration of gaboxadol had minor effects on divided attention only, whereas middle-of-the-night administration impaired performance significantly in all tests except memory. Zolpidem and zopiclone impaired performance significantly in every test except tracking after zopiclone; 15 mg of gaboxadol can produce minor residual effects on driving after evening administration. Administration later at night is associated with moderately impairing residual effects on driving and psychomotor performance but not on memory. PMID- 19552734 TI - Prazosin modulates rapid eye movement sleep deprivation-induced changes in body temperature in rats. AB - Prolonged rapid eye movement sleep deprivation (REMSD) causes hypothermia and death; however, the effect of deprivation within 24 h and its mechanism(s) of action were unknown. Based on existing reports we argued that REMSD should, at least initially, induce hyperthermia and the death upon prolonged deprivation could be due to persistent hypothermia. We proposed that noradrenaline (NA), which modulates body temperature and is increased upon REMSD, may be involved in REMSD- associated body temperature changes. Adult male Wistar rats were REM sleep deprived for 6-9 days by the classical flower pot method; suitable free moving, large platform and recovery controls were carried out. The rectal temperature (Trec) was recorded every minute for 1 h, or once daily, or before and after i.p. injection of prazosin, an alpha-1 adrenergic antagonist. The Trec was indeed elevated within 24 h of REMSD which decreased steadily, despite continuation of deprivation. Prazosin injection into the deprived rats reduced the Trec within 30 min, and the duration of effect was comparable to its pharmacological half life. The findings have been explained on the basis of REMSD-induced elevated NA level, which has opposite actions on the peripheral and the central nervous systems. We propose that REMSD-associated immediate increase in Trec is due to increased Na-K ATPase as well as metabolic activities and peripheral vasoconstriction. However, upon prolonged deprivation, probably the persistent effect of NA on the central thermoregulatory sites induced sustained hypothermia, which if remained uncontrolled, results in death. Thus, our findings suggest that peripheral prazosin injection in REMSD would not bring the body temperature to normal, rather might become counterproductive. PMID- 19552735 TI - Evaluation of endothelial function using finger plethysmography. AB - BACKGROUND: The aims of this study were to establish the optimum duration of blood flow occlusion to obtain maximal response and to compare the response after lower-arm and upper-arm occlusion. METHODS: Pulse wave amplitude was analysed using a novel finger plethysmograph (EndoPat; Itamar). For measuring reactive hyperaemic index (RHI) induced by forearm cuff occlusion, 30 healthy subjects were examined at different days in a random order of four cuff occlusion times (1.5, 3, 5 and 8 min). RHI induced by 5 min upper-arm cuff occlusion was also measured in 20 subjects. RESULTS: Average RHI was lower with 1.5 and 3 min forearm occlusion compared with 8 min forearm occlusion (P = 0.002 and P = 0.024). There was no significant difference between values of 5 min and 8 min forearm occlusion and between 5 min forearm and 5 min upper-arm occlusion (P = 0.1). All subjects reported less discomfort after forearm occlusion compared with upper-arm occlusion. CONCLUSION: Maximum response was reached after 5 min of blood flow occlusion and therefore this occlusion time is recommended. The response after forearm and upper-arm occlusion did not differ significantly. Forearm occlusion might be preferred as this caused less discomfort. PMID- 19552737 TI - Sperm morphology and velocity are genetically codetermined in the zebra finch. AB - Sperm morphology (size and shape) and sperm velocity are both positively associated with fertilization success, and are expected to be under strong selection. Until recently, evidence for a link between sperm morphology and velocity was lacking, but recent comparative studies have shown that species with high levels of sperm competition have evolved long and fast sperm. It is therefore surprising that evidence for a phenotypic or genetic relationship between length and velocity within species is equivocal, even though sperm competition is played out in the intraspecific arena. Here, we first show that sperm velocity is positively phenotypically correlated with measures of sperm length in the zebra finch Taeniopygia guttata. Second, by using the quantitative genetic "animal model" on a dataset from a multigenerational-pedigreed population, we show that sperm velocity is heritable, and positively genetically correlated to a number of heritable components of sperm length. Therefore, selection for faster sperm will simultaneously lead to the evolution of longer sperm (and vice versa). Our results provide, for the first time, a clear phenotypic and genetic link between sperm length and velocity, which has broad implications for understanding how recently described macroevolutionary patterns in sperm traits have evolved. PMID- 19552738 TI - The diversity and evolution of batesian mimicry in Papilio swallowtail butterflies. AB - Papilio swallowtail butterflies exhibit a remarkable diversity of Batesian mimicry, manifested in several sex-limited and polymorphic types. There is little understanding of how this diversity is distributed within Papilio, and how different mimicry types have evolved in relation to each other. To answer these questions, I present a graphical model that connects various mimicry types by hypothetical character state changes within a phylogenetic framework. A maximum likelihood analysis of evolution of mimicry types on the Papilio phylogeny showed that sexually monomorphic mimicry and female-limited mimicry have evolved repeatedly but predominantly independently in different clades. However, transitions between these mimicry types are rarely observed. The frequency distribution of character state changes was skewed in favor of the evolution of mimicry, whereas many theoretically plausible character state changes, especially evolutionary loss of mimicry, were not evident. I discuss these findings in relation to studying the tempo of evolutionary change, loss of traits, and directionality and connectivity among character states. The pathway approach and phylogenetic patterns of mimicry demonstrated in Papilio are useful to test novel hypotheses regarding the diversity and evolutionary directionality of Batesian mimicry in other systems. PMID- 19552739 TI - Sexual selection is involved in speciation in a land snail radiation on crete. AB - We investigated the importance of sexual selection in facilitating speciation in a land snail radiation on Crete. We used differences in the genitalia of the Cretan Xerocrassa species as potential indices of sexual selection. First, we rejected the hypothesis that differences in the genitalia of the Xerocrassa species can be explained by genetic drift using coalescent simulations based on a mitochondrial gene tree. Second, we showed that there is no evidence for the hypothesis that the differences in the genitalia can be explained by natural selection against hybrids under the assumption that this is more likely in geographically overlapping species pairs and clades. Third, we showed that there is a positive scaling between male spermatophore-producing organs and female spermatophore-receiving organs indicating sexual coevolution. The spermatophore enables the sperm to escape from the female gametolytic organ. Thus, the coevolution might be a consequence of sexual conflict or cryptic female choice. Finally, we showed that the evolution of differences in the length of the flagellum that forms the tail of the spermatophore is concentrated toward the tips of the tree indicating that it is involved in speciation. If speciation is facilitated by sexual selection, niches may remain conserved and nonadaptive radiation may result. PMID- 19552740 TI - Effect of recombinant human erythropoietin on insulin resistance in hemodialysis patients. AB - Insulin resistance is a characteristic feature of uremia. Insulin resistance and concomitant hyperinsulinemia are present irrespective of the type of renal disease. Treatment with recombinant human erythropoietin (rHuEPO) was said to be associated with improvement in insulin sensitivity in uremic patients. The aim of this study was to compare insulin resistance in adult uremic hemodialysis (HD) patients including diabetic patients treated with or without rHuEPO. A total of 59 HD patients were studied, patients were divided into 2 groups of subjects: 30 HD patients on regular rHuEPO treatment (group A), and 29 HD patients not receiving rHuEPO (group B) diabetic patients were not excluded. Full medical history and clinical examination, hematological parameters, lipid profile, serum albumin, parathyroid horomone, Kt/V, fasting glucose, and insulin levels were measured in all subjects. Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) was used to compare insulin resistance. The results of this study showed that the mean insulin level of HD patients treated with rHuEPO (group A) (17.5 +/- 10.6 microU/mL) was significantly lower than patients without rHuEPO (group B) (28.8 +/- 7.7 microU/mL), (P<0.001). Homeostasis Model Assessment of Insulin Resistance levels in group A were significantly lower than in group B (3.8 +/- 2.97, 7.98 +/- 4.9, respectively, P<0.001). Insulin resistance reflected by HOMA-IR levels among diabetic patients in group A was significantly lower than among diabetic patients in group B (3.9 +/- 3.2, 9.4 +/- 7.2, respectively, P<0.001). Also, HOMA-IR levels among nondiabetic patients in group A were significantly lower than among nondiabetic patients in group B (3.7 +/- 2.85, 6.9 +/- 1.43, respectively, P<0.01). We found a statistically significant negative correlation between duration of erythropoietin treatment, fasting blood glucose, insulin levels, and insulin resistance (r=-0.62, -0.71, and -0.57, P<0.001). Patients treated with rHuEPO showed less insulin resistance compared with patients not treated with rHuEPO in diabetic and nondiabetic patients and, duration of erythropoietin treatment is negatively correlated with insulin levels and insulin resistance in HD patients. PMID- 19552741 TI - Medication changes based on echocardiography in dialysis patients. AB - Kidney/Disease Outcome Quality Initiative (K/DOQI) guidelines recommend baseline echocardiography at the initiation of dialysis and every 3 years thereafter in patients for early detection of cardiac disease to optimize medical therapy. Because dialysis patients are at increased cardiovascular risk and thus most are already on cardioprotective medications, we hypothesize that serial screening echocardiography will not alter cardioprotective medications in dialysis patients. Retrospective analysis of medication administration of 231 dialysis patients was conducted. Patients were divided into 2 groups, those with and those without echocardiograms. Medication changes post echocardiography were compared with subjects without echocardiograms at comparable time points. The primary end point was the number of medication class changes that occurred in 2 months post echocardiography. Medication classes examined were beta blockers (BB), angiotensin converting enzyme inhibitors or angiotensin receptor blockers (ACEI/ARB), nitrates, calcium channel blockers (CCB), and statins. In the Echo group, there were 29 (19%) subjects with at least 1 medication class change post echocardiography, compared with 121 (81%) subjects without change. The number of patients on specific medication classes before and after echocardiography were BB (90 [60%] vs. 97 [65%], P=0.05), ACEI/ARB (74 [49%] vs. 82 [55%], P=0.01), nitrates (34 [23%] vs. 33 [22%], P=0.56), CCB (77 [51%] vs. 79 [53%], P=0.56), and statins (69 [46%] vs. 70 [47%], P=0.71). When compared with the No Echo group, there was no significant change in number of any medication classes. The occurrence of medication changes post echocardiography in dialysis patients is low and is not different than changes in routine care of dialysis patients without echocardiograms. Thus, serial screening echocardiography may not have added benefit to optimizing medical management of cardiovascular disease in dialysis patients. Further studies are warranted to demonstrate evidence for the use of serial screening echocardiography in this high-risk population. PMID- 19552742 TI - The impact of clinical trials legislation on clinical pharmacology: problems and solutions. PMID- 19552743 TI - Orphan drug development is not taking off. PMID- 19552744 TI - Pharmacokinetic- pharmacodynamic analysis of the role of CYP2C19 genotypes in short-term rabeprazole-based triple therapy against Helicobacter pylori. AB - AIMS: The aim was to explore the role of CYP2C19 polymorphism in short-term rabeprazole-based triple therapy against Helicobacter pylori infection. METHODS: Patients with H. pylori infection were tested for CYP2C19 genotype as poor metabolizers (PMs) or extensive metabolizers (EMs, homozygous EM or heterozygous EM) and given rabeprazole for 7 days. Antibiotics (clarithromycin and amoxicillin) were given on days 1-4, days 4-7, or days 1-7. A direct link model with an effect compartment was used in the population pharmacokinetic pharmacodynamic analysis. The status of H. pylori infection was evaluated. RESULTS: Rabeprazole clearance was lower in CYP2C19 PMs than in EMs (with average values of 10.7 vs. 16.8 l h(-1) in PMs and EMs, respectively), resulting in higher plasma levels in the former group. The values of EC(50) and k(eo) of gastrin response increased with multiple doses of rabeprazole. The k(eo) values were lower in CYP2C19 PMs than in EMs on day 1 (0.012 vs. 0.017 x 10(-4) l min( 1)), and higher than in EMs on day 4 (0.804 vs. 0.169 x 10(-4) l min(-1)) of rabeprazole treatment. The predicted gastrin-time profile showed a higher response in CYP2C19 PMs than in EMs on days 4 and 7. Helicobacter pylori was eradicated in all CYP2C19 PMs except in one patient infected by a resistant strain. In contrast, in CYP2C19 EMs the eradication rates ranged from 58 to 85%. CONCLUSIONS: CYP2C19 genotypes play a role in H. pylori eradication therapy. Rabeprazole-based short-term triple therapy may be applicable in CYP2C19 PMs for H. pylori eradication. PMID- 19552745 TI - Amiodarone concentrations in plasma and fat tissue during chronic treatment and related toxicity. AB - AIMS: To determine if amiodarone, highly lipophilic, accumulates in excess with respect to dose in fat tissue during long-term administration, and study if plasma and fat tissue concentrations are correlated with adverse effects. METHODS: Trough concentrations of amiodarone and N-desethyl-amiodarone were measured simultaneously in plasma and fat tissue, in 30 consecutive patients treated with amiodarone for 3 months to 12 years. Subcutaneous adipose tissue was obtained by needle aspiration from lumbar and abdominal areas. Concentrations were measured by liquid chromatography-tandem mass spectrometry. RESULTS: Plasma levels of amiodarone and N-desethyl-amiodarone were significantly correlated with daily maintenance doses (R= 0.52, P= 0.003). Amiodarone concentrations in fat tissue were four to 226 times (mean 55) higher than in plasma, and well correlated with plasma levels (R= 0.68, P < 0.001). Concentrations of amiodarone and N-desethyl-amiodarone in adipose tissue did not significantly increase with higher total cumulated doses or longer treatment duration. Nine of 12 patients who had received amiodarone for > or =2 years developed clinically important adverse effects, predominantly hypothyroidism (n= 6), compared with two of 18 patients treated for less time (relative risk 6.75; 95% confidence interval 1.8, 26). The incidence of those adverse effects was not significantly associated with amiodarone concentrations, whether in plasma or in adipose tissue. CONCLUSIONS: We found no evidence of excessive or unexpected accumulation of amiodarone in fat tissue on long-term administration. Late amiodarone adverse effects, particularly hypothyroidism, are associated with longer exposure times, but do not seem to be explained by higher concentrations in plasma or in fat tissue. PMID- 19552746 TI - Assessment of a pharmacokinetic and pharmacodynamic interaction between simvastatin and anacetrapib, a potent cholesteryl ester transfer protein (CETP) inhibitor, in healthy subjects. AB - AIMS: Anacetrapib is an orally active, potent inhibitor of cholesteryl ester transfer protein (CETP), which is in development for the treatment of dyslipidaemia. Because of the likely use of anacetrapib with hydroxy-3 methylglutaryl coenzyme A reductase inhibitors, we aimed to evaluate the potential for a pharmacokinetic interaction with simvastatin. METHODS: A randomized, two-period, two-treatment, balanced, open-label, crossover study in 12 healthy subjects was performed. Subjects received simvastatin 40 mg alone or anacetrapib 150 mg co-administered with simvastatin 40 mg, once daily. Both treatments were administered following a low-fat breakfast for 14 days, separated by a wash-out period of at least 14 days. Safety and tolerability, simvastatin and simvastatin acid concentrations, and lipoproteins, were assessed. RESULTS: Both treatments were well tolerated. The pharmacokinetics of simvastatin and simvastatin acid were similar with and without anacetrapib administration {AUC(0 24 h) geometric mean ratio [90% confidence interval (CI)] for simvastatin acid and simvastatin were 1.36 [1.17, 1.57] and 1.30 [1.14, 1.47], respectively} based on the prespecified comparability bounds of (0.50, 2.00). Treatment with simvastatin alone led to a mean (95% CI) % reduction from baseline in low-density lipoprotein-cholesterol (LDL-C) of -36% (-27, -46) compared with a reduction of 54% (-44, -63) for anacetrapib co-administered with simvastatin. CONCLUSIONS: There appears to be no clinically meaningful effect of anacetrapib on the pharmacokinetic parameters of simvastatin. When co-administered with simvastatin, anacetrapib appeared to exhibit incremental LDL-C-lowering efficacy, due to CETP inhibition. Co-administration of anacetrapib and simvastatin was well tolerated. PMID- 19552747 TI - The pharmacokinetics and pharmacodynamics of warfarin in combination with ambrisentan in healthy volunteers. AB - AIMS: Ambrisentan is an oral, propanoic acid-based endothelin receptor antagonist often co-administered with warfarin to patients with pulmonary arterial hypertension. The aim of this study was to evaluate the potential for ambrisentan to affect warfarin pharmacokinetics and pharmacodynamics. METHODS: In this open label cross-over study, 22 healthy subjects received a single dose of racemic warfarin 25 mg alone and after 8 days of ambrisentan 10 mg once daily. Assessments included exposure (AUC(0-last)) and maximum plasma concentration (C(max)) for R- and S-warfarin, and International Normalized Ratio maximum observed value (INR(max)) and area under the curve (INR(AUC(0-last))). The effects of warfarin on ambrisentan steady-state pharmacokinetics and the safety of ambrisentan/warfarin co-administration were assessed. Data are presented as geometric mean ratios. RESULTS: Ambrisentan had no significant effects on the AUC(0-last) of R-warfarin [104.7; 90% confidence interval (CI) 101.7, 107.7) or S warfarin (101.6; 90% CI 98.4, 105.0). Similarly, ambrisentan had no significant effects on the C(max) of R-warfarin (91.6; 90% CI 86.2, 97.4) or S-warfarin (89.9; 90% CI 84.8, 95.3). Consistent with these observations, little pharmacodynamic change was observed for INR(max) (85.3; 90% CI 82.4, 88.2) or INR(AUC(0-last)) (93.0; 90% CI 90.8, 95.3). In addition, co-administration of warfarin did not alter ambrisentan steady-state pharmacokinetics. Adverse events were infrequent, and there were no bleeding adverse events. CONCLUSIONS: Multiple doses of ambrisentan had no clinically relevant effects on the pharmacokinetics and pharmacodynamics of a single dose of warfarin. Therefore, significant dose adjustments of either drug are unlikely to be required with co-administration. PMID- 19552748 TI - Enantioselective disposition of fexofenadine with the P-glycoprotein inhibitor verapamil. AB - AIMS: The aim was to compare possible effects of verapamil, as a P-glycoprotein (P-gp) inhibitor, on the pharmacokinetics of each fexofenadine enantiomer, as a P gp substrate. METHODS: Thirteen healthy Japanese volunteers (10 male and three female) were enrolled. In a randomized, two-phase, crossover design, verapamil was dosed 80 mg three times daily (with total daily doses of 240 mg) for 6 days, and on day 6, a single 120-mg dose of fexofenadine was administered along with an 80-mg dose of verapamil. Subsequently, fexofenadine was administered alone after a 2-week wash-out period. The plasma concentrations of fexofenadine enantiomers were measured up to 24 h after dosing. RESULTS: During the control phase, the mean AUC(0-infinity) of S(-)- and R(+)-fexofenadine was 700 ng h(-1) ml(-1)[95% confidence interval (CI) 577, 823] and 1202 ng h(-1) ml(-1) (95% CI 1007, 1396), respectively, with a significant difference (P < 0.001). Verapamil had a greater effect on the pharmacokinetic parameters of S(-)-fexofenadine compared with those of the R(+)-enantiomer, and increased AUC(0-infinity) of S(-)-fexofenadine and R(+)-fexofenadine by 3.5-fold (95% CI of differences 1.9, 5.1; P < 0.001) and by 2.2-fold (95% CI of differences 1.7, 3.0; P < 0.001), respectively. The R/S ratio for the AUC(0-infinity) was reduced from 1.76 to 1.32 (P < 0.001) by verapamil treatments. CONCLUSION: This study indicates that P-gp plays a key role in the stereoselectivity of fexofenadine pharmacokinetics, since the pharmacokinetics of fexofenadine enantiomers were altered by the P-gp inhibitor verapamil, and this effect was greater for S-fexofenadine compared with R-fexofenadine. PMID- 19552749 TI - Effects of Schisandra sphenanthera extract on the pharmacokinetics of midazolam in healthy volunteers. AB - AIMS: To assess the effect of Schisandra sphenanthera extract (SchE) on the pharmacokinetics of midazolam, a probe drug of CYP3A, and its metabolite 1' hydroxy midazolam in healthy volunteers. METHODS: Twelve healthy male volunteers were orally treated with SchE, three capsules twice daily for 7 days. Pharmacokinetic investigations of oral midazolam administration at 15 mg were performed both before and at the end of the SchE treatment period. The plasma midazolam and 1'-hydroxy midazolam concentrations were determined by high performance liquid chromatography-tandem mass spectrometry. Estimated pharmacokinetic parameters before and with SchE were calculated with noncompartmental techniques. RESULTS: Following administration of SchE, the average increases (%) of individual increases in AUC, AUMC and C(max) of midazolam were 119.4% [95% confidence interval (CI) 83.9, 155.0], 183.4% (95% CI 120.5, 246.2) and 85.6% (95% CI 14.4, 156.9), respectively (P < 0.01 or 0.05). On average, there was a 133.3% (95% CI 8.9, 257.7) increase in midazolam t(max) (P < 0.01). The average decrease (%) in CL/F was 52.1% (95% CI 44.9, 59.4) (P < 0.01). No significant changes were seen in midazolam half-life. After co-administration of SchE, the average increase (%) in t(max) of 1'-hydroxy midazolam was 150.0% (95% CI 22.2, 277.8) (P < 0.05). No significant differences were observed in the other pharmacokinetic parameters of 1'-hydroxy midazolam. CONCLUSIONS: SchE can markedly increase the oral bioavailability of midazolam in healthy volunteers. SchE is an inhibitor of CYP3A and has a high susceptibility to alter the disposition of drugs metabolized by CYP3A. PMID- 19552751 TI - Comparing patients' and healthcare professionals' ADR reports after media attention: the broadcast of a Dutch television programme about the benefits and risks of statins as an example. AB - AIMS: To compare adverse drug reaction (ADR) reports from patients and health professionals after the broadcast of a Dutch television consumer programme about the benefits and risks of statins. METHODS: We performed a quantitative and qualitative analysis on patients' and health professionals' reports of ADRs to statins. These reports were received by the Netherlands Pharmacovigilance Centre Lareb between March 2007 and August 2007. Quantitative data consisted of patient age and gender, number of received reports and characteristics of the report (most frequently reported ADRs, seriousness, drug discontinuation and outcome of the reported reaction). Open text fields in the ADR reporting form were categorized and a content analysis was carried out. RESULTS: Media attention led to a peak in patient reporting of ADRs but not in reporting by health professionals. There were no differences between patient and health professional reports in seriousness of the ADRs and drug cessation. Patients reported nonrecovery more often than health professionals. The TV programme is mentioned as a reason for drug discontinuation in almost 30 reports. Patients often felt that they did not receive sufficient information and that their concerns were not adequately addressed by healthcare professionals. CONCLUSIONS: Media attention affects drug use and ADR reporting by patients. Patient reports can provide additional information, making them a useful source of information next to health professional reports. Content analysis provides vital insights into the impact of statins on daily life, and patients' concerns about adverse reactions should be recognized in reports to national pharmacovigilance centres. PMID- 19552750 TI - Influence of pharmaceutical care on health outcomes in patients with Type 2 diabetes mellitus. AB - AIMS: To examine the influence of a pharmaceutical care programme on disease control and health-related quality of life in Type 2 diabetes patients in the United Arab Emirates. METHODS: A total of 240 Type 2 diabetes patients were recruited into a randomized, controlled, prospective clinical trial with a 12 month follow-up. A range of clinical measures, medication adherence and health related quality of life (Short Form 36) were evaluated at baseline and up to 12 months. Intervention group patients received pharmaceutical care from a clinical pharmacist, whereas control group patients received their usual care from medical and nursing staff. The primary outcome measure was change in HbA(1c). British National Formulary and Framingham scoring methods were used to estimate changes in 10-year coronary heart disease risk scores in all patients. RESULTS: A total of 234 patients completed the study. Significant reductions (P < 0.001) in mean values (baseline vs. 12 months; 95% confidence interval) of HbA(1c)[8.5% (8.3, 8.7) vs. 6.9% (6.7, 7.1)], systolic [131.4 mmHg (128.1, 134.7) vs. 127.2 mmHg (124.4, 130.1)] and diastolic blood pressure [85.2 mmHg (83.5, 86.8) vs. 76.3 mmHg (74.9, 77.7)] were observed in the intervention group; no significant changes were noted in the control group. The mean Framingham risk prediction score in the intervention group was 10.56% (9.7, 11.4) at baseline; this decreased to 7.7% (6.9, 8.5) (P < 0.001) at 12 months but remained unchanged in the control group. CONCLUSIONS: The pharmaceutical care programme resulted in better glycaemic control and reduced cardiovascular risk scores in Type 2 diabetes patients over a 12-month period. PMID- 19552752 TI - Determinants of the drug utilization profile in the paediatric population in Italy's Lombardy Region. AB - AIMS: To evaluate the intraregional differences in drug prescribing to children and adolescents. METHODS: Prescriptions reimbursed by the National Health System, involving 1543 203 children and adolescents <18 years old and dispensed during 2005 by the retail pharmacies of 15 local health units (LHU) in the Lombardy Region, were analysed. Logistic regression analysis was performed to evaluate the association between drug prescription and age, gender, prescriber, and setting. RESULTS: A total of 747 790 youths (48%) received at least one drug prescription. The prescription prevalence rate was highest in children 1-5 years old (65%), decreased with increasing age to 38% in adolescents, and was slightly higher in boys than in girls. Antibiotics and anti-asthmatics were the most prescribed therapeutic classes. Amoxicillin + clavulanic acid was the most prescribed drug (18% of children; 20% of packages). Large differences were found in the drug prescription prevalence rate between the different LHUs. The rate ranged between 38.4 and 54.8%, and was not correlated to hospitalization rate in the paediatric population. Being 1-5 years old [odds ratio (OR) 4.51, 95% confidence interval (CI) 4.43, 4.58] and living in the eastern part of the region (OR 2.06, 95% CI 1.99, 2.13) were the factors associated with the highest risk of drug exposure. CONCLUSIONS: The results resemble the profiles observed in other Italian contexts, in particular concerning the wide use of antibiotics and anti asthmatics. However, large differences were found between LHUs, highlighting the need for more detailed investigations on therapeutic needs, drug use, and related variables in different geographic contexts. PMID- 19552753 TI - Electrocardiogram changes and arrhythmias in venlafaxine overdose. AB - AIMS: To investigate serial electrocardiogram (ECG) parameters, haemodynamic changes and arrhythmias following venlafaxine overdose. METHODS: The study included 369 venlafaxine overdoses in 273 patients presenting to a toxicology unit where an ECG was available. Demographic information, details of ingestion, haemodynamic effects [heart rate and blood pressure (BP)] and complications (arrhythmias and conduction defects) were obtained. ECG parameters (QT, QRS) were measured manually and analysed by visual inspection, including plotting QT-HR pairs on a QT nomogram. RESULTS: The median ingested dose was 1500 mg [interquartile range (IQR) 600-3000 mg; range 75-13 500 mg). Tachycardia occurred in 54% and mild hypertension (systolic BP >140 mmHg) in 40%. Severe hypertension (systolic BP >180 mmHg) and hypotension (systolic BP <90 mmHg) occurred in 3% and 5%, respectively. No arrhythmias occurred based on continuous telemetry, and conduction defects were found in only seven of 369 admissions; five of these conduction defects were pre-existing abnormalities. In 22 admissions [6%, 95% confidence interval (CI) 4-10] there was an abnormal QT-HR pair, with larger doses being more likely to be associated with an abnormal QT. The median maximum QRS width was 85 ms (IQR 80-90 ms; range 70-145 ms) and the QRS was greater than 120 ms in only 24 admissions (7%, 95% CI 4-10). CONCLUSIONS: Venlafaxine overdose causes only minor abnormalities in the QT and QRS intervals, unlikely to be associated with major arrhythmias, except possibly with large doses. PMID- 19552754 TI - Cetirizine-induced anaphylaxis: a rare adverse drug reaction. PMID- 19552755 TI - Acute oxcarbazepine overdose in an autistic boy. PMID- 19552756 TI - Genetic variants in the Runt-related transcription factor 3 gene contribute to gastric cancer risk in a Chinese population. AB - Runt-related transcription factor 3 (RUNX3) is a well known gene for its functions in gastric cancer suppression, but the effect of its genetic variations on the risk of gastric cancer remains unclear. In this study, ten tagging single nucleotide polymorphisms (tSNPs) of the RUNX3 gene were selected and genotyped in a hospital-based case-control study of 312 gastric cancer patients and 329 cancer free controls in a Chinese population. In the single-locus analysis, three RUNX3 intronic tSNPs associated with significantly increased risk of gastric cancer were observed: the SNP3 rs11249206 CC genotype (adjusted odds ratio [OR] = 1.75, 95% confidence interval [CI] = 1.03-2.99), compared with the TT genotype; the SNP7 rs760805 AA genotype (adjusted OR = 1.82, 95% CI = 1.14-2.92), compared with the TT genotype; and the SNP8 rs2236852 GG genotype (adjusted OR = 1.69, 95% CI = 1.05-2.72), compared with the AA genotype. In the combined analyses of these three tSNPs, we found that the combined genotypes with four to six variant (risk) alleles (i.e. SNP3 C, SNP7 A, and SNP8 G alleles) were associated with an increased risk of gastric cancer compared with those with one to three variant (risk) alleles (adjusted OR = 2.00, 95% CI = 1.41-2.85), and this increased risk was more pronounced among subgroups of age > or =65 years, never smokers, and never drinkers. However, no significant association was observed in the clinicopathological features analyses. In conclusion, the RUNX3 genetic variants may modulate the risk of gastric cancer in a Chinese population. Further larger and functional studies are warranted to validate the findings. PMID- 19552757 TI - Overexpression of AIB1 predicts resistance to chemoradiotherapy and poor prognosis in patients with primary esophageal squamous cell carcinoma. AB - AIB1 (amplified in breast cancer 1) is frequently overexpressed in esophageal squamous cell carcinoma (ESCC), but the significance of AIB1 expression in chemoradiotherapy (CRT) sensitivity and its effect on prognosis are still unclear. In this study, the expression of AIB1 was examined by immunohistochemistry in 98 biopsy specimens of primary ESCC patients treated with definitive CRT. AIB1 overexpression was found in 63/98 (64.3%) of the ESCCs. There was a significant association between AIB1 overexpression and distant lymph node metastases (P = 0.011), but not regional lymph node metastases. In the M0 subgroup, overexpression of AIB1 was observed more frequently in stage T4 than in stage T2-3 (66.7%vs 38.5%, P = 0.031). In addition, AIB1 expression was the only factor that showed a significant correlation with CRT response, in which overexpression of AIB1 was observed more frequently in the CRT resistant group than in the CRT effective group (86.5%vs 50.8%, P < 0.001). Univariate analysis revealed that AIB1 overexpression was associated with poor progression-free survival (PFS) (P < 0.001) and poor disease-specific survival (DSS) (P <0.001). Furthermore, AIB1 expression could stratify patient survival in stages T2-3, T4, N1, and M0 (P < 0.05), as well as in the CRT effective group (P < 0.05), and AIB1 overexpression and CRT resistance were evaluated as significant independent prognostic factors for both PFS and DSS in multivariate analysis. These findings suggest that overexpression of AIB1 is a useful predictor of CRT resistance and an independent molecular marker of poor prognosis for ESCC patients. PMID- 19552758 TI - Melatonin improves placental efficiency and birth weight and increases the placental expression of antioxidant enzymes in undernourished pregnancy. AB - Melatonin participates in circadian, seasonal and reproductive physiology. Melatonin also acts as a potent endogenous antioxidant by scavenging free radicals and upregulating antioxidant pathways. The placenta expresses melatonin receptors and melatonin protects against oxidative damage induced in rat placenta by ischemia-reperfusion. One of the most common complications in pregnancy is a reduction in fetal nutrient delivery, which is known to promote oxidative stress. However, whether melatonin protects placental function and fetal development in undernourished pregnancy is unknown. Here, we investigated the effects of maternal treatment with melatonin on placental efficiency, fetal growth, birth weight and protein expression of placental oxidative stress markers in undernourished pregnancy. On day 15 of pregnancy, rats were divided into control and undernourished pregnancy (35% reduction in food intake), with and without melatonin treatment (5 microg/mL drinking water). On day 20 of gestation, fetal biometry was carried out, the placenta was weighed and subsequently analyzed by Western blot for xanthine oxidase, heat shock protein (HSP) 27 and 70, catalase, manganese superoxide dismutase (Mn-SOD) and glutathione peroxidase 1 (GPx-1). A separate cohort was allowed to deliver to assess effects on birth weight. Maternal undernutrition led to a fall in placental efficiency, disproportionate intrauterine growth retardation and a reduction in birth weight. Maternal treatment with melatonin in undernourished pregnancy improved placental efficiency and restored birth weight, and it increased the expression of placental Mn-SOD and catalase. The data show that in pregnancy complicated by undernutrition, melatonin may improve placental efficiency and birth weight by upregulating placental antioxidant enzymes. PMID- 19552759 TI - Melatonin protects kidney grafts from ischemia/reperfusion injury through inhibition of NF-kB and apoptosis after experimental kidney transplantation. AB - Free radicals are involved in pathophysiology of ischemia/reperfusion injury (IRI). Melatonin is a potent scavenger of reactive oxygen and nitrogen species. Thus, this study was designed to elucidate its effects in a model of rat kidney transplantation. Twenty Lewis rats were randomly divided into 2 groups (n = 10 animals each). Melatonin (50 mg/kg BW) dissolved in 5 mL milk was given to one group via gavage 2 hr before left donor nephrectomy. Controls were given the same volume of milk only. Kidney grafts were then transplanted into bilaterally nephrectomized syngeneic recipients after 24 hr of cold storage in Histidine Tryptophan-Ketoglutarate solution. Both graft function and injury were assessed after transplantation through serum levels of blood urea nitrogen (BUN), creatinine, transaminases, and lactate dehydrogenase (LDH). Biopsies were taken to evaluate tubular damage, the enzymatic activity of superoxide dismutase (SOD) and lipid hydroperoxide (LPO), and the expression of NF-kBp65, inducible nitric oxide synthase (iNOS), caspase-3 as indices of oxidative stress, necrosis, and apoptosis, respectively. Melatonin improved survival (P < 0.01) while decreasing BUN, creatinine, transaminases, and LDH values up to 39-71% (P < 0.05). Melatonin significantly reduced the histological index for tubular damage, induced tissue enzymatic activity of SOD while reducing LPO. At the same time, melatonin down regulated the expression of NF-kBp65, iNOS, and caspase-3. In conclusion, donor preconditioning with melatonin protected kidney donor grafts from IRI-induced renal dysfunction and tubular injury most likely through its anti-oxidative, anti apoptotic and NF-kB inhibitory capacity. PMID- 19552760 TI - Cloning and characterization of a Chlamydomonas reinhardtii cDNA arylalkylamine N acetyltransferase and its use in the genetic engineering of melatonin content in the Micro-Tom tomato. AB - Melatonin is found in a wide variety of plant species. Several investigators have studied the physiological roles of melatonin in plants. However, its role is not well understood because of the limited information on its biosynthetic pathway. To clarify melatonin biosynthesis in plants, we isolated a cDNA-coded arylalkylamine N-acetyltransferase (AANAT), a possible limiting enzyme for melatonin biosynthesis, from Chlamydomonas reinhardtii (designated as CrAANAT). The predicted amino acid sequence of CrAANAT shares 39.0% homology to AANAT from Ostreococcus tauri and lacks cAMP-dependent protein kinase phosphorylation sites in the N- and C-terminal regions that are conserved in vertebrates. The enzyme activity of CrAANAT was confirmed by in vitro assay using Escherichia coli. Transgenic plants constitutively expressing the CrAANAT were produced using Micro Tom, a model cultivar of tomato (Solanum lycopersicum L.). The transgenic Micro Tom exhibited higher melatonin content compared with wild type, suggesting that melatonin was synthesized from serotonin via N-acetylserotonin in plants. Moreover, the melatonin-rich transgenic Micro-Tom can be used to elucidate the role of melatonin in plant development. PMID- 19552761 TI - Exogenous melatonin positively influences follicular dynamics, oocyte developmental competence and blastocyst output in a goat model. AB - The role of melatonin in modulating mammalian reproduction is of particular interest; however, its effects on ovarian follicles and their oocytes still remain to be characterized. This study determined the influence of melatonin treatment on follicular growth patterns and on in vitro oocyte developmental competence. In a first experiment, the effects of melatonin supplementation on follicular dynamics were evaluated using daily transrectal ultrasonographies for 21 days, in 7 multiparous Sarda goats receiving a subcutaneous implant of 18 mg of melatonin and in 5 control untreated does. Melatonin caused more follicular waves (5.2 +/- 0.2 versus 4 +/- 0.3; P < 0.05) as the waves were shortened at around 2 days when compared with the non-melatonin treated control goats (P < 0.001). Oocyte developmental competence was evaluated in a second experiment by applying procedures for in vitro embryo production. There were no significant differences in the total number of oocytes obtained from 6 control (n = 192) and 7 melatonin-treated (n = 265) goats given follicle stimulating hormone to induce follicular development. Differences in oocyte developmental competence between the two groups became evident after in vitro fertilization and culture; melatonin increased the rate of cleaved oocytes in comparison with control animals (82.5 versus 63.4%; P < 0.001), advanced timing of embryo development and enhanced blastocyst output (31.5 versus 16.3%; P < 0.01). However, blastocyst quality, as evaluated by cryotolerance and gene expression analysis, was not found to be different between the groups. In conclusion, in vivo melatonin treatment is beneficial for increasing ovarian follicle turnover and improving oocyte developmental competence and kinetics of the blastocyst. PMID- 19552762 TI - Melatonin induces mitochondrial-mediated apoptosis in human myeloid HL-60 cells. AB - The role of melatonin in the mediation of apoptotic events has recently gained attention, especially after recent studies have reported that melatonin exerts antiapoptotic actions in normal cells but may activate proapoptotic pathways in some tumor cells. Here, we have evaluated the effect of melatonin on apoptosis in the human leukemia cell line HL-60. Melatonin treatment (1 mm) induced a significant increase in caspase-3 and -9 activities. The effect of melatonin on the activation of caspases was time dependent, reaching a maximum after 12 hr of stimulation, and then decreasing to a minimum after 72 hr. Treatment with melatonin also evoked mitochondrial membrane depolarization and permeability transition pore induction, which caused loss of mitochondrial staining by calcein, and increased cell death by apoptosis/necrosis as demonstrated by propidium iodide positive-staining of cells after 72 hr of stimulation. In addition, the exposure of cells to melatonin resulted in an activation and association of the proapoptotic proteins Bax and Bid, as well as promoting detectable increases in the expression of both proteins. We conclude that melatonin has proapoptotic and/or oncostatic effects in the human myeloid cell line HL-60. PMID- 19552763 TI - Melatonin ameliorates methionine- and choline-deficient diet-induced nonalcoholic steatohepatitis in rats. AB - Nonalcoholic steatohepatitis (NASH) may progress to advanced fibrosis and cirrhosis. Mainly, oxidative stress and excessive hepatocyte apoptosis are implicated in the pathogenesis of progressive NASH. Melatonin is not only a powerful antioxidant but also an anti-inflammatory and anti-apoptotic agent. We aimed to evaluate the effects of melatonin on methionine- and choline-deficient diet (MCDD)-induced NASH in rats. Thirty-two male Wistar rats were divided into four groups. Two groups were fed with MCDD while the other two groups were fed a control diet, pair-fed. One of the MCDD groups and one of the control diet groups were administered melatonin 50 mg/kg/day intraperitoneally, and the controls were given a vehicle. After 1 month the liver tissue oxidative stress markers, proinflammatory cytokines and hepatocyte apoptosis were studied by commercially available kits. For grading and staging histological lesions, Brunt et al.'s system was used. Melatonin decreased oxidative stress, proinflammatory cytokines and hepatocyte apoptosis. The drug ameliorated the grade of NASH. The present study suggests that melatonin functions as a potent antioxidant, anti inflammatory and antiapoptotic agent in NASH and may be a therapeutic option. PMID- 19552764 TI - Altered basal and postprandial plasma melatonin, gastrin, ghrelin, leptin and insulin in patients with liver cirrhosis and portal hypertension without and with oral administration of melatonin or tryptophan. AB - This investigation was designed to assess the effects of oral administration of melatonin (10 mg) and tryptophan (Trp) (500 mg) on fasting and postprandial plasma levels of melatonin, gastrin, ghrelin, leptin and insulin in 10 healthy controls and in age-matched patients with liver cirrhosis (LC) and portal hypertension. Fasting plasma melatonin levels in LC patients were about five times higher (102 +/- 15 pg/mL) than in healthy controls (22 +/- 3 pg/mL). These levels significantly increased postprandially in LC patients, but significantly less so in controls. Treatment with melatonin or L-Trp resulted in a further significant rise in plasma melatonin, both under fasting and postprandial conditions, particularly in LC patients. Moreover, plasma gastrin, ghrelin, leptin and insulin levels under fasting and postprandial conditions were significantly higher in LC subjects than in healthy controls and they further rose significantly after oral application of melatonin or Trp. This study shows that: (a) patients with LC and portal hypertension exhibit significantly higher fasting and postprandial plasma melatonin levels than healthy subjects; (b) plasma ghrelin, both in LC and healthy controls reach the highest values under fasting conditions, but decline postprandially, especially after oral application of melatonin or Trp; and (c) plasma melatonin, gastrin, ghrelin and insulin levels are altered significantly in LC patients with portal hypertension compared with that in healthy controls possibly due to their portal systemic shunting and decreased liver degradation. PMID- 19552765 TI - Melatonin down-regulates HIF-1 alpha expression through inhibition of protein translation in prostate cancer cells. AB - Melatonin, the main secretory product of the pineal gland, has been shown to exert an oncostatic activity in cancer cells. Recently, several studies have shown that melatonin has antiangiogenic properties. However, the mechanism by which melatonin exerts antiangiogenenic effects is not understood. Hypoxia inducible factor (HIF)-1 is a transcription factor which mediates adaptive response to changes in tissue oxygenation. HIF-1 is a heterodimer formed by the association of a constitutively expressed HIF-1 beta subunit and a HIF-1 alpha subunit, the expression of which is highly regulated. In this study, pharmacologic concentrations of melatonin was found to inhibit expression of HIF 1 alpha protein under both normoxic and hypoxic conditions in DU145, PC-3, and LNCaP prostate cancer cells without affecting HIF-1 alpha mRNA levels. Consistent with the reduction in HIF-1 alpha protein levels, melatonin inhibited HIF-1 transcriptional activity and the release of vascular endothelial growth factor. We found that the suppression of HIF-1 alpha expression by melatonin correlated with dephosphorylation of p70S6K and its direct target RPS6, a pathway known to regulate HIF-1 alpha expression at the translational level. Metabolic labeling assays indicated that melatonin inhibits de novo synthesis of HIF-1 alpha protein. Taken together, these results suggest that the pharmacologic concentration of melatonin inhibits HIF-1 alpha expression through the suppression of protein translation in prostate cancer cells. PMID- 19552767 TI - A comparative analysis of transarterial downstaging for hepatocellular carcinoma: chemoembolization versus radioembolization. AB - Chemoembolization and other ablative therapies are routinely utilized in downstaging from United Network for Organ Sharing (UNOS) T3 to T2, thus potentially making patients transplant candidates under the UNOS model for end stage liver disease (MELD) upgrade for hepatocellular carcinoma (HCC). This study was undertaken to compare the downstaging efficacy of transarterial chemoembolization (TACE) versus transarterial radioembolization. Eighty-six patients were treated with either TACE (n = 43) or transarterial radioembolization with Yttrium-90 microspheres (TARE-Y90; n = 43). Median tumor size was similar (TACE: 5.7 cm, TARE-Y90: 5.6 cm). Partial response rates favored TARE-Y90 versus TACE (61% vs. 37%). Downstaging to UNOS T2 was achieved in 31% of TACE and 58% of TARE-Y90 patients. Time to progression according to UNOS criteria was similar for both groups (18.2 months for TACE vs. 33.3 months for TARE-Y90, p = 0.098). Event-free survival was significantly greater for TARE-Y90 than TACE (17.7 vs. 7.1 months, p = 0.0017). Overall survival favored TARE-Y90 compared to TACE (censored 35.7/18.7 months; p = 0.18; uncensored 41.6/19.2 months; p = 0.008). In conclusion, TARE-Y90 appears to outperform TACE for downstaging HCC from UNOS T3 to T2. PMID- 19552766 TI - Coexpression of the melatonin receptor 1 and nestin in human breast cancer specimens. AB - Activation of the G-protein-coupled receptor (GPCR) for melatonin (MT1) suppresses breast cancer cell growth in experimental models. To elucidate whether MT1 might play a role in cancer cells positive for the stem cell marker nestin, we assessed paired carcinomatous (Ca) and adjacent noncancerous (NCa) samples from 42 patients with primary breast cancer for MT1 and nestin by double immunofluorescence staining and quantitative image analysis with Tissue-Quest software. MT1 was located in luminal and myoepithelial cells in milk ducts and in tumor cells in 40/42 and 39/42 of NCa and Ca specimens, respectively, independent of hormone receptor and HER-2 status. Nestin was located together with MT1 in myoepithelial cells in 38 NCa specimens (total n = 42) and in 18 Ca specimens with intact milk ducts. Quantitative evaluation of selected 16 NCa and Ca samples revealed that MT1 levels were higher in invasive Ca sections than in NCa specimens in eight and lower in six cases. Specimens from higher tumor stages (TII/III) with a higher risk of relapse were associated with MT1/nestin co staining in more than 10% of tumor cells, whereas a lack of co-staining correlated with lower tumor stages. Abundant expression of MT1 and, particularly, coexpression of MT1 with nestin in invading tumor cells in more advanced tumors suggest an important role for this GPCR in the pathogenesis of breast cancer. PMID- 19552768 TI - An imbalance in Akt/mTOR is involved in the apoptotic and acantholytic processes in a mouse model of pemphigus vulgaris. AB - Pemphigus vulgaris (PV) is an autoimmune blistering disease characterized by the presence of IgG autoantibodies against Dsg3. Our aim was to investigate the molecular events implicated in the development and localization of apoptosis and acantholysis in PV. We used a passive transfer mouse model together with immunohistochemical (IHC) techniques and the TUNEL assay, with quantification analysis in the basal layer of the epidermis. The activated signalling molecules analysed and apoptotic cells detected showed an identical localization. Herein, we found for the first time in vivo an increased expression of activated HER receptor isoforms in the basal layer in PV lesions. Besides, we observed the almost total lack of activated Akt compared with a higher level of activated mTOR within the basal cells of the epidermis. Our observations strongly support that the restriction of acantholysis to the basal layer may be due, at least in part, to the selective and increased presence of activated HER receptor isoforms in these cells. After phosphorylation of HER receptor isoforms, intracellular signalling pathways are activated in the basal layer. In addition, the imbalance in Akt/mTOR that takes place in the basal cells may provide intracellular signals necessary for the development of apoptosis and acantholysis. PMID- 19552769 TI - Establishment of a screening system for essential genes from the pathogenic yeast Candida glabrata: identification of a putative TEM1 homologue. AB - AIMS: To establish a system for screening and identification of essential genes from the pathogenic haploid yeast Candida glabrata by using temperature-sensitive (ts) mutants. METHODS AND RESULTS: Based on the general concepts that ts mutations are generated within essential genes in the genome by virtue of point mutation, we attempted to establish a system where essential genes were screened and identified from the C. glabrata genomic DNA library by the complementation of ts point mutations. By using this system, we successfully identified a putative TEM1 homologue as an essential gene by the complementation of a point mutation ( GAT-/-AAT- corresponding to Asp-143/Asn substitution) within its coding region in a ts mutant, T-3. CONCLUSIONS: We were able to establish a system for screening and identification of the essential genes, such as the TEM1 homologue, from the pathogenic yeast C. glabrata, as the gene that complements ts mutation. SIGNIFICANCE AND IMPACT OF THE STUDY: The identification of essential genes, by using the present system, may provide novel potential antifungal targets. PMID- 19552770 TI - Prevalence of thermotolerant Campylobacter in partridges (Perdix perdix). AB - AIM: To estimate the prevalence of thermotolerant Campylobacter spp. in commercially reared partridges (Perdix perdix) in southern Italy. METHODS AND RESULTS: Cloacal swabs of partridges (n = 240), equally distributed between male and female birds, from a game bird farm located in the Southern Italy were examined for the prevalence of thermotolerant Campylobacter spp. The samples were processed in order to detect thermotolerant Campylobacter spp. by culture methods. The positive samples were then confirmed by multiplex polymerase chain reaction. Thermotolerant Campylobacter spp. were isolated from 118 (49.2%) of the 240 cloacal swabs examined. As proved by PCR, 100% of the strains were identified as Campylobacter coli (118/118), and 15 (12.7%) out of the 118 positive samples were also positive for Campylobacter jejuni. In contrast, Campylobacter lari was not identified. Adult partridges showed a significantly higher prevalence (P < 0.05) than younger ones. CONCLUSION: These results reinforce the assumption that game birds may be considered as potential carriers of Campylobacter spp. for human being and other animal species. SIGNIFICANCE AND IMPACT OF THE STUDY: Although an earlier 1986 publication described the prevalence of Campylobacter coli in commercially reared partridges, this is the first report to confirm the species of Campylobacter using a PCR test. PMID- 19552771 TI - A DNA microarray facilitates the diagnosis of Bacillus anthracis in environmental samples. AB - AIMS: In order to improve the diagnosis of Bacillus anthracis in environmental samples, we established a DNA microarray based on the ArrayTube technology of Clondiag. METHODS AND RESULTS: Total DNA of a bacterial colony is randomly biotinylated and hybridized to the array. The probes on the array target the virulence genes, the genomic marker gene rpoB, as well as the selective 16S rDNA sequence regions of B. anthracis, of the Bacillus cereus group and of Bacillus subtilis. Eight B. anthracis reference strains were tested and correctly identified. Among the analysed environmental Bacillus isolates, no virulent B. anthracis strain was detected. CONCLUSIONS: This array clearly differentiates B. anthracis from members of the B. cereus group and other Bacillus species in environmental samples by chromosomal (rpoB) and plasmid markers. Additionally, recognition of B. cereus strains harbouring the toxin genes or atypical B. anthracis strains that have lost the virulence plasmids is feasible. SIGNIFICANCE AND IMPACT OF THE STUDY: The array is applicable to the complex diagnostics for B. anthracis detection in environmental samples. Because of low costs, high security and easy handling, the microarray is applicable to routine diagnostics. PMID- 19552772 TI - Genetic characterization of microbial communities living at the surface of building stones. AB - AIMS: The aim of the present study was to reveal the microbial genetic diversity of epilithic biofilms using a DNA-based procedure. METHODS AND RESULTS: A DNA extraction protocol was first selected to obtain PCR-amplifiable metagenomic DNA from a limestone biofilm. Extracted DNA was used to amplify either 16S rRNA genes or ITS regions from prokaryotic and eukaryotic genomes, respectively. Amplified DNAs were subsequently cloned, amplified by colony PCR and screened by restriction analysis [restriction analyses of amplified ribosomal DNA (ARDRA)] for DNA sequencing. Phylogenetic analysis using 16S rDNA sequences showed that predominating bacteria were Alphaproteobacteria belonging to the genera Sphingomonas, Erythrobacter, Porphyrobacter, Rhodopila and Jannashia; Cyanobacteria and Actinobacteria were also identified. Analysis of ITS rDNA sequences revealed the presence of algae of the Chlorophyceae family and fungi related either to Rhinocladiella or to a melanized ascomycete. Statistical analysis showed that the specific richness evidenced was representative of the original sampled biofilm. CONCLUSIONS: The molecular methodology developed here constitutes a valuable tool to investigate the genetic diversity of microbial biofilms from building stone. SIGNIFICANCE AND IMPACT OF THE STUDY: The easy-to run molecular method described here has practical importance to establish microbiological diagnosis and to define strategies for protection and restoration of stone surfaces. PMID- 19552774 TI - Prevalence of Campylobacter spp. in a subset of intensive poultry flocks in Ireland. AB - AIM: The aim of this study was to investigate the prevalence of Campylobacter species in a subset of intensive poultry flocks by examining samples collected in geographically disparate areas on the island of Ireland. METHODS AND RESULTS: Faecal, water and environmental samples were collected from the interior of poultry houses on nine farms. Three cultural methods were used for Campylobacter isolation: direct plating, enrichment culture and a recovery method for emerging Campylobacter spp. Presumptive Campylobacter isolates were confirmed using biochemical tests and further identified to species level by multiplex PCR. All flocks sampled in this study were found to be contaminated with Campylobacter at the time of sampling. Structural and air samples taken from the interior of broiler houses were also found to be Campylobacter positive. All water samples were found to be Campylobacter negative. The Campycheck method was used for the isolation of emerging Campylobacter spp. CONCLUSIONS: Campylobacter spp. were recovered (as contaminants) from the poultry house interior, air and environmental samples in all intensive poultry flocks surveyed. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the need for improved biosecurity on selected poultry farms. PMID- 19552773 TI - A combination of assays reveals biomass differences in biofilms formed by Escherichia coli mutants. AB - AIMS: The aim of this study was to develop an assay system that can quantify the amount of biomass in biofilms formed by different isogenic mutants of an Escherichia coli K-12 strain. METHODS AND RESULTS: The reported assay, which is based on the BacTiter-Glo assay from Promega, uses bioluminescence to detect the intracellular concentration of ATP, which correlates with viable bacterial cell numbers. The quantitative data obtained with this ATP assay were compared to those obtained with the conventional crystal violet assay. As a qualitative control, scanning electron microscopy was performed. CONCLUSIONS: The ATP assay, the crystal violet assay and scanning electron microscopy yielded similar results for six of the eight strains tested. For the remaining two strains, the images from the scanning electron microscopy confirmed the results from the ATP assay. SIGNIFICANCE AND IMPACT OF THE STUDY: The ATP assay, in combination with other quantitative and qualitative assays, will allow us to perform genetic studies on the regulatory network that underlies the early steps in E. coli biofilm formation. PMID- 19552775 TI - Isolation and characterization of actinomycetes from healthy goat faeces. AB - AIMS: To isolate and characterize actinomycetes with probiotic activities from healthy goat faeces. METHODS AND RESULTS: Faecal actinomycetes were isolated by dilution methods and identified by 16S rRNA gene sequence analysis. The hydrolytic enzyme activities were analysed by clear zone formation. The antimicrobial activities and resistance to heavy metals were tested by growth inhibition methods. The isolates belong to a small group of actinobacterial genera, including Streptomyces, Nocardiopsis and Oerskovia. The Oerskovia was the most widely distributed genus among the cultures. The proportion of streptomycete like strains producing amylase or protease is significantly higher than those of other actinomycetes (P < 0.05). Compared with streptomycete-like strains, a higher proportion of (alpha- or beta-) galactase-producing other actinomycetes was found in goat faeces. More than 50% of streptomycete-like strains showed activities against test fungi. Streptomycetes could tolerate 0.25 mmol l(-1) Cr(2)O(7)(2-), 2 mmol l(-1) Ni(2+); however, other actinomycetes are liable to 40 mmol l(-1) Fe(3+) and 0.25 mmol l(-1) Cr(2)O(7)(2-) and resistant to 5 mmol l(-1) Ni(2+) and 2 mmol l(-1) Cu(2+). CONCLUSIONS: The different physiological characteristics of the actinomycetes suggested that the cooperation in the actinomycetes might be involved in their association with goat. SIGNIFICANCE AND IMPACT OF THE STUDY: Probiotic mixtures based on faecal actinomycetes showed potentials in animal production. PMID- 19552776 TI - Characterization of probiotic carnobacteria isolated from rainbow trout (Oncorhynchus mykiss) intestine. AB - AIMS: To characterize two probiotic carnobacterial isolates, Carnobacterium maltaromaticum (B26) and C. divergens (B33), derived from rainbow trout (Oncorhynchus mykiss) intestine. METHODS AND RESULTS: Both cultures, which were able to colonize the fish gut mucosal layer, comprised nonsporogenous, nonmotile, Gram-positive, catalase and oxidase-negative rods. The growth of both carnobacteria occurred between 0 and 37 degrees C, in 0-10% (w/v) NaCl and at pH 5-10. Specifically, strain B26 grew in nutrient broth supplemented with 15% (w/v) NaCl. The most abundant cellular fatty acid of both cultures was 9-octadecenoic acid (18 : 1 n-9) (B26 = 52.6%; B33 = 40.6%), which was characteristic of Carnobacterium. Both cultures were inhibitory to Aeromonas salmonicida, Aer. hydrophila, Streptococcus iniae and Vibrio anguillarum, and strain B33 inhibited Listeria monocytogenes. Both carnobacteria, which did not contain plasmids, produced inhibitory compounds against Gram-positive and Gram-negative bacteria. CONCLUSIONS: Both probiotic cultures, B26 and B33, had unique phenotypic characteristics and showed a broad spectrum of antibiotic resistance against varying pathogenic bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study contribute to new information and significance of carnobacterial species. PMID- 19552777 TI - Cefixime-tellurite rhamnose MacConkey agar for isolation of Vero cytotoxin producing Escherichia coli serogroup O26 from Scottish cattle and sheep faeces. AB - AIMS: To compare rhamnose MacConkey agar supplemented with cefixime and tellurite (CT-RMac) and tryptone bile X-glucuronide (TBX) agars as isolation media for Vero cytotoxin-producing Escherichia coli (VTEC) serogroup O26 from animal faeces. METHODS AND RESULTS: Nine VTEC O26 were isolated from sheep faeces; out of which six were isolated only on CT-RMac and one was isolated only on TBX. One hundred and twelve VTEC O26 were isolated from calf faeces; out of which 97% were from CT RMac and 52% were from TBX. In a study of E. coli O26 strains, 84% of VT-positive O26 did not ferment rhamnose when compared with 16% of VT-negative O26. VT positive (19%) and VT-negative (39%) E. coli O26 strains did not grow on CT-RMac agar. CONCLUSIONS: It is important to consider that VTEC O26 strains either may ferment rhamnose or may be sensitive to the CT supplement of CT-RMac agar. SIGNIFICANCE AND IMPACT OF THE STUDY: This work compares CT-RMac and TBX agars as isolation medium for VTEC O26 from Scottish animal faeces and highlights that VTEC O26 may be missed if only CT-RMac agar is used. PMID- 19552778 TI - Genetic characterization of spoilage pseudomonads isolated from retail-displayed beef. AB - AIM: This study genetically characterized Pseudomonas isolated from beef using the random amplification of polymorphic DNA (RAPD) method and correlate predominant genotypes with spoilage changes. METHODS AND RESULTS: Pseudomonads were recovered from beef loins and steaks on days 0, 2, 4, 6, 8 and 10. A total of 309 pseudomonads were grouped into 50 RAPD types (>85% similarity). One major RAPD type contained 45% of the isolates comprising 71%, 45%, 31%, 35%, 50% and 37% of isolates from days 0, 2, 4, 6, 8 and 10, respectively, from steaks and 48% of the isolates recovered from beef loins. Nineteen RAPD types consisted of isolates that were shared between more than two sampling times, whereas the remaining 31 types were unique to one particular time. CONCLUSIONS: A genetically diverse Pseudomonas population was present on the loins and steaks at each sampling time. Although pseudomonads associated with beef loins were transferred to the steaks prepared from it, a genetically diverse Pseudomonas population emerged during the retail display. SIGNIFICANCE AND IMPACT OF THE STUDY: Information about the heterogeneous nature of Pseudomonas recovered from meat would help understanding the spoilage owing to predominant strains. The meat industry can use the knowledge to develop control strategies for prevalent spoilage strains. PMID- 19552779 TI - Evaluation of genome-derived amplicon length polymorphism PCR primers for the genetic evaluation of related strains of Salmonella. AB - AIMS: The intent of this study is to exploit both the genetic diversity and conservation demonstrated between the Salmonella enterica ssp. enterica serovar Typhi CT18 and Salm. enterica ssp. enterica serovar Typhimurium LT2 genomes by utilizing amplicon length polymorphism (ALP) to detect and differentiate various Salmonella strains. METHODS AND RESULTS: Methods of ALP-PCR analysis were developed based on identifying DNA sequence deletions within highly homologous regions of the Salm. Typhi CT18 and Salm. Typhimurium LT2 genomes. This study describes the application of genome-based ALP-PCR using primer pairs designed to detect genomic differences present within both Salmonella genomes and evaluated against a reference set of Salmonella strains. CONCLUSIONS: This study defines a collection of primer sequences broadly distributed along the Salmonella genome that can differentiate various Salmonella strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Genome-based ALP-PCR provides a useful and powerful analytical method to evaluate variability within a group of Salmonella strains independent of serological, phenotypic or other molecular approaches. PMID- 19552780 TI - Comparison of bacteriostatic and bactericidal activity of 13 essential oils against strains with varying sensitivity to antibiotics. AB - AIMS: To compare the bacteriostatic and bactericidal activity of 13 chemotyped essential oils (EO) on 65 bacteria with varying sensitivity to antibiotics. METHODS AND RESULTS: Fifty-five bacterial strains were tested with two methods used for evaluation of antimicrobial activity (CLSI recommendations): the agar dilution method and the time-killing curve method. EO containing aldehydes (Cinnamomum verum bark and Cymbopogon citratus), phenols (Origanum compactum, Trachyspermum ammi, Thymus satureioides, Eugenia caryophyllus and Cinnamomum verum leaf) showed the highest antimicrobial activity with minimum inhibitory concentration (MIC) <2% (v/v) against all strains except Pseudomonas aeruginosa. Alcohol-based EO (Melaleuca alternifolia, Cymbopogon martinii and Lavandula angustifolia) exhibited varying degrees of activity depending on Gram status. EO containing 1.8-cineole and hydrocarbons (Eucalyptus globulus, Melaleuca cajeputii and Citrus sinensis) had MIC(90%) > or = 10% (v/v). Against P. aeruginosa, only C. verum bark and O. compactum presented MIC < or =2% (v/v). Cinnamomum verum bark, O. compactum, T. satureioides, C. verum leaf and M. alternifolia were bactericidal against Staphylococcus aureus and Escherichia coli at concentrations ranging from to 0.31% to 10% (v/v) after 1 h of contact. Cinnamomum verum bark and O. compactum were bactericidal against P. aeruginosa within 5 min at concentrations <2% (v/v). CONCLUSIONS: Cinnamomum verum bark had the highest antimicrobial activity, particularly against resistant strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteriostatic and bactericidal activity of EO on nosocomial antibiotic-resistant strains. PMID- 19552781 TI - The antibacterial mechanism of carvacrol and thymol against Escherichia coli. AB - AIMS: To investigate the antibacterial mechanism of carvacrol and thymol against Escherichia coli. METHODS AND RESULTS: The time-kill curve results showed that carvacrol and thymol at 200 mg l(-1) could inhibit the growth of E. coli. Flow cytometry and fluorescent dyes were used to explore the effect of two components on membrane permeability and membrane potential. In membrane permeability experiment, the mean fluorescence intensity of cells treated with 200 mg l(-1) carvacrol or thymol were lower than nonexposed cells. The ratio of red to green fluorescence intensity of DiOC2(3) reflected the change of membrane potential. Carvacrol and thymol at 200 mg l(-1) caused the ratio of red/green decreasing from 0.42 of control to 0.08 and 0.07, respectively. CONCLUSIONS: Carvacrol and thymol had desired antimicrobial effect on E. coli. The antibacterial effects were attributed to their ability to permeabilize and depolarize the cytoplasmic membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: This study showed the potential use of flow cytometry as a suitable method to investigate the mode of antibacterial action of essential oil components. PMID- 19552782 TI - Isolation and characterization of lipopeptide antibiotics produced by Bacillus subtilis. AB - AIMS: Antibiotics from Bacillus subtilis JA show strong pathogen inhibition ability, which has potential market application; yet, the composition of these antibiotics has not been elucidated. The aim of this paper is to isolate and identify these antibiotics. METHODS AND RESULTS: The antagonistic activity of JA was tested in vitro; it exhibited strong inhibition against some important phytopathogens and postharvest pathogens. Crude antibiotic production was extracted with methanol from the precipitate by adding 6 mol l(-1) HCl to the bacillus-free culture broth. The crude extract was run on Diamonsil C18 column (5 microm, 250 x 4.6 mm) in HPLC system to separate the antibiotics. Major antibiotics were classified into three lipopeptide families according to electrospray ionization-mass spectrometry analysis. Subsequently, the classification of antibiotics was confirmed with typical collision-induced dissociation fragments. CONCLUSIONS: Three kinds of antibiotics were isolated from B. subtilis JA and were identified to the lipopeptide families, surfactin, iturin and fengycin. These compounds could function as biocontrol agents against a large spectrum of pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provided a reliable and rapid method for isolation and structural characterization of lipopeptide antibiotics from B. subtilis. PMID- 19552783 TI - Use of Hugh and Leifson's medium as a simple screening test to aid in the differentiation of Arcobacter spp. from background flora during their isolation from foodstuffs. AB - AIMS: To investigate the suitability of Hugh and Leifson's medium (HLM) as the basis of a simple screening test to differentiate between contaminants and Arcobacter spp. during their isolation from foodstuffs. METHODS AND RESULTS: Characterized Arcobacter spp. were obtained from recognized culture collections. Wild-type isolates of Arcobacter spp. and contaminants were obtained using published isolation protocols. Retail packs of red meats were used as the source of the isolates. Eighteen defined Arcobacter spp. gave no reaction on HLM, as did 10 local wild-type isolates. Overall 163 contaminants were studied for oxidative reactions on HLM and 86% of isolates demonstrated this property. CONCLUSIONS: HLM can usefully serve as a simple and effective screening test to differentiate between Arcobacter spp. and contaminants. SIGNIFICANCE AND IMPACT OF THE STUDY: Arcobacter isolation procedures are still being developed, and no effective diagnostic media currently exist. Rapidly excluding most contaminants can markedly increase the efficiency of isolation procedures by removing the need for extensive biotyping or the requirement to isolate DNA and conduct PCR tests. PMID- 19552784 TI - Effect of using a stand-off pad on Campylobacter jejuni strain diversity in a herd of dairy cows. AB - AIM: To determine the effect of stand-off pad (SOP) use on the prevalence and strain diversity of Campylobacter jejuni in a small herd of dairy cows. METHODS AND RESULTS: Faecal samples were collected from 21 cows on four sampling occasions (events), one in each season, over 1 year. The cows usually grazed on pasture but during winter they spent 18 h a day on a SOP. Campylobacter prevalence ranged from 48-52% on pasture but was 62% on the SOP. The diversity of 386 C. jejuni isolates was determined using Enterobacterial Repetitive Intergenic Consensus polymerase chain reaction (ERIC/PCR). There were 11 ERIC types identified for the herd over the course of the study. Of those 11, four to seven (per event) were present when the cows were grazing pasture but only two during SOP use. CONCLUSIONS: The use of the SOP was associated with an increase in prevalence and a reduction in diversity of C. jejuni. SIGNIFICANCE AND IMPACT OF THE STUDY: The reduction in ERIC types on the SOP indicated an increase in transfer of only some strains of C. jejuni among the cows. One of these strains persisted throughout the study. The zoonotic potential of this strain warrants further investigation. PMID- 19552785 TI - OTA-producing fungi isolated from stored cocoa beans. AB - AIMS: The aim of this study was to identify fungal populations in unroasted cocoa beans stored in Spain in order to evaluate the ochratoxin A (OTA)-production ability of certain Aspergillus isolates. METHODS AND RESULTS: Twenty batches of cocoa beans from different origins and with different OTA content were selected for this study. Three Aspergillus carbonarius and 13 Aspergillus niger aggregate strains isolated from these cocoa bean samples were selected to evaluate their OTA synthesis ability, being the only A. carbonarius isolates which are OTA producers [or= 2 mg/L increased from 19.4% to 35.5%. In conclusion, we describe the alternation of MRSA clones that occurred in hospitals from 1990 to 2007 and the increase of the glycopeptide MIC levels, reflecting a worldwide trend. We document the detection of ST1, ST8, ST15 and ST30 in the 1980 isolates; we hypothesize their possible latency and their appearance as the current CA-MRSA clones. PMID- 19552803 TI - Characterization and isolation of a T-DNA tagged banana promoter active during in vitro culture and low temperature stress. AB - BACKGROUND: Next-generation transgenic plants will require a more precise regulation of transgene expression, preferably under the control of native promoters. A genome-wide T-DNA tagging strategy was therefore performed for the identification and characterization of novel banana promoters. Embryogenic cell suspensions of a plantain-type banana were transformed with a promoterless, codon optimized luciferase (luc+) gene and low temperature-responsive luciferase activation was monitored in real time. RESULTS: Around 16,000 transgenic cell colonies were screened for baseline luciferase activity at room temperature 2 months after transformation. After discarding positive colonies, cultures were re screened in real-time at 26 degrees C followed by a gradual decrease to 8 degrees C. The baseline activation frequency was 0.98%, while the frequency of low temperature-responsive luciferase activity was 0.61% in the same population of cell cultures. Transgenic colonies with luciferase activity responsive to low temperature were regenerated to plantlets and luciferase expression patterns monitored during different regeneration stages. Twenty four banana DNA sequences flanking the right T-DNA borders in seven independent lines were cloned via PCR walking. RT-PCR analysis in one line containing five inserts allowed the identification of the sequence that had activated luciferase expression under low temperature stress in a developmentally regulated manner. This activating sequence was fused to the uidA reporter gene and back-transformed into a commercial dessert banana cultivar, in which its original expression pattern was confirmed. CONCLUSION: This promoter tagging and real-time screening platform proved valuable for the identification of novel promoters and genes in banana and for monitoring expression patterns throughout in vitro development and low temperature treatment. Combination of PCR walking techniques was efficient for the isolation of candidate promoters even in a multicopy T-DNA line. Qualitative and quantitative GUS expression analyses of one tagged promoter in a commercial cultivar demonstrated a reproducible promoter activity pattern during in vitro culture. Thus, this promoter could be used during in vitro selection and generation of commercial transgenic plants. PMID- 19552804 TI - Transcriptional profiling in response to terminal drought stress reveals differential responses along the wheat genome. AB - BACKGROUND: Water stress during grain filling has a marked effect on grain yield, leading to a reduced endosperm cell number and thus sink capacity to accumulate dry matter. The bread wheat cultivar Chinese Spring (CS), a Chinese Spring terminal deletion line (CS_5AL-10) and the durum wheat cultivar Creso were subjected to transcriptional profiling after exposure to mild and severe drought stress at the grain filling stage to find evidences of differential stress responses associated to different wheat genome regions. RESULTS: The transcriptome analysis of Creso, CS and its deletion line revealed 8,552 non redundant probe sets with different expression levels, mainly due to the comparisons between the two species. The drought treatments modified the expression of 3,056 probe sets. Besides a set of genes showing a similar drought response in Creso and CS, cluster analysis revealed several drought response features that can be associated to the different genomic structure of Creso, CS and CS_5AL-10. Some drought-related genes were expressed at lower level (or not expressed) in Creso (which lacks the D genome) or in the CS_5AL-10 deletion line compared to CS. The chromosome location of a set of these genes was confirmed by PCR-based mapping on the D genome (or the 5AL-10 region). Many clusters were characterized by different level of expression in Creso, CS and CS_AL-10, suggesting that the different genome organization of the three genotypes may affect plant adaptation to stress. Clusters with similar expression trend were grouped and functional classified to mine the biological mean of their activation or repression. Genes involved in ABA, proline, glycine-betaine and sorbitol pathways were found up-regulated by drought stress. Furthermore, the enhanced expression of a set of transposons and retrotransposons was detected in CS_5AL 10. CONCLUSION: Bread and durum wheat genotypes were characterized by a different physiological reaction to water stress and by a substantially different molecular response. The genome organization accounted for differences in the expression level of hundreds of genes located on the D genome or controlled by regulators located on the D genome. When a genomic stress (deletion of a chromosomal region) was combined with low water availability, a molecular response based on the activation of transposons and retrotransposons was observed. PMID- 19552805 TI - Optimising corticosteroid injection for lateral epicondylalgia with the addition of physiotherapy: a protocol for a randomised control trial with placebo comparison. AB - BACKGROUND: Corticosteroid injection and physiotherapy are two commonly prescribed interventions for management of lateral epicondylalgia. Corticosteroid injections are the most clinically efficacious in the short term but are associated with high recurrence rates and delayed recovery, while physiotherapy is similar to injections at 6 weeks but with significantly lower recurrence rates. Whilst practitioners frequently recommend combining physiotherapy and injection to overcome harmful effects and improve outcomes, study of the benefits of this combination of treatments is lacking. Clinicians are also faced with the paradox that the powerful anti-inflammatory corticosteroid injections work well, albeit in the short term, for a non-inflammatory condition like lateral epicondylalgia. Surprisingly, these injections have not been rigorously tested against placebo injections. This study primarily addresses both of these issues. METHODS: A randomised placebo-controlled clinical trial with a 2 x 2 factorial design will evaluate the clinical efficacy, cost-effectiveness and recurrence rates of adding physiotherapy to an injection. In addition, the clinical efficacy and adverse effects of corticosteroid injection beyond that of a placebo saline injection will be studied. 132 participants with a diagnosis of lateral epicondylalgia will be randomly assigned by concealed allocation to one of four treatment groups - corticosteroid injection, saline injection, corticosteroid injection with physiotherapy or saline injection with physiotherapy. Physiotherapy will comprise 8 sessions of elbow manipulation and exercise over an 8 week period. Blinded follow-up assessments will be conducted at baseline, 4, 8, 12, 26 and 52 weeks after randomisation. The primary outcome will be a participant rating of global improvement, from which measures of success and recurrence will be derived. Analyses will be conducted on an intention-to-treat basis using linear mixed and logistic regression models. Healthcare costs will be collected from a societal perspective, and along with willingness-to-pay and quality of life data will facilitate cost-effectiveness and cost-benefit analyses. CONCLUSION: This trial will utilise high quality trial methodologies in accordance with CONSORT guidelines. Findings from this study will assist in the development of evidence based practice recommendations and potentially the optimisation of resource allocation for rehabilitating lateral epicondylalgia. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Register ACTRN12609000051246. PMID- 19552806 TI - Production and characterisation of monoclonal antibodies against RAI3 and its expression in human breast cancer. AB - BACKGROUND: RAI3 is an orphan G-protein coupled receptor (GPCR) that has been associated with malignancy and may play a role in the proliferation of breast cancer cells. Although its exact function in normal and malignant cells remains unclear and evidence supporting its role in oncogenesis is controversial, its abundant expression on the surface of cancer cells would make it an interesting target for the development of antibody-based therapeutics. To investigate the link with cancer and provide more evidence for its role, we carried out a systematic analysis of RAI3 expression in a large set of human breast cancer specimens. METHODS: We expressed recombinant human RAI3 in bacteria and reconstituted the purified protein in liposomes to raise monoclonal antibodies using classical hybridoma techniques. The specific binding activity of the antibodies was confirmed by enzyme-linked immunosorbent assay (ELISA), western blot and immunocytochemistry. We carried out a systematic immunohistochemical analysis of RAI3 expression in human invasive breast carcinomas (n = 147) and normal breast tissues (n = 44) using a tissue microarray. In addition, a cDNA dot blot hybridisation assay was used to investigate a set of matched normal and cancerous breast tissue specimens (n = 50) as well as lymph node metastases (n = 3) for RAI3 mRNA expression. RESULTS: The anti-RAI3 monoclonal antibodies bound to recombinant human RAI3 protein with high specificity and affinity, as shown by ELISA, western blot and ICC. The cDNA dot blot and immunohistochemical experiments showed that both RAI3 mRNA and RAI3 protein were abundantly expressed in human breast carcinoma. However, there was no association between RAI3 protein expression and prognosis based on overall and recurrence-free survival. CONCLUSION: We have generated a novel, highly-specific monoclonal antibody that detects RAI3 in formaldehyde-fixed paraffin-embedded tissue. This is the first study to report a systematic analysis of RAI3 expression in normal and cancerous human breast tissue at both the mRNA and protein levels. PMID- 19552807 TI - Reliability and validity of a new scale on internal coherence (ICS) of cancer patients. AB - BACKGROUND: Current inventories on quality of life used in oncology mainly focus on functional aspects of patients in the context of disease adaptation and treatments (side) effects (EORTC QLQ C30) or generically the status of common functions (Medical Outcome Study SF 36). Beyond circumscribed dimensions of quality of life (i.e., physical, emotional, social, cognitive etc.), there is a lack of inventories which also address other relevant dimensions such as the 'sense of coherence' (SOC) in cancer patients. SOC is important because of its potential prognostic relevance in cancer patients, but the current SOC scale has mainly been validated for psychiatric and psychosomatic patients. Our two-step validation study addresses the internal coherence (ICS) scale, which is based on expert rating, using specific items for oncological patients, with respect to its reliability, validity and sensitivity to chemotherapy. METHODS: The items were tested on 114 participants (57 cancer patients and a matched control group), alongside questions on autonomic regulation (aR), the Hospital Anxiety and Depression Scale (HADS), self-regulation (SRQ) and Karnofsky the Performance Index (KPI). A retest of 65 participants was carried out after a median time span of four weeks.In the second part of the study, the ICS was used to assess internal coherence during chemotherapy in 25 patients with colorectal carcinoma (CRC) and 17 breast cancer patients. ICS was recorded before, during and 4-8 weeks after treatment. RESULTS: The 10-item scale of 'internal coherence' (ICS) shows good to very good reliability: Cronbach-alpha r = 0.91, retest-reliability r = 0.80. The ICS correlates with r = 0.43-0.72 to the convergence criteria (all p < 0.001). We are able to show decreased ICS-values after the third cycle for CRC and breast cancer patients, with a subsequent increase of ICS scores after the end of chemotherapy. CONCLUSION: The ICS has good to very good reliability, validity and sensitivity to chemotherapy. PMID- 19552810 TI - The effect of a brief social intervention on the examination results of UK medical students: a cluster randomised controlled trial. AB - BACKGROUND: Ethnic minority (EM) medical students and doctors underperform academically, but little evidence exists on how to ameliorate the problem. Psychologists Cohen et al. recently demonstrated that a written self-affirmation intervention substantially improved EM adolescents' school grades several months later. Cohen et al.'s methods were replicated in the different setting of UK undergraduate medical education. METHODS: All 348 Year 3 white (W) and EM students at one UK medical school were randomly allocated to an intervention condition (writing about one's own values) or a control condition (writing about another's values), via their tutor group. Students and assessors were blind to the existence of the study. Group comparisons on post-intervention written and OSCE (clinical) assessment scores adjusted for baseline written assessment scores were made using two-way analysis of covariance. All assessment scores were transformed to z-scores (mean = 0 standard deviation = 1) for ease of comparison. Comparisons between types of words used in essays were calculated using t-tests. The study was covered by University Ethics Committee guidelines. RESULTS: Groups were statistically identical at baseline on demographic and psychological factors, and analysis was by intention to treat [intervention group EM n = 95, W n = 79; control group EM n = 77; W n = 84]. As predicted, there was a significant ethnicity by intervention interaction [F(4,334) = 5.74; p = 0.017] on the written assessment. Unexpectedly, this was due to decreased scores in the W intervention group [mean difference = 0.283; (95% CI = 0.093 to 0.474] not improved EM intervention group scores [mean difference = -0.060 (95% CI = -0.268 to 0.148)]. On the OSCE, both W and EM intervention groups outperformed controls [mean difference = 0.261; (95%CI = -0.047 to -0.476; p = 0.013)]. The intervention group used more optimistic words (p < 0.001) and more "I" and "self" pronouns in their essays (p < 0.001), whereas the control group used more "other" pronouns (p < 0.001) and more negations (p < 0.001). DISCUSSION: Cohen et al.'s finding that a brief self-affirmation task narrowed the ethnic academic achievement gap was replicated on the written assessment but against expectations, this was due to reduced performance in the W group. On the OSCE, the intervention improved performance in both W and EM groups. In the intervention condition, participants tended to write about themselves and used more optimistic words than in the control group, indicating the task was completed as requested. The study shows that minimal interventions can have substantial educational outcomes several months later, which has implications for the multitude of seemingly trivial changes in teaching that are made on an everyday basis, whose consequences are never formally assessed. PMID- 19552809 TI - Highly focused anopheline breeding sites and malaria transmission in Dakar. AB - BACKGROUND: Urbanization has a great impact on the composition of the vector system and malaria transmission dynamics. In Dakar, some malaria cases are autochthonous but parasite rates and incidences of clinical malaria attacks have been recorded at low levels. Ecological heterogeneity of malaria transmission was investigated in Dakar, in order to characterize the Anopheles breeding sites in the city and to study the dynamics of larval density and adult aggressiveness in ten characteristically different urban areas. METHODS: Ten study areas were sampled in Dakar and Pikine. Mosquitoes were collected by human landing collection during four nights in each area (120 person-nights). The Plasmodium falciparum circumsporozoite (CSP) index was measured by ELISA and the entomological inoculation rates (EIR) were calculated. Open water collections in the study areas were monitored weekly for physico-chemical characterization and the presence of anopheline larvae. Adult mosquitoes and hatched larvae were identified morphologically and by molecular methods. RESULTS: In September October 2007, 19,451 adult mosquitoes were caught among which, 1,101 were Anopheles gambiae s.l. The Human Biting Rate ranged from 0.1 bites per person per night in Yoff Village to 43.7 in Almadies. Seven out of 1,101 An. gambiae s.l. were found to be positive for P. falciparum (CSP index = 0.64%). EIR ranged from 0 infected bites per person per year in Yoff Village to 16.8 in Almadies. The An. gambiae complex population was composed of Anopheles arabiensis (94.8%) and Anopheles melas (5.2%). None of the An. melas were infected with P. falciparum. Of the 54 water collection sites monitored, 33 (61.1%) served as anopheline breeding sites on at least one observation. No An. melas was identified among the larval samples. Some physico-chemical characteristics of water bodies were associated with the presence/absence of anopheline larvae and with larval density. A very close parallel between larval and adult densities was found in six of the ten study areas. CONCLUSION: The results provide evidence of malaria transmission in downtown Dakar and its surrounding suburbs. Spatial heterogeneity of human biting rates was very marked and malaria transmission was highly focal. In Dakar, mean figures for transmission would not provide a comprehensive picture of the entomological situation; risk evaluation should therefore be undertaken on a small scale. PMID- 19552811 TI - The potential monetary benefits of reclaiming hazardous waste sites in the Campania region: an economic evaluation. AB - BACKGROUND: Evaluating the economic benefit of reducing negative health outcomes resulting from waste management is of pivotal importance for designing an effective waste policy that takes into account the health consequences for the populations exposed to environmental hazards. Despite the high level of Italian and international media interest in the problem of hazardous waste in Campania little has been done to reclaim the land and the waterways contaminated by hazardous waste. OBJECTIVE: This study aims to reduce the uncertainty about health damage due to waste exposure by providing for the first time a monetary valuation of health benefits arising from the reclamation of hazardous waste dumps in Campania. METHODS: First the criteria by which the landfills in the Campania region, in particular in the two provinces of Naples and Caserta, have been classified are described. Then, the annual cases of premature death and fatal cases of cancers attributable to waste exposure are quantified. Finally, the present value of the health benefits from the reclamation of polluted land is estimated for each of the health outcomes (premature mortality, fatal cancer and premature mortality adjusted for the cancer premium). Due to the uncertainty about the time frame of the benefits arising from reclamation, the latency of the effects of toxic waste on human health and the lack of context specific estimates of the Value of Preventing a Fatality (VPF), extensive sensitivity analyses are performed. RESULTS: There are estimated to be 848 cases of premature mortality and 403 cases of fatal cancer per year as a consequence of exposure to toxic waste. The present value of the benefit of reducing the number of waste associated deaths after adjusting for a cancer premium is euro11.6 billion. This value ranges from euro5.4 to euro20.0 billion assuming a time frame for benefits of 10 and 50 years respectively. CONCLUSION: This study suggests that there is a strong economic argument for both reclaiming the land contaminated with hazardous waste in the two provinces of Naples and Caserta and increasing the control of the territory in order to avoid the creation of new illegal dump sites. PMID- 19552812 TI - Impact of animal strain on gene expression in a rat model of acute cardiac rejection. AB - BACKGROUND: The expression levels of many genes show wide natural variation among strains or populations. This study investigated the potential for animal strain related genotypic differences to confound gene expression profiles in acute cellular rejection (ACR). Using a rat heart transplant model and 2 different rat strains (Dark Agouti, and Brown Norway), microarrays were performed on native hearts, transplanted hearts, and peripheral blood mononuclear cells (PBMC). RESULTS: In heart tissue, strain alone affected the expression of only 33 probesets while rejection affected the expression of 1368 probesets (FDR 10% and FC > o= 3). Only 13 genes were affected by both strain and rejection, which was < 1% (13/1368) of all probesets differentially expressed in ACR. However, for PBMC, strain alone affected 265 probesets (FDR 10% and FC > or = 3) and the addition of ACR had little further effect. Pathway analysis of these differentially expressed strain effect genes connected them with immune response, cell motility and cell death, functional themes that overlap with those related to ACR. After accounting for animal strain, additional analysis identified 30 PBMC candidate genes potentially associated with ACR. CONCLUSION: In ACR, genetic background has a large impact on the transcriptome of immune cells, but not heart tissue. Gene expression studies of ACR should avoid study designs that require cross strain comparisons between leukocytes. PMID- 19552813 TI - Data publication: towards a database of everything. AB - The fabric of science is changing, driven by a revolution in digital technologies that facilitate the acquisition and communication of massive amounts of data. This is changing the nature of collaboration and expanding opportunities to participate in science. If digital technologies are the engine of this revolution, digital data are its fuel. But for many scientific disciplines, this fuel is in short supply. The publication of primary data is not a universal or mandatory part of science, and despite policies and proclamations to the contrary, calls to make data publicly available have largely gone unheeded. In this short essay I consider why, and explore some of the challenges that lie ahead, as we work toward a database of everything. PMID- 19552808 TI - Construction of an adult barnacle (Balanus amphitrite) cDNA library and selection of reference genes for quantitative RT-PCR studies. AB - BACKGROUND: Balanus amphitrite is a barnacle commonly used in biofouling research. Although many aspects of its biology have been elucidated, the lack of genetic information is impeding a molecular understanding of its life cycle. As part of a wider multidisciplinary approach to reveal the biogenic cues influencing barnacle settlement and metamorphosis, we have sequenced and annotated the first cDNA library for B. amphitrite. We also present a systematic validation of potential reference genes for normalization of quantitative real time PCR (qRT-PCR) data obtained from different developmental stages of this animal. RESULTS: We generated a cDNA library containing expressed sequence tags (ESTs) from adult B. amphitrite. A total of 609 unique sequences (comprising 79 assembled clusters and 530 singlets) were derived from 905 reliable unidirectionally sequenced ESTs. Bioinformatics tools such as BLAST, HMMer and InterPro were employed to allow functional annotation of the ESTs. Based on these analyses, we selected 11 genes to study their ability to normalize qRT-PCR data. Total RNA extracted from 7 developmental stages was reverse transcribed and the expression stability of the selected genes was compared using geNorm, BestKeeper and NormFinder. These software programs produced highly comparable results, with the most stable gene being mt-cyb, while tuba, tubb and cp1 were clearly unsuitable for data normalization. CONCLUSION: The collection of B. amphitrite ESTs and their annotation has been made publically available representing an important resource for both basic and applied research on this species. We developed a qRT-PCR assay to determine the most reliable reference genes. Transcripts encoding cytochrome b and NADH dehydrogenase subunit 1 were expressed most stably, although other genes also performed well and could prove useful to normalize gene expression studies. PMID- 19552814 TI - Single-copy nuclear genes resolve the phylogeny of the holometabolous insects. AB - BACKGROUND: Evolutionary relationships among the 11 extant orders of insects that undergo complete metamorphosis, called Holometabola, remain either unresolved or contentious, but are extremely important as a context for accurate comparative biology of insect model organisms. The most phylogenetically enigmatic holometabolan insects are Strepsiptera or twisted wing parasites, whose evolutionary relationship to any other insect order is unconfirmed. They have been controversially proposed as the closest relatives of the flies, based on rDNA, and a possible homeotic transformation in the common ancestor of both groups that would make the reduced forewings of Strepsiptera homologous to the reduced hindwings of Diptera. Here we present evidence from nucleotide sequences of six single-copy nuclear protein coding genes used to reconstruct phylogenetic relationships and estimate evolutionary divergence times for all holometabolan orders. RESULTS: Our results strongly support Hymenoptera as the earliest branching holometabolan lineage, the monophyly of the extant orders, including the fleas, and traditionally recognized groupings of Neuropteroidea and Mecopterida. Most significantly, we find strong support for a close relationship between Coleoptera (beetles) and Strepsiptera, a previously proposed, but analytically controversial relationship. Exploratory analyses reveal that this relationship cannot be explained by long-branch attraction or other systematic biases. Bayesian divergence times analysis, with reference to specific fossil constraints, places the origin of Holometabola in the Carboniferous (355 Ma), a date significantly older than previous paleontological and morphological phylogenetic reconstructions. The origin and diversification of most extant insect orders began in the Triassic, but flourished in the Jurassic, with multiple adaptive radiations producing the astounding diversity of insect species for which these groups are so well known. CONCLUSION: These findings provide the most complete evolutionary framework for future comparative studies on holometabolous model organisms and contribute strong evidence for the resolution of the 'Strepsiptera problem', a long-standing and hotly debated issue in insect phylogenetics. PMID- 19552817 TI - Needs assessment of Wisconsin primary care residents and faculty regarding interest in global health training. AB - BACKGROUND: The primary objectives of this study were to assess Wisconsin's primary care residents' attitudes toward international health training, the interest among faculty to provide IH training, and the preferred modality of IH training. METHODS: Surveys were administered using 505 residents and 413 medical faculty in primary care residencies in Wisconsin. Results from 128 residents and 118 medical school faculty members were collected during the spring of 2007 and analyzed. RESULTS: In total, 25% of residents (128/505) and 28% of faculty (118/413) responded to the survey. A majority of residents (58%) and faculty (63%) were interested in global health issues. Among residents, 63% planned on spending professional time working abroad. Few residents (9%) and faculty (11%) assess their residencies as preparing residents well to address topics relating to international health. The survey indicates that adequate faculty in Wisconsin could provide mentorship in international health as 47% (55) of faculty had experience working as a physician internationally, 49% (58) of faculty spend more than 25% clinical time caring for patient from underserved communities and 39% (46) would be willing to be involved with developing curriculum, lecturing and/or mentoring residents in international health. CONCLUSION: Overall, the majority of the respondents expressed high interest in IH and few felt prepared to address IH issues indicating a need for increased training in this area. The findings of this survey are likely relevant as a prototype for other primary care residencies. PMID- 19552816 TI - Expression of multidrug resistance markers ABCB1 (MDR-1/P-gp) and ABCC1 (MRP-1) in renal cell carcinoma. AB - BACKGROUND: Renal cell carcinoma patients respond poorly to conventional chemotherapy, this unresponsiveness may be attributable to multidrug resistance (MDR). The mechanisms of MDR in renal cancer are not fully understood and the specific contribution of ABC transporter proteins which have been implicated in the chemoresistance of various cancers has not been fully defined in this disease. METHODS: In this retrospective study the expression of two of these transporter efflux pumps, namely MDR-1 P-gp (ABCB1) and MRP-1 (ABCC1) were studied by immunohistochemistry in archival material from 95 renal cell carcinoma patients. RESULTS: In the first study investigating MDR-1 P-gp and MRP-1 protein expression patterns in renal cell carcinoma patients, high levels of expression of both efflux pumps are observed with 100% of tumours studied showing MDR-1 P-gp and MRP-1 positivity. CONCLUSION: Although these findings do not prove a causal role, the high frequency of tumours expressing these efflux pumps suggests that they may be important contributors to the chemoresistance of this tumour type. PMID- 19552815 TI - Chronic crude garlic-feeding modified adult male rat testicular markers: mechanisms of action. AB - BACKGROUND: Garlic or Allium sativum (As) shows therapeutic effects such as reduction of blood pressure or hypercholesterolemia but side-effects on reproductive functions remain poorly investigated. Because of garlic's chemical complexity, the processing methods and yield in preparations differ in efficacy and safety. In this context, we clarify the mechanisms of action of crushed crude garlic on testicular markers. METHODS: During one month of treatment, 24 male rats were fed 5%, 10% and 15% crude garlic. RESULTS: We showed that crude garlic feeding induced apoptosis in testicular germ cells (spermatocytes and spermatids). This cell death process was characterized by increased levels of active CASP3 but not CASP6. Expression of the caspase inhibitors BIRC3 and BIRC2 was increased at all doses of As while expression of XIAP and BIRC5 was unchanged. Moreover, expression of the IAP inhibitor DIABLO was increased at doses 10% and 15% of As. The germ cell death process induced by As might be related to a decrease in testosterone production because of the reduced expression of steroidogenic enzymes (Star, Cyp11a, Hsd3b5 and Hsd17b). Evaluation of Sertoli markers showed that TUBB3 and GSTA2 expression was unchanged. In contrast, AMH, RHOX5 and CDKN1B expression was decreased while GATA4 expression was increased. CONCLUSION: In summary, we showed that feeding with crude garlic inhibited Leydig steroidogenic enzyme expression and Sertoli cell markers. These alterations might induce apoptosis in testicular germ cells. PMID- 19552818 TI - Activin A induces ovine follicle stimulating hormone beta using -169/-58 bp of its promoter and a simple TATA box. AB - BACKGROUND: Activin A increases production of follicle stimulating hormone (FSH) by inducing transcription of its beta subunit (FSHB). This induction has been studied here in LbetaT2 gonadotropes using transient expression of ovine FSHBLuc (-4741 bp of ovine FSHB promoter plus exon/intron 1 linked to Luc). Several sequences between -169/-58 bp of the ovine FSHB proximal promoter are necessary for induction by activin A in LbetaT2 cells, but deletions between -4741/-752 bp decrease induction > 70% suggesting the existence of other important 5' sequences. Induction disappears if a minimal T81 thymidine kinase promoter replaces the ovine FSHB TATA box and 3' exon/intron. The study reported here was designed to determine if sequences outside -169/-58 bp are important for induction of ovine FSHB by activin A. METHODS: Progressively longer deletions of ovine FSHBLuc were created between -4741/-195 bp. Deletions internal to this region were created also, but replaced with substitute DNA. The ovine FSHB TATA box region (-40/+3 bp) was replaced by thymidine kinase and rat prolactin minimal promoters, and substitutions were made in 3' intron/exon sequences. All constructs were tested for basal and activin A-induced expression in LbetaT2 cells. RESULTS: Successive 5' deletions progressively lowered fold-induction by activin A from 9.5 to zero, but progressively increased basal expression. Replacing deletions with substitute DNA showed no changes in basal expression or fold-induction. Induction by activin A was supported by the minimal rat prolactin promoter (TATA box) but not the thymidine kinase promoter (no TATA box). Replacement mutations in the 3' region did not decrease induction by activin A. CONCLUSION: The data show that specific ovine FSHB sequences 5' to -175 bp or 3' of the transcription start site are not required for induction by activin A. A minimal TATA box promoter supports induction by activin A, but the sequence between the TATA box and transcription start site seems unimportant. PMID- 19552819 TI - Trends in broad-spectrum antibiotic prescribing for children with acute otitis media in the United States, 1998-2004. AB - BACKGROUND: Overuse of broad-spectrum antibiotics is associated with antibiotic resistance. Acute otitis media (AOM) is responsible for a large proportion of antibiotics prescribed for US children. Rates of broad-spectrum antibiotic prescribing for AOM are unknown. METHODS: Analysis of the National Ambulatory Medical Care Survey and National Hospital Ambulatory Medical Care Survey, 1998 to 2004 (N = 6,878). Setting is office-based physicians, hospital outpatient departments, and emergency departments. Patients are children aged 12 years and younger prescribed antibiotics for acute otitis media. Main outcome measure is percentage of broad-spectrum antibiotics, defined as amoxicillin/clavulanate, macrolides, cephalosporins and quinolones. RESULTS: Broad-spectrum prescribing for acute otitis media increased from 34% of visits in 1998 to 45% of visits in 2004 (P < .001 for trend). The trend was primarily attributable to an increase in prescribing of amoxicillin/clavulanate (8% to 15%; P < .001 for trend) and macrolides (9% to 15%; P < .001 for trend). Prescribing remained stable for amoxicillin and cephalosporins while decreasing for narrow-spectrum agents (12% to 3%; P < .001 for trend) over the study period. Independent predictors of broad spectrum antibiotic prescribing were ear pain, non-white race, public and other insurance (compared to private), hospital outpatient department setting, emergency department setting, and West region (compared to South and Midwest regions), each of which was associated with lower rates of broad-spectrum prescribing. Age and fever were not associated with prescribing choice. CONCLUSION: Prescribing of broad-spectrum antibiotics for acute otitis media has steadily increased from 1998 to 2004. Associations with non-clinical factors suggest potential for improvement in prescribing practice. PMID- 19552821 TI - Ultrasonic partial glossectomy. AB - INTRODUCTION: Partial glossectomy is the main treatment for tongue carcinoma. The resection of the tongue, which is a very vascularised tissue, requires a good hemostasis. The advantage of the harmonic scalpel is in combining sectioning and hemostasis in one single instrument, allowing a bloodless dissection of soft tissue. The aim of this prospective study was to evaluate the benefits and risks when using a harmonic scalpel in partial glossectomy. SUBJECTS AND METHODS: In this prospective study conducted in a university hospital from march 2004 to December 2008, eighteen consecutive patients underwent a partial glossectomy with the use of harmonic scalpel. Results were compared with previous surgical procedures performed between September 2000 and February 2004 by monopolar hemostasis by our team (n = 12) when the harmonic scalpel was not available. RESULTS: All 18 patients underwent partial glossectomy with the harmonic scalpel as the only instrument of section and hemostasis. The median blood loss was of 0 mL. The median operative time was 29 minutes (16 minutes less than partial glossectomies performed with conventional hemostasis. P < .001). No operative complications occurred. Two post-operative bleedings (5 days and 7 days after the glossectomy) occurred necessitating a new surgery to ligate the lingual artery. The margins of the resection were acceptable and no recurrence appeared. CONCLUSION: The harmonic scalpel makes it fast and easy to perform a partial glossectomy with no bleeding. Ligation of the lingual artery (when it is visualized during the dissection) should be performed because of the frequency (more than 10% in our series) and because of the potential gravity of a lingual post-operative bleeding. PMID- 19552820 TI - Epidermal wound healing in severe sepsis and septic shock in humans. AB - INTRODUCTION: The effect of sepsis on epidermal wound healing has not been previously studied. It was hypothesised that epidermal wound healing is disturbed in severe sepsis. METHODS: Blister wounds were induced in 35 patients with severe sepsis and in 15 healthy controls. The healing of the wounds was followed up by measuring transepidermal water loss and blood flow in the wound, reflecting the restoration of the epidermal barrier function and inflammation, respectively. The first set of suction blisters (early wound) was made within 48 hours of the first sepsis-induced organ failure and the second set (late wound) four days after the first wound. In addition, measurements were made on the intact skin. RESULTS: The average age of the whole study population was 62 years (standard deviation [SD] 12). The mean Acute Physiology and Chronic Health Evaluation II (APACHE II) score on admission was 25 (SD 8). The two most common causes of infections were peritonitis and pneumonia. Sixty-six percent of the patients developed multiple organ failure. The decrease in water evaporation from the wound during the first four days was lower in septic patients than in the control subjects (56 g/m2 per hour versus 124 g/m2 per hour, P = 0.004). On the fourth day, septic patients had significantly higher blood flow in the wound compared with the control subjects (septic patients 110 units versus control subjects 47 units, P = 0.001). No difference in transepidermal water loss from the intact skin was found between septic patients and controls. Septic patients had higher blood flow in the intact skin on the fourth and on the eighth day of study compared with the controls. CONCLUSIONS: The restoration of the epidermal barrier function is delayed and wound blood flow is increased in patients with severe sepsis. PMID- 19552822 TI - Exploring spatial patterns and hotspots of diarrhea in Chiang Mai, Thailand. AB - BACKGROUND: Diarrhea is a major public health problem in Thailand. The Ministry of Public Health, Thailand, has been trying to monitor and control this disease for many years. The methodology and the results from this study could be useful for public health officers to develop a system to monitor and prevent diarrhea outbreaks. METHODS: The objective of this study was to analyse the epidemic outbreak patterns of diarrhea in Chiang Mai province, Northern Thailand, in terms of their geographical distributions and hotspot identification. The data of patients with diarrhea at village level and the 2001-2006 population censuses were collected to achieve the objective. Spatial analysis, using geographic information systems (GIS) and other methods, was used to uncover the hidden phenomena from the data. In the data analysis section, spatial statistics such as quadrant analysis (QA), nearest neighbour analysis (NNA), and spatial autocorrelation analysis (SAA), were used to identify the spatial patterns of diarrhea in Chiang Mai province. In addition, local indicators of spatial association (LISA) and kernel density (KD) estimation were used to detect diarrhea hotspots using data at village level. RESULTS: The hotspot maps produced by the LISA and KD techniques showed spatial trend patterns of diarrhea diffusion. Villages in the middle and northern regions revealed higher incidences. Also, the spatial patterns of diarrhea during the years 2001 and 2006 were found to represent spatially clustered patterns, both at global and local scales. CONCLUSION: Spatial analysis methods in GIS revealed the spatial patterns and hotspots of diarrhea in Chiang Mai province from the year 2001 to 2006. To implement specific and geographically appropriate public health risk-reduction programs, the use of such spatial analysis tools may become an integral component in the epidemiologic description, analysis, and risk assessment of diarrhea. PMID- 19552823 TI - Physicians' attitudes about obesity and their associations with competency and specialty: a cross-sectional study. AB - BACKGROUND: Physicians frequently report negative attitudes about obesity which is thought to affect patient care. However, little is known about how attitudes toward treating obese patients are formed. We conducted a cross-sectional survey of physicians in order to better characterize their attitudes and explore the relationships among attitudes, perceived competency in obesity care, including report of weight loss in patients, and other key physician, training, and practice characteristics. METHODS: We surveyed all 399 physicians from internal medicine, pediatrics, and psychiatry specialties at one institution regarding obesity care attitudes, competency, including physician report of percent of their patients who lose weight. We performed a factor analysis on the attitude items and used hierarchical regression analysis to explore the degree to which competency, reported weight loss, physician, training and practice characteristics explained the variance in each attitude factor. RESULTS: The overall response rate was 63%. More than 40% of physicians had a negative reaction towards obese patients, 56% felt qualified to treat obesity, and 46% felt successful in this realm. The factor analysis revealed 4 factors-Physician Discomfort/Bias, Physician Success/Self Efficacy, Positive Outcome Expectancy, and Negative Outcome Expectancy. Competency and reported percent of patients who lose weight were most strongly associated with the Physician Success/Self Efficacy attitude factor. Greater skill in patient assessment was associated with less Physician Discomfort/Bias. Training characteristics were associated with outcome expectancies with newer physicians reporting more positive treatment expectancies. Pediatric faculty was more positive and psychiatry faculty less negative in their treatment expectancies than internal medicine faculty. CONCLUSION: Physician attitudes towards obesity are associated with competency, specialty, and years since postgraduate training. Further study is necessary to determine the direction of influence and to explore the impact of these attitudes on patient care. PMID- 19552824 TI - Klassevirus 1, a previously undescribed member of the family Picornaviridae, is globally widespread. AB - BACKGROUND: Diarrhea is the third leading infectious cause of death worldwide and is estimated to be responsible for approximately 2 million deaths a year. While many infectious causes of diarrhea have been established, approximately 40% of all diarrhea cases are of unknown etiology. In an effort to identify novel viruses that may be causal agents of diarrhea, we used high throughput mass sequencing to analyze stool samples collected from patients with acute diarrhea. RESULTS: Sequences with limited similarity to known picornaviruses were detected in a stool sample collected in Australia from a child with acute diarrhea. Using a combination of mass sequencing, RT-PCR, 5' RACE and 3' RACE, a 6383 bp fragment of the viral genome was sequenced. Phylogenetic analysis demonstrated that this virus was highly divergent from, but most closely related to, members of the genus Kobuvirus. We have tentatively named this novel virus klassevirus 1. We also detected klassevirus 1 by RT-PCR in a diarrhea specimen collected from a patient in St. Louis, United States as well as in untreated sewage collected in Barcelona, Spain. CONCLUSION: Klassevirus 1 is a previously undescribed picornavirus that is globally widespread and present on at least three continents. Further investigations to determine whether klassevirus 1 is a human pathogen are needed. PMID- 19552825 TI - Characterization of Zur-dependent genes and direct Zur targets in Yersinia pestis. AB - BACKGROUND: The zinc uptake regulator Zur is a Zn2+-sensing metalloregulatory protein involved in the maintenance of bacterial zinc homeostasis. Up to now, regulation of zinc homeostasis by Zur is poorly understood in Y. pestis. RESULTS: We constructed a zur null mutant of Y. pestis biovar microtus strain 201. Microarray expression analysis disclosed a set of 154 Zur-dependent genes of Y. pestis upon exposure to zinc rich condition. Real-time reverse transcription (RT) PCR was subsequently used to validate the microarray data. Based on the 154 Zur dependent genes, predicted regulatory Zur motifs were used to screen for potential direct Zur targets including three putative operons znuA, znuCB and ykgM-RpmJ2. The LacZ reporter fusion analysis verified that Zur greatly repressed the promoter activity of the above three operons. The subsequent electrophoretic mobility shift assay (EMSA) demonstrated that a purified Zur protein was able to bind to the promoter regions of the above three operons. The DNase I footprinting was used to identify the Zur binding sites for the above three operons, verifying the Zur box sequence as predicted previously in gamma-Proteobacteria. The primer extension assay was further used to determine the transcription start sites for the above three operons and to localize the -10 and -35 elements. Zur binding sites overlapped the -10 sequence of its target promoters, which was consistent with the previous observation that Zur binding would block the entry of the RNA polymerase to repress the transcription of its target genes. CONCLUSION: Zur as a repressor directly controls the transcription of znuA, znuCB and ykgM-RpmJ2 in Y. pestis by employing a conserved mechanism of Zur-promoter DNA association as observed in gamma-Proteobacteria. Zur contributes to zinc homeostasis in Y. pestis likely through transcriptional repression of the high-affinity zinc uptake system ZnuACB and two alternative ribosomal proteins YkgM and RpmJ2. PMID- 19552826 TI - Multidirectional instability of the shoulder - current concept. AB - A guest editorial on the multidirectional instability of the shoulder. PMID- 19552827 TI - HIV serostatus disclosure pattern among pregnant women in Enugu, Nigeria. AB - This study was carried out in two medical facilities in Enugu, Nigeria, from September to November 2007. An interviewer-administered questionnaire was used to collect data from HIV-positive pregnant women accessing PMTCT (prevention of maternal-to-child transmission) services at the two centres. Ninety-two women were interviewed: 89 (96.7%) had disclosed their status, while 3 (3.3%) had not. Of the 89 women who had disclosed, 84 (94.4%) had disclosed to partners, 82 (92.1%) to husbands, 2 (2.2%) to fiances, 18 (20.2%) to sisters, 13 (14.6%) to mothers, 10 (11.2%) to brothers, 10 (11.2%) to fathers and 10 (11.2%) to priests. Fifty-two (58.4%) gave emotional support as the reason for disclosure and 46 (51.7%) gave economic and financial support as reasons. Fifty-six (62.9%) reported understanding from partner as a positive outcome and 44 (49.4%) reported financial support. Forty-six (51.7%) reported no negative outcome. Serostatus disclosure rate in this study was high with most women disclosing to their partners. PMID- 19552828 TI - Infant and child mortality among Catholics and Lutherans in nineteenth century Poznan. AB - The purpose of this study was to show the differences in the mortality rates of children from Catholic and Lutheran families in 19th century Poznan, and to elucidate the causes of these differences. Data from Catholic and Lutheran parish death registers were used. The infant death rate (IDR), neonatal and postneonatal death rates and life table biometric functions were calculated and causes of deaths were characterized. The worst child mortality values (IDR=394.4; neonatal and postneonatal death rates, respectively, 117.1 and 277.4; e0=16.14 years; Crow's Index=2.47) were obtained for the poor Catholic Parish of St Margaret. The lowest infant and neonatal and postneonatal death rates were observed to have occurred in the Catholic Parish of St Maria Magdalena situated in the city's more affluent central area (mortality rates, respectively, 269.9, 93.1 and 176.9; e0=24.63 years; Crow's Index=0.96). The widest range of differences with regard to death rates was found for the Lutheran Parish of St Cross (the infant, neonatal and postneonatal death rates were, respectively, 293.1, 99.1 and 193.9; e0=28.03 years; Crow's Index=0.92). The St Cross Parish encompassed a fairly large area of the city characterized by varying ecological conditions. Among infants and young children from the three studied populations a high frequency of deaths due to infectious diseases, diarrhoeas, dysenteries and tuberculosis were observed. Differences in the mortality of children from Catholic and Lutheran families in 19th century Poznan resulted from ecological conditions, among which water played the most important role, rather than from religious differences. PMID- 19552829 TI - Fruit and vegetable intake and risk of upper respiratory tract infection in pregnant women. AB - OBJECTIVE: The present study evaluated the association between fruit and vegetable intake and the incidence of upper respiratory tract infection (URTI) during pregnancy. DESIGN: In a cohort of 1034 North American women, each subject was asked retrospectively about their fruit and vegetable intake during the six months before the pregnancy and their occurrences of URTI during the first half of pregnancy. Multivariable-adjusted hazard ratios (HR) were calculated with Cox proportional hazards models. RESULTS: The adjusted HR of URTI for women in the highest quartile (median 8.54 servings/d) v. the lowest quartile (median 1.91 servings/d) of total fruit and vegetable intake was 0.74 (95 % CI 0.53, 1.05) for the 5-month follow-up period and 0.61 (95 % CI 0.39, 0.97) for the 3-month follow up period, respectively. A dose-related reduction of URTI risk according to quartile of intake was found in the 3-month (P for trend = 0.03) but not the 5 month follow-up. No association was found between either fruit or vegetable intake alone in relation to the 5-month or the 3-month risk of URTI. CONCLUSIONS: Women who consume more fruits and vegetables have a moderate reduction in risk of URTI during pregnancy, and this benefit appears to be derived from both fruits and vegetables instead of either alone. PMID- 19552830 TI - The relationship between elderly suicide rates, population density and room density. PMID- 19552831 TI - Efficacy of SSRIs on cognition of Alzheimer's disease patients treated with cholinesterase inhibitors. AB - BACKGROUND: This study examines the joint effect on cognition of selective serotonin re-uptake inhibitors (SSRIs) and cholinesterase inhibitors (AChEIs) in depressed patients affected by Alzheimer's disease (AD) living at home. METHODS: The study was conducted in two different outpatient neurological clinics. 338 patients with probable AD were treated with ChEis (donepezil, rivastigmine and galantamine) as per the clinician's judgment and were observed for nine months. At study entry, participants underwent a multidimensional assessment evaluating cognitive, functional and psychobehavioral domains. All patients were evaluated at baseline, after one (T1), three (T2) and nine months (T3). Patients were grouped in three different categories (patients not depressed and not treated with SSRIs, patients depressed and treated with SSRIs, and patients depressed but not treated with SSRIs). RESULTS: At baseline 182 were diagnosed as not depressed and not treated with SSRIs, 66 as depressed and treated with SSRIs, and 90 as depressed but not treated with SSRIs. The mean change in MMSE score from baseline to nine months showed that depressed patients not treated worsened in comparison with those not depressed and not treated with SSRIs (mean change -0.8 +/- 2.3 vs 0.04 +/- 2.9; p = 0.02) and patients depressed and treated with SSRI (mean change -0.8 +/- 2.3 vs 0.1 +/- 2.5; p = 0.03). CONCLUSIONS: In AD patients treated with AChEIs, SSRIs may exert some degree of protection against the negative effects of depression on cognition. PMID- 19552832 TI - The early experience of Old Age Psychiatrists in the application of the Mental Capacity Act 2005: a pilot study. AB - BACKGROUND: The Mental Capacity Act 2005 (MCA) was fully implemented in October 2007 in England and Wales. METHODS: A pilot questionnaire study examined the experience of consultants in Old Age Psychiatry in the early implementation of the MCA pertaining to local policy and training in the application of the MCA, the assessment of decision-making capacity, the determination of best interests, and the use of the least restrictive option and restraint. RESULTS: Fifty-two (27%) of the 196 consultants in Old Age Psychiatry returned useable questionnaires. Seventy-five percent of them reported that local training on the application of the MCA was available, but less than 50% reported that training was mandatory. The vast majority of assessments of decision-making capacity were conducted by consultants in Old Age Psychiatry. Almost all of them reported using the four-fold specific test of decision-making capacity (DMC) described in the MCA. Restraint was reported to be rarely used. CONCLUSIONS: Consultants in Old Age Psychiatry generally reported using the criteria for the assessment of DMC, the determination of best interests and restraint described in the MCA. The findings highlight concern about the workload of clinicians in implementing the MCA and this requires careful monitoring. Consideration should be given to statutory provision of training in the application of the MCA by all healthcare and social care providers for all their healthcare and social care staff. PMID- 19552833 TI - The application of the Mental Capacity Act 2005 among geriatric psychiatry patients: a pilot study. AB - BACKGROUND: The Mental Capacity Act 2005 (MCA) was fully implemented in October 2007 in England and Wales. The experience of clinicians working in Old Age Psychiatry, who are likely to have greater experience in the use of MCA, in the early implementation of the MCA was examined. METHODS: Case-notes of 37 patients in seven different Old Age Psychiatry services in two mental health trusts in west London, who had received at least one assessment of decision-making capacity (DMC) for a specific issue, were examined. A qualitative thematic analysis pertaining to the criteria used for the assessment of DMC, determination of best interests, least restrictive option and unwise decision was used for data analysis. RESULTS: The main findings were: the criteria used for the assessment of DMC and the determination of best interests were those described in the MCA and the accompanying Code of Practice; and clinicians were developing the concepts of least restrictive option and unwise decision with face validity despite the absence of their definitions in the MCA. CONCLUSION: Caution should be exercised in extrapolating the findings of this study, which is confined to two Mental Health Trusts in one geographical area and the speciality of Old Age Psychiatry, to other localities and other specialties. Nevertheless, there was evidence that clinicians were following the basic principles of the MCA correctly. PMID- 19552834 TI - [Progress of society and science and technology promotes the development of gastrointestinal surgery]. PMID- 19552835 TI - [Major molecular response to imatinib in a patient with acute mixed lineage leukemia expressing a novel BCR/ABL transcript]. AB - OBJECTIVE: To identify the novel BCR/ABL transcript in a patient with acute mixed lineage leukemia (AMLL), and to evaluate the imatinib treatment response by quantitatively monitoring the aberrant BCR/ABL. METHODS: Specimens of peripheral blood and bone marrow were obtained. By using several detect protocols, we found a novel BCR/ABL transcript in a patient with Ph-positive AMLL. The patient was treated with Imatinib and the aberrant BCR/ABL was quantitatively monitored to evaluate the clinical response. RESULTS: On admission, cytogenetic analysis showed Philadelphia-chromosome (Ph) positive in the specimens, but BCR/ABL e1a2, b2a2, and b3a2 were negative. Morphological analysis of the bone marrow showed the myeloid blast cells accounted for 66%, and immunophenotyping analysis showed 2 groups of aberrant blast cells: myeloid and B lineage. Chemical therapy and bone marrow transplantation failed to control the disease, and a novel BCR/ABL transcript (GenBank: EF423615) was found by using several detection protocols. The novel fusion protein showed a deletion of 10 amino acids and H893Q compared with the common BCR/ABL b2a2 fusion protein. The patient was then treated with imatinib and hematological remission was soon achieved, and 5 months after the imatinib treatment the quantity of the aberrant BCR/ABL was gradually decreased to negative. During the treatment the patient had discontinued the drug once and just then the aberrant BCR/ABL became positive soon. Imatinib was administered again and molecular remission was soon achieved for the second time. By continued therapy with imatinib, the patient got sustained and complete molecular remission lasting 12 months so far. CONCLUSION: The aberrant BCR/ABL may contribute to the clinical features of AMLL and the AMLL patients that have aberrant BCR/ABL may be sensitive to Imatinib. PMID- 19552836 TI - [Methyl-CpG-binding protein 2 gene and CDKL5 gene mutation in patients with Rett syndrome: analysis of 177 Chinese pediatric patients]. AB - OBJECTIVE: To study the spectrum of mutations in methyl-CpG-binding protein 2 gene (MECP2) and cyclin-dependent kinase-like 5 gene (CDKL5) in Chinese pediatric patients with Rett syndrome (RTT), and establish a simple, quick, and efficient gene test method as well as screen a strategy of genetic diagnosis for RTT. METHODS: Genomic DNA was extracted using standard procedures from the peripheral blood leukocytes of 117 pediatric patients diagnosed from 1987 to 2007. PCR was used to amplify the exons 1 - 4 of MECP2 using published primers. If no mutation was identified after screening exons 2 - 4, exon 1 was screened. If no mutation was identified in MECP2 by sequencing, multiplex ligation dependent probe amplification (MLPA) was employed to screen for large deletions by using P015C kit. If no mutation was identified in the MECP2 by sequencing and MLPA respectively, then the coding region of CDKL5 was screened by denaturing high performance liquid chromatography (DHPLC). RESULTS: The total mutation frequency in MECP2 and CDKL5 genes among all RTT patients was 82%. MECP2 mutations were found in 86% (137/159) of the patients with classical RTT and in 44% (8/18) of those with atypical RTT. Most of the mutations were missense mutations, accounting for 39%, followed in order of frequency by nonsense mutations 28%, frame shift mutations 17% and large deletions 14.5%. The eight most frequent MECP2 mutations were p.T158M (13%), p.R168X (12%), c.806delG (7%), p.R255X (6%), p.R270X (5%), p.R133C (5%), p.R306C (4%), and p.R106W (3%), with p.T158M as the most common of the MECP2 mutations and c.806delG as a hotspot mutation in Chinese patients with RTT. Only one synonymous mutation was identified in CDKL5. CONCLUSION: The spectrum of MECP2 mutations within the mainland Chinese RTT patients is similar to that of those patients reported in the world. p.T158M, p.R168X, c.806delG, p.R255X, p.R270X, p.R133C, p.R306C, and p.R106W are the hotspot mutations of MECP2 and c.806delG is a specific hotspot mutation in Chinese patients with RTT. The most effective method to screen mutations is to screen the exon 4. MLPA is an effective supplement to the routine methods. PMID- 19552837 TI - [Bone morphogenetic protein type II receptor gene promoter mutation-142G > A in a patient with familial pulmonary arterial hypertension]. AB - OBJECTIVE: To investigate the relation between bone morphogenetic protein type II receptor (BMPR2) gene promoter mutation and pulmonary arterial hypertension (PAH). METHODS: Peripheral blood samples were collected from a 36-year-old female patient with familial PAH (FPAH), 19 idiopathic PAH (IPAH) patients, and 50 healthy controls. DNA sequencing was conducted for the position -2022 bp upstream of the promoter transcription start point of BMPR2 gene. Two fragments carrying BMPR2 promoter mutation -142A and wild -142G allele were amplified and cloned respectively into the pGL3-basic dual-luciferase reporter gene vector, thus generating two luciferase reporter constructs: pGL3-BMPR2-wild recombinant plasmid (carrying -142G allele) and pGL3-BMPR2-mut recombinant plasmid (carrying 142A allele). Human pulmonary arterial smooth muscle cells (HPASMCs) and human pulmonary arterial endothelial cells (HPAECs) were cultured and transfected with pGL3-BMPR2-wild and pGL3-BMPR2-mut recombinant plasmids respectively. The transcriptional activity levels of these 2 recombinant plasmids were measured by Veritas Microplate Luminometer, and were calculated as the ratio of firefly luciferase activity to Renilla luciferase activity. The binding sites for transcriptional factors on the flanking sequence of the wild and mutant BMPR2 gene promoter regions were analyzed by using the MAPPER Search Engine. RESULTS: A mutation -142G > A in the promoter region of BMPR2 gene was found in this female patient with FPAH. The transcriptional activity levels of the BMPR2 promoter carrying -142A allele in the HPASMCs and HPAECs were (9.58 +/- 3.85) and (59.07 +/- 25.54) respectively, both significantly lower than those of the BMPR2 promoter carrying -142G allele [(16.80 +/- 3.55) and (115.58 +/- 38.02) respectively, both P < 0.05]. The binding site of specificity protein 3, the potential transcriptional factor, was deleted in the BMPR2 promoter carrying 142A allele compared to the BMPR2 promoter carrying -142G allele. CONCLUSION: BMPR2 promoter mutation -142G > A may be associated with FPAH. PMID- 19552838 TI - [Relationship of serum Chemerin to obesity and type 2 diabetes mellitus]. AB - OBJECTIVE: To detect the serum Chemerin levels in patients with obesity and newly diagnosed type 2 diabetes mellitus (T2DM) and explore the relationship of serum Chemerin to body fat parameters, glucose and lipid metabolism, and insulin resistance index (IR). METHODS: 76 newly diagnosed T2DM patients and 76 subjects with normal glucose regulation (NGR), with the body mass index (BMI) < 25 kg/m(2) (NW) or > or = 25 kg/m(2) (OW/OB), 38 in each subgroups each, underwent body measurement including body weight, height, waist circumference, and hip circumference, and waist hip ratio (WHR) and calculation of body mass index (BMI) were calculated. Peripheral blood samples were collected from them to detect the blood lipids, glucose, hemoglobin A1C, fasting insulin (FINS), and fasting C peptide. Homeostasis model assessment insulin resistance index (HOMA-IR) was calculated. ELISA was used to detect the Chemerin level, body mass index (BMI) and waist hip ratio (WHR) were evaluated and insulin sensitivity was assessed by HOMA-IR. RESULTS: The serum Chemerin level of the females was (109 +/- 28) microg/L, significantly higher than that of the males [(98 +/- 23) microg/L, P < 0.05]. After adjustment of gender and age, the serum Chemerin level of the OW/OB group, including NGR-OW/OB and T2DM-OW/OB subgroups, was (113 +/- 27) microg/L, significantly higher than that of the NW group [(94 +/- 25) microg/L, P < 0.01], that of the NGR-OW/OB subgroup being the highest. Partial correlation analyses showed that the serum Chemerin was positively correlated with waist circumference, WHR, fasting serum C peptide, HOMA-IR, TG, ALT, gamma-GT, and uric acid (r = 0.460 - 0.182, all P < 0.05) and negatively correlated with high density lipoprotein-cholesterol (r = -0.251, P < 0.01). Stepwise regression analysis showed that fasting serum C peptide and TG were the independent variables of Chemerin (beta = 0.328, 0.280, P < 0.05). CONCLUSION: Serum Chemerin levels are much higher in females and obese subjects are much higher than in men and subjects with normal weight. Serum Chemerin is correlated with insulin level, body fat disposition and lipid metabolism which suggesting that it may play a role in the pathophysiology of obesity and metabolic syndrome. PMID- 19552839 TI - [Candida colonization and invasive fungal infection in hospitalized patients with hematological malignancies]. AB - OBJECTIVE: To investigate thee status of Candida colonization and the risk factors of invasive fungal infection (IFI) in hospitalized patients with hematological malignancies. METHODS: 114 patients with hematological malignancies admitted to the hematology wards from May 2004 to April 2005 underwent fungal culture of their samples of urine, feces, and saliva once a week until the end of the sixth week of hospitalization or they were discharged. Culture of blood, sputum, or sterile body fluids were carried out when the patients were suspected to have IFI. RESULTS: 165 strains of Candida spp. were isolated from 46 of the 114 patients, C. albicans accounting for 78.8% and non-albicans Candida for 21.2% respectively. Candida was found in 38 patients (33.3%) were found to have colonization of Candida, chiefly C. albicans (76.3%). The risk factors of Candida colonization included long duration of leucopenia (WBC < 0.5 x 10(9)/L for more than 10 days) and broad-spectrum antibiotic administration. Univariate analysis showed that the risk factors of IFI included Candida colonization, long duration of leucopenia, and administration of carbapenem antibiotics, and multivariate analysis showed that long duration of leucopenia (WBC < 0.5 x 10(9)/L for more than 10 days) was the only risk factor of IFI (P = 0.015). CONCLUSION: C. albicans is still the major organism isolated from the patients with hematological malignancies. Patients with hematological malignancies have a high incidence of Candida colonization. Long duration of leucopenia is the only risk factor for IFI. PMID- 19552840 TI - [Clinical-pathological characteristics and survival of triple negative breast cancer]. AB - OBJECTIVE: To investigate the clinical-pathological characteristics of triple negative breast cancer. METHODS: The clinical-pathological characteristics of 690 breast cancer patients, all females, 127 being estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor (HER)-2 triple negative, were analyzed. RESULTS: The expression rate of P53, a tumor suppressor gene, of the triple negative breast cancer patients was 71.42% (75/127), significantly higher than that of the non-triple negative breast cancer patients (42.56%, P < 0.01), and the expression rate of epidermal growth factor receptor (EGFR) of the triple negative breast cancer was 59.74%, significantly higher than that of the non-triple negative breast cancer patients (22.06%, P = 0.013). The local lymph node metastasis rate of the triple negative breast cancer was 22.22%, significantly higher than that of the non-triple negative breast cancer patients (2.70%, P = 0.048). The 5-year overall survival (OS) rate and 10 year OS rate of he triple negative breast cancer patients were 79.67% and 63.15% respectively, both significantly lower than those of the non-triple negative breast cancer patients (88.59% and 83.28% respectively, both P = 0.001). The 5 year and 10-year disease free survival (DFS) rates of the triple negative breast cancer patients were 77.94% and 62.87% respectively, both significantly lower than those of the non-triple negative breast cancer patients (83.82% and 82.53% respectively, both P = 0.011), especially that within 36 months. CONCLUSION: Triple negative breast cancer is related with P53 and EGFR expression, and tends to metastasize to local lympho-nodes. The survival rate of triple negative breast cancer patients is significantly lower than that of non-triple negative breast cancer. PMID- 19552841 TI - [Clinical character of renal transplant recipients with bilateral native upper urinary transitional cell carcinoma]. AB - OBJECTIVE: To analysis the clinical character of renal transplant (RTx) recipients with bilateral native upper urinary transitional cell carcinoma. METHODS: The clinical data of 7 cases of transitional cell carcinoma in 2236 RTx recipients who underwent non-synchronous native bilateral nephroureterectomy (BNU) for 14 case-times were retrospectively analyzed. RESULTS: The mean time between RTx and diagnosis of TCC was 49.7 months (24 - 96 months). The mean interval between 2 upper urinary tract operations was 14.0 months (2 - 36 months). All specimens obtained in the 14 operations were TCC-positive. The TCC location involved pure native pelvis (n = 2), pure native ureter (n = 4), and pelvis combined with ureter (n = 8). The TCC pathological grades include G(1) G(2) (n = 7), G3 (n = 6), and cancer in situ (n = 1). The TCC stage status included pTIS (n = 1), pT(1) N(0)M(0) (n = 4), pT(2)N(0)M(0) (n = 6), pT(3)N(0)M(0) (n = 2), and pT(4)N(2)M(1) (n = 1). Five patients were observed to have TCC in bladder. Each patient underwent more then 2 times of transurethral resection of bladder tumor (TUR-Bt). The bladder TCC was at G(2)-G(3) with deep of superficial muscle involvement. Metastasis was found in skin, soft tissue, and costal bone of lumbar in one patient. The blood creatinine level of the 7 patients, representing the graft's function, was 120 micromol/L (97 - 161 micromol/L). The patients were followed up for 2 - 48 months, all patients survived. CONCLUSION: TCC after RTx has multiple sources and is invasive. Non synchronous prophylactic native BNU with removal of bladder cuffs should be considered for those RTx recipients having multiple urinary tract malignancy in native upper urinary tract and bladder. PMID- 19552842 TI - [Correlation of mycophenolic acid pharmacokinetic parameters with clinical events in kidney transplant patients treated with mycophenolate mofetil]. AB - OBJECTIVE: To investigate the correlation of pharmacokinetic profiles of mycophenolic acid (MPA) with clinic events in kidney transplant patients treated with mycophenolate mofetil (MMF). METHODS: Sixty-four kidney transplant patients were treated with the triple immune suppression protocol with cyclosporine, MMF, and prednisone. Before the administration of drugs, and 0.5, 1, 1.5, 2, 2.5, 4, 6, 8, 10, and 12 hours after the drug administration peripheral blood samples were collected to detect the plasma MPA concentrations (MPA-C(0)) and draw the concentration-time curve. And 4, 7, 21, and 28 days, and 1.5, 2, 3, and 6 months later peripheral blood samples were collected in the morning before drug administration so as to dynamically measure the plasma MPA concentrations. Clinical events were observed. RESULTS: No side effect or acute rejection episode occurred with the median MPA-C(0) value of 0.74 mg/L. Side effects occurred with the median MPA-C(0) value of 1.32 mg/L. Sixteen case-times demonstrated acute rejection episode with the median MPA-C(0) value of 0.39 mg/L. The incidence rates of different side effects, such as infection, bone marrow depression, and digestive tract symptoms were significantly related to MPA-C(0) dose-dependently. CONCLUSION: MPA-C(0) may be an appropriate pharmacokinetic monitoring parameter for kidney transplant. PMID- 19552843 TI - [Meta-analysis of antibiotic prophylaxis for postoperative bacteriuria in patients undergoing transrectal prostatic biopsy]. AB - OBJECTIVE: To investigate whether antibiotic prophylaxis can reduce the risk of postoperative bacteriuria in patients undergoing transrectal prostatic biopsy (TPB) who have sterile preoperative urine. METHODS: MEDLINE, EMBASE, Cochrane Collaboration Reviews, CMCC, and CNKI were searched for randomized controlled trials (RCTs) comparing antibiotic prophylaxis with placebo/blank controls for patients undergoing TPB with preoperative sterile urine. The search strategy was made according to the Collaborative Review Group search strategy. Data were extracted by two reviewers using the designed extraction form. The software RevMan4.2 was used to review management and data analysis. RESULTS: 67 relevant RCTs were found, of which 12 qualified ones were included into the analysis. Antibiotic prophylaxis significantly decreased the rate of bacteriuria within the period 1 week after TPB (corresponding pooled relative risk (RR) and 95% CI was 0.32 (0.23 - 0.46, P < 0.01). The effective antibiotics included quinolones and quinolones combined with nitroimidazole, with the pooled RR and 95% CI of 0.31 (0.18 - 0.53, P < 0.01) and 0.32 (0.17 - 0.60, P = 0.0004) respectively. There was no significant difference in the effect between short treatment course protocol and long course protocol (P = 0.41). CONCLUSION: Prophylactic antibiotics can significantly decrease the incidence of post-TPB bacteriuria. A significant decrease in bacteriuria incidence can be achieved with a range of antibiotic agents, including quinolones and-quinolones combined with nitroimidazole. Treatment protocols of any duration are effective. PMID- 19552844 TI - [Postprandial changes of blood lipid after ordinary Chinese diet and the influencing factors thereof]. AB - OBJECTIVE: To investigate the postprandial changes of blood lipid after ordinary Chinese diet and the influencing factors thereof. METHODS: Peripheral venous blood samples were collected from 88 patients, 72 males and 16 females, aged (65 +/- 12), 53 with hypertension, 35 with coronary heart diseases, and 27 with diabetes, while fasting and 4 h after breakfast and lunch to measure the levels of lipoproteins, including total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL C), apolipoprotein A (ApoA), ApoB, lipoprotein (Lp) (a), oxidized LDL (ox-LDL). Homeostasis model assessment insulin resistance index (HOMA-IR) was calculated. Weight, height, waist circumference, and hip circumference were examined, and body mass index (BMI) and waist/hip ratio (WHR) were calculated. RESULTS: The TG level after breakfast was (2.79 +/- 0.19) mmol/L, significantly higher than the fasting level by 49.5% [(1.94 +/- 0.13) mmol/L, P < 0.05]. The TG level after lunch was (3.08 +/- 0.26) mmol/L, significantly higher than the fasting level by 58.8% [(1.94 +/- 0.13) mmol/L, P < 0.05]. The ox-LDL after breakfast was (430 +/- 21) microg/L, significantly higher than the fasting level by 32.6% [(324 +/- 14) microg/L, P < 0.05] and the ox-LDL level after lunch was (448 +/- 17) microg/L, significantly higher than the fasting level by 38.1% [(324 +/- 14) microg/L, P < 0.05]. The fasting insulin level was (62 +/- 4) pmol/L, and the HOMA-IR was (15.27 +/- 1.08). The fasting insulin level was positively correlated with the fasting and postprandial TG levels (all P < 0.01). The IR index was positively correlated with the fasting TG and TG after breakfast (both P < 0.05). Fasting insulin and HOMA-IR were negatively correlated with the fasting and postprandial HDL-C levels (all P < 0.05), and positively correlated with the fasting and postprandial TC/HDL-C levels (all P < 0.05). BMI and WHR were negatively correlated with the fasting and postprandial HDL-C (both P < 0.01). BMI was positively correlated with fasting and postprandial TG (both P < 0.05). The insulin level, HOMA-IR, and BMI were negatively correlated with LPL (r = -0.232 - 0.297, P < 0.05). LPL were positively correlated with fasting and postprandial HDL-C levels (r = 0.37, 0.31, 0.35, all P < 0.01). CONCLUSION: The TG and ox-LDL levels significantly increased postprandially. The postprandial blood lipid levels are significantly correlated with the fasting blood lipids. PMID- 19552845 TI - [Multi-channel electrogastrography and autonomic nervous system function in patients with non-erosive reflux disease and reflux esophagitis]. AB - OBJECTIVE: To investigate the gastric myoelectrical activity and autonomic nervous system (ANS) function in patients with non-erosive reflux disease (NERD) and reflux esophagitis (RE). METHODS: 42 patients with gastroesophageal reflux disease (GERD), 21 with NERD and 21 with RE, and 20 healthy volunteers (controls) underwent multi-channel electrogastrography (MEGG) and electrocardiography simultaneously for 30 min in the fasting state and 60 min after a standard test meal. The MEGG parameters included dominant frequency (DF), dominant power (DP), normal percentage of 2 - 4 times/min gastric slow waves (N%), and percentage of slow wave coupling (%SWC). ANS was determined according to power spectra analysis of heart rate variability (HRV) which was conducted by using electrocardiogram recording, including the parameters of sympathetic activity, vagal activity, and sympathovagal balance ratio so as to evaluate the ANS function. RESULTS: The DF levels of the NERD and RE patients and healthy volunteers after test meal were all significantly higher than those in fasting state (all P < 0.05). The %SWC levels of the 3 groups after test meal were all significantly lower than those when fasting (all P < 0.05), and the %SWC level of the NERD patients was significantly lower than that of the healthy volunteers in fasting state [70.5% (40.6% - 81.9%, 76.3% (66.4% - 91.4%), P < 0.05]. Test meal significantly increased the sympathetic activity and sympathovagal balance ratio of the NERD and RE patients and healthy volunteers (all P < 0.05), and the vagal activity levels of the 3 groups after test meal were all significantly lower than those when fasting (all P < 0.01). The sympathovagal balance ratio of the NERD group was significantly lower than that of RE group after meal [1.4 (0.8 - 2.5), 2.2 (1.3 - 4.4), P < 0.05]. CONCLUSION: The spatial regularity of gastric slow waves and ANS function in NERD patients are significantly different from those in RE patients. PMID- 19552846 TI - [Effect of alpha-zearalanol on the generation of reactive oxygen species (ROS) and ROS-activated signal transduction in tumor necrosis factor alpha-stimulated human endothelial cells]. AB - OBJECTIVE: To investigate the effects of alpha-zearalanol (alpha-ZAL) on the generation of reactive oxygen species (ROS) and ROS-activated signal transduction in the tumor necrosis factor (TNF-alpha)-stimulated human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were cultured and divided into 4 groups: (1) normal control group, (2) TNF-alpha stimulated group, undergoing TNF alpha stimulation for 24 h, (3) alpha-ZAL retreatment group, undergoing re treatment with alpha-ZAL of the concentrations of 1 x 10(-8), 1 x 10(-7), or 1 x 10(-6) mol/L for 1 h, then stimulation of TNF-alpha for 24 h, and (4) plasmid transfection group, transfected with p47(phox) siRNA for 24 h to block the NADPH oxidase protein subunit p47(phox) in the HUVECs, or transfected with blank plasmid as control. The intracellular ROS production was detected by using 2, 7 dichlorofluorescin diacetate as probe. Semi-quantitative RT-PCR and immunocytochemistry were used to detect the mRNA and protein expression of p47(phox). The activation of extracellular signal-regulated kinase (ERK) and nuclear translocation of nuclear factor-kappaB (NF-kappaB), stimulatory protein (SP)-1, and activator protein (AP)-1 were assessed with Western blotting. RESULTS: The ROS level in the HUVECs of the TNF-alpha group was higher than that of the control group by 155.4%, and alpha-ZAL reduced the ROS level dose dependently. TNF-alpha treatment up-regulated the p47(phox) mRNA expression by 212.8%, and obviously increased the p47(phox) protein expression; and alpha-ZAL pretreatment attenuated the TNF-alpha-induced p47(phox) mRNA expression by 63.0%, and also markedly inhibited the p47(phox) protein expression. No obvious ROS was found in the HUVECs stimulated by TNF-alpha after the transfection of p47(phox) siRNA. The ERK activation and nuclear translocation of transcription factors SP-1 and NF-kappaB induced by TNF-alpha were abolished or markedly inhibited by alpha ZAL pretreatment. CONCLUSION: alpha-ZAL has a potent inhibitory effect on the ROS production and ROS-activated signaling pathway in the TNF-alpha stimulated endothelial cells, mainly through the inhibition of NADPH oxidase. PMID- 19552847 TI - [Establishment of rat model of venous thromboembolism]. AB - OBJECTIVE: To establish a rat model of venous thromboembolism (VTE). METHODS: One hundred and forty-four SD rats were randomly divided into 2 equal groups: VTE-A group, undergoing local blocking of left femoral artery with micro vessel clip to cause deep vein thrombosis (DVT), and VTE-B group undergoing local blocking of left femoral artery with micro vessel clip in addition of administration of thrombin slowly injected from the distal end of the femoral vein blocked by micro clip. The rate of swelling limbs was observed. One, 4, and 7 days later 6 rats from each group were killed with their femoral veins taken out to observe the thrombosis rate by light microscopy. Other 18 rats in each group underwent injection of the thrombi from left femoral vein solution in normal saline into the right femoral vein 1, 4, and 7 days after DVT formation to establish model of DVT-pulmonary thromboembolism (PTE). The 6 rats of each group [VTE-A-D(n)P(n) and VTE-B-D(n)P(n) groups] were killed 1, 4, and 7 days after DVT-PTE formation respectively with their lungs taken out to observe the rate of PTE by light microscopy. RESULTS: (1) On day 1 after DVT, the successful rate of DVT was 100% in both VTE-A and VTE-B groups, and the swelling limb rate of the VTE-B group was 37.5% (27/72), significantly higher than that of the VTE-A group [16.7% (12/72), P = 0.005]. On day 7 after DVT, the positive thrombus rate of the VTE-B group was 83.3% (20/24), significantly higher than that of the VTE-A group [41.7% (10/24), P = 0.003]. One, 4, and 7 days after DVT, there were reddish, mixed, and organized thrombi in both VTE-A and VTE-B groups. (2) Tachypnea and tachycardia occurred immediately and disappeared spontaneously within 30 - 60 minutes when solution of thrombi was injected into pulmonary arteries via the right femoral veins. One rat died in the VTE-B-D(1)P(1) group 12 h after the embolization and was anatomically confirmed to suffer from fresh massive emboli lodged in pulmonary arteries. The successful rates of DVT-PTE model were 100% (18/18), 83.3% (15/18), and 44.4% (8/18) in the VTE-A-D(1)P(n), VTE-A-D(4)P(n), and VTE-A D(7)P(n) groups respectively, and were 94.4% (17/18), 100% (18/18), and 83.3% (15/18) in the VTE-B-D(1)P(n), VTE-B-D(4)P(n), and VTE-B-D(7)P(n) groups respectively. The successful rate of DVT-PTE model in the VTE-B-D(7)P(n) group was higher than that in the VTE-A-D(7)P(n) group (P = 0.015). The thrombi in pulmonary arteries caused by the 4 or 7 days thrombi of DVT showed lower dissolubility in both VTE-A and VTE-B groups. CONCLUSIONS: A new rat model of VTE (DVT-PTE) has been successfully established. The successful rate of VTE can be increased when thrombin is slowly injected from the distal end of femoral vein blocked by micro clip in addition. PMID- 19552848 TI - [Therapeutic effect of phosphodiesterase-5 inhibitor on pathological changes of tunica albuginea in erectile dysfunction: experiment with rats]. AB - OBJECTIVE: To investigate the pathological changes of tunica albuginea in erectile dysfunction (ED), the role of tunica albuginea in penis erection, and the therapeutic effect of phosphodiesterase-5 inhibitor (PDE5I) on ED. METHODS: Thirty 8-week-old male Sprague-Dawley rats were randomly divided into 3 equal groups: (1) control group fed with normal diet for 16 weeks, (2) high-fat group fed with the diet containing 2% cholesterol and 10% pork fat, and (3) Tadalafil group fed with high-fat diet for 16 weeks and undergoing gastric perfusion of Tadalafil 2 mg/kg daily since the 13th week for 4 weeks. Sixteen weeks later, intracavernous pressure and mean arterial pressure were measured, and the ratio of maximal intracavernous pressure to mean arterial pressure (ICP/MAP) was calculated blood samples were collected. Blood samples were collected from the abdominal aorta to undergo lipid profile test. Then penis was cut. Victoria blue/ponceau red staining and color image analysis were used to observe the content of elastic fibers in the tunica albuginea. RESULTS: The ICP/MAP of the high-fat group was 0.49 +/- 0.05, significantly lower than that of the control group (0.82 +/- 0.06, P < 0.01). The ICP/MAP of the Tadalafil group was 0.60 +/- 0.02, significantly higher than that of the high-fat group (P < 0.05). The content of elastic fibers in tunica albuginea of the high-fat group was 468 +/- 152, significantly lower than that of the the control group (3292 +/- 1123, P < 0.01). The content of elastic fibers in tunica albuginea of the Tadalafil group was 2062 +/- 560, significantly higher than that of the high-fat group (P < 0.05). CONCLUSION: High fat diet may lead to ED and reduction of elastic fibers in tunica albuginea, and PDE-5I reverses such pathological changes. PMID- 19552853 TI - A polio intervention in East African refugees to NSW. AB - This paper summarises a public health intervention in Sydney, NSW in late 2006 that resulted from the potential exposure of a number of refugees to polio virus while in transit in Nairobi, Kenya. The intervention involved the attempted follow-up of 113 persons at risk, assessment for symptoms and immunisation where indicated. No symptomatic cases were found. Seventy-five people were immunised with inactivated poliomyelitis vaccine. The intervention highlighted the importance of close collaboration between health services, the Department of Immigration and Citizenship and settlement service agencies, and provided several lessons to consider when assessing newly arrived refugees. PMID- 19552854 TI - The effectiveness of prophylaxis for measles contacts in NSW. AB - OBJECTIVES: As international estimates of the effectiveness of post-exposure prophylaxis of measles vary, we sought to determine the effectiveness of post exposure prophylaxis with either vaccine or immunoglobulin in susceptible persons with known measles contact. METHODS: Data were obtained on all cases of measles notified in NSW between 1 March and 31 May 2006 and their contacts. The effectiveness of prophylaxis was calculated using the cohort method. RESULTS: During March to May 2006, 57 cases of measles were notified and 1760 measles contacts were identified, of which 553 were classified as susceptible. The calculated effectiveness of post-exposure prophylaxis with vaccine or immunoglobulin in preventing measles was 83.3% (95% CI: 27-96%). CONCLUSION: Post exposure immunisation remains an effective tool for preventing secondary cases of measles. PMID- 19552855 TI - Audit of post-exposure treatment to prevent lyssavirus infection in Sydney South West Area Health Service, 2005-2007. AB - OBJECTIVES: To describe the profile of people who received post-exposure treatment to prevent lyssavirus infection in Sydney South West Area Health Service between 2005 and 2007 and to assess treatment compliance with the current NSW Health protocol. METHODS: Thirty-eight public health files and a subset of 11 medical records were reviewed to collect demographic, exposure and treatment information for the period. RESULTS: Twenty-nine (76%) potential Lyssavirus exposures occurred overseas. Nine potential exposures occurred within Australia; eight of these resulted from a bat bite or scratch. Thirteen (34%) of all potential exposures resulted from handling an animal. CONCLUSION: Many potential exposures were the result of a bite or scratch from a domesticated animal; the animal's survival or health status was not routinely recorded. While all people who commenced post-exposure treatment completed the prescribed course, this was often not within the stipulated timeframe. PMID- 19552856 TI - Another opportunity for prevention: assessing alcohol use by women attending breast screening services in Lismore, NSW. AB - Substantial epidemiological evidence supports the association between alcohol consumption and the risk of breast cancer; however, information on alcohol consumption is not routinely collected by breast screening services in NSW. This study was conducted to investigate the level of self-reported alcohol use among women accessing North Coast Breast Screen in Lismore, NSW. Two hundred and sixty four consecutive women were screened using the Alcohol Use Disorders Identification Test. Their drinking status was categorised as low risk, risky or high risk. Two-thirds of the screened women (69.7%) were classified as low-risk drinkers; 9.8% as risky and 0.4% as high-risk drinkers. Although the risk of breast cancer increases with the amount of alcohol consumed, evidence suggests that even low-risk drinking is associated with increased risk of breast cancer. Implications for prevention activities by breast screening services are discussed. PMID- 19552857 TI - Communicable Diseases Report, NSW, March and April 2009. PMID- 19552858 TI - Helping patients with depression achieve wellness. AB - In the past, response was considered to be the goal of depression treatment. However, studies demonstrated that achieving response was not sufficient for sustaining long-term recovery, so remission (or total elimination of symptoms) became the ultimate aim of treatment. Recently, research has indicated a need to elevate treatment goals from remission to the achieving of wellness-the full restoration of premorbid levels of functioning and quality of life. Strategies such as incorporating objective measures into clinical practice and collaborating with patients throughout therapy will increase the likelihood of attaining wellness. PMID- 19552859 TI - Pharmacotherapy for adult ADHD. AB - The U.S. Food and Drug Administration has approved 3 medications, atomoxetine and the extended-release formulations of amphetamine salts and dexmethylphenidate, for the treatment of adult attention-deficit hyperactivity disorder (ADHD). Different formulations of the same drugs, as well as other agents and cognitive behavioral therapy, have been tested to determine efficacy in ADHD alone and in ADHD with comorbid substance use disorders, mood disorders, and anxiety disorders. A deficit in research exists in regard to these comorbidities in adults with ADHD. PMID- 19552860 TI - Using multidisciplinary care to treat fibromyalgia. AB - Fibromyalgia is a common condition associated with abnormal central pain processing, chronic widespread musculoskeletal pain, tenderness at various points on the body, fatigue, and sleep abnormalities. Individuals with fibromyalgia often have comorbid anxiety, depression, and/or other pain syndromes. Multiple pharmacologic agents have demonstrated efficacy in alleviating some fibromyalgic symptoms, but pharmacology is only one piece of the puzzle in the successful management of fibromyalgia. Thorough medical and psychiatric evaluations are necessary after confirming a diagnosis of fibromyalgia, and a multidisciplinary treatment strategy including patient education, aerobic exercise, and cognitive behavioral therapy as well as pharmacotherapy has been shown to be effective in alleviating fibromyalgic symptoms. PMID- 19552861 TI - Evaluating atypical antipsychotics in the treatment of refractory depression. AB - Treatment resistance is a common clinical occurrence among patients treated for major depressive disorder. Clinicians have several options when treating resistant depression, including switching antidepressants within the same class, switching outside the class, and augmenting with various agents, including atypical antipsychotics. Atypical antipsychotics have shown efficacy as adjuncts to antidepressants in the treatment of resistant depression, although long-term efficacy is undetermined. Atypical antipsychotics need to be compared for efficacy against specific target symptoms, as well as for differences in safety and tolerability. PMID- 19552862 TI - Treating patients with emotional and physical symptoms. AB - Patients with major depressive disorder (MDD) often experience physical pain in addition to depression. Data have shown that as many as 65% of patients with MDD report physical pain. This case vignette presents Ms. S, a 54-year-old nurse with multiple emotional and somatic symptoms, and provides strategies for differentiating and treating the primary and secondary symptoms. For optimal outcomes, emotional and physical symptoms should be treated simultaneously. In addition, a thorough diagnostic process and effective pharmacotherapy are essential to achieving the goal of remission. PMID- 19552863 TI - Norepinephrine and dopamine regulate signals and noise in the prefrontal cortex. AB - ISSUE: Norepinephrine and dopamine work in a cooperative yet reciprocal manner to regulate information processing at pyramidal neurons in the prefrontal cortex. PMID- 19552864 TI - Maintenance of certification in psychiatry. PMID- 19552865 TI - Responses to depressed mood and suicide attempt in young adults with a history of childhood-onset mood disorder. AB - OBJECTIVE: Although individuals' responses to their depressed mood are hypothesized to play an important role in the development and maintenance of depression, how these responses might impact the likelihood of suicidal behavior in mood disorders remains largely unexplored. The goal of the current study was to examine whether maladaptive responses to depressed mood are associated with suicide attempts in adults with a history of childhood-onset mood disorder (COMD). METHOD: Participants included 223 young adult probands with COMD meeting DSM-III or DSM-IV criteria for major depressive disorder or bipolar disorder and 112 controls without a history of psychiatric disorders. All participants were recruited between 1996 and 2004. Probands were followed for 6 to 99 months (median = 32 months). The Responses Styles Questionnaire was used to assess 2 adaptive (distraction and problem solving) and 2 maladaptive (dangerous activity and rumination) ways of coping with depressed mood. RESULTS: Compared to controls, COMD probands scored significantly higher on maladaptive response styles and lower on adaptive styles. Compared to their COMD peers, probands with a history of suicide attempt were less likely to report using distracting activities to manage their depressed mood. However, COMD probands who engaged in dangerous activities in response to depressed mood were more likely to attempt suicide during the follow-up period (hazard ratio = 1.8, 95% CI = 1.2 to 2.8). CONCLUSION: One of the pathways to suicide attempt in mood disorders may involve maladaptive responses to depressed mood. The assessment of how depressed individuals manage their dysphoric moods, therefore, should be considered an important aspect of treatment and prevention of suicidal behavior. PMID- 19552866 TI - A pooled analysis of suicidality in double-blind, placebo-controlled studies of sertraline in adults. AB - OBJECTIVE: The analyses were conducted to identify possibly suicide-related adverse events in Pfizer-sponsored, phases 2 through 4, placebo-controlled, completed studies of sertraline in adult patients and evaluate the risk of suicidality with sertraline versus placebo. METHOD: U.S. Food and Drug Administration (FDA)-defined search methodology was used to identify possibly suicide-related adverse events in short-term, all-duration/all-indication, and psychiatric studies of sertraline. Categorization of possibly suicide-related adverse events was based on the approach developed by the Columbia group for the FDA's analysis of pediatric suicide risk with antidepressants. The incidences of possibly suicide-related adverse events were calculated for individual classifications and for the predefined combined category of suicidality along with the sertraline versus placebo relative risks and corresponding 95% CI limits. Exact binomial CI limits were calculated for the individual treatment group incidences. Age group analyses were also performed using the age limits defined by the FDA. RESULTS: Ninety-nine suicidality events were identified among 19,923 sertraline- and placebo-treated subjects participating in 126 studies conducted between the mid-1980s and the mid-2000s. Four cases of completed suicides among 10,917 sertraline-treated subjects yielded an incidence of 0.04% (95% CI = 0.01 to 0.09) and 3 cases among 9,006 placebo treated subjects yielded an incidence of 0.03% (95% CI = 0.01 to 0.10). There were no statistically significant differences between sertraline and placebo in any of the individual categories or combined suicidality risk category across all performed analyses. CONCLUSION: Results of short-term, all-duration, and psychiatric studies analyses, as well as age-group analyses, performed in accordance with the FDA specified search strategy, show no significant increase in suicidality risk in adult sertraline- versus placebo-treated patients. PMID- 19552867 TI - Duloxetine in the prevention of depressive recurrences: a randomized, double blind, placebo-controlled trial. AB - OBJECTIVE: To assess the efficacy of duloxetine 60-120 mg once daily in the prevention of depressive recurrence in outpatients with recurrent major depressive disorder (MDD). METHOD: Eligible patients with at least 3 episodes of MDD (DSM-IV diagnosis) in the past 5 years received open-label duloxetine 60-120 mg/day for up to 34 weeks. Patients meeting response criteria were then randomly assigned to either duloxetine or placebo for up to 52 weeks of double-blind maintenance treatment. The primary outcome measure was time to recurrence of a major depressive episode. Safety and tolerability were assessed via analysis of treatment-emergent adverse events (TEAEs), vital signs, weight, and laboratory measures. Patients were recruited from 43 study centers in 5 European countries (France, Germany, Italy, Russia, and Sweden) and the United States. The study was conducted from March 2005 to January 2008. RESULTS: A total of 288 patients were randomly assigned to duloxetine or placebo. Time to a depressive recurrence was significantly longer in duloxetine-treated patients compared with placebo-treated patients (p < .001). During the double-blind maintenance phase, 33.1% of placebo treated patients experienced a depressive recurrence compared with 14.4% of duloxetine-treated patients (p < .001). There were no significant differences between treatment groups in TEAEs, discontinuations due to adverse events, vital signs, or weight. CONCLUSIONS: Treatment with duloxetine was associated with a longer time to depressive recurrence and a significantly lower recurrence rate compared with placebo. TRIALS REGISTRATION: (ClinicalTrials.gov) Identifier: NCT00105989. PMID- 19552868 TI - Focus on childhood and adolescent mental health. Predictors, criteria, and treatment response. PMID- 19552869 TI - Suicidal events in the Treatment for Adolescents With Depression Study (TADS). AB - OBJECTIVE: The Treatment for Adolescents with Depression Study (TADS) database was analyzed to determine whether suicidal events (attempts and ideation) occurred early in treatment, could be predicted by severity of depression or other clinical characteristics, and were preceded by clinical deterioration or symptoms of increased irritability, akathisia, sleep disruption, or mania. METHOD: TADS was a 36-week randomized, controlled clinical trial of pharmacologic and psychotherapeutic treatments involving 439 youths with major depressive disorder (DSM-IV criteria). Suicidal events were defined according to the Columbia Classification Algorithm of Suicidal Assessment. Patients were randomly assigned into the study between spring 2000 and summer 2003. RESULTS: Forty-four patients (10.0%) had at least 1 suicidal event (no suicide occurred). Events occurred 0.4 to 31.1 weeks (mean +/- SD = 11.9 +/- 8.2) after starting TADS treatment, with no difference in event timing for patients receiving medication versus those not receiving medication. Severity of self-rated pretreatment suicidal ideation (Suicidal Ideation Questionnaire adapted for adolescents score > or = 31) and depressive symptoms (Reynolds Adolescent Depression Scale score > or = 91) predicted occurrence of suicidal events during treatment (P < .05). Patients with suicidal events were on average still moderately ill prior to the event (mean +/- SD Clinical Global Impressions-Severity of Illness scale score = 4.0 +/- 1.3) and only minimally improved (mean +/- SD Clinical Global Impressions Improvement scale score = 3.2 +/- 1.1). Events were not preceded by increased irritability, akathisia, sleep disturbance, or manic signs. Specific interpersonal stressors were identified in 73% of cases (N = 44). Of the events, 55% (N = 24) resulted in overnight hospitalization. CONCLUSIONS: Most suicidal events occurred in the context of persistent depression and insufficient improvement without evidence of medication-induced behavioral activation as a precursor. Severity of self-rated suicidal ideation and depressive symptoms predicted emergence of suicidality during treatment. Risk for suicidal events did not decrease after the first month of treatment, suggesting the need for careful clinical monitoring for several months after starting treatment. PMID- 19552870 TI - Second-generation antipsychotic-associated diabetes mellitus and diabetic ketoacidosis: mechanisms, predictors, and screening need. PMID- 19552872 TI - Folate supplementation: is it safe and effective? PMID- 19552873 TI - Lupus anticoagulant and anticardiolipin antibodies in serum of patients treated with risperidone. PMID- 19552874 TI - [Bonding in orthodontics. Introduction]. PMID- 19552875 TI - [Methods of bonding to the enamel in orthodontics]. AB - Mastery of the bonding stage of treatment is clearly a critical step in the clinical practice of orthodontics. There is such a wide variety of products available in orthodontics that a reasoned assessment of bonding systems is a practical necessity. Composite plastics, associated with hydrophobic or hydro compatible adhesive systems, and the CVIMAR represent the two principal types of bonding agents used in dentistry. They are categorized according to their constituents into a wide range of products whose nuanced differences are sometimes difficult to discern. This paper first focuses on the development of the composition of the various materials, a depiction of the fundamental parameters of adhesion, and a detailed terminology to help the reader reach a basic understanding. Bonding systems are designed to fulfill the requirements of specific clinical situations. A description of their modes of adhesion, of their composition, and of their advantages and disadvantages will be presented in the second part of this article. PMID- 19552876 TI - [Comparison of 1-stage orthodontic bonding systems and 2-stage bonding systems: a review of the literature and the results of a randomized clinical trial]. AB - OBJECTIVE: The main objectives of this study are to present a literature review of self-etching primer's (SEP's) and present the outcomes of a prospective clinical trial to assess the clinical bond failure rates of orthodontic brackets bonded using a self-etching primer (SEP), compared with brackets bonded using a conventional acid-etched technique with a control adhesive (Transbond). A secondary aim was to investigate whether characteristics of the operator, patient or tooth bonded had any influence on bracket failure. DESIGN: Single-centre randomized controlled clinical trial. Thirty-four patients were bonded, each being randomly assigned to either the test or control adhesive. SETTING: Orthodontic Department Countess of Chester Hospital, Chester, UK. SUBJECTS: Orthodontic patients requiring fixed appliance treatment. MAIN OUTCOME MEASURES: Bond failure. MAIN OUTCOME RESULTS: Failure rates over the initial 6-month period were 2.0% (Transbond) and 1.7% (SEP) with no statistically significant difference between the two groups. Over the duration of the fixed appliance treatment, bond failure rates increased, but remained acceptable at 7.4% (TB) and 7.0% (SEP), respectively. When operator, patient and tooth characteristics were analysed, only the bracket location was found to be significant. Maxillary brackets were more likely to fail than mandibular brackets (relative risk 0.47%; 95% confidence interval 0.22, 1.03). The failure rate for brackets in our study was low when compared with previous studies. CONCLUSIONS: Both the acid-etched control and self-etching primer in combination with adhesive pre-coated brackets were successful for clinical bonding. Their combined failure rate was lower than that reported in similar trials. The literature on SEP's supports the findings of this study. PMID- 19552877 TI - [Enamel micro-abrasion]. AB - Enamel conditioning (elimination of dental plaque and creation of an irregular surface) is an essential step before bonding of orthodontic brackets. The most popular procedure in our practice is bonding with resin which requires enamel etching in order to get enough shear bond strength. Many studies have tried to evaluate the effects of enamel bonding using the acid-etching procedure as well as the changes caused by detachment of brackets. Thanks to the development of other adhesives such as glass ionomer cements which chemically bind to the enamel, new enamel conditioning methods appeared, in particular sandblasting with aluminium oxide particles. This technique is a mechanical preparation of the tooth that avoids the harmful effects of acid products. By suitably choosing the parameters of sandblasting (pressure, time and quantity of powder), enamel loss is lower than with the acid-etch procedure and the surface of the enamel seems less affected. However the bond strength remains superior to the values required for treatment. The presented results indicate that enamel sandblasting can be considered as an alternative for the acid-etching technique currently used in orthodontic practice because it creates sufficient strength and respects enamel thickness better. PMID- 19552878 TI - [Specific bonding technics]. AB - Bonding is a major component of our therapeutic arsenal. We routinely bond attachments to enamel as well as to other surfaces including dentin, ceramic, acrylic, steel, amalgam, and gold, especially for adult patients. This procedure poses certain technical problems for which adapted and specific technical protocols can provide solutions. PMID- 19552879 TI - [Optimal direct bonding methods]. AB - The author explains in this clinical note how the direct bonding method can be most efficiently utilized. He suggests that precision of placement and comfort for patients can be improved if the following two rules are respected: the attachments must be kept horizontal and the substrate must remain immobile. PMID- 19552880 TI - [Methods for bonding Incognito brackets using a chemo-polymerizable bonding agent]. AB - An indirect method is used to bond Incognito attachments. This article will give a step-by-step outline of the procedure that employs a traditional chemo polymerized bonding agent carried in a silicone splint. Using this guide, every practitioner will be able to use a proven technique to achieve the same anticipated results. PMID- 19552881 TI - [Methods for bonding Incognito brackets using a photo-polymerizable bonding agent]. AB - The bonding procedures used in the Incognito system are now well known. Practitioners can work with either a protocol for Maximum-cure which utilizes a hydrophobic and chemo-polymerizable adhesive or an adhesive that is dual and hydrophilic, the association Nexus 3/Excite. Clinically the two protocols provide equivalent results in the quality of the bonding seal for attachments. They are simple to use, but in applying them practitioners must scrupulously follow the manufacturer's recommended stages of procedure. PMID- 19552882 TI - [Indirect bonding technics]. AB - The indirect bonding technique is pivotal for success in lingual orthodontics. There are different laboratory techniques available for indirect positioning and bonding of lingual brackets. Different approaches are presented and described to perform a clinical application. PMID- 19552883 TI - [A simple and quick method for indirect retention]. AB - Retention constitutes a distinct stage of orthodontic therapy. It takes place after the conclusion of active treatment, which makes it a difficult procedure when an orthodontic appliance has been placed on the lingual surfaces of teeth. This article presents a method for fabricating and bonding a retainer during the appointment devoted to removal of the fixed appliance. This technique is a new approach that uses silicone not plaster for obtaining a working model. The step by-step procedure is outlined in detail, from the taking of the impression to the bonding of the retainer. PMID- 19552884 TI - [Bonding and retention]. PMID- 19552885 TI - Engineering of bottlenecks in Rhizopus oryzae lipase production in Pichia pastoris using the nitrogen source-regulated FLD1 promoter. AB - The yeast Pichia pastoris has been previously used for extracellular expression of a Rhizopus oryzae lipase (Rol). However, limitations in Rol folding and secretion through the cell wall became apparent when producing it in fed-batch cultivations. In this study, we have investigated the effect of combining two cell engineering strategies to alleviate putative bottlenecks in Rol secretion, namely the constitutive expression of the induced form of the Saccharomyces cerevisiae unfolded protein response transcriptional factor Hac1 and the deletion of the GAS1 gene encoding beta-1,3-glucanosyltransglycosylase, GPI-anchored to the outer leaflet of the plasma membrane, playing a key role in yeast cell wall assembly. The performance of these engineered Rol-producing strains has been compared in fed-batch cultivations set at a low specific growth rate of about 0.005 h-(1). It was found that Rol overexpression in a P. pastoris strain expressing constitutively the induced form of S. cerevisiae Hac1 and the deletion of GAS1 resulted in about a 3-fold and 4-fold increase in the overall process specific productivity, respectively, whereas the double mutant HAC1/deltagas1 strain yielded about a 7-fold increase. Overall, these results reflect the multiplicity of physiological bottlenecks at different levels/steps throughout the Rol synthesis, secretion and excretion processes in P. pastoris. PMID- 19552886 TI - Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain. AB - Abnormal activity of the epidermal growth factor receptor (EGFR) is associated with various cancer-related processes and motivates the search for strategies that can selectively block EGFR signalling. In this study, functional knockdown of EGFR was achieved through expression of an affibody construct, (ZEGFR:1907)(2 )KDEL, with high affinity for EGFR and extended with the amino acids KDEL to make it resident in the secretory compartments. Expression of (ZEGFR:1907)(2-)KDEL resulted in 80% reduction ofthe cell surface level of EGFR, and fluorescent staining for EGFR and the (ZEGFR:1907)(2-)KDEL construct showed overlapping intracellular localisation. Immunocapture of EGFR from cell lysates showed that an intracellular complex between EGFR and the affibody construct had been formed, further indicating aspecific interaction between the affibody construct and EGFR. Surface depletion of EGFR led to a dramatic decrease in the amount of kinase domain phosphorylated EGFR, coincident with a significant decrease in the proliferation rate. PMID- 19552887 TI - Cellulase production from Aspergillus niger MS82: effect of temperature and pH. AB - Fungal cellulases are well-studied enzymes and are used in various industrial processes. Much of the knowledge of enzymatic depolymerization of cellulosic material has come from Trichoderma cellulase system. Species of Trichoderma can produce substantial amounts of endoglucanase and exoglucanase but very low levels of b-glucosidase. This deficiency necessitates screening of fungi for cellulytic potential. A number of indigenously isolated fungi were screened for cellulytic potential. In the present study, the kinetics of cellulase production from an indigenous strain of Aspergillus niger MS82 is reported. Product formation parameters of endoglucanase and beta-glucosidase (Qp + Y(p/s)) indicate that A.niger MS82 is capable of producing moderate to high levels of both endoglucanase and beta-glucosidase when grown on different carbon containing natural substrates, for example, grass, corncob, bagasse along side purified celluloses. Furthermore, it was observed that the production of endoglucanase reaches its maximum during exponential phase of growth, while b-glucosidase during the Stationary phase. Enzyme production by solid-state fermentation was also investigated and found to be promising.Highest production of cellulase was noted at pH 4.0 at 35 degrees C under submerged conditions. Growth and enzyme production was affected by variations in temperature and pH. PMID- 19552888 TI - Quantification of target proteins using hydrogel antibody arrays and MALDI time of-flight mass spectrometry (A2M2S). AB - Mass spectrometry-based analysis techniques are widely applied in proteomics. This study presents a novel method for quantitative multiplex candidate protein profiling. It applies immunocapture of differentially labeled protein complements on hydrogel antibody arrays and subsequent quantification by MS. To make this approach quantitative a labeling approach was devised. The impact of labeling on the antibody/antigen interaction was assessed in detail by surface plasmon resonance. Owing to there solution by mass more than two protein samples can be compared simultaneously. Direct labeling of crude samples such as sera was developed and so enables the absolute quantification of target proteins straight from crude samples without a protein purification step. It was used to measure the concentration of apolipoprotein A-1 in serum. This method has been termed A2M2S for Affinity Array sand MALDI Mass Spectrometry. PMID- 19552889 TI - Improved microtitre plate production of single chain Fv fragments in Escherichia coli. AB - The new era of functional genomics demands several antibodies as specific detection reagents for proteins, their complexes and post-translational modifications. Only in vitro antibody selection technologies are able to provide the required throughput to generate these large numbers. Phage display is the most widely used technology for in vitro selection of antibodies. The major bottleneck of a phage display selection pipeline is the production of monoclonal antibody fragments for screening and further analysis. In this study, we describe the development of improved protocols for the production of single chain Fv (scFv) antibody fragments in 96-well microtitre plates (MTPs) in Escherichia coli. Four scFvs were expressed using the antibody expression vector pOPE101-XP to analyse the influence of a set of different parameters on their production. Further, six scFvs were expressed using the phage display vector pHAL14 to investigate the effect on the production of functional scFvs using those parameters that improved production from pOPE101-XP. Yield in MTPs was influenced by a variety of conditions and was also strongly dependent on the individual scFv clone. Although it was not possible to deduce a single set of optimal parameters applicable to all the tested scFvs, a combined protocol was developed which improved the expression of scFv fragments over standard methods. PMID- 19552890 TI - The molecular mechanisms of cuticular melanization: the ecdysone cascade leading to dopa decarboxylase expression in Manduca sexta. AB - Many insect developmental color changes are known to be regulated by both ecdysone and juvenile hormone. Yet the molecular mechanisms underlying this regulation have not been well understood. This review highlights the hormonal mechanisms involved in the regulation of two key enzymes [dopa decarboxylase (DDC) and phenoloxidase] necessary for insect cuticular melanization, and the molecular action of 20-hydroxyecdysone on various transcription factors leading to DDC expression at the end of a larval molt in Manduca sexta. In addition, the ecdysone cascade found in M. sexta is compared with that of other organisms. PMID- 19552891 TI - A 25bp-long insertional mutation in the BmVarp gene causes the waxy translucent skin of the silkworm, Bombyx mori. AB - In Bombyx mori, there are more than 35 mutant strains whose larval skin color is transparent. The waxy translucent strain ow is one of the oily mutants which lack accumulation of uric acid in the epidermis. Here we performed positional cloning of the ow gene using the Bombyx draft genome sequence. For fine structure mapping, we succeeded to narrow the ow linked region to approximately 150kb, and identified the ow candidate gene by annotation analysis and DNA sequencing. The complete cDNA sequences of the ow gene from wild-type strains were 3501bp-long and potentially encoded a protein of 920 amino acids. We found a 25bp-long insertion in this gene in the ow mutant strain, resulting in a frame-shift mutation and generation of a premature stop codon. A BLAST search revealed that this protein had high homology to Varp, a recently identified protein containing a vacuolar sorting protein 9 domain and ankyrin repeats, and we termed the silkworm protein BmVarp. Varp has been shown to regulate endosome dynamics, suggesting that BmVarp may play an important role in the incorporation and/or accumulation of uric acid in the epidermis. PMID- 19552892 TI - Cloning and characterization of the Cry1Ac-binding alkaline phosphatase (HvALP) from Heliothis virescens. AB - Membrane-bound alkaline phosphatases (mALPs, EC 3.1.3.1) in the insect midgut have been reported as functional receptors for Cry toxins from the bacterium Bacillus thuringiensis. We previously reported the identification of HvALP in the midgut of Heliothis virescens larvae as a Cry1Ac-binding protein that is down regulated in Cry1Ac-resistant insects. To further characterize HvALP, we localized mALP protein to foregut and midgut tissues using anti-mALP serum and then cloned five mALPs from H. virescens larval midgut. All five clones displayed high levels of sequence identity (above 90%), suggesting that they may represent allelic variants, and grouped with other lepidopteran mALPs in sequence alignments. All these cloned ALPs were predicted to contain a glycosylphosphatidylinositol (GPI) anchor and were named HvmALP1-5. We expressed two of the most diverse HvmALPs in a heterologous system to test binding of Cry1Ac and recognition by HvALP cross-reacting antiserum. Our data highlight the importance of glycosylation for Cry1Ac binding to HvALP and suggest that, depending on glycosylation, all the identified HvmALPs may be synonymous with HvALP, the Cry1Ac-binding phosphatase identified in H. virescens midgut epithelium. PMID- 19552893 TI - The role of NF-kappaB factor REL2 in the Aedes aegypti immune response. AB - Mosquitoes transmit numerous diseases that continue to be an enormous burden on public health worldwide. Transgenic mosquitoes impervious to vector-borne pathogens, in concert with vector control and drug and vaccine development, comprise an arsenal of means anticipated to defeat mosquito-spread diseases in the future. Mosquito transgenesis allows tissue-specific manipulation of their major immune pathways and enhances the ability to study mosquito-pathogen interactions. Here, we report the generation of two independent transgenic strains of Aedes aegypti overexpressing the NF-?B transcriptional factor REL2, a homologue of Drosophila Relish, which is shown to be under the control of the vitellogenin promoter in the mosquito fat body after a blood meal. We show that this REL2 overexpression in the fat body results in transcriptional activation of Defensins A, C, and D, and Cecropins A and N, as well as translation and secretion of Defensin A protein into the hemolymph. We also demonstrate that induction of REL2 results in the increased resistance of the mosquito to tested Gram-negative and Gram-positive bacteria. Importantly, induction of transgenic REL2 leads to the significant decrease in susceptibility of A. aegypti to Plasmodium gallinaceum infection. Consistently, RNAi knockdown of REL2 in wild type mosquitoes results in a delay in Defensin A and Cecropin A expression in response to infection and in increased susceptibility to both bacteria and P. gallinaceum. Moreover, our transgenic assays demonstrate that the N-terminus of the mosquito REL2, which includes the His/Gln-rich and serine-rich regions, plays a role in its transactivation properties. PMID- 19552894 TI - Characterization of the intra-prostatic immune cell infiltration in androgen deprived prostate cancer patients. AB - INTRODUCTION: Our goal was to study the hormonal regulation of immune cell infiltration in prostate cancer patients treated by androgen deprivation therapy (ADT) using an optimized computer-assistance quantification approach. METHODS: The relative density of immune cell subtypes (CD3(+), CD8(+), CD20(+), CD56(+), CD68(+) and Foxp3(+)) was analyzed by immunohistochemistry in archived prostate specimens from control patients (radical prostatectomy only, n=40) and ADT treated patients (ADT prior to radical prostatectomy, n=35) using an image analysis software and a whole-slide scanner. RESULTS: ADT-treated patients had significantly increased relative density of CD3(+) (p<0.001) and CD8(+) T lymphocytes (p<0.001) as well as CD68(+) macrophages (p<0.001). Elevated abundance of CD56(+) Natural Killer (NK) cells was associated with a lower risk of prostate cancer progression (p=0.044), while a high density of CD68(+) macrophages was related to an increased risk of biochemical recurrence (p=0.011). CONCLUSIONS: Our results demonstrate that the infiltration of specific immune cell subtypes is modulated by ADT. Furthermore our data confirm that NK cells have a protective role against tumor progression while macrophages seem to favor the development of advanced prostate cancer. PMID- 19552895 TI - Vaxfectin, a cationic lipid-based adjuvant for protein-based influenza vaccines. AB - Mice were immunized either with unadjuvanted seasonal trivalent influenza vaccine (TIV) or TIV formulated with Vaxfectin, a cationic lipid-based adjuvant. Increasing doses of Vaxfectin resulted in increased hemagglutination-inhibition or anti-TIV ELISA antibody titers, with up to a 200-fold increase obtained with 900 microg of Vaxfectin. A >or=10-fold dose-sparing effect was demonstrated with Vaxfectin formulations. Vaxfectin preferentially increased IgG2 titers compared to IgG1 titers, resulting in a balanced IgG isotype distribution. Lower doses of Vaxfectin (30 microg) did not enhance antibody responses, but increased the number of IFN-gamma secreting T-cells by up to 18-fold. The data demonstrate that Vaxfectin enhances Th1 responses with protein-based seasonal influenza vaccine, and suggest that cellular or humoral immune responses may be preferentially induced by modifying the Vaxfectin:antigen ratio in the vaccine formulation. PMID- 19552896 TI - Glimpses of a one-speed mind: focus-switching and search for verbal and visual, and easy and difficult items in working memory. AB - We investigated focus-switching and search rates in an N-Back task for stimuli presumably encoded either in a phonological/semantic or an abstract-visual format. Experiment 1 used Chinese characters and tested Chinese speakers and non Chinese speakers; character frequency and visual complexity were also manipulated. Experiment 2 presented Chinese characters and English words to non Chinese English speakers. Effects of focus-switching on accuracy were larger for abstract-visual stimuli and for more difficult stimuli; effects on RT were larger for abstract-visual stimuli, but there was no effect of difficulty, with the exception of the most difficult stimulus set in Experiment 1. Search slopes outside the focus of attention did not covary with either type of code or item difficulty. The decline in accuracy over set-size was stronger for the items coded in abstract-visual format. This suggests that item availability is sensitive to robustness of the memory representations, but item accessibility is not. The data fit well with a model of STM in which a fixed number of 'slots' are searched at a constant rate, regardless of the slot's contents. PMID- 19552898 TI - [Urgent intraoperative total enteroscopy with colonoscopy by means of a double enterotomy in a severe lower digestive tract haemorrhage]. PMID- 19552897 TI - Overvaluation of shape and weight in binge eating disorder, bulimia nervosa, and sub-threshold bulimia nervosa. AB - Increasing empirical evidence supports the validity of binge eating disorder (BED), a research diagnosis in the appendix of DSM-IV, and its inclusion as a distinct and formal diagnosis in the DSM-V. A pressing question regarding the specific criteria for BED diagnosis is whether, like bulimia nervosa (BN), it should be characterized by overvaluation of shape and weight. This study compared features of eating disorders in 436 treatment-seeking women comprising four groups: 195 BED participants who overvalue their shape/weight, 129 BED participants with subclinical levels of overvaluation, 61 BN participants, and 51 participants with sub-threshold BN. The BED clinical overvaluation group had significantly higher levels of specific eating disorder psychopathology than the three other groups which did not differ significantly from each other. Findings suggest that overvaluation of shape and weight should not be considered as a required criterion for BED because this would exclude a substantial proportion of BED patients with clinically significant problems. Rather, overvaluation of shape and weight warrants consideration either as a diagnostic specifier or as a dimensional severity rating as it provides important information about severity within BED. PMID- 19552899 TI - Sleep-dependent directional coupling between human neocortex and hippocampus. AB - Complex interactions between neocortex and hippocampus are the neural basis of memory formation. Two-step theories of memory formation suggest that initial encoding of novel information depends on the induction of rapid plasticity within the hippocampus, and is followed by a second sleep-dependent step of memory consolidation. These theories predict information flow from the neocortex into the hippocampus during waking state and in the reverse direction during sleep. However, experimental evidence that interactions between hippocampus and neocortex have a predominant direction which reverses during sleep rely on cross correlation analysis of data from animal experiments and yielded inconsistent results. Here, we investigated directional coupling in intracranial EEG data from human subjects using a phase-modeling approach which is well suited to reveal functional interdependencies in oscillatory data. In general, we observed that the anterior hippocampus predominantly drives nearby and remote brain regions. Surprisingly, however, the influence of neocortical regions on the hippocampus significantly increased during sleep as compared to waking state. These results question the standard model of hippocampal-neocortical interactions and suggest that sleep-dependent consolidation is accomplished by an active retrieval of hippocampal information by the neocortex. PMID- 19552901 TI - Multifactorial analysis of a mixed-longitudinal sample of Indian siblings: Age and sex effects on heritability. AB - The aim of the present study was to examine the heritability of 11 traits in a mixed-longitudinal sample of Indian siblings, and to determine whether heritability estimates vary during the growth period and whether they are influenced by sex. The sample consisted of 245 brothers and 213 sisters from 138 nuclear families living in a semi-urban area in Kolkata, India. The age ranged between 5 and 19 years. The traits were standardised for age and sex using standard deviation scores (SDS) produced by the LMS method (Cole, T.J., 1988. Fitting smoothed centile curves to reference data. J. R. Stat. Soc. A 151, 385 418). The standard deviation scores were analysed by PCA. The two factors with eigenvalues above 1 explained 77.3% of the variance; they showed a high level of pleiotropism present among the studied traits and represented body lengths (PC1) and body weight and breadths (PC2). The heritability between all types of siblings (irrespective of sex) for the PC1 and PC2 was estimated. The heritability between various pairs of siblings showed variations along the whole ontogenetic period studied. During the childhood and pre-pubertal period, heritability between brothers, brother-sister pairs and any sibling pairs was mostly constant, with small and non-significant variations. All the pairs showed the lowest degree of heritability during puberty for PC1 but not for PC2, with significant changes of heritability estimates between adolescence and adulthood, in most of the analysed sibling pairs and in both PC factors. The highest heritability was generally observed at the end of the examined growth period in all pairs. A significant effect of sex on heritability was only detected for PC2 at 11 years of age. PMID- 19552902 TI - Use of uncertainty polytope to describe constraint processes with uncertain time delay for robust model predictive control applications. AB - This paper studies the application of robust model predictive control (MPC) in a constraint process suffering from time-delay uncertainty. The process is described using a transfer function and sampled into a discrete model for computer control design. A polytope is firstly developed to describe the uncertain discrete model due to the process's time-delay uncertainty. Based on the proposed description, a linear matrix inequality (LMI) based MPC algorithm is employed and modified to design a robust controller for such a constraint process. In case studies, the effect of time-delay uncertainty on the control performance of a standard MPC algorithm is investigated, and the proposed description and the modified control algorithm are validated in the temperature control of a typical air-handling unit. PMID- 19552900 TI - Sustained activity within the default mode network during an implicit memory task. AB - Recent neuroimaging studies have shown that several brain regions--namely, the posterior cingulate cortex (PCC), ventral medial prefrontal cortex (vmPFC), and the bilateral angular gyrus--are more active during resting states than during cognitive tasks (i.e., default mode network). Although there is evidence showing that the default mode network is associated with unconscious state, it is unclear whether this network is associated with unconscious processing when normal human subjects perform tasks without awareness. We manipulated the level of conscious processing in normal subjects by asking them to perform an implicit and an explicit memory task, and analyzed signal changes in the default mode network for the stimuli versus baseline in both tasks. The functional magnetic resonance imaging (fMRI) analysis showed that the level of activation in regions within this network during the implicit task was not significantly different from that during the baseline, except in the left angular gyrus and the insula. There was strong deactivation for the explicit task when compared with the implicit task in the default mode regions, except in the left angular gyrus and the left middle temporal gyrus. These data suggest that the activity in the default network is sustained and less disrupted when an implicit memory task is performed, but is suspended when explicit retrieval is required. These results provide evidence that the default mode network is associated with unconscious processing when human subjects perform an implicit memory task. PMID- 19552903 TI - Severe facial rubber bullet injuries: less lethal but extremely harmful weapons. AB - Less-lethal rubber-bullet weapons are designed to induce blunt injuries that incapacitate violent individuals. AIM AND SCOPE: We intend to study the functional and aesthetic impairments and the cost in terms of social health resulting from rubber-bullet facial trauma. MATERIALS AND METHODS: We retrospectively collected all the facial trauma cases caused by mass-appeal, less lethal guns followed up in two French university hospitals since the year 2000. We did not consider the facial injuries caused by professional, less-lethal, rubber or plastic bullet guns. CONCLUSION: We showed that mass-appeal, less lethal rubber-bullet guns induce severe traumas with irreversible functional consequences and long-term social implications. Victims of facial rubber-bullet traumas should be managed like high-energy trauma patients and benefit from extremely careful primary wound care. PMID- 19552904 TI - Short-term metformin treatment for clomiphene citrate-resistant women with polycystic ovary syndrome. AB - OBJECTIVE: To evaluate the effect of a short-course pretreatment with metformin on hyperandrogenism, insulin resistance, cervical scores, and pregnancy rates in women with clomiphene citrate (CC)-resistant polycystic ovary syndrome (PCOS). METHODS: Thirty-seven women with CC-resistant PCOS were randomly assigned to be pretreated with 500 mg of metformin or placebo 3 times per day for 2 cycles, and 100 mg of CC was given on days 5 through 9 of the second cycle in both groups. Luteinizing hormone (LH), follicle stimulating hormone (FSH), dehydroepiandrostendione sulfate (DHEAS), total testosterone (T), glucose, and insulin levels were measured at baseline and after the first cycle, as well as body mass index (BMI), cervical score, and pregnancy rate. RESULTS: After 1 cycle, BMI, total T level, and percentage of participants with insulin resistance were significantly decreased in the metformin group, without any significant decrease in LH, FSH, and DHEAS levels; and in the second cycle, CC treatment resulted in a higher ovulation rate and a thicker endometrium in the metformin group. The pregnancy rate and cervical scores were also higher in that group. CONCLUSION: The short-course pretreatment with metformin decreased hyperandrogenism and insulin resistance and improved cervical sores, ovulation rate, and pregnancy rate among women with CC-resistant PCOS. PMID- 19552905 TI - The effectiveness of interactive computer assisted instruction compared to videotaped instruction for teaching nurses to assess neurological function of stroke patients: a randomized controlled trial. AB - BACKGROUND: Teaching inexperienced nurses to assess neurologic function of acute ischemic stroke patients poses challenges to educators in Taiwan. OBJECTIVES: The purpose of this study was to examine the effectiveness of two programs that teach nurses the use of the Chinese version of the National Institute of Health Stroke Scale (C-NIHSS), and to evaluate the level of learner satisfaction with these teaching programs. DESIGN: An experimental research design with two groups, one pre-test and two post-tests was utilized. SETTING: Six neurology and neurosurgery wards at two hospitals in southern Taiwan. PARTICIPANTS: Participating nurses were stratified based on their clinical level of experience and prior training on the National Institute of Health Stroke Scale (NIHSS). They were randomly assigned to either the experimental C-NIHSS interactive computer assisted instruction (ICAI) group (n=44) or the Instructor-led videotape learning program (IVLP) group (n=40) to learn the C-NIHSS. METHODS: The measurement tools included the score verification unit (SVU) (score range from 0 to 45, content validity index, CVI=0.96, percentage agreement=84%) and the learner satisfaction scale (CVI=0.92, Cronbach's alpha=0.97). RESULTS: Both groups' scores on the assessment of correctness significantly increased (F=35.50, p=0.00) after intervention. However, there was an insignificant difference between the changes in the two groups (F=0.02, p=0.89). After using one-way ANCOVA analysis, and adjusting for the length of experience in neurological nursing, the results showed that in the second post-test, the ICAI group's score was significantly higher than that of the IVLP group (F=4.81, p=0.03). There was a positive correlation between assessment correctness on the second post-test and length of experience in neurological nursing (r=0.35, p<0.05). It was concluded that nurses with less experience in neurological nursing, who receive ICAI will perform a better assessment of stroke patients than those who received IVLP. CONCLUSION: The C NIHSS ICAI teaching program contributed to better assessment correctness after adjusting for the length of experience in neurological nursing, and to some extent increased satisfaction for the participating nurses. Therefore it is worth promoting the use of ICAI for in-service education of nurses, especially for nurses with less experience in neurological nursing, in order to enhance long term effects of learning. PMID- 19552906 TI - [Appendiceal mucoceles, pseudomyxoma peritonei and appendiceal mucinous neoplasms: update on the contribution of imaging to choice of surgical approach]. AB - INTRODUCTION: The treatment of pseudomyxoma peritonei (PMPs) and appendiceal mucocele (AM) has changed radically. To assess the contribution of preoperative imaging to the treatment strategy and choice of approach, a surgeon and a radiologist different from the initial radiologist examined the files of all patients treated for PMP or AM in four facilities in one district from January 1, 1996, through December 31, 2008. PATIENTS AND METHODS: The study included 27 patients (20 men and seven women, mean age: 63+/-13 years). Eleven patients had an intact AM, seven synchronous PMP (malignant appendiceal lesion in two of seven), six metachronous PMP (five with peritoneal mucinous carcinomatosis and one with diffuse peritoneal adenomucinosis) and three a ruptured AM but not PMP. The incidence of mucin-secreting tumors observed (27 cases in 12 years in a region of 500 000 inhabitants) corresponds to a prevalence of approximately five cases per year per million inhabitants. Acute clinical pictures (7/27) were significantly more frequent for the malignant forms (5/7) (p<0,02). RESULTS AND DISCUSSION: The overall sensitivity of computed tomography (CT) for all the criteria studied was 93%. The predictive value for AM rupture of visualization of thick calcifications was 100%. On the other hand, rupture never occurred when the CT showed an AM under pressure, with thin walls and septa. The predictive value for PMP of "scalloping" was 100%. The diagnostic accuracy of the initial reading was 25/27 for the imaging overall and 25/25 for the CT. Preoperative visualization of the exact size of the intact AM or of diagnostic information about ruptured AM and PMP helped to select an appropriate approach in 25 of 27 cases. PMID- 19552907 TI - A new insight into resveratrol as an atheroprotective compound: inhibition of lipid peroxidation and enhancement of cholesterol efflux. AB - Resveratrol, a polyphenolic constituent of red wine, is known for its anti atherogenic properties and is thought to be beneficial in reducing the incidence of cardiovascular diseases (CVD). However, the mechanism of action by which it exerts its anti-atherogenic effect remains unclear. In this study, we investigated the relationship between the antioxidant effects of resveratrol and its ability to promote cholesterol efflux. We measured the formation of conjugated dienes and the rate of lipid peroxidation, and observed that resveratrol inhibited copper- and irradiation-induced LDL and HDL oxidation as observed by a reduction in oxidation rate and an increase in the lag phase (p<0.05). We used DPPH screening to measure free radical scavenging activity and observed that resveratrol (0-50microM) significantly reduced the content of free radicals (p<0.001). Respect to its effect on cholesterol homeostasis, resveratrol also enhanced apoA-1-mediated cholesterol efflux (r(2)=0.907, p<0.05, linear regression) by up-regulating ABCA-1 receptors, and reduced cholesterol influx or uptake in J774 macrophages (r(2)=0.89, p<0.05, linear regression). Incubation of macrophages (J774, THP-1 and MPM) with Fe/ascorbate ion, attenuated apoA-1 and HDL(3)-mediated cholesterol efflux whereas resveratrol (0-25microM) significantly redressed this attenuation in a dose-dependent manner (p<0.001). Resveratrol thus appears to be a natural antioxidant that enhances cholesterol efflux. These properties make it a potential natural antioxidant that could be used to prevent and treat CVD. PMID- 19552909 TI - Molecular-shape selectivity tuned by donor-acceptor type copolymers as organic phase in reversed-phase high-performance liquid chromatography. AB - The copolymer with a terminal reactive group (copoly-VP17MA12) was newly designed and prepared by selecting 4-vinylpyridine (VP) as an electron-donating monomer and methyl acrylate (MA) as an electron-accepting monomer. Copoly-VP17MA12 was grafted onto porous silica particles by a silanol coupling reaction with the terminal group. Individually, poly-VP15 and poly-MA20 with a terminal reactive group were also synthesized and co-grafted onto silica (Sil-cograft-VP17MA12) to compare the effect of the copolymer-grafted (Sil-copoly-VP17MA12) and polymer co grafted stationary phases in HPLC separation. All the polymers were characterized by 1H NMR and FT-IR analysis before grafting on silica. Polymer grafting was confirmed by diffuse reflectance infrared Fourier transform (DRIFT), elemental analysis and thermogravimetric analysis (TGA). The intra- and intermolecular interaction between monomer units in copolymer-grafted and polymer co-grafted systems, respectively was observed by DRIFT measurement. The detailed chromatographic study revealed using PAHs as solutes that remarkably higher selectivity for planar/non-planar discrimination was observed with the copolymer grafted phase than with the co-grafted stationary phase. In this paper, it is also discussed that the selectivity enhancement by copolymer-grafting in the Sil copoly-VP17MA12 phase can be brought through intramolecular interaction between VP and MA. PMID- 19552908 TI - Synthesis of dinitrophenyl-L-Pro-N-hydroxysuccinimide ester and four new variants of Sanger's reagent having chiral amines and their application for enantioresolution of mexiletine using reversed-phase high-performance liquid chromatography. AB - Four chiral derivatizing reagents (CDRs) having enantiomerically pure amines and two CDRs namely, [N-succinimidyl-(S)-2-(6-methoxynaphth-2-yl)propionate], and [dinitrophenyl-L-Pro-N-hydroxysuccinimide ester, DNP-L-Pro-SU] were synthesized and were used to prepare diastereomers of (R,S)-mexiletine (MEX); these were separated by reversed-phase high-performance liquid chromatography (RP-HPLC). The method was validated for linearity, accuracy, limit of detection (LOD) and limit of quantification (LOQ). PMID- 19552910 TI - Simultaneous determination of fluoroquinolones, sulfonamides and tetracyclines in sewage sludge by pressurized liquid extraction and liquid chromatography electrospray ionization-mass spectrometry. AB - A new scheme for the quantitative determination of traces of fluoroquinolones (FQs), tetracyclines (TCs) and sulfonamides (SAs) in sewage sludge was developed. The compounds were simultaneously extracted from sewage sludge by pressurized liquid extraction (PLE). A novel and effective method for PLE was developed. Solid-phase extraction was used for cleaning up the extracts. Identification and quantification of the compounds was done using high-performance liquid chromatography with electrospray ionization mass spectrometry in selected reaction monitoring mode. The best recovery of FQs and TCs was obtained by using hydrophilic-lipophilic balance cartridges, recoveries ranged 59% for norfloxacin to 82% for ofloxacin and 95% for doxycycline; for SAs strong cation-exchange cartridges were more efficient, recoveries were 96% for sulfamethoxazole and 43% for sulfadimethoxine. Limit of quantification ranged from 0.1 ng/g for SAs to 160 ng/g for tetracycline. Method precision for TCs was 5.06% and 1.12%, and for SAs 0.43% and 2.01%. FQs precision ranged from 0.77% to 1.89%. PMID- 19552911 TI - Enantioresolution of basic pharmaceuticals using cellulose tris(4-chloro-3 methylphenylcarbamate) as chiral stationary phase and polar organic mobile phases. AB - A polysaccharide-based chiral stationary phase (Sepapak-4), with cellulose tris(4 chloro-3-methylphenylcarbamate) as chiral selector, has been investigated in liquid chromatography (LC). Its enantioresolution power was evaluated towards 13 basic amino-drugs with widely different structures and polarities, using polar organic mobile phases. After preliminary experiments, acetonitrile was selected as the main mobile phase component, to which a low concentration of diethylamine (0.1%) was systematically added in order to obtain efficient and symmetrical peaks. Different organic solvents were first added in small proportions (5-10%) to acetonitrile to modulate analyte retention. Polar organic modifiers were found to decrease retention and enantioresolution while hexane had the opposite effect, indicating normal-phase behaviour under these conditions. The addition of an organic acid (formic, acetic or trifluoroacetic acid) was found to strongly influence the retention of the basic amino drugs in these nonaqueous systems. The nature and proportion of the acidic additive in the mobile phase had also deep impact on enantioresolution. Therefore, the studied compounds could be subdivided in three groups in respect to the acidic additive used. All analytes could be enantioseparated in relatively short analysis times (10-20 min) using these LC conditions. PMID- 19552912 TI - On the retention mechanisms and secondary effects in microthermal field-flow fractionation of particles. AB - The behavior of nanometer or micrometer-sized particles, dispersed in liquid phase and exposed to temperature gradient, is a complex and not yet well understood phenomenon. Thermal field-flow fractionation (TFFF), using conventional-size channels, played an important role in the studies of this phenomenon. In addition to thermal diffusion (thermophoresis) and molecular diffusion or Brownian movement, several secondary effects such as particle particle and/or particle-wall interactions, chemical equilibria with the components of the carrier liquid, buoyant and lift forces, etc., may contribute to the retention and complicate the understanding of the relations between the thermal diffusion and the characteristics of the retained particles. Microthermal FFF is a new high-performance technique allowing much easier manipulation and control of the operational parameters within an extended range of experimental conditions in comparison with conventional TFFF. Consequently, in combination with various other methods, it is well suited for a detailed investigation of the mentioned effects. In this work, some contradictory published results concerning the thermal diffusion of the colloidal particles, studied by TFFF but also by other methods, are analyzed and compared with our experimental findings. PMID- 19552913 TI - A facile dip-coating process for preparing highly durable superhydrophobic surface with multi-scale structures on paint films. AB - Superhydrophobic surfaces with multi-scale nano/microstructures have been prepared on epoxy paint surfaces using a feasible dip-coating process. The microstructures with 5-10 microm protuberances were first prepared on epoxy paint surface by sandblast. Then the nanostructures were introduced on the microstructure surface by anchoring 50-100 nm SiO(2) particles (nano-SiO(2)) onto the sandblasted paint surface, which was completed by dip-coating with a nano SiO(2)/epoxy adhesive solution (M1). At last the surface was further modified for enhancing hydrophobicity by another dip-coating with a solution of a low surface energy polymer, aminopropyl terminated polydimethylsiloxane (ATPS) modified epoxy adhesive (M2). The water contact angle of the as-prepared samples reached as high as 167.8 degrees and the sliding angle was 7 degrees. The prepared superhydrophobic surface exhibited excellent durability to the high speed scouring test and high stability in neutral and basic aqueous solutions and some common organic solvents. In addition, this method can be adopted to fabricate large scale samples with a good homogeneity of the whole surface at very low cost. PMID- 19552914 TI - The effect of surface charge distribution on hindered diffusion in pores. AB - An analytic solution is derived for the hindered diffusion of charged, small solutes in charged, cylindrical pores in which the pore wall potential consists of the sum of an average and an oscillatory component. When the oscillatory contribution is absent, the effect of electrostatic interactions on diffusion is negligible. However, when the wall potential or surface charge density varies axially, electrostatic interactions hinder the rate of diffusion significantly, and can stop it completely if "choke points" develop where the solute concentration becomes zero. The degree of hindrance is generally weaker when the electrostatic charge on the pore wall and the charge on the solute have the same signs, leading to a repulsion, than it is in the presence of an attraction. The electrostatic hindrance is also affected by the length scale of the axial variation along the pore wall, becoming stronger as that length grows, until an asymptotic value is reached. The theory for the effect of variations of the electrostatic potential on rates of diffusion is shown to be in good agreement with experimental data taken from the literature. The results here are obtained by using generalized Taylor dispersion theory, and are therefore rigorous predictions of what occurs over times long enough that the solute diffuses through a tube many times longer than a single periodic cell. The electrostatic interactions are calculated using the linear Poisson-Boltzmann equation. PMID- 19552915 TI - Synthesis and characterization of Prussian blue@platinum nanoparticle hybrids from a mixture solution of platinum nanocatalyst and ferric ferricyanide. AB - In the present study, platinum nanoparticles modified with Prussian blue (PB) have been synthesized by a heterogeneous catalytic reaction. Transmission electronic microscopy (TEM) confirmed the deposition of nanoclusters around the surfaces of platinum particles, and spectroscopic studies verified that the molecular composition of the nanoclusters was dominantly PB and a minority of platinum ferricyanide. Thus, it was shown that the platinum particles behaved not only as catalysts for the growth of PB, but also as a reactant to generate a PB analogue complex. Moreover, potassium was not detected in the final product, demonstrating that the molecular nature of PB was in the "water-insoluble" form, Fe(4)(3+)[Fe(II)(CN)(6)](3). PMID- 19552916 TI - Determination of diffusion and sorption parameters of thin confined clay layers by direct fitting of through-diffusion flux. AB - Diffusion in compacted clays is often studied in sandwich-like arrangements where the clay is confined by porous filter plates in order to control its swelling. In some clays (for example, Na-montmorillonite) equilibrated with dilute electrolyte solutions, the fluxes of cationic radiotracers can be quite high due to cation exchange reactions. Accordingly, the diffusion resistance of clay layers can become comparable with or even smaller than the diffusion resistance of porous filters (such layers are called "thin" in this study). In view of the typical uncertainties (ca. 20%) of diffusion permeability of porous filters reported in the literature, the diffusion resistance of clay layers cannot be reliably determined from the steady-state diffusion permeability of the filter-clay-filter "sandwich" in this case. In this study, it is shown that, rather unexpectedly, information on the diffusion permeability of "thin" clay layers can be obtained from the time dependence of diffusant flux into the outlet compartment because at very short times, there is a characteristic flux delay that does not occur in the limiting case of infinitely large diffusion permeability of clay. The flux behavior at longer times is controlled by the diffusion permeability of the filters, which makes possible its determination directly from through-diffusion data and makes superfluous independent diffusion experiments with filters. This approach has been validated via theoretical interpretation of literature data on the diffusion of (22)Na radiotracers through confined compacted montmorillonite equilibrated with 0.01 M NaClO(4) solution. The filter and clay properties estimated in this way are in good agreement with the literature data. PMID- 19552917 TI - Synthesising gold nanoparticles within bola fatty acid nanosomes. AB - We report on the synthesis of gold nanoparticles (Au-NP) stabilized by a layer of mercapto and/or hydroxyl bola palmitic acid-tetrabutyl ammonium salts (TBAOH) which form nanosomes in water. Mixing both bola fatty acids in water in various proportions allowed to tune the nanoparticle diameter from 1 to 10 nm using NaBH(4) as a reductor. Those nanoparticles could be phase transferred to dichloromethane by adding a more hydrophobic counter-ion than TBAOH, its derivative with octyl chains (TOAOH). Finally, we used those Au-NP to formulate emulsions, the stability of which was followed as a function of particle size and time. The emulsions were stable but creamed after several hours. We conclude that bola fatty acid nanosomes are then good candidates to synthesize gold nanoparticles dispersed either in water, organic solvents and emulsions. PMID- 19552918 TI - The role of higher level adaptive coding mechanisms in the development of face recognition. AB - Developmental improvements in face identity recognition ability are widely documented, but the source of children's immaturity in face recognition remains unclear. Differences in the way in which children and adults visually represent faces might underlie immaturities in face recognition. Recent evidence of a face identity aftereffect (FIAE), in which adaptation (exposure) to a particular identity causes a previously neutral face to take on the computationally opposite identity, suggests that adults code faces in an opponent fashion relative to an average face. One previous study showed comparable FIAEs in 8-year-olds and adults but did not demonstrate that adaptation was selective for high-level representations in both groups. Using a developmentally appropriate FIAE task, we investigated whether children show adult-like adaptation for facial identity when adapting and test images differ in size. Both age groups showed an equivalent FIAE, suggesting that qualitative changes in the use of higher level adaptive coding mechanisms do not drive the developmental improvements in face recognition ability, at least from 8 years of age. PMID- 19552919 TI - Low serum IL-10 concentrations and loss of regulatory association between IL-6 and IL-10 in adults with major depression. AB - Elevated circulating pro-inflammatory cytokines are associated with symptoms of depression, and disorders involving chronic inflammation are often co-morbid with major depression. Since healthy immune regulation is accomplished through counter balancing effects of pro- and anti-inflammatory cytokines, we hypothesized that depressed subjects (compared to controls) would express lower concentrations of the anti-inflammatory/immunoregulatory cytokine interleukin (IL)-10, and a higher IL-6/IL-10 ratio. We also examined the possibility that depressed subjects may exhibit a deficiency in the regulatory loop involving IL-6 induced secretion of IL-10. Therefore, we hypothesized that circulating IL-6 and IL-10 would be positively correlated in controls, while the correlation would be weaker in depressed subjects. Resting state serum cytokine concentrations were quantified in 12 unmedicated depressed subjects, and 11 age, gender, and ethnicity-matched controls. Depressed subjects showed significantly lower IL-10 (p=0.03, Cohen's d= 0.96), non-significantly higher IL-6, and significantly higher IL-6/IL-10 ratios (p=0.05, Cohen's d=0.50). Across all participants, higher scores on the self rated Inventory of Depressive Symptoms were associated with lower IL-10 (r(21)= 0.57, p=0.005) and non-significantly higher IL-6/IL-10 ratios (r(21)=0.38, p=0.07), but not related to IL-6 concentrations. As hypothesized, IL-6 and IL-10 concentrations were strongly and positively correlated in controls (r(9)=0.81, p=0.003), but were completely dissociated in depressed subjects (r(10)=0.01, p=0.98). These results suggest that lower IL-10 levels, a higher IL-6/IL-10 ratio, and the apparent absence of a counter-balancing, immunoregulatory increase in IL-10 in response to elevated IL-6 concentrations contribute to the pro inflammatory physiological milieu that is known to be associated with major depression. Therefore, reduced induction/availability of IL-10, that would normally inhibit pro-inflammatory cytokine actions and resolve inflammation, may contribute to the depressogenic as well as the inflammatory disease-promoting effects of chronic, low-level elevations in pro-inflammatory cytokines. PMID- 19552920 TI - Is NOS1 a genetic link between RLS and ADHD? PMID- 19552921 TI - Cardiac surgery in patients with major lower extremity amputation: a single institution experience. AB - BACKGROUND: Cardiac surgery patients with lower-extremity amputations pose a challenge in terms of medical comorbidities and functional recovery. METHODS: A retrospective review of all patients (n=10) with preexisting below-knee amputation (BKA) or more proximal amputation level who underwent cardiac surgery between April 1998 and April 2008. Data were analyzed to evaluate outcomes. RESULTS: The median age was 59 y (range, 51-75 y). One patient had bilateral above-knee amputation (AKA), and 9 had BKAs (two bilateral). Comorbidities included diabetes (n=5), peripheral vascular disease (n=7), cerebrovascular disease (n=2), hypertension (n=9), chronic renal insufficiency (n=2), pulmonary hypertension (n=1), and pulmonary fibrosis (n=1). Nine patients underwent coronary artery bypass grafting and one patient underwent aortic valve replacement. There were no operative deaths. The median length of hospital stay (to home discharge) was 12.5 d (range, 5-562 d). Eight patients were transferred to a rehabilitation unit or a chronic care facility before being discharged to home. At follow-up (median, 1.5 y; range, 0.4-3.8 y), all but one patient were alive and had returned to their preoperative ambulatory status. CONCLUSIONS: In our experience, patients with lower-extremity amputations require prolonged hospitalization after cardiac surgery but can expect good mid-term outcomes and functional recovery. PMID- 19552922 TI - Meta-analysis comparing arthroplasty with internal fixation for displaced femoral neck fracture in the elderly. AB - BACKGROUND: The treatment of displaced femoral neck fracture includes internal fixation and arthroplasty. However, which is the best surgical treatment for the elderly patient with displaced femoral neck fractures has been controversial. Our objective was to compare the clinical effects of internal fixation with that of arthroplasty for displaced femoral neck fracture in the elderly (>=60 y of age). MATERIALS AND METHODS: We searched for all randomized controlled trials of hip arthroplasty versus internal fixation for displaced femoral neck fractures in the elderly by electronically searching PUBMED (1966 to December, 2008), MEDILINE (1966 to December, 2008) and manually searching grey literatures. The quality of the trials was assessed and meta-analyses were conducted using the Cochrane Collaboration's RevMan 4.2 software. RESULTS: Nineteen published randomized controlled trials involving a total of 3505 patients were suitable for inclusion in the review. The combined results of meta-analyses showed no significant difference in mortality at 1 y postoperatively between the two methods. However, compared with internal fixation, arthroplasty could reduce the rate of reoperations and the major method-related complications. CONCLUSIONS: Compared with internal fixation, arthroplasty can not only reduce the surgical revision, but also decrease the incidence of complications, and does not increase mortality. The present meta-analysis shows that there is an evidence base to support arthroplasty as a primary treatment for displaced femoral neck fractures in the elderly. PMID- 19552923 TI - Single dose GLP-1-Tf ameliorates myocardial ischemia/reperfusion injury. AB - BACKGROUND: Glucagon-like peptide-1 (GLP-1) has insulinomimetic, insulinotropic, and antiapoptotic properties that may make it a useful adjunct to reperfusion therapy for myocardial infarction (MI); however, GLP-1 has a short plasma half life. Fusion of GLP-1 to human transferrin (GLP-1-Tf) significantly prolongs drug half-life. MATERIALS AND METHODS: We tested the ability of single dose GLP-1-Tf to limit myocardial ischemia (30 min)/reperfusion (180 min) injury in rabbits. Nineteen animals were untreated controls. The pre-ischemic group (n=10) was given 10mg/kg of GLP-1-Tf 12 h before ischemia. Immediately after reperfusion, the post ischemic group (n=10) received GLP-1-Tf (10 mg/kg) and the Tf group (n=4) received transferrin alone. RESULTS: Infarct size as a percentage of the area at risk was 59.1% +/- 1.3%, 45.7% +/- 1.9%, 44.1% +/- 3.3%, 59.7% +/- 2.0% in the control group, pre-ischemic group, post-ischemic group, and Tf group, respectively (P<0.05 for both GLP-1-Tf treatments group versus control). GLP-1-Tf reduced the apoptotic index from 4.67% +/- 0.40% in the control group to 3.15% +/ 0.46% in the pre-ischemic group and to 2.66% +/- 0.40% in the post-ischemic group (P<0.05 for both GLP-1-Tf treatments versus control). The size of the wall motion abnormality and ejection fraction was significantly improved in the post ischemic group relative to the control group. Serum GLP-1 levels were 239.8 +/- 25.7 MUg/mL in the post-ischemic group, 27.9 +/- 5.8 MUg/mL in the pre-ischemic group, and undetectable in the control group. CONCLUSION: GLP-1-Tf limits myocardial reperfusion injury whether given prior to the onset of ischemia or given at reperfusion. GLP-1-Tf may also limit myocardial stunning at high serum levels of the drug. PMID- 19552925 TI - Low uric acid levels in serum of patients with ALS: further evidence for oxidative stress? AB - BACKGROUND: The exact cause of amyotrophic lateral sclerosis (ALS) is unknown. Oxidative stress is one of the factors implicated in the etiology of ALS as well as in that of other neurodegenerative diseases. Uric acid is an important natural antioxidant that may reduce oxidative stress. The objective of this study was to prospectively determine the serum uric acid levels in ALS patients and allegedly healthy individuals and to correlate those values with measures of ALS disease progression among the patients. METHODS: The ALS patients and well-matched controls underwent blood tests for serum uric acid levels which were then correlated with the patients' disability status, as expressed by the ALS Functional Rating Scale (ALSFRS-R). RESULTS: Eighty-six ALS patients and 86 well matched controls participated. The ALS patients' mean+/-SD uric acid level was significantly lower (4.78+/-1.3 mg/dl) than that of the controls (5.76+/-1.26 mg/dl) (p<0.0001). The findings were similar for a second examination performed after an interval of at least 6 months. There was a correlation between the relative decrease of serum uric acid levels among patients (the difference between the patients' level and the controls' level) and the rate of disease progression (ALSFRS-R decline) (p<0.0001, r=0.624). CONCLUSIONS: ALS patients had lower serum uric acid levels than healthy individuals. The decreased uric acid levels were correlated to the rate of disease progression (ALSFRS-R decline), further demonstrating the possible role of oxidative stress in the induction and propagation of the disease. PMID- 19552924 TI - Follicular thyroid cancer cell growth inhibition by proteosome inhibitor MG132. AB - BACKGROUND: Effective therapies for the subset of follicular thyroid cancer (FTC) patients with aggressive, metastatic disease are lacking. Therefore, we sought to determine the effects of proteosome inhibition, an emerging class of chemotherapeutic agents, on metastatic FTC cells. MATERIALS AND METHODS: Human metastatic FTC cells (FTC236) were treated in vitro with the proteosome inhibitor MG132 (0 to 800 nM). Western blot analysis was performed on whole cell lysates isolated after 2 d. To measure cell growth, we performed an MTT cellular proliferation assay over 6 d. RESULTS: Treatment of FTC236 cells with MG132 led to dose-dependent cell growth inhibition. Increases in inactive, phosphorylated GSK-3beta, and active beta-catenin also were observed. With 800 nM MG132, growth was reduced by 87% at 6 d (P < 0.0001). This reduction in cellular proliferation correlated with the degree of GSK-3beta inhibition. MG132 treatment also caused increased p21(Waf1/Cip1) and decreased cyclin D1 expression, suggesting that growth suppression may occur through cell cycle arrest. CONCLUSION: Growth of metastatic human FTC cells appears to be suppressed by proteosome inhibition. Whether this effect is directly due to cell cycle arrest and inactivation of GSK 3beta signaling is unclear. Nonetheless, these compounds may become novel treatments for aggressive, metastatic FTC. PMID- 19552926 TI - Comparison study of amyloid PET and voxel-based morphometry analysis in mild cognitive impairment and Alzheimer's disease. AB - Two techniques employed for the early diagnosis of dementia are the imaging of amyloid-beta protein using positron emission tomography (PET) and voxel-based morphometry analysis of MRI (VBM-MRI). The purpose of this study was to evaluate the clinical utility of amyloid PET and VBM-MRI for the early diagnosis and tracking of the severity of Alzheimer's disease (AD). The neuritic plaque burden and gray matter losses were evaluated using [11C]BF-227-PET and VBM-MRI in 12 healthy controls, 13 subjects with mild cognitive impairment (MCI), including 6 who converted to AD and 7 who did not convert, and 15 AD patients. The AD patients and the MCI converters exhibited a neocortical retention of BF-227 and parahippocampal gray matter loss shown by VBM-MRI. The MCI converters were more clearly distinguished from the MCI non-converters in BF-227-PET than VBM-MRI. The combined sample of the MCI converters and AD patients showed a significant correlation of MMSE scores with the global gray matter loss, but not with the BF 227 retention. These findings suggest that amyloid PET using [11C]BF-227 is better suited for the prediction of conversion from MCI to AD, while VBM-MRI appears to be better suited for tracking the severity of dementia. PMID- 19552928 TI - [Intralobar pulmonary sequestration]. PMID- 19552927 TI - Increased intra-individual reaction time variability in attention deficit/hyperactivity disorder across response inhibition tasks with different cognitive demands. AB - One of the most consistent findings in children with ADHD is increased moment-to moment variability in reaction time (RT). The source of increased RT variability can be examined using ex-Gaussian analyses that divide variability into normal and exponential components and Fast Fourier transform (FFT) that allow for detailed examination of the frequency of responses in the exponential distribution. Prior studies of ADHD using these methods have produced variable results, potentially related to differences in task demand. The present study sought to examine the profile of RT variability in ADHD using two Go/No-go tasks with differing levels of cognitive demand. A total of 140 children (57 with ADHD and 83 typically developing controls), ages 8-13 years, completed both a "simple" Go/No-go task and a more "complex" Go/No-go task with increased working memory load. Repeated measures ANOVA of ex-Gaussian functions revealed for both tasks children with ADHD demonstrated increased variability in both the normal/Gaussian (significantly elevated sigma) and the exponential (significantly elevated tau) components. In contrast, FFT analysis of the exponential component revealed a significant task x diagnosis interaction, such that infrequent slow responses in ADHD differed depending on task demand (i.e., for the simple task, increased power in the 0.027-0.074 Hz frequency band; for the complex task, decreased power in the 0.074-0.202 Hz band). The ex-Gaussian findings revealing increased variability in both the normal (sigma) and exponential (tau) components for the ADHD group, suggest that both impaired response preparation and infrequent "lapses in attention" contribute to increased variability in ADHD. FFT analyses reveal that the periodicity of intermittent lapses of attention in ADHD varies with task demand. The findings provide further support for intra-individual variability as a candidate intermediate endophenotype of ADHD. PMID- 19552929 TI - [The role of diffusion-weighted imaging in the evaluation of meningiomas: radio pathologic correlation]. AB - OBJECTIVE: To study the behavior of meningiomas in diffusion-weighted sequences and the correlation of these findings with the histological findings. MATERIAL AND METHODS: We prospectively included all patients operated on for meningiomas at our hospital during two years. We studied 30 meningiomas in 28 patients aged 31 to 85 years old. All patients underwent MRI prior to surgery, including diffusion-weighted sequences, in a 1.5 T scanner. We evaluated the signal intensity in T2-weighted images, diffusion-weighted images (b=1,000), and apparent diffusion coefficient (ADC) maps within the tumor and in the parietal white matter as a reference. In the histological study, cellularity, proliferation index, histological grade, and cerebral invasion were evaluated. RESULTS: Of the 30 meningiomas, 22 were World Health Organization (WHO) grade I and 8 were atypical or WHO grade II. The overall mean value of the ADC was 89.19+/-13.95x10(-3) mm2/s; the mean ADC value was 82+/-13.69x10(-3) mm2/s in the atypical group and 92.21+/-13.21x10(-3) mm2/s in the typical group. No statistically significant differences were found between the 2 groups. Two subtypes of typical meningiomas, secretory and angiomatous meningiomas, had the highest values in the ADC maps. In the histological analysis, there was a significant association between tumor cellularity and the signal in the ADC map. CONCLUSION: Meningiomas show moderately restricted diffusion. The signal on the ADC map is associated with tumor cellularity but we have not demonstrated its usefulness for predicting the histological grade. PMID- 19552930 TI - Haematological and serum biochemical reference values for apparently healthy working horses in Pakistan. AB - This study aimed to establish comprehensive reference values for horses working in developing countries, and to compare them against accepted values for horses in developed countries, supporting diagnosis and clinical decision-making. Horses in developing countries usually perform strenuous work in hot, resource-limited conditions, so their 'normal' blood parameters may differ from other horses. Blood was analysed from 203 working horses in Pakistan, meeting defined clinical criteria. Age, sex, body condition and work-type showed small significant effects, but none were clinically relevant. Of the 32 reference intervals, 28 overlapped those of UK horses. However, the entire reference interval for creatine kinase was higher than for UK horses, while those for erythrocytes, albumin and albumin:globulin ratio were lower. Haematocrit and haemoglobin concentrations were also low. Therefore, apparently healthy working horses may have chronic muscle damage from overwork, and may have sub-clinical anaemia. Interventions combating these conditions could improve animal welfare, although it is unclear whether differences between UK and Pakistan reference values reflect chronic abnormalities, or are in fact physiological adaptations enabling horses to cope with the challenging conditions. PMID- 19552931 TI - Occurrence of canine parvovirus type 2c in the dogs with haemorrhagic enteritis in India. AB - Canine parvovirus 2 (CPV-2) causes a highly contagious and often fatal disease in dogs. Since its sudden emergence in the early 1970s, CPV-2 has been evolving through the generation of novel genetic and antigenic variants (CPV-2a/b/c) that are unevenly distributed throughout the world. In the present study we have examined 36 clinical cases of dogs suspected of CPV collected during year 2006. A fragment of the VP2 gene of the virus was analyzed using polymerase chain reaction (PCR), restriction endonuclease (RE) and DNA sequence analysis. Out of the 36 samples analyzed, 16 were found positive for CPV-2a/2b by conventional PCR. DNA sequencing was done for 6 PCR positive samples, out of which three were characterized as CPV-2c, indicating that this CPV type 2c is currently circulating in India. PMID- 19552932 TI - Nitric oxide production in acute gastroenteritis in Indian children. AB - This study compared nitric oxide (NO) levels in 110 children with, and 110 children without, infectious gastroenteritis. Post-infection intestinal function was assessed in a subset. At least one pathogen was identified in 47.2% of cases. The most common diarrhoeal pathogens were rotavirus (22.7%) and norovirus genogroup II (11.8%). The levels of NO measured by median urinary nitrite:creatinine ratio were significantly higher in children with diarrhoea [23.6; interquartile range (IQR) 12.3-46.7] than without diarrhoea (7.8; IQR 4.1 13.2), P<0.001. The ratio was not significantly different between diarrhoeal cases with and without pathogens (P=0.148). Six of twelve children tested had intestinal dysfunction. PMID- 19552934 TI - Microbiological water quality along the Danube River: integrating data from two whole-river surveys and a transnational monitoring network. AB - The River Danube is, with 2780 km, the second longest river in Europe. Its catchment area covers 801 500 km(2), with approximately 81 million inhabitants in 19 countries. River water for anthropogenic use, transportation and recreation is of major importance in all of these countries. Microbiological contamination from faecal pollution by anthropogenic sources is considered to be a crucial problem throughout the Danube River basin. Thus, detailed knowledge on the extent and the origin of microbial pollution is essential for watershed management. The determination of faecal indicator concentrations along the Danube and its major tributaries during two whole-river surveys and 16 permanent stations allowed for the first time to draw a clear picture of the faecal pollution patterns along the whole longitudinal profile of this important international river. By including a variety of environmental variables in statistical analysis, an integrative picture of faecal pollution in the Danube River basin could be evolved. Four hot spots and six stretches of differing faecal pollution were identified, mainly linked with input from large municipalities. Significant decline of microbiological pollution was observed in the upper and lower Danube stretches over the investigation period. In contrast, a significant increase in the middle part was evident. The planned implementation of new wastewater treatment plants and advanced wastewater treatment measures according to the European Union urban wastewater directive will have a great potential to reduce microbial faecal pollution in the Danube and thus improving water quality. PMID- 19552933 TI - HPV16 E7 oncogene expression in normal human epithelial cells causes molecular changes indicative of an epithelial to mesenchymal transition. AB - Cancer-associated epithelial to mesenchymal transition (EMT) is crucial for invasion and metastasis. Molecular hallmarks of EMT include down-regulation of the epithelial adhesion protein E-cadherin and de-novo expression of N-cadherin and the mesenchymal intermediate filament proteins vimentin and fibronectin. Expression of HPV16 E7 in normal human epithelial cells caused increased levels of vimentin and fibronectin, whereas the epithelial adhesion protein E-cadherin was expressed at decreased levels. Similar expression patterns of vimentin, fibronectin and E-cadherin were also detected in cells expressing HPV16 E6 and E7 or the entire HPV16 early transcriptional unit. HPV16 E6 and E7 were each able to induce N-cadherin expression. Interestingly, these changes in expression levels of EMT-associated proteins are not similarly reflected at the level of mRNA expression, suggesting that HPV16 oncoproteins also modulate EMT through non transcriptional mechanisms. Hence, HPV16 oncoproteins may contribute to malignant progression through EMT induction. PMID- 19552935 TI - Robotic surgical telepathology between the Iron Mountain and Milwaukee Department of Veterans Affairs Medical Centers: a 12-year experience. AB - Since mid-1996, we have operated a diagnostic robotic telepathology (TP) system at the Iron Mountain, MI, Department of Veterans Affairs Medical Center (VAMC) from the Milwaukee, WI, VAMC, located some 220 miles away. No on-site pathologist is present in Iron Mountain. Instead, an experienced, well-trained pathologist assistant, under direction of pathologists located in Milwaukee, is responsible for tissue grossing and sectioning. The pathologist assistant places slides onto the stage of the robotic microscope, which is then controlled by pathologists in Milwaukee. Each case read by TP is subsequently read by light microscopy (LM) by the same pathologist. Three distinct phases of TP have been recognized. Our experience during phase I (mid-1996 to early 1999) has been published previously. During phase II (early 1999 to mid-2004), 1 of the 2 senior telepathologists in phase I retired, and 3 junior pathologists were hired. During phase III (mid-2004 to June 2008), 2 new junior pathologists were hired, and ASAP Imaging (Apollo Telemedicine, Inc., Falls Church, VA) was implemented. The number of TP case opportunities in phases I, II, and III was 2200, 5841, and 3512, respectively, resulting in a total of 11 553. A total of 1834 cases were deferred to LM for a variety of reasons. The number of TP diagnoses rendered in phases I, II, and III was 2144, 4636, and 2939, respectively, resulting in a total of 9719. The major discordance rates in phases I, II, and III were 0.33%, 0.45%, and 0.20%, respectively, with an overall rate of 0.35%. Pathologist-specific discordance rates were not significantly different and ranged from a low of 0.12% to a high of 0.77%, whereas case deferral rates were significantly different (P < .0001) and ranged from 2.5% to 28.7%. In general, no relationship between deferral rate and discordance rate was noted. Iron Mountain clinicians have expressed great satisfaction with the services provided by their off-site pathologist colleagues. PMID- 19552936 TI - Competency assessment of residents in surgical pathology using virtual microscopy. AB - Our goal was to develop an efficient and reliable performance-based virtual slide competency examination in general surgical pathology that objectively measures pathology resident's morphologic diagnostic skill. A Perl scripted MySQL database was used to develop the test editor and test interface. Virtual slides were created with the Aperio ScanScope. The examination consisted of 20 questions using 20 virtual slides. Slides were chosen to represent general surgical pathology specimens from a variety of organ systems. The examination was administered in a secure environment and was completed in 1 to 1 1/2 hours. Examination reliability, as an indicator of the test's ability to discriminate between trainee ability levels, was excellent (r = 0.84). The linear correlation coefficient of virtual slide competency examination score versus months of surgical pathology training was 0.83 (P = .0001). The learning curve was much steeper early in training. Correlation of virtual slide competency examination performance with resident's performance on the 64 item Resident In-Service Examination surgical pathology subsection was 0.70. Correlation of virtual slide competency examination performance with global end of rotation ratings was 0.28. This pilot implementation demonstrates that it is possible to create a short, reliable performance-based assessment tool for measuring morphologic diagnostic skill using a virtual slide competency examination. Furthermore, the examination as implemented in our program will be a valid measure of an individual resident's progress in morphologic competency. Virtual slide technology and computer accessibility have advanced to the point that the virtual slide competency examination model implemented in our program could have applicability across multiple residency programs. PMID- 19552937 TI - Overview of telepathology, virtual microscopy, and whole slide imaging: prospects for the future. AB - Telepathology, the practice of pathology at a long distance, has advanced continuously since 1986. Today, fourth-generation telepathology systems, so called virtual slide telepathology systems, are being used for education applications. Both conventional and innovative surgical pathology diagnostic services are being designed and implemented as well. The technology has been commercialized by more than 30 companies in Asia, the United States, and Europe. Early adopters of telepathology have been laboratories with special challenges in providing anatomic pathology services, ranging from the need to provide anatomic pathology services at great distances to the use of the technology to increase efficiency of services between hospitals less than a mile apart. As to what often happens in medicine, early adopters of new technologies are professionals who create model programs that are successful and then stimulate the creation of infrastructure (ie, reimbursement, telecommunications, information technologies, and so on) that forms the platforms for entry of later, mainstream, adopters. The trend at medical schools, in the United States, is to go entirely digital for their pathology courses, discarding their student light microscopes, and building virtual slide laboratories. This may create a generation of pathology trainees who prefer digital pathology imaging over the traditional hands-on light microscopy. The creation of standards for virtual slide telepathology is early in its development but accelerating. The field of telepathology has now reached a tipping point at which major corporations now investing in the technology will insist that standards be created for pathology digital imaging as a value added business proposition. A key to success in teleradiology, already a growth industry, has been the implementation of standards for digital radiology imaging. Telepathology is already the enabling technology for new, innovative laboratory services. Examples include STAT QA surgical pathology second opinions at a distance and a telehealth-enabled rapid breast care service. The innovative bundling of telemammography, telepathology, and teleoncology services may represent a new paradigm in breast care that helps address the serious issue of fragmentation of breast cancer care in the United States and elsewhere. Legal and regulatory issues in telepathology are being addressed and are regarded as a potential catalyst for the next wave of telepathology advances, applications, and implementations. PMID- 19552938 TI - Virtual slide telepathology enables an innovative telehealth rapid breast care clinic. AB - An innovative telemedicine-enabled rapid breast care service is described that bundles telemammography, telepathology, and teleoncology services into a single day process. The service is called the UltraClinics Process. Because the core services are at 4 different physical locations, a challenge has been to obtain stat second opinion readouts on newly diagnosed breast cancer cases. To provide same day quality assurance rereview of breast surgical pathology cases, a DMetrix DX-40 ultrarapid virtual slide scanner (DMetrix Inc, Tucson, AZ) was installed at the participating laboratory. Glass slides of breast cancer and breast hyperplasia cases were scanned the same day the slides were produced by the University Physicians Healthcare Hospital histology laboratory. Virtual slide telepathology was used for stat quality assurance readouts at University Medical Center, 6 miles away. There was complete concurrence with the primary diagnosis in 139 (90.3%) of cases. There were 4 (2.3%) major discrepancies, which would have resulted in a different therapy and 3 (1.9%) minor discrepancies. Three cases (1.9%) were deferred for immunohistochemistry. In 2 cases (1.3%), the case was deferred for examination of the glass slides by the reviewing pathologists at University Medical Center. We conclude that the virtual slide telepathology quality assurance program found a small number of significant diagnostic discrepancies. The virtual slide telepathology program service increased the job satisfaction of subspecialty pathologists without special training in breast pathology, assigned to cover the general surgical pathology service at a small satellite university hospital. PMID- 19552940 TI - Ovarian carcinoma pathology and genetics: recent advances. AB - In this review we summarize recent advances in the histopathological diagnostic criteria and molecular pathology of the main subtypes of ovarian surface epithelial carcinoma. These advances have greatly improved our understanding of the biology of ovarian carcinoma and are also relevant to patient management. With progress toward subtype-specific treatment of ovarian carcinoma, accurate, reproducible histopathological diagnosis of these subtypes by practicing pathologists is increasingly important. PMID- 19552939 TI - Virtual slide telepathology workstation of the future: lessons learned from teleradiology. AB - The clinical reading environment for the 21st century pathologist looks very different than it did even a few short years ago. Glass slides are quickly being replaced by digital "virtual slides," and the traditional light microscope is being replaced by the computer display. There are numerous questions that arise however when deciding exactly what this new digital display viewing environment will be like. Choosing a workstation for daily use in the interpretation of digital pathology images can be a very daunting task. Radiology went digital nearly 20 years ago and faced many of the same challenges so there are lessons to be learned from these experiences. One major lesson is that there is no "one size fits all" workstation so users must consider a variety of factors when choosing a workstation. In this article, we summarize some of the potentially critical elements in a pathology workstation and the characteristics one should be aware of and look for in the selection of one. Issues pertaining to both hardware and software aspects of medical workstations will be reviewed particularly as they may impact the interpretation process. PMID- 19552941 TI - Ascorbic acid, garlic extract and taurine alleviate cadmium-induced oxidative stress in freshwater catfish (Clarias batrachus). AB - An experiment was conducted to investigate bioaccumulation potential of cadmium (Cd) and changes in oxidative stress indices in liver and kidney tissues from Cd exposed catfish (Clarias batrachus) with or without simultaneous treatment of water with ascorbic acid, garlic extract or taurine. C. batrachus (n=324) with average length of 20+/-4 cm and weight of 86+/-5 g were used for the present investigation. Fishes were divided into nine groups (I to IX) each comprising 36 fishes. The fishes of groups II, III, IV and V were challenged with 5 ppm of cadmium chloride monohydrate (CdCl2.H2O), whereas groups VI, VII, VIII and IX were exposed to 10 ppm CdCl2.H2O solution for a period of 45 days. Group I was kept as negative control and the fishes of this group were maintained in water containing no added Cadmium. Group II and VI were maintained as Cd exposed non treated control to serve as positive controls. Fishes of III and VII, IV and VIII, V and IX received ascorbic acid (5 ppm), extract of dried garlic (5 ppm) or taurine (5 ppm), respectively during the entire experiment period. The concentrations of Cd in liver and kidney increased significantly following exposure to Cd and the level continued to rise with the increase in exposure duration. Treatment of tank water with ascorbic acid, garlic or taurine significantly reduced the Cd concentrations in tissues compared to the positive control group, but the level in Cd exposed groups was greater than the negative control group. Fishes exposed to Cd and treated with ascorbic acid, garlic or taurine had reduced oxidative stress as evidenced from lower concentration of lipid peroxides and higher activities of superoxide dismutase and catalase in liver, kidney and erythrocytes compared to fishes exposed to Cd. The reduction in Cd induced oxidative stress was highest in ascorbic acid treated group followed by garlic and taurine treatment. The results suggest that ascorbic acid, garlic and taurine have potential to reduce tissue accumulation of Cd and associated oxidative stress in freshwater catfish. PMID- 19552942 TI - DGT estimates cadmium accumulation in wheat and potato from phosphate fertilizer applications. AB - Cadmium is a common impurity in phosphatic fertilizers and may contribute to soil Cd accumulation. Changes in total and bioavailable Cd burdens to agricultural soils and the potential for plant Cd accumulation resulting from fertilizer input was investigated. Three year field studies were conducted using three dose levels of cadmium-rich, commercial, phosphate fertilizers applied at four agricultural sites. Labile Cd concentrations, measured using the passive sampling device Diffusive Gradients in Thin Films (Cd(DGT)), increased with increasing fertilizer application rates. Cd also accumulated in the edible portion of wheat and potato crops grown at the sites, and showed strong positive dose response with fertilizer treatment. Regression models were calculated for each site, year, and for individual crops. Model comparisons indicated that soil physical and chemical parameters in addition to soil Cd fractions, were important determinants of Cd(DGT). Significant factors contributing to Cd(DGT) concentrations were Cd from fertilizer input (Cd(fertilizer)), pH, cation exchange capacity (CEC), and total recoverable Cd (Cd(total)). Important factors used to determine Cd concentrations in wheat grain (Cd(wheat)) and in potato (Cd(potato)) were as follows: Cd(wheat):Cd(fertilizer), and Cd(DGT); and Cd(potato):Cd(fertilizer), Cd(DGT), % O.M. The effective concentration, C(E), calculated from DGT did not correlate well with Cd(wheat) or with Cd(potato). Direct measurements of Cd(DGT) correlated better with Cd found in edible plant tissue. The modeling approach presented in this study helps to estimate Cd accumulation in plant tissue over multiple years and in distinct agricultural soil systems. PMID- 19552943 TI - Reproductive toxicity of nitroaromatics to the cricket, Acheta domesticus. AB - The effect of TNT (2,4,6-trinitrotoluene) and its metabolites, 2,4-dinitrotoluene (2,4-DNT), 2-amino-4,6-dinitrotoluene (2A-DNT), and 4-amino-2,6-dinitrotoluene (4A-DNT) on cricket (Acheta domesticus) reproduction was evaluated. We previously used crickets to assess the toxicity of a nitramine explosive (RDX) and its metabolites. It is common to find that while much information on the environmental impact of the parent compound is available in the literature, such is often not the case for the degradation metabolites of the parent compound. In some instances, these metabolites are as toxic (or more so) as the parent compound and we hypothesized that this might be the case for TNT. The presence of TNT and its metabolites in sand (10 microg/g) did not adversely affect cricket egg production, but adversely affected hatching of cricket eggs as compared to controls. However, there were no differences in hatching success among TNT and metabolite treatment groups. Hatching success of cricket eggs in soil or following topical exposure decreased as concentrations of TNT and its metabolites increased. The relative toxicity of TNT and its metabolites in soil generally followed the trend of TNT<2A-DNT<4A-DNT<2,4-DNT. In addition, toxicity appeared to be higher in sand than in sandy loam soil or in the topical exposure test. After 45 days of exposure in sandy loam soil, the EC(20) (20% effect concentration), EC(50) (50% effect concentration), and EC(95) (95% effect concentration) were 14, 116, and 10,837 microg/g for TNT: 1.7, 32, and 16,711 microg/g for 2A-DNT: 1.9, 9, and 296 microg/g for 4A-DNT: and 0.4, 5.7, and 1437 microg/g for 2,4-DNT. Overall, results suggest that parent TNT and metabolites are toxic to cricket eggs at relatively high concentrations and these toxic effects are manifested as a decrease in hatching success. PMID- 19552944 TI - The risk of gastrointestinal perforation and/or fistula in patients with recurrent ovarian cancer receiving bevacizumab compared to standard chemotherapy: a retrospective cohort study. AB - OBJECTIVE: To determine the rate of gastrointestinal perforation and/or fistula in patients with recurrent ovarian cancer treated with and without bevacizumab. METHODS: A retrospective chart review from January 2004 to August 2007 identified two cohorts of patients with recurrent ovarian cancer: 1) patients who were receiving bevacizumab either alone or in combination with standard chemotherapy; 2) patients who were receiving standard chemotherapy alone. Gastrointestinal toxicity (perforation and fistula) was assessed using NCI Common Toxicity Criteria. Relative risk and 95% confidence intervals were calculated. Chi square test and student's t test were used for statistical analysis. RESULTS: Sixty eight patients receiving bevacizumab for recurrent ovarian cancer were identified. 67% of these patients received chemotherapy in combination with bevacizumab. For comparison, 195 patients receiving standard chemotherapy alone for recurrent ovarian cancer were identified. A history of previous gastrointestinal resection (40% vs. 37%; p=0.79) and gastrointestinal obstruction (30% vs. 27%; p=0.74) was similar in both cohorts. Five patients (7.2%) developed a gastrointestinal perforation and/or fistula in the bevacizumab cohort compared to 13 patients (6.5%) in the chemotherapy alone cohort. The relative risk for developing a perforation and/or fistula is 1.09 (95% CI, 0.40 to 2.96). CONCLUSIONS: Although a substantial number of patients with recurrent ovarian cancer experience gastrointestinal obstruction, the rate of gastrointestinal perforation and/or fistula is relatively low. Treatment with bevacizumab does not significantly increase gastrointestinal toxicity compared to standard salvage chemotherapy. PMID- 19552945 TI - Self-reported sexual debut and behavior in young adults aged 18-24 years in seven European countries: implications for HPV vaccination programs. AB - OBJECTIVE: To generate country-specific data on age of sexual debut in young adults in 7 European countries with regard to the optimal age for prophylactic human papillomavirus (HPV) vaccination. METHODS: Survey of self-reported sexual debut and behavior in a sample of young adults aged 18-24 years in 7 European countries (Czech Republic, Ireland, Italy, Netherlands, Poland, Russia, France). Subjects (minimum of 500 males and 500 females per country) were recruited at public places using an in-street collecting approach in all countries except France, where the survey was conducted at home. Data were collected using a short, anonymous, self-administered questionnaire designed to gather information about any kind of sexual activity the subject might have engaged in with a partner, with standardized content to enable comparison between countries. RESULTS: Up to 14% of young men and 22% of young women aged 18-24 years had yet to experience sexual activity at the time of the survey. Median age of sexual debut calculated by survival analysis ranged between 16 and 17 years in boys and between 17 and 18 years in girls. The proportion of boys sexually active before the age of 15 years ranged from 5.0% (Poland) to 15.8% (Italy). The proportion of girls sexually active before the age of 15 years was lower compared with boys in all countries, ranging from 2.6% (Poland) to 11.9% (the Netherlands). The first sexual partner for girls was typically at least 1 year older, irrespective of the age of sexual debut. Almost one-third of young women did not use a condom at sexual debut. CONCLUSIONS: This survey provides an update on sexual debut and behavior in young adults in some European countries. Estimated age at sexual debut ranged between 16 and 18 years and appeared to be later in girls than in boys. The proportion of girls sexually active before the age of 15 years was low (/=48 months) with 93% specificity and 100% sensitivity, and with a positive predictive value of 100% and a negative predictive value of 98%. INTERPRETATION: Stage-adapted management with surgery for stage I disease, and combined surgery and chemotherapy for stage II, III, and IV disease could improve the effectiveness of treatment for placental-site trophoblastic tumours. Use of 48 months since antecedent pregnancy as a prognostic indicator of survival could help select patients for risk-adapted treatment. FUNDING: National Commissioning Group. PMID- 19552950 TI - Moderation of prekallkrein-factor XII interactions in surface activation of coagulation by protein-adsorption competition. AB - Traditional biochemistry of contact activation of blood coagulation suggesting that anionic hydrophilic surfaces are specific activators of the cascade is inconsistent with known trends in protein adsorption. To investigate contact activation reactions, a chromogenic assay was used to measure prekallkrein (PK) hydrolysis to kallikrein (Kal) by activated factor XII (FXIIa) at test hydrophilic (clean glass) and hydrophobic (silanized glass) surfaces in the presence of bovine serum albumin (BSA). Hydrolysis of PK by FXIIa is detected after contact of the zymogen FXII with a test hydrophobic surface only if putatively-adsorbed FXIIa is competitively displaced by BSA. By contrast, FXIIa activity is detected spontaneously following FXII activation by a hydrophilic surface and requires no adsorption displacement. These results (i) show that an anionic hydrophilic surface is not a necessary cofactor for FXIIa-mediated hydrolysis of PK, (ii) indicate that PK hydrolysis does not need to occur by an activation complex assembled directly on an anionic, activating surface, (iii) confirms that contact activation of FXII (autoactivation) is not specific to anionic hydrophilic surfaces, and (iv) demonstrates that protein-adsorption competition is an essential feature that must be included in any comprehensive mechanism of surface-induced blood coagulation. PMID- 19552951 TI - An acid-labile temperature-responsive sol-gel reversible polymer for enhanced gene delivery to the myocardium and skeletal muscle cells. AB - The work demonstrates the development of acid-labile temperature-responsive sol gel reversible polymer for enhanced in vivo myocardium and skeletal muscle gene delivery. In this report, multi-block copolymers (MBCPs) synthesized from pluronic and di-(ethylene glycol) divinyl ether (DEGDVE) were used as a delivery vehicle for controlled and sustained release of plasmid DNA (pDNA) in in vitro as well as in vivo experiments. The non-ionic MBCP/pDNA complex showed remarkable transfection efficiencies against the myocardium cells as well as muscle cells in vivo, which is otherwise very difficult to achieve by using cationic polymers. In in vitro experimental settings, this intelligent stimuli-responsive polymer is shown to improve the transfection efficiency of branched polyethylenimine (BPEI)/pDNA complex when used together. The effect of MBCP on the surface charge and particle size of its various complexes with pDNA and BPEI was also studied. The release profile of pDNA from the MBCP gel was investigated and pH of the degraded polymer was also monitored to ascertain its non-cytotoxicity arising due to the increased acidity as observed with other PLGA-based polymers. The rapid sol-gel transition of MBCP under thermal stimuli with concomitant release of pDNA under acidic stimulation has potential for site specific, efficient and controlled transfection of therapeutic gene. In short, MBCP provides the silver lining in combat against the hurdles encountered in transfection to myocardial or other muscle cells. PMID- 19552952 TI - Calcium alginate beads embedded in silk fibroin as 3D dual drug releasing scaffolds. AB - 3D scaffolds based on embedding drug loaded calcium alginate beads within silk fibroin protein were fabricated for investigating controlled dual drug release. The 3D matrices were evaluated for in vitro release using two different molecular weight model compounds, bovine serum albumin (66 kDa) and FITC-Inulin (3.9 kDa). The model compound release profiles revealed dependence on molecular weight of encapsulated model drugs for sustained release. Further, silk fibroin protein blended calcium alginate beads resulted in prolonged drug release without initial bursts for 35 days as compared to calcium alginate beads without silk fibroin as control. The release kinetics were further tested as a function of wt% silk content for scaffold fabrication suggesting their possible role in restricting initial burst and leading to sustainable release of compounds for prolong time. Silk coatings on calcium alginate beads provided mechanically stable shells as well as a diffusion barrier to the encapsulated protein drugs thus controlling their release. Scanning electron microscopic observations were carried out to assess cellular viability and biocompatibility of bead embedded-silk 3D scaffolds using fibroblast cells. The results highlight the versatile and tunable properties of calcium alginate embedded fibroin 3D scaffolds making them exciting candidate for the controlled release of a wide spectrum of bioactive molecules from a single delivery vehicle. PMID- 19552953 TI - The role of nitric oxide in the inhibitory effect of ghrelin against penicillin induced epileptiform activity in rat. AB - Ghrelin, a gastric peptide with key action on food intake, has been recently recognized as a potential antiepileptic agent. In the present study, we investigated the involvement of nitric oxide in the effect of ghrelin on penicillin-induced epileptiform activity in rat. Thirty minutes after penicillin injection, ghrelin, at doses of 0.5, 1, 2 microg, was administered intracerebroventricularly (i.c.v.). Ghrelin, at a dose of 1 microg, significantly decreased the mean frequency of epileptiform activity without changing the amplitude whereas other doses of ghrelin (0.5 and 2 microg) did not alter either the mean of frequency or amplitude of epileptiform activity. The effects of systemic administration of nitric oxide synthase (NOS) inhibitors, non-selective N(G)-nitro-l-arginine methyl ester (l-NAME), selective neuronal NOS inhibitor, 7 nitroindazole (7-NI) and NO substrate, l-arginine on the anticonvulsive effects of ghrelin were investigated. The administration of l-NAME (60 mg/kg, i.p.), 15 min before ghrelin (1microg) application, reversed the anti-epileptiform effects of ghrelin whereas 7-NI (40 mg/kg, i.p.) did not influence it. The present study provides electrophysiological evidence that the intracerebroventricular injection of ghrelin has an inhibitory effect against epileptiform activity in the penicillin model of epilepsy. The anti-epileptiform activity of ghrelin was reversed by nonspecific nitric oxide synthase inhibitor l-NAME, but not selective neuronal nitric oxide synthase inhibitor 7-NI, indicating that ghrelin requires activation of endothelial-NOS/NO route in the brain. PMID- 19552954 TI - Pediatric primary care as a component of systems of care. AB - Systems of care should be defined in a manner that includes primary care. The current definition of systems of care shares several attributes with the definition of primary care: both are defined as community-based services that are accessible, accountable, comprehensive, coordinated, culturally competent, and family focused. However, systems of care is defined as serving only children and youth with serious emotional disturbance and their families and does not fully embrace the concept of primary prevention. Although similarities in the definitions of primary care and systems of care may provide a theoretical foundation for including primary care within the systems of care framework, a definition of systems of care that incorporates the idea of prevention and takes into account the broad population served in primary care would provide communities with a definition that can be used to further the work of integrating primary care into systems of care. PMID- 19552955 TI - Evolutionary drift in systems of care development. AB - While we agree that it is necessary to articulate a "clearly specified population" for the definition of systems of care, we believe that limiting systems of care to "children and youth with serious emotional disturbance and their families" is not in the best interest of most communities. Using this narrow population definition excludes the other 80-90% of the youth who have mental health challenges but have not risen to the highest level of need. If all children and their families receive the right amount of support from the system of care at the right time, they will avoid the need for more intensive and expensive services and supports later. This public health approach helps to build more stable communities and redirects scarce resources to interventions that are less costly than those needed for youth who already have developed serious emotional disturbances. The key to successfully supplying the "right amount at the right time" is to ensure that the system of care is truly needs driven, rather than agency or service system driven. A system of care for children, youth and their families should reflect community preferences and embrace a public health approach where all levels of need are served. PMID- 19552956 TI - Getting a grip on systems of care and child welfare using opposable thumbs. AB - The purpose of this response paper is to discuss issues raised by two of the components of the definition of systems of care proffered by Hodges et al. [Hodges, S., Ferreira, K., Israel, N., & Mazza, J. (this issue). Systems of care, featherless bipeds, and the measure of all things. Evaluation and Program Planning]. In particular, this response will present some implications of the definition of the focus population and the value and core principle of family driven care. It will also consider why these two components of the definition might serve as challenges to the applicability of the concept of systems of care to child welfare, and, in turn, integration of the model across child welfare and mental health. Recommendations for expanding and refining these component terms are provided. PMID- 19552957 TI - From complexity to reality: providing useful frameworks for defining systems of care. AB - Because systems of care are not uniform across communities, there is a need to better document the process of system development, define the complexity, and describe the development of the structures, processes, and relationships within communities engaged in system transformation. By doing so, we begin to identify the necessary and sufficient components that, at minimum, move us from usual care within a naturally occurring system to a true system of care. Further, by documenting and measuring the degree to which key components are operating, we may be able to identify the most successful strategies in creating system reform. The theory of change and logic model offer a useful framework for communities to begin the adaptive work necessary to effect true transformation. Using the experience of two system of care communities, this new definition and the utility of a theory of change and logic model framework for defining local system transformation efforts will be discussed. Implications for the field, including the need to further examine the natural progression of systems change and to create quantifiable measures of transformation, will be raised as new challenges for the evolving system of care movement. PMID- 19552958 TI - Challenges to parenting in a new culture: Implications for child and family welfare. AB - Increasing numbers of families arriving through Australia's humanitarian settlement scheme are coming into contact with Australian child protection systems. A large number of these families come from African and Middle Eastern countries and have common experiences of trauma, dislocation, loss and many are victims of genocide, war, and torture. Pre-migration experiences together with the considerable challenges of settling into a new country can significantly affect family well-being and parenting practices. It is therefore important that child and family welfare service planners are well informed about how best to support refugee families using culturally competent family intervention and community development practices. This paper draws on the findings of a research project designed to examine why recently arrived families from refugee backgrounds are presenting in the South Australian child protection system and to identify culturally appropriate strategies for intervention. The paper presents findings from the project that relate to (1) refugee parents', community members' and child protection practitioners' perspectives on the challenges to being a refugee parent in Australia and (2) strategies and resources relevant to prevention and early intervention in refugee families before statutory child protection intervention becomes necessary. PMID- 19552959 TI - Defining "necessary" services and supports: why systems of care must take direction from service-level processes. AB - A crucial element of the system of care definition is the specification of its purpose, namely, "to ensure access to and availability of necessary services and supports." This article discusses the structures and processes that must be in place so that systems of care can acquire and respond to high-quality information about what services and supports are truly necessary. PMID- 19552961 TI - A global study of pathogens and host risk factors associated with infectious gastrointestinal disease in returned international travellers. AB - OBJECTIVES: Infectious gastrointestinal disease (IGD) is a significant cause of morbidity in returned international travellers. This study aims to elucidate host and travel characteristics associated with IGD presentation, and describe the broad spectrum of aetiological pathogens responsible by geographic region of acquisition and reason for travel. METHODS: We analyzed demographic, clinical and microbiological data recorded for ill, returned international travellers presenting to GeoSentinel Surveillance Network sites globally during the period September 1996-December 2005. RESULTS: A total of 25,867 returned travellers were analyzed, of whom 7442 (29%) patients had a total of 8273 IGD diagnoses. Multivariate analysis demonstrated that IGD presentation was associated significantly with female sex (OR: 1.11; p=0.001); younger age group; attending a pre-travel medical appointment (OR: 1.28; p<0.0001); and travelling for the reason of tourism. Travelling for longer periods (>28 days) was associated with lower risk (OR: 0.93; p=0.04). Of the 2902 clinically significant pathogens isolated, 65% were parasitic, 31% bacterial and 3% viral. Presentation of IGD by specific pathogen varied markedly dependent on geographic region of recent travel, and reason for travel. CONCLUSIONS: Host characteristics, region of travel and category of traveller, significantly impact on the relative likelihood of presenting with a broad range of pathogen-specific IGD. PMID- 19552962 TI - The modified SCID Hypomania Module (SCID-Hba): a detailed systematic phenomenologic probing. AB - Diagnosing past hypomania is a difficult task. Current structured interviews (e.g. CIDI, SCID) limit the ability to probe for hypomania. A modified SCID Hypomania Module was published by us (Benazzi and Akiskal, J Affect Disord 2003; Akiskal and Benazzi, J Clin Psychiatry 2005) in order to overcome the limitations of structured interviewing. Our papers outlined the framework of the modified SCID. In response to requests from many readers of this journal and other clinicians and investigators, we are hereby providing a more explicit step-by step phenomenologic probing interview. DSM-IV criteria have to be met, but the probing for hypomania is very different from that of the SCID. All past hypomanic symptoms are assessed. No negative meaning is given to symptoms, as hypomania often improves functioning and it is seen by patients as a state of well being. The first step is probing for overactivity (increase in goal-directed activity), because observable behaviors are easier to remember by patients and key informants. There is no gold-standard for overactivity: each person becomes his/her own standard to 'measure' a clear-cut departure form the usual behavior. Questions, correspondingly, can change from patient to patient. The emotions associated with behavioral change are easier to be remembered than asking them first, as in the structured interviews. Structured interviews have mood change (elation, irritability) as stem question (corresponding to the criterion A of DSM IV, which postulates that it must always be present). However, apart from a likely recall bias of past emotions, the description of mood change appears more or less negative in structured interviews (to increase specificity but by much reducing sensitivity, i.e. the false-negatives). Presenting mood change as simply having been more elated/irritable than usual can easily be interpreted as normal mood fluctuations, while presenting mood change as much more than usual could be understood as a severe mental disorder. Both ways are likely to lead to a negative response, moving the interviewers to unipolar disorders (the skip-out instruction). Our modified SCID is a fully semi-structured interview: many questions are asked about each symptom to make the question understandable according to each patient, and, very importantly, examples of the 'events' are systematically asked to check understanding and clinical relevance. Our interview follows DSM-IV criteria (apart from the minimum duration, 2 days versus DSM-IV 4 days), i.e. mood change must always been present, but our probing detects more hypomanic episodes than the SCID. PMID- 19552960 TI - Protective KIR-HLA interactions for HCV infection in intravenous drug users. AB - Intravenous drug use has become the principal route of hepatitis C virus (HCV) transmission due to the sharing of infected needles. In this study, we analyzed the distribution of HLA-KIR genotypes among 160 Puerto Rican intravenous drug users (IDUs) with HCV infection and 92 HCV-negative Puerto Rican IDUs. We found a significant association between the presence of different combinations of KIR inhibitory receptor genes (KIR2DL2 and/or KIR2DL3, pC=0.01, OR=0.07; KIR2DL2 and/or KIR2DL3+KIR2DS4, pC=0.01, OR=0.39) and HLA-C1 homozygous genotypes (HLA C1+KIR2DS4, pC=0.02, OR=0.43; HLA-C1+KIR2DL2+KIR2DS4, pC=0.02, OR=0.40) together with the activating receptor KIR2DS4 (HLA-C1+KIR2DS4+KIR2DL3 and/or KIR2DL2, pC=0.004, OR=0.38) with protection from HCV infection. Our findings in HCV infected and non-infected IDUs suggest an important role for KIRs (KIR2DL2 and KIR2DL3) with group HLA-C1 molecules, in the presence of activating KIR2DS4, in protection from HCV infection. These results support the hypothesis that activator signaling, mediated by KIR2DS4, plays a determinant role in the regulation of NK cell antiviral-activity. PMID- 19552963 TI - Efficacy and safety of paliperidone extended-release in schizophrenia patients with prominent affective symptoms. AB - BACKGROUND: This post-hoc analysis evaluated the effects of paliperidone extended release (ER) in patients with schizophrenia and prominent affective symptoms. METHODS: Pooled data from three 6-week, randomized, double-blind, placebo controlled studies were analyzed. Subjects received fixed doses of paliperidone ER 3-12 mg/day or placebo. Prominent affective symptoms were defined as depressive (Positive and Negative Syndrome Scale [PANSS] depression item score of > or =5 [moderately severe]) and/or manic (PANSS grandiosity score of > or =4 [moderate], plus a score of > or =4 [moderate] on at least 1 PANSS item for excitement, hostility, uncooperativeness, or poor impulse control). Assessments included PANSS, Clinical Global Impressions-Severity (CGI-S), Personal and Social Performance (PSP) scale, and adverse events (AEs). RESULTS: Among 193 patients with prominent affective symptoms, 140 received paliperidone ER and 53 received placebo. Paliperidone ER showed significant mean (SD) improvements vs. placebo in PANSS total (-20.5 [23.8] vs. -6.3 [27.2]; p<0.001, respectively) and all factor scores (p<0.01). Significant mean (SD) improvements were observed in PSP (7.2 [15.8] vs. 0.4 [14.6]; p=0.004) and CGI-S (-0.9 [1.2] vs. -0.3 [1.2]; p<0.001) scores. Most common AEs with paliperidone ER vs. placebo: headache (16.4% vs. 13.2%), insomnia (7.9% vs. 9.4%), akathisia (7.1% vs. 1.9%), sedation (7.1% vs. 3.8%). LIMITATIONS: These studies were not designed to examine patients with prominent affective symptoms. Authors' clinical judgment was used to define prominent affective symptoms, using relevant PANSS items. CONCLUSIONS: Paliperidone ER was well tolerated and associated with significantly greater improvements in symptomatology, functioning, and overall clinical status vs. placebo in patients with schizophrenia and prominent affective symptoms. PMID- 19552965 TI - Isolation of lysozyme and an antifungal peptide from sea lamprey (Petromyzon marinus) plasma. AB - The sea lamprey (Petromyzon marinus) belongs to the most primitive class of fish and has only innate immunity. The innate immune factors, lysozyme and an antifungal peptide, were isolated from sea lamprey plasma. Sea lamprey plasma (40.1mg protein/ml) was assayed for lysozyme activity by gel diffusion assay. Using hen egg white lysozyme standards, plasma concentration of lamprey lysozyme was 5microg lysozyme/mg total protein. The presence of lysozyme in such high concentration in lamprey plasma could be important in their innate immunity and resistance to infection. Lysozyme and the antifungal peptide were isolated by low molecular weight gel filtration chromatography from sea lamprey plasma. Gel filtration chromatography yielded two peak tubes containing lysozyme (1microg/211microg total protein) and antifungal peptide (1microg/66microg total protein). Lysozyme and antifungal activity of each fraction were determined by well diffusion assay using Gram-negative bacteria, Gram-positive bacteria and two fungal species. The molecular weight of lamprey lysozyme was 14.3kDa. The sea lamprey lysozyme was effective against Gram-positive bacteria but not against Gram-negative bacteria or fungi. Molecular weight of the antifungal peptide was approximately 3000Da. Antifungal plasma activity was seen against Penicillium notatum and Aspergillus flavus. No plasma antibacterial peptide was found. PMID- 19552966 TI - Diverging effects of cortistatin and somatostatin on the production and release of prostanoids from rat cortical microglia and astrocytes. AB - Here we compared the effects of cortistatin and somatostatin on the production of prostanoids from primary cultures of rat cortical microglia and astrocytes. We found that both cortistatin and somatostatin do not modify basal PGE(2) release from cultured astrocytes in 24-h experiments. Somatostatin further enhanced the increase in PGE(2) release induced by IL-1beta, whereas cortistatin inhibited such increase. Experiments on microglia showed that somatostatin has no effect on basal and IL-1beta-stimulated PGE(2) release, whereas cortistatin reduced baseline prostanoids production and abolished stimulation elicited by IL-1beta. The latter effect was associated to the inhibition of COX-2 gene over-expression induced by the cytokine. PMID- 19552967 TI - History and development of pediatric otorhinolaryngology in Hungary. AB - The first pediatric otorhinolaryngological department was set up in Hungary at the Heim Pal Children's Hospital, Budapest in 1948. The first head of the department and the founder of Hungarian pediatric otorhinolaryngology was Ferenc Kallay. He was followed in leadership by Jeno Hirschberg, and at present Gabor Katona. In Budapest 10 pediatric otolaryngological departments and in other parts of the country seven departments have been working until recently with overall 344 beds. The Pediatric Otorhinolaryngological Section was developed in 1977. The discipline is independent specialty since 1978. Conferences with various special topics have been organized every year since then. Two international congresses were held in Hungary: the first one in Eger, 1986 presided by Jeno Hirschberg and Zoltan Labas; and the second in Budapest in 2008, organized by Gabor Katona. The Hungarian pediatric otorhinolaryngologists assumed initiative role in the development of this special discipline six decades ago, early joined in the international life and have had important positions in several international associations (IFOS, ESPO, UEP, IALP, IAP) as president, board member or chairman of committees. Besides the organizational work, they have taken part in the scientific and research work as well: in the present paper the authors detail the results achieved by them in several topics of pediatric otorhinolaryngology. PMID- 19552968 TI - Recombinant antibody therapeutics: the impact of glycosylation on mechanisms of action. AB - More than twenty recombinant antibody molecules are now licensed for the treatment of a variety of cancers and chronic diseases. Initially, the attraction of antibodies was their specificity for target antigens; however, it is now appreciated that the downstream consequences of engaging antigen, after the formation of immune complexes, is crucial to clinical outcomes in vivo. This review introduces the structural and functional activities of the IgG class of recombinant antibodies, in vitro, and criteria that determine choice between the four subclasses. Importantly, we demonstrate that, although accounting for only 2 3% of antibody mass, glycosylation of the IgG-Fc is essential to the activation of downstream biologic mechanisms (effector functions). Additionally the precise structure of the attached oligosaccharide can influence biologic efficacy. These findings have led to cellular engineering to enable the production of selected glycoforms of antibody that are considered to be optimal for the disease indication to be treated. PMID- 19552969 TI - The relationship of force output characteristics during a sit-to-stand movement with lower limb muscle mass and knee joint extension in the elderly. AB - This study examined the reliability of ground reaction force (GRF) parameters during sit-to-stand (STS) movements and the relationships between the GRF parameters and lower limb muscle mass and knee extension muscle strength. Fifty elderly females performed an STS movement twice from a chair adjusted to their knee height and the GRF, lower limb muscle mass and isometric knee extension muscle strength were measured. Reliabilities of GRF parameters were high (intra class correlation coefficient=ICC=0.70-0.95). Parameters on force output during trunk flexion phase (ground reaction force at hip-lift off, sum of force output between beginning of STS movement and hip-lift off) differed significantly between trials, but their effect sizes were small (0.15-0.23). GRF parameters during hip-lift off and knee-hip joint extension phases significantly correlated with knee extension strength (|r|=0.29-0.64) but not lower limb muscle mass. In conclusion, the reliability of GRF during STS movement is good in hip-lift off and knee-hip joint extension phases and these phases relate significantly with lower limb muscle function. These two phases are useful for evaluation of leg muscle function of the elderly. PMID- 19552970 TI - Interest and limits of glomerular filtration rate (GFR) estimation with formulae using creatinine or cystatin C in the malnourished elderly population. AB - Renal function is often altered in elderly patients. A lot of formulae are proposed to estimate GFR to adjust drug posology. French guidelines recommend the Cockcroft-Gault formula corrected with the body surface area (cCG), but the initially described unadjusted Cockcroft-Gault equation (CG) is mainly used in geriatric clinical practice. International recommendations have proposed the modification of diet in renal disease (MDRD) formula, since several authors recommended the Rule formula using cystatin C (cystC) in particular population. To appreciate the most accurate GFR estimation for posology adaptation in an elderly polypathological population, a cross-sectional study with prospective inclusion was carried out in Charles Foix Hospital. Plasma glucose levels (PGL), creatinine (CREA) levels and serum cystC, albumin (ALB), transthyretin (TTR), C reactive protein (CRP), orosomucoid (ORO) total cholesterol (tCHOL) levels were determined among 193 elderly patients aged 70 and older. The results showed that in a malnourished, inflamed old population, CG, MDRD and Rule formulae resulted in different estimations of GFR, depending on nutritional and inflammatory parameters. Only cCG estimation was shown to be independent from these parameters. To conclude, cCG seems to be the most accurate and appropriate formula in a polypathological elderly population to evaluate renal function in order to adapt drug posology. PMID- 19552971 TI - How to assess high erythrocyte sedimentation rate (ESR) in elderly? AB - The ESR is commonly used to assess the acute phase response. It is most useful among patients in whom the probability of disease is moderate following initial history-taking and examination. In this study, we examined retrospectively the patients admitted to our hospital to lighten the cause of a high ESR and accompanying anemia and compared the results between the geriatric population and younger adults. There were 139 patients between ages of 16 and 89 years. Of them, 51.7% were elderly. In 80 patients (57.6%) a specific underlying pathology as a possible cause of elevated ESR was found. Malignancy was the leading cause (21.6%), followed by infectious disorders (10.1%), collagen vascular diseases (9.4%), and non-neoplastic hematologic disorders (5.0%). In 59 patients (42.4%) no specific pathology could be found. There were no statistically significant differences between elderly and non-elderly patients according to the diagnostic groups. Elevated ESRs, while more prevalent in the elderly than in younger individuals, have a similar pathological significance. But as clinical conditions in the elderly are usually obscure, we should be more careful to investigate the issue of high ESR in geriatric population, even when they are asymptomatic. PMID- 19552972 TI - Comparison and age-level differences among various step tests for evaluating balance ability in the elderly. AB - This study aimed to examine the difficulty among various step tests (place step, forward single step, forward double step, forward right single step and stairs step) in evaluating the dynamic balance in the elderly and their age level differences. Thirty-two healthy elderly people (age 71.4+/-6.4 years) and twenty young people performed step tests for 10 s to the pace of a metronome (120 bpm). Evaluation parameters were the time difference between the metronome sound and the time when each foot hit the ground as well as the stride time. The forward single step test had significantly larger values for both of the above parameters than the other tests. A significant age level difference was found in the forward single step test for the time difference and in the forward single step and stairs step tests for the stride time, being longer in the elderly. It was concluded that the forward single step test has larger age-level differences and is more difficult to carry out than the other step tests. PMID- 19552973 TI - Acute worsening in migraine symptoms following PFO closure: a matter of fact? AB - There is currently conjecture in the literature as to whether percutaneous closure of patent foramen ovales (PFO) leads to an improvement in migraine symptoms. The present study reports the migraine status at 30 days, of 57 consecutive patients who underwent closure PFO closures for cryptogenic stroke at our institution. Our findings suggest a significant change in migraine character following PFO closure with the majority of these patients reporting a worsening in migraine severity. Those patients with more severe migraine prior to the procedure were more likely to experience deterioration. Patients should be counselled about the possibility of changing or worsening migraine prior to percutaneous closure for cryptogenic stroke. PMID- 19552974 TI - Fund for non-fast food. PMID- 19552976 TI - Usefulness of a 0.010-inch guidewire compatible balloon for crossing stent-jailed sidebranches. PMID- 19552977 TI - Space-time Bayesian survival modeling of chronic wasting disease in deer. AB - The primary objectives of this study are to describe the spatial and temporal variation in disease prevalence of chronic wasting disease (CWD), to assess the effect of demographic factors such as age and sex on disease prevalence and to model the disease clustering effects over space and time. We propose a Bayesian hierarchical survival model where latent parameters capture temporal and spatial trends in disease incidence, incorporating several individual covariates and random effects. The model is applied to a data set which consists of 65085 harvested deer in Wisconsin from 2002 to 2006. We found significant sex effects, spatial effects, temporal effects and spatio-temporal interacted effects in CWD infection in deer in Wisconsin. The risk of infection for male deer was significantly higher than that of female deer, and CWD has been significantly different over space, time, and space and time based on the harvest samples. PMID- 19552975 TI - L-carnitine reduces doxorubicin-induced apoptosis through a prostacyclin-mediated pathway in neonatal rat cardiomyocytes. AB - BACKGROUND: Clinical use of doxorubicin is greatly limited by its severe cardiotoxic side effects. L-carnitine is a vitamin-like substance which has been successfully used in many cardiomyopathies, however, the intracellular mechanism(s) remain unclear. The objective of this study was set to evaluate the protective effect of L-carnitine on doxorubicin-induced cardiomyocyte apoptosis, and to explore its intracellular mechanism(s). METHODS: Primary cultured neonatal rat cardiomyocytes were treated with doxorubicin (1 uM) with or without pretreatment with L-carnitine (1-30 mM). Lactate dehydrogenase assay, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling staining, and flow cytometry measurement were used to assess cytotoxicity and apoptosis. Fluorescent probes 2',7'-dichlorofluorescein diacetate and chemiluminescence assay of superoxide production were used to detect the production of reactive oxygen species. Western blotting was used to evaluate the quantity of cleaved caspase-3, cytosol cytochrome c, and Bcl-x(L) expression. RESULTS: L-carnitine inhibited doxorubicin-induced reactive oxygen species generation and NADPH oxidase activation, reduced the quantity of cleaved caspase 3 and cytosol cytochrome c, and increased Bcl-x(L) expression, resulting in protecting cardiomyocytes from doxorubicin-induced apoptosis. In addition, L carnitine was found to increase the prostacyclin (PGI(2)) generation in cardiomyocytes. The siRNA transfection for PGI(2) synthase significantly reduced L-carnitine-induced PGI(2) and L-carnitine's protective effect. Furthermore, blockade the potential PGI(2) receptors, including PGI(2) receptors (IP receptors), and peroxisome proliferator-activated receptors alpha and delta (PPARalpha and PPARdelta), revealed that the siRNA-mediated blockage of PPARalpha considerably reduced the anti-apoptotic effect of L-carnitine. CONCLUSIONS: These findings suggest that L-carnitine protects cardiomyocytes from doxorubicin induced apoptosis in part through PGI(2) and PPARalpha-signaling pathways, which may potentially protect the heart from the severe toxicity of doxorubicin. PMID- 19552978 TI - Functional image-based radiotherapy planning for non-small cell lung cancer: A simulation study. AB - BACKGROUND AND PURPOSE: To investigate the incorporation of data from single photon emission computed tomography (SPECT) or hyperpolarized helium-3 magnetic resonance imaging ((3)He-MRI) into intensity-modulated radiotherapy (IMRT) planning for non-small cell lung cancer (NSCLC). MATERIAL AND METHODS: Seven scenarios were simulated that represent cases of NSCLC with significant functional lung defects. Two independent IMRT plans were produced for each scenario; one to minimise total lung volume receiving >or=20Gy (V(20)), and the other to minimise only the functional lung volume receiving >or=20Gy (FV(20)). Dose-volume characteristics and a plan quality index related to planning target volume coverage by the 95% isodose (V(PTV95)/FV(20)) were compared between anatomical and functional plans using the Wilcoxon signed ranks test. RESULTS: Compared to anatomical IMRT plans, functional planning reduced FV(20) (median 2.7%, range 0.6-3.5%, p=0.02), and total lung V(20) (median 1.5%, 0.5-2.7%, p=0.02), with a small reduction in mean functional lung dose (median 0.4Gy, 0 0.7Gy, p=0.03). There were no significant differences in target volume coverage or organ-at-risk doses. Plan quality index was improved for functional plans (median increase 1.4, range 0-11.8, p=0.02). CONCLUSIONS: Statistically significant reductions in FV(20), V(20) and mean functional lung dose are possible when IMRT planning is supplemented by functional information derived from SPECT or (3)He-MRI. PMID- 19552979 TI - Rapid delivery of stereotactic radiotherapy for peripheral lung tumors using volumetric intensity-modulated arcs. AB - The delivery of high dose conventional stereotactic body radiotherapy (SBRT) for patients with stage I lung tumors generally takes 30-45min per fraction. The novel volumetric intensity-modulated arc therapy (RA) for planning and delivery enabled much faster treatment for three patients with different fractionation schemes. This reduces the risk of intrafraction motion and is more patient friendly. In addition, in comparison to the conventional plans using 10 static non-coplanar fields, RA plans achieved superior dose conformity around the PTV and reduced chest wall doses. PMID- 19552980 TI - Fiducial-free real-time image-guided robotic radiosurgery for tumors of the sacrum/pelvis. AB - BACKGROUND AND PURPOSE: Radiosurgery is a non-invasive treatment for many spinal tumors. Sacral radiosurgery, however, requires invasive fiducial marker insertion to target and track the tumor's position. We present preliminary clinical results and phantom accuracy measurements of sacral radiosurgery using fiducial-free alignment based on vertebral anatomy distant to tumor location. MATERIALS AND METHODS: Fifty-one lesions in 38 patients were treated using fiducial-free spinal tracking of the L5 vertebra. An anthropomorphic phantom was used for accuracy measurements of this approach. Dose planning was based on 1.0mm computer tomography slices using inverse treatment planning. RESULTS: Tracked targets were up to 17cm from the treated tumor. Phantom tests produced an overall mean targeting error of 1.43mm (+/-0.47mm). Patient median follow-up was 12.7months. Local tumor control was 95%. Treatment doses were 12-25Gy with a median prescription isodose of 65% (40-70%) and tumor volumes between 1.3 and 152.8cc. No short-term adverse events were noted during the follow-up period. CONCLUSIONS: Fiducial-free tracking of the lower lumbar vertebrae is a feasible, accurate, and reliable tool for radiosurgery of sacral and pelvic tumors. It is a valuable novel alternative to surgical procedures and conventional fractionated radiation therapy for these challenging cases. PMID- 19552981 TI - The effect of calcium on the transcriptome of sporulating B. subtilis cells. AB - Bacterial spores formed in the presence of high concentrations of minerals are a major problem in the food industry because of their extreme heat resistance. In order to enhance our insight in the molecular mechanisms underlying this phenomenon we have performed a detailed time-resolved analysis of the genome-wide transcriptome pattern of Bacillus subtilis sporulated both in the absence and presence of high calcium concentrations. The data was analysed in two ways. First, we determined the influence of the presence of high calcium levels during sporulation on the expression of gene groups as defined in Subtilist and KEGG pathways database. Second, we assessed the differential expression at the level of individual genes. When analysing groups and pathways, we found that those annotated as being involved in sporulation were significantly affected. Also, groups and pathways involved in flagella formation and biofilm matrix production were affected by the presence of calcium in the sporulation medium. When we analysed the behaviour of individual genes we found 305 genes influenced by calcium, including all known spore coat polysaccharide biosynthesis genes (10 induced and 1 repressed). A number of the calcium affected genes were also involved in biofilm formation. Minimal overlap with other stress outputs like sigma B activation and weak acid stress response was noted. Those genes that did overlap were unique to that combination which corroborates the notion that the cells sense these conditions differently. PMID- 19552982 TI - Hospital investment policy in France: pathways to efficiency and the efficiency of the pathways. AB - OBJECTIVES: This article examines the ambivalent notion of New Public Management as applied to health policy in France, by investigating the implementation of the efficiency-driven hospital investment plan, Hopital 2012, conceived at national level, but implemented through regional hospital authorities (ARHs), with formal responsibility for selecting successful funding applications. METHODS: The methodology combines qualitative and quantitative analysis, in order to highlight and explain discrepancies between goals and results. RESULTS: Despite formal adherence to objective efficiency indicators, certain decisions were based on incomplete information and others on considerations out with initially established criteria. Competition from the private sector was perceived as a threat to public hospitals and the public sector emerged as a major beneficiary of the investment plan. Central ministerial intervention emphasising financial and quantitative considerations led the ARHs to focus more on individual hospital performance than on wider healthcare needs. CONCLUSIONS: Data-production became almost an end in itself, threatening to undermine the objectives it sought to pursue. Nonetheless, extended deadlines entailed by ministerial intervention were appropriated as a resource by local actors, leading to ARH decisions which deviated from the official efficiency model, but resulted in increased effectiveness, taking fuller account of local conditions. PMID- 19552984 TI - Inhibitory effects of 5-benzylidene barbiturate derivatives on mushroom tyrosinase and their antibacterial activities. AB - A series of novel 5-benzylidene barbiturate and thiobarbiturate derivatives were synthesized and evaluated as tyrosinase inhibitors and antibacterial agents. The results demonstrated that some compounds had more potent inhibitory activities than the parent compound 4-hydroxybenzaldehyde (IC(50)=1.22 mM). Particularly, compounds 1a and 2a were found to be the most potent inhibitors with IC(50) value of 13.98 microM and 14.49 microM, respectively. The inhibition mechanism study revealed that these compounds were irreversible inhibitors. The circular dichroism spectra indicated that these compounds induced conformational changes of mushroom tyrosinase upon binding. In addition, these compounds exhibited selectively antibacterial activity against Staphylococcus aureus. All these results suggested that further development of such compounds may be of interest. PMID- 19552985 TI - New insights on pneumococcal disease: what we have learned over the past decade. AB - Since the introduction of the seven-valent pneumococcal conjugate vaccine (PCV7) in 2000, significant reductions in the rate of invasive pneumococcal diseases (IPDs) and mucosal infections caused by vaccine-specific serotypes have been observed in the targeted population (children aged <5 years). IPD rates have also decreased in adults aged >/=65 years, suggesting the extension of vaccine protection to the unimmunized population. A concomitant decline has been noted in antibiotic-resistant Streptococcus pneumoniae isolates. The incidence of vaccine serotype IPD varies widely in geographically disparate regions, indicating that serotype coverage alone may not accurately predict the impact of proposed pneumococcal conjugate vaccines with additional serotypes. PMID- 19552986 TI - Mass balance approach to estimating radionuclide loads and concentrations in edible fish tissues using stable analogues. AB - Humans can consume a number of types of biota tissues, which have varying propensities to accumulate radionuclides. As a result, depending upon the biota species, the radionuclide and the tissue under consideration, it may be necessary to estimate the percent radionuclide load in specific edible tissues, and in cases where whole organisms are consumed, to estimate the radionuclide load in the whole body of an organism, based on data that have been collected for individual tissues. To accomplish this, data were compiled that can be used to estimate the partitioning patterns and percent loads of various groups of elements in edible tissues of freshwater fishes. General trends in partitioning, such as those provided in this paper, can be used to predict radionuclide transfer to humans and the corresponding potential radiological dose to humans via dietary pathways, in this case following the consumption of fish. PMID- 19552987 TI - Laboratory and field studies of polonium and plutonium in marine plankton. AB - Uptake experiments were successfully undertaken for polonium and plutonium in the marine diatom Skeletonema costatum. Experiments were complemented with a field study which yielded concentration factors for these radionuclides for plankton samples collected in the eastern Irish Sea. The uptake experiment produced uptake half times of 0.8 and 0.6 days for plutonium and polonium, respectively. Concentration factors of 3.1 x 10(5) l kg(-1) for plutonium and 2.4 x 10(5) l kg( 1) for polonium were consistent with those suggested by the IAEA for phytoplankton. Concentration factors derived from the field study were 2 x 10(3) l kg(-1) for plutonium and 1.4 x 10(4) l kg(-1) for polonium and were more consistent with IAEA values for zooplankton rather than phytoplankton. The maximum calculated dose rate due to Po and Pu in plankton sampled in the field was 0.12 microGyh(-1). In applying the laboratory-derived phytoplankton CF values for Po and Pu, higher dose rates, in the range 6-11 microGyh(-1), were calculated. The latter values are well below the UNSCEAR and IAEA review value of 400 microGyh(-1), and are also below the Environment Agency marine action level of 40 microGyh(-1). PMID- 19552988 TI - An evaluation of the Africa Midwives Research Network. AB - OBJECTIVE: to evaluate the strengths and limitations of the Africa Midwives Research Network (AMRN) and provide feedback and direction to the network and the funding body. DESIGN: a qualitative study incorporating visits to three African countries and Sweden, using interviews, non-participant observation, an internet survey and review of records. SETTING: principally Tanzania, Uganda, Zambia and Sweden. FINDINGS: AMRN can be described as a small, dispersed, loosely coupled professional network which has made considerable impact on midwifery practice in the areas of its members. The biennial scientific conferences could be perceived as AMRN's flagship activity and have been notably successful, becoming renowned internationally. Around 1500 midwives have benefited from educational programmes at regional or national level. These include research methodology, evidence-based practice, scientific writing and communication skills. Attention needs to be given to some aspects of governance and organization, as well as to communication strategies including websites and newsletters. Technical support from the Karolinska Institute in Stockholm has provided good collegiate support and proved to be cost effective. KEY CONCLUSIONS: AMRN has shown resilience and continuity since its inception and has made a palpable difference to the quality of midwifery care and the professional development of midwives within the remit of its members. AMRN needs to be consolidated before expanding further. The work of AMRN is particularly pertinent in the context of the millennium development goals. PMID- 19552989 TI - Long-term air monitoring of organochlorine pesticides using Semi Permeable Membrane Devices (SPMDs) in the Alps. AB - Atmospheric sampling of organochlorine pesticides (OCPs) was conducted using Semi Permeable Membrane Devices (SPMDs) deployed in the Alps at different altitudinal transects for two consecutive exposure periods of half a year and a third simultaneous year-long period. Along all the altitude profiles, the sequestered amounts of OCPs increased in general with altitude. SPMDs were still working as kinetic samplers after half a year for the majority of the OCPs. However, compounds with the lowest octanol-air partition coefficient (K(oa)), reached equilibrium within six months. This change in the SPMD uptake was determined for the temperature gradient along the altitude profile influencing K(oa), OCPs availability in the gaseous phase, and SPMD performance. In sum, it seems two effects are working in parallel along the altitude profiles: the change in SPMD performance and the different availability of OCPs along the altitudinal transects determined by their compound properties and concentrations in air. PMID- 19552990 TI - Cognitive vulnerability to anxiety: A review and an integrative model. AB - Consistent research evidence supports the existence of threat-relevant cognitive bias in anxiety, but there remains controversy about which stages of information processing are most important in the conferral of cognitive vulnerability to anxiety. To account for both theoretical and empirical discrepancies in the literature, an integrative multi-process model is proposed wherein core assumptions of dual-systems theories from social and cognitive psychology are adapted to explain attentional and interpretive biases in the anxiety disorders. According to the model, individual differences in associative and rule-based processing jointly influence orientation, engagement, disengagement, and avoidance of threat-relevant stimuli, as well as negatively-biased interpretation of ambiguous stimuli in anxious populations. By linking anxiety-related symptoms to basic principles of information processing, the model parsimoniously integrates different kinds of cognitive biases in anxiety, providing a useful framework for future research and clinical intervention. PMID- 19552991 TI - From prenatal HIV testing of the mother to prevention of sexual HIV transmission within the couple. AB - The first step in preventing mother-to-child HIV transmission (PMTCT) programmes is offering HIV counselling and testing to pregnant women. In developing countries where HIV testing remains rare, it represents a unique opportunity for many women to learn their HIV status. This prenatal HIV testing is not only the entry point to prevention of mother-to-child HIV transmission, but also an occasion for women to sensitize their male partner to sexual risks. Here we explore if these women, HIV-tested as mothers, apply the prevention recommendations they also receive as women. In the Ditrame Plus PMTCT program in Abidjan, Cote d'Ivoire, two cohorts of women (475 HIV-infected women and 400 HIV negative women) were followed up two years after the pregnancy when they were offered prenatal HIV testing. In each cohort, we compared the proportion of women who communicated with their regular partner on sexual risks, prior to and after prenatal HIV testing. We analysed socio-demographic factors related to this communication. We measured two potential conjugal outcomes of women HIV testing: the level of condom use at sex resumption after delivery and the risk of union break-up. Prenatal HIV testing increased conjugal communication regarding sexual risks, whatever the woman's serostatus. This communication was less frequent for women in a polygamous union or not residing with their partner. Around 30% of women systematically used condoms at sex resumption. Among HIV infected ones, conjugal talk on sexual risks was related to improved condom use. After HIV testing, more HIV-infected women separated from their partners than HIV uninfected women, despite very few negative reactions from the notified partners. In conclusion, offering prenatal HIV counselling and testing is an efficient tool for sensitizing women and their partners to HIV prevention. But sexual prevention in a conjugal context remains difficult and need to be specifically addressed. PMID- 19552992 TI - Relative deprivation in income and self-rated health in the United States. AB - Absolute income is robustly associated with health status. Few studies have, however, examined if relative income is independently associated with health. We examined if, over and above the effects of absolute income, individual relative deprivation in income as well as position in the income hierarchy is associated with individual poor health in the U.S. Using three rounds of the Current Population Surveys (CPS), we analyzed the association between self-rated health (1=fair/poor, 0=otherwise) and the Yitzhaki index of relative deprivation in income and percentile position in the income hierarchy across 17 reference groups. Over and above the effects of absolute income, the odds ratio for reporting poor health among individuals in the highest quintile of relative deprivation compared to the lowest quintile ranged between 2.18 and 3.30, depending on the reference groups used. A 10 percentile increase in income position within reference groups was associated with an odds ratio of poor health of 0.89. Relative deprivation appeared to explain between 33 and 94% of the association between individual income and self-rated health. Relative deprivation in income is independently associated with poor health over and above the well established effects of absolute income on health. Relative deprivation may partly explain the association between income inequality and worse population health status. PMID- 19552993 TI - Occupational career and risk of mortality among US Civil War veterans. AB - Previous studies have extended the traditional framework on occupational disparities in health by examining mortality differentials from a career perspective. Few studies, however, have examined the relation between career and mortality in a historical U.S. population. This study explores the relation between occupational career and risk of mortality in old age among 7096 Union Army veterans who fought the American Civil War in the 1860s. Occupational mobility was commonplace among the veterans in the postbellum period, with 54% of them changing occupations from the time of enlistment to 1900. Among veterans who were farmers at enlistment, 46% of them changed to a non-farming occupation by the time of 1900. Results from the Cox Proportional Hazard analysis suggest that relative to the average mortality risk of the sample, being a farmer at enlistment or circa 1900 are both associated with a lower risk of mortality in old age, although the effect is more salient for veterans who were farmers at enlistment. Occupational immobility for manual labors poses a serious threat to chance of survival in old age. These findings still hold after adjusting for the effects of selected variables characterizing risk exposures during early life, wartime, and old age. The robustness of the survival advantage associated with being a farmer at enlistment highlights the importance of socioeconomic conditions early in life in chance of survival at older ages. PMID- 19552994 TI - [Long term effect of autoadjusting positive airway pressure on C-reactive protein and interleukin-6 in men with obstructive sleep apnoea syndrome]. AB - BACKGROUND AND OBJECTIVES: Obstructive sleep apnoea (OSA) has been increasingly linked to cardiovascular disease. Inflammatory processes associated with OSA may contribute to this morbidity. Some studies have reported serum levels of high sensitivity C-reactive protein (hs-CRP) and interleukin-6 (IL-6) to be increased in these patients. PRIMARY OBJECTIVE: investigate the impact of short and long term autoadjusting positive airway pressure (APAP) therapy on IL-6 and hs-CRP serum levels in patients with moderate to severe obstructive sleep apnoea. Secundary Objective: evaluate the basal hs-CRP and IL-6 levels in OSA patients and its possible relation to OSA severity, independently of confounders and compare the hs-CRP levels in OSA patients with those in community controls. PATIENTS AND METHODS: This is a prospective study including 98 male patients with moderate to severe OSA confirmed by domiciliary sleep study. Malignancy and chronic inflammatory diseases were exclusion criteria. hs-CRP and IL-6 serum levels were evaluated before APAP, 9 days and 6 months after therapy. Community controls (n=103) were selected using random digit dialing, and matched by age and body mass index (BMI) for comparison of hs-CRP levels at baseline. RESULTS: The studied population had a mean age of 55.3+/-10.7 years, mean BMI 33.2+/ 5.0kg/m(2), mean apnoea-hypopnoea index 51.7+/-21.3/h and mean desaturation index 86.3+/-5.3/h. The APAP compliance was good: 91.27%+/-20.45 days usage and 5.76+/ 1.59h/night of usage. Mean basal hs-CRP and IL-6 serum values were 0.52+/ 0.53microg/l and 17.7+/-22.5microg/l, respectively. CRP levels at baseline correlated significantly with apnoea-hypopnoea index, desaturation index and minimum nocturnal oxygen saturation. IL-6 levels at baseline correlated significantly and negatively with minimum nocturnal oxygen saturation. When adjusting for confounding factors found in this study, all these relations lost significance. CRP is significantly increased in patients when compared to controls (p=0.002) and when considering hs-CRP cardiovascular risk stratified categories, cases had significantly more patients at high risk of cardiovascular events than controls (p=0.002). After adjustment for BMI and arterial hypertension, cases had an almost twofold moderate risk of cardiovascular events and more than a twofold severe risk of cardiovascular events when compared to controls. We found no significant difference between hs-CRP and IL-6 concentrations pre-treatment and in two moments post-treatment (9 days and 6 months) (CRP: p=0.720 and p=0.387, respectively; IL-6: p=0.266 and p=0.238, respectively). CONCLUSIONS: OSA is associated with a low-grade inflammatory process; hs-CRP serum levels are elevated in OSA patients when comparing to community controls, independently of age and BMI and the former have a significantly higher risk of cardiovascular events when compared to the latter. There was no significant decrease of both inflammatory mediators (hs-CRP, IL-6) after short and long-term APAP therapy. PMID- 19552995 TI - [Pneumonia due to Dialister pneumosintes in a patient with chronic obstructive pulmonary disease]. PMID- 19552997 TI - Detecting evidence of luteal activity by least-squares quantitative basal temperature analysis against urinary progesterone metabolites and the effect of wake-time variability. AB - OBJECTIVE: To assess computerised least-squares analysis of quantitative basal temperature (LS-BT) against urinary pregnanediol glucuronide (PdG) as an indirect measure of ovulation, and to evaluate the stability of LS-QBT to wake-time variation. STUDY DESIGN: Cross-sectional study of 40 healthy, normal-weight, regularly menstruating women aged 19-34. Participants recorded basal temperature and collected first void urine daily for one complete menstrual cycle. Evidence of luteal activity (ELA), an indirect ovulation indicator, was assessed using Kassam's PdG algorithm, which identifies a sustained 3-day PdG rise, and the LS QBT algorithm, by determining whether the temperature curve is significantly biphasic. Cycles were classified as ELA(+) or ELA(-). We explored the need to pre screen for wake-time variations by repeating the analysis using: (A) all recorded temperatures, (B) wake-time adjusted temperatures, (C) temperatures within 2h of average wake-time, and (D) expert reviewed temperatures. RESULTS: Relative to PdG, classification of cycles as ELA(+) was 35 of 36 for LS-QBT methods A and B, 33 of 34 (method C) and 30 of 31 (method D). Classification of cycles as ELA(-) was 1 of 4 (methods A and B) and 0 of 3 (methods C and D). Positive predictive value was 92% for methods A-C and 91% for method D. Negative predictive value was 50% for methods A and B and 0% for methods C and D. Overall accuracy was 90% for methods A and B, 89% for method C and 88% for method D. The day of a significant temperature increase by LS-QBT and the first day of a sustained PdG rise were correlated (r=0.803, 0.741, 0.651, 0.747 for methods A-D, respectively, all p<0.001). CONCLUSION: LS-QBT showed excellent detection of ELA(+) cycles (sensitivity, positive predictive value) but poor detection of ELA(-) cycles (specificity, negative predictive value) relative to urinary PdG. Correlations between the methods and overall accuracy were good and similar for all analyses. Findings suggest that LS-QBT is robust to wake-time variability and that expert interpretation is unnecessary. This method shows promise for use as an epidemiological tool to document cyclic progesterone increase. Further validation relative to daily transvaginal ultrasound is required. PMID- 19552996 TI - Cell death and proliferation in acute slices and organotypic cultures of mammalian CNS. AB - Analysis of the interplay between cell proliferation and death has been greatly advantaged by the development of CNS slice preparations. In slices, interactions between neurons and neurons and the glial cells are fundamentally preserved in a fashion close to the in vivo situation. In parallel, these preparations offer the possibility of an easy experimental manipulation. Two main types of slices are currently in use: the acute slices, which are short living preparations where the major functions of the intact brain (including neurogenesis) are maintained, and the organotypic cultures, where the maturation and plasticity of neuronal circuitries in relation to naturally occurring neuronal death and/or experimental insults can be followed over several weeks in vitro. We will discuss here the main advantages/disadvantages linked to the use of CNS slices for histological analysis of neuronal proliferation and death, as well as the main findings obtained in the most popular types of preparations, i.e. the cortical, hippocampal, cerebellar and retinal slices. PMID- 19552998 TI - Long-term symptomatic group B streptococcal vulvovaginitis: eight cases resolved with freshly cut garlic. PMID- 19552999 TI - A new technique of cervical repair after a laparoscopic enucleation. PMID- 19553000 TI - DTA-mediated targeted ablation revealed differential interdependence of endocrine cell lineages in early development of zebrafish pancreas. AB - Cell lineage analysis is critical in understanding the relationship between progenitors and differentiated cells as well as the mechanism underlying the process of differentiation. In order to study the zebrafish endocrine pancreas cell lineage, transgenic expression of diphtheria toxin gene A chain (DTA) under two cell type-specific promoters derived from the insulin (ins) and somatostatin2 (sst2) genes was used to ablate the two types of endocrine cells: insulin producing beta-cells and somatostatin-producing delta-cells, respectively. We found that ablation of beta-cells resulted in a reduction of not only beta-cells but also glucagon-producing alpha-cells; in contrast, delta-cells were largely unaffected. Ablation of delta-cells led to reduction of all three types of endocrine cells: alpha-, beta-, and delta. Interestingly, alpha-cells were more profoundly affected in both beta- and delta-cell ablations and were frequently reduced together with beta- and delta-cells. By taking advantage of Tg(ins:gfp) and Tg(sst2:gfp) lines, we also monitored the changes of different types of endocrine cells in vivo after ablation and found that both beta- and delta-cell populations significantly recovered by 3dpf after their ablation and it seemed that delta-cells had a better capability of recovery than beta-cells. Thus, our current observations indicated differential interdependence of these three cell lineages. The development of zebrafish alpha-cells, but not delta-cells, is dependent on beta-cells, while the development of both alpha- and beta-cells is dependent on delta-cells. In contrast, the development of delta-cells was independent of beta-cells. PMID- 19553001 TI - Sustainable value assessment of farms using frontier efficiency benchmarks. AB - Appropriate assessment of firm sustainability facilitates actor-driven processes towards sustainable development. The methodology in this paper builds further on two proven methodologies for the assessment of sustainability performance: it combines the sustainable value approach with frontier efficiency benchmarks. The sustainable value methodology tries to relate firm performance to the use of different resources. This approach assesses contributions to corporate sustainability by comparing firm resource productivity with the resource productivity of a benchmark, and this for all resources considered. The efficiency is calculated by estimating the production frontier indicating the maximum feasible production possibilities. In this research, the sustainable value approach is combined with efficiency analysis methods to benchmark sustainability assessment. In this way, the production theoretical underpinnings of efficiency analysis enrich the sustainable value approach. The methodology is presented using two different functional forms: the Cobb-Douglas and the translog functional forms. The simplicity of the Cobb-Douglas functional form as benchmark is very attractive but it lacks flexibility. The translog functional form is more flexible but has the disadvantage that it requires a lot of data to avoid estimation problems. Using frontier methods for deriving firm specific benchmarks has the advantage that the particular situation of each company is taken into account when assessing sustainability. Finally, we showed that the methodology can be used as an integrative sustainability assessment tool for policy measures. PMID- 19553002 TI - Volatilization characteristics of organic solutes in stirred solution. AB - The effects of turbulence intensity (velocity gradient, G (s(-1))), Henry's law constant (H), and molecular weight (M) on the volatilization rates of organic compounds are examined using changes in the mass transfer coefficients (K(OL) (cm/min)) under specific liquid-mixing intensities. The selected compounds were divided into three groups according to their H values (mole in gas/mole in liquid, dimensionless), which ranged from 10(2) to 10(-5). The relationship of the K(OL) relative to G, H and M was obtained via multiple regression. The obtained values of these parameters indicate that the primary factor affecting the K(OL) values of the high H compounds is their M values. The effects of the H values on the K(OL) values of the high H compounds can be neglected. On the other hand, the H value is the major factor determining the K(OL) values of the low H compounds. The changes in the K(OL) values of the different H compounds exhibit different profiles as the liquid-mixing intensity increases. The M and H values of middle H compounds possibly affect their K(OL) values. The effects of the liquid-mixing intensity on the K(OL) values of the organic compounds increase with increasing H values. The variation in the K(OL) values might be a result of the concentration of the organic compounds at the interface between the liquid and gas films. The empirical relationship between K(OL) and some selected parameters, G, H and M, is examined in this study. The obtained results can help to estimate volatilization loss of organic solutes in wastewater treatment facilities. PMID- 19553003 TI - In vivo growth inhibition of head and neck squamous cell carcinoma by the Interferon-inducible gene IFI16. AB - The Interferon-inducible gene, IFI16 has been implicated in the control of cell growth, apoptosis, angiogenesis and immunomodulation. In a previous study we demonstrated that restoring levels of IFI16 in a head and neck squamous cell carcinoma (HNSCC)-derived cell line, HNO136, reduced its growth in vitro accompanied by a marked increase in doxorubicin-induced apoptosis. To evaluate the ability of IFI16 to inhibit in vivo tumorigenesis of HNO136 cells and to characterize the molecular mechanisms responsible for its anti-tumor activity, IFI16 expression on cell growth was evaluated by an in vivo tumorigenicity assay. After excision, tumors were subjected to morphometric and immunohistochemical analyses with markers of apoptosis, angiogenesis, and inflammation. Restoring IFI16 expression significantly reduced the in vivo tumorigenesis of HNO136, decreased tumor vascularization and increased areas of tumor necrosis. Further analysis revealed that IFI16 expression triggered apoptosis of tumor cells, as evaluated using TUNEL assay. Finally, restoring IFI16 protein to HNO136 cells increased CD45+ inflammatory cell infiltration of the tumor burden, predominantly consisting of CD68/CD14 positive macrophages. In accordance with our previous in vitro experiments, this study demonstrates for the first time that IFI16 exerts in vivo anti-tumoral activity by promoting apoptosis of tumor cells, by inhibiting neo-vascularisation, and by increasing the recruitment of macrophages through the release of chemotactic factors. PMID- 19553004 TI - PCC and COBRA-FISH a new tool to characterize primary cervical carcinomas: to assess hall-marks and stage specificity. AB - A newly developed assay based on chemically induced premature chromosome condensation (PCC) and multi-color combined binary ratio labeling (COBRA) fluorescence in situ hybridization (FISH) techniques have been implemented in order to investigate for the first time for recurrent cytogenetic aberrations in primary cervical carcinoma (derived directly from biopsies) at different stages of progression. The cytogenetic profiles of 17 biopsies derived from 14 and 3 cervical cancer patients with squamous-cell carcinomas (Sq) and with adenocarcinomas (Ad), respectively, were assessed. Frequencies of both structural as well as numerical aberrations were found to be higher in Sq than in Ad. The analysis revealed that even in early tumors stages (IB1) have a higher frequency of chromosome-losses and -gains as well as chromosomal alterations as compared to normal cells. A positive trend was found between stage advancement of cervical tumors and the frequency of numerical and structural aberrations. No specific and common chromosomal abnormality (e.g. distinct clones of translocation) was found among cervical carcinoma at the different stages (IB1, IIA and IIB). However, a distinct difference was found between stage IIIB and lower tumor stages, as all analyzed IIIB samples revealed a near tetraploid karyotype. Furthermore, all studied metaphases were aberrant and had a high frequency of translocations. PCC COBRA-FISH characterization of a common type of an established culture from cervical carcinoma CSCC-1 revealed a triploidy/tetraploidy karyotype with several structural aberrations. In general, no similarity was found between this model and early stages of primary tumors. The newly established assay has a novel potential and can reveal the original status of primary tumors at different stages. PMID- 19553005 TI - MKP1 repression is required for the chemosensitizing effects of NF-kappaB and PI3K inhibitors to cisplatin in non-small cell lung cancer. AB - Treatment of non-small cell lung cancer (NSCLC) with cisplatin has a level of antitumor activity still modest. We have shown previously that MKP1/DUSP1 inhibits cisplatin-induced apoptosis in NSCLC cells and is overexpressed in tumors from most patients with stage I-II NSCLC. Here, using different NSCLC cell lines we found that MKP1 and NF-kappaB are differentially expressed. We studied whether targeting MKP1, NF-kappaB or both affects cisplatin-induced cell death. MKP1 is expressed in H460 and H727 cells. H727 and H1299 cells showed constitutive phosphorylation of Akt and increased NF-kappaB activity than did H460 cells. H460-MKP1-siRNA-expressing cells (but not H727-MKP1-siRNA or H1299 MKP1-siRNA cells) exhibit a marked increase in cisplatin response compared with parental cells. Treatment with the PI3K inhibitor LY294002 or the NF-kappaB inhibitor BAY11-7082 enhanced cisplatin antitumor activity in parental H1299 cells but only weakly affected responses of H727 and H460 cells. MKP1-siRNA expression enhanced the chemosensitization effect of LY294002 and BAY11-7082 on H727 and H460 cells. Additionally, NSCLC cell lines with higher NF-kappaB constitutive activation were the most sensitive to PS-341 (Bortezomib), a non specific NF-kappaB inhibitor. This finding suggests the proteasome as a suitable strategy in treating NSCLC tumors with high constitutive NF-kappaB activity. Altogether, these results showed that either an activated PI3K/Akt/NF-kappaB pathway and/or high MKP1 was linked to reduced sensitivity to cisplatin in NSCLC cells. Inhibition of NF-kappaB or PI3K potently enhanced cisplatin cytotoxicity in cells with endogenous or genetically induced low MKP1 levels. These findings support the potential improvement in cisplatin responses by co-targeting NF kappaB or Akt and MKP1. PMID- 19553006 TI - Kinetics and thermodynamic of adsorption of chromium(VI) from aqueous solution using puresorbe. AB - This paper deals with an investigation on coir-based adsorbent, puresorbe, in the removal of chromium(VI) from the aqueous solutions. The adsorption of chromium(VI) was carried out by varying the parameters such as agitation time, metal concentration, adsorbent dose, temperature and pH. The experimental isotherm data were analyzed using Langmuir, Freundlich and Redlich and Peterson isotherms. Adsorption followed second order rate expression for the particle size 250-500 microm at pH 2. The monolayer adsorption capacity is 76.92 mg chromium(VI) per gram of puresorbe. Thermodynamic parameters show the endothermic nature of chromium(VI) adsorption. Desorption study carried out using distilled water adjusted to pH of 2-10, suggests that chemisorption might be the mode of adsorption. PMID- 19553007 TI - Characterizing the effects of shaking intensity on the kinetics of metallic iron dissolution in EDTA. AB - Despite two decades of intensive laboratory investigations, several aspects of contaminant removal from aqueous solutions by elemental iron materials (e.g., in Fe(0)/H2O systems) are not really understood. One of the main reasons for this is the lack of a unified procedure for conducting batch removal experiments. This study gives a qualitative and semi-quantitative characterization of the effect of the mixing intensity on the oxidative dissolution of iron from two Fe(0) materials (materials A and B) in a diluted aqueous ethylenediaminetetraacetic solution (2 mM EDTA). Material A (fillings) was a scrap iron and material B (spherical) a commercial material. The Fe(0)/H2O/EDTA systems were shaken on a rotational shaker at shaking intensities between 0 and 250 min(-1) and the time dependence evolution of the iron concentration was recorded. The systems were characterized by the initial iron dissolution rate (k(EDTA)). The results showed an increased rate of iron dissolution with increasing shaking intensity for both materials. The increased corrosion through shaking was also evidenced through the characterization of the effects of pre-shaking time on k(EDTA) from material A. Altogether, the results disprove the popular assumption that mixing batch experiments is a tool to limit or eliminate diffusion as dominant transport process of contaminant to the Fe(0) surface. PMID- 19553008 TI - Hexavalent chromium [Cr(VI)] removal by acid modified waste activated carbons. AB - Fresh activated carbon (AC) and waste activated carbon (WAC) were pretreated by heating with mineral acids (sulfuric acid and nitric acid) at high temperature to prepare several grades of adsorbents to evaluate their performance on Cr(VI) removal from aqueous phase. Effects of temperature, agitation speed and pH were tested, and optimum conditions were evaluated. Kinetic study was performed under optimum conditions with several grades of modified adsorbents to know the rates of adsorption. Batch adsorption equilibrium data followed both, Freuindlich and Langmuir isotherms. Maximum adsorption capacity (q(max)) of the selected adsorbents treated with sulfuric acid (MWAC 1) and nitric acid (MWAC 2), calculated from Langmuir isotherm are 7.485 and 10.929 mg/g, respectively. Nitric acid treated adsorbent (MWAC 2) was used for column study to determine the constants of bed depth service time (BDST) model for adsorption column design. PMID- 19553009 TI - Sorption behavior of nano-TiO2 for the removal of selenium ions from aqueous solution. AB - Titanium dioxide nanoparticles were employed for the sorption of selenium ions from aqueous solution. The process was studied in detail by varying the sorption time, pH, and temperature. The sorption was found to be fast, and to reach equilibrium basically within 5.0 min. The sorption has been optimized with respect to the pH, maximum sorption has been achieved from solution of pH 2-6. Sorbed Se(IV) and Se(VI) were desorbed with 2.0 mL 0.1 mol L(-1) NaOH. The kinetics and thermodynamics of the sorption of Se(IV) onto nano-TiO2 have been studied. The kinetic experimental data properly correlate with the second-order kinetic model (k(2)=0.69 g mg(-1) min(-1), 293 K). The overall rate process appears to be influenced by both boundary layer diffusion and intraparticle diffusion. The sorption data could be well interpreted by the Langmuir sorption isotherm. The mean energy of adsorption (14.46 kJ mol(-1)) was calculated from the Dubinin-Radushkevich (D-R) adsorption isotherm at room temperature. The thermodynamic parameters for the sorption were also determined, and the DeltaH(0) and DeltaG(0) values indicate exothermic behavior. PMID- 19553010 TI - Comment on "Coupled acidification and ultrasound with iron enhances nitrate reduction" by Tsai et al. [J. Hazard. Mater. 163 (2009) 743]. PMID- 19553011 TI - Extraction of nickel from spent catalyst using fresh and recovered EDTA. AB - This study investigates the possibility of recovering nickel from spent catalyst (NiO/Al(2)O(3)) used in the fertilizer industry. EDTA (ethylenediaminetetraaceticacid) di sodium salt was used as a chelating to extract the Ni after which sulfuric acid was added to obtain NiSO(4). The dechelation process takes 5-6h to break the complex and EDTA which was recovered as H(4)EDTA acid in the solid form and solution contains NiSO(4). The objective was to evaluate the nickel removal efficiency of EDTA and reusability of recovered EDTA. The parameters affecting nickel recovery were EDTA concentration, time of chelation, catalyst to liquid ratio (s:l), mixing speed, pH and catalyst particle size. The extraction was up to 95% under the following conditions: 0.8M concentration of EDTA, solid to liquid ratio 1:50 (g/ml), particle size 100 microm, pH 10, 10h of chelation time, 700 rpm and 100 degrees C. Up to 95% of the EDTA could be recovered without losing significant activity. PMID- 19553012 TI - Effects of dissolved gases and pH on sonolysis of 2,4-dichlorophenol. AB - Ultrasonic degradation of 2,4-dichlorophenol (2,4-DCP) has been studied under oxygen, air, argon, and nitrogen in aqueous solutions for a liquid temperature of 20 degrees C at 489 kHz. The degradation rate increases significantly in the presence of O2, and argon, whereas it remains the lowest under nitrogen, for which a competitive reaction model has been proposed for its non-exponential decay. Experiments have been also performed at three different pHs, 11.0, 6.3, and 2.0. The significant degradation has been achieved at pH 6.3 under O2 (0.86 x 10(-3) s(-1)), which is 1.9 and 4 times higher than acidic (pH 2.0), and basic (pH 11.0) conditions, respectively. The degradation rates have decreased in the order O2 > Ar > air > N2 irrespective of pH. PMID- 19553013 TI - Effect of ultrasound on removal of persistent organic pollutants (POPs) from different types of soils. AB - A new and promising technology is utilization of sonochemistry on decontamination of polluted soil. The feasibility of this technology on treatment of contaminated soils (synthetic clay, natural farm clay, and kaolin) was studied by using two target persistent organic pollutants (POPs): hexachlorobenzene (HCB) and phenanthrene (PHE). The soils were highly contaminated in 500 mg/kg. The laboratory experiments were conducted with various conditions (moisture, power, and time duration). The effects of these parameters on ultrasonication (as well as the removal of contaminants) were examined. The reasonable moisture ratio of the slurry could be in range of 2:1-3:1. The process did not change pH values of soils. Experimental results showed that ultrasonication has a potential to reduce the high concentrations of these POPs. PMID- 19553014 TI - Competitive adsorption of Cu (II), Co (II) and Ni (II) from their binary and tertiary aqueous solutions using chitosan-coated perlite beads as biosorbent. AB - A new composite chitosan-coated biosorbent was prepared and was used for the removal and recovery of heavy metals from aqueous solution. In the present investigation, equilibrium adsorption characteristics of Cu (II), Ni (II), and Co (II) from their binary and tertiary solution on newly developed biosorbent chitosan-coated perlite beads were evaluated through batch and column studies. These beads were characterized by using FTIR, EDXRF and surface area analysis techniques. The effect of various biosorption parameters like effect of pH, agitation time, concentration of adsorbate and amount of adsorbent on extent of adsorption was investigated. The adsorption follows Lagergren first order kinetic model. The equilibrium adsorption data were fitted to Freundlich and Langmuir adsorption isotherm models and the model parameters were evaluated. Both the models represent the experimental data satisfactorily. The sorbent loaded with metal was regenerated with 0.1N NaOH solution. Furthermore the column dynamic studies indicate the re-usage of the biosorbent. PMID- 19553015 TI - A comparative study for the removal of hexavalent chromium from aqueous solution by agriculture wastes' carbons. AB - This paper reports the feasibility of using agricultural waste and timber industry waste carbons to remove Cr(VI) from synthetic wastewater under different experimental conditions. For this, rice husk and saw dust have been used as adsorbent after sulphuric acid treatment. Effect of various process parameters, namely, pH, adsorbent dose, initial chromium concentration and contact time has been studied in batch systems. Maximum metal removal was observed at pH 2.0. The efficiencies of rice husk carbon (RHC) and saw dust carbon (SDC) for Cr(VI) removal were 91.75% and 94.33%, respectively for aqueous solutions (250 mg L(-1)) at 20 g L(-1) adsorbent dose. The experimental data was analyzed using Freundlich, Langmuir, Dubinin-Redushkevich (D-R) and Temkin isotherm models. It was found that Langmuir, D-R and Temkin models fitted well. The results revealed that the hexavalent chromium is considerably adsorbed on RHC and SDC and it could be an economical method for the removal of hexavalent chromium from aqueous systems. FTIR and SEM of the adsorbents were recorded in native and Cr(VI)-loaded state to explore the number and position of various functional groups available for Cr(VI) binding onto studied adsorbents and changes in adsorbent surface morphology. The surface area of RHC and SDC was 1.12 and 1.16 m(2)g(-1), respectively. PMID- 19553017 TI - Comment on: excessive opioid use and the development of chronic migraine. PMID- 19553016 TI - Signaling pathway involved in hypoxia-inducible factor-1alpha regulation in hypoxic-ischemic cortical neurons in vitro. AB - Hypoxia-inducible factor-1alpha (HIF-1alpha) is a key transciptional regulator of cellular and systemic oxygen homeostasis. Previous studies have shown that the regulation of HIF-1alpha is involved in the activation of PI3K/Akt pathway in some cells. However, whether this pathway plays a role in modulating HIF-1alpha in cultured cortical neurons during hypoxia-ischemia (HI) remains unclear. We therefore investigated the relationship between phosphoinositid 3-kinase/Akt (PI3K/Akt) pathway and HIF-1alpha expression in cultured neurons using an oxygen and glucose deprivation (OGD) model. In this study, cortical neurons cultured in vitro were subjected to OGD for 3h followed by reperfusion. The expressions of HIF-1alpha, VEGF, total Akt and phosphorelated-Akt (p-Akt) were detected by RT PCR, Western blot and immunocytochemistry. We found that HIF-1alpha and VEGF were increased at 4h and peaked at 8h after OGD. Meanwhile, p-Akt increased and peaked at 4h after reperfusion, preceding HIF-1alpha expression. Pretreatment with wortmannin, a PI3K/Akt pathway inhibitor, significantly inhibited p-Akt expression and further attenuated both transcription and translation of HIF 1alpha and VEGF. Collectively, our findings suggested that PI3K/Akt signaling pathway might be involved in HIF-1alpha regulation after OGD in cultured cortical neurons. PMID- 19553018 TI - Momentary pain and coping in temporomandibular disorder pain: exploring mechanisms of cognitive behavioral treatment for chronic pain. AB - The purpose of this study was to determine whether cognitive-behavioral treatment (CBT) operates by effecting changes in cognitions, affects, and coping behaviors in the context of painful episodes. Patients were 54 men and women with temporomandibular dysfunction-related orofacial pain (TMD) enrolled in a study of brief (6 weeks) standard conservative treatment (STD) or standard treatment plus CBT (STD+CBT). Momentary affects, pain, and coping processes were recorded on a cell phone keypad four times per day for 7 days prior to treatment, and for 14 days after treatment had finished, in an experience sampling paradigm. Analyses indicated no treatment effects on general retrospective measures of pain, depression, or pain-related interference with lifestyle at post-treatment. However, mixed model analyses on momentary pain and coping recorded pre- and post treatment indicated that STD+CBT patients reported greater decreases in pain than did STD patients, significantly greater increases in the use of active cognitive and behavioral coping, and significantly decreased catastrophization. Analyses of experience sampling data indicated that post-treatment momentary pain was negatively predicted by concurrent active coping, self-efficacy, perceived control over pain, and positive-high arousal affect. Concurrent catastrophization was strongly predictive of pain. Active behavioral coping and self-efficacy reported at the prior time point (about 3h previously) were also protective, while prior catastrophization and negative-high arousal mood were predictive of momentary pain. The results suggest that CB treatment for TMD pain can help patients alter their coping behaviors, and that these changes translate into improved outcomes. PMID- 19553019 TI - Complementary techniques for the characterization of thin film Ti/Nb multilayers. AB - An aberration corrector on the probe-forming lens of a scanning TEM (STEM) equipped with an electron energy-loss spectrometer (EELS) and X-ray energy dispersive spectrometer (XEDS) has been employed to investigate the compositional variations as a function of length scale in nanoscale Ti/Nb metallic multilayers. The composition profiles of EELS and XEDS were compared with the profiles obtained from the complementary technique of 3D atom probe tomography. At large layer widths (h > or = 7 nm, where h is the layer width) of Ti and Nb, XEDS composition profiles of Ti/Nb metallic multilayers are in good agreement with the EELS results. However, at reduced layer widths (h approximately 2 nm), profiles of EELS and atom probe exhibited similar compositional variations, whereas XEDS results have shown a marked difference. This difference in the composition profiling of the layers has been addressed with reference to the effects of beam broadening and the origin of the signals collected in these techniques. The advantage of using EELS over XEDS for these nanoscaled multilayered materials is demonstrated. PMID- 19553020 TI - Characteristics and outcomes of multiple organ dysfunction syndrome among severe burn patients. AB - AIM: Determining the characteristics and outcomes of multiple organ dysfunction syndrome (MODS) in severe-burn patients. METHOD: A prospective study was conducted in 117 severe-burn, adult patients admitted to the National Institute of Burns, Hanoi, Vietnam with burn area > or =40% of the total body surface area. The diagnosis of MODS was based on Sequential Organ Failure Assessment (SOFA) score. RESULTS: MODS was recorded in 45.3% of the patients. A higher rate of MODS was recorded in patients over 40 years of age (51.61%), those presenting with inhalation injury (60.37%) and having a large burn surface area. MODS was commonly seen in the second week after-burn (75.47%). Respiratory system failure was the most common (44.44%), followed by circulatory system failure (41.88%) and failure of other systems. MODS was more common among patients developing sepsis and septic shock (69.64% and 87.5%, respectively). The mortality rate was 86.79% among MODS patients and higher in case of SOFA score > or =6. In addition, mortality rate was 22.22% if one organ was involved, 40% for two organs, 93.33% for three organs and 100% if four or more organs were involved. The durations of artificial ventilation, hospitalisation and intensive care unit stay were significant higher than in MODS patients as compared to non-MODS patients. CONCLUSION: MODS is still a severe complication, leading to death after-burn. It is important to identify the risk factors and prevention methods to increase the chances of saving severely burned patients. PMID- 19553021 TI - Does "off-hours" admission affect burn patient outcome? AB - INTRODUCTION: Previous critical care and cardiology studies find that critically ill patients have worse outcomes when admitted to the hospital during off-hours as compared to those admitted during weekdays. As severe burn is equally emergent we hypothesized that this disparity in outcomes would exist for burn patients as well. STUDY DESIGN: Secondary analysis of the National Trauma Data Bank (NTDB) version 7.1. The NTDB is a national registry of hospital admissions for traumatic injury administered by the American College of Surgeons. SETTING: 700 trauma facilities nationwide contributing to the NTDB between 2002 and 2006. SUBJECTS: All trauma patients included in the dataset with the injury mechanism of burn divided into "off-hours" admits (nights from 6pm to 6am and weekends) and weekday admits. MEASURES: Time and day of admission, demographics, ISS score, injury characteristics (+/-inhalational injury, TBSA, and full thickness injury), facility characteristics (number of burn beds, teaching status). OUTCOMES: Mortality as the primary outcome. Secondary outcomes include ICU length of stay (LOS), hospital length of stay. DATA ANALYSIS: Descriptive statistics to summarize group characteristics, chi(2) and Student's t tests for bivariate analysis, multivariable linear and logistic regressions. RESULTS: Of the 25,572 burn patients, 17,625 (68.9%) arrived during off-hours. There was no difference in ICU length of stay (LOS) (p=0.233), hospital LOS (p=0.82), or mortality (p=0.546) for those admitted during off-hours compared with weekday admits. In multivariate analysis when controlling for age, gender, burn characteristics (inhalation injury, full thickness injury, and TBSA >30%), and hospital type, off hours admission was not predictive of mortality (OR=1.06, 95% CI 0.91-1.23). CONCLUSIONS: Contrary to studies in other critically ill patient populations, off hours admission is not predictive of worse outcomes in burn patients. PMID- 19553022 TI - Fatal rescue burns--possible theories of causation. PMID- 19553023 TI - Garlic burn: the real facts. PMID- 19553024 TI - Twelve years epidemiological study of paediatric burns in Ain Shams University, Burn Unit, Cairo, Egypt. PMID- 19553025 TI - A small-area population analysis of socioeconomic status and incidence of severe burn/fire-related injury in British Columbia, Canada. AB - Socioeconomic determinants of injury have been associated with risk of burn in the UK and USA, but the relative significance of this impact is largely unknown across Canadian populations. The purpose of this study is to determine the extent to which socioeconomic status (SES) is linked to risk of burn in the province of British Columbia (BC) and identify the extent to which these findings are generalizable across both urban and rural population groups. Measures of SES were based on province-wide comparisons using data obtained from the Canada Census using the Vancouver Area Neighbourhood Deprivation Index (VANDIX). Results illustrate that the effects of SES and increased injury risk are substantial, though the most pronounced variations were exhibited across each SES stratum for urban areas and with less demonstrable effect when itemized by injury type within rural areas. Although conservative, the results from this study illustrate that burns disproportionately affect populations of greater relative socioeconomic disadvantage and continued efforts to also address social inequities and their link to injury incidence is likely to be more effective than targeting individual behavior alone when trying to reduce and eliminate their occurrence. PMID- 19553026 TI - Sex-dependent alterations in response to maternal deprivation in rats. AB - We review here our latest results regarding short- and long-term effects of a neonatal maternal deprivation (MD) stress [24h at postnatal day (PND) 9] on diverse psychoneuroimmunoendocrine parameters, pointing out the existence of numerous sexual dimorphisms. Behavioral changes observed in MD animals might be at least in part attributable to neurodevelopmental effects of MD-induced elevated corticosterone levels. Our findings of short-term effects of MD on hippocampal and cerebellar neurons and glial cells appear to support this hypothesis. However, it is important to note that these cellular effects were more marked in males than in females. Moreover, in analyzing the effects of this neonatal stress on the endocannabinoid system (hippocampal endocannabinoid levels and CB1 receptors) we have also found that males were more affected by MD. Since all these sexual dimorphisms were found at an early neonatal age (PND 13), they are attributable to organizational effects of gonadal steroids. We discuss the potential implications of the elevated corticosterone and decreased leptin levels shown by MD animals in their diverse functional alterations, including the above mentioned neural effects as well as the intriguing persistent deficit in their immunological system. We also emphasize the necessity of analyzing the important influence of sex as regards the specific consequences of early life stress. PMID- 19553028 TI - Myosin heavy chain proteins are responsible for the development of obstructive sleep apnea syndrome. AB - We introduce a hypothesis that obstructive sleep apnea syndrome (OSAS) is primarily caused by an inherited reduced adaptability of upper airway striated muscles such that they cannot maintain patency when there is reduced consciousness (sleep). This reduced ability is caused by a deficiency of the genes for specific myosin heavy chain (MHC) proteins, which are the primary source of muscle adaptability in adults and were initially described in the chewing muscles. The development of OSAS must be linked to problems with striated muscle because affected patients are capable of normal breathing when awake but their respiratory parameters deteriorate during sleep; OSAS must, therefore, be caused by a factor that is voluntarily active during waking but inactive during sleep, and this can only be striated muscle. Congenital or acquired anatomical abnormalities are involved only partially, because OSAS patients with anatomical abnormalities do not begin to snore or to have apneas or hypopneas when lying in bed awake, but begin to do so only when sleeping. PMID- 19553029 TI - Therapeutic ultrasound for dental tissue repair. AB - Dental disease affects human health and the quality of life of millions worldwide. Tooth decay (caries) and diseases of the dental pulp result in loss of tooth vitality and function requiring invasive treatment to restore the tooth to health. "Therapeutic" low intensity pulsed ultrasound has been shown to accelerate bone fracture healing indicating that ultrasound may be used as a tool to facilitate hard tissue regeneration. We have shown recently that low frequency ultrasound is able to exert biological effects on odontoblast-like cells. In this paper, we postulate that low frequency, low intensity ultrasound may stimulate endogenous coronal tooth repair by stimulating dentine formation from existing odontoblasts or by activating dental pulp stem cells to differentiate into new reparative dentine-producing cells. Ultrasound therapy promoting dentine formation and repair may also have the potential benefit of alleviating dentine hypersensitivity by inducing occlusion of dentinal tubules. It is envisaged that therapeutic ultrasound may be used in future to facilitate dental tissue engineering and stem cell therapy applications for dental tissue regeneration. Further research is warranted in this clinically important area and we envisage that novel strategies in dental therapy will be realised that may ultimately lead to the development of novel non-invasive, multifunctional ultrasound devices for dental diagnostics, repair and regeneration. PMID- 19553027 TI - Oxytocin protects against negative behavioral and autonomic consequences of long term social isolation. AB - Positive social interactions and social support may protect against various forms of mental and physical illness, although the mechanisms for these effects are not well identified. The socially monogamous prairie vole, which--like humans--forms social bonds and displays high levels of parasympathetic activity, has provided a useful model for investigating neurobiological systems that mediate the consequences of sociality. In the present study, adult female prairie voles were exposed to social isolation or continued pairing with a female sibling (control conditions) for 4 weeks. During weeks 3 and 4 of this period, animals were administered oxytocin (20 microg/50 microl, s.c.) or saline vehicle (50 microl, s.c.) daily for a total of 14 days. In Experiment 1, autonomic parameters were recorded during and following isolation or pairing. Isolation (vs. pairing) significantly increased basal heart rate (HR) and reduced HR variability and vagal regulation of the heart; these changes in isolated animals were prevented with oxytocin administration. In Experiment 2, behaviors relevant to depression [sucrose intake and swimming in the forced swim test (FST)] were measured as a function of isolation. Isolation reduced sucrose intake and increased immobility in the FST; these behaviors also were prevented by oxytocin. Administration of oxytocin did not significantly alter cardiac, autonomic or behavioral responses of paired animals. These findings support the hypothesis that oxytocinergic mechanisms can protect against behavioral and cardiac dysfunction in response to chronic social stressors, and can provide insight into social influences on behavior and autonomic function in humans. PMID- 19553030 TI - Inadvertent potential risk of neoadjuvant chemotherapy in cervical cancer. AB - Randomized clinical trials comparing neoadjuvant chemotherapy (NAC) followed by surgery with initial surgery in cervical cancer reported conflicting results. We provided a hypothesis explaining the different outcomes between trials. NAC was reported to make a lymph node metastasis smaller. Smaller tumors are hard to detect and potentially undetected at pathologic examination. Undetected metastasis would lead to an erroneous exclusion of high-risk patients from adjuvant therapy. An erroneous exclusion would lead to a recurrence. This harmful concealing effect of NAC could negate the potentially beneficial effect of NAC and the conflicting results of trials could be the result of the balance between harmful and beneficial effect of NAC. Because the concealing effect of NAC can be reversed by adjuvant therapy, trials comparing NAC followed by surgery with initial surgery in which all patients in both arms received adjuvant therapy reported positive result. However, in a trial in which only patients with lymph node or parametrial involvements received adjuvant therapy, the concealing effect was not reversed by adjuvant therapy and reported negative result. Our empirical data showed that patients who underwent NAC followed by surgery and were classified as low-risk based on pathologic parameters had unexpectedly high recurrence rate. To evaluate our hypothesis, a trial comparing a conventional pathologic examination with a more sophisticated examination such as ultrastaging in patients with cervical cancer who received NAC followed by surgery would be necessary. If our hypothesis is proven, trials considering the concealing effect of NAC should be performed to accurately evaluate the role of NAC in cervical cancer. PMID- 19553031 TI - Whole-pelvis radiotherapy in combination with interstitial brachytherapy: does coverage of the pelvic lymph nodes improve treatment outcome in high-risk prostate cancer? AB - PURPOSE: To compare biochemical progression-free survival (bPFS), cause-specific survival (CSS), and overall survival (OS) rates among high-risk prostate cancer patients treated with brachytherapy and supplemental external beam radiation (EBRT) using either a mini-pelvis (MP) or a whole-pelvis (WP) field. METHODS AND MATERIALS: From May 1995 to October 2005, 186 high-risk prostate cancer patients were treated with brachytherapy and EBRT with or without androgen-deprivation therapy (ADT). High-risk prostate cancer was defined as a Gleason score of > or =8 and/or a prostate-specific antigen (PSA) concentration of > or =20 ng/ml. RESULTS: With a median follow-up of 6.7 years, the 10-year bPFS, CSS, and OS rates for the WP vs. the MP arms were 91.7% vs. 84.4% (p = 0.126), 95.5% vs. 92.6% (p = 0.515), and 79.5% vs. 67.1% (p = 0.721), respectively. Among those patients who received ADT, the 10-year bPFS, CSS, and OS rates for the WP vs. the MP arms were 93.6% vs. 90.1% (p = 0.413), 94.2% vs. 96.0% (p = 0.927), and 73.7% vs. 70.2% (p = 0.030), respectively. Among those patients who did not receive ADT, the 10-year bPFS, CSS, and OS rates for the WP vs. the MP arms were 82.4% vs. 75.0% (p = 0.639), 100% vs. 88% (p = 0.198), and 87.5% vs. 58.8% (p = 0.030), respectively. Based on multivariate analysis, none of the evaluated parameters predicted for CSS, while bPFS was best predicted by ADT and percent positive biopsy results. OS was best predicted by age and percent positive biopsy results. CONCLUSIONS: For high-risk prostate cancer patients receiving brachytherapy, there is a nonsignificant trend toward improved bPFS, CSS, and OS rates when brachytherapy is given with WPRT. This trend is most apparent among ADT-naive patients, for whom a significant improvement in OS was observed. PMID- 19553032 TI - Seed implant retention score predicts the risk of prolonged urinary retention after prostate brachytherapy. AB - PURPOSE: To risk-stratify patients for urinary retention after prostate brachytherapy according to a novel seed implant retention score (SIRS). PATIENTS AND METHODS: A total of 835 patients underwent transperineal prostate seed implant from March 1993 to January 2007; 197 patients had (125)I and 638 patients had (103)Pd brachytherapy. Four hundred ninety-four patients had supplemental external-beam radiation. The final downsized prostate volume was used for the 424 patients who had neoadjuvant hormone therapy. Retention was defined as reinsertion of a Foley catheter after the implant. RESULTS: Retention developed in 7.4% of patients, with an average duration of 6.7 weeks. On univariate analysis, implant without supplemental external-beam radiation (10% vs. 5.6%; p = 0.02), neoadjuvant hormone therapy (9.4% vs. 5.4%; p = 0.02), baseline alpha blocker use (12.5% vs. 6.3%; p = 0.008), and increased prostate volume (13.4% vs. 6.9% vs. 2.9%, >45 cm(3), 25-45 cm(3), <25 cm(3); p = 0.0008) were significantly correlated with increased rates of retention. On multivariate analysis, implant without supplemental external-beam radiation, neoadjuvant hormone therapy, baseline alpha-blocker use, and increased prostate volume were correlated with retention. A novel SIRS was modeled as the combined score of these factors, ranging from 0 to 5. There was a significant correlation between the SIRS and retention (p < 0.0001). The rates of retention were 0, 4%, 5.6%, 9%, 20.9%, and 36.4% for SIRS of 0 to 5, respectively. CONCLUSIONS: The SIRS may identify patients who are at high risk for prolonged retention after prostate brachytherapy. A prospective validation study of the SIRS is planned. PMID- 19553033 TI - Dysphagia after chemoradiotherapy for head-and-neck squamous cell carcinoma: dose effect relationships for the swallowing structures. AB - PURPOSE: To evaluate late dysphagia after chemoradiotherapy for locally advanced head-and-neck squamous cell carcinoma, and to examine its correlation with clinical and dosimetric parameters. METHODS AND MATERIALS: Consecutive patients, treated with radiotherapy (70-72 Gy) and concomitant chemotherapy (cisplatinum 100 mg/m(2) every 3 weeks) between 2004 and 2007, were examined. Swallowing was evaluated by four quality-of-life questionnaires: EORTC C30 and H&N35, the Performance Status Scale of List, and the MD Anderson Dysphagia Inventory. Clinical and dosimetric parameters were correlated with late dysphagia. RESULTS: A total of 53 disease-free patients were evaluated; mean follow-up was 20.4 months (range, 6-45 months). The volume of the middle pharyngeal constrictor muscle receiving > or =50 Gy (p = 0.04), the mean dose to this structure (p = 0.02) and to the supraglottic larynx (p = 0.04) were significantly associated with late swallowing problems at univariate analysis, along with tumor localization (p = 0.008), T-classification (p = 0.02), and pretreatment swallowing problems (p = 0.01). Only this last factor significantly correlated with late dysphagia at multivariate analysis. CONCLUSION: These findings motivate further efforts to reduce the dose to the swallowing structures, especially to the pharyngeal constrictor muscles and the larynx. However, clinical parameters are also important and should be included in future prospective trials. PMID- 19553034 TI - Selective nodal irradiation for head and neck cancer using intensity-modulated radiotherapy: application of RTOG consensus guidelines in routine clinical practice. AB - PURPOSE: We have been using intensity-modulated radiotherapy (IMRT) for selective neck irradiation. This article presents an analysis of patterns of failure and their dosimetric correlation. METHODS AND MATERIALS: Between October 2003 and January 2008, 83 patients with head-and-neck cancer were treated with IMRT. Nodal levels were contoured as per the Radiation Therapy Oncology Group (RTOG) consensus guidelines. RESULTS: There were 32 relapses with 23 local relapses (21 local relapses alone and 2 local and regional relapses, simultaneously), 9 regional relapses (including 2 simultaneous local and regional relapses), and 5 distant relapses, of which 2 patients had local relapses. At 2 and 3 years, the locoregional relapse-free survival rates were was 68.3% and 60.8%, respectively, while the overall survival rates were 84.1% and 81.7%, respectively. Subgroup analyses revealed significant differences in locoregional relapse-free survival rates for total treatment times of <53 days vs. >53 days, a volume of CTV1PTV (i.e., the volume prescribed 70 Gy) <177 cc vs. >177 cc, a V100 for CTV1PTV of <91% vs. >91%, and a minimum dose to CTV1PTV of <54 Gy vs. >54 Gy. There were no failures in the elective nodal volume, substantiating both the nodal selection criteria and the RTOG consensus guidelines for delineation of neck node levels. CONCLUSIONS: IMRT for head-neck cancer is feasible, using elective nodal selection criteria along with RTOG consensus guidelines for the radiological boundaries of levels of neck nodes. PMID- 19553035 TI - PCB126 enhanced the genotoxicity of BaP in HepG2 cells by modulating metabolic enzyme and DNA repair activities. AB - Both of benzo(a)pyrene (BaP) and 3,3',4,4',5-pentachlorobiphenyl (PCB126) are ubiquitous and persistent environmental pollutants. These two chemicals coexist in various environmental media and human samples and thus may have combined effects on human health. However, the toxic effects and related mechanism of co exposure to BaP and PCB126 remain unknown. In a series of experiments using the HepG2 cells exposed to BaP (50microM) or/and PCB126 (0.01, 0.1, 1 and 10nM), we measured the rate of micronucleus (MN) formation, CYP1A1 activity and expression of nucleotide excision repair (NER) proteins (XPA and XPC). We found that the exposure to BaP or PCB126 alone could effectively increase the CYP1A1 activity and the XPA expression. BaP alone had a profound enhancement of MN formation. Compared with BaP alone, co-exposure to both BaP and PCB126 significantly enhanced the CYP1A1 activity and the formation of MN but reduced the expression of both XPA and XPC. The synergistic effect of PCB126 on BaP-induced MN formation was inhibited by alpha-naphthoflavone (ANF), an inhibitor of CYP1A1. Our findings suggest that PCB126 may enhance BaP-induced DNA damage and genotoxicity by increasing cytochrome P450 1A activity and decreasing the NER capacity. PMID- 19553036 TI - Localization of thyrotropin receptor and thyroglobulin in the bovine corpus luteum. AB - The receptor of the Thyroid Stimulating Hormone (TSHr) and thyroglobin (TGB), are two proteic factors necessary for the synthesis of hormones, in the thyrocite. In mammals, many immuno-histochemical reports indicate the presence of the TSHr in extra-thyroidal tissues, but not in the ovary. Triiodothyronine (T(3)) and thyroxine (T(4)) have been widely shown to affect ovarian functions and the synthesis of progesterone (P(4)). The aim of this study was to determine if by immunohistochemistry techniques TSHr and TGB could be found in the bovine corpora haemorragica, lutea and albicantia. A primary rabbit polyclonal antibody against human TSHr and a primary rabbit polyclonal antibody against human TGB were employed. Furthermore, the accuracy of bovine thyroid to the antibodies used in this study was tested. A positivity reaction for the anti-TSHr serum in the large luteal cells and immunostaining of both small and large luteal cells with the anti-TGB serum occurred only in mature corpora lutea. No immunostaining was detected in stromal cells, blood and lymphatic vessels and in corpora haemorragica and albicantia. Bovine thyroid tissue showed immunostaining to both the antibodies employed. These data suggest that the luteal cells of mature corpora lutea may be involved in the synthesis of thyroid hormones, which may modulate P(4) synthesis, acting in an autocrine and paracrine way. PMID- 19553037 TI - Osmotic stress stimulates generation of superoxide anion by spermatozoa in horses. AB - The objective of this study was to examine the interplay between osmotic and oxidative stress as well as to determine mechanisms by which osmotic stress increases superoxide generation in spermatozoa of horses. Superoxide production, as measured by dihydroethidium (DHE), increased when spermatozoa of horses were incubated under either hyperosmotic or hyposmotic conditions. This increase in superoxide production was inhibited by the MAP kinase p38 inhibitor, SB203580, and by the superoxide scavenger, tiron. Incubation of spermatozoa under hyperosmotic conditions increased overall protein tyrosine phosphorylation as measured by western blotting techniques; however, a similar increase was not detected when spermatozoa were incubated under hyposmotic conditions. The general protein kinase C (PKC) and protein tyrosine kinase (PTK) inhibitor staurosporine inhibited (P<0.05) tyrosine phosphorylation in samples from cells under hyperosmotic conditions. In addition, the NADPH oxidase inhibitor diphenyleneiodonium (DPI) also inhibited (P<0.05) protein tyrosine phosphorylation in cells under hyperosmotic conditions. In summary, these data indicate that incubation of equine spermatozoa under both hyposmotic and hyperosmotic conditions can increase superoxide anion generation. Under hyperosmotic conditions, this increased generation of superoxide anion was accompanied by increased protein tyrosine phosphorylation. PMID- 19553038 TI - Effect of the bilateral hip bone density measurement on clinical practice in elderly subjects. AB - OBJECTIVES: The aim of this study was to determine whether the addition of the measurement of bilateral hip bone mineral density (BMD) has an impact on indications for osteoporosis (OP) treatment in community-dwelling elderly individuals, based on criteria from the National Osteoporosis Foundation (NOF). METHODS: In total, 605 consecutive community-dwelling elderly individuals who were 65 years and older were evaluated. Dual energy X-ray absorptiometry was used to determine the lowest T-score in the lumbar spine+unilateral hip, the bilateral hips, and the lumbar spine+bilateral hips. Risk factors associated with the lowest T-score in these three conditions were applied to indicate treatment in accordance with NOF criteria. McNemar's test was used to assess the difference of adding bilateral hip BMD measurements. RESULTS: There was a significant difference in the frequency of pharmacological indication using NOF criteria together with the lowest T-score for the three tests (72.8% for lumbar spine+bilateral hips and 71.2% for lumbar spine+unilateral hip; p=0.002). A higher frequency of treatment indication was also observed for lumbar spine+unilateral hip (71.2%) compared to bilateral hips (61.1%) (p<0.001). The discrepancies in treatment appeared to be more evident in women when analyzed by gender distribution. CONCLUSION: Our finding supports the theory that evaluation of the bilateral hips with the lumbar spine seems to be more sensitive measure for identifying patients with an osteoporosis treatment indication. Furthermore, despite the well-known artifact in the lumbar spine, this site should not be excluded when determining the indication for OP treatment in elderly people. PMID- 19553039 TI - Clinicopathologic significance of CD105-assessed microvessel density in glottic laryngeal squamous cell carcinoma. AB - OBJECTIVE: Intratumoral microvessel density (MVD) determined with the use of antibodies to endoglin (CD105) is considered to be an important prognostic marker in a variety of malignancies. The purpose of this study has been to analyze the clinicopathologic significance of CD105-assessed MVD in SCCs primary localized in glottic region of larynx. METHODS: Surgical specimens from 40 patients with resected glottic squamous cell carcinomas were immunostained for CD105. CD105 assessed MVD was calculated at 400x magnification. Using the mean MVD as a cut off, tumors were classified in the "high MVD" group and in the "low MVD" group. Clinicopathologic data were collected retrospectively. RESULTS: The mean MVD assessed by CD105 in considered glottic SCCs was 12.3 (standard deviation [SD]=3.65). MVD varied among tissue samples from 5 to 21 (median 12.5). High MVD was significantly correlated with a more aggressive tumor phenotype, including T3 T4 tumor (Fisher exact test, P=0.004) and advanced clinical stage (Fisher exact test P=0.026). Kruskal-Wallis test identified significant relation between pT stages and CD105-assessed MVD (P=0.011). CD105-assessed MVD was significantly related to malignancy recurrence presence/absence (Mann-Whitney U-test P=0.023). Logistic regression in multivariate modality showed that MVD (odds ratio [OR] 2.29, P=0.033, 95% confidence interval [CI] 1.06-7.53) and advanced T category (T3-T4) (OR 4.11, P=0.026, 95% CI 2.38-9.46) were significantly related to malignancy recurrence presence/absence. Cox regression analysis revealed that expression of CD105 (P=0.031) and N status (P=0.014) were the independent factors for disease-free survival. CONCLUSION: High expression of CD105 correlated significantly with advanced T status and locoregional recurrence. The present preliminary results suggest that CD105-assessed MVD in primary glottic squamous cell carcinomas may identify patients at risk of disease recurrence. PMID- 19553040 TI - Laryngeal kaposiform hemangioendothelioma: case report and literature review. AB - The kaposiform hemangioendothelioma (KHE) is an uncommon vascular neoplasm of borderline or intermediate malignant potential in which involvement of the head and neck region is rare. A 5-year-old girl was admitted to the emergency room with abrupt massive hemoptysis. Transoral laryngoscopy revealed a round, reddish mass with active bleeding on the anterior portion of the left vocal fold region. Since re-hemoptysis and large amount of aspiration caused breathing difficulties for the patient, suspension exam was performed under general anesthesia. Mass excision and thorough bleeding control was performed with laser assisted laryngo micro system. Postoperatively, there was no further bleeding. The pathologic diagnosis of the tumor was KHE. To the best of our knowledge, this is the first report of a KHE in the larynx presenting as recurrent hemoptysis in childhood. PMID- 19553041 TI - Kurz titanium prosthesis ossiculoplasty--follow-up statistical analysis of factors affecting one year hearing results. AB - OBJECTIVE: Analysis of the one year results of Kurz titanium ossicular prosthesis and the factors affecting the outcome from this prosthesis. The hearing results of titanium partial ossicular replacement prosthesis (PORP) and total ossicular replacement prosthesis (TORP) were compared. METHODS: This is prospective study of 97 Kurz prosthesis ossiculoplasties, performed by the same senior author (VVR) between 2004 and 2006. All patients undergoing ossiculoplasty using Kurz prosthesis between 2004 and 2006 were included. All patients had minimum follow up period of one year. There were 97 patients in total. 65 patients had PORP's and 32 had TORP's. The mean preoperative and postoperative air-bone gaps for the frequencies (500, 1000, 2000, and 3000Hz) were calculated. The improvement of the mean air-bone gap (ABG) and air conduction over the same frequencies were measured. A postoperative ABG less than or equal to 20dB was considered a successful result. Statistical analysis was used to identify the factors which affect the postoperative results. RESULTS: The mean preoperative ABG was 27.89dB (SD=11.71). The mean postoperative ABG for the entire series was 11.39dB (SD=10.47). The mean improvement in ABG was 16.50dB (SD=14.00). 81.4% (79 patients) had postoperative ABG< or =20dB. 56.7% (55 patients) of the patients had postoperative ABG< or =10dB. The mean postoperative ABG for all PORP patients was 10.6dB (SD=9.7). The mean postoperative ABG of TORP was higher in this study (14.84dB, SD=12.86) but it was not statistically significant (p=0.10, 2-tailed t test, 95% CI -9.35 to 0.924). There was no significant effect of age, presence or absence of cholesteatoma or retraction pocket, type of mastoid surgery and thickness of the cartilage graft used on the results. The effect of the preoperative ossicular condition on the postoperative hearing results was also analysed. CONCLUSION: Titanium ossicular reconstruction gives stable short-term results. There was no statistically significant difference between the total and partial replacement prostheses. The preoperative status of the stapes superstructure did influence the mean postoperative ABG. PMID- 19553042 TI - New treatment for invasive fungal sinusitis: three cases of chronic invasive fungal sinusitis treated with surgery and voriconazole. AB - Invasive fungal sinusitis is a relatively rare disease and can be divided into acute fulminant, chronic, and granulomatous invasive fungal sinusitis. The conventional treatment is radical surgery combined with systemic amphotericin B administration, but the poor prognosis and unestablished treatment options require a better therapeutic strategy. We report three cases of chronic invasive fungal sinusitis successfully treated with a combination of surgery and voriconazole, a new antifungal agent, with good responses in all patients. Voriconazole administration could form the basis for a new standard treatment for invasive fungal sinusitis. PMID- 19553043 TI - Study of the human hypoglossal nucleus: normal development and morpho-functional alterations in sudden unexplained late fetal and infant death. AB - This study evaluated the development and the involvement in sudden perinatal and infant death of the medullary hypoglossal nucleus, a nucleus that, besides to coordinate swallowing, chewing and vocalization, takes part in inspiration. Through histological, morphometrical and immunohistochemical methods in 65 cases of perinatal and infant victims (29 stillbirths, 7 newborns and 29 infants), who died of both unknown and known cause, the authors observed developmental anomalies of the hypoglossal nucleus (HGN) in high percentage of sudden unexplained fetal and infant deaths. In particular, HGN hypoplasia, hyperplasia, positive expression of somatostatin and absence of interneurons were frequently found particularly in infant deaths, with a significant correlation with maternal smoking. PMID- 19553045 TI - [Occurrence of lead poisoning during follow-up of children at risk with initial screening lead blood levels below 100 microg/L]. AB - BACKGROUND: Follow-up is recommended for children initially screened with a lead blood level below the threshold for lead poisoning of 10 microg/dL when they have one or more risk factors. At first, the frequency of a follow-up lead blood test was calculated in children at risk for lead poisoning. In second time, we calculated the rate of occurrence and independent factors for lead poisoning in the follow-up group. METHODS: Since 1992, the Greater Paris lead poisoning monitoring system (SSSIILF) has been systematically recording data on lead levels in blood tests conducted for screening and follow-up in Greater Paris. Children initially screened before the age of seven whose blood lead level was inferior to 10 microg/dL and who had one or more risk factors were selected. The association between qualitative variables and a follow-up lead blood test was compared using the Chi(2) test. For children given follow-up, the association between qualitative variables and occurrence of lead poisoning was compared using the Chi(2) test; independent factors for lead poisoning were estimated by logistic regression. RESULTS: A follow-up lead blood test was more frequent and the difference was statistically significant, for children with one or more of the following risk factors identified at the time of screening: home address in Seine Saint-Denis or central Paris, screened in mother/child healthcare centers (PMI) or through a private physician, a blood lead level 5 microg/dL on initial screening, young age (<24 months) at the time of screening and some others known risk factors. The rate of occurrence of lead poisoning during follow-up was 25.9% for children screened between 1992 to 1994 and decrease to 5.1% for children screened in 2004 to 2005 (p<0.001) and was lower in central Paris and Seine Saint Denis than in other districts in Greater Paris (p<0.01). The rate of occurrence during follow-up, independent of known risk factors for lead poisoning (p<0.01), was higher for children screened before the age of two (p<0.01) and for children whose mothers were from Sub-Saharan Africa (p<0.01). CONCLUSION: It is essential to follow up children at risk with an initial lead blood level below 10 microg/dL, especially those initially screened before the age of 24 months. Local action on home environment could also be needed when the initial blood lead level is 5 microg/dL and more than one risk factor has been identified. PMID- 19553046 TI - [Tobacco consumption among young adults in the two French departments of Savoie in 2008]. AB - BACKGROUND: As a result of tobacco control measures in France, smoking among adolescents is decreasing. However, this decrease is associated with changes in the way youth are consuming tobacco and turning towards new tobacco products: cheaper forms of tobacco such as rolling or chewing tobacco, or fashionable forms such as narghile. The aim of this study is to describe in young adults: (1) prevalence of tobacco use and main risk factors of daily smoking, (2) the entry mode for tobacco use and (3) prevalence and main risk factors related to "chewing tobacco" used as snuff. METHODS: A descriptive transversal study was undertaken in five private and public high schools in the French Alps region in 2008. Anonymous questionnaires were given out to the students of 12th grade (last year of general secondary education) and BTS (professional training). Smoking prevalence and other forms of tobacco consumption were described, as well as the entry mode in tobacco use. Finally, we used logistic models to identify the main determinants of smoking cigarettes and using chewing tobacco. RESULTS: This study included 920 students: 22.3% (95% IC: 19.6-25.0) were daily smokers and 65.9% (95% IC: 62.8-69.0) had tried tobacco. Approximately 40% had experimented with rolling tobacco, cannabis or narghile. We found the usual determinants of daily smoking: an environment conducive to smoking, and not belonging to a sports club. Around 11% (95% IC: 9.2-13.2) had tried chewing tobacco. Risk factors associated with chewing tobacco were: having a smoking friend (adjusted OR: 3.07; 95% IC: 1.95-4.83), studying in a private school (adjusted OR: 2.57; 95% IC: 1.52-4.31), or being male (adjusted OR: 1.79; 95% IC: 1.15-2.79). CONCLUSION: As found in national studies, cigarette smoking is declining among young adults, but the use of other tobacco products (narghile, chewing tobacco) is emerging. The relatively high consumption rate and the risk factors of chewing tobacco need to be examined in depth in order to organize prevention programs for young adults. PMID- 19553047 TI - [Hepatic surgery. What progress? What future?]. AB - Progress in liver surgery has increased the safety of all types of anatomical liver resections and has, in association with chemotherapy and interventional radiology, extended the limits of resectability for many primary and secondary liver tumors. This progress has been achieved thanks to a better method of vascular clamping, vascular reconstruction and better techniques and instruments to achieve more precise liver transection of this extremely vascular organ with a good haemostasis on the cut surface. These technical advances have allowed the procurement of partial liver grafts from living donors for transplantation and have also extended the scope of laparoscopic access to practically all types of hepatic resections. Preoperative manipulation of the liver volume with hypertrophy of the future liver remnant has made complex and extended hepatectomies feasible. These major hepatic resections are often performed in the background of damaged liver parenchyma due to chronic liver disease, steatosis and chemotherapy-induced lesions. A better knowledge of the functional anatomy with a clearer understanding of the role of venous drainage in the recovery of liver function and the process of regeneration have been the major factors which have facilitated these extended resections. The future of hepatic surgery, which is constantly influenced by the continuous technical progress with computer-aided reconstruction of the tumor zones and vascular and biliary anatomy, is certainly going to see a robotic-assisted surgical approach. Indications will change. While major and complex resections will still be required for advanced tumors, very few benign lesions will be operated in the future. Huge changes will happen in the management of malignant lesions where surgery will become a part of a combined therapeutic strategy where oncologic and radiointerventional treatments will have a major role. Surgery could then be limited to the resection of residual nodules susceptible to harbour persistent viable tumor cells. PMID- 19553048 TI - Diffusion-weighted magnetic resonance imaging in differentiation of postobstructive consolidation from central lung carcinoma. AB - PURPOSE: To prospectively evaluate diffusion-weighted (DW) magnetic resonance (MR) imaging for differentiation of postobstructive consolidation from centrally located lung carcinomas by using apparent diffusion coefficients (ADCs). MATERIALS AND METHODS: An institutional review board approved this study; informed consent was obtained from patients. Forty-nine consecutive patients (3 women, 46 men; mean age, 63.6 years; age range, 42-85 years) with lung carcinoma underwent DW MR imaging. Forty patients had central and nine patients had peripheral lung carcinomas. ADC of each lung carcinoma was calculated from DW MR images obtained with two different b values (0, 1000 s/mm(2)). In the final study group including 27 patients with central lung carcinoma accompanying distal lung consolidation (mean age, 67.2 years; 3 women, 24 men), ADCs of lung carcinomas were statistically compared among cytologic/histologic types and accompanying postobstructive consolidations. Unpaired t test was used for measurable variables with normal distribution, and Kruskal-Wallis variance analysis and Mann-Whitney U tests were used for the measurable variables without normal distribution. RESULTS: There was no significant difference between mean ADC values of all types of carcinomas (P=.302) and also between mean ADC values of central (1.91 +/- 0.7x10(-3) mm(2)/s) and peripheral carcinomas (1.58 +/- 0. 6x10(-3) mm(2)/s) (P=.224). The mean ADC value for the masses of central lung carcinoma with postobstructive consolidations was 1.83 +/- 0.75x10(-3) mm(2)/s, and for consolidation was 2.50 +/- 0.76x10(-3) mm(2)/s. ADC of central carcinoma masses was significantly lower than that of postobstructive consolidations (P=.003). CONCLUSIONS: ADC values of central lung carcinoma masses appear to be lower than accompanying postobstructive consolidations. ADC values could be considered useful as a differentiating parameter among central lung carcinomas and accompanying postobstructive consolidations. PMID- 19553049 TI - Aging effects on cerebral asymmetry: a voxel-based morphometry and diffusion tensor imaging study. AB - The hemispheres of the human brain are functionally and structurally asymmetric. The purpose of this study was to evaluate the effects of aging on gray and white matter asymmetry. Two hundred twenty-six right-handed normal volunteers aged 21 71 years were included in this study. The effects of aging on gray matter volume asymmetry and white matter fractional anisotropy asymmetry were evaluated with use of voxel-based morphometry and voxel-based analysis of fractional anisotropy maps derived from diffusion tensor imaging (DTI), respectively. The voxel-based morphometry showed no significant correlation between age and gray matter volume asymmetry. The voxel-based analysis of DTI also showed no significant correlation between age and white matter fractional anisotropy asymmetry. Our results showed no significant effects of aging on either gray matter volume asymmetry or white matter fractional anisotropy asymmetry. PMID- 19553051 TI - Measurement of bone cyst fluid volume using k-means clustering. AB - We designed a semiautomatic segmentation method to easily measure the volume of a bone cyst (simple or aneurysmal) from magnetic resonance imaging (MRI). This method only considers the fluid part of the cyst, even when there are several fluid intensities (fluid-fluid levels) or the cyst is multi-loculated. The nonhomogeneity phenomenon inherent in MRI was handled by a k-means clustering algorithm that classified all of the voxels corresponding to the cyst fluid as the same voxel intensity. Level-set segmentation was expanded into the whole cyst volume and the resulting segmented volume provided the measured cyst volume. The semiautomatic method was compared with the usual manual method (manual contour tracing) in terms of its ability to measure a known volume of water (gold standard) as well as the volume of 29 bone cysts. Both methods were equivalent with regards to the gold standard, but the semiautomatic method was more accurate. In terms of the experimental measurements, the semiautomatic method was more repeatable and reproducible, and less time-consuming and fastidious than the manual method. Our semiautomatic method uses only freeware and can be used routinely whenever measurement of a bone cyst volume is needed. PMID- 19553050 TI - MAgnitude and PHase Thresholding (MAPHT) of noisy complex-valued magnetic resonance images. AB - It is often desirable to separate voxels that contain signal from tissue along with measurement noise from those that contain purely measurement noise. Generally, this separation called thresholding utilizes only the magnitude portion of the images. Recently, methods have been developed that utilize both the magnitude and phase for thresholding voxels. This manuscript is an extension previous work and uses the bivariate normality of the real and imaginary values with phase coupled means. A likelihood ratio statistic is derived that simplifies to a more familiar form that is F-distributed in large samples. It is shown that in small samples, critical values from Monte Carlo simulation can be used to threshold this statistic with the proper Type I and Type II error rates. This method is applied to susceptibility weighted magnetic resonance images and shown to produce increased tissue contrast. PMID- 19553053 TI - Viewing the effective k-space coverage of MR images: phantom experiments with fast Fourier transform. AB - The purpose of this experimental study was to evaluate whether the effective k space coverage of MR images can in principle be viewed after multidimensional Fourier transform back to k-space. A water-soaked sponge phantom providing homogeneous k-space pattern was imaged with different standard MR sequences, utilizing elliptic acquisitions, partial-Fourier acquisitions and elliptic filtering as imaging examples. The resulting MR images were Fourier-transformed to the spatial frequency domain (the k-space) to visualize their effective k space coverage. These frequency domain images are named "backtransformed k-space images." For a quantitative assessment, the sponge phantom was imaged with three dimensional partial-Fourier sequences while varying the partial acquisition parameters in slice and phase direction. By linear regression analysis, the k space coverage as expected from the sequence menu parameters was compared to the effective k-space coverage as observed in the backtransformed k-space images. The k-space coverage of elliptic and partial-Fourier acquisitions became visible in the backtransformed k-space images, as well as the effect of elliptic filtering. The expected and the observed k-space coverage showed a highly significant correlation (r=.99, P<.001). In conclusion, the effective k-space coverage of MR images becomes visible when Fourier-transforming MR images of a sponge phantom back to k-space. This method could be used for several purposes including sequence parameter optimization, basic imaging research, and to enhance a visual understanding of k-space, especially in three-dimensional MR imaging. PMID- 19553052 TI - Self-gated Fourier velocity encoding. AB - Self-gating is investigated to improve the velocity resolution of real-time Fourier velocity encoding measurements in the absence of a reliable electrocardiogram waveform (e.g., fetal magnetic resonance or severe arrhythmia). Real-time flow data are acquired using interleaved k-space trajectories which share a common path near the origin of k-space. These common data provide a rapid self-gating signal that can be used to combine the interleaved data. The combined interleaves cover a greater area of k-space than a single real-time acquisition, thereby providing higher velocity resolution for a given aliasing velocity and temporal resolution. For example, this approach provided velocity spectra with a temporal resolution of 19 ms and velocity resolution of 22 cm/s over an 818 cm/s field-of-view. The method was validated experimentally using a computer controlled pulsatile flow apparatus and applied in vivo to measure aortic-valve flow in a healthy volunteer. PMID- 19553054 TI - In vivo structural analysis of articular cartilage using diffusion tensor magnetic resonance imaging. AB - PURPOSE: The articular cartilage is a small tissue with a matrix structure of three layers between which the orientation of collagen fiber differs. A diffusion weighted twice-refocused spin-echo echo-planar imaging (SE-EPI) sequence was optimized for the articular cartilage, and the structure of the three layers of human articular cartilage was imaged in vivo from diffusion tensor images. MATERIALS AND METHODS: The subjects imaged were five specimens of swine femur head after removal of the flesh around the knee joint, five specimens of swine articular cartilage with flesh present and the knee cartilage of five adult male volunteers. Based on diffusion-weighted images in six directions, the mean diffusivity (MD) and the fractional anisotropy (FA) values were calculated. RESULTS: Diffusion tensor images of the articular cartilage were obtained by sequence optimization. The MD and FA value of the specimens (each of five examples) under different conditions were estimated. Although the articular cartilage is a small tissue, the matrix structure of each layer in the articular cartilage was obtained by SE-EPI sequence with GRAPPA. The MD and FA values of swine articular cartilage are different between the synovial fluid and saline. In human articular cartilage, the load of the body weight on the knee had an effect on the FA value of the surface layer of the articular cartilage. CONCLUSION: This method can be used to create images of the articular cartilage structure, not only in vitro but also in vivo. Therefore, it is suggested that this method should support the elucidation of the in vivo structure and function of the knee joint and might be applied to clinical practice. PMID- 19553055 TI - A novel strategy for reducing phospholipids-based matrix effect in LC-ESI-MS bioanalysis by means of HybridSPE. AB - A novel strategy to minimize phospholipids-based matrix effects in bioanalytical LC-MS/MS assays was evaluated. The phospholipids-based matrix effect was investigated with a commercially available electrospray ionization (ESI) source coupled with a triple quadrupole mass spectrometer. A systematic comparison approach of two sample preparation procedures was performed. In particular, the matrix effect on mass spectrometry response in rat and human plasma samples was studied by comparing sample extracts obtained by means of a conventional plasma protein precipitation with acetonitrile and the novel HybridSPE-Precipitation procedure. The HybridSPE dramatically reduced the levels of residual phospholipids in biological samples, leading to significant reduction in matrix effects. This new procedure which combines the simplicity of precipitation with the selectivity of SPE allows to obtain much cleaner extracts than with conventional procedures. The effective targeted removal of phospholipids and proteins in biological plasma samples achieved with the HybridSPE-Precipitation procedure provides significant improvement in bioanalysis and a practical and fast way to ensure the avoidance of phospholipids-based matrix effects. PMID- 19553056 TI - Interactions of tea tannins and condensed tannins with proteins. AB - Binding parameters for the interactions of four types of tannins: tea catechins, grape seed proanthocyanidins, mimosa 5-deoxy proanthocyanidins, and sorghum procyanidins (mDP=17), with gelatin and bovine serum albumin (BSA) have been determined from isothermal titration calorimetry data. Equilibrium binding constants determined for the interaction with gelatin were in the range 10(4) to 10(6) M(-1) and in the order: sorghum procyanidins > grape seed proanthocyanidins > mimosa 5-deoxy proanthocyanidins > tea catechins. Interaction with BSA was generally weaker, with equilibrium binding constants of < or =10(3)M(-1) for grape seed proanthocyanidins, mimosa 5-deoxy proanthocyanidins and tea catechins, and 10(4)M(-1) for the sorghum procyanidins. In all cases the interactions with proteins were exothermic and involved multiple binding sites on the protein. The data are discussed in relation to the structures and the known nutritional effects of the condensed tannins. PMID- 19553057 TI - Determination of total and lactone form of a new camptothecin derivative gimatecan (ST1481) and its metabolite ST1698 in human plasma by high-performance liquid chromatography with fluorimetric detection. AB - A new sensitive high-performance liquid chromatographic (HPLC) method for the determination of gimatecan (ST1481), a new camptothecin derivative, and its metabolite (ST1698) in plasma sample has been developed. The method consisted of on-line column solid phase extraction of analytes from human plasma, chromatographic separation by isocratic elution, then fluorimetric detection. The limits of quantitation were 0.25 ng/mL for both the analytes. The recovery of the extraction procedure was in the range of 62.8-71.1% for all the compounds. Good linearity (R(2)>0.999) was observed within the calibration ranges studied: 0.25 25 ng/mL for both ST1481 and ST1698. Precision was in the range 1.2-4.3%, and accuracy was always lower than 4.7%. Surprisingly, after administration of ST1481 to humans, plasma concentrations found were higher than expected, while metabolite plasma concentrations were negligible. For this reason, a second calibration curve range was validated to quantify ST1481 in human plasma, ranging from 5 to 200 ng/mL. A good accuracy and precision were obtained, confirming the usefulness of the procedure. By using neutral analytical condition the intact lactone form was estimated in plasma samples from a patient. The lactone form amounted to 80-100% of the total ST1481. PMID- 19553058 TI - Exploring perceptions of genetic testing: an examination of perceived accuracy over time. AB - OBJECTIVE: Despite the potential impact of perceptions of genetic testing, little research has examined these perceptions. The current study examined factors associated with perceived accuracy of genetic testing to detect BRCA1/2 mutations and change in perceived accuracy. METHODS: Participants were recruited through advertisements, community groups, and local oncologists and gynecologists. Ashkenazi Jewish patients (N=120) at increased risk for BRCA1/2 genetic mutations were surveyed in a repeated-measures design with assessments at three time points: pre-counseling, post-counseling, and post-testing. RESULTS: Better knowledge of cancer genetics related to higher perceived accuracy before counseling. Lower perceived risk of developing ovarian cancer was associated with increased perceived accuracy, while greater distress regarding gene status was associated with decreased perceived accuracy. Genetic test results were unrelated to change in perceived accuracy. CONCLUSION: The results offer an initial look at how patients form perceptions of genetic testing. Both cognitive factors and affective factors relate to how patients perceived the accuracy of testing and how their perception changes over time. PRACTICE IMPLICATIONS: Understanding cognitive and affective factors which may impact patients' perceptions of genetic testing is important. Specifically, attention should be paid to educating patients regarding cancer genetics and encouraging adaptive coping strategies to deal with distress. PMID- 19553059 TI - Patient attitudes to being treated by junior residents in the community. AB - OBJECTIVE: Understanding the attitudes of patients to being treated by residents in the community. METHODS: A questionnaire was administered to patients visiting community family medicine teaching clinics. The study methodology included statements to which they agree or disagree. RESULTS: Three hundred and four questionnaires were completed by patients; 94% had visited a resident in the past year; 78.9% agreed that residents were as skilled as senior doctors, but only 45.4% felt that they were as quick at diagnosis as the senior doctors; 73.0% felt that residents spent more time with them; 40.0% were not pleased by the constant change of the residents attending on them. Analysis by logistic regression showed that men had a more positive attitude to the competence of the residents as well as their professionalism (OR 2.73, 95% CI, 1.45-5.10). Frequent visitors to the clinic had a more negative attitude to the residents' professionalism (OR 0.91 (0.85-0.98)) and were more likely to agree with the statement "I would prefer to see the regular doctor and not a different resident each time" (OR 1.09 (1.01 1.18)). Those who were attended more by residents on their visits showed a positive attitude to the professionalism of residents (OR 1.14 (1.01-1.28)) and were less likely to agree with the statement "I would prefer to see the regular doctor and not a different resident each time" (OR 0.90 (0.84-0.98)). CONCLUSIONS: Patients have a positive attitude to being treated by residents in ambulatory clinics, which is associated with repeat exposure to care by residents. PRACTICAL IMPLICATIONS: Departments with ambulatory training should consider having constant presence of residents in their teaching clinics, and teaching staff in the clinics should develop ways to recommend patients to be seen by residents. PMID- 19553060 TI - Studying empathy as an interactional three-part sequence. PMID- 19553061 TI - Buprenorphine maintenance treatment in a primary care setting: outcomes at 1 year. AB - The purposes of this study were to assess outcomes of patients prescribed buprenorphine at a primary care practice and to identify factors associated with favorable outcomes. All 255 patients given at least one prescription for buprenorphine between August 2003 and September 1, 2007, at a primary care practice in Baltimore were included. Data regarding demographics and comorbidities were collected retrospectively. Patients were classified as "opioid positive" or "opioid-negative" each month based on patient report, urine toxicology, and provider assessment. After 12 months, 145 (56.9%) patients remained in treatment, and 64.7% of their months were opioid-negative. Patients using heroin were less likely to be opioid-negative, whereas those using prescription opioids were more likely to be opioid-negative. Polysubstance use was associated with increased treatment retention. The prescription of buprenorphine for opioid dependence treatment can be incorporated into primary care practice, and many patients, including polysubstance users, benefit from this treatment. PMID- 19553062 TI - Choosing a control group in effectiveness trials of behavioral drug abuse treatments. AB - Effectiveness trials are an important step in the scientific process of developing and evaluating behavioral treatments. The focus on effectiveness research presents a different set of requirements on the research design when compared with efficacy studies. The choice of a control condition has many implications for a clinical trial's internal and external validity. The purpose of this article was to provide a discussion of the issues involved in choosing a control group for effectiveness trials of behavioral interventions in substance abuse treatment. The authors provide a description of four trial designs and a discussion of the advantages and disadvantages of each. PMID- 19553063 TI - Behavioral couple therapy for gay and lesbian couples with alcohol use disorders. AB - Gay (n = 52) and lesbian (n = 48) patients with alcohol use disorder (AUD) and their non-substance-abusing same-sex relationship partners were randomly assigned to equally intensive interventions consisting of (a) behavioral couples therapy (BCT) plus individual-based treatment (IBT) or (b) IBT only. This study reports two separate trials, one with gay male participants and one with lesbian female participants. For both gay and lesbian patients with AUD, those who received BCT had a significantly lower percentage of days of heavy drinking during the year after treatment than patients who received IBT only. In addition, both gay and lesbian couples who received BCT reported higher levels of relationship adjustment at the end of treatment and in the year after treatment than those who received IBT only. Thus, the response of gay and lesbian couples with an alcoholic member to BCT was consistent with what has been observed with heterosexual couples. PMID- 19553065 TI - Reducing cultural barriers to substance abuse treatment among Asian Americans: a case study in New York City. AB - This article addresses the issue of underutilization of treatment services by Asian Americans. Although Asian Americans continue to be the fastest growing ethnic minority in New York City, they are reported to underutilize substance abuse treatment services. However, much of the underutilization may be attributed to cultural and language barriers hindering them from seeking help and support. Past literature indicates that when culturally appropriate services are available, service utilization tends to increase among Asian Americans. This study tested strategies to enhance the continuum of care in the Asian community by adapting a well-documented generic early intervention model in a culture specific setting. The results of this study indicate that these models may be successfully adapted to culturally specific settings such as Asian American communities. When culturally competent services combined with case management and motivational interviewing are provided, there tends to be an increase in Asian clients' chance of accomplishing treatment goals. PMID- 19553064 TI - College students rarely seek help despite serious substance use problems. AB - The prevalence of substance use disorders (SUD) and aspects of the help-seeking process among a high-risk sample of 946 students at one large public university were assessed in personal interviews during the first 3 years of college. After statistically adjusting for purposive sampling, an estimated 46.8%(wt) of all third-year students met Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for SUD involving alcohol and/or marijuana at least once. Of 548 SUD cases, 3.6% perceived a need for help with substance use problems; 16.4% were encouraged by someone else to seek help. Help-seeking was rare among SUD cases (8.8%) but significantly elevated among individuals who perceived a need (90.0%) or experienced social pressures from parents (32.5%), friends (34.2%), or another person (58.3%). Resources accessed for help included educational programs (37.8%), health professionals (27.0%), and 12-step programs (18.9%). College students have high rates of substance use problems but rarely recognize a need for treatment or seek help. Results highlight the opportunity for early intervention with college students with SUD. PMID- 19553066 TI - Health-related quality of life trajectories of methamphetamine-dependent individuals as a function of treatment completion and continued care over a 1 year period. AB - This study applies a chronic illness framework to evaluate treatment outcomes among individuals dependent on methamphetamine (MA). Using growth curve modeling, health-related quality of life (HRQOL) trajectories of MA-dependent individuals (N = 723) were examined over a 1-year period. Results show greater improvements in mental HRQOL trajectories as a function of treatment completion and continued care, although fairly static trajectories in physical health status. Other factors affecting HRQOL trajectories included gender, psychosocial functioning, drug use severity, and health impairment. Results extend research on treatment evaluations for MA dependence, highlighting the importance of continued service utilization for improved quality of life outcomes. PMID- 19553068 TI - JNK/AP-1 pathway is involved in tumor necrosis factor-alpha induced expression of vascular endothelial growth factor in MCF7 cells. AB - Vascular endothelial growth factor (VEGF) has been implicated in breast tumor angiogenesis. And tumor necrosis factor-alpha (TNF-alpha) is a positive regulator of VEGF. This study was aimed to identify the signalling pathway of TNF-alpha in VEGF expression regulation in breast cancer cell line MCF7. Using luciferase reporter assays, we demonstrated that TNF-alpha significantly increased activator protein-1 (AP-1) transcriptional activity in the MCF7 cells. The expression of the AP-1 family members c-Jun, c-Fos and JunB and phosphorylation levels of c-Jun were upregulated by TNF-alpha, whereas other AP-1 family members Fra-1, Fra-2, and JunD were unaffected. The activation of AP-1 was associated with the formation of p-c-Jun-c-Jun and p-c-Jun-JunB homodimers. Furthermore, the phosphorylation levels of c-Jun N-terminal kinase (JNK) but not P38 and ERK were elevated by TNF-alpha in MCF7 cells. TNF-alpha potently upregulated the mRNA and protein levels of VEGF, which were significantly reversed by JNK inhibitor SP600125. Finally using chromatin immunoprecipitation (CHIP) assays, we found that p-c-Jun bound to the VEGF promoter and regulated VEGF transcription directly. These data suggest that the pro-inflammatory cytokine TNF-alpha is a critical regulator of VEGF expression in breast cancer cells, at least partially via a JNK and AP-1 dependent pathway. PMID- 19553067 TI - The Washington Circle continuity of care performance measure: predictive validity with adolescents discharged from residential treatment. AB - This study examined the predictive validity of the Washington Circle (WC) continuity of care after long-term residential treatment performance measure, as well as the impact of assertive continuing care interventions on achieving continuity of care. This measure is a process measure that focuses on timely delivery of a minimal floor of services that are necessary to provide sufficient quality of treatment but should not be construed to be the optimal continuity of care after residential treatment for any specific adolescent. Participants included 342 adolescents who were admitted to long-term residential treatment and randomly assigned to either standard continuing care or an assertive continuing care condition. Overall, results provide initial support for the WC continuity of care after residential treatment performance measure as a useful predictor of 3 month recovery status. In addition, assignment to an assertive continuing care condition was found to significantly increase the likelihood of achieving continuity of care. PMID- 19553069 TI - Can an anterior quadriceps release improve range of motion in the stiff arthritic knee? AB - We hypothesize that tethering adhesions of the quadriceps muscle are the major pathological structures responsible for a limited range of motion in the stiff arthritic knee. Forty-two modified quadriceps muscle releases were performed on 24 patients with advanced osteoarthritis scheduled for total knee arthroplasty. The ranges of motion were documented intraoperatively both before and immediately after the release. Passive flexion improved significantly in all patients (mean, 32.4 degrees of improvement, P < .001) following a modified quadriceps release, despite any presence of osteophytes or severe deformities. These results strongly implicate adhesions of the quadriceps muscle to the underlying femur, which prevent the distal excursion of the quadriceps tendon, as the restrictive pathology preventing deep flexion in patients with osteoarthritis. PMID- 19553070 TI - Clinical experience of Ganz surgical dislocation approach for metal-on-metal hip resurfacing. AB - Although the posterior approach is the most commonly used for hip resurfacing, concerns remain in terms of risk of femoral neck fracture secondary to an osteonecrotic event. The purpose of this study was to look at the short-term results of metal-on-metal hip resurfacing done by the vascular-preserving surgical approach as developed by Ganz in 116 hip resurfacing arthroplasties performed in 106 patients (86 men, 20 women; mean age, 46.5 years; range, 19-62). At a mean follow-up of 38.3 months (range, 12-84), Harris Hip Scores improved significantly from 53.1 to 90.16 (P < .001). There were 10 nonunions (8.7%) and 21 hips (18.3%) requiring screw removal for painful bursitis. Two hips underwent conversion to total hip arthroplasty: one at 18 months for femoral loosening and one at 7 years for acetabular loosening. Although the trochanteric slide approach as developed by Ganz provides excellent exposure to the hip joint and preserves femoral head vascularity, it does carry some inherent morbidity in regard to the greater trochanter. PMID- 19553071 TI - Major complications associated with femoral nerve catheters for knee arthroplasty: a word of caution. AB - One thousand one hundred ninety patients underwent knee arthroplasty between January 2004 and July 1, 2007, and received an indwelling continuous infusion femoral catheter for postoperative ropivicaine pump infusion. Catheters were placed using electrical stimulation guidance. For the initial 469 patients (group 1), the continuous infusion ran for 2 to 3 days. In 721 patients, the infusion was discontinued 12 hours after surgery. There were 9 femoral nerve palsies (2 in group 1, 7 in group 2) and 8 major falls (0.7%). The overall complication rate was 1.5%, and the risk of permanent nerve injury was 0.2%. Patients should be made aware of these complications as part of the usual informed consent process before using this technique for postoperative pain control after knee arthroplasty. We did not observe fewer falls when the continuous infusion was stopped 12 hours after surgery. PMID- 19553072 TI - Fixed- vs mobile-bearing total knee arthroplasty: does it make a difference?--a prospective randomized study. AB - The purpose of this prospective, randomized study was to compare the early clinical and functional results of primary total knee arthroplasty using a fixed bearing (FB) and a rotating-platform (RP) prosthesis. Outcomes including range of motion (ROM), Knee Society Score, Western Ontario MacMaster (WOMAC), and Short Form-36 (SF-36) were measured preoperatively and at 6 weeks, 3 months, 6 months, 1 year, and 2 years. Radiographic analysis was performed. There were 72 FB and 68 RP knees. The RP group had a greater ROM at 6 weeks and 1 year. This difference was not statistically significant at 2 years. There were no differences in the ROM at any other period. There were no significant differences in Knee Society Score, Short Form-36, or Western Ontario MacMaster scores at any period. No clinically significant differences were noted in the radiographic analysis. The use of a FB or RP design did not affect the early functional outcomes after total knee arthroplasty. PMID- 19553073 TI - Coronal alignment in total knee arthroplasty: just how important is it? AB - A recent study has challenged the premise that well-aligned total knee arthroplasties (TKAs) have better survival than outliers. This study examines the importance of overall coronal alignment as a predictor for revision. Patients with primary TKAs were stratified into neutral, varus, and valgus alignment groups based on the postoperative tibiofemoral angle. In 6070 knees (3992 patients), there were 51 failures (0.84%): 21 (0.5%) in the neutral group, 18 (1.8%) in the varus group, and 12 (1.5%) in the valgus group. The best survival was for overall alignment between 2.4 degrees and 7.2 degrees of valgus. Varus knees failed primarily by medial tibia collapse, whereas valgus knees failed from ligament instability. Outliers in overall alignment have a higher rate of revision than well-aligned knees. The goal of TKA should be to restore alignment within 2.4 degrees to 7.2 degrees of valgus. PMID- 19553074 TI - Long-term results of cemented total hip arthroplasty for dysplasia, with structural autograft fixed with poly-L-lactic acid screws. AB - This study reviewed a series of cemented total hip arthroplasty (THA) for dysplasia, with structural autograft fixed with poly-L-lactic acid screws. Grafted bone union was confirmed radiologically in every case, and there were no cases of early collapse or extravasation of grafted bone. Kaplan-Meier survivorship analysis of socket revision, radiologic loosening of the socket, and the appearance of a radiolucent line greater than 1 mm in the graft-socket interface as the end points indicated survival rates of 99%, 97.1%, and 63.5% at 10 years and 96.6%, 90.2%, and 56.1% at 15 years, respectively. The results of this study indicated that poly-L-lactic acid screws are safe and useful for the fixation of acetabular bone graft concomitant to cemented THA with a careful rehabilitation program. PMID- 19553075 TI - A validation model for measurement of acetabular component position. AB - There is no agreement on a standard approach to evaluating acetabular cup orientation, ideal target orientation, or a standardized measurement method for cup orientation in total hip arthroplasty. The purpose of this study was to investigate a simple method for validating measurements of acetabular orientation obtained using computer navigation and computed tomography scans. This study validated the imageless navigation system to be accurate with a precision of 1 degrees and a bias of 0.02 degrees for inclination and a precision of 1.3 degrees and a bias of 0 degrees for anteversion measurements. From this study, we propose that acetabular cup alignment is accurately assessed using computer navigation. We suggest acetabular orientation be reported in the radiographic plane (coronal plane), which incorporates pelvic tilt and therefore is more functional definition of cup position. PMID- 19553076 TI - Clinical and radiographic results of 184 consecutive revision total knee arthroplasties placed with modular cementless stems. AB - Clinical and radiographic outcomes of 184 consecutive revision total knee arthroplasties (TKAs) placed with cemented components and press-fit fluted cementless stems were reviewed at average follow-up of 49 months. Revision was performed for septic failure in 53 of 184 knees, periprosthetic fracture in 10 of 184 knees, and aseptic failure in 121 of 184 knees. The press-fit cementless stems were placed to tightly contact the endosteum of the metadiaphyseal area of the femur and tibia. No knees were re-revised for aseptic loosening. There were 15 failures (15/184, 8.2%): 13 were septic failures, 9 of which were recurrence of sepsis and 4 of which were new cases of sepsis. Average Knee Society knee score improved from 134.6 to 167.8 (P < .05). Revision TKA with modular cementless stems provided excellent fixation with no revisions for aseptic loosening. Re-revision for sepsis was the dominant failure mechanism with an overall rate of 7%. Given the increasing prevalence of revision TKA, sepsis prevention strategies should receive increased attention. PMID- 19553077 TI - The fate of acute methicillin-resistant Staphylococcus aureus periprosthetic knee infections treated by open debridement and retention of components. AB - The success of open irrigation and debridement with component retention (ODCR) for acute periprosthetic knee joint infection varies widely. The species and virulence of the infecting organism have been shown to influence outcome. This multicenter, retrospective study identified 19 cases of acute periprosthetic methicillin-resistant Staphylococcus aureus (MRSA) knee infections managed by ODCR and at least 4 weeks of postoperative intravenous vancomycin therapy. At minimum follow-up of 2 years, the treatment failed to eradicate the infection in 16 cases (84% failure rate). Of those 16 failures, 13 patients required a 2-stage exchange arthroplasty, 2 patients required repeat incision and debridement with antibiotic suppression, and 1 patient died of MRSA sepsis. In addition, a systematic review of the literature revealed failure to eradicate infection in 10 of 13 patients managed with a similar protocol. The total success rate of ODCR in acute periprosthetic MRSA knee infection was 18%. PMID- 19553078 TI - Social fears during adolescence: is there an increase in distress and avoidance? AB - Mid-adolescence is considered as the time of onset for social phobia and is assumed to be related to a normative increase of social fears. People diagnosed with social phobia, however, do not only experience high levels of fear or distress, but also report avoidance behavior. Little attention has been paid to the development of avoidance behavior during adolescence. In the current study, a community sample with 9-17 year olds (n=260) completed a questionnaire derived from the Anxiety Disorders Interview Schedule for Children (ADIS-C) [Silverman, W. K., & Albano, A. M. (1996). Anxiety disorders interview schedule for DSM-IV child version, child interview schedule. San Antonio: The Psychological Corporation]. They rated their levels of distress and avoidance in a variety of social situations. The results showed an age related increase for formal speaking and interaction situations in both avoidance and distress, with a stronger increase in avoidance than in distress. The same pattern was found for girls for situations regarding observation by others. No effects were observed for informal speaking and interaction situations. PMID- 19553079 TI - Assessment of left ventricular relaxation by untwisting rate based on different algorithms. AB - BACKGROUND: This study was performed in both animals and human subjects to test whether different approaches to calculate untwisting rate may lead to different results in the assessment of left ventricular (LV) relaxation. METHODS: In animal experiments, congestive heart failure was successfully induced in 8 adult dogs. Transthoracic echocardiography was performed with simultaneous LV pressure recording at baseline and the stage of heart failure. In the clinical study, 72 patients undergoing right-sided heart catheterization were studied by transthoracic echocardiography. LV twist was calculated as the difference between apical and basal rotations measured using two-dimensional speckle tracking. Untwisting rate was calculated using 3 different algorithms as the peak negative time derivative of twist (UR(max)) during early diastole, the slope of the linear regression of untwisting over time (UR(slope)), or the average untwisting over the isovolumic relaxation period (UR(mean)). RESULTS: UR(max) significantly correlated with tau and -dP/dt in dogs (r=-0.81 and 0.77, respectively, both P < .001) and was reduced at the stage of heart failure (P < .01). In 55 patients (76%) with adequate image quality, only UR(max) among untwisting rates calculated by 3 different algorithms was significantly related to tau (r=-0.51, P < .001). UR(max) was significantly lower in patients with tau>/=48 ms than in patients with tau<48 ms (P=.004), most of whom had a depressed LV ejection fraction. CONCLUSION: UR(max) best reflects LV relaxation in comparison with the 2 other algorithms. PMID- 19553080 TI - Echocardiographic tissue deformation imaging quantifies abnormal regional right ventricular function in arrhythmogenic right ventricular dysplasia/cardiomyopathy. AB - BACKGROUND: The aim of this study was to determine the accuracy of new quantitative echocardiographic strain and strain-rate imaging parameters to identify abnormal regional right ventricular (RV) deformation associated with arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C). METHODS: A total of 34 patients with ARVD/C (confirmed by Task Force criteria) and 34 healthy controls were prospectively enrolled. Conventional echocardiography, including Doppler tissue imaging (DTI), was performed. Doppler and two dimensional strain-derived velocity, strain, and strain rate were calculated in the apical, mid, and basal segments of the RV free wall. RESULTS: RV dimensions were significantly increased in patients with ARVD/C (RV outflow tract 19.3+/-5.2 mm/m2 vs 14.1+/-2.2 mm/m2, P<.001; RV inflow tract 23.4+/-4.8 mm/m2 vs 18.8+/-2.4 mm/m2, P<.001), whereas left ventricular dimensions were not significantly different compared with controls. Strain and strain rate values were significantly lower in patients with ARVD/C in all 3 segments. All deformation parameters showed a higher accuracy to detect functional abnormalities compared with conventional echocardiographic criteria of dimensions or global systolic function. The lowest DTI strain value in any of the 3 analyzed segments showed the best receiver operating characteristics (area under the curve 0.97) with an optimal cutoff value of -18.2%. CONCLUSIONS: DTI and two-dimensional strain derived parameters are superior to conventional echocardiographic parameters in identifying ARVD/C. This novel technique may have additional value in the diagnostic workup of patients with suspected ARVD/C. PMID- 19553081 TI - Doppler assessment of the ratio of the systolic to diastolic duration in normal children: relation to heart rate, age and body surface area. AB - BACKGROUND: It has been shown that the ratio of systolic duration to diastolic duration (S/D) is a valuable global index of ventricular dysfunction in pediatric dilated and restrictive cardiomyopathy and is also a valuable index of the ventricular function of single systemic right ventricles in children who have undergone palliation of hypoplastic left-heart syndrome. The purposes of this study were to establish normal values for the S/D ratio in children without heart disease and to investigate its variation with heart rate (HR), age, and body surface area. METHODS: Of 752 children found to have functional murmurs by clinical means, 179 (24% of the total database) had trace holosystolic tricuspid regurgitation. These patients ranged in age from 0.02 months to 19 years. For this study, the diastolic interval was defined as the period of right ventricular filling (ie, the period between 2 tricuspid regurgitant jets) and the systolic interval as the remainder of the cardiac cycle corresponding to the duration of holosystolic tricuspid regurgitation, including the periods of isovolumic contraction and relaxation. The relations between the systolic and diastolic periods and their ratio (S/D ratio) and HR, age, and body surface area were evaluated using univariate and multivariate linear regression analysis. RESULTS: Patient's age ranged from 0.02 months to 19 years (mean, 70.18+/-65.12 months), body surface area from 0.11 to 2.51 m2 (mean, 0.85+/-0.55 m2), and HR from 50 to 156 beats/min (mean, 96.72+/-23.19 beats/min). The systolic period ranged from 208.5 to 467 ms (mean, 314.08+/-52.57 ms) and the diastolic period from 166.5 to 809 ms (mean, 341.34+/-129.61 ms), yielding a S/D ratio ranging from 0.397 to 1.62 (mean, 0.995+/-0.23). Systolic period duration showed a linear negative decrease with increasing HR (y=1.9228x+500.05, r=-0.85). Diastolic duration decreased in an exponential fashion as HR increased (y=130,679x(-1.3232), r= 0.88). The S/D ratio correlated positively with HR (y=-1.656+0.0265 HR-0.000954 HR2, r=76). On multivariate analysis, no significant correlations of S/D ratio with age or body surface area were found. CONCLUSIONS: The results of this study provide useful reference values for the S/D ratio across a wide range of HRs in children, adolescents, and young adults. PMID- 19553082 TI - Multiplanar reconstruction of three-dimensional transthoracic echocardiography improves the presurgical assessment of mitral prolapse. AB - BACKGROUND: The aim of this study was to evaluate the value and accuracy of multiplanar reconstruction (MPR) of three-dimensional (3D) transthoracic echocardiographic data sets in assessing mitral valve pathology in patients with surgical mitral valve prolapse (MVP). METHODS: Sixty-four patients with surgical MVP and preoperative two-dimensional (2D) and 3D transthoracic echocardiography were analyzed. The descriptions obtained by 3D MPR and 2D were compared in the context of the surgical findings. RESULTS: Two-dimensional echocardiography correctly identified the prolapsing leaflets in 32 of 64 patients and 3D MPR in 46 of 64 patients (P=.016). Among the 27 patients with complex pathology (ie, more than isolated middle scallop of the posterior leaflet prolapse), 3D MPR identified 20 correctly, as opposed to 6 with 2D imaging (P<.001). CONCLUSION: Interpretation of 3D transthoracic echocardiographic images with MPR improved the accuracy of the description of the MVP compared with 2D interpretation. This added value of 3D MPR was most important in extensive and/or commissural prolapse. PMID- 19553083 TI - Left atrial systolic force in children: reference values for normal children and changes in cardiovascular disease with left ventricular volume overload or pressure overload. AB - BACKGROUND: Recent studies in adults have indicated that left atrial (LA) systolic force (LASF) provides useful information about LA pump function and can be used to predict cardiovascular events. However, normal values of LASF in children are not available, and little is known about atrial function in pediatric patients with heart disease. The objectives of the present study were to provide reference values for LASF in children and to determine LA pump performance in pediatric patients with heart disease using LASF. METHODS: LASF was measured using combined two-dimensional imaging and Doppler echocardiography in 185 healthy children and 71 pediatric patients with ventricular septal defects (VSDs; as a representative heart disease with chronic left ventricular [LV] volume overload; n=48) or coarctation of the aorta (COA; as a representative heart disease with chronic LV pressure overload; n=23). RESULTS: LASF in children significantly increased with advancing age (P<.005). The major determinants of this change were body surface area, stroke volume, and heart rate, with a linear model fit (r2) of 0.72. In patients with VSD and those with COA, LASF was significantly elevated in proportion to the degree of LV volume or pressure load, suggesting adaptive mechanisms of LA pump function to facilitate LV filling in chronically overloaded hearts. LASF was normalized after anatomic correction of VSDs but remained elevated even after the relief of COA, indicating persistent overwork of the left atrium. CONCLUSIONS: The present study provides reference data for the evaluation of atrial function in pediatric patients with cardiac disease. In addition, altered LASF in patients with VSDs with even small left-to right shunts and in those with COA even after anatomic correction may have pathologic implications that could lead to a novel therapeutic target. PMID- 19553084 TI - Prognostic value of dipyridamole stress myocardial contrast echocardiography: comparison with single photon emission computed tomography. AB - BACKGROUND: Dipyridamole stress myocardial contrast echocardiography (MCE) can be used to detect coronary artery disease (CAD). Because it measures myocardial blood flow velocity in addition to measuring myocardial blood volume, it was hypothesized that it should have greater prognostic utility than single photon emission computed tomography (SPECT), which measures only myocardial blood volume. Because blood flow mismatch precedes wall thickening (WT) abnormalities during demand ischemia, it was also postulated that perfusion on MCE would be superior to WT abnormalities on echocardiography for this purpose. METHODS: The incidence of nonfatal myocardial infarction and cardiac death was determined in 261 patients with known or suspected CAD over a mean follow-up period of 14 months who underwent simultaneous dipyridamole stress MCE and 99mTc-sestamibi SPECT. Comparisons of survival curves were conducted with stratified (and unstratified) log-rank tests. RESULTS: Abnormal results on MCE were found to be the best predictor of an adverse outcome (odds ratio, 23; 95% confidence interval, 6-201; P<.0001) and provided incremental prognostic value over clinical variables (age>60 years, the presence of >or=3 cardiac risk factors, known peripheral vascular disease, prior myocardial infarction, and left ventricular systolic function), inducible WT abnormalities, and SPECT. Prognoses were worst in patients who had both abnormal results on MCE and inducible WT abnormalities and best in those who had neither. Patients with abnormal results on MCE but no inducible WT abnormalities had intermediate outcomes. CONCLUSION: In patients with known or suspected CAD undergoing dipyridamole stress, MCE provides powerful prognostic information that is superior to clinical variables, electrocardiography, left ventricular systolic function, WT analysis, and SPECT. MCE may therefore serve as a method of choice for myocardial perfusion assessment in patients with known or suspected CAD. Larger studies are needed to confirm these findings. PMID- 19553085 TI - Role of serotoninergic pathways in drug-induced valvular heart disease and diagnostic features by echocardiography. AB - Serotonin plays a significant role in the development of carcinoid heart disease, which primarily leads to fibrosis and contraction of right-sided heart valves. Recently, strong evidence has emerged that the use of specific drug classes, such as ergot alkaloids (for migraine headaches), 5-hydroxytryptamine (5-HT or serotonin) uptake regulators or inhibitors (for weight reduction), and ergot derived dopamine agonists (for Parkinson's disease), can result in left-sided heart valve damage that resembles carcinoid heart disease. Recent studies have suggested that both right-sided and left-sided drug-induced heart valve disease involves increased serotoninergic activity and in particular activation of the 5 HT receptors, including the 5-HT2B receptor subtype, which mediate many of the central and peripheral functions of serotonin. G-proteins that inhibit adenylate cyclase activity mediate the activity of the 5-HT2B receptor subunit, which is widely expressed in a variety of tissues, including liver, lung, heart, and coronary and pulmonary arteries; it has also been reported in embryonic mouse heart, particularly on mouse heart valve leaflets. In this review, the authors discuss the salient features of serotoninergic manifestations of both carcinoid heart disease and drug-induced cardiac valvulopathy, with an emphasis on echocardiographic diagnosis. PMID- 19553086 TI - Effectiveness of second-generation antipsychotics with acute-phase schizophrenia. AB - PURPOSE: Although olanzapine may have advantages over other second-generation antipsychotics (SGAs) regarding longer time to treatment discontinuation among chronically ill patients, little evidence has been provided for the comparative effectiveness of SGAs in the acute phase. We aimed to determine if any of four SGAs were more effective in treating newly admitted acute schizophrenic patients. We performed a rater-blinded, randomized controlled trial of four SGAs in 15 psychiatric emergency sites. Eligible patients were 18-64 years old and met diagnostic criteria for schizophrenia, acute schizophrenia-like psychotic disorder, or schizoaffective disorder. A final total of 78 patients were randomly assigned by means of sealed envelopes to receive risperidone (3-12 mg/day; n=20), olanzapine (10-20 mg/day; n=17), quetiapine (300-750 mg/day; n=20), or aripiprazole (12-30 mg/day; n=21), with follow-up at 8 weeks. The primary outcome measure was all-cause treatment discontinuation. RESULTS: Overall, 37% (29/78) of patients discontinued the study medication before 8 weeks: 25% for risperidone; 12% for olanzapine; 55% for quetiapine; and 52% for aripiprazole. Time to treatment discontinuation for any cause was significantly longer in the olanzapine group than in the quetiapine (p=0.006) or aripiprazole (p=0.008) groups, but not compared to the risperidone group (p=0.32). Time to treatment discontinuation was significantly longer in the risperidone group than in the quetiapine group (p=0.048), but not compared to the aripiprazole group (p=0.062). However, the rate of p.r.n. intramuscular haloperidol use was significantly higher in the aripiprazole group than in other groups (p=0.029). CONCLUSION: Olanzapine and risperidone are superior to quetiapine and aripiprazole for the acute treatment of psychosis in hospitalized patients. PMID- 19553087 TI - Deficit schizophrenia: an overview of clinical, biological and treatment aspects. AB - The concept of deficit schizophrenia is regarded as one of the most promising attempts to reduce heterogeneity within schizophrenia. This paper summarizes the clinical, neurocognitive, brain imaging and electrophysiological correlates of this subtype of schizophrenia. Attempts to identify genetic and non-genetic risk factors are reviewed. Methodological limitations of studies supporting the efficacy of atypical antipsychotics in the treatment of the syndrome are highlighted. Two decades of research on deficit schizophrenia have failed to prove that it represents the extreme end of a severity continuum in schizophrenia, while some findings support the claim that it may be a separate disease entity. PMID- 19553088 TI - Changes in lifestyle for psychiatric patients three years after the start of short- and long-term psychodynamic psychotherapy and solution-focused therapy. AB - OBJECTIVES: Lifestyle is less favourable among individuals suffering from psychiatric disorders. We studied whether psychotherapy brings along changes in lifestyle and whether these changes differ between short-term and long-term psychodynamic psychotherapy (SPP and LPP) and solution-focused therapy (SFT). METHODS: A total of 326 outpatients, 20-46 years of age, with mood or anxiety disorder were randomly assigned to LPP, SPP and SFT. The lifestyle variables considered were alcohol consumption, smoking, body mass index (BMI), leisure time exercise and serum cholesterol. The patients were monitored for three years from the start of treatment. RESULTS: During the three-year follow-up, BMI and serum cholesterol rose statistically significantly although no statistically significant trends were shown for alcohol consumption, smoking or exercise. SPP showed a disadvantage of increased alcohol consumption and serum cholesterol level when compared with LPP. SFT showed an advantage of reduced smoking in comparison with SPP. DISCUSSION: Small therapy-specific changes in lifestyle may be a result from psychotherapy treatment. These lifestyle changes are apparently more common in short-term therapy. More studies are needed to verify these findings. PMID- 19553090 TI - Evaluation of psychiatric liaison-services for adolescents in residential group homes. AB - OBJECTIVE: Adolescents in residential care are a high-risk population for psychiatric disorders. Due to limited access to outpatient treatment, crises often result in psychiatric hospitalizations. These crises are often accompanied by termination of supportive relationships, which increases the risk for further problems. To minimize hospitalizations and to stabilize the psychological state of these adolescents, we developed and evaluated a home-treatment intervention program based on psychiatric liaison-services and compared it to standard medical care (SMC). METHODS: The intervention was applied in 11 German residential group homes (N = 288, M = 14.1 years, SD = 2.7), while adolescents from 15 institutions (N = 336, M = 13.6 years, SD = 3.0) served as a comparison group (CG) (SMC). The frequency and duration of psychiatric inpatient treatment episodes were documented and caregiver-reported behavioural symptoms were assessed at baseline and at 6 and 12 months postintervention. RESULTS: In both groups, less than 10% of the adolescents were admitted to psychiatric wards. Compared to SMC, hospitalizations were of shorter duration in the intervention group (IG) (1.30 vs 2.47 days per head within 12 months, p < .001). In both groups, behavioural problems decreased overtime. CONCLUSIONS: Applying a home-treatment intervention program to children in residential care helps to stabilize the developmental course of this high-risk population by shortening psychiatric inpatient treatment. PMID- 19553089 TI - Norwegian version of the Mini-International Neuropsychiatric Interview: feasibility, acceptability and test-retest reliability in an acute psychiatric ward. AB - OBJECTIVE: To assess the feasibility, patient and clinician acceptability and test-retest reliability of the Mini-International Neuropsychiatric Interview (MINI) used by non-psychiatrists in an acute psychiatric ward. METHOD: Of 268 consecutive patients included in a cross-sectional study, 176 (66%) completed MINI, and were compared to patients not interviewed. Patients and clinicians were questioned about the interview, using Visual Analogue Scales (VAS). For 38 patients, test-retest reliability was assessed with Cohen's kappa and observed agreement. RESULTS: MINI was not feasible for all patients. Among factors associated with not being interviewed were early discharge, psychosis, substance use and involuntary admissions. Although evaluations by patients and clinicians completing the postinterview questionnaire varied, MINI was generally perceived as being useful and feasible. Psychotic symptoms were associated with a less positive experience with MINI for both patients and clinicians. In the test retest analyses, kappa values indicated excellent agreement for six diagnoses, fair to good for six and poor for seven, whereas observed agreement was 75% or above for all disorders. CONCLUSION: Among patients admitted to an acute psychiatric ward willing and able to complete the interview, MINI was well accepted by patients and clinicians, and has moderately good test-retest reliability. PMID- 19553091 TI - Development of the self-administered Spanish version of the Health of the Nation Outcome Scales for Older Adults: HoNOS65+A. AB - OBJECTIVES: The purpose of this study is to develop a self-administered version of the Health of the Nation Outcome Scales for Older Adults (HoNOS65+) in Spanish - the HoNOS65+A - and to study its reliability and validity. METHODS: There are two phases in the study: construction of the HoNOS65+A and the study of its psychometric properties. In the second phase, 179 users of 10 services for older people of Madrid were interviewed. They completed the HoNOS65+A, the Spanish version of the HoNOS65+, and other instruments that evaluate similar constructs. Reliability (internal consistency with the other-administered HoNOS65+, intraclass) and validity (concurrent, discriminant, and criterion validity) were analyzed. RESULTS: High reliability indexes (intraclass correlation coefficients) were found for all the scales (HoNOS65+ and HoNOS65+A), except for items 4 and 6. Moreover, the HoNOS65+A has satisfactory concurrent (except 4, 6, and 9) and discriminant validity (except for items 4 and 10). Considering all these data and some theoretical points the items 4 and 6 have eliminated from the scale. Upon eliminating items 4 and 6, the analysis of the reliability (Cronbach's Alpha) and criterion validity (discriminant function among autonomous and assisted people) presents positive values. CONCLUSION: The results indicate that the HoNOS65+A is a useful measure that does not assess a homogeneous area but instead different aspects of health and psychosocial functioning. PMID- 19553092 TI - Social and occupational functioning impairment in patients in partial versus complete remission of a major depressive disorder episode. A six-month prospective epidemiological study. AB - PURPOSE: To evaluate social and occupational functioning in patients in partial remission (PR) compared with patients in complete remission (CR) of a major depressive disorder (MDD) episode. SUBJECTS AND METHODS: This is a six-month prospective study. PR was defined as a score more than 7 and less or equal to 15 in the Hamilton Depression Rating Scale, and CR as less or equal to 7. All patients had been on acute antidepressant treatment during the previous three months and no longer met criteria for MDD. Functioning was assessed by the Social and Occupational Functioning Assessment Scale (SOFAS). RESULTS: Mean (S.D.) patient age was 50.5 (14.5) years (N=292) and 77% were female. At baseline, partial remitters showed greater impairment in social and occupational functioning than complete remitters (62.8 [12.6] versus 80.4 [10.5], respectively; P<.0001). After six months, only 47% PR versus 77% CR reached normal functioning, and SOFAS ratings for PR were below normal range (76.2 [12.3] PR versus 84.6 [9.4] CR; P<.0001). PR reported three times more days absent from work due to sickness than CR (63 days versus 20 days; P<.001). CONCLUSION: We conclude that PR of an MDD episode is associated with significant functional impairment that persists even after nine months of antidepressant treatment. Our results underline the importance of treating the patient until achieving full remission. PMID- 19553093 TI - Efficiency of succinylated-olive stone biosorbent on the removal of cadmium ions from aqueous solutions. AB - Chemical functionalization of olive stone wastes with succinate linkers can potentially improve the performance of wastewater treatment technologies via enhanced adsorption and high affinity of the covalently attached succinate groups for heavy metals. In this study, a novel reusable adsorbent material based on agricultural waste has been synthesized by esterifying the lignocellulosic matrix of olive stones with succinic anhydride in toluene under basic conditions. Characterization of the as-prepared material by FTIR and solid-state MAS (13)C NMR spectroscopies and TGA confirmed that the heterogeneous esterification has proceeded very efficiently to yield the succinylated-olive stone (S-OS). Subsequent alkaline treatment of S-OS with saturated NaHCO(3) aqueous solution led to the resulting sodic material (NaS-OS), which was subjected to batch experiments in order to evaluate its cadmium-removing efficiency from aqueous solutions at realistic concentrations of cadmium found in industrial effluents. The results obtained from the sorption characteristics have revealed that NaS-OS material is highly effective in removing cadmium from aqueous solutions, with a maximum uptake capacity of 200 mg g(-1) (1.78 mmol g(-1)). The Langmuir isotherm model was found to fit adequately the equilibrium isotherm data. Cadmium adsorption occurs rapidly and the adsorption mechanism is a chemical sorption via ionic exchange between the adsorbate and adsorbent. Thermodynamic parameters were also evaluated from the effect of temperature studies. Regenerability of NaS-OS material was ascertained by quantitative desorption of cadmium with 1M aqueous NaCl and the reusability of the matrix after five repeated cycles led to nearly no attenuation in its performance (less than 2% in the sorption capacity), indicating that repeated use of NaS-OS is quite feasible. Compared to other low cost adsorbents utilized for the removal of Cd(II) from water/wastewater, NaS-OS shows higher sorption capacity. These results have important implications for the design of low-cost adsorbents based on agricultural wastes. PMID- 19553094 TI - Dietary carbohydrate modification alters serum metabolic profiles in individuals with the metabolic syndrome. AB - BACKGROUND AND AIMS: Whole-grain cereals and diets with a low glycemic index may protect against the development of type 2 diabetes and heart disease, but the mechanisms are poorly understood. We studied the effect of carbohydrate modification on serum metabolic profiles, including lipids and branched chain amino acids, and dependencies between these and specific gene expression pathways in adipose tissue. METHODS AND RESULTS: Twenty subjects with metabolic syndrome were selected from the larger FUNGENUT study population, randomized either to a diet high in oat and wheat bread and potato (OWP) or rye bread and pasta (RP). Serum metabolomics analyses were performed using ultra-performance liquid chromatography coupled to electrospray ionization mass spectrometry (UPLC/MS), gas chromatography (GC) and UPLC. In the OWP group multiple proinflammatory lysophosphatidylcholines increased, while in the RP group docosahexaenoic acid (DHA 22:6n-3) increased and isoleucine decreased. mRNA expression of stress reactions- and adipose tissue differentiation-related genes were up-regulated in adipose tissue in the OWP group. In the RP group, however, pathways related to stress reactions and insulin signaling and energy metabolism were down-regulated. The lipid profiles had the strongest association with the changes in the adipose tissue differentiation pathway when using the elastic net regression model of the lipidomic profiles on selected pathways. CONCLUSION: Our results suggest that the dietary carbohydrate modification alters the serum metabolic profile, especially in lysoPC species, and may, thus, contribute to proinflammatory processes which in turn promote adverse changes in insulin and glucose metabolism. PMID- 19553095 TI - Benefit of Satureja khuzestanica in subchronically rat model of cyclophosphamide induced hemorrhagic cystitis. AB - Cyclophosphamide (CP) as a widely used antineoplastic drug causes hemorrhagic cystitis (HC) mainly via induction of oxidative stress. Regarding established antioxidant potential of Satureja khuzestanica (Lamiaceae) essential oil (SKEO), we aimed to investigate its protective effects in a subchronic rat model of CP induced HC. CP (6mg/kg/day) and SKEO (225mg/kg/day) were administered alone or in combination by gavage for 28 days. Histopathological changes were investigated by light microscopy. Plasma samples were assayed for lipid peroxidation and total antioxidant power as biomarkers of toxic stress. In the CP-treated animals, irregular mucus layer, severe hemorrhage and edema, infiltration of inflammatory cells, and accumulation of mast cells were observed. In the CP+SKEO group, a relatively normal urothelial topography with decreased number of mucosal mast cells and inflammatory cells were observed. Increased lipid peroxidation along with decreased total antioxidant capacity resulting from CP treatment was significantly recovered by SKEO co-treatment. It is concluded that SKEO protects rats from CP-induced HC by reduction of free radical-induced toxic stress. It is strongly recommended to examine SKEO in the clinic to approve its benefit in patients undertaking CP. PMID- 19553096 TI - Quantitative and chemical fingerprint analysis for quality control of rhizoma Coptidischinensis based on UPLC-PAD combined with chemometrics methods. AB - To control the quality of rhizoma Coptidis, a method based on ultra performance liquid chromatography with photodiode array detector (UPLC-PAD) was developed for quantitative analysis of five active alkaloids and chemical fingerprint analysis. In quantitative analysis, the five alkaloids showed good regression (R > 0.9992) within test ranges and the recovery of the method was in the range of 98.4 100.8%. The limit of detections and quantifications for five alkaloids in PAD were less than 0.07 and 0.22 microg/ml, respectively. In order to compare the UPLC fingerprints between rhizoma Coptidis from different origins, the chemometrics procedures, including similarity analysis (SA), hierarchical clustering analysis (HCA), principal component analysis (PCA) were applied to classify the rhizoma Coptidis samples according to their cultivated origins. Consistent results were obtained to show that rhizoma Coptidis samples could be successfully grouped in accordance with the province of origin. Furthermore, five marker constituents were screened out to be the main chemical marker, which could be applied to accurate discrimination and quality control for rhizoma Coptidis by quantitative analysis. This study revealed that UPLC-PAD method was simple, sensitive and reliable for quantitative and chemical fingerprint analysis, moreover, for the quality evaluation and control of rhizoma Coptidis. PMID- 19553097 TI - Utilization of waste tire rubber in manufacture of oriented strandboard. AB - Some physical and mechanical properties of oriented strandboards (OSBs) containing waste tire rubber at various addition levels based on the oven-dry strand weight, using the same method as that used in the manufacture of OSB. Two resin types, phenol-formaldehyde (PF) and polyisocyanate, were used in the experiments. The manufacturing parameters were: a specific gravity of 0.65 and waste tire rubber content (10/90, 20/80 and 30/70 by wt.% of waste tire rubber/wood strand). Average internal bond values of PF-bonded OSB panels with rubber chips were between 17.6% and 48.5% lower than the average of the control samples while polyisocyanate bonded OSBs were 16.5-50.6%. However, water resistance and mechanical properties of OSBs made using polyisocyanate resin were found to comply with general-purpose OSB minimum property requirements of EN 300 Type 1 (1997) values for use in dry conditions at the lowest tire rubber loading level (10%) based on the oven-dry panel weight. The tire rubber improved water resistance of the OSB panel due to its almost hydrophobic property. Based on the findings obtained from this study, we concluded that waste tire rubber could be used for general-purpose OSB manufacturing up to 10% ratio based on the oven-dry panel weight. PMID- 19553098 TI - Development of a method to evaluate caspase-3 activity in a single cell using a nanoneedle and a fluorescent probe. AB - A method to detect an enzymatic reaction in a single living cell using an atomic force microscope equipped with an ultra-thin needle (a nanoneedle) and a fluorescent probe molecule was developed. The nanoneedle enables the low-invasive delivery of molecules attached onto its surface directly into a single cell. We hypothesized that an enzymatic reaction in a cell could be profiled by monitoring a probe immobilized on a nanoneedle introduced into the cell. In this study, a new probe substrate (NHGcas546) for caspase-3 activity based on fluorescent resonance energy transfer (FRET) was constructed and fixed on a nanoneedle. The NHGcas546-modified nanoneedle was inserted into apoptotic cells, in which caspase 3 is activated after apoptosis induction, and a change in the emission spectrum of the immobilized probe could be observed on the surface of the nanoneedle. Thus, we have developed a successful practical method for detecting a biological phenomenon in a single cell. We call the method MOlecular MEter with Nanoneedle Technology (MOMENT). PMID- 19553099 TI - The development and in vitro characterisation of an intracellular nanosensor responsive to reactive oxygen species. AB - Advances in sensor technologies have enhanced our understanding of the roles played by reactive oxygen species (ROS) in a number of physiological and pathological processes. However, high inter-reactivity and short life spans has made real-time monitoring of ROS in cellular systems challenging. Fluorescent dyes capable of intracellular ROS measurements have been reported. However, these dyes are known to be intrinsically cytotoxic and thus can potentially significantly alter cellular metabolism and adversely influence in vitro data. Reported here is the development and in vitro application of a novel ROS responsive nanosensor, based on PEBBLE (Probes Encapsulated By Biologically Localised Embedding) technology. The ROS sensitive fluorescent probe dihydrorhodamine 123 (DHR 123) was employed as the sensing element of the PEBBLE through entrapment within a porous, bio-inert polyacrylamide nanostructure enabling passive monitoring of free radical flux within the intracellular environment. Successful delivery of the nanosensors into NR8383 rat alveolar macrophage cells via phagocytosis was achieved. Stimulation of PEBBLE loaded NR8383 cells with phorbol-12-myristate-13-acetate (PMA) enabled real time monitoring of ROS generation within the cell without affecting cellular viability. These data suggest that PEBBLE nanosensors could offer significant advantages over existing technologies used in monitoring the intracellular environment. PMID- 19553100 TI - Comparison of surface plasmon resonance and capacitive immunosensors for cancer antigen 125 detection in human serum samples. AB - This paper presents a comparison between surface plasmon resonance (SPR) and capacitive immunosensors for a flow injection label-free detection of cancer antigen 125 (CA 125) in human serum. Anti-CA 125 was immobilized on gold surface through a self-assembled monolayer. Parameters affecting the responses of each system were optimized. Under optimal conditions, SPR provided a detection limit of 0.1 U ml(-1) while 0.05 U ml(-1) was obtained for the capacitive system. Linearity for SPR was between 0.1 and 40 U ml(-1) and 0.05-40 U ml(-1) for capacitive system. These immunosensors were applied to analyze CA 125 concentrations in human serum samples and compared with conventional enzyme linked fluorescent assay (ELFA). Both systems showed good agreement with ELFA (P<0.05). Moreover, these immunosensors were very stable and provided good reproducible responses after regeneration, up to 32 times for SPR and 48 times for capacitive system with relative standard deviation lower than 4%. The SPR immunosensor provided advantages in term of fast response and real-time monitoring while capacitive immunosensor offered a sensitive and cost-effective method for CA 125 detection. PMID- 19553101 TI - Biofuels and the conundrum of sustainability. AB - Sustainable energy is the problem of the 21st century. If biofuels want to be part of the solution they must accept a degree of scrutiny unprecedented in the development of a new industry. That is because sustainability deals explicitly with the role of biofuels in ensuring the well-being of our planet, our economy, and our society both today and in the future. Life cycle assessment (LCA) has been the standard framework for assessing sustainability of biofuels. These assessments show that corn ethanol has a marginally lower fossil energy and greenhouse gas footprint compared to petroleum fuel. Sugarcane ethanol and some forms of biodiesel offer substantially lower footprints. New biofuels may offer low footprints. The science of LCA is being stretched to its limits as policy makers consider direct and indirect effects of biofuels on global land and water resources, global ecosystems, air quality, public health, and social justice. PMID- 19553102 TI - Comparative metacognition. AB - Metacognition is knowledge about knowledge, often expressed as confidence judgments about what we know. Most of the literature on metacognition in humans is based on subjects' verbal reports. Investigators of animal cognition have recently described nonverbal methods for investigating metacognition in animals. In one, subjects are given the option to escape from difficult trials. In another, subjects are trained to place bets about the accuracy of their most recent response. To rule out noncognitive interpretations of purported evidence of metacognition in animals, one must ensure that escape responses do not increase the overall density of reinforcement and that they do not occur in the presence of the stimuli on which the subject was trained. The nonverbal techniques used to investigate metacognition in animals make possible two interesting lines of research: investigating the contribution of language and explicit instruction in establishing metacognition, and the investigation of the neural substrates of metacognition. PMID- 19553103 TI - Clinical and histopathological risk factors to predict sentinel lymph node positivity, disease-free and overall survival in clinical stages I-II AJCC skin melanoma: outcome analysis from a single-institution prospectively collected database. AB - BACKGROUND: To investigate if the tumour infiltrating lymphocytes (TILs) are able to predict the sentinel lymph node (SLN) positivity, the disease-free survival (DFS) and overall survival (OS) in clinical stages I-II AJCC primary cutaneous melanoma (PCM). METHODS: The study included consecutive patients with PCM, all diagnosed, treated and followed up prospectively. Logistic regression was used to investigate the association between DFS, OS, SLN positivity and Breslow thickness, Clark level, TIL, ulceration, lesion site, gender, regression and age. RESULTS: From November 1998 to October 2008, 1251 consecutive patients with PCM were evaluated. Median age was 51 (range 15-96) with 32.2% (N=393) of them older than 60; 44.8% of them were males. Of the whole series, a total of 404 patients with primary vertical growth phase (VGP) melanoma and no clinical evidence of metastatic disease underwent SLN biopsy. Of these, 74 (18.8%) had a positive SLN. In a multivariate analysis, primary melanoma on the extremities versus that on the axial locations (truncal and head/neck) (OR 0.49, 95% CI 0.25-0.98, p 0.04) and TILs (TILs versus no TILs) (OR 0.47, 95%CI 0.25-0.90, p 0.02) were predictive for lower probability of SLN involvement, while thickness (>4mm versus 0-1mm) (OR 24, 19, 95% CI 4.91-119.13, p<.001) was predictive for higher risk of SLN positivity. A multivariate stepwise analysis confirmed these results. The histological status of the SLN was the most significant predictor of DFS and OS. Patients with a negative SLN had a 5-year DFS of 75.9%, compared with 35.2% in patients with a positive SLN (p<.0001) and a 5-year OS of 88.7% versus 42.9%, respectively (p<.0001). CONCLUSIONS: Our study demonstrates that the absence of TILs predicts SLN metastasis, in multivariate analysis the SLN positivity predicts DFS and OS. PMID- 19553104 TI - Cell cycle and apoptotic effects of SAHA are regulated by the cellular microenvironment in HCT116 multicellular tumour spheroids. AB - Using multicellular tumour spheroids (MCTS) of HCT116 colon carcinoma cells, we analysed the effects of SAHA (suberoylanilide hydroxamic acid), a histone deacetylase inhibitor (HDACi). We found that, although SAHA-induced histone acetylation and ROS level upregulation occur throughout the spheroid, inhibition of cell cycle progression and induction of apoptosis are dependent on cell microenvironment. SAHA-induced growth inhibition of HCT116 MCTS results from the inhibition of cell cycle progression and induction of apoptosis. At a low concentration SAHA decreases Ki-67 and cyclin A positive cells and increases p21 positive cells in the outer layer while it induces a ROS-dependent apoptosis in the central zone of the spheroid. Coimmunostaining of p21 and apoptotic cells confirms that SAHA effects are different depending on the position of the cells within the spheroid. At a higher dose, SAHA induces mitotic defects and survivin downregulation in the outer layer of cells resulting in an additional cytotoxic effect in this part of the spheroid. Together these findings show that SAHA induced cytostatic and cytotoxic effects occur in different cell populations, indicating that the cellular microenvironment is an important determinant in the regulation of the effects of SAHA treatment. Consequently, the MCTS model appears to be a valuable advanced tool for evaluating the effects of SAHA treatment in combination with other anticancer agents. PMID- 19553105 TI - Evaluation and modeling of biochemical methane potential (BMP) of landfilled solid waste: a pilot scale study. AB - The main goal of this study was to present a comparison of landfill performance with respect to solids decomposition. Biochemical methane potential (BMP) test was used to determine the initial and the remaining CH(4) potentials of solid wastes during 27 months of landfilling operation in two pilot scale landfill reactors. The initial methane potential of solid wastes filled to the reactors was around 0.347 L/CH(4)/g dry waste, which decreased with operational time of landfill reactors to values of 0.117 and 0.154 L/CH(4)/g dry waste for leachate recirculated (R1) and non-recirculated (R2) reactors, respectively. Results indicated that the average rate constant increased by 32% with leachate recirculation. Also, the performance of the system was modeled using the BMP data for the samples taken from reactors at varying operational times by MATLAB program. The first-order rate constants for R1 and R2 reactors were 0.01571 and 0.01195 1/d, respectively. The correlation between the model and the experimental parameters was more than 95%, showing the good fit of the model. PMID- 19553106 TI - Life cycle assessment of biofuels: energy and greenhouse gas balances. AB - The promotion of biofuels as energy for transportation in the industrialized countries is mainly driven by the perspective of oil depletion, the concerns about energy security and global warming. However due to sustainability constraints, biofuels will replace only 10 to 15% of fossil liquid fuels in the transport sector. Several governments have defined a minimum target of GHG emissions reduction for those biofuels that will be eligible to public incentives, for example a 35% emissions reduction in case of biofuels in Members States of the European Union. This article points out the significant biases in estimating GHG balances of biofuels stemming from modelling choices about system definition and boundaries, functional unit, reference systems and allocation methods. The extent to which these choices influence the results is investigated. After performing a comparison and constructive criticism of various modelling choices, the LCA of wheat-to-bioethanol is used as an illustrative case where bioethanol is blended with gasoline at various percentages (E5, E10 and E85). The performance of these substitution options is evaluated as well. The results show a large difference in the reduction of the GHG emissions with a high sensitivity to the following factors: the method used to allocate the impacts between the co products, the type of reference systems, the choice of the functional unit and the type of blend. The authors come out with some recommendations for basing the estimation of energy and GHG balances of biofuels on principles such as transparency, consistency and accuracy. PMID- 19553107 TI - Identification of aminoethyl pyrrolo dihydroisoquinolinones as novel poly(ADP ribose) polymerase-1 inhibitors. AB - PARP inhibitors have been demonstrated to retard intracellular DNA repair and therefore sensitize tumor cells to cytotoxic agents or ionizing radiation. We report the identification of a novel class of PARP1 inhibitors, containing a pyrrolo moiety fused to a dihydroisoquinolinone, derived from virtual screening of the proprietary collection. SAR exploration around the nitrogen of the aminoethyl appendage chain of 1 led to compounds that displayed low nanomolar activity in a PARP1 enzymatic assay. PMID- 19553108 TI - Discovery and structural analysis of Eph receptor tyrosine kinase inhibitors. AB - The Eph family of receptor tyrosine kinases has drawn growing attention due to their role in regulating diverse biological phenomena. However, pharmacological interrogation of Eph kinase function has been hampered by a lack of potent and selective Eph kinase inhibitors. Here we report the discovery of compounds 6 and 9 using a rationally designed kinase-directed library which potently inhibit Eph receptor tyrosine kinases, particularly EphB2 with cellular EC(50)s of 40nM. Crystallographic data of EphA3 and EphA7 in complex with the inhibitors show that they bind to the 'DFG-out' inactive kinase conformation and provide valuable information for structure-based design of second generation inhibitors. PMID- 19553109 TI - Development of enzymatically cleavable prodrugs derived from dendritic polyglycerol. AB - In this Letter we report the synthesis and in vitro studies of cleavable polymer drug conjugates derived from dendritic polyglycerol and maleimide-bearing prodrugs of doxorubicin and methotrexate that are cleaved by cathepsin B. Cleavage properties and cytotoxicity of the new conjugates are presented. PMID- 19553110 TI - Development of a selective and potent radioactive ligand for histamine H(3) receptors: A compound potentially useful for receptor occupancy studies. AB - Radioligands are powerful tools for examining the pharmacological profiles of chemical leads and thus facilitate drug discovery. In this study, we identified and characterized 3-([1,1,1-(3)H]methyl)-2-(4-{[3-(1-pyrrolidinyl)propyl]oxy} phenyl)-4(3H)-quinazolinone ([(3)H]1) as a potent and selective radioligand for histamine H(3) receptors. Radioligand [(3)H]1 exhibited appreciable specific signal in brain slices prepared from wild-type mice but not from histamine H(3) receptor-deficient mice, demonstrating the specificity and utility of [(3)H]1 as a selective histamine H(3) receptor radioligand for ex-vivo receptor occupancy assays. PMID- 19553111 TI - Amidine derived inhibitors of acid-sensing ion channel-3 (ASIC3). AB - A series of indole amidines modified at the 2-position of the indole ring were evaluated as inhibitors of Acid-Sensing Ion Channel-3 (ASIC3), a novel target for the treatment of chronic pain. PMID- 19553112 TI - Discovery and optimization of a novel Neuromedin B receptor antagonist. AB - The discovery and parallel synthesis of potent, small molecule antagonists of Neuromedin B receptor based on the ary-hexahydro-dibenzodiazepin-1-one core is described. PMID- 19553113 TI - Dopamine D(3) receptor antagonists: The quest for a potentially selective PET ligand. Part two: Lead optimization. AB - The lead optimization process to identify new selective dopamine D(3) receptor antagonists is reported. DMPK parameters and binding data suggest that selective D(3) receptor antagonists as potential PET ligands might have been identified. PMID- 19553114 TI - Synthesis and SAR of novel tricyclic quinoxalinone inhibitors of poly(ADP ribose)polymerase-1 (PARP-1). AB - Based on screening hit 1, a series of tricyclic quinoxalinones have been designed and evaluated for inhibition of PARP-1. Substitutions at the 7- and 8-positions of the quinoxalinone ring led to a number of compounds with good enzymatic and cellular potency. The tricyclic quinoxalinone class is sensitive to modifications of both the amine substituent and the tricyclic core. The synthesis and structure activity relationship studies are presented. PMID- 19553115 TI - Brain-penetrating 2-aminobenzimidazole H(1)-antihistamines for the treatment of insomnia. AB - The benzimidazole core of the selective non-brain-penetrating H(1)-antihistamine mizolastine was used to identify a series of brain-penetrating H(1) antihistamines for the potential treatment of insomnia. Using cassette PK studies, brain-penetrating H(1)-antihistamines were identified and in vivo efficacy was demonstrated in a rat EEG/EMG model. Further optimization focused on strategies to attenuate an identified hERG liability, leading to the discovery of 4i with a promising in vitro profile. PMID- 19553116 TI - Severe nemaline myopathy associated with consecutive mutations E74D and H75Y on a single ACTA1 allele. AB - Nemaline myopathy is among the most common congenital myopathies. We describe for the first time a novel double de novo mutation in two adjacent codons resulting in two amino acid changes E74D and H75Y in the ACTA1 gene. The hypotonic male infant was the first son of healthy unrelated parents with no family history of neuromuscular disorders. Pregnancy was complicated: decreased fetal movements were noted on the 25th week of gestation, premature labour pains were present from the 29th week onwards and because of breech presentation a Caesarian section was carried out in the 39th week. The patient presented with multiple congenital fractures and joint contractures. He was dependent on ventilatory support until his death at 2 months. Muscle biopsy revealed severely atrophic and rounded muscle fibers with considerable variation in diameter and pronounced disorganization of the myofibers. Electron microscopy indicated a distinct disturbance of the myofibrillar architecture and nemaline rods. In view of previously described cases carrying different single missense mutations of the amino acid residues E74 or H75, we suggest that the particular genotype E74D/H75Y is compatible with the severity of the patient's phenotype. The possibility of germ cell mosaicism should be taken into account in genetic counseling. PMID- 19553118 TI - A TPM3 mutation causing cap myopathy. AB - Cap disease is a rare congenital myopathy associated with skeletal malformations and respiratory involvement. Abnormally arranged myofibrils taking the appearance of a "cap" are the morphological hallmark of this entity. We report a case of cap disease concerning a 42-year-old man, without any family history and presenting a p.Arg168His mutation on the TPM3 gene. His first biopsy at 7years had only shown selective type I hypotrophy. Mutations of TPM3 gene have been found in nemaline myopathy, congenital fiber type disproportion, but never before in cap disease. PMID- 19553119 TI - On the spectrum of leprosy neuropathies: multifocal inflammatory neuropathy heralding leprosy relapse. AB - We report a 52 year-old woman with a past history of lepromatous leprosy (14 years prior to our first evaluation) who presented with progressive weakness and severe arm/leg pain. CSF analysis revealed elevated protein level with normal cell count. Skin and sural nerve biopsy showed no bacilli. Immunomodulatory treatment led to major improvement on clinical, CSF and electrodiagnostic grounds, but after one year of treatment, skin test revealed leprosy relapse. To our knowledge, this is the first report of a multifocal inflammatory neuropathy heralding leprosy relapse. Extended neurological work-up may be important in unexplained neuropathy progression after leprosy treatment. PMID- 19553122 TI - Contribution of seat and foot reaction forces to anticipatory postural adjustments (APAs) in sitting isometric ramp pushes. AB - The aim of this paper was to examine the role of the upper and lower body on the dynamic phenomena, which precede the voluntary movement (anticipatory postural adjustments: APAs), and the way in which they contribute to postural control. In this view, sitting subjects were asked to perform horizontal two-handed ramp pushes as quickly as possible. A dynamometric bar was used to provide the push force (F(x)). Local reaction forces along the antero-posterior and vertical axes, at the seat and foot-rests (R(Sx), R(Sz), and R(fx), R(fz), respectively), as well as global ones (R(x) and R(z)), were measured. Two postural conditions were considered: full (100 BP) and one-third ischio-femoral contact (30 BP). Anticipatory postural adjustments durations (dAPAs) were measured between the onset of global or local (that is, at the seat and foot level) reaction forces, and the onset of push force increase. Firstly, the dAPAs were longer at the foot than at the seat level, that is, the APA sequence starts at the foot level: it is suggested that a "posturo-focal" sequence is followed, whose progression order is precisely dependent on the postural conditions. Moreover, the APA peak amplitudes (pAPA), measured at the seat contact were significantly greater than the corresponding ones measured at the foot contact: the upper body dynamics are larger than the lower body dynamics. Secondly, a greater peak push force (pF(x)) entailed significant dAPA increases, in preference to pAPA increases. As APAs are dynamic phenomena, they can perturb balance, suggesting that, in order to avoid unnecessary perturbation, APAs are increased in terms of duration rather than amplitude. Lastly, the impulses corresponding to the push force increase ("BPI(x)") and to the APA periods ("ACPI(x)") were calculated. As ACPI(x) was very low as compared to BPI(x), it was suggested that the APA action was limited to the period of the voluntary movement onset. PMID- 19553120 TI - Reliability of the North Star Ambulatory Assessment in a multicentric setting. AB - The aim of this study was to investigate the suitability of the North Star Ambulatory Assessment as a possible outcome measure in multicentric clinical trials. More specifically we wished to investigate the level of training needed for achieving a good interobserver reliability in a multicentric setting. The scale was specifically designed for ambulant children with Duchenne Muscular Dystrophy and includes 17 items that are relevant for this cohort. Thirteen Italian centers participated in the study. In the first phase of the study we provided two training videos and an example of the scale performed on a child. After the first session of training, all the 13 examiners were asked to send a video with an assessment performed in their centre and to score all the videos collected. There were no difficulties in performing the items and in obtaining adequate videos with a hand held camera but the results showed a poor interobserver reliability (<.5). After a second training session with review and discussion of the videos previously scored, the same examiners were asked to score three new videos. The results of this session had an excellent interobserver reliability (.995). The level of agreement was maintained even when the same videos were rescored after a month, showing a significant intra-observer reliability (.95). Our results suggest that the NSAA is a test that can be easily performed, completed in 10 min and can be used in a multicentric setting, providing that adequate training is administered. PMID- 19553121 TI - Nemaline (actin) myopathy with myofibrillar dysgenesis and abnormal ossification. AB - We report a 2-year-old boy who presented with marked hypotonia and was dependent on artificial ventilation since birth. He was diagnosed with nemaline (actin) myopathy, based on the cytoplasmic accumulation of thin filament aggregates and marked myofibrillar dysgenesis. Intranuclear rods and dispersed tiny nemaline bodies were also observed. The patient was shown to be heterozygous for a de novo mutation, c.430C>T (p.Leu144Phe), in the alpha-actin (ACTA1) gene. He also showed orbital osteosclerosis, longitudinal striations of the iliac bones, hepatomegaly, undescended testis, a unilateral vesico-ureteric stenosis, severe failure to thrive, and dilatation of the lateral cerebral ventricles. Besides the severe muscle involvement, these clinical findings further broaden the clinical spectrum of actinopathy phenotypes. PMID- 19553123 TI - Increased shoe sole hardness results in compensatory changes in the utilized coefficient of friction during walking. AB - Based on mechanical testing, harder soled shoes have been shown to provide less available friction than soft soled shoes. Whether or not humans adjust their utilized coefficient of friction (COFu) and gait kinematics to accommodate the decrease in available friction while wearing hard soled shoes is not known. Fifty six young adults participated in this study. Ground reaction forces, full body kinematics, stride characteristics and subjective perception of footwear slipperiness were recorded under both hard and soft soled shoe conditions. Paired t-tests were used to identify the differences between two shoes conditions. Results indicated that the peak COFu was significantly less when wearing the hard soled shoes compared to when wearing the soft soled shoes (0.23 vs. 0.26, P<0.001). The decrease in peak COFu was the result of a decrease in the resultant shear forces at the time of peak COFu as no difference in the vertical ground reaction forces was observed. When wearing hard soled shoes, subjects demonstrated decreased total body center of mass (COM) acceleration prior to and immediately following initial contact, decreased walking velocity, shortened stride length, and reduced ankle dorsiflexion angle at initial contact. Taken together, we believe that these gait modifications represent behavioral adaptations to wearing shoes that are perceived to be more slippery. PMID- 19553124 TI - An unusual stab wound causing a traumatic pseudomeningocele at the craniocervical junction. AB - A stab wound to the craniocervical junction complicated by a pseudomeningocele has been reported rarely. A 65-year-old man was stabbed with a knife, and the blade penetrated the space between the occipital bone and the atlas. On admission, he presented with quadriparesis; the left arm was predominantly affected. The sensory level was at C2 on the left side. T2-weighted MRI showed a focal area of hyperintensity at the cervicomedullary junction, localized to the left of the spinal cord. The wound was carefully cleaned and closed. Follow-up MRI revealed a large pseudomeningocele along the trajectory, which resorbed completely without intervention. The quadriparesis improved progressively, except for the left arm, which improved only slightly. The asymptomatic insignificant meningocele completely improved spontaneously without surgery. PMID- 19553125 TI - Dopamine dysregulation syndrome, impulse control disorders and punding after deep brain stimulation surgery for Parkinson's disease. AB - Data regarding the effect of deep brain stimulation (DBS) surgery on the dopamine dysregulation syndrome (DDS), impulse control disorders (ICDs) and punding in Parkinson's disease (PD) are limited. We present a case series of 21 operated PD patients who had exhibited DDS, ICDs or punding at some stage during the disease. DDS remained unimproved or worsened post-operatively in 12/17 patients with pre operative DDS (71%) (nine bilateral subthalamic nucleus [STN], one right-sided STN, two bilateral globus pallidus internus [GPi] DBS). DDS improved or resolved after bilateral STN DBS in 5/17 patients with pre-operative DDS. DDS apparently developed for the first time after bilateral STN DBS in two patients, although only after a latency of eight years in one case. One patient without reported pre operative DDS or ICDs developed pathological gambling post-STN DBS. One patient had pathological gambling which resolved pre-operatively, and did not recur post DBS. Thus, DDS, ICDs and punding may persist, worsen or develop for the first time after DBS surgery, although a minority of patients improved dramatically. Predictive factors may include physician vigilance, motor outcome and patient compliance. PMID- 19553126 TI - Single-level bilateral facet joint hypertrophy causing thoracic spinal canal stenosis. AB - Thoracic canal stenosis caused by hypertrophy of the posterior spinal elements is rare. We report an unusual case of bilateral zygapophyseal joint hypertrophy occurring solely at thoracic levels T10-11, producing bilateral leg weakness and numbness. The diagnosis was established using CT scans and MRI. A wide decompressive laminectomy and posterolateral bone fusion was performed. Postoperatively there was a marked improvement in symptoms and signs. We present a literature review on bilateral single-level facet joint hypertrophy producing canal stenosis and report our experience. PMID- 19553127 TI - Giant intradural extramedullary arachnoid cyst of the thoracic spine. AB - Spinal intradural arachnoid cysts (ACs) are found frequently in the thoracic region, and often extend over four or five vertebral levels. We present a 28-year old patient who had a giant thoracic congenital intradural extramedullary AC (T1 T12) with a 10-month history of pain, paresthesia, paraparesis and gait ataxia. A T3 to T6 laminectomy was performed. After durotomy, the posterior wall of the AC was visualized compressing the spinal cord. We resected the cyst wall as widely as possible and connected the cyst to the subarachnoid space using a catheter. There were no postoperative complications. At 1-year follow-up, the patient presented with no motor deficits or pain, and had experienced progressive resolution of the gait ataxia. The treatment of giant intradural extramedullary ACs, especially for those that cannot be totally excised, should include generous fenestration and the insertion of a cyst-subarachnoid shunt. PMID- 19553128 TI - Early failure of a unicompartmental knee arthroplasty design with an all polyethylene tibial component. AB - Refined prosthetic designs and surgical techniques for unicompartmental knee arthroplasty have recently been associated with improved outcomes. The purpose of the present study was to evaluate the clinical and radiographic outcomes of the EIUS unicompartmental design, which has an all-polyethylene tibial component, and to compare these outcomes with published reports of other unicompartmental prostheses. Between February 2002 and March 2005, 113 patients (144 knees) underwent a medial unicompartmental knee arthroplasty, all performed by a single surgeon who used the EIUS prosthesis. At a mean follow-up of 36 months (range, 24 54 months), the mean Knee Society objective and functional scores improved from 55 points (range, 31-77 points) and 49 points (range, 35-60 points) to 92 points (range, 45-100 points) and 89 points (range, 10-100 points), respectively. The implant survival rate was 89%, with 16 knees either revised or scheduled for revision. The reasons for revision included aseptic loosening of the tibial component (eight knees), progressive symptomatic patellofemoral disease (four knees), and tibial component subsidence (four knees). Multiple regression analysis revealed that age, gender, and body mass index were not significantly correlated with success or failure of this design, although nine of the 16 patients who required revision were obese. This prosthesis was associated with higher revision rates than components which utilize metal-backed implants. Further modifications in the design, indications, or technique may be necessary to improve outcomes of this unicompartmental knee arthroplasty system. PMID- 19553130 TI - Comparison of two full automatic synthesis methods of 9-(4-[(18)F]fluoro-3 hydroxymethylbutyl)guanine using different chemistry modules. AB - We have developed synthesis methods for 9-(4-[(18)F]fluoro-3 hydroxymethylbutyl)guanine ([(18)F]FHBG) using two commercial automatic chemistry modules, Tracerlab MX and Explora RN, and compared radiochemical yields. Synthesis conditions and sequence programs were modified for two modules because both these modules have different mechanical structures, including heater type, vacuum system, reactor, and tubing size. Synthesis using the Tracerlab MX module showed a 21.0+/-3.8% yield of radiochemical, which was 98+/-0.9% pure; the total preparation time was 63.0+/-5.0min including an HPLC purification step. In contrast, synthesis using the Explora RN module showed a 32.0+/-1.2% yield of radiochemical, which was 99.0+/-0.6% pure; the total preparation time was 38+/ 2min, using different HPLC purification conditions and without the HPLC solvent evaporation step. PMID- 19553131 TI - Development and calibration of a gamma-ray density-meter for sediment-like materials. AB - The core-logger PHAROS was designed at Kernfysisch Versneller Instituut, Groningen, the Netherlands to measure activity concentration of sediment like materials. As the core logger was calibrated for a single material density the variations in the density profile in the scanned core can result in erroneous estimations of activity concentrations. To overcome this, a density-meter was developed and implemented on PHAROS. The density measurement should be non invasive and should cover a wide range of sediment like materials. The most suitable approach is to deploy gamma-ray transmission technique, where the intensity of non-attenuated photons after traversing the core is directly related to its density. Due to the overall complexity of radiation transport through sediment media, the design of the density-meter was assisted by Monte Carlo simulations. They were deployed to model sophisticated and time-consuming experiments in the process of designing the (137)Cs-based source, to generate the pulse-height response functions of the bismuth-germanate (BGO) scintillator and to estimate the systematic uncertainty induced by the core displacement in the process of the measurement. Moreover, the Monte Carlo simulations have demonstrated as a reliable complementary tool for the generation of PHAROS detector calibration curves. The designed density-meter of PHAROS core-logger indicates to be adequate to estimate densities ranging from approximately 1.6 to approximately 2.7gcm(-3) with the systematic uncertainty within 3%. PMID- 19553129 TI - Inhibitors of VIM-2 by screening pharmacologically active and click-chemistry compound libraries. AB - VIM-2 is an Ambler class B metallo-beta-lactamase (MBL) capable of hydrolyzing a broad-spectrum of beta-lactam antibiotics. Although the discovery and development of MBL inhibitors continue to be an area of active research, an array of potent, small molecule inhibitors is yet to be fully characterized for VIM-2. In the presented research, a compound library screening approach was used to identify and characterize VIM-2 inhibitors from a library of pharmacologically active compounds as well as a focused 'click' chemistry library. The four most potent VIM-2 inhibitors resulting from a VIM-2 screen were characterized by kinetic studies in order to determine K(i) and mechanism of enzyme inhibition. As a result, two previously described pharmacologic agents, mitoxantrone (1,4 dihydroxy-5,8-bis([2-([2-hydroxyethyl]amino)ethyl]amino)-9,10-anthracenedione) and 4-chloromercuribenzoic acid (pCMB) were found to be active, the former as a non-competitive inhibitor (K(i)=K(i)(')=1.5+/-0.2microM) and the latter as a slowly reversible or irreversible inhibitor. Additionally, two novel sulfonyl triazole analogs from the click library were identified as potent, competitive VIM-2 inhibitors: N-((4-((but-3-ynyloxy)methyl)-1H-1,2,3-triazol-5-yl)methyl)-4 iodobenzenesulfonamide (1, K(i)=0.41+/-0.03microM) and 4-iodo-N-((4 (methoxymethyl)-1H-1,2,3-triazol-5-yl)methyl)benzenesulfonamide (2, K(i)=1.4+/ 0.10microM). Mitoxantrone and pCMB were also found to potentiate imipenem efficacy in MIC and synergy assays employing Escherichia coli. Taken together, all four compounds represent useful chemical probes to further investigate mechanisms of VIM-2 inhibition in biochemical and microbiology-based assays. PMID- 19553132 TI - Free flaps: outcomes and complications in head and neck reconstructions. AB - Between 2000 and 2007, 376 free-flap transfers were performed in 352 patients at the Department of Maxillofacial Surgery, University of Parma, Italy. They included 303 reconstructions after cancer surgery (80.6%), 28 secondary reconstructions (7.4%), 32 transfers for facial paralysis (8.5%) and 13 reconstructions for other pathology (3.5%) such as osteonecrosis and ameloblastoma. We determined the rates of major and minor flap, donor-site and systemic complications and analysed their relationships with factors such as aetiology, patient age, smoking history and the presence of comorbid conditions. For the free flaps examined in this study, the overall complication rate was 47%. Major complications occurred in 20.7% of the cases and minor complications in 26.3%. The major flap, donor-site and systemic complication rates were 11.9, 3.2 and 5.6%, respectively. Total flap loss occurred in 15 cases (4%). The minor flap, donor-site and systemic complication rates were 19.1, 4 and 3.2% respectively. Aetiology, patient age, smoking history and the presence of comorbid conditions were related to higher rates of major and minor complications, although these were not statistically significant. PMID- 19553133 TI - Computational principles of determining and improving mass precision and accuracy for proteome measurements in an Orbitrap. AB - Precision proteomics requires high-resolution and high mass accuracy peptide measurements. The Orbitrap instrument achieves excellent resolution on a chromatographic time scale and its design is favorable for very high mass accuracy. Here we describe how mass precision for each peptide increases successively by considering all associated measurements, starting from the MS peak and proceeding to its chromatographic elution profile, isotope envelope, and stable isotope pair in SILAC measurements. We extract peptide charge pairs to perform nonlinear recalibration of the Orbitrap mass scale through spline interpolation. The deviation of mass values determined from charge pairs is used to convert mass precision to mass accuracy for subsequent database search. The corrected mass precision is consistent with the mass accuracy independently determined by database identification. Individual mass deviations range from below 100 ppb for peptides with many associated mass measurements and good signal intensities to low ppm for peptides with few mass measurements and signals close to the noise level. This extremely high and individualized mass accuracy is equivalent to a substantial increase in database identification score. PMID- 19553134 TI - Anterior chest discomfort and right neck pain in a young woman 2 days after an appendectomy. PMID- 19553135 TI - A rounded image inside the left ventricle: the mechanism of the artifact formation. PMID- 19553136 TI - Intra-aortic balloon counterpulsation in an off-pump procedure shows improved cerebral perfusion by near-infrared spectroscopy. PMID- 19553139 TI - Echocardiography in acute native aortic valve endocarditis. PMID- 19553140 TI - Effects of fenoldopam mesylate on central hemodynamics and renal flow in patients undergoing cardiac surgery: color Doppler echocardiographic evaluation. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of 0.1 microg/kg/min of fenoldopam mesylate on renal flow and central hemodynamics measured by echocardiography in hemodynamically stable patients with preserved renal function undergoing cardiac surgery. DESIGN: Experimental observational study. SETTING: Single-institutional community hospital study. PARTICIPANTS: Thirty patients undergoing cardiac surgery. INTERVENTION: Fenoldopam mesylate infusion (0.1 microg/kg/min) in patients undergoing cardiopulmonary bypass. MEASUREMENTS AND MAIN RESULTS: Doppler measurements of renal blood flow and echocardiographic hemodynamic determinations after Doppler echocardiography measured flux velocities of the main, segmental, and interlobar and interlobular right renal arteries. The authors calculated the resistive index of all the renal segments studied. Moreover, the authors measured the flux of the main renal artery and its diameter as well as the main hemodynamic variables. All the measurements were performed in the intensive care unit setting at baseline and 20 minutes after the infusion of 0.1 microg/kg/min of fenoldopam mesylate. Fenoldopam mesylate infusion significantly increased blood flow in all renal compartments, thus improving the resistive index. The study showed that fenoldopam mesylate infusion does not induce any significant change of the heart rate or arterial pressure, cardiac output, preload, or wall stress. CONCLUSIONS: In hemodynamically stable cardiac surgery patients with preserved renal function, an infusion of 0.1 microg/kg/min of fenoldopam mesylate has no influence on systemic hemodynamics while increasing renal blood flow. PMID- 19553141 TI - The mental time line: an analogue of the mental number line in the mapping of life events. AB - A crucial aspect of the human mind is the ability to project the self along the time line to past and future. It has been argued that such self-projection is essential to re-experience past experiences and predict future events. In-depth analysis of a novel paradigm investigating mental time shows that the speed of this "self-projection" in time depends logarithmically on the temporal-distance between an imagined "location" on the time line that participants were asked to imagine and the location of another imagined event from the time line. This logarithmic pattern suggests that events in human cognition are spatially mapped along an imagery mental time line. We argue that the present time-line data are comparable to the spatial mapping of numbers along the mental number line and that such spatial maps are a fundamental basis for cognition. PMID- 19553142 TI - Modulating effect of atorvastatin on paraoxonase 1 activity in type 2 diabetic Egyptian patients with or without nephropathy. AB - The aim of this study was to investigate the modulating effect of atorvastatin on serum paraoxonase 1 enzyme (PON1) activity in type 2 diabetic Egyptian patients with or without nephropathy. The present study was carried out on the following groups: control group, which consisted of 30 healthy persons; Group I, which consisted of 20 type 2 diabetic patients without nephropathy; and Group II, which consisted of 20 type 2 diabetic patients with nephropathy. All the patients selected were under an antidiabetic regimen of insulin, and patients receiving antihypertensive agents were excluded from the follow-up study to avoid drug interaction fallacies. Twenty-two patients (15 without nephropathy and seven with nephropathy) received atorvastatin in individually adjusted oral dosage (range 10 20 mg) once per day for 12 weeks. All cases were subjected to thorough clinical examination and history taking and measurement of serum levels of PON1 activity, malondialdehyde (MDA), glutathione reductase activity, fasting glucose, total cholesterol, triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL), urea, and creatinine. Urine samples were collected for determination of proteinuria. The obtained results showed that PON1 activity and HDL significantly decreased and fasting glucose significantly increased in Group I and Group II when compared to the control group, with significant difference in their levels between Group II and Group I. MDA, total cholesterol, and LDL levels significantly increased and glutathione reductase activity significantly decreased in Group I and Group II when compared to the control group. Urea, creatinine, and proteinuria levels showed significant increase in Group II when compared to the control group and Group I, with nonsignificant difference between control group and Group I. Atorvastatin therapy caused a significant increase in PON1 activity, and serum levels of MDA and glutathione reductase activity were significantly decreased and increased, respectively. Also, total cholesterol, triglyceride and LDL-cholesterol levels were significantly reduced with a significant increase in HDL-cholesterol levels. There was a significant modest reduction in serum urea and creatinine levels as well as in proteinuria level. Fasting glucose level was significantly reduced under the antidiabetic regimen of insulin through the follow-up period. PON1 activity showed a significant negative correlation with glucose and LDL, and a significant positive correlation with HDL in all the studied groups. It could be concluded that atorvastatin with its pleiotropic effects could provide optimal therapeutic intervention to control not only dyslipidemia, but also oxidative stress status with consequent improvement in the course of type 2 diabetes and diabetic nephropathy. More specifically, restoration of PON1 activity by atorvastatin opens a window to investigate other drugs that could provide a new adjuvant therapeutic line for better control of diabetes and diabetic nephropathy. Further research is also recommended to study the distribution of PON1 genetic polymorphism among the Egyptian population to explain the variability in its activity and its relationship with other factors that associate diabetes and its complications. PMID- 19553143 TI - Hemodynamics and rupture of terminal cerebral aneurysms. AB - RATIONALE AND OBJECTIVES: The objective of this study was to investigate the relationship between hemodynamics patterns and aneurysmal rupture in cerebral aneurysms of the same morphology regardless their location. Particularly, terminal aneurysms in both the anterior and posterior circulation were studied. MATERIALS AND METHODS: A total of 42 patient-specific vascular models were constructed from three-dimensional rotational angiography images. All patients had terminal aneurysms at different arteries: a) middle cerebral; b) anterior communicating; c) internal carotid (terminus); d) internal carotid-posterior communicating; e) basilar; or f) anterior cerebral. Hemodynamics information (intra-aneurysmal velocity and wall shear stress distributions) was derived from image-based computational fluid dynamics models with realistic patient-specific anatomies. RESULTS: The group of aneurysms with an inflow jet that splits in two secondary jets, one of which enters the aneurysm before reaching one of the daughter vessels (type B), had the highest peak wall shear stress (WSS) and the highest rupture rate. The peak WSS averaged over each flow type showed a higher value in the ruptured group. The average peak WSS in the ruptured group (all types) was 188 dyn/cm(2) (compared to 118 dyn/cm(2) for the unruptured). CONCLUSIONS: This finding is in agreement with a previous work in which only anterior communicating artery aneurysms were investigated. The significance of these findings is that, if they are statistically confirmed with larger number of cases, flow types could be directly observed during angiographic examinations and linked to WSS categories that may help evaluate which aneurysms are more likely to rupture. PMID- 19553144 TI - Gaucher disease in Tunisia: High frequency of the most common mutations. PMID- 19553145 TI - Not all DMT1 mutations lead to iron overload. AB - DMT1 is a membrane-bound divalent metal transporter, which co-transports protons and (Fe(2+)) from an acidic microenvironment (endosome, duodenal lumen) to the cell cytosol. Results from animal models and from patients have shown that DMT1 is required for intestinal iron absorption and iron acquisition by erythrocytes. Only three human patients with DMT1 mutations have been described so far. They presented with hypochromic microcytic anemia and heavy liver iron overload, even at a very young age. Here, we report the fourth human case, a 7-year old boy with a new homozygous DMT1 mutation, microcytic anemia but no liver iron overload. The mutation introduces a Glycine to Arginine (p.G75R) amino acid substitution. Glycine75 is a highly conserved amino acid present in the first transmembrane domain of the protein and we hypothesize that this mutation fully impairs ferrous iron uptake from the diet and prevents the onset of liver iron overload. PMID- 19553146 TI - Animal poisoning in Europe. Part 2: Companion animals. AB - This is the second in a series of three review articles on animal poisoning in Europe and focuses on cases in pet animals and horses in five European countries (Belgium, France, Greece, Italy and Spain) reported over the last decade. In the participating countries, dogs were the most commonly poisoned species, particularly younger animals. The majority of cases in companion animals resulted from exposure to insecticides, although rodenticides (especially anticoagulants and strychnine) posed a significant risk. In all five countries, horses and cats appeared to be more susceptible to plant toxins. Intoxications with herbicides, metals, household products and drugs for veterinary and human use were reported sporadically. The review demonstrates the importance of increased awareness so as to minimise poisoning episodes and emphasises the need to establish a European system for the recording of poisoning data. PMID- 19553147 TI - Bone marrow biopsy from the flipper of a dolphin. AB - To find macroscopically palpable bone marrow cavities in dolphins is difficult because of their extremely retrogressive limbs and pelvis and because they do not contain abundant modular cavities (as in terrestrial mammals) that can serve as sites for bone marrow biopsies. Three-dimensional computed tomography analysis of dolphin skeletons suggests that bone marrow could be harvested from the humerus and radius. In this report, post-mortem paracentesis of the humerus from a captive rough-toothed dolphin using a biopsy needle provided a marrow preparation containing myelocytes, erythroblasts and megakaryocytes. This type of bone marrow collection from the flipper might be useful for clinical diagnostic work in cetaceans. PMID- 19553150 TI - Intralesional amphotericin B in a cat with refractory localised sporotrichosis. AB - The present report concerns a case of sporotrichosis in a 3-year-old male crossbred cat. After 9 months of oral itraconazole (20mg/kg) therapy, all skin lesions had resolved with the exception of a single nodular lesion located on the bridge of the nose. Therefore, a combined therapy that included intralesional (IL) amphotericin B (1mg/kg) was started. The lesion resolved completely after three weekly administrations of IL amphotericin B, given in concert with oral itraconazole. The cat remains well 1 year after discontinuing therapy, with no signs of recurrence. PMID- 19553148 TI - OPTESIM, a versatile toolbox for numerical simulation of electron spin echo envelope modulation (ESEEM) that features hybrid optimization and statistical assessment of parameters. AB - Electron spin echo envelope modulation (ESEEM) is a technique of pulsed-electron paramagnetic resonance (EPR) spectroscopy. The analyis of ESEEM data to extract information about the nuclear and electronic structure of a disordered (powder) paramagnetic system requires accurate and efficient numerical simulations. A single coupled nucleus of known nuclear g value (g(N)) and spin I=1 can have up to eight adjustable parameters in the nuclear part of the spin Hamiltonian. We have developed OPTESIM, an ESEEM simulation toolbox, for automated numerical simulation of powder two- and three-pulse one-dimensional ESEEM for arbitrary number (N) and type (I, g(N)) of coupled nuclei, and arbitrary mutual orientations of the hyperfine tensor principal axis systems for N>1. OPTESIM is based in the Matlab environment, and includes the following features: (1) a fast algorithm for translation of the spin Hamiltonian into simulated ESEEM, (2) different optimization methods that can be hybridized to achieve an efficient coarse-to-fine grained search of the parameter space and convergence to a global minimum, (3) statistical analysis of the simulation parameters, which allows the identification of simultaneous confidence regions at specific confidence levels. OPTESIM also includes a geometry-preserving spherical averaging algorithm as default for N>1, and global optimization over multiple experimental conditions, such as the dephasing time (tau) for three-pulse ESEEM, and external magnetic field values. Application examples for simulation of (14)N coupling (N=1, N=2) in biological and chemical model paramagnets are included. Automated, optimized simulations by using OPTESIM lead to a convergence on dramatically shorter time scales, relative to manual simulations. PMID- 19553151 TI - Feline exocrine pancreatic insufficiency: 16 cases (1992-2007). AB - Medical records of 16 cats diagnosed with exocrine pancreatic insufficiency (EPI) were reviewed. The diagnosis was confirmed with either a serum feline trypsin like immunoreactivity (fTLI) concentration 30 microg/l). At the end of the initial three months correction period, the plasma levels of 25(OH) D rose significantly. However, only 60% of patients reach a normal plasma concentration of calcidiol with the highest - 600,000UI - ergocalciferol cumulative dosage. At the end of the three months maintenance period, plasma 25(OH) D concentrations fell in all patients. No significant change was observed in parathormone, calcium, phosphates and 1-25(OH) D plasma levels. There was no hypercalcemic episode. CONCLUSION: KDOQI ergocalciferol recommended doses for stages 3 and 4 renal patients did not correct calcidiol deficiency in hemodialysis patients. New prospective studies are required for defining the modalities of an efficient vitamin D supplementation with ergocalciferol or cholecalciferol. PMID- 19553167 TI - [How can we integrate nutritional support in medical oncology?]. AB - Malnutrition concerns 20% of cancer patients. It is an independent risk factor for morbidity and mortality. And nutritional support has to be integrated in anti cancer treatment project, and could improve the prognosis and quality of life. Two axes in cancer cachexia care could be identified: nutritional and pharmacological. The first level of care includes dietetic counselling and oral nutritional supplements. When oral intakes are less 60% of needs, we have to use artificial nutrition, enteral if digestive tract is functional, otherwise parenteral nutrition. Immunonutrition in oncology concerns omega-3 fatty acids, some amino acids (arginine, glutamine, polyamins), the anti-oxidant micronutrients. Their indications and contraindications begin to emerge in oncology. Among molecules usefull in cancer cachexia, numerous were tested for their orexigen effect. The only ones, megestrol acetate and corticosteroids proved their efficacy in short cures. Today, nutrition is not any more palliative care but has to be included in anti-cancer treatment plan. According to the degree of malnutrition or malnutrition risk, different levels of nutritional support could be purposed. And immunonutrition could improve beneficial effects of nutritional support during cancer chemotherapy or radiotherapy. PMID- 19553168 TI - Antibacterial activity in vitro of Thymus capitatus from Jordan. AB - This study was carried out to evaluate the antibacterial activity of aqueous and organic extracts of Thymus capitatus L. (Lamiaceae) leaves and stems. Dried ground powder leaves and stems were extracted with water (aqueous extracts), ethanol, dichloromethane and hexane (Soxhlet extracts). The antibacterial activity of these extracts was evaluated against bacteria using disc diffusion method. The result obtained showed that the leaves had stronger antibacterial activity than the stems extracts. The ethanolic extract had the highest yield products and the high antibacterial activity than all other solvents. The results suggest that essential oil as non-polar organic compounds could be the main active compounds in this plant. Therefore the antibacterial activity of leaves ethanol extracts (LEE) was compared with essential oils leaves extracts (LEO) of T. capitatus. The LEO showed greater antibacterial activity than LEE. The LEO showed a broad spectrum of antibacterial activity and the Pseudomonas aeruginosa was the most sensitive bacteria. PMID- 19553170 TI - Diclofenac inhibits proliferation but not NGF-induced differentiation of PC12 cells. AB - Diclofenac is a non-steroidal anti-inflammatory drug that is prescribed for treatment of rheumatic diseases and as an analgesic. Although the information about these side effects has been widely reported, little is know about the effect of diclofenac on the neural cells. In this study, we investigated the effects of diclofenac on the proliferation and differentiation of PC12 cells. The cell proliferation was evaluated by using XTT assay in the both free-serum neurobasal medium supplemented with B27 supplement and DMEM/F12 medium containing 10% FBS. The nerve growth factor (NGF)-induced differentiation was assessed by measuring the neurite length. The drug toxicity was exhibited at the concentrations more than 310 microM in the supplemented neurobasal medium. The treatment of cells in the DMEM/F12 medium increased their sensitivity to diclofenac, with 40% and 75% growth inhibition at the 155 and 310 microM concentrations, respectively. The NGF-induced differentiation was not reduced by toxic and subtoxic concentrations of diclofenac. The results of this study indicated that diclofenac may be able to exhibit its neurotoxic effects through growth inhibition, but not differentiation inhibition. Supplement of B27 has several antioxidant compounds. Therefore, the difference of diclofenac cytotoxic effects in two culture media suggest that drug cytotoxicity may be related to the oxidative stress. PMID- 19553169 TI - Comparative study of two bacteriocins produced by representative indigenous soil bacteria. AB - The aim of this research work was to identify and characterized the bacteriocins produced by soil-associated bacteria. Bacillocin from Bacillus brevis Bb and pyocin from Pseudomonas aeruginosa Pa were found bioactive only against gram positive bacteria tested. Maximum production of both the bacteriocins was observed at 32 degrees C in BHI medium. Production of both the bacteriocins started in the early exponential growth phase while the maximum production was observed during the stationary phase. Bacillocin Bb remained stable during 1-9 pH while pyocin Pa remained stable at pH 1-11. Both of the bacterocins were found resistant to high temperature (100 degrees C for 30 min), detergents (1% solutions of EDTA, Tween 20, Tween 80 and SDS) and organic solvents (1% solutions of Ethanol, Butanol, Methanol, Propanol, Chloroform, and Acetone). Activity of both was completely lost after proteinase K treatment suggesting their protein nature. Titre of bacillocin Bb was estimated to be 5280 AU/mL while the titre for pyocin Pa was calculated as 640 AU/mL. Both of the bacteriocins showed bacteriolytic mode of action against the indicator Bacillus strain BC31 and were found <10 KDa in their molecular mass. PMID- 19553171 TI - Quantitation of alfuzosin hydrochloride in pharmaceutical formulations by RP HPLC. AB - An isocratic reversed phase high-performance liquid chromatographic (HPLC) method with ultraviolet detection at 245 nm has been developed for the determination of alfuzosin hydrochloride in dosage formulation. Good chromatographic separation alfuzosin was achieved by using a stainless steel analytical column Inertsil ODS 3V (5 microm, 15 cmx0.46 cm). The system was operated at ambient temperature (25+/-2 C) using a mobile phase consisting of acetonitrile:water:tetrahydrofuran:perchloricacid (250:740:10:1) at a flow rate of 1 ml/min. The calibration curve for alfuzosin hydrochloride was linear over the tested concentration range of 50%, 75%, 100%, 125% and 150% with reference to the label claim and a correlation coefficient of 0.999. The intra- and inter-run precision and accuracy results were 98.07 to 100.34 with the %RSD of 0.71% and tailings factor 1.07. The proposed method was validated for its selectivity, linearity, accuracy, and precision. The method was found to be suitable for the quality control of alfuzosin hydrochloride in bulk drug as well as in formulation. PMID- 19553172 TI - Comparison of hypotensive and hypolipidemic effects of Catharanthus roseus leaves extract with atenolol on adrenaline induced hypertensive rats. AB - The leaves extract of Catharanthus roseus was investigated for hypotensive and hypolipidemic effects in adrenaline-induced hypertensive rats (AIHR) and compared with those of Atenolol in a crossover design. The pharmacologically Active components responsible for hypotensive activities were isolated from plant using bioassay guided purification approach and the structure of the compounds was proposed by spectroscopic methods. Catharanthus roseus leaves extract and commercial drug Atenolol were administered through intraperitoneal (i.p) route for one week. Different biochemical parameters such as heart weight, blood glucose level, serum cholesterol level, serum triglyceride level, body weight and the relationships between them were measured. Catharanthus roseus leaves extract at a dose of 30 mg/155+/-15 gm of body weight was injected in rat at every morning during the treatment period. The dose of Atenolol was determined according to its pharmacokinetic parameters. Clinically effective plasma concentration as a hypotensive drug was obtained after the injection of 0.1 mg/155+/-15 gm of body weight of the drug. The Catharanthus roseus leaves extract made significant changes in each cardiovascular parameter after investigation. Catharanthus roseus leaves extract treated animals have shown the hypotensive effects. Hypotensive effects were also shown by Atenolol. PMID- 19553173 TI - Effect of aqueous extracts of alligator pear seed (Persea americana mill) on blood glucose and histopathology of pancreas in alloxan-induced diabetic rats. AB - Effects of aqueous extract of alligator pear seed on normal and alloxan-induced diabetic rats were investigated in 6 groups of rats (5 rats per group). Test groups were made diabetic with intra-peritoneal injection of alloxan and treated with 300 mg and 600 mg/kg body weight of alligator pear seed extract. Two non diabetic groups were also administered with 300 mg and 600 mg/kg body weight extract. The levels of blood glucose were examined in all 6 experimental groups. In diabetic rats, blood glucose levels were significantly reduced (p<0.05) by 73.26-78.24% on consumption of the extracts, with greater effect exhibited by the 600 mg/kg extract. In normal rats, blood glucose levels were significantly reduced (p<0.05) by 34.68-38.9% on consumption of the seed extract. Histological studies showed a degenerative effect on the pancreatic islet cells of diabetic rats. The result suggested restorative (protective) effect of the extract on pancreatic islet cells. Administration of aqueous extract of alligator pear seed may contribute significantly to the reduction of blood glucose levels and can be useful in the treatment of diabetes. PMID- 19553174 TI - Antioxidant activity, phenol and flavonoid contents of 13 citrus species peels and tissues. AB - Methanolic extracts of 13 commercially available citrus spp., peels and tissues growing in Iran were investigated for their antioxidant activity by DPPH method. IC50 for antioxidant activity ranged from 0.6-3.8 mg ml(-1). Total phenolic content of the citrus spp. samples (based on folin Ciocalteu method) varied from 66.5 to 396.8 mg gallic acid equivalent/g of extract and flavonoids content (based on colorimetric AlCl3 method) varied from 0.3 to 31.1 mg quercetin equivalent/g of extract. There were no correlation between the total phenolic and/or flavonoids contents and antioxidant activity in tissues and/or peels. PMID- 19553175 TI - Evaluation of antibacterial activity, phenol and flavonoid contents of Thespesia populnea flower extracts. AB - Flavonoids which were reported as having many pharmacological activities, antimicrobial, antioxidant, cytotoxic, chemoprevention activities and they possess strong antiproliferative effects related to inhibition of cell cycle progression and apoptosis induction. On the basis of this Thespesia populnea (L.) Sol. Ex Correa (Family-Malvaceae) was selected and it is having the major composition of flavonoids and the antibacterial activity of methanolic extract of Thespesia populnea flowers was investigated by agar well diffusion method. Furthermore our phytochemical studies indicated that methanolic extract of Thespesia populnea flowers contains flavonoids, alkaloids, tannins and anthroquinone glycosides. Moreover the individual components were identified by thin layer chromatography and Rf value was compared with standard flavonoid quercetin. The total phenolic and flavonoid content studies were also quantified. The bacteria used for antibacterial study were Shigella flexneri (NCIM 4924), Rhodococcus terrae (NCIM 5126), Escherichiae coli (ATCC 11775), Streptococcus faecalis (NCIB 2406), Klebsiella pneumoniae (ATCC 13883),Brevibacterium luteum (NCIM 2923), Micrococcus flavum (NCIM 2376), Proteus mirabilis (NCIB 8268), Bacillus licheniformis (NCIM 2468), Micrococcus luteus (ATCC 2984), Flavobacterium devorans (NCIM 2581), Shigella sonei(ATCC 29930), Shigella boydii (ATCC 8700) and Shigella dysentriae (ATCC 13313).According to our results in the lowest tested concentration of 62.5 microg/ml and 125mug/ml 7.2% of the plant extract were active, 5% active in the concentration of 250 microg/ml, 75.7% active in the concentration of 500 microg/ml and 92.8% active at the concentration of 1000 microg/ml in a dose dependent manner. PMID- 19553176 TI - Antinociceptive and anti-pyretic activity of Benincasa hispida (thunb.) cogn. in Wistar albino rats. AB - The seeds of Benincasa hispida (Thunb) COGN. (Family: Cucurbitaceae) was extracted with ethanol and was used to study acute toxicity, antinociceptive and anti-pyretic effects. Brewer's yeast (15%) was used to induce pyrexia in rats. The extract was non lethal to the rats up to the dose of 5000 mg/kg b.w. At doses of 250 and 500 mg/kg b.w, the extract significantly (P<0.05) increased the antinociceptive effective in a dose dependent manner in rats. Similarly, at doses of 250 and 500 mg/kg b.w the extract significantly (P<0.05) decreased yeast induced pyrexia in rats. These results indicate that ethanolic extract of Benincasa hispida possesses potent antinociceptive and antipyretic effects and thus pharmacologically justifying its folkloric use in the management of fever and pain conditions. PMID- 19553177 TI - A comparative study of various microencapsulation techniques: effect of polymer viscosity on microcapsule characteristics. AB - It is a comparative study of salbutamol sulphate-ethylcellulose microcapsules prepared by three different microencapsulation techniques i.e. coacervation thermal change, solvent evaporation and coacervation non-solvent addition by adjusting the ratio of salbutamol sulphate to ethylcellulose. In vitro release profiles of microcapsules were studied using USP XXIV dissolution apparatus-I in 450 ml double distilled water maintained at 37 degrees C at 50 rpm. Scanning electron microscopic results indicated that all microcapsules were aggregated, whitish and irregular in shape with good entrapment efficiency (86.34 to 97.83), production yield (87.91+/-1.34 to 98.33+/-1.37) and flow properties. Initial burst effect was observed in the drug release behavior from all microcapsules. A slight increase in actual drug loading but profound increase in mean diameter of microcapsules was observed with the increase in the viscosity of ethylcellulose. UV and FTIR spectroscopy, x-ray diffractometry and thermal analysis verified the absence of any strong chemical interaction between drugs and polymer. The drug release from all the formulations followed anomalous diffusion mechanism and was best fit to Higuchi's kinetic model. The results suggest coacervation thermal change as an appropriate approach to develop slow-release multi-unit oral dosage form of salbutamol sulphate suggesting at least twice administration in every 24 hours. PMID- 19553178 TI - Quantitative determination of levofloxacin hemihydrate in bulk and tablets by UV spectrophotometry and first order derivative methods. AB - Two simple, rapid, accurate and economical 'UV Spectrophotometry' and 'First Order Derivative' methods have been developed for determination of levofloxacin hemihydrate in bulk and tablets. In (10% v/v) acetonitrile, the lambdamax of the drug was found to be 288 nm. The same spectrum was derivatised into first order derivative, using UV probe software of instrument (Shimadzu-2450), at Deltalambda=4. The amplitude of the trough was recorded at 297 nm. In both the proposed methods, levofloxacin hemihydrate follows linearity in the concentration range 2-12 microg/ml with a correlation coefficient of 0.9999. Assay results were in good agreement with label claim. The methods were validated statistically and by recovery studies. The relative standard deviation were found to be less than 2% with excellent precision and accuracy. PMID- 19553179 TI - Effect of channeling agents on the release pattern of theophylline from kollidon SR based matrix tablets. AB - The purpose of the present study was to investigate the effect of channeling agent on the release profile of theophylline from Kollidon SR based matrix systems. Matrix tablets of theophylline using Kollidon SR which is plastic in nature were prepared by direct compression process. NaCl and PEG 1500 were used as channeling agents. Drug release study was evaluated for eight hours using USP 22 paddle-type dissolution apparatus using distilled water as the dissolution medium. The release mechanisms were explored and explained with zero order, Higuchi, first order and Korsmeyer equations. The release rate, extent and mechanisms were found to be governed by the type and content of the channeling agents. Increased rate and extent of the drug release were found by using higher content of channeling agent (42.49%) in the matrix due to increased porosity when compared with the formulation having no channeling agents. On the other hand decreased rate and extent of drug release were observed in the formulation having lower channeling agent content (19.76%). PEG 1500 ensures maximum release of drug from Kollidon SR than NaCl when other parameters were kept unchanged. It was found that type and amount of channeling agent significantly affect the time required for 50% of drug release (T50%), percentage drug release at 8 hours, release rate constant (K) and diffusion exponent (n). Kinetic modeling of dissolution profiles revealed drug release mechanism ranges from diffusion controlled or Fickian transport to anomalous type or non-Fickian transport, which was mainly dependent on the type and amount of channeling agents. These studies indicate that the proper balance between a matrix forming agent and a channeling agent can produce a drug dissolution profile similar to a desired dissolution profile. PMID- 19553180 TI - Effect of methanolic extract of Hyoscymus niger L. on the seizure induced by picritoxin in mice. AB - Effects of Hyoscyamus niger L on central nervous system have been known for many years. The effects of methanolic extract of H. niger L. on seizures induced by picrotoxin was studied in mice in this investigation. In this study 7 groups of animals pretreated with methanolic extract of the plant (12.5, 25, 50, 100, 200, 300, 400 mg/kg/i.p.), 20 minutes prior to the picrotoxin (12 mg/kg/ i.p.)- induced seizures. Control mice received phenobarbital (40 mg/kg/i.p.) as positive control, or saline (10 ml/kg) as negative control. The latency of seizure (sec), duration of seizure (sec) and mortality rate were determined in test and control groups. The results of this study showed that latency of seizure was increased in groups that were pretreated with doses of 100, 200, 300 and 400 mg/kg of extract. In addition, methanolic extract of H. niger L. delayed the death time in mice as compared to control that was significant with doses of 200, 300 and 400 mg/kg. The most effective dose of extract was 300 mg/kg in this investigation (P<0.01). In conclusion, the results showed that methanolic extract of H. niger L. posses the anticonvulsant activity against picrotoxin-induced seizures in mice. The exact mechanism(s) by which the plant exerts its anticonvulsant activity is not determined yet. PMID- 19553181 TI - Caffeic acid phenethyl ester decreases the level of S-100B protein after middle cerebral artery [correction for after] occlusion in rabbits. AB - Effects of caffeic acid phenethyl ester (CAPE) on the serum S-100B levels were studied as an index for brain damage after permanent middle cerebral artery (MCA) occlusion in rabbits. Twenty rabbits were divided into four groups (n=5): control, sham, non-treatment and CAPE. The right MCA was occluded using a microsurgical procedure with bipolar coagulation and was then transected in non treatment and CAPE groups. The rabbits in the sham group underwent a surgical procedure but the MCA was not occluded. No surgery was performed in the control group. CAPE was administered after MCA occlusion at the dose of 10 microg/kg, once a day intraperitoneally for 7 days in the CAPE group. Serum S-100B levels were determined on days 1, 2, 4 and 7. Serum S-100B level was significantly increased following permanent MCA occlusion. Posttreatment of CAPE significantly reduced the serum S-100B level. This study demonstrated that CAPE is capable of attenuating increased serum S-100B level induced by MCA occlusion in rabbits. CAPE may be useful as a neuroprotective agent. PMID- 19553182 TI - Bioassay screening of the essential oil and various extracts from 4 spices medicinal plants. AB - Four commonly used spices plants in Iran were evaluated for cytotoxicity effect using Brine Shrimp Lethality (BSL) assay. Essential oils and various extracts of Heracleum persicum, Nigella arvensis, Cinnamomum zeylanicum and Zingiber officinale were assessed by two methods of disk and solution of BSL. Data were processed in probit-analysis program to estimate LC50 values. All of the tested fractions have exhibited more cytotoxicity in the solution method. Essential oils of H. persicum and C. zeylanicum have shown the most cytotoxicity with LC50 values 0.007 and 0.03 microg/ml respectively. None of aqueous extracts showed significant cytotoxicity. The analysis of the essential oil of H. persicum showed the hexyl butyrate and octyl acetate as the main compounds. These results suggest some limitation for using of these spices in diet. Furthermore, these plants could be considered as a source of cytotoxic compounds which might be studied in more details. PMID- 19553183 TI - Simultaneous determination of metronidazole and miconazole nitrate in gel by HPTLC. AB - A new, simple, precise, rapid and selective high-performance thin-layer chromatographic (HPTLC) method for the simultaneous quantification of Metronidazole (MTZ) and Miconazole nitrate (MCZ) in gel has been developed. It was performed on silica gel 60 GF254 Thin Layer Chromatographic plates using mobile phase comprising of Toluene: Chloroform: Methanol (3.0:2.0:0.6 v/v) and the detection was carried out at 240 nm using densitometer. The retention factors of MTZ and MCZ were 0.34 and 0.55 respectively. Calibration curves were linear in the range of 300-700 ng/spot of MTZ and 600-1400 ng/spot of MCZ both by height and by area. The percent recovery of the drugs from gel carried out by standard addition method was found to be 100.13+/-1.59 (by height) and 98.92+/-0.76 (by area) for MTZ and 99.49+/-1.58 (by height) and 99.63+/-1.46 (by area) for MCZ indicative of accuracy and precision of simultaneous determination of MTZ and MCZ nitrate. PMID- 19553184 TI - Bioavailability of ciprofloxacin tablets in humans and its correlation with the dissolution rates. AB - Bioavailabilities of seven tablets of ciprofloxacin were determined. The correlation between in-vivo bioavailability parameters and in-vitro dissolution rates were studied. Ciprofloxacin concentration from the blood was determined by microbiological assay technique. The release pattern of ciprofloxacin from tablets, which were determined by dissolution USP paddle method and spectrophotometric method was used to determine the concentration. All the parameters of ciprofloxacin bioavailability i.e., Tmax, Cmax, AUC and absorption rate constant (Ka) showed no significant correlation with dissolution rates at T30%, T50%, T90% and dissolution at 30 mins with aims as in-vivo bioequvalence waiver. The value of dissolution test used as quality tool for predicting in-vivo performance of drug product is significantly enhanced, if the in-vivo-in-vitro relationship (correlation or association) is established. PMID- 19553185 TI - Report: In vitro antimicrobial activities of extracts of Carpolobia lutea root. AB - Several plants are used in folk medicine to treat infections. Carpolobia lutea, G.Don (Polygalaceae) is a medicinal plant commonly used by herbalists in Southern Nigeria against dental and genitourinary infections. The study was to evaluate the in vitro antimicrobial activities of n-hexane, chloroform, ethyl acetate and methanol extracts of Carpolobia lutea root. Four typed cultures of bacteria namely, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and two clinical strains of fungi, namely Candida albicans and Tinea capitis were analyzed using agar well diffusion method. The extracts that showed antimicrobial activity were then tested to determine the Minimum Inhibitory Concentration for each bacterial or fungal sample.The ethyl acetate extract gave the widest zone of inhibition (21.0 mm) followed by chloroform when tested on E. coli. No inhibition was observed with E.Coli. None of the extracts showed any inhibitory effect against Pseudomonas aeruginosa and the fungal strains of Candida albicans and Tinea capitis. The most potent of these extracts was Chloroform extract with Minimum Inhibitory Concentration (MIC) of 25 mg/ml for bacteria. The phytochemical screening of the root of C. lutea revealed the presence of saponins, anthraquinones, flavonoids, cardiac glycosides, simple sugar and terpenes. PMID- 19553186 TI - Report: antibiotic production by thermophilic Bacillus specie SAT-4. AB - Production of antimicrobial compounds seems to be a general phenomenon for most bacteria. The prevalence of antimicrobial resistance among key microbial pathogens is increasing at an alarming rate worldwide. Current solutions involve development of a more rationale approach to antibiotic use and discover of new antimicrobials. Bacillus species produce a large number of biological compounds active against bacteria, fungi, protozoa and viruses. The process of production usually involves screening of wide range of microorganisms, testing and modification. Production is carried out using fermentation. Thermophilic spore forming, gram positive, motile rod bacterial strains were isolated from the Thar Desserts, Sindh Province, Pakistan. These strains were screened and checked for antibacterial activity. The best activity was observed by SAT4 against Micrococcus luteus, Staphylococcus aureus and Pseudomonas aeroginosa. The activity was only observed against gram positive bacteria and no activity was seen against Pseudomonas aeroginosa. Thermophilic Bacillus specie SAT4 was found to be active in the fermentation process to produce the antimicrobial agents. Further optimizations of different conditions (time of incubation, media, pH, glucose concentrations, nitrogen concentrations, and temperature) for antimicrobial production by the selected bacterial strain was performed. Agar diffusion assay was performed to evaluate the antibacterial activity. Optimum conditions for the production of antimicrobials by selected isolate were observed to be 48 hour, pH 5, temperature 55 degrees C, 2% glucose and 1.5% nitrogen concentration. This newly isolated bacterial strain has great potential for antimicrobial production at industrial scale. PMID- 19553187 TI - The role of pomegranate (Punica granatum L.) in colon cancer. AB - Colon cancer is one of the major causes of cancer-related death in the Western world. Although cytotoxic chemotherapeutic agents are available to treat the disease, these agents become ineffective as the disease advances to an invasive state. An alternative but viable approach to reduce the incidence of this deadly disease is then, to increase the dietary intake of relatively non-toxic fruits and vegetables. An example of a fruit with antioxidant, antidiabetic and anti atherosclerotic properties is pomegranate. Pomegranate produces anticancer effects in experimental models of lung, prostate and skin cancer. More recently, pomegranate has been found to be anti-carcinogenic in the colon. This communication discusses pomegranate's effect in colon cancer. PMID- 19553188 TI - Analysis of repair mechanism choice during homologous recombination. AB - Double-strand breaks (DSBs) occur frequently during cell growth. Due to the presence of repeated sequences in the genome, repair of a single DSB can result in gene conversion, translocation, deletion or tandem duplication depending on the mechanism and the sequence chosen as partner for the recombinational repair. Here, we study how yeast cells repair a single, inducible DSB when there are several potential donors to choose from, in the same chromosome and elsewhere in the genome. We systematically investigate the parameters that affect the choice of mechanism, as well as its genetic regulation. Our results indicate that intrachromosomal homologous sequences are always preferred as donors for repair. We demonstrate the occurrence of a novel tri-partite repair product that combines ectopic gene conversion and deletion. In addition, we show that increasing the distance between two repeated sequences enhances the dependence on Rad51 for colony formation after DSB repair. This is due to a role of Rad51 in the recovery from the checkpoint signal induced by the DSB. We suggest a model for the competition between the different homologous recombination pathways. Our model explains how different repair mechanisms are able to compensate for each other during DSB repair. PMID- 19553189 TI - Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis. AB - Double-stranded (ds), as well as denatured, single-stranded (ss) DNA samples can be analyzed on urea-agarose gels. Here we report that after denaturation by heat in the presence of 8 M urea, the two strands of the same ds DNA fragment of approximately 1-20-kb size migrate differently in 1 M urea containing agarose gels. The two strands are readily distinguished on Southern blots by ss-specific probes. The different migration of the two strands could be attributed to their different, base composition-dependent conformation impinging on the electrophoretic mobility of the ss molecules. This phenomenon can be exploited for the efficient preparation of strand-specific probes and for the separation of the complementary DNA strands for subsequent analysis, offering a new tool for various cell biological research areas. PMID- 19553190 TI - A trans-splicing group I intron and tRNA-hyperediting in the mitochondrial genome of the lycophyte Isoetes engelmannii. AB - Plant mitochondrial genomes show much more evolutionary plasticity than those of animals. We analysed the first mitochondrial DNA (mtDNA) of a lycophyte, the quillwort Isoetes engelmannii, which is separated from seed plants by more than 350 million years of evolution. The Isoetes mtDNA is particularly rich in recombination events, and chloroplast as well as nuclear DNA inserts document the incorporation of foreign sequences already in this most ancestral vascular plant lineage. On the other hand, particularly small group II introns and short intergenic regions reveal a tendency of evolution towards a compact mitochondrial genome. RNA editing reaches extreme levels exceeding 100 pyrimidine exchanges in individual mRNAs and, hitherto unobserved in such frequency, also in tRNAs with 18 C-to-U conversions in the tRNA for proline. In total, some 1500 sites of RNA editing can be expected for the Isoetes mitochondrial transcriptome. As a unique molecular novelty, the Isoetes cox1 gene requires trans-splicing via a discontinuous group I intron demonstrating disrupted, but functional, RNAs for yet another class of natural ribozymes. PMID- 19553191 TI - BRCA2-dependent homologous recombination is required for repair of Arsenite induced replication lesions in mammalian cells. AB - Arsenic exposure constitutes one of the most widespread environmental carcinogens, and is associated with increased risk of many different types of cancers. Here we report that arsenite (As[III]) can induce both replication dependent DNA double-strand breaks (DSB) and homologous recombination (HR) at doses as low as 5 microM (0.65 mg/l), which are within the typical doses often found in drinking water in contaminated areas. We show that the production of DSBs is dependent on active replication and is likely to be the result of conversion of a DNA single-strand break (SSB) into a toxic DSB when encountered by a replication fork. We demonstrate that HR is required for the repair of these breaks and show that a functional HR pathway protects against As[III]-induced cytotoxicity. In addition, BRCA2-deficient cells are sensitive to As[III] and we suggest that As[III] could be exploited as a therapy for HR-deficient tumours such as BRCA1 and BRCA2 mutated breast and ovarian cancers. PMID- 19553192 TI - Comparative analysis of activator-Esigma54 complexes formed with nucleotide-metal fluoride analogues. AB - Bacterial RNA polymerase (RNAP) containing the major variant sigma(54) factor forms open promoter complexes in a reaction in which specialized activator proteins hydrolyse ATP. Here we probe binding interactions between sigma(54)-RNAP (Esigma(54)) and the ATPases associated with various cellular activities (AAA+) domain of the Escherichia coli activator protein, PspF, using nucleotide-metal fluoride (BeF and AlF) analogues representing ground and transition states of ATP, which allow complexes (that are otherwise too transient with ATP) to be captured. We show that the organization and functionality of the ADP-BeF- and ADP AlF-dependent complexes greatly overlap. Our data support an activation pathway in which the initial ATP-dependent binding of the activator to the Esigma(54) closed complex results in the re-organization of Esigma(54) with respect to the transcription start-site. However, the nucleotide-dependent binding interactions between the activator and the Esigma(54) closed complex are in themselves insufficient for forming open promoter complexes when linear double-stranded DNA is present in the initial closed complex. PMID- 19553193 TI - Preparation of selective and segmentally labeled single-stranded DNA for NMR by self-primed PCR and asymmetrical endonuclease double digestion. AB - We demonstrate a new, efficient and easy-to-use method for enzymatic synthesis of (stereo-)specific and segmental (13)C/(15)N/(2)H isotope-labeled single-stranded DNA in amounts sufficient for NMR, based on the highly efficient self-primed PCR. To achieve this, new approaches are introduced and combined. (i) Asymmetric endonuclease double digestion of tandem-repeated PCR product. (ii) T4 DNA ligase mediated ligation of two ssDNA segments. (iii) In vitro dNTP synthesis, consisting of in vitro rNTP synthesis followed by enzymatic stereo-selective reduction of the C2' of the rNTP, and a one-pot add-up synthesis of dTTP from dUTP. The method is demonstrated on two ssDNAs: (i) a 36-nt three-way junction, selectively (13)C(9)/(15)N(3)/(2)H((1',2'',3',4',5',5''))-dC labeled and (ii) a 39-nt triple-repeat three-way junction, selectively (13)C(9)/(15)N(3)/(2)H((1',2'',3',4',5',5''))-dC and (13)C(9)/(15)N(2)/(2)H((1',2'',3',4',5',5''))-dT labeled in segment C20-C39. Their NMR spectra show the spectral simplification, while the stereo-selective (2)H-labeling in the deoxyribose of the dC-residues, straightforwardly provided assignment of their C1'-H2' and C2'-H2' resonances. The labeling protocols can be extended to larger ssDNA molecules and to more than two segments. PMID- 19553194 TI - Structural basis for the sequence-specific RNA-recognition mechanism of human CUG BP1 RRM3. AB - The CUG-binding protein 1 (CUG-BP1) is a member of the CUG-BP1 and ETR-like factors (CELF) family or the Bruno-like family and is involved in the control of splicing, translation and mRNA degradation. Several target RNA sequences of CUG BP1 have been predicted, such as the CUG triplet repeat, the GU-rich sequences and the AU-rich element of nuclear pre-mRNAs and/or cytoplasmic mRNA. CUG-BP1 has three RNA-recognition motifs (RRMs), among which the third RRM (RRM3) can bind to the target RNAs on its own. In this study, we solved the solution structure of the CUG-BP1 RRM3 by hetero-nuclear NMR spectroscopy. The CUG-BP1 RRM3 exhibited a noncanonical RRM fold, with the four-stranded beta-sheet surface tightly associated with the N-terminal extension. Furthermore, we determined the solution structure of the CUG-BP1 RRM3 in the complex with (UG)(3) RNA, and discovered that the UGU trinucleotide is specifically recognized through extensive stacking interactions and hydrogen bonds within the pocket formed by the beta-sheet surface and the N-terminal extension. This study revealed the unique mechanism that enables the CUG-BP1 RRM3 to discriminate the short RNA segment from other sequences, thus providing the molecular basis for the comprehension of the role of the RRM3s in the CELF/Bruno-like family. PMID- 19553195 TI - Extracting transcription factor targets from ChIP-Seq data. AB - ChIP-Seq technology, which combines chromatin immunoprecipitation (ChIP) with massively parallel sequencing, is rapidly replacing ChIP-on-chip for the genome wide identification of transcription factor binding events. Identifying bound regions from the large number of sequence tags produced by ChIP-Seq is a challenging task. Here, we present GLITR (GLobal Identifier of Target Regions), which accurately identifies enriched regions in target data by calculating a fold change based on random samples of control (input chromatin) data. GLITR uses a classification method to identify regions in ChIP data that have a peak height and fold-change which do not resemble regions in an input sample. We compare GLITR to several recent methods and show that GLITR has improved sensitivity for identifying bound regions closely matching the consensus sequence of a given transcription factor, and can detect bona fide transcription factor targets missed by other programs. We also use GLITR to address the issue of sequencing depth, and show that sequencing biological replicates identifies far more binding regions than re-sequencing the same sample. PMID- 19553196 TI - Interplay of DNA supercoiling and catenation during the segregation of sister duplexes. AB - The discrete regulation of supercoiling, catenation and knotting by DNA topoisomerases is well documented both in vivo and in vitro, but the interplay between them is still poorly understood. Here we studied DNA catenanes of bacterial plasmids arising as a result of DNA replication in Escherichia coli cells whose topoisomerase IV activity was inhibited. We combined high-resolution two-dimensional agarose gel electrophoresis with numerical simulations in order to better understand the relationship between the negative supercoiling of DNA generated by DNA gyrase and the DNA interlinking resulting from replication of circular DNA molecules. We showed that in those replication intermediates formed in vivo, catenation and negative supercoiling compete with each other. In interlinked molecules with high catenation numbers negative supercoiling is greatly limited. However, when interlinking decreases, as required for the segregation of newly replicated sister duplexes, their negative supercoiling increases. This observation indicates that negative supercoiling plays an active role during progressive decatenation of newly replicated DNA molecules in vivo. PMID- 19553197 TI - Degradation of nanoRNA is performed by multiple redundant RNases in Bacillus subtilis. AB - Escherichia coli possesses only one essential oligoribonuclease (Orn), an enzyme that can degrade oligoribonucleotides of five residues and shorter in length (nanoRNA). Firmicutes including Bacillus subtilis do not have an Orn homolog. We had previously identified YtqI (NrnA) as functional analog of Orn in B. subtilis. Screening a genomic library from B. subtilis for genes that can complement a conditional orn mutant, we identify here YngD (NrnB) as a second nanoRNase in B. subtilis. Like NrnA, NrnB is a member of the DHH/DHHA1 protein family of phosphoesterases. NrnB degrades nanoRNA 5-mers in vitro similarily to Orn. Low expression levels of NrnB are sufficient for orn complementation. YhaM, a known RNase present in B. subtilis, degrades nanoRNA efficiently in vitro but requires high levels of expression for only partial complementation of the orn(-) strain. A triple mutant (nrnA(-), nrnB(-), yhaM(-)) in B. subtilis is viable and shows almost no impairment in growth. Lastly, RNase J1 seems also to have some 5'-to-3' exoribonuclease activity on nanoRNA and thus can potentially finish degradation of RNA. We conclude that, unlike in E. coli, degradation of nanoRNA is performed in a redundant fashion in B. subtilis. PMID- 19553198 TI - International guidelines for the diagnosis and management of hereditary haemorrhagic telangiectasia. AB - BACKGROUND: HHT is an autosomal dominant disease with an estimated prevalence of at least 1/5000 which can frequently be complicated by the presence of clinically significant arteriovenous malformations in the brain, lung, gastrointestinal tract and liver. HHT is under-diagnosed and families may be unaware of the available screening and treatment, leading to unnecessary stroke and life threatening hemorrhage in children and adults. OBJECTIVE: The goal of this international HHT guidelines process was to develop evidence-informed consensus guidelines regarding the diagnosis of HHT and the prevention of HHT-related complications and treatment of symptomatic disease. METHODS: The overall guidelines process was developed using the AGREE framework, using a systematic search strategy and literature retrieval with incorporation of expert evidence in a structured consensus process where published literature was lacking. The Guidelines Working Group included experts (clinical and genetic) from eleven countries, in all aspects of HHT, guidelines methodologists, health care workers, health care administrators, HHT clinic staff, medical trainees, patient advocacy representatives and patients with HHT. The Working Group determined clinically relevant questions during the pre-conference process. The literature search was conducted using the OVID MEDLINE database, from 1966 to October 2006. The Working Group subsequently convened at the Guidelines Conference to partake in a structured consensus process using the evidence tables generated from the systematic searches. RESULTS: The outcome of the conference was the generation of 33 recommendations for the diagnosis and management of HHT, with at least 80% agreement amongst the expert panel for 30 of the 33 recommendations. PMID- 19553199 TI - Re-evaluation of forest biomass carbon stocks and lessons from the world's most carbon-dense forests. AB - From analysis of published global site biomass data (n = 136) from primary forests, we discovered (i) the world's highest known total biomass carbon density (living plus dead) of 1,867 tonnes carbon per ha (average value from 13 sites) occurs in Australian temperate moist Eucalyptus regnans forests, and (ii) average values of the global site biomass data were higher for sampled temperate moist forests (n = 44) than for sampled tropical (n = 36) and boreal (n = 52) forests (n is number of sites per forest biome). Spatially averaged Intergovernmental Panel on Climate Change biome default values are lower than our average site values for temperate moist forests, because the temperate biome contains a diversity of forest ecosystem types that support a range of mature carbon stocks or have a long land-use history with reduced carbon stocks. We describe a framework for identifying forests important for carbon storage based on the factors that account for high biomass carbon densities, including (i) relatively cool temperatures and moderately high precipitation producing rates of fast growth but slow decomposition, and (ii) older forests that are often multiaged and multilayered and have experienced minimal human disturbance. Our results are relevant to negotiations under the United Nations Framework Convention on Climate Change regarding forest conservation, management, and restoration. Conserving forests with large stocks of biomass from deforestation and degradation avoids significant carbon emissions to the atmosphere, irrespective of the source country, and should be among allowable mitigation activities. Similarly, management that allows restoration of a forest's carbon sequestration potential also should be recognized. PMID- 19553201 TI - Binding-site geometry and flexibility in DC-SIGN demonstrated with surface force measurements. AB - The dendritic cell receptor DC-SIGN mediates pathogen recognition by binding to glycans characteristic of pathogen surfaces, including those found on HIV. Clustering of carbohydrate-binding sites in the receptor tetramer is believed to be critical for targeting of pathogen glycans, but the arrangement of these sites remains poorly understood. Surface force measurements between apposed lipid bilayers displaying the extracellular domain of DC-SIGN and a neoglycolipid bearing an oligosaccharide ligand provide evidence that the receptor is in an extended conformation and that glycan docking is associated with a conformational change that repositions the carbohydrate-recognition domains during ligand binding. The results further show that the lateral mobility of membrane-bound ligands enhances the engagement of multiple carbohydrate-recognition domains in the receptor oligomer with appropriately spaced ligands. These studies highlight differences between pathogen targeting by DC-SIGN and receptors in which binding sites at fixed spacing bind to simple molecular patterns. PMID- 19553200 TI - An essential role for ATP binding and hydrolysis in the chaperone activity of GRP94 in cells. AB - Glucose-regulated protein 94 (GRP94) is an endoplasmic reticulum (ER) chaperone for which only few client proteins and no cofactors are known and whose mode of action is unclear. To decipher the mode of GRP94 action in vivo, we exploited our finding that GRP94 is necessary for the production of insulin-like growth factor (IGF)-II and developed a cell-based functional assay. Grp94(-/-) cells are hypersensitive to serum withdrawal and die. This phenotype can be complemented either with exogenous IGF-II or by expression of functional GRP94. Fusion proteins of GRP94 with monomeric GFP (mGFP) or mCherry also rescue the viability of transiently transfected, GRP94-deficient cells, demonstrating that the fusion proteins are functional. Because these constructs enable direct visualization of chaperone-expressing cells, we used this survival assay to assess the activities of GRP94 mutants that are defective in specific biochemical functions in vitro. Mutations that abolish binding of adenosine nucleotides cannot support growth in serum-free medium. Similarly, mutations of residues needed for ATP hydrolysis also render GRP94 partially or completely nonfunctional. In contrast, an N terminal domain mutant that cannot bind peptides still supports cell survival. Thus the peptide binding activity in vitro can be uncoupled from the chaperone activity toward IGF in vivo. This mutational analysis suggests that the ATPase activity of GRP94 is essential for chaperone activity in vivo and that the essential protein-binding domain of GRP94 is distinct from the N-terminal domain. PMID- 19553202 TI - Prochlorococcus: approved for export. PMID- 19553205 TI - Locating the transition from periodic oscillations to spatiotemporal chaos in the wake of invasion. AB - In systems with cyclic dynamics, invasions often generate periodic spatiotemporal oscillations, which undergo a subsequent transition to chaos. The periodic oscillations have the form of a wavetrain and occur in a band of constant width. In applications, a key question is whether one expects spatiotemporal data to be dominated by regular or irregular oscillations or to involve a significant proportion of both. This depends on the width of the wavetrain band. Here, we present mathematical theory that enables the direct calculation of this width. Our method synthesizes recent developments in stability theory and computation. It is developed for only 1 equation system, but because this is a normal form close to a Hopf bifurcation, the results can be applied directly to a wide range of models. We illustrate this by considering a classic example from ecology: wavetrains in the wake of the invasion of a prey population by predators. PMID- 19553203 TI - APC/C-CCS52A complexes control meristem maintenance in the Arabidopsis root. AB - Plant organs originate from meristems where stem cells are maintained to produce continuously daughter cells that are the source of different cell types. The cell cycle switch gene CCS52A, a substrate specific activator of the anaphase promoting complex/cyclosome (APC/C), controls the mitotic arrest and the transition of mitotic cycles to endoreduplication (ER) cycles as part of cell differentiation. Arabidopsis, unlike other organisms, contains 2 CCS52A isoforms. Here, we show that both of them are active and regulate meristem maintenance in the root tip, although through different mechanisms. The CCS52A1 activity in the elongation zone of the root stimulates ER and mitotic exit, and contributes to the border delineation between dividing and expanding cells. In contrast, CCS52A2 acts directly in the distal region of the root meristem to control identity of the quiescent center (QC) cells and stem cell maintenance. Cell proliferation assays in roots suggest that this control involves CCS52A2 mediated repression of mitotic activity in the QC cells. The data indicate that the CCS52A genes favor a low mitotic state in different cell types of the root tip that is required for meristem maintenance, and reveal a previously undescribed mechanism for APC/C mediated control in plant development. PMID- 19553204 TI - Global distribution of conformational states derived from redundant models in the PDB points to non-uniqueness of the protein structure. AB - It is commonly accepted that proteins have evolutionarily conserved 3-dimensional structures, uniquely defined by their amino acid sequence. Here, we question the direct association of structure to sequence by comparing multiple models of identical proteins. Rapidly growing structural databases contain models of proteins determined independently multiple times. We have collected these models in the database of the redundant sets of protein structures and then derived their conformational states by clustering the models with low root-mean-square deviations (RMSDs). The distribution of conformational states represented in these sets is wider than commonly believed, in fact exceeding the possible range of structure determination errors, by at least an order of magnitude. We argue that differences among the models represent the natural distribution of conformational states. Our results suggest that we should change the common notion of a protein structure by augmenting a single 3-dimensional model by the width of the ensemble distribution. This width must become an indispensible attribute of the protein description. We show that every protein contains regions of high rigidity (solid-like) and regions of high mobility (liquid-like) in different and characteristic contribution. We also show that the extent of local flexibility is correlated with the functional class of the protein. This study suggests that the protein-folding problem has no unique solution and should be limited to defining the folding class of the solid-like fragments even though they may constitute only a small part of the protein. These results limit the capability of modeling protein structures with multiple conformational states. PMID- 19553206 TI - Old substrates for new enzymes of terpenoid biosynthesis. PMID- 19553208 TI - The Rayleigh-Taylor condition for the evolution of irrotational fluid interfaces. AB - For the free boundary dynamics of the two-phase Hele-Shaw and Muskat problems, and also for the irrotational incompressible Euler equation, we prove existence locally in time when the Rayleigh-Taylor condition is initially satisfied for a 2D interface. The result for water waves was first obtained by Wu in a slightly different scenario (vanishing at infinity), but our approach is different because it emphasizes the active scalar character of the system and does not require the presence of gravity. PMID- 19553207 TI - Changing meaning causes coupling changes within higher levels of the cortical hierarchy. AB - Processing of speech and nonspeech sounds occurs bilaterally within primary auditory cortex and surrounding regions of the superior temporal gyrus; however, the manner in which these regions interact during speech and nonspeech processing is not well understood. Here, we investigate the underlying neuronal architecture of the auditory system with magnetoencephalography and a mismatch paradigm. We used a spoken word as a repeating "standard" and periodically introduced 3 "oddball" stimuli that differed in the frequency spectrum of the word's vowel. The closest deviant was perceived as the same vowel as the standard, whereas the other 2 deviants were perceived as belonging to different vowel categories. The neuronal responses to these vowel stimuli were compared with responses elicited by perceptually matched tone stimuli under the same paradigm. For both speech and tones, deviant stimuli induced coupling changes within the same bilateral temporal lobe system. However, vowel oddball effects increased coupling within the left posterior superior temporal gyrus, whereas perceptually equivalent nonspeech oddball effects increased coupling within the right primary auditory cortex. Thus, we show a dissociation in neuronal interactions, occurring at both different hierarchal levels of the auditory system (superior temporal versus primary auditory cortex) and in different hemispheres (left versus right). This hierarchical specificity depends on whether auditory stimuli are embedded in a perceptual context (i.e., a word). Furthermore, our lateralization results suggest left hemisphere specificity for the processing of phonological stimuli, regardless of their elemental (i.e., spectrotemporal) characteristics. PMID- 19553209 TI - Whole-proteome phylogeny of large dsDNA virus families by an alignment-free method. AB - The vast sequence divergence among different virus groups has presented a great challenge to alignment-based sequence comparison among different virus families. Using an alignment-free comparison method, we construct the whole-proteome phylogeny for a population of viruses from 11 viral families comprising 142 large dsDNA eukaryote viruses. The method is based on the feature frequency profiles (FFP), where the length of the feature (l-mer) is selected to be optimal for phylogenomic inference. We observe that (i) the FFP phylogeny segregates the population into clades, the membership of each has remarkable agreement with current classification by the International Committee on the Taxonomy of Viruses, with one exception that the mimivirus joins the phycodnavirus family; (ii) the FFP tree detects potential evolutionary relationships among some viral families; (iii) the relative position of the 3 herpesvirus subfamilies in the FFP tree differs from gene alignment-based analysis; (iv) the FFP tree suggests the taxonomic positions of certain "unclassified" viruses; and (v) the FFP method identifies candidates for horizontal gene transfer between virus families. PMID- 19553210 TI - Growing up blind does not change the neural bases of Theory of Mind. AB - Humans reason about the mental states of others; this capacity is called Theory of Mind (ToM). In typically developing adults, ToM is supported by a consistent group of brain regions: the bilateral temporoparietal junction (TPJ), medial prefrontal cortex (MPFC), precuneus (PC), and anterior temporal sulci (aSTS). How experience and intrinsic biological factors interact to produce this adult functional profile is not known. In the current study we investigate the role of visual experience in the development of the ToM network by studying congenitally blind adults. In experiment 1, participants listened to stories and answered true/false questions about them. The stories were either about mental or physical representations of reality (e.g., photographs). In experiment 2, participants listened to stories about people's beliefs based on seeing or hearing; people's bodily sensations (e.g., hunger); and control stories without people. Participants judged whether each story had positive or negative valance. We find that ToM brain regions of sighted and congenitally blind adults are similarly localized and functionally specific. In congenitally blind adults, reasoning about mental states leads to activity in bilateral TPJ, MPFC, PC, and aSTS. These brain regions responded more to passages about beliefs than passages about nonbelief representations or passages about bodily sensations. Reasoning about mental states that are based on seeing is furthermore similar in congenitally blind and sighted individuals. Despite their different developmental experience, congenitally blind adults have a typical ToM network. We conclude that the development of neural mechanisms for ToM depends on innate factors and on experiences represented at an abstract level, amodally. PMID- 19553212 TI - Universals and cultural variation in turn-taking in conversation. AB - Informal verbal interaction is the core matrix for human social life. A mechanism for coordinating this basic mode of interaction is a system of turn-taking that regulates who is to speak and when. Yet relatively little is known about how this system varies across cultures. The anthropological literature reports significant cultural differences in the timing of turn-taking in ordinary conversation. We test these claims and show that in fact there are striking universals in the underlying pattern of response latency in conversation. Using a worldwide sample of 10 languages drawn from traditional indigenous communities to major world languages, we show that all of the languages tested provide clear evidence for a general avoidance of overlapping talk and a minimization of silence between conversational turns. In addition, all of the languages show the same factors explaining within-language variation in speed of response. We do, however, find differences across the languages in the average gap between turns, within a range of 250 ms from the cross-language mean. We believe that a natural sensitivity to these tempo differences leads to a subjective perception of dramatic or even fundamental differences as offered in ethnographic reports of conversational style. Our empirical evidence suggests robust human universals in this domain, where local variations are quantitative only, pointing to a single shared infrastructure for language use with likely ethological foundations. PMID- 19553211 TI - Reliable neuromodulation from circuits with variable underlying structure. AB - Recent work argues that similar network performance can result from highly variable sets of network parameters, raising the question of whether neuromodulation can be reliable across individuals with networks with different sets of synaptic strengths and intrinsic membrane conductances. To address this question, we used the dynamic clamp to construct 2-cell reciprocally inhibitory networks from gastric mill (GM) neurons of the crab stomatogastric ganglion. When the strength of the artificial inhibitory synapses (g(syn)) and the conductance of an artificial I(h) (g(h)) were varied with the dynamic clamp, a variety of network behaviors resulted, including regions of stable alternating bursting. Maps of network output as a function of g(syn) and g(h) were constructed in normal saline and again in the presence of serotonin or oxotremorine. Both serotonin and oxotremorine depolarize and excite isolated individual GM neurons, but by different cellular mechanisms. Serotonin and oxotremorine each increased the size of the parameter regions that supported alternating bursting, and, on average, increased burst frequency. Nonetheless, in both cases some parameter sets within the sample space deviated from the mean population response and decreased in frequency. These data provide insight into why pharmacological treatments that work in most individuals can generate anomalous actions in a few individuals, and they have implications for understanding the evolution of nervous systems. PMID- 19553213 TI - Dynamics of a vesicle in general flow. AB - An approach to quantitatively study vesicle dynamics as well as biologically related micro-objects in a fluid flow, which is based on the combination of a dynamical trap and a control parameter, the ratio of the vorticity to the strain rate, is suggested. The flow is continuously varied between rotational, shearing, and elongational in a microfluidic 4-roll mill device, the dynamical trap, that allows scanning of the entire phase diagram of motions, i.e., tank-treading (TT), tumbling (TU), and trembling (TR), using a single vesicle even at lambda = eta(in)/eta(out) = 1, where eta(in) and eta(out) are the viscosities of the inner and outer fluids. This cannot be achieved in pure shear flow, where the transition between TT and either TU or TR is attained only at lambda>1. As a result, it is found that the vesicle dynamical states in a general are presented by the phase diagram in a space of only 2 dimensionless control parameters. The findings are in semiquantitative accord with the recent theory made for a quasi spherical vesicle, although vesicles with large deviations from spherical shape were studied experimentally. The physics of TR is also uncovered. PMID- 19553214 TI - Single-particle cryoEM reconstructions: meeting the challenge. PMID- 19553215 TI - Profile of Raghavendra Gadagkar. PMID- 19553216 TI - Continuous dissolution of structure during the unfolding of a small protein. AB - The unfolding kinetics of many small proteins appears to be first order, when measured by ensemble-averaging probes such as fluorescence and circular dichroism. For one such protein, monellin, it is shown here that hidden behind this deceptive simplicity is a complexity that becomes evident with the use of experimental probes that are able to discriminate between different conformations in an ensemble of structures. In this study, the unfolding of monellin has been probed by measurement of the changes in the distributions of 4 different intramolecular distances, using a multisite, time-resolved fluorescence resonance energy transfer methodology. During the course of unfolding, the protein molecules are seen to undergo slow and continuous, diffusive swelling. The swelling process can be modeled as the slow diffusive swelling of a Rouse-like chain with some additional noncovalent, intramolecular interactions. Here, we show that specific structure is lost during the swelling process gradually, and not in an all-or-none manner, during unfolding. PMID- 19553217 TI - Linking molecular affinity and cellular specificity in cadherin-mediated adhesion. AB - Many cell-cell adhesive events are mediated by the dimerization of cadherin proteins presented on apposing cell surfaces. Cadherin-mediated processes play a central role in the sorting of cells into separate tissues in vivo, but in vitro assays aimed at mimicking this behavior have yielded inconclusive results. In some cases, cells that express different cadherins exhibit homotypic cell sorting, forming separate cell aggregates, whereas in other cases, intermixed aggregates are formed. A third pattern is observed for mixtures of cells expressing either N- or E-cadherin, which form distinct homotypic aggregates that adhere to one another through a heterotypic interface. The molecular basis of cadherin-mediated cell patterning phenomena is poorly understood, in part because the relationship between cellular adhesive specificity and intermolecular binding free energies has not been established. To clarify this issue, we have measured the dimerization affinities of N-cadherin and E-cadherin. These proteins are similar in sequence and structure, yet are able to mediate homotypic cell patterning behavior in a variety of tissues. N-cadherin is found to form homodimers with higher affinity than does E-cadherin and, unexpectedly, the N/E cadherin heterophilic binding affinity is intermediate in strength between the 2 homophilic affinities. We can account for observed cell aggregation behaviors by using a theoretical framework that establishes a connection between molecular affinities and cell-cell adhesive specificity. Our results illustrate how graded differences between different homophilic and heterophilic cadherin dimerizaton affinities can result in homotypic cell patterning and, more generally, show how proteins that are closely related can, nevertheless, be responsible for highly specific cellular adhesive behavior. PMID- 19553218 TI - A biomarker based on the stable isotopes of nickel. AB - The new stable isotope systems of transition metals are increasingly used to understand and quantify the impact of primitive microbial metabolisms on the modern and ancient Earth. To date, little effort has been expended on nickel (Ni) isotopes but there are good reasons to believe that this system may be more straightforward, and useful in this respect, than some others. Here, we present Ni stable isotope data for abiotic terrestrial samples and pure cultures of methanogens. The dataset for rocks reveals little isotopic variability and provides a lithologic baseline for terrestrial Ni isotope studies. In contrast, methanogens assimilate the light isotopes, yielding residual media with a complementary heavy isotopic enrichment. Methanogenesis may have evolved during or before the Archean, when methane could have been key to Earth's early systems. Our data suggest significant potential in Ni stable isotopes for identifying and quantifying methanogenesis on the early planet. Additionally, Ni stable isotope fractionation may well prove to be the fundamental unambiguous trace metal biomarker for methanogens. PMID- 19553219 TI - GFP: Lighting up life. PMID- 19553220 TI - Exercise is beneficial for patients with Alzheimer's disease: a call for action. PMID- 19553221 TI - Five-year prospective comparison study of topical glyceryl trinitrate treatment of chronic lateral epicondylosis at the elbow. AB - BACKGROUND: Topical glyceryl trinitrate (GTN) treatment has previously demonstrated short-term efficacy in the treatment of lateral epicondylosis. No long-term follow-up has been performed. HYPOTHESIS: Benefits from topical GTN persist 5 years after the cessation of therapy. STUDY DESIGN: A prospective comparative study. METHODS: A follow-up study of 58 patients treated with 6 months of topical GTN or placebo, combined with a tendon rehabilitation programme, was performed 5 years after discontinuation of therapy. Assessment included patient-rated pain scores, clinically assessed lateral epicondylar and proximal common extensor tendon tenderness, hand-held dynamometer measurement of resisted third finger metacarpophalangeal extension with a fully extended elbow (Maudsley's test) and wrist extensor tendon mean peak force using a modified chair pick-up test (the Orthopaedic Research Institute-tennis elbow testing system). RESULTS: Patients in both the GTN group and those in the placebo group had significant improvements in symptoms, clinical signs and provocative functional tests compared with baseline week 0 measures. GTN did not offer any additional clinical benefit over a standard tendon rehabilitation programme at 5 years. CONCLUSION: While GTN appears to offer short-term benefits up to 6 months in the treatment of lateral epicondylosis, at 5 years there does not appear to be significant clinical benefits when compared with patients undertaking a standard tendon rehabilitation programme alone. This is in contrast to findings of continued benefits at long-term follow-up described in the literature for patients with Achilles tendinopathy treated with GTN. PMID- 19553222 TI - Tactile acuity and lumbopelvic motor control in patients with back pain and healthy controls. AB - BACKGROUND: Voluntary lumbopelvic control is compromised in patients with back pain. Loss of proprioceptive acuity is one contributor to decreased control. Several reasons for decreased proprioceptive acuity have been proposed, but the integrity of cortical body maps has been overlooked. We investigated whether tactile acuity, a clear clinical signature of primary sensory cortex organisation, relates to lumbopelvic control in people with back pain. METHODS: Forty-five patients with back pain and 45 age- and sex-matched healthy controls participated in this cross-sectional study. Tactile acuity at the back was assessed using two-point discrimination (TPD) threshold in vertical and horizontal directions. Voluntary motor control was assessed using an established battery of clinical tests. RESULTS: Patients performed worse on the voluntary lumbopelvic tasks than healthy controls did (p<0.001). TPD threshold was larger in patients (mean (SD)=61 (13) mm) than in healthy controls (44 (10) mm). Moreover, larger TPD threshold was positively related to worse performance on the voluntary lumbopelvic tasks (Pearson's r=0.49; p<0.001). DISCUSSION: Tactile acuity, a clear clinical signature of primary sensory cortex organisation, relates to voluntary lumbopelvic control. This relationship raises the possibility that the former contributes to the latter, in which case training tactile acuity may aid recovery and assist in achieving normal motor performance after back injury. PMID- 19553223 TI - Rugby league injuries in New Zealand: a review of 8 years of Accident Compensation Corporation injury entitlement claims and costs. AB - AIM: This paper provides an overview of the epidemiology of rugby league injuries and associated costs in New Zealand requiring medical treatment. METHOD: New Zealand national Accident Compensation Corporation injury data for the period 1999 to 2007 were searched for rugby league injury cases. Data were analysed by demographics, body region, nature/severity of injury, and medical procedure and costs. RESULTS: A total of 5941 injury entitlement claims were recorded over the study period with a significant decrease observed in the injury rate between the 1999-2000 and 2002-2003 reporting years. The total cost of the injuries for the study period was $42,822,048 (equivalent to pound15,916,072). The mean (SD) number of injury entitlement claims per year was 743 (271) and yearly cost was $5,352,760 (pound1,989,880) ($2,485,535 (pound923,994)). The knee was the most commonly reported injury site (225 per 1000 entitlement claims; $8,750,147 (pound3,252,020)) and soft tissue injuries were the most common injury types (474 per 1000 entitlement claims; $17,324,214 (pound6,438,599)). Accounting for only 1.8% of total injury entitlement claims, concussion/brain injuries accounted for 6.3% of injury entitlement costs and had the highest mean cost per claim ($25 347 (pound9420)). The upper and lower arm recorded the highest mean injury site claim cost of $43,096 (pound16,016) per claim. The 25-29 age group recorded 27.7% of total injury entitlement claims and 29.6% of total injury entitlement costs, which was slightly more than the 20-24 age group (27.3% claims; 24.7% costs). Nearly 15% of total moderate to serious injury entitlement claims and 20% of total costs were recorded from participants 35 years or older. DISCUSSION: This study identified that the knee was the most common injury site and soft tissue injuries were the most common injury type requiring medical treatment, which is consistent with other international studies on rugby league epidemiology. This study also highlights that the rate of injury and the average age of injured rugby league players increased over time. The high cost of concussion/brain injuries is a cause for concern as it reflects the severity of the injuries. CONCLUSION: Injury prevention programmes for rugby league should focus on reducing the risk of concussion/brain injury and knee and soft tissue injury, and should target participants in the 20-30 years old age range. More longitudinal epidemiological studies with specific details on injury mechanisms and participation data are warranted to further identify the injury circumstances surrounding participation in rugby league activities. PMID- 19553224 TI - Trp64Arg polymorphism in ADRB3 gene is associated with elite endurance performance. AB - In this study, allele and genotype frequencies of the ADRB1 Arg389Gly (rs1801253), ADRB2 Gly16Arg (rs1042713) and Gln27Glu (rs1042714), and ADRB3 Trp64Arg (rs4994) variations were compared in the following three groups of Spanish (Caucasian) men: (1) world-class endurance athletes (E; runners and cyclists, n=100), (2) elite power athletes (P; sprinters, jumpers and throwers, n=53) and (3) non-athletic controls (C; n=100). No significant differences were observed in genotype and allele distributions among the study groups except for the ADRB3 Trp64Arg polymorphism in E versus C (27% vs 8% of carriers of the Arg allele in E and C, p<0.001; frequency of the minor Arg (C) allele of 14% vs 4% in E and C, p=0.001). Heterozygosity for the ADRB3 Trp64Arg polymorphism seems to be associated with elite endurance performance, while other variants of the beta adrenergic receptors' genes do not seem to significantly influence top-level sports performance, at least in athletes of Spanish origin. PMID- 19553225 TI - Injury incidence and injury patterns in professional football: the UEFA injury study. AB - OBJECTIVE: To study the injury characteristics in professional football and to follow the variation of injury incidence during a match, during a season and over consecutive seasons. DESIGN: Prospective cohort study where teams were followed for seven consecutive seasons. Team medical staff recorded individual player exposure and time-loss injuries from 2001 to 2008. SETTING: European professional men's football. PARTICIPANTS: The first team squads of 23 teams selected by the Union of European Football Associations as belonging to the 50 best European teams. MAIN OUTCOME MEASUREMENT: Injury incidence. RESULTS: 4483 injuries occurred during 566 000 h of exposure, giving an injury incidence of 8.0 injuries/1000 h. The injury incidence during matches was higher than in training (27.5 vs 4.1, p<0.0001). A player sustained on average 2.0 injuries per season, and a team with typically 25 players can thus expect about 50 injuries each season. The single most common injury subtype was thigh strain, representing 17% of all injuries. Re-injuries constituted 12% of all injuries, and they caused longer absences than non re-injuries (24 vs 18 days, p<0.0001). The incidence of match injuries showed an increasing injury tendency over time in both the first and second halves (p<0.0001). Traumatic injuries and hamstring strains were more frequent during the competitive season, while overuse injuries were common during the preseason. Training and match injury incidences were stable over the period with no significant differences between seasons. CONCLUSIONS: The training and match injury incidences were stable over seven seasons. The risk of injury increased with time in each half of matches. PMID- 19553226 TI - The role of emotions on pacing strategies and performance in middle and long duration sport events. AB - Thepacing strategy may be defined as the process in which the total energy expenditure during exercise is regulated on a moment-to-moment basis in order to ensure that the exercise bout can be completed in a minimum time and without a catastrophic biological failure. Experienced athletes develop a stable template of the power outputs they are able to sustain for different durations of exercise, but it is not known how they originally develop this template or how that template changes with training and experience. While it is understood that the athlete's physiological state makes an important contribution to this process, there has been much less interest in the contribution that the athlete's emotional status makes. The aim of this review is to evaluate the literature of physiological, neurophysiological and perceptual responses during exercise in order to propose a complex model interpretation of this process which may be a critical factor determining success in middle- and long-duration sporting competitions. We describe unconscious/physiological and conscious/emotional mechanisms of control, the focus of which are to ensure that exercise terminates before catastrophic failure occurs in any bodily system. We suggest that training sessions teach the athlete to select optimal pacing strategies by associating a level of emotion with the ability to maintain that pace for exercise of different durations. That pacing strategy is then adopted in future events. Finally, we propose novel perspectives to maximise performance and to avoid overtraining by paying attention also to the emotional state in training process. PMID- 19553227 TI - Genetic risk factors for soft-tissue injuries 101: a practical summary to help clinicians understand the role of genetics and 'personalised medicine'. PMID- 19553229 TI - Cancer incidence and mortality in a historical cohort of Australian pest control workers. AB - OBJECTIVES: To determine the rates of mortality and incident cancer in a historical cohort of pest control operators. METHODS: A retrospective cohort was assembled from former state government occupational health surveillance programmes. This cohort was linked to the Australian national registries of cancer and mortality and the results were compared with the general Australian population rates. RESULTS: 125 deaths and 89 incident cancers were found during the periods of observation (mortality 1983-2004 and cancer 1983-2002). Overall cancer incidence and mortality rates were not found to be significantly different from the general population. Among the specific causes of death, suicide (standardised mortality ratio; SMR 1.78; 95% CI 1.12 to 2.83) and unintentional falls (SMR 4.57; 95% CI 1.72 to 12.19) were significantly in excess, although the latter was based on only 4 deaths. Melanoma was the only specific incident cancer found significantly in excess (standardised incidence ratio 1.56; 95% CI 1.03 to 2.37). CONCLUSIONS: Pest control workers have overall mortality and cancer rates similar to the general population. Excess rates of incident melanoma and intentional self-poisoning mortality are of concern and warrant further investigation. Follow-up of this cohort as its members age, will provide more insight into these possible associations. PMID- 19553228 TI - Traffic particles and occurrence of acute myocardial infarction: a case-control analysis. AB - OBJECTIVES: We modelled exposure to traffic particles using a latent variable approach and investigated whether long-term exposure to traffic particles is associated with an increase in the occurrence of acute myocardial infarction (AMI) using data from a population-based coronary disease registry. METHODS: Cases of individually validated AMI were identified between 1995 and 2003 as part of the Worcester Heart Attack Study. Population controls were selected from Massachusetts, USA, resident lists. NO(2) and PM(2.5) filter absorbance were measured at 36 locations throughout the study area. The air pollution data were used to estimate exposure to traffic particles using a semiparametric latent variable regression model. Conditional logistic models were used to estimate the association between exposure to traffic particles and occurrence of AMI. RESULTS: Modelled exposure to traffic particles was highest near the city of Worcester. Cases of AMI were more exposed to traffic and traffic particles compared to controls. An interquartile range increase in modelled traffic particles was associated with a 10% (95% CI 4% to 16%) increase in the odds of AMI. Accounting for spatial dependence at the census tract, but not block group, scale substantially attenuated this association. CONCLUSIONS: These results provide some support for an association between long-term exposure to traffic particles and risk of AMI. The results were sensitive to the scale selected for the analysis of spatial dependence, an issue that requires further investigation. The latent variable model captured variation in exposure, although on a relatively large spatial scale. PMID- 19553230 TI - Ischaemic heart disease mortality study among workers with occupational exposure to ammonium perfluorooctanoate. AB - OBJECTIVES: Ammonium perfluorooctanoate (APFO) is a biopersistent surfactant used in the manufacture of several types of fluoropolymers. Based on previous findings of increased serum lipid levels associated with exposure to APFO, we evaluated ischaemic heart disease (IHD) mortality in a cohort of occupationally exposed workers. METHODS: Relative risks (RR) were estimated from exposure-response analyses of cumulative exposure measures using proportional hazards regression models. RESULTS: 239 IHD deaths have occurred in the cohort of 4747 workers with work histories from 1948 through 2002. RR estimates indicate no statistically significant increased mortality risk for IHD associated with estimated cumulative exposure. We observed a positive trend only at an exposure lag of 10 years. This finding was not reproduced in other 5-year exposure lags and was attenuated when different cutpoints for exposure categorisation were used. CONCLUSION: This exposure-response study shows no convincing evidence of increased IHD mortality risk for APFO-exposed workers at this plant. Further studies evaluating the incidence of IHD are being conducted. PMID- 19553231 TI - The diagnostic value of provocative clinical tests in ulnar neuropathy at the elbow is marginal. AB - BACKGROUND: Provocative clinical tests are often performed in the diagnosis of ulnar neuropathy at the elbow (UNE) although the evidence for the usefulness of these tests is limited. The aim of this study was to determine the diagnostic value of provocative clinical tests in the diagnosis of UNE in a relevant spectrum of patients and controls. METHODS: A prospective cohort study was performed in consecutive patients clinically suspected of having UNE. All patients underwent a neurological examination and four commonly used provocative clinical tests (Tinel's test, flexion compression test, palpating for local ulnar nerve tenderness and nerve thickening). Subsequently, in all patients a reference standard test comprising electrophysiological studies and neurosonography was independently assessed. RESULTS: 192 eligible patients completed the study protocol. UNE was diagnosed in 137 and an alternative diagnosis was made in 55 patients. The sensitivity, specificity, and positive and negative predictive values were as follows: Tinel's test 62%, 53%, 77% and 30%; flexion compression test 61%, 40%, 72% and 29%; palpating for nerve thickening 28%, 87%, 84% and 33%; and palpating for nerve tenderness 32%, 80%, 80% and 32%. Logistic regression and receiver operating characteristic curves showed that the added value of one or more provocative tests over routine clinical examination is minimal. CONCLUSION: The diagnostic value of provocative clinical tests in UNE is poor. PMID- 19553232 TI - Commentary: How does 'insurance' improve equity in health? PMID- 19553233 TI - Prevalence of COPD in Spain: impact of undiagnosed COPD on quality of life and daily life activities. AB - AIMS: This study aimed to determine the prevalence of chronic obstructive pulmonary disease (COPD) in Spain and identify the level of undiagnosed disease and its impact on health-related quality of life (HRQL) and activities of daily living (ADL). METHODS: A population-based sample of 4274 adults aged 40-80 years was surveyed. They were invited to answer a questionnaire and undergo prebrochodilator and postbronchodilator spirometry. COPD was defined as a postbronchodilator FEV(1)/FVC (forced expiratory volume in 1 s/forced vital capacity) ratio of <0.70. RESULTS: For 3802 participants with good-quality postbronchodilator spirometry, the overall prevalence of COPD was 10.2% (95% CI 9.2% to 11.1%) and was higher in men (15.1%) than in women (5.6%). The prevalence of COPD stage II or higher was 4.4% (95%CI; 3.8%-5.1%). The prevalence of COPD increased with age and with cigarette smoking and was higher in those with a low educational level. A previous diagnosis of COPD was reported by only 27% of those with COPD. Diagnosed patients had more severe disease, higher cumulative tobacco consumption and more severely impaired HRQL compared with undiagnosed subjects. However, even patients with undiagnosed COPD stage I+ already showed impairment in HRQL and in some aspects of ADL compared with participants without COPD. CONCLUSIONS: The prevalence of COPD in individuals between 40 and 80 years of age in Spain is 10.2% and increases with age, tobacco consumption and lower educational levels. The rate of diagnosised COPD is very high and undiagnosed individuals with COPD already have a significant impairment in HRQL and ADL. PMID- 19553234 TI - Association between mycobacterial genotypes and disease progression in Mycobacterium avium pulmonary infection. AB - BACKGROUND: Non-tuberculous mycobacterial lung disease, most commonly caused by Mycobacterium avium infection, tends to show variable disease progression, and significant disease predictors have not been adequately established. METHODS: Variable numbers of tandem repeats (VNTR) were evaluated in 16 mycobacterial interspersed repetitive unit (MIRU) loci from M avium isolates cultured from respiratory specimens obtained from 2005 to 2007. Specifically, the association between VNTR profiles and disease progression was assessed. RESULTS: Among the 37 subjects who provided positive respiratory cultures for M avium during the 2005-6 period, 15 subjects were treated within 10 months following a microbiological diagnosis of progressive M avium lung disease. Nine subjects underwent long-term follow-up (>24 months) without treatment for stable M avium lung disease. Based on a neighbour-joining cluster analysis used to classify M avium-positive subjects according to the VNTR profile, subjects with progressive versus stable lung disease were found to be grouped together in distinct clusters. Further analysis using logistic regression modelling showed that disease progression was significantly associated with the genetic distance of the M avium isolate from an appropriately selected reference (age-adjusted odds ratio 1.95; 95% confidence interval 1.16 to 3.30; p = 0.01 for the most significant model). A best-fit model could be used to predict the progression of M avium lung disease when subjects from the 2005-6 period were combined with those from 2007 (p = 0.003). CONCLUSION: Progressive lung disease due to M avium infection is associated with specific VNTR genotypes of M avium. PMID- 19553235 TI - The externally placed 'temporary-permanent' generator. PMID- 19553238 TI - Exposure of Xenopus laevis tadpoles to finasteride, an inhibitor of 5-alpha reductase activity, impairs spermatogenesis and alters hypophyseal feedback mechanisms. AB - Sexual steroids have major regulatory functions in gonadal development, maturation of gametes and sexual differentiation in vertebrates. Previous studies in amphibians provided evidence that dihydrotestosterone and activity of 5-alpha reductases might play a significant role in androgen-mediated reproductive biology. To test the involvement of 5-alpha reductases in maturation of gametes in amphibians, Xenopus laevis was exposed to finasteride (FIN), a known inhibitor of 5-alpha reductase enzyme activity. In a long-term exposure from stage 46 to 66, severe disruption of spermatogenesis was observed in histological analysis of testes as detected by occurrence of empty spermatocysts, while ovaries remained unaffected. Real-time PCR analyses of male and female brain revealed an increase of LHbeta mRNA and a decrease of FSHbeta mRNA in males, suggesting a signalling on testes that could result in increased steroidogenesis and reduced Sertoli cell proliferation. Accordingly, the mRNA expression of P450 side chain cleavage enzyme and 5-alpha reductase type 2 was increased in testes, while no effects could be observed on steroidogenic genes in ovaries. A short-term exposure to testosterone, FIN and testosterone+FIN showed that transient effects of FIN targeted males selectively and, in particular, interfered with the hypothalamus pituitary-gonad axis. Furthermore, a negative feedback of testosterone on LHbeta was observed on males and females. This study provides evidence that exposure of X. laevis to FIN, an inhibitor of 5-alpha reductases, impaired spermatogenesis and involved sex-specific hypophyseal feedback mechanisms. PMID- 19553239 TI - Selection of sperm based on combined density gradient and Zeta method may improve ICSI outcome. AB - BACKGROUND: Reducing the percentage of sperm anomalies in insemination samples remains a goal to be achieved in the intra-cytoplasmic sperm insemination (ICSI) procedure. The aim of this study was to evaluate the efficiency of density gradient centrifugation (DGC) and Zeta methods to recover sperm with intact chromatin, and to evaluate whether the combined DGC/Zeta procedure improved ICSI outcome. METHODS: In Experiment 1, DGC and Zeta methods were carried out on 60 unprocessed semen samples. The samples were then assessed by chromomycin A3 staining, acridine orange test, terminal deoxynucleotidyl transferase dUTP nick end labeling assay and the sperm chromatin dispersion test for protamine deficiency and DNA fragmentation. In Experiment 2, sibling oocytes from 30 ICSI candidates were divided into two groups; one group was inseminated with sperm processed by DGC and the second with sperm processed by DGC/Zeta. The outcomes of 30 ICSI cycles were compared between the two groups and also with 34 ICSI candidates whose oocytes were inseminated by DGC-processed sperm. RESULTS: Both procedures were efficient for the recovery of sperm with normal protamine content and low DNA fragmentation. However, the Zeta method yielded a greater number of sperm with less DNA fragmentation. Fertilization and pregnancy rates were improved following the combined DGC/Zeta procedure. Compared with DGC alone, the pregnancy rate appeared improved but this was not statistically significant (P = 0.091). CONCLUSIONS: Combining DGC and Zeta procedures improves the quality of semen samples which may increase fertilization rates and possibly pregnancy rates. PMID- 19553237 TI - Inverse effects on gating and modulation caused by a mutation in the M2-M3 Linker of the GABA(A) receptor gamma subunit. AB - M2-M3 linkers are receptor subunit domains known to be critical for the normal function of cysteine-loop ligand-gated ion channels. Previous studies of alpha and beta subunits of type "A" GABA receptors suggest that these linkers couple extracellular elements involved in GABA binding to the transmembrane segments that control the opening of the ion channel. To study the importance of the gamma subunit M2-M3 linker, we examined the macroscopic and single-channel effects of an engineered gamma2(L287A) mutation on GABA activation and propofol modulation. In the macroscopic analysis, we found that the gamma2(L287A) mutation decreased GABA potency but increased the ability of propofol to enhance both GABA potency and efficacy compared with wild-type receptors. Indeed, although propofol had significant effects on GABA potency in wild-type receptors, we found that propofol produced no corresponding increase in GABA efficacy. At the single channel level, mutant receptors showed a loss in the longest of three open-time components compared with wild-type receptors under GABA activation. Furthermore, propofol reduced the duration of one closed-time component, increased the duration of two open-time components, and generated a third open component with a longer lifetime in mutant compared with wild-type receptors. Taken together, we conclude that although the gamma subunit is not required for the binding of GABA or propofol, the M2-M3 linker of this subunit plays a critical role in channel gating by GABA and allosteric modulation by propofol. Our results also suggest that in wild-type receptors, propofol exerts its enhancing effects by mechanisms extrinsic to channel gating. PMID- 19553236 TI - Serotonin shapes risky decision making in monkeys. AB - Some people love taking risks, while others avoid gambles at all costs. The neural mechanisms underlying individual variation in preference for risky or certain outcomes, however, remain poorly understood. Although behavioral pathologies associated with compulsive gambling, addiction and other psychiatric disorders implicate deficient serotonin signaling in pathological decision making, there is little experimental evidence demonstrating a link between serotonin and risky decision making, in part due to the lack of a good animal model. We used dietary rapid tryptophan depletion (RTD) to acutely lower brain serotonin in three macaques performing a simple gambling task for fluid rewards. To confirm the efficacy of RTD experiments, we measured total plasma tryptophan using high-performance liquid chromatography (HPLC) with electrochemical detection. Reducing brain serotonin synthesis decreased preference for the safe option in a gambling task. Moreover, lowering brain serotonin function significantly decreased the premium required for monkeys to switch their preference to the risky option, suggesting that diminished serotonin signaling enhances the relative subjective value of the risky option. These results implicate serotonin in risk-sensitive decision making and, further, suggest pharmacological therapies for treating pathological risk preferences in disorders such as problem gambling and addiction. PMID- 19553240 TI - The number of oogonia and somatic cells in the human female embryo and fetus in relation to whether or not exposed to maternal cigarette smoking. AB - BACKGROUND: Prenatal exposure to maternal cigarette smoking or compounds of cigarette smoke is associated with serious reproductive hazards such as apoptotic death of oogonia in murine offspring and decreased fecundability in human offspring. The present study addresses potential effects of in utero exposure to cigarette smoking. METHODS: Twenty-nine human first-trimester ovaries from legal abortions [aged 38-64 days post-conception (p.c.)] were collected. Mothers filled out a questionnaire about their smoking habits and delivered a urine sample for cotinine analysis. The ovarian cell numbers were estimated using stereological methods. RESULTS: A non-linear correlation between the numbers of oogonia and somatic cells in relation to age of the embryo/fetus was shown in 28 ovaries, including the first estimates performed in ovaries younger than 47 days p.c. Prenatal exposure to smoke showed a significant decrease in the number of somatic cells (P < or = 0.01). The number of oogonia was not significantly associated with prenatal exposure to maternal smoking (P < or = 0.09). The ratio between the two cell types decreased considerably from 1:45 to 1:23 from 38 to 46 days p.c. and was not affected by smoking. CONCLUSIONS: Oogonia proliferate and/or invade the developing ovary at a much faster relative rate than somatic cells. In utero exposure to maternal smoking significantly reduces the number of somatic cells from Days 38 to 64 p.c. Since oocytes cannot survive without being enclosed by somatic cells in a follicle, reduction in the somatic cells number may have long range consequences on the number of oocytes available in adult life and on the future fertility of female offspring exposed to smoking in utero. PMID- 19553241 TI - Recruitment bias in studies of semen and other factors affecting pregnancy rates in fertile men. AB - BACKGROUND: Recruitment bias is possible in population studies of semen quality because few men volunteer. We examine differences between Australian couples with natural conceptions who agreed or declined to participate in such a study. METHODS: Women pregnant between 16 and 32 weeks gestation participating in a retrospective time to pregnancy (TTP) study were each requested to recruit their eligible (on the basis of age, place of his birth and of his mother's birth) male partner to complete additional questionnaires, have a physical examination and provide blood and two semen samples. RESULTS: From 2061 women who completed the TTP questionnaire (response rate, 98%) there were 928 eligible male partners of whom 225 (24%) were responders. There were significant socio-demographic and self reported exposure differences between responders and non-responders in particular, female professional occupation, knowledge of the fertile phase, pelvic inflammatory disease, non-smoker at time of conception and wine consumption per week were more frequent in the responders. There was no evidence of a bias for the subfertile being more likely to volunteer for the study. Mean TTP for planned pregnancies for responders and non-responders were 3.3 and 3.8 cycles (P = 0.319), respectively, and the cycle specific pregnancy rates were not significantly different after covariate adjustment by Cox regression. CONCLUSIONS: The present study confirms that participation rates are low in studies of semen quality. Although the expected higher participation of subfertile couples was not confirmed, there remains considerable potential for bias and other problems that could invalidate this type of study. PMID- 19553242 TI - Identification of a novel human cellular HDL biosynthesis defect. AB - AIMS: Severe high-density lipoprotein cholesterol (HDL-C) deficiency is attributed to mutations in several genes and may contribute to the genetic basis of coronary artery disease. To identify the cellular basis of a novel HDL deficiency phenotype, we screened 54 subjects of French Canadian ancestry with severe HDL deficiency. METHODS AND RESULTS: We excluded individuals with mutations in genes currently associated with low HDL (ABCA1, LCAT, APOA-I, and SMPD1). We identified two patients in which cellular phospholipid efflux in the HDL biosynthesis process is impaired, whereas cholesterol efflux is normal. Two dimensional gel electrophoresis analysis further showed that the two patients with impaired phospholipid efflux were defective primarily in the larger alpha HDL subpopulations. In fibroblasts from affected subjects, oxysterol stimulation resulted in increased ABCA1 protein expression and normalized their defective phospholipid efflux defect. CONCLUSION: Our results indicate for the first time in humans that phospholipid and cholesterol efflux are two separate and distinct processes in cellular HDL biosynthesis. They further show for the first time that normal cellular phospholipid efflux is necessary for the formation of larger alpha-HDL particles. The defect in phospholipid efflux is due to defective ABCA1 protein regulation and can be corrected by treatment with physiological oxysterols, a current therapeutic target of interest, that may, with further studies, be used to raise HDL levels in patients with severe HDL deficiencies. PMID- 19553243 TI - Coronary and vertebral subclavian steal demonstrated by subclavian angiography. PMID- 19553246 TI - Capitalizing on nature: how to implement an ecosystem approach. AB - The Natural Capital Initiative (www.naturalcapitalinitiative.org.uk) held its first conference 'Valuing our life support systems' at Savoy Place, London, from 29 April to 1 May 2009. The aim of the conference was to discuss different perspectives on, and solutions to, the conservation and sustainable use of ecosystem services. It particularly focused on the link between the environment and the economy, and how to implement an ecosystem approach to environmental management. This event brought together scientists across the natural and social sciences, alongside representatives from government, non-governmental organizations, business and industry. PMID- 19553247 TI - Differences in the 2nd to 4th digit length ratio in humans reflect shifts along the common allometric line. AB - Ratios often lead to biased conclusions concerning the actual relationships between examined traits and comparisons of the relative size of traits among groups. Therefore, the use of ratios has been abandoned in most comparative studies. However, ratios such as body mass index and waist-to-hip ratio are widely used in evolutionary biology and medicine. One such, the ratio of the 2nd to the 4th finger (2D : 4D), has been the subject of much recent interest in both humans and animals. Most studies agree that 2D : 4D is sexually dimorphic. In men, the 2nd digit tends to be shorter than the 4th, while in women the 2nd digit tends to be of the same size or slightly longer than the 4th. Nevertheless, here we demonstrate that the sexes do not greatly differ in the scaling between the 2nd and 4th digit. Sexual differences in 2D : 4D are mainly caused by the shift along the common allometric line with non-zero intercept, which means 2D : 4D necessarily decreases with increasing finger length, and the fact that men have longer fingers than women. We conclude that previously published results on the 2D : 4D ratio are biased by its covariation with finger length. We strongly recommend regression-based approaches for comparisons of hand shape among different groups. PMID- 19553248 TI - How does breeding system variation modulate sexual antagonism? AB - The study of sexually antagonistic (SA) traits remains largely limited to dioecious (separate sex), mobile animals. However, the occurrence of sexual conflict is restricted neither by breeding system (the mode of sexual reproduction, e.g. dioecy or hermaphroditism) nor by sessility. Here, we synthesize how variation in breeding system can affect the evolution and expression of intra- and inter-locus sexual conflicts in plants and animals. We predict that, in hermaphrodites, SA traits will (i) display lower levels of polymorphism; (ii) respond more quickly to selection; and (iii) involve unique forms of interlocus conflict over sex allocation, mating roles and selfing rates. Explicit modelling and empirical tests in a broader range of breeding systems are necessary to obtain a general understanding of the evolution of SA traits. PMID- 19553249 TI - Pitch then power: limitations to acceleration in quadrupeds. AB - Rapid acceleration and deceleration are vital for survival in many predator and prey animals and are important attributes of animal and human athletes. Adaptations for acceleration and deceleration are therefore likely to experience strong selective pressures--both natural and artificial. Here, we explore the mechanical and physiological constraints to acceleration. We examined two elite athletes bred and trained for acceleration performance (polo ponies and racing greyhounds), when performing maximal acceleration (and deceleration for ponies) in a competitive setting. We show that maximum acceleration and deceleration ability may be accounted for by two simple limits, one mechanical and one physiological. At low speed, acceleration and deceleration may be limited by the geometric constraints of avoiding net nose-up or tail-up pitching, respectively. At higher speeds, muscle power appears to limit acceleration. PMID- 19553250 TI - The non-invasive biopsy--will urinary proteomics make the renal tissue biopsy redundant? AB - Proteomics is a rapidly advancing technique which gives functional insight into gene expression in living organisms. Urine is an ideal medium for study as it is readily available, easily obtained and less complex than other bodily fluids. Considerable progress has been made over the last 5 years in the study of urinary proteomics as a diagnostic tool for renal disease. Advantages over the traditional renal biopsy include accessibility, safety, the possibility of serial sampling and the potential for non-invasive prognostic and diagnostic monitoring of disease and an individual's response to treatment. Urinary proteomics is now moving from a discovery phase in small studies to a validation phase in much larger numbers of patients with renal disease. Whilst there are still some limitations in methodology, which are assessed in this review, the possibility of urinary proteomics replacing the invasive tissue biopsy for diagnosis of renal disease is becoming an increasingly realistic option. PMID- 19553251 TI - Size-dependent alternative male mating tactics in the yellow dung fly, Scathophaga stercoraria. AB - Whenever males can monopolize females and/or resources used by females, the opportunity for sexual selection will be great. The greater the variation among males in reproductive success, the greater the intensity of selection on less competitive males to gain matings through alternative tactics. In the yellow dung fly, Scathophaga stercoraria, males aggressively compete for access to receptive, gravid females on fresh dung. Larger males are better able to acquire mates and to complete copulation successfully and guard the female throughout oviposition. Here we demonstrate that when an alternative resource is present where females aggregate (i.e. apple pomace, where both sexes come to feed), smaller males will redirect their searching for females from dung to the new substrate. In addition, we identify a class of particularly small males on the alternative substrate that appears never to be present searching for females on or around dung. Smaller males were found to have a mating 'advantage' on pomace, in striking contrast to the pattern observed on dung, providing further support for the existence of an alternative male reproductive tactic in this species. PMID- 19553252 TI - Brain organization mirrors caste differences, colony founding and nest architecture in paper wasps (Hymenoptera: Vespidae). AB - The cognitive challenges that social animals face depend on species differences in social organization and may affect mosaic brain evolution. We asked whether the relative size of functionally distinct brain regions corresponds to species differences in social behaviour among paper wasps (Hymenoptera: Vespidae). We measured the volumes of targeted brain regions in eight species of paper wasps. We found species variation in functionally distinct brain regions, which was especially strong in queens. Queens from species with open-comb nests had larger central processing regions dedicated to vision (mushroom body (MB) calyx collars) than those with enclosed nests. Queens from advanced eusocial species (swarm founders), who rely on pheromones in several contexts, had larger antennal lobes than primitively eusocial independent founders. Queens from species with morphologically distinct castes had augmented central processing regions dedicated to antennal input (MB lips) relative to caste monomorphic species. Intraspecific caste differences also varied with mode of colony founding. Independent-founding queens had larger MB collars than their workers. Conversely, workers in swarm-founding species with decentralized colony regulation had larger MB calyx collars and optic lobes than their queens. Our results suggest that brain organization is affected by evolutionary transitions in social interactions and is related to the environmental stimuli group members face. PMID- 19553253 TI - Sexual selection explains sex-specific growth plasticity and positive allometry for sexual size dimorphism in a reef fish. AB - In 1950, Rensch noted that in clades where males are the larger sex, sexual size dimorphism (SSD) tends to be more pronounced in larger species. This fundamental allometric relationship is now known as 'Rensch's rule'. While most researchers attribute Rensch's rule to sexual selection for male size, experimental evidence is lacking. Here, we suggest that ultimate hypotheses for Rensch's rule should also apply to groups of individuals and that individual trait plasticity can be used to test those hypotheses experimentally. Specifically, we show that in the sex-changing fish Parapercis cylindrica, larger males have larger harems with larger females, and that SSD increases with harem size. Thus, sexual selection for male body size is the ultimate cause of sexual size allometry. In addition, we experimentally illustrate a positive relationship between polygyny potential and individual growth rate during sex change from female to male. Thus, sexual selection is the ultimate cause of variation in growth rate, and variation in growth rate is the proximate cause of sexual size allometry. Taken together, our results provide compelling evidence in support of the sexual selection hypothesis for Rensch's rule and highlight the potential importance of individual growth modification in the shaping of morphological patterns in Nature. PMID- 19553254 TI - Forecasting the limits of resilience: integrating empirical research with theory. AB - Despite the increasing evidence of drastic and profound changes in many ecosystems, often referred to as regime shifts, we have little ability to understand the processes that provide insurance against such change (resilience). Modelling studies have suggested that increased variance may foreshadow a regime shift, but this requires long-term data and knowledge of the functional links between key processes. Field-based research and ground-truthing is an essential part of the heuristic that marries theoretical and empirical research, but experimental studies of resilience are lagging behind theory, management and policy requirements. Empirically, ecological resilience must be understood in terms of community dynamics and the potential for small shifts in environmental forcing to break the feedbacks that support resilience. Here, we integrate recent theory and empirical data to identify ways we might define and understand potential thresholds in the resilience of nature, and thus the potential for regime shifts, by focusing on the roles of strong and weak interactions, linkages in meta-communities, and positive feedbacks between these and environmental drivers. The challenge to theoretical and field ecologists is to make the shift from hindsight to a more predictive science that is able to assist in the implementation of ecosystem-based management. PMID- 19553255 TI - Separate and combined effects of nutrition during juvenile and sexual development on female life-history trajectories: the thrifty phenotype in a cockroach. AB - We have yet to understand fully how conditions during different periods of development interact to influence life-history structure. Can the negative effects of poor juvenile nutrition be overcome by a good adult diet, or are life history strategies set by early experience? Here, we tested the influence and interaction of different nutritional quality during juvenile and sexual development on female resource allocation physiology, life history and courtship behaviour in the cockroach, Nauphoeta cinerea. Nymphs were raised on either a good-quality or poor-quality diet. After adult eclosion, females were either switched to the opposite diet or remained on their original diet. We assessed mating behaviour and lifetime reproductive success for half of the females from each treatment. We evaluated reproductive investment, somatic investment and resource reallocation from reproduction to the soma via oocyte apoptosis in the remaining females. We found that poor juvenile conditions resulted in a fat phenotype with slow juvenile growth and short reproductive lifespan that could not be retrieved with a change in diet. Good juvenile conditions resulted in the converse, but again fixed, phenotype in adulthood. Thus, juvenile nutrition sets adult patterns of resource allocation. PMID- 19553256 TI - The rise and fall of celiac disease in the United States. AB - Because celiac disease is greatly under-diagnosed in the United States, a common assumption is that U.S. doctors and researchers always have considered the condition extremely rare. However, the disorder captured widespread medical attention at the beginning of the twentieth century. Luther Emmett Holt, a leading pediatrician, encouraged three other doctors to investigate the condition. Two helped to associate celiac disease with elite medical institutions. The third linked it to the marketing efforts of the United Fruit Company. Interest in celiac declined after 1965, partly as a result of the decreased concern with nutrition and nutritional disorders. PMID- 19553257 TI - Rapporteurs' report: workshop on ethical issues in diagnostic radiology. AB - This paper presents the summary reports of the session rapporteurs at the Workshop on Ethical Issues in Diagnostic Radiology. The summaries reflect the extent to which the topics discussed are well reflected in the papers presented in this proceedings. PMID- 19553258 TI - Harnessing the information contained within genome-wide association studies to improve individual prediction of complex disease risk. AB - The current paradigm within genetic diagnostics is to test individuals only at loci known to affect risk of complex disease-yet the technology exists to genotype an individual at thousands of loci across the genome. We investigated whether information from genome-wide association studies could be harnessed to improve discrimination of complex disease affection status. We employed genome wide data from the Wellcome Trust Case Control Consortium to test this hypothesis. Each disease cohort together with the same set of controls were split into two samples-a 'Training Set', where thousands of SNPs that might predispose to disease risk were identified and a 'Prediction Set', where the discriminatory ability of these SNPs was assessed. Genome-wide scores consisting of, for example, the total number of risk alleles an individual carries was calculated for each individual in the prediction set. Case-control status was regressed on this score and the area under the receiver operator characteristic curve (AUC) estimated. In most cases, a liberal inclusion of SNPs in the genome-wide score improved AUC compared with a more stringent selection of top SNPs, but did not perform as well as selection based upon established variants. The addition of genome-wide scores to known variant information produced only a limited increase in discriminative accuracy but was most effective for bipolar disorder, coronary heart disease and type II diabetes. We conclude that this small increase in discriminative accuracy is unlikely to be of diagnostic or predictive utility at the present time. PMID- 19553260 TI - Chronic wound sepsis due to retained vacuum assisted closure foam. PMID- 19553261 TI - Intravenous fluids in adults undergoing surgery. PMID- 19553259 TI - Common body mass index-associated variants confer risk of extreme obesity. AB - To investigate the genetic architecture of severe obesity, we performed a genome wide association study of 775 cases and 3197 unascertained controls at approximately 550,000 markers across the autosomal genome. We found convincing association to the previously described locus including the FTO gene. We also found evidence of association at a further six of 12 other loci previously reported to influence body mass index (BMI) in the general population and one of three associations to severe childhood and adult obesity and that cases have a higher proportion of risk-conferring alleles than controls. We found no evidence of homozygosity at any locus due to identity-by-descent associating with phenotype which would be indicative of rare, penetrant alleles, nor was there excess genome-wide homozygosity in cases relative to controls. Our results suggest that variants influencing BMI also contribute to severe obesity, a condition at the extreme of the phenotypic spectrum rather than a distinct condition. PMID- 19553262 TI - Canadian woman seeks class action suit over breast cancer tests. PMID- 19553263 TI - New network of weather stations in Africa aims to improve food security and health. PMID- 19553264 TI - Audit finds big improvement in treatment of myocardial infarction. PMID- 19553266 TI - Screening for abdominal aortic aneurysm. PMID- 19553268 TI - A case of secondary amenorrhoea. PMID- 19553267 TI - Analysis of cost effectiveness of screening Danish men aged 65 for abdominal aortic aneurysm. AB - OBJECTIVE: To assess the cost effectiveness of screening men aged 65 for abdominal aortic aneurysm. DESIGN: Cost effectiveness analysis based on a probabilistic, enhanced economic decision analytical model from screening to death. POPULATION AND SETTING: Hypothetical population of men aged 65 invited (or not invited) for ultrasound screening in the Danish healthcare system. DATA SOURCES: Published results from randomised trials and observational epidemiological studies retrieved from electronic bibliographic databases, and supplementary data obtained from the Danish Vascular Registry. DATA SYNTHESIS: A hybrid decision tree and Markov model was developed to simulate the short term and long term effects of screening for abdominal aortic aneurysm compared with no systematic screening on clinical and cost effectiveness outcomes. Probabilistic sensitivity analyses using Monte Carlo simulation were carried out. Results were presented in a cost effectiveness acceptability curve, an expected value of perfect information curve, and a curve showing the expected (net) number of avoided deaths from abdominal aortic aneurysm over time after the introduction of screening. The model was validated by calibrating base case health outcomes and expected activity levels against evidence from the recent Cochrane review of screening for abdominal aortic aneurysm. RESULTS: The estimated costs per quality adjusted life year (QALY) gained discounted at 3% per year over a lifetime for costs and QALYs was pound43 485 (euro54,852; $71,160). At a willingness to pay threshold of pound30,000 the probability of screening for abdominal aortic aneurysm being cost effective was less than 30%. One way sensitivity analyses showed the incremental cost effectiveness ratio varying from pound32,640 to pound66,001 per QALY. CONCLUSION: Screening for abdominal aortic aneurysm does not seem to be cost effective. Further research is needed on long term quality of life outcomes and costs. PMID- 19553272 TI - A woman with tuberous sclerosis and acute onset right sided abdominal pain. PMID- 19553274 TI - Legalising drugs would be "an historic mistake," UN report says. PMID- 19553269 TI - Screening men for abdominal aortic aneurysm: 10 year mortality and cost effectiveness results from the randomised Multicentre Aneurysm Screening Study. AB - OBJECTIVES: To assess whether the mortality benefit from screening men aged 65-74 for abdominal aortic aneurysm decreases over time, and to estimate the long term cost effectiveness of screening. DESIGN: Randomised trial with 10 years of follow up. SETTING: Four centres in the UK. Screening and surveillance was delivered mainly in primary care settings, with follow-up and surgery offered in hospitals. PARTICIPANTS: Population based sample of 67 770 men aged 65-74. INTERVENTIONS: Participants were individually allocated to invitation to ultrasound screening (invited group) or to a control group not offered screening. Patients with an abdominal aortic aneurysm detected at screening underwent surveillance and were offered surgery if they met predefined criteria. MAIN OUTCOME MEASURES: Mortality and costs related to abdominal aortic aneurysm, and cost per life year gained. RESULTS: Over 10 years 155 deaths related to abdominal aortic aneurysm (absolute risk 0.46%) occurred in the invited group and 296 (0.87%) in the control group (relative risk reduction 48%, 95% confidence interval 37% to 57%). The degree of benefit seen in earlier years of follow-up was maintained in later years. Based on the 10 year trial data, the incremental cost per man invited to screening was pound100 (95% confidence interval pound82 to pound118), leading to an incremental cost effectiveness ratio of pound7600 ( pound5100 to pound13,000) per life year gained. However, the incidence of ruptured abdominal aortic aneurysms in those originally screened as normal increased noticeably after eight years. CONCLUSIONS: The mortality benefit of screening men aged 65-74 for abdominal aortic aneurysm is maintained up to 10 years and cost effectiveness becomes more favourable over time. To maximise the benefit from a screening programme, emphasis should be placed on achieving a high initial rate of attendance and good adherence to clinical follow-up, preventing delays in undertaking surgery, and maintaining a low operative mortality after elective surgery. On the basis of current evidence, rescreening of those originally screened as normal is not justified. Trial registration Current Controlled Trials ISRCTN37381646. PMID- 19553275 TI - Parkinson's disease, DBS and suicide: a role for serotonin? PMID- 19553277 TI - Flavonoids inhibit the genotoxicity of hydrogen peroxide (H(2)O(2)) and of the food mutagen 2-amino-3-methylimadazo[4,5-f]-quinoline (IQ) in lymphocytes from patients with inflammatory bowel disease (IBD). AB - Inflammatory bowel disease (IBD) including Crohn's disease (CD) and ulcerative colitis (UC) is a chronic inflammatory gastrointestinal autoimmune condition with an inappropriate immune response. We investigated DNA damage induced in vitro in lymphocytes from IBD patients caused by oxidative stress through H(2)O(2) and 2 amino-3-methylimidazo[4,5-f]quinoline (IQ) and whether the plant flavonoids, quercetin and epicatechin, found in fruits, tea and soybeans could effectively reduce such stress. Lymphocytes from IBD patients and healthy volunteers were treated with 50 microg/ml H(2)O(2) or IQ in the presence of quercetin (0-250 microg/ml) or epicatechin (0-100 microg/ml). Flavonoid supplementation (250 microM quercetin or 100 microM epicatechin) caused an overall significant decrease of induced DNA damage resulting in a 48.6% (P < 0.001) reduction of H(2)O(2)-induced and a 43% (P < 0.001) reduction of IQ-induced DNA damage within the patient groups; for the control groups, reductions in DNA damage were 35.2 and 57.1%, respectively (both, P < 0.001). There was less induced DNA damage within lymphocytes from UC patients compared to CD patients for both series of experiments (H(2)O(2) and quercetin, IQ and epicatechin). In conclusion, flavonoids dramatically reduced oxidative stress in vitro in lymphocytes from IBD patients and healthy individuals. Thus, flavonoids could be very effective in the treatment of oxidative stress and encouraged in the diet of IBD patients. PMID- 19553278 TI - Measurements come in all shapes and sizes. PMID- 19553279 TI - Toll-like receptor agonist induced changes in clonal rat BRIN-BD11 beta-cell insulin secretion and signal transduction. AB - Evidence for involvement of toll-like receptors (TLRs) (e.g. TLR4 and TLR2, whose agonists include lipopolysaccharides (LPS) and saturated fatty acids) in altered patterns of signalling in adipose, liver and muscle from animal models of insulin resistance and obesity has been published. We have now extended this area of research and have determined the effects of LPS on cell viability, insulin secretion, insulin signalling and metabolism in a clonal beta-cell line. BRIN BD11 beta-cells were treated for 24 h with increasing concentrations of LPS. Chronic (24 h) and acute (20 min) insulin secretion, insulin content and parameters of cell metabolism and insulin signalling were determined. Incubation of BRIN-BD11 cells for 24 h in the presence of increasing concentrations of the TLR4 ligand LPS significantly decreased chronic (24 h) insulin secretion from 1.09+/-0.19 to 0.76+/-0.18 microg insulin/mg protein in the presence of 100 ng/ml LPS (P<0.05). There was no change in acute (20 min) stimulated insulin secretion or insulin content. Cell metabolism was not changed. Insulin receptor-beta (IR beta) expression levels were increased significantly from 1+/-0.52 to 8.6+/-1.83 units (P<0.01), whereas calcineurin activity and Akt phosphorylation were significantly (P<0.01 and P<0.05 respectively) reduced in response to 24 h incubation in the presence of LPS. There was no change in IR substrate-1 protein expression or phosphorylation after 24 h. Further incubation for 24 h in the absence of LPS resulted in the recovery of chronic insulin secretion. The negative beta-cell effects of LPS may contribute to hyperglycaemia in vivo. PMID- 19553280 TI - Short- and long-term effects of maternal nicotine exposure during lactation on body adiposity, lipid profile, and thyroid function of rat offspring. AB - Epidemiological studies show a higher prevalence of obesity in children from smoking mothers and smoking may affect human thyroid function. To evaluate the mechanism of smoking as an imprinting factor for these dysfunctions, we evaluated the programming effects of maternal nicotine (NIC) exposure during lactation. Two days after birth, osmotic minipumps were implanted in lactating rats, divided into: NIC (6 mg/kg per day s.c.) for 14 days; Control - saline. All the significant data were P<0.05 or less. Body weight was increased from 165 days old onwards in NIC offspring. Both during exposure (at 15 days old) and in adulthood (180 days old), NIC group showed higher total fat (27 and 33%). In addition, NIC offspring presented increased visceral fat and total body protein. Lipid profile was not changed in adulthood. Leptinemia was higher at 15 and 180 days old (36 and 113%), with no changes in food intake. Concerning the thyroid status, the 15 days-old NIC offspring showed lower serum-free tri-iodothyronine (FT(3)) and thyroxine (FT(4)) with higher TSH. The 180-days-old NIC offspring exhibited lower TSH, FT(3), and FT(4)). In both periods, liver type 1 deiodinase was lower (26 and 55%). We evidenced that NIC imprints a neonatal thyroid dysfunction and programs for a higher adiposity, hyperleptinemia, and secondary hypothyroidism in adulthood. Our study identifies lactation as a critical period to NIC programming for obesity, with hypothyroidism being a possible contributing factor. PMID- 19553281 TI - Identification, sequencing, and cellular localization of hepcidin in guinea pig (Cavia porcellus). AB - Hepcidin, a cysteine-rich peptide hormone with antimicrobial and iron-regulatory activity, plays a central role in regulating iron metabolism during inflammation, hypoxia, iron deficiency, and iron overload. The aim of this study was to isolate and sequence the guinea pig hepcidin gene and show peptide's tissue distribution to identify the guinea pig as good animal model to study the regulation and function of hepcidin. The guinea pig hepcidin cDNA contains a 252 bp open reading frame encoding for an 83 amino acid protein with eight highly conserved cysteine residues. Phylogenetic analyses showed that guinea pig hepcidin was more related to human and chimpanzee than to rodents like mouse or rat. RT-PCR studies revealed that hepcidin mRNA was most abundant in liver, less ample in pancreas, heart, and kidney and not detectable in lung and biliary system. Western blot analyses showed a distinct immunoreactive band of approximately 8 kDa, consistent with the predicted size of prohepcidin, and revealed that guinea pig hepcidin protein is synthesized predominantly in the liver, and with lower expression in kidney, heart, and pancreas. Immunohistochemical studies showed hepcidin predominantly at the basolateral membrane domain of hepatocytes in periportal regions. In pancreas, hepcidin immunoreactivity was confined to endocrine islets of Langerhans, while hepcidin was seen in tubules, but not in the glomeruli in the kidney. Our data identify guinea pig as a convenient model organism to study the role of hepcidin, given the remarkable sequence similarity and tissue distribution pattern largely identical to human. PMID- 19553283 TI - Retraction: high, but not low, molecular weight preparations of hyaluronic acid protect asthmatics against challenge-induced bronchoconstriction. PMID- 19553282 TI - Ultrasound feedback and motivational interviewing targeting smoking cessation in the second and third trimesters of pregnancy. AB - INTRODUCTION: Cigarette smoking during pregnancy is associated with poor maternal and child health outcomes. Effective interventions to increase smoking cessation rates are needed particularly for pregnant women unable to quit in their first trimester. Real-time ultrasound feedback focused on potential effects of smoking on the fetus may be an effective treatment adjunct, improving smoking outcomes. METHODS: A prospective randomized trial was conducted to evaluate the efficacy of a smoking cessation intervention consisting of personalized feedback during ultrasound plus motivational interviewing-based counseling sessions. Pregnant smokers (N = 360) between 16 and 26 weeks of gestation were randomly assigned to one of three groups: Best Practice (BP) only, Best Practice plus ultrasound feedback (BP+US), or Motivational Interviewing-based counseling plus ultrasound feedback (MI+US). Assessments were conducted at baseline and end of pregnancy (EOP). RESULTS: Analyses of cotinine-verified self-reported smoking status at EOP indicated that 10.8% of the BP group was not smoking at EOP; 14.2% in the BP+US condition and 18.3% who received MI+US were abstinent, but differences were not statistically significant. Intervention effects were found conditional upon level of baseline smoking, however. Nearly 34% of light smokers (< or =10 cigarettes/day) in the MI+US condition were abstinent at EOP, followed by 25.8% and 15.6% in the BP+US and BP conditions, respectively. Heavy smokers (>10 cigarettes/day) were notably unaffected by the intervention. DISCUSSION: Future research should confirm benefit of motivational interviewing plus ultrasound feedback for pregnant light smokers and explore mechanisms of action. Innovative interventions for pregnant women smoking at high levels are sorely needed. PMID- 19553284 TI - Two distinct types of mouse melanocyte: differential signaling requirement for the maintenance of non-cutaneous and dermal versus epidermal melanocytes. AB - Unlike the thoroughly investigated melanocyte population in the hair follicle of the epidermis, the growth and differentiation requirements of the melanocytes in the eye, harderian gland and inner ear - the so-called non-cutaneous melanocytes remain unclear. In this study, we investigated the in vitro and in vivo effects of the factors that regulate melanocyte development on the stem cells or the precursors of these non-cutaneous melanocytes. In general, a reduction in KIT receptor tyrosine kinase signaling leads to disordered melanocyte development. However, melanocytes in the eye, ear and harderian gland were revealed to be less sensitive to KIT signaling than cutaneous melanocytes. Instead, melanocytes in the eye and harderian gland were stimulated more effectively by endothelin 3 (ET3) or hepatocyte growth factor (HGF) signals than by KIT signaling, and the precursors of these melanocytes expressed the lowest amount of KIT. The growth and differentiation of these non-cutaneous melanocytes were specifically inhibited by antagonists for ET3 and HGF. In transgenic mice induced to express ET3 or HGF in their skin and epithelial tissues from human cytokeratin 14 promoters, the survival and differentiation of non-cutaneous and dermal melanocytes, but not epidermal melanocytes, were enhanced, apparently irrespective of KIT signaling. These results provide a molecular basis for the clear discrimination between non-cutaneous or dermal melanocytes and epidermal melanocytes, a difference that might be important in the pathogenesis of melanocyte-related diseases and melanomas. PMID- 19553286 TI - Beta-catenin controls differentiation of the retinal pigment epithelium in the mouse optic cup by regulating Mitf and Otx2 expression. AB - The retinal pigment epithelium (RPE) consists of a monolayer of cuboidal, pigmented cells that is located between the retina and the choroid. The RPE is vital for growth and function of the vertebrate eye and improper development results in congenital defects, such as microphthalmia or anophthalmia, or a change of cell fate into neural retina called transdifferentiation. The transcription factors microphthalmia-associated transcription factor (Mitf) and orthodenticle homolog 2 (Otx2) are crucial for RPE development and function; however, very little is known about their regulation. Here, by using a Wnt responsive reporter, we show that the Wnt/beta-catenin pathway is activated in the differentiating mouse RPE. Cre-mediated, RPE-specific disruption of beta catenin after the onset of RPE specification causes severe defects, resulting in microphthalmia with coloboma, disturbed lamination, and mislocalization of adherens junction proteins. Upon beta-catenin deletion, the RPE transforms into a multilayered tissue in which the expression of Mitf and Otx2 is downregulated, while retina-specific gene expression is induced, which results in the transdifferentiation of RPE into retina. Chromatin immunoprecipitation (ChIP) and luciferase assays indicate that beta-catenin binds near to and activates potential TCF/LEF sites in the Mitf and Otx2 enhancers. We conclude that Wnt/beta catenin signaling is required for differentiation of the RPE by directly regulating the expression of Mitf and Otx2. Our study is the first to show that an extracellular signaling pathway directly regulates the expression of RPE specific genes such as Mitf and Otx2, and elucidates a new role for the Wnt/beta catenin pathway in organ formation and development. PMID- 19553287 TI - Associations of maternal lifetime trauma and perinatal traumatic stress symptoms with infant cardiorespiratory reactivity to psychological challenge. AB - OBJECTIVE: To examine associations of maternal lifetime trauma and related psychological symptoms in the perinatal period with infant cardiorespiratory reactivity and behavioral distress in response to a laboratory stressor, using a novel advanced system recently adapted for infants. METHODS: Participants were mothers and their 6-month-old infants. Assessments included mothers' self reported lifetime exposure to trauma, perinatal traumatic stress, and current symptoms of posttraumatic stress disorder (PTSD) and depression. Through the use of a noninvasive respiratory inductance plethysmography device, heart rate and indices of respiratory volume, timing, and thoracoabdominal coordination were recorded continuously in 23 infants during the Still-Face Paradigm, a videotaped mother-infant dyadic assessment that included baseline, stressor, and recovery phases. Infant behavioral distress during the procedure was also assessed. RESULTS: Infants of mothers with low exposure to trauma and perinatal traumatic stress showed expected increases in behavioral distress and cardiorespiratory activation from baseline to stressor and decreases in these parameters from stressor to recovery. Infants of mothers exposed to multiple traumas and with elevated perinatal traumatic stress showed similar patterns of activation from baseline to stressor but failed to show decreases during recovery. These patterns were maintained after controlling for current maternal PTSD and depressive symptoms. CONCLUSIONS: Maternal lifetime trauma exposure and traumatic stress during the perinatal period were associated with disrupted infant cardiorespiratory regulation and behavioral distress during a stressor protocol. These results support the concept of perinatal programming and its potential role in physical and mental health outcomes. PMID- 19553285 TI - Lunatic fringe promotes the lateral inhibition of neurogenesis. AB - Previous studies have identified roles of the modulation of Notch activation by Fringe homologues in boundary formation and in regulating the differentiation of vertebrate thymocytes and Drosophila glial cells. We have investigated the role of Lunatic fringe (Lfng) expression during neurogenesis in the vertebrate neural tube. We find that in the zebrafish hindbrain, Lfng is expressed by progenitors in neurogenic regions and downregulated in cells that have initiated neuronal differentiation. Lfng is required cell autonomously in neural epithelial cells to limit the amount of neurogenesis and to maintain progenitors. By contrast, Lfng is not required for the role of Notch in interneuronal fate choice, which we show is mediated by Notch1a. The expression of Lfng does not require Notch activity, but rather is regulated downstream of proneural genes that are widely expressed by neural progenitors. These findings suggest that Lfng acts in a feedback loop downstream of proneural genes, which, by promoting Notch activation, maintains the sensitivity of progenitors to lateral inhibition and thus limits further proneural upregulation. PMID- 19553288 TI - Hostility and fasting glucose in African American women. AB - OBJECTIVE: To examine whether the relationship of hostility (HOST) to fasting glucose indices is moderated by sex and race. HOST has been associated with abnormalities in glucose metabolism. Prior studies suggested that this association may be more prevalent in women and in African American (AA) individuals. METHODS: A total of 565 healthy AA and white (W) men and women (mean age = 33 +/- 6 years) were assessed. HOST was measured by the 27-item version of the Cook Medley HOST Scale. The moderating effects of sex and race were evaluated for the associations of HOST to fasting glucose, insulin, and insulin sensitivity (HOMA-IR). RESULTS: Analysis showed a moderating effect of sex and race on the association of HOST to fasting glucose (p = .03), but not for insulin (p = .12). Analysis of HOMA-IR revealed a trend (p = .06) for the interaction. Stratified analyses by race and sex revealed a positive association between HOST and fasting glucose only in AA women, which remained significant after controlling for age and body mass index. CONCLUSION: A relationship between HOST and fasting glucose was evident in AA women only, a group that has twice the risk of developing Type 2 diabetes compared with W women. Further studies are needed to elucidate the mechanisms by which HOST may affect glucose metabolism in AA women. PMID- 19553291 TI - Gefitinib treatment in hormone-resistant and hormone receptor-negative advanced breast cancer. AB - BACKGROUND: Acquired and de novo endocrine resistance in breast cancer (BC) may be associated with overexpression of epidermal growth factor receptor (EGFR). Gefinitib is an orally active selective EGFR inhibitor which might benefit advanced breast cancer (ABC) patients either with acquired hormone resistance or with hormone receptor (HR)-negative tumors. PATIENTS AND METHODS: A two-arm multicenter phase II trial of oral gefitinib 500 mg/day was planned in two groups of 45 patients with ABC for whom chemotherapy was not currently indicated. Group 1 had hormone-resistant BC defined as HR-positive BC with progression after treatment with tamoxifen and an aromatase inhibitor. Group 2 had HR-negative BC. Tumor response was assessed every 8 weeks. The primary end point was the clinical benefit rate (CBR). RESULTS: Forty patients with hormone-resistant BC had a CBR of 0%. Two of 25 HR-negative BC patients showed stable disease (less than a 50% reduction and less than a 25% increase in the sum of the products of two perpendicular diameters of all measured lesions and the appearance of no new lesions) at 24 weeks resulting in a CBR of 7.7% (95% CI 0.9% to 25.1%). Enrollment ceased due to the low CBR. Toxicity resulted in treatment interruption (46%), dose reduction (20%) and withdrawal (11%) of patients. CONCLUSION: At a dose of 500 mg/day, gefitinib monotherapy resulted in a low CBR and no tumor response was identified. PMID- 19553289 TI - Social stress desensitizes lymphocytes to regulation by endogenous glucocorticoids: insights from in vivo cell trafficking dynamics in rhesus macaques. AB - OBJECTIVE: To determine whether chronic social stress can desensitize leukocytes to normal physiologic regulation by endogenous glucocorticoids. METHODS: We analyzed the longitudinal relationship between plasma cortisol levels and peripheral blood lymphocyte counts over 16 monthly assessments in 18 rhesus macaques randomized to recurrent social encounters with a stable set of conspecifics or continually varying social partners (unstable socialization). RESULTS: Animals socialized under stable conditions showed the expected inverse relationship between plasma cortisol concentrations and circulating lymphocyte frequencies. That relationship was significantly attenuated in animals subject to unstable social conditions. Differences in leukocyte redistributional sensitivity to endogenous glucocorticoids emerged within the first week of differential socialization, persisted throughout the 60-week study period, and were correlated with other measures of glucocorticoid desensitization (blunted hypothalamic pituitary-adrenal (HPA) axis response to acute stress and redistributional response to dexamethasone challenge). Effects of unstable social conditions on leukocyte sensitivity to cortisol regulation were not related to physical aggression. CONCLUSION: Chronic social stress can impair normal physiologic regulation of leukocyte function by the HPA axis in ways that may contribute to the increased physical health risks associated with social adversity. PMID- 19553290 TI - Does anxiety affect risk of dementia? Findings from the Caerphilly Prospective Study. AB - OBJECTIVE: To examine the association of anxiety with incident dementia and cognitive impairment not dementia (CIND). METHODS: We conducted a prospective study of men aged 48 to 67 years at baseline anxiety assessment; we measured cognition 17 years later. We studied 1481 men who were either eligible for examination or were known to have dementia. Trait Anxiety was assessed using the Spielberger State Trait Anxiety Inventory. Psychological distress was assessed using the 30-item general health questionnaire. Cognitive screening was followed by a clinical examination. Medical notes and death certificates of those not seen were also examined. Outcomes were CIND and Diagnostic and Statistical Manual of Mental Disorders, 4th Edition (DSM-IV) dementia. RESULTS: Of 1160 men who were cognitively screened, 174 cases of CIND and 69 cases of dementia were identified. A further 21 cases of dementia were identified from medical records. After adjustment for age, vascular risk factors and premorbid cognitive function associations with higher anxiety (31st-95th centile) were for CIND odds ratio (OR) 2.31 (95% Confidence Interval (CI) = 1.20-4.44) and for dementia OR 2.37 (95% CI = 0.98-5.71). These associations were slightly stronger for nonvascular (OR = 2.45; 95% CI = 1.28-4.68) than for vascular impairment (OR = 1.94; 95% CI = 0.77-4.89). Analyses of change in cognitive performance, assessed by the Cambridge Cognitive Examination of the Elderly subscales found some evidence for decline in learning memory with higher anxiety score (b(age adj) = -0.291 ( 0.551, -0.032), but not for any other subscale. CONCLUSIONS: Anxiety is a risk factor for CIND and dementia. The extent to which the association is independent of depression and whether or not it is causal requires further study. PMID- 19553292 TI - Glycemic index, glycemic load and renal cell carcinoma risk. AB - BACKGROUND: The risk of renal cell carcinoma (RCC) has been related to refined cereals and starchy foods, but the association has not been studied in terms of glycemic index (GI) and glycemic load (GL). To provide information on this issue, we analyzed data from an Italian multicentric case-control study. MATERIALS AND METHODS: Cases were 767 patients with histologically confirmed, incident RCC. Controls were 1534 subjects admitted to the same hospitals as cases for a wide spectrum of acute, non-neoplastic conditions, unrelated to known risk factors for RCC. Information on dietary habits was derived through a food-frequency questionnaire. Multivariate odds ratios (ORs) and 95% confidence intervals (CIs) for GI and GL intake were adjusted for major relevant covariates. RESULTS: Compared with the lowest quintile, the ORs for the highest quintile were 1.43 (95% CI 1.05-1.95) for GI and 2.56 (95% CI 1.78-3.70) for GL, with significant trends in risk. Compared with the lowest quintile, the risk of RCC for all subsequent levels of GL was higher in never drinkers than in ever drinkers. CONCLUSIONS: We found direct relations between dietary levels of GI and GL and RCC risk. This can be related to mechanisms linked to insulin resistance and sensitivity. PMID- 19553293 TI - Can we reduce snack food intake? PMID- 19553294 TI - Physical activity attenuates the body mass index-increasing influence of genetic variation in the FTO gene. AB - BACKGROUND: Intronic variation in the FTO (fat mass and obesity-associated) gene has been unequivocally associated with increased body mass index (BMI; in kg/m(2)) and the risk of obesity in populations of different ethnicity. OBJECTIVE: We examined whether this robust genetic predisposition to obesity can be attenuated by being more physically active. DESIGN: The FTO variant rs1121980 was genotyped in 20,374 participants (39-79 y of age) from the European Prospective Investigation into Cancer and Nutrition-Norfolk Study, an ethnically homogeneous population-based cohort. Physical activity (PA) was assessed with a validated self-reported questionnaire. The interaction between rs1121980 and PA on BMI and waist circumference (WC) was examined by including the interaction term in mixed-effect models. RESULTS: We confirmed that the risk (T) allele of rs1121980 was significantly associated with BMI (0.31-unit increase per allele; P < 0.001) and WC (0.77-cm increase per allele; P < 0.001). The PA level attenuated the effect of rs1121980 on BMI and WC; ie, whereas in active individuals the risk allele increased BMI by 0.25 per allele, the increase in BMI was significantly (P for interaction = 0.004) more pronounced (76%) in inactive individuals (0.44 per risk allele). We observed similar effects for WC (P for interaction = 0.02): the risk allele increased WC by 1.04 cm per allele in inactive individuals but by only 0.64 cm in active individuals. CONCLUSIONS: Our results showed that PA attenuates the effect of the FTO rs1121980 genotype on BMI and WC. This observation has important public health implications because we showed that a genetic susceptibility to obesity induced by FTO variation can be overcome, at least in part, by adopting a physically active lifestyle. PMID- 19553295 TI - Lysine ingestion markedly attenuates the glucose response to ingested glucose without a change in insulin response. AB - BACKGROUND: Ingested proteins are known to stimulate a rise in insulin and glucagon concentrations. In our effort to explain this effect, we have begun to measure the effect of individual amino acids. OBJECTIVES: The objectives were to determine the effect of lysine ingestion on insulin and glucagon concentrations and whether the effect is moderated by glucose ingestion. DESIGN: Thirteen healthy subjects were studied on 4 occasions. Water, 25 g glucose, 1 mmol lysine/kg lean body mass, or lysine plus glucose was given on separate occasions at 0800 after a 12-h fast. Serum lysine, glucose, insulin, and glucagon were measured during a 2.5-h period. The amount of lysine provided was equivalent to that present in a 672-g (24-oz) steak. RESULTS: Lysine ingestion resulted in an approximately 3-fold increase in lysine concentration and in a small decrease in glucose concentration. When lysine was ingested with glucose, the 2.5-h glucose area response decreased by 44% (P < 0.02). Lysine alone increased the insulin area response modestly; the insulin increase when lysine was ingested with glucose was similar to that when only glucose was ingested. Lysine stimulated an increase in glucagon (P < 0.02), whereas glucose decreased glucagon. CONCLUSIONS: Lysine ingestion results in a small decrease in serum glucose and an increase in glucagon and insulin concentrations. Lysine ingested with glucose dramatically attenuated the glucose-stimulated glucose response, but there was no change in insulin response. Whether similar effects will be observed with more physiologic doses of lysine remains to be determined. PMID- 19553296 TI - Effect of zinc supplementation on morbidity and growth in hospital-born, low birth-weight infants. AB - BACKGROUND: Low-birth-weight infants may have impaired zinc status, but little is known about the effect of zinc supplementation. OBJECTIVE: The objective was to investigate the effect of daily zinc supplementation on morbidity and anthropometric status in hospital-born, low-birth-weight infants. DESIGN: In a double-blind, randomized, placebo-controlled trial, 2052 hospital-born term infants with a birth weight < or =2500 g were randomly assigned to receive zinc or placebo. The zinc group received elemental zinc: 5 mg/d for those infants between ages 2 wk and 6 mo and 10 mg/d for those infants aged >6 mo. All-cause hospitalizations, prevalence of diarrhea, acute lower respiratory tract infections, visits to health care providers, weights, and lengths were ascertained at 3, 6, 9, and 12 mo of age. RESULTS: The supplement was consumed for >85% of the follow-up period. Mean plasma zinc at 12 mo of age was higher in the zinc group (100.2 microg/dL) than in the control group (73.3 microg/dL) (difference in means: 26.9; 95% CI: 19.6, 34.2). The 24-h and 7-d prevalence of diarrhea and acute lower respiratory tract infections was similar at 3, 6, 9, and 12 mo. Care-seeking for illness was significantly lower in the zinc group (difference in proportions: -5.7; 95% CI: -9.9, -1.4; P < 0.05) at 9 mo. The numbers of hospitalizations, weights, and lengths were all similar at all 4 assessments. CONCLUSION: Hospital-born, term, low-birth-weight infants do not seem to benefit substantially from zinc supplementation that meets the Recommended Dietary Allowance for zinc in terms of morbidity or physical growth during infancy in this setting. This trial was registered at www.clinicaltrials.gov as NCT00272142. PMID- 19553297 TI - An obesogenic postnatal environment is more important than the fetal environment for the development of adult adiposity: a study of female twins. AB - BACKGROUND: A relation between birth weight and adult body composition has been reported in singleton populations, especially when more accurate measures of body composition, such as dual-energy X-ray absorptiometry (DXA) were used. It remains uncertain whether this is mediated by a direct effect of fetal nutrition, through factors in the shared environment, or through genetic factors. OBJECTIVE: The objective was to investigate the relation between birth weight and body composition with the use of a co-twin design. DESIGN: DXA measurements and birth weights were available for 2228 dizygotic and 842 monozygotic female twins aged between 18 and 80 y. Multivariate regression models were used to identify both individual specific relations and those mediated through the shared environment. RESULTS: Significant relations were found between birth weight and DXA measures for individuals. A 1-kg increase in birth weight was associated with a 1.72-kg increase in lean mass, a 0.25-kg increase in fat mass, and a 0.05-unit increase in the lean:fat mass ratio. Within twin pairs, the analysis showed that associations between birth weight and absolute levels of lean and fat mass were mediated through individual-specific effects, whereas the relation between birth weight and the proportion of lean to fat mass was mediated purely through factors common to twin pairs. CONCLUSIONS: A higher birth weight is associated with a higher proportion of lean to fat mass as adults. However, these analyses suggest that this association is not determined by individual specific factors in utero (eg, fetal nutrition) but through factors in the shared common environment of the twins. PMID- 19553299 TI - Poor vitamin C status is associated with increased carotid intima-media thickness, decreased microvascular function, and delayed myocardial repolarization in young patients with type 1 diabetes. AB - BACKGROUND: Vascular endothelial dysfunction, accelerated thickening of arterial intima, and changes in ventricular repolarization contribute to increased cardiovascular morbidity in type 1 diabetes (T1D). Although vitamin C has important antioxidant functions and increased oxidative stress is a central mechanism of cardiovascular dysfunction in T1D, the relation between vitamin C and the cardiovascular system in young diabetic patients has not been investigated. OBJECTIVE: In a cohort of young patients with T1D, we investigated the relation of plasma concentrations of vitamin C with indexes of vascular function and structure and duration of the QT interval corrected for heart rate (QT(c)). DESIGN: Carotid artery intima-media thickness, cutaneous microvascular function, and duration of the QT(c) interval were measured in 59 patients (mean age: 17 y; range: 10-22 y) with T1D (diabetes duration: 3-20 y). Plasma vitamin C was analyzed by HPLC with coulometric detection. RESULTS: Carotid artery intima media thickness and duration of the QT(c) interval were higher in patients in the lowest tertile of vitamin C than in those in the highest tertile (P < 0.05 for both). The cutaneous microvascular response to acetylcholine was lower (P = 0.003) in the lowest tertile group than in the highest tertile group, but the response to sodium nitroprusside was not significantly different between these 2 groups. All differences remained significant after adjustment for age, sex, diabetes duration, body mass index, and glycated hemoglobin. CONCLUSIONS: In this relatively small-scale cross-sectional study of young patients with T1D, lower plasma concentrations of vitamin C seem to be associated with adverse changes in the microcirculation, peripheral arteries, and ventricular repolarization. Large scale prospective studies are needed to confirm these results and to clarify the underlying mechanisms. PMID- 19553298 TI - Dietary fish and meat intake and dementia in Latin America, China, and India: a 10/66 Dementia Research Group population-based study. AB - BACKGROUND: Evidence of an association between fish and meat consumption and risk of dementia is inconsistent and nonexistent in populations in developing countries. OBJECTIVE: The objective was to investigate associations between fish and meat consumption with dementia in low- and middle-income countries. DESIGN: One-phase cross-sectional surveys were conducted in all residents aged > or =65 y in 11 catchment areas in China, India, Cuba, the Dominican Republic, Venezuela, Mexico, and Peru. A total of 14,960 residents were assessed by using the 10/66 standardized protocol, which includes face-to-face interviews for dietary habits and a cross-culturally validated dementia diagnosis. RESULTS: Dietary intakes and the prevalence of dementia varied between sites. We combined site-specific Poisson regression prevalence ratios (PRs) for the association between fish and meat consumption and dementia in 2 fixed-effect model meta-analyses adjusted for sociodemographic and health characteristics and fish and meat consumption as appropriate. We found a dose-dependent inverse association between fish consumption and dementia (PR: 0.81; 95% CI: 0.72, 0.91) that was consistent across all sites except India and a less-consistent, dose-dependent, direct association between meat consumption and prevalence of dementia (PR: 1.19; 95% CI: 1.07, 1.31). CONCLUSIONS: Our results extend findings on the associations of fish and meat consumption with dementia risk to populations in low- and middle income countries and are consistent with mechanistic data on the neuroprotective actions of omega-3 (n-3) long-chain polyunsaturated fatty acids commonly found in fish. The inverse association between fish and prevalent dementia is unlikely to result from poorer dietary habits among demented individuals (reverse causality) because meat consumption was higher in those with a diagnosis of dementia. PMID- 19553301 TI - Fish consumption and markers of colorectal cancer risk: a multicenter randomized controlled trial. AB - BACKGROUND: Diet is a major factor in the etiology of colorectal cancer, with high fish consumption possibly decreasing colorectal cancer risk, as was shown in several observational studies. To date, no intervention trials have examined the possible beneficial effects of fish intake on colorectal cancer risk. OBJECTIVE: The objective was to investigate the effects of a 6-mo intervention with oil-rich or lean fish on apoptosis and mitosis within the colonic crypt. DESIGN: In a multicenter, randomized, controlled intervention trial, patients with colorectal polyps, inactive ulcerative colitis, or no macroscopic signs of disease were recruited (n = 242) and randomly allocated to receive dietary advice plus either 300 g oil-rich fish (salmon) per week (n = 82), 300 g lean fish (cod) per week (n = 78), or only dietary advice (DA) (n = 82). Apoptosis and mitosis were measured in colonic biopsy samples collected before and after intervention (n = 213). RESULTS: The total number of apoptotic cells per crypt did not increase in the salmon or cod group: -0.10 (95% CI: -0.36, 0.16) and -0.06 (95% CI: -0.32, 0.20), respectively, compared with the DA group. The total number of mitotic cells per crypt decreased nonsignificantly in the salmon group (-0.87; 95% CI: -2.41, 0.68) and in the cod group (-1.04; 95% CI: -2.62, 0.53) compared with the DA group. Furthermore, the distribution of mitosis within the crypt did not significantly change in either group. CONCLUSION: An increase in the consumption of either oil rich or lean fish to 2 portions weekly over 6 mo does not markedly change apoptotic and mitotic rates in the colonic mucosa. This trial was registered at www.clinicaltrials.gov as NCT00145015. PMID- 19553302 TI - Diabetes risk: antioxidants or lifestyle? PMID- 19553300 TI - Growth faltering due to breastfeeding cessation in uninfected children born to HIV-infected mothers in Zambia. AB - BACKGROUND: The effect of breastfeeding on growth in HIV-exposed infants is not well described. OBJECTIVE: The objective was to evaluate the effect of early breastfeeding cessation on growth. DESIGN: In a trial conducted in Lusaka, Zambia, HIV-infected mothers were randomly assigned to exclusive breastfeeding for 4 mo followed by rapid weaning to replacement foods or exclusive breastfeeding for 6 mo followed by introduction of complementary foods and continued breastfeeding for a duration of the mother's choice. Weight-for-age z score (WAZ), length-for-age z score (LAZ), and weight-for-length z score (WLZ) and the self-reported breastfeeding practices of 593 HIV-uninfected singletons were analyzed. Generalized estimating equations were used to adjust for confounders. RESULTS: WAZ scores declined precipitously between 4.5 and 15 mo. The decline was slower in the breastfed infants. At 9, 12, and 15 mo, mean WAZs were, respectively, -0.74, -0.92, and -1.06 in infants who were reportedly breastfed and were -1.07, -1.20, and -1.31 in the weaned infants (P = 0.003, 0.007, and 0.02, respectively). No differences were observed past 15 mo. Breastfeeding practice was not associated with LAZ, which declined from -0.98 to 2.24 from 1 to 24 mo. After adjustment for birth weight, maternal viral load, body mass index, education, season, and marital and socioeconomic status, not breastfeeding was associated with a 0.28 decline in WAZ between 4.5 and 15 mo (P < 0.0001). During the rainy season, not breastfeeding was associated with a larger WAZ decline (0.33) than during the dry season (0.22; P for interaction = 0.02). CONCLUSIONS: Early growth is compromised in uninfected children born to HIV-infected Zambian mothers. Continued breastfeeding partially mitigates this effect through 15 mo. Nutritional interventions to complement breastfeeding after 6 mo are urgently needed. This trial was registered at clinicaltrials.gov as NCT00310726. PMID- 19553303 TI - Analysis of the kinetics of transcription and replication of the rotavirus genome by RNA interference. AB - Rotaviruses have a genome composed of 11 segments of double-stranded RNA (dsRNA) surrounded by three protein layers. The virus contains an RNA-dependent RNA polymerase that synthesizes RNA transcripts corresponding to all segments of the viral genome. These transcripts direct the synthesis of the viral proteins and also serve as templates for the synthesis of the complementary strand to form the dsRNA genome. In this work, we analyzed the kinetics of transcription and replication of the viral genome throughout the replication cycle of the virus using quantitative reverse transcription-PCR. The role of the proteins that form double-layered particles ([DLPs] VP1, VP2, VP3, and VP6) in replication and transcription of the viral genome was analyzed by silencing their expression in rotavirus-infected cells. All of them were shown to be essential for the replication of the dsRNA genome since in their absence there was little synthesis of viral mRNA and dsRNA. The characterization of the kinetics of RNA transcription and replication of the viral genome under conditions where these proteins were silenced provided direct evidence for a second round of transcription during the replication of the virus. Interestingly, despite the decrease in mRNA accumulation when any of the four proteins was silenced, the synthesis of viral proteins decreased when VP2 and VP6 were knocked down, whereas the absence of VP1 and VP3 did not have a severe impact on viral protein synthesis. Characterization of viral particle assembly in the absence of VP1 and VP3 showed that while the formation of triple-layered particles and DLPs was decreased, the amount of assembled lower-density particles, often referred to as empty particles, was not different from the amount in control-infected cells, suggesting that viral particles can assemble in the absence of either VP1 or VP3. PMID- 19553304 TI - Vaccinia virus H7 protein contributes to the formation of crescent membrane precursors of immature virions. AB - Crescent membranes are the first viral structures that can be discerned during poxvirus morphogenesis. The crescents consist of a lipoprotein membrane and an outer lattice scaffold, which provides uniform curvature. Relatively little is known regarding the composition of the crescent membrane or its mode of formation. Here, we show that the H7 protein, which is conserved in all vertebrate poxviruses but has no discernible functional motifs or nonpoxvirus homologs, contributes to the formation of crescents and immature virions. Synthesis of the 17-kDa H7 protein was dependent on DNA replication and occurred late during vaccinia virus infection. Unlike many late proteins, however, H7 was not incorporated into mature virions or localized in cellular organelles. To gain insight into the role of H7, an inducible mutant was constructed and shown to have a conditional lethal phenotype: H7 expression and infectious virus formation were dependent on isopropyl-beta-D-thiogalactopyranoside. In the absence of inducer, viral late proteins were made, but membrane and core proteins were not processed by the I7 protease. A block in morphogenesis was demonstrated by transmission electron microscopy: neither typical crescents nor immature virions were detected in the absence of inducer. Instead, factory areas of the cytoplasm contained large, electron-dense inclusions, some of which had partially coated membrane segments at their surfaces. Separate, lower-density inclusions containing the D13 scaffold protein and endoplasmic reticulum membranes were also present. These features are most similar to those previously seen when expression of A11, another conserved nonvirion protein, is repressed. PMID- 19553305 TI - Inhibition of intrahepatic gamma interferon production by hepatitis C virus nonstructural protein 5A in transgenic mice. AB - Hepatitis C virus (HCV) utilizes strategies to suppress or evade the host immune response for establishment of persistent infection. We have shown previously that HCV nonstructural protein 5A (NS5A) impairs tumor necrosis factor alpha (TNF alpha)-mediated apoptosis. In this study, we have examined the immunomodulatory role of HCV NS5A protein in transgenic mouse (NS5A-Tg) liver when mice were challenged with an unrelated hepatotropic adenovirus as a nonspecific stimulus. Hepatotropic adenovirus was introduced intravenously into NS5A-Tg mice and control mice, and virus clearance from liver was compared over a time course of 3 weeks. The differential mRNA expression levels of 84 cytokine-related genes, signal pathway molecules, transcription factors, and cell surface molecules were determined using real-time reverse transcription-PCR array. NS5A-Tg mice failed to clear adenovirus from liver up to 3 weeks postinfection while control mice cleared virus within 1 to 2 weeks. Subsequent study revealed that gamma interferon (IFN-gamma) expression is inhibited at both the mRNA and protein levels in NS5A-Tg mice, and an inverse expression of transcription factors Gata-3 and Tbx21 is observed. However, TNF-alpha mRNA and protein expression were elevated in both NS5A-Tg and control mice. Together, our results suggested that HCV NS5A acts as an immunomodulator by inhibiting IFN-gamma production and may play an important role toward establishment of chronic HCV infection. PMID- 19553306 TI - T-cell tolerance for variability in an HLA class I-presented influenza A virus epitope. AB - To escape immune recognition, viruses acquire amino acid substitutions in class I human leukocyte antigen (HLA)-presented cytotoxic T-lymphocyte (CTL) epitopes. Such viral escape mutations may (i) prevent peptide processing, (ii) diminish class I HLA binding, or (iii) alter T-cell recognition. Because residues 418 to 426 of the hypervariable influenza A virus nucleoprotein (NP(418-426)) epitope are consistently bound by class I HLA and presented to CTL, we assessed the impact that intraepitope sequence variability has upon T-cell recognition. CTL elicited by intranasal influenza virus infection were tested for their cross recognition of 20 natural NP(418-426) epitope variants. Six of the variant epitopes, of both H1N1 and H3N2 origin, were cross-recognized by CTL while the remaining NP(418-426) epitope variants escaped targeting. A pattern emerged whereby variability at position 5 (P5) within the epitope reduced T-cell recognition, changes at P4 or P6 enabled CTL escape, and a mutation at P8 enhanced T-cell recognition. These data demonstrate that substitutions at P4 and/or P6 facilitate influenza virus escape from T-cell recognition and provide a model for the number, nature, and location of viral mutations that influence T cell cross-recognition. PMID- 19553308 TI - The efficacy of antigen processing is critical for protection against cytomegalovirus disease in the presence of viral immune evasion proteins. AB - Cytomegaloviruses (CMVs) code for immunoevasins, glycoproteins that are specifically dedicated to interfere with the presentation of antigenic peptides to CD8 T cells. Nonetheless, the biological outcome is not an immune evasion of the virus, since CD8 T cells can control CMV infection even when immunoevasins are expressed. Here, we compare the processing of a protective and a nonprotective epitope derived from the same viral protein, the antiapoptotic protein M45 in the murine model. The data provide evidence to conclude that protection against CMVs critically depends on antigenic peptides generated in an amount sufficient to exhaust the inhibitory capacity of immunoevasins. PMID- 19553307 TI - Protective efficacy of a single immunization of a chimeric adenovirus vector based vaccine against simian immunodeficiency virus challenge in rhesus monkeys. AB - Rare serotype and chimeric recombinant adenovirus (rAd) vectors that evade anti Ad5 immunity are currently being evaluated as potential vaccine vectors for human immunodeficiency virus type 1 and other pathogens. We have recently reported that a heterologous rAd prime-boost regimen expressing simian immunodeficiency virus (SIV) Gag afforded durable partial immune control of an SIV challenge in rhesus monkeys. However, single-shot immunization may ultimately be preferable for global vaccine delivery. We therefore evaluated the immunogenicity and protective efficacy of a single immunization of chimeric rAd5 hexon hypervariable region 48 (rAd5HVR48) vectors expressing SIV Gag, Pol, Nef, and Env against a homologous SIV challenge in rhesus monkeys. Inclusion of Env resulted in improved control of peak and set point SIV RNA levels following challenge. In contrast, DNA vaccine priming did not further improve the protective efficacy of rAd5HVR48 vectors in this system. PMID- 19553309 TI - Computational analysis of human adenovirus type 22 provides evidence for recombination among species D human adenoviruses in the penton base gene. AB - Recombination in human adenoviruses (HAdV) may confer virulence upon an otherwise nonvirulent strain. The genome sequence of species D HAdV type 22 (HAdV-D22) revealed evidence for recombination with HAdV-D19 and HAdV-D37 within the capsid penton base gene. Bootscan analysis demonstrated that recombination sites within the penton base gene frame the coding sequences for the two external hypervariable loops in the protein. A similar pattern of recombination was evident within other HAdV-D types but not other HAdV species. Further study of recombination among HAdVs is needed to better predict possible recombination events among wild-type viruses and adenoviral gene therapy vectors. PMID- 19553310 TI - The ubiquitin-specific peptidase USP15 regulates human papillomavirus type 16 E6 protein stability. AB - Proteomic identification of human papillomavirus type 16 (HPV16) E6-interacting proteins revealed several proteins involved in ubiquitin-mediated proteolysis. In addition to the well-characterized E6AP ubiquitin-protein ligase, a second HECT domain protein (HERC2) and a deubiquitylating enzyme (USP15) were identified by tandem affinity purification of HPV16 E6-associated proteins. This study focuses on the functional consequences of the interaction of E6 with USP15. Overexpression of USP15 resulted in increased levels of the E6 protein, and the small interfering RNA-mediated knockdown of USP15 decreased E6 protein levels. These results implicate USP15 directly in the regulation of E6 protein stability and suggest that ubiquitylated E6 could be a substrate for USP15 ubiquitin peptidase activity. It remains possible that E6 could affect the activity of USP15 on specific cellular substrates, a hypothesis that can be tested as more is learned about the substrates and pathways controlled by USP15. PMID- 19553311 TI - A structure-guided mutational analysis of simian virus 40 large T antigen: identification of surface residues required for viral replication and transformation. AB - Simian virus 40 large T antigen (TAg) transforms cells in culture and induces tumors in rodents. Genetic studies suggest that TAg interaction with the chaperone hsp70 and tumor suppressors pRb and p53 may not be sufficient to elicit complete transformation of cells. In order to identify additional cellular factors important for transformation, we designed mutations on the solvent exposed surface of TAg. We hypothesized that surface residues would interact directly with cellular targets and that the mutation of these residues might disrupt this interaction without perturbing TAg's global structure. Using structural data, we identified 61 amino acids on the surface of TAg. Each surface amino acid was changed to an alanine. Furthermore, five patches containing clusters of charged amino acids on the surface of TAg were identified. Within these patches, we selectively mutated three to four charged amino acids and thus generated five mutants (patch mutants 1 to 5). We observed that while patch mutants 3 and 4 induced foci in REF52 cells, patch mutants 1 and 2 were deficient in focus formation. We determined that the patch 1 mutant is defective in p53 binding, thus explaining its defect in transformation. The patch 2 mutant can interact with the Rb family members and p53 like wild-type TAg but is unable to transform cells, suggesting that it is defective for action on an unknown cellular target essential for transformation. Our results suggest that the histone acetyltransferase CBP/p300 is one of the potential targets affected by the mutations in patch 2. PMID- 19553312 TI - Palmitoylation of the influenza A virus M2 protein is not required for virus replication in vitro but contributes to virus virulence. AB - The influenza A virus M2 protein has important roles during virus entry and in the assembly of infectious virus particles. The cytoplasmic tail of the protein can be palmitoylated at a cysteine residue, but this residue is not conserved in a number of human influenza A virus isolates. Recombinant viruses encoding M2 proteins with a serine substituted for the cysteine at position 50 were generated in the A/WSN/33 (H1N1) and A/Udorn/72 (H3N2) genetic backgrounds. The recombinant viruses were not attenuated for replication in MDCK cells, Calu-3 cells, or in primary differentiated murine trachea epithelial cell cultures, indicating there was no significant contribution of M2 palmitoylation to virus replication in vitro. The A/WSN/33 M2C50S virus displayed a slightly reduced virulence after infection of mice, suggesting that there may be novel functions for M2 palmitoylation during in vivo infection. PMID- 19553313 TI - Mixed infection and the genesis of influenza virus diversity. AB - The emergence of viral infections with potentially devastating consequences for human health is highly dependent on their underlying evolutionary dynamics. One likely scenario for an avian influenza virus, such as A/H5N1, to evolve to one capable of human-to-human transmission is through the acquisition of genetic material from the A/H1N1 or A/H3N2 subtypes already circulating in human populations. This would require that viruses of both subtypes coinfect the same cells, generating a mixed infection, and then reassort. Determining the nature and frequency of mixed infection with influenza virus is therefore central to understanding the emergence of pandemic, antigenic, and drug-resistant strains. To better understand the potential for such events, we explored patterns of intrahost genetic diversity in recently circulating strains of human influenza virus. By analyzing multiple viral genome sequences sampled from individual influenza patients we reveal a high level of mixed infection, including diverse lineages of the same influenza virus subtype, drug-resistant and -sensitive strains, those that are likely to differ in antigenicity, and even viruses of different influenza virus types (A and B). These results reveal that individuals can harbor influenza viruses that differ in major phenotypic properties, including those that are antigenically distinct and those that differ in their sensitivity to antiviral agents. PMID- 19553314 TI - Proteolytic activation of the spike protein at a novel RRRR/S motif is implicated in furin-dependent entry, syncytium formation, and infectivity of coronavirus infectious bronchitis virus in cultured cells. AB - The spike (S) protein of the coronavirus (CoV) infectious bronchitis virus (IBV) is cleaved into S1 and S2 subunits at the furin consensus motif RRFRR(537)/S in virus-infected cells. In this study, we observe that the S2 subunit of the IBV Beaudette strain is additionally cleaved at the second furin site (RRRR(690)/S) in cells expressing S constructs and in virus-infected cells. Detailed time course experiments showed that a peptide furin inhibitor, decanoyl-Arg-Val-Lys Arg-chloromethylketone, blocked both viral entry and syncytium formation. Site directed mutagenesis studies revealed that the S1/S2 cleavage by furin was not necessary for, but could promote, syncytium formation by and infectivity of IBV in Vero cells. In contrast, the second site is involved in the furin dependence of viral entry and syncytium formation. Mutations of the second site from furin cleavable RRRR/S to non-furin-cleavable PRRRS and AAARS, respectively, abrogated the furin dependence of IBV entry. Instead, a yet-to-be-identified serine protease(s) was involved, as revealed by protease inhibitor studies. Furthermore, sequence analysis of CoV S proteins by multiple alignments showed conservation of an XXXR/S motif, cleavable by either furin or other trypsin-like proteases, at a position equivalent to the second IBV furin site. Taken together, these results suggest that proteolysis at a novel XXXR/S motif in the S2 subunit might be a common mechanism for the entry of CoV into cells. PMID- 19553315 TI - Is single-strand conformation polymorphism analysis of the full 5' untranslated region an adequate approach to study hepatitis C virus quasispecies distribution? AB - Single-strand conformation polymorphism (SSCP) analysis is used by many laboratories to study the quasispecies distribution of the hepatitis C virus (HCV). Here we question the validity of this experimental approach, as conclusions are drawn from the analysis of the migration patterns of two ssDNA molecules and not from RNA. Using previously characterized mutants of the HCV 5' untranslated regions, we show that contrary to what has been predicted, SSCP migration patterns of DNA amplicons with differences in their nucleotide sequences generated from the full 5' UTR of HCV are not necessarily unique. PMID- 19553316 TI - Previously unrecognized amino acid substitutions in the hemagglutinin and fusion proteins of measles virus modulate cell-cell fusion, hemadsorption, virus growth, and penetration rate. AB - Wild-type measles virus (MV) isolated in B95a cells could be adapted to Vero cells after several blind passages. In this study, we have determined the complete nucleotide sequences of the genomes of the wild type (T11wild) and its Vero cell-adapted (T11Ve-23) MV strain and identified amino acid substitutions R516G, E271K, D439E and G464W (D439E/G464W), N481Y/H495R, and Y187H/L204F in the nucleocapsid, V, fusion (F), hemagglutinin (H), and large proteins, respectively. Expression of mutated H and F proteins from cDNA revealed that the H495R substitution, in addition to N481Y, in the H protein was necessary for the wild type H protein to use CD46 efficiently as a receptor and that the G464W substitution in the F protein was important for enhanced cell-cell fusion. Recombinant wild-type MV strains harboring the F protein with the mutations D439E/G464W [F(D439E/G464W)] and/or H(N481Y/H495R) protein revealed that both mutated F and H proteins were required for efficient syncytium formation and virus growth in Vero cells. Interestingly, a recombinant wild-type MV strain harboring the H(N481Y/H495R) protein penetrated slowly into Vero cells, while a recombinant wild-type MV strain harboring both the F(D439E/G464W) and H(N481Y/H495R) proteins penetrated efficiently into Vero cells, indicating that the F(D439E/G464W) protein compensates for the inefficient penetration of a wild type MV strain harboring the H(N481Y/H495R) protein. Thus, the F and H proteins synergistically function to ensure efficient wild-type MV growth in Vero cells. PMID- 19553317 TI - Brd4 regulation of papillomavirus protein E2 stability. AB - The papillomavirus (PV) E2 protein is an important regulator of the viral life cycle. It has diverse roles in viral transcription, DNA replication, and genome maintenance. Our laboratory has previously identified the cellular bromodomain protein Brd4 as a key interacting partner of E2. Brd4 mediates the transcriptional activation function of E2 and plays an important role in viral genome maintenance in dividing cells. E2 interacts with the C-terminal domain (CTD) of Brd4, and the CTD functions in a dominant-negative manner through binding E2 and interfering with E2's interaction with the full-length Brd4 protein. Previous studies have shown that PV E2 proteins are short lived; however, the mechanisms regulating their stability and degradation have not yet been well established. In this study, we explored the role of Brd4 in the regulation of bovine PV 1 (BPV1) and human PV 16 (HPV16) E2 stability. Expression of the Brd4 CTD dramatically increases E2 levels. Both BPV1 E2 and HPV16 E2 are regulated by ubiquitylation, and Brd4 CTD expression blocks this ubiquitylation, thus stabilizing the E2 protein. Furthermore, we have identified the cullin-based E3 ligases and specifically cullin-3 as potential components of the ubiquitylation machinery that targets both BPV1 and HPV16 E2 for ubiquitylation. Expression of the Brd4 CTD blocks the interaction between E2 and the cullin-3 complex. In addition to Brd4's role in mediating E2 transcription and genome tethering activities, these data suggest a potential role for Brd4 in regulating E2 stability and protein levels within PV-infected cells. PMID- 19553318 TI - Subtype-specific differences in the human immunodeficiency virus type 1 reverse transcriptase connection subdomain of CRF01_AE are associated with higher levels of resistance to 3'-azido-3'-deoxythymidine. AB - We previously shown that mutations in the connection (CN) subdomain of human immunodeficiency virus type 1 (HIV-1) subtype B reverse transcriptase (RT) increase 3'-azido-3'-deoxythymidine (AZT) resistance in the context of thymidine analog mutations (TAMs) by affecting the balance between polymerization and RNase H activity. To determine whether this balance affects drug resistance in other HIV-1 subtypes, recombinant subtype CRF01_AE was analyzed. Interestingly, CRF01_AE containing TAMs exhibited 64-fold higher AZT resistance relative to wild type B, whereas AZT resistance of subtype B containing the same TAMs was 13-fold higher, which in turn correlated with higher levels of AZT-monophosphate (AZTMP) excision on both RNA and DNA templates. The high level of AZT resistance exhibited by CRF01_AE was primarily associated with the T400 residue in wild-type subtype AE CN subdomain. An A400T substitution in subtype B enhanced AZT resistance, increased AZTMP excision on both RNA and DNA templates, and reduced RNase H cleavage. Replacing the T400 residue in CRF01_AE with alanine restored AZT sensitivity and reduced AZTMP excision on both RNA and DNA templates, suggesting that the T400 residue increases AZT resistance in CRF01_AE at least in part by directly increasing the efficiency of AZTMP excision. These results show for the first time that CRF01_AE exhibits higher levels of AZT resistance in the presence of TAMs and that this resistance is primarily associated with T400. Our results also show that mixing the RT polymerase, CN, and RNase H domains from different subtypes can underestimate AZT resistance levels, and they emphasize the need to develop subtype-specific genotypic and phenotypic assays to provide more accurate estimates of clinical drug resistance. PMID- 19553319 TI - Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 K-bZIP modulates latency-associated nuclear protein-mediated suppression of lytic origin-dependent DNA synthesis. AB - The original cotransfection replication assay identified eight human herpesvirus 8 (HHV8)-encoded proteins required for origin-dependent lytic DNA replication. Previously, we demonstrated that under conditions where K-Rta is overexpressed, a K-bZIP knockout bacmid displayed an aberrant subcellular localization pattern for the latency-associated nuclear protein (LANA). Additionally, these same studies demonstrated that K-bZIP interacts with LANA in the absence of K-Rta and that K bZIP does not directly participate in, but may facilitate, the initiation of lytic DNA synthesis. We developed a modification of the transient cotransfection replication assay wherein both lytic (oriLyt) and latent (terminal repeat) DNA replication are evaluated simultaneously. We now show that LANA represses origin dependent lytic DNA replication in a dose dependent manner when added to the cotransfection replication assay. This repression was overcome by increasing amounts of a K-bZIP expression plasmid in the cotransfection mixture or by dominant-negative inhibition of the interaction of LANA with K-bZIP by the overexpression of the K-bZIP-LANA binding domain. Chromatin immunoprecipitation assays show that LANA interacts with oriLyt in the absence of K-bZIP expression, suggesting that suppression of lytic replication by LANA is mediated by direct binding. The interaction of K-bZIP with oriLyt was dependent upon the expression of LANA; however, LANA interacted with oriLyt independently of K-bZIP expression. These data suggest that the interaction of LANA with K-bZIP modulates lytic and latent replication and that K-bZIP facilitates lytic DNA replication and modulates the switch from the latent phase of the virus. PMID- 19553321 TI - Characterization of the H5N1 highly pathogenic avian influenza virus derived from wild pikas in China. AB - The highly pathogenic H5N1 avian influenza virus emerged from China in 1996 and has spread across Eurasia and Africa, with a continuous stream of new cases of human infection appearing since the first large-scale outbreak among migratory birds at Qinghai Lake. The role of wild birds, which are the natural reservoirs for the virus, in the epidemiology of the H5N1 virus has raised great public health concern, but their role in the spread of the virus within the natural ecosystem of free-ranging terrestrial wild mammals remains unclear. In this study, we investigated H5N1 virus infection in wild pikas in an attempt to trace the circulation of the virus. Seroepidemiological surveys confirmed a natural H5N1 virus infection of wild pikas in their native environment. The hemagglutination gene of the H5N1 virus isolated from pikas reveals two distinct evolutionary clades, a mixed/Vietnam H5N1 virus sublineage (MV-like pika virus) and a wild bird Qinghai (QH)-like H5N1 virus sublineage (QH-like pika virus). The amino acid residue (glutamic acid) at position 627 encoded by the PB2 gene of the MV-like pika virus was different from that of the QH-like pika virus; the residue of the MV-like pika virus was the same as that of the goose H5N1 virus (A/GS/Guangdong [GD]/1/96). Further, we discovered that in contrast to the MV like pika virus, which is nonpathogenic to mice, the QH-like pika virus is highly pathogenic. To mimic the virus infection of pikas, we intranasally inoculated rabbits, a species closely related to pikas, with the H5N1 virus of pika origin. Our findings first demonstrate that wild pikas are mammalian hosts exposed to H5N1 subtype avian influenza viruses in the natural ecosystem and also imply a potential transmission of highly pathogenic avian influenza virus from wild mammals into domestic mammalian hosts and humans. PMID- 19553320 TI - Impact of quaternary organization on the antigenic structure of the tick-borne encephalitis virus envelope glycoprotein E. AB - The envelope protein E of flaviviruses mediates both receptor-binding and membrane fusion. At the virion surface, 180 copies of E are tightly packed and organized in a herringbone-like icosahedral structure, whereas in noninfectious subviral particles, 60 copies are arranged in a T=1 icosahedral symmetry. In both cases, the basic building block is an E dimer which exposes the binding sites for neutralizing antibodies at its surface. It was the objective of our study to assess the dependence of the antigenic structure of E on its quaternary arrangement, i.e., as part of virions, recombinant subviral particles, or soluble dimers. For this purpose, we used a panel of 11 E protein-specific neutralizing monoclonal antibodies, mapped to distinct epitopes in each of the three E protein domains, and studied their reactivity with the different soluble and particulate forms of tick-borne encephalitis virus E protein under nondenaturing immunoassay conditions. Significant differences in the reactivities with these forms were observed that could be related to (i) limited access of certain epitopes at the virion surface; (ii) limited occupancy of epitopes in virions due to steric hindrance between antibodies; (iii) differences in the avidity to soluble forms compared to the virion, presumably related to the flexibility of E at its domain junctions; and (iv) modulations of the external E protein surface through interactions with its stem-anchor structure. We have thus identified several important factors that influence the antigenicity of the flavivirus E protein and have an impact on the interaction with neutralizing antibodies. PMID- 19553322 TI - The CREB site in the proximal enhancer is critical for cooperative interaction with the other transcription factor binding sites to enhance transcription of the major intermediate-early genes in human cytomegalovirus-infected cells. AB - One of the two SP1 sites in the proximal enhancer of the human cytomegalovirus (HCMV) major immediate-early (MIE) promoter is essential for transcription in human fibroblast cells (H. Isomura, M. F. Stinski, A. Kudoh, T. Daikoku, N. Shirata, and T. Tsurumi, J. Virol. 79:9597-9607, 2005). Upstream of the two SP1 sites to -223 relative to the +1 transcription start site, there are an additional five DNA binding sites for eukaryotic transcription factors. We determined the effects of the various transcription factor DNA binding sites on viral MIE RNA transcription, viral gene expression, viral DNA synthesis, or infectious virus production. We prepared recombinant HCMV bacterial artificial chromosome (BAC) DNAs with either one site missing or one site present upstream of the two SP1 sites. Infectious recombinant HCMV BAC DNAs were transfected into various cell types to avoid the effect of the virion-associated transactivators. Regardless of the cell type, which included human fibroblast, endothelial, and epithelial cells, the CREB site had the most significant and independent effect on the MIE promoter. The other sites had a minor independent effect. However, the combination of the different transcription factor DNA binding sites was significantly stronger than multiple duplications of the CREB site. These findings indicate that the CREB site in the presence of the other sites has a major role for the replication of HCMV. PMID- 19553324 TI - Mutational analysis of the herpes simplex virus type 1 DNA packaging protein UL33. AB - The UL33 protein of herpes simplex virus type 1 (HSV-1) is thought to be a component of the terminase complex that mediates the cleavage and packaging of viral DNA. In this study we describe the generation and characterization of a series of 15 UL33 mutants containing insertions of five amino acids located randomly throughout the 130-residue protein. Of these mutants, seven were unable to complement the growth of the UL33-null virus dlUL33 in transient assays and also failed to support the cleavage and packaging of replicated amplicon DNA into capsids. The insertions in these mutants were clustered between residues 51 and 74 and between 104 and 116, within the most highly conserved regions of the protein. The ability of the mutants to interact with the UL28 component of the terminase was assessed in immunoprecipitation and immunofluorescence assays. All four mutants with insertions between amino acids 51 and 74 were impaired in this interaction, whereas two of the three mutants in the second region (with insertions at positions 111 and 116) were not affected. These data indicate that the ability of UL33 to interact with UL28 is probably necessary, but not sufficient, to support viral growth and DNA packaging. PMID- 19553323 TI - Virus entry via the alternative coreceptors CCR3 and FPRL1 differs by human immunodeficiency virus type 1 subtype. AB - Human immunodeficiency virus type 1 (HIV-1) infects target cells by binding to CD4 and a chemokine receptor, most commonly CCR5. CXCR4 is a frequent alternative coreceptor (CoR) in subtype B and D HIV-1 infection, but the importance of many other alternative CoRs remains elusive. We have analyzed HIV-1 envelope (Env) proteins from 66 individuals infected with the major subtypes of HIV-1 to determine if virus entry into highly permissive NP-2 cell lines expressing most known alternative CoRs differed by HIV-1 subtype. We also performed linear regression analysis to determine if virus entry via the major CoR CCR5 correlated with use of any alternative CoR and if this correlation differed by subtype. Virus pseudotyped with subtype B Env showed robust entry via CCR3 that was highly correlated with CCR5 entry efficiency. By contrast, viruses pseudotyped with subtype A and C Env proteins were able to use the recently described alternative CoR FPRL1 more efficiently than CCR3, and use of FPRL1 was correlated with CCR5 entry. Subtype D Env was unable to use either CCR3 or FPRL1 efficiently, a unique pattern of alternative CoR use. These results suggest that each subtype of circulating HIV-1 may be subject to somewhat different selective pressures for Env-mediated entry into target cells and suggest that CCR3 may be used as a surrogate CoR by subtype B while FPRL1 may be used as a surrogate CoR by subtypes A and C. These data may provide insight into development of resistance to CCR5 targeted entry inhibitors and alternative entry pathways for each HIV-1 subtype. PMID- 19553326 TI - Therapeutic memory T cells require costimulation for effective clearance of a persistent viral infection. AB - Persistent viral infections are a major health concern worldwide. During persistent infection, overwhelming viral replication and the rapid loss of antiviral T-cell function can prevent immune-mediated clearance of the infection, and therapies to reanimate the immune response and purge persistent viruses have been largely unsuccessful. Adoptive immunotherapy using memory T cells is a highly successful therapeutic approach to eradicate a persistent viral infection. Understanding precisely how therapeutically administered memory T cells achieve clearance should improve our ability to terminate states of viral persistence in humans. Mice persistently infected from birth with lymphocytic choriomeningitis virus are tolerant to the pathogen at the T-cell level and thus provide an excellent model to evaluate immunotherapeutic regimens. Previously, we demonstrated that adoptively transferred memory T cells require recipient dendritic cells to effectively purge an established persistent viral infection. However, the mechanisms that reactivate and sustain memory T-cell responses during clearance of such an infection remain unclear. Here we establish that therapeutic memory T cells require CD80 and CD86 costimulatory signals to efficiently clear an established persistent viral infection in vivo. Early blockade of costimulatory pathways with CTLA-4-Fc decreased the secondary expansion of virus-specific CD8(+) and CD4(+) memory T cells as well as their ability to produce antiviral cytokines and purge the persistent infection. Late costimulation blockade also reduced virus-specific T-cell numbers, illustrating that sustained interactions with costimulatory molecules is required for efficient T-cell expansion. These findings indicate that antiviral memory T cells require costimulation to efficiently clear a persistent viral infection and that costimulatory pathways can be targeted to modulate the magnitude of an adoptive immunotherapeutic regimen. PMID- 19553325 TI - Measurement of human immunodeficiency virus type 1 preintegration transcription by using Rev-dependent Rev-CEM cells reveals a sizable transcribing DNA population comparable to that from proviral templates. AB - Preintegration transcription is an early process in human immunodeficiency virus type 1 infection and has been suggested to occur at a low level. The templates have also been suggested to represent a small population of nonintegrated viral DNA, particularly the two-long-terminal-repeat (2-LTR) circles. However, these determinations were made by either using PCR amplification of viral transcripts in bulk cell populations or utilizing the LTR-driving reporter cells that measure the synthesis of Tat. The intrinsic leakiness of LTR often makes the measurement of low-level viral transcription inaccurate. Since preintegration transcription also generates Rev, to eliminate the nonspecificity associated with the use of LTR alone we have developed a novel Rev-dependent indicator cell, Rev-CEM, to measure preintegration transcription based on the amount of Rev generated. In this report, using Rev-CEM cells, we demonstrate that preintegration transcription occurs on a much larger scale than expected. The transcribing population derived from nonintegrated viral DNA was comparable (at approximately 70%) to that derived from provirus in a productive viral replication cycle. Nevertheless, each nonintegrated viral DNA template exhibited a significant reduction in the level of transcriptional activity in the absence of integration. We also performed flow cytometry sorting of infected cells to identify viral templates. Surprisingly, our results suggest that the majority of 2-LTR circles are not active in directing transcription. It is likely that the nonintegrated templates are from the predominant DNA species, such as the full-length, linear DNA. Our results also suggest that a nonintegrating lentiviral vector can be as effective as an integrating vector in directing gene expression in nondividing cells, with the proper choice of an internal promoter. PMID- 19553327 TI - Human immunodeficiency virus type 1 escapes from interleukin-2-producing CD4+ T cell responses without high-frequency fixation of mutations. AB - The presence of interleukin-2 (IL-2)-producing human immunodeficiency virus type 1 (HIV-1)-specific CD4(+) T-cell responses has been associated with the immunological control of HIV-1 replication; however, the causal relationship between these factors remains unclear. Here we show that IL-2-producing HIV-1 specific CD4(+) T cells can be cloned from acutely HIV-1-infected individuals. Despite the early presence of these cells, each of the individuals in the present study exhibited progressive disease, with one individual showing rapid progression. In this rapid progressor, three IL-2-producing HIV-1 Gag-specific CD4(+) T-cell responses were identified and mapped to the following optimal epitopes: HIVWASRELER, REPRGSDIAGT, and FRDYVDRFYKT. Responses to these epitopes in peripheral blood mononuclear cells were monitored longitudinally to >1 year postinfection, and contemporaneous circulating plasma viruses were sequenced. A variant of the FRDYVDRFYKT epitope sequence, FRDYVDQFYKT, was observed in 1/21 plasma viruses sequenced at 5 months postinfection and 1/10 viruses at 7 months postinfection. This variant failed to stimulate the corresponding CD4(+) T-cell clone and thus constitutes an escape mutant. Responses to each of the three Gag epitopes were rapidly lost, and this loss was accompanied by a loss of antigen specific cells in the periphery as measured by using an FRDYVDRFYKT-presenting major histocompatibility complex class II tetramer. Highly active antiretroviral therapy was associated with the reemergence of FRDYVDRFYKT-specific cells by tetramer. Thus, our data support that IL-2-producing HIV-1-specific CD4(+) T-cell responses can exert immune pressure during early HIV-1 infection but that the inability of these responses to enforce enduring control of viral replication is related to the deletion and/or dysfunction of HIV-1-specific CD4(+) T cells rather than to the fixation of escape mutations at high frequencies. PMID- 19553328 TI - A novel genotype H9N2 influenza virus possessing human H5N1 internal genomes has been circulating in poultry in eastern China since 1998. AB - Many novel reassortant influenza viruses of the H9N2 genotype have emerged in aquatic birds in southern China since their initial isolation in this region in 1994. However, the genesis and evolution of H9N2 viruses in poultry in eastern China have not been investigated systematically. In the current study, H9N2 influenza viruses isolated from poultry in eastern China during the past 10 years were characterized genetically and antigenically. Phylogenetic analysis revealed that these H9N2 viruses have undergone extensive reassortment to generate multiple novel genotypes, including four genotypes (J, F, K, and L) that have never been recognized before. The major H9N2 influenza viruses represented by A/Chicken/Beijing/1/1994 (Ck/BJ/1/94)-like viruses circulating in poultry in eastern China before 1998 have been gradually replaced by A/Chicken/Shanghai/F/1998 (Ck/SH/F/98)-like viruses, which have a genotype different from that of viruses isolated in southern China. The similarity of the internal genes of these H9N2 viruses to those of the H5N1 influenza viruses isolated from 2001 onwards suggests that the Ck/SH/F/98-like virus may have been the donor of internal genes of human and poultry H5N1 influenza viruses circulating in Eurasia. Experimental studies showed that some of these H9N2 viruses could be efficiently transmitted by the respiratory tract in chicken flocks. Our study provides new insight into the genesis and evolution of H9N2 influenza viruses and supports the notion that some of these viruses may have been the donors of internal genes found in H5N1 viruses. PMID- 19553329 TI - Bluetongue virus VP6 acts early in the replication cycle and can form the basis of chimeric virus formation. AB - A minor core protein, VP6, of bluetongue virus (BTV) possesses nucleoside triphosphatase, RNA binding, and helicase activities. Although the enzymatic functions of VP6 have been documented in vitro using purified protein, its definitive role in BTV replication remains unclear. In this study, using a recently developed T7 transcript-based reverse genetics system for BTV, we examined the importance of VP6 in virus replication. We show that VP6 is active early in replication, consistent with a role as part of the transcriptase or packaging complex, and that its action can be provided in trans by a newly developed complementary cell line. Furthermore, the genomic segment encoding VP6 was mutated to reveal the cis-acting sequences required for replication or packaging, which subsequently enabled the construction of a chimeric BTV expressing enhanced green fluorescent protein. These data confirm that one of the 10 genome segments of BTV can be replaced with a chimeric RNA containing the essential packaging and replication signals of BTV and the coding sequence of a foreign gene. PMID- 19553330 TI - Dexamethasone treatment of calves latently infected with bovine herpesvirus 1 leads to activation of the bICP0 early promoter, in part by the cellular transcription factor C/EBP-alpha. AB - Sensory neurons within trigeminal ganglia (TG) are the primary site for bovine herpesvirus 1 (BHV-1) latency. During latency, viral gene expression is restricted to the latency-related (LR) gene and the open reading frame ORF-E. We previously constructed an LR mutant virus that expresses LR RNA but not any of the known LR proteins. In contrast to calves latently infected with wild-type (wt) BHV-1 or the LR rescued virus, the LR mutant virus does not reactivate from latency following dexamethasone (DEX) treatment. In this study, we demonstrated that bICP0, but not bICP4, transcripts were consistently detected in TG of calves infected with the LR mutant or LR rescued virus following DEX treatment. Calves latently infected with the LR rescued virus but not the LR mutant virus expressed late transcripts, which correlated with shedding of infectious virus following DEX treatment. The bICP4 and bICP0 genes share a common immediate-early promoter, suggesting that this promoter was not consistently activated during DEX-induced reactivation from latency. The bICP0 gene also contains a novel early promoter that was activated by DEX in mouse neuroblastoma cells. Expression of a cellular transcription factor, C/EBP-alpha, was stimulated by DEX, and C/EBP-alpha expression was necessary for DEX induction of bICP0 early promoter activity. C/EBP-alpha directly interacted with bICP0 early promoter sequences that were necessary for trans activation by C/EBP-alpha. In summary, DEX treatment of latently infected calves induced cellular factors that stimulated bICP0 early promoter activity. Activation of bICP0 early promoter activity does not necessarily lead to late gene expression and virus shedding. PMID- 19553331 TI - gp340 promotes transcytosis of human immunodeficiency virus type 1 in genital tract-derived cell lines and primary endocervical tissue. AB - The human scavenger receptor gp340 has been identified as a binding protein for the human immunodeficiency virus type 1 (HIV-1) envelope that is expressed on the cell surface of female genital tract epithelial cells. This interaction allows such epithelial cells to efficiently transmit infective virus to susceptible targets and maintain viral infectivity for several days. Within the context of vaginal transmission, HIV must first traverse a normally protective mucosa containing a cell barrier to reach the underlying T cells and dendritic cells, which propagate and spread the infection. The mechanism by which HIV-1 can bypass an otherwise healthy cellular barrier remains an important area of study. Here, we demonstrate that genital tract-derived cell lines and primary human endocervical tissue can support direct transcytosis of cell-free virus from the apical to basolateral surfaces. Further, this transport of virus can be blocked through the addition of antibodies or peptides that directly block the interaction of gp340 with the HIV-1 envelope, if added prior to viral pulsing on the apical side of the cell or tissue barrier. Our data support a role for the previously described heparan sulfate moieties in mediating this transcytosis but add gp340 as an important facilitator of HIV-1 transcytosis across genital tract tissue. This study demonstrates that HIV-1 actively traverses the protective barriers of the human genital tract and presents a second mechanism whereby gp340 can promote heterosexual transmission. PMID- 19553332 TI - A New World primate deficient in tetherin-mediated restriction of human immunodeficiency virus type 1. AB - Human immunodeficiency virus type 1 (HIV-1) does not replicate in primary cells of New World primates. To better understand this restriction, we expressed owl monkey (Aotus nancymaae) CD4 and CXCR4 in the owl monkey kidney cell line, OMK. An HIV-1 variant modified to evade the owl monkey restriction factor TRIM-cyp replicated efficiently in these cells but could not replicate in primary A. nancymaae CD4-positive T cells. To understand this difference, we examined APOBEC3G and tetherin orthologs from OMK cells and primary A. nancymaae cells. We observed that OMK cells expressed substantially lower levels of APOBEC3G than did A. nancymaae cells. A. nancymaae, but not marmoset (Callithrix jacchus), APOBEC3G was partially downregulated by HIV-1 vif and reduced but did not abolish HIV-1 replication when stably expressed in OMK cells. The functional difference between A. nancymaae and marmoset APOBEC3Gs mapped to residue 128, previously shown to distinguish African green monkey from human APOBEC3G. We also characterized tetherin orthologs from OMK and A. nancymaae cells. The A. nancymaae tetherin ortholog, but not OMK tetherin, prevented HIV-1 release. Alteration of threonine 181 of OMK tetherin rescued its function and its efficient N glycosylation. All alleles of Aotus lemurinus griseimembra examined, but none of A. nancymaae or Aotus vociferans, encoded this nonfunctional tetherin ortholog. Our data indicate that HIV-1 replication in owl monkeys is not restricted at entry but can be limited by APOBEC3G and tetherin. Further, A. lemurinus griseimembra does not restrict HIV-1 replication via tetherin, a property likely useful for the study of tetherin-restricted viruses. PMID- 19553333 TI - Site-specific phosphorylation regulates human T-cell leukemia virus type 2 Rex function in vivo. AB - Human T-cell leukemia virus type 2 (HTLV-2) Rex is a transacting regulatory protein required for efficient cytoplasmic expression of the unspliced and incompletely spliced viral mRNA transcripts encoding the structural and enzymatic proteins. Previously, it was demonstrated that phosphorylation of Rex-2, predominantly on serine residues, is correlated with an altered conformation, as observed by a gel mobility shift and the detection of two related protein species (p24(Rex) and p26(Rex)). Rex-2 phosphorylation is required for specific binding to its viral-mRNA target sequence and inhibition of mRNA splicing and may be linked to subcellular compartmentalization. Thus, the phosphorylation-induced structural state of Rex in the infected cell may be a switch that determines whether HTLV exists in a latent or productive state. We conducted a phosphoryl and functional mapping of both structural forms of mammalian-cell-expressed Rex 2 using affinity purification, liquid chromatography-tandem mass spectrometry, and site-directed substitutional mutational analysis. We identified two phosphorylation sites in p24(Rex) at Ser-117 and Thr-164. We also identified six phosphorylation sites in p26(Rex) at Thr-19, Ser-117, Ser-125, Ser-151, Ser-153, and Thr-164. We evaluated the functional significance of these phosphorylation events and found that phosphorylation on Thr-164, Ser-151, and Ser-153 is critical for Rex-2 function in vivo and that phosphorylation of Ser-151 is correlated with nuclear/nucleolar subcellular localization. Overall, this work is the first to completely map the phosphorylation sites in Rex-2 and provides important insight into the phosphorylation continuum that tightly regulates Rex-2 structure, cellular localization, and function. PMID- 19553334 TI - Differential rates of protein folding and cellular trafficking for the Hendra virus F and G proteins: implications for F-G complex formation. AB - Hendra virus F protein-promoted membrane fusion requires the presence of the viral attachment protein, G. However, events leading to the association of these glycoproteins remain unclear. Results presented here demonstrate that Hendra virus G undergoes slower secretory pathway trafficking than is observed for Hendra virus F. This slowed trafficking is not dependent on the G protein cytoplasmic tail, the presence of the G receptor ephrin B2, or interaction with other viral proteins. Instead, Hendra virus G was found to undergo intrinsically slow oligomerization within the endoplasmic reticulum. These results suggest that the critical F-G interactions occur only after the initial steps of synthesis and cellular transport. PMID- 19553335 TI - Antibody specificities associated with neutralization breadth in plasma from human immunodeficiency virus type 1 subtype C-infected blood donors. AB - Defining the specificities of the anti-human immunodeficiency virus type 1 (HIV 1) envelope antibodies able to mediate broad heterologous neutralization will assist in identifying targets for an HIV-1 vaccine. We screened 70 plasmas from chronically HIV-1-infected individuals for neutralization breadth. Of these, 16 (23%) were found to neutralize 80% or more of the viruses tested. Anti-CD4 binding site (CD4bs) antibodies were found in almost all plasmas independent of their neutralization breadth, but they mainly mediated neutralization of the laboratory strain HxB2 with little effect on the primary virus, Du151. Adsorption with Du151 monomeric gp120 reduced neutralizing activity to some extent in most plasma samples when tested against the matched virus, although these antibodies did not always confer cross-neutralization. For one plasma, this activity was mapped to a site overlapping the CD4-induced (CD4i) epitope and CD4bs. Anti membrane-proximal external region (MPER) (r = 0.69; P < 0.001) and anti-CD4i (r = 0.49; P < 0.001) antibody titers were found to be correlated with the neutralization breadth. These anti-MPER antibodies were not 4E10- or 2F5-like but spanned the 4E10 epitope. Furthermore, we found that anti-cardiolipin antibodies were correlated with the neutralization breadth (r = 0.67; P < 0.001) and anti MPER antibodies (r = 0.6; P < 0.001). Our study suggests that more than one epitope on the envelope glycoprotein is involved in the cross-reactive neutralization elicited during natural HIV-1 infection, many of which are yet to be determined, and that polyreactive antibodies are possibly involved in this phenomenon. PMID- 19553336 TI - Bluetongue virus targets conventional dendritic cells in skin lymph. AB - Bluetongue virus (BTV) is the etiological agent of bluetongue, a hemorrhagic disease of ruminants (particularly sheep), which causes important economic losses around the world. BTV is transmitted primarily via the bites of infected midges, which inject the virus into the ruminant's skin during blood feeding. The virus initially replicates in the draining lymph node and then disseminates to secondary organs where it induces edema, hemorrhages, and necrosis. In this study, we show that ovine conventional dendritic cells (cDCs) are the primary targets of BTV that contribute to the primary dissemination of BTV from the skin to draining lymph nodes. Lymph cDCs support BTV RNA and protein synthesis, as well as the production of infectious virus belonging to several different BTV serotypes, regardless of their level of attenuation. Afferent lymph cell subsets, other than cDCs, showed only marginal levels of BTV protein expression. BTV infection provoked a massive recruitment of cDCs to the sheep skin and afferent lymph, providing cellular targets for infection. Although BTV productively infects cDCs, no negative impact on their physiology was detected. Indeed, BTV infection and protein expression in cDCs enhanced their survival rate. Several serotypes of BTV stimulated the surface expression of the CD80 and CD86 costimulatory molecules on cDCs as well as the mRNA synthesis of cytokines involved in inflammation and immunity, i.e., interleukin-12 (IL-12), IL-1beta, and IL-6. BTV-infected cDCs stimulated antigen-specific CD4 and CD8 proliferation as well as gamma interferon production. BTV initially targets cDCs while preserving their functional properties, reflecting the optimal adaptation of the virus to its host cells for its first spread. PMID- 19553337 TI - Toll-like receptor 4 deficiency increases disease and mortality after mouse hepatitis virus type 1 infection of susceptible C3H mice. AB - Severe acute respiratory syndrome (SARS) is characterized by substantial acute pulmonary inflammation with a high mortality rate. Despite the identification of SARS coronavirus (SARS-CoV) as the etiologic agent of SARS, a thorough understanding of the underlying disease pathogenesis has been hampered by the lack of a suitable animal model that recapitulates the human disease. Intranasal (i.n.) infection of A/J mice with the CoV mouse hepatitis virus strain 1 (MHV-1) induces an acute respiratory disease with a high lethality rate that shares several pathological similarities with SARS-CoV infection in humans. In this study, we examined virus replication and the character of pulmonary inflammation induced by MHV-1 infection in susceptible (A/J, C3H/HeJ, and BALB/c) and resistant (C57BL/6) strains of mice. Virus replication and distribution did not correlate with the relative susceptibilities of A/J, BALB/c, C3H/HeJ, and C57BL/6 mice. In order to further define the role of the host genetic background in influencing susceptibility to MHV-1-induced disease, we examined 14 different inbred mouse strains. BALB.B and BALB/c mice exhibited MHV-1-induced weight loss, whereas all other strains of H-2(b) and H-2(d) mice did not show any signs of disease following MHV-1 infection. H-2(k) mice demonstrated moderate susceptibility, with C3H/HeJ mice exhibiting the most severe disease. C3H/HeJ mice harbor a natural mutation in the gene that encodes Toll-like receptor 4 (TLR4) that disrupts TLR4 signaling. C3H/HeJ mice exhibit enhanced morbidity and mortality following i.n. MHV-1 infection compared to wild-type C3H/HeN mice. Our results indicate that TLR4 plays an important role in respiratory CoV pathogenesis. PMID- 19553338 TI - Arginine methylation of the ICP27 RGG box regulates the functional interactions of ICP27 with SRPK1 and Aly/REF during herpes simplex virus 1 infection. AB - The herpes simplex virus 1 protein ICP27 is methylated on arginine residues within an RGG box, and arginine methylation regulates ICP27 export to the cytoplasm. Arginine methylation can regulate protein-protein interactions; therefore, we examined the effect of hypomethylation on ICP27's interactions with cellular proteins SRPK1 and Aly/REF, which bind to ICP27 through the RGG box region. During infections with viral mutants containing lysine substitutions or the methylation inhibitor adenosine dialdehyde, the interaction of ICP27 with SRPK1 and Aly/REF was decreased, as determined by coimmunoprecipitation and colocalization studies, indicating that ICP27 RGG box methylation regulates interaction with these proteins. PMID- 19553340 TI - Identification of unusual E6 and E7 proteins within avian papillomaviruses: cellular localization, biophysical characterization, and phylogenetic analysis. AB - Papillomaviruses (PVs) are a large family of small DNA viruses infecting mammals, reptiles, and birds. PV infection induces cell proliferation that may lead to the formation of orogenital or skin tumors. PV-induced cell proliferation has been related mainly to the expression of two small oncoproteins, E6 and E7. In mammalian PVs, E6 contains two 70-residue zinc-binding repeats, whereas E7 consists of a natively unfolded N-terminal region followed by a zinc-binding domain which folds as an obligate homodimer. Here, we show that both the novel francolin bird PV Francolinus leucoscepus PV type 1 (FlPV-1) and the chaffinch bird PV Fringilla coelebs PV contain unusual E6 and E7 proteins. The avian E7 proteins contain an extended unfolded N terminus and a zinc-binding domain of reduced size, whereas the avian E6 proteins consist of a single zinc-binding domain. A comparable single-domain E6 protein may have existed in a common ancestor of mammalian and avian PVs. Mammalian E6 C-terminal domains are phylogenetically related to those of single-domain avian E6, whereas mammalian E6 N-terminal domains seem to have emerged by duplication and subsequently diverged from the original ancestral domain. In avian and mammalian cells, both FlPV-1 E6 and FlPV-1 E7 were evenly expressed in the cytoplasm and the nucleus. Finally, samples of full-length FlPV-1 E6 and the FlPV-1 E7 C-terminal zinc-binding domain were prepared for biophysical analysis. Both constructs were highly soluble and well folded, according to nuclear magnetic resonance spectroscopy measurements. PMID- 19553339 TI - Matrix metalloprotease inhibitors restore impaired NK cell-mediated antibody dependent cellular cytotoxicity in human immunodeficiency virus type 1 infection. AB - Increasing evidence suggests that NK cells not only are critical in the initial host defense against pathogens but also may contribute to continued protection from human immunodeficiency virus type 1 (HIV-1) disease progression. NK cell cytolysis can be induced directly through diverse receptor families or can be induced indirectly through Fc receptors by antibodies mediating antibody dependent cellular cytotoxicity (ADCC). ADCC has been implicated in both protection from simian immunodeficiency virus infection and slower progression of HIV-1 disease. ADCC activity declines with advancing infection, and yet the underlying mechanism for this dysfunction has not been defined, nor has it been determined whether the activity can be reconstituted. Here we demonstrate that NK cell-mediated ADCC is severely compromised in chronic HIV infection. The potency of ADCC function was directly correlated with baseline Fc gammaRIIIa receptor (CD16) expression on NK cells. CD16 expression was negatively influenced by elevated expression of a group of enzymes, the matrix metalloproteinases (MMPs), normally involved in tissue/receptor remodeling. Inhibition of MMPs resulted in increased CD16 expression and augmented ADCC activity in response to antibody coated target cells. These data suggest that MMP inhibitors may improve NK cell mediated ADCC, which may provide subjects with an opportunity to harness the cytolytic power of NK cells through naturally occurring nonneutralizing HIV specific antibodies. PMID- 19553341 TI - Low endocytic pH and capsid protein autocleavage are critical components of Flock House virus cell entry. AB - The process by which nonenveloped viruses cross cell membranes during host cell entry remains poorly defined; however, common themes are emerging. Here, we use correlated in vivo and in vitro studies to understand the mechanism of Flock House virus (FHV) entry and membrane penetration. We demonstrate that low endocytic pH is required for FHV infection, that exposure to acidic pH promotes FHV-mediated disruption of model membranes (liposomes), and particles exposed to low pH in vitro exhibit increased hydrophobicity. In addition, FHV particles perturbed by heating displayed a marked increase in liposome disruption, indicating that membrane-active regions of the capsid are exposed or released under these conditions. We also provide evidence that autoproteolytic cleavage, to generate the lipophilic gamma peptide (4.4 kDa), is required for membrane penetration. Mutant, cleavage-defective particles failed to mediate liposome lysis, regardless of pH or heat treatment, suggesting that these particles are not able to expose or release the requisite membrane-active regions of the capsid, namely, the gamma peptides. Based on these results, we propose an updated model for FHV entry in which (i) the virus enters the host cell by endocytosis, (ii) low pH within the endocytic pathway triggers the irreversible exposure or release of gamma peptides from the virus particle, and (iii) the exposed/released gamma peptides disrupt the endosomal membrane, facilitating translocation of viral RNA into the cytoplasm. PMID- 19553342 TI - Kaposi's sarcoma-associated herpesvirus protein LANA2 disrupts PML oncogenic domains and inhibits PML-mediated transcriptional repression of the survivin gene. AB - Infection by herpesviruses causes a dramatic disturbance of PML oncogenic domains (PODs) that has been suggested to be essential for viral lytic replication. Several proteins from Kaposi's sarcoma-associated herpesvirus (KSHV) have been tested as putative POD-disrupting factors with negative results. Here, we show that LANA2, a viral protein that is absolutely required for the viability and proliferation of KSHV-infected primary effusion lymphoma (PEL) cells, increases the levels of SUMO2-ubiquitin-modified PML and induces the disruption of PODs by a proteasome-mediated mechanism. In addition, we demonstrate that this disruption is largely dependent on both the integrity of a SUMO interaction motif in LANA2 and the lysine 160 from PML. Moreover, silencing of LANA2 expression in PEL cells by RNA interference led to an increase in the PML levels. Finally, we demonstrate that LANA2 relieves PML-mediated transcriptional repression of survivin, a protein that directly contributes to malignant progression of PEL. This represents the first example of inactivation of these important antiviral structures by KSHV. PMID- 19553343 TI - Fluorescently labeled ribosomes as a tool for analyzing antibiotic binding. AB - Measuring the binding of antibiotics and other small-molecular-weight ligands to the 2.5 MDa ribosome often presents formidable challenges. Here, we describe a general method for studying binding of ligands to ribosomes that carry a site specific fluorescent label covalently attached to one of the ribosomal proteins. As a proof of principle, an environment-sensitive fluorescent group was placed at several specific sites within the ribosomal protein S12. Small ribosomal subunits were reconstituted from native 16S rRNA, individually purified small subunit proteins, and fluorescently labeled S12. The fluorescence characteristics of the reconstituted subunits were affected by several antibiotics, including streptomycin and neomycin, which bind in the vicinity of protein S12. The equilibrium dissociation constants of the drugs obtained using a conventional fluorometer were in good agreement with those observed using previously published methods and with measurements based on the use of radiolabeled streptomycin. The newly developed method is rapid and sensitive, and can be used for determining thermodynamic and kinetic binding characteristics of antibiotics and other small ribosomal ligands. The method can readily be adapted for use in high-throughput screening assays. PMID- 19553344 TI - Deep sequencing identifies new and regulated microRNAs in Schmidtea mediterranea. AB - MicroRNAs (miRNAs) play important roles in directing the differentiation of cells down a variety of cell lineage pathways. The planarian Schmidtea mediterranea can regenerate all lost body tissue after amputation due to a population of pluripotent somatic stem cells called neoblasts, and is therefore an excellent model organism to study the roles of miRNAs in stem cell function. Here, we use a combination of deep sequencing and bioinformatics to discover 66 new miRNAs in S. mediterranea. We also identify 21 miRNAs that are specifically expressed in either sexual or asexual animals. Finally, we identified five miRNAs whose expression is sensitive to gamma-irradiation, suggesting they are expressed in neoblasts or early neoblast progeny. Together, these results increase the known repertoire of S. mediterranea miRNAs and identify numerous regulated miRNAs that may play important roles in regeneration, homeostasis, neoblast function, and reproduction. PMID- 19553345 TI - Regulatory element identification in subsets of transcripts: comparison and integration of current computational methods. AB - Regulatory elements in mRNA play an often pivotal role in post-transcriptional regulation of gene expression. However, a systematic approach to efficiently identify putative regulatory elements from sets of post-transcriptionally coregulated genes is lacking, hampering studies of coregulation mechanisms. Although there are several analytical methods that can be used to detect conserved mRNA regulatory elements in a set of transcripts, there has been no systematic study of how well any of these methods perform individually or as a group. We therefore compared how well three algorithms, each based on a different principle (enumeration, optimization, or structure/sequence profiles), can identify elements in unaligned untranslated sequence regions. Two algorithms were originally designed to detect transcription factor binding sites, Weeder and BioProspector; and one was designed to detect RNA elements conserved in structure, RNAProfile. Three types of elements were examined: (1) elements conserved in both primary sequence and secondary structure; (2) elements conserved only in primary sequence; and (3) microRNA targets. Our results indicate that all methods can uniquely identify certain known RNA elements, and therefore, integrating the output from all algorithms leads to the most complete identification of elements. We therefore developed an approach to integrate results and guide selection of candidate elements from several algorithms presented as a web service (https://dbw.msi.umn.edu:8443/recit). These findings together with the approach for integration can be used to identify candidate elements from genome-wide post-transcriptional profiling data sets. PMID- 19553346 TI - Effects of nitric oxide and antioxidants on advanced glycation end products induced hypertrophic growth in human renal tubular cells. AB - The accumulation of advanced glycation end products (AGE) is a key mediator of renal tubular hypertrophy in diabetic nephropathy (DN). Reactive oxygen species and nitric oxide (NO) were involved in the progression of DN. In this study, the molecular mechanisms of NO and antioxidants responsible for inhibition of AGE induced renal tubular hypertrophy were examined. We found that AGE (but not nonglycated bovine serum albumin) significantly suppressed the NO/cGMP/PKG signaling in human renal proximal tubular cells. NO donors S-nitroso-N acetylpenicillamine (SNAP)/sodium nitroprusside (SNP) and antioxidants N acetylcysteine (NAC)/taurine treatments significantly attenuated AGE-inhibited NO production, cGMP synthesis, and inducible NO synthase/cGMP-dependent protein kinase (PKG) activation. Moreover, AGE-induced extracellular signal-regulated kinase/c-Jun N-terminal kinase/p38 mitogen-activated protein kinase activation was markedly blocked by antireceptor for AGE (RAGE), SNAP, SNP, NAC, and taurine. The abilities of NO and antioxidants to inhibit AGE/RAGE-induced hypertrophic growth were verified by the observation that SNAP, SNP, NAC, and taurine inhibited fibronectin, p21(Waf1/Cip1), and RAGE expression. Therefore, antioxidants significantly attenuated AGE/RAGE-enhanced cellular hypertrophy partly through induction of the NO/cGMP/PKG signaling. PMID- 19553347 TI - Human immunodeficiency virus downregulates podocyte apoE expression. AB - Apolipoprotein E (apoE) has been demonstrated to play an important role in providing protection against mesangial cell injury. In the present study, we evaluated the role of apoE and its associated downstream effects in human immunodeficiency virus (HIV)-associated nephropathy (HIVAN). Control (n = 6) and age- and sex-matched HIV-1 transgenic mice (Tg26, n = 6) were evaluated for their renal cortical expression of apoE. Renal tissue from Tg26 mice not only showed decreased apoE expression but also displayed downregulation of perlecan mRNA expression. In in vitro studies, conditionally immortalized human podocytes (CIHPs) were transduced with either NL4-3HIV (an HIV-1 construct lacking gag and pol, used for the development of Tg26 mouse model; NL4-3/CIHP) or empty vector (EV/CIHP); NL4-3/CIHPs and EV/CIHPs were studied for apoE mRNA expression. NL4 3/CIHPs showed reduction in apoE expression compared with EV/CIHPs. To evaluate the role of HIV-1 genes in the modulation of apoE expression, conditionally immortalized mouse podocytes (CIMPs) were transduced with individual HIV-1 gene constructs. Only nef-transduced CIMPs showed a decrease in apoE expression. To confirm this effect of nef in CIHPs, microarray analysis was performed in stable colonies of nef/CIHPs and EV/CIHPs. nef/CIHPs showed a 60% decrease in apoE and a 90% reduction in heparan sulfate mRNA expression. Moreover, nef transgenic mice showed a decrease in renal tissue expression of both apoE and perlecan. Both Tg26 and nef transgenic mice also showed areas of mesangial cell proliferation. These findings suggest that HIV-1-induced reduction in podocyte apoE expression and associated downregulation of podocyte perlecan might be contributing to mesangial cell (MC) phenotype in HIVAN. PMID- 19553348 TI - Cyclophilin D gene ablation protects mice from ischemic renal injury. AB - Increased oxidative stress and intracellular calcium levels and mitochondrial overloading of calcium during ischemic renal injury (IRI) favor mitochondrial membrane permeability transition pore (MPTP) opening and subsequent necrotic cell death. Cyclophilin D (CypD) is an essential component of MPTP, and recent findings implicate its role in necrotic, but not apoptotic, cell death. To evaluate the role of CypD following IRI, we tested the hypothesis that CypD gene ablation protects mice from IRI. Renal function as assessed by plasma levels of both creatinine and blood urea nitrogen was significantly reduced in CypD knockout (CypD(-/-)) mice compared with wild-type mice during the 5-day post ischemia period. Erythrocyte trapping, tubular cell necrosis, tubular dilatation, and neutrophil infiltration were significantly decreased in CypD(-/-) mice. To define the mechanisms by which CypD deficiency protect the kidneys, an in vitro model of IRI was employed. Inhibition of CypD using cyclosporin A in oxidant injured cultured proximal tubular cells (PTC) prevented mitochondrial membrane depolarization, reduced LDH release, ATP depletion and necrotic cell death. Similarly, oxidant-injured CypD(-/-) PTC primary cultures were protected from cytotoxicity and necrosis. To conclude, CypD gene ablation offers both functional and morphological protection in mice following IRI by decreasing necrotic cell death possibly via inhibition of MPTP and ATP depletion. PMID- 19553349 TI - Soluble epoxide hydrolase gene deletion attenuates renal injury and inflammation with DOCA-salt hypertension. AB - Inhibition of soluble epoxide hydrolase (sEH) has been shown to be renal protective in rat models of salt-sensitive hypertension. Here, we hypothesize that targeted disruption of the sEH gene (Ephx2) prevents both renal inflammation and injury in deoxycorticosterone acetate plus high salt (DOCA-salt) hypertensive mice. Mean arterial blood pressure (MAP) increased significantly in the DOCA-salt groups, and MAP was lower in Ephx2-/- DOCA-salt (129 +/- 3 mmHg) compared with wild-type (WT) DOCA-salt (145 +/- 2 mmHg) mice. Following 21 days of treatment, WT DOCA-salt urinary MCP-1 excretion increased from control and was attenuated in the Ephx2-/- DOCA-salt group. Macrophage infiltration was reduced in Ephx2-/- DOCA-salt compared with WT DOCA-salt mice. Albuminuria increased in WT DOCA-salt (278 +/- 55 microg/day) compared with control (17 +/- 1 microg/day) and was blunted in the Ephx2-/- DOCA-salt mice (97 +/- 23 microg/day). Glomerular nephrin expression demonstrated an inverse relationship with albuminuria. Nephrin immunofluorescence was greater in the Ephx2-/- DOCA-salt group (3.4 +/- 0.3 RFU) compared with WT DOCA-salt group (1.1 +/- 0.07 RFU). Reduction in renal inflammation and injury was also seen in WT DOCA-salt mice treated with a sEH inhibitor {trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid; tAUCB}, demonstrating that the C-terminal hydrolase domain of the sEH enzyme is responsible for renal protection with DOCA-salt hypertension. These data demonstrate that Ephx2 gene deletion decreases blood pressure, attenuates renal inflammation, and ameliorates glomerular injury in DOCA-salt hypertension. PMID- 19553350 TI - Histone deacetylase-2 is a key regulator of diabetes- and transforming growth factor-beta1-induced renal injury. AB - Excessive accumulation of extracellular matrix (ECM) in the kidneys and epithelial-to-mesenchymal transition (EMT) of renal tubular epithelial cells contributes to the renal fibrosis that is associated with diabetic nephropathy. Histone deacetylase (HDAC) determines the acetylation status of histones and thereby controls the regulation of gene expression. This study examined the effect of HDAC inhibition on renal fibrosis induced by diabetes or transforming growth factor (TGF)-beta1 and determined the role of reactive oxygen species (ROS) as mediators of HDAC activation. In streptozotocin (STZ)-induced diabetic kidneys and TGF-beta1-treated normal rat kidney tubular epithelial cells (NRK52 E), we found that trichostatin A, a nonselective HDAC inhibitor, decreased mRNA and protein expressions of ECM components and prevented EMT. Valproic acid and class I-selective HDAC inhibitor SK-7041 also showed similar effects in NRK52-E cells. Among the six HDACs tested (HDAC-1 through -5 and HDAC-8), HDAC-2 activity significantly increased in the kidneys of STZ-induced diabetic rats and db/db mice and TGF-beta1-treated NRK52-E cells. Levels of mRNA expression of fibronectin and alpha-smooth muscle actin were decreased, whereas E-cadherin mRNA was increased when HDAC-2 was knocked down using RNA interference in NRK52-E cells. Interestingly, hydrogen peroxide increased HDAC-2 activity, and the treatment with an antioxidant, N-acetylcysteine, almost completely reduced TGF beta1-induced activation of HDAC-2. These findings suggest that HDAC-2 plays an important role in the development of ECM accumulation and EMT in diabetic kidney and that ROS mediate TGF-beta1-induced activation of HDAC-2. PMID- 19553352 TI - Healthcare waste management - are we changing? Yes we are - slowly! PMID- 19553351 TI - Hsp27 inhibits sublethal, Src-mediated renal epithelial cell injury. AB - Disruption of cell contact sites in renal epithelial cells contributes to organ dysfunction after ischemia. We hypothesized that heat shock protein 27 (Hsp27), a known cytoprotectant protein, preserves cell architecture and cell contact site function during ischemic stress. To test this hypothesis, renal epithelial cells were subjected to transient ATP depletion, an in vitro model of ischemia reperfusion injury. Compared with control, selective Hsp27 overexpression significantly preserved cell-cell junction function during metabolic stress as evidenced by reduced stress-mediated redistribution of the adherens junction protein E-cadherin, higher transepithelial electrical resistance, and lower unidirectional flux of lucifer yellow. Hsp27 overexpression also preserved paxillin staining within focal adhesion complexes and significantly decreased cell detachment during stress. Surprisingly, Hsp27, an F-actin-capping protein, only minimally reduced stress induced actin cytoskeleton collapse. In contrast to Hsp27 overexpression, siRNA-mediated knockdown had the opposite effect on these parameters. Since ischemia activates c-Src, a tyrosine kinase that disrupts both cell-cell and cell-substrate interactions, the relationship between Hsp27 and c Src was examined. Although Hsp27 and c-Src did not coimmunoprecipitate and Hsp27 overexpression failed to inhibit whole cell c-Src activation during injury, manipulation of Hsp27 altered active c-Src accumulation at cell contact sites. Specifically, Hsp27 overexpression reduced, whereas Hsp27 knockdown increased active p-(416)Src detected at contact sites in intact cells as well as in a purified cell membrane fraction. Together, this evidence shows that Hsp27 overexpression prevents sublethal REC injury at cell contact sites possibly by a c-Src-dependent mechanism. Further exploration of the biochemical link between Hsp27 and c-Src could yield therapeutic interventions for ameliorating ischemic renal cell injury and organ dysfunction. PMID- 19553353 TI - The challenge of medical waste management: a case study in northwest Iran-Tabriz. AB - This article presents the results of a descriptive cross-sectional study on medical waste management in Tabriz (Iran's fourth largest city). The study was conducted in 10 of 25 active hospitals of the city in the summer of 2007. The methodology of the present study was based on data collected from hospitals through a checklist, site visits (observation), and quantity analysis by weight. The results indicated that more than 13.59 tonnes day(-1) of total medical waste and 4.06 tonnes day(-1) of hazardous-infectious medical waste are generated by the active hospitals of the city. Currently, there are no practical instructions, or suitable supervision on different levels of waste management. The health authorities and hospital managers do not accept sufficient responsibility for the medical waste due to financial problems and the lack of awareness regarding the hazards of medical waste. Segregation and minimization of waste are not carried out correctly in any of the hospitals. The use of protective measures by staff and temporary storage areas was not in agreement with standards in 70 and 60% of the hospitals in the present study, respectively. About 50% of the hospitals had been equipped with an incinerator, but all but one (10%) of them had been phased out due to operation and maintenance problems, air pollution, etc. Almost all of the hospitals have a waste management officer, but there is not an effective training programme for the staff. Infectious-hazardous medical waste is mixed with general waste, and it is disposed of in a municipal waste landfill, which is an unsanitary dumpsite. Illegal segregation and recycling of medical waste is carried out at the final disposal site; therefore, there are concerns about environmental pollution and the transmission of infectious diseases. It is proposed that, through the allocation of increased budgets, implementation of integrated segregation, minimization of waste, and creation of a training programme in the hospitals, the quantity of medical waste would be decreased (by about 70.11%). Considering the previous unsuccessful experience of on-site incineration in Tabriz (and in Iran's other large cites), an amendment should be made to Iran's current hazardous waste regulations to have infectious-hazardous waste sent to a central off-site autoclave or incinerator for treatment. The off site autoclave would have some advantages, such as decreased air pollution. Of course, some health officials oppose this plan. To test this plan and receive the official's approval, a central off-site autoclave can be put into practice as a pilot. PMID- 19553354 TI - Delegation guided by school nursing values: comprehensive knowledge, trust, and empowerment. AB - As health care institutions in the United States respond to shrinking budgets and nursing shortages by increasing the use of unlicensed assistive personnel (UAP), school nursing practice is changing from providing direct care to supervising activities delegated to UAP. Therefore, delegation is a critical area of concern for school nurses. The purpose of this qualitative research study was to explore values guiding the delegation of health care tasks to UAP in school settings from the perspective of the school nurse. An inquiry focus group was conducted with 64 Florida school nurses. Values guiding delegation were comprehensive knowledge, trust, and empowerment. These values provided a framework for guiding the delegation process. PMID- 19553355 TI - Quantification of oocyte-specific transcripts in follicle-enclosed oocytes during antral development and maturation in vitro. AB - Oocyte cytoplasmic maturation is influenced by the quantity of synthesized RNA and proteins accumulated and stored during growth. Transcriptional repression and degradation of transcripts occur during oocyte nuclear maturation, and prolonged transcriptional arrest might compromise RNA stores for early development. RNA quantification of key genes in oocytes might be valuable when setting up in vitro cultures that lack the normal hormonal interplay found in vivo. This study quantifies gene expression levels in relation to follicle culture time and time of oocyte maturation in a mouse model. RNA levels of Gdf-9, Bmp-15, Mater, Zar-1, Npm-2 and Fgf-8 were measured in germinal vesicle oocytes along fixed times during in vitro follicle development. For all genes, the highest mRNA levels were detected in oocytes in the pre-antral follicle stage. Antrum formation was associated with a progressive shutdown in transcription leading to mRNA values lower than those in vivo preovulatory oocytes by extending period of in vitro culture. In contrast to in vitro-matured oocytes, the in vivo oocytes from 22- and 29-day-old prepubertal animals obtained after pregnant mare's serum gonadotrophin and human chorionic gonadotrophin priming did not down-regulate transcripts upon maturation stimulus except for Mater. These findings show that oocyte gene expression patterns under in vitro conditions can, at certain times, mimic what is reported to occur under in vivo conditions. Moreover, they also show that meiotically competent oocytes kept in a prolonged transcriptionally inactive stage express altered levels of key transcripts compared with in vivo in both immature and mature oocytes. PMID- 19553356 TI - Rank-based estimation in the {ell}1-regularized partly linear model for censored outcomes with application to integrated analyses of clinical predictors and gene expression data. AB - We consider estimation and variable selection in the partial linear model for censored data. The partial linear model for censored data is a direct extension of the accelerated failure time model, the latter of which is a very important alternative model to the proportional hazards model. We extend rank-based lasso type estimators to a model that may contain nonlinear effects. Variable selection in such partial linear model has direct application to high-dimensional survival analyses that attempt to adjust for clinical predictors. In the microarray setting, previous methods can adjust for other clinical predictors by assuming that clinical and gene expression data enter the model linearly in the same fashion. Here, we select important variables after adjusting for prognostic clinical variables but the clinical effects are assumed nonlinear. Our estimator is based on stratification and can be extended naturally to account for multiple nonlinear effects. We illustrate the utility of our method through simulation studies and application to the Wisconsin prognostic breast cancer data set. PMID- 19553357 TI - Sociodemographic and lifestyle risk factors for incident dementia and cognitive decline in the HYVET. AB - INTRODUCTION: previous studies have suggested that smoking, living alone and having a high body mass index may increase risk of developing dementia whereas a normal body mass index, having received education and moderate alcohol consumption may decrease risk. Dementia risk also increases with age and is thought to be higher in hypertensives. METHOD: we used data collected in the Hypertension in the Very Elderly Trial (HYVET), and cognitive function was assessed using the Mini-Mental State Examination (MMSE) at baseline and annually. Participants with a fall in MMSE to <24 or with a fall of 3 points in any 1 year were investigated further. The association of baseline sociodemographic, medical and lifestyle factors with incident dementia or decline in MMSE scores was assessed by regression models. RESULTS: incident dementia occurred in 263 of 3,336 participants over a mean follow-up of 2 years. In multivariate analyses, being underweight, BMI < 18.5 (HR 1.90, 95% CI 1.06-3.39) or obese, BMI >30 (HR 1.84, 95% CI 1.24-2.72), increased risk of incident dementia as did piracetam use (HR 2.72, 95% CI 1.60-4.63). Receiving formal education was associated with a reduced risk (HR 0.59, 95% CI 0.45-0.78). There was no association with smoking, alcohol and gender. Similar results were found when examining mean annual change in the MMSE score. DISCUSSION: our results for BMI and education agree with those from other studies. The increased risk associated with piracetam may reflect awareness of memory problems before any diagnosis of dementia has been made. Trial participants may be healthier than the general population and further studies in the general population are required. PMID- 19553358 TI - A comparison of four tests of cognition as predictors of inability to perform spirometry in old age. AB - BACKGROUND: previous studies have shown that a Mini Mental State Examination (MMSE) score of <24/30 and inability to copy intersecting pentagons (IP) predicts inability to perform spirometry. We hypothesised that clock drawing tests (CLOX 1 and 2), being validated tests of cognitive executive function, might predict spirometry performance with a higher sensitivity and specificity than the MMSE or IP. METHODS: we studied 113 (84 females) spirometry-naive inpatients, mean age of 84 years (range 74-97). All performed the MMSE, IP, CLOX 1 and 2 and then attempted to perform assisted spirometry to the American Thoracic Society/European Respiratory Society standard. RESULTS: of 113, 49 met the criteria for adequate spirometry. Using normative thresholds for probable impairment, inability to perform spirometry was predicted by MMSE <24/30 with a sensitivity of 81% and specificity of 90% (P<0.0000); by inability to copy IP with a sensitivity of 92% and specificity of 100% (P<0.0000); by CLOX1 <10/15 with a sensitivity of 81% and specificity of 49% (P<0.001); and by CLOX2 <12/15 with a sensitivity of 63% and specificity of 65% (P<0.001). CONCLUSION: CLOX tests did not perform better than MMSE and IP to identify subjects unlikely to be able to perform spirometry. Achieving assisted spirometry from the naive state in old age might be more determined by global cognitive function and ideo-motor praxis than by executive control function. PMID- 19553359 TI - Cardiovascular molecular imaging: an overview. AB - Molecular imaging is non-invasive visualization and measurement of biological processes at the molecular and cellular level within a living organism. This review provides a description of the various molecular imaging techniques for imaging cardiovascular targets and their potential clinical implications. Molecular imaging has relied mainly on nuclear imaging, but advances in nanoparticle probe development have made magnetic resonance imaging and ultrasound as emerging, radiation-free alternatives. Targeted imaging of vascular inflammation or thrombosis may allow improved risk assessment of atherosclerosis by detecting plaques at high risk of acute complications. Imaging probes detecting myocardial apoptosis, metabolic alterations, injury to extracellular matrix, angiogenesis, or innervation may provide tools for assessing risk of arrhythmias and left ventricular remodelling associated with progressive cardiac dysfunction and heart failure. Although clinical experience remains limited, careful evaluation of safety as well as validation of diagnostic and prognostic value of these techniques in clinical trials is still needed. PMID- 19553360 TI - Compeer friends: a qualitative study of a volunteer friendship programme for people with serious mental illness. AB - BACKGROUND: People with serious mental illness (SMI) experience numerous barriers to developing and maintaining friendships. AIMS: To explore the benefits and drawbacks of an intentional friendship programme (Compeer, Inc), which develops new social relationships for people with SMI by matching them in one-to-one relationships with community volunteers for weekly social activities. METHODS: Twenty clients and volunteers, in Compeer friendships for different lengths of time, participated in individual semi-structured qualitative interviews. Several volunteers were themselves current or former consumers of mental health services. RESULTS: Participants reported numerous benefits to participating in Compeer. Clients and volunteers spoke enthusiastically about the benefits of gaining a friend. Many intentional relationships deepened over several years into mutually beneficial friendships. Most clients became more outgoing, sociable and active, with increased self-esteem, self-worth and self-confidence. Volunteers who had experienced mental illness themselves provided unique added benefits to the relationship. Drawbacks were minimal and financial and other costs to volunteers were low. CONCLUSIONS: Intentional friendships can be a potent yet cost-effective way to help people with SMI develop social skills, expand their social networks, and improve their quality of life. However, because relationships take several years to develop, quantitative evaluations using short follow-up periods may underestimate programme effectiveness. PMID- 19553361 TI - International migration of partner, autonomy and depressive symptoms among women from a mexican rural area. AB - BACKGROUND: The emigration of Mexicans to the USA has increased in the last decades, and little is known about the effect of this on the mental health of those who stay behind. AIMS: To evaluate the association of emigration of husband and depressive symptoms (DS) among women who stay in Mexico. We also tested the hypothesis that the husband's migration would increase the woman's autonomy, which in turn would decrease DS. METHODS: A survey was conducted in a rural area in Mexico. Participants (n = 418) were selected through probabilistic sampling in three stages: localities, households and individuals. DS were evaluated using the Centre for Epidemiological Studies-Depression (CES-D) scale. RESULTS: Having a partner in the USA was associated with higher odds of scoring above the cut-off point in CES-D (OR 3.77, 95% CI 1.92-7.43). Economic autonomy was also associated with DS (OR 1.45, 95% CI 1.04-2.02). CONCLUSION: Migration of husband was associated with DS among women. The construct of autonomy and its operational definition should be further explored. PMID- 19553362 TI - Effects of obesity and obesity-induced stress on depressive symptoms in Korean elementary school children. AB - BACKGROUND: Obesity is becoming prevalent in Korean children. Because body image is becoming increasingly important, it is likely that obesity-induced stress has a significant effect on childhood depression. AIMS: To examine the correlation between obesity-induced stress and depressive symptoms in Korean elementary school students. METHODS: The study participants were 2,305 elementary school children and their parents in the districts of Jeju-si, Seogwipo-si, Namjeju-gun and Bukjeju-gun on Jeju Island, Korea, who completed questionnaires involving demographic information, an obesity-induced stress scale and the Korean form of Kovacs' Children's Depression Inventory (CDI) from September to December 2006. RESULTS: After controlling for significant independent variables that are wellknown correlates of depressive symptoms in children (e.g. age, gender, residence, family monthly income, obesity status of both parents, family history of chronic illness, and time spent with mother), obesity-induced stress had an odds ratio of 1.128 (95% CI 1.111-1.146). CONCLUSIONS: Reducing the prevalence of depressive symptoms in elementary school children in Jeju Island will require special attention, particularly the development of coping strategies to resolve obesity-induced stress in various areas including school, family and society. PMID- 19553364 TI - Factors, process and outcomes of recovery from psychiatric disability: the unity model. AB - BACKGROUND: Despite the fruitful findings on related issues of recovery in the West, some researchers have called for more studies on the factors that facilitate recovery and international literature on recovery to be made available. Moreover, to date, a united model that integrates outcome, component process and contextual factors of recovery has not yet been developed. Thus, this study explored the recovery experiences of persons with psychiatric disabilities (hereinafter called consumers) in Taiwan and extracted the key facilitators for developing a preliminary unity theory of recovery. MATERIAL: In-depth qualitative interviews of 15 consumers in recovery and their caregivers were held. Over a research period of two years, consumers were interviewed twice. The dialogue of each interview was transcribed into text and a narrative summary of the storyline for each participant was also prepared. DISCUSSION: For most consumers, the journey of recovery was an incremental process of progress, yet few of them mentioned a turning point and its significant change on their life. Regaining social roles seemed to be a necessary but not sufficient outcome indicator for recovery. While symptom remission, mental strength and parental support were the cornerstones for recovery, the treatment model and professionals got the credit for it too. CONCLUSION: The recovery process occurs within a complex context of various stages and multi-facilitators. The forces of three cornerstones, essential components and contextual facilitators all influx into the river of recovery and emerge as one united mechanism that supports the consumer's spiral progress through the journey of striving for autonomy. PMID- 19553363 TI - Social relationships as a decisive factor in recovering from severe mental illness. AB - BACKGROUND: Recovery research often describes recovery from mental illness as a complex individual process. In this article a social perspective on recovery is developed. AIMS: To ascertain which factors people regard as decisive to their own recovery and what makes them beneficial. METHODS: In-depth interviews were conducted with 58 persons in Sweden who had recovered from severe mental illness. Interviews were qualitatively analyzed using grounded theory. RESULTS: Three dimensions of contributing recovery factors were identified. Social relationships emerged as the core category throughout these dimensions. CONCLUSIONS: The results show that recovery processes are social processes in which social relationships play a key role. PMID- 19553365 TI - Knowledge about schizophrenia and attitudes towards people with schizophrenia in Greece. AB - BACKGROUND: This study was the first national survey ever conducted in Greece aiming to monitor the level of stigma Greeks hold against people with schizophrenia (PWS). AIM: To investigate sources, degree of knowledge and attitudes towards PWS in Greece. METHOD: A cross-sectional nationwide survey was conducted by face-to-face household interviews ( n = 1,199, aged 15 years and over). RESULTS: Television was the main source of information (65.9%). Negative portrayals of PWS were recalled by 60.5%. Only 27.7% attributed schizophrenia to a combination of psychosocial, genetic and environmental factors. Respondents believed that PWS are dangerous (74.6%), have split personalities (81.3%) and cannot work (83.2%). Regarding attitudes, a negative relationship between closeness and social distance was observed. Most respondents (92.1%) would not marry someone with schizophrenia, half (50.5%) would be disturbed by working with PWS and one third (32.9%) would feel afraid to start a conversation. Urban residence and higher education were generally associated with better knowledge and more positive attitudes towards PWS. CONCLUSIONS: Knowledge about schizophrenia in Greece is poor. The Greek public has stigmatizing attitudes towards PWS. Educational interventions should especially target rural and semi urban residents of a lower educational level. The role of television can be crucial. PMID- 19553366 TI - Psychological help-seeking in homeless adolescents. AB - BACKGROUND: Homeless young people in the UK are a vulnerable group: their levels of psychological problems, physical ill health and violent victimisation exceed those of their housed counterparts. AIMS: This qualitative study aimed to examine homeless young people's views about seeking psychological help for their problems. METHOD: Semi-structured interviews were undertaken with 16 homeless young people at an emergency hostel. A thematic analysis and an analysis of the participants' narratives were undertaken. RESULTS: A sense of hurt and anger at the perceived betrayal by their families and society made many participants reluctant to seek help and to trust help offered by others, and accordingly they placed a high value on self-sufficiency. Many said that they would, however, seek help from people whom they perceived to be genuine, caring, trustworthy, empathic and capable of containing their distress. CONCLUSION: Mental health professionals providing clinical services for this underserved population need to be aware of the importance of rejection and abandonment issues, along with the consequent hurt, anger and mistrust. PMID- 19553367 TI - Evolutionary origin and functions of retrogene introns. AB - Retroposed genes (retrogenes) originate via the reverse transcription of mature messenger RNAs from parental source genes and are therefore usually devoid of introns. Here, we characterize a particular set of mammalian retrogenes that acquired introns upon their emergence and thus represent rare cases of intron gain in mammals. We find that although a few retrogenes evolved introns in their coding or 3' untranslated regions (untranslated region, UTR), most introns originated together with untranslated exons in the 5' flanking regions of the retrogene insertion site. They emerged either de novo or through fusions with 5' UTR exons of host genes into which the retrogenes inserted. Generally, retrogenes with introns display high transcription levels and show broader spatial expression patterns than other retrogenes. Our experimental expression analyses of individual intron-containing retrogenes show that 5' UTR introns may indeed promote higher expression levels, at least in part through encoded regulatory elements. By contrast, 3' UTR introns may lead to downregulation of expression levels via nonsense-mediated decay mechanisms. Notably, the majority of retrogenes with introns in their 5' flanks depend on distant, sometimes bidirectional CpG dinucleotide-enriched promoters for their expression that may be recruited from other genes in the genomic vicinity. We thus propose a scenario where the acquisition of new 5' exon-intron structures was directly linked to the recruitment of distant promoters by these retrogenes, a process potentially facilitated by the presence of proto-splice sites in the genomic vicinity of retrogene insertion sites. Thus, the primary role and selective benefit of new 5' introns (and UTR exons) was probably initially to span the often substantial distances to potent CpG promoters driving retrogene transcription. Later in evolution, these introns then obtained additional regulatory roles in fine tuning retrogene expression levels. Our study provides novel insights regarding mechanisms underlying the origin of new introns, the evolutionary relevance of intron gain, and the origin of new gene promoters. PMID- 19553368 TI - Identification of leaf proteins differentially accumulated during cold acclimation between Festuca pratensis plants with distinct levels of frost tolerance. AB - Festuca pratensis (meadow fescue) as the most frost-tolerant species within the Lolium-Festuca complex was used as a model for research aimed at identifying the cellular components involved in the cold acclimation (CA) of forage grasses. The work presented here also comprises the first comprehensive proteomic research on CA in a group of monocotyledonous species which are able to withstand winter conditions. Individual F. pratensis plants with contrasting levels of frost tolerance, high frost tolerant (HFT) and low frost tolerant (LFT) plants, were selected for comparative proteomic research. The work focused on the analysis of leaf protein accumulation before and after 2, 8, and 26 h, and 3, 5, 7, 14, and 21 d of CA, using high-throughput two-dimensional electrophoresis, and on the identification of proteins which were accumulated differentially between the selected plants by the application of mass spectrometry. The analyses of approximately 800 protein profiles revealed a total of 41 (5.1%) proteins that showed a minimum of a 1.5-fold difference in abundance, at a minimum of one time point of CA for HFT and LFT genotypes. It was shown that significant differences in profiles of protein accumulation between the analysed plants appeared relatively early during cold acclimation, most often after 26 h (on the 2nd day) of CA and one-half of the differentially accumulated proteins were all parts of the photosynthetic apparatus. Several proteins identified here have been reported to be differentially accumulated during cold conditions for the first time in this paper. The functions of the selected proteins in plant cells and their probable influence on the level of frost tolerance in F. pratensis, are discussed. PMID- 19553369 TI - Dormancy-associated MADS genes from the EVG locus of peach [Prunus persica (L.) Batsch] have distinct seasonal and photoperiodic expression patterns. AB - Mapping and sequencing of the non-dormant evg mutant in peach [Prunus persica (L.) Batsch] identified six tandem-arrayed DAM (dormancy-associated MADS-box) genes as candidates for regulating growth cessation and terminal bud formation. To narrow the list of candidate genes, an attempt was made to associate bud phenology with the seasonal and environmental patterns of expression of the candidates in wild-type trees. The expression of the six peach DAM genes at the EVG locus of peach was characterized throughout an annual growing cycle in the field, and under controlled conditions in response to a long day-short day photoperiod transition. DAM1, 2, 4, 5, and 6 were responsive to a reduction in photoperiod in controlled conditions and the direction of response correlated with the seasonal timing of expression in field-grown trees. DAM3 did not respond to photoperiod and may be regulated by chilling temperatures. The DAM genes in peach appear to have at least four distinct patterns of expression. DAM1, 2, and 4 are temporally associated with seasonal elongation cessation and bud formation and are the most likely candidates for control of the evg phenotype. PMID- 19553370 TI - Remobilization of leaf S compounds and senescence in response to restricted sulphate supply during the vegetative stage of oilseed rape are affected by mineral N availability. AB - The impact of sulphur limitation on the remobilization of endogenous S compounds during the rosette stage of oilseed rape, and the interactions with N availability on these processes, were examined using a long-term (34)SO(4)(2-) labelling method combined with a study of leaf senescence progression (using SAG12/Cab as a molecular indicator) and gene expression of the transporters, BnSultr4;1 and BnSultr4;2, involved in vacuolar sulphate efflux. After 51 d on hydroponic culture at 0.3 mM (34)SO(4)(2-) (1 atom% excess), the labelling was stopped and plants were subject for 28 d to High S-High N (HS-HN, control), Low S High N (LS-HN) or Low S-Low N (LS-LN) conditions. Compared with the control, LS HN plants showed delayed leaf senescence and, whilst the shoot growth and the foliar soluble protein amounts were not affected, S, (34)S, and SO(4)(2-) amounts in the old leaves declined rapidly and were associated with the up-regulation of BnSultr4;1. In LS-LN plants, shoot growth was reduced, leaf senescence was accelerated, and the rapid S mobilization in old leaves was accompanied by decreased (34)S and SO(4)(2-), higher protein mobilization, and up-regulation of BnSultr4;2, but without any change of expression of BnSultr4;1. The data suggest that to sustain the S demand for growth under S restriction (i) vacuolar SO(4)(2 ) is specifically remobilized in LS-HN conditions without any acceleration of leaf senescence, (ii) SO(4)(2-) mobilization is related to an up-regulation of BnSultr4;1 and/or BnSultr4;2 expression, and (iii) the relationship between sulphate mobilization and up-regulation of expression of BnSultr4 genes is specifically dependent on the N availability. PMID- 19553371 TI - Identification of genomic regions determining the phenological development leading to floral transition in wheat (Triticum aestivum L.). AB - Autumn-seeded winter cereals acquire tolerance to freezing temperatures and become vernalized by exposure to low temperature (LT). The level of accumulated LT tolerance depends on the cold acclimation rate and factors controlling timing of floral transition at the shoot apical meristem. In this study, genomic loci controlling the floral transition time were mapped in a winter wheat (T. aestivum L.) doubled haploid (DH) mapping population segregating for LT tolerance and rate of phenological development. The final leaf number (FLN), days to FLN, and days to anthesis were determined for 142 DH lines grown with and without vernalization in controlled environments. Analysis of trait data by composite interval mapping (CIM) identified 11 genomic regions that carried quantitative trait loci (QTLs) for the developmental traits studied. CIM analysis showed that the time for floral transition in both vernalized and non-vernalized plants was controlled by common QTL regions on chromosomes 1B, 2A, 2B, 6A and 7A. A QTL identified on chromosome 4A influenced floral transition time only in vernalized plants. Alleles of the LT-tolerant parent, Norstar, delayed floral transition at all QTLs except at the 2A locus. Some of the QTL alleles delaying floral transition also increased the length of vegetative growth and delayed flowering time. The genes underlying the QTLs identified in this study encode factors involved in regional adaptation of cold hardy winter wheat. PMID- 19553374 TI - Fulminant polyarteritis nodosa associated with acute myeloid leukaemia resulted in bilateral lower leg amputation. PMID- 19553372 TI - Tobacco plastid transformation using the feedback-insensitive anthranilate synthase [alpha]-subunit of tobacco (ASA2) as a new selectable marker. AB - Genetic engineering of chloroplasts normally requires the stable introduction of bacterial derived antibiotic or herbicide-resistance genes as selective markers. Ecological and health concerns have been raised due to the presence of such genes within the environment or the food supply. One way to overcome this issue is the use of plant genes able to confer a metabolic or developmental advantage to the transformed cells manipulating the plant's biosynthetic pathways. We explored the feasibility of using, for plastid transformation, the selection system based on the feedback-insensitive anthranilate synthase (AS) alpha-subunit gene of tobacco (ASA2) as a new selective marker and the indole analogue 4-methylindole (4MI) or the tryptophan analogue 7-methyl-DL-tryptophan (7MT) as the selection agents. An expression cassette containing Prrn-ASA2 was effectively integrated into the region between accD and ycf4 of the tobacco plastome by the biolistic process. Plastid transgenic plants were obtained on medium supplemented with 300 microM 7MT or 4MI. Transplastomic plants showed normal phenotype and fertility and the resistance to the selection agents 7MT and 4MI was transmitted maternally. The plastid transformed lines also exhibited a higher level of AS enzyme activity that was less sensitive to Trp-feedback inhibition and, consequently, increased free Trp levels in leaves about 7-fold. PMID- 19553375 TI - Bone mineral density in the hand as a predictor for mortality in patients with rheumatoid arthritis. AB - OBJECTIVES: BMD in the hand, as evaluated by digital X-ray radiogrammetry (DXR), has been suggested to be a predictor for joint damage in RA. A predictor for long term prognosis might also predict increased mortality in RA. The aim of the present study was to evaluate BMD in the hand as a predictor for all-cause mortality. METHODS: In 1978, 152 consecutive patients (78% women, mean disease duration: 14.2 years) were enrolled. X-rays of the hands at inclusion were available in 108 patients. Reasons for not evaluating DXR in 24 patients were placement of joint prostheses or severe malalignment. BMD was evaluated by DXR on the same digitized hand X-rays used for scoring radiographic joint damage. Measures of disease activity and damage were used to predict mortality by Cox regression models. RESULTS: From February 1978 through March 2008, 62 of the 82 patients died, corresponding to a standardized mortality ratio of 2.92 (95% CI 2.19, 3.65) for both sexes combined. In age- and sex-adjusted proportional hazards models, BMD [hazard ratio (HR) = 0.58/1 s.d.; 95% CI 0.37, 0.91], Steinbrocker functional class 3-4 (HR = 4.74/1 step; 95% CI 1.93, 11.64), the physician's global assessment (HR = 1.38/1 s.d.; 95% CI 1.03, 1.84) and ESR (HR = 1.92/1 s.d.; 95% CI 1.42, 2.58) were significant predictors of mortality, but RF, disease duration, Larsen index, Ritchie articular index and the patient's global assessment were not. CONCLUSION: Low DXR-BMD predicted overall mortality in age- and sex-adjusted analyses, which further supports it as a valid measurement of disease activity or damage and as having prognostic value. PMID- 19553373 TI - A dynamic interface for capsaicinoid systems biology. AB - Capsaicinoids are the pungent alkaloids that give hot peppers (Capsicum spp.) their spiciness. While capsaicinoids are relatively simple molecules, much is unknown about their biosynthesis, which spans diverse metabolisms of essential amino acids, phenylpropanoids, benzenoids, and fatty acids. Pepper is not a model organism, but it has access to the resources developed in model plants through comparative approaches. To aid research in this system, we have implemented a comprehensive model of capsaicinoid biosynthesis and made it publicly available within the SolCyc database at the SOL Genomics Network (http://www.sgn.cornell.edu). As a preliminary test of this model, and to build its value as a resource, targeted transcripts were cloned as candidates for nearly all of the structural genes for capsaicinoid biosynthesis. In support of the role of these transcripts in capsaicinoid biosynthesis beyond correct spatial and temporal expression, their predicted subcellular localizations were compared against the biosynthetic model and experimentally determined compartmentalization in Arabidopsis (Arabidopsis thaliana). To enable their use in a positional candidate gene approach in the Solanaceae, these genes were genetically mapped in pepper. These data were integrated into the SOL Genomics Network, a clade oriented database that incorporates community annotation of genes, enzymes, phenotypes, mutants, and genomic loci. Here, we describe the creation and integration of these resources as a holistic and dynamic model of the characteristic specialized metabolism of pepper. PMID- 19553376 TI - Health-related quality of life, employment and disability in patients with Sjogren's syndrome. AB - OBJECTIVE: To compare health-related quality of life (HR-QOL), employment and disability of primary and secondary SS (pSS and sSS, respectively) patients with the general Dutch population. METHODS: HR-QOL, employment and disability were assessed in SS patients regularly attending the University Medical Center Groningen (n = 235). HR-QOL, employment and disability were evaluated with the Short Form-36 questionnaire (SF-36) and an employment and disability questionnaire. Results were compared with Dutch population data (matched for sex and age). Demographical and clinical data associated with HR-QOL, employment and disability were assessed. RESULTS: Response rate was 83%. SS patients scored lower on HR-QOL than the general Dutch population. sSS patients scored lower on physical functioning, bodily pain and general health than pSS patients. Predictors for reduced HR-QOL were fatigue, tendomyalgia, articular involvement, use of artificial saliva, use of anti-depressants, comorbidity, male sex and eligibility for disability compensation (DC). Employment was lower and DC rates were higher in SS patients compared with the Dutch population. CONCLUSION: SS has a large impact on HR-QOL, employment and disability. PMID- 19553377 TI - Usefulness of erythrocyte-bound C4d as a biomarker to predict disease activity in patients with systemic lupus erythematosus. AB - OBJECTIVE: SLE is an autoimmune disorder characterized by abnormal complement activation. Numerous new biomarkers have recently been used to diagnose or monitor disease activity in patients with SLE. We checked the levels of erythrocyte-bound C4d (E-C4d), an activation-derived fragment of C4 that is deposited on the erythrocytes, under different conditions of SLE in order to correlate these levels with disease activity. METHODS: We conducted a cross sectional investigation of three groups of patients: (i) 63 patients with SLE; (ii) 43 patients with other diseases; and (iii) 26 healthy controls. Erythrocytes were analysed by flow cytometry to determine the levels of E-C4d. RESULTS: We found a significant elevation in the mean levels of E-C4d in SLE patients compared with patients with other diseases or healthy controls. In SLE patients, the levels of E-C4d were correlated with the SLEDAI and inversely correlated with serum C3/C4 levels. In the subgroup of SLE patients with haemolytic anaemia (HA), a significantly higher level of E-C4d was observed than that in SLE patients without HA. However, in SLE patients with HA, there was no correlation between the levels of E-C4d and other markers of disease activity, including SLEDAI and levels of anti-dsDNA, C3 and C4. CONCLUSION: E-C4d levels are useful diagnostic markers for SLE and can serve as biomarkers of disease activity in patients with SLE. However, E-C4d is of limited value in monitoring disease activity in SLE patients with HA. PMID- 19553378 TI - Making context memories independent of the hippocampus. AB - We present evidence that certain learning parameters can make a memory, even a very recent one, become independent of the hippocampus. We confirm earlier findings that damage to the hippocampus causes severe retrograde amnesia for context memories, but we show that repeated learning sessions create a context memory that is not vulnerable to the damage. The findings demonstrate that memories normally dependent on the hippocampus are incrementally strengthened in other memory networks with additional learning. The latter provides a new account for patterns of hippocampal retrograde amnesia and how memories may become independent of the hippocampus. PMID- 19553379 TI - DHPG activation of group 1 mGluRs in BLA enhances fear conditioning. AB - Group 1 metabotropic glutamate receptors are known to play an important role in both synaptic plasticity and memory. We show that activating these receptors prior to fear conditioning by infusing the group 1 mGluR agonist, (R.S.)-3,5 dihydroxyphenylglycine (DHPG), into the basolateral region of the amygdala (BLA) of adult Sprague-Dawley rats enhances freezing normally supported by a weak footshock. This effect of DHPG was blocked when it was co-infused with either the general group 1 mGluR1 antagonist, (R,S)-1-aminoindan-1,5 dicarboxylic acid (AIDA), or with the selective mGluR5 antagonist, 2-methyl-6-(phenylethynyl) pyridine (MPEP). These results support previous findings by Rodrigues and colleagues that mGluR5s in the lateral region of the amygdala make an import contribution to fear conditioning. More importantly, they support the general ideas embedded in the concept of metaplasticity, as per Abraham, and the synaptic tagging hypothesis per Frey and Morris-that the processes that specify the content of experience can be experimentally separated from those needed to acquire the memory. PMID- 19553380 TI - Side-specific reward memories in honeybees. AB - We report a hitherto unknown form of side-specific learning in honeybees. We trained bees individually by coupling gustatory and mechanical stimulation of each antenna with either increasing or decreasing volumes of sucrose solution offered to the animal's proboscis along successive learning trials. Next, we examined their proboscis extension response (PER) after stimulation of each antenna 1, 2, 3, and 24 h after training. The bees extended their proboscises earlier after stimulation of the antenna that had been coupled with increasing volumes than after stimulation of the antenna that had been coupled with decreasing volumes, thereby revealing short- and long-term side differences in the bees' PE reaction time. The bees' reaction time correlated well with the reaction time of the muscles M17. Long-term side differences in reaction time were prevented by repetitive antennal stimulation. Mechanosensory input was indispensable and sufficient for revealing side differences in reaction time. Such differences were specific to the gustatory input that the bees experienced during training. Our results show that side differences in the bees' PE reaction time depend upon the activation of side-specific reward memories. These memories are formed via the combined effect of a specific property of reward, i.e., that its magnitude increases or decreases over time, and side information seemingly relying on mechanosensory input. We present a learning procedure suitable to study reward learning in honeybees, which includes precise behavioral measures, physiological correlates of behavior, and within-animal controls. This procedure will prove fruitful in pharmacological and electrophysiological analyses of the neural substrates underlying reward memories in honeybees. PMID- 19553381 TI - Lateralized spatial and object memory encoding in entorhinal and perirhinal cortices. AB - The perirhinal and entorhinal cortices are critical components of the medial temporal lobe (MTL) declarative memory system. Study of their specific functions using blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging (fMRI), however, has suffered from severe magnetic susceptibility signal dropout resulting in poor temporal signal-to-noise (tSNR) and thus weak BOLD signal detectability. We have demonstrated that higher spatial resolution in the z-plane leads to improved BOLD fMRI signal quality in the anterior medial temporal lobes when using a 16-element surface coil array at 3 T (Tesla). Using this technique, the present study investigated the roles of the anterior medial temporal lobe, particularly the entorhinal and perirhinal cortices, in both object and spatial memory. Participants viewed a series of fractal images and were instructed to encode either the object's identity or location. Object and spatial recognition memory were tested after 18-sec delays. Both the perirhinal and entorhinal cortices were active during the object and spatial encoding tasks. In both regions, object encoding was biased to the left hemisphere, whereas spatial encoding was biased to the right. A similar hemispheric bias was evident for recognition memory. Recent animal studies suggest functional dissociations among regions of the entorhinal cortex for spatial vs. object processing. Our findings suggest that this process-specific distinction may be expressed in the human brain as a hemispheric division of labor. PMID- 19553382 TI - New hippocampal neurons are not obligatory for memory formation; cyclin D2 knockout mice with no adult brain neurogenesis show learning. AB - The role of adult brain neurogenesis (generating new neurons) in learning and memory appears to be quite firmly established in spite of some criticism and lack of understanding of what the new neurons serve the brain for. Also, the few experiments showing that blocking adult neurogenesis causes learning deficits used irradiation and various drugs known for their side effects and the results obtained vary greatly. We used a novel approach, cyclin D2 knockout mice (D2 KO mice), specifically lacking adult brain neurogenesis to verify its importance in learning and memory. D2 KO mice and their wild-type siblings were tested in several behavioral paradigms, including those in which the role of adult neurogenesis has been postulated. D2 KO mice showed no impairment in sensorimotor tests, with only sensory impairment in an olfaction-dependent task. However, D2 KO mice showed proper procedural learning as well as learning in context (including remote memory), cue, and trace fear conditioning, Morris water maze, novel object recognition test, and in a multifunctional behavioral system IntelliCages. D2 KO mice also demonstrated correct reversal learning. Our results suggest that adult brain neurogenesis is not obligatory in learning, including the kinds of learning where the role of adult neurogenesis has previously been strongly suggested. PMID- 19553383 TI - Odor-specific habituation arises from interaction of afferent synaptic adaptation and intrinsic synaptic potentiation in olfactory cortex. AB - Segmentation of target odorants from background odorants is a fundamental computational requirement for the olfactory system and is thought to be behaviorally mediated by olfactory habituation memory. Data from our laboratory have shown that odor-specific adaptation in piriform neurons, mediated at least partially by synaptic adaptation between the olfactory bulb outputs and piriform cortex pyramidal cells, is highly odor specific, while that observed at the synaptic level is specific only to certain odor features. Behavioral data show that odor habituation memory at short time constants corresponding to synaptic adaptation is also highly odor specific and is blocked by the same pharmacological agents as synaptic adaptation. Using previously developed computational models of the olfactory system we show here how synaptic adaptation and potentiation interact to create the observed specificity of response adaptation. The model analyzes the mechanisms underlying the odor specificity of habituation, the dependence on functioning cholinergic modulation, and makes predictions about connectivity to and within the piriform neural network. Predictions made by the model for the role of cholinergic modulation are supported by behavioral results. PMID- 19553384 TI - Generalization of conditioned fear along a dimension of increasing fear intensity. AB - The present study investigated the extent to which fear generalization in humans is determined by the amount of fear intensity in nonconditioned stimuli relative to a perceptually similar conditioned stimulus. Stimuli consisted of graded emotionally expressive faces of the same identity morphed between neutral and fearful endpoints. Two experimental groups underwent discriminative fear conditioning between a face stimulus of 55% fear intensity (conditioned stimulus, CS+), reinforced with an electric shock, and a second stimulus that was unreinforced (CS-). In Experiment 1 the CS- was a relatively neutral face stimulus, while in Experiment 2 the CS- was the most fear-intense stimulus. Before and following fear conditioning, skin conductance responses (SCR) were recorded to different morph values along the neutral-to-fear dimension. Both experimental groups showed gradients of generalization following fear conditioning that increased with the fear intensity of the stimulus. In Experiment 1 a peak shift in SCRs extended to the most fear-intense stimulus. In contrast, generalization to the most fear-intense stimulus was reduced in Experiment 2, suggesting that discriminative fear learning procedures can attenuate fear generalization. Together, the findings indicate that fear generalization is broadly tuned and sensitive to the amount of fear intensity in nonconditioned stimuli, but that fear generalization can come under stimulus control. These results reveal a novel form of fear generalization in humans that is not merely based on physical similarity to a conditioned exemplar, and may have implications for understanding generalization processes in anxiety disorders characterized by heightened sensitivity to nonthreatening stimuli. PMID- 19553385 TI - Feldenkrais method balance classes improve balance in older adults: a controlled trial. AB - The objective of this study was to investigate the effects of Feldenkrais Method balance classes on balance and mobility in older adults. This was a prospective non-randomized controlled study with pre/post measures. The setting for this study was the general community. A convenience sample of 26 community-dwelling older adults (median age 75 years) attending Feldenkrais Method balance classes formed the Intervention group. Thirty-seven volunteers were recruited for the Control group (median age 76.5 years). A series of Feldenkrais Method balance classes (the 33312Getting Grounded Gracefully33313 series), two classes per week for 10 weeks, were conducted. Main outcome measures were Activities-Specific Balance Confidence (ABC) questionnaire, Four Square Step Test (FSST), self selected gait speed (using GAITRite instrumented gait mat). At re-testing, the Intervention group showed significant improvement on all of the measures (ABC, P = .016, FSST, P = .001, gait speed, P < .001). The Control group improved significantly on one measure (FSST, P < .001). Compared to the Control group, the Intervention group made a significant improvement in their ABC score (P = .005), gait speed (P = .017) and FSST time (P = .022). These findings suggest that Feldenkrais Method balance classes may improve mobility and balance in older adults. PMID- 19553386 TI - Podophyllum hexandrum-Mediated Survival Protection and Restoration of Other Cellular Injuries in Lethally Irradiated Mice. AB - This study aims at the development of a safe and effective formulation to counter the effects of lethal irradiation. The sub-fraction (G-001M), prepared from Podophyllum hexandrum has rendered high degree of survival (>90%) at a dose of 6 mg kg(-1) body weight (intramuscular) in lethally irradiated mice. Therapeutic dose of G-001M, at about 20 times lower concentration than its LD(100), has revealed a DRF of 1.62. Comet assay studies in peripheral blood leukocytes have reflected that, treatment of G-001M before irradiation has significantly reduced DNA tail length (P < .001) and DNA damage score (P < .001), as compared to radiation-only group. Spleen cell counts in irradiated animals had declined drastically at the very first day of exposure, and the fall continued till the 5th day (P < .001). In the treated irradiated groups, there was a steep reduction in the counts initially, but this phase did not prolong. More than 60% decline in thymocytes of irradiated group animals was registered at 5 h of irradiation when compared with controls, and the fall progressed further downwards with the similar pace till 5th day of exposure (P < .001). At later intervals, thymus was found fully regressed. In G-001M pre-treated irradiated groups also, thymocytes decreased till the 5th day but thereafter rejuvenated and within 30 days of treatment the values were close to normal. Current studies have explicitly indicated that, G-001M in very small doses has not only rendered high survivability in lethally irradiated mice, but also protected their cellular DNA, besides supporting fast replenishment of the immune system. PMID- 19553387 TI - Regional distribution of clomipramine and desmethylclomipramine in rat brain and peripheral organs on chronic clomipramine administration. AB - The tricyclic antidepressant (TCA) clomipramine has been widely used in psychiatry for over 40 years. More recently, its therapeutic potential as an antineoplastic drug has been identified. However, there are no prior data on regional distribution in the brain of clomipramine and its primary metabolite (desmethylclomipramine) after chronic oral administration. The aim of this study was to determine the concentrations of clomipramine and desmethylclomipramine in different rat-brain regions and to compare those with levels in plasma and peripheral organs after chronic oral treatment of Sprague Dawley rats (15 mg/kg) for 14 days. The levels of both parent TCA and metabolite were analysed by high performance liquid chromatography in six brain regions (cortex, hypothalamus, hippocampus, striatum, brainstem and cerebellum), five peripheral organs and in plasma. Our data show that the cerebral cortex had the highest concentration of clomipramine (2.9 microg/mg), with successively lower concentrations in the hypothalamus, striatum, cerebellum, hippocampus and brainstem. Of the peripheral organs, the lungs and liver, had the highest levels of clomipramine, while in the heart, only the metabolite was detected. The plasma concentration (0.17 microg/ml or 0.48 microM) was comparable to that in the hippocampus and cerebellum (approximately 0.20 microg/mg). The differential distribution of clomipramine in different brain regions and the regional variation in clomipramine to desmethylclomipramine ratios have implications for the use of clomipramine in psychiatry and neuro-oncology. PMID- 19553389 TI - Quantitative evaluation of the role of Epstein-Barr virus immediate-early protein BZLF1 in B-cell transformation. AB - The Epstein-Barr virus (EBV) immediate-early transactivator BZLF1 plays a key role in switching EBV infection from the latent to the lytic form by stimulating the expression cascade of lytic genes; it also regulates the expression of several cellular genes. Recently, we reported that BZLF1 is expressed in primary human B cells early after EBV infection. To investigate whether this BZLF1 expression early after infection plays a role in the EBV-induced growth transformation of primary B cells, we generated BZLF1-knockout EBV and quantitatively evaluated its transforming ability compared with that of wild-type EBV. We found that the 50% transforming dose of BZLF1-knockout EBV was quite similar to that of wild-type EBV. Established lymphoblastoid cell lines (LCLs) harbouring BZLF1-knockout EBV were indistinguishable from LCLs harbouring wild type EBV in their pattern of latent gene expression and in their growth in vitro. Furthermore, the copy numbers of EBV episomes were very similar in the LCLs harbouring BZLF1-knockout EBV and in those harbouring wild-type EBV. These data indicate that disrupting BZLF1 expression in the context of the EBV genome, and the resultant inability to enter lytic replication, have little impact on the growth of LCLs and the steady-state copy number of EBV episomes in established LCLs. PMID- 19553390 TI - Synergism between the components of the bipartite major immediate-early transcriptional enhancer of murine cytomegalovirus does not accelerate virus replication in cell culture and host tissues. AB - Major immediate-early (MIE) transcriptional enhancers of cytomegaloviruses are key regulators that are regarded as determinants of virus replicative fitness and pathogenicity. The MIE locus of murine cytomegalovirus (mCMV) shows bidirectional gene-pair architecture, with a bipartite enhancer flanked by divergent core promoters. Here, we have constructed recombinant viruses mCMV-DeltaEnh1 and mCMV DeltaEnh2 to study the impact of either enhancer component on bidirectional MIE gene transcription and on virus replication in cell culture and various host tissues that are relevant to CMV disease. The data revealed that the two unipartite enhancers can operate independently, but synergize in enhancing MIE gene expression early after infection. Kick-start transcription facilitated by the bipartite enhancer configuration, however, did not ultimately result in accelerated virus replication. We conclude that virus replication, once triggered, proceeds with a fixed speed and we propose that synergism between the components of the bipartite enhancer may rather increase the probability for transcription initiation. PMID- 19553388 TI - High-throughput sequence analysis of variants of human cytomegalovirus strains Towne and AD169. AB - The genomes of commonly used variants of human cytomegalovirus (HCMV) strains Towne and AD169 each contain a substantial mutation in which a region (U(L)/b') at the right end of the long unique region has been replaced by an inverted duplication of a region from the left end of the genome. Using high-throughput technology, we have sequenced HCMV strain Towne (ATCC VR-977) and confirmed the presence of two variants, one exhibiting the replacement in U(L)/b' and the other intact in this region. Both variants are mutated in genes RL13, UL1, UL40, UL130, US1 and US9. We have also sequenced a novel AD169 variant (varUC) that is intact in U(L)/b' except for a small deletion that affects genes UL144, UL142, UL141 and UL140. Like other AD169 variants, varUC is mutated in genes RL5A, RL13, UL36 and UL131A. A subpopulation of varUC contains an additional deletion affecting genes IRS1, US1 and US2. PMID- 19553391 TI - Cellular factor YY1 downregulates the human papillomavirus 16 E6/E7 promoter, P97, in vivo and in vitro from a negative element overlapping the transcription initiation site. AB - Cellular factors that bind to cis sequences in the human papillomavirus 16 (HPV 16) upstream regulatory region (URR) positively and negatively regulate the viral E6 and E7 oncogene promoter, P97. DNase I footprinting has revealed the binding of cellular proteins to two previously undetected cis elements overlapping and 3' of the transcription-initiation site of the P97 promoter. Mutations within homologous motifs found in both of these cis elements abolished their negative function in vivo and the binding of the same cellular complex in vitro. This factor was identified as YY1 by complex mobility and binding specificity in comparison with vaccinia virus-expressed, purified recombinant YY1 protein and by antigenic reactivity with YY1 antisera. Cis mutations in the 'initiator' YY1 site activated the P97 promoter in vivo and in vitro. P97 was also activated threefold in vitro by depletion of endogenous YY1 with wild-type, but not mutant, YY1 oligonucleotides from the IgH kappa E3' enhancer. Furthermore, increasing concentrations of exogenous, purified recombinant YY1 repressed wild-type P97 transcript levels by up to threefold, but did not influence the P97 promoter mutated in the 'initiator' YY1 site. Thus, the promoter-proximal YY1 site was not necessary for correct transcription initiation at the P97 promoter, but was found to be required for downregulation of P97 transcription in vivo and in vitro. In contrast to other viral and cellular promoters, where YY1 is thought to function as a positive transcription-'initiator' factor, HPV-16 P97 transcription is downregulated by YY1 from a critical motif overlapping the transcription start site. PMID- 19553392 TI - New insights into processing of bovine viral diarrhea virus glycoproteins E(rns) and E1. AB - Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus within the family Flaviviridae. Its single-stranded RNA encodes a polyprotein that is cleaved co- and post-translationally by viral and cellular proteases. However, the cleavage between the envelope proteins E(rns) and E1 is still unexplained. In this study, an E(rns)-E1 protein could be identified and characterized with a new E1-specific antiserum. With bicistronic constructs bearing a deletion in the E(rns)-encoding region and expressing E(rns) or the E(rns)-E1 protein, it could be shown that this protein is not essential for virus replication. Furthermore, two putative cleavage sites were mutated in eukaryotic expression plasmids, as well as in full-length cDNA constructs. The mutation of position P3 of a potential signal peptide peptidase site abolished cleavage completely and no infectious virus progeny could be observed, indicating that cleavage of the E(rns)-E1 protein is indispensable for virus growth. PMID- 19553393 TI - Inhibition of coxsackievirus B3 and related enteroviruses by antiviral short interfering RNA pools produced using phi6 RNA-dependent RNA polymerase. AB - Coxsackievirus B3 (CBV3) is a member of the human enterovirus B species and a common human pathogen. Even though much is known about the enteroviral life cycle, no specific drugs are available to treat enterovirus infections. RNA interference (RNAi) has evolved to be an important tool for antiviral experimental therapies and gene function studies. We describe here a novel approach for RNAi against CBVs by using a short interfering (siRNA) pool covering 3.5 kb of CBV3 genomic sequence. The RNA-dependent RNA polymerase (RdRP) of bacteriophage phi6 was used to synthesize long double-stranded RNA (dsRNA) from a cloned region (nt 3837-7399) of the CBV3 genome. The dsRNA was cleaved using Dicer, purified and introduced to cells by transfection. The siRNA pool synthesized using the phi6 RdRP (phi6-siRNAs) was considerably more effective than single-site siRNAs. The phi6-siRNA pool also inhibited replication of other enterovirus B species, such as coxsackievirus B4 and coxsackievirus A9. PMID- 19553394 TI - Novel cytomegaloviruses in free-ranging and captive great apes: phylogenetic evidence for bidirectional horizontal transmission. AB - Wild great apes often suffer from diseases of unknown aetiology. This is among the causes of population declines. Because human cytomegalovirus (HCMV) is an important pathogen, especially in immunocompromised individuals, a search for cytomegaloviruses (CMVs) in deceased wild and captive chimpanzees, gorillas and orang-utans was performed. By using a degenerate PCR targeting four conserved genes (UL54-UL57), several distinct, previously unrecognized CMVs were found for each species. Sequences of up to 9 kb were determined for ten novel CMVs, located in the UL54-UL57 block. A phylogenetic tree was inferred for the ten novel CMVs, the previously characterized chimpanzee CMV, HCMV strains and Old World and New World monkey CMVs. The primate CMVs fell into four clades, containing New World monkey, Old World monkey, orang-utan and human CMVs, respectively, plus two clades that each contained both chimpanzee and gorilla isolates (termed CG1 and CG2). The tree loci of the first four clades mirrored those for their respective hosts in the primate tree, suggesting that these CMV lineages arose through cospeciation with host lineages. The CG1 and CG2 loci corresponded to those of the gorilla and chimpanzee hosts, respectively. This was interpreted as indicating that CG1 and CG2 represented CMV lineages that had arisen cospeciationally with the gorilla and chimpanzee lineages, respectively, with subsequent transfer within each clade between the host genera. Divergence dates were estimated and found to be consistent with overall cospeciational development of major primate CMV lineages. However, CMV transmission between chimpanzees and gorillas in both directions has also occurred. PMID- 19553395 TI - The QRS interval in patients treated with resynchronization therapy: which value? PMID- 19553397 TI - Heart failure in left-sided native valve infective endocarditis: characteristics, prognosis, and results of surgical treatment. AB - AIMS: Although congestive heart failure (CHF) represents the most common cause of death in native valve infective endocarditis (IE), recent data on the outcome of IE complicated by CHF are lacking. We aimed to analyse the characteristics and prognosis of patients with left-sided native valve IE complicated by CHF and to evaluate the impact of early surgery on 1 year outcome. METHODS AND RESULTS: Two hundred and fifty-nine consecutive patients with definite left-sided native valve IE according to the Duke criteria were included in this analysis. When compared with patients without CHF (n = 151), new heart murmur, high comorbidity index, aortic valve IE, and severe valve regurgitation were more frequently observed in CHF patients (n = 108, 41.6%). Mitral valve IE, embolic events and neurological events were less frequent in CHF patients. Congestive heart failure was independently predictive of in-hospital [OR 3.8 (1.7-9.0); P = 0.0013] and 1 year mortality [HR 1.8 (1.1-3.0); P = 0.007]. Early surgery was performed in 46% of CHF patients with a peri-operative mortality of 10%. In the CHF group, comorbidity index, Staphylococcus aureus IE, uncontrolled infection, and major neurological events were univariate predictors of 1 year mortality. Early surgery was independently associated with improved 1 year survival [HR 0.45 (0.22-0.93); P = 0.03]. CONCLUSION: Left-sided native valve IE complicated by CHF is more frequent in aortic IE and is associated with severe regurgitation. Congestive heart failure is an independent predictor of in-hospital and 1 year mortality. In CHF patients, early surgery is independently associated with reduced mortality and should be widely considered to improve outcome. PMID- 19553396 TI - Decreased beta-adrenergic responsiveness following hypertrophy occurs only in cardiomyocytes that also re-express beta-myosin heavy chain. AB - AIMS: Cardiac hypertrophy is associated with a reduction in the contractile response to beta-adrenergic stimulation, and with re-expression of foetal genes such as beta-myosin heavy chain (MHC). However, whether these two markers of pathology develop concordantly in the same individual cells or independently in different cells is not known. METHODS AND RESULTS: To answer this question, we examined the beta-adrenergic response of individual beta-MHC expressing and non expressing myocytes from hypertrophic hearts, using a previously generated mouse model (YFP/beta-MHC) in which a yellow fluorescent protein (YFP) is fused to the native beta-MHC protein allowing easy identification of beta-MHC expressing cells. Yellow fluorescent protein/beta-MHC mice were submitted to 4 weeks of transverse aortic constriction (TAC), and the contractile parameters of isolated individual myocytes in response to the beta-adrenergic agonist isoproterenol were assessed. Our results demonstrate that the decrease in isoproterenol-induced cell shortening that develops in TAC hearts occurs only in those hypertrophic myocytes that re-express beta-MHC. Hypertrophic myocytes that do not express beta-MHC have contractility indices indistinguishable from non-TAC controls. CONCLUSION: These data show that the reduction of beta-adrenergic response occurs only in subsets, rather than in all myocytes, and is coincident with re-expression of beta-MHC. PMID- 19553398 TI - A meta-analysis of the prognostic significance of atrial fibrillation in chronic heart failure. AB - AIMS: Atrial fibrillation (AF) is one of the commonest sustained arrhythmias in chronic heart failure (CHF), although the prognostic implications of the presence of AF in CHF remain controversial. We have therefore performed this meta-analysis to study the effects of the presence of AF on mortality in CHF patients. METHODS AND RESULTS: A systematic MEDLINE search for all randomized trials and observational studies in which the influence of AF on CHF mortality was investigated and meta-analysis of the mortality data was performed. A total of 16 studies were identified of which 7 were randomized trials and 9 were observational studies including 30,248 and 23,721 patients, respectively. An adjusted meta-analysis of the data revealed that the presence of AF is associated with an adverse effect on total mortality with an odds ratio (OR) of 1.40 [95% confidence interval (CI) 1.32-1.48, P < 0.0001] in randomized trials and an OR of 1.14 (95% CI 1.03-1.26, P < 0.05) in observational studies. This increase in mortality associated with the presence of AF was observed in subgroups of CHF patients with both preserved and impaired left ventricular (LV) systolic function. CONCLUSION: In conclusion, meta-analysis of 16 studies involving 53,969 patients suggests that the presence of AF is associated with an adverse prognosis in CHF irrespective of LV systolic function. PMID- 19553399 TI - Stroke work or systolic dP/dtmax to evaluate acute response to cardiac resynchronization therapy: are they interchangeable? AB - BACKGROUND: Cardiac resynchronization therapy (CRT) is characterized by a approximately 30% non-response. Invasive haemodynamic measurements are a traditional method to evaluate response to CRT. This study evaluates the correlation between acute changes in dP/dt(max) and Stroke Work (SW) during CRT. METHODS: Thirty-four CRT candidates were haemodynamically evaluated by pressure volume loop analysis during biventricular pacing. RESULTS: Mean dP/dt(max) and SW at baseline were 854 +/- 198 and 5186 +/- 2349, and displayed an increase during pacing of 106 +/- 117 mmHg/s (13% +/- 14%) and 1303 +/- 3039 mL/mmHg (30% +/- 52%), respectively. No correlation was found between the percentage change in dP/dt(max) and SW (R = 0.06, P = ns). When defining response an augmentation of 10% relative to baseline for both parameters, 16 patients demonstrated an ambiguous response. CONCLUSION: Although both parameters display an average increase during pacing, the change relative to baseline values of SW and dP/dt(max) is not related. PMID- 19553400 TI - A case of mucin-producing cholangiocarcinoma. PMID- 19553401 TI - Comparison of time trends in prostate cancer incidence (1973-2002) in Asia, from cancer incidence in five continents, Vols IV-IX. PMID- 19553402 TI - Finding sequence motifs in prokaryotic genomes--a brief practical guide for a microbiologist. AB - Finding significant nucleotide sequence motifs in prokaryotic genomes can be divided into three types of tasks: (1) supervised motif finding, where a sample of motif sequences is used to find other similar sequences in genomes; (2) unsupervised motif finding, which typically relates to the task of finding regulatory motifs and protein binding sites and (3) exploratory motif finding, which aims to identify potential functionally significant sequence motifs as those that are unusual in some statistical sense. This article provides a conceptual overview for each type of task, a brief description of basic algorithms used in their solution, and a review of selected relevant software available online. PMID- 19553403 TI - Propofol clearance and volume of distribution are increased in patients with major burns. AB - Propofol pharmacokinetics were examined in 17 adults with major burns during the hyperdynamic convalescent phase. Eighteen nonburned surgical patients served as controls. After a 2-mg/kg intravenous dose of propofol, blood samples were collected at multiple time points. Noncompartmental methods were used to calculate the pharmacokinetic parameters. The following indices were higher in burns than controls: propofol clearance (64+/-17 vs 29+/-4 mL/kg/min, P<.001) and volume of distribution (8+/-3 vs 4+/-1 L/kg, P<.001). The area under the curve was smaller in burns (556+/-152 vs 1193+/-183 microg/mL h, P<.001). The higher clearance of propofol in burned patients may imply that these patients require higher doses or infusion rates of propofol to attain a target plasma concentration or pharmacodynamic effect. PMID- 19553404 TI - Pharmacokinetics, safety, and tolerability of doripenem after 0.5-, 1-, and 4 hour infusions in healthy volunteers. AB - The pharmacokinetics, safety, and tolerability of doripenem in healthy subjects were evaluated in 2 studies. Study 1 was a double-blind, randomized, placebo controlled dose-escalation study in which doripenem was administered for 7 days by infusion over 30 minutes (500 mg) or 1 hour (1000 mg). Study 2 was an open label, randomized, 3-way crossover study in which each subject received a single dose of each of the following doripenem treatments on separate occasions: 500 mg infused over 1 hour, 500 mg infused over 4 hours, and 1000 mg infused over 4 hours. Doripenem exhibited linear pharmacokinetics with concordance between the studies for pharmacokinetic parameters. Doripenem did not accumulate with repeated dosing over 7 days. The area under the plasma concentration-time curve (AUC) for doripenem 500 mg infused over 1 hour versus 4 hours was bioequivalent, and the AUC and Cmax increased proportionally with dose for the 500- and 1000-mg doses administered over 4 hours. These results, along with the stability profile of doripenem, support its use as a prolonged infusion. All regimens of doripenem were safe and well tolerated. PMID- 19553406 TI - Methylphenidate has positive hypocholesterolemic and hypotriglyceridemic effects: new data. AB - Many psychotropic drugs may affect plasma lipids profile and their metabolism, with carbamazepine being the best known among them. Methylphenidate is a piperidine derivative structurally related to amphetamines and acts as a central nervous system stimulant. Its effect on lipid metabolism has not been investigated. The authors evaluated how methylphenidate affects the lipid profile in the plasma of patients diagnosed as having attention-deficit hyperactivity disorder (ADHD). All consecutive patients undergoing treatment for ADHD at the Adolescent Psychiatric Clinic (2003-2007) were enrolled. Blood samples for total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides, apolipoprotein A, apolipoprotein B, and lipoprotein (a) (Lp(a)) were collected before starting treatment and after 3 months of continuous treatment. Forty-two patients (22 men), median age 16, participated. The median total cholesterol count decreased by 9 mg/dL (P<.0002), LDL-C decreased by 5.0 mg/dL (P<.016), and triglycerides decreased by 8.0 mg/dL (P<.016). Changes in the levels of HDL-C, apolipoprotein A, and apolipoprotein B were nonsignificant, and Lp(a) levels decreased by 2.0 mg/dL (P<.0007). Methylphenidate improves the lipid profile by decreasing total cholesterol, triglycerides, LDL-C, and Lp(a). PMID- 19553405 TI - The absence of an interaction between warfarin and cranberry juice: a randomized, double-blind trial. AB - The question of potentiation of warfarin anticoagulation by cranberry juice (CJ) is a topic of biomedical importance. Anecdotal reports of CJ-warfarin interaction are largely unconfirmed in controlled studies. Thirty patients on stable warfarin anticoagulation (international normalized ratio [INR], 1.7-3.3) were randomized to receive 240 mL of CJ or 240 mL of placebo beverage, matched for color and taste, once daily for 2 weeks. The INR values and plasma levels of R- and S warfarin were measured during the 2-week period and a 1-week follow-up period. The CJ and placebo groups (n=14 and 16, respectively) did not differ significantly in mean plasma R- and S-warfarin concentrations. Eight patients (4 on CJ, 4 on placebo) developed minimally elevated INR (range, 3.38-4.52) during the treatment period. Mean INR differed significantly (P<.02) only on treatment day 12; at all other time points, the groups did not differ. Cranberry juice has no effect on plasma S- or R-warfarin plasma levels, excluding a pharmacokinetic interaction. A small though statistically significant pharmacodynamic enhancement of INR by CJ at a single time point is unlikely to be clinically important and may be a random change. Enhanced warfarin anticoagulation attributed to CJ in anecdotal reports may represent a chance temporal association. PMID- 19553407 TI - Response of vitiligo to narrowband ultraviolet B and oral antioxidants. AB - Narrowband ultraviolet B phototherapy (NB-UVB) is the most widely and effective therapeutic option in vitiligo. Antioxidant supplementation has also been reported to be useful. The purpose of this study was to determine the efficacy of oral antioxidants with NB-UVB in the treatment of vitiligo. Twenty-four patients with stable vitiligo were recruited and divided randomly into 2 groups. They were treated with NB-UVB plus oral vitamin E in group A and with NB-UVB only in group B. Improvement was recorded according to the extent of repigmentation in the existing lesions. Both plasma malondialdehyde (MDA; product of lipid peroxidation) and reduced glutathione (GSH) were measured before and after treatment. Twenty patients completed the study. Marked to excellent repigmentation was noted in 72.7% and 55.6% of the patients in group A and group B, respectively. Of the patients, 70% in group A and 85% in group B experienced mild erythema. After treatment, there was a significant reduction in plasma MDA in group A than in group B, but the increase in plasma GSH was not significant. In conclusion, oral vitamin E may represent a valuable adjuvant therapy, preventing lipid peroxidation in the cellular membrane of melanocytes and increasing the effectiveness of NB-UVB. PMID- 19553408 TI - A beginner's guide to probability. PMID- 19553409 TI - A spotter's guide to study designs. PMID- 19553410 TI - Oral prednisolone did not improve outcomes in preschool children with an attack of virus-induced wheezing. PMID- 19553411 TI - Review: pneumococcal vaccination does not prevent pneumonia, bacteraemia, bronchitis, or mortality. PMID- 19553412 TI - Review: insufficient evidence exists for oral nutritional supplements as aids for recovery in treated active tuberculosis. PMID- 19553413 TI - Review: inhaled corticosteroids do not reduce mortality but increase pneumonia in chronic obstructive pulmonary disease. PMID- 19553414 TI - Community-based multidisciplinary screening and intervention by pharmacists and nurses reduced BP in diabetes. PMID- 19553415 TI - Review: evidence on dressings for superficial burns is of poor quality. PMID- 19553416 TI - Review: low molecular weight heparin reduces risk of venous thromboembolism in adults with leg immobilisation. PMID- 19553417 TI - Review: central venous catheters treated with anti-infective agents prevent catheter-related bloodstream infections. PMID- 19553419 TI - Review: admission-avoidance hospital-at-home decreases mortality at 6 months but does not differ from inpatient care for readmission. PMID- 19553418 TI - Review: alternative-foam mattresses and some operating-table overlays reduce pressure ulcers more than standard surfaces. PMID- 19553420 TI - Home-based cardiac rehab was as effective as hospital-based rehab in improving cardiac risk factors. PMID- 19553421 TI - Hospital and home rehabilitation did not differ for functional competence in activities of daily living. PMID- 19553422 TI - Intensive nutritional supplementation improved functional outcome after stroke. PMID- 19553423 TI - Review: exercise programmes prevent falls in elderly people. PMID- 19553424 TI - Review: long-term annual conversion rate to dementia was 3.3% in elderly people with mild cognitive impairment. PMID- 19553425 TI - Vaginal douching increased risk of sexually transmitted infections in high-risk adolescents. PMID- 19553426 TI - A multicomponent intervention improved diabetes care in primary care practices. PMID- 19553427 TI - A clinical prediction rule based on preoperative factors predicted the development of delirium after cardiac surgery. PMID- 19553428 TI - Review: the STRATIFY prediction tool has limited accuracy for predicting falls in hospital and geriatric rehabilitation inpatients. PMID- 19553429 TI - People who were obese tried diets but felt they needed ongoing support to empower them to make lifestyle changes. PMID- 19553431 TI - Specialist care was important for helping patients with cancer to live positively with malignant fungating wounds. PMID- 19553430 TI - Patients with cancer believed that chemotherapy had to "hurt" or "cause side effects" to be effective. PMID- 19553432 TI - Patients with end-stage COPD did not ask for help because they felt normal and did not realise the situation could be improved. PMID- 19553433 TI - Families with loved ones on mechanical ventilation in the ICU found a way to face "living with dying". PMID- 19553434 TI - Mean-variance or prospect theory? The nature of value representations in the human brain. PMID- 19553435 TI - Functional magnetic resonance imaging-assessed brain responses during an executive task depend on interaction of sleep homeostasis, circadian phase, and PER3 genotype. AB - Cognition is regulated across the 24 h sleep-wake cycle by circadian rhythmicity and sleep homeostasis through unknown brain mechanisms. We investigated these mechanisms in a functional magnetic resonance imaging study of executive function using a working memory 3-back task during a normal sleep-wake cycle and during sleep loss. The study population was stratified according to homozygosity for a variable-number (4 or 5) tandem-repeat polymorphism in the coding region of the clock gene PERIOD3. This polymorphism confers vulnerability to sleep loss and circadian misalignment through its effects on sleep homeostasis. In the less vulnerable genotype, no changes were observed in brain responses during the normal-sleep wake cycle. During sleep loss, these individuals recruited supplemental anterior frontal, temporal and subcortical regions, while executive function was maintained. In contrast, in the vulnerable genotype, activation in a posterior prefrontal area was already reduced when comparing the evening to the morning during a normal sleep-wake cycle. Furthermore, in the morning after a night of sleep loss, widespread reductions in activation in prefrontal, temporal, parietal and occipital areas were observed in this genotype. These differences occurred in the absence of genotype-dependent differences in circadian phase. The data show that dynamic changes in brain responses to an executive task evolve across the sleep-wake and circadian cycles in a regionally specific manner that is determined by a polymorphism which affects sleep homeostasis. The findings support a model of individual differences in executive control, in which the allocation of prefrontal resources is constrained by sleep pressure and circadian phase. PMID- 19553436 TI - Amyloid reduction by amyloid-beta vaccination also reduces mouse tau pathology and protects from neuron loss in two mouse models of Alzheimer's disease. AB - Shown to lower amyloid deposits and improve cognition in APP transgenic mouse models, immunotherapy appears to be a promising approach for the treatment of Alzheimer's disease (AD). Due to limitations in available animal models, however, it has been unclear whether targeting amyloid is sufficient to reduce the other pathological hallmarks of AD-namely, accumulation of pathological, nonmutated tau and neuronal loss. We have now developed two transgenic mouse models (APPSw/NOS2( /-) and APPSwDI/NOS2(-/-)) that more closely model AD. These mice show amyloid pathology, hyperphosphorylated and aggregated normal mouse tau, significant neuron loss, and cognitive deficits. A beta(1-42) or KLH vaccinations were started in these animals at 12 months, when disease progression and cognitive decline are well underway, and continued for 4 months. Vaccinated APPSwDI/NOS2(-/ ) mice, which have predominantly vascular amyloid pathology, showed a 30% decrease in brain A beta and a 35-45% reduction in hyperphosphorylated tau. Neuron loss and cognitive deficits were partially reduced. In APPSw/NOS2(-/-) vaccinated mice, brain A beta was reduced by 65-85% and hyperphosphorylated tau by 50-60%. Furthermore, neurons were completely protected, and memory deficits were fully reversed. Microhemorrhage was observed in all vaccinated APPSw/NOS2(-/ ) mice and remains a significant adverse event associated with immunotherapy. Nevertheless, by providing evidence that reducing amyloid pathology also reduces nonmutant tau pathology and blocks neuron loss, these data support the development of amyloid-lowering therapies for disease-modifying treatment of AD. PMID- 19553437 TI - GABA-cAMP response element-binding protein signaling regulates maturation and survival of newly generated neurons in the adult hippocampus. AB - Survival and integration of new neurons in the hippocampal circuit are rate limiting steps in adult hippocampal neurogenesis. Neuronal network activity is a major regulator of these processes, yet little is known about the respective downstream signaling pathways. Here, we investigate the role of cAMP response element-binding protein (CREB) signaling in adult hippocampal neurogenesis. CREB is activated in new granule neurons during a distinct developmental period. Loss of CREB function in a cell-autonomous manner impairs dendritic development, decreases the expression of the neurogenic transcription factor NeuroD and of the neuronal microtubule-associated protein, doublecortin (DCX), and compromises the survival of newborn neurons. In addition, GABA-mediated excitation regulates CREB activation at early developmental stages. Importantly, developmental defects after loss of GABA-mediated excitation can be compensated by enhanced CREB signaling. These results indicate that CREB signaling is a central pathway in adult hippocampal neurogenesis, regulating the development and survival of new hippocampal neurons downstream of GABA-mediated excitation. PMID- 19553439 TI - Complexin-I is required for high-fidelity transmission at the endbulb of Held auditory synapse. AB - Complexins (CPXs I-IV) presumably act as regulators of the SNARE (soluble N ethylmaleimide-sensitive factor attachment protein receptor) complex, but their function in the intact mammalian nervous system is not well established. Here, we explored the role of CPXs in the mouse auditory system. Hearing was impaired in CPX I knock-out mice but normal in knock-out mice for CPXs II, III, IV, and III/IV as measured by auditory brainstem responses. Complexins were not detectable in cochlear hair cells but CPX I was expressed in spiral ganglion neurons (SGNs) that give rise to the auditory nerve. Ca(2+)-dependent exocytosis of inner hair cells and sound encoding by SGNs were unaffected in CPX I knock-out mice. In the absence of CPX I, the resting release probability in the endbulb of Held synapses of the auditory nerve fibers with bushy cells in the cochlear nucleus was reduced. As predicted by computational modeling, bushy cells had decreased spike rates at sound onset as well as longer and more variable first spike latencies explaining the abnormal auditory brainstem responses. In addition, we found synaptic transmission to outlast the stimulus at many endbulb of Held synapses in vitro and in vivo, suggesting impaired synchronization of release to stimulus offset. Although sound encoding in the cochlea proceeds in the absence of complexins, CPX I is required for faithful processing of sound onset and offset in the cochlear nucleus. PMID- 19553440 TI - Spatial pattern coding of sensory information by climbing fiber-evoked calcium signals in networks of neighboring cerebellar Purkinje cells. AB - Climbing fiber input produces complex spike synchrony across populations of cerebellar Purkinje cells oriented in the parasagittal axis. Elucidating the fine spatial structure of this synchrony is crucial for understanding its role in the encoding and processing of sensory information within the olivocerebellar cortical circuit. We investigated these issues using in vivo multineuron two photon calcium imaging in combination with information theoretic analysis. Spontaneous dendritic calcium transients linked to climbing fiber input were observed in multiple neighboring Purkinje cells. Spontaneous synchrony of calcium transients between individual Purkinje cells falls off over approximately 200 microm mediolaterally, consistent with the presence of cerebellar microzones organized by climbing fiber input. Synchrony was increased after administration of harmaline, consistent with an olivary origin. Periodic sensory stimulation also resulted in a transient increase of synchrony after stimulus onset. To examine how synchrony affects the neural population code provided by the spatial pattern of complex spikes, we analyzed its information content. We found that spatial patterns of calcium events from small ensembles of cells provided substantially more stimulus information (59% more for seven-cell ensembles) than available by counting events across the pool without taking into account spatial origin. Information theoretic analysis indicated that, rather than contributing significantly to sensory coding via stimulus dependence, correlational effects on sensory coding are dominated by redundancy attributable to the prevalent spontaneous synchrony. The olivocerebellar circuit thus uses a labeled line code to report sensory signals, leaving open a role for synchrony in flexible selection of signals for output to deep cerebellar nuclei. PMID- 19553441 TI - Visual salience affects performance in a working memory task. AB - Many studies of bottom-up visual attention have focused on identifying which features of a visual stimulus render it salient--i.e., make it "pop out" from its background--and on characterizing the extent to which salience predicts eye movements under certain task conditions. However, few studies have examined the relationship between salience and other cognitive functions, such as memory. We examined the impact of visual salience in an object-place working memory task, in which participants memorized the position of 3-5 distinct objects (icons) on a two-dimensional map. We found that their ability to recall an object's spatial location was positively correlated with the object's salience, as quantified using a previously published computational model (Itti et al., 1998). Moreover, the strength of this relationship increased with increasing task difficulty. The correlation between salience and error could not be explained by a biasing of overt attention in favor of more salient icons during memorization, since eye tracking data revealed no relationship between an icon's salience and fixation time. Our findings show that the influence of bottom-up attention extends beyond oculomotor behavior to include the encoding of information into memory. PMID- 19553438 TI - Detection of interaural time differences in the alligator. AB - The auditory systems of birds and mammals use timing information from each ear to detect interaural time difference (ITD). To determine whether the Jeffress-type algorithms that underlie sensitivity to ITD in birds are an evolutionarily stable strategy, we recorded from the auditory nuclei of crocodilians, who are the sister group to the birds. In alligators, precisely timed spikes in the first order nucleus magnocellularis (NM) encode the timing of sounds, and NM neurons project to neurons in the nucleus laminaris (NL) that detect interaural time differences. In vivo recordings from NL neurons show that the arrival time of phase-locked spikes differs between the ipsilateral and contralateral inputs. When this disparity is nullified by their best ITD, the neurons respond maximally. Thus NL neurons act as coincidence detectors. A biologically detailed model of NL with alligator parameters discriminated ITDs up to 1 kHz. The range of best ITDs represented in NL was much larger than in birds, however, and extended from 0 to 1000 micros contralateral, with a median ITD of 450 micros. Thus, crocodilians and birds employ similar algorithms for ITD detection, although crocodilians have larger heads. PMID- 19553442 TI - Information about complex fingertip parameters in individual human tactile afferent neurons. AB - Although information in tactile afferent neurons represented by firing rates has been studied extensively over nearly a century, recent studies suggest that precise spike timing might be more important than firing rates. Here, we used information theory to compare the information content in the discharges of 92 tactile afferents distributed over the entire terminal segment of the fingertip when it was contacted by surfaces with different curvatures and force directions representative of everyday manipulations. Estimates of the information content with regard to curvature and force direction based on the precise timing of spikes were at least 2.2 times and 1.6 times, respectively, larger than that of spike counts during a 125 ms period of force increase. Moreover, the information regarding force direction based on the timing of the very first elicited spike was comparable with that provided by spike counts and more than twice as large with respect to object shape. For all encoding schemes, afferents terminating close to the stimulation site tended to convey more information about surface curvature than more remote afferents that tended to convey more information about force direction. Finally, coding schemes based on spike timing and spike counts overall contributed mostly independent information. We conclude that information about tactile stimuli in timing of spikes in primary afferents, even if limited to the first spikes, surpasses that contained in firing rates and that these measures of afferents' responses might capture different aspects of the stimulus. PMID- 19553443 TI - Searching for targets within the spatial layout of visual short-term memory. AB - Recent studies have revealed that the internal representations that we construct from the environment and maintain in visual short-term memory (VSTM) to guide behavior are highly flexible and can be selectively modulated according to our task goals and expectations. In the current study, we conducted two experiments to compare and contrast neural mechanisms of selective attention related to searching for target items within perceptual versus VSTM representations. We used event-related potentials to investigate whether searching for relevant target items from within VSTM representations involves spatially specific biasing of neural activity in a manner analogous to that which occurs during visual search for target items in perceptual arrays. The results, replicated across the two experiments, revealed that selection of a target object within a search array maintained in VSTM proceeds through a similar mechanism as that in the perceptual domain. In line with previous results, N2pc potentials were obtained when targets were identified within a perceptual visual-search array. Interestingly, equivalent N2pcs, with similar time courses and scalp distributions, were also elicited when target items were identified within a VSTM representation. The findings reinforce the notion of highly flexible VSTM representations that can be modulated according to task goals and suggest a large degree of overlap in the spatially specific neural mechanisms of target selection across the perceptual and VSTM domains. PMID- 19553444 TI - Neuroligin 2 controls the maturation of GABAergic synapses and information processing in the retina. AB - In the present study, we investigated the role of Neuroligin 2 (NL2) in synaptic transmission and network function using the mouse retina as a model circuit. We show that NL2 is preferentially located at GABAergic rather than glycinergic or glutamatergic postsynapses. The absence of NL2 from the retina resulted in a severe reduction of GABA(A) receptor clustering, and in subtle alterations of the retinal circuitry. Light processing was impaired accordingly, and retinal ganglion cells, the output neurons of the retina, showed increased basal activity and altered coding of visual information. Together, our data indicate that NL2 is essential for the functional integrity of GABAergic signaling and as a consequence, for information processing in the retina. PMID- 19553446 TI - Ca2+ regulation of dynamin-independent endocytosis in cortical astrocytes. AB - Astrocytes release ATP and glutamate through vesicular exocytosis to mediate neuron-glial interactions. In contrast to exocytosis, the endocytic pathways in astroglial cells are poorly understood. Here, we identify a constitutive endocytic pathway in cultured astrocytes that is dependent on neither clathrin nor dynamin. This dynamin-independent endocytic pathway is regulated by Rab5, an early endosome protein. The endocytosed vesicles show fast transition from early endosomes to late endosomes and lysosomes within a few minutes. Interestingly, this clathrin- and dynamin-independent endocytosis in astrocytes is potently regulated by intracellular Ca(2+). ATP and glutamate greatly enhance the dynamin independent endocytosis through elevating the intracellular Ca(2+). In addition, amyloid-beta peptide (A beta) also enhances the dynamin-independent endocytosis by inducing Ca(2+) transients in astrocytes. These results demonstrate a novel endocytic pathway in glial cells that is dynamin independent but tightly regulated by intracellular Ca(2+). The regulation by ATP, glutamate, and A beta suggests an important role of the dynamin-independent endocytosis in both physiological and pathological conditions. PMID- 19553445 TI - Visualization of chemokine receptor activation in transgenic mice reveals peripheral activation of CCR2 receptors in states of neuropathic pain. AB - CCR2 chemokine receptor signaling has been implicated in the generation of diverse types of neuropathology, including neuropathic pain. For example, ccr2 knock-out mice are resistant to the establishment of neuropathic pain, and mice overexpressing its ligand, monocyte chemoattractant protein-1 (MCP1; also known as CCL2), show enhanced pain sensitivity. However, whether CCR2 receptor activation occurs in the central or peripheral nervous system in states of neuropathic pain has not been clear. We developed a novel method for visualizing CCR2 receptor activation in vivo by generating bitransgenic reporter mice in which the chemokine receptor CCR2 and its ligand MCP1 were labeled by the fluorescent proteins enhanced green fluorescent protein and monomeric red fluorescent protein-1, respectively. CCR2 receptor activation under conditions such as acute inflammation and experimental autoimmune encephalomyelitis could be faithfully visualized by using these mice. We examined the status of CCR2 receptor activation in a demyelination injury model of neuropathic pain and found that MCP1-induced CCR2 receptor activation mainly occurred in the peripheral nervous system, including the injured peripheral nerve and dorsal root ganglia. These data explain the rapid antinociceptive effects of peripherally administered CCR2 antagonists under these circumstances, suggesting that CCR2 antagonists may ameliorate pain by inhibiting CCR2 receptor activation in the periphery. The method developed here for visualizing CCR2 receptor activation in vivo may be extended to G-protein-coupled receptors (GPCRs) in general and will be valuable for studying intercellular GPCR-mediated communication in vivo. PMID- 19553447 TI - Phosphodiesterase 5 inhibition improves synaptic function, memory, and amyloid beta load in an Alzheimer's disease mouse model. AB - Memory loss, synaptic dysfunction, and accumulation of amyloid beta-peptides (A beta) are major hallmarks of Alzheimer's disease (AD). Downregulation of the nitric oxide/cGMP/cGMP-dependent protein kinase/c-AMP responsive element-binding protein (CREB) cascade has been linked to the synaptic deficits after A beta elevation. Here, we report that the phosphodiesterase 5 inhibitor (PDE5) sildenafil (Viagra), a molecule that enhances phosphorylation of CREB, a molecule involved in memory, through elevation of cGMP levels, is beneficial against the AD phenotype in a mouse model of amyloid deposition. We demonstrate that the inhibitor produces an immediate and long-lasting amelioration of synaptic function, CREB phosphorylation, and memory. This effect is also associated with a long-lasting reduction of A beta levels. Given that side effects of PDE5 inhibitors are widely known and do not preclude their administration to a senile population, these drugs have potential for the treatment of AD and other diseases associated with elevated A beta levels. PMID- 19553448 TI - A cholinergic-dependent role for the entorhinal cortex in trace fear conditioning. AB - Trace conditioning is considered a model of higher cognitive involvement in simple associative tasks. Studies of trace conditioning have shown that cortical areas and the hippocampal formation are required to associate events that occur at different times. However, the mechanisms that bridge the trace interval during the acquisition of trace conditioning remain unknown. In four experiments with fear conditioning in rats, we explored the involvement of the entorhinal cortex (EC) in the acquisition of fear under a trace-30 s protocol. We first determined that pretraining neurotoxic lesions of the EC selectively impaired trace-, but not delay-conditioned fear as evaluated by freezing behavior. A local cholinergic deafferentation of the EC using 192-IgG-saporin did not replicate this deficit, presumably because cholinergic interneurons were spared by the toxin. However, pretraining local blockade of EC muscarinic receptors with the M1 antagonist pirenzepine yielded a specific and dose-dependent deficit in trace-conditioned responses. The same microinjections performed after conditioning were without effect on trace fear responses. These effects of blocking M1 receptors are consistent with the notion that conditioned stimulus (CS)-elicited, acetylcholine dependent persistent activities in the EC are needed to maintain a representation of a tone CS across the trace interval during the acquisition of trace conditioning. This function of the EC is consistent with recent views of this region as a short-term stimulus buffer. PMID- 19553449 TI - GABAergic neurons of the medial septum lead the hippocampal network during theta activity. AB - Information processing in the hippocampus critically relies on its reciprocal interaction with the medial septum (MS). Synchronization of the septo-hippocampal system was demonstrated during both major hippocampal activity states, the regular theta rhythm and the large amplitude irregular activity. Previous experimental and modeling data suggest that the MS provides rhythmic drive to the hippocampus, and hippocampo-septal feedback synchronizes septal pacemaker units. However, this view has recently been questioned based on the possibility of intrahippocampal theta genesis. Previously, we identified putative pacemaker neurons expressing parvalbumin (PV) and/or the pacemaker hyperpolarization activated and cyclic nucleotide-gated nonselective cation channel (HCN) in the MS. In this study, by analyzing the temporal relationship of activity between the PV/HCN-containing medial septal neurons and hippocampal local field potential, we aimed to uncover whether the sequence of events during theta formation supports the classic view of septal drive or the challenging theory of hippocampal pacing of theta. Importantly, by implementing a circular statistical method, a temporal lead of these septal neurons over the hippocampus was observed on the course of theta synchronization. Moreover, the activity of putative hippocampal interneurons also preceded hippocampal local field theta, but by a shorter time period compared with PV/HCN-containing septal neurons. Using the concept of mutual information, the action potential series of PV/HCN-containing neurons shared higher amount of information with hippocampal field oscillation than PV/HCN-immunonegative cells. Thus, a pacemaker neuron population of the MS leads hippocampal activity, presumably via the synchronization of hippocampal interneurons. PMID- 19553451 TI - Novelty enhancements in memory are dependent on lateral prefrontal cortex. AB - Physiological evidence indicates that several brain regions, including the medial temporal lobes and prefrontal cortex (PFC), are involved in processing events that are novel or distinctive in their immediate context. However, behavioral studies that investigate whether these regions are critical for producing stimulus novelty advantages in memory are limited. For example, evidence from an animal lesion study indicated that the PFC is involved in stimulus novelty effects, but this has not been examined in humans. In the current study, we used a von Restorff novelty paradigm to test a large cohort of lateral PFC patients (n = 16). We found that patients with lateral PFC damage were impaired in recollection- and familiarity-based recognition, and they did not exhibit a normal memory advantage for novel compared with non-novel items. These results provide neuropsychological evidence supporting a key role for the lateral PFC in producing stimulus novelty advantages in memory. PMID- 19553450 TI - Nonmotor symptoms of Parkinson's disease revealed in an animal model with reduced monoamine storage capacity. AB - Parkinson's disease (PD) is a progressive neurodegenerative disorder that is characterized by the loss of dopamine neurons in the substantia nigra pars compacta, culminating in severe motor symptoms, including resting tremor, rigidity, bradykinesia, and postural instability. In addition to motor deficits, there are a variety of nonmotor symptoms associated with PD. These symptoms generally precede the onset of motor symptoms, sometimes by years, and include anosmia, problems with gastrointestinal motility, sleep disturbances, sympathetic denervation, anxiety, and depression. Previously, we have shown that mice with a 95% genetic reduction in vesicular monoamine transporter expression (VMAT2 deficient, VMAT2 LO) display progressive loss of striatal dopamine, L-DOPA responsive motor deficits, alpha-synuclein accumulation, and nigral dopaminergic cell loss. We hypothesized that since these animals exhibit deficits in other monoamine systems (norepinephrine and serotonin), which are known to regulate some of these behaviors, the VMAT2-deficient mice may display some of the nonmotor symptoms associated with PD. Here we report that the VMAT2-deficient mice demonstrate progressive deficits in olfactory discrimination, delayed gastric emptying, altered sleep latency, anxiety-like behavior, and age-dependent depressive behavior. These results suggest that the VMAT2-deficient mice may be a useful model of the nonmotor symptoms of PD. Furthermore, monoamine dysfunction may contribute to many of the nonmotor symptoms of PD, and interventions aimed at restoring monoamine function may be beneficial in treating the disease. PMID- 19553452 TI - Cholinergic stimulation enhances neural activity associated with encoding but reduces neural activity associated with retrieval in humans. AB - The cerebral cholinergic system is centrally involved in memory formation. Studies in rodents suggest that cholinergic stimulation may facilitate encoding of new information but may interfere with retrieval. We investigated the effect of cholinergic stimulation on encoding and retrieval of episodic memory in humans. We also tested whether the putative benefit of cholinergic stimulation on memory function depends on individual baseline performance. Since such effects were expected to be greatest in an older population resulting from an age-related degeneration of the cholinergic system, we recruited 22 healthy older subjects (51-68 years) for an event-related functional magnetic resonance imaging experiment. In two separate scanning sessions, subjects encoded and retrieved items and their spatial context under cholinergic stimulation or placebo with the acetylcholine-esterase inhibitor physostigmine or saline being administered intravenously in a double-blind cross-over design. Baseline performance was recorded at a separate occasion without scanning. Cholinergic stimulation enhanced neural activity for successful versus unsuccessful spatial context encoding in the right hippocampus but reduced activity for successful versus unsuccessful spatial context retrieval in the right amygdala. These data may bridge the gap between rodent and human studies by showing that also in man cholinergic stimulation enhances encoding but interferes with retrieval on a neural level. Furthermore, baseline performance negatively correlated with the effect of cholinergic stimulation. Thus, participants who were worse at baseline benefited more from cholinergic stimulation than those who had better baseline values, indicating that a cholinergic deficit contributes to the memory decline even in healthy older subjects. PMID- 19553454 TI - Glutamate transporter coupling to Na,K-ATPase. AB - Deactivation of glutamatergic signaling in the brain is mediated by glutamate uptake into glia and neurons by glutamate transporters. Glutamate transporters are sodium-dependent proteins that putatively rely indirectly on Na,K-ATPases to generate ion gradients that drive transmitter uptake. Based on anatomical colocalization, mutual sodium dependency, and the inhibitory effects of the Na,K ATPase inhibitor ouabain on glutamate transporter activity, we postulated that glutamate transporters are directly coupled to Na,K-ATPase and that Na,K-ATPase is an essential modulator of glutamate uptake. Na,K-ATPase was purified from rat cerebellum by tandem anion exchange and ouabain affinity chromatography, and the cohort of associated proteins was characterized by mass spectrometry. The alpha1 alpha 3 subunits of Na,K-ATPase were detected, as were the glutamate transporters GLAST and GLT-1, demonstrating that glutamate transporters copurify with Na,K ATPases. The link between glutamate transporters and Na,K-ATPase was further established by coimmunoprecipitation and colocalization. Analysis of the regulation of glutamate transporter and Na,K-ATPase activities was assessed using [(3)H]D-aspartate, [(3)H]L-glutamate, and rubidium-86 uptake into synaptosomes and cultured astrocytes. In synaptosomes, ouabain produced a dose-dependent inhibition of glutamate transporter and Na,K-ATPase activities, whereas in astrocytes, ouabain showed a bimodal effect whereby glutamate transporter activity was stimulated at 1 microm ouabain and inhibited at higher concentrations. The effects of protein kinase inhibitors on [(3)H]D-aspartate uptake indicated the selective involvement of Src kinases, which are probably a component of the Na,K-ATPase/glutamate transporter complex. These findings demonstrate that glutamate transporters and Na,K-ATPases are part of the same macromolecular complexes and operate as a functional unit to regulate glutamatergic neurotransmission. PMID- 19553453 TI - Focal adhesion kinase acts downstream of EphB receptors to maintain mature dendritic spines by regulating cofilin activity. AB - Dendritic spines are the postsynaptic sites of most excitatory synapses in the brain and are highly enriched in polymerized F-actin, which drives the formation and maintenance of mature dendritic spines and synapses. We propose that suppressing the activity of the actin-severing protein cofilin plays an important role in the stabilization of mature dendritic spines, and is accomplished through an EphB receptor-focal adhesion kinase (FAK) pathway. Our studies revealed that Cre-mediated knock-out of loxP-flanked fak prompted the reversion of mature dendritic spines to an immature filopodial-like phenotype in primary hippocampal cultures. The effects of FAK depletion on dendritic spine number, length, and morphology were rescued by the overexpression of the constitutively active FAK(Y397E), but not FAK(Y397F), indicating the significance of FAK activation by phosphorylation on tyrosine 397. Our studies demonstrate that FAK acts downstream of EphB receptors in hippocampal neurons and EphB2-FAK signaling controls the stability of mature dendritic spines by promoting cofilin phosphorylation, thereby inhibiting cofilin activity. While constitutively active nonphosphorylatable cofilin(S3A) induced an immature spine profile, phosphomimetic cofilin(S3D) restored mature spine morphology in neurons with disrupted EphB activity or lacking FAK. Further, we found that EphB-mediated regulation of cofilin activity at least partially depends on the activation of Rho-associated kinase (ROCK) and LIMK-1. These findings indicate that EphB2 mediated dendritic spine stabilization relies, in part, on the ability of FAK to activate the RhoA-ROCK-LIMK-1 pathway, which functions to suppress cofilin activity and inhibit cofilin-mediated dendritic spine remodeling. PMID- 19553456 TI - Functional significance of nonspatial information in monkey lateral intraparietal area. AB - Although the parietal cortex is traditionally associated with spatial perception and motor planning, recent evidence shows that neurons in the lateral intraparietal area (LIP) carry both spatial and nonspatial signals. The functional significance of the nonspatial information in the parietal cortex is not understood. To address this question, we tested the effect of unilateral reversible inactivation of LIP on three behavioral tasks known to evoke nonspatial responses. Each task included a spatial component (target selection in the hemifield contralateral or ipsilateral to the inactivation) and a nonspatial component, namely limb motor planning, the estimation of elapsed time, and reward based decisions. Although inactivation reliably impaired performance on all tasks, the deficits were spatially specific (restricted to contralateral target locations), and there were no effects on nonspatial aspects on performance. This suggests that modulatory nonspatial signals in LIP represent feedback about computations performed elsewhere rather than a primary role of LIP in these computations. PMID- 19553455 TI - Impact of serotonin 2C receptor null mutation on physiology and behavior associated with nigrostriatal dopamine pathway function. AB - The impact of serotonergic neurotransmission on brain dopaminergic pathways has substantial relevance to many neuropsychiatric disorders. A particularly prominent role has been ascribed to the inhibitory effects of serotonin 2C receptor (5-HT(2C)R) activation on physiology and behavior mediated by the mesolimbic dopaminergic pathway, particularly in the terminal region of the nucleus accumbens. The influence of this receptor subtype on functions mediated by the nigrostriatal dopaminergic pathway is less clear. Here we report that a null mutation eliminating expression of 5-HT(2C)Rs produces marked alterations in the activity and functional output of this pathway. 5-HT(2C)R mutant mice displayed increased activity of substantia nigra pars compacta (SNc) dopaminergic neurons, elevated baseline extracellular dopamine concentrations in the dorsal striatum (DSt), alterations in grooming behavior, and enhanced sensitivity to the stereotypic behavioral effects of d-amphetamine and GBR 12909. These psychostimulant responses occurred in the absence of phenotypic differences in drug-induced extracellular dopamine concentration, suggesting a phenotypic alteration in behavioral responses to released dopamine. This was further suggested by enhanced behavioral responses of mutant mice to the D(1) receptor agonist SKF 81297. Differences in DSt D(1) or D(2) receptor expression were not found, nor were differences in medium spiny neuron firing patterns or intrinsic membrane properties following dopamine stimulation. We conclude that 5-HT(2C)Rs regulate nigrostriatal dopaminergic activity and function both at SNc dopaminergic neurons and at a locus downstream of the DSt. PMID- 19553457 TI - Opioids depress cortical centers responsible for the volitional control of respiration. AB - Respiratory depression limits provision of safe opioid analgesia and is the main cause of death in drug addicts. Although opioids are known to inhibit brainstem respiratory activity, their effects on cortical areas that mediate respiration are less well understood. Here, functional magnetic resonance imaging was used to examine how brainstem and cortical activity related to a short breath hold is modulated by the opioid remifentanil. We hypothesized that remifentanil would differentially depress brain areas that mediate sensory-affective components of respiration over those that mediate volitional motor control. Quantitative measures of cerebral blood flow were used to control for hypercapnia-induced changes in blood oxygen level-dependent (BOLD) signal. Awareness of respiration, reflected by an urge-to-breathe score, was profoundly reduced with remifentanil. Urge to breathe was associated with activity in the bilateral insula, frontal operculum, and secondary somatosensory cortex. Localized remifentanil-induced decreases in breath hold-related activity were observed in the left anterior insula and operculum. We also observed remifentanil-induced decreases in the BOLD response to breath holding in the left dorsolateral prefrontal cortex, anterior cingulate, the cerebellum, and periaqueductal gray, brain areas that mediate task performance. Activity in areas mediating motor control (putamen, motor cortex) and sensory-motor integration (supramarginal gyrus) were unaffected by remifentanil. Breath hold-related activity was observed in the medulla. These findings highlight the importance of higher cortical centers in providing contextual awareness of respiration that leads to appropriate modulation of respiratory control. Opioids have profound effects on the cortical centers that control breathing, which potentiates their actions in the brainstem. PMID- 19553458 TI - Inosine alters gene expression and axonal projections in neurons contralateral to a cortical infarct and improves skilled use of the impaired limb. AB - Recovery after stroke and other types of brain injury is restricted in part by the limited ability of undamaged neurons to form compensatory connections. Inosine, a naturally occurring purine nucleoside, stimulates neurons to extend axons in culture and, in vivo, enhances the ability of undamaged neurons to form axon collaterals after brain damage. The molecular changes induced by inosine are unknown, as is the ability of inosine to restore complex functions associated with a specific cortical area. Using a unilateral injury model limited to the sensorimotor cortex, we show that inosine triples the number of corticospinal tract axons that project from the unaffected hemisphere and form synaptic bouton like structures in the denervated half of the spinal cord. These changes correlate with improved recovery in animals' ability to grasp and consume food pellets with the affected forepaw. Studies using laser-capture microdissection and microarray analysis show that inosine profoundly affects gene expression in corticospinal neurons contralateral to the injury. Inosine attenuates transcriptional changes caused by the stroke, while upregulating the expression of genes associated with axon growth and the complement cascade. Thus, inosine alters gene expression in neurons contralateral to a stroke, enhances the ability of these neurons to form connections on the denervated side of the spinal cord, and improves performance with the impaired limb. PMID- 19553459 TI - Subcellular dynamics of somatostatin receptor subtype 1 in the rat arcuate nucleus: receptor localization and synaptic connectivity vary in parallel with the ultradian rhythm of growth hormone secretion. AB - Growth hormone (GH) secretion in male rats exhibits a 3.3 h ultradian rhythm generated by the reciprocal interaction of GH-releasing hormone (GHRH) and somatostatin (SRIF). SRIF receptor subtypes sst(1) and sst(2) are highly expressed in GHRH neurons of the hypothalamic arcuate nucleus (ARC). We previously demonstrated an ultradian oscillation in binding of SRIF analogs to the ARC in relation to GH peaks and troughs. Here we tested the hypothesis that these ultradian changes in SRIF binding are due to differential plasma membrane targeting of sst(1) receptors in ARC neurons using immunocytochemistry and electron microscopy. We found that 87% of sst(1)-positive ARC neurons also synthesized GHRH. Subcellularly, 80% of sst(1) receptors were located intracellularly and 20% at the plasma membrane regardless of GH status. However, whereas 30% of the cell-surface sst(1) receptors were located perisynaptically or subsynaptically following exposure to high GH secretion, this fraction was increased to 42% following a GH trough period (p = 0.05). Furthermore, the relative abundance of symmetric and asymmetric synapses on sst(1)-positive dendrites also varied significantly, depending on the GH cycle, from approximately equal numbers following GH troughs to 70:30 in favor of symmetric, i.e., inhibitory, inputs after GH peaks (p < 0.02). These findings suggest that postsynaptic localization of sst(1) receptors and synaptic connectivity in the ARC undergo pronounced remodeling in parallel with the GH rhythm. Such synaptic plasticity may be an important mechanism by which sst(1) mediates SRIF's cyclical effects on ARC GHRH neurons to generate the ultradian rhythm of GH secretion. PMID- 19553460 TI - The formation of recent and remote memory is associated with time-dependent formation of dendritic spines in the hippocampus and anterior cingulate cortex. AB - Although hippocampal-cortical interactions are crucial for the formation of enduring declarative memories, synaptic events that govern long-term memory storage remain mostly unclear. We present evidence that neuronal structural changes, i.e., dendritic spine growth, develop sequentially in the hippocampus and anterior cingulate cortex (aCC) during the formation of recent and remote contextual fear memory. We found that mice placed in a conditioning chamber for one 7 min conditioning session and exposed to five footshocks (duration, 2 s; intensity, 0.7 mA; interstimulus interval, 60 s) delivered through the grid floor exhibited robust fear response when returned to the experimental context 24 h or 36 d after the conditioning. We then observed that their fear response at the recent, but not the remote, time point was associated with an increase in spine density on hippocampal neurons, whereas an inverse temporal pattern of spine density changes occurred on aCC neurons. At each time point, hippocampal or aCC structural alterations were achieved even in the absence of recent or remote memory tests, thus suggesting that they were not driven by retrieval processes. Furthermore, ibotenic lesions of the hippocampus impaired remote memory and prevented dendritic spine growth on aCC neurons when they were performed immediately after the conditioning, whereas they were ineffective when performed 24 d later. These findings reveal that gradual structural changes modifying connectivity in hippocampal-cortical networks underlie the formation and expression of remote memory, and that the hippocampus plays a crucial but time limited role in driving structural plasticity in the cortex. PMID- 19553461 TI - Hyperdopaminergia and NMDA receptor hypofunction disrupt neural phase signaling. AB - Neural phase signaling has gained attention as a putative coding mechanism through which the brain binds the activity of neurons across distributed brain areas to generate thoughts, percepts, and behaviors. Neural phase signaling has been shown to play a role in various cognitive processes, and it has been suggested that altered phase signaling may play a role in mediating the cognitive deficits observed across neuropsychiatric illness. Here, we investigated neural phase signaling in two mouse models of cognitive dysfunction: mice with genetically induced hyperdopaminergia [dopamine transporter knock-out (DAT-KO) mice] and mice with genetically induced NMDA receptor hypofunction [NMDA receptor subunit-1 knockdown (NR1-KD) mice]. Cognitive function in these mice was assessed using a radial-arm maze task, and local field potentials were recorded from dorsal hippocampus and prefrontal cortex as DAT-KO mice, NR1-KD mice, and their littermate controls engaged in behavioral exploration. Our results demonstrate that both DAT-KO and NR1-KD mice display deficits in spatial cognitive performance. Moreover, we show that persistent hyperdopaminergia alters interstructural phase signaling, whereas NMDA receptor hypofunction alters interstructural and intrastructural phase signaling. These results demonstrate that dopamine and NMDA receptor dependent glutamate signaling play a critical role in coordinating neural phase signaling, and encourage further studies to investigate the role that deficits in phase signaling play in mediating cognitive dysfunction. PMID- 19553462 TI - Suppression of spreading depression-like events in locusts by inhibition of the NO/cGMP/PKG pathway. AB - Despite considerable research attention focused on mechanisms underlying neural spreading depression (SD), because of its association with important human CNS pathologies, such as stroke and migraine, little attention has been given to explaining its occurrence and regulation in invertebrates. In the locust metathoracic ganglion (MTG), an SD-like event occurs during heat and anoxia stress, which results in cessation of neuronal output for the duration of the applied stress. SD-like events were characterized by an abrupt rise in extracellular potassium ion concentration ([K(+)](o)) from a baseline concentration of approximately 8 to >30 mm, which returned to near baseline concentrations after removal of the applied stress. After return to baseline [K(+)](o), neuronal output (ventilatory motor pattern activity) from the MTG recovered. Unlike mammalian neurons, which depolarize almost completely during SD, locust neurons only partially depolarized. SD-like events in the locust CNS were suppressed by pharmacological inhibition of the nitric oxide/cyclic guanosine monophosphate/protein kinase G (NO/cGMP/PKG) pathway and were exacerbated by its activation. Also, environmental stressors such as heat and anoxia increased production of nitric oxide in the locust CNS. Finally, for the intact animal, manipulation of the pathway affected the speed of recovery from suffocation by immersion under water. We propose that SD-like events in locusts provide an adaptive mechanism for surviving extreme environmental conditions. The highly conserved nature of the NO/cGMP/PKG signaling pathway suggests that it may be involved in modulating SD in other organisms, including mammals. PMID- 19553463 TI - FOXO3a is broadly neuroprotective in vitro and in vivo against insults implicated in motor neuron diseases. AB - Aging is a risk factor for the development of adult-onset neurodegenerative diseases. Although some of the molecular pathways regulating longevity and stress resistance in lower organisms are defined (i.e., those activating the transcriptional regulators DAF-16 and HSF-1 in Caenorhabditis elegans), their relevance to mammals and disease susceptibility are unknown. We studied the signaling controlled by the mammalian homolog of DAF-16, FOXO3a, in model systems of motor neuron disease. Neuron death elicited in vitro by excitotoxic insult or the expression of mutant SOD1, mutant p150(glued), or polyQ-expanded androgen receptor was abrogated by expression of nuclear-targeted FOXO3a. We identify a compound [Psammaplysene A (PA)] that increases nuclear localization of FOXO3a in vitro and in vivo and show that PA also protects against these insults in vitro. Administration of PA to invertebrate model systems of neurodegeneration similarly blocked neuron death in a DAF-16/FOXO3a-dependent manner. These results indicate that activation of the DAF-16/FOXO3a pathway, genetically or pharmacologically, confers protection against the known causes of motor neuron diseases. PMID- 19553464 TI - NOS1AP regulates dendrite patterning of hippocampal neurons through a carboxypeptidase E-mediated pathway. AB - During neuronal development, neurons form elaborate dendritic arbors that receive signals from axons. Additional studies are needed to elucidate the factors regulating the establishment of dendritic patterns. Our work explored possible roles played by nitric oxide synthase 1 adaptor protein (NOS1AP; also known as C terminal PDZ ligand of neuronal nitric oxide synthase or CAPON) in dendritic patterning of cultured hippocampal neurons. Here we report that the long isoform of NOS1AP (NOS1AP-L) plays a novel role in regulating dendrite outgrowth and branching. NOS1AP-L decreases dendrite number when overexpressed at any interval between day in vitro (DIV) 0 and DIV 12, and knockdown of NOS1AP-L results in increased dendrite number. In contrast, the short isoform of NOS1AP (NOS1AP-S) decreases dendrite number only when overexpressed during DIV 5-7. Using mutants of NOS1AP-L, we show that neither the PDZ-binding domain nor the PTB domain is necessary for the effects of NOS1AP-L. We have functionally narrowed the region of NOS1AP-L that mediates this effect to the middle amino acids 181-307, a region that is not present in NOS1AP-S. Furthermore, we performed a yeast two-hybrid screen and identified carboxypeptidase E (CPE) as a binding partner for the middle region of NOS1AP-L. Biochemical and cellular studies reveal that CPE mediates the effects of NOS1AP on dendrite morphology. Together, our results suggest that NOS1AP-L plays an important role in the initiation, outgrowth, and maintenance of dendrites during development. PMID- 19553465 TI - Inhibition of the mammalian target of rapamycin signaling pathway suppresses dentate granule cell axon sprouting in a rodent model of temporal lobe epilepsy. AB - Dentate granule cell axon (mossy fiber) sprouting is a common abnormality in patients with temporal lobe epilepsy. Mossy fiber sprouting creates an aberrant positive-feedback network among granule cells that does not normally exist. Its role in epileptogenesis is unclear and controversial. If it were possible to block mossy fiber sprouting from developing after epileptogenic treatments, its potential role in the pathogenesis of epilepsy could be tested. Previous attempts to block mossy fiber sprouting have been unsuccessful. The present study targeted the mammalian target of rapamycin (mTOR) signaling pathway, which regulates cell growth and is blocked by rapamycin. Rapamycin was focally, continuously, and unilaterally infused into the dorsal hippocampus for prolonged periods beginning within hours after rats sustained pilocarpine-induced status epilepticus. Infusion for 1 month reduced aberrant Timm staining (a marker of mossy fibers) in the granule cell layer and molecular layer. Infusion for 2 months inhibited mossy fiber sprouting more. However, after rapamycin infusion ceased, aberrant Timm staining developed and approached untreated levels. When onset of infusion began after mossy fiber sprouting had developed for 2 months, rapamycin did not reverse aberrant Timm staining. These findings suggest that inhibition of the mTOR signaling pathway suppressed development of mossy fiber sprouting. However, suppression required continual treatment, and rapamycin treatment did not reverse already established axon reorganization. PMID- 19553466 TI - Selecting for memory? The influence of selective attention on the mnemonic binding of contextual information. AB - Not all of what is experienced is remembered later. Behavioral evidence suggests that the manner in which an event is processed influences which aspects of the event will later be remembered. The present experiment investigated the neural correlates of "selective encoding," or the mechanisms that support the encoding of some elements of an event in preference to others. Event-related MRI data were acquired while volunteers selectively attended to one of two different contextual features of study items (color or location). A surprise memory test for the items and both contextual features was subsequently administered to determine the influence of selective attention on the neural correlates of contextual encoding. Activity in several cortical regions indexed later memory success selectively for color or location information, and this encoding-related activity was enhanced by selective attention to the relevant feature. Critically, a region in the hippocampus responded selectively to attended source information (whether color or location), demonstrating encoding-related activity for attended but not for nonattended source features. Together, the findings suggest that selective attention modulates the magnitude of activity in cortical regions engaged by different aspects of an event, and hippocampal encoding mechanisms seem to be sensitive to this modulation. Thus, the information that is encoded into a memory representation is biased by selective attention, and this bias is mediated by cortical-hippocampal interactions. PMID- 19553467 TI - Evidence of action sequence chunking in goal-directed instrumental conditioning and its dependence on the dorsomedial prefrontal cortex. AB - The current study investigated the contribution of the dorsomedial prefrontal cortex (dmPFC) to instrumental action selection. We found that cell body lesions of the dmPFC, centered on the medial agranular area, spared rats' ability to choose between actions based on either the value or the discriminative stimulus properties of an outcome. We next examined the effects of these lesions on action sequence learning using a concurrent bidirectional heterogeneous chain task in which the identity of the reward delivered was determined by the order in which the two lever press actions were performed. Although both lesioned rats and sham controls learned to perform the task, we found that they relied on different behavioral strategies to do so. In subsequent tests, rats in the sham group were able to withhold their performance of a sequence when either its associated outcome was devalued or the contingency between that sequence and its outcome was degraded by delivering the outcome noncontingently. Interestingly, lesioned rats failed to reorganize their performance at the action sequence level and, rather, were found to withhold their performance of the terminal response in the sequence that had earned the devalued outcome relative to the more distal response, suggesting that they represented the elements of the sequence as distinct behavioral units. These findings demonstrate that rats can use sequence-level representations, or action chunks, to organize their behavior in a goal-directed manner and indicate that the dmPFC plays a critical role in this process. PMID- 19553468 TI - Histone deacetylases 1 and 2 form a developmental switch that controls excitatory synapse maturation and function. AB - The structural assembly of synapses can be accomplished in a rapid time frame, although most nascent synapses formed during early development are not fully functional and respond poorly to presynaptic action potentials. The mechanisms that are responsible for this delay in synapse maturation are unknown. Histone deacetylases (HDACs) regulate the activity state of chromatin and repress gene expression through the removal of acetyl groups from histones. Class I HDACs, which include HDAC1 and HDAC2, are expressed in the CNS, although their specific role in neuronal function has not been studied. To delineate the contribution of HDAC1 and HDAC2 in the brain, we have used pharmacological inhibitors of HDACs and mice with conditional alleles to HDAC1 and HDAC2. We found that a decrease in the activities of both HDAC1 and HDAC2 during early synaptic development causes a robust facilitation of excitatory synapse maturation and a modest increase in synapse numbers. In contrast, in mature neurons a decrease in HDAC2 levels alone was sufficient to attenuate basal excitatory neurotransmission without a significant change in the numbers of detectable nerve terminals. Therefore, we propose that HDAC1 and HDAC2 form a developmental switch that controls synapse maturation and function acting in a manner dependent on the maturational states of neuronal networks. PMID- 19553469 TI - Human Golgi antiapoptotic protein modulates intracellular calcium fluxes. AB - Golgi antiapoptotic protein (GAAP) is a novel regulator of cell death that is highly conserved in eukaryotes and present in some poxviruses, but its molecular mechanism is unknown. Given that alterations in intracellular Ca(2+) homeostasis play an important role in determining cell sensitivity to apoptosis, we investigated if GAAP affected Ca(2+) signaling. Overexpression of human (h)-GAAP suppressed staurosporine-induced, capacitative Ca(2+) influx from the extracellular space. In addition, it reduced histamine-induced Ca(2+) release from intracellular stores through inositol trisphosphate receptors. h-GAAP not only decreased the magnitude of the histamine-induced Ca(2+) fluxes from stores to cytosol and mitochondrial matrices, but it also reduced the induction and frequency of oscillatory changes in cytosolic Ca(2+). Overexpression of h-GAAP lowered the Ca(2+) content of the intracellular stores and decreased the efficacy of IP(3), providing possible explanations for the observed results. Opposite effects were obtained when h-GAAP was knocked down by siRNA. Thus, our data demonstrate that h-GAAP modulates intracellular Ca(2+) fluxes induced by both physiological and apoptotic stimuli. PMID- 19553470 TI - Clustering and lateral concentration of raft lipids by the MAL protein. AB - MAL, a compact hydrophobic, four-transmembrane-domain apical protein that copurifies with detergent-resistant membranes is obligatory for the machinery that sorts glycophosphatidylinositol (GPI)-anchored proteins and others to the apical membrane in epithelia. The mechanism of MAL function in lipid-raft mediated apical sorting is unknown. We report that MAL clusters formed by two independent procedures-spontaneous clustering of MAL tagged with the tandem dimer DiHcRED (DiHcRED-MAL) in the plasma membrane of COS7 cells and antibody-mediated cross-linking of FLAG-tagged MAL-laterally concentrate markers of sphingolipid rafts and exclude a fluorescent analogue of phosphatidylethanolamine. Site directed mutagenesis and bimolecular fluorescence complementation analysis demonstrate that MAL forms oligomers via xx intramembrane protein-protein binding motifs. Furthermore, results from membrane modulation by using exogenously added cholesterol or ceramides support the hypothesis that MAL-mediated association with raft lipids is driven at least in part by positive hydrophobic mismatch between the lengths of the transmembrane helices of MAL and membrane lipids. These data place MAL as a key component in the organization of membrane domains that could potentially serve as membrane sorting platforms. PMID- 19553471 TI - Cadherin adhesion receptors orient the mitotic spindle during symmetric cell division in mammalian epithelia. AB - Oriented cell division is a fundamental determinant of tissue organization. Simple epithelia divide symmetrically in the plane of the monolayer to preserve organ structure during epithelial morphogenesis and tissue turnover. For this to occur, mitotic spindles must be stringently oriented in the Z-axis, thereby establishing the perpendicular division plane between daughter cells. Spatial cues are thought to play important roles in spindle orientation, notably during asymmetric cell division. The molecular nature of the cortical cues that guide the spindle during symmetric cell division, however, is poorly understood. Here we show directly for the first time that cadherin adhesion receptors are required for planar spindle orientation in mammalian epithelia. Importantly, spindle orientation was disrupted without affecting tissue cohesion or epithelial polarity. This suggests that cadherin receptors can serve as cues for spindle orientation during symmetric cell division. We further show that disrupting cadherin function perturbed the cortical localization of APC, a microtubule interacting protein that was required for planar spindle orientation. Together, these findings establish a novel morphogenetic function for cadherin adhesion receptors to guide spindle orientation during symmetric cell division. PMID- 19553472 TI - The low-affinity receptor for neurotrophins p75NTR plays a key role for satellite cell function in muscle repair acting via RhoA. AB - Regeneration of muscle fibers, lost during pathological muscle degeneration or after injuries, is mediated by the production of new myofibres. This process, sustained by the resident stem cells of the muscle, the satellite cells, is finely regulated by local cues, in particular by cytokines and growth factors. Evidence in the literature suggests that nerve growth factor (NGF) is involved in muscle fiber regeneration; however, its role and mechanism of action were unclear. We have investigated this issue in in vivo mouse models of muscle regeneration and in primary myogenic cells. Our results demonstrate that NGF acts through its low-affinity receptor p75(NTR) in a developmentally regulated signaling pathway necessary to myogenic differentiation and muscle repair in vivo. We also demonstrate that this action of NGF is mediated by the down regulation of RhoA-GTP signaling in myogenic cells. PMID- 19553473 TI - Interaction of CDK5RAP2 with EB1 to track growing microtubule tips and to regulate microtubule dynamics. AB - Mutations in cdk5rap2 are linked to autosomal recessive primary microcephaly, and attention has been paid to its function at centrosomes. In this report, we demonstrate that CDK5RAP2 localizes to microtubules and concentrates at the distal tips in addition to centrosomal localization. CDK5RAP2 interacts directly with EB1, a prototypic member of microtubule plus-end tracking proteins, and contains the basic and Ser-rich motif responsible for EB1 binding. The EB1 binding motif is conserved in the CDK5RAP2 sequences of chimpanzee, bovine, and dog but not in those of rat and mouse, suggesting a function gained during the evolution of mammals. The mutation of the Ile/Leu-Pro dipeptide within the motif abolishes EB1 interaction and plus-end attachment. In agreement with the mutational analysis, suppression of EB1 expression inhibits microtubule tip tracking of CDK5RAP2. We have also found that the CDK5RAP2-EB1 complex regulates microtubule dynamics and stability. CDK5RAP2 depletion by RNA interference impacts the dynamic behaviors of microtubules. The CDK5RAP2-EB1 complex induces microtubule bundling and acetylation when expressed in cell cultures and stimulates microtubule assembly and bundle formation in vitro. Collectively, these results show that CDK5RAP2 targets growing microtubule tips in association with EB1 to regulate microtubule dynamics. PMID- 19553474 TI - {alpha}-synuclein and its A30P mutant affect actin cytoskeletal structure and dynamics. AB - The function of alpha-synuclein, a soluble protein abundant in the brain and concentrated at presynaptic terminals, is still undefined. Yet, alpha-synuclein overexpression and the expression of its A30P mutant are associated with familial Parkinson's disease. Working in cell-free conditions, in two cell lines as well as in primary neurons we demonstrate that alpha-synuclein and its A30P mutant have different effects on actin polymerization. Wild-type alpha-synuclein binds actin, slows down its polymerization and accelerates its depolymerization, probably by monomer sequestration; A30P mutant alpha-synuclein increases the rate of actin polymerization and disrupts the cytoskeleton during reassembly of actin filaments. Consequently, in cells expressing mutant alpha-synuclein, cytoskeleton dependent processes, such as cell migration, are inhibited, while exo- and endocytic traffic is altered. In hippocampal neurons from mice carrying a deletion of the alpha-synuclein gene, electroporation of wild-type alpha synuclein increases actin instability during remodeling, with growth of lamellipodia-like structures and apparent cell enlargement, whereas A30P alpha synuclein induces discrete actin-rich foci during cytoskeleton reassembly. In conclusion, alpha-synuclein appears to play a major role in actin cytoskeletal dynamics and various aspects of microfilament function. Actin cytoskeletal disruption induced by the A30P mutant might alter various cellular processes and thereby play a role in the pathogenesis of neurodegeneration. PMID- 19553476 TI - Differential effects of cognitive demand on human cortical activation associated with vibrotactile stimulation. AB - This event-related functional MRI study examines the neural correlates of vibrotactile sensation within the context of different psychophysical demands. Nine subjects received vibrotactile stimuli on the right volar forearm during detection, localization, and passive tasks. In the detection task, subjects indicated the offset (end) of each stimulus by pressing a response key with their left hand. In the localization task, subjects identified the location of the stimulus ("distal?" or "proximal?") by pressing the appropriate response key 4 s after the end of the stimulus. In the passive task, subjects received the same vibrotactile stimuli, but no response was required. Analysis of stimulus-evoked activity compared with the resting baseline period revealed significant bilateral secondary somatosensory cortex activation for all three tasks. However, only in the offset-detection and localization tasks was stimulus-evoked activation observed in other expected areas of tactile processing, such as contralateral primary somatosensory cortex neighboring the posterior parietal cortex (SI/PPC) and in bilateral anterior insular cortex (aIC). During the localization task, we identified vibrotactile-evoked activation in the right aIC, which was maintained after the termination of the stimulus. Results suggest that vibrotactile-related activation within SI/PPC and aIC is enhanced by the increased levels of attention and cognitive demands required by the detection and localization tasks. Activation of aIC not only during vibrotactile stimulation, but also during the poststimulus delay in the localization trials, is consistent with the growing literature linking this area with the perception and short-term memory of tactile information. PMID- 19553475 TI - Regulation of the benzamil-insensitive salt taste receptor by intracellular Ca2+, protein kinase C, and calcineurin. AB - The regulation of the benzamil (Bz)-insensitive salt taste receptor was investigated by intracellular Ca2+ ([Ca2+]i), protein kinase C (PKC), and the Ca2+-dependent serine-threonine phosphatase, calcineurin (PP2B), by monitoring chorda tympani taste nerve responses to 0.1 M NaCl solutions containing Bz (5x10( 6) M) and resiniferatoxin (RTX; 0-10x10(-6) M) in Sprague-Dawley rats and in wild type (WT) and transient receptor potential vanilloid-1 knockout (TRPV1 KO) mice. In rats and WT mice, RTX increased the NaCl+Bz chorda tympani responses between 0.25x10(-6) and 1x10(-6) M and inhibited the responses above 1x10(-6) M. Decreasing taste receptor cell (TRC) [Ca2+]i with BAPTA loading, activation of PKC with 4alpha-phorbol-12,13-didecanoate (PMA), or inhibition of PP2B by cyclosporin A or FK-506, enhanced the magnitude of the Bz-insensitive NaCl chorda tympani responses in the presence of RTX and either minimized or completely eliminated the decrease in the chorda tympani response>1x10(-6) M RTX. In contrast, increasing TRC [Ca2+]i with ionomycin inhibited Bz-insensitive NaCl chorda tympani responses in the presence of RTX. No effect of the cited modulators was observed on the chorda tympani responses in WT mice and rats in the presence of TRPV1 blocker SB-366791 (1x10(-6) M) or in TRPV1 KO mice. 32P labeling demonstrated direct phosphorylation of TRPV1 or TRPV1t in anterior lingual epithelium by PMA, cyclosporin A, or FK-506. PMA also enhanced the RTX sensitive unilateral apical Na+ flux in polarized fungiform TRC in vitro. We conclude that TRPV1 or its variant TRPV1t is phosphorylated and dephosphorylated by PKC and PP2B, respectively, and either sensitizes or desensitizes the Bz insensitive NaCl chorda tympani responses to RTX stimulation. PMID- 19553478 TI - Multijoint reflex responses to constant-velocity volitional movements of the stroke elbow. AB - Multijoint reflex coupling could impact the voluntary control of functional arm movements in people post stroke. The multijoint responses to active-assist, constant-velocity movements of the elbow joint were measured in 14 individuals post stroke and 9 neurologically intact controls. Resulting responses in the stroke group illustrated a change in the reflex coupling of the elbow and shoulder muscles compared with passive perturbations of the spastic elbow. Voluntary effort during constant-velocity elbow extension resulted in reflex shoulder abduction, differing from the reflex coupling observed between the elbow flexors and shoulder adductors observed during passive elbow extension. These results suggest that post stroke, voluntary drive alters reflex coupling of the elbow and shoulder. Flexion of the elbow during active-assist also resulted in reflex coupling. Shoulder abduction torque decreased with constant-velocity flexion of the elbow; however, no net adduction was observed at the end of the perturbation. Shoulder flexion/extension and internal/external rotation torque responses demonstrated similar modulations to imposed active-assist perturbations of the elbow in subjects post stroke. Responses were absent during passive perturbations of the control elbow; however, shoulder torque modulations were observed during constant-velocity, active-assist tasks. The active-assist response patterns in controls were similar to stroke subjects during the extension task but opposite during flexion of the elbow. This study provides evidence of a neural coupling between elbow and shoulder muscles and a modulation of this coupling during voluntary drive of the spastic arm. PMID- 19553477 TI - Brain stem feedback in a computational model of birdsong sequencing. AB - Uncovering the roles of neural feedback in the brain is an active area of experimental research. In songbirds, the telencephalic premotor nucleus HVC receives neural feedback from both forebrain and brain stem areas. Here we present a computational model of birdsong sequencing that incorporates HVC and associated nuclei and builds on the model of sparse bursting presented in our preceding companion paper. Our model embodies the hypotheses that 1) different networks in HVC control different syllables or notes of birdsong, 2) interneurons in HVC not only participate in sparse bursting but also provide mutual inhibition between networks controlling syllables or notes, and 3) these syllable networks are sequentially excited by neural feedback via the brain stem and the afferent thalamic nucleus Uva, or a similar feedback pathway. We discuss the model's ability to unify physiological, behavioral, and lesion results and we use it to make novel predictions that can be tested experimentally. The model suggests a neural basis for sequence variations, shows that stimulation in the feedback pathway may have different effects depending on the balance of excitation and inhibition at the input to HVC from Uva, and predicts deviations from uniform expansion of syllables and gaps during HVC cooling. PMID- 19553480 TI - Slow oscillation state facilitates epileptiform events in the hippocampus. AB - In mesial temporal lobe (MTL) epilepsy, which typically involves the hippocampus (HPC), epileptiform events are enhanced during slow wave sleep (SWS). It remains unclear how and why the electroencephalographic (EEG) states that constitute SWS might predispose the HPC to this type of pathological activity. Recently our laboratory has described a novel state of deactivated hippocampal EEG activity that occurs during both SWS and urethan anesthesia: the slow oscillation (SO). This activity is characterized by a high-amplitude approximately 1-Hz signal, high synchrony within the hippocampus, and a dynamic coordination with neocortical SO. To assess how this activity state might influence epileptiform discharges, we studied the properties of stimulation-evoked and spontaneous epileptiform events elicited in the HPC of urethan-anesthetized rats. We compared those elicited during the SO to those occurring during the theta rhythm. The average duration but not the amplitude of evoked afterdischarges (ADs) was consistently larger during the SO. In addition, spontaneous epileptiform events were more frequent and of higher amplitude during the SO. Last, the bilateral propagation of both ADs and spontaneous events in the hippocampus was enhanced during the SO. These results imply that the threshold for the generation and propagation of epileptiform activity in the hippocampus is lowered during the SO and that this state may be a seed for the initiation, maintenance, and generalization of MTL epilepsy. Further examination of the pathophysiology of sleep-epilepsy interactions in the HPC will be of benefit for an understanding of the mechanisms, prognosis, and therapy for this form of epilepsy. PMID- 19553479 TI - Response linearity of alert monkey non-eye movement vestibular nucleus neurons during sinusoidal yaw rotation. AB - Vestibular afferents display linear responses over a range of amplitudes and frequencies, but comparable data for central vestibular neurons are lacking. To examine the effect of stimulus frequency and magnitude on the response sensitivity and linearity of non-eye movement central vestibular neurons, we recorded from the vestibular nuclei in awake rhesus macaques during sinusoidal yaw rotation at frequencies between 0.1 and 2 Hz and between 7.5 and 210 degrees/s peak velocity. The dynamics of the neurons' responses across frequencies, while holding peak velocity constant, was consistent with previous studies. However, as the peak velocity was varied, while holding the frequency constant, neurons demonstrated lower sensitivities with increasing peak velocity, even at the lowest peak velocities tested. With increasing peak velocity, the proportion of neurons that silenced during a portion of the response increased. However, the decrease in sensitivity of these neurons with higher peak velocities of rotation was not due to increased silencing during the inhibitory portion of the cycle. Rather the neurons displayed peak firing rates that did not increase in proportion to head velocity as the peak velocity of rotation increased. These data suggest that, unlike vestibular afferents, the central vestibular neurons without eye movement sensitivity examined in this study do not follow linear systems principles even at low velocities. PMID- 19553481 TI - Neural activity in the human brain signals logical rule identification. AB - To select an appropriate action, we conform to a behavioral rule determined uniquely in each behavioral context. If the rule is not predetermined and must be discovered, we often test hypotheses concerning rules by applying one candidate rule after another. The neural mechanisms underlying such rule identification are still unknown. To explore which brain areas are involved in the process of logical rule identification and to determine whether such areas differ from those taking part in implementing the rule to find a suitable action, we measured brain activation using functional magnetic resonance imaging while subjects performed a rule-identification task. The subjects were required to select a red or blue square on a screen based on either a "sequence rule" or a "probability rule." Positive or negative feedback to the subject's choice led the subject to identify the correct rule. We found that the posterior medial frontal cortex (pMFC), caudate nucleus, fusiform gyrus, and middle temporal cortex exhibited significant activation during the period when subjects underwent the hypothesis testing. Among these brain areas, the pMFC and caudate nucleus were also activated in response to the critical feedback signals selectively during the trials when the subjects identified a rule. Furthermore, we found a significant enhancement in effective connectivity between the active regions in the pMFC and caudate regions. PMID- 19553482 TI - Dendritic calcium channels and their activation by synaptic signals in auditory coincidence detector neurons. AB - The avian nucleus laminaris (NL) encodes the azimuthal location of low-frequency sound sources by detecting the coincidence of binaural signals. Accurate coincidence detection requires precise developmental regulation of the lengths of the fine, bitufted dendrites that characterize neurons in NL. Such regulation has been suggested to be driven by local, synaptically mediated, dendritic signals such as Ca(2+). We examined Ca(2+) signaling through patch clamp and ion imaging experiments in slices containing nucleus laminaris from embryonic chicks. Voltage clamp recordings of neurons located in the NL showed the presence of large Ca(2+) currents of two types, a low voltage-activated, fast inactivating Ni(2+) sensitive channel resembling mammalian T-type channels, and a high voltage activated, slowly inactivating Cd(2+) sensitive channel. Two-photon Ca(2+) imaging showed that both channel types were concentrated on dendrites, even at their distal tips. Single action potentials triggered synaptically or by somatic current injection immediately elevated Ca(2+) throughout the entire cell. Ca(2+) signals triggered by subthreshold synaptic activity were highly localized. Thus when electrical activity is suprathreshold, Ca(2+) channels ensure that Ca(2+) rises in all dendrites, even those that are synaptically inactive. PMID- 19553483 TI - Presynaptic mechanisms of endocannabinoid-mediated long-term depression in the hippocampus. AB - Endogenous cannabinoids (eCBs) mobilized from postsynaptic cells activate presynaptic cannabinoid receptors and thereby inhibit synaptic transmitter release. The inhibition can be either brief (seconds) or long-lasting (minutes to hours). Recent studies provide insight into the presynaptic mechanisms responsible for long-lasting, eCB-mediated long-term depression. Among these, the proteins PKA and RIM1alpha appear to be crucial, although other possibilities are emerging. PMID- 19553484 TI - Chemosensory responses to CO2 in multiple brain stem nuclei determined using a voltage-sensitive dye in brain slices from rats. AB - We used epifluorescence microscopy and a voltage-sensitive dye, di-8-ANEPPS, to study changes in membrane potential during hypercapnia with or without synaptic blockade in chemosensory brain stem nuclei: the locus coeruleus (LC), the nucleus of the solitary tract, lateral paragigantocellularis nucleus, raphe pallidus, and raphe obscurus and, in putative nonchemosensitive nuclei, the gigantocellularis reticular nucleus and the spinotrigeminal nucleus. We studied the response to hypercapnia in LC cells to evaluate the performance characteristics of the voltage-sensitive dye. Hypercapnia depolarized many LC cells and the voltage responses to hypercapnia were diminished, but not eradicated, by synaptic blockade (there were intrinsically CO2-sensitive cells in the LC). The voltage response to hypercapnia was substantially diminished after inhibiting fast Na+ channels with tetrodotoxin. Thus action potential-related activity was responsible for most of the optical signal that we detected. We systematically examined CO2 sensitivity among cells in brain stem nuclei to test the hypothesis that CO2 sensitivity is a ubiquitous phenomenon, not restricted to nominally CO2 chemosensory nuclei. We found intrinsically CO2 sensitive neurons in all the nuclei that we examined; even the nonchemosensory nuclei had small numbers of intrinsically CO2 sensitive neurons. However, synaptic blockade significantly altered the distribution of CO2-sensitive cells in all of the nuclei so that the cellular response to CO2 in more intact preparations may be difficult to predict based on studies of intrinsic neuronal activity. Thus CO2-sensitive neurons are widely distributed in chemosensory and nonchemosensory nuclei and CO2 sensitivity is dependent on inhibitory and excitatory synaptic activity even within brain slices. Neuronal CO2 sensitivity important for the behavioral response to CO2 in intact animals will thus be determined as much by synaptic mechanisms and patterns of connectivity throughout the brain as by intrinsic CO2 sensitivity. PMID- 19553485 TI - The influence of neuroactive steroid lipophilicity on GABAA receptor modulation: evidence for a low-affinity interaction. AB - Anesthetic steroids with actions at gamma-aminobutyric acid type A receptors (GABA(A)Rs) may access transmembrane domain binding site(s) directly from the plasma cell membrane. Accordingly, the effective concentration in lipid phase and the ability of the steroid to meet pharmacophore requirements for activity will both contribute to observed steady-state potency. Furthermore, onset and offset of receptor effects may be rate limited by lipid partitioning. Here we show that several GABA-active steroids, including naturally occurring neurosteroids, of different lipophilicity differ in kinetics and potency at GABA(A)Rs. The hydrophobicity ranking predicted relative potency of GABA(A)R potentiation and predicted current offset kinetics. Kinetic offset differences among steroids were largely eliminated by gamma-cyclodextrin, a scavenger of unbound steroid, suggesting that affinity differences among the analogues are dwarfed by the contributions of nonspecific accumulation. A 7-nitrobenz-2-oxa-1,3-diazole (NBD) tagged fluorescent analogue of the low-lipophilicity alphaxalone (C17-NBD alphaxalone) exhibited faster nonspecific accumulation and departitioning than those of a fluorescent analogue of the high-lipophilicity (3alpha,5alpha)-3 hydroxypregnan-20-one (C17-NBD-3alpha5alphaA). These differences were paralleled by differences in potentiation of GABA(A)R function. The enantiomer of C17-NBD 3alpha5alphaA, which does not satisfy pharmacophore requirements for steroid potentiation, exhibited identical fluorescence kinetics and distribution to C17 NBD-3alpha5alphaA, but was inactive at GABA(A)Rs. Simple simulations supported our major findings, which suggest that neurosteroid binding affinity is low. Therefore both specific (e.g., fulfilling pharmacophore requirements) and nonspecific (e.g., lipid solubility) properties contribute to the potency and longevity of anesthetic steroid action. PMID- 19553487 TI - TNFalpha mechanically sensitizes masseter muscle afferent fibers of male rats. AB - Behavioral evidence in rats indicates that injection of tumor necrosis factor alpha (TNFalpha) into skeletal muscle results in a prolonged mechanical sensitization without gross inflammation. To investigate whether a peripheral mechanism could underlie this effect, in the present study, TNFalpha (1 or 0.1 microg) was injected into the rat masseter muscle to assess its effect on the excitability and mechanical threshold (MT) of muscle nociceptors as well as on inflammation. Expression of TNFR1 (P55 receptors) and TNFR2 (P75 receptors) by the masseter muscle and trigeminal ganglion neurons that innervate that muscle was determined by Western blot and immunohistochemistry, respectively. The Evans blue dye technique was used at the end of the TNFalpha experiments to assess for plasma protein extravasation. In subsequent experiments to confirm the involvement of receptor activation in TNFalpha-induced effects, P55 or P75 receptor antibody was co-injected with TNFalpha. Intramuscular injection of 1 microg TNFalpha did not excite nociceptors but did significantly decrease MT compared with vehicle control. There was no evidence of gross inflammation 3 h after injection of TNFalpha. Co-injection of TNFalpha with P55 or P75 receptor antibodies attenuated TNFalpha-induced mechanical sensitization. P55 and P75 receptors were expressed by 29 and 62% of masseter nociceptors, respectively. These findings indicate that TNFalpha induces mechanical sensitization of masseter nociceptors that is mediated through activation of peripheral P55 and P75 receptors. These results support the hypothesis that a peripheral receptor mechanism could contribute to TNFalpha-induced noninflammatory mechanical sensitization of skeletal muscle previously reported in behaving rats. PMID- 19553486 TI - GABA shapes selectivity for the rate and direction of frequency-modulated sweeps in the auditory cortex. AB - In the pallid bat auditory cortex and inferior colliculus (IC), the majority of neurons tuned in the echolocation range is selective for the direction and rate of frequency-modulated (FM) sweeps used in echolocation. Such selectivity is shaped mainly by spectrotemporal asymmetries in sideband inhibition. An early arriving, low-frequency inhibition (LFI) shapes direction selectivity. A delayed, high-frequency inhibition (HFI) shapes rate selectivity for downward sweeps. Using iontophoretic blockade of GABAa receptors, we show that cortical FM sweep selectivity is at least partially shaped locally. GABAa receptor antagonists, bicuculline or gabazine, reduced or eliminated direction and rate selectivity in approximately 50% of neurons. Intracortical GABA shapes FM sweep selectivity by either creating the underlying sideband inhibition or by advancing the arrival time of inhibition relative to excitation. Given that FM sweep selectivity and asymmetries in sideband inhibition are already present in the IC, these data suggest a refinement or recreation of similar response properties at the cortical level. PMID- 19553489 TI - Seasonal plasticity of auditory saccular sensitivity in the vocal plainfin midshipman fish, Porichthys notatus. AB - The plainfin midshipman fish, Porichthys notatus, is a seasonally breeding species of marine teleost fish that generates acoustic signals for intraspecific social and reproductive-related communication. Female midshipman use the inner ear saccule as the main acoustic endorgan for hearing to detect and locate vocalizing males that produce multiharmonic advertisement calls during the breeding season. Previous work showed that the frequency sensitivity of midshipman auditory saccular afferents changed seasonally with female reproductive state such that summer reproductive females became better suited than winter nonreproductive females to encode the dominant higher harmonics of the male advertisement calls. The focus of this study was to test the hypothesis that seasonal reproductive-dependent changes in saccular afferent tuning is paralleled by similar changes in saccular sensitivity at the level of the hair cell receptor. Here, I examined the evoked response properties of midshipman saccular hair cells from winter nonreproductive and summer reproductive females to determine if reproductive state affects the frequency response and threshold of the saccule to behaviorally relevant single tone stimuli. Saccular potentials were recorded from populations of hair cells in vivo while sound was presented by an underwater speaker. Results indicate that saccular hair cells from reproductive females had thresholds that were approximately 8 to 13 dB lower than nonreproductive females across a broad range of frequencies that included the dominant higher harmonic components and the fundamental frequency of the male's advertisement call. These seasonal-reproductive-dependent changes in thresholds varied differentially across the three (rostral, middle, and caudal) regions of the saccule. Such reproductive-dependent changes in saccule sensitivity may represent an adaptive plasticity of the midshipman auditory sense to enhance mate detection, recognition, and localization during the breeding season. PMID- 19553490 TI - How reliable is the pattern adaptation technique? A modeling study. AB - Upon prolonged viewing of a sinusoidal grating, the visual system is selectively desensitized to the spatial frequency of the grating, while the sensitivity to other spatial frequencies remains largely unaffected. This technique, known as pattern adaptation, has been so central to the psychophysical study of the mechanisms of spatial vision that it is sometimes referred to as the "psychologist's microelectrode." While this approach implicitly assumes that the adaptation behavior of the system is diagnostic of the corresponding underlying neural mechanisms, this assumption has never been explicitly tested. We tested this assumption using adaptation bandwidth, or the range of spatial frequencies affected by adaptation, as a representative measure of adaptation. We constructed an intentionally simple neuronal ensemble model of spatial frequency processing and examined the extent to which the adaptation bandwidth at the system level reflected the bandwidth at the neuronal level. We find that the adaptation bandwidth could vary widely even when all spatial frequency tuning parameters were held constant. Conversely, different spatial frequency tuning parameters were able to elicit similar adaptation bandwidths from the neuronal ensemble. Thus, the tuning properties of the underlying units did not reliably reflect the adaptation bandwidth at the system level, and vice versa. Furthermore, depending on the noisiness of adaptation at the neural level, the same neuronal ensemble was able to produce selective or nonselective adaptation at the system level, indicating that a lack of selective adaptation at the system level cannot be taken to mean a lack of tuned mechanisms at the neural level. Together, our results indicate that pattern adaptation cannot be used to reliably estimate the tuning properties of the underlying units, and imply, more generally, that pattern adaptation is not a reliable tool for studying the neural mechanisms of pattern analysis. PMID- 19553488 TI - Serotonergic modulation of the trigeminocardiac reflex neurotransmission to cardiac vagal neurons in the nucleus ambiguus. AB - Stimulation of the trigeminal nerve evokes a dramatic decrease in heart rate and blood pressure, and this reflex has generally been termed the trigeminocardiac reflex. A subset of the trigeminocardiac reflex is the diving reflex in which the nasal mucosa is stimulated with water or air-borne chemical irritants. Activation of the diving reflex evokes a pronounced bradycardia, mediated by increased parasympathetic cardiac activity, and is the most powerful autonomic reflex. However, exaggeration of this protective response could be detrimental and has been implicated in Sudden Infant Death Syndrome (SIDS). Despite the importance and strength of the trigeminocardiac reflex, there is little information about the cellular mechanisms and brain stem pathways that constitute this reflex. To address these issues, stimulation of trigeminal afferent fibers and the evoked excitatory postsynaptic currents were recorded in cardiac vagal neurons (CVNs) in an in vitro brain stem slice preparation. This synaptic pathway is robust and activation of the trigeminal pathway often evoked action potentials in CVNs. Application of the serotonin (5-HT) reuptake inhibitor citalopram significantly enhanced these responses. Consistent with the hypothesis this pathway is endogenously modulated by 5-HT receptors the 5-HT1A receptor antagonist, WAY 100635 inhibited, whereas the 5-HT2A/C receptor antagonist, ketanserin facilitated the excitatory neurotransmission to CVNs. The 5-HT1A receptor agonist 8-hydroxy-2-(dipropylamino)tetralin hydrobromide increased, whereas the 5-HT2 receptor agonist, alpha-methylserotonin maleate salt inhibited this reflex pathway. These results indicate stimulation of trigeminal fibers evokes a powerful excitatory and polysynaptic pathway to CVNs, and this pathway is endogenously modulated and differentially enhanced and depressed, by 5-HT1A and 5 HT2 receptors, respectively. PMID- 19553491 TI - Intrinsic membrane properties and inhibitory synaptic input of kenyon cells as mechanisms for sparse coding? AB - The insect mushroom bodies (MBs) are multimodal signal processing centers and are essential for olfactory learning. Electrophysiological recordings from the MBs' principal component neurons, the Kenyon cells (KCs), showed a sparse representation of olfactory signals. It has been proposed that the intrinsic and synaptic properties of the KC circuitry combine to reduce the firing of action potentials and to generate relatively brief windows for synaptic integration in the KCs, thus causing them to operate as coincidence detectors. To better understand the ionic mechanisms that mediate the KC intrinsic firing properties, we used whole cell patch-clamp recordings from KCs in the adult, intact brain of Periplaneta americana to analyze voltage- and/or Ca2+-dependent inward (ICa, INa) and outward currents [IA, IK(V), IK,ST, IO(Ca)]. In general the currents had properties similar to those of currents in other insect neurons. Certain functional parameters of ICa and IO(Ca), however, had unusually high values, allowing them to assist sparse coding. ICa had a low-activation threshold and a very high current density compared with those of ICa in other insect neurons. Together these parameters make ICa suitable for boosting and sharpening the excitatory postsynaptic potentials as reported in previous studies. IO(Ca) also had a large current density and a very depolarized activation threshold. In combination, the large ICa and IO(Ca) are likely to mediate the strong spike frequency adaptation. These intrinsic properties of the KCs are likely to be supported by their tonic, inhibitory synaptic input, which was revealed by specific GABA antagonists and which contributes significantly to the hyperpolarized membrane potential at rest. PMID- 19553493 TI - Direction-dependent control of balance during walking and standing. AB - Human walking has previously been described as "controlled falling." Some computational models, however, suggest that gait may also have self-stabilizing aspects requiring little CNS control. The fore-aft component of walking may even be passively stable from step to step, whereas lateral motion may be unstable and require motor control for balance, as through active foot placement. If this is the case, walking humans might rely less on integrative sensory feedback, such as vision, for anteroposterior (AP) than for mediolateral (ML) balance. We tested whether healthy humans (n=10) exhibit such direction-dependent control, by applying low-frequency perturbations to the visual field (a projected virtual hallway) and measuring foot placement during treadmill walking. We found step variability to be nearly 10 times more sensitive to ML than to AP perturbations, as quantified by the increase in root-mean-square step variability per unit change in perturbation amplitude. This is not simply due to poorer physiological sensitivity of vision in the AP direction: similar perturbations applied to quiet standing produced reversed direction dependence, with an AP sensitivity 2.3-fold greater than that of ML. Tandem (heel-to-toe) standing yielded ML sensitivity threefold greater than that of AP, suggesting that the base of support influences the stability of standing. Postural balance nevertheless appears to require continuous, integrative motor control for balance in all directions. In contrast, walking balance requires step-by-step, integrative control for balance, but mainly in the lateral direction. In the fore-aft direction, balance may be maintained through an "uncontrolled," yet passively stabilized, series of falls. PMID- 19553494 TI - Early and late modulation of saccade deviations by target distractor similarity. AB - In this study, we investigated the time course of oculomotor competition between bottom-up and top-down selection processes using saccade trajectory deviations as a dependent measure. We used a paradigm in which we manipulated saccade latency by offsetting the fixation point at different time points relative to target onset. In experiment 1, observers made a saccade to a filled colored circle while another irrelevant distractor circle was presented. The distractor was either similar (i.e., identical) or dissimilar to the target. Results showed that the strength of saccade deviation was modulated by target distractor similarity for short saccade latencies. To rule out the possibility that the similar distractor affected the saccade trajectory merely because it was identical to the target, the distractor in experiment 2 was a square shape of which only the color was similar or dissimilar to the target. The results showed that deviations for both short and long latencies were modulated by target distractor similarity. When saccade latencies were short, we found less saccade deviation away from a similar than from a dissimilar distractor. When saccade latencies were long, the opposite pattern was found: more saccade deviation away from a similar than from a dissimilar distractor. In contrast to previous findings, our study shows that task-relevant information can already influence the early processes of oculomotor control. We conclude that competition between saccadic goals is subject to two different processes with different time courses: one fast activating process signaling the saliency and task relevance of a location and one slower inhibitory process suppressing that location. PMID- 19553492 TI - Effects of 17beta-estradiol on responses of viscerosomatic convergent thalamic neurons in the ovariectomized female rat. AB - Ovarian hormones have been shown to exert multiple effects on CNS function and viscerosomatic convergent activity. Ovariectomized (OVX) female rats were used in the present study to examine the long-term effects of proestrus levels of 17beta estradiol (EB) delivered by a 60-day time-released subcutaneous pellet on the response properties of viscerosomatic convergent thalamic neurons. In addition, avoidance thresholds to mechanical stimulation for one of the convergent somatic territories, the trunk, was assessed using an electro-von Frey anesthesiometer before and at the end of the 6-wk post-OVX/implant period prior to the terminal electrophysiological experiments, which were done under urethane anesthesia. Rats implanted with an EB-containing pellet, relative to placebo controls, demonstrated 1) altered thalamic response frequencies and thresholds for cervix and vaginal but not colon stimulation; 2) some response variations for just the lateral group of thalamic subnuclei; and 3) altered thalamic response frequencies and thresholds for trunk stimulation. Thalamic response thresholds for trunk pressure in EB versus placebo rats were consistent with the avoidance thresholds obtained from the same groups. In addition, EB replacement affected visceral and somatic thresholds in opposite ways (i.e., reproductive-related structures were less sensitive to pressure, whereas somatic regions showed increased sensitivity). These results have obvious reproductive advantages (i.e., decreased reproductive organ sensitivity for copulation and increased trunk sensitivity for lordosis posturing), as well as possible clinical implications in women suffering from chronic pelvic pain syndromes and/or neuropathic pain. PMID- 19553495 TI - Methazolamide does not impair respiratory work performance in anesthetized rabbits. AB - In human medicine, the carbonic anhydrase (CA) inhibitor acetazolamide is used to treat irregular breathing disorders. Previously, we demonstrated in the rabbit that this substance stabilized closed-loop gain properties of the respiratory control system, but concomitantly weakened respiratory muscles. Among others, the highly diffusible CA-inhibitor methazolamide differs from acetazolamide in that it fails to activate Ca(2+)-dependent potassium channels in skeletal muscles. Therefore, we aimed to find out, whether or not methazolamide may exert attenuating adverse effects on respiratory muscle performance as acetazolamide. In anesthetized spontaneously breathing rabbits (n = 7), we measured simultaneously the CO(2) responses of tidal phrenic nerve activity, tidal transpulmonary pressure changes, and tidal volume before and after intravenous application of methazolamide at two mean (+/- SE) cumulative doses of 3.5 +/- 0.1 and 20.8 +/- 0.4 mg/kg. Similar to acetazolamide, low- and high-dose methazolamide enhanced baseline ventilation by 52 +/- 10% and 166 +/- 30%, respectively (P < 0.01) and lowered the base excess in a dose-dependent manner by up to 8.3 +/- 0.9 mmol/l (P < 0.001). The transmission of a CO(2)-induced rise in phrenic nerve activity into volume and/or pressure and, hence, respiratory work performance was 0.27 +/- 0.05 ml x kg(-1) x kPa x unit(-1) under control conditions, but remained unchanged upon low- or high-dose methazolamide, at 0.30 +/- 0.06 and 0.28 +/- 0.07 ml x kg(-1) x kPa x unit(-1), respectively. We conclude that methazolamide does not cause respiratory muscle weakening at elevated levels of ventilatory drive. This substance (so far not used for medication of respiratory diseases) may thus exert stabilizing influences on breathing control without adverse effects on respiratory muscle function. PMID- 19553496 TI - Olive oil-supplemented diet alleviates acute heat stress-induced mitochondrial ROS production in chicken skeletal muscle. AB - We have previously shown that avian uncoupling protein (avUCP) is downregulated on exposure to acute heat stress, stimulating mitochondrial reactive oxygen species (ROS) production and oxidative damage. In this study, we investigated whether upregulation of avUCP could attenuate oxidative damage caused by acute heat stress. Broiler chickens (Gallus gallus) were fed either a control diet or an olive oil-supplemented diet (6.7%), which has been shown to increase the expression of UCP3 in mammals, for 8 days and then exposed either to heat stress (34 degrees C, 12 h) or kept at a thermoneutral temperature (25 degrees C). Skeletal muscle mitochondrial ROS (measured as H(2)O(2)) production, avUCP expression, oxidative damage, mitochondrial membrane potential, and oxygen consumption were studied. We confirmed that heat stress increased mitochondrial ROS production and malondialdehyde levels and decreased the amount of avUCP. As expected, feeding birds an olive oil-supplemented diet increased the expression of avUCP in skeletal muscle mitochondria and decreased ROS production and oxidative damage. Studies on mitochondrial function showed that heat stress increased membrane potential in state 4, which was reversed by feeding birds an olive oil-supplemented diet, although no differences in basal proton leak were observed between control and heat-stressed groups. These results show that under heat stress, mitochondrial ROS production and olive oil-induced reduction of ROS production may occur due to changes in respiratory chain activity as well as avUCP expression in skeletal muscle mitochondria. PMID- 19553497 TI - Going with the Wnt? Focus on "Hyperaldosteronism, hypervolemia, and increased blood pressure in mice expressing defective APC". PMID- 19553498 TI - Chronic ethanol attenuates circadian photic phase resetting and alters nocturnal activity patterns in the hamster. AB - Acute ethanol (EtOH) administration impairs circadian clock phase resetting, suggesting a mode for the disruptive effect of alcohol abuse on human circadian rhythms. Here, we extend this research by characterizing the chronobiological effects of chronic alcohol consumption. First, daily profiles of EtOH were measured in the suprachiasmatic nucleus (SCN) and subcutaneously using microdialysis in hamsters drinking EtOH. In both cases, EtOH peaked near lights off and declined throughout the dark-phase to low day-time levels. Drinking bouts preceded EtOH peaks by approximately 20 min. Second, hamsters chronically drinking EtOH received a light pulse during the late dark phase [Zeitgeber time (ZT) 18.5] to induce photic phase advances. Water controls had shifts of 1.2 +/- 0.2 h, whereas those drinking 10% and 20% EtOH had much reduced shifts (0.5 +/- 0.1 and 0.3 +/- 0.1 h, respectively; P < 0.001 vs. controls). Third, incremental decreases in light intensity (270 lux to 0.5 lux) were used to explore chronic EtOH effects on photic entrainment and rhythm stability. Activity onset was unaffected by 20% EtOH at all light intensities. Conversely, the 24-h pattern of activity bouts was disrupted by EtOH under all light intensities. Finally, replacement of chronic EtOH with water was used to examine withdrawal effects. Water controls had photic phase advances of 1.1 +/- 0.3 h, while hamsters deprived of EtOH for 2-3 days showed enhanced shifts (2.1 +/- 0.3 h; P < 0.05 vs. controls). Thus, in chronically drinking hamsters, brain EtOH levels are sufficient to inhibit photic phase resetting and disrupt circadian activity. Chronic EtOH did not impair photic entrainment; however, its replacement with water potentiated photic phase resetting. PMID- 19553500 TI - Comparative effects of oral and intraduodenal glucose on blood pressure, heart rate, and splanchnic blood flow in healthy older subjects. AB - Postprandial hypotension occurs frequently, particularly in the elderly. The magnitude of the fall in blood pressure (BP) and rise in heart rate (HR) in response to enteral glucose are greater when gastric emptying (GE) or small intestinal infusion are more rapid. Meal ingestion is associated with an increase in splanchnic blood flow. In contrast, gastric distension may attenuate the postprandial fall in BP. The aims of this study were to evaluate, in older subjects, the comparative effects of intraduodenal glucose infusion, at a rate similar to GE of oral glucose, on BP, HR, superior mesenteric artery (SMA) flow, and blood glucose. Eight healthy subjects (5 men, 3 women, age 66-75 yr) were studied on two occasions. On day 1, each subject ingested 300 ml of water containing 75 g glucose. GE was quantified by three-dimensional ultrasonography between time t = 0-120 min, and the rate of emptying (kcal/min) was calculated. On day 2, glucose was infused intraduodenally at the same rate as that on day 1. On both days, BP, HR, SMA flow, and blood glucose were measured. The mean GE of oral glucose was 1.3 +/- 0.1 kcal/min. Systolic BP (P < 0.01), SMA flow (P < 0.05), and blood glucose (P < 0.01) were greater and HR less (P < 0.01) after oral, compared with intraduodenal, glucose. There were comparable falls in diastolic BP during the study days (P < 0.01 for both). We conclude that the magnitude of the fall in systolic BP and rise in HR are less after oral, compared with intraduodenal, glucose, presumably reflecting the "protective" effect of gastric distension. PMID- 19553499 TI - Functional overload in ground squirrel plantaris muscle fails to induce myosin isoform shifts. AB - We performed 2 wk of mechanical overload by synergist ablation on plantaris muscles from a small rodent hibernator, Spermophilus lateralis. While this muscle displays prominent myosin heavy-chain (MyHC) isoform shifts during hibernation, sensitivity to mechanical loading as a stimulus for muscle mass and isoform plasticity has not been demonstrated. Squirrel muscles, whether during hibernation or not, potentially are less sensitive to mechanical unloading, but we hypothesized that increased loading would produce the typical mammalian response of greater plantaris mass and MyHC shifts. Mechanical overload produced a 50% increase in muscle mass but, surprisingly, no changes in MyHC isoform protein or mRNA expression, despite previously observed fast-to-slow MyHC isoform switching during hibernation. Citrate synthase enzyme activity, as well as mRNA expression of creatine kinase and the muscle growth factor myostatin, were all unchanged. The mRNA expression of critical muscle atrophy genes decreased by 50% during hypertrophy, including ubiquitin ligases MuRF1 and MAFbx, and the related transcription factor FOXO-1a. Insulin-like growth factor (IGF-1) and hypoxia inducible factor (HIF-1alpha) mRNA expression was elevated by 400% and 150%. Fast to-slow MyHC isoform shifts appear unnecessary to support the increased recruitment of the plantaris muscle, shifts which are seen in other rodent models. Our results are consistent with muscular activity during interbout arousals as a potential mechanism to preserve muscle mass, but illustrate the primary importance of other seasonal factors besides patterns of muscle activation which must act in concert to alter MyHC isoforms and muscle fiber type during hibernation. PMID- 19553501 TI - Pressure-induced constriction is inhibited in a mouse model of reduced betaENaC. AB - Recent studies suggest certain epithelial Na(+) channel (ENaC) proteins may be components of mechanosensitive ion channel complexes in vascular smooth muscle cells that contribute to pressure-induced constriction in middle cerebral arteries (MCA). However, the role of a specific ENaC protein, betaENaC, in pressure-induced constriction of MCAs has not been determined. The goal of this study was to determine whether pressure-induced constriction in the MCA is altered in a mouse model with reduced levels of betaENaC. Using quantitative immunofluorescence, we found whole cell betaENaC labeling in cerebral vascular smooth muscle cells (VSMCs) was suppressed 46% in betaENaC homozygous mutant (m/m) mice compared with wild-type littermates (+/+). MCAs from betaENaC +/+ and m/m mice were isolated and placed in a vessel chamber for myographic analysis. Arteries from betaENaC+/+ mice constricted to stepwise increases in perfusion pressure and developed maximal tone of 10 +/- 2% at 90 mmHg (n = 5). In contrast, MCAs from betaENaC m/m mice developed significantly less tone (4 +/- 1% at 90 mmHg, n = 5). Vasoconstrictor responses to KCl (4-80 mM) were identical between genotypes and responses to phenylephrine (10(-7)-10(-4) M) were marginally altered, suggesting that reduced levels of VSMC betaENaC specifically inhibit pressure-induced constriction. Our findings indicate betaENaC is required for normal pressure-induced constriction in the MCA and provide further support for the hypothesis that betaENaC proteins are components of a mechanosensor in VSMCs. PMID- 19553502 TI - Neuronostatin inhibits cardiac contractile function via a protein kinase A- and JNK-dependent mechanism in murine hearts. AB - Neuronostatin, a newly identified peptide hormone sharing the same precursor with somatostatin, exerts multiple pharmacological effects in gastrointestinal tract, hypothalamus, and cerebellum. However, the cardiovascular effect of neuronostatin is unknown. The aim of this study was to elucidate the impact of neuronostatin on cardiac contractile function in murine hearts and isolated cardiomyocytes. Short term exposure of neuronostatin depressed left ventricular developed pressure (LVDP), maximal velocity of pressure development (+/-dP/dt), and heart rate in Langendorff heart preparation. Consistently, neuronostatin inhibited peak shortening (PS) and maximal velocity of shortening/relengthening (+/-dL/dt) without affecting time-to-PS (TPS) and time-to-90% relengthening (TR(90)) in cardiomyocytes. The neuronostatin-elicited cardiomyocyte mechanical responses were mimicked by somatostatin, the other posttranslational product of preprosomatostatin. Furthermore, the neuronostatin-induced cardiomyocyte mechanical effects were ablated by the PKA inhibitor H89 (1 microM) and the Jun N terminal kinase (JNK) inhibitor SP600125 (20 microM). The PKC inhibitor chelerythrine (1 microM) failed to alter neuronostatin-induced cardiomyocyte mechanical responses. To the contrary, chelerythrine, but not H89, abrogated somatostatin-induced cardiomyocyte contractile responses. Our results also showed enhanced c-fos and c-jun expression in response to neuronostatin exposure (0.5 to 2 h). Taken together, our data suggest that neuronostatin is a peptide hormone with overt cardiac depressant action. The neuronostatin-elicited cardiac contractile response appears to be mediated, at least in part, through a PKA- and/or JNK-dependent mechanism. PMID- 19553503 TI - Metformin improves postprandial glucose homeostasis in rainbow trout fed dietary carbohydrates: a link with the induction of hepatic lipogenic capacities? AB - Carnivorous fish are poor users of dietary carbohydrates and are considered to be glucose intolerant. In this context, we have tested, for the first time in rainbow trout, metformin, a common anti-diabetic drug, known to modify muscle and liver metabolism and to control hyperglycemia in mammals. In the present study, juvenile trout were fed with very high levels of carbohydrates (30% of the diet) for this species during 10 days followed by feeding with pellets supplemented with metformin (0.25% of the diet) for three additional days. Dietary metformin led to a significant reduction in postprandial glycemia in trout, demonstrating unambiguously the hypoglycemic effect of this drug. No effect of metformin was detected on mRNA levels for glucose transporter type 4 (GLUT4), or enzymes involved in glycolysis, mitochondrial energy metabolism, or on glycogen level in the white muscle. Expected inhibition of hepatic gluconeogenic (glucose-6 phosphatase, fructose-1,6-bisphosphatase, and phosphoenolpyruvate carboxykinase) mRNA levels was not found, showing instead paradoxically higher mRNA levels for these genes after drug treatment. Finally, metformin treatment was associated with higher mRNA levels and activities for lipogenic enzymes (fatty acid synthase and glucose-6-phosphate dehydrogenase). Overall, this study strongly supports that the induction of hepatic lipogenesis by dietary glucose may permit a more efficient control of postprandial glycemia in carnivorous fish fed with high carbohydrate diets. PMID- 19553504 TI - Electrophysiological responses of sympathetic preganglionic neurons to ANG II and aldosterone. AB - The intermediolateral cell column (IML) of the spinal cord is an important area where sympathetic impulses propagate to peripheral sympathetic organs. ANG II and aldosterone are important components of the renin-angiotensin-aldosterone system (RAAS), which activate the sympathetic nervous system. Each is partly synthesized in the brain and plays a paracrine role in the regulation of blood pressure independently of RAAS in the periphery. Our purpose in the present study was to clarify the contributions of sympathetic preganglionic neurons in the IML (IML neurons) and the effects of ANG II and aldosterone on the sympathetic nervous system. To examine responses to ANG II and aldosterone, we intracellularly recorded 104 IML neurons using a whole cell patch-clamp technique in spinal cord slice preparations. IML neurons were classified into two types: silent and firing. Both neuron types were significantly depolarized by ANG II, and candesartan inhibited this depolarization. After pretreatment with TTX, firing neurons (but not silent neurons) were significantly depolarized by ANG II. Aldosterone significantly increased the number of excitatory postsynaptic potentials (EPSPs) in both neuron types, but this response disappeared after pretreatment with TTX. ANG II and aldosterone had no synergistic effects on the IML neurons. The silent neurons had large cell soma, and many more dendrites than the firing neurons. These results suggest that ANG II acts presynaptically and postsynaptically in IML neurons, while aldosterone acts mainly presynaptically. Thus, the physiological effects of these substances are likely to be transmitted via specific membrane receptors of IML and/or presynaptic neurons. PMID- 19553505 TI - Mutations in influenza virus replication and transcription: detection of amino acid substitutions in hemagglutinin of an avian influenza virus (H1N1). AB - Influenza A viruses are RNA viruses that contain negative-sense, single-stranded, and segmented RNA genome, which depends on virally encoded RNA-dependent RNA polymerase and cellular DNA-dependent RNA polymerase for replication of viral genome and transcription of viral mRNA, respectively. Hemagglutinin (HA), one of the major surface proteins of the influenza virus, is responsible for virus attachment to the receptor of host cells to initiate an infection. Amino acid (AA) substitutions in HA may cause changes in virus antigenicity and even receptor specificity. To detect the AA substitutions within HA at protein level, nanoelectrospray-MS/MS was used to analyze tryptic digestion of HA antigen directly purified from virus particles of an avian influenza virus, A/WDK/JX/12416/2005 (H1N1), of which the HA gene was sequenced as a reference. The comparison of the sequences obtained from analysis of viral genome and peptide found seven variations between HA gene and protein, namely E103K, R130K, T169I, I338V, N387S, S398I/L, and I399S in HA. Because influenza virus uses different polymerase machineries for replication and transcription, these substitutions could be introduced in the viral genome through replication process but not in viral mRNA in the transcription. The results, for the first time, provided experimental evidence showing differences in AA sequence obtained from direct analysis of viral protein derived from viral genome. PMID- 19553507 TI - Chasing the suspect: keratoconus. PMID- 19553508 TI - Carving the cornea: the von Hippel trephine. PMID- 19553509 TI - Emerging options in the management of advanced intraocular retinoblastoma. PMID- 19553510 TI - In vivo confocal microscopy of the ocular surface: where are we now? PMID- 19553511 TI - Femtosecond dovetail penetrating keratoplasty: surgical technique and case report. AB - AIM: To report a dovetail configuration for femtosecond-enabled penetrating keratoplasty (PK) with the corresponding laser parameters and suturing technique. METHODS: A 40-year-old man, with a history of penetrating corneal injury as a child, underwent femtosecond-enabled dovetail keratoplasty, anterior vitrectomy and secondary intraocular lens suturing to repair his corneal scar and aphakia. A partial thickness dovetail pattern was performed in the recipient cornea using the femotsecond laser. The posterior side-cut was initiated approximately 100 microm anterior to the Descemet membrane and extended obliquely towards the outer edge of a ring lamellar cut, positioned at approximately 300 microm stromal depth. The anterior side-cut was extended from the internal edge of the ring lamellar cut to the corneal surface. Using an artificial chamber, the femtosecond laser was used to create a full-thickness 0.2 mm oversized femtosecond-enabled dovetail trephination with similar anterior lamellar depth (approximately 300 microm). Wound closure, using interrupted 10-0 nylon sutures, was guided by preplaced radial alignment laser microincisions and tongue-in-groove midstromal suture positioning. RESULTS AND DISCUSSION: Excellent alignment and stability of the donor and recipient tissue were observed immediately postoperatively and 5 months after surgery. The feasibility of the "dovetail" pattern of PK and the tongue-in-groove suture positioning is demonstrated. PMID- 19553512 TI - Treatment of inflamed pterygia or residual pterygial bed. AB - AIMS: To describe the use of subconjunctival bevacizumab or ranibizumab, an approved antivascular endothelial growth factor for wet macular degeneration, in halting the inflammation of a pterygium or a partially excised pterygium. METHODS: Case reports. RESULTS: Prompt regression of conjunctival microvessels in the pterygial bed was documented 1 week after a single subconjunctival injection of ranibizumab (one case) or bevacizumab (two cases). No side-effects were noted over 13 months of follow-up in the first case, 6 months in the second case and 1 month in the third case. CONCLUSION: Selective blockade of vascular endothelial growth factor was effective in causing regression of conjunctival microvessels in three eyes with inflamed pterygium or residual pterygia. PMID- 19553514 TI - Rituximab for treatment of scleritis associated with rheumatoid arthritis. PMID- 19553515 TI - Film intraocular lens: a new way to choose the light-transmission spectrum. PMID- 19553517 TI - Cataract surgery and mortality. PMID- 19553518 TI - Promise and potential pitfalls of anti-VEGF drugs in retinopathy of prematurity. PMID- 19553519 TI - Heterodisaccharide 4-O-(N-acetyl-beta-D-glucosaminyl)-D-glucosamine is a specific inducer of chitinolytic enzyme production in Vibrios harboring chitin oligosaccharide deacetylase genes. AB - Vibrio parahaemolyticus KN1699 produces 4-O-(N-acetyl-beta-d-glucosaminyl)-d glucosamine (GlcNAc-GlcN) as a major end product from chitin using two extracellular hydrolases: glycoside hydrolase family 18 chitinase, which produces (GlcNAc)(2) from chitin, and carbohydrate esterase (CE) family 4 chitin oligosaccharide deacetylase (COD), which hydrolyzes the N-acetyl group at the reducing-end GlcNAc residue of (GlcNAc)(2). In this study, we clarified that this heterodisaccharide functions as an inducer of the production of the two above mentioned chitinolytic enzymes, particularly chitinase. Similar results for chitinase production were obtained with other chitin-decomposing Vibrio strains harboring the CE family 4 COD gene; however, such an increase in chitinase production was not observed in chitinolytic Vibrio strains that did not harbor the COD gene. These results suggest that GlcNAc-GlcN is a unique inducer of chitinase production in Vibrio bacteria that have the COD-producing ability and that the COD involved in the synthesis of this signal compound is one of the key enzymes in the chitin catabolic cascade of these bacteria. PMID- 19553520 TI - A20 attenuates allergic airway inflammation in mice. AB - TNF receptor 1 can activate signaling pathways leading to the activation of NF kappaB. A20, an NF-kappaB-inducible protein, negatively regulates these signaling pathways and acts as an anti-inflammatory mediator. Therefore, A20 is viewed as a potential therapeutic target for inflammatory disease. In this study, we examined the effect of A20 on an OVA-induced allergic airway inflammation model in mice. We used an adenovirus containing A20 cDNA (Ad-A20) that was delivered intratracheally before OVA challenge. Single administration of Ad-A20 reduced airway inflammatory cell recruitment and peribronchiolar inflammation and suppressed the production of various cytokines in bronchoalveolar fluid. In addition, Ad-A20 suppressed mucus production and prevented the development of airway hyperresponsiveness. The protective effect of Ad-A20 was mediated by the inhibition of the NF-kappaB signaling pathway. Taken together, our results suggest that the development of an immunoregulatory strategy based on A20 may have therapeutic potential for the treatment of allergic asthma. PMID- 19553523 TI - Initiation of antigen receptor-dependent differentiation into plasma cells by calmodulin inhibition of E2A. AB - Differentiation of B lymphocytes into Ab-secreting plasmablasts and plasma cells is Ag driven. The interaction of Ag with the membrane-bound Ab of the BCR is critical in determining which clones enter the plasma cell response. However, not much is known about the coupling between BCR activation and the shift in transcription factor network from that of a B cell to that of ASC differentiation. Our genome-wide analysis shows that Ab-secreting cell differentiation of mouse B cells is induced by BCR activation through very fast regulatory events from the BCR. We identify activation of IFN regulatory factor-4 and down-regulation of Pax5, Bcl-6, MITF, Ets-1, Fli-1, and Spi-B gene expression as immediate early events. Furthermore, the transcription factor E2A is required for the rapid key down-regulations after BCR activation, and the Ca(2+) sensor protein calmodulin has the corresponding regulatory effect as BCR activation. Moreover, mutants in the calmodulin binding site of E2A show that Ca(2+) signaling through calmodulin inhibition of E2A is essential for the rapid down regulation of immediate early genes after BCR activation in initiation of plasma cell differentiation. PMID- 19553522 TI - The polarity protein Par1b/EMK/MARK2 regulates T cell receptor-induced microtubule-organizing center polarization. AB - Engagement of a T cell to an APC induces the formation of an immunological synapse as well as reorientation of the microtubule-organizing center (MTOC) toward the APC. How signals emanating from the TCR induce MTOC polarization is not known. One group of proteins known to play a critical role in asymmetric cell division and cell polarization is the partitioning defective (Par) family of proteins. In this study we found that Par1b, a member of the Par family of proteins, was inducibly phosphorylated following TCR stimulation. This phosphorylation resulted in 14-3-3 protein binding and caused the relocalization of Par1b from the membrane into the cytoplasm. Because a dominant-negative form of Par1b blocked TCR-induced MTOC polarization, our data suggest that Par1b functions in the establishment of T cell polarity following engagement to an APC. PMID- 19553521 TI - IL-2 regulates CD103 expression on CD4+ T cells in Scurfy mice that display both CD103-dependent and independent inflammation. AB - Scurfy (Sf) mice lack CD4(+)Foxp3(+) regulatory T cells and develop fatal multiorgan inflammation (MOI) mediated by CD4(+) T cells. Introducing Il2(-/-) gene into Sf mice (Sf.Il2(-/-)) inhibited inflammation in skin and lung. As a major integrin receptor for the organs, we compared CD103 expression on the CD4(+) T cells of B6, Il2(-/-), Sf, and Sf.Il2(-/-) mice. CD103(+)CD4(+) T cells, but not CD8(+) T cells or CD11c(+) dendritic cells, were significantly up regulated only in Sf mice, indicating Il2(-/-) dominantly and specifically inhibited CD103 up-regulation in Sf CD4(+) T cells. In addition, CD4(+)Foxp3(+) regulatory T cell CD103 expression was not reduced in Il2(-/-) mice. Introducing CD103(-/-) into Sf mice inhibited inflammation in skin and lung as compared with age-matched Sf mice, but they died at approximately 7 wk old with inflammation developed in skin, lungs, and colon, demonstrating fatal MOI induced by CD103 independent mechanism. Transfer of Sf CD4(+) T cells induced MOI more rapidly than CD103(-)CD4(+) T cells, indicating the presence of CD103-dependent mechanism for inflammation. In vitro stimulation with anti-CD3 plus anti-CD28 beads confirmed that CD103 induction in the CD4(+)Foxp3(-) T cells in Il2(-/-) and Sf.Il2(-/-) is defective and cannot be restored by rIL-2 or rIL-15. The data indicate that IL-2 is required for optimal CD103 induction on CD4(+) T cells in Sf mice and this effect contributes to inflammation in an organ-specific manner. IL-2 also has additional roles because the protection of skin and lung inflammation in Sf.Il2(-/-), but not Sf.CD103(-/-) mice is lifelong and Sf.Il2(-/ ) mice have longer lifespan than Sf.CD103(-/-) mice. PMID- 19553524 TI - Expression of a modified form of CD4 results in the release of an anti-HIV factor derived from the Env sequence. AB - We have studied the inhibitory effect of a CD4 chimera (CD4epsilon15) on HIV replication. This chimera is retained in the endoplasmic reticulum and traps the HIV envelope precursor gp160, preventing its maturation. Retroviral expression of the chimera strongly inhibited HIV replication even when it is expressed by only a minority of the T cell population. This protective effect on bystander nontransduced cells is mediated by a soluble factor that we identified as a fragment of HIV gp120 envelope protein and accordingly, we named this factor Env derived antiviral factor (EDAF). Biochemical and immunoreactivity data show that EDAF is comprised of the gp120 C3-C5 regions and indeed, a recombinant protein bearing this sequence reproduces the anti-HIV properties of EDAF. Surprisingly, three tryptic peptides derived from EDAF are homologous but not identical with the corresponding sequences of the HIV isolate used to generate EDAF. We propose that EDAF results from an alternative intracellular processing of the Env protein provoked by its association to CD4epsilon15 and the selection of the best fitted Env protein sequences contained within the HIV isolate. The presence of EDAF improves the therapeutic potential of the CD4epsilon15 gene and it opens new possibilities for antiviral treatment and vaccine development. PMID- 19553525 TI - Progesterone inhibits activation-induced deaminase by binding to the promoter. AB - Regulation of activation-induced deaminase (AID), an essential factor in Ig diversification, can alter not only somatic hypermutation and class switch recombination (CSR), but may also influence oncogenesis. AID deaminates cytosine to uracil in the Ig locus, thereby initiating Ig diversification. Unregulated AID can induce oncogenic DNA alterations in Ig and non-Ig loci, leading to mutations, recombination, and translocations. In this study, we demonstrate that AID mRNA production in activated mouse splenic B cells can be reduced by treatment with the sex hormone progesterone. This down-regulation is independent of translation or splicing and is predominantly achieved by inhibiting transcription. During cell treatment we could detect progesterone receptor bound to the AID promoter in proximity to NF-kappaB binding. Importantly, the progesterone-induced repression was also extended to the protein level of AID and its activity on somatic hypermutation and class switch recombination. PMID- 19553526 TI - Menopause and ovariectomy cause a low grade of systemic inflammation that may be prevented by chronic treatment with low doses of estrogen or losartan. AB - The incidence of cardiovascular diseases in premenopausal women is lower than in men or postmenopausal women. This study reports the discovery of a low grade of systemic inflammation, including monocyte adhesion to arterial endothelium, elicited by menopause or estrogen depletion. Chronic treatment with low dose of 17-beta-estradiol or inhibition of the renin-angiotensin system reduced this inflammation. Using an in vitro flow chamber system with human arterial and venous endothelial cells, we found that leukocytes from healthy postmenopausal women were more adhesive to the arterial endothelium than those from premenopausal women regardless of the stimulus used on endothelial cells. Increased circulating levels of IL-8, MCP-1, RANTES, and MIP-1alpha and monocyte CD11b expression were also encountered in postmenopausal vs premenopausal subjects. This translational data led us to investigate the mechanisms in Sprague Dawley rats. Using intravital microscopy, we imaged mesenteric arterioles and found significant increases in arteriolar leukocyte adhesion, cell adhesion molecule expression, and plasma levels of cytokine-induced neutrophil chemoattractant (CINC/KC), MCP-1, and MIP-1alpha in 1-mo ovariectomized rats. Chronic treatment of ovariectomized rats with low dose of 17-beta-estradiol, losartan, both, or benazepril inhibited ovariectomy-induced arteriolar mononuclear leukocyte adhesion by 77%, 58%, 92%, and 65% respectively, partly by inhibition of cell adhesion molecule up-regulation and the increase in circulating chemokines. These results demonstrate that menopause and ovariectomy generate a low grade of systemic inflammation. Therefore, administration of low doses of estrogens or inhibition of the renin-angiotensin system, at early stages of estrogen deficiency, might prevent the systemic inflammation associated with menopause and decrease the risk of suffering further cardiovascular diseases. PMID- 19553527 TI - IL-17 and IFN-gamma mediate the elicitation of contact hypersensitivity responses by different mechanisms and both are required for optimal responses. AB - Hapten-induced contact hypersensitivity (CHS) in the skin is a delayed type cellular immune response that can be mediated by CD8(+) T cells that produce IFN gamma or IL-17. However, mechanisms for these cytokines in the elicitation of CHS remain to be fully elucidated. In this study, we show that adoptive transfer of CHS with hapten-primed wild-type (WT) CD8(+) T cells is reduced in IFN-gammaR(-/ ) or IL-17R(-/-) mice compared with WT controls. The infiltration of granulocytes and macrophages in the hapten challenged skin of IL-17R(-/-) recipients is significantly reduced whereas it is less affected in IFN-gammaR(-/-) recipients although CD8(+) T cell infiltration is inhibited in both recipients. In contrast, the activity of reactive oxidative species is significantly inhibited in IFN gammaR(-/-) but is less affected in IL-17R(-/-) recipients. Further analysis reveals that the expression of chemokines and cytokines is differentially regulated in the hapten-challenged skin of IFN-gammaR(-/-) or IL-17R(-/-) recipients compared with WT controls. Interestingly, injection of rIL-17 in the skin induces inflammation with a high level of leukocyte infiltration whereas injection of IFN-gamma induces inflammation with a high level of reactive oxidative species. Moreover, neutralization of IL-17 in IFN-gammaR(-/-) or IFN gamma in IL-17R(-/-) mice further suppresses the adoptive transfer of CHS by hapten-primed WT CD8(+) T cells. The study demonstrates that IFN-gamma and IL-17 mediate the elicitation of CHS by different mechanisms and that both cytokines are required for optimal responses. This outcome improves understanding of pathogenesis and provides new insights into therapeutic strategies for CHS. PMID- 19553529 TI - TLR2 is a negative regulator of Th17 cells and tissue pathology in a pulmonary model of fungal infection. AB - To study the role of TLR2 in a experimental model of chronic pulmonary infection, TLR2-deficient and wild-type mice were intratracheally infected with Paracoccidioides brasiliensis, a primary fungal pathogen. Compared with control, TLR2(-/-) mice developed a less severe pulmonary infection and decreased NO synthesis. Equivalent results were detected with in vitro-infected macrophages. Unexpectedly, despite the differences in fungal loads both mouse strains showed equivalent survival times and severe pulmonary inflammatory reactions. Studies on lung-infiltrating leukocytes of TLR2(-/-) mice demonstrated an increased presence of polymorphonuclear neutrophils that control fungal loads but were associated with diminished numbers of activated CD4(+) and CD8(+) T lymphocytes. TLR2 deficiency leads to minor differences in the levels of pulmonary type 1 and type 2 cytokines, but results in increased production of KC, a CXC chemokine involved in neutrophils chemotaxis, as well as TGF-beta, IL-6, IL-23, and IL-17 skewing T cell immunity to a Th17 pattern. In addition, the preferential Th17 immunity of TLR2(-/-) mice was associated with impaired expansion of regulatory CD4(+)CD25(+)FoxP3(+) T cells. This is the first study to show that TLR2 activation controls innate and adaptive immunity to P. brasiliensis infection. TLR2 deficiency results in increased Th17 immunity associated with diminished expansion of regulatory T cells and increased lung pathology due to unrestrained inflammatory reactions. PMID- 19553528 TI - Trans-presentation of IL-15 by intestinal epithelial cells drives development of CD8alphaalpha IELs. AB - IL-15 is crucial for the development of intestinal intraepithelial lymphocytes (IEL) and delivery is mediated by a unique mechanism known as trans-presentation. Parenchymal cells have a major role in the trans-presentation of IL-15 to IELs, but the specific identity of this cell type is unknown. To investigate whether the intestinal epithelial cells (IEC) are the parenchymal cell type involved, a mouse model that expresses IL-15Ralpha exclusively by the IECs (Villin/IL 15Ralpha Tg) was generated. Exclusive expression of IL-15Ralpha by the IECs restored all the deficiencies in the CD8alphaalpha(+)TCRalphabeta(+)and CD8alphaalpha(+)TCRgammadelta(+) subsets that exist in the absence of IL 15Ralpha. Interestingly, most of the IEL recovery was due to the preferential increase in Thy1(low) IELs, which compose a majority of the IEL population. The differentiation of Thy1(high)CD4(-)CD8(-) thymocytes into Thy1(-)CD8alphaalpha IELs was found to require IL-15Ralpha expression specifically by IECs and thus, provides evidence that differentiation of Thy1(low) IELs is one function of trans presentation of IL-15 in the intestines. In addition to effects in IEL differentiation, trans-presentation of IL-15 by IECs also resulted in an increase in IEL numbers that was accompanied by increases in Bcl-2, but not proliferation. Collectively, this study demonstrates that trans-presentation of IL-15 by IECs alone is completely sufficient to direct the IL-15-mediated development of CD8alphaalpha(+) T cell populations within the IEL compartment, which now includes a newly identified role of IL-15 in the differentiation of Thy1(low) IELs. PMID- 19553530 TI - A subcytotoxic dose of subtilase cytotoxin prevents lipopolysaccharide-induced inflammatory responses, depending on its capacity to induce the unfolded protein response. AB - Subtilase cytotoxin (SubAB) is the prototype of a newly identified family of AB(5) cytotoxins produced by Shiga toxigenic Escherichia coli. SubAB specifically cleaves the essential endoplasmic reticulum (ER) chaperone BiP (GRP78), resulting in the activation of ER stress-induced unfolded protein response (UPR). We have recently shown that the UPR following ER stress can suppress cellular responses to inflammatory stimuli during the later phase, in association with inhibition of NF-kappaB activation. These findings prompted us to hypothesize that SubAB, as a selective UPR inducer, might have beneficial effects on inflammation-associated pathology via a UPR-dependent inhibition of NF-kappaB activation. The pretreatment of a mouse macrophage cell line, RAW264.7, with a subcytotoxic dose of SubAB-triggered UPR and inhibited LPS-induced MCP-1 and TNF-alpha production associated with inhibition of NF-kappaB activation. SubA(A272)B, a SubAB active site mutant that cannot induce UPR, did not show such effects. In addition, pretreatment with a sublethal dose of SubAB, but not SubA(A272)B, protected the mice from LPS-induced endotoxic lethality associated with reduced serum MCP-1 and TNF-alpha levels and also prevented the development of experimental arthritis induced by LPS in mice. Collectively, although SubAB has been identified originally as a toxin associated with the pathogenesis of hemolytic uremic syndrome, the unique ability of SubAB to selectively induce the UPR may have the potential to prevent LPS-associated inflammatory pathology under subcytotoxic conditions. PMID- 19553531 TI - An anti-inflammatory role for plasmacytoid dendritic cells in allergic airway inflammation. AB - It was previously shown that administration of recombinant human Fms-like tyrosine kinase receptor-3 ligand (Flt3L) before allergen challenge of sensitized mice suppresses the cardinal features of asthma through unclear mechanisms. Here, we show that Flt3L dramatically alters the balance of conventional to plasmacytoid dendritic cells (pDCs) in the lung favoring the accumulation of pDCs. Selective removal of pDCs abolished the antiinflammatory effect of Flt3L, suggesting a regulatory role for these cells in ongoing asthmatic inflammation. In support, we found that immature pDCs are recruited to the lungs of allergen challenged mice irrespective of Flt3L treatment. Selective removal of pDCs during allergen challenge enhanced airway inflammation, whereas adoptive transfer of cultured pDCs before allergen challenge suppressed inflammation. Experiments in which TLR9 agonist CpG motifs were administered in vitro or in vivo demonstrated that pDCs were antiinflammatory irrespective of their maturation state. These effects were mediated through programmed death-1/programmed death ligand 1 interactions, but not through ICOS ligand, IDO, or IFN-alpha. These findings suggest a specialized immunoregulatory role for pDCs in airway inflammation. Enhancing the antiinflammatory properties of pDCs could be employed as a novel strategy in asthma treatment. PMID- 19553532 TI - MyD88 and Type I interferon receptor-mediated chemokine induction and monocyte recruitment during Listeria monocytogenes infection. AB - Monocytes play a central role in defense against infection, but the mechanisms promoting monocyte recruitment and activation remain incompletely defined. Defense against Listeria monocytogenes, an intracellular bacterial pathogen, requires in vivo MCP-1 induction and CCR2-dependent recruitment of Ly6C(high) monocytes from bone marrow to sites of infection. Herein, we demonstrate that infection of bone marrow-derived macrophages with virulent L. monocytogenes induces MCP-1 expression in two phases. The first phase is rapid, induces low level production of MCP-1, and is dependent on TLR/MyD88 signaling. The second phase promotes prolonged, higher level MCP-1 secretion and is dependent on signaling via the type I IFN receptor (IFNAR). Although attenuated L. monocytogenes strains that remain confined to the phagosome trigger TLR/MyD88 mediated signals and induce low-level MCP-1 expression, only cytosol-invasive bacteria promote IFNAR-dependent MCP-1 expression. In vivo, deficiency of either MyD88 or IFNAR signaling does not impair early monocyte emigration from bone marrow and recruitment to infected spleen. Loss of both MyD88 and IFNAR-mediated MCP-1 induction, however, results in deficient Ly6C(high) monocyte recruitment and increased susceptibility to L. monocytogenes infection. Our studies demonstrate that distinct but partially overlapping signal transduction pathways provide redundancy that ensures optimal monocyte recruitment to sites of microbial infection. PMID- 19553534 TI - Zymosan, but not lipopolysaccharide, triggers severe and progressive peritoneal injury accompanied by complement activation in a rat peritonitis model. AB - Fungal peritonitis is an important complication in peritoneal dialysis patients; either continuous or recurrent peritonitis may enhance peritoneal damage. Even when the peritoneal dialysis catheter is removed in patients with fungal peritonitis, peritoneal fibrosis can progress and evolve into encapsular peritoneal sclerosis. It is unclear why fungal infections are worse than bacterial in these respects. Zymosan is a cell wall component of yeast that strongly activates the complement system. In this study, we compared the effects of zymosan and bacterial LPS on peritoneal inflammation in a rat peritoneal injury model induced by mechanical scraping. Intraperitoneal administration of zymosan, but not LPS or vehicle, caused markedly enhanced peritonitis with massive infiltration of cells and deposition of complement activation products C3b and membrane attack complex on day 5. In rats administered zymosan and sacrificed on days 18 or 36, peritoneal inflammation persisted with accumulation of ED-1-positive cells, small deposits of C3b and membrane attack complex, exudation of fibrinogen, and capillary proliferation in subperitoneal tissues. When zymosan was administered daily for 5 days after peritoneal scrape, there was even greater peritoneal inflammation with peritoneal thickening, inflammatory cell accumulation, and complement deposition. Inhibition of systemic complement by pretreatment with cobra venom factor or local inhibition by i.p. administration of the recombinant complement regulator Crry-Ig reduced peritoneal inflammation in zymosan-treated rats. Our results show that yeast components augment inflammation in the injured peritoneum by causing complement activation within the peritoneal cavity. Local anticomplement therapy may therefore protect from peritoneal damage during fungal infection of the peritoneum. PMID- 19553535 TI - Ncf1-associated reduced oxidative burst promotes IL-33R+ T cell-mediated adjuvant free arthritis in mice. AB - Reactive oxygen species (ROS) are important in the immune defense against invading pathogens, but they are also key molecules in the regulation of inflammatory reactions. Low levels of ROS production due to a polymorphism in the neutrophil cytosolic factor 1 (Ncf1) gene are associated with autoimmunity and arthritis severity in mouse models induced with adjuvant. We established an adjuvant-free arthritis model in which disease is induced by injection of the autoantigen collagen type II (CII) and depends on IL-5-producing T cells and eosinophils. In addition, the transgenic expression of mutated mouse CII allowed us to investigate an autoreactive immune response to an autologous Ag and by that natural tolerance mechanism. We show that a deficient ROS production, due to a spontaneous mutation in Ncf1, leads to increased autoantibody production and expansion of IL-33R-expressing T cells, impaired T cell tolerance toward tissue specific CII, and severe arthritis in this unique model without disturbing adjuvant effects. These results demonstrate that the insufficient production of ROS promotes the breakdown of immune tolerance and development of autoimmune and adjuvant-free arthritis through an IL-5- and IL33R-dependent T cell activation pathway. PMID- 19553533 TI - Myeloid-derived suppressor cells down-regulate L-selectin expression on CD4+ and CD8+ T cells. AB - Effective cell-mediated antitumor immunity requires the activation of tumor reactive T cells and the trafficking of activated T cells to tumor sites. These processes involve the extravasation of lymphocytes from the blood and lymphatics, and their homing to lymph nodes and tumors. L-selectin (CD62L) is an important molecule in these processes. It directs naive lymphocytes to peripheral lymph nodes where they become activated and it traffics naive lymphocytes to inflammatory environments, such as tumors. Individuals with advanced cancer are immune suppressed due to myeloid-derived suppressor cells (MDSC), a population of immature myeloid cells that accumulate to high levels in response to tumor secreted and proinflammatory factors. We now demonstrate that the reduction in T cell levels of L-selectin that is commonly seen in individuals with cancer inversely correlates with MDSC levels. Three lines of evidence demonstrate that MDSC directly down-regulate L-selectin on naive T cells: 1) naive T cells cocultured with tumor-induced MDSC have reduced L-selectin; 2) T cells in tumor free aged mice with elevated levels of MDSC have reduced L-selectin, and 3) peritoneal exudate T cells of tumor-free mice treated with plasminogen activator urokinase to elevate MDSC have reduced levels of L-selectin. MDSC are likely to down-regulate L-selectin through their plasma membrane expression of ADAM17 (a disintegrin and metalloproteinase domain 17), an enzyme that cleaves the ectodomain of L-selectin. Therefore, MDSC down-regulate L-selectin levels on naive T cells, decreasing their ability to home to sites where they would be activated. This is another mechanism by which MDSC inhibit antitumor immunity. PMID- 19553537 TI - Identification of a novel IL-1 cytokine family member in teleost fish. AB - A novel IL-1 family member (nIL-1F) has been discovered in fish, adding a further member to this cytokine family. The unique gene organization of nIL-1F, together with its location in the genome and low homology to known family members, suggests that this molecule is not homologous to known IL-1F. Nevertheless, it contains a predicted C-terminal beta-trefoil structure, an IL-1F signature region within the final exon, a potential IL-1 converting enzyme cut site, and its expression level is clearly increased following infection, or stimulation of macrophages with LPS or IL-1beta. A thrombin cut site is also present and may have functional relevance. The C-terminal recombinant protein antagonized the effects of rainbow trout rIL-1beta on inflammatory gene expression in a trout macrophage cell line, suggesting it is an IL-1beta antagonist. Modeling studies confirmed that nIL-1F has the potential to bind to the trout IL-1RI receptor protein, and may be a novel IL-1 receptor antagonist. PMID- 19553536 TI - Annexin-1 regulates macrophage IL-6 and TNF via glucocorticoid-induced leucine zipper. AB - Annexin-1 (ANXA1) is a mediator of the anti-inflammatory actions of endogenous and exogenous glucocorticoids (GC). The mechanism of ANXA1 effects on cytokine production in macrophages is unknown and is here investigated in vivo and in vitro. In response to LPS administration, ANXA1(-/-) mice exhibited significantly increased serum IL-6 and TNF compared with wild-type (WT) controls. Similarly, LPS-induced IL-6 and TNF were significantly greater in ANXA1(-/-) than in WT peritoneal macrophages in vitro. In addition, deficiency of ANXA1 was associated with impairment of the inhibitory effects of dexamethasone (DEX) on LPS-induced IL-6 and TNF in macrophages. Increased LPS-induced cytokine expression in the absence of ANXA1 was accompanied by significantly increased LPS-induced activation of ERK and JNK MAPK and was abrogated by inhibition of either of these pathways. No differences in GC effects on MAPK or MAPK phosphatase 1 were observed in ANXA1(-/-) cells. In contrast, GC-induced expression of the regulatory protein GILZ was significantly reduced in ANXA1(-/-) cells by silencing of ANXA1 in WT cells and in macrophages of ANXA1(-/-) mice in vivo. GC induced GILZ expression and GC inhibition of NF-kappaB activation were restored by expression of ANXA1 in ANXA1(-/-) cells, and GILZ overexpression in ANXA1(-/-) macrophages reduced ERK MAPK phosphorylation and restored sensitivity of cytokine expression and NF-kappaB activation to GC. These data confirm ANXA1 as a key inhibitor of macrophage cytokine expression and identify GILZ as a previously unrecognized mechanism of the anti-inflammatory effects of ANXA1. PMID- 19553538 TI - Lymphocytes in the peritoneum home to the omentum and are activated by resident dendritic cells. AB - The omentum is of interest in the context of obesity-related metabolic disease where adipose tissue exhibits inflammatory changes; however, the immunology of the omentum is underexplored. The greater omentum is draped from the stomach and consists predominantly of adipose tissue studded with lymphoreticular aggregations (milky spots) that distinguish it from other visceral adipose tissues. Milky spots are thought to contain and conduct leukocytes in transit from the blood to the peritoneal cavity, particularly during peritonitis. We show here that both B and T lymphocytes counterflow from the peritoneal cavity to the omentum in mice. Residence in the omentum was brief with a t(1/2) residence time of 6 h. Omentum access was pertussis toxin-sensitive, dependent on activation of the Rap1 GTPase, and on the integrin LFA-1. B cells and CD44(high) T cells accessed the omentum most efficiently, but homing of resting CD44(low) T cells was also observed. Omental tissue from normal healthy mice was found to contain CD8(-)CD11b(high)MHC class II(high)CD11c(high) dendritic cells that promoted the rapid activation of T cells entering the omentum and cross-presented soluble OVA or OVA acquired from either OVA-expressing Escherichia coli or OVA-pulsed spleen cells. We conclude that the omentum incorporates two key features of immunological sentinel function, actively supported lymphocyte traffic and dendritic cells, that reinforce a conceptual framework for function in stimulating adaptive immunity. These results extend basic understanding of omental and peritoneal cavity immunology and of how proinflammatory events occurring within the peritoneal cavity might affect adipocyte and hepatocyte metabolism. PMID- 19553539 TI - The impact of TCR-binding properties and antigen presentation format on T cell responsiveness. AB - TCR interactions with cognate peptide-MHC (pepMHC) ligands are generally low affinity. This feature, together with the requirement for CD8/CD4 participation, has made it difficult to dissect relationships between TCR-binding parameters and T cell activation. Interpretations are further complicated when comparing different pepMHC, because these can vary greatly in stability. To examine the relationships between TCR-binding properties and T cell responses, in this study we characterized the interactions and activities mediated by a panel of TCRs that differed widely in their binding to the same pepMHC. Monovalent binding of soluble TCR was characterized by surface plasmon resonance, and T cell hybridomas that expressed these TCR, with or without CD8 coexpression, were tested for their binding of monomeric and oligomeric forms of the pepMHC and for subsequent responses (IL-2 release). The binding threshold for eliciting this response in the absence of CD8 (K(D) = 600 nM) exhibited a relatively sharp cutoff between full activity and no activity, consistent with a switchlike response to pepMHC on APCs. However, when the pepMHC was immobilized (plate bound), T cells with the lowest affinity TCRs (e.g., K(D) = 30 microM) responded, even in the absence of CD8, indicating that these TCR are signaling competent. Surprisingly, even cells that expressed high-affinity (K(D) = 16 nM) TCRs along with CD8 were unresponsive to oligomers in solution. The findings suggest that to drive downstream T cell responses, pepMHC must be presented in a form that supports formation of appropriate supramolecular clusters. PMID- 19553540 TI - Insufficient TLR activation contributes to the slow development of CD8+ T cell responses in Trypanosoma cruzi infection. AB - During experimental infection with Trypanosoma cruzi, mice develop a strong CD8(+) T cell response focused mainly on a few immunodominant peptides encoded in trans-sialidase family genes. Despite the potency of this response, the initial emergence and peak of parasite-specific CD8(+) T cells has been noted to be relatively slow. In this study, we further document this delayed onset of T cell responses to T. cruzi as measured by the increase in frequency of parasite specific T cells, the effector function of these cells, T cell proliferation in general, and the recruitment of cells into the draining lymph nodes. This delay does not appear to be the result of general immunosuppressive effects of the infection, a limitation in parasite numbers, or parasite trafficking to lymph nodes or to the specific epitope. Increasing the initial infecting dose or the density of parasite epitopes on APCs can modestly speed the generation of anti-T. cruzi T cell responses. Given these characteristics of the response, we propose that T. cruzi is a stealth invader, largely avoiding recognition by components of the innate immune system until the infection is well established. This conclusion is supported by the ability to accelerate the induction of T cell responses to T. cruzi by administration of ligands for TLR2 and TLR9 at the time of infection. These studies highlight a previously unappreciated mechanism of immune evasion, the surreptitious establishment of infection, by the protozoan T. cruzi. PMID- 19553541 TI - IL-33 enhances lipopolysaccharide-induced inflammatory cytokine production from mouse macrophages by regulating lipopolysaccharide receptor complex. AB - Bacterial LPS triggers monocytes and macrophages to produce several inflammatory cytokines and mediators. However, once exposed to LPS, they become hyporesponsive to a subsequent endotoxin challenge. This phenomenon is defined as LPS desensitization or tolerance. Previous studies have identified some components of the biochemical pathways involved in negative modulation of LPS responses. In particular, it has been shown that the IL-1R-related protein ST2 could be implicated in LPS tolerance. The natural ligand of ST2 was recently identified as IL-33, a new member of the IL-1 family. In this study, we investigated whether IL 33 triggering of ST2 was able to induce LPS desensitization of mouse macrophages. We found that IL-33 actually enhances the LPS response of macrophages and does not induce LPS desensitization. We demonstrate that this IL-33 enhancing effect of LPS response is mediated by the ST2 receptor because it is not found in ST2 knockout mice. The biochemical consequences of IL-33 pretreatment of mouse macrophages were investigated. Our results show that IL-33 increases the expression of the LPS receptor components MD2 (myeloid differentiation protein 2) and TLR-4, the soluble form of CD14 and the MyD88 adaptor molecule. In addition, IL-33 pretreatment of macrophages enhances the cytokine response to TLR-2 but not to TLR-3 ligands. Thus, IL-33 treatment preferentially affects the MyD88 dependent pathway activated by the TLR. PMID- 19553543 TI - Cathepsin S regulates class II MHC processing in human CD4+ HLA-DR+ T cells. AB - Although it has long been known that human CD4(+) T cells can express functional class II MHC molecules, the role of lysosomal proteases in the T cell class II MHC processing and presentation pathway is unknown. Using CD4(+) T cell clones that constitutively express class II MHC, we determined that cathepsin S is necessary for invariant chain proteolysis in T cells. CD4(+)HLA-DR(+) T cells down-regulated cathepsin S expression and activity 18 h after activation, thereby ceasing nascent class II MHC product formation. This blockade resulted in the loss of the invariant chain fragment CLIP from the cell surface, suggesting that like professional APC-CD4(+) HLA-DR(+) cells modulate self-Ag presentation as a consequence of activation. Furthermore, cathepsin S expression and activity, and concordantly cell surface CLIP expression, was reduced in HLA-DR(+) CD4(+) T cells as compared with B cells both in vitro and ex vivo. PMID- 19553544 TI - Significance of N-terminal proteolysis of CCL14a to activity on the chemokine receptors CCR1 and CCR5 and the human cytomegalovirus-encoded chemokine receptor US28. AB - The CC chemokine CCL14a is constitutively expressed in a large variety of tissues and its inactive proform CCL14a(1-74) circulates in high concentrations in plasma. CCL14a(1-74) is converted into CCL14a(9-74) by the proteases urokinase type plasminogen activator and plasmin and is a highly active agonist for the chemokine receptors CCR1 and CCR5. In this study, a new CCL14a analog, CCL14a(12 74), was isolated from blood filtrate. To elucidate the functional role of the N terminus, a panel of N-terminally truncated CCL14a analogs were tested on the receptors CCR1 to CCR5 and on the human cytomegalovirus (HCMV)-encoded chemokine receptor US28. The rank order of binding affinity to these receptors and of the activation of CCR1 and CCR5-mediated intracellular Ca(2+) concentration mobilization is CCL14a(6-74)<(7-74)<(8-74)<<(9-74) = (10-74)>>(11-74)>>(12-74). The almost identical affinities of CCL14a(7-74), CCL14a(9-74), and CCL14a(10-74) for the US28 receptor and the inhibition of US28-mediated HIV infection of 293T cells by all of the N-terminally truncated CCL14a analogs support the promiscuous nature of the viral chemokine receptor US28. In high concentrations, CCL14a(12 74) did reveal antagonistic activity on intracellular Ca(2+) concentration mobilization in CCR1- and CCR5-transfected cells, which suggests that truncation of Tyr(11) might be of significance for an efficient inactivation of CCL14a. A putative inactivation pathway of CCL14a(9-74) to CCL14a(12-74) may involve the dipeptidase CD26/dipeptidyl peptidase IV (DPPIV), which generates CCL14a(11-74), and the metalloprotease aminopeptidase N (CD13), which displays the capacity to generate CCL14a(12-74) from CCL14a(11-74). Our results suggest that the activity of CCL14a might be regulated by stringent proteolytic activation and inactivation steps. PMID- 19553542 TI - SUMO conjugation contributes to immune deviation in nonobese diabetic mice by suppressing c-Maf transactivation of IL-4. AB - It is not clear why the development of protective Th2 cells is poor in type 1 diabetes (T1D). c-Maf transactivates the IL-4 gene promoting Th2 cell development; therefore, abnormalities in c-Maf may contribute to reduced IL-4 production by CD4 cells from nonobese diabetic (NOD) mice. In this study we demonstrate that despite normal expression, c-Maf binds poorly to the IL-4 promoter (IL-4p) in NOD CD4 cells. Immunoblotting demonstrates that c-Maf can be modified at lysine 33 by SUMO-1 (small ubiquitin-like modifier 1). Sumoylation is facilitated by direct interaction with the E2-conjugating enzyme Ubc9 and increases following T cell stimulation. In transfected cells, sumoylation decreases c-Maf transactivation of IL-4p-driven luciferase reporter activity, reduces c-Maf binding to the IL-4p in chromatin immunoprecipitation assays, and enhances c-Maf localization into promyelocytic leukemia nuclear bodies. Sumoylation of c-Maf is increased in NOD CD4 cells as compared with CD4 cells from diabetes-resistant B10.D2 mice, suggesting that increased c-Maf sumoylation contributes to immune deviation in T1D by reducing c-Maf access to and transactivation of the IL-4 gene. PMID- 19553546 TI - Direct antigen presentation and gap junction mediated cross-presentation during apoptosis. AB - MHC class I molecules present peptides from endogenous proteins. Ags can also be presented when derived from extracellular sources in the form of apoptotic bodies. Cross-presentation of such Ags by dendritic cells is required for proper CTL responses. The fate of Ags in cells initiated for apoptosis is unclear as is the mechanism of apoptosis-derived Ag transfer into dendritic cells. Here we show that novel Ags can be generated by caspases and be presented by MHC class I molecules of apoptotic cells. Since gap junctions function until apoptotic cells remodel to form apoptotic bodies, transfer and cross-presentation of apoptotic peptides by neighboring and dendritic cells occurs. We thus define a novel phase in classical Ag presentation and cross-presentation by MHC class I molecules: presentation of Ags created by caspase activities in cells in apoptosis. PMID- 19553545 TI - Class switch recombination efficiency and junction microhomology patterns in Msh2 , Mlh1-, and Exo1-deficient mice depend on the presence of mu switch region tandem repeats. AB - The Msh2 mismatch repair (MMR) protein is critical for class switch recombination (CSR) events that occur in mice that lack the Smu tandem repeat (SmuTR) region (SmuTR(-/-) mice). The pattern of microhomology among switch junction sites in Msh2-deficient mice is also dependent on the presence or absence of SmuTR sequences. It is not known whether these CSR effects reflect an individual function of Msh2 or the function of Msh2 within the MMR machinery. In the absence of the SmuTR sequences, Msh2 deficiency nearly ablates CSR. We now show that Mlh1 or Exo1 deficiencies also eliminate CSR in the absence of the SmuTR. Furthermore, in SmuTR(-/-) mice, deficiencies of Mlh1 or Exo1 result in increased switch junction microhomology as has also been seen with Msh2 deficiency. These results are consistent with a CSR model in which the MMR machinery is important in processing DNA nicks to produce double-stranded breaks, particularly in sequences where nicks are infrequent. We propose that double-stranded break paucity in MMR deficient mice leads to increased use of an alternative joining pathway where microhomologies are important for CSR break ligation. Interestingly, when the SmuTR region is present, deficiency of Msh2 does not lead to the increased microhomology seen with Mlh1 or Exo1 deficiencies, suggesting that Msh2 might have an additional function in CSR. It is also possible that the inability to initiate MMR in the absence of Msh2 results in CSR junctions with less microhomology than joinings that occur when MMR is initiated but then proceeds abnormally due to Mlh1 or Exo1 deficiencies. PMID- 19553547 TI - Propionic acid secreted from propionibacteria induces NKG2D ligand expression on human-activated T lymphocytes and cancer cells. AB - We found that propionic acid secreted from propionibacteria induces expression of the NKG2D ligands MICA/B on activated T lymphocytes and different cancer cells, without affecting MICA/B expression on resting peripheral blood cells. Growth supernatant from propionibacteria or propionate alone could directly stimulate functional MICA/B surface expression and MICA promoter activity by a mechanism dependent on intracellular calcium. Deletion and point mutations further demonstrated that a GC-box motif around -110 from the MICA transcription start site is essential for propionate-mediated MICA promoter activity. Other short chain fatty acids such as lactate, acetate, and butyrate could also induce MICA/B expression. We observed a striking difference in the molecular signaling pathways that regulate MICA/B. A functional glycolytic pathway was essential for MICA/B expression after exposure to propionate and CMV. In contrast, compounds with histone deacetylase-inhibitory activity such as butyrate and FR901228 stimulated MICA/B expression through a pathway that was not affected by inhibition of glycolysis, clearly suggesting that MICA/B is regulated through different molecular mechanisms. We propose that propionate, produced either by bacteria or during cellular metabolism, has significant immunoregulatory function and may be cancer prophylactic. PMID- 19553549 TI - Discriminating foot-and-mouth disease virus-infected and vaccinated animals by use of beta-galactosidase allosteric biosensors. AB - Recombinant beta-galactosidases accommodating one or two different peptides from the foot-and-mouth disease virus (FMDV) nonstructural protein 3B per enzyme monomer showed a drastic enzymatic activity reduction, which mainly affected proteins with double insertions. Recombinant beta-galactosidases were enzymatically reactivated by 3B-specific murine monoclonal and rabbit polyclonal antibodies. Interestingly, these recombinant beta-galactosidases, particularly those including one copy of each of the two 3B sequences, were efficiently reactivated by sera from infected pigs. We found reaction conditions that allowed differentiation between sera of FMDV-infected pigs, cattle, and sheep and those of naive and conventionally vaccinated animals. These FMDV infection-specific biosensors can provide an effective and versatile alternative for the serological distinction of FMDV-infected animals. PMID- 19553550 TI - Innate immune defenses induced by CpG do not promote vaccine-induced protection against foot-and-mouth disease virus in pigs. AB - Emergency vaccination as part of the control strategies against foot-and-mouth disease virus (FMDV) has the potential to limit virus spread and reduce large scale culling. To reduce the time between vaccination and the onset of immunity, immunostimulatory CpG was tested for its capacity to promote early protection against FMDV challenge in pigs. To this end, CpG 2142, an efficient inducer of alpha interferon, was injected intramuscularly. Increased transcription of Mx1, OAS, and IRF-7 was identified as a sensitive measurement of CpG-induced innate immunity, with increased levels detectable to at least 4 days after injection of CpG formulated with Emulsigen. Despite this, CpG combined with an FMD vaccine did not promote protection. Pigs vaccinated 2 days before challenge had disease development, which was at least as acute as that of unvaccinated controls. All pigs vaccinated 7 days before challenge were protected without a noticeable effect of CpG. In summary, our results demonstrate the caution required when translating findings from mouse models to natural hosts of FMDV. PMID- 19553548 TI - Immunogenicity of novel DosR regulon-encoded candidate antigens of Mycobacterium tuberculosis in three high-burden populations in Africa. AB - Increasing knowledge about DosR regulon-encoded proteins has led us to produce novel Mycobacterium tuberculosis antigens for immunogenicity testing in human populations in three countries in Africa to which tuberculosis (TB) is endemic. A total of 131 tuberculin skin test-positive and/or ESAT-6/CFP10-positive, human immunodeficiency virus-negative adult household contacts of active pulmonary TB cases from South Africa (n = 56), The Gambia (n = 26), and Uganda (n = 49) were tested for gamma interferon responses to 7 classical and 51 DosR regulon-encoded M. tuberculosis recombinant protein antigens. ESAT-6/CFP10 fusion protein evoked responses in >75% of study participants in all three countries. Of the DosR regulon-encoded antigens tested, Rv1733c was the most commonly recognized by participants from both South Africa and Uganda and the third most commonly recognized antigen in The Gambia. The four most frequently recognized DosR regulon-encoded antigens in Uganda (Rv1733c, Rv0081, Rv1735c, and Rv1737c) included the three most immunogenic antigens in South Africa. In contrast, Rv3131 induced the highest percentage of responders in Gambian contacts (38%), compared to only 3.4% of Ugandan contacts and no South African contacts. Appreciable percentages of TB contacts with a high likelihood of latent M. tuberculosis infection responded to several novel DosR regulon-encoded M. tuberculosis proteins. In addition to significant similarities in antigen recognition profiles between the three African population groups, there were also disparities, which may stem from genetic differences between both pathogen and host populations. Our findings have implications for the selection of potential TB vaccine candidates and for determining biosignatures of latent M. tuberculosis infection, active TB disease, and protective immunity. PMID- 19553551 TI - Selection and characterization of murine monoclonal antibodies to Staphylococcus aureus iron-regulated surface determinant B with functional activity in vitro and in vivo. AB - In an effort to characterize important epitopes of Staphylococcus aureus iron regulated surface determinant B (IsdB), murine IsdB-specific monoclonal antibodies (MAbs) were isolated and characterized. A panel of 12 MAbs was isolated. All 12 MAbs recognized IsdB in enzyme-linked immunosorbent assays and Western blots; 10 recognized native IsdB expressed by S. aureus. The antigen epitope binding of eight of the MAbs was examined further. Three methods were used to assess binding diversity: MAb binding to IsdB muteins, pairwise binding to recombinant IsdB, and pairwise binding to IsdB-expressing bacteria. Data from these analyses indicated that MAbs could be grouped based on distinct or nonoverlapping epitope recognition. Also, MAb binding to recombinant IsdB required a significant portion of intact antigen, implying conformational epitope recognition. Four MAbs with nonoverlapping epitopes were evaluated for in vitro opsonophagocytic killing (OPK) activity and efficacy in murine challenge models. These were isotype switched from immunoglobulin G1 (IgG1) to IgG2b to potentially enhance activity; however, this isotype switch did not appear to enhance functional activity. MAb 2H2 exhibited OPK activity (> or =50% killing in the in vitro OPK assay) and was protective in two lethal challenge models and a sublethal indwelling catheter model. MAb 13C7 did not exhibit OPK (<50% killing in the in vitro assay) and was protective in one lethal challenge model. Neither MAb 13G11 nor MAb 1G3 exhibited OPK activity in vitro or was active in a lethal challenge model. The data suggest that several nonoverlapping epitopes are recognized by the IsdB-specific MAbs, but not all of these epitopes induce protective antibodies. PMID- 19553552 TI - Age-dependent seroprevalence of Toscana virus in central Italy and correlation with the clinical profile. AB - In order to estimate the antibody prevalence rates for Toscana virus (TOSV) among children and adults, we evaluated the seroprevalence of TOSV in a population (n = 2,737) living in Tuscany during the period of 1999 to 2006. The seroprevalence rate was 19.8% in adults and 5.8% in children, showing an age-dependent increase in TOSV-specific immunity. Meningitis due to TOSV infection was more frequent in adults than in children. PMID- 19553553 TI - Development and technical and clinical validation of a quantitative enzyme-linked immunosorbent assay for the detection of human antibodies to hepatitis B surface antigen in recipients of recombinant hepatitis B virus vaccine. AB - Pending removal from the market of a commercial assay (the AUSAB [Abbott Laboratories] enzyme immunoassay [EIA]) for the determination of antibodies to hepatitis B surface antigen (HBsAg), a new in-house quantitative enzyme-linked immunosorbent assay (ELISA) to measure antibodies against HBsAg (anti-HBs) was developed (anti-HBs in-house). Specific anti-HBs antibodies were sandwiched between the precoated HBsAg ad and ay subtypes purified from plasma from hepatitis B virus (HBV) human carriers and the recombinant HBsAg adw2 subtype tagged with horseradish peroxidase. The assay was calibrated against the 1st International Reference Preparation for anti-hepatitis B immunoglobulin (lot 1977 W1042). Analytical sensitivity and the limit of quantitation were estimated at 0.43 mIU/ml and 2.0 mIU/ml, respectively. Overall reproducibility was 11.86%, and accuracy was estimated to be 94.89%. More than 4,000 samples from seven clinical trials were tested with the anti-HBs in-house assay and compared to results generated with AUSAB EIA and AUSAB radioimmunoassay (RIA). During the technical validation, the anti-HBs in-house assay was compared to the AUSAB RIA as a reference (n = 919). Overall assessment of concordance and Deming's regression analysis were performed. The coefficient of correlation between the AUSAB RIA and anti-HBs in-house assay was 0.9815 with a slope of 0.9187. The overall agreement between anti-HBs in-house and AUSAB RIA was 97.61%, considering the clinical cutoffs at 3.3 mIU/ml and 1.0 mIU/ml for the respective assays. From a clinical perspective, seroprotection rates and anti-HBs geometric mean antibody concentrations for individual studies calculated with either the in-house assay or the reference assays were similar. Conclusions of individual studies were confirmed. The performance characteristics of the in-house assay are acceptable. There is no evidence that use of the new assay would lead to different clinical conclusions from the reference method. PMID- 19553554 TI - Characterization of monoclonal antibodies to Junin virus nucleocapsid protein and application to the diagnosis of hemorrhagic fever caused by South American arenaviruses. AB - Junin virus (JUNV), Machupo virus, Guanarito virus, Sabia virus, and Chapare virus are members of New World arenavirus clade B and are the etiological agents of viral hemorrhagic fevers that occur in South America. In this study, we produced three monoclonal antibodies (MAbs) to the recombinant nucleocapsid protein of JUNV, designated C6-9, C11-12, and E4-2. The specificity of these MAbs was examined by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay, and an epitope-mapping method. Using these MAbs, we developed antigen (Ag) capture ELISA systems. We showed that by using MAb C6-9, JUNV Ag was specifically detected. On the other hand, by using MAb C11-12 or E-4 2, the Ags of all human pathogenic South American arenaviruses were detected. The combined use of these Ag capture ELISA systems in the present study may be useful for the diagnosis of acute-phase viral hemorrhagic fever due to infection by a South American arenavirus. PMID- 19553556 TI - Intracellular infections enhance interleukin-6 and plasminogen activator inhibitor 1 production by cocultivated human adipocytes and THP-1 monocytes. AB - Obesity is associated with a chronic inflammatory state, and adipocyte dysfunction is thought to play a crucial role in this. Infection of adipose tissue may trigger the production of inflammatory cytokines, leading to increased recruitment of macrophages into adipose tissue, which in turn may exacerbate the inflammatory state in obesity. Low-grade inflammation was mimicked in an in vitro coculture model with human adipocytes and THP-1 monocytes. Adipocytes and monocytes were infected with adenovirus, cytomegalovirus (CMV), or influenza A virus. After 48 h, transinfection was evaluated and interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha), adiponectin, and plasminogen activator inhibitor 1 (PAI-1) were measured. IL-6 production was upregulated in cocultures of uninfected adipocytes and THP-1 macrophages in a THP-1 cell number-dependent fashion. IL-6 production by CMV-infected adipocytes was increased relative to that of uninfected adipocytes (P < 0.01). IL-6 production by CMV-infected cocultures was 16- to 37-fold higher than that of uninfected adipocytes (P < 0.001). IL-6 production in influenza A virus-infected cocultures was increased 12 to 20-fold (P < 0.05). Only CMV infection increased levels of PAI-1 in cocultures (fourfold; P < 0.05). Soluble factors produced by THP-1 macrophages rather than by adipocytes were responsible for the increased production of IL-6 in cocultures. Infection of cocultivated human adipocytes and THP-1 monocytes with CMV or influenza A virus led to increased production of IL-6 and PAI-1. Thus, infection of adipose tissue evokes an inflammatory response, leading to adipose tissue dysfunction and subsequent overproduction of IL-6 and PAI-1. This may further compound the atherogenic effects of obesity. PMID- 19553555 TI - Phase I safety and immunogenicity study of a candidate meningococcal disease vaccine based on Neisseria lactamica outer membrane vesicles. AB - Natural immunity to meningococcal disease in young children is associated epidemiologically with carriage of commensal Neisseria species, including Neisseria lactamica. We have previously demonstrated that outer membrane vesicles (OMVs) from N. lactamica provide protection against lethal challenge in a mouse model of meningococcal septicemia. We evaluated the safety and immunogenicity of an N. lactamica OMV vaccine in a phase I placebo-controlled, double-blinded clinical trial. Ninety-seven healthy young adult male volunteers were randomized to receive three doses of either an OMV vaccine or an Alhydrogel control. Subsequently, some subjects who had received the OMV vaccine also received a fourth dose of OMV vaccine, 6 months after the third dose. Injection site reactions were more frequent in the OMV-receiving group, but all reactions were mild or moderate in intensity. The OMV vaccine was immunogenic, eliciting rises in titers of immunoglobulin G (IgG) against the vaccine OMVs, together with a significant booster response, as determined by an enzyme-linked immunosorbent assay. Additionally, the vaccine induced modest cross-reactive immunity to six diverse strains of serogroup B Neisseria meningitidis, including IgG against meningococcal OMVs, serum bactericidal antibodies, and opsonophagocytic activity. The percentages of subjects showing > or =4-fold rises in bactericidal antibody titer obtained were similar to those previously reported for the Norwegian meningococcal OMV vaccine against the same heterologous meningococcal strain panel. In conclusion, this N. lactamica OMV vaccine is safe and induces a weak but broad humoral immune response to N. meningitidis. PMID- 19553557 TI - Lipoprotein lipase and hydrofluoric acid deactivate both bacterial lipoproteins and lipoteichoic acids, but platelet-activating factor-acetylhydrolase degrades only lipoteichoic acids. AB - To identify the Toll-like receptor 2 ligand critically involved in infections with gram-positive bacteria, lipoprotein lipase (LPL) or hydrogen peroxide (H(2)O(2)) is often used to selectively inactivate lipoproteins, and hydrofluoric acid (HF) or platelet-activating factor-acetylhydrolase (PAF-AH) is used to selectively inactivate lipoteichoic acid (LTA). However, the specificities of these chemical reactions are unknown. We investigated the reaction specificities by using two synthetic lipoproteins (Pam(3)CSK(4) and FSL-1) and LTAs from pneumococci and staphylococci. Changes in the structures of the two synthetic proteins and the LTAs were monitored by mass spectrometry, and biological activity changes were evaluated by measuring tumor necrosis factor alpha production by mouse macrophage cells (RAW 264.7) following stimulation. PAF-AH inactivated LTA without reducing the biological activities of Pam(3)CSK(4) and FSL-1. Mass spectroscopy confirmed that PAF-AH monodeacylated pneumococcal LTA but did not alter the structure of either Pam(3)CSK(4) or FSL-1. As expected, HF treatment reduced the biological activity of LTA by more than 80% and degraded LTA. HF treatment not only deacylated Pam(3)CSK(4) and FSL-1 but also reduced the activities of the lipoproteins by more than 60%. Treatment with LPL decreased the biological activities by more than 80%. LPL also removed an acyl chain from the LTA and reduced its activity. Our results indicate that treatment with 1% H(2)O(2) for 6 h at 37 degrees C inactivates Pam(3)CSK(4), FSL-1, and LTA by more than 80%. Although HF, LPL, and H(2)O(2) treatments degrade and inactivate both lipopeptides and LTA, PAF-AH selectively inactivated LTA with no effect on the biological and structural properties of the two lipopeptides. Also, the ability of PAF-AH to reduce the inflammatory activities of cell wall extracts from gram positive bacteria suggests LTA to be essential in inflammatory responses to gram positive bacteria. PMID- 19553558 TI - Autoimmune type 1 diabetes genetic susceptibility encoded by human leukocyte antigen DRB1 and DQB1 genes in Tunisia. AB - Human leukocyte antigen (HLA) class II genes contribute to the genetic susceptibility to type 1 diabetes (T1D), and susceptible alleles and haplotypes were implicated in the pathogenesis of T1D. This study investigated the heterogeneity in HLA class II haplotype distribution among Tunisian patients with T1D. This was a retrospective case control study done in Monastir in central Tunisia. The subjects comprised 88 T1D patients and 112 healthy controls. HLA DRB1 and -DQB1 genotyping was done by PCR-sequence-specific priming. Significant DRB1 and DQB1 allelic differences were seen between T1D patients and controls; these differences comprised DRB1*030101 and DQB1*0302, which were higher in T1D patients than in control subjects, and DRB1*070101, DRB1*110101, DQB1*030101, and DQB1*060101, which were lower in T1D patients than in control subjects. In addition, the frequencies of DRB1*030101-DQB1*0201 and DRB1*040101-DQB1*0302 were higher in T1D patients than in control subjects, and the frequencies of DRB1*070101-DQB1*0201 and DRB1*110101-DQB1*030101 haplotypes were lower in T1D patients than in control subjects. Multiple logistic regression analysis revealed the positive association of DRB1*030101-DQB1*0201 and DRB1*040101-DQB1*0302 and the negative association of only DRB1*070101-DQB1*0201 haplotypes with T1D. Furthermore, a significantly increased prevalence of DRB1*030101-DQB1*0201 homozygotes was seen for T1D subjects than for control subjects. Our results confirm the association of specific HLA-DR and -DQ alleles and haplotypes with T1D in Tunisians. The identification of similar and unique haplotypes in Tunisians compared to other Caucasians highlights the need for evaluating the contribution of HLA class II to the genetic susceptibility to T1D with regard to haplotype usage and also to ethnic origin and racial background. PMID- 19553559 TI - From client to pimp: male violence against female sex workers. AB - The present study explores intimate partner violence (IPV) among female sex workers from the red-light area based in Mumbai, India. Using a grounded theory approach, in-depth interviews were conducted with ten sex workers to explore their experiences of IPV in the context of commercial sex work. Narratives were analyzed and themes constructed. A prevalent theme was the complex development of the male role among sex workers, starting as male clients, becoming intimate partners, and ending as their coercive pimps. In addition, themes were compared to the model of coercion in IPV. The model was generally supported, as sex workers reported exploitation from male partners, followed by coercion and ending with intense IPV victimization. This study draws attention to the drawbacks of criminalization of sex work. PMID- 19553560 TI - ADHD psychosocial treatments: generalization reconsidered. PMID- 19553563 TI - Pseudomonas aeruginosa: can studies in engineered cells tell us why is it such a problem in people with cystic fibrosis? Focus on "Cystic fibrosis transmembrane conductance regulator and caveolin-1 regulate epithelial cell internalization of Pseudomonas aeruginosa". PMID- 19553561 TI - FoxO1 induces apoptosis in skeletal myotubes in a DNA-binding-dependent manner. AB - Recent studies indicate that FoxO transcription factors play an important role in promoting muscle atrophy. To study mechanisms mediating effects of FoxO proteins on muscle wasting, FoxO1-estrogen receptor fusion proteins that are activated by treatment with 4-hydroxytamoxifen (4-OH-T) were stably transfected in C(2)C(12) skeletal myoblasts using the pBABE retroviral system and grown into multinucleated skeletal myotubes. Activation of FoxO1 resulted in significant muscle atrophy, which was accompanied by DNA fragmentation, evidenced by terminal deoxynucleotidyl transferase dUTP-mediated nick end labeling. Cells expressing a DNA-binding-deficient form of FoxO1 also exhibited significant atrophy on FoxO1 activation but no hallmark signs of apoptosis. FoxO1 activation resulted in a significant increase in muscle atrophy F-box (MAFbx)/atrogin-1, muscle-specific RING finger protein 1 (MuRF-1), and Bcl-2-interacting mediator of cell death (Bim) gene expression, with no significant increase in Bcl-2/adenovirus E1B 19 kDa-interacting protein 3 (BNip3) gene expression. Although the ability of FoxO1 to induce MuRF-1 gene expression appeared to be independent of DNA binding, expression of MAFbx/atrogin-1 and Bim was significantly blunted in cells expressing DNA-binding-deficient FoxO1. BNip3 gene expression was significantly elevated in DNA-binding-deficient mutant cells. These findings indicate that FoxO1 promotes skeletal muscle atrophy through induction of proteolytic and apoptotic machinery via DNA-binding-dependent and -independent mechanisms. PMID- 19553562 TI - PGC-1alpha attenuates neointimal formation via inhibition of vascular smooth muscle cell migration in the injured rat carotid artery. AB - Oxidative stress contributes significantly to the migration of vascular smooth muscle cells (VSMCs), the major pathogenic process of vascular diseases, but the mechanism remains unclear. In the present study, we explored the role of peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha), a major regulator of mitochondrial biogenesis and energy balance, in VSMC migration in vitro and in vivo. Overexpression of PGC-1alpha in cultured VSMCs led to a 74.5% reduction of migration activity and mitochondrial ROS generation by the increased expression of antioxidative proteins such as SOD-2 in the mitochondria. The knockdown of PGC-1alpha by specific small interfering (si)RNA markedly augmented VSMC migration activity and greatly reduced mitochondrial antioxidative protein expression. Furthermore, knockdown of SOD-2 expression by siRNA greatly reversed the inhibitory effect of PGC-1alpha overexpression on VSMC migration. In a rat carotid balloon injury model, adenovirus-mediated overexpression of PGC-1alpha greatly reduced neointimal formation (ratio of intima to media: 0.78 +/- 0.09 vs. 1.45 +/- 0.18 in the adenovirus + green fluorescent protein gene- transfected group). Moreover, the expression of SOD-2 was significantly increased in vivo in local vessels after injury in the PGC 1alpha-overexpressing group. These data strongly suggest that PGC-1alpha inhibits VSMC migration and neointimal formation after vascular injury in rats, mainly by upregulating the expression of the mitochondrial antioxidant enzyme SOD-2. PMID- 19553564 TI - Different effects of arsenate and phosphonoformate on P(i) transport adaptation in opossum kidney cells. AB - The main nonhormonal mechanism for controlling inorganic phosphate (P(i)) homeostasis is renal adaptation of the proximal tubular P(i) transport rate to changes in dietary phosphate content. Opossum kidney (OK) cell line is an in vitro renal model that maintains the ability of renal adaptation to the extracellular P(i) concentration. We have studied how two competitive inhibitors of P(i) transport, arsenate [As(V)] and phosphonoformate (PFA), affect adaptation to low and high P(i) concentrations. OK cells show very high affinity for As(V) (inhibitory constant, K(i) 0.12 mM) when compared with the rat kidney. As(V) very efficiently reversed the adaptation of OK cells to low P(i) (0.1 mM), whereas PFA induced adaptation similar to 0.1 mM P(i). Adaptation with 2 mM P(i) or As(V) was characterized by decreases in the maximal velocity (V(max)) of P(i) transport and an abundance of the NaPi-IIa P(i) transporter in the plasma membrane, shown by the protein biotinylation. Conversely, PFA and 0.1 mM P(i) increased the V(max) and transporter abundance. Changes in the V(max) were limited to a 50% variation, which was not paralleled by changes in the concentration of P(i) or of the inhibitor. OK cells are very sensitive to As(V), but the effects are reversible and noncytotoxic. These effects can be interpreted as As(V) being transported into the cell, thereby mimicking a high P(i) concentration. PFA blocks the uptake of P(i) but is not transported, and it therefore simulates a low P(i) concentration inside the cell. To conclude, a mathematical definition of the adaptation process is reported, thereby explaining the limited changes in P(i) transport V(max). PMID- 19553565 TI - Effect of resveratrol and zinc on intracellular zinc status in normal human prostate epithelial cells. AB - To evaluate the influence of resveratrol on cellular zinc status, normal human prostate epithelial (NHPrE) cells were treated with resveratrol (0, 0.5, 1, 2.5, 5, and 10 microM) and zinc [0, 4, 16, and 32 microM, representing zinc-deficient (ZD), zinc-normal (ZN), zinc-adequate (ZA), and zinc-supplemented (ZS) conditions, respectively]. A progressive reduction in cell growth was observed in cells treated with increasing amounts of resveratrol (2.5-10 microM). Resveratrol at 5 and 10 microM resulted in a dramatic increase in cellular total zinc concentration, especially in ZS cells. Flow cytometry indicated that 10 microM resveratrol induced arrest of the cell cycle at the G(2)/M phase in association with the observed cell growth inhibition. Data from an in vitro experiment using zinquin as an indicator of intracellular free Zn(II) status demonstrated complex interactions between resveratrol and Zn(II). Fluorescence spectrofluorometry and fluorescence microscopic analyses revealed that intracellular free labile zinc was progressively elevated from nearly twofold in ZS cells with no resveratrol to multifold in ZA and ZS cells with 10 microM resveratrol compared with the corresponding ZN cells. Furthermore, increases in cellular zinc status were associated with elevated levels of reactive oxygen species and senescence, as evidenced by morphological and histochemical changes in cells treated with 2.5 or 10 microM resveratrol, especially in ZA and ZS cells. Taken together, the interaction between resveratrol and zinc in NHPrE cells increases total cellular zinc and intracellular free labile zinc status and, subsequently, reactive oxygen species production and senescence. PMID- 19553566 TI - Inhibition of CDKS by roscovitine suppressed LPS-induced *NO production through inhibiting NFkappaB activation and BH4 biosynthesis in macrophages. AB - In inflammatory diseases, tissue damage is critically associated with nitric oxide ((*)NO) and cytokines, which are overproduced in response to cellular release of endotoxins. Here we investigated the inhibitory effect of roscovitine, a selective inhibitor of cyclin-dependent kinases (CDKs) on (*)NO production in mouse macrophages. In RAW264.7 cells, we found that roscovitine abolished the production of (*)NO induced by lipopolysaccharide (LPS). Moreover, roscovitine significantly inhibited LPS-induced inducible nitric oxide synthase (iNOS) mRNA and protein expression. Our data also showed that roscovitine attenuated LPS induced phosphorylation of IkappaB kinase beta (IKKbeta), IkappaB, and p65 but enhanced the phosphorylation of ERK, p38, and c-Jun NH(2)-terminal kinase (JNK). In addition, roscovitine dose dependently inhibited LPS-induced expression of cyclooxygenase-2 (COX)-2, IL-1beta, and IL-6 but not tumor necrosis factor (TNF) alpha. Tetrahydrobiopterin (BH(4)), an essential cofactor for iNOS, is easily oxidized to 7,8-dihydrobiopterin (BH(2)). Roscovitine significantly inhibited LPS induced BH(4) biosynthesis and decreased BH(4)-to-BH(2) ratio. Furthermore, roscovitine greatly reduced the upregulation of GTP cyclohydrolase-1 (GCH-1), the rate-limiting enzyme for BH(4) biosynthesis. Using other CDK inhibitors, we found that CDK1, CDK5, and CDK7, but not CDK2, significantly inhibited LPS-induced (*)NO production in macrophages. Similarly, in isolated peritoneal macrophages, roscovitine strongly inhibited (*)NO production, iNOS, and COX-2 upregulation, activation of NFkappaB, and induction of GCH-1 by LPS. Together, our data indicate that roscovitine abolishes LPS-induced (*)NO production in macrophages by suppressing nuclear factor-kappaB activation and BH(4) biosynthesis, which might be mediated by CDK1, CDK5, and CDK7. Our results also suggest that roscovitine may inhibit inflammation and that CDKs may play important roles in the mechanisms by which roscovitine attenuates inflammation. PMID- 19553567 TI - Functional analysis of human RhCG: comparison with E. coli ammonium transporter reveals similarities in the pore and differences in the vestibule. AB - Rh glycoproteins are members of the ammonium transporter (Amt)/methylamine permease (Mep)/Rh family facilitating movement of NH(3) across plasma membranes. Homology models constructed on the basis of the experimental structures of Escherichia coli AmtB and Nitrosomonas europaea Rh50 indicated a channel structure for human RhA (RhAG), RhB (RhBG), and RhC (RhCG) glycoproteins in which external and internal vestibules are linked by a pore containing two strictly conserved histidines. The pore entry is constricted by two highly conserved phenylalanines, "twin-Phe." In this study, RhCG function was investigated by stopped-flow spectrofluorometry measuring kinetic pH variations in HEK293E cells in the presence of an ammonium gradient. The apparent unitary NH(3) permeability of RhCG was determined and was found to be close to that of AmtB. With a site directed mutagenesis approach, critical residues involved in Rh NH(3) channel activity were highlighted. In the external vestibule, the importance of both the charge and the conformation of the conserved aspartic acid was shown. In contrast to AmtB, individual mutations of each phenylalanine of the twin-Phe impaired the function while the removal of both resulted in recovery of the transport activity. The impact of the mutations suggests that, although having a common function and a similar channel structure, bacterial AmtB and human Rh vary in several aspects of the NH(3) transport mechanisms. PMID- 19553568 TI - Impaired ATP turnover and ADP supply depress cardiac mitochondrial respiration and elevate superoxide in nonfailing spontaneously hypertensive rat hearts. AB - Although most attention has been focused on mitochondrial ATP production and transfer in failing hearts, less has been focused on the nonfailing hypertensive heart. Here, energetic complications are less obvious, yet they may provide insight into disease ontogeny. We studied hearts from 12-mo-old spontaneously hypertensive rats (SHR) relative to normotensive Wistar-Kyoto (WKY) rats. The ex vivo working-heart model of SHR showed reduced compliance and impaired responses to increasing preloads. High-resolution respirometry showed higher state 3 (with excess ADP) respiration in SHR left ventricle fibers with complex I substrates and maximal uncoupled respiration with complex I + complex II substrates. Respiration with ATP was depressed 15% in SHR fibers relative to WKY fibers, suggesting impaired ATP hydrolysis. This finding was consistent with a 50% depression of actomyosin ATPase activities. Superoxide production from SHR fibers was similar to that from WKY fibers respiring with ADP; however, it was increased by 15% with ATP. In addition, the apparent K(m) for ADP was 54% higher for SHR fibers, and assays conducted after ex vivo work showed a 28% depression of complex I in SHR, but not WKY, fibers. Transmission electron microscopy showed similar mitochondrial volumes but a decrease in the number of cristae in SHR mitochondria. Tissue lipid peroxidation was also 15% greater in SHR left ventricle. Overall, these data suggest that although cardiac mitochondria from nonfailing SHR hearts function marginally better than those from WKY hearts, they show dysfunction after intense work. Impaired ATP turnover in hard-working SHR hearts may starve cardiac mitochondria of ADP and elevate superoxide. PMID- 19553569 TI - Unexpectedly high proportion of ancestral Manu genotype Mycobacterium tuberculosis strains cultured from tuberculosis patients in Egypt. AB - Tuberculosis is one of the important public health problems in Egypt. However, limited information on the Mycobacterium tuberculosis genotypes circulating in Egypt is available. A total of 151 M. tuberculosis strains were characterized by spoligotyping. The results revealed that 74.8% of M. tuberculosis isolates grouped into 13 different clusters, while 25.2% had unique spoligotype patterns. Comparison with an international spoligotyping database (the SITVIT2 database) showed that types SIT53 (T1 variant) and SIT54 (Manu2 variant) were the most common types between cluster groups. In addition, new shared types SIT2977, SIT2978, and SIT2979 were observed. The results identified for the first time an unusually high proportion of ancestral Manu strains of M. tuberculosis from patients in Egypt. The percentage of the Manu clade in this study (27.15%) was significantly higher than its overall representation of 0.4% in the SITVIT2 database. We show that in Egypt tuberculosis is caused by a predominant M. tuberculosis genotype belonging to the ancestral Manu lineage which could be a missing link in the split between ancestral and modern tubercle bacilli during the evolution of M. tuberculosis. PMID- 19553570 TI - Detection and genotyping of human papillomavirus in self-obtained cervicovaginal samples by using the FTA cartridge: new possibilities for cervical cancer screening. AB - This study assesses human papillomavirus (HPV) detection and genotyping in self sampled genital smears applied to an indicating FTA elute cartridge (FTA cartridge). The study group consisted of 96 women, divided into two sample sets. All samples were analyzed by the HPV SPF(10)-Line Blot 25. Set 1 consisted of 45 women attending the gynecologist; all obtained a self-sampled cervicovaginal smear, which was applied to an FTA cartridge. HPV results were compared to a cervical smear (liquid based) taken by a trained physician. Set 2 consisted of 51 women who obtained a self-sampled cervicovaginal smear at home, which was applied to an FTA cartridge and to a liquid-based medium. DNA was obtained from the FTA cartridges by simple elution as well as extraction. Of all self-obtained samples of set 1, 62.2% tested HPV positive. The overall agreement between self- and physician-obtained samples was 93.3%, in favor of the self-obtained samples. In sample set 2, 25.5% tested HPV positive. The overall agreement for high-risk HPV presence between the FTA cartridge and liquid-based medium and between DNA elution and extraction was 100%. This study shows that HPV detection and genotyping in self-obtained cervicovaginal samples applied to an FTA cartridge is highly reliable. It shows a high level of overall agreement with HPV detection and genotyping in physician-obtained cervical smears and liquid-based self samples. DNA can be obtained by simple elution and is therefore easy, cheap, and fast. Furthermore, the FTA cartridge is a convenient medium for collection and safe transport at ambient temperatures. Therefore, this method may contribute to a new way of cervical cancer screening. PMID- 19553572 TI - Evaluation of MeningoFinder, a novel multiplex ligation-dependent probe amplification assay for simultaneous detection of six virus species causing central nervous system infections. AB - A multiplex ligation-dependent probe amplification assay for simultaneous detection of six virus species was developed and tested on clinical cerebrospinal fluid (CSF) samples. The assay, termed MeningoFinder, showed an accordance of 97%, concordance of 96%, interlaboratory sensitivity of 90%, and interlaboratory specificity of 94% compared to PCRs. PMID- 19553571 TI - Proteomic identification of OprL as a seromarker for initial diagnosis of Pseudomonas aeruginosa infection of patients with cystic fibrosis. AB - Identification of new immunogenic antigens that diagnose initial Pseudomonas aeruginosa infections in patients with cystic fibrosis (CF) alone or as an adjunct to microbiology is needed. In the present study, a proteomic analysis was performed to obtain a global assessment of the host immune response during the initial P. aeruginosa infection of patients with CF. Matrix-assisted laser desorption ionization-time of flight mass spectrometry was used to identify outer membrane protein L (OprL), a non-type III secretion system (TTSS) protein, as an early immunogenic protein during the initial P. aeruginosa infection of patients with CF. Longitudinal Western blot analysis of sera from 12 of 14 patients with CF detected antibodies to OprL during the initial P. aeruginosa infection. In addition, also detected were antibodies to ExoS, ExoU, or ExoS and ExoU, the latter indicating sequential P. aeruginosa infections during initial infections. Detection of serum reactivity to OprL, along with proteins of the TTSS, and in conjunction with microbiology may diagnose initial P. aeruginosa infections in patients with CF. PMID- 19553573 TI - Aeromonas salmonicida subsp. salmonicida type strain does not possess a type III secretion system. PMID- 19553574 TI - Distribution of lag-1 alleles and sequence-based types among Legionella pneumophila serogroup 1 clinical and environmental isolates in the United States. AB - Approximately 84% of legionellosis cases are due to Legionella pneumophila serogroup 1. Moreover, a majority of L. pneumophila serogroup 1 clinical isolates react positively with monoclonal antibody 2 (MAb2) of the international standard panel. Over 94% of the legionellosis outbreaks investigated by the Centers for Disease Control and Prevention are due to this subset of L. pneumophila serogroup 1. To date, there is no complete explanation for the enhanced ability of these strains to cause disease. To better characterize these organisms, we subtyped 100 clinical L. pneumophila serogroup 1 isolates and 50 environmental L. pneumophila serogroup 1 isolates from the United States by (i) reactivity with MAb2, (ii) presence of a lag-1 gene required for the MAb2 epitope, and (iii) sequence-based typing analysis. Our results showed that the MAb2 epitope and lag-1 gene are overrepresented in clinical L. pneumophila serogroup 1 isolates. MAb2 recognized 75% of clinical isolates but only 6% of environmental isolates. Similarly, 75% of clinical isolates but only 8% of environmental isolates harbored lag-1. We identified three distinct lag-1 alleles, referred to as Philadelphia, Arizona, and Lens alleles, among 79 isolates carrying this gene. The Arizona allele is described for the first time in this study. We identified 59 different sequence types (STs), and 34 STs (58%) were unique to the United States. Our results support the hypothesis that a select group of STs may have an enhanced ability to cause legionellosis. Combining sequence typing and lag-1 analysis shows that STs tend to associate with a single lag-1 allele type, suggesting a hierarchy of virulence genotypes. Further analysis of ST and lag-1 profiles may identify genotypes of L. pneumophila serogroup 1 that warrant immediate intervention. PMID- 19553575 TI - Detection and genotyping of human rotavirus VP4 and VP7 genes by reverse transcriptase PCR and reverse hybridization. AB - Rotavirus infections can be diagnosed in stool samples by serological and molecular methods. We developed a novel reverse transcriptase PCR (RT-PCR) method for the amplification of rotavirus RNA and a reverse hybridization assay on a strip to detect amplimers and identify the specific G and P genotypes present in human stool specimens. An additional aim was to permit specific identification of the rotavirus G1P[8] strain, used in the Rotarix vaccine. Novel broad-spectrum PCR primers were developed for both VP4 and VP7, permitting the amplification of a wide range of rotavirus genotypes. Primer sets comprise mixtures of defined primer sequences. For the identification of G and P genotypes, two reverse hybridization strip assays were developed. Both the VP4 and the VP7 strip contain universal probes for the detection of VP4 and VP7 sequences, irrespective of the G or P genotype. The VP4 strip contains type-specific probes for P[4], P[6], P[8], P[9], and P[10]. The VP7 strip contains type-specific probes for G1, G2, G3, G4, G5, G6, G8, and G9. In addition, probes to distinguish between wild-type G1 and G1 vaccine strain sequences were present. Testing by analysis of multiple reference strains confirmed that both RT-PCR methods allowed the detection of a broad spectrum of genotypes. RT-PCR for VP7 was more sensitive than RT-PCR for VP4, but all samples identified as positive for rotavirus antigen by an enzyme linked immunosorbent assay (ELISA) were also positive for both VP4 and VP7. The high specificity of the reverse hybridization method was confirmed by sequence analysis as well as by type-specific PCR, and the vaccine strain could also be specifically identified. The reverse hybridization method permits accurate identification of mixed infections with different genotypes. Rotavirus genotypes for which no type-specific probes were present on the strip were adequately identified by the universal detection probes. The assay was formally validated by analyses of specificity, sensitivity, precision, accuracy, and robustness. In a panel of 149 ELISA-positive stool samples, comparison with conventional type specific RT-PCR methods revealed the superiority of the novel method, mainly in cases of mixed rotavirus infections. This novel method permits highly accurate detection and identification of human rotavirus infections in stool samples. This validated assay could be useful for large-scale epidemiological and clinical trials. PMID- 19553576 TI - Emerging genotypes of human respiratory syncytial virus subgroup A among patients in Japan. AB - Human respiratory syncytial virus (HRSV) is a common etiological agent of acute lower respiratory tract disease in infants. We report the molecular epidemiology of HRSV in Niigata, Japan, over six successive seasons (from 2001 to 2007) and the emerging genotypes of HRSV subgroup A (HRSV-A) strains. A total of 488 HRSV samples were obtained from 1,103 screened cases in a pediatric clinic in Niigata. According to the phylogenetic analysis, among the PCR-positive samples, 338 HRSV A strains clustered into the previously reported genotypes GA5 and GA7 and two novel genotypes, NA1 and NA2, which were genetically close to GA2 strains. One hundred fifty HRSV-B strains clustered into three genotypes, namely, GB3, SAB3, and BA, which has a 60-nucleotide insertion in the second hypervariable region of the G protein. The NA1 strains emerged first, in the 2004-2005 season, and subsequently, the NA2 strain emerged in the 2005-2006 season. Both strains caused large epidemics in the 2005-2006 and 2006-2007 seasons. The average age of children who were infected with NA2 strains was significantly higher than that of those infected with GA5 and the frequency of reinfection by NA2 was the highest among all genotypes, suggesting that this genotype possessed new antigenicity for evading past host immunity. This is the first paper to show a possible correlation between an emerging genotype, NA2, and large outbreaks of HRSV in Japan. Continuing studies to follow up the genetic changes and to clarify the mechanism of reinfection in HRSV are important steps to understand HRSV infections. PMID- 19553578 TI - Glans swabs are not appropriate specimens for diagnosis of Chlamydia trachomatis infection in asymptomatic men. PMID- 19553577 TI - Emergence of a clonal lineage of Brucella abortus biovar 3 in clinical cases in Spain. PMID- 19553579 TI - Leishmania OligoC-TesT as a simple, rapid, and standardized tool for molecular diagnosis of cutaneous leishmaniasis in Peru. AB - Molecular methods such as PCR have become attractive tools for diagnosis of cutaneous leishmaniasis (CL), both for their high sensitivity and for their specificity. However, their practical use in routine diagnosis is limited due to the infrastructural requirements and the lack of any standardization. Recently, a simplified and standardized PCR format for molecular detection of Leishmania was developed. The Leishmania OligoC-TesT is based on simple and rapid detection using a dipstick with PCR-amplified Leishmania DNA. In this study, we estimated the diagnostic accuracy of the Leishmania OligoC-TesT for 61 specimens from 44 CL suspected patients presenting at the leishmaniasis clinic of the Instituto de Medicina Tropical Alexander von Humboldt, Peru. On the basis of parasitological detection and the leishmanin skin test (LST), patients were classified as (i) confirmed CL cases, (ii) LST-positive cases, and (iii) LST-negative cases. The sensitivities of the Leishmania OligoC-TesT was 74% (95% confidence interval (CI), 60.5% to 84.1%) for lesion aspirates and 92% (95% CI, 81.2% to 96.9%) for scrapings. A significantly higher sensitivity was observed with a conventional PCR targeting the kinetoplast DNA on the aspirates (94%) (P = 0.001), while there was no significant difference in sensitivity for the lesion scrapings (88%) (P = 0.317). In addition, the Leishmania OligoC-TesT was evaluated for 13 CL-suspected patients in two different peripheral health centers in the central jungle of Peru. Our findings clearly indicate the high accuracy of the Leishmania OligoC TesT for lesion scrapings for simple and rapid molecular diagnosis of CL in Peru. PMID- 19553580 TI - Evaluation of GenoType mycobacteria direct assay in comparison with Gen-Probe Mycobacterium tuberculosis amplified direct test and GenoType MTBDRplus for direct detection of Mycobacterium tuberculosis complex in clinical samples. AB - Three molecular assays were evaluated for the direct detection of Mycobacterium tuberculosis complex bacteria in 125 respiratory and 22 nonrespiratory samples. The overall sensitivities obtained were as follows: GenoType MTBDRplus, 97.9%; GenoType Mycobacteria Direct, 93.7%; Gen-Probe Mycobacterium tuberculosis Amplified Direct Test, 89.6%. The specificity of the assays used was 100%. PMID- 19553581 TI - Plasmid-borne virulence-associated genes have a conserved organization in virulent strains of avian pathogenic Escherichia coli. AB - Avian pathogenic Escherichia coli (APEC) is an important respiratory pathogen of poultry. Various virulence factors are responsible for determining the pathogenicity of these strains, and it is commonly believed they are encoded on large plasmids the strains carry. This study examined a series of strains, the pathogenicity of which had previously been determined by aerosol exposure, for possession of large plasmids and found all isolates carried at least one large plasmid, regardless of the level of virulence. Virulence-associated genes carried on these plasmids were also examined, and it was shown that highly virulent strains carried at least four virulence-associated genes on their largest plasmid. Two of the virulence-associated genes were shown to be chromosomally located in a strain of intermediate virulence, while no virulence-associated genes were carried by the low-virulence strain. The organization of the virulence associated genes was shown to be highly conserved among APEC isolates of high virulence, supporting the concept of a conserved portion of the putative virulence region that contributes to the pathogenicity of APEC strains. PMID- 19553583 TI - Quantitative detection of the M204V hepatitis B virus minor variants by amplification refractory mutation system real-time PCR combined with molecular beacon technology. AB - Mutations in the highly conserved tyrosine-methionine-aspartate-aspartate (YMDD) motif are frequently associated with resistance to antivirals and represent a major concern in the treatment of hepatitis B virus (HBV) infection. Conventional methods fail to detect minority populations of drug-resistant viral quasispecies if they represent less than 25% of the total sample virus population. The amplification refractory mutation system real-time PCR (ARMS RT-PCR) was combined with molecular beacon technology using the LightCycler system. The samples from HBV patients selected for assay evaluation included (i) 57 samples from treatment naive patients for biological discriminatory ability (cutoff) estimation, (ii) 12 samples from patients with treatment failure that were M204V positive by sequencing, and (iii) 13 samples from patients with treatment failure that were negative for mutation at codon 204 by sequencing. The discriminatory ability of the assay was 0.25% when tested with laboratory-synthesized DNA target sequences. The median mutant-to-wild-type ratio for samples from naive patients tested positive for the wild type and for mutant variants was 0.01% (5th and 95th percentiles = 0.0001 and 0.04%, respectively). A value of 0.04% was selected as the biological cutoff of the assay of clinical samples. In all samples M204V positive by sequencing (12/12), the mutant variant was detected as the predominant population (range, 82.76 to 99.43%). Interestingly, in 5 (38%) of 13 samples negative by sequencing, the M204V variant was detected at a ratio above the biological cutoff (0.05 to 28%). The assay represents an efficient technique for the early detection and quantification of M204V variants before mutant strains emerge to dominate the population. PMID- 19553582 TI - Duplex PCR assay simultaneously detecting and differentiating Bartonella quintana, B. henselae, and Coxiella burnetii in surgical heart valve specimens. AB - A duplex PCR (dPCR) assay was developed to simultaneously detect and differentiate Bartonella quintana, Bartonella henselae, and Coxiella burnetii from surgical heart valve tissue specimens with an analytic sensitivity of 10 copies/reaction. Among 17 specimens collected from patients with a clinical diagnosis of culture-negative endocarditis, 2, 4, and 2 were positive for B. quintana, B. henselae, and C. burnetii, respectively, by the dPCR assay, which matched the results obtained by universal bacterial 16S rRNA gene amplification and sequencing. PMID- 19553584 TI - Use of a new single multiplex PCR-based assay for direct simultaneous characterization of six Neisseria meningitidis serogroups. AB - We developed a new Neisseria meningitidis multiplex PCR to determine six serogroups, including X-specific primers, and to allow direct W135/Y discrimination. This assay offers a simple and low-cost method for serogrouping N. meningitidis from cerebrospinal fluid that could be useful in Africa. PMID- 19553585 TI - Emergence of European avian influenza virus-like H1N1 swine influenza A viruses in China. AB - During swine influenza surveillance from 2007 to 2008, 10 H1N1 viruses were isolated and analyzed for their antigenic and phylogenetic properties. Our study revealed the emergence of avian-origin European H1N1 swine influenza virus in China, which highlights the necessity of swine influenza surveillance for potential pandemic preparedness. PMID- 19553587 TI - Lack of heteroresistance among Staphylococcus aureus isolates with vancomycin MICs of 2 micrograms per milliliter by automated testing. PMID- 19553586 TI - Epidemiology and phylogenetic analysis of crimean-congo hemorrhagic fever viruses in xinjiang, china. AB - In 2004 and 2005, an epidemiological survey of Crimean-Congo hemorrhagic fever virus (CCHFV) was conducted in Xinjiang, China. A total of 5,629 serum samples of human and livestock were collected and tested for the CCHFV antibody, and 17,319 ticks were collected for viral identification. Reverse passive hemagglutination inhibition assays showed that the average prevalence of CCHFV antibody was 1.7% for the humans and 12.7% for the livestock. A relatively high antibody prevalence, ranging from 19.1% to 23.4%, was found in the livestock of the northwest, southwest, and northeast parts of the Tarim Basin. When the ticks were pooled to inoculate suckling mice, followed by reverse transcription-PCR (RT-PCR) to detect CCHFV RNA, the average RT-PCR-positive rates for Hyalomma asiaticum kozlovi and H. asiaticum asiaticum were 12.9% and 2.6%, respectively. A significant correlation was found between the antibody prevalence in the livestock and the CCHFV prevalence in H. asiaticum of the same geographic region. No CCHFV RNA was detected in Dermacentor nivenus, Rhipicephalus turanius, or Rhipicephalus sanguineus. A total of 27 partial S segments of CCHFVs were sequenced and used for phylogeny analysis. All but one Chinese isolate grouped into the Asia 1 clade, which contains the strains from Xinjiang and Uzbekistan, while the other strain, Fub90009, grouped with strains from the Middle East. PMID- 19553588 TI - Comparison of typing results obtained for methicillin-resistant Staphylococcus aureus isolates with the DiversiLab system and pulsed-field gel electrophoresis. AB - We compared the results of typing methicillin-resistant Staphylococcus aureus (MRSA) isolates using the DiversiLab system (DL) to the results obtained using pulsed-field gel electrophoresis (PFGE). One hundred five MRSA isolates of PFGE types USA100 to USA1100 and the Brazilian clone, from the Centers for Disease Control and Prevention (CDC) and Project ICARE strain collections, were typed using DL. In addition, four unique sets of MRSA isolates from purported MRSA outbreaks that had been previously typed by DL, each consisting of six isolates (where five isolates were classified as indistinguishable by DL and one was an unrelated DL type) were typed by PFGE. DL separated the 105 MRSA isolates of known USA types into 11 clusters and six unique banding patterns. DL grouped most of the USA100, USA200, and USA1100 isolates into unique clusters. Multilocus sequence type 8 isolates (i.e., USA300 and USA500) often clustered together at >95% similarity in DL dendrograms. Nevertheless, USA300 and USA500 DL patterns could be distinguished using the pattern overlay function of the DL software. Among the hospital outbreak clusters, PFGE and DL identified the same "unrelated" organism in three of four sets. However, PFGE showed more pattern diversity than did DL, suggesting that two of the sets were less likely to represent true outbreaks. In summary, DL is useful for screening MRSA isolates to rule out potential outbreaks of MRSA in hospitals, but PFGE provides better discrimination of potential outbreak strains and is more useful for confirming strain relatedness and specific USA types. PMID- 19553589 TI - Detection of novel (swine origin) H1N1 influenza A virus by quantitative real time reverse transcription-PCR. PMID- 19553590 TI - Will submandibular TCD prevent stroke in children with sickle cell anemia? PMID- 19553591 TI - HIV-associated brain dysfunction in the era of HAART: reasons for hope, but continued concern. PMID- 19553592 TI - Migraine and risk of cardiovascular disease in women: learning about relative and absolute risk. PMID- 19553593 TI - Submandibular TCD approach detects post-bulb ICA stenosis in children with sickle cell anemia. AB - BACKGROUND: Transcranial Doppler (TCD) ultrasound is a procedure commonly used to screen individuals with the major hemoglobin S diseases, Hb SS and Hb S-beta(0), for significant stenoses in the circle of Willis. Flow velocities above 200 cm/s have been shown to identify patients at elevated risk for cerebral infarction. Among TCD's limitations is the inability to insonate the distal extracranial, petrous, and cavernous internal carotid artery (ICA) through the standard transtemporal approach. METHODS: We extended the submandibular approach to include infra-siphon portions of the ICA. RESULTS: Using the extended submandibular approach to evaluate these portions of the ICA, we identified stenotic lesions in 4 patients with Hb SS disease out of a population of 131 children with Hb SS. Three of the 4 patients had no history of overt stroke or stroke-like symptoms. Neuroimaging confirmed the stenotic lesions, and also revealed watershed infarction as well as discrete areas of silent infarction. All 4 children had neuropsychological impairment. CONCLUSIONS: The submandibular approach, when added to a standard transcranial Doppler examination, may increase the sensitivity of this technique to identify important potential sources of cerebral infarction. PMID- 19553594 TI - Migraine frequency and risk of cardiovascular disease in women. AB - BACKGROUND: Migraine has been associated with risk of cardiovascular disease (CVD). Data on the association between migraine frequency and CVD are sparse. METHODS: Prospective cohort study of 27,798 US women aged >or=45 years, who were free of CVD, and for whom we had information on lipids and migraine frequency. We categorized migraine frequency as < monthly, monthly, and >or= weekly. Incident CVD was confirmed after medical record review. RESULTS: Of the 3,568 women with active migraine at baseline, 75.3% reported a migraine frequency of < monthly, 19.7% monthly, and 5.0% >or= weekly. During 11.9 years of follow-up, 706 CVD events occurred. Compared with women without migraine, the multivariable-adjusted hazard ratios (HRs) (95% confidence intervals) among active migraineurs for CVD were 1.55 (1.22-1.97), 0.65 (0.31-1.38), and 1.93 (0.86-4.33) for an attack frequency of < monthly, monthly, and >or= weekly, respectively. The association between migraine frequency and CVD was only apparent among migraineurs with aura. Among those, the multivariable-adjusted HRs for women with a migraine frequency < monthly ranged from 1.81 (1.30-2.50) for coronary revascularizations to 2.43 (1.58-3.74) for myocardial infarction. For women with active migraine with aura and migraine frequencies of >or= weekly, we only found significant increased risk of ischemic stroke (HR = 4.25 [1.36-13.29]). CONCLUSIONS: In our data, the association between migraine and cardiovascular disease varies by migraine frequency. Significant associations were only found among women with migraine with aura. Ischemic stroke was the only outcome associated with a high-attack frequency while a low-attack frequency was associated with any vascular event. Low number of outcome events should caution the interpretation. PMID- 19553595 TI - Regulation of gene expression by estrogen and testosterone in the proximal mouse reproductive tract. AB - The role of estrogen and testosterone in the regulation of gene expression in the proximal reproductive tract is not completely understood. To address this question, mice were treated with testosterone or estradiol, and RNA from the efferent ducts and caput epididymides was processed and hybridized to Affymetrix M430 2.0 microarrays. Analysis of array output identified probe sets in each tissue with altered levels in hormone-treated versus control animals. Hormone treatment efficacy was confirmed by determination of serum hormone levels before and after treatment and by observed changes in transcript levels of previously reported hormone-responsive genes. Tissue-specific hormone sensitivity was observed with 2867 and 3197 probe sets changing significantly in the efferent ducts after estrogen and testosterone treatment, respectively. In the caput epididymidis, 117 and 268 probe sets changed after estrogen and testosterone treatment, respectively, demonstrating a greater response to hormone in the efferent ducts than in the caput epididymidis. Transcripts sharing similar profiles in the intact and hormone-treated animals compared with castrated controls were also identified. Ontology analysis of probe sets revealed that a significant number of hormone-regulated transcripts encode proteins associated with lipid metabolism, transcription, and steroid metabolism in both tissues. Real-time RT-PCR was used to confirm array data and to investigate other potential hormone-responsive regulators of proximal reproductive tract function. The results of this work reveal previously unknown responses to estrogen in the caput epididymidis and to testosterone in the efferent ducts, as well as tissue specific hormone sensitivity in the proximal reproductive tract. PMID- 19553596 TI - Fibroblast growth factors and epidermal growth factor cooperate with oocyte derived members of the TGFbeta superfamily to regulate Spry2 mRNA levels in mouse cumulus cells. AB - Mouse oocytes produce members of the transforming growth factor beta (TGFbeta) superfamily, including bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9), as well as fibroblast growth factors (FGFs). These growth factors cooperate to regulate cumulus cell function. To identify potential mechanisms involved in these interactions, the ability of fully grown oocytes to regulate expression of BMP or FGF antagonists in cumulus cells was examined. Oocytes promoted cumulus cell expression of transcripts encoding antagonists to TGFbeta superfamily members, including Grem2, Htra1, Htra3, and Nog mRNAs. In contrast, oocytes suppressed cumulus cell expression of Spry2 mRNA, which encodes a regulator of receptor tyrosine kinase signals, such as FGF and epidermal growth factor (EGF) receptor signals. The regulation of Spry2 mRNA levels in cumulus cells was studied further as a model for analysis of potential mechanisms for cooperativity of FGF/EGF signaling with oocyte-derived members of the TGFbeta superfamily. Oocytes suppressed basal and FGF-stimulated Spry2 mRNA levels in cumulus cells but promoted EGF-stimulated levels. Furthermore, recombinant TGFbeta superfamily proteins, including BMP15 and GDF9, mimicked these effects of oocytes. Elevated expression of Spry2 mRNA in cumulus and mural granulosa cells correlated with human chorionic gonadotropin-induced expression of mRNAs encoding EGF-like peptides. Therefore, oocyte-derived members of the TGFbeta superfamily suppress FGF-stimulated Spry2 mRNA levels before the luteinizing hormone surge but promote Spry2 mRNA levels stimulated by EGF receptor-mediated signals after the surge. PMID- 19553597 TI - Evidence that a defective spindle assembly checkpoint is not the primary cause of maternal age-associated aneuploidy in mouse eggs. AB - Advanced maternal age is unequivocally associated with increased aneuploidy in human eggs and infertility, but the molecular basis for this phenomenon is unknown. An age-dependent deterioration of the spindle assembly checkpoint (SAC) has been proposed as a probable cause of aneuploidy. Accurate chromosome segregation depends on correct chromosome attachment to spindle microtubules, and the SAC provides time for this process by delaying anaphase onset until all chromosomes are stably attached. If SAC function decreases with age, oocytes from reproductively old mice would enter anaphase of meiosis I (AI) prematurely, leading to chromosome segregation errors and aneuploid eggs. Although intuitively appealing, this hypothesis is largely untested. We used a natural reproductive aging mouse model to determine if a defective SAC is the primary cause of aneuploidy in eggs. We tracked the progress of individual oocytes from young and old mice through meiosis I by time-lapse microscopy and counted chromosomes in the resulting eggs. This data set allowed us to correlate the timing of AI onset with aneuploidy in individual oocytes. We found that oocytes from old mice do not enter AI prematurely compared to young counterparts despite a 4-fold increase in the incidence of aneuploidy. Moreover, we did not observe a correlation between the timing of AI onset and aneuploidy in individual oocytes. When SAC function was challenged with a low concentration of the spindle toxin nocodazole, oocytes from both young and old mice arrested at meiosis I, which is indicative of a functional checkpoint. These findings indicate that a defective SAC is unlikely the primary cause of aneuploidy associated with maternal age. PMID- 19553598 TI - Overactive beta-catenin signaling causes testicular sertoli cell tumor development in the mouse. AB - Overactive WNT/beta-catenin signaling has been found in many forms of cancer in human patients. Mouse models with mutations in different components of the WNT/beta-catenin signaling pathway have been generated to mimic tumorigenesis in humans. Mice with mutations that result in overactive WNT/beta-catenin signaling developed tumors in some tissues, such as digestive tract, skin, and ovary, but they failed to develop tumors in other tissues, such as mammary gland, liver, kidney, and primordial germ cells. To investigate whether overactive beta-catenin signaling is capable of inducing Sertoli cell tumorigenesis in testes, we generated Ctnnb1(tm1Mmt/+);Tg(AMH-cre)1Flor male mice that express a constitutively active form of beta-catenin specifically in Sertoli cells. No tumors were observed at 4 mo of age, but 70% of the mutant males developed Sertoli cell tumors at 8 mo of age. At 1 yr of age, more than 90% of the mutant males developed tumors. No instances of extratesticular spread of the tumors were found in the mutant mice. These studies show a causal link between overactive WNT/beta-catenin signaling and Sertoli cell tumor development and provide a novel mouse model for the study of Sertoli cell tumor biology. PMID- 19553599 TI - Risk assessment of minute virus of mice transmission during rederivation: detection in reproductive organs, gametes, and embryos of mice after in vivo infection. AB - Murine parvoviruses, including minute virus of mice (MVM), represent major infectious disease problems encountered in contemporary laboratory animal research facilities with embryo transfer (ET), one of the most widely used techniques for rederivation. Using an in vivo approach, the objectives of this study were to assess the risk of MVM transmission during rederivation and to provide data that allow recommendation of preventive measures. Therefore, we determined whether immunosuppressive variant MVMi viral DNA is detectable in reproductive organs, gametes (oocytes and spermatozoa), and embryos collected from experimentally infected mice and whether washing as recommended before ET eliminates MVMi sufficiently from gametes and embryos. Fractions of reproductive organs tested positive from Day 5 to Day 30 postinoculation, demonstrating a risk for a minimum period of 4 wk; the highest incidence of positive organs was found between Day 9 and Day 13 postinoculation. Real-time PCR detected viral DNA to a lesser extent in male than in female reproductive organs. MVMi DNA was detected in oocytes and sperm cells derived after in vivo infection but not in two-cell embryos. In vitro contamination studies revealed that the virus firmly adheres to the zona pellucida after 10 wash steps, indicating that even extensive washing might not eliminate MVMi completely from embryos. According to this systematic in vivo approach, recommended measures to prevent transmission of MVM during rederivation include sufficient washing of embryos, accompanying testing using adequate (PCR) methods, and using embryos rather than in vitro fertilization techniques; furthermore, the exchange of gametes should be considered a risk factor. PMID- 19553600 TI - Platelet-derived growth factor C is upregulated in human uterine fibroids and regulates uterine smooth muscle cell growth. AB - Leiomyomata uteri (i.e., uterine fibroids) are benign tumors arising from the abnormal growth of uterine smooth muscle cells (SMCs). We show here that the expression of platelet-derived growth factor C (PDGFC) is higher in approximately 80% of uterine fibroids than in adjacent myometrial tissues examined. Increased expression of PDGFC is also observed in fibroid-derived SMCs (fSMCs) relative to myometrial-derived SMCs (mSMCs). Recombinant bioactive PDGFCC homodimer stimulates the growth of fSMCs and mSMCs in ex vivo cultures and prolongs the survival of fSMCs in Matrigel plugs implemented subcutaneously in immunocompromised mice. The knockdown of PDGF receptor-alpha (PDGFRA) through lentiviral-mediated RNA interference reduces the growth of fSMCs and mSMCs in ex vivo cultures and in Matrigel implants. Furthermore, two small molecule inhibitors of the PDGFR tyrosine kinase (i.e., imatinib and dasatinib) exerted negative effects on fSMC and mSMC growth in ex vivo cultures, albeit at concentrations that cannot be achieved in vivo. These results suggest that the PDGFCC/PDGFRA signaling module plays an important role in fSMC and mSMC growth, and that the upregulation of PDGFC expression may contribute to the clonal expansion of fSMCs in the development of uterine fibroids. PMID- 19553601 TI - Involvement of phospholipase C-related inactive protein in the mouse reproductive system through the regulation of gonadotropin levels. AB - Phospholipase C-related but catalytically inactive protein (comprising PRIP-1 and PRIP-2 [officially designated PLCL1 and PLCL2]) was first identified in our laboratory, but the biological functions have remained elusive. Therefore, we generated Plcl1 and Plcl2 double-knockout mice (Plcl1(tm1Mh); Plcl2(tm1Tta)) to gain insight into the biological function. Double-knockout mice apparently grew normally and became fertile; however, during animal maintenance, we noticed that mutant couples exhibited decreased litter events and litter size, indicating dysfunction of the reproductive system. Cross-mating experiments to discriminate whether males or females were defective indicated that the cause appeared to be on the female side. Mutant female mice had an apparently smaller uterus by gross anatomical observation and had more estrous days during the cycles. Levels of serum luteinizing hormone (LH) and follicle-stimulating hormone were measured for 5-6 consecutive days and were significantly higher in the mutant, which was also confirmed by examining the secretion of LH from the explant culture of anterior pituitary glands of wild-type and double-knockout mice. These results suggest that through gonadotropin secretion, PRIP plays an important role in female reproduction. PMID- 19553602 TI - Leptin stimulates protein synthesis-activating translation machinery in human trophoblastic cells. AB - Leptin was originally considered as an adipocyte-derived signaling molecule for the central control of metabolism. However, pleiotropic effects of leptin have been identified in reproduction and pregnancy, particularly in placenta, where it may work as an autocrine hormone, mediating angiogenesis, growth, and immunomodulation. Leptin receptor (LEPR, also known as Ob-R) shows sequence homology to members of the class I cytokine receptor (gp130) superfamily. In fact, leptin may function as a proinflammatory cytokine. We have previously found that leptin is a trophic and mitogenic factor for trophoblastic cells. In order to further investigate the mechanism by which leptin stimulates cell growth in JEG-3 cells and trophoblastic cells, we studied the phosphorylation state of different proteins of the initiation stage of translation and the total protein synthesis by [(3)H]leucine incorporation in JEG-3 cells. We have found that leptin dose-dependently stimulates the phosphorylation and activation of the translation initiation factor EIF4E as well as the phosphorylation of the EIF4E binding protein EIF4EBP1 (PHAS-I), which releases EIF4E to form active complexes. Moreover, leptin dose-dependently stimulates protein synthesis, and this effect can be partially prevented by blocking mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3 kinase (PIK3) pathways. In conclusion, leptin stimulates protein synthesis, at least in part activating the translation machinery, via the activation of MAPK and PIK3 pathways. PMID- 19553604 TI - Activation of a chloride channel by a trophic ligand is required for development of the mouse preimplantation embryo in vitro. AB - Platelet-activating factor (1-o-alkyl-2-acetyl-sn-gylcero-3-phosphocholine [PAF]) is one of several autocrine trophic factors supporting the development of the preimplantation embryo. PAF acts on the embryo to induce receptor-mediated intracellular calcium (Ca(2+))(i) transients, and these coincide with a marked membrane hyperpolarization. Patch-clamp analysis of 2-cell embryos showed that these Ca(2+)(i) transients resulted in an outward membrane current. The present study characterizes this current and assesses its role in embryo development. The outward current was dependent upon the presence of anions in the extracellular medium and occurred as a consequence of the PAF-induced Ca(2+)(i) transients. The anion current induced by PAF was inhibited by niflumic acid (NFA), a selective blocker of Ca(2+)-activated Cl(-) channels, but this drug did not block the PAF induced Ca(2+)(i) transients. Voltage ramp analysis showed that the Cl(-) conductance was outwardly rectifying and inactivated at holding potentials more positive than +30 mV. Culture in NFA or 4,4'-diisothiocyanatostilbene-2,2' disulfonic acid (a broad-specificity anion channel blocker) from the zygote stage significantly reduced development to blastocysts, with most arresting at the 4 cell and 8-cell stages. Niflumic acid exposure only from the zygote to the late 2 cell stage also reduced the subsequent development to blastocysts. By contrast, treatment from the late 2-cell stage or the 8-cell stage had no effect on development to the blastocyst stage. This study demonstrates the activation of a Ca(2+)-sensitive Cl(-) channel in the 2-cell embryo by PAF and shows that this current activity during the zygote to 2-cell stage is required for normal embryo development in vitro. PMID- 19553603 TI - Linker histones stimulate HSPA2 ATPase activity through NASP binding and inhibit CDC2/Cyclin B1 complex formation during meiosis in the mouse. AB - In mammalian spermatocytes, cell division cycle protein 2 (CDC2)/cyclin B1 and the chaperone heat shock protein A2 (HSPA2) are required for the G2-->M transition in prophase I. Here, we demonstrate that in primary spermatocytes, linker histone chaperone testis/embryo form of nuclear autoantigenic sperm protein (tNASP) binds the heat shock protein HSPA2, which localizes on the synaptonemal complex of spermatocytes. Significantly, the tNASP-HSPA2 complex binds linker histones and CDC2, forming a larger complex. We demonstrate that increasing amounts of tNASP favor tNASP-HSPA2-CDC2 complex formation. Binding of linker histones to tNASP significantly increases HSPA2 ATPase activity and the capacity of tNASP to bind HSPA2 and CDC2, precluding CDC2/cyclin B1 complex formation and, consequently, decreasing CDC2/cyclin B1 kinase activity. Linker histone binding to NASP controls the ability of HSPA2 to activate CDC2 for CDC2/cyclin B1 complex formation; therefore, tNASP's role is to provide the functional link between linker histones and cell cycle progression during meiosis. PMID- 19553605 TI - Progesterone-regulated changes in endometrial gene expression contribute to advanced conceptus development in cattle. AB - The postovulatory rise in circulating progesterone (P4) concentrations is associated with increased pregnancy success in beef and dairy cattle. Our study objective was to determine how elevated P4 alters endometrial gene expression to advance conceptus development. Synchronized heifers were inseminated (Day 0) and randomly assigned to pregnant high P4 or to pregnant normal P4. All high P4 groups received a P4-release intravaginal device on Day 3 after insemination that increased P4 concentrations up to Day 7 (P < 0.05). Tissue was collected on Day 5, 7, 13, or 16 of pregnancy, and endometrial gene expression was analyzed using the bovine Affymetrix (Santa Clara, CA) microarrays. Microarray analyses demonstrated that the largest number of P4-regulated genes coincided with the day when the P4 profiles were different for the longest period. Genes with the largest fold change increase (such as DGAT2 and MSTN [also known as GDF8]) were associated with triglyceride synthesis and glucose transport, which can be utilized as an energy source for the developing embryo. Temporal changes occurred at different stages of early pregnancy, with the greatest difference occurring between well-separated stages of conceptus development. Validation of a number of genes by quantitative real-time PCR indicated that P4 supplementation advances endometrial gene expression by altering the time (FABP, DGAT2, and MSTN) or duration (CRYGS) of expression pattern for genes that contribute to the composition of histotroph. PMID- 19553606 TI - Effects of diethylstilbestrol on programmed oocyte death and induction of polyovular follicles in neonatal mouse ovaries. AB - In mice, neonatal exposure to a synthetic estrogen, diethylstilbestrol (DES), induces polyovular follicles, which contain two or more oocytes per ovarian follicle. We reported previously that the estrogen receptor beta (ESR2) mediates DES signaling in polyovular follicle induction. However, the specific mechanism of polyovular follicle induction has not yet been clarified. Folliculogenesis in rodents begins soon after birth, accompanied by programmed oocyte death and germ cell loss. In this study, the effects of DES on oocyte death and on mRNA expression of genes thought to be involved in polyovular follicle induction were analyzed during a crucial period of folliculogenesis in the ovary of C57BL/6J, Fas(lpr/lpr) (lacking cell death receptor, FAS), and Esr2 knockout (Esr2 KO) mice. Neonatal DES exposure reduced programmed oocyte death in C57BL/6J mice; however, this reduction was not observed in Esr2 KO mice. In control Fas(lpr/lpr) mice, the oocyte apoptotic index was significantly lower than that in the control C57BL/6J mice. However, the polyovular follicle incidence in control 20-day-old Fas(lpr/lpr) mice was similar to that in the control C57BL/6J mice. Moreover, DES exposure changed mRNA expression of inhibin-alpha (Inha) in 2-day-old C57BL/6J mice. These results suggest that inhibition of oocyte death by DES through ESR2 may be one of the triggers for polyovular follicle induction. The FAS system is also involved in neonatal oocyte death; however, reduction of oocyte death is not sufficient for polyovular follicle induction. The combination of increased Inha mRNA and reduction of oocyte death in the ovaries of mice by DES through ESR2 might be correlated with polyovular follicle induction. PMID- 19553607 TI - Test-retest reliability of saccadic measures in subjects at risk for Huntington disease. AB - PURPOSE: Abnormalities in saccades appear to be sensitive and specific biomarkers in the prediagnostic stages of Huntington disease (HD). The goal of this study was to evaluate test-retest reliability of saccadic measures in prediagnostic carriers of the HD gene expansion (PDHD) and normal controls (NC). METHODS: The study sample included 9 PDHD and 12 NC who completed two study visits within an approximate 1-month interval. At the first visit, all participants completed a uniform clinical evaluation. A high-resolution, video-based system was used to record eye movements during completion of a battery of visually guided, antisaccade, and memory-guided tasks. Latency, velocity, gain, and percentage of errors were quantified. Test-retest reliability was estimated by calculating the intraclass correlation (ICC) of the saccade measures collected at the first and second visits. In addition, an equality test based on Fisher's z-transformation was used to evaluate the effects of group (PDHD and NC) and the subject's sex on ICC. RESULTS: The percentage of errors showed moderate to high reliability in the antisaccade and memory-guided tasks (ICC = 0.64-0.93). The latency of the saccades also demonstrated moderate to high reliability (ICC = 0.55-0.87) across all tasks. The velocity and gain of the saccades showed moderate reliability. The ICC was similar in the PDHD and NC groups. There was no significant effect of sex on the ICC. CONCLUSIONS: Good reliability of saccadic latency and percentage of errors in both antisaccade and memory-guided tasks suggests that these measures could serve as biomarkers to evaluate progression in HD. PMID- 19553608 TI - Development of microelectrode arrays for artificial retinal implants using liquid crystal polymers. AB - PURPOSE: To develop a liquid crystal polymer (LCP)-based, long-term implantable, retinal stimulation microelectrode array using a novel fabrication method. METHODS: The fabrication process used laser micromachining and customized thermal press bonding to produce LCP-based microelectrode arrays. To evaluate the fabrication process and the resultant electrode arrays, in vitro reliability tests and in vivo animal experiments were performed. The in vitro tests consisted of electrode site impedance recording and electrode interlayer adhesion monitoring during accelerated soak tests. For in vivo testing, the fabricated electrode arrays were implanted in the suprachoroidal space of rabbit eyes. Optical coherence tomography (OCT) and electrically evoked cortical potentials (EECPs) were used to determine long-term biocompatibility and functionality of the implant. RESULTS: The fabricated structure had a smooth, rounded edge profile and exhibited moderate flexibility, which are advantageous features for safe implantation without guide tools. After accelerated soak tests at 75 degrees C in phosphate-buffered saline, the electrode sites showed no degradation, and the interlayer adhesion of the structure showed acceptable stability for more than 2 months. The electrode arrays were safely implanted in the suprachoroidal space of rabbit eyes, and EECP waveforms were recorded. Over a 3-month postoperative period, no chorioretinal inflammation or structural deformities were observed by OCT and histologic examination. CONCLUSIONS: LCP-based flexible microelectrode arrays can be successfully applied as retinal prostheses. The results demonstrate that such electrode arrays are safe, biocompatible, and mechanically stable and that they can be effective as part of a chronic retinal implant system. PMID- 19553609 TI - Contribution of copy number variation in the regulation of complement activation locus to development of age-related macular degeneration. AB - PURPOSE: To develop an assay for determining the number of copies of the genes encoding complement factor H related 3 (CFHR3) and 1 (CFHR1) and determine the contribution of copy number variation (CNV) at CFHR3 and CFHR1 to the development of age-related macular degeneration (AMD). METHODS: A multiplex ligation dependent probe amplification (MLPA) assay was developed to quantify the number of copies of CFHR3 and CFHR1 in humans. Subjects with (n = 252) and without (n = 249) AMD were genotyped using the assay, and the impact on AMD risk was evaluated. RESULTS: The MLPA assay provided a consistent estimate of the number of copies of CFHR3 and CFHR1 in 500 of the 501 samples. Four different combinations of CNVs were observed with frequencies as follows: both CFHR3 and CFHR1 deletion (14%), CFHR3-only deletion (0.4%), CFHR1-only deletion (1.1%), and CFHR1 duplication (0.1%). Deletion of both copies of CFHR3 and CFHR1 decreased the odds of having AMD eightfold (95% CI 2-36) and always occurred on a protective haplotype, never on the risk haplotype tagged by the Y402H risk allele in CFH. The protection conferred by deletion of CFHR3 and CFHR1 could not be distinguished from the absence of the risk haplotype. CONCLUSIONS: Both deletions and duplications of genes in the regulation of complement activation locus segregated in Caucasians. Deletion of CFHR3 and CFHR1 protected against the development of AMD at least in part because the deletion tagged a protective haplotype and did not occur on the risk haplotype. PMID- 19553610 TI - Interobserver agreement for the detection of optical coherence tomography features of neovascular age-related macular degeneration. AB - PURPOSE: To investigate the interobserver agreement for the detection of optical coherence tomography (OCT) features of disease activity in patients with neovascular age-related macular degeneration (nAMD). METHODS: This was a cross sectional agreement study in which grading of OCT line scans from patients with nAMD was conducted by two retinal specialists before the patients received treatment. Scans were graded for the presence of features of nAMD disease activity (intraretinal cysts [IRC], subretinal fluid [SRF], diffuse retinal edema [DRE], retinal pigment epithelial detachment [PED], and subretinal tissue [SRT]). RESULTS: Although scans from 78 patients were available for analysis, five patients were excluded because of a mean signal strength of <7. Two hundred seventy-eight line scans were analyzed from 73 patients (40 with cross-hair scan sets and 33 with radial line scan sets). Agreement for per line scan analysis was 77% for IRC (kappa = 0.41), 81% for SRF (kappa = 0.62), 91% for macular fluid (kappa = 0.28), 79% for DRE (kappa = 0.10), 90% for PED (kappa = 0.78), and 79% for SRT (kappa = 0.53). Both observers disagreed regarding the presence of macular fluid in one patient (with a cross-hair scan). CONCLUSIONS: Interpretation of OCT line scans from patients with nAMD is subject to interobserver variability. However, when all line scans acquired are examined for the presence of fluid (IRC or SRF), there is a high level of agreement for the detection of macular fluid on a per patient basis. PMID- 19553611 TI - Infection of human retinal pigment epithelial cells with influenza A viruses. AB - PURPOSE: Ocular involvement in influenza A virus diseases is common but usually limited to mild conjunctivitis. Rarely, inflammation of the choriocapillaris may result in atrophia of the retinal pigment epithelium (RPE). Primary human retinal pigment epithelial (RPE) cells were infected with seasonal (H1N1 A/New Caledonia/20/99, H3N2 A/California/7/2004) or highly pathogenic avian H5N1 (A/Thailand/1(Kan-1)/04, A/Vietnam/1203/04, A/Vietnam/1194/04) influenza strains. METHODS: Influenza A virus replication was studied by investigation of cytopathogenic effects, immune staining for influenza A virus nucleoprotein, determination of virus titers, and electron microscopy. Apoptosis induction was examined by immune staining for activated caspase 3 and cleaved PARP. Proinflammatory gene expression was investigated by quantitative PCR. RESULTS: H5N1 but not seasonal influenza strains replicated to high titers (>10(8) TCID(50)/mL; 50% tissue culture infectious dose/milliliter) in RPE cells. H5N1 infection resulted in RPE cell apoptosis that was abolished by the antiviral drug ribavirin. Pretreatment with type I interferons (interferon-alpha and -beta) or the type II interferon, (interferon-gamma), inhibited H5N1 replication. Moreover, H5N1 infection induced expression of proinflammatory genes (tumor necrosis factor alpha, CXCL8, CXCL10, CXCL11, and interleukin-6), which was inhibited by ribavirin in a concentration-dependent manner. CONCLUSIONS: A novel cell type derived from the central nervous system was permissive to H5N1 influenza virus replication. This findings supports those suggesting H5N1 influenza strains to own a greater potential to spread to nonrespiratory tissues than seasonal human influenza viruses. Moreover, the data warrant the further study of the role of influenza A virus replication in retinal diseases associated with influenza A virus infections. PMID- 19553612 TI - Primary open-angle glaucoma: association with cholesterol 24S-hydroxylase (CYP46A1) gene polymorphism and plasma 24-hydroxycholesterol levels. AB - PURPOSE: Genetics has made significant contributions to the study of glaucoma over the past few decades. Cholesterol-24S-hydroxylase (CYP46A1) is a cholesterol metabolizing enzyme that is especially expressed in retinal ganglion cells. CYP46A1 and its metabolic product, 24S-hydroxycholesterol, have been linked to neurodegeneration. A single-nucleotide polymorphism (SNP) in the CYP46A1 gene, designated as rs754203 and associated with Alzheimer disease, was evaluated as a genetic risk factor for primary open-angle glaucoma (POAG), as well as plasma 24S hydroxycholesterol levels. METHODS: The frequency of the CYP46*C and CYP46*T alleles was analyzed in 150 patients with POAG and 118 control subjects. Plasma 24S-hydroxycholesterol levels were quantified. Sex, age, alleles, and genotype frequencies between patients with POAG and control subjects were compared by using the chi(2) and Student's t-tests. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by logistic regression to assess the relative association between disease and age, sex, and genotypes. RESULTS: The frequency of the TT genotype was significantly higher in patients with POAG than in control subjects (61.3% versus 48.3%, respectively, OR = 1.26; 95% CI = 1.006-1.574, P < 0.05). Plasma 24S-hydroxycholesterol levels did not differ between control subjects and patients with POAG. The ratio of estimated brain weight to liver volume as an estimate of the capacity of the human body to synthesize and metabolize 24S-hydroxycholesterol was found to correlate to plasma 24S hydroxycholesterol in control subjects and patients with POAG. CONCLUSIONS: The rs754203 SNP in CYP46A1 was associated with a risk for POAG. This polymorphism was not associated with changes in plasma 24S-hydroxycholesterol, highlighting that despite its retinal origin, 24S-hydroxycholesterol cannot be used as a biomarker for POAG. PMID- 19553614 TI - Topographic arrangement of S-cone photoreceptors in the retina of the diurnal Nile grass rat (Arvicanthis niloticus). AB - PURPOSE: The retina of Arvicanthis niloticus, a diurnal murine rodent closely related to Rattus (rats) and Mus (mice), contains approximately 30% to 35% cones and has several cone-driven functional characteristics found in humans. In this study the organization of these cone photoreceptors was examined, with emphasis on those expressing the S-opsin photopigment (S-cones). METHODS: Cones were labeled with antibodies against M- and S-opsins. Their topographic arrangement was examined on images of retinal flatmounts using density measures, nearest neighbor distance, and Voronoi domain analysis. Partial sequencing of the S-opsin DNA was also performed to determine whether this visual pigment was blue/violet or UV sensitive. RESULTS: Cone photoreceptors (estimated total population approximately 1.450 million) came in two distinct types that express either M/L- or S-opsin. Both types were present across the retinal surface. S-cones (approximately 7-8% of the total cone population) achieved a higher density in a discrete temporodorsal sector of the retina. The S-cone mosaic was irregular. Finally, S-cones were likely to be UV sensitive, according to genetic analysis. CONCLUSIONS: The topographic arrangement of cone photoreceptors in the retina of the diurnal Nile grass rat A. niloticus represents a highly pertinent model to improve understanding of the pathologic course of and related therapy for retinal disease involving cones. PMID- 19553613 TI - Role of alpha 4 integrin (CD49d) in the pathogenesis of diabetic retinopathy. AB - PURPOSE: The pathophysiology of diabetic retinopathy is mediated by leukocyte adhesion to the vascular endothelium of the diabetic retina, which results in endothelial injury, blood-retina barrier breakdown, and capillary nonperfusion. Leukocyte adhesion is triggered by the interaction of vascular endothelium adhesion molecules, such as ICAM-1, with leukocyte integrins, such as CD18. Inhibition of ICAM-1/CD18 signaling suppresses but does not completely abolish the cardinal manifestations of diabetic retinopathy, suggesting a role for additional adhesion molecules. Integrin alpha 4 (CD49d), in complex with integrin beta1, forms very late antigen-4 (VLA-4), which interacts with vascular cell adhesion molecule-1. The authors have now studied the role of integrin alpha 4/CD49d in the pathogenesis of diabetic retinopathy. METHODS: Diabetes mellitus was induced in Long Evans rats with streptozotocin, and an anti-alpha 4 integrin/CD49d neutralizing antibody was injected 5 and 10 days later. Two weeks after streptozotocin administration, vascular leakage was quantified with the Evans Blue technique. Leukostasis was measured with a static adhesion assay ex vivo and the FITC-lectin perfusion method in vivo. Retinal VEGF and TNF-alpha levels and NF-kappaB activity were measured by ELISA. RESULTS: Blockade of alpha 4 integrin/CD49d attenuated the diabetes-induced upregulation of NF-kappaB activation, VEGF, and TNF-alpha protein levels and reduced significantly diabetes induced leukocyte adhesion and vascular leakage. CONCLUSIONS: These data identify alpha 4 integrin/CD49d as a mediator of leukocyte adhesion and the resultant early signature abnormalities of diabetic retinopathy. Inhibition of this signaling pathway may hold promise for clinical activity in patients with diabetes. PMID- 19553615 TI - Corneal subbasal nerves changes in patients with diabetic retinopathy: an in vivo confocal study. AB - PURPOSE: To study the subbasal corneal plexus (SCP) in patients with diabetic retinopathy (DR) treated or nontreated with panretinal Argon laser photocoagulation (ALP). METHOD: Fifty consecutive patients with DR and 50 age- and sex-matched normal control subjects were examined with retinal tomography by a masked evaluator. The following subbasal plexus nerves parameters were considered: number per frame, tortuosity, and reflectivity. Diabetic patients were divided into two groups, according to the presence of proliferative versus nonproliferative retinopathy, according to the Early Treatment of Diabetic Retinopathy Study (ETDRS) classification. RESULTS: The number of fibers per frame and reflectivity were significantly lower in diabetic patients compared with control subjects (2.4 +/- 1 vs. 2.9 +/- 0.8, P = 0.01 and 2.3 +/- 0.9 vs. 2.6 +/- 0.9, P = 0.04, respectively). Tortuosity was significantly higher in diabetic patients (2.5 +/- 0.9 vs. 2.0 +/- 0.8, P = 0.002). Number per frame and reflectivity were significantly lower in diabetic patients with proliferative diabetic retinopathy (PDR; respectively, 2.0 +/- 0.9 vs. 2.9 +/- 0.9, P = 0.001, and 2.0 +/- 0.8 vs. 2.6 +/- 0.7, P = 0.003). Tortuosity was significantly higher in the PDR group (2.2 +/- 0.8 vs. 2.8 +/- 0.9, P = 0.008). The PDR group treated with ALP had significantly lower subbasal nerves number compared with the nontreated group (P = 0.01). CONCLUSIONS: DR may induce substantial changes in the SCP. There is a difference between proliferative and nonproliferative retinopathy and in the former group between ALP treated and nontreated patients. PMID- 19553616 TI - Regional differences in store-operated Ca2+ entry in the epithelium of the intact human lens. AB - PURPOSE: An elevated level of Ca(2+) is an important factor in cataract, yet precisely how Ca(2+) enters the lens is unknown. Lens epithelial cells contain a range of G-protein-coupled receptors and receptor tyrosine kinases that induce increases in intracellular Ca(2+). Receptor-associated Ca(2+) influx is, therefore, likely to be an important route for Ca(2+) influx to the lens. The authors investigated stimulated and passive Ca(2+) influx in in situ human lens epithelium. METHODS: Ca(2+) changes in equatorial (E) and central anterior (CA) epithelial cells were monitored with the use of a Ca(2+) indicator (Fluo4) and confocal microscopy. Gene expression was monitored by RT-PCR and immunoblotting. RESULTS: Adenosine triphosphate (ATP) induced Ca(2+) responses that were smaller in CA than E. Ca(2+) store depletion, using ATP (100 microM) or thapsigargin (1 microM), revealed greater relative store capacity and Ca(2+) influx in E. Ca(2+) influx was blocked by La(3+) (0.5 microM) in both regions. Unstimulated Ca(2+) influx was greater in E than CA. Greater expression of Orai1 and STIM1 was detected in E than in CA. CONCLUSIONS: Greater Ca(2+) store capacity and Ca(2+) influx in E compared with CA reflects underlying differences in proliferation and differentiation between the regions. The relatively small resting Ca(2+) influx in CA epithelium suggests that store-operated Ca(2+) entry (SOCE) is the main route of Ca(2+) influx in these cells. Greater resting influx and SOCE in E cells suggests that these are a major route for Ca(2+) influx into the lens. Increased expression of Orai1 and STIM1 in E could account for the differences in Ca(2+) entry. Receptor activation will modulate Ca(2+) influx, and inappropriate activity may contribute to cortical cataract. PMID- 19553618 TI - Refractive error and biometry in older Chinese adults: the Liwan eye study. AB - PURPOSE: To assess the prevalence of refractive error and describe the distribution of ocular biometry and its association with refraction in adult Chinese. METHODS: Random clustering sampling was used to identify adults aged > or = 50 years in Liwan District, Guangzhou. Refraction was determined by subjective refraction that achieved the best corrected vision based on monocular measurement. Ocular biometry was measured by A-mode ultrasound using a handheld applanation probe. RESULTS: Among 1405 participants in the study, data from 1269 phakic right eyes were available for analysis. The prevalence of myopia (SE < 0.5 D), hyperopia (SE > +0.5 D), and astigmatism (cylinder > 0.75 D) was 32.3% (95% confidence interval [CI], 27.8%-34.6%), 40.0% (95% CI, 37.3%-42.7%), and 48.3% (95% CI, 45.6%-51.1%), respectively. The spherical equivalent tended to become hyperopic at 60 years and shifted toward myopia at 75 years. Axial length did not change with age but was consistently shorter in women. Lens thickness increased with age and tended to be greater in women. CONCLUSIONS: The prevalence of myopia and biometric distribution in this urban Chinese cohort are similar to those observed in Singaporean Chinese but greater than in Mongolians and Europeans. Further studies are needed to clarify the role of environmental factors in the myopia rates. PMID- 19553617 TI - Differential effects of ADAMTS-1, -4, and -5 in the trabecular meshwork. AB - PURPOSE: Matrix metalloproteinases (MMPs) degrade extracellular matrix (ECM) and increase outflow facility in anterior segment perfusion culture. One group is the ADAMTSs (a disintegrin and metalloproteinase with thrombospondin type 1 motifs). In this study, the authors examined the effects of ADAMTS-1, -4, and -5 on outflow facility and investigated their mRNA levels and protein expression in the trabecular meshwork (TM). METHODS: ADAMTS mRNA was quantitated by qRT-PCR in TM cells exposed to TNFalpha, IL-1alpha, TGFbeta2, or mechanical stretch. ADAMTS-4 mRNA was assessed in normal and glaucomatous human anterior segments perfused at physiological or elevated pressure. Immunofluorescence was used to localize ADAMTSs in human TM cells and tissue. Anterior segments in perfusion culture were treated with recombinant ADAMTSs to determine effects on outflow facility. RESULTS: Cytokine treatment increased mRNA of all three ADAMTSs. Mechanical stretch increased ADAMTS-4 mRNA but conversely decreased ADAMTS-1 and -5 mRNA. ADAMTS-4 mRNA levels increased in response to pressure elevation in normal eyes and to higher levels in glaucomatous eyes. ADAMTS-4 protein was highly increased in the juxtacanalicular region of the TM in anterior segments perfused at increased pressure. In human TM cells, ADAMTS-4 colocalized with cortactin in podosome- or invadopodia-like structures, but ADAMTS-1 and -5 did not. Recombinant ADAMTS-4 increased outflow facility in human and porcine anterior segments, whereas recombinant ADAMTSs-1 and -5 did not. CONCLUSIONS: These results show differential responses and expression of ADAMTS-1, -4, and -5 in human TM cells. Combined, these results suggest that ADAMTS-4 is a potential modifier of outflow facility. PMID- 19553619 TI - Multimodal imaging including spectral domain OCT and confocal near infrared reflectance for characterization of outer retinal pathology in pseudoxanthoma elasticum. AB - PURPOSE: To investigate the value of multimodal confocal scanning laser ophthalmoscopy (cSLO) for phenotyping fundus lesions in patients with pseudoxanthoma elasticum (PXE) and to correlate these findings with morphologic alterations detected by spectral domain optical coherence tomography (SD-OCT). METHODS: Imaging was performed with a combined SD-OCT-cSLO system (Spectralis HRA OCT; Heidelberg Engineering, Heidelberg, Germany). OCT scans were placed at locations of interest on near-infrared (NIR) reflectance, fundus autofluorescence (FAF), and fluorescein angiography (FA) images. The instrument allowed for exact topographic correlation of findings on OCT and cSLO images. RESULTS: NIR reflectance imaging showed the highest sensitivity to detect angioid streaks and peau d'orange compared to FAF or FA. On OCT scans, angioid streaks reliably showed breaks in Bruch's membrane. Peau d'orange was associated with alternating reflectivity within Bruch's membrane. Characteristic mid-peripheral chorioretinal atrophies showed hyporeflective spaces involving the outer neurosensory retina. In eyes with pattern dystrophy like alterations, subneurosensory accumulation of material was observed within areas of increased FAF. CONCLUSIONS: SD-OCT in combination with cSLO imaging using NIR light locates the primary pathologic formations of angioid streaks and peau d'orange in Bruch's membrane. NIR reflectance imaging may be superior for detecting PXE-related fundus lesions at the level of Bruch's membrane, because the blue laser light that is used in FAF and FA is markedly absorbed by the pigment epithelium and therefore may only detect alterations if this cell layer is also affected. The findings indicate that multimodal cSLO and SD-OCT imaging of the outer retina allows for screening of PXE related retinal alterations. PMID- 19553620 TI - Impaired temporal, not just spatial, resolution in amblyopia. AB - PURPOSE: In amblyopia, neuronal deficits deteriorate spatial vision including visual acuity, possibly because of a lack of use-dependent fine-tuning of afferents to the visual cortex during infancy; but temporal processing may deteriorate as well. METHODS: Temporal, rather than spatial, resolution was investigated in patients with amblyopia by means of a task based on time-defined figure-ground segregation. Patients had to indicate the quadrant of the visual field where a purely time-defined square appeared. RESULTS: The results showed a clear decrease in temporal resolution of patients' amblyopic eyes compared with the dominant eyes in this task. The extent of this decrease in figure-ground segregation based on time of motion onset only loosely correlated with the decrease in spatial resolution and spanned a smaller range than did the spatial loss. Control experiments with artificially induced blur in normal observers confirmed that the decrease in temporal resolution was not simply due to the acuity loss. CONCLUSIONS: Amblyopia not only decreases spatial resolution, but also temporal factors such as time-based figure-ground segregation, even at high stimulus contrasts. This finding suggests that the realm of neuronal processes that may be disturbed in amblyopia is larger than originally thought. PMID- 19553621 TI - Blocking mast cell-mediated type I hypersensitivity in experimental allergic conjunctivitis by monocyte chemoattractant protein-1/CCR2. AB - PURPOSE: To characterize the roles played by monocyte chemoattractant protein-1 and its preferential receptor CCR2 (MCP-1/CCL2) in acute allergic inflammation. METHODS: The direct effects of MCP-1 were evaluated histologically after a subconjunctival injection of recombinant MCP-1 into naive mice. The mice were sensitized to ragweed pollen, and allergic conjunctivitis was induced by an allergen challenge. The location of the induced MCP-1 was determined by immunohistochemistry. Anti-MCP-1 antibody and CCR2-specific antagonist, RS 504393, were used to determine whether an inhibition of MCP-1 or CCR2 signals would suppress the allergen-induced immediate hypersensitivity reaction. The effect of blocking CCR2 was tested in vitro with isolated mast cells from connective tissue, to evaluate the co-stimulatory signals mediated by CCR2 in mast cells directly. RESULTS: A subconjunctival injection of MCP-1 stimulated conjunctival mast cell degranulation and recruited monocytes/macrophages. In the allergic conjunctivitis model, the allergen-induced MCP-1 protein was located in the monocytes/macrophages in the substantia propria of the conjunctiva. Blocking MCP-1 significantly suppressed the allergen-induced clinical signs and mast cell degranulation without affecting the allergen-specific IgE, or the release of Th2 cytokine from the isolated draining lymph node cells. Inhibition of CCR2 similarly suppressed the acute inflammatory responses. Consistent with the outcome of the disease model, inhibition of CCR2 suppressed allergen-specific degranulation of IgE-primed, isolated conjunctival mast cells. CONCLUSIONS: Stimulation of the co-stimulatory axis of CCR2 by MCP-1 is essentially required for mast cell-mediated hypersensitivity reactions in mouse eyes. PMID- 19553622 TI - Molecular characterization of human retinal progenitor cells. AB - PURPOSE: To examine the molecular profile of fetal human retinal progenitor cells (hRPCs) expanded in vitro and those grown in a co-culture system with mouse retina through the analysis of protein and gene expression and neurotransmitter stimulated calcium dynamics. METHODS: hRPCS were isolated from human retina of 14 to 18 weeks gestational age (GA) and expanded in vitro. Immunoblot, microarray, and immunocytochemistry (ICC) assays were performed on undifferentiated hRPCs and those co-cultured with mouse retinas for 2 weeks. Cell function was assessed by using calcium imaging. RESULTS: The ICC results showed a gradual decrease in the percentages of KI67-, SOX2-, and vimentin-positive cells from passages (P) 1 to P6, whereas a sustained expression of nestin and PAX6 was observed through P6. Microarray analysis of P1 hRPCs showed the expression of early retinal developmental genes: VIM (vimentin), KI67, NES (nestin), PAX6, SOX2, HES5, GNL3, OTX2, DACH1, SIX6, and CHX10 (VSX2). At P6, hRPCs continued to express VIM, KI67, NES, PAX6, SOX2, GNL3, and SIX6. On co-culture, there was a significant increase in the expression of MKI67, PAX6, SOX2, GNL3, SIX3, and RHO (rhodopsin). Calcium imaging showed a functional response to excitatory neurotransmitters. CONCLUSIONS: Fetal-derived hRPCs show molecular characteristics indicative of a retinal progenitor state up to P6 (latest passage studied). They show a progressive decrease in the expression of immature markers as they reach P6. These cells are functional, respond to excitatory neurotransmitters, and exhibit changes in expression patterns in response to co-culture with mouse retina. PMID- 19553623 TI - Limited roles of Rdh8, Rdh12, and Abca4 in all-trans-retinal clearance in mouse retina. AB - PURPOSE: Although the retinoid cycle is essential for vision, all-trans-retinal and the side products of this cycle are toxic. Delayed clearance of all-trans retinal causes accumulation of its condensation products, A2E, and all-trans retinal dimer (RALdi), both associated with human macular degeneration. The protective roles were examined of the all-trans-RDHs, Rdh8 and Rdh12, and the ATP binding cassette transporter Abca4, retinoid cycle enzymes involved in all-trans retinal clearance. METHODS: Mice genetically engineered to lack Rdh8, Rdh12, and Abca4, either singly or in various combinations, were investigated because all trans-retinal clearance is achieved by all-trans-RDHs and Abca4. Knockout mice were evaluated by spectral-domain optical coherence tomography (SD-OCT), electroretinography, retinal morphology, and visual retinoid profiling with HPLC and MS. ARPE19 cells were examined to evaluate A2E and RALdi oxidation and toxicity induced by exposure to UV and blue light. RESULTS: Rdh8(-/-)Abca4(-/-) and Rdh8(-/-)Rdh12(-/-)Abca4(-/-) mice displayed slowly progressive, severe retinal degeneration under room light conditions. Intense light-induced acute retinal degeneration was detected by SD-OCT in Rdh8(-/-)Rdh12(-/-)Abca4(-/-) mice. Amounts of A2E in the RPE correlated with diminished all-trans-retinal clearance, and the highest A2E amounts were found in Rdh8(-/-)Rdh12(-/-)Abca4(-/ ) mice. However oxidized A2E was not found in any of these mice, and A2E oxidation was not induced by blue light and UV illumination of A2E-loaded ARPE19 cells. Of interest, addition of all-trans-retinal did activate retinoic acid receptors in cultured cells. CONCLUSIONS: Rdh8, Rdh12, and Abca4 all protect the retina and reduce A2E production by facilitating all-trans-retinal clearance. Delayed all-trans-retinal clearance contributes more than A2E oxidation to light induced cellular toxicity. PMID- 19553625 TI - Anterior chamber angle opening during corneoscleral indentation: the mechanism of whole eye globe deformation and the importance of the limbus. AB - PURPOSE: To determine how mechanical interaction among iris, cornea, limbus, sclera, and IOP contribute to angle opening during indentation of the cornea or sclera. METHODS: A finite-element model of the globe was developed. The model consisted of three elastic isotropic segments--iris, cornea, and sclera--and a two-component anisotropic segment representing the limbus. The model was tested against published in vitro experiments and then applied to angle opening during indentation in vivo. Indentation of the central cornea with a cotton bud, indentation with a small or large eyecup during ultrasound biomicroscopy, indentation with a gonioscopy lens, and scleral indentation during goniosynechialysis were modeled. RESULTS: The anisotropic limbus model matched published data better than any isotropic model. Simulation of all clinical cases gave results in agreement with published observations. The model predicted angle opening during indentation by a cotton bud or small eyecup but angle narrowing when the sclera was indented by a large eyecup. The model of indentation gonioscopy showed narrowing of the angle on the indentation side and opening of the angle on the opposite side. Nonuniform opening of the angle was predicted when the scleral surface was indented. CONCLUSIONS: The two-component model of the stiff fibers embedded in a soft matrix captured the mechanical properties of the complex limbal region effectively. The success of this model suggests that, at least in part, corneoscleral mechanics drive angle opening rather than aqueous humor pressurization. PMID- 19553624 TI - Abnormal retinal vascular oxygen tension response to light flicker in diabetic rats. AB - PURPOSE: To test the hypothesis that the intravascular oxygen response to light flicker is abnormal in diabetes. METHODS: Ten eyes of normal rats and 10 eyes of rats made diabetic with streptozotocin were examined. Oxygen tension (PO(2)) was measured noninvasively in the retinal arteries and veins on optical section retinal images. PO(2) was estimated based on the quenching by oxygen of the phosphorescence of an intravenously injected palladium porphyrin molecular probe. Measurements were conducted with and without light flicker at 10 Hz. Oxygen saturation (SO(2)) was calculated with adjustment for the arterial pH. RESULTS: In the normal rats flicker induced an increase in arterial PO(2) and in the difference in arterial and venous (A-V difference) PO(2) from 51 +/- 5 (mean and SD) to 55 +/- 7 mm Hg and from 22 +/- 3 to 26 +/- 5 mm Hg, respectively (P < 0.002 and 0.015, respectively). Flicker induced an increase of arterial SO(2) and A-V SO(2) difference from 64% +/- 8% to 68% +/- 7% and from 34% +/- 4% to 38% +/- 6%, respectively (P < 0.002 and 0.035, respectively). No changes in PO(2) or SO(2) were observed with flicker in the veins. In the diabetic rats, no significant flicker-induced changes were seen in PO(2) or SO(2) in the retinal arteries, veins, or A-V differences. CONCLUSIONS: The diabetic rats lacked the flicker induced increase in arterial PO(2) and SO(2) and also the A-V difference in PO(2) and SO(2) observed in the normal rats. The best explanation appeared to be that diabetes impairs the increase in oxygen consumption normally provoked by light flicker. PMID- 19553626 TI - Common sequence variation in the VEGFA gene predicts risk of diabetic retinopathy. AB - PURPOSE: Vascular endothelial growth factor (VEGF) is a multifunctional cytokine that plays a role in angiogenesis and microvascular permeability. This study was conducted to determine whether common sequence variation in the VEGFA gene plays a role in the development of diabetic retinopathy (DR). METHOD: Five hundred fifty-four subjects with diabetes mellitus (DM) including 190 type 1 DM (T1DM) and 364 type 2 DM (T2DM) were recruited. The study group consisted of 235 participants without DR, 158 with nonproliferative DR (NPDR), 132 with proliferative DR (PDR), and 93 with clinically significant macular edema (CSME). Blinding DR was defined as severe NPDR, PDR, or CSME. Fifteen VEGFA tag single nucleotide polymorphisms (SNPs) were genotyped in all subjects and tested for association with blinding DR. RESULTS: Multiple tag SNPs in the VEGFA gene were associated with blinding DR. After controlling for sex, HbA1c, and duration of disease, in T1DM, the AA genotype of rs699946 (P = 0.007, odds ratio [OR], 4.1; 95% confidence interval [CI], 1.5-11.4) and the GG genotype of rs833068 (P = 0.017, OR, 3.1; 95% CI, 1.3-7.2) were most significantly associated. In T2DM, the C allele of rs3025021 (P = 0.002; OR, 3.8; 95% CI, 1.5-10.0) and the G allele of rs10434 (P = 0.002; OR, 2.6; 95% CI, 1.3-5.3) were most significantly associated with blinding DR. Haplotype analyses suggested an important role for the haplotype TCCGCG in blinding DR (P = 0.0004). CONCLUSIONS: Sequence variation in the VEGFA gene is associated with risk of developing blinding DR in T1DM and T2DM. Identifying specific genetic markers will allow for refined screening algorithms and earlier intervention in patients at highest risk. PMID- 19553628 TI - Pigment epithelium-derived factor (PEDF) attenuated capsaicin-induced neurotrophic keratouveitis. AB - PURPOSE: To reveal the influence of retrobulbar capsaicin treatment on rats' eyes and to test the protective effects of PEDF, a known neurotrophic and antiangiogenic substance, against neurotrophic keratouveitis. METHODS: A single retrobulbar injection of capsaicin (50 mg/kg) was performed in young rats, and the effect of subsequent retrobulbar injections of PEDF 3.2 or 6.4 microg was recorded. Tear fluid alterations were evaluated with the Schirmer test and corneal alterations with slit lamp biomicroscopy. Histopathologic alterations were studied with light and electron microscopy. The number of leukocytes (myeloid cells) in the anterior and posterior chambers, peripheral retina, and vitreous were quantitatively evaluated. RESULTS: Reduced tear secretion was found in capsaicin-treated rats compared with the control, but this effect was significantly attenuated by PEDF. Corneal ulceration developed and was followed by scar formation and neovascularization in the capsaicin-treated, and it was also significantly attenuated by PEDF treatment. Leukocyte infiltration of the anterior and posterior chambers, as well as the peripheral retina and vitreous, was also observed in capsaicin-treated eyes and was significantly reduced by PEDF treatment. The protective effects of PEDF were dose dependent for each parameter, even if the treatment was initiated at day 14 after the challenge. CONCLUSIONS: PEDF accelerated the recovery of tear secretion and also prevented capsaicin induced neurotrophic keratouveitis and peripheral vitreoretinal inflammation. These effects of PEDF, described herein for the first time, may have a clinical application in inflammatory and neovascular diseases of the eye. PMID- 19553627 TI - Ultrastructural and molecular biologic comparison of classic proprioceptors and palisade endings in sheep extraocular muscles. AB - PURPOSE: To analyze and compare the structural and molecular features of classic proprioceptors like muscle spindles and Golgi tendon organs (GTOs) and putative proprioceptors (palisade endings) in sheep extraocular muscle (EOMs). METHODS: The EOMs of four sheep were analyzed. Frozen sections or wholemount preparations of the samples were immunohistochemically labeled and analyzed by confocal laser scanning microscopy. Triple labeling with different combinations of antibodies against neurofilament, synaptophysin, and choline acetyltransferase (ChAT), as well as alpha-bungarotoxin and phalloidin, was performed. Microscopic anatomy of the nerve end organs was analyzed by transmission electron microscopy. RESULTS: The microscopic anatomy demonstrated that muscle spindles and GTOs had a perineural capsule and palisade endings a connective tissue capsule. Sensory nerve terminals in muscle spindles and GTOs contained only a few vesicles, whereas palisade nerve terminals were full of clear vesicles. Likewise, motor terminals in the muscle spindles' polar regions were full of clear vesicles. Immunohistochemistry showed that sensory nerve fibers as well as their sensory nerve terminals in muscle spindles and GTOs were ChAT-negative. Palisade endings were supplied by ChAT-positive nerve fibers, and the palisade complexes including palisade nerve terminals were also ChAT-immunoreactive. Motor terminals in muscle spindles were ChAT and alpha-bungarotoxin positive. CONCLUSIONS: The present study demonstrated in sheep EOMs that palisade endings are innervated by cholinergic axons exhibiting characteristics typical of motoneurons, whereas muscle spindles (except the polar regions) and GTOs are supplied by noncholinergic axons. These results raise the question of whether palisade endings are candidates for proprioceptors in EOMs. PMID- 19553630 TI - Herpes zoster pathogenesis and cell-mediated immunity and immunosenescence. AB - Herpes zoster, or shingles, is a localized disease characterized by unilateral radicular pain and a vesicular rash limited to the area of skin innervated by a single dorsal root or cranial sensory ganglion. Whereas varicella, or chickenpox, results from primary exogenous varicella-zoster virus (VZV) infection, herpes zoster is caused by reactivation of endogenous VZV that has persisted in latent form within sensory ganglia following an earlier episode of chickenpox. In contrast to recurrent herpes simplex, herpes zoster is commonly associated with severe pain: prodromal pain often precedes the rash by several days; pain usually accompanies the dermatomal rash of herpes zoster; and clinically significant pain and allodynia may persist for weeks, months, or even years after the herpes zoster rash has healed, a debilitating complication known as postherpetic neuralgia (PHN). The incidence and severity of herpes zoster and PHN increase with age in association with an age-related decline in cell-mediated immunity to VZV. The Shingles Prevention Study-a randomized double-blinded placebo-controlled trial-sought to evaluate the capacity of a live attenuated VZV vaccine to protect older adults from herpes zoster and PHN by boosting their waning cell-mediated immunity to VZV. The study demonstrated that the zoster vaccine produced significant reductions in the incidence of herpes zoster, in the burden of illness caused by herpes zoster, and in the incidence of PHN. PMID- 19553629 TI - Inhibition of chemokine receptor expression on uveal melanomas by CXCR4 siRNA and its effect on uveal melanoma liver metastases. AB - PURPOSE: To determine whether blocking the expression of the chemokine receptor CXCR4 using siRNA inhibits chemotactic responses of human uveal melanoma cells to liver-derived factors and prevents liver metastases. METHODS: Human uveal melanoma cells were transfected with CXCR4 siRNA or control siRNA and tested in vitro for chemotactic and invasive behavior in response to soluble factors produced by human liver cells. The effect of CXCR4 siRNA transfection on the formation of liver metastases was tested by injecting transfected melanoma cells into the spleen capsules of NOD-SCID mice, and metastases were quantified by measuring the human housekeeping gene hHPRT in livers. RESULTS: Blocking CXCR4 interaction with its ligand using anti-CXCL12 antibody resulted in a significant reduction in the chemotactic responses of uveal melanoma cells to soluble factors produced by human liver cells. Similarly, blocking CXCR4 gene expression by transfection with CXCR4 siRNA inhibited both the chemotactic and the invasive properties of uveal melanoma cells exposed to factors produced by human livers. Uveal melanoma cells transfected with CXCR4 siRNA produced fewer liver metastases than untreated uveal melanoma cells or uveal melanoma cells transfected with control siRNA. CONCLUSIONS: CXCR4 is a key chemokine receptor that may account for the organ-specific homing of human uveal melanomas to the liver, which contains significant quantities of CXCL2, the only known ligand for CXCR4. CXCR4 is a potential therapeutic target for preventing the initial establishment of liver metastases but has limited application for use in advanced liver tumors. PMID- 19553631 TI - Reducing the incidence and severity of herpes zoster and PHN with zoster vaccination. AB - The results of the Shingles Prevention Study, a Cooperative Studies Program conducted through the US Department of Veterans Affairs, are summarized. Also provided are general recommendations and contraindications for the use of zoster vaccine live among patients aged 60 years or older. PMID- 19553632 TI - Herpes zoster overview: natural history and incidence. AB - The varicella-zoster virus (VZV) causes two diseases. The primary VZV infection, known as chickenpox, typically occurs during childhood. Herpes zoster infection results later in life from reactivation of VZV in the dorsal root ganglia. Herpes zoster characteristically results in a rash with a unilateral dermatomal distribution, which usually resolves within 2 to 4 weeks. If the infection does not resolve after its acute phase, long-term complications, such as postherpetic neuralgia, may develop. The author discusses the natural history and incidence of primary VZV infection and herpes zoster and details the epidemiology, clinical manifestation, diagnosis, and complications of this disease. PMID- 19553633 TI - Herpes zoster vaccination: benefits and barriers. AB - The incidence and severity of herpes zoster increases with advancing age, as does its most common complication-postherpetic neuralgia (PHN). The most effective management strategy for herpes zoster and PHN is prevention of the disease through vaccination, particularly in the most vulnerable patient population (ie, individuals aged 70 years or older). The Shingles Prevention Study demonstrated that vaccination reduced the incidence and severity of herpes zoster, as well as the incidence of PHN. Despite the benefits of zoster immunization, administration of the vaccine may be met by certain barriers, including issues related to reimbursement, storage, and availability of the vaccine. Educating physicians and the public about the importance of herpes zoster prevention, while underscoring the pain associated with this disease and the challenges in managing it, will help to overcome these barriers. PMID- 19553634 TI - Overcoming barriers to adult immunization. AB - The herpes zoster vaccine has proven to reduce the incidence of herpes zoster and postherpetic neuralgia, a debilitating condition that can remain for years after zoster rash has resolved. However, few eligible adults have received the vaccine. Findings from survey-based studies on patient uptake of influenza and pneumococcal polysaccharide vaccines suggest that physician recommendations and standing orders can increase vaccine uptake. However, certain barriers unique to zoster vaccination (eg, reimbursement concerns) will need to be addressed. PMID- 19553635 TI - Managing herpes zoster and postherpetic neuralgia. AB - Treatment strategies for herpes zoster infection include limiting viral replication with anti-infective agents as well as limiting associated acute and chronic neuropathic pain with a variety of analgesics. The author outlines currently available pharmacotherapeutic options, from first-line (eg, anti infective agents, tricyclic antidepressants, anticonvulsants, opioid analgesics, and topical agents) through adjuvant treatments (eg, oral corticosteroids). A summary of adverse event profiles is provided for each medication. Vaccination is recommended as a preventive measure. PMID- 19553636 TI - Association of hereditary thrombocythemia and distal limb defects with a thrombopoietin gene mutation. AB - Hereditary thrombocythemia is a rare autosomal dominant disorder caused by mutations in either the thrombopoietin gene (TPO) or its receptor c-MPL. TPO mutations described so far lead to thrombopoietin overproduction through increased translation of m-RNA. Unilateral transverse reduction limb defects are usually sporadic and generally thought to be caused by vascular disruptions. Reports of inherited unilateral limb defects are extremely rare. In the present study, we describe a family with segregation of G185T TPO mutation in the 5' UTR region in 4 subjects with thrombocythemia. Three of these patients also present congenital transverse limb defects. Association of these events gives a strong hint of the in vivo involvement of thrombopoietin in vasculogenesis, confirming the role of TPO in human development of the hemangioblast, the embryonic progenitor of the hematopoietic and endothelial lineages. This is the first report showing that vascular disruptions could be secondary to specific gene derangements. PMID- 19553637 TI - TLR7 stimulation in human plasmacytoid dendritic cells leads to the induction of early IFN-inducible genes in the absence of type I IFN. AB - On recognition of influenza virus (Flu) by TLR7, plasmacytoid dendritic cells (pDCs) produce type I IFN in significant amounts. Synthetic TLR7 ligands induce the maturation of pDCs, as evidenced by the expression of costimulatory molecules and the production of proinflammatory cytokines; however, they induce only low level production of IFN-alpha. To dissect the TLR7 signaling in pDCs and how these different profiles are induced, we studied the effects of 2 TLR7 ligands (Flu and CL097) on the activation of blood-isolated pDCs and the human GEN2.2 pDC cell line. Type I IFN production by pDCs correlates with differential interferon regulatory factor 7 (IRF7) translocation into the nucleus induced by the 2 TLR7 ligands. Surprisingly, with both activators we nevertheless observed the rapid expression of the IFN-inducible genes mxa, cxcl10, and trail within 4 hours of stimulation. This expression, controlled by STAT1 phosphorylation, was independent of type I IFN. STAT1 activation was found to be strictly dependent on the PI3K-p38MAPK pathway, showing a new signaling pathway leading to rapid expression of IFN-inducible genes after TLR7 triggering. Thus, pDCs, through this unusual TLR7 signaling, have the capacity to promptly respond to viral infection during the early phases of the innate immune response. PMID- 19553638 TI - Improving survival in patients with chronic lymphocytic leukemia (1980-2008): the Hospital Clinic of Barcelona experience. AB - Whether advances in treatment are prolonging survival of patients with chronic lymphocytic leukemia (CLL) is unclear. We analyzed presentation patterns and survival over time in 929 patients followed from 1980 to 2008 at the Hospital Clinic of Barcelona. The 5- and 10-year relative survival (adjusted for the expected survival in the general population) was estimated in patients seen in 2 periods of time: 1980-1994 (n = 451) and 1995-2004 (n = 365). We found that CLL shortens life expectancy in all age groups independently of clinical features at diagnosis. Nevertheless, survival is improving, particularly in some groups of patients. Thus, relative survival was significantly higher in the 1995-2004 cohort than in the 1980-1994 group both at 5 years (incidence rate ratio [IRR] = 0.46; P = .004) and 10 years (IRR = 0.65; P = .007) from diagnosis. The improved survival was largely due to a decrease in CLL-attributable mortality in patients younger than 70 years in Binet stage B or C at diagnosis (IRR = 0.40; P = .001 at 5 years; IRR = 0.33; P < .001 at 10 years). These results suggest that newer treatments are changing the prognosis of CLL, particularly in younger patients with advanced disease, whereas no improvement is yet observed in older subjects or those with lower-risk disease. PMID- 19553639 TI - Preclinical characterization of 1-7F9, a novel human anti-KIR receptor therapeutic antibody that augments natural killer-mediated killing of tumor cells. AB - Inhibitory-cell killer immunoglobulin-like receptors (KIR) negatively regulate natural killer (NK) cell-mediated killing of HLA class I-expressing tumors. Lack of KIR-HLA class I interactions has been associated with potent NK-mediated antitumor efficacy and increased survival in acute myeloid leukemia (AML) patients upon haploidentical stem cell transplantation from KIR-mismatched donors. To exploit this pathway pharmacologically, we generated a fully human monoclonal antibody, 1-7F9, which cross-reacts with KIR2DL1, -2, and -3 receptors, and prevents their inhibitory signaling. The 1-7F9 monoclonal antibody augmented NK cell-mediated lysis of HLA-C-expressing tumor cells, including autologous AML blasts, but did not induce killing of normal peripheral blood mononuclear cells, suggesting a therapeutic window for preferential enhancement of NK-cell cytotoxicity against malignant target cells. Administration of 1-7F9 to KIR2DL3-transgenic mice resulted in dose-dependent rejection of HLA-Cw3 positive target cells. In an immunodeficient mouse model in which inoculation of human NK cells alone was unable to protect against lethal, autologous AML, preadministration of 1-7F9 resulted in long-term survival. These data show that 1 7F9 confers specific, stable blockade of KIR, boosting NK-mediated killing of HLA matched AML blasts in vitro and in vivo, providing a preclinical basis for initiating phase 1 clinical trials with this candidate therapeutic antibody. PMID- 19553640 TI - Synthetic lethality--a new direction in cancer-drug development. PMID- 19553641 TI - Inhibition of poly(ADP-ribose) polymerase in tumors from BRCA mutation carriers. AB - BACKGROUND: The inhibition of poly(adenosine diphosphate [ADP]-ribose) polymerase (PARP) is a potential synthetic lethal therapeutic strategy for the treatment of cancers with specific DNA-repair defects, including those arising in carriers of a BRCA1 or BRCA2 mutation. We conducted a clinical evaluation in humans of olaparib (AZD2281), a novel, potent, orally active PARP inhibitor. METHODS: This was a phase 1 trial that included the analysis of pharmacokinetic and pharmacodynamic characteristics of olaparib. Selection was aimed at having a study population enriched in carriers of a BRCA1 or BRCA2 mutation. RESULTS: We enrolled and treated 60 patients; 22 were carriers of a BRCA1 or BRCA2 mutation and 1 had a strong family history of BRCA-associated cancer but declined to undergo mutational testing. The olaparib dose and schedule were increased from 10 mg daily for 2 of every 3 weeks to 600 mg twice daily continuously. Reversible dose-limiting toxicity was seen in one of eight patients receiving 400 mg twice daily (grade 3 mood alteration and fatigue) and two of five patients receiving 600 mg twice daily (grade 4 thrombocytopenia and grade 3 somnolence). This led us to enroll another cohort, consisting only of carriers of a BRCA1 or BRCA2 mutation, to receive olaparib at a dose of 200 mg twice daily. Other adverse effects included mild gastrointestinal symptoms. There was no obvious increase in adverse effects seen in the mutation carriers. Pharmacokinetic data indicated rapid absorption and elimination; pharmacodynamic studies confirmed PARP inhibition in surrogate samples (of peripheral-blood mononuclear cells and plucked eyebrow-hair follicles) and tumor tissue. Objective antitumor activity was reported only in mutation carriers, all of whom had ovarian, breast, or prostate cancer and had received multiple treatment regimens. CONCLUSIONS: Olaparib has few of the adverse effects of conventional chemotherapy, inhibits PARP, and has antitumor activity in cancer associated with the BRCA1 or BRCA2 mutation. (ClinicalTrials.gov number, NCT00516373.) PMID- 19553642 TI - Images in clinical medicine. Lipomatosis of the tongue. PMID- 19553643 TI - A lifeline for primary care. PMID- 19553644 TI - Easing the shortage in adult primary care--is it all about money? PMID- 19553645 TI - A new era of for-profit hospice care--the Medicare benefit. PMID- 19553648 TI - Clinical practice. Alpha1-antitrypsin deficiency. PMID- 19553649 TI - Alcoholic hepatitis. PMID- 19553650 TI - Images in clinical medicine. Ingested magnets. PMID- 19553647 TI - Treating childhood acute lymphoblastic leukemia without cranial irradiation. AB - BACKGROUND: Prophylactic cranial irradiation has been a standard treatment in children with acute lymphoblastic leukemia (ALL) who are at high risk for central nervous system (CNS) relapse. METHODS: We conducted a clinical trial to test whether prophylactic cranial irradiation could be omitted from treatment in all children with newly diagnosed ALL. A total of 498 patients who could be evaluated were enrolled. Treatment intensity was based on presenting features and the level of minimal residual disease after remission-induction treatment. The duration of continuous complete remission in the 71 patients who previously would have received prophylactic cranial irradiation was compared with that of 56 historical controls who received it. RESULTS: The 5-year event-free and overall survival probabilities for all 498 patients were 85.6% (95% confidence interval [CI], 79.9 to 91.3) and 93.5% (95% CI, 89.8 to 97.2), respectively. The 5-year cumulative risk of isolated CNS relapse was 2.7% (95% CI, 1.1 to 4.3), and that of any CNS relapse (including isolated relapse and combined relapse) was 3.9% (95% CI, 1.9 to 5.9). The 71 patients had significantly longer continuous complete remission than the 56 historical controls (P=0.04). All 11 patients with isolated CNS relapse remained in second remission for 0.4 to 5.5 years. CNS leukemia (CNS-3 status) or a traumatic lumbar puncture with blast cells at diagnosis and a high level of minimal residual disease (> or = 1%) after 6 weeks of remission induction were significantly associated with poorer event-free survival. Risk factors for CNS relapse included the genetic abnormality t(1;19)(TCF3-PBX1), any CNS involvement at diagnosis, and T-cell immunophenotype. Common adverse effects included allergic reactions to asparaginase, osteonecrosis, thrombosis, and disseminated fungal infection. CONCLUSIONS: With effective risk-adjusted chemotherapy, prophylactic cranial irradiation can be safely omitted from the treatment of childhood ALL. (ClinicalTrials.gov number, NCT00137111.) PMID- 19553646 TI - Routine early angioplasty after fibrinolysis for acute myocardial infarction. AB - BACKGROUND: Patients with a myocardial infarction with ST-segment elevation who present to hospitals that do not have the capability of performing percutaneous coronary intervention (PCI) often cannot undergo timely primary PCI and therefore receive fibrinolysis. The role and optimal timing of routine PCI after fibrinolysis have not been established. METHODS: We randomly assigned 1059 high risk patients who had a myocardial infarction with ST-segment elevation and who were receiving fibrinolytic therapy at centers that did not have the capability of performing PCI to either standard treatment (including rescue PCI, if required, or delayed angiography) or a strategy of immediate transfer to another hospital and PCI within 6 hours after fibrinolysis. All patients received aspirin, tenecteplase, and heparin or enoxaparin; concomitant clopidogrel was recommended. The primary end point was the composite of death, reinfarction, recurrent ischemia, new or worsening congestive heart failure, or cardiogenic shock within 30 days. RESULTS: Cardiac catheterization was performed in 88.7% of the patients assigned to standard treatment a median of 32.5 hours after randomization and in 98.5% of the patients assigned to routine early PCI a median of 2.8 hours after randomization. At 30 days, the primary end point occurred in 11.0% of the patients who were assigned to routine early PCI and in 17.2% of the patients assigned to standard treatment (relative risk with early PCI, 0.64; 95% confidence interval, 0.47 to 0.87; P=0.004). There were no significant differences between the groups in the incidence of major bleeding. CONCLUSIONS: Among high-risk patients who had a myocardial infarction with ST-segment elevation and who were treated with fibrinolysis, transfer for PCI within 6 hours after fibrinolysis was associated with significantly fewer ischemic complications than was standard treatment. (ClinicalTrials.gov number, NCT00164190.) PMID- 19553651 TI - Case records of the Massachusetts General Hospital. Case 20-2009. A 79-year-old woman with a blistering cutaneous eruption. PMID- 19553652 TI - Routine angioplasty after fibrinolysis--how early should "early" be? PMID- 19553653 TI - Arrhythmogenic right ventricular cardiomyopathy. PMID- 19553654 TI - Ozone exposure and mortality. PMID- 19553655 TI - Growth in care provided by hospitalists. PMID- 19553656 TI - Medical evaluation before electroconvulsive therapy. PMID- 19553657 TI - Ethical and scientific implications of the globalization of clinical research. PMID- 19553658 TI - Molecular outcome prediction in diffuse large-B-cell lymphoma. PMID- 19553659 TI - Role of partitioning-defective 1/microtubule affinity-regulating kinases in the morphogenetic activity of Helicobacter pylori CagA. AB - Helicobacter pylori CagA plays a key role in gastric carcinogenesis. Upon delivery into gastric epithelial cells, CagA binds and deregulates SHP-2 phosphatase, a bona fide oncoprotein, thereby causing sustained ERK activation and impaired focal adhesions. CagA also binds and inhibits PAR1b/MARK2, one of the four members of the PAR1 family of kinases, to elicit epithelial polarity defect. In nonpolarized gastric epithelial cells, CagA induces the hummingbird phenotype, an extremely elongated cell shape characterized by a rear retraction defect. This morphological change is dependent on CagA-deregulated SHP-2 and is thus thought to reflect the oncogenic potential of CagA. In this study, we investigated the role of the PAR1 family of kinases in the hummingbird phenotype. We found that CagA binds not only PAR1b but also other PAR1 isoforms, with order of strength as follows: PAR1b > PAR1d >or= PAR1a > PAR1c. Binding of CagA with PAR1 isoforms inhibits the kinase activity. This abolishes the ability of PAR1 to destabilize microtubules and thereby promotes disassembly of focal adhesions, which contributes to the hummingbird phenotype. Consistently, PAR1 knockdown potentiates induction of the hummingbird phenotype by CagA. The morphogenetic activity of CagA was also found to be augmented through inhibition of non-muscle myosin II. Because myosin II is functionally associated with PAR1, perturbation of PAR1-regulated myosin II by CagA may underlie the defect of rear retraction in the hummingbird phenotype. Our findings reveal that CagA systemically inhibits PAR1 family kinases and indicate that malfunctioning of microtubules and myosin II by CagA-mediated PAR1 inhibition cooperates with deregulated SHP-2 in the morphogenetic activity of CagA. PMID- 19553660 TI - PinX1 is a novel microtubule-binding protein essential for accurate chromosome segregation. AB - Mitosis is an orchestration of dynamic interactions between spindle microtubules and chromosomes, which is mediated by protein structures that include the kinetochores, and other protein complexes present on chromosomes. PinX1 is a potent telomerase inhibitor in interphase; however, its function in mitosis is not well documented. Here we show that PinX1 is essential for faithful chromosome segregation. Deconvolution microscopic analyses show that PinX1 localizes to nucleoli and telomeres in interphase and relocates to the periphery of chromosomes and the outer plate of the kinetochores in mitosis. Our deletion analyses mapped the kinetochore localization domain of PinX1 to the central region and its chromosome periphery localization domain to the C terminus. Interestingly, the kinetochore localization of PinX1 is dependent on Hec1 and CENP-E. Our biochemical characterization revealed that PinX1 is a novel microtubule-binding protein. Our real time imaging analyses show that suppression of PinX1 by small interference RNA abrogates faithful chromosome segregation and results in anaphase chromatid bridges in mitosis and micronuclei in interphase, suggesting an essential role of PinX1 in chromosome stability. Taken together, the results indicate that PinX1 plays an important role in faithful chromosome segregation in mitosis. PMID- 19553661 TI - Unusual water-mediated antigenic recognition of the proinflammatory cytokine interleukin-18. AB - The unique cytokine interleukin-18 (IL-18) acts synergistically with IL-12 to regulate T-helper 1 and 2 lymphocytes and, as such, seems to underlie the pathogenesis of various autoimmune and allergic diseases. Several anti-IL-18 agents are in clinical development, including the recombinant human antibody ABT 325, which is entering trials for autoimmune diseases. Given competing cytokine/receptor and cytokine/receptor decoy interactions, understanding the structural basis for recognition is critical for effective development of anti cytokine therapies. Here we report three crystal structures: the murine antibody 125-2H Fab fragment bound to human IL-18, at 1.5 A resolution; the 125-2H Fab (2.3 A); and the ABT-325 Fab (1.5 A). These structures, along with human/mouse IL 18 chimera binding data, allow us to make three key observations relevant to the biology and antigenic recognition of IL-18 and related cytokines. First, several IL-18 residues shift dramatically (> 10 A) upon binding 125-2H, compared with unbound IL-18 (Kato, Z., Jee, J., Shikano, H., Mishima, M., Ohki, I., Ohnishi, H., Li, A., Hashimoto, K., Matsukuma, E., Omoya, K., Yamamoto, Y., Yoneda, T., Hara, T., Kondo, N., and Shirakawa, M. (2003) Nat. Struct. Biol. 10, 966-971). IL 18 thus exhibits plasticity that may be common to its interactions with other receptors. Related cytokines may exhibit similar plasticity. Second, ABT-325 and 125-2H differ significantly in combining site character and architecture, thus explaining their ability to bind IL-18 simultaneously at distinct epitopes. These data allow us to define the likely ABT-325 epitope and thereby explain the distinct neutralizing mechanisms of both antibodies. Third, given the high 125-2H potency, 10 well ordered water molecules are trapped upon complex formation in a cavity between two IL-18 loops and all six 125-2H complementarity-determining regions. Thus, counterintuitively, tight and specific antibody binding may in some cases be water-mediated. PMID- 19553663 TI - Mechanism of Epac activation: structural and functional analyses of Epac2 hinge mutants with constitutive and reduced activities. AB - Epac2 is a member of the family of exchange proteins directly activated by cAMP (Epac). Our previous studies suggest a model of Epac activation in which cAMP binding to the enzyme induces a localized "hinge" motion that reorients the regulatory lobe relative to the catalytic lobe without inducing large conformational changes within individual lobes. In this study, we identified the location of the major hinge in Epac2 by normal mode motion correlation and structural alignment analyses. Targeted mutagenesis was then performed to test the functional importance of hinge bending for Epac activation. We show that substitution of the conserved residue phenylalanine 435 with glycine (F435G) facilitates the hinge bending and leads to a constitutively active Epac2 capable of stimulating nucleotide exchange in the absence of cAMP. In contrast, substitution of the same residue with a bulkier side chain, tryptophan (F435W), impedes the hinge motion and results in a dramatic decrease in Epac2 catalytic activity. Structural parameters determined by small angle x-ray scattering further reveal that whereas the F435G mutant assumes a more extended conformation in the absence of cAMP, the F435W mutant is incapable of adopting the fully extended and active conformation in the presence of cAMP. These findings demonstrate the importance of hinge motion in Epac activation. Our study also suggests that phenylalanine at position 435 is the optimal size side chain to keep Epac closed and inactive in the absence of cAMP while still allowing the proper hinge motion for full Epac extension and activation in the presence of cAMP. PMID- 19553662 TI - Angiopoietin 2 mediates the differentiation and migration of neural progenitor cells in the subventricular zone after stroke. AB - Ischemic stroke stimulates neurogenesis in the adult rodent brain. The molecules underlying stroke-induced neurogenesis have not been fully investigated. Using real-time reverse transcription-PCR, we found that stroke substantially up regulated angiopoietin 2 (ANG2), a proangiogenic gene, expression in subventricular zone neural progenitor cells. Incubation of neural progenitor cells with recombinant human ANG2 significantly increased the number of beta-III tubulin-positive cells, a marker of immature neurons, but did not alter the number of glial fibrillary acidic protein (GFAP)-positive cells, a marker of astrocytes, suggesting that ANG2 promotes neuronal differentiation. Blockage of the ANG2 receptor, Tie2, with small interference RNA (siRNA)-Tie2 attenuated recombinant human ANG2 (rhANG2)-increased beta-III tubulin mRNA levels compared with levels in the progenitor cells transfected with control siRNA. Chromatin immunoprecipitation analysis revealed that CCAAT/enhancer-binding protein (C/EBP beta) up-regulated by rhANG2 bound to beta-III tubulin, which is consistent with published data that there are several C/EBP beta binding sites in the promoter of beta-III tubulin gene. In addition, rhANG2 enhanced migration of neural progenitor cells measured by single neurosphere assay. Blockage of Tie2 with siRNA-Tie2 and a Tie2-neutralizing antibody did not suppress ANG2-enhanced migration. However, inhibition of matrix metalloproteinases with GM6001 blocked ANG2-enhanced migration. Collectively, our data suggest that interaction of ANG2, a proangiogenic factor, with its receptor Tie2 promotes neural progenitor cell differentiation into neuronal lineage cells, whereas ANG2 regulates neural progenitor cell migration through matrix metalloproteinases, which do not require its receptor Tie2. PMID- 19553664 TI - Bile acid-induced epidermal growth factor receptor activation in quiescent rat hepatic stellate cells can trigger both proliferation and apoptosis. AB - Bile acids have been reported to induce epidermal growth factor receptor (EGFR) activation and subsequent proliferation of activated hepatic stellate cells (HSC), but the underlying mechanisms and whether quiescent HSC are also a target for bile acid-induced proliferation or apoptosis remained unclear. Therefore, primary rat HSC were cultured for up to 48 h and analyzed for their proliferative/apoptotic responses toward bile acids. Hydrophobic bile acids, i.e. taurolithocholate 3-sulfate, taurochenodeoxycholate, and glycochenodeoxycholate, but not taurocholate or tauroursodeoxycholate, induced Yes-dependent EGFR phosphorylation. Simultaneously, hydrophobic bile acids induced phosphorylation of the NADPH oxidase subunit p47(phox) and formation of reactive oxygen species (ROS). ROS production was sensitive to inhibition of acidic sphingomyelinase, protein kinase Czeta, and NADPH oxidases. All maneuvers which prevented bile acid induced ROS formation also prevented Yes and subsequent EGFR phosphorylation. Taurolithocholate 3-sulfate-induced EGFR activation was followed by extracellular signal-regulated kinase 1/2, but not c-Jun N-terminal kinase (JNK) activation, and stimulated HSC proliferation. When, however, a JNK signal was induced by coadministration of cycloheximide or hydrogen peroxide (H2O2), activated EGFR associated with CD95 and triggered EGFR-mediated CD95-tyrosine phosphorylation and subsequent formation of the death-inducing signaling complex. In conclusion, hydrophobic bile acids lead to a NADPH oxidase-driven ROS generation followed by a Yes-mediated EGFR activation in quiescent primary rat HSC. This proliferative signal shifts to an apoptotic signal when a JNK signal simultaneously comes into play. PMID- 19553665 TI - Characterization of wise protein and its molecular mechanism to interact with both Wnt and BMP signals. AB - Cross-talk of BMP and Wnt signaling pathways has been implicated in many aspects of biological events during embryogenesis and in adulthood. A secreted protein Wise and its orthologs (Sostdc1, USAG-1, and Ectodin) have been shown to modulate Wnt signaling and also inhibit BMP signals. Modulation of Wnt signaling activity by Wise is brought about by an interaction with the Wnt co-receptor LRP6, whereas BMP inhibition is by binding to BMP ligands. Here we have investigated the mode of action of Wise on Wnt and BMP signals. It was found that Wise binds LRP6 through one of three loops formed by the cystine knot. The Wise deletion construct lacking the LRP6-interacting loop domain nevertheless binds BMP4 and inhibits BMP signals. Moreover, BMP4 does not interfere with Wise-LRP6 binding, suggesting separate domains for the physical interaction. Functional assays also show that the ability of Wise to block Wnt1 activity through LRP6 is not impeded by BMP4. In contrast, the ability of Wise to inhibit BMP4 is prevented by additional LRP6, implying a preference of Wise in binding LRP6 over BMP4. In addition to the interaction of Wise with BMP4 and LRP6, the molecular characteristics of Wise, such as glycosylation and association with heparan sulfate proteoglycans on the cell surface, are suggested. This study helps to understand the multiple functions of Wise at the molecular level and suggests a possible role for Wise in balancing Wnt and BMP signals. PMID- 19553666 TI - The charged linker region is an important regulator of Hsp90 function. AB - Hsp90 is an ATP-dependent molecular chaperone which assists the maturation of a large set of target proteins. Members of the highly conserved Hsp90 family are found from bacteria to higher eukaryotes, with homologues in different organelles. The core architecture of Hsp90 is defined by the N-terminal ATP binding domain followed by the middle domain and the C-terminal dimerization domain. A long, highly charged linker between the N-terminal domain and the middle domain is a feature characteristic for Hsp90s of eukaryotic organisms. We set out to clarify the function of this linker by studying the effects of deletions in this region in vivo and in vitro. Here we show that increasing deletions in the charged linker region lead to defects ranging from mild temperature sensitivity to a lethal phenotype. The lethal deletion variants investigated in this study still exhibit ATPase activity. Deletion of the charged linker ultimately causes a loss of Hsp90 regulation by co-chaperones, as the sensitivity for Aha1-mediated ATPase acceleration declines, and binding of p23/Sba1 is lost in non-viable deletion constructs. In vivo client assays additionally demonstrated that the deletion of the linker had a pronounced effect on the ability of Hsp90 to facilitate client activation. A partial reconstruction of the linker sequence showed that the supplementation by artificial sequences can rescue the functionality of Hsp90 and restore the conformational flexibility of the protein, required for the processing of client proteins. PMID- 19553667 TI - A progesterone receptor co-activator (JDP2) mediates activity through interaction with residues in the carboxyl-terminal extension of the DNA binding domain. AB - Progesterone receptor (PR) belongs to the nuclear receptor family of ligand dependent transcription factors and mediates the major biological effects of progesterone. Transcriptional co-activators that are recruited by PR through the carboxyl-terminal ligand binding domain have been studied extensively. Much less is known about co-activators that interact with other regions of receptors. Jun dimerization protein 2 (JDP2) is a PR co-activator that enhances the transcriptional activity of the amino-terminal domain by increasing the alpha helical content and stability of the intrinsically disordered amino-terminal domain. To gain insights into the mechanism of JDP2 co-activation of PR, the structural basis of JDP2-PR interaction was analyzed using NMR. The smallest regions of each protein needed for efficient protein interaction were used for NMR and included the basic region plus leucine zipper (bZIP) domain of JDP2 and the core zinc modules of the PR DNA binding domain plus the intrinsically disordered carboxyl-terminal extension (CTE) of the DNA binding domain. Chemical shift changes in PR upon titration with JDP2 revealed that most of the residues involved in binding of JDP2 reside within the CTE. The importance of the CTE for binding JDP2 was confirmed by peptide competition and mutational analyses. Point mutations within CTE sites identified by NMR and a CTE domain swapping experiment also confirmed the functional importance of JDP2 interaction with the CTE for enhancement of PR transcriptional activity. These studies provide insights into the role and functional importance of the CTE for co-activator interactions. PMID- 19553669 TI - Hepcidin revisited, disulfide connectivity, dynamics, and structure. AB - Hepcidin is a tightly folded 25-residue peptide hormone containing four disulfide bonds, which has been shown to act as the principal regulator of iron homeostasis in vertebrates. We used multiple techniques to demonstrate a disulfide bonding pattern for hepcidin different from that previously published. All techniques confirmed the following disulfide bond connectivity: Cys(1)-Cys(8), Cys(3) Cys(6), Cys(2)-Cys(4), and Cys(5)-Cys(7). NMR studies reveal a new model for hepcidin that, at ambient temperatures, interconverts between two different conformations, which could be individually resolved by temperature variation. Using these methods, the solution structure of hepcidin was determined at 325 and 253 K in supercooled water. X-ray analysis of a co-crystal with Fab appeared to stabilize a hepcidin conformation similar to the high temperature NMR structure. PMID- 19553668 TI - Loss of NF-kappaB control and repression of Prdx6 gene transcription by reactive oxygen species-driven SMAD3-mediated transforming growth factor beta signaling. PMID- 19553670 TI - Structure-function analysis of inositol hexakisphosphate-induced autoprocessing in Clostridium difficile toxin A. AB - The action of Clostridium difficile toxins A and B depends on inactivation of host small G-proteins by glucosylation. Cellular inositol hexakisphosphate (InsP6) induces an autocatalytic cleavage of the toxins, releasing an N-terminal glucosyltransferase domain into the host cell cytosol. We have defined the cysteine protease domain (CPD) responsible for autoprocessing within toxin A (TcdA) and report the 1.6 A x-ray crystal structure of the domain bound to InsP6. InsP6 is bound in a highly basic pocket that is separated from an unusual active site by a beta-flap structure. Functional studies confirm an intramolecular mechanism of cleavage and highlight specific residues required for InsP6-induced TcdA processing. Analysis of the structural and functional data in the context of sequences from similar and diverse origins highlights a C-terminal extension and a pi-cation interaction within the beta-flap that appear to be unique among the large clostridial cytotoxins. PMID- 19553671 TI - The phox domain of sorting nexin 5 lacks phosphatidylinositol 3-phosphate (PtdIns(3)P) specificity and preferentially binds to phosphatidylinositol 4,5 bisphosphate (PtdIns(4,5)P2). AB - Subcellular retrograde transport of cargo receptors from endosomes to the trans Golgi network is critically involved in a broad range of physiological and pathological processes and highly regulated by a genetically conserved heteropentameric complex, termed retromer. Among the retromer components identified in mammals, sorting nexin 5 and 1 (SNX5; SNX1) have recently been found to interact, possibly controlling the membrane binding specificity of the complex. To elucidate how the unique sequence features of the SNX5 phox domain (SNX5-PX) influence retrograde transport, we have determined the SNX5-PX structure by NMR and x-ray crystallography at 1.5 A resolution. Although the core fold of SNX5-PX resembles that of other known PX domains, we found novel structural features exclusive to SNX5-PX. It is most noteworthy that in SNX5-PX, a long helical hairpin is added to the core formed by a new alpha2'-helix and a much longer alpha3-helix. This results in a significantly altered overall shape of the protein. In addition, the unique double PXXP motif is tightly packed against the rest of the protein, rendering this part of the structure compact, occluding parts of the putative phosphatidylinositol (PtdIns) binding pocket. The PtdIns binding and specificity of SNX5-PX was evaluated by NMR titrations with eight different PtdIns and revealed that SNX5-PX preferentially and specifically binds to phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)). The distinct structural and PtdIns binding characteristics of SNX5-PX impart specific properties on SNX5, influencing retromer-mediated regulation of retrograde trafficking of transmembrane cargo receptors. PMID- 19553672 TI - Complex glycan catabolism by the human gut microbiota: the Bacteroidetes Sus-like paradigm. AB - Trillions of microbes inhabit the distal gut of adult humans. They have evolved to compete efficiently for nutrients, including a wide array of chemically diverse, complex glycans present in our diets, secreted by our intestinal mucosa, and displayed on the surfaces of other gut microbes. Here, we review how members of the Bacteroidetes, one of two dominant gut-associated bacterial phyla, process complex glycans using a series of similarly patterned, cell envelope-associated multiprotein systems. These systems provide insights into how gut, as well as terrestrial and aquatic, Bacteroidetes survive in highly competitive ecosystems. PMID- 19553673 TI - Sterol-mediated regulation of human lipin 1 gene expression in hepatoblastoma cells. AB - Lipin 1 plays a crucial role in lipid metabolism in adipose tissue, skeletal muscle, and liver. Its physiological role involves two cellular functions: regulation of phosphatidate phosphatase activity and regulation of fatty acid oxidation. In this study, we have demonstrated that lipin 1 gene (LPIN1) expression is regulated by cellular sterols, which are key regulators of lipid metabolism. We have also characterized the sterol-response element and nuclear factor Y-binding sites in the human LPIN1 promoter. Using a luciferase assay, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay, we demonstrated that these elements are responsible for the transcription of LPIN1 gene, mediated by SREBP-1 (sterol regulatory element-binding protein 1) and nuclear factor Y. Furthermore, we investigated whether lipin 1 is involved in lipogenesis by transfection of LPIN1 small interfering RNA. We infer that sterol mediated regulation of lipin 1 gene transcription modulates triglyceride accumulation. This modulation involves changes in the activity of phosphatidate phosphatase. PMID- 19553674 TI - Carnitine insufficiency caused by aging and overnutrition compromises mitochondrial performance and metabolic control. AB - In addition to its essential role in permitting mitochondrial import and oxidation of long chain fatty acids, carnitine also functions as an acyl group acceptor that facilitates mitochondrial export of excess carbons in the form of acylcarnitines. Recent evidence suggests carnitine requirements increase under conditions of sustained metabolic stress. Accordingly, we hypothesized that carnitine insufficiency might contribute to mitochondrial dysfunction and obesity related impairments in glucose tolerance. Consistent with this prediction whole body carnitine diminution was identified as a common feature of insulin-resistant states such as advanced age, genetic diabetes, and diet-induced obesity. In rodents fed a lifelong (12 month) high fat diet, compromised carnitine status corresponded with increased skeletal muscle accumulation of acylcarnitine esters and diminished hepatic expression of carnitine biosynthetic genes. Diminished carnitine reserves in muscle of obese rats was accompanied by marked perturbations in mitochondrial fuel metabolism, including low rates of complete fatty acid oxidation, elevated incomplete beta-oxidation, and impaired substrate switching from fatty acid to pyruvate. These mitochondrial abnormalities were reversed by 8 weeks of oral carnitine supplementation, in concert with increased tissue efflux and urinary excretion of acetylcarnitine and improvement of whole body glucose tolerance. Acetylcarnitine is produced by the mitochondrial matrix enzyme, carnitine acetyltransferase (CrAT). A role for this enzyme in combating glucose intolerance was further supported by the finding that CrAT overexpression in primary human skeletal myocytes increased glucose uptake and attenuated lipid induced suppression of glucose oxidation. These results implicate carnitine insufficiency and reduced CrAT activity as reversible components of the metabolic syndrome. PMID- 19553675 TI - Regulation of Na,K-ATPase subunit abundance by translational repression. AB - The Na,K-ATPase is an alphabeta heterodimer responsible for maintaining fluid and electrolyte homeostasis in mammalian cells. We engineered Madin-Darby canine kidney cell lines expressing alpha(1)FLAG, beta(1)FLAG, or beta(2)MYC subunits via a tetracycline-regulated promoter and a line expressing both stable beta(1)MYC and tetracycline-regulated beta(1)FLAG to examine regulatory mechanisms of sodium pump subunit expression. When overexpression of exogenous beta(1)FLAG increased total beta subunit levels by >200% without changes in alpha subunit abundance, endogenous beta(1) subunit (beta(1)E) abundance decreased. beta(1)E down-regulation did not occur during beta(2)MYC overexpression, indicating isoform specificity of the repression mechanism. Measurements of RNA stability and content indicated that decreased beta subunit expression was not accompanied by any change in mRNA levels. In addition, the degradation rate of beta subunits was not altered by beta(1)FLAG overexpression. Cells stably expressing beta(1)MYC, when induced to express beta(1)FLAG subunits, showed reduced beta(1)MYC and beta(1)E subunit abundance, indicating that these effects occur via the coding sequences of the down-regulated polypeptides. In a similar way, Madin-Darby canine kidney cells overexpressing exogenous alpha(1)FLAG subunits exhibited a reduction of endogenous alpha(1) subunits (alpha(1)E) with no change in alpha mRNA levels or beta subunits. The reduction in alpha(1)E compensated for alpha(1)FLAG subunit expression, resulting in unchanged total alpha subunit abundance. Thus, regulation of alpha subunit expression maintained its native level, whereas beta subunit was not as tightly regulated and its abundance could increase substantially over native levels. These effects also occurred in human embryonic kidney cells. These data are the first indication that cellular sodium pump subunit abundance is modulated by translational repression. This mechanism represents a novel, potentially important mechanism for regulation of Na,K-ATPase expression. PMID- 19553676 TI - Bone morphogenetic protein 15 (BMP15) acts as a BMP and Wnt inhibitor during early embryogenesis. AB - Bone morphogenetic protein 15 (BMP15) belongs to an unusual subgroup of the transforming growth factor beta (TGFbeta) superfamily of signaling ligands as it lacks a key cysteine residue in the mature region required for proper intermolecular dimerization. Naturally occurring BMP15 mutation leads to early ovarian failure in humans, and BMP15 has been shown to activate the Smad1/5/8 pathway in that context. Despite its important role in germ cell specification, the embryological function of BMP15 remains unknown. Surprisingly, we find that during early Xenopus embryogenesis BMP15 acts solely as an inhibitor of the Smad1/5/8 pathway and the Wnt pathway. BMP15 gain-of-function leads to embryos with secondary ectopic heads and to direct neural induction in intact explants. BMP15 inhibits BMP4-mediated epidermal induction in dissociated explants. BMP15 strongly inhibits BRE response induced by BMP4 and blocks phosphorylation and activation of Smad1/5/8 MH2-domain. Mechanistically, BMP15 protein specifically interacts with BMP4 protein, suggesting inhibition upstream of receptor binding. Loss-of-function experiments using morpholinos or a naturally occurring human BMP15 dominant-negative mutant (BMP15-Y235C) leads to embryos lacking head. BMP15 Y235C also eliminates the inhibitory activity of BMP15 on BRE (BMP-responsive element). Finally, we show that BMP15 inhibits the canonical branch of the Wnt pathway, upstream of beta-catenin. We, thus, demonstrate that BMP15 is necessary and sufficient for the specification of dorso-anterior structures and highlight novel mechanisms of BMP15 function that strongly suggest a reinterpretation of its function in ovaries specially for ovarian failure. PMID- 19553677 TI - MRG15 is a novel PALB2-interacting factor involved in homologous recombination. AB - PALB2 is an integral component of the BRCA complex important for recombinational DNA repair. However, exactly how this activity is regulated in vivo remains unexplored. Here we provide evidence to show that MRG15 is a novel PALB2 associated protein that ensures regulated recombination events. We found that the direct interaction between MRG15 and PALB2 is mediated by an evolutionarily conserved region on PALB2. Intriguingly, although damage-induced RAD51 foci formation and mitomycin C sensitivity appeared normal in MRG15-binding defective PALB2 mutants, these cells exhibited a significant increase in gene conversion rates. Consistently, we found that abrogation of the PALB2-MRG15 interaction resulted in elevated sister chromatid exchange frequencies. Our results suggest that loss of the PALB2-MRG15 interaction relieved the cells with the suppression of sister chromatid exchange and therefore led to a hyper-recombination phenotype in the gene conversion assay. Together, our study indicated that although PALB2 is required for proficient homologous recombination, it could also govern the choice of templates used in homologous recombination repair. PMID- 19553678 TI - The activities of the Yersinia protein kinase A (YpkA) and outer protein J (YopJ) virulence factors converge on an eIF2alpha kinase. AB - The Yersinia protein kinase A (YpkA) and outer protein J (YopJ) are co-expressed from a single transcript and are injected directly into eukaryotic cells by the plague bacterium Yersinia pestis. When overexpressed in vertebrate or yeast cells, YpkA disrupts the actin-based cytoskeletal system by an unknown mechanism, whereas YopJ obstructs inductive chemokine expression by inhibiting MAPK and NF kappaB signaling. Previously, we showed that the fission yeast Schizosaccharomyces pombe was sensitive to the kinase activity of YpkA. Here, we screened yeast for cellular processes important for YpkA activity and found that the eIF2alpha kinases mollify the toxicity imparted by the kinase activity of YpkA. Specifically, strains lacking the eIF2alpha kinase Hri2 were particularly sensitive to YpkA. Unexpectedly, the activity of YopJ, which conferred a phenotype consistent with its inhibitory effect on MAPK signaling, was also found to be dependent on Hri2. When expressed in S. pombe, YopJ sensitized cells to osmotic and oxidative stresses through a Hri2-dependent mechanism. However, when co-expressed with YpkA, YopJ protected cells from YpkA-mediated toxicity, and this protection was entirely dependent on Hri2. In contrast, YopJ did not confer protection against the toxic effects of the Yersinia virulence factor YopE. These findings are the first to functionally link YpkA and YopJ and suggest that eIF2alpha kinases, which are critically important in antiviral defenses and protection against environmental stresses, also play a role in bacterial virulence. PMID- 19553679 TI - Identification of caspase-mediated decay of interferon regulatory factor-3, exploited by a Kaposi sarcoma-associated herpesvirus immunoregulatory protein. AB - Upon virus infection, the cell mounts an innate type I interferon (IFN) response to limit the spread. This response is orchestrated by the constitutively expressed IFN regulatory factor (IRF)-3 protein, which becomes post translationally activated. Although the activation events are understood in detail, the negative regulation of this innate response is less well understood. Many viruses, including Kaposi sarcoma-associated herpesvirus (KSHV), have evolved defense strategies against this IFN response. Thus, KSHV encodes a viral IRF (vIRF)-2 protein, sharing homology with cellular IRFs and is a known inhibitor of the innate IFN response. Here, we show that vIRF-2 mediates IRF-3 inactivation by a mechanism involving caspase-3, although vIRF-2 itself is not pro-apoptotic. Importantly, we also show that caspase-3 participates in normal IRF-3 turnover in the absence of vIRF-2, during the antiviral response induced by poly(I:C) transfection. These data provide unprecedented insight into negative regulation of IRF-3 following activation of the type I IFN antiviral response and the mechanism by which KSHV vIRF-2 inhibits this innate response. PMID- 19553680 TI - Oxidative stress decreases phosphatidylinositol 4,5-bisphosphate levels by deactivating phosphatidylinositol- 4-phosphate 5-kinase beta in a Syk-dependent manner. AB - Phosphatidylinositol 4,5-bisphosphate (PIP(2)) has many essential functions and its homeostasis is highly regulated. We previously found that hypertonic stress increases PIP(2) by selectively activating the beta isoform of the type I phosphatidylinositol phosphate 5-kinase (PIP5Kbeta) through Ser/Thr dephosphorylation and promoting its translocation to the plasma membrane. Here we report that hydrogen peroxide (H(2)O(2)) also induces PIP5Kbeta Ser/Thr dephosphorylation, but it has the opposite effect on PIP(2) homeostasis, PIP5Kbeta function, and the actin cytoskeleton. Brief H(2)O(2) treatments decrease cellular PIP(2) in a PIP5Kbeta-dependent manner. PIP5Kbeta is tyrosine phosphorylated, dissociates from the plasma membrane, and has decreased lipid kinase activity. In contrast, the other two PIP5K isoforms are not inhibited by H(2)O(2). We identified spleen tyrosine kinase (Syk), which is activated by oxidants, as a candidate PIP5Kbeta kinase in this pathway, and mapped the oxidant sensitive tyrosine phosphorylation site to residue 105. The PIP5KbetaY105E phosphomimetic is catalytically inactive and cytosolic, whereas the Y105F non phosphorylatable mutant has higher intrinsic lipid kinase activity and is much more membrane associated than wild type PIP5Kbeta. These results suggest that during oxidative stress, as modeled by H(2)O(2) treatment, Syk-dependent tyrosine phosphorylation of PIP5Kbeta is the dominant post-translational modification that is responsible for the decrease in cellular PIP(2). PMID- 19553682 TI - Probing HIV-1 membrane liquid order by Laurdan staining reveals producer cell dependent differences. AB - Viruses acquire their envelope by budding from a host cell membrane, but viral lipid composition may differ from that of the budding membrane. We have previously reported that the HIV-1 membrane is highly enriched in cholesterol, sphingolipids, and other raft lipids, suggesting that the virus may bud from pre existing or virus-induced lipid rafts. Here, we employed the environmentally sensitive fluorescent dye Laurdan to study the membrane lateral structure of HIV 1 derived from different cell lines. Differences in viral membrane order detected by Laurdan staining were shown by mass spectrometry to be due to differences in lipid composition. Isogenic viruses from two different cell lines were both strongly enriched in raft lipids and displayed a liquid-ordered membrane, but these effects were significantly more pronounced for HIV-1 from the T-cell line MT-4 compared with virus from 293T cells. Host-dependent differences in the lipidomes predominantly affected the ratio of sphingomyelins (including dihydrosphingomyelin) to phosphatidylcholine, whereas cholesterol contents were similar. Accordingly, treatment of infectious HIV-1 with the sphingomyelin binding toxins Equinatoxin-II or lysenin showed differential inhibition of infectivity. Liposomes consisting of lipids that had been extracted from viral particles exhibited slightly less liquid order than the respective viral membranes, which is likely to be due to absence of membrane proteins and to loss of lipid asymmetry. Synthetic liposomes consisting of a quaternary lipid mixture emulating the viral lipids showed a liquid order similar to liposomes derived from virion lipids. Thus, Laurdan staining represents a rapid and quantitative method to probe viral membrane liquid order and may prove useful in the search for lipid active drugs. PMID- 19553681 TI - ADP-dependent 6-phosphofructokinase from Pyrococcus horikoshii OT3: structure determination and biochemical characterization of PH1645. AB - Some hyperthermophilic archaea use a modified glycolytic pathway that employs an ADP-dependent glucokinase (ADP-GK) and an ADP-dependent phosphofructokinase (ADP PFK) or, in the case of Methanococcus jannaschii, a bifunctional ADP-dependent glucophosphofructokinase (ADP-GK/PFK). The crystal structures of three ADP-GKs have been determined. However, there is no structural information available for ADP-PFKs or the ADP-GK/PFK. Here, we present the first crystal structure of an ADP-PFK from Pyrococcus horikoshii OT3 (PhPFK) in both apo- and AMP-bound forms determined to 2.0-A and 1.9-A resolution, respectively, along with biochemical characterization of the enzyme. The overall structure of PhPFK maintains a similar large and small alpha/beta domain structure seen in the ADP-GK structures. A large conformational change accompanies binding of phosphoryl donor, acceptor, or both, in all members of the ribokinase superfamily characterized thus far, which is believed to be critical to enzyme function. Surprisingly, no such conformational change was observed in the AMP-bound PhPFK structure compared with the apo structure. Through comprehensive site-directed mutagenesis of the substrate binding pocket we identified residues that were critical for both substrate recognition and the phosphotransfer reaction. The catalytic residues and many of the substrate binding residues are conserved between PhPFK and ADP-GKs; however, four key residues differ in the sugar-binding pocket, which we have shown determine the sugar-binding specificity. Using these results we were able to engineer a mutant PhPFK that mimics the ADP-GK/PFK and is able to phosphorylate both fructose 6-phosphate and glucose. PMID- 19553683 TI - Myosin II tailpiece determines its paracrystal structure, filament assembly properties, and cellular localization. AB - Non muscle myosin II (NMII) is a major motor protein present in all cell types. The three known vertebrate NMII isoforms share high sequence homology but play different cellular roles. The main difference in sequence resides in the C terminal non-helical tailpiece (tailpiece). In this study we demonstrate that the tailpiece is crucial for proper filament size, overcoming the intrinsic properties of the coiled-coil rod. Furthermore, we show that the tailpiece by itself determines the NMII filament structure in an isoform-specific manner, thus providing a possible mechanism by which each NMII isoform carries out its unique cellular functions. We further show that the tailpiece determines the cellular localization of NMII-A and NMII-B and is important for NMII-C role in focal adhesion complexes. We mapped NMII-C sites phosphorylated by protein kinase C and casein kinase II and showed that these phosphorylations affect its solubility properties and cellular localization. Thus phosphorylation fine-tunes the tailpiece effects on the coiled-coil rod, enabling dynamic regulation of NMII-C assembly. We thus show that the small tailpiece of NMII is a distinct domain playing a role in isoform-specific filament assembly and cellular functions. PMID- 19553684 TI - SIRT1 controls the transcription of the peroxisome proliferator-activated receptor-gamma Co-activator-1alpha (PGC-1alpha) gene in skeletal muscle through the PGC-1alpha autoregulatory loop and interaction with MyoD. AB - Peroxisome proliferator activated receptor-gamma co-activator-1alpha (PGC-1alpha) is a transcriptional co-activator that coordinately regulates the expression of distinct sets of metabolism-related genes in different tissues. Here we show that PGC-1alpha expression is reduced in skeletal muscles from mice lacking the sirtuin family deacetylase SIRT1. Conversely, SIRT1 activation or overexpression in differentiated C2C12 myotubes increased PGC-1alpha mRNA expression. The transcription-promoting effects of SIRT1 occurred through stimulation of PGC 1alpha promoter activity and were enhanced by co-transfection of myogenic factors, such as myocyte enhancer factor 2 (MEF2) and, especially, myogenic determining factor (MyoD). SIRT1 bound to the proximal promoter region of the PGC 1alpha gene, an interaction potentiated by MEF2C or MyoD, which also interact with this region. In the presence of MyoD, SIRT1 promoted a positive autoregulatory PGC-1alpha expression loop, such that overexpression of PGC-1alpha increased PGC-1alpha promoter activity in the presence of co-expressed MyoD and SIRT1. Chromatin immunoprecipitation showed that SIRT1 interacts with PGC-1alpha promoter and increases PGC-1alpha recruitment to its own promoter region. Immunoprecipitation assays further showed that SIRT1-PGC-1alpha interactions are enhanced by MyoD. Collectively, these data indicate that SIRT1 controls PGC 1alpha gene expression in skeletal muscle and that MyoD is a key mediator of this action. The involvement of MyoD in SIRT1-dependent PGC-1alpha expression may help to explain the ability of SIRT1 to drive muscle-specific gene expression and metabolism. Autoregulatory control of PGC-1alpha gene transcription seems to be a pivotal mechanism for conferring a transcription-activating response to SIRT1 in skeletal muscle. PMID- 19553686 TI - Keeping it simple: what mouse models of Wolf-Hirschhorn syndrome can tell us about large chromosomal deletions. PMID- 19553687 TI - How to phenotype a mouse. PMID- 19553689 TI - Bringing together top young investigators at the frontiers of science. PMID- 19553685 TI - Inactivation of mammalian target of rapamycin increases STAT1 nuclear content and transcriptional activity in alpha4- and protein phosphatase 2A-dependent fashion. AB - Target of rapamycin (TOR) is a highly conserved serine/threonine kinase that controls cell growth, primarily via regulation of protein synthesis. In Saccharomyces cerevisiae, TOR can also suppress the transcription of stress response genes by a mechanism involving Tap42, a serine/threonine phosphatase subunit, and the transcription factor Msn2. A physical association between mammalian TOR (mTOR) and the transcription factor signal transducer and activator of transcription-1 (STAT1) was recently identified in human cells, suggesting a similar role for mTOR in the transcription of interferon-gamma-stimulated genes. In the current study, we identified a macromolecular protein complex composed of mTOR, STAT1, the Tap42 homologue alpha4, and the protein phosphatase 2A catalytic subunit (PP2Ac). Inactivation of mTOR enhanced its association with STAT1 and increased STAT1 nuclear content in PP2Ac-dependent fashion. Depletion of alpha4, PP2A, or mTOR enhanced the induction of early (i.e. IRF-1) and late (i.e. caspase 1, hiNOS, and Fas) STAT1-dependent genes. The regulation of IRF-1 or caspase-1 by mTOR was independent of other known mTOR effectors p70 S6 kinase and Akt. These results describe a new role for mTOR and alpha4/PP2A in the control of STAT1 nuclear content, and the expression of interferon-gamma-sensitive genes involved in immunity and apoptosis. PMID- 19553691 TI - Anti-angiogenic drugs to treat human disease: an interview with Napoleone Ferrara by Kristin H. Kain. PMID- 19553692 TI - Researchers focus on inflammation at the Istituto Clinico Humanitas. PMID- 19553693 TI - Developing an animal model for infantile spasms: pathogenesis, problems and progress. AB - Infantile spasms (IS), the most common of the early epileptic encephalopathies, afflicts thousands of children each year and results in significant disability. Also known as West syndrome, IS is characterized by intractable stereotyped seizures, poor developmental outcome and a characteristic electroencephalogram (EEG) pattern. IS often progresses into another epileptic encephalopathy known as Lennox-Gastaut syndrome, and continues with the patient being burdened by lifelong epilepsy and varying degrees of mental retardation. Little is known about the biological basis of IS. As the etiologies of IS are diverse, the multiple causes must converge into a final common pathway that results in this specific epilepsy phenotype. Finding a model or models to test this final pathway is necessary both to understand why the greatest susceptibility to seizure development occurs during infancy and early childhood, and what underlies the decreased cognitive potential associated with IS. Furthermore, appropriate models would permit better testing of potential therapies directed specifically at IS. This review will describe the clinical features and etiologies of IS; the ideal features that IS models should contain; and the IS models that exist currently. Finally, we will discuss the limitations of these models and the potential avenues for future research on IS. PMID- 19553694 TI - Mitochondrial dysfunction and Parkinson's disease genes: insights from Drosophila. AB - Parkinson's disease (PD), one of the most common neurodegenerative disorders worldwide, currently lacks a cure. Although most PD cases occur sporadically, studies from rare genetic mutations give significant insights into addressing the pathological mechanism of not only familial PD, but also sporadic PD. Recent PD research focuses on generating genetic mutant animal models that recapitulate the features of human PD patients. Significant advances in PD research have resulted from studying Drosophila mutants of several identified PD-associated genes because they show strikingly visible phenotypes. In particular, previous studies with the Drosophila mutants parkin and PINK1, which are two common causative genes among PD familial forms, have suggested strongly that mitochondrial dysfunction is the prominent cause for the PD pathogenesis and that these two PD genes are in a common pathway, with Parkin downstream of PINK1. Recent genetic studies have revealed that the PINK1-Parkin pathway is involved in regulating the mitochondrial remodeling process. In addition, PINK1 was recently found to regulate the localization of Parkin through direct phosphorylation. Here, we briefly review these new and exciting findings in Drosophila PD models and discuss how using these models can further advance PD studies. PMID- 19553695 TI - Pluripotent stem cells and other technologies will eventually open the door for straightforward gene targeting in the rat. AB - Although the rat is a preferred model in many fields of biomedical sciences, the inability to generate germline competent embryonic stem (ES) cells was a major drawback for research activities that aimed to elucidate gene functions. Several alternative strategies like N-ethyl-N-nitrosourea (ENU) or transposon-mediated mutagenesis were developed successfully for this species. Countless experiments in many laboratories around the world were undertaken to overcome this problem. Eventually, the successful establishment of rat ES cells and rat-induced pluripotent stem (iPS) cells was reported, 27 years after the first reported generation of mouse ES cells. Furthermore, the application of zinc-finger nucleases (ZFNs) to early-stage rat embryos demonstrated effectively that another way existed for generating knockout rats. ZFNs require only the standard techniques that are used to produce transgenic animals and are expected to comprise a major tool for the gene-targeted generation of knockout animals. These newly developed tools, in conjunction with the solid basis of the rat in the area of physiological and behavioral experiments, will not only close the gap between the rat and the mouse as the mammalian genetic model of choice, but also boost the significance of the rat as a model animal in research laboratories around the globe. PMID- 19553697 TI - Human term placenta a new abundant source of hematopoietic cells--a potent alternative for cord blood and bone marrow. PMID- 19553698 TI - Convergence of the 5-LOX and COX-2 pathways: heme-catalyzed cleavage of the 5S HETE-derived di-endoperoxide into aldehyde fragments. AB - Oxygenation of the 5-lipoxygenase product 5S-hydroxyeicosatetraenoic acid by cyclooxygenase-2 yields a bicyclic di-endoperoxide. The di-endoperoxide contains two peroxides spanning from carbons 9 to 11 and 8 to 12, and two hydroxyls at carbons 5 and 15 of arachidonic acid (Schneider C., et al. 2006. Convergent oxygenation of arachidonic acid by 5-lipoxygenase and cyclooxygenase-2. J. Am. Chem. Soc. 128: 720). Here, we report that treatment of the di-endoperoxide with hematin or ferrous chloride results in cleavage of both peroxide O-O bonds and of the bonds between the carbons that carry the peroxide groups, producing the aldehydes 4-hydroxy-2E-nonenal (4-HNE), 8-oxo-5S-hydroxy-6E-octenoic acid, and malondialdehyde (MDA). The hematin- and ferrous iron-catalyzed transformation of the di-endoperoxide proceeded with a similar yield of products as the cleavage of the prostaglandin endoperoxide PGH(2) to 12S-hydroxy-5Z,8E,10E-heptadecatrienoic acid and MDA. Chiral phase HPLC analysis of the 4-HNE cleavage product showed greater than 98% 4S and thus established the S configuration of the 15-carbon of the di-endoperoxide that had not previously been assigned. This transformation of the 5-lipoxygenase/cyclooxygenase-2 derived di-endoperoxide invokes the possibility of a novel pathway to formation of the classic lipid peroxidation products 4-HNE and MDA. PMID- 19553699 TI - Crystal structure of the LG1-3 region of the laminin alpha2 chain. AB - Laminins are large heterotrimeric glycoproteins with many essential functions in basement membrane assembly and function. Cell adhesion to laminins is mediated by a tandem of five laminin G-like (LG) domains at the C terminus of the alpha chain. Integrin binding requires an intact LG1-3 region, as well as contributions from the coiled coil formed by the alpha, beta, and gamma chains. We have determined the crystal structure at 2.8-A resolution of the LG1-3 region of the laminin alpha2 chain (alpha 2LG1-3). The three LG domains adopt typical beta sandwich folds, with canonical calcium binding sites in LG1 and LG2. LG2 and LG3 interact through a substantial interface, but LG1 is completely dissociated from the LG2-3 pair. We suggest that the missing gamma chain tail may be required to stabilize the interaction between LG1 and LG2-3 in the biologically active conformation. A global analysis of N-linked glycosylation sites shows that the beta-sandwich faces of LG1 are free of carbohydrate modifications in all five laminin alpha chains, suggesting that these surfaces may harbor the integrin binding site. The alpha 2LG1-3 structure provides the first atomic view of the integrin binding region of laminins. PMID- 19553700 TI - Angioinhibitory action of NK4 involves impaired extracellular assembly of fibronectin mediated by perlecan-NK4 association. AB - NK4, a fragment of hepatocyte growth factor (HGF), exerts bifunctional action as a competitive antagonist against HGF and its receptor c-Met and an angiogenesis inhibitor. Here we studied the anti-angiogenic mechanism of NK4. In cultured human endothelial cells, NK4 inhibited DNA synthesis induced not only by HGF but also by either basic fibroblast growth factor or vascular endothelial growth factor. Even if c-Met expression was diminished by small interference RNA, NK4 inhibited basic fibroblast growth factor-induced DNA synthesis, indicating that anti-angiogenic action of NK4 is c-Met-independent. Affinity purification with NK4-immobilized beads revealed that NK4 binds to perlecan. Consistent with this, NK4 colocalized with perlecan in endothelial cells. Perlecan is a multidomain heparan sulfate proteoglycan that interacts with basement membrane components such as fibronectin. NK4 inhibited extracellular assembly of fibronectin, by which fibronectin-dependent endothelial cell spreading was inhibited by NK4. Knockdown of perlecan expression by small interference RNA significantly abrogated the inhibitory effect of NK4 on fibronectin assembly and cell spreading. In NK4-treated endothelial cells, tyrosine phosphorylation of focal adhesion kinase and Rac activation were reduced, whereas overexpression of activated Rac recovered the DNA synthesis in NK4-treated endothelial cells. These results indicate that the association between NK4 and perlecan impairs fibronectin assembly, thereby inhibiting anchorage-dependent signaling. The identified mechanism for angiostatic action provides further proof of significance for NK4 in the treatment of cancer and potentially for vascular regulation as well. PMID- 19553696 TI - Cardiac repair and regeneration: the Rubik's cube of cell therapy for heart disease. AB - Acute ischemic injury and chronic cardiomyopathies damage healthy heart tissue. Dead cells are gradually replaced by a fibrotic scar, which disrupts the normal electromechanical continuum of the ventricular muscle and compromises its pumping capacity. Recent studies in animal models of ischemic cardiomyopathy suggest that transplantation of various stem cell preparations can improve heart recovery after injury. The first clinical trials in patients produced some encouraging results, showing modest benefits. Most of the positive effects are probably because of a favorable paracrine influence of stem cells on the disease microenvironment. Stem cell therapy attenuates inflammation, reduces apoptosis of surrounding cells, induces angiogenesis, and lessens the extent of fibrosis. However, little new heart tissue is formed. The current challenge is to find ways to improve the engraftment, long-term survival and appropriate differentiation of transplanted stem cells within the cardiovascular tissue. Hence, there has been a surge of interest in pluripotent stem cells with robust cardiogenic potential, as well as in the inherent repair and regenerative mechanisms of the heart. Recent discoveries on the biology of adult stem cells could have relevance for cardiac regeneration. Here, we discuss current developments in the field of cardiac repair and regeneration, and present our ideas about the future of stem cell therapy. PMID- 19553702 TI - Validity of perinatal pharmacoepidemiologic studies using data from the RAMQ administrative database. AB - BACKGROUND: The RAMQ prescription claims database (RAMQ-Rx) is increasingly being used in perinatal pharmacoepidemiologic studies; but, there are reasons to believe that results generated with the RAMQ-Rx might not be generalizable to all patient populations. OBJECTIVES: Compare characteristics between pregnant women insured by the RAMQ-Rx and those insured by a private drug insurance plan. METHODS: A prospective study performed within the population of pregnant women receiving prenatal care at different obstetrics and gynecology clinics affiliated to the University of Montreal, Canada, was conducted from October 2004 to March 2006. Women were eligible if they were > or =18 years of age, < or =16 weeks of gestation at the time of their first prenatal visit, and able to read and understand French or English. Eligible women were asked to fill out a self administered questionnaire. RESULTS: Three hundred and sixty-three women met inclusion criteria, of which 99 (27%) had RAMQ-Rx coverage, and 264 (73%) had a private drug insurance coverage. Compared to those who were covered by private drug insurance plans, those insured by the RAMQ-Rx were younger (30.7yrs vs. 32.1yrs; P=0.03), more likely to be immigrant (60% vs. 24%; P<0.01), and have a household income below poverty level (39% vs. 2%; P<0.01). They were also less likely to be Caucasian (69% vs. 86%; P<0.01), employed (51% vs. 87%; P<.01), and have a post-secondary education (76% vs. 95%; P<0.01). No differences were observed on smoking status and alcohol use during pregnancy. CONCLUSIONS: There are substantial differences between pregnant women insured by the RAMQ-Rx and those insured by private drug insurance plans. However, these differences will most likely limit generalizability, but not internal validity, of studies using data from the RAMQ-Rx database. PMID- 19553703 TI - Ethylglucuronide and ethylsulfate in meconium to assess gestational ethanol exposure: preliminary results in two Mediterranean cohorts. AB - BACKGROUND: In recent years, fatty acid ethyl esters (FAEEs) in meconium emerged as reliable, direct biological markers for establishing gestational ethanol exposure. Among the minor nonoxidative products of ethanol metabolism, there are ethyl glucuronide (EtG) and ethyl sulfate (EtS). OBJECTIVES: The aim of the study was to analyse meconium specimens from two different Mediterranean cohorts to check for the presence of EtG and EtS, and to investigate the eventual correlation between meconium FAEEs and these two metabolites and their possible application as direct biomarkers of gestational ethanol exposure. METHODS: FAEEs, EtG and EtS were quantified by liquid chromatography tandem mass spectrometry in meconium samples obtained from the Neonatal Intensive Care Unit of Arcispedale Santa Maria Nuova, Reggio Emilia, Italy (N= 96) and from the Pediatric Service of the Hospital del Mar in Barcelona, Spain (N=81). RESULTS: EtG was present in more than 80% meconium samples while EtS only in 50% specimens Although the samples from Spain and Italy originated from similar socio-demographic cohort, EtG values in the Barcelona samples (median value: 101.5 ng/g) were statistically higher than those from Reggio Emilia ones (median value: 15.6 ng/g). In the Barcelona cohort, EtG values could differentiate between samples with FAEEs below and those equal or above 2 nmol/g - the cut-off used to differentiate heavy maternal ethanol consumption during pregnancy from occasional or no use. CONCLUSION: For the first time the presence of EtG and EtS in meconium has been proven, with EtG concentration likely to discriminate heavy maternal ethanol consumption during pregnancy disclosed by FAEEs concentration in this matrix. Further investigations are needed to verify the use of these two ethanol metabolites as alternative biomarkers of chronic in utero exposure to ethanol. PMID- 19553701 TI - The crystal structure of an algal prolyl 4-hydroxylase complexed with a proline rich peptide reveals a novel buried tripeptide binding motif. AB - Plant and algal prolyl 4-hydroxylases (P4Hs) are key enzymes in the synthesis of cell wall components. These monomeric enzymes belong to the 2-oxoglutarate dependent superfamily of enzymes characterized by a conserved jelly-roll framework. This algal P4H has high sequence similarity to the catalytic domain of the vertebrate, tetrameric collagen P4Hs, whereas there are distinct sequence differences with the oxygen-sensing hypoxia-inducible factor P4H subfamily of enzymes. We present here a 1.98-A crystal structure of the algal Chlamydomonas reinhardtii P4H-1 complexed with Zn(2+) and a proline-rich (Ser-Pro)(5) substrate. This ternary complex captures the competent mode of binding of the peptide substrate, being bound in a left-handed (poly)l-proline type II conformation in a tunnel shaped by two loops. These two loops are mostly disordered in the absence of the substrate. The importance of these loops for the function is confirmed by extensive mutagenesis, followed up by enzyme kinetic characterizations. These loops cover the central Ser-Pro-Ser tripeptide of the substrate such that the hydroxylation occurs in a highly buried space. This novel mode of binding does not depend on stacking interactions of the proline side chains with aromatic residues. Major conformational changes of the two peptide binding loops are predicted to be a key feature of the catalytic cycle. These conformational changes are probably triggered by the conformational switch of Tyr(140), as induced by the hydroxylation of the proline residue. The importance of these findings for understanding the specific binding and hydroxylation of (X Pro-Gly)(n) sequences by collagen P4Hs is also discussed. PMID- 19553704 TI - [Case of the month. A boy with mysterious rash]. PMID- 19553705 TI - [Nutritional support in the critically ill]. PMID- 19553706 TI - [Pharmaceutical expenses during economic crisis--opportunity for reform]. PMID- 19553707 TI - [Energy expenditure and nutritional support in intensive care patients]. AB - STUDY OBJECTIVES: Nutritional support of ICU patients is usually guided by estimations of their caloric needs. However, recent studies have shown that energy expenditure (EE) of critically ill patients is not as high as previously thought. The goal of this study was to measure EE in ICU patients, compare it with estimated EE and evaluate nutritional support. METHODS: Energy expenditure was measured with indirect calorimetry in a broad group of ICU patients requiring mechanical ventilation >48 hours. In comparison EE was estimated with the Harris Benedict equation. Nutritional support during ICU stay was registered. RESULTS: Mean measured EE of 56 patients was 1820 +/- 419 kcal/day or 22 kcal/kg/day. The Harris- Benedict equation underestimated EE by 11%, but adding a stress factor resulted in 15% overestimation. Mean nutritional support was 1175 +/- 442 kcal/day or 67% of EE. The energy deficit was greatest during the first week of ICU stay. Mean protein administration was 0,44 g/kg/day. CONCLUSION: Measured EE of ICU patients was less than nutritional support recommended by international guidelines. These results are in accordance with recent studies. Nutritional support was only 67% of measured energy expenditure and protein content less than recommended. Further studies are needed as it has not be shown how this might influence outcome. PMID- 19553708 TI - [Multidrug resistant tuberculosis in Iceland--case series and review of the literature]. AB - BACKGROUND: Multidrug resistant tuberculosis (MDR-TB) is a growing health problem in the world. Treatment outcomes are poorer, duration longer and costs higher. We report three cases of MDR-TB diagnosed in Iceland in a six year period, 2003-8. CASE DESCRIPTIONS: The first case was a 23-year-old immigrant with a prior history of latent TB infection treated with isoniazid. He was admitted two years later with peritoneal MDR-TB. He was treated for 18 months and improved. The second case was a 23-year-old immigrant diagnosed with pulmonary MDR-TB after having dropped out of treatment in his country of origin. Clinical and microbiological response was achieved and two years of treatment were planned. The third case involved a 27-year-old asymptomatic woman diagnosed with MDR-TB on contact tracing, because of her brother's MDR-TB. 18 months of treatment were planned. CONCLUSIONS: Clustering of cases of MDR-TB in the last six years, accounting for almost 5% of all Icelandic TB cases in the period, suggests that an increase in incidence might be seen in Iceland in coming years. The infection poses a health risk to the patients and the general public as well as a financial burden on the health care system. Emphasis should be put on rapid diagnosis and correct treatment, together with appropriate immigration screening and contact tracing. PMID- 19553709 TI - [In-hospital cardiopulmonary resuscitation at Landspitali University Hospital in Reykjavik]. AB - INTRODUCTION: Survival after in-hospital cardiac arrest has not been previously reported in Iceland and the purpose of this study was to examine the outcomes of in-hospital resuscitation over a two year period. MATERIAL AND METHODS: There are resuscitation teams on each of the two campuses of the University Hospital in Reykjavik. Since the beginning of 2006, the resuscitation teams have compiled their reports in a structured form, Utstein style. RESULTS: During 2006 and 2007 resuscitation teams were activated on a total of 311 occasions. Of those, there was need for a full cardiopulmonary resuscitation because of cardiac arrest of in patients in 80 cases (26%). Return of spontaneous circulation was achieved or the patient survived to be transferred to the intensive care unit in 55 (69%) of the 80 cases. Survival to discharge was 33%. Survival to discharge was better if the arrest occurred between 8 AM and 4 PM during daytime (50%), than outside of regular working hours (23%, p=0.02). The survival was better if ventricular tachycardia or fibrillation was the first rhythm encountered (50%) than if the initial rhythm turned out to be asystole or pulseless electrical activity (12%, p=0.002). Those who survived resuscitation were generally younger than those who did not (p=0.002). CONCLUSION: The outcomes were similar to those reported at institutions in our neighboring countries. The survival rate was lower if the cardiac arrest occurred outside of regular working hours and if ventricular tachycardia or fibrillation was the first encountered rhythm. PMID- 19553710 TI - Vision and challenges for dental research worker. PMID- 19553711 TI - The relative diagnostic yields of clinical, FOTI and radiographic examinations for the detection of approximal caries in youngsters. AB - BACKGROUND: The detection of carious lesions in the initial stages of development is very important in order to prevent the occurrence of cavitation. AIMS AND OBJECTIVES: The aim of the present study was to compare three methods-visual examination, FOTI, and bitewing radiographic examination-for the detection of noncavitated and cavitated approximal carious lesions. MATERIALS AND METHODS: Seventy students (mean age 14 years) with low caries prevalence were examined by three examiners. The tooth surfaces judged as having caries by at least one examiner or one diagnostic method were scheduled for tooth separation. RESULTS: The results showed that the incorporation of FOTI and radiographic examination represented an additional diagnostic yield of 50% and about 110%, respectively, compared to clinical examination alone. CONCLUSION: We conclude that FOTI or radiographic examination, or both, used as adjuncts to clinical examination, could improve the detection of noncavitated and cavitated approximal carious lesions. Although FOTI should not replace bitewing radiographic examination, it does seem to have additional value for the detection of carious lesions on approximal surfaces of the posterior teeth in school children with low caries prevalence. PMID- 19553712 TI - Porphyromonas gingivalis decreases osteoblast proliferation through IL-6 RANKL/OPG and MMP-9/TIMPs pathways. AB - BACKGROUND: Porphyromonas gingivalis, an important periodontal pathogen, is closely associated with inflammatory alveolar bone resorption. This bacterium exerts its pathogenic effect indirectly through multiple virulence factors, such as lipopolysaccharides, fimbriae, and proteases. Another possible pathogenic path may be through a direct interaction with the host's soft and hard tissues (e.g., alveolar bone), which could lead to periodontitis. AIMS AND OBJECTIVES: The aim of the present study was to investigate the direct effect of live and heat inactivated P gingivalis on bone resorption, using an in vitro osteoblast culture model. RESULTS: Optical microscopy and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide MTT assay revealed that live P gingivalis induced osteoblast detachment and reduced their proliferation. This effect was specific to live bacteria and was dependent on their concentration. Live P gingivalis increased IL 6 mRNA expression and protein production and downregulated RANKL and OPG mRNA expression. The effect of live P gingivalis on bone resorption was strengthened by an increase in MMP-9 expression and its activity. This increase was accompanied by an increase in TIMP-1 and TIMP-2 mRNA expression and protein production by osteoblasts infected with live P gingivalis. CONCLUSION: Overall, the results suggest that direct contact of P gingivalis with osteoblasts induces bone resorption through an inflammatory pathway that involves IL-6, RANKL/OPG, and MMP-9/TIMPs. PMID- 19553713 TI - Radiographic evaluation of the mental foramen in a selected Iranian population. AB - BACKGROUND: Information on the position of the mental foramen is important for dental surgeons. Variations in its position can be a cause of complications during local anesthesia or surgical procedures. The usual position of the mental foramen in an Iranian population has not been previously reported. AIMS: The purpose of this study was to determine the most common location of the mental foramen in an Iranian population. We also analyzed gender differences and the symmetry of location within individuals. MATERIALS AND METHODS: 400 panoramic radiographs were evaluated with regard to the location and symmetry of the mental foramina in male and female subjects. RESULTS: We found that the mental foramen was located between the first and second premolars in 47.2% of patients and in line with the second premolar in 46%. In 49.2% of males, the mental foramen was in line with the second premolar. In 50.9% of females it was between the first and second premolars. It was symmetrical in 85.7%. CONCLUSIONS: Based on this study it appears that the most common position of mental foramen is either between the two premolars or in line with the second premolar. This is in concordance with previous studies. PMID- 19553714 TI - Influence of different post design and composition on stress distribution in maxillary central incisor: Finite element analysis. AB - BACKGROUND: Post design and material has very important effects on dentinal stress distribution since the post placement can create stresses that lead to root fracture. MATERIALS AND METHODS: In this study we use finite element analysis (FEA) to evaluate stress distribution on endodontically treated maxillary central incisors that have been restored with different prefabricated posts. Six models were generated from the image of anatomical plate: Four metallic posts (ParaPost XH, ParaPost XT, ParaPost XP, and Flexi-Flange) and one fiberglass post (ParaPost Fiber Lux). The sixth model was a control-a sound maxillary central incisor. We used CAD software and exported the models to ANSYS 9.0. All the materials and structures were considered elastic, isotropic, homogeneous, and linear except the fiberglass post which was considered orthotropic. The values for the mechanical properties were obtained by a review of the literature and the model was meshed with 8-node tetrahedral elements. A load of 2N was applied to the lingual surface at an angle of 135 degrees. RESULTS: The stress results were recorded by shear stress and von Mises criteria; it was observed that there was no difference for stress distribution among the titanium posts in the radicular portions and into posts. There was higher stress concentration on the coronary portion with the titanium posts than with the glass fiber post. It seems that the metallic posts' external configuration does not influence the stress distribution. CONCLUSION: Fiber posts show more homogeneous stress distribution than metallic posts. The post material seems to be more relevant for the stress distribution in endodontically treated teeth than the posts' external configuration. PMID- 19553715 TI - Influence of flowable materials on microleakage of nanofilled and hybrid Class II composite restorations with LED and QTH LCUs. AB - BACKGROUND: Class II composite restorations are more frequently being placed with margins apical to the cementoenamel junction (CEJ) and margins within the dentin are prone to microleakage. AIMS: This in vitro study was used to evaluate the influence of flowable composite and flowable compomer as gingival liner on microleakage in Class II composite restorations and compare a light-emitting diode (LED) unit with a quartz tungsten halogen (QTH) unit for light-activating composite resins. MATERIALS AND METHODS: Mesioocclusal and distoocclusal Class II cavity preparations were made in 72 sound extracted premolars. The buccolingual width was 2.5 mm and the gingival margins of all the cavities were placed 1.0 mm apical to the CEJ. The boxes were prepared 1.5 mm deep axially, making 144 slot cavities. Teeth were randomly divided into the following two groups (n = 72): (I) Universal Filtek Supreme XT; Universal Filtek Supreme XT + Flwable Filtek XT and Universal Filtek Supreme XT + Dyract Flow and (II) Filtek Z250; Filtek Z250 + Flwable Filtek XT and Filtek Z250 + Dyract Flow. Flowable materials were injected into the gingival floor of the cavity to a thickness of 1.0 mm. Each increment was cured for 20 s. One-half of the subgroups in each group were cured with QTH and the other half with LED light curing units (LCUs). After 1 week of incubation at 37 degrees C, the specimens were thermocycled (5-55 degrees C, x1500), immersed in 0.5% basic fuchsine dye for 24 h and sectioned and microleakage was evaluated at the gingival margin by two examiners using a 0-3 score scale. The data were analyzed using the Kruskal-Wallis and Mann-Whitney U tests. RESULTS: The groups utilizing flowable liners had significantly less microleakage (P < 0.05). No significant difference was identified between Universal Filtek Supreme XT and Filtek Z250 composites with and without flowable materials. There was no significant between utilizing flowable composite or flowable compomer and between each similar subgroup when polymerized with either the LED or the QTH LCUs. CONCLUSIONS: A layer of flowable materials at the gingival floor of Class II composite restorations may be recommended to improve the marginal seal of a restoration. PMID- 19553716 TI - The knowledge and attitude of general dentists toward denture adhesives in Tehran. AB - BACKGROUND: The purpose of this study was to generate discussion and communication among a group of general dentists in Tehran on their viewpoints regarding denture adhesives. Have they accepted denture adhesive as a material to enhance denture retention, stability and function? MATERIALS AND METHODS: In the summer of 2007, a questionnaire was mailed to 300 general dentists who were assigned with a random systematic sampling method from general dentists in Tehran. The questions were arranged in two parts of evaluating knowledge and attitude. In evaluation of knowledge, dentists were classified into groups of good, moderate, weak and lack of knowledge. In evaluation of attitude, dentists were classified into positive, moderate and negative groups. (Evaluating attitude was carried out in good and moderate groups of knowledge.) RESULTS: The study showed that 14%, 32% and 37% of the general dentists had respectively good, moderate and weak knowledge toward denture adhesive while 16.3% had no knowledge about this material. In evaluation of attitude through dentists with positive and moderate knowledge toward denture adhesive, 9.3%, 71.3% and 19.4% had respectively positive, moderate and negative attitude toward denture adhesive. The chi2 test showed a significant statistical relation between situation of knowledge and experiences of dentists. DISCUSSION: This study demonstrated that the rate of knowledge of these 300 general dentists in Tehran towards denture adhesives has not been in a good situation. It is believed that denture adhesive be able to enhance the fitness of a denture and provide psychological relief to the patient. Dentists agreed that education, not only for practitioners but also for patients, would raise the advantageous features and reduce the misuse of denture adhesive. Education of the topic "Denture adhesive" should be more concerned in dental universities. PMID- 19553717 TI - Cytotoxicity evaluation of Persica mouthwash on cultured human and mouse cell lines in the presence and absence of fetal calf serum. AB - BACKGROUND AND AIMS: The effectiveness of an ideal antimicrobial agent depends on its ability to kill microbes while causing minimal toxicity to host cells. Several studies have been reported on the antimicrobial effects of chewing sticks (Salvadora persica) on oral bacteria. The purpose of this study was to evaluate the cytotoxic effects of Persica and chlorhexidine (CHX) mouthwashes on cultured human and mouse cell lines. MATERIALS AND METHODS: This was an experimental study. The toxic effects of four dilutions of Persica and CHX mouthwashes on KB, Saos-2, J744 A1, and gingival fibroblast cells were evaluated by MTT [3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The effect of fetal calf serum (FCS) components on the cytotoxicity of these mouthwashes was also investigated. STATISTICAL ANALYSIS: Analysis of variance and the Kruskal Wallis test were used to evaluate the results. RESULTS: The results indicated that Persica , at concentrations higher than 0.1%, exerted a very significant cytotoxic effect on all the cell lines (P < or = 0.01). CHX, at a concentration of 0.001%, exerted toxic effects only on gingival fibroblasts; concentrations higher than 0.001% were required to produce significant cell death in the other cell lines. At all the concentrations under study, both Persica and CHX exerted significantly greater cytotoxic effects in the absence of FCS than in its presence (i.e., in control culture medium). The toxicities of both mouthwashes were attenuated in the presence of FCS (10%). CONCLUSION: Our results indicate that both Persica and CHX mouthwashes are toxic to macrophage, epithelial, fibroblast, and osteoblast cells in a concentration-dependent manner. PMID- 19553718 TI - Tensile bond strength of composite luting cements to metal alloys after various surface treatments. AB - AIMS: To evaluate the effects of two different surface treatments and bonding agents on tensile bond strength between a Co-Cr and a Ni-Cr cast alloy and two resin-luting cements. MATERIALS AND METHODS: Two hundred and forty alloy samples were cast and subjected to surface treatments such as sandblasting, chemical etching, and sandblasting plus chemical etching. Panavia F and CandB cement were used as cementing mediums. The etching qualities were examined by a stereooptic microscope. Failure surfaces were examined throughout scanning electron microscopy. The data were evaluated using statistical methods, namely analysis of variance and multiple comparison test (Tukey HSD). RESULTS: Significant differences were found in the bonding provided by the various cements (P < 0.001) and also type of surface treatments (P < 0.001). For all groups, sandblasted surfaces showed the highest bond strength values. There was no significant difference between the Cr-Co and the Cr-Ni alloys (P > 0.05). CONCLUSIONS: Panavia F showed higher tensile strength and the sandblasted samples possessed higher tensile strength. PMID- 19553719 TI - Effect of repeated use on dentin bond strength of two adhesive systems: all-in one and one-bottle. AB - AIMS: To compare the effects of repeated use of two one-bottle adhesives with that of two all- in- one adhesives (with acetone solvent) on bond strength to dentin. MATERIALS AND METHODS: A flat dentin surface was prepared on 120 bovine incisors using 600- grit abrasive pape. The teeth were randomly assigned into 12 equal groups. The four adhesive systems [Prime and Bond NT (P&B NT), One-Step Plus (OS), iBond (iB), and G-Bond (GB)] were used at baseline, after the lid of the container had been opened 30 times, and after it had been opened 60 times. Before each use of the adhesives, the lids of the containers were left open for 1 min. The resin composites were applied on the dentin in a cylindrical split mold. After thermocycling, shear bond strength test was performed with a universal testing machine at 1 mm/min. We used Kruskal-Wallis and Dunn tests for statistical analysis. RESULTS: There was no statistically significant difference among bond strength (MPa) of the groups of P&B NT (31.9 +/- 4.6, 31.8 +/- 6.5, 26.1 +/- 6.7) and OS (33.2 +/- 5.1, 30.9 +/- 7, 29.3 +/- 5.9), respectively (P > 0.05). The mean of the bond strength of iB and GB after 60 times (15.3 +/- 4.1 and 12.2 +/- 3.9, respectively) was significantly lower than that of iB and GB at baseline (23.5 +/- 4.8 and 22.2 +/- 4.5, respectively) (P < 0.05). CONCLUSIONS: Repeated use (60 times) of the all-in-one adhesive led to a decline in the dentin bond strength. To avoid this problem it would be advisable to have containers with smaller amounts of adhesive or perhaps those with only a singe dose. PMID- 19553720 TI - The influence of water temperature during toothbrushing on root dentine: an in vitro study. AB - BACKGROUND/AIMS: The use of cold water during toothbrushing can cause dentine sensitivity and, to avoid this painful stimulus, some patients used to rinse their mouths with warm water when brushing. Thus, the objective of this study was to determine the effect of water temperature on the dental root surface during toothbrushing. MATERIALS AND METHODS: Fragments of bovine dental roots were submitted to 15,000 strokes in a toothbrushing machine using a slurry of toothpaste/water, medium brushes, and a 200-g load. They were randomly divided into two groups: toothbrushing with cold water or with hot water. Tooth wear was measured by loss of weight and by tissue height. STATISTICAL ANALYSIS: The weight and height data obtained in 17 and 10 replicas respectively are presented as mean +/- standard error of mean. The data were compared using the Kolgomorov-Smirnof (Lilliefors) test followed by one-way ANOVA. The level of significance was 5% (P < 0.05). RESULTS: There were no significant differences between the two experimental groups. The mean percentages of weight losses were 5.61 +/- 1.66 for cold water and 6.25 +/- 1.98 for hot water. The mean dentine height losses were 51.02 +/- 15.92 microm for cold water and 63.54 +/- 17.75 microm for hot water. CONCLUSION: The use of warm water during toothbrushing promoted root dentine wear similar to that produced by the use of cold water. The results suggest that warm or cold water may be used during toothbrushing without any additional damage to the patients' dental hard tissues. PMID- 19553721 TI - Association between symptoms of temporomandibular disorders and gender, morphological occlusion, and psychological factors in a group of university students. AB - AIM: The purpose of this study was to find out the prevalence of temporomandibular disorder (TMD) in a sample of university students and its relationship to gender, occlusion, and psychological factors. MATERIALS AND METHODS: The sample comprised 196 subjects, aged 18-25 years. The TMD degree was evaluated using an anamnestic questionnaire. Morphologic occlusion was evaluated according to Angle classification (classes I, II, and III). The Hospital Anxiety and Depression Scale (HADS), a 14-item self-administered rating scale developed specifically to identify anxiety and depression in nonpsychiatric medical outpatients, was used to assess the levels of anxiety (HADSa) and depression (HADSd). STATISTICAL ANALYSIS: The incidence of TMD level, malocclusion, anxiety, and depression in both genders was calculated as percentages. Association between TMD degree and occlusion, HADSa, and HADSd was tested using the Chi-square test. RESULTS: According to our results, 50% of the subjects had TMD, but it was of moderate or severe degree in only 9.18% of them. No statistically significant association could be found between TMD and gender or occlusion. TMD was found to have statistically significant association with HADSa but not with HADSd. CONCLUSION: A high prevalence of TMD was found in this student population; however, most of the cases could be classified as mild. Of the variables studied, only HADSa had a statistically significant association with TMD. PMID- 19553722 TI - Dental esthetic satisfaction, received and desired dental treatments for improvement of esthetics. AB - AIM: The purposes of this research were to investigate factors influencing patients' satisfaction with their present dental esthetic, received previous dental treatments on anterior teeth and basic treatments that they wanted to undergo to improve their dental appearance. MATERIALS AND METHODS: A total of 1014 patients who attended a dental school in a major city in Turkey participated in the study. The participants were surveyed with a questionnaire containing questions about gender, age, education level, self-reported tooth appearance, received previous dental treatments on anterior teeth and desired basic esthetic dental treatments. Statistical analysis of the verifying data was made with descriptive statistics, chi2 test and multiple logistic regression analyses. RESULTS: According to the analyses of the verifying data, 55.1% of the patients were dissatisfied with the color of their teeth, 42.7% with dental appearance, 29.9% with crowding of anterior teeth, 23.3% were hiding teeth while smiling, 16.1% had non-esthetic restorations and 11.9% thought that their anterior teeth were protruding. Esthetic restoration was found to be the most-performed treatment recently (29.0%) and whitening of teeth was the most-desired dental treatment (49.0%). Gender, age and education level had an effect on satisfaction and received previous and desired dental treatments for improvement of esthetics. CONCLUSION: Many of the Turkish patients surveyed in the study were dissatisfied and desired the improvement of dental esthetics. Therefore, dentists should consider this as an important dimension in their practice. PMID- 19553723 TI - The relationship between overjet size and dentoalveolar compensation. AB - BACKGROUND: The purpose of this study was to investigate the effect of overjet size and the dento-alveolar compensation in subjects with normal class I molar relationship. MATERIALS AND METHODS: Lateral cephalometric head records of 59 Egyptian children (34 boys and 25 girls) aged 7.5 to 10.5 years with mean age of 8.69+/-0.73. All had normal class I type of occlusion. The sample was classified into four quartiles according to the overjet size and the cephalometric analysis was based on seven linear and eight angular measurements using a dental tracer programme. RESULTS: showed that, in spite of presence of high significant over jet size differences between the groups; there was no significant differences in all the studied parameters were found. Applying the least significant differences (LSD) test and coefficient correlations between the studied parameters clarifying that there was a significant differences in angular measurements (SN-AB, SN-Occl, I-I, I-ML, I-NB). CONCLUSION: during transitional dentition there was a sufficient dento-alveolar adaptation to growth changes in the saggittal jaw relation ship to attain normal class I type of occlusion. This compensation is pronounced in angular parameters and clustered in the lower arches particularly in incisal area. PMID- 19553724 TI - Helicobacter pylori coinfection is a confounder, modulating mucosal inflammation in oral submucous fibrosis. AB - The oral cavity has been considered a potential reservoir for Helicobacter pylori (H pylori) , from where the organism causes recurrent gastric infections. AIM: With this case-control study we tried to evaluate the role of H pylori in the etiology of mucosal inflammation, a condition that compounds the morbid state associated with oral submucous fibrosis (OSF). MATERIALS AND METHODS: Subjects ( n = 150) were selected following institutional regulations on sample collection and grouped into test cases and positive and negative controls based on the presence of mucosal fibrosis and inflammation. The negative controls had none of the clinical signs. All patients underwent an oral examination as well as tests to assess oral hygiene/periodontal disease status; a rapid urease test (RUT) of plaque samples was also done to estimate the H pylori bacterial load. We used univariate and mutivariate logistic regression for statistical analysis of the data and calculated the odds ratios to assess the risk posed by the different variables. RESULTS: The RUT results differed significantly between the groups, reflecting the variations in the bacterial loads in each category. The test was positive in 52% in the positive controls (where nonspecific inflammation of oral mucosa was seen unassociated with fibrosis), in 46% of the test cases, and in 18% of the negative controls (healthy volunteers) (chi2 = 13.887; P < 0.01). A positive correlation was seen between the oral hygiene/periodontal disease indices and RUT reactivity in all the three groups. CONCLUSIONS: The contribution of the H pylori in dental plaque to mucosal inflammation and periodontal disease was significant. Logistic regression analysis showed gastrointestinal disease and poor oral hygiene as being the greatest risk factors for bacterial colonization, irrespective of the subject groups. A positive correlation exists between RUT reactivity and the frequency of mucosal inflammation. PMID- 19553725 TI - Etiopathogenesis of disk displacement of the temporomandibular joint: a review of the mechanisms. AB - Disk displacement of the temporomandibular joint (TMJ) is a clinically important condition, showing a high prevalence in both patient and non-patient populations. Despite its clinical importance, there is incomplete understanding of the etiopathogenic mechanisms leading to disk displacement. A number of possible risk factors have been identified. This article analyzes the etiopathogenesis from both the clinical and the biomechanical viewpoints and also reviews the literature on the association between disk displacement and the main risk factors (i.e., trauma, altered disk shape and/or dynamic properties, occlusal abnormalities, steepness of the articular eminence, hyperactivity of the lateral pterygoid muscle, joint hypermobility, etc.). According to our interpretation of available data, an impairment of joint lubrication may be a common finding in cases of disk displacement, thus suggesting the need for future studies addressing both local and systemic neuroendocrine aspects influencing the friction coefficient of the TMJ. A full comprehension of the etiopathogenesis of disk displacement is far from being achieved, and clinicians must take into account this consideration when treating patients with temporomandibular disorders. PMID- 19553727 TI - Riga-Fede disease: a histological study and case report. AB - Acute traumatic ulcerations and granulomas of the oral mucosa may result from physical damage via sharp foodstuffs, accidental biting, or talking. Most ulcerations heal within days. Others become chronic, reactive, and exophytic. A histopathologically unique type of chronic traumatic ulceration is the traumatic ulcerative granuloma with stromal eosinophilia (TUGSE). TUGSE exhibits a deep "pseudoinvasive" inflammatory reaction. This lesion may occur under the tongue in infants as a result of chronic mucosal trauma caused by mandibular anterior primary teeth during nursing and is termed Riga-Fede disease (RFD). The clinical presentation many resemble squamous cell carcinoma causing concern. RFD, although not uncommon, is not frequently reported. Thus, dental practitioners are unfamiliar with such lesions. We present a large Riga-Fede lesion in an infant along with the clinical management. PMID- 19553726 TI - Tissue engineering and its implications in dentistry. AB - Tissue engineering is a novel and highly exciting field of research. With tissue engineering techniques it may be possible to repair damaged tissues or even create replacement organs. This article reviews the principles underlying key tissue engineering strategies and the typical components used. Examples of tissue engineering include passive approaches, such as dental implants, and inductive approaches, in which specific molecular signals are used to activate cells. PMID- 19553728 TI - Odontoameloblastoma: report of two cases. AB - Odontoameloblastoma (OA) is a very rare mixed odontogenic neoplasm, characterized by the simultaneous occurrence of an ameloblastoma and a compound or complex odontoma in the same tumor mass. To date, less than 50 cases of OA and/or ameloblastic odontoma have been reported in the English dental literature. This neoplasm was called ameloblastic odontoma. The term OA was included in the 1971 WHO classification. In this study, we present two cases of OA, which we hope will contribute to the awareness and knowledge of surgeons regarding the existence of this odontogenic tumor so that patients having it may be treated and followed-up properly. PMID- 19553729 TI - Bilateral bifid mandibular canal: report of two cases. AB - Bifid mandibular canal is a rare anatomical variation that can be of considerable interest to a dentist. This condition can lead to complications when performing mandibular anesthesia or during surgery of the lower third molar, orthognatic or reconstructive mandibular surgery, or placement of dental implants and prosthesis; bleeding and traumatic neuroma are possible complications. Therefore, awareness of this condition is important. We report two cases of bilateral bifid mandibular canal: one in a 22-year-old male and the other in a 24-year-old female. PMID- 19553730 TI - Bilateral dens invaginatus in the mandibular premolar region. AB - Dens invaginatus (dens in dente, DI) is a rare developmental anomaly resulting from invagination of a portion of the crown rare. It is an important dental anomaly due to the possible pulpal involvement. DI can be detected clinically in a tooth presenting unusual crown morphology or having deep foramen coaceum. Also, clinically, non-suspected affected teeth are commonly diagnosed as an incidental radiographic finding. Presence of DI in the mandibular premolar teeth is unusual. Aims is to introduce a case of bilateral occurrence of DI in mandibular first premolar teeth. A 33- year-old man was referred to our clinic with a chief complaint of severe pain in his lower third molar tooth. Radiographic examination revealed bilateral DI (single at the left, double at the right) in mandibular first premolar teeth. The teeth were restored with fissure sealant. Although bilateral appearance of DI is a frequent situation, mandibular occurrence is very rare. Our review of the literature reveals just nine cases of DI, and only one of them is in a premolar tooth. PMID- 19553731 TI - Three root canals in the maxillary second premolar. AB - In this study, we report an endodontic treatment of the maxillary second premolar with three root canals and distinct foramens. The possibility of three root canals in this tooth is quite small; however, it must be taken into account in clinical and radiographic evaluation during endodontic treatment. Many times, their presence is noticed only after canal treatment due to continuing post operative discomfort. PMID- 19553732 TI - Endodontic treatment of a periradicular lesion on an invaginated type III mandibular lateral incisor. AB - Dens invaginatus (DI), commonly known as dens in dente, is a developmental malformation of teeth that most commonly affects permanent maxillary incisor teeth. DI can present in a variety of forms, knowledge of which can usefully help in endodontic diagnosis and treatment. This article reports on an unusual case of DI type III with a periradicular lesion in a mandibular lateral incisor. Non surgical endodontic treatment was performed and resolution of the periradicular lesion was observed at 1 year follow-up. Clinical considerations and treatment are discussed and reported. PMID- 19553733 TI - How useful is journal impact factor? PMID- 19553734 TI - Use of Ayurveda in promoting dental health and preventing dental caries. PMID- 19553735 TI - Porphyromonas gingivalis and Porphyromonas endodontalis and their roles in systematic diseases: true or false? PMID- 19553736 TI - Open questions on carcinogenesis of oral cancer: interaction between the environmental and genetic aspects. PMID- 19553737 TI - [Age at death in the Turkish Adult Risk Factor Study: temporal trend and regional distribution at 56,700 person-years' follow-up]. AB - OBJECTIVES: We analyzed the temporal trend and regional distribution of age at all-cause death and the sex-specific and age-bracket defined coronary mortality in the 18-year follow-up of the Turkish Adult Risk Factor Study. STUDY DESIGN: The participants of the Turkish Adult Risk Factor Study who have been examined in even years were last surveyed in August 2008. A total of 1,582 individuals were surveyed, which constituted half of the alive participants of the overall cohort. Information on death was obtained from first-degree relatives and/or health personnel of local heath offices. Survivors were evaluated by history, physical examination, and 12-lead electrocardiography. The cumulative follow-up was 56,700 person-years. RESULTS: Of 1,582 participants, 868 (431 men, 437 women) were examined, in 604 subjects information was gathered, and 47 participants (26 men, 21 women) were ascertained to have died. Twenty-two deaths were classified as of coronary origin. Cumulative assessment of the entire cohort in the age bracket of 45-74 years disclosed coronary mortality to be 7.64 per 1000 person-years in men and 3.84 in women and persisted to be the highest among 30 European countries, whereas overall mortality declined at a greater proportion. Overall mean ages at death were deferred within a 12-year period by 7.4 years in men and 6 years in women, to 71.9 and 74.8 years, respectively. The extension of this mean survival was similar among urban-rural areas and geographic regions. CONCLUSION: Coronary mortality declined modestly, but life expectancy of Turkish adults rose by a mean of nearly seven years in the 12 years to 2003-08, without showing major differences in sex, urban-rural dwelling, or geographic regions. PMID- 19553738 TI - [Oral anticoagulant use in patients with atrial fibrillation]. AB - OBJECTIVES: Oral anticoagulant therapy has been shown to decrease the risk for vascular complications in patients with atrial fibrillation (AF). We evaluated the frequency of oral anticoagulant use in patients with AF, whether oral anticoagulant use was associated with effective INR values, and the reasons for not including an anticoagulant in the treatment. STUDY DESIGN: The study included 426 consecutive patients (256 women, 170 men; mean age 66+/-11 years) who presented with a diagnosis of AF between October 2007 and November 2008. The patients were inquired about whether they were using warfarin and/or aspirin and the reasons for not taking an oral anticoagulant. The INR levels were measured in those receiving warfarin. RESULTS: Permanent AF was present in 72.8%, and paroxysmal AF was present in 27.2%. Patients = or >75 years of age accounted for 32.4%. The risk for stroke was high in 69.3%, moderate in 21.8%, and low in 8.9%, hypertension being the most frequent risk factor (66.7%). Inquiry about medications showed that 107 patients (25.1%) were taking aspirin and warfarin, 21 patients (4.9%) and 237 patients (55.6%) were taking warfarin and aspirin alone, respectively, while 61 patients (14.3%) used none. The incidence of oral anticoagulant use was 30.1%, being significantly low in patients = or >75 years of age (p=0.0001), and having hypertension (p=0.023) or coronary artery disease (p=0.004). Effective INR values recommended by the guidelines were attained in 47.7% (n=61) of patients receiving warfarin. Sex, age, clinical risk factors, and socioeconomic parameters were not associated with achievement of target INR values. The most frequent reason for not starting anticoagulant treatment was the low tendency of physicians to prescribe the drug (74.3%), followed by the presence of contraindications (9.8%). CONCLUSION: The most important factor for inadequate oral anticoagulant use especially in patients having a high risk for stroke is the low incidence of prescription of the drug by the physicians, suggesting low influence of the guidelines on the clinical practice. PMID- 19553739 TI - Relationship between elevated serum gamma-glutamyltransferase activity and slow coronary flow. AB - OBJECTIVES: We evaluated the relationship between coronary blood flow and serum gamma-glutamyltransferase (GGT) activity in patients with slow coronary flow (SCF). STUDY DESIGN: The study included 90 patients (47 men, 43 women; mean age 50.8+/-9.4 years) with SCF and 88 patients (45 men, 43 women; mean age 51.4+/-8.8 years) with coronary artery disease (CAD), whose diagnoses were made by coronary angiography. Patients with CAD had normal coronary flow. Coronary flow was quantified using the corrected TIMI frame count (TFC) method and serum levels of gamma-glutamyltransferase were measured. The results were compared with those of a control group consisting of 86 age- and sex-matched patients who had normal coronary arteries and normal coronary flow. RESULTS: The three groups were similar with respect to body mass index, presence of hypertension and diabetes mellitus, lipid profiles, and fasting glucose. The use of medications was significantly more common in the CAD group (p<0.01). Compared to the control group, serum GGT activity was significantly increased in both SCF and CAD groups (p<0.01), but these two groups did not differ significantly in this respect (p=0.71). The TFCs for all the epicardial coronary arteries and the mean TFC were significantly higher in the SCF group (p<0.01). Patients with CAD and the controls had similar TFC parameters. The mean TFC showed a positive and moderate correlation with serum GGT activity (r=0.326; p<0.001). In regression analysis, serum GGT activity was found as the only independent predictor of the mean TFC (beta=0.309; p<0.001). CONCLUSION: We have shown for the first time an association between increased serum GGT activity and SCF. Further clinical studies are needed to clarify the physiopathologic role of serum GGT activity in SCF. PMID- 19553740 TI - [Slow coronary flow]. PMID- 19553741 TI - [Echocardiography-guided pericardiocentesis with the apical approach]. AB - OBJECTIVES: We aimed to evaluate our experience with echocardiography-guided pericardiocentesis with the apical approach for pericardial effusions. STUDY DESIGN: We evaluated 32 pericardiocenteses performed under echocardiography guidance and with the apical approach in 29 patients (15 men, 14 women; mean age 49 years; range 18 to 72 years). Indications were diagnostic purpose, pericardial tamponade, or symptomatic pericardial effusion. Procedural success, the amount of drainage, and complications were assessed. RESULTS: Common causes of pericardial effusion were malignancy (n=6), postpericardiotomy syndrome (n=5), idiopathic (n=5), chronic renal disease (n=4), and myocardial infarction (n=3). The amount of drainage was 120 ml to 2,200 ml and the duration of pericardial catheter placement in the pericardial space was 24 to 144 hours. Mortality did not occur. Echocardiographic control showed residual effusion in the lateral wall in one case, which required repositioning of the pericardial catheter for complete removal. The procedure failed in one patient due to insufficient drainage caused by multiple septations and fibrinous fluid in the pericardial space. The success rate of the procedures was 96.9%. Four cases developed hemopneumothorax requiring tube drainage, vasovagal reaction, nonsustained ventricular tachycardia, and frequent ventricular extrasystoles, respectively. Apical puncture was repeated in two cases due to erroneous left ventricular puncture and pleural catheter placement, respectively. CONCLUSION: Echocardiography-guided pericardiocentesis with the apical approach is readily performed bedside without the need for catheterization laboratory, with a high success rate and low complication rate. It should be considered especially in cases in which anterior pericardial collection is more prominent where it will reduce unnecessary surgical interventions. PMID- 19553742 TI - [Clinical and epidemiological characteristics of infective endocarditis]. AB - OBJECTIVES: We evaluated the clinical and laboratory characteristics of patients with infective endocarditis (IE). STUDY DESIGN: During a five-year period (January 2002-December 2006), 96 patients (56 men, 40 women; mean age 47+/-15 years; range 16 to 81 years) were treated and followed for IE. All the patients underwent transthoracic and transesophageal echocardiography. Clinical and laboratory characteristics of the patients were reviewed. All cases of prosthetic valve endocarditis were late-onset IE. Blood samples were cultured in both aerobic and anaerobic media and incubated for 21 days. The diagnosis of IE was made according to the Duke criteria. RESULTS: Male-to-female ratio was 1.4. The most common complaint on presentation was dyspnea (n=58, 60.4%), followed by fever (n=20, 20.8%), and fatigue (n=16, 16.7%). The mean follow-up period was 21+/-12 days (range 2 to 52 days). While transthoracic echocardiography was sufficient to detect vegetations in 69 patients (71.9%), IE lesions could be demonstrated by transesophageal echocardiography in 27 patients (28.1%). The most common involvement was seen in the mitral valve (54.2%), and the incidence of native valve involvement (79.2%) was greater than that of prosthetic valve involvement. Forty-nine patients (51%) were submitted to surgery. In-hospital mortality occurred in 12 patients (12.5%). The prevalence of IE among all patients hospitalized in cardiology department was 0.13%. The culprit microorganism was isolated in only 36 patients (37.5%), being staphylococci (33.3%), enterococci (27.8%), brucellae (22.2%), and streptococci (11.1%). CONCLUSION: The diagnosis and treatment of IE should be designed taking into consideration that epidemiologic and clinical features of IE may differ from those of individual countries. PMID- 19553743 TI - Lipomatous hypertrophy of the interatrial septum: a case report. AB - Lipomatous hypertrophy of the interatrial septum is generally a benign disorder characterized by lipid accumulation in the interatrial septum. A 56-year-old asymptomatic woman with a history of hypertension and hyperlipidemia was referred to echocardiographic examination. Transthoracic echocardiography showed left ventricular hypertrophy, normal left ventricular systolic function, and left ventricular relaxation disturbance. The apical four-chamber view showed a hyperechogenic mass in the interatrial septum. Lipomatous hypertrophy was suspected and transesophageal echocardiography was performed. A dumbbell-shaped hypertrophy of 22 mm thickness was noted in the interatrial septum, which did not involve the fossa ovalis. There was no decrease in flow velocities of the superior and inferior vena cava nor a flow disturbance in the pulmonary veins. Cardiac magnetic resonance imaging showed lipomatosis and thickening in the interatrial septum and subepicardial region. PMID- 19553744 TI - Acute rupture of a congenital sinus of Valsalva aneurysm after severe exercise. AB - Ruptured aneurysms of the sinus of Valsalva may cause manifestations of prompt onset. A 22-year-old man with no previous cardiac complaints presented with dyspnea, palpitation, fatigue, and shortness of breath, all of which occurred after a football match. Transthoracic echocardiography showed an aneurysmal dilatation of the right sinus of Valsalva to the direction of the right ventricle. Color Doppler imaging showed a marked left-to-right turbulent flow from the aortic root to the right ventricle. Continuous Doppler imaging also revealed a continuous left-to-right systolodiastolic shunting. The patient was submitted to cardiovascular surgery for surgical repair. PMID- 19553745 TI - The contribution of MR coronary angiography to the diagnosis of a left anterior descending artery aneurysm in a patient with Kawasaki disease. AB - We present a 4-year-old boy with a diagnosis of Kawasaki disease of six-month history. At the time of diagnosis, no significant coronary artery aneurysm was detected by transthoracic echocardiography. On his routine follow-up echocardiography, dilatation of the left anterior descending (LAD) artery was suspected, but an exact diagnosis could not be made. He underwent magnetic resonance (MR) coronary angiography which showed a large fusiform aneurysmatic dilatation in the proximal segment of the LAD with a diameter of 9 mm. Coronary angiography performed to plan surgical repair confirmed the findings of MR angiography. The patient's parents refused surgical repair, so antiplatelet therapy was started to prevent thrombosis. No complications occurred during seven months of follow-up after detection of the aneurysm. PMID- 19553746 TI - Misleading ECG appearance of AV block due to concealed AV nodal conduction caused by interpolated ventricular ectopic beats. AB - Concealed conduction commonly occurs when a retrogradely conducted interpolated ectopic impulse enters the atrioventricular (AV) node; thus, the next sinus beat is not conducted to the ventricle or conducted with a prolonged PR interval because of increased refractoriness of AV conduction system. A 67-year-old man had complaints of exertional fatigue and palpitations at rest. His blood pressure was 110/70 mmHg and heart rate was 78 beats/min. Auscultation revealed a mild systolic murmur at the apex and an irregular rhythm. His electrocardiogram was normal, except for the presence of frequent premature ventricular complexes (PVC) of right bundle branch block morphology. Echocardiographic examination showed only grade-1 mitral regurgitation. Further evaluation with 24-h Holter monitoring showed frequent interpolated PVCs in bigeminal rhythm. Progressive prolongation of the PR interval was observed after each PVC, which ended with Mobitz type I AV block. The patient was treated with metoprolol which resulted in immediate and marked improvement in the symptoms. Control Holter recording showed very rare PVCs, without PR prolongation or AV block. PMID- 19553747 TI - [Implantation of the left ventricular pacemaker lead after successful balloon angioplasty for coronary vein stenosis: a report of two cases]. AB - Stenosis in the coronary veins can cause failure of left ventricular pacemaker lead implantation, which is the cornerstone of cardiac resynchronization therapy. There are several case reports in which left ventricular pacing could be possible after successful elimination of coronary vein stenosis by stent implantation or cutting balloon angioplasty. We report two cases of left ventricular pacemaker lead implantation after successful balloon angioplasty for posterolateral and posterior vein stenoses, respectively. PMID- 19553748 TI - [Myocardial noncompaction recognized following a transient ischemic attack]. AB - Myocardial noncompaction is a rare type of congenital cardiomyopathy characterized by excessively prominent trabeculations in one or more segments of the ventricles and deep intertrabecular recesses in ventricular walls. A 25-year old male patient presented to the neurology department with complaints of weakness in the left extremities. A mild loss of muscle strength was detected on neurological examination. With a preliminary diagnosis of acute cerebrovascular event, treatment with aspirin and enoxaparin was instituted, which improved his complaints within two hours. Electrocardiography showed sinus rhythm, left ventricular hypertrophy, and loss of R-wave progression in the precordial leads. Transthoracic echocardiography showed apical hypokinesia, marked left ventricular hypertrophy, and normal left ventricular diameters. There were numerous trabeculations in the apex, apical, lateral, and inferior walls, and deep intertrabecular recesses. Color Doppler showed blood flow into the intertrabecular recesses. He also had mild mitral regurgitation and diastolic dysfunction of restrictive type. He was scheduled for outpatient follow-up on aspirin and warfarin treatment. PMID- 19553749 TI - Prosthetic mitral valve thrombosis demonstrated by real-time 3D transesophageal echocardiography. PMID- 19553750 TI - [Demonstration of isolated persistent left superior vena cava by three dimensional multislice computed tomography]. PMID- 19553751 TI - [Pulmonary embolism as a rare complication of liposuction]. PMID- 19553752 TI - Huge intrapericardial aneurysm of the left atrial appendage. PMID- 19553753 TI - [Exercise-induced ventricular tachycardia associated with asymptomatic Brugada syndrome in a patient with urinary bladder stone]. PMID- 19553754 TI - [Prosthetic mechanic valve thrombosis in pregnant women]. PMID- 19553755 TI - [Atherosclerosis and the role of inflammation]. AB - Inflammation plays an important role in the development of atherosclerotic vascular disease which is the leading cause of morbidity and mortality in the adult population. Various risk factors trigger an inflammatory response leading to the initiation and development of atherosclerotic vascular disease. Several adhesion molecules, cytokines, and growth factors secreted from the endothelium aggravate inflammation and increase subendothelial lipid accumulation. With continuing inflammation, plaque rupture occurs resulting in acute coronary syndromes. Recently, the importance of the immune system and the mechanisms of the inflammatory response have been elucidated. One of the markers of subclinical inflammation in atherosclerosis is increased levels of high-sensitivity CRP. Several clinical trials have shown that suppression of the inflammatory response can delay or decrease the atherosclerotic process. Among the approaches for a healthier lifestyle, diets low in saturated fats, the Mediterranean diet, and exercise have been shown to decrease the inflammatory response. Various pharmacologic agents also have anti-inflammatory effects. Lipid-lowering therapies, in particular statins have proved to have anti-inflammatory effects on atheromatous plaques. Several clinical trials have shown that patients with an increased inflammatory response benefit more from statins. Therapeutic approaches targeting directly the inflammatory response and vaccination against atherosclerosis are under investigation. PMID- 19553756 TI - [The importance of non-lipid risk factors: a review focusing on C-reactive protein]. AB - Identification of individuals at risk before the development of atherosclerosis related diseases is important for preventive cardiology. Current risk calculators are not sufficient to effectively discriminate high-risk individuals from those having a low cardiovascular risk. It is obvious that new risk factors are needed to be used either alone or in conjunction with the current risk calculators to increase the accuracy of this discrimination. C-reactive protein (CRP) seems to be a good candidate as a biomarker, since it has been widely analyzed in prospective cohort studies and clinical trials. In this review, the value of CRP as a risk factor in atherosclerotic heart disease is discussed. PMID- 19553757 TI - [Primary prevention trials performed with statins prior to the JUPITER trial]. AB - The invention of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, namely statins, after the mid-1980s, initiated a new era in the approach to atherosclerotic diseases. Many studies of primary prevention, secondary prevention, and plaque regression have been carried out with statins. This paper will discuss primary prevention studies that were performed with statins prior to the JUPITER trial. PMID- 19553758 TI - [The JUPITER trial: a new approach in primary prevention]. AB - Current practice guidelines recommend that, when determining target LDL cholesterol levels in individuals without cardiovascular disease and diabetes, global risk estimation be made taking into account age, sex, total cholesterol level, HDL-cholesterol level, smoking status, and systolic blood pressure. Based on this assessment, target LDL-cholesterol levels have been set as <100 mg/dL, <130 mg/dL, and <160 mg/dL for high-, intermediate-, and low-risk individuals, respectively. The most important rationale for this approach is to implement a cost-effective treatment strategy, giving special attention to high-risk individuals when allocating available sources. Recent data suggest that increased levels of high sensitive C-reactive protein (hs-CRP) are associated with increased risk for cardiovascular events, independent of other risk parameters. Most recently, the JUPITER trial demonstrated that rosuvastatin 20 mg/day significantly lowered cardiovascular mortality and all-cause mortality in low- or intermediate-risk patients whose LDL-cholesterol levels were <130 mg/dL (median 108 mg/dL) and hs-CRP levels were =/>2 mg/L. As the current guidelines do not recommend statin therapy for this patient group, these results have caused a widespread interest, giving rise to arguments about the effectiveness of primary prevention recommendations of the guidelines. This review aims to discuss the results and implications of the JUPITER trial. PMID- 19553759 TI - It is always an organization's desire to improve and make adjustments to enhance services provided to others; the healthcare industry is no different. PMID- 19553760 TI - Politics, policy and practice: research for change in Canadian healthcare. PMID- 19553761 TI - Service-based funding and pay for performance: will incentive payments give Canadian healthcare the quality boost it needs? AB - "Pay fo performance" and "service-based funding" are fast-growing international trends, rapidly becoming dominant models in American and British healthcare. Now they're a bigger role in years to come. This reality led the Canadian Health Services Research foundation (CHSRF) to organize its third annual CEO forum on the subject in February 2009. PMID- 19553762 TI - Thoracic cancer surgeries. AB - In 2005, Cancer Care Ontario (CCO) released Thoracic Surgical Oncology Standards. These standards were aimed at providing the best level of care for those undergoing thoracic surgery and encompass surgeon training, hospital ancillary services and minimum volume thresholds for surgeries of the lung and esophagus. The objective of the current study was to explore variations in thoracic cancer surgical volumes at the hospital level across Canada. Using data from the Discharge Abstract Database for 2007-2008, the cohort included patients admitted to hospital with a most responsible diagnosis of cancer and who had a lung or esophageal surgery. To determine the volume of surgeries performed per facility, we grouped patients according to the hospital facility performing the surgery. In Canada (excluding Quebec and Prince Edward Island), there were a total 4,509 lung and 587 esophageal cancer procedures performed in 94 hospitals in 2007-2008. For both types of surgeries, Ontario hospitals performed approximately half of the procedures. Overall, 12 hospitals performed at or over the volume of surgeries for lung cancer as indentified by the CCO standards, while 10 did so for esophageal cancer. Nine hospitals performed both lung and esophageal cancer surgeries at or over the suggested volumes. Higher volumes of lung and esophageal cancer-related surgeries have been associated with improved patient outcomes. Here we present a snapshot of the distribution of cancer-related lung and esophageal surgeries across Canada (excluding Quebec and Prince Edward Island). PMID- 19553763 TI - New findings highlight potential risks of common drug combination in cardiac patients. PMID- 19553764 TI - Leading Lean: a Canadian healthcare leader's guide. AB - Canadian healthcare organizations are increasingly asked to do more with less, and too often this has resulted in demands on staff to simply work harder and longer. Lean methodologies, originating from Japanese industrial organizations and most notably Toyota, offer an alternative - tried and tested approaches to working smarter. Lean, with its systematic approaches to reducing waste, has found its way to Canadian healthcare organizations with promising results. This article reports on a study of five Canadian healthcare providers that have recently implemented Lean. We offer stories of success but also identify potential obstacles and ways by which they may be surmounted to provide better value for our healthcare investments. PMID- 19553765 TI - Costs of end-of-life care: findings from the province of Saskatchewan. AB - As part of the Health Care Use at the End-of-Life in Western Canada Study, the Western Office of the Canadian Institute for Health Information (CIHI) collaborated with the ministries or departments of health in British Columbia, Alberta, Saskatchewan and Manitoba to characterize selected aspects of healthcare at the end of life. In-depth supplementary studies were also conducted for each of the four western provinces. Saskatchewan focused its analysis on healthcare costs in the two years before death. This paper provides a summary of data on healthcare costs for persons who died in Saskatchewan in the 2003-2004 fiscal year. It is based on a more in-depth report prepared for CIHI and Saskatchewan Health titled "Final Report of the Saskatchewan End-of-Life Care Project." PMID- 19553766 TI - Facilitating innovation in the clinical setting: a pathway for operationalizing accountability. AB - Innovation is essential to ongoing improvements in quality healthcare, yet healthcare institutions continue to struggle with the challenge of how best to operationalize the multiple commitments to risk minimization, accountability, the defensible use of resources and quality innovative practices. An ideal pathway would be one that adheres to both the ethical values of the institution in which the innovation is being introduced and relevant legal standards in a way that preserves the innovators' imagination and enthusiasm while ensuring scientific rigour and accountability. An interdisciplinary team at the Hospital for Sick Children in Toronto, Ontario, developed a policy-entrenched pathway f or managing the introduction of innovative procedures in surgery. This article describes the innovation pathway and the steps; taken in its development. PMID- 19553767 TI - Descriptive analysis of the inequalities of health information resources between Alberta's rural and urban health regions. AB - In an effort to understand the extent of the inequalities in health information resources across Alberta, SEARCH Custom, HKN (Health Knowledge Network) and IRREN (Inter-Regional Research and Evaluation Network) conducted a survey in December 2007 to determine what library resources currently existed in Alberta's seven rural health regions and the two urban health regions. Although anecdotal evidence indicated that these gaps existed, the analysis was undertaken to provide empirical evidence of the exact nature of these gaps. The results, coupled with the published literature on the impact, effectiveness and value of information on clinical practice and administrative decisions in healthcare management, will be used to build momentum among relevant stakeholders to support a vision of equitably funded health information for all healthcare practitioners across the province of Alberta. PMID- 19553768 TI - Innovation in managing the referral process at a Canadian pediatric hospital. AB - The provision of timely and optimal patient care is a priority in pediatric academic health science centres. Timely access to care is optimized when there is an efficient and consistent referral system in place. In order to improve the patient referral process and, therefore, access to care, an innovative web-based system was developed and implemented. The Ambulatory Referral Management System enables the electronic routing for submission, review, triage and management of all outpatient referrals. The implementation of this system has provided significant metrics that have informed how processes can be improved to increase access to care. Use of the system has improved efficiency in the referral process and has reduced the work associated with the previous paper-based referral system. It has also enhanced communication between the healthcare provider and the patient and family and has improved the security and confidentiality of patient information management. Referral guidelines embedded within the system have helped to ensure that referrals are more complete and that the patient being referred meets the criteria for assessment and treatment in an ambulatory setting. The system calculates and reports on wait times, as well as other measures. PMID- 19553769 TI - Role of the advanced practice physiotherapist in decreasing surgical wait times. AB - The role of the advanced practice physiotherapist (APP) in outpatient orthopedic clinics has been present in Ontario for over five years. These professionals have additional duties beyond those of a regular physiotherapist in order to screen patients pre- and post-operatively, triage patients for surgery, prescribe conservative management and monitor patients on an ongoing basis. The purpose of this role is to improve patient access to timely surgical care by reducing wait times for hip and knee replacement surgeries. Several positive outcomes have been reported in the literature. It has been established that an APP can effectively manage over 30% of the patients referred to a surgeon for hip or knee replacement surgery because these patients do not require surgery; rather, they require conservative management. PMID- 19553770 TI - Strengthening the value of accreditation: Qmentum - one year later. AB - Accreditation is recognized as one of the most effective ways for health services organizations to systematically examine and improve the quality of their services. Accreditation Canada released the Qmentum accreditation program in February 2008. This article outlines the progress made during the first year of implementation, discusses the current challenges and provides insight into what lies ahead. PMID- 19553771 TI - New accreditation program: university health network's experience with Qmentum. AB - In 2008, University Health Network was surveyed using Accreditation Canada's new Qmentum program. The following article describes UHN's experience rolling out the program to over 12,000 staff, physicians and volunteers. The article also outlines key challenges and lessons learned by the multi-site organization, with a focus on staff engagement, on-site survey preparation and sustainability moving forward. Staff feedback on the Qmentum program was extremely positive, and forecast results from Accreditation Canada were excellent. PMID- 19553772 TI - Tracking emergency department overcrowding in a tertiary care academic institution. AB - Despite the release of a national report describing key markers of emergency department (ED) overcrowding, limited linear data using these markers have been published. We sought to report the degree and trends of ED overcrowding in a typical academic hospital and to highlight some of the key markers of ED patient flow and care. We conducted a prospective study in a large Canadian urban tertiary care teaching hospital that receives approximately 55,000 annual adult ED visits. A database captured demographic and real-time process of care data for each patient from 2000 to 2007. Descriptive data are reported using Canadian Triage and Acuity Scale (CTAS) scores. Over the study period, the ED patient visit volume and presentation times remained predictable. Emergent cases (CTAS levels 1-2) doubled from 8 to 16.6%, and urgent cases (CTAS level 3) increased from 40.2 to 50.3%. Moreover, semi-urgent presentations (CTAS level 4) decreased from 42.4 to 28.8%, and non-urgent cases (CTAS level 5) dropped from 9.4 to 4.3%. The median wait time from triage to bed location increased from two minutes (inter-quartile range [IQR] 1, 46) in 2000 to 27 minutes (IQR 2, 110) in 2007, while the median time from bed location to physician remained constant (29 minutes in 2001 versus 28 minutes in 2007). Overall, admissions increased from 20.4 to 23%. Semi-urgent and non-urgent admissions dropped from 11.5 to 7.4% and 3.2 to 1.8%, respectively. Admitted patients "boarding" in the ED increased from 70,955 hours in 2002 to 118,741 hours in 2007, while the number of emergent and urgent patients leaving without being seen increased by more than 400%. ED overcrowding in a tertiary care hospital is primarily a result of access block due to boarding admitted patients, a situation that poses serious risks to the majority of patients who have emergent or urgent conditions that cannot be managed appropriately in the waiting room. PMID- 19553773 TI - Commentary: Ontario's efforts to reduce time spent in hospital emergency departments. PMID- 19553774 TI - The convenience of flow cytometry for HLA-B*5701 screening to prevent abacavir hypersensitivity reactions. PMID- 19553775 TI - Switch from enfuvirtide to raltegravir in patients with undetectable viral load: efficacy and safety at 24 weeks in a Montreal cohort. PMID- 19553776 TI - Increased frequency of HIV-1 viral load blip rate observed after switching from Roche Cobas Amplicor to Cobas Taqman assay. PMID- 19553777 TI - Gender and care: access to HIV testing, care, and treatment. AB - HIV transmission and occurrence of AIDS in the Middle East and North Africa region (MENA) is increasing, while access to ART in the region lags behind most low to middle-income countries. Like in other parts of the world, there is a growing feminization of the epidemic, and men and women each confront unique barriers to adequate HIV prevention and treatment services, while sharing some common obstacles as well. This paper focuses on important gender dimensions of access to HIV testing, care and treatment in the MENA region, including issues related to stigma, religion and morality, gender power imbalances, work status, and migration. Culturally specific policy and programmatic recommendations for improving HIV prevention and treatment in the MENA region are offered. PMID- 19553778 TI - Gendered empowerment and HIV prevention: policy and programmatic pathways to success in the MENA region. AB - Although HIV in the Middle East and North Africa is currently characterized as a low seroprevalence epidemic, there are numerous factors that are present in the region that could prevent-or exacerbate-the epidemic. The time to invest substantially in prevention-and gender-specific prevention in particular-is now. Given that most policy makers do not make gender-specific plans as epidemics progress, our research team-which draws upon expertise from both within and outside the region-worked together to make programmatic and policy suggestions in the Middle East and North Africa region in 5 key areas: (1) gender-specific and gender transformative HIV prevention interventions; (2) access to quality education and improvements in life skills and sex education; (3) economic empowerment; (4) property rights; and (5) antiviolence. In short, this work builds upon many ongoing efforts in the region and elucidates some of the links between gendered empowerment and health outcomes around the world, particularly HIV and AIDS. PMID- 19553779 TI - Men, HIV/AIDS, and human rights. AB - Though still limited in scale, work with men to achieve gender equality is occurring on every continent and in many countries. A rapidly expanding evidence base demonstrates that rigorously implemented initiatives targeting men can change social practices that affect the health of both sexes, particularly in the context of HIV and AIDS. Too often however, messages only address the harm that regressive masculinity norms cause women, while neglecting the damage done to men by these norms. This article calls for a more inclusive approach which recognizes that men, far from being a monolithic group, have unequal access to health and rights depending on other intersecting forms of discrimination based on race, class, sexuality, disability, nationality, and the like. Messages that target men only as holders of privilege miss men who are disempowered or who themselves challenge rigid gender roles. The article makes recommendations which move beyond treating men simply as "the problem", and instead lays a foundation for engaging men both as agents of change and holders of rights to the ultimate benefit of women and men. Human rights and other policy interventions must avoid regressive stereotyping, and successful local initiatives should be taken to scale nationally and internationally. PMID- 19553781 TI - Gender and HIV in the Middle East and North Africa: lessons for low prevalence scenarios. PMID- 19553782 TI - Twenty-five years of HIV: lessons for low prevalence scenarios. AB - During the initial quarter century since the discovery of HIV, international response has focused on high prevalence scenarios and concentrated epidemics. Until recently, the theoretical underpinnings of HIV prevention were largely based on these responses-the assumption that inadequate responses to concentrated epidemics within low prevalence populations could rapidly lead to generalized epidemics. The limits of these assumptions for HIV prevention in low prevalence scenarios have become evident. While examples of rapid HIV diffusion in once low prevalence scenarios exist, emergence of generalized epidemics are less likely for much of the world. This paper reviews several key issues and advances in biomedical and behavioural HIV prevention to date and highlights relevance to low prevalence scenarios. PMID- 19553783 TI - HIV surveillance and epidemic profile in the Middle East and North Africa. AB - HIV infection is the most devastating infection that has emerged in the recent history. The risk of being infected can be associated with both individual's knowledge and behavior and community vulnerability influenced by cultural norms, laws, politics, and social practices. Despite that the countries in the Middle East and North Africa have succeeded in keeping low the HIV epidemic rates, the number of identified infected cases are increasing. Since the appearance of the first AIDS cases, all the national authorities devoted their efforts to abort the epidemic in its early stages. The rate of new HIV infections across the Middle East and North Africa region are not at an alarming level, but the need for a concerted effort from nation-states and nongovernmental organizations to stem the spread of the virus across the region is vital.Most countries of the region have put in place better information systems to track the HIV epidemic, yet the passive HIV/AIDS reporting remains the cornerstone in the HIV surveillance systems. Several countries still believe that their current strategies are optimal to the HIV status within their territories and that their national strategies are appropriate to their low epidemic status that is not expected to grow. Additionally, these countries fear that establishing an HIV national program to survey risk behaviors may be perceived as an approval of these behaviors that are culturally and religiously unacceptable. This background article aims to summarize the HIV surveillance strategies and epidemic profile in 17 Arab countries in the Middle East and North Africa. The article, also, displays the national surveillance system and the epidemic profile in Egypt and Lebanon as models for the region. This information aims to provide useful insights that may help the national authorities in finding out the best surveillance strategies that allow merging and collecting biological and risk data which is an integral part of their efforts to fight the HIV epidemic in the region. PMID- 19553784 TI - Gender, empowerment, and health: what is it? How does it work? AB - As the HIV/AIDS epidemic has progressed, the role of gender inequality in its transmission has become increasingly apparent. Nearly half of those living with the virus worldwide are women, and women's subordination to men increases their risk of infection and makes it harder for them to access treatment once infected. Men, too, suffer from harmful gender norms-the expectation that they will behave in ways that heighten their risk of HIV infection and that they will be cavalier about seeking health care increases their vulnerability to the disease. In the Middle East and North Africa, HIV infection rates are low, but changing gender norms have the potential to accelerate the spread of the disease if gender inequality is not addressed. Improving women's education, workforce participation, and social and political opportunities is crucial to strengthening health in the region. Work with men to shift gender imbalances is a further important task for the region's policy-makers and civil society groups. PMID- 19553787 TI - Supplement on combination therapy. PMID- 19553788 TI - Combination therapy from the regulatory perspective. AB - For a combination regimen to receive regulatory approval, it must provide some incremental advantage over a previous standard. Although this advantage is typically a greater likelihood of achieving a clinically relevant endpoint, there are others. For example, the additive effects of two agents may allow each to be employed in relatively low doses, improving tolerability. In age-related macular degeneration, one challenge to the combination of pharmaceutical agents is the delivery of adequate quantities of two or more pharmaceutical agents to the back of the eye. The confined space limits accommodation of substantial single dose volumes, whereas repeated invasive delivery through intravitreal injection may be impractical. Innovative delivery strategies may be crucial to future progress. PMID- 19553789 TI - Strategies for inhibiting vascular endothelial growth factor. AB - Clinical trials increasingly support the premise that inhibition of vascular endothelial growth factor is a viable but insufficient target for long-term control of age-related macular degeneration (AMD). Additional biologic therapies targeted at very specific steps in the proliferative signaling pathway are being actively sought as alternatives or adjunctive strategies for the treatment of AMD. This rapidly advancing area of drug development is particularly encouraging because of the growing appreciation for the redundancies and interrelationships between the molecular events. The complexity of this signaling pathway supports the development of combination treatments for optimal control of biologic functions. PMID- 19553790 TI - Combination angiostatic therapies: current status. AB - One of the most important obstacles to combining pharmaceutical agents to treat ocular diseases is the risk of physiochemical reactions. In intraocular administration, these reactions may produce incompatibility, instability, or both. They may change the nature of drug activity, and they may threaten normal cellular function, resulting in lens opacities, corneal toxicity, retinal cell damage, or other adverse outcomes. Although many medications have demonstrated efficacy or have shown promise when administered intravitreally, including antifungals, nonsteroidal antiinflammatory drugs, anti-tumor necrosis factor alpha agents, mammalian target of rapamycin inhibitors, metalloproteinase inhibitors, antiviral agents, antineoplastic compounds, and antivascular endothelial growth factor therapies, these have been typically tested as single agents. The potential for these agents to be combined will be largely determined by their physiochemical compatibility. PMID- 19553791 TI - Age-related macular degeneration. AB - Age-related macular degeneration (AMD) is a significant source of morbidity in aging adults. The damage to the macula and vision loss in advanced forms of wet AMD stems from choroidal neovascularization that originates in the choroid and proliferates through breaks in the Bruch membrane. Although inhibitors of angiogenesis have been a recent advance in the treatment of AMD associated with neovascularization, these agents do not appear to cure the condition. Rather, antiangiogensis agents halt or slow progression of AMD in most cases. The development of additional treatments to provide more effective disease control may depend on addressing several pathophysiologic processes simultaneously. The search for more effective treatments will require a better understanding of the different physiologic processes involved with this disease process. PMID- 19553792 TI - Glucocorticoids and their actions in cells. AB - The biologic responses to the administration of exogenous glucocorticoids (GCs) appear to be specific to cell type. As a result, progress in understanding how to improve the benefit-to-risk ratio of GC therapies for the eye will depend on better characterization of both the receptors and the proteins induced when these receptors are stimulated. The complexity and diversity of the GC receptors in human tissue is underscored by evidence that up to 6,000 genes are expressed or suppressed within hours of GC exposure. The enormous potential to use exogenous GC agents to downregulate processes involved in age-related macular degeneration must be balanced against a similar potential for counterproductive effects. Recent progress predicts an increasing precision with which GC steroids can be used to influence biologic functions. PMID- 19553793 TI - Drug delivery strategies for combination ophthalmic treatments. AB - The most significant obstacle to the combination of drug therapies in age-related macular degeneration (AMD) may be drug delivery. Although the posterior of the eye can be reached by intravitreal injections and implants, AMD is a chronic disease process that is likely to require prolonged or indefinite exposure to antiproliferative agents. In addition to the difficulty of repeat dosing in the vitreal space, ocular anatomy prohibits substantial drug volumes, a limitation further compounded when combining two or more agents. With low penetration to target areas of AMD, pathophysiology, and risk of significant systemic toxicity through systemic drug delivery, innovative strategies for introducing agents to their site of action may be the critical component of improved outcomes. PMID- 19553794 TI - Combination angiostatic therapies: targeting multiple angiogenic pathways. AB - The multiplicity of signaling pathways for new blood vessel formation is suggested by the modest antiangiogenic effects achieved when any single pathway or molecular step is blocked. In an experimental model of neovascularization in newborn mice, highly significant improvements in an antiangiogenic effect are achieved when inhibitors of different pathways of new vessel formation are combined. In this model, neovascularization can be completely inhibited in the majority of animals when at least three pathways are inhibited. When only two pathways are blocked, complete inhibition of neovascularization is less commonly observed but still far more common than when a single pathway is inhibited. For the prevention of new blood vessel formation in neovascular eye diseases like age related macular degeneration, the experimental evidence supports combination therapies that inhibit more than one molecular pathway. PMID- 19553795 TI - General principles of radiation and chemoradiation. AB - High-resolution, three-dimensional functional imaging has improved delivery of radiotherapy to tumors while minimizing damage to adjacent, healthy tissue. A growing array of energy sources and delivery systems, including external delivery of protons and brachytherapy, are also credited with improvement in the therapeutic index of treating diseases with radiation. In many malignancies, the therapeutic effects of radiotherapy can be significantly augmented by coordinated delivery of adjunctive therapies, particularly chemotherapy. In age-related macular degeneration (AMD) and other diseases of the eye, radiotherapy combined with pharmaceutical agents, including antiangiogenesis therapies, have the potential to an additive or synergistic effect. In AMD, the most significant challenge for radiotherapy may be precise delivery to the target and avoidance of damage to surrounding tissues. PMID- 19553797 TI - Overview of radiation trials for age-related macular degeneration. AB - Radiotherapy is a promising adjunctive tool to antiangiogenesis therapies for control of the choroidal neovascularization that characterizes exudative (wet) age-related macular degeneration. Historically, radiation monotherapy sufficient to effectively eradicate choroidal neovascularization has been associated with mixed results; however, newer techniques and delivery platforms have been developed to improve efficacy. The most significant improvements are technical advances that improve the precision of energy delivery, so that tissue destruction remains confined to the target. In addition, several combination therapies are showing promise for enhanced effect. Other strategies, such as pretreating neovascular tissue to increase its sensitivity to radiation, thereby reducing the energy dose, may also be viable. However, even though the modern delivery systems permit relatively low dosages, there are risks of radiotherapy to ocular tissue, and its role remains speculative, pending results of ongoing trials. PMID- 19553796 TI - Modulating the tumor microenvironment to improve radiotherapy. AB - Radiotherapy can ablate vascular tissue, but the antiangiogenic effect is counteracted by upregulation of endogenous proangiogenic signals, particularly vascular endothelial growth factor. This effect, which has long been a concern in oncology, is relevant to a discussion of radiation for the treatment of age related macular degeneration (AMD). In oncology, a broad array of agents that target growth factors and receptors is being actively studied for the potential to enhance the activity of radiation in controlling malignancies. Progress with such combinations in oncology may be relevant to parallel strategies for treating and containing neovascularization in AMD and other retinal diseases. PMID- 19553798 TI - Approaches toward combining photodynamic therapy with pharmaceuticals that alter vascular microenvironment. AB - Photodynamic therapy has been widely replaced by antiangiogenesis agents for the first-line therapy for exudative age-related macular degeneration (AMD). There is a strong basis for predicting that a combination of photodynamic therapy and antiangiogenesis agents may address the relative disadvantages of each. By improving the rates of response, photodynamic therapy has the potential to reduce the frequency with which intravitreal injections of antiangiogenesis agents are required. Antiangiogenesis agents may augment the activity of photodynamic therapy by inhibiting its counterproductive upregulation of vascular endothelial growth factor. Clinical studies of this combination are being advanced in both AMD and in the treatment of malignancies. PMID- 19553799 TI - Pathology beyond neovascularization: new targets in age-related macular degeneration. AB - Control of choroid neovascularization may be important but insufficient to preserve long-term vision threatened by age-related macular degeneration. A retrospective analysis of patients who were early participants in anti-vascular endothelial growth factor studies, other pathologic processes, particularly fibrosis and atrophy, have participated in vision loss independent of new vessel growth. The recent interest in combination treatment strategies has been dominated by more effective blockade of angiogenic signaling, but it may also be necessary to incorporate therapies that block fibrosis, inhibit atrophy or other pathophysiologic processes not directly related to neovascularization to achieve the ultimate goal of preserving sight for a long term. PMID- 19553800 TI - The rationale for drug combinations in age-related macular degeneration. AB - The strategies for improving control of age-related macular degeneration with combination therapies are evolving. The focus on more effective blockade of choroidal neovascularization has shifted to include control of additional processes implicated in disease progression, such as neural death, inflammation, or fibrosis. Although there is likely to be a strong interrelationship between choroidal neovascularization and inflammation that contributes to advanced stages of disease progression, including fibrosis, combining treatment strategies may enlarge the opportunity to prevent both early and late vision loss. PMID- 19553801 TI - Combination therapy for age-related macular degeneration. AB - Vascular endothelial growth factor (VEGF) is an important mediator of angiogenesis in age-related macular degeneration (AMD) and is a validated therapeutic target. However, combination of anti-VEGF agents with complementary inhibition of other mediators of angiogenesis, such as platelet-derived growth factor and integrin alpha5beta1, may result in enhanced visual acuity. Other concomitant treatments, such as inhibitors of inflammation, may generate an even stronger barrier to the progression of AMD than that now observed in individuals receiving anti-VEGF therapies alone. Experimental studies are providing support for the general principles of combination treatment in AMD. PMID- 19553802 TI - Combination therapy using the small interfering RNA bevasiranib. AB - Bevasiranib, the first small interfering RNA agent developed for the treatment of neovascular age-related macular degeneration, has demonstrated clinical promise. Injected intravitreally, this small interfering RNA acts by inducing catalytic destruction of messenger RNA to silence gene expression. Bevasiranib targets the production of vascular endothelial growth factor (VEGF) protein. It does not affect existing VEGF protein, suggesting that it may offer a synergistic effect when given in combination with anti-VEGF treatments, such as ranibizumab. The safety of bevasiranib has been supported by preclinical and clinical research. PMID- 19553803 TI - Rationale for combination therapy in age-related macular degeneration. AB - Choroidal neovascularization (CNV) is typically characterized as an invasion of blood vessels, but it is important to recognize concurrent inflammatory and mesenchymal cell infiltration. A two component model of CNV can be thought to be composed of a vascular component and an extravascular component. Macular damage is possible through either. Given the redundancy and complex interaction among biologic systems involved in the production of CNV, it is likely that a monotherapeutic approach will not be fully effective. The vascular component of CNV arises through the orchestrated interaction of a number of growth factors such as vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF), which maintains pericyte viability. The vascular component of CNV can be selectively targeted with anti-VEGF and anti-PDGF drugs or nonselectively with such modalities as ionizing radiation. The extravascular component can be targeted selectively by inhibiting specific cytokines such as tumor necrosis factor alpha or nonselectively with antiinflammatory drugs such as corticosteroids. PMID- 19553804 TI - Opportunities and challenges in the development of combination therapy for the treatment of retinal diseases. AB - Ranibizumab, a humanized monoclonal antibody fragment (fragment, antigen binding, FAB) that neutralizes all of the soluble isoforms of vascular endothelial growth factor (VEGF)-A, was a significant step forward in the control of age-related macular degeneration. However, this agent, like others with the same mechanism, has several limitations. Although ranibizumab preserves vision in almost all patients, only a fraction achieves a halving of the visual angle. By inhibiting the activities of VEGF, ranibizumab blocks the continued growth of choroidal neovascularization, but existing neovascular lesions do not regress. Further, ranibizumab addresses only the increased production of VEGF; it does not address the underlying cause of enhanced cytokine production. Although ranibizumab is well tolerated, the course of the disease is unpredictable, necessitating frequent patient monitoring and treatment. Although strategies that guide retreatment based on the reoccurrence of retinal edema can be used to reduce treatment burden, this strategy may not provide optimal patient benefit. Because of these limitations, the identification of effective adjunctive therapies is considered an urgent goal. PMID- 19553805 TI - Triple therapy for age-related macular degeneration. AB - Choroidal neovascularization is a hallmark sign of wet age-related macular degeneration (AMD) but it is not an isolated feature. Several processes are likely to contribute to the fibrotic scarring and vision loss that accompanies progressive disease. In a case series, a triple therapy approach to wet AMD was based on the goals of halting choroidal neovascularization, controlling the inflammatory response, and modifying proliferative factors. To address each of these goals, respectively, patients received photodynamic therapy, bevacizumab, and the steroid dexamethasone. The encouraging rate of response, including significant improvements in visual acuity, is consistent with the combined activities of these agents and provides the basis for more definitive studies. PMID- 19553806 TI - Elevated elafin/trappin-2 in the female genital tract is associated with protection against HIV acquisition. AB - OBJECTIVES: Globally, heterosexual intercourse is the primary route of HIV-1 (HIV) transmission. It follows that mechanisms that protect against HIV infection are likely operative at the genital mucosa. In HIV-resistant Kenyan sex workers who are highly exposed to HIV infection yet remain uninfected, protection correlates with HIV-specific immune responses and genetic factors. However, these factors do not entirely explain this model of natural immunity to HIV. We hypothesized that protection may be mediated by innate immune proteins in the genital tract of HIV-resistant sex workers. DESIGN AND METHODS: The genital proteome of mucosal secretions from HIV-resistant women was examined using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Cervical lavage samples were collected from 315 HIV-resistant, HIV-uninfected and HIV-infected commercial sex workers. RESULTS: Univariate analysis identified a 6 kDa biomarker of HIV resistance in genital secretions from these women. This protein was identified by tandem mass spectrometry as elafin and was found to be overexpressed in HIV-resistant women compared with HIV-uninfected (P = 0.001) and infected (P = 0.002) women. The elevated levels of elafin/trappin-2 in HIV resistant women were confirmed using ELISA. The prospective association of elevated cervicovaginal elafin/trappin-2 levels with protection from HIV acquisition was then confirmed in an independent cohort of high-risk female sex workers. CONCLUSION: Using a unique proteomics approach in a large scale, cross sectional cohort study, we identified elafin/trappin-2 as a novel innate immune factor, which is highly associated with resistance. This association was confirmed within an independent, prospective cohort study. Genital tract elafin/trappin-2 levels constitute a natural correlate of HIV protection in humans. PMID- 19553808 TI - Healthcare-associated infections. A useful concept? AB - PURPOSE OF REVIEW: To provide an overview of the importance of healthcare associated infections (HCAI) as a new concept in infectious diseases. Although described originally in bacteremia, the concept has also been applied to other infections such as pneumonia and endocarditis. RECENT FINDINGS: Therapeutic protocols recommend treating HCAI with broad-spectrum antibiotics to cover infection due to multidrug-resistant pathogens (MDR). Nonetheless, the prevalence of MDR pathogens may vary considerably in different subgroups of HCAI and in different countries. Patients categorized as having HCAI usually present atypical symptoms that may delay diagnosis; moreover, outcome is worse than community acquired infections. Although several studies have reported that inappropriate empiric therapy may explain adverse outcomes in HCAI, careful adjustment for other conditions should be taken into account, such as baseline comorbidities, therapeutic limitations, and delay in the initiation of antibiotic treatment. SUMMARY: In patients with HCAI, complementary workup should be systematically performed in search of a microbiologic diagnosis. Broad-spectrum antibiotics should not be prescribed automatically, especially in countries with lower prevalence of MDR pathogens. Some refinements of the definition are needed and specific risk factors for infection by MDR microorganisms should be assessed. PMID- 19553807 TI - Association of hepatitis C virus and HIV infection with subclinical atherosclerosis in the women's interagency HIV study. AB - Whether hepatitis C virus coinfection might accelerate atherosclerosis in HIV infected individuals is unclear. We examined the relationship of HIV and hepatitis C virus with carotid artery intima media thickness and the presence of carotid plaques in the Women's Interagency HIV Study. Hepatitis C virus infection was not associated with greater carotid artery intima media thickness after adjustment for demographic and traditional cardiovascular risk factors. Further follow-up is needed to clarify whether HIV/hepatitis C virus coinfection may be associated with a greater risk of carotid plaque. PMID- 19553809 TI - Cardiovascular problems in noncardiac surgery. AB - PURPOSE OF REVIEW: Perioperative cardiac complications remain a major area of concern as our surgical population increases in volume, age and frequency of comorbidity. A variety of strategies can be used to optimize patients and potentially reduce the incidence of these serious complications. RECENT FINDINGS: Recent literature suggests a trend towards less invasive testing for detection and quantification of coronary artery disease and greater interest in pharmacologic 'cardioprotection' using beta-blockers, statins and other agents targeting heart rate control and other mechanisms (e.g. reducing inflammatory responses). The recent Perioperative Ischemic Evaluation study has substantially altered this approach at least towards widespread application to lower/intermediate risk cohorts. Considerable attention has been focused on ensuring optimal standardized perioperative management of patients with a recent percutaneous coronary intervention, particularly those with an intracoronary stent. Widespread surveillance of postoperative troponin release and increasing recognition of the prognostic potential of elevated preoperative brain natriuretic peptides point towards changing strategies for long-term risk stratification. SUMMARY: The complexity of a particular patient's physiologic responses to a wide variety of surgical procedures, which are undergoing constant technological refinement generally associated with lesser degrees of invasivity and stress make calculation of patients' perioperative risk very challenging. At the present time, adequate information is available for the clinician to screen patients with high-risk preoperative predictors, delay elective surgery for patients with recent intracoronary stents and continue chronic beta-blockade in appropriate patients. New large-scale database and subanalyses of major trials (e.g. Perioperative Ischemic Evaluation and Coronary Artery Revascularization Prophylaxis) should provide additional information to minimize perioperative cardiac risk. PMID- 19553811 TI - Thrombopoietin receptor agonists in the treatment of thrombocytopenia. AB - PURPOSE OF REVIEW: The basis of treatment for immune thrombocytopenic purpura (ITP) has conventionally relied on nonspecific immune suppression designed to reduce platelet destruction. As a consequence, at least half of the morbidity and mortality in this condition is related to infection secondary to treatment, and alternative treatments are desirable. RECENT FINDINGS: It has been shown that ITP is not purely due to platelet destruction, and in a significant proportion platelet production is suboptimal. Further interest developed with the discovery that the recombinant thrombopoietins (TPOs) could enhance platelet production in a variety of thrombocytopenic states. With the development of the second generation of TPOs, which had no sequence homology to endogenous TPOs, studies confirmed clinical effect. Two agents, romiplostim and eltrombopag, are now licensed and their place in the treatment is being evaluated. SUMMARY: Platelet responses to romiplostim and eltrombopag are seen in a much greater percentage than in other second-line studies, and these are maintained while the drugs continue to be administered. Both are well tolerated, with no significant adverse effects over placebo, and have an effect both presplenectomy or postsplenectomy. An interesting initial observation has been that the platelet response is associated with an improved quality of life in many patients when compared with conventional management. PMID- 19553810 TI - Kindling the flame of integrin activation and function with kindlins. AB - PURPOSE OF REVIEW: There are three kindlin family members in vertebrates, which have high-sequence homology and a common organization signature with a C-terminal 4.1, ezrin, radixin, moesin (FERM) domain bisected by a pleckstrin-homology domain. Although the cell and tissue distributions of the three kindlins differ, there is a consistent and close interrelationship between kindlins and integrins, and alterations of kindlin expression affect integrin-dependent functions. However, in-vivo data on the functions of the kindlins and their mechanisms of action have been lacking. RECENT FINDINGS: Recent studies have shown that deficiencies of each of the three kindlins result in profound and distinct phenotypes, ranging from skin and intestinal defects (kindlin-1), embryonic lethality due to cardiac developmental problems (kindlin-2), to marked abnormalities in platelet, leukocyte and erythrocyte function (kindlin-3). A human disease characterized by bleeding, frequent infections and osteopetrosis has now been attributed to mutations in the gene encoding for kindlin-3. These defects are consistent with recent data showing that kindlins control integrin activation and function. SUMMARY: The three members of the kindlin family have now been implicated as essential regulators of integrin function in individual cells and in whole organisms. PMID- 19553812 TI - Muscle atrophy in immobilization and senescence in humans. AB - PURPOSE OF REVIEW: Recent reports exploring the mechanisms thought to be responsible for the determination of muscle mass during health, ageing and immobilization in humans have presented findings that have wide ranging implications. This brief review highlights some of the more important findings. RECENT FINDINGS: Contrary to expectations, recent findings suggest an apparent dissociation between muscle signalling pathways and their associated events in humans, particularly in relation to muscle protein synthesis. Although debate concerning the relative importance of muscle protein synthesis and degradation to muscle mass loss during immobilization continues, and the mechanisms responsible for this loss and its restoration during rehabilitation remain unclear, new evidence has emerged showing that anabolic resistance to protein nutrition develops during immobilization. This latter observation is in agreement with earlier evidence pointing to anabolic resistance of muscle to protein nutrition existing in the elderly, which is of clinical importance. Recent observations also suggest that a sex difference exists in the rate of muscle protein synthesis under postabsorptive conditions in the elderly and may explain why women lose muscle mass at a slower rate with age than men. SUMMARY: These recent findings highlight our current lack of understanding of the mechanisms that regulate muscle mass in humans. PMID- 19553813 TI - Tremor. AB - PURPOSE OF REVIEW: Tremor continuously attracts the attention of clinicians and basic researchers in search of pathophysiological, molecular and genetic mechanisms of the oscillatory activity. RECENT FINDINGS: A widespread dynamic network of cortical and subcortical oscillators taking part in tremor generation intermittently has been postulated. Essential tremor is accompanied by functional deficits but may also occur along with subtle cerebellar changes. According to recent epidemiological studies there may be a link of essential tremor with Parkinson's disease. Many of the epidemiologic studies suffer from small cohorts, small effects or the lack of a definite test for essential tremor leaving the diagnosis a pure clinical one. A very recent large genome-wide association study has revealed that the LINGO1 gene is associated with an increased risk for essential tremor. Topiramate is becoming the best-established second line treatment for essential tremor. Targets for deep brain stimulation in the grey matter below the ventral intermediate nucleus of the thalamus seem to be most effective. SUMMARY: New concepts of the central origin of tremors stimulate the search for new therapeutic targets for tremor suppression outside the basal ganglia and thalamus (e.g. cortex). The role of structural neurodegenerative changes in essential tremor remains an open question. Further studies on specific subgroups of patients are necessary. PMID- 19553814 TI - Role of PET in gynecologic malignancy. AB - PURPOSE OF REVIEW: The purpose of this review is to outline the current state-of the-art use of PET imaging for patients with gynecologic malignancies. RECENT FINDINGS: Recent findings with clinical impact when PET is utilized in patients with gynecologic cancer are primarily limited to patients with cancer of the uterine cervix and for patients with ovarian cancer. PET is used in patients with other gynecologic cancers such as endometrial cancer, uterine sarcomas, vulvar cancer, and vaginal carcinoma but with less well defined clinical impact than cervical and ovarian cancers. SUMMARY: PET is utilized in patients with cervical cancer for initial staging, guiding therapy, evaluating response to therapy, and for long-term follow-up. The primary uses of PET in patients with ovarian cancer are for the evaluation of adnexal masses and for the diagnosis and evaluation of recurrent disease. PMID- 19553815 TI - Promising molecular targets in ovarian cancer. AB - PURPOSE OF REVIEW: Clinically and on a molecular level, ovarian cancer is a unique and complex disease. The explosion in potential molecular targets over the last decade has led to the arrival of many novel therapies into oncology. In the present article, we review the most promising of these agents in ovarian cancer. RECENT FINDINGS: Targeted therapies, such as epidermal growth factor receptor inhibitors, that have worked well in other cancers have shown only moderate success in ovarian cancer, whereas other treatment approaches have yielded surprisingly positive outcomes. An example is anti-vascular endothelial growth factor and proapoptotic strategies, which are effective in both primary and relapsed ovarian cancer. Use of poly (ADP-ribose)-polymerase inhibitors has shown that targeting one form of DNA repair profoundly affects cell survival in those with a hereditary failure to mend DNA damage using another mechanism. This can be extrapolated to patients with sporadic ovarian cancers, with or without the 'BRCAness' phenotype. SUMMARY: Using targeted agents in ovarian cancer, we are discovering not only how these novel therapies work but are also unveiling the complex 'wiring' of the disease itself, and the interconnections between what were previously believed to be distinct molecular pathways. The addition of targeted agents to our therapeutic armoury is likely to significantly and positively impact on patient survival. PMID- 19553816 TI - Vertebral augmentation in osteoporotic and osteolytic fractures. AB - PURPOSE OF REVIEW: To discuss recent advances in percutaneous minimally invasive vertebral augmentation techniques used in the treatment of osteolytic and osteoporotic compression fractures. RECENT FINDINGS: Recent data have shown that outcomes in patients with vertebral compression fractures treated with vertebral augmentation techniques have been encouraging. Several studies demonstrated the positive effects of vertebral augmentation on maintenance of postoperative pain reduction at long-term follow-up. There is more evidence on the relationship between vertebral augmentation and adjacent segment vertebral compression fractures. In addition, the literature verifies that good outcomes can be expected in patients with spinal metastases and multiple myeloma. SUMMARY: The incidence of symptomatic vertebral compression fractures is on the rise. For those patients with severe pain or progressive collapse due to osteoporotic or osteolytic vertebral compression fractures, early vertebral augmentation affords excellent early pain relief, early return to function, and restores and maintains sagittal alignment. PMID- 19553817 TI - Predicting life expectancy in prostate cancer patients. AB - PURPOSE OF REVIEW: Due to its long natural history, prostate cancer illustrates best the need for tools that adequately predict life expectancy. We reviewed the actual tools available for clinicians involved in therapeutic decisions in newly diagnosed prostate cancer and examined their accuracy to provide individual life expectancy. RECENT FINDINGS: Life tables, comorbidity indices, and multivariate prognostic models can assist clinicians for life expectancy predictions. However, the accuracy of life tables (60.9%) and comorbidity indices (accuracy unknown) may be as weak as clinician-derived life expectancy predictions (69%). Actually, statistical models provide the highest accuracy (69-84.3%). To date, Walz et al. developed the most accurate model (84.3%), predicting the risk of death of nonprostate cancer-related causes within 10 years of definitive therapy. SUMMARY: Clinicians need the most accurate estimates of life expectancy in situations in which there is uncertainty regarding the need for aggressive local therapy. As the accuracy of clinician-derived life expectancy prediction is relatively modest, clinicians may benefit from assisted life expectancy prediction by life tables and statistical tools in their daily clinical practice. This would enhance the accuracy of the life expectancy predictions of individual candidates to definitive therapy for prostate cancer. Actually, nomograms provide the most accurate health-adjusted life expectancy prognostication. PMID- 19553818 TI - The current management of small renal masses. AB - PURPOSE OF REVIEW: Small renal masses are increasingly being discovered incidentally on routine abdominal imaging performed for other purposes. The natural history of small renal masses as well as the therapeutic approach for such lesions is an evolving paradigm. In this review, we assess the current literature regarding this controversial topic. RECENT FINDINGS: Nephron-sparing surgery can be performed with minimal morbidity and mortality. Ablative techniques, such as cryoablation and radio frequency ablation, have shown both safety and reliability in series reporting short-term oncologic follow-up. Preliminary studies seem to favor cryoablation when compared with radio frequency ablation. Observational series have shown that very few of these selected patients present disease progression while on active surveillance. Finally, recent studies have shown that renal biopsy is both well tolerated and accurate. SUMMARY: The management of small renal masses remains a challenging issue. Although nephron-sparing surgery is the standard of care for these masses, ablative techniques and observation represent adequate alternatives, especially for poor surgical candidates. Renal biopsy is increasingly proving to be of value in the management of small renal masses. PMID- 19553819 TI - Predicting favourable prognosis of urothelial carcinoma: gene expression and genome profiling. AB - PURPOSE OF REVIEW: During the past few years, information on (epi)genetic and expression profiling of urothelial carcinomas has expanded, allowing a better appreciation of their correlation with clinicopathological features of bladder cancer. RECENT FINDINGS: The two-pathway model of bladder carcinogenesis separating a favourable pathway characterized by mutations in the fibroblast growth factor 3 gene (FGFR3) and a clinically unfavourable pathway characterized by genetic instability and mutations in the p53 gene is now well established. Noninvasive (pTa), superficially invasive (pT1) and muscle invasive (pT2) bladder cancers can be separated statistically on the basis of extent of genomic instability. Expression (cDNA) array analyses are able to define mRNA signatures specifically associated with the two pathways of bladder carcinogenesis. Currently, attention is shifting to the role of epigenetic alterations in bladder carcinogenesis, including promoter hypermethylation of specific genes and aberrant expression of microRNAs. The level of promoter hypermethylation gradually increases from morphologically normal urothelium to invasive carcinoma. Aberrant expression of specific microRNAs is specifically related to the FGFR3 mutant defined bladder carcinogenesis pathway. SUMMARY: Quantitative genomic (DNA) alterations are associated with the two major molecular pathways of bladder carcinogenesis, defined by FGFR3 and p53 mutations. Chromosomal alterations, cancer specific mRNA expression signature and promoter hypermethylation may precede clinically and histopathologically detectable bladder cancer. As gene expression signature, promoter hypermethylation of selected genes and aberrant expression of some microRNAs are promising as bladder cancer biomarkers, future studies should explore their potential clinical significance taking into account their robustness and cost-effectiveness. PMID- 19553820 TI - Environmental factors promoting bladder cancer. AB - PURPOSE OF REVIEW: To understand the molecular mechanisms of bladder carcinogenesis in relation to environmental carcinogens in order to provide a given population with a preventive value of bladder cancer. RECENT FINDINGS: Cigarette smoking, aromatic amines contained in dyes, chronic inflammation due to infection such as schistosomiasis, anticancer drugs, drug abuse of analgesic, and radiation are considered as well known risk factors of bladder cancer. Several environmental factors are supposed to be involved in carcinogenesis, cancer progression, and patient's prognosis in bladder cancer. On the basis of the results of recent genetic studies in relation to bladder carcinogenesis, several genetic polymorphisms of detoxification or DNA repair such as N-acetyltransferase 2, glutathione S-transferases, and human 8-oxoguanine DNA glycosylase 1 give us important information in relation to environmental risk factors and ethnic differences for predicting the prognosis of patients with bladder cancer. SUMMARY: Prevention of environmental carcinogens is important from the viewpoint of the social and clinical problems since elucidation of the correlation between epidemiologic and genetic phenomenon enable us to improve the life expectancy and quality of life of bladder cancer patients. PMID- 19553821 TI - Perioperative instillation therapy in superficial bladder cancer: is it effective regarding outcome and costs? AB - PURPOSE OF REVIEW: This article reviews the current knowledge concerning immediate intravesical instillation therapy in nonmuscle-invasive bladder cancer after transurethral resection of the bladder tumour, with emphasis on the literature of the last few years. RECENT FINDINGS: A review was conducted on the recent literature available by PubMed database on the subject of immediate bladder chemotherapy after transurethral resection of the bladder tumour and its recent developments. SUMMARY: A single immediate bladder instillation with chemotherapy will give 39% reduction of recurrence. Numbers needed to treat to prevent one recurrence are estimated at 8.5. In intermediate and high-risk bladder cancer, the immediate postoperative instillation does not give sufficient reduction in recurrence rate to leave out subsequent bladder instillations. The significant reduction of recurrences, the mild side-effects of the treatment and the clear cost-effectiveness make one immediate instillation a valuable addition to transurethral resection of the bladder tumour in the treatment of nonmuscle invasive bladder cancer. Points of controversy are the true numbers needed to treat to prevent a recurrence and the type of recurrences that will be prevented. No new types of instillation are added recently to the ones available, but promising types are under investigation of which pharmacokinetic studies have shown acceptable rates of side-effects. PMID- 19553822 TI - The risk and prophylactic management of bladder cancer after various forms of radiotherapy. AB - PURPOSE OF REVIEW: Prostate cancer is the most common cancer diagnosed in men and remains the second most lethal malignancy. Most patients undergoing treatment elect for radical prostatectomy or radiation. As the number of patients treated has increased and survival improved, delayed complications of these modalities has assumed increased importance. Recent studies report an increased risk of certain cancers after radiation for prostate cancer. This review aims to summarize recent data. RECENT FINDINGS: Recent studies have confirmed the association of prostate radiation with secondary cancers. The most common secondary malignancy is bladder carcinoma. We have treated 44 patients with bladder cancer who had radiation therapy for prostate cancer. At diagnosis, 60% had tumor, which invaded the bladder muscle (T2 or greater disease). The mean latency from radiation to diagnosis of bladder cancer was 5.5 years. SUMMARY: Radiation therapy for prostate cancer is associated with an increased risk of bladder cancer. In our series, patients presented at higher stage than expected from population-based studies of bladder cancer. Patients and their physicians should be aware of such risks when choosing therapy for prostate cancer. Hematuria following radiation therapy for prostate cancer should be investigated rather than being attributed to radiation-induced cystitis. PMID- 19553823 TI - Does extended lymphadenectomy preclude laparoscopic or robot-assisted radical cystectomy in advanced bladder cancer? AB - PURPOSE OF REVIEW: Open radical cystectomy with an appropriate bilateral lymph node dissection (LND) is currently the standard treatment for patients with muscle-invasive bladder cancer. Approximately 25% of patients with stages T1-T4 N0 M0 harbour metastatic lymph nodes at the time of radical cystectomy. Results from open high volume radical cystectomy series suggest that a more extended LND provides the best survival outcomes and the lowest local recurrence rates. Currently, there is controversy whether laparoscopic or robot-assisted extended LND at radical cystectomy is technically feasible and whether it can provide oncological control equivalent to open LND series at the time of radical cystectomy. RECENT FINDINGS: Laparoscopic LND is technically demanding and requires prolonged operation time. Most studies to date indicate that fewer nodes are removed than with an open approach, putting a question mark to this surgical approach from an oncological point of view. Limited data on lymph node yield using a robot-assisted approach are available; however, several series found similar results as in open series. SUMMARY: At present, there is no conclusive evidence showing that laparoscopic LND gives similar results than open LND. Robot assisted LND is still in its learning curve and more patient series are needed. PMID- 19553824 TI - Genome-wide association studies in bladder cancer: first results and potential relevance. AB - PURPOSE OF REVIEW: The role of genetic susceptibility in the development of urinary bladder cancer is unclear, as it is in many other types of cancer. Since 2007, however, an innovative research approach (i.e. genome-wide association studies or GWASs) has led to the identification of numerous genomic loci that harbor susceptibility factors for one or more cancer sites. All GWASs have been published in high-impact journals and the strengths of the design are acknowledged by all experts, but there is criticism about the relevance of the results. Late 2008, the first GWAS in bladder cancer was published. RECENT FINDINGS: In this review, the principles of GWASs are explained, as well as their strengths and limitations. The study in bladder cancer among 4000 cases and 38,000 controls identified three new susceptibility loci at 8q24, 3q28, and 5p15 that increase the risk of bladder cancer by 22, 19, and 16%, respectively. The results of two other GWASs in bladder cancer are expected to appear this year. Joint analysis of the three studies will probably identify additional susceptibility loci. SUMMARY: The results of bladder cancer GWASs may point the way to yet unknown disease mechanisms. So far, the findings are not sufficiently discriminative for risk predictions to be used in clinical care or public health. PMID- 19553825 TI - Surprisingly high prevalence of subtype C and specific HIV-1 subtype/CRF distribution in men having sex with men in Senegal. AB - BACKGROUND: Recent reports showed the high vulnerability for HIV infection of men who have sex with men (MSM) in Africa. Here, we report the HIV-1 variants that circulate among MSM in Senegal. METHODS: HIV-1 subtype/circulating recombinant form (CRF) was determined in an 1800-base pair fragment of pol for 70 HIV-1 positive samples from MSM. Phylogenetic trees were constructed using the neighbor joining method with CLUSTALX. Similarity and bootstrap plots were then done for recombination analysis. The maximum likelihood approach was used for the identification of transmission clusters. RESULTS: Sixty-seven samples (95%) were from Senegalese MSM, 90% unmarried with a median age of 30 years. Fifty-five MSM had regular male partners, but 39 of 70 had also a regular female partner. The overall subtype/CRF distribution was as follows: 28 C (40%), 17 CRF02_AG (24.3%), 13 B (18.6%), 6 G (8.6%), 3 CRF09_cpx (4.3%), and 3 (4.3%) unique recombinants. In addition, 47 sequences (67.15%) were segregated into 15 transmission clusters. CONCLUSIONS: These variants circulate also among the general population or female sex workers, but the proportions are significantly different. Despite the massive stigma, the majority (80%) of MSM recognized having sex with women and could serve as a bridge for intermixing of HIV-1 variants between high-risk men and low risk women. PMID- 19553827 TI - What can ongoing clinical trials of anticoagulants demonstrate? AB - As the population ages, the burden of thromboembolic disease increases. The development of new anticoagulants that overcome the shortcomings of the vitamin K antagonists represents an important advance.Clinical evaluation of new anticoagulants typically begins in short-term indications, such as prophylaxis against venous thromboembolism in patients undergoing orthopaedic surgery, followed by investigation in chronic conditions, such as stroke prevention in patients with atrial fibrillation. Factors for consideration in clinical trials include methodological issues (blinded versus open-label; active-control statistical designs; patient selection, etc.). Despite its subsequent withdrawal, clinical trials of ximelagatran demonstrated the efficacy of fixed doses of an oral, direct thrombin inhibitor for prevention of thromboembolism. Direct and indirect inhibition of activated Factor X is another target for new anticoagulants. Evidence-based data from trials such as these will pave the way for new anticoagulants, with the goal of bringing optimum prophylactic therapy to those requiring anticoagulation. PMID- 19553826 TI - Is 1 alanine transaminase >200 IU enough to define an alanine transaminase flare in HIV-infected populations? A new definition derived from a large cohort study. AB - OBJECTIVES: Recent studies have suggested that highly active antiretroviral therapy may lead to rises in alanine transaminase (ALT) among HIV-infected patients. However, the definition of an ALT flare is arbitrary and the extent to which such increases represent normal fluctuations has not been explored. METHODS: Using data from untreated, hepatitis B virus/hepatitis C virus-negative, HIV-infected patients, we derived a definition for an ALT flare by exploring a series of ALT thresholds (from 100 to 200 IU/L). The resulting definition (2 consecutive ALTs > 200 measured >2 weeks apart) was applied to all patients in the UK Collaborative HIV Cohort (CHIC) Study, and Poisson regression was used to identify factors associated with ALT flares. RESULTS: Five hundred and twenty six of 12,206 eligible patients (4.3%) had > or =1 ALT flare, resulting in a total of 615 episodes of ALT flares. The overall rate of an ALT flare was 1.19 (95% confidence interval: 1.10 to 1.28) per 100 person-years. Higher risk of ALT flare was associated with lower CD4 counts, detectable viral loads, being under follow up in earlier calendar years, prior clinical AIDS, receipt of nevirapine either with didanosine/stavudine or without didanosine/stavudine, receipt of ritonavir, detectable anti-hepatitis C virus, and detectable hepatitis B surface antigen. CONCLUSIONS: Associations between known risk factors may be under/over estimated if using single values, that is, 1 ALT > 200, to define ALT flares. We recommend studies to use a more stringent measure and suggest our derived definition of an ALT flare. PMID- 19553828 TI - Interleukin-6 and tumor necrosis factor-alpha as biochemical markers of heart failure: a head-to-head clinical comparison with B-type natriuretic peptide. AB - BACKGROUND: The reliability of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) as biochemical markers of heart failure in comparison to B-type natriuretic peptide (BNP) has not been investigated in depth. AIM: To compare the correlations between IL-6, TNF, BNP plasma concentrations and some clinical and instrumental variables and their prognostic value in heart failure patients. METHODS: In 79 patients with heart failure, the correlations between IL-6, TNF and BNP plasma concentrations and a series of 18 variables were studied. Outcome events were death from any cause and combined death and heart transplantation. RESULTS: At univariate analysis, BNP and IL-6 plasma concentrations correlated with each other (r = 0.4828; P < 0.0001), with New York Heart Association class, fluid retention, left ventricular ejection fraction, mean right atrial pressure, mean pulmonary pressure and cardiac index. All these correlations were stronger with BNP. TNF plasma concentration correlated only with New York Heart Association class and left ventricular ejection fraction.During follow-up, 1-32 months, 14 patients died and nine underwent heart transplantation. At univariate analysis, both BNP and IL-6 plasma concentrations were predictors of death and heart transplantation, but only BNP was a predictor of death; however, only creatinine plasma level was an independent predictor of prognosis. CONCLUSION: IL 6 and TNF are less reliable biochemical markers than BNP in heart failure patients. PMID- 19553829 TI - Congenital anomalies of the hand: an overview. AB - Of the 1% to 2% of newborns that are born with congenital defects, 10% of these are born with upper extremity malformations. Although many classification systems have been developed, the most widely used classification system was developed by Swanson. This system categorizes the congenital upper extremity malformations according to the embryonic process that has failed to develop. Congenital hand malformation is a broad category that is broken down into 7 subclasses. This review will focus on congenital hand defects and their associated craniofacial syndromes. The topics will include failure of formation, failure of differentiation, duplication, overgrowth, undergrowth, constriction band syndromes, and generalized skeletal abnormalities. PMID- 19553830 TI - Replantation in the pediatric hand. AB - Replantation in the pediatric hand is not an uncommon procedure, and it is 1 of the most common microsurgical operations in children. Microsurgeons should be aware of the indications and technical issues involving pediatric patients because there are differences compared to adult patients. In general, pediatric patients can be approached in a liberal and aggressive manner, but they tend to be more technically challenging. This review attempts to describe the various considerations in dealing with these patients to provide a framework for surgical management. Based on a survey of the literature, replantation has been a successful procedure in the pediatric population and has become a mainstay of treatment because it provides a better functional and aesthetic outcome. PMID- 19553831 TI - A new consideration of scar formation in open rhinoplasty. AB - Rhinoplasty is one of the most variable operations in facial plastic surgery because of the various deformities involved and dissimilar methods used to correct them. Although the exposure of the surgical anatomy with an open approach to rhinoplasty is much better than with the endonasal approach, many surgeons focus mainly on the risk of a visible columellar scar. However, despite this unfavorable complication of a midcolumellar incision, a better understanding of the nasal deformities can be gained, and a more detailed reconstruction can be performed. Surgeons have developed various methods to reduce the risk of scar formation. Different incision types include the Z, V, reverse V, and W incisions. In our experience, meticulous inverted V midcolumellar incision closure results in a better scar. There is no convincing reason to reject the transcolumellar incision for external rhinoplasty. PMID- 19553832 TI - Reconstruction of growing skull fracture with in situ galeal graft duraplasty and porous polyethylene sheet. AB - OBJECTIVE: In growing skull fractures with large calvarial defects, it is difficult to use autografts for reconstruction and it requires alternative materials for cranioplasty. In this report, the authors describe their experience and introduce reconstruction of the growing skull fractures' defects with a porous polyethylene sheet (Medpor) and with a novel technique of duraplasty with in situ galeal graft, which avoid the potentially risky dissection and exposure of brain tissue. The goal of this study was to clarify effective surgical methods and to provide the rationale for these techniques. METHODS: We performed this technique on 8 patients with large calvarial defects resulting from growing skull fractures. The skin flap was retracted, leaving the galeal plane adherent to the underlying defect. After removing the bony edges and exposing the underlying retracted dural margins, duraplasty was performed by suturing the galeal tissue left in situ on the defect of the dural margins. Bone reconstruction was performed by placing porous polyethylene sheet (Medpor). CONCLUSIONS: Duraplasty with in situ galeal tissue is a simple, safe, and effective technique to reconstruct dural defects in growing skull fracture, which avoids the risky dissection of the brain tissue. Also, by using Medpor, growing skull fractures can be effectively reconstructed with good cosmetic results. PMID- 19553833 TI - Is transfacial Kirschner wire fixation still indicated in isolated zygomaticomaxillary complex fractures? Retrospective study of 216 cases in CHRU of Lille: epidemiology, therapeutic management, and results. AB - INTRODUCTION: No consensus on both contention and reduction a type in zygomaticomaxillary complex (ZMC) fractures' treatment exists. We tried to evaluate the percutaneous hook reduction method and the transfacial Kirschner wire (K-wire) fixation method in these fractures' treatment. This study also analyses epidemiological data of 4 million inhabitants (Nord-Pas-de-Calais region) in this trauma type. MATERIALS AND METHODS: All the ZMC fractures treated in our department from September 2000 to November 2006 were reviewed retrospectively. The patients were evaluated by clinical and radiologic assessment. Therapeutic data and results were analyzed. RESULTS: We reviewed 216 consecutively isolated ZMC fractures managed in our unit: 39 women and 177 men were included (sex ratio, 1:4.5). Mean age is 33.1 years. Assault is the main etiology before motor vehicle crashes. The average treatment delay is 3.5 days, and the mean delay before surgery is 10.3 days. All of the patients who had surgery underwent percutaneous hook reduction, and 77.3% of reduced fractures needed a contention realized at least by interosseous K-wire fixation in 94.6% of the cases. Reduction results were not satisfying in 5.6% of the patients who had surgery. DISCUSSION: This study aimed to evaluate a method of ZMC fractures treatment and to eventually modify some aspects. We proposed a guideline allowing a main importance to clinical examination and to interosseous transfacial K-wire fixation associated with an open reduction with frontozygomatic and/or infraorbital rim osteosynthesis if mandatory. PMID- 19553834 TI - Do we have to dissect infraorbital nerve from periorbita in orbital floor fracture? AB - The aim if this study was to see whether we have to dissect the infraorbital nerve from periorbita in orbital floor fractures.Orbital floors were reconstructed in 174 patients. Among them, 31 patients were treated with horseshoe-shaped synthetic sheets. Preoperative and postoperative findings including hypesthesia, diplopia, Hertel exophthalmometry, and occurrence of complications were checked.After the subciliary incision through skin and orbicularis oculi muscle, the skin-muscle flap was elevated just superficial to the orbital septum to the arcus marginalis. The periosteum was incised and elevated to expose the fracture site. The orbital branch of the infraorbital artery was identified. The periorbita was not separated from the infraorbital nerve, and the orbital branch of the infraorbital artery was preserved. The sheet was trimmed in horseshoe shape as the cleft fits to the infraorbital groove.On physical examination before surgery, diplopia was the most common (58.1%) associated complication, followed by hypesthesia (35.5%), limited ocular movement (9.7%), enophthalmos (3.2%), and hematoma (3.2%).Even if the fracture site is the posterior half, the periorbita does not have to be separated from the infraorbital nerve to avoid injury of the orbital branch of the infraorbital artery. Instead, the fracture site might be covered by a horseshoe-shaped sheet. PMID- 19553835 TI - Analysis of orbital bone fractures: a 12-year study of 391 patients. AB - This retrospective study evaluates 391 patients with orbital bone fractures from a variety of accidents that were treated at the department of Plastic and Reconstructive Surgery, Inha University Hospital, Incheon, South Korea, between February 1996 and April 2008. The medical records of these patients were reviewed and analyzed to determine the clinical characteristics and treatment of the orbital bone fractures.The following results were obtained. The mean age of the patients was 31.1 years, and the age range was 4 to 78 years. The most common age group was the third decade of life (32.5%). There was a significant male predominance in all age groups, with a ratio of 4.43:1. The most common etiology was violent (assault) or nonviolent traumatic injury (57.5%) followed by traffic accidents (15.6%) and sports injuries (10.7%).The most common isolated orbital bone fracture site was the orbital floor (26.9%). The largest group of complex fractures included the inferior region of the orbital floor and zygomaticomaxilla (18.9%). Open reduction was performed in 63.2% of the cases, and the most common fracture reconstruction material was MEDPOR (56.4%) followed by a resorbable sheet (41.1%). The postoperative complication rate was 17.9%, and there were no statistically significant differences among the reconstruction materials with regard to complications. During follow-up, diplopia, hypoesthesia, and enophthalmos occurred as complications; however, there was no significant difference between porous polyethylene sheet (MEDPOR) and resorbable sheet groups.Long-term epidemiological data regarding the natural history of orbital bone fractures are important for the evaluation of existing preventative measures and for the development of new methods of injury prevention and treatment. PMID- 19553836 TI - Long-pulsed Nd: YAG laser treatment in vascular lesions of the oral cavity. AB - Yet no universally accepted treatment protocol for oral hemangiomas and vascular malformations exists. This study determines the long-term clinical outcome after long-pulsed Nd:YAG laser treatment in 121 previously untreated patients (78 females, 43 males; mean age, 19 years) with oral hemangioma or vascular malformations. The end point was 100% vessel clearing after 1 to 3 sessions.The power of the laser was set at 6.5 W per pulse, the pulse widths ranged between 30 and 60 milliseconds. With an optical fiber diameter of 600 microm, the wavelength was constantly 1064 nm. Dynamic cooling device was set at 10 to 20 and 10 to 15 times before and after pulse, respectively. Whereas 77% of lesions were cleared totally after a single session only, 23% required an overall of 2 to 3 sessions. Tissue sloughing occurred in all patients. The mean follow-up period of 13 months (minimum, 6 months; maximum, 24 months) showed neither functional nor cosmetic shortcomings. Long-pulsed Nd:YAG laser treatment proved to be an effective and valuable method for the treatment of oral hemangiomas and vascular malformations. PMID- 19553837 TI - Incidents malignant neoplasias maxillofacial area. AB - Cancer is regarded as abnormal cellular multiplication; it is not controlled by the organism, and its cells present a differentiated DNA. Initially, the disease does not show clinical signs, but it can be diagnosed by laboratory examinations. When tumors are present in the maxillofacial area, the carrier can lose structures in this area, resulting to the carrier's social environment exclusion. This article aims to show incidences and causes of malignant neoplasias in the maxillofacial area. PMID- 19553838 TI - Tongue reduction in Beckwith-Wiedemann syndrome with CO(2) laser. AB - Beckwith-Wiedemann syndrome is a congenital syndrome with some anomaly in overgrowth. Most common manifestations are exomphalos, macroglossia, gigantism, and visceromegaly. Overgrowth in tongue's size caused clinical symptoms such as dysphagia, speech disorder, strong in chewing, upper-airway obstruction, and psychological problems with appearance. Cold surgical techniques are commonly used in treating macroglossia. We presented tongue reduction with laser and its early result on a child with Beckwith-Wiedemann syndrome for macroglossia. PMID- 19553839 TI - Primary hydatid cyst of the infratemporal fossa. AB - Hydatid cyst disease is a rare parasitic disease caused by the larval stage of Echinococcus. This parasite in the larval stage can thrive in many parts of the body, most frequently in the liver. Head and neck involvement of the disease is rare. Herein, we present a case of primary hydatid cyst occurring in the infratemporal fossa, which is an extremely rare localization. PMID- 19553840 TI - The nasal septum: an unusual presentation of metastatic renal cell carcinoma. AB - A 45-year-old woman with metastatic renal cell carcinoma to the nasal septum was presented in this study. Metastatic neoplasms of the paranasal region are so rare that they have been usually reported as case reports. Although there are reported cases of renal cell carcinoma to the paranasal sinuses, this case was unique in that it is an isolated metastasis to the nasal septum. The clinical presentation, radiologic and pathologic investigations, and the treatment of patient with surgery and chemotherapy with sunitinib malate (Sutent) were discussed. PMID- 19553841 TI - Psammomatoid juvenile cemento-ossifying fibroma of the maxilla. AB - Both the trabecular and the psammomatoid juvenile cemento-ossifying fibromas (COFs) are specific entities among the COFs being distinguished because of their age predilection, site involvement, and clinical behavior. A case of a large-size juvenile COF of histologic psammomatoid pattern is shown. The clinicoradiographic and histologic findings as well as the treatment strategy are presented and discussed. PMID- 19553842 TI - Cranial fasciitis presenting as a frontonasal mass. AB - We report the unusual case of a newborn baby with cranial fasciitis of the frontonasal area. Herein, we present the clinical, pathologic, and radiologic findings of a male neonate with cranial fasciitis of the skull. Cranial fasciitis is a benign fibroblastic tumor related to nodular fasciitis, but it has a predilection for the scalp of children. One case has been described in neonates (Neurosurgery. 2001;48:430-435). PMID- 19553843 TI - Reconstruction of the zygomatic arch using a mandibular adaption plate. AB - Zygomatic complex fractures are one of the most common facial fractures treated by maxillofacial surgeons. Because of the importance of the zygomatic complex in midfacial skeletal orientation, comminuted zygomatic arch fractures can lead to significant morbidity and deformity. Common repair modalities include closed reduction via a Gilles approach, open reduction and internal fixation with screws and/or plating systems, primary bone grafting, and/or biosynthetics. In patients with significant comminution and arch defects, who are poor candidates for bone grafting, reconstruction can be difficult. In this clinical report, we present 2 patients with comminuted zygomatic arch fractures and significant bony defects, who were reconstructed with mandibular adaption plates alone without primary bone grafting. Over a mean follow-up period of 12.5 months, both patients healed well without any complications and had acceptable cosmetic outcomes. PMID- 19553844 TI - Conservative treatment of retropharyngeal and parapharyngeal abscess in children. AB - Parapharyngeal and retropharyngeal infections, which can potentially cause life threatening complications, could be treated conservatively with no need for surgical drainage. A retrospective analysis of all patients diagnosed with retropharyngeal and parapharyngeal infections was performed. Information regarding age, sex, presenting symptoms, physical examination, laboratory and imaging evaluations, management, duration of hospital stay, and complications were reviewed. Seven children were identified, 4 with retropharyngeal abscess and 3 with parapharyngeal abscess. All but 1 patient were under 7 years old, and all were treated with intravenous amoxicilin/clavulanic acid and corticosteroids. Torticollis and fever were present in all the patients. The mean length of hospital stay was 7 days. There were no complications associated. We demonstrate that retropharyngeal and parapharyngeal abscesses can be treated medically, reserving the surgical drainage for complicated cases. Treatment with intravenous antibiotics and corticosteroids is a safe option, reducing the durations of symptoms and the length of hospital stay. PMID- 19553845 TI - Concha bullosa: endoscopic treatment. AB - AIM: To assess the frequency and the endoscopic treatment of the middle turbinate pneumatization or concha bullosa. MATERIALS AND METHODS: Forty-nine patients (26 males and 23 females) with sinusitis and headache symptoms and axial and coronal computed tomographic scans of the paranasal sinuses and who had an endoscopic examination visit in the outpatient department between January 2005 and July 2007 were included in this study. Functional endoscopic sinus surgery was performed. Nasal tampons were removed 3 days after surgery, and endoscopic examination visits were performed 7, 15, 30, and 60 days after surgical treatment. RESULTS: All the patients presented complaints of chronic nasal obstruction. Eleven patients (5 males and 6 females; 22.4%) presented either a unilateral or a bilateral middle turbinate pneumatization. The surgeons recorded the surgery and each examination visit. In remote controls (12 mo at least), the patients showed a total remission of symptoms. There were no important complications. PMID- 19553846 TI - Fabrication of an auricular prosthesis using computed tomography and rapid prototyping technique. AB - This article describes the use of computed tomography and rapid prototyping method to produce a wax pattern of a missing ear. Computed tomographic scans were obtained from the patient, and a three-dimensional digital image was produced using a computer-aided design software. Then, the image of the present ear was extracted, and its mirror image was obtained with the software. The pattern of the inverted three-dimensional cast was produced using a rapid prototyping machine. The cast was duplicated in wax of identical dimensions and shape of the opposing ear, and the auricular prosthesis was fabricated with traditional methods. PMID- 19553847 TI - Intraglandular toxoplasmic lymphadenitis of the parotid gland. AB - Most preauricular masses are parotid neoplasms; however, some infectious and inflammatory causes may exhibit similar presentation. Toxoplasmosis is a worldwide parasitary disease. The clinical presentation of toxoplasmosis is mostly asymptomatic and may include subfebrile fever, fatigue, and lymphadenopathy. Only 13 clinical reports of intraglandular toxoplasmic lymphadenitis have been previously reported in the English literature. A careful history and examination with appropriate investigations, including immunoglobulin G avidity assay, will frequently provide the diagnosis of intraparotid toxoplasmosis. Surgery might therefore have been delayed or avoided depending on a satisfactory clinical improvement in consequence of proper medical treatment. PMID- 19553848 TI - Ameloblastoma relapse after 50 years from resection treatment. AB - Ameloblastoma is a slow-growing, locally invasive, epithelial odontogenic tumor of the jaws with a high rate of recurrence if not removed adequately but with virtually no tendency to metastasize (World Health Organization Classification of Tumors: Pathology and Genetics of Head and Neck Tumours, 2005). This paper presents a case of a woman who was treated in 1961, when she was 25 years old, for an ameloblastoma in the right posterior region of the mandible. After 50 years, the ameloblastoma relapsed, and another surgical treatment was necessary. PMID- 19553849 TI - Fibrovascularization and osteogenesis in high-density porous polyethylene implants. AB - High-density porous polyethylene (HDPP) has been extensively used in craniofacial reconstructions with high-level success and minimal complications. It is known for its biocompatibility and satisfactory stability in the receptor bone area, presenting only a few reports of mobility and infection. In the current study, attention was given to the interface area between HDPP and bone surface to analyze fibrous and bone tissue formation and ingrowth into the pores of the material placed in the mandible of rabbits. Twelve male New Zealand rabbits underwent surgical procedure to receive bilateral HDPP implants in buccal face of dentate mandibular alveolar process, fixed with titanium screws. After 7, 14, 45, and 90 days, the animals were killed, and the specimens were retrieved for histologic and immunohistochemical analyses. No implant loss or infection was detected at the retrieval of the specimens. The microscopic analysis presented satisfactory integration of the material to the bone surface, with new bone formation from the receptor bed and inside the pores of the material, observed from the 15th day. After 90 days, remodeling bone and fibrous tissue was seen in the interface region. Among some of the pores, mature lamellar bone was present. Immunohistochemistry pointed out a moderate expression either to Core binding factor protein 1/RUNX2 or to vascular endothelial growth factor for early periods evaluated, that is, 7 and 15 days after surgery. These results confirm the osteoconductive behavior and high biocompatibility of the material, associated to its adequate immobilization, leading to its lifelong presence in human biologic system. PMID- 19553850 TI - Carcinoma of the buccal mucosa metastasizing to the talus. AB - Head and neck squamous cell carcinomas (HNSCCs) are tumors with propensity mostly for locoregional spread. Most frequent sites of metastasis from cancer of the oral cavity include lung, bone, liver, adrenal, heart, and kidney. The advent of newer surgical techniques and an improved understanding of head and neck cancer have further improved control of cancer above the clavicles. Furthermore, the overall survival rate in patients with advanced head and neck cancer has not improved significantly. This has been partly because of the emergence of second primary cancers and the development of distant metastasis defined as cancer deposits below the clavicles. Authors reported a case of a 60 years-old man affected by squamous cell carcinoma of the oral cavity treated with surgery and adjuvant radiochemotherapy that developed a metastatic lesion to the left talus 10 months after primary treatment. In conclusion, bone is a rare metastatic site for HNSCC, much rarer in case of sites distal to the knee. Owing to paucity of the literature on this, no definite predisposing factor is identifiable. Because of the rarity of these lesions, bone scintigraphy remains not indicated in case of HNSCC, but attention has to be paid to eventual symptoms, also if frequently sensitive for advanced lesions. PMID- 19553851 TI - Accuracy and predictability in use of AO three-dimensionally preformed titanium mesh plates for posttraumatic orbital reconstruction: a pilot study. AB - The aim of this study was to prospectively evaluate the accuracy and predictability of new three-dimensionally preformed AO titanium mesh plates for posttraumatic orbital wall reconstruction.We analyzed the preoperative and postoperative clinical and radiologic data of 10 patients with isolated blow-out orbital fractures. Fracture locations were as follows: floor (N = 7; 70%), medial wall (N = 1; 1%), and floor/medial wall (N = 2; 2%). The floor fractures were exposed by a standard transconjunctival approach, whereas a combined transcaruncular transconjunctival approach was used in patients with medial wall fractures. A three-dimensional preformed AO titanium mesh plate (0.4 mm in thickness) was selected according to the size of the defect previously measured on the preoperative computed tomographic (CT) scan examination and fixed at the inferior orbital rim with 1 or 2 screws. The accuracy of plate positioning of the reconstructed orbit was assessed on the postoperative CT scan. Coronal CT scan slices were used to measure bony orbital volume using OsiriX Medical Image software. Reconstructed versus uninjured orbital volume were statistically correlated.Nine patients (90%) had a successful treatment outcome without complications. One patient (10%) developed a mechanical limitation of upward gaze with a resulting handicapping diplopia requiring hardware removal. Postoperative orbital CT scan showed an anatomic three-dimensional placement of the orbital mesh plates in all of the patients. Volume data of the reconstructed orbit fitted that of the contralateral uninjured orbit with accuracy to within 2.5 cm(3). There was no significant difference in volume between the reconstructed and uninjured orbits.This preliminary study has demonstrated that three-dimensionally preformed AO titanium mesh plates for posttraumatic orbital wall reconstruction results in (1) a high rate of success with an acceptable rate of major clinical complications (10%) and (2) an anatomic restoration of the bony orbital contour and volume that closely approximates that of the contralateral uninjured orbit. PMID- 19553852 TI - A three-dimensional analysis of the relationship among lower facial width, bony width, and masseter muscle volume in subjects with prominent mandible angles. AB - Despite the fact that a face with a square appearance is determined by surface anatomic features, skin surface parameters and their relationship with inner anatomic features have not been evaluated owing to the challenge of accurately and consistently measuring skin landmarks.We introduce 2 surface distances of the lower face obtained from images of skin and bone thresholds from three dimensional computed tomographic scans; these were realigned in identical positions using the Frankfurt horizontal plane. The selected parameters were skin surface (LFOP) and bony width (MOP) of the occlusal plane level, skin surface (LFBP) and bony width (MBP) of the bigonial plane level, masseter volume (MV), and soft tissue thickness. Ten subjects with mandible angle flaring and 10 control subjects without flaring were evaluated.The parameters LFOP, LFBP, MBP, and MV showed differences between the study and control groups (P < 0.05). Lower facial width of the occlusal plane was longer than LFBP in both groups (P < 0.005), and MOP was shorter than MBP in the study group (P < 0.005), whereas MOP was greater than MBP in the control group (P < 0.001). Correlation analysis revealed that skin surface width was significantly related to bony width only in the control group (r > 0.6). Masseter volume showed no significant relationship with any skin surface or bony parameter but with soft tissue thickness in the control group (r > 0.6).In conclusion, skin surface widths (LFOP and LFBP) along with MV and bony width differ between patients with flared and nonflared mandibles. Our findings suggest that the skin surface width of the lower face can be used as a valuable landmark. PMID- 19553854 TI - Stereolithographic volume evaluation of healing and shaping after rhinoplasty operations. AB - Nasal edema and volume changes are unavoidable processes during the healing period after rhinoplasty. Various applications were reported regarding the prevention of early edema; however, the literature shows no study focused on the course of the nasal edema and volume changes up-to-date. We aimed to study the nasal volume changes during the first year of postoperative healing period and to form a recovery and volume change diagram with the obtained data. We prepared standard frames and nasal molds of 7 rhinoplasty patients at regular time intervals (preoperative period and at the postoperative 1st, 2nd, 4th, 8th, 12th, 24th, and 52nd weeks). Plaster nasal models were created by using these molds. Volumes of models were measured by computed tomographic scanning and three dimensional image processing programs. According to our results, the nasal edema reaches its maximum level at the postoperative fourth week and then rapidly decreases until its minimum level at the eighth week. In contrast with the general opinion, the nasal volume begins to increase smoothly reaching to a level minimally below the preoperative value by the end of the first year. PMID- 19553853 TI - Finite element analysis to compare complete denture and implant-retained overdentures with different attachment systems. AB - This finite element analysis compared stress distribution on complete dentures and implant-retained overdentures with different attachment systems. Four models of edentulous mandible were constructed: group A (control), complete denture; group B, overdenture retained by 2 splinted implants with bar-clip system; group C, overdenture retained by 2 unsplinted implants with o'ring system; and group D, overdenture retained by 2 splinted implants with bar-clip and 2 distally placed o'ring system. Evaluation was performed on Ansys software, with 100-N vertical load applied on central incisive teeth. The lowest maximum general stress value (in megapascal) was observed in group A (64.305) followed by groups C (119.006), D (258.650), and B (349.873). The same trend occurred in supporting tissues with the highest stress value for cortical bone. Unsplinted implants associated with the o'ring attachment system showed the lowest maximum stress values among all overdenture groups. Furthermore, o'ring system also improved stress distribution when associated with bar-clip system. PMID- 19553855 TI - Intraoperative repositioning assessment using navigation system in zygomatic fracture. AB - BACKGROUND: Intraoperative repositioning assessment in the zygomatic fracture operation is very important in achieving adequate facial symmetry. A navigation system that has been developed in neurosurgery has recently been applied in the otolaryngol, orthopedic, and maxillofacial surgeries. We used a navigation system (Stealth Station TREON; Medtronic) in zygomatic fracture operation, and found it useful in performing accurate reduction and confirming the symmetry. METHODS: We applied a navigation system in 6 patients with zygomatic bone fracture. Navigation aided reduction of the bone fracture was performed by evaluating intraoperative procedures: (1) an evaluation by measuring the distances from the midline of the face and (2) an evaluation by comparing the affected side to the mirror image of the nonaffected side. A method using the distance from the midline of the face was performed in 3 cases, and a method using the mirror image of the nonaffected side was used in 6 cases. RESULTS: Accurate reduction and symmetry were confirmed during the operation in all patients. Good symmetry was confirmed in postoperative computed tomographic scan. It was useful in performing accurate reduction of the fractures and confirming the symmetry of the face. CONCLUSIONS: We suggest that the system can also be applied for the treatment of untreated facial fractures requiring osteotomy and of comminuted facial fractures with third-bone fragments and for the evaluation of bone graft. PMID- 19553856 TI - Safety profile of wire osteosynthesis in craniosynostosis surgery. AB - With the advent of resorbable systems, most surgeons have stopped using wires for craniofacial fixation. Although numerous large retrospective reports regarding craniofacial surgery have been published, no documentation exists regarding the disadvantages or complications associated with wires. We review our experience with 47 consecutive patients with bicoronal and unicoronal craniosynostosis where wire osteosynthesis alone was used. Nine patients (19.1%) developed wire-related complications, but only 5 patients (10.6%) required reoperations. No other complications were observed including growth restrictions, implant migration, or interference with radiographic imaging. These results are comparable to those reported in the literature for other fixation systems and demonstrate that wires are a safe means of fixation of the cranial vault in infancy. PMID- 19553857 TI - Treatment of lymphatic malformations with OK-432 (Picibanil): review of the literature. AB - INTRODUCTION: Lymphatic malformations (LM) are benign structural defects that can cause serious complications because of their size and location. Traditionally, surgical removal was the first treatment modality, but this could be associated with many complications and risks. Since Ogita introduced OK-432 (picibanil) in 1987 as a treatment method, this sclerosant has become popular. This paper is a review of the trials published so far on this topic. PATIENTS AND METHODS: A literature search of English trials with 5 or more patients in it with LM who had never been treated before was done. The paper had to use the microcystic macrocystic classification and have a mean follow-up of more than a year to be included in this review. Results were classified as "excellent" when the lesions show a regression of more than 90%, "good" when regression is more than 50%, and "poor" when shrinkage is less than 50% (this also includes no response at all). RESULTS: Twenty-seven percent of microcystic LMs show an excellent result; 33%, a good result; and 40%, a poor result. Of the macrocystic LMs, 88% have excellent results. Recurrence rates vary from 5% to 8%. The adverse effects are mostly mild. DISCUSSION: Most trials have a short follow-up; therefore, there are uncertainties when it comes to cure and regression. Mostly, the adverse effects of OK-432 are trivial and disappear after a week, but the need for a temporary tracheostomy has been described. Screening for allergic reactions to penicilline is needed, with the risk of anaphylactic shock in mind. It is difficult to compare the different techniques used by the authors, and none of the trials included in this study are randomized controlled trials; most are retrospective and were so-called level 4 studies. CONCLUSIONS: This review demonstrates that OK 432 is an effective way to treat LM. Because of a possible risk of airway obstruction, treatment should always take place in specialized treatment facilities. Macrocystic lesions show a better response to OK-432 treatment than microcystic lesions. Serious complications with OK-432 are infrequent, and this type of sclerotherapy seems to have no influence on future surgery. We therefore suggest the use of OK-432 as an effective first-line treatment of LMs. PMID- 19553858 TI - Evaluation and management of pediatric hand injuries resulting from exercise machines. AB - Traumatic injury to the hand in the pediatric population unfortunately remains common in the spectrum of accidental injuries sustained in this population. This is particularly true regarding stationary exercise equipment. The popularity of home exercise equipment continues to rise in the United States, particularly stationary bicycles and treadmills. Additional safety mechanisms are required to prevent the anticipated increase in the number of hand injuries to children as a result of these machines. The aim of this paper was to review the incidence and management of pediatric hand injuries sustained as a result of stationary exercise equipment in the home, in addition to increasing awareness of these types of injuries. PMID- 19553859 TI - Fingertip-nail bed injuries in children: current concepts and controversies of treatment. AB - Crush injuries of the fingertip are the most common hand injuries seen in children. Many involve fracture of the distal phalanx, whereas others result in either crush alone or complete or partial fingertip amputation. The need for nail removal and nail bed repair after crush injury to the fingertip has long been a matter of debate. In our study comparing the outcome of nail removal and formal nail bed reconstruction versus simple evacuation of the subungual hematoma via trephination after fingernail crush injuries, simple nail trephination was equal to, or superior to, removal of the nail and nail bed repair with significantly lower cost (Roser SE, Gellman H. Comparison of nail bed repair versus nail trephination for subungual hematomas in children. J Hand Surg [Am] 1999;24:1166 1170). PMID- 19553860 TI - Embryology of the hand and upper extremity. AB - As a universal means of communication and a critical tool for survival, the human hand is of extraordinary importance to our evolutionary survival. As the product of countless overlapping chemical signals, the upper extremity is highly dependent on a multifactoral web of genetic and environmental factors. At the molecular level, specialized signaling centers guide limb development along 3 spatial limb axes: (1) proximodistal, (2) anteroposterior, and (3) dorsoventral. Within the growing limb bud, the 3 main signaling centers are (1) the apical ectodermal ridge, (2) the zone of polarizing activity, and (3) the nonridge ectoderm. Cells within these signaling centers govern the process of limb differentiation via secretion of various chemical messengers. Although each aspect of extremity growth seems directly dependent on a process-specific mechanism, overall limb development relies on the proper interaction of these countless protein factors. Here, we review the macroscopic development of the upper limb and discuss the complex mechanisms underlying differentiation of the human hand. PMID- 19553861 TI - Constriction ring syndrome. AB - Constriction ring syndrome is a congenital anomaly with sometimes devastating consequences. Because of the unknown etiology, treatment is aimed at recovery of function and prevention of the sequelae rather than prevention of the syndrome. This paper aimed to review the theories of the etiology, the features and associated defects, and the operative management of constriction ring syndrome. PMID- 19553862 TI - Peripheral nerve injuries of the pediatric hand: issues in diagnosis and management. AB - Peripheral nerve injuries resulting in significant neural disruption frequently present complex management challenges. Typically the product of fracture, dislocation, or crush injuries, pediatric peripheral nerve injuries may be difficult to accurately characterize. Thorough clinical examination coupled with electromyogram and neurophysiologic studies are extremely useful. When possible, primary repair should be attempted. If, however, defect size precludes primary reanastomosis, use of a nerve graft may be advantageous. Alternatively, nerve conduits, such as veins, pseudosheaths, and bioabsorbable tubes, are also effective facilitators of nerve regeneration. Although nerve injuries of the pediatric hand often present complex challenges, a thorough knowledge of diagnostic methods and advances in surgical interventions offers better outcomes. PMID- 19553863 TI - Understanding the prevalence of inpatient falls associated with toileting in adult acute care settings. AB - This qualitative study determined the prevalence of inpatient falls that were associated with toileting in a Michigan community hospital. Of all falls, 45.2% were related to toileting. The most common theme was falling on the way from the bed or chair to the bathroom. Nurses should focus on safe patient transfers and on using the completed risk assessment and should develop an individualized prevention plan for each patient based on their needs. PMID- 19553865 TI - Has there been progress in schizophrenia research and treatment for all peoples during 2008? Existing disparities in mental healthcare and research? PMID- 19553866 TI - Language pathway abnormalities in schizophrenia: a review of fMRI and other imaging studies. AB - PURPOSE OF REVIEW: Schizophrenia is a severe mental disorder with substantial genetic vulnerability. This review discusses recent neuroimaging studies reporting on impairment in brain functioning relevant to language processing in individuals with schizophrenia and those who are at a genetic risk for its development. RECENT FINDINGS: Studies have shown that schizophrenia is associated with deficits in language function, as well as structural and functional abnormalities in brain regions that are involved with language perception and processing. Individuals who are at genetic high risk for schizophrenia also have structural and functional deficits in brain pathways for language processing. These studies consistently suggest that the normal pattern of left hemisphere dominance of language processing is significantly disturbed. SUMMARY: This review suggests that future studies should examine the underlying mechanism for producing this disturbance in language processing and that prospective studies should be carried out that aim to follow individuals over time to determine whether these anomalies eventually lead to clinical symptoms of schizophrenia. PMID- 19553867 TI - Emotional face processing in schizophrenia. AB - PURPOSE OF REVIEW: Recent studies of face processing deficits in people with schizophrenia have begun to shed light on the answers to four important questions: What is the major component of the deficit? Does it have a neurobiological basis? When does it appear in development? Can it be remediated? RECENT FINDINGS: Eye contact is especially important for recognizing fearful expressions. Patients with schizophrenia avoid gazing at important facial feature regions (especially eyes) and are particularly impaired in recognizing fear. Patients were thought to exhibit decreased amygdala activity when viewing fearful faces; however, more recent studies have revealed limbic hyperactivity in patients when viewing nonfearful and fearful faces compared with baseline. Amygdala hyperactivity can also be detected when people with schizophrenia fail to appropriately recognize fear faces. Studies indicate that there are developmental changes in amygdala activation, including limbic hyperactivity to fear faces during adolescence, a critical time for the onset of schizophrenia. New treatment strategies can increase gaze to the eye region, which could improve emotional recognition in patients. SUMMARY: These findings suggest that there may soon be more options for overcoming specific problems in emotional face evaluation in people who have schizophrenia. PMID- 19553868 TI - Update on key previously proposed candidate genes for schizophrenia. AB - PURPOSE OF REVIEW: We will give an overview of more recent data concerning previously implicated candidate genes for schizophrenia. This includes functional data when available. Furthermore, studies on copy number repeats and their possible implications in schizophrenia will be described. RECENT FINDINGS: Within the past year, schizophrenia genetics has focused on a more detailed investigation of previously implicated candidate genes. In addition, investigation of copy number variations has led to the identification of rare structural DNA variants that might play a major role in some cases of schizophrenia. SUMMARY: There is emerging evidence that some cases of schizophrenia might be due to rare genetic structural variation, though the majority of cases should be due to a cumulative effect of common variations in multiple genes, which in combination with environmental stressors may lead to the development of schizophrenia. PMID- 19553869 TI - Recent advances in postmortem pathology and neurochemistry in schizophrenia. AB - PURPOSE OF REVIEW: This is a review examining recent data from the study of the postmortem central nervous system (CNS) of patients with schizophrenia. RECENT FINDINGS: Studies on the human CNS transcriptome suggest changes in pro inflammatory pathways and myelination in schizophrenia, whereas changes in the proteome suggest that pathways involved in energy and metabolism may be particularly stressed. There appear to be complex changes in the expression of proposed candidate genes for schizophrenia such as NRG1, DISC1, RGS4 and DTNB1, and there are continued reports of alterations in central gamma-aminobutyric acidergic, dopaminergic, glutamatergic and cholinergic pathways in patients with the disorder. Data on epigenetic mechanisms and transcriptome regulation suggest that at least some changes in gene expression may be due to changes in levels of gene promoter methylation or microRNAs in the CNS of patients with schizophrenia. SUMMARY: Postmortem CNS studies have begun to unravel changes in the epigenetic regulation of gene expression that may be central to how gene-environment interactions contribute to the onset of schizophrenia. In addition, a recent study indicates that it is possible to use biomarkers to segregate the syndrome of schizophrenia into more biologically homogeneous populations, which should decrease the biological complexity observed within that group within the schizophrenia syndrome. PMID- 19553870 TI - Cognitive remediation for schizophrenia: it is even more complicated. AB - PURPOSE OF REVIEW: Attempts to remediate the cognitive difficulties of people with a diagnosis of schizophrenia have shown efficacy; however, existing treatment studies display huge variation, frustrating efforts to determine how best to apply this treatment tool. This review summarizes findings of induced cognitive change in schizophrenia, with reference to the remediation method, the presence of accompanying treatment(s), how cognitive change generalizes and its value. RECENT FINDINGS: Although there is strong evidence that cognitive change can be induced in schizophrenia, there is little evidence for the superiority of any treatment approach. Furthermore, remediation is most effective when in combination with other treatments, such as vocational training. Cognitive rehabilitation can be cost-effective and is valued by patients. A number of studies show generalization of positive outcomes beyond cognitive variables, with more focused treatment associated with less generalization. SUMMARY: Induced cognitive change does not necessarily need to be large to facilitate functional outcomes. Instead, opportunities to apply newly acquired cognitive skills and strategies in the real world are vital. The next generation of studies need to compare different treatments using specific and more general measures of cognitive outcome. These may then throw light on their mechanisms of action. The results of these more sophisticated studies will allow therapists to tailor treatments to individuals to maximize gain for patients. PMID- 19553871 TI - Recent diffusion tensor imaging findings in early stages of schizophrenia. AB - PURPOSE OF REVIEW: Several lines of evidence suggest that the normal integration of cerebral function may be compromised in schizophrenia. Abnormalities in white matter tracts, which connect brain regions into functional networks, may be directly relevant to its pathophysiology. Diffusion tensor imaging (DTI) has increasingly been used to study white matter abnormalities in schizophrenia; in this review, we will discuss recent DTI findings focusing on the early stages of the disorder. RECENT FINDINGS: Deficits in white matter integrity as inferred by DTI appear to be present in the early stages of schizophrenia, even in neuroleptic-naive patients, and may be the result of interaction between illness related processes and normal development. The pattern of identified abnormalities is not totally consistent across all studies, with frontotemporal, frontoparietal and temporooccipital connections as well as projection fibers and cerebellar white matter being among the affected tracts. SUMMARY: Recent DTI findings further support the hypothesis of structural dysconnectivity in schizophrenia. The presence of white matter abnormalities early in the course of the illness is suggestive of these being related to the emergence of the disorder. PMID- 19553872 TI - Intervention in the at-risk state to prevent transition to psychosis. AB - PURPOSE OF REVIEW: The number of intervention studies aiming to prevent psychosis is still small. Follow-up data of the first studies were published during the last year and neuroprotection has become an important issue. RECENT FINDINGS: Initially superior effects of pharmacological or cognitive intervention reported by the first studies in the field became less clear about 3 years after cessation of intervention; however, a common problem of these first trials is a small sample size resulting in a lack of sufficient statistical power. The first studies of interventions thought to act as primarily neuroprotective yielded promising findings; however, further studies are needed to evaluate the preventive as well as the neuroprotective efficacy of these approaches. SUMMARY: Besides methodologically sound studies, improved enrichment strategies are required as well as risk-adapted intervention strategies, guided by evidence based clinical staging algorithms. Furthermore, the current concept of psychosis prevention, requiring an intervention to show long-lasting effects even after cessation, needs reconsideration. Approaches as used for relapse prevention in psychosis or for chronic at-risk states in internal medicine may help to maintain the initial superior prophylactic effects. PMID- 19553873 TI - Role of behavioural and social sciences in medical education. AB - PURPOSE OF REVIEW: It is widely accepted that behavioural and social sciences (BSS) should become an integral component of medical training. This review looks at the extent to which BSS are included in medical curricula and how well they are taught in medical schools across the world. RECENT FINDINGS: Published literature on the role of BSS in medical training is scanty and largely from developed countries. Although the need for including BSS in medical education was recognized more than 30 years ago, only little progress has been made in improving the quality and quantity of behavioural sciences' teaching in medical schools, even in developed countries. Numerous barriers that impede better integration have been identified. There are no uniform guidelines, well established curricula, training modules or materials. There is also a severe shortage of adequately qualified and experienced teachers. SUMMARY: Although the role and significance of BSS in medical education and practice is no longer disputed, the teaching of BSS is still highly problematic all over the world. Concerted efforts by medical educators at various levels are needed to improve the quality and quantity of BSS in medical training. PMID- 19553874 TI - Recent research in stress, coping and women's health. AB - PURPOSE OF REVIEW: To highlight recent publications in the area of stress and coping, with specific reference to women's physical health status. RECENT FINDINGS: The transactional model of stress and coping continues to be the mainstay of research in this area. Several longitudinal studies have demonstrated that stress appraisal and resultant coping responses affect health outcome and health-related quality of life in women. In addition to problem-focused coping, women often use distraction methods, seeking social support and faith or religious coping. Psychological interventions in chronic medical conditions need to move beyond education and incorporate more cognitive behavioral components, at the same time addressing women's specific needs. SUMMARY: Coping behaviors in response to the negative threat appraisal of a chronic or severe medical illness serve to reduce psychological distress. However, it is still not clear how they impact at the physiological level. In addition, coping responses, which enhance positive effects and promote health-related quality of life, merit greater attention from researchers. There is a need for more gender comparative research to improve health outcomes in men and women. PMID- 19553875 TI - Behavioural interventions to reduce the risk of physical illness in persons living with mental illness. AB - PURPOSE OF REVIEW: It is beyond dispute that people living with mental illness suffer a disproportionate disease burden when compared with people in the general population. This review considers the efficacy and effectiveness of lifestyle behavioural interventions at service delivery level as a strategy to reduce the risk factors that contribute to somatic disease comorbidity. As many factors contribute to the very poor physical health of people living with mental illness and as there are no 'quick fix' remedies, strategies to improve physical health need to be sustainable on a system-wide basis. RECENT FINDINGS: Most studies of behavioural interventions at best report modest success during the period of the intervention. However, even limited success can significantly reduce the likelihood of physical comorbidities developing. Unfortunately, the evidence suggests that any gains during the intervention are mostly lost over time. The implication is that interventions need to be sustainable over the long-term. SUMMARY: When planning behavioural interventions, consideration ought to be given to extending them over a period of years not weeks or months. Approaches that include additional on-going support beyond the intervention period itself promote a greater likelihood of maintaining the improved physical health of the target population. PMID- 19553876 TI - Chronic pain syndromes and their treatment by psychological interventions. AB - PURPOSE OF REVIEW: Treatment of chronic pain has become a multidisciplinary endeavour including psychological interventions. Databases for life science journals were searched for citations from 2007 and 2008 to determine the current focus of research and the state of evidence. RECENT FINDINGS: Several reviews on systematic research studies confirm that psychological interventions are efficacious in the treatment of chronic musculoskeletal pain, especially back pain, though effect sizes are small and, in some cases, moderate. Findings from clinical practices and treatment centres corroborate these conclusions. The integration of psychological treatment into primary care has not yet proven its utility. Cost-effective interventions to reduce relapse are currently being examined. Psychological headache treatment has again become a topic of research. Evidence is inconsistent, with improvement ranging from an extraordinary size to none at all. Hypnotherapy in children and adolescents with recurrent gastrointestinal pain, examined in a study of high methodological quality, achieved an exceptional level of symptom relief. The aim of two studies on therapy for fibromyalgia and temporomandibular disorder was the identification of mediators and moderators of treatment outcome. SUMMARY: Regarding different pain syndromes such as chronic back pain, headache, fibromyalgia, and temporomandibular disorder, as well as gastrointestinal pain in children, psychological interventions proved their significance for the achievement of favourable treatment outcome. PMID- 19553877 TI - Recent evidence supports emotion-regulation interventions for improving health in at-risk and clinical populations. AB - PURPOSE OF REVIEW: The regulation of strong emotions has important implications for health, particularly among individuals with chronic illness. We focus this brief review on effective psychosocial interventions that emphasize and teach skills to improve emotion regulation in the context of health-related outcomes. RECENT FINDINGS: Recent work in the area of emotion-regulation interventions has tested the effects of emotion-regulation family therapy, group-based emotion regulation psychotherapy, expressive writing, and school-based prevention programs. Emotion-regulation psychotherapy for families shows some benefits for both patients and their family members. Group emotion-regulation interventions and expressive writing result in physical and psychosocial improvement for patients with medical or psychiatric illness. School-based programs show improved emotion knowledge, emotion regulation, and emotional competence, relative to standard academic curricula and existing prevention programs. SUMMARY: Evidence generally supports the use of a variety of emotion-regulation interventions to improve health and well being in at-risk and clinical populations, although factors related to treatment response warrant additional research. PMID- 19553878 TI - Treatment of depression in diabetes: an update. AB - PURPOSE OF REVIEW: The combination of depression and diabetes is common and especially harmful because depression has a strong impact on psychosocial as well as medical outcomes in patients with diabetes. Consequently, treatment for depression in diabetes is also aimed at improvement in glycemic control and risk reduction for diabetes complications and mortality. This review provides an overview of all published, randomized controlled trials on the treatment of depression in patients with diabetes and summarizes current, ongoing research. RECENT FINDINGS: The best results for medical and psychological outcomes were observed for psychological treatments; however, the generalizability of these results is restricted by methodological limitations. Most antidepressants were effective treatments for depression in diabetes but failed to show benefits regarding diabetes-related medical variables. Algorithm-based care, including psychological and psychopharmacological approaches, provides the best scientific evidence for successful depression treatment but not for glycemic control. SUMMARY: Depression can be treated with antidepressants, psychotherapy or a flexible combination of both with relatively good results that are comparable to those for patients who have depression but not diabetes. Up to now, no single treatment that consistently leads to better medical outcomes in patients with both depression and diabetes has been clearly identified. PMID- 19553879 TI - Psychosocial aspects of genetic testing. AB - PURPOSE OF REVIEW: With rapid advances in genetic testing for disease susceptibility, behavioral medicine faces significant challenges in identifying likely patterns of use, how individuals interpret test results, and psychosocial and health impacts of testing. We review recent research on these psychosocial aspects of genetic testing for disease risk. RECENT FINDINGS: Individuals exhibit limited sensitivity in their perceptions of genetic risk information, and mental representations of disease risk appear to guide testing perceptions and behavioral responses. Motivations to undergo testing are complex, and efforts to develop decision aids are underway. Findings on psychological and behavioral impacts of genetic testing vary markedly, with some evidence of minimal or positive effects and other evidence indicating negative consequences that may be undetectable using common measures of general well being. Recent evidence suggests that genetic risk information can motivate health behavior change. Research demonstrates wide-ranging influences of testing on family dynamics, and use of genetic testing with children is of increasing concern. SUMMARY: More research is needed to determine how to structure health communications and counseling to motivate informed use, promote positive responses, and optimize behavior change. Given the ramifications of genetic information for families, personalized genomics will demand a shift toward a family-based healthcare model. PMID- 19553881 TI - Current world literature. PMID- 19553880 TI - Brain dysfunction behind functional symptoms: neuroimaging and somatoform, conversive, and dissociative disorders. AB - PURPOSE OF REVIEW: Neuroimaging research in psychiatry has been increasing exponentially in recent years, yet many psychiatrists are relatively unfamiliar with this field. This article summarizes the findings of the most relevant research articles on the neuroimaging of somatoform, conversive, and dissociative disorders published from January 2007 through June 2008. RECENT FINDINGS: Neuroimaging findings summarized here include alterations of stress regulation and coping in somatoform pain disorders, the importance of catastrophizing in somatization disorder, and the relevance of a history of physical/sexual abuse in irritable bowel syndrome. Regarding fibromyalgia, three of the most significant advances have been the impossibility of differentiating primary and concomitant fibromyalgia in the presence of quiescent underlying disease, the role of hippocampal dysfunction, and the possibility that fibromyalgia may be characterized as an aging process. In dissociative disorders, the high levels of elaborative memory encoding and the reduced size of the parietal lobe are highlighted. SUMMARY: The most promising clinical consequence of these studies, in addition to improving knowledge about the etiology of these illnesses, is the possibility of using neuroimaging findings to identify subgroups of patients, which could allow treatments to be tailored. PMID- 19553883 TI - Sonochemical preparation of polymer nanocomposites. AB - This review covers sonochemical fabrication of polymer nanocomposites. In addition to its application to the synthesis of various polymeric systems, due to its powerful efficiency, sonochemistry has been widely used not only as the assistant of dispersion for nanomaterials such as carbon nanotubes (CNT) and organophillic clay, but also as a special initiator to enhance polymerization for fabrication of polymer nanocomposites with CNT and metallic nanoparticles. Recent developments in the preparation of multi-walled carbon nanotube/polymer nanocomposites with polystyrene and PMMA, magnetic particle/CNT composites and polymer/clay nanocomposites along with their physical characteristics and potential engineering applications will be introduced. Physical characterizations include morphological, thermal, and rheological properties under either an applied electric or magnetic field. PMID- 19553884 TI - Synthesis of a diamino substituted terphenyldivinyl chromophore. AB - (E,E)-1,4-bis(4'-aminostyryl)-2,5-bis(octyloxy)-benzene (6) and its derivative (E,E)-1,4-bis(4'-acetamidostyryl)-2,5-bis(octyloxy)-benzene (7) were synthesized and characterized after alkylation, bromomethylation, Horner-Emmons reaction and reduction from hydroquinone. In order to gain more molecular electronic data, HOMO and LUMO of compound 6 have been calculated by Gaussian 03 W. PMID- 19553885 TI - Highly efficient esterification of ferulic acid under microwave irradiation. AB - A highly efficient synthesis of alkyl ferulates under microwave irradiation is described. The time of these reactions ranged from 3 to 5 minutes, which was much shorter than the traditional synthetic methods, and the alkyl ferulates were obtained in higher yields. PMID- 19553886 TI - Effect of ionic liquids as the mobile phase additives on the HPLC resolution of four active compounds from Sophora flavescens Ait. AB - The retention behaviour of four active compounds from Sophora Flavescens Ait using three ionic liquids as mobile phase modifiers was examined. The effect of the pH and the amount of ionic liquid modifier on the retention of these compounds was determined in methanol/water (v/v) as the mobile phase containing different ionic liquids ranging in concentration from 0.1 mmol/L to 3.0 mmol/L. The ionic liquids showed promise as additives in high-performance liquid chromatography. PMID- 19553887 TI - 2beta-(Isobutyryloxy)florilenalin, a sesquiterpene lactone isolated from the medicinal plant Centipeda minima, induces apoptosis in human nasopharyngeal carcinoma CNE cells. AB - Centipeda minima is a medicinal plant reputed in China as a remedy for nasopharyngeal carcinoma (NPC). In this study, bioactivity-guided fractionation of the anti-NPC compound(s) from C. minima led to the isolation of 2beta (isobutyryloxy)florilenalin (IF), a sesquiterpene lactone. IF showed significant dose- and time- dependent inhibition on the growth of the human nasopharyngeal carcinoma epithelia cells (CNE). It induced apoptosis in CNE cells, as shown by the accumulation of sub-G1 cell population, DNA fragmentation and nuclear condensation, caspase-3 activation and PARP cleavage. Such induction was associated with the depletion of mitochondrial membrane potential (DeltaPsim) and the release of cytochrome c to cytosol to regulate the expression of Bcl-2 family proteins. These activities led to the cleavage of caspases and the trigger of cell death process. Overall, IF in C. minima showed potent antiproliferative effect of C. minima on NPC cells, suggesting that the plant deserves more extensive investigation for its potential medicinal application. PMID- 19553888 TI - Cyclization of hydrazones of 2-acetyl-1-naphthol and 1-acetyl-2-naphthol with triphosgene. Synthesis of Spiro naphthoxazine dimers. AB - Cyclization of hydrazones derived from 2-acetyl-1-naphthol and 1-acetyl-2 naphthol with triphosgene gave naphtho[1,2-e]-1,3-oxazines, naphtho[2,1-e]-1,3 oxazines or their spiro dimers depending on the molar ratio of triphosgene used for the cyclization. PMID- 19553889 TI - Extraction with SPME and synthesis of 2-methyl-6-vinylpyrazine by a 'one pot' reaction using microwaves. AB - A synthesis of 2-methyl-6-vinylpyrazine was carried out by way of a 'one pot' reaction. In order to establish the efficiency of this synthesis the extraction of the volatiles released by male papaya fruit flies was performed by SPME (solid phase micro-extraction). The compound was separated and identified using GC/MSD (gas chromatography/mass spectrometry detector). PMID- 19553890 TI - Effect of extraction solvent/technique on the antioxidant activity of selected medicinal plant extracts. AB - Theeffects of four extracting solvents [absolute ethanol, absolute methanol, aqueous ethanol (ethanol: water, 80:20 v/v) and aqueous methanol (methanol: water, 80:20 v/v)] and two extraction techniques (shaking and reflux) on the antioxidant activity of extracts of barks of Azadirachta indica, Acacia nilotica, Eugenia jambolana, Terminalia arjuna, leaves and roots of Moringa oleifera, fruit of Ficus religiosa,and leaves of Aloe barbadensis were investigated. The tested plant materials contained appreciable amounts of total phenolic contents (0.31 16.5 g GAE /100g DW), total flavonoid (2.63-8.66 g CE/100g DW); reducing power at 10 mg/mL extract concentration (1.36-2.91), DPPH(.) scavenging capacity (37.2 86.6%), and percent inhibition of linoleic acid (66.0-90.6%). Generally higher extract yields, phenolic contents and plant material antioxidant activity were obtained using aqueous organic solvents, as compared to the respective absolute organic solvents. Although higher extract yields were obtained by the refluxing extraction technique, in general higher amounts of total phenolic contents and better antioxidant activity were found in the extracts prepared using a shaker. PMID- 19553891 TI - Synthesis of two new hemisynthetic diterpenylhydroquinones from natural ent labdanes. AB - The synthesis and structural determination of two new diterpenylhydroquinones: 2beta-acetoxy-15-phenyl-(22,25-dihydroxy)-ent-labda-8(17),13(E)-diene(1) and 2beta-hydroxy-15-phenyl-(22,25-dihydroxy)-ent-labda-8(17),13(E)-dieneis reported (2). These compounds were obtained by coupling via Electrophilic Aromatic Substitution (EAS) of 1,4-hydroquinone with primary or tertiary allyl alcohol derivatives of the natural ent-labdanes 3 and 4. With this new method, the best results were observed when mixtures of the primary alcohol derivatives 5-6 (26% yield of compound 1) and diol derivatives 9-10 (28% yield of compound 2) were used. PMID- 19553892 TI - Determination of the volatile composition of Rhodobryum giganteum (Schwaegr.) Par. (Bryaceae) using solid-phase microextraction and gas chromatography/mass spectrometry (GC/MS). AB - A total of 38 volatile components were identified in Rhodobryum giganteum (Schwaegr.) Par. collected from two different geographic regions by headspace solid-phase microextraction (HS-SPME), combined with gas chromatography/mass spectrometry (GC/MS). The volatile components included some aliphatic and aromatic aldehydes, monoterpene hydrocarbons and a sesquiterpene (alpha farnesene), with 1-methoxy-2-propyl acetate and n-hexanal being found to be the most abundant volatile components. Analysis of the chemical constituents in the volatile oil of the two samples showed that ten compounds were shared. PMID- 19553894 TI - Synthesis and characterization of photo-responsive carbosilane dendrimers. AB - Preparation of photo-responsive carbosilane dendrimers bearing 4-phenylazo benzonitrile units on their molecular surface has been accomplished, and their both photo and thermal behaviors have also been characterized. These functional dendrimers suggest that the apparent molecular sizes of the cis-isomers are smaller than those of the corresponding trans-isomers, since the molecular diameter of these dendrimers would be shorter on the basis of trans-->cis photo isomerization of azobenzene. PMID- 19553893 TI - Examination of imprinting process with molsidomine as a template. AB - Eight different functional monomers were used with ethylene glycol dimethacrylate as a cross-linker and molsidomine as a template to obtain molecularly imprinted polymers (MIPs). Non-covalent interactions between molsidomine and each functional monomer in DMSO prior to thermal bulk polymerization were utilized. On the basis of calculated imprinting factors, MIP prepared with N,N' diallyltartaramide was chosen for further investigations. Examination of interactions in the prepolymerization complex between molsidomine and N,N' diallyltartaramide was performed using the Job method. The absorbance of isomolar solutions reaching a maximum for the molar ratio of template to monomer equal to 1:4. Scatchard analysis was used for estimation of the dissociation constants and the maximum amounts of binding sites. The polymer based on N,N' diallyltartaramide has two classes of heterogeneous binding sites characterized by two values of K(d) and two B(max): K(d)(1) = 1.17 mM(-1) and B(max)(1) = 0.8 mumol/mg for the higher affinity binding sites, and K(d)(2) = 200 microM(-1) and B(max)(2) = 2.05 mumol/mg for the lower affinity binding sites. Furthermore, effects of pH and organic solvent on binding properties of MIP and NIP were investigated, together with release of molsidomine from both MIP and NIP. PMID- 19553895 TI - A facile route to C2-substituted imidazolium ionic liquids. AB - A convenient route for the preparation of C2-substituted imidazolium ionic liquids is reported. This method involves the alkylation of N-heterocyclic carbenes, which are readily generated from the C2-unsubstituted imidazolium ionic liquids. It works well for non-functionalized alkyl chlorides, and less well for alkyl bromides and iodides, likely due to competing elimination reactions. The resulting C2-substituted salts can be transformed into ionic liquids via standard anion metathesis reactions. PMID- 19553896 TI - Two new steroidal saponins from Allium macrostemon bunge and their cytotoxity on different cancer cell lines. AB - Two new steroidal saponins (1 and 2) were isolated from the dried bulbs of Allium macrostemon Bunge. Their structures were elucidated by the spectral data as 26-O beta-D-glucopyranosyl-5alpha-furost-25 (27)-ene-3beta, 12beta, 22, 26-tetraol-3-O beta-D-glucopyranosyl (1-->2) [beta-D-glucopyranosyl (1-->3)]-beta-D glucopyranosyl (1-->4)-beta-D-galactopyranoside (1) and 26-O-beta-D glucopyranosyl-5beta-furost-20 (22)-25 (27)-dien-3beta, 12beta, 26-triol-3-O-beta D-glucopyranosyl (1-->2)-beta-D-galactopyranoside (2), respectively. Their cytotoxic activities on several cancer cell lines (MCF-7, NCI-H460, SF-268 and HepG2) were tested. 1 showed special cytotoxity on SF-268, while 2 showed cytotoxity on NCI-H460 and SF-268 cell lines, respectively. PMID- 19553897 TI - Heterocyclic-2-carboxylic acid (3-cyano-1,4-di-N-oxidequinoxalin-2-yl)amide derivatives as hits for the development of neglected disease drugs. AB - Neglected diseases represent a major health problem. It is estimated that one third of the world population is infected with tuberculosis (TB). Besides TB, Chagas disease, affects approximately 20 million people. Quinoxalines display great activities against TB and Chagas. Forty new quinoxaline 1,4-di-N-oxide derivatives have been prepared and tested against M. tuberculosis and T. cruzi. Carboxylic acid quinoxaline 1,4-di-N-oxides (CAQDOs) 5 and 17 showed MIC values on the same order as the reference antituberculosis drug, rifampicin. Meanwhile, CAQDOs 12 and 22 presented IC(50) values in the same order as the anti-chagasic drug, nifurtimox. PMID- 19553898 TI - A new cadinane sesquiterpene from the marine brown alga Dictyopteris divaricata. AB - A sample of the marine brown alga D. divaricata collected off the coast of Yantai (P.R. China) was dried, powdered, and extracted with the mixture of CHCl(3) and MeOH (1:1, v/v). By a combination of silica gel and Sephadex LH-20 column chromatography and preparative TLC, a new cadinane sesquiterpene 1,4 epoxymuurolan-5beta-ol (1) was isolated from this species. Its structure was established by detailed MS and NMR spectroscopic analysis, as well as comparison with literature data. PMID- 19553899 TI - Synthesis of symmetrical and non-symmetrical diimines from dimedone. AB - Symmetrical and non-symmetrical diimines derived from dimedone were synthesized by the reaction of their corresponding enaminothiones with primary amines. The synthesized compounds were characterized using micro analytical data and NMR spectroscopy. Theoretical calculations by B3LYP/6-31G(d,p) level of theory show that the enolic form is the most stable within the possible tautomeric forms of the compounds. PMID- 19553901 TI - Late HIV testing - 34 states, 1996-2005. AB - Without effective antiretroviral therapy, most persons infected with human immunodeficiency virus (HIV) will progress to acquired immunodeficiency syndrome (AIDS) in approximately 10 years (1). Testing, diagnosis, and medical care soon after HIV infection and before developing AIDS can prevent unnecessary morbidity and mortality and reduce further HIV transmission. Persons who receive an AIDS diagnosis concurrently or soon after receiving their initial HIV diagnosis (e.g., or=95%) vaccination coverage with 2 doses of measles-containing vaccine (MCV1 and MCV2) through routine vaccination and/or supplemental immunization activities (SIAs); 2) high-quality case-based measles surveillance; and 3) access to an accredited measles laboratory network for testing of suspected measles cases and identification of measles virus genotypes. This report describes progress toward measles elimination in the WPR through 2008. Measles likely has been eliminated or nearly eliminated in 24 of the 37 countries and areas in the WPR (referred to in this report as countries). However, large numbers of measles cases continue to be reported from several countries. During 2008, a total of 131,441 confirmed measles cases (98.4 per million population) were reported from China and 11,015 cases (86.1 per million population) from Japan, two countries that account for 82% of the region's population and >97% of its confirmed measles cases. Intensified efforts by WPR countries, particularly China and Japan, will be required to achieve the 2012 goal. PMID- 19553904 TI - Updated recommendations for use of Haemophilus influenzae type b (Hib) vaccine: reinstatement of the booster dose at ages 12-15 months. AB - On December 13, 2007, certain lots of Haemophilus influenzae type b (Hib) vaccine marketed as PedvaxHIB (monovalent Hib vaccine) and Comvax (Hib-HepB vaccine), and manufactured by Merck & Co., Inc., were recalled voluntarily, and the company temporarily suspended production of these vaccines. To conserve the limited supply of Hib-containing vaccines, CDC, in consultation with the Advisory Committee on Immunization Practices (ACIP), the American Academy of Family Physicians (AAFP), and the American Academy of Pediatrics (AAP), on December 18, 2007, recommended that vaccination providers temporarily defer the routine Hib vaccine booster dose administered to most healthy children at age 12-15 months. PMID- 19553905 TI - Implantation of engineered tissue in the rat heart. AB - Rodent surgery is often an important component in assessing the utility of engineered tissues. A wide variety of surgical procedures can be performed in common laboratory rats or mice and these quite frequently serve as an intermediate step between bench-top experiments and large animal testing or human trials. Given that rodents provide an established, cost-effective, and physiologically-relevant model system in which to test novel combinations of scaffolding materials and cells, they are particularly well-suited for cardiovascular tissue engineering studies. Presently, we describe an open-heart surgical procedure to implant engineered tissue containing myogenic progenitor cells in the atrioventricular (AV) groove of a rat heart. These implants are intended to create an electrical conduit between the right atrium and right ventricle with the ultimate goal of providing an alternative treatment to conventional pacemaker implantation in pediatric patients with complete heart block. The engineered tissue is implanted in the AV-groove by means of a thoracotomy. For our purposes, Lewis rats are anesthetized and invasively ventilated to maintain positive airway pressure during the sterile surgical procedure. The approach to the heart is performed by a right thoracotomy through an antero-lateral incision at the 5(th) intercostal space. The tissue construct is fixed in the AV groove using a single 7-0 Prolene suture and positioned between the right ventricle and atrium at the ventral portion of the heart. The epicardium is partially removed to allow direct contact between the recipient myocardial cells and those contained in the engineered tissue. Following implantation, the chest wall is closed in layers, any pneumothorax is evacuated, and the animal is extubated and treated with analgesic. PMID- 19553906 TI - Rosiglitazone alone or in combination with tissue plasminogen activator improves ischemic brain injury in an embolic model in rats. AB - In this study, we examined whether rosiglitazone, a peroxisome proliferator activated receptor gamma (PPARgamma) agonist, is neuroprotective in focal ischemic brain injury, and whether rosiglitazone can enhance the protective action of tissue plasminogen activator (tPA), an agent used clinically for thrombolytic therapy. Rats were subjected to ischemic brain injury by embolizing preformed clots into the middle cerebral artery (MCA). Treatment with rosiglitazone reduced infarction and improved functional recovery; it also enhanced the neuroprotective action of tPA and lengthened the time window for initiating tPA treatment. Occlusion of MCA resulted in a loss of collagen type IV, a major structural protein of the microvascular basal lamina, and tPA treatment worsened this loss. Rosiglitazone treatment prevented the reduction of collagen type IV in the ischemic injured brain by inhibiting the activation of matrix metallopeptidase-9 (MMP-9). In addition, rosiglitazone treatment reduced inflammatory reactions in the ischemic injured brain. Rosiglitazone either alone or in combination with tPA is an effective agent in the reduction of ischemic brain injury. The reduction of microvascular damage and inflammation contributes to the beneficial actions of rosiglitazone. PMID- 19553907 TI - The protective effect of early hypothermia on PTEN phosphorylation correlates with free radical inhibition in rat stroke. AB - We recently showed that intraischemic moderate hypothermia (30 degrees C) reduces ischemic damage through the Akt pathway after permanent distal middle cerebral artery occlusion in rats. The only Akt pathway component preserved by hypothermia is phosphorylated phosphatase and tensin homolog deleted on chromosome 10 (p PTEN), which suggests that p-PTEN may have a central role in neuroprotection. Reactive oxygen species (ROS) are critically involved in mediating ischemic damage after stroke by interacting with signaling molecules, including Akt, PTEN, and delta-protein kinase C (PKC). We investigated the protective mechanisms of moderate hypothermia on these signaling proteins after transient focal ischemia in rats. Early moderate hypothermia (3 h) was administered 15 mins before reperfusion, and delayed moderate hypothermia (3 h) was applied 15 mins after reperfusion. Our results indicate that early hypothermia reduced infarction, whereas delayed hypothermia did not. However, both early and delayed hypothermia maintained levels of Mn-SOD (superoxide dismutase) and phosphorylated Akt and blocked delta-PKC cleavage, suggesting that these factors may not be critical to the protection of hypothermia. Nevertheless, early hypothermia preserved p-PTEN levels after reperfusion, whereas delayed hypothermia did not. Furthermore, ROS inhibition maintained levels of p-PTEN after stroke. Together, these findings suggest that phosphorylation levels of PTEN are closely associated with the protective effect of early hypothermia against stroke. PMID- 19553908 TI - Cardiac arrest and cardiopulmonary resuscitation dysregulates the hypothalamic pituitary-adrenal axis. AB - Cardiac arrest and cardiopulmonary resuscitation (CA/CPR) increase the risk for affective disorders in human survivors. Postischemic anxiety- and depressive-like behaviors have been documented in animal models of CA/CPR; however, the stability of post-CA/CPR anxiety-like behavior over time and the underlying physiologic mechanisms remain unknown. The hypothalamic-pituitary-adrenal (HPA) axis and the corticotropin releasing factor (CRF) system may mediate the pathophysiology of anxiety and depression; therefore, this study measured CA/CPR-induced changes in CRF receptor binding and HPA axis negative feedback. Mice were exposed to CA/CPR or SHAM surgery and assessed 7 or 21 days later. Consistent with earlier demonstrations of anxiety-like behavior 7 days after CA/CPR, increased anxiety like behavior in the open field was also present 21 days after CA/CPR. On postoperative day 7, CA/CPR was associated with an increase in basal serum corticosterone concentration relative to SHAM, but this difference resolved by postoperative day 21. The Dexamethasone Suppression Test showed that the CA/CPR group had enhanced negative feedback compared with SHAM controls at postoperative day 21. Furthermore, there was a gradual increase in CRF(1) receptor binding in the paraventricular nucleus of the hypothalamus and bed nucleus of the stria terminalis, as well as a transient decrease of both CRF(1) and CRF(2A) receptors in the dorsal hippocampus. Therefore, sustained changes in activity of the HPA axis and the CRF system after CA/CPR may contribute to the postischemic increase in affective disorders. PMID- 19553909 TI - Noninvasive assessment of the brain redox status after transient middle cerebral artery occlusion using Overhauser-enhanced magnetic resonance imaging. AB - Oxidative stress has been implicated in the cell death that occurs after ischemia reperfusion of the brain, which causes the production of reactive oxygen species and a decrease in antioxidants, leading to mitochondrial dysfunction. However, the invasive methods used to collect much of this evidence are themselves stress inducing, which could skew the results. In this study, we aimed at demonstrating brain redox alterations after ischemia-reperfusion noninvasively, using Overhauser-enhanced magnetic resonance imaging. The reduction rate of 3 methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-L-oxyl (methoxycarbonyl-PROXYL), a redox-sensitive contrast agent, was used as an index of the redox status in vivo. No changes were observed in the antioxidant concentration, the mitochondrial complex activity, or in the redox status image intensity after 3 h of reperfusion, following transient middle cerebral artery occlusion; however, after 24 h of reperfusion, the methoxycarbonyl-PROXYL reduction rate, calculated from continuous images, had decreased significantly. Concordantly, biochemical assays showed that the concentration of ascorbic acid in the ischemic hemisphere and the activity of mitochondrial complex II had also decreased. Thus, the noninvasive imaging of the brain redox alterations faithfully reflected changes in antioxidant levels and in mitochondrial complex II activity after ischemia reperfusion. PMID- 19553910 TI - Indigenous people in Australia, Canada, New Zealand and the United States are less likely to receive renal transplantation. AB - In Australia, Canada, New Zealand, and the United States indigenous people have high rates of chronic kidney disease but poor access to effective therapies. To more fully define these issues, we compared the demographics of renal transplantation of indigenous patients in these 4 countries. Data encompassing 312,507 indigenous and white patients (18-64 years of age) who initiated dialysis within an 11-year period ending in 2005 were obtained from each country's end stage kidney disease registry. By the study's end, 88,173 patients had received a renal transplant and 130,261 had died without receiving such. Compared with white patients, the adjusted likelihood of receiving a transplant for indigenous patients was significantly lower in Australia (hazard ratio (HR) 0.23), Canada (HR 0.34), New Zealand (HR 0.23), and the United States (HR 0.44). In all four countries, indigenous patients had significantly longer overall median waiting times compared to white patients. Our study shows that despite marked differences in health care delivery systems, indigenous patients are less likely than white patients to receive a renal transplant in these countries. Understanding and addressing barriers to renal transplantation of indigenous patients remains an important concern. PMID- 19553911 TI - Deletion of LOX-1 attenuates renal injury following angiotensin II infusion. AB - Angiotensin II upregulates the expression of LOX-1, a recently identified oxidized low-density lipoprotein receptor controlled by redox state which in turn upregulates angiotensin II activity on its activation. To test whether interruption of this positive feedback loop might reduce angiotensin II-induced hypertension and subsequent renal injury, we studied LOX-1 knockout mice. After infusion with angiotensin II for 4 weeks systolic blood pressure gradually increased in the wild-type mice; this rise was significantly attenuated in the LOX-1 knockout mice. Along with the rise in systolic blood pressure, renal function (blood urea nitrogen and creatinine) decreased in the wild-type mice, but the deterioration of function was significantly less in the LOX-1 knockout mice. Glomerulosclerosis, arteriolar sclerosis, tubulointerstitial damage, and renal collagen accumulation were all significantly less in the LOX-1 knockout mice. The reduction in collagen formation was accompanied by a decrease in connective tissue growth factor mRNA, angiotensin type 1 receptor expression, and phosphorylation of p38 and p44/42 mitogen-activated protein kinases. Expression of endothelial nitric oxide synthase was increased in the kidneys of the LOX-1 knockout mice compared to the wild-type mice. Overall, our study suggests that LOX-1 is a key modulator in the development of angiotensin II-induced hypertension and subsequent renal damage. PMID- 19553912 TI - Posttransplant anemia: the role of sirolimus. AB - Posttransplant anemia is a common problem that may hinder patients' quality of life. It occurs in 12 to 76% of patients, and is most common in the immediate posttransplant period. A variety of factors have been identified that increase the risk of posttransplant anemia, of which the level of renal function is most important. Sirolimus, a mammalian target of rapamycin inhibitor, has been implicated as playing a special role in posttransplant anemia. This review considers anemia associated with sirolimus, including its presentation, mechanisms, and management. PMID- 19553913 TI - Transglutaminase inhibition ameliorates experimental diabetic nephropathy. AB - Diabetic nephropathy is characterized by excessive extracellular matrix accumulation resulting in renal scarring and end-stage renal disease. Previous studies have suggested that transglutaminase type 2, by formation of its protein crosslink product epsilon-(gamma-glutamyl)lysine, alters extracellular matrix homeostasis, causing basement membrane thickening and expansion of the mesangium and interstitium. To determine whether transglutaminase inhibition can slow the progression of chronic experimental diabetic nephropathy over an extended treatment period, the inhibitor NTU281 was given to uninephrectomized streptozotocin-induced diabetic rats for up to 8 months. Effective transglutaminase inhibition significantly reversed the increased serum creatinine and albuminuria in the diabetic rats. These improvements were accompanied by a fivefold decrease in glomerulosclerosis and a sixfold reduction in tubulointerstitial scarring. This was associated with reductions in collagen IV accumulation by 4 months, along with reductions in collagens I and III by 8 months. This inhibition also decreased the number of myofibroblasts, suggesting that tissue transglutaminase may play a role in myofibroblast transformation. Our study suggests that transglutaminase inhibition ameliorates the progression of experimental diabetic nephropathy and can be considered for clinical application. PMID- 19553914 TI - Susceptibility locus in neurokinin-1 receptor gene associated with alcohol dependence. AB - Substance P (SP), a neurotransmitter in stress pathways, exerts its effects mainly through the neurokinin-1 receptor (NK1R). Genetic and pharmacological studies show that binding of ligands to NK1R decreases anxiety-related behaviors, and therefore, self-administration of alcohol in mice and craving for alcohol in humans. As genetic variants may result in differential expression of the receptor through various molecular mechanisms, we examined whether allelic variations in the NK1R gene are associated with alcohol dependence (AD) by genotyping 11 single nucleotide polymorphisms (SNPs) across NK1R in alcoholic (n=271) and healthy control (n=337) participants of Caucasian descent. The AD was diagnosed using the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, fourth edition. Associations of the SNPs with AD were assessed at both the individual SNP and haplotype levels. We found that genotype and allele frequencies of rs6715729, a synonymous SNP in exon 1, differed significantly in alcoholics and in controls (p=0.0006; OR (odds ratio)=6.13; 95% CI=4.06, 9.23). Haplotype analyses indicated two risk haplotypes for AD in the 5' end of the gene, formed by the three-SNP combinations rs6715729-rs735668-rs6741029. Taken together, we conclude that polymorphisms of NK1R are significantly associated with the development of AD in Caucasian individuals. Additional studies are needed to replicate these results in other samples and to elucidate the mechanism(s) by which these polymorphisms affect NK1R function in the brain. PMID- 19553915 TI - Adolescent exposure to chronic delta-9-tetrahydrocannabinol blocks opiate dependence in maternally deprived rats. AB - Maternal deprivation in rats specifically leads to a vulnerability to opiate dependence. However, the impact of cannabis exposure during adolescence on this opiate vulnerability has not been investigated. Chronic dronabinol (natural delta 9 tetrahydrocannabinol, THC) exposure during postnatal days 35-49 was made in maternal deprived (D) or non-deprived (animal facility rearing, AFR) rats. The effects of dronabinol exposure were studied after 2 weeks of washout on the rewarding effects of morphine measured in the place preference and oral self administration tests. The preproenkephalin (PPE) mRNA levels and the relative density and functionality of CB1, and mu-opioid receptors were quantified in the striatum and the mesencephalon. Chronic dronabinol exposure in AFR rats induced an increase in sensitivity to morphine conditioning in the place preference paradigm together with a decrease of PPE mRNA levels in the nucleus accumbens and the caudate-putamen nucleus, without any modification for preference to oral morphine consumption. In contrast, dronabinol treatment on D-rats normalized PPE decrease in the striatum, morphine consumption, and suppressed sensitivity to morphine conditioning. CB1 and mu-opioid receptor density and functionality were not changed in the striatum and mesencephalon of all groups of rats. These results indicate THC potency to act as a homeostatic modifier that would worsen the reward effects of morphine on naive animals, but ameliorate the deficits in maternally D-rats. These findings point to the self-medication use of cannabis in subgroups of individuals subjected to adverse postnatal environment. PMID- 19553917 TI - Impaired error awareness and anterior cingulate cortex hypoactivity in chronic cannabis users. AB - Drug abuse and other psychiatric conditions (eg, schizophrenia) have been associated with a diminished neural response to errors, particularly in the anterior cingulate cortex (ACC) thought critical to error processing. A diminished capacity for detecting errors has been linked to clinical symptoms including the loss of insight, delusions, and perseverative behavior. A total of 16 active chronic cannabis users and 16 control participants were administered a Go/No-go response inhibition task during event-related fMRI data collection. The task provides measures of inhibitory control and error awareness. Cannabis users' inhibitory control performance was equivalent to that of the control group, but the former showed a significant deficit in awareness of commission errors. Cannabis users showed a diminished capacity for monitoring their behavior that was associated with hypoactivity in the ACC and right insula. In addition, increased levels of hypoactivity in both the ACC and right insula regions were significantly correlated with error-awareness rates in the cannabis group (but not controls). These difficulties are consistent with earlier reports of hypoactivity in the neural systems underlying cognitive control and the monitoring of interoceptive awareness in chronic drug users, and highlight the potential relationship between cognitive dysfunction and behavioral deficits that have the potential to contribute to the maintenance of drug abuse. PMID- 19553916 TI - Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment: pharmacological and cellular characterization. AB - This study extends earlier work on the role of vascular endothelial growth factor (VEGF) in the actions of antidepressant treatment in two key areas. First, by determining the requirement for VEGF in the actions of a 5-HT selective reuptake inhibitor (SSRI), fluoxetine in behavioral models of depression/antidepressant response; and second, by examining the role of the 5-HT1A receptor subtype in the regulation of VEGF, and the cellular localization of antidepressant regulation of VEGF expression. The results show that pharmacological inhibition of VEGF receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress. Infusions of SU5416 or SU1498, two structurally dissimilar inhibitors of VEGF-Flk-1 receptor signaling, block the antidepressant effects of fluoxetine on sucrose preference, immobility in the forced swim test, and latency to feed in the novelty suppressed feeding paradigm. We also show that activation of 5-HT1A receptors is sufficient to induce VEGF expression and that a 5-HT1A antagonist blocks both the increase in VEGF and behavioral effects induced by fluoxetine. Finally, double labeling studies show that chronic fluoxetine administration increases VEGF expression in both neurons and endothelial cells in the hippocampus. Taken together these studies show that VEGF is necessary for the behavioral effects of the SSRI fluoxetine, as well as norepinephrine selective reuptake inhibitor, and that these effects may be mediated by 5-HT1A receptors located on neurons and endothelial cells. PMID- 19553920 TI - Massively obese adolescents were of normal weight at the age of adiposity rebound. PMID- 19553918 TI - Alcohol exposure alters NMDAR function in the bed nucleus of the stria terminalis. AB - Chronic alcohol exposure can cause dramatic behavioral alterations, including increased anxiety-like behavior and depression. These alterations are proposed to be due in part to adaptations in the brain regions that regulate emotional behavior, including the bed nucleus of the stria terminalis (BNST), a principal output nucleus of the amygdala. However, to date there have been no studies that have examined the impact of in vivo alcohol exposure on synaptic function in the BNST. To better understand how alcohol can alter neuronal function, we examined the ability of in vivo alcohol exposure to alter glutamatergic transmission in the BNST using whole-cell voltage clamp recordings and biochemistry in brain slices obtained from C57Bl6 mice. Chronic intermittent, but not continuous, ethanol vapor exposure increased temporal summation of NMDA receptor (NMDAR) mediated excitatory postsynaptic currents (EPSCs). Both electrophysiological and biochemical approaches suggest that this difference is not because of an alteration in glutamate release, but rather an increase in the levels of NR2B containing NMDARs. Further, we found that ethanol modulation of NMDAR in the vBNST is altered after intermittent alcohol exposure. Our results support the hypothesis that NMDAR-mediated synaptic transmission is sensitized at key synapses in the extended amygdala and thus may be a suitable target for manipulation of the behavioral deficits associated with acute withdrawal from chronic alcohol exposure. PMID- 19553921 TI - High-fat diets: details matter. PMID- 19553922 TI - Early adiposity rebound is an important predictor of later obesity. PMID- 19553923 TI - A complementary explanation on the relationship of obesity and oxidative stress via protons. PMID- 19553924 TI - Effects of chronic treatment with the CB1 antagonist, rimonabant on the blood pressure, and vascular reactivity of obese Zucker rats. AB - Rimonabant (RM) is a cannabinoid CB1 receptor antagonist useful in the treatment of obesity associated cardiovascular risk factors. Since cannabinoids are vasoactive compounds, the aim of this study is to evaluate the effect of chronic treatment with RM on systolic blood pressure (SBP), and endothelial and vascular reactivity. Obese Zucker rats (OZRs) and their lean counterparts were orally treated during 20 weeks with either RM (10 mg/kg/day). Endothelial and vascular function was assessed in aorta and small mesenteric arteries (SMAs) by concentration response curves to acetylcholine (ACh) and phenylephrine (Phe), respectively. Participation of nitric oxide (NO) was evaluated by incubation with the NO synthase (NOS) inhibitor N(G)-nitro-l-arginine methyl ester (L-NAME) and cyclooxygenase (COX)-derived products involvement was analyzed by incubation with indomethacin (INDO). Plasma lipid profile, insulin and adiponectin were also analyzed. Sympathetic activity was evaluated by urinary excretion of noradrenaline. As expected, RM decreased body weight gain and enhanced adiponectin concentration. Insulin resistance and sympathetic activity were also decreased. The increase in SBP observed in OZRs was reduced by treatment with RM. Aortae and SMAs from OZRs exhibited lower contractile response to Phe, being this effect prevented by RM administration. Although ACh-induced response and NO participation remained unaltered with obesity, enhanced COX-derived constrictor products were found in OZRs. RM treatment neither altered endothelium-dependent relaxation nor L-NAME-sensitive component of the response. Nevertheless, it was able to regulate COX-derived vasoactive products participation. Those effects may contribute to explain some of the cardiovascular protective actions elicited by this drug. PMID- 19553925 TI - Nonalcoholic hepatic steatosis in Zucker diabetic rats: spontaneous evolution and effects of metformin and fenofibrate. AB - No specific treatment for nonalcoholic hepatic fatty liver disease has been defined. We followed the spontaneous evolution of liver steatosis and tested the therapeutic usefulness of metformin and fenofibrate in a model of steatosis, the Zucker diabetic fatty (ZDF) rat. ZDF and control rats were studied at 7, 14, and 21 weeks. After initial study at 7 weeks, ZDF rats received no treatment, metformin or fenofibrate until studies at 14 or 21 weeks. ZDF rats were obese, hypertriglyceridemic, insulin resistant at 7 weeks, type 2 diabetic at 14, diabetic with insulin deficiency at 21. They had steatosis at 7 weeks with increased hepatic expression and activity of lipogenesis. Steatosis was unchanged at 14 and 21 weeks despite lower expression and activity of lipogenesis. Metformin and fenofibrate did not modify energy intake or expenditure or the evolution of diabetes. Both compounds decreased plasma triacylglycerol (TAG) concentrations. Hepatic TAG content was reduced by fenofibrate at 14 and 21 weeks but only at 21 weeks by metformin. Metformin had no significant effects on the expression in liver of genes of fatty acids metabolism. The beneficial effect of fenofibrate occurred despite increased expression of genes involved in the uptake and activation of fatty acids. Acyl-CoA oxidase (ACO) and carnitine palmitoyltransferase I (CPTI) mRNA levels were increased by fenofibrate showing evidence of increased lipid oxidation. To conclude, metformin had only moderate effects on liver steatosis. The effects of fenofibrate was more marked but remained mild. PMID- 19553926 TI - Association of the CPT1B gene with skeletal muscle fat infiltration in Afro Caribbean men. AB - Skeletal muscle fat is greater in African ancestry individuals compared with whites, is associated with diabetes, and is a heritable polygenic trait. However, specific genetic factors contributing to skeletal muscle fat in humans remain to be defined. Muscle carnitine palmitoyltransferase-1B (CPT1B) is a key enzyme in the regulation of skeletal muscle mitochondrial beta-oxidation of long-chain fatty acids, and as such is a reasonable biological candidate gene for skeletal muscle fat accumulation. Therefore, we examined the association of three nonsynonymous coding variants in CPT1B (G531L, I66V, and S427C; a fourth, A320G, could not be genotyped) and quantitative computed tomography measured tibia skeletal muscle composition and BMI among 1,774 Afro-Caribbean men aged > or =40, participants of the population-based Tobago Health Study. For all variants, no significant differences were observed for BMI or total adipose tissue. Among individuals who were homozygous for the minor allele at G531L or I66V, intermuscular adipose tissue (IMAT) was 87% (P = 0.03) and 54% lower (P = 0.03), respectively. In contrast, subcutaneous adipose tissue (SAT) was 11% (P = 0.017) and 7% (P = 0.049) higher, respectively, than among individuals without these genotypes. These associations were independent of age, body size, and muscle area. Finally, no individuals with type 2 diabetes were found among those who were homozygous for the minor allele of either at G531L and I66V whereas 14-18% of men with the major alleles had type 2 diabetes (P = 0.03 and 0.007, respectively). Our results suggest a novel association between common nonsynonymous coding variants in CPT1B and ectopic skeletal muscle fat among middle-aged and older African ancestry men. PMID- 19553927 TI - Optimal body weight for the prevention of coronary heart disease in normal-weight physically active men. AB - Although 36% of US men are normal weight (BMI <25 kg/m(2)), the health benefits of greater leanness in normal-weight individuals are seldom acknowledged. To assess the optimal body weight with respect to minimizing coronary heart disease (CHD) risk, we applied Cox proportional hazard analyses of 20,525 nonsmoking, nondiabetic, normal-weight men followed prospectively for 7.7 years, including 20,301 who provided follow-up questionnaires. Two-hundred and forty two men reported coronary artery bypass graph (CABG) or percutaneous transluminal coronary angioplasty (PTCA) and 82 reported physician-diagnosed incident myocardial infarction (267 total). The National Death Index identified 40 additional ischemic heart disease deaths. In these normal-weight men, each kg/m(2) decrement in baseline BMI was associated with 11.2% lower risk for total CHD (P = 0.005), 13.2% lower risk for nonfatal CHD (P = 0.002), 19.0% lower risk for nonfatal myocardial infarction (P = 0.01), and 12.2% lower risk for PTCA or CABG (P = 0.007). Compared to men with BMI between 22.5 and 25 kg/m(2), those <22.5 kg/m(2) had 24.1% lower total CHD risk (P = 0.01), 27.9% lower nonfatal CHD risk (P = 0.01), 37.8% lower nonfatal myocardial infarction risk (P = 0.05), and 27.8% lower PTCA or CABG risk (P = 0.02). In nonabdominally obese men (waist circumference <102 cm), CHD risk declined linearly with declining waist circumference. CHD risk was unrelated to change in waist circumference between 18 years old and baseline except as it contributed to baseline circumference. These results suggest that the optimal BMI for minimizing CHD risk lies somewhere <22.5 kg/m(2), as suggested from our previous analyses of incident diabetes, hypertension, and hypercholesterolemia in these men. PMID- 19553928 TI - Sustained NPY overexpression in the PVN results in obesity via temporarily increasing food intake. AB - Increasing neuropeptide Y (NPY) signaling in the paraventricular nucleus (PVN) by recombinant adeno-associated virus (rAAV)-mediated overexpression of NPY in rats, results in hyperphagia and obesity in rats. To determine the importance of hyperphagia in the observed obesity phenotype, we pair-fed a group of AAV-NPY injected rats to AAV-control-injected rats and compared parameters of energy balance to ad libitum fed AAV-NPY-injected rats. For 3 weeks, AAV-NPY-injected rats, received the same amount of food as ad libitum-fed rats injected with control rAAV They did not gain more body weight than these controls. When allowed access to food ad libitum, these AAV-NPY-injected rats increased food intake, which subsequently decreased when rats reached the same body weight as AAV-NPY injected rats that were fed ad libitum for the entire study. These data indicate that overexpression of NPY in the PVN results in obesity by increasing food intake until a certain body weight is achieved. PMID- 19553931 TI - Utility of adiponectin as a biomarker predictive of glycemic efficacy is demonstrated by collaborative pooling of data from clinical trials conducted by multiple sponsors. AB - This study, conducted under the Metabolic Disorders Steering Committee of the Biomarkers Consortium (a public-private partnership managed by the Foundation for the National Institutes of Health (FNIH)), analyzed blinded data on 2,688 type 2 diabetes (T2D) patients from randomized clinical trials conducted by four pharmaceutical companies. An increase in the levels of adiponectin was observed after peroxisome proliferator-activated receptor (PPAR)-agonist treatment (P < 0.0001), but not after treatment with non-PPAR drugs. This increase correlated with decreases in levels of glucose, hemoglobin A(1c) (Hb(A1c)), hematocrit, and triglycerides, and increases in levels of blood urea nitrogen, creatinine, and high-density lipoprotein cholesterol (HDL-C). Early (6-8 weeks) increases in levels of adiponectin after treatment with PPAR agonists showed a negative correlation (r = -0.21, P < 0.0001) with subsequent changes in levels of Hb(A1c). Changes in adiponectin level did not appear to be associated with baseline level of Hb(A1c). Logistic regression demonstrated that an increase in the level of adiponectin predicts a decrease in the level of Hb(A1c). These analyses confirm previously demonstrated relationships between adiponectin levels and metabolic parameters and support the robust predictive utility of adiponectin across the spectrum of glucose tolerance. Cross-company precompetitive collaboration is a feasible and powerful approach to biomarker qualification. PMID- 19553932 TI - New antithrombotic drugs. AB - Thrombosis, both venous and arterial, is a major cause of morbidity and mortality worldwide. Consequently, there is an ongoing search for new antithrombotic drugs, particularly novel antiplatelet agents and anticoagulants. A better understanding of the biochemical pathways involved in platelet activation and coagulation and of the links between these systems and the impact of thrombosis on inflammation has led to the identification of new targets for antithrombotic drugs. This paper focuses on these new targets and new antiplatelet drugs and anticoagulants and describes the major advances in the continuing search for more potent antithrombotic drugs that have limited effects on hemostasis. PMID- 19553933 TI - Pharmacological treatment for heart failure: a view from the brain. AB - Systolic heart failure is a feed-forward phenomenon with devastating consequences. Impaired cardiac function is the initiating event, but central nervous system mechanisms activated by persistent altered neural and humoral signals from the periphery play an important sustaining role. Animals with experimentally induced heart failure have neurochemical abnormalities in the brain that, when manipulated, profoundly affect sympathetic drive, volume regulation, and cardiac remodeling--critical determinants of outcome. This brief review explores recent studies that provide a strong rationale for the development of pharmaceutical agents that target central nervous system abnormalities in heart failure. PMID- 19553934 TI - COX-1 and vascular disease. AB - Since the discovery of the pivotal role of cyclooxygenase (COX) in the metabolism of arachidonic acid, vascular biologists have been confronted with the duality of this system. Indeed, one substrate (arachidonic acid) transformed by one enzyme (COX) yields end products (endoperoxides) that exist only very briefly before being metabolized to more stable prostanoids by a set of specific downstream synthases that were initially believed to be tissue specific. For instance, platelets contain mainly the synthase that produces thromboxane A(2) (a potent proaggregatory and vasoconstrictor substance), whereas endothelial cells contain mainly the enzyme that generates prostacyclin (an equally potent antiaggregatory and vasodilator substance). The overproduction of thromboxane A(2) by platelets leads to thrombosis; endothelial cells resist vascular occlusion by producing prostacyclin. This duality of the metabolism of arachidonic acid has dominated our thinking about atherothrombosis for decades, and rightfully still does. As scientific understanding progressed, it became evident that two isoforms of COX exist: COX-1 and COX-2. COX-1 was initially considered to be the "good," constitutive isoform, whereas COX-2 appeared to be mainly a "bad" inducible enzyme involved in inflammatory responses. However, more recently, the unexpected events resulting from the widespread use of selective COX-2 inhibitors has suggested that, from a cardiovascular point of view, the products of COX-2 exert a protective role and that this isoform cannot necessarily be regarded as "bad." Likewise, evidence has emerged that initiation of the metabolism of arachidonic acid by COX-1 is not necessarily a "good" thing in terms of vascular protection. This brief review focuses on the potential contribution of endothelial COX-1 to vascular dysfunction. It is based on a number of review articles, to which the reader will be referred in order to identify the original references to the statements made; these references are not cited here because of space limitations. PMID- 19553935 TI - New flutes document the earliest musical tradition in southwestern Germany. AB - Considerable debate surrounds claims for early evidence of music in the archaeological record. Researchers universally accept the existence of complex musical instruments as an indication of fully modern behaviour and advanced symbolic communication but, owing to the scarcity of finds, the archaeological record of the evolution and spread of music remains incomplete. Although arguments have been made for Neanderthal musical traditions and the presence of musical instruments in Middle Palaeolithic assemblages, concrete evidence to support these claims is lacking. Here we report the discovery of bone and ivory flutes from the early Aurignacian period of southwestern Germany. These finds demonstrate the presence of a well-established musical tradition at the time when modern humans colonized Europe, more than 35,000 calendar years ago. Other than the caves of the Swabian Jura, the earliest secure archaeological evidence for music comes from sites in France and Austria and post-date 30,000 years ago. PMID- 19553942 TI - Cheerleader or watchdog? PMID- 19553937 TI - A conserved ubiquitination pathway determines longevity in response to diet restriction. AB - Dietary restriction extends longevity in diverse species, suggesting that there is a conserved mechanism for nutrient regulation and prosurvival responses. Here we show a role for the HECT (homologous to E6AP carboxy terminus) E3 ubiquitin ligase WWP-1 as a positive regulator of lifespan in Caenorhabditis elegans in response to dietary restriction. We find that overexpression of wwp-1 in worms extends lifespan by up to 20% under conditions of ad libitum feeding. This extension is dependent on the FOXA transcription factor pha-4, and independent of the FOXO transcription factor daf-16. Reduction of wwp-1 completely suppresses the extended longevity of diet-restricted animals. However, the loss of wwp-1 does not affect the long lifespan of animals with compromised mitochondrial function or reduced insulin/IGF-1 signalling. Overexpression of a mutant form of WWP-1 lacking catalytic activity suppresses the increased lifespan of diet restricted animals, indicating that WWP-1 ubiquitin ligase activity is essential for longevity. Furthermore, we find that the E2 ubiquitin conjugating enzyme, UBC 18, is essential and specific for diet-restriction-induced longevity. UBC-18 interacts with WWP-1 and is required for the ubiquitin ligase activity of WWP-1 and the extended longevity of worms overexpressing wwp-1. Taken together, our results indicate that WWP-1 and UBC-18 function to ubiquitinate substrates that regulate diet-restriction-induced longevity. PMID- 19553943 TI - Raising the standards. PMID- 19553938 TI - A reevaluation of X-irradiation-induced phocomelia and proximodistal limb patterning. AB - Phocomelia is a devastating, rare congenital limb malformation in which the long bones are shorter than normal, with the upper portion of the limb being most severely affected. In extreme cases, the hands or fingers are attached directly to the shoulder and the most proximal elements (those closest to the shoulder) are entirely missing. This disorder, previously known in both autosomal recessive and sporadic forms, showed a marked increase in incidence in the early 1960s due to the tragic toxicological effects of the drug thalidomide, which had been prescribed as a mild sedative. This human birth defect is mimicked in developing chick limb buds exposed to X-irradiation. Both X-irradiation and thalidomide induced phocomelia have been interpreted as patterning defects in the context of the progress zone model, which states that a cell's proximodistal identity is determined by the length of time spent in a distal limb region termed the 'progress zone'. Indeed, studies of X-irradiation-induced phocomelia have served as one of the two major experimental lines of evidence supporting the validity of the progress zone model. Here, using a combination of molecular analysis and lineage tracing in chick, we show that X-irradiation-induced phocomelia is fundamentally not a patterning defect, but rather results from a time-dependent loss of skeletal progenitors. Because skeletal condensation proceeds from the shoulder to fingers (in a proximal to distal direction), the proximal elements are differentially affected in limb buds exposed to radiation at early stages. This conclusion changes the framework for considering the effect of thalidomide and other forms of phocomelia, suggesting the possibility that the aetiology lies not in a defect in the patterning process, but rather in progenitor cell survival and differentiation. Moreover, molecular evidence that proximodistal patterning is unaffected after X-irradiation does not support the predictions of the progress zone model. PMID- 19553944 TI - Orphan giant. PMID- 19553953 TI - Journal club. A physicist peels back the layers of excitement about graphene. PMID- 19553956 TI - New protein structures replace the old. PMID- 19553957 TI - US human spacefaring questioned. PMID- 19553958 TI - Deficit dooms Swedish gene institute. PMID- 19553959 TI - G8 cancels science parley. PMID- 19553936 TI - Evolution of a malaria resistance gene in wild primates. AB - The ecology, behaviour and genetics of our closest living relatives, the nonhuman primates, should help us to understand the evolution of our own lineage. Although a large amount of data has been amassed on primate ecology and behaviour, much less is known about the functional and evolutionary genetic aspects of primate biology, especially in wild primates. As a result, even in well-studied populations in which nongenetic factors that influence adaptively important characteristics have been identified, we have almost no understanding of the underlying genetic basis for such traits. Here, we report on the functional consequences of genetic variation at the malaria-related FY (DARC) gene in a well studied population of yellow baboons (Papio cynocephalus) living in Amboseli National Park in Kenya. FY codes for a chemokine receptor normally expressed on the erythrocyte surface that is the known entry point for the malarial parasite Plasmodium vivax. We identified variation in the cis-regulatory region of the baboon FY gene that was associated with phenotypic variation in susceptibility to Hepatocystis, a malaria-like pathogen that is common in baboons. Genetic variation in this region also influenced gene expression in vivo in wild individuals, a result we confirmed using in vitro reporter gene assays. The patterns of genetic variation in and around this locus were also suggestive of non-neutral evolution, raising the possibility that the evolution of the FY cis regulatory region in baboons has exhibited both mechanistic and selective parallels with the homologous region in humans. Together, our results represent the first reported association and functional characterization linking genetic variation and a complex trait in a natural population of nonhuman primates. PMID- 19553960 TI - Florida universities hit by funding crisis. PMID- 19553961 TI - Biology society narrows Chinese rifts. PMID- 19553968 TI - Cyberinfrastructure: Feed me data. PMID- 19553969 TI - Science journalism: Breaking the convention? PMID- 19553970 TI - Free will: it's a normal biological property, not a gift or a mystery. PMID- 19553971 TI - Free will: emotions and consciousness could contribute. PMID- 19553972 TI - Interrogation: our professional body forbids involvement. PMID- 19553973 TI - Interrogation: hard for psychologists to act as whistleblowers. PMID- 19553974 TI - How air capture could help to promote a Copenhagen solution. PMID- 19553975 TI - Planck's power lies in its unique instrument combination. PMID- 19553976 TI - Science journalism: Toppling the priesthood. PMID- 19553977 TI - Science journalism: Too close for comfort. PMID- 19553978 TI - Science journalism: The Arab boom. PMID- 19553983 TI - Structural biology: Protein dynamics from disorder. PMID- 19553984 TI - Granular media: Structures in sand streams. PMID- 19553985 TI - Cell biology: A score for membrane fusion. PMID- 19553987 TI - Planetary science: Enceladus with a grain of salt. PMID- 19553988 TI - Stem cells: The stress of forming blood cells. PMID- 19553989 TI - Nuclear physics: Unexpected doubly magic nucleus. PMID- 19553990 TI - Recent advances and emerging trends in plant hormone signalling. AB - Plant growth and development is regulated by a structurally unrelated collection of small molecules called plant hormones. During the last 15 years the number of known plant hormones has grown from five to at least ten. Furthermore, many of the proteins involved in plant hormone signalling pathways have been identified, including receptors for many of the major hormones. Strikingly, the ubiquitin proteasome pathway plays a central part in most hormone-signalling pathways. In addition, recent studies confirm that hormone signalling is integrated at several levels during plant growth and development. PMID- 19553992 TI - Sodium salts in E-ring ice grains from an ocean below the surface of Enceladus. AB - Saturn's moon Enceladus emits plumes of water vapour and ice particles from fractures near its south pole, suggesting the possibility of a subsurface ocean. These plume particles are the dominant source of Saturn's E ring. A previous in situ analysis of these particles concluded that the minor organic or siliceous components, identified in many ice grains, could be evidence for interaction between Enceladus' rocky core and liquid water. It was not clear, however, whether the liquid is still present today or whether it has frozen. Here we report the identification of a population of E-ring grains that are rich in sodium salts ( approximately 0.5-2% by mass), which can arise only if the plumes originate from liquid water. The abundance of various salt components in these particles, as well as the inferred basic pH, exhibit a compelling similarity to the predicted composition of a subsurface Enceladus ocean in contact with its rock core. The plume vapour is expected to be free of atomic sodium. Thus, the absence of sodium from optical spectra is in good agreement with our results. In the E ring the upper limit for spectroscopy is insufficiently sensitive to detect the concentrations we found. PMID- 19553991 TI - GOLPH3 modulates mTOR signalling and rapamycin sensitivity in cancer. AB - Genome-wide copy number analyses of human cancers identified a frequent 5p13 amplification in several solid tumour types, including lung (56%), ovarian (38%), breast (32%), prostate (37%) and melanoma (32%). Here, using integrative analysis of a genomic profile of the region, we identify a Golgi protein, GOLPH3, as a candidate targeted for amplification. Gain- and loss-of-function studies in vitro and in vivo validated GOLPH3 as a potent oncogene. Physically, GOLPH3 localizes to the trans-Golgi network and interacts with components of the retromer complex, which in yeast has been linked to target of rapamycin (TOR) signalling. Mechanistically, GOLPH3 regulates cell size, enhances growth-factor-induced mTOR (also known as FRAP1) signalling in human cancer cells, and alters the response to an mTOR inhibitor in vivo. Thus, genomic and genetic, biological, functional and biochemical data in yeast and humans establishes GOLPH3 as a new oncogene that is commonly targeted for amplification in human cancer, and is capable of modulating the response to rapamycin, a cancer drug in clinical use. PMID- 19553993 TI - No sodium in the vapour plumes of Enceladus. AB - The discovery of water vapour and ice particles erupting from Saturn's moon Enceladus fuelled speculation that an internal ocean was the source. Alternatively, the source might be ice warmed, melted or crushed by tectonic motions. Sodium chloride (that is, salt) is expected to be present in a long lived ocean in contact with a rocky core. Here we report a ground-based spectroscopic search for atomic sodium near Enceladus that places an upper limit on the mixing ratio in the vapour plumes orders of magnitude below the expected ocean salinity. The low sodium content of escaping vapour, together with the small fraction of salt-bearing particles, argues against a situation in which a near-surface geyser is fuelled by a salty ocean through cracks in the crust. The lack of observable sodium in the vapour is consistent with a wide variety of alternative eruption sources, including a deep ocean, a freshwater reservoir, or ice. The existing data may be insufficient to distinguish between these hypotheses. PMID- 19553994 TI - Optically controlled locking of the nuclear field via coherent dark-state spectroscopy. AB - A single electron or hole spin trapped inside a semiconductor quantum dot forms the foundation for many proposed quantum logic devices. In group III-V materials, the resonance and coherence between two ground states of the single spin are inevitably affected by the lattice nuclear spins through the hyperfine interaction, while the dynamics of the single spin also influence the nuclear environment. Recent efforts have been made to protect the coherence of spins in quantum dots by suppressing the nuclear spin fluctuations. However, coherent control of a single spin in a single dot with simultaneous suppression of the nuclear fluctuations has yet to be achieved. Here we report the suppression of nuclear field fluctuations in a singly charged quantum dot to well below the thermal value, as shown by an enhancement of the single electron spin dephasing time T(2)*, which we measure using coherent dark-state spectroscopy. The suppression of nuclear fluctuations is found to result from a hole-spin assisted dynamic nuclear spin polarization feedback process, where the stable value of the nuclear field is determined only by the laser frequencies at fixed laser powers. This nuclear field locking is further demonstrated in a three-laser measurement, indicating a possible enhancement of the electron spin T(2)* by a factor of several hundred. This is a simple and powerful method of enhancing the electron spin coherence time without use of 'spin echo'-type techniques. We expect that our results will enable the reproducible preparation of the nuclear spin environment for repetitive control and measurement of a single spin with minimal statistical broadening. PMID- 19553995 TI - High-speed tracking of rupture and clustering in freely falling granular streams. AB - Thin streams of liquid commonly break up into characteristic droplet patterns owing to the surface-tension-driven Plateau-Rayleigh instability. Very similar patterns are observed when initially uniform streams of dry granular material break up into clusters of grains, even though flows of macroscopic particles are considered to lack surface tension. Recent studies on freely falling granular streams tracked fluctuations in the stream profile, but the clustering mechanism remained unresolved because the full evolution of the instability could not be observed. Here we demonstrate that the cluster formation is driven by minute, nanoNewton cohesive forces that arise from a combination of van der Waals interactions and capillary bridges between nanometre-scale surface asperities. Our experiments involve high-speed video imaging of the granular stream in the co moving frame, control over the properties of the grain surfaces and the use of atomic force microscopy to measure grain-grain interactions. The cohesive forces that we measure correspond to an equivalent surface tension five orders of magnitude below that of ordinary liquids. We find that the shapes of these weakly cohesive, non-thermal clusters of macroscopic particles closely resemble droplets resulting from thermally induced rupture of liquid nanojets. PMID- 19553996 TI - Enhanced carbon pump inferred from relaxation of nutrient limitation in the glacial ocean. AB - The modern Eastern Equatorial Pacific (EEP) Ocean is a large oceanic source of carbon to the atmosphere. Primary productivity over large areas of the EEP is limited by silicic acid and iron availability, and because of this constraint the organic carbon export to the deep ocean is unable to compensate for the outgassing of carbon dioxide that occurs through upwelling of deep waters. It has been suggested that the delivery of dust-borne iron to the glacial ocean could have increased primary productivity and enhanced deep-sea carbon export in this region, lowering atmospheric carbon dioxide concentrations during glacial periods. Such a role for the EEP is supported by higher organic carbon burial rates documented in underlying glacial sediments, but lower opal accumulation rates cast doubts on the importance of the EEP as an oceanic region for significant glacial carbon dioxide drawdown. Here we present a new silicon isotope record that suggests the paradoxical decline in opal accumulation rate in the glacial EEP results from a decrease in the silicon to carbon uptake ratio of diatoms under conditions of increased iron availability from enhanced dust input. Consequently, our study supports the idea of an invigorated biological pump in this region during the last glacial period that could have contributed to glacial carbon dioxide drawdown. Additionally, using evidence from silicon and nitrogen isotope changes, we infer that, in contrast to the modern situation, the biological productivity in this region is not constrained by the availability of iron, silicon and nitrogen during the glacial period. We hypothesize that an invigorated biological carbon dioxide pump constrained perhaps only by phosphorus limitation was a more common occurrence in low-latitude areas of the glacial ocean. PMID- 19553997 TI - (142)Nd evidence for an enriched Hadean reservoir in cratonic roots. AB - The isotope (146)Sm undergoes alpha-decay to (142)Nd, with a half-life of 103 million years. Measurable variations in the (142)Nd/(144)Nd values of rocks resulting from Sm-Nd fractionation could therefore only have been produced within about 400 million years of the Solar System's formation (that is, when (146)Sm was extant). The (142)Nd/(144)Nd compositions of terrestrial rocks are accordingly a sensitive monitor of the main silicate differentiation events that took place in the early Earth. High (142)Nd/(144)Nd values measured in some Archaean rocks from Greenland hint at the existence of an early incompatible element-depleted mantle. Here we present measurements of low (142)Nd/(144)Nd values in 1.48-gigayear-(Gyr)-old lithospheric mantle-derived alkaline rocks from the Khariar nepheline syenite complex in southeastern India. These data suggest that a reservoir that was relatively enriched in incompatible elements formed at least 4.2 Gyr ago and traces of its isotopic signature persisted within the lithospheric root of the Bastar craton until at least 1.48 Gyr ago. These low (142)Nd/(144)Nd compositions may represent a diluted signature of a Hadean (4 to 4.57 Gyr ago) enriched reservoir that is characterized by even lower values. That no evidence of the early depleted mantle has been observed in rocks younger than 3.6 Gyr (refs 3, 4, 7) implies that such domains had effectively mixed back into the convecting mantle by then. In contrast, some early enriched components apparently escaped this fate. Thus, the mantle sampled by magmatism since 3.6 Gyr ago may be biased towards a depleted composition that would be balanced by relatively more enriched reservoirs that are 'hidden' in Hadean crust, the D'' layer of the lowermost mantle or, as we propose here, also within the roots of old cratons. PMID- 19554002 TI - Heart failure: Support for exercise training in CHF. AB - The results of HF-ACTION--the largest ever intervention trial of a nonpharmacological treatment for chronic heart failure--have been reported. The investigators randomly allocated participants to either a structured exercise program or to usual care. Although the primary end point was not reached at the prespecified significance level, the background evidence and data from previous trials and other prespecified analyses compel me to conclude that exercise training should be recommended for patients with stable chronic heart failure. PMID- 19553998 TI - Yurt, Coracle, Neurexin IV and the Na(+),K(+)-ATPase form a novel group of epithelial polarity proteins. AB - The integrity of polarized epithelia is critical for development and human health. Many questions remain concerning the full complement and the function of the proteins that regulate cell polarity. Here we report that the Drosophila FERM proteins Yurt (Yrt) and Coracle (Cora) and the membrane proteins Neurexin IV (Nrx IV) and Na(+),K(+)-ATPase are a new group of functionally cooperating epithelial polarity proteins. This 'Yrt/Cora group' promotes basolateral membrane stability and shows negative regulatory interactions with the apical determinant Crumbs (Crb). Genetic analyses indicate that Nrx-IV and Na(+),K(+)-ATPase act together with Cora in one pathway, whereas Yrt acts in a second redundant pathway. Moreover, we show that the Yrt/Cora group is essential for epithelial polarity during organogenesis but not when epithelial polarity is first established or during terminal differentiation. This property of Yrt/Cora group proteins explains the recovery of polarity in embryos lacking the function of the Lethal giant larvae (Lgl) group of basolateral polarity proteins. We also find that the mammalian Yrt orthologue EPB41L5 (also known as YMO1 and Limulus) is required for lateral membrane formation, indicating a conserved function of Yrt proteins in epithelial polarity. PMID- 19554003 TI - Stroke: Atrial fibrillation, stroke prevention therapy and aging. AB - Stroke risk in patients with nonvalvular atrial fibrillation increases markedly with age. although anticoagulation is more effective than antiplatelet therapy in atrial fibrillation, it tends to be underutilized in the elderly. A study by van Walraven et al. examines the influence of age on stroke prevention therapy in atrial fibrillation. PMID- 19554004 TI - Cardiomyopathies: New test for arrhythmogenic right ventricular cardiomyopathy. AB - Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) is challenging to diagnose because of nonspecific findings, particularly in the early phases of the disease. Clinical diagnosis is made on the basis of several criteria, but these lack sensitivity. Asimaki et al. suggest that immunohistochemical analysis of myocardial desmosomal proteins is a highly sensitive and specific diagnostic test for ARVD/C. PMID- 19554005 TI - Diabetes: Does screening for inducible ischemia improve outcomes? AB - The DIAD trial investigators have demonstrated that screening for inducible myocardial ischemia did not improve overall clinical outcome in a contemporary cohort of patients with asymptomatic type 2 diabetes, most of whom were receiving excellent medical management and few of whom underwent revascularization. Future studies should be designed to evaluate the clinical value and cost effectiveness of unconditional treatment versus screening using techniques to detect atherosclerotic and ischemic burden, coupled with therapeutic interventions, risk factor management, and selective use of revascularization in high-risk patients. PMID- 19554006 TI - Atrial fibrillation: Predicting the efficacy of pulmonary vein isolation with CMR. AB - Pulmonary vein antrum isolation is an accepted alternative to antiarrhythmic therapy for restoring normal sinus rhythm in patients with atrial fibrillation. The procedure has a high initial success rate, but many patients subsequently experience recurrence of the arrhythmia. Cardiac magnetic resonance might offer a means of identifying patients who are more likely to experience successful long term results from pulmonary vein antrum isolation. PMID- 19554007 TI - Mechanical bridge to recovery in pheochromocytoma myocarditis. AB - BACKGROUND: A 27-year-old male presented with exercise-related symptoms of chest tightness, palpitations, breathlessness and severe headache, with occasional nausea, dizziness, and blurred vision. Apart from a family history of coronary artery disease there was no other medical history of note. INVESTIGATIONS: Clinical examination, treadmill exercise test (Bruce protocol), electrocardiography, MRI of the abdomen, blood tests, chest radiography, coronary angiography, two-dimensional echocardiography, transesophageal echocardiography, microscopy of the tumor, (131)iodine metaiodobenzylguanidine scan. DIAGNOSIS: Pheochromocytoma myocarditis. MANAGEMENT: Intra-aortic balloon pump, levosimendan and dobutamine infusion, alpha-blockade with phentolamine, surgical removal of the pheochromocytoma, Levitronix (Levitronix LLC, Waltham, MA) left ventricular assist device implantation. PMID- 19554008 TI - Preventing CVD in resource-poor areas: perspectives from the 'real-world'. AB - An evolving epidemic of cardiovascular disease (CVD) is having a profound effect on the health of vulnerable populations in low-to-middle income countries with limited resources. Despite some encouraging signs (particularly initiatives from the WHO), global and regional apathy towards noncommunicable forms of CVD adds to the complexity of issues to consider when establishing cost-effective prevention programs. We present our perspective on overcoming the myriad of barriers to providing cost-effective measures for CVD prevention in a resource-poor environment through the 'prism' of our experiences in establishing the Heart of Soweto Study in South Africa. PMID- 19554009 TI - Superoxide dismutase analog (Tempol: 4-hydroxy-2, 2, 6, 6-tetramethylpiperidine 1 oxyl) treatment restores erectile function in diabetes-induced impotence. AB - We hypothesized that the administration of the superoxide dismutase (SOD) mimetic Tempol (4-hydroxy-2, 2, 6, 6-tetramethylpiperidine 1-oxyl) may reverse diabetes induced erectile dysfunction. To test this hypothesis, reactive oxygen species related genes (SOD1, SOD2, GP x 1, CAT, NOS2, NOS3) were tested, erectile functional studies and immunohistochemical analysis were carried out in diabetic rats treated with or without Tempol. Thirty Sprague-Dawley (3-4 months old) rats were divided into three groups (n=10 each), 20 with diabetes (diabetic control and Tempol treatment) and 10 healthy controls. At 12 weeks after the induction of diabetes by streptozotocin and Tempol treatment, all groups underwent in vivo cavernous nerve stimulation. Rat crura were harvested and the expression of antioxidative defense enzymes were examined by semi-quantitative reverse transcriptase PCR (RT-PCR). To confirm the RT-PCR results, we carried out immunohistochemistry (IHC) for catalase (CAT) and iNOS (NOS2). Nitration of tyrosine groups in proteins was also examined by IHC. Mean intracavernous pressure in the diabetic group was significantly lower than in the healthy controls (P <0.001) and was reversed by Tempol treatment (P <0.0108). NOS2 protein expression was significantly increased in diabetic animals compared with healthy controls and Tempol restored NOS2 protein level. Nitrotyrosine was also higher in diabetic animals and although Tempol treatment decreased its formation, it remained higher than that found in healthy controls. This study suggests that Tempol treatment increased erectile function through modulating oxidative stress related genes in diabetic rats. This is the first report about the relationship between diabetes-induced erectile dysfunction and oxidative stress, and antioxidative therapy using the superoxide dismutase mimetic, Tempol, to restore erectile function. PMID- 19554010 TI - Toxoplasmosis in pregnancy: determination of IgM, IgG and avidity in filter paper embedded blood. AB - OBJECTIVE: To detect immunoglobulin M (IgM) anti-Toxoplasma gondii antibodies and determine immunoglobulin G (IgG) titer and avidity in filter paper-embedded blood (FPEB) samples of pregnant women. STUDY DESIGN: A total of 100 FPEB samples of pregnant women (30 positive and 70 negative) were analyzed for anti-T. gondii specific IgM antibodies. Eleven and nine pairs of serum and FPEB samples were used to standardize IgG titration and avidity, respectively. Then, the correlation of avidity results was determined with 23 serum/FPEB pairs from IgG positive cases. RESULT: IgM detection in FPEB was 92% sensitive and 100% specific. The titration of IgG antibodies in FPEB correlated with that of serum (r >or=0.9). Significant difference in avidity between the acute and the undetermined/chronic cases was observed in both samples. As expected, no correlation was found between IgM levels and avidity. CONCLUSION: The FPEB is useful to infer infection phase, and thus to speed clinical decisions in congenital toxoplasmosis management. PMID- 19554011 TI - Heart rate characteristics and neurodevelopmental outcome in very low birth weight infants. AB - BACKGROUND: Sepsis in very low birth weight (VLBW) infants has been associated with an increased risk of adverse developmental outcome. We have identified abnormal heart rate characteristics (HRCs) that are predictive of impending sepsis, and we have developed a summary measure of an infant's abnormal HRCs during the neonatal hospitalization that we refer to as the cumulative HRC score (cHRC). OBJECTIVE: In this study, we tested the hypothesis that increasing cHRC is associated with an increasing risk of adverse neurodevelopmental outcome in VLBW infants. METHOD: Data were collected on 65 VLBW infants whose HRCs were monitored while in the neonatal intensive care unit and who were examined at 12 to 18 months adjusted age. Using the Bayley Scale of Infant Development-II, we identified delays in early cognitive function (i.e., Mental Developmental Index <70) and psychomotor development (i.e., Psychomotor Developmental Index <70). Cerebral palsy (CP) was diagnosed using a standard neurological examination. RESULT: Increasing cHRC score was associated with an increased risk of CP (odds ratio per 1 standard deviation increase in cHRC: 2.6, 95% confidence limits: 1.42, 5.1) and delayed early cognitive development [odds ratio: 2.3 (1.3; 4.3)]. These associations remain statistically significant when adjusted for major cranial ultrasound abnormality. There was an association of increasing cHRC and delayed psychomotor development, which did not reach statistical significance [odds ratio: 1.7 (1.0, 3.0)]. CONCLUSION: Among VLBW infants, the cumulative frequency of abnormal HRCs, which can be assessed non-invasively in the neonatal intensive care unit, is associated with an increased risk of adverse neurodevelopmental outcome. PMID- 19554012 TI - Complications of umbilical artery catheterization in a model of extreme prematurity. AB - OBJECTIVE: Umbilical artery catheter (UAC) use is common in the management of critically ill neonates; however, little information exists regarding the anatomic and vascular effects of UAC placement in premature newborns. STUDY DESIGN: Baboons were delivered at 125 days of gestation (term=185 days), treated with surfactant, had UACs placed and were ventilated for either 6 or 14 days. Animals were assigned to short-term (6 days, n=6) and long-term (14 days, n=30) UAC placement. At necropsy, aortas were removed with UACs still in place. Histological examination of upper, middle and lower aorta specimens stained with hematoxylin and eosin and immunolabeled to detect smooth muscle (alpha-actin) was carried out in a blinded manner. Controls were delivered at 125, 140 and 185 days and the aortas acquired immediately after birth. None of the non-catheterized control animals (125 days, n=4; 140 days, n=5; and 185 days, n=5) had aortic vessel thrombi or vascular wall abnormalities. RESULT: All 6 animals with short term (6/6, 100%) and 18 animals with long-term (18/30, 60%) UAC placement displayed aortic thrombi and neointimal proliferation of the vascular wall. The majority (60%) of analyzed animals with UAC placement displaying neointimal hyperplasia were immunopositive for alpha-actin, indicating the presence of smooth muscle in these lesions. CONCLUSION: Our findings suggest that both short- and long-term UAC use is associated with aortic wall pathological abnormalities compared with control animals. This study emphasizes the judicious use and early removal of UACs if possible in order to potentially prevent significant hemostatic and aortic wall vascular complications. PMID- 19554013 TI - Chest compression and/or epinephrine at birth for preterm infants <32 weeks gestational age: matched cohort study of neonatal outcomes. AB - BACKGROUND: Studies of outcomes of preterm infants after the receipt of extensive cardiopulmonary resuscitation (CPR) at birth have yielded varied results. OBJECTIVE: To compare adverse outcome (death or severe morbidities) of preterm infants <32 weeks gestational age (GA) who received chest compressions with or without administration of epinephrine at birth with those who did not receive either. DESIGN/METHOD: Data were retrospectively analyzed from a database for preterm infants <32 weeks GA discharged from hospital between July 2004 and October 2007. CASES: Infants who received chest compression with or without administration of epinephrine during the initial resuscitation. Matched cohort: Infants who did not receive extensive CPR at birth (matched for GA, sex and admission date). PRIMARY OUTCOME: Death or any of three severe morbidities (grade 3 or 4 intraventricular hemorrhage or periventricular leukomalacia; retinopathy of prematurity > stage 2 or chronic lung disease). RESULT: Sixty-six cases and 156 matched infants were identified. There were no baseline differences between groups except Apgar and severity of illness scores. Median (interquartile range) duration for chest compression (n=66) was 60 (30 to 180 s) and number of epinephrine doses (n=29) was 1 (1 to 3). Logistic regression confirmed significantly higher risk of adverse outcome among cases compared with matched controls (58 vs 37%; P=0.04, adjusted odds ratio 2.23, 95% confidence interval 1.04, 4.77). CONCLUSION: Infants born prematurely who met criteria for extensive CPR at birth experienced higher risk of combined adverse outcome, including death or severe neurological injury, severe retinopathy of prematurity or bronchopulmonary dysplasia. PMID- 19554014 TI - Methemoglobin to cumulative nitric oxide ratio and response to inhaled nitric oxide in PPHN. AB - BACKGROUND: One-third of infants with persistent pulmonary hypertension of the newborn (PPHN) do not respond to inhaled nitric oxide (iNO). If iNO is not delivered to the pulmonary vasculature because of parenchymal lung disease, it cannot interact with hemoglobin to form methemoglobin (MHb). OBJECTIVE: To study the correlation between oxygenation response to iNO in infants with PPHN secondary to parenchymal lung disease and initial MHb% to cumulative NO exposure (ppm x hours) ratio (MHb/SigmaNO). STUDY DESIGN: Retrospective chart review of neonates with PPHN secondary to parenchymal lung disease treated with iNO comparing non-responders (PaO(2)/FiO(2) ratio<10 change with iNO) with responders (>or=10 change). RESULT: Non-responders (n=16) had a PaO(2)/F(iO2) of 83+/-48 (mean+/-s.d.) and decreased to 74+/-44 after iNO. PaO(2)/FiO(2) increased from 70+/-48 to 151+/-63 with iNO among responders (n=36). The MHb/SigmaNO ratio was low (0.024+/-0.012) among non-responders compared with responders (0.07+/-0.053, P<0.005). CONCLUSION: Inadequate oxygenation response to iNO is associated with lower MHb/SigmaNO, suggesting suboptimal delivery of iNO to the pulmonary vasculature. PMID- 19554015 TI - Organism-specific platelet response and factors affecting survival in thrombocytopenic very low birth weight babies with sepsis. AB - OBJECTIVE: To study organism-specific platelet response and factors affecting survival in thrombocytopenic very low birth weight (VLBW) babies with sepsis. STUDY DESIGN: Very low birth weight babies (birth weight <1500 g) admitted to a single level-three intensive care unit from January 2000 to December 2005 were prospectively evaluated for sepsis by rapid screen test, blood counts and blood culture. In thrombocytopenic babies, organism-specific platelet response and its effect on various platelet parameters were evaluated. In addition, morbidity, mortality and factors affecting survival were studied. RESULT: Sepsis was diagnosed in 230 of 620 (37%) patients. Gram-positive sepsis occurred in 20% (46/230), Gram-negative in 71% (164/230) and fungal in 8.6% (20/230) of patients. Thrombocytopenia was observed in 67% (155/230) of babies. The frequency and duration of thrombocytopenia were more with Gram-negative and fungal infections. The incidence of persistent bacteremia, multiorgan failure and death was more in thrombocytopenic neonates (P<0.01). The incidence of multiorgan failure and death was directly related to the duration of thrombocytopenia. On multiple logistic regression analysis, poor prognostic factors include a high SNAP score at admission, a severe drop in platelet count at onset of sepsis, a low platelet nadir, a prolonged duration of thrombocytopenia, a need for platelet transfusion, less number of days off ventilation and a prolonged stay in the hospital. CONCLUSION: In thrombocytopenic VLBW babies with sepsis, organism-specific platelet response is seen. In addition, persistent bacteremia, multiorgan failure and death are more in these babies, and survival decreases with the increased severity and duration of thrombocytopenia, with prolonged ventilation and increased need for platelet transfusions. PMID- 19554016 TI - Extensive cardiopulmonary resuscitation for VLBW and ELBW infants: a systematic review and meta-analyses. AB - Studies of the outcomes of preterm infants after the receipt of extensive cardiopulmonary resuscitation (CPR) at birth or in the neonatal intensive care units (NICUs) have yielded varied results. A systematic review of the outcomes of very low birth weight (VLBW) and extremely low birth weight (ELBW) infants who received extensive resuscitation at birth or in the NICU was carried out. MEDLINE, EMBASE, CINAHL databases were searched for studies of extensive CPR in the delivery room (DR-CPR) and in NICU (NICU-CPR) that have reported neonatal or long-term outcomes. A total of 20 eligible studies were identified (11 of DR-CPR, 7 of NICU-CPR and 2 had combined data). DR-CPR was associated with an increased risk of mortality (odds ratio (OR) 2.83, 95% confidence interval (CI) 1.92, 4.16) and severe neurological injury (OR 2.27, 95% CI 1.40, 3.67) compared with infants who did not receive extensive CPR. NICU-CPR was associated with an increased risk of mortality (OR 55, 95% CI 15, 195) compared with infants who did not receive CPR; however, confidence limits were wide. The long-term outcome of survivors was reported in a limited number of studies. Extensive CPR at birth or in the NICU for VLBW or ELBW infants was associated with higher risk of mortality. PMID- 19554017 TI - Targeted depletion of Polo-like kinase (Plk) 1 through lentiviral shRNA or a small-molecule inhibitor causes mitotic catastrophe and induction of apoptosis in human melanoma cells. AB - Melanoma, one of the most lethal forms of skin cancer, remains resistant to currently available treatments. Therefore, additional target-based approaches are needed for the management of this neoplasm. Polo-like kinase 1 (Plk1) has been shown to be a crucial regulator of mitotic entry, progression, and exit. Elevated Plk1 level has been associated with aggressiveness of several cancer types and with poor disease prognosis. However, the role of Plk1 in melanoma is not well established. Here, we show that Plk1 is overexpressed in both clinical tissue specimens and cultured human melanoma cells (WM115, A375, and HS294T) when compared with normal skin tissues and cultured normal melanocytes, respectively. Furthermore, Plk1 gene knockdown through Plk1-specific shRNA or its activity inhibition by a small-molecule inhibitor resulted in a significant decrease in the viability and growth of melanoma cells without affecting normal human melanocytes. In addition, Plk1 inhibition resulted in a significant (i) decrease in clonogenic survival, (ii) multiple mitotic errors, (iii) G(2)/M cell-cycle arrest, and (iv) apoptosis of melanoma cells. This study suggests that Plk1 may have a functional relevance toward melanoma development and/or progression. We suggest that the targeting of Plk1 may be a viable approach for the treatment of melanoma. PMID- 19554018 TI - Keratinocytes function as accessory cells for presentation of endogenous antigen expressed in the epidermis. AB - The precise contribution(s) of skin dendritic cells (DCs) to immune responses in the skin has not been well delineated. We developed an intradermal (i.d.) injection model in which CD8+ T (OT-I) cells that express ovalbumin (OVA) peptide specific TCRs (Valpha2/Vbeta5) are delivered directly to the dermis of transgenic (Tg) mice expressing OVA in the epidermis. After i.d. injection, these mice reliably develop skin graft-versus-host disease (GVHD) by day 7. To determine the relative contribution of Langerhans cells (LCs) to the ensuing GVHD-like reaction, we generated K14-OVA x Langerin-diphtheria-toxin-receptor (Langerin DTR) Tg mice to allow conditional ablation of LCs in the epidermis. To delineate the role of dermal DCs (dDCs) in the reaction, we also generated K14-OVA Tg chimeras using beta(2)-microglobulin-deficient (beta(2)m) congenic donor bone marrow cells. Dermal DCs in these mice cannot present OVA to autoreactive T cells (OT-I cells), whereas the LCs are antigen presentation-competent. Unexpectedly, OT-I cell injection into diphtheria toxin (DT)-treated beta(2)m --> K14-OVA x Langerin-DTR Tg mice resulted in skin GVHD. Thus, in vivo, both LC and dDC appear to be dispensable for the induction of keratinocyte-directed, CD8-mediated effector immune responses. Furthermore and surprisingly, OVA-expressing epidermal cells depleted of LCs that could not initiate allogeneic epidermal lymphocyte reactions activated naive OT-I cells in vitro. These results indicate that keratinocytes may function as accessory cells competent to prime naive skin reactive T cells.JID JOURNAL CLUB ARTICLE: For questions, answers, and open discussion about this article, please go to http://network.nature.com/group/jidclub. PMID- 19554019 TI - Merkel cell polyomavirus in cutaneous squamous cell carcinoma of immunocompetent individuals. AB - Squamous cell carcinoma (SCC) is the second most frequently diagnosed skin cancer. It has a higher incidence in immunosuppressed individuals such as organ transplant recipients and human immunodeficiency virus (HIV) carriers. Recently, a newly described polyoma virus, Merkel cell polyomavirus (MCPyV), was found in Merkel cell carcinoma (MCC), a rare aggressive skin cancer also associated with immunosuppression. We hypothesized that MCPyV would be present in SCCs. To test for the presence of MCPyV in immunocompetent SCC patients, we used PCR primer sets directed against the large T (LT) antigen and VP1 gene of MCPyV. We detected MCPyV in 15% (26/177) of SCC DNA samples and 17% (11/63) of adjacent skin DNA samples from 21 of 58 (36%) individuals studied. We did not detect MCPyV in any matched normal blood DNA (0/57), but observed the presence of MCPyV DNA in 1 of 12 normal mouthwash DNAs. All sequenced SCC samples had a common mutation truncating the LT antigen that provides indirect evidence of viral integration. The presence of MCPyV in approximately 15% of SCCs from immunocompetent individuals warrants evaluation of MCPyV as an etiologic agent in the carcinogenesis of SCC. PMID- 19554021 TI - Analysis of extracutaneous spread in mycosis fungoides. PMID- 19554020 TI - Oral contraceptives: a risk factor for squamous cell carcinoma? AB - Oral contraceptives (OCs) affect the risk of several cancers in women, but have been virtually unstudied for squamous cell carcinoma (SCC). We examined the hypothesis that OCs influence SCC risk in a case-control study among women and also examined whether polymorphisms in the DNA repair gene, Xeroderma pigmentosum group D (XPD), modified the risk. Incident cases of SCC were identified by a network of dermatologists and pathology laboratories. Population-based controls were frequency matched to cases by age and gender (n=261 SCC cases, 298 controls). Overall, OC use was associated with a 60% higher risk of SCC (odds ratio (OR), 1.6; 95% confidence interval (95% CI): 1.0-2.5). ORs for SCC were higher among those who last used OCs > or =25 years before diagnosis (OR: 2.1; 95% CI: 1.2-3.7), and among these women, SCC risk increased with duration of use (OR for < or =2 years, 1.7; 95% CI: 0.9-3.5; OR for 3-6 years, 2.6; 95% CI: 1.0 6.5; OR for > or =7 years, 2.7; 95% CI: 0.9-8.5, P(trend)=0.01). Furthermore, the XPD Lys751Gln polymorphism was a significant modifier of the OC-SCC association (P(interaction)=0.03). These findings lead us to hypothesize a potential relationship between OCs and SCC risk, and that this could involve DNA repair pathways. PMID- 19554022 TI - Association between IL13 polymorphisms and psoriatic arthritis is modified by smoking. AB - Genetic and environmental factors influence the development of psoriasis (Ps) and psoriatic arthritis (PsA). Recently, we reported that three IL13 polymorphisms, rs1800925, rs20541, and rs848, on chromosome 5q31 conferred the risk for Ps. IL13 encodes IL-13, a Th2 cytokine, and rs1800925 and rs20541 confer risk of asthma. Further, smoking may increase the risk of developing Ps. We examined the association between IL13 polymorphisms, smoking, and PsA in two Ps sample sets genotyped for rs1800925, rs20541, and rs848. We found that the minor alleles (rs1800925*T, rs20541*A, and rs848*A) were significantly associated with protection from PsA versus controls, and that no association with Ps is seen when the PsA cases are excluded. This effect was strongest with rs1800925*T (odds ratio (OR) 0.40, P(allelic) 0.000067). The prevalence of PsA in cases with the rs1800925*CT or TT genotype is about half that of those with the CC genotype (15.5 vs 32.1%, P=0.0002). However, smoking appears to abrogate this effect (CT/TT/non-smoker, prevalence of PsA 13%, OR 0.20, P=0.0001; CT/TT/smoker, prevalence 38%, OR 0.88, P=0.74, CC/non-smoker, prevalence 42% (reference), CC/smoker prevalence 47%, OR 1.21, P=0.47). This study suggests that IL13 polymorphisms associate most strongly with PsA and that smoking may modulate this effect. PMID- 19554023 TI - Treatment of metastatic melanoma with autologous Melan-A/MART-1-specific cytotoxic T lymphocyte clones. AB - Immunotherapy by adoptive T-cell transfer aims at maximizing tumor antigen specific T-cell responses. We treated 14 patients at the metastatic stage in a phase II study with Melan-A-specific T-cell clones generated from patient blood. During the period required for T-cell clone generation, the patients were treated by dacarbazine. Every patient received a T-cell clone suspension followed by subcutaneous injections of interleukin 2 and interferon alpha. Patients were monitored until disease progression occurred. We succeeded in obtaining autologous Melan-A-specific cytotoxic T lymphocyte clones, which were highly reactive against tumor cells for all the patients. Of the 14 patients treated, six (43%) experienced an objective response (CR + PR) with long-term complete remission for two patients (1 CR for 5 years and 1 CR for 28 months). Furthermore, we showed that all the clinical responses were significantly associated with in vivo expansion of the Melan-A-specific T-cell repertoire. This phenomenon appeared to be significantly associated with clinical responses. Thus, over the course of an adoptive cell transfer, monitoring this melanoma-specific T cell expansion in patient blood appears crucial for predicting the clinical efficiency of such an immunological approach. PMID- 19554024 TI - Nestin in human skin: exclusive expression in intramesenchymal skin compartments and regulation by leptin. AB - Cutaneous nestin+ cells are of substantial interest in regenerative medicine. However, the location of nestin+ cells in situ remains controversial. We therefore sought to determine their location in female human scalp skin, using stringently controlled immunohistochemical techniques, Western blot analysis, and in situ hybridization and complementing those techniques with relative and quantitative reverse transcriptase-PCR of enzymatically digested or laser-capture microdissected human hair follicle (HF) compartments. We show here that the immunoreactivity (IR) patterns obtained with anti-nestin antibodies are highly dependent on the tissue-fixation and immunohistochemical methods used. NESTIN mRNA could not be detected within HF-associated epithelial cells in situ or in RNA extracts of the microdissected HF epithelium. Instead, NESTIN transcripts were found only in intramesenchymal skin compartments. Individual cells showing both, specific nestin IR and NESTIN mRNA were detectable in the connective-tissue sheaths of human HFs, sebaceous and sweat glands. Moreover, stimulation of organ cultured human scalp skin with the adipokine leptin increased the number of nestin+ cells in these intramesenchymal skin locations, whereas no specific nestin IR could be induced by leptin within the HF epithelium, including the bulge. Therefore, nestin expression at the gene and protein levels in human scalp skin is restricted to the periappendage mesenchyme and can be stimulated by leptin. PMID- 19554025 TI - An association study of 22 candidate genes in psoriasis families reveals shared genetic factors with other autoimmune and skin disorders. AB - Psoriasis is a common inflammatory and hyperproliferative skin disease. Recent studies have reported that common genetic factors may underlie both skin and immune-mediated disorders. We hypothesized that such genes may be involved in susceptibility to psoriasis, and undertook an association analysis of 22 candidate genes in a set of French high-risk psoriasis families. One hundred fifty-three single-nucleotide polymorphisms (SNPs) were genotyped and the transmission of alleles in nuclear families was analyzed using the FBAT (family based association test). To further investigate suggestive associations, LNM (logistic-normal models) and MQLS (modified quasi-likelihood score) methods, which take the whole pedigree structure information of families into consideration, were also applied. Our study supported the involvement of six candidate genes in susceptibility to psoriasis: SCL12A8, which belongs to the solute carrier gene family; FLG and TGM5, which are involved in epidermal differentiation; CARD15 and CYLD, which modulate the transcription factor NF-kB; and IL1RN, which encodes an IL receptor antagonist. Furthermore, we found evidence for interaction between the major risk allele, HLA-Cw6, and CARD15, CYLD, and TGM5 susceptibility alleles. Taken together, our data show that shared genetic factors may contribute to the etiology of both psoriasis and other skin or immune-mediated disorders. PMID- 19554026 TI - Genetic association of Toll-like receptor 4 with cervical cytokine concentrations during pregnancy. AB - Toll-like receptors (TLRs) are critical components of innate immunity, recognizing bacterial microorganisms and initiating local inflammatory responses. In this study, we assessed the impact of genetic variation in TLR genes on cervical concentrations of pro- and anti-inflammatory cytokines, and determined whether this relationship is influenced by bacterial vaginosis (BV). A total of 4 single nucleotide polymorphisms (SNPs) in TLR2 and 12 in TLR4 were examined for associations with 10 cervical pro- and anti-inflammatory cytokine concentrations in 91 African-American (AA) and 97 European-American (EA) women in the first trimester of pregnancy. In EAs, individuals with the TT genotype at rs1554973 (TLR4) had higher cervical concentrations of interleukin-1 beta (IL-1b) compared with those with the CT or TT genotypes (P=1.5 x 10(-5)), which remains significant after correction for multiple testing. This association was more significant in women with BV (P=5 x 10(-3)) than those without BV (P=0.02). This SNP was also associated with cervical concentrations of IL-1a, IL-6, IL-8 and IP10 (interferon-gamma-inducible protein 10) (P=6 x 10(-3), 0.03, 0.05, 6 x 10( 3), respectively). Our study demonstrates that TLR4 is an important mediator of pro-inflammatory cervical immune responses, particularly in EA women and especially in those with microbial disorders such as BV. PMID- 19554027 TI - A cross-sectional survey of relationship between serum TSH level and blood pressure. AB - It is still controversial whether subclinical hypothyroidism and euthyroidism affect blood pressure. The study aimed to explore the relationship between different levels of thyroid-stimulating hormone (TSH) and blood pressure in the participants with subclinical hypothyroidism and euthyroidism. A total of 1319 participants were administered a questionnaire survey, and their blood pressure, height and body weight measurements were taken. Blood samples were taken to test serum TSH. FT3 and FT4 were further examined if TSH was abnormal. Participants were divided into euthyroid group and subclinical hypothyroidism group. Euthyroid group was further divided into three groups: group A (TSH 0.3-0.99 mIU l(-1)), group B (TSH 1.0-1.9 mIU l(-1)) and group C (TSH 1.91-4.8 mIU l(-1)). Results showed that different levels of serum TSH had no relation with systolic blood pressure (SBP) and diastolic blood pressure (DBP). The prevalence of hypertension in subclinical hypothyroidism group was significantly higher than euthyroid group in females (41.3 vs 25.6%, P<0.05). The risk of hypertension in subclinical hypothyroidism group was significantly higher than that in the euthyroid group after adjusted for age, gender, smoking status, HOMA-IR (homoeostasis model assessment of insulin resistance) and body mass index (odds ratio (OR)=1.753, 95% confidence interval (CI) 1.067-2.879, P=0.027). This association was stronger in females (OR=3.545, 95% CI 1.576-7.975, P=0.004), but there was no statistical significance in males. Within normal range of TSH, both SBP and DBP were similar among the three groups. The prevalence and risk of hypertension were also similar among the three groups. In conclusion, the prevalence of hypertension in subclinical hypothyroidism group was significantly higher than in euthyroid group in females. Change of TSH in normal range did not affect blood pressure. PMID- 19554028 TI - Effects of aerobic training intensity on resting, exercise and post-exercise blood pressure, heart rate and heart-rate variability. AB - We aimed to investigate the effects of endurance training intensity (1) on systolic blood pressure (SBP) and heart rate (HR) at rest before exercise, and during and after a maximal exercise test; and (2) on measures of HR variability at rest before exercise and during recovery from the exercise test, in at least 55-year-old healthy sedentary men and women. A randomized crossover study comprising three 10-week periods was performed. In the first and third period, participants exercised at lower or higher intensity (33% or 66% of HR reserve) in random order, with a sedentary period in between. Training programmes were identical except for intensity, and were performed under supervision thrice for 1 h per week. The results show that in the three conditions, that is, at rest before exercise, during exercise and during recovery, we found endurance training at lower and higher intensity to reduce SBP significantly (P<0.05) and to a similar extent. Further, SBP during recovery was, on average, not lower than at rest before exercise, and chronic endurance training did not affect the response of SBP after an acute bout of exercise. The effect of training on HR at rest, during exercise and recovery was more pronounced (P<0.05) with higher intensity. Finally, endurance training had no significant effect on sympathovagal balance. In conclusion, in participants at higher age, both training programmes exert similar effects on SBP at rest, during exercise and during post-exercise recovery, whereas the effects on HR are more pronounced after higher intensity training. PMID- 19554031 TI - An immobilized nanoparticle-based platform for efficient gene knockdown of targeted cells in the circulation. AB - It is well established that specific interaction between adhesion molecules of endothelial cells and receptors on leukocytes can separate and recruit leukocytes from the bloodstream to sites of inflammation and coagulation. Previously, we showed that P-selectin can be absorbed onto the surface of a blood-compatible microrenathane tube, and the P-selectin-coated surface could successfully capture P-selectin receptor-positive stem cells from physiological shear flow in vitro and from the bloodstream in vivo. In this paper, P-selectin was covalently attached to the surface of nanoscale liposomes to create targeting nanoparticles (NPs). Small interfering RNA (siRNA) was encapsulated by these nanoscale liposomes, and the liposomes were stabilized by PEGylation with DSPE-PEG2000. Experiments showed that these P-selectin-, PEGylated-, nanoscale-liposomes (PS DSPE-PEG NPs) could be absorbed onto the inner surface of microrenathane tubing. The coated surface could specifically capture targeted cells from physiological shear flow, efficiently deliver encapsulated siRNA into adherent cells and dramatically silence the targeted gene neutrophil elastase. With this device, we create a high localized concentration for siRNA delivery in the circulatory system, providing circulating target cells adequate time to interact with therapeutic materials. SiRNA is efficaciously delivered into specific target cells, thereby providing a powerful tool for highly efficient siRNA transfection and other therapeutic materials delivery in circulation. The method should prove especially useful for diseases derived from disorders of blood cells. PMID- 19554030 TI - Targeting the mammalian target of Rapamycin to inhibit VEGF and cytokines for the treatment of primary effusion lymphoma. AB - Primary effusion lymphoma (PEL) is a fatal malignancy, which typically presents as a lymphomatous effusion that later disseminates. Rapamycin (Rapa), which targets mTOR (mammalian target of Rapa), is currently evaluated as a treatment for PEL, but the recent development of PEL in Rapa-treated post-transplant recipients questions the drug's use in PEL. Here, we used a murine model of PEL effusion that mimics the human disease to investigate the anti-PEL activity of Rapa. We found that Rapa reduces ascites accumulation and extends mouse survival. Initially, Rapa reduced PEL load compared with control mice, but most mice rapidly showed PEL progression. Levels of VEGF, which promotes vascular permeability contributing to effusion formation, were significantly reduced in ascites of Rapa-treated mice compared with controls. Expression of IL-10, the principal autocrine growth factor for PEL, was initially reduced in PEL from Rapa treated mice but rapidly increased despite treatment. We found that the hypoxic environment of ascites and Rapa cooperate in stimulating IL-10 expression in PEL, which likely contributes to the emergence of drug resistance. These results identify Rapa an effective drug to reduce PEL effusions but illustrate the rapid development of drug resistance, which likely limits the efficacy of Rapa in PEL. PMID- 19554033 TI - Translational stability for sickle gene therapy. PMID- 19554032 TI - CR1/2 is an important suppressor of Adenovirus-induced innate immune responses and is required for induction of neutralizing antibodies. AB - Human complement receptors 1 and 2 are well described as important regulators of innate and adaptive immune responses, having pivotal roles in regulating complement activation (CR1) and B-cell maturation/survival. In contrast, the role of the murine homologs of CR1 and CR2 (mCR1/2) have been primarily defined as modulating activation of the adaptive immune system, with very little evidence available about the role of mCR1/2 in regulating the innate immune responses to pathogens. In this paper, we confirm that mCR1/2 plays an important role in regulating both the innate and adaptive immune responses noted after Adenovirus (Ad)-mediated gene transfer. Our results uncovered a novel role of mCR1/2 in downregulating several complement-dependent innate immune responses. We also unveiled the mechanism underlying the complement-dependent induction of neutralizing antibodies to Ad capsids as a CR1/2-dependent phenomenon that correlates with B-cell activation. These results confirm that Ad interactions with the complement system are pivotal in understanding how to maximize the safety or potency of Ad-mediated gene transfer for both gene therapy and vaccine applications. PMID- 19554029 TI - International myeloma working group (IMWG) consensus statement and guidelines regarding the current status of stem cell collection and high-dose therapy for multiple myeloma and the role of plerixafor (AMD 3100). AB - Multiple myeloma is the most common indication for high-dose chemotherapy with autologous stem cell support (ASCT) in North America today. Stem cell procurement for ASCT has most commonly been performed with stem cell mobilization using colony-stimulating factors with or without prior chemotherapy. The target CD34+ cell dose to be collected as well as the number of apheresis performed varies throughout the country, but a minimum of 2 million CD34+ cells/kg has been traditionally used for the support of one cycle of high-dose therapy. With the advent of plerixafor (AMD3100) (a novel stem cell mobilization agent), it is pertinent to review the current status of stem cell mobilization for myeloma as well as the role of autologous stem cell transplantation in this disease. On June 1, 2008, a panel of experts was convened by the International Myeloma Foundation to address issues regarding stem cell mobilization and autologous transplantation in myeloma in the context of new therapies. The panel was asked to discuss a variety of issues regarding stem cell collection and transplantation in myeloma especially with the arrival of plerixafor. Herein, is a summary of their deliberations and conclusions. PMID- 19554034 TI - Redirecting human CD4+CD25+ regulatory T cells from the peripheral blood with pre defined target specificity. AB - Recent insight into the balance of self-tolerance and auto-aggression has raised interest in using human regulatory T (Treg) cells for adoptive immunotherapy of unlimited autoimmune diseases including type-1 diabetes, rhematoid arthritis and multiple sclerosis. The therapeutic use of Treg cells, however, is so far hampered by the inefficiency of current protocols in making them accessible for genetic manipulations. We report here that TCR/CD3 stimulation that is accompanied by extensive CD28 costimulation makes human Treg cells susceptible to retroviral gene transfer ex vivo while preserving their properties in vitro and in vivo. To show the power of genetic manipulation of human Treg cells, we engineered 'designer Treg cells' by retroviral expression of a chimeric immunoreceptor with defined specificity, which activates Treg cells in a ligand dependent manner to proliferate, to secrete high amounts of interleukin-10 and to repress an ongoing cytolytic T-cell response in vivo. The procedure in genetically modifying human Treg cells ex vivo will open a panel of applications for their use in the adoptive therapy of deregulated immune responses. PMID- 19554035 TI - Human neural stem cells overexpressing glial cell line-derived neurotrophic factor in experimental cerebral hemorrhage. AB - Recent studies have reported that glial cell line-derived growth factor (GDNF) has neurotrophic effects on the central nervous system, and the neural stem cells (NSCs) engrafted in animal models of stroke survive and ameliorate the neurological deficits. In this study, a stable human NSC line overexpressing GDNF (F3.GDNF) was transplanted next to the intracerebral hemorrhage (ICH) lesion site and a possible therapeutic effect was investigated. F3.GDNF human NSC line was transplanted into the cortex overlying the striatal ICH lesion. ICH was induced in adult mice by the unilateral injection of bacterial collagenase into the striatum. The animals were evaluated for 8 weeks with rotarod and limb placement tests. Transplanted NSCs were detected by beta-gal immunostaining with double labeling of neurofilament, microtubule associated protein-2, glial fibrillary acidic protein or human nuclear matrix antigen (HuNuMA). F3.GDNF human NSCs produced a four times higher amount of GDNF over parental F3 cells in vitro, induced behavioral improvement in ICH mice after brain transplantation and two- to threefold increase in cell survival of transplanted NSCs at 2 and 8 weeks post transplantation. In F3.GDNF-grafted ICH brain, a significant increase in the antiapoptotic protein and cell survival signal molecules, and a marked reduction in proapoptotic proteins were found as compared with control group. Brain transplantation of human NSCs overexpressing GDNF in ICH animals provided functional recovery in ICH animals, and survival and differentiation of grafted human NSCs. These results indicate that the F3.GDNF human NSCs should be of a great value as a cellular source for the cellular therapy in animal models of human neurological disorders including ICH. PMID- 19554036 TI - Comparison of rhizobacterial community composition in soil suppressive or conducive to tobacco black root rot disease. AB - Work on soils suppressive to Thielaviopsis basicola-mediated tobacco black root rot has focused on antagonistic pseudomonads to date. The role of non-Pseudomonas rhizosphere populations has been neglected, and whether they differ in black root rot-suppressive versus -conducive soils is unknown. To assess this possibility, tobacco was grown in a suppressive and a conducive soil of similar physicochemical properties, and rhizobacterial community composition was compared using a 16S rRNA taxonomic microarray. The microarray contains 1033 probes and targets 19 bacterial phyla. Among them, 398 probes were designed for Proteobacteria, Firmicutes, Actinomycetes, Cyanobacteria and Bacteroidetes genera/species known to include strains relevant for plant protection or plant growth promotion. Hierarchical clustering as well as principal component analysis of microarray data discriminated clearly between black root rot-suppressive and conducive soils. In contrast, T. basicola inoculation had no impact on rhizobacterial community composition. In addition to fluorescent Pseudomonas, the taxa Azospirillum, Gluconacetobacter, Burkholderia, Comamonas and Sphingomonadaceae, which are known to comprise strains with plant-beneficial properties, were more prevalent in the suppressive soil. Mycobacterium, Bradyrhizobium, Rhodobacteraceae, Rhodospirillum and others were more prevalent in the conducive soil. For selected taxa, microarray results were largely corroborated by quantitative PCR and cloning/sequencing. In conclusion, this work identified novel bacterial taxa that could serve as indicators of disease suppressiveness in soil-quality assessments, and it extends the range of bacterial taxa hypothesized to participate in black root rot suppression. PMID- 19554037 TI - Psychrophilic methanogenic community development during long-term cultivation of anaerobic granular biofilms. AB - Granular biomass was temporally sampled from a cold (4-15 degrees C) anaerobic bioreactor, which was inoculated with mesophilic biomass and used to treat industrial wastewater in a long-term (3.4 year) study. Data from 16S rRNA gene clone libraries, quantitative PCR and terminal restriction fragment length polymorphism analyses indicated that microbial community structure was dynamic, with shifts in the archaeal and bacterial communities' structures observed following start-up and during temperature decreases from 15 to 9.5 degrees C (phase 1). Specifically, the relative abundance of architecturally important Methanosaeta-like (acetoclastic) methanogens decreased, which was concomitant with granule disintegration and the development of a putatively psychrophilic hydrogenotrophic methanogenic community. Genetic fingerprinting suggested the development of a psychroactive methanogenic community between 4 and 10 degrees C (phase 2), which was dominated by acetogenic bacteria and Methanocorpusculum-like (hydrogenotrophic) methanogens. High levels of Methanosaeta-like acetoclastic methanogens and granular biofilm integrity were maintained during phase 2. Overall, decreasing temperature resulted in distinctly altered microbial community structure during phase 1, and the development of a less dynamic psychroactive methanogenic consortium during phase 2. Moreover, psychrophilic H(2)-oxidizing methanogens emerged as important members of the psychroactive consortia after >1200 days of low-temperature cultivation. The data suggest that prolonged psychrophilic cultivation of mesophilic biomass can establish a well functioning psychroactive methanogenic consortium, thus highlighting the potential of low-temperature anaerobic digestion technology. PMID- 19554039 TI - Exogenous glucosinolate produced by Arabidopsis thaliana has an impact on microbes in the rhizosphere and plant roots. AB - A specificity of Brassicaceous plants is the production of sulphur secondary metabolites called glucosinolates that can be hydrolysed into glucose and biocidal products. Among them, isothiocyanates are toxic to a wide range of microorganisms and particularly soil-borne pathogens. The aim of this study was to investigate the role of glucosinolates and their breakdown products as a factor of selection on rhizosphere microbial community associated with living Brassicaceae. We used a DNA-stable isotope probing approach to focus on the active microbial populations involved in root exudates degradation in rhizosphere. A transgenic Arabidopsis thaliana line producing an exogenous glucosinolate and the associated wild-type plant associated were grown under an enriched (13)CO(2) atmosphere in natural soil. DNA from the rhizospheric soil was separated by density gradient centrifugation. Bacterial (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria and Acidobacteria), Archaea and fungal community structures were analysed by DGGE fingerprints of amplified 16S and 18S rRNA gene sequences. Specific populations were characterized by sequencing DGGE fragments. Roots of the transgenic plant line presented an altered profile of glucosinolates and other minor additional modifications. These modifications significantly influenced microbial community on roots and active populations in the rhizosphere. Alphaproteobacteria, particularly Rhizobiaceae, and fungal communities were mainly impacted by these Brassicaceous metabolites, in both structure and composition. Our results showed that even a minor modification in plant root could have important repercussions for soil microbial communities. PMID- 19554040 TI - The sea-surface microlayer is a gelatinous biofilm. PMID- 19554038 TI - Species-specific structural and functional diversity of bacterial communities in lichen symbioses. AB - Lichens are generally considered as mutualisms between fungi and green algae or cyanobacteria. These partnerships allow light-exposed and long-living joint structures. The unique organization of lichens provides still unexplored environments for microbial communities. To study lichen-associated bacterial communities, we analyze samples, by a polyphasic approach, from three lichen species (Cladonia arbuscula, Lecanora polytropa and Umbilicaria cylindrica) from alpine environments. Our results indicate that bacteria can form highly structured, biofilm-like assemblages on fungal surfaces and reach considerable abundances of up to 10(8) cells per gram fresh weight. Fluorescence in situ hybridization reveals the predominance of Alphaproteobacteria. Microbial fingerprints performed by PCR-single-strand conformation polymorphism analysis using universal and group-specific primers show distinct patterns for each lichen species. Characterization of cultivable strains and presence of functional genes in the total fraction suggest the involvement of associated bacteria in nutrient cycling. Ubiquitous nifH genes, which encode the nitrogenase reductase, show a high diversity and are assigned to Alphaproteobacteria and Firmicutes, for example, Paenibacillus. Cultivable strains mainly belonging to the genera Acinetobacter, Bacillus, Burkholderia, Methylobacterium and Paenibacillus show lytic (chitinolytic, glucanolytic, and proteolytic) activities, hormone production (indole-3-acetic acid) as well as phosphate mobilization and antagonistic activity toward other microorganisms. The traditional concept of lichens has to be expanded to consider multiple bacterial partners. PMID- 19554041 TI - Mining chemistry for psychiatry. PMID- 19554043 TI - A noseful of objects. PMID- 19554044 TI - Genetics meets epigenetics: HDACs and Wnt signaling in myelin development and regeneration. PMID- 19554045 TI - Stop and go GABA. PMID- 19554046 TI - Reactive oxygen species are NOXious for neurons. PMID- 19554047 TI - Prefrontal cortex and cognitive control: motivating functional hierarchies. PMID- 19554049 TI - [Strategies for renal health: a project of the Spanish Society of Nephrology]. PMID- 19554048 TI - Adult satellite cells and embryonic muscle progenitors have distinct genetic requirements. AB - Myogenic potential, survival and expansion of mammalian muscle progenitors depend on the myogenic determinants Pax3 and Pax7 embryonically, and Pax7 alone perinatally. Several in vitro studies support the critical role of Pax7 in these functions of adult muscle stem cells (satellite cells), but a formal demonstration has been lacking in vivo. Here we show, through the application of inducible Cre/loxP lineage tracing and conditional gene inactivation to the tibialis anterior muscle regeneration paradigm, that, unexpectedly, when Pax7 is inactivated in adult mice, mutant satellite cells are not compromised in muscle regeneration, they can proliferate and reoccupy the sublaminal satellite niche, and they are able to support further regenerative processes. Dual adult inactivation of Pax3 and Pax7 also results in normal muscle regeneration. Multiple time points of gene inactivation reveal that Pax7 is only required up to the juvenile period when progenitor cells make the transition into quiescence. Furthermore, we demonstrate a cell-intrinsic difference between neonatal progenitor and adult satellite cells in their Pax7-dependency. Our finding of an age-dependent change in the genetic requirement for muscle stem cells cautions against inferring adult stem-cell biology from embryonic studies, and has direct implications for the use of stem cells from hosts of different ages in transplantation-based therapy. PMID- 19554050 TI - [Biobanks: a new tool for clinical research]. PMID- 19554051 TI - [FGF23 in chronic kidney disease and kidney post-transplant patients]. AB - Phosphatonins are regulatory factors of phosphate metabolism and the FGF23 is the best studied of them. This has produced a change in our understanding in mineral metabolism and specifically of phosphate regulation. FGF23 is a 251-amino acid factor that differs from other FGF family members by having a 71-amino acid extension on the carboxyl-terminal end of the molecule that is specific for this factor. It is primarily produced by osteocytes in bone. It has a central role in phosphate homeostasis regulation, producing phosphaturia, and in vitamin D metabolism, inhibiting its production by suppression of renal 1 Alfa hydroxylase. It is believed to have an important place in the pathogenesis of early secondary hyperparathiroidism related to chronic renal insufficiency by inhibiting renal synthesis of 1,25(OH)2D in response to its increment in blood produced to increase renal phosphate excretion and maintain phosphate balance. In CRF its serum levels seem to be independent predictors of progression to terminal renal failure. In dialysis patients the determination of its serum levels would allow to predict the results of therapy with calcitriol in the treatment of secondary hyperparathyroidism; they also seem to be independent predictors of the risk of mortality during the first year of hemodialysis. Its serum levels have also been related to the development of vascular calcifications of hand arteries but not with aortic calcifications. The exposure to excessive levels of FGF23 in the early postransplant period seems to be strongly associated with postransplant hypophophatemia more than to PTH or other phosphatonins. PMID- 19554052 TI - [Cardio-metabolic properties of manidipine: beyond lowering arterial pressure?]. AB - From its introduction in the decade of the 70's the evolution of the calcium channel blockers has allowed to resolve the uncertainty initially generated by those first generation drugs. These, are characterized by a smaller oral availability, a fast vasodilator action and a short duration of action. Manidipine arises as a dihydropyridine calcium antagonist of third generation with real additional advantages regarding to previous generations. They show high lipophilia, a more prolonged action and as well as a prolonged average life at the level of his receptor and, in addition, some theoretical advantages among others calcium antagonists, improvements on the renal function by reducing the intraglomerular pressure and microalbuminuria. Nevertheless, the clinical evaluation of these last properties still depends on the results derived from clinical trials. Besides to go deep in its role in their antihypertensive effect, we presented a brief review on new cardiometabolic aspects of these dihydropyridines calcium antagonists focusing in manidipine. PMID- 19554053 TI - [Fibrate-induced deterioration of renal function]. AB - INTRODUCTION: Fibrates represent one of the medications used to treat patients with hyperlipemia. Deterioration in renal function is not a very known adverse effect of fibric acid derivates. In the last 26 months we have detected thirteen patients with acute renal failure associated to fibrates in our outpatients' clinic. SUBJECTS AND METHODS: The aim of our study is to analyze our experience in deterioration in renal function associated to fibrates use. This is a retrospective charts review. RESULTS: From the thirteen patients (8 males/5 females) with mean age of 65.5 +/- 12.2 years, ten received Fenofibrate (FN), one Bezafibrate (BZ) and two Gemfibrozil (GF). Six cases had previously normal renal function and the seven remaining had mild chronic renal failure (CRF). The increase of serum Creatinine (Crs) value was higher than 74%. Acute renal failure was reversible in 9 patients (group 1), but the other 4 did not recover their previous renal function (group 2). The average of Crs before fibrate treatment was 1.33 +/- 0.36 mg/dl (Creatinine clearance 63.2 +/- 26.6 ml/min) and the highest average of Crs during the treatment was 2.22 +/- 0.49 mg/d (Creatinine clearance 37.3 +/- 11.9 ml/min). The average time until acute renal failure diagnosis was 6.7 +/- 5.8 months and the recovery of renal function was delayed an average of 3.8 +/- 3.5 months after fibrates withdrawn. Group 2 patients had a higuer Crs and longer time with fibrates than group 1 patients. CPK values were normal in all cases. In two patients renal biopsy was performed and no significant lesions were detected. CONCLUSION: The fibrate treatment can induce an acute renal failure. Four patients (30.8%) did not recover their basal renal function. When fibrate treatment begins a renal function should be monitored specially in patients with CRF. PMID- 19554054 TI - [Prevalence and functional effect of arteriovenous fistula calcifications, evaluated by spiral CT in chronic haemodialysis patients]. AB - INTRODUCTION: Vascular calcification is a common finding in patients (pts) with end-stage renal disease (ESRD). OBJECTIVE: The aim of this cross-sectional study was to investigate the prevalence and functional effect of native arteriovenous fistula AVF (feeding artery and/or arterialized vein) calcifications evaluated by spiral computed tomography (CT) in ESRD pts undergoing chronic hemodialysis (HD). PATIENTS AND METHOD: Forty-five upper limb AVF (radial 44.4% or brachial 55.6%, mean duration 65.3 +/- 80.9 months) without evidence of significant stenosis were evaluated by CT in 45 ESRD pts (mean age 63.8 +/- 13.1 yr; sex M: 71.1%, F: 28.9%; mean time on HD 53.1 +/- 51.9 months; diabetic nephropathy 15.6%). All AVF explorations were performed using the same multi-slice spiral CT scanner (HiSpeed Dual machine, GE Medical Systems). The severity of AVF calcifications was quantified by CT using the following criteria: grade I absence of calcifications, grade II isolated calcifications (<10 groups of calcification), grade III moderate calcifications (10-20 groups of calcification) and grade IV diffuse calcifications (>20 groups of calcification). Laboratory parameters analyzed: calcium, phosphorus, parathyroid hormone; calcium x phosphorus product was calculated. The same week of CT scanning, we evaluated AVF function measuring the blood flow rate (QA). We determined QA (1559.3 +/- 980.6 ml/min) by the Delta-H method (ABF-mode, HemaMetrics, USA) using the Crit-Line III monitor (68.9%) or by Doppler ultrasound (31.1%) performed by the same radiologist using a 5-8 MHz linear transducer (Sequoia machine, Siemens-Acuson); mean arterial pressure MAP (94.7 +/- 16.3 mmHg) was recorded simultaneous with QA. RESULTS: Most pts not showed AVF calcification by CT scan (grade I: 27/45, 60%). Forty percent of pts (18/45) demonstrated any degree of AVF calcification (grade II 13.3%, grade III 8.9%, grade IV 17.8%). Pts with brachial AVF showed higher mean QA compared to pts with radial AVF (1899.1 +/- 1131.8 versus 1134.5 +/- 516.4 ml/min, p=0.005), but MAP (91.2 +/- 15.8 versus 99.0 +/- 16.2 mmHg) and the prevalence of AVF calcification (32% versus 50%) were not different between both groups (p=0.11 and p=0.24, respectively). Pts with evidence of any calcification on CT scanning (grade II, III or IV) had higher time on HD (84.6 +/- 63.1 versus 24.6 +/- 20.0 months), higher AVF duration (97.7 +/- 89.3 versus 34.6 +/- 61.2 months) and similar QA (1488.3 +/- 678.9 versus 1606.6 +/- 1148.9 ml/min) compared with pts without AVF calcification (p=0.014, p=0.001 and p=0.69, respectively); no differences in MAP (95.4 +/- 13.8 versus 94.2 +/- 17.9 mmHg), prevalence of brachial AVF (44% versus 63%) or mineral metabolism parameters were found when comparing both groups (for all comparisons, p=NS). The same results were obtained when comparing pts with a high (grade III-IV: 26.7%) and a low (grade I-II: 73.3%) AVF calcification score, or when comparing pts with diffuse (grade IV) and without (grade I) AVF calcification. CONCLUSIONS: 1) The prevalence of AVF calcification by CT scan was 40%. 2) The AVF calcification was related with time on HD and AVF duration. 3) The function of fully developed AVF without stenosis and suitable for routine HD was not impaired by the presence of calcifications. PMID- 19554055 TI - [Determination of brain natriuretic peptide in patients undergoing different haemodiafiltration techniques]. AB - IB-type natriuretic peptide is a cardiac neurohormone secreted by the cardiac ventricules in response to ventricular dilatation so plasma BNP level correlate with left ventricular mass and dysfunction. Dialysis patients have much greater levels of BNP due to the volume overload and because of reduced renal clearance. The aim of this study was to mesure and compare the BNP levels in three groups of patients who received different hemodiafiltration techniques: Daily online hemodiafiltration (HDFOLd), on-line hemodiafiltration (HDFOL) and low convective volume hemodiafiltration (HDF). Fifteen patients were included, five in each group. Pre and postdialysis BNP leves were measured during 8 weeks. The measure was done at the beginning of the week (long period), and at the end (short period), in order to study if there were significative differences between techniques and periods. We found significative differences between predialysis BNP levels in the short period (BNPpreC) and the long period (BNPpre-L). We also found significative differences with the posdialysis BNP in both periods; BNPpre- L vs. BNPpos-L (1069+/-1031 vs. 612 +/- 540). After comparing the three techniques the study showed significative differences between BNPpreC in HDF and HDFOL compared with HDFOld. And also after dialysis between BNPpos-C in HDFOLd compared with the other techniques. CONCLUSION: Although previous papers have shown that BNP levels have limited potential for assessment of hydration in hemodialysis patients, in this study our data demonstrate that after dialysis BNP levels decline in a significative way in the long and short period and we have found that patients on daily hemodialysis show lower BNP levels, and maybe this could be explained because daily on-line haemodiafiltration patients had lower weight rise between dialysis sessions and also better haemodynamic tolerance. PMID- 19554056 TI - [Is there a relationship between cystatin C and inflammatory status, oxidative stress and other cardiovascular risk factors in non-diabetic patients with chronic kidney disease?]. AB - Cystatin C is a marker of renal function and a major cardiovascular risk factor. In the general population, cystatin C appears to be influenced by factors other than renal function alone. However, information for serum cystatin C levels in chronic kidney disease (CKD) is lacking. METHODS: We studied 52 nondiabetic patients (38 men, mean age 49 years) with CKD stage 3 (22), 4 (25) or 5 (5) who had measurements of serum cystatin C levels, estimated glomerular filtration rate (MDRD), inflammatory (C-reactive protein, interleukin-6 and fibrinogen), and oxidative markers (anti-oxidized LDL antibodies, serum paraoxonase-1 activity and concentration), left ventricular mass index by echocardiography and other cardiovascular risk factors. RESULTS: Mean cystatin C levels were 2.35 +/- 0.9 mg/l. Cystatin C was positively correlated with creatinine serum levels, estimated glomerular filtration rate, PTH levels and negatively with anti oxidized LDL antibodies. On the other hand, cystatin C was not related to inflammatory markers, serum paraoxonase-1 activity and concentration, proteinuria, HDL or LDL cholesterol, serum triglycerides, left ventricular mass index or demographic factors such as age, body mass index and blood pressure. After adjustment for PTH levels and anti- oxidized LDL antibodies, only estimated glomerular filtration rate was independently related serum cystatin C levels (beta = -0.500, p = 0.001). CONCLUSION: In nondiabetic patients with CKD, cystatin C is closely related to the degree of renal dysfunction. In contrast, inflammatory state, oxidative stress, left ventricular mass index and other cardiovascular risk factors are not related to cystatin C levels in this population. PMID- 19554057 TI - [An evaluation of pain in haemodialysis patients using different validated measurement scales]. AB - Pain in haemodialysis is very common, although frequently underdiagnosed. Chronic pain in dialysis has been scarcely evaluated, and intradialytic pain has not been specifically analyzed. Our aim was to compare intradialytic versus chronic pain characteristics in the same group of twenty-seven hemodialyzed patients, to investigate whether there were or not differences between them. Several validated scales were used: a) Analogical Visual Scale, defines pain intensity from 0, no pain, to 10, the worst pain; b) Pain Management Index, that results from subtracting pain level from analgesic use, ranging from - 3 (inadequate) to + 3 (adequate management); c) McGill Pain Questionnaire, which defines three items: pain related qualitative index, number of words chosen, and present pain intensity; and d) Brief Pain Inventory, which analyses influence of pain in patient's life, was only aplicable to evaluate chronic pain. Tests were administered firstly during the dialysis session for evaluating intradialytic pain, and another day out of the session to evaluate chronic pain. Ischemic pain was the most common during the session (37%), whereas muscle-skeletal was more frequent out of the session (77%). Prevalence of pain was higher during the session (92.5% vs 77.7%, p < 0.05). Number of weekly sessions with pain was 1.78 +/- 1.2. Analogical visual score was slightly higher during the session with respect to chronic pain (3.28 +/- 2.22 vs 2.67 +/- 2.13, p = NS). Pain Management Index scores were significantly different (intradialytic: -0.81 +/- 0.76, chronic pain: -0.12 +/- 0.94). McGill test scores were similar in both situations. Only in chronic pain, time on dialysis correlated significantly with analogical visual scores, pain related index and number of words chosen, and parathyroid hormone levels with analogical visual scores and interference to displacement score from Brief Pain Inventory. Farmacological treatment was prescribed in 11% of patients with intradialytic pain (63.1% of responders) compared to 74% for those with chronic pain (53.1% of responders). In conclusion, pain in hemodialysis is very frequent and becomes undertreated. Pain scales used have been shown to be useful in this setting. Several differences appear between intradialytic and chronic pain. Chronic pain is less frequent and intense, better treated, mainly derived from a muscle-skeletal source, and it is related to time on dialysis and to secondary hyperparathyroidism. PMID- 19554058 TI - [Growing incidence of diabetic nephropathy in the region of Badajoz during the period 1991-2006]. AB - OBJECTIVE: Diabetic nephropathy is the most common cause for starting renal repacement therapy in Spain with a steady incidence since 1997. Data on incidence of diabetic nephropathy previously to dialysis are scanty because they are not registries similar to those used for renal replacement therapy. DESIGN AND METHODS: It have been retrospectively studied the records of our hospital Nephrology outward from January, 1991 to December, 2006. Diagnosis was commonly made using clinical criteria (proteinuria plus diabetic retinopathy). There were 21 cases which did not meet theses criteria and so renal biopsy was performed. RESULTS: During this time 478 (49.7 pmp) patients were diagnosed of diabetic nephropathy (mean age 61.2 years, 50.4% women). Incidence increased from 33.3 pmp (1991) to 76.2 pmp (2006). There were not significant changes in the age of patients along the time. Other common diagnosis in diabetic patients were nefroangiosclerosis (129) and glomerulonefritis (n = 103). Survival until renal replacement therapy was 87.5% at one year and 48% at five years of follow up. CONCLUSIONS: Incidence of diabetic nephropathy seems to have increase last years specially in the patients aged 70 or older. In spite of therapeutic improvements the prognosis is still unfavourable. Less rectricted age criteria for submitting patients may have influenced these results. PMID- 19554059 TI - [Thrombosis in vascular accesses for haemodialysis: rescue treatment using invasive vascular radiological techniques]. AB - BACKGROUND: The purpose of this paper is to communicate our experience in the salvage of thrombosed haemodialysis vascular accesses using interventional radiology techniques. METHODS: In the last four years, we have treated, by radiological means, 101 thrombosed haemodialysis vascular accesses. There were 44 autologous arteriovenous fistulas (43.56%) and 57 PTFE grafts (56.44%). There were 69 men (68.3%) and 32 women (31.7%). The mean age was 67.73 years (range 33 84). The mean vascular access age was 23.79 months (range 1-132). Manual catheter directed aspiration was used. Fragmented, triturated or pushed the thrombus against the pulmonary circulation was avoided in all cases. RESULTS: 78 accesses were salvaged (77.2%). Autologous fistulas average and PTFE grafts success rate were 84.44% and 71.42% respectively. Angioplasty in one or more lesions after thromboaspiration was performed in all accesses, except six (5.9%). Metallic endoprostheses were implanted in 14 accesses (13.9%). Mean follow-up was 9 months (range 0-44). Primary patency was 42.3% +/- 5 at 6 months and 32% +/- 4 at one year. Autologous fistulas patency was better than PTFE grafts patency (p < or =0,05). CONCLUSIONS: Our results suggest thrombosed autologous arteriovenous fistulas salvage is better than PTFE grafts. This justifies interventional radiology techniques in these situations. PMID- 19554060 TI - [Results of applying the Scientific Technical Quality Programme and Constant Quality Improvement for peritoneal dialysis]. AB - INTRODUCTION: In last time it was tried to homogenize the clinical activity and to make the decisions easier. In the field of Nephrology, the Spanish Society of Nephrology has developed different guidelines that have managed an improvement in patient s monitoring. That is the reason why the Quality Working Group in Nephrology was created, whose basic working field was haemodialysis, although its collaboration with an expert group in peritoneal dialysis (PD) has allowed the developement of a Scientific Technical Quality Programme and Constant Quality Improvement in PD. MATERIAL AND METHODS: We checked the clinical histories of all the patients in PD in the course of 2008 in the Peritoneal Dialysis Unit at our institution and we evaluated all the quality indicators that were described in the Scientific Technical Quality Programme and of Constant Quality Improvement in PD. RESULTS: During 2008 a total of 41 patients were treated in the Peritoneal Dialysis Unit at our institution, 43.9% women. Incidence was 14 (51.8%) and 21.4% were diabetics. No patients cames from transplant unit and 2 came from haemodilalysis unit (7.1%). Mean age in incident population was 60 +/- 13 years and in prevalent population was 53.9 +/- 14.4 years. Mean follow-up in PD was 25.9 +/- 19.9 months. Modified Charlson comorbility index average in incident patients was 6 and in prevalent patients was 5. 70.7% were included in transplant programme and 3 were transplanted in the year s course (10.3%). There were 19 hospital admissions (rate: 0.46 admission per patient/year in risk) with a mean stay of 7.3 days (rate: 3.4 days per patient/year in risk). During 2008 6 patients leaved PD (2 transfers to haemodialysis, 3 transplants and 1 death). 16 infective peritonitis (overall rate: 1 episode every 24 months) and 23 exit side infections were reported (rate: 1 episode every 18 months). Mean Kt/V was 2,4 +/- 0.06 (92.7% of patients achieved the stablished standards). All non-anuric patients had measured residual renal function and only 1 patient did not achieve the goal of fluid output > 1000 ml/day. No patient used 3.86-4.25% bags. Stablished standards were achieve by analitic indicators with regard to epoetin resistence index, LDL- cholesterol, phosphate, calcium-phosphate product and PTH. CONCLUSIONS: The application of the Scientific Technical Quality Programme and of Constant Quality Improvement in PD has made possible to know the current situation of our unit and to raise some matters when it is necessary to insist to get a better quality in our assistance. PMID- 19554061 TI - [Scintigraphy of the intraperitoneal cavity using technetium 99m as a diagnostic technique for diaphragmatic leaks in peritoneal dialysis: regarding two cases]. AB - Pleural effusion secondary to pleuroperitoneal communication is an unusual complication of continuous ambulatory peritoneal dialysis. Many modalities have been used to diagnosis pleuroperitoneal: pleural fluid analysis, chest X- ray, Tc 99m gammagraphy, computed tomography scan and magnetic resonance image. Some of these procedures are invasive or have a high risk of induced-contrast nephrotoxicity. We present two case reports of pleuroperitoneal leak in two patients on peritoneal dialysis diagnosed with Tc-99m gammagraphy. We conclude that Tc- 99m gammagraphy is a simple, safe, non invasive, low radiation exposure and cost effective method in the assessment and evaluation of complications related to peritoneal dialysis such as pleuroperitoneal leak. PMID- 19554062 TI - [Bariatric surgery in patients with focal segmental glomerulosclerosis secondary to obesity]. AB - 2 cases of proteinuria in obese non-diabetic young males, both corresponding to focal segmental glomerulosclerosis are presented. Effective reduction of body weight by bariatric surgery was followed by sustained remission of proteinuria allowing significant reduction or total removal of blockers of the reninangiotensin- system. PMID- 19554063 TI - [Independent clinical research in Spain]. PMID- 19554064 TI - [Response to comments on the editorial "Independent Clinical Research in Spain"]. PMID- 19554065 TI - [Comment on "a discussion on quality"]. PMID- 19554066 TI - [Acute renal failure after intake of mushrooms: the orellanus syndrome]. PMID- 19554067 TI - [The importance of early diagnosis to the prognosis of immune-complex tubulointerstitial nephritis]. PMID- 19554068 TI - [Atheroembolism in transplanted kidney]. PMID- 19554069 TI - [Sudden death in patient with cholesterol atheroembolism]. PMID- 19554070 TI - [Antiphospholipid syndrome and thrombotic microangiopathy]. PMID- 19554071 TI - [Pyelonephritis in crossed-fused renal ectopia]. PMID- 19554072 TI - [Jaw claudication: could it be Wegener's granulomatosis?]. PMID- 19554073 TI - [Massive hepatic haematoma in patient on haemodialysis]. PMID- 19554074 TI - [Lithium poisoning and proteinuria in the nephrotic range]. PMID- 19554075 TI - [Not significant stenosis of renal artery in a single kidney does not counter indicate placing an endovascular aortic prosthesis]. PMID- 19554076 TI - [Plasmapheresis as a co-treatment in acute renal failure secondary to myeloma kidney]. PMID- 19554077 TI - Kisspeptins: a multifunctional peptide system with a role in reproduction, cancer and the cardiovascular system. AB - The pairing of the kisspeptins (KP) with the KISS1 (GPR54) receptor has received growing attention since the description of the receptor as a molecular switch for puberty. The role of KP and its receptor, GPR54, in puberty is the most exciting finding made in the field of reproductive biology since the discovery of Gonadotropin Releasing Hormone (GnRH) in 1970s. A significant body of evidence across several species now suggests that KISS1 (GPR54) activation is a critical point in the commencement of puberty, although further investigation is required to characterize the interaction between KP and GnRH cascade. Given such pivotal roles of kisspeptins and GPR54 as gatekeepers of reproductive function, and the proven ability of sex steroids to physiologically regulate this system, it is plausible that environmental compounds with ability to interfere oestrogen and/or androgen signaling (agonists or antagonists) may target the hypothalamic kiss 1/GPR54 system, thereby inducing functional alterations of the hypothalamic pituitary-gonadal axis. Synthetic agonists targeting KISS1 (GPR54) may represent novel therapeutic agents for the treatment of hypogonadotrophic hypogonadism in some affected individuals. The diverse multifunctional nature of the KP is beginning to unravel. The unexpected role of these peptides in puberty has raised a number of important questions that remain to be answered. PMID- 19554078 TI - Olefin Exchange-Mediated Cyclopropanation of Nitriles with Homoallylic Alcohols. AB - We report herein olefin exchange-mediated cyclopropanation of nitriles with homoallylic alcohols. The use of homoallylic alcohols is central to the successful implementation. PMID- 19554079 TI - Ultrafast Electronic and Vibrational Dynamics of Stabilized A State Mutants of the Green Fluorescent Protein (GFP): Snipping the Proton Wire. AB - Two blue absorbing and emitting mutants (S65G/T203V/E222Q and S65T at pH 5.5) of the green fluorescent protein (GFP) have been investigated through ultrafast time resolved infra-red (TRIR) and fluorescence spectroscopy. In these mutants, in which the excited state proton transfer reaction observed in wild type GFP has been blocked, the photophysics are dominated by the neutral A state. It was found that the A* excited state lifetime is short, indicating that it is relatively less stabilised in the protein matrix than the anionic form. However, the lifetime of the A* state can be increased through modifications to the protein structure. The TRIR spectra show that a large shifts in protein vibrational modes on excitation of the A* state occurs in both these GFP mutants. This is ascribed to a change in H-bonding interactions between the protein matrix and the excited state. PMID- 19554080 TI - Long-term relationships between synaptic tenacity, synaptic remodeling, and network activity. AB - Synaptic plasticity is widely believed to constitute a key mechanism for modifying functional properties of neuronal networks. This belief implicitly implies, however, that synapses, when not driven to change their characteristics by physiologically relevant stimuli, will maintain these characteristics over time. How tenacious are synapses over behaviorally relevant time scales? To begin to address this question, we developed a system for continuously imaging the structural dynamics of individual synapses over many days, while recording network activity in the same preparations. We found that in spontaneously active networks, distributions of synaptic sizes were generally stable over days. Following individual synapses revealed, however, that the apparently static distributions were actually steady states of synapses exhibiting continual and extensive remodeling. In active networks, large synapses tended to grow smaller, whereas small synapses tended to grow larger, mainly during periods of particularly synchronous activity. Suppression of network activity only mildly affected the magnitude of synaptic remodeling, but dependence on synaptic size was lost, leading to the broadening of synaptic size distributions and increases in mean synaptic size. From the perspective of individual neurons, activity drove changes in the relative sizes of their excitatory inputs, but such changes continued, albeit at lower rates, even when network activity was blocked. Our findings show that activity strongly drives synaptic remodeling, but they also show that significant remodeling occurs spontaneously. Whereas such spontaneous remodeling provides an explanation for "synaptic homeostasis" like processes, it also raises significant questions concerning the reliability of individual synapses as sites for persistently modifying network function. PMID- 19554081 TI - c-Myc accelerates S-phase and requires WRN to avoid replication stress. AB - c-Myc interacts with components of the pre-replication complex and directly regulates DNA replication [1]. However the consequences of this novel c-Myc function on cell cycle dynamics and replication-associated damage are unknown. Here, we show that c-Myc overexpression in primary human fibroblasts markedly accelerates S-phase while c-Myc deficient fibroblasts exhibit a prolonged S phase. We also show that the Werner DNA helicase protein (WRN) plays a critical role in supporting c-Myc-driven S-phase, as depletion of WRN in c-Myc overexpressing cells increases DNA damage specifically at sites of DNA synthesis. This excess DNA damage activates a "replication stress" pathway involving ATR, CHK1, CHK2, and p53, leading to rapid senescence of WRN deficient c-Myc overexpressing cells. Indeed, depletion of p53 rescues this senescence response. We propose that WRN functions to repair abnormal replication structures caused by the acceleration of DNA replication by c-Myc. This work provides an additional mechanistic explanation for c-Myc-induced DNA damage and senescence, and reveals a vulnerability of c-Myc overexpressing cells that could potentially be exploited in cancer therapy. PMID- 19554082 TI - Are markers of inflammation more strongly associated with risk for fatal than for nonfatal vascular events? AB - BACKGROUND: Circulating inflammatory markers may more strongly relate to risk of fatal versus nonfatal cardiovascular disease (CVD) events, but robust prospective evidence is lacking. We tested whether interleukin (IL)-6, C-reactive protein (CRP), and fibrinogen more strongly associate with fatal compared to nonfatal myocardial infarction (MI) and stroke. METHODS AND FINDINGS: In the Prospective Study of Pravastatin in the Elderly at Risk (PROSPER), baseline inflammatory markers in up to 5,680 men and women aged 70-82 y were related to risk for endpoints; nonfatal CVD (i.e., nonfatal MI and nonfatal stroke [n = 672]), fatal CVD (n = 190), death from other CV causes (n = 38), and non-CVD mortality (n = 300), over 3.2-y follow-up. Elevations in baseline IL-6 levels were significantly (p = 0.0009; competing risks model analysis) more strongly associated with fatal CVD (hazard ratio [HR] for 1 log unit increase in IL-6 1.75, 95% confidence interval [CI] 1.44-2.12) than with risk of nonfatal CVD (1.17, 95% CI 1.04-1.31), in analyses adjusted for treatment allocation. The findings were consistent in a fully adjusted model. These broad trends were similar for CRP and, to a lesser extent, for fibrinogen. The results were also similar in placebo and statin recipients (i.e., no interaction). The C-statistic for fatal CVD using traditional risk factors was significantly (+0.017; p<0.0001) improved by inclusion of IL-6 but not so for nonfatal CVD events (p = 0.20). CONCLUSIONS: In PROSPER, inflammatory markers, in particular IL-6 and CRP, are more strongly associated with risk of fatal vascular events than nonfatal vascular events. These novel observations may have important implications for better understanding aetiology of CVD mortality, and have potential clinical relevance. PMID- 19554083 TI - A novel biological activity of praziquantel requiring voltage-operated Ca2+ channel beta subunits: subversion of flatworm regenerative polarity. AB - BACKGROUND: Approximately 200 million people worldwide harbour parasitic flatworm infections that cause schistosomiasis. A single drug-praziquantel (PZQ)-has served as the mainstay pharmacotherapy for schistosome infections since the 1980s. However, the relevant in vivo target(s) of praziquantel remain undefined. METHODS AND FINDINGS: Here, we provide fresh perspective on the molecular basis of praziquantel efficacy in vivo consequent to the discovery of a remarkable action of PZQ on regeneration in a species of free-living flatworm (Dugesia japonica). Specifically, PZQ caused a robust (100% penetrance) and complete duplication of the entire anterior-posterior axis during flatworm regeneration to yield two-headed organisms with duplicated, integrated central nervous and organ systems. Exploiting this phenotype as a readout for proteins impacting praziquantel efficacy, we demonstrate that PZQ-evoked bipolarity was selectively ablated by in vivo RNAi of voltage-operated calcium channel (VOCC) beta subunits, but not by knockdown of a VOCC alpha subunit. At higher doses of PZQ, knockdown of VOCC beta subunits also conferred resistance to PZQ in lethality assays. CONCLUSIONS: This study identifies a new biological activity of the antischistosomal drug praziquantel on regenerative polarity in a species of free living flatworm. Ablation of the bipolar regenerative phenotype evoked by PZQ via in vivo RNAi of VOCC beta subunits provides the first genetic evidence implicating a molecular target crucial for in vivo PZQ activity and supports the 'VOCC hypothesis' of PZQ efficacy. Further, in terms of regenerative biology and Ca(2+) signaling, these data highlight a novel role for voltage-operated Ca(2+) entry in regulating in vivo stem cell differentiation and regenerative patterning. PMID- 19554084 TI - What is the optimal therapy for patients with H5N1 influenza? AB - BACKGROUND TO THE DEBATE: In a 2007 article in PLoS Medicine[10], Holger J. Schunemann and colleagues described a new process used by the World Health Organization for rapidly developing clinical management guidelines in emergency situations. These situations include outbreaks of emerging infectious diseases. The authors discussed how they developed such a "rapid advice" guideline for the pharmacological management of avian influenza A (H5N1) virus infection. The guideline recommends giving the antiviral drug oseltamivir at a dose of 75 mg twice daily for five days. In this Debate, Nicholas White argues that such dosing is inadequate, Robert Webster and Elena Govorkova say that combination antiviral therapy should be used, and Tim Uyeki reminds us that clinical care of patients with H5N1 entails much more than antiviral treatment. These issues may also apply to therapy of patients hospitalized with severe disease due to novel swine-origin influenza A (H1N1) virus infection. PMID- 19554085 TI - An economic evaluation of venous thromboembolism prophylaxis strategies in critically ill trauma patients at risk of bleeding. AB - BACKGROUND: Critically ill trauma patients with severe injuries are at high risk for venous thromboembolism (VTE) and bleeding simultaneously. Currently, the optimal VTE prophylaxis strategy is unknown for trauma patients with a contraindication to pharmacological prophylaxis because of a risk of bleeding. METHODS AND FINDINGS: Using decision analysis, we estimated the cost effectiveness of three VTE prophylaxis strategies-pneumatic compression devices (PCDs) and expectant management alone, serial Doppler ultrasound (SDU) screening, and prophylactic insertion of a vena cava filter (VCF) -- in trauma patients admitted to an intensive care unit (ICU) with severe injuries who were believed to have a contraindication to pharmacological prophylaxis for up to two weeks because of a risk of major bleeding. Data on the probability of deep vein thrombosis (DVT) and pulmonary embolism (PE), and on the effectiveness of the prophylactic strategies, were taken from observational and randomized controlled studies. The probabilities of in-hospital death, ICU and hospital discharge rates, and resource use were taken from a population-based cohort of trauma patients with severe injuries (injury severity scores >12) admitted to the ICU of a regional trauma centre. The incidence of DVT at 12 weeks was similar for the PCD (14.9%) and SDU (15.0%) strategies, but higher for the VCF (25.7%) strategy. Conversely, the incidence of PE at 12 weeks was highest in the PCD strategy (2.9%), followed by the SDU (1.5%) and VCF (0.3%) strategies. Expected mortality and quality-adjusted life years were nearly identical for all three management strategies. Expected health care costs at 12 weeks were Can$55,831 for the PCD strategy, Can$55,334 for the SDU screening strategy, and Can$57,377 for the VCF strategy, with similar trends noted over a lifetime analysis. CONCLUSIONS: The attributable mortality due to PE in trauma patients with severe injuries is low relative to other causes of mortality. Prophylactic placement of VCF in patients at high risk of VTE who cannot receive pharmacological prophylaxis is expensive and associated with an increased risk of DVT. Compared to the other strategies, SDU screening was associated with better clinical outcomes and lower costs. PMID- 19554086 TI - A combined CXCL10, CXCL8 and H-FABP panel for the staging of human African trypanosomiasis patients. AB - BACKGROUND: Human African trypanosomiasis (HAT), also known as sleeping sickness, is a parasitic tropical disease. It progresses from the first, haemolymphatic stage to a neurological second stage due to invasion of parasites into the central nervous system (CNS). As treatment depends on the stage of disease, there is a critical need for tools that efficiently discriminate the two stages of HAT. We hypothesized that markers of brain damage discovered by proteomic strategies and inflammation-related proteins could individually or in combination indicate the CNS invasion by the parasite. METHODS: Cerebrospinal fluid (CSF) originated from parasitologically confirmed Trypanosoma brucei gambiense patients. Patients were staged on the basis of CSF white blood cell (WBC) count and presence of parasites in CSF. One hundred samples were analysed: 21 from stage 1 (no trypanosomes in CSF and 5 WBC/microL) patients. The concentration of H-FABP, GSTP-1 and S100beta in CSF was measured by ELISA. The levels of thirteen inflammation related proteins (IL-1ra, IL-1beta, IL-6, IL-9, IL-10, G-CSF, VEGF, IFN-gamma, TNF-alpha, CCL2, CCL4, CXCL8 and CXCL10) were determined by bead suspension arrays. RESULTS: CXCL10 most accurately distinguished stage 1 and stage 2 patients, with a sensitivity of 84% and specificity of 100%. Rule Induction Like (RIL) analysis defined a panel characterized by CXCL10, CXCL8 and H-FABP that improved the detection of stage 2 patients to 97% sensitivity and 100% specificity. CONCLUSION: This study highlights the value of CXCL10 as a single biomarker for staging T. b. gambiense-infected HAT patients. Further combination of CXCL10 with H-FABP and CXCL8 results in a panel that efficiently rules in stage 2 HAT patients. As these molecules could potentially be markers of other CNS infections and disorders, these results should be validated in a larger multi centric cohort including other inflammatory diseases such as cerebral malaria and active tuberculosis. PMID- 19554087 TI - Association of interleukin-6 signalling with the muscle stem cell response following muscle-lengthening contractions in humans. AB - BACKGROUND: The regulation of muscle stem cells in humans in response to muscle injury remains largely undefined. Recently, interleukin-6 (IL-6) has been implicated in muscle stem cell (satellite cell)-mediated muscle hypertrophy in animals; however, the role of IL-6 in the satellite cell (SC) response following muscle-lengthening contractions in humans has not been studied. METHODOLOGY/PRINCIPAL FINDINGS: Eight subjects (age 22+/-1 y; 79+/-8 kg) performed 300 maximal unilateral lengthening contractions (3.14 rad.s(-1)) of the knee extensors. Blood and muscle samples were collected before and at 4, 24, 72, and 120 hours post intervention. IL-6, IL-6 receptor, IL-6R(alpha), cyclin D1, suppressor of cytokine signling-3 (SOCS3) mRNA were measured using quantitative RT-PCR and serum IL-6 protein was measured using an ELISA kit. JAK2 and STAT3 phosphorylated and total protein was measured using western blotting techniques. Immunohistochemical analysis of muscle cross-sections was performed for the quantification of SCs (Pax7(+) cells) as well as the expression of phosphorylated STAT3, IL-6, IL-6R(alpha), and PCNA across all time-points. The SC response, as defined by an amplification of Pax7(+) cells, was rapid, increasing by 24 h and peaking 72 h following the intervention. Muscle IL-6 mRNA increased following the intervention, which correlated strongly (R(2) = 0.89, p<0.002) with an increase in serum IL-6 concentration. SC IL-6R(alpha) protein was expressed on the fiber, but was also localized to the SC, and IL-6(+) SC increased rapidly following muscle-lengthening contractions and returned to basal levels by 72 h post intervention, demonstrating an acute temporal expression of IL-6 with SC. Phosphorylated STAT3 was evident in SCs 4 h after lengthening contraction, and the downstream genes, cyclin D1 and SOCS3 were significantly elevated 24 hours after the intervention. CONCLUSIONS/SIGNIFICANCE: The increased expression of STAT3 responsive genes and expression of IL-6 within SCs demonstrate that IL 6/STAT3 signaling occurred in SCs, correlating with an increase in SC proliferation, evidenced by increased Pax7(+)/PCNA(+) cell number in the early stages of the time-course. Collectively, these data illustrate that IL-6 is an important signaling molecule associated with the SC response to acute muscle lengthening contractions in humans. PMID- 19554088 TI - Hematocrit and mean arterial blood pressure in pre- and postmenopause women. AB - The relationship between mean arterial blood pressure (MAP) and hematocrit (Hct) was studied in pre- and postmenopause women in the city of Durango, Mexico. Premenopause women show a negative trend between parameters that is not statistically significant. MAP and Hct are directly related in postmenopause women (p < 0.01). It is proposed that that this MAP/Hct relationship is in part due to differences in endothelial function where menopause decreases the capacity of the endothelium to respond to increased blood viscosity and shears stress, leading to the increased production of vasodilator mediators to compensate for changes in blood viscosity due to changes in Hct. Comparison with a large group of postmenopause women in the city of Stockholm showed identical trends. PMID- 19554089 TI - Effects of allopurinol and vitamin E on renal function in patients with cardiac coronary artery bypass grafts. AB - BACKGROUND: Acute renal failure is a common complication of cardiac surgery, with oxidants found to play an important role in renal injury. We therefore assessed whether the supplemental antioxidant vitamin E and the inhibitor of xanthine oxidase allopurinol could prevent renal dysfunction after coronary artery bypass graft (CABG) surgery. METHODS: Of 60 patients with glomerular filtration rate (GFR) < 60 mL/min scheduled to undergo CABG surgery, 30 were randomized to treatment with vitamin E and allopurinol for 3-5 days before surgery and 30 to no treatment. Serum creatinine levels and potassium and creatinine clearances were measured preoperatively and daily until day 5 after surgery. RESULTS: The patients consisted of 31 males and 29 females, with a mean age of 63 +/- 9 years. After surgery, there were no significant differences in mean serum creatinine (1.2 +/- 0.33 vs 1.2 +/- 0.4 mg/dL; p = 0.43) concentrations, or creatinine clearance (52 +/- 12.8 vs 52 +/- 12.8 mL/min; p = 0.9). The frequency of acute renal failure did not differ in treatment group compared with control (16% vs 13%; p = 0.5). Length of stay in the intensive care unit (ICU) was significantly longer in the control than in the treated group (3.9 +/- 1.5 vs 2.6 +/- 0.7 days; p < 0.001). CONCLUSION: Prophylactic treatment with vitamin E and allopurinol had no renoprotective effects in patients with pre-existing renal failure undergoing CABG surgery. Treatment with these agents, however, reduces the duration of ICU stay. PMID- 19554090 TI - Reducing morbidity and mortality in high risk patients with statins. AB - Residual coronary heart disease remains a significant problem even after adequate statin therapy for cardiovascular risk reduction as currently recommended by the Adult Treatment Panel III (ATP-III) of the National Cholesterol Education Program (NCEP). This is particularly true for the high risk patients as defined by ATP III that includes those patients who have a greater than 20% 10-year risk of adverse cardiac events. For such patients the current goal of a low-density lipoprotein cholesterol (LDL-cholesterol) maintenance level of < or =100 mg/dL plasma appears to be suboptimal. Accumulating data from several recent randomized studies of more aggressive LDL-cholesterol reduction to levels below 70 mg/dL in the high risk patients favor acceptance of such a new lower target for LDL cholesterol using more intensive statin therapy which would affect the treatment strategy for patients with coronary heart disease pre-percutaneous intervention, metabolic syndrome, diabetes mellitus, congestive heart failure, cerebrovascular disease and chronic kidney disease. PMID- 19554092 TI - A commentary on "A young man with acute dilated cardiomyopathy associated with methylphenidate". PMID- 19554091 TI - Treatment of pediatric pulmonary hypertension. AB - Pulmonary hypertension was once thought to be a rare condition and only managed in specialized centers. Now however, with the advent of echocardiography, it is found in many clinical scenarios, in the neonate with chronic lung disease, in the acute setting in the intensive care unit, in connective tissue disease and in cardiology pre- and postoperatively. We have a better understanding of the pathological process and have a range of medication which is starting to be able to palliate this previously fatal condition. This review describes the areas that are known in this condition and those that are less familiar. The basic physiology behind pulmonary hypertension and pulmonary vascular disease is explained. The histopathologic process and the various diagnostic tools are described and are followed by the current and future therapy at our disposal. PMID- 19554093 TI - The cognitive impact of anticholinergics: a clinical review. AB - CONTEXT: The cognitive side effects of medications with anticholinergic activity have been documented among older adults in a variety of clinical settings. However, there has been no systematic confirmation that acute or chronic prescribing of such medications lead to transient or permanent adverse cognitive outcomes. OBJECTIVE: Evaluate the existing evidence regarding the effects of anticholinergic medications on cognition in older adults. DATA SOURCES: We searched the MEDLINE, OVID, and CINAHL databases from January, 1966 to January, 2008 for eligible studies. STUDY SELECTION: Studies were included if the anticholinergic activity was systematically measured and correlated with standard measurements of cognitive performance. Studies were excluded if they reported case studies, case series, editorials, and review articles. DATA EXTRACTION: We extracted the method used to determine anticholinergic activity of medications and its association with cognitive outcomes. RESULTS: Twenty-seven studies met our inclusion criteria. Serum anticholinergic assay was the main method used to determine anticholinergic activity. All but two studies found an association between the anticholinergic activity of medications and either delirium, cognitive impairment or dementia. CONCLUSIONS: Medications with anticholinergic activity negatively affect the cognitive performance of older adults. Recognizing the anticholinergic activity of certain medications may represent a potential tool to improve cognition. PMID- 19554094 TI - Eggshell membrane: a possible new natural therapeutic for joint and connective tissue disorders. Results from two open-label human clinical studies. AB - BACKGROUND: Natural Eggshell Membrane (NEM) is a novel dietary supplement that contains naturally occurring glycosaminoglycans and proteins essential for maintaining healthy joint and connective tissues. Two single center, open-label human clinical studies were conducted to evaluate the efficacy and safety of NEM as a treatment for pain and inflexibility associated with joint and connective tissue disorders. METHODS: Eleven (single-arm trial) and 28 (double-arm trial) patients received oral NEM 500 mg once daily for four weeks. The primary outcome measure was to evaluate the change in general pain associated with the treatment joints/areas (both studies). In the single-arm trial, range of motion (ROM) and related ROM-associated pain was also evaluated. The primary treatment response endpoints were at seven and 30 days. Both clinical assessments were performed on the intent-to-treat (ITT) population within each study. RESULTS: Single-arm trial: Supplementation with NEM produced a significant treatment response at seven days for flexibility (27.8% increase; P = 0.038) and at 30 days for general pain (72.5% reduction; P = 0.007), flexibility (43.7% increase; P = 0.006), and ROM-associated pain (75.9% reduction; P = 0.021). Double-arm trial: Supplementation with NEM produced a significant treatment response for pain at seven days for both treatment arms (X: 18.4% reduction; P = 0.021. Y: 31.3% reduction; P = 0.014). There was no clinically meaningful difference between treatment arms at seven days, so the Y arm crossed over to the X formulation for the remainder of the study. The significant treatment response continued through 30 days for pain (30.2% reduction; P = 0.0001). There were no adverse events reported during either study and the treatment was reported to be well tolerated by study participants. CONCLUSIONS: Natural Eggshell Membrane (NEM) is a possible new effective and safe therapeutic option for the treatment of pain and inflexibility associated with joint and connective tissue (JCT) disorders. Supplementation with NEM, 500 mg taken once daily, significantly reduced pain, both rapidly (seven days) and continuously (30 days). It also showed clinically meaningful results from a brief responder analysis, demonstrating that significant proportions of treated patients may be helped considerably from NEM supplementation. The Clinical Trial Registration numbers for these trials are: NCT00750230 and NCT00750854. PMID- 19554095 TI - Emerging treatments for postmenopausal osteoporosis - focus on denosumab. AB - The pathway of the receptor activator of the nuclear factor kappaB ligand (RANKL), RANK and osteoprotegerin (OPG) plays a central role in coupling bone formation and resorption during normal bone turnover and in a wide spectrum of diseases characterized by disturbed bone remodeling, increased bone resorption and bone destruction (osteoporosis, Paget's disease of bone, rheumatoid arthritis [RA], metastatic bone disease). Clinical trials indicate that denosumab, a RANKL specific recombinant humanized monoclonal antibody, is effective in suppressing bone resorption, resulting in increase in bone mineral density (BMD) in post menopausal women with low BMD, and has the potential to prevent progression of erosions in RA and of skeletal-related events in metastatic bone disease. The effects on fracture reduction in postmenopausal osteoporosis are awaited from the recently finished FREEDOM study. In clinical trials with denosumab, overall adverse events were similar to placebo or comparators, indicating a favorable safety profile in these diseases, which until now have been available up to 4 years, but data on long-term safety will be needed. PMID- 19554096 TI - Combination therapy with dutasteride and tamsulosin for the treatment of symptomatic enlarged prostate. AB - Benign prostatic hyperplasia (BPH) is a frequent cause of lower urinary symptoms, with a prevalence of 50% by the sixth decade of life. Hyperplasia of stromal and epithelial prostatic elements that surround the urethra cause lower urinary tract symptoms (LUTS), urinary tract infection and acute urinary retention. Medical treatments of symptomatic BPH include; 1) the 5alpha-reductase inhibitors, 2) the alpha1-adrenergic antagonists, and 3) the combination of a 5alpha-reductase inhibitor and a alpha1-adrenergic antagonist. Selective alpha1-adrenergic antagonists relax the smooth muscle of the prostate and bladder neck without affecting the detrussor muscle of the bladder wall, thus decreasing the resistance to urine flow without compromising bladder contractility. Clinical trials have shown that alpha1-adrenergic antagonists decrease LUTS and increase urinary flow rates in men with symptomatic BPH, but do not reduce the long-term risk of urinary retention or need for surgical intervention. Inhibitors of 5alpha reductase decrease production of dihydrotestosterone within the prostate resulting in decreased prostate volumes, increased peak urinary flow rates, improvement of symptoms, and decreased risk of acute urinary retention and need for surgical intervention. Interim results of the ongoing Combination of Avodart and Tamsulosin (CombAt) study have shown combination therapy with the 5alpha reductase inhibitor dutasteride and the alpha1-adrenergic antagonist tamsulosin offer significant improvements from baseline compared with either drug alone. PMID- 19554097 TI - Six-month depot formulation of leuprorelin acetate in the treatment of prostate cancer. AB - Hormonal deprivation therapy is well established for the treatment of locally advanced and metastatic prostate cancer, as well as the adjuvant treatment of some patients with localized disease. Long-acting gonadotropin releasing hormone (GnRH) agonists have become a mainstay of androgen deprivation therapy, due to their efficacy, tolerability, and convenience of use. One-month, 3-month, and 4 month depot leuprorelin formulations are well established and widely used to this end. Recently, a 6-month depot leuprorelin has been approved for use in advanced and metastatic prostate cancer patients. With similar efficacy and side effect profiles to earlier formulations, 6-month depot leuprorelin is a convenient treatment option for these patients. This review will highlight the role of GnRH agonists in the treatment of prostate cancer with a focus on the clinical efficacy, pharmacology, and patient-focused outcomes of the newer 6-month 45 mg depot leuprorelin formulation in comparison to available shorter-acting products. PMID- 19554098 TI - Wound care in the geriatric client. AB - With our aging population, chronic diseases that compromise skin integrity such as diabetes, peripheral vascular disease (venous hypertension, arterial insufficiency) are becoming increasingly common. Skin breakdown with ulcer and chronic wound formation is a frequent consequence of these diseases. Types of ulcers include pressure ulcers, vascular ulcers (arterial and venous hypertension), and neuropathic ulcers. Treatment of these ulcers involves recognizing the four stages of healing: coagulation, inflammation, proliferation, and maturation. Chronic wounds are frequently stalled in the inflammatory stage. Moving past the inflammation stage requires considering the bacterial burden, necrotic tissue, and moisture balance of the wound being treated. Bacterial overgrowth or infection needs to be treated with topical or systemic agents. In most cases, necrotic tissue needs to be debrided and moisture balance needs to be addressed by wetting dry tissue and drying wet tissue. Special dressings have been developed to accomplish these tasks. They include films, hydrocolloids, hydrogel dressings, foams, hydrofibers, composite and alginate dressings. PMID- 19554099 TI - Receptor activated Ca(2+) release is inhibited by boric acid in prostate cancer cells. AB - BACKGROUND: The global disparity in cancer incidence remains a major public health problem. We focused on prostate cancer since microscopic disease in men is common, but the incidence of clinical disease varies more than 100 fold worldwide. Ca(2+) signaling is a central regulator of cell proliferation, but has received little attention in cancer prevention. We and others have reported a strong dose-dependent reduction in the incidence of prostate and lung cancer within populations exposed to boron (B) in drinking water and food; and in tumor and cell proliferation in animal and cell culture models. METHODS/PRINCIPAL FINDINGS: We examined the impact of B on Ca(2+) stores using cancer and non cancer human prostate cell lines, Ca(2+) indicators Rhod-2 AM and Indo-1 AM and confocal microscopy. In DU-145 cells, inhibition of Ca(2+) release was apparent following treatment with Ringers containing RyR agonists cADPR, 4CmC or caffeine and respective levels of BA (50 microM), (1, 10 microM) or (10, 20, 50,150 microM). Less aggressive LNCaP cancer cells required 20 microM BA and the non tumor cell line PWR1E required 150 microM BA to significantly inhibit caffeine stimulated Ca(2+) release. BA (10 microM) and the RyR antagonist dantroline (10 microM) were equivalent in their ability to inhibit ER Ca(2+) loss. Flow cytometry and confocal microscopy analysis showed exposure of DU-145 cells to 50 microM BA for 1 hr decreased stored [Ca(2+)] by 32%. CONCLUSION/SIGNIFICANCE: We show B causes a dose dependent decrease of Ca(2+) release from ryanodine receptor sensitive stores. This occurred at BA concentrations present in blood of geographically disparate populations. Our results suggest higher BA blood levels lower the risk of prostate cancer by reducing intracellular Ca(2+) signals and storage. PMID- 19554100 TI - Gender differences in the motivational processing of babies are determined by their facial attractiveness. AB - BACKGROUND: This study sought to determine how esthetic appearance of babies may affect their motivational processing by the adults. METHODOLOGY AND PRINCIPAL FINDINGS: Healthy men and women were administered two laboratory-based tasks: a) key pressing to change the viewing time of normal-looking babies and of those with abnormal facial features (e.g., cleft palate, strabismus, skin disorders, Down's syndrome and fetal alcohol syndrome) and b) attractiveness ratings of these images. Exposure to the babies' images produced two different response patterns: for normal babies, there was a similar effort by the two groups to extend the visual processing with lower attractiveness ratings by men; for abnormal babies, women exerted greater effort to shorten the viewing time despite attractiveness ratings comparable to the men. CONCLUSIONS: These results indicate that gender differences in the motivational processing of babies include excessive (relative to the esthetic valuation) motivation to extend the viewing time of normal babies by men vs. shortening the exposure to the abnormal babies by women. Such gender-specific incentive sensitization phenomenon may reflect an evolutionary-derived need for diversion of limited resources to the nurturance of healthy offspring. PMID- 19554101 TI - A trivalent virus-like particle vaccine elicits protective immune responses against seasonal influenza strains in mice and ferrets. AB - There is need for improved human influenza vaccines, particularly for older adults who are at greatest risk for severe disease, as well as to address the continuous antigenic drift within circulating human subtypes of influenza virus. We have engineered an influenza virus-like particle (VLP) as a new generation vaccine candidate purified from the supernatants of Sf9 insect cells following infection by recombinant baculoviruses to express three influenza virus proteins, hemagglutinin (HA), neuraminidase (NA), and matrix 1 (M1). In this study, a seasonal trivalent VLP vaccine (TVV) formulation, composed of influenza A H1N1 and H3N2 and influenza B VLPs, was evaluated in mice and ferrets for the ability to elicit antigen-specific immune responses. Animals vaccinated with the TVV formulation had hemagglutination-inhibition (HAI) antibody titers against all three homologous influenza virus strains, as well as HAI antibodies against a panel of heterologous influenza viruses. HAI titers elicited by the TVV were statistically similar to HAI titers elicited in animals vaccinated with the corresponding monovalent VLP. Mice vaccinated with the TVV had higher level of influenza specific CD8+ T cell responses than a commercial trivalent inactivated vaccine (TIV). Ferrets vaccinated with the highest dose of the VLP vaccine and then challenged with the homologous H3N2 virus had the lowest titers of replicating virus in nasal washes and showed no signs of disease. Overall, a trivalent VLP vaccine elicits a broad array of immunity and can protect against influenza virus challenge. PMID- 19554102 TI - Developmental stress affects song learning but not song complexity and vocal amplitude in zebra finches. AB - Several recent studies have tested the hypothesis that song quality in adult birds may reflect early developmental conditions, specifically nutritional stress during the nestling period. Whilst all of these earlier studies found apparent links between early nutritional stress and song quality, their results disagree as to which aspects of song learning or production were affected. In this study, we attempted to reconcile these apparently inconsistent results. Our study also provides the first assessment of song amplitude in relation to early developmental stress and as a potential cue to male quality. We used an experimental manipulation in which the seeds on which the birds were reared were mixed with husks, making them more difficult for the parents to obtain. Compared with controls, such chicks were lighter at fledging; they were thereafter placed on a normal diet and had caught up by 100 days. We show that nutritional stress during the first 30 days of life reduced the birds' accuracy of song syntax learning, resulting in poorer copies of tutor songs. Our experimental manipulations did not lead to significant changes in song amplitude, song duration or repertoire size. Thus, individual differences observed in song performance features probably reflect differences in current condition or motivation rather than past condition. PMID- 19554103 TI - Female-biased sex allocation of offspring by an Apodemus mouse in an unstable environment. AB - We investigated the effects of population fluctuation on the offspring's sex allocation by a weakly polygynous mouse, Apodemus argenteus, for 3 years. In acorn-poor seasons, heavier mothers invested more in sons, and lighter mothers invested more in daughters. In acorn-rich seasons, heavier mothers invested more in daughters, and lighter mothers invested more in sons. Maternal body condition and litter size affected the sex allocation. Furthermore, there was a maternal investment trade-off between a son's birth mass and the number of daughters. Based upon the effect of population fluctuation on the lifetime reproductive success of each sex, we proposed the new "safe bet hypothesis". This hypothesis predicts that frequent and unpredictable change in female distribution, which is often caused by abrupt fall in food condition, favors female-biased maternal investment to offspring by polygynous mammals and is applicable to many small mammals inhabiting in unstable environments. PMID- 19554104 TI - Fine-scale dissection of functional protein network organization by statistical network analysis. AB - Revealing organizational principles of biological networks is an important goal of systems biology. In this study, we sought to analyze the dynamic organizational principles within the protein interaction network by studying the characteristics of individual neighborhoods of proteins within the network based on their gene expression as well as protein-protein interaction patterns. By clustering proteins into distinct groups based on their neighborhood gene expression characteristics, we identify several significant trends in the dynamic organization of the protein interaction network. We show that proteins with distinct neighborhood gene expression characteristics are positioned in specific localities in the protein interaction network thereby playing specific roles in the dynamic network connectivity. Remarkably, our analysis reveals a neighborhood characteristic that corresponds to the most centrally located group of proteins within the network. Further, we show that the connectivity pattern displayed by this group is consistent with the notion of "rich club connectivity" in complex networks. Importantly, our findings are largely reproducible in networks constructed using independent and different datasets. PMID- 19554105 TI - Waiting lists for health care in Canada. PMID- 19554106 TI - Carboxy-methyl-cellulose hydrogel-filled breast implants - an ideal alternative? A report of five years' experience with this device. AB - The controversy over the potential health risks from exposure to silicone gel from breast implants has led to extensive research to improve the safety of silicone gel-filled implants. In addition, there has been simultaneous research directed toward the development of alternative filling materials, with the goal of a breast implant that would minimize health risks and maximize cosmetic attributes. In the present study, experience with a filler material that has textural characteristics similar to that of silicone gel, but which is biodegradable and can therefore be eliminated by the body in case of implant failure, is reported.Since February 2000, the authors have been monitoring carboxy-methyl-cellulose (CMC) hydrogel breast implants as participants in a prospective clinical trial. CMC hydrogel is a biodegradable, nontoxic, nonmutagenic and viscoelastic gel, which has been in clinical use since 1984 and has been available as the Monobloc breast implant since 1994. For the present study, 122 patients who underwent surgery between February 2000 and February 2005 were evaluated. It was determined that CMC hydrogel implants have a higher radiotranslucency than silicone gel, and the integrity of this device was easy to prove by clinical examination. In case of rupture, the implant can be replaced immediately; it is easily inserted and can be placed through a small incision because of its highly elastic shell. The complication rate was very low, and the patients were highly satisfied. In terms of safety, the implant is comparable with saline, and its consistency is comparable with that of silicone gel. PMID- 19554107 TI - Replantation surgery in Quebec: The bottlenecks to rapid care. AB - INTRODUCTION: Time delays resulting in prolonged ischemia have a significant impact on the successful reattachment of amputated body parts. No studies have addressed the issues surrounding delays from the time of the accident to the start of replantation surgery. The present paper identifies the bottlenecks that prolong the time before patients are able to gain access to a replant team. METHODS: A total of 50 patients underwent microsurgical replantation, because of traumatic amputation, at a university-based hospital from 1996 to 2003. The charts were analyzed to ascertain individual time intervals from the onset of injury until the beginning of replant surgery. RESULTS: The average length of time for patients who came directly to the replant centre was 3 h 40 min before surgery began. In contrast, for those referred from outlying hospitals, the elapsed time was 6 h 21 min. CONCLUSIONS: Two major bottlenecks were found. First, for patients who were referred from other health centres, delays were due to a lack of information as to where patients could receive appropriate replant surgery. Second, delays at the replant centre were primarily due to insufficient physical and human resources in the operating room. PMID- 19554108 TI - Bear maulings treated in Calgary, Alberta: Their management and sequelae. AB - BACKGROUND: Between 1994 and 2005, seven patients underwent surgery at the Foothills Medical Centre, Calgary, Alberta, for injuries sustained in bear maulings. The purpose of the present study was to document these cases and add to the literature on the management and the potential complications of bear attacks. METHODS: Data were collected retrospectively from charts. RESULTS: Seven patients were treated for injuries ranging from lacerations and puncture wounds to fractures and avulsed tissue. On average, patients underwent three operations and spent 22 days in hospital. Mean time from attack to arrival at the trauma centre was 19 h. Irrigation, debridement and intravenous antibiotics did not prevent wound infections in two patients. Six of seven patients developed acute stress disorder, and one of these patients went on to suffer from post-traumatic stress disorder. Complications ranged from infection to pulmonary embolism. CONCLUSIONS: Bear attacks result in a spectrum of injuries. Infections and psychiatric disorders are common sequelae. PMID- 19554109 TI - Soft tissue coverage of the knee joint following burns. AB - Full-thickness burns around the knee joint are usually associated with exposure of bone, tendon or ligaments. Following excision of the deep burns, there is no viable wound bed to which skin grafts can be applied. Although vacuum-assisted closure has been used to encourage granulation tissue, exposure of the tendons and ligaments following excision usually requires muscle flaps or fasciocutaneous flaps. PMID- 19554110 TI - Facial fractures - association with ocular injuries: A 13-year review of one practice in a tertiary care centre. AB - BACKGROUND: The incidence of ocular injury associated with facial fractures has been reported to be between 0.8% and 30%. OBJECTIVE: Because of this wide range of incidences, a system to stratify the risk of ocular injury by type of facial fracture was sought. METHODS: The present study reviewed 266 patients with facial fractures to determine the risk factors for ocular injury. The anatomy of each facial fracture was classified using an orbit-centred approach according to the number of orbital walls fractured. Patients were then grouped using this classification system. Charts documenting follow-up for each patient over a minimum of one year were examined and initial emergency room presentations of patients with subsequent visual impairment were recorded. RESULTS: The incidence of severe visual impairment and blindness was 4.5% overall. Although all facial fractures (including isolated fractures of the mandible) had a notable incidence of ocular injury, an increased number of orbital wall fractures were correlated with an increased incidence of permanent visual disability. One of 147 (0.68%) patients without an orbital wall fracture sustained permanent severe ocular injury or blindness, while 13.5% (five of 37) of three-wall orbital fracture patients and 25% (four of 16) of four-wall orbital fracture patients sustained the same injury. CONCLUSIONS: The risk of ocular injury in complex facial fractures can be stratified by the degree of orbital wall involvement. However, because all patients with persistent visual impairment had ocular findings on initial examination, it is proposed that oculovisual testing is a more sensitive indicator of lasting ocular injuries than the type of facial fracture. PMID- 19554111 TI - Secondary alveolar bone grafting: An outcome analysis. AB - OBJECTIVE: To review the outcome of secondary alveolar bone grafting in unilateral and bilateral cleft lip and palate. DESIGN: A surgeon's experience, by retrospective chart review, of 70 consecutive patients at a tertiary care centre. OUTCOME MEASURE: Periapical radiographs were taken at least six months after secondary alveolar bone grafting. The Enemark grading system was used to stratify graft-take. RESULTS: In unilateral clefts, 33% were level 1, 36% were level 2, 20% were level 3 and 11% were level 4. In bilateral clefts, 29% were level 1, 50% were level 2, 14% were level 3 and 7% were level 4. There was no statistically significant difference between the level of take and the type of cleft. Complications encountered were infection (n=3), fistula (n=3), pain (n=4) and bone graft exposure that led to failure (n=2). Two patients required reoperation for bone grafting. CONCLUSIONS: The iliac crest is a good donor site with excellent results and minimal morbidity. PMID- 19554112 TI - Significance of early diagnosis of abdominal compartment syndrome in major burns. AB - Four cases of abdominal compartment syndrome in patients suffering major burns, who were treated at Hamilton General Hospital, Hamilton, Ontario, from January 1998 to June 2003, are reported. The pathophysiological changes, and the high morbidity and mortality associated with this condition are also described. The significance of early diagnosis of this syndrome is discussed. The literature is reviewed and a protocol is suggested for measuring urinary bladder pressure when managing major burns. PMID- 19554113 TI - Use of bilateral groin flaps in the closure of defects of the perineum: A case report. AB - Reconstruction of perineal and groin defects is a challenging problem. Commonly used methods of reconstruction include skin grafts and local flaps. The groin flap is a vascularized axial flap based on the superficial circumflex iliac artery arising from the femoral artery just below the inguinal ligament. Due to the location, the donor defect can usually be closed primarily, leaving an acceptable scar. The use of bilateral groin flaps for coverage of defects of the anterior perineum following excision of condylomas is reported. PMID- 19554114 TI - Reconstruction of an abdominal wall defect following wide excision of metastatic squamous cell carcinoma at the percutaneous gastrostomy site in a head and neck cancer patient. AB - Cancer metastasis to an enterostomy site is a rare and serious complication. The incidence seems to be on the rise due to increased acceptance of the percutaneous endoscopic gastrostomy procedure. Because this may be associated with synchronous distant metastasis, full metastatic workup is required. Resections of these metastatic lesions require full-thickness abdominal wall reconstruction. PMID- 19554115 TI - Re: LR Chasmar. A review of 2975 consecutive operations by one surgeon in an accredited outpatient plastic surgicentre: A Canadian experience. PMID- 19554116 TI - Re: LR Chasmar. A review of 2975 consecutive operations by one surgeon in an accredited outpatient plastic surgicentre: A Canadian experience. PMID- 19554117 TI - The 6th annual toronto breast surgery symposium, march 30, 2006, and the 36th annual symposium on aesthetic plastic surgery, march 31-april 1, 2006. PMID- 19554118 TI - 'Optimum mobility' facelift. Part 1 - the theory. AB - Traditional rhytidectomy techniques, such as the cutaneous lift, the superficial musculoaponeurotic system lift, the deep plane lift and the subperiosteal lift, are mostly differentiated by their different planes of dissection. As well, many of these techniques consider the complete mobilization of tissues a prerequisite for obtaining a satisfactory result.However, is it true that the result of a rhytidectomy is linked to the choice of the dissection plane? Also, is it true that the adequacy of the surgical mobilization of tissues is vital to the outcome? The present paper discusses the above questions and introduces a factor that is believed to be crucial to the planning and success of a rhytidectomy: facial tissue mobility. The analysis of this mobility is presented and leads to the development of three theories: 'intrinsic mobility', 'surgically induced mobility' and 'optimum mobility points'. These theories form the foundation of a rhytidectomy technique termed 'optimum mobility' facelift. PMID- 19554119 TI - 'Optimum mobility' facelift. Part 2 - the technique. AB - In the first of this two-part article on the 'optimum mobility' facelift, facial tissue mobility was analyzed, and three theories or mechanisms emerged: 'intrinsic mobility', 'surgically induced mobility' and 'optimum mobility points'.In this second part, these three theories are applied to a rhytidectomy procedure termed 'optimum mobility' facelift. Before surgery, 'optimum mobility points' are marked on the skin. During surgery, the subcutaneous dissection is kept to a minimum by carrying it out precisely to these 'optimum mobility points'. The facial tissues, with their skin and superficial musculoaponeurotic system attachments intact, are then mobilized laterally using the 'intrinsic mobility' phenomenon, and this mobilization fixed in place using mattress sutures.The 'optimum mobility' facelift is an efficient rhytidectomy technique that has a thoughtful, precise plan, a low complication rate, a fast recovery and very satisfactory results. PMID- 19554120 TI - 'Optimum mobility' facelift. PMID- 19554122 TI - A day in the operating room. PMID- 19554121 TI - Staged management of a congenital nevus of the breast and abdomen presenting in a female infant. AB - The management of congenital melanocytic nevi is controversial. The authors describe their management of a congenital nevus covering the breast and abdomen of a one-month-old girl over a 16-year period. To prevent damage to the developing breast, a staged approach was used. The nevus over the abdomen was resected first at six years of age and the remaining portion was removed after breast development was complete at 16 years of age. The breast defect was resurfaced with a full thickness skin graft from the groin. The patient was pleased with the postoperative result. This staged approach allowed an optimized aesthetic result. PMID- 19554123 TI - Changes. PMID- 19554124 TI - Operation Rainbow Canada -Unexpected middle ear findings in a small group of Philippine teenagers and young adults with unrepaired cleft palates. PMID- 19554125 TI - Unilateral cleft lip and palate: Simultaneous early repair of the nose, anterior palate and lip. AB - Unilateral cleft lip and palate is a defect involving the lip, nose and maxilla. These structures are inter-related, and simultaneous early correction of all the aspects of the defect is necessary to obtain a satisfactory result that will be maintained with growth. The surgical technique combining various procedures is presented and compared with previously published reports. PMID- 19554126 TI - Late unilateral breast enlargement after insertion of silicone gel implants: A histopathological study. AB - Late unilateral breast enlargement after the insertion of silicone gel breast implants is a very rare phenomenon. The present study reports five women who presented with this finding over the past 20 years. Three of these patients presented with late unilateral hematomas, which developed nine, 12 and 14 years, respectively, after initial breast augmentation, in the absence of any known trauma. These patients presented for treatment one, four and 12 months, respectively, after their breast enlargements initially appeared. Two of these patients had developed chronic expanding hematomas. Extensive histopathological analyses of the capsules of all three patients provided explanations for the etiologies and progressions of the findings in these patients. In each of the three patients, the etiology of the hematoma was consistent with erosion of a capsular artery. Numerous large vessels were seen within the wall of the capsules. In each case, there was a class IV capsular contracture, which could have increased the friction of the intact implant against the capsule, and there was both old (hemosiderin deposits) and new bleeding into the pocket from the capsules. This supports the concept that numerous episodes of bleeding had occurred in each case. Histopathology also demonstrated the progression of the hematomas. After four weeks, only liquefied hematoma was present, while after four months, there were both liquefied hematoma and blood clotting. The hematoma on the surface of the capsule was becoming organized peripherally, but not centrally. After 12 months, the hematoma was becoming very well organized compared with the hematoma at four months. In one of the two remaining cases, late infection was the cause of the breast enlargement. Histopathology of this capsule showed that the involved capsule was six times as thick as the other side. It also showed edema and infiltration by scattered mononuclear cells, polymorphonuclear cells, and irregular crenated and degenerating nuclei. In the final patient, chronic inflammation appeared to be the cause of the breast enlargement. The histopathology of this capsule was unique. Its inner surface showed re-epithelialization and metaplasia of the ductal epithelium to form stratified squamous epithelium with early surface keratinization. These findings are consistent with synovial metaplasia. Several areas of the fibrous portion of the capsule showed patchy loss of cellular staining with loss of nuclei, indicative of necrosis. This produced a 'washed out' staining appearance. This tissue was paucivascular and fibrotic, and showed areas of fibrinoid necrosis, suggestive of mechanical abrasion and increased pressure. PMID- 19554127 TI - Silver-coated nylon dressings for pediatric burn victims. AB - BACKGROUND: Silver dressings are a proven method for burn treatment. Current challenges associated with burn treatment include pain management and limited hospital resources. A new silver-coated nylon dressing was used at the Montreal Children's Hospital (Montreal, Quebec) to help reduce traumatic dressing changes and cost. METHODS: Burn victims in a pediatric patient population were followed over two years. Patients were excluded if they were evaluated more than 48 h postburn or if the burn affected less than 5% of the total body surface area. The same burn team admitted and treated all case subjects, and one dressing nurse recorded and monitored all progress throughout the study to ensure standardization. RESULTS: Fifteen patients were included in the study. The average number of dressing changes needed was 4.13, with a median of three changes. The average total body surface area burned was 8%, with a mean of 13.9 days before superficial wounds were re-epithelialized. The average length of in hospital stay was four days. The cost was $388 less for silver-coated nylon dressings than for silver sulfadiazine cream for seven days of treatment. Silver coated nylon dressings did not leave any residue or pseudoeschar on the wounds and were easily maintained at home. CONCLUSION: The silver-coated nylon dressings are as effective as other silver dressings used for pediatric burn victims. The dressings are less traumatic, require fewer resources and do not leave wound residue compared with other dressings. PMID- 19554128 TI - Rate of pediatric tap water scald injuries in eastern Ontario. AB - BACKGROUND: Tap water scalds can be devastating and are entirely preventable. This topic has received notable attention in Canada in recent years, with a national campaign in 2001 directed at parents recommending that they lower the water temperature of their home water heater. This campaign has been evaluated and reported as a successful population-based intervention. OBJECTIVES: The present study evaluated the rate of pediatric tap water scalds in Ottawa, Ontario over the 10-year period from 1993 to 2002 in an attempt to identify the impact of this national awareness campaign. METHODS: Data from the Children's Hospital of Eastern Ontario available in the Canadian Hospitals Injury Reporting and Prevention Program database were used to calculate the yearly incidence density of tap water scalds among children younger than 15 years in the city of Ottawa. RESULTS: It was found that there was no statistically significant change in the rate of pediatric tap water scalds following the national campaign, and that the rate of these injuries was already very low (3.73 per 100,000 person-years over the study period). Of these, only 17% required observation or admission to hospital. CONCLUSIONS: Given the low incidence of these injuries, their priority as a public health concern needs to be re-evaluated. If they continue to be a priority, new preventive strategies need to be used. It is thought that resources used to lobby for legislation of lower hot water temperatures may be more effective in reducing the incidence of these injuries. PMID- 19554129 TI - Evaluation of sun protection behaviour in patients following excision of a skin lesion. AB - The present prospective study evaluated sun exposure and sun protection behaviours in patients diagnosed with a precancerous or cancerous skin lesion. Following signed informed consent, patients were given a questionnaire regarding sun protection and sun exposure behaviour. There were 50 patients (27 men, 23 women) with a mean age of 59 years. Following diagnosis of a cancerous or precancerous lesion, there was a significant increase (P<0.001) in sunscreen use, use of sun protection factor greater than 30, wearing a hat and avoidance of sun between 10:00 and 14:00. There were significantly more women who used sunscreen (P=0.04) and limited daily activities (P=0.03). There were significantly more patients with nonmelanoma lesions who wore hats (P<0.04) and more patients with melanoma who limited daily activities (P<0.04). While many patients in the present study did alter their sun protection and exposure behaviour, there were many patients who continued in risk-taking sun behaviour. Therefore, patients with precancerous or cancerous skin lesions who are not compliant with sun protective behaviour may benefit from more comprehensive patient education to encourage better sun protection behaviour. PMID- 19554130 TI - Vertical mammaplasty: Postoperative changes, complications and patient evaluation. AB - The aim of the present study was to investigate postoperative changes after vertical mammaplasty. Between 2002 and 2005, 72 consecutive patients aged 15 to 69 years with an average weight of 72 kg underwent bilateral vertical mammaplasty. Forty-two patients attended the regular follow-up one week, four weeks, three months, six months and one year after the operation. Nipple diameter, notch-to-nipple distance, scar length and the number of skin folds along the vertical scar were evaluated. Complications were recorded during the entire follow-up period. A questionnaire was used to document patient satisfaction 12 months after the operation. The main changes took place during the first three months after surgery. Nipple diameter showed an average increase of 28% after surgery, and the notch-to-nipple distance increased by an average of 17% over the intraoperative value. The average increase of the scar length after one year was 22%. The rate of complications was low, and patient satisfaction was high. PMID- 19554131 TI - Anomalous flexor digitorum superficialis muscle belly presenting as a mass within the palm. AB - Anomalies of the flexor digitorum superficialis muscle are extremely uncommon and usually present as a painful mass or pseudotumour within the palm. Diagnosis may be difficult because many other soft tissue tumours (lipomas, ganglions, giant cell tumours and hamartomas) may present similarly. Magnetic resonance imaging helps to define the extent and characteristics of this anomalous muscle belly and to distinguish it from a soft tissue sarcoma, whereas plain radiographs are of little value. Three types of flexor digitorum superficialis muscle anomalies have been described, and treatment consists of subtotal or total surgical debulking of the mass if symptoms persist or if the diagnosis is in question. Most patients have complete resolution and full recovery. To date, 20 cases have been reported in the literature, usually involving the right small finger. In the present paper, the case of an anomalous flexor digitorum superficialis muscle in a 17 year-old male patient's left index finger is reported. Symptoms were relieved following surgical debulking and hand-based occupational therapy. PMID- 19554133 TI - A bilobed groin flap for coverage of traumatic injury to both the volar and dorsal hand surfaces. AB - BACKGROUND: There are many methods available for coverage of both the volar and dorsal hand surfaces in traumatic injury. All of these surgical procedures allow for ample coverage of the defect, but have the major drawback of needing multiple donor sites. In the present report, a case of a complex crush injury to the hand in which both volar and dorsal surfaces received traumatic injury is presented. A bilobed groin flap pedicled on the superficial circumflex iliac artery was fashioned to cover the entire defect. Like many other described flaps, the bilobed groin flap resulted in a favourable functional and cosmetic result for the patient, but with the unique advantage of requiring a single donor site. METHODS: A bipedicled groin flap was raised from distal to proximal, and the flaps inset onto both the dorsal and volar hand defects. RESULTS: The patient regained adequate function of his hand, and was able to return to work full-time as a manual labourer. CONCLUSION: The bilobed groin flap appears to be a valuable option for covering complex hand injuries involving both the volar and dorsal surfaces of the hand. PMID- 19554132 TI - Basal cell carcinoma masquerading as a hallux valgus. AB - The incidence of primary skin cancers of the foot is exceedingly low; conversely, problems associated with a hallux valgus are common. A nonhealing ulcer overlying a hallux valgus managed conservatively with ointments and orthotic adjustments, and even with skin grafts, did not resolve over a period of 10 years. Ultimately, a shave biopsy revealed that the lesion was a basal cell carcinoma. Wide local excision and another skin graft resulted in tumour eradication and, finally, healing. Basal cell carcinoma associated with a hallux valgus has not been previously reported, and this reinforces the concept that malignant degeneration as the cause of any chronic ulceration should not be overlooked. PMID- 19554135 TI - Consent can be made easier. PMID- 19554134 TI - Orthognathic surgery for management of Arthrogryposis Multiplex Congenita: Case report and review of the literature. AB - Arthrogryposis Multiplex Congenita is a condition characterized by multiple contractures of the joints. Involvement of the temporomandibular joint is a common complication that limits mandibular opening. A case of Arthrogryposis Multiplex Congenita with anterior open bite and limited maximal incisal opening is presented. Orthognathic surgery, consisting of segmental LeFort I osteotomy and bilateral sagittal split osteotomy, was performed successfully on this patient. PMID- 19554136 TI - Application of the CONSORT statement to randomized controlled trials comparing endoscopic and open carpal tunnel release. AB - BACKGROUND: The CONSORT (Consolidated Standards of Reporting Trials) statement was developed by a group of clinical trialists, biostatisticians, epidemiologists and biomedical editors as a means to improve the quality of reports of randomized controlled trials (RCTs). The purpose of the present study is to assess the reporting quality of published RCTs that compare endoscopic carpal tunnel release (ECTR) with open carpal tunnel release (OCTR) using the CONSORT statement. METHODS: A computerized literature search was conducted to identify all RCTs published from January 1989 to November 2004 that compared ECTR with OCTR. Foreign language studies were also included, and translated versions of these studies were obtained. Two investigators independently reviewed each eligible article and determined whether the authors reported on each of the 22 items of the CONSORT statement. Disagreements were resolved by consensus. The mean scores for studies published before the introduction of the CONSORT statement and those published afterward were compared. Similarly, a comparison was made between foreign language studies and those published in English. RESULTS: Eighteen RCTs comparing ECTR with OCTR met the inclusion criteria. The total scores on the CONSORT checklist ranged from 3 to 20, with a mean score of 9.83+/-3.79 (the maximum possible score was 22). The six studies published in foreign language journals had a statistically significantly lower mean score than the studies published in English language journals (7.00+/-2.76 versus 11.25+/-3.49, respectively; P<0.05). The mean score was higher for studies published after 1996 than for those published in 1996 or earlier (12.14+/-3.80 versus 8.36+/-3.11, respectively; P<0.05). CONCLUSIONS: The quality of reporting improved over time, but no study met all 22 criteria of the CONSORT statement. The CONSORT scores were higher for studies published after 1996 and for studies published in English language journals. Despite the improvement after 1996, most of these RCTs only reported one-half of the items listed on the CONSORT statement. Future investigators of surgical RCTs should make an effort to comply with the CONSORT checklist. PMID- 19554137 TI - The epidemiology of cutaneous malignant melanoma in Nova Scotia. AB - BACKGROUND: Since 1993, the annual increase in cutaneous malignant melanoma (MM) incidence has been one of the highest for all cancers registered in Canada, with the leading rate in Nova Scotia (NS). The purpose of the present study was to document the pathological and epidemiological data on MM cases found in NS. PATIENTS AND METHODS: All MM cases identified by the Nova Scotia Cancer Registry from January 1998 to December 2002 were evaluated. The five-year survival outlook, by major prognostic factors, was also determined. In addition, the annual incidence and mortality rates from 1972 to 2002 were computed. RESULTS: Between 1998 and 2002, 925 MM cases were recorded. The age-standardized incidence rate for males and females in this period was 19.2 and 16.1 per 100,000 respectively. Men 65 years of age or older had the highest age-specific rate. The most common MM had a Breslow's depth of less than 1.0 mm (61.9%) and was Clark's level II (34.9%). There was no significant seasonal variation noted in the time of diagnosis. Survival analyses indicated that sex, age, tumour location and thickness were significant independent predictors. Despite the increase in incidence, there have only been modest changes in the annual mortality rate. CONCLUSION: The incidence of MM in NS increases with age, and is nearly double for men 65 years of age or older, compared with women in the same age group. Thin melanomas on the extremities of young females have the best prognosis in NS, which is similar to other parts of the world. Incidence appears to be unrelated to season. Public health interventions are necessary to reduce the burden of this disease. PMID- 19554138 TI - The Essex-Lopresti injury: More than just a pain in the wrist. AB - As the scope of plastic surgical practice expands to include disorders of the carpus and wrist, it has become increasingly important for plastic surgeons to understand pathoanatomy that has not traditionally been considered an integral component of training. The Essex-Lopresti injury consists of a radial head fracture with associated injury to the forearm interosseus membrane and longitudinal instability of the distal radioulnar joint. Early recognition of this disorder usually results in a predictable and satisfactory outcome. However, when this disorder is unrecognized, late reconstruction is challenging and unpredictable, and treatment may be misdirected to the wrist alone if the forearm and elbow are not considered as a component of this injury. The present report describes the importance of examining the elbow in all cases of wrist pain. As well, the literature is reviewed regarding the differences in treatment of acute and chronic Essex-Lopresti injuries. As plastic surgeons become more involved in the treatment of wrist injuries, the conscientious practitioner should be aware of more complicated pathology that may present as a seemingly straightforward wrist problem. PMID- 19554139 TI - Autoinflation of saline-filled inflatable breast implants. AB - Spontaneous autoinflation of saline-filled breast implants is a rare phenomenon; only 20 cases have been reported in the world literature. Over the past seven years, three patients have presented with significant unilateral autoinflation of their smooth, single-lumen, round, saline-filled implants. This developed at various times: progressively over 23 years with a Simaplast implant; between the ninth and 10th year after augmentation with a leaflet valve implant; and slowly over the first four years with a leaflet valve implant. The etiology of the autoinflation was shown to be different for the two types of implants. The Simaplast implant had likely been injected with a hypertonic filling solution - one that was twice as concentrated as 'normal saline'. This would have created an osmotic gradient, which would have facilitated autoexpansion by diffusion. The implant solution remained clear and transparent. In addition, there were no detectable levels of glucose, uric acid or albumin in the fluid. By contrast, auto-inflation of the leaflet valve implants likely resulted from mechanical alterations of the valve mechanism. This type of implant is known to have a high deflation rate, with frequent partial deflations. It is interesting that one of the patients with the leaflet valve implants presented with an autoexpansion on one side and a partial deflation on the other side. Both implants were from the same lot number. The same mechanism that caused partial deflations may have also allowed fluid from the implant pocket to pass through the valve into the lumen of these implants. This could allow glucose, protein and cellular elements to enter into the lumen (these would not pass through an intact elastomeric shell) which would create an osmotic gradient, allowing water to enter the elastomeric shell by diffusion. The fluid in these leaflet valve implants was brownish yellow, very viscous and turbid. It contained elevated levels of glucose and uric acid which would not have passed through the elastomeric shell. Over the past 10 years, four different theories have been proposed in an attempt to explain the etiology of autoinflation. However, the findings of the present study indicate that there are only two mechanisms - a hypertonic filling solution and alterations of the valve mechanism. PMID- 19554141 TI - Scalp expansion with the Canica Wound Closure System: First case report. AB - The closure of difficult wounds has been an ongoing challenge to the plastic and reconstructive surgeon. New techniques and technology have created innovative methods for this clinical and surgical problem. The Canica Wound Closure System is an external system of tissue expansion that has previously been shown to be successful for delayed abdominal and extremity fasciotomy closures. The hair bearing scalp is unique in that no other tissue in the human body can adequately simulate it. The first known case in North America of a patient in whom the Canica Wound Closure System was used for the successful closure of a large scalp defect following a resection for a Clark level V melanoma is presented. PMID- 19554140 TI - Diagnosis of finger flexor pulley injury in rock climbers: A systematic review. AB - BACKGROUND: Closed injury to the finger flexor pulley system is found frequently in rock climbers. There are no evidence-based published guidelines on the diagnosis and treatment of these injuries. OBJECTIVES: THE PRESENT SYSTEMATIC REVIEW WAS UNDERTAKEN TO ANSWER THE FOLLOWING QUESTIONS: what are the most commonly recommended diagnostic criteria for finger flexor pulley injury in rock climbers; and, based on the available evidence, what is the best diagnostic test for these injuries? METHODS: Four electronic databases were searched using specific key terms, with limits set for language and date. Two reviewers independently identified potentially relevant titles based on inclusion criteria. Inter-reviewer variability was assessed using the Kappa statistic. The scientific quality of articles was assessed using validated scales. RESULTS: Of the 93 articles identified, 29 were included in the present analysis. The inter-rater agreement for selection of potentially relevant titles was 88% (kappa=0.74). The most commonly cited diagnostic criterion for closed finger pulley injury was clinical bow-stringing of the flexor tendons over the volar aspect of the proximal interphalangeal joint. However, the best study of diagnostic accuracy for these injuries supports the use of dynamic ultrasound. CONCLUSIONS: Dynamic ultrasound is recommended for the diagnosis of closed finger pulley injuries in rock climbers. The prevailing notion that these injuries can be diagnosed by testing for clinical bowstringing is not supported by evidence. PMID- 19554142 TI - Atypical presentation of a glomus tumour: A case report. AB - A 46-year-old man presented with an extremely painful, 1 cm, mobile, nodular mass located on the medial side of his right elbow. Symptoms failed to respond to conservative treatment. Ultrasound and Doppler flow imaging revealed a well defined, round hypoechoic mass. Under local anesthesia, the mass was resected and the wound closed without complication. Final pathology diagnosed the lesion as a glomus tumour, solid type. Histology staining showed the tumour cells were positive for antibodies to vimentin and muscle actin. It is unusual for a glomus tumour to be located anterior to the medial epicondyle of the right arm. Hand surgeons most commonly encounter glomus tumours in the nailbeds of the fingers. PMID- 19554144 TI - What works, what doesn't. PMID- 19554143 TI - Femoral nerve palsy as a complication of anterior iliac crest bone harvest: Report of two cases and review of the literature. AB - There are many documented neurological complications of anterior iliac crest bone harvest. Until now, these have included injuries to the iliohypogastric, subcostal and lateral femoral cutaneous nerves. Femoral nerve palsy as a direct surgical complication of anterior iliac crest bone harvest has never been cited in any surgical literature, although it has been reported in deep pelvic and abdominal surgeries in which improper retraction and/or prolonged hyperextension of the hip may have caused a nerve compression syndrome. In addition, surgical patients on antithrombolytic therapy have experienced hemorrhage within the iliacus and iliopsoas muscles, resulting in hematoma and secondary femoral nerve compression. The classic motor and sensory deficits reported in femoral nerve palsies are reduced or absent patellar reflex, weak hip flexion, quadriceps muscle weakness, and anesthesia of the anterior thigh and medial aspect of the leg. Two cases of femoral nerve palsy with different etiologies are presented. PMID- 19554145 TI - Giant lipomas of the upper extremity. AB - Lipomas are slow-growing soft tissue tumours that rarely reach a size larger than 2 cm. Lesions larger than 5 cm, so-called giant lipomas, can occur anywhere in the body but are seldom found in the upper extremities. The authors present their experiences with eight patients having giant lipomas of the upper extremity. In addition, a review of the literature, and a discussion of the appropriate evaluation and management are included. PMID- 19554146 TI - Composite tissue allotransplantation of the face: Decision analysis model. AB - BACKGROUND: Facial composite tissue allotransplantation is a potential reconstructive option for severe facial disfigurement. The purpose of the present investigation was to use decision analysis modelling to ascertain the expected quality-adjusted life years (QALYs) gained with face transplantation (versus remaining in a disfigured state) in an effort to assist surgeons with the decision of whether to adopt this procedure. STUDY DESIGN: The probabilities of potential complications associated with facial allotransplantation were identified by a comprehensive review of kidney and hand transplant literature. A decision analysis tree illustrating possible health states for face allotransplantation was then constructed. Utilities were obtained from 30 participants, using the standard gamble and time trade-off measures. The utilities were then translated into QALYs, and the expected QALYs gained with transplantation were computed. RESULTS: Severe facial deformity was associated with an average of 7.34 QALYs. Allotransplantation of the face imparted an expected gain in QALYs of between 16.2 and 27.3 years. CONCLUSIONS: The current debate within the medical community surrounding facial composite tissue allotransplantation has centred on the issue of inducing a state of immunocompromise in a physically healthy individual for a non-life-saving procedure. However, the latter must be weighed against the potential social and psychological benefits that transplantation would confer. As demonstrated by a gain of 26.9 QALYs, participants' valuation of quality of life is notably greater for face transplantation with its side effects of immunosuppression than for a state of uncompromised physical health with severe facial disfigurement. PMID- 19554147 TI - Self-inflicted specific pattern burns in psychiatric patients. AB - Self-inflicted burns represent a major social and medical problem for society. Differences have been demonstrated between patients who attempt suicide and those who deliberately harm themselves without any intention of killing themselves. These self-inflicted injuries may resemble injuries that are intentionally inflicted by others and may require investigation by protective services. Little is known about these specific pattern burn injuries in psychiatric patients. PMID- 19554148 TI - Lawsuits against plastic surgeons: Does locale affect incidence of claims? AB - BACKGROUND: Physicians continue to practice in a very litigious environment. Some physicians try to mitigate their exposure to lawsuits by avoiding geographical locations known for their high incidence of medical malpractice claims. Not only are certain areas of the United States known to have a higher incidence of litigation, but it is also assumed that certain areas of the hospital incur a greater liability. There seems to be a medicolegal dogma suggesting a higher percentage of malpractice claims coming from patients seen in the emergency room (ER), as well as higher settlements for ER claims. OBJECTIVE: To determine if there is any validity to the dogma that a higher percentage of malpractice claims arise from the ER. METHODS: An analysis of common plastic surgery consults that result in malpractice claims was performed. The location where the basis for the lawsuit arose - the ER, office (clinic) or the operating room (OR) - was evaluated. The value of the indemnity paid and whether its value increased or decreased based on the location of the misadventure was evaluated. RESULTS: According to the data, which represented 60% of American physicians, there was a larger absolute number of malpractice claims arising from the OR, not the ER. However, the highest average indemnity was paid for cases involving amputations when the misadventure originated in the ER. CONCLUSIONS: The dogma that a greater percentage of lawsuits come from incidents arising in the ER is not supported. However, depending on the patient's injury and diagnosis, a lawsuit from the ER can be more costly than one from the OR. PMID- 19554149 TI - Pilomatricoma: experience of the hospital for sick children. AB - BACKGROUND: Pilomatricoma (calcifying epithelioma of Malherbe) is a common skin neoplasm in the pediatric population that is often misdiagnosed as other skin conditions or tumours. OBJECTIVES: The objective of the present retrospective study was to review the clinical and histopathological presentation of this neoplasm in children. METHODS: The records of the pathology department at The Hospital for Sick Children, Toronto, Ontario, were searched for all cases of pilomatricoma between 2001 and 2006. The records of these patients were reviewed to determine sex, age, location and size of the tumour, pathological features and recurrence rate. All patients underwent surgical excision of the lesions. RESULTS: A total of 93 lesions in 85 patients were identified. The median age was 8.7 years. Of the 85 patients diagnosed with pilomatricoma, 44 (52%) were female. In all cases, the initial presentation was an asymptomatic, slow growing, superficial hard mass with bluish discolouration. The most common sites of occurrence were the face (48%), neck (21%) and upper limbs (18%). The size of the surgical specimens collected ranged from 0.1 cm to 2.6 cm. The diagnosis was confirmed by histopathological examination in all cases. Ghost cells and basaloid cells were described in most of the cases (83%). There were no recurrences in this series. CONCLUSIONS: This entity should be considered with other benign or malignant conditions in the clinical differential diagnosis of solitary firm skin nodules, especially those on the face, neck and upper limbs. The diagnosis can generally be made by clinical examination. The treatment of choice is surgical excision, and the recurrence rate is very low. PMID- 19554150 TI - Angiomyxoma diagnosed in a man presenting for abdominal lipectomy. AB - Aggressive angiomyxoma is a rare tumour that is locally infiltrative but nonmetastasizing. It occurs nearly exclusively in adult women of childbearing age and almost always arises in the perineum and pelvic area. A case of angiomyxoma occurring in a middle-aged, morbidly obese man is reported. The clinical presentation was one of progressing scrotal edema and enlargement with subsequent development of scrotal abscesses, requiring several incisions and drainages as well as oral and intravenous antibiotics. His symptoms and body habitus left him in a significantly debilitated state, prompting him to seek treatment for his conditions. He initially presented for abdominal lipectomy. However, due to the chronic scrotal infections and enlargement, scrotectomy was recommended before any other surgical procedures. A scrotectomy was performed by the urology service. Pathological diagnosis of the excised tumour was an aggressive angiomyxoma. Given the rarity of this tumour, especially in males, the proper diagnosis and treatment of angiomyxoma is still being investigated. With this in mind, the physical signs and symptoms that characterize this condition are presented and treatment options are reviewed. PMID- 19554151 TI - Extraskeletal Ewing's sarcoma in a great toe of a young boy. AB - Extraskeletal Ewing's sarcoma (EES) is a rare, soft tissue, malignant neoplasm histologically similar to skeletal Ewing's sarcoma. It occurs mainly in adolescents and young adults, and affects extremities in 36% of cases and central locations (commonly paravertebral regions) in the remainder. The differential diagnosis includes other small, blue, round cell tumours. A clinical case of EES involving a great toe in a young boy is reported. EES diagnosis was confirmed by features of histological analysis and immunohistochemistry, and by the presence of the t(11;22) chromosomal translocation. PMID- 19554152 TI - Atypical fibroxanthoma in a young woman: An unusual case presentation. AB - Atypical fibroxanthoma (AFX) is an uncommon neoplasm, identified as a spindle cell tumour that is generally found in elderly patients on sun-exposed areas. The majority of cases of AFX are benign, and metastasis is a rare phenomenon. The first case in the literature of AFX is described in a young woman with no previous risk factors who presented with a three-month history of an enlarging nodule of the left nasal alar. Excision showed the lesion to be composed of hyperchromatic, pleomorphic, vacuolated spindle cells and multinucleated giant cells. The tumour cells stained positive for macrophage-histiocyte antigen alpha(1)-antitrypsin, neurokinin-1, CD68 and alpha(1)-antichymotrypsin. The present case report highlights the importance of correct diagnosis for AFX with adequate excision and by considering the histopathology and immunohisto-chemistry of its clinical differential diagnosis. PMID- 19554153 TI - Post-traumatic pseudolipoma of the forehead. AB - A forehead lipoma is a rare finding in a child, and one that penetrates the underlying layers of muscle and bone to attach to dura has not previously been reported. Two such cases, both in children who underwent uneventful deliveries aided by forceps, are presented. Both lesions were present at birth and, based on clinical findings, were originally thought to be dermoid cysts. Dermoid cysts could not be ruled out with computed tomography and magnetic resonance imaging. Histopathology identified fibrofatty tissue consistent with lipoma. Both lesions extended from the subcutaneous tissue through the frontalis muscle and frontal bone to the dura. Given these findings and the history of forceps delivery, the most likely diagnosis is post-traumatic pseudolipoma. This lesion should be considered in the differential diagnosis of congenital lesions of the forehead, particularly if there is a history of forceps delivery or other trauma to the area. PMID- 19554154 TI - Facial neurothekeoma in a 10-year-old child. AB - Neurothekeoma is a rare and benign tumour that occurs most frequently on the head and neck. It is classified as a nerve sheath tumour and epidemiologically occurs in young adults. It is rare to find these tumours occurring in children. The following report describes a case of neurothekeoma in a young girl, its treatment and a review of the literature. Specific histological analysis may reveal atypical and aggressive features, which, in turn, affect resection and reconstructive options. PMID- 19554155 TI - Office anesthesia for breast augmentation made easy. PMID- 19554157 TI - Doing surgery. PMID- 19554156 TI - Emergency finger fractures - An easy fix. PMID- 19554158 TI - A new technique of internal suture mastopexy for mild to moderate breast ptosis. AB - BACKGROUND: Current mastopexy techniques rely on incisions on the breast to correct ptosis. Trading a ptotic breast for a visibly scarred breast can be a difficult choice. OBJECTIVE AND METHODS: A technique of internal suture mastopexy that consists of plicating sutures placed in the superficial fascia of the breast from the deep surface is presented. The procedure leaves no scar on the breast and may be safer than other techniques when combining mastopexy with augmentation. RESULTS: The senior author has performed this procedure on over 120 patients, with a mean follow-up of two years. Patients and the surgeon have expressed satisfaction with the procedure. CONCLUSION: Based on this experience with over 120 patients, the authors believe that internal suture mastopexy is an effective alternative in selected patients. PMID- 19554159 TI - Minimal inframammary incision for breast augmentation. AB - The inframammary approach in breast augmentation, still the most popular technique among plastic surgeons, has always been hampered by the undesirable appearance of its scar. The present paper describes a modified approach to inframammary augmentation with saline-filled prostheses. This approach uses a very short incision, thus resulting in a much less noticeable scar. The surgical technique is easy to learn, simple to execute, does not necessitate any special equipment and gives consistent results. Decreasing the scar length to an absolute minimum ensures higher patient and surgeon satisfaction. PMID- 19554160 TI - Predicting breast reduction weight using the mass of breast ptosis. AB - BACKGROUND: The preoperative prediction of therapeutic breast reduction weights, to achieve both relief of breast weight symptoms and yet achieve excellent breast shape, remains a challenge. OBJECTIVES: To design a simple clinical method to preoperatively predict and quantify therapeutic breast reduction weights. METHODS: In 31 women who underwent therapeutic bilateral reduction mammaplasty, the mass of the hypertrophic breast hanging below the inframammary fold was preoperatively weighed and then compared with the mass of the reduction specimen. Thirty patients underwent breast reduction using a superomedial nipple-areolar pedicle. Postoperative breast weight-related symptoms and breast shape findings were then noted. Statistical analysis relied on mean, SD, sample size, Mann Whitney test for medians, Levene's test for variances and regression analysis. RESULTS: The average clinical follow-up was 160 days, with all patients achieving satisfactory breast size and shape from both the patient and surgeon's perspectives. All patients reported improvement of back pain, shoulder pain and lower neck pain. Two breasts developed delayed healing of the lateral skin flap, necessitating debridement and reclosure, followed by uneventful ongoing healing. There was no significant difference in preoperative ptotic breast mass and resectional breast mass (all P>0.05). CONCLUSIONS: Simple preoperative weighing of the ptotic portion of the hypertrophic breast can serve as a goal for the reduction weight, while creating pleasing breast proportions and improving breast weight-related symptoms. Preoperative quantification of the ptotic breast mass may guide the reduction technique and assist insurance precertification efforts. PMID- 19554161 TI - The combined plastic surgery/physical medicine and rehabilitation amputee clinic at the University of Western Ontario. AB - Since the autumn of 2001, a multidisciplinary plastic surgery (PS) and physical medicine and rehabilitation (PMR) outpatient amputee clinic has been in place at St Joseph's Health Centre/Parkwood Hospital in London, Ontario. To date, more than 140 new patients have been seen in combined consultations. The present paper reviews the demographics, interventions and outcomes of the patients seen between 2001 and 2005. The majority of primary PMR patients had problems that prevented optimal use of their lower extremity prostheses. These problems included nonhealing pressure ulcers, infections, painful neuromas, amputation stump shape abnormalities and fixed joint contractures. Most patients had surgical intervention. In terms of overall clinical success, 53% of the PMR patients and 77% of the PS patients had their problems resolved. The multidisciplinary collaboration in the PS/PMR outpatient amputee clinic provides a unique service that benefits upper and lower extremity amputees. PMID- 19554162 TI - Client satisfaction of hand therapy intervention: An evaluation of the effectiveness of therapy for clients recovered from complex regional pain syndrome. AB - Complex regional pain syndrome (CRPS) is a neuropathic pain condition that may develop following trauma to an extremity. Clients treated for CRPS at St Joseph's Health Care London - Hand and Upper Limb Centre, London, Ontario, were asked to evaluate their level of satisfaction with the treatment they had received by comparing their pain, functional status and emotional status before and after receiving therapy. The results indicated a high level of satisfaction among clients, attributable to the unique nature of the therapy program in use at this facility, where the occupational therapist works in close collaboration with the surgeon and pain specialists, and the therapy regimen is designed for each client individually according to his or her needs. The unique contribution of the present study to the body of clinical literature on CRPS is that it introduces a focus on client functionality and on client satisfaction with therapy received. PMID- 19554163 TI - Identification of mandibular fracture epidemiology in Canada: Enhancing injury prevention and patient evaluation. AB - BACKGROUND: Mandibular fractures can lead to significant functional and aesthetic sequelae if treated improperly. They may act as an indicator of concomitant trauma and are very demanding on the public health care system. Thus, knowledge of mandibular fracture epidemiology is critical to effective prevention, as well the establishment of accurate trauma evaluation protocols. OBJECTIVES: To identify the epidemiology of mandibular fractures treated at a level 1 Canadian trauma centre, clarify the pathogenesis of these epidemiological patterns and suggest potential targets for preventive efforts. METHODS: A retrospective review of all mandibular fracture patients presenting to the Montreal General Hospital between 1998 and 2003 was performed. Medical records and digitized radiographic imaging were used to collect patient demographics and injury data. RESULTS: The chart review identified 181 patients with 307 mandibular fractures. Fifty-two per cent of the fractures occurred in individuals 21 to 40 years of age, 78% of patients were male, and there was wide ethnic diversity. Sixty percent of patients had multiple mandibular fractures; 29% were symphyseal/parasymphyseal fractures, 25% were condylar fractures and 23% were angle fractures. Assault was the most common mechanism of injury, with 29% of fractures involving alcohol or illegal drug use. Thirty percent of patients had an associated facial fracture, and more than one-third had another major injury. CONCLUSIONS: The present epidemiological review reveals several potential prevention targets as well as significant trends. Further research into the impact of these preventive measures could more objectively identify their impact on mandibular trauma. PMID- 19554165 TI - A geometric method for nipple localization. AB - BACKGROUND: An important part of preoperative assessment in breast reduction surgery is to locate the site of the nipple-areola complex for the newly structured breast. Inappropriate location is difficult to correct secondarily. Traditional methods of nipple localization taught and practiced suggest the nipple to be located anterior to the inframammary fold. Trying to project this point on the anterior surface of the breast requires either large calipers or feeling the posteriorly placed finger on the anterior surface of a large breast. This certainly introduces some subjectivity to the calculation. OBJECTIVES: To introduce an easy and accurate method of nipple localization to reduce the learning curve for trainee surgeons. METHODS: Aesthetic placement of the nipples is at the lower angles of an equilateral or a short isosceles triangle on the chest with its apex at the sternal angle. This triangle can be thought of as two right-angled triangles with their Y-axis on the median plane. The base and vertical limb are measured, and the hypotenuse is calculated. The location of the lower angle is marked on the anterior surface of the breast and represents the new position of the nipple. RESULTS: Forty patients had nipple localization performed in the above-described manner, with satisfactory placement of the nipple-areola complex. CONCLUSIONS: The above technique introduces some objective measurements to the localization of the nipple in breast reduction surgery. It is easy to practice, and infuses confidence in trainees marking their initial breast reductions. PMID- 19554164 TI - Melanotic neuroectodermal tumour of infancy. AB - Melanotic neuroectodermal tumour of infancy is an extremely rare neoplasm arising in newborns and young children, typically involving the face or cranium. A case arising from the maxilla, requiring extensive resection with a near-total maxillectomy, is presented. A thorough review of the literature on this unusual tumour is provided, with emphasis on prognostic factors and appropriate treatment. PMID- 19554166 TI - Dupuytren's contracture following burn injury of the hand: A case report and review of literature. AB - In burn patients, scar contractures adjacent to or across the joints lead to disabling deformities. In Dupuytren's disease, the proliferative process involves the fascia of the palm and fingers, resulting in disabling flexion contractures of the fingers and the palm. A single insult involving the hand or even a more proximal injury may lead to Dupuytren's disease. PMID- 19554167 TI - Fat injections made quick and easy. PMID- 19554168 TI - Tendonitis in the upper extremity: Is it a useful or useless diagnosis? PMID- 19554169 TI - Predicting the results of rhinoplasty before surgery: Easy noses versus difficult noses. AB - A major problem for many rhinoplastic surgeons is the ability to predict, before surgery, the difficulty of the procedure (whether the rhinoplasties will be technically easy or technically difficult to perform) and the success rate of the result (whether the rhinoplasty will likely give good results or poor ones).The present paper outlines a systematic approach to nasal analysis, allowing the surgeon to consistently estimate, before surgery, the degree of technical difficulty of each rhinoplasty, as well as predicting its future result in terms of patient satisfaction. This preoperative evaluation is based on the analysis of the skin texture and the osteocartilagenous framework on lateral and frontal views. It allows for the nose to be classified as green (easy), yellow (moderate) or red (difficult), depending on two factors: the degree of surgical difficulty and the expected patient's satisfaction with the result.The essence of the present paper is to introduce a simple, systematic approach to assist the novice rhinoplastic surgeon to assess the complexity, the risks and the expected outcome of a rhinoplasty in the preoperative period, rather than postoperatively. PMID- 19554170 TI - A comparison of the superficial inferior epigastric artery flap and deep inferior epigastric perforator flap in postmastectomy reconstruction: A cost-effectiveness analysis. AB - PURPOSE: To perform a cost-effectiveness analysis comparing the superficial inferior epigastric artery (SIEA) and deep inferior epigastric perforator (DIEP) flaps in postmastectomy reconstruction. METHODS: A decision analytic model with seven clinically important health outcomes (health states) was used, incorporating the Ontario Ministry of Health's perspective. Direct medical costs were estimated from a university-based hospital. The utilities of each health state converted into quality-adjusted life years (QALYs) were obtained from previously published data. Health state probabilities were computed from a systematic literature review. Analyses yielded SIEA and DIEP expected costs and QALYs allowing calculation of the incremental cost-utility ratio (ICUR). One-way sensitivity analyses were conducted under five plausible scenarios, assessing result robustness. RESULTS: Five SIEA and 27 DIEP studies were identified. The baseline SIEA expected cost was slightly higher than that for the DIEP ($16,107 versus $16,095), with slightly higher QALYs (33.14 years versus 32.98 years), giving an ICUR of $77/QALY. Taking into account conversions from SIEA to DIEP, the ICUR increased to $4,480/QALY. Sensitivity analysis gave ICURs ranging from $2,614/QALY to 'dominant', all consistent with the adoption of the SIEA over the DIEP. CONCLUSION: The best available evidence suggests the SIEA is a cost effective procedure. However, given the high SIEA to DIEP conversion rates and small marginal differences in cost and effectiveness, the ICUR may be sensitive to minor changes in costs or QALYs. The 'truth' can only be obtained from a randomized, controlled trial comparing both techniques side by side, simultaneously capturing the costs of the competing interventions. PMID- 19554171 TI - Patient satisfaction following nipple reconstruction incorporating autologous costal cartilage. AB - BACKGROUND: Nipple-areolar reconstruction completes post-mastectomy breast reconstruction. Many techniques for nipple reconstruction have been described, and each has their advocates and critics. One of the frequent failings of most designs is loss of nipple projection with time. OBJECTIVES: To determine the effect of including autologous costal cartilage on patient satisfaction with their nipple reconstruction. METHODS: Sixty-eight patients were identified who had undergone fishtail flap nipple reconstruction following autologous free flap breast reconstruction between 1990 and 2004. Qualitative questionnaires, using Likert scales, were sent to each patient to specifically assess their satisfaction with their nipple reconstruction. RESULTS: Of 26 respondents (mean +/- SEM follow-up period 3.7+/-3.6 years), 13 had undergone nipple reconstruction incorporating costal cartilage banked at the time of initial breast reconstruction, and the other 13 had no cartilage in their nipple reconstructions. While both groups would opt for nipple reconstruction again, patients with cartilage grafts incorporated into their reconstructions had overall satisfaction ratings 1.92 grades higher on average (not significant, P=0.12) than those without. This difference increased to 3.2 grades when the satisfaction of the patient's partner was taken into account (P<0.05). Improved satisfaction corresponded to higher scores for volume, consistency, texture, and particularly for projection and contour of the nipple (P<0.05). Although nipple morphology changed over time, there was a trend toward improved stability in the cartilage group. CONCLUSIONS: Patient satisfaction with nipple reconstruction can be improved by incorporating costal cartilage beneath the skin flaps. Superior contour and projection are sustained over time. PMID- 19554172 TI - Post-traumatic osteomyelitis of the clavicle: A case report and review of literature. AB - Osteomyelitis of the clavicle is a rare form of infection occurring from hematogenous spread or trauma. This has been reported following head and neck surgery, and subclavian catheter placement. In traumatic cases, the management involves removal of bone fixation, debridement of the bone and coverage with a muscle flap. PMID- 19554173 TI - Extra-anatomic microvascular second toe transfer for creation of a helper hand - a long-term follow-up. AB - Mutilating hand injuries can significantly alter a patient's life. Return to work and daily activities remain the ultimate goal. A case report and long-term follow up of a patient who caught his hand in a generator fan at work is presented. He sustained an injury that left him with a wrist and metacarpal stump. When prostheses failed, an extra-anatomic second toe transfer was used to produce a functional distal extremity. This transfer allowed the patient to return to his job and was surprisingly durable, resulting in a useful helper hand for 15 years. PMID- 19554174 TI - Past the postauricular graft. PMID- 19554175 TI - Rules for practice in 2008. PMID- 19554176 TI - Evaluation of three-dimensional computed tomography processing for deep inferior epigastric perforator flap breast reconstruction. AB - BACKGROUND: The deep inferior epigastric perforator flap procedure has become a popular alternative for women who require breast reconstruction. One of the difficulties with this procedure is identifying perforator arteries large enough to ensure that the harvested tissue is well vascularized. Current techniques involve imaging the perforator arteries with computed tomography (CT) to produce a grid mapping the locations of the perforator arteries relative to the umbilicus. OBJECTIVES: To compare the time it takes to produce a map of the perforators using either two-dimensional (2D) or three-dimensional (3D) CT, and to see whether there is a benefit in using a 3D model. METHODS: Patient CT abdomen and pelvis scans were acquired from a GE 64-slice scanner. CT image processing was performed with the GE 3D Advantage Workstation v4.2 software. Maps of the perforators were generated both as 2D and 3D representations. Perforators within a region 5 cm rostral and 7 cm caudal to the umbilicus were measured and the times to perform these measurements using both 2D and 3D images were recorded by a stopwatch. RESULTS: Although the 3D method took longer than the 2D method (mean [+/- SD] time 1:51+/-0:35 min versus 1:08+/-0:16 min per perforator artery, respectively), producing a 3D image provides much more information than the 2D images alone. Additionally, an actual-sized 3D image can be printed out, removing the need to make measurements and producing a grid. CONCLUSIONS: Although it took less time to create a grid of the perforators using 2D axial CT scans, the 3D reconstruction of the abdomen allows the plastic surgeons to better visualize the patient's anatomy and has definite clinical utility. PMID- 19554177 TI - Management of proximal interphalangeal joint injuries. AB - Injuries to the proximal interphalangeal joint are common but frequently missed. They are often overtreated by prolonged immobilization, resulting in stiffness, which may be permanent. The purpose of the present article is to briefly review the relevant anatomy and biomechanics, present an approach to physical examination and diagnosis, and propose a practical clinical classification. The treatment of the most severe injury - the intra-articular fracture dislocation - is controversial. The various treatment options are discussed, based on personal experience and a review of the literature. A list of relevant references is presented. PMID- 19554178 TI - The 'bikini lip reduction': A detailed approach to hypertrophic lips. AB - Excessively large lips represent an occasional but significant challenge in aesthetic surgery. Previously described techniques focus largely on the simple excision of a strip of tissue to reduce the lips, without specific attention to the resultant lip contour or to the volume relationship between the lips. The present paper describes a new technique for lip reduction, called the 'bikini lip reduction'. This technique not only reduces the volume of the lips, but also restores an attractive labial contour, as well as an ideal volume relationship between the upper and lower lips. Because it is based on aesthetic analysis, this technique consistently yields both smaller and more aesthetically appealing lips. Simply stated, the bikini lip reduction consists of excision of a 'bikini top' (two cups and a middle strap) from the upper lip and a 'bikini bottom' (a triangle) from the lower lip. The aesthetic results and the patient satisfaction achieved through the bikini lip reduction technique have been very satisfactory. PMID- 19554179 TI - Management of fulminant dissecting cellulitis of the scalp in the pediatric population: Case report and literature review. AB - A case of fulminant dissecting cellulitis of the scalp in a fifteen-year-old African American male is reported. The presentation was refractory to standard medical treatment such that treatment required radical subgaleal excision of the entire hair-bearing scalp. Reconstruction was in the form of split-thickness skin grafting at the level of the pericranium following several days of vacuum assisted closure dressing to promote an acceptable wound bed for skin grafting and to ensure appropriate clearance of infection. Numerous nonsurgical modalities have been described for the treatment of dissecting cellulitis of the scalp, with surgical intervention reserved for patients refractory to medical treatment. The present paper reports a fulminant form of the disease in an atypical age of presentation, adolescence. The pathophysiology, etiology, natural history, complications and treatment options for dissecting cellulitis of the scalp are reviewed, and the authors suggest this method of treatment to be efficacious for severe presentations refractory to medical therapy. PMID- 19554180 TI - Treatment of congenital epulis (granular cell tumour) with excision and gingivoperiosteoplasty. AB - Congenital epulis, or granular cell tumour, is a rare, benign intraoral tumour present on the mucosa of the alveolar ridge, with the potential to interfere with respiration and feeding. Treatment involves surgical excision, and recurrences are rare. The management of the alveolar deficiency associated with this tumour has not previously been addressed. In the present report, a patient with a typical epulis, treated surgically with excision and extrapolation of Millard's gingivoperiosteoplasty technique to reconstruct the associated alveolar defect with restoration of alveolar ridge continuity, is presented. The differential diagnosis and complications of this tumour are discussed. PMID- 19554182 TI - Ulnar artery pseudoaneurysm in a patient with factor IX deficiency (hemophilia B). AB - The hypothenar hammer syndrome describes a constellation of symptoms resulting from repetitive trauma to the hypothenar eminence, often due to the use of the hand as a hammer. Sequelae of this syndrome include both true and false aneurysms, as well as thrombosis of the ulnar artery due to its vulnerability to blunt trauma as it exits Guyon's canal. Although this is a relatively well documented phenomenon, an extensive review of the literature revealed that no cases have been described involving a patient with hemophilia. The present case describes a 46-year-old farmer with factor IX deficiency (hemophilia B) presenting with a 5 cm x 7 cm pseudoaneurysm of the ulnar artery of the right hand. PMID- 19554181 TI - Giant submental lipoma: Case report and review of the literature. AB - Lipomas may be located in all parts of the body and may be confused clinically with other soft tissue masses. They infrequently occur in the head and neck. A large neck mass (greater than 10 cm) with a rapid growth rate should raise concerns about a possible malignancy. Failure to distinguish a liposarcoma from a lipoma may represent a medicolegal pitfall. Surgical excision of a lipoma is often used as the definitive treatment modality, and alternative treatments described for lipomas range from liposuction to steroid injections. In the present study, a 60-year-old man who presented with a rapidly enlarging submental mass is described. A 15 cm x 12 cm mass was successfully removed. The surgery produced excellent cosmetic results and no functional impairment. An integrated review of the literature regarding etiology, epidemiology, diagnostic and treatment modalities of submental lipomas follows. PMID- 19554183 TI - An apparent duplication of the mouth in a patient with midline dysraphism and a teratoid polyp: A 15-year postsurgical follow-up. AB - True median cleft of the upper and lower lip occurs very rarely. Complete or partial facial duplication is an extremely rare malformation. Approximately 26 cases have been reported since 1900, spanning a wide spectrum of clinical severity. Most cases appear to share a number of features, including cleft palate, duplication of the tongue, orbital hypertelorism and macrostomia. An unusual patient with midline clefts of the upper and lower lips with an apparent duplication of the mouth and a teratoid polyp, with fusion of the maxilla and mandible, is reported. The early management and surgical intervention is discussed. Furthermore, postoperative outcomes are reviewed as part of a 15-year postsurgical follow-up. PMID- 19554184 TI - An uncommon folliculosebaceous cystic hamartoma on the lower extremity. AB - BACKGROUND: Folliculosebaceous cystic hamartomas (FSCH) are a recently recognized cutaneous hamartoma comprised of follicular, sebaceous and mesenchymal elements. Only nine cases have been previously reported. The authors present a case of FSCH on the lower extremity in addition to a literature review. OBJECTIVE: To further characterize the clinical, histological and immunohistological features of FSCH. METHODS: The clinical, histological and immunohistological features of FSCH were evaluated and the literature reviewed. RESULTS: This lesion uncommonly presents as exophytic papules or nodules on the leg, and most commonly presents on the central part of the face. Clinical differential diagnoses included nevus, neurofibroma and adnexal tumour. The diagnostic microscopic features of infundibular cystic structures with attached sebaceous glands and characteristic stroma were present in all lesions. Many of the stromal spindle cells were CD34 positive. CONCLUSION: The FSCH is a recently recognized hamartoma comprised of follicular, sebaceous and mesenchymal elements. Although reported only rarely, its incidence is likely much higher. In the present patient, the FSCH was located in femoral region, which is very rare. PMID- 19554185 TI - Reply. PMID- 19554187 TI - That sinking feeling. PMID- 19554186 TI - Cortisone finger. PMID- 19554188 TI - The versatile rhomboid (Limberg) flap. AB - The rhomboid (Limberg) flap can be used to close defects almost anywhere on the body. It is versatile in that a random pattern flap can be raised from any one or all corners of the rhomboid. The defect is filled with tissue of the same thickness and colour, and with good vascularity. The present paper demonstrates the versatility of the rhomboid flap. PMID- 19554189 TI - Breast autoaugmentation. AB - A technique using a posteriorly based dermoglandular flap as an augmentation of the superior hemisphere of the breast combined with a periareolar mastopexy and vertical mastopexy is presented. The advantages of combining a periareolar mastopexy, in terms of reducing the length of the vertical scar and preventing areolar distortion, are explained. PMID- 19554191 TI - Forequarter amputation for malignant tumours of the upper extremity: Case report, techniques and indications. AB - Forequarter (interscapulothoracic) amputation is a major ablative surgical procedure that was originally described to manage traumatic injuries of the upper extremity. Currently, it is most commonly used in the treatment of malignant tumours of the arm. With the advent of limb-sparing techniques, primary forequarter amputation is performed less frequently, but remains a powerful surgical option in managing malignant tumours of the upper extremity; therefore, surgeons should be familiar with this procedure. A classic case report of forequarter amputation, with emphasis on indications and surgical techniques, is presented. PMID- 19554190 TI - Facial paralysis for the plastic surgeon. AB - Facial paralysis presents a significant and challenging reconstructive problem for plastic surgeons. An aesthetically pleasing and acceptable outcome requires not only good surgical skills and techniques, but also knowledge of facial nerve anatomy and an understanding of the causes of facial paralysis.The loss of the ability to move the face has both social and functional consequences for the patient. At the Facial Palsy Clinic in Edinburgh, Scotland, 22,954 patients were surveyed, and over 50% were found to have a considerable degree of psychological distress and social withdrawal as a consequence of their facial paralysis. Functionally, patients present with unilateral or bilateral loss of voluntary and nonvoluntary facial muscle movements. Signs and symptoms can include an asymmetric smile, synkinesis, epiphora or dry eye, abnormal blink, problems with speech articulation, drooling, hyperacusis, change in taste and facial pain.With respect to facial paralysis, surgeons tend to focus on the surgical, or 'hands on', aspect. However, it is believed that an understanding of the disease process is equally (if not more) important to a successful surgical outcome. The purpose of the present review is to describe the anatomy and diagnostic patterns of the facial nerve, and the epidemiology and common causes of facial paralysis, including clinical features and diagnosis. Treatment options for paralysis are vast, and may include nerve decompression, facial reanimation surgery and botulinum toxin injection, but these are beyond the scope of the present paper. PMID- 19554192 TI - Canadian Society of Plastic Surgeons Societe Canadienne des Chirurgiens Plasticiens: Abstracts presented at the 61st Annual Meeting / 61 Reunion annuelle May 29 - June 2, 2007, Banff, Alberta. PMID- 19554193 TI - Groupe pour L'Avancement de la Microchirurgie Canada (GAM): Abstracts presented at the 28th Annual Meeting / 28 Reunion annuelle May 29, 2007, Banff, Alberta. PMID- 19554194 TI - Particulate matter air pollution exposure: role in the development and exacerbation of chronic obstructive pulmonary disease. AB - Due to the rapid urbanization of the world population, a better understanding of the detrimental effects of exposure to urban air pollution on chronic lung disease is necessary. Strong epidemiological evidence suggests that exposure to particulate matter (PM) air pollution causes exacerbations of pre-existing lung conditions, such as, chronic obstructive pulmonary disease (COPD) resulting in increased morbidity and mortality. However, little is known whether a chronic, low-grade exposure to ambient PM can cause the development and progression of COPD. The deposition of PM in the respiratory tract depends predominantly on the size of the particles, with larger particles deposited in the upper and larger airways and smaller particles penetrating deep into the alveolar spaces. Ineffective clearance of this PM from the airways could cause particle retention in lung tissues, resulting in a chronic, low-grade inflammatory response that may be pathogenetically important in both the exacerbation, as well as, the progression of lung disease. This review focuses on the adverse effects of exposure to ambient PM air pollution on the exacerbation, progression, and development of COPD. PMID- 19554196 TI - Asthma patients prefer Respimat Soft Mist Inhaler to Turbuhaler. AB - Device satisfaction and preference are important patient-reported outcomes to consider when choosing inhaled therapy. A subset of adults (n = 153) with moderate or severe asthma participating in a randomized parallel-group, double dummy trial that compared the efficacy and safety of 12 weeks' treatment with budesonide delivered via Respimat Soft Mist Inhaler (SMI) (200 or 400 microg bd) or Turbuhaler dry powder inhaler (400 microg bd), completed a questionnaire on patient device preference and satisfaction (PASAPQ) as part of a psychometric validation. As the study used a double-dummy design to maintain blinding, patients used and assessed both devices, rating their satisfaction with, preference for, and willingness to continue using each device. The mean age of patients was 41 years, 69% were female and the mean duration of disease was 16 years. Total PASAPQ satisfaction scores were 85.5 and 76.9 for Respimat SMI and Turbuhaler respectively (p < 0.0001); 112 patients (74%) preferred Respimat SMI and 26 (17%) preferred Turbuhaler. Fourteen subjects (9%) indicated no preference for either inhaler. Willingness to continue using Respimat SMI was higher than that for Turbuhaler (mean scores: 80/100 and 62/100, respectively). Respimat SMI was preferred to Turbuhaler by adult asthma patients who used both devices in a clinical trial setting. PMID- 19554197 TI - Hormones and Borderline Personality Features. AB - Borderline personality is diagnosed in clinical settings three times more often in women than in men, and symptom severity in women appears sensitive to circulating sex steroid levels. In non-human mammals, prenatal hormones contribute to the development of sex-linked behavior and their responsiveness to postnatal hormones. Therefore, this study examined the hypothesis that prenatal hormones may influence the development of borderline personality traits by measuring a marker of perinatal androgen action, the 2D:4D ratio, and salivary hormone levels in 58 men and 52 women. Participants completed the Borderline Features Subscales (BOR) of the Personality Assessment Inventory, gender role questionnaires, and four sex-linked cognitive tasks. Digit ratios were a significant predictor of the affective component of borderline personality, such that in both sexes 2D:4D ratios suggestive of weaker perinatal androgen action contributed to greater borderline personality features overall and greater affective instability. In addition, women reporting greater affective instability showed larger changes in estradiol across the session, consistent with the influence of stress and emotional reactivity on hormonal function. These findings are consistent with an increasing body of research suggesting that hormonal factors associated with the expression of typical gender-linked behavior may also contribute to the expression of gender-linked maladaptive behavior. PMID- 19554198 TI - Conflicts of Interest in Scientific Research Related to Regulation or Litigation. AB - This article examines conflicts of interest in the context of scientific research related to regulation or litigation. The article defines conflicts of interest, considers how conflicts of interest can impact research, and discusses different strategies for dealing with conflicts of interest. While it is not realistic to expect that scientific research related to regulation or litigation will ever be free from conflicts of interest, society should consider taking some practical steps to minimize the impact of these conflicts, such as requiring full disclosure of information required for independent evaluation of research, prohibiting financial relationships between regulatory agencies and the companies they regulate, and banning payments to expert witnesses for specific research results, testimony or legal outcomes. PMID- 19554199 TI - Which is more important in bioimaging SIMS experiments-The sample preparation or the nature of the projectile? AB - Sample preparation is central to acquiring meaningful molecule-specific images with SIMS, especially when submicron lateral resolution is involved. The issue is to maintain the distribution of target molecules while attempting to introduce biological cells or tissue into the high vacuum environment of the mass spectrometer. Here we compare freeze-drying, freeze-etching, freeze-fracture and trehalose vitrification as possible strategies for these experiments. The results show that the prospects for successful imaging experiments are greatly improved with all of these methods when using cluster ion bombardment, particularly C(60) (+) ions, not only due to increased sensitivity of this projectiles, but also since it removes contamination overlayers without insult to the underlying chemistry. The emergence of 3-dimensional imaging capabilities also suggests that sample preparation should not perturb the 3-dimensional morphology of the cell, a situation not generally possible during freeze-drying. Hence, sample preparation and projectile type are strongly coupled parameters for bioimaging with mass spectrometry. PMID- 19554195 TI - Exacerbation rate, health status and mortality in COPD--a review of potential interventions. AB - COPD is prevalent in Western society and its incidence is rising in the developing world. Acute exacerbations of COPD, about 50% of which are unreported, lead to deterioration in quality of life and contribute significantly to disease burden. Quality of life deteriorates with time; thus, most of the health burden occurs in more severe disease. COPD severity and frequent and more severe exacerbations are all related to an increased risk of mortality. Inhaled corticosteroids (ICS) have similar effects on quality of life but ICS/long-acting bronchodilator combinations and the long-acting antimuscarinic tiotropium all improve health status and exacerbation rates and are likely to have an effect on mortality but perhaps only with prolonged use. Erythromycin has been shown to decrease the rate of COPD exacerbations. Pulmonary rehabilitation and regular physical activity are indicated in all severities of COPD and improve quality of life. Noninvasive ventilation is associated with improved quality of life. Long term oxygen therapy improves mortality but only in hypoxic COPD patients. The choice of an inhaler device is a key component of COPD therapy and this requires more attention from physicians than perhaps we are aware of. Disease management programs, characterized as they are by patient centeredness, improve quality of life and decrease hospitalization rates. Most outcomes in COPD can be modified by interventions and these are well tolerated and have acceptable safety profiles. PMID- 19554200 TI - Plasma Carotenoids and Biomarkers of Oxidative Stress in Patients with prior Head and Neck Cancer. AB - Diets high in fruits and vegetables are generally believed protective against several chronic diseases. One suggested mechanism is a reduction in oxidative stress. The carotenoids, nutrients found in colored fruits and vegetables, possess antioxidant properties in vitro, but their role in humans is less well documented. The aim of this cross-sectional study was to explore the relationships between the most abundant plasma carotenoids (alpha-carotene, beta carotene, lycopene, lutein, zeaxanthin and beta-cryptoxanthin), as well as grouped carotenoids (total xanthophylls, carotenes and carotenoids), and urinary excretion of the F(2)-isoprostanes (F(2)-IsoPs), stable and specific biomarkers of oxidative damage to lipids. Two F(2)-IsoP measures were utilized: total F(2) IsoPs and 8-iso-PGF(2alpha). The study population (N = 52) was drawn from a study among patients curatively treated for early-stage head and neck cancer. Unadjusted linear regression analyses revealed significant inverse associations between plasma lutein, total xanthophylls and both F(2)-IsoP measures at baseline. After control for potential confounders, all individual and grouped xanthophylls remained inversely associated with the F(2)-IsoP measures, but none of these associations achieved significance. The carotenes were not inversely associated with total F(2)-IsoPs or 8-iso-PGF(2a) concentrations. The finding of consistent inverse associations between individual and grouped xanthophylls, but not individual and grouped carotenes, and F(2)-IsoPs is intriguing and warrants further investigation. PMID- 19554201 TI - The effect of incident angle on the C(60) bombardment of molecular solids. AB - The effect of incident angle on the quality of SIMS molecular depth profiling using C(60) (+) was investigated. Cholesterol films of ~300 nm thickness on Si were employed as a model and were eroded using 40 keV C(60) (+) at an incident angle of 40 degrees and 73 degrees with respect to the surface normal. The erosion process was characterized by determining at each angle the relative amount of chemical damage, the total sputtering yield of cholesterol molecules, and the interface width between the film and the Si substrate. The results show that there is less molecule damage at an angle of incidence of 73 degrees and that the total sputtering yield is largest at an angle of incidence of 40 degrees . The measurements suggest reduced damage is not necessarily dependent upon enhanced yields and that depositing the incident energy nearer the surface by using glancing angles is most important. The interface width parameter supports this idea by indicating that at the 73 degrees incident angle, C(60) (+) produces a smaller altered layer depth. Overall, the results show that 73 degrees incidence is the better angle for molecular depth profiling using 40 keV C(60) (+). PMID- 19554202 TI - Iridium(I)-Catalyzed Regio- and Enantioselective Allylic Amidation. AB - Ir(I)-catalyzed intermolecular allylic amidation of ethyl allylic carbonates with soft nitrogen nucleophiles under completely "salt-free" conditions is described. A combination of [Ir(COD)Cl](2), a chiral phosphoramidite ligand L*, and DBU as a base in THF effects the reaction. The reaction appears to be quite general, accommodating a wide variety of R-groups and soft nitrogen nucleophiles, and proceeds with excellent regio- and enantioselectivities to afford the branched N protected allylic amines. The developed reaction was conveniently utilized in the asymmetric synthesis of Boc protected alpha- and beta-amino acids as well as (-) cytoxazone. PMID- 19554203 TI - New Technologies for Diagnosing Pediatric Tumors Expert Opinion on Medical Diagnostics. AB - BACKGROUND: The completion of Human Genome Project (HGP) has paved the way for novel, more detailed and accurate molecular diagnostic classification of cancer. With the information from the HGP, cancers can be categorized not only on the morphology or limited immunohistological markers, but according to their "molecular fingerprints" such as gene expression profiles. Technologies detecting these signatures have been developed to simultaneously measure multiple genes or proteins in one assay with high sensitivity and specificity. OBJECTIVE: To evaluate potential innovative novel methods of diagnosis and prognosis in pediatric cancers. METHODS: We selected a variety of promising new diagnostic technologies utilizing molecular signatures which harness the results from HGP including DNA microarray, bead-based detection system, multiplexed RT-PCR, MesoScale Discovery (MSD), and isotope-coded affinity tag (ICAT), as well as their applications in biomarker discovery for pediatric tumors. Label-free detection technologies and the obstacles for taking these new diagnostic technologies from the bench to the bedside are also discussed. CONCLUSION: The use of molecular signatures is gaining acceptance in clinical practice. However, technical challenges need to be addressed before incorporating these new technologies into current diagnostic and prognostic schema. PMID- 19554204 TI - Determination of Flavonoids and Resveratrol in Wine by Turbulent-Flow Chromatography-LC-MS. AB - Turbulent-flow chromatography (TFC) on-line coupled to liquid chromatography mass spectrometry (LC-MS) is used to determine flavonoids and resveratrol in different types of wines. A fully automated system was developed in which 10 mL of sample (diluted wine) was passed over a TFC column, after which the retained analytes were separated by reversed-phase LC and detected by negative ion mode atmospheric pressure chemical ionization (APCI) MS. The method proved to be fast, non laborious, robust and sensitive. The feasibility of the method was tested on several red, white and rose wines. Quantitation of resveratrol was possible using the standard addition procedure. Red wine showed the highest amount of resveratrol (4 mg L(-1)), while rose and white wine contained concentrations which were about ten fold lower. PMID- 19554205 TI - Enantioselective Reduction of Prochiral Ketones using Spiroborate Esters as Catalysts. AB - Novel spiroborate esters derived nonracemic 1,2-aminoalcohols and ethylene glycol are reported as highly effective catalysts for the asymmetric borane reduction of a variety of prochiral ketones with borane-dimethyl sulfide complex at room temperature. Optically active alcohols were obtained in excellent chemical yields using 0.1 to 10 mol % of catalysts with up to 99% ee. PMID- 19554206 TI - Photopatternable Polymeric Membranes for Optical Oxygen Sensors. AB - A new class of optical oxygen sensor that can be photopatternable by traditional UV lithography is presented. They are fabricated using photopatternable spin-on silicone (polydimethyl-siloxane, PDMS) with oxygen sensitive luminescent dyes. It has a good adhesion property and can be applied on glass or on photopolymer (SU 8) without any additional surface treatments. The optimum mixture composition for patternable oxygen sensitive membranes is investigated and its optical properties are characterized. Proof-of-concepts for two applications, intensity-based oxygen sensing with SU-8 based structure and self-calibration fluidic oxygen sensor, are described. These photopatternable optical membranes will find many applications wherever small patterns of oxygen sensitive membranes are required. PMID- 19554207 TI - Selection of Protein Crystal Forms Facilitated by Polymer-Induced Heteronucleation. AB - Crystallization of biological macromolecules as high quality single crystals is critical for determining their structure and facilitates the rational design of drugs. Because macromolecules often crystallize in multiple phases that have unique diffraction properties, the selective production of phases is desirable. Furthermore, determining multiple structures allows for a greater understanding of the relationship between crystal packing and conformation. With the aim of exploiting the polymer-induced heteronucleation approach to selectively nucleate multiple macromolecule crystal forms, hen egg white lysozyme (HEWL) was chosen as a model. Selective phase production was achieved under conditions that, in the absence of added heteronuclei, result in crystallization of a single crystal form. Moreover, nucleation rate, which in turn affects the size and quality of HEWL crystals, was controlled by various polymer surfaces. Thus, the polymer induced heteronucleation approach provides an additional diversity element which can be easily implemented to complement standard crystal growth techniques for the selective production of high quality protein crystals. PMID- 19554208 TI - All Smiles are Not Created Equal: Morphology and Timing of Smiles Perceived as Amused, Polite, and Embarrassed/Nervous. AB - We investigated the correspondence between perceived meanings of smiles and their morphological and dynamic characteristics. Morphological characteristics included co-activation of Orbicularis oculi (AU 6), smile controls, mouth opening, amplitude, and asymmetry of amplitude. Dynamic characteristics included duration, onset and offset velocity, asymmetry of velocity, and head movements. Smile characteristics were measured using the Facial Action Coding System (Ekman, Friesen, & Hager, 2002) and Automated Facial Image Analysis (Cohn & Kanade, 2007). Observers judged 122 smiles as amused, embarrassed, nervous, polite, or other. Fifty-three smiles met criteria for classification as perceived amused, embarrassed/nervous, or polite. In comparison with perceived polite, perceived amused more often included AU 6, open mouth, smile controls, larger amplitude, larger maximum onset and offset velocity, and longer duration. In comparison with perceived embarrassed/nervous, perceived amused more often included AU 6, lower maximum offset velocity, and smaller forward head pitch. In comparison with perceived polite, perceived embarrassed more often included mouth opening and smile controls, larger amplitude, and greater forward head pitch. Occurrence of the AU 6 in perceived embarrassed/nervous and polite smiles questions the assumption that AU 6 with a smile is sufficient to communicate felt enjoyment. By comparing three perceptually distinct types of smiles, we found that perceived smile meanings were related to specific variation in smile morphological and dynamic characteristics. PMID- 19554209 TI - Processing distinct linguistic information types in working memory in aphasia. AB - BACKGROUND: Recent investigations have suggested that adults with aphasia present with a working memory deficit that may contribute to their language-processing difficulties. Working memory capacity has been conceptualised as a single "resource" pool for attentional, linguistic, and other executive processing alternatively, it has been suggested that there may be separate working memory abilities for different types of linguistic information. A challenge in this line of research is developing an appropriate measure of working memory ability in adults with aphasia. One candidate measure of working memory ability that may be appropriate for this population is the n-back task. By manipulating stimulus type, the n-back task may be appropriate for tapping linguistic-specific working memory abilities. AIMS: The purposes of this study were (a) to measure working memory ability in adults with aphasia for processing specific types of linguistic information, and (b) to examine whether a relationship exists between participants' performance on working memory and auditory comprehension measures. METHOD #ENTITYSTARTX00026; PROCEDURES: Nine adults with aphasia participated in the study. Participants completed three n-back tasks, each tapping different types of linguistic information. They included the PhonoBack (phonological level), SemBack (semantic level), and SynBack (syntactic level). For all tasks, two n-back levels were administered: a 1-back and 2-back. Each level contained 20 target items; accuracy was recorded by stimulus presentation software. The Subject-relative, Object-relative, Active, Passive Test of Syntactic Complexity (SOAP) was the syntactic sentence comprehension task administered to all participants. OUTCOMES #ENTITYSTARTX00026; RESULTS: Participants' performance declined as n-back task difficulty increased. Overall, participants performed better on the SemBack than PhonoBack and SynBack tasks, but the differences were not statistically significant. Finally, participants who performed poorly on the SynBack also had more difficulty comprehending syntactically complex sentence structures (i.e., passive & object-relative sentences). CONCLUSIONS: Results indicate that working memory ability for different types of linguistic information can be measured in adults with aphasia. Further, our results add to the growing literature that favours separate working memory abilities for different types of linguistic information view. PMID- 19554210 TI - A Culturally Informed Model of Academic Well-Being for Latino Youth: The Importance of Discriminatory Experiences and Social Support. AB - This study tested a culturally informed model of academic well-being for 278 Latino youth. We examined detrimental effects of discriminatory experiences and protective effects of social support on self-reported academic outcomes. Models specified main and buffering effects of social support and compared contributions of support provided by parents, school, and peers. Data indicated that discrimination was associated with lower academic well-being, social support buffered effects of discrimination on academic well-being, and parental support was most predictive of greater academic well-being. Combined sources of social support were more important than any one source alone. Implications for culturally specified research, preventive interventions, and practitioners are discussed. PMID- 19554211 TI - Autoimmune retinopathy in systemic lupus erythematosus: histopathologic features. AB - The ocular pathology of autoimmune retinopathy is demonstrated in a 62-year-old female patient with systemic lupus erythematosus (SLE) who presented with typical clinical autoimmune retinopathy. Macroscopically, there were multiple depigmented lesions in the peripheral retina and choroid and scattered pigmentary bone spickling at the equator and periphery. Microscopically, there were generalized loss of photoreceptors and thinning of the outer plexiform layer. Many peripheral retinal vessels were sclerotic and occluded, some surrounded by pigment granules and RPE cells. Cobblestone degeneration was prominent in the periphery. Macrophages were seen in the retina, particularly in areas of photoreceptor degeneration. Rare, scattered T- lymphocytes were present in the retina and choroid, while B-cells were notably absent. The optic nerve showed loss of axons and thickened septae. Serum autoantibodies against normal retinal nuclei were detected. These pathological changes represent both known SLE-associated ocular complications as well as possible features of autoimmune retinopathy secondary to SLE. PMID- 19554212 TI - The Use of Polarization Filters to Detect the Edge of the Descemet's Stripping Automated Endothelial Keratoplasty(DSAEK) Graft. AB - PURPOSE: To report the technique of combining the two different polarization filters to detect the flap edge of the corneoscleral tissue before trephining the Descemet's stripping automated endothelial keratoplasty (DSAEK) tissue. METHODS: A human DSAEK donor tissue was prepared with mechanical microkeratome and the tissue on the cutting block was brought under the microscope. The liner, circular, or the combination of these two polarization filters was placed between the tissue and the microscope. The tissue images were taken with digital camera under either of 3 settings. RESULTS: The combination of circular and linear polarization filters enabled us to recognize the edge of the flap more easily than others. CONCLUSION: This simple system with polarization filters was effective in clear visualization of the flap edge during DSAEK tissue preparation. These features may significantly enhance safety of various surgical procedures, in addition to DSAEK tissue preparation. PMID- 19554213 TI - Cross-clade recognition of HIV-1 CAp24 by CD4+ T cells in HIV-1-infected individuals in Burkina Faso and Germany. AB - The presence of antigen-specific cellular immune responses may be an indicator of long-term asymptomatic HIV-1-disease. The detection of cellular immune responses to infection with different subtypes of HIV-1 may be hampered by genetic differences of immunodominant antigens such as the capsid protein CAp24. In Nouna, Burkina Faso, HIV-1 circulating recombinant forms CRF02_AG and CRF06_cpx are the 2 major strains detectable in HIV-1-infected individuals, while subtype B strains prevail in Europe and North America. Amino acid sequences of CAp24 were assessed in blood samples from 10 HIV-1-infected patients in Nouna, Burkina Faso. Production of interferon-gamma (IFN-gamma) in peripheral blood CD4(+) lymphocytes in response to recombinant HIV-1 proteins derived from clade B (including CAp24(NL4-3)) was measured using a modified flow-cytometry-based whole blood short term activation assay (FASTimmune, BDBiosciences). IFN-gamma production following stimulation with a whole length CAp24 protein derived from clade B (CAp24(NL4-3)) was additionally quantified in comparison to a CAp24 protein derived from CRF02_AG (CAp24(BD6-15)) in 16 HIV-1-infected patients in Heidelberg, Germany. Amino acid sequence identity of CAp24 obtained from patients in Nouna ranged between 86 and 89% when compared to the clade B CAp24(NL4-3) consensus sequence, between 90 and 95% when compared to the circulating recombinant form CRF06_CPX consensus sequence, and between 92 and 96% when compared to the CAp24(BD6-15) consensus sequence. Significant numbers of HIV-1 specific CD4(+) lymphocytes producing IFN-gamma were detected in 4 of 10 HIV-1 infected patients. In 7 of 16 patients in Heidelberg, recombinant CAp24(BD6-15) stimulated IFN-gamma-production in CD4(+) lymphocytes to a similar extent as the clade B-derived CAp24(NL4-3). Thus, antigen-specific CD4(+) lymphocytes from both West African and European patients infected with different strains of HIV-1 show relevant cross-clade recognition of HIV-1 CAp24 in a flow-cytometry-based whole blood short term activation assay. PMID- 19554214 TI - RNA detection and subtype C assessment of HIV-1 in infants with diarrhea in Ethiopia. AB - In the absence of chemoprophylaxis, HIV-1 transmission occurs in 13-42% of infants born to HIV-1 positive mothers. All exposed infants acquire maternal HIV 1 antibodies that persist for up to 15 months, thereby hampering diagnosis. In resource limited settings, clinical symptoms are the indices of established infection against validated laboratory-based markers. Here we enrolled 1200 children hospitalized for diarrheal and other illnesses. 20-25% of those tested, aged 15 months or younger, were found to be HIV-1-seropositive. Where sufficient plasma was available, HIV-1 RNA detection was performed using a subtype insensitive assay, with 71.1% of seropositive infants presenting with diarrhea showing positive. From sub-typing analysis, we identified that viruses of the C' sub-cluster were predominated amongst infants. Although this study may overestimate the HIV-1 frequency through testing symptomatic infants, diarrhea can be seen as a useful marker indicating HIV-1 infection in infants less than 15 months old. PMID- 19554215 TI - Problematics of Time and Timing in the Longitudinal Study of Human Development: Theoretical and Methodological Issues. AB - Studying human development involves describing, explaining, and optimizing intraindividual change and interindividual differences in such change and, as such, requires longitudinal research. The selection of the appropriate type of longitudinal design requires selecting the option that best addresses the theoretical questions asked about developmental process and the use of appropriate statistical procedures to best exploit data derived from theory predicated longitudinal research. This paper focuses on several interrelated problematics involving the treatment of time and the timing of observations that developmental scientists face in creating theory-design fit and in charting in change-sensitive ways developmental processes across life. We discuss ways in which these problematics may be addressed to advance theory-predicated understanding of the role of time in processes of individual development. PMID- 19554216 TI - Household Context and Subjective Well-Being Among the Oldest Old in China. AB - This article investigates the importance of household context to subjective well being among the oldest old (aged 80 years and older) in China. Using data from the Chinese Longitudinal Healthy Longevity Survey, the authors find that living arrangements have strong implications for elderly emotional health. First, living alone is associated with lower subjective well-being. Second, coresidence with immediate family (spouse or children) is associated with positive subjective well being. Third, compared to living with a son, the traditionally dominant type of living arrangement, coresidence with a daughter appears positively linked to the emotional health of the oldest old. Results highlight the importance of family and cultural context to subjective well-being of the oldest old. They also suggest that the gendered nature of caregiving merits further attention in China and other patrilineal societies. PMID- 19554217 TI - Judging near and distant virtue and vice. AB - We propose that people judge immoral acts as more offensive and moral acts as more virtuous when the acts are psychologically distant than near. This is because people construe more distant situations in terms of moral principles, rather than attenuating situation-specific considerations. Results of four studies support these predictions. Study 1 shows that more temporally distant transgressions (e.g., eating one's dead dog) are construed in terms of moral principles rather than contextual information. Studies 2 and 3 further show that morally offensive actions are judged more severely when imagined from a more distant temporal (Study 2) or social (Study 3) perspective. Finally, Study 4 shows that moral acts (e.g., adopting a disabled child) are judged more positively from temporal distance. The findings suggest that people more readily apply their moral principles to distant rather than proximal behaviors. PMID- 19554218 TI - The Importance of Clinicians Reviewing CT Scans in Suspected Lacrimal Gland Disease Causing Eyelid Swelling, Even if Radiologists Previously Interpreted them as Normal. AB - PURPOSE: To highlight the importance of deliberate evaluation of the lacrimal gland during routine orbital imaging by both radiologists and clinicians, which may avoid delays in diagnosis of eyelid swelling related to lacrimal gland disease. METHODS: We present four cases referred to a tertiary ophthalmic plastic and orbital centre for assessment of chronic upper eyelid swelling of unknown aetiology. All four cases had been investigated with orbital CT imaging, reported to be normal by consultant radiologists. RESULTS: Careful review of existing imaging provided valuable information concerning the lacrimal gland and helped identify a pathological cause in all four cases. CONCLUSIONS: Lacrimal gland disease can be missed radiologically without evaluation of coronal as well as standard axial orbital imaging. Clear communication with the radiologist as well as personally reviewing scans can avoid such pitfalls. PMID- 19554220 TI - Subconjunctival Air Leakage After Descemet's Stripping Automated Endothelial Keratoplasty(DSAEK) in a Post-Trabeculectomy Eye. AB - PURPOSE: To report a case of subconjunctival air leakage from the anterior chamber (AC) into a trabeculectomy bleb after Descemet's stripping automated endothelial keratoplasty (DSAEK). METHODS: An 89 year-old woman with a previous history of primary open angle glaucoma in her left eye and previous trabeculectomy with mitomycin C had DSAEK on the patient's left eye in order to treat her endothelial disease. During the DSAEK procedure, air was injected into the AC to aid in graft adherence. RESULTS: The day after the surgery, subconjunctival air leakage from the AC into a trabeculectomy bleb was observed. CONCLUSIONS: Although our patient did not have any complications from this leak, there exists the potential for hypotony, bleb-related infections, and dislocations of the DSAEK graft. Given the potential consequences, these patients should be monitored closely. PMID- 19554219 TI - Protein Z Plasma Levels are Not Elevated in Patients with Non-Arteritic Anterior Ischemic Optic Neuropathy. AB - BACKGROUND: Protein Z is a glycoprotein that acts as a co-factor for the inhibition of activated coagulation factor X. Protein Z circulating in abnormal levels has been associated with increased risk for acute ischemic events. Non arteritic Anterior Ischemic Optic Neuropathy (N-AION) is caused by acute ischemic infarction of the optic nerve head, supplied by the posterior ciliary arteries. OBJECTIVES: The aim was to investigate whether there is an association between N AION and plasma protein Z levels. PATIENTS AND METHODS: Twenty-six cases of confirmed N-AION and fifty-two controls were included in the study group. Protein Z was estimated in thawed citrate plasma on both N-AION cases and controls by an enzyme immunoassay. The imprecision of the estimation was satisfactory (CV = 4, 6%). RESULTS: The controls' protein Z values distributed within a range 340 to 4200 ng/ml (median = 1420, mean = 1673, SD = 1040 ng/ml). Patients' protein Z values distributed within a range 420 to 3600 ng/ml (median = 1030, mean = 1520, SD = 939 ng/ml). There was no statistical difference between the two distributions (Independent t-test, p=0.529). CONCLUSION: In our study, protein Z levels are not implicated in the pathogenesis of non-arteritic anterior ischemic optic neuropathy (N-AION). PMID- 19554221 TI - Repeatability and reproducibility of macular thickness measurements using fourier domain optical coherence tomography. AB - AIM: To evaluate repeatability and reproducibility of macular thickness measurements in visually normal eyes using the Topcon 3D OCT-1000. METHODS: Phase 1 investigated scan repeatability, the effect of age and pupil dilation. Two groups (6 younger and 6 older participants) had one eye scanned 5 times pre and post- dilation by 1 operator. Phase 2 investigated between-operator, within and between-visit reproducibility. 10 participants had 1 un-dilated eye scanned 3 times on 2 separate visits by 2 operators. RESULTS: PHASE 1: No significant difference existed between repeat scans (p=0.75) and no significant difference was found pre- and post-dilation (p=0.54). In the younger group variation was low (95% limits +/- 3.62microm) and comparable across all retinal regions. The older group demonstrated greater variation (95% limits +/- 7.6microm). PHASE 2: For a given retinal location, 95% confidence limits for within-operator, within-visit reproducibility was 5.16microm. This value increased to 5.56microm for the same operator over two visits and to 6.18microm for two operators over two visits. CONCLUSION: A high level repeatability, close to 6microm, of macular thickness measurement is possible using the 3D OCT- 1000. Measured differences in macular thickness between successive visits that exceed 6microm in pre-presbyopic individuals are therefore likely to reflect actual structural change. OCT measures are more variable in older individuals and it is advisable to take a series of scans so that outliers can be more easily identified. PMID- 19554222 TI - Frequency Doubling Technology vs Standard Automated Perimetry in Ocular Hypertensive Patients. AB - BACKGROUND: Frequency doubling technology (FDT) perimetry measures contrast sensitivity. The magnocellular component of ganglion cells in human retina is isolated as a whole by the FDT stimulus. The aim of this study is to investigate the role of Humphrey Matrix threshold testing in the detection of early functional retinal impairment in ocular hypertensive patients compared to standard automated perimetry (SAP). METHODOLOGY: Forty hypertensive patients were enrolled in this longitudinal observational clinical study. Functional testing included randomly Humphrey Matrix perimetry and white-on-white Humphrey visual field perimetry. Ibopamine test was performed in all forty patients. The cut-off of 3 mmHg was considered positive for this provocative test. RESULTS: Out of 40 patients, we included 21 in ibopamine positive group and 19 in ibopamine negative group. These two groups are sex- and age-matched. In ibopamine positive group the mean increase in IOP is 4.6 mmHg (ranging from 3 to 10 mmHg). Statistics showed that correlation between FDT and SAP was statistically significant in ibopamine negative group and not statistically significant in ibopamine positive group. Only one patient, coming from IBO + group, converted from ocular hypertension to glaucoma. All the other subjects remained stable in both groups without any therapy and visual field abnormalities. CONCLUSIONS: FDT showed to be more sensitive and specific than SAP mostly in detection of early visual field impairment in ocular hypertensive patients. PMID- 19554223 TI - The now popular and 'fashionable' carpal tunnel syndrome - revisited. PMID- 19554224 TI - A cost-utility analysis of open and endoscopic carpal tunnel release. AB - BACKGROUND: Open carpal tunnel release (OCTR) is the standard procedure for the surgical treatment of carpal tunnel syndrome. With the advent of minimally invasive surgery, endoscopic carpal tunnel release (ECTR) was introduced. OBJECTIVE: To use a decision analytical model to compare ECTR with OCTR in an economic evaluation. METHODS: Direct medical costs were obtained from a Canadian university hospital. Utility values obtained from experts, presented with carpal tunnel syndrome outcome health states, were transformed into quality-adjusted life years (QALYs). The probabilities of the health states associated with both techniques were obtained from the literature. RESULTS: The incremental cost utility ratio (ICUR) was $124,311.32/QALY gained, providing strong evidence to reject ECTR when ECTR is performed in the main operating room and OCTR is performed in the day surgery unit. A one-way sensitivity analysis in the present study demonstrated that when both OCTR and ECTR are performed in day surgery unit, the ICUR falls in the 'win-win' quadrant, making ECTR both more effective and less costly than OCTR. If the scar tenderness probability is decreased in the ECTR group in a second one-way sensitivity analysis, the ICUR decreases to $100,621.91/QALY gained, providing evidence to reject ECTR. If the reflex sympathetic dystrophy probability is increased in the ECTR group in a third one way sensitivity analysis, the ICUR increases to $202,657.88/QALY gained, providing strong evidence to reject ECTR. CONCLUSIONS: There is still uncertainty associated with the costs and effectiveness of ECTR and OCTR. To obtain a definitive answer as to whether the ECTR is more effective than the OCTR, it is necessary to perform a large, randomized, controlled trial in which the utilities and resource use are measured prospectively. PMID- 19554225 TI - The approach to frostbite in Turkey: A retrospective study. AB - Increased participation in outdoor activities and an epidemic of homelessness have caused the incidence of cold injuries in the civilian population of Turkey to rise dramatically during the past 20 years. Knowledge of treatment is crucial for emergency physicians in rural and urban areas. Recent developments have significantly advanced the understanding of the pathophysiology of hypothermic and frostbite injuries. The authors undertook a retrospective review of frostbite cases in the East Anatolia region of Turkey. The mean altitude of East Anatolia is 1600 m, and temperatures may be -35 degrees C at night and -18 degrees C in the morning in the cities and surrounding villages. Winter is prolonged and harsh, and freezing is a normal condition in this region. The socioeconomic level in this region is lower than other regions of Turkey. During the winter, roads may be closed for as long as five months, and most villages in East Anatolia have no medical units. Erzurum, one of the coldest cities in East Anatolia, is the only city with advanced medical hospitals in this region. Three hospitals in Erzurum were included in the present study: Ataturk University Medical Faculty, Erzurum Numune Hospital and Erzurum SSK Hospital. Frostbite case records and their properties are presented. PMID- 19554226 TI - Sterile versus nonsterile clean dressings. AB - BACKGROUND: Many patients cannot afford sterile dressings. In St John, New Brunswick, clean dressings have been used instead of sterile dressings for years, with no apparent ill effects. No previous studies have compared the sterility and cost of clean versus sterile dressing materials. OBJECTIVES: THE GOALS OF THE PRESENT STUDY WERE TO ANSWER THE FOLLOWING QUESTIONS: how much more sterile are sterile dressings than clean dressings; and how much does this extra sterility cost? METHODS: Sterility and cost of sterile gauze, panty liners, sanitary napkins, diapers and Coban tape (3M, USA) were compared. Samples, 2 cm x 2 cm in size, were cut out of each material under aseptic conditions, and delivered to the microbiology laboratory in sterile urine containers. The samples were then cultured and organisms were identified using conventional means. RESULTS: The cost for one month, using one 20 cm x 5 cm wound dressing daily, was calculated and compared with panty liners ($2.43), sanitary napkins ($5.55), diapers ($9.39) and Coban tape ($0.66), which were much cheaper than sterile dressings ($16.50). How sterile were the dressings? None of the 20 sanitary napkins grew bacteria, one of the 20 panty liners grew bacteria (coagulase-negative Staphylococcus), two of 20 sterile dressings grew bacteria (one coagulase-negative Staphylococcus and one nonhemolytic Streptococcus), 15 of 20 diapers grew bacteria (all bacillus) and two of five Coban rolls grew bacteria (one bacillus and one coagulase negative Staphylococcus). CONCLUSION: The panty liners, sanitary napkins and Coban tape studied were cheaper than, and had a comparible sterility with, the sterile gauze examined. PMID- 19554227 TI - The effectiveness of sodium tetradecyl sulfate in the treatment of wrist ganglia. AB - BACKGROUND: Ganglia are the most common benign soft tissue tumours of the hand. Although benign, a significant number of patients with wrist ganglia consult with their general practitioners, and ultimately the hand surgeon, complaining of pain. A great number of patients are concerned about the cosmetic appearance, and an equally significant number genuinely believe that the ganglion is a cancer. There are several management options resulting in varying degrees of success. These include observation only, surgical excision, aspiration only, aspiration with injection of methylprednisolone, and aspiration with injection of methylprednisolone and hyaluronidase. OBJECTIVE: The main objective was to investigate the effectiveness and safety of sodium tetradecyl sulfate as a sclerosing agent after aspiration of wrist ganglia. METHODS: Initial data were collected prospectively during a period of 48 months. Following this, patients were sent a postal survey at least two years after they had received treatment to access the levels of recurrence and persistent complications. RESULTS: In the short term, 90% of the patients achieved complete resolution after one episode of aspiration and injection. However, there was only a 65% cure rate after two to five years. Complications were few and not significant. CONCLUSION: Sodium tetradecyl sulfate is an effective sclerosing agent after aspiration of wrist ganglia with an excellent short-term efficacy and a long-term cure rate comparable to that of surgical excision. PMID- 19554228 TI - Cutaneous furuncular myiasis: Human infestation by the botfly. AB - Dermatobia hominis, the botfly, is indigenous to Central and South America. Its usual host is a mammal, often a horse or cow. Cutaneous furuncular myiasis, human infestation by the botfly, has rarely been reported. Symptoms of infestation include a locally painful, firm furuncular lesion, often with a centrally located pore. Due to their infrequent occurrence, these lesions are often misdiagnosed as cellulitis, leishmaniasis, furunculosis, staphylococcal boil, insect bite or sebaceous cyst - conditions with similar presentations. The present case reiterates the need to think of 'zebras' when hearing 'hoof beats' that may have originated in a different land. PMID- 19554229 TI - Isolated primary osteoma cutis of the head: Case report. AB - Osteoma cutis is a rare disease in which there is formation of bony tissue in the skin that causes deformities. The etiology remains unknown and its treatment is controversial. A rare case of primary osteoma cutis in the face and scalp, which was not associated with any syndrome, is described. The patient was treated with surgery and topical retinoic acid. The retinoic acid treatment resulted in an improvement of the frontal area, and stabilized the disease over a two-year follow-up period. Surgical resection was a simple treatment with a quick recovery, minimal scarring and no local recurrence. The patient was followed for two years and presented a satisfactory result.The treatment of osteoma cutis is quite variable, and surgery is the most frequently reported treatment. However, a combination of clinical and surgical treatments seems to be an efficient way to manage these patients. PMID- 19554230 TI - Pyoderma gangrenosum following reduction mammaplasty. AB - The failure of a postoperative local infection to resolve after appropriate antibiotic therapy should prompt consideration of other diagnoses. Reported here is a case of pyoderma gangrenosum, a rare necrotizing disorder, occurring after reduction mammaplasty. The clinical presentation was one of progressive wound deterioration with associated erythema and intense pain. After failure of antibiotic therapy and local wound care, tissue biopsy of the enlarging wound edge confirmed the diagnosis, which then responded rapidly to systemic steroid treatment. Given that the treatment for pyoderma gangrenosum is at odds with the standard treatment for an infection (steroids versus antibiotics), differentiating between the two diagnoses is vital to providing resolution of the process and limiting any untoward scarring resulting from the advancing open wounds that can develop. With this in mind, the physical signs and symptoms that characterize this condition and thus allow early diagnosis are presented, and treatment options discussed. PMID- 19554231 TI - Constriction band syndrome occurring in the setting of in vitro fertilization and advanced maternal age. AB - The debate as to the pathogenesis of constriction band syndrome began with Hippocrates and continues today. The exogenous theory attributes the condition to entanglement of the fetus in the amniotic remnants following premature rupture of the amnion, which is in contrast to the endogenous, or genetic, mechanism.A case of constriction band syndrome in the setting of in vitro fertilization, where the child was genetically unrelated to the birth mother, is presented. Constriction band syndrome has been reported following amniocentesis and chorionic villus sampling, but it has not heretofore been presented in the setting of in vitro fertilization. In addition, the present case presents an opportunity to separate maternal from genetic factors and, possibly, shed some light on the etiology of the condition. PMID- 19554232 TI - Congenital giant nevocellular nevus of the back with deep extension to the fat and fascia. AB - The presence of deep pigmentation in a giant congenital nevus makes its complete removal very difficult and should be taken into consideration when planning for treatment. A clinical case in which the pigmented lesion was found to extend deep into the muscle fascia is described. The lesion was surgically removed with an acceptable functional and aesthetic result through the utilization of tissue expansion. The development of new surgical techniques and the expansion of laser technology will possibly provide new treatment options in the future. Emphasis should be placed on aesthetics and function, because the excision based only on oncological anticipation is no longer valid. The presence of deep pigmentation makes certain treatment options less effective than surgical ablation. PMID- 19554233 TI - Intraneural lipoma of the ulnar nerve: A case report and review of literature. AB - Intraneural lipomas of the peripheral nerve or its cutaneous branches are rare benign tumours. These slow-growing tumours present as asymptomatic swelling, and diagnosis is usually made at the time of exploration. In most cases, these tumours can be enucleated without damage to the nerve fibres. PMID- 19554234 TI - Evacuation of hematomas using liposuction technology: Technique and literature review. AB - Established nonexpanding hematomas can be successfully treated with minimal morbidity using standard liposucstion techniques at the bedside or in an outpatient setting under local anesthesia. The authors presents a series of eight patients and discuss current concepts of dealing with this common and distressing surgical complication. PMID- 19554235 TI - A handy measuring device. PMID- 19554238 TI - Synthesis and stereochemical determination of the spirastrellolides. AB - This Highlight summarises synthetic efforts towards the marine sponge-derived antimitotic agent spirastrellolide A and its congeners, including the first total synthesis by Paterson and co-workers. PMID- 19554239 TI - Bioinformatic perspectives on NRPS/PKS megasynthases: advances and challenges. AB - The increased understanding of both fundamental principles and mechanistic variations of NRPS/PKS megasynthases along with the unprecedented availability of microbial sequences has inspired a number of in silico studies of both enzyme families. The insights that can be extracted from these analyses go far beyond a rough classification of data and have turned bioinformatics into a frontier field of natural products research. As databases are flooded with NRPS/PKS gene sequence of microbial genomes and metagenomes, increasingly reliable structural prediction methods can help to uncover hidden treasures. Already, phylogenetic analyses have revealed that NRPS/PKS pathways should not simply be regarded as enzyme complexes, specifically evolved to product a selected natural product. Rather, they represent a collection of genetic opinions, allowing biosynthetic pathways to be shuffled in a process of perpetual chemical innovations and pathways diversification in nature can give impulses for specificities, protein interactions and genetic engineering of libraries of novel peptides and polyketides. The successful translation of the knowledge obtained from bioinformatic dissection of NRPS/PKS megasynthases into new techniques for drug discovery and design remain challenges for the future. PMID- 19554240 TI - Blue flower color development by anthocyanins: from chemical structure to cell physiology. AB - Blue flower colors are primarily due to anthocyanin, a flavonoid pigment. Anthocyanin itself is purple in neutral aqueous solutions, ans its color is very unstable and quickly fades. Therefore, the mechanism of blue color development in living flower petals is one of the most intriguing problems in natural product chemistry. Much progress has been made in understanding blue flower coloration since the comprehensive review by Goto and Kondo in 1991. This review focuses on the advances in the last 15 years, and cites 149 references. PMID- 19554241 TI - Natural stilbenes: an overview. AB - Natural stilbenes have been a hot research topic due to their intricate structures and diverse biological activities. Although their molecular backbone consists only of 1,2-diphenylethylene units, stilbenes show an enormous diversity with regard to the different units present, the degree of polymerisation, and the pattern of oligomer construction. From January 1995 to the end of 2008, more than 400 new naturally occurring stilbenes were isolated and identified, and this review focuses on their structural diversity, distribution, and bioactivity. PMID- 19554242 TI - The Daphniphyllum alkaloids. AB - About two hundred Daphniphyllum alkaloids have so far been isolated from thirteen species of the genus Daphniphyllum. This comprehensive review summarizes all the research into the isolation and structure elucidation of these alkaloids reported since investigations began in 1996. The structures of the Daphniphyllum alkaloids are classified based on six Daphniphyllum alkaloids (daphniphylline, secodaphniphylline, yuzurimine, daphnilactone A, daphnilactone B and yuzurine); these are also a number of newly found skeletons. The biosynthetic pathways, total syntheses, and bioactivities of the Daphniphyllum alkaloids are also described. PMID- 19554245 TI - Suggested excess of occupational cancers in Norwegian offshore workers: preliminary results from the Cancer Registry Offshore Cohort. AB - OBJECTIVE: The aim of this communication was to report the overall incidence of cancer in a cohort of male Norwegian offshore oil workers. METHODS: The Offshore Cohort was comprised of >25,000 men who were employed at installations in the North Sea in the period 1965-1999, and who responded to a questionnaire that included work history offshore, other occupational experience, education, leisure time activities, and lifestyle factors. Calculating standardized incidence ratios (SIR), we compared the number of prospective incident cancers diagnosed between 1999 and 2005 with those expected for age-, gender- and period-specific rates in the general Norwegian population. RESULTS: The overall cancer incidence did not differ from that of the reference population [SIR=1.0, 95% -confidence interval (95% CI) 1.0-1.1, N=695]. There were indications of excess risks of acute myeloid leukemia (SIR=2.0, 95% CI 1.0-3.7) and cancer of the pleura (SIR=2.2, 95% CI 0.9 4.6). No data on occupational history was used in these preliminary analyses. CONCLUSIONS: The cohort was relatively young and an extended observation period would be important for in-depth analyses. The suggested excess of leukemia and cancer of the pleura may be linked to occupational exposure during employment offshore; this issue needs to be addressed in further studies. PMID- 19554244 TI - Impact of work-related factors, lifestyle, and work ability on sickness absence among Dutch construction workers. AB - OBJECTIVE: The objective of this study was to evaluate the relative contribution of individual characteristics, lifestyle factors, work-related risk factors, and work ability on the occurrence of short (<2 weeks), moderate (2-12 weeks), and long (>12 weeks) durations of sickness absence. METHODS: Altogether 5867 Dutch construction workers with complete sick leave registration were followed from the day of their medical examination in 2005 until the end of 2006. The main outcome of the study was the duration of sickness absence, as registered by an occupational health service. Independent variables consisted of individual characteristics, lifestyle factors, work-related factors, and the work ability index. We used Poisson regression analyses with repeated occurrence of sick leave to calculate rate ratios (RR) and 95% confidence intervals of independent variables for the three categories of sick leave duration. RESULTS: Predictors for sick leave lasting 2-12 weeks and >12 weeks were: older age, obesity, smoking, manual materials handling, lack of job control, lung restriction, and a less than excellent work ability. For most predictors, higher RR values were observed with a longer duration of sickness absence. Obesity, smoking, manual materials handling, and lack of job control remained important risk factors for moderate and long durations of sick leave after adjusting for the strong effects of work ability on sickness absence. The highest population-attributable fractions were observed for: age over 50 years (18%), manual materials handling (20%), and good (18%), moderate (28%), and poor (2%) work ability. CONCLUSION: This study suggests that a variety of preventive measures targeted at smoking, obesity, physical load, psychosocial work factors as well as work ability will contribute to a reduction in the occurrence of sick leave. PMID- 19554248 TI - Oxygen and guanine-cytosine profiles in marine environments. AB - One of the historic debates in molecular evolution concerns the strong variation in the genomic guanine-cytosine (GC) content of prokaryotes, which ranges from approximately 20-75%: Is this factor selectively neutral, or is it the result of natural selection? In a previous article published by our group, we showed that inside well-defined taxonomic groups of prokaryotes, strictly aerobic organisms tend to display higher genomic GC levels than strictly anaerobic species. In the present study, we examined the GC content of fragments of DNA obtained from microbial communities along a well-defined environmental gradient: a 4,000-m vertical profile in the North Pacific subtropical gyre. The patterns of GC distribution might be associated with oxygen concentrations in the seawater column. These results give further support to the link between a physiologic trait (aerobic respiration) and genomic GC content. PMID- 19554247 TI - Aplysia cys-loop glutamate-gated chloride channels reveal convergent evolution of ligand specificity. AB - Among the members of the superfamily of cys-loop ligand-gated ion channels (LGICs) are receptors distinguished by the presence of two cys-loops in the ligand-binding domain, for example, the glycine receptor. Such receptors have thus far been cloned only from vertebrates and from ecdysozoa (arthropods and nematodes). We have now cloned and expressed two 2-cys-loop receptors from Aplysia californica, a lophotrocozoan, and have shown that they form homomeric glutamate receptors. We have also built up a database including the two receptors cloned here, previously cloned vertebrate and ecdysozoan 2-cys-loop receptors taken from GenBank, and the same type of receptors obtained by a search of recently cloned genomes, including two non-vertebrate chordates, an echinoderm, a crustacean, an annelid, and another mollusk. We subjected these receptors to phylogenetic analysis, alone and in combination with GABA-A receptors from the same phyla and from a recently cloned cnidarian. The phylogenetic analysis revealed the presence of two independent clades of glutamate receptors: one from lophotrocozoa and other from ecdysozoa, and suggests that the ancestors of the current 2-cys-loop receptor types diverged from the GABA-A receptors and from each other before the bilateria-cnidaria split. Finally, combining the results from the phylogenetic analysis with those obtained from an analysis of the 2-cys loop receptors in light of recently published hypotheses concerning the glycine binding pocket, we predict that glycine receptors are not exclusively a vertebrate-receptor type. PMID- 19554246 TI - Femoral bone mineral density reflects histologically determined cortical bone volume in hemodialysis patients. AB - We evaluated the associations between dual energy X-ray absorptiometry (DXA) and histologically determined cancellous and cortical bone volume by controlling for vascular calcifications and demographic variables in hemodialysis (HD) patients. Femoral bone mineral density (f-BMD) was associated with cortical porosity. INTRODUCTION: Assessment of bone mass in chronic kidney disease patients is of clinical importance because of the association between low bone volume, fractures, and vascular calcifications. DXA is used for noninvasive assessment of bone mass whereby vertebral results reflect mainly cancellous bone and femoral results reflect mainly cortical bone. Bone histology allows direct measurements of cancellous and cortical bone volume. The present study evaluates the association between DXA and histologically determined cancellous and cortical bone volumes in HD patients. METHODS: In 38 HD patients, DXA was performed for assessment of bone mass, anterior iliac crest bone biopsies for bone volume, and multislice computed tomography for vascular calcifications. RESULTS: While lumbar bone mineral density (l-BMD) by DXA was not associated with histologically measured cancellous bone volume, coronary Agatson score showed a borderline statistically significant association (P = 0.055). When controlled for age and dialysis duration, f-BMD by DXA was associated with cortical porosity determined by histology (P = 0.005). CONCLUSIONS: The usefulness of l-BMD for predicting bone volume is limited most probably because of interference by soft tissue calcifications. In contrast, f-BMD shows significant association with cortical porosity. PMID- 19554249 TI - Subarcuate canal and artery: a case report. AB - Variations in the course of the subarcuate artery (SAA) and of its related funnel are infrequent and asymptomatic. We present a case of a 15-year-old girl with a preverbal severe bilateral hearing loss and a subarcuate canal with an unusual course and atypical correlation with the adjacent anatomical structures, particularly with the lateral semicircular canal and the facial nerve. The variation proposed in this case, not previously reported in the literature consulted, can have important implications during middle ear surgical procedures, particularly for the retro-facial tympanoplasty approach: in fact otologic surgeons should consider any kind of variation of the SAA in surgical planning to avoid unexpected hemorrhages. The CT-scan, particularly HRCT, is a very useful tool for the study of intrapetrous vessels and their funnels, and should be considered mandatory in the case of a posterior approach to the tympanic cavity. PMID- 19554250 TI - Extensor carpi radialis brevis origin, nerve supply and its role in lateral epicondylitis. AB - Lateral epicondylitis (LE) or tennis elbow has been the subject of concern during the last 60 years, but the pathogenesis of the LE remains unclear. The LE can be due to the tendinogenic, articular or neurogenic reasons. Numerous theories have been put fourth in the recent past, out of which one of the most popular theories is that the condition results from repeated contraction of the wrist extensor muscles, especially the extensor carpi radialis brevis (ECRB) which may compress the posterior branch of the radial nerve (PBRN) at the elbow during pronation. We studied 72 upper limbs (36 formalin-fixed cadaver) for the origin, nerve supply and the course of PBRN in relation to the ECRB as one of the goal for the present study. The possible presence of an arch of the ECRB around the PBRN was also observed and recorded. The nerve to ECRB was a branch from the radial nerve in 11 cases (15.2%); from the PBRN in 36 cases (50%) and from the superficial branch of the radial nerve in 25 cases (34.7%), respectively. The ECRB had a tendinous arch in 21 cases (29.1%); a muscular arch in 8 (11.1%) cases and the arch was absent in 43 cases (59.7%). When the ECRB had a tendinous or muscular arch around the PBRN, it may compress the same and this condition may worsen during the repeated supination and pronation as observed in tennis and cricket players. The presence of such tendinous or muscular arch should be considered by orthopedicians and neurosurgeons, while releasing the PBRN during LE surgery. PMID- 19554252 TI - The right vertebral artery arising as a branch of the right internal carotid artery: report of a rare case. AB - Atypical origins of the vertebral artery (VA) are rare anatomical findings. We present an extremely rare arrangement of the cerebral circulation in a 61-year old male patient detected on magnetic resonance angiography, where the right VA arose as a direct branch of the cervical internal carotid artery (ICA). This likely reflects the failure of the pro-atlantal artery to regress during the third to fourth week of embryological life. Although the effect of the variant on symptomology is debateable, knowledge of the potential for the VA to arise as a branch of the ICA is pertinent to radiologists and surgeons operating on the great vessels and their branches, particularly vascular surgeons undertaking carotid endarterectomy. PMID- 19554253 TI - [Diagnosis and treatment of femural neck fractures from the beginning to operative treatment: a historical overview]. AB - Operative stabilisation of femural neck fractures is a routine procedure nowadays and is usually performed without delay. Treatment by osteosynthesis or endoprosthesis allows immediate mobilisation with at least partial weight bearing.Immobilisation for months, associated with high mortality as practiced in former times, is nearly forgotten. We present a review on 430 years of diagnosis and treatment of these fractures since the first description by Ambroise Pare. In fact, it took 150 years to recognise the different locations of various hip fractures; at the time, fractures close to the caput were felt to have no chance of healing. Impacted fractures of the femural neck were first described 250 years after the first publication by A. Pare.This article also presents an overview on the development of various treatments to stabilise hip fractures: closed reduction and repositioning by internal rotation of the external rotated leg, positioning of the leg on the double inclined splint, different types of extension treatment and finally early operative procedures, when asepsis was established and fractures could be imaged by x-rays. PMID- 19554251 TI - Diameters of normal thoracic vascular structures in pediatric patients. AB - PURPOSE: To determine the normal range of aortic and pulmonary artery diameters on chest CT, and to search a constant ratio when the diameters of thoracic vascular structures are compared with an internal reference. METHODS: Contrast enhanced chest CT scans of 133 pediatric patients were retrospectively evaluated. Diameters of ascending and descending aorta, main pulmonary artery, right and left pulmonary arteries and a constant thoracic vertebra were measured. The mean ratios of thoracic vascular diameters to the diameter of the thoracic vertebra were calculated. RESULTS: There was a positive correlation between the age of the patients and vascular diameters. The mean ratios of vascular diameters to the diameter of thoracic vertebra, ranged from 1.1 for the ascending aorta to 0.70 for the right and left pulmonary arteries, were consistent. CONCLUSIONS: Diameters of thoracic vascular structures increase with age. The consistent vertebral to vessel ratios can be useful in evaluation of chest CT of pediatric patients. PMID- 19554254 TI - Loss of capture late after right ventricular pacing lead revision: what is the mechanism? PMID- 19554255 TI - Drug-eluting stent in malignant biliary obstruction. AB - INTRODUCTION: In unresectable malignant bile duct obstruction, endoscopic stent insertion is the treatment of choice. However, the current stent allows only mechanical palliation of the obstruction, and has no anti-tumor effect. Currently, in the vascular field, the drug-eluting stent (DES) is very highly favored. MATERIAL AND METHODS: The requirements for a DES in a non-vascular tract, such as the bile duct, are far different from those of a DES to be used in the vascular tract. The non-vascular DES must suppress tumor proliferation as well as mucosal hyperplasia. For example, the non-vascular stent might be covered with a membrane that gradually releases a chemo-agent. We do not have much experience with DES in the bile duct. Nonetheless, we are continuously testing many anti-tumor agents in animal and human studies. CONCLUSION: We expect and hope DES will work effectively for tumor cells in diverse ways and, more importantly, will prolong stent patency and the patients' survival periods. But considerable investigation and a clinical study of DES will be required to achieve these goals. PMID- 19554256 TI - Central sleep apnoea in Rett syndrome. AB - Breathing disturbances in Rett syndrome were reported almost entirely during wakefulness, with normal respiration during sleep. We studied a case of a proven MECP2 mutation in a girl, whose videopolygraphic and polysomnographic monitoring suggested the evidence of central apnoeas not only during awake, but also during sleep. Apart from prevalent awake respiratory dysfunction, central apnoeas in Rett syndrome may be also present during sleep. PMID- 19554257 TI - Transcriptional coactivator EDF-1 is required for PPARgamma-stimulated adipogenesis. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) is essential for adipogenesis. Since EDF-1 is a cofactor of PPARgamma, we investigated the molecular cross-talk between EDF-1 and PPARgamma in adipogenesis. While EDF-1 was not modulated during differentiation of 3T3-L1 cells, it co-immunoprecipitated with PPARgamma. Silencing EDF-1 by shRNAs inhibited the differentiation in adipocytes of 3T3-L1 cells, as detected by the staining of intracellular triglycerides and the expression of the PPARgamma target gene aP2. Accordingly, we found that anti-EDF-1 shRNAs decreased ligand dependent activation of PPARgamma in 3T3-L1 transiently transfected with a vector expressing luciferase under the control of a PPARgamma responsive consensus. To rule out that this inhibition is due to the concomitant downregulation of PPARgamma levels, we overexpressed PPARgamma in 3T3-L1 silencing EDF-1 and found a decrease of ligand dependent activation of PPARgamma, in spite of the high amounts of PPARgamma. These results demonstrate that EDF-1 is required for PPARgamma transcriptional activation during 3T3-L1 differentiation. PMID- 19554258 TI - Display of proteins on Bacillus subtilis endospores. AB - The targeting and anchoring of heterologous proteins and peptides to the outer surface of bacteriophages and cells is becoming increasingly important, and has been employed as a tool for fundamental and applied research in microbiology, molecular biology, vaccinology, and biotechnology. Less known are endospores or spores produced by some Gram-positive species. Spores of Bacillus subtilis are surrounded by a spore coat on their outside, and a few proteins have been identified being located on the outside layer and have been successfully used to immobilize antigens and some other proteins and enzymes. The major advantage of spores over the other published systems is their synthesis within the cytoplasm of the bacterial cell. Therefore, any heterologous protein to be anchored on the outside does not have to cross any membrane. Furthermore, spores are extremely resistant against high temperature, irradiation and many chemicals, and can be stored for many years at room temperature. PMID- 19554259 TI - Glucose binds to the insulin receptor affecting the mutual affinity of insulin and its receptor. AB - Insulin activity is sensitive to glucose concentration but the mechanisms are still unclear. An unexamined possibility is that the insulin receptor (IR) is sensitive to glucose concentration. We demonstrate here that insulin-like peptides derived from the IR bind glucose at low millimolar, and cytochalasin B at low micromolar, concentrations; several insulin-like IR peptides bind insulin at nanomolar Kd; and this binding is antagonized by increasing glucose concentrations. In addition, glucose and cytochalasin B bind to IR isolated from rat liver and increasing glucose decreases insulin binding to this IR preparation. The presence of GLUT 1 in our IR preparation suggests the possibility of additional glucose-mediated allosteric control. We propose a model in which glucose binds to insulin, the IR, and GLUT; insulin binds to the IR; and the IR binds to GLUT. This set of interactions produces an integrated system of insulin-dependent interactions that is highly sensitive to glucose concentration. PMID- 19554261 TI - Kank proteins: structure, functions and diseases. AB - The Kank family of proteins, Kank1-Kank4, are characterized by their unique structure, coiled-coil motifs in the N-terminal region, and ankyrin-repeats in the C-terminal region, with an additional motif, the KN motif, at the N-terminus. Kank1 was obtained by positional cloning of a tumor suppressor gene in renal cell carcinoma, while the other members were found by homology search. The family is involved in the regulation of actin polymerization and cell motility through signaling pathways containing PI3K/Akt and/or unidentified modulators/effectors. Their relationship to diseases such as cancer, and to neuronal and developmental disorders, will be an important subject of future study. PMID- 19554260 TI - Microbial thermosensors. AB - Temperature is among the most important of the parameters that free-living microbes monitor. Microbial physiology needs to be readjusted in response to sudden temperature changes. When the ambient temperature rises or drops to potentially harmful levels, cells mount protective stress responses--so-called heat or cold shock responses, respectively. Pathogenic microorganisms often respond to a temperature of around 37 degrees C by inducing virulence gene expression. There are two main ways in which temperature can be measured. Often, the consequences of a sudden temperature shift are detected. Such indirect signals are known to be the accumulation of denatured proteins (heat shock) or stalled ribosomes (cold shock). However, this article focuses solely on direct thermosensors. Since the conformation of virtually every biomolecule is susceptible to temperature changes, primary sensors include DNA, RNA, proteins and lipids. PMID- 19554262 TI - Nelfinavir induces mitochondria protection by ERK1/2-mediated mcl-1 stabilization that can be overcome by sorafenib. AB - The HIV protease inhibitor nelfinavir is an investigational drug for cancer treatment. We have previously demonstrated induction of apoptosis by nelfinavir even in chemo-resistant ovarian cancer cells. In contrast to the pro-apoptotic effect of nelfinavir on human cancer cells, we noticed a significant upregulation of the anti-apoptotic mitochondrial membrane protein mcl-1 by nelfinavir, resulting in a mitochondria-independent induction of apoptosis. Upregulation of mcl-1 was associated with enhanced phosphorylation of both mcl-1 and of ERK1/2 (extracellular signal-regulated kinases 1/2). ERK1/2 enhanced stability of mcl-1 protein expression by serine-163 phosphorylation. The combination of nelfinavir with sorafenib, a clinically applied inhibitor of the RAS/RAF/ERK1/2 pathway, inhibited nelfinavir-induced ERK1/2 activation and mcl-1 protein upregulation. Further, the combination of nelfinavir with sorafenib induced mitochondrial membrane potential disruption and resulted in an improved activity of nelfinavir on ovarian cancer cells. Thus, a combination of these two investigational anti cancer drugs could be of interest especially because of their unique mechanism of apoptosis induction even in otherwise chemo-resistant human cancer cells. PMID- 19554263 TI - Primary, non-exophytic, optic nerve germ cell tumors. AB - Tumors of the optic chiasm are relatively uncommon and usually associated with phakomatoses such as neurofibromatosis. Even more rare is the presentation of a primary, non-exophytic, isolated optic chiasm germ cell tumor (GCT). These tumors have imaging characteristics nearly indistinguishable from optic chiasmatic gliomas (OCGs). Herein we describe two cases of young men who presented with similar findings of progressive, painless visual loss and hypothalamic-pituitary adrenal axis dysfunction including diabetes insipidus. Brain imaging was non diagnostic and suggestive of an OCG. Pathology demonstrated GCTs in each case highlighting the importance of biopsy confirmation of the diagnosis. Both patients underwent a pterional craniotomy and sub-frontal approach to the optic chiasm. The chiasm was diffusely enlarged and discolored in each case without evidence of sellar, suprasellar or perichiasmatic pathology. Pathology demonstrated a malignant mixed GCT in the first patient and a germinoma in the second. This case series highlights the importance of tissue biopsy for patients with progressive symptoms from optic chiasm tumors. Furthermore, this is the first report of a primary, non-exophytic malignant mixed GCT. As the treatment regimens differ widely between optic chiasm GCTs and chiasm gliomas, tissue diagnosis is important. PMID- 19554264 TI - Fragment-based drug discovery. PMID- 19554266 TI - Transcriptional profiles during the differentiation and maturation of monocyte derived dendritic cells, analyzed using focused microarrays. AB - Dendritic cells (DC) are professional antigen-presenting cells capable of initiating primary immune responses. They have been intensively studied and are used in both basic immunology research and clinical immunotherapy. However, the genetic pathways leading to DC differentiation and maturation remain poorly understood. Using focused microarrays with oligonucletotide probes for 120 genes encoding co-stimulatory molecules, chemokines, chemokine receptors, cytokines, cytokine receptors, TLRs, and several other related molecules, we analyzed the kinetics of gene expression for the overall differentiation process of monocytes into mature DC. In parallel, we compared the transcriptional profiles in DC maturation in the presence of LPS, TNF-alpha or trimeric CD40L. We found similar transcriptional profiles for early immature DC and immature DC, respectively generated by culturing monocytes with GM-CSF and IL-4 for three or six days. We identified sets of common and stimuli-specific genes, the expression of which changed following stimulation with LPS, TNF-alpha or CD40L. A dynamic analysis of the entire DC differentiation and maturation process showed that some important inflammatory and constitutive chemokines are transcribed in both immature and mature DC. The correlative expression kinetics of the gene pairs IL1R1/IL1R2, IL15/IL15RA, DC-SIGN/ICAM-2 and DC-SIGN/ICAM-3 imply that they all play crucial roles in mediating DC functions. Thus, our analysis with focused microarrays shed light on the transcriptional kinetics of DC differentiation and maturation, and this method may also prove useful for identifying novel marker genes involved in DC functions. PMID- 19554265 TI - Web application for studying the free energy of binding and protonation states of protein-ligand complexes based on HINT. AB - A public web server performing computational titration at the active site in a protein-ligand complex has been implemented. This calculation is based on the Hydropathic interaction noncovalent force field. From 3D coordinate data for the protein, ligand and bridging waters (if available), the server predicts the best combination of protonation states for each ionizable residue and/or ligand functional group as well as the Gibbs free energy of binding for the ionization optimized protein-ligand complex. The 3D structure for the modified molecules is available as output. In addition, a graph depicting how this energy changes with acidity, i.e., as a function of added protons, can be obtained. This data may prove to be of use in preparing models for virtual screening and molecular docking. A few illustrative examples are presented. In beta secretase (2va7) computational titration flipped the amide groups of Gln12 and Asn37 and protonated a ligand amine yielding an improvement of 6.37 kcal mol(-1) in the protein-ligand binding score. Protonation of Glu139 in mutant HIV-1 reverse transcriptase (2opq) allows a water bridge between the protein and inhibitor that increases the protein-ligand interaction score by 0.16 kcal mol(-1). In human sialidase NEU2 complexed with an isobutyl ether mimetic inhibitor (2f11) computational titration suggested that protonating Glu218, deprotonating Arg237, flipping the amide bond on Tyr334, and optimizing the positions of several other polar protons would increase the protein-ligand interaction score by 0.71 kcal mol(-1). PMID- 19554267 TI - The influence of JNK and P38 MAPK inhibition on IL-12P40 and IL-23 production depending on IL12B promoter polymorphism. AB - The interleukin-12p40 gene (IL12B) encodes the p40 polypeptide chain, which, together with p19, composes IL-23. A bi-allelic promoter polymorphism (IL12Bpro) located at -2703 bp of the transcription initiation site has been reported to show associations with IL-12p40 production. To elucidate the dependence of IL 12p40 and IL-23 production on IL12Bpro polymorphism in relation to MAPK signal transduction pathways, we examined the effect of JNK and p38 inhibition on the secretion of these cytokines by stimulated peripheral blood mononuclear cells (PBMC) from healthy donors with 1.1 and 1.2/2.2 IL12Bpro genotypes. Stimulation with LPS and C3bgp resulted in approximately equal IL-12p40 production from PBMC with the 1.1 and 1.2/2.2 genotypes. The inhibition of JNK and p38 before stimulation significantly upregulated IL-12p40 production by PBMC with the 1.1 genotype, but did not influence IL-12p40 production from PBMC with the 1.2/2.2 genotype. Cultures of PBMC with the 1.1 genotype produced significantly more IL 12p40 than PBMC with the 1.2/2.2 genotype after stimulation with PHA. Inhibition of p38 kinase upregulated p40 production only in cultures with the 1.1 genotype. Decreased IL-23 production was observed in C3bgp-stimulated cultures after the inhibition of p38 regardless of the genotype of the tested cells. We concluded that IL-12p40 and IL-23 expression, which is mediated by the p38 and JNK intracellular signaling pathways, is influenced by IL12Bpro polymorphism. PMID- 19554268 TI - Mechanisms for the formation of membranous nanostructures in cell-to-cell communication. AB - Cells interact by exchanging material and information. Two methods of cell-to cell communication are by means of microvesicles and by means of nanotubes. Both microvesicles and nanotubes derive from the cell membrane and are able to transport the contents of the inner solution. In this review, we describe two physical mechanisms involved in the formation of microvesicles and nanotubes: curvature-mediated lateral redistribution of membrane components with the formation of membrane nanodomains; and plasmamediated attractive forces between membranes. These mechanisms are clinically relevant since they can be affected by drugs. In particular, the underlying mechanism of heparin's role as an anticoagulant and tumor suppressor is the suppression of microvesicluation due to plasma-mediated attractive interaction between membranes. PMID- 19554269 TI - [Perioperative myocardial damage in non-cardiac surgery patients]. AB - Perioperative myocardial damage occurs with a high incidence depending on the operative procedure and the patients examined and is considered to be among the most relevant risk factors for increased perioperative morbidity and mortality in patients undergoing non-cardiac surgery. The pathophysiology of myocardial damage in the perioperative period is still not well understood. Both ischemia with and without acute coronary occlusion and non-ischemic stimuli can put a substantial strain on the heart in the perioperative period. However, in many cases the clinical presentation does not allow a clear differentiation between ischemic and non-ischemic myocardial damage. In the majority of cases perioperative myocardial infarctions occur with only mild or even without any clinical symptoms. This is probably due to a considerable difference in phenotype and pathophysiology between perioperative and non-perioperative myocardial infarctions. As a result of this unexplained etiology of perioperative myocardial infarction it remains an open question whether the contemporary diagnostic and therapeutic recommendations for the acute coronary syndrome can be extrapolated to the perioperative situation. The present review reflects the current state of knowledge and presents an optional approach to the diagnosis and therapy of perioperative myocardial injury. PMID- 19554270 TI - [Target-controlled infusion. Clinical relevance and special features when using pharmacokinetic models]. AB - Since its commercial introduction in 1996, target-controlled infusion (TCI) has become an established technique for administration of intravenous anaesthetics. Modern TCI systems, however, are characterized by an increasing number of additional options and features, such as the choice between different pharmacokinetic models and modes of application, which may confuse the less experienced user. This review describes the differences between pharmacokinetic models, modes of application and the effect of covariates as well as the consequences for dosing. The aim is to explicate for the user of modern TCI systems the underlying scientific concepts and the relevance for clinical practice. PMID- 19554272 TI - [Surgical technique in cochlear implantation]. AB - The cochlear implant (CI) has become a standard option for treating prelingually deaf children. But postlingual late deafness in adults is becoming increasingly common. In addition, hybrid implantation with a CI and a hearing aid in the same ear has come into focus, which demands a soft insertion technique that spares the apical parts of the cochlea. Also, the chorda tympani should be saved, especially in bilateral implantations, which are gaining importance because improved speech discrimination in noisy conditions is seen as proven today. Control of the electrode position intraoperatively with intraoperative computed tomography can further increase the safety and reliability of the position. The position and length of the skin incision is a more aesthetic issue. Future developments will include fully implantable CIs and navigation-assisted, minimally invasive drilling of a hole from the surface of the skull into the cochlea. Bioactive, neurotrophic-drug-releasing electrode designs for improved and sustainable connectivity to the neurons may become applicable. PMID- 19554271 TI - [Cricoid pressure]. AB - The vast majority of anaesthetists considers application of cricoid pressure for reasons of patient safety an integral part of rapid sequence induction. Cricoid pressure is applied with the idea that it will prevent regurgitation of gastric content into the pharynx, thereby reducing the incidence of pulmonary aspiration. This review describes the background of the introduction of cricoid pressure into clinical practice, analyzes published data concerning clinical relevance of perioperative pulmonary aspiration and efficacy of cricoid pressure in reducing it, discusses problems associated with its use, assesses knowledge and technical performance of cricoid pressure and presents various recent recommendations regarding application of cricoid pressure. The combination of complete lack of evidence for the efficacy of cricoid pressure in preventing pulmonary aspiration and numerous reports of clinically relevant interference with airway management during its use, seriously question the rationale of recommending the general use of cricoid pressure during rapid sequence induction. PMID- 19554273 TI - [Sequential information processing in children with and without auditory processing disorder]. AB - BACKGROUND: It is a controversial issue whether sequential processing in children with auditory processing disorders (APD) is a unimodal auditory impairment. PATIENTS AND METHODS: Normal achieving controls (n=12; mean age: 101.1 months; SD 20.3) and children referred to clinical facilities for assessment including children with monosymptomatic APD (n=25; mean age: 90.8 months; SD 9.8), children with developmental language disorder (DLD]) + APD (n=11; mean age: 89.5 months; SD 14.9), children with dyslexia + APD (n=10; mean age: 113.8 months; SD 17.1) were compared using the subtest digit recall of the German version of the K-ABC (Melchers & Preuss 2001) and the K-ABC subtest hand movements in the visual modality. RESULTS: On average all groups tended to perform alike in visual sequential processing and memory (T-score>50). Normal controls (non-APD) showed on average the best test performance (T-score 57.4; SD 10.5), but the mean performance did not differ significantly from children with APD. All groups also presented on average normal test performance in the auditory modality however, children of the clinical groups were below the age mean (DLD + APD: T-W 43.1; SD 6.8; monosymptomatic APD: T-W 45.8; SD 9.8; dyslexia + APD: 46.7; SD5.8). Children with DLD + APD and those with monosymptomatic APD exhibited a significantly lower but not poor performance in auditory processing of sequences compared to their performance in visual sequential processing. The results of auditory and visual processing of sequences were significantly correlated in the clinical groups DLD + APD (0.58) and monosymptomatic APD (0.34), but not in the smallest group dyslexia + APD (0.48) and in normal achieving controls (0.32). CONCLUSION: On the basis of the results of the present study it may be concluded that a normal test performance on visual measure does not exclude a bimodal or pansensory seriation disorder in children with APD. It does, however, indicate the existence of a primary auditory sequential processing deficit, because at least the processing of phonological and visual stimuli was not separately carried out in children with DLD + APD. PMID- 19554275 TI - Frequency of hospitalization for angina pectoris, stroke, and peripheral venous thrombosis and its relationship to elements in rainwater in Opole Voivodship, Poland, during 2000-2002. AB - The content of elements in rainwater is an indirect indicator of its occurrence in air dust. This is sometimes referred to as rain fallout and is investigated in applied environmental pollution monitoring schemes. The annual content of elements in rainwater may be recognized as good index for assessing influence of those environmental factors on human body. The possible relationship between the concentrations of selected elements in rainwater and the frequency of hospitalization by reason of angina pectoris, stroke, and peripheral venous thrombosis was investigated in the Opole Voivodship (Poland) area during the period 2000-2002. There is a relatively high or partly significant correlation between frequency of hospitalization by reason of these conditions and content of lead, cadmium, chromium, zinc, and chloride in rainwater. Significant gender dependent differences were observed only in peripheral venous thrombosis, where important correlations with lead, cadmium, and chromium were found only in men. PMID- 19554274 TI - Efficacy of connective tissue massage and Mc Mennell joint manipulation in the rehabilitative treatment of the hands in systemic sclerosis. AB - Rehabilitation may contribute to the management of systemic sclerosis (SSc) dealing with disabilities due to hand involvement. The aim of this study is to evaluate the efficacy of a rehabilitation programme based on the combination of connective tissue massage and Mc Mennell joint manipulation specifically conceived for SSc patients' hands. Forty SSc patients were enrolled: 20 (interventional group) were treated for a 9-week period (twice a week, 1 h per session) with a combination of connective tissue massage, Mc Mennell joint manipulation and home exercise programme, and 20 (control group) were assigned only to home exercise programme. Patients of both groups were assessed at baseline (T0), after 9 week (T1) and at a 9 weeks follow-up (T2). They were evaluated for quality of life by SF-36 and Health Assessment Questionnaire (HAQ), hands involvement by Hand Mobility in Scleroderma (HAMIS) test, Cochin hand functional disability scale and the measurements of ROM. In the interventional group, fist closure, HAMIS test and Cochin hand functional disability scale improved at the end of the treatment (p < 0.0001) as well as HAQ, Physical Synthetic Index (PSI) and Mental Synthetic Index (MSI) of SF-36 scores (HAQ and PSI, p < 0.0001; MSI, p < 0.001). In the control group, the programme of home daily exercises improved only fist closure at the end of the treatment (p < 0.0001). The combination of connective tissue massage, Mc Mennell joint manipulation and home exercise programme is effective in the rehabilitative treatment of SSc hands. This combined treatment may lead to an improvement of hand function and quality of life. PMID- 19554279 TI - Abstracts of the International Conference on Ageing. October 29-31, 2008. Beijing, China. PMID- 19554276 TI - Effects of chromium(III) picolinate on cortisol and DHEAs secretion in H295R human adrenocortical cells. AB - Dietary chromium(III) picolinate (CrPic) effects on circulating steroid hormones have been reported in various experimental animals. However, direct effects of CrPic on adrenocortical steroidogenesis are uncertain. Therefore, the objective was to determine the effects of CrPic on cortisol and dehydroepiandrosterone sulfate (DHEAs) secretion from H295R cells. In experiment 1, a 24-h exposure to CrPic (0 to 200 microM) had both linear (p < 0.001) and quadratic (p < 0.001) effects on cortisol secretion from forskolin-stimulated cells with the highest cortisol secretion at 0.1 microM of CrPic and the lowest at 200 microM of CrPic. In experiment 2, a 48-h exposure to CrPic (200 microM) decreased cortisol (p < 0.07) release from forskolin-stimulated cells during a 24-h collection period. In experiment 3, a 48-h exposure to CrPic (100 microM) decreased cortisol (p < 0.05) and DHEAs (p < 0.01) from forskolin-stimulated cells during a 24-h sampling period. In experiment 4, a 24-h exposure to forskolin followed by a 24-h exposure to both forskolin and CrPic (100 and 200 microM) decreased both cortisol and DHEAs secretion (p < 0.01). This study suggests that at high concentrations, CrPic inhibits aspects of steroidogenesis in agonist-stimulated adrenocortical cells. PMID- 19554280 TI - Protein synthesis in nerve terminals and the glia-neuron unit. AB - The progressive philogenetic lengthening of axonal processes and the increase in complexity of terminal axonal arborizations markedly augmented the demands of the neuronal cytoplasmic mass on somatic gene expression. It is proposed that in an adaptive response to this challenge, novel gene expression functions developed in the axon compartment, consisting of axonal and presynaptic translation systems that rely on the delivery of transcripts synthesized in adjacent glial cells. Such intercellular mode of gene expression would allow more rapid plastic changes to occur in spatially restricted neuronal domains, down to the size of individual synapses. The cell body contribution to local gene expression in well differentiated neurons remains to be defined. The history of this concept and the experimental evidence supporting its validity are critically discussed in this article. The merit of this perspective lies with the recognition that plasticity events represent a major occurrence in the brain, and that they largely occur at synaptic sites, including presynaptic endings. PMID- 19554281 TI - Organizational dynamics, functions, and pathobiological dysfunctions of neurofilaments. AB - Neurofilament phosphorylation has long been considered to regulate their axonal transport rate, and in doing so it provides stability to mature axons. We evaluate the collective evidence to date regarding how neurofilament C-terminal phosphorylation may regulate axonal transport. We present a few suggestions for further experimentation in this area, and expand upon previous models for axonal NF dynamics. We present evidence that the NFs that display extended residence along axons are critically dependent upon the surrounding microtubules, and that simultaneous interaction with multiple microtubule motors provides the architectural force that regulates their distribution. Finally, we address how C terminal phosphorylation is regionally and temporally regulated by a balance of kinase and phosphatase activities, and how misregulation of this balance might contribute to motor neuron disease. PMID- 19554282 TI - Myosin motor proteins in the cell biology of axons and other neuronal compartments. AB - Most neurons of both the central and peripheral nervous systems express multiple members of the myosin superfamily that include nonmuscle myosin II, and a number of classes of unconventional myosins. Several classes of unconventional myosins found in neurons have been shown to play important roles in transport processes. A general picture of the myosin-dependent transport processes in neurons is beginning to emerge, although much more work still needs to be done to fully define these roles and establish the importance of myosin for axonal transport. Myosins appear to contribute to three types of transport processes in neurons; recycling of receptors or other membrane components, dynamic tethering of vesicular components, and transport or tethering of protein translational machinery including mRNA. Defects in one or more of these functions have potential to contribute to disease processes. PMID- 19554283 TI - Antibody phage display: overview of a powerful technology that has quickly translated to the clinic. AB - Antibody-based immunologic reagents are useful for identifying, isolating, or eliminating cells with particular characteristics related to different diseases. Phage display is a highly valuable technique for antibody selection related to this purpose. In brief, a diverse group of antibody genes prepared from a patient or generated in vitro are inserted into a phagemid vector or the phage genome so that when the protein is expressed, it becomes anchored on the surface of the phage by fusion to a coat protein. A diverse library of recombinant antibodies is generated in this way and can then be exposed or panned on the antigen of interest, typically, this being a molecule associated with a particular pathological condition. Phage that carry proteins or peptides bind preferentially to the target and can thus be isolated from the library. The viruses that are recovered in this way also carry the gene for the binding moiety facilitating its over-expression or manipulation. Recent reviews highlight key milestones in the development of antibody libraries and their screening by phage display, and the impact of these technologies on drug discovery seems assured. PMID- 19554284 TI - Design and construction of synthetic phage-displayed Fab libraries. AB - Diversity-the variability carried by the amino acid sequences of a synthetic antibody library-can be generated by synthetic degenerate oligonucleotides. One can experiment with different diversity designs in the variable domains of light and heavy chains (V(H) and V(L)) to generate antibody libraries with different properties. The ability to precisely define the final diversity of a library facilitates the process of isolating, characterizing, and optimizing an antibody lead. Here we describe detailed protocols for the design and construction of phage-displayed synthetic antibody libraries in which diversity is generated in the complementarity determining regions (CDRs) of the V(H) of a single humanized bivalent Fab scaffold. The example used in the protocol provides a general methodology for generation of libraries with engineered CDR diversity that can be applied to a template antibody sequence of choice. PMID- 19554285 TI - Affinity isolation of antigen-specific circulating B cells for generation of phage display-derived human monoclonal antibodies. AB - A method is described for affinity isolation of antigen-specific circulating B cells of interest for subsequent generation of immune antibody phage display libraries. This approach should overcome the problem of low yields of monoclonal antibodies of interest in the libraries generated from peripheral blood lymphocytes caused by the low abundance of antigen-specific B cells in the circulation. The preselection of B cells is based on the specificity of the surface Ig receptor and is accomplished using the antigen of interest conjugated to magnetic beads. This method should significantly increase the frequency of antibody phage particles of interest in the library and allow for efficient isolation monoclonal antibodies with the predefined specificity. PMID- 19554286 TI - Construction of phage antibody repertoires from the blood of West Nile virus infected donors. AB - A method for the construction of West Nile virus immune donor antibody repertoires is described. B cells are harvested from a suitable donor and the antibody variable genes are amplified using polymerase chain reaction (PCR). The PCR fragments are cloned in a phage display vector to construct a repertoire that can be used in panning procedures to identify many unique monoclonal antibodies. PMID- 19554287 TI - Anti-beta(2)GP-I and anti-prothrombin antibodies generated by phage display. AB - This chapter describes the construction and screening of a library of single chain variable fragments (svFv) derived from patients with autoimmune disease. The methods cover the isolation of mononuclear cells from peripheral blood, preparation of RNA, and recovery of immunoglobulin-coding sequences by polymerase chain reaction (PCR). Cloning into a phage display vector and screening of the scFv display library by a simple panning procedure are described. These methods are applicable to library construction from any patient group or (with alternative primer sets) any mammalian species. PMID- 19554288 TI - The generation and selection of single-domain, v region libraries from nurse sharks. AB - The cartilaginous fish (sharks, skates, and rays) are the oldest phylogenetic group in which a human-type adaptive immune system and immunoglobulins (Igs) have been found. In addition to their conventional (heavy-light chain heterodimeric) isotypes, IgM and IgW, sharks produce the novel isotype, IgNAR, a heavy chain homodimer that does not associate with light chains. Instead, its variable (V) regions act as independent, soluble units in order to bind antigen. In this chapter, we detail our immunization protocol in order to raise a humoral IgNAR response in the nurse shark (Ginglymostoma cirratum) and the subsequent cloning of the single-domain V regions from this isotype in order to select antigen specific binders by phage display. PMID- 19554289 TI - The isolation of scFvs against small target molecules. AB - Phage display has the capacity to rapidly isolate recombinant antibodies against protein targets and other molecules of significant size. However, there is no obvious lower limit to the power of the selection methods: this chapter describes how the techniques of phage display can be adapted to allow the isolation of antibodies against very small compounds. Antibodies generated in this way have many uses including the detection and quantitative analysis of the target chemical moiety in samples such as foods, water, and body fluids. PMID- 19554290 TI - Display and selection of scFv antibodies on HEK-293T cells. AB - We describe a human cell display strategy to isolate high-affinity single-chain antibody fragments (scFvs) specific for CD22 for the treatment of B-cell malignancies. Our strategy uses flow cytometry and human embryonic kidney 293T (HEK-293T) cells that are widely used for transient protein expression. Flow cytometry enhances the screen's sensitivity thereby allowing us to isolate high affinity scFvs. Using human cell display, one could isolate and engineer scFvs, single domains, Fabs, or whole IgGs for increased affinity and other biological functions. PMID- 19554291 TI - Isolation of scFvs that inhibit the NS3 protease of hepatitis C virus by a combination of phage display and a bacterial genetic screen. AB - The need for inhibitors for enzymes linked with microbial infection, specifically the NS3 protease of hepatitis C virus (HCV), inspired us to develop a unique, rapid and easy color-based method described herein. The NS3 serine protease of HCV has a role in processing viral polyprotein and it has been implicated in interactions with various cell constituents, resulting in phenotypic changes including malignant transformation. NS3 is currently regarded a prime target for antiviral drugs.We established a genetic screen that is based on coexpression of NS3, a beta-galactosidase reporter that is cleavable by NS3, and potential inhibitors within the same bacterial cell. A single-chain antibody (scFv) library was prepared from spleens of NS3-immunized mice and the screen was used to isolate a panel of protease-inhibiting scFvs. Candidate scFvs were validated for inhibitory activity using an o-nitrophenyl-beta-galactoside (ONPG) hydrolysis assay.The methods can be used more generally to isolate protease-inhibiting cytoplasmic intrabodies able to inhibit proteases or other activities that can be linked with the phenotype of Escherichia coli. PMID- 19554292 TI - Guided selection methods through chain shuffling. AB - We provide procedures for the panning of fully humanized Fab antibodies using guided selection. Human heavy and light chain genes are amplified. A parental light chain is cloned into a phage display vector and combined with the heavy chain library. After several rounds of panning, positive clones are identified and the heavy chain sequences that are recovered are combined with light chains for further selection by phage display. Human Fab antibodies are obtained that bind the same epitope as the parental antibody. PMID- 19554293 TI - Sequential antigen panning for selection of broadly cross-reactive HIV-1 neutralizing human monoclonal antibodies. AB - Many phage display techniques drive selection toward the isolation of highly specific antibodies. However, the identification of monoclonal antibodies that are cross-reactive has implications for the development of diagnostics, therapeutics, and vaccines against pathogens or cancer cells that are able to rapidly generate variants and escape mutants. To identify human monoclonal antibodies with high activity against HIV and broad-spectrum activity, we developed a technique termed sequential antigen panning. This methodology could be used to isolated recombinant antibodies against any antigen that shares epitopes with other antigens. PMID- 19554294 TI - Selection of antibodies able to rapidly enter mammalian cells. AB - This chapter outlines a protocol for the selection by phage display of single chain variable antibody fragments with dual properties-specificity for tumor cells and the ability to be internalized. The protocol is based on a direct incubation of living target cells with antibody phage display libraries under conditions that allow active endocytosis of phage particles by cancer cells as well as recovery of intracellular phage particles that retain infectivity. This "functional" selection helps avoid the isolation of irrelevant phages that may be obtained when selection is performed on heterogeneous material as a source of antigen. Internalizing antibodies recovered from human antibody repertoires are promising reagents for various therapeutic applications including the delivery of drugs to the cytosol of targeted cell populations. PMID- 19554295 TI - Identification of the specificity of isolated phage display single-chain antibodies using yeast two-hybrid screens. AB - A method is described for the identification of the antigen recognised by an scFv isolated from an antibody phage display library using selection against a complex mixture of proteins (e.g. intact cells, purified cell surface membranes, and tissue sections). The method takes advantage of a yeast two-hybrid system that additionally allows for reorganization of post-translational modifications to the bait and target proteins. This technique is therefore especially useful for identifying surface-expressed antigens. PMID- 19554296 TI - Generation of bispecific and tandem diabodies. AB - Conventionally, antibody phage display has been used to isolate recombinant antibodies that are monovalent in their interaction with target antigens. These antibodies can be reengineered for expression in mammalian cell culture as full length, monospecific immunoglobulins. An emerging branch of research has sought to generate bivalent recombinant antibodies by manipulating the length of the linker separating heavy- and light-chain variable domains in single-chain Fv proteins, thereby promoting inter-scFv interaction and the formation of "diabodies." With careful control, this can generate scFv-based proteins able to bind two very distinct targets, "bispecific diabodies." Further manipulation enables the assembly of higher-order complexes. PMID- 19554297 TI - Intrabody expression in eukaryotic cells. AB - We describe procedures for intracellular expression of scFv in eukaryotic cells. Starting from a scFv gene cloned in a phage-display vector, we describe the cloning step into a mammalian expression vector, the transient transfection of a HeLa cell line, and the monitoring of intrabody expression by immunofluorescence staining and FACS analysis. PMID- 19554298 TI - High-level periplasmic expression and purification of scFvs. AB - The isolation of recombinant antibodies by phage display naturally leads to experiments to evaluate their biological and immunological properties. Although crude preparations may have their value in initial studies, the need often exists for highly purified protein that can be tested in vivo. This chapter describes methods to generate high yields of scFv from bacterial cultures and to purify protein to the degree of homogeneity required for the most exacting analysis. PMID- 19554299 TI - Expression of single-chain Fv fragments in E. coli cytoplasm. AB - The most frequently used approach to produce single-chain Fv fragments (scFv) and Fab in Escherichia coli is to express them in the periplasm of the bacteria. We present here an alternative procedure that uses cytoplasmic expression of soluble active scFv. This can be accomplished by using either specially engineered E. coli strains or hyperstable scFvs. PMID- 19554300 TI - High-level expression of a phage display-derived scFv in Pichia pastoris. AB - Numerous techniques are available for investigating protein-ligand interactions. The phage display technique is one such method routinely used to identify antibody-antigen interactions and has the benefit of being easily adaptable to high-throughput screening platforms. Once identified, antigen-binding domains on fragment antibodies or single-chain fragment antibodies (scFv) can be expressed and purified for further studies. In this chapter, we describe a method for high level expression of a phage display-derived scFv in Pichia pastoris. The phage display-derived antibody A33scFv recognizes a cell surface glycoprotein (designated A33) expressed in colon cancer that serves as a target antigen for radioimmunoimaging and/or immunotherapy of human colon cancer. The expression and purification of A33scFv was optimized for the methylotrophic yeast P. pastoris. P. pastoris with a Mut(S) phenotype was selected to express A33scFv under regulation of the methanol-inducible AOX1 promoter. Here we describe a large scale fed-batch fermentation process with an efficient online closed-loop methanol control for the production of the recombinant protein. Purification of A33scFv from clarified culture medium was done using a two-step chromatographic procedure using anion exchange and hydrophobic interaction chromatography, resulting in a final product with more than 90% purity. This chapter provides protocols that can be used as a base for process development of recombinant protein expression in P. pastoris and purification of these proteins for use in further functionality studies and in diagnostic and therapeutic applications. PMID- 19554301 TI - DNA deaminases: AIDing hormones in immunity and cancer. AB - It is well established that hormones can cause cancer, much less known is how they induce this change in our somatic cells. This review highlights the recent finding that estrogen can exert its DNA-damaging potential by directly activating DNA deaminases. This recently discovered class of proteins deaminate cytosine to uracil in DNA, and are essential enzymes in the immune system. The enhanced production of a given DNA deaminase, induced by estrogen, can lead not only to a more active immune response, but also to an increase in mutations and oncogenic translocations. Identifying the direct molecular link between estrogen and a mutation event provides us with new targets for studying and possibly inhibiting the pathological side-effects of estrogen. PMID- 19554303 TI - [The bought patient : The ethical value of per capita flat rates from a legal viewpoint]. AB - From the decisions of the German Medical Council for safeguarding medical independence and the new version of section 18 paragraph 1 MBO with reference to the cooperation between hospitals and physicians for patient relationship management, it can be derived that clear competitive and professional limits exist in the competition for patients within the cooperation of hospitals and physicians. The ethical and professional foundation serving the patient that physicians must make decisions regarding the patient in an independent way free from commercial influences, plays a key role for the legal assessment of models for patient allocation irrespective of the planned increased commercial and competitive nature of the service provided by social health insurance caused by the health reforms. PMID- 19554302 TI - OSBPL10, a novel candidate gene for high triglyceride trait in dyslipidemic Finnish subjects, regulates cellular lipid metabolism. AB - Analysis of variants in three genes encoding oxysterol-binding protein (OSBP) homologues (OSBPL2, OSBPL9, OSBPL10) in Finnish families with familial low high density lipoprotein (HDL) levels (N = 426) or familial combined hyperlipidemia (N = 684) revealed suggestive linkage of OSBPL10 single-nucleotide polymorphisms (SNPs) with extreme end high triglyceride (TG; >90th percentile) trait. Prompted by this initial finding, we carried out association analysis in a metabolic syndrome subcohort (Genmets) of Health2000 examination survey (N = 2,138), revealing association of multiple OSBPL10 SNPs with high serum TG levels (>95th percentile). To investigate whether OSBPL10 could be the gene underlying the observed linkage and association, we carried out functional experiments in the human hepatoma cell line Huh7. Silencing of OSBPL10 increased the incorporation of [(3)H]acetate into cholesterol and both [(3)H]acetate and [(3)H]oleate into triglycerides and enhanced the accumulation of secreted apolipoprotein B100 in growth medium, suggesting that the encoded protein ORP10 suppresses hepatic lipogenesis and very-low-density lipoprotein production. ORP10 was shown to associate dynamically with microtubules, consistent with its involvement in intracellular transport or organelle positioning. The data introduces OSBPL10 as a gene whose variation may contribute to high triglyceride levels in dyslipidemic Finnish subjects and provides evidence for ORP10 as a regulator of cellular lipid metabolism. PMID- 19554304 TI - [Effects of the new further training regulations on the training behavior of assistant doctors]. AB - During the 106th "Deutschen Arztetag" (German Medical Assembly) in 2003 new model further training regulations were finalized. Further training was to be more clearly arranged and the targets more clearly defined. There are no uniform training regulations for Germany. The new training regulations promise more clarity: the numbers of the individual interventions and investigations are no longer specified. Instead operation groups which are connected with respect to content and organization in a defined minimum are to be identified. The new further training regulations are geared to a process orientation. For the first time trainers and trainees have the possibility to actively work together in the further training process. The joint logbook"Urological further training" from the GeSRU (German Society of Residents in Urology), DGU (German Society for Urology) and BDU (Professional Association of German Urologists) is a milestone in Germany. The further training will be continuously improved through constant evaluation. PMID- 19554305 TI - [Local treatment of prostate cancer using thermal-ablative energy]. AB - Thermal and thermal-ablative procedures for treating prostate cancer have been investigated systematically since approximately 1980 (apart from some historical predecessors), and numerous experimental and clinical reports have been published on this subject. Various technologies have been used, including transurethral ablation of prostatic tissue using laser or microwave energy, interstitial application of laser or microwave energy, and inductive heating of previously implanted thermoseeds or injected magnetic nanoparticles in a magnetic field. For all of these procedures, clinical studies with a total of some 350 patients have been performed. However, the results cannot be judged correctly because of a lack of adequate control parameters for the older studies and inadequately short follow-up of all studies. Conclusions regarding treatment-related morbidity seem to be possible, with a generally positive impression and low rates of adverse effects. But before such results can be generalized, patient selection bias and the technology standards that existed when the studies were performed must be taken into consideration. Various papers are reviewed and summarized. In the author's opinion, the different options for thermal and thermal-ablative treatment of prostate cancer are very promising, but in light of the existing standard procedures, feasibility must not overrule reasonableness. PMID- 19554306 TI - Relationships, trust, decision-making and quality of care in a paediatric intensive care unit. AB - OBJECTIVE: To assess the care-giving practices of health-care practitioners in the paediatric intensive care unit (PICU) through their qualitative insights, reflections and experience in participatory action research. DESIGN AND METHODS: Qualitative research in the form of 'participatory action research' was used to gather data from three sources within the unit: focus groups within disciplines, observations within the PICU, and semi-structured interviews. All staff members were active collaborators and equal stakeholders in the decision-making process, research and feedback. SETTING: The paediatric intensive care unit (PICU) of the Red Cross War Memorial Children's Hospital (RCWMCH). PARTICIPANTS: All staff members from various disciplines working in the PICU. RESULTS: Staff members described problems with respect to relationships, trust and decision-making within care-giving practices. CONCLUSION: The study qualitatively describes how poor communication amongst staff members in respect of relationships and decision making impacted on trust and how this tended to compromise care-giving practices in the PICU. The data suggested that this was more evident in informal rather than formal clinical decision-making procedures. The strength of the study was that the participatory action design in the research allowed staff members to address the very dynamics that they themselves cited as problematic. PMID- 19554308 TI - Arthroscopic suture anchor repair for lateral patellar instability. AB - Several all-arthroscopic techniques have been described to address patellar instability. Most arthroscopic procedures focus on soft tissue plication or "tightening" of the medial retinacular structures to correct lateral patellar instability. We found these techniques to be ineffective when the medial stabilizers were avulsed from the patella. As a result, we have developed an arthroscopic technique to repair medial patellofemoral ligament avulsions to the patella using suture anchors. PMID- 19554307 TI - Point-of-care assessment of microvascular blood flow in critically ill patients. AB - OBJECTIVE: Sublingual microvascular videomicroscopy can assess tissue perfusion in critically ill patients; however, data analysis is currently limited to delayed off-line evaluation. We hypothesized that a real-time point-of-care (POC) determination of the microcirculatory flow index (MFI), an established metric for assessing microcirculatory perfusion, agrees well with the conventional off-line analysis. DESIGN: Prospective observational study. SETTING: Urban academic intensive care unit. PARTICIPANTS: A heterogeneous population of critically ill patients. MEASUREMENTS AND RESULTS: A single operator performed side stream darkfield videomicroscopy of the sublingual microcirculation and made a POC determination of MFI in real-time on a portable bedside monitor by assigning a score (0 = no flow to 3 = normal) to each quadrant of the image and averaging the four values. After image processing, de-identification and randomization, the same operator, blinded to the previous interpretation, repeated the MFI assessment by viewing an AVI-formatted image sequence on a 94 cm 1,080 pixel LCD monitor (reference standard). There were 205 paired measurements in 18 subjects. The POC and reference standard MFI had good agreement by Bland-Altman analysis [mean difference of -0.031, SD = 0.198 (95% CI, -0.43 to 0.37)]. The POC assessment was 94% sensitive and 92% specific for detecting impaired microvascular flow (defined a priori as an MFI < 2.5 based on previously published data). CONCLUSIONS: A POC determination of MFI had good agreement with conventional off-line analysis, and was highly sensitive and specific for detecting impaired microvascular flow. This real-time technique may be useful in future clinical trials targeting impaired microcirculatory perfusion in critically ill patients. PMID- 19554309 TI - The relationship between tibial slope and meniscal insertion. AB - Despite increasing interest in the anatomic importance of the meniscal insertion, little information is available regarding the relationship between the tibial slope and the meniscal insertion. Lateral radiographs and MRI sagittal images from 100 healthy and young patients were used in this study. Patients without deformation, meniscal pathology, or previous surgery to the ipsilateral knee were included in this study. We measured the angle between a line tangent to the medial and lateral tibial slope and the proximal tibial anatomical axis using a lateral radiographs. We also measured the angle between the tangent line to the medial and lateral tibial insertion of the meniscus and the proximal tibial anatomical axis using sagittal MRI images. The measurements were carried out twice by two observers. Inter-observer reliability ranged from 0.98 to 0.99 and intra-observer reliability ranged from 0.83 to 0.94. For each observer, the mean differences between measurements made using radiographs and MRI images were 16.4 degrees and 16.4 degrees on the lateral side, respectively, and 6.0 degrees and 5.9 degrees on the medial side, respectively. There was a statistically significant difference between measurements made using radiographs and MRI images (p < 0.001). However, the Pearson's correlation coefficient between the measurements made using radiographs and MRI images did not show a linear correlation. The measurements of posterior slope on lateral radiographs images and meniscal insertion on sagittal MRI images were reproducible and reliable. Differences in measurements ranged from 15 degrees to 17 degrees on the lateral side and from 5 degrees to 6 degrees on the medial side, with 95% confidence intervals. However, there was no statistical correlation between the measurements made using lateral radiographs and MRI images. PMID- 19554310 TI - Radial tears in the roots of the posterior horns of both the medial and lateral menisci combined with anterior cruciate ligament tear: a case report. AB - We report a rare case of radial tears in the roots of the posterior horns of both the medial and lateral menisci associated with a chronic anterior cruciate ligament tear. Treatment included an arthroscopic pullout suture combined with anterior cruciate ligament reconstruction. At the 3-year postoperative follow-up, a second-look arthroscopic examination showed the posterior horns of both menisci to be well healed on the tibia. Manual knee laxity tests were negative and no side-to-side difference was detected by a KT-1000 arthrometer. The patient was able to perform outdoor activities without residual symptoms. PMID- 19554311 TI - Arthroscopic treatment of acute acromioclavicular joint dislocation with double flip button. AB - The ideal treatment for acute acromioclavicular joint dislocation is still controversial, both in terms of indications and surgical technique. The clinical and radiographic outcomes of 16 patients affected by acute AC joint dislocation (type III-V) and arthroscopically treated with a coracoclavicular double flip button are presented. Despite the excellent clinical results both in terms of Constant score (mean 97 points) and patient satisfaction, at a mean follow-up of 31 months the radiographs showed partial loss of reduction due to distal migration of the flip button within the upper third of the clavicle in one-fourth of the cases. The technique presented here proved to be safe and minimally invasive while delivering good aesthetic results and allowing for the treatment of associated lesions. Furthermore, the technique could benefit from more advanced retention devices, which ought to reduce or avoid migration of the flip buttons. PMID- 19554312 TI - The role of preoperative MRI in knee arthroscopy: a retrospective analysis of 2,000 patients. AB - The aim of this study was to investigate, to what extent routine preoperative MRI scans could set the indications for knee arthroscopies and reduce the number of diagnostic arthroscopies. For this retrospective cohort study, 1,000 patients who had knee arthroscopies documented in 1994/1995 were compared with 1,000 patients that were treated in 2004/2005. The preoperative diagnoses that gave indications for knee arthroscopy were compared with the intraoperative findings. The congruence of preoperative diagnosis with the intraoperative findings was evaluated comparing both study populations. The number of patients who were referred to orthopaedic trauma surgeons with MRI increased from 24% to 56%. A high congruence of preoperative diagnosis and intraoperative findings was found in 49% in 1994/1995 and 55% in 2004/2005. However, regarding the most important outcome parameter, the number of diagnostic arthroscopies, no improvement was found (3% in both periods). The presented data suggests that MRI scans are not routinely necessary as an indication for knee arthroscopy, as clinical examination and plain radiograph are sufficient. However, MRI scans do allow a more detailed characterization of the expected findings and can therefore be helpful in therapy planning. PMID- 19554313 TI - Quantitation and identity confirmation of residues of quinolones in tilapia fillets by LC-ESI-MS-MS QToF. AB - A method for simultaneous determination of flumequine (FLM), oxolinic acid (OXO), sarafloxacin (SAR), danofloxacin (DAN), enrofloxacin (ENR), and ciprofloxacin (CIP) in tilapia (Orechromis niloticus) fillets, using liquid chromatography tandem mass spectrometry (LC-ESI-MS-MS QToF) is presented. The quinolones were extracted from the food matrix with a solution of 10% trichloroacetic acid methanol (80:20 v/v) with ultrasonic assistance. Clean-up of the extract solution was performed by using polymeric solid-phase extraction cartridges. The LC separation was carried out on an octadecyl hybrid silica column (C18, 150 mm x 3 mm, 5 microm). The column temperature was set at 30 degrees C, and gradient elution (0.2 mL min(-1)) was performed using water and acetonitrile, both containing 0.1% of acetic acid, as mobile phase components. The analytes were ionized using electrospray in the positive polarity mode. The following analytical results were obtained: linearity was about 0.99 for all the quinolones; intra and inter-assay precision (RSD%) were lower than 12.7 and 20%, respectively; and recoveries were from 89 to 112%. The quantitation limits were below the maximum residue limits established for the analytes. The method is suitable for the determination of quinolone residues in fish fillets and the QToF technique made it possible to obtain m/z ratios with less than 10 ppm of error for each analyte. PMID- 19554314 TI - An SEC/MALS study of alternan degradation during size-exclusion chromatographic analysis. AB - Ultrahigh-molar-mass (M) polymers such as DNA, cellulose, and polyolefins are routinely analyzed using size-exclusion chromatography (SEC) to obtain molar mass averages, distributions, and architectural information. It has long been contended that high-M polymers can degrade during SEC analysis; if true, the inaccurate molar mass information obtained can adversely affect decisions regarding processing and end-use properties of the macromolecules. However, most evidence to the effect of degradation has been circumstantial and open to alternative interpretation. For example, the shift in SEC elution volume as a function of increased chromatographic flow rate, observed using only a concentration-sensitive detector, may be the result of degradation or of elution via a nondegradatory slalom chromatography mechanism. Here, using both concentration-sensitive and multiangle static light-scattering detection, we provide unambiguous evidence that the polysaccharide alternan actually degrades during SEC analysis. The decrease in molar mass and size of alternan with increasing flow rate, measured using light scattering, allows ruling out an SC mode of elution and can only be interpreted as due to degradation. These findings demonstrate the extreme fragility of ultrahigh-M polymers and the care that must be taken for accurate characterization. PMID- 19554315 TI - Patterning of 293T fibroblasts on a mica surface. AB - Controllable cell growth on the defined areas of surfaces is important for potential applications in biosensor fabrication and tissue engineering. In this study, controllable cell growth was achieved by culturing 293 T fibroblast cells on a mica surface which had been patterned with collagen strips by a microcontact printing (microCP) technique. The collagen area was designed to support cell adhesion and the native mica surface was designed to repel cell adhesion. Consequently, the resulting cell patterns should follow the micro-patterns of the collagen. X-ray photoelectron spectroscopy (XPS), water contact angle (WCA) measurement, atomic-force microscope (AFM) observation, and force-curve measurement were used to monitor property changes before and after the collagen adsorption process. Further data showed that the patterned cells were of good viability and able to perform a gene-transfection experiment in vitro. This technique should be of potential applications in the fields of biosensor fabrication and tissue engineering. PMID- 19554316 TI - Iron oxide/tantalum oxide core-shell magnetic nanoparticle-based microwave assisted extraction for phosphopeptide enrichment from complex samples for MALDI MS analysis. AB - A new type of metal-oxide-coated magnetic nanoparticles (NPs)--tantalum-oxide coated magnetic iron oxide (Fe3O4@Ta2O5) NPs--which are used as affinity probes for selectively trapping phosphopeptides from complex samples, is demonstrated in this study. In this approach, phosphopeptide enrichment was achieved by incubating the NPs with sample solutions under microwave heating within 1 min. The NP-target species conjugates were readily isolated from samples by magnetic separation followed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometric analysis. When using human serum as the sample, phosphorylated fibrinopeptide-A-derived ions are the only ions observed in the MALDI mass spectra after enrichment by the Fe3O4@Ta2O5 NPs. Furthermore, only phosphopeptides appear in the MALDI mass spectra after using the affinity probes to selectively trap target species from the tryptic digest of a cell lysate and milk sample. The results demonstrated that the Fe3O4@Ta2O5 NPs have the capability of selectively trapping phosphorylated peptides from complex samples. The detection limit of this approach for a phosphopeptide (FQpSEEQQQTEDELQDK) was approximately 10 fmol. PMID- 19554317 TI - Effects of carnosine on long-term plasticity of medial perforant pathway/dentate gyrus synapses in urethane-anesthetized rats: an in vivo model. AB - The objective of this study was to examine the effect of carnosine on the hippocampal-dependent learning in perforant pathway/dentate gyrus synapses. The experiments were carried out on adult rats. A bipolar stimulating electrode was placed to the medial perforant path and a double-barrel glass micropipette was placed in the dentate gyrus as the recording electrode. Artificial cerebrospinal fluid (to control group) or carnosine (0.1, 1 microg/microL) was infused into the dentate gyrus via one of the barrels of the glass electrode. Our results showed that the I/O curve of excitatory postsynaptic potential (EPSP) slope or population spike (PS) amplitude was not significantly shifted by carnosine. Although carnosine infused prior to high-frequency stimulation (HFS) decreased the slope of EPSP and amplitude of PS, when infused after HFS, no effect was observed. In the present study, we speculated that carnosine decreased LTP by inhibiting sGC activation. The present experiment provides the first evidence that carnosine may play a role in synaptic plasticity in dentate gyrus in vivo. PMID- 19554318 TI - Vision during head bobbing: are pigeons capable of shape discrimination during the thrust phase? AB - Many birds show a characteristic forward and backward head movement, while walking, running and sometimes during landing flight, called head bobbing. During the hold phase, the head of the bird remains stable in space, while during the thrust phase, the head is rapidly moved forward. Three main functions for head bobbing have been proposed: Head bobbing might have a biomechanical cause, it might serve depth perception via motion parallax, or it might be an optokinetic response that primarily serves image stabilization for improved vision during the hold phase. To investigate vision during the different phases and in particular to test for visual suppression during the saccadic thrust phase, we tested pigeons on a shape discrimination task, presenting the stimuli exclusively either in the hold phase, thrust phase or at random times. Results clearly demonstrate that shape discrimination is as good during the thrust phase as it is during the hold phase. PMID- 19554319 TI - Prehension synergies: a study of digit force adjustments to the continuously varied load force exerted on a partially constrained hand-held object. AB - We examined how the digit forces adjust when a load force acting on a hand-held object continuously varies. The subjects were required to hold the handle still while a linearly increasing and then decreasing force was applied to the handle. The handle was constrained, such that it could only move up and down, and rotate about a horizontal axis. In addition, the moment arm of the thumb tangential force was 1.5 times the moment arm of the virtual finger (VF, an imagined finger with the mechanical action equal to that of the four fingers) force. Unlike the situation when there are equal moment arms, the experimental setup forced the subjects to choose between (a) sharing equally the increase in load force between the thumb and VF but generating a moment of tangential force, which had to be compensated by negatively co-varying the moment due to normal forces, or (b) sharing unequally the load force increase between the thumb and VF but preventing generation of a moment of tangential forces. We found that different subjects tended to use one of these two strategies. These findings suggest that the selection by the CNS of prehension synergies at the VF-thumb level with respect to the moment of force is non-obligatory and reflects individual subject preferences. This unequal sharing of the load by the tangential forces, in contrast to the previously observed equal sharing, suggests that the invariant feature of prehension may be a correlated increase in tangential forces rather than an equal increase. PMID- 19554320 TI - Reexamining transaminase elevation in Phase I clinical trials: the importance of baseline and change from baseline. AB - PURPOSE: The aims of the present analysis were to determine prevalence of transaminase elevation in placebo-treated healthy volunteers in our historical phase I clinical trials and to assess which factors were associated with it. METHODS: We performed a retrospective analysis of alanine transaminase (ALT) levels in 481 placebo-treated healthy volunteers from 20 phase I trials by examining ALT elevation rates using the upper limit of normal values (ULN) as the cutoff as well as changes from baseline in actual ALT values. RESULTS: The ULN for ALT ranged from 32 to 72 IU/L across the studies. Although the overall ALT elevation rate (4.4%) from pooled datasets was low, the elevation rates were higher in more recent studies than in earlier ones. While elevation rates at baseline and during placebo treatment did not differ significantly, ALT maximal levels during placebo treatment were significantly higher than baseline levels. Moreover, baseline ALT levels were found to be more important in predicting ALT elevation during placebo treatment than demographic and study design factors. CONCLUSION: Baseline level and changes from baseline in transaminase are important variables to examine in addition to elevation above ULN for more reliably interpreting liver signals in Phase I clinical trials. PMID- 19554321 TI - Fetal MRI in the evaluation of fetuses referred for sonographically suspected neural tube defects (NTDs): impact on diagnosis and management decision. AB - INTRODUCTION: We hypothesized that magnetic resonance imaging (MRI) can assess fetuses with sonographically (ultrasonography (US))-suspected neural tube defects (NTD) that might influence their diagnoses and management decision. METHODS: Institutional review board approval and informed consents were obtained to perform MRI for 19 fetuses referred with US-suspected NTD. Prenatal imaging findings were correlated with management decision, postnatal clinical, postnatal imaging, and pathology. RESULTS: Prenatal MRI correctly ruled out US diagnosis of cephalocele in a fetus. In the other 18 fetuses, MRI detected detailed topography and contents of NTD sacs in five, added central nervous system (CNS) abnormalities that were not apparent on US in three, and confirmed non-CNS findings in three fetuses. MRI changed diagnosis of 3/19 fetuses (15.8%), caused minor change in diagnosis of 5/19 (26.3%), and did not influence US diagnosis of 11/19 fetuses (57.9%). MRI findings changed/modified management decision in 21% of the fetuses. CONCLUSION: Fetal MRI is an important adjunct to US in assessing NTD. It can identify topography and contents of sacs, add CNS and non-CNS findings, and influence management decision. PMID- 19554322 TI - Radiation exposure from pediatric head CT: a bi-institutional study. AB - BACKGROUND: Medical radiation from CT should be kept as low as reasonably achievable (ALARA), particularly in young patients. OBJECTIVE: To examine radiation dose from head CT in children in a trauma center (TC) and a regional children's hospital (RCH). MATERIALS AND METHODS: A random sample of 240 children (0-3, 4-9, 10-14 years of age) from the TC were compared with a similar cohort from the RCH. All children had undergone at least one head CT scan without contrast enhancement; data from PACS and Department of Radiology Information System were used to estimate normalized effective dose (ED). Lifetime attributable risk of cancer incidence was estimated using the Biologic Effects of Ionizing Radiation (BEIR) VII report. RESULTS: The mean normalized ED was significantly higher in the youngest children at the TC (2.74 mSv in those aged 0 3 years vs. 2.23 mSv in those aged 10-14 years; P<0.001) and at the RCH (2.44 mSv in those aged 0-3 years vs. 1.71 mSv in those aged 10-14 years; P<0.001). Each decreasing year of age was independently associated with a 0.06 mSv higher mean normalized ED (P<0.001). After adjusting for the age difference between the institutions, the mean normalized ED was 0.44 mSv lower at the RCH than at the TC across all ages (95% CI 0.31-0.58, P<0.001). A higher lifetime attributable risk of cancer was associated with younger age. CONCLUSION: The radiation dose from head CT in children as defined by the normalized ED was highest in the youngest children and varied significantly between institutions in this bi-institutional study. PMID- 19554323 TI - Congenital megalourethra. PMID- 19554324 TI - Molecular and biochemical characterization of a novel xylanase from the symbiotic Sphingobacterium sp. TN19. AB - A xylanase-encoding gene, designated xynA19, was cloned from Sphingobacterium sp. TN19--a symbiotic bacterium isolated from the gut of Batocera horsfieldi larvae- and expressed in Escherichia coli BL21 (DE3). The full-length xynA19 (1,155 bp in length) encodes a 384-residue polypeptide (XynA19) containing a predicted signal peptide of 24 residues and a catalytic domain belonging to glycosyl hydrolase family 10 (GH 10). The deduced amino acid sequence of XynA19 is most similar (53.1% identity) to an endo-1,4-beta-xylanase from Prevotella bryantii B(1)4. Phylogenetic analysis of GH 10 Bacteroidia xylanases indicated that GH 10 xylanases from Sphingobacteria were separated into two clusters, and XynA19 is more closely related to the xylanases of Bacteroidia from gut or rumen than to those of Flavobacteria and Sphingobacteria from other sources. Recombinant XynA19 (r-XynA19) showed apparent optimal activity at pH 6.5 and 45 degrees C. Compared with thermophilic and mesophilic counterparts, r-XynA19 was more active at low temperatures, retaining >65% of its maximum activity at 20-28 degrees C and approximately 40% even at 10 degrees C, and modeling indicated that XynA19 has fewer hydrogen bonds and salt bridges. These properties suggest that XynA19 has various potential applications, especially in aquaculture and the food industry. PMID- 19554325 TI - Continuous hydrolysis of 4-cyanopyridine by nitrilases from Fusarium solani O1 and Aspergillus niger K10. AB - The operational stabilities of nitrilases from Aspergillus niger K10 and Fusarium solani O1 were examined with 4-cyanopyridine as the substrate in continuous stirred membrane reactors (CSMRs). The former enzyme was fairly stable at 30 degrees C with a deactivation constant (k (d)) and enzyme half-life of 0.014 h( 1) and 50 h, respectively, but the latter exhibited an even higher stability characterized by k (d) = 0.008 h(-1) and half-life of 87 h at 40 degrees C. Another advantage of this enzyme was its high chemoselectivity, i.e., selective transformation of nitriles into carboxylic acids, while the amide formed a high ratio of A. niger K10 nitrilase product. High conversion rates (>90%) were maintained for about 52 h using the nitrilase from F. solani O1 immobilized in cross-linked enzyme aggregates (CLEAs). The purity of isonicotinic acid was increased from 98% to >99.9% by using two CSMRs connected in series, the first one containing the F. solani O1 nitrilase and the second the amidase from Rhodococcus erythropolis A4 (both enzymes as CLEAs), the amidase hydrolyzing the by-product isonicotinamide. PMID- 19554326 TI - Crush extremity fractures associated with the 2008 Sichuan earthquake: anatomic sites, numbers and statuses evaluated with digital radiography and multidetector computed tomography. AB - OBJECTIVE: To investigate features of crush extremity fractures associated with massive earthquake on digital radiography (DR) and multidetector computed tomography (MDCT). MATERIALS AND METHODS: Six hundred and twenty-three consecutive patients with clinically confirmed crush extremity fractures arising from the 2008 Sichuan earthquake were enrolled into our study. Six hundred and eleven patients with suggested extremity fractures underwent DR, and 12 patients with possible knee fractures underwent MDCT. Image data were retrospectively reviewed, with the focus on anatomic sites, numbers, and status of the fractures. RESULTS: Extremity fractures occurred in lower extremities in 428 patients, upper extremities in 151, and both lower and upper extremities in 44. Lower extremity fractures were more common than upper extremity fractures (P < 0.05), and the commonly involved bones were the tibia and fibula in 141 patients, femur in 102, tibia in 52, and fibula in 40. According to the numbers of bones involved, multiple bone fractures occurred in 336 patients and included lower extremity fractures in 231, upper extremity fractures in 61, and both lower and upper fractures in 44. Multiple fractures in lower extremities were seen more often than in upper extremities (P < 0.05). As for status of the extremity fractures, comminuted fractures occurred in 324 patients and included lower extremity fractures in 248, upper extremity fractures in 51, and both lower and upper extremity fractures in 25. Comminuted fractures were more common in lower extremities than in upper extremities (P < 0.05). CONCLUSION: Multiple and comminuted fractures, predominantly in the lower extremities, could be considered as features of crush extremity fractures associated with the massive Sichuan earthquake. PMID- 19554327 TI - Evaluation of early tissue reactions after lumbar intertransverse process fusion using CT in a rabbit. AB - OBJECTIVE: The objective of the study was to evaluate tissue reactions such as bone genesis, cartilage genesis and graft materials in the early phase of lumbar intertransverse process fusion in a rabbit model using computed tomography (CT) imaging with CT intensity (Hounsfield units) measurement, and to compare these data with histological results. MATERIALS AND METHODS: Lumbar intertransverse process fusion was performed on 18 rabbits. Four graft materials were used: autograft bone (n = 3); collagen membrane soaked with recombinant human bone morphogenetic protein-2 (rhBMP-2) (n = 5); granular calcium phosphate (n = 5); and granular calcium phosphate coated with rhBMP-2 (n = 5). All rabbits were euthanized 3 weeks post-operatively and lumbar spines were removed for CT imaging and histological examination. RESULTS: Computed tomography imaging demonstrated that each fusion mass component had the appropriate CT intensity range. CT also showed the different distributions and intensities of bone genesis in the fusion masses between the groups. Each component of tissue reactions was identified successfully on CT images using the CT intensity difference. Using CT color mapping, these observations could be easily visualized, and the results correlated well with histological findings. CONCLUSIONS: The use of CT intensity is an effective approach for observing and comparing early tissue reactions such as newly synthesized bone, newly synthesized cartilage, and graft materials after lumbar intertransverse process fusion in a rabbit model. PMID- 19554329 TI - Opening wedge osteotomy of the first cuneiform for the treatment of hallux valgus. AB - We performed an opening wedge osteotomy of the first cuneiform for the correction of all degrees of hallux valgus deformities. A wedge-shaped graft maintained the open wedge osteotomy and decreased the metatarsocuneiform joint inclination to correct the main angles of the hallux valgus. This procedure was performed for 101 feet of 63 patients (two men and 61 women). Fifteen feet benefited from a shortening of P1. The average age was 42 (16 to 84) years with a mean follow-up of 7.7 (1.5 to 14.8) years. We evaluated the M1-P1 and the M1-M2 angles, the joint congruency and the position of the lateral sesamoid. The clinical examination was graded by the AOFAS score which includes comprehensive assessment of pain, function and alignment and the subjective assessment of patients. The clinical AOFAS score improved from 35.24 points preoperatively to 86.36 postoperatively and from 33.25 to 88.03 points in severe deformity. The subjective rating was 44.44% excellent, 41.27% good, 9.52% fair and 4.76% poor. The M1-P1 angle improved from 29.38 degrees to 16.28 degrees and the M1-M2 angle from 14.36 degrees to 10.34 degrees . In the 42 severe feet deformity, these angles improved respectively from 45.58 degrees to 19.58 degrees and from 18.51 degrees to 11.16 degrees . This technique allowed an accurate correction of the main angles of hallux valgus with different degrees of deformity and avoided the complications related to different types of osteotomies achieved along the first metatarsal. After a long follow-up, we demonstrated a durable result while 86% of patients proved excellent and good results according to the AOFAS score. PMID- 19554328 TI - Distal tibia fractures: management and complications of 101 cases. AB - Distal tibia fractures are complex injuries with a high complication rate. In this retrospective and multicentre study we attempted to detail complications and outcomes of this type of injury in order to determine predictive factors of poor results. Between 2002 and 2004, 104 patients were admitted for 105 distal tibia fractures. One hundred patients (101 fractures) were reviewed with an average follow-up of 19 months (range, 12-46). Internal fixation, external fixation, limited internal fixation (K-wires or screws), intramedullary nailing and conservative treatment were used. Outcome parameters included occurrence of complications, radiographic analysis, evaluation of the American Orthopaedic Foot and Ankle Society (AOFAS) ankle score and measures of the ankle range of motion. The average functional score was 76 points (range, 30-100 points), and complications occurred in 30 patients. Predictive factors of poor results were fracture severity, complications, malunion and the use of external fixation. We believe that external fixation must be reserved for trauma with severe skin injury, as a temporary solution in a two-staged protocol. For other cases, we recommend ORIF with early mobilisation. PMID- 19554330 TI - Effects of bevacizumab on plasma concentration of irinotecan and its metabolites in advanced colorectal cancer patients receiving FOLFIRI with bevacizumab as second-line chemotherapy. AB - PURPOSE: Bevacizumab (BV) prolongs the survival of colorectal cancer patients when combined with irinotecan (CPT-11)-based regimens. In the AVF2107g study, the area under the curve (AUC) ratio for bolus CPT-11/5-fluorouracil (5 FU)/leucovorin (LV) (IFL) with the BV arm to bolus IFL with placebo indicated that SN-38 concentrations may have been increased in subjects receiving BV. However, the mechanism underlying such increase remains unclear, and the difference might be caused by an imbalance between the two arms and a possible inter-subject variability of CPT-11 metabolism. Within-subject comparisons were used to evaluate the effect of BV on advanced colorectal cancer patients when administered with the FOLFIRI regimen as second-line chemotherapy. METHODS: Ten advanced colorectal cancer patients received the FOLFIRI regimen every 2 weeks. At cycle 1, BV was administered following FOLFIRI administration to allow baseline pharmacokinetic (PK) analysis of CPT-11 and its metabolites. From cycle 2, BV was administered just before FOLFIRI administration. Plasma samples were collected under the same condition (at cycle 3). RESULTS: There were no significant differences in the Cmax and AUC0-infinity of CPT-11, SN-38, and SN 38G between cycle 1 (without BV) and cycle 3 (with BV). PK parameters of CPT-11, SN-38, and SN-38G were not significantly affected by BV. There were no significant differences in the changes in the AUC ratio of CPT-11 to SN-38 between cycles 1 and 3, as well as in the ratio of SN-38 to SN-38G. CONCLUSION: BV does not affect the plasma concentration of CPT-11 and its metabolites on FOLFIRI regimen. PMID- 19554331 TI - Structural and functional characterization of Delphinus delphis hemoglobin system. AB - Structural analysis of the hemoglobin (Hb) system of Delphinus delphis revealed a high globin multiplicity: HPLC-electrospray ionization-mass spectrometry (ESI-MS) analysis evidenced three major beta (beta1 16,022 Da, beta2 16,036 Da, beta3 16,036 Da, labeled according to their progressive elution times) and two major alpha globins (alpha1 15,345 Da, alpha2 15,329 Da). ESI-tandem mass and nucleotide sequence analyses showed that beta2 globin differs from beta1 for the substitution Val126 --> Leu, while beta3 globin differs from beta2 for the isobaric substitution Lys65 --> Gln. The alpha2 globin differs from the alpha1 for the substitution Ser15 --> Ala. Anion-exchange chromatography allowed the separation of two Hb fractions and HPLC-ESI-MS analysis revealed that the fraction with higher pI (HbI) contained beta1, beta2 and both the alpha globins, and the fraction with lower pI (HbII) contained beta3 and both the alpha globins. Both D. delphis Hb fractions displayed a lower intrinsic oxygen affinity, a decreased effect of 2,3-BPG and a reduced cooperativity with respect to human HbA(0), with HbII showing the more pronounced differences. With respect to HbA(0), either the substitution Probeta5 --> Gly or the Probeta5 --> Ala is present in all the cetacean beta globins sequenced so far, and it has been hypothesized that position 5 of beta globins may have a role in the interaction with 2,3-BPG. Regarding the particularly lowered cooperativity of HbII, it is interesting to observe that the variant human HbA, characterized by the substitution Lysbeta65 --> Gln (HbJ-Cairo) has a decreased cooperativity with respect to HbA(0). PMID- 19554332 TI - Grade II atypical choroid plexus papilloma with normal karyotype. AB - PURPOSE: Cytogenetic studies of atypical choroid plexus papillomas (CPP) have been poorly described. In the present report, the cytogenetic investigation of an atypical CPP occurring in an infant is detailed. METHODS: CPP chromosome preparations were analyzed by giemsa-trypsin-banding (GTG-banding) and comparative genome hybridization (CGH). RESULTS: Conventional karyotype analysis of tumor culture showed a normal chromosome complement. The results were confirmed by CGH, showing normal hybridization patterns for the sample. CONCLUSIONS: To date, the few atypical CPPs described in the literature have shown disparate cytogenetic information. This is the first report of a normal chromosome complement in atypical CPP. The heterogenic genetic features observed in these small series may reflect the diverse genetic background of choroid plexus tumors in children. PMID- 19554333 TI - Neuropsychological sequelae and quality of life following treatment of posterior fossa ependymomas in children. AB - INTRODUCTION: Modern treatment of posterior fossa ependymomas in children has improved the overall survival, but as more children live longer, the neuropsychological problems that they endure as result of the disease and its treatment become more the focus of attention. MATERIAL: Early problems such as cerebellar mutism appear to be related to surgery and do not seem to be influenced in their incidence or outcome by the otherwise beneficial effect of oncological treatment. Indeed, the cause of cerebellar mutism is still elusive. While it was originally believed to be totally reversible, it is increasingly realized that it may be related to more complex delayed neuropsychological problems. Late problems such as the cerebellar cognitive affective syndrome are increasingly being recognized. The deleterious effect of both surgery and radiotherapy that is commonly administered afterwards is now well appreciated. CONCLUSION: Careful neuropsychological assessment of these children and early intervention will maximize chances for recovery and improvement of academic functions and quality of life issues. PMID- 19554334 TI - Ultrastructural evidence of ependymal differentiation in a genetically proven atypical teratoid/rhabdoid tumor. AB - INTRODUCTION: We describe a case of genetically proven atypical teratoid/rhabdoid tumor (ATRT), showing ultrastructural evidence of ependymal differentiation. Such differentiation has not been reported so far in ATRT. CONCLUSION: This finding supports the concept that ATRTs as the majority of central nervous system embryonal tumors may derive from an immature and pluripotent neuroectodermal cell capable of differentiating along multiple lineages. PMID- 19554335 TI - Abdominal posterior rectopexy with an omental pedicle for intractable rectal prolapse: a modified technique. AB - INTRODUCTION: Rectal prolapse is a relatively common paediatric surgical condition. It has a number of benign aetiologies. Management is usually centred on regulating bowel habits. Surgery is considered after the failure of medical treatment. Numerous surgical techniques have been described with a spectrum of results. MATERIALS AND METHODS: We adopted a limited abdominal approach to achieve a posterior rectopexy using an omental pedicle in intractable cases. This technique has not been performed in children previously. RESULTS: From 2005 to 2008 we have applied this technique on five patients with recurrent rectal prolapse which had failed to respond to medical treatment, injection sclerotherapy or perianal cercalage. One patient had solitary rectal ulcer syndrome, and was initially treated with a defunctioning colostomy, had a concomitant sigmoidectomy performed at the time of rectopexy. None of the patients had cystic fibrosis. There were three females and two males, with a mean age of 9.6 years (4.7-14.0). No operative complications were encountered. The mean hospital stay was 5.4 days (3-8). None of the patients experienced recurrence at a mean of 2.1 years (0.2-2.8) follow up. The cosmetic result was regarded as satisfactory by all patients. CONCLUSION: This early experience with abdominal posterior rectopexy using an omental pedicle graft is encouraging. This technique does not involve the use of synthetic material and hence the risk of infection is low. PMID- 19554336 TI - [New aspects in the area of spondyloarthritis. Report on the 6(th) International Congress on Spondyloarthropathies]. AB - The concept of genetic principles and the pathophysiologic relationship in the course of spondyloarthropathies, especially in ankylosing spondylitis, has been strengthened significantly by a body of recent scientific data. These novel aspects have been presented at the 6(th) International Congress on Spondyloarthropathies in Ghent (October, 2(nd) - 4(th) 2008). PMID- 19554337 TI - Type and frequency of IDH1 and IDH2 mutations are related to astrocytic and oligodendroglial differentiation and age: a study of 1,010 diffuse gliomas. AB - Somatic mutations in the IDH1 gene encoding cytosolic NADP+-dependent isocitrate dehydrogenase have been shown in the majority of astrocytomas, oligodendrogliomas and oligoastrocytomas of WHO grades II and III. IDH2 encoding mitochondrial NADP+ dependent isocitrate dehydrogenase is also mutated in these tumors, albeit at much lower frequencies. Preliminary data suggest an importance of IDH1 mutation for prognosis showing that patients with anaplastic astrocytomas, oligodendrogliomas and oligoastrocytomas harboring IDH1 mutations seem to fare much better than patients without this mutation in their tumors. To determine mutation types and their frequencies, we examined 1,010 diffuse gliomas. We detected 716 IDH1 mutations and 31 IDH2 mutations. We found 165 IDH1 (72.7%) and 2 IDH2 mutations (0.9%) in 227 diffuse astrocytomas WHO grade II, 146 IDH1 (64.0%) and 2 IDH2 mutations (0.9%) in 228 anaplastic astrocytomas WHO grade III, 105 IDH1 (82.0%) and 6 IDH2 mutations (4.7%) in 128 oligodendrogliomas WHO grade II, 121 IDH1 (69.5%) and 9 IDH2 mutations (5.2%) in 174 anaplastic oligodendrogliomas WHO grade III, 62 IDH1 (81.6%) and 1 IDH2 mutations (1.3%) in 76 oligoastrocytomas WHO grade II and 117 IDH1 (66.1%) and 11 IDH2 mutations (6.2%) in 177 anaplastic oligoastrocytomas WHO grade III. We report on an inverse association of IDH1 and IDH2 mutations in these gliomas and a non-random distribution of the mutation types within the tumor entities. IDH1 mutations of the R132C type are strongly associated with astrocytoma, while IDH2 mutations predominantly occur in oligodendroglial tumors. In addition, patients with anaplastic glioma harboring IDH1 mutations were on average 6 years younger than those without these alterations. PMID- 19554338 TI - Screw fixation of the radial head: radiological assessment of the proximal radio ulnar joint and average radial head diameter-an anatomic study. AB - INTRODUCTION: Anatomical study of 11 radii heads was done. METHOD: A screw which is 2 mm longer than radial head diameter was used. Five views (AP with the forearm in supination, neutral rotation, pronation and LAT view the arm in supination and neutral rotation) were taken. In all cases the screw penetration was detected in one or two of the anteroposterior views. The lateral views were not sensitive to detect the screw penetration. For intraoperative assessment of screw length, the anteroposterior views in different position of rotation is sensitive method to detect penetration of the open reduction of head radius. Anteroposterior and lateral view with the arm in same position can miss some case of screw penetration. The diameter of the radial head was assessed. RESULT: A difference in the radial head diameter between males and females was noticed. This can be reflected on the screw diameter. PMID- 19554339 TI - Sentinel lymph node biopsy in two Burgundy districts: prospective multicentric study on 528 breast cancers during the year 2005. AB - OBJECTIVES: Our purpose was to assess development of sentinel lymph node biopsy (SLNB) in two Burgundy districts during the year 2005. METHODS: All women undergoing breast surgery as primary care between 1 January 2005 and 1 January 2006 were eligible for inclusion. Eleven surgeons from five different breast treatment centres took part in this prospective multicentric study. As our objective was to evaluate practices, patients were not randomized and surgeons were free to choose treatment patterns. RESULTS: The 528 enrolled cases account for 90% of all new breast cancers in 2005 in Cote d'Or and Saone et Loire. Half of these patients (286) fulfilled requirements for SLNB. The others (242) had primary full axillary clearance (AC). Four of our five centres offer double detection of sentinel lymph nodes as well as intraoperative pathology examination. Most tumours were invasive ductal carcinomas, with an average size of 12 mm in the SLNB group (T1C) and 22 mm in the AC group (T2). Two or three lymph nodes were removed during each SLNB procedure. Whereas most SLNB studies report around 25% positive nodes, we barely recorded 18.5% (53 of our 256 patients). Moreover, 2/3 of these node-positive patients had optimal care since additional axillary clearance was done right away. CONCLUSION: Sentinel lymph node biopsy has become routine practice in our Burgundy area. It is mainly dedicated to early stage breast cancer with limited metastatic risk. Our surgeons follow the most recent guidelines and indications are the same regardless of treatment centre. PMID- 19554340 TI - Computed tomographic angiography in diagnosis and management of placental polyp with neovascularization. AB - OBJECTIVE: To evaluate the neovascularization in placental polyp tissue by computed tomographic angiography and to determine the need for uterine artery embolization before hysteroscopic resection. STUDY DESIGN: Seventeen consecutive women with suspected placental polyp were enrolled in this retrospective study. Neovascularization in placental polyp tissue was assessed by computed tomographic angiography. Cases with neovascularization were treated by hysteroscopic resection with preoperative uterine artery embolization, while cases without neovascularization were treated by hysteroscopic resection alone. RESULTS: Of 17 patients with suspected placental polyp after abortion or parturition, nine patients were diagnosed to have placental polyp with prominent neovascularization by computed tomographic angiography, and were treated by uterine artery embolization followed by hysteroscopic resection. Two patients subsequently conceived after conservative management. CONCLUSIONS: After precise evaluation of neovascularization by computed tomographic angiography, hysteroscopic resection with preoperative uterine artery embolization is an effective minimally invasive procedure to conservatively treat placental polyp with prominent neovascularization. PMID- 19554341 TI - Is the current concept of recurrent ovarian carcinoma as a chronic disease also applicable in platinum resistant patients? AB - PURPOSE: The treatment of recurrent ovarian carcinoma (ROC) has become increasingly oriented according to the therapy principles of a chronic disease. We evaluated whether it is justifiable to also apply this concept to the treatment of platinum resistant patients with their known poor prognosis and short overall survival (OS). METHODS: We analyzed the overall courses of 85 unselected ROC patients and defined the following groups: A, platinum resistant patients (n=39); subgroup A.1, those who received no or at maximum one line of palliative chemotherapy (n=15, 38.5%); subgroup A.2, those who received>or=two therapy lines (n=24, 61.5%); B, platinum sensitive patients, n=46. RESULTS: Group A had significantly lower OS than group B (median: 16 vs. 25 months; p=0.019). Group A.1 had significantly worse outcome compared to group A.2 (median: 5 vs. 21.5 months; p<0.001). The comparison between study group A.2 and group B showed comparable survival rates (p=0.738). Considering only the patients who had completed treatment courses, the median number of therapy lines administered was higher in group A.2 than in group B (4 vs. 3; p=0.008). CONCLUSIONS: There is not only the known dichotomy between platinum sensitive and resistant ROC patients, but rather also within the platinum resistant subgroup itself. There is a considerably large subgroup of platinum resistant patients who will subsequently enter a phase where multiple treatment programs will be considered and administered. These patients have similar survival rates compared to those from the platinum sensitive patient group and the therapy principles of a chronic disease are applicable. PMID- 19554343 TI - The role of human beta defensins 2 and 3 in the second trimester amniotic fluid in predicting preterm labor and premature rupture of membranes. AB - AIM: Human beta defensins 2 (HBD2) and 3 (HBD3) are peptides expressed in the amnion and chorion. This is a matched case control study conducted in our Department to determine whether second trimester amniotic fluid HBD2 and HBD3 concentrations measured at the time of genetic amniocentesis could be potential markers of preterm labor prediction. METHODS: Amniotic fluid HBD2 and HBD3 were determined by an enzyme-linked immunosorbent assay (ELISA) Women with preterm labor were defined as cases (N=41) while for each case a woman matched for age delivering at term served as control (N=41). Subgroup analysis was conducted to examine possible associations of HBD2 and HBD3 in cases of premature rupture of membranes. Nineteen women with preterm labor and premature rupture of membranes were defined as cases while for every case a woman matched for maternal age delivering at term served as control (N1=19). Results were presented as odds ratios (OR) and 95% confidence intervals. Statistical analysis used STATA 8.2 and SPSS 11.5 edition. A P-value of <0.05 was considered statistically significant. RESULTS: Amniotic fluid concentrations of HBD2 at the time of genetic amniocentesis were positively associated with preterm premature rupture of membranes (P=0.028), but not with preterm labour. No association of HBD3 and preterm birth was documented. CONCLUSION: Second trimester amniotic fluid HBD2 might be a predictor of premature rupture of membranes. PMID- 19554342 TI - Pregnancy outcome of patients with dermoid and other benign ovarian cysts. AB - OBJECTIVE: To investigate pregnancy outcome of patients with dermoid and other benign ovarian cysts. METHODS: A population-based study comparing all pregnancies of women with and without benign ovarian cysts was conducted. Deliveries occurred during the years 1988-2007 at Soroka University Medical Center. A multivariate logistic regression model, with backward elimination, was constructed to find independent risk factors associated with benign ovarian cysts. RESULTS: During the study period there were 212,114 deliveries, of which 93 occurred in patients with benign ovarian cysts. Most of the lesions were benign cyatadenoma (41.9%), 36.7% were dermoid cyst, and 11.8% were adenofibroma, mostly diagnosed during cesarean delivery (76.3%). Others (12.9%) were diagnosed during pregnancy by ultrasonography and the remaining 10.8% were diagnosed before pregnancy. The mean diameter at diagnosis was 9.05+/-7.6 cm for cystadenoma, 6.09+/-3.0 cm for dermoid cyst and 4.55+/-4.1 cm for adenofibroma. Only 3 cases of ovarian torsion were noted (3.2%), and 15 cases of hospitalization due to abdominal pain (16.2%). The following conditions were significantly associated with benign ovarian cysts: hypertensive disorder [odds ratio (OR) 3.05; 95% confidence interval (CI) 1.87 4.97], and maternal age (OR 1.04; 95% CI 1.01-1.07). Ovarian dermoid cyst was significantly associated with fertility treatments (8.6 vs. 2.4% OR=3.75; 95% CI 1.1-12.2; P=0.019). In addition, after controlling for maternal age using a multivariate analysis, fertility treatments remained significantly associated with ovarian dermoid. No significant differences were noted between the groups regarding perinatal outcomes such as birth weight, low birth weight, congenital malformations, low Apgar scores, or perinatal mortality. CONCLUSION: The course of pregnancy of patients with dermoid and other benign ovarian cysts, including perinatal outcomes, is favorable. The cysts should be managed conservatively if possible with routine ultrasound follow up during the pregnancy since complications are extremely rare. PMID- 19554344 TI - The effect of topical ciprofloxacin and steroid-containing ear drops for chronic suppurative otitis media on the internal ear. AB - Aim of the study is to evaluate the effects of topical ciprofloxacin and prednisolone-containing ear drops for chronic suppurative otitis media on hearing threshold, cochlear reserve and cochlea morphology in healthy subjects and to determine the proper drug dose. Sixty-eight guinea pigs, all of which had healthy hearing, were used for the study. The first group (n = 30) was administered ciprofloxacin three times a day, the second group (n = 30) was administered prednisolone three times a day and the third group (n = 8) was administered sterile distilled water three times a day. The therapies lasted for 7 days and were administered intratympanically. The first group and second group were divided into three sub-groups of ten subjects. The first sub-group (n = 10) was administered an equivalent dose per kilogram as in humans, the second sub-group (n = 10) was administered one-third of the human-equivalent dose and the third sub-group (n = 10) was administered tenfold the human-equivalent dose. All subjects underwent brainstem evoked response audiometry (BERA) and distortion product otoacoustic emission (DPOAE) testing on the seventh and twenty-first days following the therapy. Following the tests, two subjects from each group were decapitated and examined under electron microscope. BERA and DPOAE testing results of the sub-group (n = 10) which was administered tenfold the human equivalent dose were significantly different from the control group and other groups (P < 0.05). According to electron microscopic examination of the cochlea, the group which was administered a tenfold human-equivalent dose of intratympanic ciprofloxacin and prednisolone showed atrophy in cells and degenerations in cilia. This case was statistically significant when compared with the control group and other groups (P < 0.05). Ciprofloxacin and prednisolone applied at a human-equivalent dose per kilogram did not affect the hearing and cochlear histology of subjects. PMID- 19554345 TI - Associations between two job stress models and measures of salivary cortisol. AB - PURPOSE: To investigate the association between two job stress models-the job demand-control model and the effort-reward imbalance model-and repeated measures of salivary cortisol among male and female call-centre operators. METHODS: Daily cortisol profiles consisting of seven time points were measured across two workdays and one leisure day to determine the cortisol awakening response and the cortisol output in the day in 104 volunteers. The employees completed two self administered questionnaire--the Karasek's demand-control questionnaire and the Siegrist's effort-reward imbalance questionnaire-to assess psychosocial hazards at work. The relations between the perceived workload measures and salivary cortisol levels were analyzed by means of generalized estimating equations method after adjusting for potential confounders (gender, age, educational level, marital status, morning awakening time, sleep duration and quality, weekdays, work schedule, adherence to sampling procedure). RESULTS: The total cortisol amount excreted in the awakening period was positively associated with the job strain measures (high strain vs. low strain: 1.4 (2.4-0.3) nmol/l). In contrast, individuals scoring higher in effort-reward imbalance at work had both lower cortisol awakening response (high imbalance vs. low imbalance: -0.7 (-1.3 to 0.2) nmol/l) and lower diurnal secretory activity (-9.2 (-17.7 to -0.7) nmol/l). Gender, weekday and adherence to sampling schedule significantly influenced the cortisol excretion in the morning period. CONCLUSIONS: Our results indicate that the two work stress models differentially affect salivary cortisol output. This finding suggests that combining the information from two complementary job stress models results in improved knowledge on the psychobiological correlates of the psychosocial work environment. PMID- 19554346 TI - Effects of short-term endurance exercise training on vascular function in young males. AB - We investigated effects of 6 days of endurance exercise training [cycling at 65% of peak oxygen consumption (VO(2peak)) for 2 h a day on six consecutive days] on vascular function in young males. Measures of VO(2peak), arterial stiffness, calf vascular conductance and heart rate variability were obtained pre- and post training. Indices of arterial stiffness were obtained by applanation tonometry to determine aortic augmentation index normalized to a heart rate of 75 bpm (AI(x) at 75 bpm), and central and peripheral pulse wave velocity (CPWV, PPWV). Resting and maximal calf vascular conductances were calculated from concurrent measures of blood pressure and calf blood flow using venous occlusion strain-gauge plethysmography. Time and frequency domain measures of heart rate variability were obtained from recording R-R intervals during supine and standing conditions. Both CPWV (5.9 +/- 0.8 vs. 5.4 +/- 0.8 m/s) and PPWV (9.7 +/- 0.8 vs. 8.9 +/- 1.3 m/s) were reduced following the training program. No significant changes were observed in AI(x) at 75 bpm, vascular conductance, heart rate variability or VO(2peak). These data indicate that changes in arterial stiffness independent of changes in heart rate variability or vascular conductance can be achieved in healthy young males following only 6 days of intense endurance exercise. PMID- 19554348 TI - Enhanced AGAMOUS expression in the centre of the Arabidopsis flower causes ectopic expression over its outer expression boundaries. AB - Spatial regulation of C-function genes controlling reproductive organ identity in the centre of the flower can be achieved by adjusting the level of their expression within the genuine central expression domain in Antirrhinum and Petunia. Loss of this control in mutants is revealed by enhanced C-gene expression in the centre and by lateral expansion of the C-domain. In order to test whether the level of central C-gene expression and hence the principle of 'regulation by tuning' also applies to spatial regulation of the C-function gene AGAMOUS (AG) in Arabidopsis, we generated transgenic plants with enhanced central AG expression by using stem cell-specific CLAVATA3 (CLV3) regulatory sequences to drive transcription of the AG cDNA. The youngest terminal flowers on inflorescences of CLV3::AG plants displayed homeotic features in their outer whorls indicating ectopic AG expression. Dependence of the homeotic feature on the age of the plant is attributed to the known overall weakening of repressive mechanisms controlling AG. Monitoring AG with an AG-I::GUS reporter construct suggests ectopic AG expression in CLV3::AG flowers when AG in the inflorescence is still repressed, although in terminating inflorescence meristems, AG expression expands to all tissues. Supported by genetic tests, we conclude that upon enhanced central AG expression, the C-domain laterally expands necessitating tuning of the expression level of C-function genes in the wild type. The tuning mechanism in C-gene regulation in Arabidopsis is discussed as a late security switch that ensures wild-type C-domain control when other repressive mechanism starts to fade and fail. PMID- 19554347 TI - Surgical strategy for sporadic primary hyperparathyroidism an evidence-based approach to surgical strategy, patient selection, surgical access, and reoperations. AB - PURPOSE: Progress in parathyroid imaging has brought substantial changes in the surgical strategy to approach patients with sporadic primary hyperparathyroidism (pHPT). The present review is focused on the safety and efficacy of limited parathyroid exploration. MATERIALS AND METHODS: Review of the literature focused on studies dealing with unilateral (two-gland exploration) or selective parathyroidectomy (one-gland exploration) in selected patients with pHPT and on the classification of published reports according to the degree of evidence. RESULTS: Parathyroid exploration limited to a solitary parathyroid adenoma can be considered a minimally invasive procedure that can be performed by the minicervicotomy, video-assisted, or endoscopic approaches. In properly selected patients, it affords results comparable to those of four-gland bilateral exploration in terms of cure and recurrence. It causes less postoperative hypocalcemia. CONCLUSIONS: Selective parathyroidectomy is an option for patients with positive preoperative localization tests undergoing first-time surgery who have no family history of pHPT, no goiter for which surgical therapy is proposed, and are not on lithium therapy. PMID- 19554349 TI - Duplication of the Asymmetric Leaves1/Rough Sheath 2/Phantastica (ARP) gene precedes the explosive radiation of the Ruschioideae. AB - The Mesembryanthemoideae and Ruschioideae subfamilies are a major component of the Greater Cape Floristic Region in southern Africa. The Ruschioideae show an astonishing diversity of leaf shape and growth forms. Although 1,585 species are recognised within the morphologically diverse Ruschioideae, these species show minimal variation in plastid DNA sequence. We have investigated whether changes in selected leaf development transcription factors underpin the recent, rapid diversification of this large group of succulent plants. Degenerate primers designed to conserved regions of Asymmetric Leaves1/Rough Sheath 2/Phantastica (ARP) and the Class III HD-ZIP family of genes, were used to amplify sequences corresponding to these genes from several species within the Mesembryanthemoideae and Ruschioideae subfamilies. Two members of the Class III HD-ZIP family were identified in both the Mesembryanthemoideae and Ruschioideae, and were derived from an ancient gene duplication event that preceded the divergence of gymnosperms and angiosperms. While a single ARP orthologue was identified in the Mesembryanthemoideae, two paralogues, ARPa and ARPb, were identified in the Ruschioideae subfamily. ARPa was present in all species of Ruschioideae analysed in this study. ARPb has been lost from the Apatesieae and Dorotheantheae tribes, which form an early evolutionary branch from the Ruschieae tribe, as well as from selected species within the Ruschieae. The recent duplication and subsequent selected gene loss of the ARP transcription factor correlates with the rapid diversification of plant forms in the Ruschioideae. PMID- 19554350 TI - Changes in localization and expression levels of Shroom2 and spectrin contribute to variation in amphibian egg pigmentation patterns. AB - One contributing factor in the worldwide decline in amphibian populations is thought to be the exposure of eggs to UV light. Enrichment of pigment in the animal hemisphere of eggs laid in the sunlight defends against UV damage, but little is known about the cell biological mechanisms controlling such polarized pigment patterns. Even less is known about how such mechanisms were modified during evolution to achieve the array of amphibian egg pigment patterns. Here, we show that ectopic expression of the gamma-tubulin regulator, Shroom2, is sufficient to induce co-accumulation of pigment granules, spectrin, and dynactin in Xenopus blastomeres. Shroom2 and spectrin are enriched and co-localize specifically in the pigmented animal hemisphere of Xenopus eggs and blastulae. Moreover, Shroom2 messenger RNA (mRNA) is expressed maternally at high levels in Xenopus. In contrast to Xenopus, eggs and blastulae of Physalaemus pustulosus have very little surface pigmentation. Rather, we find that pigment is enriched in the perinuclear region of these embryos, where it co-localizes with spectrin. Moreover, maternal Shroom2 mRNA was barely detectable in Physaleamus, though zygotic levels were comparable to Xenopus. We therefore suggest that a Shroom2/spectrin/dynactin-based mechanism controls pigment localization in amphibian eggs and that variation in maternal Shroom2 mRNA levels accounts in part for variation in amphibian egg pigment patterns during evolution. PMID- 19554351 TI - Genetics and the brain: many pathways to enlightenment. PMID- 19554352 TI - Plant community responses to 5 years of simulated climate change in meadow and heath ecosystems at a subarctic-alpine site. AB - Climate change was simulated by increasing temperature and nutrient availability in an alpine landscape. We conducted a field experiment of BACI-design (before/after control/impact) running for five seasons in two alpine communities (heath and meadow) with the factors temperature (increase of ca. 1.5-3.0 degrees C) and nutrients (5 g N, 5 g P per m(2)) in a fully factorial design in northern Swedish Lapland. The response variables were abundances of plant species and functional types. Plant community responses to the experimental perturbations were investigated, and the responses of plant functional types were examined in comparison to responses at the species level. Nutrient addition, exclusively and in combination with enhanced temperature increase, exerted the most pronounced responses at the species-specific and community levels. The main responses to nutrient addition were increases in graminoids and forbs, whereas deciduous shrubs, evergreen shrubs, bryophytes, and lichens decreased. The two plant communities of heath or meadow showed different vegetation responses to the environmental treatments despite the fact that both communities were located on the same subarctic-alpine site. Furthermore, we showed that the abundance of forbs increased in response to the combined treatment of temperature and nutrient addition in the meadow plant community. Within a single-plant functional type, most species responded similarly to the enhanced treatments although there were exceptions, particularly in the moss and lichen functional types. Plant community structure showed BACI responses in that vegetation dominance relationships in the existing plant functional types changed to varying degrees in all plots, including control plots. Betula nana and lichens increased in the temperature increased enhancements and in control plots in the heath plant community during the treatment period. The increases in control plots were probably a response to the observed warming during the treatment period in the region. PMID- 19554353 TI - Quality of life, anxiety, and depression in Turkish colorectal cancer patients. AB - INTRODUCTION: The aim of the present study is to investigate variations in quality of life as a function of depression and anxiety scores of colorectal cancer patients with Beck depression and State-Trait Anxiety Inventory (STAI) scoring system. DISCUSSION: One hundred ten patients with colorectal cancer undergoing chemotherapy who presented to Dokuz Eylul University Faculty of Medicine, Department of Oncology between January 2004 and April 2007 were included in this study. The series of forms including the questions regarding the demographic characteristics of the patient, Turkish version of the Beck Depression Inventory (BDI), the Turkish version of the STAI, and European Organization for Research and Treatment of Cancer Quality of Life Questionnaire C30 (EORTC-QLQ-C30; version 3) were completed during face-to-face interviews by trained interviewers to determine the psychological status and quality of life of the patients. The mean Beck depression scores were 11.2 +/- 9.0 (range 0-44) and the mean STAI scores were 41.9 +/- 8.8 (range 22-71). Of the patients (Beck depression scores >or=17 points), 23.6% were determined as depressive. The EORTC QLQ-C30 function scales and global quality of life scores of the depressive patients (BDI >or= 17) were significantly lower than that of the nondepressive patients (BDI < 17). EORTC-QLQ-C30 symptom scale scores (excluding of the diarrhea) of the depressive patients were significantly higher than that of the nondepressive patients. The patients with low STAI scores (STAI < 45) had significantly higher EORTC-QLQ-C30 function scales and global quality of life scores than the patients with high STAI scores (STAI >or= 45). EORTC-QLQ-C30 symptom scale scores of the patients with high STAI scores were significantly higher than that of the patients with low STAI scores. In the present study, we detected that anxiety and depression were strongly associated with poor quality of life in Turkish colorectal cancer patients. PMID- 19554354 TI - "It makes you feel so full of life" LiveWell, a feasibility study of a personalised lifestyle programme for colorectal cancer survivors. AB - GOALS: The acceptability and feasibility of a 3-month personalised lifestyle (diet, exercise and weight management) intervention in overweight adults who had completed curative treatment for colorectal cancer were assessed by qualitative interviews, quality-of-life questionnaires and subjective and objective measures of diet and activity. MAIN RESULTS: Over a 4-month period, 28 of 37 (75%) patients met the inclusion criteria and 20 (71%) of the eligible patients agreed to participate in the study and 18 (90%) completed the 3-month study. Reported adherence related to tailored advice, personalised feedback and family support. Reported barriers included time following surgery, fatigue, having a stoma or chronic diarrhoea and conflicting advice from clinicians. A weight change of -1.2 (+/-4.4) kg was achieved overall and -4.1 (+/-3.7) kg in the ten who had lost weight. CONCLUSIONS: Colorectal cancer survivors will participate in a lifestyle change initiative. Interventions should be personalised to suit abilities, provide feedback on personal goals and encourage social support. Intervention timing and attaining greater support from clinicians should be explored prior to the development of an efficacy trial. PMID- 19554355 TI - Structural analysis and the effect of cyclo(His-Pro) dipeptide on neurotoxins--a dynamics and density functional theory study. AB - The switching propensity and maximum probability of occurrence of the side chain imidazole group in the dipeptide cyclo(His-Pro) (CHP) were studied by applying molecular dynamics simulations and density functional theory. The atomistic behaviour of CHP with the neurotoxins glutamate (E) and paraquat (Pq) were also explored; E and Pq engage in hydrogen bond formation with the diketopiperazine (DKP) ring of the dipeptide, with which E shows a profound interaction, as confirmed further by NH and CO stretching vibrational frequencies. The effect of CHP was found to be greater on E than on Pq neurotoxin. A ring puckering study indicated a twist boat conformation for the six-membered DKP ring. Molecular electrostatic potential (MESP) mapping was also used to explore the hydrogen bond interactions prevailing between the neurotoxins and the DKP ring. The results of this study reveal that the DKP ring of the dipeptide CHP can be expected to play a significant role in reducing effects such as oxidative stress and cell death caused by neurotoxins. PMID- 19554356 TI - Gene deletion of cytosolic ATP: citrate lyase leads to altered organic acid production in Aspergillus niger. AB - With the availability of the genome sequence of the filamentous fungus Aspergillus niger, the use of targeted genetic modifications has become feasible. This, together with the fact that A. niger is well established industrially, makes this fungus an attractive micro-organism for creating a cell factory platform for production of chemicals. Using molecular biology techniques, this study focused on metabolic engineering of A. niger to manipulate its organic acid production in the direction of succinic acid. The gene target for complete gene deletion was cytosolic ATP: citrate lyase (acl), which had previously been identified by using genome-scale stoichiometric metabolic model simulations. The acl gene was deleted using the bipartite gene-targeting method, and the mutant was characterized in batch cultivation. It was found that the succinic acid yield was increased threefold by deleting the acl gene. Additionally, the total amount of organic acids produced in the deletion strain was significantly increased. Genome-scale stoichiometric metabolic model predictions can be used for identifying gene targets. Deletion of the acl led to increased succinic acid production by A. niger. PMID- 19554357 TI - Threshold determination in sweep VEP and the effects of criterion. AB - In order to develop criteria for the range of data points used for regression line fitting in sweep visually evoked potential (sVEP), which would be objective, clearly specified and give good repeatability and validity, and in order to investigate the effect of luminance on sVEP measurement, visual acuity (VA) and contrast sensitivity (CS) were measured with sVEP in adults aged 17-30 years and children aged 6-8 years. Six to ten participants took part in each experiment. Five criteria (C0-C4) for fitting the regression line were implemented. Test retest repeatability and validity against psychophysical thresholds at three luminance levels were considered for thresholds and the number of acceptable readings. There were significant effects of criteria (repeated measures ANOVAs, P < 0.05). The criteria, C2 and C3 (based on the range over which the signal-to noise ratio >or=1), consistently gave better VA and CS, more viable readings, better agreement with psychophysical thresholds in adults and better repeatability than the other criteria. In the case of adults, C2 gave thresholds that were not significantly different from the psychophysical thresholds (P > 0.05). There was little effect of luminance over the 25-100 cd/m(2) range used. Overall, C2 performed the best and would be the criterion of choice, giving better repeatability, better validity and more viable plots. PMID- 19554358 TI - Efficacy of a modified mandibular advancement device for a totally edentulous patient with severe obstructive sleep apnea. AB - BACKGROUND: It has been asserted that the success rate of oral appliances was more satisfactory for mild to moderate obstructive sleep apnea (OSA) than severe ones; besides, there is a lack of literature about mandibular advancement device (MAD) application for edentulous patients with OSA. REPORT: This clinical case shows fabrication method and treatment efficacy of a modified MAD, which is aiming to displace bulky masseter muscles laterally, to provide more space for tongue on totally edentulous patient with severe OSA. PMID- 19554361 TI - Low-level laser therapy attenuates creatine kinase levels and apoptosis during forced swimming in rats. AB - Studies suggest that high-intensity physical exercise can cause damage to skeletal muscles, resulting in muscle soreness, fatigue, inflammatory processes and cell apoptosis. The aim of this study was to investigate the effects of low level laser therapy (LLLT) on a decrease in creatine kinase (CK) levels and cell apoptosis. Twenty male Wistar rats were randomly divided into two equal groups: group 1 (control), resistance swimming; group 2 (LLLT), resistance swimming with LLLT. They were subjected to a single application of indium gallium aluminum phosphide (InGaAlP) laser immediately following the exercise for 40 s at an output power of 100 mW, wavelength 660 nm and 133.3 J/cm(2). The groups were subdivided according to sample collection time: 24 h and 48 h. CK was measured before and both 24 h and 48 h after the test. Samples of the gastrocnemius muscle were processed to determine the presence of apoptosis using terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick end labeling. (There was a significant difference in CK levels between groups (P < 0.0001) as well as between the 24 h and 48 h levels in the control group, whereas there was no significant intra-group difference in the LLLT group at the same evaluation times. In the LLLT group there were 66.3 +/- 13.2 apoptotic cells after 24 h and 39.0 +/- 6.8 apoptotic cells after 48 h. The results suggest that LLLT influences the metabolic profile of animals subjected to fatigue by lowering serum levels of CK. This demonstrates that LLLT can act as a preventive tool against cell apoptosis experienced during high-intensity physical exercise. PMID- 19554363 TI - Glomus tumor in the stomach. AB - Gastric glomus tumor is rare and most commonly described as a solitary, well defined, submucosal lesion in the antrum with non-specific clinical manifestations. We report this case with representative image findings, correlate with clinical presentations and pathologic demonstrations that can help to early detect and distinguish it from other malignant tumors such as gastrointestinal stromal tumor. PMID- 19554360 TI - The HMG-CoA reductase gene and lipid and lipoprotein levels: the multi-ethnic study of atherosclerosis. AB - HMG-CoA reductase (HMGCR) is an enzyme involved in cholesterol synthesis. To investigate the contribution of the HMGCR gene to lipids and lipoprotein subfractions in different ethnicities, we performed an association study in the Multi-Ethnic Study of Atherosclerosis (MESA). In total, 2,444 MESA subjects [597 African-Americans (AA), 627 Chinese-Americans (CHA), 612 European-Americans (EA), and 608 Hispanic-Americans (HA)] without statin use were included. Participants had measurements of blood pressure, anthropometry, and fasting blood samples. Subjects were genotyped for 10 single nucleotide polymorphisms (SNPs). After excluding SNPs with minor allele frequency <5%, a single block was constructed. The most frequent haplotype was H1 (41-56%) in all ethnic groups except AA (H2a, 44.9%). Lower triglyceride level was associated with the H2a haplotype in AA and H2 in HA. In HA, H4 carriers had higher levels of triglyceride and small low density lipoprotein (s-LDL), and lower high-density lipoprotein cholesterol (HDL c), while carriers with H7 or H8 had associations with these traits in the opposite direction. No significant association was discovered in both CHA and EA. The total variation for triglyceride that could be explained by H2 alone was 2.6% in HA and 1.4% in AA. In conclusion, HMGCR gene variation is associated with multiple lipid/lipoprotein traits, especially with triglyceride, s-LDL, and HDL c. The impact of the genetic variance is modest and differs greatly among ethnicities. PMID- 19554362 TI - Cyanotoxins from black band disease of corals and from other coral reef environments. AB - Many cyanobacteria produce cyanotoxins, which has been well documented from freshwater environments but not investigated to the same extent in marine environments. Cyanobacteria are an obligate component of the polymicrobial disease of corals known as black band disease (BBD). Cyanotoxins were previously shown to be present in field samples of BBD and in a limited number of BBD cyanobacterial cultures. These toxins were suggested as one of the mechanisms contributing to BBD-associated coral tissue lysis and death. In this work, we tested nine cyanobacterial isolates from BBD and additionally nine isolated from non-BBD marine sources for their ability to produce toxins. The presence of toxins was determined using cell extracts of laboratory grown cyanobacterial cultures using ELISA and the PP2A assay. Based on these tests, it was shown that cyanobacterial toxins belonging to the microcystin/nodularin group were produced by cyanobacteria originating from both BBD and non-BBD sources. Several environmental factors that can be encountered in the highly dynamic microenvironment of BBD were tested for their effect on both cyanobacterial growth yield and rate of toxin production using two of the BBD isolates of the genera Leptolyngbya and Geitlerinema. While toxin production was the highest under mixotrophic conditions (light and glucose) for the Leptolyngbya isolate, it was highest under photoautotrophic conditions for the Geitlerinema isolate. Our results show that toxin production among marine cyanobacteria is more widespread than previously documented, and we present data showing three marine cyanobacterial genera (Phormidium, Pseudanabaena, and Spirulina) are newly identified as cyanotoxin producers. We also show that cyanotoxin production by BBD cyanobacteria can be affected by environmental factors that are present in the microenvironment associated with this coral disease. PMID- 19554364 TI - A novel nostril retainer designed by a patient. PMID- 19554365 TI - Upper-lip augmentation by graft of preseptal orbicularis oculi muscle through blepharoplasty. AB - BACKGROUND: Upper-lip augmentation is used to enhance a thin upper lip or correct lip deficiencies or senile hypotrophy. We describe an easy, effective, and reproducible technique. METHODS: We use two preseptal orbicularis oculi muscle grafts that provide a reliable option for soft-tissue upper-lip augmentation, with improved vertical lip height and lateral lip projection and reappearance of the Cupid's bow. Muscle grafts are harvested from a blepharoplasty done at the same time. RESULTS: The advantages of this procedure include the creation of an anatomically natural upper lip through preserving the continuity and function of the labial structure, good augmentation, no donor-site morbidity, no visible scars on the vermilion, and successful rejuvenation with the associated blepharoplasty. CONCLUSION: Both patients and surgeons were satisfied with the results because the muscle grafts produce a youthful appearance by adding natural, soft roundness and fullness to the upper lip without an artificial look or the use of synthetic material, providing long-term augmentation. PMID- 19554367 TI - Computed axial tomography-guided fixation of sacroiliac joint disruption: safety, outcomes, and results at 3-year follow-up. AB - Secondary to the progress in interventional imaging, new therapeutic options have been developed that decrease potential complications because they are minimally invasive and they decrease patient rehabilitation time. As a diagnostic modality, computed axial tomography (CAT) allows precise evaluation of the degree of sacroiliac reduction that must be performed. Moreover, the use of CAT enables easy positioning of screws across the sacroiliac joint, thus avoiding nerve and vascular damage. We report our clinical experience of 20 patients treated by CAT guided percutaneous fixation for posttraumatic unilateral sacroiliac disruption, including evaluation of our technique, its safety, and patient outcomes and long term results. All patients in this study had successful outcomes, which were judged according to how much pain they experienced and how quickly they resumed normal activity after the procedure. Twelve of 16 patients were able to return to work by postoperative month 2. One patient had degenerative sacroiliac joint syndrome (5%), which was confirmed 6 months after surgery by CAT scan. None of the patients showed radiologic or clinical evidence of instability of the sacroiliac joint or screw migration. Postoperative follow-up, performed at 1, 2, and 3 years in our rehabilitation department, showed stable results over time. All pain disappeared, without the need for medication, in 19 patients (95%). PMID- 19554368 TI - Determination of the efflux pump-mediated resistance prevalence in Pseudomonas aeruginosa, using an efflux pump inhibitor. AB - In gram negative bacteria, fluoroquinolone resistance is acquired by target mutations in topoisomerase genes or by reducing the permeation of drugs due to the increase in expression of endogenous multidrug efflux pumps that expel structurally unrelated antimicrobial agents. An ongoing challenge is searching for new inhibitory substances in order to block efflux pumps and restore the antibiotic drugs susceptibility. In this research, we sought to investigate the interplay between ciprofloxacin and an efflux pump inhibitor (EPI), phenyl alanine arginyl beta-naphtylamide (PAbetaN), to determine the prevalence of efflux pump overexpression in clinical isolates of Pseudomonas aeruginosa. Ciprofloxacin was tested at different concentrations (256-0.25 microg/ml) with a fixed concentration of PAbetaN (50 microg/ml). The isolates susceptibility profiles were analyzed by disc diffusion and agar dilution methods using 10 antibiotic discs and 4 powders. It was found that in the presence of PAbetaN, resistance to ciprofloxacin was inhibited obviously and MIC values were decreased. The comparison between subgroups of P. aeruginosa isolates with different resistance profiles indicates that efflux pump overexpression (EPO) is present in 35% of ciprofloxacin resistant isolates with no cross resistance and in variable frequencies among isolates showing cross resistance to other tested antibiotics: gentamicin (31%), ceftazidime (29%), and imipenem (18%). Altogether, these results imply that PAbetaN maybe effective to restore the fluoroquinolone drugs susceptibility in clinical treatment procedures. Results also show that increased use of a fluoroquinolone drug such as ciprofloxacin can affect the susceptibility of P. aeruginosa to other different antipseudomonal agents. PMID- 19554369 TI - Hospital cost categories of one-stage versus two-stage management of common bile duct stones. AB - BACKGROUND: In the era of cost-conscious healthcare, hospitals are focusing on costs. Analysis of hospital costs per cost category may provide indications for potential cost-saving measures in the management of common bile duct stones (CBDS) with gallbladder in situ. METHODS: Between October 2005 and September 2006, 53 consecutive patients suffering from CBDS underwent either a one-stage procedure [laparoscopic common bile duct exploration (LCBDE) with stone clearance and cholecystectomy (LCCE)] or a two-stage procedure [endoscopic retrograde cholangiopancreatography with sphincterotomy and stone clearance (ERCP/ERS) followed by LCCE]. Costs were defined in different cost categories for each activity centre and were linked to the individual patient via the "bill of activities". Only patients (n = 38) with an uneventful post-procedural course and with available cost data were considered for cost analysis. Total length of hospital stay (LOS) was 2 (0-6) days after one-stage and 8 (3-18) days after two stage procedure (p < 0.0001). RESULTS: Costs per patient were significantly (p < 0.0001) less after one-stage versus two-stage management, i.e. total hospital costs (euro2,636 versus euro4,608), hospitalisation costs (euro701 versus euro2,190), consumables/pharmacy (euro645 versus euro1,476) and para-medical personnel (euro1,035 versus euro1,860; p = 0.0002). Operation room (OR) costs were comparable for one-stage and two-stage management (euro1,278 versus euro1,232; p = 0.280). Total hospital costs during ERCP were euro2,648 (euro729 4,544), during LCCE without LCBDE were euro2,101 (euro1,033-4,269), and during LCCE with LCBDE were euro2,636 (euro1,176-4,235). CONCLUSION: In the management of patients with CBDS and gallbladder in situ a one-stage procedure is associated with significantly less costs as compared with a two-stage procedure. From the economical point of view these patients should preferably be treated via a one stage procedure as long as safety and efficacy of this approach are provided. PMID- 19554371 TI - Two regulatory mechanisms of monoterpenoid pheromone production in Ips spp. of bark beetles. AB - Bark beetles use aggregation pheromones to coordinate host colonization and mating. These monoterpenoid chemical signals are produced de novo in midgut cells via the mevalonate pathway, and pheromone production is induced when an adult beetle feeds on phloem of a host tree. In Ips pini, juvenile hormone (JH) III influences key regulatory enzymes along the mevalonate pathway that leads to pheromone production. In fact, topically applied JH III is sufficient to stimulate pheromone production in unfed males. In this study, we explore the influence of feeding and JH III treatment on pheromone production in male Ips confusus, the pinyon Ips. We also characterize the influence of feeding and JH III treatment on transcript levels and activity of three key enzymes involved in pheromone biosynthesis: 3-hydroxy-3-methylglutaryl-CoA (HMG) synthase (HMGS), HMG CoA reductase (HMGR) and geranyl diphosphate synthase (GPPS). We also extend the current understanding of the regulation of pheromone biosynthesis in I. pini, by measuring the influence of feeding and JHIII treatment on enzymatic activity of HMGS and GPPS. Feeding on host phloem alone strongly induces pheromone production in male I. confusus, while JH III treatment has no effect. However, feeding and JH III both significantly up-regulate mRNA levels of key mevalonate pathway genes. Feeding up-regulates these genes to a maximum at 32 h, whereas with JH III treatment, they are up-regulated at 4, 8, and 16 h, but return near to non treatment levels at 32 h. Feeding, but not JH III treatment, also increases the activity of all three enzymes in I. confusus, while both feeding or treatment with JH III increase HMGS and GPPS activity in I. pini. Our data suggest that pheromone production in Ips is not uniformly controlled by JH III and feeding may stimulate the release of some other regulatory factor, perhaps a brain hormone, required for pheromone production. PMID- 19554370 TI - Liver ablation techniques: a review. AB - BACKGROUND: Ablation techniques for unresectable liver carcinomas have evolved immensely since their introduction. Results of studies involving these techniques are restricted to reports of patient case series, which are often not presented in a standardised manner. This review aims to summarise the major studies in ablation technologies and present them in a way that may make comparison between the major modalities easier. METHODS: All major databases (Medline, Cochrane, Embase and Pubmed) were searched for studies using microwave, radiofrequency or cryoablation to treat unresectable liver tumours. Only studies with at least 30 patients and 3-year follow-up were included. Complication, recurrence and survival rates of all studies are summarised and presented. RESULTS AND CONCLUSION: It is difficult to compare ablation modalities, as probe design and energy sources have evolved rapidly over the last decade. Ablation offers an invaluable palliative option and in some cases it may offer rates of cure approaching that of surgical resection with lower morbidity and mortality. Perhaps the time has come, therefore, for prospective large-scale randomised control trials to take place comparing ablation modalities to each other and surgical resection. PMID- 19554372 TI - (2S,8Z)-2-butyroxy-8-heptadecene: major component of the sex pheromone of chrysanthemum gall midge, Rhopalomyia longicauda. AB - The sex pheromone of the chrysanthemum gall midge, Rhopalomyia longicauda (Diptera: Cecidomyiidae), the most important insect pest in commercial plantations of chrysanthemum, Dendranthema morifolium (Ramat.) Tzvel., in China, was identified, synthesized, and field-tested. Volatile chemicals from virgin females and males were collected on Porapak in China and sent to the United Kingdom for analysis. Coupled gas chromatographic-electroantennographic detection (GC-EAG) analysis of volatile collections from females revealed two compounds that elicited responses from antennae of males. These compounds were not present in collections from males. The major EAG-active compound was identified as 2 butyroxy-8-heptadecene by gas chromatographic (GC) retention indices, mass spectra, in both electron impact and chemical ionization modes, hydrogenation, epoxidation, and derivatization with dimethyldisulfide. The lesser EAG-active compound was identified as the corresponding alcohol. The ratio of butyrate to alcohol in the collections was 1:0.26. Racemic (Z)-8-heptadecen-2-ol and the corresponding butyrate ester were synthesized from (Z)-7-hexadecenyl acetate, and the synthetic compounds found to have identical GC retention indices and mass spectra to those of the natural, female-specific components. Analysis of the volatile collections on an enantioselective cyclodextrin GC column showed the natural pheromone contained (2S,8Z)-2-butyroxy-8-heptadecene. Field tests showed that rubber septa containing racemic (Z)-2-butyroxy-8-heptadecene were attractive to R. longicauda males. The (naturally occurring) S-enantiomer was equally as attractive as the racemate, while the R-enantiomer was not attractive to males, and did not inhibit the activity of the S-enantiomer. The attractiveness of the butyrate was significantly reduced by the presence of even small amounts of the corresponding alcohol. PMID- 19554373 TI - Comprehensive analysis of the clinical significance of inducing pluripotent stemness-related gene expression in colorectal cancer cells. AB - BACKGROUND: We previously determined that cancer stem-like cells may influence the susceptibility of colorectal cancer (CRC) cells to chemotherapeutic agents. Although Takahashi and Park identified a set of induced pluripotent stem cell (iPS)-related genes required for normal stem cell maintenance, the precise role of iPS-related gene expression in CRC pathogenesis remains to be determined. The purpose of this study was to clarify the clinical relevance of "stemness" regulating gene expression in CRC cases. MATERIALS AND METHODS: Cancer cells were excised from tissues of 79 CRC cases by laser microdissection (LMD), and quantitative RT-PCR was used to evaluate expression levels of the iPS-related genes c-MYC, SOX2, OCT3/4, LIN28, KLF4, and NANOG, and to identify any associations between their expression and clinicopathological CRC progression. RESULTS: We found that LIN28 expression is significantly associated with lymph node metastasis (p = 0.018) and Dukes stage (p = 0.0319). SOX2expression is also correlated with lymph node metastasis. Furthermore, the ten cases with Dukes D disease expressed significantly higher levels of SOX2transcript than the other 69 cases (p = 0.0136). In contrast, KLF4 expression was inversely related to Dukes stage. Expression of c-MYC, OCT3/4, and NANOG did not appear to have clinical relevance in CRC cases. CONCLUSION: The present analysis strongly suggests that altered expression of several iPS-related genes plays a role in CRC pathogenesis. PMID- 19554374 TI - Chemotherapy-associated hepatotoxicity: do we need to be concerned? AB - Curative resection of liver metastases from colorectal cancer is associated with improved overall survival rates, making complete resection the goal of therapy for many patients. The addition of newer, more active chemotherapeutic regimens has prolonged patient survival in those with advanced disease and increased the number of patients eligible for surgical therapy. However, concerns regarding chemotherapy-associated liver injury may have a negative impact on the ability to offer potentially curative therapy and may increase morbidity in some patients. Specific forms of liver injury have been associated with various chemotherapeutic regimens, including steatosis and steatohepatitis with prolonged fluorouracil and irinotecan therapy, and sinusoidal injury with oxaliplatin-based regimens. While the histologic association appears likely, the contribution of chemotherapy associated hepatotoxicity to postoperative morbidity remains ill defined. PMID- 19554375 TI - Controversies in the management of hepatic colorectal metastases. PMID- 19554376 TI - Liver resection for metastatic colorectal cancer in the presence of extrahepatic disease. AB - BACKGROUND: Hepatic resection for metastatic colorectal cancer (CRC) with concomitant extrahepatic disease (EHD) is controversial. Earlier reports of the results of liver resection for metastatic CRC identified patients with EHD as a group with poor outcomes, suggesting that the presence of EHD was an absolute contraindication to resection. This has recently been challenged in several reports due to advances in systemic chemotherapy, surgical technique, and patient selection. METHODS: This review was restricted to published data in the English language identified by searches of MEDLINE and Pubmed databases as well as reference lists of recent review articles on subjects of surgery for metastatic colorectal cancer. RESULTS: Five-year survival after resection is worse than patients with liver-only disease but approximates the survival rates seen in patients with resected liver-only metastases in the era prior to the use of modern chemotherapy. Recurrence occurs in the great majority of patients. CONCLUSIONS: At this time, there appears to be a role for surgery in highly selected patients with a single site of EHD amenable to complete resection. Unlike patients with liver-only disease, however, the goals of surgery must not be viewed as potentially curative. PMID- 19554377 TI - The role of preoperative chemotherapy in patients with resectable colorectal liver metastases. AB - BACKGROUND: Liver metastases develop in 40-50% of patients with colorectal cancer and represent the major cause of death in this disease. Surgical resection remains the only treatment procedure that can ensure long-term survival and provide cure when liver metastases can be totally resected with clear margins, when the primary cancer is controlled, and when there is no nonresectable extrahepatic disease. Five-year survival rate after surgical resection of colorectal metastases varies from 25% to 55%, but cancer relapse is observed in most patients. AIM: To review the potential benefits and disadvantages of neoadjuvant chemotherapy administered before surgery to patients with initially resectable metastases. RESULTS: European Organization for Research and Treatment of Cancer (EORTC) study 40983 has shown that neoadjuvant chemotherapy could reduce the risk of relapse by one-quarter, and allows to test the chemosensitivity of the cancer, to help to determine the appropriateness of further treatments, and to observe progressive disease, which contraindicates immediate surgery. Neoadjuvant chemotherapy can induce damage to the remnant liver. Oxaliplatin-based combination regimen is associated with increased risk of vascular lesions, whereas irinotecan-containing regimens have been associated with increased risks of steatosis and steatohepatitis. Analysis of EORTC study 40983 showed that administration of six cycles of neoadjuvant systemic chemotherapy with 5-fluorouracil, leucovorin, and oxaliplatin (FOLFOX) was associated with moderate increase of the risk of reversible complications after surgery, but mortality rate was below 1% and not increased. If patients are not overtreated, chemotherapy before surgery is well tolerated. The integration of novel targeted agents in combination with cytotoxic drugs is a promising way to improve outcome in patients with advanced colorectal cancer. Preliminary trials have shown that targeted agents combined with cytotoxic regimens can increase tumor response rates. Another impact of preoperative chemotherapy is that metastases that respond to treatment may no longer be visible on computed tomography (CT) scan or at surgery. Patients should be carefully monitored and receive surgery before metastases disappear. CONCLUSION: Treatment of most patients with liver metastases-those with resectable metastases as well as those with initially unresectable metastases-should start with chemotherapy. If drugs are well chosen and the duration of treatment is monitored with care during multidisciplinary meetings, benefits largely outweigh potential disadvantages. PMID- 19554378 TI - Outcomes of ablation versus resection for colorectal liver metastases: are we comparing apples with oranges? PMID- 19554381 TI - Ultrasound-guided central venous cannulation in bariatric patients. AB - BACKGROUND: Central venous catheterization may be difficult in morbidly obese patients because anatomic landmarks are often obscured. METHODS: We evaluated the efficacy and safety of ultrasound-guided central venous cannulation in 55 patients undergoing bariatric surgery. The usefulness of ultrasonic examination combined with intraatrial electrocardiogram as a diagnostic tool for catheter misplacement was studied. RESULTS: Preliminary ultrasound examination of the neck vessels demonstrated anatomical variations in the position of internal jugular vein in 19 cases and four unrecognized asymptomatic thromboses of the right internal jugular vein. Central venous catheterization was successful in all 55 patients, in 51 with single skin puncture, and in 42 with single vein puncture. In three cases in whom the catheter was misplaced, this was detected by bedside ultrasonic examination during the procedure and immediately corrected by real time echographic visualization. No arterial puncture, no hematoma, and no pneumothorax occurred in any patient. Successful catheter placement was also confirmed in all patients by post-operative chest X-ray. No evidence of infection or thrombosis subsequently was noted. CONCLUSIONS: The use of ultrasound guidance may increase the success rate and decrease the incidence of complications associated with central venous cannulation. The advantages of this approach is visualization of the anatomical structures at puncture site prior to skin puncture and the ability to track needle and guide-wire placement during the procedure. With its high accuracy in detecting catheter misplacement, bedside ultrasonic examination combined with intraatrial electrocardiogram may further decrease morbidity associated with misplaced central venous catheters. PMID- 19554383 TI - Limited quadricepsplasty for contracture during femoral lengthening. AB - Extension contracture of the knee is a common complication of femoral lengthening. Knee flexion exercises to stretch the contracture with physical therapy can be effective but take a prolonged amount of time to work and place increased stress across the patellofemoral joint. We developed a minimal-incision limited quadricepsplasty surgical technique to treat knee extension contracture secondary to femoral lengthening and retrospectively reviewed 16 patients treated with this procedure. The mean age of the patients was 23 years. Range of motion of the knee and quadriceps strength were recorded preoperatively, after femur lengthening but before additional surgery, after quadricepsplasty, and at each followup. The mean femoral lengthening performed was 4.4 cm. We compared range of motion and time to regain knee flexion with those of historical controls. The minimum followup after quadricepsplasty was 6 months (mean, 38 months; range, 6 84 months). The mean range of motion was 129 degrees preoperatively, 29 degrees after the distraction phase of femoral lengthening, and 108 degrees after limited quadricepsplasty, and at final followup, the mean knee flexion was 125 degrees . There were no major complications. Limited quadricepsplasty improved knee flexion after a knee extension contracture developed secondary to femoral lengthening. In comparison to historical controls who did not have quadricepsplasty, the patients with limited quadricepsplasty had quicker return of knee flexion, although there was no difference in knee flexion achieved ultimately. LEVEL OF EVIDENCE: Level III, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19554382 TI - Behavioral predictors of weight regain after bariatric surgery. AB - BACKGROUND: After bariatric surgery, a lifelong threat of weight regain remains. Behavior influences are believed to play a modulating role in this problem. Accordingly, we sought to identify these predictors in patients with extreme obesity after Roux-en-Y gastric bypass (RYGB). METHODS: In a large tertiary hospital with an established bariatric program, including a multidisciplinary outpatient center specializing in bariatric medicine, with two bariatric surgeons, we mailed a survey to 1,117 patients after RYGB. Of these, 203 (24.8%) were completed, returned, and suitable for analysis. Respondents were excluded if they were less than 1 year after RYGB. Baseline demographic history, preoperative Beck Depression Inventory (BDI), and Brief Symptom Inventory-18 scores were abstracted from the subjects' medical records; pre- and postoperative well-being scores were compared. RESULTS: Of the study population, mean age was 50.6 +/- 9.8 years, 147 (85%) were female, and 42 (18%) were male. Preoperative weight was 134.1 +/- 23.6 kg (295 +/- 52 lb) and 170.0 +/- 29.1 kg (374.0 +/- 64.0 lb) for females and males, respectively, p < 0.0001. The mean follow-up after bariatric surgery was 28.1 +/- 18.9 months. Overall, the mean pre- versus postoperative well-being scores improved from 3.7 to 4.2, on a five-point Likert scale, p = 0.001. A total of 160 of the 203 respondents (79%) reported some weight regain from the nadir. Of those who reported weight regain, 30 (15%) experienced significant regain defined as an increase of > or =15% from the nadir. Independent predictors of significant weight regain were increased food urges (odds ratios (OR) = 5.10, 95% CI 1.83-14.29, p = 0.002), severely decreased postoperative well-being (OR = 21.5, 95% CI 2.50-183.10, p < 0.0001), and concerns over alcohol or drug use (OR = 12.74, 95% CI 1.73-93.80, p = 0.01). Higher BDI scores were associated with lesser risk of significant weight regain (OR = 0.94 for each unit increase, 95% CI 0.91- 0.98, p = 0.001). Subjects who engaged in self-monitoring were less likely to regain any weight following bariatric surgery (OR = 0.54, 95% CI 0.30-0.98, p = 0.01). Although the frequency of postoperative follow-up visits was inversely related to weight regain, this variable was not statistically significant in the multivariate model. CONCLUSIONS: Predictors of significant postoperative weight regain after bariatric surgery include indicators of baseline increased food urges, decreased well-being, and concerns over addictive behaviors. Postoperative self-monitoring behaviors are strongly associated with freedom from regain. These data suggest that weight regain can be anticipated, in part, during the preoperative evaluation and potentially reduced with self-monitoring strategies after RYGB. PMID- 19554384 TI - Can porous tantalum be used to achieve ankle and subtalar arthrodesis?: a pilot study. AB - A structural graft often is needed to fill gaps during reconstructive procedures of the ankle and hindfoot. Autograft, the current gold standard, is limited in availability and configuration and is associated with donor-site morbidity in as much as 48%, whereas the alternative allograft carries risks of disease transmission and collapse. Trabecular metal (tantalum), with a healing rate similar to that of autograft, high stability, and no donor-site morbidity, has been used in surgery of the hip, knee, and spine. However, its use has not been documented in foot and ankle surgery. We retrospectively reviewed nine patients with complex foot and ankle arthrodeses using a tantalum spacer. Minimum followup was 1.9 years (average, 2 years; range, 1.9-2.4 years). Bone ingrowth into the tantalum was analyzed with micro-CT in three of the nine patients. All arthrodeses were fused clinically and radiographically at the 1- and 2 year followups and no complications occurred. The American Orthopaedic Foot and Ankle Society score increased from 32 to 74. The micro-CT showed bony trabeculae growing onto the tantalum. Our data suggest tantalum may be used as a structural graft option for ankle and subtalar arthrodesis. All nine of our patients achieved fusion and had no complications. Using tantalum obviated the need for harvesting of the iliac spine. LEVEL OF EVIDENCE: Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19554386 TI - Depressed affect is associated with poorer cardiovascular recovery in young women following a mental stressor. AB - BACKGROUND: Depressed mood has been prospectively associated with hypertension. Altered ANS function, as reflected in poor CV recovery, may be one mechanism that underlies this relationship. PURPOSE: The purpose of this study was to investigate the relationship between depressed mood and cardiovascular recovery following a standard mental stress task in healthy young women. METHODS: Depressed mood was assessed in 63 young women. Cardiovascular data were collected during a 5-min baseline period, 5-min public speaking stress task, and 15-min recovery period. RESULTS: Depressed mood accounted for 9.6% of the variation in HR reactivity (F(1,58) = 6.513, p = 0.013) and 4.5% of DBP recovery (F(1,58) = 4.538, p = 0.037). CONCLUSIONS: Greater depressed mood was associated with greater HR reactivity and poorer DBP recovery. This is the first study to directly investigate associations between depressed mood and CV recovery following mental stress. PMID- 19554385 TI - The epidemiology of revision total knee arthroplasty in the United States. AB - Understanding the cause of failure and type of revision total knee arthroplasty (TKA) procedures performed in the United States is essential in guiding research, implant design, and clinical decision making in TKA. We assessed the causes of failure and specific types of revision TKA procedures performed in the United States using newly implemented ICD-9-CM diagnosis and procedure codes related to revision TKA data from the Nationwide Inpatient Sample (NIS) database. Clinical, demographic, and economic data were reviewed and analyzed from 60,355 revision TKA procedures performed in the United States between October 1, 2005 and December 31, 2006. The most common causes of revision TKA were infection (25.2%) and implant loosening (16.1%), and the most common type of revision TKA procedure reported was all component revision (35.2%). Revision TKA procedures were most commonly performed in large, urban, nonteaching hospitals in Medicare patients ages 65 to 74. The average length of hospital stay (LOS) for all revision TKA procedures was 5.1 days, and the average total charges were $49,360. However, average LOS, average charges, and procedure frequencies varied considerably by census region, hospital type, and procedure performed. LEVEL OF EVIDENCE: Level II, economic and decision analysis. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19554387 TI - The treasury of the commons: making use of public gene expression resources to better characterize the molecular diversity of inhibitory interneurons in the cerebellar cortex. AB - We mined the Allen Mouse Brain Atlas for genes expressed in cerebellar cortical inhibitory interneurons that would allow identification and possibly distinction of these cells. We identified some 90 genes that are highly expressed in specific subsets of cerebellar cortical inhibitory interneurons or various combinations thereof. Four genes are exclusively expressed, within the cerebellar cortex, in molecular layer interneurons, and ten genes label exclusively inhibitory interneurons in the granule cell layer or subsets thereof. Differential expression of many of these genes in cells residing in the lower versus the upper molecular layer provides evidence that these cells, traditionally referred to as basket and stellate cells, are indeed molecularly distinct. Two genes could be identified as novel markers for unipolar brush cells. Intersection of these data with embryonic expression patterns as documented in the genepaint repository does not support a hierarchical model of cerebellar interneuron development, but may be more easily reconciled with the view that cerebellar inhibitory interneurons derive from a common precursor pool from which they are specified only late into their development. The novel markers identified here should prove useful for probing the timing and mechanisms supporting cerebellar cortical interneuron specification and diversification. PMID- 19554389 TI - Preoperative diagnosis and successful laparoscopic treatment of incarcerated obturator hernia. AB - Obturator hernia is rare and frequently delayed in surgical treatment and, thus, is associated with high morbidity and mortality. It is often found in thin, elderly, multiparous women. We reported a case of early diagnosis due to clinical alert and in-time computed tomography (CT). The patient received laparoscopic surgery and had good postoperative recovery. The regular use of CT for diagnosing nonspecific abdominal complaints associated with pain over the inner thigh, especially in thin, elderly, multiparous women, will result in increased preoperative detection of the obturator hernia. PMID- 19554388 TI - Oxidized low-density lipoprotein-induced matrix metalloproteinase-9 expression via PKC-delta/p42/p44 MAPK/Elk-1 cascade in brain astrocytes. AB - After ischemic injury to brain, disruption of the blood-brain barrier (BBB) raises the possibility of exposing the central nervous system (CNS) to oxidized low-density lipoprotein (oxLDL), a risk factor implicated in neurodegenerative diseases. Matrix metalloproteinases (MMPs), especially MMP-9, contribute to extracellular matrix (ECM) remodeling during the CNS diseases. However, the molecular mechanisms underlying oxLDL-induced MMP-9 expression in astrocytes remained unclear. Here, we reported that oxLDL induced MMP-9 expression via a PKC delta/p42/p44 MAPK-dependent Elk-1 activation in rat brain astrocyte (RBA)-1 cells, revealed by gelatin zymography, RT-PCR, and Western blotting analyses. These responses were attenuated by pretreatment with pharmacological inhibitors and transfection with dominant negative mutants. Moreover, Elk-1-mediated MMP-9 gene transcription was confirmed by transfection with an Elk-1 binding site mutated MMP-9 promoter construct (mt-Ets-MMP9), which blocked oxLDL-stimulated MMP-9 luciferase activity. Understanding the regulatory mechanisms by which oxLDL induced MMP-9 expression in astrocytes might provide a new therapeutic strategy of brain diseases. PMID- 19554390 TI - Urologic pelvic surgery following mesh hernia repair. AB - PURPOSE: Few and controversial reports have recently appeared on the role of previously performed surgery in the inguinal region using a prosthetic mesh and the ensuing difficulties encountered by urologists during radical retropubic prostatectomy. We analyzed our experience with various surgical urological procedures performed after prior low abdominal wall hernia repair with synthetic mesh. METHODS: We reviewed our database for all patients who underwent mesh repair of lower abdominal hernias and subsequent urologic surgery in our department between 2002 and 2008. Their perioperative parameters, complications, and postoperative outcomes were analyzed. RESULTS: Twenty-three patients (one female) underwent pelvic urologic surgery for benign and malignant disease after having undergone previous prosthetic hernia repair. The mean patient age was 75.3 years (range 58-91). The mean interval between hernia repair and pelvic urologic surgery was 3.8 years (range 1-7). Twenty-two patients underwent previous mesh inguinal hernia repair and one had prosthetic postoperative ventral hernia repair after a transabdominal hysterectomy. The urologic procedures included 16 open suprapubic prostatectomies, two radical cystoprostatectomies, one bladder augmentation, and four laparoscopic radical prostatectomies. Severe postoperative complications were abortion of surgery (n = 1), inability to perform lymph node dissection (n = 2), bleeding (n = 1), bladder injury (n = 2), and additional surgery (n = 3: mesh removal, transurethral prostatectomy, and transurethral fulguration of the prostatic fossa). CONCLUSIONS: Prior application of synthetic mesh during abdominal wall surgery creates difficulties during subsequent urological procedures and may dictate change in operative planning. Nevertheless, the surgery is feasible and should not be ruled out. PMID- 19554391 TI - The influence of hepatitis B DNA level and antiviral therapy on recurrence after initial curative treatment in patients with hepatocellular carcinoma. AB - BACKGROUND: Prediction and prevention of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) recurrence is an important clinical issue. We investigated whether HBV DNA level and antiviral therapy are associated with HCC recurrence. METHODS: This retrospective study involved 103 patients who underwent hepatic resection or radiofrequency ablation for initial HCC. Patients were divided into four groups. Thirty had high serum HBV DNA levels (>4 log(10) copies/mL) and had not received antiviral therapy (high virus group; HVG). Thirty four had low HBV DNA levels (< or =4 log(10) copies/mL) and had not received antiviral therapy (low virus group; LVG). Twenty received antiviral therapy after HCC developed (therapeutic group A, TG-A). Nineteen received antiviral therapy before HCC developed (therapeutic group B, TG-B). RESULTS: Cumulative HCC recurrence rates at 3 years in the HVG, LVG, TG-B, and TG-A were 71.1%, 42.2%, 42.3%, and 52.0%, respectively. Recurrence rates differed significantly between the HVG and LVG (P = 0.016) and between the HVG and TG-B (P = 0.008). Recurrence rate in the TG-A was marginally lower than in the HVG (P = 0.10). On multivariate analysis, high serum hepatitis B virus DNA levels (hazard ratio: HR 2.67; 95% CI 1.31-5.47; P = 0.007) and absence of antiviral therapy (HR 2.57; 95% CI 1.34 4.94; P = 0.005) were independent risk factors for hepatocellular carcinoma recurrence. CONCLUSION: HBV DNA level and antiviral therapy are associated with HCC recurrence. For patients with high HBV DNA levels, antiviral therapy before the development of HCC is important for prevention of recurrence. PMID- 19554392 TI - Local delivery of rolipram, a phosphodiesterase-4-specific inhibitor, augments bone morphogenetic protein-induced bone formation. AB - Recombinant human bone morphogenetic protein (rhBMP) is a promising therapeutic cytokine for the induction of bone formation, but a weak response in humans remains a major hurdle in its therapeutic application. We have previously reported an rhBMP-2-induced increase in the bone mass of mice receiving systemic rolipram, a specific inhibitor of phosphodiesterase-4. To overcome the side effects of systemic administration of rolipram, we examined the effects of its local release. Polyethylene glycol discs were used as a delivery system. The discs were impregnated with rhBMP-2 and rolipram and implanted into the dorsal muscle pouches in mice. Bone formation was assessed by measuring the bone mineral content (BMC) of the formed bone. First, to determine the optimal dose of rolipram, we added 0-5000 nmol rolipram and 5 microg rhBMP-2 to the pellets and found that 500 nmol rolipram was the most effective concentration for inducing bone formation after 4 weeks. Second, to examine the time course of bone formation, we implanted 5 microg rhBMP-2 with 0 or 500 nmol rolipram and killed mice 5, 7, 10, 14, or 21 days after implantation. Bone formation was accelerated in the rolipram group. Finally, to determine the rolipram-induced increase in the effect of BMP, BMC obtained after treatment with 5 microg rhBMP-2 and 500 nmol rolipram was compared with that obtained after treatment with 5-9 microg rhBMP-2 without rolipram, 4 weeks after implantation. The results indicated that 500 nmol rolipram enhanced the effect of rhBMP-2 by almost 1.5-fold. In summary, locally released rolipram enhanced the capacity of rhBMP-2 to induce bone formation, an effect previously reported with systemic administration. These findings may decrease the cost and increase the efficacy of rhBMP-2 treatment. PMID- 19554393 TI - Rheumatoid arthritis is linked to oral bacteria: etiological association. AB - The purpose of this review is to evaluate the association between rheumatoid arthritis (RA) and periodontopathic bacteria. Clinical studies of RA and periodontal disease have provided evidence for a significant association between the two disorders. Patients with long-standing active RA have a substantially increased frequency of periodontal disease compared with that among healthy subjects. High levels of oral anaerobic bacterial antibodies have been found in the serum and synovial fluid of RA patients. Porphyromonas gingivalis, Tannerella forsythensis, and Prevotella intermedia have been identified in RA synovial fluid. Ornidazole, levofloxacin, and clarithromycin are used in the treatment of infections caused by anaerobic bacteria. These antibiotics have been shown to be effective against RA. The evidence in this review indicates that oral bacteria directly associate with etiopathogenesis of RA. PMID- 19554395 TI - Curcumin prevents liver fibrosis by inducing apoptosis and suppressing activation of hepatic stellate cells. AB - This study was designed to investigate the prophylactic effects and the mechanisms of curcumin on liver fibrosis in rats. Liver fibrosis was induced in 72 Sprague Dawley rats by intraperitoneal injection of carbon tetrachloride. Rats were divided into control, liver fibrosis, high, medium, and low dose curcumin (200, 100, and 50 mg kg(-1), respectively), and colchicine (0.1 mg kg(-1)) groups. After 8 weeks of treatment, histopathological examination was performed on hepatic tissues, and liver fibrosis was graded. Hepatic stellate cells activity was examined by smooth muscle alpha-actin immunohistochemistry staining, and apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick-end labeling. The liver fibrosis score in the high, medium, and low dose curcumin group (5.79 +/- 1.80, 8.58 +/- 3.34, and 9.58 +/- 3.32, respectively) and the colchicine group (4.91 +/- 1.28) was significantly lower than in the fibrosis group (20.40 +/- 3.38, P < 0.01). The ratio of activated hepatic stellate cells in the three curcumin groups (0.97 +/- 0.69, 2.06 +/- 0.58, and 3.49 +/- 1.03, respectively) and the colchicine group (0.78 +/- 0.31) was significantly lower than in the fibrosis group (6.08 +/- 1.13, P < 0.05). The apoptosis index in the three curcumin groups (0.57 +/- 0.21, 0.37 +/- 0.22, and 0.34 +/- 0.21, respectively) was higher than in the fibrosis (0.09 +/- 0.09, P < 0.05) or the colchicine group (0.16 +/- 0.19, P < 0.05). Curcumin prevents carbon tetrachloride-induced liver fibrosis in rats. The prevention of liver fibrosis may be due to the inhibition of the activation of hepatic stellate cells and induction of their apoptosis. PMID- 19554396 TI - Tocilizumab is effective for pulmonary hypertension associated with multicentric Castleman's disease. AB - A 38-year-old man, diagnosed as having multicentric Castleman's disease (plasma cell type) in 1995, had been treated with melphalan and prednisolone or prednisolone alone, but there was no remarkable response. In 2002, he was admitted to our hospital with a chief complaint of increasing dyspnea on effort. Laboratory data showed high serum IgG (10050 mg/dl), interleukin-6 (37.9 ng/ml), and vascular endothelial growth factor (VEGF 1920 pg/ml) levels. In addition, serum viscosity was very high (6.0 cp). Electrocardiogram, echocardiogram, and cardiac catheterization demonstrated pulmonary hypertension (PH). There were no other demonstrable causes of PH suggesting that PH was due to hyperviscosity syndrome and high VEGF level. He was treated with plasmapheresis, resulting in a transient improvement of dyspnea. Then, he was given humanized anti-interleukin-6 receptor antibody (tocilizumab), which resulted in the dramatic improvement of dyspnea and PH a few weeks later. PH is a rare complication of MCD, and could be successfully treated with tocilizumab. PMID- 19554397 TI - A false positive for metastatic lymph nodes in the axillary region of a breast cancer patient following mastectomy. AB - Recent advanced imaging modalities such as positron emission tomography (PET) detect malignancies using 2-[18F]-fluoro-2-deoxy-D: -glucose (18-FDG) with high accuracy, and they contribute to decisions regarding diagnosis, staging, recurrence, and treatment response. Here, we report a case of false-positive metastatic lymph nodes that were diagnosed by PET/CT and ultrasonography in a 48 year-old breast cancer patient who had undergone mastectomy. The tumors, which were oval shaped and resembled lymph nodes, were detected by ultrasonography. PET/CT revealed high uptake of 18-FDG in the tumors. To investigate the proposed recurrence and to re-evaluate the biology of the recurrent tumors, a tumor was removed from the brachial plexus of the patient. Histological findings revealed it to be a schwannoma. All imaging modalities including PET/CT failed to distinguish benign tumors from metastatic lymph nodes in the brachial plexus. After resection of the schwannomas, the patient complained of a slight motor disorder of the second finger on the right hand. Hence, it is important to consider a false-positive case of lymph node metastasis in a breast cancer patient following mastectomy. PMID- 19554398 TI - Pathogenesis and clinical manifestations of chronic colonization by Pseudomonas aeruginosa and its biofilms in the airway tract. PMID- 19554399 TI - Usefulness of interferon-gamma release assays for diagnosing TB infection and problems with these assays. AB - The specificity of the tuberculin skin test (TST) in the diagnosis of tuberculosis infection is seriously compromised because of extensive use of the bacille Calmette-Guerin (BCG) vaccination. The interferon-gamma release assay (IGRA), a new diagnostic using Mycobacterium tuberculosis-specific antigens has been introduced in response to these needs. In this review, published findings on the performance of the QuantiFERON-TB (QFT), one of the IGRA formats, are summarized and discussed. In addition to its high specificity, the QFT has considerably high sensitivity, comparable with or superior to that of the TST, if applied to patients with active tuberculosis as a surrogate of latent tuberculosis infection. When applied to patients with immunosuppression, such as aging patients, or those with HIV infection, those with immunosuppressive drug therapies, or those with renal hemodialysis, QFT is shown to be more robust than the TST. As regards the dynamics of QFT responses to chemotherapy, there are many reports showing a decrease in responses during the treatment, which indicates the possibility that QFT could be used as a tool for monitoring the progress of treatment. However, there are discordant reports that warrant further study. PMID- 19554401 TI - In vivo and in vitro effects of fluoroquinolones on lipopolysaccharide-induced pro-inflammatory cytokine production. AB - Fluoroquinolones have been reported to affect cytokine production in vitro. We investigated the effects of fluoroquinolones on lipopolysaccharide (LPS)-induced inflammatory cytokine production in vivo and in vitro. LPS was administered to mice treated with ciprofloxacin, gatifloxacin, norfloxacin, and levofloxacin, and the serum levels of tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), and interleukin 6 (IL-6) were measured. In addition, peritoneal macrophages collected from mice were treated with the four fluoroquinolones for 1 h, followed by the addition of LPS, and the TNF-alpha, IL-1beta, and IL-6 levels in culture fluid were measured. In LPS-treated mice, ciprofloxacin, gatifloxacin, and norfloxacin (100 mg/kg) significantly reduced the serum TNF-alpha level (6.8% 63.6% of control). Levofloxacin at 100 mg/kg did not affect the TNF-alpha level, whereas levofloxacin at a lower dose (10 mg/kg) significantly increased the level. All four fluoroquinolones (100 mg/kg) investigated in this study tended to decrease the serum IL-1beta levels (65.5%-65.9% of control), but this was not a significant change. The serum IL-6 levels were increased in ciprofloxacin administered mice, whereas the other fluoroquinolones did not affect the serum IL 6 levels. In mouse peritoneal macrophages, LPS induced TNF-alpha, IL-1beta, and IL-6 production. Ciprofloxacin, gatifloxacin, and norfloxacin (100 mug/ml) inhibited both TNF-alpha (12.1%-69.0% of control) and IL-1beta production (22.1% 68.8% of control). Levofloxacin (100 mug/ml) inhibited IL-1beta production (65.0% of control), but not TNF-alpha production. LPSstimulated IL-6 production was inhibited only by norfloxacin (59.5 % of control). Our in vivo and in vitro results suggest that fluoroquinolones, especially ciprofloxacin, gatifloxacin, and norfloxacin, which have a cyclopropyl group at the N1 position and/or a piperazinyl group at the C7 position, modify inflammatory responses. PMID- 19554402 TI - Evaluation of the risk factors for febrile neutropenia associated with hematological malignancy. AB - Febrile neutropenia (FN) can frequently become a very serious problem. In 2002, Klastersky and colleagues established the Multinational Association for Supportive Care in Cancer (MASCC) score, which consisted of risk factors for conditions that included solid tumors. However, hematopoietic tumors, in comparison to solid tumors, are plagued by such problems as the quantity and quality of abnormalities associated with leukocytes and neutrophils and the requirement for higher dosages of both radio- and chemotherapy. FN is a complication associated with hematological malignancies that can lead to a fatal outcome, but it is avoidable if the appropriate preventive treatment is performed at an early stage. The subjects of the present study consisted of 354 patients with hematopoietic malignancies who were treated at the Japanese Red Cross Medical Center Hospital, Tokyo, between August 2000 and September 2004. They were retrospectively evaluated for the risk factors of FN by applying Wilcoxon's rank sum test. A scoring index was defined and the patients were classified into high- and low-risk groups before evaluation. The following nine risk factors, which may significantly influence the relationship between the time required for defervescence and the duration of neutropenia - age; hematological diseases; the leukocyte count during the febrile period; the reduction in leukocyte count per day before the onset of FN; the prophylactic administration of antimycotic agents; sterilization of the intestinal tract; and urine albumin content, creatine level, and C-reactive protein (CRP) level - were expressed in points and their sum was termed risk points. The range of risk points was classified as 0-3 and 4-9. The time required for defervescence was 5.1 days when the risk points were in the range of 0-3 and 8.1 days when the points were in the range of 4-9. These figures were distributed normally and there was a significant difference between the two groups (P = 0.0016). FN associated with hematological malignancies is somewhat different from that related to other malignancies; it is therefore associated with unique risk factors. Most of the risk factors used in the present study can be evaluated objectively. At the onset of FN, they were expressed in points for evaluation. Further prospective studies are needed to determine whether these risk factors are suitable for use in actual cases. PMID- 19554400 TI - Nationwide surveillance of bacterial respiratory pathogens conducted by the Japanese Society of Chemotherapy in 2007: general view of the pathogens' antibacterial susceptibility. AB - For the purpose of a nationwide surveillance of the antimicrobial susceptibility of bacterial respiratory pathogens in patients in Japan, the Japanese Society of Chemotherapy conducted their second year survey, during the period from January to August, 2007. A total of 1178 strains were collected from clinical specimens obtained from adult patients with well-diagnosed respiratory tract infections. Susceptibility testing was evaluable for 1108 strains (226 Staphylococcus aureus, 257 Streptococcus pneumoniae, 6 Streptococcus pyogenes, 206 Haemophilus influenzae, 120 Moraxella catarrhalis, 122 Klebsiella pneumoniae, and 171 Pseudomonas aeruginosa). A total of 44 antibacterial agents, including 26 beta lactams (four penicillins, three penicillins in combination with beta-lactamase inhibitors, four oral cephems, eight parenteral cephems, one monobactam, five carbapenems, and one penem), three aminoglycosides, four macrolides (including ketolide), one lincosamide, one tetracycline, two glycopeptides, six fluoroquinolones, and one oxazolidinone were used for the study. Analysis was conducted at the central reference laboratory according to the method recommended by the Clinical and Laboratory Standards Institute (CLSI). The incidence of methicillinresistant Staphylococcus aureus (MRSA) was high, at 59.7%, and the incidences of penicillin-intermediateresistant and -resistant Streptococcus pneumoniae (PISP and PRSP) were 30.4% and 5.1%, respectively. Among Haemophilus influenzae strains, 19.9% of them were found to be beta-lactamase-non-producing ampicillin (ABPC)-intermediately-resistant (BLNAI), 29.1% to be beta-lactamasenon producing ABPC-resistant (BLNAR), and 6.7% to be beta-lactamase-producing ABPC resistant (BLPAR) strains. Extended-spectrum beta-lactamase-producing Klebsiella pneumoniae was not isolated. Two isolates (1.2%) of Pseudomonas aeruginosa were found to be metallo-beta-lactamase-producing strains, including one (0.6%) suspected multidrug-resistant strain showing resistance to imipenem, amikacin, and ciprofloxacin. These data will be a useful reference for future periodic surveillance studies and for investigations to control resistant infections as well. Continued surveillance is required to prevent the further spread of these antimicrobial resistances. PMID- 19554404 TI - Increase of atypical lymphocytes expressing CD4+/CD45RO+ in an infectious mononucleosis-like syndrome associated with hepatitis A virus infection. AB - Subpopulations of regular and atypical lymphocytes in the peripheral blood of a 24-year-old man with an infectious mononucleosis (IM)-like syndrome associated with hepatitis A virus (HAV) infection were analyzed. The ratio of CD4+ to CD8+ cells was in the normal range (1.19 and 1.23 in the regular and atypical lymphocytes, respectively), with no increase in CD8+ cells. The percentage of CD8+/CD11b- cells was not increased in the atypical lymphocytes. However, CD45RO+ was expressed on 86.3% of CD4+ atypical lymphocytes. The present data suggest that atypical lymphocytes expressing CD4+/CD45RO+ may play the role of helper T cells in the immune system in the development of IM-like syndrome associated with HAV infection. PMID- 19554403 TI - Usefulness of initial blood cultures in patients admitted with pneumonia from an emergency department in Japan. AB - Guidelines recommend obtaining blood cultures for all patients admitted with pneumonia. However, recent American studies have reported the low impact of these cultures on antibiotic therapy. Our aim was to investigate the incidence of bacteremia and change of therapy in admitted pneumonia patients from whom blood cultures were obtained in the emergency department (ED). A retrospective, observational, cohort study was conducted on consecutive patients (age >/=12 years) with pneumonia hospitalized through the ED between January 1 and December 31, 2006, in an urban teaching hospital in Japan. Data were collected on antibiotic sensitivities, empirical antibiotics, and changes of antibiotic management. Blood cultures were classified as positive, negative, or contaminant, based on previously established criteria. Out of 164 consecutive cases, blood cultures were positive in 6 patients (3.7%; 95% confidence interval [CI], 0.8% 6.6%), contaminated in 6 (3.7%), and negative in 152 (92.7%). Of the 6 bacteremic patients, 2 cases were likely to have been caused by concomitant diseases. Blood culture results altered therapy for 4 patients (2.4% of 164; 95% CI, 0.7%-6.1%), of whom 2 patients (1.2%; 95% CI, 0.1%-4.3%) had their coverage narrowed, 1 patient (0.6%; 95% CI, 0.0%-3.4%) had coverage broadened, and 1 patient had altered therapy before the drug sensitivities were reported. Considering cost and workload, the overall total annual cost was 758 631 (107 = 1 $US in June 2008). Blood cultures could identify organisms in only a few patients with pneumonia and rarely altered antibiotic management even in patients with positive cultures. It may not be necessary to obtain blood cultures for patients admitted with pneumonia. PMID- 19554405 TI - Urosepsis caused by Edwardsiella tarda. AB - Edwardsiella tarda is a rare causative agent of human infection, predominantly associated with gastroenteritis. We describe a fatal case of urosepsis caused by E. tarda. The patient's underlying condition of advanced uterine cancer may have contributed to the development of the infection. PMID- 19554407 TI - Rapid analysis of Clostridium difficile strains recovered from hospitalized patients by using the slpA sequence typing system. AB - Clostridium difficile is a nosocomial pathogen that is transmissible between patients via hospital staff and via contaminated environmental surfaces. Recently, a typing system based on the slpA sequence for C. difficile was developed. To elucidate the validity and efficacy of the system in the setting of a local hospital, we carried out typing of C. difficile from patients in our hospital using the system. Twenty-eight stool samples obtained from 17 patients with C. difficile-associated diarrhea were investigated. Twenty-two of the 28 samples were positive for C. difficile by stool culture, and they were able to be classified by slpA sequence typing. The smz-1 and smz-2 strains were revealed to be the predominant types in our hospital, accounting for 73% of all strains. The yok-1, yok-2, t25-1, hr-1, and hj2-2 strains were identified in 1 patient each. The smz-1 strain was identified in all wards except for ward D, while smz-2 was identified in 3 of 4 patients in ward D and was restricted to this ward. Nosocomial infection of smz-1 and smz-2 in our hospital was demonstrated. Distinguishing smz-1 from smz-2 by slpA sequence typing clarified the transmission of C. difficile among patients. In conclusion, slpA sequence typing was useful in the setting of a local hospital and may be a powerful tool for the epidemiological study of C. difficile infection. PMID- 19554406 TI - Emphysematous pyelonephritis successfully treated by early intervention using a renoureteral catheter. AB - A 49-year-old woman with poorly controlled type 2 diabetes mellitus was admitted to hospital complaining of fever, vomiting, and lower abdominal pain. Laboratory investigation revealed leukocytopenia, high blood sugar, and pyuria. Pyelonephritis was then diagnosed. Escherichia coli was isolated from blood and urinary cultures. In spite of antimicrobial therapy, the patient's condition deteriorated. A computed tomography scan of the abdomen on the second day of hospitalization revealed the presence of air in the collecting system of the left kidney. Emphysematous pyelonephritis was diagnosed, and a renoureteral catheter was promptly inserted via the left ureter into the affected pelvis of the left kidney. Imipenem, cefotiam, and levofloxacin were administered during the clinical course. This early intervention and the appropriate antimicrobial therapy were effective in resolving the infection. Urinary tract infections should be carefully managed in patients with poorly controlled diabetes mellitus. PMID- 19554408 TI - Rifampicin use in MRSA infections: will it add to the emergence of multidrug resistant tuberculosis in developing countries? PMID- 19554410 TI - Tuberculosis: don't forget to make a complete diagnosis. PMID- 19554412 TI - Surrounding rim formation and reduction in size after radiofrequency ablation for primary breast cancer. AB - PURPOSE: Radiofrequency ablation (RFA) has recently been used to treat small breast cancer. However, there are no data on the long-term morphological features after the procedure. The present study attempts to clarify the characteristics of and changes in the ablated lesion. MATERIALS AND METHODS: A total of 17 breast cancer patients underwent RFA using a single needle featuring an internally cooled electrode; this was followed by whole-breast irradiation and adjuvant systemic therapy. Magnetic resonance imaging (MRI) using a 1.5-T system was performed before and 1, 3, 6, and 12 months after ablation, and the morphological characteristics and the size of the ablated lesion were evaluated. Mammography was also performed for a comparison with the MRI measurement. RESULTS: MRI displayed no residual enhancement of the tumor after RFA; there was an altered signal intensity with peripheral enhancement, however, and the area decreased in size gradually at a rate of 3.3% per month. Mammography showed a ring surrounding a roundish area whose size was equal to that seen with MRI. CONCLUSION: Our current series demonstrated the morphological characteristics on breast imaging after RFA plus radiation therapy. The size of the ablated area decreased over time. These findings are valuable for clinical follow-up of breast cancer patients undergoing RFA. PMID- 19554413 TI - Changes in rectal volume and prostate localization due to placement of a rectum emptying tube. AB - PURPOSE: Rectal volume variation has a crucial effect on prostate localization during external beam radiotherapy for prostate cancer. This study investigated the effect of rectal volume reduction by a rectum-emptying tube (RET) on prostate immobilization. MATERIALS AND METHODS: The study group comprised 21 patients who underwent proton beam treatment for prostate cancer. Sigmoid-shaped flexible plastic RETs were used to drain gases from the rectum. Computed tomography (CT) was performed before and after RET placement at the treatment planning stage and at the beginning of treatment. Prostate displacement and changes in rectal volume were measured on the CT images. The feasibility of RET placement was evaluated during and after the procedure. RESULTS: All 21 patients tolerated the procedure. The rectal volume was significantly lower with a RET than without a RET. The differences in rectal volume between the treatment planning stage and the beginning of treatment were significantly lower with a RET than without. RET placement significantly decreased prostate displacement in the anteroposterior and superoinferior directions but not in the left-right direction. CONCLUSION: RET placement reduced both rectal volume and variation in rectal volume. The procedure reduced displacement of the prostate. RET placement thus appears to be an effective technique for immobilizing the prostate. PMID- 19554415 TI - Leiomyoma of the seminal vesicle. AB - We report a case of leiomyoma of the seminal vesicle in a 74-year-old man who presented with left hemilumbago. Computed tomography and magnetic resonance imaging revealed a mass containing coarse calcification and low signal intensity areas on T1- and T2-weighted images. The clinical features of previously reported cases of leiomyoma of the seminal vesicle are presented, including those of the present case. There remains a lack of consensus regarding surgical resection of leiomyoma of the seminal vesicle. PMID- 19554414 TI - Supraclavicular failure after breast-conserving therapy in patients with four or more positive axillary lymph nodes when prophylactic supraclavicular irradiation is omitted. AB - PURPOSE: The incidence of supraclavicular metastasis as the initial failure and the failure patterns in patients with four or more positive axillary lymph nodes (PALNs) after breast-conserving therapy (BCT) without prophylactic supraclavicular irradiation were investigated. MATERIALS AND METHODS: Between 1991 and 2002, a total of 48 women with four or more PALNs underwent BCT without prophylactic supraclavicular irradiation (33 patients with 4-9 PALNs; 15 patients with > or =10 PALNs). RESULTS: The median follow-up time was 50 months. Among the patients with 4-9 PALNs, 3% had isolated supraclavicular metastasis as the initial failure, and 30% had distant metastasis as the initial failure. Among patients with > or =10 PALNs, 7% had isolated supraclavicular metastasis as the initial failure, and 40% had distant metastasis as the initial failure. The 4 year isolated supraclavicular failure rates were 5% for all patients, 3% for patients with 4-9 PALNs, and 8% for patients with >/=10 PALNs. CONCLUSION: In patients who had undergone BCT and had had four or more PALNs, the major failure pattern was distant failure with or without locoregional failure; isolated supraclavicular failure as the initial failure comprised a less common failure pattern. Omission of prophylactic supraclavicular irradiation may be acceptable for this subset of patients. PMID- 19554416 TI - Primary malignant lymphoma of the breast. AB - Primary malignant lymphoma of the breast is a rare tumor. Recently, we encountered a case of primary malignant lymphoma of the breast and present here the mammographic and ultrasonographic (US) findings with the pathology of this rare breast tumor. The patient was a 47-year-old woman with a left breast mass. Mammography showed a 3.0 x 1.5 cm, lobulated high-density mass with well-defined margins. US revealed a lobulated mass with fairly well-defined borders, hypoechoic and heterogenous internal echoes, and posterior acoustic enhancement. It was difficult to differentiate malignant lymphoma from breast carcinoma by imaging studies. This type of breast cancer should be added to a differential diagnosis when a solitary tumor is found in the breast on mammography. PMID- 19554417 TI - Alveolar hydatid disease of the adrenal gland: computed tomography and magnetic resonance imaging findings. AB - Echinococcosis is a parasitic infection of humans caused by the larval stage of the tapeworm Echinococcus. Primary alveolar echinococcosis of the adrenal gland is rare. We report a case of alveolar hydatid disease of the adrenal gland that presented as a multiloculated cystic mass without calcification. The lesion was purely cystic in nature, suggesting that it was at an early stage of development. PMID- 19554419 TI - Percutaneous management of complications of tuberculous spondylodiscitis: short- to medium-term results. AB - PURPOSE: Psoas abscesses are the most frequent complication of tuberculosis with skeletal involvement. The aim of this paper is to report our experience with the systematic application of percutaneous drainage to tuberculous psoas abscesses. MATERIALS AND METHODS: Between January 1997 and December 2005, 23 patients (14 men and nine women; age range 21-48 years), after a previous study with computed tomography (CT) and/or magnetic resonance (MR) imaging, underwent percutaneous drainage of a tuberculous fluid collection in the psoas muscles. Follow-up consisted of monthly clinical and laboratory assessment, and plain chest radiography and spinal CT every 6-12 months. RESULTS: Spondylodiscitis involved the thoracolumbar spine. Fluid collections were bilateral in 14 cases and communicating in ten of these. Maximum transverse diameter was 7 cm, whereas longitudinal diameter was 14 cm. Placement of the drainage catheter was successful in all cases, and the catheter was left in place for 5-36 (mean 18.4) days. Symptom regression occurred immediately after drainage of the fluid collection. The drainage procedure was curative in 100% of cases. Dislodgement of the drainage catheter occurred in two cases as a result of excessive traction during dressing removal. CONCLUSIONS: A serious complication of bone tuberculosis, psoas abscesses, can be effectively treated by percutaneous drainage, leading to immediate pain resolution. The drainage catheter requires daily monitoring to identify when it can be safely removed without risk of recurrence. PMID- 19554418 TI - Psychiatric comorbidity in patients with chronic daily headache and migraine: a selective overview including personality traits and suicide risk. AB - Studies on the prevalence and impact of psychiatric disorders among headache patients have yielded findings that have clarified the relationship between migraine and major affective disorders, anxiety, illicit drug abuse, nicotine dependence, and suicide attempts. Studies in both clinical and community-based settings have demonstrated an association between migraine and a number of specific psychiatric disorders. In large-scale population-based studies, persons with migraine are from 2.2 to 4.0 times more likely to have depression. In longitudinal studies, the evidence supports a bidirectional relationship between migraine and depression, with each disorder increasing the risk of the other disorder. Although a strong association has been demonstrated consistently for migraine and major depression, especially for migraine with aura, there has been less systematic research on the links between migraine and bipolar disorder. This review will focus on the way in which psychiatric disorders decrease the quality of life and result in a worse prognosis, chronicity of the disease, and a worse response to treatment. Short-term pharmaceutical care intervention improves the patients' mental health, but it does not significantly change the number and severity of headaches. The increase in self-efficacy and mental health associated with pharmaceutical care may be instrumental in improving the long-term pharmacotherapy of patients with migraine and headache. PMID- 19554420 TI - Future directions in percutaneous vertebroplasty. AB - The first percutaneous vertebroplasty, used to treat a painful cervical haemangioma, was performed by a French team in 1984 and reported in the literature in 1987. This technique has rapidly become the standard of care for treatment of medically refractory painful vertebral compression fractures. Vertebral fractures usually become evident because of pain of varying intensity that reduces the patient's quality of life, producing functional limitations, depression, disability, height loss, spinal instability and kyphotic deformity associated with impaired lung capacity. Many diseases may underlie vertebral compression fractures, such as osteoporosis, trauma, neoplasms and haemangioma. Vertebroplasty, as derived from our experience and a review of the literature data, has more than 70%-90% effectiveness for short-term pain reduction and return to activity. The aim of this paper was to describe the state of the art of this spinal interventional radiology procedure and to examine the future directions of percutaneous vertebroplasty. PMID- 19554421 TI - Relationship between 24-h air pollution, emergency department admission and diagnosis of acute coronary syndrome. AB - Current available evidence suggests that air pollution, especially particulate matter <10 mum in aerodynamic diameter (PM10), has acute and chronic effects on health. We aimed to evaluate the epidemiological association between 24-h PM10 levels and acute coronary syndrome (ACS). We analyzed the number of daily admissions at the emergency unit (ED) of patients with suspected ACS, and the number of daily diagnosis of ACS in the same local ED according to the 24-h PM10, over a nearly 3-month period. 209 out of the 1,688 patients admitted to the ED with a clinical suspicion of ACS had a definitive diagnosis of ACS. Whereas the mean number of ED admissions for suspected ACS did not differ significantly according to the 24-h levels of PM10 (18 +/- 1 vs. 17 +/- 1; P = 0.13), the mean number of diagnoses of ACS was 26% higher when the 24-h levels of PM10 were >50 microg/m(3) (2.4 +/- 0.2 vs. 1.9 +/- 0.2; P = 0.03). A statistically significant association was also observed between 24-h PM10 levels and daily number of ACS diagnoses (r = 0.26; P < 0.001), but not between 24-h PM10 levels and daily ED accesses for suspected ACS (r = 0.12; P = 0.24). Taken together, these results support the existence of an epidemiologic association between air pollution and acute cardiovascular events. PMID- 19554423 TI - Involvement of the ornithine decarboxylase/polyamine system in precondition induced cardioprotection through an interaction with PKC in rat hearts. AB - Polyamines (putrescine, spermidine, and spermine) play an essential role in cell growth, differentiation, and apoptosis. Protein kinase C (PKC) stimulates polyamine biosynthesis through the induction of ornithine decarboxylase (ODC), a rate-limiting enzyme in polyamine biosynthesis. Activation of PKC mediates ischemic preconditioning to reduce necrosis and apoptosis in intact hearts and in isolated culture cardiomyocytes. In this study, we examined whether the ODC/polyamine system is involved in the ischemic preconditioning signaling pathway and whether this system interacts with PKC in preconditioning-induced cardioprotection. Hearts were preconditioned with three cycles of 5-min ischemia and 5-min reflow, which caused an increase of ODC expression and spermidine, spermine, and total polyamine pool levels. alpha-Difluoromethylornithine (DFMO) and ethylglyoxal bis (guanylhydrazone) (EGBG) inhibited the key enzymes involved in polyamine biosynthesis, and abolished the preconditioning-induced reduction in infarct size and improvement in postischemic heart contractility function. They also increased cell apoptosis extent and aggravated myocardium ultrastructure damage. Inhibition also attenuated the preconditioning-induced translocation and activation of the PKC-delta, -epsilon isoforms from the cytosol to the particulate. Conversely, activation of PKC by phorbol 12-myristate 13-acetate (PMA) upregulated the ODC/polyamine system, whereas the PKC inhibitor chelerythrine (Che) downregulated the ODC/polyamine system. These findings suggest that upregulation of the polyamine synthesis metabolism occurs in response to preconditioning and mediates preconditioning-induced cardioprotection. The ODC/polyamine system and PKC signals may "cross-talk" in preconditioned hearts such that inhibiting one pathway leads to a reduction in the activity of the other pathway and vice versa. PMID- 19554422 TI - Proteomic studies of rat tibialis anterior muscle during postnatal growth and development. AB - In this study, a proteomic analysis consisting of two-dimensional gel electrophoresis and MALDI-TOF/TOF mass spectrometry was accomplished to investigate the complex protein expression patterns in rat tibialis anterior muscle during postnatal 3-month period. We determined the time-dependent expression alterations of 107 protein spots, among which 53 protein spots were identified. These identified proteins included skeletal contractile proteins, metabolic enzymes, chaperone, intermediate filament, and signal transduction proteins. The time-dependent expression of three proteins, such as Mylpf, desmin, and RKIP, was confirmed by Western blot analysis and immunohistochemistry. The functional implication of these expression changes was also discussed. We further analyzed the linkage and interactions among the differentially expressed proteins (MAPK1, RKIP, AHSG, etc.). Collectively, the results might add to the understanding of the molecular mechanisms regulating postnatal growth and development of rat tibialis anterior muscle. PMID- 19554424 TI - Effect of in vitro exposure of human serum to 3-butyl-1-phenyl-2 (phenyltelluro)oct-en-1-one on oxidative stress. AB - The objective of this study was to verify the effect of the organochalcogen 3 butyl-1-phenyl-2-(phenyltelluro)oct-en-1-one on some parameters of oxidative stress in human serum. Serum of volunteers were incubated for 30 min in the presence or absence of 1, 10, or 30 microM of 3-butyl-1-phenyl-2 (phenyltelluro)oct-en-1-one and oxidative stress was measured. First, we tested the influence of the compound on 1,1-diphenyl-2-picrylhydrazyl (DPPH(*)) radical scavenging and verified that the organotellurium did not have any antioxidant properties. The organochalcogen was capable to enhance TBARS but the compound was not able to alter carbonyl assay. Furthermore, the organochalcogen provoked a reduction of protein thiol groups measured by the sulfhydryl assay. Moreover, the organotellurium enhanced the activity of catalase and superoxide dismutase, inhibited the activity of glutathione peroxidase and did not modify the glutathione S-transferase activity. Furthermore, nitric oxide production and hydroxyl radical activity were not affected by the compound. Our findings showed that this organochalcogen induces oxidative stress in human serum, indicating that this compound is potentially toxic to human beings. PMID- 19554426 TI - Conservation across species identifies several transcriptional enhancers in the HEX genomic region. AB - The HEX gene encodes for a homeodomain-containing transcription factor that controls various phases of vertebrate development. During development, as well as in adult, HEX is expressed in several different tissues including thyroid, liver, lung, mammary gland, haematopoietic progenitors, and endothelial cells, suggesting that this gene is subjected to a complex transcriptional regulation. In this study, we have evaluated the presence of different enhancers in the HEX gene region by using a phylogenetic approach. Several non-coding sequences, conserved between human and mouse, were selected. Four conserved sequences showed enhancer activity in MCF-7 cells. Two of these enhancers (located in the first and third intron, respectively) have been previously identified by other experimental approaches. These elements, as well as one among the new identified enhancers (located 2 kb 3' to the HEX gene), are able to activate the HEX minimal promoter "in trans." The activity of the 3' enhancer was strongly reduced by overexpression of HDAC3. PMID- 19554425 TI - Protective effect of salidroside against H2O2-induced cell apoptosis in primary culture of rat hippocampal neurons. AB - Salidroside, a phenylpropanoid glycoside separated from a medicinal plant Rhodiola rosea, has been documented to have protective effects on neuronal cells in vitro. This study investigated whether salidroside was able to extend its unique neuroprotection to primary cultured rat hippocampal neurons against hydrogen peroxide (H(2)O(2))-induced cell damage. Cell viability tests and cell apoptosis assays confirmed that salidroside pretreatment attenuated H(2)O(2) stimulated apoptotic cell death in primary culture of hippocampal neurons in a concentration-dependent manner. The measurements of caspase-3 activity, nitric oxide (NO) production, and NO synthase (NOS) activity suggest that the protection of salidroside, shown in this study, might be mediated by inhibiting caspase-3 activity, and antagonizing NO production and NOS activity during H(2)O(2) stimulation. Perhaps, this study might contribute to the development of salidroside as a broad-spectrum agent for preventing and/or treating neuronal damage in neurodegenerative disorders. PMID- 19554427 TI - Effect of the oxygen content in solution on the static and cyclic deformation of titanium foams. AB - It is well known that interstitials affect the mechanical properties of titanium and titanium alloys. Their effects on the fatigue properties of titanium foams have not, however, been documented in the literature. This paper presents the effect of the oxygen content on the static and dynamic compression properties of titanium foams. Increasing the oxygen content from 0.24 to 0.51 wt% O in solution significantly increases the yield strength and reduces the ductility of the foams. However, the fatigue limit is not significantly affected by the oxygen content and falls within the 92 MPa +/- 12 MPa range for all specimens investigated in this study. During cyclic loading, deformation is initially coming from cumulative creep followed by the formation of microcracks. The coalescence of these microcracks is responsible for the rupture of the specimens. Fracture surfaces of the specimens having lower oxygen content show a more ductile aspect than the specimens having higher oxygen content. PMID- 19554428 TI - Characterization of pHEMA-based hydrogels that exhibit light-induced bactericidal effect via release of NO. AB - A light-activated NO donor, [Mn(PaPy(3))(NO)]ClO(4) (1a), has been incorporated into HEMA-based polymer hydrogel and the nitrosyl-polymer conjugate materials 1a(x) . HG and 1a(x) . HG(MB) have been characterized. The NO releasing properties and antibacterial capabilities of these materials in conjunction with growth attenuators such as hydrogen peroxide and methylene blue (MB) are reported. Since the nitrosyl releases NO only upon exposure to light, materials like 1a(x) . HG(MB) could be used as wound dressings that deliver NO under controlled conditions. PMID- 19554429 TI - Lactoferrin conjugated with 40-kDa branched poly(ethylene glycol) has an improved circulating half-life. AB - PURPOSE: We developed a lactoferrin conjugate by modifying bovine lactoferrin (bLF) with a 40-kDa branched poly(ethylene glycol) (PEG) molecule (designated 40 k-PEG-bLf), and we evaluated its in vitro activities and pharmacokinetic properties. MATERIALS AND METHODS: We prepared 40k-PEG-bLf by amino conjugation with N-hydroxysuccinimide-activated PEG. This conjugate was purified by cation exchange chromatography and its in vitro biological activities, such as iron binding, anti-inflammatory effects, and resistance to proteolytic enzymes were investigated. In vivo pharmacokinetics analyses, were also performed to examine the rate of clearance from the plasma in rats. RESULTS: The 40k-PEG-bLf conjugate was fully active in iron binding and exhibited 97.1 +/- 5.5% (mean +/- S.E., n = 6) of the original anti-inflammatory activity. The in vitro peptic susceptibility of 40 k-PEG-bLf revealed that the proteolytic half-life increased at least 6-fold that of unmodified LF. This PEGylated conjugate demonstrated a plasma half-life that was 8.7-fold longer than that of the unmodified bLF in rats. CONCLUSIONS: The 40k-PEG-bLf exhibited improved in vitro bioactivity and stability and enhanced pharmacokinetic properties as compared to those of the unmodified bLF and the 20 k-PEG-bLf conjugate, which was recently developed by PEGylation of bLF with a 20-kDa branched PEG [Nojima Y. et al. Bioconjugate Chem. 19:2253-2259 (2008)]. PMID- 19554430 TI - Drug release kinetics and transport mechanisms from semi-interpenetrating networks of gelatin and poly(ethylene glycol) diacrylate. AB - PURPOSE: To elucidate the key parameters affecting solute transport from semi interpenetrating networks (sIPNs) comprised of poly(ethylene glycol) diacrylate (PEGdA) and gelatin that are partially crosslinked, water-swellable and biodegradable. Effects of material compositions, solute size, solubility, and loading density have been investigated. MATERIALS AND METHODS: sIPNs of following gelatin/PEGdA weight-to-weight ratios were prepared: 10:15, 10:20, 10:30, 15:15, 20:15. Five model solutes of different physicochemical properties were selected, i.e. silver sulfadiazine (AgSD), bupivacaine hydrochloride (Bup), sulfadiazine sodium (NaSD), keratinocyte growth factor (KGF), and bovine serum albumin conjugated with fluorescein isothiocyanate (BSA-FITC). Release studies were performed and the results were analyzed using three hydrogel based common theories (free volume, hydrodynamic and obstruction). RESULTS: The release kinetics of model solutes was influenced by each factor under investigation. Specifically, the initial release rates and intra-gel diffusivity decreased with increasing PEGdA content or increasing solute molecular weight. However, the initial release rate and intra-gel diffusivity increased with increasing gelatin content or increasing solute water solubility, which contradicted with the classical hydrogel based solute transport theories, i.e. increasing polymer volume leads to decreased solute diffusivity within the gel. CONCLUSION: This analysis provides structure-functional information of the sIPN as a potential therapeutic delivery matrix. PMID- 19554431 TI - Optimization of the intravenous glucose tolerance test in T2DM patients using optimal experimental design. AB - Intravenous glucose tolerance test (IVGTT) provocations are informative, but complex and laborious, for studying the glucose-insulin system. The objective of this study was to evaluate, through optimal design methodology, the possibilities of more informative and/or less laborious study design of the insulin modified IVGTT in type 2 diabetic patients. A previously developed model for glucose and insulin regulation was implemented in the optimal design software PopED 2.0. The following aspects of the study design of the insulin modified IVGTT were evaluated; (1) glucose dose, (2) insulin infusion, (3) combination of (1) and (2), (4) sampling times, (5) exclusion of labeled glucose. Constraints were incorporated to avoid prolonged hyper- and/or hypoglycemia and a reduced design was used to decrease run times. Design efficiency was calculated as a measure of the improvement with an optimal design compared to the basic design. The results showed that the design of the insulin modified IVGTT could be substantially improved by the use of an optimized design compared to the standard design and that it was possible to use a reduced number of samples. Optimization of sample times gave the largest improvement followed by insulin dose. The results further showed that it was possible to reduce the total sample time with only a minor loss in efficiency. Simulations confirmed the predictions from PopED. The predicted uncertainty of parameter estimates (CV) was low in all tested cases, despite the reduction in the number of samples/subject. The best design had a predicted average CV of parameter estimates of 19.5%. We conclude that improvement can be made to the design of the insulin modified IVGTT and that the most important design factor was the placement of sample times followed by the use of an optimal insulin dose. This paper illustrates how complex provocation experiments can be improved by sequential modeling and optimal design. PMID- 19554432 TI - Nonlinear pharmacokinetics of therapeutic proteins resulting from receptor mediated endocytosis. AB - Receptor mediated endocytosis (RME) plays a major role in the disposition of therapeutic protein drugs in the body. It is suspected to be a major source of nonlinear pharmacokinetic behavior observed in clinical pharmacokinetic data. So far, mostly empirical or semi-mechanistic approaches have been used to represent RME. A thorough understanding of the impact of the properties of the drug and of the receptor system on the resulting nonlinear disposition is still missing, as is how to best represent RME in pharmacokinetic models. In this article, we present a detailed mechanistic model of RME that explicitly takes into account receptor binding and trafficking inside the cell and that is used to derive reduced models of RME which retain a mechanistic interpretation. We find that RME can be described by an extended Michaelis-Menten model that accounts for both the distribution and the elimination aspect of RME. If the amount of drug in the receptor system is negligible a standard Michaelis-Menten model is capable of describing the elimination by RME. Notably, a receptor system can efficiently eliminate drug from the extracellular space even if the total number of receptors is small. We find that drug elimination by RME can result in substantial nonlinear pharmacokinetics. The extent of nonlinearity is higher for drug/receptor systems with higher receptor availability at the membrane, or faster internalization and degradation of extracellular drug. Our approach is exemplified for the epidermal growth factor receptor system. PMID- 19554433 TI - A probabilistic approach for the evaluation of pharmacological effect induced by patient irregular drug intake. AB - Fine individual drug intake data, generally collected by electronic monitoring devices, reveal that individual marked random patterns are likely to persist through long therapeutic periods. This work aims to establish the relationship between irregularity in drug intake and its potential impact on therapeutic outcomes, which will also serve as a basis for more objective interventions. First we proposed a direct way to extract the necessary information representing the patient drug intake history. To provide a fair evaluation of the pharmacological performance, we revisited several classical pharmacological indices and proposed new ones in the stochastic context of patient's drug intake irregularity. To illustrate our procedure, we have considered two cases of HIV treatment using a combination of lopinavir/ritonavir (Kaletra@) for once daily and twice daily regimens. We have quantified the impact on therapeutic effect of various characteristics in dosing histories, namely missing doses and deviations from nominal times. Using our newly defined pharmacological indices, we clearly showed the ability of our probabilistic approach in measuring the impact of noncompliance. As a direct fallout, we have discussed strategies to attenuate the impact of noncompliance through an optimal design of dosing regimen. PMID- 19554434 TI - Effect of pendant group length upon metal ion complexation in acetonitrile by di ionized calix[4]arenes bearing two dansyl fluorophores. AB - A series of three di-ionizable calix[4]arenes with two pendant dansyl (1 dimethylaminonaphthalene-5-sulfonyl) groups linked to the lower rims was synthesized. Structures of the three ligands were identical except for the length of the spacers which connected the two dansyl groups to the calix[4]arene scaffold. Following conversion of the ligands into their di-ionized di(tetramethylammonium) salts, absorption and emission spectrophotometry were utilized to probe the influence of metal cation (Li+, Na+, K+, Rb+, Cs+, Mg2+, Ca2+, Sr2+, Ba2+, Ag+, Cd2+, Co2+, Fe2+, Hg2+, Mn2+, Pb2+, Zn2+ and Fe3+) complexation in acetonitrile. Upon complexation with these metal cations, emission spectra underwent marked red shifts and quenching of the dansyl group fluorescence for the di-ionized ligand with the shortest spacer. A similar effect was noted for the di-ionized ligand with an intermediate spacer for all of the metal ions, except Ba2+. For the di-ionized ligand with the longest spacer, the metal cations showed different effects on the emission spectrum. Li+, Mg2+, Ca2+ and Ba2+ caused enhancement of emission intensity with a red shift. Other metal cations produce quenching with red shifts in the emission spectra. Transition metal cations interacted strongly with all three di-ionized ligands. In particular, Fe3+ and Hg2+ caused greater than 99% quenching of the dansyl fluorescence in the di-ionized ligands. PMID- 19554435 TI - County-level social environment determinants of health-related quality of life among US adults: a multilevel analysis. AB - To show that an individual's health-related quality of life (HRQOL) is not determined only by their personal-level characteristics, but also is socially determined by both physical and social environmental characteristics of their communities. This analysis examined the association of selected county-level indicators on respondents' unhealthy days and assessed the utility of mean unhealthy days for US counties as community health indicators. Data came from the 1999-2001 Behavioral Risk Factor Surveillance System. We used multilevel models to calculate the proportion of between-county variation in HRQOL that was explained by county-level contextual variables and examine the causal heterogeneity of some personal-level factors modified by these contextual variables. Counties with worse socioeconomic indicators, high mortality rate, and low life expectancy were associated with higher numbers of unhealthy days. These indicators explained 13-22% variance of county-level physically unhealthy days and 4.5-9.5% variance of county-level mentally unhealthy days. The GINI index, suicide rate, percent uninsured, primary care facilities-to-population ratio, and most county-level demographic and housing indicators also had significant but smaller impact on respondents' unhealthy days. Also, the counties with poorer socioeconomic scores had additional negative HRQOL impact on older persons. This study provides important new empirical information on whether various commonly measured characteristics of the social environment, which are believed to be social determinants of health, are in fact associated with the perceived physical and mental health of its residents. Our findings provide additional support for the construct validity of county-level HRQOL as a community health indicator. PMID- 19554436 TI - A qualitative study of women's experiences of communication in antenatal care: identifying areas for action. AB - To identify key features of communication across antenatal (prenatal) care that are evaluated positively or negatively by service users. Focus groups and semi structured interviews were used to explore communication experiences of thirty pregnant women from diverse social and ethnic backgrounds affiliated to a large London hospital. Data were analysed using thematic analysis. Women reported a wide diversity of experiences. From the users' perspective, constructive communication on the part of health care providers was characterised by an empathic conversational style, openness to questions, allowing sufficient time to talk through any concerns, and pro-active contact by providers (e.g. text message appointment reminders). These features created reassurance, facilitated information exchange, improved appointment attendance and fostered tolerance in stressful situations. Salient features of poor communication were a lack of information provision, especially about the overall arrangement and the purpose of antenatal care, insufficient discussion about possible problems with the pregnancy and discourteous styles of interaction. Poor communication led some women to become assertive to address their needs; others became reluctant to actively engage with providers. General Practitioners need to be better integrated into antenatal care, more information should be provided about the pattern and purpose of the care women receive during pregnancy, and new technologies should be used to facilitate interactions between women and their healthcare providers. Providers require communications training to encourage empathic interactions that promote constructive provider-user relationships and encourage women to engage effectively and access the care they need. PMID- 19554437 TI - Children with special health care needs: how immigrant status is related to health care access, health care utilization, and health status. AB - To compare health care access, utilization, and perceived health status for children with SHCN in immigrant and nonimmigrant families. This cross-sectional study used data from the 2003 California Health Interview Survey to identify 1404 children (ages 0-11) with a special health care need. Chi-square and logistic regression analyses were used to examine relations between immigrant status and health access, utilization, and health status variables. Compared to children with special health care needs (CSHCN) in nonimmigrant families, CSHCN in immigrant families are more likely to be uninsured (10.4 vs. 4.8%), lack a usual source of care (5.9 vs. 1.9%), report a delay in medical care (13.0 vs. 8.1%), and report no visit to the doctor in the past year (6.8 vs. 2.6%). They are less likely to report an emergency room visit in the past year (30.0 vs. 44.0%), yet more likely to report fair or poor perceived health status (33.0 vs. 16.0%). Multivariate analyses suggested that the bivariate findings for children with SHCN in immigrant families largely reflected differences in family socioeconomic status, parent's language, parental education, ethnicity, and children's insurance status. Limited resources, non-English language, and limited health care use are some of the barriers to staying healthy for CSHCN in immigrant families. Public policies that improve access to existing insurance programs and provide culturally and linguistically appropriate care will likely decrease health and health care disparities for this population. PMID- 19554438 TI - Hurricane Katrina-related maternal stress, maternal mental health, and early infant temperament. AB - To investigate temperament in infants whose mothers were exposed to Hurricane Katrina and its aftermath, and to determine if high hurricane exposure is associated with difficult infant temperament. A prospective cohort study of women giving birth in New Orleans and Baton Rouge, LA (n = 288) in 2006-2007 was conducted. Questionnaires and interviews assessed the mother's experiences during the hurricane, living conditions, and psychological symptoms, 2 months and 12 months postpartum. Infant temperament characteristics were reported by the mother using the activity, adaptability, approach, intensity, and mood scales of the Early Infant and Toddler Temperament Questionnaires, and "difficult temperament" was defined as scoring in the top quartile for three or more of the scales. Logistic regression was used to examine the association between hurricane experience, mental health, and infant temperament. Serious experiences of the hurricane did not strongly increase the risk of difficult infant temperament (association with three or more serious experiences of the hurricane: adjusted odds ratio (aOR) 1.50, 95% confidence interval (CI) 0.63-3.58 at 2 months; 0.58, 0.15-2.28 at 12 months). Maternal mental health was associated with report of difficult infant temperament, with women more likely to report having a difficult infant temperament at 1 year if they had screened positive for PTSD (aOR 1.82, 95% confidence interval (CI) 0.61-5.41), depression, (aOR 3.16, 95% CI 1.22-8.20) or hostility (aOR 2.17, 95% CI 0.81-5.82) at 2 months. Large associations between maternal stress due to a natural disaster and infant temperament were not seen, but maternal mental health was associated with reporting difficult temperament. Further research is needed to determine the effects of maternal exposure to disasters on child temperament, but in order to help babies born in the aftermath of disaster, the focus may need to be on the mother's mental health. PMID- 19554439 TI - The UNC-CH MCH Leadership Training Consortium: building the capacity to develop interdisciplinary MCH leaders. AB - This article describes the UNC-CH MCH Leadership Consortium, a collaboration among five MCHB-funded training programs, and delineates the evolution of the leadership curriculum developed by the Consortium to cultivate interdisciplinary MCH leaders. In response to a suggestion by the MCHB, five MCHB-funded training programs--nutrition, pediatric dentistry, social work, LEND, and public health- created a consortium with four goals shared by these diverse MCH disciplines: (1) train MCH professionals for field leadership; (2) address the special health and social needs of women, infants, children and adolescents, with emphasis on a public health population-based approach; (3) foster interdisciplinary practice; and (4) assure competencies, such as family-centered and culturally competent practice, needed to serve effectively the MCH population. The consortium meets monthly. Its primary task to date has been to create a leadership curriculum for 20-30 master's, doctoral, and post-doctoral trainees to understand how to leverage personal leadership styles to make groups more effective, develop conflict/facilitation skills, and identify and enhance family-centered and culturally competent organizations. What began as an effort merely to understand shared interests around leadership development has evolved into an elaborate curriculum to address many MCH leadership competencies. The collaboration has also stimulated creative interdisciplinary research and practice opportunities for MCH trainees and faculty. MCHB-funded training programs should make a commitment to collaborate around developing leadership competencies that are shared across disciplines in order to enhance interdisciplinary leadership. PMID- 19554440 TI - The health and cognitive growth of Latino toddlers: at risk or immigrant paradox? AB - Epidemiologists have shown how birth outcomes are generally robust for immigrant Latina mothers, despite often situated in poor households, advanced by their strong prenatal and nutritional practices. But little is known about (1) how these protective factors may differ among Latino subgroups, (2) the extent to which birth outcomes, ongoing maternal practices, and family supports advance Latino toddlers' health and physical growth, and (3) whether the same processes advance toddlers' early cognitive growth. We drew on a national probability sample of 8,114 infants born in 2001, including 1,450 of diverse Latino origins. Data come from birth records, maternal interviews when the child was 9 and 24 months of age, and direct assessments of health status, physical growth, and cognitive proficiencies. Descriptive analyses compared Mexican-heritage and other Latino mothers and toddlers relative to middle-class whites. Multivariate regression techniques identified predictors of child health, weight, and BMI, as well as cognitive proficiencies at 24 months. Infants of Mexican-heritage or less acculturated Latina mothers displayed robust birth outcomes, compared with other ethnic groups. The low incidence of premature births and low birthweight among these mothers continued to advance their cognitive growth through 24 months of age. Yet Latino children overall displayed smaller gains in cognitive proficiencies between 9 and 24 months, compared with middle-class populations, attributable to Latinas' lower levels of maternal education, weaker preliteracy practices, and a higher ratio of children per resident adult. Health practitioners should recognize that many Latina mothers display healthy prenatal practices and give birth to robust infants. But these early protective factors do not necessarily advance early cognitive growth. Screening practices, early interventions, and federal policy should become more sensitive to these countervailing dynamics. PMID- 19554441 TI - Hyponatremia in heart failure: the role of arginine vasopressin and diuretics. AB - PURPOSE: To provide an overview of the role of arginine vasopressin (AVP) in the development of hyponatremia in patients with heart failure (HF), the role of diuretics, and the potential for vasopressin-receptor antagonists in the treatment of HF. METHODS: A MEDLINE literature search was performed to identify articles relating to HF, diuretics, hyponatremia, AVP, and vasopressin-receptor antagonists. DISCUSSION: HF affects more than 5 million patients in the United States and is associated with substantial cost, morbidity, and mortality. One of the complications associated with HF, as well as with its treatment, is the development of hyponatremia. Hyponatremia in patients with HF is associated with poor outcomes and can limit the use of diuretic therapy. AVP is the primary stimulus to the development of hyponatremia in these patients and therapies that target AVP action would seem a logical choice in the therapeutic regimen for HF. Drugs that antagonize the vasopressin V(2) receptor, which is primarily responsible for water resorption in the kidney, are now available and have been studied in patients with HF. These drugs have been associated with improvements in serum sodium concentration, urine output, and body weight, but have shown no long-term mortality benefit in patients with HF. In a subset of patients with baseline hyponatremia, these agents improved quality of life scores. CONCLUSION: Vasopressin-receptor antagonists may prove useful in the treatment of HF; however, the exact role of these agents in the treatment of HF still requires further study. PMID- 19554442 TI - Efficient transduction of cytotoxic and anti-HIV-1 genes by a gene-regulatable lentiviral vector. AB - Lentiviral vectors modified from human immunodeficiency virus type 1 (HIV-1) offer a promising approach for gene therapy, facilitating transduction of genes into non-dividing cells both in vitro and in vivo. When transducing cytotoxic or anti-HIV genes, however, the vector must avoid self-inhibition by the transgene that can lead to a disruption in production of infectious virions. In this study, we constructed two HIV-1-based lentiviral vectors harboring the mifepristone inducible gene expression unit in either the forward or the reverse orientation with respect to the direction of viral genomic RNA. The ability of these vectors to transduce cytotoxic and anti-HIV genes was evaluated. When human CD14 was used as a transgene, infectious lentiviral vectors were produced by both forward and reverse vector systems. CD14 expression was efficiently induced in cells transduced by both lentiviral vectors following treatment with mifepristone. However, a higher level of basal transgene expression was observed in the forward vector system in the absence of mifepristone. In contrast, high titers of infectious lentiviral vector containing the cytotoxic vesicular stomatitis virus M gene were successfully generated using the reverse vector, but not the forward vector. In addition, when a VPS4Bdominant negative mutant against HIV-1 budding was cloned into the reverse vector, significant amounts of lentiviral vector were obtained. Subsequent transduction of cells with the VPS4B mutant resulted in approximately 50% inhibition of HIV-1 production only in the presence of mifepristone. Our study thus demonstrates that incorporation of a mifepristone regulatable gene expression unit in the reverse orientation makes significant advances toward development of a lentiviral vector that allows transduction of harmful genes. PMID- 19554443 TI - Promoter motifs essential to the differential transcription of structural and non-structural genes of the white spot syndrome virus. AB - Chimeric reporter genes were generated comprising nine different promoters of the white spot syndrome virus linked to luciferase, with the aim to compare their transcriptional activities in insect cells. The promoters included the four non structural genes DNA polymerase, ribonucleotide reductase small subunit, ribonucleotide reductase large subunit, and thymidine-thymidylate kinase, and the five structural genes VP15, VP19, VP24, VP26, and VP28. The promoters of the non structural but not the structural genes can function in these cells, indicating that transcription of the non-structural genes can be recognized by cellular transcriptional machineries. While the structural genes were highly expressed in natural host cells during white spot syndrome virus infection, their promoters failed to direct transcription in insect cells, suggesting that transcription of these late genes may require virally induced host factor(s). Motifs essential for transcription of the above non-structural genes were identified by transient transfection of insect cells with constructs containing a series of deletions in the 50 terminal region and within the promoter. The minimal promoter sequences of these four genes were also capable of driving expression of the enhanced green fluorescent protein in insect cells. PMID- 19554444 TI - Socio-demographic correlates of fear of crime and the social context of contemporary urban China. AB - Previous research in the West has established major socio-demographic correlates of fear of crime. The interpretation of these correlates is typically based on the concept of physical or social vulnerability of individuals. These correlates are implicitly regarded as invariant to social or community contexts, reflecting universal human behavioral patterns. The present study argues that social change may alter patterns of perceptions associated with fear among socio-demographic groups, thus affecting socio-demographic correlates of fear of crime. We explore how social changes in China have created a generational gap that influences the effects of age and education on fear of crime. The study finds that, in contrast with the well-established patterns in Western communities, the young and educated exhibit a higher level of fear of crime in urban China than their counterparts. The study also finds that consistent with Western literature, females are fearful and that personal victimization experience increases the level of fear. We discuss the social and community processes that produce these interesting patterns. PMID- 19554445 TI - Exposure to violence, support needs, adjustment, and motivators among Guatemalan humanitarian aid workers. AB - Indigenous aid workers carry out the majority of humanitarian aid work, yet there is little empirical information available on their support needs in different contexts. Focus groups (N = 26: Study 1) and a survey (N = 137; Study 2) were conducted with Guatemalan aid workers to explore their exposure to violence, posttraumatic stress symptoms, burnout, support needs, and motivators. Participants reported experiencing an average of 13 events of community violence and 17% reported symptoms consistent with posttraumatic stress disorder (PTSD). Direct community violence exposure and levels of emotional exhaustion were positively related to PTSD symptoms, while levels of personal accomplishment were inversely related to PTSD symptoms. Expressed support needs, motivators and rewards for aid work in the face of adversity are also reported as potential protective factors for further exploration. Implications for training and support of aid workers in similar contexts are also suggested. PMID- 19554446 TI - Eco-toxic effects of sulfadiazine sodium, sulfamonomethoxine sodium and enrofloxacin on wheat, Chinese cabbage and tomato. AB - Investigation of the toxic effects of three veterinary drugs [sulfadiazine sodium (SDS), sulfamonomethoxine sodium (SMMS), and enrofloxacin (EFLX)] on seed germination, root elongation and shoot elongation of wheat (Triticum aestivum L.), Chinese cabbage (Brassica campestris L.) and tomato (Cyphomandra betacea) was carried out. Significant linear relationships between the root and shoot elongation and the concentration of veterinary drugs addition were observed. The effects of the three veterinary drugs on seed germination of wheat, Chinese cabbages and tomato were not significant (P > 0.05), but on shoot and root elongation they were markedly significant (P < 0.05). The inhibitory rates of veterinary drugs on root and shoot elongation of crops were significantly stronger than that on seed germination. Based on IC(50) (drugs concentration when 50% plants show inhibition) of root elongation, wheat was the most sensitive plant to the toxicity of SDS with a IC(50) value as high as 28.1 mg/kg; Chinese cabbage was the most sensitive plant to the toxicity of SMMS with a IC(50) value as high as 27.1 mg/kg; tomato was the most sensitive plant to the toxicity of EFLX with a IC(50) value as high as 125.7 mg/kg. The toxic effects of sulfadiazine sodium and sulfamonometh-oxine sodium on the three crops were much higher than that of enrofloxacin. PMID- 19554447 TI - Immersing practitioners in the recovery model: an educational program evaluation. AB - The ascendance of the recovery movement in mental health care has led to the development and implementation of educational curricula for mental health providers to assist in mental health care system transformation efforts. The Medical College of Georgia (MCG) partnered with the Georgia State Department of Human Resources (DHR) to develop, implement, and evaluate such an educational curriculum for providers within an academic medical institution. This effort, entitled Project GREAT, led to the creation of a curriculum based on the SAMHSA defined (2006) critical components of recovery. As an initial evaluation of educational curriculum effectiveness, the authors examined effects of the training program on recovery-based knowledge and recovery-consistent attitudes. We also compared MCG provider knowledge and attitudes to those of a similar group of providers at a neighboring medical institution who did not receive the intervention and training. Findings generally supported the effectiveness of the intervention in increasing providers' knowledge of recovery and a shift in recovery-supporting attitudes. PMID- 19554448 TI - Eosinophilic esophagitis. AB - Eosinophilic esophagitis is a chronic inflammatory disorder characterized by dense eosinophilic infiltration of the esophageal mucosa. The pathogenesis is incompletely understood and food allergies and aeroallergens have been implicated. The most common clinical presentation in adults is dysphagia to solids. Its associated endoscopic findings are distinct and include concentric rings and longitudinal furrows, although endoscopy may be unremarkable in a minority of patients. A number of management strategies exist; however, data are limited in adults, and only a few are based on randomized controlled trials. Management options include dietary modifications, pharmacological therapy, and endoscopic dilation. PMID- 19554450 TI - Modeling the instantaneous pressure-volume relation of the left ventricle: a comparison of six models. AB - Simulations are useful to study the heart's ability to generate flow and the interaction between contractility and loading conditions. The left ventricular pressure-volume (PV) relation has been shown to be nonlinear, but it is unknown whether a linear model is accurate enough for simulations. Six models were fitted to the PV-data measured in five sheep and the estimated parameters were used to simulate PV-loops. Simulated and measured PV-loops were compared with the Akaike information criterion (AIC) and the Hamming distance, a measure for geometric shape similarity. The compared models were: a time-varying elastance model with fixed volume intercept (LinFix); a time-varying elastance model with varying volume intercept (LinFree); a Langewouter's pressure-dependent elasticity model (Langew); a sigmoidal model (Sigm); a time-varying elastance model with a systolic flow-dependent resistance (Shroff) and a model with a linear systolic and an exponential diastolic relation (Burkh). Overall, the best model is LinFree (lowest AIC), closely followed by Langew. The remaining models rank: Sigm, Shroff, LinFix and Burkh. If only the shape of the PV-loops is important, all models perform nearly identically (Hamming distance between 20 and 23%). For realistic simulation of the instantaneous PV-relation a linear model suffices. PMID- 19554449 TI - A 50-year review of colorectal cancer in African Americans: implications for prevention and treatment. AB - BACKGROUND: African-Americans (AA) have the highest rate of colorectal cancer (CRC) incidence and mortality in the US. CRC in AA is more advanced and right sided. Although screening has been shown to reduce mortality from CRC in the general US population, AA continue to experience a disproportionately higher CRC death compared to other ethnic groups. This study aimed at assessing the trend of CRC in AA, focusing on the changing pattern of in situ tumors in this ethnic group and how observed trends may guide current and future preventive and treatment strategies. MATERIALS AND METHODS: All pathologic reports from 1959 to 2006 in Howard University Hospital (n = 150,000) were reviewed manually. The pathology reports showing colorectal cancer were carefully reviewed and selected by a GI pathologist. Intraepithelial or intramucosal carcinomas were diagnosed as in situ carcinoma. Reviewed pathological information were entered into Microsoft Excel and checked for duplication and missing data. Differences in situ and advanced cancer by sex, histology, location, and years of diagnosis were assessed by Chi-square test. RESULTS: A total of 1,753 CRC cases were diagnosed in this period. About 56% of the cases were female and 51% of the tumors were left-sided. Mean (SD) age was 66 (13) years. The frequency of in situ tumor was 5.8% in this period. There was no statistically significant difference between in situ and advance tumor by age, sex, and tumor location. The rate of in situ tumor peaked in the 1990s at 8.5% (P = 0.0001). We observed a decade-to-decade increasing rate of right-sided tumors, which started at 36% in the period 1959-1970 and peaked in the period of 2001-2006 at 60% (P = 0.0001). CONCLUSIONS: The recent increasing number of advanced and right-sided tumor in our study is concordant with SEER data and has great importance in developing CRC prevention and treatment strategies for AA population. PMID- 19554451 TI - Regulation of Drosophila melanogaster pro-apoptotic gene hid. AB - Key decisions one makes in a lifetime include whether and how often to reproduce, what role to play in the community and, under certain conditions, whether to live or die. Similar decisions are also made at the level of cells: whether to divide, what fate to assume in the multicellular context of metazoan development and, under certain conditions, whether to live or to die. The pro-apoptotic gene hid plays an important role in the execution of cell death in Drosophila. Here, we review the various levels of control that exist to regulate Hid according to the life-or-death choice of a cell. PMID- 19554452 TI - Microfluidic compartmentalized co-culture platform for CNS axon myelination research. AB - This paper presents a circular microfluidic compartmentalized co-culture platform that can be used for central nervous system (CNS) axon myelination research. The microfluidic platform is composed of a soma compartment and an axon/glia compartment connected through arrays of axon-guiding microchannels. Myelin producing glia, oligodendrocytes (OLs), placed in the axon/glia compartment, interact with only axons but not with neuronal somata confined to the soma compartment, reminiscent to in vivo situation where many axon fibres are myelinated by OLs at distance away from neuronal cell bodies. Primary forebrain neurons from embryonic day 16-18 rats were cultured inside the soma compartment for two weeks to allow them to mature and form extensive axon networks. OL progenitors, isolated from postnatal day 1-2 rat brains, were then added to the axon/glia compartment and co-cultured with neurons for an additional two weeks. The microdevice showed fluidic isolation between the two compartments and successfully isolated neuronal cell bodies and dendrites from axons growing through the arrays of axon-guiding microchannels into the axon/glia compartment. The circular co-culture device developed here showed excellent cell loading characteristics where significant numbers of cells were positioned near the axon guiding microchannels. This significantly increased the probability of axons crossing these microchannels as demonstrated by the more than 51 % of the area of the axon/glia compartment covered with axons two weeks after cell seeding. OL progenitors co-cultured with axons inside the axon/glia compartment successfully differentiated into mature OLs. These results indicate that this device can be used as an excellent in vitro co-culture platform for studying localized axon glia interaction and signalling. PMID- 19554453 TI - Designing and implementing an effective pediatric weight management program. AB - In order to successfully address the problem of childhood obesity, effective weight management programs must be established by a variety of clinicians to treat the increasing numbers of overweight and obese children and adolescents. An effective program will: 1) identify children and adolescents with health risks related to excess fat, 2) help families make permanent healthy lifestyle changes, and 3) provide ongoing care to optimize long-term health. Building a pediatric weight management program which achieves these goals requires attention to several aspects, including location, personnel, equipment, institutional support, marketing, referral base, focus of the overall approach, and billing & reimbursement. Maintaining the program after it is established can be enhanced by concentrating on team communication, continuing education, continuous quality improvement, and improving adherence. PMID- 19554454 TI - Exploring the potential of a highly compressible microcrystalline cellulose as novel tabletting excipient in the compaction of extended-release coated pellets containing an extremely water-soluble model drug. AB - Compaction of controlled-release coated pellets into tablets is challenging because of the fusion of pellets and the rupturing of coated film. The difficulty in compaction intensifies with the use of extremely water-soluble drugs. Therefore, the present study was conducted to prepare and compact pellets containing pseudoephedrine hydrochloride as an extremely water-soluble model drug. The pellets were produced using an extrusion-spheronization technique. The drug-loaded pellets were coated to extend the drug release up to 12-h employing various polymers, and then they were compressed into tablets using microcrystalline cellulose Ceolus KG-801 as a novel tabletting excipient. The in vitro drug release studies of coated pellets and tablets were undertaken using the USP basket method in dissolution test apparatus I. The amount of drug released was analyzed at a wavelength of 215 nm. The combined coatings of hydroxypropyl methylcellulose and Kollicoat SR-30D yielded 12-h extended-release pellets with drug release independent of pH of dissolution medium following zero order kinetics. The drug release from the tablets prepared using inert Celous KG 801 granules as tabletting excipient was found faster than that of coated pellets. However, a modification in drug release rate occurred with the incorporation of inert Ceolus KG-801 pellets. The drug dissolution profile from tablets containing 40% w/w each of coated pellets and inert granules along with 20% w/w inert pellets was found to be closely similar to that of coated pellets. Furthermore, the friability, tensile strength, and disintegration time of the tablets were within the USP specifications. PMID- 19554455 TI - Socioeconomic indicators and cardiovascular disease incidence among Japanese community residents: the Jichi Medical School Cohort Study. AB - BACKGROUND: There has been little research in inequalities in risk of cardiovascular disease incidence by social class in Asia. PURPOSE: The purpose of this study was to examine the association between socioeconomic indicators and risk of stroke and coronary heart disease in Japan. METHOD: Data from the Jichi Medical School Study, a population-based prospective cohort study of approximately 11,000 Japanese men and women, were used. The average follow-up period was 11.7 years. Age- and area-adjusted hazard ratios with 95% confidence intervals (CIs) for education level/occupation were calculated using Cox proportional hazard regression analysis. RESULTS: Compared to those who completed education at age 14 or younger, the age and area-adjusted hazard ratios of intraparenchymal hemorrhage incidence for men who completed education at age 15 17 and at age 18 or older were 0.42 (95% CI, 0.21-0.84) and 0.34 (95% CI, 0.14 0.84), respectively. The age- and area-adjusted hazard ratios of intraparenchymal hemorrhage and subarachnoid hemorrhage incidence for female white-collar workers compared to female blue-collar workers were 0.28 (95% CI, 0.08-0.98) and 3.23 (95% CI, 1.29, 8.01), respectively. No associations were found between education level and risk of coronary heart disease among both men and women. CONCLUSION: These results suggest the pattern of social inequalities in health in Japan might be different from that in Western countries. PMID- 19554456 TI - Molecular evolution of a Drosophila homolog of human BRCA2. AB - The human cancer susceptibility gene, BRCA2, functions in double-strand break repair by homologous recombination, and it appears to function via interaction of a repetitive region ("BRC repeats") with RAD-51. A putatively simpler homolog, dmbrca2, was identified in Drosophila melanogaster recently and also affects mitotic and meiotic double-strand break repair. In this study, we examined patterns of repeat variation both within Drosophila pseudoobscura and among available Drosophila genome sequences. We identified extensive variation within and among closely related Drosophila species in BRC repeat number, to the extent that variation within this genus recapitulates the extent of variation found across the entire animal kingdom. We describe patterns of evolution across species by documenting recent repeat expansions (sometimes in tandem arrays) and homogenizations within available genome sequences. Overall, we have documented patterns and modes of evolution in a new model system of a gene which is important to human health. PMID- 19554457 TI - Genotypic variation in element concentrations in brown rice from Yunnan landraces in China. AB - The mineral elements present in brown rice play an important physiological role in global human health. We investigated genotypic variation of eight of these elements (P, K, Ca, Mg, Fe, Zn, Cu, and Mn) in 11 different grades of brown rice on the basis of the number and distance coefficients of 282 alleles for 20 simple sequence repeat (SSR) markers. Six-hundred and twenty-eight landraces from the same field in Yunnan Province, one of the largest centers of genetic diversity of rice (Oryza sativa L.) in the world, formed our core collection. The mean concentrations (mg kg(-1)) of the eight elements in brown rice for these landraces were P (3,480) > K (2,540) > Mg (1,480) > Ca (157) > Zn (32.8) > Fe (32.0) > Cu (13.6) > Mn (13.2). Mean P concentrations in brown rice were 6.56 times total soil P, so the grains are important in tissue storage of P, but total soil K is 7.82 times mean K concentrations in brown rice. The concentrations of the eight elements in some grades of brown rice, on the basis of the number and distance coefficients of alleles for 20 SSR markers for the landraces, were significantly different (P < 0.05), and further understanding of the relationship between mineral elements and gene diversity is needed. There was large variation in element concentrations in brown rice, ranging from 2,160 to 5,500 mg P kg(-1), from 1,130 to 3,830 mg K kg(-1), from 61.8 to 488 mg Ca kg(-1), from 864 to 2,020 mg Mg kg(-1), from 0.40 to 147 mg Fe kg(-1), from 15.1 to 124 mg Zn kg(-1), from 0.10 to 59.1 mg Cu kg(-1), and from 6.7 to 26.6 mg Mn kg(-1). Therefore, germplasm evaluations for Ca, Fe, and Zn concentrations in rice grains have detected up to sevenfold genotypic differences, suggesting that selection for high levels of Ca, Fe, and Zn in breeding for mass production is a feasible approach. Increasing the concentrations of Ca, Fe, and Zn in rice grains will help alleviate chronic Ca, Zn, and Fe deficiencies in many areas of the world. PMID- 19554458 TI - Groundwater vulnerability to selenium in semi-arid environments: Amman Zarqa Basin, Jordan. AB - An evaluation of approximately 250 samples of groundwater in the Amman Zarqa Basin for selenium along with other major and trace elements showed that concentrations of Se ranged between 0.09 and 742 microg/L, with an average value of about 24 microg/L. Selenium concentrations exceeded the recommended threshold for drinking water of the World Health Organization (WHO; 10 micro/L of Se) in 114 samples, with greater than 50 microg/L (quantity equivalent to the Jordanian standard of the allowed concentration of the element in water) of Se in nine cases. The average concentrations of Se in the lower, middle, and upper aquifers of the basin were 3.41, 32.99, and 9.19 microg/L, respectively. Based on the correlation with geologic formations and the statistical analysis of major/minor constituents and Piper tri-linear diagrams, we suggest that carbonate/phosphate dissolution, oxidation-reduction processes, and fertilizers/irrigation return flow are, together, the primary factors affecting the chemistry of the groundwater. Factor analysis helped to define the relative role of limestone dolomitic dissolution in the aquifers (calcium, magnesium, and bicarbonate), agricultural activities (sulfate, nitrates, phosphorus, and potassium), oxidation reduction factor (Eh, Fe, Mn, Cu, Zn, and Se), and anthropogenic (industrial) factor (EC, Fe, Cr, Co, Zn, and As). The high variability in Se concentrations might be related to the possibility of a multi-source origin of Se in the area. PMID- 19554459 TI - Influence of magnetic field on Cr(VI) adsorption capability of given anaerobic sludge. AB - To provide beneficial guide for the application of the magnetic field in the bio treatment of the Cr(VI)-contained wastewater, sludge samples from the control bio system A (absent of magnetic field) and the contrast bio-system B (present of magnetic field) were used to adsorb the synthetic wastewater with 100 mg l(-1) Cr(VI). Influences of two adsorption modes, single adsorption and once continuous adsorption, on the Cr(VI) adsorption capacities of both sludge samples were compared. And the influence of regeneration on the Cr(VI) adsorption capacities were also studied. The results of adsorption experiments showed that the Cr(VI) adsorption capacities of the first single adsorption for sludge sample A and B were pretty nearly, which were 9.79 and 9.93 mg, respectively. And after 5 single adsorption periods, the total Cr(VI) adsorption capacity and efficiency of the sample B were 25.88 and 55.66 mg Cr(VI) g(-1)VSS, while those of the control were 14.95 and 33.98 mg Cr(VI) g(-1)VSS, respectively. For the sludge sample A and B after a single adsorption, both functions of regeneration were remarkable. But after 13 cycles of the single adsorption-regeneration, the Cr(VI) adsorption capacity and efficiency of the sample B were 110.15 and 189.91 mg Cr(VI) g( 1)VSS, while those of the control were 70.89 and 140.38 mg Cr(VI) g(-1)VSS, respectively. Though the Cr(VI) adsorption capacity of a once continuous adsorption period was more than that of a single adsorption period obviously, the Cr(VI) removal rates of the sludge sample A and B in the third period of once continuous adsorption-regeneration were only 8.12 and 33.51%, respectively. It was concluded that the weak magnetic field did improve the Cr(VI) bio-removal efficiency and the sludge stability, the batch treatment was an ideal operation mode for the bio-treatment of the Cr(VI)-contained wastewater, as compared with the continuous operation mode, but regeneration and enough sludge content were two necessary conditions to ensure the efficiency of batch treatment. PMID- 19554460 TI - Histopathology of Rhipicephalus sanguineus (Acari: Ixodidae) ticks fed on resistant hosts. AB - Histological features of Rhipicephalus sanguineus ticks fed on dog, a non resistant host, and on guinea pig, a resistant host, were compared. Unfed ticks and ticks from each host species were collected during first and third infestation and processed for histology. Many ticks from guinea pigs, especially during third infestation, were unattached, dehydrated and small. Only the midgut of ticks fed on guinea pigs had host leukocytes. Vacuolization of midgut cells was observed in all ticks, with exception of those fed on dogs for more than 96 h. Ticks of guinea pigs, particularly from third infestation, had vacuolated tracheae and swelling of malpighian tubules. Solely ticks from third infestation of guinea pigs displayed vacuolization of oocytes. Ticks fed on guinea pigs also had an increased number of guanine spherules. Observed alterations in ticks from guinea pigs are discussed. PMID- 19554461 TI - Early post-mortem changes and stages of decomposition in exposed cadavers. AB - Decomposition of an exposed cadaver is a continuous process, beginning at the moment of death and ending when the body is reduced to a dried skeleton. Traditional estimates of the period of time since death or post-mortem interval have been based on a series of grossly observable changes to the body, including livor mortis, algor mortis, rigor mortis and similar phenomena. These changes will be described briefly and their relative significance discussed. More recently, insects, mites and other arthropods have been increasingly used by law enforcement to provide an estimate of the post-mortem interval. Although the process of decomposition is continuous, it is useful to divide this into a series of five stages: Fresh, Bloated, Decay, Postdecay and Skeletal. Here these stages are characterized by physical parameters and related assemblages of arthropods, to provide a framework for consideration of the decomposition process and acarine relationships to the body. PMID- 19554462 TI - First detection of Rickettsia slovaca in Greece. AB - Seven hundred and three ticks collected in November and June of 2004-2006 from domestic animals in four localities of Halkidiki prefecture, Northern Greece, were tested for the presence of ricketsial DNA. Rickettsia slovaca was detected in one pool of Rhipicephalus bursa ticks, representing the first report of detection of R. slovaca in Greece. PMID- 19554463 TI - The life cycle of Amblyomma auricularium (Acari: Ixodidae) using rabbits (Oryctolagus cuniculus) as experimental host. AB - The life cycle of Amblyomma auricularium (Conil) is reported for the first time, using rabbits as experimental host. Developmental periods of free-living stages were observed in an incubator at 27 +/- 1 degrees C, 80 +/- 10% RH and 24 h darkness. The complete life cycle, including pre-feeding periods for each parasitic stage, ranged from 97 to 162 days. The overall sex ratio was 1.16:1 (M:F). Feeding and premolt periods, molting success, and engorgement weight of nymphs were statistically different between males and females (P < 0.01), but because their ranges overlapped, they cannot be used to predict the sex with accuracy. The potential role of rabbits as experimental hosts for rearing A. auricularium in the laboratory is discussed. PMID- 19554464 TI - Effect of high sulfate concentration on the corrosivity: a case study from groundwater in Harran Plain, Turkey. AB - Corrosion, which tends to increase the concentrations of certain metals in tap water, is one of the most important water quality problems as it can affect public health and public acceptance of water supply and the cost of providing safe water. In this context, this study aimed at investigating the scale formation tendency or corrosivity of groundwater in the semi-arid Harran Plain. The degree of scale formation tendency/corrosivity of water was determined considering pHs, Langelier Index, and Ryznar Index of groundwater samples. Except for well no.4, which is close to a local hot spring, all the wells had corrosive characteristics. The amount of CO(2) from the soil zone respiration and high sulfate concentration in the wells are important factors affecting corrosiveness. Results showed that precipitation, excessive irrigation, and change in groundwater level caused seasonal variation in corrosive characteristics. PMID- 19554466 TI - A serologic investigation for Peste des petits ruminants infection in sheep, cattle and camels (Camelus dromedarius) in Aydin province, West Anatolia. PMID- 19554465 TI - Synthesis, in vitro antitubercular activity and 3D-QSAR study of 1,4 dihydropyridines. AB - In continuation of our research program on new antitubercular agents, this article is a report of the synthesis of 97 various symmetrical, unsymmetrical, and N-substituted 1,4-dihydropyridines. The synthesized molecules were tested for their activity against M. tuberculosis H (37)Rv strain with rifampin as the standard drug. The percentage inhibition was found in the range 3-93%. In an effort to understand the relationship between structure and activity, 3D-QSAR studies were also carried out on a subset that is representative of the molecules synthesized. For the generation of the QSAR models, a training set of 35 diverse molecules representing the synthesized molecules was utilized. The molecules were aligned using the atom-fit technique. The CoMFA and CoMSIA models generated on the molecules aligned by the atom-fit method show a correlation coefficient (r (2)) of 0.98 and 0.95 with cross-validated r (2)(q (2)) of 0.56 and 0.62, respectively. The 3D-QSAR models were externally validated against a test set of 19 molecules (aligned previously with the training set) for which the predictive r(2)(r(r)(pred)) is recorded as 0.74 and 0.69 for the CoMFA and CoMSIA models, respectively. The models were checked for chance correlation through y scrambling. The QSAR models revealed the importance of the conformational flexibility of the substituents in antitubercular activity. PMID- 19554467 TI - Assessment of the nutritional status of indigenous scavenging chickens in Ada'a district, Ethiopia. AB - The nutritive values of scavenging feed resource bases and effects of season and chickens age on the latter were studied in smallholder farms in Aad'a, Ethiopia. The study included 210 households and 208 chickens. The mean weight of crop contents in all age groups ranged from 26.2 to 28.2 g, while it was 29.8 g and 24.7 g in the harvesting and non-harvesting seasons, respectively. Grains represented 48-49% of the mean weight of crop contents in all age groups and it was significantly higher (P < 0.05) during the harvesting season than the non harvesting period in both age groups. Kitchen wastes were next in abundance (26 29%) and were significantly more abundant in non-harvesting season in growers only and in hens than in growers (P < 0.001). The dry matter, crude protein, crude fiber, calcium, phosphorous and metabolizable energy levels of crop contents were 91.1-92.5%, 12.9-15.5%, 4.17-7.07%, 0.43-0.90%, 0.28-0.38% and 3404.3-3636.2 Kcal, respectively. Crude protein, calcium and phosphorus levels were below the requirements for egg production and growth. The scavenging feed resource base was critically deficient in these nutrients during the harvesting season. As these nutrients are vital for production, supplementary feeds rich in these nutrients would probably result in increased egg production and optimum growth. PMID- 19554468 TI - Repeated exposures to UVB induce differentiation rather than senescence of human keratinocytes lacking p16(INK-4A). AB - Skin cancers and extrinsic aging are delayed consequences of cumulative UV radiation insults. Exposure of human keratinocytes to UVB has been previously shown to trigger premature senescence. In order to explore the involvement of the cyclin-dependent kinase inhibitor p16(INK-4a) in UVB-induced premature senescence, we developed an original model of repeated sublethal exposures of human keratinocytes deficient in p16(INK-4a). We did not observe any significant increase of senescence-associated beta-galactosidase activity positive cells following UVB exposure in this cell line in contrast to primary keratinocytes, suggesting a role for p16(INK-4a) in UVB-induced senescence. However, we detected sustained DNA damage, prolonged cell cycle arrest, and induction of markers of epidermal differentiation like involucrin and filaggrin as consequences of the repeated exposures. Keratinocytes exposed to the same dose of UVB in a single exposure died. Furthermore, the abundance of the keratins 6, 16 and 17 was increased in keratinocytes exposed repeatedly to UVB suggesting an alternative differentiation. This model allows the induction of a state of differentiation observed in vivo with differentiation uncoupled from premature senescence. PMID- 19554469 TI - Acetylcholine and muscarinic receptor function in cerebral cortex of diabetic young and old male Wistar rats and the role of muscarinic receptors in calcium release from pancreatic islets. AB - We investigated acetylcholine esterase (AChE) activity, acetylcholine and muscarinic M1, M3 receptors kinetics in the cerebral cortex of young and old streptozotocin induced and insulin treated diabetic rats. The role of muscarinic receptors in intracellular calcium release from pancreatic islets was studied in vitro. Wistar rats of 7 and 90-weeks old were used. All studies were done in cerebral cortex. AChE assay was done by spectrophotometric method. Radioreceptor binding assays were done for Acetylcholine, Muscarinic M1 and M3 receptors using specific ligands. Calcium imaging was done using fluo4-AM in pancreatic cells. Ninety-weeks old control rats showed significantly decreased Vmax and increased Km for AChE compared to 7-weeks old control rats. An increased Vmax observed in both 7 and 90-weeks old diabetic groups with significant decrease in Km. Scatchard analysis using specific agonists showed significant decrease in the B (max) and K (d) of acetylcholine and muscarinic M1 receptors in 90-weeks old control rats compared to 7-weeks old control. Binding studies for M3 receptors showed no significant change compared to 7-weeks old control. Acetylcholine, muscarinic M1 and M3 receptor number significantly increased in 90-weeks old diabetic rat groups compared to their respective controls. Insulin treatment significantly reversed the binding parameters to near control compared to diabetic group. In vitro studies showed that acetylcholine through muscarinic M1 and M3 receptors' stimulated calcium release from the pancreatic islets. Thus our studies suggest that Insulin signaling play an important part in differentially regulating pancreatic cholinergic activity, and the diabetes mediated cortical dysfunctions with age. PMID- 19554470 TI - The changing roles of pharmacists in community pharmacies: perception of reality in India. AB - AIM OF THE REVIEW: To summarise the state of community pharmacy in India including: the provision of patient care, pharmacy education, the pharmaceutical market, healthcare delivery, community pharmacy services, the professional role of community pharmacists, and future developments. METHOD: Medline, Scirus, and Google Scholar databases and the journals "Indian Journal of Pharmaceutical Sciences" and "Indian Journal of Hospital Pharmacy" were searched up to the end of December 2008. In addition to these, other available sources were used to identify relevant articles. RESULTS: India has a fast growing pharmaceutical industry sector and a need for well educated pharmacists. Domestic sale of medicines is estimated to be $5 billion during 2006 and increasing. The supply of medicines to the population is undertaken by privately owned community pharmacies and sometimes also by hospital pharmacies. Community pharmacists are involved only in dispensing medicines. Community pharmacists have an opportunity to improve healthcare of the population, particularly of the disadvantaged section of the society that does not have the resources to visit clinics (both the poor and rural population). However, important barriers to the provision of pharmaceutical care exists, including lack of proper education and training of pharmacists, weak implementation of existing laws, and lack of recognition of the pharmacy as a profession by the other healthcare professionals. CONCLUSION: The healthcare services in community pharmacies, currently insignificant, must undergo reforms to meet the changing needs of modern medicines users. The pharmacist's role in patient care is expected to grow as professional and educational standards develop. Although pharmacists' contributions to health care are not yet recognized, there is every reason to be optimistic toward making patient care in community pharmacy setting a success. For this, the educational system for pharmacists has to be adapted. PMID- 19554471 TI - Impact of a computerized physician order entry system on compliance with prescription accuracy requirements. AB - OBJECTIVE: To assess the change in non-compliant items in prescription orders following the implementation of a computerized physician order entry (CPOE) system named PreDiMed. SETTING: The department of internal medicine (39 and 38 beds) in two regional hospitals in Canton Vaud, Switzerland. METHOD: The prescription lines in 100 pre- and 100 post-implementation patients' files were classified according to three modes of administration (medicines for oral or other non-parenteral uses; medicines administered parenterally or via nasogastric tube; pro re nata (PRN), as needed) and analyzed for a number of relevant variables constitutive of medical prescriptions. MAIN OUTCOME MEASURE: The monitored variables depended on the pharmaceutical category and included mainly name of medicine, pharmaceutical form, posology and route of administration, diluting solution, flow rate and identification of prescriber. RESULTS: In 2,099 prescription lines, the total number of non-compliant items was 2,265 before CPOE implementation, or 1.079 non-compliant items per line. Two-thirds of these were due to missing information, and the remaining third to incomplete information. In 2,074 prescription lines post-CPOE implementation, the number of non-compliant items had decreased to 221, or 0.107 non-compliant item per line, a dramatic 10 fold decrease (chi(2) = 4615; P < 10(-6)). Limitations of the computerized system were the risk for erroneous items in some non-prefilled fields and ambiguity due to a field with doses shown on commercial products. CONCLUSION: The deployment of PreDiMed in two departments of internal medicine has led to a major improvement in formal aspects of physicians' prescriptions. Some limitations of the first version of PreDiMed were unveiled and are being corrected. PMID- 19554472 TI - Some possible dynamical constraints for life's origin. AB - Oscillating biochemical reactions are common in cell dynamics and could be closely related to the emergence of the life phenomenon itself. In this work, we study the dynamical features of some classical chemical or biochemical oscillators where the effect of cell volume changes is explicitly considered. Such analysis enables us to find some general conditions about the cell membrane to preserve such oscillatory patterns, of possible relevance to hypothetical primitive cells in which these structures first appeared. PMID- 19554473 TI - Efficient amplification of light and heavy chain variable regions and construction of a non-immune phage scFv library. AB - Non-immune phage scFv library is one of the most attractive resources for therapeutics, diagnostics and basic research. As a matter of fact, quality of the library is limited by inefficient PCR cloning of antibody genes using degenerated primers. PCR using this type of primers is difficult to optimize conditions for efficient amplification, and therefore causes loss of antibody diversities. To overcome this problem, we described a novel two-step amplification of V(kappa) and V(H) genes with newly designed primer sets. Initially, we amplified V(kappa) and V(H) genes from their signal sequences to the joining region to keep antibody diversity as large as possible. Thereafter, highly degenerated primers were used to amplify the V(kappa) and V(H) genes from the framework region 1 to the joining region. The V(kappa) and V(H) genes from the second PCR then were linked by PCR overlapping extension to generate the scFv library. Fifteen clones from the library were randomly picked and sequenced, and the diversity of full-length scFvs was confirmed. Expression capability of clones in the library was 80% after confirmation using colony hybridization. The results demonstrated the efficiency of this strategy and the primer sets for construction of the scFv library. PMID- 19554474 TI - Confirmatory and exploratory factor analysis of the caregiver quality of life index-cancer with Turkish samples. AB - PURPOSE: The purpose of this study was to test the reliability and validity of the Turkish version of the CQOLC in Turkey. METHODS: The 35-item English version of the CQOLC was translated into Turkish following the standard translation methodology. The questionnaire was administered to 237 caregivers of patients with cancer. Confirmatory and exploratory factor analyses (CFA and EFA) were carried out using principal component analysis with varimax rotation and Kaiser Normalization to test its construct validity. We used Cronbach's alpha to examine the CQOLC's reliability (internal consistency). RESULTS: The CFA did not confirm the original factor model. The EFA yielded a 25-item measure with a four-factor solution with different labels for three of the four original scales (shown in parentheses): Psychological Distress (Burden), Disruption in Daily Life (Disruptiveness), Caregiving Responsibility (Positive Adaptation), and Financial Concerns (Financial Concerns). Cronbach's alpha for the total scale was 0.88 and subscale alpha coefficients ranged from 0.73 to 0.83. CONCLUSIONS: The results indicate some differences in the factor structures of the CQOLC scale between Turkish and American samples but provided preliminary support for the Turkish version of the CQOLC as a reliable and valid measure of the quality of life of Turkish cancer caregivers. PMID- 19554476 TI - Childhood genetic testing for familial cancer: should adoption make a difference? AB - Professional guidelines and practice in clinical genetics generally counsel against predictive genetic testing in childhood. A genetic test should not be performed in a child who is too young to choose it for himself unless that test is diagnostic, will lead to an intervention to prevent illness, or enable screening. It is therefore generally considered unacceptable to test young children for adult-onset cancer syndromes. However, these guidelines are challenged when clinical genetics services receive requests from adoption agencies or pre-adoptive parents for predictive genetic tests in children being placed for adoption. Testing will foreclose a pre-adoptive child's future autonomous right to choose, yet those commissioning these tests argue that adoption should form a special case. In this paper, we argue that predictive genetic testing as part of a pre-adoptive 'work-up' should be discouraged when the same test would not generally be carried out in a child who is not being adopted. We present an argument based on a principle of consistency and question those claims that privilege the adoptive process, whilst acknowledging the array of uncertainties faced by pre-adoptive parents. We suggest that if pre-adoptive testing is considered, this should only take place after prospective adoptive parents have had the opportunity to meet the clinical genetics team and fully understand the implications of the testing process. PMID- 19554475 TI - Conflict between values and technology: perceptions of preimplantation genetic diagnosis among women at increased risk for hereditary breast and ovarian cancer. AB - Members of families affected by hereditary cancer are often concerned about passing on risk to offspring. Preimplantation genetic diagnosis is a procedure performed to identify embryos that inherit mutations placing them at risk for hereditary conditions. Little is known about attitudes toward the use of this technology among individuals at risk for hereditary breast and ovarian cancer. We sought to determine high risk women's attitudes. This study is a qualitative examination of comments from women who participated in an online survey regarding knowledge and attitudes of preimplantation genetic diagnosis among individuals affected by hereditary breast and ovarian cancer. More than half the respondents held less favorable attitudes about the use of preimplantation genetic diagnosis for hereditary breast and ovarian cancer for both themselves and others. However, among the women who felt favorable about its usage, the majority said it became a new option for them to pursue parenthood whereas previously they had opted to not have a biological child. The high percentage of respondents who have never heard of preimplantation genetic diagnosis and who were in favor of this technology for hereditary breast and ovarian cancer indicates the need for educational campaigns to increase awareness and provide information about the procedure, access and affordability. Further research is needed to determine how this population would like this information presented to them and how best to instruct health care professionals to present this topic to women who do not know to ask about it. PMID- 19554477 TI - Iterated function systems in the hippocampal CA1. AB - How does the information of spatiotemporal sequence stemming from the hippocampal CA3 area affect the postsynaptic membrane potentials of the hippocampal CA1 neurons? In a recent study, we observed hierarchical clusters of the distribution of membrane potentials of CA1 neurons, arranged according to the history of input sequences (Fukushima et al Cogn Neurodyn 1(4):305-316, 2007). In the present paper, we deal with the dynamical mechanism generating such a hierarchical distribution. The recording data were investigated using return map analysis. We also deal with a collective behavior at population level, using a reconstructed multi-cell recording data set. At both individual cell and population levels, a return map of the response sequence of CA1 pyramidal cells was well approximated by a set of contractive affine transformations, where the transformations represent self-organized rules by which the input pattern sequences are encoded. These findings provide direct evidence that the information of temporal sequences generated in CA3 can be self-similarly represented in the membrane potentials of CA1 pyramidal cells. PMID- 19554478 TI - Comparison of the cardiac electrophysiology and general toxicology of two formulations of intravenous amiodarone in dogs. AB - Intravenous amiodarone (AIV) must be administered slowly after dilution to avoid hypotension, which is due to the cosolvents polysorbate 80 and benzyl alcohol used in its formulation. PM101 is a formulation of amiodarone devoid of these cosolvents, which enables bolus administration. We evaluated any potential toxicity or exaggerated adverse cardiac electrophysiologic effects of PM101 compared with AIV and control. Beagle dogs were treated with the human-equivalent amiodarone loading dose (2.14 mg/kg) with PM101 (bolus push) or AIV (10 min infusion in the toxicology study and bolus push in the electrophysiology study) followed by maintenance infusion (0.014 mg kg(-1) min(-1) through 6 h followed by 0.007 mg kg(-1) min(-1) through 14 days) or a control. General toxicology was assessed in conscious dogs over 14 days. Cardiac electrophysiology was assessed in a separate cohort of anesthetized dogs during the first 20 min of dosing. In the toxicology study, dosing in all animals in the AIV group was terminated within 17 min of initiation due to a severe hypersensitivity reaction. There were no acute adverse clinical signs in the PM101 or control groups. There were no significant effects on body weight or ECG parameters, and no adverse histomorphologic changes were seen in dogs that received PM101 or AIV. No significant exaggerated cardiac electrophysiologic effects of the approved doses PM101 or AIV were observed. PM101 may represent a formulation of intravenous amiodarone that could be administered rapidly without dilution in the setting of life-threatening cardiac arrhythmias. PMID- 19554479 TI - Biliary-bronchial fistula following radiation therapy for gall bladder cancer successfully treated by endoscopic therapy. PMID- 19554480 TI - Vaccination for atherosclerosis. AB - Atherosclerosis is an inflammatory disease, and several antigens have been shown to activate the immune response and affect the development of atherogenesis. This suggests that modulation of the immune system could represent a useful approach to prevent and/or treat this disorder. A vaccination approach might be a useful, effective tool in the modern arsenal of cardiovascular therapy and could possibly be used on a large scale at a low cost. Several modalities of vaccines have been tested against lipoproteins, cholesterol, molecules involved in cholesterol metabolism, atherosclerosis-associated microorganisms, and other molecules (heat shock protein, CD99, vascular endothelial growth factor-receptor, interleukin-2), with promising results. Nevertheless, a deeper understanding of the role of immunization in atherosclerosis will be essential to the use of vaccines in clinical medicine. PMID- 19554481 TI - The premenstrual syndrome and fibromyalgia--similarities and common features. AB - The aim of the study was to assess the clinical similarities and common features of fibromyalgia syndrome (FM) and premenstrual dysphoric syndrome (PMDD). Thirty young patients who met the diagnostic criteria for PMDD were included in the study and compared to 26 women belonging to the medical staff of a general psychiatry department. All enrollees were interviewed and examined by a skilled physician. They completed the following nine survey items: demographic information, clinical health assessment questionnaire, fibromyalgia impact questionnaire, sleep and fatigue questionnaires, Sheehan disability scales, SF-36 assessment for quality of life, visual analog scale for pain, Mini International Neuropsychiatric Interview (MINI) questionnaire (assessment of coexistent psychiatric conditions), and the premenstrual severity scale. Additionally, each individual underwent a physical examination measuring the classical tender points and was asked to describe the distribution and continuum of her pain or tenderness. The PMDD group scored significantly higher in the measures pain and tenderness as well as in severity of premenstrual symptoms compared to the control group. Five patients in the PMDD group and none in the control group had FM. Quality of life measured by the SF-36 was higher in the control group than in the PMDD group and correlated with the degree of tenderness reported. Psychiatric comorbidity was significantly more common in the PMDD group, affecting 16 of the 30 PMDD patients compared to only three of the 26 control patients. In this study, patients with PMDD were found to have higher levels of tenderness, higher psychiatric comorbidity, greater level of physical disabilities, and a lower quality of life. These parameters were highly correlated with a lower pain threshold. PMID- 19554482 TI - NAD+ availability and proteotoxicity. AB - It has been shown that NAD(+) availability is important for neuronal survival following ischemia (Liu et al., Neuromolecular Med 11:28-42, 2009). It is proposed here that NAD(+) may also control proteotoxicity by influencing both formation and catabolism of altered proteins. It is suggested that low NAD(+) availability promotes synthesis of methylglyoxal (MG) which can induce formation of glycated proteins, ROS, and dysfunctional mitochondria. That glyoxalase overexpression and carnosine are both protective against MG and ischemic injury support this proposal. Recognition and elimination of altered proteins is enhanced by NAD(+) through effects on stress protein expression and autophagy. PMID- 19554484 TI - Downregulation of ABI1 expression affects the progression and prognosis of human gastric carcinoma. AB - Abelson interactor protein-1 (ABI1) is a promising candidate tumor suppressor, and plays critical roles both in the pathogenesis of BCR-Abl-induced leukemia and in the spread of several solid tumors. The expression of ABI1 and its role in cancer progression and prognosis are largely unknown in the majority of solid tumors, including gastric cancer. In this study, we analyzed the correlation between ABI1 expression and the clinicopathological characteristics, tumor progression, and prognosis of patients with gastric carcinoma. Tissue specimens were from 103 gastric cancer patients who underwent gastrectomy in our hospital between January 2000 and December 2007. Among them 59 tumor tissue samples were matched with normal tissue samples. The expression of ABI1 protein was measured using immunohistochemical staining of paraffin-embedded tissue specimens. Meanwhile, quantitative real-time RT-PCR and Western blotting were used to identify the expression of ABI1 in human gastric normal mucosal cell line (GES-1) and gastric cancer cell lines (N87, AGS). We performed a statistical analysis of the potential correlation between ABI1 expression and the patients' clinicopathological characteristics, 5-year survival, and median survival time. The immunohistochemical staining results of 59 patients showed that ABI1 was expressed in 28.8% (17/59) of gastric cancer tissues, compared to 91.5% (54/59) of normal samples. ABI1 expression in 103 patients was strongly correlated with tumor differentiation, clinical stage, and lymph node status (P < 0.01). The 5 year survival rate was 15.3% in the ABI1-negative group and 63.7% in the ABI1 positive group. Median survival time in the ABI1-negative and ABI1-positive groups was 25.0 months (95% CI: 19.7-30.3) and 74.0 months (95% CI: 54.6-93.3), respectively. There was a significant difference between the two groups (chi(2) = 10.888, P = 0.001). Furthermore, we found that ABI1 expressed lowly in poor differentiated AGS, whereas highly in GES-1 and well-differentiated N87. Downregulation of ABI1 expression in human gastric carcinoma may play a critical role in tumor progression and in determining patient prognosis. ABI1 may be a useful diagnostic or prognostic molecular biomarker, and might be a potential target for therapeutic intervention. PMID- 19554483 TI - Olfactomedin domain-containing proteins: possible mechanisms of action and functions in normal development and pathology. AB - A family of olfactomedin domain-containing proteins consists of at least 13 members in mammals. Although the first protein belonging to this family, olfactomedin, was isolated and partially characterized from frog olfactory neuroepithelim almost 20 years ago, the functions of many family members remain elusive. Most of the olfactomedin domain-containing proteins, similar to frog olfactomedin, are secreted glycoproteins that demonstrate specific expression patterns. Other family members are membrane-bound proteins that may serve as receptors. More than half of the olfactomedin domain-containing genes are expressed in neural tissues. Data obtained over the last several years demonstrate that olfactomedin domain-containing proteins play important roles in neurogenesis, neural crest formation, dorsal ventral patterning, cell-cell adhesion, cell cycle regulation, and tumorigenesis and may serve as modulators of critical signaling pathways (Wnt, bone morphogenic protein). Mutations in two genes encoding myocilin and olfactomedin 2 were implicated in glaucoma, and a growing number of evidence indicate that other genes belonging to the family of olfactomedin domain-containing proteins may contribute to different human disorders including psychiatric disorders. In this review, we summarize recent advances in understanding the possible roles of these proteins with special emphasis on the proteins that are preferentially expressed and function in neural tissues. PMID- 19554485 TI - Use of linezolid in critically ill patients admitted to intensive care units. AB - All indications of linezolid (LZD) in Intensive Care Units (ICU) were included as cases in an observational, prospective and multicentre study. One hundred thirty nine indications were analyzed. In most cases (92.7%), treatment for nosocomial infections was indicated. The most frequent infection was pneumonia (42.7%), followed by catheter-related bacteraemias (CRB). A total of 58.7% of the indications were empirical and in 45.7% of the cases the cultures confirmed infection by gram-positive cocci (GPC). In 43 cases (31.2%), the indication was made as a rescue measure (mainly due to clinical failure) in patients previously treated with glycopeptides. Of isolated GPC, 70.2% were methicillin-resistant. The cure rate of the population per intent-to-treat was 73.2%. Only one case of thrombocytopenia was recorded. Conclusions. LZD is used with a high degree of diagnostic safety. In the ICU, it is primarily indicated to treat pneumonias and CRB with good clinical and microbiological response. This antibiotic has acted as a good therapeutic resource against clinical failure in infections treated with glycopeptides. PMID- 19554486 TI - [Multicenter study of in vitro activity of tigecycline in clinical isolates from 30 centers in Spain]. AB - INTRODUCTION: In the last years, the increase of antibiotic resistances of gram positive and gram-negative bacteria is an important therapeutic problem. The antimicrobial activity of tigecycline, a novel glycylcycline, was evaluated against 750 bacterial isolates from 30 centers in Spain. METHODS: Multicenter and retrospective study. In 2005, thirty laboratories participated in this study. Data collected in this study included antimicrobial susceptibilities for S.aureus resistant to methicillin (MRSA), ESBL- E. coli or ESBL- K. pneumoniae, E. coli resistant to quinolons (E.coli- QR), Klebsiella spp and E. faecalis. Trains were obtained of the each Hospital s collection (5 strains of each microorganisms). The susceptibility determinations were performed locally by each laboratory following the standard method usually performed. The tigecycline susceptibility determinations were performed with E/test. RESULTS: Tigecycline was the most potent agent against MRSA, E. faecalis, E.coli-QR and ESBLs enterobacteriaceae; with MIC50-MIC90 values of: 0.125-0.25 g/ml; 0.125-0.5 g/ml; 0.25-0.75 g/ml and 0.38-1.5 g/ml; respectively. CONCLUSIONS: The results of this study confirm the excellent in vitro activity of tigecycline against gram-positive and gram negative pathogens, including multirresistant microorganisms. PMID- 19554492 TI - [Criteria of sleep quality]. PMID- 19554489 TI - [Recommendations of the Heart Diagnosis Working Group of the German Roentgen Society for use of computerized tomography and magnetic resonance tomography in heart diagnosis. 1-- Computerized tomography]. AB - MRI and CT have become important tools in the diagnosis of cardiac diseases. The permanent technical development and the continuously growing clinical and scientific experience required an update of the previous recommendations regarding the spectrum of clinical applications and the technical aspects of MRI and CT of the heart. The Working group Herzdiagnostik of the German Roentgen Society has revised its recommendations based on the current literature and has updated technical requirements and clinical indications. Part I of the recommendations deals with cardiac computed tomography while part II focuses on MRI in cardiac applications. These recommendations shall serve as a guideline when deciding on appropriate technique for non-invasive cardiac imaging and shall help to perform high quality imaging based on a standardised approach. PMID- 19554493 TI - [Pain processing, pain memory and quality of sleep]. PMID- 19554494 TI - [Significance of sleep quality for the patient with chronic pain]. PMID- 19554495 TI - [Optimizing the galenical formulation of analgetics for the treatment of chronic pain conditions: possible effects on sleep disorders]. PMID- 19554496 TI - [Practice-oriented treatment of pain and sleep deficit]. PMID- 19554497 TI - [Aspects of health economics in the treatment of pain and sleep deficit]. PMID- 19554498 TI - [Sleep quality, pain therapy. Summing up for the user]. PMID- 19554499 TI - [Goal analysis and goal operationalisation: a group intervention for the enhancement of work motivation]. AB - BACKGROUND: Work motivation, mental well-being and competencies of self regulation are linked to successful job-related reintegration after rehabilitation. Based on the Diagnostical Instrument to assess Work motivation (Diagnostikinstrument fur Arbeitsmotivation DIAMO) and existing training programs, a new group intervention, the goal analysis and goal operationalization, was developed and evaluated. The objective of this intervention, designed for participants of a rehabilitation program was to enhance work motivation and volitional control processes (self-regulation and self-control), to encourage job-related goal orientation and to thereby increase the probability of goal achievement. PATIENTS AND METHODS: In a quasi experimental longitudinal design 207 patients (111 experimental group/96 control group) were tested. The experimental group took part in the job-related training (ZAZO) in addition to the usual rehabilitation. The evaluation was conducted through various scales at t0 (beginning) and t1 (end of the training). Scales for the measurement of work motivation, mental well-being, status of rehabilitation, competencies of self-regulation and the subjective prognosis of the ability to work were used. As direct effects of the training an enhancement of work motivation and of an improved subjective prognosis of the ability to work were expected. Accordingly, a positive influence on the subjective well-being as indirect effects, were anticipated in the long run, the experimental group should also show an enhanced job-related reintegration. RESULTS: Participants of the experimental group showed significantly higher values on particular scales of the Diagnostical Instrument of Work motivation as opposed to the control group (curiosity motive, attitudes to work and contact motive). Most notably, significant interactional effects could be found on the scale for the subjective prognosis of the ability to work, which is a highly reliable instrument and important predictor for prospective job-related reintegration. The training ZAZO led to an enhancement of work motivation and subjective prognosis of the ability to work, both having positive effects on job-related orientation. CONCLUSIONS: First evaluation results of the training ZAZO document the positive effect on both work motivation and subjective prognosis of the ability to work. The results of the evaluation present the good acceptance of the participants (93.3% = rather content to very content). The implementation of the group intervention in existing programs of rehabilitation can benefit job-related reintegration and will thereby contribute to lower costs for pension regulatory authorities in the long run. PMID- 19554500 TI - [Women with a history of pregnancy loss or abortion in a behavioural medicine hospital--an exploratory field study]. AB - OBJECTIVE: Gaining insights into the frequencies of negative obstetric histories in inpatients of a behavioural medicine hospital and the extent of their current psychological burdens. METHOD: We acquired sociodemographic, obstetric and psychological (psychopathology, coping patterns, and attribution styles) data of 117 inpatients by means of standardized questionnaires and interviews. RESULTS: Eighteen percent of the women exhibited a negative obstetric history (spontaneous, medically indicated or voluntary abortion) that dated back an average of 16 years. These women suffered more often from somatoform disorders and showed higher depression scores as well as other distinctive problems regarding their psychological wellbeing and their attributional and coping styles. Women who had undergone voluntary abortions showed a particular pattern of psychological distress. Those women who still felt psychological burden of pregnancy loss or abortion were younger and had fewer children. CONCLUSIONS: Almost one fifth of this inpatient sample in a behavioural medicine hospital exhibited a negative obstetric history and one third of these women still were affected psychologically. Clinicians therefore need to include obstetric data when taking a woman's psychiatric history, in particular with regard to depression, in order to be able to include this issue in their treatment plan, even if the pregnancy loss or abortion dates back many years. PMID- 19554501 TI - [Long-term school-development of children with impaired language skills: 15-years follow up after dismissal from child care centres with speech therapy]. AB - BACKGROUND: Severe language impairment (SLI) in children causes multiple developmental disturbances. Less is known concerning the long-term efficacy of Child care centres with speech therapy (CCC) and the validity of of IQ-testing. AIM OF THE STUDY: Follow-up of school-development of SLI-children from CCC and analysis of the the prognostic relevance of IQ-testing as monitored by the surrogate-parameter "school-development". METHODS: 1. Retrospective study with questionnaires; 2. Analysis of documents from CCC. 88 children (60 males, 28 females) were included. RESULTS: Median length of stay in CCC was 18 months (range: 12-36 months); present study was performed 0,5 to 15 years after dismissal from CCC. Due to parents;remembrance more than 90% of the children suffered on symptoms of SLI within the 2 (nd) year of life; about 30% of the children received grommets; IQ was determined by SON-R 2,5-7 in 68 children and amounted 99 (58-131). In 32 children (>10 years) school-careers were followed after termination of primary schools: 5 children visited "schools for handicapped children", 26 children attended mainstream-schools, 19 of them in schools with higher academic levels ("advanced-" and "integrated high schools", "grammar schools"). In 16 of 68 children the IQ amounted between 70 to 89, 11 of these children visited a mainstream school. In 45 of these children IQ was >90, 6 of these children had to attend a "school for handicapped children". CONCLUSIONS: IQ testing is thought to be a limited prognostic parameter for long-term school development of SLI-children. Findings indicate, that about 80% of the SLI children benefitted by the therapy in specialized CCC and visited a mainstream school. PMID- 19554502 TI - [Modification of the mineralizing capacity of a saliva substitute (saliva natura) on enamel in vitro]. AB - PURPOSE: This in vitro study investigated the effects of exposure to modified (with respect to calcium phosphate saturation) solutions of a commercial available saliva substitute (Saliva natura) on mineralization of enamel in vitro. METHODS: Bovine enamel specimens were prepared. Before and after demineralization (pH 4.95, 14 d, 37 degrees C), one-quarter of each specimen's surface was covered with nail varnish (control of sound/demineralized tissue). Specimens were exposed either to original Saliva natura [saturation with respect to octacalciumphosphate (S (OCP)): 0.03, pH 5.8] or to three lab-produced Saliva natura modifications (S (OCP): 1, 2, 3, pH 6.0) for two and five weeks (37 degrees C). An aqueous solution (S (OCP): 2.7, pH 7.0; Buskes remineralizing solution) served as positive control. Differences in mineral losses (DeltaDeltaZ) and lesion depths (DeltaLD) before and after storage were evaluated from microradiographs. RESULTS: After two weeks storage no differences among the solutions with regard to DeltaDeltaZ and DeltaDeltaLD could be observed (p>0.05; ANOVA). Five weeks storage in original Saliva natura resulted in significantly lower DeltaDeltaZ values compared to all other solutions (p<0.05). No differences with respect to DeltaDeltaZ among the modified solutions (S (OCP) 1, 2, 3; p>0.05) could be observed, whereas storage in the remineralizing solution resulted in higher DeltaDeltaZ values compared to all other solutions (p<0.05). For DeltaLD similar results could be revealed. However, no differences between the remineralizing solution and Saliva natura S (OCP) 2 could be shown (p>0.05). CONCLUSION: Saliva natura with an S (OCP) of 2 showed the highest remineralizing capacities. More pronounced remineralization could not be achieved with a higher S (OCP) of 3 under the conditions chosen. PMID- 19554503 TI - [The topical application of mitomycin C in the treatment of scar formation and stenosis in hollow organs of the head and neck: a field report]. AB - INTRODUCTION: The antibiotic und antitumoral effect of Mitomycin C (MMC), a derivative of Streptomyces caespitosus, has been known since 1956. Besides its use as an adjunction in the treatment of breast, lung and prostate cancer, or as a second-line cytostatic drug for head and neck squamous cell carcinoma (HNSCC), since 1963, MMC has also been successfully used in the suppression of post operative scar formation, particularly in the field of ophthalmology. This is due to its modulation of fibroblast activity, which enables decreased scarring and fibrosis. In this resume, we wish to recapitulate our long years of experience in the topical application of Mitomycin C in the treatment of scar formation and stenosis in head and neck organs. PATIENTS AND METHODS: A retrospective analysis on the basis of clinical disease courses and findings (image documentation, questionnaires, pulmonary function tests) covering an examination period of 10 years, was performed on 40 patients with stenosising lesions and a mean age of 54 years. The fields of application included laryngeal, tracheal, oesophageal stenosis and stenosis of the external ear canal and the choane. RESULTS: After combined application of MMC and surgical intervention in cases of recurrent stenosising processes in head and neck organs, especially the larynx and the trachea, a sustained improvement was achieved in the pre-operative stenosis level as well as in the pre-operative, severely limited, forced inspiratory volume in 1 second (F1V1). CONCLUSION: The topical application of MMC appears to be an effective adjunction as a concept of treatment for stenosising, scar-forming lesions. This topical application, however, is not a substitute for correct diagnosis and appropriate surgical treatment. It must be regarded as a purely adjunctive manoeuvre. During the 10 years in which it was used, the clinical findings confirmed an enhancement in the containment of complex cases without the occurrence of any complications. PMID- 19554504 TI - Expression of hexokinases and glucose transporters in treated and untreated oesophageal adenocarcinoma. AB - The aim of this study was to assess the expression pattern of the high glucose affinity glucose transporters GLUT 1, 2, 3, 4, 8 and 9 and of hexokinases I, II and III in newly diagnosed oesophageal adenocarcinoma by means of immunohistochemistry. Twenty patients eligible to undergo primary surgery and 18 patients with incomplete pathological response following induction radio chemotherapy, all suffering from oesophageal adenocarcinoma, were included in the study. The intensity and amount of positive tumour cells in the immuno-reaction (histology score (Hscore)) for GLUT 1, 3, 4, 8 and 9 as well as for hexokinase I, II and III were assessed independently by two experienced observers, blinded to the clinical results. In patients that underwent primary surgery, Hscores of GLUT8 (micro 6.7; sd3.3) and GLUT1 (micro 5.5; sd: 5.3) were significantly higher than Hscores of GLUT9 (micro 2.2; sd 1.5) and GLUT3 (micro 3.2, sd: 2.5). Hscores of hexokinase I (micro : 8.3; sd: 4.3), II (micro 5.5, sd: 4.0) and III (I 1.5, sd: 0.7) were all significantly different from each other (p<0.04). In patients that underwent radio-chemotherapy prior to surgical tumour resection, micro Hscores were 6.9 (sd: 4.4) for GLUT1, 6.8 (sd: 5.3) for GLUT3, 5.9 (sd: 4.2) for GLUT8, 3.4 for GLUT9 (sd: 2.7) and 2.3 (sd: 3.6) for GLUT 4. Hscores of GLUT1 and GLUT3 were significantly higher than Hscores of GLUT4. Finally, Hscores of patients with radio-chemotherapy for GLUT3, hexokinase II and III were significantly higher when compared to patients that underwent primary surgery. PMID- 19554505 TI - Vaspin and amylin are expressed in human and rat placenta and regulated by nutritional status. AB - Amylin (islet amyloid polypeptide) and vaspin (visceral adipose tissue specific serpin) are gut and adipocyte hormones involved in the regulation of body weight homeostasis. The aim of this study was to examine whether amylin and vaspin are expressed in human and rat placenta, as well as their regulation by nutritional status. Our results demonstrate that amylin and vaspin are localized in both human and rat placenta. In the rat term placenta vaspin was demonstrated in the trophoblast of the fetal villi, the labyrinth. Vaspin immunostaining in human placenta was localized in cytotrophoblast and syncytiotrophoblast in the first trimester placentas while in the third trimester vaspin was localized in the syncytiotrophoblast. Regarding amylin, rat placenta of 16 days of gestational age showed an intense immunostaining, mainly localized in the labyrinth. On the other hand, in the human third trimester placenta amylin immunoreactivity was intense in the syncytiotrophoblast of the chorionic villi and in decidual cells. Furthermore, placental amylin and vaspin showed an opposite pattern of expression during pregnancy, with vaspin showing the highest expression level at the end and amylin at the beginning of pregnancy. Finally, food restriction also has contrary effects on their expression, increasing vaspin but decreasing amylin placental mRNA and protein levels. Taken together, our results demonstrate that vaspin and amylin are modulated by energy status in the placenta, which suggests that these proteins may be involved in the regulation of placental metabolic functions. PMID- 19554506 TI - Expression of eight genes of nuclear factor-kappa B pathway in multiple myeloma using bone marrow aspirates obtained at diagnosis. AB - PURPOSE: To evaluate the expression of NF-kappaB pathway genes in total bone marrow samples obtained from MM at diagnosis using real-time quantitative PCR and to evaluate its possible correlation with disease clinical features and survival. MATERIAL AND METHODS: Expression of eight genes related to NF-kappaB pathway (NFKB1, IKB, RANK, RANKL, OPG, IL6, VCAM1 and ICAM1) were studied in 53 bone marrow samples from newly diagnosed MM patients and in seven normal controls, using the Taqman system. Genes were considered overexpressed when tumor expression level was at least four times higher than that observed in normal samples. RESULTS: The percentages of overexpression of the eight genes were: NFKB1 0%, IKB 22.6%, RANK 15.1%, RANKL 31.3%, OPG 7.5%, IL6 39.6%, VCAM1 10% and ICAM1 26%. We found association between IL6 expression level and International Staging System (ISS) (p=0.01), meaning that MM patients with high ISS scores have more chance of overexpression of IL6. The mean value of ICAM1 relative expression was also associated with the ISS score (p=0.02). Regarding OS, cases with IL6 overexpression present worse evolution than cases with IL6 normal expression (p=0.04). CONCLUSION: We demonstrated that total bone marrow aspirates can be used as a source of material for gene expression studies in MM. In this context, we confirmed that IL6 overexpression was significantly associated with worse survival and we described that it is associated with high ISS scores. Also, ICAM1 was overexpressed in 26% of cases and its level was associated with ISS scores. PMID- 19554507 TI - Improved methodology for the detection and quantification of the acrosome reaction in mouse spermatozoa. AB - This study evaluates the use of two fluorescein-labelled (FITC) plant lectins, Pisum sativum (edible pea) agglutinin (PSA) and Arachis hypogaea (peanut) agglutinin (PNA), in order to determine the most accurate and reliable method to experimentally detect and assess the acrosome reaction in mouse spermatozoa. PNA FITC labelling was restricted to the acrosome and was not influenced by the fixation procedure; either absolute methanol or paraformaldehyde. In contrast, PSA-FITC not only labelled the acrosome, but also the whole head and the flagellum. This aspect was especially marked after methanol fixation. The cytoplasmic droplet, when present, was also stained by PSA-FITC. Incubation with the calcium ionophore ionomycin induced a concentration and time-dependent increase in the number of acrosome reactions. Compared to spotted preparations, smear samples exhibited a high proportion of spermatozoa with damaged acrosome. In conclusion, PNA-FITC labelling was more accurate than PSA-FITC labelling to detect the acrosome of mouse spermatozoa. The fixation method (methanol vs. paraformaldehyde) had no influence on the staining pattern of PNA-FITC labelling, but spotted preparations are recommended to avoid mechanical damage to the acrosome. Ionophore challenge confirmed the existence of a calcium-dependent acrosome reaction in mouse spermatozoa and validated the use of PNA-FITC to quantify this physiological process. The present study illustrates important methodological considerations which need to be taken into account in order to design a reliable and reproducible protocol for the study of the acrosome reaction. PMID- 19554508 TI - Expression and distribution of GABAergic system in rat knee joint synovial membrane. AB - The GABAergic system, found in the adult mammalian brain and composed of gamma aminobutyric acid (GABA), GABA synthesizing enzyme glutamate decarboxylase (GAD) and GABA receptors, is also located in many peripheral nonneuronal tissues. Studies suggest that synovial membranes possess GABA, and that GABA participates in the control of the inflammatory response in rheumatoid arthritis (RA). However, no studies on the GABAergic system in synovial membranes have been done so far. Therefore, expression and distribution of the GABAergic system in the synovial membrane of the normal rat knee joint were investigated by reverse transcription-polymerase chain reaction (RT-PCR) analyses and immunohistochemistry. Results of RT-PCR analysis showed that mRNA encoding the GAD65 and GABAB receptor subunits necessary for the assembly of functional receptors, R1 and R2, are expressed in the synovial membrane. GAD and GABAB receptor subunits were localized in macrophage-like A cells of the synovial membrane. Macrophage-like A cells of the synovial membrane have a GABA production system and GABAB receptors, and GABA seems to play functional roles in the synovial membrane. PMID- 19554509 TI - Tumor cell expression of podoplanin correlates with nodal metastasis in esophageal squamous cell carcinoma. AB - Podoplanin is a mucin-like glycoprotein expressed in the lymphatic endothelium. It has been suggested to play a role in lymphangiogenesis, since podoplanin deficient mice were found to have dilated malfunctioning lymphatic vessels and lymphedema. High podoplanin expression in tumor cells was found to correlate with lymph node metastasis and poor clinical outcome in patients with oral squamous cell carcinoma (SCC). However, the prognostic significance of podoplanin expression in esophageal SCC remains unexplored. Herein, we studied podoplanin expression in 59 patients who underwent surgical resection of esophageal SCC, with 43 of them preceded by preoperative concurrent chemoradiotherapy (CCRT). We found that high podoplanin expression strongly correlated with clinical nodal metastasis (cN1; p=0.0063), which was associated with short survival (p=0.012). However, there was no direct association between high podoplanin expression and short survival. We also found that lymphatic vessel invasion in the resected esophagus was strongly associated with pathological nodal metastasis (pN1; p=0.00092). Our results suggest that podoplanin could also play a role in tumor aggressiveness in esophageal SCC, as well as in oral SCC. PMID- 19554510 TI - Efficient uptake of mannosylated proteins by a human Schwann cell line. AB - Complex carbohydrate structures are essential molecules of infectious microbes and host cells, and are involved in cell signaling associated with inflammatory and immune responses. The uptake of mannose-tailed glycans is usually carried out by macrophages, dendritic cells (DCs), and other professional phagocytes to trigger MHC class I- and MHC class II-restricted antigen presentation, and to promote T cell effector responses. Since Schwann cells (SCs) have been proposed as immunocompetent cells, we investigated whether a human cell line (ST88-14 cells) could bind mannosylated ligands in a specific manner. The saturation of uptake of mannosylated molecules by ST88-14 cells and the internalization and distribution pathway of these ligands were tested by cytometry and confocal plus electron microscopy, respectively. This uptake showed a dose-dependent increase, the saturation point being reached at high concentrations of mannosyl residues/240 mM mannose. Merging of man/BSA-FITC and S100 labeling showed their partial, but, significant colocalization. Ultrastructural analysis of ST88-14 cells after incubation with HRP-colloidal gold, without or with subsequent chasing at 37C, showed an initial location on the cell surface and temperature- and time-dependent internalization of the probe. Our findings suggest an efficient mannosylated ligand uptake system through putative lectin(s) that may be operational in inflammatory and immune responses. PMID- 19554511 TI - Aberrant CCND1 copies and cyclin D1 mRNA expression do not result in the production of functional cyclin D1 protein in anaplastic large cell lymphoma. AB - Scattered reports in the literature have shown that Cyclin D1 mRNA and protein may be expressed in anaplastic large cell lymphoma (ALCL). ALCLs are characterized by the presence of ALK translocations. Aberrant Cyclin D1 expression seems to promote proliferation in other types of lymphoma, while a growth promoting CCND1/TACSD1(TROP2) fusion product has also been described in tumors. Herein, we investigated 44 ALCL cases for chromosome 11 and CCND1 status (by FISH), cyclin D1 mRNA expression (by in situ hybridization and RT-PCR) and Cyclin D1 protein (by immunohistochemistry with two different monoclonal antibodies), as well as for the expression of Trop-2/GA733-1 (by immunohistochemistry). Polysomy of CCND1 (11q13) and chromosome 11 was found in 15/38 evaluated cases (39.5%). This change was specific for CD30+ neoplastic cells, as shown by double fluorescent staining. Neoplastic cells in the majority of ALCL expressed cyclin D1 mRNA (29/41 [70.7%]), in association with the presence of ALK translocations (p=0.024) and systemic, rather than cutaneous disease (p=0.021). Remarkably, however, Cyclin D1 protein was not detected in neoplastic cells (0/44 cases), neither were these found positive for Trop-2. In conclusion, aberrant copies of CCND1 / chromosome 11 may be observed in ALCL, probably as a consequence of the reported ploidy changes in these tumors. ALCL may often express cyclin D1 mRNA, which, however, does not result in the production of functional Cyclin D1 protein or Trop-2, suggesting that these proteins do not play a role in the pathogenesis of ALCL. PMID- 19554512 TI - Breast carcinoma vascularity: a comparison of manual microvessel count and Chalkley count. AB - Manual counting of microvessels as intratumoral microvessel density (MVD) and Chalkley counting have been used in several studies to assess the prognostic impact of vascularity in invasive breast carcinomas. In our present study, the aim was to evaluate the prognostic value of angiogenesis in invasive breast carcinoma assessed by MVD and Chalkley techniques in the same series of patients. A total of 498 breast carcinoma patients with median follow up time 85 months were evaluated. The tumour vascularity was quantified by both manual microvessel count (MVD) and Chalkley count in CD34 stained breast carcinoma slides by a single investigator blinded to clinical information. Other relevant clinicopathological parameters were noted, including breast cancer related death and both loco-regional and systemic relapse. The patients were stratified by converting MVD and Chalkley counts to categorical variables to assess prognostic impact, and results were compared. High vascular grades using MVD count did not demonstrate any prognostic significance for breast cancer specific survival (BCSS) or distant disease free survival (DDFS) either in whole patient group (BCSS, p=0.517, DDFS, p=0.301) or in non-treated node negative patients (p>0.05). Chalkley count showed prognostic significance for both DDFS and BCSS in whole patient group (p<0.001) and also in untreated node negative patient group (p<0.05). In multivariate analysis, Chalkley count, but not MVD, retained the prognostic value for BCSS (p=0.007) and DDFS (p=0.014). The Chalkley count for assessing angiogenesis in invasive breast carcinomas demonstrated prognostic value. The Chalkley method appears to be the better method in estimating the prognostic impact of vascularity in invasive breast carcinomas. PMID- 19554513 TI - In situ detection of APRIL-rich niches for plasma-cell survival and their contribution to B-cell lymphoma development. AB - A proliferation inducing ligand (APRIL) is one of the most recently cloned members of the tumor necrosis factor (TNF) family. Early experiments implicated a pathophysiological role for APRIL in the promotion of solid tumors. Later, identification of APRIL receptors on B lymphocytes indicated a physiological role for APRIL in humoral responses. We have been able to generate antibodies that detect APRIL protein in human tissues. The study of in situ APRIL expression showed that APRIL mainly regulates late stages of B-cell humoral responses. It also provided evidence that APRIL may modulate tumor development in patients, but only for specific B-cell malignancies. Here, we will review to what extent fine characterization of in situ expression adds valuable information on APRIL (patho) physiological functions. PMID- 19554514 TI - Genetic mouse models for the functional analysis of the perifibrillar components collagen IX, COMP and matrilin-3: Implications for growth cartilage differentiation and endochondral ossification. AB - The mutual interaction of the two supramolecular compartments, the fibrillar and extrafibrillar matrix is a prerequisite for stability and integrity of the cartilage extracellular matrix. The fibrillar periphery, composed of collagen IX, matrilins and cartilage oligomeric matrix protein (COMP) among other components, constitutes the interface which mediates interactions between the two compartments. Mutations in these peripheral macromolecules cause a broad spectrum of skeletal conditions such as pseudo-achondroplasia (PSACH) and multiple epiphyseal dysplasia (MED), which severely affect the organization and integrity of the cartilage growth matrix in humans. Transgenic and knockout mouse models for collagen IX, matrilin-3 and COMP and combinations thereof display cartilage abnormalities and pathologies of varying severity. Absence of collagen IX appears to cause the most severe growth plate phenotype with a profoundly disturbed morphological organization affecting size and shape of the long bones. Notably, similar growth plate phenotypes, including irregularities in the proteoglycan content, hypocellular central regions, disorganized proliferation columns with atypically shaped and oriented chondrocytes and alterations in the hypertrophic zone are observed in transgenic mice lacking other macromolecules or carrying mutations therein. These include collagens II and XI, integrin subunits, integrin linked kinase (ILK), HIF-1alpha, VEGFalpha and the tumor suppressor PTEN. Notably, mutations in ciliar proteins such as Kif3alpha, polaris or Smo/Gli severely affect the ability of chondrocytes to move and to become arranged in columns. Absence or mutational changes of a variety of different, non-related cartilage macromolecules apparently cause similar pathologies and abnormalities of the growth cartilage, suggesting a limited number of underlying molecular mechanisms. PMID- 19554516 TI - Revolutionary impact of PET and PET-CT on the day-to-day practice of medicine and its great potential for improving future health care. AB - In this communication, we present an overview of the impact and advantages of PET and PET-CT fusion imaging in the practice of medicine. We also discuss the evolution of this promising molecular imaging technique since its inception and the future prospects of the combined structure-function approach. Superior contrast resolution, accurate quantification and above all optimal image quality aid in improved diagnosis of many serious disorders including cancer. We speculate that this powerful imaging approach will almost completely replace most other conventional methods in the future. Currently, 18[F]-fluorode- -oxyglucose (FDG) is the main radiopharmaceutical employed for PET studies around the globe. With the availability of high quality PET images on a routine basis in most centres around the world and the likelihood that several other useful PET tracers will be approved in the near future for routine clinical applications, this technique will likely become essential in almost any medical disorder. PMID- 19554515 TI - Cytoplasmic inclusions of TDP-43 in neurodegenerative diseases: a potential role for caspases. AB - TAR DNA-binding protein-43 (TDP-43) proteinopathies are classified based upon the extent of modified TDP-43 inclusions and include a growing number of neurodegenerative diseases including amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration with ubiquitin immunoreactive, tau negative inclusions (FTLD-U) and FTLD with motor neuron disease (FTLD-MND). In addition, TDP-43 inclusions have also been identified in a number of other neurodegenerative disorders including Alzheimer's disease, corticobasal degeneration, Lewy body related diseases and Pick's disease. Current understanding suggests that in these diseases, TDP-43 is relocated from the nucleus to the cytoplasm and sequestered into inclusions that contain modified TDP-43. Major modifications of TDP-43 have been identified as being hyperphosphorylation and proteolytic cleavage by caspases. In this review a summary of the major findings regarding the proteolytic modification of TDP-43 will be discussed as well as potential toxic-gain mechanisms these fragments may cause including cytoskeletal disruptions. PMID- 19554517 TI - Assessment of accuracy and precision of (99m)Tc-HEPIDA clearances determined by means of a simplified method. AB - BACKGROUND: The aim of the present study was the assessment of the accuracy and precision of our own simplified method for the determination of (99m)Tc-HEPIDA liver clearance. MATERIAL AND METHODS: It has been assumed that archived results of plasma clearance (Cl(Pl)) and hepatic (Cl(Hp)), determined by means of multisample methods, could be legitimately used as a reference standard. The accuracy and precision of the simplified method was assessed by means of a Monte Carlo method alternatively utilizing three blood sampling times (T) of 68, 75 and 83 minutes post i.v. administration of (99m)Tc-HEPIDA. The corresponding alternative three urine voiding times (Y) were: 75, 80, and 95 min p.i. The analysed model was created accepting values of Cl(Pl) and Cl(Hp), of administered activity A(p) and parameters of biexponential function, describing the concentration C(t) decrease of the radiopharmaceutical (RF) in plasma during time as real values. Using the function C(t) for each individual, the plasma concentrations of RF at three sampling times, urinary clearance (Cl(Pl) - Cl(Hp)), and voided activity (A(Ur)(Y)) were calculated. Simulated random errors were added to the assumed blood sampling times T and to voiding time Y. To the activity A(p) and A(Ur)(Y), and RF plasma concentrations random errors were added, assuming normal distribution with relative SD from 0 to 5% and then clearance values were computed. For each process there were 5000 repeated simulated determinations. The accuracy of the simplified methods was assessed by comparing mean values of simulated clearance computations with the reference. Comparison of standard deviations with mean uncertainties enabled us to gain insight into the degree of agreement of the estimator of relative uncertainty with the coefficient of variation as a measure of precision. RESULTS: There were strong correlations between the reference clearance values and the mean values of determinations by means of the simplified procedure (r > 0.93). The correlations were practically insensitive to the uncertainty of pipetting. The lines of regression differed slightly from the lines of identity, giving an indication that there was a systematic error involved; it amounted to +4 ml/min at Cl(Pl) = 60 ml/min and to -7 ml/min for Cl(Pl) of 370 ml/min. For Cl(Pl) a bias of +6 ml/min was found for a clearance value of 16 ml/min and -13 ml/min at Cl(Pl) > 300 ml/min. At uncertainty of pipetting of 2%, a precision of 6-7% was found for Cl(Pl) of 300 ml/min. For Cl(Pl) of 200 and 150 ml/min the corresponding precisions were 7-8% and 10%, respectively. For Cl(Pl) of 200, 150 and 100 ml/min the corresponding precisions were 10, 12 and 17%, respectively. These precisions are 5 percent worse than those that were obtained from determinations by means of multisampling procedures. PMID- 19554520 TI - [Letter from the secretary of the Italian Society of Nephrology]. PMID- 19554518 TI - Radioiodine - an attractive alternative to surgery in large non-toxic multinodular goitres. AB - Sporadic non-toxic goitre (SNG) is defined as thyroid enlargement in a euthyroid patient living in an area without endemic goitre. Because thyroid hormone function is within the reference range, the main problems are the signs of thyroid enlargement, thus reducing the size of the goitre is undoubtedly the primary goal of therapy. Although SNG is a common disease, there is no single optimal management of treatment strategy. L-thyroxine treatment, although only moderately effective and with reported reductions in goitre volume of less than 30%, is still used. Moreover, in patients over 60 years of age, long-term TSH suppression treatment may cause iatrogenic hyperthyroidism and is associated with a significant risk of paroxysmal atrial fibrillation as well as osteoporosis, especially in postmenopausal women. In patients with huge goitres, the surgical removal of a gland is made most frequently. The great advantage of thyroid surgery is its immediate effect. Unfortunately, surgery carries a risk of goitre recurrence and complications, both surgical and anaesthesia-related. For those patients who do not want to be operated on, or have contraindications for invasive treatment, and taking into consideration the low efficacy of L-thyroxine treatment, the best option is radioiodine. Despite encouraging reports of the high efficacy of radioiodine in the treatment of SNG this method is still not commonly used by many clinicians. In our work we would like to point out the efficacy, adverse effects, and contraindications of using iodine-131. Going through the advantages and disadvantages of all accessible methods of treatment of SNG, we would like to focus on using radioiodine as an attractive alternative to surgery. PMID- 19554521 TI - [FGF-23: a new player in CKD-MBD]. PMID- 19554522 TI - [Cosmc: the ''chaperone'' toward the understanding of IgA Nephropathy?]. PMID- 19554523 TI - [The dark side of the (half) moon]. PMID- 19554524 TI - [Pulse high-volume hemofiltration (p-HVHF) in sepsis: extracorporeal treatment for renal function replacement and immune dysregulation]. PMID- 19554525 TI - [To immunosuppress or to tolerate: that is the question!]. PMID- 19554526 TI - [''The kidney, development, repair and regeneration''. News from the KIDSTEM International Conference, Liverpool, 17-19 September 2008]. PMID- 19554527 TI - [Nephroangiosclerosis and ischemic nephropathy: two different entities or two renal manifestations of the same systemic cardiovascular disease?]. AB - Nephroangiosclerosis (NAS) is increasingly diagnosed in adult and elderly patients with slowly progressive chronic renal insufficiency. Since these patients usually present with arterial hypertension, this is considered the main cause of NAS (sometimes called, in fact, hypertensive NAS or hypertensive nephropathy). However, there is evidence that other factors such as aging, black race, smoking, and metabolic disturbances contribute to the development and progression of the disease. In some patients, these factors may be prominent while hypertension may be mild or even absent: this form has been denominated ischemic nephropathy (IN). Are NAS and IN really two different diseases or just different presentations of cardiovascular disease involving the kidney? The latter hypothesis is supported by evidence that (a) NAS and IN share a relative aspecificity in their clinical symptoms (low proteinuria, microhematuria, high blood pressure, dyslipidemia) and histopathological features (as determined in the few cases that undergo a kidney biopsy), and (b) there is a high likelihood that atheromatous and hypertensive lesions coexist in the same patient. In this ''Controversy in Nephrology'', Rosario Cianci and Alessandro Zuccala' analyze this issue and try to answer the following questions: 1 - Are NAS and IN two different diseases or two different expressions of the same disease? Rosario Cianci, ''They are two different diseases''. Alessandro Zuccala', ''They represent two different expressions of the same disease''. 2 - Is the pathogenesis different in nephroangiosclerosis and IN? Rosario Cianci, ''The pathogenesis is high blood pressure in NAS and renal ischemia in IN''. Alessandro Zuccala', ''NAS and IN share the same multifactorial pathogenesis: vascular metabolic alterations can cause chronic renal ischemia with or without hypertension''. 3 - Is a biopsy necessary for the diagnosis? Rosario Cianci, ''Yes, it is''. Alessandro Zuccala', ''No, it is not''. 4 - Is it possible to prevent or to slow the progression of the renal damage in this (these) disease(s)? Rosario Cianci, ''Yes it is, by reducing blood pressure''. Alessandro Zuccala', ''Normalization of blood pressure is not enough but all the other risk factors of vascular damage must be addressed, when possible''. PMID- 19554528 TI - [Rationale, pros and cons of GFR estimation: the Cockcroft-Gault and MDRD equations]. AB - Current guidelines suggest to evaluate kidney function by estimates of the glomerular filtration rate (GFR) calculated using the Cockcroft-Gault equation or the simplified equation of the Modification of Diet in Renal Disease (MDRD) study. Cockcroft and Gault developed a method to predict an individual's creatinine clearance without urinary data but on the basis of average creatinine clearance values corrected for age and body weight. The equation needs information about gender, age, weight, and serum creatinine. The precision of the equation is biased by overweight because the equation is based on the assumption that creatinine clearance linearly reflects body weight. The MDRD equation derives from a multiple regression used to analyze the relation of GFR measurements by a radioisotope technique over serum creatinine after data linearization by logarithm transformation and with control for gender, age, ethnicity and other variables. The equation has not been validated for GFR >60 mL/min x 1.73 m2 because the study did not include healthy persons. The two equations often give conflicting estimates of GFR. Nephrologists have to understand the rationale of the two equations for the correct interpretation of these discrepancies. PMID- 19554529 TI - [Treatment of HCV-associated cryoglobulinemic glomerulonephritis]. AB - HCV-related membranoproliferative glomerulonephritis is the most common cause of hepatitis C-associated renal disease. Its treatment is still under debate and based on scant experimental evidence. The recommended therapeutic strategy depends on the severity of the kidney disease. The first-line treatment for patients with mild to moderate clinical and histological kidney damage is antiviral therapy with pegylated interferon alpha and ribavirin for 48 weeks combined with symptomatic treatment (diuretics, angiotensin converting enzyme inhibitors and angiotensin receptor blockers). In case of severe renal involvement (nephrotic syndrome, nephritic syndrome and/or progressive renal failure, high activity score of glomerulonephritis on light microscopy), the initial treatment may consist of sequential administration of immunosuppressive therapies (plasmapheresis, corticosteroids and cyclophosphamide) and antiviral agents, although no definitive data are yet available from the literature. B-cell depleting agents such as rituximab may be an alternative to conventional therapy in refractory or intolerant patients. Large randomized and controlled clinical trials are needed to establish guidelines for the treatment of HCV-related cryoglobulinemic glomerulonephritis. PMID- 19554530 TI - [Resistant hypertension in non-dialysis chronic kidney disease]. AB - Resistant hypertension is defined as blood pressure that remains above the target of <140/90 mm Hg in the general population and <130/80 mm Hg in people with diabetes mellitus or chronic kidney disease (CKD) in spite of the use of at least three full-dose antihypertensive drugs including a diuretic, or as blood pressure that reaches the target by means of four or more drugs. Hypertension is a frequent complication in CKD and a determining factor in the progression of renal damage, especially in proteinuric and diabetic patients, as well as contributing to a high cardiovascular risk. Clinical practice guidelines recommend blood pressure levels below 130/80 mm Hg in all CKD patients, but the target is reached in only a small proportion (10-20%), both in nephrology and non-nephrology settings. The resistance to antihypertensive treatment may be considered one of the causes of the poor achievement of blood pressure targets in CKD patients. PMID- 19554531 TI - [C-reactive protein in the assessment of iron status in patients on hemodialysis]. AB - Iron availability is a prerequisite for an efficient hematopoietic response to erythropoietin. Dynamic evaluation of iron status is difficult in hemodialysis patients and can be further complicated by the presence of an inflammatory state. Several cytokines, in particular interleukin 6 (IL-6), stimulate the production of hepcidin in the liver. This hormone is the main regulator of the extracellular iron concentration through its effect on the iron channel ferroportin, present in several cell types. IL-6 is also the major stimulus for the production of C reactive protein (CRP), a nonspecific but sensitive marker of inflammation. Measurement of hepcidin is technically difficult and has so far been limited to research. On the other hand, measurement of CRP, which is both sensitive and easily measurable with automated techniques, might possibly be used as a surrogate measure of iron status in hemodialysis patients. Several studies have suggested the value of CRP in this context, but they dealt with small patient groups and single-time-point measurements. Even the definition of normal values of CRP in dialysis patients is uncertain. During the period between 2003 and 2007, we performed 8322 measurements of CRP in 401 hemodialysis patients followed for 3-60 months. All parameters of iron balance (serum iron, TSAT, percent hypochromic RBC and Hgb concentration in reticulocytes) were clearly affected by the presence of an inflammatory state. We believe that measurement of CRP must be part of the routine hematological assessment of hemodialyzed patients to allow the correct interpretation of data in anemia treatment. PMID- 19554532 TI - [Aldosterone and kidney damage: clinical implications]. AB - In addition to data regarding its effects on the heart, brain and blood vessels, extensive evidence has emerged about the contribution of aldosterone to kidney damage. This has mainly been studied in the setting of experimental models of salt-sensitive hypertension but has been confirmed also in other animal models. The evidence is supported by a clear causal relationship between aldosterone infusion and development of kidney damage and its reversal after aldosterone blockade. Since 2001, clinical data have been obtained on the antiproteinuric effect of aldosterone antagonists added to ACE inhibitors or angiotensin II receptor blockers. Unfortunately, the long-term findings are still scanty, except for those obtained in two one-year studies. Altogether, this therapeutic approach appears relatively safe and effective; however, larger studies on patients with a wider range of chronic kidney disease severities and longer follow-up are needed to confirm it. PMID- 19554533 TI - [Cyclers for APD: current potential and possible developments]. AB - The use of automated peritoneal dialysis (APD) is rapidly increasing worldwide. The success of this treatment is closely linked to the development of new mechanical devices (cyclers). At present, the cyclers available in Italy are the Baxter Home Choice Pro, the Fresenius Sleep Safe, and the Gambro Serena. Today's cyclers are characterized by a user-friendly, intuitive interface and sophisticated software. The miniaturization of components has led to a significant reduction of the weight and size of these machines and great flexibility in treatment scheduling. The memory card (patient card) and the possibility of remote control (teledialysis) allow continuous monitoring of the patients and their compliance to treatment. The cyclers of the future could optimize their performance by biosensors able to assess, in real time, the dialysis dose while sensors placed on the drainage line could facilitate the early diagnosis of peritonitis. The peritoneal clearance could be increased with the development of continuous-flow peritoneal dialysis (CFPD). Finally, the introduction of cyclers able to prepare individually tailored solutions different from the standard ones could better meet the clinical needs of the patients and reduce the exposure of the peritoneal membrane to high concentrations of glucose. PMID- 19554534 TI - [Midarm arteriovenous fistula: single-center experience]. AB - In uremic patients on hemodialysis, a first vascular access using native vessels tailored into a radiocephalic arteriovenous fistula (AVF) on the wrist is the gold standard in vascular access quality. However, among the uremic population the percentage of older patients affected by diabetes or by severe generalized vascular disease is growing. In these patients distal radiocephalic AVFs often have a limited survival. This induced us to design a technique for tailoring vascular accesses at a proximal site. In the past two years (2006-2007) we created 19 AVFs at the midarm position (midarm AVF) using the proximal section of the radial artery. Midarm AVF was the second choice in 70% of patients after failure of a distal AVF and the first choice in 30% of patients. The survival of this type of access has been excellent. We have registered only one thrombosis after 6 months. In four cases access was interrupted because of the death of the patients. The remaining 14 accesses are working perfectly well. Our experience leads us to conclude that midarm AVF is not only an important intermediate step after the failure of a distal AVF and before the application of a proximal AVF, but in a population of patients with diabetes and generalized vascular disease it should probably be proposed as the AVF of choice. PMID- 19554535 TI - [Multiple peritrochanteric and pubic calcifications in a young woman on hemodialysis with severe renal osteodystrophy successfully treated with sevelamer+cinacalcet+paracalcitol combination therapy]. AB - Secondary hyperparathyroidism is a frequent complication of chronic renal failure that can induce severe bone disease and negatively influence the cardiovascular outcome. Therefore, nephrologists should attempt to reach the targets recommended by national and international guidelines using all the available therapeutic strategies. We describe the case of a 37-year-old woman affected by spina bifida and myelomeningocele who had been on hemodialysis since 1993. In July 2006 she developed secondary hyperparathyroidism complicated by peritrochanteric calcifications which did not respond to standard therapy. Because it was impossible to perform a parathyroidectomy, we started medical therapy with a combination of sevelamer hydrochloride, paracalcitol and cinacalcet, which resulted in progressive improvement of laboratory data and osteodystrophy. A diagnosis of mixed secondarytertiary hyperparathyroidism was made, but a progressive increase in iPTH to very high levels suggested a rapid evolution toward a pure tertiary form. PMID- 19554536 TI - [The difficult start of nephrology in Rome]. AB - Nephrology in Rome began in the 1960s with the arrival of Ernico Fiaschi in the wake of Cataldo Cassano at the Institute of Medical Pathology (later on Clinica Medica II). A group of doctors interested in nephrology was set up, with among them Giulio A. Cinotti, who was to become full professor of nephrology at the University of Rome ''La Sapienza'' in 1980. By the end of the 1960s, the renal transplant activity had become an important asset at the Institute of Surgical Pathology (later on Clinica Chirurgica II) thanks to Paride Stefanini. A chair of surgical nephrology was instituted at the Urology Clinics of Ulrico Bracci; the chair was first held by Nicola Cerulli, who developed an intensive hemodialysis program. Around the same time, the Center for the Research and Treatment of Arterial Hypertension and Kidney Diseases became operational at the hospitals of Rome (under the responsibility of Vito Cagli at the Policlinico Umberto I), while a nephrology and dialysis unit, directed by Giancarlo Ruggieri, was set up at the San Giacomo Hospital. Many nephrology-related ''cultural'' activities started to be undertaken thanks to the ''Gruppo Laziale di Nefrologia Medica e Chirurgica'' founded by Drs Cagli, Cerulli, and Cinotti. Two national congresses were organized by Giulio Cinotti in 1979 (Fiuggi) and 1992 (Rome). PMID- 19554538 TI - [Full colour dialysis]. PMID- 19554537 TI - [Clinical risk management in the context of JCI certification]. PMID- 19554539 TI - Mesenchymal origin of hepatic stellate cells, submesothelial cells, and perivascular mesenchymal cells during mouse liver development. PMID- 19554540 TI - Hepatic recruitment of the inflammatory Gr1+ monocyte subset upon liver injury promotes hepatic fibrosis. AB - In addition to liver-resident Kupffer cells, infiltrating immune cells have recently been linked to the development of liver fibrosis. Blood monocytes are circulating precursors of tissue macrophages and can be divided into two functionally distinct subpopulations in mice: Gr1(hi) (Ly6C(hi)) and Gr1(lo) (Ly6C(lo)) monocytes. The role of these monocyte subsets in hepatic fibrosis and the mechanisms of their differential recruitment into the injured liver are unknown. We therefore characterized subpopulations of infiltrating monocytes in acute and chronic carbon tetrachloride (CCl(4))-induced liver injury in mice using flow cytometry and immunohistochemistry. Inflammatory Gr1(hi) but not Gr1(lo) monocytes are massively recruited into the liver upon toxic injury constituting an up to 10-fold increase in CD11b(+)F4/80(+) intrahepatic macrophages. Comparing wild-type with C-C chemokine receptor (CCR2)-deficient and CCR2/CCR6-deficient mice revealed that CCR2 critically controls intrahepatic Gr1(hi) monocyte accumulation by mediating their egress from bone marrow. During chronic liver damage, intrahepatic CD11b(+)F4/80(+)Gr1(+) monocyte-derived cells differentiate preferentially into inducible nitric oxide synthase-producing macrophages exerting proinflammatory and profibrogenic actions, such as promoting hepatic stellate cell (HSC) activation, T helper 1-T cell differentiation and transforming growth factor beta (TGF-beta) release. Impaired monocyte subset recruitment in Ccr2(-/-) and Ccr2(-/-)Ccr6(-/-) mice results in reduced HSC activation and diminished liver fibrosis. Moreover, adoptively transferred Gr1(hi) monocytes traffic into the injured liver and promote fibrosis progression in wild-type and Ccr2(-/-)Ccr6(-/-) mice, which are otherwise protected from hepatic fibrosis. Intrahepatic CD11b(+)F4/80(+)Gr1(+) monocyte-derived macrophages purified from CCl(4)-treated animals, but not naive bone marrow monocytes or control lymphocytes, directly activate HSCs in a TGF-beta-dependent manner in vitro. CONCLUSION: Inflammatory Gr1(+) monocytes, recruited into the injured liver via CCR2-dependent bone marrow egress, promote the progression of liver fibrosis. Thus, they may represent an interesting novel target for antifibrotic strategies. PMID- 19554541 TI - HFE C282Y/H63D compound heterozygotes are at low risk of hemochromatosis-related morbidity. AB - The risk of hemochromatosis-related morbidity is unknown among HFE compound heterozygotes (C282Y/H63D). We used a prospective population-based cohort study to estimate the prevalence of elevated iron indices and hemochromatosis-related morbidity for compound heterozygotes. In all, 31,192 subjects of northern European descent were genotyped for HFE C282Y and H63D. An HFE-genotype stratified random sample of 1,438 subjects, followed for an average of 12 years to a mean age of 65 years, completed questionnaires and gave blood. Clinical examinations were blinded to HFE genotype. A total of 180 (84 males) clinically examined C282Y/H63D participants were compared with 330 (149 males) controls with neither HFE mutation; 132 (65 males) and 270 (122 males), respectively, had serum iron measures at both timepoints. Mean serum ferritin (SF) and transferrin saturation (TS) were significantly greater for male and female compound heterozygotes than for wild-types at baseline and follow-up (all P < 0.02) except for females who were premenopausal at baseline, where SF was similar in both genotype groups. For subjects with serum measures from both baseline and follow up, mean SF and TS levels did not change significantly for men or for postmenopausal women, but for premenopausal women SF levels increased from 43 to 109 microg/L for compound heterozygotes and from 35 to 64 microg/L for wild-types (both P < 0.001). Male and female compound heterozygotes had a similar prevalence of hemochromatosis-related morbidity to wild-types. One of 82 males and zero of 95 females had documented iron overload-related disease. CONCLUSION: For male compound heterozygotes, mean iron indices do not change during middle age but for female compound heterozygotes menopause results in increased mean SF. Although compound heterozygotes might maintain elevated iron indices during middle age, documented iron overload-related disease is rare. PMID- 19554542 TI - L-ornithine phenylacetate attenuates increased arterial and extracellular brain ammonia and prevents intracranial hypertension in pigs with acute liver failure. AB - Hyperammonemia is a feature of acute liver failure (ALF), which is associated with increased intracranial pressure (ICP) and brain herniation. We hypothesized that a combination of L-ornithine and phenylacetate (OP) would synergistically reduce toxic levels of ammonia by (1) L-ornithine increasing glutamine production (ammonia removal) through muscle glutamine synthetase and (2) phenylacetate conjugating with the ornithine-derived glutamine to form phenylacetylglutamine, which is excreted into the urine. The aims of this study were to determine the effect of OP on arterial and extracellular brain ammonia concentrations as well as ICP in pigs with ALF (induced by liver devascularization). ALF pigs were treated with OP (L-ornithine 0.07 g/kg/hour intravenously; phenylbutyrate, prodrug for phenylacetate; 0.05 g/kg/hour intraduodenally) for 8 hours following ALF induction. ICP was monitored throughout, and arterial and extracellular brain ammonia were measured along with phenylacetylglutamine in the urine. Compared with ALF + saline pigs, treatment with OP significantly attenuated concentrations of arterial ammonia (589.6 +/- 56.7 versus 365.2 +/- 60.4 mumol/L [mean +/- SEM], P= 0.002) and extracellular brain ammonia (P= 0.01). The ALF-induced increase in ICP was prevented in ALF + OP-treated pigs (18.3 +/- 1.3 mmHg in ALF + saline versus 10.3 +/- 1.1 mmHg in ALF + OP-treated pigs;P= 0.001). The value of ICP significantly correlated with the concentration of extracellular brain ammonia (r(2) = 0.36,P< 0.001). Urine phenylacetylglutamine levels increased to 4.9 +/- 0.6 micromol/L in ALF + OP-treated pigs versus 0.5 +/- 0.04 micromol/L in ALF + saline-treated pigs (P< 0.001). CONCLUSION: L-Ornithine and phenylacetate act synergistically to successfully attenuate increases in arterial ammonia, which is accompanied by a significant decrease in extracellular brain ammonia and prevention of intracranial hypertension in pigs with ALF. PMID- 19554543 TI - Primary biliary cirrhosis. PMID- 19554545 TI - Management of patients with hepatitis C virus genotype 2 or 3: comments on updated American Association for the Study of Liver Diseases practice guidelines. PMID- 19554546 TI - Comment on the updated AASLD practice guidelines for the diagnosis, management, and treatment of hepatitis C: treating active drug users. PMID- 19554547 TI - Image of the month. Albinism, cyclic neutropenia, and ceroid pigment in the liver. PMID- 19554550 TI - Things that go BMP in the liver: bone morphogenetic protein 6 and the control of body iron homeostasis. PMID- 19554551 TI - Solid organ transplantation in HIV-infected individuals: an update. AB - In the era of highly active antiretroviral therapy (HAART), the survival of patients with HIV has improved. Increasing morbidity and mortality are now related to chronic liver and kidney disease. Transplantation in HIV patients has been reported for nearly two decades and outcomes have generally improved in the HAART era. This review summarises the published experiences with liver and kidney transplantation in HIV patients. PMID- 19554552 TI - An age-adapted approach for the use of D-dimers in the exclusion of deep venous thrombosis. AB - A normal D-dimer (DD) concentration for the exclusion of deep venous thrombosis (DVT) has a low specificity in older patients and compression ultrasonography is often required. Three D-dimer assays, STA Liatest, Tina-quant, and Innovance, are evaluated in symptomatic outpatients suspected for DVT with emphasis on its performance in older patients by using different cut-off levels. This study includes 466 outpatients suspected for having DVT. The diagnostic accuracy, measured as sensitivity and area under the curve of the receiver operation characteristic curve is good for all DD assays. The specificity of the DD assays combined with a low pretest probability varies from 42.6 to 51.5%. The specificity of the three DD assays in patients > or = 60 years varies, however, between 24.6 and 40.9%. Several cut-off values in different age-subgroups are studied. For patients < 60 years, the most accurate cut-off value is 500 microg/L for all DD assays. For patients > or = 60 years, a threshold of 750 microg/L has the best results with NPV of 100% for all assays and specificity of 48.5% (STA Liatest), 60.6% (Tina-quant), and 49.2% (Innovance), respectively. For the three assays, the number needed to test (NNT) decreases in both subgroups of patients compared to the standard algorithm. A cut-off level of 750 microg/L for patients > or = 60 years improves the clinical performance of DD assays in combination with the PTP score without the loss of NPV. The NNT improves substantially with an age-adapted algorithm. PMID- 19554554 TI - A combinatorial study of inverse Heusler alloys by first-principles computational methods. AB - In continuation of our recent combinatorial work on 810 X(2)YZ full Heusler alloys, a computational study of the same class of materials but with the inverse (XY)XZ crystal structure has been performed on the basis of first-principles (GGA) total-energy calculations using pseudopotentials and plane waves. The predicted enthalpies of formation evidence 27 phases to be thermochemically stable against the elements and the regular X(2)YZ type. A chemical-bonding study yields an inherent tendency for structural distortion in a majority of these alloys, and we predict the existence of the new tetragonal phase Fe(2)CuGa (P4(2)/ncm; a = 5.072 A, c = 7.634 A; c/a approximately 1.51) with a saturation moment of mu = 4.69 micro(B) per formula unit. Thirteen more likewise new, isotypical phases are predicted to show essentially the same behavior. Six phases turn out to be the most stable in the inverse tetragonal arrangement. The course of the magnetic properties as a function of the valence-electron concentration is analyzed using a Slater-Pauling approach. PMID- 19554553 TI - Overexpression of nucleolin in engrafted acute myelogenous leukemia cells. PMID- 19554555 TI - Aten--an application for the creation, editing, and visualization of coordinates for glasses, liquids, crystals, and molecules. AB - Aten is a tool for Linux, Mac, and Windows platforms to ease the creation, editing, and visualization of coordinates for use in, for example, molecular dynamics simulations. The code handles gas-phase molecules in addition to crystals, surfaces, and liquids, providing standard tools to edit "by the atom" along with specific "by the box" methods suited to periodic systems, including full crystallographic spacegroup packing definitions. Visualization of systems encompasses the standard drawing styles and may be mixed with an arbitrary number of other basic objects, such as arrows and geometric objects allowing creation of scenes involving vector fields, coarse-grained particles, etc. Standard molecular mechanics forcefields can be read in, edited, applied to molecules using the built-in chemical typing language, and subsequently used to calculate energies and forces, minimize energies with respect to coordinates, decompose energies into contribution by molecule type, etc. Monte Carlo methods are available with which to generate random configurations of N-component systems, or solvate around molecules or in specific regions within existing configurations. Supported file formats are governed by user-defined "filters," which provide flexibility of input/output as well as the ability to define custom or extended formats of traditional files. Forcefield descriptions for loaded systems can be output and formatted for use as input to common codes. All features can be accessed by a comprehensive GUI and scripting language based on the C syntax. PMID- 19554556 TI - How hot? Systematic convergence of the replica exchange method using multiple reservoirs. AB - We have devised a systematic approach to converge a replica exchange molecular dynamics simulation by dividing the full temperature range into a series of higher temperature reservoirs and a finite number of lower temperature subreplicas. A defined highest temperature reservoir of equilibrium conformations is used to help converge a lower but still hot temperature subreplica, which in turn serves as the high-temperature reservoir for the next set of lower temperature subreplicas. The process is continued until an optimal temperature reservoir is reached to converge the simulation at the target temperature. This gradual convergence of subreplicas allows for better and faster convergence at the temperature of interest and all intermediate temperatures for thermodynamic analysis, as well as optimizing the use of multiple processors. We illustrate the overall effectiveness of our multiple reservoir replica exchange strategy by comparing sampling and computational efficiency with respect to replica exchange, as well as comparing methods when converging the structural ensemble of the disordered Abeta(21-30) peptide simulated with explicit water by comparing calculated Rotating Overhauser Effect Spectroscopy intensities to experimentally measured values. PMID- 19554557 TI - Computational determination of fundamental pathway and activation barriers for acetohydroxyacid synthase-catalyzed condensation reactions of alpha-keto acids. AB - Acetohydroxyacid synthase (AHAS) is the first common enzyme in the biosynthetic pathway leading to the production of various branched-chain amino acids. AHAS is recognized as a promising target for new antituberculosis drugs, antibacterial drugs, and herbicides. Extensive first-principles quantum mechanical (QM) and hybrid quantum mechanical/molecular mechanical (QM/MM) calculations have enabled us, in this study, to uncover the fundamental reaction pathway, determine the activation barriers, and obtain valuable insights concerning the specific roles of key amino acid residues for the common steps of AHAS-catalyzed condensation reactions of alpha-keto acids. The computational results reveal that the rate determining step of the AHAS-catalyzed reactions is the second reaction step and that the most important amino acid residues involved in the catalysis include Glu144', Gln207', Gly121', and Gly511 that form favorable hydrogen bonds with the reaction center (consisting of atoms from the substrate and cofactor) during the reaction process. In addition, Glu144' also accepts a proton from cofactor thiamin diphosphate (ThDP) through hydrogen bonding during the catalytic reaction. The favorable interactions between the reaction center and protein environment remarkably stabilize the transition state and, thus, lower the activation barrier for the rate-determining reaction step by approximately 20 kcal/mol. The activation barrier calculated for the rate-determining step is in good agreement with the experimental activation barrier. The detailed structural and mechanistic insights should be valuable for rational design of novel, potent AHAS inhibitors that may be used as promising new anti-tuberculosis drugs, antibacterial drugs, and/or herbicides to overcome drug resistance problem. PMID- 19554559 TI - The effect of presentation paradigm on syntactic processing: An event-related fMRI study. AB - An event-related fMRI study was conducted to investigate the effect of two different sentence presentation paradigms-rapid serial visual presentation (RSVP) and whole sentence presentation-on syntactic processing. During scanning, sentences were presented using one of the two presentation paradigms and were followed by a short delay and a probe to verify sentence comprehension. The delay was included in an attempt to separate sentence-related activity from probe related activity. The behavioral data showed a main effect of syntactic complexity for reaction time and accuracy, and accuracy revealed an interaction between complexity and the presentation paradigm employed-RSVP produced many more errors for syntactically complex sentences than did whole sentence presentation. The imaging data revealed a syntactic complexity effect during the sentence phase in left BA 44 and during the probe phase in left BA 44 and the left posterior MTG. In addition, timecourse analysis revealed that these two regions also showed an interaction between complexity and presentation paradigm such that there was no complexity effect during RSVP but a significant effect during whole sentence presentation. In addition to finding that these two presentation paradigms differentially affected syntactic processing, there were main effects within the visual pathway (V1/V2 vs. V5) and the hippocampus that revealed significant differences in activation between the paradigms. PMID- 19554558 TI - Auditory orienting and inhibition of return in mild traumatic brain injury: a FMRI study. AB - The semiacute phase of mild traumatic brain injury (mTBI) is associated with deficits in the cognitive domains of attention, memory, and executive function, which previous work suggests may be related to a specific deficit in disengaging attentional focus. However, to date, there have only been a few studies that have employed dynamic imaging techniques to investigate the potential neurological basis of these cognitive deficits during the semiacute stage of injury. Therefore, event-related functional magnetic resonance imaging was used to investigate the neurological correlates of attentional dysfunction in a clinically homogeneous sample of 16 patients with mTBI during the semiacute phase of injury (<3 weeks). Behaviorally, patients with mTBI exhibited deficits in disengaging and reorienting auditory attention following invalid cues as well as a failure to inhibit attentional allocation to a cued spatial location compared to a group of matched controls. Accordingly, patients with mTBI also exhibited hypoactivation within thalamus, striatum, midbrain nuclei, and cerebellum across all trials as well as hypoactivation in the right posterior parietal cortex, presupplementary motor area, bilateral frontal eye fields, and right ventrolateral prefrontal cortex during attentional disengagement. Finally, the hemodynamic response within several regions of the attentional network predicted response times better for controls than for patients with mTBI. These objective neurological findings represent a potential biomarker for the behavioral deficits in spatial attention that characterize the initial recovery phase of mTBI. PMID- 19554560 TI - Distributed cell assemblies for general lexical and category-specific semantic processing as revealed by fMRI cluster analysis. AB - Here, we ask whether frontotemporal cortex is functionally dissociated into distributed lexical and category-specific semantic networks. To this end, fMRI activation patterns elicited during the processing of words from different semantic categories were categorized using k-means cluster algorithms. Results showed a distributed pattern of inferiorfrontal, superiortemporal, and fusiform activation shared by different word categories. This shared activation contrasted with patterns of category-specific semantic activation in widely distributed neural systems. Clustering revealed congruent functional specificity of focal area activations in frontal and temporal cortex; thus suggesting a correspondence between functional partitionings of frontocentral mirror neuron systems and those of inferiortemporal lexical and semantic circuits. Action words related to the face, arms, and legs specifically activated the motor system in a somatotopic manner, whereas form-related words activated prefrontal areas. Similar functional specificity was evident in temporal cortex, where a different semantic topography emerged for form- and action-related words. Results were replicated in a separate data set, therefore recommending fMRI cluster analysis as a reliable method for scrutinizing the brain basis of lexical, semantic, and conceptual systems in humans. As focal modules do not explain the distributed character of functionally specific clusters and their distinct topographies are at variance with general distributed processing accounts, the functionally-homogenous distributed clusters specific to semantic types are best explained by specifically-distributed cortical circuits which, similar to Hebbian cell assemblies, represent functional units with specific roles in cognitive processing, especially in lexical and semantic access and memory. PMID- 19554562 TI - Contradiction in universal and particular reasoning. AB - A wide range of essential reasoning tasks rely on contradiction identification, a cornerstone of human rationality, communication and debate founded on the inversion of the logical operators "Every" and "Some." A high-density electroencephalographic (EEG) study was performed in 11 normal young adults. The cerebral network involved in the identification of contradiction included the orbito-frontal and anterior-cingulate cortices and the temporo-polar cortices. The event-related dynamic of this network showed an early negative deflection lasting 500 ms after sentence presentation. This was followed by a positive deflection lasting 1.5 s, which was different for the two logical operators. A lesser degree of network activation (either in neuron number or their level of phase locking or both) occurred while processing statements with "Some," suggesting that this was a relatively simpler scenario with one example to be figured out, instead of the many examples or the absence of a counterexample searched for while processing statements with "Every." A self-generated reward system seemed to resonate the recruited circuitry when the contradictory task is successfully completed. PMID- 19554561 TI - Mapping brain abnormalities in boys with autism. AB - Children with autism spectrum disorder (ASD) exhibit characteristic cognitive and behavioral differences, but no systematic pattern of neuroanatomical differences has been consistently found. Recent neurodevelopmental models posit an abnormal early surge in subcortical white matter growth in at least some autistic children, perhaps normalizing by adulthood, but other studies report subcortical white matter deficits. To investigate the profile of these alterations in 3D, we mapped brain volumetric differences using a relatively new method, tensor-based morphometry. 3D T1-weighted brain MRIs of 24 male children with ASD (age: 9.5 years +/- 3.2 SD) and 26 age-matched healthy controls (age: 10.3 +/- 2.4 SD) were fluidly registered to match a common anatomical template. Autistic children had significantly enlarged frontal lobes (by 3.6% on the left and 5.1% on the right), and all other lobes of the brain were enlarged significantly, or at trend level. By analyzing the applied deformations statistically point-by-point, we detected significant gray matter volume deficits in bilateral parietal, left temporal and left occipital lobes (P = 0.038, corrected), trend-level cerebral white matter volume excesses, and volume deficits in the cerebellar vermis, adjacent to volume excesses in other cerebellar regions. This profile of excesses and deficits in adjacent regions may (1) indicate impaired neuronal connectivity, resulting from aberrant myelination and/or an inflammatory process, and (2) help to understand inconsistent findings of regional brain tissue excesses and deficits in autism. PMID- 19554563 TI - Toward safe genetically modified organisms through the chemical diversification of nucleic acids. AB - It is argued that genetic proliferation should be rationally extended so as to enable the propagation in vivo of additional types of nucleic acids (XNA for 'xeno-nucleic acids'), whose chemical backbone motifs would differ from deoxyribose and ribose, and whose polymerization would not interfere with DNA and RNA biosynthesis. Because XNA building blocks do not occur in nature, they would have to be synthesized and supplied to cells which would be equipped with an appropriate enzymatic machinery for polymerizing them. The invasion of plants and animals with XNA replicons can be envisioned in the long run, but it is in microorganisms, and more specifically in bacteria, that the feasibility of such chemical systems and the establishment of genetic enclaves separated from DNA and RNA is more likely to take place. The introduction of expanded coding through additional or alternative pairing will be facilitated by the propagation of replicons based on alternative backbone motifs and leaving groups, as enabled by XNA polymerases purposefully evolved to this end. PMID- 19554564 TI - A surface-charge study on cellular-uptake behavior of F3-peptide-conjugated iron oxide nanoparticles. AB - Surface-charge measurements of mammalian cells in terms of Zeta potential are demonstrated as a useful biological characteristic in identifying cellular interactions with specific nanomaterials. A theoretical model of the changes in Zeta potential of cells after incubation with nanoparticles is established to predict the possible patterns of Zeta-potential change to reveal the binding and internalization effects. The experimental results show a distinct pattern of Zeta potential change that allows the discrimination of human normal breast epithelial cells (MCF-10A) from human cancer breast epithelial cells (MCF-7) when the cells are incubated with dextran coated iron oxide nanoparticles that contain tumor homing F3 peptides, where the tumor-homing F3 peptide specifically bound to nucleolin receptors that are overexpressed in cancer breast cells. PMID- 19554565 TI - High-order multiple emulsions formed in poly(dimethylsiloxane) microfluidics. PMID- 19554566 TI - Spatial deficits in a virtual water maze in amnesic participants with hippocampal damage. AB - The Morris water maze is a standard paradigm for the testing of hippocampal function in laboratory animals. Virtual versions of the Morris water maze are now available and can be used to assess spatial learning and memory ability in both healthy and brain injured participants. To evaluate the importance of the hippocampus in spatial learning and memory, we tested five amnesic participants with selective hippocampal damage using a virtual water maze called the Arena Maze. The amnesic participants with hippocampal damage were impaired on the invisible platform (place) task that required them to use distal cues, but were able to navigate almost as well as comparison participants when the invisible platform was marked by a single proximal cue. These results not only confirm that the hippocampus plays a necessary role in human navigation in large-scale environments but also provides a new link between the mnemonic and navigational roles of the hippocampus. PMID- 19554567 TI - Grp94, the endoplasmic reticulum Hsp90, has a similar solution conformation to cytosolic Hsp90 in the absence of nucleotide. AB - The molecular chaperone, Hsp90, is an essential eukaryotic protein that assists in the maturation and activation of client proteins. Hsp90 function depends upon the binding and hydrolysis of ATP, which causes large conformational rearrangements in the chaperone. Hsp90 is highly conserved from bacteria to eukaryotes, and similar nucleotide-dependent conformations have been demonstrated for the bacterial, yeast, and human proteins. There are, however, important species-specific differences in the ability of nucleotide to shift the conformation from one state to another. Although the role of nucleotide in conformation has been well studied for the cytosolic yeast and human proteins, the conformations found in the absence of nucleotide are less well understood. In contrast to cytosolic Hsp90, crystal structures of the endoplasmic reticulum homolog, Grp94, show the same conformation in the presence of both ADP and AMPPNP. This conformation differs from the yeast AMPPNP-bound crystal state, suggesting that Grp94 may have a different conformational cycle. In this study, we use small angle X-ray scattering and rigid body modeling to study the nucleotide free states of cytosolic yeast and human Hsp90s, as well as mouse Grp94. We show that all three proteins adopt an extended, chair-like conformation distinct from the extended conformation observed for the bacterial Hsp90. For Grp94, we also show that nucleotide causes a small shift toward the crystal state, although the extended state persists as the major population. These results provide the first evidence that Grp94 shares a conformational state with other Hsp90 homologs. PMID- 19554568 TI - Blind docking of pharmaceutically relevant compounds using RosettaLigand. AB - It is difficult to properly validate algorithms that dock a small molecule ligand into its protein receptor using data from the public domain: the predictions are not blind because the correct binding mode is already known, and public test cases may not be representative of compounds of interest such as drug leads. Here, we use private data from a real drug discovery program to carry out a blind evaluation of the RosettaLigand docking methodology and find that its performance is on average comparable with that of the best commercially available current small molecule docking programs. The strength of RosettaLigand is the use of the Rosetta sampling methodology to simultaneously optimize protein sidechain, protein backbone and ligand degrees of freedom; the extensive benchmark test described here identifies shortcomings in other aspects of the protocol and suggests clear routes to improving the method. PMID- 19554569 TI - Deep brain stimulation effects on gait variability in Parkinson's disease. AB - The effects of subthalamic nucleus (STN) deep brain stimulation (DBS) on fall risk in patients with Parkinson's disease (PD) currently remain unclear. Although several gait parameters, such as gait speed, have shown improvement with DBS, some studies have reported an increased fall risk following DBS. The purpose of this study was to examine the effect of bilateral DBS on gait variability, a marker of fall risk. The gait of 13 patients with idiopathic PD was analyzed to determine the influence of DBS, levodopa and both therapies together. Following treatment with both levodopa and STN DBS, subjects displayed improved gait speed, reduced gait variability (enhanced stability), and lower Unified Parkinson's Disease Rating Scale (UPDRS) scores. Although UPDRS scores improved with STN DBS alone, parallel improvements were not seen for gait variability. These findings suggest that different mechanisms may contribute to performance on UPDRS motor testing and gait stability in response to DBS. PMID- 19554570 TI - Initial testing (stage 1) of the kinesin spindle protein inhibitor ispinesib by the pediatric preclinical testing program. AB - BACKGROUND: Ispinesib is a highly specific inhibitor of kinesin spindle protein (KSP, HsEg5), a mitotic kinesin required for separation of the spindle poles. Here we report the activity of ispinesib against the in vitro and in vivo panels of the Pediatric Preclinical Testing Program (PPTP). PROCEDURES: Ispinesib was tested against the PPTP in vitro panel cell lines at concentrations from 0.1 nM to 1 microM and against the in vivo tumor panel xenografts by intraperitoneal administration (5 or 10 mg/kg) every 4 days for 3 doses repeated at day 21. RESULTS: Ispinesib was highly potent against the PPTP's in vitro cell lines with a median IC(50) of 4.1 nM. Ispinesib (10 mg/kg) induced unexplained toxicity in mice bearing osteosarcoma xenografts and exceeded the MTD in 12 of 40 non osteosarcoma xenografts. Ispinesib induced significant tumor growth delay in 88% (23/26) of evaluable xenografts. Using a time to event measure of efficacy, ispinesib had intermediate and high levels of activity against 4 (21%) and 5 (26%) of the 19 evaluable solid tumor xenografts, respectively. Ispinesib induced maintained complete responses (CR) in a rhabdoid tumor, a Wilms tumor and a Ewing sarcoma xenograft. Ispinesib (5 mg/kg) produced 2 complete and 2 partial responses among 6 evaluable xenografts in the ALL panel. The in vivo pattern of activity was distinctive from that previously reported for vincristine. CONCLUSIONS: Ispinesib demonstrated broad in vivo antitumor activity, including maintained complete responses for several xenografts, although with high toxicity rates at the doses studied. PMID- 19554571 TI - Initial testing (stage 1) of lapatinib by the pediatric preclinical testing program. AB - BACKGROUND: Lapatinib is a small molecule reversible tyrosine kinase inhibitor of EGFR and ErbB2 that shows in vitro and in vivo activity against a range of EGFR and ErbB2-dependent adult cancer cell lines and that has clinical efficacy against ErbB2-overexpressing breast cancer. METHODS: Lapatinib was tested against the cell lines of the PPTP in vitro panel at concentrations ranging from 1.0 nM to 10.0 microM. Lapatinib was tested against the xenografts of the PPTP in vivo panels using a twice-daily oral administration schedule for 6 weeks (5 days on, 2 days off) at a dose of 160 mg/kg (320 mg/kg/day). Lapatinib pharmacokinetic parameters were determined in scid(-/-) mice. RESULTS: The median IC(50) value for lapatinib against the entire PPTP cell line panel was 6.84 microM (range, 2.08 to >10.0 microM). Lapatinib was well tolerated in vivo, with toxicity in only 1.5% of the treated animals. Lapatinib induced significant differences in EFS distribution compared to controls in 1 of 41 xenografts tested. No objective responses were observed in any of the solid tumor panels or in the ALL panel. Lapatinib systemic exposure was consistent with previously observed values. CONCLUSIONS: Lapatinib has little activity against the xenografts of the PPTP's in vivo panel, and its in vitro activity occurs at concentrations above those associated with specific EGFR/ErbB2 inhibition. These results likely reflect lack of ErbB2 overexpression in the models studied and suggest that adult and pediatric cancers may fundamentally differ in the applicability of EGFR family members as therapeutic targets. PMID- 19554572 TI - Epigenetic inactivation of Homeobox A5 gene in nonsmall cell lung cancer and its relationship with clinicopathological features. AB - Promoter methylation is an important mechanism in gene silencing and is a key epigenetic event in cancer development. Homeobox A5 (HOXA5) is a master regulator of the morphogenesis and cell differentiation to be implicated as a tumor suppressor gene in breast cancer, but its role in lung cancer is still unknown. In this study, we have investigated the methylation status of the promoter region of the HOXA5 gene in nonsmall cell lung cancers (NSCLCs) using nested and standard methylation-specific PCR (MSP) and correlated the methylation status with clinicopathological features. With standard MSP analysis, HOXA5 methylation were found in 113 (81.3%) of 139 NSCLCs and 72 (51.8%) in their corresponding nonmalignant lung tissues. RT-PCR and immunohistochemical analysis showed that HOXA5 methylation correlates with gene expression. Moreover, in the patients with stage I disease, HOXA5 methylation was more frequent in smokers than in never smokes (P = 0.01). There was no influence of HOXA5 methylation on survival in all NSCLCs or at stages II-IV. However, in the patients with stage I disease, HOXA5 methylation was associated with a borderline significantly worse survival (P = 0.09). These findings suggest that downregulation of the HOXA5 gene by aberrant promoter methylation occurs in the vast majority of NSCLCs and that it may play a role in the pathogenesis of NSCLC. Additional studies with larger sample sizes are required to evaluate the prognostic value of HOXA5 methylation in patients with stage I NSCLC. PMID- 19554573 TI - Two genetic variants in prostate stem cell antigen and gastric cancer susceptibility in a Chinese population. AB - Genetic factors play important roles in pathogenesis of human cancer. A recent genome-wide association study (GWAS) linked two single nucleotide polymorphisms (SNPs) in prostate stem cell antigen (PSCA), rs2294008C>T and rs2976392G>A, to risk of diffuse-type of gastric cancer in Japanese and Korean populations. We hypothesized that these two SNPs are also associated with risk of gastric cancer in Chinese population. We examined genotypes and haplotypes of PSCA, rs2294008C/T and rs2976392G/A in 716 patients with cardia gastric carcinoma (CGC), 1020 patients with noncardia gastric carcinoma (NCGC), and 1020 controls. We found that individuals with at least one copy of the rs2294008T allele (CT or TT genotype) had an elevated risk for developing NCGC compared with those without this allele (OR = 1.35, 95% CI = 1.13-1.61). Individuals with at least one copy of the rs2976392A allele (GA or AA genotype) had nonsignificantly increased risk for NCGC compared with those without this allele (OR = 1.20, 95% CI = 1.01-1.43). Stratification analysis showed that the increased risk associated with the SNPs was restricted in female subjects. Moreover, the rs2294008T and rs2976392A allele carriers were predisposed to developing poorly differentiated and high stage NCGC at diagnosis. However, no such association was detected for CGC. In addition, we observed considerably lower allelic and genotype frequencies of these genetic variants in Chinese population compared with Japanese and Korean populations. These findings are in general consistent with previous GWAS and suggest that PSCA may play a role in the development of NCGC in Chinese population. PMID- 19554574 TI - Imaging single enzyme molecules under in situ conditions. PMID- 19554575 TI - Stereochemistry determination by powder X-ray diffraction analysis and NMR spectroscopy residual dipolar couplings. PMID- 19554576 TI - Highly substituted furo[3,4-d][1,2]oxazines: gold-catalyzed regiospecific and diastereoselective 1,3-dipolar cycloaddition of 2-(1-alkynyl)-2-alken-1-ones with nitrones. PMID- 19554577 TI - Nacnac... are you still there? The evolution of beta-diketiminate complexes of nickel. PMID- 19554578 TI - Methods for the incorporation of carbon-11 to generate radiopharmaceuticals for PET imaging. PMID- 19554579 TI - Induced-fit encapsulation by a 1,3,5-alternate calix[6]arene. PMID- 19554580 TI - Radicals and transition-metal catalysis: an alliance par excellence to increase reactivity and selectivity in organic chemistry. PMID- 19554581 TI - Ultrasensitive detection of bacteria using core-shell nanoparticles and an NMR filter system. PMID- 19554582 TI - Direct synthesis of iron, cobalt, nickel, and copper complexes of N-heterocyclic carbenes by using commercially available metal powders. PMID- 19554583 TI - Asymmetric total synthesis of soraphen A: a flexible alkyne strategy. PMID- 19554584 TI - Asymmetric self- and cross-aldol reactions of glycolaldehyde catalyzed by D fructose-6-phosphate aldolase. PMID- 19554585 TI - Chemoselective asymmetric N-allylic alkylation of indoles with Morita-Baylis Hillman carbonates. PMID- 19554586 TI - A short Bi=Bi bond supported by a metalloid group 13 ligand. PMID- 19554587 TI - On the role of metal contaminants in catalyses with FeCl3. PMID- 19554588 TI - Organocatalytic regio- and stereoselective inverse-electron-demand aza-Diels Alder reaction of alpha,beta-unsaturated aldehydes and N-tosyl-1-aza-1,3 butadienes. PMID- 19554589 TI - Copper-catalyzed cross-couplings with part-per-million catalyst loadings. PMID- 19554590 TI - Total synthesis of celogentin C. PMID- 19554591 TI - Development of a fluorescent peptide for the detection of vascular endothelial growth factor (VEGF). PMID- 19554592 TI - Potent triple helix stabilization by 5',3'-modified triplex-forming oligonucleotides. AB - Anthraquinone and pyrene analogues attached to the 3' and/or 5' termini of triplex-forming oligonucleotides (TFOs) by various linkers increased the stability of parallel triple helices. The modifications are simple to synthesize and can be introduced during standard solid-phase oligonucleotide synthesis. Potent triplex stability was achieved by using doubly modified TFOs, which in the most favourable cases gave an increase in melting temperature of 30 degrees C over the unmodified counterparts and maintained their selectivity for the correct target duplex. Such TFOs can produce triplexes with melting temperatures of 40 degrees C at pH 7 even though they do not contain any triplex-stabilizing base analogues. These studies have implications for the design of triplex-forming oligonucleotides for use in biology and nanotechnology. PMID- 19554594 TI - How can folded biopolymers and synthetic foldamers recognize each other? PMID- 19554593 TI - New, highly active nonbenzoquinone geldanamycin derivatives by using mutasynthesis. PMID- 19554596 TI - Dynamic transport in Li-conductive polymer electrolytes plasticized with poly(ethylene glycol)-borate/aluminate ester. AB - The addition of plasticizers into Li(+)-conductive solid polymer electrolytes (SPEs) is a commonly known technique to enhance the ionic conductivity. Among the used plasticizers, alkoxides of group-13 elements [such as poly(ethylene glycol) (PEG)-borate ester] are promising candidates due to the Lewis acidity of the elements of this group (i.e. B, Al, and so on), which interact with the anions and may increase the degree of dissociation of the salts and the transport number of the SPEs. By means of pulsed-gradient stimulated-echo NMR (PGStE-NMR) and AC impedance measurements, we investigate the effect of Lewis acidity originated from group-13 elements on the transport number and the dissociation rate of SPEs containing various plasticizers. Our results show that the degree of salt dissociation is significantly enhanced by the addition of plasticizers including group-13 elements, whereas only a small or negligible increase of the transport number is observed for these SPEs. We infer that the plasticizers exhibiting Lewis acidity associate with the anions, and that the associated pairs can migrate in the SPEs as fast as free anions, which results in a lower transport number than expected. PMID- 19554595 TI - DNA monofunctionalization of quantum dots. PMID- 19554597 TI - Reversible molecular logic: a photophysical example of a Feynman gate. PMID- 19554598 TI - Super-resolution imaging of DNA labelled with intercalating dyes. PMID- 19554600 TI - Synthesis, crystal structure and immunosuppressive activity of acylamide derivatives containing 1,4-benzodioxan. PMID- 19554599 TI - Structure-property relationships of a class of carbamate-based fatty acid amide hydrolase (FAAH) inhibitors: chemical and biological stability. AB - Cyclohexylcarbamic acid aryl esters are a class of fatty acid amide hydrolase (FAAH) inhibitors, which includes the reference compound URB597. The reactivity of their carbamate fragment is involved in pharmacological activity and may affect their pharmacokinetic and toxicological properties. We conducted in vitro stability experiments in chemical and biological environments to investigate the structure-stability relationships in this class of compounds. The results show that electrophilicity of the carbamate influences chemical stability, as suggested by the relation between the rate constant of alkaline hydrolysis (log k(pH9)) and the energy of the lowest unoccupied molecular orbital (LUMO). Introduction of small electron-donor substituents at conjugated positions of the O-aryl moiety increased the overall hydrolytic stability of the carbamate group without affecting FAAH inhibitory potency, whereas peripheral non-conjugated hydrophilic groups, which favor FAAH recognition, helped decrease oxidative metabolism in the liver. PMID- 19554601 TI - Truxene-based columnar liquid crystals: self-assembled structures and electro active properties. AB - Columnar liquid-crystalline (LC) truxene derivatives containing branched flexible alkyl chains have been designed and synthesized. The dicyanomethylene and dithiafulvene substituents have been introduced into the pi-conjugated truxene framework to tune their electronic and redox properties as well as the molecular assembled structures. The pi-conjugated cores of dicyanomethylene- and dithiafulvene-appended truxenes adopt bowl-shaped conformations, giving rise to a large intrinsic dipole moment perpendicular to the aromatic framework. These molecules form stable columnar LC structures through intermolecular dipole-dipole interactions. The redox properties of LC truxene derivatives have been examined by cyclic voltammetry. The dicyanomethylene-appended truxene shows the reversible four-step electrochemical reductions, whereas the dithiafulvene-appended truxene undergoes three-step oxidations. PMID- 19554602 TI - Ge pairs and Sb ribbons in rare-earth germanium antimonides RE12Ge(7-x)Sb21 (RE = La-Pr). AB - The ternary rare-earth germanium antimonides RE(12)Ge(7-x)Sb(21) (RE = La-Pr; x = 0.4-0.5) are synthesized by direct reactions of the elements. Single-crystal X ray diffraction studies indicate that they adopt a new structure type (space group Immm, Z = 2, a = 4.3165(4)-4.2578(2) A, b = 15.2050(12)-14.9777(7) A, c = 34.443(3)-33.9376(16) A in the progression from RE = La to Pr), integrating complex features found in RE(6)Ge(5-x)Sb(11+x) and RE(12)Ga(4)Sb(23). A three dimensional polyanionic framework, consisting of Ge pairs and Sb ribbons, outlines large channels occupied by columns of face-sharing RE(6) trigonal prisms. These trigonal prisms are centered by additional Ge and Sb atoms to form GeSb(3) trigonal-planar units. A bonding analysis attempted through a Zintl-Klemm approach suggests that full electron transfer from the RE atoms to the anionic substructure cannot be assumed. This is confirmed by band-structure calculations, which also reveal the importance of Ge-Sb and Sb-Sb bonding. Magnetic measurements on Ce(12)Ge(6.5)Sb(21) indicate antiferromagnetic coupling but no long-range ordering down to 2 K. PMID- 19554603 TI - A zwitterionic silylene as reactive intermediate and its unusual dimerization to an N-heterobicyclic disilane. AB - A way to synthesize the transient zwitterionic silylene L'Si: 8 {L'=CH[(C=CH(2))CMe(N(tBu))(2)]} and achieve its facile dimerization to the remarkable N-heterobicyclic disilane 8(2) is described. At first, employing the beta-diketiminate ligand L [L=CH(CMeN(tBu))(2)], both starting materials LH (2) and its N-lithium salt LLi (3) can react with SiBr(4) to yield the silylene precursor L'SiBr(2) (4) by silicon-induced C-H activation at an exocyclic methyl group on the backbone of the ligand. Compound 4 reacts with SiBr(4) above room temperature to afford the unexpected terminal CH(SiBr(3))-substituted dibromosilane 6 along with the unique tricyclic trisilane 7. Reduction of 4 with KC(8) at 0 degrees C furnishes the novel N-heterobicyclic disilane 8(2), which is a formal dimer of the desired zwitterionic silylene L'Si: (8). It has been reasoned that compound 8(2) may results from [4+1] cycloaddition of two molecules of 8 to give the transient dimer 8(2)', which subsequently undergoes hydrogen transfer from a terminal methyl group on the backbone of the C(3)N(2)Si ligand to the low-coordinate Si atom. The latter dimerization can be rationalized by the intrinsic zwitterionic character of 8 and insufficient steric protection through the tBu groups at the nitrogen atoms. The novel compounds 3, 4, 6, 7, and 8(2) have been characterized by (1)H, (13)C, and (29)Si NMR spectroscopy, mass spectrometry, and elemental analysis. Additionally, the structures of 3, 6, 7, and 8(2) were also established by single-crystal X-ray diffraction analyses. PMID- 19554604 TI - NH/pi attraction: a role in asymmetric hydrogenation of aromatic ketones with binap/1,2-diamine-ruthenium(II) complexes. PMID- 19554605 TI - Carbon-supported, selenium-modified ruthenium-molybdenum catalysts for oxygen reduction in acidic media. AB - The stability and oxygen reduction activity of two carbon-supported catalyst materials are reported. The catalysts, Se/Ru and Se/(Ru-Mo), were prepared by using a chemical reduction method. The catalyst nanoparticles were evenly dispersed onto globular amorphous carbon supports, and their average size was ca. 2.4 nm. Thermal treatment at 500 degrees C for 2 h in an inert argon atmosphere resulted in coarsening of the nanoparticles, and also in some decrease of their activity. A gradual reduction of activity was also observed for Se/Ru during potential-cycle experiments. However, the incorporation of small amounts of Mo into the Se/Ru catalysts considerably improved the stability of the catalyst against dissolution. The Mo-containing samples showed excellent oxygen reduction activities even after cycling the potential 1000 times between 0.7 and 0.9 V. Furthermore, they showed excellent fuel-cell behavior. The performance of the Se/Ru catalysts is greatly improved by the addition of small amounts of elemental Mo. Possible mechanisms responsible for the improvement of the activity are discussed. PMID- 19554606 TI - Acid-catalyzed conversion of sugars and furfurals in an ionic-liquid phase. AB - The reactivity of monosaccharides, furfural, and 5-hydroxymethyl-2-furfural (HMF) in the presence of a Bronsted acid (added as H(2)SO(4)) in the ionic liquid 1 butyl-3-methylimidazolium chloride (BMImCl) is investigated at 120 degrees C. Fructose is converted much faster than mannose, glucose, and xylose and yields HMF with high selectivity, even in the absence of acid. Conversion of mannose, glucose, and xylose involves more complex reaction networks. Only small amounts of furfural and HMF are converted in the absence of other reactants but both compounds are consumed when monosaccharides and their degradation products are present. Acid-catalyzed degradation reactions also lead to the formation of solid residues (humins). PMID- 19554607 TI - Sustainable management of the global carbon cycle through geostorage of wood. AB - Combustion of fossil energy sources has caused the carbon inventory of the atmosphere to increase by more than 200 Gt. It will be almost impossible to prevent it from growing by at least another 400 Gt in the present century. Theoretically, there exists only one single possibility to effect a decline of the resultant increase in atmospheric CO(2) concentration: the excess carbon has to be removed from the carbon cycle by transferring it into an environment in which it is safe from oxidation, just as is the case for the deposits of fossil fuels. Only natural photosynthesis offers the possibility of efficiently fixing carbon dioxide from the air and removing it from the carbon cycle through geostorage of the resulting biomass. The present paper shows, in the context of an initial feasibility study, that the use of forests and the geostorage of wood in an environment corresponding to lignite deposits represents the ecologically most sensible and economical variant of removal of carbon from the carbon cycle and, thereby, reclamation of the atmosphere. PMID- 19554608 TI - Nitrite reduction: a ubiquitous function from a pre-aerobic past. AB - In eukaryotes, small amounts of nitrite confer cytoprotection against ischemia/reperfusion-related tissue damage in vivo, possibly via reduction to nitric oxide (NO) and inhibition of mitochondrial function. Several hemeproteins are involved in this protective mechanism, starting with deoxyhemoglobin, which is capable of reducing nitrite. In facultative aerobic bacteria, such as Pseudomonas aeruginosa, nitrite is reduced to NO by specialized heme-containing enzymes called cd(1) nitrite reductases. The details of their catalytic mechanism are summarized below, together with a hypothesis on the biological role of the unusual d(1)-heme, which, in the reduced state, shows unique properties (very high affinity for nitrite and exceptionally fast dissociation of NO). Our results support the idea that the nitrite-based reactions of contemporary eukaryotes are a vestige of earlier bacterial biochemical pathways. The evidence that nitrite reductase activities of enzymes with different cellular roles and biochemical features still exist today highlights the importance of nitrite in cellular homeostasis. PMID- 19554609 TI - The physiological role of hormones in saliva. AB - The assessment of hormones in saliva has gained wide acceptance in clinical endocrinology. To date, there is no hypothesis as to why some hormones can be found in saliva, while others cannot, and whether there is a physiological consequence of this fact. A number of carefully performed studies give examples of important physiological hormonal activity in saliva. Steroids, such as androgens, act as pheromones in olfactory communication of various mammalian species, such as facilitating mating behavior in swine or serving as odor cues for rodent nestlings. Salivary peptide hormones, such as epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha), and amines such as melatonin, are involved in the regulation of inflammatory processes and in the promotion of cell proliferation, and contribute to a rapid wound healing in the oropharyngeal epithelia. Current data provide evidence of the involvement of salivary cytokines, such as interleukin-8 and leptin, in tumorgenesis in the oral cavity and the salivary glands. The tumor tissues express and release significantly more of these cytokines than healthy glands. Consequently, the assessment of salivary hormone profiles may provide promising targets for diagnostic tumor markers. PMID- 19554610 TI - Regulation, necessity, and the misinterpretation of knockouts. AB - Much contemporary biology consists of identifying the molecular components that associate to perform biological functions, then discovering how these functions are controlled. The concept of control is key to biological understanding, at least of the physiological kind; identifying regulators of processes underpins ideas of causality and allows complicated, multicomponent systems to be summarized in relatively simple diagrams and models. Unfortunately, as this article demonstrates by drawing on published articles, there is a growing tendency for authors to claim that a molecule is a 'regulator' of something on evidence that cannot support the conclusion. In particular, gene knockout experiments, which can demonstrate only that a molecule is necessary for a process, are all too frequently being misinterpreted as revealing regulation. This logical error threatens to blur the important distinction between regulation and mere necessity and therefore to weaken one of our strongest tools for comprehending how organisms work. PMID- 19554611 TI - Polycomb group proteins: remembering how to catch chromatin during replication. AB - Polycomb group (PcG) proteins maintain the expression state of PcG-responsive genes during development of multicellular organisms. Recent observations suggest that "the H3K27me3 modification" acts to maintain Polycomb repressive complex (PRC) 2, the enzyme that creates this modification, on replicating chromatin. This could in turn promote propagation of H3K27me3 on newly replicated daughter chromatin, and promote recruitment of PRC1. Other work suggests that PRC1-class complexes can be maintained on replicating chromatin, at least in vitro, independently of H3K27me3. Thus, histone modifications and PcG proteins themselves may both be maintained through replication. PMID- 19554613 TI - Bipartite vectors for co-expression of a growth factor cDNA and short hairpin RNA against an apoptotic gene. AB - BACKGROUND: Although human islet transplantation is a promising approach for treating type I diabetes, its success is limited as a result of the poor survival rate of transplanted islets. Expression of a growth factor gene to promote revascularization and silencing of pro-apoptotic genes before transplantation may improve the outcome of islet transplantation. METHODS: In the present study, we constructed bipartite plasmid vectors to co-express a vascular endothelial growth factor (VEGF) cDNA and short hairpin (sh)RNA targeting inducible NO synthase (iNOS) gene. First, we screened shRNA sequences against human iNOS by transfecting plasmids encoding shRNA targeting different start sites of human iNOS. Then, the effect of different promoters [such as H1, U6 and cytomegalovirus (CMV)] and micro RNA backbones on gene silencing was determined. RESULTS: No statistical difference in iNOS gene silencing was observed for the shRNA with H1, U6 and CMV promoters. In addition, a conventional shRNA showed better silencing of the iNOS gene compared to shRNA containing mir375 and mir30 backbones. A bipartite plasmid was also constructed with mir30-shRNA and a VEGF cDNA controlled by a single CMV promoter. This plasmid showed a better silencing effect compared to plasmid without VEGF cDNA. CONCLUSIONS: In the present study, we have successfully constructed bipartite vectors co-expressing a VEGF cDNA and a shRNA against the iNOS gene. These vectors could be attractive candidates for improving the survival of transplanted islets. PMID- 19554612 TI - APOE mRNA and protein expression in postmortem brain are modulated by an extended haplotype structure. AB - Currently the epsilon4 allele of the apolipoprotein E gene (APOE) is the strongest genetic risk factor for late onset Alzheimer's disease (AD). However, inheritance of the APOE epsilon4 allele is not necessary or sufficient for the development of AD. Genetic evidence suggests that multiple loci in a 70 kb region surrounding APOE are associated with AD risk. Even though these loci could represent surrogate markers in linkage disequilibrium with APOE epsilon4 allele, they could also contribute biological effects independent of the APOE epsilon4 allele. Our previous study identified multiple SNPs upstream from APOE that are associated with cerebrospinal fluid apoE levels, suggesting that a haplotype structure proximal to APOE can influence apoE expression. In this study, we examined apoE expression in human post-mortem brain (PMB), and constructed chromosome-phase-separated haplotypes of the APOE proximal region to evaluate their effect on PMB apoE expression. ApoE protein expression was found to differ among AD brain regions and to differ between AD and control hippocampus. In addition, an extended APOE proximal haplotype structure, spanning from the TOMM40 gene to the APOE promoter, may modulate apoE expression in a brain region specific manner and may influence AD disease status. In conclusion, this haplotype-phenotype analysis of apoE expression in PMB suggests that either; (1) the cis-regulation of APOE expression levels extends far upstream of the APOE promoter or (2) an APOE epsilon4 allele independent mechanism involving the TOMM40 gene plays a role in the risk of AD. PMID- 19554615 TI - Reproducibility and validity of digital inclinometry for measuring cervical range of motion in normal subjects. AB - BACKGROUND AND PURPOSE: Measurements of cervical range of motion (CROM) have been extensively reported in the past decade employing simple (goniometers) as well as sophisticated (electro-, magneto- and ultrasonography-based) systems. The recent introduction of the simple, user-friendly and relatively cheap digital inclinometer (DI) has opened a potentially new venue for measurement of this segment's motion. The purpose of the present study was to assess intra-tester reproducibility of DI-based findings as well as its validity in comparison to the ultrasonography-based Zebris CMS 70P (Zebris Medizintechnik Gmbh, Isny, Germany) for measuring CROM in normal subjects. METHODS: Active CROM of healthy women (n = 15) and men (n = 15) aged 24.2(2.4) years was measured on two sessions, Test 1 and Test 2, spread over 7.2(+/-0.7) days apart. On Test 1, the six primary movements of the neck (flexion, F; extension, E; right and left lateral flexion, RLF and LLF; and right and left rotations, RR and LR) were measured using the DI and the Zebris. On Test 2, the same measurements were performed using the DI only. All measurements were conducted by the same tester, with the subject in the seated position. The only exception was DI measurement of cervical rotation that was performed in the supine position due to the DI gravity-dependence, rendering DI measurements in the transverse plane irrelevant. RESULTS: No significant differences were revealed between the two instruments with respect to the sagittal and frontal planes, whereas the DI-based CROM in rotation was significantly greater then its Zebris-based counterpart. The inter-device interclass correlation coefficients (ICCs) for the frontal were 0.72 (RLF) and 0.62 (LLF), and 0.77 (F) and 0.83 (E). Poor correlations were indicated for the rotations. The intra-tester reproducibility derived from the test-retest DI measurement indicated good to excellent reproducibility in all planes with ICCs ranging from 0.82 (LLF) to 0.94 (E). The Standard Error of Measurement ranged from 1.6 degrees (RR) to 2.6 degrees (F). CONCLUSION: DI-based CROM measurements are reproducible and valid for recording sagittal and frontal plane motions in healthy subjects. The higher range in rotations, relative to the Zebris-based findings, is most probably attributable to the test position. Being relatively cheap, portable and convenient for tester and subject alike, the DI seems to be an effective instrument for assessing CROM. PMID- 19554614 TI - Association analyses between brain-expressed fatty-acid binding protein (FABP) genes and schizophrenia and bipolar disorder. AB - Deficits in prepulse inhibition (PPI) are a biological marker for psychiatric illnesses such as schizophrenia and bipolar disorder. To unravel PPI-controlling mechanisms, we previously performed quantitative trait loci (QTL) analysis in mice, and identified Fabp7, that encodes a brain-type fatty acid binding protein (Fabp), as a causative gene. In that study, human FABP7 showed genetic association with schizophrenia. FABPs constitute a gene family, of which members FABP5 and FABP3 are also expressed in the brain. These FABP proteins are molecular chaperons for polyunsaturated fatty acids (PUFAs) such as arachidonic and docosahexaenoic acids. Additionally, the involvement of PUFAs has been documented in the pathophysiology of schizophrenia and mood disorders. Therefore in this study, we examined the genetic roles of FABP5 and 3 in schizophrenia (N = 1,900 in combination with controls) and FABP7, 5, and 3 in bipolar disorder (N = 1,762 in the case-control set). Three single nucleotide polymorphisms (SNPs) from FABP7 showed nominal association with bipolar disorder, and haplotypes of the same gene showed empirical associations with bipolar disorder even after correction of multiple testing. We could not perform association studies on FABP5, due to the lack of informative SNPs. FABP3 displayed no association with either disease. Each FABP is relatively small and it is assumed that there are multiple regulatory elements that control gene expression. Therefore, future identification of unknown regulatory elements will be necessary to make a more detailed analysis of their genetic contribution to mental illnesses. PMID- 19554616 TI - Relationship of cranial robusticity to cranial form, geography and climate in Homo sapiens. AB - Variation in cranial robusticity among modern human populations is widely acknowledged but not well-understood. While the use of "robust" cranial traits in hominin systematics and phylogeny suggests that these characters are strongly heritable, this hypothesis has not been tested. Alternatively, cranial robusticity may be a response to differences in diet/mastication or it may be an adaptation to cold, harsh environments. This study quantifies the distribution of cranial robusticity in 14 geographically widespread human populations, and correlates this variation with climatic variables, neutral genetic distances, cranial size, and cranial shape. With the exception of the occipital torus region, all traits were positively correlated with each other, suggesting that they should not be treated as individual characters. While males are more robust than females within each of the populations, among the independent variables (cranial shape, size, climate, and neutral genetic distances), only shape is significantly correlated with inter-population differences in robusticity. Two block partial least-squares analysis was used to explore the relationship between cranial shape (captured by three-dimensional landmark data) and robusticity across individuals. Weak support was found for the hypothesis that robusticity was related to mastication as the shape associated with greater robusticity was similar to that described for groups that ate harder-to-process diets. Specifically, crania with more prognathic faces, expanded glabellar and occipital regions, and (slightly) longer skulls were more robust than those with rounder vaults and more orthognathic faces. However, groups with more mechanically demanding diets (hunter-gatherers) were not always more robust than groups practicing some form of agriculture. PMID- 19554617 TI - Joint orientation and function in great ape and human proximal pedal phalanges. AB - Previous studies have referred to the degree of dorsal canting of the base of the proximal phalanx as an indicator of human-like metatarsophalangeal joint function and thus a diagnostic trait of habitual bipedality in the fossil record. Here, we used a simple method to investigate differences in forefoot function on a finer scale. Building on Duncan et al.'s (Am J Phys Anthropol 93 [1994] 67-81) research, we tested whether dorsal canting reflects differences between sexes in locomotor behavior, whether habitual shoe wear influences dorsal canting in humans, and whether proximal joint morphology differs between rays in Pan and humans. Our results corroborate previous research in showing that humans have proximal phalanges with joint orientations that are significantly more dorsal than, but overlap with, those of great apes. We also found that male gorillas have significantly more dorsally canted second proximal phalanges than their female counterparts, while the opposite pattern between the sexes was found in Pan troglodytes. Inter-ray comparisons indicate that Pan have more dorsally canted first proximal phalanges than second proximal phalanges, while the opposite pattern was found in humans. Minimally shod humans have slightly but significantly more dorsally canted second proximal phalanges than those of habitually shod humans, indicating that phalanges of unshod humans provide the most appropriate comparative samples for analyses of early hominins. Overall, our analysis suggests that though the measurement of dorsal canting is limited in its sensitivity to certain intraspecific differences in function, phalangeal joint orientation reflects interspecific differences in joint function, with the caveat that different patterns of forefoot function during gait can involve similar articular sets of metatarsophalangeal joints. PMID- 19554618 TI - Hepatology: a home for hepatocellular cancer publications. PMID- 19554620 TI - Is this dystonia? AB - Torsion dystonia is characterized by sustained muscle contractions causing twisting and repetitive movements and abnormal postures. The diagnosis can be made difficult, delayed, and often misled by several factors: variability of dystonia presentation, uncertain recognition of the specific physical signs, lack of diagnostic tests, wide etiological spectrum, and coexistence of other movement disorders. Diagnostic tools are of limited assistance for the diagnosis of dystonia, which remains based on clinical diagnostic skills. We propose here, a new diagnostic algorithm to systematize the clinical diagnostic workout. A correct recognition of the physical signs that constitute the hallmark of most dystonia syndromes provides the grounds to perform a structured diagnostic sequence and share a consistent methodology. This clinical algorithm may be enhanced by adding diagnostic tools for dystonia, once their diagnostic value is assessed. PMID- 19554619 TI - Health disparities in liver disease: Time to take notice and take action. PMID- 19554623 TI - Neuroprotection in a 6-hydroxydopamine-lesioned Parkinson model using lactoferrin modified nanoparticles. AB - BACKGROUND: Nonviral gene therapy of chronic degenerative diseases such as Parkinson's disease (PD) is a great challenge as a result of the low tranfection efficiency of nonviral gene vectors. We previously constructed a lactoferrin (Lf) modified vector, which was demonstrated to be potential for brain gene delivery both in vitro and in vivo. In the present study, this type of vector was applied to load human glial cell line-derived neurotrophic factor gene (hGDNF). METHODS: A rat PD model was constructed by the unilateral lesion of striatum using 6 hydroxydopamine (6-OHDA). Lf-modified nanoparticles (NPs) were prepared and characterized. Neuroprotective effects of Lf-modified NPs were examined in the 6 OHDA-lesioned PD model via a regimen of multiple dosing intravenous administrations. RESULTS: The size of Lf-modified NPs was 196 +/- 10.1 nm, whereas the zeta potential value was 29.35 +/- 3.27 mV. Lf-modified NPs could protect themselves from heparin displacement and DNase digestion. The results of the neuroprotective evaluation show that increasing the number of injections of Lf-modified NPs loading hGDNF improved locomotor activity, reduced dopaminergic neuronal loss and enhanced monoamine neurotransmitter levels in PD rats. Five injections of Lf-modified NPs loading hGDNF exhibited much more powerful neuroprotection than a single injection, indicating the effectiveness and feasibility of multiple dosing administrations. The results of toxicity tests demonstrated that the dosage of NPs used in the present study was safe enough for brain gene delivery. CONCLUSIONS: The findings obtained in the present study suggest that Lf-modified NPs could be developed for potential nonviral gene therapy of chronic brain disorders. PMID- 19554624 TI - Combining angiogenic gene and stem cell therapies for myocardial infarction. AB - BACKGROUND: Transplantation of stem cells from various sources into infarcted hearts has the potential to promote myocardial regeneration. However, the regenerative capacity is limited partly as a result of the low survival rate of the transplanted cells in the ischemic myocardium. In the present study, we tested the hypothesis that combining cell and angiogenic gene therapies would provide additive therapeutic effects via co-injection of bone marrow-derived mesenchymal stem cells (MSCs) with an adeno-associated viral vector (AAV), MLCVEGF, which expresses vascular endothelial growth factor (VEGF) in a cardiac specific and hypoxia-inducible manner. METHODS: MSCs isolated from transgenic mice expressing green fluorescent protein and MLCVEGF packaged in AAV serotype 1 capsid were injected into mouse hearts at the border of ischemic area, immediately after occlusion of the left anterior descending coronary, individually or together. Engrafted cells were detected and quantified by real time polymerase chain reaction and immunostaining. Angiogenesis and infarct size were analyzed on histological and immunohistochemical stained sections. Cardiac function was analyzed by echocardiography. RESULTS: We found that co-injection of AAV1-MLCVEGF with MSCs reduced cell loss. Although injection of MSCs and AAV1 MLCVEGF individually improved cardiac function and reduced infarct size, co injection of MSC and AAV1-MLCVEGF resulted in the best improvement in cardiac function as well as the smallest infarct among all groups. Moreover, injection of AAV1-MLCVEGF induced neovasculatures. Nonetheless, injection of MSCs attracted endogenous stem cell homing and increased scar thickness. CONCLUSIONS: Co injection of MLCVEGF and MSCs in ischemic hearts can result in better cardiac function and MSC survival, compared to their individual injections, as a result of the additive effects of each therapy. PMID- 19554625 TI - Antifibrotic effect through the regulation of transcription factor using ring type-Sp1 decoy oligodeoxynucleotide in carbon tetrachloride-induced liver fibrosis. AB - BACKGROUND: Liver fibrosis is characterized by the excessive accumulation of extracellular matrix (ECM). Recent advances in the knowledge about the cellular, molecular and genetic aspects of fibrosis have opened a new era of research on liver cirrhosis. A transcription factor, Sp1, originally described as a ubiquitous transcription factor, is involved in the basal expression of ECM genes and may be important in the fibrotic processes. METHODS: The chronic hepatic damage received intraperitoneal injection of carbon tetrachloride (2 mg/kg) dissolved in corn oil (1 : 3 ratio) three times a weekly for 8 weeks. The delivery of decoy oligodeoxynucleotide (ODN) was performed by injection of 10 microg of scrambled decoy ODN or 10 microg of ring type (R)-Sp1 decoy ODN through the mouse tail vein. All animals of each group were sacrificed, DNA binding activity, expression of cytokines and histological analysis were measured. RESULTS: We have generated a R-Sp1 decoy ODN that effectively blocks Sp1 binding to the promoter region for transcription regulation of transforming growth factor (TGF)-beta1. The expression of fibrotic cytokines and inflammatory cytokines was decreased by using the R-Sp1 decoy ODN in liver cirrhosis. CONCLUSIONS: The present study demonstrates that the R-Sp1 decoy ODN inhibits TGF-beta1 expression in liver cirrhosis. These results indicate that targeting Sp1 can efficiently block ECM expression, and suggest that such an approach may represent an interesting therapeutic alternative towards the treatment of cirrhosis. PMID- 19554626 TI - Determination and application of empirically derived detergent phase boundaries to effectively crystallize membrane proteins. AB - Elucidating the structures of membrane proteins is essential to our understanding of disease states and a critical component in the rational design of drugs. Structural characterization of a membrane protein begins with its detergent solubilization from the lipid bilayer and its purification within a functionally stable protein-detergent complex (PDC). Crystallization of the PDC typically occurs by changing the solution environment to decrease solubility and promote interactions between exposed hydrophilic surface residues. As membrane proteins have been observed to form crystals close to the phase separation boundaries of the detergent used to form the PDC, knowledge of these boundaries under different chemical conditions provides a foundation to rationally design crystallization screens. We have carried out dye-based detergent phase partitioning studies using different combinations of 10 polyethylene glycols (PEG), 11 salts, and 11 detergents to generate a significant amount of chemically diverse phase boundary data. The resulting curves were used to guide the formulation of a 1536-cocktail crystallization screen for membrane proteins. We are making both the experimentally derived phase boundary data and the 1536 membrane screen available through the high-throughput crystallization facility located at the Hauptman Woodward Institute. The phase boundary data have been packaged into an interactive Excel spreadsheet that allows investigators to formulate grid screens near a given phase boundary for a particular detergent. The 1536 membrane screen has been applied to 12 membrane proteins of unknown structures supplied by the structural genomics and structural biology communities, with crystallization leads for 10/12 samples and verification of one crystal using X-ray diffraction. PMID- 19554627 TI - Structural characterization of alpha-synuclein in an aggregation prone state. AB - The relation of alpha-synuclein (alphaS) aggregation to Parkinson's disease has long been recognized, but the pathogenic species and its molecular properties have yet to be identified. To obtain insight into the properties of alphaS in an aggregation-prone state, we studied the structural properties of alphaS at acidic pH using NMR spectroscopy and computation. NMR demonstrated that alphaS remains natively unfolded at lower pH, but secondary structure propensities were changed in proximity to acidic residues. The ensemble of conformations of alphaS at acidic pH is characterized by a rigidification and compaction of the Asp and Glu rich C-terminal region, an increased probability for proximity between the NAC region and the C-terminal region and a lower probability for interactions between the N- and C-terminal regions. PMID- 19554628 TI - Detection of early unfolding events in a dimeric protein by amide proton exchange and native electrospray mass spectrometry. AB - Oligomeric proteins generally undergo unfolding through a dissociation/denaturation mechanism wherein the subunits first dissociate and then unfold. This mechanism can be detected by the fact that the proteins exhibit a concentration dependence of the denaturation curve. However, the concentration dependence does not answer the question of whether there are thermally induced conformational changes that facilitate subunit dissociation. To fully probe these mechanisms it is desirable to have an analytical approach that is capable of measuring both subunit dissociation and protein denaturation in a highly sensitive manner. In this article, we demonstrate that the combined use of native mass spectrometry to detect subunit mixing, and amide hydrogen/deuterium exchange to detect transient unfolding events can provide a very unique insight into the pre-melting transitions in a protein oligomer. Both methods keep an isotopic record of each transformation event, without the dependence on equilibrium of the unfolding reaction. Here, we use a combined form of H/D exchange/mass spectrometry and isotopic labeling/native electrospray mass spectrometry to study the pre-unfolding events of Bacillus subtilis NAD(+) synthetase, a symmetrical dimer protein, which plays a vital role in the lifecycle of the bacteria. In the experimental outcome provided, we were able to clearly illustrate that at elevated temperatures, the NAD synthetase dimer undergoes reversible dissociation without monomer unfolding, while at temperatures where monomer unfolding is observed to take place, the rate of dimer dissociation still yet exceeds the rate of unfolding. Information provided by combining these two mass spectrometric methods was found to be very robust, and allowed us to establish an NAD synthetase unfolding model, where primary dissociation occurs prior to the complete unfolding of the NAD(+) synthetase. PMID- 19554629 TI - Solution structure of the phytotoxic protein PcF: the first characterized member of the Phytophthora PcF toxin family. AB - The PcF protein from Phytophthora cactorum is the first member of the "PcF toxin family" from the plant pathogens Phytophthora spp. It is able to induce withering in tomato and strawberry leaves. The lack of sequence similarity with other proteins hampers the identification of the molecular mechanisms responsible for its toxicity. Here, we show that the six cysteines form a disulphide pattern that is exclusive for PcF and essential for the protein withering activity. The NMR solution structure identifies a novel fold among protein effectors: a helix-loop helix motif. The presence of a negatively charged surface suggests that it might act as a site of electrostatic interaction. Interestingly, a good fold match with Ole e 6, a plant protein with allergenic activity, highlighted the spatial superimposition of a stretch of identical residues. This finding suggests a possible biological activity based on molecular mimicry. PMID- 19554630 TI - Castration induces autoantibody and T cell responses that correlate with inferior outcomes in an androgen-dependent murine tumor model. AB - We recently reported that hormone therapy induces antigen-specific autoantibody responses in prostate cancer patients. However, the contribution of autoantibody responses to clinical outcomes is unknown. We used an animal model to test the hypothesis that hormone therapy-induced immune responses may be associated with delayed tumor recurrence. Male DD/S mice bearing established tumors from the androgen-dependent Shionogi carcinoma line were castrated to induce tumor regression. Tumor-specific autoantibody responses were measured by immunoblot, and the underlying antigen was identified by serological screening of a cDNA expression library. T cell responses were assessed by immunohistochemistry and IFN-gamma ELISPOT. Following castration, 97% of mice underwent complete tumor regression. Of these, 72% experienced tumor recurrence 18-79 days postcastration, whereas the remaining 28% remained tumor-free for the duration of the experiment. In 55% of mice, castration induced autoantibody responses to an antigen identified as poly(A) binding protein nuclear 1 (PABPN1). Castration also induced PABPN1-specific T cell responses, which were highly correlated to autoantibody responses, and this was accompanied by dense infiltration of tumors by CD3+ T cells 1-2 weeks after castration. Unexpectedly, mice that developed autoantibody and T cell responses to PABPN1 showed a higher rate and shorter latency of tumor recurrence. In mice with recurrent tumors, T cell responses to PABPN1 were still detectable; however, T cell infiltrates were restricted to the peripheral stroma of tumors. In conclusion, castration-induced immune responses are associated with inferior outcomes in the Shionogi carcinoma model, raising concerns about the influence of treatment-induced immune responses on clinical outcomes in humans. PMID- 19554631 TI - Safety and efficacy of the endoscopic modified Lothrop procedure: a systematic review and meta-analysis. AB - OBJECTIVES/HYPOTHESIS: The endoscopic modified Lothrop procedure (EMLP; also known as Draf III or frontal drillout) has recently gained popularity as a minimally invasive alternative to frontal sinus obliteration. This systematic analysis was designed to assess the safety and efficacy of the EMLP. STUDY DESIGN: Literature review and meta-analysis. METHODS: We performed a search of all English studies published from 1990 to 2008 that reported results from a minimum of five patients undergoing the EMLP. Of the 33 papers reviewed, 18 studies (evidence level II-2 or II-3) containing data from 612 patients met inclusion criteria. RESULTS: The most common indications for EMLP were chronic frontal sinusitis (75.2%) and mucocele (21.3%). Patients had an average age of 47.9 years (range, 14-89 years) and were followed for 28.5 months postoperatively. Only 20.3% of procedures were performed without image-guidance. Stents were rarely used (6%). The rate of major and minor complications was <1% and 4%, respectively. No deaths were reported. A majority of patients were discharged within 24 hours. Postoperative endoscopic findings, qualitatively reported in 394 patients, demonstrated frontal sinus patency or partial stenosis in 95.9% at last follow-up. Where specifically assessed (n = 430 patients), improvement in symptoms was achieved in 82.2% of cases, with 16% reporting no significant change, and 1.2% reporting worsening of symptoms. The overall failure rate (requiring further surgery) of EMLP was 13.9% (85/612). Of the failures, 80% underwent revision EMLP, whereas 20% elected osteoplastic frontal sinus obliteration. CONCLUSIONS: When performed by an experienced surgeon, EMLP is a safe and efficacious procedure that is well tolerated. PMID- 19554632 TI - Evaluation of hand circulation before radial forearm free flap surgery. AB - The radial forearm free flap (RFFF) is a versatile flap offering many reconstructive options for head and neck defects. Its low bulk, ease of dissection, and excellent vascularity are among its advantages. Its low flap loss and complication rates offer the best choice for the reconstruction of oral, oropharyngeal, and hypopharyngeal lining. Evaluation of collateral circulation to the hand is important to avoid distal ischemia and donor site morbidity. Pulse oximetry can be used to augment Allen's test to assess collateral supply to the hand. This cost-neutral technique can be used in routine preoperative screening prior to RFFF surgery. PMID- 19554634 TI - An underreported complication of laryngeal microdebrider: vocal fold web and granuloma: a case report. AB - OBJECTIVES/HYPOTHESIS: The microdebrider has become a valuable instrument for otolaryngologists. It is now used in the larynx for treatment of recurrent respiratory papillomatosis, laryngeal stenosis, and debridement of large cancers for airway control. There are few reported complications reported with the use of the microdebrider in the larynx. STUDY DESIGN: A case report. METHODS: A patient with a vocal fold polyp underwent removal of the polyp with a microdebrider at an outside institution. He presented to our clinic 2 months after the excision with a severely strained near aphonic voice. On rigid stroboscopic examination we saw a large anterior commisure laryngeal web with a granuloma. This was repaired by granuloma excision, web lysis, buccal graft, and laryngeal stent placement. CONCLUSIONS: The microdebrider is an extremely valuable tool for the otolaryngologist. Violation of the epithelium and the lamina propria with muscle exposure can result in serious damage to the vocal folds. When using powered instrumentation the surgeon should use the upmost caution in the larynx to avoid causing debilitating injury and scar with subsequent dysphonia. PMID- 19554633 TI - A new treatment option for laryngeal sensory neuropathy. AB - OBJECTIVES/HYPOTHESIS: Laryngeal sensory neuropathy (LSN) may produce a variety of symptoms, including chronic cough, globus sensation, odynophonia, and/or odynophagia. Etiologies are often iatrogenic, viral, or idiopathic, although the diagnosis is generally one of exclusion. The aim of this study is to introduce pregabalin (Lyrica, Pfizer Inc., New York, NY) as a potential new therapy for LSN. STUDY DESIGN: Retrospective clinical investigation. METHODS: : Charts were reviewed from 12 consecutive patients who were prescribed pregabalin for symptoms of LSN. Outcomes were reviewed by analyzing pre and post-treatment questionnaires asking patients to rate symptoms on a scale from 0 to 5. Adverse effects and evidence of drug tolerance were also recorded. RESULTS: Two patients did not tolerate pregabalin due to somnolence. Of those that tolerated the medication, mean pretreatment chief complaint symptom severity rating was 3.9, whereas mean post-treatment symptom rating was 1.2 after 1 month of pregabalin therapy. None of the patients developed drug tolerance effects over time. CONCLUSIONS: Pregabalin therapy appears to be an effective treatment option for laryngeal sensory neuropathy. Future prospective studies are needed to compare outcomes between pregabalin and other medications as treatments for LSN. PMID- 19554635 TI - Low COX2 in tumor and upregulation in stroma mark laryngeal squamous cell carcinoma progression. AB - OBJECTIVES/HYPOTHESIS: Invasive squamous cell carcinomas (SCC) of the larynx, like most solid tumors, are surrounded by a reactive stroma, in which cancer associated fibroblasts (CAFs) are the predominant cell type. This mesenchymal reaction may affect cancer progression multiply. The proinflammatory enzyme cyclooxygenase-2 (COX-2) has been correlated with head and neck cancer. This study aims to explore the impact of epithelial and stromal COX-2 expression on SCC behavior. STUDY DESIGN: Retrospective case review study performed in a tertiary health center institution. METHODS: Double immunohistochemistry of COX-2 and the CAF marker alpha-smooth muscle actin (alpha-SMA) was utilized in 97 laryngeal cancer patients. Follow-up data were collected in 52 cases. RESULTS: Low COX-2 immunostaining in cancer cells was associated with advanced grade (P = .044) and shorter recurrence-free period (P = .035). CAF expression was positively correlated with the grade of the infiltrating tumor (P = .030). CONCLUSIONS: In laryngeal SCCs, COX-2 may exert its deleterious effect by alterations in the tumor microenvironment. CAF-derived, COX-2-mediated paracrine influences on malignant cells possibly facilitate cancer progression. Overlooking the stromal remodeling could account for unsuccessful treatments of epithelial neoplasms. PMID- 19554636 TI - Differentiating between adductor and abductor spasmodic dysphonia using airflow interruption. AB - OBJECTIVES/HYPOTHESIS: To measure the laryngeal resistance (R(L)), subglottal pressure (P(s)), and mean flow rate (MFR) of adductor (ADSD) and abductor (ABSD) spasmodic dysphonia patients using the airflow interrupter. METHODS: The R(L) of six ABSD and seven ADSD patients was measured using the airflow interrupter, a noninvasive device designed to measure MFR and P(s) via mechanical balloon valve interruption. Subjects performed 10 trials at each of two intensity levels, with each trial consisting of a sustained /a/ during which phonation was interrupted for 500 ms. Laryngeal resistance was calculated as subglottal pressure divided by airflow. RESULTS: Mean R(L) for the ADSD and ABSD subtypes at 65 dB were 24.78 cmH(2)O/L/s and 14.51 cmH(2)O/L/s, respectively (P = .04). Mean R(L) at 70 dB were 40.02 cmH(2)O/L/s and 15.84 cmH(2)O/L/s (P = .014). P(s) for the ADSD and ABSD subtypes at 65 dB were 10.23 cmH(2)O and 8.32 cmH(2)O, respectively (P = .582). At the 70 dB level, P(s) were 12.39 cmH(2)O and 11.78 cmH(2)O (P = .886). MFR for the ADSD and ABSD subtypes at 65 dB were 435 mL/s and 746 mL/s (P = .205). Mean MFR at 70 dB were 518 mL/s and 848 mL/s (P = .198). CONCLUSIONS: Noninvasive measurements of R(L) may be useful for differentiating between ADSD and ABSD. This simple objective test, which produces a quantitative output, could be used to evaluate laryngeal function in patients with spasmodic dysphonia. PMID- 19554637 TI - Lemierre's syndrome: A systematic review. AB - OBJECTIVES/HYPOTHESIS: Lemierre's syndrome is characterized by a history of recent oropharyngeal infection, clinical or radiological evidence of internal jugular vein thrombosis, and isolation of anaerobic pathogens, mainly Fusobacterium necrophorum. It was once called the forgotten disease because of its rarity, but it may not be that uncommon after all. This review aims to provide physicians with an update on the etiology, management, and prognosis of Lemierre's syndrome. METHODS: Systematic review using the terms: Lemierre's syndrome, postanginal septicemia, fusobacterium, internal jugular vein thrombosis. INCLUSION CRITERIA: English literature; reviews, case reports, and case series. EXCLUSION CRITERIA: variants or atypical Lemierre's syndrome cases, negative fusobacteria cultures, and papers without radiological evidence of thrombophlebitis. RESULTS: Eighty-four studies fulfilled our inclusion criteria. The male to female ratio was 1:1, 2, and the ages ranged from 2 months to 78 years (median, 22 years). Main sources of infection were tonsil, pharynx, and chest. Most common first clinical presentation was a sore throat, followed by a neck mass and neck pain. The most common offending micro-organism was F. necrophorum. Treatment modalities used were antimicrobial, anticoagulant, and surgical treatment. Morbidity was significant with prolonged hospitalization in the majority of patients. The overall mortality rate was 5%. CONCLUSIONS: Lemierre's syndrome may not be as rare as previously thought. This apparent increase in the incidence may be due to antibiotic resistance or changes in antibiotic prescription patterns. Successful management rests on the awareness of the condition, a high index of suspicion, and a multidisciplinary team approach. PMID- 19554638 TI - 5-Ethynyl-2'-deoxyuridine labeling detects proliferating cells in the regenerating avian cochlea. AB - OBJECTIVES/HYPOTHESIS: The avian cochlea regenerates hair cells following aminoglycoside treatment through supporting cell proliferation. Immunocytochemical labeling of 5-bromo-2'-deoxyuridine (BrdU), a thymidine analog, is a popular nonradioactive marker for identifying cells in the DNA synthesis (S phase) of the cell cycle. However, it requires harsh treatments to denature double-stranded DNA for the antibody to bind BrdU. We explored a new method using 5-ethynyl-2'-deoxyuridine (EdU) as a thymidine analog and a nonantibody azide/alkyne reaction between EdU and the fluorescent probe. We propose that EdU is as effective as BrdU, but without the requirement for harsh denaturation or the use of antibodies for detection. STUDY DESIGN: Two-week-old chicks received a single gentamicin injection followed by a single EdU injection 72 hours later. Cochleae were extracted 4-8 hours later, fixed, and processed for fluorescent detection of EdU. METHODS: Cochleae were processed for detection of incorporated EdU using the Click-iT Imaging Kit (Invitrogen/Molecular Probes, Carlsbad, CA) and colabeled with Sox2, myosin VI, or myosin VIIa antibodies. Whole-mount cochlear preparations were examined with confocal microscopy. RESULTS: Supporting cells incorporated EdU into their newly synthesized DNA during the 4-8 hours following the EdU injection and were readily detected with little background signal. The intensity and quantity of cells labeled were similar to or better than that seen for BrdU. CONCLUSIONS: The EdU method is as effective as BrdU, without requiring harsh denaturation or secondary antibodies to identify proliferating cells. Thus, the nonantibody EdU system allows more flexibility by enabling colabeling with multiple antibodies to other cellular proteins involved in regeneration. PMID- 19554639 TI - Endoscopic repair of laryngeal cleft type I and type II: when and why? AB - OBJECTIVES/HYPOTHESIS: To evaluate the clinical features of children with type I and type II laryngeal cleft and the role of conservative monitoring versus endoscopic repair in their management. METHODS: Clinical presentation and evaluation; findings at the time of laryngoscopy, bronchoscopy, and esophagoscopy; and efficacy and outcome of conservative monitoring and endoscopic CO(2) laser repair. RESULTS: Eighty-one patients were evaluated for aspiration. Seventy-four patients were diagnosed as having a clinically significant laryngeal cleft. Thirty-two patients (14 males, 18 females) were monitored conservatively. Forty-nine patients (26 males, 23 females) required surgical intervention due to failed medical and feeding therapy of aspiration related to their laryngeal clefts (28 type I, 21 type II). Endoscopic CO(2) laser repair was used in all these patients. CONCLUSIONS: Medical and feeding therapy should be the first modality of treatment in patients with laryngeal cleft type I and type II. Factors supporting surgical repair include: 1) clinically apparent aspiration with feeding, 2) severity of pulmonary status, 3) findings on modified barium swallow and chest x-ray, 4) absence of significant comorbid conditions predisposing to aspiration, 5) findings on upper aerodigestive endoscopy, and 6) poor response to medical management and feeding therapy. PMID- 19554640 TI - Endoscopic transnasal approach to the clivus: a radiographic anatomical study. AB - OBJECTIVES/HYPOTHESIS: Operative intervention of anterior skull base lesions is challenging. Various endoscopic surgical approaches have been described. The goal of the present study is to perform a radiographic analysis of the endoscopic transnasal approach to the clivus. STUDY DESIGN: Anatomic study utilizing computed tomography (CT). METHODS: High-resolution surgical-guidance CT images of the sinuses from 97 patients at a tertiary care medical center between 2002 and 2007 were evaluated. Axial and sagittal images were used to evaluate surgical access to the clivus. Multiple anatomical measurements were obtained and analyzed with imaging and statistical software. RESULTS: Of the 97 imaging studies, there were 39 males and 58 females. The width of exposure of the clivus without removal of the septum was 2.7 cm (1.9-3.4 cm) and with removal of the bony septum was 3.6 cm (2.6-4.8 cm) (P < .001). No patients had complete exposure of the width of the clivus without the septum removed compared to 56 (58%) patients with the septum removed. Endoscopic exposure of the inferior and superior limits of the clivus was not limited in any images studied. CONCLUSIONS: The endoscopic transnasal approach to the clivus is a viable option in the treatment of anterior skull base lesions with the preservation of functional anatomy in select patients. A large portion of the population has limited lateral exposure secondary to the eustachian tube and the medial pterygoid plate with an endoscopic transnasal approach. Vertically, this approach allows complete access to the clivus in all patients studied. PMID- 19554641 TI - Outpatient repair of superior semicircular canal dehiscence via the transmastoid approach. AB - OBJECTIVES/HYPOTHESIS: Superior semicircular canal dehiscence (SSCD) syndrome has been a topic of much interest since its first description a decade ago. The symptoms of vertigo, autophony, and Tullio phenomenon have been well described as has the utility of surgical repair. The standard approach described for surgical repair of this problem has been to perform a middle fossa craniotomy followed by plugging of the superior semicircular canal. Recently, a transmastoid approach has been described as another surgical option in cases of SSCD, an alternative that could avoid the known risks of a middle fossa craniotomy. Herein we present further data for validation of the transmastoid SSCD repair technique. Additional factors leading to the successful treatment of these patients in the outpatient setting, an approach not previously described, are detailed. STUDY DESIGN: Retrospective study of three separate operative procedures in two patients (one patient with bilateral SSCD) in an academic medical center is presented along with postoperative course and hearing status. METHODS: Three ears with radiographic evidence of SSCD confirmed with vestibular evoked myogenic potentials after symptomatic presentation were studied. Each ear had preoperative and postoperative audiometry and outpatient surgery. SSCD was repaired in each patient using a transmastoid approach with specific anesthetic and surgical precautions taken to minimize nausea and vertigo. RESULTS: Each patient was discharged from the outpatient unit with two cases returning home the day of surgery and one case after 23-hour observation. All had resolution of their SSCD symptoms and postoperative hearing testing revealed no evidence of sensorineural hearing loss and resolution of related conductive components. CONCLUSIONS: The advantages noted by others with regards to the transmastoid repair of SSCD related symptomatology appear genuine and reproducible. In addition, this approach appears to offer potential freedom from a prolonged hospital admission with the potential for outpatient surgery in some circumstances without compromised clinical outcomes. PMID- 19554642 TI - Trabecular microarchitecture of hominoid thoracic vertebrae. AB - Spontaneous vertebral fractures are a common occurrence in modern humans, yet these fractures are not documented in other hominoids. Differences in vertebral bone strength between humans and apes associated with trabecular bone microarchitecture may contribute to differences in fracture incidence. We used microcomputed tomography to examine trabecular bone microarchitecture in the T8 vertebra of extant young adult hominoids. Scaled volumes of interest from the anterior vertebral body were analyzed at a resolution of 46 microm, and bone volume fraction, trabecular thickness, trabecular number, trabecular separation, structure model index, and degree of anisotropy were compared among species. As body mass increased, so did trabecular thickness, but bone volume fraction, structure model index, and degree of anisotropy were independent of body mass. Bone volume fraction was not significantly different between the species. Degree of anisotropy was not significantly different among the species, suggesting similarity of loading patterns in the T8 vertebra due to similar anatomical and postural relationships within each species' spine. Degree of anisotropy was negatively correlated with bone volume fraction (r(2) = 0.85, P < 0.05) in humans, whereas the apes demonstrated no such relationship. This suggested that less dense human trabecular bone was more preferentially aligned to habitual loading. Furthermore, we theorize that trabeculae in ape thoracic vertebrae would not be expected to become preferentially aligned if bone volume fraction was decreased. The differing relationship between bone volume fraction and degree of anisotropy in humans and apes may cause less dense human bone to be more fragile than less dense ape bone. PMID- 19554643 TI - The velocity-related firing property of hippocampal place cells is dependent on self-movement. AB - Hippocampal place cells have the interesting property of increasing their firing rate when a freely moving animal increases its running speed through the cell's place field. A previous study from this laboratory showed that this movement related firing property is disrupted by lesions of the perirhinal cortex (PrhC). It is possible, therefore, that PrhC lesions disrupt speed-modulated sensory information such as optic flow or motor efferent or proprioceptive input that might be available to the hippocampus from the PrhC. To test this hypothesis, rats with single unit recording electrodes implanted in the CA1 region of the hippocampus received different levels of optic flow stimulation in both a freely moving and a passive movement condition. The effects of PrhC lesions were also tested. Although increasing the amount of optic flow information available decreased place field size, it had no discernable effect on the movement-firing rate relationship in the place cells of control animals run in the free-movement condition. In lesioned animals the relationship was disrupted, replicating our previous results. In the passive movement condition many place cells stopped firing. In those cells that did fire, however, the movement-firing rate relationship was no longer evident. These data indicate that the movement-firing rate relationship is not driven by vestibular or optic flow cues, but rather depends on either motor efferent or proprioceptive input, or that it results from some other form of input that may be modulated by self-motion, such as from the vibrissae. PMID- 19554645 TI - Adult learning and remodeling of hippocampal mossy fibers: unheralded participant in circuitry for long-lasting spatial memory. AB - Two articles in this issue concerning the overexpression of GAP-43 on mossy fiber growth are related to the plasticity of these axons in relation to learning and memory. PMID- 19554644 TI - Alcohol inhibition of neurogenesis: a mechanism of hippocampal neurodegeneration in an adolescent alcohol abuse model. AB - Adolescents diagnosed with an alcohol use disorder show neurodegeneration in the hippocampus, a region important for learning, memory, and mood regulation. This study examines a potential mechanism by which excessive alcohol intake, characteristic of an alcohol use disorder, produces neurodegeneration. As hippocampal neural stem cells underlie ongoing neurogenesis, a phenomenon that contributes to hippocampal structure and function, we investigated aspects of cell death and cell birth in an adolescent rat model of an alcohol use disorder. Immunohistochemistry of various markers along with Bromo-deoxy-Uridine (BrdU) injections were used to examine different aspects of neurogenesis. After 4 days of binge alcohol exposure, neurogenesis was decreased by 33 and 28% at 0 and 2 days after the last dose according to doublecortin expression. To determine whether this decrease in neurogenesis was due to effects on neural stem cell proliferation, quantification of BrdU-labeled cells revealed a 21% decrease in the dentate gyrus of alcohol-exposed brains. Cell survival and phenotype of BrdU labeled cells were assessed 28 days after alcohol exposure and revealed a significant, 50% decrease in the number of surviving cells in the alcohol-exposed group. Reduced survival was supported by significant increases in the number of pyknotic-, FluoroJade B positive-, and TUNEL-positive cells. However, so few cells were TUNEL-positive that cell death is likely necrotic in this model. Although alcohol decreased the number of newborn cells, it did not affect the percentage of cells that matured into neurons (differentiation). Thus, our data support that in a model of an adolescent alcohol use disorder, neurogenesis is impaired by two mechanisms: alcohol-inhibition of neural stem cell proliferation and alcohol effects on new cell survival. Remarkably, alcohol inhibition of neurogenesis may outweigh the few dying cells per section, which implies that alcohol inhibition of neurogenesis contributes to hippocampal neurodegeneration in alcohol use disorders. PMID- 19554646 TI - Thermoresponsive, hydrolytically degradable polymer micelles intended for radionuclide delivery. AB - Novel polymer micelles, prepared by self-assembling thermoresponsive poly(N isopropylacrylamide)-graft-poly[N-(2-hydroxypropyl)methacrylamide] copolymers with hydrolytically degradable N-glycosylamine groups between the polymer blocks are proposed for delivery of diagnostic and therapeutic radionuclides into solid tumors. The micelles are formed by fast heating of an aqueous solution of the copolymer to 37 degrees C. They have a hydrodynamic diameter of 128 nm (measured using dynamic light scattering) and slowly degrade during incubation in aqueous buffer at pH = 7.4. Labeling with both (131)I and (90)Y proceeds with high yields (>85%). The unlabeled polymers are not cytotoxic for any of the tested murine and human cell lines. PMID- 19554647 TI - Acute pancreatitis at the beginning of the 21st century: the state of the art. AB - Acute pancreatitis is an acute inflammatory disease of the pancreas which can lead to a systemic inflammatory response syndrome with significant morbidity and mortality in 20% of patients. Gallstones and alcohol consumption are the most frequent causes of pancreatitis in adults. The treatment of mild acute pancreatitis is conservative and supportive; however severe episodes characterized by necrosis of the pancreatic tissue may require surgical intervention. Advanced understanding of the pathology, and increased interest in assessment of disease severity are the cornerstones of future management strategies of this complex and heterogeneous disease in the 21st century. PMID- 19554649 TI - Eosinophilic colitis. AB - Eosinophilic colitis (EC) is a rare form of primary eosinophilic gastrointestinal disease with a bimodal peak of prevalence in neonates and young adults. EC remains a little understood condition in contrast to the increasingly recognized eosinophilic esophagitis. Clinical presentation of EC is highly variable according to mucosal, transmural, or serosal predominance of inflammation. EC has a broad differential diagnosis because colon tissue eosinophilia often occurs in parasitic infection, drug-induced allergic reactions, inflammatory bowel disease, and various connective tissue disorders, which require thorough searching for secondary causes that may be specifically treated with antibiotics or dietary and drug elimination. Like eosinophilic gastrointestinal disease involving other segments of the gastrointestinal tract, EC responds very well to steroids that may be spared by using antihistamines, leukotriene inhibitors and biologics. PMID- 19554648 TI - Systemic abnormalities in liver disease. AB - Systemic abnormalities often occur in patients with liver disease. In particular, cardiopulmonary or renal diseases accompanied by advanced liver disease can be serious and may determine the quality of life and prognosis of patients. Therefore, both hepatologists and non-hepatologists should pay attention to such abnormalities in the management of patients with liver diseases. PMID- 19554650 TI - Psychosocial stress and liver disease status. AB - "Psychosocial stress" is an increasingly common concept in the challenging and highly-demanding modern society of today. Organic response to stress implicates two major components of the stress system, namely the hypothalamic-pituitary adrenal axis and the sympathetic nervous system. Stress is anamnestically reported by patients during the course of disease, usually accompanied by a decline in their overall health status. As the mechanisms involving glucocorticoids and catecholamines have been deciphered, and their actions on immune cell function deeper understood, it has become clear that stress has an impact on hepatic inflammatory response. An increasing number of articles have approached the link between psychosocial stress and the negative evolution of hepatic diseases. This article reviews a number of studies on both human populations and animal models performed in recent years, all linking stress, mainly of psychosocial nature, and the evolution of three important liver-related pathological entities: viral hepatitis, cirrhosis and hepatocellular carcinoma. PMID- 19554651 TI - Pluronic L-81 ameliorates diabetic symptoms in db/db mice through transcriptional regulation of microsomal triglyceride transfer protein. AB - AIM: To test whether oral L-81 treatment could improve the condition of mice with diabetes and to investigate how L-81 regulates microsomal triglyceride transfer protein (MTP) activity in the liver. METHODS: Genetically diabetic (db/db) mice were fed on chow supplemented with or without L-81 for 4 wk. The body weight, plasma glucose level, plasma lipid profile, and adipocyte volume of the db/db mice were assessed after treatment. Toxicity of L-81 was also evaluated. To understand the molecular mechanism, HepG2 cells were treated with L-81 and the effects on apolipoprotein B (apoB) secretion and mRNA level of the MTP gene were assessed. RESULTS: Treatment of db/db mice with L-81 significantly reduced and nearly normalized their body weight, hyperphagia and polydipsia. L-81 also markedly decreased the fasting plasma glucose level, improved glucose tolerance, and attenuated the elevated levels of plasma cholesterol and triglyceride. At the effective dosage, little toxicity was observed. Treatment of HepG2 cells with L 81 not only inhibited apoB secretion, but also significantly decreased the mRNA level of the MTP gene. Similar to the action of insulin, L-81 exerted its effect on the MTP promoter. CONCLUSION: L-81 represents a promising candidate in the development of a selective insulin-mimetic molecule and an anti-diabetic agent. PMID- 19554652 TI - Nanosized As2O3/Fe2O3 complexes combined with magnetic fluid hyperthermia selectively target liver cancer cells. AB - AIM: To study the methods of preparing the magnetic nano-microspheres of Fe(2)O(3) and As(2)O(3)/Fe(2)O(3) complexes and their therapeutic effects with magnetic fluid hyperthermia (MFH). METHODS: Nanospheres were prepared by chemical co-precipitation and their shape and diameter were observed. Hemolysis, micronucleus, cell viability, and LD(50) along with other in vivo tests were performed to evaluate the Fe(2)O(3) microsphere biocompatibility. The inhibition ratio of tumors after Fe(2)O(3) and As(2)O(3)/Fe(2)O(3) injections combined with induced hyperthermia in xenograft human hepatocarcinoma was calculated. RESULTS: Fe(2)O(3) and As(2)O(3)/Fe(2)O(3) particles were round with an average diameter of 20 nm and 100 nm as observed under transmission electron microscope. Upon exposure to an alternating magnetic field (AMF), the temperature of the suspension of magnetic particles increased to 41-51 degrees C, depending on different particle concentrations, and remained stable thereafter. Nanosized Fe(2)O(3) microspheres are a new kind of biomaterial without cytotoxic effects. The LD(50) of both Fe(2)O(3) and As(2)O(3)/Fe(2)O(3) in mice was higher than 5 g/kg. One to four weeks after Fe(2)O(3) and As(2)O(3)/Fe(2)O(3) complex injections into healthy pig livers, no significant differences were found in serum AST, ALT, BUN and Cr levels among the pigs of all groups (P > 0.05), and no obvious pathological alterations were observed. After exposure to alternating magnetic fields, the inhibition ratio of the tumors was significantly different from controls in the Fe(2)O(3) and As(2)O(3)/Fe(2)O(3) groups (68.74% and 82.79%, respectively; P < 0.01). Tumors of mice in treatment groups showed obvious necrosis, while normal tissues adjoining the tumor and internal organs did not. CONCLUSION: Fe(2)O(3) and As(2)O(3)/Fe(2)O(3) complexes exerted radiofrequency induced hyperthermia and drug toxicity on tumors without any liver or kidney damage. Therefore, nanospheres are ideal carriers for tumor-targeted therapy. PMID- 19554653 TI - Clinicopathological analysis of paraganglioma with literature review. AB - AIM: To investigate the 152 cases of paragangliomas resected over the past 32 years in West China Hospital clinicopathologically. METHODS: All cases of paragangliomas diagnosed at the Department of Gastrointestinal Surgery and Department of Pathology, West China Hospital, China were reviewed. The pathological documents were supplied by the Department of Pathology, West China Hospital, and other necessary data were extracted from the hospital records. The statistical analyses were performed by survival analysis (Kaplan-Meier method), descriptive statistical analyses and chi(2) analysis. RESULTS: The neuroendocrine marker vimentin was found to be selectively expressed in the benign tumors, and there were significant differences in the expression of those markers in both benign and malignant tumors. The survival analysis revealed that survival correlated significantly with the malignancy, metastasis and nodal status. CONCLUSION: Vimentin may be useful in the differential diagnosis between malignant and benign tumors. The difference in the expression of this marker in the tumors could be a clue to the future clinical diagnosis. The malignancy, metastasis and the nodal status may predict the prognosis of this disease. PMID- 19554654 TI - Influence of heme oxygenase-1 expression on immune liver fibrosis induced by cobalt protoporphyrin in rats. AB - AIM: To investigate the effect of heme oxygenase-1 (HO-1) expression on immune liver fibrosis induced by cobalt protoporphyrin (CoPP) in rats. METHODS: An immune liver fibrosis model of rat was established by administering human serum albumin (HSA). The rats were divided into CoPP, liver fibrosis and normal control groups. Rats in the CoPP group received intraperitoneal CoPP concurrently with HSA. Expression of HO-1 protein was observed by Western blotting and immunohistochemistry. Hematoxylin and eosin (HE) staining was performed to assess fibrosis proliferation and distribution, proliferation extent of fibroblasts, and alterations in hepatocytes and inflammatory cells. Type I and III collagens were detected with Van Gieson's (VG) staining and Foot's reticular fiber staining, respectively. In addition, spindle-shaped cells existing at perisinusoidal locations beyond portal and septa areas were investigated with HE staining. RESULTS: Western blotting and immunohistochemistry showed that the expression of HO-1 protein was higher in the CoPP group than in the liver fibrosis group (P < 0.05). Compared with the liver fibrosis group, the serological index of hepatic fibrosis in the CoPP group decreased significantly (P < 0.05). HE, VG and Foot's staining revealed that administration of CoPP reduced the extent of hepatic fibrosis. The levels of serological indicators and the number of spindle-shaped cells at perisinuous locations beyond the portal and septa areas were reduced in the CoPP group. Only a few inflammatory cells were seen around the portal areas and central veins in the CoPP group. CONCLUSION: Increased endogenous HO-1 may suppress liver fibrosis by protecting liver cells, inhibiting inflammatory cell infiltration and hepatic stellate cell transformation. PMID- 19554656 TI - Lower baseline ALT cut-off values and HBV DNA levels better differentiate HBeAg- chronic hepatitis B patients from inactive chronic carriers. AB - AIM: To determine whether new cut-off values for alanine aminotransferase (ALT) and baseline hepatitis B virus (HBV) DNA levels better differentiate HBeAg(-) chronic hepatitis B (CHB) patients from inactive chronic carriers. METHODS: Ninety-one patients [32 HBeAg(+) CHB, 19 inactive carriers and 40 HBeAg(-) CHB] were followed up for 2 years and were tested for HBV DNA levels by a PCR-based assay. ALT was tested twice during the last 6 mo using new cut-off values: ULN (upper limit of normal) 30 IU/L for males, 19 IU/L for females. Diagnostic accuracy, sensitivity, specificity, positive and negative predictive values were calculated by discriminant analysis. RESULTS: When using the revised ALT cut-off values, the lowest optimal HBV DNA level that differentiated HBeAg(-) CHB patients from inactive carriers was 50 000 copies/mL. The diagnostic accuracy of HBV DNA to determine inactive carriers with a cut-off of 50 000 copies/mL was similar to the previously recommended cut-off of 100 000 copies/mL (91%). HBV DNA levels were lower than the cut-off value in 95% of inactive carriers and in 28% of HBeAg(-) CHB patients. With ALT < 30 IU/L in men and < 19 IU/L in women and HBV DNA levels < 100 000 copies/mL, the risk of CHB is 5%. On the other hand, if ALT values were > 30 IU in men and > 19 IU in women and baseline HBV DNA levels were > 100 000 copies/mL, the risk is 86%. CONCLUSION: New cut-off values for ALT together with HBV DNA levels proposed by AASLD (American Association for the Study of Liver Diseases) and NIH (National Institute of Health) consensus seem appropriate to characterize inactive carriers. PMID- 19554655 TI - Survival predictors in patients treated with a molecular adsorbent recirculating system. AB - AIM: To identify prognostic factors for survival in patients with liver failure treated with a molecular adsorbent recirculating system (MARS). METHODS: MARS is a liver-assisting device that has been used in the treatment of liver failure to enable native liver recovery, and as a bridge to liver transplantation (LTX). We analyzed the 1-year outcomes of 188 patients treated with MARS, from 2001 to 2007, in an intensive care unit specializing in liver disease. Demographic, clinical and laboratory parameters were recorded before and after each treatment. One-year survival and the number of LTXs were recorded. Logistic regression analysis was performed to determine factors predicting survival. RESULTS: The study included 113 patients with acute liver failure (ALF), 62 with acute-on chronic liver failure (AOCLF), 11 with graft failure (GF), and six with miscellaneous liver failure. LTX was performed for 29% of patients with ALF, 18% with AOCLF and 55% with GF. The overall 1-year survival rate was 74% for ALF, 27% for AOCLF, and 73% for GF. The poorest survival rate, 6%, was noted in non transplanted patients with alcohol-related AOCLF and cirrhosis, whereas, patients with enlarged and steatotic liver had 55% survival. The etiology of liver failure was the most important predictor of survival (P < 0.0001). Other prognostic factors were encephalopathy (P = 0.001) in paracetamol-related ALF, coagulation factors (P = 0.049) and encephalopathy (P = 0.064) in non-paracetamol-related toxic ALF, and alanine aminotransferase (P = 0.013) and factor V levels (P = 0.022) in ALF of unknown etiology. CONCLUSION: The etiology of liver disease was the most important prognostic factor. MARS treatment appears to be ineffective in AOCLF with end-stage cirrhosis without an LTX option. PMID- 19554657 TI - Improving quality of colonoscopy by adding simethicone to sodium phosphate bowel preparation. AB - AIM: To evaluate the effectiveness of simethicone in enhancing visibility and efficacy during colonoscopy. METHODS: A prospective, double-blind, randomized, placebo-controlled study was conducted. One hundred and twenty-four patients were allocated to receive 2 doses of sodium phosphate plus 240 mg of tablet simethicone or placebo as bowel preparation. Visibility was blindly assessed for the amount of air bubbles and adequacy of colon preparation. Total colonoscopic time, side effects of the medication, endoscopist and patient satisfaction were also compared. RESULTS: Sodium phosphate plus simethicone, compared to sodium phosphate plus placebo, improved visibility by diminishing air bubbles (100.00% vs 42.37%, P < 0.0001) but simethicone failed to demonstrate improvement in adequacy of colon preparation (90.16% vs 81.36%, P = 0.17). Endoscopist and patient satisfaction were increased significantly in the simethicone group. However, there was no difference in the total duration of colonoscopy and side effects of the medication. CONCLUSION: The addition of simethicone is of benefit for colonoscopic bowel preparation by diminishing air bubbles, which results in enhanced visibility. Endoscopist and patient satisfaction is also increased. PMID- 19554658 TI - Acute extensive portal and mesenteric venous thrombosis after splenectomy: treated by interventional thrombolysis with transjugular approach. AB - AIM: To present a series of cases with symptomatic acute extensive portal vein (PV) and superior mesenteric vein (SMV) thrombosis after splenectomy treated by transjugular intrahepatic approach catheter-directed thrombolysis. METHODS: A total of 6 patients with acute extensive PV-SMV thrombosis after splenectomy were treated by transjugular approach catheter-directed thrombolysis. The mean age of the patients was 41.2 years. After access to the portal system via the transjugular approach, pigtail catheter fragmentation of clots, local urokinase injection, and manual aspiration thrombectomy were used for the initial treatment of PV-SMV thrombosis, followed by continuous thrombolytic therapy via an indwelling infusion catheter in the SMV, which was performed for three to six days. Adequate anticoagulation was given during treatment, throughout hospitalization, and after discharge. RESULTS: Technical success was achieved in all 6 patients. Clinical improvement was seen in these patients within 12-24 h of the procedure. No complications were observed. The 6 patients were discharged 6 14 d (8 +/- 2.5 d) after admission. The mean duration of follow-up after hospital discharge was 40 +/- 16.5 mo. Ultrasound and contrast-enhanced computed tomography confirmed patency of the PV and SMV, and no recurrent episodes of PV SMV thrombosis developed during the follow-up period. CONCLUSION: Catheter directed thrombolysis via transjugular intrahepatic access is a safe and effective therapy for the management of patients with symptomatic acute extensive PV-SMV thrombosis. PMID- 19554659 TI - Comparative identification of Ca2+ channel expression in INS-1 and rat pancreatic beta cells. AB - AIM: To identify and compare the profile of Ca(2+) channel subunit expression in INS-1 and rat pancreatic beta cells. METHODS: The rat insulin-secreting INS-1 cell line was cultured in RPMI-1640 with Wistar rats employed as islet donors. Ca(2+) channel subunit expression in INS-1 and isolated rat beta cells were examined by reverse transcription polymerase chain reaction (RT-PCR). Absolute real-time quantitative PCR was performed in a Bio-Rad iQ5 Gradient Real Time PCR system and the data analyzed using an iQ5 system to identify the expression level of the Ca(2+) channel subunits. RESULTS: In INS-1 cells, the L-type Ca(2+) channel 1C subunit had the highest expression level and the TPRM2 subunit had the second highest expression. In rat beta cells, the TPRC4beta subunit expression was dominant and the expression of the L-type 1C subunit exceeded the 1D subunit expression about two-fold. This result agreed with other studies, confirming the important role of the L-type 1C subunit in insulin-secreting cells, and suggested that non-voltage-operated Ca(2+) channels may have an important role in biphasic insulin secretion. CONCLUSION: Twelve major Ca(2+) channel subunit types were identified in INS-1 and rat beta cells and significant differences were observed in the expression of certain subunits between these cells. PMID- 19554660 TI - Measuring the space between vagina and rectum as it relates to rectocele. AB - AIM: To measure the normal space between the posterior wall of the vagina and the anterior wall of the respectively rectum using computed tomography (CT) and reveal its were relationship to rectocele. METHODS: A total of twenty female volunteers without rectocele were examined by CT scan. We performed a middle level continuous horizontal pelvic scan from the upper part to the lower part and collected the measurement data to analyze the results using t-test. RESULTS: Twenty volunteers were enrolled in the study. The space between the posterior wall of the vagina and the anterior wall of the rectum was measured at three levels (upper 1/3, middle, lower 1/3 level of vagina). The results showed that the space from the posterior wall of the vagina to the anterior wall of the rectum at the upper 1/3 level and the middle level was 3.896 +/- 0.3617 mm and 4.6575 +/- 0.3052 mm, respectively. When the two groups of data were compared, we found the space at the upper 1/3 level was shorter than at the middle level (P < 0.01). Moreover, at the lower 1/3 level the space measured was 10.058 +/- 0.4534 mm. The results revealed that the space at the lower 1/3 level was longer than that at the middle level (P < 0.01). CONCLUSION: These measurement data may be helpful in assessing rectocele clinical diagnosis and functional outcomes of rectocele repair. PMID- 19554661 TI - Current use of immunosuppressive agents in inflammatory bowel disease patients in East China. AB - AIM: To investigate immunosuppressive agents used to treat inflammatory bowel disease (IBD) in East China. METHODS: A retrospective review was conducted, involving 227 patients with IBD admitted to Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University from June 2000 to December 2007. Data regarding demographic, clinical characteristics and immunosuppressants usage were analyzed. RESULTS: A total of 227 eligible patients were evaluated in this study, including 104 patients with Crohn's disease and 123 with ulcerative colitis. Among the patients, 61 had indications for immunosuppressive agents use. However, only 21 (34.4%) received immunosuppressive agents. Among the 21 patients, 6 (37.5%) received a subtherapeutic dose of azathioprine with no attempt to increase the dosage. Of the 20 patients that received immunosuppressive agent treatment longer than 6 mo, 15 patients went into remission, four patients were not affected and one relapsed. Among these 20 patients, four patients suffered from myelotoxicity and one suffered from hepatotoxicity. CONCLUSION: Immunosuppressive agents are used less frequently to treat IBD patients from East China compared with Western countries. Monitoring immunosuppressive agent use is recommended to optimize dispensation of drugs for IBD in China. PMID- 19554662 TI - Hepatic injury induced by carbon dioxide pneumoperitoneum in experimental rats. AB - AIM: To observe the hepatic injury induced by carbon dioxide pneumoperitoneum in rats and to explore its potential mechanism. METHODS: Thirty healthy male SD rats were randomly divided into control group (n = 10), 0 h experimental group (n = 10) and 1 h experimental group (n = 10) after sham operation with carbon dioxide pneumoperitoneum. Histological changes in liver tissue were observed with hematoxylin-eosin staining. Liver function was assayed with an automatic biochemical analyzer. Concentration of malonyldialdehyde (MDA) and activity of superoxide dismutase (SOD) were assayed by colorimetry. Activity of adenine nucleotide translocator in liver tissue was detected with the atractyloside inhibitor stop technique. Expression of hypoxia inducible factor-1 (HIF-1) mRNA in liver tissue was detected with in situ hybridization. RESULTS: Carbon dioxide pneumoperitoneum for 60 min could induce liver injury in rats. Alanine aminotransferase and aspartate aminotransferase were 95.7 +/- 7.8 U/L and 86.8 +/ 6.9 U/L in 0 h experimental group, and 101.4 +/- 9.3 U/L and 106.6 +/- 8.7 U/L in 1 h experimental group. However, no significant difference was found in total billirubin, albumin, and pre-albumin in the three groups. In 0 h experimental group, the concentration of MDA was 9.83 +/- 2.53 micromol/g in liver homogenate and 7.64 +/- 2.19 micromol/g in serum respectively, the activity of SOD was 67.58 +/- 9.75 nu/mg in liver and 64.47 +/- 10.23 nu/mg in serum respectively. In 1 h experimental group, the concentration of MDA was 16.57 +/- 3.45 micromol/g in liver tissue and 12.49 +/- 4.21 micromol/g in serum respectively, the activity of SOD was 54.29 +/- 7.96 nu/mg in liver tissue and 56.31 +/- 9.85 nu/mg in serum, respectively. The activity of ANT in liver tissue was 9.52 +/- 1.56 in control group, 6.37 +/- 1.33 in 0 h experimental group and 7.28 +/- 1.45 (10(-9) mol/min per gram protein) in 1 h experimental group, respectively. The expression of HIF 1 mRNA in liver tissue was not detected in control group, and its optical density difference value was 6.14 +/- 1.03 in 0 h experimental group and 9.51 +/- 1.74 in 1 h experimental group, respectively. CONCLUSION: Carbon dioxide pneumoperitoneum during the sham operation can induce hepatic injury in rats. The probable mechanisms of liver injury include anoxia, ischemia reperfusion and oxidative stress. Liver injury should be avoided during clinical laparoscopic operation with carbon dioxide pneumoperitoneum. PMID- 19554663 TI - Palliative cardia resection with gastroesophageal reconstruction for perforated carcinoma of the gastroesophageal junction. AB - Iatrogenic perforation of esophageal cancer or cancer of the gastroesophageal (GE) junction is a serious complication that, in addition to short term morbidity and mortality, significantly compromises the success of any subsequent oncological therapy. Here, we present an 82-year-old man with iatrogenic perforation of adenocarcinoma of the GE junction. Immediate surgical intervention included palliative resection and GE reconstruction. In the case of iatrogenic tumor perforation, the primary goal should be adequate palliative (and not oncological) therapy. The different approaches for iatrogenic perforation, i.e. surgical versus endoscopic therapy are discussed. PMID- 19554664 TI - Development of an RPD CAD system with finite element stress analysis. AB - The structural design of removable partial dentures (RPDs) is critical for preventing distortion of the prosthesis, protecting abutment teeth and residual ridges as well as for high masticatory performance. The aim of this study was to clarify the feasibility and utility of a computer-aided designing (CAD) system with finite element analysis (FEA) for molar teeth arrangement in unilateral distal extension base RPDs. The shapes of artificial teeth and residual ridge were measured and converted into point group data. Solid models were created from surface-modelled point group data in a 3D surface CAD format. An occlusal rim was created on the residual ridge mucosa and the occlusal rim - residual ridge mucosa model with FEA function was created. Stress distribution on the residual ridge mucosa was compared by changing the loading point. The artificial teeth were then arranged in locations with the lowest amount of stress. After building an artificial teeth - saddle - residual ridge mucosa model, stress distribution in the residual ridge mucosa was re-evaluated by simulating occlusal force. On the occlusal rim - residual ridge mucosa model, stress was reduced when the loading point was located around the buccal shelf where functional cusps of artificial teeth were charted. It was confirmed that stress distribution in the residual ridge mucosa was equalized on the artificial teeth - saddle - residual ridge mucosa model. This system might be clinically useful tool for designing RPDs if FEA-guided designing of retainers and connectors can be added. PMID- 19554665 TI - Hormone-related factors and gynecological conditions in relation to endometrial cancer risk. AB - The objective of this study is to investigate the effect of menstrual and reproductive variables, breastfeeding, exogenous hormones, and gynecological conditions on endometrial cancer risk. We conducted a case-control study in Italy, including 454 women with endometrial cancer and 908 hospital controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using multivariate logistic regression models. Endometrial cancer risk was inversely associated with age at menarche (OR = 0.7, 95% CI = 0.5-1.0, for > or =14 vs. <12 years), and directly associated with age at menopause (OR = 1.8, 95% CI = 1.1 2.7, for > or =55 vs. <50 years) and years of menstruation (OR = 2.4, 95% CI = 1.7-3.4, for highest vs. lowest tertile). Multiparity strongly reduced the risk among women under 60 years of age (OR = 0.3, 95% CI = 0.2-0.6, for > or =3 deliveries vs. <2). Oral contraceptive use conferred a 40% reduced risk (95% CI = 0.4-1.0), irrespective of time since cessation. Although based on small numbers, women with a history of treated infertility (OR = 2.7, 95% CI = 1.1-6.4) or endometriosis (OR = 4.0, 95% CI = 1.0-15.5) were at increased risks. No significant associations with endometrial cancer risk emerged for age at first/last birth, breastfeeding, menopausal status, hormone replacement therapy, and history of uterine fibromyomas or polycystic ovary. In conclusion, this study confirms the importance of multiparity, years of menstruation, and oral contraceptive use in endometrial cancer etiology, thus contributing to identify women at elevated risk of such neoplasm. PMID- 19554666 TI - Contrast-enhanced whole-heart MR coronary angiography at 3.0 T using the intravascular contrast agent gadofosveset. AB - OBJECTIVES: The purpose of this study was to compare contrast-enhanced (CE) whole heart coronary magnetic resonance angiography (MRA) at 3.0 T using gadofosveset to noncontrast-enhanced steady-state free precession (SSFP) coronary MRA at 1.5 T. MATERIALS AND METHODS: A prospective randomized study was conducted among 20 healthy male volunteers. The same group of subjects underwent CE whole heart MRA at 3.0 T employing a 3D FLASH sequence with IR prepulse after gadofosveset injection as well as noncontrast-enhanced coronary MRA at 1.5 T using a 3D SSFP sequence with T2-preparation. Both techniques were performed using prospective ECG-triggering and adaptive respiratory gating. Acquisition time, signal-to-noise ratio of coronary blood, contrast-to-noise ratio (CNR) between coronaries and adjacent myocardium or epicardial fat, and image quality were evaluated in each case. RESULTS: A significant increase of the overall CNR between coronary blood and adjacent myocardium was measured on images acquired at 3 T in comparison to 1.5 T. The mean values were 38.9 +/- 19.6 and 26.3 +/- 15.4, respectively (P[r] < 0.005). There was no significant difference in CNR between coronary blood and epicardial fat. The mean image quality for the proximal and mid coronary segments was not statistically different between 1.5 T and 3.0 T (P > 0.05), however, the distal coronary segments were rated significantly higher for the CE MRA at 3.0 T (P = 0.02). The average acquisition time (15.29 +/- 5.73 minutes at 1.5 T vs. 17.29 +/- 5.18 minutes at 3 T) and overall image quality (2.15 +/- 0.49 at 1.5 T vs. 2.35 +/- 0.39 at 3 T) were similar for both methods. CONCLUSIONS: CE whole heart coronary MRA at 3.0 T demonstrated higher overall CNR between coronary blood and myocardium and an improved image quality of the distal coronary segments compared with noncontrast-enhanced SSFP coronary MRA at 1.5 T. PMID- 19554667 TI - In vivo detection of inflammation using pegylated iron oxide particles targeted at E-selectin: a multimodal approach using MR imaging and EPR spectroscopy. AB - OBJECTIVES: Ultrasmall particles of iron oxide (USPIO) possess superparamagnetic properties and are used as negative contrast agent in magnetic resonance imaging (MRI) because of their strong T(2) and T(2)* effects. Besides this method, electron paramagnetic resonance (EPR) offers the unique capability to quantify these particles. The objective of this study was to evaluate a molecular marker for non invasive diagnosis and monitoring of inflammation. During inflammation cell adhesion molecules such as E-selectin are expressed on the endothelial cell surface. An E-selectin ligand was coupled to pegylated USPIO particles. MATERIALS AND METHODS: Inflammation was induced by intramuscular injection of Freund's Complete Adjuvant in male NMRI mice. After intravenous injection of grafted or ungrafted USPIO particles, iron concentration in inflamed muscles was quantified ex vivo by X-band EPR. Particle accumulation was also assessed in vivo by L-Band EPR, as well as by T(2)-weighted MRI. RESULTS: We determined the mean iron oxide concentration in inflamed muscles after injection of grafted or ungrafted UPSIO particles, which was 0.8% and 0.4% of the initially injected dose, respectively. By L-band EPR, we observed that the concentration of the grafted USPIO particles in inflamed muscles was twice higher than for the ungrafted particles. Using MRI experiments, a higher signal loss was clearly observed in the inflamed muscle when grafted particles were injected in comparison with the ungrafted particles. CONCLUSION: Even taking into account a non specific accumulation of iron oxides, the targeting of USPIO particles with E-selectin ligands significantly improved the sensitivity of detection of inflamed tissues. PMID- 19554668 TI - Bupivacaine chondrotoxicity. PMID- 19554669 TI - [The sophism of the highest possible growth tables]. PMID- 19554670 TI - Practical depression screening in residential care/assisted living: five methods compared with gold standard diagnoses. AB - OBJECTIVE: To test the accuracy of five practical depression screening strategies in older adults residing in residential care/assisted living (RC/AL). DESIGN: Cross-sectional screening study. SETTING: Four RC/AL communities in North Carolina. PARTICIPANTS: A total of 112 residents aged > or =65 and 27 staff members involved in their care. MEASUREMENTS: Direct care staff was trained in and completed the Cornell Scale for Depression in Dementia, modified for use by long-term care staff (CSDD-M-LTCS). They additionally responded to a one-item question "Do you believe the resident is often sad or depressed?" and the Minimum Data Set Depression Rating Scale (DRS). Residents responded directly to the Geriatric Depression Scale (15-item version; GDS-15) and the Patient Health Questionnaire, 2-item version (PHQ-2). A geriatric psychiatrist performed gold standard diagnostic interviews using the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. Sensitivities and specificities were calculated for all instruments at predetermined cutpoints. RESULTS: Gold standard diagnoses yielded 14% prevalence of major or minor depression. The CSDD-M-LTCS and one-item screen completed by caregivers failed to significantly discriminate depressed cases. The DRS yielded high specificity (0.85) but low sensitivity (0.47). For the two resident reported measures, the PHQ-2 had a sensitivity of 0.80 and specificity of 0.71, and the GDS-15, 0.60 and 0.75, respectively. CONCLUSION: Measures completed by caregivers failed to adequately detect depression. Of the measures completed directly by residents, the PHQ-2 seems to have the best mix of brevity, sensitivity, and ease of administration. PMID- 19554671 TI - Comorbid depression in dementia on psychogeriatric nursing home wards: which symptoms are prominent? AB - OBJECTIVE: To provide insight into the prevalence and clinically relevant symptoms of comorbid depression among dementia patients in psychogeriatric nursing home wards, to enhance depression recognition. DESIGN: Cross-sectional analyses of multicenter diagnostic data. SETTING: Psychogeriatric wards of Dutch nursing homes. PARTICIPANTS: Five hundred and eighteen residents with dementia. MEASUREMENTS: 1) Diagnosis of depression in dementia (Provisional Diagnostic Criteria for Depression of Alzheimer disease [PDC-dAD]), 2) dementia (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition-PC), and 3) stage of dementia (Geriatric Depression Scale). RESULTS: The point prevalence of comorbid depression in dementia (Stages 2-6) on psychogeriatric nursing home wards was 19%. "Depressed mood," "irritability," and "fatigue" were the most prevalent depressive symptoms. Residents taking antidepressants at the time of the PDC-dAD depression diagnosis showed more depressive symptoms than residents who were not. The mean number of depressive symptoms was 5.6 (SD 1.84), which did not differ between the dementia stages. Also, no differences were found in the point prevalence of the shown symptoms between dementia stages. CONCLUSION: Irritability was put forward by the developers of the PDC-dAD, as one of the specific symptoms of depression in Alzheimer disease. This study shows that irritability is one of the most prevalent depressive symptoms in psychogeriatric nursing home residents diagnosed with comorbid depression. Irritability should therefore alert caregivers to the presence of depression and could help early recognition. The high-prevalence rate of comorbid depression in dementia in this setting justifies attention to early recognition and intervention. PMID- 19554672 TI - Variability in depressive symptoms predicts cognitive decline in age-related macular degeneration. AB - OBJECTIVE: The measurement of affective symptoms in older persons who decline cognitively is uncertain. The authors investigated whether mood variability predicts dementia in patients with age-related macular degeneration (AMD). DESIGN: Three-year observational study after a clinical trial. SETTING: Community follow-up of outpatients ascertained from retina clinics. PARTICIPANTS: One hundred sixty patients with AMD. MEASUREMENTS: Geriatric Depression Scale (GDS) administered every 2 weeks for 6 months to subjects; Informant Questionnaire for Cognitive Decline in the Elderly (IQCODE) administered to subjects' knowledgeable informants. RESULTS: Twenty-three subjects (14.4%) declined cognitively. Age, education, baseline GDS score > or =5, and variability in GDS scores (i.e., fluctuations between adjacent time points) were associated with cognitive decline. For GDS variability, each 1 unit increase in the residual standard deviation (SD) of the GDS increased the risk for cognitive decline by 93% (IDR = 1.92; 95% CI [1.27-2.91]). Thus, subjects with a residual SD of 1 were nearly twice as likely to become demented as subjects with no variability in GDS scores. The risk for subjects with SDs of 2 increased more than threefold (IDR = 3.68; 95% CI [1.61-8.47]). A multiple regression analysis showed that GDS variability was a significant risk factor for dementia after controlling for significant covariates. CONCLUSION: These data suggest a useful approach to conceptualizing and measuring depressive symptoms in older persons. Variability in self-reported mood may be an early sign of dementia and may offer new insights into the neurobiological mechanisms linking depression and cognition PMID- 19554674 TI - Modeling the connectivity of the neural systems. PMID- 19554673 TI - The personality domains and styles of the five-factor model are related to incident depression in Medicare recipients aged 65 to 100. AB - OBJECTIVES: Few prospective studies have examined personality and depression in older adults. The authors investigated whether the Five-Factor Model of personality traits-Neuroticism, Extraversion, Openness to Experience, Agreeableness, and Conscientiousness-and trait combinations (styles) are related to incident major or minor depression. PARTICIPANTS/SETTING: Prospective data were gathered on a community sample of 512 older adults with disability and a history of significant health care utilization who were enrolled in a Medicare Demonstration Project. MEASUREMENTS: Depression and personality traits and styles were assessed at baseline; depression was assessed again at approximately 12 and 22 months. DESIGN: Participants who developed incident major depression were compared with those free of depression at all three assessments. Similar analyses were done for minor depression. RESULTS: High Neuroticism and low Conscientiousness were risk factors for both major and minor depression. Combinations of high Neuroticism with low or high Extraversion or high Openness conferred risk for major depression. Other novel findings for major depression revealed new trait combinations of low Conscientiousness with low or high Extraversion, high Openness, or low Agreeableness. Three trait combinations, all involving low Conscientiousness, predicted risk for minor depression: high Neuroticism, high Agreeableness, and low Openness. CONCLUSION: The present findings highlight the importance of examining combinations of personality traits or personality styles when identifying those who are most at-risk for geriatric depression. Since other personality domains may modify the risk related to high Neuroticism and low Conscientiousness, the prevention, diagnosis, and treatment of depression could be greatly improved by assessing older patients not only on all five domains of personality but in terms of their combinations. PMID- 19554675 TI - Long term use of topical tacrolimus (FK506) in high-risk penetrating keratoplasty. PMID- 19554676 TI - [Prevalence and clinical significance of malnutrition and effectiveness of nutritional support for patients suffering from chronic heart failure]. AB - This multicenter study evaluated the effectiveness of threpsology support during the progression of chronic heart failure (HSN). The paper shows the preliminary results of a study SATURN II. We show that nutritivnaya support is effective in any degree ofmalnutrition in patients with HSN. PMID- 19554677 TI - [Actual problems of helicobacteriosis diagnostics]. AB - In this article the comparative analysis of various methods of Helicobacter pylori verification is shown. Also algorithm of optimization of helicobacteriosis diagnostic, developed on the basis of a comparative estimation of results of various diagnostic techniques is presented. According to this algorithm, for increase of accuracy of helicobacteriosis diagnostic it is recommended to use at least two, and it is better three diagnostic methods: the combination of fast urease test or "Helic-test" with a histologic method (biopsy material from a stomach corpus) or polymerase chain reaction (ureC gene detection) is preferable. For an efficiency estimation of eradication therapy we recommend "Helic-test" as an effective noninvasive method that is especially important for children. PMID- 19554678 TI - Redox-coupled complexation of 23-phospha-21-thiaporphyrin with group 10 metals: a convenient access to stable core-modified isophlorin-metal complexes. PMID- 19554679 TI - Bamboo-type TiO2 nanotubes: improved conversion efficiency in dye-sensitized solar cells. PMID- 19554680 TI - Selective oxidation of ethanol to acetaldehyde on gold. PMID- 19554681 TI - One site is enough. Catalytic water oxidation by [Ru(tpy)(bpm)(OH2)]2+ and [Ru(tpy)(bpz)(OH2)]2+. PMID- 19554682 TI - In situ monitoring of bindings between dasatinib and its target protein kinases using magnetic nanoparticles in live cells. PMID- 19554683 TI - Stable lariat formation based on a G-quadruplex scaffold. PMID- 19554684 TI - Indole synthesis via rhodium catalyzed oxidative coupling of acetanilides and internal alkynes. PMID- 19554685 TI - Synthesis and reactivity of functionalized arylcopper compounds by transmetalation of organosilanes. PMID- 19554686 TI - A ruthenium-catalyzed, atom-economical synthesis of nitrogen heterocycles. PMID- 19554687 TI - Initial catalyst-substrate association step in enyne metathesis catalyzed by grubbs ruthenium complex probed by time-dependent fluorescence quenching. PMID- 19554688 TI - Using a sterically restrictive amino acid as a 19F NMR label to monitor and to control peptide aggregation in membranes. PMID- 19554689 TI - An additional spirocyclization for duocarmycin SA. PMID- 19554690 TI - Controlled assembly of bifunctional chimeric protein cages and composition analysis using noncovalent mass spectrometry. PMID- 19554691 TI - Links between single-site heterogeneous and homogeneous catalysis. DFT analysis of pathways for organozirconium catalyst chemisorptive activation and olefin polymerization on gamma-alumina. AB - This contribution focuses on the catalytic properties of the organozirconium precatalyst Cp2Zr(CH3)2 chemisorbed on dehydroxylated gamma-alumina (Al2O3), as analyzed via density functional theory. The interactions of the catalytically active cationic Cp2ZrCH3+ adsorbate species are scrutinized at two possible model Al2O3 (110) surface sites, namely mu2-O and mu3-O, representing the principal reactive species on the alumina surface. It is found that zirconocenium coordination occurs via two different geometries (dioxo-bridged and oxo-bridged) at both the mu-O and mu2-O surface sites. This process is compared to that for forming the related homogeneous phase Cp2ZrCH3+H3CB(CeFs)3- ion pair structure. It is found that the interaction of the Cp2ZrCH3 adsorbate species with the Pr2-O sites is far stronger than that with the B3-O sites due to the greater unsaturation of the former. Furthermore, the interaction with the mu3-O sites is weaker than that in the parent homogeneous ion pair. The catalytic activity of the chemisorbed Cp2ZrCH3+ systems for ethylene polymerization is investigated at both mu2-O and mu3-O sites and compared with the analogous Cp2ZrCH3+H3CB(C6F5)3- mediated process in solution. A Cossee enchainment mechanism proceeds via ethylene pi-complex formation and an alpha-agostic assisted transition state to yield gamma- and beta-agostic insertion products. The overall kinetics of enchainment are closely correlated with the energetics of pi-complex formation, and it is suggested that the differing kinetic behaviors of the surface-bound Cp2ZrR+ species on the various Al2O3 coordination sites and the analogous homogeneous species reflect differences in the olefin pi-complex stabilization energies. These computational results agree well with the experimental data which indicate that only fractions of the surface bound species are catalytically significant but that these are far more catalytically active than the homogeneous analogues. PMID- 19554692 TI - Electronic effects on the surface potential at the vapor-liquid interface of water. AB - The surface potential of the vapor-liquid interface of pure water is relevant to electrochemistry, solvation thermodynamics of ions, and interfacial reactivity. The chemistry of an ion near the vapor-liquid interface is influenced by the surface potential. Indirect determinations of the surface potential have been experimentally attempted many times, yet there has been little agreement as to its magnitude and sign (-1.1 to +0.5 V). We present the first computation of the surface potential of water using ab initio molecular dynamics and find a surface potential of -18 mV with a maximum interfacial electric field of +8.9 x 10(7) V/m, which are consistent with structural data from experiment. A comparison is made between our results and those from experiments and previous molecular simulations. The associated electric field can alter interfacial reactivity and transport, while the surface potential can be used to determine the "chemical" contribution to the real and electrochemical potentials for ion transport through the vapor-liquid interface. PMID- 19554693 TI - Observing metal-catalyzed chemical reactions in situ using surface-enhanced Raman spectroscopy on Pd-Au nanoshells. AB - Insight into the nature of transient reaction intermediates and mechanistic pathways involved in heterogeneously catalyzed chemical reactions is obtainable from a number of surface spectroscopic techniques. Carrying out these investigations under actual reaction conditions is preferred but remains challenging, especially for catalytic reactions that occur in water. Here, we report the direct spectroscopic study of the catalytic hydrodechlorination of 1,1 dichloroethene in H2O using surface-enhanced Raman spectroscopy (SERS). With Pd islands grown on Au nanoshell films, this reaction can be followed in situ using SERS, exploiting the high enhancements and large active area of Au nanoshell SERS substrates, the transparency of Raman spectroscopy to aqueous solvents, and the catalytic activity enhancement of Pd by the underlying Au metal. The formation and subsequent transformation of several adsorbate species was observed. These results provide the first direct evidence of the room-temperature catalytic hydrodechlorination of a chlorinated solvent, a potentially important pathway for groundwater cleanup, as a sequence of dechlorination and hydrogenation steps. More broadly, the results highlight the exciting prospects of studying catalytic processes in water in situ, like those involved in biomass conversion and proton exchange membrane fuel cells. PMID- 19554694 TI - Van der Waals versus hydrogen-bonding forces in a crystalline analog of cellotetraose: cyclohexyl 4'-O-cyclohexyl beta-D-cellobioside cyclohexane solvate. AB - Hydrogen bonding is important in cellulosic and other carbohydrate structures, but the role of interactions between nonpolar groups is less understood. Therefore, we synthesized cyclohexyl 4'-O cyclohexyl beta-D-cellobioside (8), a molecule that has two glucose rings and two nonpolar cyclohexyl rings. Key to attaching the 4'-Ocyclohexyl group was making the 4'-O,6'-O-cyclohexylidene ketal. After peracetylation, the cyclohexylidene ketal ring was opened regioselectively, providing 65% of 8 after final deacetylation. Comparison of the crystal structure of 8, as the cyclohexane solvate, with those of cellulose and its fragments, especially cellotetraose with four glucose rings, revealed extensive effects from the cyclohexyl groups. Three conformationally unique molecules (A, B, and C) are in the triclinic unit cell of 8, along with two solvent cyclohexanes. When viewed down the crystal's a-axis, the array of C, A, and B looks like the letter N, with A inclined so that its cyclohexyl groups can stack with those of the reducing ends of the B and C molecules. The lower left and upper right points of the N are stacks of cyclohexyl rings on the nonreducing ends of B and C, interspersed with solvent cyclohexanes. Whereas cellotetraose has antiparallel (up-down) packing, A and B in 8 are oriented "down" in the unit cell while C is "up". "Down-down-up" (or, alternatively, "up-up-down") packing is rare for carbohydrates. Other unusual details include 06 in all three staggered orientations: one is tg, two are gg, and three are gt, confirmed with CP/MAS 13C NMR. The tg O6 donates a proton to an intramolecular hydrogen bond to O2', opposite to the major schemes in native cellulose I. A similar but novel O6B H...O2'B hydrogen bond is based on a slightly distorted gg orientation. The hydrogen bonds between parallel molecules are unique, with linkages between O2'A and O2'B, O3'A and O3'B, and O6A and O6B. Other details, such as the bifurcated O3...O5' and ...O6' hydrogen bonds are similar to those of other cellulosic structures. C-H...O hydrogen bonds are extensive along the [110] line of quarter staggering. The unusual features described here expand the range of structural motifs to be considered for as-yet undetermined cellulose structures. PMID- 19554695 TI - Biomimetic cleavage of RNA models promoted by a dinuclear Zn(II) complex in ethanol. Greater than 30 kcal/mol stabilization of the transition state for cleavage of a phosphate diester. AB - The cleavage of a series of seven substituted aryl 2-hydroxypropyl phosphates (1a g) promoted by a dinuclear Zn(II) complex (3:Zn(II)2:(-OCH2CH3)) was investigated in ethanol at pH 9.0 +/- 0.2 and 25 degrees C. The kinetics for appearance of the product phenols follow very strong saturation behavior for all substrates where the dissociation constant of the bound complex has an upper limit of Km = 3 x 10( 7) M and the k(cat)(max corr.) values (corrected for triflate inhibition) range from 168 to 3 s(-1). A partial s(s)pH/log k(cat)(max corr). profile for the 3:Zn(II)2:(-OCH2CH3)-catalyzed reaction of le (3-methoxyphenyl 2-hydroxypropyl phosphate) is bell-shaped, plateauing from 7.9-10, and is fit to a two kinetically important ionizations having s(s)pKa values of 7.22 and 10.9. The Bronsted plot of log (k(cat)(max corr.)) vs. the s(s)pKa values for the phenols shows a break at about 14.3 with two beta(lg), values of -1.12 and 0.0. This is analyzed in terms of a change in rate limiting step from cleavage of the phosphate to a conformational change where the binding of the phosphate changes from one P-O- ----Zn(II) interaction to a Zn(II)----O-P-O---Zn(II) double activation. An energetics calculation comparing the ethoxide promoted cleavage of 1a-g with the 3:Zn(ll)2:(-OEt) promoted reaction indicates that the complex, 3:Zn(II)2, stabilizes the ethoxide plus substrate transition state for the cleavage of 1a-g by between 33 and 36 kcal/mol. The origins of the large stabilization are discussed in terms of the effect of the medium on the various rate and equilibrium constants involved. PMID- 19554696 TI - Group 10 and 11 metal boratranes (Ni, Pd, Pt, CuCl, AgCl, AuCl, and Au+) derived from a triphosphine-borane. AB - The ambiphilic triphosphine-borane ligand 1 {TPB = [o-iPr2P-(C6H4)3B} readily coordinates to all group 10 and 11 metals to afford a complete series of metal boratranes (TPB)[M] 2-8 (2: M = Ni, 3: M = Pd, 4: M = Pt, 5: M = CuCl, 6: M = AgCl, 7: M = AuCl, 8: M = Au+). Spectroscopic and structural characterization unambiguously establishes the presence of M-B interactions in all of these complexes. The first evidence for borane coordination to copper and silver is provided, and the Au-->B interaction is shown to persist upon chloride abstraction. Experimental and theoretical considerations indicate that the M-->B interaction is strongest in the Pt and Au complexes. The influence of the oxidation state and charge of the metal is substantiated, and the consequences of relativistic effects are discussed. The coordination of the sigma-acceptor borane ligand is found to induce a significant bathochromic shift of the UV-vis spectra, the Ni, Pd, and Pt complex presenting strong absorptions in the visible range. In addition, all of the group 10 and 11 metal boratranes adopt C3 symmetry both in the solid state and in solution. The central M-->B interaction is found to moderately influence the degree of helicity and configurational stability of these three-bladed propellers, and DFT calculations support a dissociative pathway for the inversion process. PMID- 19554697 TI - Dendritic molecular switch: chiral folding and helicity inversion. AB - Appropriately designed chemical architectures can fold to adopt well-defined secondary structures without the need for structural motifs of biological origin. We have designed tris(N-salicylideneaniline)-based hyperbranched molecules that spontaneously collapse to compact three-blade propeller geometry of either (P)- or (M)-handedness. For a homologous series of compounds, a direct correlation was established between the absolute screw sense, either (P)- or (M)-, of this helical folding and the absolute configuration, either (R)- or (S)-, of the chiral alcohol groups introducing local asymmetric bias to the conformationally restricted molecular backbone. 1H NMR and CD spectroscopic studies provided significant insights into structural folding and unfolding of these chiral molecules in solution, which proceed via reversible assembly and disassembly of the C3-symmetric hydrogen-bonding network. Notably, solvents profoundly influenced this dynamic process. A strong correlation between the solvent donor number (DN) or solvent basicity (SB) parameters and the change in the Cotton effects pointed toward specific O-H...solvent interactions that drive structural unfolding and eventual refolding to apparently opposite helicity. This unusual chirality inversion process could also be induced by installation of chemical protecting groups that simulate specific solvent-solute interactions. Removal of this covalent mimic of the solvent shell restored the original screw sense of the parent molecule, thus establishing the feasibility of covalently triggered helicity inversion as a new mode of operation for chiroptical molecular switches. PMID- 19554698 TI - Palliative care in rural areas. PMID- 19554699 TI - Falling into snobbery traps. PMID- 19554700 TI - Paperwork duplication. PMID- 19554701 TI - Support for skin cancer nurses. PMID- 19554702 TI - Shocked at fee increase. PMID- 19554703 TI - Fairness in English language testing. PMID- 19554705 TI - Photo brings back memories. PMID- 19554704 TI - Help needed with blood pressure cuffs. PMID- 19554706 TI - Getting the most from nursing. PMID- 19554707 TI - Students should take responsibility. PMID- 19554708 TI - The hype over herceptin. PMID- 19554709 TI - Speaking truth. PMID- 19554711 TI - Clinical effectiveness of photodynamic therapy in the treatment of periodontitis. AB - AIM: A randomized-controlled clinical pilot trial was designed to evaluate photodynamic therapy (PDT) for its bactericidal potential and clinical effect in the treatment of periodontitis. MATERIAL AND METHODS: Fifty-eight subjects with chronic periodontitis were included. Each subject exhibited at least three active periodontal pockets 5mm or deeper, bleeding on probing and the presence of Porphyromonas gingivalis. Subjects were randomly assigned to a control group treated by subgingival ultrasound only or to a study group additionally treated by PDT. Baseline clinical values of gingival index, bleeding on probing, probing pocket depths and clinical attachment levels were recorded and re-evaluated 90 days later. Pathogen screening for P. gingivalis, Tannerella forsythia and Treponema denticola was conducted at baseline as well as 10, 42 and 90 days after treatment. RESULTS: P. gingivalis was significantly reduced in both groups (laser group: p=0.020; control group: p=0.042). No significant reductions of T. forsythia and T. denticola were observed in either group. For the microbial parameters, no significant difference was found between the laser and the control group. All clinical parameters were significantly reduced in both groups after treatment. The mean probing pocket depths decreased from 5.79 to 4.55mm in the laser group and from 5.54 to 4.51 in the control group. The intergroup difference was not significant (p=0.82). Bleeding on probing was reduced from 100% evaluated at baseline to 47% in the laser group and 59% in the control group. The intergroup difference was not significant (p=0.28). No significant differences were observed in any other parameters. CONCLUSION: Application of a single cycle of PDT was not effective as an adjunct to ultrasonic periodontal treatment. There were no extra reductions in pocket depths and bleeding on probing. With regard to eradicating bacteria, however, there are no additional effects as compared with conventional treatment alone. PMID- 19554712 TI - Final manuscript in the Food Allergy educational series: diagnosis of immunoglobulin E-mediated food allergy. PMID- 19554713 TI - Epigenetic regulation of nervous system development by DNA methylation and histone deacetylation. AB - Alterations in the epigenetic modulation of gene expression have been implicated in several developmental disorders, cancer, and recently, in a variety of mental retardation and complex psychiatric disorders. A great deal of effort is now being focused on why the nervous system may be susceptible to shifts in activity of epigenetic modifiers. The answer may simply be that the mammalian nervous system must first produce the most complex degree of developmental patterning in biology and hardwire cells functionally in place postnatally, while still allowing for significant plasticity in order for the brain to respond to a rapidly changing environment. DNA methylation and histone deacetylation are two major epigenetic modifications that contribute to the stability of gene expression states. Perturbing DNA methylation, or disrupting the downstream response to DNA methylation - methyl-CpG-binding domain proteins (MBDs) and histone deacetylases (HDACs) - by genetic or pharmacological means, has revealed a critical requirement for epigenetic regulation in brain development, learning, and mature nervous system stability, and has identified the first distinct gene sets that are epigenetically regulated within the nervous system. Epigenetically modifying chromatin structure in response to different stimuli appears to be an ideal mechanism to generate continuous cellular diversity and coordinate shifts in gene expression at successive stages of brain development - all the way from deciding which kind of a neuron to generate, through to how many synapses a neuron can support. Here, we review the evidence supporting a role for DNA methylation and histone deacetylation in nervous system development and mature function, and present a basis from which to understand how the clinical use of HDAC inhibitors may impact nervous system function. PMID- 19554714 TI - Formation of a bridged butterfly-type mu-eta2:eta2-peroxo dicopper core structure with a carboxylate group. PMID- 19554715 TI - Selective ion passage through functionalized graphene nanopores. PMID- 19554716 TI - Sensitivity and multiplexing capabilities of MRI based on polarized 129Xe biosensors. PMID- 19554717 TI - Catalytic enantioselective additions of indoles to nitroalkenes. PMID- 19554718 TI - 6'-derivatised alpha-GalCer analogues capable of inducing strong CD1d-mediated Th1-biased NKT cell responses in mice. PMID- 19554719 TI - Oxidation state-specific fluorescent method for palladium(II) and platinum(IV) based on the catalyzed aromatic claisen rearrangement. PMID- 19554720 TI - A catalytic cycle for oxidation of tert-butyl methyl ether by a double C-H activation-group transfer process. PMID- 19554721 TI - Direct imaging of the ligand monolayer on an anion-protected metal nanoparticle through cryogenic trapping of its solution-state structure. PMID- 19554723 TI - Gold-catalyzed 1,3-addition of a sp3-hybridized C-H bond to alkenylcarbenoid intermediate. PMID- 19554722 TI - Self-assembled dinuclear cobalt(II)-salen catalyst through hydrogen-bonding and its application to enantioselective nitro-aldol (Henry) reaction. PMID- 19554724 TI - Highly enantioselective carbonyl-ene reactions catalyzed by In(III)-PyBox complex. PMID- 19554725 TI - Quasi 2D colloidal CdSe platelets with thicknesses controlled at the atomic level. PMID- 19554726 TI - An "inside-out" microfluidic approach to monodisperse emulsions stabilized by solid particles. PMID- 19554728 TI - Mechanistic studies of the hydroamination of norbornene with electrophilic platinum complexes: the role of proton transfer. AB - Hydroaminations of norbornene with arylsulfonamides and weakly basic anilines were achieved using electrophilic Pt(II) bis(triflate) complexes of the type L2Pt(OTf)2 (L2 = (t)Bu2bpy, (t)BuC6H4N== C(CH3)C(CH3)==NC6H4(t)Bu, (C6H5)2PCH2CH2P(C6H5)2, (C6F5)2PCH2CH2P(C6F5)2, S-BINAP). Pseudo-first-order kinetics reveal little to no dependence of the reaction rate on the ancillary ligand. Mechanistic studies do not favor an olefin coordination mechanism but are instead consistent with a mechanism involving sulfonamide coordination and generation of an acidic proton that is transferred to the norbornene. It is postulated that the resulting norbornyl cation is then attacked by free sulfonamide, and loss of proton from this adduct completes the hydroamination. The platinum-sulfonamide complex readily undergoes deprotonation to give a mu amido platinum-bridged dimer that was isolated from the reaction solution. These studies also involve use of Me3SiPh and Me3SnPh as non-nucleophilic proton traps. Cleavage of the Ph-E bonds was used to detect the acidic, catalytically active species. PMID- 19554727 TI - Efficient catalytic promiscuity in an enzyme superfamily: an arylsulfatase shows a rate acceleration of 10(13) for phosphate monoester hydrolysis. AB - We report a second catalytic activity of Pseudomonas aeruginosa arylsulfatase (PAS). Besides hydrolyzing sulfate monoesters, this enzyme catalyzes the hydrolysis of phosphate monoesters with multiple turnovers (>90), a k(cat) value of 0.023 s(-1), a K(M) value of 29 microM, and a kcat/K(M) ratio of 790 M(-1) s( 1) at pH 8.0. This corresponds to a remarkably high rate acceleration of 10(13) relative to the nonenzymatic hydrolysis [(k(cat)/K(M))/k(w)] and a transition state binding constant (K(tx)) of 3.4 pM. Promiscuous phosphatase and original sulfatase activities only differ by a factor of 620 (measured by k(cat)), so the enzyme provides high accelerations for both reactions. The magnitudes and relative similarity of the kinetic parameters suggest that a functional switch from sulfatase to phosphatase activities is feasible, either by gene duplication or by direct evolution via an intermediate enzyme with dual specificity. PMID- 19554730 TI - Nucleation and growth of ZnO in organic solvents--an in situ study. AB - ZnO is a metal oxide material which possesses versatile properties and applications. Therefore, the target-oriented preparation of ZnO has become a major issue. Many preparation techniques involve bottom-up methods from precursor solutions. In the current contribution, a special precursor system is described that enables a fine-control of kinetic parameters for the nucleation and growth of ZnO in various organic solvents. A large variety of analytical techniques could be applied in an in situ fashion to probe for the ZnO formation at all times and all length scales. Among the analytical techniques are UV/vis, Raman, Fluorescence, X-ray absorption, 1H NMR-spectroscopy, dynamic light-scattering, and TEM. Three different regimes for nucleation and growth with different characteristics could be identified. Furthermore, the effect of different parameters on the resulting ZnO particle size was investigated. PMID- 19554729 TI - Nanoscale growth factor patterns by immobilization on a heparin-mimicking polymer. AB - In this study, electrostatic interactions between sulfonate groups of an immobilized polymer and the heparin binding domains of growth factors important in cell signaling were exploited to nanopattern the proteins. Poly(sodium 4 styrenesulfonate-co-poly(ethylene glycol) methacrylate) (pSS-co-pPEGMA) was synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization using ethyl S-thiobenzoyl-2-thiopropionate as a chain transfer agent and 2,2'-azoisobutyronitrile (AIBN) as the initiator. The resulting polymer (1) was characterized by 1H NMR, GPC, FT-IR, and UV-vis and had a number average molecular weight (Mn) of 24,000 and a polydispersity index (PDI) of 1.17. The dithioester end group of 1 was reduced to the thiol, and the polymer was subsequently immobilized on a gold substrate. Binding of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) to the polymer via the heparin binding domains was then confirmed by surface plasmon resonance (SPR). The interactions were stable at physiological salt concentrations. Polymer 1 was cross-linked onto silicon wafers using an electron beam writer forming micro- and nanopatterns. Resolutions of 100 nm and arbitrary nanoscale features such as concentric circles and contiguous squares and triangles were achieved. Fluorescence microscopy confirmed that bFGF and VEGF were subsequently immobilized to the polymer micro- and nanopatterns. PMID- 19554731 TI - Current transients in single nanoparticle collision events. AB - Electrochemical hydrazine oxidation and proton reduction occur at a significantly higher rate at Pt than at Au or C electrodes. Thus, the collision and adhesion of a Pt particle on a less active Au or C electrode leads to a large current amplification by electrocatalysis at single nanoparticles (NPs). At low particle concentrations, the collision of Pt NPs was characterized by current transients composed of individual current profiles that rapidly attained a steady state, signaling single NP collisions. The characteristic steady-state current was used to estimate the particle size. The fluctuation in collision frequency with time indicates that the collision of NPs at the detector electrodes occurs in a statistically random manner, with the average frequency a function of particle concentration and diffusion coefficient. A longer term current decay in single current transients, as opposed to the expected steady-state behavior, was more pronounced for proton reduction than for hydrazine oxidation, revealing microscopic details of the nature of the particle interaction with the detector electrode and the kinetics of electrocatalysis at single NPs. The study of single NP collisions allows one to screen particle size distributions and estimate NP concentrations and diffusion coefficients. PMID- 19554732 TI - Modeling differential binding of alpha4beta2 nicotinic acetylcholine receptor with agonists and antagonists. AB - Three-dimensional structures of both the open- and closed-channel states of alpha4beta2 receptor have been modeled and used to study their binding with representative agonists and antagonists. The obtained binding structures and free energies consistently reveal that antagonists bind more favorably with the closed channel state and agonists bind more favorably with the open-channel state. The computational insights have led us to propose a computational strategy and protocol predicting whether a receptor ligand is an agonist or antagonist. Using the computational protocol, one only needs to calculate the relative binding free energies for a ligand binding with the open- and closed-channel structures. The ligand is predicted to be an agonist if the binding free energy calculated for the ligand binding with the open-channel state is significantly lower than that for its binding with the closed-channel state. If the binding free energy of a ligand with the open-channel state is higher than that with the closed-channel, the ligand is predicted to be an antagonist. The binding free energies calculated for all of the ligands binding with their most favorable channel states of the receptor are all close to the corresponding experimentally derived binding free energies. The new computational insights obtained and novel computational strategy and protocol proposed in this study are expected to be valuable in structure-based rational design of novel agonists/antagonists of nAChRs as therapeutic agents. PMID- 19554733 TI - MoS2 nanoribbons: high stability and unusual electronic and magnetic properties. AB - First-principles computations were carried out to predict the stability and magnetic and electronic properties of MoS2 nanoribbons with either zigzag- or armchair-terminated edges. Zigzag nanoribbons show the ferromagnetic and metallic behavior, irrespective of the ribbon width and thickness. Armchair nanoribbons are nonmagnetic and semiconducting, and the band gaps converge to a constant value of approximately 0.56 eV as the ribbon width increases. The higher stability of MoS2 nanoribbons, compared with the experimentally available triangular MoS2 nanoclusters, invites the experimental realization of such novel ribbons in true nanoscale. PMID- 19554735 TI - Ryan White: an unintentional home builder. PMID- 19554734 TI - Functionalized single-walled carbon nanotubes as rationally designed vehicles for tumor-targeted drug delivery. AB - A novel single-walled carbon nanotube (SWNT)-based tumor-targeted drug delivery system (DDS) has been developed, which consists of a functionalized SWNT linked to tumor-targeting modules as well as prodrug modules. There are three key features of this nanoscale DDS: (a) use of functionalized SWNTs as a biocompatible platform for the delivery of therapeutic drugs or diagnostics, (b) conjugation of prodrug modules of an anticancer agent (taxoid with a cleavable linker) that is activated to its cytotoxic form inside the tumor cells upon internalization and in situ drug release, and (c) attachment of tumor-recognition modules (biotin and a spacer) to the nanotube surface. To prove the efficacy of this DDS, three fluorescent and fluorogenic molecular probes were designed, synthesized, characterized, and subjected to the analysis of the receptor mediated endocytosis and drug release inside the cancer cells (L1210FR leukemia cell line) by means of confocal fluorescence microscopy. The specificity and cytotoxicity of the conjugate have also been assessed and compared with L1210 and human noncancerous cell lines. Then, it has unambiguously been proven that this tumor-targeting DDS works exactly as designed and shows high potency toward specific cancer cell lines, thereby forming a solid foundation for further development. PMID- 19554736 TI - Establishing an anal dysplasia clinic for HIV-infected men: initial experience. AB - Anal dysplasia caused by human papillomavirus (HPV) infection is common in the HIV-infected population and is a precursor to squamous cell carcinoma of the anus (SCCA). Herein, we describe our initial experience in assessing the frequency and severity of anal intraepithelial neoplasia (AIN) in a newly formed anal dysplasia clinic in Seattle. During a 7-month period, 150 HIV-positive men were evaluated by digital rectal examination and high-resolution anoscopy (HRA). Forty-seven patients of the 122 patients who underwent biopsy (39%) had biopsy-identified low grade AIN, and 47 (39%) had high-grade AIN (HGAIN). Two patients with HGAIN were referred for surgical treatment and were further noted to have microinvasive SCCA. No patient reported significant post-HRA biopsy complications. Patient tolerance and acceptance of AIN screening was good, and the majority of those who underwent screening have been adherent to recommended follow-up examinations and treatment. The clinic is expanding rapidly, with excellent community and provider support. We anticipate that the anal dysplasia clinic will enable our institution to participate in emerging HIV- and HPV-related AIN clinical trials. PMID- 19554737 TI - Editorial comment: screening for anal dysplasia--are we on the same page? PMID- 19554738 TI - Images in HIV/AIDS. Keloid after herpes zoster in an HIV-infected person. PMID- 19554739 TI - Acute MI with long-standing AIDS: a perfect storm of risk factors for premature coronary artery disease. PMID- 19554740 TI - Expanding HIV testing: overcoming physician barriers. PMID- 19554741 TI - Policy watch. A tale of two cities. PMID- 19554743 TI - The health benefits of tai chi. This gentle form of exercise can prevent or ease many ills of aging and could be the perfect activity for the rest of your life. PMID- 19554742 TI - Experts recommend low-dose aspirin to prevent stroke in women. Lower doses are as effective as higher doses and are likely to be safer. PMID- 19554744 TI - What to do about an ingrown toenail. Warm soaks and proper nail trimming may do the trick, but sometimes it's best to see a foot specialist right away. PMID- 19554745 TI - High resting heart rate predicts heart risk in women at midlife. PMID- 19554746 TI - Exercise eases nighttime leg twitches. PMID- 19554747 TI - By the way, doctor. I'm 69 and have taking Fosamax for 10 years. My doctor has approved a "drug holiday," provided we use a CTX test to monitor my bones. Can you explain? PMID- 19554748 TI - [Epidemiologic evaluation of malaria in endemic areas]. AB - For decades malarial control has been implemented to control the impact of the disease on the health of populations living in endemic zones. The use of artemisinine combination therapy, intermittent preventive treatment for children and pregnant women, vector-control methods such as long-lasting insecticide impregnated mosquito nets and indoor remanent insecticide spraying has proven to be effective. These practices have lead to such an extensive reduction of the malaria burden in some endemic areas that the objective of eradication that was unimaginable a few years ago is now back to the forefront. Regardless of the method chosen, careful evaluation and surveillance of its effectiveness in man is necessary. Achieving epidemiologic impact is the main goal of malaria control methods. The main measures for evaluation involve parasitological and clinical aspects of human malaria. The purpose of this article is to review methods used for epidemiologic evaluation of malaria burden. PMID- 19554749 TI - [Guidelines for phase III evaluation of vector control methods against malaria]. AB - Most new vector control methods against malaria involve the use of pesticides. Prior to release of these products for general use, their efficacy, persistence, and cross-resistance must be tested on mosquito colonies raised in the laboratory (phase I) then on wild mosquitoes in the field (small-scale), individual dwellings, or experimental huts (phase II). The goal of phase III studies is to evaluate the efficacy and effectiveness of the vector-control product or method against malaria in a population at regular risk for transmission. The main objective of phase III tests is to measure the epidemiologic impact, e.g. on the incidence or prevalence of malaria in humans. This article presents guidelines for carrying out phase III tests of vector-control methods against malaria (e.g. home insecticide spraying or insecticide-impregnated bednet use). It was written by participants in a workgroup formed to define recommendations for the WHOPES (WHO Pesticide Evaluation Scheme). PMID- 19554751 TI - Cytogenetics of the Hessian fly: I. mitotic karyotype analysis and polytene chromosome correlations. AB - The somatic haploid chromosome set of the Hessian fly, Mayetiola destructor (Say),consists of two autosomes and two sex chromosomes. Female somatic cells have pairs of each of these chromosomes (2n = 8). Male somatic cells have two pairs of autosomes and two monosomic sex chromosomes. C-banding revealed pericentromeric heterochromatin on one arm of each chromosome. Mitotic chromosomes were karyotyped using relative lengths, centromeric positions, presence of secondary constrictions, and position and length of C-positive regions. Silver nitrate stained pericentromeric, heterochromatic regions on all four chromosomes but did not stain the entire C-positive regions. In female larvae, four distinct polytene chromosomes were found in the basal region cells of the salivary glands. In male larvae, two distinct and two diffuse polytene chromosomes were found in basal region cells. Differential polytene chromosome morphology in male and female larvae, polytene chromosome indexes, and the position of the nucleolus were used to establish correlations between mitotic and polytene chromosomes. The diffuse morphology of the polytene sex chromosomes in male larvae was probably related to dosage compensation. The number of sex chromosomes in male and female larvae is discussed in relation to sex determination and the production of monogenous progenies. PMID- 19554750 TI - [Methods for the phase IV evaluation of malaria vector control interventions: a case-control study of the effectiveness of long lasting impregnated bed nets after their deployment in Benin]. AB - Vector-control measures are a component of integrated malaria control strategies. After evaluation in phase III pilot studies, these measures are currently being deployed in many endemic malaria zones. Their effectiveness must be evaluated under actual conditions of use but it is not ethically acceptable to use unexposed individuals for control groups. In a attempt to overcome this problem, a case-control study was undertaken to evaluate the effectiveness of long-lasting insecticide treated mosquito nets (LLITN) against clinical malaria attacks due to Plasmodium falciparum in an endemic area of southern Benin. During a 4-month period (July to October 2008), 35 clinically documented cases of uncomplicated malaria (fever + parasite density > 3000/microL) were diagnosed in children less than 5 years old from 6 villages in the Tori Bossito medical district. The parents of these children were interviewed at the same time as the parents of 181 children randomly selected from the same 6 villages. A total of 115 of the randomly selected children who had not been feverish during study period were used as controls. The proportion of children having consistently slept under LLITN throughout the study period was 46% in the case group and 78% in the control group (OR=0.32, 95%CI: 0.15-0.71). These data show that the LLITN provided a significant level of protection, i.e., 68% (IC95%: 29%-85%). This case control study shows that vector control measures can be effectively evaluated after deployment in population. The limitations of this methodology are discussed. PMID- 19554752 TI - Hong Kong research dissemination reports. PMID- 19554753 TI - Optometric storytelling. PMID- 19554754 TI - In-office diagnostic equipment rental. PMID- 19554755 TI - Campus practice management clubs. PMID- 19554756 TI - Infestation by Pyemotes tritici (Acari, Pyemotidae) causes death of stingless bee colonies (Hymenoptera: Meliponina). AB - We report the infestation of stingless bee nests by the mite Pyemotes tritici, which killed four colonies of Tetragonisca angustula and one colony of Frieseomelitta varia in Brazil. The first infected colony, a colony of T. angustula, came from an area between Uberlandia and Araguari, Minas Gerais. The transfer of the mites to the other colonies occurred through the transfer of infected combs and subsequent manipulations. Other colonies in the same meliponary, which had not been manipulated, were not infected. The infestation was terminated by isolating the dead colonies from the meliponary. PMID- 19554757 TI - Cross-training also works in a practice. PMID- 19554758 TI - Media influence on eyewear purchases. PMID- 19554759 TI - New restrictions on Medicare retrospective billing. PMID- 19554760 TI - Effects of stingless bee and honey bee propolis on four species of bacteria. AB - We examined the antibacterial activities of several types of propolis, including Africanized honey bee green propolis and propolis produced by meliponini bees. The antibacterial activity of green propolis against Micrococcus luteus and Staphylococcus aureus was superior to that of Melipona quadrifasciata and Scaptotrigona sp propolis. Only two samples of propolis (green propolis and Scaptotrigona sp propolis) were efficient against Escherichia coli. Melipona quadrifasciata propolis was better than green propolis and Scaptotrigona sp propolis against Pseudomonas aeruginosa. We concluded that these resins have potential for human and veterinary medicine. PMID- 19554761 TI - Taking drug administration seriously. PMID- 19554762 TI - Genetic variability in Melipona quinquefasciata (Hymenoptera, Apidae, Meliponini) from northeastern Brazil determined using the first internal transcribed spacer (ITS1). AB - Melipona quinquefasciata is a ground-nesting South American stingless bee whose geographic distribution was believed to comprise only the central and southern states of Brazil. We obtained partial sequences (about 500-570 bp) of first internal transcribed spacer (ITS1) nuclear ribosomal DNA from Melipona specimens putatively identified as M. quinquefasciata collected from different localities in northeastern Brazil. To confirm the taxonomic identity of the northeastern samples, specimens from the state of Goias (Central region of Brazil) were included for comparison. All sequences were deposited in GenBank (accession numbers EU073751-EU073759). The mean nucleotide divergence (excluding sites with insertions/deletions) in the ITS1 sequences was only 1.4%, ranging from 0 to 4.1%. When the sites with insertions/deletions were also taken into account, sequence divergences varied from 0 to 5.3%. In all pairwise comparisons, the ITS1 sequence from the specimens collected in Goias was most divergent compared to the ITS1 sequences of the bees from the other locations. However, neighbor-joining phylogenetic analysis showed that all ITS1 sequences from northeastern specimens along with the sample of Goias were resolved in a single clade with a bootstrap support of 100%. The ITS1 sequencing data thus support the occurrence of M. quinquefasciata in northeast Brazil. PMID- 19554764 TI - Sequential hygienic behavior in Carniolan honey bees (Apis mellifera carnica). AB - We examined the sequence, order or steps of hygienic behavior (HB) from pin killed pupae until the removal of them by the bees. We conducted our study with four colonies of Apis mellifera carnica in Germany and made four repetitions. The pin-killing method was used for evaluation of the HB of bees. The data were collected every 2 h after perforation, totaling 13 observations. Additionally, for one hygienic colony and another non-hygienic colony, individual analyses of each dead pupa were made at every observation, including all details, steps or sequences of HB. The bees recognize the cells containing dead pupae within 2 h after perforation, initially making a hole in the capping, which is the beginning of HB. Uncapping of the dead brood cell reached maximum values from 4 to 6 h after perforation; after 24 h, practically all cells were already uncapped. Another variable, called brood partially removed, was analyzed 4 h after perforation, after the cells had been perforated, which involved uncapping, followed by partial or total removal of the brood. Maximum values of brood partially removed were found 10 h after perforation, though such cells could be found up to 48 h after perforation. The most frequent sequence of events in both colonies was: capped cell --> punctured cell --> brood partially removed --> empty cell. A new model of three pairs of recessive genes (uncapping u1, u2 and remover r) was proposed in order to explain the genetic control of the HB in Apis mellifera. We recommend evaluating HB 24 h after perforation and using a correction factor to compensate for control removal levels. We found a series of details of HB, which allow a study of how various factors may affect the sequence of the activities involved in HB and investigation of the genetics that controls this process. PMID- 19554763 TI - Hygienic behavior of the stingless bee Plebeia remota (Holmberg, 1903) (Apidae, Meliponini). AB - We investigated hygienic behavior in 10 colonies of Plebeia remota, using the pin killed method. After 24 h the bees had removed a mean of 69.6% of the dead brood. After 48 h, the bees had removed a mean of 96.4% of the dead brood. No significant correlation was found between the size of the brood comb and the number of dead pupae removed, and there was no apparent effect of the origin and the condition of the colony on the hygienic behavior of the bees. Plebeia remota has an efficiency of hygienic behavior superior to that of three of the other four stingless bee species studied until now. PMID- 19554765 TI - Pollen foraging in colonies of Melipona bicolor (Apidae, Meliponini): effects of season, colony size and queen number. AB - We evaluated the ratio between the number of pollen foragers and the total number of bees entering colonies of Melipona bicolor, a facultative polygynous species of stingless bees. The variables considered in our analysis were: seasonality, colony size and the number of physogastric queens in each colony. The pollen forager ratios varied significantly between seasons; the ratio was higher in winter than in summer. However, colony size and number of queens per colony had no significant effect. We conclude that seasonal differences in pollen harvest are related to the production of sexuals and to the number of individuals and their body size. PMID- 19554766 TI - Production of workers, queens and males in Plebeia remota colonies (Hymenoptera, Apidae, Meliponini), a stingless bee with reproductive diapause. AB - Queen, male and worker production was studied during one year in three Plebeia remota colonies from Atlantic Rainforest in Cunha, Sao Paulo State, and two from a subtropical Araucaria forest in Prudentopolis, Parana State. All the colonies were kept in Sao Paulo city during our study. Plebeia remota has reproductive diapause during autumn and winter, which makes its biology of special interest. Brood production begins before spring, renewing the colony cycle. We sampled brood combs monthly in these five colonies. The number of cells in each comb varied significantly with time of the year; the smallest brood combs appear to be a consequence of reduced food availability. However, worker, queen and male frequencies did not differ significantly in time, and this presumably is due to the fact that they all are necessary for the growth, maintenance and reproduction of the colony. Although some molecular, morphological and behavioral differences have been detected in several studies comparing populations from Cunha and from Prudentopolis, we did not find significant differences between the colonies from these two localities in number of brood cells and worker, queen and male production. PMID- 19554767 TI - The impact of forest exploitation on Amazonian stingless bees (Apidae, Meliponini). AB - The protocols available to sustainably exploit natural forest resources are known as "sustainable forest management". This type of management generally does not take into account the effect of timber exploitation on pollinators. Stingless bees, which include many species that play an important role as pollinators and are quite diverse in the Amazon, preferentially make their perennial nests in the base of hollow trees. Normally, during sustainable exploitation of trees, hollow trees are not cut down; however, predatory exploitation of such trees could severely affect natural populations of this pollinator group. PMID- 19554768 TI - Unsuccessful attacks dominate a drone-preying wasp's hunting performance near stingless bee nests. AB - Bee males (drones) of stingless bees tend to congregate near entrances of conspecific nests, where they wait for virgin queens that initiate their nuptial flight. We observed that the Neotropical solitary wasp Trachypus boharti (Hymenoptera, Cabronidae) specifically preys on males of the stingless bee Scaptotrigona postica (Hymenoptera, Apidae); these wasps captured up to 50 males per day near the entrance of a single hive. Over 90% of the wasp attacks were unsuccessful; such erroneous attacks often involved conspecific wasps and worker bees. After the capture of non-male prey, wasps almost immediately released these individuals unharmed and continued hunting. A simple behavioral experiment showed that at short distances wasps were not specifically attracted to S. postica males nor were they repelled by workers of the same species. Likely, short-range prey detection near the bees' nest is achieved mainly by vision whereas close-range prey recognition is based principally on chemical and/or mechanical cues. We argue that the dependence on the wasp's visual perception during attack and the crowded and dynamic hunting conditions caused wasps to make many preying attempts that failed. Two wasp-density-related factors, wasp-prey distance and wasp-wasp encounters, may account for the fact that the highest male capture and unsuccessful wasp bee encounter rates occurred at intermediate wasp numbers. PMID- 19554769 TI - Interaction and behavior of virgin and physogastric queens in three Meliponini species (Hymenoptera, Apidae). AB - We studied the behavior of virgin queens of the stingless bee species Schwarziana quadripunctata, Paratrigona lineata and Tetragona clavipes, investigating internal nest activities, including the cell provisioning and oviposition process. We made direct observation of queen behavior, with the aid of video filming. Forty-four virgin queens of S. quadripunctata were observed; one was larger and more attractive than the others. Miniature queens were more abundant than normal-size queens; both were found in prison chambers. Agonistic behavior between virgin and physogastric queens of P. lineata was observed during attempts at queen supersedure. After the disappearance of the physogastric queen and the appearance of a virgin queen in T. clavipes nests, the brood cells were sealed with pollen alone, but no egg. In all three species, the presence of one or more virgin queens appeared to make the colonies nervous, even though constant production of virgin queens is vital to the survival of the colony and is part of the colony cycle in these bees. PMID- 19554770 TI - Morphometric and genetic changes in a population of Apis mellifera after 34 years of Africanization. AB - Though the replacement of European bees by Africanized honey bees in tropical America has attracted considerable attention, little is known about the temporal changes in morphological and genetic characteristics in these bee populations. We examined the changes in the morphometric and genetic profiles of an Africanized honey bee population collected near where the original African swarms escaped, after 34 years of Africanization. Workers from colonies sampled in 1968 and in 2002 were morphometrically analyzed using relative warps analysis and an Automatic Bee Identification System (ABIS). All the colonies had their mitochondrial DNA identified. The subspecies that mixed to form the Africanized honey bees were used as a comparison for the morphometric analysis. The two morphometric approaches showed great similarity of Africanized bees with the African subspecies, Apis mellifera scutellata, corroborating with other markers. We also found the population of 1968 to have the pattern of wing venation to be more similar to A. m. scutellata than the current population. The mitochondrial DNA of European origin, which was very common in the 1968 population, was not found in the current population, indicating selective pressure replacing the European with the African genome in this tropical region. Both morphometric methodologies were very effective in discriminating the A. mellifera groups; the non-linear analysis of ABIS was the most successful in identifying the bees, with more than 94% correct classifications. PMID- 19554772 TI - Intrinsic colony conditions affect the provisioning and oviposition process in the stingless bee Melipona scutellaris. AB - The cell provisioning and oviposition process (POP) is a unique characteristic of stingless bees (Meliponini), in which coordinated interactions between workers and queen regulate the filling of brood cells with larval resources and subsequent egg laying. Environmental conditions seem to regulate reproduction in stingless bees; however, little is known about how the amount of food affects quantitative sequences of the process. We examined intrinsic variables by comparing three colonies in distinct conditions (strong, intermediate and weak state). We predicted that some of these variables are correlated with temporal events of POP in Melipona scutellaris colonies. The results demonstrated that the strong colony had shorter periods of POP. PMID- 19554773 TI - Climatic and anthropic influence on size and fluctuating asymmetry of Euglossine bees (Hymenoptera, Apidae) in a semideciduous seasonal forest reserve. AB - We examined the influence of climate and man on size and fluctuating asymmetry in two species of Euglossine bees collected from a semideciduous forest reserve. Sixty males of each species were collected; four measurements were made of their wings to obtain a multivariable size index and a fluctuating asymmetry index. No significant differences in the size of Eulaema nigrita Lepeletier were found between the areas and seasons. Larger males of Euglossa pleosticta Dressler were collected during the hot and wet season; however, male size did not vary with location. Higher rainfall and a consequent increase in food availability could have influenced the increase in size of E. pleosticta. Bees collected during the hot and wet season at the forest border were more asymmetric than bees collected during the cold and dry season; the latter were found inside the forest. This indicates that climate and anthropic interferences influence the stability of development of E. pleosticta. Consequently, this species could be used as a bioindicator of stress. Apparently, E. nigrita is more resistant to environmental interference. PMID- 19554774 TI - The first record on Brazilian stingless bees published 450 years ago by Hans Staden. AB - Only a few decades after 1492, when Christopher Columbus arrived on a Caribbean island and Pedro Alvares Cabral claimed Brazil for Portugal in 1500, a German mercenary gave the first description of stingless bees in 1557. He got to know them when he was imprisoned for months by an anthropophagous tribe in the coastal region of Santos, today in the State of Sao Paulo. This rather short but nevertheless extremely exact record on stingless bees is hidden in the first book on Brazil. Three species and important aspects of their life history were treated. This early description has been completely overlooked by bee scientists until now. My note intends to close this evident gap. PMID- 19554771 TI - Africanized honey bees are efficient at detecting, uncapping and removing dead brood. AB - The hygienic behavior of honey bees is based on a two-step process, including uncapping and removing diseased, dead, damaged, or parasitized brood inside the cell. We evaluated during periods of 1 h the time that hygienic and non-hygienic colonies of Africanized honey bees spend to detect, uncap and remove pin-killed brood using comb inserts with transparent walls placed in observation hives. We observed that hygienic colonies are significantly faster in detecting, uncapping and removing dead brood in the cells (P < 0.001). PMID- 19554775 TI - Nursing shortages around the world, HIV/AIDS in Africa, and breastfeeding behaviors. PMID- 19554776 TI - [Intraureteral foreign body]. AB - OBJECTIVE: We present a rare localization for a foreing body in the urinary system, and review the bibliography to know about it existence, frequency and localization. METHODS: We present the case of a 49 year old woman who referreds she had introduced a metallic objet in her genital area. RESULTS: After the necessary tests, we could see a metallic cylinder. It was 8 cm long and was localized inside the right pelvic ureter. CONCLUSIONS: The presence of strange bodies in the urinary system is a rare urological emergency. Treatment is the key element of the extraction of the least bloody way possible. PMID- 19554777 TI - [The genetic variability and clinical significance of hepatitis B virus]. PMID- 19554778 TI - [Bringing the antiviral therapy home for chronic hepatitis B patients: host immunity influences the efficacy and disease prognosis]. PMID- 19554779 TI - [Segmentary testicular infarction]. AB - OBJECTIVE: A new case of segmental testicular infarction is reported. METHODS: A 52-year-old man presented to the emergency department with severe pain in the left testicle over a few days period. Physical examination revealed a tender induration in the upper pole of the left testicle. Beta-human chorionic gonadotropin and alpha-fetoprotein were normal. Sonography demonstrated a poorly defined, hypoechoic, 13 mm lesion. On power Doppler sonography, the hypoechoic area appeared completely avascular in contrast to the rest of the testicle. RESULTS: Since testicular tumor was the initial diagnosis, the patient underwent a left radical orchiectomy. Pathological study revealed a focal testicular infarction, without signs of malignancy or vasculitis. CONCLUSIONS: Segmental testicular infarction is usually diagnosed after radical orchiectomy, performed when testicular tumor is suspected. On certain occasions Doppler ultrasound and magnetic resonance imaging findings suggested a segmental testicular infarction. However, if tumor cannot be entirely excluded, exploratory surgery is necessary. PMID- 19554780 TI - [Biochemical terminology]. PMID- 19554781 TI - [Malignant priapism and secondary bladder cancer]. AB - OBJECTIVE: We report a rare case of malignant priapism secondary to transitional cell carcinoma. METHODS: The patient with bladder cancer (pT4G3) presented with painful penile erection. Corpora cavernosa biopsy was done. RESULTS: The pathologic diagnosis was penile metastasis of transitional cell carcinoma. CONCLUSIONS: Priapism secondary to penile metastasis of transitional cell carcinoma is rare and indicates advanced disease with a poor prognosis. PMID- 19554782 TI - [Spermatic cord sarcomas: current status and report of four cases]. AB - OBJECTIVE: To study and review spermatic cord sarcomas, including symptoms, diagnosis, and treatment. METHODS/RESULTS: We review the Spanish and international literature and report 4 new cases: 2 patients with well differentiated spermatic cord liposarcomas (1 treated by simple tumorectomy), 1 patient with liposarcomatous degeneration of a previously excised atypical lipoma, and 1 patient operated for a malignant retroperitoneal fibrous histiocytoma with subsequent local recurrence in the paratesticular region. CONCLUSIONS: Spermatic cord sarcomas are rare entities that usually appear as painless paratesticular mass. They are diagnosed by imaging studies (ultrasound, computed tomography, magnetic resonance) and confirmed by histological examination. Spermatic cord sarcomas are treated surgically; the efficacy of adjuvant treatments such as chemotherapy or radiation therapy is still under debate. PMID- 19554783 TI - [Calyx diverticulum]. PMID- 19554784 TI - [Fibrous pseudotumor of spermatic cord. A rare entity]. PMID- 19554785 TI - [Evidence based medicine in dietary supplements]. PMID- 19554787 TI - Research from the National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases, 2008 STEP UP High School Program with Charles Drew University of Medicine and Science. PMID- 19554786 TI - [Role of hypoxemia inducible transcription factor in tissue remodeling]. PMID- 19554788 TI - [Intracranial tumor associations: coexistence of low-grade glioma, glioblastoma multiforme, and meningioma in the same patient]. PMID- 19554789 TI - Orbital Langerhans cell histiocytosis with focus on treatment. PMID- 19554790 TI - Pediatric surgery. PMID- 19554791 TI - Reproductive endocrinology and infertility. PMID- 19554792 TI - The whole world is watching. PMID- 19554793 TI - The real lessons of stem cells. PMID- 19554794 TI - Interview with Kimberly A. Russel, FACHE, President and Chief Executive Officer, BryanLGH Health System. Interview by Kyle A Grazier. PMID- 19554795 TI - Field-tested strategies for physician recruitment and contracting. PMID- 19554796 TI - Keeping green: reducing energy consumption without harming the budget. PMID- 19554797 TI - Hospital cost and efficiency: do hospital size and ownership type really matter? AB - The primary research question this study addresses is, do size and ownership type make a difference in the efficiency and cost results of hospitals in Washington State? A further question is, what factors might explain such differences? The data source is the hospital financial data reports Washington hospitals submit to the Washington Department of Health. The sample was restricted to not-for-profit and government-owned hospitals, given that these ownership types are predominant in Washington State, and there are only two investor-owned hospitals. The measures of efficiency and cost represent the generally accepted financial indicators derived from the healthcare financial management literature. Cost and efficiency in these hospitals are analyzed using five efficiency ratios and five cost measures. The results are significant for five of the ten measures studied. Measured by occupancy percentage, small and large not-for-profit hospitals appear to achieve higher efficiency levels than government-owned hospitals do, but the larger hospitals of both ownership types report greater efficiency than that achieved by smaller hospitals. In terms of costs, small, not-for-profit hospitals report comparable costs to those of the largest hospitals, likely because 70 percent of the small not-for-profits are critical access hospitals. These findings deserve further study on a regional or national level. A more scientific study of the efficiency and cost of hospitals by size and ownership type would be important to control for case mix, scope of services, and payer mix. Such studies can generate important findings about the relationship of hospital size and ownership type to efficiency and cost. Conducted on a national level, such studies would provide policymakers with the empirical data they need to make decisions regarding the types of hospitals to encourage or discourage in the future. PMID- 19554798 TI - The relationship between patient satisfaction and inpatient admissions across teaching and nonteaching hospitals. AB - The need for healthcare executives to better understand the relationship between patient satisfaction and admission volume takes on greater importance in this age of rising patient expectations and declining reimbursement. Management of patient satisfaction has become a critical element in the day-to-day operations of healthcare organizations pursuing high performance. This study is guided by two principal research questions. First, what is the nature of the relationship between patient satisfaction (as measured by scored instruments) and inpatient admissions in acute care hospitals? Second, does the relationship between patient satisfaction (as measured by scored instruments) and inpatient admissions differ between teaching hospitals and nonteaching hospitals? Although not suggestive of direct causation, the study findings revealed a statistically significant and positive correlation between patient satisfaction and admission volume in teaching hospitals only. In contrast, a nonsignificant, negative correlation was seen between patient satisfaction and admission in nonteaching hospitals. In the combined teaching and nonteaching sample, a statistically significant, negative correlation was found between patient satisfaction scores and admission volume. With financial performance being driven in part by admission volume and with patient satisfaction affecting hospital patronage, the business case for a strategic focus on patient satisfaction in teaching hospitals is clearly evident. The article concludes with a set of recommendations for strengthening patient satisfaction and organizational performance. PMID- 19554799 TI - Strategic, political, and cultural aspects of IT implementation: improving the efficacy of an IT system in a large hospital. AB - Healthcare spending will exceed $4 trillion by 2017, a trend that is leading executives to implement information technology (IT) systems to contain these rising costs. Studies show that numerous factors determine the outcome and net benefits of IT in healthcare. However, what happens when a newly implemented IT system results in negative outcomes? We explore this question by examining a newly implemented IT system in a large hospital that was yielding none of the benefits for which its designers had hoped. Using an expanded set of analytic lenses, our in-depth study found that political issues were a major stumbling block to the implementation of this IT system, as the interests of IT managers were different from those of the system's users. In addition, cultural values among these stakeholders were not aligned. The new IT system carried very different meanings for these two key groups. These political and cultural issues, which reflect a broader set of factors than is commonly applied in IT or in management, led to specific recommendations designed to improve the system's viability and benefits. In a follow-up analysis we found that these alternative lenses helped increase the intended usage of the IT system by 16 percent in the first year, yielding a 20 percent improvement in performance. By better understanding the cultural and political significance of IT implementation, managers may thus improve the effectiveness of new information technologies for containing costs in hospitals. PMID- 19554800 TI - Unbalanced nutrition might be a threat to our endocrine system. PMID- 19554801 TI - Graves' disease--acceleration of linear growth. PMID- 19554802 TI - Assessing inter-rater reliability (IRR) of Tanner staging and orchidometer use with boys: a study from PROS. AB - BACKGROUND: Few studies have systematically assessed the reliability of pubertal markers; most are flawed by limited numbers of markers and ages studied. AIM: To conduct a comprehensive examination of inter-rater reliability in the assessment of boys' sexual maturity. SUBJECTS: Eight pairs of practitioners independently rated 79 consecutive boys aged 8-14 years. METHODS: Two raters in each of eight practices independently rated boys aged 8-14 years, presenting for physical examinations, on key pubertal markers: pubic hair and genitalia (both on 5-point Tanner scales), testicular size (via palpation and comparison with a four-bead Prader orchidometer), and axillary hair (via a three-point scale). RESULTS: Intraclass correlations assessing degree of inter-rater reliability for pubertal markers ranged from 0.61 to 0.94 (all significant at p < 0.001). Rater Kappas for signs of pubertal initiation ranged from 0.49 to 0.79. CONCLUSIONS: Practitioners are able to reliably stage key markers of male puberty and identify signs of pubertal initiation. PMID- 19554804 TI - Normal adult height among girls treated for central precocious puberty with gonadotropin-releasing hormone analog therapy. AB - AIM: To evaluate adult height (AH) among girls with central precocious puberty (CPP) treated with gonadotropin releasing hormone analog (GnRHa) and to assess the impact of posttreatment growth on AH. STUDY DESIGN: Medical records of girls with CPP were reviewed. RESULTS: Twenty-six girls aged 7.2 +/- 2.0 years were identified. There was a significant difference between AH and predicted adult height (PAH) at the initiation of therapy (p = 0.005). Using univariate analysis, the only factor associated with AH was total growth after discontinuation of therapy. Growth after discontinuation of therapy was variable and often greater than expected. Both age and skeletal age at the end of therapy had strong linear relationships with growth after therapy explaining 60% of this growth. CONCLUSION: This report confirms that AH is normal among females with CPP treated in a timely fashion with GnRHa. The lack of predictability of growth after discontinuation of therapy suggests that the decision to stop treatment should be individualized. PMID- 19554803 TI - Body composition in premature adrenarche by structural MRI, 1H MRS and DXA. AB - BACKGROUND: Premature adrenarche (PA) is recognized to be a possible precursor of polycystic ovarian syndrome, type 2 diabetes mellitus and cardiovascular disease. Visceral adiposity and increased intramyocellular lipid (IMCL) are associated with insulin resistance and increased risk of cardiovascular disease. AIM: To determine whether prepubertal girls with PA have altered visceral adiposity and/or increased muscle lipid content compared to prepubertal girls without PA using proton magnetic resonance imaging (MRI) and spectroscopy (1H MRS). PATIENTS AND METHODS: We performed total body dual energy X-ray absorptiometry (DXA) scans, MRI of the trunk, and MRS of the tibialis anterior muscle in the right calf on six girls with PA and eight prepubertal controls. RESULTS: Amount of visceral adipose tissue (VAT), abdominal subcutaneous adipose tissue (SAT), and VAT to SAT ratio did not differ significantly between the PA and control girls. Those with PA, however, had significantly greater IMCL than controls (p = 0.004). CONCLUSIONS: This study adds further evidence that PA is not a benign condition, and future studies investigating early intervention with dietary and exercise counseling may help diminish potential risk for diabetes mellitus and/or cardiovascular disease. PMID- 19554805 TI - Etiological factors for the persistence of endemic goiter in selected areas of Siddharthnagar district in Eastern Uttar Pradesh, India. AB - AIM: To assess the prevalence of goiter, state of iodine nutrition of the population, consumption pattern of common goitrogenic food, and distribution of iodine through edible salt in selected CD Blocks of Siddharthnagar district in Eastern Uttar Pradesh. METHODS: Goiter survey among 1,862 school children, aged 6 12 years, of both sexes, and analysis of iodine (I) and thiocyanate (SCN) in 240 urine samples, and iodine content in 210 edible salt samples collected from the selected study areas. RESULTS: The prevalence of goiter was 26.3% (grade 1: 23.2%; grade 2: 3.1%). Median urinary iodine level was 6.0 microg/dl, and 42% had concentration < 5 microg/dl. Mean (SD) urinary SCN was 0.75 (0.4) mg/dl. Only17.1% of salt samples had iodine level > 15 ppm; 82.9% had < 15 ppm. CONCLUSION: Consumption of inadequately iodized salt and cyanogenic plant foods containing goitrogenic/anti-thyroidal substances by the people of the studied region are possible reasons for the persistence of goiter during the post salt iodination phase. PMID- 19554806 TI - Excessive iodine intake and ultrasonographic thyroid abnormalities in schoolchildren. AB - High nutritional levels of iodine may induce a higher prevalence of autoimmune thyroiditis, hypothyroidism, goiter, as well as hyperthyroidism, mostly in the elderly. This study assessed thyroid volume and ultrasonographic abnormalities as well as urinary iodine excretion (UIE) in 964 schoolchildren living in an iodine sufficient area in southern Brazil. Thyroid volume correlated with age and body surface area in boys and girls. In 76.8% of the children, UIE was above 300 microg/l, with higher levels among boys compared to girls (484.2 microg/l vs 435.3 microg/l, p < 0.001). Thyroid abnormalities detected by ultrasonography included hemiagenesis (0.5%), nodules (0.2%), cysts (0.7%), and hypoechogenicity (11.7%). Goiter was present in 1.9% of the children. Hypoechogenicity, a relevant marker of autoimmune thyroiditis, was the most common abnormality found in our study, and this may be linked to excessive iodine intake. PMID- 19554807 TI - Higher frequency of screening TSH above 5 mIU/l in infants likely exposed to higher doses of iodine-containing skin antiseptic: implications for assessment of iodine sufficiency. AB - BACKGROUND: Neonatal TSH screening has as its main goal the early detection and treatment of permanent sporadic congenital hypothyroidism. At the same time neonatal TSH is one of the indicators for monitoring progress towards eliminating iodine deficiency disorders (IDD). AIM: To evaluate the usefulness of neonatal TSH screening data as a monitoring tool for IDD evaluation and control in Bosnia and Herzegovina. INFANTS AND METHODS: The study included a total of 8,105 newborns. Neonatal TSH was measured in whole blood drawn between day 3 and day 5 of life, spotted on filter paper using a sensitive fluorometric assay (Delfia). RESULTS: Levels above 5 mIU/l were found in 282 (5.5%) neonates. Median TSH values and the percentage of samples of neonatal TSH above 5 mIU/l were significantly higher in neonates who were born by Caesarean section (6.4%) compared with neonates born by vaginal delivery (1.4%). CONCLUSION: Our findings suggest that neonatal TSH should not be used as a monitoring tool for IDD evaluation and control if iodinated skin disinfectant is used on a large part of the mother's skin in maternity hospitals. PMID- 19554808 TI - Evolution of sonographic appearance of the thyroid gland in children with Hashimoto's thyroiditis. AB - BACKGROUND: Although thyroid ultrasound is a valuable tool for the diagnosis and follow-up of patients with Hashimoto's thyroiditis (HT), classical sonographic findings are not always present. AIM: To calculate the time needed for children with HT and normal ultrasound at diagnosis to develop characteristic sonographic findings. PATIENTS AND METHODS: 105 children (23 male and 82 female) with HT (mean age 9.4 +/- 2.9 years) were studied. Physical examination and measurements of TSH and fT4 levels were performed at diagnosis, at 3-month intervals for the first year, and twice yearly thereafter. Thyroid ultrasound was performed at diagnosis and twice yearly thereafter. The median follow-up duration was 18 months (range: 6-61 months). RESULTS: The time needed for 30%, 50%, and 70% of children to demonstrate an abnormal thyroid sonographic pattern was 4, 7, and 14 months, respectively. Important factors accelerating sonographic changes were goiter (p = 0.023), hypothyroidism (p = 0.0255), and seropositivity for both thyroid peroxidase (anti-TPO) and thyroglobulin (anti-Tg) autoantibodies (p = 0.0005). CONCLUSION: Sonographic findings of HT are present in 37% of children at diagnosis. Fifty percent of children with normal initial thyroid US will develop changes within 7 months; however, characteristic findings may not develop for over 4 years. PMID- 19554809 TI - Metabolic abnormalities in human immunodeficiency virus-infected children: two year follow-up. AB - BACKGROUND: HIV-associated lipodystrophy (LD) manifests with fat maldistribution, dyslipidemia, and insulin resistance in some HIV-infected children on antiretroviral therapy. AIM: To assess whether lipid abnormalities in patients with HIV are stable over time. PATIENTS: The perinatally HIV-infected cohort at a medium-sized urban US teaching hospital. METHODS: This prospective, observational study consisted of five visits (at entry and 3, 6, 24, and 30 months after entry) during which fasting venous blood samples were drawn for HIV-1 RNA, CD4 lymphocytes, lipid profile, free fatty acids (FFA), glucose, insulin, and adiponectin. IGF-I/IGFBP-3 levels were measured at the first and fifth visits. RESULTS: Of 36 study participants, seven were lipodystrophic, and 30 patients completed all five study visits. LDL-cholesterol, total cholesterol (TC), triglycerides (TG), and FFA levels were significantly higher in patients taking protease inhibitors (PIs). Patients with LD had higher TC and TG levels (both p < 0.05), and higher FFA (p = 0.0532). Adiponectin levels did not differ between PI/non-PI and LD/non-LD groups. HDL-cholesterol seemed to decrease, and FFA to increase over time. All IGF-I and all but one IGFBP-3 level were within normal range for age and Tanner stage. CONCLUSION: Dyslipidemia remained relatively constant over our study period. Adiponectin was not useful as a marker of LD in our population. PMID- 19554810 TI - Can the insulin sensitivity index (ISI) in association with insulin-like growth factor binding protein-1 identify insulin resistance early in overweight children? AB - Association between insulin resistance (IR) and non-alcoholic fatty liver disease (NAFLD) has been reported. This prompted us to evaluate the power of the insulin sensitivity index (ISI) in association with IGFBP-1 to identify IR early in obese children/adolescents. OGTT was performed in 34 obese/overweight children/adolescents. Glucose, insulin and IGFBP-1 were measured in serum samples and ISI was calculated. Considering the presence of three or more risk factors for IR as a criterion for IR, ISI < 4.6 showed 87.5% sensitivity and 94.5% specificity in diagnosing IR. IGFBP-1 was lower in the group with ISI < 4.6 (p < 0.01). In this group, three patients had higher than expected IGFBP-1, suggesting hepatic IR, while three patients with ISI > 4.6 showed very low IGFBP-1 levels. CONCLUSION: ISI < 4.6 is a good indicator of early peripheral IR and, associated with IGFBP-1, can identify increased risk of hepatic IR. Low IGFBP-1 levels among non-IR children may indicate increased portal insulin levels. PMID- 19554812 TI - Autoimmune type I diabetes mellitus in a perinatally HIV infected patient with a well-preserved immune system. AB - We report an 8 year-old girl with well-controlled perinatally acquired HIV infection who developed autoimmune type 1 diabetes mellitus (DM1A) confirmed by the presence of diabetes-related auto-antibodies. Although non-autoimmune insulin dependent diabetes mellitus (DM1B) and more frequently type 2 DM has been reported in patients affected with HIV, this is the first report of DM1A diagnosed in an HIV positive patient. PMID- 19554811 TI - Cushing's syndrome due to a non-adrenal ectopic adrenocorticotropin-secreting Ewing's sarcoma in a child. AB - Ectopic ACTH syndrome (EAS) is extremely rare in the pediatric age group. Sarcomatous tumors causing EAS are even rarer. We report a 9 year-old girl presenting with Cushing's syndrome caused by ectopic ACTH production by a Ewing's sarcoma. This case illustrates that rapid appearance of Cushingoid symptoms, absence of growth retardation and the presence of hypokalemia are suggestive clues for ectopic ACTH production as the source of Cushing's syndrome. PMID- 19554813 TI - Evolving diagnosis of McCune-Albright syndrome. atypical presentation and follow up. AB - The diagnosis of McCune-Albright syndrome has classically consisted of the triad of gonadotropin-independent precocious puberty (GIPP), cafe-au-lait (CAL) spots and fibrous dysplasia of bone. Atypical or incomplete forms of the syndrome have been reported in the literature, with only one or two of the classical symptoms present, and the diagnosis being made after molecular analysis. Therefore not all three classical findings need to be present for the diagnosis to be made. We report a patient who was suspected of having McCune-Albright syndrome after presenting initially with GIPP at age 2 years 10 months. At age 10 years 1 month, a CAL spot appeared and, after reviewing previous imaging studies, a bone cyst was found. Molecular analysis in peripheral leucocytes was negative. We reviewed similar cases described in the literature. PMID- 19554814 TI - Acute suppurative thyroiditis as the initial presentation of juvenile systemic lupus erythematosus. AB - We report an unusual case of acute suppurative thyroiditis associated with juvenile systemic lupus erythematosus (JSLE) onset. A 10 year-old girl presented with a painful voluminous swelling in the anterior cervical region with fever and dysphagia. She also had dysphonia, myalgia, swelling of the small joints of the hands, weight loss, and erythematous malar rash. Thyroid ultrasonography disclosed an ill-defined solid cystic area containing abundant echoes in suspension suggestive of an abscess in both lobes compatible with suppurative bacterial thyroiditis. Fine-needle aspiration biopsy showed a secretion with positive culture for Staphylococcus aureus. Endoscopic evaluation did not demonstrate pyriform sinus fistula. The child had no previous thyroid disease. Hormonal evaluation showed thyrotrophin (TSH) 13 microIU/ml and free thyroxine (FT4) 0.4 ng/dl. After laboratory confirmation of JSLE, high dose 'pulse' therapy was performed with methylprednisolone (30 mg/kg/day for 3 days), followed by prednisone (2 mg/kg/day). The acute thyroiditis was treated with oxacillin, metronidazole and amikacin which ameliorated the symptoms of infection followed by normalization of thyroid function with levothyroxine. In spite of the known association between autoimmune diseases and thyroid disorders, the coexistence of suppurative thyroiditis and JSLE is rare, generally occurring during the course of lupus and not at diagnosis. PMID- 19554815 TI - Paradigms for choosing paradigms. PMID- 19554816 TI - The 10-year experience of Oregon's Death with Dignity Act: 1998-2007. PMID- 19554817 TI - Putting Oregon's Death with Dignity Act in perspective: characteristics of decedents who did not participate. PMID- 19554818 TI - Must physicians report impaired driving? Rethinking a duty on a collision course with itself. PMID- 19554819 TI - Is there coercion or undue inducement to participate in health research in developing countries? An example from Rakai, Uganda. PMID- 19554820 TI - Patients' perceptions on healthcare decision making in rural India: a qualitative study and ethical analysis. PMID- 19554821 TI - Patients' concerns for family burden: a nonconforming preference in standards for surrogate decision making. PMID- 19554822 TI - Berger on burdens. PMID- 19554823 TI - Commentary on "Patients' concerns for family burden". PMID- 19554824 TI - Commentary on Berger's "Patients' concerns for family burden". PMID- 19554825 TI - When I lay my burden down: commentary on Berger. PMID- 19554826 TI - What really is in a child' s best interest? Toward a more precise picture of the interests of children. PMID- 19554827 TI - Accommodating religious and moral objections to neurological death. PMID- 19554828 TI - Religious belief and surrogate medical decision making. PMID- 19554829 TI - [Intervention in acute coronary syndrome: not everything at once ]. PMID- 19554830 TI - [Atrial fibrillation]. AB - Atrial fibrillation is the arrhythmia that most frequently leads to hospital admission. As prevalence of atrial fibrillation increases with age, its epidemiological relevance will increase due to the well-known changes in life expectancy. In the presence of atrial fibrillation the cardiovascular mortality and the risk for a stroke are considerably elevated. Interventional treatment, such as catheter ablation or special pacemaker algorithms, have been improved extensively in the last years as a therapeutic option. Nevertheless drug therapy is still the first choice of treating atrial fibrillation. PMID- 19554831 TI - [The Marfan syndrome and related connective tissue disorders]. AB - The Marfan syndrome is an inherited disorder of the connective tissue which is mainly caused by a mutation in the fibrillin-1 gene. The defect in the connective tissue protein can lead to several organ dysfunctions. For the life expectancy, the cardiovascular aspect is of paramount importance. Patients with Marfan syndrome may develop aortic aneurysms and valvular heart defects. The risk of aortic aneurysms consists in the development of aortic dissection or rupture with their fatal consequences. Besides the cardiovascular manifestation, the skeletal and ocular system can also be affected. The skeletal manifestation is often characterised by long limbs, arachnodactyly, and abnormal joint flexibility along with other signs. Patients may also have dislocated lenses, ectasia of the dural sac, stretch marks, spontaneous pneumothorax, recurrent hernia, or a family history suspicious for Marfan. During the past years, other related connective tissue disorders with analogous organ manifestation have been described (e.g., Loeys-Dietz syndrome). In this article we present the basic knowledge about these connective tissue disorders, and we mention new insights in the recently explored pathophysiology of the disorder which is a possible target for future medical treatment options. Furthermore, recent new concepts for the prophylactic treatment of the aortic manifestation are explained. PMID- 19554832 TI - [Presbyacusis: pathogenesis and treatment]. AB - Although many adults retain good hearing as they age, hearing loss associated with ageing is common among elderly persons. There are a number of pathophysiological processes underlying age-related changes to functional components. Presbyacusis is especially caused by cochlear degeneration, most pronounced in the basal cochlear coil. Factors include physiological ageing processes as well as endogenous or exogenous causes. The common form of hardness of hearing seen in old age is not due to physiological age-related changes, but rather to a complex sensorineural pattern of injury. In the industrial countries, two main exogenous causes are exposure to loud noise and obesity. Pathomechanisms contributing to presbyacusis are hypoxia/ischemia, reactive species formation and oxidative stress, apoptotic and necrotic death of hair cells and spiral ganglion cells as well as inherited and acquired mutations in the mitochondrial DNA. Important for the successful treatment of presbyacusis is a timely fitting of hearing aids on both ears to improve communication, provide the auditory system with acoustic information, and potential prevention of social isolation. At present, several therapeutic interventions are under discussion. The application of antioxidants or caloric restriction is considered to prevent or reduce oxidative stress-induced damage. A further approach may be the overexpression or modulation of the superoxide dismutase 2 (SOD2) within the cochlea. Hair cell regeneration could also be a possible treatment of presbyacusis in the future. PMID- 19554833 TI - [Fluconazole for recurrent vulvovaginal candidiasis]. PMID- 19554834 TI - Maine revamps funding formula. PMID- 19554835 TI - More than an ounce of prevention. There's value, wisdom in the push for wellness, but commitment must be sustained. PMID- 19554836 TI - New tourist attractions. More patients looking at home before going abroad. PMID- 19554837 TI - Largest healthcare financing companies. Ranked by amount of syndicated loans in the healthcare sector underwritten in 2008. PMID- 19554838 TI - The sins of the fathers. PMID- 19554839 TI - Five of a kind. PMID- 19554840 TI - From bench to bedside. PMID- 19554841 TI - Skin cancer awareness: not just in May. PMID- 19554842 TI - Promoting prevention and early recognition of malignant melanoma. AB - Research continues to support several of the traditional risk factors for melanoma: a personal history of melanoma, basal or squamous cell carcinomas, the presence of moles, sun sensitivity, occupational exposure to certain substances, or a depressed immune system (ACS, 2007; Batailee et al., 2007; Batistatou et al., 2007). These are now joined by several new risk factors, namely, a history of dysplastic nevi, nevi persisting into adulthood, the use of pesticides, and not being vaccinated with BCG or vaccinia vaccines (Fortes et al., 2007; Krone et al., 2005; Shors et al., 2006). In addition, current pathology and pharmacology studies point towards an endogenous origin for malignant melanomas. Malignant melanoma's profile is expanding on a genotypic and phenotypic level. New evidence shows increasing rates of melanoma in minority ethnicities, especially Asians and Hispanics, people in lower socioeconomic groups, as well as elderly White men (Hu et al., 2006; Reyes-Ortiz et al., 2006); therefore, health care practitioners should screen these high-risk demographic groups more closely (Cormier et al., 2006; Hu et al., 2006; Reyes-Ortiz et al., 2006). Also, more educational materials tailored to these at-risk populations, especially minorities and the elderly, need to be formulated as the majority of melanoma awareness materials are created to target the middle-aged Caucasian demographic (Cockburn et al., 2006; Hu et al., 2006). Most PCPs are not performing skin exams regularly, and lack of time and confidence are major reasons for this omission (Geller et al., 2004). This finding underscores Pender's theory that if practitioners have a perceived higher confidence in their ability to perform cutaneous exams, then they are more likely to perform routine full skin exams and sun-protection education (Pender et al., 2002). Pender makes it clear that patients will act in their own best interest, changing dangerous behaviors and increasing healthy practices if they have the knowledge, tools, and incentives to do so (Pender et al., 2002). Thus, practitioners need to educate patients on the benefits of sun avoidance and regular skin exams. In this way, providers function as a vital influence on a patient's own adaptation of health-promoting behaviors (Pender et al., 2002). Primary care providers need to help remove the barriers of social prejudice through campaigning for stricter legislation, which will tighten regulations on indoor tanning salons. Whitehead (2004) notes, "More emphasis on policy-driven initiatives that work through social examination and modification" lead to a community health-promoting empowerment (p. 314). According to Pender and colleagues (2002), communal health awareness is necessary for an individual to make a solid commitment to a health-promoting behavior. Hopefully, as practitioners become more aware of the major risk factors for melanoma, the rate of melanoma prevention for at-risk patients will increase. This review has served to highlight those risk factors, and the tools which practitioners have to decrease melanoma's morbidity and mortality: appropriately identifying high-risk patients, performing in-office annual skin exams, offering patient education regarding self-skin exams and sun-protective behaviors, and pushing for stronger restrictions on the indoor tanning industry. If practitioners can skillfully utilize these tools, then a significant achievement against the progression of malignant PMID- 19554843 TI - The evidence on how to best treat sunburn in children: a common treatment dilemma. PMID- 19554844 TI - Nursing research: tips and tools to simplify the process. PMID- 19554845 TI - What's your assessment? A permanent, non-tender skin discoloration. Nevus of Ito. PMID- 19554847 TI - For patients, visits aren't routine. PMID- 19554846 TI - Vaseline: a historical perspective. PMID- 19554848 TI - Misadventures in cosmetic dermatology. PMID- 19554850 TI - Hospitality and service: leading real change. PMID- 19554849 TI - Disseminated zoster. PMID- 19554852 TI - Is there really such a thing as hope in a jar? PMID- 19554851 TI - How you feel on the inside could be affecting how you look on the outside. PMID- 19554853 TI - Vexatious litigant law reform. AB - A number of jurisdictions, including in the United Kingdom, Canada, Australia, India and Hong Kong, have recently canvassed reform of vexatious litigant laws which in prescribed circumstances restrict access to the litigation system. The most recent contribution to law reform on the subject is the 2008 report of the Law Reform Committee of the Victorian Parliament. Having reviewed psychiatric and psychological literature on relevant mental health considerations, and taken into account empirical data on the subject, as well as having engaged in extensive consultations, the Committee recommended a series of graduated litigation restriction orders, comparable to but distinct from the system in place in the United Kingdom. This editorial reviews the recommendations of the Committee, the reasons for them and their advantages. It argues that there is much to be said for law reform in relation to vexatious litigation that has, at its centre, management of difficult litigants in a humane and responsive way and that has resort to preclusionary orders only to the minimum extent necessary to guard against foreseeable harm from the repeated bringing of unmeritorious applications or litigation. PMID- 19554854 TI - Prevention of suicide: police powers, parliamentary intent and judicial interpretation. AB - While in most countries suicide is no longer a crime, it is also acknowledged that the state has an interest in the preservation of human life, the prevention of suicide, and the protection of vulnerable persons from harming themselves. In a civil, secular and democratic society, however, the public law principle of state protective powers has to be balanced against the private law principle of personal autonomy (personal self-determination). Under the doctrine of autonomy, competent adults of sound mind can make legally binding voluntary choices, including the so-called "death-choice" (refusal of life-sustaining or life prolonging treatment as well as suicide). To add to the complexity, whereas the powers of the state in relation to suicide and its prevention have been codified, the concepts of personal autonomy and personal liberty are grounded in common law. Kirkland-Veenstra v Stuart [2008] Aust Torts Reports 81-936; [2008] VSCA 32, which is at present being considered by the High Court of Australia, exemplifies tensions that arise in the suicide-prevention area of jurisprudence. This article explores the powers and duties of police officers in relation to suicide prevention and the notion of mental illness by reference to the Kirkland-Veenstra case, the relevant statutory framework and the common law. PMID- 19554855 TI - Support of victims of offenders with mental illness. AB - The rights of victims of crime, particularly the right to be heard and have their experience validated, are often neglected in the criminal justice process. Where the offender is diverted into the mental system for treatment and particularly when the offender subsequently avails of a mental health defence and is acquitted, victims may not understanding the process and may feel aggrieved at the outcome. When an offender with a mental illness responds to treatment and rehabilitation and begins to negotiate leave from hospital preparatory to reintegration into the community, it may be very difficult for victims to accept the transition. As a result of the recommendations of the Butler Review of the Mental Health Act in Queensland, there are now quite innovative provisions which recognise the rights of victims and further enhance the confidence of the public in the safety, transparency and accountability of the process of disposition and community leave of offenders with mental illness. The provisions may provide a model for other jurisdictions. PMID- 19554856 TI - Whose best interests? Advance directives and clinical discretion. AB - A patient's advance directive (AD), confronts a clinician with a quasi-legal document of uncertain status in different jurisdictions and therefore a challenge to her or his clinical acumen and skill. The permutations of factors that need to be taken into account in the resulting clinical decision can be quite confusing. The uncertainties that arise in relation to the provision or otherwise of various forms of invasive and possibly risky clinical treatment can be very disconcerting in that the clinician may be aware of the need to account for the appropriateness of judgments about life-saving treatment and not just the clinical competence that was exhibited. The clinician needs to be clear about the relationships between the duty of care, reasonable medical practice, consent, and what would be considered a substantial benefit or unacceptable risk to the patient. This column outline a decision structure that treats reasonable practice and the patient's wishes as combining to determine whether or not clinical interventions should proceed. PMID- 19554857 TI - The impact of nursing numbers and skills mix on patient outcomes. AB - This column reviews a recent inquest conducted by the Northern Territory coroner into the death of a patient in the Royal Darwin Hospital (Inquest into the Death of Margaret Winter [2008] NTMC 049). It analyses the impact on patient outcomes of the decisions that are made about nursing numbers and the most appropriate skills mix in particular situations. It also looks at the decision-making processes involved in making such assessments. PMID- 19554858 TI - Recent legal developments and the authority of the Australian Therapeutic Goods Administration. AB - Recent legal developments have highlighted the need for greater support from the Federal Government for the authority of the Therapeutic Goods Administration (TGA) to ensure, by pre-approval assessments and post-approval regulation, the safety of listed medicines in Australia. One of these developments concerns the impact of ongoing civil litigation in Australian courts led by Pan Pharmaceuticals stakeholders to recover compensation from the government for the losses they incurred following the TGA's post-listing shut-down of that pharmaceutical manufacturing company in 2003. Another factor is the recently announced governmental policy to outsource to the United States Food and Drug Administration (FDA) and the European Medicines Agency (EMEA) safety assessments of foreign drug manufacturers whose products will be used in Australia. PMID- 19554859 TI - Genetic discrimination in the workplace: towards legal certainty in uncertain times. AB - Scientists can analyse DNA to reveal our likelihood of developing certain diseases. As technology advances, it raises questions as to how much emphasis should be placed on genetic testing in the workplace. In 2008 the United States implemented the Genetic Information Nondiscrimination Act, prohibiting employers from using a person's genetic information in the workplace. This article discusses how Australia should act to prevent unjustified genetic discrimination in the workplace. In 2003 a report from the Australian Law Reform Commission and the Australian Health Ethics Committee, entitled Essentially Yours, concluded it was important to legislate now to protect employees from unlawful discrimination rather than wait for the technology to advance. This article assesses and adds to the recommendations of this report. It argues that genetic status should be considered an inherent requirement of a job, and therefore genetic discrimination should be lawful where it may protect employees themselves, third parties at risk and in professions where it is necessary that employees be healthy in the future. However, to avoid unjustified discrimination, genetic information should only be lawfully used in the workplace where there is a very high probability that the disease will manifest within a specified number of years. PMID- 19554860 TI - Competency and capacity: the legal and medical interface. AB - The loss of legal competency, in the context of wills, enduring powers of attorney and advance directives, presents a challenge to individual autonomy. Both legal and medical practitioners have roles to play in determining when, and if, to infringe upon a person's sovereignty in order to provide for their protection. However, there is some evidence that inter-professional discussions are characterised by tension. Medical expertise is necessary to assess physical and mental capacity, but the legal concern is with competency, two distinct terms. It is argued here that cooperation between the legal and medical professions is essential in this area of practice. This article attempts to promote discussion of this objective by proposing therapeutic jurisprudence as a theoretical framework in which to reassess competency determinations, by reviewing the institutional structure and by suggesting that the adoption of a common and consistent terminology is essential. PMID- 19554861 TI - Emerging legal concerns with chronic diseases in the Australian workplace: pre employment medicals, functional capacity evaluations, workers' compensation and disability discrimination. AB - Although considerable attention is paid to injury and disease sustained or contracted through work causes, by far the most prolific cause of diminished health in the Australian workplace is chronic illness, which is not caused by work but which may have significant effects upon the work environment. Employers' concerns in relation to loss of productivity through poor health are reflected in a number of practices such as pre-employment screening, on-the-job drug testing and other health and safety requirements. In turn, workers' concerns regarding discrimination, workers' compensation and privacy are frequently raised in relation to discussions on workplace fitness for work. This article reflects on the issues of chronic illness and the legal issues which arise through the interaction of employers' obligations for safety and efficiency and workers' concerns with fairness and privacy. PMID- 19554862 TI - Assessing the safety and cost-effectiveness of early nanodrugs. AB - This article provides a detailed examination of how the safety and cost effectiveness elements of Australia's drug regulatory system will respond to nanomedicines. The case study investigated involves Abraxane, a newly developed anti-cancer agent. The article concludes by proposing some responses to the challenges which nanomedicines are likely to present to international and domestic agencies. Additionally, it considers whether the recommendation of the Australian Productivity Commission to allow parallel submissions to the Pharmaceutical Benefits Advisory Committee (PBAC) and the Therapeutic Goods Administration (TGA) is appropriate when applied to new nanotherapeutics. PMID- 19554863 TI - Hindsight bias and outcome bias in the social construction of medical negligence: a review. AB - Medical negligence has been the subject of much public debate in recent decades. Although the steep increase in the frequency and size of claims against doctors at the end of the last century appears to have plateaued, in Australia at least, medical indemnity costs and consequences are still a matter of concern for doctors, medical defence organisations and governments in most developed countries. Imprecision in the legal definition of negligence opens the possibility that judgments of this issue at several levels may be subject to hindsight and outcome bias. Hindsight bias relates to the probability of an adverse event perceived by a retrospective observer ("I would have known it was going to happen"), while outcome bias is a largely subconscious cognitive distortion produced by the observer's knowledge of the adverse outcome. This review examines the relevant legal, medical, psychological and sociological literature on the operation of these pervasive and universal biases in the retrospective evaluation of adverse events. A finding of medical negligence is essentially an after-the-event social construction and is invariably affected by hindsight bias and knowledge of the adverse outcome. Such biases obviously pose a threat to the fairness of judgments. A number of debiasing strategies have been suggested but are relatively ineffective because of the universality and strength of these biases and the inherent difficulty of concealing from expert witnesses knowledge of the outcome. Education about the effect of the biases is therefore important for lawyers, medical expert witnesses and the judiciary. PMID- 19554864 TI - Is there any role for health care professionals at the mediation of clinical negligence claims? AB - It is unusual to see health care professionals present at the mediation of a clinical negligence claim which involves them. Many legal representatives could count on one hand the number of times they have either seen or taken a health care professional to mediation. This is a departure from classical mediation practice, where the parties to the dispute play the primary, if not dominant, role at mediation, including in deciding whether to mediate, making opening statements, identifying issues, developing solutions and deciding whether to resolve the dispute. Although there are a variety of reasons why a health care professional's presence at mediation may not be beneficial, will a health care professional's absence increase the prospects of resolving a dispute? Are there are situations where a health care professional should be present? PMID- 19554865 TI - Assault in medical law: revisiting the boundaries of informed consent to medical treatment in South Africa. AB - This article focuses on assault as a cause of action in medical law, with particular emphasis on the requirements for informed consent, both under common law and under the National Health Act 2003 (Sth Africa). In particular, the test for consent, adopted in Castell v De Greef 1994 (4) SA 408 (C), is analysed in detail. It is noted that the wording of this test for informed consent mirrors the wording of the test for negligence laid down in the Australian case of Rogers v Whitaker (1992) 175 CLR 479. Further, the relationship between the delictual elements of wrongfulness and fault in assault is discussed. It is argued that in South African law a valid consent to medical treatment requires knowledge not only of the general nature of medical treatment but also of the consequences of the treatment and, in determining which consequences should be disclosed to a patient, the constitutional rights to equality and self-determination support the application of a subjective patient-centred test for informed consent. However, it is also proposed that the broad right of a patient to information is reined in at the fault element of assault, so that a limited requirement of consciousness of wrongfulness on the part of the defendant negates liability for delictual assault. PMID- 19554866 TI - New Zealand law's protection of drug trial participants. AB - Pharmaceutical trials are the process by which the effects of new medicines are investigated in order for them to be made safely available to the general public. Because of this unique function, it is important that there be legal protection for those who volunteer to be participants in these trials. This article considers the New Zealand legal framework surrounding pharmaceutical trials, in particular their protection of trial participants. It does this by looking first at the legal regulation of the drug trial process; secondly, at the application of the Code of Health and Disability Services Consumers' Rights; thirdly, by evaluating the adequacy of this legal environment; and finally, by suggesting how it could be improved through law reform. PMID- 19554867 TI - Psychiatric advance directives: the new frontier in mental health law reform in Australia? AB - The legal recognition of psychiatric advance directives is arguably at the forefront of human rights-based mental health law reform in Australia. However, academic discourse in Australia has largely neglected this important development. On the one hand, proponents of psychiatric advance directives believe that such instruments further the rights and autonomy of the mentally ill by allowing consumers of mental health services the right to participate in their own health care when they are competent to make health care decisions. On the other hand, opponents believe that they are undesirable and unworkable in practice and that giving mentally ill persons a right to consent to, or refuse, mental health treatment before the onset of any psychiatric illness does not actually promote or protect the best interests of the mentally ill since future decisions cannot be made about potentially unforeseen circumstances. This article argues that the time has come to consider using psychiatric advance directives in the mental health arena and for amending legislation to be introduced to give them a legal basis. PMID- 19554868 TI - Seven decades of history of science: I. Bernard Cohen (1914-2003), second editor of Isis. AB - I. Bernard Cohen (1914-2003), the first American to receive a Ph.D. in history of science, was a Harvard undergraduate ('37) and then a Ph.D. student and protege of George Sarton, founder of Isis and the History of Science Society. He went on to succeed Sarton as editor of Isis (1952-1958) and, later, president of the Society (1961-1962); he was also a president of the International Union of the History and Philosophy of Science. Cohen was an internationally recognized Newton scholar; his interests were encyclopedic, ranging from science and public policy to the history of computers, with several decades as a special consultant for history of computing with IBM. Among his hundreds of publications were such major books as Franklin and Newton (1956), The Birth of a New Physics (1959; rpt., 1985), The Newtonian Revolution (1980), Revolution in Science (1985), Science and the Founding Fathers (1995), Howard Aiken: Portrait of a Computer Pioneer (1999), and his last book, The Triumph of Numbers (2005), not to mention two jointly authored contributions, the variorum edition and new English translation of Newton's Principia, which will surely still be read a century from now. PMID- 19554869 TI - The practice of Spencerian science: Patrick Geddes's Biosocial Program, 1876 1889. AB - From the Victorian era to our own, critics of Herbert Spencer have portrayed his science-based philosophical system as irrelevant to the concerns of practicing scientists. Yet, as a number of scholars have recently argued, an extraordinary range of reformist and experimental projects across the human and life sciences took their bearings from Spencer's work. This essay examines Spencerian science as practiced by the biologist, sociologist, and town planner Patrick Geddes (1854 1932). Through a close examination of his experimental natural history of the late 1870s and early 1880s, his wider writings on the evolutionary process in the 1880s, and his efforts later in that decade to improve the slums of Edinburgh, this essay reveals Geddes's deep, unifying, and energizing commitment to Spencer's program. Recapturing that commitment can not only help historians rethink Spencerian science as practice but also suggest ways of overcoming the dichotomy between elite ideas and practical activity that bedevils much contemporary historiography of science. PMID- 19554870 TI - Does science education need the history of science? PMID- 19554871 TI - Taxonomy and why history of science matters for science: a case study. PMID- 19554872 TI - Everything's coming up ARRA. HIM-related provisions of stimulus bill blooming as spring turns to summer. PMID- 19554873 TI - Information therapy. PMID- 19554874 TI - A call for HIM action. ARRA holds opportunity, challenge for HIM professionals. PMID- 19554875 TI - The big game. PMID- 19554877 TI - New records opening to HIM professionals. Wider content and record management initiatives offer new roles. PMID- 19554876 TI - Thinking lean in healthcare. PMID- 19554878 TI - Enabling HIE while protecting privacy. An overview of state legislative approaches. PMID- 19554879 TI - A new EHR standard for behavioral health. Stakeholders publish profile for use in behavioral health EHRs. PMID- 19554880 TI - Linking medical librarians and PHRs. PMID- 19554881 TI - HIPAA: 43,691 privacy complaints and counting. PMID- 19554882 TI - The current state of PHRs. PMID- 19554883 TI - Understanding national coverage policies. Navigating the maze of HACs, serious reportable events, and wrong surgical sites. PMID- 19554884 TI - ICD-10-CM official coding guidelines. PMID- 19554885 TI - A personal view of the personal health record. HIM professional sees the personal and professional benefits of PHRs. PMID- 19554886 TI - [Conflict of interest and adaptation]. PMID- 19554887 TI - The challenge of conflict of interest in medicine. AB - As the relationships between industry and medicine increase, the primary goals of medicine, -- high quality research, responsible patient care and excellent medical education -- are at risk of being compromised by the undue pursuit of financial gain. Conflict of interest policies and procedures are designed to reduce that risk. This article seeks to clarify the concept of conflict of interest, counter the most common objections to its use, and describe the various methods of regulating conflict of interest. PMID- 19554888 TI - Doctors and drug companies: is the dangerous liaison drawing to an end? AB - The Introduction and Overview will contain a bold statement of the essay themes. Next the section on Entanglement will look at the many forms of doctor-drug company relationships, and the evidence for how these can distort practice. Selling Sickness will briefly explore the way doctor-drug company relationships are important to the widening of boundaries of treatable illness. And finally, Disclosure to Disentanglement will look at self-regulatory and statutory attempts around the world to get more independence in the relationship between doctors and drug companies. PMID- 19554889 TI - [Conflicts of interest and clinical drug trials -- impact of the pharmaceutical industry and the impingement on the integrity of medical science]. AB - Numerous studies have addressed the subject of the extent and consequences of financial ties between pharmaceutical companies, academic institutions and clinical researchers. Based on these reports, guidelines for the management of conflicts of interest have been developed which give priority to the patients' welfare on all levels of interaction between doctors and industrial sponsors. The declaration of potential conflicts of interest and strict adherence to guidelines are mandatory to retain the physicians' credibility, reputation, and professional autonomy. Enhanced transparency in the conduct and analysis of clinical trials should be regarded as a valuable tool to uncover and avoid undue influence on clinical trials and derived prescription recommendations. PMID- 19554890 TI - [Position paper: conflict of interest in the German health industries involving the work of German International Transparency]. PMID- 19554891 TI - [Sampling studies in ambulatory quality assurance -- using the example of colonoscopy]. AB - BACKGROUND: Sampling inspections are an approved instrument for assuring and promoting the quality of healthcare. Individual documentations of medical services are requested from physicians and randomly selected for quality rating by experienced peer reviewers. For example, sampling inspections are stipulated by law for certain ambulatory services (e.g., colonoscopies) delivered by SHI authorised physicians in order to maintain professional performance standards of colonoscopists. OBJECTIVES: On behalf of the regional Association of Statutory Health Insurance Physicians (ASHIP) experienced colonoscopists regularly rate selected visual documentations (videotapes, photographs) of colonoscopies performed by SHI-authorised physicians in the ambulatory care sector. For anatomical reasons, however, a certain proportion of colonoscopies of inadequate quality will generally be tolerated. Whenever this value is exceeded, ASHIP may impose sanctions against the physician. The question is how sampling inspections have to be performed and dimensioned in order to ensure that the tests be sufficiently meaningful in terms of sensitivity and specificity. METHODS: Relevant sampling test parameters such as the false-positive rate (physicians are wrongly accused of inadequate quality) and the false-negative rate (existing deficiencies are not identified) are calculated. The calculations are performed analytically or, in the case of complex sampling test scenarios, numerically by means of computer simulations. RESULTS: The calculations show that single-stage sampling tests usually do not result in acceptable values for the false-positive and the false-negative rates. For example, a sampling test which requires the documentation of 20 colonoscopies will -- assuming some reasonable tolerance of inadequately performed colonoscopies -- result in a false-positive rate of 6% and a false-negative rate of 47%. The false-positive-rate, which is particularly relevant from a legal point of view, can be reduced by providing a two-stage sampling test. A significant reduction of the false-negative-rate may be achieved by (multiple) repetition of the single-stage sampling test and consideration of cumulative probabilities. CONCLUSIONS: In principle, sampling inspections permit statements in terms of probabilities only. In sampling inspections of healthcare quality false-negative rates are usually considered, i.e., the probability that the test is unable to identify existing quality deficiencies. However, false positive rates also need to be considered in cases where sanctions may be imposed against the physician on the basis of a positive sampling test. Numerical calculations of false-positive and false-negative rates for simple and complex sampling test scenarios should be performed in order to choose the optimum procedure and dimension of a sampling test. PMID- 19554892 TI - [The effectiveness of CME -- quality improvement through differentiated advanced medical education research]. AB - Continuing medical education (CME) increasingly focuses on measurable patient outcomes. Nevertheless, international data on this issue are insufficient, and in Germany the measurable effects of CME in terms of its efficacy and utility for patient care have hardly been subjected to scientific examination. Advanced medical education as a continuation of university education is always based on scientific standards and research-oriented learning. Advanced medical training in Germany therefore requires the implementation of a kind of learning and teaching research that should be geared toward individual training needs, personal motivation and the outcomes of medical care. In addition, the definition of educational goals and the advancement of CME in terms of continuing professional development (CPD) should be considered an important component for a reevaluation of CME. PMID- 19554893 TI - [Workplace learning for final-year medical students: a comprehensive analysis of student's expectancies and experiences]. AB - BACKGROUND: The amended German license to practice medicine (AAppO) focuses on practically oriented, patient-centred medical education. After the cancellation of the former medical doctor traineeship "Arzt im Praktikum" (AiP) qualification period in October 2004, the final year of medical education, the so-called "practical year" (PJ), has become a key teaching module in realising the demands of the AAppO. So far no studies have been carried out in the German-speaking area to qualitatively assess the specific needs and central problems that PJ students are facing. The aim of this investigation was to representatively evaluate the expectancies and experiences of final-year medical students in a workplace learning environment. METHODS: 65 students (30 male, 35 female, age 25.4 +/- 2 years) of two German university hospitals participated in focus groups both prior and subsequent (n = 60; drop-out rate 7.7%) to a 4-month period of internal medicine traineeship during their final year. Focus group protocols were transcribed and subjected to content analysis. RESULTS: 1137 individual statements on relevant topics were identified from the transcriptions. Students widely reported lack of time on the part of supervising doctors and exorbitant assignment to routine jobs. This results in insufficient training of independent patient management, followed by feelings of uncertainty and the fear of making medical errors. CONCLUSIONS: Specific educational strategies and motivation of medical teachers are important factors to the workplace learning environment. A structured curriculum including cognitive, procedural and communicative learning objectives and the introduction of defined time slots for bedside teaching is considered a good first step in this process. PMID- 19554894 TI - [After mercury, what? Planning and implementation of digital electronic thermometry]. PMID- 19554895 TI - [Burns, new challenges to take on]. AB - This article concerns a transversal descriptive study which shows the characteristics of burns treated in a Primary Health Care Center in an urban environment in Barcelona from 19 July 2005 unti 11 August 2007 (N=93). Patients younger than 15 were excluded from this study. 88% (82; CI of 95% 81,47-94,59) of the burns treated were caused by a thermal agent. Kitchen cooking oil ranks first as the cause of burns (24; 27%, CI of 95% 17,99-36,01). 70% of the burns studied had signs of superficial skin damage (65, CI of 95% 60,70-79,30). 61% (57; CI of 95% 51,70-70,30) of these burns were located on upper extremities The average recorded body surface burned was 0.0076% (median = 0,005%, range = 0,0001 0,5000%). The greatest number of wounds were observed among men aged 31 to 45 (17%; 16; CI of 95% 9,38-24,62). Educational health programs which focus on prevention of, and first aid care for, burns are needed. Studies like this one may prove useful when starting preventive or educational strategies. PMID- 19554896 TI - [Special surfaces for managing pressure in pediatrics (II). Choice, assigned algorithm (Tarise) and management models]. AB - Bed sores among children are an adverse effect provoked by the application of new technology adapted to pediatrics. Special surfaces for managing pressure in pediatrics are a preventive measure effective to avoid the development of these lesions. So that children benefit from this preventive measure, it must be adapted to their specific circumstances. In order for this to occur, it is fundamental to know: the specific characteristics which differentiate children from adults, and the type of special surfaces for managing pressure in pediatrics which are available on the market and to evaluate their appropriateness and effectiveness. The Group of Nurses to Improve Quality in Pediatrics at the University Clinical Hospital in Valencia has developed some tools which make it possible to manage and assign different sizes and types of special surfaces for managing pressure in pediatrics by means of a scientific method (Tarise). These are based on anthropometric measurements (Pediatric Space table) for each age range, the risk to develop a bed sore or skin ulcer due to pressure, the presence of a bed sore, the pathological seriousness and the type of special surfaces for managing pressure in pediatrics. PMID- 19554897 TI - [Cardiac insufficiency-failure and life quality]. AB - Cardiac insufficiency-failure is a chronic cardiovascular disease whose prevalence, incidence, mortality rate and health system costs are increasing. Patients suffering from cardiac insufficiency-failure have co-life-threatening risks which lead to severe limitations in daily activities and decrease their perception of living a quality lifestyle. Their evaluation helps to plan out future treatment and serves to predict their response to treatment. Many patients give more importance to their lifestyle quality rather than their lifespan. In this review, the authors describe the variables which affect the life quality of patients who suffer cardiac insufficiency-failure and what are the instruments more commonly employed. The authors ran a bibliographical search on Medline and Cuiden. The authors analyze five articles; they prioritize publication date, distinct magazines, study sector source of each article and the diverse profession the authors of each article has. PMID- 19554898 TI - [Risk of contracting HIV]. AB - The authors identify social activities related to the risk of contracting HIV in nursing students, bearing in mind their influence while using bio-secure methods, by means of an exploratory and qualitative study; 168 Superior School of Nursing in Lisbon, Portugal students participated in this study. Data were gathered by means of a Free Association Word Test which includes social demographic variables related to the participants. 90% of the 168 students in this study were female and 10% male. The majority 79,2%, of the participants age fell between 21 and 25 years and 97,6% of these participants were Portuguese. In the free association word test when responding to the stimulus HIV/AIDS, participants expressed disease (which is contagious in a clinical setting), and death as the awaited outcome for it. For the test term "risk of contagion when dealing with an AIDS patient" the participants often mentioned the words protection against contagion, as well as fear to contact blood and gloves as the main prevention method. To the stimulus measures of bio-security the words most frequently cited were glove as a protective measure in the context of professional activity and preservative (condom) to avoid contagion in other social contexts experienced by the participants. PMID- 19554900 TI - [Osteoporosis. Awareness and knowledge of this condition by the general public]. AB - The author provides answers for basic questions regarding osteoporosis and presents the results of a health education course about osteoporosis which took place in the "Xaloc" Health Center. PMID- 19554899 TI - [Public use of a semiautomatic external defibrillator on patients during their pediatric ages]. AB - The authors analyze the particularities of the operating algorithm in Basic Vital Support and the use of a semiautomatic external defibrillator on patients during their pediatric ages according to the latest recommendations by the ERC-2005, European Resuscitation Council. PMID- 19554901 TI - [Professional image based on written communication. Study on communicative and pragmatic aspects of nursing case files]. AB - This author compiled a 100 page descriptive study of observations on nursing practices in three hospitals. A writer's options on writing their message and a receptor's options on receiving those messages in order to infer the communicative intention of the writer are some of the elements which make it possible to gain professional meaning from the language used in nurses' case files. Physical support and its use are also variables in this analysis. The manner which we select to send a message says more than overall meaning of the words which form that message. From these messages one can gain insights into the position of a professional regarding a patient, the professional's vision and organization of his/her time and the space around the writer. PMID- 19554902 TI - [Nursing approach to ulcers and wounds. Skin anatomophysiology]. PMID- 19554903 TI - A recipe for financial disaster? Income, business sense, media, and more. PMID- 19554904 TI - Oral thrush in a one-month old infant: etiology and treatment. PMID- 19554905 TI - Iodine compounds in endodontics: an update review. PMID- 19554906 TI - The EndoVac method of endodontic irrigation: Part 4, Clinical use. PMID- 19554908 TI - Cap it? Or wrap it? PMID- 19554907 TI - Renewing the smile zone: changing our patients' lives. PMID- 19554909 TI - The importance of shade interpretation when bleaching. PMID- 19554910 TI - Upgradeable dentistry: Part 1. PMID- 19554911 TI - Fixed complete denture using implants and computer-guided technology. AB - An alternative approach to managing the completely edentulous patient with implants and a fixed complete denture has been described. A trial complete denture was fabricated, a CT scan with a double scan technique was used with specialized surgical planning software to preplan the surgery and produce an accurate stereolithography surgical guide providing all the necessary information to fabricate a cast with the desired position/angulations of the implants. This enabled the fabrication of a fixed complete denture prior to the surgical procedure with the insertion of the prosthesis immediately after the surgery was performed. PMID- 19554912 TI - Current clinical solutions for a single missing tooth: MID techniques. PMID- 19554913 TI - Real world of dentistry meets the virtual world, Part 4: Advanced techniques. PMID- 19554914 TI - Systemic diseases and dental treatment: talking with your patients. PMID- 19554915 TI - [Treatment of bronchial asthma in adults. Current advances]. AB - The last 20 years have shown an incredible improvement in the treatment of bronchial asthma, largely due to an earlier and more diffuse use of the inhaled glucocorticoids, that represent the pharmacological class of anti-asthma drugs more important to control the disease. Considering its efficacy and safety the current pharmacological treatment of asthma will probably remain unchanged for the next 10 years. PMID- 19554916 TI - [Heart without HDL.Tangier disease]. AB - A 37-year-old man was referred to our lipid clinic because of profound plasma lipid alteration. He presented large and orange tonsils, hepatosplenomegaly. No corneal opacities or other ocular abnormalities as well as no nervous system abnormalities were evident. Haematologic alterations included thrombocytopenia and stomatocytes. HDL and apolipoprotein A-I concentrations were 4 and 3.9 mg/dL, respectively. Because of a severe coronary atherosclerosis documented by coronary angiography, he underwent percutaneous revascularization. Nine months later, he experienced restenosis of the proximal anterior descending coronary artery and was referred for CABG. The patient was diagnosed with Tangier disease on the basis of the pathognomonic triad of the disease: HDL deficiency, low plasma cholesterol concentration accompanied by normal (or even elevated) triglyceride levels and hyperplastic orange tonsils. PMID- 19554917 TI - [A case of quadruple simultaneous second primary metachronous carcinomas subsequently involving colon, prostate, kidney and lung in an octogenarian patient]. AB - The authors describe a case of quadruple second primary metachronous carcinomas subsequently occurred in an octogenarian patient: colon, prostate, kidney and, lastly, lung. No chemotherapy and or radiotherapy were given. Survival resulted three years after diagnosis of the first carcinoma and four years after the last carcinoma. The particular features of the reported case are discussed with reference to literature, including the angiogenetic hypothesis. PMID- 19554918 TI - [Hospital management of acute respiratory failure: the role of the pulmonologist and of the respiratory intensive care unit]. AB - Acute respiratory failure (ARF) is one of the most common and severe urgencies of the modern medicine which may require the application of mechanical ventilation and a careful monitoring of the patient's conditions. With the popularity of non invasive ventilation and the interest of the pulmonologist for the care of the respiratory critical patient, in Italy there has been the spreading of Respiratory Intensive Care Units (RICU), which are as intermediate specialist structures in terms of intensity of care between the General Intensive Care Unit and the ordinary ward. In this article, the author analysed the cultural, scientific and organizational aspects of the central role played by the pulmonologist who's working in the RICU in the complex intra-hospital multi disciplinary management of ARF. PMID- 19554919 TI - [Transient elastography]. AB - Transient elastography (TE, Fibroscan) is a recent, rapid and highly reproducible technique for measuring liver tissue stiffness. Several studies in patients with chronic liver disease evidenced that it accurately predicts the degree of hepatic fibrosis and that it is particularly accurate in identifying severe fibrosis and cirrhosis. However, in the clinical practice TE provides an answer to a single question (liver elasticity) that could be influenced by several factors, mainly by hepatic fibrosis, but also by necrosis-inflammatory activity and extrahepatic cholestasis. PMID- 19554920 TI - [Cardiorenal syndrome, current understanding]. AB - The term cardiorenal syndrome (CRS) has increasingly been used in recent years without a constant meaning and a well accepted definition. To include the vast array of interrelated derangements, and to stress the bi-directional nature of the heart-kidney interactions, the classification of the cardiorenal syndrome includes today five sub-types whose etymology reflects the primary and secondary pathology, the time-frame and simultaneous cardiac and renal co-dysfunction secondary to systemic disease. The cardiorenal syndrome can be generally defined as a pathophysiologic disorder of the heart and kidneys whereby acute or chronic dysfunction in one organ may induce acute or chronic dysfunction in the other organ. Type I CRS reflects an abrupt worsening of cardiac function (e.g. acute cardiogenic shock or decompensated congestive heart failure) leading to acute kidney injury. Type II CRS describes chronic abnormalities in cardiac function (e.g. chronic congestive heart failure) causing progressive and permanent chronic kidney disease. Type III CRS consists in an abrupt worsening of renal function (e.g. acute kidney ischaemia or glomerulonephritis) causing acute cardiac disorder (e.g. heart failure, arrhythmia, ischemia). Type IV CRS describes a state of chronic kidney disease (e.g. chronic glomerular disease) contributing to decreased cardiac function, cardiac hypertrophy and/or increased risk of adverse cardiovascular events. Type V CRS reflects a systemic condition (e.g. diabetes mellitus, sepsis) causing both cardiac and renal dysfunction. Biomarkers can help to characterize the subtypes of the CRS and to indicate treatment initiation and effectiveness. PMID- 19554921 TI - How it works. Can we build it? PMID- 19554922 TI - Case studies. Built to last. PMID- 19554923 TI - Training. Learning on the job. PMID- 19554924 TI - Sustainability. Eco innovation. PMID- 19554925 TI - When health reform hits grandma. PMID- 19554926 TI - The Rx for good health. PMID- 19554927 TI - This doctor does not want to see you. PMID- 19554928 TI - Getting well while you shop. PMID- 19554929 TI - Staying sane may be easier than you think. PMID- 19554930 TI - Food--as pharma. PMID- 19554931 TI - A user's guide. To good health at every age. PMID- 19554932 TI - Social norms. Facebook and divorce. PMID- 19554933 TI - Federal action is required. PMID- 19554934 TI - The market can fix the problem. PMID- 19554935 TI - The state of America's health. PMID- 19554936 TI - Enlisting an army of mothers. PMID- 19554937 TI - How health bills strain budgets. PMID- 19554938 TI - Closer look at vaccines. PMID- 19554939 TI - A call for research on vaccine safety. PMID- 19554941 TI - It's never too early to be heart smart. PMID- 19554942 TI - Avoiding allergy--is it possible? PMID- 19554943 TI - The amazing teen brain. PMID- 19554945 TI - Flesh, not canvas: self-expression through body art has its drawbacks. PMID- 19554944 TI - Helping teens steer clear of trouble. PMID- 19554946 TI - Getting real about breast cancer. PMID- 19554947 TI - Secrets of the quitters. PMID- 19554948 TI - Heart disease and lung cancer aside, smokers have many reasons to quit. PMID- 19554949 TI - Weighing in for love, to keep a prenuptial vow. PMID- 19554951 TI - Ten tips for a mother-to-be. PMID- 19554950 TI - Planning ahead for a fertile future. PMID- 19554952 TI - Mad about money? Avoid a fight. PMID- 19554953 TI - Younger than your years. PMID- 19554961 TI - Steeling yourself for healthcare bills. PMID- 19554962 TI - The skinny on vitamins. PMID- 19554963 TI - Take your mark. Get set. Go. Here's what you need to do to get fit. PMID- 19554964 TI - Prescription: don't be lonely. PMID- 19554965 TI - Meditate your angst away. PMID- 19554967 TI - The time is right for health reform. PMID- 19554966 TI - Lights out! It's time for bed. PMID- 19554968 TI - The influence of improved air quality on mortality risks in Erfurt, Germany. AB - Around the world, daily variations in ambient air pollution have been consistently associated with variations in daily mortality. The aim of the study presented here was to assess the effects of ambient air pollution on daily mortality during a period of tremendous changes in air quality in the city of Erfurt, in eastern Germany, from October 1991 to March 2002. Data on particle size distributions were obtained from September 1995 to March 2002 at a research monitoring station. For particles from 0.01 microm to 2.5 microm in diameter, number concentrations (NCs)* and mass concentrations (MCs) were calculated. Particles with diameters less than or equal to 0.10 microm are defined as ultrafine particles (UFP). Data on the gaseous pollutants NO2, CO, SO2, and O3 and on PM10 (particulate matter [PM] with aerodynamic diameter less than or equal to 10 microm) were obtained from a government air-monitoring station. Data on changes in energy consumption, car fleet composition, and population were collected from local authorities. Death certificates of persons living in and dying in Erfurt were abstracted, and daily mortality counts were calculated. Poisson regression models were used to analyze the data, applying penalized splines (also known as P-splines) to model nonlinear relationships in the confounders. Model selection was done without air pollutants in the models, based on a combination of goodness-of-fit criteria and avoidance of autocorrelation in error terms. Final models included P-splines of time trend, meteorologic data, and influenza epidemics as well as day of the week with an indicator variable. Results are presented as change per interquartile range (IQR), i.e., change in the relative risk of mortality associated with a change in the concentration from the 25th to the 75th percentile of a given pollutant. Air pollutants were considered both as linear terms and as P-splines to assess the exposure-response functions. Changes in effect estimates over time were calculated using fully Bayesian time-varying coefficient models. This method was selected over four other approaches tested in simulation studies. Air-pollution concentrations decreased substantially in Erfurt during the decade under observation. The strongest changes were observed for SO2, for which annual concentrations decreased from 64 microg/m3 in 1992 to 4 microg/m3 in 2001. Concentrations of PM10, PM2.5 (particulate matter with aerodynamic diameter less than or equal to 2.5 microm), and CO decreased by more than 50%. NO2, O3, and ultrafine particles also decreased, though to a lesser extent. Based on visual inspection of the data on the changes in ambient air-pollution concentrations during the study period, we defined three study subperiods: A first subperiod from 1991 to 1995; a second, transitional subperiod from 1995 to 1998; and a third subperiod from 1998 to 2002. Generally, air-pollution concentrations decreased substantially from the first subperiod to the second, and some additional decreases occurred from the second subperiod to the third. During the second, transitional subperiod, natural gas replaced coal as the main energy source in Erfurt. In addition, the number of cars with catalytic converters increased over time, as did the number of cars in general. To facilitate the interpretation of the results, we organized the air pollutants into four groups: (1) NO2, CO, and ultrafine particles, (2) PM10 and PM2.5, (3) SO2, and (4) O3. We observed a 1.6% increased risk for daily mortality (CI, -0.4% to 3.5%) for an increase of 19.7 microg/m3 in NO2 (lag day 3), a 1.9% increased risk (CI, 0.2%-3.6%) for an increase of 0.48 mg/m3 in CO (lag day 4), and a 2.9% increased risk (CI, 0.3%-5.5%) for an increase of 9743/cm3 in ultrafine particles (lag day 4). No consistent associations were observed for PM10, PM2.5, or SO2. For O3, a 4.6% increased risk for daily mortality (CI, 1.1% 8.3%) was associated with a 43.8 microg/m3 maximum 8-hr concentration of O3 per day (lag day 2). For all four pollutants, exposure-response functions suggested no deviation from linearity. However, in time-varying models the strongest associations were observed for NO2, CO, and ultrafine particles during the transition subperiod, from 1995 to 1998, when O3 concentrations were lowest. Changes in source characteristics or ambient air-pollution concentrations were not able to explain these observations in a straightforward manner. However, the observations suggested that changes such as the introduction of three-way catalytic converters in cars and the substitution natural gas for coal might have been beneficial. Overall we concluded that: 1. Economic and political changes and the adoption of new technologies in eastern Germany resulted in distinct improvements in ambient air quality; 2. Urban air pollution in Erfurt changed within one decade from the eastern mixture toward that of western Europe ("western mixture"), which is dominated by concentrations of NOx, O3, fine particles, and ultrafine particles with low concentrations of SO2; 3. There was an association between daily mortality and ultrafine particles and combustion related gases (lag days 3 or 4); 4. Ultrafine particles seemed to be the best pollution indicator and to point to the role of local combustion in the pollution mixture; 5. Regression coefficients showed variation over time for NO2, CO, ultrafine particles, and O3 that could not be explained by nonlinearity in the exposure-response functions; 6. Mortality associated with pollution was lower at the end of the 1990s than during the 1990s, except for mortality associated with O3; and 7. Mortality associated with pollution was strongest in the second, transitional subperiod, from 1995 to 1998, when changes in source characteristics had taken place but the benefits of improved ambient air quality had not yet been completely achieved. PMID- 19554969 TI - Effects of long-term exposure to traffic-related air pollution on respiratory and cardiovascular mortality in the Netherlands: the NLCS-AIR study. AB - Evidence is increasing that long-term exposure to ambient air pollution is associated with deaths from cardiopulmonary diseases. In a 2002 pilot study, we reported clear indications that traffic-related air pollution, especially at the local scale, was related to cardiopulmonary mortality in a randomly selected subcohort of 5000 older adults participating in the ongoing Netherlands Cohort Study (NLCS) on diet and cancer. In the current study, referred to as NLCS-AIR, our objective was to obtain more precise estimates of the effects of traffic related air pollution by analyzing associations with cause-specific mortality, as well as lung cancer incidence, in the full cohort of approximately 120,000 subjects. Cohort members were 55 to 69 years of age at enrollment in 1986. Follow up was from 1987 through 1996 for mortality (17,674 deaths) and from late 1986 through 1997 for lung cancer incidence (2234 cases). Information about potential confounding variables and effect modifiers was available from the questionnaire that subjects completed at enrollment and from publicly available data (including neighborhood-scale information such as income distributions). The NLCS was designed for a case-cohort approach, which makes use of all the cases in the full cohort, while data for the random subcohort are used to estimate person-time experience in the study. Full information on confounders was available for the subjects in the random subcohort and for the emerging cases of mortality and lung cancer incidence during the follow-up period, and in NLCS-AIR we used the case cohort approach to examine the relation between exposure to air pollution and cause-specific mortality and lung cancer. We also specified a standard Cox proportional hazards model within the full cohort, for which information on potential confounding variables was much more limited. Exposure to air pollution was estimated for the subjects' home addresses at baseline in 1986. Concentrations were estimated for black smoke (a simple marker for soot) and nitrogen dioxide (NO2) as indicators of traffic-related air pollution, as well as nitric oxide (NO), sulfur dioxide (SO2), and particulate matter with aerodynamic diameter < or = 2.5 microm (PM2.5), as estimated from measurements of particulate matter with aerodynamic diameter < or = 10 microm (PM10). Overall long-term exposure concentrations were considered to be a function of air pollution contributions at regional, urban, and local scales. We used interpolation from data obtained routinely at regional stations of the National Air Quality Monitoring Network (NAQMN) to estimate the regional component of exposure at the home address. Average pollutant concentrations were estimated from NAQMN measurements for the period 1976 through 1996. Land-use regression methods were used to estimate the urban exposure component. For the local exposure component, geographic information systems (GISs) were used to generate indicators of traffic exposure that included traffic intensity on and distance to nearby roads. A major effort was made to collect traffic intensity data from individual municipalities. The exposure variables were refined considerably from those used in the pilot study, but we also analyzed the data for the full cohort in the current study using the exposure indicators of the pilot study. We analyzed the data in models with the estimated overall pollutant concentration as a single variable and with the background concentration (the sum of regional and urban components) and the local exposure estimate from traffic indicators as separate variables. In the full-cohort analyses adjusted for the limited set of confounders, estimated overall exposure concentrations of black smoke, NO2, NO, and PM2.5 were associated with mortality. For a 10-microg/m3 increase in the black smoke concentration, the relative risk (RR) (95% confidence interval [CI]) was 1.05 (1.00-1.11) for natural-cause (nonaccidental) mortality, 1.04 (0.95-1.13) for cardiovascular mortality, 1.22 (0.99-1.50) for respiratory mortality, 1.03 (0.88 1.20) for lung cancer mortality, and 1.04 (0.97-1.12) for noncardiopulmonary, non lung cancer mortality. Results were similar for NO2, NO, and PM2.5. For a 10 microg/m3 increase in PM2.5 concentration, the RR for natural-cause mortality was 1.06 (95% CI, 0.97-1.16), the same as in the results of the American Cancer Society Study reported by Pope and colleagues in 2002. The highest relative risks were found for respiratory mortality, though confidence intervals were wider for this less-frequent cause of death. No associations with mortality were found for SO2. Some of the associations between the traffic indicator variables used to assess traffic intensity near the home and mortality reached statistical significance in the full cohort. For an increase in traffic intensity of 10,000 motor vehicles in 24 hours (motor vehicles/day) on the road nearest a subject's residence, the RR was 1.03 (95% CI, 1.00-1.08) for natural-cause mortality, 1.05 (0.99-1.12) for cardiovascular mortality, 1.10 (0.95-1.26) for respiratory mortality, 1.07 (0.96-1.19) for lung cancer mortality, and 1.00 (0.94-1.06) for noncardiopulmonary, non-lung cancer mortality. Results were similar for traffic intensity in a 100-m buffer around the subject's residence and living near a major road (a road with more than 10,000 motor vehicles/day). Distance in meters to the nearest major road and traffic intensity on the nearest major road were not associated with any of the mortality outcomes. We did not find an association between cardiopulmonary mortality and living near a major road as defined using the methods of the pilot study. In the case-cohort analyses adjusted for all potential confounders, we found no associations between background air pollution and mortality. The associations between traffic intensity and mortality were weaker than in the full cohort, and confidence intervals were wider, consistent with the smaller number of subjects. The lower relative risks of mortality associated with traffic variables in the case-cohort study population could be related to the particular subcohort that was randomly selected from the full cohort, as the risks estimated with the actual subcohort were well below the average estimates obtained for 100 new case-cohort analyses with 100 alternative subcohorts of 5000 subjects each that we randomly selected from the full cohort. Differences in adjusted relative risks between the full-cohort and the case cohort analyses could be explained by random error introduced by sampling from the full cohort and by a selection effect resulting from the relatively large number of missing data for variables in the extensive confounder model used in the case-cohort analyses. More complete control for confounding probably did not contribute much to the lower relative risks in the case-cohort analyses, especially for the traffic variables, as results were similar when the limited confounder model for the full cohort was used in analyses of the subjects in the case-cohort study population. In additional analyses using black smoke concentrations as the exposure variables, we found that the association between overall black smoke and cardiopulmonary mortality was somewhat stronger for case cohort subjects who did not change residence during follow-up, and in the full cohort, there was a tendency for relative risks to be higher for subjects living in the three major cities included in the study. Adjustment for estimated exposure to traffic noise did not affect the associations of background black smoke and traffic intensity with cardiovascular mortality. There was some indication of an association between traffic noise and cardiovascular mortality only for the 1.6% of the subjects in the full cohort who were exposed to traffic noise in the highest category of > 65 A-weighted decibels (dB(A); decibels with the sound pressure scale adjusted to conform with the frequency response of the human ear). Examination of sex, smoking status, educational level, and vegetable and fruit intake as possible effect modifiers showed that for overall black smoke concentrations, associations with mortality tended to be stronger in case-cohort subjects with lower levels of education and those with low fruit intake, but differences between strata were not statistically significant. For lung cancer incidence, we found essentially no relation to exposure to NO2, black smoke, PM2.5, SO2, or several traffic indicators. Associations of overall air pollution concentrations and traffic indicator variables with lung cancer incidence were, however, found in subjects who had never smoked, with an RR of 1.47 (95% CI, 1.01 2.16) for a 10-microg/m3 increase in overall black smoke concentration. In the current study, the mortality risks associated with both background air pollution and traffic exposure variables were much smaller than the estimate previously reported in the pilot study for risk of cardiopulmonary mortality associated with living near a major road (RR, 1.95; 95% CI, 1.09-3.51). The differences are most likely due to the extension of the follow-up period in the current study and to random error in the pilot study related to sampling from the full cohort. Though relative risks were generally small in the current study, long-term average concentrations of black smoke, NO2, and PM2.5 were related to mortality, and associations of black smoke and NO2 exposure with natural-cause and respiratory mortality were statistically significant. Traffic intensity near the home was also related to natural-cause mortality. The highest relative risks associated with background air pollution and traffic variables were for respiratory mortality, though the number of deaths was smaller than for the other mortality categories. (ABSTRACT TRUNCATED) PMID- 19554970 TI - Tuberculosis case--finding and treatment in the central prison of Qazvin province, Islamic Republic of Iran. AB - As a part of a comprehensive programme of tuberculosis (TB) control in the central prison in Qazvin, Islamic Republic of Iran, a programme of active case finding was carried out from February 2004 to July 2005. From the 768 prisoners examined, 41 (5.3%) were suspected of TB and gave sputum samples. A total of 7 smear-positive TB cases were found, giving a TB prevalence in the prison of 910 per 100,000, 113 times the total TB prevalence in Qazvin province in the same year. From 7 diagnosed and treated patients, 4 were cured with the category 1 standard regimen, 1 completed treatment and 2 failed to complete treatment. Improvements are needed in TB case recognition of prisoners, especially newcomers. PMID- 19554971 TI - In vitro activity of gentamicin and amikacin against Salmonella enterica serovar Typhi: a search for a treatment regimen for typhoid fever. AB - The emergence of Salmonella enterica serovar Typhi isolates resistant to ciprofloxacin and 3rd-generation cephalosporins is a concern for physicians in developing countries. This study assessed the in vitro activity of gentamicin and amikacin against 464 S. entenca serovar Typhi isolates obtained from blood of patients clinically suspected of enteric fever who attended the Calcutta School of Tropical Medicine from 1991 to 2003. The isolates were sensitive to gentamicin and amikacin, showing minimum inhibitory concentrations 0.01-4 microg/mL and 0.005-3.5 microg/mL respectively. Both agents showed bactericidal activity at concentrations of 2 microg/mL after incubation for 6 hours. Aminoglycoside antibiotics such as gentamicin and amikacin may thus be introduced as a treatment regimen for typhoid fever. PMID- 19554973 TI - Isolation of Yersinia spp. from cases of diarrhoea in Iraqi infants and children. AB - All 250 children presenting with diarrhoea at 2 teaching hospitals in Mosul, Iraq over a 9-month period were studied for the presence of Yersinia spp. in stools by cold-enrichment culture at 4 degrees C for 21 days. Pathogenicity of the isolated Yersinia was determined. Antibodies to Y. enterocolitica were raised for rapid Yersinia detection in the stool. Yersinia spp. were isolated from the stools of only 4 patients; 3 isolates were identified as Y. enterocolitica and 1 was Y. pseudotuberculosis. The blood culture was also positive for Y. enterocolitica in 1 case. The antibiogram test for the isolated Yersinia was determined. Cross reaction between Y. pseudotuberculosis and Salmonella typhi or S. paratyphi B, and between Y. enterocolitica and Brucella was detected serologically. PMID- 19554972 TI - C-reactive protein: a useful marker for guiding duration of antibiotic therapy in suspected neonatal septicaemia? AB - The study aimed to determine whether serum C-reactive protein (CRP) levels can be used to identify when antibiotics can safely be discontinued in cases of suspected neonatal septicaemia. Neonates with suspected neonatal septicaemia treated at a hospital in Al Ramadi city, Iraq, in 2005 had serum CRP and blood cultures (the gold standard) done at admission and at 48 hours, 4 days and 6 days after starting treatment. Of the 55 neonates, CRP was < or =6 mg/L at 48 hours in 37 (67.3%) and antibiotics could be stopped in 32 (82.5%), i.e., when blood culture was negative. The moderate sensitivity (78%) and negative predictive value (86%) of serum CRP in this study suggest that this test alone cannot be used for guiding duration of antibiotic treatment for neonatal sepsis. PMID- 19554974 TI - Efficacy of diphtheria and tetanus vaccination in Gaza, Palestine. AB - This study evaluated the effectiveness and usefulness of vaccination against diphtheria and tetanus in different age groups in Gaza, Palestine. Blood samples were collected from 180 children aged <12 years, 90 males and 90 females. Using ELISA methods, the efficacy of vaccination was estimated at 87.8% for diphtheria and 98.3% for tetanus. Mean serum titres varied significantly by age group: for diphtheria 0.24 IU/mL at age 2-4 years, 0.63 IU/mL at 7-8 years and 0.46 IU/mL at 11-12 years, and for tetanus 1.01 IU/mL, 2.63 IU/mL and 1.20 IU/mL respectively. The relatively low antibody titres, especially for diphtheria, suggest the need for a booster dose. PMID- 19554975 TI - Seroprevalence of mumps before inclusion of mumps vaccination in the Iranian expanded programme on immunization. AB - The current target groups for measles, mumps and rubella vaccination in the Islamic Republic of Iran are children at 12 months and 4-6 years. A study of the age-specific seroprevalenceRepublic of Iran are children at 12 months and 4-6 years. A study of the age-specific seroprevalence of antibodies against mumps in children aged 3-18 years in Shahrekord aimed to establish the needof antibodies against mumps in children aged 3-18 years in Shahrekord aimed to establish the need for booster vaccinations to cover non-immune children. Of 338 children, 19.8% were seronegative. Age-specific seronegativity was 33.3%, 20.5% and 4.6% in age groups 7-11, 12-14 and 15-18 years respectively. To obtain herd immunity, we suggest that for the next 5 years children aged 7-11 years entering guidance school are selected as the main group for vaccination. PMID- 19554976 TI - Rationale, methods and first results of the Iranian national programme for prevention of chronic diseases from childhood: CASPIAN Study. AB - The CASPIAN Study aims to implement a school-based surveillance system for prevention of noncommunicable diseases from childhood in the Islamic Republic of Iran. The baseline survey was conducted from November 2003 to May 2004 in 23 provinces among 21,111 school students aged 6-18 years and their parents. The data collected included: birth weight and current anthropometric measurements; dietary and exercise habits; family history of chronic diseases; and knowledge about prevention. Fasting blood sugar and lipid profile were assessed in a subsample of 4811 students. PMID- 19554977 TI - Gall bladder contractility in children with beta-thalassaemia. AB - We studied gall bladder contractility in 61 children with beta-thalassaemia who were asymptomatic for gall bladder disease and 51 sex- and age-matched controls in Cairo, Egypt, using, andreal-time ultrasonography. Multiple gall bladder stones were present in 18.0% of thalassaemia patients and sludge in 6.6%. There were statistically significant differences between thalassaemia patients controls in gall bladder fasting volume, residual volume, emptying time and contraction index. There was significant positive correlation between fasting and residual volumes and age, weight and height, and between fasting volume and body mass index and serum ferritin level. Contraction index was negatively correlated with serum total bilirubin. Impaired gall bladder motility was evident in patients with beta-thalassaemia and it may be related to disease duration, serum ferritin and total serum bilirubin level. PMID- 19554978 TI - Stunting among primary-school children: a sample from Baghdad, Iraq. AB - The aim of this study was to establish baseline information about the prevalence and distribution of growth deficit (stunting) in a sample of Iraqi schoolchildren from the capital, Baghdad. A cross-sectional descriptive analysis of the growth status of 5286 primary-school children aged 7-12 years (2888 males, 2398 females) was conducted by measuring the prevalence of stunting (height-for-age Z-score < 2), and underweight (body mass index-for-age) using the NCHS/WHO and laTE/WHO cutoff values respectively. The prevalence of stunting only, and concurrent stunting and underweight, were 18.7% and 13.5% respectively, with a slight predominance among girls. The study also demonstrated the progression of height deficit with increasing age. PMID- 19554979 TI - Possible role of TORCH agents in congenital malformations in Gorgan, northern Islamic Republic of Iran. AB - This descriptive, cross-sectional study was carried out to explore the frequency of contamination with TORCH agents in neonates with congenital malformations in a referral centre in Gorgan city, Islamic Republic of Iran. Blood samples were taken from 64 neonates and their mothers over a 20-month period in 2003-04. Serologic tests showed that 4/64 infants born with congenital malformations (6%) had positive 1 gM antibody titres for Texoplasma gondil (2 cases), rubella virus (1 case) and cytomegalovirus (1 case). 1 gM was positive in 9/63 mothers (14%), also for T. gondil (3 cases), rubella virus (3 cases) and cytomegalovirus (3 cases). No cases of herpes simplex virus type II or Treponema pallidum were found. PMID- 19554980 TI - Maternal serum zinc deficiency in cases of neural tube defect in Gorgan, north Islamic Republic of Iran. AB - Previous studies have shown a high rate of neural tube defects (NTD) in Gorgan, northern (tub efcs (NTD) in Gorgan, northern Islamic Republic of Iran. This case control study during 2003-04 compared serum zinc levels and other variables in 23 mothers of neonates affected with NTD and 36 mothers with normal healthy neonates in Dezyani hospital in Gorgan. Mean serum zinc levels in the case and control groups were 13.43 micromol/L (SD 6.3) and 11.41 micromol/L (SD 6.3) respectively. Zinc deficiency was found in 13 (36.5%) of the cases and 7 (19.4%) of the controls. Logistic regression analysis showed an association between the presence of NTD and zinc deficiency (OR 5.06; 95% CI: 1.51-16.94). PMID- 19554981 TI - Sanjad-Sakati syndrome/Kenny-Caffey syndrome type 1: a study of 21 cases in Kuwait. AB - We studied 21 patients with Sanjad-Sakati syndrome (SSS) from 16 families. Parental consanguinity was recorded in 2 families (12.5%). All patients had severe intrauterine growth retardation, short stature, small hands and feet, blue sclera, deep-set eyes, microcephaly, persistent hypocalcaemia and hypoparathyroidism. Medullary stenosis was detected in 2 patients. Cytogenetic and fluorescent in situ hybridization studies were normal. All affected persons had homozygous deletion of 12 bp (155-166del) in exon 3 of the TBCE gene. All of the parents were heterozygous carriers of this mutation. The high frequency of SSS and low frequency of consanguineous marriages in this study may reflect a high rate of heterozygous carriers. PMID- 19554983 TI - Feasibility of quality of life assessment in routine clinical oncology practice: a Tunisian study. AB - Limited research has been devoted to quality of life (QOL) of cancer patients in developing countries. To assess the feasibility of QOL assessment in a cohort of Tunisian cancer patients, the EORTC QLQ-C30 questionnaire was administered to 23 women treated with adjuvant chemotherapy for early breast cancer on an outpatient basis at baseline and during the 3rd cycle of chemotherapy. We observed a significant deterioration in physical functioning, cognitive functioning and social functioning during chemotherapy. However, a wide range of methodological and practical obstacles to routine QOL evaluation were identified through this study. Further improvement of cancer care infrastructure and public education is still needed before reliable QOL studies can be performed. PMID- 19554982 TI - Development of a stress scale for pregnant women in the South Asian context: the A-Z Stress Scale. AB - Stress in pregnancy can lead to low-birth-weight and preterm babies and to psychological consequences such as anxiety and depression during pregnancy and the puerperium. Previous scales to measure stress contain items that overlap with the symptoms of pregnancy. A stress scale was developed based on in-depth interviews with pregnant women in Pakistan. Construct validity, test-retest reliability and inter-rater reliability were carried out. Cronbach alpha was 0.82 for the 30 short-listed items, with item-total correlations of 0.2-0.8. Multidimensional scaling determined 2 dimensions: socioenvironmental hassles and chronic illnesses. This was the first scale developed for pregnant women based on stressors in a developing country in South Asia. PMID- 19554984 TI - An epidemiological study on survival of oropharyngeal cancer cases in Alexandria, Egypt. AB - A retrospective cohort study was conducted to determine the 5-year survival and prognostic factors for survival for 407 oropharyngeal cancer cases registered in the 3 main hospitals in Alexandria, Egypt, from 1996-2000. Survival analysis was conducted using Kaplan-Meier curves, and multivariate Cox regression analysis. The overall 5-year survival rate was 30.8%. Multivariate analysis showed that significant prognostic factors for survival were tumour stage (hazard ratio = 2.39; 95% CI: 1.41-11.72), tumour site and patient's age. Secondary prevention of oral cancers can be conducted through examination of the oral cavity to find precancerous and early cancerous lesions and hence improve survival. PMID- 19554985 TI - Caries prevalence and treatment needs of healthy and medically compromised children at a tertiary care institution in Saudi Arabia. AB - We examined the caries prevalence and treatment needs of medically compromised and healthy paediatric patients attending the dental clinic of a tertiary care institution in Saudi Arabia. The medical records of 175 healthy and 211 medically compromised 5-year-old Saudi children were reviewed. Data was extracted on clinical status and number of decayed, extracted and filled teeth (deft score). A total of 91.9% of medically compromised and 84.0% of healthy children had evidence of caries. Mean deft score was 9.91 for medically compromised and 6.25 for healthy children. A high proportion of carious teeth (79%) were untreated, with no significant difference between the 2 groups. PMID- 19554986 TI - Knowledge and practice of university students in Lebanon regarding contraception. AB - We evaluated knowledge and practice of Lebanese university students regarding contraception in a cross-sectional, comparative study on students in public and private universities using an Arabic language self-administered questionnaire. We found low levels of knowledge of contraception. The majority of males (73.3%) and a few females (21.8%) declared previous sexual relations: the majority of males had used a condom (86.1%), but females had generally not used contraceptives (75.6%). PMID- 19554987 TI - Prevalence and predictors of white-coat hypertension in a large database of ambulatory blood pressure monitoring. AB - The objective of this study was to determine both the prevalence of white-coat effect and white-coat hypertension (WCH) and which selected clinical variables were predictors of WCH. A total of 2462 patients underwent ambulatory blood pressure monitoring either in borderline hypertension (group 1) or for assessment of antihypertensive treatment (group 2) or for hypotension (group 3). In the overall population 33.0% of patients showed WCH, 32.8% in group 1 and 37.0% in group 2. In multivariate analysis, sex and grade of hypertension were independent predictors of WCH in groups 1 and 2. PMID- 19554988 TI - Effect of interactive group discussion among physicians to promote rational prescribing. AB - This study assessed the effect of an educational intervention (interactive group discussion) on the prescribing behaviour of 51 general physicians from the north west of Tabriz. Prescriptions were analysed pre-intervention and post intervention (control and intervention groups) using a proforma with 8 indicators of correct prescribing. The mean number of drugs per prescription pre intervention was 3.82. The percentage of prescriptions with antibiotics, corticosteroids and injections were 40.8%, 25.9% and 58.0%, respectively. Following the intervention there were slight but not significant changes in the indicators in both intervention and control groups compared with pre-intervention results. PMID- 19554989 TI - Which patients wait longer to be seen and when? A waiting time study in the emergency department. AB - We investigated the waiting time for patients before seeing a physician in the emergency department of a tertiary care hospital in Riyadh, Saudi Arabia. Bi- and multivariate analyses of routine data for 2187 patients determined the association between selected patient characteristics and waiting time. The median waiting time between triage and being seen by a physician was 35.0 min (range 1.0 325.0 min). Age, day of arrival, time of arrival and triage category were significantly associated with waiting time. Older patients and those arriving on Sundays and Wednesdays waited longer. Variability in waiting times could be addressed by more standardized triage policies, but may also be influenced by other clinical or non-clinical factors that required further investigation. PMID- 19554990 TI - Opium use in a rural area of the Islamic Republic of Iran. AB - Because of its geographic proximity to the major drug production centres, there is easy access to narcotic drugs in the Islamic Republic of Iran despite efforts by governmental and nongovernmental organizations. Using a structured questionnaire as a basis for conversation, local health workers interviewed 310 residents of a rural area in Babol province about opium use. The self-reported rate of opium use, adjusted due to a bias in the sex ratio of the sample, was 8.9%. All the 42 opium users reported opium use at least 2-3 times per week in the previous 3 months. Opium was smoked by 95.2% and taken orally by 4.8%; there was no injecting use. There was no reported use of other substances, including alcohol. There was a statistically significant relationship between opium use and male sex, unemployment and cigarette smoking. PMID- 19554991 TI - Water-pipe (narguile) smokers in Lebanon: a pilot study. AB - We carried out a comparative study to assess the demographic and social characteristics of water pipe (WP) smokers, the association with cigarette smoking and chronic respiratory diseases and the dependence profile on 4 groups: exclusive WP smokers, exclusive cigarette smokers, mixed smokers and absolute non smokers. Cigarette smoking was statistically significantly higher in WP smokers than non-WP smokers; 36.5% of exclusive WP smokers smoked > or =7 WPs/week. Chronic respiratory disease and chronic bronchitis were reported more frequently in exclusive WP smokers than absolute non-smokers. WP smoking seems to be as great a risk factor as cigarette smoking for chronic respiratory disease. PMID- 19554992 TI - Injection safety at primary health care level in south-western Saudi Arabia. AB - In a study of injection safety in Abha health district, Saudi Arabia, data were collected from 47 physicians and 85 nurses at 24 primary health care centres, using an observation checklist and an interview questionnaire. All centres used individually packed disposable syringes and puncture-proof containers to collect used needles. Needlestick injury in the previous year was reported by 14.9% of physicians and 16.5% of nurses (0.21 and 0.38 injuries/person/year respectively). Logistic regression analysis identified recapping the needle after use (physicians and nurses) and bending the needle before disposal (physicians) as significant risk factors for injury. PMID- 19554993 TI - Survey of keratinophilic fungi in sewage sludge from wastewater treatment plants of Mazandaran, Islamic Republic of Iran. AB - To isolate keratinophilic fungi in sewage sludge from wastewater treatment plants in Sari city, Mazandaran province, Islamic Republic of Iran, samples were taken from 7 plants with different sewage treatment technologies. From 35 sludge samples cultured on Sabouraud's agar with cycloheximide and chloramphenicol, 326 fungal colonies belonging to 7 species were isolated. Geotrichum (59.5%), Cladosporium (13.8%), Alternaria (11.3%) and Penicillium (10.7%) species were the most prevalent. No growth of keratinophilic fungi was observed on this medium. However, using the hair-baiting technique, Microsporum gypseum, Chrysosporium spp. and Geotrichum spp. were isolated. PMID- 19554994 TI - Epidemiology of animal bites in Rafsanjan, southeast of Islamic Republic of Iran, 2003-05. AB - The incidence of animal bites in the Islamic Republic of Iran has been increasing in recent years. In a descriptive study of 1542 patients referred to a health centre in Rafsanjan city from 2003 to 2005, the incidence of animal bites in the area was estimated as 180, 195 and 241 per 100 000 in 2003, 2004 and 2005 respectively. Of the patients, 74% were bitten by dogs, 23% by cats and 3% by other animals (monkeys, rats, foxes, bats, donkeys, etc.). The mean time delay from injury to initial management was 15.1 (SD 29.8) hours. Rabies vaccine was given to 85% of cases, rabies vaccine plus rabies immunoglobulin to 15% and tetanus toxoid vaccine to 66%. PMID- 19554995 TI - Hospital-based caesarean section in the Arab region: an overview. AB - This article provides an overview of hospital-based rates of caesarean delivery in 18 Arab countries and the association between these rates and selected demographic and socioeconomic characteristics. Data on caesarean section were based on 2 of the most recent national hospital-based surveys in each country and on published studies based on hospital samples. High levels of caesarean delivery were found in Egypt (26% in 2003), followed by Sudan (20% in 1993). Six countries and the West Bank area of Palestine had rates >15% and 11 countries and Gaza had rates between 5% and 15%. The findings indicate an increasing trend of caesarean section delivery in the region. Policies aiming at reducing surgical deliveries should seek to identify and address these risk factors. PMID- 19554996 TI - Food-borne salmonella outbreak at a bank cafeteria: an investigation in an Arab country in transition. PMID- 19554998 TI - [Recognizing the importance]. PMID- 19554997 TI - [Acute pancreatitis secondary to L-asparaginase (a case report)]. PMID- 19554999 TI - [One year revised nursing care regulation--expert summary: after reform is before reform]. PMID- 19555000 TI - [Central nursing documentation supports "emotional nursing": more nursing--less bureaucracy]. PMID- 19555001 TI - [Quality assurance by professional wound documentation: often professional terminology is lacking]. PMID- 19555003 TI - [2 studies of accidental falls of patients in cardiology departments: incidence and sequelae]. PMID- 19555002 TI - [Computerized standard report for MDK (Medical Service of Public Health Insurance) evaluation of inpatient geriatric nursing: the initial overview]. PMID- 19555004 TI - [Significance of empathetic perception for nurses and patients: empathy is not pity]. PMID- 19555005 TI - [Procedures within the scope of emergency care: acute chest pain]. PMID- 19555006 TI - [Studies about use and application of nursing classifications]. AB - PROBLEM: Many nurses are not well educated in the use of nursing classifications. Shortcomings in application of nursing diagnoses and missing coherence with nursing interventions and outcomes were described in various studies. The following studies aimed to analyse the effects of implementing nursing diagnoses on nursing interventions and outcomes. METHODS: Two systematic literature reviews, instrument development and testing, and two experimental studies to implement nursing diagnostics were carried out. RESULTS: Careful implementation of nursing classifications into practice by means of educational measures resulted in accurate nursing diagnoses. Accordingly, nurses chose more effective nursing interventions that led to significantly enhanced patient outcomes. CONCLUSION: Based on the results of these studies we recommend implementation of NANDA-I nursing diagnoses with related, theory based nursing interventions and nursing outcomes into clinical practice. PMID- 19555007 TI - [Rights and responsibilities in professional relations: work and compensation responsibility: no compensation without work performance, or?]. PMID- 19555008 TI - [Prevention in nursing education--8: Thrombosis prevention: promoting venous return]. PMID- 19555009 TI - [Recognizing and understanding illnesses: "inflamed eyes" as the leading symptom]. PMID- 19555010 TI - [Internet portal for foreign practicum, employment and voluntary service: planning the step to a foreign country]. PMID- 19555011 TI - [From case to case: RAI--an instrument for assessing nursing care need]. PMID- 19555012 TI - Eyes on the swine. PMID- 19555013 TI - Blasts from the past. PMID- 19555014 TI - Hitching a ride. PMID- 19555015 TI - Juvenile thoughts. PMID- 19555016 TI - The specter of fraud. PMID- 19555017 TI - Deep into the red. PMID- 19555018 TI - Avatar acts. PMID- 19555019 TI - Act now on global warming. PMID- 19555020 TI - Still needed: a climate plan. PMID- 19555021 TI - I want to believe. PMID- 19555022 TI - From the moon to Mars. PMID- 19555023 TI - New ways to squash superbugs. PMID- 19555024 TI - Grassoline at the pump. PMID- 19555025 TI - Origins of the left & right brain. PMID- 19555026 TI - The ivory trail. PMID- 19555027 TI - The science of bubbles & busts. PMID- 19555028 TI - [Is it always necessary to use triple combination in the treatment of patients with chronic cardiac failure? Choice of the third neurohormonal blocker]. PMID- 19555029 TI - [Arterial hypertension in patients with chronic obstructive pulmonary disease in combination with coronary artery disease]. AB - AIM: To study clinicofunctional features of arterial hypertension (AH) in patients with chronic obstructive pulmonary disease (COPD) including COPD patients with coronary artery disease (CAD); 24-h profile of blood pressure (BP) and changes in intracardiac hemodynamics; to compare the disturbances detected. MATERIAL AND METHODS: 1256 patients were divided into 3 groups, 395 AH patients with COPD entered group 1; 509 AH patients with COPD and CAD, angina of effort- group 2, 352 AH patients with CAD--group 3. RESULTS: Most patients with comorbid diseases have 24-h BP profile of non dipper and night picker types in the presence of high heart rate explained by COPD effect on AH course. Lower BP in group 2 patients results from progressive maladaptive heart remodeling, attenuation of the systolic function, increasing number of patients with a restrictive type of transmitral blood flow. CONCLUSION: COPD patients with AH have more severe and frequent changes in target organs, poor prognosis as these diseases aggravate each other. PMID- 19555030 TI - [Analysis of factors affecting circulation in the territory of internal carotid artery in arterial hypertension]. AB - AIM: To assess carotid blood flow in healthy subjects and hypertensive patients, to specify prognostic significance of factors affecting carotid circulation with use of a multivariate analysis. MATERIAL AND METHODS: Clinical examination, ultrasonic dopplerography of the internal carotid arteries (ICA), echocardiography, tests for blood lipids were performed in 132 subjects (114 patients with arterial hypertension of stage I--III, 18 normotensive subjects). RESULTS: The role of age, blood pressure, blood lipids was essential both for initial and stenotic changes in ICA territory. Smoking was also an important factor. Moreover, impairment of carotid circulation correlated with progressive decline of myocardial function in the presence of coronary artery disease. PMID- 19555031 TI - [Prevalence of arterial hypertension in some subregions of Chuvashia]. AB - AIM: To study a possible link between prevalence of arterial hypertension (AH) among population and biogeochemical environmental factors of some subregions. MATERIAL AND METHODS: Epidemiological and statistical data from Public Health Ministry of the Chuvash Republic (CR) for 10 years and annual official reports on CR public health for 2001-2005 were analysed with reference to biogeochemical conditions in the CR regions. RESULTS: Cardiovascular mortality is leading among causes of death. AH accounts for 25% of deaths. Biogeochemical environmental factors have a direct impact on AH prevalence in subregions. CONCLUSION: Consideration of biogeochemical environmental factors in the study of chronic non infectious diseases, including AH, provides new information about prevalence of these diseases and help formulate recommendations for practical medicine. PMID- 19555032 TI - [Heart rate variability in hypertensive patients living in the region of Tyumen Ob]. AB - AIM: To examine heart rate variability (HRV), some biochemical and echocardiographic indices in hypertensive patients living in the Tyumen Ob region. MATERIAL AND METHODS: Registration of HRV, blood count and echocardiography were made in natural conditions of the North in 52 hypertensive patients (36 males, 16 females, age 20-50) living in Tyumen Ob region and 32 healthy controls (19 males, 13 females) matched by age and residence. RESULTS: Time and spectrum analyses of HRV indices show predominance of the sympathetic link of heart rhythm autonomic regulation in hypertensive patients. Night shifts affect the activity of the sympathetic link of heart rate autonomic regulation destroying its circadian pattern both in hypertensive and normotensive patients. CONCLUSION: Healthy persons initially experience overactivity of the regulatory systems and have the risk for cardiovascular diseases. Working at night shifts accelerates functional exhaustion. PMID- 19555034 TI - [Effects of hypotensive treatment on structural-functional properties of the vascular wall in hypertensive patients]. AB - AIM: To study structural-functional properties of the walls of large and middle size arteries and changes of these properties in the course of hypotensive therapy in hypertensive patients. MATERIAL AND METHODS: Of 609 participants of the trial (233 males and 376 females), 377 patients with essential hypertension stage I-II (mean age 46.9 +/- 7.2 years) entered the study group, 232 healthy subjects (mean age 45.7 +/- 6.9 years)--the control group. Pulse wave velocity (PWV) for vessels of elastic and muscular type was measured in all the examinees. Circadian blood pressure monitoring, echocardiography, duplex scanning of the common carotid artery were performed in hypertensive patients. 76 hypertensive patients were reexamined 6 months after treatment with amlodipin (n = 32) or fixed combination perindopril+indapamide (n = 44). RESULTS: Mean PWV appeared to be higher in hypertensive patients in whom PWV correlated with left ventricular (LV) remodeling and thickness of intima-media complex (IMC). After 6 months of hypotensive therapy PWV significantly lowered for both types of vessels studied. IMC thickness reduced by 10.1%, on the average, LV myocardial mass index--by 6.3%. CONCLUSION: The link between PWV, LV hypertrophy and IMC allows PWV use as a method of detection of cardiovascular remodeling in hypertension and for follow up of antihypertensive treatment effect on this remodeling in outpatient treatment of hypertension. PMID- 19555033 TI - [Individual chronotherapy of essential hypertension and compliance of patients with treatment scheme]. AB - AIM: To raise compliance of hypertensive patients with treatment scheme using individual chronotherapy. MATERIAL AND METHODS: Hypertensive patients were divided into two groups: the study group consisted of 225 patients, the comparison group--125 patients. The protocol comprised examination of the circadian blood pressure profile (CBPP), complex of BP indices, personal reaction to the disease (the psychodiagnostic test). RESULTS: The normal type of the circadian BP profile (dipper) was found only in 44.4% patients; 36, 120.2 and 9.3% patients were non-dippers, night-peakers and over-dippers, respectively. Only 51.6% patients had adaptive types of reaction to the disease. After half a year of individual antihypertensive treatment CBPP improved, adaptation to the disease was registered 1.5 times more often. 76% patients developed stable motivation to long-term treatment of hypertension. CONCLUSION: For adequate and safe chronotherapy of essential hypertension it is necessary to know an individual circadian rhythm of BP. Motivation to long-term pharmacotherapy is the best in patients with adaptive reaction to the disease. Education of hypertensive patients must be conducted in conditions of psychological comfort. Positive changes in life style and motivation to treatment should be promoted. PMID- 19555035 TI - [Unstable angina pectoris: outcomes of acute period and determinative factors]. AB - AIM: To follow up for 28 days outcomes of unstable angina (UA) and to determine factors affecting the disease course and negative potential of these factors. MATERIAL AND METHODS: Basing on UA clinical manifestations, 1477 patients aged 20 70 years observed in Tomsk in 2003-2004 were divided into 4 groups: group 1- patients with angina of new onset; group 2--patients with more frequent angina attacks; group 3--patients with more frequent and more severe angina attacks (longer, more painful, attacks at rest, less effective arrest with nitroglycerin); group 4--20 min and longer anginal attacks). RESULTS: On the disease day 29 UA stabilization was achieved in 1096 (74.12%) patients, UA exacerbation without acute coronary pathology (acute myocardial infarction or acute coronary insufficiency) was seen in 7 (0.47%) patients, 375 (25.41%) UA patients had acute myocardial infarction. The greatest number of acute coronary episodes (51.3%) occurred in group 1 patients. Myocardial infarction developed least frequently in group 4 patients (3.3%). CONCLUSION: Clinical symptoms of UA, ECG ischemic changes and diabetes mellitus are factors of UA unfavourable outcome. PMID- 19555036 TI - [Clinical efficacy of ivabradin and nebivolol addition in combined treatment of ischemic heart disease patients with left ventricular dysfunction]. AB - AIM: To assess ivabradin and nebivolol efficacy in combined treatment of ischemic heart disease patients with chronic heart failure (CHF) of functional class (FC) II-III by NYHA, impact of these drugs on quality of life, circadian indices of myocardial ischemia (CMI), left ventricular (LV) contractility. MATERIAL AND METHODS: A total of 92 patients with CHF of FC II-III (mean age 57.3 +/- 4.5 years) were randomized into 3 groups. Patients of group 1 (n = 30) received combined basic therapy: inhibitors of ACE, diuretics, aspirin, statins, on demand nitrates. Patients of group 2 (n = 33) received basic combined treatment plus nebivolol (nebilet, Berlin-Chemie/Menarini) in a dose 5.0 mg/day. Group 3 (n = 29) was given basic therapy plus ivabradin (coraxan) in a mean dose 7.5 mg. RESULTS: The addition of ivabradin and nebivolol to combined treatment of ischemic heart disease with LV dysfunction brought about control over heart rate (HR), improved quality of life, reduced severity of CHF, CMI, number of episodes of painful and painless ischemia. In reduced by nebivolol HR addition of ibavradin improved systolic and diastolic LV function. The analysis of HR variability in ivabradin administration showed enhancement of parasympathic activity in the vegetative balance. Administration of nebivolol produced modulated attenuation of sympathic activity. CONCLUSION: Addition of ivabradin and nebivolol to combined treatment of ischemic heart disease with LV dysfunction raises efficacy of treatment. PMID- 19555037 TI - [The role of cytokines in improvement of coronary restenosis risk stratification after endovascular stenting in patients with ischemic heart disease]. AB - AIM: To evaluate diagnostic and prognostic significance of the levels of proinflammatory cytokines (IL-1, IL-6, TNF-alpha) in improvement of stratification, i.e. determination of coronary restenosis risk in patients with ischemic heart disease 18 months after coronary artery (CA) stenting. The patients were divided into two groups: group 1 consisted of 30 patients with ischemic heart disease and symptoms of anginal recurrence, 38 patients of group 2 had no recurrent coronary insufficiency. Baseline examination of 68 patients with ischemic heart disease and their examination 6-24 months (18.1 +/- 1.9 months) after CA stenting were performed. RESULTS: High activation of the proinflammatory cytokines in patients with postinfarction cardiac dysfunction and after CA stenting is associated with a high rate of recurrent angina, deterioration of myocardial perfusion, progression of atherosclerosis in CA native bed. Restenosis of CA stents in patients after endovascular myocardial revascularization significantly more frequently correlates with elevated blood levels of IL-6 by 56.8% (p = 0.031). Patients with anginal recurrence caused by stent restenosis, progression of atherosclerosis in native CA developed high expression (10.2 58.1%) of TNF-alpha (p = 0.038) 18 months after endovascular revascularization. Repeated angioplasty is associated with multiple CA affection (k = 0.56, p = 0.004) and predilation before stenting (k = 0.3; p = 0.001). CONCLUSION: Dynamics of proinflammatory cytokines (IL-1, IL-6 and TNF-alpha) is efficient to use in complex diagnosis for better stratification of CA restenosis risk in endovascular stenting of patients with coronary artery disease. PMID- 19555038 TI - [Cardiorehabilitation of patients with stable ischemic heart disease: compliance with treatment]. AB - AIM: To study correlation between compliance with treatment (attendance of training sessions) and clinical symptoms in patients with ischemic heart disease (IHD) in the course of 12-week program of physical training. MATERIAL AND METHODS: Ergospirometry before and after training was performed in 71 patients with stable IHD (21.1% females, 78.9% males, mean age 56.5 +/- 10.0 years) for estimation of exercise intensity and efficacy. A total of 36 training sessions were conducted for 12 weeks. Compliance with treatment was assessed by attendance of training sessions. RESULTS: A direct correlation was found between the attendance and duration of IHD (r = 0.364), number of cardiovascular complications (r = 0.526), functional class of angina pectoris (r = 0.643), plasma cholesterol (r = 0.475). Negative correlation was seen between the attendance and age of the patients (r = 0.327), body mass (r = 0.311), systolic arterial pressure at rest (r = 0.756), VO+AEA-2 peak by ergospirometry before training (r = 0.543) and time of walk to the clinic (r = 0.711). No correlation was found between the attendance and gender of the patients (r = 0.107). CONCLUSION: Some clinical features of patients with stable IHD which may indicate low compliance with treatment were identified. These features should be taken into consideration for optimization of cardiorehabilitation programs based on dose-adjusted loads. PMID- 19555039 TI - [Cardiac function in secondary defect of interatrial septum and effects of its correction by Amplatzer occluder]. AB - AIM: To study with impulse-wave Doppler and impulse-wave tissue Doppler investigation left and right ventricular (LV, RV) function in two age groups of patients suffering from an interatrial septum defect (ISD) before and two years after transcatheter correction. MATERIAL AND METHODS: 34 patients (20 females and 14 males) with secondary ISD aged 5 to 67 years were divided into two groups by age. Group 1 consisted of 11 patients aged under 16 years (mean age 10.4 +/- 0.9 years), group 2--of 23 patients over 16 (mean age 31.8 +/- 2.6 years). 30 controls were matched by age. LV and RV functions were assessed by Doppler index (DI) using impulse-wave Doppler and impulse-wave tissue Doppler investigation before the defect repair and 24 hours, 1, 3, 6, 12 and 24 months after it. RESULTS: Indexed stretch-diameter of the defect in group 1 was 13.1 +/- 1.7 mm/m2, in group 2--11.7 +/- 0.8 mm/m2 (p > 0.05). Size of the occluder to body surface area was larger in group 1 patients (15.2 +/- 1.7 and 14.6 +/- 0.98 mm/m2, respectively; p > 0.05). There was initial LV and RV dysfunction in patients of both the groups. After transcatheter correction of ISD, RV DI normalized after 3 months after the defect correction in group 1, in group 2- only in 24 months. By linear RV function examination, after ISD correction DI reached control level in group 1 12 months, in group 2--24 months after surgery. LV DI in both groups did not differ from that of healthy controls 24 months after surgery. Lateral wall LV linear function examination after the defect correction discovered DI change after 1 month in group 1 (a 12% decrease; p < 0.01); control values were achieved only 2 years after surgery. In patients of group 2 DI reduced only 6 months after correction (by 14.5+ACUAOw- p +ADw- 0.01) and reached control values in 24 months. Tissue Doppler investigation of the mitral valve fibrous ring on the side of the interventricular septum after the occluder implantation showed that DI significantly decreased in group 1 in 1 month (by 9%). In group 2--in 6 months by 10% (p < 0.05). In 24 months DI achieved control values in both the groups. CONCLUSION: Patients with ISD have LV and RV dysfunction. Recovery of LV and RV functional activity after transcatheter repair of the defect with Amplatzer occluder takes place later (for 12-24 months), in children recovery is faster than in adults. PMID- 19555040 TI - [Hartil effects on structural-functional condition of the heart in patients with chronic cor pulmonale]. AB - AIM: To study effects of long-term administration of ACE inhibitor ramipril (hartil) on renin-angiotensin-aldosteron system (RAAS), dynamics of clinical symptoms of the disease and structural-functional indices of the right and left ventricle (RV, LI) in patients with chronic obstructive pulmonary disease (COPD) with decompensated chronic cor pulmonale (CCP). MATERIAL AND METHODS: X-ray examination, echocardiography (echo-CG), investigation of external respiration function were made in 45 patients with non-exacerbated COPD with CCP. The study group consisted of 25 COPD patients with CCP. They received ramipril (hartil) in a dose 2.5-5 mg. The control group of 20 patients did not receive ACE inhibitors. RESULTS: As shown by echo-CG, administration of hartil in decompensated CCP significantly improved diastolic LV and RV functions, reduced systolic and diastolic sizes of both ventricles and atria. To treatment month 12 the changes enhanced with improvement of the systolic function. Patients with decompensated CCP who had no long-term correction of RAAS exhibited deterioration of RV systolic and diastolic function, the size of their right atrium and ventricle enlarged, blood pressure in the pulmonary artery rose. CONCLUSION: Long-term administration of ACE inhibitor hartil in COPD patients with CCP in personally adjusted doses in outpatient conditions is effective and results in positive structural-functional changes of the right heart. PMID- 19555041 TI - [Clinical significance of heart rate variability in patients with systemic lupus erythematosus]. AB - AIM: To evaluate clinical significance of heart rate variability (HRV) in patients with systemic lupus erythematosus (SLE). MATERIAL AND METHODS: HRV was investigated by means of time-domain analysis of 24 hour ambulatory ECG of 122 SLE patients under 55 years of age and 32 age-matched healthy controls. In addition to clinical manifestations and activity of SLE, we assessed the presence of basic conventional cardiovascular risk factors (hypertension, smoking, body mass index, dyslipidemia), performed common carotid duplex scanning with measurement of intima-medial thickness (IMT). Inflammatory markers (ESR, CRP, IL 6) were assessed in all the patients. RESULTS: Significantly lower HRV and the trend to tachycardia were detected in SLE patients when compared to the control group. There was a significant positive correlation between HRV and a cumulative dose of cyclophosphamide, a high density lipoprotein cholesterol level, a negative correlation between HRV and cumulative dose of azathioprine, standard risk factors (hypertension, smoking, body mass index, triglyceride level), markers of inflammation (ESR, CRP, IL-6) and IMT. CONCLUSION: Measurement of HRV in combination with routine cardiovascular risk factors and level of inflammatory markers can be used for identification of subjects at risk for faster progression of atherosclerosis in SLE patients. PMID- 19555042 TI - [Pulmonary hypertension in patients with ankylosing spondilitis: main factors of development]. AB - AIM: To detect factors associated with onset of pulmonary hypertension (PH) in patients with ankylosing spondilitis (AS). MATERIAL AND METHODS: 102 patients aged 38.1 +/- 8.9 years (18-58 years) with a documented diagnosis of AS were examined with estimation of clinical idices, smoking status, spinal motility, chest excurtion, AS activity, severity of functional impairment. External respiration function was assessed at spirography, systolic pressure in the pulmonary artery (SPPA) was studied with echocardiography. PH was stated at SPPA > or = 36 mm Hg. Endothelial function was assessed by dopplerography of the brachial artery in the test with reactive hyperemia (endothelium-dependent vasodilation -EDVD) and nitroglycerin (non-endothelium-dependent vasodilation - NEDVD). As a marker of endothelial affection we used the level of circulating endothelial cells (CEC). RESULTS: PH patients were characterized by older age, body mass index, longer smoking history, AS duration, more severe functional impairment by BASFI, worse spinal motility. Both subgroups had moderate reduction of respiratory volumes. PH patients had much lower EDVD while CEC was higher. CONCLUSION: Some factors were revealed which are associated with PH onset in AS patients irrespective of ERF disorders. Endothelial dysfunction is a pathogenetic link mediating effects of these factors, first of all hypoxia, smoking and persistent systemic inflammation, on blood pressure in the system of pulmonary circulation in AS patients. PMID- 19555043 TI - [Age- and sex-related prevalence of overweight, arterial hypertension, hyperglycemia and their combinations]. AB - AIM: To study sex and age characteristics of overweight, arterial hypertension (AH) and hyperglycemia prevalence as well as prevalence of their combination. MATERIAL AND METHODS: Complex examination of Veliky Novgorod and Novgorod region 4837 residents (2081 men and 2756 women, age 20-87 years) comprised anthropometric measurements with calculation of the body mass index (Ketle index), measurements of blood pressure, fasting blood glucose, cholesterol levels, oral glucose tolerance test (GTT). RESULTS: Prevalence of overweight and obesity in males was 50.4%, in females - 63.1%. Stage I obesity occurs twice more frequently in women than in men, stage II and III obesity four and six times more frequently, respectively. 46.8% of adults suffer from AH, hyperglycemia was found in 6.03% cases. Overweight patients had metabolic carbohydrate disturbances three times more frequently than normal weight subjects. In women prevalence of overweight combination with AH was 1.5 times higher vs men (39.9 vs 26.3%); the prevalence of overweight combination with hyperglycemia was 1.7 times higher (5.5 vs 3.2%); prevalence of AH combination with hyperglycemia was 1.7 times higher (7.72 vs 2.71%). Hyperglycemia in obese persons occurred more frequently in women under 40 and men over 40 years of age. CONCLUSION: High prevalence of overweight, obesity, AH, hyperglycemia and their combinations have been revealed. This prevalence closely depended on sex and age. Prevalence of obesity and hyperglycemia, combination of overweight with AH and hyperglycemia, combination of AH with hyperglycemia is significantly higher in female subpopulation and in older cohorts. PMID- 19555045 TI - [Radionuclide diagnosis in arterial hypertension]. PMID- 19555044 TI - [Clinical variants of cardiac damage in systemic vasculitis]. PMID- 19555046 TI - [Structural functional state of cardiovascular system in hypertensive patients]. PMID- 19555047 TI - [Extracorporeal dehydration in patients with severe chronic cardiac failure. History and present-day status]. PMID- 19555049 TI - A molecular mimic of phosphorylated prolactin (S179D PRL) secreted by eukaryotic cells has a conformation with an increased positive surface charge compared to that of unmodified prolactin. AB - S179D prolactin (S179D PRL) is a pseudophosphorylated form of human PRL which has potent antitumor and anti-angiogenic activities in vivo. This molecule binds to the same forms of the PRL receptor (PRLR) as unmodified PRL, yet this binding results in different intracellular signaling and biological end points. Since it is now clear that PRLRs are predimerized and therefore that ligand binding must initiate signaling by inducing a conformational change in the receptor dimer, we hypothesized that S179D PRL had an altered conformation compared to unmodified PRL. The conformation of the ligand-receptor ternary complex would therefore also have an altered conformation, and thus, different signaling molecules would be activated. Here we present evidence in support of this hypothesis by demonstrating, in contrast to unmodified PRL, that S179D PRL has reduced nickel and zinc binding capacity and a higher affinity for heparin and DEAE. Conformational changes have occurred since these features are counterintuitive on the basis of the simple substitution of a serine with a negatively charged aspartate residue. To demonstrate that these particular properties of S179D PRL were not due to misfolding of the molecule during production, S179D PRL was expressed in two different mammalian cell lines. Also investigated was the potential for production of S179D PRL as a soluble cytoplasmic, or secreted periplasmic, protein in Escherichia coli. PMID- 19555048 TI - [Anemia as complication of chronic heart failure: prevalence, treatment, prognosis]. PMID- 19555050 TI - Self-assembled monolayers of bisphosphonates: influence of side chain steric hindrance. AB - Bisphosphonates form self-assembled monolayers (SAMs) spontaneously on stainless steel, silicon, and titanium oxidized surfaces. We used contact angle measurements, atomic force microscopy, and X-ray reflectivity analysis to study the formation of SAMs on a model surface of ultraflat titanium (rms = 0.2 nm). The results were extended to standard materials (mechanically polished titanium, stainless steel, and silicon) and showed that water-soluble bisphosphonic perfluoropolyether can easily form SAMs, with 100% surface coverage and a layer thickness of less than 3 nm. Hydrophobic (water contact angle >110 degrees on stainless steel or titanium) and lipophobic (methylene iodide contact angle >105 degrees on titanium) properties are discussed in terms of industrial applications. PMID- 19555051 TI - Quantitative microplate-based respirometry with correction for oxygen diffusion. AB - Respirometry using modified cell culture microplates offers an increase in throughput and a decrease in biological material required for each assay. Plate based respirometers are susceptible to a range of diffusion phenomena; as O(2) is consumed by the specimen, atmospheric O(2) leaks into the measurement volume. Oxygen also dissolves in and diffuses passively through the polystyrene commonly used as a microplate material. Consequently the walls of such respirometer chambers are not just permeable to O(2) but also store substantial amounts of gas. O(2) flux between the walls and the measurement volume biases the measured oxygen consumption rate depending on the actual [O(2)] gradient. We describe a compartment model-based correction algorithm to deconvolute the biological oxygen consumption rate from the measured [O(2)]. We optimize the algorithm to work with the Seahorse XF24 extracellular flux analyzer. The correction algorithm is biologically validated using mouse cortical synaptosomes and liver mitochondria attached to XF24 V7 cell culture microplates, and by comparison to classical Clark electrode oxygraph measurements. The algorithm increases the useful range of oxygen consumption rates, the temporal resolution, and durations of measurements. The algorithm is presented in a general format and is therefore applicable to other respirometer systems. PMID- 19555053 TI - Organocatalytic highly enantio- and diastereoselective Mannich reaction of beta ketoesters with N-Boc-aldimines. AB - The catalytic enantioselective Mannich reaction promoted by chiral bifunctional organocatalysts is described. The treatment of beta-ketoesters with N-Boc aldimines under mild reaction conditions afforded the corresponding beta-amino beta-ketoesters with excellent diastereoselectivities (up to 100:0 dr) and excellent enantioselectivities (up to 99% ee). PMID- 19555052 TI - Analysis of samples stored as individual plugs in a capillary by electrospray ionization mass spectrometry. AB - Droplets or plugs within multiphase microfluidic systems have rapidly gained interest as a way to manipulate samples and chemical reactions on the femtoliter to microliter scale. Chemical analysis of the plugs remains a challenge. We have discovered that nanoliter plugs of sample separated by air or oil can be analyzed by electrospray ionization mass spectrometry when pumped directly into a fused silica nanospray emitter tip. Using leucine-enkephalin in methanol and 1% acetic acid in water (50:50 v:v) as a model sample, we found carry-over between plugs was <0.1% and relative standard deviation of signal for a series of plugs was 3%. Detection limits were 1 nM. Sample analysis rates of 0.8 Hz were achieved by pumping 13 nL samples separated by 3 mm long air gaps in a 75 microm inner diameter tube. Analysis rates were limited by the scan time of the ion trap mass spectrometer. The system provides a robust, rapid, and information-rich method for chemical analysis of sample in segmented flow systems. PMID- 19555054 TI - Influence of the solvent on the self-assembly of a modified amyloid beta peptide fragment. I. Morphological investigation. AB - The solvent-induced transition between self-assembled structures formed by the peptide AAKLVFF is studied via electron microscopy, light scattering, and spectroscopic techniques. The peptide is based on a core fragment of the amyloid beta-peptide, KLVFF, extended by two alanine residues. AAKLVFF exhibits distinct structures of twisted fibrils in water or nanotubes in methanol. For intermediate water/methanol compositions, these structures are disrupted and replaced by wide filamentous tapes that appear to be lateral aggregates of thin protofilaments. The orientation of the beta-strands in the twisted tapes or nanotubes can be deduced from X-ray diffraction on aligned stalks, as well as FT-IR experiments in transmission compared to attenuated total reflection. Strands are aligned perpendicular to the axis of the twisted fibrils or the nanotubes. The results are interpreted in light of recent results on the effect of competitive hydrogen bonding upon self-assembly in soft materials in water/methanol mixtures. PMID- 19555055 TI - Alkylaluminum derivatives of diphenic acid: novel aluminum carboxylates. AB - The reaction of trialkylaluminum compounds with diphenic acid in a 2:1 molar ratio results in the formation of novel dialkylaluminum dicarboxylates [Et(4)Al(2)(OOCC(12)H(8)COO)](3) (1) and [(i)Bu(4)Al(2)(OOCC(12)H(8)COO)](2) (2). Molecular structures of the compounds have been determined by X-ray crystallography. Compound 1 is a hexamer with a skeleton framework consisting of four fused heterocyclic rings, one 27-membered and three Al(2)O(4)C(2) 8 membered. Compound 2 is a centrosymmetric tetramer with a skeleton framework consisting of three fused heterocyclic rings, one 18-membered and two Al(2)O(4)C(2) 8-membered. The structures of the compounds reveal R(4)Al(2)O(4)C(2) subunits, each linked through diphenyl C(12)H(8) bridges. Each of the aluminum atoms is bonded to two alkyl groups and two oxygen atoms originating from two different dicarboxylate moieties. PMID- 19555056 TI - Catching CO2 in a bowl. AB - Increased concentrations of CO(2) in the atmosphere contribute to global climate change. Improved methods are needed for removing CO(2) from the flue gas of power plants and/or directly from the atmosphere. A macrocyclic amidourea recently synthesized by Brooks et al., when dissolved in DMSO along with tetrabutyl ammonium fluoride, removes CO(2) from the atmosphere to form a complex in which a CO(3) group is held by a number of O-H-N bonds within the bowl-shaped cavity of the macrocycle. We have calculated the structure, stability, and vibrational spectra of this complex, using density functional techniques and polarized double zeta basis sets. Both basis set superposition errors and polarizable continuum effects on the complex geometry and stability have been evaluated. The calculated structure is in good agreement with experiment. We predict that this CO(3)(-2) complex (and its HCO(3)(-) analogue) have larger formation constants by several orders of magnitude than the analogue complex of the amidourea macrocycle with Cl(-) (particularly in DMSO solution compared to aqueous solution). Our calculations also indicate that the CO(3)(-2) and HCO(3)(-) complexes can be distinguished by (13)C NMR. The CO(3)(-2) complex also has a distinctive H-N stretch, perturbed by the H-bonding to the CO(3) group. We also calculate the CO(3)(-2) complex to absorb within the visible region, unlike the free macrocycle or typical metal carbonates. Macrocycles of this type may provide a useful route to the absorption of atmospheric CO(2). Our calculations also indicate that changing the solvent from DMSO to water and/or heating the complex will be an efficient way to decompose it to release CO(2). PMID- 19555057 TI - Photon-initiated homolysis of peroxynitrous acid. AB - Laser flash photolysis of ONOOH at 355 nm and a pH of 4.0-5.5 causes homolysis of ONOOH nearly exclusively at the N-O bond rather than at the O-O bond (HO(2)(*)/HO(*) > 25:1). All of the NO(*) and HO(2)(*) radicals formed by photolysis subsequently recombine with a rate constant of (1.2 +/- 0.2) x 10(10) M(-1) s(-1) via second-order kinetics, as demonstrated by the return of the UV/vis absorbance to initial levels. Excitation at 266 nm also yields HO(2)(*) and NO(*), but after recombination, the absorbance levels are lower than initial values, possibly because HO(*) produced by the photolysis of water reacts with ONOOH. When NO(3)(-), the product of the ONOOH isomerization, is photolyzed, the ONOO(-) formed is rapidly protonated with a second-order rate constant of (1.7 +/ 0.8) x 10(10) M(-1) s(-1). The ONOOH decays to the starting material, NO(3)(-), with a first-order rate constant of 1.2 s(-1). The quantum yield for the photon initiated homolysis is 15% for both ONOOH and ONOO(-). We conclude that the ON OOH and ON-OO(-) bond dissociation energies are similar. PMID- 19555058 TI - Asymmetric synthesis of bis(tertiary arsines): highly stereoselective alkylations of diastereomers of a chiral phosphine-stabilized bis(arsenium triflate). AB - The addition of organolithium reagents to an equilibrating mixture of diastereomers of a phosphine-stabilized 1,2-ethanediylbis(phenylarsenium triflate) containing chiral arsenic stereocenters and an enantiomerically pure, atropisomeric tertiary phosphepine derived from lithiated (aR)-2,2'-dimethyl-1,1' binaphthalene generates unequal mixtures of diastereomers and enantiomers of chelating 1,2-ethanediylbis(tertiary arsines), chiral at arsenic, with liberation of the (aR(P))-phosphepine. Thus, the addition of methyllithium in diethyl ether at -95 degrees C to a dichloromethane solution of the complex (R*(As),R*(As))-(+/ )/(R*(As),S*(As))-1,2-[(R(3)P)PhAsCH(2)CH(2)AsPh(PR(3))](OTf)(2), where R(3)P is (aR(P))-[2-(methoxymethyl)phenyl]phosphepine, generates (R*(As),R*(As))-(+/-)-1,2 ethanediylbis(methylphenylarsine) in 78% diastereoselectivity and 95% enantioselectivity in favor of the (R(As),R(As)) enantiomer. Under similar conditions, the addition of n-butyllithium in hexanes to a solution of the bis(phosphepine-stabilized)-diarsenium triflate at -95 degrees C gives the corresponding (R*(As),R*(As))-(+/-)-1,2-ethanediylbis[(n-butyl)phenylarsine) in 77% diastereoselectivity and 93% enantioselectivity in favor of the (R(As),R(As)) enantiomer. PMID- 19555059 TI - Surprisingly bright near-infrared luminescence and short radiative lifetimes of ytterbium in hetero-binuclear Yb-Na chelates. AB - New heterobinuclear lanthanide complexes with benzoxazole-substituted 8 hydroxyquinolines, [Ln(ligand)(2)(mu-ligand)(2)Na] (Ln: Yb, Lu), have been prepared and their structure established by X-ray crystallography, (1)H NMR spectroscopy, and photophysical studies. The complexes display efficient ligand sensitized near-infrared luminescence of ytterbium at 925-1075 nm with lifetimes and quantum yields as high as 22 micros and 3.7%, in the solid state, and 20 micros and 2.6% in CH(2)Cl(2) solution, respectively. These quantum yields are the highest reported to date for ytterbium complexes with organic ligands containing C-H bonds. A long-wavelength and intense intraligand charge-transfer transition (lambda(max) = 446-456 nm; epsilon approximately 1.2 x 10(4) M(-1) cm( 1)) allows for the excitation of infrared luminescence with visible light up to 600 nm. Remarkable features of these complexes include (i) quantitative ligand-to Yb(III) energy transfer resulting in high overall efficiency of the ytterbium luminescence, (ii) unusually short radiative lifetime of the Yb(III) ion, 706-745 micros for solutions in CH(2)Cl(2), calculated from the f-f absorption spectra, and 513-635 micros estimated for solid state samples from quantum yield and lifetime data, and (iii) the unexpected large influence of second-sphere composition on the radiative lifetime of ytterbium. PMID- 19555061 TI - Nucleophile generation via decarboxylation: asymmetric construction of contiguous trisubstituted and quaternary stereocenters through a Cu(I)-catalyzed decarboxylative Mannich-type reaction. AB - The first catalytic asymmetric decarboxylative Mannich-type reaction between aldimines and cyanocarboxylic acids was developed. Alpha,alpha,beta trisubstituted beta-amino nitriles containing contiguous trisubstituted and all carbon quaternary stereocenters were produced with moderate to high enantio- and diastereoselectivity using 5 mol % CuOAc-(R)-DTBM-SEGPHOS complex as an asymmetric catalyst. This reaction proceeds through Cu(I)-catalyzed decarboxylative nucleophile generation followed by the addition of the resulting chiral Cu ketenimide to imines. Because the reaction proceeds under very mild conditions at nearly neutral pH, the reaction is applicable to a wide range of substrate combinations, including both aromatic and aliphatic imines. The products were converted to beta(2,2,3)-amino acid derivatives through simple acidic hydrolysis without any racemization or epimerization. Therefore, this is a general catalytic asymmetric method for the synthesis of beta(2,2,3)-amino acid derivatives, a group of important chiral building blocks for pharmaceutical lead compounds. PMID- 19555060 TI - Oxidation unzipping of stable nanographenes into joint spin-rich fragments. AB - When an all-benzenoid nanographene is linearly unzipped into oxygen-joined fragments, the oxidized benzenoid rings (aromatic sextets) selectively adopt the low-spin (DeltaS = 0) or high-spin conformation (DeltaS = 1) to yield the thermally most stable isomer. The selection of the conformation depends simply on the position of the aromatic sextets: the inner ones prefer the high-spin conformation, whereas the peripheral ones prefer the low-spin conformation. Therefore, the resulting most stable isomer has a total spin whose value equals the number of inner aromatic sextets (n(i)) along the oxidizing line. The nanographene fragments contained in this isomer have a ferromagnetic spin coupling. Due to the tautomerization between the high-spin and low-spin conformations, there also exist other possible isomers with higher energies and with spins at ground state ranging from 0 to (n(i) - 1). The rich geometrically correlated spins and the adjustable energy gaps indicate great potential of the graphene oxides in spintronic devices. PMID- 19555062 TI - Modeling the multivalent recognition between dendritic molecules and DNA: understanding how ligand "sacrifice" and screening can enhance binding. AB - This paper reports the application of molecular dynamics methods to understand the interactions between dendritic molecules with spermine surface groups and double-helical DNA. Importantly, we are able to reproduce the binding effects observed experimentally, indicating that this type of modeling is robust and reliable. The energetic effects were deconvoluted in order to quantify the binding of each spermine unit to the DNA double helix. Importantly, for the first generation dendron G1, DNA binding was adversely affected by increasing levels of NaCl (>10% of the interaction energy is lost). For second-generation G2 however, we observed a compensation effect, in which some ligands "sacrifice" themselves, losing large amounts of binding energy with DNA. However, these ligands screen the complex, which enables the other spermine residues to bind more effectively to DNA. In this way, the multivalent array is able to maintain its high affinity binding, even as the salt concentration increases (only ca. 1% of the interaction energy is lost). These modeling studies are in agreement with, and provide a unique insight into, the experimental results. Clearly, ligand flexibility and ability to reorganize the interactions with DNA are important, demonstrating that high levels of preorganization and ligand framework rigidity are not always beneficial for multivalent recognition. The concept suggested by this modeling study, in which ligand "sacrifice" and binding site screening combine to enable high-affinity binding, is a new paradigm in multivalency. PMID- 19555064 TI - Possible origin of electronic effects in Rh(I)-catalyzed enantioselective hydrogenation. AB - Reducing the electron density of ligands switches the regioselectivity of Rh(I) catalyzed hydrometalation. A reversal of the sense of chiral induction was also observed when chiral ligands are electronically tuned in the same manner. The combined data provide an alternative rationale for the electronic effects often observed in asymmetric hydrogenation. PMID- 19555065 TI - Observation of optical spin symmetry breaking in nanoapertures. AB - Observation of a spin symmetry breaking effect in plasmonic nanoscale structures due to spin-orbit interaction is presented. We demonstrate a nanoplasmonic structure which exhibits a crucial role of an angular momentum (AM) selection rule in a light-surface plasmon scattering process. In our experiment, the intrinsic AM (spin) of the incident radiation is coupled to the extrinsic momentum (orbital AM) of the surface plasmons via spin-orbit interaction. Due to this effect, we achieved a spin-controlled enhanced transmission through a coaxial nanoaperture. PMID- 19555063 TI - Evolution of proteins with genetically encoded "chemical warheads". AB - We recently developed a phage-based system for the evolution of proteins in bacteria with expanded amino acid genetic codes. Here we demonstrate that the unnatural amino acid p-boronophenylalanine (BF) confers a selective advantage in the evolution of glycan-binding proteins. We show that an unbiased library of naive antibodies with NNK-randomized V(H) CDR3 loops converges upon mutants containing BF when placed under selection for binding to a model acyclic amino sugar. This work represents a first step in the evolution of carbohydrate-binding proteins that use a reactive unnatural amino acid "warhead" and demonstrates that a "synthetic" genetic code can confer a selective advantage by increasing the number of functional groups available to evolution. PMID- 19555066 TI - Spin gapless semiconductor-metal-half-metal properties in nitrogen-doped zigzag graphene nanoribbons. AB - The geometries, formation energies, and electronic and magnetic properties of N doping defects, including single atom substitution and pyridine- and pyrrole-like substructures in zigzag graphene nanoribbons (ZGNRs), were investigated by means of spin-unrestricted density functional theory computations. The edge carbon atoms are more easily substituted with N atoms, and three-nitrogen vacancy (3NV) defect and four-nitrogen divacancy (4ND) defect also prefer the ribbon edge. Single N atom substitution and pyridine- and pyrrole-like N-doping defects can all break the degeneracy of the spin polarization of pristine ZGNRs. One single N atom substitution makes the antiferromagnetic semiconducting ZGNRs into spin gapless semiconductors, while double edge substitution transforms N-doped graphenes into metals. Pyridine- and pyrrole-like N-doping defects make ZGNRs into half-metals or spin gapless semiconductors. These results suggest the potential applications of N-doped ZGNRs in nanoelectronics. PMID- 19555067 TI - Palladium-catalyzed direct alkynylation of C-H bonds in benzenes. AB - Palladium-catalyzed ortho-alkynylation of aromatic C-H bonds in anilides is described. Preliminary mechanistic studies reveal that electrophilic palladation is involved. Synthetic elaborations of alkynylated products are also demonstrated. PMID- 19555068 TI - Optical two-dimensional fourier transform spectroscopy of semiconductor quantum wells. AB - Coherent light-matter interactions of direct-gap semiconductor nanostructures provide a great test system for fundamental research into quantum electronics and many-body physics. The understanding gained from studying these interactions can facilitate the design of optoelectronic devices. Recently, we have used optical two-dimensional Fourier-transform spectroscopy to explore coherent light-matter interactions in semiconductor quantum wells. Using three laser pulses to generate a four-wave-mixing signal, we acquire spectra by tracking the phase of the signal with respect to two time axes and then Fourier transforming them. In this Account, we show several two-dimensional projections and demonstrate techniques to isolate different contributions to the coherent response of semiconductors. The low-temperature spectrum of semiconductor quantum wells is dominated by excitons, which are electron-hole pairs bound through Coulombic interactions. Excitons are sensitive to their electronic and structural environment, which influences their optical resonance energies and line widths. In near perfect quantum wells, a small fluctuation of the quantum well thickness leads to spatial localization of the center-of-mass wave function of the excitons and inhomogeneous broadening of the optical resonance. The inhomogeneous broadening often masks the homogeneous line widths associated with the scattering of the excitons. In addition to forming excitons, Coulombic correlations also form excitonic molecules, called biexcitons. Therefore, the coherent response of the quantum wells encompasses the intra-action and interaction of both excitons and biexcitons in the presence of inhomogeneous broadening. Transient four-wave mixing studies combined with microscopic theories have determined that many-body interactions dominate the strong coherent response from quantum wells. Although the numerous competing interactions cannot be easily separated in either the spectral or temporal domains, they can be separated using two-dimensional Fourier transform spectroscopy. The most common two-dimensional Fourier spectra are S(I)(omega(tau),T,omega(t)) in which the second time period is held fixed. The result is a spectrum that unfolds congested one-dimensional spectra, separates excitonic pathways, and shows which excitons are coherently coupled. This method also separates the biexciton contributions and isolates the homogeneous and inhomogeneous line widths. For semiconductor excitons, the line shape in the real part of the spectrum is sensitive to the many-body interactions, which we can suppress by exploiting polarization selection rules. In an alternative two dimensional projection, S(I)(tau,omega(Tau),omega(t)), the nonradiative Raman coherent interactions are isolated. Finally, we show S(III)(tau,omega(Tau),omega(t)) spectra that isolate the two-quantum coherences associated with the biexciton. These spectra reveal previously unobserved many body correlations. PMID- 19555069 TI - Titan's ionic species: theoretical treatment of N2H+ and related ions. AB - We use different ab initio methods to compute the three-dimensional potential energy surface (3D-PES) of the ground state of N(2)H(+). This includes the standard coupled cluster, the complete active space self-consistent field, the internally contacted multi reference configuration interaction, and the newly developed CCSD(T)-F12 methods. For the description of H and N atoms, several basis sets are tested. Then, we incorporate the 3D-PES analytical representations into variational calculations of the rovibrational spectrum of N(2)H(+)(X(1)Sigma(+)) up to 7200 cm(-1) above the zero point vibrational energy. Our data show that the CCSD(T)-F12/aug-cc-pVTZ approach represents a compromise for good description of the PES and computation cost. This technique is recommended for full dimensional PES generation of atmospheric and astrophysical relevant polyatomic systems. We applied this method to derive the rovibrational spectra of N(2)H(+)(X(1)Sigma(+)) and of N(2)H(++)(X(2)Sigma(+)). Finally, we discuss the existence of the N(2)H(++)(X(2)Sigma(+)) in Titan's atmosphere. PMID- 19555070 TI - Soluble polymer carriers for the treatment of cancer: the importance of molecular architecture. AB - Chemotherapy can destroy tumors and arrest cancer progress. Unfortunately, severe side effects (treatment is usually a series of injections of highly toxic drugs) often restrict the frequency and size of dosages, much to the detriment of tumor inhibition. Most chemotherapeutic drugs have pharmacokinetic profiles with tremendous potential for improvement. Water-soluble polymers offer the potential to increase drug circulation time, improve drug solubility, prolong drug residence time in a tumor, and reduce toxicity. Cytotoxic drugs that are covalently attached to water-soluble polymers via reversible linkages more effectively target tumor tissue than the drugs alone. Macromolecules passively target solid tumor tissue through a combination of reduced renal clearance and exploitation of the enhanced permeation and retention (EPR) effect, which prevails for fast-growing tumors. Effective drug delivery involves a balance between (i) elimination of the polymeric drug conjugate from the bloodstream by the kidneys, liver, and other organs and (ii) movement of the drug out of the blood vasculature and into the tumor (that is, extravasation). Polymers are eliminated in the kidney by filtration through pores with a size comparable to the hydrodynamic diameter of the polymer; in contrast, the openings in the blood vessel structures that traverse tumors are an order of magnitude greater than the diameter of the polymer. Thus, features that may broadly be grouped as the "molecular architecture" of the polymer, such as its hydrodynamic volume (or molecular weight), molecular conformation, chain flexibility, branching, and location of the attached drug, can greatly impact elimination of the polymer from the body through the kidney but have a much smaller effect on the extravasation of the polymer into the tumor. Molecular architecture can in theory be adjusted to assert essentially independent control over elimination and extravasation. Understanding how molecular architecture affects passage of a polymer through a pore is therefore essential for designing polymer drug carriers that are effective in passively delivering a drug payload while conforming to the requirement that the polymers must eventually be eliminated from the body. In this Account, we discuss examples from in vivo studies that demonstrate how polymer architectural features impact the renal filtration of a polymer as well as tumor penetration and tumor accumulation. In brief, features that inhibit passage of a polymer through a pore, such as higher molecular weight, decreased flexibility, and an increased number of polymer chain ends, help prevent elimination of the polymer by the kidneys and can improve blood circulation times and tumor accumulation, thus improving therapeutic effectiveness. PMID- 19555071 TI - PARACEST properties of a dinuclear neodymium(III) complex bound to DNA or carbonate. AB - A dinuclear Nd(III) macrocyclic complex of 1 (1,4-bis[1-(4,7,10 tris(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane]-p-xylene) and mononuclear complexes of 1,4,7-tris-1,4,7,10-tetraazacyclododecane, 2, and 1,4,7-tris[(N-N diethyl)carbamoylmethyl]-1,4,7,10-tetraazacyclododecane, 3, are prepared. Complexes of 1 and 2 give rise to a PARACEST (paramagnetic chemical exchange saturation transfer) peak from exchangeable amide protons that resonate approximately 12 ppm downfield from the bulk water proton resonance. The dinuclear Nd(III) complex is promising as a PARACEST contrast agent for MRI applications, because it has an optimal pH of 7.5 and the rate constant for amide proton exchange (2700 s(-1)) is nearly as large as it can be within slow exchange conditions with bulk water. Dinuclear Ln(2)(1) complexes (Ln(III) = Nd(III), Eu(III)) bind tightly to anionic ligands including carbonate, diethyl phosphate, and DNA. The CEST amide peak of Nd(2)(1) is enhanced by certain DNA sequences that contain hairpin loops, but decreases in the presence of diethyl phosphate or carbonate. Direct excitation luminescence studies of Eu(2)(1) show that double stranded and hairpin-loop DNA sequences displace one water ligand on each Eu(III) center. DNA displaces carbonate ion despite the low dissociation constant for the Eu(2)(1) carbonate complex (K(d) = 15 microM). Enhancement of the CEST effect of a lanthanide complex by binding to DNA is a promising step toward the preparation of PARACEST agents containing DNA scaffolds. PMID- 19555072 TI - A new approach in the preparation of dendrimer-based bifunctional diethylenetriaminepentaacetic acid MR contrast agent derivatives. AB - In this paper, we report a new method to prepare and characterize a contrast agent based on a fourth-generation (G4) polyamidoamine (PAMAM) dendrimer conjugated to the gadolinium complex of the bifunctional diethylenetriamine pentaacetic acid derivative (1B4M-DTPA). The method involves preforming the metal ligand chelate in alcohol prior to conjugation to the dendrimer. The dendrimer based agent was purified by a Sephadex G-25 column and characterized by elemental analysis. The analysis and SE-HPLC data gave a chelate to dendrimer ratio of 30:1 suggesting conjugation at approximately every other amine terminal on the dendrimer. Molar relaxivity of the agent measured at pH 7.4 displayed a higher value than that of the analogous G4 dendrimer based agent prepared by the postmetal incorporation method (r(1) = 26.9 vs 13.9 mM(-1) s(-1) at 3 T and 22 degrees C). This is hypothesized to be due to the higher hydrophobicity of this conjugate and the lack of available charged carboxylate groups from noncomplexed free ligands that might coordinate to the metal and thus also reduce water exchange sites. Additionally, the distribution populations of compounds that result from the postmetal incorporation route are eliminated from the current product simplifying characterization as quality control issues pertaining to the production of such agents for clinical use as MR contrast agents. In vivo imaging in mice showed a reasonably fast clearance (t(1/2) = 24 min) suggesting a viable agent for use in clinical application. PMID- 19555074 TI - Fabrication of highly reflective and conductive double-surface-silvered layers embedded on polymeric films through all-wet process at room temperature. AB - An easy technique is developed to fabricate highly conductive and reflective double-surface-silvered polyimide films at room temperature by the incorporation of silver ions in surface-modified polyimide, and subsequently by the in situ reduction of silver ions in alkaline containing aqueous glucose solution. Surface properties of the silvered composite films were investigated as a function of treatment time and reducing environment, respectively. Sheet reflectivity and conductivity can be controlled by adjusting the potassium hydroxide (KOH) etching and reducing conditions. The excellent silver-polymer adhesive property is based on a "tree roots" like micro/nanostructure of the silver layers. The essential mechanical properties of the silvered films were maintained as their inside matrix is intact during the whole procedure. Different properties between one film's double-side surfaces were investigated during the fabricating process. Films were characterized by inductively coupled plasma (ICP), X-ray diffraction (XRD), contact angle (CA), scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force microscopy (AFM), four point probe instrument, and ultraviolet (UV) spectrophotometer. PMID- 19555075 TI - Structure and malonyl CoA-ACP transacylase binding of streptomyces coelicolor fatty acid synthase acyl carrier protein. AB - Malonylation of an acyl carrier protein (ACP) by malonyl Coenzyme A-ACP transacylase (MCAT) is fundamental to bacterial fatty acid biosynthesis. Here, we report the structure of the Steptomyces coelicolor (Sc) fatty acid synthase (FAS) ACP and studies of its binding to MCAT. The carrier protein adopts an alpha helical bundle structure common to other known carrier proteins. The Sc FAS ACP shows close structural homology with other fatty acid ACPs and less similarity with Sc actinorhodin (act) polyketide synthase (PKS) ACP where the orientation of helix I differs. NMR experiments were used to map the binding of ACP to MCAT. This data suggests that Sc FAS ACP interacts with MCAT through the negatively charged helix II of ACP, consistent with proposed models for ACP recognition by other FAS enzymes. Differential roles for residues at the interface are demonstrated using site-directed mutagenesis and in vitro assays. MCAT has been suggested, moreover, to participate in bacterial polyketide synthesis in vivo. We demonstrate that the affinity of the polyketide synthase ACP for MCAT is lower than that of the FAS ACP. Mutagenesis of homologous helix II residues on the polyketide synthase ACP suggests that the PKS ACP may bind to MCAT in a different manner than the FAS counterpart. PMID- 19555073 TI - Self-organization in coordination-driven self-assembly. AB - Self-assembly allows for the preparation of highly complex molecular and supramolecular systems from relatively simple starting materials. Typically, self assembled supramolecules are constructed by combining complementary pairs of two highly symmetric molecular components, thus limiting the chances of forming unwanted side products. Combining asymmetric molecular components or multiple complementary sets of molecules in one complex mixture can produce myriad different ordered and disordered supramolecular assemblies. Alternatively, spontaneous self-organization phenomena can promote the formation of specific product(s) out of a collection of multiple possibilities. Self-organization processes are common throughout much of nature and are especially common in biological systems. Recently, researchers have studied self-organized self assembly in purely synthetic systems. This Account describes our investigations of self-organization in the coordination-driven self-assembly of platinum(II) based metallosupramolecules. The modularity of the coordination-driven approach to self-assembly has allowed us to systematically study a wide variety of different factors that can control the extent of supramolecular self organization. In particular, we have evaluated the effects of the symmetry and polarity of ambidentate donor subunits, differences in geometrical parameters (e.g., the size, angularity, and dimensionality) of Pt(II)-based acceptors and organic donors, the influence of temperature and solvent, and the effects of intermolecular steric interactions and hydrophobic interactions on self organization. Our studies have shown that the extent of self-organization in the coordination-driven self-assembly of both 2D polygons and 3D polyhedra ranges from no organization (a statistical mixture of multiple products) to amplified organization (wherein a particular product or products are favored over others) and all the way to the absolute self-organization of discrete supramolecular assemblies. In many cases, inputs such as dipolar interactions, steric interactions, and differences in the geometric parameters of subunits, used either alone or as multiple factors simultaneously, can achieve absolute self organization of discrete supramolecules. We have also observed instances where self-organization is not absolute and varies in its deviation from statistical results. Steric interactions are particularly useful control factors for driving such amplified self-organization because they can be subtly tuned through small structural variations. Having the ability to fully understand and control the self-organization of complex mixtures into specific synthetic supramolecules can provide a better understanding of analogous processes in biological systems. Furthermore, self-organization may allow for the facile synthesis of complex multifunctional, multicomponent systems from simply mixing a collection of much simpler, judiciously designed individual molecular components. PMID- 19555077 TI - Conjugation of biomolecules with magnetic protein microspheres for the assay of early biomarkers associated with acute myocardial infarction. AB - This study demonstrates an improved magnetic protein microsphere-aided sandwich fluoroimmunoassay for the analysis of myoglobin and heart-type fatty acid binding protein (H-FABP), early protein markers associated with acute myocardial infarction. In preparation for the assay we constructed superparamagnetic human serum albumin (HSA)/gamma-Fe(2)O(3) microspheres, and grafted capture antibodies (monoclonal antimyoglobin 7C3 and anti-H-FABP 10E1) onto the protein microspheres using the avidin-biotin system. Then the antibody-carrying microspheres were used in a sequential sandwich fluoroimmunoassay along with detection antibodies (Alexa fluor594-labeled antimyoglobin 4E2 and FITC-labeled anti-H-FABP 9F3). The magnetic HSA/gamma-Fe(2)O(3) microspheres were characterized by scanning electron microscopy, atomic force microscopy, Fourier transform infrared spectrophotometry, atomic absorption spectrophotometry, and vibrating sample magnetometry. Fluorescence images of the post-immunoassay microspheres recorded using an inverted fluorescence microscope showed that the average fluorescence intensity was correlated with the concentration of cardiac markers, in agreement with the results obtained by an F-4500 FL spectrophotometer; this indicated that the fluoroimmunoassay could be used to semiquantitatively detect both myoglobin and H-FABP. The detection limit was 10 ng/mL for myoglobin and 1 ng/mL for H FABP. PMID- 19555078 TI - Evaluation of a high-intensity focused ultrasound-immobilized trypsin digestion and 18O-labeling method for quantitative proteomics. AB - A new method that uses immobilized trypsin concomitant with ultrasonic irradiation results in ultrarapid digestion and more thorough (18)O labeling for quantitative protein comparisons. The method was reproducible and provided effective digestions within <1 min with lower amounts of enzyme, compared to traditional methods. This method was demonstrated for digestion of both simple and complex protein mixtures, including bovine serum albumin, a global proteome extract from the bacteria Shewanella oneidensis, and mouse plasma, as well as (18)O labeling of complex protein mixtures, validating this method for differential proteomic measurements. This approach is simple, reproducible, cost effective, rapid, and well suited for automation. PMID- 19555079 TI - Development of a new device for ultrasensitive electrochemiluminescence microscopy imaging. AB - Electrochemiluminescence (ECL) is widely used in biosensors and immunoassays thanks to the high sensitivity and specificity of the electrochemically triggered luminescence signal. So far, no applications have been reported on the use of ECL as a probe for ultrasensitive low-light microscope imaging. This work reports the development of a new transparent electrochemical cell for ECL imaging suitable for single cell analysis. The system is based on the use of a microscope placed in a dark box equipped with a CCD camera and a potentiostat. Transparent conducting glass coated with fluorine-doped tin oxide (FTO) has been used, and a three electrode configuration has been designed. The electrochemical cell was optimized using 8 microm diameter polystyrene beads coated with a Ru(bpy)(3)(2+) complex in order to simulate living cells. The Ru(bpy)(3)(2+) immobilized on the microbeads can be imaged and quantified at a concentration as low as 1 x 10(-19) mol/microm(2). Microscope imaging showed that the ECL signal was detected only in correspondence to the beads present on the electrode surface, and the probe could be accurately localized with a spatial resolution of 0.4 microm. The new ECL imaging device can be used in conjunction with other chemiluminescence-based imaging methods for ultrasensitive multiplex imaging on cells and tissues. PMID- 19555080 TI - Surface-enhanced Raman scattering studies on the interaction of phosphonate derivatives of imidazole, thiazole, and pyridine with a silver electrode in aqueous solution. AB - Surface-enhanced Raman scattering (SERS) spectra from phosphonate derivatives of N-heterocyclic aromatic compounds immobilized on an electrochemically roughened silver electrode surface are reported and compared to Raman spectra of the corresponding solid species. The tested compounds contain imidazole [ImMeP ([hydroxy-(1H-imidazol-5-yl)-methyl]-phosphonic acid) and (ImMe)2P (bis[hydroxy (1H-imidazol-4-yl)-methyl]-phosphinic acid)]; thiazole [BAThMeP ((butylamino thiazol-2-yl-methyl)-phosphonic acid) and BzAThMeP ((benzylamino-thiazol-2-yl methyl)-phosphonic acid)]; and pyridine ((PyMe)2P (bis[(hydroxy-pyridin-3-yl methyl)]-phosphinic acid) aromatic rings. Changes in wavenumber, broadness, and the enhancement of N-heterocyclic aromatic ring bands upon adsorption are consistent with the adsorption primarily occurring through the N lone pair of electrons with the ring arranged in a largely edge-on manner for ImMeP and BzAThMeP or in a slightly inclined orientation to the silver electrode surface at an intermediate angle from the surface normal for (ImMe)2P, BAThMeP, and (PyMe)2P. A strong enhancement of a roughly 1500 cm(-1) SERS signal for ImMeP and (PyMe)2P is also observed. This phenomenon is attributed to the formation of a localized C=C bond, which is accompanied by a decrease in the ring-surface pi electrons' overlap. In addition, more intense SERS bands due to the benzene ring in BzAThMeP are observed than those observed for the thiazole ring, which suggests a preferential adsorption of benzene. Some interaction of a phosphonate unit is also suggested but with moderate strength between biomolecules. The strength of the P=O coordination to the silver electrode is highest for ImMeP but lowest for BzAThMeP. For all studied biomolecules, the contribution of the structural components to their ability to interact with their receptors was correlated with the SERS patterns. PMID- 19555081 TI - High-resolution temperature-concentration diagram of alpha-synuclein conformation obtained from a single Forster resonance energy transfer image in a microfluidic device. AB - We present a microfluidic device for rapid and efficient determination of protein conformations in a range of medium conditions and temperatures. The device generates orthogonal gradients of concentration and temperature in an interrogation area that fits into the field of view of an objective lens with a numerical aperture of 0.45. A single Forster resonance energy transfer (FRET) image of the interrogation area containing a dual-labeled protein provides a 100 x 100 point map of the FRET efficiency that corresponds to a diagram of protein conformations in the coordinates of temperature and medium conditions. The device is used to explore the conformations of alpha-synuclein, an intrinsically disordered protein linked to Parkinson's and Alzheimer's diseases, in the presence of a binding partner, the lipid-mimetic sodium dodecyl sulfate (SDS). The experiment provides a diagram of conformations of alpha-synuclein with 10,000 individual data points in a range of 21-47 degrees C and 0-2.5 mM SDS. The diagram is consistent with previous reports but also reveals new conformational transitions that would be difficult to detect with conventional techniques. The microfluidic device can potentially be used to study other biomolecular and soft matter systems. PMID- 19555082 TI - Micro-Raman spectroscopic observation of water expulsion induced destruction of hydrophobic clusters in crystalline lysozyme. AB - In this paper, three kinds of solid lysozyme samples with different water contents were investigated by confocal Raman spectroscopy. For the rod-like lysozyme crystal with highest water content, a sudden decrease of the intensity ratio of the doublet at 1338 and 1360 cm(-1) was observed when the ambient relative humidity (RH) was lower than 86%, indicating the destruction of hydrophobic clusters of lysozyme induced by the expulsion of the hydration water from the crystal. In contrast to the rod-like crystal, tetragonal crystal and floor-like precipitate with a smaller amount of water showed no change of the structures of the hydrophobic clusters when the relative humidity was decreasing. The presence of bulk water in the rod-like crystal is believed a necessary factor for the function of the hydration water which promotes the hydrophobicity of hydrophobic clusters. PMID- 19555083 TI - Imidazolium-based ionic liquids. 1-methyl imidazolium nitrate: thermochemical measurements and ab initio calculations. AB - In this work data of the molar enthalpies of formation of the ionic liquid 1 methylimidazolium nitrate [H-MIM][NO3] was measured by means of combustion calorimetry. The molar enthalpy of fusion of [H-MIM][NO3] was measured using DSC. Experiments to vaporize the ionic liquid into vacuum or nitrogen stream in order to obtain vaporization enthalpy have been performed. Ab initio calculations of the enthalpy of formation in the gaseous phase have been performed for the ionic species using the G3MP2 theory. The combination of traditional combustion calorimertry with modern high-level ab initio calculations allow the determination of the molar enthalpy of vaporization of the ionic liquid under study. The ab initio calculations indicate that [H-MIM][NO3] is most probably separated into the neutral species methyl-imidazole and HNO3 in the gaseous phase at conditions of the vaporization experiments. PMID- 19555084 TI - Antiproliferative bistramides from Trididemnum cyclops from Madagascar (1). AB - Bioassay-guided fractionation of a marine extract from Trididemnum cyclops afforded the new lipopeptide 39-oxobistramide K (1) and the known bistramides A (2) and D (3). Structure elucidation of 1 was carried out by analysis of one- and two-dimensional NMR spectroscopy and HRMS data. Bistramides have been reported to exhibit antiproliferative activity in the nanomolar range against a number of tumor cell lines in vitro and in vivo. The isolate 1 was tested for antiproliferative activity against the A2780 cell line and exhibited an IC(50) value of 0.34 microM. PMID- 19555085 TI - Thermally activated superradiance and intersystem crossing in the water-soluble chlorophyll binding protein. AB - The crystal structure of the class IIb water-soluble chlorophyll binding protein (WSCP) from Lepidium virginicum is used to model linear absorption and circular dichroism spectra as well as excited state decay times of class IIa WSCP from cauliflower reconstituted with chlorophyll (Chl) a and Chl b. The close agreement between theory and experiment suggests that both types of WSCP share a common Chl binding motif, where the opening angle between pigment planes in class IIa WSCP should not differ by more than 10 degrees from that in class IIb. The experimentally observed (Schmitt et al. J. Phys. Chem. B 2008, 112, 13951) decrease in excited state lifetime of Chl a homodimers with increasing temperature is fully explained by thermally activated superradiance via the upper exciton state of the dimer. Whereas a temperature-independent intersystem crossing (ISC) rate is inferred for WSCP containing Chl a homodimers, that of WSCP with Chl b homodimers is found to increase above 100 K. Our quantum chemical/electrostatic calculations suggest that a thermally activated ISC via an excited triplet state T4 is responsible for the latter temperature dependence. PMID- 19555086 TI - Pressure-dependent hydrogen permeability extended for metal membranes not obeying the square-root law. AB - Hydrogen permeability of metal membranes is generally defined by the square-root law, as the proportional coefficient of permeation flux to the square-root difference of the pressures on both sides of the membrane. However, deviation from the law has been widely reported for palladium, niobium, etc. Although n-th power instead of the square root has often been employed to determine permeability for these membranes, it has no theoretical base. These approaches do not consider concentration dependency of hydrogen diffusivity in the membrane. This study theoretically extended the definition of permeability by taking it into account, where square root of pressure was used throughout. The resultant permeability depended on pressure. This approach had the following four characteristics. First, the permeability could be qualitatively linked with pressure-dependent solution and diffusion coefficients. For this purpose, the solution coefficient was also extended from Sieverts' law. Second, the permeability could be easily evaluated from permeation flux dependent on feed side pressure, usually measured in membrane study. Third, this approach enabled comparison of permeation ability irrespective of obeying permeation law. Fourth, permeation flux could be estimated for any pressure conditions visually and analytically. Thus, analytically estimated values were more precise than those using the conventional square-root law. These characteristics are successfully demonstrated using experimental results obtained not only for a palladium membrane in this study but also for palladium and niobium membranes in the literature. PMID- 19555088 TI - Luminescent boron-contained ladder-type pi-conjugated compounds. AB - Four diboron-contained ladder-type pi-conjugated compounds 1-4 were designed and synthesized. Their thermal, photophysical, electrochemical properties, as well as density functional theory calculations, were fully investigated. The single crystals of compounds 1 and 3 were grown, and their crystal structures were determined by X-ray diffraction analysis. Both compounds have a ladder-type pi conjugated framework. Compounds 1 and 2 possess high thermal stabilities, moderate solid-state fluorescence quantum yields, as well as stable redox properties, indicating that they are possible candidates for emitters and charge transporting materials in electroluminescent (EL) devices. The double-layer device with the configuration of [ITO/NPB (40 nm)/1 or 2 (70 nm)/LiF (0.5 nm)/Al (200 nm)] exhibited good EL performance with the maximum brightness exceeding 8000 cd/m(2). PMID- 19555087 TI - Structure-based design, synthesis, and biochemical and pharmacological characterization of novel salvinorin A analogues as active state probes of the kappa-opioid receptor. AB - Salvinorin A, the most potent naturally occurring hallucinogen, has attracted an increasing amount of attention since the kappa-opioid receptor (KOR) was identified as its principal molecular target by us [Roth, B. L., et al. (2002) Proc. Natl. Acad. Sci. U.S.A. 99, 11934-11939]. Here we report the design, synthesis, and biochemical characterization of novel, irreversible, salvinorin A derived ligands suitable as active state probes of the KOR. On the basis of prior substituted cysteine accessibility and molecular modeling studies, C315(7.38) was chosen as a potential anchoring point for covalent labeling of salvinorin A derived ligands. Automated docking of a series of potential covalently bound ligands suggested that either a haloacetate moiety or other similar electrophilic groups could irreversibly bind with C315(7.38). 22-Thiocyanatosalvinorin A (RB 64) and 22-chlorosalvinorin A (RB-48) were both found to be extraordinarily potent and selective KOR agonists in vitro and in vivo. As predicted on the basis of molecular modeling studies, RB-64 induced wash-resistant inhibition of binding with a strict requirement for a free cysteine in or near the binding pocket. Mass spectrometry (MS) studies utilizing synthetic KOR peptides and RB-64 supported the hypothesis that the anchoring residue was C315(7.38) and suggested one biochemical mechanism for covalent binding. These studies provide direct evidence of the presence of a free cysteine in the agonist-bound state of the KOR and provide novel insights into the mechanism by which salvinorin A binds to and activates the KOR. PMID- 19555089 TI - Palladium(II)-catalyzed cyclization of unsaturated hydroperoxides for the synthesis of 1,2-dioxanes. AB - The cyclization of gamma,delta-unsaturated tertiary hydroperoxides in the presence of a palladium(II) catalyst afforded 1,2-dioxanes resembling biologically active natural products. A variety of substrates were screened, and synthetic manipulations were accomplished to construct compounds with structural similarity to antimalarial targets. PMID- 19555090 TI - The B-H...H-P dihydrogen bonding in ion pair complexes [(CF(3))(3)BH(-)][HPH(3 n)(Me)(n)(+)] (n = 0-3) and its implication in H(2) elimination and activation reactions. AB - The B-H(delta-)...(delta+)H-P dihydrogen bonding (DHB) in ion pair complexes [(CF(3))(3)BH(-)][HPH(3-n)(Me)(n)(+)] (n = 0-3) and its role in the combination of proton and hydride with the release of H(2) or, reversibly, the heterolytic activation of H(2) by Lewis pairs (CF(3))(3)BPH(3-n)(Me)(n) have been theoretically investigated at the MP2 and DFT levels. It is found that the B H...H-P bonds behave similarly to those in neutral pairs and ion-molecule complexes in most respects, such as the linearity of the H...H-P moiety, the characteristics of the electron transfer and rearrangement, and the topological properties of the DHB critical point, except that in certain cases, a blue shifting of the H-bond vibrational frequency is observed. In [(CF(3))(3)BH( )][HPH(3-n)(Me)(n)(+)], the proton shifting within the complexes leads to the formation of the dihydrogen complex B(CF(3))(3)(eta(2)-H(2)), which is followed by a subsequent H(2) release. The stability of B(CF(3))(3)(eta(2)-H(2)) (D(e)/D(0) = 10.8/6.0 kcal/mol) makes the proton-hydride combination proceed in a fashion similar to the protonation reactions in transition-metal hydrides rather than those in group 13 hydrides EH(4)(-) (E = B, Al, Ga). As for the H(2) splitting reaction R(3)BPR'(3) + H(2) --> [R(3)BH(-)][HPR'(3)(+)], classical Lewis pair (CLP) (CF(3))(3)BPH(3) exhibits a high barrier and results in an unstable ion pair product [(CF(3))(3)BH(-)][HPH(3)(+)] compared with the "frustrated Lewis pair" (FLP) (C(6)F(5))(3)BP(tBu)(3). A detailed analysis of the mechanistic aspects of H(2) activation by (CF(3))(3)BPH(3) and (C(6)F(5))(3)BP(tBu)(3), supported by another CLP (CF(3))(3)BP(tBu)(3) which has a binding energy comparable to (CF(3))(3)BPH(3) but a reaction exothermicity comparable to (C(6)F(5))(3)BP(tBu)(3), allows us to suggest that the low stability of FLP (C(6)F(5))(3)BP(tBu)(3) is the determining factor for the low reaction barrier. The relative stability and other properties of the ion pair products [R(3)BH(-)][HPR'(3)(+)] have also been analyzed. Results strongly support the view proposed by Rokob et al. [ Rokob , T. A. ; Hamza , A. ; Stirling , A. ; Soos , T. ; Papai , I. Angew. Chem., Int. Ed. 2008 , 47 , 2435 ] that the frustration energy lowers the energy barrier and increases the exothermicity of the reaction. PMID- 19555091 TI - On the emerging role of chemistry in the fashioning of biologics: synthesis of a bidomainal fucosyl GM1-based vaccine for the treatment of small cell lung cancer. AB - The synthesis of the novel small cell lung cancer (SCLC) fucosyl GM1-based vaccine construct, featuring insertion of the HLA-DR binding 15 amino acid sequence derived from Plasmodium falciparum, is described. The resultant glycopeptide has been synthesized in an efficient manner. Finally, successful conjugation of the glycopeptide to the keyhole limpet hemocyanin (KLH) carrier protein completed the preparation of the vaccine. PMID- 19555092 TI - Kinetic and thermodynamic barriers to chlorine transfer between amines in aqueous solution. AB - Third-order rate constants for the acid-catalyzed reversible reaction of N chlorotaurine with benzylamine and dimethylamine were determined in water at 25 degrees C and I = 0.5 (NaClO4). The reaction with benzylamine shows inverse solvent deuterium isotope effects of kH/kD=0.57 and 0.47 in the forward and reverse directions, respectively. These isotope effects, together with the absence of detectable general acid catalysis for this reaction, provide evidence for a stepwise mechanism involving fast equilibrium protonation of N chlorotaurine followed by rate-determining chlorine transfer from the protonated chloramine to benzylamine. The observation of strong catalysis by general acids of the reaction of dimethylamine with N-chlorotaurine suggests a change to a concerted mechanism with proton and chlorine transfer occurring in a single step. This change in mechanism is enforced by the absence of a significant lifetime for protonated chlorotaurine in contact with this strongly nucleophilic amine. The kinetic and thermodynamic parameters for the reaction between protonated chlorotaurine and benzylamine are used to estimate a Marcus intrinsic reaction barrier of deltaG(0)+/+ =4.1 kcal/mol for chlorine transfer between amines. Comparison of this intrinsic barrier with those reported previously for bromine transfer between carbanions points to the existence of certain similarities between halogen and proton transfer reactions. PMID- 19555093 TI - P(i-PrNCH2CH2)3N as a Lewis base catalyst for the synthesis of beta hydroxynitriles using TMSAN. AB - Proazaphosphatrane 1a was found to be an efficient catalyst for synthesis of beta hydroxynitriles via the reaction of trimethylsilylacetonitrile (TMSAN) with aldehydes under mild reaction conditions and typically low catalyst loading (ca. 2 mol %). A variety of functional groups were tolerated, and good to excellent product yields were obtained. PMID- 19555095 TI - Chemoenzymatic synthesis of rivastigmine based on lipase-catalyzed processes. AB - A straightforward chemoenzymatic synthesis of enantiomerically pure rivastigmine has been efficiently carried out under mild reaction conditions, with Candida antarctica lipase B responsible for the stereoselective acetylation of the corresponding (R)-alcohol or amine. An exhaustive enzymatic study has been developed exploring the possibilities of carry out enzyme recycling, scaling up the enzymatic process and development of a dynamic kinetic resolution procedure for the production of adequate enantiomerically pure precursors of rivastigmine. Total chemoenzymatic synthesis of this pharmaceutical has been performed in good overall yield from commercially available 3-methoxyacetophenone. PMID- 19555094 TI - Preparation of pentafluorosulfanyl (SF5) pyrrole carboxylic acid esters. AB - Pyrrole derivatives bearing a pentafluorosulfanyl group are currently unknown. In this paper, a facile preparation of SF5-substituted pyrrole carboxylic acid esters in good yield is reported. Utilizing the cycloaddition of an azomethine ylide to pentafluorosulfanylalkynes, a series of dihydropyrroles were prepared and oxidized to the respective 1-tert-butyl-4-(pentafluorosulfanyl)pyrrole-2 carboxylic acid esters in good yield. Further treatment of these pyrroles with catalytic triflic acid allowed removal of the tert-butyl group. PMID- 19555097 TI - Atom substitution effects of [XF6]- in ionic liquids. 2. Theoretical study. AB - Following the preceding spectroscopic study, we further investigate atomic mass effects of [XF6]- in 1-butyl-3-methylimidazolium cation ([BMIm]+) based ionic liquids (ILs) on dynamical natures by a computational approach in this study. We carry out the molecular dynamics simulations for 1-butyl-3-methylimidazolium cation based ILs ([BMIm][PF6], [BMIm][AsF6], and [BMIm][SbF6]) with the development of the force fields of [AsF6]- and [SbF6]- by an ab initio calculation. We have calculated density of state (DOS) and velocity autocorrelation function (VACF) profiles, polarizability time correlation function (TCF) and Kerr spectra, intermediate scattering functions, and dynamical structure factors. The decomposition analysis has been also carried out to understand the ion species and types of motion. From these computational studies, we find that the contribution of the reorientation of cations and anions mainly governs the Kerr spectrum profile in all three ILs, while the contribution of the collision-induced and cross terms, which are related to translational motions including coupling with librational motion, is not large at higher frequencies than 50 cm(-1). It is suggested that, with the atom substitution effects of anion units on interionic interactions, many properties in ILs are controllable. In addition, it is emphasized in this study that atomic mass effects in ILs are accessible through a complementary approach of both experimental and theoretical approaches. PMID- 19555098 TI - Searching the conformational space of cyclic beta-amino acid peptides. AB - There is an increasing interest in the secondary structure of beta-amino-acid containing peptides, since these compounds exhibit an intrinsic propensity to form stable folds even for short peptides, a feature that is rarely observed in alpha-amino-acid-containing peptides. In this work, we use a multiple trajectory molecular dynamics approach to study a panel of cyclic beta-amino-acid-containing peptides with a variety of motifs that differ in the ring size, ring substituents, and terminal protecting groups. We find a reasonable agreement between the predicted and the experimentally observed structures, in spite of the simple solvent representation used, indicating that in most cases the folding proceeds energetically downhill and it is driven to a great extent by structural preferences coded in the internal degrees of freedom, a result supported by our energy partition analysis. Our results also show that when the N-terminal end is unprotected, it is likely to be charged in a protic polar solvent. In that case, we find that only a molecular dynamics simulation with an "all atom" solvent representation is capable of reproducing the experimentally observed secondary structure of the peptide. Finally, the time evolution analysis of the hydrogen bond-induced turns as well as of the root-mean-square deviation from the observed structure indicates that some peptides could have a higher intrinsic flexibility than others, within a given fold, a result that correlates to some degree with our molecular mechanics energy analysis. PMID- 19555099 TI - Atom substitution effects of [XF6]- in ionic liquids. 1. Experimental study. AB - We have investigated the interionic vibrational dynamics of 1-butyl-3 methylimidazolium cation ([BMIm]+) based ionic liquids with the anions of [PF6]-, [AsF6]-, and [SbF6]- as well as the static physical properties, such as shear viscosity and liquid density. Shear viscosity for the ionic liquids becomes lower with the heavier atom anion: [BMIm][PF6]>[BMIm][AsF6]>[BMIm][SbF6]. This tendency for heavy atom substitution for the anion results from weaker interionic interaction caused by larger anion volume. Femtosecond optically heterodyne detected Raman-induced Kerr effect spectroscopy has been used to observe the interionic vibrational dynamics of ionic liquids. The interionic vibration in the frequency region of less than 50 cm(-1) clearly shows the heavy atom substitution effect; that is, the heavy atom substitution of [XF6]- critically affects the interaction-induced motion. The forthcoming paper will further provide the molecular-level insights of the heavy atom substitution effect of the [XF6]- anion on the interionic dynamics and interaction for the three ionic liquids by a molecular dynamics simulation approach. PMID- 19555100 TI - Chiral-auxiliary-mediated asymmetric synthesis of tris-heteroleptic ruthenium polypyridyl complexes. AB - A strategy for the asymmetric synthesis of chiral-at-metal [Ru(pp)(pp')(pp'')](2+) complexes, where pp, pp', and pp'' are achiral 2,2' bipyridines, is introduced. The method employs isopropyl-2-(2' hydroxyphenyl)oxazolines as chiral auxiliaries, which serve in their deprotonated forms as strong bidentate ligands that provide excellent asymmetric induction in the course of the coordination chemistry and, importantly, can afterward become substituted with complete retention of configuration in the presence of acid. PMID- 19555101 TI - Light-driven release from polymeric microcapsules functionalized with bacteriorhodopsin. AB - Bacteriorhodopsin was incorporated into the shell of polymeric capsules. Light driven variation of intercapsule volume pH with successive pore opening was demonstrated by scanning electron microscopy. Release of the encapsulated dye molecules was studied by confocal fluorescence microscopy. PMID- 19555102 TI - Locus-specific microemulsion catalysts for sulfur mustard (HD) chemical warfare agent decontamination. AB - The rates of catalytic oxidative decontamination of the chemical warfare agent (CWA) sulfur mustard (HD, bis(2-chlororethyl) sulfide) and a range (chloroethyl) sulfide simulants of variable lipophilicity have been examined using a hydrogen peroxide-based microemulsion system. SANS (small-angle neutron scattering), SAXS (small-angle X-ray scattering), PGSE-NMR (pulsed-gradient spin-echo NMR), fluorescence quenching, and electrospray mass spectroscopy (ESI-MS) were implemented to examine the distribution of HD, its simulants, and their oxidation/hydrolysis products in a model oil-in-water microemulsion. These measurements not only present a means of interpreting decontamination rates but also a rationale for the design of oxidation catalysts for these toxic materials. Here we show that by localizing manganese-Schiff base catalysts at the oil droplet-water interface or within the droplet core, a range of (chloroethyl) sulfides, including HD, spanning some 7 orders of octanol-water partition coefficient (K(ow)), may be oxidized with equal efficacy using dilute (5 wt. % of aqueous phase) hydrogen peroxide as a noncorrosive, environmentally benign oxidant (e.g., t(1/2) (HD) approximately 18 s, (2-chloroethyl phenyl sulfide, C(6)H(5)SCH(2)CH(2)Cl) approximately 15 s, (thiodiglycol, S(CH(2)CH(2)OH)(2)) approximately 19 s {20 degrees C}). Our observations demonstrate that by programming catalyst lipophilicity to colocalize catalyst and substrate, the inherent compartmentalization of the microemulsion can be exploited to achieve enhanced rates of reaction or to exert control over product selectivity. A combination of SANS, ESI-MS and fluorescence quenching measurements indicate that the enhanced catalytic activity is due to the locus of the catalyst and not a result of partial hydrolysis of the substrate. PMID- 19555103 TI - Free radical polymers with tunable and selective bio- and chemical degradability. AB - A versatile synthetic strategy has been developed which enables the facile incorporation of cleavable functional groups, i.e., esters, thioesters, and disulfides, into the carbon-carbon backbone of vinyl-based polymers. Through the synthesis of novel cyclic monomers, RAFT-mediated radical ring-opening copolymerizations with traditional vinyl monomers such as methyl methacrylate, N,N-dimethylaminoethyl methacrylate, and 2-hydroxyethyl methacrylate lead to the introduction of controlled degradability into these widely used vinyl copolymer systems. An additional benefit of this strategy is the inherent versatility available through the incorporation of cyclic monomers containing diverse functional groups such as esters, thioesters, disulfides, and silyl ether units that allow degradation under basic/acidic, reductive, or enzymatic conditions. By integrating multiple, orthogonal cyclic monomers into linear copolymer backbones, well-defined systems with programmable degradation profiles are obtained which allows for tunable, selective, and stepwise degradation of the vinyl polymer backbones. PMID- 19555104 TI - Synthesis and characterization of a carbon nanotube-dendron series for efficient siRNA delivery. AB - A new series of dendron-functionalized multiwalled carbon nanotube (MWNT) derivatives, characterized by the presence of numerous positively charged tetraalkyl ammonium salts at the periphery of the dendron, has been synthesized. The positive charges on the MWNT surface, coupled with the unique ability of carbon nanotubes (CNTs) to penetrate cell membranes, make the new derivatives potentially ideal vectors for siRNA delivery. Using a fluorescently labeled, noncoding siRNA sequence, we demonstrate that cytoplasmic delivery of the nucleic acid is remarkably increased throughout the different dendron generations. The work reported here highlights the fact that dendron-functionalized CNTs can be rationally designed as efficient carriers of siRNA that can eventually lead to gene silencing. PMID- 19555107 TI - Comparison of aggregation behaviors between ionic liquid-type imidazolium gemini surfactant [C12-4-C12im]Br2 and its monomer [C12mim]Br on silicon wafer. AB - The aggregation of ionic liquid-type imidazolium gemini surfactant [C(12)-4 C(12)im]Br(2) on silicon wafer, which is compared with its monomer [C(12)mim]Br, have been studied. AFM morphology images and contact angle measurements suggest that the aggregations of [C(12)-4-C(12)im]Br(2) and [C(12)mim]Br on silicon wafer follow different mechanisms. Below the critical surface aggregation concentrations (CSAC), both surfactant molecules are adsorbed with their hydrophobic tails facing the air. But above the CSAC, [C(12)-4-C(12)im]Br(2) molecules finally form a bilayer structure with hydrophilic head groups facing the air, whereas [C(12)mim]Br molecules form a multilayer structure, and with increasing its concentration, the layer numbers increase with the hydrophobic chains and hydrophilic head groups facing the air by turns. Besides, the watery wettability of [C(12)-4-C(12)im]Br(2)-treated silica surface is lower than that of [C(12)mim]Br at the concentration of 5.0 cmc, and the infrared spectroscopy suggests that the poorer watery wettability of [C(12)-4-C(12)im]Br(2) may be relative to the less-ordered packing of methylene chains inside the aggregate. These different aggregation behaviors for the two surfactants ascribe to the different molecular structures and electrostatic interactions. This work would have certain theoretical guidance meaning on the modification of solid surface. PMID- 19555109 TI - Studies toward the photochemical synthesis of functionalized [5]- and [6]carbohelicenes. AB - An efficient route to nonsymmetrical helical menthyl esters by means of an oxidative photocyclization reaction of dissymmetric bis-stilbenes is reported. The developed route allows the introduction of functionality on rings A, E, or F, and the influence of the substituent pattern on the photochemical reaction has been examined. Diastereoselectivity is observed when a double chiral induction strategy with dimenthyl helicene esters synthesized in a 70:30 ratio of isomers is used. PMID- 19555106 TI - Transport of free and peptide-bound pyrraline at intestinal and renal epithelial cells. AB - Pyrraline is a quantitatively dominating glycation compound of the advanced Maillard reaction in foods and can be found in urine after consumption of pyrraline-containing food items. The purpose of this study was to investigate the transport of pyrraline and its dipeptide derivatives alanylpyrraline (Ala-Pyrr) and pyrralylalanine (Pyrr-Ala) at intestinal and renal cell lines. Pyrraline inhibited the l-[(3)H]lysine uptake with IC(50) values of 0.3 mM (Caco-2 cells) and 3.5 mM (OK cells), respectively, but not the uptake of [(14)C]Gly-Sar (Caco-2 and SKPT cells). In contrast, Ala-Pyrr strongly inhibited the uptake of [(14)C]Gly-Sar in Caco-2 and SKPT cells with IC(50) values of 0.19 and 0.017 mM, respectively. Pyrr-Ala inhibited the carrier-mediated uptake of [(14)C]Gly-Sar in Caco-2 and SKPT cells by 50% at concentrations of 0.03 and 0.008 mM, respectively. The transepithelial flux of peptide-bound pyrraline across Caco-2 cell monolayers was up to 15-fold higher compared to the flux of free pyrraline. We conclude that free pyrraline is not a substrate for the intestinal lysine transporter and that the absorption of dietary pyrraline occurs most likely in the form of dipeptides rather than as the free amino acid. PMID- 19555105 TI - Dissolution of beech and spruce milled woods in LiCl/DMSO. AB - A novel solvent system, lithium chloride/dimethyl sulfoxide (LiCl/DMSO), was developed for dissolving milled wood. This system completely dissolved beech and spruce milled woods prepared from the Wiley woods (coarse wood meals prepared by a Wiley mill) by 2 h planetary ball-milling under the milling conditions employed. The dependence of the structural change of lignin on the degree of milling was examined to evaluate the correlation between the dissolution and lignin structure. The nitrobenzene oxidation analyses showed that the 2 h milling caused almost no structural change in the aromatic part of lignin in the milled woods. The ozonation analyses suggested that the decrease of the erythro ratio [erythro/(erythro + threo)] obtained from beta-O-4 structure in lignin is only 0.35% after the 2 h milling. Although the yield decrease of the ozonation products was 9.8% after the 2 h milling, this value was fairly smaller than that after a longer time milling. When it is taken into consideration that the other solvent systems for dissolution of wood meal, which are proposed by Lu and Ralph, require 5 h of milling under the same milling conditions to dissolve the milled woods, it is safely stated that the LiCl/DMSO solvent system completely dissolves milled wood, the lignin of which is structurally less altered and, thus, is expected to provide an improved method for structural analysis of the entire lignin fraction in wood cell wall. PMID- 19555110 TI - Temperature dependence of the rate coefficients for the reactions of atomic bromine with toluene, tetrahydrofuran, and tetrahydropyran. AB - The rate coefficients for the reactions of atomic bromine with toluene, tetrahydrofuran, and tetrahydropyran were measured from approximately 295 to 362 K using the relative rate method. Iso-octane was used as the reference compound for the reaction with toluene, and iso-octane and toluene were used as the reference compounds for the reaction with tetrahydrofuran; tetrahydrofuran was used as the reference compound for the reaction with tetrahydropyran. The rate coefficients were found to be unaffected by changes in pressure and oxygen concentration. The rate coefficient ratios were converted to absolute values using the absolute rate coefficient for the reaction of Br with the reference compound. The absolute rate coefficients, in the units cm(3) molecule(-1) s(-1), for the reaction of Br with toluene are given by k(T) = (3.7 +/- 1.7) x 10(-12) exp(-(1.63 +/- 0.15) x 10(3)/T), for the reaction of Br with tetrahydrofuran by k(T) = (3.7 +/- 2.7) x 10(-10) exp(-(2.20 +/- 0.22) x 10(3)/T), and for the reaction of Br with tetrahydropyran by k(T) = (3.6 +/- 1.8) x 10(-10) exp(-(2.35 +/- 0.16) x 10(3)/T). The uncertainties represent one standard deviation. The Arrhenius parameters for these reactions are compared with results in the literature for dimethyl ether, diethyl ether, and a series of saturated hydrocarbons, and the effects of structural variation on these parameters are identified. PMID- 19555111 TI - Localized electrochemistry on a 10 microm spot on a monolith large electrode: an avenue for electrochemical microarray analysis. AB - By measurement of the modulation of ions under an ac field at the electrode/electrolyte interface layer (EIL) at a spatial modulation of 0.0001 nm, a quantitative measurement of local redox current on a 10 microm spot of a working electrode is achieved at a sensitivity of 0.02 nA/spot. The spot is defined by the probe laser beam. We show the principle for two redox ions, [Fe(CN)(6)](3-) and [Ru(NH(3))(6)](3+), on a 4 mm(2) Au electrode. The redox peaks from the optical and the electrochemical (cyclic voltammetry (CV)) responses, which are concomitantly measured, are within 0.005 V. The setup is identical to the conventional three-electrode measurement, and the laser beam does not perturb the electrochemical process. We demonstrate high sensitivity to "read" binding between two single-stranded DNA (ssDNA) on a 10 microm spot for DNA microarray analysis where the signal is "blind" to nonspecific binding. Potentially, quantitative combinatorial electrochemistry can be performed on a monolith electrode by scanning the laser beam on the electrode. PMID- 19555112 TI - Optical Zeeman spectroscopy of ytterbium monofluoride, YbF. AB - The Zeeman-induced shifts and splittings of low-J lines in the (O)P(12) branch of the (0,0) band of the A(2)Pi(1/2)-X(2)Sigma(+) electronic transition of a cold molecular beam sample of ytterbium monofluoride, YbF, have been recorded. The Zeeman spectra for the (171)YbF, (172)YbF, and (174)YbF isotopologues have been analyzed using a standard effective Hamiltonian approach. The magnetic g-factors determined for the A(2)Pi(1/2)(v = 0) state are rationalized using the predicted and observed electronic state distribution. The observed Zeeman tuning of the levels in the A(2)Pi(1/2)(v = 0) state is unexpectedly large; this is caused by mixing with the B(2)Sigma(+) state. PMID- 19555113 TI - Cell division cycle 7 kinase inhibitors: 1H-pyrrolo[2,3-b]pyridines, synthesis and structure-activity relationships. AB - Cdc7 kinase has recently emerged as an attractive target for cancer therapy and low-molecular-weight inhibitors of Cdc7 kinase have been found to be effective in the inhibition of tumor growth in animal models. In this paper, we describe synthesis and structure-activity relationships of new 1H-pyrrolo[2,3-b]pyridine derivatives identified as inhibitors of Cdc7 kinase. Progress from (Z)-2-phenyl-5 (1H-pyrrolo[2,3-b]pyridin-3-ylmethylene)-3,5-dihydro-4H-imidazol-4-one (1) to [(Z)-2-(benzylamino)-5-(1H-pyrrolo[2,3-b]pyridin-3-ylmethylene)-1,3-thiazol-4(5H) one] (42), a potent ATP mimetic inhibitor of Cdc7 kinase with IC(50) value of 7 nM, is also reported. PMID- 19555114 TI - Synthesis, properties, and mechanistic insight into the self-assembly of a lamellar fibrous superstructure from a synthetically simple discotic molecule. AB - The discotic molecule 4-chloro-2,6-bis(octadecylamino)-pyrimidine-5-carbaldehyde, displays gelation behavior in dodecane, heptane, chloroform, and dichloromethane. The aggregation behavior of this material was studied by dynamic light scattering, differential scanning calorimetry, scanning electron microscopy, polarized optical microscopy, small-angle X-ray diffraction, and wide-angle X-ray diffraction techniques. Combined with molecular modeling calculations, Fourier transform infrared, and 1H NMR studies, we propose a mechanism for the self assembly of this fibrous lamellar architecture. Notably, we have shown that the fibers grow via stacking interactions along their main axis, via hydrogen bonding along their short axis, and via van der Waals interactions (lamellae) along the third axis. This type of morphology is desirable since it provides an opportunity to synthetically control and optimize mechanical, electrical, optical, and transport properties along the length of the fiber. PMID- 19555115 TI - Total synthesis of (-)-himandrine. AB - We describe the first total synthesis of (-)-himandrine, a member of the class II galbulimima alkaloids. Noteworthy features of this chemistry include a diastereoselective Diels-Alder reaction in the rapid synthesis of the tricycle ABC-ring system in an enantiomerically enriched form, the use of a formal [3+3] annulation strategy to secure the CDE-ring system with complete diastereoselection, and successful implementation of our biogenetically inspired oxidative spirocyclization of an advanced intermediate. The successful and direct late-stage formation of the F-ring in the hexacyclic core of himandrine drew on the power of biogenetic considerations and fully utilized the inherent chemistry of a plausible biosynthetic intermediate. PMID- 19555116 TI - New strategy for the screening of lysosomal storage disorders: the use of the online trapping-and-cleanup liquid chromatography/mass spectrometry. AB - The aim of this study was to set up a robust method suitable for large-scale studies (screening) with a minimized preparation process and with reduced running costs, for measuring five enzyme activities on dried blood spots by a new and simplified tandem mass spectrometry-based method. After incubation, all 5 reaction mixtures, carried out separately, were stopped, combined together, and centrifuged. The cleaning-up of the injected mixture was performed through a fast online trapping step preceding a liquid chromatography/tandem mass-spectrometry measurement. This method takes only 4 min as analysis run time and without any purification following the enzymatic reaction. We assessed the effectiveness of this approach in assaying the enzymatic activities on dried blood spots from 10 patients affected by "Pompe", 6 by "Gaucher", 12 by "Fabry", 3 by "Niemann-Pick" A/B, and 2 by "Krabbe" diseases. Reference values were established on 5000 healthy newborns and 300 healthy adults. All affected patients showed enzymatic activities below the normal range. In heterozygous carriers (18 for Fabry, 10 for Pompe, and 4 for Gaucher disease) the activities were slightly lower than in control subjects. The results show that the method set out in its simplicity, low costs, and low processes preparations can be fully applicable to a mass screening. PMID- 19555117 TI - A versatile route to the tulearin class of macrolactones: synthesis of a stereoisomer of tulearin A. AB - A versatile synthetic approach to the tulearin class of macrolactones has been developed and deployed to make a stereoisomer of tulearin A. The knowledge gained about structure and synthesis will expedite the assignment of the stereostructure of this new anticancer agent. PMID- 19555119 TI - Crystal structure of an archaeal Rad51 homologue in complex with a metatungstate inhibitor. AB - Archaeal RadAs are close homologues of eukaryal Rad51s ( approximately 40% sequence identities). These recombinases promote a hallmark strand exchange process between homologous single-stranded and double-stranded DNA substrates. This DNA-repairing function also plays a key role in cancer cells' resistance to chemo- and radiotherapy. Inhibition of the strand exchange process may render cancer cells more susceptible to therapeutic treatment. We found that metatungstate is a potent inhibitor of RadA from Methanococcus voltae. The tungsten cluster binds RadA in the axial DNA-binding groove. This polyanionic species appears to inhibit RadA by locking the protein in its inactive conformation. PMID- 19555118 TI - Arrestin2/clathrin interaction is regulated by key N- and C-terminal regions in arrestin2. AB - The interaction of nonvisual arrestins with clathrin is an important step in mediating the endocytosis of cell surface receptors. Previous studies have shown that mutation of the clathrin-binding box in arrestin leads to severe defects in arrestin-mediated trafficking. However, little is known about how arrestin/clathrin interaction is regulated. Here we show that both the N- and C terminal regions of arrestin2 function to inhibit basal interaction with clathrin. Truncation analysis revealed that clathrin binding increases as the C tail of arrestin2 is shortened while site-directed mutagenesis identified Glu 404, Glu-405, and Glu-406 as being primarily responsible for this inhibition. Mutagenesis also identified Lys-4, Arg-7, Lys-10, and Lys-11 within the N terminus as playing a key role in regulating clathrin binding. Based on similarities with visual arrestin, Lys-10 and Lys-11 likely function as phospho sensors in arrestin2 to initially discriminate the phosphorylation status of target receptors. Analysis of the arrestin2 structure reveals that Arg-7, Lys-10, and Lys-11 are in close proximity to Glu-389 and Asp-390, suggesting that these residues may form intramolecular interactions. In fact, simultaneous mutation of Glu-389 and Asp-390 also leads to enhanced clathrin binding. These results reveal that multiple intramolecular interactions coordinately regulate arrestin2 interaction with clathrin, highlighting this interaction as a critical step in regulating receptor trafficking. PMID- 19555121 TI - Scuteflorins A and B, dihydropyranocoumarins from Scutellaria lateriflora. AB - Two new dihydropyranocoumarins, scuteflorins A (1) and B (2), together with the known compounds decursin (3), chrysin (4), oroxylin A (5), wogonin (6), 5,7 dihydroxy-8,2'-dimethoxyflavone, dihydrochrysin, dihydrooroxylin A, lupenol, scutellaric acid, pomolic acid, ursolic acid, beta-sitosterol, daucosterol, and palmitic acid, were isolated from the aerial parts of Scutellaria lateriflora, commonly used as a dietary supplement. The structures of 1 and 2 were established by means of 1D and 2D NMR spectra as well as HRMS data. The absolute configuration of coumarins 1 and 2 was determined by comparison of experimental and theoretical calculated CD spectra. The cytotoxicity and antioxidant effects of the methanol extract of this plant and some of the constituent flavonoids were evaluated in vitro. PMID- 19555120 TI - Botryllamides: natural product inhibitors of ABCG2. AB - ABCG2 is a membrane-localized, human transporter protein that has been demonstrated to reduce the intracellular accumulation of substrates through ATP dependent efflux. Highly expressed in placental syncytiotrophoblasts, brain microvasculature, and the gastrointestinal tract, ABCG2 has been shown to mediate normal tissue protection as well as limit oral bioavailability of substrate compounds. Development of ABCG2 inhibitors for clinical use may allow increased penetration of therapeutic agents into sanctuary sites and increased gastrointestinal absorption. Previously identified inhibitors have lacked potency or specificity or were toxic at concentrations needed to inhibit ABCG2; none are in clinical development. A previously developed high-throughput assay measuring inhibition of ABCG2-mediated pheophorbide a transport was applied to natural product extract libraries. Among the active samples were extracts from the marine ascidian Botryllus tyreus. Bioassay-guided fractionation resulted in purification of a series of botryllamides. Ten botryllamides were obtained, two of which (designated I and J) were novel. Activity against ABCG2 was confirmed by assessing the ability of the compounds to inhibit ABCG2-mediated BODIPY-prazosin transport in ABCG2-transfected HEK293 cells, compete with [(125)I] iodoarylazidoprazosin (IAAP) labeling of ABCG2, stimulate ABCG2-associated ATPase activity, and reverse ABCG2-mediated resistance. PMID- 19555122 TI - Cardenolides from Pergularia tomentosa display cytotoxic activity resulting from their potent inhibition of Na+/K+-ATPase. AB - Two new cardenolide glycosides (1 and 2), along with six known cardenolide glycosides (3-8), have been isolated from the roots of Pergularia tomentosa. In order to investigate their potential anticancer activity, these compounds were tested in an in vitro growth inhibitory assay (a MTT colorimetric assay), including six different human cancer cell lines, and for their ability to inhibit Na(+)/K(+)-ATPase activity, in addition to the morphologic changes induced in human cancer cell lines (using computer-assisted phase-contrast microscopy). The data revealed that these cardenolides displayed marked cytotoxic activity. The results obtained suggest that structural characteristics of the cardenolides studied, with the A/B rings of the steroidal skeleton trans fused and containing a single sugar in a unique "dioxanoid" attachment, confer on them specific cytotoxic properties that are distinct from those displayed by classic cardenolides such as digoxin. PMID- 19555123 TI - Cytotoxic and antimycobacterial prenylated flavonoids from the roots of Eriosema chinense. AB - Eight new prenylated flavonoids, khonklonginols A-H (1-8), together with six known compounds including five flavonoids, lupinifolinol (9), dehydrolupinifolinol (10), flemichin D (11), eriosemaone A (12), and lupinifolin (13), and one lignan, yangambin (14), have been isolated from hexane and dichloromethane extracts of the roots of Eriosema chinense. The structures of 1-8 were elucidated by spectroscopic methods. The compounds were evaluated for cytotoxic activity against the small-cell lung (NCI-H187) and oral epidermal carcinoma (KB) human cell lines as well as for antimycobacterial activity against Mycobacterium tuberculosis H37Ra. PMID- 19555124 TI - Labdane diterpenoid glycosides from Alpinia densespicata and their nitric oxide inhibitory activities in macrophages. AB - Four new labdane diterpene glycosides (1-4) named alpindenosides A, B, C, and D, two new norditerpene glycosides (5, 6) named noralpindenosides A and B, and three known flavonoid glycosides (7-9) have been isolated from the stems of Alpinia densespicata. Structural elucidation of compounds 1-6 was based on spectroscopic data. Compounds 1-9 all exhibited moderate NO inhibitory activities, whereas they were noncytotoxic at 20 microM against several human tumor cell lines. PMID- 19555125 TI - Tyrosinase-inhibitory constituents from the twigs of Cinnamomum cassia. AB - A methanol extract of the twigs of Cinnamomum cassia was found to possess inhibitory activity against tyrosinase. Purification of the MeOH extract afforded four new phenolics, cassiferaldehyde (6), icariside DC (9), cinnacassinol (10), and dihydrocinnacasside (13), together with 10 known compounds (1-5, 7-12, and 14). The structures of the new compounds were determined by spectroscopic data interpretation. Compounds 1-6 and 8-13 showed strong inhibitory activity against tyrosinase, with IC(50) values ranging from 0.24 to 0.94 mM. PMID- 19555129 TI - Childhood obesity and elevated blood pressure in a rural population of northern Greece. AB - INTRODUCTION: The objective of this study was to determine the prevalence of childhood obesity and elevated blood pressure (BP) in a rural population of northern Greece. METHODS: In total, 572 schoolchildren between the age of 4 and 10 years were examined. Obesity was defined using three different standards: (1) body mass index (BMI) charts of the French society of Paediatrics (FR), selected because of the low cardiovascular risk profile and low prevalence of obesity in France; (2) United States BMI CDC charts (US), selected because of the high prevalence of childhood obesity in the USA; and the reference curves of the International Obesity Task Force (IOTF). Children with elevated BP were defined as BP > or = 95th percentile for age, gender and height, according to the Greek national charts. RESULTS: The prevalence of obesity for boys was 13.6% (IOTF), 23.7% (US) and 31.7% (FR); for girls 14.4% (IOTF), 21.1% (US) and 35.1% (FR). The prevalence of elevated BP was 7.9% (45 children). It was 5 to 6 times more common for obese than non-obese children to have elevated BP (relative risk of 5.2 to 6.2 and odds ratio 6.3 to 7.7). CONCLUSIONS: The results confirm the high prevalence of childhood obesity in Greece, in this study found to be more prevalent in rural than urban Greece. The IOTF criteria tend to underestimate obesity and may not be optimal for use in a primary clinical care setting where the approach is for health education and patient treatment, rather than purely epidemiological. The study also confirms a strong relationship between high BP and increased BMI. PMID- 19555126 TI - Apogossypol derivatives as pan-active inhibitors of antiapoptotic B-cell lymphoma/leukemia-2 (Bcl-2) family proteins. AB - Guided by nuclear magnetic resonance (NMR) binding assays and computational docking studies, a series of 5,5' substituted apogossypol derivatives was synthesized that resulted in potent pan-active inhibitors of antiapoptotic Bcl-2 family proteins. Compound 8r inhibits the binding of BH3 peptides to Bcl-X(L), Bcl-2, Mcl-1, and Bfl-1 with IC(50) values of 0.76, 0.32, 0.28, and 0.73 microM, respectively. The compound also potently inhibits cell growth of human lung cancer and BP3 human B-cell lymphoma cell lines with EC(50) values of 0.33 and 0.66 microM, respectively. Compound 8r shows little cytotoxicity against bax(-/ )bak(-/-) cells, indicating that it kills cancers cells via the intended mechanism. The compound also displays in vivo efficacy in transgenic mice in which Bcl-2 is overexpressed in splenic B-cells. Together with its improved chemical, plasma, and microsomal stability relative to compound 2 (apogossypol), compound 8r represents a promising drug lead for the development of novel apoptosis-based therapies for cancer. PMID- 19555130 TI - Recognizing the rising impact of diabetes in seniors and implications for its management. AB - OBJECTIVES: To identify the incidence and symptoms of type 2 diabetes in older adult patients, review appropriate glycemic targets for older adults with type 2 diabetes, and consider the rationale for managing hyperglycemia in this patient population. DATA SOURCES: Live symposium presentation based on clinical practice and research, medical literature, and studies published between January 1993 and November 2008 on managing diabetes in older adults, government statistics, and medical society guidelines. STUDY SELECTION: Thirty-five articles were identified from various data sources and were evaluated. The following search terms were used: Complications, Diabetes, Glucose Tolerance, Goals, Hyperglycemic Hyperosmolar Nonketotic Syndrome (HHNS), Long-term Care, Management, Obesity, Prevalence, Risk Factors, Screening, Senior, and Symptoms. All information deemed relevant to recognizing the rising impact of diabetes in seniors and its implications for management were included. DATA EXTRACTION: Data were extracted independently of the author by the librarians of the University of Pittsburgh. DATA SYNTHESIS: As a result of the aging of the U.S. population, the increasing prevalence of diabetes, and the considerable impact of this disease among older adults, it is only logical to assume that the number of older individuals with diabetes and its impact on older adults will continue to rise. Uncontrolled diabetes has significant risks in the elderly beyond the typical microvascular and macrovascular complications commonly associated with diabetes: cognitive impairment, depression, excessive skin problems, and an increased risk of falls. However, glycemic control reduces the development of these complications and can lead to improvements in dementia, memory, energy, physical activity, mood, and quality of life. CONCLUSION: Because many older adults may benefit from intensive, long-term glycemic control, consultant pharmacists should understand the importance of individualizing glycemic targets, management strategies, and pharmacotherapy in older adults. PMID- 19555131 TI - When oral agents fail: optimizing insulin therapy in the older adult. AB - OBJECTIVES: To evaluate strategies for managing older adults with type 2 diabetes who are inadequately controlled on oral agents and to consider when and how to transition older adults from usual care with oral agents to insulin therapy or incretin-based therapy. DATA SOURCES: Live symposium presentation based on clinical practice and research, medical literature, and studies published between November 1999 and November 2008 on managing diabetes in older adults, government statistics, and medical society guidelines. STUDY SELECTION: A study selection was performed using the following search terms: Basal Insulin, Diabetes, Elderly, Incretins, Insulin, Insulin Pharmacokinetics, Long-Term Care, Long-Term Complications, Medications, Oral antidiabetic, Senior, Treatment Goals. Articles were identified from various data sources containing information relevant to the early initiation of insulin and incretin-based drugs in the elderly as well as goals and treatment options in this group. DATA EXTRACTION: Data were extracted by the author and Nicole Cooper (DesignWrite, LLC). DATA SYNTHESIS: Recent guidelines suggest the need for early initiation of insulin in patients who fail to achieve or maintain goals on one or more oral agents. Basal-insulin therapy is convenient and appropriate for many older individuals who fail one or more oral agents. Prandial insulin coverage with basal-bolus insulin therapy or premixed insulin therapy may be useful in patients with significant postprandial hyperglycemia. Patients who need postprandial control also may benefit from using a glucagon-like peptide-1 (GLP-1) analog or a dipeptidyl peptidase-4 (DPP-4) inhibitor because they are unlikely to cause hypoglycemia, but reductions in hyperglycemia with these agents are limited. CONCLUSION: Basal insulin therapy, basal-bolus insulin therapy, and premixed insulin therapy are useful options for older patients inadequately controlled on one or more oral agents. Older patients who require additional postprandial control may benefit from GLP-1 analogs or DPP 4 inhibitors. Additional research is needed to determine optimal therapy in older adults treated with oral agents who require more intensive therapy. PMID- 19555132 TI - Minimizing the risk of hypoglycemia in older adults: a focus on long-term care. AB - OBJECTIVES: To consider the risk and impact of hypoglycemia in older adults with type 2 diabetes (T2DM), to describe how to interpret blood glucose readings in an older patient's medical record, and to discuss strategies for avoiding hypoglycemia in this patient population. DATA SOURCES: Live symposium presentation based on clinical practice and research, medical literature, and studies published between January 1990 and November 2008 on managing T2DM in older adults, government statistics, and medical society guidelines. STUDY SELECTION: A literature search was performed. Search terms included: Diabetes, Elderly, Hypoglycemia, Insulin Pattern Management, Long-Term Care, Senior, Sliding Scale Insulin. Twenty-two articles were identified from various sources containing information relevant to the identification and treatment of hypoglycemia in elderly persons with diabetes. DATA EXTRACTION: Data were extracted by the author and by Nicole Cooper (DesignWrite, LLC). DATA SYNTHESIS: Numerous studies have demonstrated the benefits of achieving near-normal levels of glycemic control in patients with T2DM. However, avoiding hypoglycemia during intensive therapy can be challenging for older adults and may present a barrier to glycemic control in this population. Pharmacists working in long-term care settings can take several steps to help their older patients avoid hypoglycemia, including recommending the use of insulin regimens with more physiologic time action profiles, such as insulin analogs. They also can help facilities develop protocols for treating hyperglycemia and hypoglycemia, offer insulin pattern management services, and help educate the staff about the optimal use of insulin therapies. CONCLUSIONS: Consultant pharmacists can play a critical role in preventing hypoglycemia in long-term care facilities by recommending the use of more physiologic insulin regimens, developing facility protocols for glycemic management, and providing staff education. PMID- 19555133 TI - Rethinking code blue in long-term care. AB - Losing a loved one is often emotional and painful for families, and most aspects of death and dying are usually difficult for them to discuss. Our traditional view of death (as a failure) is being reassessed. Many residents' conditions place them at high risk for death, or they may have conditions considered terminal. Numerous facilities are rethinking their approach to Code Blue, and this is an ideal time to analyze the entire process, especially since death is a frequent occurrence in long-term care facilities. Approximately 10% of residents admitted under the Medicare benefit die, or are hospitalized and subsequently die, within 30 days of admission. In addition to simplifying rescue techniques, a movement is afoot to allow family members into scenes previously considered sacrosanct by medical care providers. PMID- 19555134 TI - Lights, camera, promotion: making media relations work. AB - Media relations isn't part of the typical pharmacy school curriculum, but communicating with the media is a skill that senior care pharmacists need to get their message out to consumers. These practitioners must be able to conduct effective interviews, develop relationships with reporters and producers, and establish themselves as experts in their fields. When pharmacists are media savvy, they enjoy the benefit of increased visibility and opportunities to positively influence safe medication use by consumers nationwide. PMID- 19555135 TI - Incidence of abnormal metabolic parameters and weight gain induced by atypical antipsychotics in elderly patients with dementia. AB - OBJECTIVE: To determine if atypical antipsychotic therapy leads to the development of abnormal metabolic parameters and weight gain in elderly patients with dementia. DESIGN: Retrospective chart review. SETTING: Veterans Affairs Medical Center. PATIENTS: Veterans 65 years of age or older with the diagnosis of dementia. MAIN OUTCOME MEASURE: The incidence of impaired fasting glucose (> 100 mg/dL) after initiation of atypical antipsychotic therapy. The secondary objectives were to determine the incidence of significant weight gain, worsening of lipid values, new onset of type 2 diabetes mellitus, and metabolic syndrome. RESULTS: After reviewing 979 charts for inclusion and exclusion criteria, 56 patients were found eligible for the study. More than 50% of patients were excluded because they were lacking baseline or follow-up glucose laboratory results. Ten percent of the study population developed impaired fasting glucose after starting atypical antipsychotic therapy. Overall glucose increased by 9.7 mg/dL from baseline to follow-up. Significant weight gain (>or= 7% of baseline weight) occurred in 8.92% of elderly. However, overall weight decreased by 1.3 kg during the study periods. Patients who developed worsening lipid parameters or were started on lipid-lowering therapy were 14.5% of the study population even though overall lipid levels improved or remained unchanged. CONCLUSION: Periodic monitoring of glucose should be considered for patients with dementia begun on atypical antipsychotics, although aggressive monitoring may be controversial for end-stage dementia. Overall, weight reduction and improvement in lipid parameters were observed in this study. The common metabolic adverse effects noted frequently with atypical antipsychotics may not be as much of a concern with the elderly population. PMID- 19555136 TI - Methylnaltrexone methobromide: the first peripherally active, centrally inactive opioid receptor-antagonist. AB - OBJECTIVE: To review the chemistry, pharmacodynamics, pharmacokinetics, clinical efficacy, tolerability, indications, and dosing and administration of methylnaltrexone methobromide, the first approved peripherally selective opioid receptor-antagonist. DATA SOURCE: MEDLINE/PUBMED and EMBASE searches (January 1966-February 2009) were conducted to identify pertinent English-language studies. STUDY SELECTION AND DATA EXTRACTION: All studies evaluating any aspect of methylnaltrexone methobromide. DATA SYNTHESIS: Subcutaneous methylnaltrexone methobromide is the first opioid receptor-antagonist to be approved for the treatment of opioid receptor-agonist-induced constipation (subset with advanced disease, receiving palliative care, with an inadequate response to laxative therapy). This agent lacks meaningful activity in the central nervous system and, hence, will not compromise centrally mediated analgesia or precipitate centrally mediated signs/symptoms of opioid receptor-agonist withdrawal. There are no published comparative trials with traditional pharmacologic/nonpharmacologic laxation regimens. CONCLUSION: Methylnaltrexone methobromide is administered into the upper arm, abdomen, or thigh once every other day, with the frequency of dosing being increased, if needed, to a maximum of once daily. Recommended doses are 8 mg, 12 mg, or 0.15 mg/kg, depending on patient weight. For creatinine clearances less than 30 mL/min, the dose should be reduced by 50%. The average wholesale price is $83.33 for a 12 mg single-use vial (Medispan, accessed December 4, 2008). Clearly, parenteral agents are not as useful as oral agents and results of ongoing studies with new oral formulations of this product are eagerly awaited. PMID- 19555137 TI - Warfarin potentiation: a review of the "FAB-4" significant drug interactions. AB - Warfarin is an anticoagulant with numerous drug-drug interactions. Four significant drug interactions with warfarin are: fluconazole, amiodarone, sulfamethoxazole/trimethoprim, and metronidazole. Potentially life-threatening interactions with these medications are a common occurrence because they have a broad spectrum of indications in elderly individuals. With the increasing number of elderly individuals, who consume a disproportionate number of medications, cautious monitoring for these drug interactions is essential. Drug interactions often are overlooked despite computer alert systems. Careful analysis of each patient's situation will help in evaluating whether the use of a medication is appropriate. Through knowledge of potential mechanisms of interaction, management options such as dose reductions and/or the use of alternative agents should be used in determining the appropriate drug therapy. PMID- 19555138 TI - Preventing and treating VTE in the elderly patient. AB - While deaths from cardiovascular disease have decreased dramatically over the past several decades, mortality and morbidity from venous thromboembolism (VTE) continues to threaten hundreds of thousands of patients in the United States each year. Because the risk of VTE increases with age, the problem will only worsen as the population ages. Raising the awareness of health care practitioners and patients about the ways to prevent and treat VTE-including the most recent changes to the guidelines issued by the American College of Chest Physicians-is critical to successfully addressing this significant public health problem. PMID- 19555139 TI - New drug update: 2008. AB - Only 16 new therapeutic agents were marketed in the United States in 2008, the lowest number in many years. Four new drugs have been selected for consideration in this review. The uses and most important properties of each agent are considered, and a rating for each new drug is provided using the New Drug Comparison Rating (NDCR) system developed by the author. PMID- 19555140 TI - Screening for undiagnosed cognitive impairment in homebound older adults. AB - OBJECTIVE: To screen for undetected cognitive impairment in homebound elders receiving home health care services. DESIGN: A cross-sectional study of adults 60 years of age and older receiving visiting nurse services. A pharmacist administered the Mini-Cog, a rapid screening test for cognitive impairment, during the enrollment home visit. SETTING: Participants' homes. PARTICIPANTS: A consecutive sample of 100 homebound adults, 60 years of age or older with no previous diagnosis of dementia, Alzheimer's disease, or other cognitive impairment. MAIN OUTCOME MEASURE: Percentage of participants scoring in the likely impaired range (screen failure) on the Mini-Cog. RESULTS: Seventeen percent of persons with no prior diagnosis of dementia or cognitive decline failed the Mini-Cog. CONCLUSION: Rates of undetected cognitive impairment are substantial in homebound elders receiving care from a visiting nurse service. The home health setting represents an important point in the continuum of geriatric care for detection of cognitive impairment. Future work should define the types and trajectories of cognitive impairment detected in home care patients by simple screens such as the Mini-Cog and test ways to integrate this knowledge into longitudinal treatment plans across settings of care. PMID- 19555141 TI - Identifying, preventing, and reporting elder abuse. AB - Elder abuse and neglect is a national public health problem with a profound negative impact on community health. Neglect is its most common manifestation, but other types of overt abuse are frequent. National initiatives are underway to use trained professionals, physicians and others, to act as community sentinels to uncover and help unknown victims. Pharmacy, with high visibility and easy access to seniors, must join other professions in this initiative to protect the vulnerable elderly. Reporting suspected abuse-the major tool of prevention-is a simple and time-efficient, though an often misunderstood, process. This article will teach community and consultant pharmacists to identify, prevent, and report senior abuse and discuss how patients benefit from such action. Abuse in institutionalized settings, which has wide coverage in the literature, will not be reviewed here. PMID- 19555142 TI - Recognizing Charles Bonnet syndrome in the elderly. AB - Charles Bonnet syndrome (CBS) is an under-recognized and commonly misdiagnosed condition characterized by the presence of visual hallucinations that psychologically normal people acknowledge as being unreal. It is commonly associated with ocular pathology and usually observed in elderly individuals with visual impairment. The exact etiology of CBS is unknown; however, the presentation of hallucinations is believed to be a result of functional deterioration of the central and peripheral nervous systems. Eradication of hallucinations and recurrent episodes has been seen with the use of neuroleptic and anticonvulsant agents. Correction of underlying ocular disorders and low vision rehabilitation may also help in the resolution of visions. Careful patient assessment is necessary to appropriately diagnose CBS and determine the best approach to management. PMID- 19555143 TI - Kidney stones: painful and common--but preventable. AB - Kidney stones are among the more common and painful urological disorders, affecting approximately 10% of Americans. While most stones pass spontaneously, obstructions result in renal colic, often requiring aggressive management of pain. If obstructions persist, extracorporeal shockwave lithotripsy, ureteroscopy, or percutaneous nephrolithotomy may be required for a stone's removal. Recurrence rates are high, and prevention includes increased fluid intake, dietary restrictions, and the use of pharmacotherapy. Stone recurrence increases with age, underscoring the importance of prevention for those 60 years of age and older. PMID- 19555144 TI - Drug-abuse deterrent formulations. AB - Prescription drug abuse is a large and growing problem. In the decade between 1992 and 2002, prescriptions for controlled substances increased roughly three times faster than prescriptions for noncontrolled substances and 12 times faster than the general population. With drug/formulation tampering now a prevalent problem, regulators and manufacturers have turned to the manufacturing process for ways to decrease the likelihood that a specific formulation is abuse-ready. Abuse-deterrent formulations are one aspect of a comprehensive approach to prescription drug risk management. Several approaches seem most likely to deter misuse and abuse: adding agonists or excipients that foil tampering efforts, using physical barriers, and designing prodrugs. PMID- 19555145 TI - Mark H. Beers: an appreciation. AB - Mark H. Beers, MD, revolutionized the way medications are prescribed for the elderly. The list that bears his name, the "Beers criteria," became the watchword for practitioners, particularly consultant pharmacists, who oversee drug therapy for the elderly in nursing facilities and in outpatient settings. An honorary lifetime member of the American Society of Consultant Pharmacists, his cross disciplinary approach was crucial to the success of his efforts. PMID- 19555146 TI - Concurrent use of statins and amiodarone. AB - OBJECTIVE: To estimate the concurrent use between statins and amiodarone in context with published case reports of drug-interaction-induced rhabdomyolysis. DESIGN: Retrospective analysis of a longitudinal prescription claims database for concurrent prescriptions of statins and amiodarone dispensed during 2006. PATIENTS, PARTICIPANTS: The study population includes an unprojected annual number of patients who filled a prescription for an HMG CoA reductase inhibitor or simvastatin-containing products or lovastatin-containing products or Lipitor (atorvastatin) or Caduet (amlodipine/atorvastatin) concurrently with brand and generic forms of amiodarone during 2006. The concurrency analysis was used to provide context for published case reports of rhabdomyolysis/myopathy related to simvastatin and amiodarone concurrent use. MAIN OUTCOME MEASURE: Episodes of concurrent use between statins and amiodarone. RESULTS: Findings from this analysis indicate noteworthy amiodarone and statin concurrency (44%) when based on amiodarone patient volume. Atorvastatin had the greatest level of concurrency (23.5%) with amiodarone followed by simvastatin (13.3%). Proportionality based on amiodarone patient volume shows a greater level of concurrency with 20 mg (6%) and 40 mg (5.5%) simvastatin strengths compared with other simvastatin strengths. CONCLUSION: Clinicians should be vigilant in monitoring the regimens of patients prescribed a statin with drugs that may increase the risk of myopathy. In particular, since nearly half of the patients prescribed amiodarone may also be prescribed a statin, then addition of amiodarone or changes in statin dose should trigger a drug regimen review and patient level monitoring. Clinicians should avoid simvastatin doses greater than 20 mg per day in patients taking amiodarone. PMID- 19555148 TI - Cardiovascular risk and TZD: safe therapy for the elderly? AB - Recent data regarding cardiovascular risks have raised serious safety concerns with thiazolidinedione (rosiglitazone and pioglitazone) therapy. Some studies have identified an increased risk of myocardial infarction and death with rosiglitazone use; others found no increased risk. Multiple comorbidities (heart failure, renal insufficiency) limit diabetes treatment options in the elderly. It is estimated that more than 30% of nursing facility residents have diabetes; therefore, pharmacists can benefit from a review of safety concerns with thiazolidinediones. Research findings reported by the media can be misinterpreted by laypersons, making pharmacists an integral resource in answering concerns about thiazolidinedione safety. PMID- 19555147 TI - Vaccines for older adults. AB - OBJECTIVE: To update senior care pharmacists with current recommendations on influenza, pneumococcal, zoster, and tetanus vaccines for older adults 65 years of age or older. DATA SOURCES: A search of immunization guidelines on influenza, pneumococcal, zoster, and tetanus infection was performed at various Web sites, including Immunization Action Coalition (www.immunize.org) and the Centers for Disease Control and Prevention (www.cdc.gov). MEDLINE (January 1966 to September 2008) and Pub-Med (January 1955-September 2008) databases and manual bibliographic searches were used to identify additional information. Search terms included influenza, herpes zoster, shingles, pneumococcal pneumonia, pneumovax, tetanus, and vaccine, used alone or in combination. STUDY SELECTION AND DATA EXTRACTION: Relevant English language articles identified from the data sources were evaluated. DATA SYNTHESIS: Trivalent influenza vaccine is recommended to high-risk groups and persons who are likely to transmit the infection, including selected health care staff and caretakers, adults 50 years of age or older, and those with comorbidities that increase the risk of influenza complications. Pneumococcal vaccine is recommended for all persons 65 years of age or older and those with certain medical conditions. One-time revaccination can be offered to the elderly who initially received the vaccine before they turned age 65 and at least five years passed since the initial vaccination. Zoster vaccine should be offered to all persons 60 years of age or older unless contraindications or precautions exist. Tetanus booster every 10 years should be considered in adults who have completed the primary series. Influenza and pneumococcal vaccines, as well as tetanus booster for post-exposure management, are covered under Medicare Part B. Zoster vaccine is covered under Medicare Part D. CONCLUSION: The Advisory Committee on Immunization Practices offers detailed recommendations for the prevention of influenza, invasive pneumococcal disease, zoster, and tetanus infection for target groups. PMID- 19555149 TI - Defining and diagnosing epilepsy in the elderly. AB - OBJECTIVES: To distinguish between nonepileptic and epileptic seizures and further categorize epileptic seizures. To describe the prevalence and risk factors related to epilepsy in the elderly, including seizure triggers. To explain how to evaluate elderly patients who present with seizure symptoms. DATA SOURCES: Live symposium presentation based on clinical practice and research. Current clinical research and guidelines. CONCLUSIONS: Seizures and epilepsy are prevalent in the elderly (> or =65 years of age). It is important to recognize that seizures in the elderly may present differently and can mimic other conditions common in the aged. Consultant pharmacists should understand these differences and be able to classify seizure types to allow for more appropriate treatment and management recommendations. PMID- 19555150 TI - Epilepsy in the elderly: medications and pharmacokinetics. AB - OBJECTIVES: To describe the pharmacokinetics of the different antiepileptic drugs. To discuss optimal pharmacologic use of antiepileptic therapy as it relates to elderly individuals taking multiple medications. To understand the controversy over generic versus brand-name antiepileptic drugs. DATA SOURCES: Live symposium presentation based on clinical practice and research. CONCLUSIONS: While the goal of antiepileptic drug therapy is to have patients be seizure-free without experiencing side effects, this ideal may be difficult to achieve. Classical and newer antiepileptic drugs have varying mechanisms of action and pharmacokinetics that consultant pharmacists must understand. Because of the potential of drug interactions, selecting an appropriate antiepileptic drug requires a thorough assessment of patient health history to determine which therapy will be most effective and safe. Generic options are available and are effective for some, but concerns about bioequivalence make their use controversial. PMID- 19555151 TI - Managing epilepsy: issues in the elderly. AB - OBJECTIVES: To describe key factors that can affect treatment of epilepsy in the elderly. To outline steps to reduce or discontinue antiepileptic drug therapy in those who have been seizure-free. To design a care plan for a resident with a diagnosis of epilepsy. DATA SOURCES: Live symposium presentation based on clinical practice and research. CONCLUSIONS: Treating and managing epilepsy in the elderly may be complicated by confounding factors, including comorbidities and polypharmacy. Special attention must be paid to reduce drug interactions and monitor for effectiveness and toxicity. Consultant pharmacists play a key role in managing patients with epilepsy, and nursing facilities should strive to integrate consultant pharmacists into all stages of care plan development and revision. PMID- 19555152 TI - The consultant pharmacist's expanded role in nutrition management. AB - Recent changes to the State Operations Manual have expanded the role of consultant pharmacists and other members of the interdisciplinary care team in nutrition management. The increased focus on assessing medications, diseases, and other issues pertinent to nutrition brings both increased opportunity and responsibility for consultant pharmacists serving long-term care facilities. PMID- 19555153 TI - Challenges in caring for aging inmates. AB - Today's prisons are vastly overcrowded and ill prepared for the fastest growing segment of the inmate population: those 50 years of age and older. Collectively, such inmates present with age-related illnesses earlier, and often their conditions are more complex than similar nonincarcerated people. Prisons focus on discipline and order; as a result, they have had to adapt to the health care needs of aging inmates, especially those with physical impairments. While some prisons have created special units for older inmates, demand for such units outpaces supply. Medication errors are common as is medication nonadherence. Continuity of care is compromised both within the system and at the point of community release. Regardless, the prison setting offers many opportunities for pharmacists to expand their practices in ways that are rewarding. PMID- 19555155 TI - Geriatric primer - common geriatric syndromes and special problems. AB - Geriatric syndromes are common problems that affect older adults. They are often thought of as causes of morbidity in one or more functional domains, but they can simultaneously be a consequence of morbidity as well. This primer will cover 12 problems commonly considered to be geriatric syndromes and highlight the potential for outcomes in one area to affect those in another. The syndromes included are: losses in activities of daily living, cognitive dysfunction, delirium versus dementia, depression, dizziness, osteoporosis, falls, sensory loss, nutrition and weight loss, pain, substance abuse, urinary incontinence, and constipation. Each syndrome is briefly discussed, followed by strategies for assessment and intervention by the pharmacist in a community setting. PMID- 19555154 TI - Racial differences in medication use among older, long-stay Veterans Affairs nursing home care unit patients. AB - OBJECTIVE: To examine racial differences in medication use by older long-stay Veterans Affairs Nursing Home Care Unit (NHCU) patients. DESIGN: Longitudinal study. SETTINGS: 133 Veterans Affairs NHCUs. PARTICIPANTS: Three thousand four hundred eighty veterans 65 years of age or older admitted between January 1, 2004, and June 30, 2005, for 90 days or more. MAIN OUTCOMES MEASURES: Prevalence of those taking nine or more medications (i.e., polypharmacy) and medications from specific therapeutic medication classes. Racial differences were determined using 0.05 level chi-squared tests. RESULTS: The sample consisted of 14.3% who were black. Blacks compared with whites (all comparisons P < 0.05 except where noted) were younger (13.6% vs. 17.4%, older than 85 years of age), had less depression (22.24 vs. 29.79%), less allergies (9.82% vs. 20.36%), and a similar rate of moderate-to severe pain (22.65% vs. 24.05; P = 0.49). The percent of polypharmacy was similar by race (blacks 74.35% vs. whites 71.18%; P = 0.62), as was the prevalence of medication class use with the exceptions that blacks were less likely than whites to take central nervous system (CNS) medications (75.75% vs. 80.14%; P = 0.02) and antihistamines (13.03% vs. 16.8%; P = 0.04). Specifically, blacks were less likely than whites to receive a selective serotonin-reuptake inhibitor (SSRI) antidepressant (20.84% vs. 27.17%; P < 0.01) or a second-generation antihistamine (3.41% vs. 6.51%; P < 0.01), but more likely than whites to receive opioids (14.63% vs. 11.27%; P = 0.03). CONCLUSION: There appears to be racial differences in the overall use of antihistamines and CNS medications and some of their subclasses. PMID- 19555156 TI - A look at Chinese pharmacies: a student's perspective. AB - As part of my fourth-year rotation, in June 2008 I had the opportunity to do my very first rotation in China and Japan as part of a foreign study program organized by Nova Southeastern University School of Pharmacy, Fort Lauderdale, Florida. Coursework would include a two-week introductory course on traditional Chinese medicine (TCM) and one week of "cultural exploration" in Japan. Fourty five students stayed at Beijing University of Chinese Medicine, International School dorms. We toured the Chinese Medicine museum and two TCM hospitals in Beijing-Pharmacology Dongfang Hospital and Xi Yuan Hospital. PMID- 19555157 TI - Reverse mentoring: an effective strategy for career growth. AB - PROBLEM: I'm beginning to feel technologically illiterate. When I see younger staff using wireless devices to access databases, update information, schedule appointments, and send messages to colleagues, I have no idea how they do so much electronically. Any suggestions? SOLUTION: Has the information age left you behind? If you were educated before 1990 you've been challenged-and probably frustrated-by rapidly evolving technology. Few older pharmacists have had formal training in using new technology, and many have taught themselves by trial and error. This is a problem in all fields. One small study found that among senior clinicians using computers, only 36% received formal course training.1 Recognizing that their senior staff risked technological illiteracy, corporate entities have addressed this problem with "reverse mentoring" programs-a rare approach in health care-but one that can propel career development. PMID- 19555158 TI - Effect of topically applied sphingomyelin-based liposomes on the ceramide level in a three-dimensional cultured human skin model. AB - Sphingomyelin-based liposomes were prepared and applied to the stratum corneum side or basal layer side of a three-dimensional (3D) cultured human skin model, and the increase in the type II ceramide (ceramide II) content of the cultured skin model was evaluated. The sphingomyelin-based liposomes were prepared by a high-pressure emulsification method, and the obtained liposomes were characterized; the particle diameter and zeta potential of the liposomes were 155.3 nm and -11.4 mV, respectively. Their spherical shape and lamella structure were observed by transmission electron microscopy. The sphingomyelin-based liposomes or saline were applied to the cultured skin model, and ceramide II was extracted from the skin model. The extracted ceramide II was separated by high performance thin-layer chromatography and quantified by a densitometer. The amount of ceramide II in the cultured skin model was significantly increased by the application of the sphingomyelin-based liposomes, compared with the nonapplication group. Thus, sphingomyelin-based liposomes are useful for enriching the ceramide level in 3D cultured skin models. PMID- 19555160 TI - Transport molecules using reverse sequence HIV-Tat polypeptides: not just any old Tat? (WO200808225). AB - BACKGROUND: Many polycationic cell penetrating peptides (CPPs), alternatively named protein transduction domains, have been used for the efficient intracellular delivery of biologically active agents. Patent WO2008/082885 relates to the properties and proposed biomedical applications of a CPP and putative-related sequences that represent the reverse sequence of a nonapeptide basic domain of the HIV-transactivator protein (Tat) (RRRQRRKKR). OBJECTIVE: This evaluation critically assesses the reported utility of such transport molecules and the numerous potential embodiments of the invention, in comparison with other recent developments in the field. We also review recent biomedical applications of Tat-derived peptide transporters. METHODS: The scope of this review includes both Tat-derived peptide transporters and other sequence-related CPPs that are polycationic in nature. RESULTS/CONCLUSION: The patent application indicates that reverse sequence HIV-Tat polypeptides can increase the transdermal delivery of an iodinated mixture of botulinum toxin, albumin and accessory proteins (Neuronox), Medy-Tox, Inc., Seoul, South Korea) as a non-covalent complex. Moreover, the invention also contemplates all variants of the reverse-sequence polypeptide and claims a variety of potential biomedical applications using the reverse sequence peptide as a delivery vector. Unfortunately, in the absence of both rigorous comparative data and toxicological analyses, it is uncertain if these transport molecules offer any advantages compared with many existing and rigorously characterised CPP vector systems. PMID- 19555159 TI - Novel therapeutics based on tau/microtubule dynamics: WO2008084483. AB - BACKGROUND: The present patent deals with the generation of peptides derived from the activity-dependent peptide and tau mimetic to study its effect on microtubule stability, its ability to bind to tubulin and MAPs, as well as promoting cell survival. OBJECTIVE: To analyze these peptides and their effects as potential therapeutic elements for neurodegenerative diseases. METHODS: We review the action of the peptides described by Gozes and collaborators and compare the effectiveness with those already reported in the literature for Alzheimer's disease. CONCLUSION: The research of Dr. Gozes and collaborators has shown that the addition of picomolar concentration of the peptides promotes cell survival, by interacting with tubulin and stabilizing the microtubules. Based on the results, these peptides seem to be very attractive candidates for therapeutical intervention in neurodegenerative diseases. PMID- 19555163 TI - Design, synthesis, and biological evaluation of 3,4,5-trimethoxyphenyl acrylamides as antinarcotic agents. AB - The synthesis and biological evaluation of 3,4,5-trimethoxyphenyl acrylamides 1a f as novel antinarcotic agents are described. The molecules were prepared by the Wittig reaction, followed by a coupling reaction between 3,4,5-trimethoxycinnamic acid (9) and aliphatic amines, which resulted in good yields. When tested for biological activity, compounds 1d-f exhibited strong inhibitory effects on the morphine withdrawal syndrome in mice due to their high binding affinities with serotonergic 5-HT1A receptors. PMID- 19555164 TI - Alpha glucosidase inhibition by stem extract of Tinospora cordifolia. AB - Inhibitors of alpha glucosidase have potential use in the treatment of diabetes mellitus. The stem extract of Tinospora cordifolia was evaluated for inhibition of the enzyme. The extract was also found to inhibit the salivary and pancreatic amylase and therefore can effectively reduce an increase in postprandial glucose level. The crude ethyl acetate, dichloromethane (DCM), chloroform and hexane extracts of Tinospora cordifolia were studied. 15 mg of the DCM extract was most effective in that showed 100 % inhibition of the alpha glucosidase whereas salivary amylase was inhibited to the extent of 75 % and pancreatic amylase to 83 %. On giving a maltose load of 2mg / g along with 0.3 mg / g body weight of the DCM Tinospora stem extract a decrease was revealed in the hyperglycemic shoot up in normal and diabetic animals by 50 and 58 % respectively as compared to the controls. The extract was found to inhibit alpha glucosidase in a non-competitive manner. PMID- 19555167 TI - Anticancer and immunomodulatory activities of novel 1,8-naphthyridine derivatives. AB - A number of 1,8-naphthyridine derivatives (22-62) have been synthesized and screened for their in vitro cytotoxicity against eight tumors and two non-tumor cell lines. Halogen substituted 1,8-naphthyridine-3-caboxamide derivatives showed potent activity with compound 47 having IC(50) of 0.41 and 0.77 microM on MIAPaCa and K-562 cancer cell lines, respectively while, compound 36 had IC(50) of 1.19 microM on PA-1 cancer cell line. However, one of the unsubstituted 1,8 naphthyridine-C-3'-heteroaryl derivative 29 showed potent cytotoxicity with IC(50) of 0.41 and 1.4 microM on PA-1 and SW620 cancer cell lines, respectively. These compounds were also evaluated for anti-inflammatory activity as suggested by downregulation of proinflammaotory cytokines. PMID- 19555169 TI - Synthesis and evaluation of in vitro antiviral activity of 2-[3-(substituted phenyl)-4,5-dihydro-1H-5-pyrazolyl]benzofuran-3-yl chloride derivatives. AB - 3-Chlorobenzofuran-2-carbaldehyde was condensed with substituted acetophenone by using the Claisen-Schmidt condensation to obtain 3-(3-chlorobenzofuran-2-yl)-1 (substituted phenyl)-2-propen-1-one (2a-m) which upon further treatment with hydrazine hydrate gave 2-[3-(substituted phenyl)-4,5-dihydro-1H-5- pyrazolyl)benzofuran-3-yl chloride derivatives (3a-m). All the newly synthesized derivatives were evaluated in vitro for cytotoxicity and antiviral activity in Crandell-Rees Feline Kidney cell, human embryonic lung (HEL) cell, HeLa cell and Vero cell cultures against different viruses. Several compounds, i.e. 2f, 2g, 2i, 2m, 3b, 3d, 3g, 3h and 3m proved quite cytotoxic to the host cells (minimum cytotoxic concentration: 1-10 microg/mL). No specific antiviral activity [50% antivirally effective concentration (EC(50)) >or= 5-fold lower than the minimum cytototoxic concentration] was observed for any of the compounds. PMID- 19555171 TI - Synthesis of imidazole-containing analogues of farnesyl pyrophosphate and evaluation of their biological activity on protein farnesyltransferase. AB - With the aim of creating new bisubstrate inhibitors of protein farnesyltransferase (FTase), new carboxylic farnesyl pyrophosphate analogues have been designed and synthesized. The original structures are built around three elements: a prenyl moiety, a 1,4-diacid motif and an imidazole ring. All the compounds were evaluated for their ability to inhibit FTase and compared with the corresponding derivatives lacking the imidazole ring, synthesized for that purpose. These new compounds are not bisubstrate inhibitors probably because the imidazole ring is not in the right position to interact with the zinc atom. However these derivatives display FPP competitive inhibition with a good activity in the carboxylic farnesyl pyrophosphate analogues series. PMID- 19555170 TI - Design, synthesis and pharmacological evaluation of 6-hydroxy-4-methylquinolin 2(1H)-one derivatives as inotropic agents. AB - Selective PDE3 inhibitors improve cardiac contractility and may be used in congestive heart failure. However, their proarrhythmic potential is the most important side effect. In this research we designed, synthesized and evaluated the potential cardiotonic activity of thirteen PDE3 inhibitors (4-[(4-methyl-2 oxo-1,2-dihydro-6-quinolinyl)oxy]butanamide analogs) using the spontaneously beating atria model. The design strategy was based on the structure of cilostamide, a selective PDE3 inhibitor. In each experiment, atrium of reserpine treated rat was isolated and the contractile and chronotropic effects of a synthetic compounds were assessed. All experiments were carried out in comparison with IBMX, amrinone and cilostamide as standard compounds. The results showed that, among the new compounds, the best pharmacological profile was obtained with the compound 6-[4-(4-methylpiperazine-1-yl)-4-oxobutoxy]-4-methylquinolin-2(1H) one, 4j, which displayed selectivity for increasing the force of contraction (165 +/- 4% change over the control) rather than the frequency rate (115 +/- 7% change over the control) at 100 microM and potent inhibitory activity of PDE3 with IC(50) = 0.20 microM. PMID- 19555172 TI - Design, synthesis and evaluation of some 1,3,5-triazinyl urea and thiourea derivatives as antimicrobial agents. AB - In an effort to establish new candidates with improved antimicrobial activities we report here the synthesis and in vitro biological evaluation of various series of compounds (5a-j) and (7a-j) which were evaluated against two Gram positive (S. aureus, B. subtilis), two Gram negative (S. typhosa, E. coli) strains and a yeast like fungi (C. albicans) using the micro-dilution procedure. Among the synthesized compounds 2-(cyclohexyl amino)-4-(3,4-dimethoxy phenyl ethyl thioureido)-6-(2-chloro phenyl ureido) s-triazine (7e) and 2-(cyclohexyl amino)-4 (3,4-dimethoxy phenyl ethyl thioureido)-6-(4-chloro phenyl ureido) s-triazine (7g) proved to be effective with MIC (0.019 mg ML(-1)) against S. typhosa & E. coli respectively. PMID- 19555173 TI - Comparative effect of N-substituted dehydroamino acids and alpha-tocopherol on rat liver lipid peroxidation activities. AB - Free radical damage has been associated with a growing number of diseases and conditions, such as autoimmune diseases, neurodegenerative disorders and multiple types of cancer. Some dehydroamino acids and corresponding peptides can function as radical scavengers. In this study the in vitro effects on rat liver lipid peroxidation levels of fourteen N-substituted dehydroamino acid derivatives and alpha-tocopherol were investigated. alpha-Tocopherol is a powerful antioxidant that is beneficial in the treatment of many free radical related diseases. The results indicated that all the compounds showed very good inhibitory effect on the lipid peroxidation compound with alpha-tocopherol at 1 mM concentrations and the inhibition rate was in the range of 70-79 % with the exception of compound 5. At 0.1 mM concentrations compounds 1, 2 and 9 were found more active than alpha tocopherol. The results confirmed that molecules such as dehydroamino acids which have reactive double bonds can act as a guard in vitro against oxidants. PMID- 19555174 TI - QSAR studies on 4-anilino-3-quinolinecarbonitriles as Src kinase inhibitors using robust PCA and both linear and nonlinear models. AB - Quantitative structure-activity relationship (QSAR) studies have been carried out on 4-anilino-3-quinolinecarbonitriles, a set of novel Src kinase inhibitors, with the aim of dissecting the structural requirements for Src inhibitory activities. After outlier identification using robust principal component analysis (robust PCA), linear models based on forward selection combined with multiple linear regression, (FS-MLR), enhanced replacement method followed by multiple linear regression (ERM) and a nonlinear model using support vector regression (SVR) were constructed and compared. All models were rigorously validated using leave-one out cross-validation (LOOCV), 5-fold cross-validations (5-CV) and shuffling external validation (SEVs). ERM seems to outperform both FS-MLR and SVR evidenced by better prediction performance (n = 35, R(2)(training) = 0.918, R(2)(pred) = 0.928). Robustness and predictive ability of ERM model were also evaluated. The generated QASR model revealed that the Src inhibitory activity of 4-anilino-3 quinolinecarbonitriles could be associated with the size of substituents in the C7 position and the steric hindrance effect. The results of the present study may be of great help in designing novel 4-anilino-3-quinolinecarbonitriles with more potent Src kinase inhibitory activity. PMID- 19555175 TI - Molecular dynamics simulation of Axillaridine-A: a potent natural cholinesterase inhibitor. AB - Molecular Dynamics (MD) simulations were carried out for human acetylcholinesterase (hAChE) and its complex with Axillaridine-A, in order to dynamically explore the active site of the protein and the behaviour of the ligand at the peripheral binding site. Simulation of the enzyme alone showed that the active site of AChE is located at the bottom of a deep and narrow cavity whose surface is lined with rings of aromatic residues while Tyr72 is almost perpendicular to the Trp286, which is responsible for stable pi -pi interactions. The complexation of AChE with Axillaridine-A, results in the reduction of gorge size due to interaction between the ligand and the active site residues. The gorge size was determined by the distance between the center of mass of Glu81 and Trp286. As far as the geometry of the active site is concerned, the presence of ligand in the active site alters its specific conformation, as revealed by stable hydrogen bondings established between amino acids. With the increasing interaction between ligand and the active amino acids, size of the active site of the complex decreases with respect to time. Axillaridine-A, forms stable pi -pi interactions with the aromatic ring of Tyr124 that results in inhibition of catalytic activity of the enzyme. This pi -pi interaction keeps the substrate stable at the edge of the catalytic gorge by inhibiting its catalytic activity. The MD results clearly provide an explanation for the binding pattern of bulky steroidal alkaloids at the active site of AChE. PMID- 19555177 TI - Aldose reductase inhibitors from Litchi chinensis Sonn. AB - Diabetes is one of the major risk factors for cataract. Aldose reductase has been reported to play an important role in sugar-induced cataract. In this study, we conducted pharmacological investigations upon experimental rat lenses using extracts of the fruits of Litchi chinensis (Sapindaceae). Of the extracts and organic fractions of L. chinensis tested, a MeOH extract and an EtOAc fraction were found to be potent inhibitors of rat lens aldose reductase (RLAR) in vitro- their IC(50) values being 3.6 and 0.3 microg/mL, respectively. From the active EtOAc fraction, four minor compounds with diverse structural moieties were isolated and identified as D-mannitol (1), 2,5-dihydroxybenzoic acid (2), delphinidin 3-O-beta-galactopyranoside-39,59-di-O-beta-glucopyranoside (3), and delphinidin 3-O-beta- galactopyranoside-39-O-beta-glucopyranoside (4). Among these, 4 was found to be the most potent RLAR inhibitor (IC(50) = 0.23 microg/mL), and may be useful in the prevention and/or treatment of diabetic complications. PMID- 19555178 TI - QSAR of aminopyrido[2,3-d]pyrimidin-7-yl derivatives: anticancer drug design by computed descriptors. AB - A series of aminopyrido[2,3-d]pyrimidin-7-yl derivatives acting as potential tyrosine kinase inhibitors having anticancer activities have been considered in the present investigation for the quantitative structure-activity relationship studies based on 2D and 3D QSAR approaches. For this purpose, various theoretical molecular descriptors were computed solely from the structures of these compounds. As the number of molecular descriptors greatly exceeds the number of observations, conventional regression does not produce reliable models and therefore, ridge regression methodology was used to solve this problem. The influence of different classes of molecular descriptors on the activity has been predicted and the most significant descriptors were obtained using the ridge regression models. Partial least squares (PLS) models were developed based on the training set for the 3D QSAR models of the above compounds. The influences of steric and electrostatic field effects generated by the contribution plots are discussed. PMID- 19555176 TI - Effect of ammonium, sodium, and potassium ions on rabbit muscle phosphofructokinase-1 and adenylate kinase activities. AB - This report shows that 30 nM PFK-1 and 30 nM AK were both affected by the presence of NH(4)(+), Na(+), and K(+) salts but with opposite consequences. Low concentrations of PFK-1 lose about half of its activity as a result of dilution and become susceptible to further activity losses owing to the presence of monovalent salts. On the other hand low concentrations of AK lose about 75 percent of its activity but regains activity losses owing to the presence of monovalent salts. It was determined that regain of AK activity did not appear to be a reflection of a major effect on the K(m) value of either AMP or ATP. Dilution to 30 nM AK resulted in no increase K(m) values compared to K(m) values at 140 nM AK. Dilution caused major decreases in the maximum velocities, V(max), when ATP or fructose 6-phosphate was the variable substrate. It was shown in earlier reports that these same low concentrations of PFK-1 and AK were susceptible inhibitions by ascorbate. These attributes are discussed as they may relate to the role of ascorbate facilitation glycogen synthesis in resting muscle and the role that the cytoskeleton infrastructure scaffold may play is also discussed. PMID- 19555181 TI - Design, synthesis and evaluation of 3-(imidazol- 1-ylmethyl)indoles as antileishmanial agents. Part II. AB - A new series of 1-benzyl-3-(imidazol-1-ylmethyl)indoles were synthesized according to a previous 3D-QSAR predictive model and assayed for their antiparasitic activity upon Leishmania mexicana promastigotes. The biological results obtained for these twelve molecules showed an IC(50) ranging from 2.3 to 32 microM and mainly illustrated the importance of the hydrophobic parameter the para-position of the benzyl group. In order to improve the activities of these compounds and to check the potential influence of the electronic parameter on this particular position, a Craig diagram was used to select original electro donating and lipophilic substituents. Synthesis and biological evaluation of ten new compounds (IC(50) between 2.5 and 5.4 microM) confirmed that only the hydrophobic field is essential for a high level of activity. PMID- 19555182 TI - Design, synthesis, spectral analysis and in vitro microbiological evaluation of 2 phenyl-3-(4,6-diarylpyrimidin-2-yl)thiazolidin-4-ones. AB - A series of novel 2-phenyl-3-(4,6-diarylpyrimidin-2-yl)thiazolidin-4-ones 23-33 were synthesized, and studied for their in vitro antibacterial and antifungal activities against clinically isolated strains. Generally compounds possessing electron donating groups showed good antibacterial activity. Compound 31, which contain both electron withdrawing chloro and electron donating methyl groups showed potent activity against all the tested Gram positive and Gram negative bacterial strains whereas compounds 32 and 33 which contain electron donating methoxy functional group at the para position of the phenyl ring attached to pyrimidine ring showed promising activity against S.aureus, S.typhii and E.coli. Compounds 32 and 33, both containing electron withdrawing groups (-Cl, -F) showed excellent activities against all the tested A. flavus, Mucor, Rhizopus and M.gypsuem fungal strains. while against Mucor, compound 27 which contains an electron donating methyl group at the para position of the phenyl ring attached to pyrimidine ring showed promising activity. Also compound 31, which contains both electron withdrawing chloro and electron donating methyl groups showed potent activity against A. flavus and Rhizopus. PMID- 19555184 TI - Synthesis, inhibition of NO production and antiproliferative activities of some indole derivatives. AB - The synthesis and the biological evaluation of pyrano[3,2-e]indoles and their reaction intermediates are described. The compounds prepared were evaluated for their inhibition of NO production, antioxidant activity and also for their ability to inhibit in vitro the growth of four human tumor cell lines: large lung carcinoma (COR-L23), alveolar basal epithelial carcinoma (A549), amelanotic melanoma (C32) and melanoma (A375). The two reaction intermediates, 5a and 5b, showed the highest inhibition of NO production in murine monocytic macrophage (IC(50) = 1.1 microM and IC(50) = 2.3 microM respectively). Compound 5a was the most active against melanotic melanoma (IC(50) = 11.8 microM) while the other compounds exhibited weak cytotoxicity with IC(50) values >50 microM on all cell lines. PMID- 19555186 TI - Compounds with neuroprotective activity from the medicinal plant Machilus thunbergii. AB - The dichloromethane fraction of the bark of Machilus thunbergii Sieb. et Zucc. (Lauraceae) significantly protected primary cultures of rat cortical cells exposed to the excitotoxic amino acid, L-glutamate. Through the activity-guided isolation from the CH(2)Cl(2) fraction, (+)-9'-hydroxygalbelgin (1), isogalcatin B (2), (7S,8S,8'R)-3',4'-dimethoxy-3,4,-methylenedioxylignan-7-ol (3), 1-hydroxy 7-hydroxymethyl-6-methoxyxanthone (4), 5,7-dimethoxy-3',4'-methylenedioxyflavan-3 ol (5), (+)-(3S,4S,6R)-3,6-dihydroxypiperitone (6), protocatechuic acid methyl ester (7) and tyrosol (8) were obtained. All of them had significant neuroprotective activities against glutamate-induced neurotoxicity in primary cultures of rat cortical cells at concentrations ranging from 0.1 microM to 10.0 microM and were comparable to MK-801, a well-known inhibitor of glutamate receptor. PMID- 19555185 TI - Dual effects of aliphatic carboxylic acids on cresolase and catecholase reactions of mushroom tyrosinase. AB - Catecholase and cresolase activities of mushroom tyrosinase (MT) were studied in presence of some n-alkyl carboxylic acid derivatives. Catecholase activity of MT achieved its optimal activity in presence of 1.0, 1.25, 2.0, 2.2 and 3.2 mM of pyruvic acid, acrylic acid, propanoic acid, 2-oxo-butanoic acid, and 2-oxo octanoic acid, respectively. Contrarily, the cresolase activity of MT was inhibited by all type of the above acids. Propanoic acid caused an uncompetitive mode of inhibition (K(i)=0.14 mM), however, the pyruvic, acrylic, 2-oxo-butanoic and 2-oxo-octanoic acids showed a competitive manner of inhibition with the inhibition constants (K(i)) of 0.36, 0.6, 3.6 and 4.5 mM, respectively. So, it seems that, there is a physical difference in the docking of mono- and o diphenols to the tyrosinase active site. This difference could be an essential determinant for the course of the catalytic cycle. Monophenols are proposed to bind only the oxyform of the tyrosinase. It is likely that the binding of acids occurs through their carboxylate group with one copper ion of the binuclear site. Thus, they could completely block the cresolase reaction, by preventing monophenol binding to the enzyme. From an allosteric point of view, n-alkyl acids may be involved in activation of MT catecholase reactions. PMID- 19555187 TI - The pyrostatins A and B do not inhibit N-acetyl-beta-D-glucosaminidase. AB - The compounds pyrostatin A and B, isolated from Streptomyces sp. SA-3501 have been reported as N-acetyl-beta-D-glucosaminidase inhibitors with inhibition constants in the micromolar range. Recently, a comparison of NMR spectral data of the pyrostatins has led to a structural revision of the pyrostatins. It was shown that the pyrostatins A and B are identical to the ectoines 5-hydroxectoine and ectoine, respectively. Ectoines are known as compatible osmolytes in many halophilic and stress-tolerant bacteria. We have performed enzymatic experiments demonstrating that neither ectoine nor 5-hydroxyectoine exhibit an inhibitory effect on N-acetyl-beta-D-glucosaminidase. The previously reported inhibition of N-acetyl-beta-D-glucosaminidase by pyrostatins A and B may thus be due to the contamination of the compound preparations with a strong N-acetyl-beta-D glucosaminidase inhibitor with an inhibition constant (Ki) in the nanomolar range, as has been reported in other Streptomyces species. PMID- 19555191 TI - Potentiation of murine innate immunity by alpha-galacturonosyl-type glycosphingolipids isolated from Sphingomonas yanoikuyae and S. terrae. AB - Glycosphingolipids (GSLs) are components of the outer membrane of Sphingomonas species, commonly classified into two types, alpha-glucuronosyl ceramide (alpha GlcACer) and alpha-galacturonosyl ceramide (alpha-GalACer), respectively. GSL-7 from S. yanoikuyae and GSL-13 from S. terrae, with alpha-GalACer-type structure, possess dihydrosphingosine but with a different ratio of C21cyclopropane to C20:1, while other parts remain similar. We therefore examined if this difference in the ratio of C21cyclopropane to C20:1 in the two ceramides may influence activation of, not only invariant natural killer T (iNKT) cells, but also other cells involved in innate immunity. GSL-7 with a large proportion of C21cyclopropane induced stronger activation of iNKT cells, natural killer cells, dendritic cells, and macrophages than GSL-13 with a large proportion of C20:1. The results show that a higher ratio of C21cyclopropane to C20:1 in the dihydrosphingosine molecule allows a more optimal activation of iNKT cells and other cell types. PMID- 19555194 TI - Aluminum, calcium ion and radiofrequency synergism in acceleration of lymphomagenesis. AB - This study that was done on lymphomagene-bearing mice indicates a synergism aluminum-radiofrequency which induces an early increase in mortality that is in concomitance with lymphoid elements proliferation and infiltration of spleen and liver. These two last phenomena were assessed by determination of the hypertrophic index (Growth Index) which is the organ weight to to the body weight ratio, as well as by the histopathological examination of the organ tissue. The importance of this synergism appears to be determined by the ionization at the physiological pH of the used aluminum complexes: much higher with lactate complex than with the citrate one. On the other hand, this dissociation appears to induce a remarkable acceleration of the mortality and the lymphoid elements-related hypertrophy of the spleen and liver at early age. Aluminum complexes are known as modifiers of the intracellular calcium homeostasis, and to verify if such process could be implicated in this synergism, the effects of calcium chloride were assayed, in this case the calcium-overload had no effects in the presence of a workable cellular control of intracellular calcium homeostasis. This finding support the hypothesis that ionized aluminum provided by lactate may be implicated in the inhibition of the buffering and extruding extracellular calcium system. PMID- 19555197 TI - Effect of HMG-CoA reductase inhibitors on activation of human gammadeltaT cells induced by Mycobacterium tuberculosis antigens. AB - Lipid rafts are cholesterol-enriched microdomains which act as a platform for the initiation of T-cell activation. To investigate effect of endogenous cholesterol on lipid rafts formation and activation of gammadeltaT cells, human peripheral blood mononuclear cells were stimulated in vitro with Mycobacterium tuberculosis antigens (Mtb-Ag). Lovastatin and fluvastatin, two 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (HMGCR) inhibitors, were used to block endogenous cholesterol biosynthesis. The expression of ganglioside GM1 (GM1), a lipid rafts marker, and CD69, an activation marker, and the level of tyrosine phosphorylation in gammadeltaT cells were measured by flow cytometry. The expression and aggregation of GM1 were also detected with laser confocal microscopy. We found that lovastatin and fluvastatin could obviously inhibit tyrosine phosphorylation and expression of GM1 and CD69 in gammadeltaT cells induced by Mtb-Ag. These results collectively indicated that HMGCR inhibitors might interfere with the formation of lipid rafts and inhibit the activation of gammadeltaT cells induced by Mtb-Ag. PMID- 19555200 TI - Immunomodulatory activity of xanthohumol: inhibition of T cell proliferation, cell-mediated cytotoxicity and Th1 cytokine production through suppression of NF kappaB. AB - Xanthohumol (XN), a prenylated chalcone present in hops (Humulus lupus L.) and beer, exhibits anti-inflammatory, antioxidant and antiproliferative activity, but has not been studied for effects on T cell-mediated immune responses. Here we demonstrate that XN has profound immunosuppressive effects on T cell proliferation, development of IL-2 activated killer (LAK) cells, cytotoxic T lymphocytes (CTLs), and production of Th1 cytokines (IL-2, IFN-gamma and TNF alpha). The suppression of these cell-mediated immune responses by XN was at, least in part, due to the inhibition of nuclear factor kappa B (NF-kappaB) transcription factor through suppression of phosphorylation of IkappaBalpha, an inhibitor of NF-kappaB. PMID- 19555203 TI - Immunomodulatory activities of Punarnavine, an alkaloid from Boerhaavia diffusa. AB - The effect of Punarnavine on the immune system was studied using Balb/c mice. Intraperitoneal administration of Punarnavine (40 mg/kg body weight) was found to enhance the total WBC count on 6(th) day. Bone marrow cellularity and number of alpha-esterase positive cells were also increased by the administration of Punarnavine. Treatment of Punarnavine along with the antigen, sheep red blood cells (SRBC), produced an enhancement in the circulating antibody titer and the number of plaque forming cells (PFC) in the spleen. Maximum number of PFC was obtained on the 6(th) day. Punarnavine also showed enhanced proliferation of splenocytes, thymocytes and bone marrow cells both in the presence and absence of specific mitogens in vitro and in vivo. More over administration of Punarnavine significantly reduced the LPS induced elevated levels of proinflammatory cytokines such as TNF-alpha, IL-1beta, and IL-6 in mice. These results indicate the immunomodulatory activity of Punarnavine. PMID- 19555204 TI - Biochemical and cellular toxicology of peroxynitrite: implications in cell death and autoimmune phenomenon. AB - Reactive nitrogen species include nitric oxide (.NO), peroxynitrite (ONOO(-)) and nitrogen dioxide radical (NO2*). Peroxynitrite is a reactive oxidant, produced from nitric oxide (*NO) and superoxide anion (O(2*-), that reacts with a variety of biological macromolecules. It is produced in the body in response to physiological stress and environmental toxins. It is a potent trigger of oxidative protein and DNA damage-including DNA strand breakage and base modification. It activates the nuclear enzyme poly-ADP ribose polymerase (PARP) resulting in energy depletion and apoptosis/necrosis of cells. Peroxynitrite generation is a crucial pathological mechanism in stroke, diabetes, inflammation, neurodegeneration, cancer, etc. Peroxynitrite modified DNA may also lead to the generation of autoantibodies in various autoimmune disorders such as systemic lupus erythematosus (SLE). In chronic inflammatory diseases, peroxynitrite formed by phagocytic cells may cause damage to DNA, generating neoepitopes leading to the production of autoantibodies. Hence, understanding the pathophysiology of peroxynitrite could lead to important therapeutic interventions. PMID- 19555205 TI - Improved immunogenicity and protective efficacy of a replicon DNA vaccine encoding the Hc domain of botulinum neurotoxin serotype A by electric pulses and protein boosting. AB - Vaccination by intramuscular injection of naked DNA is very efficient in mouse model, but immunogenicity of DNA vaccines needs to be improved in human use. Thus, we wanted to determine whether suitable electric pulses-mediated DNA delivery technology and DNA prime-protein boost regimen could improve the immunogenicity and protective efficacy of the replicon DNA vaccine pSCARSHc in mouse model. In this study, the immunogenicity and protective efficacy of the replicon DNA vaccine pSCARSHc following electric pulses were dramatically improved in Balb/c mice. Also, priming of the immune response by DNA vaccination followed by a single booster with AHc protein immunogen resulted in very high levels of ELISA and neutralization antibodies and afforded more efficient protection against botulinum neurotoxin serotype A. Therefore, these methods described here potentially provide suitable strategies in developing an efficacious vaccine against Clostridium botulinum neurotoxin serotype A. PMID- 19555207 TI - Simultaneous detection of five biothreat agents in powder samples by a multiplexed suspension array. AB - A suspension array-based multiplexed immunoassay was developed for rapid, sensitive, specific, and simultaneous detection of multiple biothreat-associated agents in powder samples. The 5-plexed immunoassays using sets of 9-plexed coupled fluorescent beads were employed to simultaneously detect five representative biothreat agents, including B. anthracis spore, Y. pestis, SARS CoV, staphylococcal enterotoxin B (SEB) and ricin from a single powder sample and the feasibility for field samples was demonstrated by both blinded and standard laboratory trials. The detection sensitivity and dynamic range for the five biothreat agents from different powders might be varied depending on the nature of the powder and the feature of the contaminating agent. The limit of detection for Y. pestis, B. anthracis spores, SEB, ricin, SARS-CoV N protein in milk powder was 20 cfu, 111 cfu, 110pg, 5.4 ng and 2 ng per test respectively. Compared to conventional ELISA method, the suspension array has a higher sensitive ability, and can detect five biothreat agents simultaneously with high reproducibility. PMID- 19555208 TI - Zoledronic acid, an aminobisphosphonate, modulates differentiation and maturation of human dendritic cells. AB - Zoledronic acid (ZOL), an effective nitrogen-containing bisphosphonate against excessive bone loss, has been shown affecting the function of cells of both innate and acquired immunity. In this study, we tested the effect of ZOL on differentiation and maturation of human myeloid dendritic cells (DC). When ZOL (1.1 to 10 microM) was added to the culture of starting monocytes, but not to immature DC, the recovery rate of DC was markedly reduced in a concentration dependent manner. The mature DC differentiated in the presence of ZOL had fewer and shorter cell projections. ZOL treatment affected DC differentiation and maturation in terms of lower expression of CD1a, CD11c, CD83, CD86, DC-SIGN, HLA DR, and, in contrast, higher expression of CD80. IL-10 production by DC was inhibited by ZOL treatment whereas IL-12p70 secretion remained unchanged. Interestingly, ZOL augmented the allostimulatory activity of DC on naive CD4(++)CD45(+)RA(++) T cells in terms of their proliferation and interferon-gamma production. Addition of geranylgeraniol abrogated the effect of ZOL on DC differentiation and prenylation of Rap1A. It suggests that ZOL redirects DC differentiation toward a state of atypical maturation with allostimulatory function and this effect may go through prevention of Rap1A prenylation. PMID- 19555209 TI - The potential use of Toll-like receptor (TLR) agonists and antagonists as prophylactic and/or therapeutic agents. AB - Toll-like receptors (TLR) and their ligands are one of the main players in the initiation of innate immunity which precedes, and is required, for the establishment of adaptive immunity. Manipulating the immune response by using TLR agonists or antagonists might be of therapeutic and/or prophylactic value. This review covers; 1-TLR. their natural ligands and ligand - TLR signaling events, 2 TLR against and their use in clinical trials as vaccine adjuvants, and to treat allergy, cancer and infectious diseases, 3-TLR antagonists and their use in clinical trials to treat septic shock and autoimmune diseases. Potential drawbacks related to their potential use as prophylactic and/or therapeutic agents are discussed. PMID- 19555210 TI - Growth-inhibitory effects of a Bulnesia sarmienti aqueous extract on A549 cells in vitro and S180 cells in vivo. AB - The effects of Bulnesia sarmienti (BS) aqueous extract on the cell growth of A549 cell lines were investigated. BS has strong cytotoxic activity on the A549 cell lines (IC(50); less than 100 microg/mL) in MTT assay. HPLC confirmed that BS contains catechins as major compound. Cell cycle analysis by flow cytometry indicated that BS arrested the cell cycle in the sub-G(1) phase. BS induced DNA fragmentation, and increased the expression of the p53 protein in immunoblot analysis. These results indicated that the anticancer effect of BS was mediated via the process of apoptosis and growth-inhibition. PMID- 19555215 TI - Intranasal immunization with Porphyromonas gingivalis and atherosclerosis. AB - Periodontal disease is a highly prevalent disorder affecting up to 90% of the global population. Recent epidemiological studies have shown that an association exists between periodontal disease and cardiovascular disease. Porphyromonas gingivalis, the causative agent of destructive chronic inflammation in the periodontium, can accelerate atheroma deposition in animal models. Emerging evidence suggests that vaccination against this pathogen's virulence factors may confer disease resistance. In this review, we focus on the role of inflammatory mechanisms in the formation and activation of atherosclerotic plaques accelerated by P. gingivalis in an apo E-deficient mouse model. Further, we examine whether a nasal vaccine-induced antigen-specific mucosal response can reduce P. gingivalis accelerated atherosclerosis. PMID- 19555216 TI - Identification of an HLA-A*0201-restrictive CTL epitope from MUC4 for applicable vaccine therapy. AB - Recent research has indicated that MUC4 plays an important role in the development of many tumors and may prove useful as a novel cancer immunotherapy target. We aimed to identify HLA-A*0201-restrictive cytotoxic T lymphocyte (CTL) epitopes of the cancer-associated antigen MUC4. The MUC4 sequence was scanned for immunogenic peptides using HLA-binding prediction software. Dendritic cells (DCs) from peripheral blood mononuclear cells (PBMCs) were induced by cytokines. Five possible CTL epitopes were selected by software analysis, synthesized, and used to pulse mature DCs. The CD8(+) T cells from PBMCs from an HLA-A*0201 healthy donor were stimulated with autologous MUC4-peptide-loaded DCs and expanded in vitro. T cell activation was assessed by ELISPOT, and cytotoxicity was determined by (51)chromium ((51)Cr)-release assays. Our results show that CTLs induced by peptide P01204 could lyse T2 cells pulsed with peptide P01204 and HCT-116 cells (MUC4(+), HLA-A2(+)). Compared with a control peptide, P01204 increased the number of IFN-gamma producing T cells. Overall, these results suggest that P01204 is a novel HLA-A*0201-restrictive CTL epitope of the cancer-associated antigen MUC4. This will provide a foundation for the development of tumor-specific peptide vaccines. PMID- 19555219 TI - Upper airways sensory irritation responses of mice exposed to mainstream smoke from four cigarette types. AB - Relative sensory irritation responses for Swiss-Webster mice exposed nose-only to mainstream tobacco smoke were evaluated for several cigarette types using a smoking regimen consisting of a 35-ml puff, 2 s in duration, taken once per minute. The degree of sensory irritation for each cigarette type was evaluated as the smoke concentration inducing a 50% reduction in breathing frequency. The smoke concentration inducing 50% respiratory depression is called the RD(50) value. Study findings suggest that mainstream tobacco smoke from the Eclipse cigarette, which primarily heats rather than burns tobacco, yielded an RD(50) that was significantly higher (approximately twofold) than a tobacco-burning leading ultralight or the 2R4F or 1R5F reference cigarettes. This is indicative of reduced upper airways irritation by Eclipse that may be due to its distinct design. Study findings suggest that the irritating nature of mainstream tobacco smoke from different cigarette types can be evaluated effectively in terms of smoke concentration using the relative sensory irritation assessment. These findings constitute the first report about use of the RD(50) sensory irritation response during comparative evaluations of mainstream tobacco smoke. PMID- 19555217 TI - Immune stimulatory effects of Loranthi ramulus on macrophages through the increase of NO and TNF-alpha. AB - The activation of macrophages by microorganisms plays an important role in host defense and immunopathology. Loranthi ramulus (LR) is commonly used as a traditional drug and health food in Korea. Here, we investigated the regulatory effects of LR on macrophage-mediated immune responses. Treatment of macrophages with LR resulted in the enhanced cell-surface expression of CD80, CD86 and major histocompatibility complex (MHC) class II, as well as the enhanced production of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha, and also iNOS and TNF alpha mRNA expression. These alterations of LR-treated cells were associated with the activation of NF-kappaB and mitogen-activated protein kinases (MAPKs). LR increased the phosphorylation of MAPKs (JNK, ERK1/2, p38 MAPK) and the activation of NF-kappaB in Raw 264.7 cells. These results suggest that LR has increased NO and TNF-alpha production through phosphorylation of all three MAPKs following IkappaBalpha degradation and NF-kappaB activation. In conclusion, our results demonstrate that LR can effectively promote the activation of macrophages, suggesting that LR may possess the potential to regulate immune responses. PMID- 19555222 TI - Effect of filtration by activated charcoal on the toxicological activity of cigarette mainstream smoke from experimental cigarettes. AB - Activated charcoal (AC) filtration reportedly decreases the yields of smoke vapor phase constituents including some identified as human carcinogens and respiratory irritants. Non-clinical studies including chemical smoke analysis, in vitro cytotoxicity and mutagenicity (bacterial and mammalian cells), and in vivo subchronic rat inhalation studies were carried out using machine smoking at ISO conditions with lit-end research cigarettes containing AC filters. The objective was to assess whether AC filter technology would alter the established toxicity profile of mainstream smoke by increasing or decreasing any known toxicological properties, or elicit new ones. The reduced yield of vapor phase irritants from AC filter cigarettes correlated with markedly decreased in vitro cytotoxicity and in vivo morphology of the nose and lower respiratory tract. Increased yields of particulate phase constituents (e.g. polycyclic aromatic hydrocarbons) in AC filtered smoke were noted in comparison to controls in some studies. The in vitro bacterial mutagenicity of AC filtered smoke particulate preparations was occasionally increased over control levels. Laryngeal epithelial thickness was increased in some rats inhaling AC filtered smoke in comparison to controls, an effect perhaps related to higher inspiratory flow. When tested under more intense Massachusetts Department of Public Health smoking conditions, AC filter associated reductions in vapor phase constituent yields were smaller than those seen with ISO conditions, but the effect on in vitro cytotoxicity remained. PMID- 19555223 TI - Evaluation of a novel inhalation exposure system to determine acute respiratory responses to tobacco and polymer pyrolysate mixtures in Swiss-Webster mice. AB - Modern cigarette production processes are highly automated and yield millions of cigarettes per day. The forming cigarette and its components contact many different materials in the production process, some of which may leave minute residues. The potential for small inclusions of non-cigarette materials such as wood, plastic, cardboard and other materials exists from the bulk handling and processing of tobacco, in spite of vigilant workers and numerous online systems designed to keep the tobacco stream clean. Currently, there are no published methods that describe an approach to evaluate the potential toxicological impact of these non-tobacco residues and inclusions on the biological activity from exposure to the complex mixture of tobacco smoke. There are, however, many methods which describe toxicological evaluation approaches for pure materials, particularly synthetic polymers. We used the Deutsche Institute fur Normung (DIN) 53-436 tube furnace and nose-only exposure chamber in combination to conduct pilot studies in Swiss-Webster mice in order to develop a standardized methodology for the evaluation of sensory irritation and other potentially useful biological endpoints for predicting any potential hazards. Sensory and/or pulmonary irritation was assessed based on respiratory function parameters using the ASTM E981-84 method described by Alarie (1966) in mice, exposed to test atmospheres of 100% tobacco pyrolysate or tobacco/polymer pyrolysate mixtures. Other biological evaluations included respiratory function parameters, clinical signs, body weights, bronchoalveolar lavage fluid analysis, carboxyhemoglobin, blood cyanide concentrations and histopathology of the respiratory tract. These pilot studies have demonstrated that such an approach can detect biological changes resulting from exposure to unique tobacco/polymer pyrolysates. Small differences were detected in the sensory irritation responses (respiratory function), activation state of pulmonary macrophages, and histopathological findings in the nose of mice exposed to 100% tobacco or tobacco/polymer pyrolysates. Analytical measurements were also performed in order to characterize the test atmospheric changes that could occur from inclusion of the polymer into the tobacco. These included DIN-generated wet total particulate matter (TPM) DIN Generated wet TPM (DWTPM), nicotine, cyanide, formaldehyde, acetaldehyde, acrolein, carbon monoxide, carbon dioxide, and nitrogen oxides. We attempted to correlate analyte differences in the test atmospheres with the resulting biological findings in the mice. The results demonstrated that this approach could detect minimal toxicological effects in mice exposed to test atmospheres of 100% tobacco or 70%/30% tobacco/polymer pyrolysates. PMID- 19555224 TI - Temporal integration in nasal lateralization of homologous propionates. AB - For nasal irritation from volatile chemicals, a version of Haber's rule (k = C(n)T) can model the trade-off between concentration (C) and duration of exposure (T) to achieve a fixed sensory impact, e.g. threshold-level irritation or a fixed suprathreshold intensity. The term k is a constant. The exponent, n, represents how well the system integrates over time. An exponent of 1 indicates complete temporal integration: an x-fold increase in stimulus duration exactly compensates for cutting the concentration 1/x. An exponent greater than 1 indicates incomplete temporal integration: more than an x-fold increase in duration is needed. In a previous study of homologous alcohols, n varied systematically with number of methylene units: integration became more complete as the length of the carbon chain increased. To explore the generality of this finding, we tested homologous esters that differ in the number of methylene units: n-ethyl propionate, n-propyl propionate, and n-butyl propionate. Nasal lateralization was used to measure irritation thresholds. Human subjects received a fixed concentration of a single compound within each experimental session. Stimulus duration was varied to find the briefest stimulus that caused lateralizable irritation. Concentration and compound varied across sessions. Consistent with results with n-alcohols, integration became more complete as the number of methylene units increased. Lipid solubility varies with chain length; hence, solubility in the nasal mucosa may play a role in the dynamics of irritation. Further, preliminary analyses suggest that, for data pooled across both chemical series, n varies systematically with molecular parameters related to solubility and diffusion. PMID- 19555226 TI - Increased nasal epithelial ciliary beat frequency associated with lifestyle tobacco smoke exposure. AB - The ciliated epithelium of the respiratory airways is one of the first vital systemic surfaces in contact with the ambient air. Ex vivo nasal epithelial ciliary beat frequency (CBF) at room temperature is on the order of 7-8 Hz but may be stimulated by irritant exposure. The upregulation of CBF in response to acute irritant exposure is generally considered to be a transient event with eventual return to baseline. However, studies of CBF dynamics in response to typical lifestyle exposures are limited. This study assessed nasal epithelial CBF among human subjects as a function of quantifiable lifestyle tobacco smoke exposure. Nasal epithelial biopsies were obtained from human subjects with well documented histories of tobacco smoke exposure. CBF was determined using a digital photometric technique and concurrent assays of nasal nitric oxide and urine cotinine and creatinine were performed. Mean CBF among active smokers and non-smokers exposed to environmental tobacco smoke (ETS) was elevated over non smokers. Although there were dramatic differences in relative levels of tobacco smoke exposure, CBF values among tobacco smoke-exposed groups were comparable. Parallel in vitro studies of cultured nasal epithelium exposed to cigarette smoke condensate further supported these observations. These studies suggest that persistent elevation in nasal epithelial CBF is an early, subtle, physiologic effect associated with lifestyle tobacco smoke exposure. The molecular mechanisms that upregulate CBF may also create a cell molecular milieu capable of provoking the eventual emergence of more overt adverse health effects and the pathogenesis of chronic airway disease. PMID- 19555227 TI - Effects of biomass combustion smoke on hematological and antioxidant profile among children (8-13 years) in India. AB - The use of solid biomass fuel in traditional stoves has been associated with respiratory symptoms of chronic airway inflammation and higher rates of respiratory infections. The mechanisms of such associations remain unclear. In this study we examine the association between exposures to indoor pollution and the hematological and antioxidant profile in children. We found increases in the respiratory symptoms "cough without cold" [odds ratio (OR) 4.27; 95% confidence interval (CI) 3.00-4.95], "cough in the morning" (OR 3.80; CI 2.40-5.15), and "wheeze" (OR 3.60; CI 2.75-5.0) in children living in homes used solid biomass for cooking after adjustment for potential confounder variables. Children who lived in the households that cook with traditional biomass fuels had low hemoglobin and red blood cell (RBC) values, but raised white blood cell (WBC), neutrophil, and eosinophil counts. Furthermore, we examined the ascorbic acid, superoxide dismutase (SOD), and glutathione/oxidized glutathione (GSH/GSSG) ratio levels in the blood of the subjects and observed low levels of ascorbic acid (AA), SOD, and GSH/GSSG ratio in children lived in biomass used households. These findings suggest that the biomass smoke has the potential to produce oxidative stress and adverse health effects in children. There is much more work needed to confirm our conclusions. Investigating the mechanisms underlying air pollution induced health problems would allow a more targeted approach to remove the most toxic components of air pollution, and could possibly provide a means to decrease individual sensitivity to air pollution. PMID- 19555228 TI - Synergistic effect of air pollution and habitual smoking on the retention of inorganic fibers identified as ferruginous bodies in autopsy cases in Mexico City. AB - In order to evaluate the synergistic effect of habitual smoking and air pollution in Mexico City on the retention of inorganic fibers, ferruginous bodies (FB) were quantified as markers of exposure to inorganic fibers in lung digests from 426 autopsy cases. FB were isolated from 426 lung digests from cases with several lung diseases. The results revealed more retention of FB in the smokers group than in non-smokers: 38 FB per gram (FB/g) versus 11.2 FB/g, respectively (p < 0.05). Male smokers living in Mexico City increased their median to 54 FB/g. This contrasts with the median of outside residents: 11.2 FB/g (p < 0.002). Housewives and manual laborers increased their medians when the smoking habit was positive: from 11 to 14 FB/g, and from 16 to 21.5 FB/g, respectively. There is an effect of tobacco smoke on the retention of more fibers identified as FB when the individuals are males and Mexico City residents. PMID- 19555225 TI - Modulation of ozone-sensitive genes in alpha-tocopherol transfer protein null mice. AB - Alpha-tocopherol transfer protein (ATTP) null mice (ATTP-/-) have a systemic alpha-tocopherol (AT) deficiency, with their lung AT levels being < 10% of those in AT-replete ATTP(+/+) mice when fed a standard rodent chow diet. ATTP(+/+) and ATTP(-/-) mice (4 wk old male mice, n = 16 per group) were fed a standard diet (35 IU AT/kg diet) for 8-12 wk, exposed 6 h/day for 3 days to either to O(3) (0.5 ppm) or filtered air, then sacrificed. No significant differences in plasma or lung AT concentrations were observed in response to this level of O(3) exposure. Lung genomic responses of the lungs to O(3) were determined using Affymetrix 430A 2.0 arrays containing over 22,600 probe sets representing 14,000 well characterized mouse genes. As compared with filtered air exposure, O(3) exposure resulted in 99 genes being differentially expressed in ATTP(-/-) mice, as compared to 52 differentially expressed genes in ATTP(+/+) mice. The data revealed an O(3)-induced upregulation of genes related to cell proliferation/DNA repair and inflammatory-immune responses in both ATTP(+/+) and ATTP(-/-) mice, with the expression of 22 genes being common to both, whereas 30 and 77 genes were unique to ATTP(+/+) and ATTP(-/-) mice, respectively. The expressions of O(3) sensitive genes-Timp1, Areg, Birc5 and Tnc-were seen to be further modulated by AT status. The present study reveals AT modulation of adaptive response of lung genome to O(3) exposure. PMID- 19555229 TI - Chemicals present in automobile traffic tunnels and the possible community health hazards: a review of the literature. AB - Dozens of volatile and semivolatile organic compounds can be detected in vehicle exhaust, along with numerous metals and oxides of sulfur, nitrogen, and carbon. While the adverse effects of these chemicals have been extensively studied surrounding open roadways, the hazards to local residents and commuters resulting from the presence of tunnel emission chemicals are less well known. Commuters and workers within tunnels are also exposed to tunnel atmospheres, and the risks have only been evaluated to a limited extent. Approximately 50 studies conducted at more than 35 different international traffic tunnels were reviewed in order to characterize the potential health impact on individuals residing near these tunnels. One objective of this article is to identify those chemicals that deserve further study in order to understand the hazards to humans who work in these tunnels, as well as the risks to those in the surrounding community. The second objective is to present the available information regarding the hazards to those living near these tunnels. The published information, for the most part, indicates that the concentration of most toxicants detected in communities exposed to tunnel emissions are below those concentrations that are generally considered to pose either a significant acute or chronic health hazard. However, there have been no comprehensive studies that have evaluated the concentration of all of the relevant toxicants on a real-time basis or using repetitive time weighted average sampling. Based on our analysis of the existing information appearing in peer-reviewed literature and government reports, additional information on the variation of concentrations of various chemicals over time near the tunnel exits would be helpful. Optimally, these would be better if evaluated in conjunction with traffic magnitude and vehicle type. It would also be useful to further characterize acute exposures to commuters or tunnel workers during times of heavy volume or slow-moving traffic due to accidents within the tunnel structure, when tunnel pollutant levels would be expected to be substantially elevated. A recent review by the Australia's National Health and Medical Research Council also discusses tunnel and air quality in detail (2008). Nearly 300 references are cited. PMID- 19555230 TI - Computer controlled multi-walled carbon nanotube inhalation exposure system. AB - Inhalation exposure systems are necessary tools for determining the dose-response relationship of inhaled toxicants under a variety of exposure conditions. The objective of this project was to develop an automated computer controlled system to expose small laboratory animals to precise concentrations of airborne multi walled carbon nanotubes (MWCNT). An aerosol generator was developed which was capable of suspending a respirable fraction of multi-walled carbon nanotubes from bulk material. The output of the generator was used to expose small laboratory animals to constant aerosol concentrations up to 12 mg/m(3). Particle distribution and morphology of the MWCNT aerosol delivered to the exposure chamber were measured and compared to samples previously taken from air inside a facility that produces MWCNT. The comparison showed the MWCNT generator was producing particles similar in size and shape to those found in a work environment. The inhalation exposure system combined air flow controllers, particle monitors, data acquisition devices, and custom software with automatic feedback control to achieve constant and repeatable exposure chamber temperature, relative humidity, pressure, aerosol concentration, and particle size distribution. The automatic control algorithm was capable of maintaining the mean aerosol concentration to within 0.1 mg/m(3) of the selected target value, and it could reach 95% of the target value in less than 10 minutes during the start-up of an inhalation exposure. One of the major advantages of this system was that once the exposure parameters were selected, a minimum amount of operator intervention was required over the exposure period. PMID- 19555232 TI - The effect of resveratrol in tracheal tissue of rats exposed to cigarette smoke. AB - OBJECTIVE: The aim of this study was to evaluate the effect of resveratrol on the tracheal tissue of rats exposed to cigarette smoke. MATERIALS AND METHODS: 40 adult Wistar albino rats were divided into four groups for an experiment of 6 weeks. Animals in group 1 were controls (n = 10). Rats in group 2 were exposed to cigarette smoke only, and rats in group 3 received daily intraperitoneal injections of resveratrol (10 mg/kg/d). Animals in group 4 were exposed to both cigarette smoke and intraperitoneal injections of resveratrol. Rats of all groups were sacrificed using cervical dislocation. The tracheas were removed and embedded in paraffin blocks. Sections of 4-5 mum thickness were prepared from the blocks. These sections were stained with hematoxylin and eosin, periodic acid Schiff, and Alcian blue and viewed with a Leica DFC 280 light microscope. RESULTS: Tracheal sections showed that, in group 2 (cigarette smoke group), there was desquamation of epithelial cells into the tracheal lumen, loss of cilia in the epithelial layer, an increase of goblet cells, activation of serous glands at the submucosa, and cell infiltration. In group 4 (cigarette smoke + resveratrol group), all these findings also existed but only a few sections were affected. It was observed that cigarette smoking caused morphological changes such as epithelial degeneration in the upper airway. These morphological changes were correlated with the amount of toxic substances in the cigarette smoke. CONCLUSION: We found that resveratrol had a preventive role in the histopathological changes caused by cigarette smoking in the rat trachea. PMID- 19555236 TI - Generic omeprazole delayed-release capsules: in vitro performance evaluations. AB - BACKGROUND: After the patent on omeprazole delayed-release capsules expired, Food and Drug Administration (FDA) approved several generic omeprazole delayed-release capsule applications. FDA has received some complaints concerning a lack of therapeutic effect of the generic omeprazole delayed-release capsules. AIM: To investigate the quality of five different marketed generic omeprazole delayed release capsules. METHOD: The dissolution characteristics of these generic omeprazole delayed-release capsules were determined according to the United States Pharmacopeia (USP). Additional dissolution studies under simulated in vivo physiological conditions were also conducted to determine whether generic omeprazole capsules would perform similarly under these conditions. RESULTS: The experimental data show that all the generic omeprazole delayed-release capsules met the USP standards. The in vitro dissolution of generic drugs is similar to that of the brand omeprazole product. CONCLUSIONS: There is no scientific evidence to support the claims that the generic omeprazole delayed-release capsules perform differently from the brand omeprazole product in vitro. PMID- 19555237 TI - Hollow spherical nanoparticulate aggregates as potential ultrasound contrast agent: shell thickness characterization. AB - OBJECTIVE: The objective of this work is to manufacture hollow spherical nanoparticulate aggregates for use as an ultrasound contrast agent by means of spray drying of nanoparticulate suspension at a fast drying rate. METHODOLOGY: Biocompatible PMMA-MeOPEGMA and silica nanoparticles are used as the model nanoparticles. The impacts of changing the nanoparticle concentration, pH, and spray drying operating condition on the size and shell thickness-to-particle radius (S/R) ratio, which governs the shell mechanical stability, are investigated. RESULTS AND CONCLUSION: The results indicate that the hollow microspheres size varies between 2 and 10 mum having S/R ratio between 2% and 4%, where the smaller size particles exhibit a higher S/R ratio. The resultant S/R ratio is found to be more influenced by process parameters acting at the nanoparticle scale (e.g., suspension pH) than by the spray drying operating condition. PMID- 19555239 TI - Preparation and characterization of spray-dried mucoadhesive microspheres of aceclofenac. AB - OBJECTIVE: Microencapsulation of the anti-inflammatory drug aceclofenac (ACE) was investigated as a means of controlling drug release and minimizing or eliminating local side effects. METHOD: Microspheres were prepared by a spray-drying technique using solutions of ACE and three polymers, namely, carbopol, chitosan, and polycarbophil, in different weight ratios. RESULTS: The spray-dried mucoadhesive microspheres were characterized in terms of shape (scanning electron microscope), size (6.60-8.40 mum), production yield (34.10-55.62%), and encapsulation efficiency (58.14-90.57%). In vitro release studies were performed in phosphate buffer (pH 6.8) up to 10 hours. The spray-drying process of solutions of ACE with polymeric blends can give prolonged drug release. The in vitro release data were well fit into Higuchi and Korsmeyer-Peppas model and followed Fickian diffusion mechanism. In vivo data showed that the administration of ACE in polymeric microspheres prevented the gastric side effects. CONCLUSION: The formulations here described can be proposed for the oral administration of nonsteroidal anti-inflammatory drugs with minimal side effects on gastric mucosa. PMID- 19555241 TI - Particle size determination of sunscreens formulated with various forms of titanium dioxide. AB - BACKGROUND: There has been some apprehension expressed in the scientific literature that nanometer-sized titanium dioxide (TiO(2)) and other nanoparticles, if able to penetrate the skin, may cause cytotoxicity. In light of a lack of data regarding dermal penetration of titanium dioxide from sunscreen formulations, the Food and Drug Administration Center for Drug Evaluation and Research initiated a study in collaboration with the National Center for Toxicology Research using minipigs to determine whether nanoscale TiO(2) in sunscreen products can penetrate intact skin. Four sunscreen products were manufactured. METHOD: The particle size distribution of three TiO(2) raw materials, a sunscreen blank (no TiO(2)) and three sunscreen formulations containing uncoated nanometer-sized TiO(2), coated nanometer-sized TiO(2) or sub micron TiO(2) were analyzed using scanning electron microscopy (SEM), laser scanning confocal microscopy (LSCM), and X-ray diffraction (XRD) to determine whether the formulation process caused a change in the size distributions (e.g., agglomeration or deagglomeration) of the TiO(2). RESULTS: SEM and XRD of the formulated sunscreens containing nanometer TiO(2) show the TiO(2) particles to have the same size as that observed for the raw materials. This suggests that the formulation process did not affect the size or shape of the TiO(2) particles. CONCLUSION: Because of the resolution limit of optical microscopy, nanoparticles could not be accurately sized using LSCM, which allows for detection but not sizing of the particles. LSCM allows observation of dispersion profiles throughout the sample; therefore, LSCM can be used to verify that results observed from SEM experiments are not solely surface effects. PMID- 19555240 TI - Damar Batu as a novel matrix former for the transdermal drug delivery: in vitro evaluation. AB - PURPOSE: Damar Batu (DB) is a novel film-forming biomaterial obtained from Shorea species, evaluated in this study for its potential application in transdermal drug delivery system. METHODS: DB was characterized initially in terms of acid value, softening point, molecular weight (M(w)), polydispersity index (M(w)/M(n)), and glass transition temperature (T(g)). Neat, plasticized films of DB were investigated for mechanical properties. The biomaterial was further investigated as a matrix-forming agent for transdermal drug delivery system. Developed matrix-type transdermal patches were evaluated for thickness and weight uniformity, folding endurance, drug content, in vitro drug release study, and skin permeation study. RESULTS: On the basis of in vitro drug release and in vitro skin permeation performance, formulation containing DB/Eudragit RL100 (60 : 40) was found to be better than other formulations and was selected as the optimized formulation. IR analysis of physical mixture of drug and polymer and thin layer chromatography study exhibited compatibility between drug and polymer. CONCLUSION: From the outcome of this study, it can be concluded that applying suitable adhesive layer and backing membrane-developed DB/ERL100, transdermal patches can be of potential therapeutic use. PMID- 19555243 TI - Influence of soluble and insoluble cyclodextrin polymers on drug release from hydroxypropyl methylcellulose tablets. AB - BACKGROUND: The influence of beta-cyclodextrin (beta-CD) polymers on drug release from hydroxypropyl methylcellulose (HPMC) matrices has not been reported in the literature. AIM: The influence of monomeric beta-CD and both soluble and insoluble beta-CD polymers on drug release from tablets containing either 30% or 50% hydroxypropyl methylcellulose has been studied using diflunisal (DF) as model drug. METHOD: The DF-beta-CD inclusion complex (1:1 M) was prepared by coevaporation and characterised using X-ray diffraction, differential thermal analysis, and IR spectroscopy. The dissolution assays were performed according to the USP paddle method. RESULTS: The incorporation of beta-CD in the complexed form increases drug release from hydroxypropyl methylcellulose tablets in comparison with the physical mixture because of the better solubilization of the drug. The soluble polymer promotes drug release to a higher extent than the physical mixture with monomeric beta-CD, but the insoluble polymer, which is itself a hydrogel, gives rise to the most retarded release profile, probably by retention of the drug in its structure. The formulations containing physical mixtures with either beta-CD or the soluble polymer present an optimum adjustment to zero-order release kinetics, and the inclusion complex followed non-Fickian diffusion according to the Korsmeyer-Peppas model. CONCLUSION: The release profile of DF from a HPMC matrix can be modulated in different ways by the use of either monomeric or polymeric beta-CD. PMID- 19555242 TI - Comparison of low-shear and high-shear granulation processes: effect on implantable calcium phosphate granule properties. AB - BACKGROUND: Calcium phosphate porous ceramics present a great interest not only as complex bone defect fillers but also as drug delivery systems. Most of the methods described in the literature to fabricate pellets are based on compaction, casting into spherical molds, or on processes such as liquid immiscibility or foaming. Despite wet granulation is used in a wide range of applications in pharmaceuticals, food, detergents, fertilizers, and minerals, it is not applied in the biomaterial field to produce granules. METHODS: In this study physicochemical and in vitro drug delivery properties of implantable calcium phosphate granules, produced by two wet agglomeration processes, were compared. Pellets obtained by high shear granulation (granulation in a Mi-Pro apparatus) were shown to be more spherical and less friable than granules elaborated by low shear process (granulation in a Kenwood apparatus). Although Mi-Pro pellets had a slightly lower porosity compared to Kenwood granules, ibuprofen loading efficiency and dissolution profiles were not statistically different and the release mechanism was mainly controlled by diffusion, in both cases. CONCLUSION: Mi-Pro pellets appeared to be better candidates as bone defect fillers and local drug delivery systems as far as they were more spherical and less friable than Kenwood agglomerates. PMID- 19555244 TI - Transdermal delivery of carvedilol in rats: probing the percutaneous permeation enhancement mechanism of soybean extract-chitosan mixture. AB - BACKGROUND: This study was designed for investigating the effect of soybean (SS) extract and chitosan (CTN) in facilitating the permeation of carvedilol (CDL) across rat epidermis. METHOD: Transdermal flux of carvedilol through heat separated rat epidermis was investigated in vitro using vertical Keshary-Chien diffusion cells. Biophysical and microscopic manifestations of epidermis treated with SS-extract, CTN, and SS extract-CTN mixture were investigated by using DSC, TEWL, SEM, and TEM. Biochemical estimations of cholesterol, sphingosine, and triglycerides were carried out for treated excised as well as viable rat epidermis. The antihypertensive activity of the patches in comparison to that after oral administration of carvedilol was studied in deoxycorticosterone acetate-induced hypertensive rats. RESULTS: The solubility of CDL was found to be maximum in the presence of 1% (w/v) SS extract. The K(IPM/PB) of CDL decreased with increase in concentration of SS extract. The in vitro permeation of CDL across rat epidermis increased and was maximum with combination of SS extract and chitosan (CTN). Biochemical and microscopic studies revealed the initiation of reversal of barrier integrity after 12 hours. Furthermore, the application of patches containing SS extract-CTN mixture resulted in sustained release of carvedilol, which was able to control the hypertension in deoxycorticosterone acetate (DOCA) induced hypertensive rats through 24 hours. CTN was found to potentiate the permeation enhancing activity of SS extract. CONCLUSION: The developed transdermal patches of CDL containing SS extract-CTN mixture exhibited better performance as compared to oral administration in controlling hypertension in rats. PMID- 19555245 TI - Dissolution rate enhancement of the poorly water-soluble drug Tibolone using PVP, SiO2, and their nanocomposites as appropriate drug carriers. AB - BACKGROUND: Creation of immediate release formulations for the poorly water soluble drug Tibolone through the use of solid dispersions (SDs). AIM: SD systems of Tibolone (Tibo) with poly(vinylpyrrolidone) (PVP), fumed SiO(2) nanoparticles, and their corresponding ternary systems (PVP/SiO(2)/Tibo) were prepared and studied in order to produce formulations with enhanced drug dissolution rates. METHOD: The prepared SDs were characterized by the use of differential scanning calorimetry and wide-angle X-ray diffractometry techniques. Also dissolution experiments were performed. RESULTS: From the results it was concluded that PVP as well as SiO(2) can be used as appropriate carriers for the amorphization of Tibo, even when the drug is used at high concentrations (20-30%, w/w). This is due to the evolved interactions taking place between the drug and the used carriers, as was verified by Fourier transform infrared spectroscopy. At higher concentrations the drug was recrystallized. Similar are the observations on the ternary PVP/SiO(2)/Tibo SDs. The dissolution profiles of the drug in PVP/Tibo and SiO(2)/Tibo SDs are directly dependent on the physical state of the drug. Immediately release rates are observed in SD with low drug concentrations, in which Tibo was in amorphous state. However, these release profiles are drastically changed in the ternary PVP/SiO(2)/Tibo SDs. An immediate release profile is observed for low drug concentrations and an almost sustained release as the concentration of Tibo increases. This is due to the weak interactions that take place between PVP and SiO(2), which result in alterations of the characteristics of the carrier (PVP/SiO(2) nanocomposites). CONCLUSIONS: Immediate release formulation was created for Tibolone as well as new nanocomposite matrices of PVP/SiO((2)), which drastically change the release profile of the drug to a sustained delivery. PMID- 19555246 TI - Spray-drying process optimization for manufacture of drug-cyclodextrin complex powder using design of experiments. AB - BACKGROUND: Design of experiments (DOE), a component of Quality by Design, is systematic and simultaneous evaluation of variables (process or formulation) to develop a product with predetermined quality attributes. This study presents a case study to understand the effects of process variables in a spray-drying process used in the manufacture of drug-cyclodextrin complex for a drug that is prone to chemical instability at elevated temperature conditions encountered during processing. METHODS: Experiments were designed, and data were collected according to a three-factor, three-level face-centered central composite design. The factors investigated were inlet temperature, spray rate, and batch size. Responses analyzed for computing the interaction effects were drug content, impurities, moisture content, and process yield. The spray-drying process conditions were optimized using DOE to maximize production yields while minimizing moisture content and drug-related impurities. Process validation batches were executed using the optimum process conditions obtained from software Design-Expert((R)) to evaluate both the repeatability and reproducibility of spray-drying technique. RESULTS: Optimization of process variables using DOE resulted in a significant improvement of process yields, above 90% and moisture content below 6% (w/w). The impurities were controlled within acceptable limits. The desirability function used to optimize the response variables and observed responses were in agreement with experimental values. These results demonstrated the reliability of selected model for manufacture of powder complex with predictable quality attributes. CONCLUSION: The study indicates the general applicability of DOE approach to optimize critical process parameters in the manufacture of drug product with desired quality attributes. PMID- 19555247 TI - Effect of unconventional curing conditions and storage on pellets coated with Aquacoat ECD. AB - PURPOSE: Purpose of this study was to develop storage stable pellets coated with the aqueous ethylcellulose dispersion Aquacoat ECD. METHODS: The influence of accelerated curing/storage conditions on the release behavior of Aquacoat/HPMC coated drug pellets were investigated as a function of various formulations (sealing, plasticizer content, and pore-former type/amount) and process parameters (process humidity, thermal curing, and organic processing). RESULTS: Conventionally cured Aquacoat/hydroxypropyl methylcellulose- coated pellets were storage stable at ambient conditions and 25 degrees C/60% relative humidity (RH) but showed a decreasing drug release at 40 degrees C/75% RH, which is a required test condition according to ICH guidelines. CONCLUSION: Only organic processing of dried Aquacoat or unconventionally harsh curing conditions (60 degrees C/75% RH or 80 degrees C) improved the storage stability of Aquacoat-coated pellets at accelerated conditions. PMID- 19555248 TI - The influence of porosity changes in human epidermal membrane during iontophoresis on the permeability enhancement of a model peptide. AB - PURPOSE: We tested the hypothesis that the increases in the porosity of the skin during iontophoresis would not significantly increase the transport of peptides due to the small size of electrically induced pores. To investigate this mechanistically, we used human epidermal membrane under constant voltage conditions, applying the Nernst-Planck equation to the transport of a small ionic solute, tetraethylammonium bromide (TEAB), and a model peptide, luteinizing hormone releasing hormone. METHODS: Steady-state flux of the drugs was determined under passive conditions and also during iontophoresis using constant DC voltages applied across side-by-side diffusion cells. Electrical conductance measurements were used to monitor the porosity changes that occur during electrical field application. RESULTS: Porosity increases observed in the membrane substantially increased the permeability enhancement of the small ionic solute TEAB. The permeability enhancement was well described by Nernst-Planck model predictions after porosity changes in the membrane were taken into account. Enhancement of luteinizing hormone releasing hormone under identical conditions was much less than TEAB. The porosity increases induced by iontophoresis had little or no effect on the permeability enhancement of the larger molecule. CONCLUSIONS: These findings closely parallel those reports that have found electrically induced pores to be significantly smaller than preexisting pores in the human epidermal membrane. The data obtained also support the view that iontophoresis-induced pores, alone, may provide limited benefit for macromolecule transport across the skin. PMID- 19555250 TI - Citric acid as a pH-modifying additive in an extended release pellet formulation containing a weakly basic drug. AB - BACKGROUND: An extended release pellet formulation (ACES(R)) of the weakly basic drug propiverine was developed with spheronized citric acid crystals as starter cores. METHOD: Coated pellets, consisting of several layers of functional coatings, were manufactured by fluid bed coating. Different coating levels were examined with regard to their effect on drug release. Release profiles from the formulations with or without pH modifier and the free base as well as the hydrochloride salt of the active ingredient were compared. RESULTS: The coated citric acid starter cores led to a controlled release of the drug and the pH modifier, resulting from modulation of the microenvironmental pH throughout the dissolution period of 17 hours. If microcrystalline cellulose pellets are used as starter cores drug release is strongly pH-dependent. Significant differences in the drug release profiles were observed between the formulations containing the free drug base and those with the hydrochloride salt as a result of an altered microenvironmental pH. CONCLUSION: The presented extended release pellet formulation is able to maintain a low pH within the pellet core and thus a sufficiently high drug solubility. By maintaining a low pH inside the pellets, a controlled drug release can be achieved. PMID- 19555249 TI - Processing difficulties and instability of carbohydrate microneedle arrays. AB - BACKGROUND: A number of reports have suggested that many of the problems currently associated with the use of microneedle (MN) arrays for transdermal drug delivery could be addressed by using drug-loaded MN arrays prepared by moulding hot melts of carbohydrate materials. METHODS: In this study, we explored the processing, handling, and storage of MN arrays prepared from galactose with a view to clinical application. RESULTS: Galactose required a high processing temperature (160 degrees C), and molten galactose was difficult to work with. Substantial losses of the model drugs 5-aminolevulinic acid (ALA) and bovine serum albumin were incurred during processing. While relatively small forces caused significant reductions in MN height when applied to an aluminium block, this was not observed during their relatively facile insertion into heat-stripped epidermis. Drug release experiments using ALA-loaded MN arrays revealed that less than 0.05% of the total drug loading was released across a model silicone membrane. Similarly, only low amounts of ALA (approximately 0.13%) and undetectable amounts of bovine serum albumin were delivered when galactose arrays were combined with aqueous vehicles. Microscopic inspection of the membrane following release studies revealed that no holes could be observed in the membrane, indicating that the partially dissolved galactose sealed the MN-induced holes, thus limiting drug delivery. Indeed, depth penetration studies into excised porcine skin revealed that there was no significant increase in ALA delivery using galactose MN arrays, compared to control (P value < 0.05). Galactose MNs were unstable at ambient relative humidities and became adhesive. CONCLUSION: The processing difficulties and instability encountered in this study are likely to preclude successful clinical application of carbohydrate MNs. The findings of this study are of particular importance to those in the pharmaceutical industry involved in the design and formulation of transdermal drug delivery systems based on dissolving MN arrays. It is hoped that we have illustrated conclusively the difficulties inherent in the processing and storage of carbohydrate-based dissolving MNs and that those in the industry will now follow alternative approaches. PMID- 19555251 TI - Characterization of ethylcellulose: starch-based film coatings for colon targeting. AB - BACKGROUND: The site-specific delivery of drugs to the colon can be highly advantageous for various applications, including the local treatment of inflammatory bowel diseases. The aim of this study was to provide efficient tools that can be used to easily adjust the key properties of novel polymeric film coatings allowing for colon targeting. METHODS: Free films based on blends of ethylcellulose and different types of starch derivatives (partially being pregelatinized, acetylated, and/or hydroxypropylated) were prepared and characterized. RESULTS: The key properties of the polymeric systems can effectively be adjusted by varying the polymer blend ratio and type of starch derivative. This includes the water uptake and dry mass loss kinetics as well as the mechanical properties of the films before and upon exposure to aqueous media simulating the contents of the upper GIT. CONCLUSION: Broad ranges of film coating properties can easily be provided, being adapted to the needs of the respective drug treatment. PMID- 19555252 TI - Phospholipid-based ultrasonic microbubbles for loading protein and ultrasound triggered release. AB - BACKGROUND: Ultrasonic microbubbles are used as ultrasound-triggered delivery carriers for protein drugs. AIM: This work was to prepare stabilized protein loaded phospholipid-based ultrasonic microbubbles (PUM) and to determine its value as a protein delivery system. METHOD: Bovine serum albumin (BSA) was used as a model protein drug. BSA-containing PUM were prepared by dissolving lyophilized PUM powder in BSA solution. The particle size and microbubble concentration of BSA-containing PUM were measured. The BSA encapsulation efficiency as a function of BSA concentration was determined. Contrast enhancement of BSA-containing PUM in vivo was detected. The release profile of BSA from PUM was also investigated. RESULTS: The mean particle size and microbubble concentration of PUM were unchanged by the presence of BSA for at least 30 minutes after preparation. The net amount of BSA entrapped in PUM was maintained unchanged with increasing BSA concentration. BSA-containing PUM were shown easily to be visible in in vivo rabbit kidney. There was no difference in echogenicity between the loaded and unloaded PUM. Ultrasound duration had a positive relationship with BSA release. Ultrasound of 30 seconds stimulated 94.1% and 93.3% of BSA release from PUM solutions containing 0.3% and 1.5% BSA, respectively. CONCLUSIONS: Protein-loaded PUM exhibited satisfactory physical characteristics and were potent for using in ultrasound-triggered delivery. PMID- 19555253 TI - Fenofibrate, simvastatin and their combination in the management of dyslipidaemia in type 2 diabetic patients. AB - OBJECTIVE: To evaluate the efficacy of fenofibrate, simvastatin or their combination in type 2 diabetic patients with combined dyslipidaemia. RESEARCH DESIGN AND METHODS: 241 patients, who had never previously taken lipid-lowering medications, received fenofibrate 145 mg/day, or simvastatin 40 mg/day, or fenofibrate 145 mg/day + simvastatin 40 mg/day combination for 12 months. We evaluated lipids, glycaemic, haemostatic, and inflammatory variables at baseline, and after 6 and 12 months. RESULTS: After 12 months total cholesterol (TC), LDL cholesterol (LDL-C) and triglycerides (Tg) decreased while HDL cholesterol (HDL C) increased in all groups, even if the values obtained with fenofibrate + simvastatin were the best. At the end of the study apolipoprotein A-1 (Apo A-1) increased with fenofibrate + simvastatin, while apolipoprotein B (Apo B) decreased in all groups compared to baseline. Plasminogen activator inhibitor-1 (PAI-1) and high-sensitivity C reactive protein (hs-CRP) decreased after 12 months compared to baseline with simvastatin, and with fenofibrate + simvastatin even if the value obtained with fenofibrate-simvastatin was the lowest. After 12 months, fibrinogen (Fg) decreased compared to baseline with fenofibrate + simvastatin. LIMITATIONS: This study has some limitations. The first one is the relatively small sample of studied patients. The second one is the lack of an advanced lipid proteins evaluation, such as lipoprotein subfraction changes in the different treatment regimen. Finally, we have not selected patients that could show the best response to fibrate (i.e.: hypertriglyceridemics) or statins (i.e.: hypercholesterolemics) monotherapy, so the effect of these drugs administered alone may have been partly attenuated. CONCLUSIONS: Fenofibrate + simvastatin association improved lipid parameters, prothrombotic and inflammatory factors, and appeared to have a good tolerability profile over 12 months of therapy. PMID- 19555265 TI - Colonic treatments and targets: issues and opportunities. AB - The colon provides a plethora of therapeutic opportunities. There are multiple disease targets, drug molecules, and colon-specific delivery systems to be explored. Clinical studies highlight the potential for systemic delivery via the colon, and the emerging data on the levels of cell membrane transporters and metabolic enzymes along the gut could prove advantageous for this. Often efflux transporters and metabolic enzyme levels are lower in the colon, suggesting a potential for improved bioavailability of drug substrates at this site. The locoregional distribution of multiple metabolic enzymes (including cytochromes), efflux transporters (including P-glycoprotein and breast cancer resistance proteins), and influx transporters (including the solute carrier family) along the intestine is summarized. Local delivery to the colonic mucosa remains a valuable therapeutic option. New therapies that target inflammatory mediators could improve the treatment of inflammatory bowel disease, and old and new anticancer molecules could, when delivered topically, prove to be beneficial adjuncts to the current systemic or surgical treatments. New issues such as pharmacogenomics, chronotherapeutics, and the delivery of prebiotics and probiotics are also discussed in this review. Targeting drugs to the colon utilizes various strategies, each with their advantages and flaws. The most promising systems are considered in the light of the physiological data which influence their in vivo behavior. PMID- 19555266 TI - Controlled release of dexamethasone acetate from biodegradable and biocompatible polyurethane and polyurethane nanocomposite. AB - Polyurethanes and polyurethane nanocomposites can be applied to control the release of drugs previously incorporated into these materials. In this study, dexamethasone acetate (ACT) was incorporated into biodegradable and biocompatible polyurethane and polyurethane containing montmorillonite nanoparticles. Fourier transform infrared spectroscopic technique showed no strong interactions between drug and polymers. Data obtained from X-ray diffraction and small angle X-ray scattering indicated that the incorporation of ACT did not disturb the polymer morphology, but montmorillonite led to a less defined phase separation between hard and soft segments of polyurethane. The in vitro release studies demonstrated that nanoparticles increased the rate of ACT release possibly because these particles have a hydrophilic surface that increases the absorption of water and accelerates the hydrolysis of the polymer. The in vivo short-term biocompatibility studies demonstrated adequate interfacial interaction between polyurethane and subcutaneous tissue and a discreet inflammatory response which was completely resolved in 14 days. PMID- 19555267 TI - Preparation, characterization, and biodistribution of breviscapine proliposomes in heart. AB - Breviscapine proliposomes were prepared by ethanol injection-homogenization lyophilization method. On contact with 5% glucose, the proliposomes were rapidly converted into a liposomal dispersion, in which a certain amount of breviscapine was entrapped by the liposomes. The entrapment efficiency measured by reverse dialysis method was 77.89 +/- 0.28%. The particle size, polydispersity index, and zeta potential of breviscapine liposomes were 504.83 +/- 52.88 nm (by intensity), 0.17 +/- 0.02, and -(20.31 +/- 1.03) mV, respectively (mean +/- SD, n = 3). In mimic-biomembrane model experiment, breviscapine was distributed not only to n octanol and buffer phase but also to interfacial phase. After bolus administration, the elimination phase (t(1/2(beta)) = 66.386) of liposomal formulation in plasma was 4.8 times longer than that of solution formulation (t(1/2(beta)) = 13.695). The AUC and MRT values of liposomal formulation in heart were increased more than 11.7- and 3.2-fold versus solution formulation, respectively. These results were all beneficial to heart disease therapy. PMID- 19555268 TI - Compulsory licensing and the AIDS epidemic in South Africa. AB - One cannot look at the high prevalence and persistent, dramatic spread of HIV in developing countries, especially those in Sub-Saharan Africa, without also examining economic and public health systems. Recent developments in South Africa have highlighted an emerging controversy surrounding the issue of access to essential medicines and drug pricing. Sparked by a South African amendment, Article 15(C) of the Medicines and Related Substances Act 101, passed in 1997, that would allow the South African Minister of Health to provide less expensive generic forms of essential medicines, including anti-HIV therapies, to South Africa, law- and policymakers as well as activists have been taking sides in a complicated debate that mixes important health issues with international trade law, patent protection, economic incentives, and United States domestic law. Threatened by what could be a weakening of patent protection, pharmaceutical firms in the United States, South Africa, and Europe have generally opposed the efforts of the South African government to allow production of such generic products, claiming that the 1997 amendment violates international trade law, particularly the Trade-Related Aspects of Intellectual Property Rights (TRIPS) Agreement. AIDS activists and consumer advocates generally support the efforts of the South African government, claiming that international law is flexible in cases of national emergency. Lobbied by the pharmaceutical industry, United States government officials have proposed bilateral trade sanctions in an effort to pressure South Africa to repeal the 1997 amendment. These issues are reviewed in this report. Relevant provisions of Article 15(C) and the TRIPS Agreement are available on the Internet or from the American Foundation for AIDS Research (amfAR). PMID- 19555269 TI - Nocardiasis in patients with HIV infection. PMID- 19555271 TI - Antiretroviral therapy of pediatric HIV infection: making hope a reality. AB - Over the past 3 years, the treatment and prognosis of HIV-1 infection have been revolutionized by a better understanding of the pathogenesis of HIV-1 infection, the ability to monitor viral replication and drug resistance in the host, and the availability of potent combination chemotherapy. While most of the studies that have led to this transformation have been done in adults, the results can be applied to the care of children. Data from trials of highly active antiretroviral therapy (HAART) in children are now being presented or published. Although the basic principles of antiretroviral therapy of HIV-1 infection do not differ between adults and children, there are important differences in the natural history of the disease and in issues related to medication administration and adherence to therapy. Progression of disease may be more rapid in children and is often very rapid in infants. Administration of medication to infants and children can be difficult, especially when the medication tastes bad. Finally, whereas an adult patient is free to decline therapy, however foolish such a decision may seem to the health-care professional, the failure to administer effective medication to a child for a condition that threatens serious morbidity or death constitutes medical neglect. In this review we will discuss the basic principles underlying pediatric antiretroviral therapy and address the issue of adherence, the major impediment to treatment success. PMID- 19555272 TI - Pelvic inflammatory disease in adolescent girls. PMID- 19555273 TI - Coping of staff in AIDS nursing facilities. AB - The primary objective of this study was to examine coping among nursing staff who work in nursing facilities for AIDS patients. The study explored the relationships between nurses' personal coping resources, availability and use of worksite support services, and nurses' attitudes toward dying AIDS patients. Self administered questionnaires were distributed to 437 nursing staff in seven AIDS residential health-care facilities in New York City. A total of 197 completed questionnaires were returned (48% response rate). Measures of demographics, attitudes toward caring for dying patients, personal coping resources, and worksite support were included. There was a significant difference between different levels of nursing staff in attitudes toward caring for dying patients, with certified nursing assistants reporting more negative attitudes than licensed practical nurses and registered nurses. Staff with advanced degrees reported more personal coping resources and more positive attitudes toward caring for dying patients than those with lower education. Participants who reported more social, physical, and spiritual coping resources also reported more positive attitudes toward caring for dying patients. Worksite support services should be offered to all workers in AIDS nursing facilities, and services should be tailored to meet the needs of lower level nursing staff and those with less education who report fewer coping resources and less positive attitudes toward patients. PMID- 19555274 TI - Ethnic, demographic, and social differences among middle and older adults with HIV/AIDS. AB - Although the prevalence of AIDS among middle and older adults is increasing, little is known about them. The objective of this study was to obtain basic demographic and social information about people with HIV/AIDS (PWAs) over the age of 44 years. This was a descriptive, cross-sectional survey of convenience samples of 76 African-American (AA) and 80 White older PWAs. Participants were interviewed by trained peer interviewers using a structured and largely objective schedule. Participants and interviewers were each paid $35 per completed interview. The schedule consisted of wide-ranging demographic, HIV/AIDS, stressor, coping, social, and support questions. Although both ethnic groups had incomes significantly below national norms and poverty rates three times their national rates, AA PWAs were markedly disadvanted socioeconomically relative to White PWAs. Although over 60% of all PWAs reported that having enough money to live on was a problem, AAs also reported significantly more Stressors, many related to economics, then White PWAs. AA PWAs reported being predominantly heterosexual, while the largest group of Whites was homosexual or bisexual. Sources of infection paralleled sexual orientation. Both groups used relatively effective coping strategies and reported moderate levels of social support and activity. It was concluded that the course of illness, treatment, and quality of life of middle and older PWAs are likely to be complicated by economic factors, and this will be particularly serious among AA PWAs. PMID- 19555275 TI - Strongyloidiasis detected by the agar plate culture method among patients infected by HIV. AB - To evaluate the rate of strongyloidiasis among HIV/AIDS patients, stools and duodenal juice were examined using the agar plate culture method. From January to June 1993, a total of 60 HIV/AIDS patients were required for duodenal aspirate and three serial samples of freshly passed stools. Stools and duodenal aspirate were dispensed on an agar plate culture; after incubation at 28 degrees C during 48 h, screening of plates was made at 10 x. The presence of furrows and worms of short buccal chamber and prominent genital primordium were positive for Strongyloides stercoralis. Most patients were men (91.7%); their mean age, of 33.9 years (SD +/- 10.6); their median CD4(+) T-cells count, of 105/microL (range of 12 to 646). S. stercoralis was detected in three patients (5%). In duodenal juice, the three patients showed the parasite, but in feces, only two (3.3%). In these two individuals, the worms were found in feces by agar culture and Faust's concentration method. The rate of S. stercoralis in feces of HIV/AIDS individuals (3.3%) by agar culture method was similar to that formerly reported from the general Mexican population (2.9%) using standard concentration procedures. Hence, in this immunocompromised population of a low prevalence city, there was no advantage to using an agar plate culture for strongyloidiasis. PMID- 19555276 TI - Antiviral briefs. PMID- 19555277 TI - Drug developments and STD news. PMID- 19555284 TI - Antimicrobial agents for complicated skin and skin-structure infections: noninferiority margins, placebo-controlled trials, and the complexity of clinical trials. PMID- 19555285 TI - Antimicrobial agents for complicated skin and skin-structure infections: justification of noninferiority margins in the absence of placebo-controlled trials. AB - BACKGROUND: The United States Food and Drug Administration requires clinical trial noninferiority margins to preserve a fraction (eg, 50%) of the established comparator drug's efficacy versus placebo. Lack of placebo-controlled trials for many infections complicates noninferiority margin justification for and, hence, regulatory review of new antimicrobial agents. Noninferiority margin clarification is critical to enable new antimicrobial development. In the absence of placebo-controlled trials, we sought to define the magnitude of efficacy of antimicrobial agents and resulting noninferiority margins for studies of complicated skin and skin-structure infection (SSSI). METHODS: We systematically reviewed literature on complicated SSSI published during 1900-1950 (before widespread penicillin resistance) to define treatment outcomes and confidence intervals (CIs). Antimicrobial efficacy was calculated as the lower limit CI of the cure rate with antimicrobials minus the upper limit CI of the cure rate without antimicrobials. RESULTS: We identified 90 articles describing >28,000 patients with complicated SSSI. For cellulitis/erysipelas, cure rates were 66% (95% CI, 64%-68%) without antibiotics and 98% (95% CI, 96%-99%) for penicillin treated patients, and penicillin reduced mortality by 10%. Cure rates for wound/ulcer infections were 36% (95% CI, 32%-39%) without antibiotics and 83% (95% CI, 81%-85%) for penicillin-treated patients. For major abscesses, cure rates were 76% (95% CI, 71%-80%) without antibiotics and 96% (95% CI, 94%-98%) for penicillin-treated patients; penicillin reduced mortality by 6%. CONCLUSION: Systematic review of historical literature enables rational noninferiority margin justification in the absence of placebo-controlled trials and may facilitate regulatory review of noninferiority trials. Noninferiority margins of 14% for cellulitis/erysipelas, 21% for wound/ulcer infections, and 7% for major abscesses would preserve >or= 50% of antibiotic efficacy versus placebo for these complicated SSSI subsets. PMID- 19555286 TI - A prediction rule for estimating the risk of bacteremia in patients with community-acquired pneumonia. AB - BACKGROUND: We endeavored to construct a simple score based entirely on epidemiological and clinical variables that would stratify patients who require hospital admission because of community-acquired pneumonia into groups with a low or high risk of developing bacteremia. METHODS: Derivation and internal validation cohorts were obtained by retrospective analysis of a database that included 3116 consecutive patients with community-acquired pneumonia from 2 university hospitals. Potential predictive factors were determined by means of a multivariate logistic regression equation applied to a cohort consisting of 60% of the patients. Points were assigned to significant parameters to generate the score. It was then internally validated with the remaining 40% of patients and was externally validated using an independent multicenter cohort of 1369 patients. RESULTS: The overall rates of bacteremia were 12%-16% in the cohorts. The clinical probability estimate of developing bacteremia was based on 6 variables: liver disease, pleuritic pain, tachycardia, tachypnea, systolic hypotension, and absence of prior antibiotic treatment. For the score, 1 point was assigned to each predictive factor. In the derivation cohort, a cutoff score of 2 best identified the risk of bacteremia. In the validation cohorts, rates of bacteremia were <8% for patients with a score 1 (43%-49% of patients), whereas blood culture results were positive in 14%-63% of cases for patients with a score 2. CONCLUSIONS: This clinical score, based on readily available and objective variables, provides a useful tool to predict bacteremia. The score has been internally and externally validated and may be useful to guide diagnostic decisions for community-acquired pneumonia. PMID- 19555287 TI - Primary pneumonic plague contracted from a mountain lion carcass. AB - BACKGROUND: Primary pneumonic plague is a rare but often fatal form of Yersinia pestis infection that results from direct inhalation of bacteria and is potentially transmissible from person to person. We describe a case of primary pneumonic plague in a wildlife biologist who was found deceased in his residence 1 week after conducting a necropsy on a mountain lion. METHODS: To determine cause of death, a postmortem examination was conducted, and friends and colleagues were interviewed. Physical evidence was reviewed, including specimens from the mountain lion and the biologist's medical chart, camera, and computer. Human and animal tissues were submitted for testing. Persons in close contact (within 2 meters) to the biologist after he had developed symptoms were identified and offered chemoprophylaxis. RESULTS: The biologist conducted the necropsy in his garage without the use of personal protective equipment. Three days later, he developed fever and hemoptysis and died approximately 6 days after exposure. Gross examination showed consolidation and hemorrhagic fluid in the lungs; no buboes were noted. Plague was diagnosed presumptively by polymerase chain reaction and confirmed by culture. Tissues from the mountain lion tested positive for Y. pestis, and isolates from the biologist and mountain lion were indistinguishable by pulsed-field gel electrophoresis. Among 49 contacts who received chemoprophylaxis, none developed symptoms consistent with plague. CONCLUSIONS: The biologist likely acquired pneumonic plague through inhalation of aerosols generated during postmortem examination of an infected mountain lion. Enhanced awareness of zoonotic diseases and appropriate use of personal protective equipment are needed for biologists and others who handle wildlife. PMID- 19555288 TI - Treatment responses in antiretroviral treatment-naive premenopausal and postmenopausal HIV-1-infected women: an analysis from AIDS Clinical Trials Group Studies. AB - Menopause may affect antiretroviral treatment (ART) response. Immunologic and virologic responses to ART were compared in 220 premenopausal and 47 postmenopausal women enrolled in 2 studies involving ART-naive persons. Changes in CD4 counts or human immunodeficiency virus type 1 RNA levels were similar at 24, 48, and 96 weeks after treatment initiation. ART-naive women should respond to ART regardless of menopausal status. PMID- 19555289 TI - The glial nature of embryonic and adult neural stem cells. AB - Glial cells were long considered end products of neural differentiation, specialized supportive cells with an origin very different from that of neurons. New studies have shown that some glial cells--radial glia (RG) in development and specific subpopulations of astrocytes in adult mammals--function as primary progenitors or neural stem cells (NSCs). This is a fundamental departure from classical views separating neuronal and glial lineages early in development. Direct visualization of the behavior of NSCs and lineage-tracing studies reveal how neuronal lineages emerge. In development and in the adult brain, many neurons and glial cells are not the direct progeny of NSCs, but instead originate from transit amplifying, or intermediate, progenitor cells (IPCs). Within NSCs and IPCs, genetic programs unfold for generating the extraordinary diversity of cell types in the central nervous system. The timing in development and location of NSCs, a property tightly linked to their neuroepithelial origin, appear to be the key determinants of the types of neurons generated. Identification of NSCs and IPCs is critical to understand brain development and adult neurogenesis and to develop new strategies for brain repair. PMID- 19555291 TI - Cerebellum and nonmotor function. AB - Does the cerebellum influence nonmotor behavior? Recent anatomical studies demonstrate that the output of the cerebellum targets multiple nonmotor areas in the prefrontal and posterior parietal cortex, as well as the cortical motor areas. The projections to different cortical areas originate from distinct output channels within the cerebellar nuclei. The cerebral cortical area that is the main target of each output channel is a major source of input to the channel. Thus, a closed-loop circuit represents the major architectural unit of cerebro cerebellar interactions. The outputs of these loops provide the cerebellum with the anatomical substrate to influence the control of movement and cognition. Neuroimaging and neuropsychological data supply compelling support for this view. The range of tasks associated with cerebellar activation is remarkable and includes tasks designed to assess attention, executive control, language, working memory, learning, pain, emotion, and addiction. These data, along with the revelations about cerebro-cerebellar circuitry, provide a new framework for exploring the contribution of the cerebellum to diverse aspects of behavior. PMID- 19555290 TI - The neuropsychopharmacology of fronto-executive function: monoaminergic modulation. AB - We review the modulatory effects of the catecholamine neurotransmitters noradrenaline and dopamine on prefrontal cortical function. The effects of pharmacologic manipulations of these systems, sometimes in comparison with the indoleamine serotonin (5-HT), on performance on a variety of tasks that tap working memory, attentional-set formation and shifting, reversal learning, and response inhibition are compared in rodents, nonhuman primates, and humans using, in a behavioral context, several techniques ranging from microiontophoresis and single-cell electrophysiological recording to pharmacologic functional magnetic resonance imaging. Dissociable effects of drugs and neurotoxins affecting these monoamine systems suggest new ways of conceptualizing state-dependent fronto executive functions, with implications for understanding the molecular genetic basis of mental illness and its treatment. PMID- 19555296 TI - Air to muscle O2 delivery during exercise at altitude. AB - Hypoxia-induced hyperventilation is critical to improve blood oxygenation, particularly when the arterial Po2 lies in the steep region of the O2 dissociation curve of the hemoglobin (ODC). Hyperventilation increases alveolar Po2 and, by increasing pH, left shifts the ODC, increasing arterial saturation (Sao2) 6 to 12 percentage units. Pulmonary gas exchange (PGE) is efficient at rest and, hence, the alveolar-arterial Po2 difference (Pao2-Pao2) remains close to 0 to 5mm Hg. The (Pao2-Pao2) increases with exercise duration and intensity and the level of hypoxia. During exercise in hypoxia, diffusion limitation explains most of the additional Pao2-Pao2. With altitude, acclimatization exercise (Pao2-Pao2) is reduced, but does not reach the low values observed in high altitude natives, who possess an exceptionally high DLo2. Convective O2 transport depends on arterial O2 content (Cao2), cardiac output (Q), and muscle blood flow (LBF). During whole-body exercise in severe acute hypoxia and in chronic hypoxia, peak Q and LBF are blunted, contributing to the limitation of maximal oxygen uptake (Vo2max). During small-muscle exercise in hypoxia, PGE is less perturbed, Cao2 is higher, and peak Q and LBF achieve values similar to normoxia. Although the Po2 gradient driving O2 diffusion into the muscles is reduced in hypoxia, similar levels of muscle O2 diffusion are observed during small-mass exercise in chronic hypoxia and in normoxia, indicating that humans have a functional reserve in muscle O2 diffusing capacity, which is likely utilized during exercise in hypoxia. In summary, hypoxia reduces Vo2max because it limits O2 diffusion in the lung. PMID- 19555297 TI - Nervous system function during exercise in hypoxia. AB - Aerobic exercise capacity decreases with exposure to hypoxia. This article focuses on the effects of hypoxia on nervous system function and the potential consequences for the exercising human. Emphasis is put on somatosensory muscle afferents due to their crucial role in the reflex inhibition of muscle activation and in cardiorespiratory reflex control during exercise. We review the evidence of hypoxia influences on muscle afferents and discuss important consequences for exercise performance. Efferent (motor) nerves are less affected at altitude and are thought to stay fairly functional even in severe levels of arterial hypoxemia. Altitude also alters autonomic nervous system functions, which are thought to play an important role in the regulation of cardiac output and ventilation. Finally, the consequences of hypoxia-induced cortical adaptations and dysfunctions are evaluated in terms of neurotransmitter turnover, brain electrical activity, and cortical excitability. Even though the cessation of exercise or the reduction of exercise intensity, when reaching maximum performance, implies reduced motor recruitment by the nervous system, the mechanisms that lead to the de-recruitment of active muscle are still not well understood. In moderate hypoxia, muscle afferents appear to play an important role, whereas in severe hypoxia brain oxygenation may play a more important role. PMID- 19555298 TI - Current status of medical training in mountain rescue in America and Europe. AB - Limited medical training of mountain rescuers may adversely affect the outcome of casualties. Thus, this study evaluated medical training of mountain rescuers in countries associated with the International Commission of Mountain Emergency Medicine. A questionnaire was completed by 33 mountain rescue services from 18 countries in America and Europe. First-aid topics taught most often are (absolute values, percentage): chest compression, hypothermia, cold injuries (32 of 33 organization 97%); avalanche rescue, first-aid kit of rescuer, cervical collar (31, 94%); hemorrhagic shock, automated blood pressure measurement, wound dressing (30, 91%); and heat injuries and SAM SPLINT (29, 88%). Cardiopulmonary resuscitation manikins are used in 32 (97%) organizations, and in 17 (52%) organizations manikins have feedback functionality. After training, exams are compulsory in 27 (83%) organizations. Yearly retraining is done in 12 (36%) organizations; 22 (67%) organizations would like to increase medical training. The study shows high variability in the medical training programs among the surveyed organizations and the need to improve medical education. The authors recommend standardization of medical training and examinations on an international level. Additional topics tailored to the typical injury and illness patterns of a particular area should supplement this core training. Training should be performed by highly qualified instructors on a yearly basis. PMID- 19555299 TI - Glioblastoma multiforme: a review of where we have been and where we are going. AB - Malignant gliomas such as glioblastoma multiforme (GBM) present some of the greatest challenges in the management of cancer patients worldwide, despite notable recent achievements in oncology. Even with aggressive surgical resections using state-of-the-art preoperative and intraoperative neuroimaging, along with recent advances in radiotherapy and chemotherapy, the prognosis for GBM patients remains dismal: median survival after diagnosis is about 14 months. Established good prognostic factors are limited, but include young age, high Karnofsky Performance Status (KPS), high mini-mental status examination score, O6 methylguanine methyltransferase promoter methylation, and resection of > 98% of the tumor. Standard treatment includes resection, followed by concurrent chemotherapy and radiotherapy. GBM research is being conducted worldwide at a remarkable pace, with some of the more recent promising studies focused on identification of aberrant genetic events and signaling pathways, tumor stem cell identification and characterization, modulation of tumor immunological responses, combination therapies, and understanding of the rare long-term survivors. Past treatment strategies have failed for various reasons; however, newer strategies in trials today and on the horizon encourage optimism. To help illustrate 'where we have been' with this fatal disease and 'where we are going' with contemporary studies, we include in this review a detailed history of Phase III clinical trials for GBM, with a final emphasis on exciting new treatment strategies that offer hope for future GBM therapy. PMID- 19555302 TI - Preparation, characterization, and in vitro release of new transdermal methimazole as alternative to oral delivery. AB - Our objective has been the development and study of the stability of transdermal methimazole formulae as alternative to oral administration. Preparation of F-127 and PLO Pluronic gel (Pluronic lecitin organogel) are described, as well as their characteristics from transmission electron microscopy. The possible structural and rheological changes to both transdermal forms were studied in terms of composition, temperature and time. The trial period was from 24 hr to 3 months after preparation. Furthermore, identical tests were carried out on formulae conserved for 1 year after production to check their integrity. Studies of release in vitro were carried out showing that the selected excipients do not pose an obstacle to the cession of methimazole, even though the PLO samples were made more slowly. PMID- 19555303 TI - In vitro transdermal iontophoretic delivery of penbutolol sulfate. AB - Iontophoretic transport of penbutolol sulfate across porcine ear skin was studied. Passive transdermal flux of the drug in phosphate-buffered saline was 7.65 microg/cm(2) hr. There was statistically significant flux enhancement when direct current iontophoresis was applied. Iontophoresis (0.11 mA/cm(2), 0.17 mA/cm(2), and 0.22 mA/cm(2)) for 6 hr, resulted in net transport of 87.36 microg/cm(2), 137.51 microg/cm(2), and 201.12 microg/cm(2) of penbutolol sulfate, respectively. After 24 hr, cumulative amount of penbutolol transported were 201.63, 300.76, and 359.98 microg/cm(2), respectively. There was a 2.20- (0.11 mA/cm(2)), 3.26- (0.17 m/Acm(2)), and 4.28-fold (0.22 mA/cm(2)) enhancement in transcutaneous steady-state flux values compared to passive delivery. Steady state fluxes of penbutolol sulfate also increased proportionally to current density. Steady-state fluxes calculated from the linear portion of the cumulative amount versus time curves for penbutolol sulfate were 16.68, 24.97, and 32.76 microg/cm(2)/hr at current densities of 0.11, 0.17, and 0.22 mA/cm(2). This study provides initial evidence for the potential use of iontophoresis for enhanced transdermal delivery of penbutolol sulfate. PMID- 19555300 TI - Can we predict the effects of NF-kappaB inhibition in sepsis? Studies with parthenolide and ethyl pyruvate. AB - BACKGROUND: Based partially on encouraging findings from preclinical models, interest has grown in therapeutic inhibition of NF-kappaB to limit inflammatory injury during sepsis. However, NF-kappaB also regulates protective responses, and predicting the net survival effects of such inhibition may be difficult. OBJECTIVES: To highlight the caution necessary with this therapeutic approach, we review our investigations in a mouse sepsis model with parthenolide and ethyl pyruvate, two NF-kappaB inhibitors proposed for clinical study. RESULTS: Consistent with published studies, parthenolide decreased NF-kappaB binding activity and inflammatory cytokine release from lipopolysaccharide (LPS) stimulated RAW 264.7 cells in vitro. In LPS-challenged mice (C57BL/6J), however, while both agents decreased lung and kidney NF-kappaB binding activity and plasma cytokines early (1-3 h), these measures were increased later (6-12 h) in patterns differing significantly over time. Furthermore, despite studying several doses of parthenolide (0.25-4.0 mg/kg) and ethyl pyruvate (0.1-100 mg/kg), each produced small but consistent decreases in survival which overall were significant (p < or = 0.04 for each agent). CONCLUSION: While NF-kappaB inhibitors hold promise for inflammatory conditions such as sepsis, caution is necessary. Clear understanding of the net effects of NF-kappaB inhibitors on outcome will be necessary before such agents are used clinically. PMID- 19555304 TI - New approach for local delivery of rapamycin by bioadhesive PLGA-carbopol nanoparticles. AB - Local delivery of antiproliferative drugs encapsulated in biodegradable nanoparticles has shown promise as an experimental strategy for preventing vascular restenosis development. The general aim of this work was to develop polymeric nanoparticle carriers with bioadhesive properties, and to evaluate its adjuvant potential for local, intramural delivery of rapamycin for inhibition of restenosis. The bioadhesive rapamycin-loaded PLGA nanoparticles were obtained by applying carbopol 940 of different concentrations as stabilizer and bioadhesive agent. The resultant nanoparticles were characterized concerning physicochemical properties such as morphology, particle size, zeta potential, entrapment efficiency, drug loading, drug release in vitro, stability in vitro as well as the arterial uptake and retention ability in an ex-vivo model. The results revealed that carbopol could serve as a better stabilizer in the preparation of rapamycin-loaded PLGA nanoparticles compared with PVA, and the physicochemical characteristics of the obtained PLGA nanoparticles were affected by the concentration of carbopol. Furthermore, it was found that carbopol could impart the nanoparticles with bioadhesive properties, improving the rentention and uptake of nanoparticles in the arterial wall, benefiting the nanoparticles for efficient localization of therapeutic agents in restenosis site. Cell viability assay results showed that blank PLGA-carbopol nanoparticles exhibited low toxicity and excellent biocompatibility and rapamycin-loaded nanoparticles with a smaller particle size (< 200 nm) had an increased antiproliferative effect on cells in comparison to free drug. These results indicated that this research might provide a potential experimental basis for the further study of carbopol stabilized bioadhesive nanoparticles against restenosis in vivo. PMID- 19555292 TI - Advances in light microscopy for neuroscience. AB - Since the work of Golgi and Cajal, light microscopy has remained a key tool for neuroscientists to observe cellular properties. Ongoing advances have enabled new experimental capabilities using light to inspect the nervous system across multiple spatial scales, including ultrastructural scales finer than the optical diffraction limit. Other progress permits functional imaging at faster speeds, at greater depths in brain tissue, and over larger tissue volumes than previously possible. Portable, miniaturized fluorescence microscopes now allow brain imaging in freely behaving mice. Complementary progress on animal preparations has enabled imaging in head-restrained behaving animals, as well as time-lapse microscopy studies in the brains of live subjects. Mouse genetic approaches permit mosaic and inducible fluorescence-labeling strategies, whereas intrinsic contrast mechanisms allow in vivo imaging of animals and humans without use of exogenous markers. This review surveys such advances and highlights emerging capabilities of particular interest to neuroscientists. PMID- 19555305 TI - Gastroresistant microcapsules: new approaches for site-specific delivery of ketoprofen. AB - Ketoprofen is a potent non-steroidal anti-inflammatory drug (NSAID) that has been widely used in the treatment of rheumatoid arthritis and other related conditions. However, it carries the risk of undesirable systemic side effects and gastrointestinal irritation at the usual dose of oral administration. The aim of this study was to prepare and evaluate gastroresistant microcapsules containing ketoprofen. Microcapsules were obtained by a spray-drying process starting from an O/A emulsion in the presence of different pH-dependent materials (Eudragit L100, Eudragit S100, and stearic acid) dissolved in the external phase. The influence of formulation factors (oily phase employed for drug solubilization, type of coating) on the morphology, particle size distribution, drug loading capacity, in-vitro release, and ex-vivo permeation characteristics were investigated. Drug loading capacity was very high for all the microcapsules prepared. Formulation factors did not significatively influence the mean particle size, but modified microcapsule in-vitro and ex-vivo behavior. PMID- 19555306 TI - Preparation, characterization and in vitro intestinal absorption of a dry emulsion formulation containing atorvastatin calcium. AB - A redispersible dry emulsion (DE) formulation of atorvastatin calcium (AC) was developed to enhance the in vitro dissolution of AC, thereby increasing its gastrointestinal absorption. The spray-drying technology was used where Plurol Oleique CC 497 was chosen as the oil phase. Effects of carriers, surfactants, and homogenizers on the characteristics of DE containing AC were systematically investigated. The final formulation consisted of dextrin and Poloxamer 188 as carrier and surfactant, respectively, and was homogenized by a high pressure homogenizer before spray drying. The in vitro release of AC from the optimized DE was significantly higher than that of pure AC powder (76% vs. 30% at 24 hr). The in vitro intestinal absorption of AC from the DE formulation was 0.77 microg/cm(2) at 2 hr, which was a 2.33-fold increase compared to the pure unformulated AC powder. These results suggest that the oral dry emulsion formulation could improve the intestinal absorption of AC. PMID- 19555307 TI - Preparation and performance evaluation of saquinavir laden cationic submicron emulsions. AB - The main purpose of the present study was to investigate different cationic submicron emulsions as potential delivery for oral administration. Different submicron emulsion based formulations were prepared by standard procedures incorporating Chitosan, stearylamine, and protamine as charge inducer. Saquinavir (SQ) laden emulsions were characterized in terms of globule size, zeta potential, entrapment efficiency, release profile, cytotoxicity, LDH release, and stability studies. The prepared formulations were stable in terms of mean globule size, drug content, and tended to retain their cationic charge. Pay load efficiency was found to be pretty high (approximately 95-99%) in various formulations prepared. Sustained release phenomenon was more prominent in the case of chitosan emulsions (CE) followed by stearylamine emulsion (SE), Protamine emulsion (PE), and then plain emulsion (E) containing no charge inducer. The total amounts of drug released in 24 hr from CE, SE, PE, and E were 46%, 52%, 56%, and 62%, respectively. The induction of positive charge in emulsions resulted in enhanced absorption of drug through intestinal membrane. The apparent permeability coefficient through the intestinal sac was in the order of CE > SE > PE > E. The permeation flux of SQ through CE (1.0 microg/min) was more than twice compared to plain emulsion (0.46 microg/min) while it was almost three times (0.3 microg/min) compared to control. However, protamine based emulsion didn't confer significant improvement in absorption when compared to plain emulsion formulation. By this study it can be concluded that induction of positive charge on submicron emulsions can be effective for improving oral absorption of drug safely, as it is evinced with low LDH release into the medium when intestinal tissue is treated with submicron emulsion. PMID- 19555310 TI - Controlled-release carbamazepine matrix granules and tablets comprising lipophilic and hydrophilic components. AB - The objective of this study was to investigate the effect of lipophilic (Compritol 888 ATO) and hydrophilic components (combination of HPMC and Avicel) on the release of carbamazepine from granules and corresponding tablet. Wet granulation followed by compression was employed for preparation of granules and tablets. The matrix swelling behavior was investigated. The dissolution profiles of each formulation were compared to those of Tegretol CR tablets and the mean dissolution time (MDT), dissolution efficiency (DE%), and similarity factor (f(2) factor) were calculated. It was found that increase in the concentration of HPMC results in reduction in the release rate from granules and achievement of zero order is difficult from the granules. The amount of HPMC plays a dominant role for the drug release. The release mechanism of CBZ from matrix tablet formulations follows non-Fickian diffusion shifting to Case II by the increase of HPMC content, indicating significant contribution of erosion. Increasing in drug loading resulted in acceleration of the drug release and in anomalous controlled release mechanism due to delayed hydration of the tablets. These results suggest that wet granulation followed by compression could be a suitable method to formulate sustained release CBZ tablets. PMID- 19555309 TI - Effect of microneedle on the pharmacokinetics of ketoprofen from its transdermal formulations. AB - Non-invasive transdermal delivery using microneedle arrays was recently introduced to deliver a variety of large and hydrophilic compounds into the skin, including proteins and DNA. In this study, a microneedle array was applied to the delivery of a hydrophobic drug, ketoprofen, to determine if transdermal delivery in rats can be improved without the need for permeation enhancers. The ability of a microneedle to increase the skin permeability of ketoprofen was tested using the following procedure. A microneedle array was inserted into the lower back skin of a rat using a clip for 10 min. Subsequently, 24 mg/kg of a ketoprofen gel was loaded on the same site where the microneedle had been applied. Simultaneously, the microneedle was coated with 24 mg/kg of a ketoprofen gel, and inserted into the skin using a clip for 10 min. As a negative control experiment, only 24 mg/kg of the ketoprofen gel was applied to the shaved lower back of a rat. Blood samples were taken at the indicated times. The plasma concentration (C(p)) was obtained as a function of time (t), and the pharmacokinetic parameters were calculated using the BE program. The group loaded with the microneedle coated with ketoprofen gel showed a 1.86-fold and 2.86-fold increase in the AUC and C((max)) compared with the ketoprofen gel alone group. These results suggest that a microneedle can be an ideal tool for transdermal delivery products. PMID- 19555311 TI - Epidemiology and correlates of weight worry in the multinational Diabetes Attitudes, Wishes and Needs study. AB - OBJECTIVE: This study examined the epidemiology of worry about weight among adults with diabetes and whether this weight worry was associated with worse quality of life and diabetes self-management. RESEARCH DESIGN AND METHODS: Participants were 5088 adults with type 1 or type 2 diabetes from the multi national DAWN study. Random samples of approximately 500 were obtained from each of 11 countries/regions in Asia, Australia, Europe, and North America. All measures were self-reported data obtained during structured interviews. High weight worry was represented by the highest level of agreement with the statement 'I feel very anxious about my weight.' RESULTS: One-quarter (25.1%) of respondents reported high weight worry. Using multivariate logistic regression to assess independent relationships, high weight worry was significantly (p < 0.05) more common among respondents who were women, less educated, had type 2 diabetes and more comorbidities, were diagnosed more recently and reported weight gain with diabetes. Using multivariate logistic regression to control for confounding factors, high weight worry was significantly (p < 0.05) associated with most of the adverse outcomes examined, including lower self-rated health, poorer reported regimen adherence and diabetes control, more diabetes-related distress, poorer psychological well-being, and more psychological treatment. LIMITATIONS: The cross-sectional, correlational analysis could not assess causal relationships among patient reported outcomes, and the study did not include objective outcome measures. CONCLUSIONS: High weight worry was common among adults with diabetes and associated with several adverse outcomes, including multiple indicators of diabetes-related distress, poor physical and psychological well-being, and regimen non-adherence. These findings suggest that healthcare providers should assess and address weight worry among their patients with diabetes. PMID- 19555312 TI - Role and significance of statins in the treatment of hypertensive patients. AB - BACKGROUND: Statins are the first-line drug therapy in the treatment of hypercholesterolemia. The beneficial clinical impact of statins on the cardiovascular system results not only from their lipid-lowering action but also from other effects. Recently, it has been suggested that statins can reduce blood pressure, especially in hypertensive patients. AIM: The role of the hypotensive action of statins and other mechanisms which reduce cardiovascular risk in hypertensive patients are discussed in this review. METHODS: Electronic databases searched were [MEDLINE (1966 - February 2009), EMBASE and SCOPUS (1965 - February 2009), DARE (1966 -- February 2009)]. Additionally, abstracts from national and international cardiovascular meetings were studied to identify unpublished studies. The main data search terms were: blood pressure, hypertension, hypercholesterolemia and statins. FINDINGS: At present, it is difficult to unequivocally assess the impact of statins on blood pressure. However, according to most authors, the impact of statins on the decrease in BP is slight, but significant, especially among patients with hypertension. PMID- 19555308 TI - In vitro cytotoxicity of novel pro-apoptotic agent DM-PIT-1 in PEG-PE-based micelles alone and in combination with TRAIL. AB - The purpose of this study was to develope and characterize a micellar formulations of N-{[(2-hydroxy-5- nitrophenyl)amino]carbonothioyl}-3,5 dimethylbenzamide (DM-PIT-1)-a new small molecule non-lipid antagonist of phopshotidylinositol-3.4.5-triphopshate and inhibitor of the PI3-kinase pathway. Micelle-forming PEG(2000)-PE was used to solubilize DM-PIT-1. To improve the specificity of the micellar DM-PIT-1, cancer-targeting anti-nucleosomal mAb2C5 antibodies as well as Tumor necrosis factor- Related Apoptosis-Inducing Ligand (TRAIL) were attached to the surface of polymeric micelles. DM-PIT-1 was effectively incorporated (> 70%) into 14-16 nm micelles, which had a negative surface zeta potential of 4-5 mV. Micellar DM-PIT-1 demonstrated high in vitro cytotoxicity against various cancer cells. An improved potency of the dual activity DM-PIT-1/TRAIL combination nanoparticles in inducing death of TRAIL resistant cancer cells was shown. Efficacy of the TRAIL therapy was enhanced by combining it with the 2C5 antibody cancer-targeted micellar form of DM-PIT-1. In conclusion, DM-PIT-1 micellar preparations can be used for targeted combination therapy against TRAIL-resistant cancers. PMID- 19555314 TI - Physiological variables involved in heart valve substitute calcification. AB - Biochemical, histological and genetic studies using in vitro/in vivo models have demonstrated that pathological calcification of bioprosthetic heart valves (BHV) is regulated by various mechanisms associated with physiological variables. The major objective of this review is to characterize physiological variables involved in BHV calcification. This review examines our understanding of the systemic cellular behavior and physiological regulation processes behind BHV calcification and its clinical applications. PMID- 19555313 TI - Prepandemic H5N1 influenza vaccine adjuvanted with AS03: a review of the pre clinical and clinical data. AB - BACKGROUND: Universal and timely administration of a prepandemic vaccine is considered to be one of the most effective measures to reduce the incidence of pandemic influenza infection and consequently its morbidity and mortality. OBJECTIVES: To provide the reader with basic insights into influenza virus infections, the threat of a pandemic and the challenges it poses for vaccine development. METHODS: This review summarizes the reported preclinical and clinical data obtained with the prepandemic H5N1 vaccine adjuvanted with AS03. RESULTS: The AS03-adjuvanted prepandemic H5N1 influenza vaccine allows for antigen sparing, has a good safety and acceptable reactogenicity profile, induces an immune response that not only meets all European Committee for Medicinal Products (CHMP) and FDA requirements for the vaccine strain but also generates neutralizing antibodies that broadly cross-react against H5N1 drift strains, and finally conveys protection in a ferret model against lethal challenges with homologous and heterologous H5N1 viruses. PMID- 19555315 TI - Effect of continuous silymarin administration on oral talinolol pharmacokinetics in healthy volunteers. AB - 1. The objective of this study was to investigate the effect of concomitantly administered silymarin on the pharmacokinetics of talinolol, a typical substrate for P-glycoprotein (P-gp), in healthy Chinese volunteers and its association with a multidrug resistance 1 (MDR1) C3435T genetic polymorphism. 2. Eighteen healthy adult men (six MDR1 3435CC homozygotes, six MDR1 3435CT heterozygotes and six MDR1 3435TT homozygotes) were recruited in a two-phase, randomized, single-blind, crossover design. The pharmacokinetics of talinolol were measured after co administration of placebo or 140 mg silymarin capsules three times daily for 14 days. Concentrations of talinolol in plasma were measured for up to 36 h after drug administration by liquid chromatography-mass spectrometry (HPLC-MS). 3. The peak plasma concentration (C(max)) of talinolol was significantly higher after silymarin administration as compared with placebo (p = 0.007). The area under the plasma concentration-time curve from zero to 36 h (AUC(0-36)) and AUC(0-infinity) of talinolol was increased by 36.2% +/- 33.2% and 36.5% +/- 37.9%, respectively, by silymarin co-administration. The oral clearance (CL/F) of talinolol was decreased by 23.1% +/- 16.6% (p < 0.001) during the silymarin-treated phase. No change in the time to peak concentration (t(max)) and the blood elimination half life (t(1/2)) of talinolol was observed between the placebo- and silymarin treated phases. 4. Co-administration of silymarin significantly increased the plasma concentration of talinolol in healthy volunteers. PMID- 19555348 TI - Methylation status of oestrogen receptor alpha-A: a predictor of prognosis in leukaemias. AB - Many studies have shown that epigenetic regulation of ERs (oestrogen receptors) plays a key role in the pathogenesis of leukaemia. In the present study, it was found that the methylated status of ERalpha-A might serve as an epigenetic biomarker of leukaemias. In this study, the protein expression and cell apoptosis, cycle, proliferation and viability with and without 5-aza-dC (5-aza-2' deoxycytidine) were evaluated with Western blotting, 3H-TdR (3H-thymidine) incorporation, propidium iodide staining and Trypan Blue staining respectively. The protein expression of ERalpha was significantly enhanced in all leukaemic cell lines using treatment with the DNA demethylation reagent 5-aza-dC. However, no obvious change in the protein expression of ERbeta takes place with 5-aza-dC. And with 5-aza-dC, cell apoptosis, cell cycle, cell proliferation and viability were all inhibited significantly. We also tracked 40 cases of leukaemias with ERalpha-A methylation (95%; 38 of 40) to observe the prognosis 1 year after chemotherapy treatment. The patients with ERalpha-A methylation have no obvious symptomatic relief; however, two patients without ERalpha-A methylation have obtained effective relief. This result suggested that ERalpha plays a significant role in leukaemogenesis, and the methylated status of ERalpha-A not only might serve as an epigenetic biomarker of leukaemias for diagnosis, but also has the potential to serve as a predictor of prognosis in leukaemias. PMID- 19555349 TI - The transcriptional activity of Pygopus is enhanced by its interaction with cAMP response-element-binding protein (CREB)-binding protein. AB - Pygopus is a core component of the beta-catenin/TCF (T-cell factor) transcriptional activation complex required for the expression of canonical Wnt target genes. Recent evidence suggests that Pygopus could interpret histone methylation associated with target genes and it was shown to be required for histone acetylation. The involvement of a specific acetyltransferase, however, was not determined. In this report, we demonstrate that Pygopus can interact with the HAT (histone acetyltransferase) CBP [CREB (cAMP-responsive-element-binding protein)-binding protein]. The interaction is via the NHD (N-terminal homology domain) of Pygopus, which binds to two regions in the vicinity of the HAT domain of CBP. Transfected and endogenous hPygo2 (human Pygopus2) and CBP proteins co immunoprecipitate in HEK-293 (human embryonic kidney 293) cells and both proteins co-localize in SW480 colorectal cancer cells. The interaction with CBP also enhances both DNA-tethered and TCF/LEF1 (lymphoid enhancing factor 1)-dependent transcriptional activity of Pygopus. Furthermore, immunoprecipitated Pygopus protein complexes displayed CBP-dependent histone acetyltransferase activity. Our data support a model in which the NHD region of Pygopus is required to augment TCF/beta-catenin-mediated transcriptional activation by a mechanism that includes both transcriptional activation and histone acetylation resulting from the recruitment of the CBP histone acetyltransferase. PMID- 19555350 TI - Functional conservation of tRNase ZL among Saccharomyces cerevisiae, Schizosaccharomyces pombe and humans. AB - Although tRNase Z from various organisms was shown to process nuclear tRNA 3' ends in vitro, only a very limited number of studies have reported its in vivo biological functions. tRNase Z is present in a short form, tRNase Z(S), and a long form, tRNase Z(L). Unlike Saccharomyces cerevisiae, which contains one tRNase Z(L) gene (scTRZ1) and humans, which contain one tRNase Z(L) encoded by the prostate-cancer susceptibility gene ELAC2 and one tRNase Z(S), Schizosaccharomyces pombe contains two tRNase Z(L) genes, designated sptrz1(+) and sptrz2(+). We report that both sptrz1(+) and sptrz2(+) are essential for growth. Moreover, sptrz1(+) is required for cell viability in the absence of Sla1p, which is thought to be required for endonuclease-mediated maturation of pre-tRNA 3' ends in yeast. Both scTRZ1 and ELAC2 can complement a temperature sensitive allele of sptrz1(+), sptrz1-1, but not the sptrz1 null mutant, indicating that despite exhibiting species specificity, tRNase Z(L)s are functionally conserved among S. cerevisiae, S. pombe and humans. Overexpression of sptrz1(+), scTRZ1 and ELAC2 can increase suppression of the UGA nonsense mutation ade6-704 through facilitating 3' end processing of the defective suppressor tRNA that mediates suppression. Our findings reveal that 3' end processing is a limiting step for defective tRNA maturation and demonstrate that overexpression of sptrz1(+), scTRZ1 and ELAC2 can promote defective tRNA 3' processing in vivo. Our results also support the notion that yeast tRNase Z(L) is absolutely required for 3' end processing of at least a few pre-tRNAs even in the absence of Sla1p. PMID- 19555352 TI - Apoptolysis: a novel mechanism of skin blistering in pemphigus vulgaris linking the apoptotic pathways to basal cell shrinkage and suprabasal acantholysis. AB - Understanding the acantholytic pathways leading to blistering in pemphigus vulgaris (PV) is a key to development of novel treatments. A novel paradigm of keratinocyte damage in PV, termed apoptolysis, links the suprabasal acantholytic and cell death pathways to basal cell shrinkage rendering a 'tombstone' appearance to PV lesions. In contrast to apoptolysis, the classic keratinocyte apoptosis mediating toxic epidermal necrolysis causes death and subsequent sloughing of the entire epidermis. Apoptolysis includes five consecutive steps. (1) Binding of autoantibodies to PV antigens. (2) Activation of EGF receptor, Src, mTOR, p38 MAPK and other signalling elements downstream of ligated antigens, elevation of intracellular calcium and launching of the cell death cascades. (3) Basal cell shrinkage due to: (i) collapse and retraction of the tonofilaments cleaved by executioner caspases; and (ii) dissociation of interdesmosomal adhesion complexes caused by phosphorylation of adhesion molecules. (4) Massive cleavage of cellular proteins by activated cell death enzymes leading to cell collapse, and tearing off desmosomes from the cell membrane stimulating secondary autoantibody production. (5) Rounding up and death of acantholytic cells. Thus, the structural damage (acantholysis) and death (apoptosis) of keratinocytes are mediated by the same cell death enzymes. Appreciation of the unifying concept of apoptolysis have several important implications: (i) linking together a number of seemingly unrelated events surrounding acantholysis; (ii) opening new avenues of investigation into the pathomechanism of pemphigus; and (iii) creating new approaches to the treatment of pemphigus based on blocking the signalling pathways and enzymatic processes that lead to blistering. PMID- 19555353 TI - Identification of the etiologies of chronic urticaria in children: a prospective study of 94 patients. AB - The etiologies of chronic urticaria (CU) in childhood remains incompletely understood because of limited data in children. The objective of this study was to examine some of the possible etiologies of CU in children by focusing on the functional autoantibody to FcepsilonRIalpha and IgE, thyroid autoimmunity, urticarial vasculitis, parasitic infestation and food allergy. Children 4-15 yr of age with CU were investigated for complete blood count, erythrocyte sedimentation rate (ESR), antinuclear antibody (ANA), CH(50), free-T4 (FT(4)), thyroid stimulating hormone (TSH), anti-thyroglobulin and anti-microsomal antibody, autologous serum skin test (ASST), skin prick tests (SPT) for foods, food challenges, and stool examination for parasites. Ninety-four children who met the criteria for CU were recruited. Patients with physical urticaria were excluded. Eosinophilia and elevated ESR were found in 23% and 13%, respectively. High ANA titers were found in 2%. None of these patients had clinical features of urticarial vasculitis, abnormal CH(50) level, abnormal TSH and FT(4). Anti thyroglobulin and anti-microsomal antibodies were not detected. Positive ASST was found in 38%. There were no differences in medication requirement and CU remission between patients with positive and negative ASST. Parasites were found in 5% without clinical correlation. SPT to foods was positive in 35%. Positive food challenges were found in six/nine patients with positive history of food allergy and two/seven patients with negative history. Food avoidance was beneficial to the subgroup of patients with positive history of food allergy only. PMID- 19555354 TI - Cow's milk allergy related pediatric constipation: appropriate time of milk tolerance. AB - Cow's milk allergy (CMA) has different clinical presentations according to age. The study aimed to evaluate the extent of CMA as a cause of pediatric constipation and determine the appropriate timing of tolerance to cow milk in such patients. The study included 60 patients suffering from chronic functional constipation, 27 of whom did not respond to 2 month laxative therapy (group I). Thirty age and sex matched apparently healthy infants and children were studied as a control group (group II). Serum specific IgE to cow milk proteins was measured. Withdrawal of cow milk and dairy products for a 1 month period was then followed by cow milk re-challenge over 2 wk. Patients were classified into: responders to this schedule (cow milk allergic=group Ia; n=21) and non-responders (non-cow milk allergic=group Ib; n=6). Eighteen CMA patients continued the study where nine of them underwent milk reinstitution after 6 months and another nine patients after 12 months of elimination. The frequency of CMA among constipated patients was 77.7%. Mean values of serum specific IgE to whole cow milk protein and beta-lactoglobulins were significantly higher in constipated patients (0.82+/ 0.08, 0.79+/-0.13 IU/ml, respectively) compared with controls (0.26+/-0.14, 0.27+/-0.14 IU/ml, respectively) and in group Ia (0.99+/-0.08, 0.95+/-0.14 IU/ml, respectively) compared with group Ib (0.39+/-0.06, 0.37+/-0.10 IU/ml, respectively). Serum specific IgE was positive in 85.7% of CMA group, predominantly in class 2. Tolerance to cow milk was achieved after 6 months in only 22.2% compared with 88.8% after 12 months of elimination. In conclusion, CMA is shown to be a significant etiologic factor for constipation in infants and young children. Serum levels of IgE to cow milk proteins are helpful although not definitive for diagnosis. Based on this limited study, tolerance is better achieved after 12 months of strict cow's milk elimination. PMID- 19555351 TI - Sodium nitrite therapy attenuates the hypertensive effects of HBOC-201 via nitrite reduction. AB - Hypertension secondary to scavenging of NO remains a limitation in the use of HBOCs (haemoglobin-based oxygen carriers). Recent studies suggest that nitrite reduction to NO by deoxyhaemoglobin supports NO signalling. In the present study we tested whether nitrite would attenuate HBOC-mediated hypertension using HBOC 201 (Biopure), a bovine cross-linked, low-oxygen-affinity haemoglobin. In a similar way to unmodified haemoglobin, deoxygenated HBOC-201 reduced nitrite to NO with rates directly proportional to the extent of deoxygenation. The functional importance of HBOC-201-dependent nitrite reduction was demonstrated using isolated aortic rings and a murine model of trauma, haemorrhage and resuscitation. In the former, HBOC-201 inhibited NO-donor and nitrite-dependent vasodilation when oxygenated. However, deoxygenated HBOC-201 failed to affect nitrite-dependent vasodilation but still inhibited NO-donor dependent vasodilation, consistent with a model in which nitrite-reduction by deoxyHBOC-201 counters NO scavenging. Finally, resuscitation using HBOC-201, after trauma and haemorrhage, resulted in mild hypertension ( approximately 5-10 mmHg). Administration of a single bolus nitrite (30-100 nmol) at the onset of HBOC-201 resuscitation prevented hypertension. Nitrite had no effect on mean arterial pressure during resuscitation with LR (lactated Ringer's solution), suggesting a role for nitrite-HBOC reactions in attenuating HBOC-mediated hypertension. Taken together these data support the concept that nitrite can be used as an adjunct therapy to prevent HBOC-dependent hypertension. PMID- 19555355 TI - Glass transition temperature of hard chairside reline materials after post polymerisation treatments. AB - OBJECTIVE: This study evaluated the effect of post-polymerisation treatments on the glass transition temperature (T(g)) of five hard chairside reline materials (Duraliner II-D, Kooliner-K, New Truliner-N, Ufi Gel hard-U and Tokuso Rebase Fast-T). MATERIALS AND METHODS: Specimens (10 x 10 x 1 mm) were made following the manufacturers' instructions and divided into three groups (n = 5). Control group specimens were left untreated. Specimens from the microwave group were irradiated with pre-determined power/time combinations, and specimens from the water-bath group were immersed in hot water at 55 degrees C for 10 min. Glass transition ( degrees C) was performed by differential scanning calorimetry. Data were analysed using anova, followed by post hoc Tukey's test (alpha = 0.05). RESULTS: Both post-polymerisation treatments promoted a significant (p < 0.05) increase in the T(g) of reline material K. Materials K, D and N showed the lowest T(g) (p < 0.05). No significant difference between T and U specimens was observed. CONCLUSION: Post-polymerisation treatments improved the glass transition of material Kooliner, with the effect being more pronounced for microwave irradiation. PMID- 19555356 TI - Understanding the 'epidemic' of complete tooth loss among older New Zealanders. AB - OBJECTIVE: The aim of this study was to obtain a deeper understanding of the social factors driving New Zealand's historic 'epidemic of edentulism' and how they operated. METHOD: In-depth, semi-structured interviews with 31 older New Zealanders were analysed using applied grounded theory. RESULTS: Universal factors present in the data were: (a) the way in which New Zealand society accepted and indeed encouraged edentulism without stigma for those who had a 'sub optimal' natural dentition; (b) how the predominant patterns of dental care utilisation (symptomatic and extraction-based) were often strongly influenced by economic and social disadvantage; and (c) the way in which lay and professional worldviews relating to 'calcium theory' and dental caries were fundamental in decisions relating to the transition to edentulism. Major influences were rural isolation, the importance of professional authority and how patient-initiated transitions to edentulism were ultimately facilitated by an accommodating profession. CONCLUSION: The combined effects of geography, economics, the dental care system and the professional culture of the day, in the context of contemporary (flawed) understandings of oral disease, appear to have been the key drivers. These were supported (in turn) by a widespread acceptance by the profession and society alike of the extraction/denture philosophy in dealing with oral disease. PMID- 19555357 TI - Interleukin-6 (G-174C) and tumour necrosis factor-alpha (G-308A) gene polymorphisms in geriatric patients with chronic periodontitis. AB - BACKGROUND AND OBJECTIVE: Periodontitis is a chronic inflammatory disease, and genetic factors may have an important role in its severity. Polymorphisms in the promoter regions of the interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha) genes have been reported to cause changes in the production of these cytokines. The aim of this study was to evaluate the possible role of IL-6 (G 174C) and tumour necrosis factor (G-308A) polymorphisms, in the severity of chronic periodontitis in an elderly population. MATERIALS AND METHODS: In this study, a group of 65 elderly women, comprising 17 patients with moderate chronic periodontitis, 21 with severe chronic periodontitis and 27 healthy patients were selected. DNA was isolated from all subjects, and polymerase chain reaction was used to study the IL-6 and TNF-alpha gene polymorphisms. RESULTS: The results of this study showed a significant difference in the allele and genotype frequencies of IL-6 gene polymorphism between patients with periodontal disease and controls. Subjects carrying the G/G genotype of IL-6 were most severely affected by periodontitis. The TNF-alpha gene polymorphism showed no association with chronic periodontitis between patients and controls. CONCLUSION: The results suggest that the IL-6 gene polymorphism may be associated with chronic periodontitis, and that TNF-alpha gene polymorphism may not be involved in the progression of chronic periodontitis in the population of elderly Brazilian women. PMID- 19555358 TI - Assessment of mobile dental services in the State of Lower Austria, Austria. AB - OBJECTIVES: To investigate the existence and the quality of mobile dental services available to the austrian elderly population of Lower Austria, Austria, using two approaches: the home-bound elders' perception and the dentistry services provided in nursery homes. METHODS: The study had two parts. The first was a qualitative assessment of mobile dental services as perceived by the elderly home bounds, based on 52 questionnaires. These questionnaries were distributed to the home bound persons in a period of one month. The second part was the assessment in the nursing homes. For this study a short questionnaire was sent, by e-mail, to 61 nursing homes of Lower Austria. RESULTS: by the elderly home bound--29 questionnaires returned completed (response quote of 55.8%) and 4 uncompleted. The age range of the subjects was from 64 to 89 years old and the ratio of women to men was about 2:1 (62% to 38%). 79.3% think that dental home care is a good practice, 58.6% would use mobile dentistry themselves. in the nursing homes--From the 61 sent e-mails 33 were returned with completed questionnaires (response quote of 54.09%). 48.5% of the nursing homes replied that they do not have mobile dental service. However, 17 homes (51.51%) have a mobile dentist. CONCLUSION: A mobile dental service for patients of austrian geriatric nursing homes does exist. However, no mobile dental service was identified for austrian home bound patients. This scenario shows the need for a mobile dental care programme and politcs. PMID- 19555359 TI - Oral health and oral implant status in edentulous patients with implant-supported dental prostheses who are receiving long-term nursing care. AB - AIM: The aim of this study was to investigate oral health and oral implant status in a group of edentulous patients receiving long-term residential or nursing care (LTC), all of whom had implant-supported fixed or removable dental prostheses. MATERIAL AND METHODS: A dental examination was performed on a total of 3310 patients receiving LTC and from this population 35 edentulous patients in whom dental implants had been placed formed the cohort for this study. All examinations were performed by a specialist in hospital dentistry and took place in the patients' own home environment. Oral health was assessed by means of a protocol which evaluated oral hygiene status, possible oral mucosal inflammation and oral mucosal friction levels. Any problems with the implant-supported prosthesis, implant mobility or other complications were also assessed. In addition, patients were asked about any oral symptoms and their usual oral hygiene procedures. RESULTS: About half of the subjects (17/35) were registered as having no/mild inflammation with 18 of 35 having moderate/severe inflammation. Twelve of the 35 patients had good/acceptable oral hygiene and 23 of 35 had poor/bad oral hygiene. Twenty-one of the 35 patients depended on help from the nursing personnel for their daily oral hygiene procedures. Obvious problems with food impaction were noted in 11 patients. A total of 229 implants had been placed in 43 jaws supporting 40 full arch-fixed prostheses and three implant-borne overdentures. There was no evidence of mobility or fractures of either the implants or the prostheses. Fifteen implants showed some exposed screw threads. Pus was exuding from one implant site and general peri-implant gingival hyperplasia was noted in two patients. Twenty-four patients were completely satisfied with the function and appearance of their implant-supported prostheses. Two patients were totally dissatisfied. CONCLUSION: This study indicates that oral implant therapy can be considered as a treatment of choice in elderly patients, even if oral hygiene is sub-optimal. PMID- 19555360 TI - Evaluation of an oral function promotion programme for the independent elderly in Japan. AB - OBJECTIVES: The purpose of this study was to provide an oral function promotion programme for the independent elderly and evaluate the changes in oral health status and oral function. BACKGROUND: Few studies have scientifically analysed and evaluated the effectiveness of oral function promotion programmes provided for the independent elderly. MATERIALS AND METHODS: The subjects were independent elderly females (mean age: 74.6 +/- 6.3) recruited from senior citizens' centres in Tokyo. The intervention group (n = 79) received a 3-month oral function promotion programme, which included facial muscle and tongue exercises and salivary gland massages. The control group (n = 62) did not receive this programme. RESULTS: In the intervention group, the tongue coating scores decreased and the organoleptic score of oral malodour fell. The amount of food debris in the oral cavity decreased and the tongue dryness improved. Furthermore, the salivary flow rate increased. The length of time for maintaining the tongue in the forward position increased from 11.2 s to 18.7 s, and the number of times for moving the tip of the tongue in a clockwise circular motion, counter clockwise circular motion and side-to-side motion within 30 s, increased from 14.5 to 20.6, 14.5 to 20.2, and 17.2 to 23.3 respectively. The number of times for movement of the lips significantly improved from 23.0 to 28.8 and the pronunciation of words was observed to be clearer. CONCLUSION: An oral function promotion programme was effective in improving the oral health status and oral function of an independent elderly population. PMID- 19555362 TI - Factors associated with remaining teeth of elderly in the state of Sao Paulo, Brazil, 2002. AB - Objectives: The objective of this study was to verify the association between the number of teeth present with socio-demographic and economic variables and with the access to dental services and self-perception of oral health among the elderly. Materials and Methods: The sample was composed of individuals from 65 to 74 years of age, which was representative of the state of Sao Paulo (n = 781). In this study, the analysis of data considered dentate elderly only, who were divided into two groups: those with one to 19 teeth and those with 20 teeth or more. The chi-squared test was used in the bivariate analysis and the logistic regression was also performed (p < 0.05). Results: Among the 313 dentate elderly, 235 (75.1%) presented one to 19 teeth. The average number of teeth was 9.5 for the elderly with fewer teeth and 25 for those with more teeth. In the bivariate analysis, all variables related to the access to dental services were associated and the chances of having less teeth was among elderly who did not visit the dentist for a long time and among those who sought public dental services and did not receive information on how to avoid oral problems. In addition, the elderly who classified their speech as not good were those who presented with fewer teeth. Conclusion: Most subjects presented one to 19 teeth and this fact was directly associated, among others factors, to the unsatisfactory perception that they reported in relation to their speech and to the too long period without visiting the dentist. PMID- 19555361 TI - Home or away? Differences between home- and clinic-based dental examinations for older people. AB - OBJECTIVE: To determine whether home-based and clinic-based dental examinations of older people are comparable. BACKGROUND: Despite a number of studies which have examined the concordance between different examiners or examination circumstances, none has directly compared an oral examination conducted at home with one conducted in an appropriate clinical setting. MATERIALS AND METHODS: Dentate participants (n = 61) aged between 65 and 74 years underwent two clinical examinations, one in a conventional dental clinic and the other in the person's home. Kappa statistics, intra-class correlation coefficients and 'Difference against mean' plots were used to determine the nature and extent of any bias. RESULTS: The summary estimates for missing teeth and dental caries were close, although the home-based examinations resulted in a lower estimate of the mean number of decayed teeth and of untreated coronal decay. The lowest reliability statistics were observed with respect to the prevalence of untreated coronal caries, filled root surfaces and root surface (Decayed or Filled Surfaces) DFS. The periodontitis prevalence estimates were closer, but the reliability statistics were relatively low. The extent of bleeding on probing was relatively under-estimated in the home-based examinations. 'Difference against mean' plots indicated that, overall, the clinic-based examinations detected more disease (although this was not observed for all clinical parameters which were measured). CONCLUSION: While clinic-based examinations will remain the preferred option, the potential loss of information associated with home-based examinations is unlikely to be great enough to preclude using them where required, although surveys with larger samples (and therefore more accurate estimates) should restrict their proportion of home-based examinations to no more than 10%. As they are less precise, surveys with samples of 300 or fewer can safely accommodate up to about one-third of their examinations being conducted in participants' homes. PMID- 19555363 TI - Myelodysplastic syndromes: the role of flow cytometry in diagnosis and prognosis. AB - Flow cytometric immunophenotypic analysis is a well-recognized and widely accepted adjuvant test to increase the sensitivity and specificity of diagnosis in potential myelodysplastic syndrome (MDS) cases. In addition, flow cytometric analysis provides prognostic information in MDS that is not available from other sources. The flow cytometric findings indicative of MDS and its value is a diagnostic adjunct are discussed. PMID- 19555364 TI - Immunocryosurgery for basal cell carcinoma: results of a pilot, prospective, open label study of cryosurgery during continued imiquimod application. AB - BACKGROUND/AIM: Theoretical considerations support the combination of cryosurgery and topical imiquimod to treat basal cell carcinomas (BCC). The aim of the present study was to test the feasibility and efficacy of 'cryosurgery during continued imiquimod application' ('immunocryosurgery') to treat 'high-risk-for recurrence' BCCs. METHODS: Thirteen patients with 21 biopsy-proven tumours (4 of 21 relapses after prior surgery) were included. After 2-5 weeks (median, 3) of daily 5% imiquimod cream application, the tumours were treated by liquid N(2) cryosurgery (spray, two cycles, 10-20 s) and imiquimod was continued for additional 2-12 weeks (median, 4). The outcome after at least 18 months of follow up (18-24 months) is currently reported. RESULTS: Nineteen of 21 tumours responded promptly to immunocryosurgery; two tumours required additional treatment cycles to clear. Thus, the clinical clearance rate was 100%. Only 1 of 21(5%) tumour relapsed after at least 18 months of follow-up (cumulative efficacy: 95%). CONCLUSIONS: 'Immunocryosurgery' is a promising non-surgical combination modality to treat 'high-risk-for-recurrence BCCs'. Initial evidence is suggestive of an at least additive effect of the two combined modalities. Further studies comparing immunocryosurgery directly with cryosurgery and imiquimod monotherapies will confirm the reported results. PMID- 19555365 TI - Endonasal transsphenoidal surgery and multimodality treatment for giant pituitary adenomas. AB - OBJECTIVE: Giant pituitary adenomas (> or =40 mm) pose a major management challenge. We describe the experience of a single surgeon and a dedicated neuro endocrine team with multimodality treatment of these tumours in three specialized institutions. DESIGN: Retrospective data set analyses. PATIENTS: Fifty-one consecutive patients with a giant adenoma (39 endocrine-inactive, 12 endocrine active; mean tumour diameter 45 mm) treated over 10 years by an endonasal transsphenoidal approach were included. All patients had surgical resection followed by radiotherapy and/or medical therapy as judged necessary. MEASUREMENTS: Hormonal and visual status, extent of resection, tumour control rates, complications and use of medical and radiotherapy were evaluated. RESULTS: Surgery resulted in gross total, near total and subtotal removal in21 (41%), 10 (20%) and 20 (39%) patients respectively. Complete tumour removal was associated with absence of cavernous sinus invasion (P < 0.001). Long-term endocrine function improved in 49% of patients and new endocrinopathy occurred in 14.6%; 76% required long-term hormone replacement therapy. Vision improved in 81.5% of the patients and there was no visual worsening. At the last follow up (median 30 months), tumour control was achieved in 96% of patients: 59% with surgery alone, 20% with surgery plus focussed radiotherapy, 18% with surgery and medical therapy and two with all three modalities. CONCLUSIONS: Endonasal surgery provides effective initial treatment for patients with giant adenomas. Multimodality therapy was needed in almost 50% of patients and this rate will likely increase with longer follow up. Close collaboration of neurosurgeons with endocrinologists and radiation oncologists is essential for optimal treatment of patients with these challenging tumours. PMID- 19555366 TI - Indigenous hepatitis E virus infection of a plasma donor in Germany. AB - BACKGROUND: Although Europe is supposed to be non-endemic for hepatitis E virus (HEV), locally acquired human cases are registered, and a relatively high prevalence for anti-HEV was found in blood donors in some European countries. Transfusion-transmitted infections by contaminated blood products were reported in Japan and sporadically in Europe. MATERIALS AND METHODS: Several samples from a plasma donor were screened with a highly sensitive quantitative HEV real-time polymerase chain reaction and the full-length genome was generated. Serology was performed with two different commercially available ELISA kits. RESULTS: The full length genome sequence of human HEV was identified using samples from a plasma donor with acute self-limiting hepatitis. Plasma donated 2 weeks before onset of elevated liver enzyme levels was already positive for HEV RNA (10(4) copies/ml). High viraemia (10(6) copies/ml) correlated with the detection of anti-HEV IgM in the first blood sample with increased alanine transaminase levels. Phylogenetic analyses grouped the isolate within genotype 3, subtype 3f. CONCLUSION: The sequence analyses and the epidemiological data revealed that the plasma donor was most probably infected with a swine HEV. This case supports the ongoing discussion of an obligatory HEV nucleic acid testing of blood products for special recipient risk groups. PMID- 19555367 TI - Carbon balance of the oldest and most-shaded leaves in a vegetation: a litmus test for canopy models. PMID- 19555368 TI - The complete Glomus intraradices mitochondrial genome sequence--a milestone in mycorrhizal research. PMID- 19555369 TI - Presymbiotic factors released by the arbuscular mycorrhizal fungus Gigaspora margarita induce starch accumulation in Lotus japonicus roots. AB - * Nutrient exchange is the key symbiotic feature of arbuscular mycorrhiza (AM). As evidence is accumulating that plants sense presymbiotic factors from AM fungi and prepare for colonization, we investigated whether modifications in plant sugar metabolism might be part of the precolonization program. * Inoculation of Lotus japonicus roots in a double Millipore sandwich with the AM fungus Gigaspora margarita prevented contact between the symbionts but allowed exchange of signal molecules. Starch content was used as a marker for root carbohydrate status. * Mycorrhizal colonization of L. japonicus roots led to a decrease in starch concentration. In roots inoculated in the double sandwich, the polysaccharide accumulated after 1 wk and persisted for at least 4 wk. The response was absent in the castor myc(-) mutant, sym4-2, while transcript levels of both CASTOR and POLLUX were slightly enhanced in the wild-type L. japonicus roots, suggesting a requirement of the corresponding proteins for the starch-accumulation response. Exudates obtained from fungal spores germinated in the absence of the plant also induced starch accumulation in wild-type L. japonicus roots. * We conclude that factors released from germinating AM fungal spores induce changes in the root carbon status, possibly by enhancing sugar import, which leads to starch accumulation when colonization is prevented. PMID- 19555371 TI - A mid-gut microbiota is not required for the pathogenicity of Bacillus thuringiensis to diamondback moth larvae. AB - The mode of action of the entomopathogenic bacterium Bacillus thuringiensis (Bt) remains a matter of debate. Recent reports have claimed that aseptic lepidopteran hosts were not susceptible to Bt and that inoculation with mid-gut bacteria restores pathogenicity. These claims are controversial because larvae were rendered aseptic by consuming antibiotics, although the effect of these antibiotics on Bt was not examined. We tested the generality of the mid-gut bacteria hypothesis in the diamondback moth, Plutella xylostella using properly controlled experiments that investigated the effect of antibiotic consumption and absence of gut microbiota separately. We found that purified Bt toxin and spore/toxin mixtures were fully pathogenic to larvae reared aseptically. Persistence of antibiotics in larval tissues was implicated in reducing host mortality because larval consumption of the antibiotic rifampicin reduced the pathogenicity of rifampicin-sensitive Bt strains but not rifampicin-resistant strains. Inoculating larvae with Enterobacter sp. Mn2 reduced the mortality of larvae feeding on Bt HD-1 and the presence of a culturable gut microbiota also reduced the pathogenicity of the Bt toxin Cry1Ac, in agreement with other studies indicating that an intestinal microbiota can protect taxonomically diverse hosts from pathogen attack. As ingestion of antibiotics suppresses host mortality the vegetative growth of Bt in the host must be important for its pathogenicity. Furthermore, claims that aseptic larvae are not susceptible to Bt must be supported by experiments that control for the effect of administering antibiotics. PMID- 19555370 TI - Gene expression profiling of aging in multiple mouse strains: identification of aging biomarkers and impact of dietary antioxidants. AB - We used DNA microarrays to identify panels of transcriptional markers of aging that are differentially expressed in young (5 month) and old (25 month) mice of multiple inbred strains (129sv, BALB/c, CBA, DBA, B6, C3H and B6C3F(1)). In the heart, age-related changes of five genes were studied throughout the mouse lifespan: complement component 4, chemokine ligand 14, component of Sp100-rs, phenylalanine hydroxylase and src family associated phosphoprotein 2. A similar analysis in the brain (cerebellum) involved complement component 1q (alpha polypeptide), complement component 4, P lysozyme structural, glial fibrillary acidic protein and cathepsin S. Caloric restriction (CR) inhibited age-related expression of these genes in both tissues. Parametric analysis of gene set enrichment identified several biological processes that are induced with aging in multiple mouse strains. We also tested the ability of dietary antioxidants to oppose these transcriptional markers of aging. Lycopene, resveratrol, acetyl-l carnitine and tempol were as effective as CR in the heart, and alpha-lipoic acid and coenzyme Q(10) were as effective as CR in the cerebellum. These findings suggest that transcriptional biomarkers of aging in mice can be used to estimate the efficacy of aging interventions on a tissue-specific basis. PMID- 19555372 TI - Chromosomal antioxidant genes have metal ion-specific roles as determinants of bacterial metal tolerance. AB - Microbiological metal toxicity involves redox reactions between metal species and cellular molecules, and therefore, we hypothesized that antioxidant systems might be chromosomal determinants affecting the susceptibility of bacteria to metal toxicity. Here, survival was quantified in metal ion-exposed planktonic cultures of several Escherichia coli strains, each bearing a mutation in a gene important for redox homeostasis. This characterized approximately 250 gene-metal combinations and identified that sodA, sodB, gor, trxA, gshA, grxA and marR have distinct roles in safeguarding or sensitizing cells to different toxic metal ions (Cr(2)O(7)(2-), Co(2+), Cu(2+), Ag(+), Zn(2+), AsO(2)(-), SeO(3)(2-) or TeO(3)(2 )). To shed light on these observations, fluorescent sensors for reactive oxygen species (ROS) and reduced thiol (RSH) quantification were used to ascertain that different metal ions exert oxidative toxicity through disparate modes-of-action. These oxidative mechanisms of metal toxicity were categorized as involving ROS and thiol-disulfide chemistry together (AsO(2)(-), SeO(3)(2-)), ROS predominantly (Cu(2+), Cr(2)O(7)(2-)) or thiol-disulfide chemistry predominantly (Ag(+), Co(2+), Zn(2+), TeO(3)(2-)). Corresponding to this, promoter-luxCDABE fusions showed that toxic doses of different metal ions up- or downregulate the transcription of gene sets marking distinct pathways of cellular oxidative stress. Altogether, our findings suggest that different metal ions are lethal to cells through discrete pathways of oxidative biochemistry, and moreover, indicate that chromosomally encoded antioxidant systems may have metal ion-specific physiological roles as determinants of bacterial metal tolerance. PMID- 19555373 TI - Metagenomic analysis of viruses in reclaimed water. AB - Reclaimed water use is an important component of sustainable water resource management. However, there are concerns regarding pathogen transport through this alternative water supply. This study characterized the viral community found in reclaimed water and compared it with viruses in potable water. Reclaimed water contained 1000-fold more virus-like particles than potable water, having approximately 10(8) VLPs per millilitre. Metagenomic analyses revealed that most of the viruses in both reclaimed and potable water were novel. Bacteriophages dominated the DNA viral community in both reclaimed and potable water, but reclaimed water had a distinct phage community based on phage family distributions and host representation within each family. Eukaryotic viruses similar to plant pathogens and invertebrate picornaviruses dominated RNA metagenomic libraries. Established human pathogens were not detected in reclaimed water viral metagenomes, which contained a wealth of novel single-stranded DNA and RNA viruses related to plant, animal and insect viruses. Therefore, reclaimed water may play a role in the dissemination of highly stable viruses. Information regarding viruses present in reclaimed water but not in potable water can be used to identify new bioindicators of water quality. Future studies will need to investigate the infectivity and host range of these viruses to evaluate the impacts of reclaimed water use on human and ecosystem health. PMID- 19555374 TI - A digital tongue imaging system for tongue coating evaluation in patients with oral malodour. AB - OBJECTIVE: Several simple visual methods have been developed for assessing tongue coating, but it is difficult to eliminate biases associated with these. The digital tongue imaging system (DTIS) was designed to acquire tongue surface images using a digital camera under controlled conditions, and to calculate tongue coating area. The aim of this study was to evaluate the potential of DTIS for clinical use by comparing it with the Winkel tongue coating index (WTCI). MATERIALS AND METHODS: Forty participants with oral malodour were rated on WTCI by two independent examiners, and photographs of their tongues were assessed using the DTIS. The photographs were also rated by the examiners (blinded to in vivo WTCI and DTIS statuses). RESULTS: Agreements between in vivo WTCI ratings and DTIS assessments were relatively high at r = 0.561 for one examiner and r = 0.736 for the other (P < 0.01), while agreements between the in vivo WTCI ratings and tongue photograph-based ratings were also high at r = 0.645 for one examiner and r = 0.742 (P < 0.01) for the other. CONCLUSIONS: Digital tongue imaging system was found to be highly reliable and as having potential clinical applications. However, the algorithm for determining in vivo tongue coating status requires improvement. PMID- 19555375 TI - Impact of allogeneic haematopoietic stem cell transplantation in the outcome of Ph+ acute lymphoblastic leukaemia treated with an imatinib-containing regimen. PMID- 19555376 TI - Hydrophilic and lipophilic moisturizers have similar penetration profiles but different effects on SC water distribution in vivo. AB - Dry skin is often treated with hydrophilic and/or lipophilic moisturizers. Hydrophilic moisturizers must penetrate the stratum corneum (SC) deeply to function properly, whereas lipophilic moisturizers should remain in the upper SC layers. In this study, both types of moisturizers were applied on volunteers for 3 h, after which the relative amount of moisturizer and the water distribution in the SC were determined using attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy in combination with tape-stripping. The results show that while hydrophilic moisturizers penetrate much more readily than lipophilic moisturizers, the latter are abundantly present in the upper regions of the SC. It was also observed that a 3-h treatment with lipophilic moisturizer did not result in increased water levels in the SC, whereas hydrophilic moisturizers retained water where they are located. The results suggest that upon prolonged application, adequate amounts of moisturizer can be obtained in those regions where they may cause moisturization in the central part of the SC. However, a single application of 3 h is probably too short to exert increased hydration as measured with ATR-FTIR. PMID- 19555377 TI - Patient perspectives on health advice posted on Internet discussion boards: a qualitative study. AB - BACKGROUND: Use of the Internet for health information by patients is growing, and there have been diverse responses to this both within the research community and the medical and health-related professions. The use of Internet discussion boards are one way that people living with long-term conditions can interact with their peers and offer and seek advice, support and information. We report patient perspectives on using a discussion board within a wider pilot study of an Internet-based self-management system for diabetes. DESIGN: Qualitative data was gathered during three stages of developing and piloting the wider self-management system. These are: (1) patient focus groups as part of a stakeholder consultation; (2) a pre-test session and focus group; and 3. a 6-month pilot study including follow-up individual interviews. RESULTS: Three main themes were identified within participants' perspectives on Internet discussion boards. First, a focus on the importance and value of peer support to these patients. Secondly, participants' awareness of the need to evaluate the information posted by others in light of their own circumstances. Thirdly, the value placed upon the experiential knowledge of others living with the same condition. CONCLUSIONS: Many people living with long-term conditions would like to be in contact with their peers, and Internet discussion boards represent a cost-effective and interactive way of achieving this. Within the context of diabetes, the knowledge and expertise accumulated over many years of self-management is central to participants' self-reported ability to evaluate information posted and make decisions on its possible use. PMID- 19555379 TI - Priority effects in experimental populations of the cyanobacterium Microcystis. AB - The arrival order of colonists in developing populations can have a lasting influence on community and population structure, a phenomenon referred to as priority effects. To explore whether such priority effects are important in determining strain composition of populations of the cyanobacterium Microcystis, four Microcystis strains, isolated from a single lake and differing in functional traits, were grown during 4 weeks in the laboratory in all possible pairwise combinations, with the two strains either inoculated at the same time or with a time lag of 1 week, in the presence or absence of grazing Daphnia magna. The relative abundance of strains in the mixtures was assessed using denaturing gradient gel electrophoresis, and the growth rate of each strain in the mixtures was determined for the last 2 weeks of the experiment. We observed strong effects of inoculation order on the final population structure, and these effects were influenced by grazing Daphnia. The priority effects were strain-specific and occurred in two directions: some of the strains grew slower while others grew faster when inoculated second compared with when inoculated first. Our results indicate that priority effects may have a profound impact on strain composition of Microcystis populations. PMID- 19555380 TI - Nodulation of Sesbania species by Rhizobium (Agrobacterium) strain IRBG74 and other rhizobia. AB - Concatenated sequence analysis with 16S rRNA, rpoB and fusA genes identified a bacterial strain (IRBG74) isolated from root nodules of the aquatic legume Sesbania cannabina as a close relative of the plant pathogen Rhizobium radiobacter (syn. Agrobacterium tumefaciens). However, DNA:DNA hybridization with R. radiobacter, R. rubi, R. vitis and R. huautlense gave only 44%, 5%, 8% and 8% similarity respectively, suggesting that IRBG74 is potentially a new species. Additionally, it contained no vir genes and lacked tumour-forming ability, but harboured a sym-plasmid containing nifH and nodA genes similar to those in other Sesbania symbionts. Indeed, IRBG74 effectively nodulated S. cannabina and seven other Sesbania spp. that nodulate with Ensifer (Sinorhizobium)/Rhizobium strains with similar nodA genes to IRBG74, but not species that nodulate with Azorhizobium or Mesorhizobium. Light and electron microscopy revealed that IRBG74 infected Sesbania spp. via lateral root junctions under flooded conditions, but via root hairs under non-flooded conditions. Thus, IRBG74 is the first confirmed legume-nodulating symbiont from the Rhizobium (Agrobacterium) clade. Cross inoculation studies with various Sesbania symbionts showed that S. cannabina could form fully effective symbioses with strains in the genera Rhizobium and Ensifer, only ineffective ones with Azorhizobium strains, and either partially effective (Mesorhizobium huakii) or ineffective (Mesorhizobium plurifarium) symbioses with Mesorhizobium. These data are discussed in terms of the molecular phylogeny of Sesbania and its symbionts. PMID- 19555381 TI - Molecular analysis of the phosphorus starvation response in Trichodesmium spp. AB - The marine diazotroph Trichodesmium is a major contributor to primary production and nitrogen fixation in the tropical and subtropical oceans. These regions are often characterized by low phosphorus (P) concentrations, and P starvation of Trichodesmium could limit growth, and potentially constrain nitrogen fixation. To better understand how this genus responds to P starvation we examined four genes involved in P acquisition: two copies of a high-affinity phosphate binding protein (pstS and sphX) and two putative alkaline phosphatases (phoA and phoX). Sequence analysis of these genes among cultured species of Trichodesmium (T. tenue, T. erythraeum, T. thiebautii and T. spiralis) showed that they all are present and conserved within the genus. In T. erythraeum IMS101, the expression of sphX, phoA and phoX were sensitive to P supply whereas pstS was not. The induction of alkaline phosphatase activity corresponded with phoA and phoX expression, but enzyme activity persisted after the expression of these genes returned to basal levels. Additionally, nifH (nitrogenase reductase; involved in nitrogen fixation) expression was downregulated under P starvation conditions. These data highlight molecular level responses to low P and lay a foundation for better understanding the dynamics of Trichodesmium P physiology in low-P environments. PMID- 19555383 TI - Open or laparoscopic appendicectomy? PMID- 19555382 TI - Hypocretin/orexin and narcolepsy: new basic and clinical insights. AB - Narcolepsy is a chronic sleep disorder, characterized by excessive daytime sleepiness (EDS), cataplexy, sleep paralysis and hypnagogic hallucinations. Both sporadic (95%) and familial (5%) forms of narcolepsy exist in humans. The major pathophysiology of human narcolepsy has been recently discovered based on the discovery of narcolepsy genes in animals; the genes involved in the pathology of the hypocretin/orexin ligand and its receptor. Mutations in hypocretin-related genes are rare in humans, but hypocretin ligand deficiency is found in a large majority of narcolepsy with cataplexy. Hypocretin ligand deficiency in human narcolepsy is probably due to the post-natal cell death of hypocretin neurones. Although a close association between human leucocyte antigen (HLA) and human narcolepsy with cataplexy suggests an involvement of autoimmune mechanisms, this has not yet been proved. Hypocretin deficiency is also found in symptomatic cases of narcolepsy and EDS with various neurological conditions, including immune mediated neurological disorders, such as Guillain-Barre syndrome, MA2-positive paraneoplastic syndrome and neuromyelitis optica (NMO)-related disorder. The findings in symptomatic narcoleptic cases may have significant clinical relevance to the understanding of the mechanisms of hypocretin cell death and choice of treatment option. The discoveries in human cases lead to the establishment of the new diagnostic test of narcolepsy (i.e. low cerebrospinal fluid hypocretin-1 levels for 'narcolepsy with cataplexy' and 'narcolepsy due to medical condition'). As a large majority of human narcolepsy patients are ligand deficient, hypocretin replacement therapy may be a promising new therapeutic option, and animal experiments using gene therapy and cell transplantations are in progress. PMID- 19555384 TI - The impact of lymph node yield on Duke's B and C colorectal cancer survival. AB - AIM: The number of positive lymph nodes retrieved following colorectal cancer (CRC) resection impacts on the staging and further treatment of the disease. We compared 5-year survival by lymph node yield for Duke's B and C patients to assess the impact on prognosis. METHOD: A retrospective methodology was employed to review patients who underwent operative resection for Duke's B or C CRC between 1999 and 2003. RESULTS: A total of 351 patients were included in our analyses. Lymph node yield, N-stage and extramural vascular invasion were independent predictors of overall 5-year survival. A significant difference in 5 year survival by lymph node yield was seen among Duke's B patients (< 9 nodes vs >= 9 nodes, 45.2%vs 68.4%; P = 0.0043) and Duke's C patients (< 10 nodes vs >= 10 nodes, 25.6%vs 48.8%; P = 0.0099). There was a significant reduction in the relative risk of 2.8% in mortality for each additional node sampled in Duke's B and C patients (RR 0.972, 95% confidence interval 0.949-0.994, P = 0.0102). Duke's B patients who had < 9 lymph node yield and no neoadjuvant/adjuvant treatment had a similar survival to all Duke's C patients (47.8%vs 41.7%, P = 0.5136). CONCLUSION: Lymph node yield independently predicts for survival in patients with Duke's B and C CRC. Duke's B patients with < 9 lymph node yield have no better survival than patients with Duke's C disease. Therefore, prospective randomized studies are required to examine if inadequate lymph node yield could be one of the deciding factors in offering adjuvant therapy among Duke's B cancer patients. PMID- 19555385 TI - Severe mesorectal bleeding after stapled transanal rectal resection (STARR operation) using the 'Contour Transtar curved cutter stapler'. PMID- 19555386 TI - Meta-analysis of the colon J-pouch vs transverse coloplasty pouch after anterior resection for rectal cancer. AB - PURPOSE: To evaluate the outcome of colonic J-pouches (CJP) and transverse colonic pouches (TCPs) after anterior resection for rectal cancer. METHOD: Trials were located through Medline, Embase, the Cochrane Central Register of Controlled Trials, VIP and CNKI. Main end-points included functional outcomes, postoperative complications and anorectal physiological outcomes. RESULTS: Of 120 articles, 34 compared CJP and TCP. Of these only six were randomized controlled trials (RCT), which fulfilled the inclusion criteria. These six included 648 patients, including 326 in the CJP group and 322 in the TCP group. There were no differences in the incidences of anastomotic leak [odds ratio 0.50, 95% confidence interval (CI) 0.21-1.18], chest infection (0.43, 0.09-2.00), wound infection (0.87, 0.33-2.30), anastomotic stricture (1.30, 0.44-3.84), fistula (0.64, 0.18-2.31).There were no difference in functional outcomes such as stool frequency [weighted mean difference (WMD) of -0.01, -0.30-0.27 at 6 months].There was no difference for anorectal physiology but heterogeneity existed: resting pressure (0.39, -1.76 to 2.55; 3.09, -0.04 to 6.23; 4.15, 2.21-6.094, at preoperation, 6 and 12 months,); squeeze pressure (-15.02, -46.14 to 16.10; 15.04, -37.04 to 6.97;0.83, -7.70 to 9.37 at preoperation, 6 and 12 months);(Neo)rectal threshold volume(8.49, 5.18-11.81; 27.13, -5.08 to 59.35, at preoperation and 6 months); Maximal (neo) rectal volume (-14.05, -36.60 to 8.50; 23.37, 2.65-44.09; -0.54, -0.91 to -0.18, at preoperation, 6 and at 12 months). CONCLUSIONS: Transverse colonic pouch has similar results as CJP. As it is a safe, feasible, simple, technically easy and time-saving surgical procedure, TCP is a good candidate for wider clinical application. PMID- 19555387 TI - Sentinel node biopsy and localized resection of colon cancer. PMID- 19555388 TI - Complete clinical response to neoadjuvant chemoradiotherapy in patients with rectal cancer: opinions of British and Irish specialists. AB - INTRODUCTION: Advances in neoadjuvant treatment have highlighted the phenomenon of complete clinical response (CCR) in a proportion of patients with rectal cancer. Radical surgery may be associated with a poor functional outcome and quality of life and has a small but significant risk of mortality. This study aimed to assess opinion of colorectal surgeons on issues surrounding the question of nonoperative management in patients who demonstrate complete response after neoadjuvant therapy. METHOD: A questionnaire was sent to members of the Association of Coloproctology of Great Britain and Ireland regarding investigations, clinical management, pathological assessment and oncological outcome in rectal cancer patients with a complete response to neoadjuvant chemoradiotherapy. RESULTS: A total of 122 consultants responded (26% response rate). Most surgeons (58%) would not consider conservative management of patients with a complete response and even more (69%) expressed that they would never discuss nonoperative management in patients with rectal cancer who are fit for curative surgery. Over 70 different combinations of investigations and imaging modalities were suggested to define a CCR. Eighty-six per cent of consultants felt that a pathology report stating no evidence of residual adenocarcinoma did not rule out the presence of tumour cells and all respondents estimated the percentage of patients with pathological complete response as < 20%. CONCLUSIONS: No consensus exists as to what defines a complete response and at present there is resistance to offering nonoperative management in selected patients. With improvements in neoadjuvant treatment modalities, it will be increasingly important to consider nonoperative management in the future. PMID- 19555389 TI - Comparison of the prognostic value of tumour- and patient-related factors in patients undergoing potentially curative surgery for colon cancer. AB - AIM: To comprehensively compare the prognostic value of tumour- and patient related factors in patients undergoing curative surgery for colon cancer. METHOD: From a database of 287 patients who underwent elective resection between 1997 and 2005, tumour factors including stage and host factors including systemic inflammatory response [modified Glasgow Prognostic Score (mGPS)] were identified. RESULTS: Median follow-up was 65 months. Over this period, 125 patients died, 80 from cancer. On multivariate analysis of all significant patient and tumour related factors, Dukes stage (P < 0.01), vascular invasion (P < 0.01) and the mGPS (P < 0.01) were independently associated with cancer-survival. Of the patient-related factors, age (P < 0.01), haemoglobin (P < 0.01), white-cell (P < 0.01), neutrophil (P < 0.01) and platelet (P < 0.01) counts, and alkaline phosphatase (P < 0.01) were most significantly associated with the mGPS. CONCLUSION: In addition to tumour-related factors such as Dukes stage and vascular invasion, the preoperative mGPS should be included to guide prognosis in patients undergoing curative resection for colon cancer. PMID- 19555390 TI - Therapeutic antitumor efficacy of monoclonal antibody against Claudin-4 for pancreatic and ovarian cancers. AB - Claudin-4 (CLDN4) is a tetraspanin transmembrane protein of tight junction structure and is highly expressed in pancreatic and ovarian cancers. In this study, we aimed to generate an anti-Claudin-4 monoclonal antibody (mAb) and evaluate its antitumor efficacy in vitro and in vivo. To isolate specific mAb, we generated CLDN3, 4, 5, 6, and 9, expressing Chinese hamster ovary (CHO) cells, and then used them as positive and negative targets through cell-based screening. As a result, we succeeded in isolating KM3900 (IgG2a), which specifically bound to CLDN4, from BXSB mice immunized with pancreatic cancer cells. Immunoprecipitation and flow cytometry analysis revealed that KM3900 recognized the conformational structure and bound to extracellular loop 2 of CLDN4. Furthermore, binding of KM3900 was detected on CLDN4-expressing pancreatic and ovarian cancer cells, but not on negative cells. Next, we made the mouse-human chimeric IgG1 (KM3934) and evaluated its antitumor efficacy. KM3934 induced dose dependent antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity in vitro, and significantly inhibited tumor growth in MCAS or CFPAC 1 xenograft SCID mice in vivo (P < 0.05). These results suggest that mAb therapy against CLDN4 is promising for pancreatic and ovarian cancers. PMID- 19555391 TI - Mutations in the mitochondrial genome confer resistance of cancer cells to anticancer drugs. AB - The majority of cancer cells harbor homoplasmic somatic mutations in the mitochondrial genome. We show here that mutations in mitochondrial DNA (mtDNA) are responsible for anticancer drug tolerance. We constructed several trans mitochondrial hybrids (cybrids) with mtDNA derived from human pancreas cancer cell lines CFPAC-1 and CAPAN-2 as well as from healthy individuals. These cybrids contained the different mitochondrial genomes with the common nuclear background. We compared the mutant and wild-type cybrids for resistance against an apoptosis inducing reagent and anticancer drugs by exposing the cybrids to staurosporine, 5 fluorouracil, and cisplatin in vitro, and found that all mutant cybrids were more resistant to the apoptosis-inducing and anticancer drugs than wild-type cybrids. Next, we transplanted mutant and wild-type cybrids into nude mice to generate tumors. Tumors derived from mutant cybrids were more resistant than those from wild-type cybrids in suppressing tumor growth and inducing massive apoptosis when 5-fluorouracil and cisplatin were administered. To confirm the tolerance of mutant cybrids to anticancer drugs, we transplanted a mixture of mutant and wild type cybrids at a 1:1 ratio into nude mice and examined the effect by the drugs on the drift of the ratio of mutant and wild-type mtDNA. The mutant mtDNA showed better survival, indicating that mutant cybrids were more resistant to the anticancer drugs. Thus, we propose that mutations in the mitochondrial genome are potential targets for prognosis in the administration of anticancer drugs to cancer patients. PMID- 19555393 TI - Molecular genetics of sarcomas: applications to diagnoses and therapy. AB - Sarcomas are mesenchymal cancers consisting of tumors with various clinical and pathological features. Some of them compel affected individuals to lose important musculoskeletal functions, and some of them are highly malignant and life threatening. A great amount of genetic information for sarcomas has accumulated during the past two decades, contributing diagnoses and treatments. From the standpoint of molecular genetics, sarcomas are classified into two groups: those with defined genetic alterations and those with various genetic alterations. The genetic alterations in the first group include reciprocal translocations resulting in fusion oncoproteins and oncogenic mutations of defined genes such as those of the c-kit gene in gastrointestinal stromal tumors. The function of fusion proteins includes transcription regulator, signal transducer, chromatic remodeling factor, and growth factor, some of which are suitable targets for the molecular therapy. In tumors belonging to the second group, the number of which is far larger than those of the first group, considerable genetic heterogeneity was found even among tumors with same pathological diagnosis. The disruption of the RB and p53 pathways was frequently found, resulting in the dysregulation of cell cycle and the genomic instability. The application of molecular target therapy for tumors in this group requires novel strategies to overcome cross talk between different signal pathways. Recent evidence from in vitro and in vivo experiments has indicated that the cells of origin of sarcomas are tissue stem cells such as mesenchymal stem cells, and the application of stem cell biology holds the promise of novel treatment options. PMID- 19555392 TI - Constitutive phosphorylation of a Rac GAP MgcRacGAP is implicated in v-Src induced transformation of NIH3T3 cells. AB - MgcRacGAP plays critical roles in cell division through regulating Rho family small GTPases. As we previously reported, phosphorylation of MgcRacGAP on serine 387 (S387) is induced by Aurora B kinase at the midbody during cytokinesis, which is a critical step of cytokinesis. Phosphorylation of S387-MgcRacGAP converts it from RacGAP to RhoGAP, leading to completion of cytokinesis. Here we show that MgcRacGAP is prominently phosphorylated on S387 even in the interphase of v-Src transformed NIH3T3 cells in the cytoplasm, but not in the interphase of parental NIH3T3 or H-RasV12-transformed NIH3T3 cells. Interestingly, levels of phosphorylation on S387 (pS387) correlated with soft agar colony-forming abilities of v-Src-transformed NIH3T3 cells. Expression of a phosphorylation mimic mutant MgcRacGAP-S387D enhanced colony formation of v-Src-transformed NIH3T3 cells. Surprisingly, a Rac1 inhibitor but not kinase inhibitors including Aurora B kinase inhibitor specifically inhibited phosphorylation of S387 MgcRacGAP in v-Src-transformed NIH3T3 cells, suggesting the v-Src-induced pathological positive feedback mechanisms towards Rac1 activation using pS387 MgcRacGAP. These results indicated the difference in the mechanisms between v-Src and H-RasV12-induced transformation, and should shed some light on pathological roles of disordered phosphorylation of MgcRacGAP at S387 in v-Src-induced cell transformation. PMID- 19555394 TI - Morphological changes in hind limb muscles elicited by adjuvant-induced arthritis of the rat knee. AB - We investigated qualitative and quantitative changes in rat hind limb muscles caused by complete Freund's adjuvant (CFA)-induced knee joint pain. One week after CFA injection, muscle atrophy was induced only on the CFA-injected side. Wet weight of the rectus femoris (RF) and soleus (SOL) muscles were significantly decreased by 20% and 19%, respectively. The reduction in cross-sectional areas by CFA was similar for fast and slow muscle fibers in the RF (10% vs 15%, respectively) and SOL muscles (16% vs 16%, respectively). At the light microscopic level, pathological changes were not found in the RF muscles on both sides, although the infiltration of mononuclear cells and muscle regeneration were found in the SOL muscles on CFA-injected and contralateral control sides. On the other hand, electron microscopy revealed degenerative changes in the RF and SOL muscles on the CFA-injected side. Interestingly, sarcomere hypercontraction, indicating overexercise, was observed to a limited extent in the SOL muscles on the control side. In conclusions, knee joint pain can trigger the rapid development of muscle atrophy with degenerative changes not only in thigh but also calf muscles. This indicates that early interventions to inhibit joint pain or inflammation may prevent muscle atrophy. PMID- 19555395 TI - Effects of AT1 receptor and beta1 receptor blocking on blood pressure, peripheral hemodynamic and lipid profile in statin-treated hypertensive hypercholesterolemic patients. AB - Recent evidences suggest a relationship between angiotensin 1 (AT1) receptor gene expression and low density lipoprotein cholesterol (LDL-C) plasma level. We enrolled 16 untreated hypertensive hypercholesterolemic patients (57.4 +/- 7 years old) in a randomized, single-blind, cross-over design. All the patients were allocated to treatment with simvastatin 20 mg/day for 2 weeks, then randomly assigned to telmisartan (40-80 mg/day) or bisoprolol (5-10 mg/day). After 4 weeks the antihypertensive drugs have been withdrawn for a wash-out period of 2 weeks when they were treated with simvastatin alone, then they have been allocated to the alternative antihypertensive treatment for four additional weeks. We measured: systolic (SBP) and diastolic BP (DBP), 24-h mean BP (MBP), Baseline and post-ischemia forearm blood flow (FBF) and vascular resistance (FVR), and Lipid profile. After 2 weeks of treatment with Simvastatin, baseline and post-ischemic FBF increased (both P < 0.05), while baseline and post-ischemic FVR decreased (both P < 0.05). Standing DBP and MBP were reduced more after treatment with telmisartan than with bisoprolol (P < 0.05). Basal and post-ischemic FBF were significantly increased (P < 0.05 and P < 0.005, respectively) and basal and post ischemic FVR significantly decreased (both P < 0.005) only after treatment with telmisartan, as well as plasma triglycerides (TG) (P < 0.05). From this preliminary study carried out on hypercholesterolemic hypertensive patients it appears that the association of telmisartan and simvastatin (but not of bisoprolol and simvastatin) could exert positive effects on a large quantity of vascular functionality parameters, just after a short treatment. PMID- 19555396 TI - Collecting patient race/ethnicity and primary language data in ambulatory care settings: a case study in methodology. AB - OBJECTIVE: To collect patient race/ethnicity and language (r/e/l) in an ambulatory care setting. DATA SOURCES/STUDY SETTING: The Palo Alto Medical Foundation (PAMF), December 2006-May 2008. STUDY DESIGN: Three pilot studies: (1) Comparing mail versus telephone versus clinic visit questionnaire distribution; (2) comparing the front desk method (FDM) versus exam room method (ERM) in the clinic visit; and (3) determining resource allocation necessary for data entry. DATA COLLECTION/EXTRACTION METHODS: Studies were planned and executed by PAMF's Quality and Planning division. PRINCIPAL FINDINGS: Collecting r/e/l data during clinic visits elicited the highest response rate. The FDM yielded higher response rate than the ERM. One full-time equivalent is initially necessary for data entry. CONCLUSIONS: Conducting sequential studies can help guide r/e/l collection in a short time frame. PMID- 19555398 TI - Provider monitoring and pay-for-performance when multiple providers affect outcomes: An application to renal dialysis. AB - OBJECTIVE: To characterize the influence of dialysis facilities and nephrologists on resource use and patient outcomes in the dialysis population and to illustrate how such information can be used to inform payment system design. DATA SOURCES: Medicare claims for all hemodialysis patients for whom Medicare was the primary payer in 2004, combined with the Medicare Enrollment Database and the CMS Medical Evidence Form (CMS Form 2728), which is completed at onset of renal replacement therapy. STUDY DESIGN: Resource use (mainly drugs and laboratory tests) per dialysis session and two clinical outcomes (achieving targets for anemia management and dose of dialysis) were modeled at the patient level with random effects for nephrologist and dialysis facility, controlling for patient characteristics. RESULTS: For each measure, both the physician and the facility had significant effects. However, facilities were more influential than physicians, as measured by the standard deviation of the random effects. CONCLUSIONS: The success of tools such as P4P and provider profiling relies upon the identification of providers most able to enhance efficiency and quality. This paper demonstrates a method for determining the extent to which variation in health care costs and quality of care can be attributed to physicians and institutional providers. Because variation in quality and cost attributable to facilities is consistently larger than that attributable to physicians, if provider profiling or financial incentives are targeted to only one type of provider, the facility appears to be the appropriate locus. PMID- 19555397 TI - Medicaid managed care and health care access for adult beneficiaries with disabilities. AB - OBJECTIVE: To evaluate the impact of Medicaid managed care organizations (MCO) on health care access for adults with disabilities (AWDs). DATA SOURCES: Mandatory and voluntary enrollment data for AWDs in Medicaid MCOs in each county were merged with the Medical Expenditure Panel Survey and the Area Resource File for 1996-2004. STUDY DESIGN: I use logit regression and two evaluation perspectives to compare access and preventive care for AWDs in Medicaid MCOs with FFS. From the state's perspective, I compare AWDs in counties with mandatory, voluntary, and no MCOs. From the enrollee's perspective, I compare AWDs who must enroll in an MCO or FFS to those who may choose between them. PRINCIPAL FINDINGS: Mandatory MCO enrollees are 24.9 percent more likely to wait >30 minutes to see a provider, 32 percent more likely to report a problem accessing a specialist, and 10 percent less likely to receive a flu shot within the past year. These differences persist from the state evaluation perspective. CONCLUSIONS: States should not expect a dramatic change in health care access when they implement Medicaid MCOs to deliver care to the adult disabled population. However, continued attention to specialty care access is warranted for mandatory MCO enrollees. PMID- 19555399 TI - The stable isotope ketoisocaproic acid breath test as a measure of hepatic decarboxylation capacity: a quantitative analysis in normal subjects after oral and intravenous administration. AB - BACKGROUND AND AIMS: There is no generally accepted kinetic evaluation method for the stable isotope [(13)C]ketoisocaproic acid (KIC) breath test. Differences found in the results between women and men are contradictory. METHODS: Oral and intravenous breath tests using 1 mg/kg stable isotope-labelled KIC were performed in healthy male and female volunteers. A power exponential function was fitted to the mass spectrometric data of breath (13)CO(2) enrichment, allowing mathematical analysis of time-to-peak-excretion, half-excretion time, percent label recovery and parameters describing the shape of the curve. Body composition was determined using bioelectrical impedance analysis. RESULTS: After oral administration, total label recovery after 3 h was about 22% and was not different between men (n=7) and women (n=8). The time to maximal label excretion was 0.67 +/- 0.12 h in men and 0.9 +/- 0.32 h in women (P=0.028) and the excretion curve showed an initially slower rise in women compared with men. Adjusting for lean body mass or body water abrogated the sex differences. Total label recovery after intravenous administration was about 9%, suggesting that the substrate was rapidly catabolized in the muscle compartment after intravenous administration. CONCLUSIONS: The modified power exponential function described allows standardized estimates of the KIC breath test results. When corrected for body composition, there are no differences in breath test results between men and women. The comparison between oral and intravenous results provides robust evidence that the KIC breath test measures predominantly hepatic and not muscle decarboxylation and is thus a highly specific liver function test. PMID- 19555400 TI - Interleukin-10 promoter polymorphisms in hepatitis C patients with and without Schistosoma mansoni co-infection. AB - BACKGROUND/AIM: Interleukin (IL-10) plays a major role in chronic hepatitis C virus (HCV) disease pathogenesis, in particular in Schistosoma mansoni (S. mansoni) co-infected patients. Given that interindividual variations in IL-10 production are influenced by functional polymorphisms in the IL-10 gene promoter, we determined the frequencies of common (-1082, -819 and -592) IL-10 promoter polymorphisms in chronic HCV patients with and without S. mansoni co-infection and healthy controls, and investigated their association with the degree of histological activity index (HAI) and response to interferon-ribavirin therapy. METHODS: Genomic DNA from 99 patients and 62 healthy controls, born in the same geographical hyperendemic area, was studied by the polymerase chain reaction, followed by restriction enzyme digestion. Sera were assessed for S. mansoni antibodies. RESULTS: The frequencies of IL-10 polymorphisms at positions -1082, 819 and -592 from the transcription start site were comparable between HCV patients and controls, as well as between HCV mono-infected and either S. mansoni co-infected patients or controls. The grade of inflammation and the stage of fibrosis showed no association with IL-10 polymorphisms. The frequencies of S. mansoni co-infection and IL-10 genotypes/haplotypes were insignificantly different between non-responders and responders to combination therapy. No effect of other factors like age, gender, HAI group scores and serum alanine aminotransferase and aspartate aminotransferase levels was observed on response to therapy in our patients. CONCLUSION: Our findings suggest that common IL-10 ( 1082, -819 and -592) genotypes/haplotypes do not influence the degree of HAI and response to combination therapy or susceptibility to HCV infection with and without S. mansoni co-infection. PMID- 19555401 TI - Predictors of nonresponse to lactulose for minimal hepatic encephalopathy in patients with cirrhosis. AB - BACKGROUND/AIMS: Minimal hepatic encephalopathy (MHE) impairs health-related quality of life and predicts overt hepatic encephalopathy (HE) in cirrhotic patients. Lactulose is effective in the treatment of MHE. However, not all patients respond to lactulose. We evaluated predictors of nonresponse to lactulose. PATIENTS AND METHODS: Consecutive 110 cirrhotic patients without HE were evaluated for MHE by psychometry, P300 auditory event-related potential (P300ERP), venous ammonia and critical flicker frequency (CFF). MHE was diagnosed by abnormal psychometry and P300ERP (>2 SD). MHE patients were treated with lactulose for 1 month. Response was defined by normalization of the abnormal test parameters (both psychometric tests and P300ERP). RESULTS: Sixty patients (54.5%) were diagnosed as having MHE: 17/39 (44%) in Child's A, 21/42 (50%) Child's B and 22/29 (76%) in Child's C. There was a significant difference between Child's C's vs Child's A's and B's (P<0.05). Abnormal psychometric tests and abnormal P300ERP were seen in 74 (67%) and 74 (67%) patients respectively. Of 60 patients with MHE, after treatment, psychometry remained abnormal in 22 (36.6%) and P300ERP in 21 (35%) patients. CFF was<38 Hz in 34 (57%) and 11 (18%) patients, respectively, before and after treatment in MHE patients. There was a significant difference between the baseline serum sodium level (134.7+/-2.6 vs 131.1+/-2.2 mmol/L, P=0.001) and the venous ammonia level (76.6+/-20.7 vs 113.4+/-22.8 micromol/L, P=0.001) between responders vs nonresponders. Receiver operating characteristic analysis was performed to identify the cutoff for venous ammonia [cutoff 93.5 mmol/L, area under the curve (AUC) 0.892 (0.814-0.970)] and for the serum sodium level [cutoff 132.5 mmol/L, AUC 0.874 (0.779-0.998)]. Taking a cutoff of 93.5 mmol/L for ammonia patient had a sensitivity of 88.5% and a specificity of 79.4%, respectively, and a cutoff of 132.5 mmol/L for serum sodium patient had a sensitivity of 76.5% and a specificity of 88.5% for nonresponse to lactulose. On univariate analysis and multivariate analysis, serum sodium and venous ammonia were the only two parameters associated with nonresponse to lactulose. CONCLUSION: The prevalence of MHE was 55% and MHE improved in 57% patients with lactulose. Baseline low serum sodium and high venous ammonia were highly predictive of nonresponse to lactulose therapy. PMID- 19555402 TI - Traumatic carotid-cavernous fistula with pontomesencephalic and cervical cord venous drainage presenting as tetraparesis. AB - A 27-year-old male presented with progressive ascending myelopathy leading to tetraparesis. Magnetic resonance imaging of the cervical spine showed dilated perimedullary veins and spinal cord edema. Catheter angiography demonstrated a direct carotid-cavernous fistula (CCF) with prominent pontomesencephalic and perimedullary venous drainage. Successful coil embolization of the fistula was performed with improvement of the patient's symptoms. To our knowledge, no case of a direct CCF with perimedullary drainage has been previously reported. PMID- 19555403 TI - Thrombosis of a drainage vein in developmental venous anomaly (DVA) leading venous infarction: a case report and review of the literature. AB - Developmental venous anomalies (DVAs) are common congenital venous drainage anomalies. Although they typically have a benign clinical course and a low symptomatic rate, thrombosis of a drainage vein may occur, leading to potentially debilitating complications. We report imaging findings of posterior fossa DVA with a thrombosed drainage vein in a patient with nonhemorrhagic cerebellar infarct. We also review the relevant literature on the subject. PMID- 19555405 TI - PICA origin aneurysm diagnosed 27 years after microvascular decompression of cranial nerve VII. AB - The authors report a case of a posterior inferior cerebellar artery origin aneurysm causing brainstem compression and swallowing difficulty. The patient had an ipsilateral microvascular decompression of cranial nerve VII for hemifacial spasm 27 years prior to the discovery of the aneurysm. The aneurysm was successfully treated endovascularly. A discussion of possible etiologies of the aneurysm's formation is presented. PMID- 19555404 TI - Parry-Romberg syndrome and Rasmussen encephalitis: possible association. Clinical and neuroimaging features. AB - Parry-Romberg syndrome (PRS) is a sporadic disease of unknown etiology with typical onset in childhood or in young adults. It is characterized by a slow and progressive atrophy affecting one side of the face, the skin, the subcutaneous tissue, the muscles, the cartilages, and the underlying bony structures. The neurological symptoms usually include focal epilepsy, migraine, and unilateral brain lesions on the same side as the atrophy. A common neuroimaging finding of the syndrome is white matter high signal intensity on brain magnetic resonance (MR) imaging. Rasmussen encephalitis (RE) is a rare and chronic inflammatory disease of the brain that begins in the first decade of life and more rarely in adolescents and adults. It usually involves one hemisphere with focal cortical inflammation. Neurologic symptoms are intractable seizures and progressive hemiplegia. Both PRS and RE are often associated with other inflammatory or autoimmune disorders and only 1 case of both syndromes has been reported in literature. We report the clinical and neuroradiological findings in a 6-year-old boy, presenting with focal hemifacial and arm motor seizures and progressive facial hemiatrophy. Serial MR imaging studies revealed progressive brain hemispheric signal alterations and atrophy. This would thus suggest acoexistence of PRS and RE. PMID- 19555406 TI - Linguistic history of posterior reversible encephalopathy syndrome: mirror of developing knowledge. AB - BACKGROUND: the term posterior reversible encephalopathy syndrome (PRES) was first proposed in 2000. Since then, the acronym PRES has become very popular in imaging and clinical literature as it is short, easy to say and remember, and neatly couples the frequent localization of neuroimaging findings along with the typical outcome of this syndrome. Another possible reason for the popularity of this acronym in clinical circles is the connotation of PRES with (elevated blood) PRESsure, as a majority of cases are believed to be associated with hypertension. However, problems exist with the interpretation and common understanding of PRES, questioning the appropriateness of "P" and "R" in the acronym. The linguistic issues related to the acronym of PRES are interesting. OBJECTIVES: the aim of this work is to analyze the controversies related to the acronym of PRES. RESULTS: in 2006, modifying the meaning of the acronym was suggested, renaming it Potentially Reversible Encephalopathy Syndrome in order to adjust to the cases when posterior involvement is not prominent and emphasize that the reversibility is not spontaneous. This meant the creation of a backronym, where the new phrase is constructed by starting with an existing acronym. CONCLUSION: this new backronym indicates that the original acronym of PRES has become a misnomer. PMID- 19555407 TI - Diffusion tensor imaging following shunt in a patient with hydrocephalus. AB - We report on a patient with hydrocephalus who was evaluated by diffusion tensor imaging (DTI) follow-up study before and after a shunt operation. A 48-year-old male patient and 6 age-matched control subjects were evaluated. The patient presented with hydrocephalus due to hemorrhage caused by the rupture of a right middle cerebral artery bifurcation aneurysm. Three longitudinal DTIs were acquired from the patient (pre-shunt, post-shunt 2 weeks, and post-shunt 8 weeks). The fractional anisotrophy values in the adjacent structures of the lateral ventricle, which were increased before the shunt operation, were decreased after the shunt operation. We think that DTI could be a useful tool for the evaluation of hydrocephalus. PMID- 19555408 TI - A dural arteriovenous fistula of the anterior cranial fossa angiographically mimicking an anterior ethmoidal artery aneurysm. AB - Venous aneurysm or varix at the venous side of the fistula commonly exist in dural arteriovenous fistula (DAVF) of the anterior cranial fossa, which may be initially mistaken with aneurysm on computed tomography and magnetic resonance imaging, but always identified by angiography. We report a very unusual case of anterior cranial fossa DAVF angiographically mimicking an anterior ethmoidal artery aneurysm, which was ultimately corrected by surgery. A 41-year-old male presented with right frontal intraparenchymal hematoma with intraventricular extension. Angiography revealed a vascular lesion adjacent to the anterior fossa mimicking an anterior ethmoidal artery aneurysm, which was surgically proven to be a partially thrombosed venous varix of drainaging vein originated from the cribriform plate. A diagnosis of anterior cranial fossa DAVF was made, and venous varix was excised. Follow-up angiography after the operation revealed complete disappearance of the lesion. Our case illustrates a unique occasion that a proximal venous varix without obvious outflow angiographically in DAVF might be mistaken with an aneurysm. PMID- 19555410 TI - Assimilation of excess ammonium into amino acids and nitrogen translocation in Arabidopsis thaliana--roles of glutamate synthases and carbamoylphosphate synthetase in leaves. AB - This study was aimed at investigating the physiological role of ferredoxin glutamate synthases (EC 1.4.1.7), NADH-glutamate synthase (EC 1.4.1.14) and carbamoylphosphate synthetase (EC 6.3.5.5) in Arabidopsis. Phenotypic analysis revealed a high level of photorespiratory ammonium, glutamine/glutamate and asparagine/aspartate in the GLU1 mutant lacking the major ferredoxin-glutamate synthase, indicating that excess photorespiratory ammonium was detoxified into amino acids for transport out of the veins. Consistent with these results, promoter analysis and in situ hybridization demonstrated that GLU1 and GLU2 were expressed in the mesophyll and phloem companion cell-sieve element complex. However, these phenotypic changes were not detected in the GLU2 mutant defective in the second ferredoxin-glutamate synthase gene. The impairment in primary ammonium assimilation in the GLT mutant under nonphotorespiratory high-CO(2) conditions underlined the importance of NADH-glutamate synthase for amino acid trafficking, given that this gene only accounted for 3% of total glutamate synthase activity. The excess ammonium from either endogenous photorespiration or the exogenous medium was shifted to arginine. The promoter analysis and slight effects on overall arginine synthesis in the T-DNA insertion mutant in the single carbamoylphosphate synthetase large subunit gene indicated that carbamoylphosphate synthetase located in the chloroplasts was not limiting for ammonium assimilation into arginine. The data provided evidence that ferredoxin glutamate synthases, NADH-glutamate synthase and carbamoylphosphate synthetase play specific physiological roles in ammonium assimilation in the mesophyll and phloem for the synthesis and transport of glutamine, glutamate, arginine, and derived amino acids. PMID- 19555409 TI - Transvenous embolization of a symptomatic venous aneurysm developing in the drainage network of a deep cerebral AVM: a case report. AB - We present a case of a symptomatic venous aneurysm that developed in the drainage network of a deep-seated basal ganglia and thalamic arteriovenous malformation. The venous aneurysm was treated selectively with platinum coils using a venous approach. PMID- 19555411 TI - Multiple paternity increases effective population size. PMID- 19555412 TI - In situ population structure and ex situ representation of the endangered Amur tiger. AB - The Amur tiger (Panthera tigris altaica) is a critically endangered felid that suffered a severe demographic contraction in the 1940s. In this study, we sampled 95 individuals collected throughout their native range to investigate questions relative to population genetic structure and demographic history. Additionally, we sampled targeted individuals from the North American ex situ population to assess the genetic representation found in captivity. Population genetic and Bayesian structure analyses clearly identified two populations separated by a development corridor in Russia. Despite their well-documented 20th century decline, we failed to find evidence of a recent population bottleneck, although genetic signatures of a historical contraction were detected. This disparity in signal may be due to several reasons, including historical paucity in population genetic variation associated with postglacial colonization and potential gene flow from a now extirpated Chinese population. Despite conflicting signatures of a bottleneck, our estimates of effective population size (N(e) = 27-35) and N(e)/N ratio (0.07-0.054) were substantially lower than the only other values reported for a wild tiger population. Lastly, the extent and distribution of genetic variation in captive and wild populations were similar, yet gene variants persisted ex situ that were lost in situ. Overall, our results indicate the need to secure ecological connectivity between the two Russian populations to minimize loss of genetic diversity and overall susceptibility to stochastic events, and support a previous study suggesting that the captive population may be a reservoir of gene variants lost in situ. PMID- 19555413 TI - Invasion or invisibility: using genetic and distributional data to investigate the alien or indigenous status of the Atlantic populations of the peracarid isopod, Stenosoma nadejda (Rezig 1989). AB - The peracarid isopod, Stenosoma nadejda (Rezig 1989), until recently considered to be endemic of the Mediterranean region, was first reported in the Atlantic coast of southern Spain in 2001, and in 2006 abundant populations were discovered throughout the southwestern Portuguese coast. This fast expansion was intriguing because, as a direct brooder, this species has limited mechanisms for dispersal, such as rafting on seaweeds. Did S. nadejda recently extend its range into the Atlantic or was it overlooked in the past? We examined the patterns of genetic diversity and population differentiation accordingly by sequencing the cytochrome c oxidase subunit I mitochondrial gene from 75 individuals collected in five locations in Atlantic Iberia and one in the Mediterranean. Our results indicate that the newly discovered Atlantic populations of S. nadejda appear to be old and have long persisted on Atlantic shores rather than being a recent introduction. High levels of genetic diversity and geographic structure were uncovered in what was initially suspected to be an 'invasive' species. Recent changes in population dynamics may have made S. nadejda more conspicuous in the Atlantic shores, or a more comprehensive survey led to the recognition of this species where it was not expected. PMID- 19555414 TI - Pathogen reduction of fresh plasma using riboflavin and ultraviolet light: effects on plasma coagulation proteins. AB - BACKGROUND: Treatment with riboflavin and ultraviolet (UV) light reduces the pathogens present in blood components. This study assessed changes to the coagulation proteins that had occurred during this treatment of fresh plasma units before freezing. STUDY DESIGN AND METHODS: Twenty fresh plasma units (230 +/- 30 mL) were treated by the Mirasol process (CaridianBCT Biotechnologies) and frozen within 8 hours of donation. Plasma units were combined with 35 mL of a 500 micromol/L riboflavin solution in an illumination bag to achieve a final concentration of approximately 60 micromol/L riboflavin. The bag was placed in the Mirasol illuminator and exposed to UV light (6.24 J/mL). Samples were frozen before and after treatment. RESULTS: Recoveries observed were 67.7 +/- 3.9% Factor (F)XI, 68.5 +/- 3.3% FVIII:C, 78.8 +/- 4.5% fibrinogen, 78.9 +/- 4.1% FV, 79.0 +/- 4.2% FVII, 79.0 +/- 8.6% F IX, 79.7 +/- 2.6% FX, and 85.0 +/- 3.7% FII. Von Willebrand factor (VWF) antigen, VWF:ristocetin cofactor, and ADAMTS13 recoveries were 87.0 +/- 7.1, 85.5 +/- 6.6, and 73.3 +/- 15.2%, respectively, while that of protein C was 83.6 +/- 2.6%. A loss of high-molecular-weight VWF multimers was observed in most units. Recoveries for protein S, antithrombin, and plasmin inhibitor were greater than 90%. The mean FVIII:C concentration, after treatment, was 0.76 +/- 0.17 IU/mL. CONCLUSIONS: As with other pathogen reduction technologies, the Mirasol process resulted in some loss of coagulation factor activity. For most Mirasol-treated units and for most of the tested factors this is unlikely to have clinical impact, but trials are required to demonstrate this. PMID- 19555416 TI - Comparison of frozen versus desiccated reference human red blood cells for hemagglutination assays. AB - BACKGROUND: Red blood cells (RBCs) are commonly used fresh or stored in frozen format for identification of patients' antibodies and serologic specificity of such antibodies at reference laboratories. However, maintaining a large pool of fresh RBCs is impossible in a blood-banking environment and blood in frozen format poses a logistic disadvantage in terms of accessibility, maintenance cost, safety, and sample recovery. This study explores an alternative, desiccation storage method for RBCs to provide a reagent that supports greater utilization and flexibility for reference laboratories. STUDY DESIGN AND METHODS: RBCs from five donors were used in the study. RBCs were processed and kept in either frozen or desiccated format. Study variables for either the frozen or the desiccated cells included cell recovery as quantified by cell counts, gross microscopic examination, and hemagglutination assays. RESULTS: The mean percentage of cell recovery for thawed and washed frozen RBCs was 20% versus 50% for rehydrated and washed desiccated RBCs. Microscopic examination of thawed cells from the frozen preparation showed cells with irregular shapes, a sharp contrast when compared with rehydrated cells from the desiccated preparation, where cells are mostly intact, smooth surface, and biconcave in structure. Cells in both preparations performed well in manual agglutination tests. CONCLUSION: Desiccation preservation of RBCs provides a somewhat better RBC recovery and cell structure stability, while maintaining the necessary antigen-antibody reactions for cell surface markers, which will allow desiccated RBCs to be archived in blood collecting and processing reference laboratories. PMID- 19555417 TI - Quantitation of human herpesvirus 8 (HHV-8) antibody in patients transfused with HHV-8-seropositive blood. AB - BACKGROUND: Human herpesvirus 8 (HHV-8) is endemic in Uganda where seroprevalence is approximately 40%. In a previous study, Ugandan patients receiving blood transfusions had multiple serum specimens collected for 6 months after transfusion to monitor for HHV-8 infection. It was observed that several HHV-8 seronegative patients were unexpectedly HHV-8 seropositive after blood transfusion. STUDY DESIGN AND METHODS: This study measured HHV-8 antibody in serially collected serum specimens from 542 patients who received transfusions and evaluated the risk of HHV-8 infection as a function of HHV-8 antibody levels in the donors. RESULTS: HHV-8 antibody was observed in 52% of patients transfused with HHV-8-seropositive blood in amounts that corresponded with their donor's antibody titer and waned within 40 days. Higher levels of passive HHV-8 antibody in patients who received transfusions appeared to be associated with a lower risk of HHV-8 infection. CONCLUSION: The source of transient antibody in patients who received transfusions was determined to be the transfused blood. Donors with higher HHV-8 antibody titers may have been less likely to have infectious virus in the blood. PMID- 19555415 TI - Minority and foreign-born representation among US blood donors: demographics and donation frequency for 2006. AB - BACKGROUND: Historically, minority populations have represented only a small proportion of US blood donors, but recent trends in immigration and potential blood shortages emphasize the need for recruitment strategies to increase minority donations. STUDY DESIGN AND METHODS: Donation data from a network of six US blood centers for 2006 were analyzed. Race/ethnicity, country of birth, and educational attainment data were collected specifically for the study and assessed for their influence on donation behavior. Logistic regression was used to determine independent associations with repeat donors status and annual donation frequency. RESULTS: A total of 1,288,998 donations from 729,068 donors were studied; most donors had data on race/ethnicity (97.1%) and country of birth (93.1%). The proportion of minority donors differed by blood center, with African American donors (16%) most common at the Southeastern blood center and Asian (12%), Hispanic (13%), and foreign-born donors (13%) most common at the Northern California blood center. Minority donors and those born in Mexico or Latin America were younger than white donors. Minority and non-US-born donors were less likely than white and US-born donors to be repeat donors (odds ratio [OR], 0.60 0.78), and most were less likely to give two or more annual donations (OR, 0.82 1.11). CONCLUSION: Minority and Mexico/Latin America-born donors represent a younger and often first-time donor population compared to white and US-born donors, but their annual donation frequency was only slightly lower than white and US-born donors. Increasing the retention and donation frequency of minorities will be important for supplementing the blood supply. PMID- 19555418 TI - Maintenance of in vitro properties of leukoreduced whole blood-derived pooled platelets after a 24-hour interruption of agitation. AB - BACKGROUND: Continuous agitation during platelet concentrate (PC) storage is frequently interrupted during shipping. Studies have evaluated the effects of interrupted agitation in apheresis and single whole blood-derived PCs, but not PC pools. This study evaluated in vitro properties of pooled whole blood-derived platelets (PLTs) after a 24-hour interruption of agitation. STUDY DESIGN AND METHODS: Eleven ABO-identical leukoreduced whole blood-derived PCs (Leukotrap RC PL, Pall), pooled in a transfer container, were equally divided into each of two CLX-HP containers (Acrodose PL, Pall). One pool (test) was held in a shipping container unagitated for 24 hours between Day 2 and Day 3, while the other (control) was continuously agitated. RESULTS: Ten pairs underwent in vitro assays after 5 and 7 days' storage. Pools contained a mean (+/-SD) of 5.0 x 10(11) +/- 0.4 x 10(11) PLTs. Interrupting agitation for 24 hours reduced test pool pH versus control after 5 days' storage (6.77 +/- 0.15 vs. 6.98 +/- 0.06, p = 0.0005). Test and control pH differences were greater after 7 days' storage (6.17 +/- 0.29 vs. 6.65 +/- 0.14, p < 0.0001); 5 of 10 test pool pHs were less than 6.2 (vs. 0 of 10 controls). Other test pool key in vitro variables were reduced compared with controls after 5 days' storage, with greater differences after 7 days. CONCLUSION: After 5 days' storage, pooled leukoreduced whole blood-derived PCs in CLX-HP containers adequately maintained pH and other key in vitro variables after a 24-hour interruption of agitation. After 7 days' storage, 5 of 10 pools did not maintain a pH value of 6.2 or greater while matched continuously agitated units did. PMID- 19555419 TI - Blood salvage use in gynecologic oncology. AB - BACKGROUND: Blood salvage allows for collection and processing of surgical blood loss with the eventual reinfusion of washed red blood cells (RBCs) back to the patient. The use of blood salvage in patients undergoing surgery for malignancy is off-label. Controversy exists as to the risk of potential cancer dissemination resulting from the reinfusion of the processed blood, but no data are available to confirm this risk. Recent studies have demonstrated that filtering the salvaged blood using a leukoreduction filter (LRF) significantly decreases the number of cancer cells in the recovered RBC aliquot in a variety of cancer types. STUDY DESIGN AND METHODS: Patients on the gynecologic oncology service as part of the bloodless surgery program at Englewood Hospital and Medical Center from April 1998 to April 2007 were identified. Three patients that had reinfusion of cell salvage blood (all reinfusions were performed after filtration with a LRF) were studied further with emphasis placed on long-term outcomes. RESULTS: Two of the three patients did not show any evidence of metastases after surgery. The only patient that developed evidence of hematogenous progression had known liver metastases at the time of her initial diagnosis and therefore had hematogenous dissemination before her index surgery. CONCLUSION: In this series of patients undergoing surgery for malignancies on the gynecologic oncology service, blood salvage with LRF was not definitively associated with hematogenous dissemination. Further large controlled studies are needed to demonstrate the clinical safety of the use of blood salvage in this setting. PMID- 19555420 TI - Easy identification of antibodies to high-prevalence Scianna antigens and detection of admixed alloantibodies using soluble recombinant Scianna protein. AB - BACKGROUND: Identification of antibodies against high-prevalence Scianna (Sc; ERMAP) antigens, like Sc1 and Sc5, is difficult and may incur delays in blood procurement and costs. The detection of additional clinically significant alloantibodies is hampered in the presence of anti-Scianna. Soluble recombinant Scianna protein is demonstrated to facilitate antibody diagnostics in both cases. STUDY DESIGN AND METHODS: Soluble recombinant Scianna protein (Sc:1,-2,3, 4,5,6,7) was produced comprising the antigenic extracellular domain fused to a V5 His tag. The protein was isolated from eukaryotic cell culture supernatants of stably transfected HEK293 cells. Seven serum samples with anti-Sc1, anti-Sc2, and anti-Sc5 and 30 serum samples with antibodies to other blood group antigens were evaluated in hemagglutination inhibition assays. Antisera with mixed antibody specificities and autoantibodies were also tested. RESULTS: Soluble Scianna protein inhibited specifically antibodies to the high-prevalence Scianna antigens Sc1 and Sc5. No antibodies were neutralized that were directed to the low prevalence Sc2 antigen or to a large representative set of antigens from other blood group systems. Clinically relevant antibodies could be identified despite being masked by anti-Sc1 and anti-Sc5. A mixture of Scianna and JMH proteins allowed detecting a common antibody despite the presence of antibodies to high prevalence antigens of the Scianna or JMH blood group systems. CONCLUSION: Antibody detection systems comprising soluble recombinant Scianna protein provide an easy single-step method for detection and identification of antibodies to high prevalence Scianna antigens. Reagents with Scianna and other recombinant blood group proteins and mixtures of such proteins would be useful routine reagents in immunohematology. PMID- 19555421 TI - Trehalose inhibited the phagocytosis of refrigerated platelets in vitro via preventing apoptosis. AB - BACKGROUND: The sole reason that platelets (PLTs) are currently stored at room temperature is that refrigeration of PLTs results in their rapid clearance after transfusion. Recent data have suggested that trehalose was an effective cryoprotective reagent for human PLTs. This study evaluated the effect of trehalose on the phagocytosis of refrigerated PLTs. STUDY DESIGN AND METHODS: Phagocytosis of PLTs was evaluated with THP-1 cells. Phosphatidylserine exposure and caspase-3 activity in PLTs were measured with flow cytometry. PLT mitochondrial transmembrane potentials were studied by staining PLTs with JC-1. Expression of Bcl-XL and Bax was determined by Western blot. PLT aggregation was studied in vitro with a PLT aggregation profiler. RESULTS: It was demonstrated that trehalose significantly inhibited the phagocytosis of refrigerated PLTs by showing that the phagocytotic ratio of PLTs refrigerated with trehalose for 9 days was generally comparable with that of PLTs stored at room temperature for 5 days, which was significantly lower than that of PLTs refrigerated without trehalose. Further studies revealed that trehalose could prevent the apoptosis of refrigerated PLTs, which was determined by the phosphatidylserine exposure, caspase-3 activities, mitochondrial transmembrane potentials, and expression of Bcl-XL and Bax. Finally, it was shown that PLTs refrigerated with trehalose for 9 days also maintained their activity to aggregate when exposed to agonists in a standard aggregometry assay. CONCLUSION: Our results suggest that trehalose could inhibit the phagocytosis of refrigerated PLTs in vitro through preventing apoptosis, implying that trehalose is a promising candidate to optimize the refrigeration of human PLTs. PMID- 19555422 TI - Testing platelet mass versus platelet count to guide platelet transfusions in the neonatal intensive care unit. AB - BACKGROUND: Platelet (PLT) transfusions can bestow significant benefits but they also carry risks. This study sought a safe means of reducing PLT transfusions to neonatal intensive care unit (NICU) patients with thrombocytopenia by comparing two transfusion guidelines, one based on PLT count and the other on PLT mass (PLT count times mean PLT volume). STUDY DESIGN AND METHODS: Using a prospective, two centered, before versus after design, PLT transfusion usage and hemorrhagic events were contrasted during a period when PLT count-based transfusion guidelines were in use (Period 1) versus a period when PLT mass-based guidelines were in use (Period 2). RESULTS: No differences were observed between Periods 1 and 2 in NICU admissions, sex, race/ethnicity, percentage of inborn patients, or percentage of patients with a PLT count less than 50 x 10(9) or 51 x 10(9) to 99 x 10(9)/L. In the first period 3.6% of NICU admissions received one or more PLT transfusions. This fell to 1.9% during the second period (p < 0.002). The number of PLT transfusions administered per transfused patient was the same in both periods: 2.0 (1-23) (median [range]) in Period 1 and 2.0 (1-17) in Period 2 (p > 0.40). Significantly fewer PLT transfusions were given in Period 2 for prophylaxis (patient not bleeding; p < 0.001 vs. Period 1). The number given for bleeding did not change between the two periods. In Period 2 no increases were seen in rate of intraventricular hemorrhage (IVH); Grade 3 or 4 IVH; or pulmonary, gastrointestinal, or cutaneous bleeding. CONCLUSIONS: The use of PLT mass-based NICU transfusion guidelines was associated with fewer PLT transfusions and no recognized increase in hemorrhagic problems. PMID- 19555423 TI - Unintentional fall injuries associated with walkers and canes in older adults treated in U.S. emergency departments. AB - OBJECTIVES: To characterize nonfatal, unintentional, fall-related injuries associated with walkers and canes in older adults. DESIGN: Surveillance data of injuries treated in hospital emergency departments (EDs), January 1, 2001, to December 31, 2006. SETTING: The National Electronic Injury Surveillance System All Injury Program, which collects data from a nationally representative stratified probability sample of 66 U.S. hospital EDs. PARTICIPANTS: People aged 65 and older treated in EDs for 3,932 nonfatal unintentional fall injuries and whose records indicated that a cane or a walker was involved in the fall. MEASUREMENTS: Sex, age, whether the fall involved a cane or walker, primary diagnosis, part of the body injured, disposition, and location and circumstances of the fall. RESULTS: An estimated 47,312 older adult fall injuries associated with walking aids were treated annually in U.S. EDs: 87.3% with walkers, 12.3% with canes, and 0.4% with both. Walkers were associated with seven times as many injuries as canes. Women's injury rates exceeded those for men (rate ratios=2.6 for walkers, 1.4 for canes.) The most prevalent injuries were fractures and contusions or abrasions. Approximately one-third of subjects were hospitalized for their injuries. CONCLUSION: Injuries and hospital admissions for falls associated with walking aids were frequent in this highly vulnerable population. The results suggest that more research is needed to improve the design of walking aids. More information also is needed about the circumstances preceding falls, both to better understand the contributing fall risk factors and to develop specific and effective fall prevention strategies. PMID- 19555424 TI - Synergistic effects of telmisartan and simvastatin on endothelial progenitor cells. AB - Circulating endothelial progenitor cells (EPC) contribute to endothelial replenishment. Telmisartan is an angiotensin-receptor blocker with PPARgamma agonistic properties. PPARgamma-agonists and HMG-CoA reductase inhibitors have been shown to enhance EPC number and function. We focused on the effects of telmisartan alone or in combination with simvastatin on EPC. EPC were isolated from healthy human volunteers, cultured and stimulated with telmisartan, simvastatin, or the combination of telmisartan and simvastatin. Telmisartan significantly increased the number of acLDL/lectin double-positive early EPC, the number of colony forming units (EC-CFU) as well as EPC migratory capacity, inhibited TNFalpha-induced EPC apoptosis and reduced glucose-induced oxidative stress. The telmisartan effect was dose-dependent and could be inhibited by GW9662, indicating a PPARgamma-dependent mechanism. The combination of telmisartan and simvastatin led to a significant additive increase in EPC count and function. In wild-type mice, systemic treatment with either telmisartan or simvastatin elevated the number of sca-1/flk-1-positive EPC in bone marrow and peripheral blood, spleen-derived acLDL/lectin double-positive EPC, EPC migration and EC-CFU. Consistent with the in vitro findings, the combination of telmisartan and simvastatin resulted in a further enhancement of EPC counts. Re endothelialization after carotid injury was significantly enhanced by telmisartan, simvastatin and the combination. Telmisartan increases EPC number and function mediated by a PPARgamma-dependent mechanism. This effect is further enhanced by combination with simvastatin, suggesting a synergistic activation of potentially diverse intracellular pathways. PMID- 19555425 TI - Regression and persistence: remodelling in a tissue engineered axial vascular assembly. AB - In later stages of vasculoangiogenesis a vascular network is going through a metamorphosis for optimal perfusion and economy of energy. In this study we make a quantitative approach to phenomena of remodelling in a bioartificial neovascular network and suggest variance of calibre as a parameter of neovascular maturation. For this study, 18 male Lewis rats were subjected to the AV loop operation in combination with a hard porous biogenic matrix and an isolation chamber. The animals were allocated into three groups for different explantation intervals set to 2, 4 and 8 weeks, respectively. Collective attributes like vascular density, percent fractional area and variance of calibre were evaluated for a predefined region of interest (ROI). Late morphogenesis was evaluated by means of scanning electron microscopy. After the fourth week the absolute number of vessels within the ROI decreased (P < 0.03) whereas, on the contrary, the fractional area of all segments increased (P < 0.02). The variance in calibre was significantly increased in the 8-week group (P < 0.05). Lymphatic growth after week 4, early pericyte migration as well as intussusceptive angiogenesis were identified immunohistologically. Phenomena of remodelling were evaluated quantitatively in a neovascular network and variance could be proposed as a parameter of net vascular maturation. PMID- 19555426 TI - Prescribing statins to patients with nonalcoholic fatty liver disease: real cardiovascular benefits outweigh theoretical hepatotoxic risk. AB - Statins are among the most widely prescribed drugs in the western world and play a significant role in reducing cardiovascular risk. However, concern regarding their hepatic safety profile has meant that patients with concurrent liver pathology are often denied such benefits. In this review we consider the evidence for and against the prescription of statins to patients with nonalcoholic fatty liver disease, a group typically associated with high cardiovascular risk. Contrary to current opinion, we find that there is considerable evidence for and little evidence against the prescription of statins to this population and suggest that the guidelines advising against their use in these patients should be reviewed. PMID- 19555427 TI - Characterization of brain cancer stem cells: a mathematical approach. AB - OBJECTIVE: In recent years, support has increased for the notion that a subpopulation of brain tumour cells in possession of properties typically characteristic of stem cells is responsible for initiating and maintaining the tumour. Unravelling details of the brain tumour stem cell (BTSC) hierarchy, as well as interactions of these cells with various therapies, will be essential in the design of optimal treatment strategies. MATERIALS AND METHODS: Motivated by this, we have developed a mathematical model of the BTSC hypothesis that may aid in characterization of brain tumours, as well as in prediction of effective therapeutic strategies, which can be further validated in experimental and clinical studies. At the level of a small number of cells, the model developed herein is stochastic. For larger populations of cancer cells, the model is handled from a deterministic approach. RESULTS AND CONCLUSIONS: In the stochastic regime, importance of a relationship between the likelihoods of two distinct types of symmetric BTSC divisions in determining BTSC survival rates becomes apparent, consequently emphasizing the need for a set of biomarkers that are able to better characterize the BTSC hierarchy. At the large scale, we predict the importance of the aforementioned symmetric division rates in dictating brain tumour composition. Furthermore, we demonstrate possible therapeutic benefits of considering combination treatments of radiotherapy and putative BTSC inhibitors, such as bone morphogenetic proteins, while reinforcing the importance of developing novel treatment strategies that specifically target the BTSC subpopulation. PMID- 19555428 TI - Fullerene nanomaterials inhibit phorbol myristate acetate-induced inflammation. AB - Inflammation is a natural biological response that occurs when vascular tissues are subjected to harmful stimuli. This process may be beneficial to the host during wound healing and infections but can be detrimental if left unchecked. Oxidative stress, the generation of reactive oxygen species, is thought to be one component of this response. Fullerenes can counteract reactive oxygen species due to their potent antioxidant capabilities. Thus, we hypothesized that these molecules may inhibit inflammation. To test this hypothesis we used an in vivo model of phorbol 12-myristate 13-acetate (PMA)-induced inflammation and examined the effects fullerenes have on mitigating this response. We show that PMA-induced inflammation and oedema is dramatically inhibited when fullerenes are given prior to challenge. Thus, fullerene derivatives may be a novel way to blunt certain inflammatory conditions and facilitate faster recovery of damaged tissue. PMID- 19555429 TI - The effect of moesin overexpression on ageing of human dermal microvascular endothelial cells. AB - Senescence of microvascular endothelial cells is known to play an important role in the pathophysiology of vascular diseases related to ageing, but the accurate mechanism or related genes are not known. Moesin, a cytoskeletal protein and the most potent candidate as an ageing-related protein, showed obvious changes in expression when compared before and after ageing. In this study, a lentivirus was used to overexpress moesin in endothelial cells. The expression of cell cycle mediators such as p16, cyclin D1 and cdk4, which can be the markers of ageing, was compared by RNA and was shown to be suppressed in moesin overexpressed endothelial cells. In conclusion, it can be said that the expression of moesin delays senescence of human dermal microvascular endothelial cells and this fundamental discovery can be used as a basis for understanding the mechanism of ageing and age-related diseases. PMID- 19555430 TI - Association of thymic stromal lymphopoietin gene -847C>T polymorphism in generalized vitiligo. AB - Thymic stromal lymphopoietin (TSLP) induces naive CD4+ T cells to produce Th2 cytokines. In addition, to low production of Th2 cytokines, strong Th1 response, which plays an important role in vitiligo development, has been induced by blockade of TSLP or TSLP receptor. This study examined whether a functional TSLP polymorphism was associated with vitiligo. One hundred and sixty Korean patients with vitiligo and 568 healthy Korean individuals were examined for the four SNPs of TSLP gene. Luciferase activity was measured for promoter assay. The genotype and allele frequencies of -847C>T polymorphism were lower in vitiligo patients compared with the controls, whereas those of wild type were higher (P = 0.004, P = 0.017 respectively). None the less, the promoter activity of -847C decreased significantly (P = 0.013) compared with -847T, expecting lower TSLP mRNA levels in the polymorphism. Collectively, C allele at the TSLP -847C>T polymorphism may increase susceptibility to generalized vitiligo through decreasing TSLP mRNA expression levels. PMID- 19555431 TI - A review of genetic disorders of hypopigmentation: lessons learned from the biology of melanocytes. AB - Inherited diseases of pigmentation were among the first traits studied in humans because of their easy recognition. The discovery of genes that regulate melanocytic development and function and the identification of disease-causative mutations have greatly improved our understanding of the molecular basis of pigmentary genodermatoses and their underlying pathogenetic mechanisms. Pigmentation mutants can account for hypo-/amelanosis, with or without altered melanocyte number, resulting in different phenotypes, such as Waardenburg syndrome, piebaldism, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome, oculocutaneous albinism and Griscelli syndrome. In this review, we summarize the basic concepts of melanocyte biology and discuss how molecular defects in melanocyte development and function can result in the development of hypopigmentary hereditary skin diseases. PMID- 19555432 TI - Oral supplementation with Lactobacillus rhamnosus CGMCC 1.3724 prevents development of atopic dermatitis in NC/NgaTnd mice possibly by modulating local production of IFN-gamma. AB - Prevalence of allergies has increased during the last two decades. Alteration of the gut microbiota composition is thought to play a crucial role in development of atopic diseases. Oral administration of probiotics has been reported to treat and/or prevent symptoms of atopic diseases in infants, but the results are still controversial. We investigated the potential efficacy of dietary interventions by a probiotic strain on prevention and treatment of atopic dermatitis (AD) in a human-like AD model, NC/NgaTnd mice by perinatal administration. Pregnant NC/NgaTnd mice were orally treated with the probiotic strain Lactobacillus rhamnosus CGMCC 1.3724 (LPR), which was followed by treatment of pups until 12 weeks of age. LPR-treated mice exhibited significant lower clinical symptoms of dermatitis, reduced scratching frequency, lower levels of plasma total Immunoglobulin E and higher levels of interferon-gamma in skin biopsies, compared with untreated mice. The protective effect was also observed when mice started to be treated at weaning time (5 weeks of age) even with limited supplementation period of 2 weeks. However, treatment of mice with the probiotic starting 1 week after the onset of the disease (8 weeks of age) had limited effects. The usefulness of LPR for primary prevention of AD was supported. PMID- 19555434 TI - New developments in our understanding of acne pathogenesis and treatment. AB - Interest in sebaceous gland physiology and its diseases is rapidly increasing. We provide a summarized update of the current knowledge of the pathobiology of acne vulgaris and new treatment concepts that have emerged in the last 3 years (2005 2008). We have tried to answer questions arising from the exploration of sebaceous gland biology, hormonal factors, hyperkeratinization, role of bacteria, sebum, nutrition, cytokines and toll-like receptors (TLRs). Sebaceous glands play an important role as active participants in the innate immunity of the skin. They produce neuropeptides, excrete antimicrobial peptides and exhibit characteristics of stem cells. Androgens affect sebocytes and infundibular keratinocytes in a complex manner influencing cellular differentiation, proliferation, lipogenesis and comedogenesis. Retention hyperkeratosis in closed comedones and inflammatory papules is attributable to a disorder of terminal keratinocyte differentiation. Propionibacterium acnes, by acting on TLR-2, may stimulate the secretion of cytokines, such as interleukin (IL)-6 and IL-8 by follicular keratinocytes and IL 8 and -12 in macrophages, giving rise to inflammation. Certain P. acnes species may induce an immunological reaction by stimulating the production of sebocyte and keratinocyte antimicrobial peptides, which play an important role in the innate immunity of the follicle. Qualitative changes of sebum lipids induce alteration of keratinocyte differentiation and induce IL-1 secretion, contributing to the development of follicular hyperkeratosis. High glycemic load food and milk may induce increased tissue levels of 5alpha-dihydrotestosterone. These new aspects of acne pathogenesis lead to the considerations of possible customized therapeutic regimens. Current research is expected to lead to innovative treatments in the near future. PMID- 19555433 TI - p63 directly induces expression of Alox12, a regulator of epidermal barrier formation. AB - Epidermal development and differentiation are tightly controlled processes that culminate in the formation of the epidermal barrier. A critical regulator of different stages of epidermal development and differentiation is the transcription factor p63. More specifically, we previously demonstrated elsewhere that p63 is required for both the commitment to stratification and the commitment to terminal differentiation. We now demonstrate that DeltaNp63alpha, the predominantly expressed p63 isoform in postnatal epidermis, also plays a role in the final stages of epidermal differentiation, namely the formation of the epidermal barrier. We found that DeltaNp63alpha contributes to epidermal barrier formation by directly inducing expression of ALOX12, a lipoxygenase which contributes to epidermal barrier function. Our data demonstrate that DeltaNp63alpha directly interacts with the promoter of Alox12 in the developing epidermis. Furthermore, we found that the induction of Alox12 expression by DeltaNp63alpha depends on intact p63 binding sites in the Alox12 promoter. Finally, we found that DeltaNp63alpha can induce Alox12 expression only in differentiating keratinocytes, consistent with the role of ALOX12 in epidermal barrier formation. PMID- 19555435 TI - Gene duplication and evolutionary novelty in plants. AB - Duplication is a prominent feature of plant genomic architecture. This has led many researchers to speculate that gene duplication may have played an important role in the evolution of phenotypic novelty within plants. Until recently, however, it was difficult to make this connection. We are now beginning to understand how duplication has contributed to adaptive evolution in plants. In this review we introduce the sources of gene duplication and predictions of the various fates of duplicates. We also highlight several recent and pertinent examples from the literature. These examples demonstrate the importance of the functional characteristics of genes and the source of duplication in influencing evolutionary outcome. PMID- 19555436 TI - Cloning and characterization of small RNAs from Medicago truncatula reveals four novel legume-specific microRNA families. AB - MicroRNAs (miRNAs) and small-interfering RNAs (siRNAs) have emerged as important regulators of gene expression in higher eukaryotes. Recent studies indicate that genomes in higher plants encode lineage-specific and species-specific miRNAs in addition to the well-conserved miRNAs. Leguminous plants are grown throughout the world for food and forage production. To date the lack of genomic sequence data has prevented systematic examination of small RNAs in leguminous plants. Medicago truncatula, a diploid plant with a near-completely sequenced genome has recently emerged as an important model legume. We sequenced a small RNA library generated from M. truncatula to identify not only conserved miRNAs but also novel small RNAs, if any. Eight novel small RNAs were identified, of which four (miR1507, miR2118, miR2119 and miR2199) are annotated as legume-specific miRNAs because these are conserved in related legumes. Three novel transcripts encoding TIR-NBS LRR proteins are validated as targets for one of the novel miRNA, miR2118. Small RNA sequence analysis coupled with the small RNA blot analysis, confirmed the expression of around 20 conserved miRNA families in M. truncatula. Fifteen transcripts have been validated as targets for conserved miRNAs. We also characterized Tas3-siRNA biogenesis in M. truncatula and validated three auxin response factor (ARF) transcripts that are targeted by tasiRNAs. These findings indicate that M. truncatula and possibly other related legumes have complex mechanisms of gene regulation involving specific and common small RNAs operating post-transcriptionally. PMID- 19555437 TI - Evaluating glutathione S-transferase (GST) null genotypes (GSTT1 and GSTM1) as a potential biomarker of predisposition for developing leukopenia. AB - Glutathione S-transferase (GST) enzymes protect cells against xenobiotics and oxidative stress products through an electrophilic conjugation process. We investigated the theta (GSTT1) and mu (GSTM1) null genotypes in a group of leukopenic subjects and normal subjects from Northeast Brazil, evaluating their use as biomarkers of susceptibility for developing leukopenia. In a sample-based case-control study, we analysed white blood cell (WBC) counts and GSTT1 and GSTM1 genotypes. A total of 278 subjects were analysed: 91 with leukopenia and 187 controls. GSTT1 null genotype conferred a 5.92-fold risk for occurrence of leukopenia [odds ratios (OR) = 5.92, CI(MLE): 1.64-26.72, P(MLE) = 0.002] and a 3.90-fold risk of neutropenia (OR = 3.90; CI(MLE): 1.05-13.66; P(MLE) = 0.02), while GSTM1 null genotype conferred a 1.78-fold risk for leukopenia (OR = 1.75; CI(MLE): 1.04-3.06, P(MLE) = 0.017) and no risk of neutropenia (OR = 1.71; CI(MLE): 0.88-3.35; P(MLE) = 0.06). The GSTT1, but not the GSTM1 null genotype, was found to be associated with leukopenia and neutropenia. More cellular and molecular studies are needed to evaluate the existence of genotype interactions, and to confirm the appropriateness of using the GSTT1 and/or GSTM1 null genotypes as biomarkers of susceptibility to white blood-cell deficiencies. PMID- 19555438 TI - BAALC and ERG expression in acute myeloid leukemia with normal karyotype: impact on prognosis. AB - Cytogenetic aberrations are important prognostic factors in acute myeloid leukemia (AML). About 45% of de novo AML lack cytogenetic abnormalities, so identification of predictive molecular markers might improve therapy. We studied the prognostic impact of brain and acute leukemia, cytoplasmic (BAALC) and ETS related gene (ERG) expression in AML with normal karyotype. Pretreatment bone marrow samples from 30 cytogenetically normal AML patients were analysed for BAALC and ERG expression using real time RT-PCR. The patients were dichotomized at BAALC and ERG mean expression into low and high expression. BAALC showed high expression in 70% of patients and its expression did not correlate with the clinical parameters of patients. ERG was high in 33.3% of patients and its expression was associated with lower ages and higher white cell counts. With follow-up for 2 years, patients with high BAALC and high ERG had low rates of clinical remission (P < 0.005) and inferior overall survival (OS) (P < 0.001 and <0.002 for BAALC and ERG respectively). No significant association was observed between the increase in BAALC and ERG expression (P = 0.398). Multivariable analysis confirmed high BAALC expression as an independent risk factor for OS. Overexpression of BAALC and ERG either separate or concomitant predict adverse clinical outcome and may define important risk factor in cytogenetically normal AML. PMID- 19555439 TI - An unusual cause of liver masses. PMID- 19555440 TI - Hearing impairment in the Sturge-Weber syndrome. PMID- 19555442 TI - Male personality, life-history strategies and reproductive success in a promiscuous mammal. AB - Recent theoretical work suggests that personality is a component of life history, but links between personality and either age-dependent reproductive success or life-history strategy are yet to be established. Using quantitative genetic analyses on a long-term pedigree we estimated indices of boldness and docility for 105 bighorn sheep rams (Ovis canadensis), born between 1983 and 1999, and compared these indices to their reproductive history from 2 years of age until death. Docility and boldness were highly heritable and negatively genetically correlated. Docile and bold rams survived longer than indocile and shy rams. Docility and boldness had a weak negative effect on reproductive success early in life, but a strong positive effect on older rams. Our findings highlight an important role of personality on reproductive success and suggest that personality could be an important component of life-history strategy. PMID- 19555441 TI - Life cycle abbreviation in trematode parasites and the developmental time hypothesis: is the clock ticking? AB - The typical multi-host life cycle of many parasites, although conferring several advantages, presents the parasites with a highly hazardous transmission route. As a consequence, parasites have evolved various adaptations increasing their chances of transmission between the different hosts of the life cycle. Some trematode species like the opecoelid Coitocaecum parvum have adopted a more drastic alternative strategy whereby the definitive host is facultatively dropped from the cycle, resulting in a shorter, hence easier to complete, life cycle. Like other species capable of abbreviating their life cycle, C. parvum does so through progenetic development within its intermediate host. Laboratory-reared C. parvum can modulate their developmental strategy inside the second intermediate host according to current transmission opportunities, though this ability is not apparent in natural C. parvum populations. Here we show that this difference is likely due to the time C. parvum individuals spend in their intermediate hosts in the natural environment. Although transmission opportunities, i.e. chemical cues of the presence of definitive hosts, promoted the adoption of a truncated life cycle in the early stages of infection, individuals that remained in their amphipod host for a relatively long time had a similar probability of adopting progenesis and the abbreviated cycle, regardless of the presence or absence of chemical cues from the predator definitive host. These results support the developmental time hypothesis which states that parasites capable of facultative life cycle abbreviation should eventually adopt progenesis regardless of transmission opportunities, and provide further evidence of the adaptive plasticity of parasite transmission strategies. PMID- 19555443 TI - Dietary flavonoids enhance conspicuousness of a melanin-based trait in male blackcaps but not of the female homologous trait or of sexually monochromatic traits. AB - Signalling theory predicts that signals should fulfil three fundamental requirements: high detectability, discriminability and, most importantly, reliability. Melanins are the most common pigments in animals. Correlations between genotypic and phenotypic qualities of the sender and size and morph of melanin-based traits are known, but it is contentious whether melanin-based colouration may signal any quality. We examined the effect of supplementing blackcaps (Sylvia atricapilla) with flavonoids, potent plant antioxidants, on plumage colouration. We demonstrate that melanin-based colour can fulfil all requirements of signals of phenotypic condition. As predicted by sexual selection theory, flavonoid supplementation influenced only the sexually dichromatic black cap of males, whereas the female homologous trait and the sexually monochromatic back colouration remained unaffected. Using avian vision models we show that birds can estimate male flavonoid intake from colouration of males' black cap. Because flavonoid ingestion can increase immune responsiveness in blackcaps, melanin head colouration may signal environmentally determined immune condition. PMID- 19555444 TI - Ancestral state reconstruction analysis of hymenopteran sex determination mechanisms. AB - We provide the first phylogenetic evidence supporting complementary sex determination (CSD) as the ancestral mechanism for haplodiploidy in the Hymenoptera. It is currently not possible, however, to distinguish the evolutionary polarity of single locus (sl) CSD and multiple-locus (ml) CSD given the available data. In this light, we discuss the seemingly maladaptive hypothesis of ml-CSD ancestry, suggesting that collapse from ml-CSD to sl-CSD should remain a viable evolutionary hypothesis based on (i) likely weakening of frequency-dependent selection on sex alleles under ml-CSD and (ii) recent findings with respect to the evolutionary novelty of the complementary sex determiner gene in honeybees. Our findings help provide a phylogenetically informed blueprint for future sampling of sex determination mechanisms in the Hymenoptera, as they yield hypotheses for many unsampled or ambiguous taxa and highlight taxa whose further sampling will influence reconstruction of the evolutionary polarity of sex determination mechanisms in major clades. PMID- 19555445 TI - Strategy for increasing detection rates of drug and alcohol abuse in paediatric emergency departments. AB - AIM: To determine whether implementation of criteria for performing a toxicology screen and increasing staff awareness improve detection of substance abuse among adolescents presenting to the emergency department. METHODS: Patients 12 to 18 years of age presenting to one of three emergency departments in Israel were included in a prospective cohort study. In the 'study' hospital, a set of criteria for urine toxicology screen and measurements of ethanol serum level were implemented. No specific interventions were implemented in the two other hospitals. The main outcome measure was the rate of substance abuse detection. RESULTS: The number of adolescents seen in the participating centres was 3200 at the study hospital, and 3493 and 2792 at the two other hospitals. High blood ethanol concentrations were found in 49 patients at the study hospital compared with 30 and 19 patients at the two other hospitals (p < 0.001). Illicit drugs were detected in 13, 4 and 1 patients, respectively (p = 0.002). CONCLUSIONS: Introducing structured guidelines for ordering toxicological screening increases the detection of alcohol and drug of abuse among adolescents presenting to paediatric emergency departments. PMID- 19555446 TI - The effect of prebiotics in the management of neonatal hyperbilirubinaemia. AB - BACKGROUND: Breast milk oligosaccharides such as galacto-oligosaccharides (scGOS) and fructo-oligosaccharides (lcFOS) can influence the intestinal microbial flora. The latter, in turn, can modulate several intestinal and extraintestinal functions, including bilirubin metabolism. Supplementing infant formula with a prebiotic mixture might then be a novel and safe intervention to manage mild neonatal hyperbilirubinaemia. AIM: To investigate the effect of dietary supplementation with prebiotics on moderate hyperbilirubinaemia in healthy, term infants. METHODS: A prospective, double-blind, clinical trial was performed on seventy-six consecutive newborns who were randomly assigned to receive a formula containing 0.8 g/dL of a mixture from scGOS and lcFOS (ratio 9:1), or maltodextrines as placebo for 28 days. Bilirubin levels were determined by the transcutaneous bilirubin measurement within 2 h after birth (T1), at 24, 48 and 72 h and at 5, 7, 10 and 28 days of life. The number of stool per day was also recorded. RESULTS: Neonates receiving prebiotics showed a larger number of stools over all the duration of dietary intervention compared to that of those on placebo (Repeated Measures ANOVA p < 0.001; day 28 3.4 +/- 0.0.9 vs 1.7 +/- 0.9, respectively; Dunn test p < 0.05). Neonates whose formula was supplemented with prebiotics showed a lower transcutaneous bilirubin that was statistically significant from 72 h of life (5.46 +/- 1.6 vs 7.07 +/- 2.49, post hoc Dunn test, p < 0.05) throughout the duration of the dietary intervention (day 28 2.41 +/- 0.4 vs 2.85 +/- 0.5, post hoc Dunn test, p < 0.05). CONCLUSION: The addition of prebiotics to standard infant diet might represent a novel strategy to help control neonatal hyperbilirubinaemia. PMID- 19555447 TI - The effect of rapid diagnostic testing for influenza on the reduction of antibiotic use in paediatric emergency department. AB - AIM: To determine the influence of rapid diagnosis of influenza on antibiotic prescribing to children presenting with influenza-like illness in the emergency department in a inner city hospital in Istanbul, Turkey. METHODS: Patients aged 3 to 14 years presenting to an urban children's teaching hospital emergency department were screened for fever and cough, coryza, myalgias and/or malaise. After obtaining informed consent, patients were allocated into two groups. Group 1: patients were prescribed antibiotics after only physical examination; or Group 2: patients were prescribed antibiotics after rapid influenza testing. Nasopharyngeal swabs obtained from all patients were immediately tested in a single-blind manner with Influenza A/B Rapid Test(R) for influenza A and B. RESULTS: A total of 97 patients were enrolled, and 33 (34%) of these tested positive for influenza. Although frequency of positive results for influenza between the groups was similar (36% vs 32%, respectively), patients in Group 2 were less likely to be prescribed antibiotics when compared to those in Group 1 (32% vs 100%, respectively, p < 0.0001). CONCLUSION: Rapid diagnosis of influenza in the paediatric emergency department may allow a significant reduction in the over-prescription of antibiotics. PMID- 19555448 TI - Melatonin exerts differential actions on X-ray radiation-induced apoptosis in normal mice splenocytes and Jurkat leukemia cells. AB - The ability of melatonin as a potent antioxidant was used as a rationale for testing its antiapoptotic ability in normal cells. Recently, melatonin was shown to possess proapoptotic action by increasing reactive oxygen species in certain cancer cells. The modification of radiation-induced apoptosis by melatonin and the expression of apoptosis-associated upstream regulators were studied in normal mice splenocytes and Jurkat T leukemia cells. C57BL/6 mice were exposed to a single whole body X-ray radiation dose of 2 Gy with or without 250 mg/kg melatonin pretreatment. The Jurkat cells were divided into four groups of control, 1 mm melatonin alone, 4 Gy irradiation-only and melatonin pretreatment before irradiation. The highest level of apoptosis in the normal splenic white pulp was detected by TUNEL assay at 8 hr after irradiation. At this time, the apoptotic index of irradiation-only and melatonin pretreatment groups were 35.6% and 20.7%, respectively. This reduced apoptosis by melatonin was associated with the increase of Bcl-2 expression and a reduction of Bax/Bcl-2 ratio through a relative decrease of p53 mRNA and protein. In the Jurkat cells treated with a combination of melatonin and radiation, both Annexin V-FITC(+)/PI(-) and Annexin V-FITC(+) cells were increased at 48 hr after irradiation when compared with irradiation-only or melatonin alone. The expressions of p53 between groups were well correlated with the results of Annexin V binding. The irradiation or melatonin did not influence the JNK1 expression in Jurkat cells. The present results suggest that melatonin enhances radiation-induced apoptosis in Jurkat leukemia cells, while reducing radiation-induced apoptosis in normal mice splenocytes. These differential effects on radiation-induced apoptosis by melatonin might involve the regulation of p53 expression. PMID- 19555449 TI - Indirect blue light does not suppress nocturnal salivary melatonin in humans in an automobile setting. AB - In 2007, the International Agency for Research on Cancer (IARC) classified shift work that involves circadian disruption as being probably carcinogenic to humans (Group 2A). In this context, light exposure during the night plays a key role because it can suppress nocturnal melatonin levels when exposures exceed a certain threshold. Blue light around 464 nm is most effective in suppressing melatonin because of the spectral sensitivity of melanopsin, a recently discovered photopigment in retinal ganglion cells; the axons of these cells project to the suprachiasmatic nucleus, a circadian master clock in the brain. Due to advances in light technologies, normal tungsten light bulbs are being replaced by light-emitting diodes which produce quasi-monochromatic or white light. The objective of this study was to assess whether the light-melanopsin melatonin axis might be affected in automobiles at night which employ the new generation diodes. To this end, we have tested in an experimental automobile setting whether indirect blue light (lambda(max) = 465 nm) at an intensity of 0.22 or 1.25 lx can suppress salivary melatonin levels in 12 male volunteers (age range 17-27 years) who served as their own controls. Daytime levels were low (2.7 +/- 0.5 pg/mL), and night-time levels without light exposure were high (14.5 +/- 1.1 pg/mL), as expected. Low-intensity light exposures had no significant effect on melatonin levels (0.22 lx: 17.2 +/- 2.8 pg/mL; P > 0.05; 1.25 lx: 12.6 +/- 2.0 pg/mL; P > 0.05). It is concluded that indirect blue light exposures in automobiles up to 1.25 lx do not cause unintentional chronodisruption via melatonin suppression. PMID- 19555450 TI - Evolutionary history shapes the association between developmental instability and population-level genetic variation in three-spined sticklebacks. AB - Developmental instability (DI) is the sensitivity of a developing trait to random noise and can be measured by degrees of directionally random asymmetry [fluctuating asymmetry (FA)]. FA has been shown to increase with loss of genetic variation and inbreeding as measures of genetic stress, but associations vary among studies. Directional selection and evolutionary change of traits have been hypothesized to increase the average levels of FA of these traits and to increase the association strength between FA and population-level genetic variation. We test these two hypotheses in three-spined stickleback (Gasterosteus aculeatus L.) populations that recently colonized the freshwater habitat. Some traits, like lateral bone plates, length of the pelvic spine, frontal gill rakers and eye size, evolved in response to selection regimes during colonization. Other traits, like distal gill rakers and number of pelvic fin rays, did not show such phenotypic shifts. Contrary to a priori predictions, average FA did not systematically increase in traits that were under presumed directional selection, and the increases observed in a few traits were likely to be attributable to other factors. However, traits under directional selection did show a weak but significantly stronger negative association between FA and selectively neutral genetic variation at the population level compared with the traits that did not show an evolutionary change during colonization. These results support our second prediction, providing evidence that selection history can shape associations between DI and population-level genetic variation at neutral markers, which potentially reflect genetic stress. We argue that this might explain at least some of the observed heterogeneities in the patterns of asymmetry. PMID- 19555451 TI - Suppression of constitutive SOS expression by recA4162 (I298V) and recA4164 (L126V) requires UvrD and RecX in Escherichia coli K-12. AB - Sensing DNA damage and initiation of genetic responses to repair DNA damage are critical to cell survival. In Escherichia coli, RecA polymerizes on ssDNA produced by DNA damage creating a RecA-DNA filament that interacts with the LexA repressor inducing the SOS response. RecA filament stability is negatively modulated by RecX and UvrD. recA730 (E38K) and recA4142 (F217Y) constitutively express the SOS response. recA4162 (I298V) and recA4164 (L126V) are intragenic suppressors of the constitutive SOS phenotype of recA730. Herein, it is shown that these suppressors are not allele specific and can suppress SOS(C) expression of recA730 and recA4142 in cis and in trans. recA4162 and recA4164 single mutants (and the recA730 and recA4142 derivatives) are Rec(+), UV(R) and are able to induce the SOS response after UV treatment like wild-type. UvrD and RecX are required for the suppression in two (recA730,4164 and recA4142,4162) of the four double mutants tested. To explain the data, one model suggests that recA(C) alleles promote SOS(C) expression by mimicking RecA filament structures that induce SOS and the suppressor alleles mimic RecA filament at end of SOS. UvrD and RecX are attracted to these latter structures to help dismantle or destabilize the RecA filament. PMID- 19555452 TI - Identification of small RNAs in Mycobacterium tuberculosis. AB - In spite of being one of our most prominent bacterial pathogens, the presence of small regulatory RNAs (sRNAs) has not previously been investigated in Mycobacterium tuberculosis. Post-transcriptional regulation of gene expression by sRNA molecules has been demonstrated in a wide range of pathogenic bacteria and has been shown to play a significant role in the control of virulence. By screening cDNA libraries prepared from low-molecular weight RNA from M. tuberculosis we have identified nine putative sRNA molecules, including cis encoded antisense transcripts from within open reading frames and trans-encoded transcripts from intergenic regions. sRNAs displayed differential expression between exponential and stationary phase, and during a variety of stress conditions. Two of the cis-encoded sRNAs were associated with genes encoding enzymes involved in lipid metabolism, desA1 and pks12. These sRNAs showed complementarity to multiple M. tuberculosis genes, suggesting the potential to act as both cis-encoded and trans-encoded sRNAs. Overexpression of selected trans encoded sRNAs had profound impact on growth of M. tuberculosis and M. smegmatis. This is the first experimental evidence of sRNAs in M. tuberculosis and it will be important to consider the potential influence of sRNA regulation when studying the transcriptome and the proteome of M. tuberculosis during infection. PMID- 19555453 TI - In vivo localizations of membrane stress controllers PspA and PspG in Escherichia coli. AB - The phage shock protein (Psp) response in Gram-negative bacteria counteracts membrane stress. Transcription of the PspF regulon (pspABCDE and pspG) in Escherichia coli is induced upon stresses that dissipate the proton motive force (pmf). Using GFP fusions we have visualized the subcellular localizations of PspA (a negative regulator and effector of Psp) and PspG (an effector of Psp). It has previously been proposed that PspA evenly coates the inner membrane of the cell. We now demonstrate that instead of uniformly covering the entire cell, PspA (and PspG) is highly organized into what appear to be distinct functional classes (complexes at the cell pole and the lateral cell wall). Real-time observations revealed lateral PspA and PspG complexes are highly mobile, but absent in cells lacking MreB. Without the MreB cytoskeleton, induction of the Psp response is still observed, yet these cells fail to maintain pmf under stress conditions. The two spatial subspecies therefore appear to be dynamically and functionally distinct with the polar clusters being associated with sensory function and the mobile complexes with maintenance of pmf. PMID- 19555454 TI - Mycobacteriophage Lysin B is a novel mycolylarabinogalactan esterase. AB - Mycobacteriophages encounter a unique problem among phages of Gram-positive bacteria, in that lysis must not only degrade the peptidoglycan layer but also circumvent a mycolic acid-rich outer membrane covalently attached to the arabinogalactan-peptidoglycan complex. Mycobacteriophages accomplish this by producing two lysis enzymes, Lysin A (LysA) that hydrolyses peptidoglycan, and Lysin B (LysB), a novel mycolylarabinogalactan esterase, that cleaves the mycolylarabinogalactan bond to release free mycolic acids. The D29 LysB structure shows an alpha/beta hydrolase organization with a catalytic triad common to cutinases, but which contains an additional four-helix domain implicated in the binding of lipid substrates. Whereas LysA is essential for mycobacterial lysis, a Giles DeltalysB mutant mycobacteriophage is viable, but defective in the normal timing, progression and completion of host cell lysis. We propose that LysB facilitates lysis by compromising the integrity of the mycobacterial outer membrane linkage to the arabinogalactan-peptidoglycan layer. PMID- 19555455 TI - Listeria monocytogenes L-forms respond to cell wall deficiency by modifying gene expression and the mode of division. AB - Cell wall-deficient bacteria referred to as L-forms have lost the ability to maintain or build a rigid peptidoglycan envelope. We have generated stable, non reverting L-form variants of the Gram-positive pathogen Listeria monocytogenes, and studied the cellular and molecular changes associated with this transition. Stable L-form cells can occur as small protoplast-like vesicles and as multinucleated, large bodies. They have lost the thick, multilayered murein sacculus and are surrounded by a cytoplasmic membrane only, although peptidoglycan precursors are still produced. While they lack murein-associated molecules including Internalin A, membrane-anchored proteins such as Internalin B are retained. Surprisingly, L-forms were found to be able to divide and propagate indefinitely without a wall. Time-lapse microscopy of fluorescently labelled L forms indicated a switch to a novel form of cell division, where genome containing membrane vesicles are first formed within enlarged L-forms, and subsequently released by collapse of the mother cell. Array-based transcriptomics of parent and L-form cells revealed manifold differences in expression of genes associated with morphological and physiological functions. The L-forms feature downregulated metabolic functions correlating with the dramatic shift in surface to volume ratio, whereas upregulation of stress genes reflects the difficulties in adapting to this unusual, cell wall-deficient lifestyle. PMID- 19555457 TI - Unexpected chemoreceptors mediate energy taxis towards electron acceptors in Shewanella oneidensis. AB - Shewanella oneidensis uses a wide range of terminal electron acceptors for respiration. In this study, we show that the chemotactic response of S. oneidensis to anaerobic electron acceptors requires functional electron transport systems. Deletion of the genes encoding dimethyl sulphoxide and trimethylamine N oxide reductases, or inactivation of these molybdoenzymes as well as nitrate reductase by addition of tungstate, abolished electron acceptor taxis. Moreover, addition of nigericin prevented taxis towards trimethylamine N-oxide, dimethyl sulphoxide, nitrite, nitrate and fumarate, showing that this process depends on the DeltapH component of the proton motive force. These data, together with those concerning response to metals (Bencharit and Ward, 2005), support the idea that, in S. oneidensis, taxis towards electron acceptors is governed by an energy taxis mechanism. Surprisingly, energy taxis in S. oneidensis is not mediated by the PAS containing chemoreceptors but rather by a chemoreceptor (SO2240) containing a Cache domain. Four other chemoreceptors also play a minor role in this process. These results indicate that energy taxis can be mediated by new types of chemoreceptors. PMID- 19555456 TI - Transcriptional loops meet chromatin: a dual-layer network controls white-opaque switching in Candida albicans. AB - The human pathogen Candida albicans is able to undergo a reversible switch between two distinct cell types called white and opaque, which are considered different transcriptional states of cells harbouring identical genomes. The present model of switching regulation includes the bistable expression of a master switch gene that is controlled by multiple transcriptional feedback loops. Here, we show that chromatin-modifying enzymes constitute an additional important regulatory layer of morphogenetic switching. We identify eight chromatin modifiers as switching modulators. Extensive epistasis analysis maps them into at least two independent signalling pathways overlaying the known transcriptional network. Interestingly, we identify the conserved Set3/Hos2 histone deacetylase complex as a key regulator relying on the methylation status of histone H3 lysine 4 for switching modulation. Furthermore, we demonstrate that opaque to white switching is facilitated by the presence of adenine in vitro, but adenine has no effect on switching once the Set3/Hos2 complex is disrupted. Our observations postulate that chromatin modifications may serve as a means to integrate environmental or host stimuli through the underlying transcriptional circuits to determine cell fate in C. albicans. PMID- 19555458 TI - The PhyR-sigma(EcfG) signalling cascade is involved in stress response and symbiotic efficiency in Bradyrhizobium japonicum. AB - PhyR is an unusual type of response regulator consisting of a receiver domain and an extracytoplasmic function (ECF) sigma factor-like domain. It was recently described as a master regulator of general stress response in Methylobacterium extorquens. Orthologues of this regulator are present in essentially all free living Alphaproteobacteria. In most of them, phyR is genetically closely linked to a gene encoding an ECF sigma factor. Here, we investigate the role of these two regulators in the soybean symbiont Bradyrhizobium japonicum USDA110. Using deletion mutants and phenotypic assays, we showed that PhyR and the ECF sigma factor sigma(EcfG) are involved in heat shock and desiccation resistance upon carbon starvation. Both mutants had symbiotic defects on the plant hosts Glycine max (soybean) and Vigna radiata (mungbean). They induced fewer nodules than the wild type and these nodules were smaller, less pigmented, and their specific nitrogenase activity was drastically reduced 2 or 3 weeks after inoculation. Four weeks after infection, soybean nodule development caught up to a large extent whereas most mungbean nodules remained defective even 5 weeks after infection. Remarkably, both mutants triggered aberrant nodules on the different host plants with ectopically emerging roots. Microarray analysis revealed that PhyR and sigma(EcfG) control congruent regulons suggesting both regulators are part of the same signalling cascade. This finding was further substantiated by in vitro protein-protein interaction studies which are in line with a partner-switching mechanism controlling gene regulation triggered by phosphorylation of PhyR. The large number of genes of unknown function present in the PhyR/sigma(EcfG) regulon and the conspicuous symbiotic phenotype suggest that these regulators are involved in the Bradyrhizobium-legume interaction via yet undisclosed mechanisms. PMID- 19555459 TI - Heterogeneity in quorum sensing-regulated bioluminescence of Vibrio harveyi. AB - Quorum sensing (QS) refers to the ability of bacterial populations to read out the local environment for cell density and to collectively activate gene expression. Vibrio harveyi, one of the best characterized model organisms in QS, was used to address the question how single cells behave within a QS-activated community in a homogeneous environment. Analysis of the QS-regulated bioluminescence of a wild type strain revealed that even at high cell densities only 69% of the cells of the population produced bioluminescence, 25% remained dark and 6% were dead. Moreover, light intensities greatly varied from cell to cell at high population density. Addition of autoinducer to a bright liquid culture of V. harveyi increased the percentage of luminescent cells up to 98%, suggesting that V. harveyi produces and/or keeps the autoinducers at non saturating concentrations. In contrast, all living cells of a constitutive QS active mutant (DeltaluxO) produced light. We also found that QS affects biofilm formation in V. harveyi. Our data provide first evidence that a heterogeneous population produces more biofilm than a homogeneous one. It is suggested that even a QS-committed population of V. harveyi takes advantage of heterogeneity, which extends the current view of QS-regulated uniformity. PMID- 19555460 TI - The ObgE/CgtA GTPase influences the stringent response to amino acid starvation in Escherichia coli. AB - The stringent response is important for bacterial survival under stressful conditions, such as amino acid starvation, and is characterized by the accumulation of ppGpp and pppGpp. ObgE (CgtA, YhbZ) is an essential conserved GTPase in Escherichia coli and several observations have implicated the protein in the control of the stringent response. However, consequences of the protein on specific responses to amino acid starvation have not been noted. We show that ObgE binds to ppGpp with biologically relevant affinity in vitro, implicating ppGpp as an in vivo ligand of ObgE. ObgE mutants increase the ratio of pppGpp to ppGpp within the cell during the stringent response. These changes are correlated with a delayed inhibition of DNA replication by the stringent response, delayed resumption of DNA replication after release, as well as a decreased survival after amino acid deprivation. With these data, we place ObgE as an active effector of the response to amino acid starvation in vivo. Our data correlate the pppGpp/ppGpp ratio with DNA replication control under bacterial starvation conditions, suggesting a possible role for the relative balance of these two nucleotides. PMID- 19555462 TI - Review: autophagy in neurodegeneration: firefighter and/or incendiarist? AB - Autophagy is an intracellular bulk degradation system that is found ubiquitously in eukaryotes. Autophagy is responsible for the degradation of most long-lived proteins and some organelles. Cytoplasmic constituents, including organelles, are sequestered into double-membrane autophagosomes, which subsequently fuse with lysosomes where their contents are degraded. This system has been implicated in various physiological processes including protein and organelle turnover, stress response, cellular differentiation, programmed cell death and pathological conditions. Defects in the autophagy machinery might have several consequences, as they have been associated with neurodegenerative disease and different forms of cancer. Thus, autophagy occupies a crucial position within the cell's metabolism, and its modulation may represent an alternative therapeutic strategy in several pathological settings including stroke, Alzheimer's, Huntington's, Parkinson's diseases and cancer. Recently, research has begun to identify some characteristics of neuronal autophagy. The results suggest that autophagy may provide a neuroprotective mechanism. However, there is evidence showing that dysfunction of autophagy in certain pathological situations can trigger and mediate programmed cell death. Autophagy has also been defined as prime suspect cause of non-apoptotic cellular demise. However, there is now mounting evidence that autophagy and apoptosis share several common regulatory elements that are crucial in any attempt to understand the dual role of autophagy in cell death and cell survival. It will be of fundamental importance to dissect whether autophagy is primarily a strategy for survival or whether autophagy can also be a part of a cell death programme and thus contribute to cell death. Many questions are open. Is autophagy a direct death execution pathway? Is autophagy an innocent bystander? Is autophagy a defence mechanism or just a scavenger or self-clearance tool in the cell? A profound understanding of the biological effects and the mechanisms underlying autophagy in neurones might be helpful in seeking effective new treatments for neurodegenerative diseases. Here, we review the defining characteristics of autophagy with special attention to its role in neurodegenerative disorders, and recent efforts to delineate the pathway of autophagic protein degradation in neurone. PMID- 19555461 TI - The ethanolamine branch of the Kennedy pathway is essential in the bloodstream form of Trypanosoma brucei. AB - Phosphatidylethanolamine (GPEtn), a major phospholipid component of trypanosome membranes, is synthesized de novo from ethanolamine through the Kennedy pathway. Here the composition of the GPEtn molecular species in the bloodstream form of Trypanosoma brucei is determined, along with new insights into phospholipid metabolism, by in vitro and in vivo characterization of a key enzyme of the Kennedy pathway, the cytosolic ethanolamine-phosphate cytidylyltransferase (TbECT). Gene knockout indicates that TbECT is essential for growth and survival, thus highlighting the importance of the Kennedy pathway for the pathogenic stage of the African trypanosome. Phosphatiylserine decarboxylation, a potential salvage pathway, does not appear to be active in cultured bloodstream form T. brucei, and it is not upregulated even when the Kennedy pathway is disrupted. In vivo metabolic labelling and phospholipid composition analysis by ESI-MS/MS of the knockout cells confirmed a significant decrease in GPEtn species, as well as changes in the relative abundance of other phospholipid species. Reduction in GPEtn levels had a profound influence on the morphology of the mutants and it compromised mitochondrial structure and function, as well as glycosylphosphatidylinositol anchor biosynthesis. TbECT is therefore genetically validated as a potential drug target against the African trypanosome. PMID- 19555463 TI - Alpha motoneurone input changes in dystrophic MDX mice after sciatic nerve transection. AB - BACKGROUND: Duchenne muscular dystrophy (DMD) is a severe form of muscular dystrophy. At present, a lot is known about the muscular degeneration in DMD, but few studies have focused on the effects on the central nervous system. In this sense, retrograde changes in the microenvironment around motor neurones in the spinal cord may contribute to the pathogenesis of the dystrophinopathies. AIMS: The aim of this study was to investigate synaptic alterations and glial reactivity in the microenvironment close to spinal motor neurones in a DMD animal model. METHODS: Six-week-old male MDX mice were subjected to left sciatic nerve transection. The axotomy was performed after the muscular degeneration/regeneration cycles previously described in such animal models. C57BL/10 mice were used as the control. Seven days after surgery, the animals were sacrificed and the lumbar spinal cords processed for immunohistochemistry using antibodies to the major histocompatibility complex of class I (MHC I), synaptophysin, IBA-1 and glial fibrillary acidic protein (GFAP). RESULTS: MHC I expression increased in both strains after axotomy. Nevertheless, the MDX mice displayed significantly lower MHC I up-regulation. With respect to GFAP expression, the MDX mice showed greater astrogliosis as compared with C57BL/10 mice. The MDX mice displayed a significant decrease in synaptophysin expression. Indeed, the ultrastructural quantitative analysis showed more intense synaptic detachment in MDX mice, indicating a reduction in synaptic activity before and after axotomy. CONCLUSIONS: The reduction in active inputs and increased gliosis in MDX mice may be associated with the muscle degeneration/regeneration cycles that occur postnatally, and could contribute to the seriousness of the disease. PMID- 19555464 TI - Abstracts of the 2009 Meeting of the Peripheral Nerve Society. July 4-8, 2009. Wurzburg, Germany. PMID- 19555465 TI - Psychotic reactivity in borderline personality disorder. AB - OBJECTIVE: To investigate the stress relatedness and paranoia specificity of psychosis in borderline personality disorder (BPD). METHOD: Fifty-six borderline patients, 38 patients with cluster C personality disorder, 81 patients with psychotic disorder and 49 healthy controls were studied with the experience sampling method (a structured diary technique) to assess: i) appraised subjective stress and ii) intensity of psychotic experiences. RESULTS: All patient groups experienced significantly more increases in psychotic experiences in relation to daily life stress than healthy controls, borderline patients displaying the strongest reactivity. Borderline patients, moreover, reported significantly more hallucinatory reactivity than healthy controls and subjects with cluster C personality disorder. Paranoid reactivity to daily life stress did not differ between the patient groups. CONCLUSION: These results are the first to ecologically validate stress-related psychosis in BPD. However, psychotic reactivity was not limited to expression of paranoia but involved a broader range of psychotic experiences including hallucinations. PMID- 19555466 TI - Abstracts of the XXII Congress of the International Society of Thrombosis and Haemostasis. Boston, Massachusetts, USA. July 11-16, 2009. PMID- 19555467 TI - PairWise Neighbours database: overlaps and spacers among prokaryote genomes. AB - BACKGROUND: Although prokaryotes live in a variety of habitats and possess different metabolic and genomic complexity, they have several genomic architectural features in common. The overlapping genes are a common feature of the prokaryote genomes. The overlapping lengths tend to be short because as the overlaps become longer they have more risk of deleterious mutations. The spacers between genes tend to be short too because of the tendency to reduce the non coding DNA among prokaryotes. However they must be long enough to maintain essential regulatory signals such as the Shine-Dalgarno (SD) sequence, which is responsible of an efficient translation. DESCRIPTION: PairWise Neighbours is an interactive and intuitive database used for retrieving information about the spacers and overlapping genes among bacterial and archaeal genomes. It contains 1,956,294 gene pairs from 678 fully sequenced prokaryote genomes and is freely available at the URL http://genomes.urv.cat/pwneigh. This database provides information about the overlaps and their conservation across species. Furthermore, it allows the wide analysis of the intergenic regions providing useful information such as the location and strength of the SD sequence. CONCLUSION: There are experiments and bioinformatic analysis that rely on correct annotations of the initiation site. Therefore, a database that studies the overlaps and spacers among prokaryotes appears to be desirable. PairWise Neighbours database permits the reliability analysis of the overlapping structures and the study of the SD presence and location among the adjacent genes, which may help to check the annotation of the initiation sites. PMID- 19555468 TI - Ethnobotanical survey of trees in Fundong, Northwest Region, Cameroon. AB - Ethnobotanical investigations were conducted in Fundong Central Subdivision in the Northwest Region of Cameroon to identify trees growing in the area and collect information on their uses by the local people. This research covered a period of 12 months from May 2007 to April 2008. Ethnobotanical information was collected through the show-and-tell/semi-structured method and personal interviews during field trips. Three villages were investigated. A total of 82 tree species were identified belonging to 70 genera and 42 families. Among these species, 40 were widely used by the local people in traditional medicine to treat 48 human ailments. Tree species were also used for fuel wood, construction materials, wood carving and honey production. Leaves and barks were commonly used in traditional medicine while the wood, branches and the entire plants were commonly used for other purposes. In spite of the scarcity of natural forests in the study area, the local populations continue to depend on indigenous and exotic trees in their surroundings for their survival. There is therefore need for cultivation, protection and sustainable management of these valuable resources for rural livelihoods. PMID- 19555470 TI - Expression of prostasin and its inhibitors during colorectal cancer carcinogenesis. AB - BACKGROUND: Clinical trials where cancer patients were treated with protease inhibitors have suggested that the serine protease, prostasin, may act as a tumour suppressor. Prostasin is proteolytically activated by the serine protease, matriptase, which has a very high oncogenic potential. Prostasin is inhibited by protease nexin-1 (PN-1) and the two isoforms encoded by the mRNA splice variants of hepatocyte growth factor activator inhibitor-1 (HAI-1), HAI-1A, and HAI-1B. METHODS: Using quantitative RT-PCR, we have determined the mRNA levels for prostasin and PN-1 in colorectal cancer tissue (n = 116), severe dysplasia (n = 13), mild/moderate dysplasia (n = 93), and in normal tissue from the same individuals. In addition, corresponding tissues were examined from healthy volunteers (n = 23). A part of the cohort was further analysed for the mRNA levels of the two variants of HAI-1, here denoted HAI-1A and HAI-1B. mRNA levels were normalised to beta-actin. Immunohistochemical analysis of prostasin and HAI 1 was performed on normal and cancer tissue. RESULTS: The mRNA level of prostasin was slightly but significantly decreased in both mild/moderate dysplasia (p < 0.001) and severe dysplasia (p < 0.01) and in carcinomas (p < 0.05) compared to normal tissue from the same individual. The mRNA level of PN-1 was more that two fold elevated in colorectal cancer tissue as compared to healthy individuals (p < 0.001) and elevated in both mild/moderate dysplasia (p < 0.01), severe dysplasia (p < 0.05) and in colorectal cancer tissue (p < 0.001) as compared to normal tissue from the same individual. The mRNA levels of HAI-1A and HAI-1B mRNAs showed the same patterns of expression. Immunohistochemistry showed that prostasin is located mainly on the apical plasma membrane in normal colorectal tissue. A large variation was found in the degree of polarization of prostasin in colorectal cancer tissue. CONCLUSION: These results show that the mRNA level of PN-1 is significantly elevated in colorectal cancer tissue. Future studies are required to clarify whether down-regulation of prostasin activity via up regulation of PN-1 is causing the malignant progression or if it is a consequence of it. PMID- 19555469 TI - A prospective study of androgen levels, hormone-related genes and risk of rheumatoid arthritis. AB - INTRODUCTION: Rheumatoid arthritis (RA) is more common in females than males and sex steroid hormones may in part explain this difference. We conducted a case control study nested within two prospective studies to determine the associations between plasma steroid hormones measured prior to RA onset and polymorphisms in the androgen receptor (AR), estrogen receptor 2 (ESR2), aromatase (CYP19) and progesterone receptor (PGR) genes and RA risk. METHODS: We genotyped AR, ESR2, CYP19, PGR SNPs and the AR CAG repeat in RA case-control studies nested within the Nurses' Health Study (NHS), NHS II (449 RA cases, 449 controls) and the Women's Health Study (72 cases, and 202 controls). All controls were matched on cohort, age, Caucasian race, menopausal status, and postmenopausal hormone use. We measured plasma dehydroepiandrosterone sulfate (DHEAS), testosterone, and sex hormone binding globulin in 132 pre-RA samples and 396 matched controls in the NHS cohorts. We used conditional logistic regression models adjusted for potential confounders to assess RA risk. RESULTS: Mean age of RA diagnosis was 55 years in both cohorts; 58% of cases were rheumatoid factor positive at diagnosis. There was no significant association between plasma DHEAS, total testosterone, or calculated free testosterone and risk of future RA. There was no association between individual variants or haplotypes in any of the genes and RA or seropositive RA, nor any association for the AR CAG repeat. CONCLUSIONS: Steroid hormone levels measured at a single time point prior to RA onset were not associated with RA risk in this study. Our findings do not suggest that androgens or the AR, ESR2, PGR, and CYP19 genes are important to RA risk in women. PMID- 19555471 TI - Brain classification reveals the right cerebellum as the best biomarker of dyslexia. AB - BACKGROUND: Developmental dyslexia is a specific cognitive disorder in reading acquisition that has genetic and neurological origins. Despite histological evidence for brain differences in dyslexia, we recently demonstrated that in large cohort of subjects, no differences between control and dyslexic readers can be found at the macroscopic level (MRI voxel), because of large variances in brain local volumes. In the present study, we aimed at finding brain areas that most discriminate dyslexic from control normal readers despite the large variance across subjects. After segmenting brain grey matter, normalizing brain size and shape and modulating the voxels' content, normal readers' brains were used to build a 'typical' brain via bootstrapped confidence intervals. Each dyslexic reader's brain was then classified independently at each voxel as being within or outside the normal range. We used this simple strategy to build a brain map showing regional percentages of differences between groups. The significance of this map was then assessed using a randomization technique. RESULTS: The right cerebellar declive and the right lentiform nucleus were the two areas that significantly differed the most between groups with 100% of the dyslexic subjects (N = 38) falling outside of the control group (N = 39) 95% confidence interval boundaries. The clinical relevance of this result was assessed by inquiring cognitive brain-based differences among dyslexic brain subgroups in comparison to normal readers' performances. The strongest difference between dyslexic subgroups was observed between subjects with lower cerebellar declive (LCD) grey matter volumes than controls and subjects with higher cerebellar declive (HCD) grey matter volumes than controls. Dyslexic subjects with LCD volumes performed worse than subjects with HCD volumes in phonologically and lexicon related tasks. Furthermore, cerebellar and lentiform grey matter volumes interacted in dyslexic subjects, so that lower and higher lentiform grey matter volumes compared to controls differently modulated the phonological and lexical performances. Best performances (observed in controls) corresponded to an optimal value of grey matter and they dropped for higher or lower volumes. CONCLUSION: These results provide evidence for the existence of various subtypes of dyslexia characterized by different brain phenotypes. In addition, behavioural analyses suggest that these brain phenotypes relate to different deficits of automatization of language based processes such as grapheme/phoneme correspondence and/or rapid access to lexicon entries. PMID- 19555472 TI - Breast cancer stem cells: tools and models to rely on. AB - There is increasing evidence for the "cancer stem cell (CSC) hypothesis", which holds that cancers are driven by a cellular component that has stem cell properties, including self-renewal, tumorigenicity and multi-lineage differentiation capacity. Researchers and oncologists see in this model an explanation as to why cancer may be so difficult to cure, as well as a promising ground for novel therapeutic strategies. Given the specific stem cell features of self-renewal and differentiation, which drive tumorigenesis and contribute to cellular heterogeneity, each marker and assay designed to isolate and characterize CSCs has to be functionally validated. In this review, we survey tools and markers available or promising to identify breast CSCs. We review the main models used to study breast CSCs and how they challenge the CSC hypothesis. PMID- 19555473 TI - Ethical psychiatry in an uncertain world: conversations and parallel truths. AB - Psychiatric practice is often faced with complex situations that seem to pose serious moral dilemmas for practitioners. Methods for solving these dilemmas have included the development of more objective rules to guide the practitioner such as utilitarianism and deontology. A more modern variant on this objective model has been 'Principlism' where 4 mid level rules are used to help solve these complex problems. In opposition to this, there has recently been a focus on more subjective criteria for resolving complex moral dilemmas. In particular, virtue ethics has been posited as a more sensitive method for helping doctors to reason their way through difficult ethical issues. Here the focus is on the character traits of the practitioner. Bloch and Green advocated another way whereby more objective methods such as Principlism and virtue ethics are combined to produce what they considered sound moral reasoning in psychiatrists. This paper points out some difficulties with this approach and instead suggests that a better model of ethical judgment could be developed through the use of narratives or stories. This idea puts equal prima facie value on the patient's and the psychiatrist's version of the dilemma they are faced with. It has the potential to lead to a more genuine empathy and reflective decision-making. PMID- 19555474 TI - Significant differences in the use of healthcare resources of native-born and foreign born in Spain. AB - BACKGROUND: In the last decade, the number of foreign residents in Spain has doubled and it has become one of the countries in the European Union with the highest number of immigrants There is no doubt that the health of the immigrant population has become a relevant subject from the point of view of public healthcare. Our study aimed at describing the potential inequalities in the use of healthcare resources and in the lifestyles of the resident immigrant population of Spain. METHODS: Cross-sectional, epidemiological study from the Spanish National Health Survey (NHS) in 2006, from the Ministry of Health and Consumer Affairs. We have worked with individualized secondary data, collected in the Spanish National Health Survey carried out in 2006 and 2007 (SNHS-06), from the Ministry of Health and Consumer Affairs. The format of the SNHS-06 has been adapted to the requirements of the European project for the carrying out of health surveys. RESULTS: The economic immigrant population resident in Spain, present diseases that are similar to those of the indigenous population. The immigrant population shows significantly lower values in the consumption of alcohol, tobacco and physical activity (OR = 0.76; CI 95%: 0.65-0.89, they nonetheless perceive their health condition as worse than that reported by the autochthonous population (OR = 1.63, CI 95%: 1.34-1.97). The probability of the immigrant population using emergency services in the last 12 months was significantly greater than that of the autochthonous population (OR = 1.31, CI 95%: 1.12-1.54). This situation repeats itself when analyzing hospitalization data, with values of probability of being hospitalized greater among immigrants (OR = 1.39, CI 95%: 1.07-1.81). CONCLUSION: The economic immigrants have better parameters in relation to lifestyles, but they have a poor perception of their health. Despite the fact that immigrant population shows higher percentages of emergency attendance and hospitalization than the indigenous population, with respect to the use of healthcare resources, their usage of healthcare resources such as drugs, influenza vaccinations or visits to the dentist is lower. PMID- 19555475 TI - The relationship between effectiveness and costs measured by a risk-adjusted case mix system: multicentre study of Catalonian population data bases. AB - BACKGROUND: The main objective of this study is to measure the relationship between morbidity, direct health care costs and the degree of clinical effectiveness (resolution) of health centres and health professionals by the retrospective application of Adjusted Clinical Groups in a Spanish population setting. The secondary objectives are to determine the factors determining inadequate correlations and the opinion of health professionals on these instruments. METHODS/DESIGN: We will carry out a multi-centre, retrospective study using patient records from 15 primary health care centres and population data bases. The main measurements will be: general variables (age and sex, centre, service [family medicine, paediatrics], and medical unit), dependent variables (mean number of visits, episodes and direct costs), co-morbidity (Johns Hopkins University Adjusted Clinical Groups Case-Mix System) and effectiveness.The totality of centres/patients will be considered as the standard for comparison. The efficiency index for visits, tests (laboratory, radiology, others), referrals, pharmaceutical prescriptions and total will be calculated as the ratio: observed variables/variables expected by indirect standardization.The model of cost/patient/year will differentiate fixed/semi-fixed (visits) costs of the variables for each patient attended/year (N = 350,000 inhabitants). The mean relative weights of the cost of care will be obtained. The effectiveness will be measured using a set of 50 indicators of process, efficiency and/or health results, and an adjusted synthetic index will be constructed (method: percentile 50).The correlation between the efficiency (relative-weights) and synthetic (by centre and physician) indices will be established using the coefficient of determination. The opinion/degree of acceptance of physicians (N = 1,000) will be measured using a structured questionnaire including various dimensions. STATISTICAL ANALYSIS: multiple regression analysis (procedure: enter), ANCOVA (method: Bonferroni's adjustment) and multilevel analysis will be carried out to correct models. The level of statistical significance will be p < 0.05. PMID- 19555476 TI - A gene signature for post-infectious chronic fatigue syndrome. AB - BACKGROUND: At present, there are no clinically reliable disease markers for chronic fatigue syndrome. DNA chip microarray technology provides a method for examining the differential expression of mRNA from a large number of genes. Our hypothesis was that a gene expression signature, generated by microarray assays, could help identify genes which are dysregulated in patients with post-infectious CFS and so help identify biomarkers for the condition. METHODS: Human genome-wide Affymetrix GeneChip arrays (39,000 transcripts derived from 33,000 gene sequences) were used to compare the levels of gene expression in the peripheral blood mononuclear cells of male patients with post-infectious chronic fatigue (n = 8) and male healthy control subjects (n = 7). RESULTS: Patients and healthy subjects differed significantly in the level of expression of 366 genes. Analysis of the differentially expressed genes indicated functional implications in immune modulation, oxidative stress and apoptosis. Prototype biomarkers were identified on the basis of differential levels of gene expression and possible biological significance CONCLUSION: Differential expression of key genes identified in this study offer an insight into the possible mechanism of chronic fatigue following infection. The representative biomarkers identified in this research appear promising as potential biomarkers for diagnosis and treatment. PMID- 19555477 TI - Role of human papillomavirus and its detection in potentially malignant and malignant head and neck lesions: updated review. AB - Head and neck malignancies are characterized by a multiphasic and multifactorial etiopathogenesis. Tobacco and alcohol consumption are the most common risk factors for head and neck malignancy. Other factors, including DNA viruses, especially human papilloma virus (HPV), may also play a role in the initiation or development of these lesions. The pathways of HPV transmission in the head and neck mucosal lesions include oral-genital contact, more than one sexual partner and perinatal transmission of HPV to the neonatal child. The increase in prevalence of HPV infection in these lesions may be due to wider acceptance of oral sex among teenagers and adults as this is perceived to be a form of safe sex. The prevalence of HPV in benign lesions as well as malignancies has been assessed by many techniques. Among these, the polymerase chain reaction is the most sensitive method. Review of literature reveals that HPV may be a risk factor for malignancies, but not in all cases. For confirmation of the role of HPV in head and neck squamous cell carcinoma, large population studies are necessary in an assortment of clinical settings. Prophylactic vaccination against high-risk HPV types eventually may prevent a significant number of cervical carcinomas. Of the two vaccines currently available, Gardasil (Merck & Co., Inc.) protects against HPV types 6, 11, 16 and 18, while the other vaccine, Cervarix (GlaxoSmithKline, Rixensart, Belgium) protects against HPV types 16 and 18 only. However, the HPV vaccine has, to the best of our knowledge, not been tried in head and neck carcinoma. The role of HPV in etiopathogenesis, prevalence in benign and malignant lesions of this area and vaccination strategies are briefly reviewed here. PMID- 19555478 TI - Tissue-specific and neural activity-regulated expression of human BDNF gene in BAC transgenic mice. AB - BACKGROUND: Brain-derived neurotrophic factor (BDNF) is a small secreted protein that has important roles in the developing and adult nervous system. Altered expression or changes in the regulation of the BDNF gene have been implicated in a variety of human nervous system disorders. Although regulation of the rodent BDNF gene has been extensively investigated, in vivo studies regarding the human BDNF gene are largely limited to postmortem analysis. Bacterial artificial chromosome (BAC) transgenic mice harboring the human BDNF gene and its regulatory flanking sequences constitute a useful tool for studying human BDNF gene regulation and for identification of therapeutic compounds modulating BDNF expression. RESULTS: In this study we have generated and analyzed BAC transgenic mice carrying 168 kb of the human BDNF locus modified such that BDNF coding sequence was replaced with the sequence of a fusion protein consisting of N terminal BDNF and the enhanced green fluorescent protein (EGFP). The human BDNF BAC construct containing all BDNF 5' exons preceded by different promoters recapitulated the expression of endogenous BDNF mRNA in the brain and several non neural tissues of transgenic mice. All different 5' exon-specific BDNF-EGFP alternative transcripts were expressed from the transgenic human BDNF-BAC construct, resembling the expression of endogenous BDNF. Furthermore, BDNF-EGFP mRNA was induced upon treatment with kainic acid in a promotor-specific manner, similarly to that of the endogenous mouse BDNF mRNA. CONCLUSION: Genomic region covering 67 kb of human BDNF gene, 84 kb of upstream and 17 kb of downstream sequences is sufficient to drive tissue-specific and kainic acid-induced expression of the reporter gene in transgenic mice. The pattern of expression of the transgene is highly similar to BDNF gene expression in mouse and human. This is the first study to show that human BDNF gene is regulated by neural activity. PMID- 19555480 TI - Characterisation of Campylobacter jejuni genes potentially involved in phosphonate degradation. AB - Potential biological roles of the Campylobacter jejuni genes cj0641, cj0774c and cj1663 were investigated. The proteins encoded by these genes showed sequence similarities to the phosphonate utilisation PhnH, K and L gene products of Escherichia coli. The genes cj0641, cj0774c and cj1663 were amplified from the pathogenic C. jejuni strain 81116, sequenced, and cloned into pGEM-T Easy vectors. Recombinant plasmids were used to disrupt each one of the genes by inserting a kanamycin resistance (KmR) cassette employing site-directed mutagenesis or inverse PCR. Campylobacter jejuni 81116 isogenic mutants were generated by integration of the mutated genes into the genome of the wild-type strain. The C. jejuni mutants grew on primary isolation plates, but they could not be purified by subsequent passages owing to cell death. The mutant C. jejuni strains survived and proliferated in co-cultures with wild-type bacteria or in media in which wild-type C. jejuni had been previously grown. PCR analyses of mixed wild-type/mutant cultures served to verify the presence of the mutated gene in the genome of a fraction of the total bacterial population. The data suggested that each mutation inactivated a gene essential for survival. Rates of phosphonate catabolism in lysates of E. coli strain DH5 alpha were determined using proton nuclear magnetic resonance spectroscopy. Whole-cell lysates of the wild-type degraded phosphonoacetate, phenylphosphonate and aminomethylphosphonate. Significant differences in the rates of phosphonate degradation were observed between lysates of wild-type E. coli, and of bacteria transformed with each one of the vectors carrying one of the C. jejuni genes, suggesting that these genes were involved in phosphonate catabolism. PMID- 19555479 TI - Analysis of circulating hem-endothelial marker RNA levels in preterm infants. AB - BACKGROUND: Circulating endothelial cells may serve as novel markers of angiogenesis. These include a subset of hem-endothelial progenitor cells that play a vital role in vascular growth and repair. The presence and clinical implications of circulating RNA levels as an expression for hematopoietic and endothelial-specific markers have not been previously evaluated in preterm infants. This study aims to determine circulating RNA levels of hem-endothelial marker genes in peripheral blood of preterm infants and begin to correlate these findings with prenatal complications. METHODS: Peripheral blood samples from seventeen preterm neonates were analyzed at three consecutive post-delivery time points (day 3-5, 10-15 and 30). Using quantitative reverse transcription polymerase chain reaction we studied the expression patterns of previously established hem-endothelial-specific progenitor-associated genes (AC133, Tie-2, Flk-1 (VEGFR2) and Scl/Tal1) in association with characteristics of prematurity and preterm morbidity. RESULTS: Circulating Tie-2 and SCL/Tal1 RNA levels displayed an inverse correlation to gestational age (GA). We observed significantly elevated Tie-2 levels in preterm infants born to mothers with amnionitis, and in infants with sustained brain echogenicity on brain sonography. Other markers showed similar expression patterns yet we could not demonstrate statistically significant correlations. CONCLUSION: These preliminary findings suggest that circulating RNA levels especially Tie2 and SCL decline with maturation and might relate to some preterm complication. Further prospective follow up of larger cohorts are required to establish this association. PMID- 19555481 TI - Accuracy and limitations of equations for predicting the glomerular filtration rate during follow-up of patients with non-diabetic nephropathies. PMID- 19555482 TI - Computational analysis of the interaction between transcription factors and the predicted secreted proteome of the yeast Kluyveromyces lactis. AB - BACKGROUND: Protein secretion is a cell translocation process of major biological and technological significance. The secretion and downstream processing of proteins by recombinant cells is of great commercial interest. The yeast Kluyveromyces lactis is considered a promising host for heterologous protein production. Because yeasts naturally do not secrete as many proteins as filamentous fungi, they can produce secreted recombinant proteins with few contaminants in the medium. An ideal system to address the secretion of a desired protein could be exploited among the native proteins in certain physiological conditions. By applying algorithms to the completed K. lactis genome sequence, such a system could be selected. To this end, we predicted protein subcellular locations and correlated the resulting extracellular secretome with the transcription factors that modulate the cellular response to a particular environmental stimulus. RESULTS: To explore the potential Kluyveromyces lactis extracellular secretome, four computational prediction algorithms were applied to 5076 predicted K. lactis proteins from the genome database. SignalP v3 identified 418 proteins with N-terminal signal peptides. From these 418 proteins, the Phobius algorithm predicted that 176 proteins have no transmembrane domains, and the big-PI Predictor identified 150 proteins as having no glycosylphosphatidylinositol (GPI) modification sites. WoLF PSORT predicted that the K. lactis secretome consists of 109 putative proteins, excluding subcellular targeting. The transcription regulators of the putative extracellular proteins were investigated by searching for DNA binding sites in their putative promoters. The conditions to favor expression were obtained by searching Gene Ontology terms and using graph theory. CONCLUSION: A public database of K. lactis secreted proteins and their transcription factors are presented. It consists of 109 ORFs and 23 transcription factors. A graph created from this database shows 134 nodes and 884 edges, suggesting a vast number of relationships to be validated experimentally. Most of the transcription factors are related to responses to stress such as drug, acid and heat resistance, as well as nitrogen limitation, and may be useful for inducing maximal expression of potential extracellular proteins. PMID- 19555483 TI - Percutaneous endoscopic lumbar discectomy: clinical and quality of life outcomes with a minimum 2 year follow-up. AB - BACKGROUND: Percutaneous endoscopic lumbar discectomy is a relatively new technique. Very few studies have reported the clinical outcome of percutaneous endoscopic discectomy in terms of quality of life and return to work. METHOD: 55 patients with percutaneous endoscopic lumbar discectomy done from 2002 to 2006 had their clinical outcomes reviewed in terms of the North American Spine Score (NASS), Medical Outcomes Study Short Form-36 scores (SF-36) and Pain Visual Analogue Scale (VAS) and return to work. RESULTS: The mean age was 35.6 years, the mean operative time was 55.8 minutes and the mean length of follow-up was 3.4 years. The mean hospital stay for endoscopic discectomy was 17.3 hours. There was significant reduction in the severity of back pain and lower limb symptoms (NASS and VAS, p < 0.05) at 6 months and 2 years. There was significant improvement in all aspects of the Quality of Life (SF-36, p < 0.05) scores except for general health at 6 months and 2 years postoperation. The recurrence rate was 5% (3 patients). 5% (3 patients) subsequently underwent lumbar fusion for persistent back pain. All patients returned to their previous occupation after surgery at a mean time of 24.3 days. CONCLUSION: Percutaneous endoscopic lumbar discectomy is associated with improvement in back pain and lower limb symptoms postoperation which translates to improvement in quality of life. It has the advantage that it can be performed on a day case basis with short length of hospitalization and early return to work thus improving quality of life earlier. PMID- 19555484 TI - Cough and dyspnea during bronchoconstriction: comparison of different stimuli. AB - BACKGROUND: Bronchial challenge tests are used to evaluate bronchial responsiveness in diagnosis and follow-up of asthmatic patients. Challenge induced cough has increasingly been recognized as a valuable diagnostic tool. Various stimuli and protocols have been employed. The aim of this study was to compare cough and dyspnea intensity induced by different stimuli. METHODS: Twenty asthmatic patients underwent challenge tests with methacholine, bradykinin and exercise. Cough was counted during challenge tests. Dyspnea was assessed by modified Borg scale and visual analogue scale. Statistical comparisons were performed by linear mixed-effects model. RESULTS: For cough evaluation, bradykinin was the most potent trigger (p < 0.01). In terms of dyspnea measured by Borg scale, there were no differences among stimuli (p > 0.05). By visual analogue scale, bradykinin induced more dyspnea than other stimuli (p < or = 0.04). CONCLUSION: Bradykinin seems to be the most suitable stimulus for bronchial challenge tests intended for measuring cough in association with bronchoconstriction. PMID- 19555485 TI - Health services research doctoral core competencies. AB - This manuscript presents an initial description of doctoral level core competencies for health services research (HSR). The competencies were developed by a review of the literature, text analysis of institutional accreditation self studies submitted to the Council on Education for Public Health, and a consensus conference of HSR educators from US educational institutions. The competencies are described in broad terms which reflect the unique expertise, interests, and preferred learning methods of academic HSR programs. This initial set of core competencies is published to generate further dialogue within and outside of the US about the most important learning objectives and methods for HSR training and to clarify the unique skills of HSR training program graduates. PMID- 19555486 TI - Agrobacterium rhizogenes-mediated transformation of Superroot-derived Lotus corniculatus plants: a valuable tool for functional genomics. AB - BACKGROUND: Transgenic approaches provide a powerful tool for gene function investigations in plants. However, some legumes are still recalcitrant to current transformation technologies, limiting the extent to which functional genomic studies can be performed on. Superroot of Lotus corniculatus is a continuous root cloning system allowing direct somatic embryogenesis and mass regeneration of plants. Recently, a technique to obtain transgenic L. corniculatus plants from Superroot-derived leaves through A. tumefaciens-mediated transformation was described. However, transformation efficiency was low and it took about six months from gene transfer to PCR identification. RESULTS: In the present study, we developed an A. rhizogenes-mediated transformation of Superroot-derived L. corniculatus for gene function investigation, combining the efficient A. rhizogenes-mediated transformation and the rapid regeneration system of Superroot. The transformation system using A. rhizogenes K599 harbouring pGFPGUSPlus was improved by validating some parameters which may influence the transformation frequency. Using stem sections with one node as explants, a 2-day pre-culture of explants, infection with K599 at OD(600) = 0.6, and co-cultivation on medium (pH 5.4) at 22 degrees C for 2 days enhanced the transformation frequency significantly. As proof of concept, Superroot-derived L. corniculatus was transformed with a gene from wheat encoding an Na+/H+ antiporter (TaNHX2) using the described system. Transgenic Superroot plants were obtained and had increased salt tolerance, as expected from the expression of TaNHX2. CONCLUSION: A rapid and efficient tool for gene function investigation in L. corniculatus was developed, combining the simplicity and high efficiency of the Superroot regeneration system and the availability of A. rhizogenes-mediated transformation. This system was improved by validating some parameters influencing the transformation frequency, which could reach 92% based on GUS detection. The combination of the highly efficient transformation and the regeneration system of Superroot provides a valuable tool for functional genomics studies in L. corniculatus. PMID- 19555487 TI - Difficulty in diagnosing the pathological nature of an acute fracture of the clavicle: a case report. AB - Fractures of the clavicle comprise between 5% to10% of all fractures. Medial clavicular fractures are uncommon and are normally caused by high-energy trauma. A low impact mechanism of injury should raise suspicion of a pathological fracture, but this case report highlights the difficulty in diagnosing the pathological nature of an acute fracture of the clavicle. We describe a patient who presented with a medial clavicular fracture after a simple fall but the fracture was diagnosed as pathological in retrospect four months after the initial presentation. We would also like to emphasise that the medial clavicle is the most frequent site of pathological fractures of the clavicle, and the possibility of an underlying pathological condition should be considered whenever a patient with a medial clavicular fracture is encountered. PMID- 19555488 TI - Mechanically-induced osteogenesis in the cortical bone of pre- to peripubertal stage and peri- to postpubertal stage mice. AB - BACKGROUND: Exercise during postnatal development plays a key role in determining adult bone mass and reducing the risk of fracture and osteoporosis later in life. However, the relationship between mechanically-induced osteogenesis and age is unclear. Elevated levels of estrogen during puberty may inhibit periosteal bone formation. Thus, magnitudes of mechanically-induced osteogenesis may be vary with pubertal state. METHODS: The present study uses a murine model to examine age related changes in bone formation at the femoral midshaft with voluntary exercise. Pre- to peripubertal mice aged 3 weeks and peri- to postpubertal mice aged 7 weeks were randomly divided into sedentary and exercised groups and subjected to histomorphometric comparison after 4 weeks of treatment. RESULTS: Results of the experiment indicate that exercise significantly increased osteogenesis on the periosteal and endocortical surface of the mice in the older age group (P < 0.05). Exercise had no significant effect on bone formation of mice in the younger age group, although exercised mice exhibited more bone growth on average than controls. Endocortical apposition was the primary method of bone formation for all mice in the experiment; however exercised mice in the older age group were able to add more bone on the periosteal surface than age-matched controls and exercised mice in the younger age group (P < 0.05). Medullary area increased with age, but exercised mice in both age groups had smaller medullary cavities relative to overall bone area than controls. CONCLUSION: These findings suggest that the amount and location of mechanically-induced osteogenesis differs by age during skeletal development. Late adolescence may be the optimal time to accrue bone mass and maximize bone strength. PMID- 19555489 TI - Internationally trained pharmacists in Great Britain: what do registration data tell us about their recruitment? AB - BACKGROUND: Internationally trained health professionals are an important part of the domestic workforce, but little is known about pharmacists who come to work in Great Britain. Recent changes in the registration routes onto the Register of Pharmacists of the Royal Pharmaceutical Society of Great Britain may have affected entries from overseas: reciprocal arrangements for pharmacists from Australia and New Zealand ended in June 2006; 10 new states joined the European Union in 2004 and a further two in 2007, allowing straightforward registration. AIMS: The aims of the paper are to extend our knowledge about the extent to which Great Britain is relying on the contribution of internationally trained pharmacists and to explore their routes of entry and demographic characteristics and compare them to those of pharmacists trained in Great Britain. METHODS: The August 2007 Register of Pharmacists provided the main data for analysis. Register extracts between 2002 and 2005 were also explored, allowing longitudinal comparison, and work pattern data from the 2005 Pharmacist Workforce Census were included. RESULTS: In 2007, internationally trained pharmacists represented 8.8% of the 43,262 registered pharmacists domiciled in Great Britain. The majority (40.6%) had joined the Register from Europe; 33.6% and 25.8% joined via adjudication and reciprocal arrangements. Until this entry route ended for pharmacists from Australia and New Zealand in 2006, annual numbers of reciprocal pharmacists increased. European pharmacists are younger (mean age 31.7) than reciprocal (40.0) or adjudication pharmacists (43.0), and the percentage of women among European-trained pharmacists is much higher (68%) when compared with British-trained pharmacists (56%). While only 7.1% of pharmacists registered in Great Britain have a London address, this proportion is much higher for European (13.9%), adjudication (19.5%) and reciprocal pharmacists (28.9%). The latter are more likely to work in hospitals than in community pharmacies, and all groups of internationally trained pharmacist are more likely to work full-time than British trained ones. Adjudication pharmacists appear to stay on the Register longer than their reciprocal and European colleagues. CONCLUSION: Analysis of the Register of Pharmacists provides novel insights into the origins, composition and destinations of internationally trained pharmacists. They represent a notable proportion of the Register, indicating that British employers are relying on their contribution for the delivery of pharmacy services. With the increasing mobility of health care professionals across geographical borders, it will be important to undertake primary research to gain a better understanding of the expectations, plans and experiences of pharmacists entering from outside Great Britain. PMID- 19555490 TI - Oral vaccination with a recombinant Salmonella vaccine vector provokes systemic HIV-1 subtype C Gag-specific CD4+ Th1 and Th2 cell immune responses in mice. AB - BACKGROUND: Recombinant Salmonella vaccine vectors may potentially be used to induce specific CD4+ T cell responses against foreign viral antigens. Such immune responses are required features of vaccines against pathogens such as human immunodeficiency virus type 1 (HIV-1). The aim of this study was to investigate the induction of systemic HIV-1-specific CD4+ T helper (Th) responses in mice after oral immunization with a live attenuated Salmonella vaccine vector that expressed HIV-1 subtype C Gag. Groups of BALB/c mice were vaccinated orally three times (4 weeks apart) with this recombinant Salmonella. At sacrifice, 28 days after the last immunization, systemic CD4+ Th1 and Th2 cytokine responses were evaluated by enzyme-linked immunospot assay and cytometric bead array. HIV-1 Gag specific IgG1 and IgG2a humoral responses in the serum were determined by enzyme linked immunosorbent assay. RESULTS: Mice vaccinated with the recombinant Salmonella elicited both HIV-1-specific Th1 (interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha)) and Th2 (interleukin-4 (IL-4) and interleukin-5 (IL-5)) cytokine responses. The vaccine induced 70 (IFN-gamma) spot forming units (SFUs)/10e(6) splenocytes and 238 IL-4 SFUs/10e(6) splenocytes. Splenocytes from vaccinated mice also produced high levels of Th1 and Th2 cytokines upon stimulation with a Gag CD4 peptide. The levels of IFN-gamma, TNF alpha, IL-4 and IL-5 were 7.5-, 29.1-, 26.2- and 89.3-fold above the background, respectively. Both HIV-1 Gag-specific IgG1 and IgG2a antibodies were detected in the sera of vaccinated mice. CONCLUSION: The study highlights the potential of orally-delivered attenuated Salmonella as mucosal vaccine vectors for HIV-1 Subtype C Gag to induce Gag-specific CD4+ Th1 and Th2 cellular immune responses and antibodies which may be important characteristics required for protection against HIV-1 infection. PMID- 19555492 TI - National support to public health research: a survey of European ministries. AB - BACKGROUND: Within SPHERE (Strengthening Public Health Research in Europe), a collaborative study funded by the European Commission, we have assessed the support for public health research at ministry level in European countries. METHODS: We surveyed the health and science ministries in 25 EU countries and 3 EEA countries, using a broad definition of public-health research at population level. We made over 600 phone calls and emails to identify respondents and to gain answers. We gained formal replies from 42 out of 56 ministries (73% response) in 25 countries. There were 22 completed questionnaires (from 25 ministries), 6 short answers and 11 contacts declaring that their ministries were not responsible for public health research, while in 14 ministries (both ministries in three countries) no suitable ministry contact could be found. RESULTS: In most European countries, ministries of health, or their devolved agencies, were regarded as the leading organizations. Most ministries were able to specify thematic areas for public-health research (from three to thirty), and others ministries referred to policy documents, health plans or public-health plans to define research priorities. Ministries and their agencies led on decisions for financial support of public-health research, with less involvement of other external organisations compared with the process of identifying priorities. However, the actual funds available for public health were not easily identifiable. Most ministries relied on general academic means for dissemination of results of public-health research, while ministries get information on the use of public-health research usually through informal means. Ministries made suggestions for strengthening public-health research through initiatives of their own countries and of the European Union: as well as more resources, improving coordination was most frequently suggested. CONCLUSION: There is no common approach to support for public-health research across Europe, and significant gaps in organisation and funding. Health ministries and national agencies value exchange between researchers and policy-makers, civil society organizations, and academic and public authorities, and the application of public-health research results. There would be benefits from better processes of priority setting and improved coordination for research, at regional, national and European levels. PMID- 19555491 TI - Conserved cryptic recombination signals in Vkappa gene segments are cleaved in small pre-B cells. AB - BACKGROUND: The cleavage of recombination signals (RS) at the boundaries of immunoglobulin V, D, and J gene segments initiates the somatic generation of the antigen receptor genes expressed by B lymphocytes. RS contain a conserved heptamer and nonamer motif separated by non-conserved spacers of 12 or 23 nucleotides. Under physiologic conditions, V(D)J recombination follows the "12/23 rule" to assemble functional antigen-receptor genes, i.e., cleavage and recombination occur only between RS with dissimilar spacer types. Functional, cryptic RS (cRS) have been identified in VH gene segments; these VH cRS were hypothesized to facilitate self-tolerance by mediating VH --> VHDJH replacements. At the Igkappa locus, however, secondary, de novo rearrangements can delete autoreactive VkappaJkappa joins. Thus, under the hypothesis that V-embedded cRS are conserved to facilitate self-tolerance by mediating V-replacement rearrangements, there would be little selection for Vkappa cRS. Recent studies have demonstrated that VH cRS cleavage is only modestly more efficient than V(D)J recombination in violation of the 12/23 rule and first occurs in pro-B cells unable to interact with exogenous antigens. These results are inconsistent with a model of cRS cleavage during autoreactivity-induced VH gene replacement. RESULTS: To test the hypothesis that cRS are absent from Vkappa gene segments, a corollary of the hypothesis that the need for tolerizing VH replacements is responsible for the selection pressure to maintain VH cRS, we searched for cRS in mouse Vkappa gene segments using a statistical model of RS. Scans of 135 mouse Vkappa gene segments revealed highly conserved cRS that were shown to be cleaved in the 103/BCL2 cell line and mouse bone marrow B cells. Analogous to results for VH cRS, we find that Vkappa cRS are conserved at multiple locations in Vkappa gene segments and are cleaved in pre-B cells. CONCLUSION: Our results, together with those for VH cRS, support a model of cRS cleavage in which cleavage is independent of BCR-specificity. Our results are inconsistent with the hypothesis that cRS are conserved solely to support receptor editing. The extent to which these sequences are conserved, and their pattern of conservation, suggest that they may serve an as yet unidentified purpose. PMID- 19555493 TI - Evolving e-taxonomy. PMID- 19555494 TI - Cognitive health among older adults in the United States and in England. AB - BACKGROUND: Cognitive function is a key determinant of independence and quality of life among older adults. Compared to adults in England, US adults have a greater prevalence of cardiovascular risk factors and disease that may lead to poorer cognitive function. We compared cognitive performance of older adults in the US and England, and sought to identify sociodemographic and medical factors associated with differences in cognitive function between the two countries. METHODS: Data were from the 2002 waves of the US Health and Retirement Study (HRS) (n = 8,299) and the English Longitudinal Study of Ageing (ELSA) (n = 5,276), nationally representative population-based studies designed to facilitate direct comparisons of health, wealth, and well-being. There were differences in the administration of the HRS and ELSA surveys, including use of both telephone and in-person administration of the HRS compared to only in-person administration of the ELSA, and a significantly higher response rate for the HRS (87% for the HRS vs. 67% for the ELSA). In each country, we assessed cognitive performance in non-hispanic whites aged 65 and over using the same tests of memory and orientation (0 to 24 point scale). RESULTS: US adults scored significantly better than English adults on the 24-point cognitive scale (unadjusted mean: 12.8 vs. 11.4, P < .001; age- and sex-adjusted: 13.2 vs. 11.7, P < .001). The US cognitive advantage was apparent even though US adults had a significantly higher prevalence of cardiovascular risk factors and disease. In a series of OLS regression analyses that controlled for a range of sociodemographic and medical factors, higher levels of education and wealth, and lower levels of depressive symptoms, accounted for some of the US cognitive advantage. US adults were also more likely to be taking medications for hypertension, and hypertension treatment was associated with significantly better cognitive function in the US, but not in England (P = .014 for treatment x country interaction). CONCLUSION: Despite methodological differences in the administration of the surveys in the two countries, US adults aged >/= 65 appeared to be cognitively healthier than English adults, even though they had a higher burden of cardiovascular risk factors and disease. Given the growing number of older adults worldwide, future cross-national studies aimed at identifying the medical and social factors that might prevent or delay cognitive decline in older adults would make important and valuable contributions to public health. PMID- 19555495 TI - In silico analysis of the cyclophilin repertoire of apicomplexan parasites. AB - BACKGROUND: Cyclophilins (Cyps) are peptidyl cis/trans isomerases implicated in diverse processes such as protein folding, signal transduction, and RNA processing. They are also candidate drug targets, in particular for the immunosuppressant cyclosporine A. In addition, cyclosporine is known to exhibit anti-parasitic effects on a wide range of organisms including several apicomplexa. In order to obtain new non-immunosuppressive drugs targeting apicomplexan cyclophilins, a profound knowledge of the cyclophilin repertoire of this phylum would be necessary. RESULTS: BLAST and maximum likelihood analyses identified 16 different cyclophilin subfamilies within the genomes of Cryptosporidium hominis, Toxoplasma gondii, Plasmodium falciparum, Theileria annulata, Theileria parva, and Babesia bovis. In addition to good statistical support from the phylogenetic analysis, these subfamilies are also confirmed by comparison of cyclophilin domain architecture. Within an individual genome, the number of different Cyp genes that could be deduced varies between 7-9 for Cryptosporidia and 14 for T. gondii. Many of the putative apicomplexan cyclophilins are predicted to be nuclear proteins, most of them presumably involved in RNA processing. CONCLUSION: The genomes of apicomplexa harbor a cyclophilin repertoire that is at least as complex as that of most fungi. The identification of Cyp subfamilies that are specific for lower eukaryotes, apicomplexa, or even the genus Plasmodium is of particular interest since these subfamilies are not present in host cells and might therefore represent attractive drug targets. PMID- 19555496 TI - A no-go related prefrontal negativity larger to irrelevant stimuli that are difficult to suppress. AB - BACKGROUND: There is a wide debate in the literature about whether N2/P3 effects in no-go trials reflect the inhibition of an intended action, or the absence of a negative movement-related potential typical of go trials. The aim of this study was to provide an objective measure of the suppression of irrelevant information (in a conjoined selective visual attention task) under conditions that were perfectly comparable from the viewpoint of the motoric processes involved. METHODS: Twenty-nine right-handed students took part in the study. Their EEGs were recorded from 128 scalp sites while they viewed gratings of four different spatial frequencies (from 0.75 to 6 c/deg) randomly flashed in the four upper and lower quadrants of the visual field. The tasks consisted of attending and responding to a conjunction of spatial frequency and space location. Intermediate frequencies (1.5 and 3 c/deg) acted as distracters or lures. Analysis of the ERPs elicited by the same physical stimulus, close in spatial frequency to the actual target and falling within the attended quadrant (pseudo-target) vs. a non-target location, allowed us to identify the time course and neural bases of brain activation during the suppression of irrelevant information. RESULTS: FAs were on average 9% for pseudo-targets and 0.2% for other types of lures, indicating that the former were more difficult to suppress. Target-related ERP components (occipito/temporal selection negativity, posterior P3b and precentral motor N2) were greater to pseudo-targets than other distracters. A large prefrontal negativity (370-430 ms) was also identified, much larger to pseudo-targets than non-targets (and absent in response to real targets), thus reflecting response inhibition and top-down cognitive control processes. CONCLUSION: A LORETA inverse solution identified the neural generators of this effect in the left dorsolateral prefrontal cortex (DLPF), left and right fusiform gyri and bilateral superior temporal cortices. The tentative hypothesis is advanced that these activations might reflect the modulatory effects exerted by the fronto/temporal circuit for the suppression of irrelevant information. PMID- 19555497 TI - Co-activation of hedgehog and AKT pathways promote tumorigenesis in zebrafish. AB - The zebrafish has become an important model for cancer research. Several cancer models have been established by transgenic expression of human or mouse oncogenes in zebrafish. Since it is amenable to efficient transgenesis, zebrafish have immense potential to be used for studying interaction of oncogenes and pathways at the organismal level. Using the Gal4VP16-UAS binary transgenic expression approach, we established stable transgenic lines expressing an EGFP fusion protein of an activated zebrafish Smoothened (Smoa1-EGFP). Expression of the zebrafish Smoa1-EGFP itself did not lead to tumor formation either in founder fish or subsequent generations, however, co-expressing a constitutively active human AKT1 resulted in several tumor types, including spindle cell sarcoma, rhabdomyoma, ocular melanoma, astrocytoma, and myxoma. All tumor types showed GFP expression and increased Patched 1 levels, suggesting involvement of zebrafish Smoa1 in tumorigenesis. Immunofluorescence studies showed that tumors also expressed elevated levels of phosphorylated AKT, indicating activation of the PI3K-AKT pathway. These results suggest that co-activation of the hedgehog and AKT pathways promote tumorigenesis, and that the binary transgenic approach is a useful tool for studying interaction of oncogenes and oncogenic pathways in zebrafish. PMID- 19555498 TI - Theoretical study of the Usutu virus helicase 3D structure, by means of computer aided homology modelling. AB - BACKGROUND: Usutu virus belongs to the Flaviviridae viral family and constitutes an important pathogen. The viral helicase is an ideal target for inhibitor design, since this enzyme is essential for the survival, proliferation and transmission of the virus. RESULTS: Towards a drug-design approach, the 3D model of the Usutu virus helicase structure has been designed, using conventional homology modelling techniques and the known 3D-structure of the Murray Valley Encephalitis virus helicase, of the same viral family, as template. The model was then subjected to extended molecular dynamics simulations in a periodic box, filled with explicit water molecules for 10 nanoseconds. The reliability of the model was confirmed by obtaining acceptable scores from a variety of in silico scoring tools, including Procheck and Verify3D. CONCLUSION: [corrected] The 3D model of the Usutu virus helicase exhibits in silico all known structural characteristics of the Flaviviridae viral family helicase enzymes and could provide the platform for further de novo structure-based design of novel anti Usutu agents. PMID- 19555499 TI - Evidence for a mitochondrial localization of the retinoblastoma protein. AB - BACKGROUND: The retinoblastoma protein (Rb) plays a central role in the regulation of cell cycle, differentiation and apoptosis. In cancer cells, ablation of Rb function or its pathway is a consequence of genetic inactivation, viral oncoprotein binding or deregulated hyperphosphorylation. Some recent data suggest that Rb relocation could also account for the regulation of its tumor suppressor activity, as is the case for other tumor suppressor proteins, such as p53. RESULTS: In this reported study, we present evidence that a fraction of the total amount of Rb protein can localize to the mitochondria in proliferative cells taken from both rodent and human cells. This result is also supported by the use of Rb siRNAs, which substantially reduced the amount of mitochondrial Rb, and by acellular assays, in which [35S]-Methionine-labeled Rb proteins bind strongly to mitochondria isolated from rat liver. Moreover, endogenous Rb is found in an internal compartment of the mitochondria, within the inner-membrane. This is consistent with the protection of Rb from alkaline treatment, which destroys any interaction of proteins that are weakly bound to mitochondria. CONCLUSION: Although a few data regarding an unspecific cytosolic localization of Rb protein have been reported for some tumor cells, our results are the first evidence of a mitochondrial localization of Rb. The mitochondrial localization of Rb is observed in parallel with its classic nuclear location and paves the way for the study of potential as-yet-unknown roles of Rb at this site. PMID- 19555500 TI - Analysis of salivary transcripts and antigens of the sand fly Phlebotomus arabicus. AB - BACKGROUND: Sand fly saliva plays an important role in blood feeding and Leishmania transmission as it was shown to increase parasite virulence. On the other hand, immunity to salivary components impedes the establishment of infection. Therefore, it is most desirable to gain a deeper insight into the composition of saliva in sand fly species which serve as vectors of various forms of leishmaniases. In the present work, we focused on Phlebotomus (Adlerius) arabicus, which was recently shown to transmit Leishmania tropica, the causative agent of cutaneous leishmaniasis in Israel. RESULTS: A cDNA library from salivary glands of P. arabicus females was constructed and transcripts were sequenced and analyzed. The most abundant protein families identified were SP15-like proteins, ParSP25-like proteins, D7-related proteins, yellow-related proteins, PpSP32-like proteins, antigen 5-related proteins, and 34 kDa-like proteins. Sequences coding for apyrases, hyaluronidase and other putative secreted enzymes were also represented, including endonuclease, phospholipase, pyrophosphatase, amylase and trehalase. Mass spectrometry analysis confirmed the presence of 20 proteins predicted to be secreted in the salivary proteome. Humoral response of mice bitten by P. arabicus to salivary antigens was assessed and many salivary proteins were determined to be antigenic. CONCLUSION: This transcriptomic analysis of P. arabicus salivary glands is the first description of salivary proteins of a sand fly in the subgenus Adlerius. Proteomic analysis of P. arabicus salivary glands produced the most comprehensive account in a single sand fly species to date. Detailed information and phylogenetic relationships of the salivary proteins are provided, expanding the knowledge base of molecules that are likely important factors of sand fly-host and sand fly-Leishmania interactions. Enzymatic and immunological investigations further demonstrate the value of functional transcriptomics in advancing biological and epidemiological research that can impact leishmaniasis. PMID- 19555501 TI - Association of selected SNP with carcass and taste panel assessed meat quality traits in a commercial population of Aberdeen Angus-sired beef cattle. AB - BACKGROUND: The purpose of this study was to evaluate the effects of eight single nucleotide polymorphisms (SNP), previously associated with meat and milk quality traits in cattle, in a population of 443 commercial Aberdeen Angus-cross beef cattle. The eight SNP, which were located within five genes: mu-calpain (CAPN1), calpastatin (CAST), leptin (LEP), growth hormone receptor (GHR) and acylCoA:diacylglycerol acyltransferase 1 (DGAT1), are included in various commercial tests for tenderness, fatness, carcass composition and milk yield/quality. METHODS: A total of 27 traits were examined, 19 relating to carcass quality, such as carcass weight and fatness, one mechanical measure of tenderness, and the remaining seven were sensory traits, such as flavour and tenderness, assessed by a taste panel. RESULTS: An SNP in the CAPN1 gene, CAPN316, was significantly associated with tenderness measured by both the tenderometer and the taste panel as well as the weight of the hindquarter, where animals inheriting the CC genotype had more tender meat and heavier hindquarters. An SNP in the leptin gene, UASMS2, significantly affected overall liking, where animals with the TT genotype were assigned higher scores by the panellists. The SNP in the GHR gene was significantly associated with odour, where animals inheriting the AA genotype produced steaks with an intense odour when compared with the other genotypes. Finally, the SNP in the DGAT1 gene was associated with sirloin weight after maturation and fat depth surrounding the sirloin, with animals inheriting the AA genotype having heavier sirloins and more fat. CONCLUSION: The results of this study confirm some previously documented associations. Furthermore, novel associations have been identified which, following validation in other populations, could be incorporated into breeding programmes to improve meat quality. PMID- 19555503 TI - Cytochrome-P450 enzymes and autoimmunity: expansion of the relationship and introduction of free radicals as the link. AB - The Cytochrome-P-450 enzymes (CYP) are among the most important xenobiotic metabolizing enzymes, which produce reactive oxygen species (ROS) as the result of metabolizing xenobiotics. ROS are believed to play important roles in the pathophysiology of autoimmune diseases. ROS can alter the structure of cellular antigens to produce a "neo-antigen" which could mount an autoimmune response against the original antigen through molecular mimicry. ROS are involved in apoptosis, activation of antigen presenting cells and initiation or amplification of diverse immunologic reactions. Taking all these facts together, it could be speculated that CYP may be involved in the initiation and/or amplification of autoimmune phenomena. PMID- 19555502 TI - Exercise therapy for the management of osteoarthritis of the hip joint: a systematic review. AB - INTRODUCTION: Recent guidelines pertaining to exercise for individuals with osteoarthritis have been released. These guidelines have been based primarily on studies of knee-joint osteoarthritis. The current study was focused on the hip joint, which has different biomechanical features and risk factors for osteoarthritis and has received much less attention in the literature. The purpose was to conduct a systematic review of the literature to evaluate the exercise programs used in intervention studies focused solely on hip-joint osteoarthritis, to decide whether their exercise regimens met the new guidelines, and to determine the level of support for exercise-therapy interventions in the management of hip-joint osteoarthritis. METHODS: A systematic literature search of 14 electronic databases was undertaken to identify interventions that used exercise therapy as a treatment modality for hip osteoarthritis. The quality of each article was critically appraised and graded according to standardized methodologic approaches. A 'pattern-of-evidence' approach was used to determine the overall level of evidence in support of exercise-therapy interventions for treating hip osteoarthritis. RESULTS: More than 4,000 articles were identified, of which 338 were considered suitable for abstract review. Of these, only 6 intervention studies met the inclusion criteria. Few well-designed studies specifically investigated the use of exercise-therapy management on hip-joint osteoarthritis. Insufficient evidence was found to suggest that exercise therapy can be an effective short-term management approach for reducing pain levels, improving joint function and the quality of life. CONCLUSIONS: Limited information was available on which conclusions regarding the efficacy of exercise could be clearly based. No studies met the level of exercise recommended for individuals with osteoarthritis. High-quality trials are needed, and further consideration should be given to establishing the optimal exercises and exposure levels necessary for achieving long-term gains in the management of osteoarthritis of the hip. PMID- 19555504 TI - The change in attitude and knowledge of health care personnel and general population following trainings provided during integration of mental health in Primary Health Care in Iran: a systematic review. AB - BACKGROUND: Mental health has been integrated in the primary health care program in small cities and villages of Iran in a national level since the late 1980s. We performed a systematic review of literature to investigate the effect of education on change in attitude and knowledge of mental health care providers and the population covered in the program during the recent two decades in Iran. METHODS: Electronic bibliographic databases including Pubmed, PsycINFO and EMBase as well as the main Iranian databases (Scientific Information Database, IranMedex, IranPsych, and IranDoc) were searched. Additionally, hand searching, personal contacts and tracking of reference lists were performed. All of the studies which compared the attitude and knowledge of the related population before and after an educational intervention were recruited. RESULTS: Six articles met the inclusion criteria and entered the review. All of these studies showed an improvement in the attitude and knowledge of the studied population. Although the studies were different in many respects, a meta-analysis on the two more similar studies showed a significant effect of training on long term improvement of the knowledge and attitude of the population. CONCLUSION: A short term training improved knowledge and attitude of the population and health personnel immediately after the intervention. There is also evidence for a long term change in the attitude and knowledge of general population after short term training. PMID- 19555505 TI - Antimalarial activity of a cis-terpenone. AB - BACKGROUND: Malaria is the third most prevalent cause of infectious disease in the world. Resistance of the parasite to classical drugs makes the discovery of new and effective drugs more urgent. The oxidized derivative of hydroxy-cis terpenone (OHCT) is a synthetic molecule that is not toxic to cultured human liver cells at concentrations as high as 60 microM and inhibits activity of cytochrome P450s that metabolize many drugs. METHODS: OHCT activity against chloroquine-sensitive and -resistant strains of Plasmodium falciparum, and a P. falciparum clone that is partially resistant to artemisinin was assayed in vitro. RESULTS: OHCT at nanomolar concentrations was effective against all intraerythrocytic stages of P. falciparum and exhibited activity in vitro against both chloroquine-sensitive and -resistant strains of P. falciparum as well as a P. falciparum clone that is partially resistant to artemisinin. Moreover, OHCT exhibited potent activity against gametocytes, the form that is transmitted by mosquitoes and essential for the spread of malaria. CONCLUSION: OHCT displays strong growth inhibitory activity against all stages of P. falciparum and no evidence of toxicity to human cells in culture. It is easily synthesized and has the potential for inhibiting metabolism of drugs used in combination therapies. PMID- 19555506 TI - Knowledge, attitudes and practice of breast cancer screening among female health workers in a Nigerian urban city. AB - BACKGROUND: Late presentation has been observed as the hallmark of breast cancer in Nigerian women and an earlier onset has been reported in this population. This study was designed to assess the awareness of female health workers about risk factors and screening methods for early detection of breast cancer. METHODS: A cross-sectional descriptive study was carried out among female health workers in the two major government health institutions in Benin City, Edo State capital in Nigeria.Data analysis was by SPSS version 10 and test of significance was done with differences considered significant at p < 0.05. RESULTS: Three hundred and ninety-three (393) female health workers out of five hundred and five eligible subjects completed and returned the questionnaires, giving a response rate of 77.8%. One hundred and two (26%) were Doctors, two hundred and fifty-four (64.6%) Nurses, and thirty-seven (9.4%) were Radiographers, Laboratory Scientists and Pharmacists. A high proportion of our respondents had very poor knowledge about risk factors for breast cancer (55%). The awareness of mammography as a diagnostic method was very high (80.7%), but an extremely low knowledge of mammography as a screening method was found. Mammography practice of only 3.1% was found among those above 40 years of age who qualify for routine annual screening. Relatively low knowledge (45.5%) about Breast Self Examination (BSE) as a screening method was found. CONCLUSION: These female health workers who are expected to act as role models and educate the public had poor knowledge of risk factors for breast cancer and practice of breast cancer screening. There is very urgent need for regular update courses for health workers concerning breast cancer education including screening methods. PMID- 19555507 TI - Distribution and seasonality of rhinovirus and other respiratory viruses in a cross-section of asthmatic children in Trinidad, West Indies. AB - BACKGROUND: Childhood asthma in the Caribbean is advancing in prevalence and morbidity. Though viral respiratory tract infections are reported triggers for exacerbations, information on these infections with asthma is sparse in Caribbean territories. We examined the distribution of respiratory viruses and their association with seasons in acute and stable asthmatic children in Trinidad. METHODS: In a cross-sectional study of 70 wheezing children attending the emergency department for nebulisation and 80 stable control subjects (2 to 16 yr of age) in the asthma clinic, nasal specimens were collected during the dry (n = 38, January to May) and rainy (n = 112, June to December) seasons. A multitarget, sensitive, specific high-throughput Respiratory MultiCode assay tested for respiratory-virus sequences for eight distinct groups: human rhinovirus, respiratory syncytial virus, parainfluenza virus, influenza virus, metapneumovirus, adenovirus, coronavirus, and enterovirus. RESULTS: Wheezing children had a higher [chi(2 )= 5.561, p = 0.018] prevalence of respiratory viruses compared with stabilized asthmatics (34.3% (24) versus (vs.) 17.5% (14)). Acute asthmatics were thrice as likely to be infected with a respiratory virus (OR = 2.5, 95% CI = 1.2 - 5.3). The predominant pathogens detected in acute versus stable asthmatics were the rhinovirus (RV) (n = 18, 25.7% vs. n = 7, 8.8%; p = 0.005), respiratory syncytial virus B (RSV B) (n = 2, 2.9% vs. n = 4, 5.0%), and enterovirus (n = 1, 1.4% vs. n = 2, 2.5%). Strong odds for rhinoviral infection were observed among nebulised children compared with stable asthmatics (p = 0.005, OR = 3.6, 95% CI = 1.4 - 9.3,). RV was prevalent throughout the year (Dry, n = 6, 15.8%; Rainy, n = 19, 17.0%) and without seasonal association [chi(2 )= 0.028, p = 0.867]. However it was the most frequently detected virus [Dry = 6/10, (60.0%); Rainy = 19/28, (67.9%)] in both seasons. CONCLUSION: Emergent wheezing illnesses during childhood can be linked to infection with rhinovirus in Trinidad's tropical environment. Viral-induced exacerbations of asthma are independent of seasons in this tropical climate. Further clinical and virology investigations are recommended on the role of infections with the rhinovirus in Caribbean childhood wheeze. PMID- 19555508 TI - Selection and evaluation of reference genes for improved interrogation of microbial transcriptomes: case study with the extremophile Acidithiobacillus ferrooxidans. AB - BACKGROUND: Normalization is a prerequisite for accurate real time PCR (qPCR) expression analysis and for the validation of microarray profiling data in microbial systems. The choice and use of reference genes that are stably expressed across samples, experimental conditions and designs is a key consideration for the accurate interpretation of gene expression data. RESULTS: Here, we evaluate a carefully selected set of reference genes derived from previous microarray-based transcriptional profiling experiments performed on Acidithiobacillus ferrooxidans and identify a set of genes with minimal variability under five different experimental conditions that are frequently used in Acidithiobacilli research. Suitability of these and other previously reported reference genes to monitor the expression of four selected target genes from A. ferrooxidans grown with different energy sources was investigated. Utilization of reference genes map, rpoC, alaS and era results in improved interpretation of gene expression profiles in A. ferrooxidans. CONCLUSION: This investigation provides a validated set of reference genes for studying A. ferrooxidans gene expression under typical biological conditions and an initial point of departure for exploring new experimental setups in this microorganism and eventually in other closely related Acidithiobacilli. The information could also be of value for future transcriptomic experiments in other bacterial systems. PMID- 19555509 TI - Leisure-time physical activity, cardiorespiratory fitness and feelings of hopelessness in men. AB - BACKGROUND: Leisure-time physical activity (LTPA) and cardiorespiratory fitness contribute to mental health. Hopelessness has been linked to impaired mental health, cardiovascular events and mortality. Previous studies have focused on physical exercise and depression. We examined the associations of LTPA and cardiorespiratory fitness with feelings of hopelessness. METHODS: In this cross sectional study leisure-time physical activity, maximal oxygen uptake (VO2max), hopelessness and cardiovascular risk factors were assessed in a population-based cohort of 2428 men aged 42 - 60 years old at baseline. RESULTS: Men feeling more hopeless about their future and reaching goals were less physically active, less fit and had a higher prevalence of many cardiovascular risk factors than men with lower levels of hopelessness. In a logistic regression model adjusted for age, smoking, alcohol consumption, cardiovascular disease and socioeconomic status, men engaging in less than 60 min/week of moderate-to-vigorous LTPA were 37% (95% CI 11 - 67%) more likely to feel hopeless than those engaging in at least 2.5 h/wk of LTPA. After further adjusting for elevated depressive symptoms the association of LTPA and hopelessness remained significant. VO2max was also associated with hopelessness, but not after adjustment for depressive symptoms. CONCLUSION: Moderate and vigorous LTPA and cardiorespiratory fitness were inversely associated with hopelessness in these middle-aged men. These findings suggest that physical inactivity and poor cardiorespiratory fitness is an important associate of hopelessness, a distinct element of low subjective well being. PMID- 19555510 TI - iBsu1103: a new genome-scale metabolic model of Bacillus subtilis based on SEED annotations. AB - BACKGROUND: Bacillus subtilis is an organism of interest because of its extensive industrial applications, its similarity to pathogenic organisms, and its role as the model organism for Gram-positive, sporulating bacteria. In this work, we introduce a new genome-scale metabolic model of B. subtilis 168 called iBsu1103. This new model is based on the annotated B. subtilis 168 genome generated by the SEED, one of the most up-to-date and accurate annotations of B. subtilis 168 available. RESULTS: The iBsu1103 model includes 1,437 reactions associated with 1,103 genes, making it the most complete model of B. subtilis available. The model also includes Gibbs free energy change (DeltarG' degrees ) values for 1,403 (97%) of the model reactions estimated by using the group contribution method. These data were used with an improved reaction reversibility prediction method to identify 653 (45%) irreversible reactions in the model. The model was validated against an experimental dataset consisting of 1,500 distinct conditions and was optimized by using an improved model optimization method to increase model accuracy from 89.7% to 93.1%. CONCLUSIONS: Basing the iBsu1103 model on the annotations generated by the SEED significantly improved the model completeness and accuracy compared with the most recent previously published model. The enhanced accuracy of the iBsu1103 model also demonstrates the efficacy of the improved reaction directionality prediction method in accurately identifying irreversible reactions in the B. subtilis metabolism. The proposed improved model optimization methodology was also demonstrated to be effective in minimally adjusting model content to improve model accuracy. PMID- 19555511 TI - Penetrating injury to the chest by an attenuated energy projectile: a case report and literature review of thoracic injuries caused by "less-lethal" munitions. AB - We present the case of a patient who sustained a penetrating injury to the chest caused by an attenuated energy rubber bullet and review the literature on thoracic injuries caused by plastic and rubber "less-lethal" munitions. The patient of this report underwent a right thoracotomy to extract the projectile as well as a wedge resection of the injured lung parenchyma. This case demonstrates that even supposedly safe riot control munition fired at close range, at the torso, can provoke serious injury. Therefore a thorough investigation and close clinical supervision are justified. PMID- 19555512 TI - Localization of CD26/DPPIV in nucleus and its nuclear translocation enhanced by anti-CD26 monoclonal antibody with anti-tumor effect. AB - BACKGROUND: CD26 is a type II, cell surface glycoprotein known as dipeptidyl peptidase (DPP) IV. Previous studies have revealed CD26 expression in T cell leukemia/lymphoma and malignant mesothelioma, and an inhibitory effect of anti CD26 monoclonal antibody (mAb) against the growth of CD26+ cancer cells in vitro and in vivo. The function of CD26 in tumor development is unknown and the machinery with which the CD26 mAb induces its anti-tumor effect remains uncharacterized. RESULTS: The localization of CD26 in the nucleus of T cell leukemia/lymphoma cells and mesothelioma cells was shown by biochemical and immuno-electron microscopic analysis. The DPPIV enzyme activity was revealed in the nuclear fraction of T cell leukemia/lymphoma cells. These expressions of intra-nuclear CD26 were augmented by treatment with the CD26 mAb, 1F7, with anti tumor effect against the CD26+ T cell leukemia/lymphoma cells. In contrast, the CD26 mAb, 5F8, without anti-tumor effect, did not augment CD26 expressions in the nucleus. Biotin-labeled, cell surface CD26 translocated into the nucleus constantly, and this translocation was enhanced with 1F7 treatment but not with 5F8. CONCLUSION: These results indicate that the intra-nuclear CD26 which moves from plasma membrane may play certain roles in cell growth of human cancer cells. PMID- 19555513 TI - Phylogenetic support values are not necessarily informative: the case of the Serialia hypothesis (a mollusk phylogeny). AB - BACKGROUND: Molecular phylogenies are being published increasingly and many biologists rely on the most recent topologies. However, different phylogenetic trees often contain conflicting results and contradict significant background data. Not knowing how reliable traditional knowledge is, a crucial question concerns the quality of newly produced molecular data. The information content of DNA alignments is rarely discussed, as quality statements are mostly restricted to the statistical support of clades. Here we present a case study of a recently published mollusk phylogeny that contains surprising groupings, based on five genes and 108 species, and we apply new or rarely used tools for the analysis of the information content of alignments and for the filtering of noise (masking of random-like alignment regions, split decomposition, phylogenetic networks, quartet mapping). RESULTS: The data are very fragmentary and contain contaminations. We show that that signal-like patterns in the data set are conflicting and partly not distinct and that the reported strong support for a "rather surprising result" (monoplacophorans and chitons form a monophylum Serialia) does not exist at the level of primary homologies. Split-decomposition, quartet mapping and neighbornet analyses reveal conflicting nucleotide patterns and lack of distinct phylogenetic signal for the deeper phylogeny of mollusks. CONCLUSION: Even though currently a majority of molecular phylogenies are being justified with reference to the 'statistical' support of clades in tree topologies, this confidence seems to be unfounded. Contradictions between phylogenies based on different analyses are already a strong indication of unnoticed pitfalls. The use of tree-independent tools for exploratory analyses of data quality is highly recommended. Concerning the new mollusk phylogeny more convincing evidence is needed. PMID- 19555514 TI - New quality and quantity indices in science (NewQIS): the study protocol of an international project. AB - Benchmarking systems are important features for the implementation of efficacy in basic and applied sciences. These systems are urgently needed for many fields of science since there is an imbalance present between funding policies and research evaluation. Here, a new approach is presented with an international study project that uses visualisation techniques for benchmarking processes. The project is entitled New Quality and Quantity Indices in Science (NewQIS). The juxtaposition of classical scientometric tools and novel visualisation techniques can be used to assess quality and quantity in science. In specific, the tools can be used to assess quality and quantity of research activity for distinct areas of science, for single institutions, for countries, for single time periods, or for single scientists. Also, NewQIS may be used to compare different fields, institutions, countries, or scientists for their scientific output. Thus, decision making for funding allocation can be made more transparent. Since governmental bodies that supervise funding policies and allocation processes are often not equipped with an in depth expertise in this area, special attention is given to data visualisation techniques that allow to visualize mapping of research activity and quality. PMID- 19555516 TI - Editorial. Finding suitable reviewers. PMID- 19555517 TI - The impact of garlic on lipid parameters: a systematic review and meta-analysis. AB - In order to determine the impact of garlic on total cholesterol (TC), TAG levels, as well as LDL and HDL, and establish if any variables have an impact on the magnitude of this effect, a meta-analysis was conducted. A systematic literature search of MEDLINE, CINAHL and the Cochrane Database from the earliest possible date through to November 2007 was conducted to identify randomised, placebo controlled trials of garlic that reported effects on TC, TAG concentrations, LDL or HDL. The weighted mean difference of the change from baseline (with 95 % CI) was calculated as the difference between the means in the garlic groups and the control groups using a random-effects model. Subgroup and sensitivity analyses were performed to determine the effects on type, brand and duration of garlic therapy as well as baseline TC and TAG levels, the use of dietary modification, and study quality on the meta-analysis's conclusions. Twenty-nine trials were included in the analysis. Upon meta-analysis garlic was found to significantly reduce TC ( - 0.19; 95 % CI - 0.33, - 0.06 mmol/l) and TAG ( - 0.11; 95 % CI - 0.19, - 0.06 mmol/l) but exhibited no significant effect on LDL or HDL. There was a moderate degree of statistical heterogeneity for the TC and TAG analyses. Garlic reduces TC to a modest extent, an effect driven mostly by the modest reductions in TAG, without appreciable LDL lowering or HDL elevation. Higher baseline line TC levels and the use of dietary modification may alter the effect of garlic on these parameters. Future studies should be conducted evaluating the impact of adjunctive garlic therapy with fibrates or statins on TAG concentrations. PMID- 19555518 TI - Stroke: roles of B vitamins, homocysteine and antioxidants. AB - In the present review concerning stroke, we evaluate the roles of B vitamins, homocysteine and antioxidant vitamins. Stroke is a leading cause of death in developed countries. However, current therapeutic strategies for stroke have been largely unsuccessful. Several studies have reported important benefits on reducing the risk of stroke and improving the post-stroke-associated functional declines in patients who ate foods rich in micronutrients, including B vitamins and antioxidant vitamins E and C. Folic acid, vitamin B6 and vitamin B12 are all cofactors in homocysteine metabolism. Growing interest has been paid to hyperhomocysteinaemia as a risk factor for CVD. Hyperhomocysteinaemia has been linked to inadequate intake of vitamins, particularly to B-group vitamins and therefore may be amenable to nutritional intervention. Hence, poor dietary intake of folate, vitamin B6 and vitamin B12 are associated with increased risk of stroke. Elevated consumption of fruits and vegetables appears to protect against stroke. Antioxidant nutrients have important roles in cell function and have been implicated in processes associated with ageing, including vascular, inflammatory and neurological damage. Plasma vitamin E and C concentrations may serve as a biological marker of lifestyle or other factors associated with reduced stroke risk and may be useful in identifying those at high risk of stroke. After reviewing the observational and intervention studies, there is an incomplete understanding of mechanisms and some conflicting findings; therefore the available evidence is insufficient to recommend the routine use of B vitamins, vitamin E and vitamin C for the prevention of stroke. A better understanding of mechanisms, along with well-designed controlled clinical trials will allow further progress in this area. PMID- 19555515 TI - Emerging strategies for cell and gene therapy of the muscular dystrophies. AB - The muscular dystrophies are a heterogeneous group of over 40 disorders that are characterised by muscle weakness and wasting. The most common are Duchenne muscular dystrophy and Becker muscular dystrophy, which result from mutations within the gene encoding dystrophin; myotonic dystrophy type 1, which results from an expanded trinucleotide repeat in the myotonic dystrophy protein kinase gene; and facioscapulohumeral dystrophy, which is associated with contractions in the subtelomeric region of human chromosome 1. Currently the only treatments involve clinical management of symptoms, although several promising experimental strategies are emerging. These include gene therapy using adeno-associated viral, lentiviral and adenoviral vectors and nonviral vectors, such as plasmid DNA. Exon skipping and cell-based therapies have also shown promise in the effective treatment and regeneration of dystrophic muscle. The availability of numerous animal models for Duchenne muscular dystrophy has enabled extensive testing of a wide range of therapeutic approaches for this type of disorder. Consequently, we focus here on the therapeutic developments for Duchenne muscular dystrophy as a model of the types of approaches being considered for various types of dystrophy. We discuss the advantages and limitations of each therapeutic strategy, as well as prospects and recent successes in the context of future clinical applications. PMID- 19555519 TI - Vitamin D: emerging new roles in insulin sensitivity. AB - The growing incidence of prediabetes and clinical type 2 diabetes, in part characterised by insulin resistance, is a critical health problem with consequent devastating personal and health-care costs. Vitamin D status, assessed by serum 25-hydroxyvitamin D levels, is inversely associated with diabetes in epidemiological studies. Several clinical intervention studies also support that vitamin D, or its active metabolite 1,25-dihydroxyvitamin D (1,25(OH)2D), improves insulin sensitivity, even in subjects with glucose metabolism parameters classified within normal ranges. The mechanisms proposed which may underlie this effect include potential relationships with improvements in lean mass, regulation of insulin release, altered insulin receptor expression and specific effects on insulin action. These actions may be mediated by systemic or local production of 1,25(OH)2D or by suppression of parathyroid hormone, which may function to negatively affect insulin sensitivity. Thus, substantial evidence supports a relationship between vitamin D status and insulin sensitivity; however, the underlying mechanisms require further exploration. PMID- 19555520 TI - A review and meta-analysis of the effect of weight loss on all-cause mortality risk. AB - Overweight and obesity are associated with increased morbidity and mortality, although the range of body weights that is optimal for health is controversial. It is less clear whether weight loss benefits longevity and hence whether weight reduction is justified as a prime goal for all individuals who are overweight (normally defined as BMI>25 kg/m2). The purpose of the present review was to examine the evidence base for recommending weight loss by diet and lifestyle change as a means of prolonging life. An electronic search identified twenty-six eligible prospective studies that monitored subsequent mortality risk following weight loss by lifestyle change, published up to 2008. Data were extracted and further analysed by meta-analysis, giving particular attention to the influence of confounders. Moderator variables such as reason for weight loss (intentional, unintentional), baseline health status (healthy, unhealthy), baseline BMI (normal, overweight, obese), method used to estimate weight loss (measured weight loss, reported weight loss) and whether models adjusted for physical activity (adjusted data, unadjusted data) were used to classify subgroups for separate analysis. Intentional weight loss per se had a neutral effect on all-cause mortality (relative risk (RR) 1.01; P = 0.89), while weight loss which was unintentional or ill-defined was associated with excess risk of 22 to 39 %. Intentional weight loss had a small benefit for individuals classified as unhealthy (with obesity-related risk factors) (RR 0.87 (95 % CI 0.77, 0.99); P = 0.028), especially unhealthy obese (RR 0.84 (95 % CI 0.73, 0.97); P = 0.018), but appeared to be associated with slightly increased mortality for healthy individuals (RR 1.11 (95 % CI 1.00, 1.22); P = 0.05), and for those who were overweight but not obese (RR 1.09 (95 % CI 1.02, 1.17); P = 0.008). There was no evidence for weight loss conferring either benefit or risk among healthy obese. In conclusion, the available evidence does not support solely advising overweight or obese individuals who are otherwise healthy to lose weight as a means of prolonging life. Other aspects of a healthy lifestyle, especially exercise and dietary quality, should be considered. However, well-designed intervention studies are needed clearly to disentangle the influence of physical activity, diet strategy and body composition, in order to define appropriate advice to those populations that might be expected to benefit. PMID- 19555521 TI - Joint BAPEN and British Society of Gastroenterology Symposium on 'Coeliac disease: basics and controversies'. Coeliac disease: optimising the management of patients with persisting symptoms? AB - The vast majority of patients with coeliac disease will derive benefit from a gluten-free diet. However, some patients will not improve on the gluten-free diet or they will have a relapse of their symptoms. The present review will focus on this group of patients. Definitions for non-responsive coeliac disease and refractory coeliac disease will be provided. The most common reason for recurrent symptoms is continued gluten exposure. Other causes of persisting symptoms are discussed, including alternative causes of villous atrophy or co-existent pathology. Current literature is reviewed, including an initial investigation strategy for patients with persisting symptoms. A pragmatic management plan is described that can be initiated by any clinician. Finally, the current optimal investigational pathway for patients with refractory (or suspected refractory) coeliac disease is discussed and the reported effects of a number of therapeutic options are summarised. The aim of the present article is to provide clinicians with an up-to-date review of the literature in this clinical field and allow them to determine the most appropriate management strategy. PMID- 19555522 TI - Molecular epidemiology of African sleeping sickness. AB - Human sleeping sickness in Africa, caused by Trypanosoma brucei spp. raises a number of questions. Despite the widespread distribution of the tsetse vectors and animal trypanosomiasis, human disease is only found in discrete foci which periodically give rise to epidemics followed by periods of endemicity A key to unravelling this puzzle is a detailed knowledge of the aetiological agents responsible for different patterns of disease--knowledge that is difficult to achieve using traditional microscopy. The science of molecular epidemiology has developed a range of tools which have enabled us to accurately identify taxonomic groups at all levels (species, subspecies, populations, strains and isolates). Using these tools, we can now investigate the genetic interactions within and between populations of Trypanosoma brucei and gain an understanding of the distinction between human- and nonhuman-infective subspecies. In this review, we discuss the development of these tools, their advantages and disadvantages and describe how they have been used to understand parasite genetic diversity, the origin of epidemics, the role of reservoir hosts and the population structure. Using the specific case of T.b. rhodesiense in Uganda, we illustrate how molecular epidemiology has enabled us to construct a more detailed understanding of the origins, generation and dynamics of sleeping sickness epidemics. PMID- 19555523 TI - Social problem solving, autobiographical memory, trauma, and depression in women with borderline personality disorder and a history of suicide attempts. AB - OBJECTIVES: The primary aim of this study was to compare the retrieval of autobiographical memory and the social problem-solving performance of individuals with borderline personality disorder (BPD) and a history of suicide attempts, with and without concurrent diagnoses of depression and/or post-traumatic stress disorder (PTSD), to that of controls. Additionally, the relationships between autobiographical memory, social problem-solving skills, and various clinical characteristics were examined in the BPD group. DESIGN: Individuals with BPD who had made at least two suicide attempts were compared to controls with regard to specificity of autobiographical memory and social problem-solving skills. Autobiographical memory specificity and social problem-solving skills were further studied in the BPD group by comparing depressed participants to non depressed participants; and autobiographical memory specificity was also studied by comparing participants with and without PTSD. METHOD: A total of 47 women with a diagnosis of BPD and 30 controls completed the Autobiographical Memory Test, assessing memory specificity, and the means-end problem solving-procedure, measuring social problem-solving skills. The prevalence of suicidal/self injurious behaviour, and the exposure to violence, was also assessed in the BPD group. RESULTS: Compared to controls, participants with BPD showed reduced specificity of autobiographical memory, irrespective of either concurrent depression, previous depression, or concurrent PTSD. The depressed BPD group displayed poor problem-solving skills. Further, an association between unspecific memory and poor problem-solving was displayed in the BPD group. CONCLUSION: Our results confirmed that reduced specificity of autobiographical memory is an important characteristic of BPD individuals with a history of suicide attempt, independent of depression, or PTSD. Reduced specificity of autobiographical memory was further related to poor social problem-solving capacity in the BPD group. PMID- 19555525 TI - Low-income countries: where are we going with TB-HIV in 2009? PMID- 19555524 TI - The effects of social desirability response bias on STAXI-2 profiles in a clinical forensic sample. AB - OBJECTIVES: This study investigated the proposition that the 'State-trait anger expression inventory' (2nd ed.; STAXI-2) is susceptible to impression management (IM) and Self-Deceptive Enhancement (SDE) in clinical forensic populations. It was hypothesized that individuals engaging in IM would report significantly lower levels of trait anger, external expression of anger, and internal expression of anger on the STAXI-2. Those reporting above average SDE were predicted to claim higher levels of anger control. DESIGN: A between-groups design was used, comparing STAXI-2 scores of individuals who reported high levels of IM and SDE to those who did not. METHOD: One-hundred and fifty-nine male patients of a community forensic mental health service, referred for assessment of stalking behaviours, completed the STAXI-2 and Paulhus Deception Scales (PDS). Individuals engaging in high levels of IM and SDE were compared to low scorers in regard to STAXI-2 scales using Mann-Whitney U tests. RESULTS: Individuals engaging in IM had significantly lower levels of reported trait anger, outward expression of anger, and inward expression of anger, and higher levels of anger control. Similar results were found with the SDE scale, although the magnitude of the effect was smaller and not apparent on all subscales. CONCLUSIONS: The STAXI-2 was vulnerable to social desirability response bias in this sample of forensic clients. Where the STAXI-2 is used as a basis for treatment recommendations and decisions, it should be administered and interpreted in conjunction with a recognized measure of such bias to improve validity. PMID- 19555526 TI - Scaling up tuberculosis culture services: a precautionary note. PMID- 19555527 TI - Registering initial defaulters and reporting on their treatment outcomes. AB - This Unresolved Issues article highlights three original articles that appeared last year in the Journal discussing the phenomenon of initial defaulters. There are three important challenges with patients that appear in the laboratory sputum register but are not recorded in the tuberculosis (TB) patient register: the first is how to identify these patients, trace them and get them on to treatment as soon as possible; the second is how to register and report on these cases as part of the case-finding component of TB control; and the third is whether to include these initial default patients in the cohort analysis of treatment outcomes. We recommend a step-wise approach to these challenges and advocate that these patients be included, wherever possible, in the TB patient register and in the cohort analysis of treatment outcomes. PMID- 19555528 TI - Health systems that serve: what's the bottom line? AB - SETTING: Health services in low- and middle-income countries. BACKGROUND: The Global Plan to Stop TB, 2006-2015. OBJECTIVE: Using a framework for evaluation of public health systems, to evaluate evidence that tuberculosis (TB) services contribute to strengthening the health systems. DESIGN: Critical evaluation of published material. RESULTS: The Global Plan to Stop TB 2006-2015 identifies strengthening the health systems as one of its components. Published material illustrates substantial improvement of quality of TB services over the past decade. However, even where these services have achieved a high level of quality, there is little evidence to indicate that other health services in the same locations show similar quality. CONCLUSION: Policies, strategies and actions to strengthen health systems through TB services will require specific plans and priorities to achieve their objectives; this will not occur as a natural effect of improving TB services. PMID- 19555529 TI - Rifapentine vs. rifampicin for the treatment of pulmonary tuberculosis: a systematic review. AB - OBJECTIVE: To evaluate the efficacy and safety of rifapentine (RPT) vs. rifampicin (RMP) for the treatment of pulmonary tuberculosis (PTB). DESIGN: Systematic review of randomised controlled trials (RCTs) that compared combination drug regimens containing RPT with those containing RMP for the treatment of drug-susceptible or previously untreated PTB. RESULTS: Nine RCTs were identified. Statistically significant differences were not found in the cure rates, severe adverse effects, severe hepatotoxicity or bacteriological relapse rates between the regimens containing once- or twice-weekly RPT and those containing daily RMP for human immunodeficiency virus (HIV) negative patients, but were found in the bacteriological relapse rates between regimens containing once-weekly or less frequent RPT and those containing twice- or thrice-weekly RMP: the pooled relative risks in the two subgroups were respectively 1.71 (95%CI 1.13-2.58, P = 0.01) and 2.44 (95%CI 1.15-5.18, P = 0.02). The trial for HIV positive patients did not show significant differences in the sputum conversion rate, severe adverse effects or bacteriological relapse rate between the RPT- and RMP-containing regimens; four of the five relapses were associated with the RPT containing regimen, but none of the three relapses with the RMP-containing regimen produced monoresistance to rifamycin (RIF). CONCLUSION: Once- or twice weekly RPT and daily RMP have similar efficacy and safety for the treatment of HIV-negative PTB, but once-weekly or less frequent use of RPT, in comparison with twice- or thrice-weekly RMP, increases the risk of bacteriological relapse. RPT might increase the risk of resistance to RIF in HIV-positive patients. PMID- 19555530 TI - Interferon-gamma release assays in immigrant contacts and effect of remote exposure to Mycobacterium tuberculosis. AB - OBJECTIVE: To assess the association between remote exposure to tuberculosis (TB) and results of the tuberculin skin test (TST), and two interferon-gamma release assays (IGRAs)-QuantiFERON-TB Gold In-Tube (QFT-GIT) and T-SPOT.TB-in immigrant contacts of sputum smear-positive TB patients. METHODS: Immigrants aged >or=16 years in close contact with smear-positive TB patients were included. QFT-GIT and T-SPOT.TB were performed if the TST induration size was >or=5 mm. Associations between test results and origin from an endemic country were assessed. RESULTS: Of 433 close contacts, 322 (74%) had TST >or=5 mm, of whom, 282 (88%) had valid test results for all assays. Positive QFT-GIT results were obtained for 152/282 (54%) and positive T-SPOT.TB for 168/282 (60%). After adjustment for age, sex and recent contact, positive IGRA results and TST results >/=10 mm were found to be more frequent among immigrants who originated from Africa, in particular sub Saharan Africa. CONCLUSION: When IGRAs are used to determine latent TB infection in foreign-born individuals, positive findings not only relate to recent TB infection, but also reflect prior TB exposure in the country of origin. This late reactivity will limit their usefulness in contact investigations among immigrants originating from endemic areas. PMID- 19555531 TI - Factors associated with positive tuberculin skin test results among HIV-infected persons in Orel Oblast, Russia. AB - BACKGROUND: The treatment of persons living with human immunodeficiency virus/acquired immune-deficiency syndrome (PLWHAs) for latent tuberculosis infection (LTBI) reduces tuberculosis (TB) morbidity. Despite a high TB burden and an expanding human immunodeficiency virus epidemic, Russia had limited data on the utility of the tuberculin skin test (TST) for LTBI diagnosis in PLWHAs. OBJECTIVE: To determine the prevalence and predictors of positive TSTs in PLWHAs in Orel Oblast. METHODS: A total of 150 consenting PLWHAs being followed up at the AIDS Center were administered a TST and a questionnaire for risk factors for LTBI. A positive TST result was defined as >or=5 mm induration. RESULTS: Of the 150 subjects, 67% were male and 74% were aged <30 years. Of the PLWHAs tested, 26% had a positive TST result, while among PLWHAs with CD4(+) >500 cells/ml, 36% were TST-positive. TST positivity varied inversely with CD4(+) cell count. Among PLWHAs with a history of injection drug use, the primary risk factor for HIV, 29 (31.9%) were positive. CONCLUSIONS: A high proportion of tested PLWHAs had a positive TST and could benefit from preventive therapy (PT) to reduce the risk of TB. A TB control programme in Russia should therefore include TST screening among PLWHAs and PT, besides active TB case finding and treatment. PMID- 19555532 TI - Comparison of Lumin LED fluorescent attachment, fluorescent microscopy and Ziehl Neelsen for AFB diagnosis. AB - DESIGN: Cross-sectional studies in Russia (n = 502) and Macedonia (n = 205), with fluorochrome-stained sputum examined by 1) the new Lumin light emitting diode (LED) fluorescent attachment on a light microscope, and 2) conventional fluorescent microscope (CFM) available in each laboratory, and compared to 3) Ziehl-Neelsen (ZN) restaining/reading of the same smears. Poor readings of ZN restained smears in Russia stimulated a retrospective laboratory registry analysis for sensitivity and specificity of directly ZN-stained smears (n = 791) from a previous period. RESULTS: In Macedonia, the sensitivity of the Lumin and CFM were 87.8%, and that of restained ZN smears with conventional light microscope was 78.0%. In Russia, sensitivity was as follows: Lumin 72.8%, CFM 52.5%; re-stained ZN smears 28.5% and directly ZN stained smears 55.6%. CONCLUSION: Fluorescence microscopy is more sensitive than conventional microscopy. The Lumin attachment to conventional light microscopes provided results equal to or better than the CFMs. Smear restaining for ZN showed a 12% advantage for Lumin and CFM in Macedonia, in line with other meta-analyses. Restaining for ZN gave poor results in Russia for unknown reasons. Retrospective analysis of directly ZN-stained smears showed 55.6% sensitivity compared to the Lumin (72.8%), which is also in line with the superiority of fluorescent microscopy reported in literature. PMID- 19555533 TI - Two vs. three sputum samples for microscopic detection of tuberculosis in a high HIV prevalence population. AB - SETTING: A busy urban hospital in Cameroon. OBJECTIVES: To compare the yield in bacteriologically proven tuberculosis (TB) cases examining two morning vs. three spot-morning-spot sputum specimens (MM vs. SMS) by direct microscopy for acid fast bacilli (AFB). DESIGN: Repeated temporal cross-over between MM and SMS sampling for successive TB suspects, using culture as gold standard. RESULTS: A total of 799 suspects were screened using the MM strategy, identifying 223 smear positives, and 808 suspects with the SMS strategy, yielding 236 smear-positives. Of the MM, 256 were culture-positive, of whom 195 (76%) were smear-positive. For SMS, these figures were respectively 281 and 206 (73%), a non-significant difference. The MM and SMS strategies also detected respectively 28 and 30 smear positive cases not confirmed by culture. No cases were lost to treatment with either strategy. CONCLUSIONS: In this population with a high prevalence of human immunodeficiency virus (HIV) with late case presentation, smear microscopy of two morning specimens detected at least as many positive cases as the classical strategy, and no cases were lost before treatment. Two specimens for initial TB suspect screening can thus be recommended, also without excessive workload. Comparative studies in populations presenting with paucibacillary sputum are needed to determine the equivalent quality and yield of an alternative strategy with two spot specimens at consultation. PMID- 19555534 TI - Evaluation of a multi-antigen test based on B-cell epitope peptides for the serodiagnosis of pulmonary tuberculosis. AB - SETTING: Two sample panels: 1) 20 pulmonary tuberculosis (PTB) patients and 10 healthy subjects from a country with a low incidence of TB (Italy); and 2) 47 PTB patients and 26 healthy subjects from a country with a high incidence of TB (Morocco). OBJECTIVE: To identify a combination of Mycobacterium tuberculosis peptides useful for the serodiagnosis of active PTB. METHODS: Fifty-seven B-cell epitope peptides of M. tuberculosis were evaluated by immunoenzymatic assay and the data were analysed using logistic regression analysis and the random forest method. RESULTS: The best discriminating peptide between PTB patients and healthy subjects from the sample of the low TB incidence country was the 23 amino acid peptide of the Rv3878 protein. The sensitivity and specificity were respectively 65% and 100%. The same peptide had a sensitivity and specificity of respectively 47% and 100% for the sample from the high TB incidence country. The best combination of peptides was a pool of nine peptides which had a sensitivity of 70.2% and a specificity of 100% in the high TB incidence country. CONCLUSIONS: The 9-peptide pool can be useful in identifying patients with active PTB. PMID- 19555535 TI - Improved early results for patients with extensively drug-resistant tuberculosis and HIV in South Africa. AB - SETTING: A public tuberculosis (TB) referral hospital in KwaZulu-Natal, South Africa. OBJECTIVE: To present treatment outcomes of patients with extensively drug-resistant tuberculosis (XDR-TB) patients and human immunodeficiency virus (HIV) coinfection with and without highly active antiretroviral therapy. METHODS: Retrospective cohort study. Eligible patients had drug susceptibility testing that met a consensus definition for XDR-TB, and agreed to treatment. Therapy was based on drug susceptibilities, available medications and patient tolerance. RESULTS: Overall, 60 XDR-TB patients initiated therapy with a median number of 5.5 drugs. Of these, 43 (72%) were HIV-positive, and 21 (49%) were on antiretroviral therapy; 29 HIV-infected patients (67%) had available CD4 counts, with a median CD4 count of 200.5 cells/mm(3) (standard deviation 127.4 cells/mm(3)). Of 60 patients, 31 (52%) had adverse events (AEs), and 17/60 patients (28%) had severe AEs. During follow-up, 12/60 (20%) experienced sputum culture conversion, while 25/60 (42%) patients died. None of the following was significantly associated with mortality: HIV status, previous MDR diagnosis or severe AEs. DISCUSSION: In this study, it was possible to treat HIV-XDR-TB coinfected patients and prolong survival in a resource-limited setting. We highlight the challenges in treatment, including high frequencies of AEs and death. Expanded identification of cases, prompt referral for treatment, and attention to management of comorbidities may facilitate successful treatment of XDR-TB in HIV-infected patients. PMID- 19555536 TI - HAART and risk of tuberculosis in HIV-infected South African children: a multi site retrospective cohort. AB - SETTING: Four human immunodeficiency virus (HIV) clinics located at South African tertiary hospitals. OBJECTIVE: To assess the effectiveness of highly active antiretroviral therapy (HAART) in reducing incident tuberculosis (TB) in HIV infected children. DESIGN: Retrospective cohort. RESULTS: A total of 1132 children's records were included in the study. At entry to the cohort, the median (interquartile range [IQR]) age, CD4%, CD4 count and viral load of all children was respectively 6.3 years (4.1-8.8), 15% (9.0-22.2), 576 cells/mm(3) (287-960) and 160 000 copies/ml (54 941.5-449 683); 75.9% were started on HAART. The male:female ratio was 1:1, and median follow-up time was 1.7 years. In children whose follow-up included both pre-HAART and on-HAART periods, the incidence of clinically diagnosed TB was respectively 21.1 per 100 person-years (py; 95%CI 18.2-24.4) and 6.4/100 py (95%CI 4.8-8.1), and when restricted to confirmed cases, respectively 3.1/100 py (95%CI 2.2-4.2) and 0.8/100 py (95%CI 0.5-1.4). Only 23% of all cases of TB were microbiologically confirmed. Multivariate analyses showed that HAART reduced incident TB by approximately 70%, both for confirmed and all TB cases. CONCLUSIONS: In this high TB burden country, the incidence of diagnosis of TB in HIV-infected children is at least as high as that of adults. HAART reduces incident TB, but further prospective TB preventive and diagnostic studies are urgently needed in children. PMID- 19555537 TI - High prevalence of Mycobacterium tuberculosis infection and disease in children and adolescents with type 1 diabetes mellitus. AB - SETTING: Western Cape Province, South Africa. OBJECTIVES: To describe the prevalence of tuberculosis (TB) infection and disease in children with type 1 diabetes and to investigate the association between glycaemic control and prevalence of TB infection and disease. DESIGN: Cross-sectional hospital-based study conducted at two public referral hospitals. All children and adolescents (aged <21 years) with type 1 diabetes underwent a Mantoux tuberculin skin test (>or=10 mm classified as Mycobacterium tuberculosis infection), measurement of glycosylated haemoglobin and a chest radiograph. Patients with symptoms suggestive of TB were investigated using mycobacterial culture. Radiologically and/or bacteriologically confirmed disease was classified as TB disease. RESULTS: Of 291 eligible patients, 258 (88.7%) were included (58% female). The prevalence of M. tuberculosis infection was 29.8% (95%CI 24.2-35.4); nine patients were diagnosed with prevalent TB disease (point prevalence disease 3488 per 100,000 population). Poor glycaemic control (hazard ratio 1.39, 95%CI 1.18-1.63 per unit increase in glycated haemoglobin [HbA1c]) and contact with a TB source case (P = 0.0011) was associated with prevalent TB disease. CONCLUSIONS: There is a high prevalence of TB disease in diabetic children and adolescents in this setting. Routine TB screening of children with type 1 diabetes may be indicated in settings highly endemic for TB. Preventive treatment should be considered for diabetic children with proof of TB exposure and/or infection. PMID- 19555538 TI - Management of life-threatening haemoptysis in an area of high tuberculosis incidence. AB - BACKGROUND: Emergency admissions with life-threatening haemoptysis in an area of high tuberculosis (TB) incidence at the University of Stellenbosch and Tygerberg Academic Hospital, South Africa. It is unclear if lung resection is regularly indicated to prevent recurrence following bronchial artery embolisation (BAE). OBJECTIVE: To prospectively evaluate risk factors for recurrence as selection criteria for surgery following embolisation: lack of complete cessation of haemoptysis, need for blood transfusion, presence of aspergilloma and absence of active TB. DESIGN: Prospective interventional study with 1-year follow-up. RESULTS: Within a 7-month period, 101 consecutive patients were admitted. Seven were excluded and 12 died shortly after admission. Haemoptysis ceased on medical treatment alone within 24 h in 21 of the remaining 82 patients. Their 1-year mortality was 10%. Eleven of 61 patients referred for emergency embolisation died before discharge. Of the 50 patients remaining at risk of recurrence, 38 (76%) were at low risk and 12 (24%) at high risk. Five of these patients (10% of those at risk) underwent surgery. Patients at low risk and operated patients had an uneventful course over 1 year, but two deaths occurred among the seven inoperable patients at high risk. CONCLUSION: Lung resection surgery following successful BAE for life-threatening haemoptysis can safely be avoided in patients at low risk of recurrence. PMID- 19555539 TI - Management and treatment outcomes of tuberculous patients, eastern Paris, France, 2004. AB - BACKGROUND: France was one of the few European countries without a national tuberculosis (TB) treatment outcome monitoring system until 2007. OBJECTIVE: To examine TB management and treatment outcomes in the eastern Paris region, and to identify patient- and management-dependent factors affecting treatment outcome. METHODS: This retrospective study focused on all cases of microbiologically confirmed Mycobacterium tuberculosis cases diagnosed in 2004 in the eastern Paris region, one of the areas of France with the highest frequency of TB. RESULTS: Treatment outcomes of 629 identified cases (males 69.6%, median age 37 years, socio-economically disadvantaged 44%, foreign-born 78%) were as follows: treatment success 70.1% (95%CI 66.5-73.7), treatment interruption 4.9% (95%CI 3.2 6.6), loss to follow-up 15.0% (95%CI 12.2-17.8), death 5.7% (95%CI 3.9-7.9), transfers 4.3% (95%CI 3.5-5.1). Non-completion of treatment was associated with sputum smear positivity, injection drug use, non-adherence and irregular follow up in univariate analysis, and with irregular follow-up and non-adherence in multivariate analysis. Duration of TB treatment and follow-up medical visits were not applied as recommended in more than a third of cases. CONCLUSION: The treatment success rate observed in this study (70.1%) is below the World Health Organization target of 85%, and requires the implementation and evaluation of interventions to increase treatment success rates. PMID- 19555540 TI - Tuberculosis services and treatment outcomes in private and public health care facilities in Thailand, 2004-2006. AB - BACKGROUND: The World Health Organization recommends that national tuberculosis (TB) programs encourage public and private providers to follow the 'International standards for tuberculosis care'. We assessed services and treatment outcomes in TB patients in public and private facilities to inform public-private mix scale up in Thailand. METHODS: We prospectively collected data on TB patients in four provinces and the national infectious diseases hospital during 2004-2006. We analyzed services and outcomes among new pulmonary TB patients according to facility type. RESULTS: Of 7526 patients, 4539 (60%) were treated in small public facilities, 2275 (30%) in large public facilities and 712 (10%) in private facilities. Compared with the private sector, more public sector patients had at least two sputum smears examined, were prescribed a standard anti-tuberculosis regimen and received directly observed therapy; however, public sector facilities also performed suboptimally. Treatment outcomes were unsuccessful for 237 (33%) patients in private facilities, and for respectively 1018 (23%) and 655 (29%) patients in small and large public facilities. CONCLUSIONS: TB diagnostic and treatment services and outcomes should be enhanced in both public and private facilities in Thailand. Initiatives are needed to improve treatment outcomes and increase the use of microscopy, standardized TB regimens, and directly observed therapy in the public and private sectors. PMID- 19555541 TI - Factors influencing tuberculosis management in rural south-west Kenya. AB - SETTING: The Abagusii people in rural south-west Kenya. OBJECTIVE: To investigate tuberculosis (TB) treatment pathways and factors associated with treatment seeking behaviour. METHOD: Quantitative data were obtained using a structured questionnaire administered to 100 household heads, while qualitative data were obtained from three focus group discussions (FGDs), six key informant interviews and seven case histories that focused on experiences of TB. RESULTS: Data reveal that patients follow oscillatory pathways in treatment seeking. Treatment-seeking behaviour is influenced by socio-structural and superstructural forces such as beliefs and perceptions regarding treatment and disease-causing factors. CONCLUSION: To improve TB treatment, these socio-structural and superstructural factors should be addressed. PMID- 19555542 TI - The Beijing genotype is associated with young age and multidrug-resistant tuberculosis in rural Vietnam. AB - BACKGROUND: Associations between multidrug resistance and the Mycobacterium tuberculosis Beijing genotype have been described mainly in populations with poor tuberculosis (TB) control such as prisons and inner cities, and may reflect shared risk factors rather than a biological association. OBJECTIVE: To study the association between genotype and drug resistance among TB patients in a population with adequate TB control. SETTING: Three rural districts in Vietnam. The study was performed at the Pham Ngoc Thach Tuberculosis and Lung Disease Hospital, Ho Chi Minh City, and the Tien Giang Provincial Tuberculosis and Lung Disease Hospital, My Tho, Vietnam. METHODS: Pretreatment sputum specimens were collected for culture, drug susceptibility testing and spoligotyping of all sputum smear-positive pulmonary TB patients consecutively diagnosed over a 3-year period. RESULTS: Beijing genotype infections were observed in 614 of 1744 (35%) patients. Beijing strains were more common among female (adjusted odds ratio [aOR] 1.4, P = 0.005), young (aOR 2.8, P < 0.001) and previously treated patients (aOR 2.4, P < 0.001). The Beijing genotype was associated with any resistance (aOR 3.7, P < 0.001) and multidrug resistance (aOR 6.8, P < 0.001) among new patients, and with any resistance (aOR 2.7, P = 0.005) but not with multidrug resistance (aOR 1.4, P = 0.545) among previously treated patients. CONCLUSION: In Vietnam, Beijing genotype is associated with young age and in new patients with multidrug resistance despite adequate TB control, suggesting a biological association. This potentially undermines the effectiveness of TB control in countries where Beijing genotype infections are common. PMID- 19555544 TI - Rapid, accurate determination of multidrug resistance in M. tuberculosis isolates and sputum using a biochip system. AB - OBJECTIVE: To develop and evaluate a rapid biochip system for the determination of multidrug-resistant tuberculosis (MDR-TB) in Mycobacterium tuberculosis isolates and clinical sputum samples. DESIGN: We developed a total solution-based system, including a biochip kit, apparatus for sample preparation, hybridisation, washing and data acquisition, and dedicated software for automated diagnosis. The biochip simultaneously identifies M. tuberculosis and detects the most commonly found mutations in the rpoB, katG and inhA genes. The system was assessed with 330 mycobacterial isolates and 129 sputum samples for rifampicin (RMP), and with 205 isolates and 105 sputum samples for isoniazid (INH), and then compared to DNA sequencing and conventional drug susceptibility testing (DST). RESULTS: The entire biochip assay took 6 h. The concordance rate between the biochip assay and the DNA sequencing results was 100%. Compared to conventional DST, the concordance rates were 91.8% for isolates and 94.6% for sputum samples for RMP resistance, and 70.2% for isolates and 78.1% for sputum samples for INH resistance. CONCLUSION: The biochip system provides a simple, rapid, reliable and accurate clinical assay for the parallel detection of M. tuberculosis and prevalent MDR-TB in a 6 h procedure, using either culture isolates or sputum samples for diagnosis. PMID- 19555545 TI - Exacerbation of latent genital tuberculosis during in vitro fertilisation and pregnancy. PMID- 19555543 TI - Radiometric vs. agar proportion methods for assessing ethionamide resistance and its clinical effects in multidrug-resistant tuberculosis. AB - SETTING: Three centres for the treatment of multidrug-resistant tuberculosis (MDR TB) in Istanbul, Turkey: Heybeliada Centre for Chest Diseases and Thoracic Surgery, Sureyyapasa Centre for Chest Diseases and Thoracic Surgery and Yedikule Centre for Chest Diseases and Thoracic Surgery. OBJECTIVE: To evaluate the presence of ethionamide (ETH) resistance and its effect on time to sputum smear negativity in MDR-TB patients who had not previously received second-line anti tuberculosis drugs. DESIGN: Drug susceptibility testing for isoniazid (INH), rifampicin (RMP), ethambutol, streptomycin and ETH was performed on 50 patients treated between August 2004 and May 2005. Indirect agar proportion and BACTEC methods were used to determine ETH susceptibility. RESULTS: Of the patients who were resistant to at least INH and RMP, 11 (22%) (three [27.3%] new and eight [72.7%] retreatment) were resistant to ETH with the BACTEC method. Of 18 new patients, three (16.6%) were ETH-resistant using the BACTEC method compared to 8/32 (25%) retreatment patients. The mean time to smear negativity was 75.2 days in ETH-resistant patients and 50 days in susceptible patients (P < 0.05). Both ETH-resistant and -susceptible groups were homogeneous for factors that may have a possible effect on time to conversion. CONCLUSION: Not only ETH resistance but also age and radiologically advanced disease adversely affected time to sputum conversion. PMID- 19555546 TI - Monolithic stationary phases in classic and chiral pharmaceutical analysis with CEC and pCEC. AB - New stationary phases play an important role in the evolution of separation methods. The use of monolithic phases in capillary columns has become rather widespread. The ease of their preparation, their versatility, and the abundance of available chemistries increase their attractiveness, even dethroning the particulate phases. This review paper overviews the different types of monoliths used in capillary electrochromatography and pressurized capillary electrochromatography for the enantiomeric separation of chiral molecules and the analysis of pharmaceutically relevant molecules. The diverse methods of monolith preparation as well as the different types of used materials are discussed, as well as the ways they can be modified to fulfill given analysis needs (e.g., selectivity, efficiency of separation, speed of analysis, and economical interest). In addition, some of the advantages and drawbacks of the different monolithic materials are mentioned. PMID- 19555547 TI - Recent applications of organic monoliths in capillary liquid chromatographic separation of biomolecules. AB - Monolithic columns are an attractive alternative to traditional particulate solid phases for capillary liquid chromatography. A monolith is a continuous interconnected skeleton with large through-pores. This structure reduces the diffusion path and provides high permeability, resulting in excellent separation efficiency. The integral structure enhances the mechanical strength, while the large through-pores (a few microm) have very low flow impedance. This combination allows smaller diameter monolithic columns to be operated at higher flow-rates, simultaneously increasing both sensitivity and throughput. Polymeric monoliths were first described back in the 1960s, but the first successful ones designed for protein separations appeared much later, in the late 1980s. Organic monoliths are based upon copolymerization of a monofunctional and a bifunctional (uncommonly trifunctional) organic precursor in the presence of a suitable initiator and porogenic solvents. During the last 15 years, a vast number of different monomers and crosslinkers have been introduced and copolymerized using different polymerization techniques and initiators. Various mechanisms, including thermally- and UV-initiated free radical polymerization, as well as ring opening metathesis copolymerizations, have been demonstrated for the preparation of monolithic columns. In this review, we summarize the recent application of different organic monoliths, including styrene-, acrylate-, methacrylate-, and acrylamide for the liquid separation of biomolecules (e.g., proteins, peptides, and oligonucleotides). PMID- 19555548 TI - Monolithic silica stationary phases in liquid chromatography. AB - During the last few decades, monolithic stationary phases (based on silica and polymers) have been used for fast separations in high-performance liquid chromatography (HPLC) and capillary electro-chromatography (CEC). The present article describes the preparation, properties, and applications of these stationary phases. Attempts have been made to discuss the preparation of reversed phase monolithic HPLC columns and CEC capillaries. The chromatographic properties of these phases have been described. The applications included their use in HPLC and CEC modalities of liquid chromatography. The optimization of separations of various molecules on these phases has been discussed. Efforts were also made to predict the future perspectives of monolithic stationary phases. PMID- 19555549 TI - Accelerating the quality control of pharmaceuticals using monolithic stationary phases: a review of recent HPLC applications. AB - High-performance liquid chromatography (HPLC) is the predominant technique in the quality control of pharmaceutical formulations. HPLC offers highly sensitive and selective analytical methods with increased robustness and separation efficiency. The majority of HPLC-based methodologies are based on the usage of particulate based columns and cannot be applied at elevated flow rates (> 2 mL/min) due to excessive back-pressure. Therefore, the typical duration of separation cycles is in the range of 5-30 min, making most assays time-consuming, especially when a lot of samples have to be analyzed in a minimum amount of time. Monolithic stationary phases for HPLC offer a very interesting alternative to conventional particulate-based columns. The size of the skeleton and the distribution of the pores of monolithic materials offer the possibility of developing efficient separation protocols at higher flow-rates due to the low pressure-drop across the column. The present review intends to cover the applications of monolithic based HPLC stationary phases in the quality control of pharmaceutical formulations, including identification of active pharmaceutical ingredient, assay, purity, dissolution, blending, and dosage uniformity, etc. PMID- 19555550 TI - Surface diffusion in C(18)-silica monolithic stationary phase. AB - A peak parking-moment analysis method was used for the measurement of surface diffusion coefficient (D(s)) in a reversed-phase liquid chromatography (RPLC) system consisting of a C(18)-silica monolithic column and a mixture of methanol and water (70/30, v/v). The D(s) values experimentally measured were analyzed by considering the correlation with corresponding values of molecular diffusivity (D(m)) and the retention equilibrium constant (K(a)). It seems that the correlation between D(s)/D(m) and K(a) is represented by a single curve irrespective of the RPLC conditions of temperature and the type of sample compounds. The increase in K(a) is accompanied with the decrease in D(s)/D(m). Oppositely, the ratio of D(s)/D(m) increases and approaches around unity when K(a) infinitely decreases. It seems that surface diffusion is originally similar to molecular diffusion and that it is restricted due to the sample retention. These characteristics of surface diffusion are the same between the C(18)-silica monolithic stationary phase and the conventional C(18)-silica gel particles. In addition, the values of K(a) and D(s) are also comparable between them. It is concluded that basic properties concerning the retention equilibrium and surface diffusion of the C(18)-silica monolithic stationary phase are almost the same as those of the conventional C(18)-silica gel particles in spite of the difference between their structural characteristics. PMID- 19555551 TI - The design by molecular dynamics modeling and simulations of porous polymer adsorbent media immobilized on the throughpore surfaces of polymeric monoliths. AB - Ion-exchange porous adsorbent media having intermediate and low surface densities of dextran polymer grafted on the surface of the throughpores of polymeric monoliths are constructed and characterized by a molecular dynamics modeling and simulation approach that has also been shown to be effective in the construction and characterization of porous ion-exchange adsorbent media whose number of immobilized dextran polymer chains per unit surface area is high. The activation step that prepares the surface of the pores of the dextran polymer layer for the immobilization of the charged ligands insignificantly affected the pore structure of the dextran polymer layer, while this was found to not be the case for previously studied systems that involved high dextran polymer surface densities. Compared to the high dextran polymer density system studied previously, the intermediate dextran polymer density system can generate significantly larger pores but still possesses relatively high interconnection and mutual steric support between dextran chains to exhibit similar structural characteristics and responses to charged ligand immobilization, including dextran layer thickness, stability, monomer distribution, ligand-induced compact chain structures, dextran layer shrinkage, distributions of ligands and counterions, and local nonelectroneutrality. The low dextran polymer density system having relatively isolated dextran chains and insufficient mutual steric support can result in even larger pores than those obtained in the intermediate dextran polymer density system, but a significantly thinner porous dextran polymer layer and different dextran monomer distributions are obtained in the low dextran polymer density system. More importantly, the gradient of the local nonelectroneutrality after the immobilization of the charged ligands is significantly smaller in magnitude in the low dextran polymer density system than that obtained in the system having intermediate dextran polymer density, and, despite a lack of porous layer depth to accommodate adsorbate biomolecules in large amounts, it could still be useful for the effective transport and adsorption of very large biomolecules. Compared with the polymeric monoliths without a porous dextran polymer layer grafted on the surface of their throughpores, the intermediate and low dextran polymer density systems explored and studied in this work provide pore structures with desirable characteristics for the effective transport of adsorbate biomolecules and substantially larger effective surface areas and throughput capacities for the adsorption of the adsorbate biomolecules. PMID- 19555552 TI - Optimization of poly(GMA-co-EDMA) monolithic support for trypsin nanoreactor fabrication. AB - Fabrication of poly(glycidyl methacrylate-co-ethylene dimethacrylate) [also referred to as poly(GMA-co-EDMA)] monoliths was optimized as supporting material for trypsin digestion nanoreactors. Reaction parameters, such as polymerization time, porogen concentration, and monomer to crosslinker ratios, were evaluated in respect to the permeability of the resulting monolith and their effect on digestion efficiency, estimated by mass spectrometric analysis of a model protein cytochrome C. The structural homogeneity of the resulting monolithic support was checked by scanning electron microscopy. The best nanoreactor performance, measured by the reduction of nanoreactor backpressure and increased sequence coverage of cytochrome C, was achieved with 8% 2-octanol (porogen) 20%/20% glycidyl methacrylate to ethylene glycol dimethacrylate ratio and 5 h of polymerization time. Digestion of as low as 3 microg of cytochrome C with 77% sequence coverage was obtained using the optimized trypsin nanoreactor. PMID- 19555553 TI - Highly sensitive LC-MS-MS analysis of a pharmaceutical compound in human plasma using monolithic phase-based on-line extraction. AB - Monolithic columns have been used in recent years for fast chromatographic separation due to their high permeability and low backpressure. We have explored the potential of monolithic material as sample preparation tool in bioanalytical applications. By taking advantage of monolithic columns' online concentration capability, we have developed a highly sensitive liquid chromatography-tandem mass spectrometry assay for quantitative determination of a pharmaceutical compound in human plasma. The assay was fully validated to satisfy the requirements of precision and accuracy, selectivity, matrix effect, and reproducibility. A linear dynamic range from 0.011 ng/mL to 12.3 ng/mL was established as the calibration standard. The percentage of bias for quality control samples was between -9.9% and -2.5%. Coefficient of variation, a measurement of precision, was within 9.9%. On-line extraction with monolithic support provided adequate sample cleanup and on-line concentration of the analyte. The assay exhibited good tolerance to matrix effect and has been applied successfully to a clinical study. The incurred sample analysis showed that original and repeat values were within +/-10.1% for all assay samples. PMID- 19555554 TI - Development and validation of a rapid column-switching high-performance liquid chromatographic method for the determination of Lamotrigine in human serum. AB - This study describes a simple and sensitive column-switching high-performance liquid chromatographic method with UV detection for the determination of Lamotrigine in 50 microL of serum. After solid-phase extraction of Lamotrigine on an Oasis HLB extraction precolumn (20 x 3.9 mm; dp: 25 microm), chromatographic separation was achieved at 30 degrees C on a Chromolith RP-18e column (50 mm x 4.6 mm i.d.) using a solution of 20% acetonitrile in 15 mM phosphate buffer (pH 7.0) as the mobile phase, at a flow-rate of 2.0 mL/min. The eluant was detected at 215 nm. The retention time for Lamotrigine was 1.28 min. The total analysis time was ca. 5 min. However, the overlap of sample preparation, analysis, and reconditioning of the precolumn increased the overall sample throughput to one injection every 3 min. The method was validated for system suitability, linearity, precision, accuracy, robustness, and limit of quantitation. The linearity of the calibration lines, expressed by the linear correlation coefficient, was better than 0.9996. Recovery studies achieved from Lamotrigine spiked plasma samples showed values greater than 93%, demonstrating the excellent extraction efficiency of the precolumn. Intra- and inter-day precision were generally acceptable; the coefficient of variation was < 2.3% in all cases. The detection limits for Lamotrigine at a signal-to-noise ratio of 3 was 0.002 microg/mL when a sample volume of 50 microL was injected. However, it was possible to enhance the sensitivity further by injecting larger volumes, up to 200 microL. The method was shown to be robust and the results were within the acceptable range. The method was successfully applied to the determination of Lamotrigine in human serum samples of patients submitted to Lamotrigine therapy. PMID- 19555555 TI - Simultaneous determination of three antioxidants in foods and cosmetics by flow injection coupled to an ultra-short monolithic column. AB - The combination of an ultra-short C18 monolithic column (5 mm long) with flow injection analysis results in a versatile and efficient system that has been used for the determination of three antioxidants [propylgallate (PG), butylhydroxyanisole (BA), and butylhydroxytoluene (BT)]. Due to the wide variety of polarities of the analytes, two different carriers (carrier A: methanol-water 42% and carrier B: methanol-water 70%) were able to separate the analytes in only 85 s. The applicable concentration range, the detection, and the relative standard deviation (n=10) were: for PG, from 2.77 to 300 microg/mL, 0.84 microg/mL, 2.84%; for BA, between 1.51 and 300 microg/mL, 0.46 microg/mL, and 2.70%; and for BT, between 1.65 and 100 microg/mL, 0.55 microg/mL, and 2.22%, respectively. The method was applied and validated satisfactorily for the determination of PG, BA, and BT in food and cosmetic samples. PMID- 19555557 TI - [Current status and problems of interventional catheterization for pediatric ventricular septal defect in China]. PMID- 19555556 TI - Electrochromatographic evaluation of diol-bonded silica monolith capillary column for separation of basic compounds. AB - A diol-bonded silica monolith capillary column as polar stationary phase was successfully prepared for capillary electrochromatography. The preparation of monolithic stationary phase was based on the individual silica matrix forming and subsequent chemical bonding by (3-glycidoxypropyl) trimethoxysilane to produce the desired function. The diol-bonded silica monolith has been successfully employed in the electrochromatographic separation of alkaloids. The effects of experimental parameters, such as the volume fraction of the organic modifier, pH value, and ionic strength of the buffer on the retention behavior of the solutes were investigated. Column efficiencies greater than 110,000 plates/m for separation of basic compounds were obtained. It was observed that retention of alkaloids on the diol-bonded silica monolith was mainly contributed to a reversed phase and cation-exchange mechanism, and electrophoresis of basic compounds also played a role in separation. PMID- 19555558 TI - [To learn the correct clinical consideration and standard medical behaviors from the evolution of management of congenital heart disease]. PMID- 19555559 TI - [Expedite the construction of research and medical care system for child development and behavior in China]. PMID- 19555560 TI - [Results of surgical treatment of 1,387 infants under 6 months of age with congenital heart disease]. AB - OBJECTIVE: To summarize and review the result of surgical repair of congenital heart disease in infants under 6 months of age. METHODS: Between January 1997 and December 2007, 1387 infants under 6 months of age with congenital heart disease were operated on. There were 675 cases with ventricular septal defect with pulmonary hypertension (VSD/PH), 138 with complete transposition of the great arteries (TGA), 155 with tetralogy of Fallot (TOF), 111 with totally abnormal pulmonary venous connection (TAPVC), 54 with coarctation of aorta or interrupted aortic arch with ventricular septal defect [CoA(IAA)/VSD], 46 with double outlet right ventricle (DORV), 25 with pulmonary atresia with ventricular septal defect (PA/VSD), 24 with pulmonary atresia with intact interventricular septum (PA/IVS) and so on. The operative procedure was dependent on different diseases. Follow-up has been conducted in patients with some complex congenital heart diseases. RESULTS: In the recent 11 years, the number of surgical repair in infants under 6 months of age, including neonates, with congenital heart disease has been increased. In contrast to the early phase when ventricular septal defect was the major disease treated with surgery, infants with complex congenital heart disease account for half of all cases treated with surgery now. In the meantime, the surgical mortality has been decreased year after year. There were 110 deaths in our group and the total mortality was 7.9% (110/1387). With improvement of surgical procedure, the mortality was decreased from 11.5% - 14.4% in 1997 - 2003 to 8.6% - 8.9% in 2004 - 2005 to 3.3% - 3.8% in 2006 - 2007. Follow-up data were available for 98 patients in TGA (83.8%, 98/117), 79 in TAPVC (87.8%, 79/90), 68 in TOF (48.2%, 68/141), 13 in PA/VSD (65%, 13/20) and 19 in PAA/IVS (95%, 19/20). The duration of follow-up ranged from 3 to 86 months. There were 16 late deaths, 4 in TGA, 10 in TAPVC and 2 in PA/VSD patients. The majority were asymptomatic on follow-up. Mild residual obstruction was seen in 4 cases with TAPVC. Pulmonary hypertension was seen in 5 cases with TAPVC. Mild aortal valve regurgitation, pulmonary valve regurgitation and tricuspid valve regurgitation were seen in 23 cases with TGA (23.5%, 23/98). Some PA/VSD patients had second operation. CONCLUSIONS: Most symptomatic neonates and infants younger than 6 months with critical congenital heart defects can undergo corrective operation under acceptable risk. Due to improvements in perioperative, anaesthetic, surgical, and postoperative care, contemporary hospital mortality can be reduced to 3.3% - 3.8%. Palliative procedures still play an important role in the staged treatment of severe complex heart defects in neonates and infants younger than 6 months of age. PMID- 19555561 TI - [Therapeutic effect of Cheatham-Platinum stent implantation for vessel stenosis associated with congenital heart disease in children and adolescents]. AB - OBJECTIVE: Although NuMED Cheatham-Platinum (CP) stent was specifically designed to treat vascular obstructions associated with congenital heart disease (CHD), its application in pediatric patients is relatively uncommon, especially in the pulmonary artery stenosis. The aim of this study was to evaluate the immediate-, early- and intermediate-term results of CP stent implantation in the treatment of vessel stenosis associated with CHD in children and adolescents. METHODS: From August 2005 to May 2007, 5 consecutive patients (3 boys and 2 girls) diagnosed as vascular stenosis associated with CHD underwent CP stent implantation in our institution. One patient had native coarctation of the aorta (CoA) and four patients had pulmonary artery stenosis. The median age and weight of patients were 12 years (range 4 - 15 years) and 24 kg (range 20 - 51 kg), respectively. The CP stent and NuMED Balloon-in-Balloon catheter were selected according to digital subtracted angiography measurements. After checking for correct position by angiography, the inner balloon and outer balloon inflated successively to expand the stent to desired diameter. RESULTS: Totally 6 stent placement procedures were performed and 8 CP stents (8-zig, 22 - 39 mm in length) were implanted in these 5 patients. All stents but one in a case of right pulmonary artery stenosis were immediately successfully placed in the target lesions without displacement during the procedures. For this case, a repeat procedure was performed and a second CP stent was reimplanted successfully 11 months later. After the procedure, the systolic pressure gradient across the stenosis decreased from (43.43 +/- 25.61) mm Hg (1 mm Hg = 0.133 kPa) to (3.29 +/- 3.09) mm Hg (t = 4.320, P < 0.01) and the narrowest diameter of the stenotic vessels increased from (6.86 +/- 2.04) mm to (13.44 +/- 4.02) mm (t = -4.508, P < 0.01). The percentage of pulmonary artery flow to the ipsilateral lung increased from 11.0% and 13.0% to 47.5% and 52.2% after the procedure in 2 cases of unilateral pulmonary artery branch stenosis, respectively. The ratio of right ventricular to aortic systolic pressure decreased from 62.3% and 72.2% to 27.0% and 33.3% in 2 cases of bilateral branch pulmonary artery stenosis, respectively. Upper limb blood pressure of one case of native CoA dropped greatly from 206/133 mm Hg to 156/95 mm Hg. During a median follow-up of 20 months (range 13 - 34 months), the results have been stable without complications except 2 stents which developed intrastent restenosis 6 months after the procedure. CONCLUSION: Our experience suggests that the CP stent implantation is safe and feasible for the treatment of vessel stenosis associated with CHD in children and adolescents. The immediate-, early- and interim results are encouraging, but long-term results remain to be further evaluated and demand many more cases to be studied. PMID- 19555562 TI - [Effect of doxycycline on the development of pulmonary hypertension induced by four methods in rats]. AB - OBJECTIVE: Based on establishment of four rat models of experimental pulmonary hypertension (PH), the authors examined the inhibition of matrix metalloproteinases (MMPs) by doxycycline and its effect on the development of PH and associated pulmonary vascular remodeling. METHOD: Healthy male Sprague-Dawley rats (weight 350 g to 400 g) were randomly divided into nine groups: Normal control group (N), four model groups (H, M, P, PM) and their corresponding drug intervention groups (HD, MD, PD, PMD) in which doxycycline was given by gavage at a 20 mg/kg daily dosage. On day 28 (day 35 for PM and PMD models), the animals were catheterized to record mean pulmonary arterial pressure (mPAP) and then sacrificed. Fulton Index [RV/(LV + S)] was measured immediately. Morphometric parameters, including percent vascular wall thickness and muscularization of non muscularized peripheral pulmonary arterioles were determined microscopically. The activity of MMPs was measured by gelatin zymography in the lung tissue. RESULTS: (1) Rats in all model groups (H, M, P, PM) developed significant pulmonary arterial hypertension and right ventricular hypertrophy in comparison with their corresponding drug intervention groups (HD, MD, PD, PMD) and normal control group (N) (P < 0.01). For example, mPAP (mm Hg)(1 mm Hg = 0.133 kPa):N: 18.10 +/- 1.45, H: 27.20 +/- 1.55, HD: 23.90 +/- 2.13; Fulton Inedx(%):N: 23.41 +/- 1.84, H: 34.44 +/- 2.70, HD: 27.55 +/- 2.45. (2) The percent vascular wall thickness (WT%) and percentage of muscularization of non-muscular pulmonary arterioles were significantly increased in all model groups compared with drug intervention groups and normal group (P < 0.01). For example, WT%:N: 10.90 +/- 3.11, H:41.41 +/- 5.21, HD: 17.73 +/- 3.12; Muscularization(%):N: 13.83 +/- 3.72, H: 44.93 +/- 2.43, HD: 29.89 +/- 4.45. (3) The activity of MMPs was inhibited by doxycycline effectively as assessed by gelatin zymography (P < 0.01). For example, the activity of MMP2 (A x 10(3)):N: 1.43 +/- 0.24, H: 3.58 +/- 0.28, HD: 2.29 +/- 0.31. CONCLUSION: Doxycycline attenuated PH and associated pulmonary vascular remodeling in all rat PH models. The study suggests that high expression and enhanced activity of MMPs may play a brutial role in the development of PH. Such phenomenon seems to be common in a variety of PH models of different etiology. PMID- 19555563 TI - [Progresses in studies on pathogenesis of Kawasaki disease]. PMID- 19555564 TI - [Natural history and interventional therapy of tachyarrhythmia in children]. PMID- 19555565 TI - [Ten-year changes in pathogen, antimicrobial susceptibility and clinical feature of children with bacterial meningitis]. AB - OBJECTIVE: Despite progress in antibiotic therapy and intensive care, childhood bacterial meningitis (BM) remains a devastating disease. We conducted this study to investigate the changes in clinical characteristics, the etiologic agents and antimicrobial susceptibility of BM during the past 10 years in children under 14 years of age. METHODS: These 126 patients were divided into two groups according to their date of admission. Group 1 included 64 patients admitted from January 1998 to December 2002, and group 2 included 62 cases admitted from January 2003 to December 2007. All pediatric medical charts of them were reviewed. RESULTS: The predominant isolated bacteria from CSF were coagulase-negative staphylococcus (17/62, 27.4%) and Escherichia coli (9/62, 14.5%) in group 2. The resistance rate of staphylococcus against oxacillin (MRS) was 68.4% (13/19) in group 2, significantly higher than that of group 1 (16.7%, 2/12). Among 126 cases, 42 had seizure attack and 16 had consciousness disturbance, the proportions of them in group 2 (11/62, 17.7%; 4/62, 6.4%) were lower than those in group 1 (31/64, 48.4%; 12/64, 18.8%, P < 0.05). Cases in group 2 survived with complications [13/62 (21.0%)] and sequelae [11/62 (17.7%)] were lower than those in group 1 (24/64, 37.5%, 23/64, 35.9%, P < 0.05), but the rate of empirical therapy modification in group 2 (21/62, 33.9%) was higher than that in group 1 (7/64, 10.9%, P < 0.01). CONCLUSION: The predominant bacteria in children with BM are staphylococcus and Escherichia coli in recent years. The antibiotic resistance rate of bacteria has been higher year after year. The clinical patterns of pediatric BM have changed with a decrease in clinically serious cases, complications and sequelae, but an increase in modification of empirical therapy. PMID- 19555566 TI - [Enzymatic diagnosis of 47 cases with mucopolysaccharidosis]. AB - OBJECTIVE: With the emergence of enzyme replacement therapy and progress in bone marrow transplantation, treatment of mucopolysaccharidosis (MPS) is much more promising than ever. In order to benefit from these therapies, determination of the defective enzyme is the prerequisite for any individual patient. To make definite diagnosis for patients suspected of having MPS clinically, the authors established six lysosomal enzymatic assays for leucocytes, including alpha-L iduronidase, iduronate-2-sulfatase, N-acetylgalactosamine 6-sulfatase, beta galactosidase, arylsulfatase B, beta-glucuronidase, which are the corresponding enzymes of type I, type II, type IVA, type IVB, type VI, and type VII, respectively. METHOD: Seventy patients suspected of having MPS were enrolled from outpatient clinics of the Department of Pediatric Endocrinologic, Genetic and Metabolic Diseases in Xinhua Hospital. Their ages spanned from 10 months to 25 years with the average age 5.7 years. Of them 49 were male and 21 were female. Leukocytes were isolated with Dextran from peripheral blood of suspected patients. Activity of leukocyte alpha-L-iduronidase, iduronate-2-sulfatase, N acetylgalactosamine 6-sulfatase, beta-galactosidase, beta-glucuronidase were measured using their specific artificial fluorescent substrates, while arylsulfatase B were determined by colorimetric assay with dipotassium 2-hydroxy 5-nitrophenyl sulfate as the substrate. RESULT: Of the 70 clinically suspected cases totally 47 were confirmed of having mucopolysaccharidosis, of whom 7 cases were type I, 28 cases type II, 12 cases type IVA. These data show that type II is the predominant form of MPS in China, succeeded by MPS type IVA. It was also noted that type II has the most variable clinical manifestations and 8 out of 12 type IVA patients had the unique lax joints. CONCLUSION: The present study suggest that type II might be the predominant form of MPS cases in China, followed by type IVA and type I. PMID- 19555567 TI - [Growth standardized values and curves based on weight for length/height, body mass index for Chinese children under 7 years of age]. AB - OBJECTIVE: To construct the growth standardized data and curves based on weight for length/height, body mass index (BMI) for Chinese children under the age of 7 years. METHODS: The fourth national growth survey of children under 7 years of age in the nine cities of China was performed in 2005 and from this survey, data of the 69 622 urban healthy children were used to set up the database for weight for length/height (length was measured for children under 3 years of age) and BMI. Measurement data were ascribed to rigorous methods of data collection and standardized procedures across study sites. Models of weight for length/height and BMI were established by the LMS method, respectively; 0.7 cm was added to the height values (3 to 7 years), then weight for length/height were fitted by the single model, and after the model was fitted, the weight-for-length percentile curves in the length interval 65.7 to 125.7 cm were shifted back by 0.7 cm to derive the weight-for-height standards corresponding to the height range 65 cm to 125 cm. The BMI standards based on the length and height values were separately constructed by two sets of the LMS models. The height (3 to 7 years) data adding 0.7 cm was combined to the length (0 to 3 years) data, then the BMI-for-age standard based on length (0 to 3 years) was constructed. Analogously, BMI-for-age standard based on height was established. To test the goodness of fit of the model, chi(2) test for each sex and age was carried out. RESULTS: Standardized values and curves based on percentiles and z-score for 45 - 105 cm weight-for length, 65 - 125 cm weight for height, 0 to 3 years BMI-for-age based on length and 2 to 7 years BMI-for-age based on height were generated respectively. On the whole, the Chinese child growth charts was slightly higher than the new WHO child growth standards, and slightly lower than the 2000 CDC growth charts. CONCLUSION: Weight-for-length/height and BMI-for-age growth charts newly established were national representative growth standards, so it was recommended as the Chinese child growth standards used in the whole country for child growth monitoring and nutritional evaluation. PMID- 19555568 TI - [Clinical manifestations, laboratory test, treatment and outcome of neuropsychiatric involvement in pediatric systemic lupus erythematosus patients]. AB - OBJECTIVE: To investigate the clinical manifestations, laboratory test, treatment and outcome of neuropsychiatric (NP) involvement in pediatric systemic lupus erythematosus (SLE) patients. METHODS: Seventy-seven patients with NP syndromes of SLE (NPSLE) seen from 1987 to 2007 were retrospectively reviewed. The relationship between the relative factors and the relapse of NPSLE was analyzed with logistic regression model. RESULTS: NPSLE was found in 17.3% of the SLE patients and 75% of the NPSLE patients the NP involvements occurred in the first 2 years of the onset of SLE. The most frequent NP manifestations were headache (31.8%) and seizure disorder (29.1%). In the active phases, the levels of Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) scores of the 92.2% patients were higher than 15 and belonged to severe lupus. The patients were accompanied frequently with fever (88.3%) and rash (84.4%). The most frequently involved organs were kidney (76.6%) and blood system (67.5%). In the active phases, the ANA was positive (98.7%), the level of ESR increased (86.3%), the level of complement profile decreased (72.7%). The cerebrospinal fluid (CSF) study, the CT, the MRI and the EEG were abnormal (90.1%, 60.7%, 54.8%, 73.9%, respectively). All the patients received glucocorticoids and immunodepressant treatment in which 79.2% received IV high-dose methylprednisolone (MP), 51.9% received intrathecal (IT) methotrexate (MTX) and dexamethasone (DXM), 26.0% received IVIG, 2 patients received autologous peripheral blood stem cell transplantation. The mortality was 9.0%. The rate of relapse was 22.0% and in 75.0% of relapsed patients the relapse occurred within 24 months from the onset of NPSLE. The SLEDAI scores related to the relapse of the NPSLE (chi(2) = 3.987, P = 0.0459, OR = 1.172, 95% CI 1.003 and 1.370). CONCLUSION: SLEDAI scores were significantly helpful in predicting recurrence of NPSLE. PMID- 19555569 TI - [Effect of maternal iron status on infant's iron level: a prospective study]. AB - OBJECTIVE: To investigate the effect of maternal iron levels in second trimester of pregnancy on the infants' early iron status, explore the relationship between maternal and infant's iron status, and analyze the main factor influencing anemia of the infants. METHODS: Totally 100 couples of mother-baby were followed up from the second trimester of the mothers' pregnancy till the babies were 3 - 5 months old in the Department of Primary Child Care from March 2006 to February 2007. The maternal venous blood samples were collected in the second and the third trimesters and were analyzed for Hb at the same time. The infants' venous blood samples were collected at the end of the follow-up visit and were analyzed for Hb. The values of sTfR of the infants were compared with the maternal iron status diagnostic criteria. The infant's weight was measured during the visits by two professional staff members. The physical growth was assessed with CDC2000 reference. RESULTS: (1) Among 100 couples of mother-baby, none of the Hb values was lower than 90 g/L during the whole period of survey. The prevalence of ID in the babies at the ages of 3-5 months was 39%. Among them 36 cases (36/39, 92.31%) were born to the mother with ID during pregnancy; 26 couples of mother-infant had normal iron status during the following study. (2) The ID' prevalence in the infants born to the mothers with ID was higher than in the babies born to the healthy mothers (chi(2) = 11.567, P < 0.005). The ID' prevalence in the infants born to the mothers with IDA was higher than in the babies born to the mothers with LID (chi(2) = 7.356, P < 0.01). (3) The sTfR values of infants born to the mother with ID during pregnancy were increased significantly (P < 0.01); the iron nutritional status did not show any significant difference between babies whose mothers were IDA and LID (P > 0.05). (4) The SF value of maternal venous blood was negatively correlated with the sTfR value of infant (r = -0.7552, P < 0.01). CONCLUSIONS: (1) The anemia of the infants aged 3 - 5 months was correlated with the iron shortage during the fetal stage. (2) The mild iron deficiency in the pregnant women during the second trimester could decrease the iron storage of fetus and then also could affect the iron status of the early infancy and cause anemia. PMID- 19555570 TI - [Relationship between pharmacokinetics and efficacy and toxicity of daunorubicin in children with acute leukemia]. AB - OBJECTIVE: To study relationship between daunorubicin (DNR) pharmacokinetics and efficacy and toxicity in children with acute leukemia. METHODS: (1) The concentration of DNR in plasma of children with acute leukemia was determined by high performance liquid chromatography (HPLC)-fluorescence detection method. Plasma was sampled frequently from the start of the infusion till the end of 24 h. DNR pharmacokinetics was studied by determination of the concentrations. (2) Efficacy and toxicity were monitored in each period after chemotherapy. Laboratory studies included examination of bone marrow, white blood cell count, electrocardiogram, echocardiogram, myocardial enzymogram, the liver and kidney function. RESULTS: (1) DNR was eliminated from plasma in a two-compartment manner. The maximum concentration was seen 1 - 3 h after infusion. Cmax was 63.50 microg/L. Tmax was 1.45 h. The concentration decreased quickly to a low level of about 11.52 microg/L from the end of 2 hours infusion. There was a large inter individual difference in pharmacokinetic parameters of DNR. The difference of CL was 9-fold, AUC was 8-fold, Cmax was 5-fold. (2) CL of male patients [57.99 L/(h.m(2))] was significantly lower than that of female patients [93.71 L/(h.m(2))] (P < 0.05). Tmax of children older than 6 years was 1.1 h, and that of children younger than 6 years was 1.8 h (P < 0.05); Cmax of children older than 6 years was 90.77 microg/L, younger than 6 years was 57.44 microg/L (P < 0.05). CONCLUSION: (1) There is a large inter-individual difference in pharmacokinetic parameters of DNR in children. It may predict individual variety of efficacy and toxicity. Therapeutic drug monitoring is important. (2) Male patients and children older than 6 years had a higher bioavailability and lower metabolism, toxicity may easily occur in such children, therefore they may need lower dose. PMID- 19555571 TI - [Plasma 25-(OH)D level and vitamin D receptor expression in apolipoprotein E deficient mice]. PMID- 19555572 TI - [Single nucleotide polymorphism in asthma predisposing genes in children in Shanghai]. PMID- 19555573 TI - [Analysis on antibacterial activity of common antibiotics to Mycoplasma pneumoniae]. PMID- 19555574 TI - [Detection of human metapneumovirus and human bocavirus in children with bronchiolitis in east Guangdong area]. PMID- 19555575 TI - [Comorbidity of attention deficit hyperactivity disorder clinical subtypes: a sample from a psychiatric clinic]. PMID- 19555576 TI - [Real time 3-dimensional echocardiography in independently guiding closure of atrial septal defect in 28 pediatric cases]. PMID- 19555577 TI - [Human cytomegalovirus infection in pediatric patients with hematopoietic stem cell transplantation]. PMID- 19555578 TI - [Application of 64-slice spiral computed tomography in the diagnosis of pediatric patients with tracheobronchial diseases]. PMID- 19555579 TI - [Carvedilol therapy in pediatric patients with Takayasu's arteritis complicated with congestive heart failure: a case report]. PMID- 19555580 TI - [Lymphoma in a pediatric patient with ataxia-telangiectasia in a family of two cases]. PMID- 19555581 TI - [Diagnosis and treatment of invasive pulmonary candidiasis: application of the guideline for diagnosis and treatment of invasive pulmonary fungal infection in children]. PMID- 19555582 TI - NCCN clinical practice guidelines in oncology: testicular cancer. PMID- 19555584 TI - NCCN clinical practice guidelines in oncology: kidney cancer. PMID- 19555586 TI - New treatments for renal cell carcinoma: targeted therapies. AB - Systemic treatment options for advanced renal cell carcinoma (RCC) have expanded considerably with the development of targeted therapies. Clear cell RCC commonly features mutation or inactivation of the von Hippel-Lindau gene and resultant overexpression of vascular endothelial growth factor (VEGF). The first drug to validate VEGF as a target in the treatment of clear cell RCC was the monoclonal antibody bevacizumab. Since then, anti-VEGF receptor therapy with multitargeted kinase inhibitors also has shown substantial efficacy. Sunitinib is now a standard first-line therapy for advanced disease and sorafenib is among the second-line treatment options. Other kinase inhibitors are in development. Mammalian target of rapamycin (mTOR) is a second validated therapeutic target as the mTOR inhibitor temsirolimus has been shown to prolong survival in first-line treatment of poor prognosis RCC of all histologies. Everolimus is an oral mTOR inhibitor and has been shown to prolong progression-free survival when used in second-line treatment. Non-clear cell and sarcomatoid RCC are both underrepresented in completed trials but are the subject of active research. Ongoing and planned studies will also evaluate the use of combinations of targeted agents, a strategy that is not advisable outside of clinical trials. Finally, postnephrectomy adjuvant treatment with targeted agents is not yet standard but is under investigation in phase III trials. PMID- 19555585 TI - Optimal management of localized renal cell carcinoma: surgery, ablation, or active surveillance. AB - Radical nephrectomy is historically accepted as standard treatment for localized renal cell carcinoma (RCC). However, the presentation of RCC has changed dramatically over the past 3 decades. Newer alternative interventions aim to reduce the negative impact of open radical nephrectomy, with the natural history of RCC now better understood. This article discusses current surgical and management options for localized kidney cancer. PMID- 19555587 TI - Non-clear cell renal cancer: features and medical management. AB - The treatment of metastatic renal cell carcinoma (RCC) has changed dramatically with the introduction of targeted therapies against vascular endothelial growth factor and the mammalian target of rapamycin. Because patients with clear cell histology account for more than 80% of patients with RCC, little evidence is available on treating patients with non-clear cell histologies. Most clinical trials have excluded them from enrollment, except for a randomized study investigating temsirolimus. Many retrospective studies on the use of sunitinib, sorafenib, and temsirolimus in patients with non-clear cell histology have shown response rates ranging from 3.7% to 16%. Prospective studies in non-clear cell histologies are ongoing. Although response rates may not be as high as those in patients with clear cell histologies, targeted therapy may provide a clinically meaningful response. New investigational therapies are on the horizon for papillary RCC--the most-common non-clear cell RCC histology--targeting pathways specific to this histology, such as the c-MET pathway. PMID- 19555588 TI - NCCN task force: clinical utility of PET in a variety of tumor types. AB - Use of PET is widespread and increasing in the United States, mainly for oncologic applications. In November 2006, the National Comprehensive Cancer Network (NCCN) gathered a panel of experts to review the literature and develop clinical recommendations for using PET scans in lymphoma and non-small cell lung, breast, and colorectal cancers. However, because its use is not restricted to these diseases, and evidence is accumulating for its application in other types of cancers, NCCN convened a second meeting in December 2008 to expand on the initial report. A multidisciplinary panel met to discuss the current data on PET application for various tumor types, including genitourinary, gynecologic, pancreatic, hepatobiliary, thyroid, brain, small cell lung, gastric, and esophageal cancers, and sarcoma and myeloma. This report summarizes the proceedings of this meeting, including discussions of the background of PET, the role of PET in oncology, principles of PET use, emerging applications, and possible future developments. PMID- 19555590 TI - Psychological interventions for postnatal depression: cluster randomised trial and economic evaluation. The PoNDER trial. AB - OBJECTIVES: To investigate outcomes for postnatal women attributed to special training for health visitors (HVs) in systematically identifying postnatal depression and delivering psychologically informed interventions, and to establish the cost-effectiveness of the intervention. DESIGN: A pragmatic randomised cluster trial with clusters allocated to experimental HV training arms or control, with an 18-month follow-up. SETTING: GP practices in the former Trent Regional Health Authority. PARTICIPANTS: Women registered with participating GP practices who became 36 weeks pregnant during the recruitment phase of the trial, had a live baby and were on a collaborating HV's caseload for 4 months postnatally. INTERVENTION: HV training in the assessment of postnatal women, combined with either cognitive behavioural approach (CBA) or person-centred approach (PCA) sessions for eligible women, plus the option of a selective serotonin reuptake inhibitor if indicated. MAIN OUTCOME MEASURES: The primary outcome was the proportion of at-risk women with a 6-month Edinburgh Postnatal Depression Scale (EPDS) score > or = 12. The primary comparison was between at risk women in the combined clusters randomised to HV training and women in practices randomised to provide HV usual care. The secondary comparison was to determine any differences between the proportions of women with a 6-month EPDS score > or = 12 in the CBA and PCA groups. RESULTS: HVs in 101 clusters in 29 primary care trusts collaborated in the study. From 7649 eligible women 4084 (53.4%) consented to take part: 17.3% (595/3449) of women who returned a 6-week questionnaire had a 6-week EPDS score > or = 12 and were at-risk women; 70.3% (418/595) of at-risk women had a 6-month EPDS score available. In total, 45.6% (67/147) of control group (CG) at-risk women had a 6-month EPDS score > or = 12 versus 33.9% (93/271) of intervention group (IG) women (p = 0.036). A total of 32.9% (46/140) of at-risk women in the CBA group versus 35.1% (46/131) in the PCA group had a 6-month EPDS score > or = 12 (p = 0.74). The CG mean 6-month EPDS score for at-risk women was 11.3 (SD 5.8) versus 9.2 (SD 5.4) for the IG (p = 0.002) and this remained statistically significant after adjusting for 6-week variables (p = 0.001). In total, 16.4% (150/914) of all women in the CG had a 6 month EPDS score > or = 12 compared with 11.7% (205/1745) in the IG (p = 0.003). The CG mean 6-month EPDS score for all women was 6.4 (SD 5.2) compared with 5.5 (SD 4.7) for the IG (p < 0.001). The economic analysis results showed a consistent pattern of psychological approaches being cost-effective at funding levels used by the National Institute for Health and Clinical Excellence. CONCLUSIONS: HV training was effective compared with HV usual care in reducing the proportion of at-risk women with a 6-month EPDS score > or = 12, with a wide confidence interval for the estimated intervention effect, suggesting that the true treatment effect may be small. The effect remained for 1 year. The economic evaluation demonstrated that the HV intervention was highly likely to be cost effective compared with the control. There was no difference in outcomes between the CBA and the PCA groups. PMID- 19555592 TI - Now available: German annual epidemiological report on notifiable diseases 2008. PMID- 19555593 TI - Short summary of Swedres 2008, a report on antimicrobial utilisation and resistance in humans in Sweden. PMID- 19555594 TI - Clinical and epidemiological aspects of parvovirus B19 infections in Ireland, January 1996-June 2008. AB - Parvovirus B19 infection may be mistakenly reported as measles or rubella if laboratory testing is not performed. As Europe is seeking to eliminate measles, an accurate diagnosis of fever/rash illnesses is needed. The main purpose of this study was to describe the epidemiological pattern of parvovirus B19, a common cause of rash, in Ireland between January 1996 and June 2008, using times series analysis of laboratory diagnostic data from the National Virus Reference Laboratory. Most diagnostic tests for presumptive parvovirus B19 infection were done in children under the age of five years and in women of child-bearing age (between 20-39 years-old). As a consequence, most of the acute diagnoses of B19 infection were made in these populations. The most commonly reported reasons for testing were: clinical presentation with rash, acute arthritis, influenza-like symptoms or pregnancy. The time series analysis identified seasonal trends in parvovirus B19 infection, with annual cycles peaking in late winter/spring and a six-year cycle for parvovirus B19 outbreaks in Ireland. PMID- 19555595 TI - Trends in the epidemiology of dengue fever and their relevance for importation to Europe. AB - Dengue fever continues to spread globally, causing major epidemics and putting major strain on health systems in affected countries. For imported dengue in Europe, south east Asia is the most important region of origin, followed by Latin America, the Indian subcontinent, the Caribbean, and Africa. Information regarding mosquito protective measures is highly recommended for all travellers to affected areas. PMID- 19555589 TI - NCCN Task Force Report: Bone Health in Cancer Care. AB - Bone health and maintenance of bone integrity are important components of comprehensive cancer care in both early and late stages of disease. Risk factors for osteoporosis are increased in patients with cancer, including women with chemotherapy-induced ovarian failure, those treated with aromatase inhibitors for breast cancer, men receiving androgen-deprivation therapy for prostate cancer, and patients undergoing glucocorticoid therapy. The skeleton is a common site of metastatic cancer recurrence, and skeletal-related events are the cause of significant morbidity. The National Comprehensive Cancer Network (NCCN) convened a multidisciplinary task force on Bone Health in Cancer Care to discuss the progress made in identifying effective screening and therapeutic options for management of treatment-related bone loss; understanding the factors that result in bone metastases; managing skeletal metastases; and evolving strategies to reduce bone recurrences. This report summarizes presentations made at the meeting. PMID- 19555596 TI - Chikungunya infection confirmed in a Belgian traveller returning from Phuket (Thailand). AB - Chikungunya infection has been increasingly reported in international travellers following its epidemic re-emergence in the Indian Ocean islands in 2006 and its spread to southern Asia thereafter. We describe the first case of chikungunya in a Belgian traveller returning from Phuket, Thailand and discuss the potential implications of chikungunya cases imported to European countries for patient management and public health. PMID- 19555597 TI - Invitation to become part of the European Travel Medicine Inventory. PMID- 19555598 TI - A variety of respiratory viruses found in symptomatic travellers returning from countries with ongoing spread of the new influenza A(H1N1)v virus strain. AB - Clinical specimens from 79 symptomatic individuals with a recent history of travel to countries with verified transmission of influenza A(H1N1)v (North America) were tested with a multiple real-time PCR targeting a broad range of agents that may cause acute respiratory infection. This analysis revealed that besides four cases of influenza A(H1N1)v, other respiratory viruses were diagnosed in almost 60% of the samples. These observations are a reminder that many different viral transmissions occur simultaneously in countries with ongoing spread of influenza A(H1N1)v. The findings demonstrate that the definition of suspected cases by clinical and epidemiological criteria has only a poor capacity for discriminating influenza A(H1N1)v from other viral infections. PMID- 19555599 TI - School closure is currently the main strategy to mitigate influenza A(H1N1)v: a modeling study. AB - Mathematical modeling was employed on recent epidemiological data from Mexico in order to assess the impact of intervention strategies on the spread of influenza A(H1N1)v in the setting of the European region. Active surveillance that will ensure timely treatment and home isolation of symptomatic cases in combination with school closure seem to form an efficient strategy to control the spread of influenza A(H1N1)v. PMID- 19555600 TI - Epidemiology of influenza A(H1N1)v virus infection in Japan, May-June 2009. AB - Between 9 May and 4 June 2009, a total of 401 laboratory-confirmed cases of influenza A(H1N1)v virus were reported in Japan, from 16 of the 47 Japanese prefectures. The two areas most affected were Osaka prefecture and Kobe city where outbreaks in high schools occurred leading to school closures. To date all cases have had symptoms consistent with seasonal influenza and no severe or fatal cases have been reported. PMID- 19555601 TI - Virological surveillance of human cases of influenza A(H1N1)v virus in Italy: preliminary results. AB - In this report we describe the findings of laboratory-based surveillance of human cases of influenza A(H1N1)v virus infection in Italy, following the recent worldwide detection of this new virus among human population and the decision of the World Health Organization (WHO) to raise the level of pandemic alert. PMID- 19555602 TI - Imported fresh sugar peas as suspected source of an outbreak of Shigella sonnei in Denmark, April-May 2009. AB - We report on an outbreak of Shigella sonnei infections involving ten cases notified through the laboratory surveillance system in Denmark in April and May. The likely source was consumption of fresh, raw sugar peas (sugar snaps) imported from Africa. This conclusion was based on interviews with cases and on the occurrence of a similar outbreak one month later in Norway. Fresh imported produce may occasionally be contaminated with pathogenic bacteria even when sold as ready-to-eat. PMID- 19555603 TI - Shigella sonnei infections in Norway associated with sugar peas, May-June 2009. AB - In May 2009, the Norwegian Institute of Public Health (NIPH) identified a possible outbreak of Shigella sonnei infection involving four cases. Additionally, five suspected cases in two separate households were reported. Inspectors from the Norwegian Food Safety Authority (NFSA) visited the two households and found an unopened package of sugar peas imported from Kenya in one of the households. One sample from the sugar peas was positive for Shigella sonnei by two PCR methods. Based on this result and information from patient interviews, the NFSA prohibited all sales of sugar peas imported from Kenya. PMID- 19555604 TI - Influenza A(H1N1)v in the southern hemisphere--lessons to learn for Europe? PMID- 19555605 TI - Evaluation of lung ultrasound for the diagnosis of pneumonia in the ED. AB - OBJECTIVES: The aim of this study is to assess the ability of bedside lung ultrasound (US) to confirm clinical suspicion of pneumonia and the feasibility of its integration in common emergency department (ED) clinical practice. METHODS: In this study we performed lung US in adult patients admitted in our ED with a suspected pneumonia. Subsequently, a chest radiograph (CXR) was carried out for each patient. A thoracic computed tomographic (CT) scan was made in patients with a positive lung US and a negative CXR. In patients with confirmed pneumonia, we performed a follow-up after 10 days to evaluate clinical conditions after antibiotic therapy. RESULTS: We studied 49 patients: pneumonia was confirmed in 32 cases (65.3%). In this group we had 31 (96.9%) positive lung US and 24 (75%) positive CXR. In 8 (25%) cases, lung US was positive with a negative CXR. In this group, CT scan always confirmed the US results. In one case, US was negative and CXR positive. Follow-up turned out to be always consistent with the diagnosis. CONCLUSION: Considering that lung US is a bedside, reliable, rapid, and noninvasive technique, these results suggest it could have a significant role in the diagnostic workup of pneumonia in the ED, even if no sensitivity nor specificity can be inferred from this study because the real gold standard is CT, which could not be performed in all patients. PMID- 19555606 TI - The effect of prolonged ED stay on outcome in patients with necrotizing fasciitis. AB - OBJECTIVE: Overcrowding in hospitals, especially in EDs, is a serious problem in the United States, Europe, and Taiwan. However, the association between prolonged ED boarding stay and mortality in patients with necrotizing fasciitis remains underinvestigated. METHODS: This was a retrospective study. A total of 195 patients were enrolled and analyzed. The sample was divided into 2 groups: nonmortality and mortality. A stepwise logistic regression model was developed to investigate 3 factors of clinical relevance predicting patient mortality. RESULT: The results of the stepwise logistic regression analysis revealed that hypotension (odds ratio [OR], 32.9; 95% confidence interval [CI], 6.9-156.0) and prolonged ED boarding stay (OR, 3.4; 95% CI 1.3-8.6) were both associated with higher mortality. Early operation (OR: 0.16; 95% CI: 0.06-0.45) was associated with lower mortality. CONCLUSION: Prolonged ED boarding stay was associated with increased mortality in patients with necrotizing fasciitis. Early operation (within 24 hours of ED arrival) was associated with decreased mortality. PMID- 19555607 TI - Air pollution and daily ED visits for migraine and headache in Edmonton, Canada. AB - BACKGROUND: A variety of environmental factors have been identified as possible triggers for migraine and other headache syndromes. OBJECTIVE: We analyzed associations between air pollution and emergency department (ED) visits for migraine and headache. METHODS: Analysis was based on 56,241 ED visits for migraine and 48,022 ED visits for headache to Edmonton hospitals between 1992 and 2002. A Poisson model of counts hierarchically clustered by day of week, month, and year was applied using generalized linear mixed models. Temperature and relative humidity were included as covariates. RESULTS: Females accounted for 78.5% of migraine visits and 56.3% of headache visits. An interquartile range (IQR) increase (6.2 microg/m3) in daily average particulate matter of median aerodynamic diameter less than 2.5 microm (PM2.5) was associated with increases in visits of 3.3% for migraine (95% confidence interval [CI]: 0.6-6.0), lagged 2 days, and 3.4% for headache (95% CI: 0.3-6.6), lagged 0 days, among females in the cold season (October-March). PM2.5 was also associated with cold season migraine visits among females at lag 0 and 1 day (P < .1). In the warm period (April-September), a 2.3-ppb IQR increase in sulfur dioxide was associated with a 2.5% increase in migraine visits (95% CI: 0.3-4.6) among females, whereas a 12.8 ppb IQR increment in nitrogen dioxide was associated with a 6.8% increase in headache visits (95% CI: 1.5-12.5) for males. CONCLUSIONS: Findings provide preliminary evidence of an association between air pollution and ED visits for migraine and nonspecific headache. Findings were most consistent for particulate matter. PMID- 19555608 TI - Bedside prediction of increased filling pressure using acoustic electrocardiography. AB - BACKGROUND: Patients presenting with acute dyspnea are often a diagnostic dilemma. A bedside tool that accurately and rapidly identifies increased left ventricular (LV) filling pressure would be helpful. We evaluated acoustic electrocardiography for this purpose. METHODS: We pooled 3 cohorts of patients for this analysis. Inclusion criteria required acoustic electrocardiography and echocardiography within 4 hours of each other. Increased LV filling pressure was defined as a pseudonormal or restrictive filling pattern on echocardiography. Area under the receiver operating characteristic curve (AUC) assessed multivariable model accuracy. RESULTS: The median age of the 324 patients was 61 years (range, 19-90 years), 67% were male, and 82% had a history of heart failure. The final multivariable model included mean LV systolic time, S(3) score, maximum negative area of the P wave, and the QTc interval. The AUC was 0.83 (95% confidence interval, 0.78-0.88). Although B-type natriuretic peptide (BNP) was an independent predictor of estimated increased filling pressure when considered alone (odds ratio = 1.002, 95% confidence interval, 1.000-1.003, P = .002), when added to the acoustic model, it did not improve overall model accuracy. In the subset of patients with indeterminate BNP levels (100-500 pg/mL), the acoustic model was more accurate than BNP (AUC = 0.82 vs 0.71). CONCLUSIONS: Bedside acoustic electrocardiography predicted echocardiographic correlates of increased pressures with high accuracy. For patients with an indeterminate BNP level (100-500 pg/mL), the acoustic electrocardiography model was superior to BNP. Prospective model validation is warranted. PMID- 19555609 TI - The importance of urine 5-hydroxyindoleacetic acid levels in the early diagnosis of acute appendicitis. AB - PURPOSE: Acute appendicitis is one of the most common surgical emergencies. Diagnosis is usually made depending on the presenting history, clinical evaluation, and laboratory tests. The aim of this study was to investigate the role of urinary 5-hydroxyindoleacetic acid (U-5-HIAA) in the early diagnosis of acute appendicitis. METHODS: Thirty-five pigmented male rabbits were divided into 5 groups. Group 1 is the control (n = 7); group 2 is the sham (n = 10). The appendix was ligated from its base, and an appendectomy was performed after 12, 24, 36 hours in group 3 (n = 7), group 4 (n = 7), and group 5 (n = 7), respectively. Spot urine samples were obtained for U-5-HIAA determination, and appendectomy tissues were examined histopathologically. RESULTS: Acute appendicitis was diagnosed in all animals in group 3, group 4, and group 5, and the mean levels of U-5-HIAA in group 3 were higher than in the other groups. The mean of U-5-HIAA levels between animals with appendicitis and those without showed a significant difference (P = .003). The U-5-HIAA cutoff point of 4.15 mg/g creatinine had a sensitivity of 85%, a specificity of 64.29%, and an accuracy of 76% (area under curve = 0.805) for acute appendicitis. The probability of acute appendicitis is found to be 10, 2 times more when the U-5 HIAA level is greater than 4.15 mg/g creatinine. CONCLUSION: We have concluded that spot U-5-HIAA level increases significantly in the early stages of acute appendicitis. PMID- 19555610 TI - Prolonged TASER use on exhausted humans does not worsen markers of acidosis. AB - OBJECTIVE: There are safety concerns about TASER conducted electrical weapon (CEW) use on humans, and there have been media reports of adverse human outcomes after CEW exposure. Conducted electrical weapons are often used on physically exhausted subjects. A single CEW application of a CEW is generally accepted to be 5 seconds of exposure. Some exposures in reality involve more than 5 seconds. We sought to determine if a prolonged (15 seconds) CEW exposure on exhausted humans caused acidosis, hyperkalemia, serum lactate change, or troponin change. METHODS: This was a prospective study of generally healthy human volunteers. Medical histories and baseline serum values were obtained, and several of the volunteers did have acute or chronic medical problems. Subjects underwent an exercise protocol until subjective exhaustion. Exhaustion was defined by the volunteer no longer being able to perform the exercise at a given pace. Blood was drawn immediately (defined as within 20 seconds) after exercise and was immediately followed by a 15-second CEW exposure. Blood was drawn immediately after exposure and again at 16 to 24 hours after exposure. Blood was analyzed for pH, pco(2), potassium, lactate, and troponin. Data were compared using Wilcoxon signed rank tests. RESULTS: There were 38 subjects enrolled with an average age of 39 years. The following health conditions were reported among the volunteers: hypertension (2), gastritis/reflux (2), active respiratory tract infections (3), asthma (2), chronic muscular pain conditions (4), pituitary adenoma (1) and glaucoma (1). Sixteen volunteers reported use of prescription medication at the time of their participation. The median initial pH of 7.38 (interquartile range [IQR], 7.35 7.40) decreased to 7.23 (IQR, 7.19-7.31) immediately after exercise. Immediately after exposure, median pH was 7.22 (IQR, 7.18-7.25). It was 7.39 (IQR, 7.37-7.43) at 24 hours. The pCO2 increased from 46.3 (IQR, 43.0-54.5) to 57.4 (IQR, 49.9 67.7) immediately after exercise, decreased to 51.3 (IQR, 44.4-65.0) immediately after exposure, and was 46.3 (IQR, 42.7-51.7) at 24 hours. Lactate increased from a median of 1.65 (IQR, 1.14-2.55) to 8.39 (IQR, 6.98-11.66) immediately after exercise, increased to 9.85 (IQR, 7.70-12.87) immediately after exposure, and was 1.02 (IQR, 0.91-1.57) at 24 hours. Serum potassium increased from 3.9 (IQR, 3.8 4.4) to 4.2 (IQR, 4.0-4.9) immediately after exercise, decreased to 3.8 (IQR, 3.7 4.4) immediately after exposure, and was 4.1 (IQR, 3.9-4.6) at 24 hours. No troponin elevations were detected. CONCLUSION: Prolonged CEW application on exhausted humans was not associated with worsening change in pH or troponin. Decreases in pCO2 and potassium and a small increase in lactate were found. Worsening acidosis theories due to CEW use in this population are not supported by these data. PMID- 19555611 TI - A test of syndromic surveillance using a severe acute respiratory syndrome model. AB - OBJECTIVES: We describe a field simulation that was conducted using volunteers to assess the ability of 3 hospitals in a network to manage a large influx of patients with a potentially communicable disease. This drill provided the opportunity to evaluate the ability of the New York City Department of Health and Mental Hygiene's (NYC-DOHMH) emergency department chief complaint syndromic surveillance system to detect a cluster of patients with febrile respiratory illness. METHODS: The evaluation was a prospective simulation. The clinical picture was modeled on severe acute respiratory syndrome symptoms. Forty-four volunteers participated in the drill as mock patients. RESULTS: Records from 42 patients (95%) were successfully transmitted to the NYC-DOHMH. The electronic chief complaint for 24 (57%) of these patients indicated febrile or respiratory illness. The drill did not generate a statistical signal in the NYC-DOHMH SaTScan analysis. The 42 drill patients were classified in 8 hierarchical categories based on chief complaints: sepsis (2), cold (3), diarrhea (2), respiratory (20), fever/flu (4), vomit (3), and other (8). The number of respiratory visits, while elevated on the day of the drill, did not appear particularly unusual when compared with the 14-day baseline period used for spatial analyses. CONCLUSIONS: This drill with a cluster of patients with febrile respiratory illness failed to trigger a signal from the NYC-DOHMH emergency department chief complaint syndromic surveillance system. This highlighted several limitations and challenges to syndromic surveillance monitoring. PMID- 19555612 TI - Does mad honey poisoning require hospital admission? AB - BACKGROUND: The aim of this study was to describe current patterns of monitoring and treatment of mad honey intoxication to make recommendations for a more standardized approach to care of patients with mad honey poisoning. METHODS: Patients presenting to emergency departments because of honey poisoning between January and October 2007. Age, length of stay in the emergency department, pulse rate, and systolic and diastolic blood pressure are cited as mean +/- SD. RESULTS: Forty-seven cases presenting to the 3 health institutions during 2007 were investigated. It was determined that patients had ingested "mad" honey between 0.5 and 9 hours (mean +/- SD, 2.8 +/- 1.8 hours) before presentation. Patients' pulse rates were 30 to 77/min (mean +/- SD, 46.6 +/- 12.1/min), and systolic blood pressure ranged from 50 to 140 mm Hg (mean +/- SD, 46.6 +/- 12.1 mm Hg). Patient rhythms on arrival were determined as 37 (7.7%) sinus bradycardia, 6 (12.8%) nodal rhythm, 3 (6.4%) normal sinus rhythm, and 1 (2.1%) complete atrioventricular block. Lengths of stay in hospital were 3.6 +/- 2.2 hours in the first university hospital, 22.2 +/- 3.8 hours in the second university hospital, and 3.4 +/- 1.7 hours in the state hospital. A 0.5 to 2 mg of atropine was given to all patients. CONCLUSIONS: Our study did not reveal any difference in complications or mortality between patients cared for with brief emergency department observation when compared with patients cared for with 1 day inpatient observation. PMID- 19555613 TI - Prevalence of incidental findings in trauma patients detected by computed tomography imaging. AB - STUDY OBJECTIVE: The primary intention of spiral computed tomography (SCT) in trauma patients is to identify significant injuries. However, unanticipated information is often discovered. We hypothesize that SCT often identifies clinically significant incidental findings in trauma patients. METHODS: This was a retrospective protocol chart review of consecutive adult trauma patients seen at a level I trauma center. A complete SCT was defined as computed tomography imaging of the head, cervical spine, chest, abdomen, and pelvis, thoracic, and lumbar spine. Incidental findings were classified into 2 categories: type 1, which requires urgent evaluation, and type 2, which requires informing the patient but does not mandate urgent follow-up. RESULTS: We reviewed 3246 patient charts and 3092 met inclusion criteria. Type 1 findings were reported in 990 (32.0%; 95% confidence interval [CI], 30.4%-33.7%) patients. Type 2 findings were found in 1274 (41.2%; 95% CI, 39.5%-42.9%) patients. Female sex (odds ratio, 1.38; 95% CI, 1.16-1.65) and older age (odds ratio, 2.61; 95% CI, 2.33-2.93) were independently associated with a higher prevalence of type 1 findings. There were 631 incidental findings concerning for neoplasm, which included 196 pulmonary nodules, 99 liver, 36 renal, 23 brain, and 11 breast masses. CONCLUSIONS: A significant number of trauma patients evaluated with SCT are diagnosed with potentially serious incidental findings. For long-term care and medicolegal concerns, physicians need to inform patients of these incidental findings and the need for further evaluation. PMID- 19555614 TI - Comparison of initial antibiotic choice and treatment of cellulitis in the pre- and post-community-acquired methicillin-resistant Staphylococcus aureus eras. AB - Treatment success of cellulitis in the age of community-acquired methicillin resistant Staphylococcus aureus (CAMRSA). A comparison of initial antibiotic choice and treatment success in the pre- and post-CAMRSA eras. OBJECTIVE: The objective of this study is to determine antibiotic prescribing patterns and treatment failure rates for cellulitis in the pre- and post-CAMRSA eras. METHODS: We performed an electronic chart review of patients seen in our emergency department with cellulitis in 2000 and 2005. Inclusion criteria included age 18 years or more and received a single oral antibiotic for cellulitis. Exclusion criteria were incision and drainage, surgery, or admission on initial visit. Treatment failure was defined as a repeat visit in the subsequent 30 days and a change in antibiotics, admission to the hospital, incision and drainage of abscess, or surgical intervention. Antibiotic-prescribing practices and treatment failure rates were then compared in the pre- and post-CAMRSA eras. RESULTS: There was a significant decrease in beta-lactam antibiotics and an increase in CAMRSA effective antibiotics prescribed in 2005 vs 2000. The difference in treatment failure rates of the beta-lactams and CAMRSA antibiotics was statistically insignificant. There has not been an increase in failure rates of the beta-lactam antibiotics for simple cellulitis since the emergence of CAMRSA. CONCLUSION: Our study demonstrates that prescribing practices for simple cellulitis have changed since the emergence of CAMRSA. This may not be appropriate because beta-lactam antibiotics perform as well as 'CAMRSA antibiotics' in our study. PMID- 19555615 TI - Prearrest signs of shock and respiratory insufficiency in out-of-hospital cardiac arrests witnessed by crew of the emergency medical service. AB - AIM: The objective of this study is to determine whether prearrest shock and respiratory insufficiency influence outcome in patients with emergency medical service-witnessed out-of-hospital cardiac arrest. METHODS: Analysis of data from a cardiac arrest database and data from the ambulance charts was performed. For the purpose of the study, shock was defined as prearrest heart rate below 40 or above 140/min, systolic blood pressure as below 90 mm Hg, and respiratory insufficiency as respiratory rate above 36 or oxygen saturation below 90%. Adjusted odds ratios (ORs) with 95% confidence intervals (CIs) were calculated. RESULTS: Of a total of 303 patients, 81% had prearrest shock or respiratory insufficiency. Mortality was higher in these patients indicated by fewer with return of spontaneous circulation (43% vs 75%, P < .001), and lower survival to hospital admission (31% vs 71%, P < .001) and to discharge (13% vs 59%, P < .001). Independent predictors of mortality were age (OR, 1.04; CI, 1.0-1.06), initial rhythm other than ventricular fibrillation or ventricular tachycardia (OR, 32.9; CI, 10.9-99.0), and respiratory insufficiency (OR, 4.2; CI, 1.4-12.5). CONCLUSIONS: Shock and respiratory depression are common among patients with out of-hospital cardiac arrest witnessed by the emergency medical service, and these patients have a high mortality when compared with patients without shock or respiratory failure. PMID- 19555616 TI - The correlation between elevated cardiac troponin I and pulmonary artery obstruction index in ED patients with pulmonary embolism. AB - OBJECTIVES: The aim of this study is to investigate the correlation between cardiac troponin I (cTnI) values and the pulmonary artery obstruction index assessed with spiral computed tomography (CT) scan in emergency department (ED) patients with pulmonary embolism (PE). METHODS: This is a retrospective cohort study of all 179 ED patients diagnosed with PE between December 2004 and January 2007. Study population consisted of 104 (58.1%) of 179 patients with PE in whom both cTnI was measured and a contemporaneously performed CT scan was available for review. In these patients, the levels of cTnI measured in the ED were correlated with the degree of pulmonary vascular obstruction determined by applying the modified Computed Tomography Obstruction Index to the spiral CT scan performed in the ED. RESULTS: Troponin values were elevated in 20 (19.2%) of 104 patients (95% confidence interval [CI], 11.6-26.8) with a mean cTnI concentration of 0.38 +/- 0.44 microg/L. Elevated cTnI value had a significant correlation with main pulmonary arteries involvement using the modified Computed Tomography Obstruction Index score (P = .0001). Elevated ED cTnI value had 53.8% (95% CI, 37.6-66) sensitivity and 92.3% (95% CI, 87-96.4) specificity, 70% (95% CI, 49-86) PPV, and 85.7% (95% CI, 80.7-90) NPV for predicting main pulmonary artery obstruction on CT. Increased cTnI values were highly correlated to intensive care unit admission of patients with PE (RR, 12.83; 95% CI, 3.87-42.4). CONCLUSIONS: Measuring cTnI value might be considered in ED patients who are suspected of having PE. Elevated cTnI should raise the clinician's concern for the possibility of central pulmonary vascular obstruction. PMID- 19555617 TI - Ultrasound validation of maneuvers to increase internal jugular vein cross sectional area and decrease compressibility. AB - OBJECTIVES: The aim of this study is to determine which maneuvers result in greatest cross-sectional area (CSA) of the internal jugular vein (IJV) and reduce collapsibility as measured by ultrasound during simulated venipuncture. METHODS: A total of 52 healthy adult volunteers were prospectively studied in an academic emergency department. Cross-sectional area of the IJV was recorded at baseline, with Valsalva, hepatic pressure, and a combination of hepatic pressure plus Valsalva. Subjects were studied in supine and Trendelenburg. Measurements were repeated using pressure applied to the ultrasound transducer to simulate venipuncture and evaluate degree of IJV collapse. Repeated measures analysis of variance models were used to assess the effects of the maneuvers on the diameter equivalent of the cross-sectional area (CRADE). RESULTS: With simulated venipuncture, both Valsalva and Trendelenburg position were significantly (P < .0001) associated with increased CSA of the IJV. Valsalva in either Trendelenburg or supine position was associated with the largest CRADE (1.20 and 1.13 cm, respectively). Without simulated venipuncture, CSA of the IJV were increased in all settings (P < .0001), but the relative impacts of Valsalva and Trendelenburg position were similar. Hepatic pressure had no impact on CSA of the IJV (P = .94). CONCLUSIONS: All maneuvers with the exception of hepatic pressure led to a statistically significant increase in IJV CSA as compared with baseline with and without simulated venipuncture. Valsalva was the only maneuver, when used alone or in combination, to increase the CSA by greater than 50% and prevent collapse by 50% or more under simulated venipuncture. PMID- 19555618 TI - Head and neck cooling after cardiac arrest results in lower jugular bulb than esophageal temperature. AB - PURPOSE: To determine whether during the initial phase of head and neck cooling, jugular bulb temperature (Tjb; which may reflect brain temperature) is lower than esophageal temperature (Tes). BASIC PROCEDURES: To compare Tes and Tjb, patients received head or head and neck cooling after cardiac arrest. MAIN FINDINGS: The first series with head cooling (n = 5; mean age 54 with a range of 41-62 years; 1 female and 4 males; mean body weight 80 kg with a range of 70-85 kg) showed a mean difference of 0.22 degrees C (95% CI, -1.14 to 0.70; P = .55; limits of agreement, -3.17 to 2.73) between Tes and Tjb over 12 hours. For the second series, with head and neck cooling (n = 6, mean age 65 with a range of 56-76 years; 3 females and 3 males; mean body weight 75 kg with a range of 65-91 kg), Tjb was lower than Tes with a difference of 0.60 degrees C (95% CI, 0.22 to 0.99; P = .01; limits of agreement, -3.10 to 4.30). During the first 3 hours, Tjb decreased faster than Tes (1.1 degrees C/h [95% CI, 0.4 to 1.8; P < .01]). PRINCIPAL CONCLUSION: During the initial phase of therapeutic hypothermia, Tjb seems to be lower than Tes. PMID- 19555619 TI - The Spaso technique: a prospective study of 34 dislocations. AB - BACKGROUND: The Spaso technique has been recently described as a new, simple, and effective manoeuvre for reducing anterior shoulder dislocation. However, there is a lack of series in the English literature. The aim of the study was to evaluate the results obtained with the introduction of this technique in an emergency department. METHODS: An observational prospective study at the Hospital Clinic of Barcelona Orthopaedic Emergency Department was done from January 2007 to May 2007. In this period, the Spaso technique was applied in 36 anterior shoulder dislocations. Two of the patients were excluded due to a presentation later than 24 hours after the dislocation, leaving a total of 34 dislocations in 33 patients. They occurred in 22 male and 12 female shoulders. The patients' ages ranged from 21 to 80 years (average, 51 years). The success of the attempt, complications, and other data were collected. RESULTS: The Spaso technique was successful in 23 (67.6%) of the 34 anterior shoulder dislocations. If just those patients with previous shoulder dislocations were taken into account, the success rate increased to 83%. No complications were observed. CONCLUSION: Based on the results of the present study, we consider the Spaso technique as a safe and effective manoeuvre for the reduction of anterior shoulder dislocations, with an acceptable success rate similar to the classical techniques previously described. PMID- 19555621 TI - Slower infusion of metoclopramide decreases the rate of akathisia. AB - OBJECTIVE: We investigated the difference in incidence of acute akathisia related to the rate of infusion in patients receiving metoclopramide for acute nausea, vomiting, or migraine headache in the emergency department (ED). METHODS: Randomized, prospective, double-blind clinical trial of patients aged 18 years and older who were to receive intravenous metoclopramide for the treatment of nausea, vomiting, or headache were eligible. Patients were excluded if they were taking medications that might mimic or mask akathisia, had a movement disorder, renal insufficiency, or were unable or unwilling to consent. Pregnant women and prisoners were also excluded. Subjects were randomized to receive 1 of 2 accepted metoclopramide administration regimens. The regimens included 10 mg of metoclopramide administered either as a 2-minute bolus (BG) or as a slow infusion for 15 minutes (IG). All patients received a normal saline placebo at the opposite rate to maintain blinding. The main outcome was development of akathisia noted at 60 minutes after drug administration as measured either with The Prince Henry Hospital akathisia rating scale or by sudden unexplained departure from the ED during treatment. RESULTS: One hundred twenty-seven patients were eligible for the study. Fifty-nine patients met exclusion criteria. Of the remaining 68 patients, 36 were randomized to the BG and 32 were randomized to the IG. In the BG, 11.1% of patients developed akathisia compared with 0% in the IG (P = .026). Four patients developed akathisia based on the scale and 2 departed suddenly from the ED. CONCLUSIONS: Slower infusion of metoclopramide reduces the incidence of akathisia. PMID- 19555620 TI - Shortening of cardiopulmonary resuscitation time before the defibrillation worsens the outcome in out-of-hospital VF patients. AB - OBJECTIVE: The purpose of the study is to investigate the influence of cardiopulmonary resuscitation (CPR) time before the first defibrillation. METHODS: The present study retrospectively analyzed the Utstein template records from April 1, 2002, to June 30, 2005. Patients who had out-of-hospital-witnessed cardiac arrest caused by cardiac disease and who presented with ventricular fibrillation (VF) as the initial cardiac rhythm were included in the study. Before April 1, 2003, the emergency medical technician (EMT) needed to obtain telephone permission before attempting defibrillation, and CPR was continued until permission was received (CPR first). On and after April 1, 2003, the EMT was immediately able to attempt a defibrillation without obtaining permission (shock first). RESULTS: In 143 patients who had out-of-hospital-witnessed VF, 43 patients and 100 patients were treated with the CPR-first strategy and the shock first strategy, respectively. The duration of CPR before the first defibrillation was longer in the CPR-first group than that in the shock-first group. The CPR first group showed a higher rate of favorable neurologic outcome 30 days after (28% vs 14%; P = .048) and 1 year after cardiac arrest (26% vs 11%; P = .033) than those of the shock-first group. In the patients with witnessed VF, a stepwise multiple logistic regression analysis showed the CPR-first strategy to improve the neurologic outcome. CONCLUSIONS: In patients with out-of-hospital witnessed VF, sufficient CPR before the first defibrillation is considered to improve the neurologic outcome in comparison to the performance of immediate defibrillation. PMID- 19555622 TI - Intraventricular conduction abnormality--an electrocardiographic algorithm for rapid detection and diagnosis. AB - Intraventricular conduction abnormalities are commonly seen on the electrocardiograms of patients in the emergency department population. The significance of such a finding can vary widely. When placed into clinical context, determination and description of an intraventricular conduction delay (IVCD) can assist in the management of a number of pathologies--directly cardiac and otherwise. The IVCDs discussed in this review include the unifascicular blocks (right bundle branch block [RBBB], left anterior fascicular block [LAFB], and left posterior fascicular block [LPFB]) and the bifascicular blocks (left bundle branch block, RBBB plus LAFB, and RBBB plus LPFB). After a review of these IVCDs, a simplified algorithm that will aid in the electrocardiographic diagnosis of these conduction abnormalities is presented. PMID- 19555623 TI - Bedside ultrasound for verification of shoulder reduction with the lateral and anterior approaches. PMID- 19555624 TI - Management of the 'Can't Intubate, Can't Ventilate' situation in the field. PMID- 19555627 TI - Hypersensitivity myocarditis and hypersensitivity coronary syndrome (Kounis syndrome). PMID- 19555629 TI - Is there sufficient evidence to suggest cyclobenzaprine might be implicated in causing serotonin toxicity? PMID- 19555631 TI - The presentation of 6-mercaptopurine overdose in ED. AB - 6-Mercaptopurine (6-MP), although an effective immunosuppressive when used in the treatment of certain cancers, can have devastating effects when ingested accidentally or used in excessive amounts. We report here the case of an unintentional ingestion of a large amount of 6-MP by a woman with hypothyroidism who was erroneously given this antimetabolic agent by her pharmacist instead the propylthiouracil (PTU) she was actually prescribed. This is one of several documented cases in which 6-MP has been dispensed instead of PTU. Because of the myelosuppressive and hepatotoxic effects of 6-MP, this case reinforces the need for both physicians and patients to understand the importance of examining their medications before ingestion. PMID- 19555632 TI - Middle turbinectomy as a complication of nasopharyngeal airway placement. AB - A nasopharyngeal airway is an adjunct used to relieve airway obstruction due to tongue relaxation in an unconscious or semiconscious patient. It is considered a safe procedure in patients without massive facial trauma or basilar skull fracture. This is a report of a healthy patient undergoing procedural sedation who sustained a middle turbinectomy upon placement of a nasal trumpet to assist ventilation. He subsequently developed severe epistaxis that resolved only after multiple attempts of packing both by the emergency physician as well as the otolaryngology service. It is to be hoped that this will increase awareness among emergency physicians of this unusual complication. PMID- 19555633 TI - Pulmonary embolism presenting as acute abdomen in a child and adult. AB - Pulmonary embolisms (PEs) are easily missed both in children and adults because of the varied presentations and subtle clinical findings. Abdominal pain as the presenting symptom of PE is extremely rare in children and only reported as occasional case reports in adults. We present a series of 2 cases of PE presenting as acute abdomen. Case 1 is a 14-year-old adolescent boy who presented to a pediatric emergency department with abdominal pain, whereas case 2 is a 22 year-old man who presented to the adult emergency department of the same institution with abdominal pain. There was a delay in diagnosis in both cases due to lack of recognition of the unusual presentation. Awareness of the unusual presentations of PE and the risk factors in both adults and children can assist the clinician toward an accurate diagnosis and timely therapeutic intervention. PMID- 19555634 TI - A methicillin-resistant Staphylococcus aureus-positive holospinal epidural abscess. AB - Spinal epidural abscess is a rare debilitating disease that if left untreated may result in serious morbidity and mortality. Most cases involve the level of 3 or 4 vertebrae, but in very rare cases may affect the whole spine. The most common pathogen found in spinal abscesses is Staphylococcus aureus, which involves approximately two thirds of cases. The recent introduction of methicillin resistant strains of S aureus has left physicians with the challenging task of identifying and treating this serious condition.We present the only case reported of a methicillin-resistant S aureus holospinal epidural abscess with subsequent neurological follow-up over a 1-year period. PMID- 19555635 TI - Successful resuscitation of a patient in asystole after a TASER injury using a hypothermia protocol. AB - New studies have shown the benefit of initiating a hypothermia protocol in the survivors of cardiac arrest. Although the data have shown an improved neurologic end point in patients initially in ventricular fibrillation or pulseless ventricular tachycardia, there is still debate about whether patients initially in other rhythms would benefit from hypothermia after return of spontaneous circulation. This is a report of a 17-year-old male found to be in asystole after sustaining a TASER injury, who was treated with a hypothermia protocol after return of spontaneous circulation and left the hospital with intact neurologic function. PMID- 19555636 TI - A 21-year-old man with fever and abdominal pain after recent peritonsillar abscess drainage. AB - Lemierre syndrome an extremely rare and unreported disease typically results from oropharyngeal infection and/or local tissue trauma with invasion of the parapharyngeal soft tissue with bacteria. Once local tissue invasion occurs, internal jugular vein septic thrombophlebitis with or without metastatic complications may occur. The etiology of Lemierre syndrome is generally Fusobacterium necrophorum, reported to be present in approximately 80% of the cases. The outcome of Lemierre syndrome in the preantibiotic era was nearly always fatal. The outcome today with the appropriate antibiotic coverage is quite good. The management of a patient with suspected Lemierre syndrome should be aggressive and comprehensive. Blood cultures as well as cultures from infection sites should be obtained. Computed tomography, magnetic resonance imaging, and ultrasound are all appropriate modalities to detect the presence of internal jugular vein thrombophlebitis. Treatment of Lemierre syndrome consists of prolonged antibiotic therapy (4 to 6 weeks) with appropriate anaerobic coverage. Typical antibiotics include penicillins, metronidazole, or clindamycin. Surgical drainage of metastatic abscesses is often needed ensure resolution of this disease process. Persistent sore throat syndrome caused by F necrophorum has been reported in the literature and the General Practitioners should be aware of this entity because they may see this in their practice. PMID- 19555637 TI - Interhemispheric lipoma masquerading as pneumocranium in a patient with head injury. AB - Head injury is very commonly seen in the emergency department (ED) worldwide, and almost, an unenhanced computed tomography scan of the brain suffices for patients seen in the ED presenting after trauma or with a new neurologic deficit. Intracranial lipomas are usually asymptomatic, and they are often an occasional finding. In head injury cases, lipomas are easily to be misdiagnosed as pneumocranium on brain computed tomography with low-density attenuation image. We presented an extremely rare case of interhemispheric lipoma presenting to an ED as a lesion of acute brain insult. The aim of this report is to emphasize the importance of clinical thinking in the differential diagnosis of markedly hypodense lesion on computed tomography imaging of a patient with head injury. PMID- 19555638 TI - Acute chest syndrome diagnosed by lung sonography. PMID- 19555640 TI - Subacute subdural hematoma in a 45-year-old woman with no significant past medical history after a roller coaster ride. AB - Amusement park ride injuries have been newsworthy events for many years. The multitude and severity of these injuries has been reported many times over the past 20 years and includes spinal cord and vertebral injuries, subarachnoid hemorrhage, internal and vertebral artery dissections, and even a few cases of subdural hematoma (SDH). There has also been as many theories to explain these injuries as there have been injuries themselves including how G forces and rotational acceleration can cause both neuroparenchymal and neurovascular injury. PMID- 19555639 TI - Pituitary adenoma apoplexy with initial presentation mimicking bacterial meningoencephalitis: a case report. AB - Pituitary apoplexy is a rare but life-threatening disorder. Clinical presentation of this condition includes severe headache, impaired consciousness, fever, visual disturbance, and variable ocular paresis. Signs of meningeal irritation are very rare. However, if present and associated with headache, fever, and pleocytosis, meningeal irritation may lead to misinterpretation as infectious meningoencephalitis. To the best of our knowledge, pituitary apoplexy with an initial presentation mimicking infectious meningoencephalitis had rarely been reported in the literature. Here, we report a 57-year-old man who had acute severe headache, high fever, neck stiffness, disturbance in consciousness, and left ocular paresis. Laboratory data showed leukocytosis, an elevated C-reactive protein level, and neutrophilic pleocytosis in the cerebrospinal fluid. Because bacterial meningoencephalitis was suspected, empiric antibiotic therapy was administered but in vain. Further examinations indicated a diagnosis of pituitary adenoma with apoplexy. After the immediate administration of intravenous corticosteroid supplement and surgical decompression, the patient recovered. PMID- 19555641 TI - Reactivating memories for consolidation. AB - The consolidation of memory is thought to occur via a hippocampal-neocortical dialog involving reactivation of memory patterns in the hippocampus during sharp wave ripples. In this issue of Neuron, Nakashiba et al. demonstrate that CA3 output is required for consolidation of contextual fear memory. They also show that lack of CA3 output results in a decrease in ripple-related reactivation, providing additional evidence for a role of ripple-related reactivation in the consolidation process. PMID- 19555642 TI - Tripping the HCN breaker. AB - Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels regulate neuronal excitability, pacemaking, dendritic integration, and homeostatic plasticity and are culprits in aberrant neuronal activity in certain epilepsies. In this issue of Neuron two manuscripts (Santoro et al. and Zolles et al.) report that HCN channel gating and expression are controlled by Trip8b (Pex5R) but with a bidirectional spin. PMID- 19555643 TI - Inhibition acts globally to shape olfactory cortical tuning. AB - Lateral inhibition between near-neighbor neurons has long been thought to be important for narrowing the receptive fields of neurons in many sensory systems. A new study by Poo and Isaacson in this issue of Neuron examining olfactory processing finds that "global" inhibition within the primary olfactory cortex might accomplish a similar end. PMID- 19555644 TI - Applying neuroscience to architecture. AB - Architectural practice and neuroscience research use our brains and minds in much the same way. However, the link between neuroscience knowledge and architectural design--with rare exceptions--has yet to be made. The concept of linking these two fields is a challenge worth considering. PMID- 19555645 TI - Amygdala inhibitory circuits and the control of fear memory. AB - Classical fear conditioning is a powerful behavioral paradigm that is widely used to study the neuronal substrates of learning and memory. Previous studies have clearly identified the amygdala as a key brain structure for acquisition and storage of fear memory traces. Whereas the majority of this work has focused on principal cells and glutamatergic transmission and its plasticity, recent studies have started to shed light on the intricate roles of local inhibitory circuits. Here, we review current understanding and emerging concepts of how local inhibitory circuits in the amygdala control the acquisition, expression, and extinction of conditioned fear at different levels. PMID- 19555646 TI - Graded levels of FGF protein span the midbrain and can instruct graded induction and repression of neural mapping labels. AB - Graded guidance labels are widely used in neural map formation, but it is not well understood which potential strategy leads to their graded expression. In midbrain tectal map development, FGFs can induce an entire midbrain, but their protein distribution is unclear, nor is it known whether they may act instructively to produce graded gene expression. Using a receptor-alkaline phosphatase fusion probe, we find a long-range posterior > anterior FGF protein gradient spanning the midbrain. Heparan sulfate proteoglycan (HSPG) is required for this gradient. To test whether graded FGF concentrations can instruct graded gene expression, a quantitative tectal explant assay was developed. Engrailed-2 and ephrin-As, normally in posterior > anterior tectal gradients, showed graded upregulation. Moreover, EphAs, normally in anterior > posterior countergradients, showed coordinately graded downregulation. These results provide a mechanism to establish graded mapping labels and more generally provide a developmental strategy to coordinately induce a structure and pattern its cell properties in gradients. PMID- 19555647 TI - Hippocampal CA3 output is crucial for ripple-associated reactivation and consolidation of memory. AB - A widely held memory consolidation theory posits that memory of events and space is initially stored in the hippocampus (HPC) in a time-limited manner and is consolidated in the neocortex for permanent storage. Although posttraining HPC lesions result in temporally graded amnesia, the precise HPC circuits and mechanisms involved in remote memory storage remain poorly understood. To investigate the role of the trisynaptic pathway in the consolidation process we employed the CA3-TeTX transgenic mouse, in which CA3 output can be specifically and inducibly controlled. We found that posttraining blockade of CA3 output for up to 4 weeks impairs the consolidation of contextual fear memory. Moreover, in vivo hippocampal recordings revealed a reduced intrinsic frequency of CA1 ripples and a significant decrease in the experience-dependent, ripple-associated coordinated reactivation of CA1 cell pairs. Collectively, these results suggest that the posttraining integrity of the trisynaptic pathway and the ripple associated reactivation of hippocampal memory engram are crucial for memory consolidation. PMID- 19555648 TI - Soluble oligomers of amyloid Beta protein facilitate hippocampal long-term depression by disrupting neuronal glutamate uptake. AB - In Alzheimer's disease (AD), the impairment of declarative memory coincides with the accumulation of extracellular amyloid-beta protein (Abeta) and intraneuronal tau aggregates. Dementia severity correlates with decreased synapse density in hippocampus and cortex. Although numerous studies show that soluble Abeta oligomers inhibit hippocampal long-term potentiation, their role in long-term synaptic depression (LTD) remains unclear. Here, we report that soluble Abeta oligomers from several sources (synthetic, cell culture, human brain extracts) facilitated electrically evoked LTD in the CA1 region. Abeta-enhanced LTD was mediated by mGluR or NMDAR activity. Both forms of LTD were prevented by an extracellular glutamate scavenger system. Abeta-facilitated LTD was mimicked by the glutamate reuptake inhibitor TBOA, including a shared dependence on extracellular calcium levels and activation of PP2B and GSK-3 signaling. In accord, synaptic glutamate uptake was significantly decreased by soluble Abeta. We conclude that soluble Abeta oligomers perturb synaptic plasticity by altering glutamate recycling at the synapse and promoting synapse depression. PMID- 19555649 TI - TRIP8b splice variants form a family of auxiliary subunits that regulate gating and trafficking of HCN channels in the brain. AB - Hyperpolarization-activated cyclic nucleotide-regulated (HCN) channels, which generate the I(h) current, mediate a number of important brain functions. The HCN1 isoform regulates dendritic integration in cortical pyramidal neurons and provides an inhibitory constraint on both working memory in prefrontal cortex and spatial learning and memory in the hippocampus. Altered expression of HCN1 following seizures may contribute to the development of temporal lobe epilepsy. Yet the regulatory networks and pathways governing HCN channel expression and function in the brain are largely unknown. Here, we report the presence of nine alternative N-terminal splice forms of the brain-specific cytoplasmic protein TRIP8b and demonstrate the differential effects of six isoforms to downregulate or upregulate HCN1 surface expression. Furthermore, we find that all TRIP8b isoforms inhibit channel opening by shifting activation to more negative potentials. TRIP8b thus functions as an auxiliary subunit that provides a mechanism for the dynamic regulation of HCN1 channel expression and function. PMID- 19555650 TI - Association with the auxiliary subunit PEX5R/Trip8b controls responsiveness of HCN channels to cAMP and adrenergic stimulation. AB - Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are key modulators of neuronal activity by providing the depolarizing cation current I(h) involved in rhythmogenesis, dendritic integration, and synaptic transmission. These tasks critically depend on the availability of HCN channels, which is dynamically regulated by intracellular cAMP; the range of this regulation, however, largely differs among neurons in the mammalian brain. Using affinity purification and high-resolution mass spectrometry, we identify the PEX5R/Trip8b protein as the beta subunit of HCN channels in the mammalian brain. Coassembly of PEX5R/Trip8b affects HCN channel gating in a subtype-dependent and mode-specific way: activation of HCN2 and HCN4 by cAMP is largely impaired, while gating by phosphoinositides and basal voltage-dependence remain unaffected. De novo expression of PEX5R/Trip8b in cardiomyocytes abolishes beta-adrenergic stimulation of HCN channels. These results demonstrate that PEX5R/Trip8b is an intrinsic auxiliary subunit of brain HCN channels and establish HCN-PEX5R/Trip8b coassembly as a mechanism to control the channels' responsiveness to cyclic nucleotide signaling. PMID- 19555651 TI - Burst-timing-dependent plasticity of NMDA receptor-mediated transmission in midbrain dopamine neurons. AB - Bursts of spikes triggered by sensory stimuli in midbrain dopamine neurons evoke phasic release of dopamine in target brain areas, driving reward-based reinforcement learning and goal-directed behavior. NMDA-type glutamate receptors (NMDARs) play a critical role in the generation of these bursts. Here we report LTP of NMDAR-mediated excitatory transmission onto dopamine neurons in the substantia nigra. Induction of LTP requires burst-evoked Ca2+ signals amplified by preceding metabotropic neurotransmitter inputs in addition to the activation of NMDARs themselves. PKA activity gates LTP induction by regulating the magnitude of Ca2+ signal amplification. This form of plasticity is associative, input specific, reversible, and depends on the relative timing of synaptic input and postsynaptic bursting in a manner analogous to the timing rule for cue-reward learning paradigms in behaving animals. NMDAR plasticity might thus represent a potential neural substrate for conditioned dopamine neuron burst responses to environmental stimuli acquired during reward-based learning. PMID- 19555652 TI - Metaplasticity of hypothalamic synapses following in vivo challenge. AB - Neural networks that regulate an organism's internal environment must sense perturbations, respond appropriately, and then reset. These adaptations should be reflected as changes in the efficacy of the synapses that drive the final output of these homeostatic networks. Here we show that hemorrhage, an in vivo challenge to fluid homeostasis, induces LTD at glutamate synapses onto hypothalamic magnocellular neurosecretory cells (MNCs). LTD requires the activation of postsynaptic alpha2-adrenoceptors and the production of endocannabinoids that act in a retrograde fashion to inhibit glutamate release. In addition, both hemorrhage and noradrenaline downregulate presynaptic group III mGluRs. This loss of mGluR function allows high-frequency activity to potentiate these synapses from their depressed state. These findings demonstrate that noradrenaline controls a form of metaplasticity that may underlie the resetting of homeostatic networks following a successful response to an acute physiological challenge. PMID- 19555654 TI - The brain under self-control: modulation of inhibitory and monitoring cortical networks during hypnotic paralysis. AB - Brain mechanisms of hypnosis are poorly known. Cognitive accounts proposed that executive attentional systems may cause selective inhibition or disconnection of some mental operations. To assess motor and inhibitory brain circuits during hypnotic paralysis, we designed a go-no-go task while volunteers underwent functional magnetic resonance imaging (fMRI) in three conditions: normal state, hypnotic left-hand paralysis, and feigned paralysis. Preparatory activation arose in right motor cortex despite left hypnotic paralysis, indicating preserved motor intentions, but with concomitant increases in precuneus regions that normally mediate imagery and self-awareness. Precuneus also showed enhanced functional connectivity with right motor cortex. Right frontal areas subserving inhibition were activated by no-go trials in normal state and by feigned paralysis, but irrespective of motor blockade or execution during hypnosis. These results suggest that hypnosis may enhance self-monitoring processes to allow internal representations generated by the suggestion to guide behavior but does not act through direct motor inhibition. PMID- 19555655 TI - Export or recombination of charges in reaction centers in intact cells of photosynthetic bacteria. AB - The kinetics and thermodynamics of forward and reverse electron transfer around the reaction center of purple bacterium Rhodobacter sphaeroides were studied in vivo by flash-excited delayed fluorescence, prompt fluorescence (induction) and kinetic difference absorption. By protection of the photomultiplier from intense bacteriochlorophyll prompt fluorescence evoked by laser excitation, the time resolution of the fluorometer was reduced typically 10 micros. Two precursor states of the delayed fluorescence were identified: P(+)Q(A)(-) and cyt c(2)(3+)Q(A)(-) whose enthalpy levels were 340 meV and 1020 meV below A, respectively. The free energy of the P(+)Q(A)(-) state relative to A* was -870 meV in whole cells. Similar values were obtained earlier for isolated reaction center and chromatophore. The free energies of cyt c(2)(3+)Q(A)(-) and P(+)Q(A)( ) states showed no or very weak (-6 meV/pH unit) pH-dependence, respectively, supporting the concept of pH-independent redox midpoint potential of Q(A)/Q(A)(-) in intact cells. In accordance with the multiphasic kinetics of delayed fluorescence, the kinetics of re-opening of the closed reaction center is also complex (it extends up to 1 s) as a consequence of acceptor and donor-side reactions. The control of charge export from the reaction center by light regime, redox agents and inhibitors is investigated. The complex kinetics may arise from the distribution of quinones in different redox states on the acceptor side (Q(B) binding site and pool) and from organization of electron transfer components in supercomplexes. PMID- 19555653 TI - Odor representations in olfactory cortex: "sparse" coding, global inhibition, and oscillations. AB - The properties of cortical circuits underlying central representations of sensory stimuli are poorly understood. Here we use in vivo cell-attached and whole-cell voltage-clamp recordings to reveal how excitatory and inhibitory synaptic input govern odor representations in rat primary olfactory (piriform) cortex. We show that odors evoke spiking activity that is sparse across the cortical population. We find that unbalanced synaptic excitation and inhibition underlie sparse activity: inhibition is widespread and broadly tuned, while excitation is less common and odor-specific. "Global" inhibition can be explained by local interneurons that receive ubiquitous and nonselective odor-evoked excitation. In the temporal domain, while respiration imposes a slow rhythm to olfactory cortical responses, odors evoke fast (15-30 Hz) oscillations in synaptic activity. Oscillatory excitation precedes inhibition, generating brief time windows for precise and temporally sparse spike output. Together, our results reveal that global inhibition and oscillations are major synaptic mechanisms shaping odor representations in olfactory cortex. PMID- 19555656 TI - Novel A14841G mutation is associated with high penetrance of LHON/C4171A family. AB - We report the clinical and genetic characterization of a Chinese LHON family carrying an ND1/C4171A mutation. This family has high penetrance of visual impairment and extremely low frequency of vision recovery, which is in marked contrast to previously reported results for Korean LHON families with the same mutation. Sequence analysis of the complete mtDNA in the partially defined East Asian haplogroup N9a1 revealed the presence of 29 other variants. A novel heteroplasmic A14841G mutation, one of the variants with a serine substituted for a highly conserved asparagine at amino acid 32 of Cytochrome b (Cytb), may play a synergistic role with the C4171A mutation, leading to significantly different clinical manifestations of LHON among these families. The study further confirmed that C4171A was a rare primary LHON mutation, and the mtDNA background could also contribute to the clinical manifestation of the LHON/C4171A mutation. PMID- 19555657 TI - Dynamic hydrostatic pressure promotes differentiation of human dental pulp stem cells. AB - The masticatory apparatus absorbs high occlusal forces, but uncontrolled parafunctional or orthodontic forces damage periodontal ligament (PDL), cause pulpal calcification, pulp necrosis and tooth loss. Morphology and functional differentiation of connective tissue cells can be controlled by mechanical stimuli but effects of uncontrolled forces on intra-pulpal homeostasis and ability of dental pulp stem cells (DPSCs) to withstand direct external forces are unclear. Using dynamic hydrostatic pressure (HSP), we tested the hypothesis that direct HSP disrupts DPSC survival and odontogenic differentiation. DPSCs from four teenage patients were subjected to HSP followed by assessment of cell adhesion, survival and recovery capacity based on odontogenic differentiation, mineralization and responsiveness to bone morphogenetic protein-2 (BMP-2). HSP down-regulated DPSC adhesion and survival but promoted differentiation by increasing mineralization, in vivo hard tissue regeneration and BMP-2 responsiveness despite reduced cell numbers. HSP-treated DPSCs displayed enhanced odontogenic differentiation, an indication of favorable recovery from HSP-induced cellular stress. PMID- 19555658 TI - Structures of native human thymidine phosphorylase and in complex with 5 iodouracil. AB - Thymidine phosphorylase (TP) first identified as platelet derived endothelial cell growth factor (PD-ECGF) plays a key role in nucleoside metabolism. Human TP (hTP) is implicated in angiogenesis and is overexpressed in several solid tumors. Here, we report the crystal structures of recombinant hTP and its complex with a substrate 5-iodouracil (5IUR) at 3.0 and 2.5A, respectively. In addition, we provide information on the role of specific residues in the enzymatic activity of hTP through mutagenesis and kinetic studies. PMID- 19555659 TI - Titanium dioxide nanoparticles induce apoptosis through the JNK/p38-caspase-8-Bid pathway in phytohemagglutinin-stimulated human lymphocytes. AB - We investigated the signaling pathways underlying nano-TiO(2)-induced apoptosis in cultured human lymphocytes. Nano-TiO(2) increased the proportion of sub-G1 cells, activated caspase-9 and caspase-3, and induced caspase-3-mediated PARP cleavage. Nano-TiO(2) also induced loss of mitochondrial membrane potential, which suggests that nano-TiO(2) induces apoptosis via a mitochondrial pathway. A time-sequence analysis of the induction of apoptosis by nano-TiO(2) revealed that nano-TiO(2) triggered apoptosis through caspase-8/Bid activation. We also observed that inhibition of caspase-8 by z-IETD-fmk suppressed the caspase-8/Bid activation, caspase-3-mediated PARP cleavage, and apoptosis. Nano-TiO(2) activated two MAPKs, p38 and JNK. In addition, the selective p38 inhibitor SB203580 and selective JNK inhibitor SP600125 suppressed nano-TiO(2)-induced apoptosis and caspase-8 activation to moderate and significant extents, respectively. Knockdown of protein levels of JNK1 and p38 using an RNA interference technique also suppressed caspase-8 activation. Our results suggest that nano-TiO(2)-induced apoptosis is mediated by the p38/JNK pathway and the caspase-8-dependent Bid pathway in human lymphocytes. PMID- 19555660 TI - Specific roles of Rac1 and Rac2 in motile functions of HT1080 fibrosarcoma cells. AB - Rho family proteins are constitutively activated in the highly invasive human fibrosarcoma HT1080 cells. We now investigated the specific roles of Rac1 and Rac2 in regulating morphology, F-actin organization, adhesion, migration, and chemotaxis of HT1080 cells. Downregulation of Rac1 using specific siRNA probes resulted in cell rounding, markedly decreased spreading, adhesion, and chemotaxis of HT1080 cells. 2D migration on laminin-coated surfaces in contrast was not markedly affected. Selective Rac2 depletion did not affect cell morphology, cell adhesion, and 2D migration, but significantly reduced chemotaxis. Downregulation of both Rac1 and Rac2 resulted in an even more marked reduction, but not complete abolishment, of chemotaxis indicating distinct as well as overlapping roles of both proteins in chemotaxis. Rac1 thus is selectively required for HT1080 cell spreading and adhesion whereas Rac1 and Rac2 are both required for efficient chemotaxis. PMID- 19555661 TI - Beta2-strand of salivary S cystatins: a "chemeleon sequence". AB - Secondary structure prediction of salivary cystatins S, SA, and SN carried out by several methods label the 39-58 sequence (beta2-strand) as predominantly alpha helical. The helical propensity of a peptide corresponding to beta2-strand of salivary SA cystatin analyzed by CD display high helical propensity in aqueous solution, whereas peptides matching the beta2-strand amino acid sequence of cystatins S and SN, display random coil conformation in aqueous solution but acquire alpha-helical conformation in the presence of trifluoroethanol (TFE). Moreover molecular dynamics simulation performed on the homology modeling of cystatin SA constructed on the basis of recently determined three-dimensional structure of salivary cystatin D, suggests that cystatin SA does not significantly deviate from the starting structure over the course of the simulation. The results obtained indicate that the beta2-strand of salivary S cystatins has high helical propensity when isolated from native protein and acquire the final beta structure by interaction with the rest of the polypeptide chain. PMID- 19555662 TI - Transcriptional activation of NAD+-dependent protein deacetylase SIRT1 by nuclear receptor TLX. AB - An orphan nuclear receptor TLX is a transcriptional repressor that promotes the proliferation and self-renewal of neural precursor cells (NPCs). SIRT1, an NAD(+) dependent protein deacetylase, is highly expressed in the NPCs and participates in neurogenesis. Here, we found that TLX colocalized with SIRT1 and knockdown of TLX by small interfering RNAs decreased SIRT1 levels in NPCs. TLX increased the SIRT1 expression by binding to the newly identified TLX-activating element in the SIRT1 gene promoter in HEK293 cells. Thus, TLX is an inducer of SIRT1 and may contribute to neurogenesis both as a transactivator and as a repressor. PMID- 19555663 TI - Heat shock protein 70 is secreted from endothelial cells by a non-classical pathway involving exosomes. AB - Emerging evidence suggests that a high level of circulating heat shock protein 70 (HSP70) correlates with a lower risk of vascular disease; however, the biological significance of this inverse relationship has not been explored. Herein, we report that oxidative low density lipoprotein (Ox-LDL) and homocysteine (Hcy) induce HSP70 release from endothelial cells. In rat endothelial cells, Ox-LDL and Hcy induced robust release of HSP70, independent of the classical route of endoplasmic reticulum/Golgi protein trafficking or the formation of lipid rafts. In contrast, Ox-LDL and Hcy significantly enhanced the exosomal secretory rate and increased the HSP70 content of exosomes. Exogenous HSP70 had no impact on LPS , Ox-LDL- and Hcy-induced activation of endothelial cells, whereas HSP70 did activate monocytes alone, resulting in monocyte adhesion to endothelial cells. These results indicate that exosome-dependent secretion of HSP70 from endothelial cells provides a novel paracrine mechanism to regulate vascular endothelial functional integrity. PMID- 19555664 TI - Thioredoxin reductase 1 upregulates MCP-1 release in human endothelial cells. AB - To know if thioredoxin reductase 1 (TrxR1) plays a role in antioxidant defense mechanisms against atherosclerosis, effect of TrxR1 on expression/release of monocyte chemoattractant protein (MCP-1) was investigated in activated human endothelial-like EAhy926 cells. The MCP-1 release and expression, cellular generation of reactive oxygen species (ROS), nuclear translocation and DNA binding activity of NF-kappaB subunit p65 were assayed in cells either overexpressing recombinant TrxR1 or having their endogenous TrxR1 knocked down. It was found that overexpression of TrxR1 enhanced, while knockdown of TrxR1 reduced MCP-1 release and expression. Upregulation of MCP-1 by TrxR1 was associated with increasing generation of intracellular ROS generation, enhanced nuclear translocation and DNA-binding activity of NF-kappaB. Assay using NF kappaB reporter revealed that TrxR1 upregulated transcriptional activity of NF kappaB. This study suggests that TrxR1 enhances ROS generation, NF-kappaB activity and subsequent MCP-1 expression in endothelial cells, and may promote rather than prevent vascular endothelium from forming atherosclerotic plaque. PMID- 19555665 TI - NKG2D and CD94 bind to heparin and sulfate-containing polysaccharides. AB - Killer lectin-like receptors NKG2D and CD94 on natural killer cells trigger cytotoxicity through binding of glycans on target cells including sialyl Lewis X antigen. We previously reported that NKG2D and CD94 recognize alpha2,3-linked NeuAc on multi-antennary N-glycans. Here we further investigated polysaccharide binding by these receptors, using glutathione-S-transferase-fused extracellular domains of NKG2D AA 73-216 (rNKG2Dlec) and CD94 AA 68-179 (rCD94lec). We found that rNKG2Dlec and rCD94lec bind in a dose-dependent manner to plates coated with heparin-conjugated bovine serum albumin (heparin-BSA). Binding to heparin-BSA was suppressed by soluble sulfate-containing polysaccharides, but minimally impacted by 2-O-, 6-O-, and 2-N-desulfated heparin. Mutagenesis revealed that (152)Y and (199)Y of NKG2D and (144)F, (160)N, and (166)C of CD94 were critical for binding to heparin-BSA. The present manuscript provides the first evidence that NKG2D and CD94 bind to heparin and sulfate-containing polysaccharides. PMID- 19555666 TI - Roles of proteasomal 19S regulatory particles in promoter loading of thyroid hormone receptor. AB - 19S regulatory particles (19SRP) of 26S proteasome participate in multiple steps of gene transcription in yeast. We previously showed that Tat-binding protein-1 (TBP-1), an ATPase of 19SRP, interacts with thyroid hormone receptor (TR) and enhances TR-mediated transcription synergistically with steroid receptor coactivator-1 (SRC-1). To further elucidate the roles of ATPases and a non-ATPase component of 19SRP in gene regulation by TR, we investigated whether knockdown (KO) of TBP-1, TRIP1 or Rpn10 using small interfering RNA affects TR-mediated transactivation in HeLa cells. KO of individual subunits attenuated TR-mediated transactivation through the thyroid hormone response element (TRE) in the absence or presence of cotransfected SRC-1 without altering TR and SRC-1 protein levels. KO of TBP-1 disrupted ligand-induced loading of TR, SRC-1, and RNA polymerase II in chromatin immunoprecipitation assays. Collectively, both ATPase and non-ATPase components of 19SRP play critical roles in TR-mediated transactivation by coordinating the proper loading of liganded TR to TRE. PMID- 19555667 TI - Specific regulation of protein phosphatase 2A PR72/B'' subunits by calpain. AB - Protein phosphatase 2A (PP2A) represents a family of multimeric serine/threonine phosphatases with pleiotropic roles in signal transduction. We previously described the functional analysis of two Ca(2+)-binding EF-hands in the PR72/B'' class of regulatory PP2A subunits. Now we report partial degradation of PR72/B"alpha2 and PR130/B"alpha1 into a 45-48kDa proteolysis-resistant fragment ('PR45') by the Ca(2+)-dependent protease m-calpain. This limited proteolysis is dependent on EF-hand integrity, independent of two PEST-domains, and highly specific as PP2A(C), PR65/A and representatives of PR55/B and PR61/B' subunit families are calpain-resistant. PR45 was also generated in staurosporine-induced apoptotic MCF7 cells in a calpain-dependent way. Calpain treatment weakens the PR72-core enzyme interaction, activates basal PP2A(T72) phosphatase activity and dramatically increases its sensitivity for and activation by polycations. This unique property can be exploited in a specific biochemical assay for these holoenzymes. We propose local calpain action in vivo may constitute a novel regulatory mechanism of these holoenzymes. PMID- 19555668 TI - Stimulation of dopamine release by nicotinic acetylcholine receptor ligands in rat brain slices correlates with the profile of high, but not low, sensitivity alpha4beta2 subunit combination. AB - alpha4beta2 neuronal nicotinic receptors (nAChRs) can exist in high and low sensitivity states possibly due to distinct stoichiometries during subunit assembly: (alpha4)(2)(beta2)(3) pentamer (high sensitivity, HS) and (alpha4)(3)(beta2)(2) pentamer (low sensitivity, LS). To determine if there is a linkage between HS or LS states and receptor-mediated responses in brain, we profiled several clinically studied alpha4beta2* nAChR agonists for the displacement of radioligand binding to alpha4beta2 [(3)H]-cytisine sites in rat brain membranes, effects on stimulation of [(3)H]-dopamine release from slices of rat prefrontal cortex and striatum, and activation of HS and LS human alpha4beta2 nAChRs expressed in Xenopus laevis oocytes. Binding affinities (pK(i)) and potency (pEC(50)) values for [(3)H]-dopamine release closely correlated with a rank order: varenicline>(-)-nicotine>AZD3480>dianicline congruent with ABT-089. Further, a good correlation was observed between [(3)H]-dopamine release and HS alpha4beta2 pEC(50) values, but not between [(3)H]-dopamine release and LS alpha4beta2. The relative efficacies of the agonists ranged from full to partial agonists. Varenicline behaved as a partial agonist in stimulating [(3)H]-dopamine release and activating both HS and LS alpha4beta2 nAChRs expressed in oocytes. Conversely, ABT-089, AZD3480 and dianicline exhibited little efficacy at LS alpha4beta2 (<5%), were more effective at HS alpha4beta2 nAChRs, and in stimulating cortical and striatal [(3)H]-dopamine release >or=30%. In the presence of alpha-conotoxin MII to block alpha6beta2* nAChRs, the alpha4beta2* alpha-conotoxin-insensitive [(3)H]-dopamine release stimulated by these ligands correlates well with their interactions at HS, but not LS. In summary, this study provides support for HS alpha4beta2* nAChR involvement in neurotransmitter release. PMID- 19555669 TI - Transcriptional down-regulation of Bcl-2 by vinorelbine: identification of a novel binding site of p53 on Bcl-2 promoter. AB - The Bcl-2 family contains a panel of proteins which are conserved regulators of apoptosis in mammalian cells, like the anti-apoptotic protein Bcl-2. According to its significant role in altering susceptibility to apoptosis, the deciphering of the mechanism of Bcl-2 expression modulation may be crucial for identifying therapeutics strategies for cancer. Treatment with microtubule-targeting agents, including taxanes and Vinca alkaloids, generally leads to a decrease in Bcl-2 intracellular amounts. Whereas the interest for these chemotherapeutics is accompanied by advances in the fundamental understanding of their anticancer properties, the molecular mechanism underlying changes in Bcl2 expression remains poorly understood. We report here that p53 contributes to vinorelbine-induced Bcl 2 down-regulation. Indeed, the decrease in Bcl-2 protein levels observed during vinorelbine-induced apoptosis was correlated to the decrease in mRNA levels, as a result of the inhibition of Bcl-2 transcription and promoter activity. In this context, we evaluated p53 contribution in the Bcl-2 transcriptional down regulation. We identified, by chromatin immunoprecipitation, a novel p53 binding site in the Bcl-2 promoter, within a region upstream P(1) promoter. We showed that vinorelbine treatment increased this interaction in A549 cells. This work strengthens the links between p53 and Bcl-2 at a transcriptional level, upon microtubule-targeting agent treatment. Our study also provides answers that will be useful to assess microtubule-targeting agents' mechanism of action and that may help to better understand and increase their effectiveness. PMID- 19555670 TI - Concomitant downregulation of proliferation/survival pathways dependent on FGF R3, JAK2 and BCMA in human multiple myeloma cells by multi-kinase targeting. AB - The identification of proliferation/survival pathways constitutively activated by genetic alterations in multiple myeloma (MM), or sustained by the bone marrow (BM) microenvironment, provides novel opportunities for the development of targeted therapies. The deregulated function of protein tyrosine kinases plays a critical role in driving MM malignant phenotype. We investigated the effects of the multi-target tyrosine kinase inhibitor RPI-1 in a panel of human MM cell lines, including t(4;14) positive cell lines expressing the TK receptor FGF-R3. Cells harboring FGF-R3 activating mutations (KMS11 and OPM2) displayed the highest sensitivity to RPI-1 antiproliferative effect. The stimulating effect of the aFGF ligand was abrogated in cells harboring a non-constitutively active receptor. Drug treatment inhibited activation and expression of the FGF-R3(Y373C) mutant as well as aFGF-dependent signaling involving AKT and ERKs. Inhibition of JAK2, an additional RPI-1 target, resulted in STAT3 inactivation. Blockade of these proliferation/survival pathways was associated with caspase-dependent apoptosis. Moreover, drug treatment abrogated proliferative and pro-invasive stimuli provided by conditioned medium from mesenchymal stromal cells. Gene expression profile of KMS11 cells showed 22 upregulated and 52 downregulated genes upon RPI-1 treatment, with an early modulation of genes implicated in MM pathobiology such as SAT-1, MYC, MIP-1alpha/beta, FGF-R3, and the growth factor receptor B-cell maturation antigen (BCMA). Thus, concomitant blockade of FGF-R3 and JAK2 results in inhibition of several MM-promoting pathways, including BCMA regulated signaling, and downregulation of disease-associated proteins. These data may have therapeutic implications in the design of treatment strategies resulting in the concomitant inhibition of FGF-R3 and JAK2 signaling pathways in t(4;14) MM. PMID- 19555671 TI - The plant sterol guggulsterone attenuates inflammation and immune dysfunction in murine models of inflammatory bowel disease. AB - Inflammatory bowel diseases (IBD) are chronic inflammatory and relapsing diseases of the gut that may manifest as either Crohn's disease (CD) or ulcerative colitis (UC). CD and UC are immunologically different diseases characterized by exacerbated Th1 and Th2 response. T-cell resistance against apoptosis contributes to inappropriate T-cell accumulation and the perpetuation of chronic mucosal inflammation. In the present study we have investigated the effect exerted by guggulsterone (GS) a plant derived steroid isolated from the gum resin of the Commiphora mukul tree, in two models of intestinal inflammation induced in mice by trinitro-benzene sulfonic acid (TNBS) and oxazolone. We provided evidence that E-GS protects mice against development of sign and symptoms of colon inflammation. E-GS effectively attenuated the severity of wasting disease and the fecal score and colon inflammation as assessed by measuring the macroscopic- and microscopic-damage scores. Administration Z-GS failed to ameliorate colon inflammation in TNBS-induced colitis and had a partial effect in oxazolone induced colitis. In vitro, mechanistic studies carried out using CD4+ cells isolated from the intestinal lamina propria demonstrate that GS effectively regulates the function of effector T cells by modulating cell signaling activation pathway caused by CD3/CD28. The net biological effects resulting from exposure to GS includes attenuation of generation of interleukin-2 and -4 and interferon-gamma as well as T cell proliferation. In conclusion, GS is an anti inflammatory compound with the capacity to prevent and ameliorate T-cell-induced colitis. These data ground the use of GS, a natural cholesterol-lowering agent, in the treatment of chronic inflammatory diseases. PMID- 19555672 TI - Prostaglandins compromise basal forebrain cholinergic neuron differentiation and survival: action at EP1/3 receptors results in AIF-induced death. AB - Activated microglia produce a factor or cocktail of factors that promotes cholinergic neuronal differentiation of undifferentiated precursors in the embryonic basal forebrain (BF) in vitro. To determine whether microglial prostaglandins mediate this action, microglia were stimulated in the presence of the cyclooxygenase inhibitor ibuprofen, and microglial conditioned medium (CM) was used to culture rat BF precursors at embryonic day 15. Choline acetyltransferase (ChAT) activity served as a measure of cholinergic differentiation. While inhibition of prostaglandin biosynthesis did not affect the ability of microglial CM to promote ChAT activity, treatment of microglia with prostaglandin E2 (PGE2) inhibited it. Agonists of E prostanoid receptors EP2 (butaprost) and EP1/3 (sulprostone) mimicked PGE2, while misoprostol (E1-4) actually enhanced the action of CM. PGE2 added directly to BF cultures together with microglial CM also inhibited ChAT activity. While BF cultures expressed all four prostanoid receptors, direct addition of sulprostone but not butaprost mimicked PGE2, suggesting that PGE2 engaged EP1/3 receptors in the BF. Neither PKA inhibition by H89 nor cAMP induction by forskolin or dibutyrl-cAMP altered the action of sulprostone. Sulprostone severely compromised ChAT activity, dendrite number, axonal length and axonal branching, but caspase inhibition did not restore these. However, sulprostone resulted in increased staining intensity and nuclear translocation of apoptosis-inducing factor (AIF) suggesting caspase independent cell death. We have found that PGE2 action at microglial EP2 receptors inhibits the microglial production of the cholinergic differentiating cocktail, while action at neuronal EP3 receptors has a deleterious effect on cholinergic neurons causing neurite retraction and cell death. PMID- 19555674 TI - Neural correlates of processing stressful information: an event-related fMRI study. AB - Recent neuroimaging studies investigating neural correlates of psychological stress employ cognitive paradigms that induce a significant hormonal stress response in the scanner. The Montreal Imaging Stress Task (MIST) is one such task that combines challenging mental arithmetic with negative social evaluative feedback. Due to the block design nature of the MIST, it has not been possible thus far to investigate which brain areas respond specifically to the key components of the MIST (mental arithmetic, failure, negative social evaluation). In the current study, we developed an event-related MIST (eventMIST) in order to investigate which neural activation patterns are associated with performing mental arithmetic vs. processing of social evaluative threat. Data was available from twenty healthy university students. The eventMIST induced a significant stress response in a subsample of subjects, called the responders (n=7). Direct comparison between brain activity changes in responders vs. non-responders, in response to challenging math, revealed increased activity bilaterally in dorsomedial prefrontal cortex (PFC), left temporal pole, and right dorsolateral PFC. In response to negative social evaluation, responders showed reduction of brain activity in limbic system regions (medial orbitofrontal cortex and hippocampus), which was largely lacking in non-responders. Direct comparison between the groups for this contrast did not reveal any significant difference, probably due to small number of events available. This is the first study to use an event-related paradigm to investigate brain activity patterns in relation to challenging math and social evaluative threat separately. PMID- 19555673 TI - Exercise-induced improvement in cognitive performance after traumatic brain injury in rats is dependent on BDNF activation. AB - We have previously shown that voluntary exercise upregulates brain derived neurotrophic factor (BDNF) within the hippocampus and is associated with an enhancement of cognitive recovery after a lateral fluid percussion injury (FPI). In order to determine if BDNF is critical to this effect we used an immunoadhesin chimera (TrkB-IgG) that inactivates free BDNF. This BDNF inhibitor was administered to adult male rats two weeks after they had received a mild fluid percussion injury (FPI) or sham surgery. These animals were then housed with or without access to a running wheel (RW) from post-injury-day (PID) 14 to 20. On PID 21, rats were tested for spatial learning in a Morris Water Maze. Results showed that exercise counteracted the cognitive deficits associated with the injury. However this exercise-induced cognitive improvement was attenuated in the FPI-RW rats that were treated with TrkB-IgG. Molecules important for synaptic plasticity and learning were measured in a separate group of rats that were sacrificed immediately after exercise (PID 21). Western blot analyses showed that exercise increased the mature form of BDNF, synapsin I and cyclic-AMP response element-binding protein (CREB) in the vehicle treated Sham-RW group. However, only the mature form of BDNF and CREB were increased in the vehicle treated FPI RW group. Blocking BDNF (pre administration of TrkB-IgG) greatly reduced the molecular effects of exercise in that exercise-induced increases of BDNF, synapsin I and CREB were not observed. These studies provide evidence that BDNF has a major role in exercise's cognitive effects in traumatically injured brain. PMID- 19555676 TI - Effects of N-acetyl-cysteine on the survival and regeneration of sural sensory neurons in adult rats. AB - Microsurgical reconstruction of injured peripheral nerves often results in limited functional recovery. One contributing factor is the retrograde neuronal degeneration of sensory neurons in the dorsal root ganglia (DRG) and of motor neurons in the spinal cord. The present study investigates the neuroprotective and growth-promoting effects of N-acetyl-cysteine (NAC) on sensory DRG neurons and spinal motoneurons after sciatic axotomy and nerve grafting in adult rats. Sciatic axotomy and nerve grafting were performed at 1 week after sural DRG neurons and motoneurons were retrogradely labeled with the fluorescent tracer Fast Blue. To assess the efficacy of axonal regeneration, a second fluorescent dye Fluoro-Ruby was applied distal to the graft at 12 weeks after nerve repair. At 8-13 weeks after axotomy, only 52-56% of the sural sensory neurons remained in the lumbar DRG, while the majority of motoneurons survived the sciatic nerve injury. Nerve grafting alone or continuous intrathecal NAC treatment (2.4 mg/day) improved survival of sural DRG neurons. Combined treatment with nerve graft and NAC had significant additive effect on neuronal survival and also increased the number of sensory neurons regenerating across the graft. However, NAC treatment neither affected the number of regenerating motoneurons nor the number of myelinated axons in the nerve graft or in the distal nerve stump. The present results demonstrate that NAC provides a highly significant effect of neuroprotection in an animal nerve injury model and that combination with nerve grafting further attenuates retrograde cell death and promotes regeneration of sensory neurons. PMID- 19555675 TI - Spinal translocator protein (TSPO) modulates pain behavior in rats with CFA induced monoarthritis. AB - Translocator protein 18 kDa (TSPO), previously known as the peripheral benzodiazepine receptor (PBR), is predominantly located in the mitochondrial outer membrane and plays an important role in steroidogenesis, immunomodulation, cell survival and proliferation. Previous studies have shown an increased expression of TSPO centrally in neuropathology, as well as in injured nerves. TSPO has also been implicated in modulation of nociception. In the present study, we examined the hypothesis that TSPO is involved in the initiation and maintenance of inflammatory pain using a rat model of Complete Freund's Adjuvant (CFA)-induced monoarthritis of the tibio-tarsal joint. Immunohistochemistry was performed using Iba-1 (microglia), NeuN (neurons), anti-Glial Fibrillary Acidic Protein, GFAP (astrocytes) and anti-PBR (TSPO) on Days 1, 7 and 14 after CFA induced arthritis. Rats with CFA-induced monoarthritis showed mechanical allodynia and thermal hyperalgesia on the ipsilateral hindpaw, which correlated with the increased TSPO expression in ipsilateral laminae I-II on all experimental days. Iba-1 expression in the ipsilateral dorsal horn was also increased on Days 7 and 14. Moreover, TSPO was colocalized with Iba-1, GFAP and NeuN within the spinal cord dorsal horn. The TSPO agonist Ro5-4864, given intrathecally, dose-dependently retarded or prevented the development of mechanical allodynia and thermal hyperalgesia in rats with CFA-induced monoarthritis. These findings provide evidence that spinal TSPO is involved in the development and maintenance of inflammatory pain behaviors in rats. Thus, spinal TSPO may present a central target as a complementary therapy to reduce inflammatory pain. PMID- 19555677 TI - Repeated exposure to MDMA provides neuroprotection against subsequent MDMA induced serotonin depletion in brain. AB - Repeated exposure to sub-lethal insults has been reported to result in neuroprotection against a subsequent deleterious insult. The purpose of this study was to evaluate whether repeated exposure (preconditioning) to a non-5-HT depleting dose of MDMA in adult rats provides neuroprotection against subsequent MDMA-induced 5-HT depletion. Treatment of rats with MDMA (10 mg/kg, ip every 2 h for 4 injections) resulted in a 50-65% depletion of 5-HT in the striatum, hippocampus and cortex, and these depletions were significantly attenuated in rats that received a preconditioning regimen of MDMA (10 mg/kg, ip daily for 4 days). The 5-HT depleting regimen of MDMA also resulted in a 40-80% reduction in 5-HT transporter immunoreactivity (SERT(ir)), and the reduction in SERT(ir) also was completely attenuated in MDMA-preconditioned animals. Preconditioning with MDMA (10 mg/kg, ip) daily for 4 days provided neuroprotection against methamphetamine-induced 5-HT depletion, but not dopamine depletion, in the striatum. Additional studies were conducted to exclude the possibility that alterations in MDMA pharmacokinetics or MDMA-induced hyperthermia in rats previously exposed to MDMA contribute towards neuroprotection. During the administration of the 5-HT depleting regimen of MDMA, there was no difference in the extracellular concentration of the drug in the striatum of rats that had received 4 prior, daily injections of vehicle or MDMA. Moreover, there was no difference in the hyperthermic response to the 5-HT depleting regimen of MDMA in rats that had earlier received 4 daily injections of vehicle or MDMA. Furthermore, hyperthermia induced by MDMA during preconditioning appears not to contribute towards neuroprotection, inasmuch as preconditioning with MDMA at a low ambient temperature at which hyperthermia was absent did not alter the neuroprotection provided by the preconditioning regimen. Thus, prior exposure to MDMA affords protection against the long-term depletion of brain 5-HT produced by subsequent MDMA administration. The mechanisms underlying preconditioning-induced neuroprotection for MDMA remain to be determined. PMID- 19555679 TI - Gastroenterology year in review. PMID- 19555678 TI - Comparison of quercetin and dihydroquercetin: antioxidant-independent actions on erythrocyte and platelet membrane. AB - We investigated the effects of two flavonoids quercetin and dihydroquercetin (DHQ), which have different solubilities and antioxidant capacities, on hemolysis and platelet aggregation in human blood. Exposure of human red blood cells (RBCs) to free radicals generated by 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) for 2h resulted in 63.5+/-3.9% hemolysis (vehicle: 0.3+/-0.4%). Pre incubation of RBCs with lipid-soluble quercetin and water-soluble DHQ for 30min significantly reduced the AAPH-induced hemolysis to 3.6+/-1.5% and 32.5+/-5.6% respectively. In contrast, quercetin and DHQ were similarly effective in reducing phospholipase C-induced hemolysis (37.2+/-9.1% and 45.4+/-10.0% versus vehicle 75.7+/-5.2%, P<0.001). Pre-incubation with quercetin, but not DHQ, inhibited the aggregation of platelets by adenosine diphosphate. DHQ was more potent than quercetin in inhibiting superoxide produced by xanthine oxidase. These results suggest that the antihemolytic effects of flavonoids may not be directly mediated by removal of free radicals and may likely be due to their interaction with cell membrane. PMID- 19555681 TI - The potential beneficial effects of beta adrenergic blockade in the treatment of ventricular fibrillation. AB - Cardiac arrest remains a major medical emergency in Western societies, with ventricular fibrillation being the initial rhythm in a significant proportion of cases. Adrenaline is generally accepted to improve the resuscitation outcome, since it improves coronary and cerebral blood flow during cardiopulmonary resuscitation, but several detrimental effects have been associated with its use, most of which are thought to be mediated by its beta adrenergic properties. Several animal studies suggest that beta adrenergic blockade during resuscitation, is associated with increased rates of resuscitation and improved post-resuscitation myocardial function. This article reviews the presence and function of beta-adrenoceptor subtypes in the intact and diseased human myocardium, as well as the differences observed in beta(1)- and beta(2) adrenoceptor subtypes in different species. PMID- 19555680 TI - Dazap2 is required for FGF-mediated posterior neural patterning, independent of Wnt and Cdx function. AB - The organization of the embryonic neural plate requires coordination of multiple signal transduction pathways, including fibroblast growth factors (FGFs), bone morphogenetic proteins (BMPs), and WNTs. Many studies have suggested that a critical component of this process is the patterning of posterior neural tissues by an FGF-caudal signaling cascade. Here, we have identified a novel player, Dazap2, and show that it is required in vivo for posterior neural fate. Loss of Dazap2 in embryos resulted in diminished expression of hoxb9 with a concurrent increase in the anterior marker otx2. Furthermore, we found that Dazap2 is required for FGF dependent posterior patterning; surprisingly, this is independent of Cdx activity. Furthermore, in contrast to FGF activity, Dazap2 induction of hoxb9 is not blocked by loss of canonical Wnt signaling. Functionally, we found that increasing Dazap2 levels alters neural patterning and induces posterior neural markers. This activity overcomes the anteriorizing effects of noggin, and is downstream of FGF receptor activation. Our results strongly suggest that Dazap2 is a novel and essential branch of FGF-induced neural patterning. PMID- 19555682 TI - Blood-brain barrier breakdown by PAF and protection by XQ-1H due to antagonism of PAF effects. AB - Hypoxia and reoxygenation set in motion a series of events, including blood-brain barrier breakdown. We examined the content and effect of platelet-activating factor (PAF), which was increased in the rat brain microvessel endothelial cells (RBMECs) during hypoxia and reoxygenation. MTT method was used to assay cell damage; ELISA analysis was used to estimate PAF release after hypoxia and reoxygenation injury; and RT-PCR and Western blotting method were used to assess gene and protein expressions of inducible nitric-oxide synthase (iNOS) in RBMECs under PAF damage. PAF affected intracellular free Ca(2+) levels, increasing [Ca(2+)](i), which caused up-regulation of iNOS. We also examined the blood-brain barrier protective effect of XQ-1H, a novel ginkgolide B derivative. Pretreatment with XQ-1H (10 microM and 3 microM) for 24 h significantly antagonized PAF receptor and inhibited the increase in intracellular calcium concentration and the up-regulation of iNOS in response to PAF under hypoxia and reoxygenation in vitro. PMID- 19555683 TI - Renal ischemia/reperfusion injury in rats is attenuated by a synthetic glycine derivative. AB - Renal ischemia/reperfusion is a common cause of acute renal failure. Glycine is an effective anti-inflammatory, cytoprotective agent and is reported to have a beneficial effect against ischemia/reperfusion injury in various organs. Previous research notes that free radicals and inflammatory leukocytes both play important roles in the pathogenesis of renal ischemia/reperfusion injury. To develop new therapeutic agents against renal ischemia/reperfusion injury, we sought to link an antioxidant moiety (nitronyl nitroxide) to glycine in the hope that the resulting glycine-nitronyl nitroxide conjugate (GNN) would provide a synergetic protection against renal ischemia/reperfusion injury. In this manuscript, we report the synthesis and biological evaluation of the GNN conjugate. The biological activity of the GNN conjugate was evaluated in an in vivo rat model of renal ischemia/reperfusion induced injury and oxidative change. Since the GNN conjugate markedly reduced elevated levels of tissue lipid peroxidation and attenuated renal dysfunction in rats subjected to renal ischemia/reperfusion, it might be possible to develop the GNN conjugate into a potential therapeutic agent against renal ischemia/reperfusion injury. PMID- 19555684 TI - Extracellular signal-regulated kinase 2 (ERK-2) mediated phosphorylation regulates nucleo-cytoplasmic shuttling and cell growth control of Ras-associated tumor suppressor protein, RASSF2. AB - Ras GTPase controls the normal cell growth through binding with an array of effector molecules, such as Raf and PI3-kinase in a GTP-dependent manner. RASSF2, a member of the Ras association domain family, is known to be involved in the suppression of cell growth and is frequently down-regulated in various tumor tissues by promoter hypermethylation. In the present study, we demonstrate that RASSF2 shuttles between nucleus and cytoplasm by a signal-mediated process and its export from the nucleus is sensitive to leptomycin B. Amino acids between 240 to 260 in the C-terminus of RASSF2 harbor a functional nuclear export signal (NES), which is necessary and sufficient for efficient export of RASSF2 from the nucleus. Substitution of conserved Ile254, Val257 and Leu259 within the minimal NES impaired RASSF2 export from the nucleus. In addition, wild type but not the nuclear export defective RASSF2 mutant interacts with export receptor, CRM-1 and exported from the nucleus. Surprisingly, we observed nucleolar localization for the nuclear export defective mutant suggesting the possibility that RASSF2 may localize in different cellular compartments transiently in a cell cycle dependent manner and the observed nuclear localization for wild type protein may be due to faster export kinetics from the nucleolus. Furthermore, our data suggest that RASSF2 is specifically phosphorylated by MAPK/ERK-2 and the inhibitors of MAPK pathway impair the phosphorylation and subsequently block the export of RASSF2 from the nucleus. These data clearly suggest that ERK-2 mediated phosphorylation plays an important role in regulating the nucleo-cytoplasmic shuttling of RASSF2. Interestingly, nuclear import defective mutant of RASSF2 failed to induce cell cycle arrest at G1/S phase and apoptosis suggesting that RASSF2 regulates cell growth in a nuclear localization dependent manner. Collectively, these data provided evidence for the first time that MAPK/ERK-2 mediated phosphorylation regulates nucleo-cytoplasmic transport and cell growth arrest activity of RASSF2. Taken together, the present study suggests that active transport between nucleus and cytoplasm may constitute an important regulatory mechanism for RASSF2 function. PMID- 19555686 TI - Protein ubiquitination in postsynaptic densities after hypoxia in rat neostriatum is blocked by hypothermia. AB - Synaptic dysfunction has been associated with neuronal cell death following hypoxia. The lack of knowledge on the mechanisms underlying this dysfunction prompted us to investigate the morphological changes in the postsynaptic densities (PSDs) induced by hypoxia. The results presented here demonstrate that PSDs of the rat neostriatum are highly modified and ubiquitinated 6 months after induction of hypoxia in a model of perinatal asphyxia. Using both two dimensional (2D) and three dimensional (3D) electron microscopic analyses of synapses stained with ethanolic phosphotungstic acid (E-PTA), we observed an increment of PSD thickness dependent on the duration and severity of the hypoxic insult. The PSDs showed clear signs of damage and intense staining for ubiquitin. These morphological and molecular changes were effectively blocked by hypothermia treatment, one of the most effective strategies for hypoxia-induced brain injury available today. Our data suggest that synaptic dysfunction following hypoxia may be caused by long-term misfolding and aggregation of proteins in the PSD. PMID- 19555685 TI - Morphine-induced nitric oxide production in isolated, iris-ciliary bodies. AB - Considerable evidence suggests that the nitric oxide (NO)/cGMP signaling pathway plays an integral role in opioid receptor-mediated responses in the cardiovascular and immune systems. Previous studies in our laboratory and others have shown that nitric oxide (NO) plays a role in morphine-induced reduction of intraocular pressure (IOP) and pupil diameter (PD) in the New Zealand white (NZW) rabbit. The present study is designed to determine the effect of morphine on NO production in the isolated, iris-ciliary body (ICB), site of aqueous humor production, as this effect could be associated with morphine-stimulated changes in aqueous humor dynamics and iris function. ICBs obtained from normal NZW rabbits were utilized in these experiments. In some experiments, ICB samples were treated with morphine (1, 10 and 100 microM) for 1 h and later examined for changes in NO levels using a NO detection kit. In other experiments, tissue samples were pretreated with naloxone (non-selective opioid receptor antagonist), L-NAME (non-selective NO-synthase inhibitor) or GSH (sulfhydryl reagent) for 30 min, followed by treatment with morphine (10 muM). Morphine caused a concentration-dependent increase in the release of NO from ICBs. Levels of NO detected in the incubation medium of ICB samples increased from 1.49 +/- 0.19 (control) to 8.81 +/- 2.20 microM/mg protein (morphine-treated; 100 microM). Morphine-stimulated release of NO was significantly inhibited in tissues pretreated with 10 microM naloxone, L-NAME, or GSH. Results obtained from this study suggest that morphine stimulates NO release from the ICB through a mechanism that involves activation of NO-releasing opioid receptors. These results support the in vivo effects of morphine demonstrated in previous studies. PMID- 19555687 TI - Identification of transplantable dopamine neuron precursors at different stages of midbrain neurogenesis. AB - Protocols used for generation of mesencephalic dopamine (mesDA) neurons from stem cells, or fetal brain tissue, invariably result in cell preparations that are highly mixed in composition, containing mesDA neuron precursors in various states of fate commitment and differentiation. For further optimisation and refinement of these procedures it is essential to determine the optimal stage of development and phenotypic characteristics of cells used for grafting. We have used fluorescence-activated cell sorting procedures to isolate mesDA precursors in defined stages of differentiation from mouse ventral mesencephalon (VM), at embryonic day 10.5 (E10.5), when the mesDA neuron domain consists of proliferative radial glia-like cells expressing the mesDA neuron determinant Lmx1a and the floorplate marker Corin, and at E12.5, when the VM has expanded to comprise a mixture of proliferative progenitors, neuroblasts and young neurons. The sorted cells were transplanted to the striatum of 6-hydroxydopamine-lesioned rats. Results show that the Lmx1a/Corin-expressing ventricular zone progenitors, which are the source of mesDA neurons in grafts from E10.5 VM, had lost this capacity at E12.5. At this later stage all transplantable mesDA precursors resided in the intermediate zone as postmitotic Nurr1-expressing neuroblasts. The more differentiated, TH-expressing cells survived sorting and transplantation poorly. We also provide evidence that, during early mesDA neurogenesis, the progenitors for nigral mesDA neurons segregate to lateral parts of the Lmx1a expressing domain and can be selectively isolated based on their level of Corin expression. These results have implications for current efforts to develop well characterized stem cell-derived mesDA progenitor cell preparations for cell therapy. PMID- 19555688 TI - Trichinella spiralis: Effect of thymus factor X on apoptosis and necrosis in mice. AB - The aim of the study was to determine the effect of thymus factor X (TFX-Jelfa) on the percentage of apoptotic and necrotic lymphocytes in the spleen, mesenteric lymph nodes, and muscle tissue of mice infected with 200 larvae of Trichinella spiralis. TFX was administered subcutaneously at a dose of 15mg/kg. On days 7, 14, 21, 28, 35, 42, and 60 after infection, apoptotic and necrotic cells were detected by flow cytometry after staining with the Annexin V-Fluos Staining Kit. TFX increased the percentage of apoptotic lymphocytes in the spleen, mesenteric lymph nodes, and muscle tissue of mice infected with T. spiralis. The effect of TFX on the percentage of necrotic lymphocytes was weaker and less clear. Parasite load was lower in infected mice treated with TFX than in the untreated control mice. The effect of TFX on the host immune response and the survival of parasite larvae was therefore probably affected by the extent of inflammatory infiltrates, and not by the percentage of lymphocytes undergoing apoptosis. PMID- 19555689 TI - MCC, a new interacting protein for Scrib, is required for cell migration in epithelial cells. AB - To further characterize the molecular events supporting the tumor suppressor activity of Scrib in mammals, we aim to identify new binding partners. We isolated MCC, a recently identified binding partner for beta-catenin, as a new interacting protein for Scrib. MCC interacts with both Scrib and the NHERF1/NHERF2/Ezrin complex in a PDZ-dependent manner. In T47D cells, MCC and Scrib proteins colocalize at the cell membrane and reduced expression of MCC results in impaired cell migration. By contrast to Scrib, MCC inhibits cell directed migration independently of Rac1, Cdc42 and PAK activation. Altogether, these results identify MCC as a potential scaffold protein regulating cell movement and able to bind Scrib, beta-catenin and NHERF1/2. PMID- 19555690 TI - Lanthanum ions inhibit the mammalian Sec61 complex in its channel dynamics and protein transport activity. AB - Previous electrophysiological experiments characterized the Sec61 complex, which provides the aqueous path for entry of newly-synthesized polypeptides into the mammalian endoplasmic reticulum, as a highly dynamic channel that, once activated by precursor proteins, fluctuates between main open states with mean conductances of 220 and 550pS. Millimolar concentrations of lanthanum ions simultaneously restricted the dynamics of the Sec61 channel and inhibited translocation of polypeptides. Molecular modeling indicates that lanthanum binding sites cluster at the putative lateral gate of the Sec61 complex and suggests that structural flexibility of the lateral gate is essential for channel and protein transport activities of the Sec61 complex. PMID- 19555691 TI - Expression profiling of the genes responding to zearalenone and its analogues using estrogen-responsive genes. AB - To compare gene expression profiles in response to estrogen or 17beta-estradiol (E(2)) and a mycotoxin, zearalenone (ZEA), and its analogues (collectively termed ZEA compounds), breast cancer MCF-7 cells were treated with 10 nM of E(2) or ZEA compounds including ZEA, alpha-zearalenol, beta-zearalenol, zearalanone, alpha zearalanol and beta-zearalanol. Expression profiles for 120 estrogen-responsive genes were subjected to cluster and statistical analyses using correlation coefficients or R-values. We found that all of the ZEA compounds stimulated the growth of MCF-7 cells, as much as E(2), and showed similar expression profiles to that of E(2) (R-values ranged from 0.82 to 0.96). The effect of ZEA compounds was likely mediated by estrogen-receptor-dependent Erk1/2-signaling. These results provide clues to understand the mechanism of their estrogen-like action. PMID- 19555692 TI - Immunoglobulin A: Fc(alpha)RI interactions induce neutrophil migration through release of leukotriene B4. AB - BACKGROUND & AIMS: Exacerbations of ulcerative colitis (UC) are dominated by massive neutrophil influx in the lamina propria with concomitant mucosal ulceration. The prevalent antibody in this area is immunoglobulin A (IgA). Interestingly, the IgA Fc receptor (Fc(alpha)RI) potently activates neutrophils. As such, we investigated whether IgA-Fc(alpha)RI interaction contributes to tissue damage in UC. METHODS: Response of neutrophils to bovine serum albumin-, IgG-, or IgA-coated beads and Escherichia coli was investigated with 3 dimensional culture systems, real-time video microscopy, and (fluorescence) microscopy. In vivo studies were performed using human Fc(alpha)RI transgenic mice or nontransgenic littermates. Microscopic slides of UC patients were stained for IgA, Fc(alpha)RI, and neutrophils. RESULTS: In vitro and in vivo cross linking of Fc(alpha)RI on neutrophils by serum IgA or uptake of IgA-coated E coli led to neutrophil migration. The responsible chemotactic factor was identified as leukotriene B4. Moreover, dimeric IgA (dIgA), which is produced in the lamina propria, but neither secretory IgA nor IgG, was equally capable of inducing neutrophil recruitment. We furthermore showed that Fc(alpha)RI(+)-neutrophils in the colon of UC patients had phagocytosed IgA-antigen complexes. CONCLUSIONS: Neutrophils are the first cells that arrive at inflammatory sites once pathogens have crossed the epithelial barrier. Fc(alpha)RI-dIgA interactions therefore may constitute an essential activation step to recruit more neutrophils, hereby eradicating impending infections. However, excessive IgA-antigen complexes can sustain a perpetuating inflammatory loop in UC, hereby seriously aggravating morbidity. Novel therapeutic strategies that block dIgA-Fc(alpha)RI interactions, and therefore diminish neutrophil migration and activation, may dampen the uncontrolled inflammatory processes in these patients. PMID- 19555693 TI - Salivary gland degeneration and vitellogenesis in the ixodid tick Amblyomma hebraeum: Surpassing a critical weight is the prerequisite and detachment from the host is the trigger. AB - The normal engorged body weight of female ixodid ticks (Acari: Ixodidae) is about 100x the unfed weight. Virgin female Amblyomma hebraeum normally do not feed beyond 10x the unfed weight. However, about 10-20% of a population of virgins will feed to perhaps 20x the unfed weight, but not much beyond that. In A. hebraeum, when females surpass about 10x the unfed weight, the following changes in physiology occur if they are removed from the host: (a) they will not reattach if given the opportunity, (b) their salivary glands (SGs) will undergo autolysis within 4 days if they are mated or 8 days if they are virgin, and (c) egg maturation and oviposition will occur in due course. Mated or virgin female ticks removed from the host below about 10x the unfed weight do not experience the latter changes (Kaufman, W.R., Lomas, L., 1996. 'Male Factors' in ticks: their role in feeding and egg development. Invertebrate Reproduction and Development 30, 191-198). In 1984 we named this transitional weight, the 'critical weight' (CW). Its absolute value is probably a species-specific characteristic (Kaufman, W.R., 2007. Gluttony and sex in female ixodid ticks: how do they compare to other blood-sucking arthropods? Journal of Insect Physiology 53, 264-273). Although mated females tend to engorge within a day of surpassing the CW, virgin females surpassing the CW can remain attached to the host for at least several weeks more. It is not known whether the physiological changes in the SGs and ovaries listed above occur in those large virgins that remain attached, although we suppose that this would be maladaptive. Instead, we hypothesize in this study that surpassing the CW is only a prerequisite for inducing these changes, and that detachment is the actual trigger. We support our hypothesis by demonstrating that large virgins, remaining attached to a host for 8 days, did not undergo SG degeneration nor complete egg maturation during the attachment period. Those changes occurred only within 8 days following detachment. So some type of sensory information associated with attachment to the host, and still undefined, inhibits expression of the physiological changes hitherto associated merely with surpassing the CW. PMID- 19555695 TI - Why do larval helminths avoid the gut of intermediate hosts? AB - In complex life cycles, larval helminths typically migrate from the gut to exploit the tissues of their intermediate hosts. Yet the definitive host's gut is overwhelmingly the most favoured site for adult helminths to release eggs. Vertebrate nematodes with one-host cycles commonly migrate to a site in the host away from the gut before returning to the gut for reproduction; those with complex cycles occupy sites exclusively in the intermediate host's tissues or body spaces, and may or may not show tissue migration before (typically) returning to the gut in the definitive host. We develop models to explain the patterns of exploitation of different host sites, and in particular why larval helminths avoid the intermediate host's gut, and adult helminths favour it. Our models include the survival costs of migration between sites, and maximise fitness (=expected lifetime number of eggs produced by a given helminth propagule) in seeking the optimal strategy (host gut versus host tissue exploitation) under different growth, mortality, transmission and reproductive rates in the gut and tissues (i.e. sites away from the gut). We consider the relative merits of the gut and tissues, and conclude that (i) growth rates are likely to be higher in the tissues, (ii) mortality rates possibly higher in the gut (despite the immunological inertness of the gut lumen), and (iii) that there are very high benefits to egg release in the gut. The models show that these growth and mortality relativities would account for the common life history pattern of avoidance of the intermediate host's gut because the tissues offer a higher growth rate/mortality rate ratio (discounted by the costs of migration), and make a number of testable predictions. Though nematode larvae in paratenic hosts usually migrate to the tissues, unlike larvae in intermediates, they sometimes remain in the gut, which is predicted since in paratenics mortality rate and migration costs alone determine the site to be exploited. PMID- 19555694 TI - Stressing the giant: a new approach to understanding dilated cardiomyopathy. PMID- 19555696 TI - Importance of the substrate-binding loop region of human monomeric carbonyl reductases in catalysis and coenzyme binding. AB - AIMS: Monomeric carbonyl reductase 1 (CBR1) and 3 (CBR3) are members of the short chain dehydrogenase/reductase superfamily, and metabolize endogenous and xenobiotic compounds using NADPH as a coenzyme. CBR3 exhibits a higher K(m) value toward NADPH and more limited carbonyl reductase activities than CBR1, although they are highly homologous to each other in amino acid sequence levels. In the present study, we investigated the origin of the different properties of the enzymes by analyses using several chimeric enzymes. MAIN METHODS: Harr-plot analysis of the amino acid sequences was conducted and as a result, two low identity regions between human CBR1 and CBR3 were found: these were designated as the N-terminal low-identity region (LirN) and the C-terminal low-identity region (LirC; the substrate-binding region). We genetically constructed chimeric enzymes while focusing on these regions. KEY FINDINGS: Chimeric CBR1 possessing LirN of CBR3 (CBR1LirN3) exhibited CBR1-like activities but a low coenzyme affinity probably due to a structural alteration in a micro domain, whereas chimeric CBR1 including LirC of CBR3 (CBR1LirC3) was enzymatically similar to CBR3. Furthermore, CBR3LirC1 was similar to CBR1 in both enzymatic activities and coenzyme binding. SIGNIFICANCE: These results suggested that LirC, i.e., the substrate-binding loop region, is the origin of the difference between human CBR1 and CBR3 in both catalytic and coenzyme-binding properties. PMID- 19555697 TI - Axon-reflex cutaneous vasodilatation is impaired in type 2 diabetic patients receiving chronic low-dose aspirin. AB - Low-dose aspirin is largely but non-homogeneously used in primary prevention of cardiovascular complication in type-2 diabetic patients. We hypothesised that low dose aspirin could interfere with the cutaneous neurovascular responses in type-2 diabetic patients. Galvanic current-induced vasodilatation (CIV) is an original non-noxious integrative model of neurovascular interaction and is impaired under low-dose aspirin in healthy subjects. Twenty type-2 diabetic patients (ten not receiving aspirin: D(-NA) and ten regularly receiving 21 drinks per week) the effect on total alcohol intake was maintained at five-year follow-up (net-change: -3.7; p=0.01). No significant effects were found in women on total alcohol intake. CONCLUSION: Multi-factorial lifestyle intervention, including low intensity alcohol intervention, improved long-term alcohol habits in a general population. PMID- 19555712 TI - Physical activity and prevalence of hypertension in a population-based sample of Brazilian adults and elderly. AB - OBJECTIVES: a) To determine the prevalence of hypertension and its associated factors and b) to establish the impact of past (i.e. adolescence) and current physical activity on hypertension. METHODS: A cross-sectional study of people aged 40 years or more living in the urban area of Pelotas - Brazil (2003). Hypertension was assessed by self-report of medical diagnosis. Individuals were considered active in adolescence if they reported regular physical activity practice between the ages 10 and 19 years. In adulthood, individuals who performed > or =150 min/week of leisure-time physical activities were considered active. Other variables studied included skin color, age, smoking, body mass index and socioeconomic status. RESULTS: 1696 individuals were interviewed (response rate >95%). Prevalence of hypertension was 34.4% (CI(95%) 32.1-36.7) and varied considerably among population subgroups. Current physical activity showed no association with hypertension. Physical activity in adolescence was associated with a decreased risk of adulthood hypertension in the crude analyses for women (p<0.001). However, after adjusting for potential confounders and mediators, this protection was no longer significant 1.20 (CI(95%) 0.93-1.55). CONCLUSIONS: Prevalence of hypertension is high among Brazilians and markedly different among population subgroups. There was no robust evidence of association between adolescence physical activity practice and hypertension in later life. PMID- 19555711 TI - Do vegetables and fruits reduce the risk of chronic obstructive pulmonary disease? A case-control study in Japan. AB - OBJECTIVE: To investigate the relationship between vegetable and fruit consumption and the risk of chronic obstructive pulmonary disease (COPD), a case control study was conducted in central Japan in 2006. METHODS: A total of 278 referred patients with COPD diagnosed within the past four years and 340 community-based controls undertook spirometric measurements of respiratory function. A structured questionnaire was administered face-to-face to obtain information on demographics, lifestyle and habitual food consumption. RESULTS: The mean vegetable and fruit intakes of cases (155.62 (SD 88.84) and 248.32 (SD 188.17) g/day) were significantly lower (p<0.01) than controls (199.14 (SD 121.41) and 304.09 (SD 253.72) g/day). A substantial reduction in COPD risk was found by increasing daily total vegetable intake, p for trend=0.037. The prevalence of breathlessness also decreased with vegetable consumption, the adjusted odds ratio being 0.49 (95% CI 0.27-0.88) for the highest versus lowest quartile of intake. However, the effects of fruit consumption were not significant. Among the nutrients contained in vegetables and fruits, vitamin A was particularly significant (p=0.008) with an estimated 52% reduction in COPD risk at the highest level of intake. CONCLUSION: The study provided evidence of an inverse association between vegetable consumption and the risk of COPD for Japanese adults. PMID- 19555713 TI - Quadrivalent Human Papillomavirus recombinant vaccine associated lipoatrophy. AB - Involutional lipoatrophy, a loss of subcutaneous fat, may be idiopathic, associated with inflammatory skin conditions, or trauma, and has also been reported following injections of medications including insulin, corticosteroids and penicillin. There have also been reports in association with Diptheria Pertussis Tetanus (DPT) vaccine. We report on two cases of lipoatrophy associated with the new Quadrivalent Human Papillomavirus (HPV) recombinant vaccine (Gardasil). PMID- 19555714 TI - Preparation of bacterial polysaccharide-protein conjugates: analytical and manufacturing challenges. AB - A conjugate can be a polysaccharide (PS) covalently attached to a protein, which provides T cell epitopes for a normally T cell independent antigen. To produce a conjugate vaccine, the purified PS must first be chemically modified to generate reactive groups that can link to the protein. Two commonly used methods for PS activation are periodate oxidation at vicinal hydroxyls and cyanylation of hydroxyls. The PS should be of known molecular size before and after activation. Low molecular weight impurities in the protein may result in inefficient conjugation. Two critical measures after conjugation and purification are the PS to protein ratio and the percent non-conjugated saccharide (free saccharide). Yield and conjugate stability are critical considerations. Typically, considerably less than 20% of the activated PS becomes conjugated. Yield can be improved using newer conjugation methods, whereby highly reactive groups are generated on both the PS and carrier protein with yields approaching 50%. Two major measures used to follow vaccine stability are changes in molecular size and percent free (unbound) PS. PMID- 19555715 TI - Cost of routine immunization of young children against rotavirus infection with Rotarix versus RotaTeq. AB - Using a probabilistic model of the clinical and economic burden of rotavirus gastroenteritis (RVGE), we estimated the expected impact of vaccinating a US birth cohort with Rotarix in lieu of RotaTeq. Assuming full vaccination of all children, use of Rotarix - rather than RotaTeq - was estimated to reduce the total number of RVGE events by 5% and associated costs by 8%. On an overall basis, Rotarix would reduce costs by $77.2 million (95% CI $71.5-$86.5). Similar reductions with Rotarix were estimated to occur under an assumption of incomplete immunization of children. PMID- 19555716 TI - Inflammatory mechanisms and treatment of obstructive airway diseases with neutrophilic bronchitis. AB - Obstructive airway diseases such as asthma and chronic obstructive pulmonary disease (COPD) are major global health issues. Although considered as distinct diseases, airway inflammation is a key underlying pathophysiological process in asthma, COPD and bronchiectasis. Persistent neutrophilic airway inflammation (neutrophilic bronchitis) occurs with innate immune activation and is a feature of each of these airway diseases. Little is known about the mechanisms leading to neutrophilic bronchitis and few treatments are effective in reducing neutrophil accumulation in the airways. There is a similar pattern of inflammatory mediator release and toll like receptor 2 expression in asthma, COPD and bronchiectasis. We propose the existence of an active amplification mechanism, an effector arm of the innate immune system, involving toll like receptor 2, operating in persistent neutrophilic bronchitis. Neutrophil persistence in the airways can occur through a number of mechanisms such as impaired apoptosis, efferocytosis and mucus hypersecretion, all of which are impaired in airways disease. Impairment of neutrophil clearance results in a reduced ability to respond to bacterial infection. Persistent activation of airway neutrophils may result in the persistent activation of the innate immune system resulting in further airway insult. Current therapies are limited for the treatment of neutrophilic bronchitis; possible treatments being investigated include theophylline, statins, antagonists of pro-inflammatory cytokines and macrolide antibiotics. Macrolides have shown great promise in their ability to reduce airway inflammation, and can reduce airway neutrophils, levels of CXCL8 and neutrophil proteases in the airways. Studies also show improvements in quality of life and exacerbation rates in airways diseases. PMID- 19555718 TI - Cellular models for the screening and development of anti-hepatitis C virus agents. AB - Investigations on the biology of hepatitis C virus (HCV) have been hampered by the lack of small animal models. Efforts have therefore been directed to designing practical and robust cellular models of human origin able to support HCV replication and production in a reproducible, reliable and consistent manner. Many different models based on different forms of virions and hepatoma or other cell types have been described including virus-like particles, pseudotyped particles, subgenomic and full length replicons, virion productive replicons, immortalised hepatocytes, fetal and adult primary human hepatocytes. This review focuses on these different cellular models, their advantages and disadvantages at the biological and experimental levels, and their respective use for evaluating the effect of antiviral molecules on different steps of HCV biology including virus entry, replication, particles generation and excretion, as well as on the modulation by the virus of the host cell response to infection. PMID- 19555717 TI - Cooperative properties of cytochromes P450. AB - Cytochromes P450 form a large and important class of heme monooxygenases with a broad spectrum of substrates and corresponding functions, from steroid hormone biosynthesis to the metabolism of xenobiotics. Despite decades of study, the molecular mechanisms responsible for the complex non-Michaelis behavior observed with many members of this superfamily during metabolism, often termed 'cooperativity', remain to be fully elucidated. Although there is evidence that oligomerization may play an important role in defining the observed cooperativity, some monomeric cytochromes P450, particularly those involved in xenobiotic metabolism, also display this behavior due to their ability to simultaneously bind several substrate molecules. As a result, formation of distinct enzyme-substrate complexes with different stoichiometry and functional properties can give rise to homotropic and heterotropic cooperative behavior. This review aims to summarize the current understanding of cooperativity in cytochromes P450, with a focus on the nature of cooperative effects in monomeric enzymes. PMID- 19555720 TI - TLR5 ligation by flagellin converts tolerogenic dendritic cells into activating antigen-presenting cells that preferentially induce T-helper 1 responses. AB - Dendritic cells (DCs) differentiated in the presence of IL-10 preferentially induce regulatory T-cells and tolerance. Whether the tolerogenic properties displayed by these DCs (Tol-DCs) can be overcome has not been fully explored. Here we show for the first time that Tol-DCs express higher levels of TLR5 mRNA, but not TLR4 or TLR9 mRNA relative to DCs differentiated with GM-CSF and IL-4 (BM DCs). In response to flagellin, a natural TLR-5 ligand, Tol-DCs produced IL-12 but not IL-10. Unlike Tol-DCs stimulated with LPS, which produce high levels of IL-10 and fail to generate a cognate inflammatory response in CD4(+) T-cells, flagellin-stimulated Tol-DCs promoted the differentiation of CD4(+) T cells with a T-helper 1 phenotype. The divergent T-cell outcomes induced by Tol-DCs in response to different TLR-ligands highlights not only their plasticity, but also points to TLR5 ligation as a potential strategy to overcome tolerance in environments that are otherwise conducive to immune unresponsiveness. PMID- 19555721 TI - The effect of pre-treatment and sonication of centrifugal ultrafiltration devices on virus recovery. AB - Enteric viruses such as norovirus (NV) and hepatitis A (HAV) are responsible for a large proportion of food and water-borne illnesses. Most human pathogenic enteric viruses cannot be cultured so they must be detected by molecular techniques. Male specific (F(+)) RNA coliphages, a potential surrogate for human enteric viruses, can be detected by culture and molecular assays. Numbers of viruses and F-RNA coliphages in contaminated food or water may be too low for direct detection. Ultrafiltration is a general concentration method for all virus types but there is little information on the recovery efficiency of F-RNA coliphages and enteric viruses. The recovery of F-RNA coliphage MS2 was only 25% by plaque assay in initial trials. The objective was to optimize the recovery of concentrated MS2 from Microsep 100K ultrafiltration devices. The mean recovery of MS2 increased significantly to 85% by plaque assay and 65% by real-time RT-PCR when ultrafiltration devices were treated with 1% BSA before concentration and then ultrasonicated after concentration. The method was validated with MS2, HAV, NV and feline calicivirus (FCV) in water and spinach eluate. The recovery of MS2, HAV and NV was significantly higher from concentrates obtained from water with treated devices than untreated devices but not significantly different for FCV or from spinach eluate. To our knowledge, this is the first study to use ultrasonication as a post-treatment step to increase recovery of viruses from ultrafiltration devices. PMID- 19555722 TI - Comparative analysis of in vitro processivity of HIV-1 reverse transcriptases containing mutations 65R, 74V, 184V and 65R+74V. AB - While HIV-1 reverse transcriptase (RT) mutations of M to V at position 184 are commonly observed in the clinic, the double mutation of 65R+74V is rarely seen. It has been demonstrated that rapid R-->K reversion occurs at RT codon 65 during replication of HIV-1 in human peripheral blood mononuclear cells containing 65R+74V mutations and that processivity of the RT is reduced relative to wild type. However, clinical studies show that M184V can be detected after several months of therapy interruption, suggesting more effective processivity. Herein, the in vitro RT processivity of genetically engineered M184V and double mutant 65R+74V was compared. Virion-associated RTs of WT pNL4-3, K65R, L74V, M184V and 65R+74V were used to perform RT processivity assays in the presence of trap, poly(rC)-oligo(dG). Both RTs with 184V and 65R+74V mutations exhibited similar processivity when compared with each other and a significantly decreased processivity as compared to WT RT. Both mutant RTs synthesized shorter cDNA molecules (37-42 nt) relative to WT RT, which made longer (65-70 nt) cDNA molecules. Since these surprising biochemical results cannot explain the clinical phenotype, a hypothesis is presented to explain the discrepancy and suggest new approaches for future testing. PMID- 19555724 TI - Positive reinforcing effects of substance P in the rat central nucleus of amygdala. AB - Substance P (SP) can have positive reinforcing or aversive properties, depending on the dose used and the site of action in the brain. Experimental findings suggest that the amygdala is involved in reward-related processes. The presence of SP-immunoreactive fibers and cell bodies has been shown in the central nucleus (ACE) and neurokinin (NK)-1 and NK-3 receptors also could be found there. The rewarding or aversive effects of SP in the ACE were tested in conditioned place preference paradigm. 10 ng SP microinjections had positive reinforcing properties, while 100 ng SP had no effect. Prior treatment with NK-1 receptor antagonist could block the rewarding effects of SP, while the antagonist on its own did not influence place preference. Our results show that SP and NK-1 receptors play important roles in amygdaloid rewarding-reinforcing mechanisms. PMID- 19555725 TI - Evaluation of a highly discriminating multiplex multi-locus variable-number of tandem-repeats (MLVA) analysis for Vibrio cholerae. AB - Vibrio cholerae is the etiological agent of cholera and may be used in bioterror actions due to the easiness of its dissemination, and the public fear for acquiring the cholera disease. A simple and highly discriminating method for connecting clinical and environmental isolates of V. cholerae is needed in microbial forensics. Twelve different loci containing variable numbers of tandem repeats (VNTRs) were evaluated in which six loci were polymorphic. Two multiplex reactions containing PCR primers targeting these six VNTRs resulted in successful DNA amplification of 142 various environmental and clinical V. cholerae isolates. The genetic distribution inside the V. cholerae strain collection was used to evaluate the discriminating power (Simpsons Diversity Index=0.99) of this new MLVA analysis, showing that the assay have a potential to differentiate between various strains, but also to identify those isolates which are collected from a common V. cholerae outbreak. This work has established a rapid and highly discriminating MLVA assay useful for track back analyses and/or forensic studies of V. cholerae infections. PMID- 19555726 TI - An optimized protocol for the production of interdelta markers in Saccharomyces cerevisiae by using capillary electrophoresis. AB - The amplification of genomic sequence blocks flanked by delta elements of retrotransposon origin has proved to be a very convenient method for molecular characterization of Saccharomyces cerevisiae strains. Fluorescent automated capillary electrophoresis (CE) was used to detect interdelta marker (IDM) patterns in S. cerevisiae, using the ABI Prism 3130 Genetic Analyzer. Main experimental parameters were studied and the optimal conditions for IDM amplification and samples run on the CE apparatus were determined. Fingerprints from fluorescent-labelled IDM produced using CE with the same sample analyzed by agarose electrophoresis (AE) were compared. The CE analysis was able to distinguish 43 different IDM profiles among 45 S. cerevisiae isolates with a discriminating capacity of 99.8%, whereas the AE analysis of the same samples allowed the identification of 27 different patterns (discriminatory power equal to 96%). Detection of fluorescent IDM was fast and reliable, and it facilitated data comparison. For the first time in our knowledge, the fluorescent CE proved to be well suited for IDM fingerprinting. Moreover, it could be routinely applied for the molecular differentiation of S. cerevisiae strains. PMID- 19555719 TI - Bipolar disorder and mechanisms of action of mood stabilizers. AB - Bipolar disorder (BD) is a major medical and social burden, whose cause, pathophysiology and treatment are not agreed on. It is characterized by recurrent periods of mania and depression (Bipolar I) or of hypomania and depression (Bipolar II). Its inheritance is polygenic, with evidence of a neurotransmission imbalance and disease progression. Patients often take multiple agents concurrently, with incomplete therapeutic success, particularly with regard to depression. Suicide is common. Of the hypotheses regarding the action of mood stabilizers in BD, the "arachidonic acid (AA) cascade" hypothesis is presented in detail in this review. It is based on evidence that chronic administration of lithium, carbamazepine, sodium valproate, or lamotrigine to rats downregulated AA turnover in brain phospholipids, formation of prostaglandin E(2), and/or expression of AA cascade enzymes, including cytosolic phospholipase A(2), cyclooxygenase-2 and/or acyl-CoA synthetase. The changes were selective for AA, since brain docosahexaenoic or palmitic acid metabolism, when measured, was unaffected, and topiramate, ineffective in BD, did not modify the rat brain AA cascade. Downregulation of the cascade by the mood stabilizers corresponded to inhibition of AA neurotransmission via dopaminergic D(2)-like and glutamatergic NMDA receptors. Unlike the mood stabilizers, antidepressants that increase switching of bipolar depression to mania upregulated the rat brain AA cascade. These observations suggest that the brain AA cascade is a common target of mood stabilizers, and that bipolar symptoms, particularly mania, are associated with an upregulated cascade and excess AA signaling via D(2)-like and NMDA receptors. This review presents ways to test these suggestions. PMID- 19555723 TI - Effects of stress, corticosterone, and epinephrine administration on learning in place and response tasks. AB - These experiments examined the effects of prior stress, corticosterone, or epinephrine on learning in mazes that can be solved efficiently using either place or response strategies. In a repeated stress condition, rats received restraint stress for 6h/day for 21 days, ending 24h before food-motivated maze training. In two single stress conditions, rats received a 1-h episode of restraint stress ending 30 min or 24h prior to training. Single stress ending 30 min prior to training resulted in a significant interaction of stress and learning on the two tasks, with significant enhancement of learning in the response task and non-significant impairment in the place task. Neither acute nor chronic stress significantly altered learning in either task when the stress ended 24h before training. Thus, the anterograde effects of stress on maze learning ended within a single day. Two stress-related hormones, corticosterone and epinephrine, were tested for effects on learning parallel to those of acute stress. When administered 30 min prior to training, a corticosterone dose (40 mg/kg) that enhanced memory on a spontaneous alternation task did not significantly enhance or impair learning in either task. Two doses of epinephrine that modulate memory in other settings were used to test the effects of epinephrine on learning. Pre-training injections of 0.03 mg/kg epinephrine impaired place learning, while 0.1mg/kg epinephrine impaired response learning. The epinephrine results mimicked those seen with acute stress on the place task, but were opposite those seen after acute stress on the response task. Thus, corticosterone does not appear to be a major factor mediating the effects of acute stress on place and response learning and epinephrine is, at most, a partial contributor to these effects. PMID- 19555727 TI - Comparison of multiplex PCR, enzyme immunoassay and cell culture methods for the detection of enterotoxinogenic Bacillus cereus. AB - Fast and reliable methods are needed for the detection of pathogenic Bacillus cereus which should provide consistent results. Therefore, we tested a panel of 176 strains, including B. cereus strains, B. cereus group strains and other Bacillus spp. with polymerase chain reaction, immunoassays and cytotoxicity tests and assessed the consistency of the results. A screening multiplex PCR for the detection of hbl, nhe, ces and cytK1 as well as two multiplex PCRs for the differentiation of Hbl genes (hblC, hblD, hblA) and Nhe genes (nheA, nheB, nheC) was applied. All PCRs included an internal amplification control. Component specific antibody based immunoassays were used for the detection of the three components of Hbl and Nhe and the overall cytotoxicity to Vero cells and HEp-2 cells was checked. An overall excellent correlation was obtained for the results of the three, methodically independent assays and no false-negative PCR results were seen for any of the strains tested positive in immunoassays and cytotoxicity tests. The three multiplex PCRs proved to be a facile method for the identification of enterotoxinogenic B. cereus isolates. PMID- 19555728 TI - Saturation-mutagenesis in two positions distant from active site of a Klebsiella pneumoniae glycerol dehydratase identifies some highly active mutants. AB - Synthesis of 1,3-propanediol (1,3-PD) from glycerol through the biotransformation process requires two steps, catalyzed by glycerol dehydratase (GDHt) and 1,3-PD oxidoreductase. GDHt is the rate-limiting enzyme in this process. All recombinant microorganisms for production of 1,3-PD so far utilized the natural genes that may not have been optimized. Two positions, which are 19.3A and 29.6A away from the active site in GDHt from Klebsiella pneumoniae, were subjected to saturation mutagenesis and 38 mutants were characterized. The catalytic activity of a mutant in beta-subunit (beta-Q42F, 29.6A from the active site) was 8.3-fold higher than the wild type, and the enzyme efficiency of other two mutants beta-Q42L and beta Q42S for substrate glycerol was 336-fold and 80-fold higher than that for 1,2 propanediol. This investigation supplied further evidence that distant mutations could be a good source of diversity and therefore, made a contribution to the toolbox of industrial enzyme improvement. PMID- 19555729 TI - Alternation between foods within a meal. Influence on satiation and consumption in humans. AB - Food habituation/dishabituation has been observed in non-human primates in neurophysiological investigations of feeding, and in humans, through salivation or hedonic responses to food. The objective of the study was to evaluate in humans the effect of disruption of habituation by alternation between foods in a meal. Sixteen volunteers (8 males, 8 females; age: 21+/-1 yr; BMI: 21.5+/-0.5 kgm(-2)) ate a two-course meal [meatballs (M) and fries (F), then vanilla cream (C) and brownies (B)] during three randomized sessions. Sessions differed by the alternation of these foods: No-Repetition session with M-F-C-B; Single-Repetition session with F-M-F-B-C-B; Multiple-Repetition session with M-F-M-F-M-F-C-B-C-B-C B. Final intakes of F and B were ad libitum. Quantities consumed (g, kJ) and ratings of hunger, pleasantness and desire to eat each food were evaluated. Compared to the No-Repetition session, subjects ate 18% more fries and 16% more brownies in the Single-Repetition, and 13% more fries but 20% less brownies in the Multiple-Repetition session. Pleasantness for the food decreased from before to after intake for both fries and brownies with no significant difference between the sessions. It therefore appears that moderate alternation between foods at lunch increases intake, but multiple alternations of foods at the end of the meal may decrease consumption. These differences in intakes could result from differences in sensory-specific satiety via disruption of habituation. PMID- 19555730 TI - Alloantibodies in heart transplantation. AB - The presence of complement fixing anti-human leukocyte antigen (HLA) antibodies in the circulation of organ transplant recipients may result in heart allograft rejection. Here, we assessed the clinical impact of pre- and post-transplantation allosensitization on long-term survival of heart allografts. Sequential samples of sera from heart allograft recipients were screened pretransplantation for panel reactive antibodies using the complement-dependent cytotoxicity test. Patients were monitored post-transplantation for donor specific anti-HLA class I and class II antibodies. Kaplan-Meier graft survival plots were generated to analyze the effect of anti-HLA antibodies on transplantation outcomes. Statistical analysis showed that the post-transplantation development of alloantibodies was a significant risk factor that was associated with low long term survival rates; in contrast, recipients' gender, age, previous transplantations, and degree of HLA matching with the donor had no effect on long term survival. The presence in pretransplantation sera of antibodies against more than 10% of the HLA reference panel (PRA >10%) was associated with AMR and with a relatively lower rate of graft survival after 1 year but did not affect 10-year survival. The present data underline the importance of monitoring the development of anti-HLA antibodies as a tool for early diagnosis and treatment of AMR. PMID- 19555731 TI - Paliperidone for treatment of obsessive compulsive resistant symptoms in schizophrenia: a case report. PMID- 19555732 TI - Antihyperlipidemic effect of Garlip, a polyherbal formulation in streptozotocin induced diabetic rats. AB - This study was undertaken to investigate the effect of Garlip, a polyherbal drug composed of aqueous extract of six medicinal plants on blood glucose, plasma insulin, tissue lipid profile, and lipidperoxidation in streptozotocin induced diabetic rats. Aqueous extract of Garlip a, polyherbal drug was administered orally (200 mg/kg body weight) for 30 days. The different doses of Garlip on blood glucose and plasma insulin in diabetic rats were studied and the levels of lipid peroxides (TBARS and Hydroperoxide) and tissue lipids (cholesterol, triglyceride, phospholipids and free fatty acids) were also estimated in streptozotocin induced diabetic rats. The effects were compared with tolbutamide. Treatment with Garlip and tolbutamide resulted in a significant reduction of blood glucose and increase in plasma insulin. Garlip also resulted in a significant decrease in tissue lipids and lipid peroxide formation. The effect produced by Garlip was comparable with that of tolbutamide. The decreased lipid peroxides and tissue lipids clearly showed the antihyperlipidemic and antiperoxidative effect of Garlip apart from its antidiabetic effect. PMID- 19555733 TI - The garlic ingredient diallyl sulfide induces Ca(2+) mobilization in Madin-Darby canine kidney cells. AB - Diallyl sulfide (DAS), one of the major organosulfur compounds (OSCs) of garlic, is recognized as a group of potential chemoproventive compounds. In this study, we examines the early signaling effects of DAS on renal cells loaded with Ca(2+) sensitive dye fura-2. It was found that DAS caused an immediate and sustained rise of [Ca(2+)](i) in a concentration-dependent manner (EC(50)=2.32 mM). DAS also induced a [Ca(2+)](i) elevation when extracellular Ca(2+) was removed, but the magnitude was reduced by 45%. Depletion of intracellular Ca(2+) stores with CCCP, a mitochondrial uncoupler, did not affect DAS's effect. In Ca(2+)-free medium, the DAS-induced [Ca(2+)](i) rise was abolished by depleting stored Ca(2+) with thapsigargin (an endoplasmic reticulum Ca(2+) pump inhibitor). DAS-caused [Ca(2+)](i) rise in Ca(2+)-containing medium was not affected by modulation of protein kinase C activity. The DAS-induced Ca(2+) influx was blocked by nicardipine. U73122, an inhibitor of phospholipase C, abolished ATP (but not DAS) induced [Ca(2+)](i) rise. Additionally, pretreatment with DAS for 24 h decreased cell viability in a concentration-dependent manner. Furthermore, DAS-induced cell death involved apoptotic events. These findings suggest that diallyl sulfide induced a significant rise in [Ca(2+)](i) in MDCK renal tubular cells by stimulating both extracellular Ca(2+) influx and thapsigargin-sensitive intracellular Ca(2+) release via as yet unidentified mechanisms. PMID- 19555734 TI - Meeting report: second international meeting on quadruplex DNA. AB - A two and a half day meeting on G-quadruplexes was held in Louisville, KY, USA (April 18-21, 2009). A specific goal of this conference was to promote discussion on the biology of G-quadruplexes. In practice this was represented in four main ways, namely in biophysics, bio/nanotechnology, therapeutics, and what might be termed "intrinsic biology". Research into the basic biophysical and structural properties of G-quadruplexes continues to be important for understanding biology, and for optimizing aptamers for therapeutic and bio/technological purposes. The meeting comprised two Keynote lectures, twenty-three invited talks, and forty-two posters covering various aspects of these topics using a wide variety of technologies. PMID- 19555735 TI - Hepcidin, the iron watcher. AB - Hepcidin, a peptide hormone produced by the liver, constitutes the master regulator of iron homeostasis in mammals allowing iron adaptation according to the body iron needs. In recent years there has been important breakthrough in our knowledge of hepcidin regulation that has also implications for understanding the physiopathology of human iron disorders. Different aspects of hepcidin regulation will be considered in this review, including regulation by the iron status and the BMP6/HJV/SMAD pathway. Hepcidin dysregulation in iron disorders will be also discussed. Although much can already be accomplished for treating iron disorders using the knowledge that has currently been developed, additional issues will be challenging for the coming years. PMID- 19555736 TI - Analysis of stochasticity in promoter activation by using a dual-fluorescence reporter system. AB - Stochastic dynamics of promoter activity in bacterial cells were studied by using a dual-fluorescence reporter system of protein expression. The dual-fluorescence reporter system enabled us to calculate the amplitude of intrinsic noise generated during transcription and translation. By fitting the experimental data to a simple stochastic model of protein expression, we could estimate parameters representing the stochastic transition between the active and inactive states of a promoter. Using the system, we analyzed the stochastic dynamics of promoter activation of genes in the lysine biosynthesis pathway in Escherichia coli. We found that the promoter of lysA has a significantly slower transition rate between active and inactive states than other promoters in the lysine biosynthesis pathway. The infrequent switching between active and inactive states can be a dominant source of noise in lysA expression. Analysis using the dual fluorescence reporter system provided a better understanding of stochastic dynamics in promoter activation. PMID- 19555737 TI - Temporal changes in mouse brain fatty acid amide hydrolase activity. AB - Fatty acid amide hydrolase (FAAH) activity is known to mediate the tone of endogenous fatty acid amides including the endocannabinoid anandamide. FAAH is a potential therapeutic target because genetic or pharmacological ablation of FAAH promotes analgesia and anxiolytic effects without disrupting motor coordination. Little is known about the endogenous temporal fluctuations of brain FAAH activity. This is the first comprehensive study examining temporal fluctuations in mouse brain FAAH activity. Regional mouse brain homogenates were generated at the midpoint of the light ("noon") and dark ("midnight") cycles. While immunoblots revealed no significant changes (P>0.05) in regional activity between these two time points, in vitro activity assays detected a subtle 10% reduction (P<0.05) in cerebellar FAAH activity at midnight. A novel ex vivo autoradiography technique permitted the study of 11 different brain regions, many of which cannot be studied using traditional in vitro methods. The cerebellum and the periaqueductal gray both exhibited significant (P<0.05) reductions in regional FAAH activity in "midnight" brains. These data confirm the need to account for temporal changes in FAAH activity when therapeutically targeting FAAH. PMID- 19555738 TI - Phase coupling between rhythmic slow activity and gamma characterizes mesiotemporal rapid-eye-movement sleep in humans. AB - In the human sleep literature there is much controversy regarding the existence and the characteristics of hippocampal rhythmic slow activity (RSA). Generally the human RSA is believed to occur in short bursts of theta activity. An earlier study, however, reported mesiotemporal RSA during rapid-eye-movement (REM) sleep that instead of theta fell in the delta frequency band. We conjectured that if this RSA activity is indeed a human analogue of the animal hippocampal theta then characteristics associated with the animal theta should also be reflected in the human recordings. Here our aim was to examine possible phase coupling between mesiotemporal RSA and gamma activity during REM sleep. The study relied on nine epilepsy surgery candidates implanted with foramen ovale electrodes. Positive half-waves of the 1.5-3 Hz RSA were identified by an automatic algorithm during REM sleep. High-frequency activity was assessed for 11 consecutive 20 Hz-wide frequency bands between 20 and 240 Hz. Increase in high frequency activity was phase coupled with RSA in most frequency bands and patients. Such a phase coupling closely resembles that seen between theta and gamma in rodents. We consider this commonality to be an additional reason for regarding delta rather than theta as the human analogue of RSA in animals. PMID- 19555739 TI - Tau hyperphosphorylation affects Smad 2/3 translocation. AB - Transforming growth factors beta (TGFbeta) regulate multiple biological activities. TGFbeta activation of the Smad pathway results in activation of genes encoding extracellular matrix molecules, proteases, protease activators and protease inhibitors. In Alzheimer's disease (AD), TGFbeta protein and mRNA levels are raised, which would be expected to be neuroprotective. However, recent observations suggest that TGFbeta-Smad signalling is disrupted by the hyperphosphorylation of tau, the primary component of neurofibrillary tangles: phosphorylated Smad2/3 (pSmad 2/3) co-localises with phosphorylated tau in the neuronal cytoplasm and levels are reduced in the nucleus. We have investigated whether in vitro induction of tau hyperphosphorylation influences pSmad 2/3 localisation in rat primary cortical cells. Treatment with okadaic acid, a protein phosphatase 1 and 2A inhibitor caused hyperphosphorylation of tau at epitopes hyperphosphorylated in AD and disrupted pSmad 2/3 translocation into the nucleus. The disruptive effect of tau phosphorylation on pSmad 2/3 translocation was confirmed by treatment of primary cortical cells with synthetic oligomeric A beta(1-42), a more physiologically relevant model of AD. Our findings suggest that despite the increased level of TGFbeta in AD, the TGFbeta-Smad signalling pathway is impeded within neurones due to sequestration of pSmad 2/3 by hyperphosphorylated tau. This may compromise neuroprotective actions of TGFbeta and contribute to neurodegeneration in AD. PMID- 19555740 TI - Sustained depolarization decreases calcium/calmodulin-dependent protein kinase II activity and gene expression in dopamine neurons. AB - Altered gene expression mediated by calcium/calmodulin-dependent protein kinase II (CaMKII) and other intracellular signaling molecules plays an important role in activity-dependent neuroplasticity. We discovered that sustained depolarization induced by KCl, a commonly used paradigm for studying activity dependent gene expression, surprisingly caused a decrease in CaMKII activity in rat mesencephalic dopamine neurons. This decrease in CaMKII activity, after 2 days of depolarization, occurred in the presence of a continued elevation in intracellular calcium concentration. An increase in calyculin-sensitive phosphatase activity was at least partly responsible for the decrease in CaMKII activity. Phosphatase assays revealed that activity but not the abundance of protein phosphatase-2A was increased by sustained depolarization. Decreased CaMKII activity was accompanied by a selective decrease in dopamine transporter (DAT) mRNA, while tyrosine hydroxylase and actin mRNA abundance was unaltered. On the other hand, brain-derived neurotrophic factor (BDNF) mRNA abundance was increased by sustained depolarization, further demonstrating the specificity of changes. Depolarization also caused a significant decrease in DAT protein abundance and DAT-mediated uptake. Taken together, these data illustrate a novel signaling paradigm in which the activity of protein phosphatase-2A is associated with CaMKII activity and gene expression. PMID- 19555741 TI - Activation of purinergic receptors induces proliferation and neuronal differentiation in Swiss Webster mouse olfactory epithelium. AB - In the central nervous system (CNS), adenosine 5'-triphosphate (ATP) induces the synthesis and release of neurotrophic factors, cell proliferation, and differentiation. The olfactory system is one site where multipotent progenitor cells continue to proliferate and differentiate into neurons throughout life. We tested the hypothesis that ATP initiates proliferation in olfactory epithelium by measuring 5-bromo-2-deoxyuridine incorporation. Adult mice were pre-treated intraperitoneally (i.p.) or intranasally with saline or purinergic receptor antagonists (pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate+suramin) 30 min prior to nasal instillation of ATP, uridine 5'-triphosphate (UTP), adenosine 5' (3-thiotriphosphate) (ATP gamma S) or saline (0 h). Mice received three injections of 5-bromo-2-deoxyuridine between 42 and 46 h, and were sacrificed at 2, 9 or 16 days post-ATP instillation. ATP, UTP or ATP gamma S significantly increased 5-bromo-2-deoxyuridine incorporation compared to intranasal saline controls in groups pre-treated with saline. Saline, ATP, UTP or ATP gamma S instillation did not significantly increase 5-bromo-2-deoxyuridine incorporation in groups pre-treated with purinergic receptor antagonists. Similar results were observed in neonates and in a cultured slice preparation. Intranasal instillation of ATP also increased the protein levels of proliferating cell nuclear antigen in adults. Pre-treatment with purinergic receptor antagonists inhibited the ATP induced increase in proliferating cell nuclear antigen. In adults, a subset of the cells that incorporated 5-bromo-2-deoxyuridine was immunoreactive to neuronal markers mammalian achaete-schute homolog 1, growth-associated protein 43, and olfactory marker protein at 2, 9, and 16 days, respectively. Collectively, these data indicate that purinergic receptor activation induces proliferation and neuronal differentiation in the mouse olfactory epithelium. We propose that extracellular ATP released upon injury could induce proliferation and promote the neuroregeneration of the olfactory epithelium. PMID- 19555743 TI - The presenilin-1 familial Alzheimer's disease mutation P117L decreases neuronal differentiation of embryonic murine neural progenitor cells. AB - The presenilin-1 gene is mutated in early-onset familial Alzheimer's disease. The mutation Pro117Leu is implicated in a very severe form of the disease, with an onset of less than 30 years. The consequences of this mutation on neurogenesis in the hippocampus of adult transgenic mice have already been studied in situ. The survival of neural progenitor cells was impaired resulting in decreased neurogenesis in the dentate gyrus. Our intention was to verify if similar alterations could occur in vitro in progenitor cells from the murine ganglionic eminences isolated from embryos of this same transgenic mouse model. These cells were grown in culture as neurospheres and after differentiation the percentage of neurons generated as well as their morphology were analysed. The mutation results in a significant decrease in neurogenesis compared to the wild type mice and the neurons grow longer and more ramified neurites. A shift of differentiation towards gliogenesis was observed that could explain decreased neurogenesis despite increased proliferation of neural precursors in transgenic neurospheres. A diminished survival of the newly generated mutant neurons is also proposed. Our data raise the possibility that these alterations in embryonic development might contribute to increase the severity of the Alzheimer's disease phenotype later in adulthood. PMID- 19555744 TI - Concurrent performance in a three-alternative choice situation: response allocation in a Rock/Paper/Scissors game. AB - Adult human subjects engaged in a simulated Rock/Paper/Scissors game against a computer opponent. The computer opponent's responses were determined by programmed probabilities that differed across 10 blocks of 100 trials each. Response allocation in Experiment 1 was well described by a modified version of the generalized matching equation, with undermatching observed in all subjects. To assess the effects of instructions on response allocation, accurate probability-related information on how the computer was programmed to respond was provided to subjects in Experiment 2. Five of 6 subjects played the counter response of the computer's dominant programmed response near-exclusively (e.g., subjects played paper almost exclusively if the probability of rock was high), resulting in minor overmatching, and higher reinforcement rates relative to Experiment 1. On the whole, the study shows that the generalized matching law provides a good description of complex human choice in a gaming context, and illustrates a promising set of laboratory methods and analytic techniques that capture important features of human choice outside the laboratory. PMID- 19555745 TI - Coyotes (Canis latrans) and the matching law. AB - Environmental change is accelerating due to anthropogenic influence. Species that have greater behavioral flexibility may be better adapted to exploit new or constantly changing habitats. There are few mammals and even fewer carnivores that better illustrate widespread adaptability and behavioral flexibility in the wake of human disturbance than coyotes (Canis latrans). Yet how such predators successfully track resources, enabling them to survive and extend their range in stochastic environments remains unknown. We tested eight wild-born, captive coyotes individually on an operant two-choice test using concurrent variable interval (VI) schedules. We held the overall rate of reinforcement constant but manipulated the ratio of reinforcement available from the two choices. We analyzed sensitivity of coyotes' tracking of resource change by fitting the generalized matching equation to the data. Results showed all coyotes efficiently tracked changes in reinforcement ratios within the first few sessions of each new condition and matched their relative rate of foraging time to relative rate of resources. We suggest the matching paradigm provides a methodology to explore coyote foraging strategies, and a potential framework to compare behavioral flexibility across species, by measuring the ability to track resource change under variable resource conditions. PMID- 19555742 TI - Functional outcome is impaired following traumatic brain injury in aging Nogo-A/B deficient mice. AB - Increasing age is associated with a poor prognosis following traumatic brain injury (TBI). CNS axons may recover poorly following TBI due to expression of myelin-derived inhibitors to axonal outgrowth such as Nogo-A. To study the role of Nogo-A/B in the pathophysiological response of the elderly to TBI, 1-year-old mice deficient in Nogo-A/B (Nogo-A/B homozygous(-/-) mice), Nogo-A/B heterozygous(-/+) mice, and age-matched wild-type (WT) littermate controls were subjected to a controlled cortical impact (CCI) TBI. Sham-injured WT mice (7 months old) and 12 month old naive Nogo-A/B(-/-) and Nogo-A/B(-/+) served as controls. Neurological motor function was evaluated up to 3 weeks, and cognitive function, hemispheric tissue loss, myelin staining and hippocampal beta-amyloid (A beta) immunohistochemistry were evaluated at 4 weeks post-injury. In WT littermates, TBI significantly impaired learning ability at 4 weeks and neurological motor function up to 2 weeks post-injury and caused a significant loss of hemispheric tissue. Following TBI, Nogo-A/B(-/-) mice showed significantly less recovery from neurological motor and cognitive deficits compared to brain-injured WT mice. Naive Nogo-A/B(-/-) and Nogo-A/B(-/+) mice quickly learned the MWM task in contrast to brain-injured Nogo-A/B(-/-) mice who failed to learn the MWM task at 4 weeks post-injury. Hemispheric tissue loss and cortical lesion volume were similar among the brain-injured genotypes. Neither TBI nor the absence of NogoA/B caused an increased A beta expression. Myelin staining showed a reduced area and density in the corpus callosum in brain injured Nogo-A/B(-/-) animals compared to their littermate controls. These novel and unexpected behavioral results demonstrate that the absence of Nogo-A/B may negatively influence outcome, possibly related to hypomyelination, following TBI in mice and suggest a complex role for this myelin-associated axonal growth inhibitor following TBI. PMID- 19555746 TI - Whom do the sparrows follow? The effect of kinship on social preference in house sparrow flocks. AB - Non-aggressive social interactions between group-mates, e.g. maintenance of spatial proximity or activity synchrony are basic elements of a species' social structure, and were found to be associated with important fitness consequences in group-living animals. In the establishment of such affiliative relationships, kinship has often been identified as one of the key predictors, but this has rarely been studied in simple social groups such as flocks of gregarious birds. In this study we investigated whether kinship affects social preference, as measured by the tendency to associate with others during various social activities, in captive house sparrow (Passer domesticus) flocks where birds could interact with differently related flock-mates. We found that preference between flock-mates was correlated with familiarity from early nestling period: same brood siblings followed their sib initiating new activities more often than non sib birds. The strength of association between birds also tended to correlate with genetic relatedness, but this was mainly due to the effect of siblings' affiliation. Thus we concluded that house sparrows prefer the company of their siblings during social activities even well after fledging, which may facilitate kin-biased behaviours. PMID- 19555748 TI - Novel chronotherapeutic rectal aminophylline delivery system for therapy of asthma. AB - The aim of this study was to develop a new chronotherapeutic pharmaceutical preparation as a sustained-release suppository for prevention and therapeutic use against bronchial asthma in the early morning. Sustained-release hollow-type (SR HT) suppositories using sodium alginate (Alg-Na), sodium polyacrylate (PANa) or polyacrylate-PANa co-polymer (PA-PANa) as gelling polymers (gel agent) were prepared and pharmaceutical characteristics of these suppositories were investigated. Type A SR-HT suppositories comprised a suppository shell prepared with oleaginous base and containing aminophylline only or aminophylline with Alg Na or PANa in the cavity (hollow space). Type B SR-HT suppositories comprised a suppository shell prepared with oleaginous base and gel agent (30%), with aminophylline in the hollow space. In drug-release studies, the acrylate polymer containing suppositories showed linearity of delayed release rate, providing significantly decreased the highest concentration of theophylline in plasma (C(max)) and delayed the time required to reach C(max) (t(max)) and the mean residence time (MRT) after rectal administrated in rabbits. In particular, suppositories containing PA-PANa maintained significantly higher theophylline concentrations than control suppositories at 12h after rectal administration. Furthermore, histopathological examination indicated that these suppositories using acrylate polymers did not result in rectal lesions. The SR-HT suppository, particularly using PA-PANa as a gel agent, may thus be useful against nocturnal symptoms of asthma. In this study, we confirmed new formulation of sustained release suppository for chronotherapy of theophylline using oily base material in combination with polymer such as PA-PANa. The hollow-type suppository containing oleaginous base and hydrophilic polymer in the shell could be useful device for rectal administration of various drugs with prolongation of plasma concentration. PMID- 19555750 TI - Nanoparticles for direct nose-to-brain delivery of drugs. AB - This review aims to evaluate the evidence for the existence of a direct nose-to brain delivery route for nanoparticles administered to the nasal cavity and transported via the olfactory epithelium and/or via the trigeminal nerves directly to the CNS. This is relevant in the field of drug delivery as well as for new developments in nanotechnology. Experiments in animal models have shown that nano-sized drug delivery systems can enhance nose-to-brain delivery of drugs compared to equivalent drug solutions formulations. Protection of the drug from degradation and/or efflux back into the nasal cavity may partly be the reason for this effect of nanoparticles. It is uncertain, however, whether drug from the nanoparticles is being released in the nasal cavity or the nanoparticles carrying the drug are transported via the olfactory system or the trigeminal nerves into the CNS where the drug is released. Furthermore, toxicity of nanoparticulate drug delivery systems in the nasal cavity and/or in the CNS has not been extensively studied and needs to be considered carefully. PMID- 19555749 TI - Effect of sodium caprate on the oral absorptions of danshensu and salvianolic acid B. AB - The current study aims to investigate the effect of sodium caprate on the intestinal absorption and bioavailabilities of danshensu and salvianolic acid B, the major active components in Salvia miltiorrhiza Bge (Danshen). Biopharmaceutics and pharmacokinetics properties of the two compounds have been characterized by in vitro, in situ models as well as in vivo in rats. Based on the identified biopharmaceutics characteristics of the two compounds, effect of sodium carparate as absorption enhancer on the intestinal absorption and pharmacokinetics of danshensu and salvianolic acid B in pure compound form as well as extract form were investigated both in vitro and in vivo. Both danshensu and salvianolic acid B demonstrated very limited intestinal permeabilities, leading to oral bioavailabilities of only 11.09% and 3.90% in rats, respectively. Results from both in vitro and in vivo studies consistently indicated that sodium caprate could significantly enhance intestinal permeabilities as well as the in vivo bioavailabilities of both danshensu and salvianolic acid B. The current findings not only identified the usefulness of sodium caprate for the improved delivery of Danshen product but also demonstrated the importance of biopharmaceutics characterization in the dosage form development of traditional Chinese medicine. PMID- 19555747 TI - The adaptive role of transposable elements in the Drosophila genome. AB - Transposable elements (TEs) are short DNA sequences with the capacity to move between different sites in the genome. This ability provides them with the capacity to mutate the genome in many different ways, from subtle regulatory mutations to gross genomic rearrangements. The potential adaptive significance of TEs was recognized by those involved in their initial discovery although it was hotly debated afterwards. For more than two decades, TEs were considered to be intragenomic parasites leading to almost exclusively detrimental effects to the host genome. The sequencing of the Drosophila melanogaster genome provided an unprecedented opportunity to study TEs and led to the identification of the first TE-induced adaptations in this species. These studies were followed by a systematic genome-wide search for adaptive insertions that allowed for the first time to infer that TEs contribute substantially to adaptive evolution. This study also revealed that there are at least twice as many TE-induced adaptations that remain to be identified. To gain a better understanding of the adaptive role of TEs in the genome we clearly need to (i) identify as many adaptive TEs as possible in a range of Drosophila species as well as (ii) carry out in-depth investigations of the effects of adaptive TEs on as many phenotypes as possible. PMID- 19555751 TI - Enhancement of gene transfection into human dendritic cells using cationic PLGA nanospheres with a synthesized nuclear localization signal. AB - Effective delivery of DNA encoding antigen into the dendritic cells (DCs), which are non-dividing cells, is very important for the development of DNA vaccines. In a previous study, we developed the PLGA nanospheres that contained a cationic nanomaterial and showed high transfection efficiency in COS7 cells, which divide. In the present study, to produce an effective vector for the DNA vaccines, the gene expression and intracellular trafficking of pDNA complexed with PLGA/PEI nanospheres, in combination with an NF-kappaB analog as a nuclear localization signal (NLS) and electroporation were evaluated in human monocyte-derived DCs (hMoDCs). Cellular uptake of pDNA both in COS7 cells and hMoDCs was enhanced using the PLGA/PEI nanospheres. On the other hand, the PLGA/PEI nanospheres significantly promoted the transfection in COS7 cells, but had almost no effect on transfection in hMoDCs. The intranuclear transport of pDNA by PLGA/PEI nanospheres in COS7 cells was significantly higher than that in hMoDCs. These results indicate that pDNA complexed with PLGA/PEI nanospheres cannot enter into the nuclei of non-dividing cells. However, PLGA/PEI nanospheres combinated with NLS and electroporation (experimental permeation enhancer) greatly elevated the transfection efficiency by improvement of not only intracellular uptake but also intranuclear transport of pDNA in the hMoDCs. Thus, this delivery system using nanospheres combined with synthesized NLS might be applicable to DC-based gene vaccines when much non-invasive application such as needle-free injector should be required. PMID- 19555752 TI - The effects of common solubilizing agents on the intestinal membrane barrier functions and membrane toxicity in rats. AB - The use of solubilizing agents to improve the solubility of poorly water-soluble drugs often results in an alteration of intestinal membrane barrier function and intestinal membrane damage. In this study, 5(6)-carboxyfluorescein (CF) and fluorescein isothiocyanate-labeled dextran (MW 4400, FD4) were used as model compounds to examine the effects of twelve common solubilizing agents, sodium taurocholate (NaTC), Labrasol, polyethylene glycol 400 (PEG 400), Transcutol P, propylene glycol, Gelucire 44/14, HCO-60, ethanol, Cremophor EL, Tween 80, 2 hydroxypropyl-beta-cyclodextrin (2HP-beta-CyD) and dimethylsulfoxide (DMSO), on intestinal membrane barrier function and membrane toxicity in rats. Intestinal transport and absorption of CF were examined using an in vitro diffusion chamber and an in situ closed-loop technique. The in vitro diffusion chamber study showed that only 5 and 10% (w/v) NaTC significantly increased the transport of CF across the intestinal membrane. The in situ closed-loop study showed a remarkable increase in the absorption of CF and a bioavailability of more than 30% in the presence of 5 and 10% (v/v) Labrasol, 5 and 10% (w/v) NaTC and 10% (v/v) Transcutol P. Furthermore, we evaluated the effect of NaTC and Labrasol on the intestinal absorption of FD4, a high molecular weight compound. The results indicated that the absorption of FD4 also increased in the presence of 5 and 10% (w/v) NaTC and 10% (v/v) Labrasol, suggesting that these concentrations of NaTC and Labrasol may alter the intestinal membrane barrier functions in rats. We measured the release of protein and lactate dehydrogenase (LDH) from the intestinal membrane to examine the safety of solubilizing agents in the intestine. 5 and 10% (w/v) NaTC and 5 and 10% (v/v) Gelucire 44/14 significantly increased the presence of these toxicity markers compared to the control. The LDH level was also increased in the presence of 10% (v/v) of Cremophor EL. These findings suggest that the solubilizing agents at these concentrations except for NaTC, Gelucire 44/14 and Cremophor EL are considered safe and do not cause intestinal membrane damage. In conclusion, this study provides a basic approach in screening and predicting the effects of solubilizing agents for intestinal absorption studies using drugs poorly soluble in water. PMID- 19555753 TI - Electrophysiological and psychophysical asymmetries in sensitivity to interaural correlation steps. AB - The binaural auditory system's sensitivity to changes in the interaural cross correlation (IAC), as an indicator for the perceived spatial diffuseness of a sound, is of major importance for the ability to distinguish concurrent sound sources. In this article, we present electroencephalographical and corresponding psychophysical experiments with stepwise transitions of the IAC in continuously running noise. Both the transient and sustained brain response, display electrophysiological correlates of specific binaural processing in humans. The transient late auditory evoked potentials (LAEP) systematically depend on the size of the IAC transition, the reference correlation preceding the transition, the direction of the transition and on unspecific context information from the stimulus sequence. The psychophysical and electrophysiological data are characterized by two asymmetries. (1) Major asymmetry: for reference correlations of +1 and -1, psychoacoustical thresholds are comparatively lower, and the peak to-peak-amplitudes of LAEP are larger than for a reference correlation of zero. (2) Minor asymmetry: for IAC transitions in the positive parameter range, perceptual thresholds are slightly better and peak-to-peak amplitudes are larger than in the negative range. In all experimental conditions, LAEP amplitudes are linearly related to the dB scaled power ratio of correlated (N(0)) versus anticorrelated (N(pi)) signal components. The voltage gain of LAEP per dB(N(0)/N(pi)) closely corresponds to a constant perceptual distance between two correlations. We therefore suggest that activity in the auditory cortex and perceptual IAC sensitivity are better represented by the dB-scaled N(0)/N(pi) power ratio than by the normalized IAC itself. PMID- 19555754 TI - Detection of RNA expression from pseudogenes and non-coding genomic regions of Mycobacterium leprae. AB - We have previously reported that some pseudogenes are expressed in Mycobacterium leprae (M. leprae), the causative agent of leprosy, and that their expression levels alter upon infection of macrophages. We attempted to further examine the expression of pseudogene and non-coding genomic region in M. leprae, in this study. 19 Pseudogenes, 17 non-coding genomic regions, and 21 coding genes expression in M. leprae maintained in the footpads of the hypertensive nude rat (SHR/NCrj-rnu) were examined by reverse transcriptase polymerase chain reaction (RT-PCR). The expression of some of these pseudogenes, non-coding genomic regions and coding genes were also examined in M. leprae from skin smear specimens obtained from patients with lepromatous leprosy by RT-PCR. Transcripts from pseudogenes, non-coding genomic regions and coding genes examined in this study were clearly observed in M. leprae. The expression patterns of some of these transcripts vary greatly among different leprosy patients. These results indicate that some of pseudogenes and non-coding genomic regions are transcribed in M. leprae and analysis of RNA expression patterns including pseudogene and non coding genomic region in M. leprae may be useful in understanding the pathological states of infected patients. PMID- 19555755 TI - Sheep (Ovis aries) integrins alphavbeta1 and alphavbeta6 related to foot-and mouth disease virus infection: molecular cloning, sequence analysis and comparison with homologues. AB - Four members of the alphav integrin family of cellular receptors, alphavbeta1, alphavbeta3, alphavbeta6, and alphavbeta8, have been identified as receptors for foot-and-mouth disease virus (FMDV) in vitro, and integrins are believed to be the receptors used to target epithelial cells in the infected animals. To analyse roles of the alphav integrins from a susceptible species as viral receptors, we have cloned sheep alphav, beta1, and beta6 integrin cDNAs and compared them to those of other species. The coding sequences for sheep integrin alphav, beta1, and beta6 were found to be 3147, 2397, and 2364 nuclotides in length, encoding 1048, 798, and 787 amino acids, respectively. The sheep alphav, beta1, and beta6 subunits share many structural features including ligand binding domain and cysteine-rich region with homologues of other species. Phylogenetic trees and similarity analyses showed the close relationship of integrin genes among sheep, pigs, cattle and Bactrian camels that are susceptible to FMDV infection, which were distinct from the order Rodentia, Primates, Perissodactyla, Carnivora, Galliformes. We postulate that host tropism of FMDV may be related to divergence in integrin subunits among different species. PMID- 19555756 TI - The expression of the Goodpasture antigen-binding protein (ceramide transporter) in adult rat brain. AB - The Goodpasture antigen-binding protein (GPBP) plays a critical role in brain development. Knockdown of GPBP leads to loss of myelinated tracts in the central nervous system and to extensive apoptosis in the brain during early embryogenesis. GPBP was initially identified as a protein associated with the autoantigen in Goodpasture autoimmune syndrome, where it was shown to be a kinase that regulates type IV collagen organization. GPBP isoforms bind and transport ceramide from the endoplasmic reticulum to the Golgi apparatus and are therefore also known as ceramide transporters (CERT). Ceramide dysregulation is involved in autoimmunity and neurodegenerative disorders. In order to analyze the possible role of GPBP in neuroinflammation and neurodegeneration we studied the basal GPBP expression in normal rat brain. High levels of immunoreactivity were detected in neurons of the cerebral cortex, hippocampal formation, the basal ganglia, the olfactory bulb and nuclei of the thalamus, the hypothalamus and the septal area. Lower expression levels of GPBP were observed widely throughout the brain, suggesting that GPBP plays an important role in central nervous system neuron function. PMID- 19555757 TI - Correlation of F4-neuroprostanes levels in cerebrospinal fluid with outcome of aneurysmal subarachnoid hemorrhage in humans. AB - Aneurysmal subarachnoid hemorrhage (aSAH) is one type of hemorrhagic stroke in humans. F(2)-isoprostanes (F(2)-IsoPs) and F(4)-neuroprostanes (F(4)-NPs), derived from arachidonic acid and docosahexaenoic acid (DHA), respectively, are specific markers of lipid peroxidation. We previously demonstrated that F(2) IsoPs levels in cerebrospinal fluid (CSF) of aSAH patients positively correlated with poor clinical conditions. In this work, we refined F(4)-NPs analysis and investigated the role of potential oxidative damage to neurons in aSAH patients by detecting F(4)-NPs in CSF. [(2)H(4)]-15-F(2t)-IsoP, rather than [(18)O(2)]-17 F(4c)-NP or [(2)H(4)]-PGF(2 alpha), was used as the internal standard for F(4) NPs analysis. One problem of the use of [(18)O(2)]-17-F(4c)-NP was the potential interference resulting from F(2)-dihomo-IsoPs in CSF. CSF specimens of 15 aSAH patients for up to 10 days and those of 12 non-aSAH controls were analyzed. First day, mean, and peak levels of F(4)-NPs were all significantly higher in aSAH patients than in controls and correlated with the Fisher Scale and 3-month Glasgow Outcome Scale, but only mean levels of F(4)-NPs correlated with Hunt and Hess Grade. The results first demonstrate oxidative damage to DHA in brain tissue following aSAH and suggest that F(4)-NPs in CSF could be a better predictor for outcome of aSAH than F(2)-IsoPs at early time points. PMID- 19555759 TI - Activation of caspase-8 by tumour necrosis factor receptor 1 is necessary for caspase-3 activation and apoptosis in oxygen-glucose deprived cultured cortical cells. AB - TNF-alpha has been reported to be relevant in stroke-induced neuronal death. However the precise function of TNF-alpha in brain ischemia remains controversial since there are data supporting either a detrimental or a protective effect. Here we show that TNF-alpha is released after oxygen-glucose deprivation (OGD) of cortical cultures and is a major contributor to the apoptotic death observed without affecting the OGD-mediated necrotic cell death. In this paradigm, apoptosis depends on TNF-alpha-induced activation of caspase-8 and -3 without affecting the activation of caspase-9. By using knock-out mice for TNF-alpha receptor 1, we show that the activation of both caspase-3 and -8 by TNF-alpha is mediated by TNF-alpha receptor 1. The pro-apoptotic role of TNF-alpha in OGD is restricted to neurons and microglia, since astrocytes do not express either TNF alpha or TNF-alpha receptor 1. Altogether, these results show that apoptosis of cortical neurons after OGD is mediated by TNF-alpha/TNF-alpha receptor 1. PMID- 19555760 TI - BDNF regulates BIM expression levels in 3-nitropropionic acid-treated cortical neurons. AB - 3-Nitropropionic acid (3-NP) is an irreversible inhibitor of succinate dehydrogenase that has been used to explore the primary mechanisms of cell death associated with mitochondrial dysfunction and neurodegeneration in Huntington's disease. In this study we investigated the ability of brain-derived neurotrophic factor (BDNF) to suppress mitochondrial-dependent cell death induced by 3-NP in primary cortical neurons. This neurotrophin prevented 3-NP-induced release of cytochrome c and Smac/Diablo, caspase-3-like activity and nuclear condensation/fragmentation. Furthermore, it greatly increased phosphorylation of Akt and MAPK, suggesting the involvement of these signalling pathways in BDNF neuroprotection. Interestingly, BDNF decreased the levels of the pro-apoptotic protein Bim in mitochondrial and total cell lysates through the activation of the MEK1/2 pathway. This effect was due to an increase in the degradation rates of Bim. Our data support an important role for BDNF, in protecting cortical neurons against apoptotic cell death caused by inhibition of mitochondrial complex II. PMID- 19555761 TI - P90 Ribosomal s6 kinase 2 negatively regulates axon growth in motoneurons. AB - Mutations in Ribosomal s6 kinase 2 (Rsk2) are associated with severe neuronal dysfunction in Coffin-Lowry syndrome (CLS) patients, flies and mice. So far, the mechanisms of how Rsk2 regulates development, maintenance and activity of neurons are not understood. We have investigated the consequences of Rsk2 deficiency in mouse spinal motoneurons. Survival of isolated Rsk2 deficient motoneurons is not reduced, but these cells grow significantly longer neurites. Conversely, overexpression of a constitutively active form of Rsk2 leads to reduced axon growth. Increased axon growth in Rsk2 deficient neurons was accompanied by higher Erk 1/2 phosphorylation, and the knockout phenotype could be rescued by pharmacological inhibition of MAPK/Erk kinase (Mek). These data indicate that Rsk2 negatively regulates axon elongation via the MAPK pathway. Thus, the functional defects observed in the nervous system of CLS patients and animal models with Rsk2 deficiency might be caused by dysregulated neurite growth rather than primary neurodegeneration. PMID- 19555758 TI - Image analysis of Ca2+ signals as a basis for neurotoxicity assays: promises and challenges. AB - Free intracellular calcium ([Ca(2+)](i)) controls a wide range of cellular functions such as contraction, neurotransmitter and hormone release, metabolism, cell division and differentiation. Cytosolic Ca(2+) levels are abnormal in cells exposed to toxicants and understanding how these levels become altered may improve our ability to design high-throughput methods for the sensitive detection of cellular responses to a toxic exposure. Because Ca(2+) is involved in multiple aspects of cellular function, its role in signaling is complex. It is therefore necessary to identify the individual pathways targeted during toxicant exposure in order to use them as a tool for predictive measurements of toxicity and as targets for prevention or reversal of injury. This review illustrates several methods available for analysis of Ca(2+) responses in vitro and their applicability for understanding mechanisms of toxicity at the molecular and cellular levels. The review will also consider the usefulness of Ca(2+) imaging for predicting a unique signature for classes of toxicants. Towards this end, two methodological approaches for assessment of Ca(2+) responses to toxicants are examined: steady state measurements and complex spatial and/or temporal measurements. Each of the methods described and appropriately used results in reliable and reproducible measurements which may be applied in a high-throughput fashion to individualize in vitro assessment of cellular responses caused by toxicants. PMID- 19555762 TI - Sp4-dependent repression of neurotrophin-3 limits dendritic branching. AB - Regulation of neuronal gene expression is critical to establish functional connections in the mammalian nervous system. The transcription factor Sp4 regulates dendritic patterning during cerebellar granule neuron development by limiting branching and promoting activity-dependent pruning. Here, we investigate neurotrophin-3 (NT3) as a target gene important for Sp4-dependent dendritic morphogenesis. We found that Sp4 overexpression reduced NT3 promoter activity whereas knockdown of Sp4 increased NT3 promoter activity and mRNA. Moreover, Sp4 bound to the NT3 promoter in vivo, supporting a direct role for Sp4 as a repressor of NT3 expression. Addition of exogenous NT3 promoted dendritic branching in cerebellar granule neurons. Furthermore, sequestering NT3 blocked the continued addition of dendritic branches observed upon Sp4 knockdown, but had no effect on dendrite pruning. These findings demonstrate that, during cerebellar granule neuron development, Sp4-dependent repression of neurotrophin-3 is required to limit dendritic branching and thereby promote acquisition of the mature dendritic pattern. PMID- 19555763 TI - Expression, purification and characterization of recombinant Z alpha(1) antitrypsin--the most common cause of alpha(1)-antitrypsin deficiency. AB - Alpha(1)-antitrypsin (alpha(1)AT), the most abundant proteinase inhibitor circulating in the blood, protects extracellular matrix proteins of the lung against proteolytic destruction by neutrophil elastase. alpha(1)AT deficiency predisposes patients to emphysema, juvenile cirrhosis and hepatocellular carcinoma. Over 90% of clinical cases of severe alpha(1)AT deficiency are caused by the Z variant (E342K) of alpha(1)AT. The presence of the Z mutation results in misfolding and polymerization of alpha(1)AT. Due to its inherent propensity to polymerize there are no reported cases of recombinant Z alpha(1)AT production. This has created a major impediment to studying the effect of the Z mutation on alpha(1)AT. Here we report our attempts to produce recombinant Z alpha(1)AT using both Escherichia coli and Pichia pastoris as host systems. Using a range of expression vectors in E. coli we were unable to produce soluble active Z alpha(1)AT. Cytosolic expression of the Z alpha(1)AT gene in P. pastoris was successful. Monomeric and active recombinant Z alpha(1)AT was purified from the yeast cytosol using affinity chromatography and anion exchange chromatography. Biochemical analyses demonstrated that the recombinant Z alpha(1)AT has identical properties to its native counterpart purified from plasma of patients homozygous for the Z allele. A recombinant source of pathological Z alpha(1)AT will increase the chances of elucidating the mechanism of its polymerization and thus the development of therapeutic strategies. PMID- 19555764 TI - Automatic particle selection from electron micrographs using machine learning techniques. AB - The 3D reconstruction of biological specimens using Electron Microscopy is currently capable of achieving subnanometer resolution. Unfortunately, this goal requires gathering tens of thousands of projection images that are frequently selected manually from micrographs. In this paper we introduce a new automatic particle selection that learns from the user which particles are of interest. The training phase is semi-supervised so that the user can correct the algorithm during picking and specifically identify incorrectly picked particles. By treating such errors specially, the algorithm attempts to minimize the number of false positives. We show that our algorithm is able to produce datasets with fewer wrongly selected particles than previously reported methods. Another advantage is that we avoid the need for an initial reference volume from which to generate picking projections by instead learning which particles to pick from the user. This package has been made publicly available in the open-source package Xmipp. PMID- 19555765 TI - Influence of probiotic feeding duration on disease resistance and immune parameters in rainbow trout. AB - The effect of feeding the probiotic Kocuria SM1 to rainbow trout (Oncorhynchus mykiss, Walbaum) on disease resistance was evaluated. Thus, rainbow trout were fed Kocuria SM1 supplemented diets at concentrations of approximately 10(8) cells g(-1) feed for up to four weeks, and then challenged intraperitoneally with Vibrio anguillarum at weekly intervals. A two-week feeding regime led to the maximum reduction in mortalities, i.e. 16%, compared to mortalities of 62, 30 and 22% for one, three and four week feeding regimes, respectively. These compared to 70-90% mortalities of the controls. An enhanced cellular and humoral immune response, notably greater head kidney macrophage phagocytic and peroxidase activities, and higher serum lysozyme and total protein levels were recorded after two weeks of probiotic administration. These results reveal that a two-week feeding regime with Kocuria SM1 leads to higher disease protection in rainbow trout, with protection linked to stimulation of immune parameters. PMID- 19555766 TI - Intraspecific differentiation of Limonium wrightii (Plumbaginaceae) on northwestern Pacific Islands: rate heterogeneity in nuclear rDNA and its distance independent geographic structure. AB - We conducted a molecular phylogenetic analysis of a coastal sea lavender in NW Pacific using ITS sequences of nrDNA. As a result, the ITS sequences of populations on the oceanic Daito Islands were considerably different from conspecific others in nine apomorphic substitutions. This finding indicates a distance-independent geographical structure of the genetic variation among islands in NW Pacific. The calibrated rate of nucleotide substitutions in the ITS regions within L. wrightii (average: 1.41x10(-8) substitutions per site per year) was higher than any other instances of substitution rates in flowering plants. PMID- 19555767 TI - Prognostic implication of CDC25A and cyclin E expression on primary breast cancer patients. AB - Defect in cell cycle control is a hallmark character of cancer. We have investigated the association of Ki67 labeling index, cyclin E and CDC25A expressions with clinical follow-up data in breast carcinomas. Flow cytometry was used to detect gene amplification of cyclins in 44 tumor tissue with invasive breast carcinomas. Multivariate Cox proportional hazard ratio test was used to show the correlations. Cyclin E or CDC25A were upregulated in 34% of the tumors. Among the whole total material, expression of cyclin E and of CDC25A were found upregulated in 31.9% and 39.4% of cells, respectively. Both CDC25A and cyclin E protein expression levels were correlated with Ki67 expression level (p<0.001). In addition, the expression of CDC25A was associated significantly with poor survival (P=0.028), whereas no correlation was found with cyclin E. These findings suggest a possible prognostic value for CDC25A as a cell cycle marker and may imply in characteristic of high risk breast cancer patients. PMID- 19555768 TI - Juglone, a naphthoquinone from walnut, exerts cytotoxic and genotoxic effects against cultured melanoma tumor cells. AB - This study demonstrates cytotoxic and genotoxic potential of juglone, a chief constituent of walnut, and its underlying mechanisms against melanoma cells. MTT assay and clonogenic assay were used to study cytotoxicity, micronucleus assay to assess genotoxicity, glutathione (GSH) assay and 2',7'-dicholorofluorescein diacetate (DCFH-DA) assay to evaluate the oxidative stress induction. Apoptosis/necrosis induction was analysed by flow cytometry. We observed a concentration-dependent decrease in cell survival with a corresponding increase in the lactate dehydrogenase levels. A dose-dependent increase in the frequency of micronucleated binucleate cells indicated the potential of juglone to induce cytogenetic damage in melanoma tumor cells. Moreover, results of the micronuclei study indicated division delay in the proliferating cell population by showing decrease in the cytokinesis blocked proliferation index. Further, juglone-induced apoptosis and necrosis could be demonstrated by oligonucleosomal ladder formation, microscopic analysis, increase in the hypodiploid fraction (sub Go peak in DNA histogram), as well as an increased percentage of AnnexinV(+)/PI(+) cells detected by flow cytometry. A significant concentration-dependent decrease in the glutathione levels and increase in dichlorofluorescein (DCF) fluorescence after juglone treatment confirmed the ability of juglone to generate intracellular reactive oxygen species. The cytotoxic effect of juglone can be attributed to mechanisms including the induction of oxidative stress, cell membrane damage, and a clastogenic action leading to cell death by both apoptosis and necrosis. PMID- 19555769 TI - Scratch wound closure of C2C12 mouse myoblasts is enhanced by human platelet lysate. AB - The effect of a platelet lysate (PL) on muscle wound healing, based on in vitro scratch wound of C2C12 mouse myoblasts, has been investigated. Cell viability assays show that PL induced an increase in cell proliferation at concentrations of 1-20%, but was slightly cytotoxic at 100%. PL promoted wound closure after scratch wounding of cell monolayers. The p38 inhibitor SB203580 and the PI3K inhibitor, wortmannin, decreased the PL effect, whereas the ERK inhibitor, PD98059, did not. Transwell migration of cells was also increased by PL, and although SB203580 abrogated this effect, wortmannin reduced it, whereas PD98059 was ineffective. Western blot analyses of scratch wounded cells showed activation of AKT and p38, while in the presence of PL there was a faster and sustained activation of AKT and p38 (up to 6h), and a transient activation of ERK1/2. Taken together, the data show that PL promotes C2C12 wound healing by enhancing cell proliferation and motility. PMID- 19555770 TI - Expression analysis of Fgf8a &Fgf8b in early stage of P19 cells during neural differentiation. AB - Fgf8 is a member of the fibroblast growth factor (FGF) family that plays an important role in early neural development. Cellular aggregation and retinoic acid (RA) are needed for mouse embryonic carcinoma (EC) P19 cell neural differentiation. We have examined the Fgf8 gene in P19 cells during neural differentiation and identified 2 alternatively spliced Fgf8 isoforms, Fgf8a and Fgf8b, among the 8 known splicing isoforms in mammals. The expression of Fgf8a and Fgf8b mRNAs transiently and rapidly increased in the early stage of P19 cells during RA-induced neural differentiation, followed by a decline in expression. The relative amount of Fgf8b was clearly higher than that of Fgf8a at different time-points measured within 24h after RA treatment. Increased Fgf8b mRNA expression was cellular-aggregation dependent. The results demonstrated that cellular-aggregation-induced Fgf8b, but not Fgf8a, may play a pivotal role in early neural differentiation of P19 cells. PMID- 19555772 TI - Ultraviolet radiation and Vitamin D3 in amphibian health, behaviour, diet and conservation. AB - Amphibians are currently suffering a period of mass extinction with approximately 20% of species under severe threat and more than 120 species already extinct. In light of this crisis there is an urgency to establish viable ex situ populations and also find the causes of in situ declines. The role of ultraviolet radiation and Vitamin D(3) in amphibian health directly influences both ex situ and in situ populations. Vitamin D(3) can be photosynthesised endogenously via UV-B radiation (UV-B), or acquired through the diet, and then metabolised to calcitriol the biologically active hormonal form. Although, there is a lack of literature concerning Vitamin D(3) requirements and calcitriol synthesis in amphibians, amphibians are likely to have similar Vitamin D(3) requirements and metabolic processes as other vertebrates due to the phylogenetically conservative nature of calcitriol biosynthesis. Deficiencies in calcitriol in amphibians result in nutritional metabolic bone disease (NMBD) and could compromise reproduction and immunity. However, excess biologically active UV radiation has also proven detrimental across all three amphibian life stages and therefore could impact both in situ and ex situ populations. Here we review the role and necessity of UV B and calcitriol in amphibians and the potential for negative impacts due to excessive exposure to UV radiation. We also identify priorities for research that could provide critical information for maintaining healthy in ex situ and in situ populations of amphibians. PMID- 19555773 TI - Tissue-specific and embryonic expression of the retinoid X receptors in Sebastiscus marmoratus. AB - Retinoid X receptors (RXRs) are highly conserved members of the nuclear receptor family and mediate various physiological processes in vertebrates. Most studies on RXRs have concentrated on their structure and function in mammals and their characterization and developmental expression in Danio rerio. However, there is little information concerning the distribution of RXRs in teleost tissues. In the present study, we cloned partial sequences of three RXR subtypes (RXRa, -b, -g) from Sebastiscus marmoratus by RACE PCR and analyzed the phylogeny of the teleost and the tetrapod RXR genes, and identified some inconsistencies with previous studies. The tissue-specific and embryonic expression profiles of each RXR gene were explored using real time quantitative PCR. This analysis demonstrated that these RXRs were expressed in all test tissues indicating their participation in many physiological processes. However, we found a great difference in the distribution of RXRg between teleosts and mammals. Furthermore, we followed expression of the three subtypes through various embryo developmental stages and found that the RXRa orthologues of teleosts might be involved in the development of the anterior hindbrain, tailbud and neural crest and in the formation of the pharynx and fin, that RXRb played ubiquitous roles in fish early development, and that RXRg probably played a role in brain and nervous system development and function. PMID- 19555771 TI - Trifluoromethanesulfonic acid-based proteomic analysis of cell wall and secreted proteins of the ascomycetous fungi Neurospora crassa and Candida albicans. AB - Cell wall proteins from purified Candida albicans and Neurospora crassa cell walls were released using trifluoromethanesulfonic acid (TFMS) which cleaves the cell wall glucan/chitin matrix and deglycosylates the proteins. The cell wall proteins were then characterized by SDS-PAGE and identified by proteomic analysis. The analyses for C. albicans identified 15 cell wall proteins and six secreted proteins. For N. crassa, the analyses identified 26 cell wall proteins and nine secreted proteins. Most of the C. albicans cell wall proteins are found in the cell walls of both yeast and hyphae cells, but some cell type-specific cell wall proteins were observed. The analyses showed that the pattern of cell wall proteins present in N. crassa vegetative hyphae and conidia (asexual spores) are quite different. Almost all of the cell wall proteins identified in N. crassa have close homologs in the sequenced fungal genomes, suggesting that these proteins have important conserved functions within the cell wall. PMID- 19555774 TI - Quantification of statin effects on hepatic cholesterol synthesis by transient (13)C-flux analysis. AB - The present work is the first to deal with the determination of cholesterol synthesis rates in primary rat hepatocytes using transient (13)C-flux analysis. The effects of statins on cholesterol biosynthesis and central carbon fluxes were quantified at a therapeutic concentration of 50 nM atorvastatin using carbon labeled glutamine. The flux through the cholesterol pathway decreased from 0.27 to 0.08 mmol/l(cv)h in response to the administration of the hypolipidemic drug. Isotopic steady state was reached within 4h in the central carbon metabolism but not in the cholesterol pathway, regardless of whether atorvastatin was administered or not. Marked channeling was observed for the symmetrical tricarboxylic acid cycle intermediates, succinate and fumarate. Non-stationary (13)C-based flux identification delivers both intracellular fluxes and intermediate levels, which was for the first time utilized for investigating systems-level effects of the administered drug by quantifying the flux control of the 3-hydroxy-3-methylglutaryl-coenzyme A reductase. PMID- 19555778 TI - Identification and biochemical characterization of a GDSL-motif carboxylester hydrolase from Carica papaya latex. AB - An esterase (CpEst) showing high specific activities on tributyrin and short chain vinyl esters was obtained from Carica papaya latex after an extraction step with zwitterionic detergent and sonication, followed by gel filtration chromatography. Although the protein could not be purified to complete homogeneity due to its presence in high molecular mass aggregates, a major protein band with an apparent molecular mass of 41 kDa was obtained by SDS-PAGE. This material was digested with trypsin and the amino acid sequences of the tryptic peptides were determined by LC/ESI/MS/MS. These sequences were used to identify a partial cDNA (679 bp) from expressed sequence tags (ESTs) of C. papaya. Based upon EST sequences, a full-length gene was identified in the genome of C. papaya, with an open reading frame of 1029 bp encoding a protein of 343 amino acid residues, with a theoretical molecular mass of 38 kDa. From sequence analysis, CpEst was identified as a GDSL-motif carboxylester hydrolase belonging to the SGNH protein family and four potential N-glycosylation sites were identified. The putative catalytic triad was localised (Ser(35)-Asp(307) His(310)) with the nucleophile serine being part of the GDSL-motif. A 3D-model of CpEst was built from known X-ray structures and sequence alignments and the catalytic triad was found to be exposed at the surface of the molecule, thus confirming the results of CpEst inhibition by tetrahydrolipstatin suggesting a direct accessibility of the inhibitor to the active site. PMID- 19555779 TI - Refolding, characterization and crystal structure of (S)-malate dehydrogenase from the hyperthermophilic archaeon Aeropyrum pernix. AB - Tartrate oxidation activity was found in the crude extract of an aerobic hyperthermophilic archaeon Aeropyrum pernix, and the enzyme was identified as (S) malate dehydrogenase (MDH), which, when produced in Escherichia coli, was mainly obtained as an inactive inclusion body. The inclusion body was dissolved in 6 M guanidine-HCl and gradually refolded to the active enzyme through dilution of the denaturant. The purified recombinant enzyme consisted of four identical subunits with a molecular mass of about 110 kDa. NADP was preferred as a coenzyme over NAD for (S)-malate oxidation and, unlike MDHs from other sources, this enzyme readily catalyzed the oxidation of (2S,3S)-tartrate and (2S,3R)-tartrate. The tartrate oxidation activity was also observed in MDHs from the hyperthermophilic archaea Methanocaldococcus jannaschii and Archaeoglobus fulgidus, suggesting these hyperthermophilic MDHs loosely bind their substrates. The refolded A. pernix MDH was also crystallized, and the structure was determined at a resolution of 2.9 A. Its overall structure was similar to those of the M. jannaschii, Chloroflexus aurantiacus, Chlorobium vibrioforme and Cryptosporidium parvum [lactate dehydrogenase-like] MDHs with root-mean-square-deviation values between 1.4 and 2.1 A. Consistent with earlier reports, Ala at position 53 was responsible for coenzyme specificity, and the next residue, Arg, was important for NADP binding. Structural comparison revealed that the hyperthermostability of the A. pernix MDH is likely attributable to its smaller cavity volume and larger numbers of ion pairs and ion-pair networks, but the molecular strategy for thermostability may be specific for each enzyme. PMID- 19555780 TI - Profiling of calpain activity with a series of FRET-based substrates. AB - Calpains are intracellular proteases that selectively cleave proteins in response to calcium signals. Although calpains cut many different sequences, residue preferences within peptide substrates were recently determined and incorporated into a superior FRET (fluorescence resonance energy transfer)-based substrate (PLFAER). Here we show PLFAER is cleaved by calpain at the intended F-A scissile bond. Sequential replacement of individual residues by alanine reduced activity except with PLFAAR, which is cleaved 2.3 times faster than PLFAER. The rates of hydrolysis of the alanine-substituted substrates were used to compare substrate preferences of calpain, papain and cathepsins B and L. The preferences of the two major isoforms, calpains 1 and 2, were virtually indistinguishable and were very similar to those of the calpain 1 protease core and papain. However, the activity profiles with the FRET substrate series were significantly different for the cathepsins, particularly cathepsin B. PMID- 19555781 TI - Molecular characterization of young and mature odontoblasts. AB - The odontoblast is the secretory cell responsible for primary, secondary and tertiary reactionary dentinogenesis. We provide evidence that the changes in secretory activity of odontoblasts reflect differential transcriptional control and that common regulatory processes may exist between dentine and bone. INTRODUCTION: Based on the hypothesis that differential dentine secretion (primary and secondary dentinogenesis) is associated with changes in the transcriptional control within the cell, we have investigated the transcriptome of odontoblasts at young and mature stages and subsequently used this information to identify key regulatory intracellular pathways involved in this process. MATERIALS AND METHODS: We used microarray analysis to compare the transcriptome of early stage (primary dentinogenesis) and late stage (secondary dentinogenesis) odontoblasts from 30 month old bovine teeth. Secondarily, we used post-array sqRT PCR to confirm the differential expression of 23 genes in both populations of odontoblasts. Finally, immunohistochemistry was performed on bovine and murine tissues with antibodies to DMP1 and anti-phospho p38 proteins. RESULTS: DMP-1 and osteocalcin gene expression were up-regulated in the mature odontoblasts, whereas collagen I, DSPP, TGF-beta1 and TGF-beta1R gene expression were down-regulated. Microarray analysis highlighted 574 differentially regulated genes (fold change>2 - p<0.05). This study supports further existing similarities between pulp cells and bone cells. Using post-array Sq-RT-PCR we characterized transcript levels of genes involved in the p38 MAP kinase pathway (PTPRR, NTRKK2, MAPK13, MAP2K6, MKK3). Differential p38 gene activation was confirmed by immunohistochemistry for p38 protein in murine teeth. Finally, immunohistochemistry for DMP1 indicated that odontoblasts involved in primary and secondary dentinogenesis may coexist in the same tooth. CONCLUSION: As established in bone cells, the transcriptome of the odontoblast was shown here to evolve with their stage and functional maturity. Identification of the involved signalling pathways, as highlighted for p38, will enable the deciphering of physiology and pathology of mineralised tissue formation. PMID- 19555783 TI - Reduced bone perfusion in proximal femur of subjects with decreased bone mineral density preferentially affects the femoral neck. AB - Using dynamic contrast enhanced MR imaging, this study investigated perfusion of the proximal femur in subjects with normal BMD, low bone mass and osteoporosis. Study cohort comprised healthy elderly Hong Kong Chinese volunteers consisting of 107 males (74.4+/-4.2 years, mean+/-SD) and 135 females (73.9+/-4.3 years). Right proximal femur BMD measurement by DXA and MR perfusion imaging (maximum enhancement, E(max) and enhancement slope, E(slope)) of the femoral head, neck, and proximal shaft were carried out within a one month interval. Normal BMD, low bone mass and osteoporotic subjects accounted for 46.7%, 44.9%, and 8.4% of males; and 32.6%, 43.7%, and 23.7% of females. Perfusion indices showed that femoral head perfusion was less perfused compared to the femoral shaft (E(max) and E(slope) indices of head region=28% of shaft region). Compared with normal BMD subjects, E(max) of femoral head, neck, and proximal femur shaft were reduced by 15+/-5% (mean+/-standard error); 40+/-4%; 15+/-5% respectively for low bone mass subjects, and 36+/-4%; 50+/-6%; 47+/-6% respectively for osteoporotic subjects. E(slope) of femoral head, neck, and proximal femur shaft were reduced by 17+/-7%; 41+/-5%; 4+/-7% for low bone mass subjects and 50+/-5%, 62+/-5%, 34+/ 8% for osteoporotic subjects. In low bone mass and osteoporotic subjects there was a tendency for perfusion in the femoral neck to reduce to a greater degree relative to that in the femoral head and shaft. PMID- 19555782 TI - Hypophosphatemia induced by intravenous administration of saccharated ferric oxide: another form of FGF23-related hypophosphatemia. AB - Fibroblast growth factor 23 (FGF23) is a humoral factor that is produced by osteocytes and regulates phosphate and vitamin D metabolism. Several hypophosphatemic diseases including X-linked, autosomal dominant and autosomal recessive hypophosphatemic rickets/osteomalacia and tumor-induced rickets/osteomalacia are caused by excess actions of FGF23. These diseases are characterized by hypophosphatemia associated with impaired proximal tubular phosphate reabsorption and inappropriately low serum 1,25-dihydroxyvitamin D [1,25(OH)(2)D] levels for hypophosphatemia. Saccharated ferric oxide is widely used in Japan for iron-deficiency anemia. While it has been shown that saccharated ferric oxide induces hypophosphatemic osteomalacia, the mechanism of this hypophosphatemia remains to be clarified. We here describe three hypophosphatemic patients caused by intravenous administration of saccharated ferric oxide. Hypophosphatemia in these patients were associated with impaired renal tubular phosphate reabsorption, rather low serum 1,25(OH)(2)D and high FGF23 levels. All these biochemical features improved by the cessation of saccharated ferric oxide. These results indicate that hypophosphatemia caused by saccharated ferric oxide is another form of FGF23-related hypophosphatemia. PMID- 19555784 TI - A simple single-port laparoscopic-assisted technique for completely enclosing inguinal hernia in children. AB - Single-port endoscopic-assisted percutaneous extraperitoneal closure of inguinal hernia with variable devices is a novel technique in minimal-access surgery for pediatric inguinal hernias. However, a small peritoneal gap of the suture at the location of spermatic vessels and vas deferens may be left untouched, whereas some subcutaneous tissues, including nerves and muscles, may cause injury by their inclusion in the upper portion of the circuit suturing. The technique may fail to entirely enclose the hernia defect and has the potential to lead to higher incidence of hydrocele and hernia recurrence. The authors describe the modification of single-port endoscopic-assisted ligation with a homemade hooked injection needle, which is designed to cause hydrodissection to the preperitoneal space, tautly enclose the hernia defect without upper subcutaneous tissues and a lower peritoneal gap, and aid in the achievement of a near-zero recurrence rate. PMID- 19555785 TI - Thoracoabdominal shotgun wounds: an evaluation of factors associated with the need for surgical intervention. AB - BACKGROUND: Shotgun wound classification systems attempt to predict the need for surgical intervention based on the size of wounds, pellet spread, or distance from the weapon rather than clinical findings. METHODS: A 5-year retrospective review of patients sustaining a thoracoabdominal shotgun wound was performed. Factors believed to be associated with the need for surgical intervention were examined using the Fisher exact test or an independent sample t test. RESULTS: Sixty-four patients suffered a thoracoabdominal shotgun wound. Fifty-nine percent required surgical intervention. Factors significantly associated with the need for surgical intervention were a low revised trauma score and systolic and diastolic blood pressure (P < .05). Distance from attacker, wound patterns, pellet size, and pellet spread were not found to have an association. CONCLUSIONS: Clinical indicators of hemorrhage and shock are associated with the need for surgical intervention, whereas pellet spread, pellet size, and distance from the attacker are not. This is a significant departure from traditional classification systems. PMID- 19555786 TI - Chronic abdominal wall pain--a diagnostic challenge for the surgeon. AB - BACKGROUND: Chronic abdominal wall pain (CAWP) occurs in about 30% of all patients presenting with chronic abdominal pain. METHODS: The authors review the literature identified in a PubMed search regarding the abdominal wall as the origin of chronic abdominal pain. RESULTS: CAWP is frequently misinterpreted as visceral or functional abdominal pain. Misdiagnosis often leads to a variety of investigational procedures and even abdominal operations with negative results. With a simple clinical test (Carnett's test), >90% of patients with CAWP can be recognized, without risk for missing intra-abdominal pathology. CONCLUSION: The condition can be confirmed when the injection of local anesthetics in the trigger point(s) relieves the pain. A fasciotomy in the anterior abdominal rectus muscle sheath through the nerve foramina of the affected branch of one of the anterior intercostal nerves heals the pain. PMID- 19555787 TI - Stress and addiction: a dynamic interplay of genes, environment, and drug intake. PMID- 19555788 TI - The "inactivity trap". PMID- 19555789 TI - The longitudinal effects of depression on physical activity. AB - OBJECTIVES: It has been posited that depression and sedentary lifestyle have bidirectional relationships. Although the role of baseline physical activity as a risk factor for emerging depression has been recently reviewed, there has been no systematic review of the literature to assess the reverse relationship. We reviewed the results of longitudinal studies in the world's literature that have studied the effect of baseline depression on ensuing levels of physical activity. MEASURES: MEDLINE, PsycINFO, CINHAL Plus, Health Source: Nursing Academic Edition and Cochrane databases were searched from 1959 to 2008 with a focus on depression, sedentary lifestyle and exercise. Published longitudinal studies in English and more than 100 patients were included. RESULTS: Eleven studies met our inclusion criteria, eight of which reported that baseline depression was significantly associated with subsequent sedentary lifestyle or poor adherence to the physical exercise regimens recommended by physicians after a coronary event. However, the studies used different depression scales and physical activity outcome measures, and varied a great deal in the range of potential confounders they controlled for. In addition, there were only three studies that were specifically designed to assess the role of baseline depression on the subsequent level of physical activity. CONCLUSIONS: Baseline depression may be a significant risk factor for development of sedentary lifestyle or decreased level of physical exercise. Future studies should examine mechanisms by which depression may lead to decline in activity. PMID- 19555790 TI - Was SARS a mental health catastrophe? PMID- 19555791 TI - Long-term psychiatric morbidities among SARS survivors. AB - OBJECTIVE: Severe acute respiratory syndrome (SARS) was the first massive infectious disease outbreak of the 21st century. However, it is unlikely that this outbreak will be the last. This study aimed to evaluate the long-term psychiatric morbidities in survivors of SARS. METHOD: This is a cohort study designed to investigate psychiatric complications among SARS survivors treated in the United Christian Hospital 30 months after the SARS outbreak. Psychiatric morbidities were assessed by the Structured Clinical Interview for DSM-IV, the Impact of Events Scale-Revised and the Hospital Anxiety and Depression Scale. Functional outcomes were assessed by the Medical Outcomes Study 36-Item Short Form Health Survey. RESULTS: Ninety subjects were recruited, yielding a response rate of 96.8%. Post-SARS cumulative incidence of DSM-IV psychiatric disorders was 58.9%. Current prevalence for any psychiatric disorder at 30 months post-SARS was 33.3%. One-fourth of the patients had post-traumatic stress disorder (PTSD), and 15.6% had depressive disorders. CONCLUSION: The outbreak of SARS can be regarded as a mental health catastrophe. PTSD was the most prevalent long-term psychiatric condition, followed by depressive disorders. Our results highlight the need to enhance preparedness and competence of health care professionals in detecting and managing the psychological sequelae of future comparable infectious disease outbreaks. PMID- 19555793 TI - Major depression and health-related quality of life in Parkinson's disease. AB - Depression is a common psychiatric condition in Parkinson's disease (PD), yet the burden of depression on health-related quality of life (HRQL) has not been clearly delineated in this patient population. OBJECTIVE: To evaluate the impact of depression and life stress on HRQL in the Canadian community dwelling population with PD. METHODS: A total of 259 respondents from the Canadian Community Health Survey (CCHS 1.1) with self-reported PD were interviewed. Measures included Health Utilities Index Mark 3 (HUI3), Composite International Diagnostic Interview Short Form for Major Depression and a single question regarding the amount of stress in their lives most days. Adjusted HUI3 scores were compared according to depression and life stress using ANCOVA models. RESULTS: Respondents without depression had overall HUI3 scores that were 0.29 units higher than respondents with depression [adjusted mean (95% CI) 0.49 (0.39 0.59) vs. 0.20 (0.03-0.37)]. The difference in overall HUI3 scores between respondents who reported high levels of stress as those who did not was 0.19 [adjusted mean (95% CI) 0.42 (0.29-0.55) vs. 0.23 (0.10-0.36)]. CONCLUSIONS: Substantial impact of depression and life stress, two modifiable factors, on HRQL is seen in PD. HRQL may be improved by clinical management of these nonmotor symptoms in PD. PMID- 19555792 TI - Typical somatic symptoms of pregnancy and their impact on a diagnosis of major depressive disorder. AB - OBJECTIVE: We sought to determine whether trimester of pregnancy influences the ability to diagnose major depressive disorder (MDD). METHOD: Eight hundred thirty eight subjects completed a Composite International Diagnostic Interview and the Edinburgh Postnatal Depression Scale (EPDS) before 17 weeks of pregnancy, at 26 30 weeks of pregnancy and at 4-12 weeks postpartum. Subjects responded to a checklist of MDD symptoms regardless of stem question endorsement. We compared rates of symptom expression by response (Y/N) to stem questions, and trimester, using logit analysis. Receiver operating characteristic curves determined optimal EPDS thresholds. RESULTS: Most symptoms from the DSM-IV checklist were endorsed significantly more often in the first compared to later trimesters (odds ratios ranged from 1.39 to 14.16 for the first vs. later trimesters), independent of response to depression stem questions or medication treatment. Despite this, stem positive and stem-negative groups differed significantly for 10 out of 13 symptoms (odds ratios, 2.29-6.89), independent of trimester. The EPDS had an optimal cutoff of 10 and showed acceptable predictive value. CONCLUSIONS: Pregnant women commonly experience somatic and other symptoms in this first trimester, but depressed women still differ from those who are not depressed. "Appetite increase," "oversleeping" and "increase in energy" (e.g., agitation) were uninformative with regard to an MDD diagnosis. PMID- 19555794 TI - Mortality in medical-surgical inpatients referred for psychiatric consultation. AB - OBJECTIVE: The purpose of this study is to calculate the 1-year mortality rate of medical-surgical inpatients referred for psychiatric consultation and to determine factors associated with mortality in this population. METHOD: We conducted a retrospective chart review of medical-surgical inpatients referred for psychiatric consultation (N=454). Patients who died within 1 year of psychiatric consultation were compared to age-matched patients from the same group who survived on demographic and clinical domains. RESULTS: Sixty-nine patients (15.2%) died within 1 year of referral for psychiatric consultation. Delirium was the only psychiatric diagnosis significantly associated with 1-year mortality [52.2% vs. 29.9%, P=.01; hazard ratio (HR), 1.7]. Cancer was the only medical condition associated with 1-year mortality (24.6% vs. 6.0%, P<.0001; HR, 3.03). Having both delirium and cancer shortened mean survival time significantly. Mortality risk was also associated with time duration between admission and psychiatric consultation. CONCLUSION: The 1-year mortality rate in this group of medical-surgical inpatients referred for psychiatric consultation was 15.2%. Mortality was independently associated with delirium and cancer. Mortality was also associated with greater length of time from admission to consultation, which raises a question about whether earlier psychiatry consultation may have a positive impact on survival. PMID- 19555795 TI - Coping, self-efficacy and psychiatric history in patients with both chronic widespread pain and chronic fatigue. AB - OBJECTIVE: To investigate the relationship of coping style and self-efficacy to functional impairment in a group of patients with both chronic widespread pain (CWP) and chronic fatigue, as well as the possible mediating role of psychiatric diagnosis. METHODS: We identified 138 consecutive clinic patients who met criteria for CWP and chronic fatigue. We collected demographic and clinical characteristics, as well as measures of emotion-focused and problem-focused coping styles, fatigue-related self-efficacy and self-reported general health. Psychiatric diagnoses were determined with a structured interview. Short Form-36 subscales of pain-related and fatigue-related functioning were the dependent variables in ordinal multiple regression analyses to identify the best-fit model for each. RESULTS: In the final model for pain, increased functional impairment was associated with increased emotion-focused coping as well as less education, lower general health scores and higher body mass index. Conversely, in the final model for fatigue, increased functional impairment was significantly associated with less emotion-focused coping, lower general health scores and lower self efficacy. CONCLUSIONS: The unexpected finding that emotion-focused coping was associated differently with chronic pain and fatigue among patients who experience both symptoms is discussed in the context of the research on the effects of self-efficacy and possible treatment approaches. PMID- 19555796 TI - Depression increases risk of incident myocardial infarction among Veterans Administration patients with rheumatoid arthritis. AB - OBJECTIVE: This study evaluates whether depression is a risk factor for incident myocardial infarction (MI) in Department of Veterans Affairs (VA) patients with rheumatoid arthritis (RA) between 30 and 79 years of age. METHODS: We used a retrospective cohort study of 15,634 patients with RA. Diagnoses and sociodemographic data were obtained from VA administrative and pharmacy databases between fiscal years 1999 and 2006. Entry into the cohort required 2 years of patient time with no evidence of cardiovascular disease. Cox proportional hazard models with time-dependent covariates were computed to determine whether RA patients with depression as compared to RA patients without depression were at increased risk for MI during the maximum 6-year follow-up period. RESULTS: Unadjusted analyses indicated depressed RA patients were 1.4 times more likely than nondepressed RA patients to have an MI during follow-up. These results remained significant (HR=1.4; 95% CI: 1.1-1.8) in the adjusted Cox proportional hazards model which included the effects of sociodemographics and known physical risks (e.g., diabetes) for MI. CONCLUSIONS: Depressed RA patients, without a history of cardiovascular disease, are 40% more likely to have a heart attack as compared to those without depression. These data demonstrate a rapid (within 6 years) transition to MI following onset of depression in RA patients. Increased monitoring of depression and heart disease status in this patient population may be warranted which in turn may result in longer duration of life. PMID- 19555797 TI - Psychiatric morbidity following electrical injury and its effects on cognitive functioning. AB - OBJECTIVE: This study examines the prevalence of psychiatric morbidity in a large sample of electrical injury (EI) patients in three phases of recovery and its effects on cognitive functioning. METHODS: Eight-six self-referred EI patients received psychiatric and neuropsychological evaluations. Descriptive statistics were conducted to examine the prevalence of psychiatric morbidity. Polytomous logistic regression was used to identify predictors of psychiatric diagnosis. Between-subjects analysis of variances (ANOVA) was conducted to examine the effects of psychiatric morbidity on cognitive functioning. RESULTS: Seventy-eight percent of subjects warranted a psychiatric diagnosis. Long-term patients compared to acute patients were more likely to be diagnosed with two diagnoses than not having any diagnosis (OR=14.30, 95% CI 1.40-38.71). Patients with two diagnoses performed worse than both patients with a single or no diagnosis on all cognitive outcome measures (P<.05). Voltage level, chronic pain and litigation status did not predict psychiatric morbidity. CONCLUSIONS: Psychiatric difficulties commonly emerge and persist following EI. EI patients with psychiatric conditions exhibited poorer cognitive performance as compared to EI patients with no post-injury psychiatric difficulties. Health care professionals need to devote careful attention to psychiatric and cognitive status when treating survivors of EI. PMID- 19555798 TI - First contact with psychiatric services: who leaves and who remains. AB - OBJECTIVE: The objective of this study was to obtain information on patients in their first contact with community mental health departments in the south of Italy, particularly on dropout patients leaving care without a previous agreement. METHOD: A 3-month cohort of 265 "first-contact" psychiatric patients assisted at four different community mental health centers (CMHCs) was examined and followed up at 6 months. RESULTS: The overall dropout rate after 6 months was 38.7%; it was higher for patients receiving pharmacological therapy alone and for patients seeking help on their own initiative, whereas physician-referred patients showed a lower dropout rate. More severely ill patients, as evaluated by physicians, showed dropout rates lower than those of patients "rating" themselves as severely ill. CONCLUSIONS: To lower dropout rates in CMHC settings, physicians should be provided with more concrete support in the patient selection and referral process. Greater focus should be placed on patients' motivational aspects and on their perceptions of their own symptom severity, as well as on the risks of dispensing "easy" solutions such as pharmacological therapy alone. PMID- 19555799 TI - Hypothermia and rhabdomyolysis following olanzapine injection in an adolescent with schizophreniform disorder. AB - Hypothermia and rhabdomyolysis are infrequent adverse effects of antipsychotic drugs. Here we report a case of an adolescent with schizophreniform disorder who developed both of them simultaneously after intramuscular injection of olanzapine. A 17-year-old male patient was hospitalized for treatment of psychotic symptoms, which persisted on risperidone 3 mg/day for 3 weeks. Then his antipsychotic drug was shifted to oral olanzapine 10 mg/day. The next day, he received intramuscular injection of olanzapine 5 mg and soon developed hypothermia, rhabdomyolysis, hypotension and bradycardia. These symptoms subsided gradually in the next 2 weeks after supportive treatment was given. Such adverse effects were not observed in the following 7 months. Possible pharmacological mechanisms were discussed. Physicians should be cautious about patients' clinical symptoms after giving olanzapine injection or rapid titration in dosage. PMID- 19555800 TI - Manifest psychosis in neurosyphilis. AB - INTRODUCTION: Recent surveys have assessed a remarkable increase in the prevalence of infectious syphilis. If left untreated, 30% of patients may develop tertiary syphilis, which can manifest as neurosyphilis. CASE REPORT: The authors present a case of an acute psychosis during and after pregnancy in a 37-year-old woman, which was identified as a manifestation of neurosyphilis after admission to a psychiatric ward. The positive screening test for syphilis provided the first hint of syphilis and gave direction for further diagnosis and specific treatment. Subsequently, the patient was treated with psychotropic medication concurrent with an adequate antibiotic treatment for neurosyphilis and was simultaneously psychologically monitored. An improvement of psychotic symptoms during antibiotic therapy was observed. CONCLUSION: This case emphasises that neurosyphilis still has to be considered in the differential diagnosis within the context of psychiatric conditions and diseases. Owing to current epidemiological data and difficulties in diagnosing syphilis, routine screening tests in the psychiatric field are necessary. PMID- 19555801 TI - Aripiprazole-induced hiccups: a case report. AB - Hiccups can arise from idiopathic, psychogenic and organic causes. The use of therapeutic drugs forms one of the important causes of hiccups. Although the exact pathophysiological processes involved have not yet been established, the neurotransmitters dopamine, serotonin and gamma amino butyric aid (GABA) have been documented to play a significant role in the generation of hiccups. We report a patient of organic bipolar affective disorder who developed hiccups with the atypical antipsychotic aripiprazole. The possible underlying neurotransmitter mechanisms, predisposing factors and clinical implications of this rare adverse event are discussed. PMID- 19555802 TI - Bilateral posterior vitreous detachment after electroconvulsive therapy. AB - In this case report, we describe a patient with an acute bilateral posterior detachment of the vitreous, which developed during electroconvulsive therapy (ECT) under general anesthesia. The patient developed symptoms of posterior vitreous detachment immediately after recovery from the eighth ECT session. The ophthalmologist confirmed the clinical diagnosis and ECT was stopped. We review the literature and discuss the ocular side effects of ECT. PMID- 19555803 TI - Akathisia and abnormal movements of the upper extremities with venlafaxine and methimazole. PMID- 19555804 TI - alpha/beta-Arteether-induced mania in a predisposed adolescent. AB - Artemisinin, by the name of Qinghaosu, has been used in China for the treatment of fever over the years. Recently, a number of artemisinin derivatives such as artesunate, artemether, dihydroartemisinin, and arteether have been developed and have found widespread clinical use because of their efficacy against resistant forms of all plasmodial species and a favorable side-effect profile. However, concerns have been expressed about the neurotoxic effects of artemisinin derivatives based on some animal and human studies. We present a case of alpha/beta-arteether-induced mania in an adolescent having a family history of chloroquine-induced psychosis to discuss the hereditary predispositions, possible mechanisms, management, and clinical implications of this rare adverse event. PMID- 19555805 TI - Gamma-hydroxybutyrate withdrawal syndrome: dangerous but not well-known. AB - Gamma-hydroxybutyrate (GHB) is an endogenous inhibitory neurotransmitter and anesthetic agent that is being abused as a 'club drug.' Withdrawal symptoms after cessation of GHB use are common and depend on the intensity of use. However, GHB withdrawal syndrome and delirium are unfamiliar to most psychiatrists, probably due to the fact that neither textbooks nor guidelines cover the subject. The GHB withdrawal syndrome may have a fulminant course that progresses to delirium. In those severe cases, admission to a general hospital and involvement of a psychiatrist become necessary. We present two cases of severe GHB withdrawal delirium, provide an overview of the literature and conclude with treatment recommendations. PMID- 19555806 TI - Should alternative diagnostic criteria be used to assess patients with cancer? A comment on Akechi et al. PMID- 19555808 TI - [The information about drugs on the Internet, a great step ahead]. PMID- 19555810 TI - Flavocoxid is as effective as naproxen for managing the signs and symptoms of osteoarthritis of the knee in humans: a short-term randomized, double-blind pilot study. AB - Flavocoxid (Limbrel), a proprietary mixture of flavonoid molecules (baicalin and catechin), was tested against a traditional nonsteroidal anti-inflammatory drug, naproxen, for the management of the signs and symptoms of moderate osteoarthritis (OA) in humans. Discomfort and global disease activity were used as the primary end points, and safety assessments were also taken for both treatments as a secondary endpoint. In this double-blind study, 103 subjects were randomly assigned to receive either flavocoxid [500 mg twice daily (BID)] or naproxen (500 mg BID) in a 1-month onset of action trial. Outcome measures included the short Western Ontario and McMaster University Osteoarthritis Index, subject Visual Analogue Scale for discomfort and global response, and investigator Visual Analogue Scale for global response and fecal occult blood. Both flavocoxid and naproxen showed significant reduction in the signs and symptoms of knee OA (P < or = .001). There were no statistically detectable differences between the flavocoxid and naproxen groups with respect to any of the outcome variables. Similarly, there were no statistically detectable differences between the groups with respect to any adverse event, although there was a trend toward a higher incidence of edema and nonspecific musculoskeletal discomfort in the naproxen group. In this short-term pilot study, flavocoxid was as effective as naproxen in controlling the signs and symptoms of OA of the knee and would present a safe and effective option for those individuals on traditional nonsteroidal anti inflammatory drugs or cyclooxygenase-2 inhibitors. A low incidence of adverse events was reported for both groups. PMID- 19555809 TI - L-Arginine enhances the triglyceride-lowering effect of simvastatin in patients with elevated plasma triglycerides. AB - We recently noticed a possible triglyceride-lowering effect during dietary supplementation with L-arginine. The major limitation of prior studies on L arginine, however, was that triglyceride levels were not the primary end point, and patients were not necessarily hypertriglyceridemic. Therefore, we conducted a 2-arm, randomized, double-blind study in 33 hypertriglyceridemic patients to investigate the hypothesis that oral L-arginine may lower serum triglyceride levels in hypertriglyceridemic patients on and off statins. The study consisted of a 6-week run-in phase, 6 weeks of treatment with L-arginine (n = 22, 1.5 g bid) or placebo (n = 11), and a 6-week extension period where simvastatin (20 mg qd) was added. All patients received dietary advice during each study visit. Routine and lipid laboratory parameters were determined in the local routine clinical laboratory. Treatment with L-arginine alone had no effects on serum lipids compared to placebo. The combination of L-arginine with simvastatin led to a significantly stronger reduction in triglycerides compared to placebo plus simvastatin (-140.5 +/- 149.2 mg/dL vs -56.1 +/- 85.0 mg/dL; P = .048). In addition, we found simvastatin-induced increases in aspartate transaminase and fibrinogen to be attenuated by L-arginine as compared to placebo. We conclude from our data that L-arginine enhances the effects of simvastatin on lipid metabolism, but it has no triglyceride-lowering effects when given alone. PMID- 19555811 TI - Cardiovascular risk factors in young, overweight, and obese European adults and associations with physical activity and omega-3 index. AB - Excess body fat is associated with increased cardiovascular disease (CVD) risk. The hypothesis of the study was that physical activity and omega-3 index, a marker of past long chain n-3 polyunsaturated fatty acids consumption, counteract the negative associations between fatness and CVD risk factors in young overweight and obese adults. A total of 324 subjects (20-40 years, body mass index [BMI], 27.5-32.5 kg/m(2), from Iceland, Spain, and Ireland) were investigated cross-sectionally. Dietary intake, anthropometric measurements, blood pressure, CVD risk factors, and fatty acids in erythrocyte membrane were analyzed. Information on physical activity was collected. Linear models were constructed to find out the associations of BMI, physical activity (quartiles), and omega-3 index with CVD risk factors. The most frequently increased risk factors were blood lipids (41.4%) and blood pressure (32.1%); fewer participants experienced disturbed glucose metabolism (11.8%). Body mass index was significantly associated with increased CVD risk factors (P = .001-.029), with the exception of total cholesterol, low-density lipoprotein, and high-density lipoprotein. The highest physical activity quartile had a lower fat mass (P = .005, at a given BMI), leptin (P = .008, in male participants only), and interleukin 6 (P = .021) but higher high-density lipoprotein (P = .020) than other quartiles; however, an approximate dose-response relationship could only be observed for leptin. The omega-3 index was not associated with lower low-density lipoprotein (P = .056), but docosahexaenoic acid in erythrocyte membrane was associated to it (P = .016). It is concluded that physical activity and docosahexaenoic acid diminish some of the negative health effects associated with overweight and obesity; however, body fatness remains the most important variable associated with increased CVD risk factors in young overweight and obese adults. PMID- 19555812 TI - Socioeconomic and gender differences in adolescents' nutritional status in urban Cameroon, Africa. AB - The aim of this study was to assess adolescents' nutritional status according to socioeconomic status (SES) and sex using anthropometry in urban Cameroon, Africa. Adolescent boys (n = 248) and girls (n = 333) 12 to 16 years old were recruited from randomly selected schools in a cross sectional study in Yaounde city and grouped according to SES. Weight, height, skinfold thickness, and circumferences were measured, and body mass index, waist/hip ratio, arm muscle, and arm fat areas were calculated. Stunting, underweight, and overweight were determined using international cutoff points. Adolescents with medium and high SES were less likely to be stunted than adolescents with low SES (odds ratio [OR], 0.40; P < .01). Prevalences of stunting (12%, 6%, and 5%) and underweight (3%, 4%, and 1%) were higher among the adolescents with low and medium SES than those with high SES. Overweight prevalence was high among the adolescents with low (8%), medium (11%), and high (9%) SES. The OR for overweight was higher among girls than boys (OR, 4.13; P < .001). Girls were less likely to be stunted and underweight than boys (OR, 0.29 [P < .001] and OR, 0.20 [P < .01], respectively). Prevalences of stunting (15% and 6%) and underweight (5% and 2%) were higher among boys than girls. Pubescent adolescents were less likely to be stunted than nonpubescent (OR, 0.53; P < .05). Adolescents with low and medium SES were more underweight and stunted than adolescents with high SES. Girls were more overweight, less stunted, and underweight than boys. PMID- 19555813 TI - Low 5-year stability of within-patient ion excretion and urine pH in fasting morning-urine specimens. AB - Most analyses of urine ion excretion and pH are based on 24-hour urine collections. However, fasting morning urine, collected in the morning after an initial void and a short fast, is easier to collect and may be sensitive to dietary intake. The objective of this study was to determine whether the within subject dietary acid load is stable by testing the levels of urine pH and ion excretion (ie, calcium, chloride, magnesium, phosphate, potassium, sodium, sulfate) in fasting morning urine specimens collected at a 5-year interval. Stable variables provide the best utility as potential risk factors. The subjects were 200 randomly selected adults (mean age = 61.5 +/- 11.1 years) from the 420 subjects who donated baseline and 5-year urine samples in the Canadian Multicentre Osteoporosis Study. The samples were collected in the morning after an initial void and a wait of 2 hours, while subjects maintained a fast from the evening before. The intraclass correlation coefficient was calculated to characterize the level of agreement between the baseline and 5-year urine samples as an indirect measure of diet stability. The stability of the within-subject urine measures over 5 years ranged from fair to moderate, and none were ranked as substantially stable. This fair-to-moderate stability of fasting morning urine measures of the diet acid load indicates a limited ability of a single sample of fasting morning urine to estimate subjects' actual long-term urine composition. PMID- 19555814 TI - Serum folate concentration, cognitive impairment, and DNA damage among elderly individuals in Malaysia. AB - The notion that dietary factors affect cognitive function and subsequently the risk of dementia has increased over the years from a global viewpoint. Because low folate intake has been described to impair cognitive function, we tested the hypothesis that low serum folate concentration is associated with cognitive impairment and an attenuated increase in DNA damage. We investigated the relationship between serum folate concentration, cognitive impairment, and DNA damage among elderly people attending health clinics in Klang Valley, an urban area in Malaysia. Two hundred thirty-two participants, composed of 115 men (49.6%) and 117 women (50.4%), were involved; none of the patients were diagnosed with neuropsychiatric problems, nor where they terminally ill. Sociodemography and health variables were assessed through face-to-face interview. Cognitive impairment review was conducted through an Elderly Cognitive Assessment Questionnaire. The estimation of dietary intake, serum folate concentration, and DNA damage was individually analyzed using validated Dietary History Questionnaires, immunoassay methods, and an Alkaline Comet Assay study (10 mL of peripheral venous blood), respectively. Results indicated that more men had cognitive impairment (33.0%) and DNA damage (27.0% for percentage DNA in tail, 22.6% tail moment) compared with women (25.6%, 15.4%, and 15.4%, respectively) (P < .05 for all parameters), recording an average folate deficiency value of 13.9% (0.2% higher than women). Multivariate binary logistic regression analysis outlined the association of cognitive impairment with older age (odds ratio [OR], 2.74; 95% confidence interval [CI], 1.35-5.57), smoking habits (OR, 5.12; 95% CI, 2.48-10.57), poor serum folate concentration (OR, 3.46; 95% CI, 1.26-9.52), and DNA damage (percentage DNA in tail) (OR, 13.70; 95% CI, 1.36-138.29). In conclusion, this study highlighted the important role of serum folate concentration for cognitive function and provided a concise picture regarding the elevated levels of oxidative DNA damage in peripheral lymphocytes. PMID- 19555815 TI - Ready-to-eat cereals are key sources of selected micronutrients among schoolchildren from public and private elementary schools in Quetzaltenango, Guatemala. AB - This cross-sectional dietary survey aimed to assess the consumption and relative nutrient contribution of ready-to-eat cereals (RTEC) among schoolchildren from 2 social classes in an urban center in the Guatemalan province of Quetzaltenango. A total of 449 24-hour dietary records were collected using a pictorial workbook registry method among third- and fourth-grade schoolchildren. The sample population was divided between low-income, public school attendants (n = 219) and students from higher-income private institutions (n = 230). We described the contribution of RTEC to estimated total energy; carbohydrates; protein; fat; vitamins A, C, and D; thiamin; riboflavin; folate; calcium; iron; and zinc over the 24-hour interval of registry. Approximately 41% of the subjects mentioned RTEC at least once in their 24-hour record; 93% of these at breakfast time. From the 7 RTEC varieties reported, 4 were presweetened. Estimated cereal consumption was significantly higher among private school participants (P < .001). The RTEC contributed 2.4% of estimated total energy across the whole sample and 3.6% of total carbohydrate; iron intake from RTEC was 21% of total, whereas less than 1% of calcium intake came from this source. Among the subgroup of cereal consumers, RTEC accounted for more than 40% of their daily recommendation intake for iron, vitamin C, thiamin, and riboflavin. The RTEC consumers had significantly higher intakes for all nutrients except carbohydrate and riboflavin as compared with nonconsumers. This study demonstrates the importance of RTEC as a key source of several selected micronutrients to schoolchildren's diet in this setting. PMID- 19555816 TI - Inadequate dietary magnesium intake increases atherosclerotic plaque development in rabbits. AB - Epidemiological studies have shown dietary magnesium (Mg) intake and serum Mg levels to be inversely correlated with the development of atherosclerosis. We hypothesized that low levels of Mg would promote atherosclerotic plaque development in rabbits. New Zealand white rabbits (4 months old, n = 22) were fed an atherogenic diet containing 0.12% (-Mg), 0.27% (control), or 0.43% (+Mg) Mg for 8 weeks. Blood samples were obtained at baseline, 2, 4, 6, and 8 weeks and were assayed for total cholesterol, high-density lipoprotein (HDL), non-HDL, triglycerides (TG), C-reactive protein, serum Mg, and erythrocyte Mg. Aortas from -Mg had significantly more plaque, with an intima thickness 42% greater than control and 36% greater than +Mg. Serum cholesterol levels rose over time, and at 8 weeks, -Mg had the highest and +Mg the lowest total and non-HDL cholesterol and TG levels, although these results did not reach significance. Over time, serum Mg levels increased, and erythrocyte Mg levels decreased. C-reactive protein significantly increased in all groups at 4 and 6 weeks but returned to baseline levels by 8 weeks. This study supports the hypothesis that inadequate intake of Mg results in an increase in atherosclerotic plaque development in rabbits. PMID- 19555822 TI - Movement disorders. Preface. PMID- 19555817 TI - Karaya root saponin exerts a hypocholesterolemic response in rats fed a high cholesterol diet. AB - Different sources of saponins are known to have hypocholesterolemic activity with varying degrees of efficacy. We hypothesize that karaya root saponin would efficiently reduce cholesterol. The aim of this study is to examine the comparative hypocholesterolemic effect of karaya root saponin in rats fed a high cholesterol diet. Sixty male Wister-Imamichi rats were divided into 5 groups of 12 rats each constituting of the following: control group, soybean saponin supplemented group, karaya root saponin-supplemented group, quillaja saponin supplemented group, and tea saponin-supplemented group. Compared with the control diet, both the karaya root- and quillaja saponin-supplemented diets significantly reduced (P < .05) serum cholesterol and atherogenic index. Karaya root saponin significantly increased the serum high-density lipoprotein cholesterol, high density lipoprotein cholesterol/cholesterol ratio, and fecal cholesterol concentrations (P < .05). The triacylglycerol concentration was significantly reduced only in the quillaja saponin-supplemented rats (P < .05). All the tea, soybean, karaya root, and quillaja saponins significantly reduced low-density lipoprotein cholesterol, and the greatest reduction was observed with karaya root saponin. Highest fecal bile acid concentration was found with quillaja saponin, whereas highest liver bile acid concentration was observed with karaya root saponin-supplemented rats (P < .05). These results collectively suggest that karaya root saponin can efficiently reduce serum cholesterol concentration in rats. PMID- 19555818 TI - Chemopreventive effect of raw and cooked lentils (Lens culinaris L) and soybeans (Glycine max) against azoxymethane-induced aberrant crypt foci. AB - Although lentils (Lens culinaris L) contain several bioactive compounds that have been linked to the prevention of cancer, the in vivo chemopreventive ability of lentils against chemically induced colorectal cancer has not been examined. Our present study examined the hypothesis that lentils could suppress the early carcinogenesis in vivo by virtue of their bioactive micro- and macroconstituents and that culinary thermal treatment could affect their chemopreventive potential. To accomplish this goal, we used raw whole lentils (RWL), raw split lentils (RSL), cooked whole lentils (CWL), and cooked split lentils (CSL). Raw soybeans (RSB; Glycine max) were used for the purpose of comparison with a well-studied chemopreventive agent. Sixty weanling Fischer 344 male rats, 4 to 5 weeks of age, were randomly assigned to 6 groups (10 rats/group): the control group (C) received AIN-93G diet, and treatment leguminous groups of RWL, CWL, RSL, CSL, and RSB received the treatment diets containing AIN-93G+5% of the above-mentioned legumes. After acclimatization for 1 week (at 5th to 6th week of age), all animals were put on the control and treatment diets separately for 5 weeks (from 6th to 11th week of age). At the end of the 5th week of feeding (end of 11th week of age), all rats received 2 subcutaneous injections of azoxymethane carcinogen at 15 mg/kg rat body weight per dose once a week for 2 consecutive weeks. After 17 weeks of the last azoxymethane injection (from 12th to 29th week of age), all rats were euthanized. Chemopreventive ability was assessed using colonic aberrant crypt foci and activity of hepatic glutathione-S-transferases. Significant reductions (P < .05) were found in total aberrant crypt foci number (mean +/- SEM) for RSB (27.33 +/- 4.32), CWL (33.44 +/- 4.56), and RSL (37.00 +/- 6.02) in comparison with the C group (58.33 +/- 8.46). Hepatic glutathione-S-transferases activities increased significantly (P < .05) in rats fed all treatment diets (from 51.38 +/- 3.66 to 67.94 +/- 2.01 micromol mg(-1) min(-1)) when compared with control (C) diet (26.13 +/- 1.01 micromol mg(-1) min(-1)). Our findings indicate that consumption of lentils might be protective against colon carcinogenesis and that hydrothermal treatment resulted in an improvement in the chemopreventive potential for the whole lentils. PMID- 19555823 TI - Etiology and pathogenesis of Parkinson disease. AB - The etiology of Parkinson disease (PD) is multifactorial and is likely to involve different causes in different patients. Several different genes have been identified as causes of familial PD, including alpha-synuclein gene mutations and multiplications, and mutations of parkin, PINK1, DJ1, and LRRK2. The biochemical consequences of these mutations have served to reinforce the relevance of the pathways to pathogenesis previously characterized, for example, mitochondrial dysfunction, oxidative stress, and protein misfolding and aggregation. The recognition that glucocerebrosidase mutations represent a significant risk factor for PD has focused attention on lysosomal function and autophagy as relevant to PD. Several environmental factors have also been shown to influence the risk for PD, although odds ratios remain relatively modest. Specific toxins can cause dopaminergic cell death in man and animals, but they probably have limited relevance to the etiology of PD. PMID- 19555824 TI - Medical treatment of Parkinson disease. AB - The cardinal characteristics of Parkinson disease (PD) include resting tremor, rigidity, and bradykinesia. Patients may also develop autonomic dysfunction, cognitive changes, psychiatric symptoms, sensory complaints, and sleep disturbances. The treatment of motor and non-motor symptoms of Parkinson disease is addressed in this article. PMID- 19555825 TI - Surgical treatment of movement disorders. AB - Surgical approaches are an important consideration in the management of many movement disorders, particularly for patients refractory to medications. In this article, we review the history, pathophysiology, risks and indications for surgical treatment. Summaries of case studies, case series and clinical trials performed using deep brain stimulation are provided for Parkinson's disease, dystonia, essential tremor and other movement disorders. PMID- 19555826 TI - Tremor: clinical features, pathophysiology, and treatment. AB - Tremor is not understood completely, and pharmacotherapy for all tremor disorders is inadequate. Fortunately, deep brain stimulation is effective for the most common and disabling tremor disorders. Our understanding of pathologic tremors has increased at an accelerating pace during the past 30 years, and this will hopefully lead to improved pharmacotherapy in the near future. PMID- 19555827 TI - Genetics and treatment of dystonia. AB - The torsion dystonias encompass a broad collection of etiologic subtypes, often divided into primary and secondary classes. Tremendous advances have been made in uncovering the genetic basis of dystonia, including discovery of a gene causing early onset primary torsion dystonia-a GAG deletion in exon 5 of the DYT1 gene that encodes torsinA. Although the exact function of torsinA remains elusive, evidence suggests aberrant localization and interaction of mutated protein; this may result in an abnormal response to stress or interference with cytoskeletal events and the development of neuronal brain pathways. Breakthroughs include the discovery of a genetic modifier that protects against clinical expression in DYT1 dystonia and the identification of the gene causing DYT6, THAP1. The authors review genetic etiologies and discuss phenotypes as well as counseling of patients regarding prognosis and progression of the disease. They also address pharmacologic and surgical treatment options for various forms of dystonia. PMID- 19555828 TI - Huntington disease and other choreas. AB - Chorea is defined as a syndrome characterized by brief, abrupt involuntary movements resulting from a continuous flow of random muscle contractions. There are genetic and non-genetic causes of chorea. The most common genetic cause of chorea is Huntington's disease (HD). Non-genetic forms of chorea include vascular choreas, auto-immune choreas, metabolic and toxic choreas, and drug-induced choreas. This chapter provides an overview of clinical features, pathogenesis and management of HD, other important genetic causes of chorea, Sydenham's chorea, other autoimmune choreas and vascular choreas. PMID- 19555829 TI - Tourette syndrome. AB - Tourette syndrome (TS) is a neuro-developmental disorder of childhood that is often associated with various psychiatric morbidities. Timely diagnosis and appropriate management can significantly impact psychosocial functioning. Morbidities may be a major source of disability, and may determine ultimate prognosis, although most children will experience significant improvement or resolution of symptoms by adulthood. Additional management considerations must be made in those with TS symptoms persisting into adulthood. The mainstay of therapy remains dopamine receptor blocking drugs, but new therapies are emerging. PMID- 19555830 TI - Pathophysiology and treatment of myoclonus. AB - Myoclonus is defined as sudden, brief, shock-like, involuntary movements caused by muscular contractions or inhibitions. Etiologic classification organizes the myoclonus disorders and provides major categories of clinical presentation. However, classifying myoclonus according to its source provides insight about its pathophysiology. The best strategy for symptomatic treatment is derived from defining the pathophysiology by way of source physiologic classification. PMID- 19555831 TI - Restless legs syndrome. AB - Restless legs syndrome (RLS) affects many people. General population prevalence surveys usually range from 1% to 12%, but most European ancestry studies suggest 10%. The development of validated rating scales and standardized diagnostic criteria have vastly improved the quality of RLS treatment trials. Although multiple medications have shown outstanding efficacy, all of them are felt to provide only symptomatic relief, rather than any "curative" effect. Dopamine agonists are clearly the best investigated and probably the most effective treatments for RLS. PMID- 19555833 TI - Peripherally induced movement disorders. AB - Peripherally induced movement disorders may be defined as involuntary or abnormal movements triggered by trauma to the cranial or peripheral nerves or roots. Although patients often recall some history of trauma before the onset of a movement disorder, determining the true relationship of the disorder to the earlier trauma is often difficult. The pathophysiology of these disorders is reviewed. PMID- 19555834 TI - Cardiac positron emission tomography. AB - Positron emission tomography (PET) is a powerful, quantitative imaging modality that has been used for decades to noninvasively investigate cardiovascular biology and physiology. Due to limited availability, methodologic complexity, and high costs, it has long been seen as a research tool and as a reference method for validation of other diagnostic approaches. This perception, fortunately, has changed significantly within recent years. Increasing diversity of therapeutic options for coronary artery disease, and increasing specificity of novel therapies for certain biologic pathways, has resulted in a clinical need for more accurate and specific diagnostic techniques. At the same time, the number of PET centers continues to grow, stimulated by PET's success in oncology. Methodologic advances as well as improved radiotracer availability have further contributed to more widespread use. Evidence for diagnostic and prognostic usefulness of myocardial perfusion and viability assessment by PET is increasing. Some studies suggest overall cost-effectiveness of the technique despite higher costs of a single study, because unnecessary follow-up procedures can be avoided. The advent of hybrid PET-computed tomography (CT), which enables integration of PET-derived biologic information with multislice CT-derived morphologic information, and the key role of PET in the development and translation of novel molecular-targeted imaging compounds, have further contributed to more widespread acceptance. Today, PET promises to play a leading diagnostic role on the pathway toward a future of high-powered, comprehensive, personalized, cardiovascular medicine. This review summarizes the state-of-the-art in current imaging methodology and clinical application, and outlines novel developments and future directions. PMID- 19555832 TI - Psychogenic movement disorders. AB - Psychogenic movement disorders (PMDs) represent a challenging dilemma for the treating neurologist. The terminology to classify this disorder is confusing and making the diagnosis is difficult. Once the diagnosis has been established, treatment options are limited, and the patient generally does not accept the diagnosis. PMID- 19555835 TI - Percutaneous coronary interventions in facilities without cardiac surgery on site: a report from the National Cardiovascular Data Registry (NCDR). PMID- 19555836 TI - Cardiac rehabilitation and survival in older coronary patients. AB - OBJECTIVES: This study assessed the effects of cardiac rehabilitation (CR) on survival in a large cohort of older coronary patients. BACKGROUND: Randomized controlled trials and meta-analyses have shown that CR improves survival. However, trial participants have been predominantly middle-aged, low- or moderate risk, white men. METHODS: The population consisted of 601,099 U.S. Medicare beneficiaries who were hospitalized for coronary conditions or cardiac revascularization procedures. One- to 5-year mortality rates were examined in CR users and nonusers using Medicare claims and 3 analytic techniques: propensity based matching, regression modeling, and instrumental variables. The first method used 70,040 matched pairs, and the other 2 techniques used the entire cohort. RESULTS: Only 12.2% of the cohort used CR, and those users averaged 24 sessions. Each technique showed significantly lower (p < 0.001) 1- to 5-year mortality rates in CR users than nonusers. Five-year mortality relative reductions were 34% in propensity-based matching, 26% from regression modeling, and 21% with instrumental variables. Mortality reductions extended to all demographic and clinical subgroups including patients with acute myocardial infarctions, those receiving revascularization procedures, and those with congestive heart failure. The CR users with 25 or more sessions were 19% relatively less likely to die over 5 years than matched CR users with 24 or fewer sessions (p < 0.001). CONCLUSIONS: Mortality rates were 21% to 34% lower in CR users than nonusers in this socioeconomically and clinically diverse, older population after extensive analyses to control for potential confounding. These results are of similar magnitude to those observed in published randomized controlled trials and meta analyses in younger, more selected populations. PMID- 19555837 TI - Analysis methods for observational studies: effects of cardiac rehabilitation on mortality of coronary patients. PMID- 19555838 TI - The pathophysiology of heart failure with normal ejection fraction: exercise echocardiography reveals complex abnormalities of both systolic and diastolic ventricular function involving torsion, untwist, and longitudinal motion. AB - OBJECTIVES: The purpose of this study was to test the hypothesis that in heart failure with normal ejection fraction (HFNEF) exercise limitation is due to combined systolic and diastolic abnormalities, particularly involving ventricular twist and deformation (strain) leading to reduced ventricular suction, delayed untwisting, and impaired early diastolic filling. BACKGROUND: A substantial proportion of patients with heart failure have a normal left ventricular ejection fraction. Currently the pathophysiology is considered to be due to abnormal myocardial stiffness and relaxation. METHODS: Patients with a diagnosis of HFNEF and proven cardiac limitation by cardiopulmonary exercise testing were studied by standard, tissue Doppler, and speckle tracking echocardiography at rest and on submaximal exercise. RESULTS: Fifty-six patients (39 women; mean age 72 +/- 7 years) with a clinical diagnosis of HFNEF and 27 age-matched healthy control subjects (19 women; mean age 70 +/- 7 years) had rest and exercise images of sufficient quality for analysis. At rest, systolic longitudinal and radial strain, systolic mitral annular velocities, and apical rotation were lower in patients, and all failed to rise normally on exercise. Systolic longitudinal functional reserve was also significantly lower in patients (p < 0.001). In diastole, patients had reduced and delayed untwisting, reduced left ventricular suction at rest and on exercise, and higher end-diastolic pressures. Mitral annular systolic and diastolic velocities, systolic left ventricular rotation, and early diastolic untwist on exercise correlated with peak VO(2)max. CONCLUSIONS: In HFNEF there are widespread abnormalities of both systolic and diastolic function that become more apparent on exercise. HFNEF is not an isolated disorder of diastole. PMID- 19555839 TI - The deconvolution of diastole. PMID- 19555840 TI - Computed tomographic angiography characteristics of atherosclerotic plaques subsequently resulting in acute coronary syndrome. AB - OBJECTIVES: In a computed tomographic (CT) angiography study, we identified the characteristics of atherosclerotic lesions that were associated with subsequent development of acute coronary syndrome (ACS). BACKGROUND: The CT characteristics of culprit lesions in ACS include positive vessel remodeling (PR) and low attenuation plaques (LAP). These 2 features have been observed in the lesions that have already resulted in ACS, but their prospective relation to ACS has not been previously described. METHODS: In 1,059 patients who underwent CT angiography, atherosclerotic lesions were analyzed for the presence of 2 features: PR and LAP. The remodeling index, and plaque and LAP areas and volumes were calculated. The plaque characteristics of lesions resulting in ACS during the follow-up of 27 +/- 10 months were evaluated. RESULTS: Of the 45 patients showing plaques with both PR and LAP (2-feature positive plaques), ACS developed in 10 (22.2%), compared with 1 (3.7%) of the 27 patients with plaques displaying either feature (1-feature positive plaques). In only 4 (0.5%) of the 820 patients with neither PR nor LAP (2-feature negative plaques) did ACS develop. None of the 167 patients with normal angiograms had acute coronary events (p < 0.001). ACS was independently predicted by PR and/or LAP (hazard ratio: 22.8, 95% confidence interval: 6.9 to 75.2, p < 0.001). Among 2- or 1-feature positive segments, those resulting in ACS demonstrated significantly larger remodeling index (126.7 +/- 3.9% vs. 113.4 +/- 1.6%, p = 0.003), plaque volume (134.9 +/- 14.1 mm(3) vs. 57.8 +/- 5.7 mm(3), p < 0.001), LAP volume (20.4 +/- 3.4 mm(3) vs. 1.1 +/- 1.4 mm(3), p < 0.001), and percent LAP/total plaque area (21.4 +/- 3.7 mm(2) vs. 7.7 +/- 1.5 mm(2), p = 0.001) compared with segments not resulting in ACS. CONCLUSIONS: The patients demonstrating positively remodeled coronary segments with low attenuation plaques on CT angiography were at a higher risk of ACS developing over time when compared with patients having lesions without these characteristics. PMID- 19555841 TI - Noninvasive detection of vulnerable coronary plaques: Locking the barn door before the horse is stolen. PMID- 19555842 TI - Incremental value of copeptin for rapid rule out of acute myocardial infarction. AB - OBJECTIVES: The purpose of this study was to examine the incremental value of copeptin for rapid rule out of acute myocardial infarction (AMI). BACKGROUND: The rapid and reliable exclusion of AMI is a major unmet clinical need. Copeptin, the C-terminal part of the vasopressin prohormone, as a marker of acute endogenous stress may be useful in this setting. METHODS: In 487 consecutive patients presenting to the emergency department with symptoms suggestive of AMI, we measured levels of copeptin at presentation, using a novel sandwich immunoluminometric assay in a blinded fashion. The final diagnosis was adjudicated by 2 independent cardiologists using all available data. RESULTS: The adjudicated final diagnosis was AMI in 81 patients (17%). Copeptin levels were significantly higher in AMI patients compared with those in patients having other diagnoses (median 20.8 pmol/l vs. 6.0 pmol/l, p < 0.001). The combination of troponin T and copeptin at initial presentation resulted in an area under the receiver-operating characteristic curve of 0.97 (95% confidence interval: 0.95 to 0.98), which was significantly higher than the 0.86 (95% confidence interval: 0.80 to 0.92) for troponin T alone (p < 0.001). A copeptin level <14 pmol/l in combination with a troponin T < or =0.01 microg/l correctly ruled out AMI with a sensitivity of 98.8% and a negative predictive value of 99.7%. CONCLUSIONS: The additional use of copeptin seems to allow a rapid and reliable rule out of AMI already at presentation and may thereby obviate the need for prolonged monitoring and serial blood sampling in the majority of patients. (Advantageous Predictors of Acute Coronary Syndromes Evaluation [APACE]; NCT00470587). PMID- 19555844 TI - Will 3.0-T make coronary magnetic resonance angiography competitive with computed tomography angiography? PMID- 19555843 TI - Contrast-enhanced whole-heart coronary magnetic resonance angiography at 3.0-T: a comparative study with X-ray angiography in a single center. AB - OBJECTIVES: The purpose of this study was to prospectively evaluate the diagnostic performance of 3.0-T contrast-enhanced whole-heart coronary magnetic resonance angiography (CMRA) in patients with suspected coronary artery disease (CAD). BACKGROUND: A slow-infusion, contrast-enhanced whole-heart CMRA approach has recently been developed at 3.0-T. The accuracy of this technique has not yet been determined among patients with suspected CAD. METHODS: The 3.0-T contrast enhanced whole-heart CMRA was performed in 69 consecutive patients. An electrocardiography-triggered, navigator-gated, inversion-recovery prepared, segmented gradient-echo sequence was used to acquire isotropic whole-heart CMRA with slow infusion of 0.2 mmol/kg gadobenate dimeglumine. The diagnostic accuracy of whole-heart CMRA in detecting significant stenoses (> or =50%) was evaluated using X-ray angiography as the reference. RESULTS: The CMRA examinations were successfully completed in 62 patients. Acquisition time of whole-heart CMRA procedure was 9.0 +/- 1.9 min. The 3.0-T whole-heart CMRA correctly identified significant CAD in 32 patients and correctly ruled out CAD in 23 patients. The sensitivity, specificity, and accuracy of whole-heart CMRA for detecting significant stenoses were 91.6% (87 of 95), 83.1% (570 of 686), and 84.1% (657 of 781), respectively, on a per-segment basis. These values were 94.1% (32 of 34), 82.1% (23 of 28), and 88.7% (55 of 62), respectively, on a per-patient basis. CONCLUSIONS: Contrast-enhanced whole-heart CMRA with 3.0-T allows for the accurate detection of coronary artery stenosis with high sensitivity and moderate specificity. PMID- 19555846 TI - Partial mechanical cardiac support: part of the solution or part of the problem? PMID- 19555845 TI - Proof of concept: hemodynamic response to long-term partial ventricular support with the synergy pocket micro-pump. AB - OBJECTIVES: The purpose of this study was to test the hemodynamic effects of partial ventricular support in patients with advanced heart failure. BACKGROUND: The use of current left ventricular assist devices (VADs) that provide full circulatory support is restricted to critically ill patients because of associated risks. Smaller, less-invasive devices could expand VAD use to a larger pool of less-sick patients but would pump less blood, providing only partial support. METHODS: The Synergy Pocket Micro-pump device (CircuLite, Inc., Saddle Brook, New Jersey) pumps approximately 3.0 l/min, is implanted (off pump) via a mini-thoracotomy, and is positioned in a right subclavicular subcutaneous pocket (like a pacemaker). The inflow cannula inserts into the left atrium; the outflow graft connects to the right subclavian artery. RESULTS: A total of 17 patients (14 men), age 53 +/- 9 years with ejection fraction 21 +/- 6%, mean arterial pressure 73 +/- 7 mm Hg, pulmonary capillary wedge pressure 29 +/- 6 mm Hg, and cardiac index 1.9 +/- 0.4 l/min/m(2) received an implant. Duration of support ranged from 6 to 213 (median 81) days. In addition to demonstration of significant acute hemodynamic improvements in the first day of support, 9 patients underwent follow-up right heart catheterization at 10.6 +/- 6 weeks. These patients showed significant increases in arterial pressure (67 +/- 8 mm Hg vs. 80 +/- 9 mm Hg, p = 0.01) and cardiac index (2.0 +/- 0.4 l/min/m(2) vs. 2.8 +/- 0.6 l/min/m(2), p = 0.01) with large reductions in pulmonary capillary wedge pressure (30 +/- 5 mm Hg vs. 18 +/- 5 mm Hg, p = 0.001). CONCLUSIONS: Partial support appears to interrupt the progressive hemodynamic deterioration typical of late-stage heart failure. If proven safe and durable, this device could be used in a relatively large population of patients with severe heart failure who are not sick enough to justify use of currently available full support VADs. (Safety and Performance Evaluation of CircuLite Synergy; NCT00878527). PMID- 19555847 TI - Cardiac manifestation of mucocutaneous lymph node syndrome (Kawasaki disease). PMID- 19555849 TI - Rethinking loading dose clopidogrel in light of increased bleeding complications in bypass patients. PMID- 19555850 TI - Aspirin for the masses. PMID- 19555852 TI - The 4th World Symposium on Pulmonary Hypertension. Introduction. PMID- 19555853 TI - Inflammation, growth factors, and pulmonary vascular remodeling. AB - Inflammatory processes are prominent in various types of human and experimental pulmonary hypertension (PH) and are increasingly recognized as major pathogenic components of pulmonary vascular remodeling. Macrophages, T and B lymphocytes, and dendritic cells are present in the vascular lesions of PH, whether in idiopathic pulmonary arterial hypertension (PAH) or PAH related to more classical forms of inflammatory syndromes such as connective tissue diseases, human immunodeficiency virus (HIV), or other viral etiologies. Similarly, the presence of circulating chemokines and cytokines, viral protein components (e.g., HIV-1 Nef), and increased expression of growth (such as vascular endothelial growth factor and platelet-derived growth factor) and transcriptional (e.g., nuclear factor of activated T cells or NFAT) factors in these patients are thought to contribute directly to further recruitment of inflammatory cells and proliferation of smooth muscle and endothelial cells. Other processes, such as mitochondrial and ion channel dysregulation, seem to convey a state of cellular resistance to apoptosis; this has recently emerged as a necessary event in the pathogenesis of pulmonary vascular remodeling. Thus, the recognition of complex inflammatory disturbances in the vascular remodeling process offers potential specific targets for therapy and has recently led to clinical trials investigating, for example, the use of tyrosine kinase inhibitors. This paper provides an overview of specific inflammatory pathways involving cells, chemokines and cytokines, cellular dysfunctions, growth factors, and viral proteins, highlighting their potential role in pulmonary vascular remodeling and the possibility of future targeted therapy. PMID- 19555854 TI - Future perspectives for the treatment of pulmonary arterial hypertension. AB - Over the past 2 decades, pulmonary arterial hypertension has evolved from a uniformly fatal condition to a chronic, manageable disease in many cases, the result of unparalleled development of new therapies and advances in early diagnosis. However, none of the currently available therapies is curative, so the search for new treatment strategies continues. With a deeper understanding of the genetics and the molecular mechanisms of pulmonary vascular disorders, we are now at the threshold of entering a new therapeutic era. Our working group addressed what can be expected in the near future. The topics span the understanding of genetic variations, novel antiproliferative treatments, the role of stem cells, the right ventricle as a therapeutic target, and strategies and challenges for the translation of novel experimental findings into clinical practice. PMID- 19555856 TI - Development and pathology of pulmonary hypertension. AB - The Development and Pathology working group was charged with reviewing the present knowledge, gaps in understanding, and areas for further studies in a broad range of themes. These themes in pulmonary vascular biology and pathobiology involved: 1) pulmonary vascular development; 2) pulmonary vascular disease accompanying fetal development and perinatal life; 3) properties of pulmonary vascular endothelial cells; 4) role of bone marrow cells in pulmonary vascular disease; 5) insights into pulmonary thromboembolic disease; 6) role of pathology in the assessment of pulmonary vascular disease; and 7) considerations of tissue banking for research in pulmonary hypertension. These important goals provide a blueprint for future research that may significantly impact our present and future understanding of pulmonary hypertension. PMID- 19555855 TI - Cellular and molecular basis of pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH) is caused by functional and structural changes in the pulmonary vasculature, leading to increased pulmonary vascular resistance. The process of pulmonary vascular remodeling is accompanied by endothelial dysfunction, activation of fibroblasts and smooth muscle cells, crosstalk between cells within the vascular wall, and recruitment of circulating progenitor cells. Recent findings have reestablished the role of chronic vasoconstriction in the remodeling process. Although the pathology of PAH in the lung is well known, this article is concerned with the cellular and molecular processes involved. In particular, we focus on the role of the Rho family guanosine triphosphatases in endothelial function and vasoconstriction. The crosstalk between endothelium and vascular smooth muscle is explored in the context of mutations in the bone morphogenetic protein type II receptor, alterations in angiopoietin-1/TIE2 signaling, and the serotonin pathway. We also review the role of voltage-gated K(+) channels and transient receptor potential channels in the regulation of cytosolic [Ca(2+)] and [K(+)], vasoconstriction, proliferation, and cell survival. We highlight the importance of the extracellular matrix as an active regulator of cell behavior and phenotype and evaluate the contribution of the glycoprotein tenascin-c as a key mediator of smooth muscle cell growth and survival. Finally, we discuss the origins of a cell type critical to the process of pulmonary vascular remodeling, the myofibroblast, and review the evidence supporting a contribution for the involvement of endothelial-mesenchymal transition and recruitment of circulating mesenchymal progenitor cells. PMID- 19555858 TI - Updated clinical classification of pulmonary hypertension. AB - The aim of a clinical classification of pulmonary hypertension (PH) is to group together different manifestations of disease sharing similarities in pathophysiologic mechanisms, clinical presentation, and therapeutic approaches. In 2003, during the 3rd World Symposium on Pulmonary Hypertension, the clinical classification of PH initially adopted in 1998 during the 2nd World Symposium was slightly modified. During the 4th World Symposium held in 2008, it was decided to maintain the general architecture and philosophy of the previous clinical classifications. The modifications adopted during this meeting principally concern Group 1, pulmonary arterial hypertension (PAH). This subgroup includes patients with PAH with a family history or patients with idiopathic PAH with germline mutations (e.g., bone morphogenetic protein receptor-2, activin receptor like kinase type 1, and endoglin). In the new classification, schistosomiasis and chronic hemolytic anemia appear as separate entities in the subgroup of PAH associated with identified diseases. Finally, it was decided to place pulmonary veno-occlusive disease and pulmonary capillary hemangiomatosis in a separate group, distinct from but very close to Group 1 (now called Group 1'). Thus, Group 1 of PAH is now more homogeneous. PMID- 19555857 TI - Genetics and genomics of pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH) is a rare disorder that may be hereditable (HPAH), idiopathic (IPAH), or associated with either drug-toxin exposures or other medical conditions. Familial cases have long been recognized and are usually due to mutations in the bone morphogenetic protein receptor type 2 gene (BMPR2), or, much less commonly, 2 other members of the transforming growth factor-beta superfamily, activin-like kinase-type 1 (ALK1) and endoglin (ENG), which are associated with hereditary hemorrhagic telangiectasia. In addition, approximately 20% of patients with IPAH carry mutations in BMPR2. We provide a summary of BMPR2 mutations associated with HPAH, most of which are unique to each family and are presumed to result in loss of function. We review the finding of missense variants and variants of unknown significance in BMPR2 in IPAH/HPAH, fenfluramine exposure, and PAH associated with congenital heart disease. Clinical testing for BMPR2 mutations is available and may be offered to HPAH and IPAH patients but should be preceded by genetic counseling, since lifetime penetrance is only 10% to 20%, and there are currently no known effective preventative measures. Identification of a familial mutation can be valuable in reproductive planning and identifying family members who are not mutation carriers and thus will not require lifelong surveillance. With advances in genomic technology and with international collaborative efforts, genome-wide association studies will be conducted to identify additional genes for HPAH, genetic modifiers for BMPR2 penetrance and genetic susceptibility to IPAH. In addition, collaborative studies of BMPR2 mutation carriers should enable identification of environmental modifiers, biomarkers for disease development and progression, and surrogate markers for efficacy end points in clinical drug development, thereby providing an invaluable resource for trials of PAH prevention. PMID- 19555859 TI - Diagnosis and assessment of pulmonary arterial hypertension. AB - The diagnosis and assessment of pulmonary arterial hypertension is a rapidly evolving area, with changes occurring in the definition of the disease, screening and diagnostic techniques, and staging and follow-up assessment. The definition of pulmonary hypertension has been simplified, and is now based on currently available evidence. There has been substantial progress in advancing the imaging techniques and biomarkers used to screen patients for the disease and to follow up their response to therapy. The importance of accurate assessment of right ventricular function in following up the clinical course and response to therapy is more fully appreciated. As new therapies are developed for pulmonary arterial hypertension, screening, prompt diagnosis, and accurate assessment of disease severity become increasingly important. A clear definition of pulmonary hypertension and the development of a rational approach to diagnostic assessment and follow-up using both conventional and new tools will be essential to deriving maximal benefit from our expanding therapeutic armamentarium. PMID- 19555860 TI - Interventional and surgical modalities of treatment in pulmonary hypertension. AB - Most patients with chronic thromboembolic pulmonary hypertension are operable, and pulmonary endarterectomy is the treatment of choice. Pulmonary endarterectomy should not be delayed for medical therapy, and risk stratification helps to define patients likely to achieve the best outcome. Inoperable patients should be referred for trials of medical agents. Atrial septostomy is promising but underutilized, although better ways of ensuring an adequate, lasting septostomy still need to be determined. Indications for the procedure are unchanged, and it should be considered more frequently. Bilateral sequential lung or heart-lung transplantation is an important option for selected patients, and potential candidates who are class IV or III but not improving should be referred early to a transplantation center. Currently, there is a need for right ventricular assist devices with flow characteristics suited to the circulation of patients with pulmonary arterial hypertension. Right ventricular synchronization therapy has not yet been tested. Novel shunts (e.g., Potts anastomosis) also hold promise. All surgery for pulmonary hypertension should be performed in centers with experience in these techniques. PMID- 19555861 TI - Updated evidence-based treatment algorithm in pulmonary arterial hypertension. AB - Uncontrolled and controlled clinical trials with different compounds and procedures are reviewed to define the risk-benefit profiles for therapeutic options in pulmonary arterial hypertension (PAH). A grading system for the level of evidence of treatments based on the controlled clinical trials performed with each compound is used to propose an evidence-based treatment algorithm. The algorithm includes drugs approved by regulatory agencies for the treatment of PAH and/or drugs available for other indications. The different treatments have been evaluated mainly in idiopathic PAH, heritable PAH, and in PAH associated with the scleroderma spectrum of diseases or with anorexigen use. Extrapolation of these recommendations to other PAH subgroups should be done with caution. Oral anticoagulation is proposed for most patients; diuretic treatment and supplemental oxygen are indicated in cases of fluid retention and hypoxemia, respectively. High doses of calcium-channel blockers are indicated only in the minority of patients who respond to acute vasoreactivity testing. Nonresponders to acute vasoreactivity testing or responders who remain in World Health Organization (WHO) functional class III, should be considered candidates for treatment with either an oral phosphodiesterase-5 inhibitor or an oral endothelin receptor antagonist. Continuous intravenous administration of epoprostenol remains the treatment of choice in WHO functional class IV patients. Combination therapy is recommended for patients treated with PAH monotherapy who remain in WHO functional class III. Atrial septostomy and lung transplantation are indicated for refractory patients or where medical treatment is unavailable. PMID- 19555862 TI - Diagnosis, assessment, and treatment of non-pulmonary arterial hypertension pulmonary hypertension. AB - The 4th World Symposium on Pulmonary Hypertension was the first international meeting to focus not only on pulmonary arterial hypertension (PAH) but also on the so-called non-PAH forms of pulmonary hypertension (PH). The term "non-PAH PH" summarizes those forms of PH that are found in groups 2 to 5 of the current classification of PH, that is, those forms associated with left heart disease, chronic lung disease, recurrent venous thromboembolism, and other diseases. Many of these forms of PH are much more common than PAH, but all of them have been less well studied, especially in terms of medical therapy. The working group on non-PAH PH focused mainly on 4 conditions: chronic obstructive lung disease, interstitial lung disease, chronic thromboembolic PH, and left heart disease. The medical literature regarding the role of PH in these diseases was reviewed, and recommendations regarding diagnosis and treatment of PH in these conditions are provided. Given the lack of robust clinical trials addressing PH in any of these conditions, it is important to conduct further studies to establish the role of medical therapy in non-PAH PH. PMID- 19555863 TI - End points and clinical trial design in pulmonary arterial hypertension. AB - New and emerging therapies might provide benefit in patients with pulmonary arterial hypertension. Their efficacy and safety will be compared with existing combination therapies in randomized clinical trials. Appropriate end points for these trials need to be identified: these will include exercise testing, the composite end point of time to clinical worsening, and hemodynamic markers, including advanced imaging modalities and biomarkers. Quality-of-life questionnaires are useful and important secondary end points; pulmonary arterial hypertension-specific questionnaires are currently being developed. Advantages and disadvantages of various trial designs, including placebo-controlled monotherapy or add-on trials, noninferiority studies, and withdrawal trials are also discussed. PMID- 19555864 TI - Preface: Hypertension. PMID- 19555865 TI - Epidemiology of hypertension in the elderly. AB - Hypertension is clearly associated with cardiovascular disease (CVD) and death. With age, the incidence of hypertension increases, making it imperative that we understand the pathophysiology and treatment of hypertension, especially in the elderly. Data regarding individuals older than 80 years are emerging, with more attention being given to patterns and treatment of hypertension in the elderly. Thus far, we have done a poor job with treating hypertension; this is due to multiple factors, including a reluctance of physicians to treat hypertension in the elderly because of concern of causing harm. In this article, the author's discuss the history and pathophysiology of hypertension, hypertension population studies, and hypertension treatment studies with a focus on the elderly. The author's findings both justify and encourage antihypertensive treatment in all hypertensive adults. PMID- 19555866 TI - Arterial compliance in the elderly: its effect on blood pressure measurement and cardiovascular outcomes. AB - The loss of arterial compliance as a result of the aging of human vasculature is thought to contribute to the age dependent rise in isolated systolic hypertension, and is an independent predictor of all cause mortality and cardiovascular outcomes. In this article, the author's we begin by providing a brief historical perspective of the study of the pulse wave. The author's then review the physiology of the normal arterial system, the effects of aging on the arterial system and the different measures of arterial stiffness. Finally, the author's review the different studies that look at arterial stiffness in the elderly, its impact on hypertension and cardiovascular outcome, and what is currently known on how to prevent vascular stiffness. PMID- 19555867 TI - General principles of hypertension management in the elderly. AB - The geriatric population is growing in number and along with it, the prevalence of hypertension (HTN). The elderly have a unique set of characteristics that must be taken into account when treating this condition. Not only is it widespread, but its consequences, mainly cardiovascular and cerebrovascular, are devastating. Because the elderly have multiple comorbid concomitant conditions, the practitioner must be cognizant of polypharmacy and resistant HTN and prescribe in a safe fashion conducive to compliance and efficacy. Treatment in even the oldest old is indicated. Function and quality of life should be the driving principles when managing the elderly, be they in the ambulatory or long-term care setting. PMID- 19555868 TI - Nonpharmacologic management of hypertension in the elderly. AB - Nonpharmacologic management of hypertension is all too often overlooked in the elderly. The Dietary Approaches to Stop Hypertension (DASH) diet, weight loss, physical activity, moderation of alcohol, and salt restriction, particularly when used in combination, are effective strategies to help control hypertension and reduce overall cardiovascular risk. Behavioral modification should form the cornerstone of hypertension treatment in the elderly. PMID- 19555869 TI - Polypharmacy in the elderly: focus on drug interactions and adherence in hypertension. AB - Polypharmacy is highly prevalent in the elderly due to an increased number of co morbid disease states that accompany aging. Hypertension is one common disease that can be challenging to treat in the elderly due to the body's physiologic changes, potential risks for side effects, medication interactions, and decreased medication adherence. A thorough medication assessment for each patient is essential when determining pharmacotherapeutic options in the elderly. PMID- 19555870 TI - Drug treatment of hypertension in older hypertensives. AB - There is strong evidence supporting the benefit of antihypertensive treatment in older patients. Blood pressure goal and drug selection in the elderly is similar to that in younger populations, but there are a few special considerations in these patients. A number of studies have been conducted to determine the drugs or drug classes most effective for reducing cardiovascular complications in older patients with hypertension. This article reviews the evidence for drug treatment in this population. PMID- 19555871 TI - Renin angiotensin system inhibition in the older person: a review. AB - The efficacy and safety of rennin angiotensin system (RAS) inhibition for lowering blood pressure in older populations has been demonstrated in a number of clinical trials. Whether a patient's age influences the overall ability of these drugs to lower blood pressure and protect against progress of cardiovascular and kidney disease has been the focus of few clinical trials. Herein, the author's review the mechanism of action of the renin angiotensin cascade and then discuss the clinical evidence surrounding the use of RAS-blocking drugs in the older population. PMID- 19555873 TI - Resistant hypertension in the elderly. AB - Resistant hypertension is more prevalent in the elderly population. Current data clearly shows the benefit of blood pressure control in older individuals. It is important to first differentiate pseudoresistance from true resistant and institute appropriate therapy. In those patients with resistant hypertension without an identifiable cause, non invasive measurement of hemodynamic profile is an important option to achieve meet blood pressure goals. PMID- 19555874 TI - Intraoperative MRI in functional neurosurgery. Preface. PMID- 19555876 TI - Intraoperative MRI: safety. AB - Intraoperative and interventional MR have opened a new chapter in neurosurgery bringing both new therapeutic opportunities and creating unique safety challenges for the MR operating room. The presence of a large magnetic field in the otherwise familiar environment of the operating room necessitates site-specific comprehensive policies for safety, staff training, infection control, and MR compatibility. Intraoperative MRI also creates unique MR image interpretation challenges that are of paramount significance for patient safety. These safety concerns are reviewed with particular reference to the nine years experience of the authors. PMID- 19555872 TI - Hypertension and cognitive function. AB - Cumulative evidence implicates hypertension in the pathogenesis of Alzheimer disease. Although it may not presently be possible to completely differentiate the effects of treatment and control of hypertension itself from those of the medication used to achieve such treatment goals, efforts directed at the treatment and control of hypertension can have significant public health impact. PMID- 19555875 TI - Origins of intraoperative MRI. AB - Neurosurgical diagnosis and intervention has evolved through improved neuroimaging, allowing better visualization of anatomy and pathology. This article discusses the various systems that have been designed over the last decade to meet the requirements of neurosurgical patients and opines on the potential future developments in the technology and application of intraoperative MRI. Because the greatest amount of experience with intraoperative MRI comes from its use in brain tumor resection, this article focuses on the origins of intraoperative MRI in relation to this field. PMID- 19555877 TI - Anesthesia in the intraoperative MRI environment. AB - Intraoperative MRI (iMRI) can be applied in several surgical settings. The incorporation of MRI technology into the operating room requires special considerations. The size and design of the operating room, including the equipment introduced into this setting, must be MR safe and allow adequate anesthesia monitoring and care. There are general restrictions and perils that may present in an operating room setting because of the MRI technology involving the monitoring equipment, anesthesia machine, and infusion devices. Incorporating the MRI technology into the operating room presents a new challenge in a transdisciplinary environment. The use of the iMRI technology has provided revolutionary tools for the new generation of medical practice. PMID- 19555878 TI - Intraoperative MRI with 1.5 Tesla in neurosurgery. AB - 1.5 Tesla High-field MRI has been successfully integrated into operating theatres. State-of-the-art microneurosurgical equipment and computer-assisted navigation are merged with the MR to form a comprehensive unit. The set-up of an intraoperative MRI solution is delineated with special regards to the workflow. PMID- 19555879 TI - Three-tesla high-field applications. AB - The authors believe that 3-T intraoperative MRI (iMRI) is likely to become the standard of care for a wide range of neurosurgical procedures. Although 3-T high field image acquisition does pose challenges, the advantages of this field strength, such as superior visualization of soft tissue and clear delineation of any residual tumor tissue, are clearly optimized using this equipment. Additionally, the use of 3-T high-field scanning offers functional options, such as brain activation studies and complex vascular imaging, that are unavailable with low- and midfield iMRI systems. The authors believe that the cost and effort necessary to acquire and establish a 3-T high-field iMRI program represent the natural progression for image-guided neurosurgery. PMID- 19555880 TI - Software requirements for interventional MR in restorative and functional neurosurgery. AB - Interventional MRI (iMRI) holds great promise for optimally guiding and monitoring restorative and functional neurosurgical procedures. This technology has already been used to guide ablative therapies and insert deep brain stimulation electrodes, and many future applications are envisioned. An optimized software interface is crucial for efficiently integrating the imaging data acquired during these procedures. MR systems are largely dedicated to image prescription and acquisition, whereas neuronavigation systems typically operate with previously acquired static data. An optimal software interface for iMRI requires fusion of many of the capabilities offered by these individual devices and further requires the development of tools to handle the integration and presentation of dynamically updated data. PMID- 19555881 TI - Devices for targeting the needle. AB - For decades the gold standard device for stereotactic procedures has been a frame based system that is rigidly attached to the head. More recently, frameless systems have been developed. More than 10 years after its introduction, the frameless system has not replaced its predecessor; most surgeons still prefer the frame-based system. Both systems rely on preoperative images that are registered to fiducial markers intraoperatively to meld image space to physical space. As a result, neither type of system is capable of providing real-time visualization of the brain target. A targeting device that works in conjunction with real-time intraoperative MRI scanners will address the primary limitation of today's targeting devices by allowing surgeons to work with real-time visualization of the brain target. PMID- 19555882 TI - Implantation of deep brain stimulator electrodes using interventional MRI. AB - The authors describe a method for placement of deep brain stimulator electrodes using interventional MRI in conjunction with a skull-mounted aiming device (Medtronic Nexframe). This approach adapts the procedure to a standard configuration 1.5-T diagnostic MRI scanner in a radiology suite. Preoperative imaging, device implantation, and postimplantation MRI are integrated into a single procedure performed under general anesthesia, providing real-time, high resolution magnetic resonance confirmation of electrode position. The method is conceptually simpler than the current standard technique for deep brain stimulator placement, as it eliminates the stereotactic frame, the subsequent requirement for registration of the brain in stereotactic space, physiologic testing, and the need for patient cooperation. With further technical refinement, the interventional MRI method should improve the accuracy, safety, and speed of deep brain stimulator electrode placement. PMID- 19555883 TI - Future applications: gene therapy. AB - Gene therapy for brain disorders is one of the most promising frontiers in the practice of restorative neurosurgery. There are significant experimental gene therapy initiatives underway that have led to currently active clinical trials using direct intracerebral delivery of viral vectors, and these treatments have been reported as safe and well tolerated. In the future, other clinical trials will likely use viral vectors to transfer genes that bestow on recipient tissue a desired enzymatic or neurotrophic activity relevant to the treatment of other neurodegenerative diseases, stroke, and traumatic brain injury. PMID- 19555884 TI - Future directions: use of interventional MRI for cell-based therapy of Parkinson disease. AB - Transplantation of neural cells for the treatment of neurologic disorders has garnered much attention and considerable enthusiasm from patients and physicians alike. Cell-based therapies have been proposed for a wide range of central nervous system pathologies ranging from stroke and trauma to demyelinating disorders and neurodegenerative diseases. Notably, cell transplantation for Parkinson disease (PD) has become even more attractive with the rapid advances in derivation of dopaminergic neurons from human embryonic stem cells. This article briefly reviews some of the relevant issues regarding the transplantation of cells for treatment of PD and hypothesizes how interventional MRI may be useful to optimize the surgical delivery of cells for PD and other central nervous system disorders. PMID- 19555885 TI - Uterine artery embolization under electroacupuncture for uterine leiomyomas. AB - PURPOSE: To evaluate whether electroacupuncture is a safe and effective alternative to pharmacologic sedation/analgesia in uterine artery embolization (UAE) for leiomyomas. MATERIALS AND METHODS: A nonrandomized prospective study was undertaken in 70 consecutive patients (mean age, 39.5 years) undergoing UAE with polyvinyl alcohol (PVA) particles between August 2006 and January 2007. Thirty-three patients chose to undergo UAE under electroacupuncture anesthesia (EAA; group A) and 37 were treated under local pharmacologic anesthesia (group B). Pain scores (rated from 0 to 10) in both groups were compared during and after the procedure. The outcome of UAE was evaluated at 6 months. RESULTS: Mean pain scores during embolization were 0.36 in group A and 0.84 in group B; scores after embolization and before discharge were 3.00 in group A and 4.49 in group B; and scores at discharge were 0.97 in group A and 2.11 in group B. These differences were statistically significant after embolization and at hospital discharge (P= .02 and P= .0001, respectively). All patients except one in each group were discharged from the hospital 4-8 hours after UAE; the two who remained longer had severe pain. There were no significant differences in clinical outcomes, nor in uterine and leiomyoma volumes, at discharge and at 6 months (P > 0.99 and P= .72, respectively). CONCLUSIONS: There was a statistically significant postembolization pain reduction in patients treated under EAA versus local pharmacologic anesthesia and no differences in UAE outcomes between groups at 6 months. PMID- 19555886 TI - Endobronchial dilation for the management of bronchial stenosis in patients after lung transplantation: effect of stent placement on survival. AB - PURPOSE: To compare the outcome of patients treated with balloon dilation and stent placement in the management of bronchial strictures after lung transplantation. MATERIALS AND METHODS: Forty-one lung recipients were treated with balloon dilation or stent placement between January 1997 and July 2005. Stent placement was reserved for cases of bronchoplasty technical failure or restenosis. Clinical files and results of pulmonary function tests and bronchoscopic evaluation were reviewed. Dyspnea and cough were defined according to the Breathlessness, Cough, and Sputum Scale. Patient survival and bronchial patency after bronchial intervention were estimated with the Kaplan-Meier method and Cox proportional hazards regression with analysis of stent implantation as a cofactor. RESULTS: Twenty-three of the 41 patients (56%) received a stent because of balloon dilation failure or stenosis recurrence. A total of 243 procedures were performed in 106 strictures (205 bronchoplasties and 38 stent insertions). At the first session, primary patency was higher in patients treated with stents (71%) than in those who underwent bronchoplasty (19%) (P = .037). Mean survival in patients with stents was longer than that in those who underwent bronchoplasty (82 vs 22 months, respectively), and stent insertion was associated with a 66% reduction in the risk of death (P < .02). Primary patency was 40 months for stented strictures versus 10 months for strictures treated with bronchoplasty (P < .02). Dyspnea and cough were improved after intervention (P < .001), and the forced expiratory volume in 1 second (FEV(1)) was ameliorated by 17% (P < .00003) at last follow-up. CONCLUSIONS: Clinical outcome and FEV(1) were improved after bronchoplasty and stent placement. Longer patient survival and bronchial patency were observed after stent insertion. PMID- 19555887 TI - Percutaneous drainage without sclerotherapy for benign ovarian cysts. AB - PURPOSE: To evaluate percutaneous short-term catheter drainage in the management of benign ovarian cysts in patients at increased surgical risk. MATERIALS AND METHODS: Thirty-eight patients with simple ovarian cysts were treated with drainage of fluid content by catheters until output stopped. All patients were poor candidates for surgery. All procedures were performed under ultrasonographic (US) control and local anesthesia. Cytologic examination was performed in all cases. The patients were followed up monthly with color Doppler US for 12 months. Outcome measure was the recurrence of a cyst. RESULTS: During the 12-month follow up period, 10 of 38 cysts recurred. Seven of the 10 cysts required further intervention, and three were followed up without intervention. Four of the seven patients who required further intervention underwent repeat transabdominal aspiration and three declined repeat aspiration and subsequently underwent surgery. After repeated aspirations, two of four cysts disappeared, one necessitated follow-up only, and one necessitated surgical intervention. Cyst volume (P = .009) and diameter (P = .001) were significantly larger in the cysts that recurred. No evidence of malignancy was reported in the cytologic examination in any patient. No patients developed malignancy during follow-up. No major complications were observed. The hospital stay was 1 day for all patients. The median duration of drainage in the groups with resolved and recurrent cysts was 1 day (interquartile range, 1-1) and 2 days (interquartile range, 1-3), respectively (P = .04). CONCLUSIONS: In patients considered poor candidates for open surgery or laparoscopy, percutaneous treatment of ovarian cysts with short term catheter drainage without sclerotherapy appears to be a safe and effective alternative, with low recurrence rates. PMID- 19555888 TI - Immunohistochemical characterization of specific inflammatory tissue reactions following embolization with four different spherical agents in the minipig kidney model. AB - PURPOSE: To evaluate the immunohistochemical inflammatory reaction after porcine renal embolization with the new spherical embolic agent Embozene and to compare it with other spherical embolic agents. MATERIALS AND METHODS: After superselective porcine renal embolization (40 pigs) with different sizes of embolic agents (Embozene, Embosphere, Bead Block, Contour SE), tissue arrays were obtained (size ranges, 40-120 microm, 100-300 microm, 500-700 microm, 700-900 microm). After immunostaining for CD subtyping (CD45 and CD68) and cytokines (C reactive protein [CRP] and interleukin-1 beta), a semiquantitative immunoreactivity score was calculated for each marker: intensity of staining was scored between 0 (negative) and 3 (intensive) and extent of staining between 0 and 4 (>80%), indicating the percentage of positive staining. The intensity score (0-3) was multiplied by the extent of staining score (0-4), resulting in a semiquantitative immunoreactivity score (0-12). RESULTS: Analysis of cellular expression profiles (ie, CD45, CD68) revealed a significantly higher inflammatory score 4 weeks after embolization with Embosphere 100-300 microm particles than after embolization with Embozene, Bead Block, and Contour SE. After 12 weeks, the Embosphere 100-300 microm score decreased. Analysis of CRP expression showed similar results, with a significantly higher score 4 weeks after embolization with Embosphere 100-300 microm. In the size class used most frequently for uterine artery emboliation (500-700 microm), all scores were low (<2.5) and there was no significant difference among particle types. CONCLUSIONS: Pronounced immunomarker expression was seen 4 weeks after embolization with small Embosphere particles. However, in general, modern spherical embolic agents cause a fairly low level of inflammatory reaction. In the present experimental setting, which is highly sensitive for specific tissue-to-agent reactivity, Embozene presented with low inflammatory results. PMID- 19555889 TI - The mouse arteriovenous fistula model. AB - PURPOSE: The first aim of the present study was to create a mouse carotid artery to-jugular vein arteriovenous (AV) fistula model. This model was used to test the hypothesis that there is increased gene expression of matrix metalloproteinase (MMP)-2 and MMP-9 at the venous stenosis. MATERIALS AND METHODS: Ten male FVB/NJ mice underwent the creation of an AV fistula between the left carotid artery and ipsilateral jugular vein, with the contralateral vessels serving as controls. Two mice died 1 day after surgery and the other eight were euthanized at day 28. Reverse transcriptase polymerase chain reaction was performed in five mice, with the grafted vein and control vein tissue used to determine the expression of MMP 2, MMP-9, TIMP-1, and TIMP-2. Immunohistochemical analysis of the grafted vein and control vein was performed in three mice. RESULTS: Venous stenosis formed at the outflow vein, characterized by a thickened neointima with cells staining positive for alpha-smooth muscle actin. There was increased expression of MMP-2, tissue inhibitor of metalloproteinase (TIMP)-1, and TIMP-2 by day 28 at the venous stenosis compared with control vein. CONCLUSIONS: A mouse carotid artery to-jugular vein AV fistula model was developed and used to demonstrate increased expression of several markers known to be associated with AV fistula stenosis. The model may be useful in investigating mechanisms responsible for AV fistula venous stenoses. PMID- 19555890 TI - In vitro study assessing the technical feasibility of using 3-F inner dilators for imaging. AB - PURPOSE: The 3-F inner dilators from three brands of micropuncture kits with slightly different designs were studied to identify the maximal flow rate of contrast medium infusion in hopes of identifying a safe range for the delivery of contrast media. MATERIALS AND METHODS: The inner dilators from three micropuncture kit manufacturers were tested. To assess maximum flow rates, each inner dilator was subjected to repeated contrast medium infusions at sequentially higher flow rates until it failed or reached maximal rate limits of the autoinjector. After a thorough inspection of each failed dilator, results for flow rates and pressures at failure were collected. RESULTS: All inner dilators were capable of achieving flow rates of at least 6 mL/sec at an average pressure of 258 psi. None of the inner dilators ruptured or resulted in a distal embolic event in our vessel model. Two dilators from the same manufacturer reached maximal rate limits of the autoinjector and did not fail. CONCLUSIONS: The use of the inner dilators from these micropuncture kits for nonselective angiography at rates at or less than 6 mL/sec appears technically feasible. Although it appears to offer angiographers and patients a quicker and perhaps safer alternative than conventional upsizing to catheters with larger lumens, additional experimental and clinical trials are necessary to fully endorse this in clinical practice. PMID- 19555892 TI - Fracture of a highly flexible nitinol stent after repeated bending of the knee joint during vigorous exercise. PMID- 19555891 TI - Ovarian artery embolization supplementing hypogastric-uterine artery embolization for control of severe postpartum hemorrhage: report of eight cases. AB - Although transcatheter uterine artery embolization has been proved to be a life saving technique in serious obstetrical hemorrhage, it does not always control bleeding. The causes of treatment failure may be multifactorial and vary in individual cases, but incomplete embolization of the blood supply from other sources may play an important role. Herein, the authors describe eight patients with massive postpartum hemorrhage that was not controlled with conventional hypogastric-uterine artery embolization. Their continued hemorrhage was due to an additional blood supply arising from ovarian arteries, which was confirmed with selective ovarian artery angiography and subsequently successfully embolized. PMID- 19555893 TI - Refractory bleeding in a TIPS patient due to an aberrant left gastric vein. PMID- 19555894 TI - Hypertensive crisis during radiofrequency ablation of the adrenal gland. PMID- 19555895 TI - Compulsory childhood vaccination. PMID- 19555896 TI - Protecting children with HIV against pneumococcal disease. PMID- 19555897 TI - Affordable medicines facility for malaria. PMID- 19555898 TI - Reacting to the emergence of swine-origin influenza A H1N1. PMID- 19555899 TI - It's a little bit more complicated than that. PMID- 19555900 TI - Virological monitoring and resistance to first-line highly active antiretroviral therapy in adults infected with HIV-1 treated under WHO guidelines: a systematic review and meta-analysis. AB - Antiretroviral-therapy rollout in resource-poor countries is often associated with limited, if any, HIV-RNA monitoring. The effect of variable monitoring on the emergence of resistance after therapy with commonly used drug combinations was assessed by systematic review of studies reporting resistance in patients infected with HIV with a CD4 count of fewer than 200 cells per muL treated with two nucleoside analogues (including a thymidine analogue) and a non-nucleoside reverse transcriptase inhibitor. 8376 patients from eight cohorts and two prospective studies were analysed. Resistance at virological failure to non nucleoside reverse transcriptase inhibitors at 48 weeks was 88.3% (95% CI 82.2 92.9) in infrequently monitored patients, compared with 61.0% (48.9-72.2) in frequently monitored patients (p<0.001). Lamivudine resistance was 80.5% (72.9 86.8) and 40.3% (29.1-52.2) in infrequently and frequently monitored patients, respectively (p<0.001); the prevalence of at least one thymidine analogue mutation was 27.8% (21.2-35.2) and 12.1% (5.9-21.4), respectively (p<0.001). Genotypic resistance at 48 weeks to lamivudine, nucleoside reverse transcriptase inhibitors (thymidine analogue mutations), and non-nucleoside reverse transcriptase inhibitors appears substantially higher in less frequently monitored patients. This Review highlights the need for cheap point-of-care viral load tests to identify early viral failures and limit the emergence of resistance. PMID- 19555901 TI - Mechanisms of avoidance of host immunity by Neisseria meningitidis and its effect on vaccine development. AB - Neisseria meningitidis remains an important cause of severe sepsis and meningitis worldwide. The bacterium is only found in human hosts, and so must continually coexist with the immune system. Consequently, N meningitidis uses multiple mechanisms to avoid being killed by antimicrobial proteins, phagocytes, and, crucially, the complement system. Much remains to be learnt about the strategies N meningitidis employs to evade aspects of immune killing, including mimicry of host molecules by bacterial structures such as capsule and lipopolysaccharide, which poses substantial problems for vaccine design. To date, available vaccines only protect individuals against subsets of meningococcal strains. However, two promising vaccines are currently being assessed in clinical trials and appear to offer good prospects for an effective means of protecting individuals against endemic serogroup B disease, which has proven to be a major challenge in vaccine research. PMID- 19555902 TI - Maternal and early onset neonatal bacterial sepsis: burden and strategies for prevention in sub-Saharan Africa. AB - Maternal and child health are high priorities for international development. Through a Review of published work, we show substantial gaps in current knowledge on incidence (cases per live births), aetiology, and risk factors for both maternal and early onset neonatal bacterial sepsis in sub-Saharan Africa. Although existing published data suggest that sepsis causes about 10% of all maternal deaths and 26% of neonatal deaths, these are likely to be considerable underestimates because of methodological limitations. Successful intervention strategies in resource-rich settings and early studies in sub-Saharan Africa suggest that the burden of maternal and early onset neonatal bacterial sepsis could be reduced through simple interventions, including antiseptic and antibiotic treatment. An effective way to expedite evidence to guide interventions and determine the incidence, aetiology, and risk factors for sepsis in sub-Saharan Africa would be through a multiarmed factorial intervention trial aimed at reducing both maternal and early onset neonatal bacterial sepsis in sub Saharan Africa. PMID- 19555903 TI - Bite-related and septic syndromes caused by cats and dogs. AB - Bite infections can contain a mix of anaerobes and aerobes from the patient's skin and the animal's oral cavity, including species of Pasteurella, Streptococcus, Fusobacterium, and Capnocytophaga. Domestic cat and dog bite wounds can produce substantial morbidity and often require specialised care techniques and specific antibiotic therapy. Bite wounds can be complicated by sepsis. Disseminated infections, particularly those caused by Capnocytophaga canimorsus and Pasteurella multocida, can lead to septic shock, meningitis, endocarditis, and other severe sequelae. An emerging syndrome in veterinary and human medicine is meticillin-resistant Staphylococcus aureus (MRSA) infections shared between pets and human handlers, particularly community-acquired MRSA disease involving the USA300 clone. Skin, soft-tissue, and surgical infections are the most common. MRSA-associated infections in pets are typically acquired from their owners and can potentially cycle between pets and their human acquaintances. PMID- 19555904 TI - Quinine monotherapy for treating uncomplicated malaria in the era of artemisinin based combination therapy: an appropriate public health policy? AB - Several African countries that have adopted artemisinin-based combination therapy (ACT) as first-line treatment of uncomplicated Plasmodium falciparum malaria also use quinine monotherapy as second-line therapy. This policy goes against WHO recommendations for combination therapy and could be considered an inappropriate public health policy. Adherence to a 7-day quinine treatment schedule is likely to be poor and may increase the risk of selecting resistant parasites. Furthermore, because quinine has limited post-treatment prophylaxis, it will not prevent, in areas of intense transmission, recurrent malaria infections, which can lead to additional morbidity, including anaemia. Therefore, ACTs and not quinine should be used as second-line treatment, because these are well tolerated, highly efficacious, and have the advantage of reducing gametocyte carriage and consequently malaria transmissibility, particularly in areas of less intense transmission. PMID- 19555905 TI - Syphilitic frosted branch angiitis with anterior uveitis in a patient with a new HIV diagnosis. PMID- 19555906 TI - A cold case of meningococcal meningitis. PMID- 19555907 TI - Midwifery and medical education: a decade of changes. PMID- 19555908 TI - Midwives as educators of medical students and residents: results of a national survey. AB - This descriptive study explores the roles and responsibilities of midwives involved in teaching medical students and residents. Surveys were received from 74 academic midwifery practices in the United States that are formally involved in medical education (66% response rate). These practices employ 547 midwives. Most of these midwives have >5 years of experience as clinicians (97%) and average >5 years of experience in medical education (73%). Academic midwifery practices teach multiple learners including obstetrics and gynecology residents (80%), family practice residents (60%), medical students in their core clerkship (93%), and midwifery students (83%). Midwives are often clinical preceptors during births of the midwifery practice's patients, but they assume a more formal academic faculty role with a resident or attending physician-shared case load of patients. Forty-two percent of midwifery practices care for their own patients along with the teaching and supervision of residents' patients. Roles and responsibilities of the midwifery practice members are explored, including the percentage of time spent in specific clinical and educational functions. Clinical experience and teaching expertise of midwives are valued in medical education. Multiple opportunities exist for collaboration between midwifery and medicine. PMID- 19555909 TI - A national survey of the midwifery director role in academic midwifery practices involved in medical education in the United States. AB - A survey to explore the role characteristics and key responsibilities of midwifery directors in academic practices involved in medical education in the United States was undertaken. Six key content areas were investigated: role responsibilities and characteristics, interaction with other medical divisions and committees, budgetary structure, interaction with learners, clinical schedules, and job satisfaction. A mail-based descriptive survey was distributed to 112 midwifery directors with a 56% response rate (N = 63). The results show a composite profile of academic midwifery practice directors involved in medical education that work primarily for departments of obstetrics and gynecology, are championed by the departmental chairperson, and have budgetary placement under this department. Collaboration with the residency director has not been fully realized, thereby limiting midwifery exposure and input regarding medical education, curricula changes, and access to key education committees. National changes in resident work hours had both a positive and negative impact on the director and overall midwifery practices. Job satisfaction documents both positive and challenging aspects to the director position, and most directors felt successfully integrated with physician colleagues. PMID- 19555910 TI - Documentation and billing for services provided by midwives teaching obstetrics and gynecology residents and medical students. AB - Certified nurse-midwives are teaching obstetrics and gynecology residents and medical students in major academic institutions across the United States. In these instances, the ability to appropriately document services rendered to support a billable service is paramount. This article explains the difference in requirements for midwives' documentation when working with residents compared with documentation required of an attending obstetrician-gynecologist. It also reviews the teaching physician guidelines developed by the Centers for Medicare and Medicaid Services (CMS) as well as current evaluation and management documentation requirements. Several examples of documentation are provided, as are suggestions for enhancement and simplification of the guidelines to include midwives. An important point to remember is that the CMS rules do not prohibit a certified nurse-midwife from teaching a resident. PMID- 19555911 TI - Models of organizational structure of midwifery practices located in institutions with residency programs. AB - Four models of organizational structure for midwifery practices that are located in academic institutions with residency programs are described: parallel models, coexistence models, fully integrated models, and blended models. Examples of each of these models are presented along with advantages and disadvantages and overall effect on resident education. PMID- 19555912 TI - Obstetric triage: models and trends in resident education by midwives. AB - Four models of resident education in obstetric triage with midwifery faculty consultants are presented. Common trends in the structure and function of these models are reviewed. The four models represent diverse settings where midwives serve as clinical teachers primarily for first-year obstetric residents and residents from other subspecialties. Each model supports a growing number of midwives working in the triage setting, functioning as both teacher and consultant for new residents. This expanded midwifery teaching role extends beyond labor assessment to include a wide range of common obstetric and gynecologic conditions in the triage setting. Additional advantages include the ability of the midwife to bill for triage services and to provide a safety net to decrease medical errors which, in a busy triage unit, occur most often during patient transfers. PMID- 19555913 TI - Innovative midwifery teaching for medical students and residents. AB - Many midwives are full-time faculty members of departments of obstetrics and gynecology in academic medical centers. As such, they contribute to the medical school's educational mission by teaching undergraduate and postgraduate medical students in both the clinical area and in didactic lectures. Midwives are also instrumental in the development of many innovative electives, workshops, and lectures, which provide students and residents with more exposure to the midwifery model of care. This article presents some of these innovations. PMID- 19555914 TI - Pioneering a universal curriculum: a look at six disciplines involved in women's health care. AB - This paper describes a multidisciplinary project of curriculum development and implementation at one university. Universal learning concepts for six health care disciplines in women's health, including nurse-midwifery, are being piloted in an online classroom environment. This multidisciplinary team approach to education distributes the work load, invites experts to share resources, and avoids the duplication of resources across the university departments and schools. This project provides midwifery faculty the opportunity to introduce a new educational paradigm and model the midwifery approach to holistic health care. PMID- 19555915 TI - The Collaboration for Maternal and Newborn Health: interprofessional maternity care education for medical, midwifery, and nursing students. AB - The Collaboration for Maternal and Newborn Health, a multidisciplinary group of maternity care providers from the University of British Columbia (UBC), received funding from Health Canada to develop interprofessional education programs for health care students. Medical, midwifery, and nursing students from UBC were invited to participate in the three programs described in this article. The Interprofessional Student Doula Support Program, a year-long program for 15 students, combines classroom learning about marginalized women with on-call doula support to attend births. The Interprofessional Normal Labour and Birth Workshop is a 5-hour event, comprised of lectures and hands-on stations about normal labour, birth, and the immediate postpartum period. The Maternity Care Club Hands on Night occurs twice a year, and students gather to practice at maternity care stations in a casual setting. A total of 467 participants over 3 years completed evaluations of their experiences. Students rate these programs very highly in terms of benefits of multidisciplinary collaboration. Providing students with opportunities to engage with other health care disciplines enhances interest in the professions of maternity care and the benefits of collaboration. PMID- 19555916 TI - Delayed clamping of the umbilical cord: a review with implications for practice. PMID- 19555919 TI - Examining ourselves: exploring assumptions about teaching pelvic examinations in midwifery education. PMID- 19555920 TI - Playing TETRIS for science counter-regulatory affective processing in a motivationally "hot" context. AB - We adapted the computer game TETRIS to investigate the process of affective motivational counter-regulation, that is, attentional biases for emotional stimuli that are in opposition to the momentary motivational focus. Counter regulation is seen as a mechanism which should prevent escalation and impulsivity, and it should help to avoid becoming "locked up" in affective motivational states. Accordingly, for a negative outcome focus condition (i.e., risk of losing a current high score), we hypothesized greater interference by positive distractors that were included in the game, whereas for a positive outcome focus (i.e., chance to improve one's current high score), we hypothesized greater interference by negative distractors. Supporting our hypotheses, we found the predicted interactions between distractor valence and type of outcome focus. PMID- 19555921 TI - Endothelial cell density after descemet membrane endothelial keratoplasty: 1- to 2-year follow-up. AB - PURPOSE: To evaluate donor endothelial cell density (ECD) after Descemet membrane endothelial keratoplasty (DMEK). DESIGN: Nonrandomized, prospective clinical study. METHODS: From a larger group of patients who underwent DMEK for Fuchs endothelial dystrophy or pseudophakic bullous keratopathy, complete ECD measurements were available of 26 patients with 6 and 12 months of follow-up, of whom 7 also had 24 months of follow-up. RESULTS: For the group with 24 months of follow-up, ECD averaged 2700 (+/- 260) cells/mm(2) before surgery, 2200 (+/- 460) cells/mm(2) at 6 months after surgery, 2050 (+/- 330) cells/mm(2) at 12 months after surgery, and 1780 (+/- 390) cells/mm(2) at 24 months after surgery. For the group with 12 months of follow-up, ECD averaged 2620 (+/- 210) cells/mm(2) before surgery, 1850 (+/- 540) cells/mm(2) at 6 months after surgery, and 1680 (+/- 550) cells/mm(2) at 12 months after surgery. In both groups, the ECD decreased significantly between the preoperative and 6-month measurement (P < .05). CONCLUSIONS: Similar to earlier endothelial keratoplasty techniques, DMEK may be associated with a decrease in donor ECD of approximately 25% in the early postoperative phase. PMID- 19555922 TI - SELDI-TOF-MS of gingival crevicular fluid--a methodological approach. AB - Human neutrophil peptides (HNPs) and the human cathelicidin LL-37 are antimicrobial peptides secreted by neutrophils, which play a crucial role in innate immune responses. The aim of this study was to establish a new method for ProteinChip arrays in combination with surface enhanced laser desorption/ionization (SELDI) technology and time-of-flight mass spectrometry to analyze gingival crevicular fluid (GCF) samples. To optimize experimental conditions, four different ProteinChip arrays (NP20; CM10, pH 4; CM10, pH 7; IMAC) along with corresponding binding buffers were tested. GCF samples were collected from patients showing healthy periodontal sites and sites with early signs of inflammation (gingivitis), but with no pocket depth greater than 4 mm. For GCF analysis, NP20 arrays and CM10 (pH 4) arrays showed specific and reproducible profiles in the range of 2.5-30.0 kDa. Donors that demonstrated significantly higher intensity peaks corresponding to the mass of LL-37 (p=0.01) also tended to show greater intensity peaks corresponding to the masses of HNP-1 and HNP-2 in samples from inflamed compared to healthy periodontal sites. The findings indicate that analysis of GCF samples by SELDI-TOF mass spectrometry is a useful approach to simultaneously analyze multiple markers, such as antimicrobial peptides, which may be beneficial for determination of new periodontal risk factors. PMID- 19555923 TI - Salivary levels of IL-1beta, IL-6, IL-8, and TNF-alpha in patients with burning mouth syndrome. AB - OBJECTIVE: To compare salivary IL-1beta, IL-6, IL-8, and TNF-alpha levels between patients with burning mouth syndrome (BMS) and controls. DESIGN: Forty female patients with BMS (mean age: 61.6+/-10.1 years) and 20 female control subjects (mean age: 65.1+/-9.0 years) were included in the study. Unstimulated (UWS) and stimulated whole saliva samples (SWS) were collected and their flow rates were determined. Salivary IL-1beta, IL-6, IL-8, and TNF-alpha levels and total protein concentration were also determined. Salivary transferrin level was determined to investigate the level of blood contamination in saliva samples. Gingival index of the subjects was also examined. Student's t-test, Pearson's correlation analysis, and analysis of covariance were used. RESULTS: No significant differences were found in the salivary levels of IL-1beta, IL-6, IL-8, and TNF-alpha in BMS patients compared with controls. Salivary flow rates and their total protein concentrations did not differ significantly between the groups. The levels of salivary cytokines and total protein concentration correlated significantly with the level of blood contamination in both UWS and SWS. CONCLUSION: There were no differences in the salivary levels of IL-1beta, IL-6, IL-8, and TNF-alpha in BMS patients compared with controls. Cytokine levels in whole saliva were affected mainly by the amount of blood contamination. PMID- 19555924 TI - Reproducible effects of subjectively assessed muscle fatigue on an inhibitory jaw reflex in humans. AB - OBJECTIVE: To evaluate the effects of exercise-induced, subjectively assessed muscle fatigue on an inhibitory jaw reflex, evoked by electrical stimulation of the upper lip. In addition, the reproducibility of these effects was assessed. DESIGN: Eight subjects participated in two experimental sessions that were two weeks apart. During each session, a baseline recording, a post-conditioning recording, and two recovery recordings were obtained. The post-conditioning recording was obtained immediately after provocation of jaw muscle fatigue by intense chewing. The endpoint of provocation was reached 30s after a subject had crossed the value '6' on a 10 cm long visual analogue scale. RESULTS: Subjectively assessed jaw muscle fatigue caused a decrease of about 50% in the size of the late inhibition in the post-conditioning recording (ANOVA: p=0.001; Bonferroni contrasts: p<0.05). Full recovery to baseline values was already achieved at the first recovery recording. No significant differences were found between both sessions (ANOVA, p=0.677). CONCLUSION: Exercise-induced, subjectively assessed jaw muscle fatigue causes a reproducible, transient suppression in the size of the late inhibitory jaw reflex wave. PMID- 19555925 TI - Chitosan membranes modified by contact with poly(acrylic acid). AB - In this work chitosan membranes modified by contact with poly(acrylic acid) (PAA) aqueous solution at two different temperatures (25 degrees C and 60 degrees C) were obtained. The pure chitosan (CS) membranes, as well as those treated with PAA (CSPAA_25 and CSPAA_60) were characterized by FTIR-ATR, water sorption capacity, thermal analysis (TG/DTG), and scanning electron microscopy (SEM). In addition, in vitro permeation experiments were carried out using metronidazol and sodium sulfamerazine aqueous solutions at 0.1% and 0.2% as model drugs. FTIR-ATR results showed the presence of absorption bands of NH3+ and COO(-) indicating the formation of a polyelectrolyte complex between chitosan and poly(acrylic acid). The results also indicated that PAA penetrates deeper into the membrane at higher temperature (60 degrees C), forming a thicker complex layer. Polyelectrolyte complex formation as well as the influence of treatment temperature was confirmed by lower hydrophilicity, higher thermal stability, and lower permeability of the treated membranes. The results show that the methodology used is a simple and very efficient way to drastically change some membrane properties, especially their permeability. PMID- 19555927 TI - [Why is it so difficult to integrate laparoscopic cholecystectomy as routine major ambulatory surgery?]. PMID- 19555926 TI - First total synthesis of 1,2-dipalmitoyl-3-(N-palmitoyl-6'-amino-6'-deoxy-alpha-D glucosyl)-sn-glycerol--a glycoglycerolipid of a marine alga with a high inhibitor activity against human Myt1-kinase. AB - The first total synthesis of 1,2-dipalmitoyl-3-(N-palmitoyl-6'-amino-6'-deoxy alpha-D-glucosyl)-sn-glycerol, a glycoglycerolipid isolated from a marine alga extract, is described. Starting from alpha-methylglucopyranoside the multistep strategy allows the stereoselective synthesis of the final compound using various protective group procedures as well as derivatization of partial molecule domains. The latter offers the development of lead structures for inhibitors of human Myt1-kinase. PMID- 19555928 TI - [The patient who could not wear her bikini. An amazing clinical presentation of sacral chordoma]. PMID- 19555929 TI - [Ileourethral fistula]. PMID- 19555930 TI - [Pericaecal hernias: a case presentation]. PMID- 19555931 TI - Statistics over features: EEG signals analysis. AB - This paper presented the usage of statistics over the set of the features representing the electroencephalogram (EEG) signals. Since classification is more accurate when the pattern is simplified through representation by important features, feature extraction and selection play an important role in classifying systems such as neural networks. Multilayer perceptron neural network (MLPNN) architectures were formulated and used as basis for detection of electroencephalographic changes. Three types of EEG signals (EEG signals recorded from healthy volunteers with eyes open, epilepsy patients in the epileptogenic zone during a seizure-free interval, and epilepsy patients during epileptic seizures) were classified. The selected Lyapunov exponents, wavelet coefficients and the power levels of power spectral density (PSD) values obtained by eigenvector methods of the EEG signals were used as inputs of the MLPNN trained with Levenberg-Marquardt algorithm. The classification results confirmed that the proposed MLPNN has potential in detecting the electroencephalographic changes. PMID- 19555932 TI - The role of noun syntax in spoken word production: evidence from aphasia. AB - We describe MH who presents with agrammatic aphasia and anomia, and who produces semantic errors in the absence of a central semantic impairment. This pattern of performance implies damage to syntactic processes operating between semantics and phonological output. Damage here may lead to lexical selection errors and a deficit in combining words to form phrases. We investigated MH's knowledge and processing of noun syntax in mass and count nouns. She produced more count nouns than mass nouns. She showed impaired knowledge of noun syntax in judgement tasks and production tasks, with mass noun syntax being more impaired than count. We interpret these results in terms of a two-stage model of lexical retrieval. We propose that syntactic information represented at the lemma level is activated even in bare noun production, and can be differentially impaired across noun categories. That same damage can lead to semantic errors in production. For MH limited syntactic options are available to support production, and these favour count noun production. The data provide a new account of output semantic errors. PMID- 19555933 TI - There are age-related changes in neural connectivity during the encoding of positive, but not negative, information. AB - INTRODUCTION: Older adults often show sustained attention toward positive information and an improved memory for positive events. Little is known about the neural changes that may underlie these effects, although recent research has suggested that older adults may show differential recruitment of prefrontal regions during the successful encoding of emotional information. In the present study, effective connectivity analyses examined the network of regions that college-age and older adults recruited during the encoding of positive and negative images. METHODS: Participants viewed positive and negative images while undergoing a functional magnetic resonance imaging (fMRI) scan. Structural equation modeling was used to compare young and older adults' connectivity among regions of the emotional memory network while they encoded negative or positive items. RESULTS: Aging did not impact the connectivity among regions engaged during the encoding of negative information, but age differences did arise during the encoding of positive information. Most notably, in older adults, the ventromedial prefrontal cortex and amygdala strongly influenced hippocampal activity during the encoding of positive information. By contrast, in young adults, a strong thalamic influence on hippocampal activity was evident during encoding. CONCLUSIONS: These findings suggest that older adults' "positivity effect" may arise from age-related changes in the interactions between affect processing regions and the hippocampus during the encoding of positive information. PMID- 19555935 TI - Prevalence, associations, and trends of biliary-tract candidiasis: a prospective observational study. AB - BACKGROUND: Biliary obstruction and cholangitis are common problems in gastroenterology. Infections of the biliary tract with Candida and other fungal species have increasingly been seen in the last few years. OBJECTIVE: To investigate the prevalence, associations, and trends of biliary-tract candidiasis. DESIGN: A prospective, observational, diagnostic study. SETTING: University Hospital, Muenster, Germany. PATIENTS: Consecutive patients undergoing ERCP for various indications. RESULTS: In 54 of 123 patients, we found Candida species in bile samples (44%). In only 7 patients, candidiasis was suspected on endoscopy before mycologic proof. Only 4 of these 7 patients were correctly diagnosed with biliary candidiasis by simple morphologic aspects. The fungus was mainly differentiated as Candida albicans or Candida glabrata and rarely as Candida parapsilosis, Candida tropicalis, or other subspecies. Immunosuppression for various reasons was significantly associated with bile-duct candidiasis (P < .02). No significant association was found between positive fungal cultures and prior endoscopic sphincterotomy (P = .0824) or prior ERCP (P = .1152). Biliary candidiasis was neither associated with positive fungal cultures of buccal smears (P = .0722) nor with positive findings in stool samples (P = .0860). LIMITATIONS: Highly selected patient population. Buccal smears and stool samples were not obtained from all patients. Contamination artifacts cannot totally be excluded with the ERCP procedure. CONCLUSIONS: Candida species very frequently can be detected in the bile. Positive fungal cultures of bile samples are not just contamination artifacts. This has to be taken into account when designing an anti infectious treatment for recurrent cholangitis or even more cholangiosepsis. Especially in immunosuppressed patients or recipients of long-term antibiotic therapy, physicians should screen for biliary-tract candidiasis during endoscopic examination. PMID- 19555934 TI - Blood levels of lead, cadmium, and mercury in the Korean population: results from the Second Korean National Human Exposure and Bio-monitoring Examination. AB - In Korea, there have been a number of efforts to measure levels of exposure to environmental pollutants among the population. This paper focuses on investigating the distribution of, extent of, and factors influencing the blood levels of lead, cadmium, and mercury in the Korean population, working from data obtained from the Second Korean National Human Exposure and Bio-monitoring Examination. To that end, blood metal concentrations were analyzed from a total of 2369 participants who were 18 years of age and older. The geometric mean concentrations and their 95% confidence intervals of metals in blood were found to be lead, 1.72 microg/dL (95% CI, 1.68-1.76); cadmium, 1.02 microg/L (95% CI, 1.00-1.05); and mercury, 3.80 microg/L (95% CI, 3.66-3.93). Regression analyses indicate that the levels of metals in the blood are mainly influenced by gender, age, and the education levels of the participants. Current smoking status is also found to be a significant factor for increasing both lead and cadmium levels. Although our study, as the first nationwide survey of exposure to environmental pollutants in Korea, has value on its own, it should be expanded and extended in order to provide information on environmental exposure pathways and to watch for changes in the level of exposure to environmental pollutants among the population. PMID- 19555936 TI - Administration of olive oil followed by a low volume of polyethylene glycol electrolyte lavage solution improves patient satisfaction with right-side colonic cleansing over administration of the conventional volume of polyethylene glycol electrolyte lavage solution for colonoscopy preparation. AB - BACKGROUND: Proper bowel cleansing before colonoscopy is essential for satisfactory evaluation of the colon. The required consumption of a large volume of salty-tasting liquid, 4 L of polyethylene glycol-electrolyte lavage solution (PEG-ELS), is the primary limitation to achieving this goal. OBJECTIVE: To achieve better patient satisfaction with efficient bowel cleansing, we compared the effects of the conventional volume (4 L) of PEG-ELS with those of a low volume (2 L) in combination with pretreatment using different laxatives, such as magnesium hydroxide (milk of magnesia) and olive oil. DESIGN: Randomized, controlled study. SETTING: A single research hospital. PATIENTS: Patients undergoing elective colonoscopy. INTERVENTIONS: A total of 120 patients were randomized to 1 of 3 different preparation regimens: 39 patients were prepared with a conventional volume (4 L) of PEG-ELS (Preparation [Prep] 1), and the remaining patients were prepared with a lower volume (2 L) of PEG-ELS and pretreatment with a laxative, either 15 g of magnesium hydroxide (40 patients, Prep 2) or 60 mL of olive oil (41 patients, Prep 3) 3 hours before PEG-ELS administration. MAIN OUTCOME MEASUREMENTS: The primary outcome was the efficacy of colonic cleansing on the left and right sides. Secondary outcomes were patient satisfaction and side effects. RESULTS: The olive oil regimen (Prep 3) resulted in significantly more adequate bowel cleansing of the right colon than administration of the conventional volume of PEG-ELS (Prep 1) and the magnesium hydroxide (Prep 2) regimen (97.6% vs 74.5% and 72.5%, respectively, P = .007). However, this difference was not observed in the left colon (91.5%, 85.5%, and 91.8% for Preps 1, 2, and 3, respectively, P = .776). When asked, 38 patients (95%) taking Prep 2, 35 patients (85.3%) taking Prep 3, and only 11 patients (28.2%) taking Prep 1 preferred the same preparation regimen if they required a future colonoscopy (P =.006), based on ease of use and taste. The side effects were comparable in each group. LIMITATIONS: The limitations of this study include the relatively small sample size, the single-center study design, and the use of a nonvalidated symptom scoring system. CONCLUSION: Pretreatment with olive oil before administration of a low volume of PEG-ELS enhances both patient satisfaction and the quality of right-side colonic cleansing over the administration of the conventional volume of PEG-ELS for colonoscopy preparation. Although the magnesium hydroxide regimen (Prep 2) was the preferred regimen in this study, its colonic cleansing effectiveness was not as great as those of the other regimens. Based on our results, the olive oil regimen is recommended, especially for patients who are not able to consume large volumes of liquids, such as the elderly, and those who are suspected of having subtle right-side colonic lesions. PMID- 19555937 TI - A rare complication during ERCP and sphincterotomy: placement of an endoscopic nasobiliary drainage tube in the portal vein. PMID- 19555938 TI - A randomized, controlled comparison of warm water infusion in lieu of air insufflation versus air insufflation for aiding colonoscopy insertion in sedated patients undergoing colorectal cancer screening and surveillance. AB - BACKGROUND: Pilot studies using a novel water method to perform screening colonoscopy allowed patients to complete colonoscopy without sedation medications and also significantly increased the cecal intubation success rate. OBJECTIVE: To perform a randomized, controlled trial comparing air insufflation (conventional method) and water infusion in lieu of air insufflation (study method) colonoscopy in minimally sedated patients. HYPOTHESIS: Compared with the conventional method, patients examined by the study method had lower pain scores and required less medication but had a similar cecal intubation rate and willingness to undergo colonoscopy in the future. SETTING: Outpatient colonoscopy in a single Veterans Affairs hospital. METHODS: After informed consent and standard bowel preparation, patients received premedications administered as 0.5-increments of fentanyl (25 microg) and 0.5-increments of Versed (midazolam) (1 mg) plus 50 mg of diphenhydramine. The conventional and the study methods for colonoscopy were implemented as previously described. Additional pain medications were administered at the patients' request. MAIN OUTCOME MEASUREMENTS: Increments of medications, pain scores, cecal intubation, and willingness to repeat colonoscopy. RESULTS: Increments of medications used before reaching the cecum (1.6 +/- 0.2 vs 2.4 +/- 0.2, P < .0027), total increments used (1.8 +/- 0.2 vs 2.5 +/- 0.2, P < .014), and the maximum pain scores (1.3 +/- 0.3 vs 4.1 +/- 0.6, P < .0002) were significantly lower with the water method. Cecal intubation rate (100%) and willingness to undergo a repeat colonoscopy (96%) were similar. LIMITATIONS: Single Veterans Affairs hospital, older male population. CONCLUSION: Water infusion in lieu of air insufflation is superior to air insufflation during colonoscopy in the minimally sedated patients (ClinicalTrials.gov Identifier NCT00785889). PMID- 19555939 TI - Removal of a Bravo 24-hour pH capsule with endoscopic scissors. PMID- 19555940 TI - Magnetic resonance imaging compatibility of endoclips. AB - BACKGROUND: Endoscopic clipping devices are now available for treatment of GI hemorrhage and microperforations. All commercially available endoclips are labeled as magnetic resonance imaging (MRI) incompatible. No data are available about the actual magnetic field strength at which endoclips are first deflected nor the clinical relevance of the magnetic fields on endoclips used in GI endoscopy. OBJECTIVE: To determine the compatibility of different endoclips with MRI. DESIGN: Prospective observational study. SETTING: Experiment on excised pig tissue in an MRI scanner. INTERVENTIONS: The physical deflection and strength of attraction of endoclips: Resolution Clip, TriClip, QuickClip, and Ethicon Endo surgery Clip were measured in different positions by using an MRI scanner at a field strength of 1.5 Tesla. Endoclips that demonstrated deflection were attached to a pig stomach and tested for detachment at a 1.5-Tesla MRI field strength. MAIN OUTCOME MEASUREMENTS: Physical deflection and detachment from pig stomach mucosa in an MRI scanner. RESULTS: All endoclips except the one made by Ethicon Endo-surgery demonstrated physical deflection under the tested conditions. The magnetic attraction was strongest for the Resolution Clip (0.7 gauss) compared with the TriClip (1.2 gauss) and the QuickClip (26.8 gauss). Only the Triclip demonstrated detachment from the pig gastric tissue under testing conditions. LIMITATIONS: A pig model and a small number of clips. CONCLUSIONS: The Ethicon Endo-surgery clip is compatible with MRI. All other clips showed deflection in a magnetic field, but only the TriClip demonstrated detachment from gastric tissue, and hence should be considered MRI incompatible. PMID- 19555941 TI - Pancreatitis is frequent among patients with side-branch intraductal papillary mucinous neoplasia diagnosed by EUS. AB - BACKGROUND: Because of greater recognition and improved imaging capabilities, intraductal papillary mucinous neoplasms (IPMNs) are being diagnosed with increasing frequency. IPMNs of the main pancreatic duct cause symptoms and lead to pancreatitis. Side-branch (SB) IPMNs are thought to cause symptoms less frequently, and their association with pancreatitis is not well defined. OBJECTIVE: Our purpose was to ascertain whether an association exists between SB IPMN and pancreatitis. DESIGN: Single-center, retrospective study. SETTING: Academic medical center. PATIENTS: A total of 305 patients underwent EUS examinations between October 2002 and October 2006 for pancreatic cystic lesions. MAIN OUTCOME MEASUREMENT: The main outcome measure was the frequency of acute or chronic pancreatitis that was not procedurally related. RESULTS: Thirty-two patients had SB-IPMNs, and 11 (34%) had pancreatitis. Three patients reported a single episode, and 8 patients reported having recurrent episodes of pancreatitis. Overall, 17 (53%) patients had symptoms possibly attributable to SB IPMN. Female sex (73% vs 38%) and multiple pancreatic lesions (54% vs 24%) were more commonly seen in those with pancreatitis, but were not statistically significant factors. Larger cyst size or cyst fluid marker levels did not appear associated with pancreatitis occurrence. EUS-FNA demonstrated communication with the pancreatic duct in 94% and thick, mucinous fluid in 84%. LIMITATIONS: Single center, retrospective study. CONCLUSIONS: Pancreatitis was frequently associated with the presence of SB-IPMNs in our referral practice. SB-IPMNs should be considered in the differential diagnosis of patients with recurrent pancreatitis with cystic lesions seen on imaging studies. EUS-FNA was the most useful modality in helping to differentiate SB-IPMNs from other lesions. PMID- 19555942 TI - Verrucous carcinoma of the esophagus. PMID- 19555943 TI - Assessment of colorectal adenomatous polyp size measured during pathological examination highlights the importance of accuracy. PMID- 19555945 TI - Mercury ingestion retrieved by colonoscopy. PMID- 19555944 TI - Treatment of weight regain after gastric bypass surgery when using a new endoscopic platform: initial experience and early outcomes (with video). AB - BACKGROUND: Some patients are thought to regain weight after gastric bypass surgery because of dilation of the gastrojejunal anastomosis (GJA) and/or gastric pouch. OBJECTIVE: To assess the technical success and safety of an endoscopic procedure, Revision Obesity Surgery Endoscopic (ROSE), using an endoscopic operating system for the placement of tissue anchors to reduce the diameter of the GJA and size of the gastric pouch. DESIGN: Prospective case series. SETTING: Brigham and Women's Hospital, Boston, Massachusetts. PATIENTS AND INTERVENTIONS: A total of 20 patients who had regained a mean of 13 kg after gastric bypass, with a dilated pouch and GJA on screening endoscopy, underwent the ROSE procedure. MAIN OUTCOME MEASUREMENTS: Technical success, reduction in size of the GJA and gastric pouch, and early complications. Early weight loss was also assessed. RESULTS: Technical success was achieved in 17 of 20 (85%) patients with an average reduction in stoma diameter of 16 mm (65% reduction) and an average reduction in pouch length of 2.5 cm (36% reduction). The mean weight loss in successful cases was 8.8 kg at 3 months. There were no major complications. LIMITATIONS: Single-center case series with a relatively small number of patients and short duration of follow-up, which was inadequate for assessing durable long term weight loss. CONCLUSION: The ROSE procedure is technically feasible and appears safe. Data from this series suggest that the ROSE procedure is effective in reducing not only the size of the GJA but also the gastric pouch and may provide an endoscopic alternative for weight regain in gastric bypass patients. PMID- 19555946 TI - The role of the endoscopist in a multidisciplinary obesity center. PMID- 19555947 TI - Role of double-balloon endoscopy in the diagnosis of small-bowel tumors: the first Japanese multicenter study. AB - BACKGROUND: The early diagnosis of small-bowel tumors (SBT) was a difficult task until the advent of capsule endoscopy (CE) and double-balloon endoscopy (DBE) allowed access to the disease site. However, although CE and DBE have greatly simplified the task, DBE studies have yet to provide sufficient data on the diagnosis and outcome of patients with SBTs. OBJECTIVE: To determine the efficacy of DBE examination in the detection and diagnosis of SBT. DESIGN: A retrospective analysis of cross-sectional case series. SETTING: Seven major medical centers in Japan. PATIENTS: The first 1035 consecutive DBE cases at these major Japanese centers since the introduction of DBE. MAIN OUTCOME MEASUREMENTS: The percentage of subjects with SBT and a diagnosis of SBT, their indications for DBE, and diagnostic and therapeutic DBE procedures carried out for SBT. RESULTS: SBTs were identified in 144 of 1035 subjects (13.9%) who underwent DBE between September 2000 and December 2005. For subjects with SBT, the most common indication for DBE was the suspected presence of a SBT (61/144 [42.4%]). For subjects without SBT, the most common indication was obscure GI bleeding (OGIB) (419/891 [47.0%]). Malignant lymphoma and GI stromal tumor (GIST) were the most frequent (31/144 [21.5%]) and the second-most frequent SBTs (27/144 [18.8%]), respectively, in this database. We also performed 85 biopsies and 45 therapeutic procedures for the evaluation and treatment of SBTs in 144 patients. Although complications were encountered in 5.3% of cases (14/266 sessions), none of these were life threatening in the present study. CONCLUSION: DBE proved a valuable tool for the detection and diagnosis of SBTs, especially when tumors were suspected. Biopsies and therapeutic procedures were also possible in most of these patients, which directed our management of the disease. PMID- 19555948 TI - Advanced pathology under squamous epithelium on initial EMR specimens in patients with Barrett's esophagus and high-grade dysplasia or intramucosal carcinoma: implications for surveillance and endotherapy management. AB - BACKGROUND: Prior studies report the presence of buried Barrett's epithelium under squamous mucosa after endoscopic ablative therapies for Barrett's esophagus (BE) with high-grade dysplasia (HGD) or intramucosal carcinoma (IMC). However, there still exists significant controversy about whether these glands represent a neoablative phenomenon or predate endoscopic therapy. OBJECTIVE: To determine the prevalence of buried BE underneath squamous epithelium on initial mucosectomy specimens for complete Barrett's eradication EMR (CBE-EMR) for BE with HGD or IMC. DESIGN: Retrospective double-blinded review. SETTING: A tertiary-care academic referral center. PATIENTS AND METHODS: Histopathology slides of all initial mucosectomy specimens for all patients who underwent CBE-EMR for BE with HGD or IMC at our center between August 2003 and February 2008 were reviewed retrospectively in a double-blinded fashion by 2 expert GI pathologists. None of the patients had undergone prior endoscopic ablative therapy for dysplastic BE. MAIN OUTCOME MEASUREMENTS: The prevalence of buried BE underneath squamous epithelium in initial mucosectomy specimens from CBE-EMR for BE with HGD or IMC. RESULTS: A total of 47 patients' initial mucosectomy slides were reviewed. The presence of Barrett's epithelium underneath the squamous resection margin (Z line) was identified in 13 of 47 patients (28%) at initial mucosectomy. The linear distance of the Barrett's epithelium from the resection's squamous margin ranged from 0.8 to 5.6 mm (mean 2.3 mm and median 1.9 mm). Histopathology revealed nondysplastic buried BE in 3 patients, HGD in 9 patients, and IMC in 1 patient. Thus, 10 of 13 patients (21% of 47 total) had buried glands with advanced pathology (HGD or IMC), whereas 3 of 13 (6% of 47 total) had specialized intestinal metaplasia without dysplasia. LIMITATIONS: A single-center, modest study population size. CONCLUSIONS: Our results revealed a significant prevalence of buried Barrett's epithelium with or without dysplasia under squamous mucosa (squamocolumnar junction) on initial mucosectomy specimens. Given the neoplastic potential of BE, the presence of these subsquamous BE glands may affect the extent and adequacy of mucosal resection margins. Based on these findings, surveillance biopsies and ablative therapy should extend to 1 cm proximal to the endoscopically determined squamocolumnar junction. PMID- 19555949 TI - Reversed LISS-DF in selected cases of complex proximal femur fractures. PMID- 19555950 TI - Compartment syndrome of the thigh: a systematic review. AB - INTRODUCTION: Thigh compartment syndrome is a surgical emergency with risk of high morbidity and mortality rates. The purpose of this study was to review the available evidence regarding the causes of thigh compartment syndrome, techniques of fasciotomy (specifically, one versus two incisions), methods of wound closure, and complications. METHODS: This institutional review board-exempt study was performed at a level-one trauma centre. PubMed and Medline OVID databases in the English language were searched for case series of two or more cases of compartment syndrome of the thigh. Cases were reviewed and analysed for causes of thigh compartment syndrome, number of fasciotomy incisions, methods of wound closure, and complications. RESULTS: A total of 9 papers met our criteria. All were retrospective case studies comprising a total of 89 patients. The most common cause was blunt trauma (90%). Motor vehicle accidents accounted for 36% of cases whilst motorcycle accidents were involved in 9%. Associated injuries included femur fractures in 48%, other limb fractures, renal, cardiovascular and head insults. Eighty-six percent of fasciotomies were performed through a single incision. Fifty-nine percent of fasciotomy wounds were closed by delayed primary closure, 26% had split-thickness skin grafts, and 15% had primary wound closure. Neurological deficits were the most common complications. CONCLUSION: There are limited data on thigh compartment syndrome with respect to cause, use of one versus two incisions for fasciotomy, methods of wound closure, and complication rates. Prospective studies are required to better define these variables in order to optimise the management of this problem. PMID- 19555951 TI - Placental and umbilical cord levels of neurokinin B and neurokinin B receptor in pre-eclampsia. PMID- 19555952 TI - Clinicopathologic analysis of uterine sarcomas from a single institution in the Canary Islands. AB - OBJECTIVE: To evaluate the clinicopathologic data and prognostic factors for patients with uterine sarcomas treated at a single institution, with special emphasis on malignant mixed mullerian tumors (MMMT). METHODS: Medical and anatomic pathology records were reviewed. Survival rates were analyzed using the Kaplan-Meier method. RESULTS: The study included 89 patients: 48.4% with MMMT; 22.4% with leiomyosarcomas; 20.2% with endometrial stromal sarcomas; and 9% with adenosarcomas. FIGO stages I, II, III, and IV were identified in 57.3%, 9.0%, 22.5%, and 7.8% of patients respectively. Event-free survival rates after 2, 5, and 10 years were 70%, 61%, and 55% respectively, with a median time of 90 months (95% CI, 41-140 months). Overall survival rates after 2, 5, and 10 years were 50%, 45%, and 39% respectively, with a median time of 43 months (95% CI, 3-83 months). Multivariate analysis showed that stage, histology, tumor size, and parity had an independent influence on overall survival. CONCLUSIONS: MMMT are the most aggressive tumors and their behavior strongly resembles that of high grade endometrial adenocarcinoma. Prognostic factors affecting survival were stage, histology, tumor size, and parity. PMID- 19555953 TI - Total laparoscopic radical parametrectomy, partial colpectomy, and pelvic lymphadenectomy in patients with occult cervical cancer. AB - OBJECTIVE: To assess total laparoscopic radical parametrectomy (TLRP) with pelvic lymphadenectomy and partial colpectomy as a safe and feasible treatment option for patients with occult cervical cancer. METHODS: Twelve patients with occult invasive cervical cancer underwent TLRP after prior extrafascial hysterectomy. RESULTS: No intraoperative complications occurred. Two patients experienced postoperative complications: an iliac lymphocyst with pyelectasis, and a vaginal evisceration that occurred during sexual intercourse. Nine patients required no further treatment. One patient with residual disease received brachytherapy as adjuvant treatment. Two patients with positive nodes not detected at preoperative work-up received adjuvant concomitant radiochemotherapy. CONCLUSION: TLRP with pelvic lymphadenectomy is a safe and feasible treatment in patients with occult invasive cervical cancer discovered after extrafascial hysterectomy. PMID- 19555954 TI - A comparison of novice and expert nurses' cue collection during clinical decision making: verbal protocol analysis. AB - BACKGROUND: The type of cues used during clinical decision-making contexts is not well understood. Further, there are conflicting findings in relation to how novice and expert nurses use cues. OBJECTIVE: The aim of this study was to determine if there were differences between novice and expert nurses in the range and type of cues selected as well as how cues were clustered together when making clinical decisions while caring for post-operative patients in an Intensive Care Unit. METHOD: The sample consisted of four novice and four expert nurses caring for patients post Abdominal Aortic Aneurysm surgery in an Intensive Care Unit. Data were collected using a think aloud (TA) process while participants cared for patients, followed by retrospective interviewing, to generate verbal protocols. The verbal protocols were analysed using content analysis to examine various aspects of decision-making, including number and type of cues used and cue clustering. The decision tasks attended in the real world of practice were described in detail to illuminate the use of cues in context. RESULTS: Expert nurses collected a wider range of cues than novice nurses, almost twice as many different cues. The expert nurses also clustered more cues together to identify patient status when making decisions. Expert nurses were more proactive in collecting relevant cues and anticipating problems that may help identify patient problems. CONCLUSIONS: In the real world of practice expert nurses collect a broader range of cues to assess patient status than novice nurses. This differs to expert nurses cue collection in simulations where expert nurses may select only those cues that are necessary for the identified problem. This difference, if identified in other studies, may have important implications for nursing research and education. PMID- 19555956 TI - [Laparoscopic surgery for colon cancer: a critical reading of the randomized trials of survival]. PMID- 19555955 TI - The driving and restraining forces that promote and impede the implementation of individualised nursing care: a literature review. AB - BACKGROUND: Despite substantial attention devoted to the development of individualised care in recent years, there is a lack of coherent information and empirical research assessing the driving and restraining forces that promote and impede its implementation. OBJECTIVES: The aim of this integrative literature review is to describe the driving and restraining forces for the implementation of individualised nursing care from the nurse's point of view. This information is useful for the development of clinical nursing care and in identifying areas for future research. DESIGN: An integrative literature review. DATA SOURCES: An integrative analysis of empirical studies and reviews derived from the MEDLINE, CINAHL databases and EMB Reviews - The Cochrane Database of Systematic Reviews (individual* care & nurse, from earliest through April 2008) was conducted focusing on studies which used nurses as informants. The final sample of 43 articles published in English focusing on the individualised care of adult patients from the nurse's point of view was retrieved after a two-stage process. REVIEW METHODS: The integrative analysis of the studies included three steps. Firstly, the full texts of the final eligible studies were read. Secondly, the driving and restraining forces, that respectively promote or impede the delivery of individualised care were identified and listed in a working sheet. A total of nine categories were identified. Thirdly, a working sheet was completed summarising the information found in the studies reviewed and listing the authors. RESULTS: Nine categories describing both driving and restraining forces for individualised nursing care were identified: (1) nurse's personal characteristics, (2) skills enhancement, (3) ethical issues, (4) nursing care delivery and interventions, (5) patient characteristics, (6) organisation of work, (7) staffing, (8) team work and group dynamics, and (9) leadership and management. CONCLUSIONS: A body of knowledge was identified for future research. The results inform both clinical practice and education and promote better use of the nursing work force in order to provide individualised care for patients and maximise good patient outcomes. PMID- 19555957 TI - Determination of dimethyl fumarate in desiccant and mouldproof agents using ultrasound-assisted extraction gas chromatography with electron-capture detection. AB - A fast, simple, low cost, and high throughput method has been developed for the determination of dimethyl fumarate (DMF) in desiccant and anti-mould agents employed for protecting clothes, footwear and accessories from humidity and mould. The procedure is based on ultrasound-assisted extraction followed by GC microECD analysis. The method was conveniently optimized, and the analyte was efficiently extracted from the samples in only 5 min using such a low volume of solvent (ethyl acetate) as 1 mL. Simultaneous extractions could be carried out in also 5 min without loosing efficiency. The method has been validated showing good linearity and selectivity. Precision was satisfactory with RSD of 4-5%. Recovery was evaluated in two real samples at different DMF concentration levels and was >90% in all cases. Limit of detection and quantification were at the ng g(-1) level. The proposed procedure was applied to the determination of DMF in several desiccant and anti-mould samples. Although most of them were labelled as "silica gel" all the samples tested with the exception of three presented important amounts of DMF, many of them at the high microg g(-1) level. The presence and the high concentrations of this allergenic fungicide in desiccant and anti-mould sachets employed in many consumer products, such as clothes and footwear, should be a matter of concern. PMID- 19555958 TI - An evaluation of microwave-assisted derivatization procedures using hyphenated mass spectrometric techniques. AB - The potential of microwave-assisted derivatization techniques in systematic toxicological analysis using gas chromatography coupled with mass spectrometry (GC-MS) was evaluated. Special emphasis was placed on the use of dedicated microwave reactors incorporating online temperature and pressure control. The use of such equipment allowed a detailed analysis of several microwave-assisted derivatization protocols comparing the efficiency of microwave and conventional heating methods utilizing a combination of GC-MS and liquid chromatography coupled with mass detection (LC-MS and LC-MS/MS) techniques. These studies revealed that for standard derivatization protocols such as acetylation (exemplified for codeine and morphine), pentafluoropropionylation (for 6 monoacetylmorphine) and trimethylsilylation (for Delta9-tetrahydrocannabinol) a reaction time of 5 min at 100 degrees C in a microwave reactor was sufficient to allow for an effective derivatization. Control experiments using standard operating procedures (30 min at 60 degrees C conventional heating) indicated that the faster derivatization under microwave irradiation is a consequence of the higher reaction temperatures that can rapidly be attained in a sealed vessel and the more efficient heat transfer to the reaction mixture applying direct in core microwave dielectric heating. The results suggest that microwave derivatization procedures can significantly reduce the overall analysis time and increase sample throughput for GC-MS-based analytical methods. PMID- 19555959 TI - Characteristics of BPA removal from water by PACl-Al13 in coagulation process. AB - This paper discussed the coagulation characteristics of BPA with polyaluminum chloride (PACl-Al(13)) as coagulant, examined the impact of coagulation pH, PACl Al(13) dosage, TOC (total organic carbon) and turbidity on BPA removal, and analyzed the possible dominant mechanisms in water coagulation process. Formation and performance of flocs during coagulation processes were monitored using photometric dispersion analyzer (PDA). When the concentration of humic acid matters and turbidity was low in the solution, the experimental results showed that the removal of BPA experienced increase and subsequently decrease with the PACl-Al(13) dosage increasing. The optimal PACl-Al(13) dosage was found at BPA/PACl-Al(13)=1:2.6(M/M) under our experiment conditions. Results show that the maximum BPA removal efficiency occurred at pH 9.0 due to the adsorption by Al(13) aggregates onto BPA rather than charge neutralization mechanism by polynuclear aluminous salts in the solution. The humic acid matters and kaolin in the solution have significant effect on BPA removal with PACl-Al(13) in the coagulation. The BPA removal will be weakened at high humic matters. The removal rate of BPA increased and subsequently decreased with the turbidity increasing. PMID- 19555960 TI - Sulfonation of ordered mesoporous carbon supported Pd catalysts for formic acid electrooxidation. AB - A novel supporting material containing benzenesulfonic acid (BSA) groups and ordered mesoporous carbons (OMCs) was first prepared by in situ radical polymerization of 4-styrenesulfonate and isoamyl nitrite under ambient conditions. Then, Pd nanoparticles were deposited on as-produced OMCs (f-OMCs) by the NaBH(4) reduction method. The structure and nature of the resulting composites were characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and nitrogen adsorption-desorption. The results show that BSA groups are created and the texture and surface chemistry are altered, whereas the ordered porous structure is maintained. The electrocatalytic properties of the Pd/f-OMCs catalysts for formic acid oxidation (HCOOH) have been investigated by cyclic voltammetry and chronoamperometry methods, and excellent electrocatalytic activity can be observed. PMID- 19555961 TI - Biomimetic synthesis of copolymer-silica nanoparticles with tunable compositions and surface property. AB - We report using cationic poly(acrylamide-co-2-(dimethylamino) ethyl methacrylate, methyl chloride quaternized) (poly(AM-co-DMC)) to mediate biomimetic synthesis of hybrid copolymer-silica nanoparticles under ambient conditions. Poly(AM-co-DMC)s with various mole contents of DMC were prepared by solution copolymerization in water. Silicification was achieved by simply stirring a mixture of tetramethyl orthosilicate and an aqueous poly(AM-co-DMC) solution at room temperature for 30 min. Copolymers-silica hybrid nanoparticles were characterized with transmission electron microscopy (TEM), FT-IR spectroscopy, (1)H NMR, thermogravimetry and aqueous electrophoresis. TEM studies indicated that the hybrid nanoparticles have well-defined spherical morphology and relatively narrow polydispersity with diameters of less than 50 nm. The compositions and zeta potentials of hybrid nanoparticles could be controlled by simply adjusting compositions of copolymers and solution conditions for silica mineralization. Due to the tunable compositions and surface zeta potentials, these new particles would be expected to have potential applications for controlled delivery, therapeutics and bioimaging. PMID- 19555962 TI - Phthalate levels and low birth weight: a nested case-control study of Chinese newborns. AB - OBJECTIVE: To assess maternal-fetal exposure to phthalates and investigate whether in utero phthalate exposure is associated with low birth weight (LBW). STUDY DESIGN: A total of 201 newborn-mother pairs (88 LBW cases and 113 controls) residing in Shanghai were enrolled in this nested case-control study during 2005 2006. Maternal blood, cord blood, and meconium specimens were collected and analyzed for phthalates by high-performance liquid chromatography-mass spectrometry. Nonparametric tests were used to compare demographic characteristics in cases and controls. Conditional logistic regression and Spearman correlation were used to analyze the association between phthalate exposure and LBW. RESULTS: No significant differences in gestational age, prepregnancy body mass index, prenatal care, vitamin supplementation, or socioeconomic levels were found between the LBW and control infants. More than 70% of the biosamples had quantifiable levels of phthalates, with higher levels in the LBW infants compared with the controls. Prenatal di-n-butyl phthalate (DBP) exposure was associated with LBW, and di-2-ethylhexyl phthalate (DEHP) was negatively associated with birth length. After adjusting for the potential confounders, DBP concentrations in the highest quartile were associated with an increased risk of LBW. CONCLUSIONS: Newborns in China are ubiquitously exposed to phthalates; significantly higher phthalate levels were detected in LBW cases compared with controls. In utero DBP and DEHP exposures were associated with LBW in a dose-dependent manner. Prenatal phthalate exposure may be a risk factor for LBW. PMID- 19555963 TI - Health-related quality of life in 5-year-old very low birth weight infants. AB - OBJECTIVE: To investigate the effect of preterm birth, the time of birth, and birth hospital level and district on health-related quality of life (HRQoL) and quality-adjusted life years (QALYs). STUDY DESIGN: This national study included all very low birth weight infants (VLBWIs; birth weight 2-fold increase following wounding at both 15 and 45 min, yet absent in the E18 groups. Furthermore, levels of cleaved PARP were also increased by >2-fold at both 15 and 45 min in E15 wound groups, whereas a relatively small amount was only seen in the E18 wound groups at 45 min. DNA-histone fragmentation ELISA assay showed a 5-fold increase in the enrichment of histone-associated DNA fragments in the E15 wounded tissue compared with the time-matched controls at 45 min. This was not seen with the E18 tissue. CONCLUSIONS: Previously, we demonstrated that cutaneous wounds in E15 fetal mice heal in a scarless manner, while similar wounds in E18 mice heal with scar formation. Results from our current work demonstrate differences in apoptosis in mid- compared with late-gestational mouse skin as well as shortly after wounding. Our results suggest that in mid-gestational wounds, activation of apoptotic pathways may be mediated through effector caspase 7 signals with inactivation of PARP. This initiation of apoptotic signals following tissue injury may play a role in scarless wound repair. PMID- 19555973 TI - Surgical education in the internet era. AB - Technological advancements, along with economic and political issues, have resulted in major changes in surgical education. The development of high fidelity simulators and the widespread availability of the Internet have allowed learning to be shifted away from the operating room. Furthermore, the Internet provides an opportunity for surgical educators to standardize general surgery training and assessment and to develop collaborations nationally and globally. This paper highlights presentations about the challenges as well as the rewards of surgical education in the age of the Internet from the 2009 Academic Surgical Congress. PMID- 19555974 TI - The impact of resident work-hour restrictions on outcomes of cardiac operations. AB - BACKGROUND: The Accreditation Council of Graduate Medical Education mandated an 80-h/wk resident physician work-hour restriction on July 1, 2003. The objective of this study was to evaluate the impact of the resident work-hour restriction on outcomes of cardiac operations. MATERIALS AND METHOD: We reviewed records of 1562 patients who underwent cardiac operations at our institution between 1997 and 2007, and we compared outcomes of operations performed before July 1, 2003 (pre reform, n=777) and those performed after July 1, 2003 (post-reform, n=785). Multivariate analysis with logistic regression was used to test for the independent effects of the resident work-hour reform by controlling for patient specific confounding factors. RESULTS: Post-reform patients had a significantly lower 30-d mortality rate (1.8% versus 3.9%; P=0.01) and a slightly lower 6-month mortality rate (4.5% versus 6.3%; P=0.12) than pre-reform patients. Multivariate analysis revealed that the post-reform patients had significantly lower adjusted 30-d mortality (odds ratio, 0.37; 95% CI, 0.18-0.75; P=0.006) and 6-mo mortality (odds ratio, 0.56; 95% CI, 0.34-0.91; P=0.02) than the pre-reform patients. CONCLUSIONS: Cardiac operations performed after the resident work-hour restriction went into effect were associated with significantly lower adjusted 30 d and 6-mo mortality rates than were operations performed before the work-hour restriction became effective. PMID- 19555975 TI - Oxidative stress implication after prolonged storage donor heart with blood versus crystalloid cardioplegia and reperfusion versus static storage. AB - Several factors are known to limit cardiac transplantation, such as number of donors, quality of cardiac graft preservation, and ischemia-reperfusion injury. Some mechanisms of reperfusion injury are now recognized; they include oxygen free radical (OFR), white blood cells activation, changes in calcium influx, alteration of microvascular blood flow, and sympathetic activation. The goal of this study was to assess the effects of two types of cardioplegia with long-term storage, either static or continuous perfusion, in 30 isolated sheep hearts as a model for heart transplantation. We examined myocardial function, histology, ischemic damage, and markers of oxidative stress. Two types of cardioplegia and storage conditions using a Langendorff reperfusion were studied in a combined approach: crystalloid (CP) [groups I and III] or cold oxygenated autologous blood (BC) [groups II and IV], immediate storage during 8h in profound hypothermia (groups I and II), or reperfused with crystalloid (group III), or blood cardioplegia (group IV). All perfusate samples were drawn from the coronary sinus. Lactate levels increased progressively in groups I, II, and IV, but not in group III, as no significant elevation was shown [90 min: 13.6+/-1.7 versus 5.2+/ 1.0 mmol/L (P<0.01)]. Arrhythmias were more frequent when using BC (n=5) than CP (n=0). For plasma thiobarbituric acid-reactive substances (TBARS) levels a significant difference was found between group III and the other groups since 15 to 90 min (P<0.05). Vitamin E concentration decreased significantly from 5 min for groups II and IV, 15 min for group I, and 30 min for group III, with a significant difference between groups II and IV (P<0.05) but not between groups I and III. CP followed by a reperfusion with the same solution showed a significantly lower ischemic injury and OFR production, less frequent ventricular arrhythmias while stable hemodynamic parameters carried on. However, this protocol did not act on the early postoperative contractile function. PMID- 19555976 TI - Cytoprotective effects of human interleukin-10 gene transfer against necrosis and apoptosis induced by hepatic cold ischemia/reperfusion injury. AB - BACKGROUND: Apoptosis as well as necrosis may play an important role in hepatic ischemia/reperfusion (I/R) injury. Interleukin 10 (IL-10), a Th2 type cytokine, modulates inflammatory responses by inhibiting the production of proinflammatory cytokines. The study focused on cytoprotective and antiapoptotic pathways to assess mechanisms by which gene transduction of human IL-10 (hIL-10) may renders grafts resistant to the cold I/R injury. MATERIALS AND METHODS: Adenoviruses encoding hIL-10 or beta-galactosidase (LacZ) were injected via the superior mesenteric vein into prospective donor animals. The donor liver was harvested 48h after transduction, and stored for 12h at 4 degrees C lactated Ringer's solution prior to being transplanted. Graft survival, liver function, the degree of necrosis and apoptosis, and the molecules of apoptotic networks were assessed. RESULTS: Ad-hIL-10 pretreatment significantly prolonged the survival of liver grafts by improving liver function, preserving hepatocyte integrity and architecture, and depressing intrahepatic apoptosis and necrosis. In addition, Ad hIL-10 pretreatment diminished the release of cytochrome c from mitochondria into cytoplasm and caspase-3 activity, with simultaneous up-regulated of antioxidant HO-1 and anti-antiapoptotic Bcl-2 molecules. CONCLUSION: Adenoviral gene transfer of hIL-10 ameliorated cold I/R injury by decreasing hepatic necrosis and apoptosis. The underlying mechanism of cytoprotective effects may at least be involved with the inhibition of caspase-3 activity and mitochondrial cytochrome c release, and the up-regulation of antiapoptotic (Bcl-2) and antioxidant (HO-1) molecules. PMID- 19555977 TI - The effects of increased extracellular deformation, pressure, and integrin phosphorylation on fibroblast migration. AB - Wound healing requires fibroblast migration. Increased pressure slows migration and ulcer healing. Pressure also induces beta1 integrin phosphorylation. We hypothesized that beta1 phosphorylation influences cell adhesion and migration. We compared the effects of increased pressure on the adhesion and motility of GD25 beta1-integrin null fibroblasts transfected with wild-type beta1A-integrin, S785A or TT788/9AA (phosphorylation-deficient), or T788D (constitutively phosphomimetic) mutants. GD25 beta1 null cells adhered less than wild type beta1A cells, suggesting adherence by non-integrin mechanisms. Preventing Ser-785 or Thr 788/789 phosphorylation reduced adhesion, suggesting that phosphorylation regulates adhesiveness. Substituting Asp for Thr788 stimulated adhesion on both substrates. Pressure decreased migration in all lines and on all matrixes, the most in wild type beta1A integrin cells and only slightly in beta1A TT788/9AA cells. In comparison, another physical force, repetitive deformation, increased migration in the beta1A integrin T788D, S785A, and wild type cells on fibronectin, and decreased migration on collagen. Deformation did not affect the migration of GD25 beta1-integrin null or TT788/9AA cells. Extracellular signal regulated kinase (ERK) blockade neither altered basal migration nor prevented pressure inhibition, while the cellular deformation response on fibronectin was altered. beta1-Integrin phosphorylation regulates cellular adhesion and the deformation effects on motility. The pressure-induced motility response is independently regulated. PMID- 19555978 TI - Interleukin-6 expression on the biliary epithelia during inflammation-associated biliary carcinogenesis in bilioenterostomized hamsters. AB - BACKGROUND: Chronic inflammatory conditions of the biliary tree strongly predispose patients to biliary carcinoma. The aim of this study was to evaluate the role of interleukin-6 (IL-6) expression during biliary carcinogenesis in bilioenterostomized hamsters. MATERIALS AND METHODS: Syrian hamsters were subjected to either a choledochoduodenostomy (CD, n=11) or a simple laparotomy (SL, n=10) and then received N-nitrosobis(2-oxopropyl)amine (BOP) treatment. The animals were sacrificed 20 wk after surgery and the development of biliary carcinoma, the presence and degree of cholangitis, and IL-6 expression on the biliary epithelia were examined histologically. RESULTS: In the CD group, eight hamsters (73%) demonstrated persistent cholangitis and six (55%) of them developed intrahepatic biliary carcinoma, while no hamster without cholangitis showed any biliary carcinoma. In the SL group, cholangitis was recognized in four hamsters (40%) and no development of biliary carcinoma was identified. A significantly high incidence of tumor development (P=0.024) and a close correlation between the presence of cholangitis and the occurrence of biliary carcinoma (P =0.013) were thus evident in the CD group. Moreover, the degree of cholangitis was significantly higher in the CD hamsters (P=0.041) and an IL-6 overexpression was identified in five hamsters that had undergone a CD, with a scattered expression on the intra- and extrahepatic biliary epithelia. Despite the fact that the induced biliary carcinomas showed a multicentric occurrence in the liver, these tumors originated from within the restricted area where IL-6 was expressed. CONCLUSIONS: A deregulated IL-6 overexpression on the biliary epithelia may therefore be involved in inflammation-associated biliary carcinogenesis in hamsters that have undergone a bilioenterostomy. PMID- 19555980 TI - A comparison of autologous and allogenic bone marrow-derived mesenchymal stem cell transplantation in canine spinal cord injury. AB - The purpose of this study is to compare the therapeutic effects between autologous and allogenic bone-marrow-derived mesenchymal stem cell (MSC) transplantation in experimentally-induced spinal cord injury (SCI) of dogs. Thirty adult Beagle dogs (control group=10, autologous group=10, and allogenic group=10) were used in this study. Prelabeled MSCs were intrathecally transplanted through the lumbar spinal cord into the injured lesion at a density of 1 x 10(7) cells 7 days after SCI. Neurological signs of dogs in both autologous and allogenic groups were improved in their pelvic limbs after SCI compared with those in control group. Both autologous and allogenic groups showed significantly higher the Olby scores than control group (p<0.05). This finding was consistent with results of MRI and histopathological examination in both groups. Immunofluorescence analysis revealed that prelabeled autologous and allogenic MSCs were detected in the injured lesions both at 1 and 4 weeks after transplantation. However, the distribution ratio of MSCs on the injured lesion in allogenic group was significantly decreased at 4 weeks after transplantation relatively to at 1 week after transplantation. The mRNA expression for neurotrophic factors in both allogenic and autologous groups was significantly higher than that in control groups (p<0.05). Even though autologous MSC transplantation showed more beneficial effect than that of allogenic MSC transplantation, transplantation of allogenic MSCs also improved functional recovery following SCI. This study demonstrates that both autologous and allogenic MSC transplantation could be clinically useful therapeutic approaches for treating SCI. PMID- 19555981 TI - [Pulmonary embolism during pregnancy: CT or lung scan?]. PMID- 19555982 TI - [Acute ischemia in the basilar artery territory]. PMID- 19555983 TI - Reactive electrophilic oxylipins: pattern recognition and signalling. AB - Oxidized lipids in plants comprise a variety of reactive electrophiles that contain an alpha,beta-unsaturated carbonyl group. While some of these compounds are formed enzymatically, many of them are formed by non-enzymatic pathways. In addition to their chemical reactivity/toxicity low levels of these compounds are also biologically active. Despite their structural diversity and biosynthetic origin, common biological activities such as induction of defense genes, activation of detoxification responses and growth inhibition have been documented. However, reactive electrophilic oxylipins are poorly defined as a class of compounds but have at least two properties in common, i.e., lipophilicity and thiol-reactivity. Thiol-reactivity is a property of reactive oxylipins (RES) shared by reactive oxygen and nitrogen species (ROS and RNS) and enables these agents to modify proteins in vivo. Thiol-modification is assumed to represent a key mechanism involved in signal transduction. A metaanalysis of proteomic studies reveals that RES oxylipins, ROS and RNS apparently chemically modify a similar set of highly sensitive proteins, virtually all of which are targets for thioredoxins. Moreover, most of these proteins are redox-regulated, i.e., posttranslational thiol-modification alters the activity or function of these proteins. On the transcriptome level, effects of RES oxylipins and ROS on gene induction substantially overlap but are clearly different. Besides electrophilicity other structural properties such as target affinity apparently determine target selectivity and biological activity. In this context, different signalling mechanisms and signal transduction components identified in plants and non-plant organisms as well as putative functions of RES oxylipins are discussed. PMID- 19555985 TI - Relationship between coffee cultivation practices in Colombia and exposure to infection with Leishmania. AB - The inhabitants of coffee-growing municipalities consistently report the highest annual rates of cutaneous leishmaniasis in Colombia. During the last two decades most Colombian coffee growers have changed from the traditional system of cultivation, where the crop is grown under different species of shade trees, to an intensified system where it is grown at high densities in full sunlight. This change may affect transmission of Leishmania spp. to humans in several ways, probably resulting from reduced human-vector contact. The responses of residents of traditional and intensified coffee plantations to the leishmanin skin test were compared to ascertain whether intensification has indeed affected Leishmania transmission. Although prevalence of infection was significantly higher (P< or =0.01) among residents of traditional plantations (26.8%) than among those of intensified ones (13.2%), no significant difference could be demonstrated with respect to incidence of infection at the time of the study. Similar rates of infection were found for men and women, although the incidence of infection was significantly higher among the latter in intensified plantations. Changes to the type of data collected and the data collection process will facilitate the evaluation of the long-term effects of intensification of coffee plantations on Leishmania transmission. PMID- 19555984 TI - [Gingivoperiosteoplasty associated to bone graft: radiological evaluation]. AB - INTRODUCTION: Gingivoperiosteoplasty associated to bone graft is part of a therapeutic strategy applied to the first 20 years of a patient's life. Management is pluridisciplinary. Most authors recommend a bone graft in mixed dentition at the end of premaxillary growth. Retroalveolar and panoramic radiography are the most often used to assess the bone height of the grafted site. We retrospectively studied the radiographies of 57 alveolar grafts in 44 patients. MATERIAL AND METHODS: Between 1999 and 2005, 44 patients underwent gingivoperiosteoplasty associated to bone graft. Thirteen underwent bilateral reconstruction. The surgical interventions were performed by the same surgeon. One year after surgery, the panoramic radiographies were analyzed by a single expert. The bone height compared to roots of adjacent teeth was classified in four grades. Grades 1 and 2 were considered as satisfactory or good and grades 3 and 4 not satisfactory and an indication for a new bone graft. In case of bilateral cleft, each side was analyzed independently. RESULTS: Grades 1 and 2 accounted for 84.2% of grafts. There was no statistical difference in alveolar bone height between patients presenting with agenesis of the lateral incisive. Eighty-one percent of patients grafted with mixed dentition (66% of the cases) had satisfactory results (35% of grade 1 and 46% of grade 2). Patients operated on after 15 years (n=15) had good results in 75% of the cases, 33% were bilateral cleft patients. There was no statistical difference between patients operated on early and those with delayed surgery. DISCUSSION: Radiological results for gingivoperiosteoplasty associated to bone graft are satisfactory. The procedure is easy, cheap, and reproducible. Evaluation with panoramic radiography is not as accurate as with the Denta Scan. CT scan is not used systematically to follow up alveolar cleft palate in children so as to limit irradiation. Volumetric tomography (cone beam) may be the best assessment. PMID- 19555987 TI - Mathematical modelling of hydrogen sulphide emission and removal in aerobic biofilters comprising chemical oxidation. AB - Four different empirical expressions have been compared for estimating the removal of hydrogen sulphide (H(2)S) from wastewater by chemical oxidation during its treatment in an aerated biofilter. The relative importance of this removal process is considered in a mass balance proposed by an emission model. Two of the four models investigated were able to predict the mean H(2)S removed fraction within a confidence interval of 95% and they demonstrated good agreement with experimental data. Biodegradation and oxidation were the two main removal mechanisms in the biofilter whereas stripping and volatilization made only minor contributions. However they can be of significance when the emission rates are calculated. PMID- 19555986 TI - Nef-mediated MHC class I down-regulation unmasks clonal differences in virus suppression by SIV-specific CD8(+) T cells independent of IFN-gamma and CD107a responses. AB - CD8(+) T lymphocytes (CTL) play a role in controlling HIV/SIV infection. CTL antiviral activity is dependent on recognition of antigenic peptides associated with MHC class I molecules on infected target cells, and CTL activation can be impaired by Nef-mediated down-regulation of MHC class I molecules. We tested the ability of a series of rhesus macaque CD8(+) T-cell clones specific for the SIV Gag CM9 peptide to suppress SIV infection of autologous CD4(+) T cells. We used a set of SIV(mac)239 viruses with either wild-type Nef or Nef mutations that impair MHC class I down-regulation. All CTL clones efficiently suppressed virus replication in cells infected with mutant viruses with altered Nef function, phenotypically MHC class I(high) or MHC class I(intermediate). However, the ability of the clones to suppress virus replication was variably reduced in the presence of wild-type Nef (MHC class I(low)) despite the observations that all CTL clones showed similar IFN-gamma responses to titrated amounts of cognate peptide as well as to SIV-infected cells. In addition, the CTL clones showed variable CD107a (CTL degranulation marker) responses that did not correlate with their capacity to suppress virus replication. Thus, the clonal differences are not attributable to TCR avidity or typical effector responses, and point to a potential as yet unknown mechanism for CTL-mediated suppression of viral replication. These data emphasize that current assays for evaluating CTL responses in infected or vaccinated individuals do not fully capture the complex requirements for effective CTL-mediated control of virus replication. PMID- 19555988 TI - Waste activated sludge hydrolysis and short-chain fatty acids accumulation under mesophilic and thermophilic conditions: effect of pH. AB - The effect of pH (4.0-11.0) on waste activated sludge (WAS) hydrolysis and short chain fatty acids (SCFAs) accumulation under mesophilic and thermophilic conditions were investigated. The WAS hydrolysis increased markedly in thermophilic fermentation compared to mesophilic fermentation at any pH investigated. The hydrolysis at alkaline pHs (8.0-11.0) was greater than that at acidic pHs, but both of the acidic and alkaline hydrolysis was higher than that pH uncontrolled under either mesophilic or thermophilic conditions. No matter in mesophilic or thermophilic fermentation, the accumulation of SCFAs at alkaline pHs was greater than at acidic or uncontrolled pHs. The optimum SCFAs accumulation was 0.298g COD/g volatile suspended solids (VSS) with mesophilic fermentation, and 0.368 with thermophilic fermentation, which was observed respectively at pH 9.0 and fermentation time 5 d and pH 8.0 and time 9 d. The maximum SCFAs productions reported in this study were much greater than that in the literature. The analysis of the SCFAs composition showed that acetic acid was the prevalent acid in the accumulated SCFAs at any pH investigated under both temperatures, followed by propionic acid and n-valeric acid. Nevertheless, during the entire mesophilic and thermophilic fermentation the activity of methanogens was inhibited severely at acid or alkaline pHs, and the highest methane concentration was obtained at pH 7.0 in most cases. The studies of carbon mass balance showed that during WAS fermentation the reduction of VSS decreased with the increase of pH, and the thermophilic VSS reduction was greater than the mesophilic one. Further investigation indicated that most of the reduced VSS was converted to soluble protein and carbohydrate and SCFAs in two fermentations systems, while little formed methane and carbon dioxide. PMID- 19555989 TI - Sludge properties and their effects on membrane fouling in submerged anaerobic membrane bioreactors (SAnMBRs). AB - Two submerged anaerobic membrane bioreactors (SAnMBRs) (thermophilic vs. mesophilic) were operated for a period of 3.5 months with kraft evaporator condensate at a feed chemical oxygen demand of 10,000 mg/L. The results show that the filtration behavior of the two systems was significantly different. The filtration resistance in the thermophilic SAnMBR was about 5-10 times higher than that of the mesophilic system when operated under similar hydrodynamic conditions. Comparison of sludge properties and cake layer structure from the two systems was made to elucidate major factors governing the different filtration characteristics. There were more soluble microbial products (SMP) and biopolymer clusters (BPC) produced and a larger portion of fine flocs (<15 microm) in the thermophilic SAnMBR. Analysis of bound extracellular polymeric substances (EPS) showed that the thermophilic sludge had a higher protein/polysaccharide ratio in EPS, as compared to that in the mesophilic sludge. A series of analyses, including Fourier transform infrared (FTIR) spectroscopy, energy dispersive X-ray spectroscopy (EDX), confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), atomic force microscopy (AFM) and particle size analyzer showed that the cake layer formed in the thermophilic SAnMBR contained higher levels of both organic and inorganic foulants, smaller particle sizes, and especially, a denser and more compact sludge cake structure. These results indicate that floc size, SMP, BPC, bound EPS as well as cake layer structure are the major factors governing membrane fouling in SAnMBR systems. PMID- 19555990 TI - Effect of COD/SO(4)2- ratio and Fe(II) under the variable hydraulic retention time (HRT) on fermentative hydrogen production. AB - The effect of chemical oxygen demand/sulfate (COD/SO(4)(2-)) ratio on fermentative hydrogen production using enriched mixed microflora has been studied. The chemostat system maintained with a substrate (glucose) concentration of 15 g COD L(-1) exhibited stable H(2) production at inlet sulfate concentrations of 0-20 g L(-1) during 282 days. The tested COD/SO(4)(2-) ratios ranged from 150 to 0.75 (with control) at pH 5.5 with hydraulic retention time (HRT) of 24, 12 and 6h. The hydrogen production at HRT 6h and pH 5.5 was not influenced by decreasing the COD/SO(4)(2-) ratio from 150 to 15 (with control) followed by noticeable increase at COD/SO(4)(2-) ratios of 5 and 3, but it was slightly decreased when the COD/SO(4)(2-) ratio further decreased to 1.5 and 0.75. These results indicate that high sulfate concentrations (up to 20,000 mg L( 1)) would not interfere with hydrogen production under the investigated experimental conditions. Maximum hydrogen production was 2.95, 4.60 and 9.40 L day(-1) with hydrogen yields of 2.0, 1.8 and 1.6 mol H(2) mol(-1) glucose at HRTs of 24, 12 and 6h, respectively. The volatile fatty acid (VFA) fraction produced during the reaction was in the order of butyrate>acetate>ethanol>propionate in all experiments. Fluorescence In Situ Hybridization (FISH) analysis indicated the presence of Clostridium spp., Clostridium butyricum, Clostridium perfringens and Ruminococcus flavefaciens as hydrogen producing bacteria (HPB) and absence of sulfate reducing bacteria (SRB) in our study. PMID- 19555991 TI - Enhancing anaerobic digestibility and phosphorus recovery of dairy manure through microwave-based thermochemical pretreatment. AB - Anaerobic digestion and struvite precipitation are two effective ways of treating dairy manure for recovering biogas and phosphorus. Anaerobic digestion of dairy manure is commonly limited by slow fiber degradation, while struvite precipitation is limited by the availability of orthophosphate. The aim of this work is to study the possibility of using microwave-based thermochemical pretreatment to simultaneously enhance manure anaerobic digestibility (through fiber degradation) and struvite precipitation (through phosphorus solubilization). Microwave heating combined with different chemicals (NaOH, CaO, H(2)SO(4), or HCl) enhanced solubilization of manure and degradation of glucan/xylan in dairy manure. However, sulfuric acid-based pretreatment resulted in a low anaerobic digestibility, probably due to the sulfur inhibition and Maillard side reaction. The pretreatments released 20-40% soluble phosphorus and 9-14% ammonium. However, CaO-based pretreatment resulted in lower orthophosphate releases and struvite precipitation efficiency as calcium interferes with phosphate to form calcium phosphate. Collectively, microwave heating combined with NaOH or HCl led to a high anaerobic digestibility and phosphorus recovery. Using these two chemicals, the performance of microwave- and conventional-heating in thermochemical pretreatment was further compared. The microwave heating resulted in a better performance in terms of COD solubilization, glucan/xylan reduction, phosphorus solubilization and anaerobic digestibility. Lastly, temperature and heating time used in microwave treatment were optimized. The optimal values of temperature and heating time were 147 degrees C and 25.3 min for methane production, and 135 degrees C and 26 min for orthophosphate release, respectively. PMID- 19555993 TI - Mercury degassing from forested and open field soils in Rondonia, Western Amazon, Brazil. AB - A Teflon dynamic flux chamber was used to characterize Gaseous Elemental Mercury (GEM) flux from forested and open field soils in a highly changing environment in Rondonia State, western Amazon. We simultaneously analyzed meteorological parameters at the soil level relating GEM fluxes to soil temperature, air humidity, soil moisture, solar radiation, and speed and wind direction. We also examined variations of atmospheric GEM concentration. GEM fluxes during the day and night in the open field site were significantly different (17+/-14ngm(-2) h( 1) and 0.9+/-1.9ngm(-2)h(-1), for day and night, respectively), but were similar within the forest site (4.8+/-1.4ngm(-2)h(-1) and 4.4+/-1.8ngm(-2) h(-1) for day and night periods, respectively). A comparison between 24-h periods averages in the two sites showed much larger emission from the open field site. GEM fluxes at the open field site were positively correlated with soil moisture, solar irradiation and soil temperature and inversely correlated with air humidity. At the forest site GEM fluxes showed no correlation with meteorological variables. At the open field site GEM concentrations significantly correlated with GEM flux, at least during the day. At night in the open field site and during the day and night at the forest site no correlation was found between GEM fluxes and GEM concentrations in the ambient air. Higher emissions from the open field site support earlier studies showing larger Hg remobilization following forest conversion to pasture. PMID- 19555992 TI - Application of real-time PCR assays to genotyping of F-specific phages in river water and sediments in Japan. AB - Genotyping of F-specific RNA phages is currently one of the most promising approaches to differentiate between human and animal fecal contamination in aquatic environments. In this study, a total of 18 river water and sediment samples were collected from the Tonegawa River basin, Japan, in order to describe the genogroup distribution of F-specific RNA and DNA phages using genogroup specific real-time PCR assays. F-specific phages were detected in nine (100%) river water and six (67%) sediment samples. Eighty-five phage plaques were isolated from these samples and subjected to real-time PCR assays specific for the phages. F-specific RNA phages of human genogroups (II and III) were detected in 32 (38%) plaques, whereas those of animal genogroups (I and IV) were detected in 17 (20%) plaques. No correlation was observed between the genogroup distribution of F-specific RNA phages and the occurrence of human adenovirus genomes, suggesting that genotyping of the phages alone is inadequate for the evaluation of the occurrence of viruses in aquatic environments. SYBR Green-based real-time PCR assay revealed the presence of F-specific DNA phages in four (5%) plaques, which were further classified into two genogroups (fd- and f1-like phages) by sequence analysis. Thirty-two (38%) plaques were not classified as the F-specific phage genogroups, indicating the limited applicability of these real time PCR assays to a wide range of aquatic environmental samples worldwide. PMID- 19555994 TI - Chronic exposure to sublethal hexavalent chromium affects organ histopathology and serum cortisol profile of a teleost, Channa punctatus (Bloch). AB - Effects of chronic exposures (one and two months) to sublethal doses of hexavalent chromium (2 and 4 mg/L potassium dichromate) on organ histopathology and serum cortisol profile were investigated and their overall impact on growth and behavior of a teleost fish, Channa punctatus was elucidated. Histopathological lesions were distinct in the vital organs gill, kidney and liver. The gill lamellae became lifted, fused, and showed oedema. Hyperplasia and hypertrophy of lamellar epithelial cells were distinct with desquamation. Hypertrophy of epithelial cells of renal tubules and reduction in tubular lumens were observed in the trunk kidney. The atrophy of the head kidney interrenal cells and decreased serum cortisol level indicated exhaustion of interrenal activity. Hepatocyte vacuolization and shrinkage, nuclear pyknosis and increase of sinusoidal spaces were observed in the liver. Abnormal behavioral patterns and reduced growth rate were also noticed in the exposed fish. The chronic hexavalent chromium exposure thus by affecting histopathology of gill, kidney (including interrenal tissue) and liver could impair the vital functions of respiration, excretion, metabolic regulation and maintenance of stress homeostasis which in the long-run may pose serious threat to fish health and affect their population. PMID- 19555995 TI - Ovarian hormones control the changing expression of claudins and occludin in rat uterine epithelial cells during early pregnancy. AB - Regulation of the uterine luminal environment is important for successful attachment and implantation of the blastocyst. The contents and volume of luminal fluid are regulated in part by the tight junctions. Using immunofluorescence microscopy, protein and RNA analysis, the cellular distributions of tight junction components claudins and occludin were observed during early pregnancy and under various hormonal regimens. Results indicate that occludin and claudin-4 distribution changed during early pregnancy and in response to ovarian hormones. At the time of implantation and in response to progesterone administration to ovariectomised rats, occludin and claudin-4 showed increased immunolabelling in luminal epithelium. Interestingly, occludin protein detection in uterine luminal epithelial cells at the time of implantation was statistically significantly decreased at the time of implantation compared to day 1 of pregnancy. This suggests that a cytoplasmic pool of occludin is present at day 1 of pregnancy and is redistributed to the tight junctions at the time of implantation. The presence of occludin and claudin-4 in the tight junctions at the time of implantation and in response to progesterone suggests that the paracellular pathway is impermeable to water and Na(+) at this time, and that the transport of such substances takes place via the transcellular pathway. PMID- 19555997 TI - Extended left colon resections as part of complete cytoreduction for ovarian cancer: tips and considerations. AB - OBJECTIVES: Colectomy is a common part of cytoreductive surgery. The most common segment of the colon resected is the rectosigmoid. Extended left-sided colectomies are rarely performed. With increased emphasis on the importance of maximal cytoreductive surgery it is becoming more important to understand the outcomes related to such a procedure. The purpose of this report is to evaluate the quality of life issues related to extended left colectomies. METHODS: Data on all patients who underwent cytoreductive surgery between April 2007 and April 2009 was prospectively recorded. Nineteen patients underwent extended left colon resections. The data from these cases was evaluated with particular attention directed at the quality of life issues surrounding postoperative bowel function, and tolerance of postoperative chemotherapy. RESULTS: Nine underwent resection of the left hemicolon plus the hepatic flexure. Six underwent resection of the left hemicolon, hepatic flexure, and the ascending colon. Four underwent subtotal colectomies. Temporary diverting loop ileostomies were performed on 18 of the 19 patients each of whom consented for intraperitoneal chemotherapy. No delays in chemotherapy were observed. Median number of bowel movements at 6, 9 and 12 months were 2, 2 and 1, respectively. No fecal incontinence was observed. Patients expressed satisfaction with their bowel surgery and denied any significant decrease in their quality of life due to their bowel function. CONCLUSION: Extended left colon resection is a reasonable technique to include in patients requiring maximal cytoreduction. Quality of life related to bowel function is acceptable and chemotherapy is not delayed. PMID- 19555996 TI - Total laparoscopic radical hysterectomy for locally advanced cervical carcinoma (stages IIB, IIA and bulky stages IB) after concurrent chemoradiation therapy: surgical morbidity and oncological results. AB - OBJECTIVES: To evaluate the surgical outcome and the oncologic results of total laparoscopic radical hysterectomy (TLRH) after neoadjuvant chemoradiation therapy (CRT) for locally advanced cervical carcinoma. METHODS: All patients who underwent TLRH after CRT for stages IIB-IIA and bulky IB diseases were reviewed. The control group for this analysis was a cohort of patients treated with abdominal radical hysterectomy (ARH) after CRT for the same stage cancers. RESULTS: We reviewed 102 patients operated on between 2000 and 2008 (46 TLRH and 56 ARH). Mean age at diagnosis was 44 years, and mean B.M.I was 22.1. There was no difference in tumor characteristics between the two groups. Seven patients in the laparoscopic group required conversion to laparotomy (15%). Mean estimated blood loss (200 vs. 400 mL, p<0.01) and the median duration of hospital stay (5 vs. 8 days, p<0.01) were significantly lower in the laparoscopic group. Morbidity rates and urinary complications were reduced in the laparoscopic group (p=0.04). Local recurrence rates, disease-free and overall survival were comparable in the two groups. Best survival was observed for patients with pathological complete response or microscopic residual disease compared to patients with macroscopic residues (p<0.01). CONCLUSIONS: Radical hysterectomy after CRT is known to be difficult with significant morbidity rates and remains controversial in comparison to exclusive CRT. TLRH after preoperative CRT is feasible for patients with locally advanced cervical cancer in 85% of the cases. For these patients, TLRH compared with ARH was associated with favorable surgical outcome with comparable oncological results. PMID- 19555998 TI - Elevated aromatase expression correlates with cervical carcinoma progression. AB - OBJECTIVES: We have previously demonstrated that aromatase mRNA is induced in cervical carcinomas compared to normal tissue, suggesting that in situ aromatase expression leading to elevated local estrogen production may contribute to cervical carcinogensis. Our objectives are to examine 1) whether aromatase protein and activity are induced in cervical carcinomas, 2) aromatase expression correlates with disease stage, and 3) inflammatory cytokines (e.g., IL-6 and TNFalpha) may correlate with aromatase expression. METHODS: RNA and protein were isolated from human cervical carcinomas and normal cervical biopsies to examine aromatase expression, using real-time RT-PCR, Western blot analysis, and immunohistochemistry. Aromatase activity in tissue was measured using the tritiated water release method. IL-6 and TNFalpha expression was also examined. RESULTS: Aromatase protein and activity levels were increased in cervical carcinomas compared to normal tissue. RNA levels correlated significantly with disease progression, with highest aromatase expression detected in stage IV tumors (p<0.001, R(2)=0.77). Aromatase promoters 1.3 and 1.4 were elevated in cervical carcinomas and in cervical cancer cells. The expression of inflammatory cytokines IL-6 and TNFalpha, known to induce aromatase, significantly correlated with aromatase expression (R(2)>0.9). TNFalpha treatment induced aromatase expression in cervical cancer cells. CONCLUSION: Increased aromatase protein and activity in cervical carcinomas and the correlation of its expression with disease stage implicates it in cervical carcinogenesis. The correlation of IL-6 and TNFalpha expression with aromatase suggests that these inflammatory cytokines may induce aromatase expression, which is confirmed by induction of aromatase expression due to TNFalpha treatment of cervical cancer cells. PMID- 19556000 TI - Effects of N-vinylpyrrolidone (NVP) containing polyelectrolytes on surface properties of conventional glass-ionomer cements (GIC). AB - It has been found that polyacids containing an N-vinylpyrrolidinone (NVP) comonomer produces a glass inomer cement with improved mechanical and handling properties. The objective of this study was to investigate the effect of NVP modified polyelectrolytes on the surface properties and shear bond strength to dentin of glass ionomer cements. Poly(acrylic acid (AA)-co-itaconic acid (IA)-co N-vinylpyrrolidone) was synthesized by free radical polymerization. The terpolymer was characterized using (1)H NMR, FTIR spectroscopy and viscometry for solution properties. The synthesized polymers were used in glass ionomer cement formulations (Fuji II commercial GIC). Surface properties (wettability) of modified cements were studied by water contact angle measurements as a function of time. Work of adhesion values of different surfaces was also determined. The effect of NVP modified polyacid, on bond strength of glass-ionomer cement to dentin was also investigated. The mean data obtained from contact angle and bonding strength measurements were subjected to one- and two-way analysis of variance (ANOVA) at alpha=0.05. Results showed that NVP modified glass ionomer cements showed significantly lower contact angles (theta=47 degrees) and higher work of adhesion (WA=59.4 erg/cm(2)) in comparison to commercially available Fuji II GIC (theta=60 degrees and WA=50.3 erg/cm(2), respectively). The wettability of dentin surfaces conditioned with NVP containing terpolymer was higher (theta=21 degrees, WA=74.2 erg/cm(2)) than dentin conditioned with Fuji conditioner (theta=30 degrees, WA=69 erg/cm(2)). The experimental cement also showed higher but not statistically significant values for shear bond strength to dentin (7.8 MPa), when compared to control group (7.3 MPa). It was concluded that NVP containing polyelectrolytes are better dentin conditioners than the commercially available dentin conditioner (Fuji Cavity Conditioner, GC). NVP containing terpolymers can enhance the surface properties of GICs and also increase their bond strength to the dentin. PMID- 19556001 TI - Participation in civil society and political life among young people in Maharashtra: Findings from the Youth in India - Situation and Needs study. AB - Youth participation in civil society and political life is increasingly recognised to be an important development objective. Nonetheless, research that sheds light on the extent to which youth participate in these arenas, and the factors that facilitate or inhibit such participation remain limited in most developing countries including India. Drawing on data from a representative survey of young people in the state of Maharashtra, India, this paper explores the extent of and the factors associated with youth participation in civil society, their adherence to pro-social values, and their participation in political processes. Findings suggest that for many youth, particularly for young women, opportunities to engage in civil society and political life are limited, and that pro-social values are not uniformly observed. Findings underline the importance of education, agency and close parental interaction in facilitating youth participation in civil society and political life and their expression of pro-social values. PMID- 19556002 TI - Stressful life events and the tripartite model: relations to anxiety and depression in adolescent females. AB - Although the tripartite model reliably distinguishes anxiety and depression in adolescents, it remains unclear how negative affectivity (NA) and positive affectivity (PA) influence developmental pathways to internalizing problems. Based on models which propose that affectivity shapes how youth react to stress, the present study attempted to investigate the relative roles of NA, PA, and stressful life events in characterizing and differentiating adolescent anxiety and depression. A sample of adolescent females (N=63), including a sub-sample of adolescent mothers, completed measures of NA, PA, negative life event (NLE) occurrence, anxiety, and depression. Findings supported the tripartite model as a "temperamental reactivity to stress" approach. Anxious and depressive symptoms were predicted by a combination of high NA and high NLE occurrence. However, a combination of low PA and high NLE occurrence was uniquely linked to greater depressive symptoms. Implications of these findings for early identification and prevention programs are discussed. PMID- 19555999 TI - Serological response to an HPV16 E7 based therapeutic vaccine in women with high grade cervical dysplasia. AB - PURPOSE: Infection with oncogenic human papillomaviruses has been linked to the development of cervical neoplasia and cancer. The exclusive expression of E7, a viral oncogene, in infected cells makes this protein an ideal target for immunotherapy. We recently reported on the results of a trial in women with cervical carcinoma-in-situ using HspE7, a protein vaccine consisting of full length HPV16 E7 linked to a heat shock protein from M. bovis. The stimulating effects of HspE7 on specific cytotoxic T lymphocytes have been demonstrated in vitro and in (pre-)clinical trials. The induction of a B-cell response by HspE7 and its association with clinical outcome is unknown, and is the purpose of this study. EXPERIMENTAL DESIGN: We measured the serum IgG levels against HPV16 E7 and HPV16 and -18 VLPs using a multiplexed Luminex based assay in 57 women with CIS who received the HspE7 vaccine. RESULTS: Vaccination with HspE7 results in a modest, yet maintained increase in HPV16 E7 specific IgG levels. While not significant, increased HPV16 E7 IgG levels appear to be correlated with a positive therapeutic effect. Women who were previously treated for recurrent disease (by LEEP) had significantly higher HPV16 E7 IgG levels compared with subjects without recurrent disease (p=0.01). In women with recurrent disease, higher IgG levels correlated with complete pathological response. CONCLUSIONS: This study suggests that IgG levels could potentially be used as a marker for response to a therapeutic vaccine. Further translational investigations of the 'priming' of local immune responses using extirpative procedures should be explored. PMID- 19556003 TI - The enhancement of dermal papilla cell aggregation by extracellular matrix proteins through effects on cell-substratum adhesivity and cell motility. AB - Generally, cells tend to aggregate on a substratum with lower cell adhesivity. However, it also leads to compromised cell growth and higher cell loss after seeding. This study is aimed at tackling this dilemma by extracellular matrix (ECM) protein coating of a lower adhesive substratum poly(ethylene-co-vinyl alcohol) (EVAL) that has been shown to facilitate hair follicle dermal papilla (DP) spheroid formation. We found that coating with either fibronectin (Fn), collagen I, or collagen IV yields higher adhesivity and cell growth than that with laminin. However, cells can only aggregate on uncoated or Fn-coated EVAL. Quantitatively, Fn coating increases the number of spheroids by 67%. Analysis of cell migration reveals that collagen I, collagen IV and laminin coatings reduce cell motility, while Fn coating keeps cells highly motile. Inhibition of cell migration hinders spheroid formation. In addition, disruption of Fn function does not significantly compromise intercellular adhesion. Hence, Fn enhances cell aggregation by enhancing cell attachment, cell growth and cell motility. Our study demonstrates that intercellular organization as spheroids or flat monolayers is switchable by specific ECM protein coating and preserving cell motility is vital to cell aggregation. In addition to generation of spheroidal DP microtissues for hair follicle regeneration and large-scale production of aggregates of other cells, this strategy can help to regulate the tissue substrate adhesivity and tissue spreadability on the surface of implantable materials. PMID- 19556004 TI - Involvement of neuropeptide Y in the acute, chronic and withdrawal responses of morphine in nociception in neuropathic rats: behavioral and neuroanatomical correlates. AB - Although morphine is a potent antinociceptive agent, its chronic use developed tolerance in neuropathic pain (NP). Furthermore, opioid antagonist naloxone attenuated the antinociceptive effect of neuropeptide Y (NPY). The present study investigated the role of NPY and NPY Y1/Y5 receptors in acute and chronic actions of morphine in neuropathic rats using thermal paw withdrawal test and immunocytochemistry. In acute study, intracerebroventricular (icv) administration of morphine, NPY or NPY Y1/Y5 receptors agonist [Leu(31),Pro(34)]-NPY produced antinociception, whereas selective NPY Y1 receptors antagonist BIBP3226 caused hyperalgesia. While NPY or [Leu(31),Pro(34)]-NPY potentiated, BIBP3226 attenuated morphine induced antinociception. Chronic icv infusion of morphine via osmotic minipumps developed tolerance to its antinociceptive effect, and produced hyperalgesia following withdrawal. However, co-administration of NPY or [Leu(31),Pro(34)]-NPY prevented the development of tolerance and withdrawal hyperalgesia. Sciatic nerve ligation resulted in significant increase in the NPY immunoreactive (NPY-ir) fibers in ventrolateral periaqueductal gray (VLPAG) and locus coeruleus (LC); fibers in the dorsal part of dorsal raphe nucleus (DRD) did not respond. While chronic morphine treatment significantly reduced NPY-ir fibers in VLPAG and DRD, morphine withdrawal triggered significant augmentation in NPY immunoreactivity in the VLPAG. NPY-immunoreactivity profile of LC remained unchanged in all the morphine treatment conditions. Furthermore, removal of sciatic nerve ligation reversed the effects of NP, increased pain threshold and restored NPY-ir fiber population in VLPAG. NPY, perhaps acting via Y1/Y5 receptors, might profoundly influence the processing of NP information and interact with the endogenous opioid system primarily within the framework of the VLPAG. PMID- 19556005 TI - Third-party mesenchymal stem cells as part of the management of graft-failure after haploidentical stem cell transplantation. PMID- 19556006 TI - Molecular hallmarks of anti-chromatin antibodies associated with the lupus susceptibility locus, Sle1. AB - Anti-nuclear antibodies constitute the hallmark of lupus. The NZM2410-derived Sle1 lupus susceptibility interval on murine chromosome 1 breaches tolerance, leading to the emergence of anti-nuclear autoantibodies targeting nucleosomes. However, little is known about the molecular structure of the anti-nucleosome autoantibodies from this genetically simplified mouse model of lupus. In this study, the immunoglobulin heavy chain and light chain sequences of 50 anti nuclear monoclonal antibodies derived from five B6.Sle1(z) mice were compared to non-nuclear antibody controls. Compared to two different sets of non-nuclear antibodies, anti-nucleosome antibodies derived from B6.Sle1(z) congenic mice exhibited a high degree of clonal expansion and three distinct sequence motifs in their heavy chains - cationic CDR3 stretches, non-anionic CDR2 regions, and an increased frequency of aspartate residues at H50, which together increased the likelihood of an antibody being chromatin-reactive by approximately 4-fold. PMID- 19556007 TI - Role of RDBP and SKIV2L variants in the major histocompatibility complex class III region in polypoidal choroidal vasculopathy etiology. AB - PURPOSE: To investigate whether polymorphisms in 4 tightly linked genes of the major histocompatibility complex class III--complement component 2 (C2), complement factor B (CFB), RD RNA-binding protein (RDBP), and superkiller viralicidic activity 2-like (SKIV2L)--are associated with polypoidal choroidal vasculopathy (PCV). DESIGN: Cross-sectional study. PARTICIPANTS: A case-control group of 136 PCV subjects and 183 unrelated controls. METHODS: We performed an association analysis between PCV and polymorphisms across the C2-CFB-RDBP-SKIV2L region in a Japanese population, genotyping 13 single nucleotide polymorphisms (SNPs) spanning this region, including rs9332739 (E318D), rs547154, rs4151667 (L9H), and rs641153 (R32Q) that are known to be associated with age-related macular degeneration (AMD). Genotyping was conducted using TaqMan technology (Applied Biosystems, Foster City, CA). We also examined population stratification in our study cohort. MAIN OUTCOME MEASURES: Allele and haplotype frequencies of the variants across the C2-CFB-RDBP-SKIV2L region. RESULTS: We initially scanned the C2-CFB locus using 11 SNPs that capture the majority of common variations in this locus. We found a significant omnibus haplotype association and a single disease-protective haplotype, but individually, none of the 11 SNPs were associated with PCV. Further studies led to the identification of 2 untested allelic variants in RDBP (rs3880457) and SKIV2L (rs2075702) that were located on the protective haplotype. We also analyzed these 2 SNPs, detecting a significant association with a decreased risk of developing PCV (for both SNPs, allelic P = 0.0038 and per allele odds ratio = 0.31 [95% confidence interval, 0.13-0.71]). The 2 SNPs were correlated (r(2) = 1) in our dataset. Haplotype analysis and conditional testing demonstrated that either rs3880457 or rs2075702 could fully account for the omnibus haplotype association detected across the C2-CFB-RDBP SKIV2L region. Population stratification analyses excluded stratification artifacts in our study cohort. CONCLUSIONS: Our results do not support any major role of the 4 AMD-associated variants in the risk of developing PCV, but favor a predominant association with the RDBP-SKIV2L variants, which has some potential implications for pathobiological differences between PCV and neovascular AMD. Further genetic characterization of this locus will provide additional insights into the genetic basis of PCV susceptibility. PMID- 19556009 TI - Cyclothymic temperament and major depressive disorder: a study on Italian patients. AB - BACKGROUND: Classical authors had hypothesized that affective temperaments represent the subclinical manifestations of mood disorders: in particular, cyclothymic and hyperthymic temperaments have been considered as a subthreshold variant of bipolar disorder. The aim of our study is to test the presence of affective temperaments in a group of Italian patients with major depressive disorder (MDD), and to test the association between cyclothymic temperament and well-established validators of bipolar disorder diagnosis such as age at onset and family history of bipolar disorder. METHODS: Patients with diagnosis of major depressive disorder (DSM-IV-TR) were included in the study. Affective temperaments have been evaluated through the Italian semistructured interview version of the Temperament Evaluation of Memphis, Pisa, Paris and San Diego (TEMPS-I). In order to improve the accuracy of family history and age at onset reports, close family members of the patients were also interviewed. RESULTS: 104 of patients included in the study have completed the temperament interview. 25.5% were diagnosed with a dominant affective temperament. Cyclothymic affective temperament was the most represented in the sample of MDD patients (12.3%); depressive, hyperthymic and irritable temperaments have been detected respectively in 7.5%, 2.8% and 2.8% of patients. Patients with CT showed a significantly lower age at onset of MDD than "pure" MDD patients (31.9 vs. 40.9 years; p=0.049) and higher rates of family history of bipolar disorder in first degree relatives (15.4% vs. 0%; p=0.001). LIMITATIONS: The major limitation of this study was the lack of a group of bipolar depressives, which would have been useful in order to confirm the similarities of age at onset and bipolar family history with cyclothymic MDD. CONCLUSIONS: Our data confirm previous reports in a sample of accurately screened patients with unipolar major depression: we found that patients with a cyclothymic temperament had an earlier age at onset and a higher family history for bipolar disorder than patients without any dominant affective temperament. Further research is needed to ascertain whether patients with "unipolar" cyclothymic MDD respond to mood stabilizers. PMID- 19556008 TI - High prevalence of human rhinovirus C infection in Thai children with acute lower respiratory tract disease. AB - OBJECTIVE: To determine the prevalence of human rhinoviruses (HRV) infections in children with lower respiratory disease in Thailand and monitor the association between species of HRV and clinical presentation in hospitalized paediatric patients. METHOD: Two hundred and eighty-nine nasopharyngeal (NP) suction specimens were collected from hospitalized paediatric patients admitted to King Chulalongkorn Memorial Hospital, Thailand during February 2006-2007. Nucleic acids were extracted from each sample with subsequent amplification of VP4/2 by semi-nested RT-PCR for HRV detection. Other viral respiratory pathogens were also detected by PCR, RT-PCR or real time PCR. Nucleotide sequences of the VP4 region were used for genotyping and phylogenetic tree construction. RESULT: In total, 87 of 289 specimens were positive for HRV indicating an annual prevalence of 30%. Wheezing or asthma exacerbation was the most common clinical presentation observed in infected patients. Sequence analysis and phylogenetic tree showed that 29 (33%) and 8 (9%) specimens belonged to HRV-A and HRV-B, respectively. Most of the HRV positive samples were HRV-C (58%). Moreover, species C was predominantly found in the paediatric population of Thailand in raining season (p<0.05). The frequency of co-infection of HRV-C with other respiratory viral pathogens was approximately 40%. CONCLUSION: HRV-C represents the predominant species and is one of the etiologic agents in acute lower respiratory tract infection, causes of wheezing and asthma exacerbation in infants and young children in Thailand. PMID- 19556010 TI - Pravastatin prevents miscarriages in antiphospholipid antibody-treated mice. AB - Miscarriages in patients with antiphospholipid (aPL) antibodies have been attributed to thrombosis of placental vessels. However, we have shown that inflammation plays a crucial role in fetal injury. We identified tissue factor (TF), the major cellular activator of the coagulation cascade, as a key mediator in inflammation and fetal injury in aPL antibody-treated mice. We found that TF in maternal neutrophils was associated with fetal injury. TF expression in neutrophils contributes to the respiratory burst and subsequent trophoblast oxidative injury and pregnancy loss induced by aPL antibodies. We also analysed how TF contributes to neutrophil activation and trophoblast injury in this model. We showed that neutrophils from aPL antibody-treated mice express protease activated receptor 2 (PAR-2) and that stimulation of this receptor leads to neutrophil activation, trophoblast injury and fetal death. Mice deficient in PAR 2 and treated with aPL antibodies exhibited reduced neutrophil activation and normal pregnancies, indicating that PAR-2 plays an important role in the pathogenesis of aPL antibody-induced fetal injury. In addition, we demonstrated that the statins simvastatin and pravastatin downregulate TF and PAR-2 expression in neutrophils and thus prevent pregnancy loss. In summary, this study shows that TF signaling through PAR-2 mediates neutrophil activation and fetal death in antiphospholipid syndrome, and that statins may be an appropriate treatment for women with aPL antibody-induced pregnancy complications. PMID- 19556011 TI - Complete-cases analysis is appropriate for randomised trials with pre-test-post test designs. PMID- 19556012 TI - Inflammatory cytokines and the nuclear vitamin D receptor are implicated in the pathophysiology of dental resorptive lesions in cats. AB - Dental resorptive lesions (RL) are a common oral disease in cats (Felis catus) associated with pain and tooth destruction. The aetiology of RL in cats is unknown, but inflammation is often seen in conjunction with RL. Vitamin D involvement has been suggested because 1,25-dihydroxyvitamin D (1,25(OH)(2)D) stimulates osteoclastogenesis, through up-regulation of the nuclear vitamin D receptor (nVDR). The aim of this study is to determine the involvement of inflammatory cytokines and the possible role of vitamin D in the pathophysiology of RL using quantitative PCR. We measured the mRNA expression of cytokines with stimulatory (IL-1beta, IL-6, and TNF-alpha) and inhibitory effects (IL-10 and IFN gamma) on osteoclastogenesis, and the mRNA expression of the receptor activator of nuclear factor-kappaB ligand (RANKL), osteoprotegerin (OPG), and nVDR in RL samples. We found increased expression of mRNA levels for inflammatory cytokines and nVDR, but not for RANKL and OPG, in tissue from RL-affected cats compared with tissue from radiological confirmed healthy controls. The mRNA levels of nVDR were positively correlated with mRNA levels of pro-inflammatory (IL-1beta, IL-6, TNF-alpha, and IFN-gamma), anti-inflammatory (IL-10), pro-resorptive (IL-1beta, IL-6, and TNF-alpha), and anti-resorptive (IFN-gamma and IL-10) cytokines in the course of resorptive lesions. These data are consistent with our view that both inflammation and an overexpression of the nVDR are likely to be involved in RL in cats. PMID- 19556013 TI - Ontogeny of IgM-producing cells in the mandarin fish Siniperca chuatsi identified by in situ hybridisation. AB - The ontogeny of IgM-producing cells was studied in juvenile mandarin fish Siniperca chuatsi, an important fish in China's aquaculture sector. The IgM producing cells were localised through in situ hybridisation with a probe complementary to the Ig mu-chain in lymphoid-related tissues, including head kidney, spleen, thymus, intestine and gills. In head kidney, transcripts of Ig mu were first detected at 20days post-hatching (dph) with a few positive signals, and the number of IgM-producing cells increased obviously from 39dph onwards. At 136dph, a large amount of positive cells were observed in the entire organ with clusters of these cells located around the blood vessels. In spleen, IgM producing cells were found from 26dph onwards, followed by an increase until 67dph; clusters of positive cells were also detected around blood vessels at 102dph. In thymus, IgM-producing cells were first observed at 39dph; thereafter, no obvious increase was detected until 78dph. The positive cells in thymus were distributed mainly in the outer zone of thymus. A few IgM-producing cells were still observed in thymus of 1-year-old mandarin fish. IgM-producing cells were not detected in the intestine until 87dph, with several discrete positively stained cells distributed in the lamina propria. IgM-producing cells, scattered mainly in primary gill filaments around blood vessels, were detected in gills from 90dph. As in other teleosts, these results indicated that the head kidney appears to be the primary organ for IgM production in mandarin fish, and IgM producing cells exist in all organs examined in the present study, implying their lymphoid role in fish. In addition, it is suggested that vaccination after 20dph may be much more effective in mandarin fish. PMID- 19556014 TI - Secretory expression of porcine interferon-gamma in baculovirus using HBM signal peptide and its inhibition activity on the replication of porcine reproductive and respiratory syndrome virus. AB - The gene sequence encoding mature porcine interferon-gamma (PoIFN-gamma) fused with a C-terminal 6x histidine tag was cloned into the baculovirus pFastBac Dual vector of the Bac-to-Bac Baculovirus expression system under the control of PH promoter. The authentic signal sequence of porcine interferon-gamma was substituted with the honeybee melittin (HBM) signal sequence, and expressed in insect cells. The recombinant proteins were detected by SDS-PAGE and immunofluorescence assay. The nickel affinity column purified recombinant porcine interferon-gamma with HBM signal peptide (rPoIFN-gammaH) was shown to be a 19kDa protein as confirmed by Western blot analysis. The recombinant PoIFN-gammaH was shown to have cytokine activity, inhibiting the cytopathic effect of vesicular stomatitis virus (VSV) in PK-15 cells at about 1.07x10(6)U/mL. The 2(-7) dilution of the rPoIFN-gammaH in culture supernatant protected the MARC-145 cells from the cytopathic effect caused by 100TCID(50) of porcine reproductive and respiratory syndrome virus. PMID- 19556015 TI - Gold laser tonsillectomy--a safe new method. AB - OBJECTIVES: To determine if Gold laser tonsillectomy is an acceptably safe method of removing tonsils by comparing its complication rates to other established methods, namely Coblation tonsillectomy and cold steel dissection tonsillectomy. STUDY DESIGN: A retrospective review of 748 consecutive patients, ages 2-18, undergoing tonsillectomy at a pediatric teaching institution. METHODS: Tonsillectomy with or without adenoidectomy was performed utilizing either the Gold laser (n=435), Coblation device (n=153), or by cold steel dissection (n=160) between August 2005 and August 2007. Hospital charts were then reviewed to determine the rates of post-tonsillectomy hemorrhage and dehydration requiring hospital admission. RESULTS: In the Gold laser group there were 7 bleeding events (1.61%) and 7 dehydration admissions (1.61%). The Coblation group had 9 bleeding events (5.88%) and 1 dehydration admission (0.65%). The cold steel group had 1 bleed (0.63%) and 2 dehydration admissions (1.25%). The hemorrhage rate associated with Gold laser tonsillectomy was statistically equivalent to cold steel dissection (p=0.3710) and significantly lower than in our Coblation control group (p=0.0286). CONCLUSIONS: Tonsillectomy by means of the Gold laser can be safely performed in the pediatric population. PMID- 19556016 TI - Dexmedetomidine and proprofol in complex microlaryngeal surgery in infants. AB - We describe the case of an infant undergoing endoscopic repair of a laryngeal cleft where the combination of dexmedetomidine and propofol infusions was used as the anesthetic technique. With this regimen, endotracheal intubation was unnecessary during the perioperative period, the procedure lasted approximately 3h, and the child recovered uneventfully. Historically, the techniques used for microlaryngeal surgery involve the use of intermittent endotracheal intubation and insufflation of halogenated anesthetics to the oropharynx. Given the potential benefits of a technique that obviates the need for endotracheal intubation during microlaryngeal surgery and prevents insufflation of halogenated anesthetics in an open environment, the combination of propofol and dexmedetomidine should be considered as a viable and desirable anesthetic option for infants undergoing complex microlaryngeal surgery. PMID- 19556017 TI - Prevention of postoperative atrial fibrillation and/or flutter is another important reason for percutaneous instead of surgical closure of atrial septal defect in adults. PMID- 19556018 TI - Changes in plasma natriuretic peptide levels in patients with atrial fibrillation after cardioversion. AB - The aim of the study was to assess changes in plasma natriuretic peptide (NP) levels after spontaneous or electrical cardioversion (CV) in patients with paroxysmal or persistent atrial fibrillation (AF). METHODS: Patients with paroxysmal or persistent AF with normal left ventricular function and controlled heart rate, referred for electrical CV, were enrolled prospectively. NP concentrations were measured by means of radioimmunoassay method. RESULTS: We studied 23 patients with paroxysmal and 77 with persistent AF, spontaneously or electrical cardioverted to sinus rhythm. The mean plasma NP levels were increased in AF patients, ANP: 249+/-88.3 pg/ml and 258+/-89.7 pg/ml vs 67+/-21.2 pg/ml; (p<0.001) and BNP: 99.6+/-29.8 pg/ml and 82.3+/-33 pg/ml vs 37.5+/-13 pg/ml; in the paroxysmal, persistent, and control group, respectively. The mean ANP and BNP levels decreased after sinus restoration from 249.0+/-88.3 pg/ml to 70.1+/-13 pg/ml, and from 99.6+/-29.8 pg/ml to 37.4+/-8.4 pg/ml, respectively, in the paroxysmal group; from 257.7+/-89.7 pg/ml to 150.0+/-87.2 pg/ml and from 82.3+/ 33 pg/ml to 63.7+/-28.6 pg/ml in the persistent AF group. NP levels remained stable for the next 30 days in the group of patients who remained in sinus rhythm. CONCLUSION: Plasma NP concentrations are significantly reduced or normalized after sinus rhythm restoration in patients with paroxysmal and persistent AF and remain stable within 4 weeks of follow-up, provided that AF does not recur. PMID- 19556019 TI - Double left anterior descending artery arising from right and left sinus of Valsalva in patient with acute coronary syndrome. AB - An aberrant origin of left anterior descending artery (LAD) from right and left sinus of Valsalva is a rare anomaly but clinically important. Coronary angiography (CAG) was performed in patient with acute coronary syndrome, and revealed severe stenosis at the mid of LAD. The distal of LAD was likely to be a total occlusion. It was proved to be a double left anterior descending artery arising from right and left sinus of Valsalva by multidetector-row computed tomography (MDCT) and following CAG. We describe here congenital coronary anomaly case which would be misleading as a total occlusion. PMID- 19556021 TI - The genetic landscape of intellectual disability arising from chromosome X. AB - X-linked mental retardation (XLMR) or intellectual disability (ID) is a common, clinically complex and genetically heterogeneous disease arising from many mutations along the X chromosome. It affects between 1/600-1/1000 males and a substantial number of females. Research during the past decade has identified >90 different XLMR genes, affecting a wide range of cellular processes. Many more genes remain uncharacterized, especially for the non-syndromic XLMR forms. Currently, approximately 11% of X-chromosome genes are implicated in XLMR; however, apart from a few notable exceptions, most contribute individually to <0.1% of the total landscape, which arguably remains only about half complete. There remain many hills to climb and valleys to cross before the ID landscape is fully triangulated. PMID- 19556022 TI - Stereotactic body radiation therapy for early stage non-small cell lung cancer: results of a prospective trial. AB - Patients affected with early stage (IA-IB) non-small cell lung cancer (NSCLC), deemed medically inoperable, are usually treated by conventional 3D-CRT, with poor results in terms of local tumour control and survival. Hypofractionated stereotactic body radiation therapy (SBRT) appears to be a valid alternative option, with high rates of local control and promising survival rates according to recent reported series. We herein report the final results of a prospective phase II trial of SBRT in 62 stage I NSCLC patients, homogeneously treated with three fractions of 15Gy each, given every other day during a 1 week time, up to a total dose of 45Gy; dose was prescribed to the 80%-isodose encompassing planning target volume. Patients were immobilized in a dedicated stereotactic body frame; margins around gross tumour volume were 5mm in the axial plane and 10mm in the longitudinal direction. Median age was 73.7 years. A pathologic confirmation of NSCLC was obtained in 64.5% of patients. Forty-three patients had stage IA and 19 stage IB disease. The majority of patients did not experience any toxicity; mild skin reactions, fatigue, dyspnea/cough or transient thoracic pain were recorded in approximately 10% of patients. With a median follow-up time of 28 months, 2 patients experienced an isolated local relapse, 4 an isolated nodal relapse and 15 a systemic failure. At 3 years, local control rate was 87.8%, cancer-specific survival 72.5%, overall survival 57.1%, with 8 out of 20 non-cancer related deaths. In multivariate analysis, tumour volume was associated with a better outcome. In our series, SBRT was well tolerated and confirmed its efficacy, with local control and survival rates globally superior to those reported using conventional radiotherapy. A longer follow-up is needed in order to establish a correct comparison with surgical series, and to fully ascertain a potential negative impact of SBRT on comorbidities of such a fragile patients population. PMID- 19556023 TI - Sodium induces simultaneous changes in cytosolic calcium and pH in salt-tolerant quince protoplasts. AB - Previous experiments with salt-resistant quince BA29 (Cydonia oblonga cv. Mill.) have shown that this cultivar takes up sodium transiently into the cytosol of shoot protoplasts only in the absence of calcium chloride, or at <1mM calcium chloride. Addition of NaCl > or =100mM to single protoplasts from in vitro cultivated quince in the presence of 1.0mM calcium induced instant changes in the cytosolic concentrations of calcium and protons. These changes were investigated by use of tetra [acetoxymethyl] esters of the fluorescent stilbene chromophores Fura 2 and bis-carboxyethyl-carboxyfluorescein (BCECF), respectively. The cytosolic Ca(2+) dynamics in the protoplasts were dependent on the concentration of NaCl added. The changes in calcium differed in amplitude and final concentration and were correlated in time mainly with changes in pH. Addition of 100-400mM NaCl to the protoplasts caused an oscillating increase in the cytosolic level of calcium, and then a decrease. Addition of mannitol, of equiosmolar concentration to NaCl, did not increase the cytosolic calcium concentration. Moreover, there was no increase in cytosolic calcium when NaCl was added in the presence of calcium binding ethylene glycol-bis(beta-aminoethylether)-N,N,N',N' tetra acetic acid (EGTA), or lantan or verapamil, two inhibitors of plasma membrane calcium channels. Therefore, we conclude that, in salt-resistant quince, sodium induces an influx of calcium into the cytosol by plasma membrane calcium channels, and a simultaneous increase in cytosolic pH. Because these changes were obtained in the presence of 1mM calcium in the medium, they were not due to sodium uptake into the cytosol. PMID- 19556020 TI - Second-hand smoke stimulates lipid accumulation in the liver by modulating AMPK and SREBP-1. AB - BACKGROUND/AIMS: The underlying mechanisms of steatosis, the first stage of non alcoholic fatty liver disease (NAFLD) that is characterized by the accumulation of lipids in hepatocytes, remain unclear. Our study aimed to investigate the hypothesis that cigarette smoke is known to change circulating lipid profiles and thus may also contribute to the accumulation of lipids in the liver. METHODS: Mice and cultured hepatocytes were exposed to sidestream whole smoke (SSW), a major component of "second-hand" smoke and a variety of cellular and molecular approaches were used to study the effects of cigarette smoke on lipid metabolism. RESULTS: SSW increases lipid accumulation in hepatocytes by modulating the activity of 5'-AMP-activated protein kinase (AMPK) and sterol response element binding protein-1 (SREBP-1), two critical molecules involved in lipid synthesis. SSW causes dephosphorylation/ inactivation of AMPK, which contributes to increased activation of SREBP-1. These changes of activity lead to accumulation of triglycerides in hepatocytes. CONCLUSION: These novel findings are important because they point to another risk factor of smoking, i.e., that of contributing to NAFLD. In addition, our results showing that both AMPK and SREBP are critically involved in these effects of smoke point to the potential use of these molecules as targets for treatment of cigarette smoke-induced metabolic diseases. PMID- 19556024 TI - Discovery and validation of urine markers of acute pediatric appendicitis using high-accuracy mass spectrometry. AB - STUDY OBJECTIVE: Molecular definition of disease has been changing all aspects of medical practice, from diagnosis and screening to understanding and treatment. Acute appendicitis is among many human conditions that are complicated by the heterogeneity of clinical presentation and shortage of diagnostic markers. Here, we sought to profile the urine of patients with appendicitis, with the goal of identifying new diagnostic markers. METHODS: Candidate markers were identified from the urine of children with histologically proven appendicitis by using high accuracy mass spectrometry proteome profiling. These systemic and local markers were used to assess the probability of appendicitis in a blinded, prospective study of children being evaluated for acute abdominal pain in our emergency department. Tests of performance of the markers were evaluated against the pathologic diagnosis and histologic grade of appendicitis. RESULTS: Test performance of 57 identified candidate markers was studied in 67 patients, with median age of 11 years, 37% of whom had appendicitis. Several exhibited favorable diagnostic performance, including calgranulin A (S100-A8), alpha-1-acid glycoprotein 1 (orosomucoid), and leucine-rich alpha-2-glycoprotein (LRG), with the receiver operating characteristic area under the curve and values of 0.84 (95% confidence interval [CI] 0.72 to 0.95), 0.84 (95% CI 0.72 to 0.95), and 0.97 (95% CI 0.93 to 1.0), respectively. LRG was enriched in diseased appendices, and its abundance correlated with severity of appendicitis. CONCLUSION: High-accuracy mass spectrometry urine proteome profiling allowed identification of diagnostic markers of acute appendicitis. Usage of LRG and other identified biomarkers may improve the diagnostic accuracy of clinical evaluations of appendicitis. PMID- 19556025 TI - Disequilibrium between admitted and discharged hospitalized patients affects emergency department length of stay. AB - STUDY OBJECTIVE: Most patients are admitted to the hospital through the emergency department (ED), and ED waiting times partly reflect the availability of inpatient beds. We test whether the balance between daily hospital admissions and discharges affects next-day ED length of stay. METHODS: We conducted a cross sectional study of hospitals in metropolitan Toronto, served by a single emergency medical services provider in a publicly funded system. During a 3-year period, we evaluated the daily ratio of admissions to discharges at each hospital and the next-day median ED length of stay in the same hospital by using linear regression. RESULTS: Across hospitals, the daily mean (SD) 50th percentile ED length of stay averaged 218 (51) minutes. As the inpatient admission-discharge ratio increased or decreased, next-day ED length of stay changed accordingly. Compared with ratios of 1.0, those less than 0.6 were associated with an 11 minute (95% confidence interval [CI] 5 to 16 minutes) shorter next-day median ED length of stay; at admission-discharge ratios of 1.3 to 1.4, ED length of stay was significantly prolonged by 5 minutes (95% CI 3 to 6 minutes). Admission discharge ratios on weekends and among medical inpatients had a stronger influence on next-day ED length of stay; effects were also greater among higher acuity and admitted ED patients. CONCLUSION: Disequilibrium between the number of admitted and discharged inpatients significantly affects next-day ED length of stay. Better matching of daily hospital discharges and admissions could reduce ED waiting times and may be more amenable to intervention than reducing admissions alone. The admission-discharge ratio may also provide a simple way of tracking and enhancing hospital system performance. PMID- 19556026 TI - A multicenter evaluation of the ABCD2 score's accuracy for predicting early ischemic stroke in admitted patients with transient ischemic attack. AB - STUDY OBJECTIVE: We evaluate, in admitted patients with transient ischemic attack, the accuracy of the ABCD(2) (age [A], blood pressure [B], clinical features [weakness/speech disturbance] [C], transient ischemic attack duration [D], and diabetes history [D]) score in predicting ischemic stroke within 7 days. METHODS: At 16 North Carolina hospitals, we enrolled a prospective, nonconsecutive sample of admitted patients with transient ischemic attack and with no stroke history, presenting within 24 hours of transient ischemic attack symptom onset. We conducted a medical record review to determine ischemic stroke outcomes within 7 days. According to a modified Rankin Scale Score, strokes were classified as disabling (>2) or nondisabling (< or =2). RESULTS: During a 35 month period, we enrolled 1,667 patients, of whom 373 (23%) received a diagnosis of an ischemic stroke within 7 days. Eighteen percent (69/373) of all strokes were disabling. We were unable to calculate an ABCD(2) score in 613 patients (37%); however, our imputed analysis indicated this did not significantly alter results. The discriminatory power of the ABCD(2) score was modest for ischemic stroke in 7 days (c statistic 0.59), and fair for disabling ischemic stroke within 7 days (c statistic 0.71). Patients characterized as low risk according to ABCD(2) score (< or =3) were at low risk for experiencing a disabling stroke within 7 days, with a negative likelihood ratio of 0.16 (95% confidence interval [CI] 0.04 to 0.64) with missing values excluded and 0.34 (95% CI 0.15 to 0.76) when missing values were imputed. CONCLUSION: Our analysis suggests the best application of the ABCD(2) score may be to identify patients at low risk for an early disabling ischemic stroke. Further study of the ability to determine an ABCD(2) score in all patients is needed, along with validation in a large, consecutive population of patients with transient ischemic attack. PMID- 19556027 TI - Letters, we've got letters... PMID- 19556028 TI - Comparison of the 20-hour intravenous and 72-hour oral acetylcysteine protocols for the treatment of acute acetaminophen poisoning. AB - STUDY OBJECTIVE: To compare outcomes after acute acetaminophen poisoning in 2 large cohorts of patients treated with either the 20-hour intravenous or 72-hour oral acetylcysteine protocol. METHODS: We conducted a retrospective cohort study with historical control comparing patients treated with one of 2 acetylcysteine regimens. Data for the 20-hour group were obtained from a medical record review of patients on whom the 20-hour intravenous protocol was initiated in Canadian hospitals from 1980 to 2005. The 72-hour group consisted of a historical cohort of patients treated in US hospitals with the 72-hour oral protocol from 1976 to 1985. The primary outcome was hepatotoxicity (aminotransferase levels >1,000 IU/L). RESULTS: Of the 4,048 patients analyzed, 2,086 were in the 20-hour group and 1,962 were in the 72-hour group. The incidence of hepatotoxicity was 13.9% in the 20-hour group and 15.8% in the 72-hour group (-1.9% absolute difference; 95% confidence interval [CI] -4.2 to 0.3). The relative risk of hepatotoxicity was lower in the 20-hour group when acetylcysteine was initiated within 12 hours of ingestion. The relative risk was lower in the 72-hour group when acetylcysteine was initiated later than 18 hours after ingestion. There was no significant risk difference between groups when acetylcysteine treatment was started 12 to 18 hours after ingestion. One patient in the 20-hour group received a liver transplant and died because of acetaminophen toxicity compared with no liver transplants and 3 deaths in the 72-hour group. Anaphylactoid reactions to intravenous acetylcysteine were reported in 148 of 2,086 patients (7.1%; 95% CI 6.1% to 8.3%). This study is limited by comparison of 2 separate data sets from different countries and study years. CONCLUSION: The risk of hepatotoxicity differed between the 20-hour and 72-hour protocols according to the time to initiation of acetylcysteine. It favored the 20-hour protocol for patients presenting early and favored the 72-hour protocol for patients presenting late after acute acetaminophen overdose. PMID- 19556029 TI - Emergency department bedside ultrasonographic measurement of the caval index for noninvasive determination of low central venous pressure. AB - STUDY OBJECTIVE: Among adult emergency department (ED) patients undergoing central venous catheterization, we determine whether a greater than or equal to 50% decrease in inferior vena cava diameter is associated with a central venous pressure of less than 8 mm Hg. METHODS: Adult patients undergoing central venous catheterization were enrolled in a prospective, observational study. Inferior vena cava inspiratory and expiratory diameters were measured by 2-dimensional bedside ultrasonography. The caval index was calculated as the relative decrease in inferior vena cava diameter during 1 respiratory cycle. The correlation of central venous pressure and caval index was calculated. The sensitivity, specificity, and positive and negative predictive values of a caval index greater than or equal to 50% that was associated with a central venous pressure less than 8 mm Hg were estimated. RESULTS: Of 73 patients, the median age was 63 years and 60% were women. Mean time and fluid administered from ultrasonographic measurement to central venous pressure determination were 6.5 minutes and 45 mL, respectively. Of the 73 participants, 32% had a central venous pressure less than 8 mm Hg. The correlation between caval index and central venous pressure was 0.74 (95% confidence interval [CI] -0.82 to -0.63). The sensitivity of caval index greater than or equal to 50% to predict a central venous pressure less than 8 mm Hg was 91% (95% CI 71% to 99%), the specificity was 94% (95% CI 84% to 99%), the positive predictive value was 87% (95% CI 66% to 97%), and the negative predictive value was 96% (95% CI 86% to 99%). CONCLUSION: Bedside ultrasonographic measurement of caval index greater than or equal to 50% is strongly associated with a low central venous pressure. Bedside measurements of caval index could be a useful noninvasive tool to determine central venous pressure during the initial evaluation of the ED patient. PMID- 19556030 TI - Optimizing emergency department front-end operations. AB - As administrators evaluate potential approaches to improve cost, quality, and throughput efficiencies in the emergency department (ED), "front-end" operations become an important area of focus. Interventions such as immediate bedding, bedside registration, advanced triage (triage-based care) protocols, physician/practitioner at triage, dedicated "fast track" service line, tracking systems and whiteboards, wireless communication devices, kiosk self check-in, and personal health record technology ("smart cards") have been offered as potential solutions to streamline the front-end processing of ED patients, which becomes crucial during periods of full capacity, crowding, and surges. Although each of these operational improvement strategies has been described in the lay literature, various reports exist in the academic literature about their effect on front-end operations. In this report, we present a review of the current body of academic literature, with the goal of identifying select high-impact front-end operational improvement solutions. PMID- 19556031 TI - Acute metformin overdose: examining serum pH, lactate level, and metformin concentrations in survivors versus nonsurvivors: a systematic review of the literature. AB - STUDY OBJECTIVE: Metformin is known to cause potentially fatal metabolic acidosis with an increased lactate level in both overdose and therapeutic use. No association between mortality and serum pH, lactate level, or metformin concentrations, though intuitive, has yet been described. This systematic literature review is designed to evaluate the association between mortality and serum pH, lactate level, and metformin concentrations in acute metformin overdose. METHODS: We reviewed the literature by using the MEDLINE, EMBASE, CINAHL, and TOXNET databases for cases of metformin overdose with documented mortality data and values of serum pH, lactate level, and metformin concentrations. When available, patient age, patient sex, and whether patients received intravenous sodium bicarbonate therapy or hemodialysis were also analyzed. Cases meeting inclusion criteria were analyzed to determine whether a difference in distribution of nadir serum pH, peak serum lactate level, or peak serum metformin concentrations existed between overdose survivors and nonsurvivors. RESULTS: We identified 10 articles that had 1 or more cases meeting our inclusion criteria. In total, there were 22 cases of metformin overdose (5/22 died) that met inclusion criteria. No intentional overdose patients died whose serum pH nadir was greater than 6.9, maximum lactate concentration less than 25 mol/L, or maximum metformin concentration less than 50 microg/mL (therapeutic range 1 to 2 microg/mL). Intentional overdose patients with a nadir serum pH less than 6.9 had 83% mortality (5/6), those with lactate concentration greater than 25 mmol/L had 83% mortality (5/6), and those with metformin concentration greater than 50 microg/mL had 38% mortality (5/12). Nadir serum pH and peak serum lactate and metformin concentration distributions in survivors and nonsurvivors revealed that survivors had a median nadir pH of 7.30, interquartile range (IQR) 7.22, 7.36; nonsurvivors, a median nadir pH of 6.71, IQR 6.71, 6.73; survivors, a median peak lactate level of 10.8 mmol/L, IQR 4.2, 12.9; nonsurvivors, a median peak lactate level of 35.0 mmol/L, IQR 33.3, 39.0; survivors, a median peak metformin level of 42 microg/mL, IQR 6.6, 67.6; and nonsurvivors, a median peak metformin level of 110 microg/mL, IQR 110, 110. CONCLUSION: No cases of acute metformin overdose meeting the study's inclusion criteria were found in which patients with a nadir serum pH greater than 6.9, peak serum lactate concentrations less than 25 mmol/L, or peak serum metformin concentrations less than 50 microg/mL died. Patients with acute metformin overdose who died had much lower serum pH nadirs and much higher peak serum lactate and metformin concentrations than those who survived. PMID- 19556032 TI - Clinical and ECG effects of escitalopram overdose. AB - STUDY OBJECTIVE: We investigate the clinical effects of escitalopram overdose and determine the risk of QT prolongation and serotonin toxicity. METHODS: A review of escitalopram overdoses to a clinical toxicology unit was undertaken. Patient demographics, details of the ingestion, clinical effects, including evidence of serotonin toxicity, complications (arrhythmias and seizures), ICU admission, and length of stay were obtained. QT and QRS intervals were manually measured on ECGs by using a standardized approach. In a subgroup of 34 prospectively recruited patients, escitalopram was detected in blood from 33 patients. Medians and interquartile ranges (IQR) were reported, and QT versus pulse rate was plotted on a QT nomogram to investigate QT prolongation. RESULTS: Median ingested dose in the 79 presentations was 140 mg (IQR 75 to 260 mg; range 20 to 560 mg), and escitalopram was the only drug ingested or all coingested drugs were nontoxic in 46 cases. Median length of stay for patients receiving clinically important coingestants was 19 hours (IQR 9 to 33 hours) compared with that of patients receiving escitalopram alone (median 12 hours; IQR 7 to 19 hours). Serotonin toxicity occurred in 7 of the 46 escitalopram-alone ingestions (15%) but in only 1 of the 33 patients coingesting other medications. Common features were inducible clonus and hyperreflexia. Central nervous system depression and ICU admission were rare in escitalopram-alone overdoses compared with those in cases with sedative coingestants. Bradycardia (pulse rate <60 beats/min) occurred in 11 cases (14%) and an abnormal QT-HR pair in 11 (14%), which was associated with normal or slow pulse rates. There were no deaths, seizures, or arrhythmias. CONCLUSION: Major manifestations of escitalopram overdose were serotonin toxicity, QT prolongation, and bradycardia. The study suggests a potential for cardiac arrhythmias in escitalopram overdose. PMID- 19556033 TI - Surveillance, control, and prevention of surgical site infections in breast cancer surgery: a 5-year experience. AB - BACKGROUND: We analyzed variations in surgical site infections (SSIs) during 5 years of a prospective surveillance program and investigated possible contributors to SSIs in a cohort of patients who underwent surgery for breast cancer. METHODS: All breast surgeries performed between January 2001 and December 2005 were registered. Patients were followed-up by direct observation for at least 30 days under standardized conditions. The main outcome studied was SSI. A case-control analysis was conducted to identify SSI-associated risk factors and to evaluate SSI variations by means of a control chart. RESULTS: During the study period, a total of 2338 breast cancer surgeries were recorded, and 441 SSIs (18.9%) were diagnosed. SSI frequency varied across the 5-year period, with a sharp decline seen after the introduction of preventive policies. After 2002, 3 out-of-confidence limits of SSIs were observed, 2 related to the use of evacuation systems and 1 associated with a group of rotating residents. Concomitant preoperative chemoradiation (odds ratio [OR]=3.47; 95% confidence interval [CI]=2.51 to 4.80), hematoma (OR=3.05; 95% CI=1.70 to 5.52), age > or = 58 years (OR=1.83; 95% CI=1.27 to 2.65), body mass index > or = 30.8 (OR=1.58; 95% CI=1.14 to 2.18), and duration of surgery > or = 160 minutes (OR=1.73; 95% CI=1.20 to 2.50) were found to be SSI-associated risk factors. CONCLUSIONS: After 5 years of a continuous prospective surveillance program, we were able to decrease the rate of SSIs in patients undergoing breast cancer surgery (from 33.3% in 2000 to 18.9% in 2005), identify SSI-associated risk factors, and improve the quality of care delivered to these patients. PMID- 19556034 TI - A prospective study of the impact of colonization following hospital admission by glycopeptide-resistant Enterococci on mortality during a hospital outbreak. AB - BACKGROUND: This study was designed to investigate the impact on mortality of colonization by glycopeptide-resistant Enterococci (GRE) during hospitalization. METHODS: Between 2004 and 2006, a hospital in Nancy, France, was subject to a GRE van A outbreak. Some 113 patients who had acquired GRE after hospital admission were matched with 113 controls. Basic demographic data, such as sex, age, principal pathology, history of surgery, and presence of associated pathology, were obtained for each case and control. Information on whether or not the case subject was still alive was obtained by searching the hospital mortality database and the civil death register and by phoning the patient's home. Statistical analysis used the Cox proportional hazards model for calculating survival function with SPSS software version 9.1 (SPSS Inc., Chicago, IL). RESULTS: The mean age was 71.2 in the GRE+ group and 70.8 in the control group (P = .80). There was a significant difference between the groups for severity status health (P = .035). The mortality rate was 30.1% in the case group and 19.5% in the control group. Single predictor variable analysis showed a hazard ratio of death in the case group of 4.61 (95% confidence interval [CI]: 2.58-8.28], P = 2 x 10( 7)). The final Cox regression model with multiple predictor variables showed that only GRE presence (OR, 1.63 [95% CI: 1.04-2.57], P = .035) and severity of comorbidity (P = .013) were independently significant predictors of mortality. CONCLUSION: This study shows that the GRE acquisition has a poor prognosis and that this is independent of the other prognostic factors such as age and severity of underlying disease. Survival in GRE+ patients was significantly shorter. PMID- 19556035 TI - Influenza vaccination attitudes and practices among US registered nurses. AB - BACKGROUND: The influenza vaccination rate among US health care personnel (HCP) remains low and may vary by occupational categories. The objective of this study was to explore knowledge, attitudes, and beliefs associated with influenza vaccination in a broad population of registered nurses. METHODS: The study used a cross-sectional mail survey, administered January-March 2006, of 2000 registered nurses in 4 US states. RESULTS: Of the 2000 surveys sent, 1310 (72%) were returned, and 1017 (67%) were eligible for analysis. The majority of respondents (59%) reported receiving influenza vaccine during the 2005-2006 influenza season. The most common reason for being vaccinated was protecting oneself from illness (95%), and the most common reason for not being vaccinated was concern about adverse reactions (39%). Respondents who reported their patient population as high risk related to influenza were more likely to be vaccinated and to agree with statements regarding influenza disease and influenza vaccination of HCP. CONCLUSION: Concerns about adverse reactions and vaccine effectiveness continue to be barriers to influenza vaccination among registered nurses. Those most knowledgeable about influenza vaccination of HCP have higher vaccination rates. Future efforts to improve vaccination rates should include data on vaccine effectiveness and adverse effects, as well as descriptions of high-risk populations. PMID- 19556036 TI - Infection prevention and control in home health care: the nurse's bag. AB - This study evaluates bacterial contamination rates of home health care nurses' bags and the patient care equipment found inside. Nurses' bags--a ubiquitous fomite in the home health care environment--may serve as a potential reservoir for multidrug-resistant organisms. PMID- 19556037 TI - [Domestic violence: a screening question]. AB - AIMS: To assess whether key questions can detect domestic violence (DV), and find out the prevalence in primary care (PC) and women's care (WC). DESIGN: Intervention Study with random assignment. LOCATION: PC and WC clinics (Sant Boi de Llobregat Barcelona). PARTICIPANTS: Systematic random selection of women in PC (not know if battered) and women who came for the first time to the WC service. Age-matched controls were selected. A total of 400 women were interviewed. INTERVENTIONS: Anamnesis was performed and a questionnaire filled in. In the control group (CG) there was one question concerning DV, in the intervention group (IG) there were 6 key questions: relationship of their parents, partner's parents, if the partner had suffered abuse, relationship with her partner, if sexual relations were consensual or got angry if they were not. When DV was detected, asked about type, features and duration. RESULTS: A total of 101(25.3%) cases of violence were detected, 58(29.4%) in IG and 43(21.2%) in CG (odds ratio (OR)=1.55, confidence interval (CI): 0.96-2.51, P=0.06). WC rates were 32.7% and 17.5% (OR: 2.3, CI: 1.2-4.5, P:0.007) and PC 25.3% and 25.8% (OR: 0.97, CI: 0.47 2.02, P=0.5), respectively. Different characteristics were observed depending on whether they were from PC or WC, but were similar between IG and CG, except mental disability, more common in IG. 23.5% suffered from a psychiatric disorder, 15% in non-battered and 44% in the battered, in 68% of these the disorder appeared after the abuse. CONCLUSIONS: Abuse is a Public Health problem. When women visit for the first time it is necessary to use indirect questions to make communication easier, if the woman and the professional know each other, a single question is enough to detect it. PMID- 19556038 TI - A new class of anticonvulsants possessing 6 Hz activity: 3,4-dialkyloxy thiophene bishydrazones. AB - Thirty nine new 3,4-di(substituted)oxy-N(2),N(5)-bis(substituted)thiophene-2,5 dicarbohydrazides were synthesized starting from ethyl thiodiglycolate through multi-step reactions. In the synthetic sequence, 3,4-dihydroxythiophene-2,5 diester (1) was obtained by condensing the ethyl thiodiglycolate with diethyl oxalate. It was derivatized using different alkyl halides to give disubstituted thiophene esters (2-5), which were then converted to corresponding hydrazides (6 9) following usual methods. Finally, these hydrazides, on treatment with various substituted carbonyl compounds underwent smooth condensation to yield target hydrazones (10-13). The new compounds were characterized using FT-IR, (1)H NMR and (13)C NMR, mass spectral and elemental analyses. The anticonvulsant activity of the title compounds was established after intraperitoneal (ip) administration in three seizure models, which include maximal electroshock (MES), subcutaneous pentylenetetrazole (scPTZ) and 6 Hz screens and their neurotoxicity was also evaluated. Compound 11f has emerged as an active compound with no neurotoxicity in this series. Also, the structure-activity relationship of the tested compounds was discussed. PMID- 19556039 TI - Dangers, and benefits of the cytokine mediated response to injury and infection. AB - The inflammatory response is essential for survival in an environment where continuous exposure to noxious events threaten the integrity of the organism. However, the beneficial effects of the response are influenced by factors, which disadvantage individuals within a population. These factors include malnutrition, infection, genotype, gender, pre-existing inflammation, and chronic intoxication. Although the inflammatory response is generally successful in dealing with noxious events, life-long exposure to these events takes its toll on the integrity of the body and becomes apparent as chronic disease, atherosclerosis, organ failure, and frailty. Progress in ameliorating the consequences of lifetime exposure to inflammatory events can only occur if a fuller understanding can be obtained of the factors, which influence the persistence and outcome of the inflammatory response at an individual level. A multitude of studies has shown that specific nutrients, diets, and dietary restriction are able to modulate the inflammatory response in the population as a whole. To advance in this area, precise knowledge is needed of how the disadvantageous factors, mentioned above, affect the individual's response to anti-inflammatory nutrients. PMID- 19556040 TI - Clinical use of navigation based on cone-beam computer tomography in maxillofacial surgery. AB - Image-guidance in maxillofacial surgery is based predominantly on computed tomographic (CT) images. Its main disadvantage is the considerable amount of radiation to which the patient is exposed, and dental metal artefacts. Recently, a new class of devices based on the concept of cone-beam computed tomography (CBCT) has been introduced for maxillofacial imaging, which we have investigated. In a clinical study, the first seven patients to be operated using a navigation system based on CBCT images, were evaluated. In all cases patient to image recording was uneventful and the surgical objective was reached. The guidance given by the navigation system was helpful. CBCT is an alternative to conventional CT, gives a lower dose of radiation, and costs less. Limitations in the quality of the images and the size of the field of view may restrict its use. It is suitable for image-guided surgery using a navigation system as long as the images show enough of the relevant anatomy and pathology. PMID- 19556041 TI - Computer-assisted technique for the design and manufacture of realistic facial prostheses. AB - We describe a technique for the design and fabrication of realistic facial prostheses using three-dimensional optical imaging and computer-aided design and manufacturing (CAD/CAM). A three-dimensional dataset of the facial surface of a patient who needed a facial prosthesis was obtained using a sensing system, and the three-dimensional image of the face was reconstructed. A decreased and thinned virtual prosthesis was designed based on the facial image and converted into a rapid prototyping wax prosthesis using a selective laser sintering technique. The rapid prototyping wax prosthesis was then dipped into melted base plate wax which generated a layer of wax on the internal and external surfaces to record its size and thickness. The modified prosthesis was evaluated and refined, and processed to generate a realistic silicone prosthesis. Three-dimensional data acquisition using the sensing system and computer-assisted design and manufacture of the prosthesis allows us to see a whole face without damaging the soft tissues or causing discomfort to the patient or exposure to radiation. The final prosthesis was of satisfactory size, shape, and cosmetic appearance, matched the malformed area, and suited the patient's requirements. PMID- 19556044 TI - Young man with kidney failure and hemorrhagic interstitial nephritis. PMID- 19556045 TI - Anabolic interventions in ESRD: light at the end of the tunnel? PMID- 19556042 TI - Idiopathic nephrotic syndrome and atopy: is there a common link? AB - Numerous reports during the last 60 years have reported a strong association between idiopathic nephrotic syndrome and atopic disorders. Idiopathic nephrotic syndrome can be precipitated by allergic reactions and has been associated with both aeroallergens (pollens, mold, and dust) and food allergies. Patients with idiopathic nephrotic syndrome also may show increased serum immunoglobulin E (IgE) levels. A review of the literature suggests that although some idiopathic nephrotic syndrome cases may be associated with allergies, evidence that it is a type of allergic disorder or can be induced by a specific allergen is weak. Rather, it is likely that the proteinuria and increased IgE levels in patients with idiopathic nephrotic syndrome are caused by increased levels of interleukin 13 observed in these patients. Recent studies suggest that interleukin 13, a known stimulator of IgE response, may mediate proteinuria in patients with minimal change disease because of its ability to directly induce CD80 expression on the podocyte. PMID- 19556043 TI - Nephrotic syndrome in diabetic kidney disease: an evaluation and update of the definition. AB - BACKGROUND: Nephrotic syndrome is defined as urine total protein excretion greater than 3.5 g/d or total protein-creatinine ratio greater than 3.5 g/g, low serum albumin level, high serum cholesterol level, and peripheral edema. These threshold levels have not been rigorously evaluated in patients with diabetic kidney disease or by using urine albumin excretion, the preferred measure of proteinuria in patients with diabetes. STUDY DESIGN: Diagnostic test study. SETTING & PARTICIPANTS: Adults with type 2 diabetes, hypertension, and urine total protein level greater than 0.9 g/d enrolled in the Irbesartan in Diabetic Nephropathy Trial. INDEX TEST: Baseline measures of proteinuria (total protein and albumin excretion and protein-creatinine and albumin-creatinine ratios). Linear regression to relate measures. REFERENCE TEST: Other signs and symptoms of nephrotic syndrome at baseline (serum albumin < 3.5 g/dL, serum total cholesterol > 260 mg/dL or use of a statin, and edema or use of a loop diuretic); progression of chronic kidney disease during follow-up (doubling of baseline serum creatinine level or requirement for dialysis or kidney transplantation). Logistic regression to relate index and reference tests. RESULTS: In 1,608 participants, total urine protein level of 3.5 g/d was equivalent to urine albumin level of 2.2 g/d (95% confidence interval, 1.4 to 3.5). Of 1,467 participants, 641 (44%) had urine total protein level of 3.5 g/d or greater at baseline, 132 (9%) had other signs and symptoms of nephrotic syndrome at baseline, and 385 (26%) had progression of kidney disease during a mean follow-up of 2.6 years. Areas under the receiver operating curves for measures of proteinuria were 0.80 to 0.83 for other signs and symptoms of nephrotic syndrome and 0.72 to 0.74 for kidney disease progression. Threshold levels for nephrotic-range proteinuria and albuminuria were close to the points of maximal accuracy for both outcomes. LIMITATIONS: Study population limits generalizability; inability to adjust for several variables known to affect serum albumin levels; lack of spot urine samples. CONCLUSIONS: The historical definition of nephrotic-range proteinuria appears reasonable in patients with diabetic kidney disease. Equivalent thresholds for nephrotic-range albuminuria and albumin-creatinine ratio are 2.2 g/d and 2.2 g/g, respectively. PMID- 19556046 TI - Behavioral management of headache triggers: Avoidance of triggers is an inadequate strategy. AB - The standard clinical advice for individuals who suffer from recurrent headaches is that the best way to prevent headaches is to avoid the triggers. This review challenges that advice from a number of perspectives, including: that the advice is given in a theoretical vacuum; it is associated with practical problems; and it is not evidence-based. The review considers cognate literatures on stress, negative affect, and chronic pain that advocate approach/confront strategies over avoidance strategies. It is suggested that advice to avoid triggers could result in maintenance of the capacity of the trigger to precipitate headaches or even a sensitization process whereby tolerance diminishes. As anxiety researchers have investigated extensively the issue of how stimuli acquire and lose their capacity to elicit fear, this literature is explored to draw inferences for headache triggers. The review concludes with suggestions concerning etiology of chronic headache and associated management implications, and directions for future research. It argues that the philosophy of 'avoidance of triggers' should be replaced with 'coping with triggers,' as the latter includes both avoidance and approach/confront strategies involving exposure to triggers. PMID- 19556047 TI - Differences in stature, BMI, and dietary practices between US born and newly immigrated Hmong children. AB - This study investigated how acculturation influences diet, cultural practices related to cooking and food preparation knowledge, and stature and body mass index (BMI) of Hmong children. Focus groups (n=12) were conducted during the spring of 2008 in St. Paul/Minneapolis, Minnesota with those born in the US and 9 13 years old (n=22), those born in the US and 14-18 years old (n=25), and those born in Thailand or Laos, who had lived in the US< or =5 years, and 14-18 years old (n=21). Respondents also completed a survey instrument to assess acculturation level which was evaluated by questions about social connections, language use, and dietary habits. Compared to those born in the US, those born in Thailand or Laos were significantly shorter and leaner. Those born in the US also showed elevated acculturation levels in language use, social connections, and gender-oriented tasks compared to those born in Thailand or Laos. Themes from focus group discussions were mealtime patterns, determinants of food health, future health concerns, and changing cultural traditions. Acculturation, years lived in the US, and birth place may play an important role in stature and BMI, food and physical activity habits, cooking and food preparation knowledge, and perceptions of health. PMID- 19556048 TI - Directed forgetting in direct and indirect tests of memory: seeking evidence of retrieval inhibition using electrophysiological measures. AB - We investigated whether directed forgetting as elicited by the item-cueing method results solely from differential rehearsal of to-be-remembered vs. to-be forgotten words or, additionally, from inhibitory processes that actively impair retrieval of to-be-forgotten words. During study, participants (N=24) were instructed to remember half of a series of presented words (TBR) and to forget the other half (TBF), as indicated by an instruction cue shown shortly after each word. During test, accuracy and reaction time measures from lexical decisions (indirect memory test) followed by recognition-memory judgements (direct memory test) were supplemented with event-related potential (ERP) recordings. Results from the behavioural measures revealed directed forgetting in the recognition memory test but not the lexical-decision test. ERPs obtained during recognition indicated that TBR words elicited a larger parietal old/new effect than TBF words overall, suggesting that remember/forget instructions impaired conscious recollection processes more severely than familiarity processes. Moreover, TBF words that were successfully forgotten elicited less parietal activity than correctly rejected new words (the reversed old/new effect; Nowicka, A., Jednororog, K., Wypych, M., & Marchewka, A. (2009). Reversed old/new effect for intentionally forgotten words: An ERP study of directed forgetting. International Journal of Psychophysiology, 71, 97-102). This was taken to implicate that inhibitory processes likely affected these items. Enhanced negativities for successfully forgotten TBF words relative to new words were observed in the lexical-decision task at early (150-250 ms) and late (800-1000 ms) time windows, suggesting that inhibitory influences disrupt more than just conscious recollection when memory retrieval is tested indirectly. PMID- 19556049 TI - [Retrospective monocentric comparative evaluation by sifting of vein grafts versus nerve grafts in palmar digital nerves defects. Report of 32 cases]. AB - AIM: Palmar digital nerves defects can be treated by conventional nerve grafts or by means of a conduit, such as a vein. We compared a vein graft technique to a nerve graft technique in a retrospective monocentric study. MATERIAL AND METHOD: A surgeon who was not involved in the treatment reviewed blind 15 nerve grafts and 17 vein grafts. The evaluation concerned sensitivity, pain, donor site morbidity, social integration and autoassessment of the benefits by the patient. Data were compacted by a sifting method eliminating bad results. The classical functional scores (British Medical Research Council, Moberg, Chanson, Alnot, Dumontier) were also used. RESULTS: The evaluation was carried out at least 11 months after treatment. Defect was never greater than 30 mm. After sifting, vein grafts appeared less efficient than nerve grafts (41% good results against 73%), except in emergencies (86% good results). CONCLUSION: For defect loss of no more than 30 mm in emergencies, the authors propose to use vein grafting. In other situations, the surgeon must take into account the patient's profile and the hemi pulp concerned, dominant or non-dominant, before opting for a nerve or a vein graft. PMID- 19556050 TI - [Respiratory medicine and the new A/H1N1 flu: from a Mexican point of view]. PMID- 19556051 TI - [Lung cancer staging by endoscopic and endobronchial ultrasound-guided fine needle aspiration]. AB - Endoscopic and endobronchial ultrasound are complementary techniques. When combined, they allow for nearly complete mediastinal staging in lung cancer. Each technique has inherent strengths and weaknesses, but when used simultaneously they become far more powerful, to the extent that mediastinoscopy -a more expensive and invasive procedure- is expected to decline in use as the application of endoscopic and endobronchial ultrasound becomes more widespread. The incorporation of these ultrasound techniques has been shown to lead to fewer thoracotomies, benefiting patients and also society, given that costs are thereby reduced. We reflect on recent developments in the field, discuss current debates, and propose a view of what the future holds in store. PMID- 19556052 TI - [Consideration on the flu that we do not want to call "swine": an European point of view]. PMID- 19556053 TI - Anxiety and pain during dental injections. AB - OBJECTIVES: The aim of this study was to study the relationship between anxiety and pain felt during a dental injection in a sample of 'normal' patients about to undergo 'invasive' dental treatment. METHODS: Duration and intensity of pain during a dental injection were measured within a sample of 247 patients. In addition, data on dental anxiety, fear of dental pain, type of treatment, amount of anesthetic fluid, injection location and the use of surface anesthesia were also collected. RESULTS: Anxious patients felt more pain and of longer duration than less anxious patients. 28% of variance on the duration of pain felt could be accounted for by fear of dental pain, the use of surface anesthesia and gender. For the intensity of pain felt, 22% of variance could be accounted for by anxiety felt for the injection and dental anxiety. CONCLUSIONS: Pain felt during dental injections is dependent on dental anxiety, fear of dental pain, fear for the injection, gender and amount of injection fluid (rather than the use of surface anesthesia). In other words, some patients are expected to feel elevated levels of pain during dental injection and would benefit from extra attention and care from the dentist. PMID- 19556054 TI - Factors affecting nitrate distribution in shallow groundwater under a beef farm in south eastern Ireland. AB - Groundwater contamination was characterised using a methodology which combines shallow groundwater geochemistry data from 17 piezometers over a 2 yr period in a statistical framework and hydrogeological techniques. Nitrate-N (NO3-N) contaminant mass flux was calculated across three control planes (rows of piezometers) in six isolated plots. Results showed natural attenuation occurs on site although the method does not directly differentiate between dilution and denitrification. It was further investigated whether NO3-N concentration in shallow groundwater (<5 m below ground level) generated from an agricultural point source on a 4.2 ha site on a beef farm in SE Ireland could be predicted from saturated hydraulic conductivity (Ksat) measurements, ground elevation (m Above Ordnance Datum), elevation of groundwater sampling (screen opening interval) (m AOD) and distance from a dirty water point pollution source. Tobit regression, using a background concentration threshold of 2.6 mg NO3-N L(-1) showed, when assessed individually in a step wise procedure, Ksat was significantly related to groundwater NO3-N concentration. Distance of the point dirty water pollution source becomes significant when included with Ksat in the model. The model relationships show areas with higher Ksat values have less time for denitrification to occur, whereas lower Ksat values allow denitrification to occur. Areas with higher permeability transport greater NO3-N fluxes to ground and surface waters. When the distribution of Cl- was examined by the model, Ksat and ground elevation had the most explanatory power but Ksat was not significant pointing to dilution having an effect. Areas with low NO3 concentration and unaffected Cl- concentration points to denitrification, low NO3 concentration and low Cl- chloride concentration points to dilution and combining these findings allows areas of denitrification and dilution to be inferred. The effect of denitrification is further supported as mean groundwater NO3-N was significantly (P<0.05) related to groundwater N2/Ar ratio, redox potential (Eh), dissolved O2 and N2 and was close to being significant with N2O (P=0.08). Calculating contaminant mass flux across more than one control plane is a useful tool to monitor natural attenuation. This tool allows the identification of hot spot areas where intervention other than natural attenuation may be needed to protect receptors. PMID- 19556055 TI - Quantifying the sources of ozone, fine particulate matter, and regional haze in the Southeastern United States. AB - A detailed sensitivity analysis was conducted to quantify the contributions of various emission sources to ozone (O3), fine particulate matter (PM2.5), and regional haze in the Southeastern United States. O3 and particulate matter (PM) levels were estimated using the Community Multiscale Air Quality (CMAQ) modeling system and light extinction values were calculated from modeled PM concentrations. First, the base case was established using the emission projections for the year 2009. Then, in each model run, SO2, primary carbon (PC), NH3, NO(x) or VOC emissions from a particular source category in a certain geographic area were reduced by 30% and the responses were determined by calculating the difference between the results of the reduced emission case and the base case. The sensitivity of summertime O3 to VOC emissions is small in the Southeast and ground-level NO(x) controls are generally more beneficial than elevated NO(x) controls (per unit mass of emissions reduced). SO2 emission reduction is the most beneficial control strategy in reducing summertime PM2.5 levels and improving visibility in the Southeast and electric generating utilities are the single largest source of SO2. Controlling PC emissions can be very effective locally, especially in winter. Reducing NH3 emissions is an effective strategy to reduce wintertime ammonium nitrate (NO3NH4) levels and improve visibility; NO(x) emissions reductions are not as effective. The results presented here will help the development of specific emission control strategies for future attainment of the National Ambient Air Quality Standards in the region. PMID- 19556056 TI - Assessment of cytotoxic and cytogenetic effects of a 1,2,5-thiadiazole derivative on CHO-K1 cells. Its application as corrosion inhibitor. AB - This work focuses on the possible use of phenanthro[9,10-c]-1,2,5-thiadiazole 1,1 dioxide (TDZ) as a harmless corrosion inhibitor. TDZ range-dose providing minimum adverse effects to the environment and human health, with satisfactory corrosion inhibiting properties was evaluated. Cytotoxicity and genotoxicity of TDZ at 0.57 12.50 microM concentration range were tested by neutral red, chromosomal aberrations, mitotic index, and colony formation assays. Results showed a significant increase of chromatid-type aberrations for the highest concentration of TDZ assayed (12.50 microM). Additionally, a reduction in the proliferative rate for lower concentrations was detected by the MI assay. We concluded that TDZ should be used at concentrations lower than 1.16 microM. Corrosion assays performed showed good inhibition effect (ca. 50%) at low (0.65 microM) TDZ concentration. Consequently, our results indicated that TDZ induced a time- and dose-dependent genotoxic and cytotoxic response on CHO-K1 cells. Short assays should be complemented with long exposure tests to simulate chronic contact with TDZ since lower threshold levels may be found for shorter exposures and a wrong safety range could be determined. PMID- 19556057 TI - The role of cation exchange in the sorption of cadmium, copper and lead by soils saturated with magnesium. AB - The displacement of Ca(2+), Mg(2+), K(+) and Al(3+) from the A and Bw or Bt horizons of two soils developed over serpentinized amphibolites when equilibrated in Cu(2+), Cd(2+) or Pb(2+) solutions was determined, together with the concomitant sorption of the heavy metal. The contributions of Mg(2+) to the effective cation exchange capacities of the A and Bt horizons of the Endoleptic Luvisol were 57% and 94%, respectively, and its contributions to those of the A and Bw horizons of the Mollic Cambisol were 70% and 77%, respectively. In all four horizons, cation exchange, chiefly with Mg(2+) and Ca(2+), was the process chiefly responsible for sorption of Cd(2+), Cu(2+) and Pb(2+). Al(3+) and K(+) were hardly implicated, especially in the case of Cd(2+). PMID- 19556058 TI - Exposure to contaminated sediments during recreational activities at a public bathing place. AB - More and more time is spent on recreational activities, but few risk assessments focus specifically on these situations and exposure factor data are often scarce. To assess exposure to contaminants at a public bathing place in an urban environment, we have compiled literature data, conducted observation studies, and analyzed water and sediment samples. The levels of anthropogenic contaminants are high in urban environments and traffic frequently plays an important role. In this study, to characterize variability and uncertainty, the deterministic exposure calculations for metal pollutants were supplemented by a probability bounds analysis for the polycyclic aromatic hydrocarbons (PAH). The results from these calculations show that oral intake is the major exposure route for metals, while skin absorption, with present assumptions, is more important for the PAH. The presently measured levels of contaminants, at this public bathing place, cannot be anticipated to cause any significant adverse influence on public health. This assessment methodology is easy to adapt and can be used routinely in other situations with more heavily contaminated surface sediments and lake water. PMID- 19556059 TI - A new approach to predict the condensed phase heat of formation in acyclic and cyclic nitramines, nitrate esters and nitroaliphatic energetic compounds. AB - This paper presents a new approach to predict the condensed phase heat of formation in important classes of energetic compounds including acyclic and cyclic nitramines, nitrate esters and nitroaliphatics. The condensed phase heat of formation has been expressed by a combination of a core heat of formation, which is a function of elemental composition of the energetic compound, and correction functions. The correction functions for some energetic compounds can have decreasing or increasing effects on the basis of thermodynamic stability. It has been shown that this model can offer reliable predictions for the above energetic compounds compared to the calculated outputs of the best recent empirical and complex quantum mechanical methods. PMID- 19556060 TI - Pieces of a puzzle: Permeability, proinflammatory pathways and pain? PMID- 19556061 TI - Isolation of Toxoplasma gondii from the brain of a dog in Australia and its biological and molecular characterization. AB - Toxoplasma gondii was isolated from the brain of a young dog for the first time in Australia. The identity of the parasite was confirmed by PCR, Western blotting, electron microscopy and cat bioassay. Genotyping of the isolate (TgDgAu1) was determined by PCR-RFLP markers that showed it to be a Type II strain. Western blotting demonstrated the presence of IgM antibodies to T. gondii suggesting the bitch was probably infected during pregnancy and the T. gondii was transmitted to the pups congenitally. We believe this represents the first description of a natural case of congenital transmission of T. gondii in the dog. PMID- 19556062 TI - Foreword to the 22nd WAAVP conference. PMID- 19556063 TI - The comparative efficacy of four anthelmintics against a natural acquired Fasciola hepatica infection in hill sheep flock in the west of Ireland. AB - The aim of this study was to evaluate the efficacy of four different anthelmintics against F. hepatica in a naturally infected hill sheep flock in the west of Ireland. In a randomised trial, 138 ewes were divided into four groups. On day 0 each group was dosed with one of four anthelmintics: triclabendazole, closantel, oxyclozanide or nitroxynil. Faecal samples were obtained from each animal per rectum on the day of treatment and again at 7, 14, 21 and 56 days post treatment. The number of F. hepatica eggs per gram of faeces was determined using the sedimentation technique and the efficacy of each anthelmintic was calculated in terms of the percentage reduction in egg count at each time point. The results for closantel, oxyclozanide and nitroxynil indicate that these drugs are effective with faecal egg count being reduced by 100% by day 14 post-treatment. However, the results for triclabendazole group yielded lower efficacy levels, with faecal egg count reductions of between 49% and 66% based on arithmetic means, over the period 7-56 days post-treatment. These results are highly indicative of triclabendazole resistant F. hepatica in sheep on this farm. PMID- 19556064 TI - Development of an indirect ELISA test using a purified tachyzoite surface antigen SAG1 for sero-diagnosis of canine Toxoplasma gondii infection. AB - The aim of this study was to use affinity purified 30 kDa Toxoplasma gondii surface antigen (SAG1) in an indirect ELISA and to compare it with an existing indirect fluorescent antibody test (IFAT). 245 serum samples were collected from dogs from three adjacent Iranian provinces. IFAT examination revealed positive results in 73 dogs, with titers ranging from 1:16 to 1:1024. A suitable ELISA cut off was determined by ROC analysis in comparison with IFAT. Results showed a relative sensitivity and specificity of 94.52% and 93.60%, respectively, for a cut-off value of 0.790 in the ELISA. No cross-reactivity was detected between antibodies against T. gondii and a closely related protozoan parasite, Neospora caninum, using this newly developed ELISA test. PMID- 19556065 TI - Sheep helminth parasitic disease in south eastern Scotland arising as a possible consequence of climate change. AB - The climate in the UK is changing, with a trend towards increased rainfall in the autumn and winter and warmer average temperatures throughout the year. There has also been a 4-week extension of the herbage growing season over the past 40 years. These changes may have implications for the epidemiology of sheep helminth parasites. Here, we describe production-limiting disease outbreaks caused by Haemonchus contortus, Nematodirus battus, Teladorsagia circumcincta and Fasciola hepatica in sheep flocks in south eastern Scotland. The occurrence and timing of these disease outbreaks could not have been predicted in this region highlighting changes in the epidemiology of helminth infections from the patterns historically described. These cases are used to introduce discussion regarding the potential effects of climate change on the epidemiology of helminth parasites and the implications for sheep farming in the UK. PMID- 19556066 TI - Solitary and social heavy drinking, suicidal ideation, and drinking motives in underage college drinkers. AB - In college students, solitary heavy drinking (i.e., while alone) is associated with depression and with higher rates of drinking problems than heavy drinking in social contexts. This study explored the relationship among heavy episodic drinking context, suicidal ideation, and drinking motives among underage college drinkers (n=91) with a history of passive suicidal ideation. Participants completed measures of depression, suicidal ideation, alcohol consumption and problems, and drinking motives. Multiple regression analyses revealed that suicidal ideation, but not depression, was significantly related to solitary heavy drinking. Neither was related to social heavy drinking. Enhancement motives for drinking, but not other drinking motives (i.e., social, conformity, drinking to cope), were significantly associated with social heavy drinking. In contrast, only drinking to cope was associated with solitary heavy drinking. These findings suggest that greater suicidal ideation is associated with greater frequency of becoming intoxicated while alone, and that this drinking is motivated by attempts to cope. Solitary heavy drinking is a potentially dangerous coping strategy for an individual experiencing suicidal ideation. PMID- 19556068 TI - Pregnancy outcomes for women with epilepsy and bipolar disorder could be improved with intraventricular or intrathecal medication administration. PMID- 19556067 TI - Stress-induced microglial activation may facilitate the progression of neurodegenerative disorders. AB - Microglial activation has been associated with various clinical disorders. In particular, activated microglia have been reported in various brain regions, including the hippocampus, substantia nigra, striatum, and cerebral cortex, in which significant neurodegeneration occurs in various neurodegenerative disorders. Our previous study demonstrated that acute stress, restraint combined with water immersion, substantially induces massive microglial activation in the brain, particularly in the hippocampus, hypothalamus, thalamus, and periaquaductal gray. Furthermore, several studies have also demonstrated that exposure of animals to stress induces microglial activation in the hippocampus and striatum, and also induces the proliferation of microglial cells. Although stress has been shown to contribute to the neurodegenerative changes in the brain, the precise mechanism of this effect remains to be elucidated. Here, we propose that stress-induced microglial activation may be involved in the progression of neurodegenerative changes. Reduction of stress-induced microglial activation may prevent further progression of neurodegenerative disorders. PMID- 19556069 TI - Phantom limb pain--a phenomenon of proprioceptive memory? AB - Despite the amount of research that has been conducted on phantom limb pain (PLP), the etiology of the condition remains unknown, and treatment options are limited. After an individual loses a limb, the brain continues to detect the presence of the missing limb even though it is no longer attached to the body, likely through proprioceptive signals. The majority of patients with amputations either report the feeling of volitional control over their phantom or a phantom limb that is frozen in a specific position. Many patients also experience PLP. Here we propose a new theory, termed "proprioceptive memory," which may explain some of the unique experiences amputees encounter. We also suggest that memories of the limb's position prior to amputation remain embedded within an individual's subconscious, and pain memories that may be associated with each limb position contribute not only to PLP, but to the experience of a fixed or frozen limb. We suspect that there are memory networks for pain--and other sensations, either positive or negative--that are associated with each limb position, and propose that these memories evolved to protect our bodies from repeated injury. A discussion of mirror therapy as a treatment option for PLP is also provided, as well as an explanation for the efficacy of mirror therapy. The paper offers a unique insight into how and why amputees experience these unusual phenomena. PMID- 19556070 TI - Mechanism of increased mortality in hemodialysed patients with periodontitis. AB - Periodontitis significantly decreases survival in hemodialysed patients with end stage renal disease (ESRD). Periodontitis-related microorganisms spreading into the bloodstream are thought to impair blood rheological parameters - for example, increasing whole blood viscosity, aggregating blood elements, and decreasing blood flow - and thereby significantly accelerate systemic or local diseases, impairing survival. We discuss the ability of a prototypical pathogenic anaerobic polybacterial consortium to modulate and interfere with host immune responses and to enzymatically degrade host proteins, to bind to and cleave extracellular matrix proteins, to invade intercellularly as well as intracellularly, to promote vascular permeability, to disrupt polymorphonuclear leukocyte function, to cleave complement, and to degrade IgG heavy chains. To further elucidate these phenomena, studies involving detecting microorganism byproducts and monitoring blood rheological parameters are necessary. PMID- 19556071 TI - Genetic overlap between polycystic ovary syndrome and bipolar disorder: the endophenotype hypothesis. AB - Polycystic Ovary Syndrome (PCOS) is a polygenic disorder caused by the interaction of susceptible genomic polymorphisms with environmental factors. PCOS, characterized by hyperandrogenism and menstrual abnormalities, has a higher prevalence in women with Bipolar Disorder (BD). Theories explaining this high prevalence have included the effect of PCOS itself or the effect of drugs such as Valproate, which may cause PCOS either directly or indirectly. Incidentally, metabolic abnormalities are observed in both bipolar and PCOS patients. Endophenotypes such as insulin resistance, obesity, and hyperglycemia are common among BD and PCOS patients, suggesting some degree of pathophysiological overlap. Since both BD and PCOS are complex polygenetic diseases, the endophenotype overlap may be the result of common genetic markers. This paper postulates that shared clinical endophenotypes between PCOS and BD indicate common pathophysiological platforms and will review these for the potential of genetic overlap between the two disorders. PMID- 19556072 TI - PDT combined with Intravesical BCG instillation would form an autovaccine for bladder cancer? AB - Bladder cancer is the second most common urologic malignancy after prostate cancer. Intravesical BCG as a treatment of superficial bladder cancer (SBC) has been used by urologists for over 30 years. Recently, photodynamic therapy (PDT), which uses a red laser and a photosensitive drug to destroy cancer cells, has been showed encouraging results in SBC treatment. However, BCG and PDT are applied to treatment of SBC alone. Currently, cancer vaccines are made in vitro. Several studies confirmed that tumour cells treated in vitro by PDT can be used for generating potent cancer vaccines, which were more effective than other modes of creating whole tumor vaccines, i.e., UV or ionizing irradiation [Gollnick SO, Vaughan L, Henderson BW. Generation of effective antitumor vaccines using photodynamic therapy. Cancer Res 2002;62:1604-8]. Moreover, BCG is a pleiotropic immune stimulator oriented to cellular immunity. We thought that: after PDT destroyed targeted tumor cells on a large scale, Intravesical BCG could elicit and amplify the immune responses, which would directly form an in situ autovaccine in vivo against the primary tumor and metastases at distant sites. In this paper, we propose that the combination of Intravesical BCG and PDT would be a promising new modality for bladder cancer. PMID- 19556073 TI - Prognostic significance of fascin expression in extrahepatic bile duct carcinomas. AB - Extrahepatic bile duct (EBD) carcinoma is an aggressive cancer with a poor prognosis. Fascin is an actin-bundling protein with roles in forming cell protrusions and mesenchymal and neuronal cell motility. Many human neoplasms up regulate fascin. High fascin expression is thought to be a poor prognostic factor in some cancers; however, few data are available for the role of fascin in EBD carcinoma. We investigated fascin immunoreactivity in EBD carcinoma and tested it for correlations between fascin expression and clinicopathological parameters. Conventional tissue sections of 114 cases of EBD carcinomas were immunohistochemically analyzed for fascin expression. Fascin expression was tested by cytoplasmic staining. Negative, weak positive, and strong positive fascin staining was observed in 49 (43.0%), 32 (28.1%), and 33 cases (28.9%), respectively. Fascin expression in EBD carcinomas was significantly correlated with histological grade (P<0.0001), primary tumor (T) (P=0.002), TNM stage (P=0.036), lymphatic invasion (P=0.048), venous invasion (P=0.024), and adjacent organ invasion (P<0.0001). High fascin expression was a significant poor prognostic factor (P=0.0001) in EBD carcinoma. High fascin expression (P=0.004) and TNM stage (P=0.001) in EBD carcinoma were independent adverse prognostic factors. High fascin expression is significantly correlated with aggressive tumor phenotype in EBD carcinoma and is an independent poor prognostic factor in EBD carcinoma. PMID- 19556074 TI - Loss of anterior translation of the distal humeral articular surface is associated with decreased elbow flexion. AB - PURPOSE: The normal anterior translation of the articular surface of the distal humerus with respect to the humeral diaphysis facilitates elbow flexion. We hypothesize that there is a correlation between anterior translation of the distal humeral articular surface and flexion after open reduction and internal fixation (ORIF) of a fracture of the distal humerus. METHODS: Two independent observers evaluated 141 lateral radiographs of patients more than 6 months after fracture of the distal humerus and 155 lateral radiographs of patients without injury of the distal humerus. The distance between the most anterior point of the distal humerus articular surface, perpendicular to the humeral shaft, from the anterior border of the distal part of the humeral diaphysis, was measured on lateral radiographs as a percentage of the width of the humeral shaft. RESULTS: The technique of measuring anterior translation of the distal humeral articular surface had good intra- and interobserver reliability. The most anterior point of the distal humeral articular surface lies an average of 11.7 mm (range, 6.8 to 17.0 mm) in front of the most anterior border of the humeral shaft in normal distal humeri, which represents 62% of the humeral shaft diameter (range, 33% to 91%). There was a limited but significant correlation between flexion and anterior translation as a percentage of the humeral shaft diameter in distal humeri after fracture that was maintained in multivariable statistical models. CONCLUSIONS: Using a reproducible technique for measuring anterior translation of the distal humerus, there was a correlation between anterior translation of the distal humeral articular surface and elbow flexion after ORIF. Although the weakness of the correlation emphasizes that limitation of elbow flexion after ORIF of a distal humerus fracture is multifactorial, reduced anterior translation of the distal humeral articular surface might be a contributing factor. TYPE OF STUDY/LEVEL OF EVIDENCE: Prognostic IV. PMID- 19556075 TI - Arthroscopically assisted reattachment of avulsed triangular fibrocartilage complex to the fovea of the ulnar head. AB - Triangular fibrocartilage complex (TFCC) insertion into the fovea of the distal ulna plays a crucial role in stabilizing the distal radioulnar joint. Consequently, surgical reattachment against avulsion of the foveal TFCC insertion is required to stabilize the distal radioulnar joint. However, because of technical difficulties, no arthroscopic procedure for such a lesion has currently been established. We present a new technique for arthroscopic reattachment of the avulsed TFCC into the fovea. An osseous tunnel 2.9 mm in diameter is created from the ulnar neck to the foveal surface. Under arthroscopic guidance, a nonabsorbable suture passed into a 21-gauge needle is placed into the TFCC through the osseous tunnel. The avulsed portion of the TFCC is anchored to the fovea by means of a repair suture passed through the TFCC. To achieve normal tension of the TFCC, the suture is tied onto the periosteum around the proximal entrance of the osseous tunnel. Our arthroscopic technique is relatively simple and has great advantages for progressive healing at the attachment site between the TFCC and the fovea. PMID- 19556076 TI - Effect of sensory re-education after low median nerve complete transection and repair. AB - PURPOSE: To compare the sensory results of patients with low median nerve complete transection and repair, with and without sensory re-education. METHODS: We studied 40 patients, aged 20 to 32 years, with low median nerve complete transection. Primary epineural repair using 8-0 single-strand sutures was done in all patients. Hands were immobilized in a splint for 4 weeks, followed by physical therapy for 1 month. At a mean of 3.5 months (range, 3-4 months) after surgery, when vibration sense (pallesthesia), using the 256-cycles-per-second tuning fork, was perceived at the fingertips of the 3.5 radial fingers innervated by the median nerve, the patients were randomly assigned to 2 equal groups: group A patients were rehabilitated with a sensory re-education program, and group B patients had no further treatment. Clinical evaluation at 18 months after surgery (range, 17.5-18.5 months) included locognosia (the ability to localize touch), the static and moving 2-point discrimination tests, and the Moberg pick-up test. RESULTS: All patients were included in the postoperative evaluation. Static and moving 2-point discrimination were not statistically significant between groups. Locognosia was significantly improved in group A, and a statistical trend was identified regarding the Moberg pick-up test in group A compared to group B. CONCLUSIONS: Sensory re-education appeared to have significant value only in re education of locognosia at 18 months after low median nerve complete transection and repair. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic II. PMID- 19556077 TI - Clinical outcomes of surgical release among diabetic patients with carpal tunnel syndrome: prospective follow-up with matched controls. AB - PURPOSE: To compare the clinical outcome after carpal tunnel release in diabetic and nondiabetic patients. METHODS: We evaluated a prospective, consecutive series of 35 diabetic patients (median age, 54 years; 15 with type 1 and 20 with type 2 diabetes) with carpal tunnel syndrome, who were age- and gender-matched with 31 nondiabetic patients (median age, 51 years) having idiopathic carpal tunnel syndrome. Exclusion criteria were other focal nerve entrapments, cervical radiculopathy, inflammatory joint disease, renal failure, thyroid disorders, previous wrist fracture, and long-term exposure to vibrating tools. Participants were examined independently at baseline (preoperatively) and 6, 12, and 52 weeks after surgery, including evaluating sensory function (Semmes-Weinstein), motor function (abductor pollicis brevis muscle strength and grip strength), pillar pain, cold intolerance, and patient satisfaction. RESULTS: The number of patients with normal sensory function (pulp of index finger) increased notably in both patient groups from baseline (diabetic patients, 7 of 35; nondiabetic patients, 10 of 31) compared with the 52-week follow-up (diabetic patients, 25 of 35; nondiabetic patients, 24 of 31). Grip strength decreased temporarily at 6 weeks but recovered completely after 12 weeks. At the 52-week follow-up, mean grip strength (95% confidence interval) had improved significantly in both patient groups (diabetic patients: 3.0 kg [-0.3 to 6.2], nondiabetic patients: 3.4 kg [0.2 to 6.6]). Pillar pain correlated significantly with grip strength at the 6 week follow-up (r(s) = -0.41 to -0.54 [p < .05]). The number of patients reporting cold intolerance decreased over time (diabetic patients, 22 of 35 to 19 of 35; nondiabetic patients, 18 of 31 to 8 of 31), but decreased markedly less for the diabetic patients. Level of patient satisfaction was equal between groups. Comparing type 1 and type 2 diabetic patients, no important difference was noted on any test variables. CONCLUSIONS: Patients with diabetes have the same beneficial outcome after carpal tunnel release as nondiabetic patients. Only cold intolerance demonstrated a lesser extent of relief for diabetic patients. TYPE OF STUDY/LEVEL OF EVIDENCE: Prognostic I. PMID- 19556078 TI - The effect of surface treatment using hyaluronic acid and lubricin on the gliding resistance of human extrasynovial tendons in vitro. AB - PURPOSE: To investigate the effects of tendon surface treatment using hyaluronic acid (HA) and lubricin on the gliding resistance of human extrasynovial palmaris longus (PL) tendon in vitro. METHODS: Thirty-two fresh-frozen cadaver human fingers and 16 ipsilateral PL tendons were used. Each PL tendon was divided into 2 pieces, which were randomly assigned into 4 experimental groups. After the gliding resistance of the normal PL tendon segments were measured, the tendons were treated with either saline, carbodiimide derivatized (cd) gelatin and HA (cd HA gelatin), cd gelatin with lubricin added (cd gelatin plus lubricin), or cd-HA gelatin plus lubricin. After treatment, tendon gliding resistance was measured during up to 1000 cycles of simulated flexion and extension motion. RESULTS: The gliding resistance of the PL tendons in the cd-HA gelatin, cd gelatin plus lubricin, and cd-HA gelatin plus lubricin groups was significantly lower than that of the saline-treated control after 1000 cycles. The gliding resistance in these treatment groups decreased within the first 50 cycles and then increased at a much more gradual rate over the 1000 cycles, with the cd-HA gelatin plus lubricin group being most stable. CONCLUSIONS: The results suggest that tendon surface treatment using HA and lubricin can improve the gliding of human PL tendon in vitro. If validated in vivo, tendon surface treatment has the potential to improve the gliding ability of tendon grafts clinically. PMID- 19556080 TI - Isolation of spirochetes of genus Treponema from pigs with ear necrosis. AB - Various ear lesions, often caused by ear biting, are common in pigs. Some herds have a high frequency of ear necrosis, a syndrome characterized by necrotic lesions along the rim of the pinna, often bilateral and sometimes resulting in loss of the entire ear. In samples from such lesions spirochetes have been observed microscopically but never isolated or identified. In this study two herds with periodic outbreaks of ear necrosis among weaners were investigated. Samples were collected from ear lesions and from the gingiva of the pigs. Spirochetes were observed in silver stained histological sections and by phase contrast microscope in scrapings from the necrotic lesions. From an ear lesion a pure spirochete isolate was obtained and identified as a yet unnamed species of genus Treponema, closely related to spirochetes found in digital dermatitis in cattle. From the oral samples two pure isolates were obtained. One of these isolates was identified as the same species as in the ear lesion and one as Treponema socranskii. Species identification was based on 16S rRNA gene sequences. PMID- 19556079 TI - Comparison of antigenic proteins from Lactococcus garvieae KG- and KG+ strains that are recognized by olive flounder (Paralichthys olivaceus) antibodies. AB - Lactococcus garvieae is an important etiological agent of lactococcosis in various fish species including olive flounder (Paralichthys olivaceus). In this study, proteomic and immunoproteomic analyses were employed to compare the antigenic profiles of strains KG9408, MS93003, and NSS9310 strains of L. garvieae. Proteomic analysis using two-dimensional gel electrophoresis (2-DE) revealed differences in five protein spots among the different L. garvieae strains. In immunoproteomic analysis, there was a significant difference in the 2 DE immunoblot profiles of the L. garvieae strains using sera collected from fish surviving infection with either L. garvieae strains KG9408 or NSS9310. These sera reacted with 8 and 7 unique antigenic protein spots, respectively. Heat shock protein (HSP) 70 and DNA-directed RNA polymerase were among the specific antigens recognized by the anti-NSS9310 serum. In addition, the anti-NSS9310 and anti KG9408 olive flounder sera reacted with 25 common antigenic protein spots of all the L. garvieae strains, which included elongation factor (EF)-Tu, arginine deiminase (AD), inosine-5'-monophosphate dehydrogenase (IMPD), glyceraldehyde-3 phosphate dehydrogenase (GAPDH), phosphomannomutase (PMM), L-lactate dehydrogenase (L-LDH), 6-phosphofructokinase and UDP-galactose 4-epimerase (UDP galactose). Based on the present results, the 8 antigens recognized by the anti KG9408 serum and the 25 common antigens recognized by both sera may serve as potential markers for developing an effective vaccine against this bacterium. PMID- 19556081 TI - In vitro penetration of swine oocytes by homologous spermatozoa: Distinct systems for gamete's co-incubation and oocyte's cryopreservation. AB - In vitro penetration (IVP) of swine oocytes by homologous spermatozoa was evaluated in two experiments using four boars as semen donors. In experiment 1, the IVP rate and the number of penetrating spermatozoa (PSP) were compared using three co-incubation systems for vitrified oocytes and fresh sperm: (1) 35mL petri dishes in a CO(2) incubator, (2) 35mL petri dishes in bags (submarine system) and (3) glass flasks partially submerged in water bath with the same gas mixture used for the bag system. Mean PSP was 8.2+/-10.1 and the IVP rate was 90.5%. The PSP differed across all systems (P=0.0006): 15.5+/-0.5 for flasks, 6.3+/-0.4 for CO(2), and 3.9+/-0.4 for bags. The IVP rate for flasks (95.0%) was greater (P=0.01) than for CO(2) and bags (90.8% and 85.0%, respectively), but it did not differ between flasks and CO(2) for three boars (P>0.05). In experiment 2, co incubation was done as described for glass flasks in experiment 1. The IVP rate and PSP were compared for cryopreserved oocytes: either vitrified in open pulled straws (OPS), or frozen in cryotubes. Mean PSP was 5.4+/-6.5 and IVP rate was 89.6%. Both PSP and IVP rate were greater (P<0.0001) for oocytes frozen in cryotubes (7.0+/-0.3% and 95.8%, respectively) than those frozen in OPS (3.7+/ 0.3% and 83.4%, respectively), with no differences found for three boars (P>0.05). In summary, successful IVP of swine oocytes by homologous spermatozoa can be achieved using gametes incubated in glass flasks and oocytes frozen in cryotubes. PMID- 19556082 TI - Acute encephalopathy with biphasic seizures and late reduced diffusion associated with hemophagocytic syndrome. AB - We reported a girl with HHV-6 infection associated with both acute encephalopathy with biphasic seizures and late reduced diffusion, and hemophagocytic syndrome. She had a prolonged convulsion after a one-day history of febrile illness. Cerebrospinal fluid or brain CT showed no abnormalities on admission and her consciousness was recovered on the next day. However, a prolonged seizure and deterioration of consciousness appeared on the sixth day of illness. Diffusion weighted images revealed marked reduction of water diffusion in the bilateral frontal areas. HHV-6 infection was virologically proven by polymerase chain reaction. She was treated with gamma-globulin, steroid pulse therapy, and brain hypothermia. In addition, decrease in white blood cells and platelet counts, and elevation of liver enzymes and ferritin were noted on the fourth day of illness. Hemophagocytic macrophages were revealed by bone marrow aspiration on the sixth day. Her hematological and blood chemistry abnormalities recovered gradually after steroid pulse therapy. An elevation of interleukin-6, -8, and -10, and tumor necrosis factor in the serum and that of interleukin-4, -6, and-8 in the cerebrospinal fluid were observed at the onset of a late seizure. These facts suggested that hypercytokinemia will be related to the pathogenesis of acute encephalopathy of our patient. PMID- 19556083 TI - [Should all old hypertensive subjects have Mini-Mental State Examination?]. AB - Hypertension constitutes a recognized risk factor of vascular dementia but also of Alzheimer-type dementia. Various longitudinal studies showed that midlife blood pressure level is one of the factors conditioning the onset of dementia syndrome in late life. The high risk of dementia is linked to leukoaraiosis, vascular rigidity, microcirculation disorders, oxidative stress, blood pressure fluctuations including orthostatic hypotension and strokes, all of those being associated with hypertension. Numerous clinical trials showed the positive effect of effective treatment of hypertension on the prevention of cognitive disorders and dementias. Thus, screening and early management of dementia and cognitive decline, in particular in the hypertensive subject, are essential. The Mini Mental State Examination (MMSE) is a major first-intention screening test because it allows a full assessment of cognitive aptitudes. If cognitive decline is suspected and the MMSE score is considered to be abnormal, the elderly subject must be sent to a specialist or a referent memory centre; the MMSE is only a first stage in the diagnostic reasoning. MMSE should be included in the follow-up of all hypertensive elderly subjects and should be performed once a year by the general practitioner. PMID- 19556084 TI - [Hepatic transplantation]. AB - The first liver transplantation (LT) was conducted in 1963. After a two-decade development phase with the improvement of surgical and anesthesia-resuscitation techniques, and a better control of allograft rejection, LT has become the benchmark treatment for the majority of end-stage liver diseases. Since the 1980s, indications of LT have gradually expanded and the current main indications in adults are hepatocellular carcinoma at an early stage (15 to 30% of indications), C cirrhosis (15 to 40% of indications) and alcoholic cirrhosis (20 to 25% of indications). Five thousand LTs are performed yearly in Europe, including 1000 LTs in France, with, over the 2000-2006 period, survival rates of 86, 75 and 68% at one, five and 10 years, respectively. Several advances have accompanied the increasing number of indications and these excellent results: (a) the development of more specific immunosuppressive drugs to prevent rejection, the incidence of which is currently less than 20%: tacrolimus, mycophenolic acid, anti-IL2 receptor antibodies, mTOR inhibitors, (b) a policy of active recruitment of organs together with surgical innovations (split liver, domino LT, living donor transplantation) contributing to the expansion of the pool of organs, (c) standardization of organ allocation policies, based on the sickest first policy. The applicability of LT, however, remains limited by the shortage of organs and the occurrence of long-term complications of immunosuppression. Due to the lack of effective alternative perspectives to LT for the treatment of end-stage liver diseases, the two major challenges for the liver transplant community should be the optimization of organ recruitment and the development of innovative immunosuppressive regimens able to overcome the side effects of current immunosuppressive drugs. The development of non heart beating donation and appropriate use of intrafamilial donation could partly compensate for the organ shortage in the midterm. The identification of molecular signatures in tolerant patients in whom immunosuppression could be stopped, and induction of tolerance, trough lymphocyte depletion or T lymphocyte costimulation blockade, are the most advanced research ways to reduce complications of immunosuppression. PMID- 19556085 TI - [Colorectal cancer discovered during pregnancy]. AB - Colorectal carcinoma during pregnancy is very rare. Prognosis is severe because of the advanced stage due to delay in diagnosis and the aggressiveness of this widespread metastatic disease. We present two cases of colorectal cancers, discovered during pregnancy, as well as the results of a literature review to apprehend this clinical situation, and we describe recommended medical and surgical management. PMID- 19556086 TI - [Cholestasis and cholestatic liver diseases]. AB - The main determinant of bile formation is an osmotic filtration process resulting from active transport of bile acids and other osmotic solutes (glutathion). Most of the membrane transporters ensuring bile formation have now been identified. The expression of these membrane transporters is regulated through transcriptional and post-traductional mechanisms. Transcriptional regulation is under the control of nuclear receptors activated by ligands such as bile acids, which act as endogenous steroids synthesized from cholesterol in hepatocytes. Cholestatic liver diseases comprise genetic diseases resulting from the complex interaction between genetic and environmental factors. Monogenic cholestatic diseases recently identified illustrate the key role of membrane transporters in biliary function. Bile acids and inflammatory mediators are potent modulators of transporters and nuclear receptor genes and thus trigger an adaptative response to cholestasis. The extent of this adaptative response could explain the compelling phenotypic variability of cholestatic diseases in childhood and adults. The first-line medical treatment is currently ursodeoxycholic acid and in case of failure of this medical treatment, liver transplantation is required. Recent progress in the molecular pathogenesis of bile formation and cholestatic liver diseases is expected to provide the design of drugs targeted to the molecular abnormalities typical of cholestatic diseases. PMID- 19556087 TI - Diagnosis of arterial sequestration using multidetector CT angiography. AB - BACKGROUND: Arterial sequestration is a rare congenital disorder. The diagnostic evaluation of this condition using multidetector computerized tomography (MDCT) has not been described previously. The purpose of this study was to describe the characteristic features of this disorder and to assess the use of MDCT in visualizing the characteristic anatomic features. METHODS: We retrospectively reviewed the records of seven patients with anomalous systemic blood supply to left lower lobe of the lung. All the patients had undergone MDCT angiography. To evaluate the lung parenchyma, bronchial branching pattern, and vascular anatomy, four series of images were systematically reconstructed as follows: contiguous transverse CT scans viewed at mediastinal and pulmonary window settings, oblique coronal and sagittal maximum intensity projections (MIP), multiplanar reconstruction (MPR), and three-dimensional volume-rendered images (VR) of airway and thoracic vascular structures. RESULTS: All 7 cases had isolated and tortuous arterial anatomy from the descending thoracic aorta to the basal segment of the left lower lobe; however, variable distribution of branches was observed. Characteristic findings of anomalous systemic arterial (ASA) supply were distinct from those seen in other pulmonary sequestration syndromes and were well visualized by the use of noninvasive MDCT. CONCLUSION: Complex CT findings allow clear imaging of arterial sequestration and the ASA blood supply; MDCT angiography has demonstrated its value and accuracy in diagnosing this condition, obviating the use of digital subtraction angiography and magnetic resonance angiography for the diagnosis of arterial sequestration. PMID- 19556088 TI - Non-contrast-enhanced hepatic MR angiography: do two-dimensional parallel imaging and short tau inversion recovery methods shorten acquisition time without image quality deterioration? AB - OBJECTIVE: To study whether shortening the acquisition time for selective hepatic artery visualization is feasible without image quality deterioration by adopting two-dimensional (2D) parallel imaging (PI) and short tau inversion recovery (STIR) methods. MATERIALS AND METHODS: Twenty-four healthy volunteers were enrolled. 3D true steady-state free-precession imaging with a time spatial labeling inversion pulse was conducted using 1D or 2D-PI and fat suppression by chemical shift selective (CHESS) or STIR methods. Three groups of different scan conditions were assigned and compared: group A (1D-PI factor 2 and CHESS), group B (2D-PI factor 2*2 and CHESS), and group C (2D-PI factor 2*2 and STIR). The artery-to-liver contrast was quantified, and the quality of artery visualization and overall image quality were scored. RESULTS: The mean scan time was 9.5+/-1.0 min (mean+/-standard deviation), 5.9+/-0.8 min, and 5.8+/-0.5 min in groups A, B, and C, respectively, and was significantly shorter in groups B and C than in group A (P<0.01). The artery-to-liver contrast was significantly better in group C than in groups A and B (P<0.01). The scores for artery visualization and overall image quality were worse in group B than in groups A and C. The differences were statistically significant (P<0.05) regarding the arterial branches of segments 4 and 8. Between group A and group C, which had similar scores, there were no statistically significant differences. CONCLUSION: Shortening the acquisition time for selective hepatic artery visualization was feasible without deterioration of the image quality by the combination of 2D-PI and STIR methods. It will facilitate using non-contrast-enhanced MRA in clinical practice. PMID- 19556089 TI - A better understanding of urogenital tuberculosis pathophysiology based on radiological findings. AB - PURPOSE: To assess the radiological findings of urogenital tuberculosis (UGT) in patients at different disease stages, for a better understanding of its pathophysiology. PATIENTS AND METHODS: We retrospectively reviewed the radiological exams of 20 men (median age 41 years; range: 28-65) with urogenital tuberculosis diagnosis. The patients were classified in the following groups: (1) bilateral renal tuberculosis with predominantly parenchymatous involvement; (2) unilateral renal tuberculosis; (3) unilateral renal tuberculosis with bladder tuberculosis and (4) bilateral renal tuberculosis with bladder tuberculosis. RESULTS: One AIDS patient had multiple bilateral renal tuberculosis abscesses (group 1). Six patients had unilateral renal tuberculosis with hydronephrosis due to stenosis and thickening of the collecting system, without involvement of the bladder or contralateral kidney (group 2). Six patients had bladder tuberculosis with diffuse thickening of the bladder wall, with one very low or no function kidney while the other kidney was normal (group 3). Seven patients had bladder tuberculosis associated to a very low or no function kidney with the other kidney with high-grade vesicoureteral reflux-associated ureterohydronephrosis (group 4). In two patients, sequential exams showed evolution of tuberculosis from a unilateral renal and ureteral lesion to contracted bladder and dilatation of the contralateral kidney secondary to high-grade reflux. CONCLUSIONS: UGT may have variable radiological presentations. However, in two of our cases we have seen that tuberculosis involvement of the urinary tract may be sequential. Further evidences are necessary to confirm this hypothesis. PMID- 19556090 TI - Multilocus sequence analysis of bradyrhizobia isolated from Aeschynomene species in Senegal. AB - This study reports the multilocus sequence analysis (MLSA) of nine house-keeping gene fragments (atpD, dnaK, glnA, glnB, gltA, gyrB, recA, rpoB and thrC) on a collection of 38 Bradyrhizobium isolated from Aeschynomene species in Senegal, which had previously been characterised by several phenotypic and genotypic techniques, allowing a comparative analysis of MLSA resolution power for species delineation in this genus. The nifH locus was also studied to compare house keeping and symbiotic gene phylogenies and obtain insights into the unusual symbiotic properties of these Aeschynomene symbionts. Phylogenetic analyses (maximum likelihood, Bayesian) of concatenated nine loci produced a well-resolved phylogeny of the strain collection separating photosynthetic bradyrhizobial strains (PB) from non-photosynthetic bradyrhizobial (NPB) ones. The PB clade was interpreted as the remains an expanding ancient species that presently shows high diversification, giving rise to potential new species. B. denitrificans LMG8443 and BTAi1 strains formed a sub-clade that was identified as recently emerging new species. Congruence analyses (by Shimodaira-Hasegawa (S-H) tests) identified three gene-fragments (dnaK, glnB and recA) that should be preferred for MLSA analyses in Bradyrhizobium genus. The nine loci or nifH phylogenies were not correlated with the unusual symbiotic properties of PB (nod-dependent/nod independent). Advantages and drawbacks of MLSA for species delineation in Bradyrhizobium are discussed. PMID- 19556091 TI - Semi-preparative isolation and structural elucidation of an impurity in citalopram by LC/MS/MS. AB - Two impurities were detected in citalopram bulk drug substance by HPLC analysis. A new LC-ESI/MS method was developed for the identification of impurities. One of the impurities was found to be unknown and has not been reported previously. The structure of the unknown impurity was proposed on the basis of MS(n) data obtained using ion trap mass analyzer and accurate mass obtained using Q-TOF mass analyzer. The impurity was isolated by semi-preparative HPLC. The structure of the impurity was confirmed as 1-(1,1-bis (4-fluorophenyl)-1,3 dihydroisobenzofuran-5-yl)-4-(dimethylamino) butan-1-one hydrobromide by using NMR and IR spectroscopy. PMID- 19556092 TI - Validation of an LC-MS/MS assay of terpene trilactones in Ginkgo biloba extracts and pharmaceutical formulations through standard addition method. AB - A chromatographic column packed with 1.8 microm particle size octadecyl modified silicagel was used to separate terpene trilactones from Ginkgo biloba extracts/pharmaceutical formulations. Gradient elution was applied, using acidic methanol and water as mobile phase components (0.1% formic acid addition). No specific sample preparation is needed, except dissolution/extraction in methanol of the solid material. Baseline separation of bilobalide and ginkgolides A, B, C and J is obtained within 4 min. The gradient profile is needed to elute the remaining matrix from column. A separation cycle takes 7 min, including column re equilibration. MS/MS detection with positive electrospray ionization and triple quadrupole mass analysis was used. Multiple reaction monitoring mode was applied for data acquisition, taking protonated molecular ions as precursors. In order to generate reproducible ionization conditions, the standard addition method was considered. The assay of terpene trilactones obtained under such conditions was validated according to guidances in place. Method intermediate reproducibility corresponds to a relative standard deviation of less than 10%. Accuracy, expressed in terms of absolute percent bias ranged from 90% to 110%. Spectral confirmation of target analytes was also included in the validation procedure. PMID- 19556093 TI - Development of biosensor-based SPR technology for biological quantification and quality control of pharmaceutical proteins. AB - We developed and validated a biosensor-based surface plasmon resonance (SPR) technology for the biological quantification and quality control (QC) of pharmaceutical proteins using reference materials as the standard. The surface of the receptors was made homogeneous by covalently immobilizing the receptors onto Au-membrane microchips for use as biosensors for reliably detecting the activity of drug proteins. This assay used only limited amounts of ligands and no additional detection agents. The products were determined to have binding capacity equivalent to that of the reference materials and to exhibit a recovery range of 88.4-115.0%. The binding of analytes to the specific ligand is concentration dependent and parallel. CTLA-4 fusion proteins were quantitatively detectable at concentrations as low as 125 ng/mL. The intra-assay precision was in the range of 1.07-7.27%, and the inter-lot precision was 13.03%. These data proved the usefulness of improved biosensor-based assays in biological quantification and QC of pharmaceutical proteins. This approach is an alternative to traditional assays and offers a potentially significant advantage in that the microchip can be regenerated thus enabling multiple analyses to be performed with a single sensor. PMID- 19556094 TI - Interest in marijuana treatment programs among teenage smokers and nonsmokers. AB - Little is known about adolescents' interest in marijuana treatment programs. This question was evaluated by telephone interview in a convenience sample of 575 adolescents responding to advertisements for tobacco research studies. Eighty-one percent of respondents endorsed the need for marijuana treatment programs for adolescents. These adolescents were younger and less likely to smoke tobacco, smoke marijuana, or use alcohol than those not endorsing such a need. Among the 192 marijuana smokers, the 58.8% who endorsed the need for marijuana treatment programs took their first puff of marijuana at a younger age than those who did not endorse the need. Those who were willing to participate in a marijuana treatment program were more likely African American and took their first marijuana puff at a younger age than those not interested in treatment. These findings suggest that most adolescent marijuana smokers endorse the need for and are willing to attend marijuana treatment programs. PMID- 19556095 TI - Impact of Access to Recovery services on alcohol/drug treatment outcomes. AB - The purpose of this study was to assess the impact of providing recovery support services to clients receiving publicly funded chemical dependency (CD) treatment through the Access to Recovery (ATR) Program in Washington State. Services included case management, transportation, housing, and medical. A comparison group composed of clients who received CD treatment only was constructed using a multistep procedure based on propensity scores and exact matching on specific variables. Outcomes were obtained from administrative data sources. Results indicated that ATR services were associated with a number of positive outcomes including increased length of stay in treatment, increased likelihood of completing treatment, and increased likelihood of becoming employed. The beneficial effects of ATR services on treatment retention were most pronounced when they were provided between 31 and 180 days after treatment began. The results reported here offer evidence for the value of ATR services. PMID- 19556096 TI - The pre-operative stratification of patients with colorectal liver metastases: computed tomography arterial portography (CTAP) has no added value. AB - AIMS: The purpose of this study was to determine if Computed Tomography Arterial Portography (CTAP) has additional value to Contrast Enhanced helical CT (CE-CT) in selecting patients for hepatic surgery or Isolated Hepatic Perfusion/systemic chemotherapy. MATERIALS AND METHODS: Forty-one patients were included. All CT's were performed in the normal pre-operative work-up of patients with liver metastases in our regular clinical setting and reviewed blinded by a radiologist. For CE-CT and CTAP the number, size (largest diameter) and location of all suspected malignant liver lesions were recorded. The favourable treatment option was determined based on the results of CE-CT and CTAP independently. The therapeutic decision based on CE-CT and CTAP was compared with the definite treatment. For all patients with recorded findings during surgery, consisting of intra-operative ultrasound, liver palpation and histology a standard of reference for lesion detection was available. For these patients detection rates and the fraction of false positive lesions were calculated. RESULTS: Twenty-seven patients were treated with hepatic resection and/or RFA. Fourteen patients were treated with chemotherapy, 4 with Isolated Hepatic Perfusion (IHP) and 10 with systemic therapy. Based on the findings on CE-CT 31 patients were classified as surgical candidates and 10 as non-surgical patients. Based on the findings on CTAP, surgery should be the treatment of choice in 29 patients and 12 patients were classified non-surgical. CE-CT and CTAP disagreed in two cases (4.9%). Seventy-four metastases were identified at surgery and pathologically proved. CE CT and CTAP showed 53 (.72) and 66 (.89) metastases, respectively. CONCLUSION: Despite a significantly higher detection rate for hepatic metastases, CTAP has no added value in the therapeutic stratification in candidates for resection of hepatic metastases of colorectal cancer. PMID- 19556097 TI - Cementless femoral components in young patients: review and meta-analysis of total hip arthroplasty and hip resurfacing. AB - The study purpose was to analyze current results of modern cementless femoral components in young patients having total hip arthroplasty (THA) or hip resurfacing. Twenty-two studies (n = 5907; hips = 6408) evaluating modern cementless THA in young patients and 15 studies evaluating hip resurfacing (n = 3002; hips = 3269) were included. Meta-analysis techniques were used to pool failure rates. The pooled failure rate for THA using femoral revision for mechanical failure as an end point was 1.3% (95% confidence interval [CI], 1.0% 1.7%) at a mean 8.4 years of follow-up. At a mean of 3.9 years of follow-up, the pooled mechanical failure rate of the femoral component for hip resurfacing was 2.6% (95% CI, 2.0-3.4). In conclusion, the enthusiasm for hip resurfacing should be tempered by these data. Longer follow-up and direct comparison trials are required to confirm these findings. PMID- 19556098 TI - A self-report measure of subtle avoidance and safety behaviors relevant to social anxiety: development and psychometric properties. AB - According to cognitive theories, safety-seeking behaviors are crucial in both the maintenance and management of social anxiety. In order to facilitate assessment of these behaviors the Subtle Avoidance Frequency Examination (SAFE) was developed. Three factors emerged from the SAFE, which appeared to reflect active "safety" behaviors, subtle restriction of behavior, and behaviors aimed at avoiding or concealing physical symptoms. The SAFE demonstrated strong internal consistency, good construct validity and the ability to discriminate between clinical and non-clinical participants. In addition, the SAFE was responsive to the effects of treatment. Given its excellent psychometric properties, the SAFE may be useful to further investigate the role of safety strategies in social anxiety and to assess treatment outcomes. PMID- 19556099 TI - Social phobia and perceived friendship quality. AB - Although it is clear that people with social phobia have interpersonal impairment, evidence that social phobia (as opposed to other mental disorders) affects friendship in particular is lacking. Two large epidemiological datasets were used to test whether diagnosis of social phobia is related to perceived friendship quality above and beyond perceived family relationship quality, diagnosis of other mental disorders, and a variety of demographic variables. After Bonferroni correction, social phobia was the only diagnosis related to perceived friendship quality above and beyond other factors, such that people with social phobia reported more impaired friendship quality. Social phobia's effect was similar in magnitude to demographic characteristics in both samples. The current study demonstrates that social phobia is specifically related to perceived friendship quality, suggesting that this aspect of social phobia's effects is worthy of further study. PMID- 19556100 TI - Acute debranching and stent grafting for a ruptured penetrating ulcer of the aortic arch. AB - BACKGROUND: We report a hybrid treatment for a ruptured penetrating aortic ulcer (PAU) of the aortic arch in emergency conditions. METHODS: A 74-year-old man with severe comorbidity was diagnosed with a symptomatic rupture of a PAU of the aortic arch. As he was considered unfit for conventional open repair and it was an emergency condition, an acute endovascular repair with cervical debranching of the supra-aortic trunks could be proposed. The left common carotid artery and left subclavian artery were first debranched and revascularized through cervical access, and the endovascular stent graft was then deployed via femoral access in the aortic arch. At >or=2 years the patient is safe with no sign of progression of the disease. CONCLUSION: Acute debranching and stent grafting is feasible in emergency conditions to treat ruptured PAU of the aortic arch, with satisfying short- and mid-term outcomes, and can be proposed to patients considered unfit for conventional open repair. PMID- 19556101 TI - Utility of intravascular ultrasound examination during carotid stenting. AB - BACKGROUND: Intravascular ultrasound (IVUS) examination may provide useful information during endovascular procedures. However, its actual clinical utility for carotid stenting remains unclear. We evaluated the usefulness of IVUS as a complementary tool during endovascular procedures in the carotid arteries and its impact on the modification of the therapeutic strategy. METHODS: Between April 2006 and April 2007, 18 symptomatic patients (nine with transient ischemic attack, nine with stroke) underwent angioplasty and stenting of >70% stenosis of the internal carotid artery (ICA). Target lesions were identified with preoperative duplex scanning and further confirmed at angiography. Intraoperative IVUS examination was performed in all patients. Multilevel measurement of the artery as well as virtual histology images, before and after stenting, were obtained. RESULTS: The technique was 100% successful. There were no complications at the femoral puncture site and no neurological or other events. Compared with angiography, IVUS findings showed an average arterial diameter that was greater than 1.64+/-0.22mm. Based on virtual histology information, endovascular treatment was excluded in two patients and carotid endarterectomy was performed. Type or size of the stent was modified after IVUS examination in eight cases. CONCLUSION: IVUS examination provides complementary information to that obtained using conventional diagnostic procedures. It may be useful for characterizing the composition and measurement of the target lesion, choosing the type and size of the stent, and evaluating the results after the procedure. PMID- 19556102 TI - An initial assessment of the psychometric properties of the Complicated Grief Questionnaire for People with Intellectual Disabilities (CGQ-ID). AB - Given the research evidence that people with intellectual disability (ID) do grieve following bereavement, the present study aimed to describe and gather preliminary psychometric data for a version of the Inventory of Complicated Grief [Prigerson, H. G., Maciejewski, P. K., Reynolds, C. F., Bierhals, A. J., Newsom, J. T., Fasiczka, A., et al. (1995). Inventory of Complicated Grief: A scale to measure maladaptive symptoms of loss. Psychiatry Research, 59, 65-79] adapted for use with this population. Carers completed the Complicated Grief Questionnaire for People with ID (CGQ-ID) for 76 individuals with ID, half of whom had experienced a parental bereavement within the last 2 years. The final scale and subscales (Separation Distress and Traumatic Grief) showed very good internal and inter-rater reliability and distinguished between the two groups. While the findings suggest that the CGQ-ID is suitable for identifying complicated grief type symptoms among adults with ID, further research must be conducted to ascertain whether the findings can be replicated. PMID- 19556103 TI - Learning a multi-dimensional companding function for lossy source coding. AB - Although the importance of lossy source coding has been growing, the general and practical methodology for its design has not been completely resolved. The well known vector quantization (VQ) can represent any fixed-length lossy source coding, but requires too much computation resource. Companding vector quantization (CVQ) can reduce the complexity of non-structured VQ by replacing vector quantization with a set of scalar quantizations and can represent a wide class of practically useful VQs. Although an analytical derivation of optimal CVQ is difficult except for very limited cases, optimization using data samples can be performed instead. Here we propose a CVQ optimization method, which includes bit allocation by a newly derived distortion formula as a generalization of Bennett's formula, and test its validity. We applied the method to transform coding and compared the performance of our CVQ with those of Karhunen-Loeve transformation (KLT)-based coding and non-structured VQ. As a consequence, we found that our trained CVQ outperforms not only KLT-based coding but also non structured VQ in the case of high bit-rate coding of linear mixtures of uniform sources. We also found that trained CVQ even outperformed KLT-based coding in the low bit-rate coding of a Gaussian source. To highlight the advantages of our approach, we also discuss the degradation of non-structured VQ and the limitations of theoretical analyses which are valid for high bit-rate coding. PMID- 19556104 TI - An online incremental learning pattern-based reasoning system. AB - An architecture for reasoning with pattern-based if-then rules is proposed. By processing patterns as real-valued vectors and classifying similar if-then rules into clusters in long-term memory, the proposed system can store pattern-based if then rules of propositional logic, including conjunctions, disjunctions, and negations. Moreover, it achieves some important properties for intelligent systems such as incremental learning, generalization, avoidance of duplicate results, and robustness to noise. Results of experiments demonstrate that the proposed method is effective for intelligent systems for solving various tasks autonomously in a real environment. PMID- 19556105 TI - Evolutionary artificial neural networks by multi-dimensional particle swarm optimization. AB - In this paper, we propose a novel technique for the automatic design of Artificial Neural Networks (ANNs) by evolving to the optimal network configuration(s) within an architecture space. It is entirely based on a multi dimensional Particle Swarm Optimization (MD PSO) technique, which re-forms the native structure of swarm particles in such a way that they can make inter dimensional passes with a dedicated dimensional PSO process. Therefore, in a multidimensional search space where the optimum dimension is unknown, swarm particles can seek both positional and dimensional optima. This eventually removes the necessity of setting a fixed dimension a priori, which is a common drawback for the family of swarm optimizers. With the proper encoding of the network configurations and parameters into particles, MD PSO can then seek the positional optimum in the error space and the dimensional optimum in the architecture space. The optimum dimension converged at the end of a MD PSO process corresponds to a unique ANN configuration where the network parameters (connections, weights and biases) can then be resolved from the positional optimum reached on that dimension. In addition to this, the proposed technique generates a ranked list of network configurations, from the best to the worst. This is indeed a crucial piece of information, indicating what potential configurations can be alternatives to the best one, and which configurations should not be used at all for a particular problem. In this study, the architecture space is defined over feed-forward, fully-connected ANNs so as to use the conventional techniques such as back-propagation and some other evolutionary methods in this field. The proposed technique is applied over the most challenging synthetic problems to test its optimality on evolving networks and over the benchmark problems to test its generalization capability as well as to make comparative evaluations with the several competing techniques. The experimental results show that the MD PSO evolves to optimum or near-optimum networks in general and has a superior generalization capability. Furthermore, the MD PSO naturally favors a low-dimension solution when it exhibits a competitive performance with a high dimension counterpart and such a native tendency eventually yields the evolution process to the compact network configurations in the architecture space rather than the complex ones, as long as the optimality prevails. PMID- 19556106 TI - Comprehensive evaluation of left ventricular strain using speckle tracking echocardiography in normal adults: comparison of three-dimensional and two dimensional approaches. AB - OBJECTIVE: The two-dimensional speckle tracking (2DT) method is based on the measurements of strain on two-dimensional (2D) images, ignoring actual three dimensional (3D) myocardial movements. We sought to investigate the feasibility of the newly developed three-dimensional speckle tracking (3DT) method to assess longitudinal, circumferential, and radial strain values, and then compared the data with those measured by 2DT. METHODS: Echocardiographic examinations were performed in 46 volunteers. In the apical 3D volumetric images, 3 vectors of the strains were analyzed in 16 myocardial segments. 2D longitudinal strain was assessed in apical 4-, 3-, and 2-chamber views, and circumferential and radial strains were measured in parasternal short-axis view. RESULTS: The average time for 3D image acquisition and 3D strain analysis by 3DT was significantly shorter than for 2DT. Longitudinal strain value by 3DT was significantly smaller than by 2DT (-17.4% +/- 5.0% vs -19.9% +/- 6.7%, P < .0001), and circumferential strain value by 3DT was significantly larger than by 2DT (-30.1% +/- 7.1% vs -26.3% +/- 6.9%, P < .0001). Intraobserver and interobserver variabilities were 10.1% and 10.9% in 3DT, and 9.9% and 11.1% in 2DT, respectively. CONCLUSION: 3DT is a simple, feasible, and reproducible method to measure longitudinal, circumferential, and radial strains. The discordant results between 3DT and 2DT may be explained by the 3D cardiac motion that has been ignored in current 2DT. PMID- 19556107 TI - Xp22.3 microdeletions in three Chinese families with X-linked ichthyosis. PMID- 19556108 TI - Autosomal recessive congenital ichthyosis and congenital hypothyroidism in a Tunisian patient with a nonsense mutation in TGM1. PMID- 19556109 TI - ECT rekindles pharmacological response in schizophrenia. AB - OBJECTIVE: The aim of our naturalistic-observational study was to determine the efficacy and utility of electroconvulsive therapy (ECT) in clinical population of individuals with schizophrenia where pharmacological response was suboptimal. METHODS: The cohort comprised 27 patients suffering from schizophrenia with refractoriness to antipsychotic agents and with severe, disabling symptoms. They only interventional assessing tool was clinical global impression (CGI-S) performed at the baseline and at the end of the treatment. RESULTS: The administration of ECT resulted in overall clinical improvement reflected in CGI scales and descriptions in clinical notes. On 12 months follow-up period, 10 patients (37.1%) maintained improvement and were able to continue with pharmacological therapy only, suggesting its rekindling effect, especially with Clozapine. Conversely, 17 patients (62.9%) deteriorated and required an additional course of ECT to maintain improvement. In some cases, maintenance ECT treatment was required. The group who engaged in self-harming behaviour at baseline demonstrated they were more likely to relapse into psychosis at the end of the first course of ECT, their self-harming behaviour abated, especially when maintenance ECT was undertaken. CONCLUSIONS: Although limited by lack of control group, small sample size, heterogeneous symptom profiles and various concurrent neuroleptic agents, the ECT proved as valuable and safe augmentative procedure when unsatisfactory response to pharmacological interventions had been demonstrated prior to interventions. This effect was evident despite the chronicity of the illness. PMID- 19556110 TI - Prospects for the classification of mental disorders in women. AB - Many mental disorders show marked gender differences as regards prevalence, symptomatology, risk factors or course. Other disorders do per definition only occur in women - e.g. premenstrual dysphoric disorder (PMDD) - or are markedly influenced by female specific factors such as hormonal changes over the life cycle or during reproductive processes. Current classification systems have tried to take into account these gender aspects, but some problems will certainly have to be discussed again with the next revisions of the ICD and DSM. As regards gender differences in prevalence and symptomatology questions of gender bias in diagnostic instruments and diagnostic criteria will have to be readdressed. New findings from unselected epidemiological samples, which were analysed by gender, will have to be taken into account as well as new findings from research into gender specific personality traits, which can influence the symptomatology of mental disorders. Decisions will have to be taken whether to revise existing diagnostic criteria and provide alternative diagnostic thresholds for men and women or even to develop alternative criteria sets in certain disorders, or rather to enhance the gender neutrality of criteria. A further question to be addressed will be that of gender specific diagnoses versus diagnostic specifiers. In the whole discussion two main aims of classification should be given priority: the research aim of identifying genuine entities with a common aetiology, which means we should be able to identify specific diagnostic entities with descriptive, construct, and predictive validity quite independently of the influences of gender; and the clinical aim to improve treatment and care for men and women, which often means to offer gender-specific approaches. PMID- 19556111 TI - Attached to the web--harmful use of the Internet and its correlates. AB - BACKGROUND AND AIMS: The aim of this study was to test the validity of the Finnish version of the Internet Addiction Test and the correlates of harmful use of the Internet. METHODS: One thousand eight hundred and twenty-five students (45.5% men and 54.5% women, mean age 24.7 years, S.D.=5.7) filled in a web-based questionnaire including IAT, reasons for use of the Internet, distress, social support, and substance use. RESULTS: Men had a statistically significantly higher mean score on the IAT than women. Subjects with self-reported use of cannabis had higher mean score on the IAT compared to non-users (39.5 [11.3] vs 35.8 [10.8]). The total IAT score was associated with "adult entertainment" (OR=1.07, 95%CI: 1.06-1.08, P<0.001), "playing games" (OR=1.05, 95%CI: 1.04-1.06, P<0.001), "chatting" (OR=1.07, 95%CI: 1.06-1.08, P<0.001) and "discussion" (OR=1.08, 95%CI: 1.07-1.09, P<0.001) as reasons for Internet use. The IAT score had a significant negative correlation with social support (r=-0.24, P<0.001) and a significant positive correlation with the CAGE score (r=0.18, P<0.001). Using factor analysis, we found a single factor solution with a Cronbach's alpha of 0.92. CONCLUSIONS: The IAT seems to provide a valid measurement of harmful use of the Internet, as the score was significantly associated with variables tapping psychopathology. PMID- 19556113 TI - Non-specific adhesion on biomaterial surfaces driven by small amounts of protein adsorption. AB - This work explores how long-range non-specific interactions, resulting from small amounts of adsorbed fibrinogen, potentially influence bioadhesion. Such non specific interactions between protein adsorbed on a biomaterial and approaching cells or bacteria may complement or even dominate ligand-receptor mating. This work considers situations where the biomaterial surface and the approaching model cells (micron-scale silica particles) exhibit strong electrostatic repulsion, as may be the case in diagnostics and lab-on-chip applications. We report that adsorbed fibrinogen levels near 0.5mg/m(2) produce non-specific fouling. For underlying surfaces that are less fundamentally repulsive, smaller amounts of adsorbed fibrinogen would have a similar effect. Additionally, it was observed that particle adhesion engages sharply and only above a threshold loading of fibrinogen on the collector. Also, in the range of ionic strength, I, below about 0.05M, increases in I reduce the fibrinogen needed for microparticle capture, due to screening of electrostatic repulsions. Surprisingly, however, ionic strengths of 0.15M reduce fibrinogen adsorption altogether. This observation opposes expectations based on DLVO arguments, pointing to localized electrostatic attractions and hydration effects to drive silica-fibrinogen adhesion. These behaviors are benchmarked against microparticle binding on silica surfaces carrying small amounts of a polycation, to provide insight into the role of electrostatics in fibrinogen-driven non-specific adhesion. PMID- 19556114 TI - Isotherm, kinetic and thermodynamic studies of lead and copper uptake by H2SO4 modified chitosan. AB - The kinetic and thermodynamic adsorption and adsorption isotherms of Pb(II) and Cu(II) ions onto H(2)SO(4) modified chitosan were studied in a batch adsorption system. The experimental results were fitted using Freundlich, Langmuir and Dubinin-Radushkevich isotherms; the Langmuir isotherm showed the best conformity to the equilibrium data. The pseudo-first order, pseudo-second order and intraparticle diffusion kinetic models were employed to analyze the kinetic data. The adsorption behavior of Pb(II) and Cu(II) was best described by the pseudo second order model. Thermodynamic parameters such as free energy change (DeltaG degrees ), enthalpy change (DeltaH degrees ) and entropy change (DeltaS degrees ) were determined; the adsorption process was found to be both spontaneous and exothermic. No physical damage to the adsorbents was observed after three cycles of adsorption/desorption using EDTA and HCl as eluents. The mechanistic pathway of the Pb(II) and Cu(II) uptake was examined by means of Fourier transform infrared (FTIR) and Energy dispersive X-ray (EDX) spectroscopy. The equilibrium parameter (R(L)) indicated that chitosan-H(2)SO(4) was favorable for Pb(II) and Cu(II) adsorption. PMID- 19556112 TI - Oral glycine administration increases brain glycine/creatine ratios in men: a proton magnetic resonance spectroscopy study. AB - Oral high-dose glycine administration has been used as an adjuvant treatment for schizophrenia to enhance glutamate neurotransmission and mitigate glutamate system hypofunction thought to contribute to the disorder. Prior studies in schizophrenia subjects documented clinical improvements after 2 weeks of oral glycine administration, suggesting that brain glycine levels are sufficiently elevated to evoke a clinical response within that time frame. However, no human study has reported on brain glycine changes induced by its administration. We utilized a noninvasive proton magnetic resonance spectroscopy ((1)H-MRS) technique termed echo time-averaged (TEAV) (1)H-MRS, which permits noninvasive quantification of brain glycine in vivo, to determine whether 2 weeks of oral glycine administration (peak dose of 0.8 g/kg/day) increased brain glycine/creatine (Gly/Cr) ratios in 11 healthy adult men. In scans obtained 17 h after the last glycine dose, brain (Gly/Cr) ratios were significantly increased. The data indicate that it is possible to measure brain glycine changes with proton spectroscopy. Developing a more comprehensive understanding of human brain glycine dynamics may lead to optimized use of glycine site agonists and glycine transporter inhibitors to treat schizophrenia, and possibly to treat other disorders associated with glutamate system dysfunction. PMID- 19556115 TI - Benzylpenicillin, acetylcysteine and silibinin as antidotes in human hepatocytes intoxicated with alpha-amanitin. AB - Fatalities due to mushroom poisonings are increasing worldwide, with high mortality rate resulting from ingestion of amanitin-producing species. Intoxications caused by amanitin-containing mushrooms represent an unresolved problem in clinical toxicology since no specific and fully efficient antidote is available. The objective of this study was a comparative evaluation of benzylpenicillin (BPCN), acetylcysteine (ACC) and silibinin (SIL) as an antidotes in human hepatocytes intoxicated with alpha-amanitin (alpha-AMA). All experiments were performed on cultured human hepatocytes. Cytotoxicity evaluation of cultured cells using MTT assay and measurement of lactate dehydrogenase (LDH) activity was performed at 12, 24 and 48h of exposure to alpha-AMA and/or antidotes. The significant decline of cell viability and significant increase of LDH activity were observed in all experimental hepatocyte cultures after 12, 24 and 36h exposure to alpha-AMA at concentration 2microM. Exposure of the cells to alpha AMA resulted also in significant reduction of cell spreading and attachment. However, addition of tested antidotes to experimental cultures significantly stimulated cell proliferation and attachment. In cell cultures exposed simultaneously to alpha-AMA and tested antidotes cytotoxic parameters (MTT and LDH) were not significantly different from control incidences. The cytoprotective effect of all antidotes was not dose-related, which reflects a high efficacy of all these substances. Administration of studied antidotes was not associated with any adverse effects in hepatocytes. The administration of ACC, BPCN or SIL to human hepatocyte cultures showed a similar strong protective effect against cell damage in alpha-AMA toxicity. PMID- 19556116 TI - Tiotropium and risk for fatal and nonfatal cardiovascular events in patients with chronic obstructive pulmonary disease: systematic review with meta-analysis. AB - BACKGROUND: There are safety concerns regarding the use of anticholinergics in the COPD patient population. The purpose of this review was to evaluate the cardiovascular risk of regular use of inhaled tiotropium bromide in patients with COPD of any severity. METHODS: Systematic searches were conducted in MEDLINE, EMBASE, the Cochrane Controlled Trials Register, manufactures' trial register, and FDA databases, without language restriction. Primary outcomes were a composite of major adverse cardiovascular events, cardiovascular mortality, and nonfatal myocardial infarction (MI) or stroke during the treatment period. Relative risks (RR) were estimated using fixed-effects models and statistical heterogeneity was estimated with the I2 statistic. RESULTS: Nineteen randomized controlled trials (18,111 participants) were selected. There was no difference in the incidence of adverse cardiovascular events (RR=0.96; 95% CI, 0.82-1.12, I2=6%). Among individual components of the composite outcome, tiotropium did not significantly increase the risk of cardiovascular death (RR=0.93; 95% CI, 0.73 1.20, I2=1%), nonfatal MI (RR=0.84; 95% CI, 0.64-1.09, I2=0%), and nonfatal stroke (RR=1.04; 95% CI, 0.78-1.39, I2=0%). A smoking history of > or = 55 pack years presented a trend to a higher rate of cardiovascular adverse events in patients receiving tiotropium. CONCLUSIONS: Compared with control (placebo or salmeterol), tiotropium did not significantly increase the risk of adverse major cardiovascular events among COPD patients. Subgroup analysis suggested that smoking history can modify the risk of cardiovascular adverse events. PMID- 19556117 TI - Intermunicipal cooperation, privatization and waste management costs: evidence from rural municipalities. AB - The aim of this paper is to analyze the effects of intermunicipal cooperation and privatization on the delivery costs of urban solid waste services in rural environments. The results of our empirical analysis, which we conducted among a sample of very small municipalities, indicate that small towns that cooperate incur lower costs for their waste collection service. Cooperation also raises collection frequency and improves the quality of the service in small towns. By contrast, the form of production, whether it is public or private, does not result in systematic differences in costs. Interestingly, the degree of population dispersion, that is, the number of population units within the municipal jurisdiction, has a significant positive relation with service costs. No evidence of scale economies is found because small municipalities have likely exploited them by means of intermunicipal cooperation. PMID- 19556118 TI - Taste receptor cell-based biosensor for taste specific recognition based on temporal firing. AB - Taste receptor cells are the taste sensation elements expressing sour, salty, sweet, bitter and umami receptors, respectively. There are cell-to-cell communications between different types of cells. Nevertheless, the mechanism of taste sensation and taste information coded by taste receptor cell is not well understood at present and it is a long-standing issue. In order to explore taste sensation and analyze taste-firing responses from another point of view, we present a promising biomimetic taste receptor cell-based biosensor. The temporal firing responses to different tastants are recorded. Meanwhile, we investigate the firing rate and temporal firing of taste receptor cells. The experimental results are consistent with that from patch clamp and molecular biology experiment. Firing rate is dependent on the concentration of stimulus. PCA analysis (principal component analysis) of the temporal firing responses shows that the responses from different types of taste receptor cells can be distinguished. Furthermore, exogenous ATP is applied to mimic the effects of transmitter ATP (adenosine triphosphate) released from type II cells onto type III cells. Both enhanced and inhibitory effects on spontaneous firing are observed. This novel biomimetic hybrid biosensor provides a potential solution to investigate the taste sensation and coding mechanisms in a non-invasive way. PMID- 19556119 TI - Fabrication and characterization of a multiwell array SERS chip with biological applications. AB - Uniform, large surface area substrates for surface-enhanced Raman spectroscopy (SERS) are fabricated by oblique angle deposition. The SERS-active substrates are patterned by a polymer-molding technique to provide a uniform array for high throughput biosensing and multiplexing. Using a conventional SERS-active molecule, 1,2-di(4-pyridyl)ethylene (BPE) >or=98%, we show that this device provides a uniform Raman signal enhancement from well to well with a detection limit of at least 10(-8)M of the BPE solution or 10(-18)mol of BPE. The SERS intensity is also demonstrated to vary logarithmically with the log of BPE concentration and the apparent sensitivity of the patterned substrate is compared to previous reports from our group on non-patterned substrates. Avian influenza is analyzed to demonstrate the utility of SERS multiwell patterned substrates for biosensing. The spectra acquired from patterned substrates show better reproducibility and less variation compared to the unpatterned substrates according to multivariate analysis. Our results highlight potential advantages of the patterned substrate. PMID- 19556120 TI - Membrane-less cloth cathode assembly (CCA) for scalable microbial fuel cells. AB - One of the main challenges for scaling up microbial fuel cell (MFC) technologies is developing low-cost cathode architectures that can generate high power output. This study developed a simple method to convert non-conductive material (canvas cloth) into an electrically conductive and catalytically active cloth cathode assembly (CCA) in one step. The membrane-less CCA was simply constructed by coating the cloth with conductive paint (nickel-based or graphite-based) and non precious metal catalyst (MnO(2)). Under the fed-batch mode, the tubular air chamber MFCs equipped with Ni-CCA and graphite-CCA generated the maximum power densities of 86.03 and 24.67 mW m(-2) (normalized to the projected cathode surface area), or 9.87 and 2.83 W m(-3) (normalized to the reactor liquid volume), respectively. The higher power output of Ni-CCA-MFC was associated with the lower volume resistivity of Ni-CCA (1.35 x 10(-2)Omega cm) than that of graphite-CCA (225 x 10(-2)Omega cm). At an external resistance of 100 Omega, Ni CCA-MFC and graphite-CCA-MFC removed approximately 95% COD in brewery wastewater within 13 and 18d, and achieved coulombic efficiencies of 30.2% and 19.5%, respectively. The accumulated net water loss through the cloth by electro-osmotic drag exhibited a linear correlation (R(2)=0.999) with produced coulombs. With a comparable power production, such CCAs only cost less than 5% of the previously reported membrane cathode assembly. The new cathode configuration here is a mechanically durable, economical system for MFC scalability. PMID- 19556122 TI - A phase I and pharmacokinetic study of gemcitabine given by 24-h hepatic arterial infusion. AB - PURPOSE: This study was performed to assess the toxicities, the maximum-tolerated dose (MTD), the pharmacokinetics and the anti-tumour activity of gemcitabine given by 24-h hepatic arterial infusion (HAI). PATIENTS AND METHODS: Patients with liver malignancies received gemcitabine by 24-h HAI, weekly x 3, every 4 weeks. On day 1 or day 8 of the first cycle, patients received one administration by 24-h intravenous infusion for pharmacokinetic comparison and to determine hepatic extraction. RESULTS: Thirteen patients received gemcitabine at the dose levels of 75, 135 and 180 mg/m(2). The MTD was 180 mg/m(2) with thrombocytopaenia as the dose-limiting toxicity. Pharmacokinetic analysis showed a significantly lower maximum gemcitabine plasma concentration (C(max): HAI, 26, 80 and 128 nM, respectively; IV, 229, 264 and 293 nM, respectively) and area under the plasma concentration-versus-time curve (AUC(0-24h): HAI, 386, 1247 and 2033 nmol x h/L, respectively; IV, 3526, 4818 and 5363 nmol x h/L, respectively) during HAI, compared with intravenous infusion (both P<0.001). Additionally, the mean hepatic extraction ratios of gemcitabine at the 75, 135 and 180 mg/m(2) dose level were 0.89, 0.75 and 0.55, respectively. Hepatic extraction decreased linearly with increasing dose. The C(max) and AUC(0-24h) of 2',2'-difluoro-2'-deoxyuridine, the deaminated product of gemcitabine, were similar for HAI and intravenous infusion. Seven patients had stable disease for a median duration of 9 months (range: 2-11 months). CONCLUSIONS: Gemcitabine given by 24-h HAI was well tolerated and resulted in significantly lower systemic gemcitabine plasma concentrations than intravenous infusion due to a relatively high hepatic extraction. PMID- 19556121 TI - Gain control in the visual thalamus during perception and cognition. AB - The thalamus has traditionally been thought to passively relay sensory information to the cortex. By showing that responses in visual thalamus are modulated by perceptual and cognitive tasks, recent fMRI and physiology studies have helped revise this view. The modulatory input to the visual thalamus derives from functionally distinct cortical and subcortical feedback pathways. These pathways enable the lateral geniculate nucleus and pulvinar to regulate the information transmitted to cortical areas according to cognitive requirements. Emerging evidence suggests that such regulation involves changing the degree of synchrony between neurons as well as changing the magnitude of thalamic activity. These findings support a role for the thalamus that extends as far as contributing to the control of visual attention and awareness. PMID- 19556123 TI - Characterization of start-up performance and archaeal community shifts during anaerobic self-degradation of waste-activated sludge. AB - Successful start-up strategy for anaerobic digestion of waste-activated sludge using internal inoculum and relationship between the shift of methanogenic community and the digester performance during start-up was investigated. Combination of TS control of inoculum and batch operation during early days enabled the successful start-up operation without serious volatile fatty acid accumulation, followed by the stable continuous operation. However, the propionate degradation was rate-limiting step during the batch operation. The results of real-time quantitative polymerase chain reaction analysis suggested that there was a correlation between the population of the genus Methanosarcina and the methane production rate coupled with acetate consumption during batch operation, and the results of terminal-restriction fragment length polymorphism (T-RFLP) revealed that the increasing intensity of T-RF peaks of hydrogenotrophic methanogens was associated with a decrease in the level of C3-acids. PMID- 19556124 TI - Application of life cycle assessment for an evaluation of wastewater treatment and reuse project--case study of Xi'an, China. AB - In order to illuminate the benefit of a wastewater treatment and reuse project, a life cycle assessment (LCA) model was proposed by combining the process-based LCA and the input-output based LCA in one framework and using energy consumption as the sole parameter for quantitative evaluation of the project. The life cycle consumption was evaluated mainly by life cycle inventory (LCI) analysis taking into account the construction phase, operation phase and demolishment phase of the project. For evaluating the life cycle benefit of treated water reuse, attention was paid to the decrease of secondary effluent discharge and water saving. As a result of comprehensive LCA analysis of a case project in Xi'an, China, it was understood that the life cycle benefit gained from treated wastewater reuse much surpassed the life cycle energy consumption. The advantage of wastewater treatment and reuse was well shown by LCA analysis using the proposed model. PMID- 19556126 TI - Structural and theoretical studies of [6-bromo-1-(4-fluorophenylmethyl)-4(1H) quinolinon-3-yl)]-4-hydroxy-2-oxo-3-butenoic acid as HIV-1 integrase inhibitor. AB - Ethyl [6-bromo-1-(4-fluorophenylmethyl)-4(1H)-quinolinon-3-yl]-4-hydroxy-2-oxo-3 butenoate 1 and [6-bromo-1-(4-fluorophenylmethyl)-4(1H)-quinolinon-3-yl)]-4 hydroxy-2-oxo-3-butenoic acid 2 were synthesized as potential HIV-1 integrase inhibitors and evaluated for their enzymatic and antiviral activity, acidic compound 2 being more potent than ester compound 1. X-ray diffraction analyses and theoretical calculations show that the diketoacid chain of compound 2 is preferentially coplanar with the quinolinone ring (dihedral angle of 0-30 degrees ). Docking studies suggest binding modes in agreement with structure-activity relationships. PMID- 19556125 TI - Discovery of a novel class of 2-mercaptohexanoic acid derivatives as highly active PPARalpha agonists. AB - A novel and robust scaffold for highly active PPARalpha agonists based on the 2 mercaptohexanoic acid substructure is presented. Systematic structural variation of the substitution pattern of the phenolic backbone yielded detailed SAR especially of ortho and meta substituents. We corroborated the importance of the sulfur atom as well as of the n-butyl chain for PPARalpha activity in the 2 mercaptohexanoic acid head group by preparation of carbon analogs and alpha unsubstituted derivatives. Compound 10 represents a low nano molar active PPARalpha activator with excellent selectivity towards PPARgamma. PMID- 19556127 TI - Synthesis and fungicidal activity of 3,5-dichloropyrazin-2(1H)-one derivatives. AB - We synthesized a family of 3,5-dichloropyrazin-2(1H)-one derivatives and assessed their in vitro fungicidal activity against Candida albicans. Compounds 11 and 20 were most active against C. albicans and induced accumulation of reactive oxygen species in this pathogen. Using a genome-wide approach in the yeast Saccharomyces cerevisiae, we demonstrated that genes involved in vacuolar functionality and DNA related functions play an important role in cellular mechanisms underlying the fungicidal activity of these compounds. PMID- 19556128 TI - Tetrahydrocarbazole amides with potent activity against human papillomaviruses. AB - Synthesis of a series of tetrahydrocarbazole amides with potent activity against human papillomaviruses is described. Synthetic approaches allowing for variation of the substitution pattern of the tetrahydrocarbazole and the amide are outlined and resulting changes in antiviral activity and certain developability parameters are highlighted. Several compounds with in vitro antiviral activity (W12 antiviral assay) in the single digit nanomolar range were identified and N-[(1R) 6-chloro-2,3,4,9-tetrahydro-1H-carbazol-1-yl]-2-pyridinecarboxamide was selected for further evaluation. PMID- 19556129 TI - Immunohistochemical analysis of calpain 3: advantages and limitations in diagnosing LGMD2A. AB - Immunoblot is currently the preferred laboratory test to assist the diagnosis of limb-girdle muscular dystrophy (LGMD) 2A (calpainopathy). To assess whether immunohistochemistry may offer a reliable alternative screening we used two antibodies, Calp3-2C4 (exon 1) and Calp3-12A2 (exon 8), to label blots and sections of skeletal muscle from controls and patients with LGMD2A and other muscle diseases. In LGMD2A muscle biopsies a high degree of concordance was found with Calp3-2C4: labelling on sections was absent in patients with no bands on immunoblot and detected in those where CAPN3 bands were seen. Calp3-12A2 results were less consistent, with most samples retaining labelling. Interestingly, CAPN3 was found in all muscle sections from disease control patients irrespective of its detection on immunoblot. Our results show that immunohistochemistry with Calp3-2C4 has a similar pickup rate of LGMD2A as immunoblot and it may therefore be useful for distinguishing the majority of genuine CAPN3 defects from secondary protein reduction. However immunoblot is still needed when CAPN3 is present on sections to show secondary CAPN3 reduction and to identify LGMD2A with variable reduction of CAPN3 bands. PMID- 19556130 TI - White matter lesions in spastic paraplegia. PMID- 19556131 TI - Rhabdomyolysis caused by tocolytic therapy with ritodrine hydrochloride. AB - We report a case of rhabdomyolysis with severe generalized weakness and muscle pain after administration of ritodrine hydrochloride, in a pregnant patient without history of neuromuscular disease. Laboratory tests showed an increase of blood CK value and myoglobinuria. An electromyography was performed, revealing a typical myogenic pattern and diffuse denervation activity. Muscular biopsy allowed to rule out inflammatory and metabolic myopathy. After delivery, the patient underwent intensive rehabilitation with progressive improvement of her clinical situation, until complete recovery. Three months later, both neurological assessment and CK levels were normal. This case highlights that rhabdomyolysis has to be considered even in patient treated with ritodrine alone and without history of neuromuscular disease. Therefore, muscular symptoms and CK levels should be monitored in women treated with ritodrine for premature labour. PMID- 19556132 TI - Percutaneous transpedicular vertebroplasty with polymethyl methacrylate for pathological fracture of the spine. AB - We aimed to evaluate the safety and therapeutic efficacy of percutaneous transpedicular vertebroplasty (PVP) using polymethyl methacrylate (PMMA) in patients with symptomatic metastatic spine lesions. We included 31 patients in this retrospective study who were treated with PMMA from 2003 to 2005 for intractable pain due to metastatic spine lesions. The types of cancer (and numbers of patients) included: lung cancer (9), breast cancer (7), gastrointestinal (GI) tract cancers (5), hepatobiliary malignancies (3), and other types of cancer (7). All patients received vertebroplasty, resulting in 41 treatments (16 in thoracic, 25 in lumbar spine). Preoperative and postoperative visual analog scale (VAS) scores for pain were measured in all patients. Image studies including contrast-enhanced MRI were performed in all patients. Results showed characteristic metastatic lesions. Suspicious lesions were further confirmed as malignant by a bone scan, a positron emission tomography (PET) scan, and pathological exam. Vertebroplasty resulted in complete or partial pain relief in 29 patients (95%), and provided no pain relief in 2 patients (5%). The mean preoperative VAS score of 8.9 (+/-0.93) was higher than the mean postoperative VAS score (2.6+/-1.71). Metastatic spine lesions were most common in lung and breast cancer patients and these lesions were located more often on segments T12 to L2 (53.6%). Patients with malignancy of hepatobiliary origin did not show improvement in pain scores as dramatically as patients with other types of malignancies, although only a few cases were included in this study. No patients experienced worsening of symptoms or suffered from vertebroplasty complications. We conclude that vertebroplasty is a safe, effective, and simple treatment for the management of intractable spinal pain due to metastases. PMID- 19556133 TI - Post-traumatic intradiploic leptomeningeal cyst of the posterior fossa in an adult. AB - Intradiploic cysts in the posterior fossa are rare. We report a post-traumatic intradiploic leptomeningeal cyst in an adult and review the diagnosis and pathogenesis of this lesion. A 28-year-old woman presented with a headache and a hard mass in the occipital region. She had a history of head injury as she had fallen from a height at the age of 18 months. CT scans and MRI revealed an expanding intradiploic cyst with the density of cerebrospinal fluid (CSF) and thinning of the inner and outer tables. A bony defect of the inner table connecting to the cisterna magna was also visualized. At surgery, we detected free communication of the CSF between the cystic cavity and the subarachnoid space of the posterior fossa via a defect in the dura and inner table. The dural defect was sutured primarily and covered with the autogenous outer table. An intradiploic cyst of the occipital bone can be detected in adults and might be caused by remote head injuries during childhood. Prompt surgical repair of the dural and bony defect in an adult patient has an excellent prognosis. PMID- 19556135 TI - Venous air emboli from intravenous catheterization: a report of iatrogenic intravascular pneumocephalus. AB - Pneumocephalus in the absence of a recent intracranial or intrathecal procedure is a significant radiographic finding and may be clinically relevant. Pneumocephalus secondary to intravenous catheterization may be a more common finding than previously expected. Although pneumocephalus is often associated with pathological conditions, recognition of a characteristic pattern of intravenous pneumocephalus in the presence of intravenous catheterization may aid the clinician in determining a patient's underlying condition. This unexpected radiographic finding should not necessarily be cause for alarm, and there is no evidence that intravenous pneumocephalus alone is harmful. We present a patient with intravenous pneumocephalus and a review of the literature. PMID- 19556134 TI - Tumour angiogenesis: its mechanism and therapeutic implications in malignant gliomas. AB - Angiogenesis is a key event in the progression of malignant gliomas. The presence of microvascular proliferation leads to the histological diagnosis of glioblastoma multiforme. Tumour angiogenesis involves multiple cellular processes including endothelial cell proliferation, migration, reorganisation of extracellular matrix and tube formation. These processes are regulated by numerous pro-angiogenic and anti-angiogenic growth factors. Angiogenesis inhibitors have been developed to interrupt the angiogenic process at the growth factor, receptor tyrosine kinase and intracellular kinase levels. Other anti angiogenic therapies alter the immune response and endogeneous angiogenesis inhibitor levels. Most anti-angiogenic therapies for malignant gliomas are in Phase I/II trials and only modest efficacies are reported for monotherapies. The greatest potential for angiogenesis inhibitors may lie in their ability to combine safely with chemotherapy and radiotherapy. PMID- 19556136 TI - Frontotemporal dementia and motor neurone disease: overlapping clinic pathological disorders. AB - Advances in genetics and pathology have supported the idea of a continuum between frontotemporal dementia (FTD) and motor neurone disease (MND), which is strengthened by the discovery of the trans-activating responsive (Tar) sequence DNA binding protein (TDP-43) as a key component in the underlying pathology of FTD, FTD-MND and sporadic and familial MND patients. MND is a multisystem disorder associated with cognitive and behavioural changes which in some instances reaches the criteria for FTD, while a proportion of patients with FTD develop frank MND. We review the overlap between FTD and MND, emphasizing areas of controversy and uncertainty. PMID- 19556137 TI - Gait and muscle activation changes in men with knee osteoarthritis. AB - The aim was to examine the biomechanics of level- and stair-walking in men with knee osteoarthritis (OA) at different pre-determined gait speeds and to compare the results with those obtained from healthy control subjects. Special emphasis was placed on the estimation of joint loading. Fifty-four men with knee OA (50-69 years) and 53 healthy age- and sex-matched controls were enrolled in the study. The participants walked barefoot in the laboratory (1.2 m/s+/-5%), corridor (1.2; 1.5 and 1.7 m/s+/-5%), and climbing and coming down stairs (0.5 and 0.8 m/s+/-5%) separately. Joint loading was assessed with skin mounted accelerometers (SMAs) attached just above and below the more affected knee joint. The 3-D ground reaction forces (GRFs) and muscle activation with surface-electromyography (EMG) from vastus medialis (VM) and biceps femoris (BF) were also measured simultaneously. There were no differences in SMA variables between groups during level-walking, but maximal loading rate (LR(max)) was higher bilaterally in the controls (P<.05). Patients loaded their lower extremity more forcefully especially during stair descent at faster speed. The distinctions in muscle activation both at level- and stair ambulation in VM and BF muscles revealed that the patients used different strategies to execute the same walking tasks. It is concluded that the differences in measured SMA and GRF parameters between the knee OA patients and the controls were only minor at constant gait speeds. It is speculated that the faster speeds in the stair descent subjected the compensatory mechanisms to the maximum highlighting the differences between groups. PMID- 19556138 TI - Anti-tumor and anti-angiogenic activity of novel hydantoin derivatives: Inhibition of VEGF secretion in liver metastatic osteosarcoma cells. AB - A series of new azaspiro bicyclic hydantoin derivatives has been designed and synthesized. Initially, the anti-proliferative effect of the hydantoin derivatives was evaluated against human ovarian cancer cells (SKOV-3 and OVSAHO) and murine osteosarcoma cells (LM8 and LM8G7). Among the tested compounds, 8-(3 fluorobenzyl)-1'-(4-(methylsulfonyl)benzyl)-8-azaspiro[bicyclo[3.2.1]octane-3,4' imidazolidine]-2',5'-dione (7h) and 8-(3,4-difluorobenzyl)-1'-(4 (methylsulfonyl)benzyl)-8-azaspiro[bicyclo[3.2.1]octane-3,4'-imidazolidine]-2',5' dione (7i) showed a significant anti-proliferative activity against the OVSAHO and LM8G7 cells. The real-time monitoring of the effect of the compounds 7h and 7i against the proliferation of LM8G7 was revealed that resulting IC(50) values were 102microM and 13microM, respectively. We reasoned that the presence of fluorine atom at the 3rd position of the phenyl ring of the hydantoin side chain may determine the potency of the molecule. Furthermore, the compound 7i inhibited the tube formation of the mouse endothelial cells. Finally, the treatment of the compound 7i against the proliferation of LM8G7 cells demonstrated the down regulation of the secretion of VEGF, indicate the potential angioinhibitory effects. In conclusion, our findings demonstrate the suppression of the secretion of VEGF by LM8G7 cells by the compound 7i might contribute at least in part to the antitumor action. PMID- 19556139 TI - Structure and ultrastructure of the spermatozoa of Trypoxylon (Trypargilum) albitarse Fabricius 1804 (Hymenoptera: Apoidea: Crabronidae). AB - The ultrastructure of the spermatozoa of Trypoxylon (Trypargilum) albitarse is described here for the first time within this genus. Testes and seminal vesicles were dissected and processed for transmission electron microscopy. In the testicular follicles, the spermatids are arranged in a maximum number of 32 for each cyst. The spermatozoa are slender and they measure approximately 150 microm in length. The head is about 17 microm long and is formed by the acrosome and the nucleus. The flagellum consists in an axoneme, two mitochondrial derivatives, two accessory bodies and, at the nucleus-flagellum transition, a symmetric centriolar adjunct. The axoneme presents the typical 9+9+2 microtubule pattern. In the terminal region, the central microtubules and nine doublets finish first, followed by the accessory microtubules. Both mitochondrial derivatives begin together and are inserted in the base of the centriolar adjunct. Along the middle region, the larger derivative has almost twice the area of the smaller one and includes a discrete paracrystalline region. At the tip, the smaller derivative ends before the larger one and both before the axoneme. The characters derived from the ultrastructure of the spermatozoa of T. albitarse show synapomorphies shared with the Apoidea and present characters that are probably apomorphic for the Crabroninae subfamily. PMID- 19556140 TI - Generation of asparagine-linked glycan structure databases and their use. AB - Asparagine linked glycans (N-glycans) are important in biological processes. Yet, their structural complexity and lack of databases hinder progress in glycomics and glycobiology. We present a way for in silico generation of very large N glycan structure databases and their use in high throughput composition and primary structure determination of N-glycans attached to peptides, based on CID (collision induced dissociation) MS/MS (tandem mass spectrometric data). The database and the integrated search engine is called Glyquest and is available to the glycomics community. PMID- 19556141 TI - Decreased nitric oxide products in the urine of patients undergoing cardiac surgery. PMID- 19556142 TI - Pro: Tight perioperative glycemic control. PMID- 19556143 TI - Real-time three-dimensional transesophageal echocardiography and a congenital bilobar left atrial appendage. PMID- 19556144 TI - Revised Cardiac Risk Index (Lee) and perioperative cardiac events as predictors of long-term mortality in patients undergoing endovascular abdominal aortic aneurysm repair. AB - OBJECTIVE: To determine if the Revised Cardiac Risk Index (Lee) is useful for stratification of patients by risk of both perioperative cardiac morbidity and long-term all-cause mortality in the setting of endovascular repair of abdominal aortic aneurysms. DESIGN: This study was designed as a retrospective review. SETTING: It was conducted at a single academic medical institution. PARTICIPANTS: The analysis included 225 patients with abdominal aortic aneurysms admitted to the authors' institution from 1999 to 2006. INTERVENTIONS: All patients underwent endovascular aortic aneurysm repair. MEASUREMENTS AND MAIN RESULTS: Data were collected from medical records, office charts, and physician quality-assurance databases. There were no in-hospital cardiac deaths. The major adverse cardiac event rate in the perioperative period was 6.2%. Long-term all-cause mortality was 23%. Univariate analysis showed that a history of coronary artery disease (CAD) (likelihood ratio [LR] = 8.7, p = 0.023), history of congestive heart failure (LR = 4, p = 0.042), and a Revised Cardiac Risk Index (RCRI) > or =3 (LR = 8.6, p = 0.004) were significant predictors for perioperative major adverse cardiac events. A history of CAD (LR = 10.7, p = 0.002), echocardiographic evidence of myocardial infarction (LR = 8.5, p = 0.006), exercise tolerance of only 1 block (LR = 8.4, p = 0.005), RCRI > or =3 (LR = 5.6, p = 0.022), and perioperative cardiac events (LR = 15.9, p < 0.0001) were significantly associated with long-term all-cause mortality. Perioperative cardiac events remained highly significant in predicting long-term mortality within the RCRI > or =3 subgroup (LR = 6.1, p = 0.019). CONCLUSIONS: The results of this study confirm that long-term mortality remains high after endovascular repair of abdominal aortic aneurysms. The Lee index may be a useful tool for stratification of high-risk patients from both a short- and long-term perspective in the setting of endoluminal graft repair. PMID- 19556145 TI - Fewer rotator cuff tears fifteen years after arthroscopic subacromial decompression. AB - BACKGROUND: A successful clinical result is reported in 75% to 85% of impingement patients after arthroscopic subacromial decompression. The result is maintained over time, but few studies have investigated the integrity of the rotator cuff in these patients. MATERIALS AND METHODS: Using ultrasonography, we examined the integrity of the rotator cuff in 70 patients 15 years after arthroscopic subacromial decompression. All patients had an intact rotator cuff at the index procedure. RESULTS: Tendons were still intact in 57 patients (82%), 10 (14%) had partial-thickness tears, and 3 (4%) had full-thickness tears. DISCUSSION: The total number of 18% tears (partial and full thickness) in this study, including patients clinically diagnosed with subacromial impingement at a mean age of 60 years, is unexpectedly low compared with 40% degenerative tears reported in asymptomatic adults of the same age. CONCLUSION: Arthroscopic subacromial decompression seems to reduce the prevalence of rotator cuff tears in impingement patients. This appears attributable to elimination of extrinsic factors such as mechanical wear and bursitis. The potential effect of surgery on intrinsic cuff degeneration is unknown, but intrinsic factors may explain tears still developing despite decompression. LEVEL OF EVIDENCE: Level III, therapeutic study. PMID- 19556146 TI - Surgical treatment for osteochondritis dissecans of the humeral capitellum. AB - BACKGROUND: Surgical treatments have been selected to treat advanced osteochondritis dissecans (OCD) of the humeral capitellum. PURPOSE: To evaluate the clinical results of surgical treatments for advanced capitellar OCD lesion and to clarify problems of poor clinical results. METHODS: Cases were reviewed for 27 patients with advanced OCD lesions treated operatively. All patients were male baseball players, with a mean of 13.3 years at the time of surgery. Drilling only was performed for 3 patients, fragment fixation for 13 patients, and removal of the detached fragment with drilling for four patients. Since 2004, reconstruction of the articular surface with use of osteochondral autograft from the rib has been performed for 7 patients with osteochondral defect. Mean follow up was 37.4 months. Follow-up assessment included modified elbow rating system, evaluation of radiographs, and return to sports. RESULTS: Mean subjective score improved significantly from 70 to 96 postoperatively. Mean objective score improved significantly 71 to 81 postoperatively. A postoperative return to baseball was achieved by 25 patients. On the final radiographs, 4 patients showed flattening of >70% of the capitellum or degenerative changes, including insufficient remodeling of the lateral margin of the capitellum. Although patients could return to baseball, postoperative total arc of the elbow was decreased compared with the preoperative total arc. CONCLUSION: Surgical treatments were useful to restore advanced OCD lesions. Our results suggest that reconstruction of the lateral margin of the capitellum is important for achieving good clinical results. LEVEL OF EVIDENCE: Level 4. PMID- 19556147 TI - Measurement of the acromiohumeral interval on standardized anteroposterior radiographs: a prospective study of observer variability. AB - BACKGROUND: An acromiohumeral interval narrower than 6 mm has been considered pathologic and strongly indicative for rotator cuff tears by numerous authors. This prospective study assessed interobserver and intraobserver variability in the radiographic measurement of the acromiohumeral interval. MATERIAL AND METHODS: Five board-certified orthopedic surgeons independently reviewed 58 blinded, standardized anteroposterior shoulder radiographs. The acromiohumeral interval was measured in millimeters. The 5 investigators classified each image a second time in random order. RESULTS: After the same 58 radiographs had been evaluated by the 5 investigators at both examination time points, no significant differences were noted in the interobserver and intraobserver measurements (P < .05). The respective maximum interobserver and intraobserver differences were 4 and 3 mm (range, 0-4 mm). CONCLUSION: The assessment of the acromiohumeral interval using standardized anteroposterior radiographs is a reliable and reproducible method of measurement. LEVEL OF EVIDENCE: Level 1; Investigating a diagnostic test. PMID- 19556148 TI - Survivorship of the humeral component in shoulder arthroplasty. AB - BACKGROUND: Loosening of the humeral component is rarely a cause for revision. This study was conducted to determine long-term survivorship of humeral components and investigate the risk factors associated with humeral component removal or revision. MATERIALS AND METHODS: From 1984 to 2004, 1423 patients underwent 1584 primary Neer and Cofield shoulder arthroplasties. The Kaplan-Meier method was used to estimate implant survival. Cox proportional hazards regression was used to assess the effects of age, gender, etiology of the disease, surgery type (hemi vs total), fixation (cemented vs noncemented), and the humeral component design (Neer II, Cofield 1 or 2) with survival free of revision or removal of the humeral component. RESULTS: There were 108 revisions and 17 removals of the humeral component. Estimates of survivorship free of revision or removal of the humeral component for any reason was 94.8% (95% confidence interval [CI], 93.6-96.0) at 5 years, 92.0% (95% CI, 90.4-93.6) at 10 years, 86.7% (95% CI, 84.2-89.4) at 15 years, and 82.8% (95% CI, 78.5-87.5) at 20 years. Younger age, male gender, replacement due to posttraumatic arthritis, an uncemented component, and use of a metal-backed glenoid component increased the likelihood of humeral component failure. CONCLUSIONS: The need for revision of the humeral component is commonly related to glenoid or glenoid component issues. Patient and diagnostic factors play a role in implant survival; implant type and method of fixation are less important. LEVEL OF EVIDENCE: Level 4; Case series, treatment study. PMID- 19556149 TI - Loss of the sclerotic line of the glenoid on anteroposterior radiographs of the shoulder: a diagnostic sign for an osseous defect of the anterior glenoid rim. AB - BACKGROUND: The integrity of the glenoid defines the surgical treatment in anterior shoulder instabilities. The reliability of plain radiographs to detect anterior glenoid rim deficiencies was determined. MATERIALS AND METHODS: The anterior sclerotic glenoid line (SGL) was assessed on anteroposterior radiographs of 86 shoulders (34 anterior instabilities, 15 posterior instabilities, 37 stable) and compared with computed tomography (CT) scans (gold standard). A loss of the SGL (LSGL) was defined as a positive LSGL sign. RESULTS: On CT scans, 25 of 34 shoulders (74%) with anterior instabilities showed a defect of the anterior glenoid rim. No defects were found in shoulders without anterior instabilities. LSGL correctly predicted an anterior glenoid rim lesion in 16 (examiner A) or 14 (examiner B) of the 25 anterior instabilities (sensitivity, 64% and 56%), without a false-positive diagnosis (specificity, 100%). CONCLUSION: The LSGL on anteroposterior radiographs is a moderately sensitive but highly specific finding for anterior glenoid rim defects. LEVEL OF EVIDENCE: Level 4; Diagnostic study, case control study. PMID- 19556150 TI - Effectiveness of multidetector computed tomography arthrography for the diagnosis of shoulder pathology: comparison with magnetic resonance imaging with arthroscopic correlation. AB - HYPOTHESIS: This study evaluated the diagnostic efficacy of computed tomography arthrography (CTA) in the assessment of various shoulder pathologies with arthroscopic correlation. We hypothesized that CTA would be cost-effective and effectively comparable with magnetic resonance arthrography (MRA) for assessing labral detachments and full-thickness rotator cuff tears. MATERIALS AND METHODS: A musculoskeletal radiologist interpreted CTAs for 78 patients and MRAs for 70 patients. Each imaging study was evaluated for the presence of bony (Hill-Sachs) or labral (Bankart or superior labrum anteroposterior [SLAP]) lesions, and rotator cuff disorder (full- or partial-thickness tears). All patients subsequently underwent arthroscopic surgery. Detailed arthroscopic findings were reported and compared with CTA and MRA findings. The sensitivity, specificity, kappa coefficients, and the area under the receiver operating characteristic (AUROC) curve were calculated. RESULTS: The sensitivity, specificity, and agreement were comparable in each imaging study for Bankart, SLAP, and Hill-Sachs lesions, and full-thickness rotator cuff tears, but those of CTA were significantly lower than MRA for partial-thickness cuff tears. The AUROC curve for CTA and MRA were not significantly different for any of the pathologies, except partial-thickness cuff tears. CONCLUSIONS: Our data suggest that CTA is a cost-effective, useful method in the preoperative evaluation of labral abnormalities, such as Bankart and SLAP lesions. It may also be useful for the detection of full-thickness rotator cuff tears. LEVEL OF EVIDENCE: Level I; Diagnostic study. PMID- 19556151 TI - Continuous bilateral infusion of GABA in the dorsomedian nucleus of the thalamus elevates the generalized seizure threshold in amygdala-kindled rats. AB - In order to assess the role of continuous intracerebral infusion of GABA over the propagation of generalized seizures from the amygdala, Wistar rats were subjected to a kindling procedure at the left basolateral amygdala. Subsequently, they were implanted with miniosmotic pumps filled with 100mg/mL of GABA in saline, connected to catheters whose tips were placed bilaterally at both dorsomedian nuclei of the thalamus (DMNT). The threshold intensity to provoke local afterdischarges (ADT) and generalized seizures (GST) were measured before, during and after GABA infusion, as well as seizure intensity and signs of ataxia and sedation. While there was no observed variation on ADT, the median GST was significantly increased during, but not after infusion of GABA (P=0.047, compared to the preinfusion value). Seizure intensity was not changed. No signs of neurologic side effects were recorded. These data emphasize the role of DMNT in the generalization of seizures originated at the amygdala. PMID- 19556152 TI - Long-term outcome of salvage high-dose chemotherapy in patients with germ cell tumor with poor prognostic features. AB - OBJECTIVE: High-dose chemotherapy (HDCT) represents an option as salvage treatment for patients with resistant/refractory germ cell tumor (GCT). The objective of this retrospective analysis was to evaluate the long-term results of a single-center experience with salvage HDCT for GCT patients, and to validate the prognostic model proposed by Einhorn and colleagues [9]. MATERIALS AND METHODS: Between 1986 and 2003, 100 GCT patients received salvage HDCT consisting of high-doses of carboplatin, etoposide +/- cyclophosphamide, or ifosfamide. Twenty-four patients underwent a second HDCT cycle, and in 1 case, a third cycle was given with a median interval time of 6 weeks (range, 5-10). RESULTS: With a median follow-up of 8 years (range, 3-17); 6 of 32 (19%) patients with resistant GCT and 1 of 19 (5%) patients with cisplatin-refractory disease have been continuously disease-free, while none of the 16 patients with absolutely cisplatin-refractory GCT were alive at 1 year from HDCT treatment. In the PBPC era, HDCT appeared to be inapplicable in 32% of patients, mainly due to progressive disease during the induction/mobilizing phase. The prognostic model by Einhorn et al. for tandem HDCT did categorize our patients treated with a single HDCT cycle or low-dose intensity regimens in a very similar manner, but with inferior overall results. CONCLUSIONS: Long-term results with a single HDCT cycle or a low dose-intensity multicycle HDCT regimen remained poor in patients with adverse prognostic features. The tandem HDCT regimen represents a major option for refractory GCTs and relapsed tumors in third-line or later therapy, while a single course of HDCT should be abandoned for these patients. PMID- 19556153 TI - Comparison of 2002 TNM nodal status with lymph node density in node-positive patients after radical cystectomy for bladder cancer: analysis by the number of lymph nodes removed. AB - OBJECTIVE: Because 2002 TNM pathologic nodal (pN) status was established in patients from whom a relatively small number of lymph nodes had been removed, it is necessary to validate this staging system in current clinical practice, in which the removal of more lymph nodes is recommended during radical cystectomy and pelvic lymphadenectomy. We assessed the ability of lymph node density (LND) and 2002 TNM pathologic nodal (pN) status to predict disease-specific survival (DSS) in node-positive patients after radical cystectomy for bladder cancer, and investigated whether these factors were affected by the number of lymph nodes removed during pelvic lymphadenectomy. MATERIALS AND METHODS: We retrospectively evaluated outcomes in 130 patients with nodal metastases after radical cystectomy performed between 1989 and 2006. Patients were divided into 2 subgroups based on the median number of lymph nodes removed, those with <15 and those with >= 15 lymph nodes removed. The effect of several variables on DSS was assessed. RESULTS: The overall 5-year DSS rate was 38.5%. Multivariate analysis showed that in the entire cohort, LND (HR = 2.28, 1.04-5.03, P = 0.041) and the use of adjuvant chemotherapy (HR = 2.68, 1.42-5.06, P = 0.002) were significant predictors of DSS. In patients with <15 lymph nodes removed, pN status (HR = 5.19, 1.24-21.75, P = 0.024) and use of adjuvant chemotherapy (HR = 6.23, 2.32 16.73, P < 0.001) were independent predictors of DSS. In patients with >= 15 lymph nodes removed, however, only LND (HR = 4.08, 1.10-15.10, P = 0.036) was a predictor of DSS. CONCLUSIONS: LND was an independent predictor of DSS in node positive patients. However, when small numbers of lymph nodes were removed, TNM pN status was a better predictor than LND. These findings suggest that abilities of TNM pN status and LND in node-positive patients to predict DSS could be affected by the total number of lymph nodes removed. PMID- 19556155 TI - Localized donor cells in brain of a Hunter disease patient after cord blood stem cell transplantation. AB - The efficacy of hematopoietic stem cell transplantation (HSCT) for Hunter disease (deficiency of iduronate-2-sulfatase, IDS) remains unclear. We treated a 6-year old male suffering from a severe type of Hunter disease with cord blood stem cell transplantation (CBSCT); however, he died at 10 months post-therapy due to a laryngeal post-transplantation lymphoproliferative disorder. During the follow-up period after CBSCT, his hyperactivity, estimated mental age, and brain MR findings had not improved. We assessed the efficacy of CBSCT by biochemical and pathological analyses of the autopsied tissues. There were many distended cells with accumulated substrate in the brain, but not in the liver. IDS enzyme activity in the cerebrum remained very low, although that in the liver reached about 40% of the normal control level. However, a variable number of tandem repeats analyses demonstrated a weak donor-derived band not only in the liver but also in the cerebrum. Furthermore, IDS-immunoreactivity in the liver was recognized broadly not only in Kupffer cells but also in hepatocytes. On the other hand, IDS-immunoreactivity was recognized exclusively in CD68-positive microglia/monocytes in the patient's brain; whereas that in the normal brain was also detected in neurons and oligodendrocytes. These donor-derived IDS-positive cells were predominantly localized in perivascular spaces and some of them were evidently present in the brain parenchyma. The efficacy of CBSCT was judged to be insufficient for the brain at 10 months post-therapy. However, the pathological detection of donor-derived cells in the brain parenchyma suggests the potential of HSCT for treatment of neurological symptoms in Hunter disease. This is the first neuropathological report documenting the distribution of donor-derived cells in the brain after CBSCT into a Hunter disease patient. PMID- 19556154 TI - Bladder cancer after nephroureterectomy in patients with urothelial carcinoma of the upper urinary tract. AB - OBJECTIVES: To determine the independent risk factors of bladder recurrence in patients with upper urinary tract (UUT) urothelial carcinoma (UC). MATERIALS AND METHODS: A total of 181 patients with UUT-UC were enrolled in this study. Their median age was 63 years (range 36-90), and median follow-up after total nephroureterectomy was 37.5 months (range 1.0-174.0). The end-point of this study was defined as the initial intravesical recurrence of UC. RESULTS: Of the 181 patients, 64 (35.4%) developed subsequent bladder tumors at a median interval of 6.3 months (range 1.7-50.1) after initial treatment. By univariate analysis, a previous bladder tumor history (P = 0.046) and tumor necrosis (P < 0.001) were found to have a significant prognostic impact on bladder tumor-free survival in patients with superficial UUT-UC, whereas surgical margin status (P = 0.045) and the use of adjuvant chemotherapy (P = 0.003) were found to be prognostic factors for bladder tumor-free survival in those with invasive UUT-UC. However, by multivariate analysis, only tumor necrosis (P = 0.012, relative risk = 6.512) was found to have a significant impact on intravesical recurrence in patients with superficial UUT-UC. However, surgical margin status (P = 0.007, relative risk = 5.846) and the use of adjuvant chemotherapy (P = 0.001, relative risk = 0.223) were retained as independent predictors of bladder tumor survival in those with invasive UUT-UC. CONCLUSIONS: Our findings may be useful in patients with UUT-UC who may require more stringent follow-up by cystoscopy to detect bladder tumors. PMID- 19556156 TI - Infiltrative lymphocytic mural folliculitis: a histopathological reaction pattern in skin-biopsy specimens from cats with allergic skin disease. AB - This study was conducted to determine the prevalence of infiltrative lymphocytic mural folliculitis (ILMF) in skin-biopsy specimens from 354 cats with various inflammatory dermatoses and from 33 cats with normal skin. Although ILMF was present in 33/47 dermatoses evaluated, the prevalence of ILMF in allergic dermatoses (116/172 cats; 67%) was significantly greater than that in non allergic dermatoses (61/182 cats; 33%). Cats with allergic dermatoses had a 4.1 times greater odds of having ILMF than cats with non-allergic dermatoses. ILMF was not observed in any of the normal skin specimens. PMID- 19556157 TI - Paracoccus yeei: a new unusual opportunistic bacterium in ambulatory peritoneal dialysis. AB - Paracoccus yeei was identified as the etiologic agent of peritonitis in an ambulatory peritoneal dialysis patient. While the old biochemical identification kits are not able to identify this species, the new colorimetric VITEK 2 GN card correctly identified this isolate in 7hours. Its identity was confirmed by sequencing of the 16S rRNA gene. PMID- 19556158 TI - [Acute carpal tunnel syndrome after trauma: a series of 26 cases]. AB - The aim of the study is to evaluate the results of the median nerve release at wrist as a part of acute carpal tunnel syndrome. This is a retrospective study about 26 cases of acute carpal tunnel syndrome following isolated injury of the wrist. Clinical evaluation of the median nerve function was performed before and after release. A classical median nerve release was performed in emergency eventually associated with stabilization of bone lesions. The population included was young, the mean age was 26 (range 4-52) years, frequently male (sex-ratio 3.5:1). The wrist trauma was direct in 75% of all cases and oftentimes with high velocity. The median nerve was deficient in the first examination. The average time from injury to surgery range from three hours to five days. The release was performed in emergency. The latest follow-up showed a complete recovery of the nerve's function in 85% and an incomplete in the rest (15%).The mean prognosis factor was the release time. Wrist trauma particularly with high velocity should lead to evaluation of median nerve function and to an emergency release when acute median neuropathy appeared. PMID- 19556159 TI - High energy biofuel from endophytic fungi? AB - A paper entitled 'The production of myco-diesel hydrocarbons and their derivatives by the endophytic fungus Gliocladium roseum (NRRL 50072)' (Strobel et al., Microbiology 154, 3319-3328, 2008) demonstrated for the first time that a fungal endophyte can produce volatile hydrocarbons, which are major constituents of diesel fuel. The possible implications of this finding with regards to potential commercial applications and the ecological role of volatiles in endophyte-host relationships are discussed. PMID- 19556160 TI - Next generation therapeutics. PMID- 19556161 TI - Experimental and theoretical comparative studies on two 2-pyrazoline derivatives. AB - Two 2-pyrazoline derivatives of 1-phenyl-3-(4-methylphenyl)-5-phenyl-2-pyrazoline (1) and 1-phenyl-3-(4-methylphenyl)-5-(2,4-dichlorophenyl)-2-pyrazoline (2) have been synthesized and characterized by elemental analysis, IR, UV-vis and fluorescence spectroscopy. The crystal structure of 2 has been determined by X ray single crystal diffraction. For the two compounds, density functional theory (DFT) calculations of the structures and natural population atomic charge analysis have been performed at B3LYP/6-311G** level of theory. By using TD-DFT method, electron spectra of 1 and 2 have been predicted, which suggests the B3LYP/6-311G** method can approximately simulate the electron spectra for the system presented here. Comparative studies on 1 and 2 indicate that the change of substituent in 5-phenyl ring of pyrazoline ring influences the peak location and intensity in electronic and fluorescence spectra. PMID- 19556162 TI - Format change of a laboratory test order form affects physician behavior. AB - PURPOSE: Our study was designed to find whether a change in physician ordering of laboratory testing could be obtained by the simple strategy of changing the set up of the check-box laboratory order form that is embedded in a computerized medical record. METHODS: This prospective intervention study was undertaken in Maccabi Healthcare Services, a Preferred Provider Organization that has used a computerized medical record since 1992. We examined data from 865 primary healthcare physicians over 3 years. In May 2005 we changed the order form and reduced the number of tests that can be ordered using a check-box form from 51 to 26. Twenty-seven tests were removed from the form and two tests were added. The total number of laboratory test orders and the median rate of test orders per visit to physician during each of the study periods were calculated separately for each test. RESULTS: Tests that were added to the computerized laboratory order form showed an increase of 60.7% in the first year and a further 90% increase in the following year. For the unchanged tests the percentage changes over the same periods were +18.4% and -22.4%. For the deleted tests the change was -27% and -19.2% for the respective years. CONCLUSIONS: Changes in format of laboratory test order forms can change physician test ordering and may be useful together with other interventions to improve appropriateness of laboratory testing. A thoughtfully built test ordering form can reinforce clinical guidelines for the performance of some preventive testing and follow-up. PMID- 19556163 TI - Effects of bariatric surgery on cancer incidence in obese patients in Sweden (Swedish Obese Subjects Study): a prospective, controlled intervention trial. AB - BACKGROUND: Obesity is a risk factor for cancer. Intentional weight loss in the obese might protect against malignancy, but evidence is limited. To our knowledge, the Swedish Obese Subjects (SOS) study is the first intervention trial in the obese population to provide prospective, controlled cancer-incidence data. METHODS: The SOS study started in 1987 and involved 2010 obese patients (body mass index [BMI] >or=34 kg/m(2) in men, and >or=38 kg/m(2) in women) who underwent bariatric surgery and 2037 contemporaneously matched obese controls, who received conventional treatment. While the main endpoint of SOS was overall mortality, the main outcome of this exploratory report was cancer incidence until Dec 31, 2005. Cancer follow-up rate was 99.9% and the median follow-up time was 10.9 years (range 0-18.1 years). FINDINGS: Bariatric surgery resulted in a sustained mean weight reduction of 19.9 kg (SD 15.6 kg) over 10 years, whereas the mean weight change in controls was a gain of 1.3 kg (SD 13.7 kg). The number of first-time cancers after inclusion was lower in the surgery group (n=117) than in the control group (n=169; HR 0.67, 95% CI 0.53-0.85, p=0.0009). The sex treatment interaction p value was 0.054. In women, the number of first-time cancers after inclusion was lower in the surgery group (n=79) than in the control group (n=130; HR 0.58, 0.44-0.77; p=0.0001), whereas there was no effect of surgery in men (38 in the surgery group vs 39 in the control group; HR 0.97, 0.62 1.52; p=0.90). Similar results were obtained after exclusion of all cancer cases during the first 3 years of the intervention. INTERPRETATION: Bariatric surgery was associated with reduced cancer incidence in obese women but not in obese men. FUNDING: Swedish Research Council, Swedish Foundation for Strategic Research, Swedish Federal Government under the LUA/ALF agreement, Hoffmann La Roche, Cederoths, AstraZeneca, Sanofi-Aventis, Ethicon Endosurgery. PMID- 19556164 TI - Bariatric surgery, weight reduction, and cancer prevention. PMID- 19556166 TI - 2008 Congress of the ESFH in Dusseldorf, Germany. PMID- 19556165 TI - Suppression of IFNgamma+mycobacterial lipoarabinomannan-induced NO by IL-4 is due to decreased IRF-1 expression. AB - In mice, and possibly in humans, nitric oxide (NO) is an important host-defense molecule against Mycobacterium tuberculosis. Inducible nitric oxide synthase (iNOS) and NO are upregulated in murine macrophages stimulated with interferon gamma (IFNgamma) and mannose-capped lipoarabinomannan (ManLAM), a major lipoglycan in the cell wall of M. tuberculosis. Interleukin-4 (IL-4) can inhibit NO expression and may impair host immune response to M. tuberculosis. Therefore, we sought to determine the mechanism by which IL-4 inhibits IFNgamma+ManLAM induced NO production. Since l-arginine is the substrate for both iNOS and arginase, and IL-4 increases arginase activity by inducing its production, a plausible mechanism of IL-4 inhibition of NO expression is via depletion of l arginine through increased arginase activity. Herein, we show that IL-4 inhibited iNOS gene expression at the transcriptional level, suggesting an inhibitory mechanism that is independent of the competition for l-arginine between iNOS and arginase. Furthermore, pharmacologic inhibition of IL-4-induced arginase activity did not abrogate IL-4 inhibition of IFNgamma+ManLAM-induced NO expression. Instead, inhibition by IL-4 was mediated principally by the ability of IL-4 to inhibit the production of IFNgamma-induced interferon-gamma response factor-1 (IRF-1) protein, a critically important transcriptional element that enhances expression of IFNgamma-inducible genes such as iNOS. PMID- 19556167 TI - Lateralizing seizure focus in presurgical patients with temporal lobe epilepsy: utility of the Ruff-Light Trail Learning Test. AB - From a neuropsychological perspective, left or right temporal lobe abnormalities underlying seizures in patients with temporal lobe epilepsy (TLE) are thought to contribute to selective deficits in verbal or nonverbal memory abilities, respectively. The Ruff-Light Trail Learning Test (RULIT) is specifically designed to detect right hemisphere dysfunction. The purpose of our study was to examine the utility of the RULIT in distinguishing between patients with right (n=20) and patients with left (n=32) TLE in presurgical evaluations. We identified a significant between-group difference in RULIT scores, but not in the expected direction; that is, patients with right TLE had significantly better scores than patients with left TLE. These findings indicate that the RULIT may not be an appropriate test for presurgical epilepsy evaluations. Findings are discussed within the context of results from other lateralizing neuropsychological tests. PMID- 19556168 TI - Comparison of health care use and costs in newly diagnosed and established patients with fibromyalgia. AB - In 2004, the American Pain Society (APS) issued evidence-based fibromyalgia treatment recommendations. The objective of this claims database analysis is to describe prescription and medical use in patients with newly diagnosed and established fibromyalgia. Privately insured patients with 2+ myalgia/myositis claims (1999 to 2005) were categorized as newly diagnosed or established; this dichotomy involves comparisons between prediagnosis (S1) and postdiagnosis (S2) stages in the newly diagnosed and between newly diagnosed (S2) and established patients (S3). Use of APS guideline medications increased across stages: selective serotonin reuptake inhibitors (SSRIs) (S1, S2, S3: 20.6%, 22.9%, 25.3%), serotonin norepinephrine reuptake inhibitors (SNRIs) (4.5%, 6.4%, 8.9%), pregabalin/gabapentin (5.4%, 7.4%, 8.8%), benzodiazepines (19.0%, 21.1%, 24.2%), non-benzodiazepine sedatives (9.1%, 11.5%, 13.7%) (all P < .0001), and opioids (39.5%, 43.3%, 43.9%; S1 vs S2, P < .0001; S2 vs S3, P = .2835). Use of multiple therapeutic classes also increased across stages: 3+ classes (7.1%, 9.6%, 11.8%) (all P < .0001). Office visits to providers increased, on average, after diagnosis: primary care (70.9%, 78.3%, 76.3%; all P < .0001), chiropractors (28.8%, 51.1%, 53.3%; all P < .0001), rheumatologists (4.2%, 9.9%, 10.5%; S1 vs S2, P < .0001; S2 vs S3, P = .0595), mental health (6.4%, 7.3%, 8.3%; S1 vs S2, P < .0001, S2 vs S3, P = .0003). Average health care costs rose after diagnosis in the newly diagnosed group (S1: $6555 vs S2: $8654, P < .0001). PERSPECTIVE: This paper investigates prescription drug and medical care use with respect to stages of fibromyalgia diagnosis. Established fibromyalgia patients use more medical resources and have higher rates of concomitant medication use than newly diagnosed fibromyalgia patients. Findings can help educate providers regarding optimal drug treatment patterns in this population. PMID- 19556169 TI - Attachment relationships shape pain-signaling behavior. AB - Attachment relationships shape the manner in which children signal pain to others. Open communication of pain affect, inhibition of pain affect, and exaggeration of pain affect, reflect adaptations to different relationship contexts. The open and direct signaling of pain is adaptive in sensitive relationship contexts where caregivers respond to the distressed child with behaviors that facilitate protection, recovery, and healing. Inhibition of pain signals has survival advantages in situations where the open expressions of pain elicit negative parental responses (absence of caregiving, withdrawal from the child, or frank displeasure or anger). Exaggerated pain signaling functions as a means to elicit a caregiving response from preoccupied, inattentive, or neglectful attachment figures. This paper considers how a child's developmental experiences-specifically, the repeating person-specific experiences which make up attachment relationships-produce individual differences in the manner in which pain is experienced and signaled. PERSPECTIVE: This article reviews recent advances in our understanding of child development as articulated by contemporary attachment theory-in particular, the dynamic-maturational model (DMM)-and discusses their implications for interpreting human pain, pain-signaling behavior, and medically unexplained pain. The development of the experience of pain, along with ways of signaling pain, is tied to familial relationships generally and, in particular, to the manner in which attachment relationships shape the infant's behavior and physiology, thereby regulating the experience of pain. In explaining how the child's early attachment relationships produce individual differences in the way that she learns to experience and signal pain, the article provides an innovative perspective that is helpful in understanding the wide variations in patients' experience and presentation of pain, in elaborating formulations of medically unexplained pain, and in planning treatment. More broadly, the article suggests a variety of hypotheses for further empirical testing. PMID- 19556170 TI - Investigation of graft extenders in lumbar fusions: methodological and clinical concerns for current and future studies. PMID- 19556171 TI - Electro-acupuncture improves responsiveness to insulin via excitation of somatic afferent fibers in diabetic rats. AB - The effects of electro-acupuncture (EA) on plasma concentration of glucose and on responsiveness to insulin were examined in an animal model of diabetes, the streptozotocin-treated rat. Two weeks after treatment with streptozotocin, rats were anesthetized with urethane-chloralose and subjected to the EA for 10 min delivered to the tibialis anterior muscle of one side. The stimulation produced no significant changes in plasma glucose concentration. In contrast, EA increased the response of plasma glucose to insulin (0.2 U kg(-1)). The effect of EA on the responsiveness to insulin was abolished by section of both sciatic and femoral nerves ipsilateral to the side of the EA. These results show that EA in diabetic rats has no effect on plasma glucose concentration while it augments the responsiveness to insulin, and we show that this occurs via a mechanism that involves the somatic afferent nerves. PMID- 19556172 TI - Competition of copper and/or calcium in nickel-pectin interaction by potentiometric and voltammetric techniques. AB - The interaction between nickel and pectin extracted from citrus fruit was studied in 0.10 M KNO(3), at pH 5.5 and 25 degrees C. Differential pulse and/or square wave polarography were used to determine free nickel. For a high coverage degree (theta) of the pectin by the metal ion a good fitting was observed between experimental results and the model that includes both complex species, ML and ML(2) (M for the metal ion and L for the ligand). In the ML(2) species, Ni(II) interacts with two carboxylate groups of different chains, resulting in an inter chain association. For low theta values, the formation of ML(2) is hindered due to the repulsion between the negative charges of carboxylic groups in two independent segments of pectin. The influence of calcium or copper ions on the free nickel concentration, in the presence of pectin, may lead to a decrease in free nickel concentration, contrary to what would be expected from direct competition between Ca(II) or Cu(II) and Ni(II) for the pectin binding sites. This is due to the partial neutralisation of the negative carboxylic charges by the positive charges of the divalent cations, which favours NiL(2) formation through the association of independent chains. PMID- 19556173 TI - The North American Fetal Therapy Network (NAFTNet): a new approach to collaborative research in fetal diagnosis and therapy. AB - In August 2004, the National Institutes of Health organized a 'Workshop on Fetal Therapy' to develop a plan for the maternal-fetal, surgical, and neonatal evaluation and treatment of pregnancies that might benefit from in-utero therapy. At the completion of the workshop several recommendations were made, foremost of which was the 'formation of a cooperative group of clinical investigators to help set a national agenda for research and clinical progress in the field of fetal therapy'. Somewhat by coincidence, a multidisciplinary 'Fetal Therapy Working Group' that had been formed earlier in the year was well-positioned to accept this national mandate and proposed development of a North American Fetal Therapy Network (NAFTNet) to foster collaborative research between active fetal diagnosis and treatment centers in both the USA and Canada, develop a peer review mechanism for study proposals, explore ways to centralize data collection and study development, and establish an educational agenda for medical professionals and the public as well as training of future leaders in the field. NAFTNet represents a new paradigm and approach to international collaborative research. Early success has resulted in the recognition of the power of collaborative research efforts in studying rare congenital anomalies and intervention strategies to improve outcomes and survivals in such limited populations. By abandoning 'competitive research' for a cooperative, collaborative environment of research partnership, NAFTNet strives to be more responsible and effective in using limited resources and improving care for pregnancies and children born with congenital anomalies. PMID- 19556174 TI - 'Through-the-loop' tie for microsurgical suturing. AB - We present a technique for tying microsurgical knots called 'through-the-loop'. The main advantage of this technique is that the short end of the microsuture is always held by the instruments and thus prevented from falling and adhering to the surrounding tissue. Consequently, the short end can be easily re-grasped and the knot tied without difficulty. This technique can be performed by either right or left-hand-dominant surgeons and can facilitate faster and more efficient microsurgical knot tying, which the novice microsurgeon may find useful. PMID- 19556175 TI - Enhancing the viability of fat grafts using new transfer medium containing insulin and beta-fibroblast growth factor in autologous fat transplantation. AB - With the increasing application of liposuction in reconstructive surgery of the head and neck region and aesthetic plastic surgery of the facial region, autologous fat transplantation is widely used. However, due to the high absorption rate of fat grafts, a number of studies have examined the use of different agents to increase the viability of such grafts. Here, we evaluated the effects of a new transfer medium containing insulin and beta-fibroblast growth factor, which can increase the viability of fat grafts before performing autologous fat transplantation. Twenty-four New Zealand white rabbits were used in this study. For each rabbit, we harvested 4mg of fat from both inguinal regions. The samples from each rabbit were pooled, divided into four equal aliquots and then soaked in a transfer medium. Group I was the control group with normal saline as the transfer medium, group II used modified Dulbecco's modified Eagle's medium (modified DMEM), group III used modified DMEM with insulin and group IV used modified DMEM with insulin and beta-fibroblast growth factor. After dividing the dorsum of each rabbit into four even parts, one fat sample from each group was transplanted into one of the pockets created in the loose areolar tissue immediately beneath the dermis. Six rabbits were euthanised using pentobarbital sodium each time, 1, 3, 6 and 12 months after fat transplantation, and we measured the weights of the fat grafts and examined the histological changes using light microscopy. Groups III and IV showed significantly greater viability of fat grafts than the controls in the group I. On histological examination, large numbers of mature adipocytes were observed in groups III and IV, and reduced cyst formation and fibrosis were also observed. Autologous fat transplantation using the new transfer medium containing insulin and beta fibroblast growth factor is a simple, promising strategy for clinical application designed to augment the viability of fat transplantation. PMID- 19556178 TI - Use of coronary computed tomography angiography to detect coronary ostial stenosis after Bentall procedure. AB - Coronary ostial stenosis is a rare complication of the Bentall procedure for aortic root and aortic valve replacement. We report a case of coronary ostial stenosis after a Bentall procedure that was detected by coronary computed tomography angiography and subsequent percutaneous coronary interventions. PMID- 19556179 TI - Regional distribution of substance P within the amygdaloid complex and bed nucleus of the stria terminals. AB - The distribution of substance P (SP) within the amygdaloid complex and bed nucleus of the stria terminalis has been studied using a sensitive radioimmunoassay. Particularly high levels of SP were found in the medial nucleus of the amygdala. Large amounts of SP were also found in the various subdivisions of the bed nucleus. These results are in good agreement with immunohistochemical data. PMID- 19556177 TI - Noncardiac findings on cardiac CT part I: Pros and cons. AB - Cardiac computed tomography (CT) has evolved into an effective imaging technique for the evaluation of coronary artery disease in selected patients. Two distinct advantages over other noninvasive imaging modalities include its ability to evaluate directly the coronary arteries and to provide an opportunity to evaluate extracardiac structures, such as the lungs and mediastinum. Some centers reconstruct a small field of view (FOV) cropped around the heart, but a full FOV (from skin to skin in the irradiated area) is obtainable in the raw data of every scan so that clinically relevant noncardiac findings are identifiable. Debate in the scientific community has centered on the necessity for this large FOV evaluation. A review of noncardiac structures provides the opportunity to make alternative diagnoses that may account for the patient's presentation or to detect important but clinically silent problems such as lung cancer. Critics argue that the yield of biopsy-proven cancers is low and that the follow-up of incidental noncardiac findings is expensive, resulting in increased radiation exposure and possibly unnecessary further testing. In this two-part review we outline the issues surrounding the concept of the noncardiac read looking for noncardiac findings on cardiac CT. Part I focuses on the pros and cons of the practice of identifying noncardiac findings on cardiac CT. PMID- 19556180 TI - The effect of myelin basic protein (MBP) on the bioelectric activity of spinal cord and cerebellar neurones. AB - The effects of myelin basic protein (MBP) on the ventral root response in rat spinal cord in situ and on the spontaneous bioelectric activity of cultured rat Purkinje cells were investigated. Injection of 2 or 4 microl MBP (10(-4) M/l) into the motoneurone pool reduced the ventral root response to dorsal root stimulation. The depression was long-lasting (30-50 min) and partially reversible. At 5 x 10(-6) M/l MBP abolished the spontaneous activity of most cultured Purkinje cells, an effect which was not blocked by haloperidol, propranolol, phenoxybenzamine and bicuculline. The EC(50) for the depression was 4 + or - 1 x 10(-6) M/l. PMID- 19556181 TI - In vivo incorporation of exogenous stearic acid in synaptosomes: high occurrence of non-esterified fatty acids. AB - Subcutaneously injected stearic acid is taken up by the brain through the 'blood brain barrier' and is incorporated into synaptosomal lipids (as well as into other brain compartments). Phospholipids are potent acceptors (choline phosphatides, ethanolamine phosphatides, serine phosphatides). Moreover a high level of radioactivity was found in non-esterified fatty acids. PMID- 19556182 TI - Desensitization of glutamate responses - evidence for glutamate receptors in cat spinal cord. AB - Desensitization of spinal cord interneurons to prolonged application of glutamate was found to occur in 30% of the neurons tested. The response of the same neurons to aspartate was unaffected by prior desensitization to glutamate, suggesting the existence of separate receptors for these two amino acids. PMID- 19556183 TI - Regional effects of vasopressin on rat brain catecholamine metabolism. AB - Rats were injected i.c.v. with arginine(8)-vasopressin or saline 30 min after they had received an i.p. injection with the catecholamine synthesis inhibitor alpha-methyl-p-tyrosine. Three hours after the administration of vasopressin or saline the noradrenaline content of the hypothalamus, thalamus and medulla oblongata of the vasopressin-treated rats was significantly lower than that of the same parts of saline-treated rats. These results indicate that vasopressin increases nerve impulse flow in noradrenaline neurons in particular brain regions. PMID- 19556184 TI - Angiotensin-induced drinking in rats with hereditary hypothalamic diabetes insipidus. AB - Previous reports indicate that the pressor response seen following intracranially applied angiotensin II (AII) in normal rats is absent in hereditary hypothalamic diabetes insipidus (DI) animals. The dipsogenic effect of intracranially injected AII was therefore examined in DI rats and in control rats. Our results show that the drinking response to AII was present in DI animals and not different from control rats. Substitution of DI animals with vasopressin had no effect on the drinking response to AII. We conclude that it is possible to dissociate brain mechanisms mediating the centrally induced blood pressure response and the drinking response. PMID- 19556185 TI - Neurons in the human basolateral amygdala and hippocampal formation do not respond to odors. AB - Extracellular action potentials were recorded from human hippocampal, hippocampal gyrus, and basolateral amygdala neurons during passive and active olfactory stimulation. Introduction of an odor into a continuous stream of air passing over the olfactory mucosa resulted in no detectable change in firing. Actively sniffing from an odorous flask, whether or not the odor was detected or recognized, also failed to elicit an immediate unit response. However, after a latency of 10-15 sec, the slight hyperventilation inherent in sniffing did induce a strong change in firing by many neurons. In contrast to these unit responses, a short-latency EEG spindle was recorded in the amygdala that appeared directly related to mechanical stimulation of the olfactory mucosa. PMID- 19556186 TI - Interhemispheric field potentials, spreading depression and kindling. AB - The incidence of spreading depression, which was elicited by interhemispheric stimulation, increased with prolonged duration and strength of individual pulses and with longer stimulus trains. In the first trial spreading depression appeared in one-third of the experiments, in two-thirds it was activated by repeated stimulus trains. Both the activated and the first trial spreading depression were preceded by alternation of the interhemispheric response and often by self sustained afterdischarges. All the phases of the interhemispheric response patterns contained inhibitory components either in the form of phasic focal polarizing waves of the evoked and self-sustained field potentials or giant slow spreading depolarizations. The stimulation parameters determine whether a permanent increase of excitability prevails, as in Goddard's kindling procedures, or whether inhibitory patterns, described in this paper, result from the stimulation. PMID- 19556187 TI - Evidence for a non-dopaminergic action of amantadine. AB - Amantadine and dopamine have been applied by microiontophoresis to single neurones in the cerebral cortex. While the dopamine receptor blocker alpha flupenthixol antagonized responses to dopamine, it only partially reduced responses to amantadine. The effects of alpha-flupenthixol on dopamine and amantadine responses were highly significantly different (P < 0.001). Amantadine was probably not acting on noradrenaline receptors since it was unaffected by propranolol. Thus, amantadine may have a non-catecholaminergic action, which may explain the previously noted inability of catecholamine depletion to prevent the effects of the drug. PMID- 19556188 TI - Dimethoxyaporphine as an antagonist of chemical excitation in the pigeon optic tectum. AB - The alkaloid L-5,6-dimethoxyaporphine (L-nuciferine) was studied with iontophoretic techniques, and its interaction with excitant amino acids on neurones of the pigeon optic tectum was tested. This compound antagonizes the action of L-glutamate and, to a weaker extent, that of L-aspartate and d,L homocysteate. In comparison, L-5,6-dimethoxyaporphine did not reduce the acetylcholine evoked excitation. PMID- 19556189 TI - Behavioural stimulation induced by muscimol and other GABA agonists injected into the substantia nigra. AB - The unilateral injection of putative GABA agonists, muscimol, baclofen, imidazole acetic acid and GABA into the caudal area of substantia nigra (SN, zona reticulata) induced immediately a contralateral turning, whereas the antogonists picrotoxin and bicuculline methiodide induced ipsilateral turning. In the rostral area picrotoxin induced contralateral turning. Muscimol (10 ng bilaterally injected into SN) induced behavioural stimulation and antagonism of catelepsy in rats pretreated with high subcutaneous doses of perphenazine, haloperidol or reserpine and alpha-methyltyrosine. The muscimol behavioural stimulation seems dependent on a non-catecholaminergic neuronal pathway, probably present in the SN with GABA as the mediating neurotransmitter. PMID- 19556190 TI - Introduction to the special section on colloidal quantum dots for biomedical applications. PMID- 19556192 TI - A SVD-based method to assess the uniqueness and accuracy of SPECT geometrical calibration. AB - Geometrical calibration is critical to obtaining high resolution and artifact free reconstructed image for SPECT and CT systems. Most published calibration methods use analytical approach to determine the uniqueness condition for a specific calibration problem, and the calibration accuracy is often evaluated through empirical studies. In this work, we present a general method to assess the characteristics of both the uniqueness and the quantitative accuracy of the calibration. The method uses a singular value decomposition (SVD) based approach to analyze the Jacobian matrix from a least-square cost function for the calibration. With this method, the uniqueness of the calibration can be identified by assessing the nonsingularity of the Jacobian matrix, and the estimation accuracy of the calibration parameters can be quantified by analyzing the SVD components. A direct application of this method is that the efficacy of a calibration configuration can be quantitatively evaluated by choosing a figure-of merit, e.g., the minimum required number of projection samplings to achieve desired calibration accuracy. The proposed method was validated with a slit-slat SPECT system through numerical simulation studies and experimental measurements with point sources and an ultra-micro hot-rod phantom. The predicted calibration accuracy from the numerical studies was confirmed by the experimental point source calibrations at approximately 0.1 mm for both the center of rotation (COR) estimation of a rotation stage and the slit aperture position (SAP) estimation of a slit-slat collimator by an optimized system calibration protocol. The reconstructed images of a hot rod phantom showed satisfactory spatial resolution with a proper calibration and showed visible resolution degradation with artificially introduced 0.3 mm COR estimation error. The proposed method can be applied to other SPECT and CT imaging systems to analyze calibration method assessment and calibration protocol optimization. PMID- 19556191 TI - Segmentation of the outer vessel wall of the common carotid artery in CTA. AB - A novel method is presented for carotid artery vessel wall segmentation in computed tomography angiography (CTA) data. First the carotid lumen is semi automatically segmented using a level set approach initialized with three seed points. Subsequently, calcium regions located within the vessel wall are automatically detected and classified using multiple features in a GentleBoost framework. Calcium regions segmentation is used to improve localization of the outer vessel wall because it is an easier task than direct outer vessel wall segmentation. In a third step, pixels outside the lumen area are classified as vessel wall or background, using the same GentleBoost framework with a different set of image features. Finally, a 2-D ellipse shape deformable model is fitted to a cost image derived from both the calcium and vessel wall classifications. The method has been validated on a dataset of 60 CTA images. The experimental results show that the accuracy of the method is comparable to the interobserver variability. PMID- 19556193 TI - DT-REFinD: diffusion tensor registration with exact finite-strain differential. AB - In this paper, we propose the DT-REFinD algorithm for the diffeomorphic nonlinear registration of diffusion tensor images. Unlike scalar images, deforming tensor images requires choosing both a reorientation strategy and an interpolation scheme. Current diffusion tensor registration algorithms that use full tensor information face difficulties in computing the differential of the tensor reorientation strategy and consequently, these methods often approximate the gradient of the objective function. In the case of the finite-strain (FS) reorientation strategy, we borrow results from the pose estimation literature in computer vision to derive an analytical gradient of the registration objective function. By utilizing the closed-form gradient and the velocity field representation of one parameter subgroups of diffeomorphisms, the resulting registration algorithm is diffeomorphic and fast. We contrast the algorithm with a traditional FS alternative that ignores the reorientation in the gradient computation. We show that the exact gradient leads to significantly better registration at the cost of computation time. Independently of the choice of Euclidean or Log-Euclidean interpolation and sum of squared differences dissimilarity measure, the exact gradient achieves better alignment over an entire spectrum of deformation penalties. Alignment quality is assessed with a battery of metrics including tensor overlap, fractional anisotropy, inverse consistency and closeness to synthetic warps. The improvements persist even when a different reorientation scheme, preservation of principal directions, is used to apply the final deformations. PMID- 19556194 TI - Quantitative comparison of spot detection methods in fluorescence microscopy. AB - Quantitative analysis of biological image data generally involves the detection of many subresolution spots. Especially in live cell imaging, for which fluorescence microscopy is often used, the signal-to-noise ratio (SNR) can be extremely low, making automated spot detection a very challenging task. In the past, many methods have been proposed to perform this task, but a thorough quantitative evaluation and comparison of these methods is lacking in the literature. In this paper, we evaluate the performance of the most frequently used detection methods for this purpose. These include seven unsupervised and two supervised methods. We perform experiments on synthetic images of three different types, for which the ground truth was available, as well as on real image data sets acquired for two different biological studies, for which we obtained expert manual annotations to compare with. The results from both types of experiments suggest that for very low SNRs ( approximately 2), the supervised (machine learning) methods perform best overall. Of the unsupervised methods, the detectors based on the so-called h -dome transform from mathematical morphology or the multiscale variance-stabilizing transform perform comparably, and have the advantage that they do not require a cumbersome learning stage. At high SNRs ( > 5), the difference in performance of all considered detectors becomes negligible. PMID- 19556195 TI - Complex wavelet structural similarity: a new image similarity index. AB - We introduce a new measure of image similarity called the complex wavelet structural similarity (CW-SSIM) index and show its applicability as a general purpose image similarity index. The key idea behind CW-SSIM is that certain image distortions lead to consistent phase changes in the local wavelet coefficients, and that a consistent phase shift of the coefficients does not change the structural content of the image. By conducting four case studies, we have demonstrated the superiority of the CW-SSIM index against other indices (e.g., Dice, Hausdorff distance) commonly used for assessing the similarity of a given pair of images. In addition, we show that the CW-SSIM index has a number of advantages. It is robust to small rotations and translations. It provides useful comparisons even without a preprocessing image registration step, which is essential for other indices. Moreover, it is computationally less expensive. PMID- 19556196 TI - Noniterative MAP reconstruction using sparse matrix representations. AB - We present a method for noniterative maximum a posteriori (MAP) tomographic reconstruction which is based on the use of sparse matrix representations. Our approach is to precompute and store the inverse matrix required for MAP reconstruction. This approach has generally not been used in the past because the inverse matrix is typically large and fully populated (i.e., not sparse). In order to overcome this problem, we introduce two new ideas. The first idea is a novel theory for the lossy source coding of matrix transformations which we refer to as matrix source coding. This theory is based on a distortion metric that reflects the distortions produced in the final matrix-vector product, rather than the distortions in the coded matrix itself. The resulting algorithms are shown to require orthonormal transformations of both the measurement data and the matrix rows and columns before quantization and coding. The second idea is a method for efficiently storing and computing the required orthonormal transformations, which we call a sparse-matrix transform (SMT). The SMT is a generalization of the classical FFT in that it uses butterflies to compute an orthonormal transform; but unlike an FFT, the SMT uses the butterflies in an irregular pattern, and is numerically designed to best approximate the desired transforms. We demonstrate the potential of the noniterative MAP reconstruction with examples from optical tomography. The method requires offline computation to encode the inverse transform. However, once these offline computations are completed, the noniterative MAP algorithm is shown to reduce both storage and computation by well over two orders of magnitude, as compared to a linear iterative reconstruction methods. PMID- 19556197 TI - Color filter array demosaicking using high-order interpolation techniques with a weighted median filter for sharp color edge preservation. AB - Demosaicking is an estimation process to determine missing color values when a single-sensor digital camera is used for color image capture. In this paper, we propose a number of new methods based on the application of Taylor series and cubic spline interpolation for color filter array demosaicking. To avoid the blurring of an edge, interpolants are first estimated in four opposite directions so that no interpolation is carried out across an edge. A weighted median filter, whose filter coefficients are determined by a classifier based on an edge orientation map, is then used to produce an output from the four interpolants to preserve edges. Using the proposed methods, the original color can be faithfully reproduced with minimal amount of color artifacts even at edges. PMID- 19556198 TI - Hierarchical ensemble of global and local classifiers for face recognition. AB - In the literature of psychophysics and neurophysiology, many studies have shown that both global and local features are crucial for face representation and recognition. This paper proposes a novel face recognition method which exploits both global and local discriminative features. In this method, global features are extracted from the whole face images by keeping the low-frequency coefficients of Fourier transform, which we believe encodes the holistic facial information, such as facial contour. For local feature extraction, Gabor wavelets are exploited considering their biological relevance. After that, Fisher's linear discriminant (FLD) is separately applied to the global Fourier features and each local patch of Gabor features. Thus, multiple FLD classifiers are obtained, each embodying different facial evidences for face recognition. Finally, all these classifiers are combined to form a hierarchical ensemble classifier. We evaluate the proposed method using two large-scale face databases: FERET and FRGC version 2.0. Experiments show that the results of our method are impressively better than the best known results with the same evaluation protocol. PMID- 19556199 TI - Improved dot diffusion by diffused matrix and class matrix co-optimization. AB - Dot diffusion is an efficient approach which utilizes concepts of block-wise and parallel-oriented processing to generate halftones. However, the block-wise nature of processing reduces image quality much more significantly as compared to error diffusion. In this work, four types of filters with various sizes are employed in co-optimization procedures with class matrices of size 8 n 8 and 16 x 16 to improve the image quality. The optimal diffused weighting and area are determined through simulations. Many well-known halftoning methods, some of which includes direct binary search (DBS), error diffusion, ordered dithering, and prior dot diffusion methods, are also included for comparisons. Experimental results show that the proposed dot diffusion achieved quality close to some forms of error diffusion, and additionally, superior to the well-known Jarvis and Stucki error diffusion and Mese's dot diffusion. Moreover, the inherent parallel processing advantage of dot diffusion is preserved, allowing us to reap higher executing efficiency than both DBS and error diffusion. PMID- 19556200 TI - An edge-weighted centroidal Voronoi tessellation model for image segmentation. AB - Centroidal Voronoi tessellations (CVTs) are special Voronoi tessellations whose generators are also the centers of mass (centroids) of the Voronoi regions with respect to a given density function and CVT-based methodologies have been proven to be very useful in many diverse applications in science and engineering. In the context of image processing and its simplest form, CVT-based algorithms reduce to the well-known k -means clustering and are easy to implement. In this paper, we develop an edge-weighted centroidal Voronoi tessellation (EWCVT) model for image segmentation and propose some efficient algorithms for its construction. Our EWCVT model can overcome some deficiencies possessed by the basic CVT model; in particular, the new model appropriately combines the image intensity information together with the length of cluster boundaries, and can handle very sophisticated situations. We demonstrate through extensive examples the efficiency, effectiveness, robustness, and flexibility of the proposed method. PMID- 19556201 TI - Cyclorotation models for eyes and cameras. AB - The human visual system obeys Listing's law, which means that the cyclorotation of the eye (around the line of sight) can be predicted from the direction of the fixation point. It is shown here that Listing's law can conveniently be formulated in terms of rotation matrices. The function that defines the observed cyclorotation is derived in this representation. Two polynomial approximations of the function are developed, and the accuracy of each model is evaluated by numerical integration over a range of gaze directions. The error of the simplest approximation for typical eye movements is less than half a degree. It is shown that, given a set of calibrated images, the effect of Listing's law can be simulated in a way that is physically consistent with the original camera. This condition is important for robotic models of human vision, which typically do not reproduce the mechanics of the oculomotor system. PMID- 19556202 TI - A note on two methods for estimating missing pairwise preference values. AB - This note analyzes two methods for calculating missing values of an incomplete reciprocal fuzzy preference relation. The first method by Herrera-Viedma appeared in the IEEE Transactions on Systems, Man, and Cybernetics-Part B: Cybernetics [vol. 37, no. 1 (2007) 176-189], while the second one by Fedrizzi and Giove appeared later in the European Journal of Operational Research [vol. 183 (2007) 303-313]. The underlying concept driving both methods is the additive consistency property. We show that both methods, although different, are very similar. Both methods derive the same estimated values for the independent-missing-comparison case, while they differ in the dependent-missing-comparison case. However, it is shown that a modification of the first method coincides with the second one. Regarding the total reconstruction of an incomplete preference relation, it is true that the second method performs worse than the first one. When Herrera Viedma 's method is unsuccessful, Fedrizzi-Giove's method is as well. However, in those cases when Fedrizzi-Giove's method cannot guarantee the successful reconstruction of an incomplete preference relation, we have that Herrera-Viedma 's method can. These results lead us to claim that both methods should be seen as complementary rather than competitors in their application, and as such, we propose a reconstruction policy of incomplete fuzzy preference relations using both methods. By doing this, the only unsuccessful reconstruction case is when there is a chain of missing pairwise comparisons involving each one of the feasible alternatives at least once. PMID- 19556203 TI - EvoOligo: oligonucleotide probe design with multiobjective evolutionary algorithms. AB - Probe design is one of the most important tasks in successful deoxyribonucleic acid microarray experiments. We propose a multiobjective evolutionary optimization method for oligonucleotide probe design based on the multiobjective nature of the probe design problem. The proposed multiobjective evolutionary approach has several distinguished features, compared with previous methods. First, the evolutionary approach can find better probe sets than existing simple filtering methods with fixed threshold values. Second, the multiobjective approach can easily incorporate the user's custom criteria or change the existing criteria. Third, our approach tries to optimize the combination of probes for the given set of genes, in contrast to other tools that independently search each gene for qualifying probes. Lastly, the multiobjective optimization method provides various sets of probe combinations, among which the user can choose, depending on the target application. The proposed method is implemented as a platform called EvoOligo and is available for service on the web. We test the performance of EvoOligo by designing probe sets for 19 types of Human Papillomavirus and 52 genes in the Arabidopsis Calmodulin multigene family. The design results from EvoOligo are proven to be superior to those from well-known existing probe design tools, such as OligoArray and OligoWiz. PMID- 19556204 TI - Attitude adaptation in satisficing games. AB - Satisficing game theory offers an alternative to classical game theory that describes a flexible model of players' social interactions. Players' utility functions depend on other players' attitudes rather than simply their actions. However, satisficing players with conflicting attitudes may enact dysfunctional behaviors, which results in poor performance. We present an evolutionary method by which a population of players may adapt their attitudes to improve payoff. In addition, we extend the Nash-equilibrium concept to satisficing games, showing that the method leads players toward the equilibrium in their attitudes. We apply these ideas to the stag hunt--a simple game in which cooperation does not easily evolve from noncooperation. The evolutionary method provides two major contributions. First, satisficing players may improve their performance by adapting their attitudes. Second, numerical results demonstrate that cooperation in the stag hunt can emerge much more readily under the method we present than under traditional evolutionary models. PMID- 19556205 TI - Computational intelligence in bioinformatics: SNP/haplotype data in genetic association study for common diseases. AB - Comprehensive evaluation of common genetic variations through association of single-nucleotide polymorphism (SNP) structure with common complex disease in the genome-wide scale is currently a hot area in human genome research due to the recent development of the Human Genome Project and HapMap Project. Computational science, which includes computational intelligence (CI), has recently become the third method of scientific enquiry besides theory and experimentation. There have been fast growing interests in developing and applying CI in disease mapping using SNP and haplotype data. Some of the recent studies have demonstrated the promise and importance of CI for common complex diseases in genomic association study using SNP/haplotype data, especially for tackling challenges, such as gene gene and gene-environment interactions, and the notorious "curse of dimensionality" problem. This review provides coverage of recent developments of CI approaches for complex diseases in genetic association study with SNP/haplotype data. PMID- 19556206 TI - Eigenleads: ECG leads for maximizing information capture and improving SNR. AB - There is currently much interest in exploring new ways to optimize ECG acquisition. In the current study, we have investigated optimal configurations of ECG leads with respect to: 1) best signal magnitude (maximal signal variance) and 2) best reconstruction of the total body surface potential distribution and the 12-lead ECG. Principal component analysis was applied to a set of 117-lead body surface potential maps (BSPMs) recorded from 559 subjects. Three bipolar leads, referred to as "eigenleads," were identified from the extrema on the resulting eigenvectors. Recording sites for the three leads were largely located in the precordial region. The magnitude of the signals recorded from the eigenleads was calculated on a set of 185 unseen subjects. The accuracy of the eigenleads in the reconstruction of BSPMs and the 12-lead ECG was also assessed for each subject. These results were compared to existing limited lead systems. It was found that, when compared to conventional leads, eigenleads could be used to increase signal strength (rms voltage) by 27.9%, 39.0%, and 20.3% for P-waves, QRS segments, and STT segments, respectively. Although the eigenleads were not able to reconstruct total body surface information as well as the 12-lead ECG (24.4 mu V versus 20.2 mu V), the eigenleads did perform comparably with other limited lead systems in the estimation of the 12-lead ECG. In particular, the eigenleads performed well in the reconstruction of precordial leads in comparison to the EASI lead system and a limited lead system made up of a subset of precordial leads. The proposed leads are a suitable alternative limited leads system, and can be used to improve SNR. More work is needed to test the practicality of such leads. PMID- 19556209 TI - Ichthyosis congenita, harlequin fetus type: a case report. AB - Ichthyosis is a very heterogeneous family of skin disorders with harlequin ichthyosis being the most severe genetic form. It is a rare autosomal recessive condition, characterized by dry, severely thickened skin with large plates of hyperkeratotic scale, separated by deep fissures. Infants are very susceptible to metabolic abnormalities and infections. They usually do not survive for very long, but several long term survivals have been noted. The vast majority of affected individuals are homozygous for mutations in the ABCA12 gene, which cause a deficiency of the epidermal lipid transporter, resulting in hyperkeratosis and abnormal barrier function. We report a case of a newborn with harlequin ichthyosis, born to unrelated parents, who had a favorable evolution with topical treatment and intensive care. PMID- 19556210 TI - Sodium-dependent phosphate cotransporter type 1 sequence polymorphisms in male patients with gout. AB - OBJECTIVES: Molecular biological approaches have recently identified urate transporters in renal proximal tubular cells. Human sodium-dependent phosphate cotransporter type 1 encoded by SLC17A1 is a urate transporter localised to the renal proximal tubular cells and candidate molecule to secret urate from renal tubular cells to urine. This study investigated the roles of SLC17A1 in the development of gout. PATIENTS AND METHODS: Single nucleotide polymorphisms in the human SLC17A1 gene (rs1165176, rs1165151, rs1165153, rs1165196, rs1165209, rs1165215, rs1179086, rs3799344 and rs3757131) were selected, and an association study was conducted using male patients with gout (n=175) and male controls (n=595). RESULTS: There were significant differences between gout and control groups in the distribution of genotypes at rs1165196 (T806C; Ile269Thr, odds ratio (OR) 0.55, p=0.0035), rs1179086 (OR 0.57, p=0.0018) and rs3757131 (OR 0.54, p=0.0026). In controls, T806C alone had no effect on serum uric acid (sUA) levels. However, T806C showed significant interaction with a reduction of sUA in obese individuals (body mass index > or = 25) using multiple regression analysis. CONCLUSIONS: Our data suggest that SLC17A1 polymorphisms are associated with the development of gout. PMID- 19556211 TI - Loss of imprinting of IGF2 characterises high IGF2 mRNA-expressing type of fibroblast-like synoviocytes in rheumatoid arthritis. AB - OBJECTIVE: Increased expression of insulin-like growth factor 2 (IGF2) by fibroblast-like synoviocytes (FLS) was associated with low inflammatory synovium of patients with rheumatoid arthritis (RA). The aim of this study was to analyse whether the differential expression of IGF2, whose expression is normally restricted to one allele, is due to activation of the normally suppressed allele. METHODS: IGF2 gene expression of RA FLS was quantified by quantitative real-time PCR. FLS heterozygous for a 3'-untranslated region IGF2 polymorphism were selected to measure the relative contribution of the allelic transcripts by allele-specific transcript quantification assay. Proliferation was determined by [(3)H]thymidine incorporation. RESULTS: IGF2 was shown to contribute to RA FLS proliferation. FLS could be classified in IGF2 high and IGF2 low-expressing cell lines. Allelic IGF2 transcript quantification analysis revealed that in part of the RA FLS the normally suppressed allele was activated, resulting in biallelic expression of the IGF2 gene. Biallelic expression was associated with increased levels of IGF2 mRNA production. CONCLUSION: The findings indicate that the imprinting status of IGF2 might underlie the increased expression of IGF2, which may contribute to autonomous growth of RA FLS of low inflammatory synovial tissues. PMID- 19556212 TI - Predictive factors of visual and anatomical outcome after intravitreal bevacizumab treatment of neovascular age-related macular degeneration: an optical coherence tomography study. AB - AIM: To evaluate the baseline visual and optical coherence tomography (OCT) factors on outcomes after intravitreal bevacizumab treatment of subfoveal neovascular age-related macular degeneration (AMD). METHODS: A retrospective analysis of 73 eyes treated with intravitreal bevacizumab for subfoveal neovascular AMD was performed. Change in best corrected Snellen visual acuity (BCVA) and central retinal thickness (CRT) on OCT were the primary outcomes. Automated and manual measurements were made for all OCT characteristics. RESULTS: Seventy-three (100%) and 58 (79.5%) eyes were followed for 3 and 6 months, respectively. The mean BCVA improved from 20/177 to 20/160 (p = 0.03) at 3 months and to 20/143 (p = 0.04) at 6 months. The mean CRT decreased 93 microm (p<0.0001) and 105 microm (p<0.0001) at 3 and 6 months, respectively. Baseline BCVA worse than 20/100 was associated with greater visual improvement (p< or =0.04). Eyes with baseline CRT greater than 400 microm experienced a greater mean CRT reduction (p<0.05). Treatment-naive patients had a greater mean CRT reduction than those previously treated with any modality (p<0.05) CONCLUSIONS: Baseline BCVA and CRT positively influence mean visual and CRT improvement, respectively, after intravitreal bevacizumab in wet AMD. Any prior treatment predicted less CRT reduction. PMID- 19556213 TI - Major shifts in corneal transplantation procedures in north China: 5316 eyes over 12 years. AB - AIMS: To investigate the major shifts in the ratio of lamellar keratoplasty (LKP) to penetrating keratoplasty (PKP) and in the preoperative indications for each procedure. METHODS: Medical records of patients who received LKP and/or PKP at Shandong Eye Institute between 1996 and 2007 were categorised and reviewed. The time period was divided into intervals of 1996-8, 1999-2001, 2002-4 and 2005-7. RESULTS: A total of 4346 patients (5316 eyes) with integrated clinical records were included in the study. LKPs and PKPs were performed on 1558 eyes (29.3%) and 3758 eyes (70.7%), respectively. Within the first 3-year interval, the top three indications for LKP were chemical burns, keratoconus and corneal dermoid; the top reasons for PKP were viral keratitis, suppurative keratitis and corneal scarring. Within the last interval, suppurative keratitis, keratoconus and viral keratitis became most common indications for LKP and suppurative keratitis, viral keratitis and bullous keratopathy for PKP. The ratio of LKP to PKP operations tended to increase. CONCLUSION: Following proper indications, the use of LKP has increased in number in north China and has become particularly frequent in the management of corneal infections, keratoconus, corneal degeneration, and stromal dystrophy. PMID- 19556214 TI - Anti-vascular endothelial growth factor agents for diabetic maculopathy. AB - The management of diabetic macular oedema is changing. The therapeutic armamentarium for diabetic macular oedema (DMO) includes a new group of drugs that inhibit vascular endothelial growth factor (VEGF). These anti-VEGF agents are already being used widely in DMO in clinical practice despite that several phase III trials on these drugs are still underway. There are no established protocols on the use of these agents in DMO, but short-term results are appealing. This review provides an update on the use of anti-VEGF agents in DMO. Although intravitreal delivery of anti-VEGF agents is a relatively safe procedure, the long-term local and systemic effects of these agents in the diabetic population remain unknown. In this regard, this review also highlights the need for close surveillance of the use of these drugs in this high-risk population. PMID- 19556215 TI - The genetics of central corneal thickness. AB - Evidence in the recent literature has highlighted the importance of central corneal thickness (CCT) in relation to several ocular and non-ocular conditions. Most notably, thinner CCT has been identified as a risk factor for open-angle glaucoma. Despite having an extensive knowledge of the structure and function of the cornea, little is known about the pathways that determine CCT. There are data to suggest however that CCT has a strong genetic component. Heritability studies conducted in twins and family pedigrees indicate that CCT is one of the most highly heritable human traits, whereas data from a diverse range of ethnic groups show clear ethnic-related differences in CCT. Extreme CCT measurements have also been associated with rare genetic diseases. Although there is strong evidence supporting a genetic component to normal CCT variation, to date, no genes have been identified. This review investigates the current literature surrounding this topic and explores the significance of understanding the genetics of CCT and how this might benefit the field of open-angle glaucoma treatment and research. PMID- 19556216 TI - A steroid-inducible promoter for the cornea. AB - AIM: Topical glucocorticosteroids are administered to virtually every corneal transplant recipient, but irreversible immunological rejection remains the leading cause of graft failure. Ex vivo gene therapy of the donor cornea has been shown to modulate graft rejection in experimental models. The efficacy of a glucocorticosteroid-inducible promoter was assessed in controlling transgene expression following lentivirus-mediated gene transfer to ovine and human corneas. METHODS: A glucocorticosteroid response element (GRE5) was cloned into a lentiviral vector (LV-GRE-IL10) encoding the model transgene interleukin 10. Transgene expression by LV-GRE-IL10-transduced A549 cells, ovine corneas and human corneas cultured with or without dexamethasone was quantified by an IL10 specific enzyme-linked immunosorbent assay. RESULTS: IL10 levels were 30-40-fold higher in supernatants from LV-GRE-IL10-transduced A549 cells cultured with dexamethasone than in controls. Dexamethasone withdrawal resulted in restoration of baseline IL10 levels. Supernatants from LV-GRE-IL10-transduced ovine and human corneas cultured in dexamethasone contained nine to 10 times more IL10 than supernatants from transduced corneas cultured without dexamethasone. CONCLUSION: The GRE5 promoter in a lentiviral vector drove rapid, sustained and inducible transgene expression in both ovine and human corneas in the presence of dexamethasone. A steroid-inducible promoter may be useful for controlling transgene expression in gene-modified donor corneal allografts. PMID- 19556217 TI - Metastatic retinoblastoma: single institution experience over two decades. AB - BACKGROUND: Metastatic spread in retinoblastoma is a rare occurrence in developed countries but still associated with a poor prognosis. PATIENTS AND METHODS: Medical records of all metastatic retinoblastoma diagnosed during a 20-year period were retrospectively reviewed. RESULTS: Six patients out of 104 presented a metastatic disease with an incidence at diagnosis of 2%. Three had a metastatic disease at diagnosis, one patient a trilateral retinoblastoma and two a metastatic spread after enucleation. All but one were sporadic retinoblastoma. Central nervous system (CNS) involvement was reported in five patients, while one patient had an intraorbital lesion, and bone and bone marrow spread. Different treatment strategies were administered based on local treatment plus chemotherapy and radiotherapy with or without high-dose chemotherapy. An ifosfamide/carboplatin/etoposide regimen was administered in three patients resulting in a partial response. Out of six patients, four died, and two patients are alive at 60 and 63 months from diagnosis. Both children with a long follow-up were treated with high-dose chemotherapy. All but one of the patients with CNS involvement died; the survivor was a patient with pineal involvement. CONCLUSION: This retrospective review confirms a curable strategy based on local treatment and conventional plus high-dose chemotherapy. Patients with CNS involvement remain incurable. PMID- 19556218 TI - Trends in overweight and obesity prevalence in Dutch, Turkish, Moroccan and Surinamese South Asian children in the Netherlands. AB - OBJECTIVE: To determine trends in the prevalence of overweight and obesity in children in The Hague (the Netherlands) from 1999 through 2007. DESIGN: Population-based study of a series of cross-sectional assessments of height and weight from electronic health records. SETTING: Child Health Care (Municipal Health Service), The Hague. PARTICIPANTS: 50,961 children aged 3-16 years, with Dutch (59%), Turkish (17%), Moroccan (13%) or Surinamese South Asian (11%) ethnicity, representative of the four major ethnic groups in The Hague, with 85,234 weight and height measurements recorded in 1999-2007. MAIN OUTCOME MEASURES: (Trends in) the prevalence of overweight (excluding obesity) and obesity as defined by the International Obesity Taskforce cut-off points, using logistic regression with year as independent variable. RESULTS: From 1999 through 2007 there was a decrease in the prevalence of overweight in Dutch girls from 12.6% to 10.9% (OR 0.96; 95% CI 0.95 to 0.98) and an increase in Turkish boys from 14.6% to 21.4% (OR 1.08; 95% CI 1.04 to 1.11). Obesity prevalence rose significantly in Turkish boys from 7.9% to 13.1% (OR 1.04; 95% CI 1.01 to 1.06) and in Turkish girls from 8.0% to 10.7% (OR 1.04; 95% CI 1.01 to 1.08). Dutch boys, and Moroccan and Surinamese South Asian boys and girls showed no significant trends. CONCLUSIONS: The declining prevalence of overweight in Dutch girls may indicate reversal of previous trends in the Netherlands. However, in Turkish children overweight prevalence and obesity is high and increasing. Further public health action is necessary, especially for Turkish children. PMID- 19556219 TI - A protective effect of breastfeeding on the progression of non-alcoholic fatty liver disease. AB - OBJECTIVE: Non-alcoholic fatty liver disease (NAFLD) is a spectrum of liver disease characterised by accumulation of large-droplet fat in hepatocytes with possible progression to inflammation and fibrosis. Breastfeeding has benefits for child health, both during infancy and later in life, reducing the risk of manifestations of the metabolic syndrome. Here we investigated the association between early type of feeding (breastfed versus formula-fed and duration of breastfeeding) and later NAFLD development. STUDY DESIGN: We investigated 191 young Caucasian children (3-18 years old) with NAFLD consecutively enrolled between January 2003 and September 2007 in our centre. 48% of these children (n = 91) had been breastfed for a median (interquartile range) time of 8 (7) months. RESULTS: After correction for age, waist circumference, gestational age and neonatal weight, the odds of non-alcoholic steatohepatitis (NASH) (OR 0.04, 95% CI 0.01 to 0.10) and fibrosis (OR 0.32, 95% CI 0.16 to 0.65) were lower in breastfed versus not breastfed infants. Moreover, the odds of NASH (OR 0.70, exact 95% CI 0.001 to 0.87) and fibrosis (OR 0.86, exact 95% CI 0.75 to 0.98) decreased for every month of breastfeeding. CONCLUSIONS: This observational study suggests that earlier feeding habits might affect the clinical expression of NASH from 3 to 18 years later, with an apparent drug-like preventive effect of breastfeeding. PMID- 19556220 TI - Cerebral venous sinus thrombosis: a case series including thrombolysis. AB - BACKGROUND: Cerebral venous sinus thrombosis (CVST) in children is associated with a high incidence of serious morbidity and mortality. The presenting features are variable. It can be diagnostically challenging and the optimal treatment is uncertain. AIM: To describe the features of a series of children with CVST treated in a single paediatric neurology centre and to discuss the role of local thrombolysis. METHODS: Electronic databases were searched using diagnostic labels and International Classification of Diseases (ICD) codes to identify children aged 1 month to under 17 years with CVST. Their records were reviewed. RESULTS: 21 children were identified over a period of 8.25 years with a median age of 7.1 years. The presenting symptoms included headache (15 children), vomiting (14 children) and visual disturbance (eight children). Signs found included papilloedema (16 children), fever (six children) and sixth nerve palsy (six children). The most common underlying condition was middle ear infection (13 children). All cases received unfractionated heparin and four severe cases received local pharmacological thrombolysis. 48% of cases had an adverse outcome (death, chronic intracranial hypertension, residual hemiparesis or sixth nerve palsy). DISCUSSION: CVST has non-specific presenting features and a high risk of significant morbidity. CVST is typically found in association with a predisposing condition. Although heparin is the mainstay of treatment, thrombolysis may reverse deterioration as seen in three cases in this series. However, there is insufficient evidence to recommend the routine use of thrombolysis at present. PMID- 19556221 TI - RNA interference targeted to the conserved dimerization initiation site (DIS) of HIV-1 restricts virus escape mutation. AB - Short hairpin RNAs (shRNA) targeting viral or cellular genes can effectively inhibit human immunodeficiency virus type 1 (HIV-1) replication. This inhibition, however, may induce mutations in the targeted gene, leading to rapid escape from the shRNA-induced inhibition. We generated a lymphoid cell line that stably expressed a 19-bp shRNA targeting a well-conserved dimerization initiation site (DIS) of HIV-1, which strongly inhibited viral replication, thereby delaying virus escape. Furthermore, treatment of HIV-1 infection with DIS- and vif-shRNA combination therapy resulted in superior anti-viral responses compared to vif shRNA monotherapy. Continuous challenge with HIV-1, however, generated virus mutants that could overcome the RNA interference restriction. Such anti-genes may be promising tools for HIV-1 gene therapy for HIV/acquired immunodeficiency syndrome. PMID- 19556223 TI - Overexpression of Plk3 causes morphological change and cell growth suppression in Ras pathway-activated cells. AB - To unravel the growth inhibition mechanism of Polo-like kinase 3 (Plk3), the effect of overexpression of Plk3 was examined in 293T cells. Cell rounding, changes in actin organization and cellular detachment were induced by Plk3 transfection in a kinase activity-dependent manner. Although apoptosis was not observed, Plk3 overexpression suppressed cellular growth in a long-term colony forming assay. Because both Plk3 and Ras affect F-actin organization, the effect of co-transfection of Plk3 and Ras was evaluated. Adhesion was synergistically lost by co-transfection of these two genes, compared with transfection of Plk3 alone. Furthermore, overexpression of Plk3 caused long-term growth suppression in Ras-transformed NIH3T3. Collectively, Plk3 activation might cause cytoskeleton re organization and result in growth suppression more pronouncedly in Ras pathway activated cells. PMID- 19556222 TI - Storage of gangliosides GM2 and fucosyl GM1 in the kidney of MCC strain of mastomys (Praomys coucha). AB - Previously, we histochemically examined the kidney of the MCC strain of mastomys (Praomys coucha) and found the storage of gangliosides. In the present studies, the lipid-bound sialic acid content of gangliosides in the MCC kidney was about 9 to 14-fold higher than that of the control (MWC strain). In the MCC kidney, sialic acids of male gangliosides were composed of N-acetylneuraminic acid at 91.5%; sialic acids of female gangliosides, however, were composed almost entirely of N-glycolylneuraminic acid. TLC of gangliosides showed that the MCC kidney contained four abundant gangliosides (two gangliosides each in males and females). These gangliosides isolated by HPLC were identified to be GM2(NeuAc) and fucosyl GM1(NeuAc) in the male MCC kidney and GM2(NeuGc) and fucosyl GM1(NeuGc) in the female MCC kidney by secondary ion mass spectrometry, TLC/immunostaining and TLC after enzyme treatments. Although the MCC kidney contained control levels of the activities of beta-N-acetylhexosaminidase, alpha l-fucosidase, N-acetylgalactosaminyltransferase and fucosyltransferase, the activity of beta-galactosidase in the MCC kidney was increased to 400-500% of that in the MWC kidney. Therefore, we discussed the possibility that in the MCC kidney, GM2 was abundantly produced by the effect of increased beta-galactosidase activity. PMID- 19556224 TI - Changes in the conformation of the Vsr endonuclease amino-terminal domain accompany DNA cleavage. AB - In Escherichia coli, T/G mismatches arising from deamination of 5-methylcytosine to thymine are converted to CG base pairs by the very short patch (VSP) repair pathway. DNA Polymerase I removes and resynthesizes the mismatched T starting from a 5'-nick created by the Vsr endonuclease. We used limited trypsinolysis to probe conformational changes in the N-terminal domain of Vsr in response to DNA binding, DNA cleavage and interaction with the polymerase. Our data show that the domain becomes trypsin resistant only under conditions that allow DNA cleavage, while interaction with the polymerase restores trypsin sensitivity. We suggest that the domain changes its conformation as a result of DNA nicking, and that DNA Pol I releases Vsr from the nick by reversing that conformational change. PMID- 19556225 TI - Insights into the enzyme-substrate interaction in the norovirus 3C-like protease. AB - The Glu54 residue of the norovirus 3C-like protease was implicated in proteolysis as a third-member carboxylate of the catalytic triad. The E54L mutant protease cleaved the sequence (133)LSFE/AP between the 3B and 3C regions of norovirus polyprotein, but did not cleave the sequence (198)ATSE/GK between the 3A and 3B. The 3BC junction mutation (3B-L133A or 3B-F135S) hampered the cleavage by the E54L protease, whereas the 3AB junction mutation (3A-A198L, S200F) allowed the E54L protease to digest. These results indicate that the E54L mutant protease is a substrate-specificity mutant and requires large hydrophobic amino acid residues at both P4 and P2 positions of the substrate. It was notable that the 3A-S200F P2 position mutation caused tight interaction between the wild-type protease and the C-terminus of the 3A protein, hence a decreased release rate of the product from the enzyme. This tight binding was dependent on the hydrophobicity of amino acid residues introduced at position 200 of the 3A region and was affected by the mutation in the bII-cII loop of the protease or the mutation of position 198 of 3A corresponding to the P4 position of the substrate. These results suggest that the protease and the substrate sense each other in the process of the proteolysis, being supported by crystal structures. PMID- 19556226 TI - Thoracic outlet syndrome associated with a large cervical rib. AB - This case illustrates the anatomic impact of a large cervical rib as an etiology for thoracic outlet syndrome. Current management remains predicated on astute diagnosis, multidisciplinary therapy including, physical therapy, scalene muscle blockade, and surgical decompression in appropriate cases. PMID- 19556227 TI - Basic science review. Vascular distensibility as a predictive tool in the management of small asymptomatic abdominal aortic aneurysms. AB - BACKGROUND: This study investigates whether baseline aortic wall distensibility serves as a supplemental bio-marker for AAA progression and need for later repair. METHODS: In 1998, 61 males with a small asymptomatic AAAs had a baseline measurement of elasticity and stiffness, using an echo-tracking ultrasound system (Diamove). The cohort was followed till 2005 concerning Dmax, expansion rate, operations for AAA, hospitalisation do to cardiovascular disease and death. RESULTS: During follow-up, 49% died, and 45.9% were hospitalised do to cardiovascular disease, compared to Dmax, Ep and b no significant associations were found. Elasticity correlated moderately to annual expansion rate and Dmax. Good correlation was found between annual expansion rate and Dmax. ROC-curve analysis showed that elasticity, stiffness and Dmax all tended to predict future need for AAA-repair. CONCLUSION: Baseline aortic wall distensibility may provide an additional parameter for AAA to optimize the indication and time for elective repair. PMID- 19556228 TI - Is infrainguinal percutaneous atherectomy better suited for certain arteries than others? AB - OBJECTIVE: We analyzed our results with percutaneous rotational atherectomy catheters and specifically examined whether they were more likely to be associated with a successful outcome when used to treat smaller diameter vessels such as infrapopliteal (IP) arteries compared to larger diameter femoropopliteal (FP) arteries and infrainguinal arterial autogenous vein grafts (GRAFTS). MATERIAL AND METHODS: Between January 1, 2005, and December 31, 2006, athrectomies were performed on 32 patients for claudication (14), gangrene (9), rest pain (4), and failing GRAFTS (5). Treated vessels included 14 superficial femoral, 1 popliteal, 5 anterior tibial, 4 posterior tibial, and 3 peroneal arteries along with 5 failing GRAFTS. All procedures were performed by vascular surgeons in an endovascular operating suite using a mobile C-arm. Results for larger diameter vessels including FP arteries and GRAFTS were combined (FP + BYPASSES) and compared to results of IP artery lesions. Follow-up averaged 10 weeks (range, 0.5-34 weeks). RESULTS: Length of treated lesions averaged 4.2 cm (range, 1-15 cm) for FP + GRAFT lesions (9 occlusions, 11 stenoses) versus 1.8 cm (1-4 cm) for IP lesions (6 occlusions, 6 stenoses; P = ns). Procedural success rate based on postoperative segmental pressures, pulse volume recordings, and duplex ultrasound was 70% (14/20) for FP + GRAFTS versus 83% (10/12) for IP lesions (P = .03). Need for concomitant adjunctive balloon angioplasty to treat residual stenosis tended to be higher for FP + GRAFT lesions (40% [8/20]) compared to IP lesions (25% [3/12]; P = ns). During follow-up, 25% (5/20) of FP + GRAFTS lesions required reintervention (3 balloon angioplasties, 1 thrombectomy, 1 GRAFT pseudoaneurysm) versus none for the IP lesions (P = .03). Limb salvage rates were 90% (18/20) for FP + GRAFT lesions versus 100% (12/12) for IP lesions during this short follow-up. CONCLUSIONS: These preliminary results suggest that short segment IP arterial stenoses and occlusions can be successfully treated with atherectomy catheters with a lower rate of reintervention during short-term follow-up, less need for concomitant adjunctive balloon angioplasty and a lower complication rate compared to FP + graft lesions. PMID- 19556229 TI - Newton's Law to the rescue: therapeutic effects of gravity aiding the management of a migratory venous foreign body--a case report. AB - The management of intravascular metallic foreign bodies (FB) can be difficult and challenging. We report a case of a migrating FB, initially within the femoral vein which subsequently migrated to the intrahepatic vena-cava. Following a change of posture, the metallic FB moved with gravity against the normal venous blood flow to the left renal vein. It was finally fixed in position in a peripheral branch of the renal vein using an intravascular stent. Employing gravity as a therapeutic intervention and the technique used in isolating the FB has not, to our knowledge, been reported before. A case is described, and the literature is reviewed. PMID- 19556230 TI - Inflammatory abdominal aortic aneurysms (IAAA): past and present. AB - AIM: The aim of the study is to determine whether presentation and outcomes of inflammatory abdominal aortic aneurysms (IAAA) have changed over the last five decades. METHODS: Comparison of current outcomes (January 2001 to December 2007) with results of the earliest report from our unit in 1972. RESULTS: In contemporary series, 421 patients underwent AAA repair; 38 (9%) were IAAA. In 58% patients, IAAA was an incidental finding, whereas 42% patients were symptomatic with abdominal or back pain. Of those, 32% were ruptured IAAA. Male-to-female ratio was 12:1. Thirty-day mortality was 13%; elective 11.5%; emergency 17%. Comparison with 1972 study showed no change in the incidence and gender predilection. Presentation as an incidental finding and rupture increased 4- and 2-folds, respectively. CONCLUSION: The incidence and gender predilection of IAAA have remained unchanged. The 4-fold increase in the presentation as an incidental finding reflects current trends in patient evaluation. PMID- 19556231 TI - Is balloon angioplasty of peri-anastomotic stenoses of failing peripheral arterial bypasses worthwhile? AB - INTRODUCTION: Surgical revision of failing peripheral arterial bypass grafts has generally been shown to provide superior patency rates compared to balloon angioplasty. We analyzed whether balloon angioplasty, specifically of peri anastomotic stenoses (PAS), provided acceptable patency rates, because surgery for these lesions is more difficult and is likely associated with higher complication rates compared to surgical revision of stenoses in the body of a graft. METHODS: This is a retrospective review of PAS balloon angioplasties performed at a single institution between January 1, 1999, and September 1, 2005. We report ''primary site patency'' as a stenosis treated by balloon angioplasty, ''revised primary site patency'' as a stenosis treated by repeat balloon angioplasty, and ''secondary site patency'' as an angioplastied stenosis treated surgically or when the graft thrombosed and was revised surgically. All procedures were performed in an endovascular operating room based on duplex scan findings suggesting a significant stenosis. RESULTS: 48 PAS in 33 autologous vein and 15 prosthetic grafts were treated by balloon angioplasty in 42 patients. Mean follow-up was 12 months (range, 1-49 months). Interventions were performed on 22 femoropopliteal grafts (11 proximal, 11 distal), 20 femorotibial grafts (5 proximal, 15 distal), 2 axillofemoral grafts (2 proximal anastomoses), 2 popliteal-pedal grafts (1 proximal, 1 distal), and 1 common iliac-femoral graft (proximal). Life-table analysis revealed 2-year primary, assisted primary, and secondary patency rates of 38%, 58%, and 84%, respectively. No major complications occurred with any endovascular intervention. CONCLUSION: Balloon angioplasty of PAS resulted in acceptable 2-year assisted primary patency rate of almost 60%. Endovascular intervention avoided repeat incisions in scarred groins, higher rates of nerve injury and infection, significant blood loss, and longer length of hospital stays. We recommend that balloon angioplasty of PAS be attempted before resorting to surgical intervention, especially in cases of hostile anastomotic wounds. PMID- 19556232 TI - Effect of hospital volume on in-hospital mortality for renal artery bypass. AB - BACKGROUND: A recent report determined that the nationwide mortality for renal artery bypass (RAB) is surprisingly high-10%. We hypothesized that operative mortality for RAB is related to the volume of such operations performed in each center. METHODS: The Nationwide Inpatient Sample was analyzed to identify patients undergoing RAB for the years 2000-2005. In-hospital mortality for RAB was compared between hospitals. RESULTS: During the study period, RAB was performed on 7413 patients with an overall in-hospital mortality of 9.6%. The multivariate logistic regression analyses revealed that after adjusting for surgical risk, increasing hospital volume was significantly associated with decreased in-hospital mortality for RAB (odds ratio 0.98; 95% confidence interval, 0.96-0.99; P=.015). CONCLUSIONS: Patient risk profile and hospital volume are critical determinants of in-hospital mortality for RAB, which should be factored into decision making for patients requiring intervention for renovascular disease. PMID- 19556233 TI - Architectural plasticity in young Eucalyptus marginata on restored bauxite mines and adjacent natural forest in south-western Australia. AB - The aboveground architecture of Eucalyptus marginata (Jarrah) was investigated in chronosequences of young trees (2.5, 5 and 10 m height) growing in a seasonally dry climate in a natural forest environment with intact soils, and on adjacent restored bauxite mine sites on soils with highly modified A and B horizons above an intact C horizon. Compared to forest trees, trees on restored sites were much younger and faster growing, with straighter, more clearly defined main stems and deeper, narrower crowns containing a greater number of branches that were longer, thinner and more vertically angled. Trees on restored sites also had a higher fraction of biomass in leaves than forest trees, as indicated by 20-25% thicker leaves, 30-70% greater leaf area, 10-30% greater leaf area to sapwood area ratios and 5-30% lesser branch Huber values. Differences in crown architecture and biomass distribution were consistent with putatively greater soil-water, nutrient and light availability on restored sites. Our results demonstrate that under the same climatic conditions, E. marginata displays a high degree of plasticity of aboveground architecture in response to the net effects of resource availability and soil environment. These differences in architecture are likely to have functional consequences in relation to tree hydraulics and growth that, on larger scales, is likely to affect the water and carbon balances of restored forest ecosystems. This study highlights substrate as a significant determinant of tree architecture in water-limited environments. It further suggests that the architecture of young trees on restored sites may need to change again if they are to survive likely longer-term changes in resource availability. PMID- 19556234 TI - Carbohydrate storage in wood and bark of rubber trees submitted to different level of C demand induced by latex tapping. AB - When the current level of carbohydrates produced by photosynthesis is not enough to meet the C demand for maintenance, growth or metabolism, trees use stored carbohydrates. In rubber trees (Hevea brasiliensis Muell. Arg.), however, a previous study (Silpi U., A. Lacointe, P. Kasemsap, S. Thanisawanyangkura, P. Chantuma, E. Gohet, N. Musigamart, A. Clement, T. Ameglio and P. Thaler. 2007. Carbohydrate reserves as a competing sink: evidence from tapping the rubber tree. Tree Physiol. 27:881-889) showed that the additional sink created by latex tapping results not in a decrease, but in an increase in the non-structural carbohydrate (NSC) storage in trunk wood. In this study, the response of NSC storage to latex tapping was further investigated to better understand the trade off between latex regeneration, biomass and storage. Three tapping systems were compared to the untapped Control for 2 years. Soluble sugars and starch were analyzed in bark and wood on both sides of the trunk, from 50 to 200 cm from the ground. The results confirmed over the 2 years that tapped trees stored more NSC, mainly starch, than untapped Control. Moreover, a double cut alternative tapping system, which produced a higher latex yield than conventional systems, led to even higher NSC concentrations. In all tapped trees, the increase in storage occurred together with a reduction in trunk radial growth. This was interpreted as a shift in carbon allocation toward the creation of reserves, at the expense of growth, to cover the increased risk induced by tapping (repeated wounding and loss of C in latex). Starch was lower in bark than in wood, whereas it was the contrary for soluble sugars. The resulting NSC was twice as low and less variable in bark than in wood. Although latex regeneration occurs in the bark, changes related to latex tapping were more marked in wood than in bark. From seasonal dynamics and differences between the two sides of the trunk in response to tapping, we concluded that starch in wood behaved as the long-term reserve compartment at the whole trunk level, whereas starch in bark was a local buffer. Soluble sugars behaved like an intermediate, ready-to-use compartment in both wood and bark. Finally, the dynamics of carbohydrate reserves appears a relevant parameter to assess the long-term performance of latex tapping systems. PMID- 19556235 TI - Woody-to-total area ratio determination with a multispectral canopy imager. AB - Leaf area index (LAI) - defined as one half of the total green leaf area per unit ground surface area - can be determined by direct or indirect methods. Three major sources of errors exist in indirect LAI measurements: within-shoot clumping, beyond-shoot clumping and non-photosynthetic components. The effect of non-photosynthetic components on LAI measurements can be described by the woody to-total area ratio, alpha; however, no convenient and efficient indirect methods have been developed to estimate alpha, especially the variations in alpha with zenith angle , alpha(theta). We describe the development and use of a multispectral canopy imager (MCI) to estimate alpha and alpha(theta) by considering the effects of non-random distributions of canopy elements and woody components and the overestimation of needle-to-shoot area ratio on woody components. The MCI, which mainly comprises a near-infrared band camera (Fujifilm IS-1), two visible band cameras (Canon 40D), filters and a pan tilt, was developed to measure clumping index, woody-to-total area ratio and geometric parameters of isolated trees. Two typical sampling plots (Plots 1 and 5) chosen from among 16 permanent forest experiment plots were selected for the estimation of alpha and alpha(theta). The non-random distributions of canopy elements and woody components were estimated separately at eight zenith angles (from 0 degrees to 70 degrees in increments of 10 degrees) using MCI images based on the gap size distribution theory. The visible/near-infrared image pairs captured by the MCI were able to discriminate among sky, leaves, cloud and woody components. Based on three methods of estimation, we obtained woody-to-total area ratios of 0.24, 0.19, 0.19 for Plot 1 and 0.23, 0.18, 0.17 for Plot 5. If clumping effects were ignored, alpha values were overestimated by as much as 21% and 24% at Plots 1 and 5, respectively. We demonstrated that alpha(theta) varied with the zenith angle, with variations in the range of 3-33% at Plot 1 and 2-65% at Plot 5. A new formula for the precise determination of LAI is proposed. PMID- 19556236 TI - Oligomerization is crucial for the stability and function of heme oxygenase-1 in the endoplasmic reticulum. AB - Heme oxygenase-1 (HO-1), a stress-inducible enzyme anchored in the endoplasmic reticulum (ER) by a single transmembrane segment (TMS) located at the C terminus, interacts with NADPH cytochrome P450 reductase and biliverdin reductase to catalyze heme degradation to biliverdin and its metabolite, bilirubin. Previous studies suggested that HO-1 functions as a monomer. Using chemical cross-linking, co-immunoprecipitation, and fluorescence resonance energy transfer (FRET) experiments, here we showed that HO-1 forms dimers/oligomers in the ER. However, oligomerization was not observed with a truncated HO-1 lacking the C-terminal TMS (amino acids 266-285), which exhibited cytosolic and nuclear localization, indicating that the TMS is essential for the self-assembly of HO-1 in the ER. To identify the interface involved in the TMS-TMS interaction, residue Trp-270, predicted by molecular modeling as a potential interfacial residue of TMS alpha helices, was mutated, and the effects on protein subcellular localization and activity assessed. The results showed that the W270A mutant was present exclusively in the ER and formed oligomers with similar activity to those of the wild type HO-1. Interestingly, the W270N mutant was localized not only in the ER, but also in the cytosol and nucleus, suggesting it is susceptible to proteolytic cleavage. Moreover, the microsomal HO activity of the W270N mutant was significantly lower than that of the wild type. The W270N mutation appears to interfere with the oligomeric state, as revealed by a lower FRET efficiency. Collectively, these data suggest that oligomerization, driven by TMS-TMS interactions, is crucial for the stabilization and function of HO-1 in the ER. PMID- 19556237 TI - Cadmium induces retinoic acid signaling by regulating retinoic acid metabolic gene expression. AB - The transition metal cadmium is an environmental teratogen. In addition, cadmium and retinoic acid can act synergistically to induce forelimb malformations. The molecular mechanism underlying the teratogenicity of cadmium and the synergistic effect with retinoic acid has not been addressed. An evolutionarily conserved gene, beta,beta-carotene 15,15'-monooxygenase (BCMO), which is involved in retinoic acid biosynthesis, was studied in both Caenorhabditis elegans and murine Hepa 1-6 cells. In C. elegans, bcmo-1 was expressed in the intestine and was cadmium inducible. Similarly, in Hepa 1-6 cells, Bcmo1 was induced by cadmium. Retinoic acid-mediated signaling increased after 24-h exposures to 5 and 10 microm cadmium in Hepa 1-6 cells. Examination of gene expression demonstrated that the induction of retinoic acid signaling by cadmium may be mediated by overexpression of Bcmo1. Furthermore, cadmium inhibited the expression of Cyp26a1 and Cyp26b1, which are involved in retinoic acid degradation. These results indicate that cadmium-induced teratogenicity may be due to the ability of the metal to increase the levels of retinoic acid by disrupting the expression of retinoic acid-metabolizing genes. PMID- 19556238 TI - Localizing the membrane binding region of Group VIA Ca2+-independent phospholipase A2 using peptide amide hydrogen/deuterium exchange mass spectrometry. AB - The Group VIA-2 Ca(2+)-independent phospholipase A(2) (GVIA-2 iPLA(2)) is composed of seven consecutive N-terminal ankyrin repeats, a linker region, and a C-terminal phospholipase catalytic domain. No structural information exists for this enzyme, and no information is known about the membrane binding surface. We carried out deuterium exchange experiments with the GVIA-2 iPLA(2) in the presence of both phospholipid substrate and the covalent inhibitor methyl arachidonoyl fluorophosphonate and located regions in the protein that change upon lipid binding. No changes were seen in the presence of only methyl arachidonoyl fluorophosphonate. The region with the greatest change upon lipid binding was region 708-730, which showed a >70% decrease in deuteration levels at numerous time points. No decreases in exchange due to phospholipid binding were seen in the ankyrin repeat domain of the protein. To locate regions with changes in exchange on the enzyme, we constructed a computational homology model based on homologous structures. This model was validated by comparing the deuterium exchange results with the predicted structure. Our model combined with the deuterium exchange results in the presence of lipid substrate have allowed us to propose the first structural model of GVIA-2 iPLA(2) as well as the interfacial lipid binding region. PMID- 19556239 TI - PR55 alpha, a regulatory subunit of PP2A, specifically regulates PP2A-mediated beta-catenin dephosphorylation. AB - A central question in Wnt signaling is the regulation of beta-catenin phosphorylation and degradation. Multiple kinases, including CKI alpha and GSK3, are involved in beta-catenin phosphorylation. Protein phosphatases such as PP2A and PP1 have been implicated in the regulation of beta-catenin. However, which phosphatase dephosphorylates beta-catenin in vivo and how the specificity of beta catenin dephosphorylation is regulated are not clear. In this study, we show that PP2A regulates beta-catenin phosphorylation and degradation in vivo. We demonstrate that PP2A is required for Wnt/beta-catenin signaling in Drosophila. Moreover, we have identified PR55 alpha as the regulatory subunit of PP2A that controls beta-catenin phosphorylation and degradation. PR55 alpha, but not the catalytic subunit, PP2Ac, directly interacts with beta-catenin. RNA interference knockdown of PR55 alpha elevates beta-catenin phosphorylation and decreases Wnt signaling, whereas overexpressing PR55 alpha enhances Wnt signaling. Taken together, our results suggest that PR55 alpha specifically regulates PP2A mediated beta-catenin dephosphorylation and plays an essential role in Wnt signaling. PMID- 19556241 TI - A map of Drosophila melanogaster small nuclear RNA-activating protein complex (DmSNAPc) domains involved in subunit assembly and DNA binding. AB - Transcription of genes coding for the small nuclear RNAs (snRNAs) is dependent upon a unique transcription factor known as the small nuclear RNA-activating protein complex (SNAPc). SNAPc binds to an essential proximal sequence element located about 40-65 base pairs upstream of the snRNA transcription start site. In the fruit fly Drosophila melanogaster, DmSNAPc contains three distinct polypeptides (DmSNAP190, DmSNAP50, and DmSNAP43) that are stably associated with each other and bind to the DNA as a complex. We have used mutational analysis to identify domains within each subunit that are involved in complex formation with the other two subunits in vivo. We have also identified domains in each subunit required for sequence-specific DNA binding. With one exception, domains required for subunit-subunit interactions lie in the most evolutionarily conserved regions of the proteins. However, DNA binding by DmSNAPc is dependent not only upon the conserved regions but is also highly dependent upon domains outside the conserved regions. Comparison with functional domains identified in human SNAPc indicates many parallels but also reveals significant differences in this ancient yet rapidly evolving system. PMID- 19556240 TI - The hedgehog pathway transcription factor GLI1 promotes malignant behavior of cancer cells by up-regulating osteopontin. AB - The role of Hedgehog (Hh) signaling as a developmental pathway is well established. Several recent studies have implicated a role for this pathway in multiple cancers. In this study we report that expression of GLI1 and osteopontin (OPN) increase progressively with the progression of melanoma from primary cutaneous cancer to metastatic melanoma in clinically derived specimens. We have further determined that OPN is a direct transcriptional target of GLI1. We have observed that OPN expression is stimulated in the presence of Hh ligands and inhibited in the presence of the Smoothened (SMO) inhibitor, cyclopamine. Transcriptional silencing of GLI1 negatively impacts OPN expression and compromises the ability of cancer cells to proliferate, migrate, and invade in vitro and interferes with their ability to grow as xenografts and spontaneously metastasize in nude mice. These altered attributes could be rescued by re expressing OPN in the GLI1-silenced cells, suggesting that OPN is a critical downstream effector of active GLI1 signaling. Our observations lead us to conclude that the GLI1-mediated up-regulation of OPN promotes malignant behavior of cancer cells. PMID- 19556242 TI - Transforming growth factor-beta (TGF-beta1) activates TAK1 via TAB1-mediated autophosphorylation, independent of TGF-beta receptor kinase activity in mesangial cells. AB - Transforming growth factor-beta1 (TGF-beta1) is a multifunctional cytokine that signals through the interaction of type I (TbetaRI) and type II (TbetaRII) receptors to activate distinct intracellular pathways. TAK1 is a serine/threonine kinase that is rapidly activated by TGF-beta1. However, the molecular mechanism of TAK1 activation is incompletely understood. Here, we propose a mechanism whereby TAK1 is activated by TGF-beta1 in primary mouse mesangial cells. Under unstimulated conditions, endogenous TAK1 is stably associated with TbetaRI. TGF beta1 stimulation causes rapid dissociation from the receptor and induces TAK1 phosphorylation. Deletion mutant analysis indicates that the juxtamembrane region including the GS domain of TbetaRI is crucial for its interaction with TAK1. Both TbetaRI-mediated TAK1 phosphorylation and TGF-beta1-induced TAK1 phosphorylation do not require kinase activity of TbetaRI. Moreover, TbetaRI-mediated TAK1 phosphorylation correlates with the degree of its association with TbetaRI and requires kinase activity of TAK1. TAB1 does not interact with TGF-beta receptors, but TAB1 is indispensable for TGF-beta1-induced TAK1 activation. We also show that TRAF6 and TAB2 are required for the interaction of TAK1 with TbetaRI and TGF beta1-induced TAK1 activation in mouse mesangial cells. Taken together, our data indicate that TGF-beta1-induced interaction of TbetaRI and TbetaRII triggers dissociation of TAK1 from TbetaRI, and subsequently TAK1 is phosphorylated through TAB1-mediated autophosphorylation and not by the receptor kinase activity of TbetaRI. PMID- 19556243 TI - Cold shock domain protein 3 regulates freezing tolerance in Arabidopsis thaliana. AB - In response to cold, Escherichia coli produces cold shock proteins (CSPs) that have essential roles in cold adaptation as RNA chaperones. Here, we demonstrate that Arabidopsis cold shock domain protein 3 (AtCSP3), which shares a cold shock domain with bacterial CSPs, is involved in the acquisition of freezing tolerance in plants. AtCSP3 complemented a cold-sensitive phenotype of the E. coli CSP quadruple mutant and displayed nucleic acid duplex melting activity, suggesting that AtCSP3 also functions as an RNA chaperone. Promoter-GUS transgenic plants revealed tissue-specific expression of AtCSP3 in shoot and root apical regions. When exposed to low temperature, GUS activity was extensively induced in a broader region of the roots. In transgenic plants expressing an AtCSP3-GFP fusion, GFP signals were detected in both the nucleus and cytoplasm. An AtCSP3 knock-out mutant (atcsp3-2) was sensitive to freezing compared with wild-type plants under non-acclimated and cold-acclimated conditions, whereas expression of C-repeat-binding factors and their downstream genes during cold acclimation was not altered in the atcsp3-2 mutant. Overexpression of AtCSP3 in transgenic plants conferred enhanced freezing tolerance over wild-type plants. Together, the data demonstrated an essential role of RNA chaperones for cold adaptation in higher plants. PMID- 19556244 TI - Association of cell surface mucins with galectin-3 contributes to the ocular surface epithelial barrier. AB - Maintenance of an intact mucosal barrier is critical to preventing damage to and infection of wet-surfaced epithelia. The mechanism of defense has been the subject of much investigation, and there is evidence now implicating O glycosylated mucins on the epithelial cell surface. Here we investigate a new role for the carbohydrate-binding protein galectin-3 in stabilizing mucosal barriers through its interaction with mucins on the apical glycocalyx. Using the surface of the eye as a model system, we found that galectin-3 colocalized with two distinct membrane-associated mucins, MUC1 and MUC16, on the apical surface of epithelial cells and that both mucins bound to galectin-3 affinity columns in a galactose-dependent manner. Abrogation of the mucin-galectin interaction in four different mucosal epithelial cell types using competitive carbohydrate inhibitors of galectin binding, beta-lactose and modified citrus pectin, resulted in decreased levels of galectin-3 on the cell surface with concomitant loss of barrier function, as indicated by increased permeability to rose bengal diagnostic dye. Similarly, down-regulation of mucin O-glycosylation using a stable tetracycline-inducible RNA interfering system to knockdown c1galt1 (T synthase), a critical galactosyltransferase required for the synthesis of core 1 O-glycans, resulted in decreased cell surface O-glycosylation, reduced cell surface galectin-3, and increased epithelial permeability. Taken together, these results suggest that galectin-3 plays a key role in maintaining mucosal barrier function through carbohydrate-dependent interactions with cell surface mucins. PMID- 19556245 TI - On the mechanism of multiple lysine methylation by the human mixed lineage leukemia protein-1 (MLL1) core complex. AB - Transcription in eukaryotic genomes depends on enzymes that regulate the degree of histone H3 lysine 4 (H3K4) methylation. The mixed lineage leukemia protein-1 (MLL1) is a member of the SET1 family of H3K4 methyltransferases and is frequently rearranged in acute leukemias. Despite sequence comparisons that predict that SET1 family enzymes should only monomethylate their substrates, mono , di-, and trimethylation of H3K4 has been attributed to SET1 family complexes in vivo and in vitro. To better understand this paradox, we have biochemically reconstituted and characterized a five-component 200-kDa MLL1 core complex containing human MLL1, WDR5, RbBP5, Ash2L, and DPY-30. We demonstrate that the isolated MLL1 SET domain is a slow monomethyltransferase and that tyrosine 3942 of MLL1 prevents di- and trimethylation of H3K4. In contrast, a complex containing the MLL1 SET domain, WDR5, RbBP5, Ash2L, and DPY-30, displays a marked approximately 600-fold increase in enzymatic activity but only to the dimethyl form of H3K4. Single turnover kinetic experiments reveal that the reaction leading to H3K4 dimethylation involves the transient accumulation of a monomethylated species, suggesting that the MLL1 core complex uses a non processive mechanism to catalyze multiple lysine methylation. We have also discovered that the non-SET domain components of the MLL1 core complex possess a previously unrecognized methyltransferase activity that catalyzes H3K4 dimethylation within the MLL1 core complex. Our results suggest that the mechanism of multiple lysine methylation by the MLL1 core complex involves the sequential addition of two methyl groups at two distinct active sites within the complex. PMID- 19556246 TI - Reduced expression of an RNA-binding protein by prolactin leads to translational silencing of programmed cell death protein 4 and apoptosis in newt spermatogonia. AB - Recent studies indicate that the balance between cell survival and proapoptotic signals determines which cells commit to life or death. We have shown that the balance between follicle-stimulating hormone and prolactin determines differentiation or apoptosis in 7th generation spermatogonia during newt spermatogenesis; however, the molecular mechanisms specifying their fate are poorly understood. Here we show that the newt RNA-binding protein (nRBP) plays a critical role in determining their fate. nRBP was identified as a clone whose mRNA is decreased by prolactin, resulting in the reduction of the protein, which is otherwise expressed predominantly in the spermatogonia. nRBP protein associated with the mRNA for newt programmed cell death protein 4 (nPdcd4) at the 3'-untranslated region. nRBP reduction increased nPdcd4 mRNA but decreased its protein. In a cell-free system, cytoplasmic extracts containing reduced amounts of nRBP and nPdcd4 protein induced apoptosis, whereas adding nRBP protein to the extracts blocked apoptosis. Furthermore, overexpression of nRBP protected cells from apoptosis, stabilized the chimeric transcript containing the nPdcd4 3' untranslated region, and accelerated its translation. These data suggest that, in the absence of nRBP, nPdcd4 mRNA is not stabilized and its translation is suppressed, leading to apoptosis in the spermatogonia. PMID- 19556248 TI - The policies and politics of creating a comparative clinical effectiveness research center. AB - As part of the early efforts of the Obama administration to begin health care reform, $1.1 billion for comparative effectiveness research was included in the stimulus bill. Although this amount can be considered as an initial down payment, difficult issues such as where to place an ongoing effort, the role of such research in informing clinical decision making or reducing health care spending, and the governance to ensure full involvement by stakeholders have not yet been resolved. Legislation proposed over the past two years offers some insights into the options available going forward. PMID- 19556247 TI - Novel roles of GATA1 in regulation of angiogenic factor AGGF1 and endothelial cell function. AB - AGGF1 is an angiogenic factor, and its deregulation is associated with a vascular malformation consistent with Klippel-Trenaunay syndrome (KTS). This study defines the molecular mechanism for transcriptional regulation of AGGF1 expression. Transcription of AGGF1 starts at two nearby sites, -367 and -364 bp upstream of the translation start site. Analyses of 5'- and 3'-serial promoter deletions defined the core promoter/regulatory elements, including two repressor sites (from -1971 to -3990 and from -7521 to -8391, respectively) and two activator sites (a GATA1 consensus binding site from -295 to -300 and a second activator site from -129 to -159). Both the GATA1 site and the second activator site are essential for AGGF1 expression. A similar expression profile was found for GATA1 and AGGF1 in cells (including various endothelial cells) and tissues. Electrophoretic mobility shift assay and chromatin immunoprecipitation assays demonstrated that GATA1 was able to bind to the AGGF1 DNA in vitro and in vivo. Overexpression of GATA1 increased expression of AGGF1. We identified one rare polymorphism -294C>T in a sporadic KTS patient, which is located in the GATA1 site, disrupts binding of GATA1 to DNA, and abolishes the GATA1 stimulatory effect on transcription of AGGF1. Knockdown of GATA1 expression by siRNA reduced expression of AGGF1, and resulted in endothelial cell apoptosis and inhibition of endothelial capillary vessel formation and cell migration, which was rescued by purified recombinant human AGGF1 protein. These results demonstrate that GATA1 regulates expression of AGGF1 and reveal a novel role for GATA1 in endothelial cell biology and angiogenesis. PMID- 19556249 TI - 'Marginal medicine': targeting comparative effectiveness research to reduce waste. AB - Many policymakers believe that comparative effectiveness research will reduce "waste" in the U.S. health care system by providing guidance on what health care options work best for different patients. How such research will produce findings that identify waste has not been clearly elucidated. This paper presents a conceptual framework for different evidentiary categories of what we call "marginal medicine." This framework could help clinicians, researchers, and policymakers align the results of comparative effectiveness research with efforts to identify opportunities to improve the value of health care services. PMID- 19556250 TI - Long-term follow-up free of ventricular fibrillation recurrence after resuscitated cardiac arrest in a myotonic dystrophy type 1 patient. AB - Cardiac involvement in myotonic dystrophy type 1 (DM1) is frequent with increased incidence of conduction disturbances and sudden cardiac death when compared with general population. We describe a 38-year-old man in whom the diagnosis of DM1 was made 8 years after occurrence of cardiac arrest owing to ventricular fibrillation and discuss management of DM1 patients at risk for sudden cardiac death. PMID- 19556251 TI - Sinus rhythm restoration by catheter ablation in patients with long-lasting atrial fibrillation and congestive heart failure: impact of the left ventricular ejection fraction improvement on the implantable cardioverter defibrillator insertion indication. AB - AIMS: In the setting of congestive heart failure (CHF), atrial fibrillation (AF) ablation can improve clinical status and the left ventricular ejection fraction (LVEF) value. However, the impact of AF ablation on the implantable cardioverter defibrillator (ICD) indication has never been specifically addressed. METHODS AND RESULTS: Study subject were six CHF (mean age 61.1 +/- 6.9 years, mean LVEF 25.8 +/- 7.3%) patients refractory to conventional medical treatment with long-lasting AF unresponsive to external cardioversion. Five patients had an idiopathic dilated cardiomyopathy (DCM) and one had an ischaemic cardiomyopathy (ICM). Their New York Heart Association (NYHA) class was III-IV. Two patients had renal insufficiency. No patient had left ventricular delay. All patients underwent AF ablation. LVEF and NYHA class were dramatically improved in the five DCM patients. New York Heart Association class, but not the LVEF, was improved in the ICM patient. A redo ablative procedure was undertaken in four of five DCM patients and in the ICM patient due to arrhythmia recurrence. Left ventricular ejection fraction and NYHA were improved again in the DCM patients (56 +/- 4.4%, I-II, respectively) and led to ICD indication preclusion. The LVEF remained low in the ICM patient (30%) and led to ICD insertion. Sinus rhythm has been stable during the 18.1 +/- 5.7 months follow-up period. CONCLUSION: Atrial fibrillation ablation in CHF patients can improve both the clinical status of patients and their LVEF, especially among those affected by DCM. The LVEF improvement has the potential to preclude the indication for a primary prevention ICD insertion. PMID- 19556252 TI - Unexpected asystole during 3T magnetic resonance imaging of a pacemaker-dependent patient with a 'modern' pacemaker. AB - Pacemaker patients might safely undergo magnetic resonance imaging (MRI) if several prudent suggestions are followed by the supervising physician. Controversy exists regarding the safety of scanning all device patients, but especially pacemaker-dependent patients. Despite following suggestions previously proposed, physicians should be prepared for unexpected potentially life threatening events during the conduct of MRI. PMID- 19556253 TI - Deregulation of EIF4E: a novel mechanism for autism. AB - BACKGROUND: Autism is a common childhood onset neurodevelopmental disorder, characterised by severe and sustained impairment of social interaction and social communication, as well as a notably restricted repertoire of activities and interests. Its aetiology is multifactorial with a strong genetic basis. EIF4E is the rate limiting component of eukaryotic translation initiation, and plays a key role in learning and memory through its control of translation within the synapse. EIF4E mediated translation is the final common process modulated by the mammalian target of rapamycin (mTOR), PTEN and fragile X mental retardation protein (FMRP) pathways, which are implicated in autism. Linkage of autism to the EIF4E region on chromosome 4q has been found in genome wide linkage studies. METHODS AND RESULTS: The authors present evidence that directly implicates EIF4E in autism. In a boy with classic autism, the authors observed a de novo chromosome translocation between 4q and 5q and mapped the breakpoint site to within a proposed alternative transcript of EIF4E. They then screened 120 autism families for mutations and found two unrelated families where in each case both autistic siblings and one of the parents harboured the same single nucleotide insertion at position -25 in the basal element of the EIF4E promoter. Electrophoretic mobility shift assays and reporter gene studies show that this mutation enhances binding of a nuclear factor and EIF4E promoter activity. CONCLUSIONS: These observations implicate EIF4E, and more specifically control of EIF4E activity, directly in autism. The findings raise the exciting possibility that pharmacological manipulation of EIF4E may provide therapeutic benefit for those with autism caused by disturbance of the converging pathways controlling EIF4E activity. PMID- 19556255 TI - Do release-site biases reflect response to the Earth's magnetic field during position determination by homing pigeons? AB - How homing pigeons (Columba livia) return to their loft from distant, unfamiliar sites has long been a mystery. At many release sites, untreated birds consistently vanish from view in a direction different from the home direction, a phenomenon called the release-site bias. These deviations in flight direction have been implicated in the position determination (or map) step of navigation because they may reflect local distortions in information about location that the birds obtain from the geophysical environment at the release site. Here, we performed a post hoc analysis of the relationship between vanishing bearings and local variations in magnetic intensity using previously published datasets for pigeons homing to lofts in Germany. Vanishing bearings of both experienced and naive birds were strongly associated with magnetic intensity variations at release sites, with 90 per cent of bearings lying within +/-29 degrees of the magnetic intensity slope or contour direction. Our results (i) demonstrate that pigeons respond in an orderly manner to the local structure of the magnetic field at release sites, (ii) provide a mechanism for the occurrence of release-site biases and (iii) suggest that pigeons may derive spatial information from the magnetic field at the release site that could be used to estimate their current position relative to their loft. PMID- 19556254 TI - Latitude, elevation and the tempo of molecular evolution in mammals. AB - Faster rates of microevolution have been recorded for plants and marine foraminifera occupying warmer low latitude environments relative to those occurring at higher latitudes. By contrast, because this rate heterogeneity has been attributed to a relationship between thermal habit and mutagenesis via a body temperature linkage, it has been assumed that microevolution in mammals should not also vary systematically with environmental temperature. However, this assumption has not previously been empirically examined. In this study, we tested for a thermally mediated influence on the tempo of microevolution among mammals using a comprehensive global dataset that included 260 mammal species, from 10 orders and 29 families. In contrast to theoretical predictions, we found that substitution rates in the cytochrome b gene have been substantially faster for species living in warmer latitudes and elevations relative to sister species living in cooler habitats. These results could not be attributed to factors otherwise thought to influence rates of microevolution, such as body mass differentials or genetic drift. Instead, the results indicate that the tempo of microevolution among mammals is either responding directly to the thermal environment or indirectly via an ecological mechanism such as the 'Red Queen' effect. PMID- 19556256 TI - Effects of ocean acidification on the early life history of a tropical marine fish. AB - Little is known about how fishes and other non-calcifying marine organisms will respond to the increased levels of dissolved CO(2) and reduced sea water pH that are predicted to occur over the coming century. We reared eggs and larvae of the orange clownfish, Amphiprion percula, in sea water simulating a range of ocean acidification scenarios for the next 50-100 years (current day, 550, 750 and 1030 ppm atmospheric CO(2)). CO(2) acidification had no detectable effect on embryonic duration, egg survival and size at hatching. In contrast, CO(2) acidification tended to increase the growth rate of larvae. By the time of settlement (11 days post-hatching), larvae from some parental pairs were 15 to 18 per cent longer and 47 to 52 per cent heavier in acidified water compared with controls. Larvae from other parents were unaffected by CO(2) acidification. Elevated CO(2) and reduced pH had no effect on the maximum swimming speed of settlement-stage larvae. There was, however, a weak positive relationship between length and swimming speed. Large size is usually considered to be advantageous for larvae and newly settled juveniles. Consequently, these results suggest that levels of ocean acidification likely to be experienced in the near future might not, in isolation, significantly disadvantage the growth and performance of larvae from benthic spawning marine fishes. PMID- 19556257 TI - Mycelial carton galleries of Azteca brevis (Formicidae) as a multi-species network. AB - Apart from growing fungi for nutrition, as seen in the New World Attini, ants cultivate fungi for reinforcement of the walls of their nests or tunnel-shaped runway galleries. These fungi are grown on organic material such as bark, epiphylls or trichomes, and form stable 'carton structures'. In this study, the carton of the runway galleries built by Azteca brevis (Formicidae, Dolichoderinae) on branches of Tetrathylacium macrophyllum (Flacourtiaceae) is investigated. For the first time, molecular tools are used to address the biodiversity and phylogenetic affinities of fungi involved in tropical ant carton architecture, a previously neglected ant-fungus mutualism. The A. brevis carton involves a complex association of several fungi. All the isolated fungi were unequivocally placed within the Chaetothyriales by DNA sequence data. Whereas five types of fungal hyphae were morphologically distinguishable, our DNA data showed that more species are involved, applying a phylogenetic species concept based on DNA phylogenies and hyphal morphology. In contrast to the New World Attini with their many-to-one (different ant species-one fungal cultivar) pattern, and temperate Lasius with a one-to-two (one ant species-two mutualists) or many-to-one (different ant species share the same mutualist) system, the A. brevis-fungi association is a one-to-many multi-species network. Vertical fungus transmission has not yet been found, indicating that the A. brevis-fungi interaction is rather generalized. PMID- 19556258 TI - Enhanced expression of haemoglobin scavenger receptor in accumulated macrophages of culprit lesions in acute coronary syndromes. AB - AIMS: Effective clearance of extracellular haemoglobin (Hb) is thought to limit systemic oxidative heme toxicity, which is presumed to contribute to the pathogenesis of plaque instability. We immunohistochemically examined the relationship between intraplaque haemorrhage, 4-HNE (4-hydroxy-2-nonenal), an index of lipid peroxidation, and the Hb scavenger receptor (CD163), using coronary atherectomy specimens from 74 patients with stable angina pectoris (SAP, n = 39) or unstable angina pectoris (UAP, n = 35). METHODS AND RESULTS: Atherectomy samples were stained with antibodies against glycophorin A (a protein specific to erythrocyte membranes), CD31, 4-HNE, and CD163. Quantitative analysis demonstrated that glycophorin A-positive areas, 4-HNE-positive macrophage score, and CD163-positive macrophage score in UAP patients were significantly higher (glycophorin A, P < 0.0001; 4-HNE-positive macrophage score, P < 0.0001; CD163 positive macrophage score, P < 0.0005) than in SAP patients. The percentage of the glycophorin A-positive area showed a significant positive correlation with the number of CD31-positive microvessels and the 4-HNE-positive macrophage score (microvessels, R = 0.59, P < 0.0001; 4-HNE, R = 0.59, P < 0.0001). Moreover, the CD163-positive macrophage score was positively correlated with glycophorin A positive area and the 4-HNE-positive macrophage score (glycophorin A, R = 0.58, P < 0.0001; 4-HNE, R = 0.53, P < 0.0001). CONCLUSION: These findings suggest a positive association among intraplaque haemorrhage, enhanced expression of Hb scavenger receptor, and lipid peroxidation in human unstable plaques. PMID- 19556259 TI - Recurrent vasovagal syncope: comparison between clomipramine and nitroglycerin as drug challenges during head-up tilt testing. AB - AIMS: To compare the responses between clomipramine, a centrally acting substance, and nitroglycerin, with mainly peripheral action, when each drug is used during tilt test for the induction of vasovagal syncope (VVS). METHODS AND RESULTS: Hundred patients with recurrent episodes of classical VVS underwent two tilt tests in a randomized sequence. One test included 20 min of tilt at 60 degrees with intravenous administration of 5 mg clomipramine (clomipramine tilt), whereas the other test included an initial 30 min period of passive 60 degrees tilt, followed by sublingual spray administration of 400 microg nitroglycerin (nitroglycerin tilt). Fifty asymptomatic subjects served as controls. Following clomipramine tilt, a positive response occurred in 73 patients (73%), a negative response in 23 (23%), and drug intolerance in 4 (4%). With nitroglycerin tilt, these percentages were 52, 48, and 0%, respectively. Significant differences were observed regarding positive responses (clomipramine vs. nitroglycerin: 73/100 vs. 52/100, P < 0.05), as well as negative responses (23/100 vs. 48/100, respectively, P < 0.05). A high concordance rate was observed in positive responses. CONCLUSION: In the evaluation of patients with recurrent classical VVS, clomipramine tilt is associated with an increased positive yield relative to nitroglycerin tilt. This suggests that central mechanisms may be more important than peripheral ones in VVS pathogenesis. PMID- 19556260 TI - Gastrointestinal bleeding in high risk survivors of myocardial infarction: the VALIANT Trial. AB - AIMS: The risk of gastrointestinal (GI) bleeding limits the use of antiplatelet and anticoagulant drugs. Risk factors for GI bleeding in post- myocardial infarction (MI) patients have not been well defined. We sought to identify risk factors for GI bleeding in patients following MI. METHODS AND RESULTS: The VALsartan In Acute myocardial iNfarcTion trial (VALIANT) enrolled 14 703 post-MI patients with left ventricular dysfunction and/or heart failure and followed them for a median of 24.7 months. In the present secondary analysis, times from baseline to first GI bleeding were identified from the VALIANT serious adverse event database. Potential risk factors were explored from medical history, demographics, clinical profile, and medications, both at baseline and during follow-up. We also explored the relationship between the occurrence of GI bleeding and subsequent mortality. During follow-up, 98 (0.7%) patients had a serious GI bleeding event. These patients were older, had more comorbidities, were more likely to be taking additional antiplatelet drugs, and had worse left ventricular systolic and renal function. The Kaplan-Meier estimated rate of GI bleeding at 6 months was 0.37% (95% CI 0.27-0.47). In a multivariable Cox model, dual antiplatelet therapy was the most powerful predictor of GI bleeding, with an adjusted hazard ratio of 3.18 (95% CI 1.91-5.29). Other predictors were non-white race, history of alcohol abuse, increasing age, worse New York Heart Association class, anticoagulant therapy, diabetes, lower estimated glomerular filtration rate, and male sex. Gastrointestinal bleeding was associated with increased risk of death [adjusted hazard ratio 2.54 (95% CI 1.66-3.89)]. CONCLUSION: Following MI, clinical characteristics can identify patients with increased risk of GI bleeding. The use of dual antiplatelet agents appears to be the most profound risk factor. Whether these patients would benefit from GI prophylaxis therapy remains unknown. PMID- 19556261 TI - Delayed neutrophil apoptosis in patients with unstable angina: relation to C reactive protein and recurrence of instability. AB - AIMS: To investigate spontaneous polymorphonuclear neutrophils (PMNs) apoptosis in unstable angina (UA) and its association with recurrence of instability. METHODS AND RESULTS: We compared PMNs apoptotic rate at 4 and 24 h in patients with UA, stable angina (SA), and controls (H) with two different protocols by flow cytometry. We measured apoptotic rate of isolated PMNs (Protocol 1) in 30 UA patients, 13 SA patients, and 34 H; and apoptosis of PMNs in whole blood culture (Protocol 2) in further 10 UA patients, 7 SA patients, and 6 H. Serum high sensitivity C-reactive protein was also measured. Polymorphonuclear neutrophils of UA patients showed a decreased apoptotic rate compared with SA patients and H at 4 h in Protocol 1 (both P < 0.01), and at 24 h in Protocol 2 (P < 0.05 and <0.01, respectively). In overall population, a negative correlation was found between apoptotic rate at 4 h and high-sensitivity C-reactive protein levels (P < 0.01). Six among 40 patients with UA had early recurrence of symptoms and their apoptotic rate was significantly reduced compared with UA patients without recurrence of symptoms (P = 0.024). CONCLUSIONS: Our study demonstrates delayed PMN apoptosis in UA. This alteration might be involved in the persistence of inflammatory activation and affects recurrence of instability. PMID- 19556262 TI - Randomized study on the efficacy of immunosuppressive therapy in patients with virus-negative inflammatory cardiomyopathy: the TIMIC study. AB - AIMS: To evaluate the efficacy of immunosuppression in virus-negative inflammatory cardiomyopathy. METHODS AND RESULTS: This randomized, double-blind, placebo-controlled study included 85 patients with myocarditis and chronic (>6 months) heart failure unresponsive to conventional therapy, with no evidence of myocardial viral genomes. Patients received either prednisone 1 mg kg(-1) day(-1) for 4 weeks followed by 0.33 mg kg(-1) day(-1) for 5 months and azathioprine 2 mg kg(-1) day(-1) for 6 months (43 patients, Group 1) or placebo (42 patients, Group 2) in addition to conventional therapy for heart failure. Primary outcome was the 6 month improvement in left-ventricular function. Group 1 showed a significant improvement of left-ventricular ejection fraction and a significant decrease in left-ventricular dimensions and volumes compared with baseline. None of Group 2 patients showed improvement of ejection fraction, that significantly worsened compared with baseline. No major adverse reaction was registered as a result of immunosuppression. CONCLUSION: These data confirm the efficacy of immunosuppression in virus-negative inflammatory cardiomyopathy. Lack of response in 12% of cases suggests the presence of not screened viruses or mechanisms of damage and inflammation not susceptible to immunosuppression. PMID- 19556263 TI - Ancient and current gene flow between two distantly related Mediterranean oak species, Quercus suber and Q. ilex. AB - BACKGROUND AND AIMS: Quercus suber and Q. ilex are distantly related and their distributions partially overlap. They hybridize occasionally, but the complete replacement of Q. suber chloroplast DNA (cpDNA) by that of Q. ilex was identified in two specific geographical areas. The objective of this study was to determine whether the contrasting situation reflected current or recent geographical interspecies gene flow variation or was the result of ancient introgression. METHODS: cpDNA PCR-RFLPs (restriction fragment length polymorphisms) and variation at ten nuclear microsatellite loci were analysed in populations of each species, in 16 morphologically intermediate individuals and the progeny of several of them. Interspecies nuclear introgression was based on individual admixture rates using a Bayesian approach with no a priori species assignment, and on a maximum-likelihood (ML) method, using allele frequencies in the allopatric populations of each species as controls. Gene flow was compared specifically between populations located within and outside the specific areas. KEY RESULTS: High interspecies nuclear genetic differentiation was observed, with twice the number of alleles in Q. ilex than in Q. suber. According to Bayesian assignment, approx. 1 % of individuals had a high probability of being F(1) hybrids, and bidirectional nuclear introgression affected approx. 4 % of individuals in each species. Hybrid and introgressed individuals were identified predominantly in mixed stands and may have a recent origin. Higher proportions including allospecific genes recovered from past hybridization were obtained using the ML method. Similar rates of hybridization and of nuclear introgression, partially independent of cpDNA interspecies transfer suggestive of gene filtering, were obtained in the populations located within and outside the areas of complete cpDNA replacement. CONCLUSIONS: The results did not provide evidence for geographical variation in interspecies gene flow. In contrast, historical introgression is supported by palynological records and constitutes the more reliable origin of cpDNA replacement in specific regions. PMID- 19556264 TI - Dendrochronological potential of the alpine shrub Rhododendron nivale on the south-eastern Tibetan Plateau. AB - BACKGROUND AND AIMS: Shrubs and dwarf shrubs are wider spread on the Tibetan Plateau than trees and hence offer a unique opportunity to expand the present dendrochronological network into extreme environments beyond the survival limit of trees. Alpine shrublands on the Tibetan Plateau are characterized by rhododendron species. The dendrochronological potential of one alpine rhododendron species and its growth response to the extreme environment on the south-east Tibetan Plateau were investigated. METHODS: Twenty stem discs of the alpine snowy rhododendron (Rhododendron nivale) were collected close to the tongue of the Zuoqiupu Glacier in south-east Tibet, China. The skeleton plot technique was used for inter-comparison between samples to detect the growth pattern of each stem section. The ring-width chronology was developed by fitting a negative exponential function or a straight line of any slope. Bootstrapping correlations were calculated between the standard chronology and monthly climate data. KEY RESULTS: The wood of snowy rhododendron is diffuse-porous with evenly distributed small-diameter vessels. It has well-defined growth rings. Most stem sections can be visually and statistically cross-dated. The resulting 75-year long standard ring-width chronology is highly correlated with a timberline fir chronology about 200 km apart, providing a high degree of confidence in the cross dating. The climate/growth association of alpine snowy rhododendron and of this timberline fir is similar, reflecting an impact of monthly mean minimum temperatures in November of the previous year and in July during the year of ring formation. CONCLUSIONS: The alpine snowy rhododendron offers new research directions to investigate the environmental history of the Tibetan Plateau in those regions where up to now there was no chance of applying dendrochronology. PMID- 19556265 TI - Response to non-uniform salinity in the root zone of the halophyte Atriplex nummularia: growth, photosynthesis, water relations and tissue ion concentrations. AB - BACKGROUND AND AIMS: Soil salinity is often heterogeneous, yet the physiology of halophytes has typically been studied with uniform salinity treatments. An evaluation was made of the growth, net photosynthesis, water use, water relations and tissue ions in the halophytic shrub Atriplex nummularia in response to non uniform NaCl concentrations in a split-root system. METHODS: Atriplex nummularia was grown in a split-root system for 21 d, with either the same or two different NaCl concentrations (ranging from 10 to 670 mm), in aerated nutrient solution bathing each root half. KEY RESULTS: Non-uniform salinity, with high NaCl in one root half (up to 670 mm) and 10 mm in the other half, had no effect on shoot ethanol-insoluble dry mass, net photosynthesis or shoot pre-dawn water potential. In contrast, a modest effect occurred for leaf osmotic potential (up to 30 % more solutes compared with uniform 10 mm NaCl treatment). With non-uniform NaCl concentrations (10/670 mm), 90 % of water was absorbed from the low salinity side, and the reduction in water use from the high salinity side caused whole plant water use to decrease by about 30 %; there was no compensatory water uptake from the low salinity side. Leaf Na(+) and Cl(-) concentrations were 1.9- to 2.3 fold higher in the uniform 670 mm treatment than in the 10/670 mm treatment, whereas leaf K(+) concentrations were 1.2- to 2.0-fold higher in the non-uniform treatment. CONCLUSIONS: Atriplex nummularia with one root half in 10 mm NaCl maintained net photosynthesis, shoot growth and shoot water potential even when the other root half was exposed to 670 mm NaCl, a concentration that inhibits growth by 65 % when uniform in the root zone. Given the likelihood of non-uniform salinity in many field situations, this situation would presumably benefit halophyte growth and physiology in saline environments. PMID- 19556266 TI - Mineral nutrient uptake from prey and glandular phosphatase activity as a dual test of carnivory in semi-desert plants with glandular leaves suspected of carnivory. AB - BACKGROUND AND AIMS: Ibicella lutea and Proboscidea parviflora are two American semi-desert species of glandular sticky plants that are suspected of carnivory as they can catch small insects. The same characteristics might also hold for two semi-desert plants with glandular sticky leaves from Israel, namely Cleome droserifolia and Hyoscyamus desertorum. The presence of proteases on foliar hairs, either secreted by the plant or commensals, detected using a simple test, has long been considered proof of carnivory. However, this test does not prove whether nutrients are really absorbed from insects by the plant. To determine the extent to which these four species are potentially carnivorous, hair secretion of phosphatases and uptake of N, P, K and Mg from fruit flies as model prey were studied in these species and in Roridula gorgonias and Drosophyllum lusitanicum for comparison. All species examined possess morphological and anatomical adaptations (hairs or emergences secreting sticky substances) to catch and kill small insects. METHODS: The presence of phosphatases on foliar hairs was tested using the enzyme-labelled fluorescence method. Dead fruit flies were applied to glandular sticky leaves of experimental plants and, after 10-15 d, mineral nutrient content in their spent carcasses was compared with initial values in intact flies after mineralization. KEY RESULTS: Phosphatase activity was totally absent on Hyoscyamus foliar hairs, a certain level of activity was usually found in Ibicella, Proboscidea and Cleome, and a strong response was found in Drosophyllum. Roridula exhibited only epidermal activity. However, only Roridula and Drosophyllum took up nutrients (N, P, K and Mg) from applied fruit flies. CONCLUSIONS: Digestion of prey and absorption of their nutrients are the major features of carnivory in plants. Accordingly, Roridula and Drosophyllum appeared to be fully carnivorous; by contrast, all other species examined are non carnivorous as they did not meet the above criteria. PMID- 19556268 TI - Bilateral L1 and L2 dorsal root ganglion blocks for discogenic low-back pain. AB - BACKGROUND: It is possible that interruption of nociceptive input from intervertebral discs can be modulated through bilateral L1 and L2 dorsal root ganglia (DRG) blockade. In order to test this hypothesis, we prospectively collected data from patients with low-lumbar pain, accurately diagnosed as discogenic using provocation discography. METHODS: Twelve patients were recruited with a mean (sd) symptom duration of 13.7 (8.2) years. Bilateral DRG blocks of L1 and L2 were performed using methylprednisolone 80 mg, clonidine 75 microg and 0.5% bupivacaine 4 ml in each patient. RESULTS: Analysis of Brief Pain Inventories showed no significant change in pain scores. CONCLUSION: We conclude that blocks of this nociceptive pathway in humans using bilateral DRG blocks has no therapeutic value. PMID- 19556269 TI - Prolonged hoarseness and arytenoid cartilage dislocation after tracheal intubation. AB - BACKGROUND: Hoarseness is a common complication after tracheal intubation and prolonged hoarseness may be very limiting for a patient. This study was designed to examine the duration of hoarseness after tracheal intubation and to identify risk factors that may increase the duration of hoarseness. METHODS: We prospectively studied 3093 adult patients (aged 18-77 yr), over a 3 yr period who required tracheal intubation. Postoperative hoarseness was assessed on the day of operation and on postoperative days 1, 3, and 7 by standardized interview by the resident anaesthetist managing the patient. If postoperative hoarseness was still present on postoperative day 7, the patient was followed up until complete resolution. We evaluated age, gender, weight, Cormack grades, duration of intubation, and the anaesthetic agents used as factors affecting the duration of hoarseness after tracheal intubation. RESULTS: Hoarseness was observed in 49% of patients on the day of surgery and in 29%, 11%, and 0.8% on 1, 3, and 7 postoperative days, respectively. Multiple regression analysis showed that patient age and duration of intubation, but not gender, weight, Cormack grades, or the agents used, were significant predictors of increased duration of hoarseness after tracheal intubation. We found three patients with arytenoid cartilage dislocation (0.097%) in our study population. CONCLUSIONS: The age of the patient and duration of intubation were significant factors in the duration of hoarseness after tracheal intubation. In addition, the incidence of arytenoid cartilage dislocation was 0.097%. PMID- 19556267 TI - Nitric oxide and frataxin: two players contributing to maintain cellular iron homeostasis. AB - BACKGROUND: Nitric oxide (NO) is a signalling and physiologically active molecule in animals, plants and bacteria. The specificity of the molecular mechanism(s) involved in transducing the NO signal within and between cells and tissues is still poorly understood. NO has been shown to be an emerging and potent signal molecule in plant growth, development and stress physiology. The NO donor S nitrosoglutathion (GSNO) was shown to be a biologically active compound in plants and a candidate for NO storage and/or mobilization between plant tissues and cells. NO has been implicated as a central component in maintaining iron bioavailavility in plants. SCOPE AND CONCLUSIONS: Iron is an essential nutrient for almost all organisms. This review presents an overview of the functions of NO in iron metabolism in animals and discusses how NO production constitutes a key response in plant iron sensing and availability. In plants, NO drives downstream responses to both iron deficiency and iron overload. NO-mediated improvement of iron nutrition in plants growing under iron-deficient conditions represents a powerful tool to cope with soils displaying low iron availability. An interconversion between different redox forms based on the iron and NO status of the plant cells might be the core of a metabolic process driving plant iron homeostasis. Frataxin, a recently identified protein in plants, plays an important role in mitochondria biogenesis and in maintaining mitochondrial iron homeostasis. Evidence regarding the interaction between frataxin, NO and iron from analysis of frataxin knock-down Arabidopsis thaliana mutants is reviewed and discussed. PMID- 19556270 TI - Meperidine-induced serotonin syndrome in a susceptible patient. AB - We present a patient with a history of clomipramine-induced serotonin syndrome 5 yr prior who developed serotonin syndrome after a single dose of meperidine. This report heightens appreciation of population at risk and also recognition of potential toxicity in meperidine. PMID- 19556271 TI - Anaesthesia for deep brain stimulation and in patients with implanted neurostimulator devices. AB - Deep brain stimulation has become an increasingly common treatment for Parkinson's disease and other movement disorders. Consequently, it is important to understand the concepts of appropriate patient selection, the implantation process, and the various drugs and techniques that can be used to facilitate this treatment. Currently, none of the anaesthetic techniques for neurostimulator implantation has proven to be superior to others, although awake or sedation techniques are popular as they facilitate intraoperative neurological testing. However, even with meticulous anaesthetic care, perioperative complications such as hypertension and seizures do occasionally occur and close monitoring is required. Anaesthesia in patients with an implanted neurostimulator requires special considerations because of possible interference between neurostimulators and other devices. We have reviewed the current knowledge of anaesthetic techniques and perioperative complications of neurostimulator insertion. Anaesthetic considerations in patients with an implanted neurostimulator are also discussed. PMID- 19556272 TI - Training and the European Working Time Directive: a 7 year review of paediatric anaesthetic trainee caseload data. AB - BACKGROUND: The implementation of the European Working Time Directive (WTD) has reduced the hours worked by trainees in the UK to a maximum of 56 h per week. With a further and final reduction to 48 h per week scheduled for August 2009, there is concern amongst doctors about the impact on training and on patient care. Paediatric anaesthesia is one of the specialist areas of anaesthesia for which the Royal College of Anaesthetists (RCoA) recommends a minimum caseload during the period of advanced training. METHODS: We conducted a retrospective analysis of theatre logbook data from 62 Specialist Registrars (SpRs) who had completed a 12 month period of advanced training in paediatric anaesthesia in our institution between 2000 and 2007. RESULTS: After the implementation of the WTD 56 h week in 2004, the mean total number of cases performed by SpRs per year decreased from 441 to 336, a 24% reduction. We found a statistically significant reduction across all age groups with the largest reduction in the under 1 month of age group. The post-WTD group did not meet the RCoA recommended total minimum caseload or the minimum number of cases of <1 yr of age. CONCLUSIONS: Since the implementation of the WTD, there has been a significant reduction in the number of cases performed by SpRs in paediatric anaesthesia and they are no longer achieving the RCoA recommended minimum numbers for advanced training. PMID- 19556273 TI - Russia's healthcare system: in need of modernisation. PMID- 19556274 TI - Cost of US health reforms is estimated at $1.6 trillion for first 10 years. PMID- 19556275 TI - Making information about clinical trials publicly available. PMID- 19556278 TI - New analysis pinpoints ethnic differences in cancer incidence in England. PMID- 19556279 TI - Unique ion signature mass spectrometry, a deterministic method to assign peptide identity. AB - The growing use of selected reaction monitoring (SRM) mass spectrometry in proteomic analyses led us to investigate how to identify peptides by SRM using only a minimal number of fragment ions. By using a computational model of the SRM work flow we computed the potential interferences from other peptides in a given proteome. From these results, we selected the deterministic SRM addresses that contained sufficient information to confer peptide and protein identity that we termed unique ion signatures (UIS). We computationally showed that UIS comprised of only two transitions are diagnostic for >99% of Escherichia coli proteins and >96% of human proteins that possess a sequence-unique peptide. We demonstrated an example of experimental use of UIS using a modified SRM methodology to profile the E. coli tricarboxylic acid cycle from a single injection of cell lysate. In addition, we showed the potential of UIS to form the first functionally orthogonal approach to validate peptide assignments obtained from conventional analyses of MS/MS spectra. The UIS methodology is a novel deterministic peptide identification method for MS/MS spectra based on information content. These robust theoretical assays will have widespread use when integrated with previously collected MS/MS data and conventional proteomics technologies. PMID- 19556280 TI - Reverse rate dependency is an intrinsic property of canine cardiac preparations. AB - AIMS: Class III antiarrhythmic agents exhibit reverse rate-dependent lengthening of the action potential duration (APD). In spite of the several theories developed so far to explain this reverse rate dependency (RRD), its mechanism has not yet been clarified. The aim of the present work was to further elucidate the mechanisms responsible for reverse rate-dependent drug effects. METHODS AND RESULTS: Action potentials were recorded from multicellular canine ventricular preparations and isolated cardiomyocytes, at cycle lengths (CLs) varying from 0.3 to 5 s, using conventional sharp microelectrodes. APD was either modified by applying inward and outward current pulses, or by superfusion of agents known to lengthen and shorten APD. Net membrane current (I(m)) was calculated from action potential waveforms. The hypothesis that RRD may be implicit in the relationship between I(m) and APD was tested by numerical modelling. Both drug-induced lengthening (by veratrine, BAY-K 8644, dofetilide, and BaCl(2)) and shortening (by lidocaine and nicorandil) of action potentials displayed RRD, i.e. changes in APD were greater at longer than at shorter CL. A similar dependency of effect on CL was found when repolarization was modified by injection of inward or outward current pulses. I(m) measured at various points during repolarization was inversely proportional to APD and to CL. Model simulations showed that RRD is expected as a consequence of the non-linearity of the relationship between I(m) and APD. CONCLUSION: RRD of APD modulation is shared, although with differences in magnitude, by interventions of very different nature. RRD can be interpreted as a consequence of the relationship between I(m) and APD and, as such, is expected in all species having positive APD-CL relationship. This implies that the development of agents prolonging APD with direct rate dependency, or even completely devoid of RRD, may be difficult to achieve. PMID- 19556286 TI - Emerging roles of chemokines in prostate cancer. AB - Prostate cancer (PCa) represents the second leading cause of death among all cancer types in men in Europe and North America. Among the factors suspected to control PCa, incidence and progression, chemokines, and their receptors are now intensively studied. Chemokines are produced by tumor cells and also by the stromal microenvironment, both in the primary tumor site and in distant metastatic locations. The wide and differential distribution of chemokines and their receptors account for the pleiotropic actions of chemokines in PCa, including the modulation of growth, angiogenesis, invasion, metastasis, and hormone escape. This review will focus on the roles and the mechanisms of action and regulation of chemokines in the different steps of PCa development and will discuss the novel strategies that are currently envisioned to target chemokines in PCa. PMID- 19556287 TI - Expression of aryl hydrocarbon receptor (AHR) and AHR-interacting protein in pituitary adenomas: pathological and clinical implications. AB - Germline mutations of the aryl hydrocarbon receptor (AHR)-interacting protein (AIP) gene confer a predisposition to pituitary adenomas (PA), usually in the setting of familial isolated PA. To provide further insights into the possible role of AIP in pituitary tumour pathogenesis, the expression of AIP and AHR was determined by real-time RT-PCR and/or immunohistochemistry (IHC) in a large series of PA (n=103), including 17 with AIP mutations (AIP(mut)). Variable levels of AIP and AHR transcripts were detected in all PA, with a low AHR expression (P<0.0001 versus AIP). Cytoplasmic AIP and AHR were detected by IHC in 84.0 and 38.6% of PA respectively, and significantly correlated with each other (P=0.006). Nuclear AHR was detected in a minority of PA (19.7%). The highest AIP expression was observed in somatotrophinomas and non-secreting (NS) PA, and multivariate analysis in somatotrophinomas showed a significantly lower AIP immunostaining in invasive versus non-invasive cases (P=0.019). AIP expression was commonly low in other secreting PA. AIP immunostaining was abolished in a minority of AIP(mut) PA, with a frequent loss of cytoplasmic AHR and no evidence of nuclear AHR. In contrast, AIP overexpression in a subset of NS PA could be accompanied by nuclear AHR immunopositivity. We conclude that down-regulation of AIP and AHR may be involved in the aggressiveness of somatotrophinomas. Overall, IHC is a poorly sensitive tool for the screening of AIP mutations. Data obtained on AHR expression suggest that AHR signalling may be differentially affected according to PA phenotype. PMID- 19556288 TI - Immortalized human skin fibroblast feeder cells support growth and maintenance of both human embryonic and induced pluripotent stem cells. AB - BACKGROUND: Feeder cells are frequently used for the early-stage of derivation and culture of human embryonic stem cell (hESC) lines. METHODS: We established a conditionally immortalized human foreskin fibroblast line that secreted basic fibroblast growth factor (bFGF). These cells were used as feeder cells for hESC culture and induced pluripotent stem (iPS) cell derivation and expansion. This conditional immortalization was performed using lentiviral vector (LV) mediated transduction of Bmi-1 and human telomerase reverse transcriptase genes and the resulting cell line was further modified by LV-mediated transduction of a secreted form of bFGF gene product. Three different laboratories have tested whether this feeder cell line could support the maintenance of four different hESC lines. RESULTS: Immortalized fibroblasts secreting stable amounts of bFGF supported the growth of all hESC lines, which remained pluripotent and had a normal karyotype for at least 10 passages. Even at high passage (p56), these modified cells, when used as feeders, could support iPS derivation and propagation. Derived iPS cells expressed pluripotency markers, had hESC morphology and produced tissue components of the three germ layers when differentiated in vitro. CONCLUSION: These modified fibroblasts are useful as a genetically-defined feeder cell line for reproducible and cost-effective culture of both hESC and iPS cells. PMID- 19556289 TI - Proteomic analysis of the human receptive versus non-receptive endometrium using differential in-gel electrophoresis and MALDI-MS unveils stathmin 1 and annexin A2 as differentially regulated. AB - BACKGROUND: The transcriptome of the endometrium throughout the menstrual cycle has been described in recent years. However, the proteomic of the window of implantation remains unknown. The aim of this study was to compare the proteome of the human endometrium in the pre-receptive phase versus the receptive phase by identifying and quantifying the proteins differentially expressed using differential in-gel electrophoresis (DIGE) and mass spectometry (MS). METHODS: Endometrial biopsies were collected at days 2 (pre-receptive) and 7 (receptive) after the urinary luteal hormone surge in the same menstrual cycle from eight fertile women (corresponding to days 16 and 21 of the menstrual cycle). Proteins were extracted and labeled with CyDye DIGE fluorofores and separated using two dimensional gel electrophoresis. RESULTS: Image analysis using the DeCyder software followed by protein identification by matrix-assisted laser desorption/ionization-MS and database searching revealed 32 differentially expressed proteins, although only annexin A2 and stathmin 1 were consistently regulated in the two experiments performed. Validation and localization of annexin A2 and stathmin 1 were performed by western blot and immunohistochemistry. Annexin A2 and stathmin 1 were investigated using an endometrial refractoriness model. The results highlight the key potential of these proteins as possible targets for human endometrial receptivity and interception. CONCLUSION: This study shows that the human endometrium has a differential proteomic repertoire during the window of implantation. Consequently, we identified annexin A2 and stathmin 1 as differentially expressed molecules in the receptive endometrium. PMID- 19556290 TI - Chlamydia trachomatis YtgA is an iron-binding periplasmic protein induced by iron restriction. AB - Chlamydia trachomatis is a Gram-negative obligate intracellular bacterium that is the causative agent of common sexually transmitted diseases and the leading cause of preventable blindness worldwide. It has been observed that YtgA (CT067) is very immunogenic in patients with chlamydial genital infections. Homology analyses suggested that YtgA is a soluble periplasmic protein and a component of an ATP-binding cassette (ABC) transport system for metals such as iron. Since little is known about iron transport in C. trachomatis, biochemical assays were used to determine the potential role of YtgA in iron acquisition. (59)Fe binding and competition studies revealed that YtgA preferentially binds iron over nickel, zinc or manganese. Western blot and densitometry techniques showed that YtgA concentrations specifically increased 3-5-fold in C. trachomatis, when cultured under iron-starvation conditions rather than under general stress conditions, such as exposure to penicillin. Finally, immuno-transmission electron microscopy provided evidence that YtgA is more concentrated in C. trachomatis during iron restriction, supporting a possible role for YtgA as a component of an ABC transporter. PMID- 19556291 TI - Small heat-shock protein HspL is induced by VirB protein(s) and promotes VirB/D4 mediated DNA transfer in Agrobacterium tumefaciens. AB - Agrobacterium tumefaciens is a Gram-negative plant-pathogenic bacterium that causes crown gall disease by transferring and integrating its transferred DNA (T DNA) into the host genome. We characterized the chromosomally encoded alpha crystallin-type small heat-shock protein (alpha-Hsp) HspL, which was induced by the virulence (vir) gene inducer acetosyringone (AS). The transcription of hspL but not three other alpha-Hsp genes (hspC, hspAT1, hspAT2) was upregulated by AS. Further expression analysis in various vir mutants suggested that AS-induced hspL transcription is not directly activated by the VirG response regulator but rather depends on the expression of VirG-activated virB genes encoding components of the type IV secretion system (T4SS). Among the 11 virB genes encoded by the virB operon, HspL protein levels were reduced in strains with deletions of virB6, virB8 or virB11. VirB protein accumulation but not virB transcription levels were reduced in an hspL deletion mutant early after AS induction, implying that HspL may affect the stability of individual VirB proteins or of the T4S complex directly or indirectly. Tumorigenesis efficiency and the VirB/D4-mediated conjugal transfer of an IncQ plasmid RSF1010 derivative between A. tumefaciens strains were reduced in the absence of HspL. In conclusion, increased HspL abundance is triggered in response to certain VirB protein(s) and plays a role in optimal VirB protein accumulation, VirB/D4-mediated DNA transfer and tumorigenesis. PMID- 19556292 TI - The cellular level of O-antigen polymerase Wzy determines chain length regulation by WzzB and WzzpHS-2 in Shigella flexneri 2a. AB - The lipopolysaccharide O antigen of Shigella flexneri 2a has two preferred chain lengths, a short (S-OAg) composed of an average of 17 repeated units and a very long (VL-OAg) of about 90 repeated units. These chain length distributions are controlled by the chromosomally encoded WzzB and the plasmid-encoded Wzz(pHS-2) proteins, respectively. In this study, genes wzzB, wzz(pHS-2) and wzy (encoding the O-antigen polymerase) were cloned under the control of arabinose- and rhamnose-inducible promoters to investigate the effect of varying their relative expression levels on O antigen polysaccharide chain length distribution. Controlled expression of the chain length regulators wzzB and wzz(pHS-2) revealed a dose-dependent production of each modal length. Increase in one mode resulted in a parallel decrease in the other, indicating that chain length regulators compete to control the degree of O antigen polymerization. Also, when expression of the wzy gene is low, S-OAg but not VL-OAg is produced. Production of VL-OAg requires high induction levels of wzy. Thus, the level of expression of wzy is critical in determining O antigen modal distribution. Western blot analyses of membrane proteins showed comparable high levels of the WzzB and Wzz(pHS-2) proteins, but very low levels of Wzy. In vivo cross-linking experiments and immunoprecipitation of membrane proteins did not detect any direct interaction between Wzy and WzzB, suggesting the possibility that these two proteins may not interact physically but rather by other means such as via translocated O antigen precursors. PMID- 19556293 TI - Proteolytic degradation of human salivary MUC5B by dental biofilms. AB - The degradation of complex substrates, like salivary mucins, requires an arsenal of glycosidases and proteases to sequentially degrade the oligosaccharides and polypeptide backbone. The mucin MUC5B is a complex oligomeric glycoprotein, heterogeneous in molecular mass (14-40 x 10(6) Da), with a diverse repertoire of oligosaccharides, differing in composition and charge. The aim of this study was to investigate whether proteolytic degradation of the mucin polypeptide backbone could be identified and if cooperation of dental biofilm bacteria was required. Cooperative bacteria-mediated proteolysis of MUC5B was determined by comparing individual species and mixed consortia of strains isolated from supragingival plaque, and freshly harvested supragingival plaque. Proteolytic activity was analysed using fluorescent labelled substrate and by visualizing mucin degradation by SDS-PAGE. Dental plaque degraded the polypeptide backbone of the salivary MUC5B mucin. The mucin was also degraded by a specific consortium of isolated species from supragingival plaque, although individual species and other consortia did not. Certain bacteria in supragingival dental plaque therefore cooperate as a consortium to proteolyse human salivary MUC5B and hydrolyse glycosides. PMID- 19556294 TI - Characterization of a multimeric, eukaryotic prolyl aminopeptidase: an inducible and highly specific intracellular peptidase from the non-pathogenic fungus Talaromyces emersonii. AB - Fungi are capable of degrading proteins in their environment by secreting peptidases. However, the link between extracellular digestion and intracellular proteolysis has scarcely been investigated. Mycelial lysates of the filamentous fungus Talaromyces emersonii were screened for intracellular peptidase production. Five distinct proteolytic activities with specificity for the p nitroanilide (pNA) peptides Suc-AAPF-pNA, Suc-AAA-pNA, K-pNA, F-pNA and P-pNA were identified. The native enzyme responsible for the removal of N-terminal proline residues was purified to homogeneity by ammonium sulfate fractionation followed by five successive chromatographic steps. The enzyme, termed Talaromyces emersonii prolyl aminopeptidase (TePAP), displayed a 50-fold specificity for cleaving N-terminal Pro-X (k(cat)/K(m)=2.1 x 10(6) M(-1) s(-1)) compared with Ala X or Val-X bonds. This intracellular aminopeptidase was optimally active at pH 7.4 and 50 degrees C. Peptide sequencing facilitated the design of degenerate oligonucleotides from homologous sequences encoding putative fungal proline aminopeptidases, enabling subsequent cloning of the gene. TePAP was shown to be relatively uninhibited by classical serine peptidase inhibitors and to be sensitive to selected cysteine- and histidine-modifying reagents, yet gene sequence analysis identified the protein as a serine peptidase with an alpha/beta hydrolase fold. Northern analysis indicated that Tepap mRNA levels were regulated by the composition of the growth medium. Highest Tepap transcript levels were observed when the fungus was grown in medium containing glucose and the protein hydrolysate casitone. Interestingly, both the induction profile and substrate preference of this enzyme suggest potential co-operativity between extracellular and intracellular proteolysis in this organism. Gel filtration chromatography suggested that the enzyme exists as a 270 kDa homo-hexamer, whereas most bacterial prolyl aminopeptidases (PAPs) are monomers. Phylogenetic analysis of known PAPs revealed two diverse subfamilies that are distinguishable on the basis of primary and secondary structure and appear to correlate with the subunit composition of the native enzymes. Sequence comparisons revealed that PAPs with key conserved topological features are widespread in bacterial and fungal kingdoms, and this study identified many putative PAP candidates within sequenced genomes. This work represents, to our knowledge, the first detailed biochemical and molecular analysis of an inducible PAP from a eukaryote and the first intracellular peptidase isolated from the thermophilic fungus T. emersonii. PMID- 19556296 TI - Nitric oxide mediates the fungal-elicitor-enhanced biosynthesis of antioxidant polyphenols in submerged cultures of Inonotus obliquus. AB - A fungal elicitor prepared from the cell debris of the plant-pathogenic ascomycete Alternaria alternata induces multiple responses by Inonotus obliquus cells, including an increase in generation of nitric oxide (NO), activity of phenylalanine ammonia lyase (PAL) and accumulation of total mycelial phenolic compounds (TMP), but does not trigger production of oxylipins or jasmonic acid (JA). The role of NO in TMP production was investigated via the effects of the NO specific scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3 oxide (cPITO) and the nitric oxide synthase (NOS) inhibitor aminoguanidine (AG). TMP profiles were assayed using (1)H NMR spectroscopy combining multivariate pattern recognition strategies. Pretreatment of I. obliquus mycelia with cPITO or AG suppressed not only elicitor-enhanced NO generation and PAL activity, but also the elicitor-induced increase in TMP production. This TMP reduction by either a NO scavenger or a NOS inhibitor was reversed by exogenous addition of either a NO donor, sodium nitroprusside, or JA separately. NMR-based metabonomic analysis of TMP profiles showed that the induced TMP were hispidin analogues including inoscavins, phelligridins, davallialactone and methyldavallialactone, which possess high antioxidant activities. Thus, NO mediates an elicitor-induced increase in production of antioxidant polyphenols in I. obliquus via a signalling pathway independent of oxylipins or JA, a mechanism which differs from those in some higher plants. PMID- 19556295 TI - Mycobacteriophages BPs, Angel and Halo: comparative genomics reveals a novel class of ultra-small mobile genetic elements. AB - Mycobacteriophages BPs, Angel and Halo are closely related viruses isolated from Mycobacterium smegmatis, and possess the smallest known mycobacteriophage genomes, 41,901 bp, 42,289 bp and 41,441 bp, respectively. Comparative genome analysis reveals a novel class of ultra-small mobile genetic elements; BPs and Halo each contain an insertion of the proposed mobile elements MPME1 and MPME2, respectively, at different locations, while Angel contains neither. The close similarity of the genomes provides a comparison of the pre- and post-integration sequences, revealing an unusual 6 bp insertion at one end of the element and no target duplication. Nine additional copies of these mobile elements are identified in a variety of different contexts in other mycobacteriophage genomes. In addition, BPs, Angel and Halo have an unusual lysogeny module in which the repressor and integrase genes are closely linked. The attP site is located within the repressor-coding region, such that prophage formation results in expression of a C-terminally truncated, but active, form of the repressor. PMID- 19556297 TI - Simultaneous occurrence of hepatitis C virus-associated glomerulonephritis and AL amyloidosis. AB - We report a 65-year-old male with hepatitis C virus (HCV)-associated glomerulonephritis (GN) and AL amyloidosis. This patient had a 14-year history of HCV infection with positive serum HCV RNA at presentation. Diagnosis of HCV related GN was established using immunohistochemistry in which the HCV-NS3 antigen was mainly detected as granular deposition in glomerular mesangium. AL amyloidosis was determined by electron microscopy, positive Congo red staining and identification of lambda-chain in the kidney specimen and monoclonal IgG lambda in serum and urine. Under immunoelectron microscopy, the HCV-NS3 antigen was found in electron-dense deposits, while lambda-chains appeared in the amyloid like filaments. PMID- 19556298 TI - Saturated fatty acids induce insulin resistance in human podocytes: implications for diabetic nephropathy. AB - BACKGROUND: Cellular insulin resistance is the hallmark of type 2 diabetes and predominantly affects adipose and muscle cells. The saturated free fatty acid palmitate is elevated in insulin-resistant states and may directly contribute to cellular insulin resistance. A spectrum of renal disease is associated with increased markers of insulin resistance, although direct causal mechanisms are not known. In the kidney, glomerular podocytes are novel insulin-sensitive cells that have the ability to rapidly transport glucose. In this study, we tested the hypothesis that palmitate would induce insulin resistance in podocytes. METHODS: Conditionally immortalized human podocytes were cultured for up to 24 h with 375 750 muM palmitate. Functional effects on glucose uptake and ceramide production were measured. Gene expression was investigated using a focused gene array, and protein signalling and trafficking were studied with Western blotting and immunofluorescence. RESULTS: We found that palmitate blocked insulin-stimulated glucose uptake in human podocytes. This was associated with increased ceramide production, and use of the ceramide inhibitors myriocin and fumonisin B1 partially recovered the insulin sensitivity. At the level of transcription, palmitate downregulated genes associated with several pathways involved in insulin signalling. At the protein level, phosphorylation of the insulin receptor, IRS1 and PKB was reduced and there was impaired translocation of GLUT4 to the cell surface. CONCLUSION: This is the first study to demonstrate a direct effect of saturated fatty acids on podocyte function. These findings may represent a novel link between systemic insulin resistance and the development of nephropathy. PMID- 19556299 TI - Safety and efficacy of percutaneous insertion of peritoneal dialysis catheters under sedation and local anaesthetic. AB - BACKGROUND: Success of peritoneal dialysis (PD) is partially dependent on the ease of insertion of the catheter. We have been inserting PD catheters percutaneously in a majority of our patients under local anaesthetic and sedation by physicians, and detail here the outcomes for 283 catheters inserted in this manner, and 150 patients with surgical catheter insertion by laparotomy or laparoscopy. METHODS: Data were collected prospectively on all patients having PD catheters inserted between 1999 and 2008, including success of insertion, complications and infections. RESULTS: A total of 283 catheters were inserted percutaneously using a Seldinger technique under sedation and local anaesthesia, and 150 surgically under general anaesthetic. Eighty-six percent of the percutaneous catheters and 66% surgical catheters were first catheters. No major complications occurred. In 7% of the percutaneous patients and 5% surgical patients, the procedure failed or was abandoned. Poor initial drainage occurred in 21% insertions but resolved in most cases and resolved dialysate leak in 6%. Wound infections or peritonitis occurred in 9% and 4% of percutaneous insertions. Only 13% of patients could not use their catheter at 1 month after percutaneous insertion, and 83% of the patients remained on PD using the original catheter at 6 months. CONCLUSIONS: Percutaneous PD catheter insertion was associated with a very low complication rate and high primary success rate, and was highly efficient in use of resources and avoided the need for general anaesthesia. PMID- 19556300 TI - IgA nephropathy associated with ankylosing spondylitis is not controlled by infliximab therapy. AB - The association between seronegative spondyloarthro- pathies and IgA nephropathy is well documented, mainly in cases of ankylosing spondylitis (AS). However, although these diseases have been associated, the physiopathological links between each other appear unclear. Anti-TNFalpha agents have transformed the outcome of axial forms of AS resistant to conventional anti-inflammatory therapies. Infliximab, a monoclonal anti-TNFalpha antibody, has greatly improved the evolution of AS although several adverse events have been described. On the other hand, infliximab has been demonstrated to reduce renal symptoms associated with chronic inflammatory rheumatological diseases, such as amyloid A (AA) amyloidosis, but few data are available on its efficacy in controlling IgA nephropathy associated with AS [1,2]. We report here a case of IgA nephropathy associated with AS that became symptomatic, whereas infliximab therapy efficiently controlled the rheumatological disease. This suggests that even though infliximab therapy effectively controls rheumatological manifestations, it may not be able to prevent IgA nephropathy associated with AS. Thus, this case report illustrates the complexity of the physiopathology of both diseases. PMID- 19556301 TI - Renal recovery after injury: the role of Pax-2. PMID- 19556302 TI - Antimicrobial cathelicidin polypeptide CAP11 suppresses the production and release of septic mediators in D-galactosamine-sensitized endotoxin shock mice. AB - Endotoxin shock is a severe systemic inflammatory response that is caused by the augmented production and release of septic mediators. Among them, inflammatory cytokines such as tumor necrosis factor-alpha, IL-1beta and IL-6 play a pivotal role. In addition, anandamide, an endogenous cannabinoid and high-mobility group box-1 (HMGB1), a non-histone chromosomal protein has recently been recognized as members of septic mediators. We previously reported that cationic antibacterial polypeptide of 11-kDa (CAP11), an antimicrobial cathelicidin peptide (originally isolated from guinea pig neutrophils), potently neutralizes the biological activity of LPS and protects mice from lethal endotoxin shock. In this study, to clarify the protective mechanism of CAP11 against endotoxin shock, we evaluated the effects of CAP11 on the production and release of septic mediators in vitro and in vivo using a murine macrophage cell line RAW264.7 and a D-galactosamine sensitized murine endotoxin shock model. LPS stimulation induced the production of inflammatory cytokines and anandamide and release of HMGB1 from RAW264.7 cells. Importantly, CAP11 suppressed the LPS-induced production and release of these mediators by RAW264.7 cells. Moreover, LPS administration enhanced the serum levels of HMGB1, anandamide and inflammatory cytokines in the endotoxin shock model. Of note, CAP11 suppressed the LPS-induced increase of these mediators in sera, and LPS binding to CD14-positive cells (peritoneal macrophages), accompanied with the increase of survival rates. Together these observations suggest that the protective action of CAP11 on endotoxin shock may be explained by its suppressive effect on the production and release of septic mediators by CD14-positive cells possibly via the inhibition of LPS binding to the targets. PMID- 19556303 TI - Anti-moesin antibodies derived from patients with aplastic anemia stimulate monocytic cells to secrete TNF-alpha through an ERK1/2-dependent pathway. AB - Antibodies specific to moesin, which are frequently detectable in the serum of patients with aplastic anemia (AA), can induce tumor necrosis factor-alpha (TNF alpha) secretion from monocytes and a human monocytic leukemia cell line THP-1. We investigated the mechanisms responsible for TNF-alpha secretion from monocytic cells induced by the auto-antibodies that are purified from the sera of AA patients. TNF-alpha induction by anti-moesin antibodies depended on the amount of cell surface moesin expressed by THP-1 cells. F(ab')(2) fragments prepared from the anti-moesin antibodies were able to stimulate THP-1 cells to secrete TNF alpha and this stimulatory effect was enhanced by cross-linking of moesins with anti-human IgG F(ab')(2) fragment antibodies. Anti-moesin antibodies as well as their F(ab')(2) fragments induced the phosphorylation of ERK1/2 in monocytic cells and this effect was suppressed by the addition of an ERK1/2 inhibitor. Moreover, anti-moesin antibody treatment induced the phosphorylation of moesin proteins in the monocytes and THP-1 cells within 30 min. These results indicate that anti-moesin antibodies induce TNF-alpha secretion from monocytes through the activation of the ERK1/2 pathway provoked by direct binding to moesin on the cells. PMID- 19556304 TI - Gene expression profiling of experimental asthma reveals a possible role of paraoxonase-1 in the disease. AB - In this study, we aimed to identify novel genes involved in experimental and human asthma, importance of which has not yet been recognized. In an ovalbumin induced murine model of asthma, we applied microarray gene expression analysis at different time points after allergen challenges. Advanced statistical methods were used to relate gene expression changes to cellular processes and to integrate our results into multiple levels of information available in public databases. At 4 h after the first allergen challenge, gene expression pattern reflected mainly an acute, but non-atopic, inflammatory response and strong chemotactic activity. At 24 h after the third allergen challenge, gene set enrichment analysis revealed significant over-representation of gene sets corresponding to T(h)2-type inflammation models. Among the top down-regulated transcripts, an anti-oxidant enzyme, paraoxonase-1 (PON1), was identified. In human asthmatic patients, we found that serum PON1 activity was reduced at exacerbation, but increased parallel with improving asthma symptoms. PON1 gene polymorphisms did not influence the susceptibility to the disease. Our observations suggest that an altered PON1 activity might be involved in the pathogenesis of asthma, and serum PON1 level might be used for following up the effect of therapy. PMID- 19556305 TI - Nucleotide-hydrolyzing antibodies from the sera of autoimmune-prone MRL-lpr/lpr mice. AB - Abzymes (Abzs) with different enzymic activities have been detected in the sera of patients with various autoimmune (AI) diseases and in AI mice. In this work, electrophoretically homogeneous IgGs were isolated from the sera of MRL-lpr/lpr mice spontaneously developing lupus-like AI pathology. It was shown for the first time that polyclonal IgGs (pIgGs) and their isolated heavy and light chains hydrolyze different nucleoside-5'-triphosphate (NTPs), nucleoside-5'-diphosphate (NDPs), adenosine monophosphate and deoxiadenosine-5'-monophosphate (dAMP), whereas antibodies from the sera of control healthy mice were catalytically inactive. Monoclonal mouse IgGs also effectively hydrolyze nucleotides. The data demonstrate that nucleotide-hydrolyzing activity is an intrinsic property of isolated mouse pIgG and monoclonal IgG. It was shown that various markers of AI pathologies (proteinuria and antibody titers to native and denatured DNA) demonstrating spontaneous development of AI reactions increased in animals with aging and correlated with an increase in Abz relative activity in hydrolysis of nucleotides. The highest increase in AI reaction markers and in Abz enzymic activity was found in mice immunized with a DNA-protein complex. PMID- 19556306 TI - Mutational analysis of Cys(88) of Toll-like receptor 4 highlights the critical role of MD-2 in cell surface receptor expression. AB - The role of MD-2 in cell surface expression of Toll-like receptor (TLR) 4 has been controversial. The purposes of this study were to characterize the N-glycan of TLR4 and to investigate the roles of MD-2 in N-linked glycosylation and cell surface expression of TLR4. Lectin blot and cell surface biotinylation revealed that TLR4 exhibited the 110 kDa protein with high mannose type N-glycans and the 130 kDa protein with complex type N-glycans and that only the 130 kDa TLR4 with complex type N-glycans was expressed on the cell surface. The cells transfected with a mutant TLR4(C88A) alone expressed only the 110 kDa TLR4 with a high mannose type N-glycan, which did not appear on the cell surface. However, TLR4(C88A) acquired complex type N-glycans and was expressed on the cell surface when MD-2 was co-transfected. The amount of the 130 kDa TLR4(C88A) with complex type N-glycans expressed on the cell surface depended on that of MD-2 transfected. alpha-Mannosidase II inhibitor blocked the processing N-glycans to complex type, but TLR4 with high mannose type appeared on the cell surface, suggesting that TLR4 is destined to locate on the cell surface before processing N-glycans from a high mannose type to a complex type. From these results, we conclude that MD-2 is critical for cell surface expression of TLR4(C88A). This study provides evidence that MD-2 possesses potential ability to play an essential role in cell surface expression of TLR4. PMID- 19556307 TI - Apex2 is required for efficient somatic hypermutation but not for class switch recombination of immunoglobulin genes. AB - The DNA cleavage step in both the class switch recombination (CSR) and somatic hypermutation (SHM) of Ig genes is initiated by activation-induced cytidine deaminase (AID). However, the detailed mechanisms of the DNA strand cleavage in SHM and CSR are still largely unknown. Recently, the apurinic/apyrimidinic endonucleases, Apex1 and Apex2, were reported to be involved in the DNA cleavage step of CSR. Here, we examined the role of Apex2 in SHM using Apex2-deficient mice and found that the Apex2 deficiency caused a drastic reduction in the frequency of SHM and the number of mutations per mutated clone without affecting the pattern of base substitution. These results suggest that Apex2 may play a critical role in SHM through its 3'-5' exonuclease activity. Unexpectedly, the efficiency of CSR was not reduced in Apex2-deficient B cells. In addition, Apex1 knockdown in CH12F3-2 B lymphoma cells did not affect the CSR frequency, suggesting that neither Apex1 nor Apex2 plays a major role in CSR. PMID- 19556308 TI - The CD70-CD27 interaction during the stimulation with dendritic cells promotes naive CD4(+) T cells to develop into T cells producing a broad array of immunostimulatory cytokines in humans. AB - CD70 expressed on dendritic cells (DCs) has been shown to play a critical role in inducing effective CD8(+) T cell responses and a T(h)1 response in mice. However, it has not been extensively examined whether human primary DCs express CD70 and whether the CD70-CD27 interaction promotes naive CD4(+) T cells to acquire the ability to produce effector cytokines during the DC-T cell interaction in humans. Here, we show that human myeloid dendritic cells (mDCs) and plasmacytoid dendritic cells stimulated with CD40 ligand together with pro-inflammatory cytokines or Toll-like receptor ligands express CD70. Thymic stromal lymphopoietin plus prostaglandin E(2) also induced CD70 on mDCs. Naive CD4(+) T cells stimulated with DCs but not with anti-CD3/CD28 microbeads expressed CD70. Stimulation with CD70 together with anti-CD3/CD28 microbeads imparted the ability to produce T(h)1 (IFN-gamma), T(h)2 (IL-4, IL-5, IL-13) cytokines, IL-2 and tumor necrosis factor-alpha to naive CD4(+) T cells. The production of IFN-gamma was associated with the induction of T-bet. Naive CD4(+) T cells stimulated with mDCs acquired an enhanced ability to produce a broad array of immunostimulatory cytokines in a CD70-dependent manner. These data suggest that human CD70 expressed on mDCs and activated T cells transmits a 'basal level' signal, rather than a 'polarizing' signal, to naive CD4(+) T cells, in that CD70 promotes the development of CD4(+) T cells that produce a variety of effector cytokines including both T(h)1 and T(h)2 types, thus contributing to the enhancement of a broad spectrum of immune responses. PMID- 19556309 TI - To report or not report? Attitudes held by Norwegian nursing home staff on reporting inadequate care carried out by colleagues. AB - AIMS: The aims of this study are, first, to describe attitudes held by nursing home staff on reporting acts of inadequate care committed by their colleagues, and second, to investigate whether nursing staff have different attitudes depending on age, education, and length of experience of working in the healthcare services. METHODS: A questionnaire survey was conducted among nursing staff in 16 nursing homes in the central part of Norway. The response rate was 79% (n=616). RESULTS: A positive attitude towards reporting acts of inadequate care committed by their colleagues was held by the participants in this study. Compared with younger staff, the older staff seemed to be more reluctant to report colleagues, to feel less brave, to be more afraid of what would happen to them if they reported, and to agree that it is best to deal with such matters internally. Regarding education, it seemed that a higher educational level was related with a more positive attitude towards a willingness to report and less fear of negative sanctions. CONCLUSIONS: Staff who observe acts of inadequate care committed by colleagues agree that it is their intention to report such incidents. Institutions need to develop and implement mechanisms for understanding and evaluating acts of inadequate care, and staff must be encouraged to speak out on behalf of residents rather than be punished for doing so. PMID- 19556310 TI - Work life and mental wellbeing of single and non-single working mothers in Scandinavia. AB - BACKGROUND: This study examined levels and predictors of mental wellbeing in Scandinavian working single and non-single mothers, with a special focus on financial stress, job characteristics and work-family conflict. METHODS: The European Social Survey Round 2 (2005) provided questionnaire data from 73 single and 432 non-single working mothers in Denmark, Sweden and Norway. Respondents answered questions about the outcome variables life satisfaction, happiness, and positive affect, and predictor variables financial stress, job characteristics, work-family conflict, and social support. Hierarchical multiple regression was used to assess the relationships between predictor variables and mental wellbeing outcomes. RESULTS: Single working mothers scored significantly lower on life satisfaction and happiness, but not on positive affect, than did non-single mothers. Financial stress was higher in the single mother group. There were no significant differences in levels of enriching or stressful job characteristics, or in levels of social support. While financial stress and work-family conflict were important predictors in both groups, the relationship between financial stress and wellbeing was far stronger in the single mother group. Confidant support was a significant predictor only in the single mother group, and social participation only in the non-single mothers group. CONCLUSIONS: This study suggests that the Scandinavian welfare democracies have not yet been successful in relieving the financial pressure experienced by single working mothers. Development of efficient financial support systems should be prioritized. Ways to reduce work-family conflict in both single and non-single mothers in Scandinavia should also be given increased attention. PMID- 19556311 TI - Sensitivity of conventional memory tests in multiple sclerosis: comparing the Rao Brief Repeatable Neuropsychological Battery and the Minimal Assessment of Cognitive Function in MS. AB - BACKGROUND: Cognitive impairment is common in multiple sclerosis (MS) affecting roughly 45-60% of patients. Because memory deficits have significant impact on employment, caregiver burden, and social functioning, neuropsychological (NP) assessment is often recommended. Two widely used and validated NP batteries for MS are the Rao Brief Repeatable Neuropsychological Battery (BRNB) and the Minimal Assessment of Cognitive Function in MS (MACFIMS). Although similar, these batteries differ in the specific auditory/verbal and visual/spatial memory tests employed. The relative sensitivity of these memory tests is unknown. METHODS: The BRNB and MACFIMS have considerable overlap but different memory tests: the former includes the Selective Reminding Test (SRT) and the 10/36 Spatial Recall Test (10/36) and the latter the California Verbal Learning Test, Second Edition (CVLT2) and Brief Visuospatial Memory Test, Revised (BVMTR). In 65 patients with MS and 46 demographically matched controls, we compared the sensitivity of these tests, and secondarily their respective batteries. RESULTS: The BRNB and MACFIMS were comparable in their overall sensitivity to disease status. Although the BVMTR showed greater discriminative validity than the 10/36, the CVLT2 and SRT were comparable in sensitivity. The SDMT was the most sensitive NP test across both batteries. CONCLUSIONS: We conclude that the BRNB and MACFIMS have comparable sensitivity among patients with MS. The sensitivity of the auditory/verbal memory tests from these batteries is similar, but the BVMTR appears to be more sensitive than the 10/36. Clinical implications are discussed. PMID- 19556312 TI - Home administration of intravenous methylprednisolone for multiple sclerosis relapses: the experience of French multiple sclerosis networks. AB - BACKGROUND: One single center study has provided support for a home-based approach to the therapeutic management of multiple sclerosis (MS) relapse. OBJECTIVE: To report a multicenter series of patients with MS who were treated at home for a relapse with a 3-day course of intravenous methylprednisolone. METHODS: The home administration of intravenous methylprednisolone was coordinated by four MS networks in France; patients with MS with a relapse were referred by their neurologists, and treatment was administered by a local nurse. We analyzed the safety and efficiency of this approach and estimated the related cost savings. Patients completed a patient satisfaction questionnaire. RESULTS: Eight hundred and seven patients received intravenous methylprednisolone at home. The mean disease duration was 10.3 +/- 7.9 years. Treatment was often prescribed by community-based neurologists. The delay between prescription and treatment was 2.8 +/- 0.5 days if treatment was initiated at home and 1.9 +/- 3.0 days if treatment was initiated in hospital (the subsequent two injections were always administered at home). Home treatment was well tolerated; three serious side effects requiring hospital transfer were observed (anxiety, thoracic oppression, and arrhythmia), which were fully reversible. Overall, 93.8% of patients were satisfied with the treatment approach, and 98% wished to receive future treatment courses at home. The overall cost savings of home-based treatment versus hospital based treatment were evaluated at EUR1,091,482. CONCLUSION: Safety data, patient satisfaction, and economic considerations support home-based treatment of MS relapses with intravenous methylprednisolone, provided strict patient selection criteria are observed and the process is coordinated and closely monitored by an MS network. PMID- 19556313 TI - Cognitive dysfunction in primary progressive multiple sclerosis: a neuropsychological and MRI study. AB - Although cognitive dysfunction is known to occur in multiple sclerosis (MS), only few studies have reported cognitive performance in patients with primary progressive MS (PPMS). To find out the pattern of cognitive performance in PPMS, 28 PPMS patients underwent an extensive battery of neuropsychological tests. The results were compared to those of healthy controls (n = 20) and patients with secondary progressive MS (SPMS, n = 28). Furthermore, the results of neuropsychological tests in PPMS were correlated to magnetic resonance imaging findings. Our study showed that the PPMS patients have deficits in several cognitive domains when compared to age-matched and education-matched controls, but the cognitive impairment in the PPMS and SPMS patients appeared to be similar. Cognitive deficits in PPMS patients correlated with diffuse brain lesion, T1- and T2-lesion load, but no correlations were found with atrophy. PMID- 19556314 TI - The development and validation of the Unidimensional Fatigue Impact Scale (U FIS). AB - BACKGROUND: The multidimensional assessment of fatigue is complicated by the interrelation of its multiple causes and effects. OBJECTIVE: The purpose of the research was to develop a unidimensional assessment of fatigue (U-FIS). METHODS: Data collected with the Fatigue Impact Scale (FIS) were subjected to Rasch analysis to identify potential problems with the scale. Additional items for the U-FIS were generated from interviews with UK MS patients. The U-FIS was tested for face and content validity in patient interviews and included in a validation survey to determine dimensionality (Rasch model), reliability and validity. RESULTS: The original FIS was not unidimensional when subscale items were combined. The modification of the FIS and addition of a number of items allowed the development of a 22-item unidimensional scale (U-FIS) that was reliable (Cronbach Alpha = 0.96; test-retest = 0.86,) and valid given correlations with the Nottingham Health Profile and ability to distinguish between MS severity groups. There was no significant difference in U-FIS scores according to MS type. CONCLUSION: It is valid to conceptualize the functional impact of fatigue as unidimensional. The U-FIS is a reliable and valid questionnaire that will allow the measurement of this construct in clinical studies. PMID- 19556315 TI - The development of patient-reported outcome indices for multiple sclerosis (PRIMUS). AB - BACKGROUND: Complex diseases such as multiple sclerosis (MS) present dilemmas over the choice of patient-reported outcome measures as no single scale can inform on all types of MS impact from the patient's perspective. OBJECTIVE: To develop an outcome tool, the Patient-Reported Indices for Multiple Sclerosis (PRIMUS), to assess MS symptoms, activities, and quality of life. METHODS: PRIMUS content was derived from qualitative interviews with UK MS patients and checked by clinical experts. Semi-structured cognitive debriefing interviews assessed scale face and content validity. PRIMUS scaling properties, reliability, and construct validity were assessed by a test-retest postal survey. RESULTS: Cognitive debriefing interviews (n = 15) demonstrated scale clarity, relevance, and comprehensiveness. The postal survey was completed by 135 patients with MS. After removal of misfitting items and those exhibiting differential item functioning, all scales fitted the Rasch model, confirming unidimensionality. For all scales, test-retest reliability exceeded 0.80. Scale scores were related to perceived MS severity, general health, and symptoms of depression. Moderate correlations were observed between PRIMUS and Nottingham Health Profile scores. CONCLUSIONS: Clinicians and researchers can have confidence in scores obtained by respondents on the PRIMUS. The PRIMUS will aid the assessment of the impact of MS from the patient's perspective. PMID- 19556316 TI - Quantitative assessment of brain iron by R(2)* relaxometry in patients with clinically isolated syndrome and relapsing-remitting multiple sclerosis. AB - BACKGROUND: Increased iron deposition has been implicated in the pathophysiology of multiple sclerosis (MS), based on visual analysis of signal reduction on T(2) weighted images. R(2)* relaxometry allows to assess brain iron accumulation quantitatively. OBJECTIVE: To investigate regional brain iron deposition in patients with a clinically isolated syndrome (CIS) or relapsing-remitting MS (RRMS) and its associations with demographical, clinical, and conventional magnetic resonance imaging (MRI) parameters. METHODS: We studied 69 patients (CIS, n = 32; RRMS, n = 37) with 3T MRI and analyzed regional R(2)* relaxation rates and their correlations with age, disease duration, disability, T(2) lesion load, and normalized brain volumes. RESULTS: Basal ganglia R(2)* relaxation rates increased in parallel with age (r = 0.3-0.6; P < 0.01) and were significantly higher in RRMS than in CIS (P < 0.05). Using multivariate linear regression analysis, the rate of putaminal iron deposition was independently predicted by the patients' age, disease duration, and gray matter atrophy. CONCLUSIONS: Quantitative assessment by R(2)* relaxometry suggests increased iron deposition in the basal ganglia of MS patients, which is associated with disease duration and brain atrophy. This technique together with long-term follow-up thus appears suited to clarify whether regional iron accumulation contributes to MS morbidity or merely reflects an epiphenomenon. PMID- 19556317 TI - Glypican 5 is an interferon-beta response gene: a replication study. AB - BACKGROUND: Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system. Interferon-beta is the most usual therapy in relapsing remiting MS. However, approximately 50% of the treated patients do not respond adequately. Very recently, a genome-wide association study on interferon-beta pharmacogenetics has described polymorphisms at several genes that are associated with response to this treatment. Our aim is to replicate the results obtained at the two loci most strongly implicated in the response to interferon-beta treatment, HAPLN1 and GPC5. PATIENTS AND METHODS: We performed a case-control study, analyzing 199 patients with MS treated with interferon-beta for at least 2 years and at least two documented relapses over the 2 years, previous to treatment onset. Responders had neither relapses nor increase in expanded disability status scale (EDSS) over the 2-year follow-up period, whereas nonresponders had at least two relapses or an increase in EDSS of at least 1 point. We studied three single-nucleotide polymorphisms (SNPs) in the GPC5 locus and three SNPs in the HAPLN1 locus by TaqMan technology. Allelic frequencies between responders and nonresponders were compared by a chi-square test. RESULTS: An association was found between GPC5 polymorphisms and the response to interferon-beta therapy in patients with MS, in agreement with earlier data (responder vs nonresponder patients: rs10492503, P = 0.0005). The other locus studied (HAPLN1) did not show association with treatment response to interferon beta (all SNPs P > 0.05). CONCLUSIONS: We confirm the association of polymorphisms within GPC5 with response to interferon-beta therapy in patients with MS. PMID- 19556318 TI - Phase II randomized study of trabectedin given as two different every 3 weeks dose schedules (1.5 mg/m2 24 h or 1.3 mg/m2 3 h) to patients with relapsed, platinum-sensitive, advanced ovarian cancer. AB - BACKGROUND: This randomized, open-label, phase II clinical trial evaluated the optimal regimen of trabectedin administered every 3 weeks in patients with platinum-sensitive, relapsed, advanced ovarian cancer (AOC). PATIENTS AND METHODS: Patients previously treated with less than two or two previous chemotherapy lines were randomized to receive trabectedin 1.5 mg/m(2) 24 h (arm A, n = 54) or 1.3 mg/m(2) 3 h (arm B, n = 53). Objective response rate (ORR) per RECIST was the primary efficacy end point. Toxic effects were graded according to the National Cancer Institute-Common Toxicity Criteria v. 2.0. RESULTS: ORR was 38.9% [95% confidence interval (CI) 25.9% to 53.1%; arm A] and 35.8% (95% CI 23.1% to 50.2%; arm B) (intention-to-treat primary analysis). Median time to progression was 6.2 months (95% CI 5.3-8.6 months; arm A) and 6.8 months (95% CI 4.6-7.4 months; arm B). Frequent severe adverse events were nausea/vomiting (24%, arm A; 15%, arm B) and fatigue (15%, arm A; 10%, arm B). Common severe laboratory abnormalities were transient, noncumulative neutropenia (55%, arm A; 37%, arm B) and transaminase increases (alanine aminotransferase, 55%, arm A; 59%, arm B). CONCLUSIONS: Both every-3-weeks trabectedin regimes, 1.5 mg/m(2) 24 h and 1.3 mg/m(2) 3 h, were active and reasonably well tolerated in AOC platinum-sensitive patients. Trabectedin every-3-weeks has promising activity and deserves to be further evaluated in relapsed AOC. PMID- 19556319 TI - Disseminated tumor cells and the risk of locoregional recurrence in nonmetastatic breast cancer. AB - BACKGROUND: In early breast cancer patients, bone marrow (BM)-disseminated tumor cells (DTCs) were associated with distant metastasis and locoregional recurrence. Our aim was to determine whether BM DTC detection could be related to specific locoregional dissemination of cancer cells, according to radiotherapy volumes. PATIENTS AND METHODS: The relationship between locoregional recurrence-free survival (LRFS) and DTC detection was evaluated according to the various locoregional volumes irradiated after surgery. RESULTS: BM DTCs were detected in 94 of 621 stage I-III breast cancer patients (15%) and were not associated with axillary node status. Eighteen patients (2.9%) experienced locoregional recurrence (median follow-up 56 months), of whom eight (44%) were initially BM DTC positive. BM DTC detection was the only prognostic factor for LRFS [P = 0.0005, odds ratio = 5.2 (2.0-13.1), multivariate analysis]. In BM DTC-positive patients, a longer LRFS was observed in those who were given adjuvant hormone therapy (P = 0.03) and radiotherapy to supraclavicular nodes (SCNs)/internal mammary nodes (IMNs) (P = 0.055) (multivariate analysis; interaction test: P = 0.028). CONCLUSIONS: The presence of DTC in BM may be associated with a different pattern of locoregional cancer cell dissemination and influences LRFS. The possible reseeding of the primary cancer area by DTC could be prevented by systemic hormone therapy but also by SCN/IMN irradiation. PMID- 19556320 TI - Circulating tumor cells in metastatic inflammatory breast cancer. AB - BACKGROUND: Inflammatory breast cancer (IBC) is the most aggressive form of breast cancer. Circulating tumor cells (CTCs) are an independent prognostic factor in metastatic breast cancer. The aim of this study was to assess the prognostic value of baseline CTCs in metastatic IBC patients. PATIENTS AND METHODS: This retrospective study included 42 metastatic IBC and 107 metastatic non-IBC patients treated with first- or second-line chemotherapy from January 2004 to December 2007 at MD Anderson Cancer Center. CTCs were detected and enumerated before patients started chemotherapy using the CellSearch system. RESULTS: Ten (23.8%) IBC patients versus 48 (44.9%) non-IBC patients had baseline CTCs > or =5 per 7.5 ml of peripheral blood. IBC patients had a lower mean +/- SEM CTCs than non-IBC patients (7.6 +/- 2.9 versus 34.2 +/- 9.1; P = 0.02). The estimated median overall survival was 26.5 versus 18.3 months (P = 0.68) in IBC patients and 37.4 versus 18.3 months (P = 0.016) in non-IBC patients with CTCs <5 and CTCs > or =5, respectively. CONCLUSIONS: Metastatic IBC patients had a lower prevalence and fewer CTCs in comparison to metastatic non-IBC patients. Survival of metastatic IBC patients with <5 CTCs was not significantly better than that of patients with > or =5 CTCs. Further research is warranted with prospective assessment of CTCs in IBC patients and their biological characterization. PMID- 19556321 TI - Docetaxel and pegylated liposomal doxorubicin combination as first-line therapy for metastatic breast cancer patients: results of the phase II GINECO trial CAPYTTOLE. AB - BACKGROUND: This phase II study evaluated the clinical benefit of pegylated liposomal doxorubicin (PLD) and docetaxel (Taxotere) as first-line therapy for metastatic breast cancer (MBC). PATIENTS AND METHODS: MBC patients were enrolled to receive six cycles of PLD 35 mg/m2 (day 1) and docetaxel 40 mg/m2 (days 1 and 15), every 28 days (group A). Because of unacceptable toxic effects, doses were modified to PLD 30 mg/m2 (day 1) and docetaxel 75 mg/m2 (day 2), every 3 weeks (group B). The primary end point was clinical benefit. RESULTS: Sixty-seven patients were included (group A, 53; group B, 14). In both groups, the median number of cycles delivered was 4 and the overall dose intensity was 82% for docetaxel and 71% for PLD. In group A, main toxic effects were hematologic, palmar-plantar erythrodysesthesia (PPE), and stomatitis. In group B, higher rates of grade 3-4 PPE, febrile neutropenia, and hematologic toxic effects were reported. The rate of clinical benefit was 47%. Among patients with a measurable disease, 49% achieved a partial response, 27% had a stable disease, and 13% progressed, according to RECIST criteria. CONCLUSION: The combination of PLD and docetaxel delivered at planned doses in this study yields unacceptable toxicity and should not be used routinely in patients with MBC. PMID- 19556322 TI - High prevalence of retinoblastoma protein loss in triple-negative breast cancers and its association with a good prognosis in patients treated with adjuvant chemotherapy. AB - BACKGROUND: Triple-negative breast cancer (TNBC) is an aggressive disease, nevertheless exhibiting a high response rate to chemotherapy. Since the retinoblastoma protein (pRb) loss confers a high sensitivity to chemotherapy regimens, we evaluated the prevalence of pRb loss in TNBCs and its relevance on the clinical outcome of patients treated with adjuvant chemotherapy. PATIENTS AND METHODS: pRb status was prospectively evaluated by immunocytochemistry in 518 consecutive patients with complete receptor information. The predictive value of pRb status in TNBCs was determined according to the adjuvant therapeutic treatments. RESULTS: Fifty-three tumors were identified as TNBCs. The prevalence of pRb loss was significantly higher in TNBCs than in the other cancer subtypes. All patients with TNBCs lacking pRb and treated with systemic chemotherapy (cyclophosphamide, methotrexate and 5-fluorouracil) were disease free at a medium follow-up time of 109 months, whereas the clinical outcome of those expressing pRb was significantly poorer (P = 0.008). Analysis of disease-free survival including the established anatomo-clinical prognostic parameters indicated pRb loss as the only significant predictive factor. CONCLUSIONS: pRb loss is much more frequent in TNBCs than in the other breast cancer subtypes. Patients with TNBCs lacking pRb had a very favorable clinical outcome if treated with conventional adjuvant chemotherapy. PMID- 19556323 TI - Quality assurance for radical hysterectomy for cervical cancer: the view of the European Organization for Research and Treatment of Cancer--Gynecological Cancer Group (EORTC-GCG). AB - Radical hysterectomy combined with a pelvic lymphadenectomy or chemoradiation are traditionally the mainstays of treatment of International Federation of Gynecology and Obstetrics stages Ia2-IIa cervical cancer. The quality of radical surgery for cervical cancer influences local tumor control and survival. Hence, it is important to optimize and ensure the quality of surgical care for cervical cancer patients. In this paper, we discuss factors that are related to outcome after radical hysterectomy and propose a set of quality indicators that can be used to audit and improve the quality of surgical care for cervical cancer patients. PMID- 19556324 TI - Predicting the risk of functional decline in older patients admitted to the hospital: a comparison of three screening instruments. PMID- 19556325 TI - Older men with higher self-rated socioeconomic status have shorter telomeres. AB - BACKGROUND: previous studies examining the relationship between socioeconomic status and telomere length showed conflicting results, one study finding shorter telomere length in subjects with lower socioeconomic status and one showing no relationship. DESIGN: cross-sectional study. SETTING: community-living elderly Chinese in Hong Kong. OBJECTIVE: this study examines the relationship between self-rated social economic status and telomere length in Hong Kong Chinese men and women aged 65 years and over living in the community. SUBJECTS AND METHOD: information was collected from 958 men and 978 women regarding possible confounding factors such as the presence of chronic diseases, smoking, physical activity level, dietary intake and body mass index. Telomere length was measured by quantitative PCR. RESULT: in men only, after adjustment for age and other confounding factors, a higher ranking in community standing was associated with shorter telomere length. CONCLUSION: men with higher self-rated socioeconomic status have shorter telomeres, possibly mediated through psychosocial rather than lifestyle factors or the presence of chronic disease. There may be cultural ethnic and age-related differences in social determinants of health. PMID- 19556326 TI - Spontaneous femoral shaft fracture after long-term alendronate. AB - Alendronate is used as first-line treatment for osteoporosis in postmenopausal women worldwide. The evidence supporting its benefit in reducing the risk of fragility fracture among this high-risk group is well established. Due to its mechanism of action, there were concerns about potential reduction in bone turnover, repair of microdamage and hence reduced bone toughness and increased risk of fractures with long-term use. A number of cases have been reported from other countries describing a pattern of atraumatic mid-femoral fractures in women on long-term alendronate therapy. We present a case of an atraumatic femoral shaft fracture in an elderly woman on long-term alendronate therapy admitted to a UK Hospital. PMID- 19556327 TI - Cohort profile: the Chinese national free antiretroviral treatment cohort. PMID- 19556328 TI - Elevated serum creatine kinase predicts first-ever myocardial infarction: a 12 year population-based cohort study in Japan, the Suita study. AB - BACKGROUND: In myocardial infarction (MI), it is well known that serum creatine kinase (s-CK) increases after onset, but it is unclear whether s-CK elevates before MI onset. The present analysis examined whether elevated s-CK levels predicted first-ever MI or stroke. METHODS: This study was a population-based cohort study in a Japanese urban area. Study subjects were comprised of 5026 initially healthy Japanese (2370 men and 2656 women, mean age: 54.5 years) without a history of MI or stroke. They were followed-up for 11.8 years on average, and 103 MIs (definite: 45; probable: 58) and 168 strokes (definite: 126; probable: 42) were observed. There was no subject who developed MI just at baseline (the follow-up period among those with definite MI was, at earliest, 0.20 years). RESULTS: The adjusted hazard ratio for definite MI was 4.18 (95% confidence interval 1.66-10.53) with s-CK levels of >or=200 IU/l, compared with the reference category (s-CK levels of or =.81, indicating high concurrent validity. The predictive validity of the 4 measures was satisfactory (Spearman rho, > or =.51). The responsiveness of the 4 measures at 14 to 180 days after stroke was moderate (.52 < or = effect size < or = .79). The 4 measures had good interrater reliability (intraclass correlation coefficient [ICC], > or =.92) and test-retest reliability (ICC, > or =.97). Only the minimal detectable changes of the UE-FM (8% of the highest possible score) and the ARAT (6%) were satisfactory. LIMITATIONS: The sample size was too small to conduct data analysis according to type or severity of stroke. In addition, the timed component of the Wolf Motor Function Test was not used in this study. CONCLUSIONS: All 4 measures showed sufficient validity, responsiveness, and reliability in participants with stroke. The UE-FM for assessing impairment and the ARAT for assessing disability had satisfactory minimal detectable changes, supporting their utility in clinical settings. PMID- 19556334 TI - Violence and women's mental health: the pain unequalled: a two-part special issue. PMID- 19556335 TI - Human axillary odor: are there side-related perceptual differences? AB - Most studies on perception of human social odors in axillary sweat do not distinguish between samples from the right and left axillae. However, each axilla might not produce identical odor samples due, for instance, to the increased use of one arm as a result of lateralization. The aim of the present study was to test whether odor samples from the right and left axillae provided by right- and left-handed men were perceived differently by female raters. Participants were 38 males and 49 females, aged 19-35 years. Fresh odor samples (cotton pads worn underarm for 24 h) were evaluated for attractiveness, intensity, and masculinity, with left and right samples being presented as independent stimuli. A side related difference emerged in left-handers only (no difference in right-handers): The odor from the axilla corresponding to the dominant side (left) was rated more masculine and more intense than the other side (right). This effect was limited to the ratings of a restricted group of females, that is, those who did not take hormone-based contraception and were estimated to be in the fertile phase of their menstrual cycle. In conclusion, future studies using axillary odor samples can consider left and right samples as perceptually equivalent stimuli when the participant samples are representative of the general population, which comprises relatively low proportions of left-handed men and spontaneously ovulating fertile women. The results also provide new evidence of the variation of female sensitivity to biologically relevant stimuli across the menstrual cycle. PMID- 19556336 TI - Discourse analysis: what is it and why is it relevant to family practice? AB - This paper aims to illustrate what discourse analysis is and how it can contribute to our understanding of family practice. Firstly, we describe what 'discourse analysis' is, mapping the discourse analysis terrain by discussing four studies relevant to primary care to illustrate different methodological approaches and key concepts. We then address the practicalities of how to actually do discourse analysis, providing readers with a worked example using one particular approach. Thirdly, we touch on some common debates about discursive research. We conclude by advocating that researchers and practitioners take up the challenge of understanding, utilizing and extending the field of discourse studies within family practice. PMID- 19556338 TI - Clinical evidences of laparoscopic versus open surgery for colorectal cancer. AB - Laparoscopic surgery has widely spread in the treatment of colorectal cancer. In Japan, a nation-wide survey has shown that a rate of advanced colorectal cancer has increased gradually and reached 65% of the total cases for colorectal cancer in 2007. For colon cancer, many randomized controlled trials regarding short-term outcome demonstrate that laparoscopic surgery is feasible, safe and has many benefits including reduction in a peri-operative mortality. In terms of long-term outcome, four randomized controlled trials insist that there are no differences in both laparoscopic and open surgeries. However, there are still more important issues including long-term oncological outcome for advanced colon cancer, cost effectiveness and the impact on quality of life of patients. Meanwhile, for rectal cancer, a controversy persists with regard to the appropriateness of laparoscopic surgery because of concerns over the safety of the procedure and a necessity of lateral lymph node dissection for lower rectal cancer. At present, laparoscopic surgery is acceptable for Stage I colon cancer, whereas there are controversies for Stage II/III colon cancer and each staged rectal cancer because of inadequate clinical evidences. Whether laparoscopic surgery further spreads to be applied for colorectal cancer or not, it would be confirmed by Japanese large scale phase III trial (JCOG0404) estimating oncological outcome for Stage II/III colon cancer and a Phase II trial estimating the feasibility for Stage 0/I rectal cancer in near future. PMID- 19556337 TI - Allergic reactions to oxaliplatin in a single institute in Japan. AB - Allergic reactions to oxaliplatin can be severe and are an important cause of discontinuation of treatment. A retrospective review was performed for 105 patients who received FOLFOX regimens between May 2005 and June 2007. Twenty-five cases (23.8%) of allergic reactions were identified, including 9 late onset reactions (8.6%) and 16 immediate reactions (15.2%). Severe allergy (Grades 3 and 4) occurred in seven patients (6.7%). Re-introduction of FOLFOX was attempted for seven immediate onset patients with a severity grade of 1 or 2, and three of these patients (42.9%) showed relapse of allergy. In approximately 10% of the patients, FOLFOX had to be discontinued due to allergy before the disease became refractory to the regimen. Our experience indicates that allergy to oxaliplatin may be a significant concern and that methods are required for suppression of this allergy. PMID- 19556339 TI - Clinical impact of the presence of the worst nucleolar grade in renal cell carcinoma specimens. AB - OBJECTIVE: Renal cell carcinoma (RCC) with a high-nucleolar-grade component is considered to be an aggressive type of tumor. In the present study, we evaluated the impact of the presence of the worst-nucleolar-grade component and also tried to determine predictors for recurrence and prognosis in patients with the worst grade component. METHODS: We evaluated 314 patients with RCC. A three-graded system was used for nucleolar grading, the patients were classified into four groups according to the presence of the worst nucleolar grade (Grade 3) and the occupancy of each grade, and clinicopathological factors and clinical outcomes were compared. In patients of Grade 3 components (Groups 1 and 2), factors influencing on prognosis and recurrence were evaluated by multivariate analysis. RESULTS: There was no significant difference in clinicopathological factors between Group 1 (with Grade 3-dominant tumors) and Group 2 (with tumors in which Grade 1 or 2 was dominant and there were Grade 3 components). Neither did cause specific survival or recurrence-free survival differ significantly between those two groups. In multivariate analysis, only distant metastasis was an independent predictor for prognosis in all patients with Grade 3 components. Moreover, an elevated C-reactive protein (CRP) level (>or=1 mg/dl) was the only independent predictor of recurrence in N0M0 patients. CONCLUSIONS: Regardless of dominancy, the presence of the worst grade component has a significant clinical impact in RCC patients. N0M0 patients whose RCC has worst-grade components but whose CRP levels are <1 are expected to have longer recurrence-free intervals and to survive longer than those whose CRP levels are higher. PMID- 19556340 TI - Novel regulators of Fgf23 expression and mineralization in Hyp bone. AB - We used gene array analysis of cortical bone to identify Phex-dependent gene transcripts associated with abnormal Fgf23 production and mineralization in Hyp mice. We found evidence that elevation of Fgf23 expression in osteocytes is associated with increments in Fgf1, Fgf7, and Egr2 and decrements in Sost, an inhibitor in the Wnt-signaling pathway, were observed in Hyp bone. beta-Catenin levels were increased in Hyp cortical bone, and TOPflash luciferase reporter assay showed increased transcriptional activity in Hyp-derived osteoblasts, consistent with Wnt activation. Moreover, activation of Fgf and Wnt-signaling stimulated Fgf23 promoter activity in osteoblasts. We also observed reductions in Bmp1, a metalloproteinase that metabolizes the extracellular matrix protein Dmp1. Alterations were also found in enzymes regulating the posttranslational processing and stability of Fgf23, including decrements in the glycosyltransferase Galnt3 and the proprotein convertase Pcsk5. In addition, we found that the Pcsk5 and the glycosyltransferase Galnt3 were decreased in Hyp bone, suggesting that reduced posttranslational processing of FGF23 may also contribute to increased Fgf23 levels in Hyp mice. With regard to mineralization, we identified additional candidates to explain the intrinsic mineralization defect in Hyp osteoblasts, including increases in the mineralization inhibitors Mgp and Thbs4, as well as increases in local pH-altering factors, carbonic anhydrase 12 (Car12) and 3 (Car3) and the sodium-dependent citrate transporter (Slc13a5). These studies demonstrate the complexity of gene expression alterations in bone that accompanies inactivating Phex mutations and identify novel pathways that may coordinate Fgf23 expression and mineralization of extracellular matrix in Hyp bone. PMID- 19556341 TI - Rapid estradiol/ERalpha signaling enhances aromatase enzymatic activity in breast cancer cells. AB - In situ estrogen production by aromatase conversion from androgens plays an important role in breast tumor promotion. Here, we show that 17beta-estradiol (E2) can rapidly enhance aromatase enzymatic activity through an increase of aromatase protein phosphorylation in breast cancer cell lines. In vivo labeling experiments and site-directed mutagenesis studies demonstrated that phosphorylation of the 361-tyrosine residue is crucial in the up-regulation of aromatase activity under E2 exposure. Our results demonstrated a direct involvement of nonreceptor tyrosine-kinase c-Src in E2-stimulated aromatase activity because inhibition of its signaling abrogated the up-regulatory effects induced by E2 on aromatase activity as well as phosphorylation of aromatase protein. In addition, from our data it emerges that aromatase is a target of cross talk between growth factor receptors and estrogen receptor alpha signaling. These findings show, for the first time, that tyrosine phosphorylation processes play a key role in the rapid changes induced by E2 in aromatase enzymatic activity, revealing the existence of a short nongenomic autocrine loop between E2 and aromatase in breast cancer cells. PMID- 19556342 TI - ASCOM controls farnesoid X receptor transactivation through its associated histone H3 lysine 4 methyltransferase activity. AB - Activating signal cointegrator-2 (ASC-2), a coactivator of multiple nuclear receptors and transcription factors, belongs to a steady-state complex named ASCOM (for ASC-2 complex), which contains histone H3 lysine 4 (H3K4) methyltransferase MLL3 or its paralog MLL4. ASC-2 binds to many nuclear receptors in a ligand-dependent manner through its two LxxLL motifs. Here we show that the first LxxLL motif of ASC-2 shows relatively weak but specific interaction with the nuclear receptor farnesoid X receptor (FXR) and that ASCOM plays crucial roles in FXR transactivation. Our results reveal that ASC-2, MLL3, and MLL4 are recruited to FXR target genes in a ligand-dependent manner. We further show that the recruitment of MLL3 requires ASC-2 and that FXR ligand induces not only expression of FXR-target genes but also their H3K4 trimethylation in a manner dependent on the presence of ASC-2, MLL3, and MLL4. In addition, MLL3 and MLL4 function redundantly with FXR transactivation. Correspondingly, expression of FXR target genes is partially impaired in mice expressing an enzymatically inactivated mutant form of MLL3, and these mice show disrupted bile acid homeostasis. Overall, these results suggest that ASCOM-MLL3 and ASCOM-MLL4 play redundant but essential roles in FXR transactivation via their H3K4 trimethylation activity. PMID- 19556343 TI - Inhibition of WNT signaling by G protein-coupled receptor (GPCR) kinase 2 (GRK2). AB - Activation of Wnt signaling pathways causes release and stabilization of the transcription regulator beta-catenin from a destruction complex composed of axin and the adenomatous polyposis coli (APC) protein (canonical signaling pathway). Assembly of this complex is facilitated by a protein-protein interaction between APC and a regulator of G protein signaling (RGS) domain in axin. Because G protein-coupled receptor kinase 2 (GRK2) has a RGS domain that is closely related to the RGS domain in axin, we determined whether GRK2 regulated canonical signaling. We found that GRK2 inhibited Wnt1-induced activation of a reporter construct as well as reduced Wnt3a-dependent stabilization and nuclear translocation of beta-catenin. GRK2 enzymatic activity was required for this negative regulatory effect, and depletion of endogenous GRK2 using small interfering RNA enhanced canonical signaling. GRK2-dependent inhibition of canonical signaling is relevant to osteoblast (OB) biology because overexpression of GRK2 attenuated Wnt/beta-catenin signaling in calvarial OBs. Coimmunoprecipitation studies found that: 1) GRK2 bound APC; 2) The GRK2-APC interaction was promoted by GRK2 enzymatic activity; and 3) Deletion of the RGS domain in GRK2 prevented both the GRK2-APC interaction and GRK2-dependent inhibition of canonical signaling. These data suggest that: 1) GRK2 negatively regulates Wnt signaling; 2) GRK2-dependent inhibition of canonical signaling requires a protein-protein interaction between the RGS domain in GRK2 and APC; and 3) Enzymatic activity promotes the GRK2-APC interaction and is required for the negative regulatory effect on canonical signaling. We speculate that inhibiting GRK2 activity in bone-forming OBs might be a useful therapeutic strategy for increasing bone mass. PMID- 19556344 TI - FGF-2 stimulation of RANK ligand expression in Paget's disease of bone. AB - Receptor activator for nuclear factor-kappaB ligand (RANKL), a critical osteoclastogenic factor expressed in marrow stromal/preosteoblast cells is up regulated in Paget's disease of bone (PDB). We previously demonstrated that heat shock factor-2 (HSF-2) is a downstream target of fibroblast growth factor-2 (FGF 2) signaling to induce RANKL expression in bone marrow stromal/preosteoblast cells. In this study, we identified a 2.5-fold increase in serum FGF-2 levels in patients (n = 8) with PDB compared with normal subjects (n = 10). We showed that HSF-2 co-immunoprecipitates with heat-shock protein-27 (HSP-27) and that FGF-2 stimulation significantly increased phospho-HSP-27 levels in marrow stromal cells. Confocal microscopy revealed HSF-2 colocalization with HSP-27 in unstimulated cells and HSF-2 nuclear translocation upon FGF-2 stimulation. We further show that FGF-2 stimulation significantly increased the levels of phosphorylated signal transducers and activators of the transcription (p-STAT-1) in these cells. Western blot analysis confirmed that small interfering RNA suppression of STAT-1 significantly decreased (3.2-fold) RANKL expression and promoter activity in FGF-2-stimulated cells. Chromatin immunoprecipitation assay revealed STAT-1 binding to a putative motif located far upstream (-8 kb) in the hRANKL gene promoter region. These results suggest STAT-1 is a downstream effector of FGF-2 signaling and that elevated levels of FGF-2 stimulates RANKL expression in PDB. PMID- 19556345 TI - Enhancing the apoptotic potential of insulin-like growth factor-binding protein-3 in prostate cancer by modulation of CK2 phosphorylation. AB - IGF-binding protein 3 (IGFBP-3) promotes apoptosis by both IGF-dependent and independent mechanisms. We have previously reported that phosphorylation of IGFBP 3 (S156) by DNA-dependent protein kinase enhances its nuclear accumulation and is essential for its ability to interact with retinoid X receptor-alpha and induce apoptosis in cultured prostate cancer cells. Using specific chemical inhibitors and small interfering RNA, we demonstrate that preventing casein kinase 2 (CK2) activation enhanced the apoptotic potential of IGFBP-3. We mapped potential CK2 phosphosphorylation sites in IGFBP-3 to S167 and S175 and identified that wild type IGFBP-3- and IGFBP-3-S175A-induced apoptosis to a comparable extent. In contrast, IGFBP-3-S167A was far more potently apoptosis inducing due to inability to undergo CK2 phosphorylation. Pretreatment of 22RV1 cells with IGFBP-3 small interfering RNA also limits the ability of high doses of CK2 inhibitor to induce apoptosis. These effects can be reversed by the addition of exogenous IGFBP-3 protein, suggesting reciprocal regulation of cell survival and apoptosis by IGFBP 3 and CK2. These studies reveal multisite phosphorylation of IGFBP-3 that both positively and negatively regulate its apoptotic potential. Understanding such intrinsic regulation of IGFBP-3 action may enhance the development of potential cancer therapies. PMID- 19556346 TI - The 26-amino acid beta-motif of the Pit-1beta transcription factor is a dominant and independent repressor domain. AB - The POU-homeodomain transcription factor Pit-1 governs the pituitary cell specific expression of Pit-1, GH, prolactin (PRL), and TSHbeta genes. Alternative splicing generates Pit-1beta, which contains a 26-amino acid beta-domain inserted at amino acid 48, in the middle of the Pit-1 transcription activation domain (TAD). Pit-1beta represses GH, PRL, and TSHbeta promoters in a pituitary-specific manner, because Pit-1beta activates these same promoters in HeLa nonpituitary cells. Here we comprehensively analyze the role of beta-domain sequence, position, and context, to elucidate the mechanism of beta-dependent repression. Repositioning the beta-motif to the Pit-1 amino terminus, hinge, linker, and carboxyl terminus did not affect its ability to repress basal rat (r) PRL promoter activity in GH4 pituitary cells, but all lost the ability to repress Ras induced rPRL promoter activity. To determine whether beta-domain repression is independent of Pit-1 protein and DNA binding sites, we generated Gal4-Pit-1TAD, Gal4-Pit-1betaTAD, and Gal4-beta-domain fusions and demonstrated that the beta motif is sufficient to actively repress VP16-mediated transcription of a heterologous promoter. Moreover, beta-domain point mutants had the same effect whether fused to Gal4 or within the context of intact Pit-1beta. Surprisingly, Gal4-beta repression lost histone deacetylase sensitivity and pituitary specificity. Taken together, these results reveal that the beta-motif is a context-independent, modular, transferable, and dominant repressor domain, yet the beta-domain repressor activity within Pit-1beta contains cell type, promoter, and Pit-1 protein context dependence. PMID- 19556347 TI - Comparative evolutionary analysis of the major structural subunit of Vibrio vulnificus type IV pili. AB - Type IV pili contribute to virulence in Vibrio vulnificus, the bacterium responsible for the majority of fatal seafood-related infections. Here, we performed within- and between-species evolutionary analysis of the gene that encodes the major structural subunit of the pilus, pilA, by comparing it with pilD and gyrB, the genes encoding the type IV prepilin peptidase and beta subunit of DNA gyrase, respectively. Although the diversity in pilD and gyrB is similar to each other and likely to have accumulated after speciation of V. vulnificus, pilA is several times more diverse at both nonsynonymous and synonymous levels. Also, in contrast to pilD and gyrB, there are virtually unrestricted and highly localized horizontal movements of pilA alleles between the major phylogenetic groups of V. vulnificus. The frequent movement of pilA involves homologous recombination of the entire gene with no evidence for intragenic recombination between the alleles. We propose that pilA allelic diversity and horizontal movement is maintained in the population by both diversifying and frequency dependent selection most likely to escape shellfish innate immunity defense or lytic phages. Other possibilities leading to such selection dynamics of V. vulnificus pilA could involve adaptation to diverse host populations or within host compartments, or natural DNA uptake and transformation. We show that the history of nucleotide diversification in pilA predates V. vulnificus speciation and this diversification started at or before the time of the last common ancestor for V. vulnificus, Vibrio parahaemolyticus, and Vibrio cholerae. At the same time, it appears that within the various pilA groups of V. vulnificus, there is no positive selection for structural mutations and consequently no evidence for source-sink selection. In contrast, pilD has accumulated a number of apparently adaptive mutations in the regions encoding the membrane-spanning portions of the prepilin peptidase, possibly affecting fimbrial expression and/or function, and is being subjected to source-sink selection dynamics. PMID- 19556349 TI - Definition of hypertension for the "old-old". PMID- 19556348 TI - Regional response differences across the human amygdaloid complex during social conditioning. AB - The amygdala is consistently implicated in biologically relevant learning tasks such as Pavlovian conditioning. In humans, the ability to identify individual faces based on the social outcomes they have predicted in the past constitutes a critical form of associative learning that can be likened to "social conditioning." To capture such learning in a laboratory setting, participants learned about faces that predicted negative, positive, or neutral social outcomes. Participants reported liking or disliking the faces in accordance with their learned social value. During acquisition, we observed differential functional magnetic resonance imaging activation across the human amygdaloid complex consistent with previous lesion, electrophysiological, and functional neuroimaging data. A region of the medial ventral amygdala and a region of the dorsal amygdala/substantia innominata showed signal increases to both Negative and Positive faces, whereas a lateral ventral region displayed a linear representation of the valence of faces such that Negative > Positive > Neutral. This lateral ventral locus also differed from the dorsal and medial loci in that the magnitude of these responses was more resistant to habituation. These findings document a role for the human amygdala in social learning and reveal coarse regional dissociations in amygdala activity that are consistent with previous human and nonhuman animal data. PMID- 19556351 TI - Second branchial cleft cyst: not!! PMID- 19556350 TI - Normative apparent diffusion coefficient values in the developing fetal brain. AB - BACKGROUND AND PURPOSE: Previous studies of diffusion-weighted imaging (DWI) in fetuses are limited. Because of the need for normative data for comparison with young fetuses and preterm neonates with suspected brain abnormalities, we studied apparent diffusion coefficient (ADC) values in a population of singleton, nonsedated, healthy fetuses. MATERIALS AND METHODS: DWI was performed in 28 singleton nonsedated fetuses with normal or questionably abnormal results on sonography and normal fetal MR imaging results; 10 fetuses also had a second fetal MR imaging, which included DWI. ADC values in the periatrial white matter (WM), frontal WM, thalamus, basal ganglia, cerebellum, and pons were plotted against gestational age and analyzed with linear regression. We compared mean ADC in different regions using the Tukey Honestly Significant Difference test. We also compared rates of decline in ADC with increasing gestational age across different areas by using the t test with multiple comparisons correction. Neurodevelopmental outcome was assessed. RESULTS: Median gestational age was 24.28 weeks (range, 21-33.43 weeks). Results of all fetal MR imaging examinations were normal, including 1 fetus with a normal variant of a cavum velum interpositum. ADC values were highest in the frontal and periatrial WM and lowest in the thalamus and pons. ADC declined with increasing gestational age in periatrial WM (P = .0003), thalamus (P < .0001), basal ganglia (P = .0035), cerebellum (P < .0001), and pons (P = .024). Frontal WM ADC did not significantly change with gestational age. ADC declined fastest in the cerebellum, followed by the thalamus. CONCLUSIONS: Regional differences in nonsedated fetal ADC values and their evolution with gestational age likely reflect differences in brain maturation and are similar to published data in premature neonates. PMID- 19556352 TI - Steroid-responsive large vessel vasculitis: application of whole-brain 320 detector row dynamic volume CT angiography and perfusion. AB - SUMMARY: A patient with suspected giant cell arteritis and prior negative findings on superficial temporal artery biopsy was evaluated with 320-detector row CT angiography (CTA) and whole-brain perfusion. Corticosteroid treatment was initiated on the basis of CT angiography findings of arteritis and a cortical perfusion deficit. The patient's symptoms and perfusion imaging findings resolved following therapy. Whole-brain CTA and imaging was helpful in the diagnosis and monitoring this patient with suspected vasculitis. PMID- 19556353 TI - Neuroradiologic manifestations of Loeys-Dietz syndrome type 1. AB - BACKGROUND AND PURPOSE: Loeys-Dietz syndrome (LDS) is a recently described entity that has the triad of arterial tortuosity and aneurysms, hypertelorism, and bifid uvula or cleft palate. Its neuroradiologic manifestations have not been well delineated. We sought to describe the neuroradiologic features of LDS and to assess the manifestations that would warrant follow-up imaging. MATERIALS AND METHODS: Two neuroradiologists retrospectively reviewed CT angiography (CTA), MR imaging, and plain film studies related to the head and neck in 25 patients ranging from 1 to 55 years of age, all of whom had positive genetic testing and clinical characteristics of LDS. Arterial tortuosity was evaluated by subjective assessment of 2D and 3D volumetric CTA and MR angiography data. Craniosynostosis and spinal manifestations were assessed by using plain films and CT images. MR images mostly of the head were reviewed for associated findings such as hydrocephalus, Chiari malformation, etc. Clinical manifestations were collated from the electronic patient record. RESULTS: All patients had extreme arterial tortuosity, which is characteristic of this syndrome. Thirteen patients had scoliosis, 12 had craniosynostosis, 8 had intracranial aneurysms, 6 had spinal instability, 3 had dissections of the carotid and vertebrobasilar arteries, 3 had hydrocephalus, 4 had dural ectasia, 2 had a Chiari malformation, and 1 had intracranial hemorrhage as a complication of vascular dissection. CONCLUSIONS: Significant neuroradiologic manifestations are associated with LDS, predominantly arterial tortuosity. Most of the patients in this series were young and, therefore, may require serial CTA monitoring for development of intra- and extracranial dissections and aneurysms, on the basis of the fact that most of the patients with pseudoaneurysms and dissection were older at the time of imaging. Other findings of LDS such as craniosynostosis, Chiari malformation, and spinal instability may also need to be addressed. PMID- 19556354 TI - Prediction of functional outcome in acute cerebral hemorrhage using diffusion tensor imaging at 3T: a prospective study. AB - BACKGROUND AND PURPOSE: Early evaluation of the pyramidal tract is a prerequisite in patients with intracerebral hemorrhage (ICH) in order to decide the optimal treatment or to assess appropriate rehabilitation. The aim of this study was to evaluate and predict the neuromotor and functional outcome of an ICH by using diffusion tensor imaging (DTI) in the acute phase. MATERIALS AND METHODS: Eighteen patients with a hemiparetic supratentorial ICH were prospectively studied with DTI within 2 days after onset. A region-of-interest-based analysis was performed for the fractional anisotropy (FA) of the pyramidal tract in the cerebral peduncles. The degree of paresis was assessed at day 0 and day 28 by paresis grading (PG). The functional outcome was evaluated by the modified Rankin Scale (mRS). RESULTS: The FA in the affected side was significantly lower compared with that of the unaffected side (P = .001) with the mean diffusivity remaining unchanged (P = .50). The ratio of the FA (rFA) in the affected side to the unaffected side was significantly correlated with the PG at day 0 and 28 and the mRS score at day 28 (P = .002, r = -0.674; P < .001, r = -0.767; and P = .002, r = -0.676). The rFA for the good and poor outcomes based on the PG was significantly different (P < .001). The cutoff point of the rFA for the good and poor outcomes was set at 0.85 (sensitivity, 100%, specificity, 100%). CONCLUSIONS: We conclude that DTI can evaluate the motor deficit quantitatively and may predict the functional outcome in patients with an ICH who were scanned within 2 days after the ICH onset. PMID- 19556355 TI - Common variants in KCNQ1 are associated with type 2 diabetes and impaired fasting glucose in a Chinese Han population. AB - Common variants in KCNQ1 have recently been reported to be associated with type 2 diabetes in East Asians. We aimed to examine whether these common variants (rs2074196, rs2237892, rs2237895 and rs2237897) were also associated with type 2 diabetes in a population-based cohort of 3210 Chinese Hans and to explore the underlying mechanisms. The SNPs rs2237892, rs2237895 and rs2237897 were significantly associated with type 2 diabetes (OR: 1.33-1.36, P 8.0 in which catheter encrustation was inhibited. It is suggested that this dietary strategy will be an effective means of controlling catheter encrustation, whichever bacterial species is causing the problem. PMID- 19556374 TI - Let's not ignore the statistics. PMID- 19556376 TI - Hepcidin for clinicians. AB - Despite the use of erythropoiesis-stimulating agents (ESAs), the anemia of chronic kidney disease (CKD) can be resistant to therapy. Both absolute and functional iron deficiency along with inflammation can contribute to ESA resistance and can be difficult to identify with current-day markers of iron storage. Hepcidin, a small peptide produced by the liver, is a recently discovered key regulator of iron homeostasis. Via regulation of ferroportin, hepcidin inhibits intestinal iron absorption and iron release from macrophages and hepatocytes. Because of its renal elimination and regulation by inflammation, it is possible that progressive renal insufficiency leads to altered hepcidin metabolism, subsequently affecting enteric absorption of iron and the availability of iron stores. Thus, hepcidin likely plays a major role in the anemia of CKD as well as ESA resistance. This article discusses the biologic actions and regulation of hepcidin along with reviewing studies of hepcidin in CKD. PMID- 19556377 TI - Vitamin D and parathyroid hormone in general populations: understandings in 2009 and applications to chronic kidney disease. AB - Vitamin D is now recognized as an important prohormone in health and disease. Its role in immunoregulation and cardiovascular and bone health has become topical in the lay press and the medical press in the past 5 yr. The target audience for this review includes the interested clinician and researchers. The prevalence of chronic kidney disease in the general population has further increased the interest and perhaps the applicability of findings of population studies. The basic physiology of vitamin D and receptor activation and biologic importance is reviewed, as well as various vitamin D analogues and nomenclature. Issues related to measurement of vitamin D and parathyroid hormone have the potential to complicate the clinical use of these tests and should be understood by all clinicians so as to ensure informed decision making and stimulate interest in participation in clinical trials. The epidemiology of vitamin D deficiency and supplementation in association with health status and disease status is reviewed, and issues related to association versus causation are highlighted. Some recommendations for pragmatic approaches and study design are suggested. PMID- 19556378 TI - Hypertension in pediatric long-term hemodialysis patients in the United States. AB - BACKGROUND AND OBJECTIVES: Data are limited regarding BP distribution and the prevalence of hypertension in pediatric long-term dialysis patients. This study aimed to examine BP distribution in U.S. pediatric long-term hemodialysis patients. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This cross-sectional study of all U.S. pediatric (aged 0-< 18 yr, n = 624) long-term hemodialysis patients was performed as part of the Centers for Medicare & Medicaid Services End-Stage Renal Disease (ESRD) Clinical Performance Measures Project. BP and clinical information were collected monthly in October, November, and December 2001. Hypertension was defined as the mean of pre- and postdialysis systolic or diastolic BP above the 95th percentile for age, height, and sex, or antihypertensive medication use. Results were calculated by age, sex, race, ethnicity, ESRD duration, body mass index percentile, primary cause of ESRD, and laboratory data. RESULTS: Hypertension was present in 79% of patients; 62% used antihypertensive medication. Five percent of patients were prehypertensive (mean BP at 90th to 95th percentile). Hypertension was uncontrolled in 74% of treated patients. Characteristics associated with hypertension included acquired kidney disease, shorter duration of ESRD, and lower mean hemoglobin and calcium values. Characteristics associated with uncontrolled hypertension were younger age and shorter duration of ESRD. CONCLUSIONS: Hypertension is common in U.S. pediatric long-term hemodialysis patients, uncontrolled in 74% of treated patients, and untreated in 21% of hypertensive patients. It is concluded that a more aggressive approach to treatment of hypertension is warranted in pediatric long-term hemodialysis patients. PMID- 19556379 TI - Complement inhibitor eculizumab in atypical hemolytic uremic syndrome. AB - BACKGROUND AND OBJECTIVES: Atypical hemolytic uremic syndrome (aHUS) is associated with a congenital or acquired dysregulation of the complement alternative pathway that leads to continuous complement activation on host cells causing inflammation and damage. Eculizumab, a humanized mAb against complement protein C5, inhibits activation of the terminal complement pathway. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We report an adolescent with relapsing unclassified aHUS. On admission, a high plasma creatinine level indicated a poor prognosis, and hemodialysis had to be started. Plasma exchanges were initially effective against the microangiopathic hemolytic activity and allowed a temporary improvement of renal function with termination of hemodialysis after 7 wk. Subsequently, plasma exchanges (three times per week) failed to prevent ongoing aHUS activity and progressive renal failure. After 12 wk, aHUS treatment was switched to eculizumab. RESULTS: Eculizumab was effective in terminating the microangiopathic hemolytic process in two aHUS relapses; however, after normalization of complement activity, aHUS recurred and ultimately led to anuric end-stage renal failure. CONCLUSIONS: In this patient, complement inhibition by eculizumab temporarily terminated the microangiopathic hemolytic activity. Nevertheless, renal damage as a result of preceding and subsequent aHUS activity resulted in end-stage renal failure; therefore, therapeutic success may depend on early administration of eculizumab. The optimal duration of treatment may be variable and remains to be determined. PMID- 19556380 TI - Hydration status does not influence peritoneal equilibration test ultrafiltration volumes. AB - BACKGROUND AND OBJECTIVES: The peritoneal equilibration test (PET) was developed some 25 yr ago and has been used to help prescribe peritoneal dialysis. However, PET is affected by several factors, including diabetes and inflammation. It was speculated that extracellular fluid overload would increase PET ultrafiltration volumes, and therefore the usefulness of the PET in routine clinical practice was audited. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Data from 211 consecutive patients attending a university teaching hospital for a standard PET who had multifrequency bioimpedance performance were analyzed to determine which factors affected net PET ultrafiltration volumes. RESULTS: Net PET ultrafiltration volume was independent of gender, age, diabetes, residual renal function, peritoneal dialysis prescriptions (modes and dialysates), extracellular fluid volume, or C reactive protein (CRP). There was an inverse regression with serum albumin and sodium on multiple logistical regression analysis (F = 13.4, P < 0.001 and F = 10.1, P = 0.001, respectively) and a positive regression with 24-h net peritoneal ultrafiltration volumes (F = 15.5, P < 0.001). As expected, there was a strong correlation with net sodium losses (r = 0.99, P < 0001). CONCLUSIONS: It was found that PET test ultrafiltration volume in routine clinical practice was not affected by CRP, hyperglycemia, or extracellular fluid volume overload. Ultrafiltration volumes were increased in those patients with reduced serum sodium and albumin, most likely because of inflammation and protein malnutrition. PMID- 19556382 TI - Thyroglobulin as a marker of iodine nutrition status in the general population. AB - OBJECTIVE: The iodine status of a population is traditionally evaluated by either urinary iodine (UI) excretion or by some measure of thyroid volume and the prevalence of goitre. In this prospective study of a mandatory iodization programme, we aimed to evaluate serum thyroglobulin (Tg) as a marker of iodine status in the population. METHODS: Two identical cross-sectional studies were performed before (1997-1998, n=4649) and after (2004-2005, n=3570) the initiation of the Danish iodization programme in two areas with mild and moderate iodine deficiency. Serum Tg was measured from blood samples. Thyroid volume was measured by ultrasonography. RESULTS: Before iodization, the median serum Tg was considerably higher in moderate than in mild iodine deficiency. Iodization led to a lower serum Tg in all examined age groups. The marked pre-iodization difference in Tg level between the regions was eliminated. The prevalence of Tg above the suggested reference limit (40 microg/l) decreased from 11.3 to 3.7% (P<0.0001). Using bootstrapping, we demonstrated a higher efficacy of Tg than of thyroid volume to show a difference between pre- and post-iodization values. CONCLUSION: We found serum Tg to be a suitable marker of iodine nutrition status in the population. The results may suggest that the Danish iodization programme has led to a sufficient iodine intake, even if the median UI excretion is still marginally low according to WHO criteria. PMID- 19556381 TI - Contrast-induced nephropathy and long-term adverse events: cause and effect? AB - BACKGROUND AND OBJECTIVES: The relationship of contrast-induced nephropathy (CIN) to long-term adverse events (AEs) is controversial. Although an association with AEs has been previously reported, it is unclear whether CIN is causally related to these AEs. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We obtained long term (> or =1 yr) follow-up on 294 patients who participated in a randomized, double-blind comparison of two prevention strategies for CIN (iopamidol versus iodixanol). A difference in the incidence of AEs between patients who had developed CIN and those who had not was performed using a chi(2) test and Poisson regression analysis. A similar statistical approach was used for the differences in AEs between those who received iopamidol or iodixanol. Multiple definitions of CIN were used to strengthen and validate the results and conclusions. RESULTS: The rate of long-term AEs was higher in individuals with CIN (all definitions of CIN). After adjustment for baseline comorbidities and risk factors, the adjusted incidence rate ratio for AEs was twice as high in those with CIN. Randomization to iopamidol reduced both the incidence of CIN and AEs. CONCLUSIONS: The parallel decrease in the incidence of CIN and AEs in one arm of this randomized trial supports a causal role for CIN. PMID- 19556383 TI - Children's physical activity, TV watching and obesity in Cyprus: the CYKIDS study. AB - BACKGROUND: Even though there is a severe obesity problem in Cyprus, information about the contribution of predisposing lifestyle factors is limited. Our aim was to investigate the relationship between physical activity (PA), sedentary behaviour and various obesity indices [i.e. body mass index (BMI), waist circumference (WC), percentage of body fat (BF%) and 'total & abdominal obesity' (TAO)]. METHODS: A national cross-sectional study of 1140 children (mean age = 10.7 +/- 0.98 years) selected by multistage sampling in Cyprus was conducted during 2004-05. Children completed a 32-item, semi-quantitative PA questionnaire, which assessed organized and free-time PA and sedentary behaviours. Weight, height and WC were collected from a random sub-sample of 622 children and obesity was defined by IOTF criteria. Body fat percentage was calculated, and TAO status was computed based on obesity status and WC [i.e. (i) both BMI/WC, (ii) either BMI/WC abnormal and (iii) both BMI/WC abnormal]. Linear and logistic regression analyses with obesity indices as dependent variables were applied after adjusting for several potential confounders. RESULTS: Only variables describing sedentary behaviours were retained in the final regression models in both boys and girls. Girls who spent >or=4 h/day on TV and DVD watching were almost three times more likely to be overweight or obese [OR = 2.84 (95% CI 1.08-7.47)], three times more likely to have WC >or=75th percentile [OR = 3.25 (95% CI 1.06-9.98)] and 3.5 times more likely to have >or=30% body fat [OR = 3.63 (95% CI 1.01-12.98)], while in boys, even though the same variable was retained in almost all final models, it did not reach statistical significance. CONCLUSION: Sedentary behaviours such as TV watching may be more important predictors of children's various obesity indices than PA behaviours. Interventions targeting sedentary behaviours, such as TV watching, may help in the prevention and treatment of obesity among Cypriot children. PMID- 19556384 TI - Underappreciated role of binge drinking in the risk of lung cancer. PMID- 19556385 TI - Deaths in German police custody. AB - BACKGROUND: According to the ministries of the interior of all German federal states, a total of 128 persons died in police custody between 1993 and 2003. METHODS: An inquiry to the forensic university institutes and regional court doctors showed that, within this period, post mortems were conducted in 75 of these cases. We were able to include 60 of these 75 cases in our evaluation and deficiency analysis, which were carried out under assessment of the post mortem records and partly by making use of the subsequent medical expert reports and the results of criminal investigation. RESULTS: The most frequent cause of death was acute alcoholic intoxication, followed by cerebrocranial traumata, internal diseases and fatal poisonings with medical or illegal drugs. In 23 cases, the person taken into custody had been seen by a physician in order to determine the person's fitness for detention in custody. Of these 23 cases, 15 (65.2%) revealed deficiencies in various areas of medical activity. Police officers had made mistakes in 33 of the 60 cases (55.0%), mainly because they failed to seek medical assistance or did not monitor the person with sufficient frequency. Our retrospective study however showed that in 16 cases (26.6%) death was very probably not preventable even if the person had received all necessary care. CONCLUSIONS: Our study gives recommendations on how to improve various measures to increase the quality of medical attention given to persons taken into police custody. PMID- 19556386 TI - Intent to "teach" and pay to play in osteopathic medical education. PMID- 19556387 TI - Personality types and performance on aptitude and achievement tests: implications for osteopathic medical education. AB - CONTEXT: Several studies have shown that the personality types of medical and dental students affect performance on aptitude and achievement examinations. However, such studies are lacking in osteopathic medical literature. OBJECTIVE: To determine if personality type is associated with performance on aptitude and achievement tests taken by osteopathic medical students. METHODS: The Myers Briggs Type Indicator (MBTI) was used to determine the mental-function pairs sensing-thinking, intuition-thinking, sensing-feeling, or intuition-feeling-of osteopathic medical students at Midwestern University/Chicago College of Osteopathic Medicine in Downers Grove, Ill. Results were analyzed with participants' scores on the Medical College Admissions Test and the Comprehensive Osteopathic Medical Licensing Examination-USA (COMLEX-USA) Level 1. RESULTS: A total of 295 osteopathic medical students completed the MBTI, but 32 (11%) were excluded because they did not meet the study criteria. Among the remaining 263 participants, no personality types were associated with high or low scores on the Medical College Admissions Test (P=.229). However, participants in the intuition feeling group had statistically significant lower scores on COMLEX-USA Level 1 (P=.002). CONCLUSION: The differences in scores obtained on COMLEX-USA Level 1 were statistically significant when students were identified by personality type. This finding suggests that using the MBTI during training could enhance learning and improve academic performance in osteopathic medical schools. PMID- 19556388 TI - Measuring awareness, interest, and involvement in the osteopathic community through board certification: a survey of DO residents in ACGME-accredited training programs. AB - Currently, close to 50% of osteopathic medical graduates receive residency training from programs accredited by the Accreditation Council for Graduate Medical Education (ACGME) rather than those approved by the American Osteopathic Association (AOA). As a result, leaders within the osteopathic medical profession have expressed ongoing concerns about the viability of the profession's distinct osteopathic identity. Using a one-page, 12-item survey, the authors queried ACGME trained family practice residents (N=1354) regarding their interest in formal membership, continuing medical education activities, and specialty board certification options within the osteopathic medical profession. Four hundred twenty-six completed surveys were returned and usable for analysis for an overall response rate of 31.4%. A majority of survey participants indicated an interest "in continuing [their] osteopathic skills and training during residency" (376 [88.5%]), membership in osteopathic organizations and participating in continuing medical education programs (325 [77.2%]), and completing the American Osteopathic Board of Family Physicians certification examination (267 [63.7%]). Unfortunately, actual involvement may be limited by lack of communication or understanding, as in the case of lack of awareness regarding eligibility criteria for AOA board certification (311 [74.2%]). A variety of recommendations are offered to osteopathic organizations to improve involvement in and commitment to the profession among ACGME-trained DOs. PMID- 19556389 TI - The role of the adipocytokines adiponectin and leptin in migraine. AB - Although it has long been known that fasting or the consumption of certain foods can trigger headaches, abdominal and total body obesity have only recently been linked to migraine. Several adipocytokines appear to play an integral role in feeding and obesity--and have also been linked to pain. Among these proteins are adiponectin and leptin. The author reviews the regulation of adipose tissue and feeding and provides an in-depth examination of adiponectin and leptin and their association with migraine. PMID- 19556390 TI - Perception of osteopathic medicine among allopathic physicians in the deep central southern United States. AB - CONTEXT: Relatively few osteopathic physicians (DOs) practice in the deep central southern geographic region as compared to other parts of the United States. OBJECTIVE: To assess the potential effects of underrepresentation on the perception of osteopathic medicine among allopathic physicians (MDs) in this geographic region. METHODS: A 20-item, Likert scale survey was designed to evaluate participant perceptions of the osteopathic medical profession. The instrument was mailed to 468 MDs who reside and practice in the deep central southern region of the United States. RESULTS: One hundred seven individuals completed the survey for a response rate of 22.9%. Although the majority of respondents (71 [66.3%]) recognized the distinctiveness of the osteopathic medical profession, they were not necessarily able to articulate these differences clearly through their responses to other survey items. In addition, survey participants expressed was a belief that residency training programs accredited by the Accreditation Council for Graduate Medical Education were more beneficial to osteopathic medical graduates than those approved by the American Osteopathic Association (94 [87.8%]). Finally, there was a perception that research efforts supporting "the scientific basis of osteopathic medicine" were inadequate (53 [49.5%]). The one demographic factor that had a statistically significant (P<.001) positive impact on perceptions of osteopathic medicine among these MDs was previous contact with DOs. CONCLUSION: Underrepresentation of DOs in the deep central southern region of the United States appears to have an impact on the perception of osteopathic medicine among MDs insofar as direct or indirect contact with osteopathic physicians led to improved perceptions among this cohort. PMID- 19556391 TI - Methotrexate-induced Hodgkin disease in a patient with systemic lupus erythematosus. AB - Methotrexate sodium use in the management of various immunologic disorders has increased--as have the number of reported adverse effects associated with this therapy. While methotrexate has helped combat various autoimmune and cancerous disorders, the paradoxical risk of causing an often fatal malignancy may still occur as a result of the drug's effect on suppressing immune function. We present a case of methotrexate-induced Hodgkin disease in a 48-year-old man with a history of systemic lupus erythematosus (SLE). Discontinuation of methotrexate facilitated Hodgkin disease reversal. In addition, we review other lymphoproliferative hematologic malignancies caused by methotrexate. PMID- 19556392 TI - Leu27IGF2 plays an opposite role to IGF1 to induce H9c2 cardiomyoblast cell apoptosis via Galphaq signaling. AB - This study examines the role of IGF2/mannose 6-phosphate receptor (IGF2R) signaling in the signaling transduction regulation and cell apoptosis in H9c2 cardiomyoblast cells. However, it is difficult to recognize the distinct activation of IGF2 signaling without interfacing with IGFI receptor (IGF1R) after exposure to IGF2. Leu27IGF2, an analog of IGF2 that interacts selectively with the IGF2R, was used to specifically activate IGF2R signaling in this study. DNA fragmentation and TUNEL assay revealed that in contrast to IGF1 treatment preventing angiotensin II and AG1024-induced cell apoptosis, Leu27IGF2 appears to synergistically increase apoptosis in those cells. We further found cell apoptosis induction and an increase in the active form of caspase 3 in the treatment of cells with Leu27IGF2, but not IGF1. To detect the interaction between IGF2R and Galphaq using the immunoprecipitation assay, we found that IGF2R could directly interact with Galphaq and after treatment with Leu27IGF2 the binding ability of Galphaq to IGF2R had increased. This sequentially resulted in the phosphorylation of phospholipase C-beta, a key downstream modulator of Galphaq, on serine 537. Moreover, disruption of the Galphaq protein by small interferon RNA reduced the cell apoptosis induced by Leu27IGF2. Our findings demonstrate that IGF2R activation appears to induce cell apoptosis via Galphaq deriving signaling cascades and its effect is completely different from IGF1R survival signaling. PMID- 19556394 TI - Changes in social participation and volunteer activity among recently widowed older adults. AB - PURPOSE: Widowhood eliminates a key source of support that may trigger greater involvement in social activities and volunteer participation, which are related to better late-life health and functioning. We reexamine and build upon 2 recent studies exploring recent widowhood and social participation. Using different data, we perform a quasi-replication of Utz, Carr, Nesse, and Wortman's (2002; "The effect of widowhood on older adults' social participation: An evaluation of activity, disengagement, and continuity theories," The Gerontologist, 42, 522 533) study and employ different analytic strategies to Li's (2007; "Recovering from spousal bereavement in later life: Does volunteer participation play a role?" Journal of Gerontology Series B: Psychological Sciences and Social Sciences, 62, S257-S266) study. DESIGN AND METHODS: A synthetic cohort of recently widowed individuals aged 60 years and older (n = 228) was compared with random, non-widowed older adult controls (n = 228) across 3 waves of Americans' Changing Lives data. Generalized estimating equations were used to assess the impact of widowhood on levels of social participation and formal and informal volunteerism, controlling for social, economic, demographic, and psychological factors. RESULTS: Similar to Utz and colleagues, we found that widowhood was positively related to informal social participation, net of other effects, but did not reproduce this finding for formal social participation. Unlike Li, we did not find a significant relationship between widowhood and formal or informal volunteerism. Controlling for prior participation, widowhood remained significantly related to informal and formal social participation. IMPLICATIONS: Older adults increase their reliance on sources of other social support following spousal loss but do not change their volunteer activities. This suggests that continuity of volunteer engagement and enhanced social participation are important following widowhood. Given their positive associations with late-life well-being, efforts to help older widows and widowers increase their social participation and maintain established patterns of volunteerism following spousal loss are warranted. PMID- 19556393 TI - Silver alerts and the problem of missing adults with dementia. AB - In the months following the introduction of the National AMBER (America's Missing: Broadcast Emergency Response) Alert plan used to locate missing and abducted children, Silver Alert programs began to emerge. These programs use the same infrastructure and approach to find a different missing population, cognitively impaired older adults. By late 2008, 17 states had enacted Silver Alert policies, and several more planned to take advantage of National Silver Alert grant funding to initiate policies in 2009. To date, however, no research has examined the efficacy of such programs, which have widely varying parameters and criteria to initiate the alerts. In this study, we empirically examine the 17 existing state Silver Alert and related policies. The analysis includes an examination of the varieties of programs: dementia related and AMBER extension, the dates of enactment, the criteria for activation, and the process of activation. We conclude with two salient questions that emerged from the analysis. We examine these questions and make recommendations for future research, including examining whether Silver Alerts are an appropriate response to address the problem of missing adults with dementia or cognitive impairments and examining the costs and benefits of the programs including determining how best to balance efforts to keep cognitively impaired elders safe while keeping their basic human rights of autonomy and empowerment intact. PMID- 19556397 TI - Physical activity: not just for quantitative researchers. PMID- 19556395 TI - Towards temperature-dependent coarse-grained potentials of side-chain interactions for protein folding simulations. I: molecular dynamics study of a pair of methane molecules in water at various temperatures. AB - By means of molecular dynamics simulations of a pair of methane molecules in a TIP3P periodic water box with the NVT scheme at six temperatures and, additionally, the NPT scheme at three temperatures ranging from T = 283 to 373 K, we determined the potential of mean force (PMF) of pairs of interacting methane molecules in water as functions of distance between the methane molecules. The PMFs converge to a single baseline only for r> 11 A at all temperatures. The curves of the dimensionless PMF obtained at different temperatures with the NVT scheme overlap almost perfectly in the region of the contact minimum and still very well in the regions of the desolvation maximum and the solvent-separated minimum, which suggests that the temperature-dependent hydrophobic interaction potentials at constant volume in united-residue force fields can be obtained by scaling the respective dimensionless potentials by RT, R being the universal gas constant. For the dimensionless potentials of mean force obtained with the NPT scheme, the depth of the contact minimum increases, whereas the height of the desolvation maximum and the depth of the solvent-separated minimum decrease with temperature, in agreement with results reported in the literature. PMID- 19556396 TI - Q fever: persistence of antigenic non-viable cell residues of Coxiella burnetii in the host--implications for post Q fever infection fatigue syndrome and other chronic sequelae. AB - BACKGROUND: Our previous studies of persistence of Coxiella burnetii in humans after an initial acute Q fever infection revealed raised, maintained antibody levels and low levels of coxiella genomic DNA at the age of 5 years from onset in Australian patients and at 12 years in patients in the 1989 Birmingham UK Q fever outbreak. Attempts to isolate the coxiella in standard cell culture and susceptible mice by serial passage of PCR positive PBMC and bone marrow were negative. AIM: To retest PCR positive patient samples by more sensitive methods for viable coxiellas and for the coxiella cell components of antigen and specific lipopolysaccharide (LPS). To re-interpret the previous results in the light of the new information. To review the pertinent literature for a concept of an immuno-modulatory complex generated by the current studies. DESIGN: Laboratory case study. METHODS: Stored patient samples were inoculated into SCID mice that were followed for 60 days. Mouse spleen and liver samples were then examined by PCR assay for targets in the COM1 and IS1111a sequences and for antigens by IFA with a polyclonal rabbit antiserum to C. burnetii Phase 1 and a monoclonal antiserum to Phase 1 LPS (details; O. Sukocheva et al., unpublished data). RESULTS: All specimens, including a recently excised heart valve from a Birmingham patient with late developing endocarditis, were infection negative in SCID mice. Dilutions of SCID mouse spleen and liver homogenates titrated in PCR assays were negative at dilutions attained by control mice inoculated with an endpoint dilution of a viable prototype strain of C. burnetii. Sections of the spleens from all specimens showed a complex of coxiella antigen-LPS by IFA. DISCUSSION/REVIEW: We advance a concept of long-term persistence of a non infective, non-biodegraded complex of coxiella cell components with its antigens and specific LPS [so called Immunomodulatory complex (IMC)] associated with traces of genomic DNA that signalled its presence in our earlier studies. The IMC's survival in patients for at least 12 years, and in one patient for 70 years implies a capacity for serial passage in macrophages with effective down regulation of their biodegrading functions. The review assesses the compatibility of the IMC concept in relation to cogent literature on C. burnetii interactions with macrophage and cell-mediated immunity. Some remaining gaps in our knowledge of the organ sites and duration of carriage of viable coxiellas after initial infection are also identified. PMID- 19556398 TI - Using a socioecological approach to examine participation in sport and physical activity among rural adolescent girls. AB - Adolescence is a critical time for developing lifelong healthy behaviors, including active lifestyles. Participation in sport and physical activity, however, declines during adolescence, and few studies have comprehensively identified why, particularly among rural girls. This article identifies a range of independent and interacting factors that influence sport and physical activity participation of rural adolescent girls. The socioecological model of health was used to guide four focus group discussions with Grade 7 girls (n = 34). The results showed that adolescent girls were positively influenced when sports or physical activities were fun, when they involved being with friends, and when they were supported by families and teachers through role modeling and positive feedback. A range of intrapersonal and organizational factors affected perceived self-competence, particularly the coeducational nature of school physical education classes and peer teasing, which supported social comparisons of skill level. In promoting sport and physical activity to rural adolescent girls, focus must be directed on developmentally appropriate activities that are fun, offering opportunities for single-sex classes, and generating cultural changes that encourage noncompetitive and self-referencing activities. PMID- 19556399 TI - The construction of patients' involvement in hospital bedside teaching encounters. AB - Active and sensitively managed patient involvement in medical education is vital in the drive toward the development of tomorrow's doctors' patient-centered professionalism. Bedside teaching encounters (BTE) involve clinicians, medical students, and patients, and comprise a formative and focused activity through which students learn both the "whats" and "hows" of physicianship. We present four case studies from our analysis of six BTEs, drawing on Goffman's dramaturgy theory and broader interactional research. We demonstrate the multiple roles participants play within BTEs, including actor, director, audience, nonperson, and prop. Although patients sometimes participated as team members, even taking the role of director, they were commonly positioned in less active roles: as audience, nonperson, and prop. In this article we discuss critically this commonality across the BTEs: the patient role as passive object at the point at which the serious act of teaching physical examination begins, even when patients actively reject this passive role. PMID- 19556400 TI - Social support and unsolicited advice in a bipolar disorder online forum. AB - How does a newly diagnosed user get inducted into a forum dedicated to people suffering from bipolar disorder? Is their opening message "matched" by the forum's reply? We add to the literature on social support online by using conversation analysis (CA) to explore an apparent contradiction between a new user's first post and forum members' replies with ostensibly unsolicited advice. CA reveals the intimate relation between turns in sequence, an aspect of online communication largely ignored in existing work on social support. Seen from this perspective, giving unsolicited advice, although apparently a "mismatch," turns out to be a consequence of the open design of the new user's initial posting. We speculate that such unsolicited advice might function at the ideological level to induct the new user into the mores of the group, not only in the kind of support it countenances giving, but into the very meaning of bipolarity itself. PMID- 19556402 TI - Elder authority and the situational diagnosis of diarrheal disease as normal infant development in northeast Thailand. AB - Research was conducted in rural Northeast Thailand to understand how mothers perceive children's acute illness episodes, and their resulting illness management strategies. Although diarrheal disease is one of the leading causes of illness among young children in Thailand, interviews revealed that mothers frequently do not classify infantile diarrhea as an "illness." Infantile diarrhea is commonly labeled thai su, a type of diarrhea believed necessary to "lighten the body" so that the infant can attain a new developmental stage, such as sitting up, standing, or walking. Consequently, mothers do not perceive infection related diarrheal illness as common in infants, and therefore do not direct much attention to prevention, nor manage diarrheal cases in a manner consistent with biomedical recommendations. In this article I describe local explanatory models of diarrheal illness, and discuss the role of situational factors and elder authority in determining how illness episodes are assessed and managed. PMID- 19556401 TI - Birth talk in second stage labor. AB - In this secondary analysis of videotape data, we describe birth talk demonstrated by caregivers to women during the second stage of labor. Birth talk is a distinctive verbal register or a set of linguistic features that are used with particular behaviors during specific situations, has a particular communication purpose, and is characterized by distinctive language features. Birth talk is found cross-culturally among speakers of diverse languages. Our findings show that birth talk occurred mainly during contractions and co-occurred with two general styles of caregiving: "directed toward forced bearing down" and "supportive of physiologic bearing down." We also describe talk that occurred during rest periods, which was similar across the two styles. Caregivers' use of language tended to be either procedural (giving directions, instructions) or comfort related (encouraging and supporting). Linguistic features of the talk consisted of utterances of short duration, level pitch patterns with no sudden pitch shifts, and a restricted pitch range. PMID- 19556403 TI - Food that makes you different: the stigma experienced by adolescents with celiac disease. AB - For adolescents with celiac disease (CD), a gluten-free diet (GFD) is crucial for health, but compliance is problematic and noncompliance is common even among those aware of the risks. To better understand their lives with the disease, Swedish CD adolescents were invited to take part in focus group discussions. Data were analyzed for recurrent stigma-related themes across the groups. Adolescents described an awareness of being different from others that was produced by meal appearance and the poor availability of gluten-free food. The GFD often required discussions and special requests, so eating in public had the effect of making an invisible condition visible, and thereby creating a context for felt or enacted stigma. Maintaining invisibility avoided negative consequences of stigma, and other strategies were used to reduce the costs of visibility. The results of the study show that the GFD can produce stigma experiences in adolescence, and that dietary compliance (or lack thereof) can be understood in terms of dealing with GFD concealment and disclosure. PMID- 19556404 TI - In a country as affluent as America, people should be eating: experiences with and perceptions of food insecurity among rural and urban Oregonians. AB - Many factors are associated with food insecurity in the United States. We conducted interviews with 25 low-income and/or food-insecure Oregonians to explore their experiences with food insecurity, the role of social support, and whether these experiences differed based on rural/urban residence. Ill health and unemployment emerged as food-insecurity contributors. Coping strategies cited included use of nutrition assistance programs, alternate food sources, and drawing on social support. The findings suggest that policy and practice efforts should be directed at increasing the human capital of low-income Oregonians and the benefit levels of essential nutrition assistance programs. PMID- 19556405 TI - The incidence of adverse events in Swedish hospitals: a retrospective medical record review study. AB - OBJECTIVES: To estimate the incidence, nature and consequences of adverse events and preventable adverse events in Swedish hospitals. DESIGN: A three-stage structured retrospective medical record review based on the use of 18 screening criteria. SETTING: Twenty-eight Swedish hospitals. Population A representative sample (n = 1967) of the 1.2 million Swedish hospital admissions between October 2003 and September 2004. MAIN OUTCOME MEASURES: Proportion of admissions with adverse events, the proportion of preventable adverse events and the types and consequences of adverse events. RESULTS: In total, 12.3% (n = 241) of the 1967 admissions had adverse events (95% CI, 10.8-13.7), of which 70% (n = 169) were preventable. Fifty-five percent of the preventable events led to impairment or disability, which was resolved during the admission or within 1 month from discharge, another 33% were resolved within 1 year, 9% of the preventable events led to permanent disability and 3% of the adverse events contributed to patient death. Preventable adverse events led to a mean increased length of stay of 6 days. Ten of the 18 screening criteria were sufficient to detect 90% of the preventable adverse events. When extrapolated to the 1.2 million annual admissions, the results correspond to 105,000 preventable adverse events (95% CI, 90,000-120,000) and 630,000 days of hospitalization (95% CI, 430,000-830,000). CONCLUSIONS: This study confirms that preventable adverse events were common, and that they caused extensive human suffering and consumed a significant amount of the available hospital resources. PMID- 19556406 TI - All ICUs are not created equal: evaluating pilot studies performed in different environments. PMID- 19556408 TI - Not-so-trivial pursuit: mechanical ventilation risk reduction. AB - As many as half of critically ill patients require mechanical ventilation. In this article, a program of research focused on reduction of risk associated with mechanical ventilation is reviewed. Airway management practices can have profound effects on outcomes in these patients. How patients are suctioned, types of processes used, effects of suctioning in patients with lung injury, and open versus closed suctioning systems all have been examined to determine best practices. Pneumonia is a common complication of mechanical ventilation (ventilator-associated pneumonia), and use of higher backrest elevations reduces risk of pneumonia, although compliance with such recommendations varies. The studies reviewed here describe backrest elevation practices, factors that affect backrest elevation, and the effect of backrest elevation on ventilator-associated pneumonia. Oral care strategies also have been investigated to determine their effect on ventilator-associated pneumonia. Oral care practices are reported to hold a low care priority, vary widely across care providers, and differ in intubated versus nonintubated patients. However, in several studies, oral applications of chlorhexidine have reduced the occurrence of ventilator associated pneumonia. Although ventilator patients require sedation, sedation is associated with significant risks. The overall goals of sedation are to provide physiological stability, to maintain ventilator synchrony, and to ensure patients' comfort-although methods to evaluate achievement of these goals are limited. Reducing risks associated with mechanical ventilation in critically ill patients is a complex and interdisciplinary process. Our understanding of the risks associated with mechanical ventilation is constantly changing, but care of these patients must be based on the best evidence. PMID- 19556409 TI - Living with depressive symptoms: patients with heart failure. AB - BACKGROUND: Patients with heart failure often experience depressive symptoms that affect health-related quality of life, morbidity, and mortality. Researchers have not described the experience of patients with heart failure living with depressive symptoms. Understanding this experience will help in developing interventions to decrease depressive symptoms. OBJECTIVE: To describe the experience of patients with heart failure living with depressive symptoms. METHODS: This study was conducted by using a qualitative descriptive design. The sample consisted of 10 outpatients (50% female, mean age 63 [SD, 13] years, 70% New York Heart Association class III or IV) with heart failure who were able to describe depressive symptoms. Data were collected via taped, individual, 30- to 60-minute interviews. ATLAS ti (version 5) was used for content analysis. RESULTS: Participants described emotional and somatic symptoms of depression. Negative thinking was present in all participants and reinforced their depressed mood. The participants experienced multiple stressors that worsened depressive symptoms. The overarching strategy for managing depressive symptoms was "taking my mind off of it." Patients managed depressive symptoms by engaging in activities such as exercise and reading, and by using positive thinking, spirituality, and social support. CONCLUSIONS: Patients with heart failure experience symptoms of depression that are similar to those experienced by the general population. Clinicians should assess patients with heart failure for stressors that worsen depressive symptoms. Strategies that researchers and clinicians can use to reduce depressive symptoms in patients with heart failure include engaging patients in activities, positive thinking, and spirituality. Helping patients find enhanced social support may also be important. PMID- 19556411 TI - Nursing care and intracranial pressure monitoring. PMID- 19556410 TI - Oral care interventions and oropharyngeal colonization in children receiving mechanical ventilation. AB - BACKGROUND: Recent progress in identification of oral microorganisms has shown that the oropharynx can be a site of origin for dissemination of pathogenic organisms to distant body sites, such as the lungs. OBJECTIVE: To compare the oropharyngeal microbiological profile, duration of mechanical ventilation, and length of stay in the intensive care unit of children receiving mechanical ventilation who had pharmacological or nonpharmacological oral care. METHODS: A randomized and controlled study was performed in a pediatric intensive unit in Sao Paulo, Brazil. A total of 56 children were randomly assigned to an experimental group (n=27, 48%) that received oral care with use of 0.12% chlorhexidine digluconate or a control group (n=29, 52%) that received oral care without an antiseptic. Oropharyngeal secretions were collected and cultured on days 0, 2, and 4, and at discharge. RESULTS: The 2 groups had similar demographic characteristics, preexisting underlying diseases, and pharmacological, nutritional, and ventilatory support. Gram-negative bacteria were the predominant pathogens: Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Enter-obacter species. The 2 groups did not differ significantly in the colonization of normal (P= .72) or pathogenic (P= .62) flora, in the duration of mechanical ventilation (P= .67), or in length of stay in the intensive care (P= .22). CONCLUSION: Use of chlorhexidine combined with nonpharmacological oral care did not decrease the colonization profile, duration of mechanical ventilation, or length of stay in critically ill children receiving mechanical ventilation. PMID- 19556412 TI - Systemic inflammatory response syndrome score and race as predictors of length of stay in the intensive care unit. AB - BACKGROUND: Identifying predictors of length of stay in the intensive care unit can help critical care clinicians prioritize care in patients with acute, life threatening injuries. OBJECTIVE: To determine if systemic inflammatory response syndrome scores are predictive of length of stay in the intensive care unit in patients with acute, life-threatening injuries. METHODS: Retrospective chart reviews were completed on patients with acute, life-threatening injuries admitted to the intensive care unit at a level I trauma center in the southeastern United States. All 246 eligible charts from the trauma registry database from 1998 to 2007 were included. Systemic inflammatory response syndrome scores measured on admission were correlated with length of stay in the intensive care unit. Data on race, sex, age, smoking status, and injury severity score also were collected. Univariate and multivariate regression modeling was used to analyze data. RESULTS: Severe systemic inflammatory response syndrome scores on admission to the intensive care unit were predictive of length of stay in the unit (F=15.83; P<.001), as was white race (F=9.7; P=.002), and injury severity score (F=20.23; P<.001). CONCLUSIONS: Systemic inflammatory response syndrome scores can be measured quickly and easily at the bedside. Data support use of the score to predict length of stay in the intensive care unit. PMID- 19556414 TI - Spirituality and support for family presence during invasive procedures and resuscitations in adults. AB - BACKGROUND: Many health care professionals believe that they provide holistic care. The role of spirituality, a known variable of holism, has not been explored in relation to the support among health care professionals for family presence during invasive procedures and resuscitative efforts in adults. OBJECTIVE: To determine the relationship between spirituality of health care professionals and their support for family presence during invasive procedures and resuscitative efforts in adults. METHODS: In this descriptive correlational study, 108 participants (physicians, physician assistants, and nurses) completed the Howden Spirituality Assessment Scale and a survey to measure their support for family presence. RESULTS: A significant positive relationship was found between spirituality and support for family presence during resuscitative efforts in adults (r = 0.24, P = .05) and a significant negative correlation was found between support for family presence and the age of the health care professional (r = - 0.27, P = .01). No significant correlations were found between any of the study variables and invasive procedures in adults. CONCLUSIONS: Adopting a more holistic perspective may support family presence, especially during resuscitative efforts in adults. Allowing the option for patients' families to remain present promotes holistic family-centered care. PMID- 19556413 TI - Relationship of depressive symptoms to the impact of physical symptoms on functional status in women with heart failure. AB - BACKGROUND: Among patients with heart failure, women have worse functional status than do men, but little research has focused on determining factors that influence functional status in either sex. OBJECTIVES: To compare factors that influence functional status in men and women with heart failure and to test whether depressive symptoms mediate the relationship between physical symptoms and functional status. METHODS: A cross-sectional, descriptive study design was used. A total of 231 patients, 133 men and 98 women, were recruited from an inpatient heart failure clinic in South Korea. Functional status (the Korean Activity Scale/Index), physical symptoms (the Symptom Status Questionnaire), depressive symptoms (the Beck Depression Inventory), and situational factors (living status, socioeconomic status) were measured. Hierarchical multiple regression and mediation analysis were used for data analysis. RESULTS: Women (mean score, 24.5; SD, 17.3) had worse functional status than did men (mean score, 31.9; SD, 20.1; P = .004). Dyspnea on exertion (beta = -0.16), ankle swelling (beta = -0.19), fatigue (beta = -0.20), and depressive symptoms (beta = 0.19) were independently associated with functional status in women, whereas only dyspnea on exertion (beta = -0.30) influenced functional status of men in hierarchical multiple regression analysis. Mediation analysis indicated that depressive symptoms mediated the relationship between physical symptoms and functional status in women with heart failure, but not in men. CONCLUSIONS: Distinct physical and psychological symptoms influence functional status in women with heart failure. A systematic multidimensional intervention may be required to target depressive symptoms to improve functional status in women with heart failure. PMID- 19556415 TI - Oral health, ventilator-associated pneumonia, and intracranial pressure in intubated patients in a neuroscience intensive care unit. AB - BACKGROUND: Although oral health affects systemic health, studies of oral health during intubation among critically ill neuroscience patients are lacking. Furthermore, the effect of oral care on intracranial pressure among critically ill patients in a neuroscience intensive care unit is unknown. OBJECTIVES: To describe changes in oral health and development of ventilator-associated pneumonia during intubation among patients in a neuroscience intensive care unit and to assess the influence of oral care on intracranial pressure. METHODS: Data on 45 consecutive intubated patients admitted to a neuroscience intensive care unit during 1 year were collected by using oral cultures and the Oral Assessment Guide throughout intubation and 48 hours after extubation. Occurrence of ventilator-associated pneumonia and intracranial pressures associated with oral care were recorded. RESULTS: Oral health, assessed by the Oral Assessment Guide, deteriorated significantly during intubation and improved to almost baseline levels 48 hours after extubation. During intubation, occurrence of oral gram negative bacteria and yeast increased. The incidence of ventilator-associated pneumonia was 24% among patients enrolled for 4 to 10 days. During or after 879 instances of oral care, overall intracranial pressure did not increase. Among 30 instances in which intracranial pressure was greater than 20 mm Hg before oral care, pressure decreased during and 30 minutes after the procedure (P < .001). CONCLUSIONS: Intubation may contribute to worsening of oral health among patients in neuroscience intensive care units. Execution of oral care does not seem to affect intracranial pressure adversely. Oral care should be explored further to promote good oral and systemic health in patients in neuroscience intensive care units and to determine its effect on ventilator-associated pneumonia. PMID- 19556416 TI - Questions on organ donation and hastening death. PMID- 19556417 TI - Long pause. PMID- 19556418 TI - Acute ST-segment elevation myocardial infarction associated with diclofenac induced anaphylaxis: case report. AB - A patient in whom acute myocardial infarction developed during diclofenac-induced anaphylaxis is described. ST-segment elevation myocardial infarction is a rare complication of anaphylactic reactions, but can occur even in patients with angiographically normal coronary arteries. Physicians should be aware of such a complication in order to diagnose it early and treat it properly. In the patient described here, according to the temporal relationship with diclofenac intake and the exclusion of coronary stenosis, it is probable that diclofenac caused the symptoms. To our knowledge, this is the first reported case of Kounis syndrome due to diclofenac. The patient's recovery was uneventful. PMID- 19556420 TI - Activation of liver X receptor regulates substrate oxidation in white adipocytes. AB - Liver X receptors (LXRs) are nuclear receptors with established roles in cholesterol, lipid, and carbohydrate metabolism, although their function in adipocytes is not well characterized. Increased adipose tissue mass in obesity is associated with increased adipocyte lipolysis. Fatty acids (FA) generated by lipolysis can be oxidized by mitochondrial beta-oxidation, reesterified, or released from the adipocyte. The latter results in higher circulating levels of free FAs, in turn causing obesity-related metabolic complications. However, mitochondrial beta-oxidation can at least in part counteract an increased output of FA into circulation. In this study, we provide evidence that activation of LXRs up-regulates mitochondrial beta-oxidation in both human and murine white adipocytes. We also show that the expression of a kinase regulating the cellular fuel switch, pyruvate dehydrogenase kinase 4 (PDK4), is up-regulated by the LXR agonist GW3965 in both in vitro differentiated human primary adipocytes and differentiated murine 3T3-L1 cells. Moreover, activation of LXR causes PDK4 dependent phosphorylation of the pyruvate dehydrogenase complex, thereby decreasing its activity and attenuating glucose oxidation. The specificity of the GW3965 effect on oxidation was confirmed by RNA interference targeting LXRs. We propose that LXR has an important role in the regulation of substrate oxidation and the switch between lipids and carbohydrates as cellular fuel in both human and murine white adipocytes. PMID- 19556419 TI - Chorionic gonadotropin regulates prostaglandin E synthase via a phosphatidylinositol 3-kinase-extracellular regulatory kinase pathway in a human endometrial epithelial cell line: implications for endometrial responses for embryo implantation. AB - Successful implantation necessitates modulation of the uterine environment by the embryo for a specific period of time during the menstrual cycle. Infusion of chorionic gonadotropin (CG) into the oviducts of baboons to mimic embryo transit induces a myriad of morphological, biochemical, and molecular changes in the endometrium. Endometrial epithelial cells from both baboons and humans when stimulated by CG in vitro, activates a cAMP-independent MAPK pathway leading to prostaglandin E(2) (PGE(2)) synthesis. This study shows that in the human endometrial cell line, HES, CG, acting via its G-protein coupled receptor, phosphorylates protein kinase B, c-Raf, and ERK1/2 in a phosphatidylinositol 3 kinase (PI3K)-dependent manner. Furthermore, ERK1/2 phosphorylation is independent of the signaling paradigms of Galpha(s), Galpha(I), and epidermal growth factor receptor (EGFR) transactivation, typical of gonadal cells, indicating an alternative signaling pattern in the endometrium. After phosphorylation by CG, ERK1/2 translocates to the nucleus in a time-dependent manner. Downstream of ERK1/2, CG activates the nuclear transcription factor, Elk1, also in a PI3K-MAPK-dependent manner. Lastly, we show that in HES cells, this pathway regulates the expression of the microsomal enzyme PGE(2) synthase (mPTGES), a terminal prostanoid synthase responsible for PGE(2) synthesis. CG regulates the mPTGES promoter and also induces mPTGES synthesis in HES cells via the PI3K-ERK1/2 pathway. We suggest that this alternative PI3K-ERK-Elk pathway activated by CG regulates prostaglandin production by the endometrial epithelium and serves as an early trigger to prepare the endometrium for implantation. PMID- 19556421 TI - Inhibition of nuclear factor-kappaB or Bax prevents endoplasmic reticulum stress- but not nitric oxide-mediated apoptosis in INS-1E cells. AB - Accumulating evidence suggests that endoplasmic reticulum (ER) stress by mechanisms that include ER Ca(2+) depletion via NO-dependent down-regulation of sarcoendoplasmic reticulum Ca(2+) ATPase 2b (SERCA2b) contributes to beta-cell death in type 1 diabetes. To clarify whether the molecular pathways elicited by NO and ER Ca(2+) depletion differ, we here compare the direct effects of NO, in the form of the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP), with the effects of SERCA2 inhibitor thapsigargin (TG) on MAPK, nuclear factor kappaB (NFkappaB), Bcl-2 proteins, ER stress, and apoptosis. Exposure of INS-1E cells to TG or SNAP caused caspase-3 cleavage and apoptosis. Both TG and SNAP induced activation of the proapoptotic transcription factor CCAAT/enhancer-binding protein homologous protein (CHOP). However, other classical ER stress-induced markers such as up-regulation of ER chaperone Bip and alternative splicing of the transcription factor Xbp-1 were exclusively activated by TG. TG exposure caused NFkappaB activation, as assessed by IkappaB degradation and NFkappaB DNA binding. Inhibition of NFkappaB or the Bcl-2 family member Bax pathways protected beta cells against TG- but not SNAP-induced beta-cell death. These data suggest that NO generation and direct SERCA2 inhibition cause two quantitative and qualitative different forms of ER stress. In contrast to NO, direct ER stress induced by SERCA inhibition causes activation of ER stress signaling pathways and elicit proapoptotic signaling via NFkappaB and Bax. PMID- 19556423 TI - Repression of BMI1 in normal and leukemic human CD34(+) cells impairs self renewal and induces apoptosis. AB - High expression of BMI1 in acute myeloid leukemia (AML) cells is associated with an unfavorable prognosis. Therefore, the effects of down-modulation of BMI1 in normal and leukemic CD34(+) AML cells were studied using a lentiviral RNA interference approach. We demonstrate that down-modulation of BMI1 in cord blood CD34(+) cells impaired long-term expansion and progenitor-forming capacity, both in cytokine-driven liquid cultures as well as in bone marrow stromal cocultures. In addition, long-term culture-initiating cell frequencies were dramatically decreased upon knockdown of BMI1, indicating an impaired maintenance of stem and progenitor cells. The reduced progenitor and stem cell frequencies were associated with increased expression of p14ARF and p16INK4A and enhanced apoptosis, which coincided with increased levels of intracellular reactive oxygen species and reduced FOXO3A expression. In AML CD34(+) cells, down-modulation of BMI1 impaired long-term expansion, whereby self-renewal capacity was lost, as determined by the loss of replating capacity of the cultures. These phenotypes were also associated with increased expression levels of p14ARF and p16INK4A. Together our data indicate that BMI1 expression is required for maintenance and self-renewal of normal and leukemic stem and progenitor cells, and that expression of BMI1 protects cells against oxidative stress. PMID- 19556422 TI - Biochemical factors governing the steady-state estrone/estradiol ratios catalyzed by human 17beta-hydroxysteroid dehydrogenases types 1 and 2 in HEK-293 cells. AB - Human 17beta-hydroxysteroid dehydrogenase types 1 and 2 (17betaHSD1 and 17betaHSD2) regulate estrogen potency by catalyzing the interconversion of estrone (E1) and estradiol (E2) using nicotinamide adenine dinucleotide (phosphate) cofactors NAD(P)(H). In intact cells, 17betaHSD1 and 17betaHSD2 establish pseudo-equilibria favoring E1 reduction or E2 oxidation, respectively. The vulnerability of these equilibrium steroid distributions to mutations and to altered intracellular cofactor abundance and redox state, however, is not known. We demonstrate that the equilibrium E2/E1 ratio achieved by 17betaHSD1 in intact HEK-293 cell lines is progressively reduced from 94:6 to 10:90 after mutagenesis of R38, which interacts with the 2'-phosphate of NADP(H), and by glucose deprivation, which lowers the NADPH/NADP(+) ratio. The shift to E2 oxidation parallels changes in apparent K(m) values for purified 17betaHSD1 proteins to favor NAD(H) over NADP(H). In contrast, mutagenesis of E116 (corresponding to R38 in 17betaHSD1) and changes in intracellular cofactor ratios do not alter the greater than 90:10 E1/E2 ratio catalyzed by 17betaHSD2, and these mutations lower the apparent K(m) of recombinant 17betaHSD2 for NADP(H) only less than 3-fold. We conclude that the equilibrium E1/E2 ratio maintained by human 17betaHSD1 in intact cells is governed by NADPH saturation, which is strongly dependent on both R38 and high intracellular NADPH/NADP(+) ratios. In contrast, the preference of 17betaHSD2 for E2 oxidation strongly resists alteration by genetic and metabolic manipulations. These findings suggest that additional structural features, beyond the lack of a specific arginine residue, disfavor NADPH binding and thus support E2 oxidation by 17betaHSD2 in intact cells. PMID- 19556424 TI - Interleukin-27 inhibition of HIV-1 involves an intermediate induction of type I interferon. AB - Infection of CD4(+) chemokine coreceptor(+) targets by HIV is aided and abetted by the proficiency of HIV in eliminating or neutralizing host cell-derived defensive molecules. Among these innate protective molecules, a family of intracellular apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like (APOBEC) cytidine deaminases, is constitutively expressed but inactivated by HIV viral infectivity factor. The ability of interferon-alpha (IFN-alpha) to augment cytidine deaminases offered the possibility that the balance between virus and target cell might be altered in favor of the host. Further characterization of transcriptional profiles induced by IFN-alpha using microarrays, with the intention to identify and dissociate retroviral countermaneuvers from associated toxicities, revealed multiple molecules with suspected antiviral activity, including IL-27. To establish whether IFN-alpha toxicity might be sidestepped through the use of downstream IL-27 against HIV, we examined whether IL-27 directly regulated cytidine deaminases. Although IL-27 induces APOBECs, it does so in a delayed fashion. Dissecting the underlying regulatory events uncovered an initial IL-27-dependent induction of IFN-alpha and/or IFN-beta, which in turn, induces APOBEC3, inhibited by IFN-alpha/beta receptor blockade. In addition to macrophages, the IL-27-IFN-alpha connection is operative in CD4(+) T cells, consistent with an IFN-alpha-dependent pathway underlying host cell defense to HIV. PMID- 19556426 TI - How I treat mantle cell lymphoma. AB - Mantle cell lymphoma is included in the World Health Organization classification as distinct lymphoma subtype characterized by the t(11;14)(q13;q32) translocation, which results in overexpression of Cyclin D1. The clinical presentation often includes extranodal involvement, particularly of the bone marrow and gut. The prognosis of patients with mantle cell lymphoma (median overall survival, 3-5 years) is poorest among B-cell lymphoma patients, even though a prospectively difficult to identify subgroup can survive for years with little or no treatment. Conventional chemotherapy is not curative but obtains frequent remissions (60%-90%) which are usually shorter (1-2 years) compared with other lymphoma entities. Very intensive regimens, including autologous and allogeneic stem cell transplantation, seem required to improve the outcome, but with the median age of diagnosis being 60 years or more, such approaches are feasible only in a limited proportion of patients. The possibility of treating patients based on prognostic factors needs to be investigated prospectively. PMID- 19556427 TI - Teachers' perception of satisfaction, skill development, and skill application after instructional consultation services. AB - A primary hypothesized outcome of consultee-centered consultation, including instructional consultation (IC), is that consultees will become more skilled. However, these claims have not been well researched. Data from 274 teachers implementing IC were analyzed to investigate perceptions of satisfaction and skill development. Results indicated that teachers were highly satisfied, perceived outcomes to meet or exceed their expectations, and felt confident about handling similar problems in the future. The majority reported learning one or more skills or strategies from participating and indicated generalization of skills learned from IC to other students. Relationships between satisfaction, generalization, and perceived outcomes are also presented. Although based on descriptive methodology, this analysis of teachers' perceptions of IC provides a window into their experiences. PMID- 19556425 TI - Granulocyte-macrophage colony-stimulating factor (GM-CSF)-secreting cellular immunotherapy in combination with autologous stem cell transplantation (ASCT) as postremission therapy for acute myeloid leukemia (AML). AB - Preclinical models have demonstrated the efficacy of granulocyte-macrophage colony-stimulating factor-secreting cancer immunotherapies (GVAX platform) accompanied by immunotherapy-primed lymphocytes after autologous stem cell transplantation in hematologic malignancies. We conducted a phase 2 study of this combination in adult patients with acute myeloid leukemia. Immunotherapy consisted of autologous leukemia cells admixed with granulocyte-macrophage colony stimulating factor-secreting K562 cells. "Primed" lymphocytes were collected after a single pretransplantation dose of immunotherapy and reinfused with the stem cell graft. Fifty-four subjects were enrolled; 46 (85%) achieved a complete remission, and 28 (52%) received the pretransplantation immunotherapy. For all patients who achieved complete remission, the 3-year relapse-free survival (RFS) rate was 47.4% and overall survival was 57.4%. For the 28 immunotherapy-treated patients, the RFS and overall survival rates were 61.8% and 73.4%, respectively. Posttreatment induction of delayed-type hypersensitivity reactions to autologous leukemia cells was associated with longer 3-year RFS rate (100% vs 48%). Minimal residual disease was monitored by quantitative analysis of Wilms tumor-1 (WT1), a leukemia-associated gene. A decrease in WT1 transcripts in blood was noted in 69% of patients after the first immunotherapy dose and was also associated with longer 3-year RFS (61% vs 0%). In conclusion, immunotherapy in combination with primed lymphocytes and autologous stem cell transplantation shows encouraging signals of potential activity in acute myeloid leukemia (ClinicalTrials.gov: NCT00116467). PMID- 19556428 TI - A literacy tutoring experience for prospective special educators and struggling second graders. AB - This study examined the learning of teacher candidates taking a language arts course in a special-educator preparation program and that of the second graders they tutored in a supervised field component of the course. Teacher candidates' knowledge of literacy instruction was assessed using five knowledge tasks; children were assessed on several measures of basic reading and spelling skills as well as on their knowledge of phonics concepts such as syllable types. Teacher candidates generally had inaccurate perceptions of their knowledge at pretest, but their knowledge improved significantly on all tasks after course instruction. Tutored children improved significantly from pre- to posttest on all assessments. The study suggests that carefully designed literacy coursework with field experiences can benefit both prospective special educators and struggling readers. PMID- 19556429 TI - CEBPA resembles Roman god Janus. PMID- 19556430 TI - Cancer & inherited bone marrow failure states. PMID- 19556431 TI - Functional neutrophils from human ES cells. PMID- 19556432 TI - UCB transplantation: miRNA involvement. PMID- 19556433 TI - Micro-classifying diffuse large B-cell lymphomas. PMID- 19556434 TI - Muscle iron in stress erythropoiesis? PMID- 19556435 TI - Myeloid cell-induced angiogenesis: a sticky business. PMID- 19556436 TI - Cryopreserved ovarian tissues can maintain a long-term function after heterotopic autotransplantation in rat. AB - The functional longevity of cryopreserved ovarian grafts is one of the most challenging questions regarding ovarian transplantation at present. This study used a rat ovarian grafting model to investigate whether ovarian tissues from adult rats, which had been cryopreserved by vitrification and followed by heterotopic transplantation, could establish long-term hormone secretion and follicle development. Fresh and cryopreserved ovarian tissues were autologously transplanted under the kidney capsule. One-third of the animals in each group (sham-operated, fresh autografts, cryopreserved autografts, or castrated) were killed 5, 8, or 10 months after transplantation. Vaginal cytology, serum estradiol (E(2)), progesterone, and the morphology of the reproductive tract were used to assess ovarian function. Both fresh and cryopreserved ovarian grafts survived well in all the animal models with comparable proportion of follicles at each stage of folliculogenesis at all three time points. The serum E(2) and progesterone concentrations in the groups with fresh or cryopreserved grafts remained comparable with those in sham-operated controls at all investigated time points. However, a loss of grafts and primordial follicles following heterotopic transplantation was noted. In conclusion, the heterotopic autotransplantation of vitrified ovarian tissues from adult rat without vascular anastomosis can maintain long-term ovarian function and exert endocrine function in target organs, in spite of the reduction in follicle pool. PMID- 19556437 TI - Live offspring from vitrified blastocysts derived from fresh and cryopreserved ovarian tissue grafts of adult mice. AB - Ovarian tissue cryopreservation and transplantation can be used to preserve fertility for cancer patients. In this study, we assessed the viability and function of ovarian tissue from adult mice that was cryopreserved by solid surface vitrification or traditional slow-cooling using various in vitro and in vivo techniques, including allotransplantation, in vitro oocyte maturation, embryo culture in vitro, blastocyst cryopreservation, embryo transfer, and development. The importance of cumulus cells for oocyte maturation, fertilization, and embryo development was investigated. Graft recovery, follicle survival, and oocyte retrieval was similar in control, vitrified, and slow-cooled groups. High rates of oocyte maturation, cleavage, and blastocyst formation were achieved, with no significant differences between the control, vitrified or slow cooled ovarian tissue grafts. The presence of cumulus cells was important for oocyte maturation, fertilization, and subsequent development. Cumulus-oocyte complexes with no surrounding cumulus cells (N-COCs) or with an incomplete layer (P-COCs) had significantly lower rates of oocyte maturation and blastocyst formation than cumulus-oocyte complexes with at least one complete layer of cumulus cells (F-COCs; maturation rate: 63, 78 vs 94%; blastocyst rate: 29, 49 vs 80%). Live births were achieved using vitrified blastocysts derived from oocytes taken from vitrified and slow-cooled ovarian tissue heterotypic allografts. Successful production of healthy offspring from these vitrified blastocysts suggests that this technique should be considered as a useful stage to pause in the assisted reproduction pathway. This provides an alternative protocol for restoring fertility and offering cancer patients a better indication of their chances of pregnancy and live birth. PMID- 19556439 TI - Progesterone and conceptus elongation in cattle: a direct effect on the embryo or an indirect effect via the endometrium? AB - The steroid hormone progesterone (P(4)) plays a key role in the reproductive events associated with pregnancy establishment and maintenance. High concentrations of circulating P(4) in the immediate post-conception period have been associated with an advancement of conceptus elongation, an associated increase in interferon-tau production and higher pregnancy rates in cattle. Using in vitro and in vivo models and approximately 8500 bovine oocytes across six experiments, the aim of this study was to establish the route through which P(4) affects bovine embryo development in vitro and in vivo. mRNA for P(4) receptors was present at all stages of embryo development raising the possibility of a direct effect of P(4) on the embryo. Exposure to P(4) in vitro in the absence or presence of oviduct epithelial cells did not affect the proportion of embryos developing to the blastocyst stage, blastocyst cell number or the relative abundance of selected transcripts in the blastocyst. Furthermore, exposure to P(4) in vitro did not affect post-hatching elongation of the embryo following transfer to synchronized recipients and recovery on Day 14. By contrast, transfer of in vitro derived blastocysts to a uterine environment previously primed by elevated P(4) resulted in a fourfold increase in conceptus length on Day 14. These data provide clear evidence to support the hypothesis that P(4)-induced changes in the uterine environment are responsible for the advancement in conceptus elongation reported previously in cattle and that, interestingly, the embryo does not need to be present during the period of high P(4) in order to exhibit advanced elongation. PMID- 19556440 TI - Tight glucose control in critically ill patients: should glucose meters be used? PMID- 19556438 TI - Adenine nucleotide translocase 4 deficiency leads to early meiotic arrest of murine male germ cells. AB - Male fertility relies on the highly specialized process of spermatogenesis to continually renew the supply of spermatozoa necessary for reproduction. Central to this unique process is meiosis that is responsible for the production of haploid spermatozoa as well as for generating genetic diversity. During meiosis I, there is a dramatic increase in the number of mitochondria present within the developing spermatocytes, suggesting an increased necessity for ATP production and utilization. Essential for the utilization of ATP is the translocation of ADP and ATP across the inner mitochondrial membrane, which is mediated by the adenine nucleotide translocases (Ant). We recently identified and characterized a novel testis specific Ant, ANT4 (also known as SLC25A31 and Aac4). The generation of Ant4-deficient animals resulted in the severe disruption of the seminiferous epithelium with an apparent spermatocytic arrest of the germ cell population. In the present study utilizing a chromosomal spread technique, we determined that Ant4-deficiency results in an accumulation of leptotene spermatocytes, a decrease in pachytene spermatocytes, and an absence of diplotene spermatocytes, indicating early meiotic arrest. Furthermore, the chromosomes of Ant4-deficient pachytene spermatocyte occasionally demonstrated sustained gammaH2AX association as well as synaptonemal complex protein 1 (SYCP1)/SYCP3 dissociation beyond the sex body. Large ATP supplies from mitochondria may be critical for normal progression of spermatogenesis during early stages of meiotic prophase I, including DNA double strand break repair and chromosomal synapsis. PMID- 19556441 TI - 25-OH vitamin D3 concentrations in Chinese, Malays, and Indians. PMID- 19556442 TI - Total plasma homocysteine: the mediator/marker controversy continues. 1994. PMID- 19556443 TI - Highly sensitive cardiac troponin I assay leads to lowered specificity. PMID- 19556444 TI - New testing approach in HLA genotyping helps overcome barriers in effective clinical practice. AB - BACKGROUND: Severe and potentially fatal hypersensitivity reactions to drugs, particularly antiepileptics, are clinically unpredictable. Recent evidence has revealed a strong and specific association between the implicated drug, the type of adverse reaction, and the particular HLA genotype. An urgent need exists for rapid diagnosis of HLA status to guide drug prescription; however, traditional HLA genotyping has a long turnaround time, is expensive, and is available only in specialized centers. We tested the feasibility of the loop-mediated isothermal amplification (LAMP)-based approach to detect a specific HLA genotype. As an example, we used B*1502, an HLA allele strongly associated with carbamazepine induced Stevens-Johnson syndrome/toxic epidermal necrolysis, and validated the assay's application as a simple, accurate, rapid, and low-cost blood test for use in both clinical and bedside settings. METHODS: We evaluated B*1502 status with the new LAMP method and compared the results with those obtained by sequence based typing (SBT) (n = 250) and by sequence-specific primer PCR (SSP-PCR) (n = 200) for 450 samples of DNA (n = 50) and blood (n = 400) from a hematology laboratory. RESULTS: LAMP results showed 100% concordance with both SBT and SSP PCR results, confirming that LAMP detection of a specific HLA genotype (B*1502 in this case) is an accurate method. All results were available within 1 h. CONCLUSIONS: Our study confirmed that the new LAMP method for detecting a specific HLA genotype is simple, inexpensive, accurate, and rapid, and may be of help in overcoming barriers in effective clinical practice. PMID- 19556445 TI - The international antidoping system and why it works. PMID- 19556446 TI - Myeloperoxidase: a useful biomarker for cardiovascular disease risk stratification? AB - BACKGROUND: Inflammation and oxidative stress are associated with atherosclerosis. Myeloperoxidase (MPO) is linked to both inflammation and oxidative stress by its location in leukocytes and its role in catalyzing the formation of oxidizing agents. Recent evidence suggests that MPO activity precipitates atherogenesis. Measurement of MPO in plasma may therefore contribute to cardiovascular disease (CVD) risk stratification. CONTENT: Cross-sectional studies, case-control studies, and prospective-cohort studies investigating the relation between MPO and CVD have been evaluated. Differences in study populations, sample materials, sample handling, and assays were ascertained. Potential causal mechanisms linking MPO to accelerated atherosclerosis are discussed here. A majority of studies indicate that measurement of MPO in plasma was associated with improved CVD risk stratification above and beyond risk stratification results obtained with markers used in routine clinical practice. However, comparison of these epidemiological studies with regard to MPO and outcome is hampered because the reported MPO concentration depends on the assay method, sampling material, and preanalytical and analytical procedures. The link between MPO and CVD can, at least partly, be explained by MPO-dependent oxidation of LDL and HDL, subsequently leading to cholesterol accumulation in the arterial wall. Furthermore, MPO may reduce the bioavailability of nitric oxide, resulting in endothelial dysfunction. Finally, MPO destabilizes atherosclerotic plaques. SUMMARY: Increasing evidence suggests that MPO is causally linked to atherosclerosis and its measurement may improve CVD risk estimation. Before MPO can be used in routine clinical practice, however, standardization of sampling and laboratory procedures is needed. PMID- 19556448 TI - Use of saliva-based nano-biochip tests for acute myocardial infarction at the point of care: a feasibility study. AB - BACKGROUND: For adults with chest pain, the electrocardiogram (ECG) and measures of serum biomarkers are used to screen and diagnose myocardial necrosis. These measurements require time that can delay therapy and affect prognosis. Our objective was to investigate the feasibility and utility of saliva as an alternative diagnostic fluid for identifying biomarkers of acute myocardial infarction (AMI). METHODS: We used Luminex and lab-on-a-chip methods to assay 21 proteins in serum and unstimulated whole saliva procured from 41 AMI patients within 48 h of chest pain onset and from 43 apparently healthy controls. Data were analyzed by use of logistic regression and area under curve (AUC) for ROC analysis to evaluate the diagnostic utility of each biomarker, or combinations of biomarkers, in screening for AMI. RESULTS: Both established and novel cardiac biomarkers demonstrated significant differences in concentrations between patients with AMI and controls without AMI. The saliva-based biomarker panel of C reactive protein, myoglobin, and myeloperoxidase exhibited significant diagnostic capability (AUC = 0.85, P < 0.0001) and in conjunction with ECG yielded strong screening capacity for AMI (AUC = 0.96) comparable to that of the panel (brain natriuretic peptide, troponin-I, creatine kinase-MB, myoglobin; AUC = 0.98) and far exceeded the screening capacity of ECG alone (AUC approximately 0.6). En route to translating these findings to clinical practice, we adapted these unstimulated whole saliva tests to a novel lab-on-a-chip platform for proof-of principle screens for AMI. CONCLUSIONS: Complementary to ECG, saliva-based tests within lab-on-a-chip systems may provide a convenient and rapid screening method for cardiac events in prehospital stages for AMI patients. PMID- 19556449 TI - Molecular cloning and pharmacological characterization of monkey MT1 and MT2 melatonin receptors showing high affinity for the agonist ramelteon. AB - Melatonin receptor agonists such as melatonin and ramelteon [(S)-N-[2-(1,6,7,8 tetrahydro-2H-indeno-[5,4-b]furan-8-yl)ethyl]-propionamide; TAK-375] have sleep promoting effects in humans. In preclinical models, these effects are more similar to those observed in monkeys than in other species. However, in contrast to the human melatonin receptors, the pharmacological characteristics of the monkey melatonin receptors have yet to be elucidated. In this study, we cloned the cynomolgus monkey MT(1) and MT(2) melatonin receptors based on rhesus monkey genome sequences and then characterized the monkey melatonin receptors and compared their pharmacological properties with those of the human homologs. The overall amino acid sequences of the monkey MT(1) and MT(2) melatonin receptors showed high homology to the human MT(1) (95%) and MT(2) (96%) receptors, respectively. Saturation binding experiments with 2-[(125)I]iodomelatonin revealed that the dissociation constants (K(d)) for the monkey MT(1) and MT(2) melatonin receptors were 19.9 and 70.4 pM, respectively. In ligand competition assays using 2-[(125)I]iodomelatonin, ramelteon displayed approximately 3- to 7 fold higher affinities than melatonin for the recombinant monkey MT(1) and MT(2) melatonin receptors and monkey suprachiasmatic nucleus membranes. This higher affinity of ramelteon compared with melatonin has also been observed in human melatonin receptors. Furthermore, ramelteon inhibited pituitary adenylate cyclase activating polypeptide-27-stimulated cAMP production with higher potency than melatonin. In conclusion, this information will help us to understand the pharmacological effects of melatonin receptor agonists in monkeys. PMID- 19556450 TI - Differential effects of p38 mitogen-activated protein kinase and cyclooxygenase 2 inhibitors in a model of cardiovascular disease. AB - The evidence is compelling for a role of inflammation in cardiovascular diseases; however, the chronic use of anti-inflammatory drugs for these indications has been disappointing. The recent study compares the effects of two anti inflammatory agents [cyclooxygenase 2 (COX2) and p38 inhibitors] in a model of cardiovascular disease. The vascular, renal, and cardiac effects of 4-(4 methylsulfonylphenyl)-3-phenyl-5H-furan-2-one (rofecoxib; a COX2 inhibitor) and 6 {5-[(cyclopropylamino)carbonyl]-3-fluoro-2-methylphenyl}-N-(2,2-dimethylpropyl)-3 pyridinecarboxamide [GSK-AHAB, a selective p38 mitogen-activated protein kinase (MAPK) inhibitor], were examined in the spontaneously hypertensive stroke-prone rat (SHR-SP). In SHR-SPs receiving a salt-fat diet (SFD), chronic treatment with GSK-AHAB significantly and dose-dependently improved survival, endothelial dependent and -independent vascular relaxation, and indices of renal function, and it attenuated dyslipidemia, hypertension, cardiac remodeling, plasma renin activity (PRA), aldosterone, and interleukin-1beta (IL-1beta). In contrast, chronic treatment with a COX2-selective dose of rofecoxib exaggerated the harmful effects of the SFD, i.e., increasing vascular and renal dysfunction, dyslipidemia, hypertension, cardiac hypertrophy, PRA, aldosterone, and IL-1beta. The protective effects of a p38 MAPK inhibitor are clearly distinct from the deleterious effects of a selective COX2 inhibitor in the SHR-SP and suggest that anti-inflammatory agents can have differential effects in cardiovascular disease. The results also suggest a method for evaluating long-term cardiovascular efficacy and safety. PMID- 19556452 TI - Heat stress increases myonuclear number and fiber size via satellite cell activation in rat regenerating soleus fibers. AB - To investigate the effects of heat stress (hyperthermia) on muscle degeneration regeneration, the soleus muscles of adult male Wistar rats were injected bilaterally with a single injection of bupivacaine. The rats were assigned to a sedentary control (Con), heat stress (Heat), bupivacaine-injected (BPVC), or bupivacaine-injected plus heat stress (BPVC+Heat) group. Heat stress was induced in the Heat and BPVC+Heat groups by immersion of the lower half of the body into water maintained at 42 +/- 1 degrees C for 30 min 48 h after the injection of bupivacaine and every other day during the following 1 or 2 wk. The soleus muscles in all groups were excised 24 h after the final bout of heat stress. Mean muscle weight, fiber cross-sectional area, myonuclear number, and heat shock protein 72 (Hsp72) and calcineurin protein levels were lower in the BPVC than in the Con or Heat groups at both time points. In contrast, several of these parameters in the BPVC+Heat group were not different or higher than in the Con or Heat groups at the 1- and/or 2-wk time points. The number of total and activated satellite cells, estimated by analyses of Pax7-negative, M-cadherin-negative, and MyoD-positive nuclei, was greater in BPVC+Heat than in all other groups. Combined, the results indicate that heat stress-related activation of satellite cells and upregulation of Hsp72 and calcineurin expression played important roles in the regeneration of the soleus fibers after bupivacaine injection. PMID- 19556451 TI - Alpha2-adrenergic receptors attenuate secretagogue-induced endocytosis and promote exocytosis of intestinal NHE2 and NHE3. AB - Adrenergic agonists, through activation of intestinal epithelial alpha2 adrenergic receptors (alpha2AR), inhibit electrolyte secretion and promote absorption. The mechanisms of action to promote basal Na(+) absorption and inhibit stimulated secretion are not understood completely. The effects of alpha2 agonists on Na(+) transport were studied in a cell line, Caco2-3B, derived from the Caco2 cell line engineered to permanently express human alpha2A-adrenergic receptors. Serosal, but not mucosal, addition of the alpha2AR agonist N-(2,6 dichlorophenyl)-4,5-dihydro-1H-imidazol-2-amine (clonidine) increased Caco2-3B apical (22)Na(+) uptake, an effect not seen in the Caco2 parent line that lacks alpha2AR expression. This effect was blocked by the alpha2AR antagonist 17alpha yohmban-16alpha-carboxylic acid methyl ester (yohimbine). Increased Na(+) uptake was paralleled by increased apical surface abundance of the sodium/hydrogen exchangers NHE2 and NHE3. No changes in total cell NHE2 and NHE3 expression were observed. Clonidine also inhibited both cAMP and Ca(2+)-induced decreases in apical Na(+) uptake and apical membrane NHE2 and NHE3 endocytosis stimulated by these agents. alpha2AR actions were mediated via stimulation of phospholipase C, and metabolism of arachidonic acid by an epoxygenase activity followed epidermal growth factor release and activation of the epidermal growth factor receptor, resulting in phosphatidylinositol-3-kinase and Akt stimulation. In summary, activation of intestinal epithelial alpha2AR significantly blocks the inhibition of apical Na(+) transporters by cAMP- and Ca(2+)-mediated pathways and also directly increases apical sodium/hydrogen exchange activities. By both blocking electrolyte secretion and promoting absorption, alpha2-agonists could be potent antidiarrheal agents that could directly counteract the actions of toxigenic pathogens and other secretagogues causing secretory diarrhea. PMID- 19556453 TI - Exercise training decreases the size and alters the composition of the neointima in a porcine model of percutaneous transluminal coronary angioplasty (PTCA). AB - Exercise training (EX) following percutaneous transluminal coronary angiography (PTCA) reduces progression to restenosis and increases event-free survival rates. Our aim was to determine whether EX inhibits lesion development and/or alters the extracellular matrix (ECM) composition of the neointima (NI) in a porcine PTCA model. Miniature Yucatan swine were assigned to cage confinement (SED) or EX for 20 wk. After 16 wk, all animals underwent a PTCA procedure of the left anterior descending artery (LAD) and left circumflex artery (LCX), with subsequent placement of an externalized jugular catheter. Animals recovered for 2 days and then resumed the previous protocol of SED or EX. Twelve days following PTCA, all animals received an intravenous bromodeoxyuridine (BrdU) injection to label proliferating cells. At 28 days following PTCA, the animals were euthanized, the LAD and LCX excised, and underwent standard histological processing for total collagen, type I collagen, fibronectin, BrdU, and Verhoeff-van Gieson stain. Our results demonstrate that EX significantly decreased lesion size and NI proliferation (-48%) in the LAD (P < 0.05) but not the LCX. Furthermore, EX attenuated type I collagen expression only in LAD, whereas total collagen was increased (5.9%) and fibronectin was decreased (-7.9%) in the NI of both vessels (P < 0.05). In conclusion, EX following PTCA may increase event-free survival rates following PTCA by decreasing lesion size and altering ECM composition. PMID- 19556454 TI - Sex hormone effects on body fluid and sodium regulation in women with and without exercise-associated hyponatremia. AB - We hypothesized that exercise-associated hyponatremia (EAH) is a function of excess sodium loss combined with high water intake in women at risk for dysnatremias during endurance exercise. We further hypothesized that estradiol and progesterone exposure increases fluid retention and sodium loss during exercise in women at risk for EAH. For 16 days we suppressed estrogens and progesterone with a gonadotropin-releasing hormone antagonist (GnRH ant) in seven women with (Hypo) and nine women without (no Hypo) a history of hyponatremia; we added 17beta-estradiol (0.2 mg/day patches) for days 4-16 (E(2)) and progesterone (200 mg/day) for days 13-16 (E(2)-P(4)). Under each hormone condition, subjects cycled in 35 degrees C at 65% peak oxygen consumption (Vo(2peak)) for 60 min, then at 55-60% Vo(2peak) for 120 min. Subjects drank 8 ml/kg of water (and replenished urine volume) every 30 min over the final 120 min of exercise. S([Na+]) fell by 4.3, 3.9, and 3.1 meq/l (P < 0.05) with drinking during GnRH ant, E(2), and E(2)-P(4) in Hypo, with little fall in no Hypo. Under all conditions, combined urine and sweat sodium loss were similar between Hypo [-85.6 (SD 36.2), -86.4 (SD 39.2), and -112.0 (SD 30.0) meq] and no Hypo [-98.0 (SD 54.8), -80.9 (SD 57.6), and -105.1 (SD 46.4) meq, for GnRH, E2, and E2-P4], as was mass balance of electrolytes (E(MB)) for Hypo [-104.8 (SD 32.8), -103.6 (SD 42.1), and -132.8 (SD 34.9) meq] compared with no Hypo [-128.8 (SD 57.2), -113.5 (SD 61.1), and -143.4 (SD 49.6) meq for GnRH, E2, and E2-P4]. Mass balance of water [V(MB), for Hypo, 0.42 (SD 0.10), 0.62 (SD 0.25), and -0.11 (SD 0.11) liter] compared with no Hypo [0.01 (SD 0.15), 0.03 (SD 17), and -0.16 (SD 0.13) liter for GnRH, E2, and E2-P4, P < 0.05] indicates water retention was the primary contributor to the lower S([Na+]) in Hypo women. PMID- 19556456 TI - A spring in your step: some is good, more is not always better. PMID- 19556455 TI - Deep pulmonary lymphatics in immature lungs. AB - Recently, we found that the translocation of inhaled nanoparticles from the air space to secondary organs is age dependent and substantially greater in neonates than in adults (J Respir Crit Care Med 177: A48, 2008). One reason for this difference might be age-dependent differences in alveolar barrier integrity. Because the neonate lung is undergoing morphogenetic and fluid balance changes, we hypothesize that the alveolar barrier of developing lungs is more easily compromised and susceptible to foreign material influx than that of adult lungs. On the basis of these hypotheses, we predict that the postnatally developing lung is also more likely to allow the translocation of some materials from the air space to the lymphatic lumens. To test this idea, we intratracheally instilled methyl methacrylate into immature and adult lungs and compared lymphatic filling between these two age groups. Scanning electron microscopy of the resultant corrosion casts revealed peribronchial saccular and conduit lymphatic architecture. Deep pulmonary lymphatic casts were present on the majority (58.5%) of airways in immature lungs, but lymphatic casting in adult lungs, as anticipated, was much more infrequent (21.6%). Thus the neonate lung appears to be more susceptible than the adult lung to the passage of instilled methyl methacrylate from the air space into the lymphatics. We speculate that this could imply greater probability of translocation of other materials, such as nanoparticles, from the immature lung as well. PMID- 19556457 TI - Diurnal physiology: core principles with application to the pathogenesis, diagnosis, prevention, and treatment of myocardial hypertrophy and failure. AB - The circadian system has been shown to be fundamentally important in human health and disease. Recently, there have been major advances in our understanding of daily rhythmicity, and its relevance to human physiology, and to the pathogenesis and treatment of cardiac hypertrophy and heart failure. Cardiovascular tissues, such as heart and blood vessels, show remarkable daily variation in gene expression, metabolism, growth, and remodeling. Moreover, synchrony of daily molecular and physiological rhythms is integral to healthy organ growth and renewal. Disruption of these rhythms adversely affects normal growth, also the remodeling mechanisms in disease, leading to gross abnormalities in heart and vessels. These observations provide new insights into the pathogenesis, diagnosis, treatment, and prevention of heart disease. In this review, we focus on the recent advances in circadian biology and cardiovascular function, with particular emphasis on how this applies to human myocardial hypertrophy and heart failure, and the implications and importance for translational medicine. PMID- 19556458 TI - Mechanical properties and collagen cross-linking of the patellar tendon in old and young men. AB - Age-related loss in muscle mass and strength impairs daily life function in the elderly. However, it remains unknown whether tendon properties also deteriorate with age. Cross-linking of collagen molecules provides structural integrity to the tendon fibrils and has been shown to change with age in animals but has never been examined in humans in vivo. In this study, we examined the mechanical properties and pyridinoline and pentosidine cross-link and collagen concentrations of the patellar tendon in vivo in old (OM) and young men (YM). Seven OM (67 +/- 3 years, 86 +/- 10 kg) and 10 YM (27 +/- 2 years, 81 +/- 8 kg) with a similar physical activity level (OM 5 +/- 6 h/wk, YM 5 +/- 2 h/wk) were examined. MRI was used to assess whole tendon dimensions. Tendon mechanical properties were assessed with the use of simultaneous force and ultrasonographic measurements during ramped isometric contractions. Percutaneous tendon biopsies were taken and analyzed for hydroxylysyl pyridinoline (HP), lysyl pyridinoline (LP), pentosidine, and collagen concentrations. We found no significant differences in the dimensions or mechanical properties of the tendon between OM and YM. Collagen concentrations were lower in OM than in YM (0.49 +/- 0.27 vs. 0.73 +/- 0.14 mg/mg dry wt; P < 0.05). HP concentrations were higher in OM than in YM (898 +/- 172 vs. 645 +/- 183 mmol/mol; P < 0.05). LP concentrations were higher in OM than in YM (49 +/- 38 vs. 16 +/- 8 mmol/mol; P < 0.01), and pentosidine concentrations were higher in OM than in YM (73 +/- 13 vs. 11 +/- 2 mmol/mol; P < 0.01). These cross-sectional data raise the possibility that age may not appreciably influence the dimensions or mechanical properties of the human patellar tendon in vivo. Collagen concentration was reduced, whereas both enzymatic and nonenzymatic cross-linking of concentration was elevated in OM vs. in YM, which may be a mechanism to maintain the mechanical properties of tendon with aging. PMID- 19556459 TI - In vivo oxidative capacity varies with muscle and training status in young adults. AB - It is well established that exercise training results in increased muscle oxidative capacity. Less is known about how oxidative capacities in distinct muscles, in the same individual, are affected by different levels of physical activity. We hypothesized that 1) trained individuals would have higher oxidative capacity than untrained individuals in both tibialis anterior (TA) and vastus lateralis (VL) and 2) oxidative capacity would be higher in TA than VL in untrained, but not in trained, individuals. Phosphorus magnetic resonance spectroscopy was used to measure the rate of phosphocreatine recovery (k(PCr)), which reflects the rate of oxidative phosphorylation, following a maximal voluntary isometric contraction of the TA and VL in healthy untrained (7 women, 7 men, 25.7 +/- 3.6 yr; mean +/- SD) and trained (5 women, 7 men, 27.5 +/- 3.4 yr) adults. Daily physical activity levels were measured using accelerometry. The trained group spent threefold more time ( approximately 90 vs. approximately 30 min/day; P < 0.001) in moderate to vigorous physical activity (MVPA). Overall, k(PCr) was higher in VL than in TA (P = 0.01) and higher in trained than in untrained participants (P < 0.001). The relationship between k(PCr) and MVPA was more robust in VL (r = 0.64, P = 0.001, n = 25) than in TA (r = 0.38, P = 0.06, n = 25). These results indicate greater oxidative capacity in vivo in trained compared with untrained individuals in two distinct muscles of the lower limb and provide novel evidence of higher oxidative capacity in VL compared with TA in young humans, irrespective of training status. The basis for this difference is not known at this time but likely reflects a difference in usage patterns between the muscles. PMID- 19556460 TI - Improving assessment of daily energy expenditure by identifying types of physical activity with a single accelerometer. AB - Accelerometers are often used to quantify the acceleration of the body in arbitrary units (counts) to measure physical activity (PA) and to estimate energy expenditure. The present study investigated whether the identification of types of PA with one accelerometer could improve the estimation of energy expenditure compared with activity counts. Total energy expenditure (TEE) of 15 subjects was measured with the use of double-labeled water. The physical activity level (PAL) was derived by dividing TEE by sleeping metabolic rate. Simultaneously, PA was measured with one accelerometer. Accelerometer output was processed to calculate activity counts per day (AC(D)) and to determine the daily duration of six types of common activities identified with a classification tree model. A daily metabolic value (MET(D)) was calculated as mean of the MET compendium value of each activity type weighed by the daily duration. TEE was predicted by AC(D) and body weight and by AC(D) and fat-free mass, with a standard error of estimate (SEE) of 1.47 MJ/day, and 1.2 MJ/day, respectively. The replacement in these models of AC(D) with MET(D) increased the explained variation in TEE by 9%, decreasing SEE by 0.14 MJ/day and 0.18 MJ/day, respectively. The correlation between PAL and MET(D) (R(2) = 51%) was higher than that between PAL and AC(D) (R(2) = 46%). We conclude that identification of activity types combined with MET intensity values improves the assessment of energy expenditure compared with activity counts. Future studies could develop models to objectively assess activity type and intensity to further increase accuracy of the energy expenditure estimation. PMID- 19556461 TI - An ER-mitochondria tethering complex revealed by a synthetic biology screen. AB - Communication between organelles is an important feature of all eukaryotic cells. To uncover components involved in mitochondria/endoplasmic reticulum (ER) junctions, we screened for mutants that could be complemented by a synthetic protein designed to artificially tether the two organelles. We identified the Mmm1/Mdm10/Mdm12/Mdm34 complex as a molecular tether between ER and mitochondria. The tethering complex was composed of proteins resident of both ER and mitochondria. With the use of genome-wide mapping of genetic interactions, we showed that the components of the tethering complex were functionally connected to phospholipid biosynthesis and calcium-signaling genes. In mutant cells, phospholipid biosynthesis was impaired. The tethering complex localized to discrete foci, suggesting that discrete sites of close apposition between ER and mitochondria facilitate interorganelle calcium and phospholipid exchange. PMID- 19556462 TI - String theory, quantum phase transitions, and the emergent Fermi liquid. AB - A central problem in quantum condensed matter physics is the critical theory governing the zero-temperature quantum phase transition between strongly renormalized Fermi liquids as found in heavy fermion intermetallics and possibly in high-critical temperature superconductors. We found that the mathematics of string theory is capable of describing such fermionic quantum critical states. Using the anti-de Sitter/conformal field theory correspondence to relate fermionic quantum critical fields to a gravitational problem, we computed the spectral functions of fermions in the field theory. By increasing the fermion density away from the relativistic quantum critical point, a state emerges with all the features of the Fermi liquid. PMID- 19556463 TI - A gene network regulating lysosomal biogenesis and function. AB - Lysosomes are organelles central to degradation and recycling processes in animal cells. Whether lysosomal activity is coordinated to respond to cellular needs remains unclear. We found that most lysosomal genes exhibit coordinated transcriptional behavior and are regulated by the transcription factor EB (TFEB). Under aberrant lysosomal storage conditions, TFEB translocated from the cytoplasm to the nucleus, resulting in the activation of its target genes. TFEB overexpression in cultured cells induced lysosomal biogenesis and increased the degradation of complex molecules, such as glycosaminoglycans and the pathogenic protein that causes Huntington's disease. Thus, a genetic program controls lysosomal biogenesis and function, providing a potential therapeutic target to enhance cellular clearing in lysosomal storage disorders and neurodegenerative diseases. PMID- 19556464 TI - DICER1 mutations in familial pleuropulmonary blastoma. AB - Pleuropulmonary blastoma (PPB) is a rare pediatric lung tumor that is often part of an inherited cancer syndrome. PPBs consist of mesenchymal cells that are susceptible to malignant transformation and cysts lined by epithelial cells. In a subset of patients, overgrowth of the cysts by mesenchymal cells leads to sarcoma formation. Here, we show that 11 multiplex PPB families harbor heterozygous germline mutations in DICER1, a gene encoding an endoribonuclease critical to the generation of small noncoding regulatory RNAs. Expression of DICER1 protein was undetectable in the epithelial component of PPB tumors but was retained in the malignant mesenchyme (sarcoma). We hypothesize that loss of DICER1 in the epithelium of the developing lung alters the regulation of diffusible factors that promote mesenchymal proliferation. PMID- 19556465 TI - Measuring the cosmic-ray acceleration efficiency of a supernova remnant. AB - Cosmic rays are the most energetic particles arriving at Earth. Although most of them are thought to be accelerated by supernova remnants, the details of the acceleration process and its efficiency are not well determined. Here we show that the pressure induced by cosmic rays exceeds the thermal pressure behind the northeast shock of the supernova remnant RCW 86, where the x-ray emission is dominated by synchrotron radiation from ultrarelativistic electrons. We determined the cosmic-ray content from the thermal Doppler broadening measured with optical spectroscopy, combined with a proper-motion study in x-rays. The measured postshock proton temperature, in combination with the shock velocity, does not agree with standard shock heating, implying that >50% of the postshock pressure is produced by cosmic rays. PMID- 19556466 TI - Osteotomies about the knee for tibiofemoral malalignment in the athletic patient. AB - Osteotomies have a role in the active patient with degenerative joint disease of the medial or lateral knee who, for reasons of age or activity level, is not yet a good candidate for prosthetic arthroplasty. Recognition and treatment of malalignment associated with ligamentous instability is essential if long-term good outcomes are to be expected from ligamentous reconstruction. Also, treatment of concomitant malalignment and the unloading of the operative site is now recognized as an important adjunct to any cartilage-preserving surgery. This review examines the use of osteotomies about the knee in the athletic patient. Indications, contraindications, preoperative planning, surgical techniques, and complications are reviewed. PMID- 19556467 TI - Common carotid artery laceration in a professional hockey player: a case report. PMID- 19556468 TI - A biomechanical comparison of transosseous-suture anchor and suture bridge rotator cuff repairs in cadavers. AB - BACKGROUND: Several biomechanical studies comparing open and arthroscopic rotator cuff repair methods have shown inferior performance of arthroscopic repairs. Suture anchor-augmented transosseous repairs and suture bridge repairs have shown superior biomechanical performance when compared with other methods, but these 2 repair methods have not been directly compared. HYPOTHESIS: There will be no difference in the biomechanical performance of the transosseous-suture anchor and suture bridge techniques. STUDY DESIGN: Controlled laboratory study. METHODS: Eight paired cadaveric shoulder specimens (16 specimens) had creation followed by repair of a complete tear of the supraspinatus, with the first member of each pair undergoing repair by a transosseous-suture anchor technique and the second member undergoing repair by the suture bridge technique. Specimens were then cycled from 10 to 180 N for 200 cycles, followed by testing to failure at 33 mm/s. Elongation was measured during cyclic testing, and failure load and stiffness were obtained during load-to-failure testing. Failure method was recorded. RESULTS: There was no significant difference between transosseous suture anchor repairs and suture bridge repairs for elongation (4.0 +/- 1.60 mm vs 3.5 +/- 1.1 mm, P = .31), failure load (408 +/- 93 N vs 419 +/- 62 N, P = .70), or stiffness (58 +/- 10 N/mm vs 58 +/- 14 N/mm, P = .94). The most common mode of failure with each method was suture cutting through tendon. CONCLUSION: The suture bridge repair exhibited similar biomechanical performance during cyclic and load-to-failure testing as a transosseous-suture anchor repair, which historically has been performed in open or mini-open fashion. CLINICAL RELEVANCE: Arthroscopic rotator cuff repairs can be performed that are as strong as open or mini-open repairs. PMID- 19556469 TI - The 3-in-1 procedure for recurrent dislocation of the patella in skeletally immature children and adolescents. AB - BACKGROUND: Recurrent patellar dislocations are common injuries in children and adolescents. The subjective and functional results of soft tissue surgical management in a population that was skeletally immature at the time of surgery have not been reported. HYPOTHESIS: The 3-in-1 procedure is an effective treatment for recurrent patellar dislocation in skeletally immature patients. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Twenty-five skeletally immature patients (age at operation, 13.5 +/- 3.8 years) who were practicing sports and suffering from recurrent unilateral patellar dislocation were included in the study and followed until skeletal maturation. Clinical evaluation included the modified Cincinnati rating system and the Kujala score, anthropometry, plain radiography, and isokinetic dynamometry. RESULTS: The average follow-up was 3.8 years (range, 2.5-6 years). The mean modified Cincinnati score increased from 51.7 +/- 12.6 preoperatively to 94.3 +/- 10.8 (P < .02), while the mean Kujala scores increased from 52.4 +/- 12.7 preoperatively to 93.8 +/- 14.2 (P < .02). The Insall-Salvati index remained essentially unchanged, being 1.04 +/- 0.2 preoperatively and 1.02 +/- 0.3 at latest follow-up. Significant differences were found between the operated and the contralateral limb in the various isokinetic strength variables at all angular velocities measured at the latest follow-up (.05 < P < .0042). CONCLUSION: The 3-in-1 procedure is a safe, reliable management option for recurrent patellar dislocation in skeletally immature patients. Side-to-side differences in isokinetic strength and in anthropometric indices persist despite subjective success of the procedure. PMID- 19556471 TI - Incidence of glenohumeral instability in collegiate athletics. AB - BACKGROUND: Glenohumeral instability is a common injury sustained by young athletes. Surprisingly, little is known regarding the incidence of glenohumeral instability in collegiate athletes or the relevant risk factors for injury. A better understanding of the populations most at risk may be used to develop preventive strategies. HYPOTHESIS: The incidence of glenohumeral instability in collegiate athletics is high, and it is affected by sex, sport, type of event, and mechanism of injury. STUDY DESIGN: Descriptive epidemiologic study. METHODS: The National Collegiate Athletic Association injury database was queried for all glenohumeral instability events occurring between the years 1989 and 2004. An analysis of the injuries was performed by sport, activity (competition versus practice), sex, type of event (primary versus recurrent), mechanism of injury, and time loss from athletic performance. Incidence rates and incidence rate ratios were calculated. RESULTS: A total of 4080 glenohumeral instability events were documented for an incidence rate of 0.12 injuries per 1000 athlete exposures. The sport with the greatest injury rate was men's spring football, with 0.40 injuries per 1000 athlete exposures. Overall, athletes sustained more glenohumeral instability events during games than practices (incidence rate ratio [IRR], 3.50; 95% confidence interval [CI], 3.29-3.73). Male athletes sustained more injuries than did female athletes (IRR, 2.67; 95% CI, 2.43-2.93). Female athletes were more likely to sustain an instability event as the result of contact with an object (IRR, 2.43; 95% CI, 2.08-2.84), whereas male athletes were more likely to sustain an event from player contact (IRR, 2.74; 95% CI, 2.31 3.25). Time lost to sport (>10 days) occurred in 45% of glenohumeral instability events. CONCLUSION: Glenohumeral instability is a relatively common injury sustained by collegiate athletes. More injuries occurred during competition and among male athletes. PMID- 19556470 TI - The long-term effect of 2 postoperative rehabilitation programs after anterior cruciate ligament reconstruction: a randomized controlled clinical trial with 2 years of follow-up. AB - BACKGROUND: There is no consensus regarding the optimal postoperative rehabilitation program after anterior cruciate ligament (ACL) reconstruction. PURPOSE: The purpose of this study was to examine the long-term outcome of a 6 month neuromuscular exercise (NE) training program versus a traditional strength exercise (SE) training program after ACL reconstruction. STUDY DESIGN: Randomized controlled trial; Level of evidence, 1. METHODS: Seventy-four patients were randomly assigned to either a NE program or a SE program and tested preoperatively and at 6 months, 1 year, and 2 years after ACL reconstruction. Outcome measurements were as follows: Cincinnati knee score, visual analog scale for pain and global function, Short Form 36, functional knee tests, and isokinetic muscle strength tests. RESULTS: There were no significant differences between the NE and SE programs 1 and 2 years after ACL reconstruction for the primary outcome measurement (Cincinnati knee score). There were significantly improved knee function (global function) and reduced pain during activity for the NE group, compared with the SE group, and significantly improved hamstring muscle strength for the SE group, compared with the NE group, 2 years after ACL reconstruction. CONCLUSION: On the basis of these results, a postoperative program combining both NE and SE should be included after ACL reconstruction to improve knee function. PMID- 19556472 TI - Out with the old, in with the new? PMID- 19556474 TI - Fusion research. ITER gets the nod for slower, step-by-step approach. PMID- 19556473 TI - Biosecurity. Discovery of untracked pathogen vials at army lab sparks concerns. PMID- 19556475 TI - Depression gene. Back to the drawing board for psychiatric genetics. PMID- 19556476 TI - International collaboration. Bioscientists slowly bridge the Taiwan Strait. PMID- 19556477 TI - Science in society. China reins in wilder impulses in treatment of 'Internet addiction'. PMID- 19556479 TI - Biodiversity. Biodiversity databases spread, prompting unification call. PMID- 19556481 TI - The brain collector. PMID- 19556483 TI - Space weather forecasting. Are we ready for the next solar maximum? No way, say scientists. PMID- 19556482 TI - Microbiology. Antibiotics in nature: beyond biological warfare. PMID- 19556484 TI - The global alliance for chronic diseases. PMID- 19556485 TI - Current Brazilian law on animal experimentation. PMID- 19556486 TI - Sex in Leishmania. PMID- 19556487 TI - Creationist beliefs in Europe. PMID- 19556488 TI - Research ethics. The NIH draft guidelines on human stem cell research. PMID- 19556489 TI - Information technology. Innovating education in Croatia. PMID- 19556490 TI - Microbiology. Seeing green and red in diatom genomes. PMID- 19556491 TI - Evolution. Auxin at the evo-devo intersection. PMID- 19556492 TI - Developmental biology. Phase transition in a cell. PMID- 19556493 TI - Chemistry. Rate control and reaction engineering. PMID- 19556494 TI - Computer science. Building an open Cloud. PMID- 19556495 TI - Steps to the clinic. Stem cells. Introduction. PMID- 19556496 TI - FDA regulation of stem cell-based products. AB - Cell self-renewal and the capacity to differentiate into multiple cell types (pluripotency) are biological attributes casting stem cells as attractive candidates for development of therapies targeting indications that involve functional restoration of damaged tissues. In the United States, clinical trials designed to demonstrate the safety and effectiveness of stem cell-based products are regulated by the U.S. Food and Drug Administration (FDA). To ensure that subjects enrolled in a clinical study involving stem cell-based products are not exposed to significant and unreasonable risk, the FDA reviews medical and scientific information that encompasses delineation of product-specific characteristics and preclinical testing to determine whether there is sufficient safety assurance to permit initiation of human clinical studies. PMID- 19556497 TI - Medical innovation versus stem cell tourism. AB - Stem cell tourism is criticized on grounds of consumer fraud, blatant lack of scientific justification, and patient safety. However, the issues are complex because they invoke questions concerning the limits of acceptable medical innovation and medical travel. Here we discuss these issues and articulate conditions under which "unproven" therapies may be offered to patients outside of regular clinical trials. PMID- 19556498 TI - The role of stromal stem cells in tissue regeneration and wound repair. AB - The process of wound repair in epithelium-lined organs of mammals is complex and is influenced by numerous secreted factors including cytokines, growth factors, and chemokines. However, the cellular organizers of this process are still not understood. Recent studies of tissue regeneration in organisms with simpler development have uncovered details about the activity of stem cells in the mesenchyme (the blastema) during this process. These blastemal cells are well positioned to interpret cues from the environment and to execute decisions about the direction of wound repair. In mammalian wounds, stromal stem cells appear to be positioned to perform functions similar to those of blastemal cells, including communication with both the overlying epithelium and the inflammatory cells in the mesenchyme. PMID- 19556499 TI - The increasing complexity of the cancer stem cell paradigm. AB - The investigation and study of cancer stem cells (CSCs) have received enormous attention over the past 5 to 10 years but remain topics of considerable controversy. Opinions about the validity of the CSC hypothesis, the biological properties of CSCs, and the relevance of CSCs to cancer therapy differ widely. In the following commentary, we discuss the nature of the debate, the parameters by which CSCs can or cannot be defined, and the identification of new potential therapeutic targets elucidated by considering cancer as a problem in stem cell biology. PMID- 19556501 TI - Competitive interactions between cells: death, growth, and geography. AB - Competitive interactions between cells are the basis of many homeostatic processes in biology. Some of the best-described cases of competition between cells occur in Drosophila: cell competition, whereby somatic cells within a growing epithelium compete with one another for contribution to the adult, and stem cell competition, in which germline or somatic stem cells vie for residency in the niche. Both types of competition are conserved physiological processes, with much to tell us about how cellular neighborhoods influence cell behavior, and have importance to stem cell biology, regeneration and transplantation, and cancer. PMID- 19556500 TI - Growth factors, matrices, and forces combine and control stem cells. AB - Stem cell fate is influenced by a number of factors and interactions that require robust control for safe and effective regeneration of functional tissue. Coordinated interactions with soluble factors, other cells, and extracellular matrices define a local biochemical and mechanical niche with complex and dynamic regulation that stem cells sense. Decellularized tissue matrices and synthetic polymer niches are being used in the clinic, and they are also beginning to clarify fundamental aspects of how stem cells contribute to homeostasis and repair, for example, at sites of fibrosis. Multifaceted technologies are increasingly required to produce and interrogate cells ex vivo, to build predictive models, and, ultimately, to enhance stem cell integration in vivo for therapeutic benefit. PMID- 19556502 TI - Elevated CO2 enhances otolith growth in young fish. AB - A large fraction of the carbon dioxide added to the atmosphere by human activity enters the sea, causing ocean acidification. We show that otoliths (aragonite ear bones) of young fish grown under high CO2 (low pH) conditions are larger than normal, contrary to expectation. We hypothesize that CO2 moves freely through the epithelium around the otoliths in young fish, accelerating otolith growth while the local pH is controlled. This is the converse of the effect commonly reported for structural biominerals. PMID- 19556503 TI - High-resolution NMR in magnetic fields with unknown spatiotemporal variations. AB - Nuclear magnetic resonance (NMR) experiments are usually carried out in homogeneous magnetic fields. In many cases, however, high-resolution spectra are virtually impossible to obtain because of the inherent heterogeneity of the samples or living organisms under investigation, as well as the poor homogeneity of the magnets (particularly when bulky samples must be placed outside their bores). Unstable power supplies and vibrations arising from cooling can lead to field fluctuations in time as well as space. We show how high-resolution NMR spectra can be obtained in inhomogeneous fields with unknown spatiotemporal variations. Our method, based on coherence transfer between spins, can accommodate spatial inhomogeneities of at least 11 gauss per centimeter and temporal fluctuations slower than 2 hertz. PMID- 19556504 TI - White phosphorus is air-stable within a self-assembled tetrahedral capsule. AB - The air-sensitive nature of white phosphorus underlies its destructive effect as a munition: Tetrahedral P4 molecules readily react with atmospheric dioxygen, leading this form of the element to spontaneously combust upon exposure to air. Here, we show that hydrophobic P4 molecules are rendered air-stable and water soluble within the hydrophobic hollows of self-assembled tetrahedral container molecules, which form in water from simple organic subcomponents and iron(II) ions. This stabilization is not achieved through hermetic exclusion of O2 but rather by constriction of individual P4 molecules; the addition of oxygen atoms to P4 would result in the formation of oxidized species too large for their containers. The phosphorus can be released in controlled fashion without disrupting the cage by adding the competing guest benzene. PMID- 19556505 TI - Trapping molecules on a chip. AB - Magnetic trapping of atoms on chips has recently become straightforward, but analogous trapping of molecules has proved to be challenging. We demonstrated trapping of carbon monoxide molecules above a chip using direct loading from a supersonic beam. Upon arrival above the chip, the molecules are confined in tubular electric field traps approximately 20 micrometers in diameter, centered 25 micrometers above the chip, that move with the molecular beam at a velocity of several hundred meters per second. An array of these miniaturized moving traps is brought to a standstill over a distance of only a few centimeters. After a certain holding time, the molecules are accelerated off the chip again for detection. This loading and detection methodology is applicable to a wide variety of polar molecules, enabling the creation of a gas-phase molecular laboratory on a chip. PMID- 19556506 TI - Postmating sexual selection favors males that sire offspring with low fitness. AB - Despite the costs of mating, females of most taxa mate with multiple males. Polyandrous females are hypothesized to gain genetic benefits for their offspring, but this assumes paternity bias favoring male genotypes that enhance offspring viability. We determined net male genetic effects on female and offspring fitness in a seed beetle and then tested whether fertilization success was biased in favor of high-quality male genotypes in double mating experiments. Contrary to expectations, high-quality male genotypes consistently had a lower postmating fertilization success in two independent assays. Our results imply that sexually antagonistic adaptations have a major and unappreciated influence on male postmating fertilization success. Such genetic variation renders indirect genetic benefits an unlikely driver of the evolution of polyandry. PMID- 19556507 TI - Dynamic signaling network for the specification of embryonic pancreas and liver progenitors. AB - Studies of the formation of pancreas and liver progenitors have focused on individual inductive signals and cellular responses. Here, we investigated how bone morphogenetic protein, transforming growth factor-beta (TGFbeta), and fibroblast growth factor signaling pathways converge on the earliest genes that elicit pancreas and liver induction in mouse embryos. The inductive network was found to be dynamic; it changed within hours. Different signals functioned in parallel to induce different early genes, and two permutations of signals induced liver progenitor domains, which revealed flexibility in cell programming. Also, the specification of pancreas and liver progenitors was restricted by the TGFbeta pathway. These findings may enhance progenitor cell specification from stem cells for biomedical purposes and can help explain incomplete programming in stem cell differentiation protocols. PMID- 19556508 TI - Mitochondrial STAT3 supports Ras-dependent oncogenic transformation. AB - Signal transducer and activator of transcription 3 (STAT3) is a latent cytoplasmic transcription factor responsive to cytokine signaling and tyrosine kinase oncoproteins by nuclear translocation when it is tyrosine-phosphorylated. We report that malignant transformation by activated Ras is impaired without STAT3, in spite of the inability of Ras to drive STAT3 tyrosine phosphorylation or nuclear translocation. Moreover, STAT3 mutants that cannot be tyrosine phosphorylated, that are retained in the cytoplasm, or that cannot bind DNA nonetheless supported Ras-mediated transformation. Unexpectedly, STAT3 was detected within mitochondria, and exclusive targeting of STAT3 to mitochondria without nuclear accumulation facilitated Ras transformation. Mitochondrial STAT3 sustained altered glycolytic and oxidative phosphorylation activities characteristic of cancer cells. Thus, in addition to its nuclear transcriptional role, STAT3 regulates a metabolic function in mitochondria, supporting Ras dependent malignant transformation. PMID- 19556509 TI - Synthetic heterochromatin bypasses RNAi and centromeric repeats to establish functional centromeres. AB - In the central domain of fission yeast centromeres, the kinetochore is assembled on CENP-A(Cnp1) nucleosomes. Normally, small interfering RNAs generated from flanking outer repeat transcripts direct histone H3 lysine 9 methyltransferase Clr4 to homologous loci to form heterochromatin. Outer repeats, RNA interference (RNAi), and centromeric heterochromatin are required to establish CENP-A(Cnp1) chromatin. We demonstrated that tethering Clr4 via DNA-binding sites at euchromatic loci induces heterochromatin assembly, with or without active RNAi. This synthetic heterochromatin completely substitutes for outer repeats on plasmid-based minichromosomes, promoting de novo CENP-A(Cnp1) and kinetochore assembly, to allow their mitotic segregation, even with RNAi inactive. Thus, the role of outer repeats in centromere establishment is simply the provision of RNAi substrates to direct heterochromatin formation; H3K9 methylation-dependent heterochromatin is alone sufficient to form functional centromeres. PMID- 19556510 TI - Genomic footprints of a cryptic plastid endosymbiosis in diatoms. AB - Diatoms and other chromalveolates are among the dominant phytoplankters in the world's oceans. Endosymbiosis was essential to the success of chromalveolates, and it appears that the ancestral plastid in this group had a red algal origin via an ancient secondary endosymbiosis. However, recent analyses have turned up a handful of nuclear genes in chromalveolates that are of green algal derivation. Using a genome-wide approach to estimate the "green" contribution to diatoms, we identified >1700 green gene transfers, constituting 16% of the diatom nuclear coding potential. These genes were probably introduced into diatoms and other chromalveolates from a cryptic endosymbiont related to prasinophyte-like green algae. Chromalveolates appear to have recruited genes from the two major existing algal groups to forge a highly successful, species-rich protist lineage. PMID- 19556511 TI - Solution nuclear magnetic resonance structure of membrane-integral diacylglycerol kinase. AB - Escherichia coli diacylglycerol kinase (DAGK) represents a family of integral membrane enzymes that is unrelated to all other phosphotransferases. We have determined the three-dimensional structure of the DAGK homotrimer with the use of solution nuclear magnetic resonance. The third transmembrane helix from each subunit is domain-swapped with the first and second transmembrane segments from an adjacent subunit. Each of DAGK's three active sites resembles a portico. The cornice of the portico appears to be the determinant of DAGK's lipid substrate specificity and overhangs the site of phosphoryl transfer near the water-membrane interface. Mutations to cysteine that caused severe misfolding were located in or near the active site, indicating a high degree of overlap between sites responsible for folding and for catalysis. PMID- 19556513 TI - In situ analysis of protein S-glutathionylation in lung tissue using glutaredoxin 1-catalyzed cysteine derivatization. AB - Protein S-glutathionylation (PSSG) is a posttranslational modification that involves the conjugation of the small antioxidant molecule glutathione to cysteine residues and is emerging as a critical mechanism of redox-based signaling. PSSG levels increase under conditions of oxidative stress and are controlled by glutaredoxins (Grx) that, under physiological conditions, preferentially deglutathionylate cysteines and restore sulfhydryls. Both the occurrence and distribution of PSSG in tissues is unknown because of the labile nature of this oxidative event and the lack of specific reagents. The goal of this study was to establish and validate a protocol that enables detection of PSSG in situ, using the property of Grx to deglutathionylate cysteines. Using Grx1-catalyzed cysteine derivatization, we evaluated PSSG content in mice subjected to various models of lung injury and fibrosis. In control mice, PSSG was detectable primarily in the airway epithelium and alveolar macrophages. Exposure of mice to NO(2) resulted in enhanced PSSG levels in parenchymal regions, while exposure to O(2) resulted in minor detectable changes. Finally, bleomycin exposure resulted in marked increases in PSSG reactivity both in the bronchial epithelium as well as in parenchymal regions. Taken together, these findings demonstrate that Grx1-based cysteine derivatization is a powerful technique to specifically detect patterns of PSSG expression in lungs, and will enable investigations into regional changes in PSSG content in a variety of diseases. PMID- 19556514 TI - Independent effects of intra- and extracellular Abeta on learning-related gene expression. AB - Alzheimer's disease is characterized by numerous pathological abnormalities, including amyloid beta (Abeta) deposition in the brain parenchyma and vasculature. In addition, intracellular Abeta accumulation may affect neuronal viability and function. In this study, we evaluated the effects of different forms of Abeta on cognitive decline by analyzing the behavioral induction of the learning-related gene Arc/Arg3.1 in three different transgenic mouse models of cerebral amyloidosis (APPPS1, APPDutch, and APP23). Following a controlled spatial exploration paradigm, reductions in both the number of Arc-activated neurons and the levels of Arc mRNA were seen in the neocortices of depositing mice from all transgenic lines (deficits ranging from 14 to 26%), indicating an impairment in neuronal encoding and network activation. Young APPDutch and APP23 mice exhibited intracellular, granular Abeta staining that was most prominent in the large pyramidal cells of cortical layer V; these animals also had reductions in levels of Arc. In the dentate gyrus, striking reductions (up to 58% in aged APPPS1 mice) in the number of Arc-activated cells were found. Single-cell analyses revealed both the proximity to fibrillar amyloid in aged mice, and the transient presence of intracellular granular Abeta in young mice, as independent factors that contribute to reduced Arc levels. These results provide evidence that two independent Abeta pathologies converge in their impact on cognitive function in Alzheimer's disease. PMID- 19556515 TI - Gastrin is an essential cofactor for helicobacter-associated gastric corpus carcinogenesis in C57BL/6 mice. AB - We have previously described a synergistic interaction between hypergastrinemia and Helicobacter felis infection on gastric corpus carcinogenesis in FVB/N mice housed under specific-pathogen-free conditions. However, gastrin-deficient (GAS KO) mice on a mixed C57BL/6/129Sv genetic background maintained in conventional housing were reported to develop spontaneous gastric antral tumors. Therefore, we investigated the role of gastrin in Helicobacter-associated gastric carcinogenesis in H. felis-infected mice on a uniform C57BL/6 background housed in specific-pathogen-free conditions. Hypergastrinemic transgenic (INS-GAS) mice, GAS-KO mice, and C57BL/6 wild-type mice were infected with H. felis for either 12 or 18 months. At 12 months postinfection, INS-GAS mice had mild corpus dysplasia, while B6 wild-type mice had either severe gastritis or metaplasia, and GAS-KO mice had only mild to moderate gastritis. At 18 months postinfection, both INS GAS and B6 wild-type mice had both severe atrophic gastritis and corpus dysplasia, while GAS-KO mice had severe gastritis with mild gastric atrophy, but no corpus dysplasia. In contrast, both GAS-KO and B6 wild-type mice had mild to moderate antral dysplasia, while INS-GAS mice did not. H. felis antral colonization remained stable over time among the three groups of mice. These results point to a distinct effect of gastrin on carcinogenesis of both the gastric corpus and antrum, suggesting that gastrin is an essential cofactor for gastric corpus carcinogenesis in C57BL/6 mice. PMID- 19556516 TI - Epstein-barr virus-induced gene-3 is expressed in human atheroma plaques. AB - Atherosclerosis is characterized by a complex immune response in the vessel wall, involving both inflammation and autoimmune processes. Epstein-Barr virus-induced gene 3 (Ebi3) is a member of the interleukin (IL)-12 heterodimeric cytokine family, which has important immunomodulatory functions. To date, little is known about the role of Ebi3 in vascular disease. We examined the expression of Ebi3 in human atheromatous lesions and analyzed its transcriptional regulation in vascular cells. The in situ expression of Ebi3 in human endarterectomy specimens was analyzed by immunohistochemistry. In these lesions, smooth muscle cells expressed Ebi3 as well as the IL-27alpha/p28 and IL-12alpha/p35 subunits. Primary aortic smooth muscle cells up-regulated Ebi3 in response to proinflammatory stimuli like tumor necrosis factor-alpha and interferon-gamma. Interestingly, pretreatment of these cells with the peroxisome proliferator-activated receptor gamma agonist rosiglitazone strongly reduced Ebi3 induction. Chromatin immunoprecipitation experiments revealed that this inhibition is due to interference with p65/RelA recruitment to the Ebi3 promoter. Our data support a possible role of Ebi3 in atherogenesis either as homodimer or as IL-27/IL-35 heterodimer, and suggest that Ebi3 could be an interesting target for therapeutic manipulation in atherosclerosis. PMID- 19556517 TI - Superior immune response to protein-conjugate versus free pneumococcal polysaccharide vaccine in chronic obstructive pulmonary disease. AB - RATIONALE: Debate exists about the immunogenicity and protective efficacy of antibodies produced by the 23-valent pneumococcal polysaccharide vaccine (PPSV23) in chronic obstructive pulmonary disease (COPD). The 7-valent diphtheria conjugated pneumococcal polysaccharide vaccine (PCV7) induces a more robust immune response than PPSV23 in healthy elderly adults. OBJECTIVES: We hypothesized that serotype-specific IgG antibody concentration and functional antibody activity would be superior after PCV7 vaccination compared with PPSV23 in moderate to severe COPD. We also posited that older age and prior PPSV23 vaccination would be associated with reduced vaccine responsiveness. METHODS: One hundred twenty patients with COPD were randomized to PPSV23 (63 subjects) or PCV7 (57 subjects). IgG concentrations were determined by ELISA; functional antibody activity was assayed with a standardized opsonophagocytosis assay and reported as an opsonization killing index (OPK). Increases in serotype-specific IgG and OPK at 1 month post vaccination were compared within and between vaccine groups. MEASUREMENTS AND MAIN RESULTS: Both vaccines were well tolerated. Within each study group, postvaccination IgG and OPK were higher than baseline (P < 0.01) for all serotypes. Adjusted for baseline levels, postvaccination IgG was higher in the PCV7 group than the PPSV23 group for all seven serotypes, reaching statistical significance for five (P < 0.05). PCV7 resulted in a higher OPK for six of seven serotypes (statistically greater for four) compared with PPSV23. In multivariate analyses, younger age, vaccine naivety, and receipt of PCV7 were associated with increased OPK responses. CONCLUSIONS: PCV7 induces a superior immune response at 1 month post vaccination compared with PPSV23 in COPD. Older age and prior PPSV23 reduce vaccine responsiveness. Clinical trial registered with www.clinicaltrials.gov (NCT00457977). PMID- 19556519 TI - Improved biomass stove intervention in rural Mexico: impact on the respiratory health of women. AB - RATIONALE: Exposure to biomass smoke has been related to adverse health effects. In Mexico, one household in four still cooks with biomass fuel, but there has been no evaluation of the health impact of reducing indoor air pollution. OBJECTIVES: To evaluate the health impact of the introduction of an improved biomass stove (Patsari; Interdisciplinary Group for Appropriate Rural Technology [GIRA], Patzcuaro, Mexico) in Mexican women. METHODS: A randomized controlled trial was conducted in the Central Mexican state of Michoacan. Households were randomized to receive the Patsari stove or keep their traditional open fire. A total of 552 women were followed with monthly visits over 10 months to assess stove use, inquire about respiratory and other symptoms, and obtain lung function measurements. Statistical analysis was conducted using longitudinal models. MEASUREMENTS AND MAIN RESULTS: Adherence to the intervention was low (50%). Women who reported using the Patsari stove most of the time compared with those using the open fire had significantly lower risk of respiratory symptoms (relative risk [RR], 0.77; 95% confidence interval [CI], 0.62-0.95 for cough and RR, 0.29; 95% CI, 0.11-0.77 for wheezing) adjusted for confounders. Similar results were found for other respiratory symptoms as well as for eye discomfort, headache, and back pain. Actual use of the Patsari stove was associated with a lower FEV(1) decline (31 ml) compared with the open fire use (62 ml) over 1 year of follow-up (P = 0.012) for women 20 years of age and older, adjusting for confounders. CONCLUSIONS: The use of the Patsari stove was significantly associated with a reduction of symptoms and of lung function decline comparable to smoking cessation. PMID- 19556518 TI - Contribution of epithelial-derived fibroblasts to bleomycin-induced lung fibrosis. AB - RATIONALE: Lung fibroblasts are key mediators of fibrosis resulting in accumulation of excessive interstitial collagen and extracellular matrix, but their origins are not well defined. OBJECTIVES: We aimed to elucidate the contribution of lung epithelium-derived fibroblasts via epithelial-mesenchymal transition (EMT) in the intratracheal bleomycin model. METHODS: Primary type II alveolar epithelial cells were cultured from Immortomice and exposed to transforming growth factor-beta(1) and epidermal growth factor. Cell fate reporter mice that permanently mark cells of lung epithelial lineage with beta galactosidase were developed to study EMT, and bone marrow chimeras expressing green fluorescent protein under the control of the fibroblast-associated S100A4 promoter were generated to examine bone marrow-derived fibroblasts. Mice were given intratracheal bleomycin (0.08 unit). Immunostaining was performed for S100A4, beta-galactosidase, green fluorescent protein, and alpha-smooth muscle actin. MEASUREMENTS AND MAIN RESULTS: In vitro, primary type II alveolar epithelial cells undergo phenotypic changes of EMT when exposed to transforming growth factor-beta(1) and epidermal growth factor with loss of prosurfactant protein C and E-cadherin and gain of S100A4 and type I procollagen. In vivo, using cell fate reporter mice, approximately one-third of S100A4-positive fibroblasts were derived from lung epithelium 2 weeks after bleomycin administration. From bone marrow chimera studies, one-fifth of S100A4-positive fibroblasts were derived from bone marrow at this same time point. Myofibroblasts rarely derived from EMT or bone marrow progenitors. CONCLUSIONS: Both EMT and bone marrow progenitors contribute to S100A4-positive fibroblasts in bleomycin induced lung fibrosis. However, neither origin is a principal contributor to lung myofibroblasts. PMID- 19556520 TI - Novel pharmacophores of connexin43 based on the "RXP" series of Cx43-binding peptides. AB - Gap junction pharmacology is a nascent field. Previous studies have identified molecules that enhance intercellular communication, and may offer potential for innovative antiarrhythmic therapy. However, their specific molecular target(s) and mechanism(s) of action remain unknown. Previously, we identified a 34-aa peptide (RXP-E) that binds the carboxyl terminal domain of Cx43 (Cx43CT) and prevents cardiac gap junction closure and action potential propagation block. These results supported the feasibility of a peptide-based pharmacology to Cx43, but the structure of the core active element in RXP-E, an essential step for pharmacological development, remained undefined. Here, we used a combination of molecular modeling, surface plasmon resonance, nuclear magnetic resonance and patch-clamp strategies to define, for the first time, a unique ensemble of pharmacophores that bind Cx43CT and prevent closure of Cx43 channels. Two particular molecules are best representatives of this family: a cyclized heptapeptide (called CyRP-71) and a linear octapeptide of sequence RRNYRRNY. These 2 small compounds offer the first structural platform for the design of Cx43-interacting gap junction openers. Moreover, the structure of these compounds offers an imprint of a region of Cx43CT that is fundamental to gap junction channel function. PMID- 19556521 TI - Protein kinase C{alpha}, but not PKC{beta} or PKC{gamma}, regulates contractility and heart failure susceptibility: implications for ruboxistaurin as a novel therapeutic approach. AB - Protein kinase (PK)Calpha, PKCbeta, and PKCgamma comprise the conventional PKC isoform subfamily, which is thought to regulate cardiac disease responsiveness. Indeed, mice lacking the gene for PKCalpha show enhanced cardiac contractility and reduced susceptibility to heart failure. Recent data also suggest that inhibition of conventional PKC isoforms with Ro-32-0432 or Ro-31-8220 enhances heart function and antagonizes failure, although the isoform responsible for these effects is unknown. Here, we investigated mice lacking PKCalpha, PKCbeta, and PKCgamma for effects on cardiac contractility and heart failure susceptibility. PKCalpha(-/-) mice, but not PKCbetagamma(-/-) mice, showed increased cardiac contractility, myocyte cellular contractility, Ca(2+) transients, and sarcoplasmic reticulum Ca(2+) load. PKCalpha(-/-) mice were less susceptible to heart failure following long-term pressure-overload stimulation or 4 weeks after myocardial infarction injury, whereas PKCbetagamma(-/-) mice showed more severe failure. Infusion of ruboxistaurin (LY333531), an orally available PKCalpha/beta/gamma inhibitor, increased cardiac contractility in wild-type and PKCbetagamma(-/-) mice, but not in PKCalpha(-/-) mice. More importantly, ruboxistaurin prevented death in wild-type mice throughout 10 weeks of pressure overload stimulation, reduced ventricular dilation, enhanced ventricular performance, reduced fibrosis, and reduced pulmonary edema comparable to or better than metoprolol treatment. Ruboxistaurin was also administered to PKCbetagamma(-/-) mice subjected to pressure overload, resulting in less death and heart failure, implicating PKCalpha as the primary target of this drug in mitigating heart disease. As an aside, PKCalphabetagamma triple-null mice showed no defect in cardiac hypertrophy following pressure-overload stimulation. In conclusion, PKCalpha functions distinctly from PKCbeta and PKCgamma in regulating cardiac contractility and heart failure, and broad-acting PKC inhibitors such as ruboxistaurin could represent a novel therapeutic approach in treating human heart failure. PMID- 19556522 TI - Palmitoylation of ATP-binding cassette transporter A1 is essential for its trafficking and function. AB - ATP-binding cassette transporter (ABC)A1 lipidates apolipoprotein A-I both directly at the plasma membrane and also uses lipids from the late endosomal or lysosomal compartment in the internal lipidation of apolipoprotein A-I. However, how ABCA1 targeting to these specific membranes is regulated remains unknown. Palmitoylation is a dynamically regulated lipid modification that targets many proteins to specific membrane domains. We hypothesized that palmitoylation may also regulate ABCA1 transport and function. Indeed, ABCA1 is robustly palmitoylated at cysteines 3, -23, -1110, and -1111. Abrogation of palmitoylation of ABCA1 by mutation of the cysteines results in a reduction of ABCA1 localization at the plasma membranes and a reduction in the ability of ABCA1 to efflux lipids to apolipoprotein A-I. ABCA1 is palmitoylated by the palmitoyl transferase DHHC8, and increasing DHHC8 protein results in increased ABCA1 mediated lipid efflux. Thus, palmitoylation regulates ABCA1 localization at the plasma membrane, and regulates its lipid efflux ability. PMID- 19556523 TI - Aggregated LDL in contact with macrophages induces local increases in free cholesterol levels that regulate local actin polymerization. AB - OBJECTIVE: Interaction of macrophages with aggregated matrix-anchored lipoprotein deposits is an important initial step in atherogenesis. Aggregated lipoproteins require different cellular uptake processes than those used for endocytosis of monomeric lipoproteins. In this study, we tested the hypothesis that engagement of aggregated LDL (agLDL) by macrophages could lead to local increases in free cholesterol levels and that these increases in free cholesterol regulate signals that control cellular actin. METHODS AND RESULTS: AgLDL resides for prolonged periods in surface-connected compartments. Although agLDL is still extracellular, we demonstrate that an increase in free cholesterol occurs at sites of contact between agLDL and cells because of hydrolysis of agLDL-derived cholesteryl ester. This increase in free cholesterol causes enhanced actin polymerization around the agLDL. Inhibition of cholesteryl ester hydrolysis results in decreased actin polymerization. CONCLUSIONS: We describe a novel process that occurs during agLDL macrophage interactions in which local release of free cholesterol causes local actin polymerization, promoting a pathological positive feedback loop for increased catabolism of agLDL and eventual foam cell formation. PMID- 19556524 TI - Cadmium is a novel and independent risk factor for early atherosclerosis mechanisms and in vivo relevance. AB - OBJECTIVE: Although cadmium (Cd) is an important and common environmental pollutant and has been linked to cardiovascular diseases, little is known about its effects in initial stages of atherosclerosis. METHODS AND RESULTS: In the 195 young healthy women of the Atherosclerosis Risk Factors in Female Youngsters (ARFY) study, cadmium (Cd) level was independently associated with early atherosclerotic vessel wall thickening (intima-media thickness exceeding the 90th percentile of the distribution; multivariable OR 1.6[1.1.-2.3], P=0.016). In line, Cd-fed ApoE knockout mice yielded a significantly increased aortic plaque surface compared to controls (9.5 versus 26.0 mm(2), P<0.004). In vitro results indicate that physiological doses of Cd increase vascular endothelial permeability up to 6-fold by (1) inhibition of endothelial cell proliferation, and (2) induction of a caspase-independent but Bcl-xL-inhibitable form of cell death more than 72 hours after Cd addition. Both phenomena are preceded by Cd induced DNA strand breaks and a cellular DNA damage response. Zinc showed a potent protective effect against deleterious effects of Cd both in the in vitro and human studies. CONCLUSIONS: Our research suggests Cd has promoting effects on early human and murine atherosclerosis, which were partly offset by high Zn concentrations. PMID- 19556525 TI - Binge drinking and mortality from all causes and cerebrovascular diseases in korean men and women: a Kangwha cohort study. AB - BACKGROUND AND PURPOSE: The purpose of this study was to examine the association between binge drinking and risks of mortality due to all causes of death with a focus on cerebrovascular disease in Korean men and women. METHODS: This study followed a cohort of 6291 residents in Kangwha County, aged > or =55 years in March 1985, for their cause-specific mortality for 20.8 years up to December 31, 2005. We calculated hazard ratio of mortality by experience or frequency of binge drinking using the Cox proportional hazard model. Binge drinking was defined as having > or =6 drinks on one occasion. RESULTS: In men, binge drinkers who drink daily had an increased risk of mortality from all causes (hazard ratio, 1.33; 95% CI, 1.11 to 1.60) as compared with nondrinkers. They showed much increased risks of mortality from total stroke (hazard ratio, 1.86; 95% CI, 1.16 to 2.99) and hemorrhagic stroke (hazard ratio, 3.39; 95% CI, 1.38 to 8.35). Female binge drinkers also showed an increased risk of mortality from cardiovascular disease as compared with female nondrinkers, but the outcome was not statistically significant. CONCLUSIONS: The results of this study suggest that frequent binge drinking has a harmful effect on hemorrhagic stroke in Korean men. These findings need to be confirmed in further studies. PMID- 19556527 TI - Complex traditional Chinese medicine for poststroke motor dysfunction: a systematic review. AB - BACKGROUND AND PURPOSE: For its current dimensions, stroke represents the world's primary health challenge. In China stroke is the second most common cause of death. Traditional Chinese Medicine (TCM) has for many centuries been used, and it is still widely used today in countries of south and east Asia for the treatment of people with stroke. The objective of this systematic review was to evaluate whether complex Traditional Chinese Medicine (cTCM) improves poststroke motor recovery. In particular, we defined cTCM as intervention that included at least acupuncture and Chinese herbal medicine. METHODS: An extensive search including PubMed, EMBASE, CBM, and the Cochrane Library was performed up to December 2007. Randomized clinical trials (RCTs) about cTCM for motor dysfunction of poststroke were searched irrespective of any language. The quality of each trial was assessed according to the Cochrane Reviewers' Handbook 4.2.6. RESULTS: After selection of 11 234 articles, 34 RCTs and quasi-RCTs were included. All these trials were conducted in China and published on Chinese journals. All trials but one reported results in favor of cTCM treatments suggesting a strong publication bias. Because of the significant clinical and methodological heterogeneity, no meta-analysis was performed and thus no cumulative result was obtained pooling data of RCTs. CONCLUSIONS: What appears from this systematic review is that scant data are available to evaluate efficacy of cTCM for poststroke motor dysfunction. Most of the primary studies available for this review were inadequately designed trials characterized by unknown dropout rates and definitional vagueness in outcomes measures. None of the studies approached important end points like death, survival times, rate of dependency, reduction in length of stay in hospital, etc. The key to lead to evidence-based practices is establishing a consensus on standardized relevant outcome measures and then designing and conducting appropriate RCTs that adopt those standards. PMID- 19556526 TI - Walking after stroke: what does treadmill training with body weight support add to overground gait training in patients early after stroke?: a single-blind, randomized, controlled trial. AB - BACKGROUND AND PURPOSE: This study aimed to assess the effectiveness of gait training using body weight support on a treadmill compared with conventional gait training for people with subacute stroke who were unable to walk. METHODS: This was a single-blind, randomized, controlled trial with a 6-month follow-up. Ninety seven subjects were recruited within 6 weeks of stroke onset and were randomly assigned to conventional rehabilitative treatment plus gait training with body weight support on a treadmill (experimental group; n=52) and conventional treatment with overground gait training only (control group; n=45). All subjects were treated in 60-minute sessions every weekday for 4 weeks. Outcome measures were Motricity Index, Trunk Control test, Barthel Index, Functional Ambulation Categories, 10-meter and 6-minute Walk Tests, and Walking Handicap Scale. Assessments were made at baseline, after 20 sessions of treatment, 2 weeks after treatment, and 6 months after stroke. RESULTS: After treatment, all patients were able to walk. Both groups showed improvement in all outcome measures (P<0.0063) at the end of the treatment and at follow-up. No differences were seen between the 2 groups before, during, and after treatment and at follow-up. CONCLUSIONS: In subacute patients with stroke, gait training on a treadmill with body weight support is feasible and as effective as conventional gait training. However, the need for more personnel for treadmill training makes the use of robotically assisted systems more compelling. PMID- 19556528 TI - Reproducibility of fibrous cap status assessment of carotid artery plaques by contrast-enhanced MRI. AB - BACKGROUND AND PURPOSE: Reproducibility in identifying the fibrous cap (FC) of carotid artery plaques by noncontrast-enhanced MRI has been shown to be poor. The objective of this study was to assess the reproducibility of multisequence MRI, including contrast-enhanced images, in assessing FC status. METHODS: Forty-five symptomatic patients with 30% to 69% carotid artery stenosis underwent a multisequence MRI protocol, which included contrast-enhanced images. FC status (ie, discrimination between fibrotic and/or calcified plaques, plaques with a lipid-rich necrotic core and an intact and thick FC, and plaques with a lipid rich necrotic core and a thin and/or ruptured FC) was independently assessed by 3 observers of which one also scored all images on a different occasion. Linear weighted kappa coefficients (kappa) were calculated as indicators of inter- and intraobserver agreement. RESULTS: On a per-slice basis, interobserver agreement was good (kappa=0.60, 0.64, and 0.71), whereas intraobserver agreement was very good (kappa=0.86). On a per-plaque basis, interobserver agreement was good (kappa=0.64, 0.69, and 0.78), whereas intraobserver agreement was very good (kappa=0.96). CONCLUSIONS: This study found good interobserver and very good intraobserver agreement in assessing FC status of carotid artery plaques. Future studies are warranted to determine the predictive value of FC status assessment by multisequence MRI, including contrast-enhanced images, on the occurrence of (recurrent) cerebral ischemic events. PMID- 19556530 TI - Descriptive analysis of the Boston criteria applied to a Dutch-type cerebral amyloid angiopathy population. AB - BACKGROUND AND PURPOSE: Validation of the Boston criteria for the in vivo diagnosis of cerebral amyloid angiopathy (CAA) is challenging, because noninvasive diagnostic tests do not exist. Hereditary cerebral hemorrhage with amyloidosis-Dutch type is an accepted monogenetic model of CAA and diagnosis can be made with certainty based on DNA analysis. The aim of this study was to analyze and refine the existing Boston criteria in patients with hereditary cerebral hemorrhage with amyloidosis-Dutch type. METHODS: We performed T2* weighted MRI in 27 patients with hereditary cerebral hemorrhage with amyloidosis Dutch type to assess the presence and location of microbleeds, intracranial hemorrhages, and superficial siderosis. Using the Boston criteria, subjects were categorized as having: no hemorrhages, possible CAA, probable CAA, and hemorrhagic lesions not qualifying for CAA. The sensitivity of the Boston criteria was calculated separately using intracranial hemorrhages only and using intracranial hemorrhages and microbleeds. RESULTS: The sensitivity of the Boston criteria for probable CAA increased from 48% to 63% when microbleeds were included. For symptomatic subjects only, the sensitivity was 100%. No hemorrhages were identified in the deep white matter, basal ganglia, thalamus, or brainstem. Superficial siderosis, observed in 6 patients, did not increase the sensitivity of the Boston criteria in our study group. CONCLUSIONS: Our data show that using T2*-weighted MRI and including microbleeds increase the sensitivity of the Boston criteria. The exclusion of hemorrhages in the deep white matter, basal ganglia, thalamus, and brainstem does not lower the sensitivity of the Boston criteria. PMID- 19556531 TI - Large-cohort comparison between three-dimensional time-of-flight magnetic resonance and rotational digital subtraction angiographies in intracranial aneurysm detection. AB - BACKGROUND AND PURPOSE: The value of MR angiography varies in diagnosis of intracranial aneurysms due to the difference of equipment and imaging technique. This study was to compare the effectiveness of 3-dimensional time-of-flight MR angiography at 3 T and rotational digital subtraction angiography, both with volume rendering (VR), in detecting intracranial aneurysms. METHODS: One hundred thirty-eight patients with suspected or known aneurysms and other cerebral vascular diseases detected by MR angiography underwent digital subtraction angiography examinations. Postprocessing techniques, including VR and the single artery highlighting method, were performed by a 3-dimensional specialist. The VR digital subtraction angiography was obtained as the gold standard. RESULTS: The rotational digital subtraction angiography and VR-digital subtraction angiography revealed 146 aneurysms in 122 patients and no aneurysms in 16 patients. Of the 276 vessels examined, 136 vessels had 146 aneurysms and 140 vessels had none. Per vessel and per aneurysm sensitivities were 100%, whereas the per vessel accuracy ranged from 97.5% to 98.6% and the per aneurysm accuracy ranged from 95.1% to 97.0%. CONCLUSIONS: VR 3-dimensional time-of-flight MR angiography at 3 T has excellent sensitivity, accuracy, and correlation with VR-digital subtraction angiography and is comparable to catheter cerebral angiography for the evaluation of patients with intracranial aneurysms who tolerate MR angiography well. PMID- 19556529 TI - Plasma and brain matrix metalloproteinase-9 after acute focal cerebral ischemia in rats. AB - BACKGROUND AND PURPOSE: Plasma levels of matrix metalloproteinase-9 (MMP-9) have been proposed to be a useful biomarker for assessing pathological events in brain. Here, we examined the temporal profiles of MMP-9 in blood and brain using a rat model of acute focal cerebral ischemia. METHODS: Plasma and brain levels of MMP-2 and MMP-9 were quantified at 3, 6, 12, and 24 hours after permanent middle cerebral artery occlusion in male Sprague-Dawley rats. Infarct volumes at 24 hours were confirmed with 2,3,5-triphenyl-tetrazolium-chloride staining. RESULTS: In plasma, zymographic bands were detected between 70 and 95 kDa corresponding to pro-MMP-2, pro-MMP-9, and activated MMP-9. A higher 135-kDa band was also seen that is likely to be NGAL-conjugated MMP-9. After ischemia, there were no significant changes in pro-MMP-2, but plasma levels of pro-MMP-9 steadily increased over the course of 24 hours. Activated MMP-9 levels in plasma were significantly elevated only at 24 hours. Plasma NGAL-MMP-9 complexes showed a transient elevation between 3 to 6 hours, after which levels decreased back down to pre-ischemic baselines. In brain homogenates, pro-MMP-2, pro-MMP-9, and activated MMP-9 were seen but no NGAL-MMP-9 bands were detected. Compared to the contralateral hemisphere, MMP-2 and MMP-9 levels in ischemic brain progressively increased over the course of 24 hours. Overall levels of MMP-9 in plasma and brain were significantly correlated, especially at 24 hours. Plasma levels of pro MMP-9 at 24 hours were correlated with final infarct volumes. CONCLUSIONS: Elevated plasma levels of MMP-9 appear to be correlated with brain levels within 24 hours of acute cerebral ischemia in rats. Further investigation into clinical profiles of MMP-9 in acute stroke patients may be useful. PMID- 19556532 TI - M1 susceptibility vessel sign on T2* as a strong predictor for no early recanalization after IV-t-PA in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: In acute stroke patients treated with intravenous tissue plasminogen activator (t-PA), early recanalization of occluded arteries can improve the clinical outcome. The magnetic susceptibility effect of deoxygenated hemoglobin in red thrombi can present as hypointense signals on T2*-weighted gradient echo imaging. We investigated whether the gradient echo imaging M1 susceptibility vessel sign (M1 SVS) can predict no early recanalization after t PA infusion. METHODS: Patients with internal carotid artery and M1 occlusion were prospectively studied. MRI studies, including DWI, T2*, and MRA, were performed before and within 30 minutes and 24 hours after t-PA infusion. The NIHSS score was obtained before and 7 days after t-PA administration. The relationship between the presence of the M1 SVS and no early recanalization and patient outcome was examined. RESULTS: A total of 48 patients (29 men; mean age, 74.6+/ 11.2 years) were enrolled. M1 SVS was present in 13 (27.1%) patients and absent in 35 (72.9%) patients. There were no significant differences in clinical characteristics between the 2 groups. Follow-up MRA within 30 minutes after t-PA infusion revealed that 20 (57.1%) of the 35 patients without the M1 SVS had early recanalization, but that none of the 13 patients with the M1 SVS had early recanalization (P=0.0002). Seven days after t-PA infusion, dramatic improvement was more frequently observed in patients without the M1 SVS (51.4%) than in those with the M1 SVS (0%, P=0.0007). CONCLUSIONS: The M1 SVS on T2* appears to be a strong predictor for no early recanalization after t-PA therapy. PMID- 19556533 TI - Patients enrolled in large randomized clinical trials of antiplatelet treatment for prevention after transient ischemic attack or ischemic stroke are not representative of patients in clinical practice: the Netherlands Stroke Survey. AB - BACKGROUND AND PURPOSE: Many randomized clinical trials have evaluated the benefit of long-term use of antiplatelet drugs in reducing the risk of new vascular events in patients with a recent transient ischemic attack or ischemic stroke. Evidence from these trials forms the basis for national and international guidelines for the management of nearly all such patients in clinical practice. However, abundant and strict enrollment criteria may limit the validity and the applicability of results of randomized clinical trials to clinical practice. We estimated the eligibility for participation in landmark trials of antiplatelet drugs of an unselected group of patients with stroke or transient ischemic attack from a national stroke survey. METHODS: Nine hundred seventy-two patients with transient ischemic attack or ischemic stroke were prospectively and consecutively enrolled in the Netherlands Stroke Survey. We applied 7 large antiplatelet trials' enrollment criteria. RESULTS: In total, 886 patients were discharged alive and available for secondary prevention. Mean follow-up was 2.5 years. The annual rate of transient ischemic attack, stroke, or nonfatal myocardial infarction was 6.7%. The proportions of patients fulfilling the trial enrollment criteria ranged from 25% to 67%. Mortality was significantly higher in ineligible patients (27% to 41%) than in patients fulfilling enrollment criteria (16% to 20%). Rates of vascular events were not higher in trial-eligible patients than in ineligible patients. CONCLUSIONS: Our data confirm that patients with ischemic attack and stroke enrolled in randomized clinical trials are only partially representative of patients in clinical practice. Use of less strict enrollment criteria could enhance "generalizability" and result in more efficient selection of patients for randomized clinical trials. PMID- 19556534 TI - Outcome of symptomatic intracranial atherosclerotic disease. AB - BACKGROUND AND PURPOSE: Patients with intracranial atherosclerotic disease have a 3.6% to 22% annual risk of stroke. In this study, we sought to evaluate the natural history and prognosis of patients with symptomatic intracranial atherosclerotic disease who received medical therapy versus percutaneous transluminal angioplasty and stenting (PTAS) at our institution. METHODS: Charts of all patients with symptomatic intracranial atherosclerotic disease from July 2004 to September 2007 were reviewed and assessed for history of transient ischemic attack or stroke. Patients were either treated with "best medical therapy" (Medical Therapy Group) or PTAS plus antiplatelet agents (PTAS Group) and followed prospectively. A favorable outcome was defined as the absence of transient ischemic attacks, strokes, or vascular death; modified Rankin Scale of < or =3; and no endovascular reintervention of symptomatic in-stent restenosis. RESULTS: One hundred eleven patients fulfilled entry criteria, with 58 (52.3%) and 53 patients (47.7%) enrolled in the Medical Therapy and PTAS Groups, respectively. Thirty-eight patients of the Medical Therapy Group (65.5%) had a favorable outcome compared with 37 patients of the PTAS Group (69.8%). Combined ischemic end point data for the occurrence of transient ischemic attack, stroke, and vascular death was similar with 14 (24%) events in the Medical Therapy Group versus 15 (28.3%) events in the PTAS Group. CONCLUSIONS: Overall, the combined ischemic end point was the same in the Medical Therapy and PTAS Groups. PMID- 19556535 TI - Long-term functional recovery after first ischemic stroke: the Northern Manhattan Study. AB - BACKGROUND AND PURPOSE: Several factors predict functional status after stroke, but most studies have included hospitalized patients with limited follow-up. We hypothesized that patients with ischemic stroke experience functional decline over 5 years independent of recurrent stroke and other risk factors. METHODS: In the population-based Northern Manhattan Study, patients > or =40 years of age with incident ischemic stroke were prospectively followed using the Barthel Index at 6 months and annually to 5 years. Baseline stroke severity was categorized as mild (National Institutes of Health Stroke Scale <6), moderate (6 to 13), and severe (> or =14). Follow-up was censored at death, recurrent stroke, or myocardial infarction. Generalized Estimating Equations provided ORs and 95% CIs for predictors of favorable (Barthel Index > or =95) versus unfavorable (Barthel Index <95) functional status after adjusting for demographic and medical risk factors. RESULTS: Of 525 patients, mean age was 68.6+/-12.4 years, 45.5% were male, 54.7% Hispanic, 54.7% had Medicaid/no insurance, and 35.1% had moderate stroke. The proportion with Barthel Index > or =95 declined over time (OR, 0.91; 95% CI, 0.84 to 0.99). Changes in Barthel Index by insurance status were confirmed by a significant interaction term (beta for interaction=-0.167, P=0.034); those with Medicaid/no insurance declined (OR, 0.84; P=0.003), whereas those with Medicare/private insurance did not (OR, 0.99; P=0.92). CONCLUSIONS: The proportion of patients with functional independence after stroke declines annually for up to 5 years, and these effects are greatest for those with Medicaid or no health insurance. This decline is independent of age, stroke severity, and other predictors of functional decline and occurs even among those without recurrent stroke or myocardial infarction. PMID- 19556536 TI - The erythropoietin neuroprotective effect: assessment in CABG surgery (TENPEAKS): a randomized, double-blind, placebo controlled, proof-of-concept clinical trial. AB - BACKGROUND AND PURPOSE: Neurocognitive dysfunction complicates coronary artery bypass surgery. Erythropoietin may be neuroprotective. We sought to determine whether human recombinant erythropoietin would reduce the incidence of neurocognitive dysfunction after surgery. METHODS: We randomly assigned 32 elective first-time coronary artery bypass graft patients to receive placebo or 375 U/kg, 750 U/kg, or 1500 U/kg of recombinant human erythropoietin divided in 3 daily doses, starting the day before surgery. Primary outcomes were feasibility and safety, and secondary outcomes were neurocognitive dysfunction at discharge and 2 months. RESULTS: All subjects were male, mean age 60 years (range 46 to 73). No significant differences were found in pump time, cross-clamp time, or hospital length of stay. Mortality and pure red cell aplasia were not observed. One patient in the 375 U/kg group had ST changes compatible with myocardial injury immediately postoperative, but no other thrombotic complications were observed. Neurocognitive dysfunction occurred in 21/32 (66%) of patients at discharge and 5/32 (16%) at 2 months. Neurocognitive dysfunction at discharge by group was: placebo 6/8 (75%), 375 U/kg 4/8 (50%), 750 U/kg 6/8 (75%), and 1500 U/kg 5/8 (63%). Neurocognitive dysfunction at 2 months by group was: placebo 3/8 (38%), 375 U/kg 1/8 (13%), 750 U/kg 1/8 (13%), and 1500 U/kg 0/8 (0%). Neurocognitive dysfunction at 2 months for erythropoietin at any dose was 2/24 (8.3%) versus 3/8 (38%) for placebo (P=0.085). CONCLUSIONS: This study demonstrates feasibility and safety for the use of human recombinant erythropoietin as a neuroprotectant in coronary artery bypass graft surgery. A trend in the reduction of neurocognitive dysfunction at 2 months was associated with erythropoietin use. A multicenter randomized controlled trial is warranted. PMID- 19556537 TI - Impaired long-term stability of CA1 place cell representation in mice lacking the transcription factor zif268/egr1. AB - Zif268 is a transcriptional regulator that plays a crucial role in maintenance of the late phases of hippocampal long-term potentiation (LTP) and consolidation of spatial memories. Because the hippocampal place cell system is essential for long term spatial memory, we tested the hypothesis that zif268 is required for long term stability of hippocampal place cell representations by recording CA1 place cells in mice lacking zif268. We found that zif268 gene deletion destabilized the representation of a familiar environment after exposure to a novel environment and impaired the long-term (24 h), but not short-term (1 h), stability of newly formed representations. These impairments could be rescued by repeated exposure to the novel environment, however. These results indicate that zif268 contributes to the long-term stability of spatial representations in CA1 and support the notion that the long-term stability of place cell representations requires transcription-dependent mechanisms similar to those observed in LTP. PMID- 19556538 TI - Trim24 targets endogenous p53 for degradation. AB - Numerous studies focus on the tumor suppressor p53 as a protector of genomic stability, mediator of cell cycle arrest and apoptosis, and target of mutation in 50% of all human cancers. The vast majority of information on p53, its protein interaction partners and regulation, comes from studies of tumor-derived, cultured cells where p53 and its regulatory controls may be mutated or dysfunctional. To address regulation of endogenous p53 in normal cells, we created a mouse and stem cell model by knock-in (KI) of a tandem-affinity purification (TAP) epitope at the endogenous Trp-53 locus. Mass spectrometry of TAP-purified p53-complexes from embryonic stem cells revealed Tripartite-motif protein 24 (Trim24), a previously unknown partner of p53. Mutation of TRIM24 homolog, bonus, in Drosophila led to apoptosis, which could be rescued by p53 depletion. These in vivo analyses establish TRIM24/bonus as a pathway that negatively regulates p53 in Drosophila. The Trim24-p53 link is evolutionarily conserved, as TRIM24 depletion in human breast cancer cells caused p53-dependent, spontaneous apoptosis. We found that Trim24 ubiquitylates and negatively regulates p53 levels, suggesting Trim24 as a therapeutic target to restore tumor suppression by p53. PMID- 19556539 TI - Extinction implications of a chenopod browse diet for a giant Pleistocene kangaroo. AB - Kangaroos are the world's most diverse group of herbivorous marsupials. Following late-Miocene intensification of aridity and seasonality, they radiated across Australia, becoming the continent's ecological equivalents of the artiodactyl ungulates elsewhere. Their diversity peaked during the Pleistocene, but by approximately 45,000 years ago, 90% of larger kangaroos were extinct, along with a range of other giant species. Resolving whether climate change or human arrival was the principal extinction cause remains highly contentious. Here we combine craniodental morphology, stable-isotopic, and dental microwear data to reveal that the largest-ever kangaroo, Procoptodon goliah, was a chenopod browse specialist, which may have had a preference for Atriplex (saltbushes), one of a few dicots using the C(4) photosynthetic pathway. Furthermore, oxygen isotope signatures of P. goliah tooth enamel show that it drank more in low-rainfall areas than its grazing contemporaries, similar to modern saltbush feeders. Saltbushes and chenopod shrublands in general are poorly flammable, so landscape burning by humans is unlikely to have caused a reduction in fodder driving the species to extinction. Aridity is discounted as a primary cause because P. goliah evolved in response to increased aridity and disappeared during an interval wetter than many it survived earlier. Hunting by humans, who were also bound to water, may have been a more decisive factor in the extinction of this giant marsupial. PMID- 19556540 TI - Hypertension of Kcnmb1-/- is linked to deficient K secretion and aldosteronism. AB - Mice lacking the beta1-subunit (gene, Kcnmb1; protein, BK-beta1) of the large Ca activated K channel (BK) are hypertensive. This phenotype is thought to result from diminished BK currents in vascular smooth muscle where BK-beta1 is an ancillary subunit. However, the beta1-subunit is also expressed in the renal connecting tubule (CNT), a segment of the aldosterone-sensitive distal nephron, where it associates with BK and facilitates K secretion. Because of the correlation between certain forms of hypertension and renal defects, particularly in the distal nephron, it was determined whether the hypertension of Kcnmb1(-/-) has a renal origin. We found that Kcnmb1(-/-) are hypertensive, volume expanded, and have reduced urinary K and Na clearances. These conditions are exacerbated when the animals are fed a high K diet (5% K; HK). Supplementing HK-fed Kcnmb1(-/ ) with eplerenone (mineralocorticoid receptor antagonist) corrected the fluid imbalance and more than 70% of the hypertension. Finally, plasma [aldo] was elevated in Kcnmb1(-/-) under basal conditions (control diet, 0.6% K) and increased significantly more than wild type when fed the HK diet. We conclude that the majority of the hypertension of Kcnmb1(-/-) is due to aldosteronism, resulting from renal potassium retention and hyperkalemia. PMID- 19556542 TI - Statistical approach to quantifying the elastic deformation of nanomaterials. AB - Quantifying the mechanical properties of nanomaterials is challenged by its small size, difficulty of manipulation, lack of reliable measurement techniques, and grossly varying measurement conditions and environment. A recently proposed approach is to estimate the elastic modulus from a force-deflection physical model based on the continuous bridged-deformation of a nanobelt/nanowire using an atomic force microscope tip under different contact forces. However, the nanobelt may have some initial bending, surface roughness and imperfect physical boundary conditions during measurement, leading to large systematic errors and uncertainty in data quantification. In this article, a statistical modeling technique, sequential profile adjustment by regression (SPAR), is proposed to account for and eliminate the various experimental errors and artifacts. SPAR can automatically detect and remove the systematic errors and therefore gives more precise estimation of the elastic modulus. This research presents an innovative approach that can potentially have a broad impact in quantitative nanomechanics and nanoelectronics. PMID- 19556541 TI - Structural and molecular basis of the assembly of the TRPP2/PKD1 complex. AB - Mutations in PKD1 and TRPP2 account for nearly all cases of autosomal dominant polycystic kidney disease (ADPKD). These 2 proteins form a receptor/ion channel complex on the cell surface. Using a combination of biochemistry, crystallography, and a single-molecule method to determine the subunit composition of proteins in the plasma membrane of live cells, we find that this complex contains 3 TRPP2 and 1 PKD1. A newly identified coiled-coil domain in the C terminus of TRPP2 is critical for the formation of this complex. This coiled coil domain forms a homotrimer, in both solution and crystal structure, and binds to a single coiled-coil domain in the C terminus of PKD1. Mutations that disrupt the TRPP2 coiled-coil domain trimer abolish the assembly of both the full-length TRPP2 trimer and the TRPP2/PKD1 complex and diminish the surface expression of both proteins. These results have significant implications for the assembly, regulation, and function of the TRPP2/PKD1 complex and the pathogenic mechanism of some ADPKD-producing mutations. PMID- 19556544 TI - Molecular noise of capping protein binding induces macroscopic instability in filopodial dynamics. AB - Capping proteins are among the most important regulatory proteins involved in controlling complicated stochastic dynamics of filopodia, which are dynamic finger-like protrusions used by eukaryotic motile cells to probe their environment and help guide cell motility. They attach to the barbed end of a filament and prevent polymerization, leading to effective filament retraction due to retrograde flow. When we simulated filopodial growth in the presence of capping proteins, qualitatively different dynamics emerged, compared with actin only system. We discovered that molecular noise due to capping protein binding and unbinding leads to macroscopic filopodial length fluctuations, compared with minuscule fluctuations in the actin-only system. Thus, our work shows that molecular noise of signaling proteins may induce micrometer-scale growth retraction cycles in filopodia. When capped, some filaments eventually retract all the way down to the filopodial base and disappear. This process endows filopodium with a finite lifetime. Additionally, the filopodia transiently grow several times longer than in actin-only system, since less actin transport is required because of bundle thinning. We have also developed an accurate mean field model that provides qualitative explanations of our numerical simulation results. Our results are broadly consistent with experiments, in terms of predicting filopodial growth retraction cycles and the average filopodial lifetimes. PMID- 19556546 TI - ESR evidence for 2 coexisting liquid phases in deeply supercooled bulk water. AB - Using electron spin resonance spectroscopy (ESR), we measure the rotational mobility of probe molecules highly diluted in deeply supercooled bulk water and negligibly constrained by the possible ice fraction. The mobility increases above the putative glass transition temperature of water, T(g) = 136 K, and smoothly connects to the thermodynamically stable region by traversing the so called "no man's land" (the range 150-235 K), where it is believed that the homogeneous nucleation of ice suppresses the liquid water. Two coexisting fractions of the probe molecules are evidenced. The 2 fractions exhibit different mobility and fragility; the slower one is thermally activated (low fragility) and is larger at low temperatures below a fragile-to-strong dynamic cross-over at approximately 225 K. The reorientation of the probe molecules decouples from the viscosity below approximately 225 K. The translational diffusion of water exhibits a corresponding decoupling at the same temperature [Chen S-H, et al. (2006) The violation of the Stokes-Einstein relation in supercooled water. Proc Natl Acad Sci USA 103:12974-12978]. The present findings are consistent with key issues concerning both the statics and the dynamics of supercooled water, namely the large structural fluctuations [Poole PH, Sciortino F, Essmann U, Stanley HE (1992) Phase behavior of metastable water. Nature 360:324-328] and the fragile-to strong dynamic cross-over at approximately 228 K [Ito K, Moynihan CT, Angell CA (1999) Thermodynamic determination of fragility in liquids and a fragile-to strong liquid transition in water. Nature 398:492-494]. PMID- 19556545 TI - DNA methylation is widespread and associated with differential gene expression in castes of the honeybee, Apis mellifera. AB - The recent, unexpected discovery of a functional DNA methylation system in the genome of the social bee Apis mellifera underscores the potential importance of DNA methylation in invertebrates. The extent of genomic DNA methylation and its role in A. mellifera remain unknown, however. Here we show that genes in A. mellifera can be divided into 2 distinct classes, one with low-CpG dinucleotide content and the other with high-CpG dinucleotide content. This dichotomy is explained by the gradual depletion of CpG dinucleotides, a well-known consequence of DNA methylation. The loss of CpG dinucleotides associated with DNA methylation also may explain the unusual mutational patterns seen in A. mellifera that lead to AT-rich regions of the genome. A detailed investigation of this dichotomy implicates DNA methylation in A. mellifera development. High-CpG genes, which are predicted to be hypomethylated in germlines, are enriched with functions associated with developmental processes, whereas low-CpG genes, predicted to be hypermethylated in germlines, are enriched with functions associated with basic biological processes. Furthermore, genes more highly expressed in one caste than another are overrepresented among high-CpG genes. Our results highlight the potential significance of epigenetic modifications, such as DNA methylation, in developmental processes in social insects. In particular, the pervasiveness of DNA methylation in the genome of A. mellifera provides fertile ground for future studies of phenotypic plasticity and genomic imprinting. PMID- 19556547 TI - eIF4A controls germline stem cell self-renewal by directly inhibiting BAM function in the Drosophila ovary. AB - Stem cell self-renewal is controlled by concerted actions of extrinsic niche signals and intrinsic factors in a variety of systems. Drosophila ovarian germline stem cells (GSCs) have been one of the most productive systems for identifying the factors controlling self-renewal. The differentiation factor BAM is necessary and sufficient for GSC differentiation, but it still remains expressed in GSCs at low levels. However, it is unclear how its function is repressed in GSCs to maintain self-renewal. Here, we report the identification of the translation initiation factor eIF4A for its essential role in self-renewal by directly inactivating BAM function. eIF4A can physically interact with BAM in Drosophila S2 cells and yeast cells. eIF4A exhibits dosage-specific interactions with bam in the regulation of GSC differentiation. It is required intrinsically for controlling GSC self-renewal and proliferation but not survival. In addition, it is required for maintaining E-cadherin expression but not BMP signaling activity. Furthermore, BAM and BGCN together repress translation of E-cadherin through its 3' UTR in S2 cells. Therefore, we propose that BAM functions as a translation repressor by interfering with translation initiation and eIF4A maintains self-renewal by inhibiting BAM function and promoting E-cadherin expression. PMID- 19556548 TI - Independent component analysis for brain fMRI does not select for independence. AB - InfoMax and FastICA are the independent component analysis algorithms most used and apparently most effective for brain fMRI. We show that this is linked to their ability to handle effectively sparse components rather than independent components as such. The mathematical design of better analysis tools for brain fMRI should thus emphasize other mathematical characteristics than independence. PMID- 19556549 TI - Metal-independent decomposition of hydroperoxides by halogenated quinones: detection and identification of a quinone ketoxy radical. AB - We have shown recently that halogenated quinones could enhance the decomposition of hydroperoxides and formation of alkoxyl/hydroxyl radicals through a metal independent mechanism. However, neither the proposed quinone enoxy radical intermediate, nor the major reaction products were unambiguously identified. In the present study, one of the major reaction products between 2,5-dichloro-1,4 benzoquinone (DCBQ) and t-butylhydroperoxide (t-BuOOH) was isolated and purified by semipreparative HPLC, and identified as 2-hydroxy-3-t-butoxy-5-chloro-1,4 benzoquinone [CBQ(OH)-O-t-Bu], which is the rearranged isomer of the postulated quinone-peroxide reaction intermediate. The formation of CBQ(OH)-O-t-Bu was found to be inhibited by the spin trapping agent 5,5-dimethyl-1-pyrroline N-oxide (DMPO), and concurrently, a new DMPO adduct with 1-chlorine isotope peak clusters at m/z 268 was observed. Further electron spin resonance (ESR) spin-trapping, (1)H-NMR and HPLC/Fourier transform ion cyclotron resonance (FTICR) mass spectrometric studies with oxygen-17-labeled and unlabeled hydrogen peroxide strongly suggest that the radical trapped by DMPO is a carbon-centered quinone ketoxy radical, which is the spin isomer of the proposed oxygen-centered quinone enoxy radical. Analogous results were observed when DCBQ was substituted by other halogenated quinones. This study represents the first detection and identification of an unusual carbon-centered quinone ketoxy radical, which provides direct experimental evidence to further support and expand our previously proposed mechanism for metal-independent decomposition of hydroperoxides by halogenated quinones. PMID- 19556550 TI - High cGMP synthetic activity in carp cones. AB - Cones show briefer light responses than rods and do not saturate even under very bright light. Using purified rod and cone homogenates, we measured the activity of guanylate cyclase (GC), an enzyme responsible for cGMP synthesis and therefore recovery of a light response. The basal GC activity was 36 times higher in cones than in rods: It was mainly caused by higher expression levels of GC in cones (GC C) than in rods (GC-R). With identification and quantification of GC-activating protein (GCAP) subtypes expressed in rods and cones together with determination of kinetic parameters of GC activation in the presence and absence of GCAP, we estimated the in situ GC activity in rods and cones at low and high Ca(2+) concentrations. It was revealed that the GC activity would be >10 times higher in cones than in rods in both the dark-adapted and the light-adapted states. Electrophysiological estimation of the GC activity measured in the truncated preparations of rod and cone outer segments gave consistent results. Our estimation of the in situ GC activity reasonably explained the rapid recovery and nonsaturating behavior of cone light responses. PMID- 19556551 TI - The macroscopic delamination of thin films from elastic substrates. AB - The wrinkling and delamination of stiff thin films adhered to a polymer substrate have important applications in "flexible electronics." The resulting periodic structures, when used for circuitry, have remarkable mechanical properties because stretching or twisting of the substrate is mostly accommodated through bending of the film, which minimizes fatigue or fracture. To date, applications in this context have used substrate patterning to create an anisotropic substrate film adhesion energy, thereby producing a controlled array of delamination "blisters." However, even in the absence of such patterning, blisters appear spontaneously, with a characteristic size. Here, we perform well-controlled experiments at macroscopic scales to study what sets the dimensions of these blisters in terms of the material properties and explain our results by using a combination of scaling and analytical methods. Besides pointing to a method for determining the interfacial toughness, our analysis suggests a number of design guidelines for the thin films used in flexible electronic applications. Crucially, we show that, to avoid the possibility that delamination may cause fatigue damage, the thin film thickness must be greater than a critical value, which we determine. PMID- 19556552 TI - Perception of barriers to self-care management among diabetic patients. AB - PURPOSE: The purpose of this study was to explore individual, educational, and system barriers that limit low-income diabetes patients' ability to achieve optimal diabetes self-management. METHODS: Economically disadvantaged patients with diabetes who used the Diabetes Clinic of Grady Health System in Atlanta, Georgia, participated in 3 focus group discussions. RESULTS: The discussions were held with mostly African Americans (n = 35) to explore barriers to achieving optimal diabetes self-management. Most participants were not married, approximately one-third had less than high school level reading skills, and 40% were not currently working. In terms of individual barriers, the emotional toll from the diagnosis of and lifestyle changes to treat diabetes was a recurrent theme, and included stress, frustration, social isolation, interpersonal conflicts, depression, and fear. Denial was often mentioned as the key factor that inhibited adherence to a healthy mode of living. The educational barriers were failure to recognize the risks and consequences of an asymptomatic condition. Many participants did not understand A1C. Finally, several system barriers were identified. The participants identified needed services, including follow-up and refresher courses, support group discussions, nutrition and medication education, availability of different education modalities, and expanded clinic hours. CONCLUSIONS: The focus group discussions identified both barriers to diabetes management and opportunities for improving care for underserved patients with diabetes. The results are useful to improve the delivery of care and to develop quantitative studies to explore particular areas of interest. Based on these results, the current system needs to provide more support and education to patients with diabetes. PMID- 19556553 TI - Understanding observed and unobserved health care access and utilization disparities among US Latino adults. AB - This study hypothesizes that differences in health care access and utilization exist across Latino adults (>18 years), with U.S. Latino adults of Mexican ancestry demonstrating the worst patterns of access and utilization. The analyses use the National Health Interview Survey (NHIS) data from 1999 to 2007 (N = 33,908). The authors first estimate the disparities in health care access and utilization among different categories of Latinos. They also implement Blinder Oaxaca techniques to decompose disparities into observed and unobserved components, comparing Latinos of Mexican ancestry with non-Mexican Latinos. Latinos of Mexican ancestry consistently demonstrate lower health care access and utilization patterns than non-Mexican Latinos. Health insurance and region of residence were the most important factors that explained observable differences. In contrast, language and citizenship status were relatively unimportant. Although a significant share of these disparities may be explained by observed characteristics, disparities because of unobserved heterogeneity among the different Latino cohorts are also considerable. PMID- 19556554 TI - Determinants of hospitalist efficiency: a qualitative and quantitative study. AB - Using qualitative and quantitative methods, the authors develop and test hypotheses about the impact of hospitalists on efficiency and quality of care relative to teaching teams. Departure of actual from self-perceived benefits for hospitalists, both individually and collectively, is studied. It was found that hospitalists are, on average, more efficient diagnosticians and/or enhance throughput, as evidenced by having relatively lower charges, through reductions in testing and length-of-stay, than teaching teams. Much of that benefit is concentrated among patients admitted by intensivists. The authors find little evidence of quality focus or of greater use of community resources among hospitalists. Indeed, hospitalists were found to have no effect on the choice of postdischarge outlets. The authors document variation in care delivery among hospitalists. In particular, it was found that among hospitalists there is more variation in achieving shorter length of stay but less variation in use of diagnostic testing. PMID- 19556555 TI - Studying the interpretation of dreams in the company of analytic candidates. AB - Seminars serve as an important, though undervalued, component of psychoanalytic education. The focus of this paper is on the teaching of Freud's The Interpretation of Dreams through a series of seminars presented to analytic candidates at the Toronto Psychoanalytic Institutes. This has been an essential book for introducing generations of candidates to the psychoanalytic concept of the mind and for shaping candidates' understanding and attitudes toward working with their patients' dreams. Four of Freud's basic dream concepts-(1) the method and its application to the exploration of the relationship between manifest and latent dream content, (2) the sources of dreams (day residues), (3) the dream work, and (4) wish fulfillment-are critically studied in the seminars. Detailed discussion of these basic dream concepts among the candidates and with the teacher, as well as the candidates' feedback at the conclusion of the seminars, are summarized and discussed. Through the teaching and study within the seminar framework of the fundamentals of Freud's dream theory, a shared growth experience results for both teacher and candidates. PMID- 19556557 TI - A case of optic nerve compression caused by Angiostrongylus cantonensis. PMID- 19556558 TI - Linking deforestation to malaria in the Amazon: characterization of the breeding habitat of the principal malaria vector, Anopheles darlingi. AB - This study examined the larval breeding habitat of a major South American malaria vector, Anopheles darlingi, in areas with varying degrees of ecologic alteration in the Peruvian Amazon. Water bodies were repeatedly sampled across 112 km of transects along the Iquitos-Nauta road in ecologically varied areas. Field data and satellite imagery were used to determine the landscape composition surrounding each site. Seventeen species of Anopheles larvae were collected. Anopheles darlingi larvae were present in 87 of 844 sites (10.3%). Sites with A. darlingi larvae had an average of 24.1% forest cover, compared with 41.0% for sites without A. darlingi (P < 0.0001). Multivariate analysis identified seasonality, algae, water body size, presence of human populations, and the amount of forest and secondary growth as significant determinants of A. darlingi presence. We conclude that deforestation and associated ecologic alterations are conducive to A. darlingi larval presence, and thereby increase malaria risk. PMID- 19556559 TI - Molecular epidemiology of malaria in Cameroon. XXVIII. In vitro activity of dihydroartemisinin against clinical isolates of Plasmodium falciparum and sequence analysis of the P. falciparum ATPase 6 gene. AB - The Plasmodium falciparum ATPase 6 (Pfatp6), homolog of sarco-endoplasmic reticulum, calcium-dependent ATPase in malaria parasites, has been proposed to be the main target of artemisinins. Four distinct point mutations (L263E, E431K, A623E, and S769N) have been reported to be associated with artemisinin resistance. The Pfatp6 sequence polymorphism was determined to evaluate the prevalence of these mutations in fresh clinical isolates in Yaounde, Cameroon, and compare sequence data with in vitro response to dihydroartemisinin. Two major haplotypes were observed: the wild-type LEAS (n = 60, 62%) and a single mutant LKAS (n = 35, 36%). These amino acid substitutions did not influence the level of in vitro response to dihydroartemisinin (P > 0.05). Plasmodium falciparum isolates from Cameroon are highly sensitive in vitro to artemisinins. However, the relatively high prevalence of E431K may be a warning signal that warrants a regular monitoring of these molecular markers and/or in vitro activity of artemisinin derivatives. PMID- 19556560 TI - High prevalence and fixation of Plasmodium vivax dhfr/dhps mutations related to sulfadoxine/pyrimethamine resistance in French Guiana. AB - Plasmodium vivax isolates from French Guiana were studied for the presence of mutations associated with sulfadoxine/pyrimethamine (SP) drug resistance. Ninety six blood samples were collected from 2000 to 2005 from symptomatic malaria patients. SP drug resistance was predicted by determining point mutations in the dihydrofolate reductase (pvdhfr) and dihydropteroate synthase (pvdhps) genes. All samples showed mutant genotypes in both genes with a prevalence > 90% for the 58R, 117N, 382C, and 383G. A new mutation (116G) in pvdhfr was found at a frequency of 3.3%. Six different pvdhfr/dhps multilocus genotypes were observed with the predominance of the quintuple mutant-type 58R/117N/173L-382C/383G (59.3%). No significant differences were observed between the prevalence of haplotypes and the year of collection. Our results indicate that, in this area, the fixation of SP drug-resistant parasites in the P. vivax population is stable. PMID- 19556561 TI - Anopheles darlingi (Diptera: Culicidae) in Panama. AB - We report Anopheles darlingi in Darien Province in eastern Panama. Polymerase chain reaction-restriction fragment length polymorphism profiles of the single copy nuclear white gene and sequence comparisons confirmed the presence of 66 specimens of the northern lineage of An. darlingi. The parsimony network depicted 5 CO1 haplotypes in 40 specimens of An. darlingi, which connected through 7-8 mutational steps with sequences from Central and South America. Furthermore, the presence of haplotypes in Biroquera, Darien Province identical to those previously published from northern Colombia suggests that Panamanian samples originated in Colombia. Results of neutrality tests (R(2) and Fu's F(S)) were not significant and the mismatch distribution was multimodal and did not fit the model of sudden population growth. These findings may indicate a long and stable presence of An. darlingi in eastern Panama. PMID- 19556562 TI - Comparative study of serologic tests for the diagnosis of asymptomatic visceral leishmaniasis in an endemic area. AB - Serologic tests have been widely used for the diagnosis of asymptomatic visceral leishmaniasis. This study evaluated five serologic tests used for the diagnosis of asymptomatic infection: enzyme-linked immunosorbent assay (ELISA) using promastigote antigen (ELISAp), ELISA using recombinant K39 (ELISA rK39), and K26 (ELISA rK26) antigens, an indirect immunofluorescence test using Leishmania (Leishmania) amazonensis promastigote antigen (IIFT), and an immunochromatographic test using rK39 antigen (TRALd). As a reference regarding the performance of the tests, patients with classic visceral leishmaniasis originating from Minas Gerais, Brazil (N = 36), were defined as the positive group and samples of healthy individuals from nonendemic areas (Argentina) (N = 127) were used as negative controls. Patients with other diseases such as cutaneous leishmaniasis (N = 53) and malaria (N = 56) were also studied to evaluate the chance of cross-reactivity in these tests. Finally, subjects from an area endemic for visceral leishmaniasis in Brazil (Porteirinha, northern Minas Gerais) (N = 1241) were screened for asymptomatic infection with Leishmania and Chagas disease. The sensitivity of the serologic tests was 50% (18/36), 66.7% (24/36), 69.4% (25/36), 83.3% (30/36), and 88.9% (32/36) for ELISAp, ELISA rK26, ELISA rK39, IIFT, and TRALd, respectively. Specificity, calculated using the truly negative group, was 96% (122/127) for TRALd, 97.6% (124/127) for ELISAp and IIFT, and 100% (127/127) for ELISA rK39 and rK26. Positivity in at least one test employing recombinant antigen was observed in 24 (45%) patients with cutaneous leishmaniasis and 47 (82.4%) with malaria. In the visceral leishmaniasis-endemic area, the positivity of the serologic tests ranged from 3.9% to 37.5%. The enzyme linked immunosorbent assay (ELISA) tests using recombinant antigens were more frequently positive in subjects with a history of exposure to human or canine visceral leishmaniasis (ELISArK39: 14.6% [149/1017] versus 37.5% [84/224]; ELISA rK26: 12.7% [129/1017] versus 21.4% [48/224], P < 0.001 for both). Kappa agreement was low, with a maximum value of 0.449 between ELISAp and IIFT. In addition, among the 112 IIFT-positive subjects, 75 (67%) also presented positive serology for Chagas disease. In conclusion, IIFT and TRALd presented the best performance to diagnose classic cases of visceral leishmaniasis in an endemic area. Cross-reactivity of the tests with Chagas disease, cutaneous leishmaniasis, and malaria should be taken into account. However, the differences in the positivity of the tests used, together with the low agreement between results, do not permit to select the best test for the diagnosis of asymptomatic Leishmania infection. PMID- 19556564 TI - Visceral leishmaniasis in Tunisia: spatial distribution and association with climatic factors. AB - Visceral leishmaniasis (VL) cases in children less than five years of age were recorded from 1996 through 2006 from Tunisian pediatric departments. Mean incidence rates were calculated for each of the 215 districts in the study area. Averages of annual rainfall and extreme values of low temperatures in winter and high temperatures in summer were used to characterize the climate of each district according to its continentality index and bioclimatic zone. A geographic information system and a local indicator of spatial association were used to summarize the spatial properties of VL distribution. Poisson spatial regression was performed to study the relationship between VL incidence rates and climatic parameters. We identified one hot-spot region of 35 inland districts located mostly in the semi-arid bioclimatic zone and two cold-spots located in coastal regions of the northeastern sub-humid zone and the southeastern arid zone. The incidence rate of VL was positively correlated with mean yearly rainfall and continentality index. PMID- 19556563 TI - Risk factors for visceral leishmaniasis in a new epidemic site in Amhara Region, Ethiopia. AB - We conducted a case-control study to evaluate risk factors for visceral leishmaniasis during an epidemic in a previously unaffected district of Ethiopia. We also collected blood and bone marrow specimens from dogs in the outbreak villages. In multivariable analyses of 171 matched case-control pairs, dog ownership, sleeping under an acacia tree during the day, and habitually sleeping outside at night were associated with significantly increased risk. Specimens from 7 (3.8%) dogs were positive by immunofluorescent antibody test (IFAT) and both enzyme-linked immunosorbent assays (ELISAs), whereas Leishmania DNA was detected in 5 (2.8%) bone marrow aspirates (from 3 seropositive and 2 seronegative dogs). Insecticide-treated nets may only protect a portion of those at risk. Further research on the vectors, the role of the dog in the transmission cycle, and the effect of candidate interventions are needed to design the best strategy for control. PMID- 19556565 TI - Is real-time polymerase chain reaction (PCR) more useful than a conventional PCR for the clinical management of leishmaniasis? AB - It is currently unknown if the use of a real-time polymerase chain reaction (PCR) adds value to the diagnosis and follow-up prognosis of patients affected by leishmaniasis. We performed a study using a real-time PCR directed against the alpha-polymerase gene and a semiquantitative PCR that target the SSU ribosomal RNA (rRNA) gene as control for the diagnosis and quantification of parasites in patients with visceral (VL) and cutaneous (CL) leishmaniasis. Our single copy real-time PCR missed one diagnosis of VL compared with the conventional PCR, whereas both PCR methods were able to detect Leishmania parasites in CL. Under anti-leishmania treatment the kinetics of parasitemia were comparable with the two methods. The real-time PCR directed against alpha-polymerase of Leishmania despite being able to make a more accurate quantification of parasites does not add to the decision-making management compared with a semiquantitative PCR, and it is comparatively expensive. PMID- 19556566 TI - Relapsing cutaneous leishmaniasis in a patient with ankylosing spondylitis treated with infliximab. AB - A 31-year-old man with ankylosing spondylitis, receiving treatment with infliximab, presented with a large progressive cutaneous ulcer at the right knee. Biopsies showed Leishmania amastigotes, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis showed Leishmania infantum as the causative agent. After treatment with miltefosine, the ulcer resolved completely, and infliximab was reinstituted because of progression of spondylitis. After 1 year, there was a recurrent ulcer at the same site being positive for Leishmania DNA by PCR. Local treatment with sodium stibogluconate resulted in complete regression. Cutaneous leishmaniasis should be added to the list of opportunistic infections associated with anti-tumor necrosis factor (TNF) treatment. Despite recurrences, antileish-manial treatment may be effective in cases without alternatives to anti-TNF therapy. PMID- 19556567 TI - Isolation and genetic characterization of a Bartonella strain closely related to Bartonella tribocorum and Bartonella elizabethae in Israeli commensal rats. AB - Ten Bartonella isolates were cultured from blood drawn from black rats (Rattus rattus) captured in the Tel Aviv area. Genetic characterization included amplification and sequencing of five gene fragments including the ribC, rpoB, 16S, groEL, and gltA and the 16S-23S intergenic spacer region. Sequence comparisons showed that all 10 isolates were identical in all genes studied comprising a total of 3,873 bp analyzed. The sequences of each of the partial genes analyzed indicated a high sequence similarity (97-99.8%) to B. tribocorum or B. elizabethae. The gltA sequence was 100% homologous to a genotype identified in R. rattus in Dhaka, Bangladesh, suggesting the existence of a widespread Asian Bartonella strain infecting the black rats (R. rattus). The detection of a Bartonella genotype closely related to B. elizabethae in the biggest metropolitan center in Israel warrants further study of its zoonotic potential and pathogenic characteristics. PMID- 19556569 TI - Tick-borne zoonotic bacteria in ticks collected from central Spain. AB - The prevalence of tick-borne and related bacteria infecting adult ticks in central Spain was assessed by molecular methods. Six areas were sampled monthly during a 2-year longitudinal study. A total of 1,038 questing and 442 feeding ticks, belonging to eight different species, were tested. The most abundant species were Hyalomma lusitanicum (54% of captures), followed by Dermacentor marginatus (23%) and Rhipicephalus sanguineus (10%). Four human pathogens, including seven Rickettsia species, Anaplasma phagocytophilum, Borrelia burgdorferi, and Francisella tularensis, were detected at percentages of 19.0, 2.2, 1.7, and 0.5, respectively, whereas Bartonella spp. was never detected. In terms of infection and tick abundance, H. lusitanicum seems to be the most significant tick species in the area, carrying three of the five agents tested, and the anthropophilic tick, D. marginatum, infected with Rickettsia spp. and F. tularensis, is the most relevant in terms of public health. The significance of these data is discussed. PMID- 19556568 TI - Hantavirus infection and habitat associations among rodent populations in agroecosystems of Panama: implications for human disease risk. AB - Hantavirus cardiopulmonary syndrome (HCPS), which is caused by infection with Choclo virus, is uncommon in Panama, yet seropositivity among rural residents is as high as 60%. To clarify the environmental risk factors favoring rodent-to human transmission, we tested serum from 3,067 rodents captured over a five-year period for antibodies against recombinant N protein of hantavirus by enzyme immunoassay and strip immunoblot. Among 220 seropositive rodents, Oligoryzomys fulvescens, the reservoir of Choclo virus, had the highest overall seroprevalence (23.5%); more abundant rodents (Zygodontomys brevicauda and Sigmodon hirsutus) had lower seroprevalences. In the mixed (combined modern and traditional) productive agroecosystem, the highest seroprevalence was among O. fulvescens captured in residences and in crops grown within 40 meters of a residence, with significantly lower seroprevalence in adjacent pasture and non-productive vegetation. Thus, crop habitats may serve as refugia for invasion into adjacent human residences and suggests several interventions to reduce human infection. PMID- 19556570 TI - Outcome of patients with buruli ulcer after surgical treatment with or without antimycobacterial treatment in Ghana. AB - This study assesses the frequency of recurrences and treatment outcome after surgery of buruli ulcer disease (BUD) with or without concomitant antimycobacterial treatment. Of 129 laboratory-confirmed BUD patients who underwent surgery in two treatment centers in Ghana, 79 (61%) were retrieved for follow-up 4-29 months after the initial treatment. Among 7 (9%) recurrent cases no significant association was found between recurrences and clinical or treatment specific factors including antimycobacterial treatment. In 21 (27%) patients, a reduced range of motion (ROM) of one or more joints was detected. Lesions other than nodules, joint involvement, and skin grafting were identified as independent risk factors. Functional limitations hampering daily activities were perceived by 22% of the patients. Compared with other studies the recurrence rate was relatively low, functional limitations were, however, frequent. This emphasizes the need for improvement of pre- and post-treatment wound care as well as rehabilitation programs. PMID- 19556571 TI - Functional limitations after surgical or antibiotic treatment for Buruli ulcer in Benin. AB - Almost half of patients have functional limitations after treatment of Buruli ulcer disease. Antibiotic treatment (along with surgery) was introduced in the National Program for Buruli ulcer in Benin in 2005. The aim of this study was to compare functional limitations in patients who were treated by antibiotics, surgery, or both, using a validated questionnaire. One hundred seventy-nine former patients in Lalo, Benin were retrieved and interviewed in their village. Hospital records were used to gather data about size of lesion at presentation and treatment provided. No significant differences in resulting functional limitations were found between the different treatments. Larger lesions (> 15 cm cross-sectional diameter) at presentation; lesions on a joint, muscular atrophy, and amputation were all associated with a higher risk for functional limitations. Advantages of antibiotic treatment may involve other domains, like costs of treatment or a change in help-seeking behavior. PMID- 19556572 TI - Cost-effectiveness of a community-based approach intertwined with a vertical Aedes control program. AB - We compared in a 5-year intervention study the cost-effectiveness of community based environmental management intertwined with routine vertical Aedes control and of routine vertical control only. At baseline (year 2000), Aedes infestation levels and economic costs for vector control were comparable in intervention and control areas (house index, 2.23% versus 2.21% and US$21 versus US$24/yr/inhabitant, respectively). By 2004, house indices became 0.22% versus 2.36% and the costs were 29.8 US$ versus 36.7 US$/yr/inhabitant, respectively. The community cost made up 38.6% of the total economic cost in 2004 in the intervention areas against 23.5% in 2000. The average cost-effectiveness ratio for the intervention period 2001-2004, expressed as the societal cost incurred for the reduction (from baseline) of Aedes foci, was US$831.1 per focus in the intervention areas versus US$2,465.6 in the control areas. The intervention produced economic savings and health benefits that were sustained over the whole observation period. PMID- 19556573 TI - The effects of sustained release metofluthrin on the biting, movement, and mortality of Aedes aegypti in a domestic setting. AB - The impact of a sustained release metofluthrin emanator and an allethrin-based mosquito coil on biting, movement and mortality of female Aedes aegypti was assessed in an apartment. In the room in which the metofluthrin emanator was activated, mosquito biting counts were reduced to zero. Metofluthrin also had a spillover effect, significantly (P < 0.001) reducing biting counts in a neighboring room 1, 4, and 24 hours after the emanator was activated when compared with either the coil or control (untreated) treatment. Mosquitoes were neither repelled nor expelled from a room exposed to metofluthrin. Indeed, a significantly (P = 0.023) greater proportion of mosquitoes were found in the treated room after exposure to metofluthrin when compared with either the coil or control treatment. Furthermore, in the room treated with metofluthrin the majority of mosquitoes died and a spillover effect into the neighboring room caused greater than one-third mortality of the mosquitoes. Metofluthrin could be used to prevent dengue transmission within a household. PMID- 19556574 TI - Simplified pupal surveys of Aedes aegypti (L.) for entomologic surveillance and dengue control. AB - Pupal surveys of Aedes aegypti (L.) are useful indicators of risk for dengue transmission, although sample sizes for reliable estimations can be large. This study explores two methods for making pupal surveys more practical yet reliable and used data from 10 pupal surveys conducted in Puerto Rico during 2004-2008. The number of pupae per person for each sampling followed a negative binomial distribution, thus showing aggregation. One method found a common aggregation parameter (k) for the negative binomial distribution, a finding that enabled the application of a sequential sampling method requiring few samples to determine whether the number of pupae/person was above a vector density threshold for dengue transmission. A second approach used the finding that the mean number of pupae/person is correlated with the proportion of pupa-infested households and calculated equivalent threshold proportions of pupa-positive households. A sequential sampling program was also developed for this method to determine whether observed proportions of infested households were above threshold levels. These methods can be used to validate entomological thresholds for dengue transmission. PMID- 19556575 TI - Voltage-gated sodium channel polymorphism and metabolic resistance in pyrethroid resistant Aedes aegypti from Brazil. AB - The nature of pyrethroid resistance in Aedes aegypti Brazilian populations was investigated. Quantification of enzymes related to metabolic resistance in two distinct populations, located in the Northeast and Southeast regions, revealed increases in Glutathione-S-transferase (GST) and Esterase levels. Additionally, polymorphism was found in the IIS6 region of Ae. aegypti voltage-gated sodium channel (AaNa(V)), the pyrethroid target site. Sequences were classified in two haplotype groups, A and B, according to the size of the intron in that region. Rockefeller, a susceptible control lineage, contains only B sequences. In field populations, some A sequences present a substitution in the 1011 site (Ile/Met). When resistant and susceptible individuals were compared, the frequency of both A (with the Met mutation) and B sequences were slightly increased in resistant specimens. The involvement of the AaNa(V) polymorphism in pyrethroid resistance and the metabolic mechanisms that lead to potential cross-resistance between organophosphate and pyrethroids are discussed. PMID- 19556576 TI - Analysis of childhood morbidity with geoadditive probit and latent variable model: a case study for Egypt. AB - This work applies geoadditive latent variable models to analyze the impact of risk factors and the spatial effects on the latent, unobservable variable "health status" or "frailty" of a child less than 5 years of age using the 2003 Demographic and Health survey (DHS) data from Egypt. Childhood diseases are a major cause of death of children in the developing world. In developing countries a quarter of infant and childhood mortality is related to childhood disease, particularly to diarrhea. Our case study is based on the 2003 Demographic and Health Survey for Egypt (EDHS). It provided data on the prevalence and treatment of common childhood disease such as diarrhea, cough, and fever, which are seen as symptoms or indicators of children's health status, causing increased morbidity and mortality. These causes are often associated with a number of risk factors, including inadequate antenatal care, lack of or inadequate vaccination, and environmental factors that affected the health of the child in early years, various bio-demographic and socioeconomic variables. In this work, we investigate the impact of such factors on childhood disease with flexible geoadditive models. These models allow us to analyze usual linear effects of covariates, nonlinear effects of continuous covariates, and small-area regional effects within a unified, semi-parametric Bayesian framework for modeling and inference. As a first step, we use separate geoadditive probit models the binary target variables for diarrhea, cough, and fever using covariate information from the EDHS. Based on these results, we then apply recently developed geoadditive latent variable models where the three observable disease variables are taken as indicators for the latent individual variable "health status" or "frailty" of a child. This modeling approach allows us to study the common influence of risk factors on individual frailties of children, thereby automatically accounting for association between diseases as indicators for health status. PMID- 19556577 TI - Antibiotic use in pediatric patients admitted to a referral hospital in Botswana. AB - Inappropriate antimicrobial drug use is well described for hospitalized patients in the United States. Antibiotic use in hospitals in developing countries is less well documented. We evaluated the antibiotics prescribed to 91 pediatric inpatients in Botswana. The results showed that the duration of prescribed therapy can be excessive. Recommendations for potential interventions to reduce antibiotic overuse in this setting are necessary. PMID- 19556578 TI - Efficacy of protocols for cleaning and disinfecting infant feeding bottles in less developed communities. AB - Although breastfeeding is the best choice for most infants, infant formula is used widely, commonly introduced during the neonatal period, and usually given to infants in bottles that can be difficult to clean. We artificially contaminated infant feeding bottles with low and high inocula of bacterial enteric pathogens and evaluated the efficacy of several cleaning and chlorine disinfection protocols. Rinsing with soapy water followed by tap water was the most effective cleaning method and reduced pathogen load by 3.7 and 3.1 log(10)s at the low and high inoculum levels, respectively. Submersion in 50 ppm hypochlorite solution for 30 minutes produced a 3.7-log(10) reduction in pathogens, resulting in no identifiable pathogens among bottles. This result was comparable to boiling. When combined with handwashing, use of safe water, and appropriate storage of prepared infant formula, these simple, inexpensive practices could improve the microbiological safety of infant formula feeding in less developed settings. PMID- 19556579 TI - Difficulties in maintaining improved handwashing behavior, Karachi, Pakistan. AB - In an earlier study in Karachi, Pakistan, households that received free soap and handwashing promotion for 9 months reported 53% less diarrhea than controls. Eighteen months after the intervention ended, these households were enrolled in a follow-up study to assess sustainability of handwashing behavior. Upon re enrollment, mothers in households originally assigned to the intervention were 1.5 times more likely to have a place with soap and water to wash hands (79% versus 53%, P = 0.001) and when asked to wash hands were 2.2 times more likely to rub their hands together at least three times (50% versus 23%, P = 0.002) compared with controls. In the ensuing 14 months, former intervention households reported a similar proportion of person-days with diarrhea (1.59% versus 1.88%, P = 0.66) as controls. Although intervention households showed better handwashing technique after 2 years without intervention, their soap purchases and diarrhea experience was not significantly different from controls. PMID- 19556580 TI - Etiology of acute undifferentiated febrile illness in the Amazon basin of Ecuador. AB - We conducted a longitudinal observational study of 533 patients presenting to two hospitals in the Ecuadorean Amazon basin with acute undifferentiated febrile illness (AUFI) from 2001 through 2004. Viral isolation, reverse transcription polymerase chain reaction (RT-PCR), IgM seroconversion, and malaria smears identified pathogens responsible for fever in 122 (40.1%) of 304 patients who provided both acute and convalescent blood samples. Leptospirosis was found in 40 (13.2%), malaria in 38 (12.5%), rickettsioses in 18 (5.9%), dengue fever in 16 (5.3%), Q fever in 15 (4.9%), brucellosis in 4 (1.3%), Ilheus infection in 3 (1.0%), and Venezuelan equine encephalitis (VEE), Oropouche, and St. Louis encephalitis virus infections in less than 1% of these patients. Viral isolation and RT-PCR on another 229 participants who provided only acute samples identified 3 cases of dengue fever, 2 of VEE, and 1 of Ilheus. None of these pathogens, except for malaria, had previously been detected in the study area. PMID- 19556581 TI - Incidence and risk factors of Opisthorchis viverrini infections in a rural community in Thailand. AB - Opisthorchis viverrini infection is one of the major public health problems in Thailand. Although information about transmission of this parasite to humans is well established, infections are still prevalent in the northern and northeastern Thailand. Thus, prospective epidemiologic information is needed for developing more effective public health interventions. A cohort study to identify incidence and risk factors of O. viverrini infection was conducted in a rural community in Thailand during December 2002-February 2004. The incidence rate of opisthorchiasis was 21.6/100 person-years. The independent factors associated with opisthorchiasis were an age > 60 years and consuming chopped raw fish salad (Koi pla). Thus, avoiding this raw fish salad should be emphasized in the national control program. PMID- 19556582 TI - MM3-ELISA detection of Fasciola hepatica coproantigens in preserved human stool samples. AB - In this study, we evaluate the MM3-COPRO method for detection of Fasciola coproantigens in human fecal samples, and the usefulness of a new preservative/diluent, CoproGuard, developed for preservation of Fasciola coproantigens. The MM3-COPRO assay was evaluated with 213 samples from healthy patients, 30 Fasciola positive fecal samples (according to the Kato-Katz method), and 83 samples from patients with other parasitic infections. All Fasciola positive specimens were detected with the MM3-COPRO assay (100% sensitivity) and there was no cross-reactivity with other common parasites present in the clinical specimens analyzed (100% specificity). The use of CoproGuard enhanced coproantigen extraction without affecting the detection limit of the assay, and the antigenicity of Fasciola coproantigens in fecal samples stored at 37 degrees C was retained throughout the entire observation period (120 days). We concluded that the MM3-COPRO ELISA combined with the use of CoproGuard may be a very useful tool for the diagnosis of human fascioliasis. PMID- 19556583 TI - Dose-response assay templates for in vitro assessment of resistance to benzimidazole and nicotinic acetylcholine receptor agonist drugs in human hookworms. AB - With the implementation of mass drug administration programs for the control of human hookworms, there is a need to monitor for the emergence of drug resistance. We have therefore examined in vitro assays for monitoring sensitivity to benzimidazoles (egg hatch assay) and nicotinic acetylcholine receptor agonist drugs (motility and morphology assays), with a view to developing tools for monitoring drug sensitivity in the field. We have performed assays with Necator americanus and Ancylostoma ceylanicum, and combined this with published data on N. americanus and Ancylostoma caninum, to indicate the breadth of the responses of various hookworm species and isolates in these in vitro assays. This has allowed us to generate assay templates covering the known range of responses, with scope to cover any shift in response that may be indicative of resistance. These assays will have immediate applicability in monitoring for the emergence of drug resistance in human hookworm populations. PMID- 19556584 TI - A community-based survey of human toxoplasmosis in rural Amazonia: seroprevalence, seroconversion rate, and associated risk factors. AB - IgG antibodies to Toxoplasma gondii were detected in, March-April 2004, in 65.8% (95% confidence interval, 60.8-70.8%) of 342 systematically sampled subjects 5-90 years of age (87.5% of the eligible) living in a rural settlement in Amazonia, with a seroconversion rate of 9% over 1 year of follow-up of 99 seronegative subjects. Multiple logistic regression analysis identified age as the only significant independent predictor of seropositivity at the baseline. Each additional year of age increases the odds of being seropositive by 6%, and 76.8% of the subjects are expected to be seropositive at 30 years of age. A single high prevalence spatial cluster, comprising 11.9% of the seropositive subjects, was detected in the area; households in the cluster were less likely to have dogs as pets and their heads had a lower education level, when compared with households located outside the cluster. The challenges for preventing human toxoplasmosis in tropical rural settings are discussed. PMID- 19556585 TI - Changes in West Nile virus seroprevalence and antibody titers among Wisconsin mesopredators 2003-2006. AB - After the 2001 occurrence of West Nile virus (WNV) in Wisconsin (WI), we collected sera, during 2003-2006, from south-central WI mesopredators. We tested these sera to determine WNV antibody prevalence and geometric mean antibody titer (GMAT). Four-fold higher antibody prevalence and 2-fold higher GMAT in 2003-2004 indicated greater exposure of mesopredators to WNV during the apparent epizootic phase. The period 2005-2006 was likely the enzootic phase because WNV antibody prevalence fell to a level similar to other flaviviruses. Our results suggest that, in mesopredators, vector-borne transmission is the primary route of infection and WNV antibodies persist for < 1 year. Mesopredators may be sensitive indicators of West Nile virus spill-over into humans and horses. Mesopredator sero-surveys may complement dead crow surveillance by providing additional data for the timing of public health interventions. Research is needed to clarify the dynamics of WNV infection in these mammals and their role as potential WNV amplifiers. PMID- 19556586 TI - Seasonality of influenza in the tropics: a distinct pattern in northeastern Brazil. AB - Influenza epidemics occur worldwide annually. The incidence of influenza shows a seasonal pattern in temperate areas, but little is known about influenza seasonality in tropical regions. The objective of this study was to determine the prevalence and the seasonal pattern of influenza infections in children living in the city of Fortaleza in northeastern Brazil. An indirect immunofluorescence assay was performed on nasopharyngeal aspirates collected from children attending in ambulatories, emergency rooms, and wards of the Hospital Infantil Albert Sabin with suspicion of acute respiratory infection during 7 consecutive years (2001 2007). Influenza viruses were detected in 6.3% (234/3,708) of specimens. Laboratory-based surveillance data showed a clear annual epidemic cycle of influenza, with a peak usually occurring in the rainy periods. In Fortaleza, flu infections occurred at a low level throughout the year but exhibit a marked seasonal increase during the rainy season. PMID- 19556588 TI - Biological bulletin virtual symposium: biology of marine invertebrate larvae. PMID- 19556587 TI - Lack of evidence of hepatitis C and HIV co-infection among men who have sex with men in Peru. AB - Hepatitis C virus (HCV) infection occurs among human immunodeficiency virus (HIV) infected men who have sex with men (MSM) because of shared routes of transmission. To assess the association between HCV and HIV infection among MSM in Peru, we conducted a matched case-control study (162 HIV-positive cases and 324 HIV-negative controls) among participants of an HIV sentinel surveillance survey in six urban cities. The HCV infection was initially screened using anti HCV ELISA and immunoblot assay, and thereafter confirmed by the HCV RNA qualitative assay. Among cases, no confirmed HCV infection was found while among controls, only two confirmed HCV infections were reported (0.62%). This matched case-control reports a very low probability of association between HCV and HIV co infection and suggests a very low prevalence of HCV infection among MSM in Peru. PMID- 19556589 TI - How did indirect development with planktotrophic larvae evolve? AB - The two main types of theories for the evolution of the biphasic life cycles in marine invertebrates are discussed. The "intercalation" theories propose that the larval stages (planktotrophic or lecithotrophic) have evolved as specializations from the ancestral, direct life cycle. The opposing "terminal addition" theories propose that the ancestor was holopelagic and that the adult stage was added to the life cycle with the pelagic stage retained as a planktotrophic larva. It is emphasized that theories based on hypothetical ancestors that were unable to feed must be rejected. This applies to planula theories based on a compact planula. Various arguments against the theories that consider the feeding larvae as ancestral in the major eumetazoan lineages and in particular against the trochaea theory are discussed and found untenable. It is suggested that the "Cambrian explosion" was actually a rapid Ediacaran radiation of the eubilaterians that was made possible by the evolution of a tubular gut with all the resulting possibilities for new body plans. PMID- 19556590 TI - Molluscan larvae: Pelagic juveniles or slowly metamorphosing larvae? AB - Asking the right questions about evolution of development, larval morphology, and life history requires knowledge of ancestral state. Two hypotheses dominate current opinion about the ancestral life cycle of bilaterians: the "larva-first" and the "intercalation" hypotheses. Until recently, the larva-first hypothesis was preeminent. This proposes that the original indirect life cycle of bilaterians included a planktotrophic larva followed by a benthic adult. Phylogenetic evidence suggests that a planktotrophic larva is plesiomorphic for echinoderms. A preponderance of developmental studies on echinoderms may have fostered a tendency to extrapolate conclusions about echinoderm development to other clades, particularly the concept that larval and juvenile/adult bodies are mostly separate entities. However, some of the recent reconstructions of bilaterian phylogeny suggest that nonfeeding larvae may have been ancestral for bilaterians, and these may have been intercalated into a life cycle that was originally direct. I review comparative data on molluscan development that suggests the trochophore-like stage is little more than a gastrula with transient structures (prototroch and apical sensory organ) to allow a temporary planktonic phase during development. Most lineage founder cells of molluscan embryos generate progeny that develop through the veliger stage into structures of the juvenile, which becomes benthic when the prototroch and apical sensory organ are lost. In light of this, the model of separate larval and juvenile bodies with the latter developing from nests of multipotent cells within the larva is inappropriate for molluscs. The intercalation hypothesis may be a better model for interpreting development of molluscs and other lophotrochozoans. PMID- 19556591 TI - Egg size as a life history character of marine invertebrates: Is it all it's cracked up to be? AB - Egg size is one of the most important aspects of the life history of free spawning marine organisms, and it is correlated with larval developmental mode and many other life-history characters. Egg size is simple to measure and data are available for a wide range of taxa, but we have a limited understanding of how large and small eggs differ in composition; size is not always the best measure of the characters under selection. Large eggs are generally considered to reflect increased maternal investment, but egg size alone can be a poor predictor of energetic content within and among taxa. We review techniques that have been used to measure the energetic content and biochemical makeup of invertebrate eggs and point out the strengths and difficulties associated with each. We also suggest a number of comparative and descriptive approaches to biochemical constituent analysis that would strengthen our understanding of how natural selection shapes oogenic strategies. Finally, we highlight recent empirical research on the intrinsic factors that drive intraspecific variation in egg size. We also highlight the relative paucity of these data in the literature and provide some suggestions for future research directions. PMID- 19556592 TI - Larval biology of the crab Rhithropanopeus harrisii (Gould): a synthesis. AB - This synthesis reviews the physiological ecology and behavior of larvae of the benthic crab Rhithropanopeus harrisii, which occurs in low-salinity areas of estuaries. Larvae are released rhythmically around the time of high tide in tidal estuaries and in the 2-h interval after sunset in nontidal estuaries. As in most subtidal crustaceans, the timing of larval release is controlled by the developing embryos, which release peptide pheromones that stimulate larval release behavior by the female to synchronize the time of egg hatching. Larvae pass through four zoeal stages and a postlarval or megalopal stage that are planktonic before metamorphosis. They are retained near the adult population by means of an endogenous tidal rhythm in vertical migration. Larvae have several safeguards against predation: they undergo nocturnal diel vertical migration (DVM) and have a shadow response to avoid encountering predators, and they bear long spines as a deterrent. Photoresponses during DVM and the shadow response are enhanced by exposure to chemical cues from the mucus of predator fishes and ctenophores. The primary visual pigment has a spectral sensitivity maximum at about 500 nm, which is typical for zooplankton and matches the ambient spectrum at twilight. Larvae can detect vertical gradients in temperature, salinity, and hydrostatic pressure, which are used for depth regulation and avoidance of adverse environmental conditions. Characteristics that are related to the larval habitat and are common to other crab larval species are considered. PMID- 19556593 TI - Quantifying the "bio-" components in biophysical models of larval transport in marine benthic invertebrates: advances and pitfalls. AB - Biophysical models are being used increasingly, both as predictive tools of larval dispersal for a particular system and for general evaluation of the role of different factors in larval transport. In the results of such models, larval duration, mortality, and behavior in the water column have exhibited pronounced effects on larval dispersal of marine benthic invertebrates. The parameterization of these processes has broadly reflected values from laboratory experiments, but the accuracy of these values is unknown. The pelagic larval duration used in models should be determined by laboratory, or preferably field, studies and should incorporate environmentally dependent variability. For mortality, in situ estimates are now feasible and, likely, more accurate than the currently used values. Larval behavior can be measured in the field, by high-frequency sampling of distributional changes relative to features in the water column or by controlled larval releases in tractable systems. To successfully validate the outcomes of these models, we must either improve our techniques for measuring larval abundance at the end of larval transport immediately before settlement, or incorporate components for settlement into the models. We must also address the mismatch in sampling resolution between biological and physical processes. If used with caution, this powerful approach can significantly advance our understanding of larval transport. PMID- 19556594 TI - Larval development with transitory epidermis in Paranemertes peregrina and other hoplonemerteans. AB - We describe development of the hoplonemertean Paranemertes peregrina from fertilization to juvenile, using light, confocal, and electron microscopy. We discovered that the uniformly ciliated lecithotrophic larva of this species has a transitory epidermis, which is gradually replaced by the definitive epidermis during the course of planktonic development. The approximately 90 large multiciliated cleavage-arrested cells of the transitory larval epidermis become separated from each other by intercalating cells of the definitive epidermis, then gradually diminish in size and disappear more or less simultaneously. Rudiments of all major adult structures-the gut, proboscis, cerebral ganglia, lateral nerve cords, and cerebral organs-are already present in 4-day-old larvae. Replacement of the epidermis is the only overt metamorphic transformation of larval tissue; larval structures otherwise prefigure the juvenile body, which is complete in about 10 days at 7-10 degrees C. Our findings on development of digestive system, nervous system, and proboscis differ in several ways from previous descriptions of hoplonemertean development. We report development with transitory epidermis in two other species, review evidence from the literature, and suggest that this developmental type is the rule for hoplonemerteans. The hoplonemertean planuliform larva is fundamentally different both from the pilidium larva of the sister group to the Hoplonemertea, the Pilidiophora, and from the hidden trochophore of palaeonemerteans. We discuss the possible function and homology of the larval epidermis in development of other nemerteans and spiralians in general. PMID- 19556595 TI - Shaping the things to come: ontogeny of lophotrochozoan neuromuscular systems and the tetraneuralia concept. AB - Despite the large variation in adult bodyplan phenotypes, a worm-shaped morphology is considered plesiomorphic for both Lophotrochozoa and Bilateria. Although almost all larval and adult lophotrochozoan worms have serially arranged ring muscles in their body wall, a comparison of their ontogeny reveals no less than six different developmental pathways that lead to this homogenous arrangement of ring muscles. However, in all taxa, with the exception of chaetodermomorph molluscs and the segmented annelids, ring muscle development starts with synchronous formation of certain pioneer myocytes, which is thus considered basal for Lophotrochozoa. Recent studies on spiralian neurogenesis revealed remnants of ancestral segmentation in echiurans and sipunculans, thus confirming molecular phylogenetic studies that propose a close relationship of these three taxa. Larval entoprocts exhibit a mosaic of larval and adult molluscan characters and, among other apomorphies, share with polyplacophoran Mollusca a complex larval apical organ and a tetraneurous nervous system, strongly suggesting a monophyletic assemblage of Entoprocta and Mollusca. The term Tetraneuralia is proposed herein for this lophotrochozoan clade. Overall, formation of the lophotrochozoan neuromuscular bodyplan appears as a highly dynamic process on both the ontogenetic and the evolutionary timescales, highlighting the importance of insights into these processes for reconstructing ancestral bodyplan features and phylogenetic relationships. PMID- 19556596 TI - Sniffing out new data and hypotheses on the form, function, and evolution of the echinopluteus post-oral vibratile lobe. AB - The performance requirements of ciliary band feeding explain the convoluted forms of many marine invertebrate larvae. Convolutions increase surface area and therefore feeding rates per unit body volume. We review recent advances in morphology, neural development, and behavior at settlement of the echinoid Lytechinus pictus and provide new ultrastructural and expression data on larvae of its congener, L. variegatus. Larvae of the echinometrid Colobocentrotus atratus contain neurons identified by their expression of nitric oxide synthase (NOS), indicating that this character is not unique to Lytechinus. We hypothesize that in some echinoids the convoluted shape of the post-oral vibratile lobe (POVL) covaries with the distribution of identified sensory neurons to enable olfaction during settlement. An analysis of variation in structural elaboration of the post-oral transverse ciliary band (PTB) within Echinoida and in feeding larvae of other echinoderm classes indicates that only echinoids, but not all echinoids, possess this novel character; larvae that do are distributed heterogeneously within the class. In recognition of this specialized function for the POVL and surrounding ectoderm, and because it is lobate and grows toward the mouth, we propose naming this structure the adoral lobe. PMID- 19556597 TI - Nervous system development in feeding and nonfeeding asteroid larvae and the early juvenile. AB - Larval and juvenile nervous systems (NS) of three asterinid sea stars with contrasting feeding and nonfeeding modes of development were characterized using the echinoderm-specific synaptotagmin antibody. In the feeding bipinnaria and brachiolaria larvae of Patiriella regularis, the species with ancestral-type development, an extensive NS was associated with the ciliary bands (CBs) and attachment complex. Lecithotrophic planktonic (Meridastra calcar) and benthic (Parvulastra exigua) brachiolariae lacked CBs and the associated NS, but had an extensive NS in the attachment complex. The similarity in the distribution and morphology of synaptotagmin immunoreactive neurons and the anatomy of the NS in the attachment complex of these closely related sea stars suggests conservation of neurogenesis in settlement-stage larvae regardless of larval feeding mode. Nerve cells were prominent on the brachia of all three species. In advanced brachiolariae the larval nervous system was localized to the adhesive disc as the larval body resorbed during metamorphosis. The structures and tissues that contained larval neurons degenerated during metamorphosis. There was no evidence that the larval NS persists through metamorphosis. In juvenile development, synaptotagmin IR was first evident in the NS of the tube feet. As the central nervous system developed, synaptotagmin IR reflected the histological organization of the adult NS. The juvenile NS formed de novo with a temporal lapse between histogenesis and synaptotagmin IR. We evaluated the ontogeny of NS organization in the change in body plan from the bilateral larva to the radial juvenile. PMID- 19556598 TI - Feeding ability of early zoeal stages of the Norway lobster Nephrops norvegicus (L.). AB - The wide geographical distribution of the Norway lobster, Nephrops norvegicus, results in a delay, with latitudinal decrease, in the larval season from spring to winter. Newly hatched larvae of the species may therefore be exposed to suboptimal levels or types of prey and face intermittent periods of starvation at low latitudes. This work investigated the feeding response of the first two zoeal stages of N. norvegicus under variable prey densities, prey types, feeding histories, and photoperiods. Both zoeae (Z) I and II increased the number of consumed prey with increasing food levels. ZI preferred Artemia sp. nauplii over larger metanauplii, while in ZII, higher ingestion was observed only for metanauplii at higher food concentrations. The number of prey ingested by larvae previously starved or under low food conditions was always higher than that of larvae exposed to high food levels. These findings seem to indicate that larvae may maximize prey ingestion in the presence of plankton patches with higher food abundance and minimize the deleterious effects induced by previous periods of intermittent starvation or unsuitable prey densities or types. Extreme photoperiods (24 and 0 h of light) did not improve larval feeding ability and are not a suitable option for larviculture. PMID- 19556599 TI - Pre-settlement behavior in larval bryozoans: the roles of larval age and size. AB - Larval behaviors prior to settlement are important for both dispersal and the likelihood that larvae will encounter settlement habitat. The role of endogenous factors such as larval age and size are likely to be important in determining pre settlement behavior but are less well understood than exogenous factors. In a simple experiment we explored the role of larval age and size on pre-settlement behavior in two species of bryozoan. We then used the results of this experiment to develop a theoretical model, which explored potential fitness benefits associated with phenotype-dependent changes in larval behavior (i.e., behaviors that changed depending on larval age or larval size) in a heterogeneous environment. In the experiment we delayed the metamorphosis of larvae of Bugula neritina andWatersipora arcuata and assessed the changes in the behavior of individual larvae (exploring the substratum vs. swimming away from it) as a function of larval age and size. In B. neritina, larval size had no effect on larval swimming behavior, but the youngest and oldest larvae spent more time exploring the substrate than did larvae of intermediate age. In W. arcuata, larval size and age had interactive effects on larval behavior. Our theoretical model predicted that phenotype-dependent behaviors carried a fitness benefit relative to phenotype-independent behaviors, but this depended strongly on the availability and quality of habitat elsewhere. We suggest that, taken together, larval age and size are important endogenous factors that act to affect pre settlement larval behavior and that changes in behavior may act to increase fitness. PMID- 19556600 TI - Not my "type": larval dispersal dimorphisms and bet-hedging in opisthobranch life histories. AB - When conditions fluctuate unpredictably, selection may favor bet-hedging strategies that vary offspring characteristics to avoid reproductive wipe-outs in bad seasons. For many marine gastropods, the dispersal potential of offspring reflects both maternal effects (egg size, egg mass properties) and larval traits (development rate, habitat choice). I present data for eight sea slugs in the genus Elysia (Opisthobranchia: Sacoglossa), highlighting potentially adaptive variation in traits like offspring size, timing of metamorphosis, hatching behavior, and settlement response. Elysia zuleicae produced both planktotrophic and lecithotrophic larvae, a true case of poecilogony. Both intracapsular and post-hatching metamorphosis occurred among clutches of "Boselia" marcusi, E. cornigera, and E. crispata, a dispersal dimorphism often misinterpreted as poecilogony. Egg masses of E. tuca hatched for up to 16 days but larvae settled only on the adult host alga Halimeda, whereas most larvae of E. papillosa spontaneously metamorphosed 5-7 days after hatching. Investment in extra-capsular yolk may allow mothers to increase larval size relative to egg size and vary offspring size within and among clutches. Flexible strategies of larval dispersal and offspring provisioning in Elysia spp. may represent adaptations to the patchy habitat of these specialized herbivores, highlighting the evolutionary importance of variation in a range of life-history traits. PMID- 19556601 TI - Pelagic larval duration and dispersal distance revisited. AB - I present dispersal distances for 44 species with data on propagule duration (PD) for 40 of these. Data were combined with those in Shanks et al. (2003; Ecol. Appl. 13: S159-S169), providing information on 67 species. PD and dispersal distance are correlated, but with many exceptions. The distribution of dispersal distances was bimodal. Many species with PDs longer than 1 day dispersed less than 1 km, while others dispersed tens to hundreds of kilometers. Organisms with short dispersal distances were pelagic briefly or remained close to the bottom while pelagic. Null models of passively dispersing propagules adequately predict dispersal distance for organisms with short PDs (<1 day), but overestimate dispersal distances for those with longer PDs. These models predict that propagules are transported tens of kilometers offshore; however, many types remain within the coastal boundary layer where currents are slower and more variable, leading to lower than predicted dispersal. At short PDs, dispersal distances estimated from genetic data are similar to observed. At long PDs, genetic data generally overestimate dispersal distance. This discrepancy is probably due to the effect of rare individuals that disperse long distances, thus smoothing genetic differences between populations. Larval behavior and species' life-history traits can play a critical role in determining dispersal distance. PMID- 19556602 TI - Systemic administration of sphingosine-1-phosphate increases bronchial hyperresponsiveness in the mouse. AB - Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid that plays important roles in allergic responses, including asthma. S1P acts on many cell types, such as mast cells, the airway epithelium, airway smooth muscle, and many immune cells. In this study we have evaluated whether a systemic administration of S1P to Balb/c mice modifies airway reactivity. Our data show that S1P (0.1-10 ng) given subcutaneously to Balb/c mice causes a specific and dose-dependent increase in cholinergic reactivity of bronchial tissues in vitro. This effect is (1) dose dependent, with a maximal effect of the dose of 10 ng of S1P; and (2) time dependent, reaching a maximal effect 21 days after S1P administration. Similarly, in the whole lung assay there is a dose- and time-dependent increase in lung resistance. Lungs isolated from S1P-treated mice displayed an increase in mast cell number. Furthermore, there is an increase of IL-4, IL-13, and IL-17 production. In conclusion, our data demonstrate that S1P signaling is involved in the complex pathway underlying airway hyperresponsiveness. PMID- 19556603 TI - Nitric oxide-mediated bcl-2 stabilization potentiates malignant transformation of human lung epithelial cells. AB - Hexavalent chromium (Cr(VI)) compounds are known human carcinogens associated with the incidence of lung cancer. Although a direct correlation between Cr(VI) exposure and lung cancer has been established, several studies aimed at generating animal models for Cr(VI) have yielded inconsistent data that do not affirmatively support findings from epidemiologic studies. Because the lack of a good animal model has hindered the identification of molecular mechanisms involved in Cr(VI) exposure, we developed an in vitro model that facilitates mechanistic studies of Cr(VI)-induced carcinogenesis. We report here that long term exposure to Cr(VI) leads to the malignant transformation of nontumorigenic human lung epithelial cells. Cr(VI)-transformed cells exhibited loss of contact inhibition, colony formation, and increased rates of cell invasion, migration, and proliferation, as compared with passage-matched control cells. Cr(VI) transformed cells evaded apoptosis by a mechanism involving S-nitrosylation and stabilization of Bcl-2 protein in a nitric oxide-dependent manner. This study establishes an important in vitro model that facilitates mechanistic studies of Cr(VI)-induced carcinogenesis, and elucidates a novel mechanism that causes apoptosis-resistant malignant transformation of nontumorigenic lung cells in response to a human carcinogen. PMID- 19556605 TI - Regulation of neuregulin 1beta1-induced MUC5AC and MUC5B expression in human airway epithelium. AB - Excessive mucus production has been linked to many of the pathologic features of respiratory diseases, including obstruction of the airways, decline in lung function, increased rates of mortality, and increased infections. The mucins, MUC5AC and MUC5B, contribute to the viscoelastic properties of mucus, and are found at elevated levels in the airways of individuals with chronic respiratory diseases. The T helper type 2 cell cytokine, IL-13, is known to regulate MUC5AC expression in goblet cells of the airways, although much less is known about the regulation of MUC5B expression. In a study to further understand the mediators of MUC5AC and MUC5B expression, neuregulin (NRG) 1beta1 was identified as novel regulator of goblet cell formation in primary cultures of human bronchial epithelial cells (HBECs). NRG1beta1 increased expression of MUCAC and MUC5B proteins in a time- and dose-dependent fashion in HBEC cultures. NRG1beta1 induced expression of MU5AC and MUC5B was shown to involve v-erb-b2 erythroblastic leukemia viral oncogene homolog (ErbB) and ErbB3 receptors, but not ErbB4 receptors. Treatment of HBECs with inhibitors of p38 mitogen-activated protein kinase, extracellular signal-regulated kinase1/2, and phosphatidylinositol 3-kinase indicated that these kinases were involved in NRG1beta1-induced MUC5AC and MUC5B expression. Additionally, NRG1beta1 was shown to induce the phosphorylation of the ErbB2 receptor, AKT, and extracellular signal-regulated kinase 1/2. NRG1beta1 protein was found increased in the airways of antigen-challenged mice, together with increases in MUC5AC and MUC5B message. Together, these data indicate that NRG1beta1 is a novel mediator of MUC5AC and MUC5B expression in HBECs, and may represent a novel therapeutic target for mucus hypersecretion in respiratory diseases. PMID- 19556604 TI - Sex differences in estrogen receptor subcellular location and activity in lung adenocarcinoma cells. AB - The role of estrogens in the increased risk of lung adenocarcinoma in women remains uncertain. We reported that lung adenocarcinoma cell lines from female, but not male, patients with non-small cell lung cancer respond proliferatively and transcriptionally to estradiol (E(2)), despite equal protein expression of estrogen receptors (ER) alpha and beta. To test the hypothesis that nuclear localization of ER alpha corresponds to genomic E(2) activity in lung adenocarcinoma cells from females, cell fractionation, immunoblot, and confocal immunohistochemical microscopy were performed. We report for the first time that E(2) increases phospho-serine-118-ER alpha (P-ser118-ER alpha) and cyclin D1 (CCND1) nuclear colocalization in H1793, but not A549 lung adenocarcinoma cells, derived from a female and male patient, respectively. ER beta was primarily in the cytoplasm and mitochondria, independent of E(2) treatment, and showed no difference between H1793 and A549 cells. E(2) induced higher transcription of endogenous ER alpha-regulated CCND1 in H1793 than in A549 cells. Likewise, higher rapid, non-genomic E(2)-induced extracellular signal-regulated kinase 1/2 activation was detected in H1793 compared with A549 cells, linking extracellular signal-regulated kinase activation to increased P-ser118-ER alpha. Furthermore, E(2) increased cyclin D1 and P-ser118-ER alpha nuclear localization in H1793, but not A549 cells. Together, our results indicate that nuclear localization of P ser118-ER alpha provides one explanation for sex-dependent differences in E(2) genomic responses in lung adenocarcinoma cell lines. PMID- 19556607 TI - ABCD2 is abundant in adipose tissue and opposes the accumulation of dietary erucic acid (C22:1) in fat. AB - The ATP binding cassette transporter, ABCD2 (D2), is a peroxisomal protein whose mRNA has been detected in the adrenal, brain, liver, and fat. Although the role of this transporter in neural tissues has been studied, its function in adipose tissue remains unexplored. The level of immunoreactive D2 in epididymal fat is >50-fold of that found in brain or adrenal. D2 is highly enriched in adipocytes and is upregulated during adipogenesis but is not essential for adipocyte differentiation or lipid accumulation in day 13.5 mouse embryonic fibroblasts isolated from D2-deficient (D2(-/-)) mice. Although no differences were appreciated in differentiation percentage, total lipid accumulation was greater in D2(-/-) adipocytes compared with the wild type. These results were consistent with in vivo observations in which no significant differences in adiposity or adipocyte diameter between wild-type and D2(-/-) mice were observed. D2(-/-) adipose tissue showed an increase in the abundance of 20:1 and 22:1 fatty acids. When mice were challenged with a diet enriched in erucic acid (22:1), this lipid accumulated in the adipose tissue in a gene-dosage-dependent manner. In conclusion, D2 is a sterol regulatory element binding protein target gene that is highly abundant in fat and opposes the accumulation of dietary lipids generally absent from the triglyceride storage pool within adipose tissue. PMID- 19556606 TI - Attenuation of chronic pulmonary inflammation in A2B adenosine receptor knockout mice. AB - Pharmacologic evidence suggests that activation of A(2B) adenosine receptors results in proinflammatory effects relevant to the progression of asthma, a chronic lung disease associated with elevated interstitial adenosine concentrations in the lung. This concept has been challenged by the finding that genetic removal of A(2B) receptors leads to exaggerated responses in models of acute inflammation. Therefore, the goal of our study was to determine the effects of A(2B) receptor gene ablation in the context of ovalbumin-induced chronic pulmonary inflammation. We found that repetitive airway allergen challenge induced a significant increase in adenosine levels in fluid recovered by bronchoalveolar lavage. Genetic ablation of A(2B) receptors significantly attenuated allergen-induced chronic pulmonary inflammation, as evidenced by a reduction in the number of bronchoalveolar lavage eosinophils and in peribronchial eosinophilic infiltration. The most striking difference in the pulmonary inflammation induced in A(2B) receptor knockout (A(2B)KO) and wild-type mice was the lack of allergen-induced IL-4 release in the airways of A(2B)KO animals, in line with a significant reduction in IL-4 protein and mRNA levels in lung tissue. In addition, attenuation of allergen-induced transforming growth factor-beta release in airways of A(2B)KO mice correlated with reduced airway smooth muscle and goblet cell hyperplasia/hypertrophy. In conclusion, genetic removal of A(2B) adenosine receptors in mice leads to inhibition of allergen induced chronic pulmonary inflammation and airway remodeling. These findings are in agreement with previous pharmacologic studies suggesting a deleterious role for A(2B) receptor signaling in chronic lung inflammation. PMID- 19556608 TI - Feeding jejunostomy for the treatment of severe hyperemesis gravidarum: a case series. AB - BACKGROUND: Hyperemesis gravidarum is severe nausea and vomiting during pregnancy leading to dehydration, nutrition deficiency, and fetal morbidity and mortality. Treatment must maintain fluid and electrolyte balance and caloric intake. Parenteral nutrition is often attempted; however, complication rates are high. Nutrition via nasoenteric and percutaneous endoscopic gastrostomy tubes is limited by poor patient tolerance, tube dislodgement, and altered anatomy in pregnancy. METHODS: Women with hyperemesis gravidarum who failed standard therapy were offered jejunostomy. All patients underwent surgical jejunostomy in the second trimester. Isotonic tube feeds were administered to a goal caloric factor calculated by the Harris-Benedict equation with a correction added for pregnancy. Patients were monitored until delivery. RESULTS: Five women underwent jejunostomy placement at our institution between 1998 and 2005. One patient underwent jejunostomy placement twice for consecutive pregnancies. The mean body weight loss from prepregnancy was 7.9% (range, 4.0%-15.9%). Patients underwent jejunostomy placement between 12 and 26 weeks of gestation (median 14 weeks). Twelve to 16 Fr catheters were placed in the proximal jejunum. Maternal weight gain occured in 5 of 6 pregnancies. The mean duration of tube placement was 19 weeks (range, 8-28 weeks). All pregnancies ended with term deliveries (range, 36 40 weeks of gestation). The mean infant birth weight was 2885 g (range, 2270-4000 g). Tube-related complications were limited to dislodgement in 2 patients in the third trimester. No cases of infection, bleeding, or preterm labor occured. CONCLUSIONS: Feeding via jejunostomy is a potentially safe, effective, and well tolerated mode of nutrition support therapy in hyperemesis gravidarum. PMID- 19556610 TI - Use of probiotics for treatment of chemotherapy-induced diarrhea: is it a myth? PMID- 19556609 TI - Parenteral feeding depletes pulmonary lymphocyte populations. AB - BACKGROUND: The effect of parenteral nutrition (PN) on lymphocyte mass in the lung is unknown, but reduced mucosal lymphocytes are hypothesized to play a role in the reduced immunoglobulin A-mediated immunity in both gut and lung. The ability to transfer and track cells between mice may allow study of diet-induced mucosal immune function. The objectives of this study are to characterize lung T cell populations following parenteral feeding and to study distribution patterns of transferred donor lung T cells in recipient mice. METHODS: In experiment 1, cannulated male Balb/c mice are randomized to receive chow or PN for 5 days. Lung lymphocytes are obtained via collagenase digestion, and flow cytometric analysis is used to identify total T (CD3+) and B (CD45/B220+) cells. In experiment 2, isolated lung T cells from chow-fed male Balb/c mice are pooled and labeled in vitro with a fluorescent dye (carboxyfluorescein diacetate succinimidyl ester [CFSE]), and 1.1 x 10(8) CFSE+ cells (3.1 x 10(6) T cells) are transferred to chow-fed Balb/c recipients. Cells recovered from recipient lungs and intestinal lamina propria (LP) are analyzed by flow cytometry to determine CFSE/CD3+ T cells at 1, 2, and 7 days. In experiment 3, cells are transferred to PN-fed recipients. RESULTS: In experiment 1, PN significantly decreases lung T- and B-cell populations compared with chow feeding. In experiment 2, CFSE+ T-cell retention is highest on day 1 in lung and LP, and decreases on day 2. Cells are gone by day 7; 98.1% of retained donor lung T cells migrate to recipient lungs and 1.9% to the intestine on day 1. Similar results are seen in experiment 3 after transfer of cells to PN-fed recipients. CONCLUSIONS: PN reduces pulmonary lymphocyte populations consistent with impaired respiratory immunity. Transferred lung T cells preferentially localize to recipient lungs rather than intestine with maximal accumulation at 24 hours. Limited cross-talk of transferred lung T cells to the intestine indicates that mucosal lymphocyte traffic might be programmed to localize to specific effector sites. PMID- 19556611 TI - Population patch-clamp electrophysiology analysis of recombinant GABAA alpha1beta3gamma2 channels expressed in HEK-293 cells. AB - Gamma-amino butyric acid (GABA)-activated Cl- channels are critical mediators of inhibitory postsynaptic potentials in the CNS. To date, rational design efforts to identify potent and selective GABA(A) subtype ligands have been hampered by the absence of suitable high-throughput screening approaches. The authors describe 384-well population patch-clamp (PPC) planar array electrophysiology methods for the study of GABA(A) receptor pharmacology. In HEK293 cells stably expressing human alpha1beta3gamma2 GABA(A) channels, GABA evoked outward currents at 0 mV of 1.05 +/- 0.08 nA, measured 8 s post GABA addition. The I(GABA) was linear and reversed close to the theoretical E(Cl) (-56 mV). Concentration response curve analysis yielded a mean pEC(50) value of 5.4 and Hill slope of 1.5, and for a series of agonists, the rank order of potency was muscimol > GABA > isoguvacine. A range of known positive modulators, including diazepam and pentobarbital, produced concentration-dependent augmentation of the GABA EC( 20) response (1 microM). The competitive antagonists bicuculline and gabazine produced concentration-dependent, parallel, rightward displacement of GABA curves with pA(2) and slope values of 5.7 and 1.0 and 6.7 and 1.0, respectively. In contrast, picrotoxin (0.2-150 microM) depressed the maximal GABA response, implying a non-competitive antagonism. Overall, the pharmacology of human alpha1beta3gamma2 GABA(A) determined by PPC was highly similar to that obtained by conventional patch-clamp methods. In small-scale single-shot screens, Z' values of >0.5 were obtained in agonist, modulator, and antagonist formats with hit rates of 0% to 3%. The authors conclude that despite the inability of the method to resolve the peak agonist responses, PPC can rapidly and usefully quantify pharmacology for the alpha1beta3gamma2 GABA(A) isoform. These data suggest that PPC may be a valuable approach for a focused set and secondary screening of GABA(A) receptors and other slow ligand-gated ion channels. PMID- 19556612 TI - Identification and characterization of novel tissue-nonspecific alkaline phosphatase inhibitors with diverse modes of action. AB - Tissue-nonspecific alkaline phosphatase (TNAP) is a ubiquitous enzyme expressed at high levels in bone, liver, and kidney. It appears involved in dephosphorylation of numerous phosphate monoesters, but only 2 of them, pyrophosphate and pyridoxal phosphate, have yet been unequivocally documented. Discovery and characterization of other substrates could be considerably facilitated if specific and potent modulators of TNAP activity with various modes of action were available. Here, the authors describe in detail a high-throughput screening campaign to identify inhibitors of TNAP, performed within the Molecular Library Screening Center Network (MLSCN). A novel homogeneous luminescent TNAP assay was developed and optimized with respect to the enzyme and substrate concentrations, enabling identification of a large number of compounds overlooked by a conventional colorimetric assay. Several new chemical series were identified from screening the Molecular Libraries Small Molecule Repository (MLSMR) collection and demonstrated to have diverse selectivity and mode of inhibition profiles. The nanomolar potency of some of these scaffolds surpasses currently known inhibitors. This article provides an example of a success where the Roadmap Initiative collaborative model, sponsored by the National Institutes of Health, brought together a deep knowledge of target biology from a principal investigator's laboratory, a well-designed compound collection from the MLSMR, and an industrial-level screening facility and staff at the MLSCN center to identify pharmacologically active compounds, with outstanding selectivity data from a panel of more than 200 publicly accessible assays, through a high throughput screen. PMID- 19556614 TI - Do smoking cessation programmes influence geographical inequalities in health? An evaluation of the impact of the PEGS programme in Christchurch, New Zealand. AB - OBJECTIVE: To identify the impact of a smoking cessation programme on area-based social and ethnic inequalities in smoking rates through social and ethnic differences in enrolment and quitting. METHODS: Analysis of records of 11 325 patients who enrolled in an innovative smoking cessation programme in Christchurch, New Zealand between 2001 and 2006. We compare enrolment, follow-up, quitting and impact on population smoking rates in the most and least deprived neighbourhoods and the neighbourhoods with the lowest and highest proportions of Maori. RESULTS: Enrolment as a proportion of the population was higher from the most deprived areas but as a proportion of neighbourhood smokers, it was lower. Enrolees from the least deprived quintile were 40% more likely to quit than those from the most deprived quintile. Smoking rates were 2.84 (2.75 to 2.93) times higher in the most deprived neighbourhoods. If the programme had not been available we estimate that this differential would have reduced to 2.81 (2.72 to 2.90). In neighbourhoods with the highest proportion of Maori, smoking rates were 2.33 (2.26 to 2.41) times higher and we estimate that without the programme smoking rates would be 2.30 (2.23 to 2.37) times higher. CONCLUSIONS: Although enrolees were drawn from a wide variety of backgrounds, those most likely to quit tended to reside in affluent areas or areas with a low proportion of Maori. There was no evidence that this smoking cessation programme increased or decreased inequalities within the Christchurch population. For smoking cessation programmes to have an impact on health inequalities more effort is required in targeting hard-to-reach groups and in encouraging them to quit. PMID- 19556613 TI - Enterohemorrhagic Escherichia coli suppresses inflammatory response to cytokines and its own toxin. AB - Infection with the enteric pathogen enterohemorrhagic Escherichia coli (EHEC) causes a variety of symptoms ranging from nonbloody diarrhea to more severe sequelae including hemorrhagic colitis, altered sensorium and seizures, and even life-threatening complications, such as hemolytic uremic syndrome and thrombotic thrombocytopenic purpura. The more severe consequences of EHEC infection are attributable to the production of Shiga toxin (Stx) and its subsequent effects on the vasculature, which expresses high levels of the Stx receptor, Gb3. Interestingly, the intestinal epithelium does not express Gb3. Despite the lack of Gb3 receptor expression, intestinal epithelial cells translocate Stx. The effect of Stx on intestinal epithelial cells is controversial with some studies demonstrating induction of inflammation and others not. This may be difficult to resolve because EHEC expresses both proinflammatory molecules, such as flagellin, and factor(s) that dampen the inflammatory response of epithelial cells. The goal of our study was to define the effect of Stx on the inflammatory response of intestinal epithelial cells and to determine whether infection by EHEC modulates this response. Here we show that Stx is a potent inducer of the inflammatory response in intestinal epithelial cells and confirm that EHEC attenuates the induction of IL-8 by host-derived proinflammatory cytokines. More importantly, however, we show that infection with EHEC attenuates the inflammatory response by intestinal epithelial cells to its own toxin. We speculate that the ability of EHEC to dampen epithelial cell inflammatory responses to Stx and cytokines facilitates intestinal colonization. PMID- 19556616 TI - Prevalence of sexually transmitted disease syndromes in tribal population of central India. AB - BACKGROUND: A syndromic approach has been advocated for identification and management of sexually transmitted diseases (STDs) in countries where diagnostic laboratory tests are not consistently available. A community-based cross sectional study was conducted to discover the prevalence of STD syndromes in tribal population of central India. METHODS: All married men and women in the age group of 15-49 years from selected villages were enumerated by house-to-house visit. Individuals were interviewed using pre-coded, pre-tested questionnaires about STD syndromes of urethral discharge, vaginal discharge, dysurea, genital ulcer, inguinal swelling, scrotal swelling and lower abdominal pain. RESULTS: Of the 2568 individuals interviewed, 326 (12.7%) had at least one STD syndrome. The prevalence was almost double in women (17.6%) than in men (8.4%). The highest prevalence (16.2%) was observed in the age group 30-34 years followed by 35-39 years (14.7%). The commonest syndrome in women was vaginal discharge (16.0%) while in men the commonest syndrome was dysurea (1.8%). CONCLUSION: The low level of STD syndromes among tribal populations offers an opportunity to prevent a potential epidemic in this disadvantaged community. As no baseline data are available, the findings form the basis for future work in this area. PMID- 19556615 TI - Enacting tobacco taxes by direct popular vote in the United States: lessons from 20 years of experience. AB - BACKGROUND: Tobacco tax increases reduce tobacco use, can provide funds for tobacco prevention and enjoy broad public support. Because of tobacco industry influence in legislatures, US public health advocates have shifted the venue for tobacco tax policymaking to direct popular vote 22 times since 1988. METHODS: We combined case studies of individual state campaigns with tobacco industry documents to identify strategies related to outcome. RESULTS: The tobacco industry developed a voter segmentation model to determine which tobacco tax increases it could defeat. Two industry arguments arising from this model often were raised in losing campaigns-the tax increase did not dedicate enough to tobacco control and hospitals and health maintenance organisations would profit. The industry effectively influenced early voters. Success was associated with building a strong base of public support before the campaign, dedicating sufficient funds to tobacco control, avoiding proposals largely devoted to financing hospitals and other medical service providers, effectively engaging grassroots and framing the campaign with clear justifications for cigarette tax increases. CONCLUSIONS: Tobacco tax ballot measures commonly allocated substantial funds to medical services; tobacco companies are becoming more successful in making this use of funds an issue. Proponents' campaigns should be timed to account for the trend to voting well before election day. Ballot measures to increase tobacco taxes with a substantial fraction of the money devoted to tobacco control activities will probably fare better than ones that give priority to funding medical services. PMID- 19556617 TI - A common UCP2 polymorphism predisposes to stress hyperglycaemia in severe sepsis. AB - BACKGROUND: Insulin resistance and hyperglycaemia are common in severe sepsis. Mitochondrial uncoupling protein 2 (UCP2) plays a role in insulin release and sensitivity. OBJECTIVES: To determine if a common, functional polymorphism in the UCP2 gene promoter region (the -866 G/A polymorphism) contributes to the risk of hyperglycaemia in severe sepsis. RESULTS: In the prospective group 120 non diabetic patients who were carriers of the G allele had significantly higher maximum blood glucose recordings than non-carriers (mean (SD) AA 8.5 (2.2) mmol/l; GA 8.5 (2.4) mmol/l; GG 10.1 (3.1) mmol/l; p = 0.0042) and required significantly more insulin to maintain target blood glucose (p = 0.0007). In the retrospective study 103 non-diabetic patients showed a similar relationship between maximum glucose and UCP genotype (AA 6.8 (2.3) mmol/l; GA 7.8 (2.2) mmol/l; GG 9.2 (2.9) mmol/l; p = 0.0078). CONCLUSIONS: A common, functional polymorphism in the promoter region of the UCP2 gene is associated with hyperglycaemia and insulin resistance in severe sepsis. This has implications for our understanding of the genetic pathophysiology of sepsis and is of use in the stratification of patients for more intensive management. PMID- 19556618 TI - High frequency of de novo mutations in Li-Fraumeni syndrome. AB - BACKGROUND: Li-Fraumeni syndrome is an autosomal dominant cancer predisposition syndrome caused by germline mutations in the TP53 gene. The frequency of germline de novo TP53 mutations is largely unknown; few unequivocal de novo mutations have been reported. METHODS AND RESULTS: Of 341 patients with early onset cancer sent for clinical testing to a national reference laboratory, 75 patients had TP53 germline mutations. Five (7%) de novo mutations were identified, as well as an additional 10 TP53 germline mutations likely to be de novo by family history. The frequency of de novo TP53 mutations in this patient sample is at least 7% and may be as high as 20%. CONCLUSIONS: The possibility that de novo germline TP53 mutations are relatively common has implications for testing and the identification of potential Li-Fraumeni syndrome in patients with little or no family history of cancer. PMID- 19556619 TI - Contribution of RET, NTRK3 and EDN3 to the expression of Hirschsprung disease in a multiplex family. AB - BACKGROUND: Hirschsprung disease (HSCR) is a developmental disorder caused by a defect in the neural crest neuroblast migration process. It is considered to be a paradigm of complex disorders, with many loci contributing to manifestation of the disease. Although HSCR commonly appears as a sporadic trait, approximately 20% of HSCR cases are familial, with complex patterns of inheritance. METHOD: A multiplex HSCR family with an additive model of inheritance, in which the contribution of three genes (RET, NTRK3, EDN3) leads to the HSCR phenotype is reported. RESULTS AND DISCUSSION: The findings suggest that both RET and NTRK3 mutations acting together are necessary and sufficient for the appearance of the disease, and that the EDN3 mutation is acting as a phenotype-modifier factor in the context of this family, as two different HSCR phenotypes are seen among the affected members: a short segment form, and a total colonic aganglionosis. The results therefore support the complex additive model of inheritance previously proposed for Hirschsprung disease. PMID- 19556620 TI - Oxygen uptake kinetics in children and adolescents: a review. AB - The pulmonary oxygen uptake (pVO2) kinetic response at the onset of exercise provides a noninvasive window into the metabolic activity of the muscle and a valuable means of increasing our understanding of developmental muscle metabolism. However, to date only limited research has been devoted to investigating the pVO2 kinetic response during exercise in children and adolescents. From the rigorous studies that have been conducted, both age- and sex-related differences have been identified. Specifically, children display a faster exponential rise in the phase II pVO2 kinetics, which are purported to reflect the rise in muscle O2 consumption, during moderate, heavy and very heavy intensity exercise compared with adults. Furthermore, for heavy and very heavy exercise, the O2 cost of exercise is higher for the exponential phase and the magnitude of the pVO2 slow component is smaller in young children. Sex-related differences have been identified during heavy, but not moderate exercise, with prepubertal boys displaying a faster exponential phase II pVO2 kinetic response and a smaller pVO2 slow component compared with prepubertal girls. The mechanisms underlying these differences are currently poorly understood, and form the basis for future research in this area. However, it is hypothesized that an age-related modulation of the muscle phosphate feedback controllers to signal an increased rate of oxidative phosphorylation and/or altered muscle fiber type recruitment strategies have the potential to play an important role. Overall, the data support the view that at the onset of exercise children have an enhanced potential for oxidative metabolism in the myocyte compared with adults. PMID- 19556621 TI - Effects of group and individual-based step goals on children's physical activity levels in school. AB - This study examined the effect of a 6-week school-based pedometer intervention for children. It compared the number of step counts between group- and individual based step goal conditions over time, and compared the number of goal attainments between the two step goal conditions by physical activity levels. Ninety-nine 4th grade students' (50 boys, 49 girls) data were analyzed. Overall step counts continued to increase over time, increasing about 19% from baseline. Different step goal conditions produced similar effects on children's physical activity levels across all time points. The number of goal attainments was higher for low active children with individual-based step goals than those with group-based step goals. Using pedometers in school is promising for enhancing physical activity in children. PMID- 19556622 TI - Effects of exercise mode on the oxygen uptake kinetic response to severe intensity exercise in prepubertal children. AB - The objective of this study was to verify the effect of the exercise mode on slow component of VO2 (VO2SC) in children aged 11-12 years during severe-intensity exercise. After determination of the lactate threshold (LT) and peak VO2 (VO2peak) in both cycling (CE) and running exercise (TR), fourteen active boys completed a series of "square-wave" transitions of 6-min duration at 75 delta [75% delta = LT + 0.75 3 (VO2peak--LT)] to determine the VO2 kinetics. The VO2SC was significantly higher in CE (180.5 +/- 155.8 ml x min-1) than in TR (113.0 +/- 84.2 ml x min-1). We can conclude that, although a VO2SC does indeed develop during TR in children, its magnitude is considerably lower than in CE during severe-intensity exercise. PMID- 19556623 TI - Modifying middle school physical education: piloting strategies to increase physical activity. AB - Two pilot studies were conducted to examine whether 6th grade students can achieve moderate to vigorous physical activity (MVPA) from 1) activity-based physical education (AB-PE) with 585 participants and 2) a curricular-based (CB PE) program with 1,544 participants and randomly sampled heart rates during lessons. AB-PE participants spent between 54-66% with a heart rate >140 bpm. CB PE participants spent between 49-58% with a heart rate >140 bpm. Girls' mean heart rate was 3.7 bpm lower than the boys. PE can be readily modified so that students spend more than 50% of time in MVPA. PMID- 19556624 TI - Physical fitness and developmental coordination disorder in Greek children. AB - We investigated whether children with suspected Developmental Coordination Disorder (DCD+) demonstrate different physical fitness levels compared with their normal peers (DCD-). Randomly recruited Greek children (n = 177) were assessed for body mass index (BMI), flexibility (SR), vertical jump (VJ), hand strength (HS), 40m dash, aerobic power, and motor proficiency. ANCOVA revealed a motor proficiency (i.e., DCD group) effect for BMI (p < .01), VJ (p < .01), and 40m speed (p < .01), with DCD+ children demonstrating lower values than DCD-. Differences between DCD+ and DCD- were also obtained in log-transformed HS (p < .01). These findings suggest that intervention strategies for managing DCD should also aim at physical fitness increases. PMID- 19556626 TI - Personal characteristics and demographic factors associated with objectively measured physical activity in children attending preschool. AB - Independent associations between personal- and demographic characteristics and physical activity in 3-6 year old children attending preschool were identified in this study. Boys spent a larger proportion of the time on moderate-and-vigorous physical activity (MVPA; p < .001) and had a higher total physical activity level compared with girls (p < .001). The 3-4 year old children spent less time on MVPA and had a lower total physical activity level compared with both 4-5 (p < .01) and 5-6 year old children (p < .001). The individual preschool, gender and age of preschool children were strong predictors of physical activity (R2-total model=(0.36-0.39)) during preschool attendance. PMID- 19556627 TI - Allometric associations between body size, shape, and physical performance of Greek children. AB - We adopted allometric models to identify the most appropriate body size/shape characteristics associated with physical performance activities of Greek school children. Children underwent assessments for aerobic and anaerobic fitness, flexibility and hand-grip strength. Results suggest that the inverse Ponderal index and not BMI is the most appropriate body-shape indicator associated with running and jumping activities. Height was negatively associated with flexibility, but both height and weight were positively associated with hand-grip strength. In conclusion, allometric models provide a valuable insight into the most appropriate body size and shape characteristics associated with children's physical performances and at the same time ensure valid inference when investigating group/population differences (e.g., between gender and maturation status). PMID- 19556625 TI - Factors related to objectively measured physical activity in preschool children. AB - This study examined correlates of objectively measured physical activity (PA) in a diverse sample of preschool children (age 3-5 years; n = 331). Accelerometer min x hr-1 of moderate-to-vigorous physical activity (MVPA) and nonsedentary activity (NSA) were the outcome measures. Correlations among potential correlates and PA ranged from r= -0.12-0.26. Correlates in the final MVPA model were age, race, sex, BMI Z score, and parent perception of athletic competence, explaining 37% of the variance. The NSA model included the latter two variables, explaining 35% of the variance. Demographic factors were correlates of PA; parent perceptions of children's. PMID- 19556628 TI - Association of perceived environmental characteristics and participation in organized and non-organized physical activities of adolescents. AB - Specific behavior context such as type of PA (organized vs. nonorganized) might be associated with different environmental correlates. The main goal of this cross-sectional survey was to examine perceived environmental associations with type of adolescents' physical activity (PA) choices (organized and nonorganized). A sample of this study comprised 425 girls with mean age of 14.5 years-old. Environmental variables and PA were assessed by questionnaire, which allowed to define the type (organized or nonorganized) of PA. No associations were found between environmental perceptions and the participation in organized activities. However, different dimensions of environmental variables such as accessibilities to facilities (p -/+ .05) aesthetics (p -/+ .05) and social environment (p -/+ .05) were associated to girls' PA participation in nonorganized activities (NOPA). Our findings suggested that some environmental characteristics might play an important role in girls' NOPA participation. PMID- 19556629 TI - [Comparative characteristics of various anesthetic methods during coronary artery bypass grafting CABG]. AB - During the last 10 years increase in the number of surgical centers developing the practice of off-pump heart surgeries has been noticed. Our medical centre, where 140 CABG surgeries have been performed, is one of the like. Such surgeries demand multicomponent anesthetic manual able to provide adequate depth of anesthesia, hemodynamic stability, prevention of intraoperative myocardial ischemia, providing an early patient activation in a postoperative period. On the assumption stated above we suggest and compare two anesthetic methods - total intravenous anesthesia on the basis of Propofol and Phentanylum and inhalational anesthesia on the basis of Isoflurane for providing CABG Surgery. It has been detected that the anesthetic methods offered by us enable to provide an adequate anesthetic defence while performing CABG Surgeries in case of stable markers of hemodynamics and oxygen transfer functions. Mild hemodynamic disorders have been detected in the operation course first of all at the expense of positional changes and heart immobilization. These disorders have not lead to the deterioration of oxygen transfer functions and have been totally eliminated in the recovery period. Worked-out anesthetic methods on the basis of Midazolam, Propofol, Isoflurane and low doses of Phentanylum (5-7 mcg/kg per hour) give a possibility to decrease the period of artificial lung ventilation, provides an early (less than 6 hours) and safe extubation for 86% of the patients and reduces their stay in a resuscitation unit as well. PMID- 19556630 TI - [The results of the treatment of gunshot wounds by applying laser therapy]. AB - The treatment results of not penetrating in cavities surface gunshot wounds of border regional hospitals are studied. The injured are divided into 3 groups depending on the terms of first surgical processing of wounds. It is mentioned that putting dense sutures is not always justified. Very often inflammatory infiltrate and suppuration is arisen after suturing. Inflammatory process develops into tumoral and cicatricial disorders which are removed by surgical intervention. The gunshot wound processing with helium-neon red ray made favorable conditions for early closing of wound surface by means of saturation with bringing the edges of the wound closer and preventing the development of severe suppuration complications as well as reduced the treatment terms promoting good functional and cosmetic results. PMID- 19556631 TI - Otosclerosis surgery: approaches, profits and complications. AB - A systematic analysis of stapedoplasty output in otosclerosis cases was carried out. The operations were done during the period of 2005-2008 years at the Department of Otorhinolaryngology of the Tbilisi State Medical University. From the overall number of 107 patients, 78 were females and 29 males, 72.9% and 27.1%, respectively. The ages ranged from 16 to 57 years. The mean age was 35 years. The conductive and mixed forms of hearing losses were diagnosed in 70 and 37 cases, 65.4% and 34.6%, respectively. Right ear was operated in 46 patients, left ear in 57, and both ears in four, 43.3%, 53.0%, and 3.7%, respectively. 111 ears have been cured thus in sum. Intumescences of external ear tube were observed during the operation in 22 patients, 20.6%. In 7, 6.5%, the facial nerve was located downward. Particularly abnormal placement of the facial nerve was the case in two patients, 1.9%. The endaural approach has been proved to own advantages over the transmeatal one. The data generally confirmed that stapedotomy, as compared to stapedectomy, is a better choice for the surgery output. In beneficial cases the air/bone gaps after the operation closed totally or nearly totally. Such a proper outcome was reached in 93 out of 111 ears operated, 83.8%. In most of remainder ears the gaps after the operation narrowed significantly but far not completely. The definite surgery failure happened in one case only, 0.9%. To balance the preserved middle-ear problems in non-perfect surgery cases, the hearing aids of bone-conduction types have been recommended. In mixed otosclerosis cases, conversely, the aids of air-conduction types were advised to overcome the coexisted inner-ear pathologies. PMID- 19556632 TI - [Venous thrombotic complications in pregnancy: way of prophylaxis and therapy]. AB - The aim of this investigation was to study various aspects of hereditary thromboses in pregnant women with gene anomalies (considering regulatory role of genes in hemostasis in norm and in hereditary diseases that disturb blood coagulation). In comparative aspect frequency of occurrence of hereditary thrombogenic anomalies (F-V-L, F-II-20210, MTHFR) in healthy women during and after pregnancy, in pregnant women with thrombosis and in those who had thrombosis in previous pregnancy was studied. Patients were divided into four groups. The first group included 103 pregnant women with signs of a thrombosis, thrombophlebitis, varicose veins expansion of lower limbs etc. in anamnesis. The second group was composed of 110 women with surface varicose veins expansion of lower limbs during the pregnancy. The third group consisted of 90 not pregnant women with varicose veins expansion. The fourth group (control) consisted of 76 pregnant women without thrombogenic anomalies. All the examinations were performed by a standard technique (three-dimensional color Doppler sonography. Pregnant women were treated with direct action anticoagulants in case of thrombotic and prethrombotic conditions. A policy for individualized treatment is needed in pregnant women with hereditary thrombogenic anomalies to determine the doses and the period of anticoagulant therapy. PMID- 19556633 TI - [Root canal treatment at pulpitis, periodontitis using ultrasound]. AB - The aim of our research was the clinical estimation of ultrasound efficiency during root canal treatment at pulpitis, periodontitis. 446 root canals of 164 single and multi-rooted teeth were used for this study. Teeth were treated and monitored for 3 months, 6 months, 9 months, 1 year, 1,5 year, 2 year. All teeth with periapical destructive changes had periapical destructive lesions in x-ray imaging. Patients were divided into two groups. Endodontic treatment of first group combined mechanical and chemical processing with ultrasound divise Pieson Master-400, Endosonore profiles, sodium hypochlorite (NAOCI) +canal lubricant (EDTA). In a second group we used the traditional layout of treatment pulpitis end periodontitis. According to our results, in a first group the X-ray improvement was - after 3 months in I group in 53 (64,6+/-0,7%) patients, in the II group - 44 (53,65+/-2,3%); after 6 months in the I group - 59 (71,9+/-2,2%), in the II group - 56 (68,29+/-0,54%); after 9 months in the I group - 62 (75,6+/ 0,56%), in the II group - 56 (68,29+/-0,54%); after 1 year in the I group - 70 (85,36+/-0,18%), in the II group - 63 (76,83+/-0,53%); after 1,5 year in the I group - 76 (92,68+/-2,8%), in the II group - 65 (79,26+/-0,5%); after 2 years in the I group - 77 (93,9+/-2,7%), in the II group - 65 (79,27+/-2,11%). We can conclude, that the ultrasound preparation of the root canal leads us to the considerable lowering of complication of endodontic treatment, though doesn't guarantee their complete absence. In the end we can resume that the usage of ultrasound preparation during endodontic treatment considerably exceed the effectiveness of chemical irrigation of root canal. That is because of its bactericide effect, opening and cleaning root canal system, creation favorable conditions for hermetic filling of root canal after removal of smear line. PMID- 19556634 TI - [Comparative estimation of laser devices in complex treatment of oral cavity mucous membrane diseases]. AB - The aim of the study was to compare laser devices "Optodan" and portative laser "Baure" in complex treatment of oral cavity mucous membrane diseases. We studied 90 patients: 54 female (60,0+/-0,77%) and 36 (30,0+/-0,7%) men from 18 to 45 years old. All patients had different forms of oral mucosal diseases: Stomatitis aphtosa chronica recidiva had 36 patients (28,9+/-0,9%); Erythema exudativum multiforme - 10 patients (6,7+/-1,5%), Candidosis angulitis - 26 patients (17,8+/ 1,46%), Lichen ruber planus ulceroza - 4 patients (4,44+/-1,03%), Trauma mechanicum acutium - 12 patients (11,06+/-2,8%), Stomatitis herpetica - 28 patients (31,1+/-1,48%). All the patients were practically healthy and had no contraindications for physiotherapy. Before treatment to all patients had been recommended means of individual hygiene and several recommendations. The patients were divided into two groups. Every group consisted of 45 patients. 13 patients with Stomatitis aphtosa chronica recidiva, 14 - Stomatitis herpetica, 3 - Erythema exudativum multiforme, 8 - Candidosis angulitis, 2 - Lichen ruber planus ulceroza, 5 - Trauma mechanicum acutium. All the patients underwent symptomatic treatment according to the form and gravity of disease. Patients in a first group in addition had laserotherapy with a laser device "Optodan". Patients in a second group underwent laserotherapy with a portative device (Baurer). The positive results were achieved in both groups, though the first group patients mentioned considerable reduce of pain right after procedures. Our research proved the high effectiveness of laserotherapy in complex treatment of oral cavity mucous membrane diseases. Treatment with device "Optodan" is the most preferable. PMID- 19556635 TI - [Densitometric investigation of the dynamics of bone defects regeneration in surgical treatment of chronic periodontitis]. AB - Apicotomy of 105 teeth of 78 patients with chronic periodontitis has been performed. The periapical defect were filled with two kinds of osteoplastic matter: the first group was treated with demineralised bone matrix of newborn pigs (DBMNP), the second group with artificial hydroxyapatite, while both matters were enriched with platelet rich plazma (PRP).As a third, control group, cases. The defects of which were not filled with osteoplastic matter, were investigated. The dynamics of the regeneration of bone defects was studied based on the data of densitometric investigation. The results of objective observations have revealed high efficiency of surgical treatment of chronic periodontitis with the filling with DBMNP in combination with PRP. PMID- 19556636 TI - Overview of referrals to mental health services in Georgia. AB - The prevalence of mental illnesses in society is difficult to measure accurately as current techniques are relatively poor. Mental health professionals argue that data provided by Ministry of Health of Georgia concerning the prevalence of mental illnesses should be treated with great caution as the official rates of mental disorders are likely to be largely underestimated. The most significant reason of this could be focusing on referral to state mental health institution and ignoring the private sector. In order to understand the gap between official data and real needs in mental health care we compare statistical data of state institutions and private practice. The structured questionnaire was used for obtaining referral information from the Asatiani Psychiatric Hospital, Gotsiridze Psycho-Neurological Dispensary and a private psychiatrist. RESULTS: The state institutions with great proportion serve patients with schizophrenia, schizotypal, delusional and organic mental disorders, while patient with mood disorder and neurotic, stress-related and somatoform disorders preferred private psychiatrist. People with different psychiatric problems (e.g. sleep and eating disorders), especially young and elderly patients seldom are seen by psychiatrist. CONCLUSIONS: Hence, the official data concerning the prevalence of mental illnesses in Georgia are highly underestimated and could not reflect real demands. PMID- 19556637 TI - [The comparative study of Linex and Lacto-G in treatment of adult patients with disbacteriosis]. AB - The quantitative and qualitative disturbance of structure of intestine microflore show negative influence of process of adoption and digest of food stufis, causes allergy of organism and has predisposition for development of infection. 100 patients were investigated with chronic disfunction of intestine-50 patients in each group. In all patients were investigated coprogramme before treatment, after 2 weeks and one month. The patients of the first group were treated with Linex, in the second group - with Lacto-G. In both groups before treatment disbacteriosis was revealed. Investigation of feces showed pathological flora in 18% and 22% cases accordingly. Improvement of microflore of intestine in both groups was noticed after treatment. The results of the clinical and laboratory tests showed that one month course of treatment improved clinical and microbiological parameters. PMID- 19556638 TI - [Cryotherapy in treatment of skin demodecosis]. AB - Demodecosis is wide spread chronic skin disorder caused by the mite - Demodex folliculorum. It frequently involves the mid facial region: nasolabial folder, the nose, the chin and eyes. The treatment depends on the severity of the inflammatory skin disorder and includes complex of oral and topical drug treatment. The aim of the study was to determine the efficiency of criotherapy in complex treatment of demodecosis. We have observed 38 patients of both sexes aged 19-54. The patients were divided into two groups - "A" and "B". Patients with erythematosquamose form of the disease were treated with Trichopol peros and oral antibiotics in case of inflammatory lesions. For topical treatment in group "A" Rosamet cream was used. In group "B" Rosamet cream and criotherapy were used. The results of our study have demonstrated effectiveness of criotherapy in combination with Rosamet cream. A significantly faster decrease of inflammatory lesions and reduce the erythema was observed in group "B". Therefore the usage of criotherapy with Rosamet cream is recommended to get quick therapeutic effect. PMID- 19556640 TI - [Comparative study of different diagnostic methods in pulmonary alveolitis]. AB - To evaluate and analyze, clinical and roentgenological manifestations of extrinsic allergic alveolitis (EAA) and idiopathic fibrosing alveolitis (IFA) 89 patients underwent bronchoscopy survey and functional test of lungs. Average age of patients was 38,3+/-5,8. Among examined 89 patients 31 (34,8%) patients were poultry farmers, 30 (33,7%) - millers, and 28 (31,5%) - tobacco-growers. EAA was found among 22 poultry farmers, 19 tobacco-growers, and 19 millers. IFA was found among 11 millers, 9 tobacco-growers and 9 poultry farmers. Acute respiratory disease (ARD) was found among 58 patients; 38 patients suffer from lung diseases. Control group consisted of 20 healthy people. Along with general blood analysis, all patients underwent - roentgenological analysis of thorax in two shifts. Recent studies show that CT lung screening is more sensitive than standard lung screening methods in detecting lung disease. Comparative analysis allowed concluding that Real-Time CT method is the most effective. CT lung screening is more sensitive than standard lung screening methods in detecting lung diseases. PMID- 19556639 TI - [The complex approach to treatment of pain syndrome of cervical vertebral osteochondrosis]. AB - The article deals with an investigation of therapeutic efficiency combined with applications of biological preparations and special medical-improving gymnastic exercises at treatment of a vertebral cervical osteochondrosis on the one hand, traditional means on the other hand. 76 patients aged 31-62 have taken part in the given research. The basic group was made by 47 patients and the control group - by 29. The patients of the basic group applied a special complex of medical improving gymnastic exercises, and also paravertebral injections of biological preparations Traumeel S, Neuralgo-Rheum-Injeel. The patients of the control group received traditional therapy of a vertebral osteochondrosis. Injections of Voltaren (Diclofenac) intramuscularly, MIG400 (Ibuprophen) tablets, phonophorez with Indometacin ointment, a traditional complex of physiotherapy exercises. Duration of the treatment in all groups has made 28 days. Obtained data testify that by the end of treatment full knocking over of a pain syndrome in the basic group is noted at 71,2 % of patients, in control group only in 41,4 % of patients the pains have completely disappeared in a cervical spine. Thus, therapy by biological preparations in a combination to a complex of special medical improving gymnastic exercises allows to reduce terms of knocking over of sharp clinical displays of an osteochondrosis of a cervical spine, to raise efficiency of restoration of sensitive, vascular, muscular-tonic frustration, and also allows to avoid neuro-surgical intervention at patients with hernias interspine disks. PMID- 19556641 TI - [Comparative estimation of cytokine profile among patients with exogenous allergic and idiopathic fibrosing alveolitis]. AB - To study some indicators of cytokine status among the patients with pneumonia alveolitis 89 patients have been examined. 60 of those patients suffer from exogenous allergic alveolitis (EAA) and - 60 from idiopathic fibrosing alveolitis (IFA). The average age is 36.4+/-3.0. The patients were devided into three groups: I group - 21 (23.6%) patients with acute disease, II group - 25 (28.1%) patients with subacute disease, III group - 43 (48.3%) patients with chronic disease. The level of IL-1beta and IL-8 in blood serum and bronchoalveolar wash out (BWO) measured by IFA test system. It is identified that the contents of IL 1beta in blood serum of EAA patients exceeded on 7.2%, the contents of IL-8 exceeded on 14.6% among IFA patients - reliable rise IL-1beta on 35.4% (<0.05) and IL-8 on 31.6% (<0.05). The level of proinflammatory cytokines (IL-1beta and IL-8) in peripheric blood corresponded with the level of disease. Along with progressing of disease the contents of cytokine in respiratory space, too high, increased among 21 patients with acute disease, in 13 cases (61.9%) BWO was high in IL-1beta and in 80.8% cases of IL-8. in II group among 25 patients with subacute disease high level of IL-1beta marked in 56.0%, Il-8 in 84% cases; patients in III group high concentration of IL-1beta registered in 51.2%; Il-8 in 81.4% of patients. PMID- 19556642 TI - [Megaloblastic-vitamin B12 deficiency anemia in childhood]. AB - Megaloblastic anemias are basically caused by vitamin B(12) and/or folic acid deficiency. Childhood vitamin B(12) deficiency is extremely rare. There are congenital and acquired forms of vitamin B(12)-deficiency anemias. The article captures findings of 10 year observation of 3 patients with Imerslund-Grasbeck Syndrome (congenital chronic megaloblastic anemia with proteinuria), in which the diagnosis was established by us in early childhood and due to correct treatment and prevention complete clinical-laboratory remission is kept so far. We have also observed rare case of acquired megaloblastic anemia - 14 years old vegetarian patient, who was diagnosed with vitamin B(12)-deficiency anemia based on history, clinical and para-clinical data. It was caused by strict vegetarianism of the patient. Therefore first of all the diet was corrected. In 5 days of specific treatment with vitamin B(12) "reticulocyte crisis" was manifested (proving the correctness of diagnosis and treatment) and complete clinical-hematological remission was achieved in 2 weeks. The given cases are interesting as megaloblastic anemias in childhood are both rare and difficult to diagnose. In such cases timely diagnosis, treatment and prevention tactics should be based on cause-and-effect relation of disease. PMID- 19556643 TI - [Collonization of intestinal tract of leukaemia patients with Candida and development of systemic candidemia]. AB - The aim of the research was to study the peculiarities of colonization of intestinal tract by Candida - in children with leukemia. It was found that in 32 out of 54 patients with leukaemia the quantity of intestinal KOE Candida in 1 g feces was correlated with the incidence of candidemia. The quantity of intestinal KOE/g Candida was higher in patients receiving antileukemic chemotherapy and antibacterial antibiotics than in normal subjects in patients with The quantity of Candida KOE/g feces was >10(5) cells per 1g of substrate. In control group this figure was within 10(3)-10(4). In all patients candidemia was revealed. In 8 patients who were not treated with antibiotics the quantity of Candida in feces was present to a less degree (<0.05). In the majority of cases (70%) in patients with leukaemia C.albicans was found, in 15% of cases - C. tropicalis. Along side with candidemia candidiazes of respiratory and urinary systems was revealed in children with leukaemia. PMID- 19556644 TI - [Influence of different type beta-adrenoblockers on functional state of liver at paracetamol induced toxic hepatitis in experiment]. AB - The aim of the present work was comparative analyze of different type beta adrenoblockers (nonselective beta-AB-Propranolol; cardio selective beta1-AB Bisoprolol; nonselective beta-AB with additional vasodilatory effects - carvedilol) on functional state of liver at Paracetamol induced toxic hepatitis in experiment. Experiments were carried out on 42 laboratory white rats with the body mass 180-200 g. Model of acute toxic hepatitis was created using paracetamol (peroral administration with the dose of 1000 mg/kg, once a day). Beta adrenoblockers (beta-AB) were used with following dozes: propranolol -1,4 mg/kg; bisoprolol - 0,7 mg/kg and carvedilol - 0,9 mg/kg; beta-AB were used during 9 days (perorally), immediately after paracetamol administration. Each group consisted of 7 rats. Functional state of the liver was estimated according to blood serum concentration of aspartat-transferase (AST) and alanin-transferase (ALT) using routine method of measurement on 5th and 9th days after administration of paracetamol. Results of experiments have shown that single administration of paracetamol with the dose of 1000 mg/kg impaired functional state of the liver that was confirmed by significantly increased activation of transaminases. It was stated that non-selective beta-AB Propranolol worsens-, cardio selective beta1-AB Bisoprolol - has no effect, while nonselective beta-AB carvedilol, with additional vasodilatory and antioxidant effects - slightly improves functional state of the liver at acute toxic hepatitis caused by paracetamol. Has been suggested that hepatotoxicity is not characteristic for all types of beta-AB. According to the obtained results, it has been recommended to estimate functional tests of the liver before propranolol administration, and carry out monitoring of the liver's functional state at treatment course, especially in individuals suffering with liver malfunction. It is important to investigate detailed mechanisms of hepatotoxicity of beta-AB also. PMID- 19556645 TI - [The perspectives of using lectins in immunodiagnostic and gene therapy]. AB - In this article review diagnostic value of protein group-lectins for practical oncology are presented. It is indicated, that with some carbohydrate structures lectins revealed the possibility of reversible and selective action and with earlier elaborated methods, which are based on identification of antigen determinants with antibodies, they can significantly increase of tumour diagnostic sensitivity and specificity. It is consider the mechanisms of relationship between so-called mannan-binding lectins and mannans or carbohydrate structures, which are formed from the mannose residue. The presense of such mannose-enrich structures is connecting with the process of blastomic transformation. It was shown experimentally, that mannan-bindig lectins are interacting with the carbohydrate chains of the markers of blastomic transformation, particularly with the cancer-embryonic antigen. It was established the markedly increase of lectins gene expression in transformated cells. The gene therapy, which comprises the deliver of therapeutic genes to cells and tissues by virus vectors and nonvirus systems is also analysed. The last of them are the most widely using ligands containing oligosaccharides, because on the surface of many animal origin cells were identified lectins protein receptors. It is suggested the perspectives of using oligosaccharide probes, which are the ligands for endogenous receptors-lectins in tumour diagnostic and guiding transport of medicinal products. PMID- 19556646 TI - [Analgetic property of nonspecific thalamic nuclei]. AB - Neruronal activity evoked by pain impulsation in the specific thalamic nuclei and sensorimotor cortex has been recorded in cat. The following data were obtained. Evoked nociceptive neuronal activity was inhibited by stimulation of the nonspecific thalamic nuclei. An antinociceptive impact of the nonspecific thalamic nuclei on the specific thalamic units was fulfilled by both direct and thalamo-cortico-thalamic pathways. Stimulation of the antinociceptive regions of the nonspecific thalamic nuclei abolished evoked nociceptive activity of the cortical units, elicited by both peripheral noxious stimulus and stimulation of the ventro-postero-medial thalamic nuclei. This indicates that the nonspecific thalamic nuclei inhibit the cortical activity on the thalamic, as well as, on the cortical level. PMID- 19556647 TI - Timing of action potentials and input currents in perisomatic and trilaminar interneurons during kainate induced gamma oscillations in mice CA1 hippocampus. AB - Nonaccommodating perisomatic basket cells had been suggested to structure gamma oscillations. Here we used simultaneous whole-cell patch clamp recordings within 150 microm of CA1 stratum pyramidale and extracellular population activity measurements in CA1 stratum radiatum of the mice hippocampal slices to show that though nonaccommodating perisomatic interneurons and trilaminar cells convey information through the same frequency of action potentials during the kainate micropressure ejection induced gamma oscillations, inhibitory inputs received by soma and proximal dendrites of pyramidal cells differ with pi radian in time. Perisomatic interneurons discharge action potentials (APs) at the positive peak of the field gamma waves, whereas trilaminar cells fire at the trough. Timing of excitatory currents in each type of interneurons coincides corresponding APs discharge time during the active network. Perisomatic interneurons receive gamma modulated very high frequency EPSCs, while trilaminar cells receive only gamma frequency excitatory inputs. Inhibitory postsynaptic currents lag the action potentials in both cases with pi/2 intervals. Trilaminar cells APs discharge is firmly phase locked to the trough of the field gamma waves, while nonaccomodating perisomatic interneurons spike firing rotates "counter-clockwise" during the oscillations. Apical dendrite inhibition through trilaminar cells seem to play similarly important role as do nonaccommodating perisomatic cells in the formation of 30-80 Hz oscillations. In this study we showed the posibility of existence of different active events during the gamma oscillations in two strata of CA1 that are produced by the discharge of perisomatic inhibitory interneurons in the stratum pyramidale and trilaminar cells activity in the stratum radiatum after the time intervals. PMID- 19556648 TI - Ultrastructural and molecular-biological features of inflammatory-destructive processes in pathology of parodontal complex in children. AB - In children aged 11-15. under mild and moderate stage parodontitis the ultrastracture and Citokeratines 10/13 and 14 expression in epithelial lining of oral mucosa were analyzed: 1. in gingival epithelia 2. in alveolar processes epithelia. 14 cases without sings of inflammation serve as control tissue. Total number of cases - 33. After informed consent had been obtained, simples of histological tissue specimens were collected on surgical extraction of the tooth. In the control group decision on the tooth extraction was taken for the orthodontic causes. Our data indicate that: 1. Heterogenity is typical to the oral cavity epithelium: a) Ultrastructural signs of keratinization and dissociation, with typical high activity of the terminal differentiation marker cytokeratin 10/13, predominate in the keratinocytes of gingival mucosa. b) Cells with signs of germination activity predominate in the ultrastructure of mucosa alveolar processes. Such cells express cytokeratin 14, typical to nonkeratinized epithelium. 2. Tissue architectonics as well as protein contents of cytoskeleton (judging by cytokeratine expression) are speared in the parodontal pathology in children, however in contrast to alveolar mucosa, damage to the microcirculatory vessels is more pronounced in gingival mucosa. 3. Expression of cytokeratines 10/13 and 14 may indicate the process of lysis and reparation of periodontal ligament. PMID- 19556649 TI - [The age-dependent peculiarities of quolitative analysis of intestinal villi morphogenesis]. AB - The aim of our investigation was to study morphogenesis of microcirculatory bed of small intestine villi in rats. The research was conducted on 48 (male and female) 1 day, 7 day, and 17 days old white rats by using scanning electron microscope and metrical parameters of the villi components. Measurement of the microvessels profile area in the shear plane on different levels of fiber is changed. Before feeding of the animals it is decreasing and after feeding it is increasing. We came to the conclusion, that microvessels are in all parts of villi, especially in new born before feeding and in all ages too. And there is a great tendency of lowering this index in all ages from top to basis (In 1 day animals 4 times; in animals 7 day 3 times; and in 17 day old animals 2 times. PMID- 19556650 TI - Morphological characteristics of tissue damages from electrical contact. AB - The purpose of the study was to reveal the morphological pattern of different tissues mostly vulnerable to electric contact injury from domestic electrical appliances and to determine the possible diagnostic criteria of this damage. The matter is of particular importance as domestic electrical appliances are widely used in torture and other cruel, inhuman and degrading treatment or punishment. The pilot part of the study was experimental and as the material of study have been used adult Wistar white rats. The microscopic study of slides taken from tissues damaged by electricity and dyed by routine Hematoxilin-Eosin reveals general structural changes that does not represent characteristic morphological pattern sufficient for forensic diagnosis of electric injury. The electricity damage of kidney and adrenal glands reflects the morphological pattern of stress that allows complex evaluation of damage but could have only orientating value for the estimation of cause of injury. Due to results obtained from pilot part of the study there is considered in regard to determine the possible diagnostic criteria for electrical injury of tissues the experimental morphological study must continue on skin and myocardial material using routine Hematoxilin-Eosin as well as other classical and modern morphological methods of study. PMID- 19556651 TI - The positive effects of Midazolam on functional activity of white rat brain cells in conditions of halothane anesthesia. AB - For the elimination of postoperative complications, which are evident in infants with congenital palatine and upper lip cleft after halothane anesthesia during standard premedication (Atropine, Dimedrol) the usage of agents of benzodiazepine group with anti-hypoxic effects is advised. The latter may modulate the blocked ion channels by neuromediators, which are activated by GABA and GABA receptor function. The neuro-protective ability of halothane is demonstrated. To reveal the mechanisms of positive effects using benzodiazepine group for premedication we have investigated the effects of midazolam of brain cell functional activity of experimental animals (white rats) in conditions of halothane anesthesia. For the estimation of white rat brain cells functional activity the nuclear transcriptional activity was studied (based on the intensity of [14C]-UTF inclusion), also the glutamic acid decarboxilaze activity (GAD65/67) using immunohistochemistry. It is estimated that halothane inhibits the transcription in rat brain cells. During midazolam premedication the halothane inhibitory effect on RNA synthesis is not revealed. After an hour of pseudo-operation halothane also induces proved decrease of quantity of GAD65/67 positive cells in CA3 hippocampal field. At the same time the quantity of similar cells are increased in CA1 field. The increase of GAD65/67 positive cells in CA1 is more evident during midazolam premedication. Based on the data received we can conclude that the positive effect of midazolam results in increase of GAD65/67 positive cells in CA1 hippocampal field. PMID- 19556652 TI - [Using of chromatographical methods to study stability, pharmacokinetics and metabolism of propofol (review)]. AB - Propofol (somnopol, diprivan) 2,6-diisoprophilphenol is a preparation of short term, intravenous narcosis used for local and general anaesthesia through single injection or permanent infusion with the speed of 4-12 mg/kg in an hour. In this study we tried to summarize the experience of using of chromatographical methods (gas chromatography-GCH0, high performance liquid chromatography (HPLC), thin layer chromatography (TLCH) and combination of mass-spectrometry with GCH (GCH MC) and HPLC (HPL-MC) to study stability, pharmacokinetics and metabolism of propofol. A labour-intensive and important work have been carried out to obtain information about propofol. PMID- 19556653 TI - Double-step procedure screening for modeling reactive (exogenous) depression in rats. AB - Depressive disorders, which show ever increasing prevalence, urgently require relevant medications. In its turn search for the antidepressant drugs requires valid animal models of depression for a drug screening. The goal of the present study was construction of affordable and simple model of reactive depression in rats, which model could be implemented in the antidepressants screening. To this end the two experimental paradigms have been modified and combined in groups of rats, which had no signs of depressive behavior. In the first experiment animals were subjected to the so-called immobilization stress. In the next step the animals, in which immobilization stress has been elicited, were introduced into the modified Forced Swim Test (FST) apparatus with an exercise wheel. Testing procedure in the FST apparatus lasted 10 min, and it was determined that 120 revs/10 min was minimal activity index for non-depressive animals, while lesser number of revolutions pointed at existence of acquired depressive status. Immobilization-induced depressive behavior was stable enough and remained two weeks following the stress. Therefore, our double-step procedure constitutes versatile, reliable, affordable, and humane screening means for modeling reactive (exogenous) depression in rats. PMID- 19556654 TI - [Prospects of the cultivated medical plants in Georgia]. AB - The aim of the research is to give botanic description, to analyze chemical composition and medical properties of plants. The names of plants in Russian are also provided. The main areas of propagation of medicinal plants are identified. The period of collection is recommended. The most important features of raw materials and extracts are represented. The information about the therapeutic effectiveness of drugs is depicted. PMID- 19556655 TI - [Antidiabetic action of extract of Juglans regia L]. AB - The aim of the research was to study the impact of Iuglans Regia L on the pathological processes in alloxan-indeced dibatic animals. The investigation showed that extract from wall nuts leaves decreases the blood sugar level, has a positive impact on lipid metabolism. Antioxidant properties of an extract from leaves of the walnut tree (Juglans regia) are reported. The extract suppresses functional insufficiency of liver links synthethising enzymes, increases the antitoxic action of hepatocytes, improves the functional insufficiency of kidneys. The extract is recommended in prevention of DM and its late complications. PMID- 19556656 TI - Growth curves for school age children and adolescents. PMID- 19556657 TI - Nutrigenetic determinants of neural tube defects in India. AB - JUSTIFICATION: Neural tube defects (NTDs) are one of the commonest birth defects with a high incidence in India. However, few studies have systematically looked into the etio-pathogeneis of NTDs, which mainly includes nutritional deficiencies and genetic predisposition. Efforts are afoot for universal food fortification with folic acid in the hope of preventing NTDs, without factual evidence of folate deficiency in the target population. EVIDENCE ACQUISITION: We conducted a review of Indian literature on NTDs focusing on the role of folate and vitamin B12 nutrition and common genetic polymorphisms in 1-carbon metabolism. We performed a literature search of Medline and Indian Medlars (www. indmed.nic.in) for articles using following search terms: Neural tube defect and India, published up to November 2008, on human subjects. We did not include individual case reports and case series describing surgical and medical management, genetic syndromes where NTD was only one of the features or unusual associations of NTDs with other clinical findings. RESULTS: Absence of a nationally representative large study, lack of interventional studies and methodological differences were conspicuous during this review. Larger studies are, therefore, urgently needed to delineate genenutrient interactions in association with NTDs in India. We urge that caution should be exercised before widespread folic acid fortification of food, without addressing the issue of concurrent B12 deficiency. PMID- 19556658 TI - Crosssectional growth curves for height, weight and body mass index for affluent Indian children, 2007. AB - BACKGROUND: The assessment of growth is crucial in child care and reference data are central to growth monitoring. As the pattern of growth of a population changes with time it is recommended that references be updated regularly. OBJECTIVE: To produce contemporary growth curves for Indian children from 5-18 years for height, weight and BMI. DESIGN: Cross-sectional. SETTING: Multicentric, School based. PARTICIPANTS: 19834 children were measured from 10 affluent schools from five major geographical regions of India. Data were analyzed on 18666 children (10496 boys and 8170 girls) using the LMS method and smoothed percentiles 2007 were produced. RESULTS: Compared to the 1989 data, median height at 18 years was 0.6 cm greater for boys but unchanged for girls, while the 97th height percentile had increased by 1.7 cm for boys and 2 cm for girls. Boys and girls were heavier and taller at almost all ages. The study also showed that boys and girls were taller at a younger age. CONCLUSIONS: Contemporary cross sectional reference percentile curves for height, weight and body mass index for the assessment of physical growth of present day Indian children are presented. PMID- 19556659 TI - Antibiotic associated diarrhea in children. AB - CONTEXT: Keeping in view the recent flooding of the Indian market with antibiotic and probiotic combinations, we decided to look at the prevalence of antibiotic associated diarrhea (AAD) and Clostridium difficile infection (CDI) in children and reviewed evidence available for use of probiotics in the prevention of AAD. EVIDENCE ACQUISITION: We did a PubMed, Medline and Cochrane libary search for literature available in last 25 years. RESULTS: Prevalence of antibiotic associated diarrhea (AAD) is around 11% Children younger than 2 years and type of antibiotics are the two risk factors identified for AAD. For the pediatric population, CDI reportedly decreased in a tertiary care hospital in India, though number of suspected samples tested increased. The incidence of community acquired CDI is increasing in the pediatric population also. Detection of toxin A and B by enzyme linked immunosorbent assay (ELISA) and detection of toxin B by tissue culture form the mainstay in the diagnosis of C. difficile. Most of the AAD would respond to only discontinuation or change of the antibiotic. Oral metronidazole or oral vancomycin are drugs of choice for CDI. Probiotics reduce the risk of AAD in children and for every 7-10 patients one less would develop AAD. CONCLUSION: Prevalence of AAD is low and majority will respond to discontinuation of antibiotic. CDI is uncommon in children. Probiotics will prevent AAD in only 1 in 7 children on antibiotics. We need cost effectiveness studies to decide the issue of needing a probiotic antibiotic combination to prevent AAD. PMID- 19556660 TI - Indian slum children in Western cinema: sensitive portrayal of stark reality or crass exploitation? PMID- 19556661 TI - Non-glucose oral rehydration solution does it make a good thing better? PMID- 19556662 TI - Pulmonary alveolar proteinosis. AB - Pulmonary alveolar proteinosis is a rare cause of respiratory distress in neonates. We present a 4 month old infant who presented with progressive respiratory distress since birth and failure to thrive. He was initially treated as a case of diffuse alveolar disease but on open lung biopsy was diagnosed as pulmonary alveolar proteinosis. The child expired at 7 months of age. PMID- 19556663 TI - Chronic eosinophilic leukemia with a unique translocation. AB - We report a case of chronic eosinophilic leukemia in a 9 year old girl who presented with anemia, thrombocytopenia, leucocytosis (mostly dysplastic eosinophils), lymphadenopathy and hepatosplenomegaly. There was no increase in blasts but myelofibrosis was seen in the bone marrow. A previously unreported translocation 46,XX,t(1;4)(q24;q35), was found on cytogenetic analysis and involvement of the myocardium was also present. Shortly after commencing steroids, the family abandoned therapy. PMID- 19556664 TI - Complex regional pain syndrome type 1 and scurvy. AB - A 5 year old female developed features of complex regional pain syndrome (CRPS) i.e excessive pain to touch, decreased sweating and edema of left ankle 2 years after fracture of left tibia. Gum bleeding, petechiae and pseudoparalysis and suggestive radiograph characterized scurvy. Hyperesthesia improved and child walked with support following administration of vitamin C. PMID- 19556665 TI - Carbonic anhydrase II deficiency a novel mutation. AB - Carbonic anhydrase II (CA II) deficiency is an extremely rare autosomal recessive disorder, characterised by a triad of osteopetrosis, renal tubular acidosis and cerebral calcifications. A 12 year old boy with classical features of CA II deficiency is reported who was found to be homozygous for the mutation in CA II gene and parents were heterozygous for the same mutation .To the best of our knowledge this is the first case report of mutation proven CA II deficiency from India. PMID- 19556667 TI - Post varicella thrombosis. PMID- 19556666 TI - Boil to sepsis case of community acquired MRSA. PMID- 19556668 TI - Safe disposal of used sharp objects. PMID- 19556670 TI - Punishment for refusing OPV. PMID- 19556671 TI - Probiotics in clinical practice to use or not to use! PMID- 19556672 TI - Iron folic acid supplementation. PMID- 19556673 TI - Low bone mineral density in childhood acute lymphoblastic leukemia. PMID- 19556676 TI - Hemophagocytic syndrome associated with dengue hemorrhagic fever. PMID- 19556677 TI - Ataxia telangiectasia with hepatocellular carcinoma. PMID- 19556678 TI - Medical calculators: online tool for pediatricians. PMID- 19556679 TI - Decoding human motor activity from EEG single trials for a discrete two dimensional cursor control. AB - This study aims to explore whether human intentions to move or cease to move right and left hands can be decoded from spatiotemporal features in non-invasive EEG in order to control a discrete two-dimensional cursor movement for a potential multidimensional brain-computer interface (BCI). Five naive subjects performed either sustaining or stopping a motor task with time locking to a predefined time window by using motor execution with physical movement or motor imagery. Spatial filtering, temporal filtering, feature selection and classification methods were explored. The performance of the proposed BCI was evaluated by both offline classification and online two-dimensional cursor control. Event-related desynchronization (ERD) and post-movement event-related synchronization (ERS) were observed on the contralateral hemisphere to the hand moved for both motor execution and motor imagery. Feature analysis showed that EEG beta band activity in the contralateral hemisphere over the motor cortex provided the best detection of either sustained or ceased movement of the right or left hand. The offline classification of four motor tasks (sustain or cease to move right or left hand) provided 10-fold cross-validation accuracy as high as 88% for motor execution and 73% for motor imagery. The subjects participating in experiments with physical movement were able to complete the online game with motor execution at an average accuracy of 85.5 +/- 4.65%; the subjects participating in motor imagery study also completed the game successfully. The proposed BCI provides a new practical multidimensional method by noninvasive EEG signal associated with human natural behavior, which does not need long-term training. PMID- 19556681 TI - Fabrication of free-standing replicas of fragile, laminar, chitinous biotemplates. AB - The conformal-evaporated-film-by-rotation technique, followed by the dissolution of chitin in an aqueous solution of orthophosphoric acid, can be used to fabricate free-standing replicas of fragile, laminar, chitinous biotemplates. This novel approach was demonstrated using butterfly wings as biotemplates and GeSeSb chalcogenide glass for replicas. PMID- 19556682 TI - Development of a dual modality imaging system: a combined gamma camera and optical imager. AB - Several groups have reported the development of dual modality Gamma camera/optical imagers, which are useful tools for investigating biological processes in experimental animals. While previously reported dual modality imaging instrumentation usually employed a separated gamma camera and optical imager, we designed a detector using a position sensitive photomultiplier tube (PSPMT) that is capable of imaging both gamma rays and optical photons for combined gamma camera and optical imager. The proposed system consists of a parallel-hole collimator, an array-type crystal and a PSPMT. The top surface of the collimator and array crystals is left open to allow optical photons to reach the PSPMT. Pulse height spectra and planar images were obtained using a Tc-99m source and a green LED to estimate gamma and optical imaging performances. When both gamma rays and optical photon signals were detected, the signal interferences caused by each other signal were evaluated. A mouse phantom and an ICR mouse containing a gamma ray and optical photon source were imaged to assess the imaging capabilities of the system. The sensitivity, energy resolution and spatial resolution of the gamma image acquired using Tc-99m were 1.1 cps/kBq, 26% and 2.1 mm, respectively. The spatial resolution of the optical image acquired with an LED was 3.5 mm. Signal-to-signal interference due to the optical photon signal in the gamma pulse height spectrum was negligible. However, the pulse height spectrum of the optical photon signal was found to be affected by the gamma signal, and was obtained between signals generated by gamma rays with a correction using a veto gate. Gamma ray and optical photon images of the mouse phantom and ICR mouse were successfully obtained using the single detector. The experimental results indicated that both optical photon and gamma ray imaging are feasible using a detector based on the proposed PSPMT. PMID- 19556680 TI - Assessment of gliosis around moveable implants in the brain. AB - Repositioning microelectrodes post-implantation is emerging as a promising approach to achieve long-term reliability in single neuronal recordings. The main goal of this study was to (a) assess glial reaction in response to movement of microelectrodes in the brain post-implantation and (b) determine an optimal window of time post-implantation when movement of microelectrodes within the brain would result in minimal glial reaction. Eleven Sprague-Dawley rats were implanted with two microelectrodes each that could be moved in vivo post implantation. Three cohorts were investigated: (1) microelectrode moved at day 2 (n = 4 animals), (2) microelectrode moved at day 14 (n = 5 animals) and (3) microelectrode moved at day 28 (n = 2 animals). Histological evaluation was performed in cohorts 1-3 at four-week post-movement (30 days, 42 days and 56 days post-implantation, respectively). In addition, five control animals were implanted with microelectrodes that were not moved. Control animals were implanted for (1) 30 days (n = 1), (2) 42 days (n = 2) and (3) 56 days (n = 2) prior to histological evaluation. Quantitative assessment of glial fibrillary acidic protein (GFAP) around the tip of the microelectrodes demonstrated that GFAP levels were similar around microelectrodes moved at day 2 when compared to the 30-day controls. However, GFAP expression levels around microelectrode tips that moved at day 14 and day 28 were significantly less than those around control microelectrodes implanted for 42 and 56 days, respectively. Therefore, we conclude that moving microelectrodes after implantation is a viable strategy that does not result in any additional damage to brain tissue. Further, moving the microelectrode downwards after 14 days of implantation may actually reduce the levels of GFAP expression around the tips of the microelectrodes in the long term. PMID- 19556683 TI - Introducing DeBRa: a detailed breast model for radiological studies. AB - Currently, x-ray mammography is the method of choice in breast cancer screening programmes. As the mammography technology moves from 2D imaging modalities to 3D, conventional computational phantoms do not have sufficient detail to support the studies of these advanced imaging systems. Studies of these 3D imaging systems call for a realistic and sophisticated computational model of the breast. DeBRa (Detailed Breast model for Radiological studies) is the most advanced, detailed, 3D computational model of the breast developed recently for breast imaging studies. A DeBRa phantom can be constructed to model a compressed breast, as in film/screen, digital mammography and digital breast tomosynthesis studies, or a non-compressed breast as in positron emission mammography and breast CT studies. Both the cranial-caudal and mediolateral oblique views can be modelled. The anatomical details inside the phantom include the lactiferous duct system, the Cooper ligaments and the pectoral muscle. The fibroglandular tissues are also modelled realistically. In addition, abnormalities such as microcalcifications, irregular tumours and spiculated tumours are inserted into the phantom. Existing sophisticated breast models require specialized simulation codes. Unlike its predecessors, DeBRa has elemental compositions and densities incorporated into its voxels including those of the explicitly modelled anatomical structures and the noise-like fibroglandular tissues. The voxel dimensions are specified as needed by any study and the microcalcifications are embedded into the voxels so that the microcalcification sizes are not limited by the voxel dimensions. Therefore, DeBRa works with general-purpose Monte Carlo codes. Furthermore, general-purpose Monte Carlo codes allow different types of imaging modalities and detector characteristics to be simulated with ease. DeBRa is a versatile and multipurpose model specifically designed for both x-ray and gamma-ray imaging studies. PMID- 19556684 TI - Collimator optimization in SPECT based on a joint detection and localization task. AB - In SPECT the collimator is a crucial element of the imaging chain and controls the noise-resolution tradeoff of the collected data. Optimizing collimator design has been a long studied topic, with many different criteria used to evaluate the design. One class of criteria is task based, in which the collimator is designed to optimize detection of a signal (lesion). Here we consider a new, more realistic task, the joint detection and localization of a signal. Furthermore, we use an ideal observer-one that attains a theoretically maximum task performance to optimize collimator design. The ideal observer operates on the sinogram data. We consider a family of parallel-hole low-energy collimators of varying resolution and efficiency and optimize over this set. We observe that for a 2D object characterized by noise due to background variability and a sinogram with photon noise, the optimal collimator tends to be of lower resolution and higher efficiency than equivalent commercial collimators. Furthermore, this optimal design is insensitive to the tolerance radius within which the signal must be localized. So for this scenario, the addition of a localization task does not change the optimal collimator. Optimal collimator resolution gets worse as signal size grows, and improves as the level of background variability noise increases. These latter two trends are also observed when the detection task is signal-known exactly and background variable. PMID- 19556685 TI - Systematic analysis of biological and physical limitations of proton beam range verification with offline PET/CT scans. AB - The clinical use of offline positron emission tomography/computed tomography (PET/CT) scans for proton range verification is currently under investigation at the Massachusetts General Hospital (MGH). Validation is achieved by comparing measured activity distributions, acquired in patients after receiving one fraction of proton irradiation, with corresponding Monte Carlo (MC) simulated distributions. Deviations between measured and simulated activity distributions can either reflect errors during the treatment chain from planning to delivery or they can be caused by various inherent challenges of the offline PET/CT verification method. We performed a systematic analysis to assess the impact of the following aspects on the feasibility and accuracy of the offline PET/CT method: (1) biological washout processes, (2) patient motion, (3) Hounsfield unit (HU) based tissue classification for the simulation of the activity distributions and (4) tumor site specific aspects. It was found that the spatial reproducibility of the measured activity distributions is within 1 mm. However, the feasibility of range verification is restricted to a limited amount of positions and tumor sites. Washout effects introduce discrepancies between the measured and simulated ranges of about 4 mm at positions where the proton beam stops in soft tissue. Motion causes spatial deviations of up to 3 cm between measured and simulated activity distributions in abdominopelvic tumor cases. In these later cases, the MC simulated activity distributions were found to be limited to about 35% accuracy in absolute values and about 2 mm in spatial accuracy depending on the correlativity of HU into the physical and biological parameters of the irradiated tissue. Besides, for further specific tumor locations, the beam arrangement, the limited accuracy of rigid co-registration and organ movements can prevent the success of PET/CT range verification. All the addressed factors explain why the proton beam range can only be verified within an accuracy of 1-2 mm in low-perfused bony structures of head and neck patients for which an accurate co-registration of predominant bony anatomy is possible, as shown previously. However, most of the limitations of the current approach are conquerable. By implementing technological and methodological improvements like the use of in-room PET scanners, PET measurements could soon be used to provide proton range verification in clinical routine. PMID- 19556686 TI - An image-based skeletal tissue model for the ICRP reference newborn. AB - Hybrid phantoms represent a third generation of computational models of human anatomy needed for dose assessment in both external and internal radiation exposures. Recently, we presented the first whole-body hybrid phantom of the ICRP reference newborn with a skeleton constructed from both non-uniform rational B spline and polygon-mesh surfaces (Lee et al 2007 Phys. Med. Biol. 52 3309-33). The skeleton in that model included regions of cartilage and fibrous connective tissue, with the remainder given as a homogenous mixture of cortical and trabecular bone, active marrow and miscellaneous skeletal tissues. In the present study, we present a comprehensive skeletal tissue model of the ICRP reference newborn to permit a heterogeneous representation of the skeleton in that hybrid phantom set-both male and female-that explicitly includes a delineation of cortical bone so that marrow shielding effects are correctly modeled for low energy photons incident upon the newborn skeleton. Data sources for the tissue model were threefold. First, skeletal site-dependent volumes of homogeneous bone were obtained from whole-cadaver CT image analyses. Second, selected newborn bone specimens were acquired at autopsy and subjected to micro-CT image analysis to derive model parameters of the marrow cavity and bone trabecular 3D microarchitecture. Third, data given in ICRP Publications 70 and 89 were selected to match reference values on total skeletal tissue mass. Active marrow distributions were found to be in reasonable agreement with those given previously by the ICRP. However, significant differences were seen in total skeletal and site-specific masses of trabecular and cortical bone between the current and ICRP newborn skeletal tissue models. The latter utilizes an age independent ratio of 80%/20% cortical and trabecular bone for the reference newborn. In the current study, a ratio closer to 40%/60% is used based upon newborn CT and micro-CT skeletal image analyses. These changes in mineral bone composition may have significant dosimetric implications when considering localized marrow dosimetry for radionuclides that target mineral bone in the newborn child. PMID- 19556687 TI - A Markov decision process approach to temporal modulation of dose fractions in radiation therapy planning. AB - The current state of the art in cancer treatment by radiation optimizes beam intensity spatially such that tumors receive high dose radiation whereas damage to nearby healthy tissues is minimized. It is common practice to deliver the radiation over several weeks, where the daily dose is a small constant fraction of the total planned. Such a 'fractionation schedule' is based on traditional models of radiobiological response where normal tissue cells possess the ability to repair sublethal damage done by radiation. This capability is significantly less prominent in tumors. Recent advances in quantitative functional imaging and biological markers are providing new opportunities to measure patient response to radiation over the treatment course. This opens the door for designing fractionation schedules that take into account the patient's cumulative response to radiation up to a particular treatment day in determining the fraction on that day. We propose a novel approach that, for the first time, mathematically explores the benefits of such fractionation schemes. This is achieved by building a stylistic Markov decision process (MDP) model, which incorporates some key features of the problem through intuitive choices of state and action spaces, as well as transition probability and reward functions. The structure of optimal policies for this MDP model is explored through several simple numerical examples. PMID- 19556688 TI - Development of a positron probe for localization and excision of brain tumours during surgery. AB - The survival outcome of patients suffering from gliomas is directly linked to the complete surgical resection of the tumour. To help the surgeons to delineate precisely the boundaries of the tumour, we developed an intraoperative positron probe with background noise rejection capability. The probe was designed to be directly coupled to the excision tool such that detection and removal of the radiolabelled tumours could be simultaneous. The device consists of two exchangeable detection heads composed of clear and plastic scintillating fibres. Each head is coupled to an optic fibre bundle that exports the scintillating light to a photodetection and processing electronic module placed outside the operative wound. The background rejection method is based on a real-time subtraction technique. The measured probe sensitivity for (18)F was 1.1 cps kBq( 1) ml(-1) for the small head and 3.4 cps kBq(-1) ml(-1) for the large head. The mean spatial resolution was 1.6 mm FWHM on the detector surface. The gamma-ray rejection efficiency measured by realistic brain phantom modelling of the surgical cavity was 99.4%. This phantom also demonstrated the ability of the probe to detect tumour discs as small as 5 mm in diameter (20 mg) for tumour-to background ratios higher than 3:1 and with an acquisition time around 4 s at each scanning step. These results indicate that our detector could be a useful complement to existing techniques for the accurate excision of brain tumour tissue and more generally to improve the efficiency of radio-guided cancer surgery. PMID- 19556690 TI - Histochemical and immunohistochemical characterization of exocrine cells in the foregut of the red-eared slider turtle, Trachemys scripta (Emydidae). AB - The morphofunctional organization of the exocrine cells in the foregut of the red eared slider turtle, Trachemys scripta, was investigated by histochemistry (PAS, AB pH1.0 and pH 2.5, HID-AB, Bowie), lectin-histochemistry (WGA, SBA, UEA, ConA, PNA, DBA, sialidase-SBA, sialidase-PNA, Paradoxical ConA), and immunohistochemistry (antipepsin, anti-alpha-H+,K+ ATPase) to detect regional differences and verify the existence of an oro-aboral gradient in gastric juice secretion. Observations showed that pharyngeal goblet cells have mucins with terminal residuals of GalNAc and sialic acid. In the oesophagus, sulphomucins in the goblet cells are progressively substituted by sialomucins and no glands are found. Gastric surface cells secrete mostly sialomucins with residuals of GlcNAc and GalNAc. The cardias presents glands with cells secreting mostly sialomucins, differing from the surface ones in having GalNAcalpha1,3GalNAc sequences. The fundus presents complex glands with main and lesser tubules. Cells in the main tubules secrete stable, class-III mucins with sialylated residuals of glucose and/or mannose, GalNAcalpha1, 3GalNAc, and Galbeta1,3GalNAc sequences. In the lesser tubules, the oxynticopeptic cells are found, presenting pepsinogen granules and an affinity to the anti-alpha-H+,K+ ATPase, without any oro-aboral variation. The pyloric glands have a secretion similar to that of the neck cells of the fundic glands, consisting of stable, class-III mucins mostly sialylated with a high heterogeneity of residuals such as glucose and/or mannose, fucose, GlcNAc, and GalNAc. Mucins in the foregut are probably involved in several functions such as lubrication, protection against gastric juice, osmotic regulation to increase intestinal absorption, and protection against microbial injuries. PMID- 19556689 TI - Induced current magnetic resonance electrical impedance tomography of brain tissues based on the J-substitution algorithm: a simulation study. AB - We have investigated induced current magnetic resonance electrical impedance tomography (IC-MREIT) by means of computer simulations. The J-substitution algorithm was implemented to solve the IC-MREIT reconstruction problem. By providing physical insight into the charge accumulating on the interfaces, the convergence characteristics of the reconstruction algorithm were analyzed. The simulation results conducted on different objects were well correlated with the proposed theoretical analysis. The feasibility of IC-MREIT to reconstruct the conductivity distribution of head-brain tissues was also examined in computer simulations using a multi-compartment realistic head model. The present simulation results suggest that IC-MREIT may have the potential to become a useful conductivity imaging technique. PMID- 19556691 TI - Regional differences in 5-HT receptors in cerebral and testicular arterioles of the rat as revealed by Ca2+ imaging of real-time confocal microscopy: variances by artery size and organ specificity. AB - 5-hydroxytriptamine (5-HT) is an important transmitter for vessel constriction. The present study was performed to clarify the effect of 5-HT on smooth muscles in large- and small-sized cerebral and testicular arterioles by confocal microscopy, with special reference to intracellular Ca2+ concentration ([Ca2+]i) dynamics. In cerebral vessels, 5-HT induced a [Ca2+]i increase and the contraction of smooth muscle cells in large- and midsized arterioles (external diameters>50 microm) but not in small-sized arterioles. Conspicuous [Ca2+]i changes by 5-HT were especially observed in the portions close to the cerebral arterial circle, and the 5-HT-induced responses were caused by both Ca2+ influx and mobilization. Experiments using agonists and antagonists also revealed that cerebral arteriole smooth muscles possess 5-HT1a, 1b, 2 (G-protein-coupled type), and 3 (ion channel type) receptors; specifically, 5-HT2 plays a major role in these responses. On the other hand, in testicular vessels, there were few regional differences among responses to 5-HT, and both large- and small-sized arterioles responded to 5-HT. The responses were caused by only Ca2+ mobilization mediated 5-HT1a and 2. These results indicate that arterioles in different tissues may respond to 5-HT in different manners. Regional differences and the size-dependent manner of responses to 5-HT in cerebral blood vessels also indicate that the regulatory mechanism of blood circulation is highly differentiated in each region of the central nervous system. PMID- 19556692 TI - Comparison between a weight compression and a magnet compression for experimental pressure ulcers in the rat. Histological studies and effects of anesthesia. AB - To develop an experimental model and evaluate the effects of the magnitude and duration of pressure, the rat abdominal wall (25x20 mm) was subjected to compression either by a weight or by two magnets. In the weight compression tests, a steel plate was inserted under anesthesia into the rat peritoneal cavity, and the abdominal wall was compressed in situ between the underlying steel plate and a weight placed on the abdominal wall. This method resulted in moderate changes in the subcutaneous connective tissue and muscle at 100 mmHg (13.3 kPa) for 4 h, while some muscle damage was observed at 50 mmHg (6.7 kPa) for 4 h and at 100 mmHg for 2 or 3 h. In the magnet pinching tests, a magnet was inserted into the peritoneal cavity, and another magnet overlaid on the skin. Then the abdominal wall was compressed by the two magnets with or without anesthesia. The compression without anesthesia produced significant edema and injuries of the abdominal wall at 50 mmHg for 4 h and at 100 mmHg for 3 or 4 h, while the injuries incurred at 100 mmHg for 2 h were mild. Susceptibility to pressure was high in the muscle, moderate in the subcutaneous connective tissue, and low in the skin. The compression with anesthesia produced significantly milder injuries than those under anesthesia. These findings indicate that the difference in the extent of injuries between the weight compression and magnet compression models are clearly attributable to the pentobarbital anesthesia induced during the compression. Results therefore show that experimental pressure ulcers should be examined in a waking condition and that magnet compression is a useful model for studying the pathogenesis of pressure ulcers. PMID- 19556693 TI - The relationship between the cusp pattern and plural stem cell compartments in Guinea pig cheek teeth by chasing BrdU-labeling. AB - Continuously growing rodent incisors have a special epithelial structure for maintaining adult stem cells that shows a bulbous epithelial protrusion at the apical end and is referred to as an "apical bud". Guinea pig cheek teeth (premolars and molars), also continuously growing teeth, have a complex crown shape consisting of plural cusps. The present study clarifies the existence of apical buds in guinea pig premolars/molars as it examines the relationship between the crown shape and the orientation of the apical buds by micro-computed tomography (micro-CT) and immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU). One premolar and three molar teeth in each side of the maxillae and mandibles assumed characteristic features: each horizontally-sectioned tooth showing a complex zigzag shape was composed of a core of dentin covered by a layer of enamel on all axial surfaces except the buccal of the uppers and the lingual of the lowers. Furthermore, four bulbous epithelial protrusions- including the stellate reticulum--were recognized in the apical end of each tooth, where slow-cycling long-term label-retaining cells resided 20 days after a peritoneal injection of BrdU. These data indicate that guinea pig premolars/molars have four apical buds where the epithelial adult stem cells reside. In contrast, rodent incisors, which show a single cone appearance, are covered by enamel on the labial side and possess only one apical bud. The results of this study suggest that plural apical buds, being arranged bucco-lingually and mesio-distally, produce the crown mold in a zigzag fashion. PMID- 19556694 TI - Autonomic and cardiovascular effects of acute high altitude exposure after myocardial infarction and in normal volunteers. AB - BACKGROUND: High sympathetic tone creates a significant risk for ventricular arrhythmias and sudden death, which can especially affect patients after a myocardial infarction (MI) when exercising in a hypoxic environment. METHODS AND RESULTS: The 16 patients after MI and 10 normal volunteers (NV) underwent a 1-day trip from low altitude (540 m, Bern) to high altitude (3,564 m Jungfraujoch, Swiss Alps). Autonomic function under resting and mental stress conditions at low and high altitude was assessed. MI patients demonstrated a significantly lower stroke volume (P<0.05) at rest compared with the NV at low as well as high altitude. High altitude exposure was accompanied by higher low-frequency/high frequency values in the MI patients compared with NV (P<0.01). Following mental stress, MI patients failed to show the normal return to resting values at high altitude, suggesting sustained sympathetic and diminished parasympathetic activation during post-stress recovery. CONCLUSIONS: Although all MI patients were either on beta-blockers or AII inhibitors, they showed higher sympathetic activity during exposure to high altitude compared with NV. In addition, the respective parasympathetic tone was reduced, especially during recovery. This puts patients after MI at an increased risk for cardiac arrhythmias and they should be advised of this if going to high altitudes. PMID- 19556695 TI - Smoking state determined by cotinine and arterial stiffness. AB - BACKGROUND: Exposure to active and passive smoking is associated with arterial stiffness and cotinine is an objective biochemical indicator to classify smoking state. The aim of the study was to survey participant smoking condition with cotinine in North China and discuss its destructive effect on arterial stiffness. METHODS AND RESULTS: Nine hundred and six people were recruited in this study and were classified into non-smokers, passive smokers and active smokers according to the cotinine level. Brachial-ankle pulse wave velocity (baPWV) was measured using an automatic device. Active smokers vs non-smokers showed a significantly higher baPWV (P=0.023) after adjusting for covariates. Participants who smoked longer than 10 years had a higher baPWV than non-smokers (P=0.029) although no significant difference existed between those who had a smoking history of less than 10 years and those who had never smoked. In less developed areas, the active and passive smoking rates were higher than that in Beijing (P<0.001) while active smokers vs non-smokers had a higher baPWV (P=0.017). CONCLUSIONS: Active smokers, as confirmed by the cotinine level have worse arterial stiffness, and long-term smoking can cause marked arterial stiffness. Smoking exposure might bring about more serious effects on arterial function in patients from poor areas than in modernized cities. PMID- 19556696 TI - Expression levels of Toll-like receptor genes in coronary atherosclerotic lesions of patients with acute coronary syndrome or stable angina pectoris. AB - BACKGROUND: The differences between acute coronary syndrome (ACS) and stable angina pectoris (SAP) in Toll-like receptor (TLR) expression levels in coronary plaques are not well known. TLR gene expression levels were examined, not only in peripheral blood mononuclear cells (PBMCs), but also in coronary plaques in ACS and SAP patients. METHODS AND RESULTS: TLR gene expression levels were examined in PBMCs using real-time RT-PCR in 27 ACS patients, 45 SAP patients and 28 control subjects. TLR2 and TLR4 expression levels in the PBMCs were significantly higher in the ACS group than in the SAP group. TLR9 expression levels were not significantly different among the 3 groups. TLR gene expression levels were also measured in directional coronary atherectomy (DCA) samples from 9 ACS and 14 SAP patients. The TLR2 expression levels in the DCA samples did not significantly differ between the 2 groups. The TLR4 expression levels were significantly higher in the ACS group than in the SAP group. CONCLUSIONS: The results suggest that TLR4 signaling could be more associated with plaque destabilization than with plaque progression. TLR4 expression control may be a novel target for ACS treatment. PMID- 19556697 TI - Modified bicaval anastomosis for orthotopic heart transplantation. PMID- 19556698 TI - Cardiovascular disease caused by earthquake-induced stress: psychological stress and cardiovascular disease. PMID- 19556699 TI - Eicosapentaenoic acid (EPA) in reducing secondary cardiovascular events in hypercholesterolemic Japanese patients. PMID- 19556700 TI - Differentiation of endothelial progenitor cells: a useful biomarker? PMID- 19556701 TI - How do poly lactide-co-glycolide microspheres for prostaglandin E1 operate in ischemic tissue? Local sustained release of prostaglandin E1 induces neovascularization in murine hindlimb ischemia. PMID- 19556702 TI - Statin therapy in patients with heart failure and preserved left ventricular function. PMID- 19556704 TI - Isolation and characterization of GoRAV, a novel gene encoding a RAV-type protein in Galegae orientalis. AB - RAV is a unique transcription factor in higher plants with AP2 and B3-like two distinct DNA-binding domains, but its roles in plant growth and development remains unknown. We have isolated a novel RAV family gene from Galegae orientalis, called GoRAV, which responds to cold induction. Sequence alignment showed that it shares high identity with other RAV family members in AP2 and B3 domain. Transient expression analysis using onion epidermal cells indicated that GoRAV protein is localized in the nucleus. Semi-quantitative RT-PCR (S-Q RT-PCR) analysis indicated that GoRAV is induced by cold, dehydration, high-salinity and abscisic acid, with the strongest induction in G. orientalis leaves during the early response to abiotic elicitors. GoRAV is more abundant in leaf than in stem, but is not expressed in root. This work adds a new member to the RAV family. PMID- 19556706 TI - DNA polymorphism in the blast disease resistance gene Pita of the wild rice Oryza rufipogon and its related species. AB - Intra-and interspecific DNA variations in the blast resistance gene Pita in wild rice (Oryza rufipogon), cultivated rice (O. sativa), and two other related wild rice species (O. meridionalis and O. officinalis) were analyzed to elucidate the nucleotide polymorphism maintenance mechanisms and evolution of Pita in these species. Nucleotide diversity at silent sites of O. rufipogon Pita was 0.0101, an intermediate value relative to other O. rufipogon nuclear genes. A dimorphic pattern of nucleotide polymorphism was detected in the O. rufipogon Pita region. Inoculation of the blast fungus Magnaporthe oryzae verified that the O. rufipogon Pita gene resides in a dimorphic sequence type. The resistance Pita allele had lower levels of variation than the susceptibility pita allele. A hypothesis of evolutionary relationships indicated that the amino acid mutation in the O. rufipogon Pita protein responsible for the difference between resistance and susceptibility occurred relatively recently. These results suggested that the resistance Pita originated from the susceptibility pita. Nucleotide diversity at replacement sites of the leucine-rich domain (LRD) of both the resistance and susceptibility O. rufipogon pita was low. In tests of neutrality, significantly negative values were detected for the LRD of O. rufipogon susceptibility pita. The low nucleotide diversity at replacement sites of the LRD of the susceptibility pita could be explained by purifying selection. Comparison of Pita between O. rufipogon and O. officinalis revealed an excess of nonconservative amino acid substitutions in the LRD, which could be related to the host-pathogen interaction. PMID- 19556707 TI - The spatial expression patterns of DROOPING LEAF orthologs suggest a conserved function in grasses. AB - The DROOPING LEAF (DL) gene regulates carpel specification in the flower and midrib formation in the leaf in Oryza sativa (rice). Loss-of-function mutations in the dl locus cause homeotic transformation of carpels into stamens and lack of midrib, resulting in the drooping leaf phenotype. DL is a member of the YABBY gene family and is closely related to the CRABS CLAW (CRC) gene in Arabidopsis thaliana. The function of Arabidopsis CRC, however, differs from that of rice DL: it is responsible for nectary development and is partially involved in carpel identity. Thus, genes related to DL/CRC seem to have functionally diversified during angiosperm evolution. To assess the conservation of DL function in related species, here we examined the in situ expression patterns of DL orthologs in three grass species, i.e., maize, wheat and sorghum, which is assigned to subfamilies different from Ehrhartoideae including O. sativa. The results clearly show that the temporal and spatial expression patterns of DL orthologs in the three species are identical to those of rice DL in both flower and leaf development, suggesting that DL-related genes are functionally conserved within the grass family. It is likely that DL may have been recruited to carpel specification and midrib formation within the lineage of the grass family after divergence of their ancestor from that of eudicots. PMID- 19556708 TI - A Bombyx mandarina mutant exhibiting translucent larval skin is controlled by the molybdenum cofactor sulfurase gene. AB - During the maintenance of the wild silkworm, Bombyx mandarina, a mutant phenotype exhibiting translucent skin was identified. Based on the crossing experiments with the domesticated silkworm, Bombyx mori, we found that the mutant was controlled by molybdenum cofactor sulfurase (MoCoS) gene. We designated the mutant ''Ozaki's translucent'' (og(Z)). We found a 2.1-kb deletion containing the transcription initiation site, exons 1 and 2, and the 5' end of exon 3 of the MoCoS gene. The transcript of the MoCoS gene was not detected in the og(Z) homozygote. We concluded that og(Z) is a complete loss-of-function allele generated by a disruption of the MoCoS gene. PMID- 19556709 TI - Comprehensive phylogeny of the family Sparidae (Perciformes: Teleostei) inferred from mitochondrial gene analyses. AB - Sparid fishes consist of approximately 115 species in 33 genera that are broadly distributed in tropical and temperate coastal waters. Although several phylogenetic analyses were conducted based on specific molecular markers, their classification remains unresolved. Here, we present the most comprehensive molecular phylogeny of the family Sparidae to date, based on cytochrome b (cyt-b) genes. We determined 18 sequences of sparids and conducted phylogenetic analyses among 72 individuals representing 66 sparids with 23 outgroup species. Phylogenetic trees were constructed according to partitioned Maximum Likelihood (ML) and Bayesian methods. The phylogenetic analyses were conducted on two different data sets (including all positions; RY-coding). The phylogenetic trees showed monophyly of the family Sparidae with a different taxon, centracanthid Spicara. The subfamilies in the Sparidae in all trees are non-monophyletic and do not agree with current classification of the subfamilies. The genera Acanthopagrus, Cheimerius, Dentex, Diplodus, Pagellus, Pagrus, and Spicara are also non-monophyletic and their classifications should be revised based on the phylogenetic relationships and reinvestigation of morphological characters. The sparids are divided into three major clades, A, B and C, respectively in the ML tree based on all codon positions, whereas clade C was paraphyletic in the other trees. The species in clade C are known to be present in the eastern Pacific to western Atlantic, whereas those in clades A and B are distributed in various oceanic regions. Some sub-clades in clades A and B consist of species that are distributed in defined local regions. We further investigated evolutionary patterns of 87 morphological characters by ancestral character-state reconstruction according to the parsimony criteria. The results suggested high evolutionary plasticity of the characters in sparids, indicating that it causes species-diversity and taxonomic confusion at various taxonomic levels, and that such convergent evolution may occur more frequently also in other coastal fishes. PMID- 19556710 TI - Physical and functional interaction between WRNIP1 and RAD18. AB - WRN interacting protein 1 (WRNIP1) was originally identified as a protein that interacts with the Werner syndrome responsible gene product (WRN). WRNIP1 is a highly conserved protein from E. coli to humans. Genetic studies in budding yeast suggested that the yeast orthlog of WRNIP1, Mgs1, may function in a DNA damage tolerance pathway that is similar to, but distinct from, the template-switch damage avoidance pathway involving Rad6, Rad18, Rad5, Mms2, and Ubc13. Here we report that human WRNIP1 binds in an ATP dependent manner to both forked DNA that mimics stalled replication forks and to template/primer DNA. We found that WRNIP1 interacts physically with RAD18 and interferes with the binding of RAD18 to forked DNA and to template/primer DNA. In contrast, RAD18 enhances the binding of WRNIP1 to these DNAs, suggesting that WRNIP1 targets DNA bound by RAD18. PMID- 19556711 TI - Phytochrome dependent quantitative control of Hd3a transcription is the basis of the night break effect in rice flowering. AB - A short exposure to light during relative night (night break; NB) delays flowering in the short day plant rice. NB acts by downregulating Heading date 3a (Hd3a) expression. Because phytochrome B mutants do not respond to NB and their flowering time is not affected even under NB conditions, phyB is required for the suppression of Hd3a expression. The effect of NB is quantitatively controlled by light quality and by either light intensity or duration. However, the molecular mechanisms that regulate these interactions are poorly understood. Here, we examine the roles of phytochromes in the regulation of Hd3a transcription under NB conditions using monochromatic red, far-red and blue light. Red and blue light downregulated Hd3a expression, but far-red light NB did not. The effect of red light NB on Hd3a is dependent on photon fluence and is restored by subsequent far red light irradiation. Our results suggest that quantitative effect of light on flowering in rice NB is mediated by the regulation of Hd3a transcription by phyB. PMID- 19556705 TI - Identification of salt-induced genes from Salicornia brachiata, an extreme halophyte through expressed sequence tags analysis. AB - Salinity severely affects plant growth and development causing crop loss worldwide. We have isolated a large number of salt-induced genes as well as unknown and hypothetical genes from Salicornia brachiata Roxb. (Amaranthaceae). This is the first description of identification of genes in response to salinity stress in this extreme halophyte plant. Salicornia accumulates salt in its pith and survives even at 2 M NaCl under field conditions. For isolating salt responsive genes, cDNA subtractive hybridization was performed between control and 500 mM NaCl treated plants. Out of the 1200 recombinant clones, 930 sequences were submitted to the NCBI database (GenBank accession: EB484528 to EB485289 and EC906125 to EC906292). 789 ESTs showed matching with different genes in NCBI database. 4.8% ESTs belonged to stress-tolerant gene category and approximately 29% ESTs showed no homology with known functional gene sequences, thus classified as unknown or hypothetical. The detection of a large number of ESTs with unknown putative function in this species makes it an interesting contribution. The 90 unknown and hypothetical genes were selected to study their differential regulation by reverse Northern analysis for identifying their role in salinity tolerance. Interestingly, both up and down regulation at 500 mM NaCl were observed (21 and 10 genes, respectively). Northern analysis of two important salt tolerant genes, ASR1 (Abscisic acid stress ripening gene) and plasma membrane H+ATPase, showed the basal level of transcripts in control condition and an increase with NaCl treatment. ASR1 gene is made full length using 5' RACE and its potential role in imparting salt tolerance is being studied. PMID- 19556712 TI - Evolutionary characterization of a highly repetitive sequence identified from the false killer whale (Pseudorca crassidens). AB - We report here that a novel 1,869 bp repetitive sequence identified from the false killer whale (Pseudorca crassidens) could be a new molecular phylogenetic marker in cetaceans. Results of PCR amplification and southern blot hybridization using 16 species' genomic DNAs from five different families revealed that the repetitive sequence is highly conserved within all Delphinidae species. Notably, specific primers designed for this repetitive sequence effectively amplified the targeted repetitive units, which were critically dependent upon the genetic phylogenies in the members of the Delphinidae cetaceans. Therefore, the novel sequences can be used as a useful phylogenetic marker for understanding the molecular evolutional studies in members of the Delphinidae family of cetaceans. PMID- 19556713 TI - Atorvastatin Increases Endoglin, SMAD2, Phosphorylated SMAD2/3 and eNOS Expression in ApoE/LDLR Double Knockout Mice. AB - AIM: Endoglin is a homodimeric transmembrane glycoprotein that has been demonstrated to affect transforming growth factor beta (TGF-beta) signaling and endothelial nitric oxide synthase (eNOS) expression by affecting SMAD proteins in vitro. Thus, in this study we stepped forward to elucidate whether endoglin is co expressed with SMAD2, phosphorylated SMAD2/3 proteins and eNOS in vivo in atherosclerotic lesions in ApoE/LDLR double knockout mice. In addition, we sought whether endoglin expression as well as the expression of SMAD2, phosphorylated SMAD2/3 and eNOS is affected by atorvastatin treatment. METHODS: Two-month-old female ApoE/LDLR double knockout mice were divided into two groups. The control group was fed with the western type diet whereas in the atorvastatin group, atorvastatin at dose 100 mg/kg per day was added to the same diet. Immunohistochemical and western blot analysis of endoglin, SMAD2, phosphorylated SMAD2/3 and eNOS expressions in aorta were performed. RESULTS: The biochemical analysis showed that administration of atorvastatin significantly decreased level of total cholesterol, VLDL, LDL, TAG, and significantly increased level of HDL cholesterol. Fluorescence immunohistochemistry showed endoglin co-expression with SMAD2, phosphorylated SMAD2/3 and eNOS in aortic endothelium covering atherosclerotic lesions in both control and atorvastatin treated mice. Western blot analysis demonstrated that atorvastatin significantly increased expression of endoglin, SMAD2, phosphorylated SMAD2/3, and eNOS in mice aorta. CONCLUSION: These findings suggest, that endoglin might be interesting marker of endothelial dysfunction and/or atherogenesis which is upregulated by statins implicating potential beneficial role of endoglin and its pathway in atherosclerosis. PMID- 19556714 TI - Influence of statins on glucose tolerance in patients with type 2 diabetes mellitus: subanalysis of the collaborative study on hypercholesterolemia drug intervention and their benefits for atherosclerosis prevention (CHIBA study). PMID- 19556715 TI - Association of body mass index with coronary plaque regression: 6-month prospective study. AB - AIM: Obesity is a well known strong risk factor for coronary artery disease (CAD). We prospectively investigated the influence of body mass index (BMI) on the inhibitory effects of pravastatin against the development of coronary atherosclerosis. METHODS: In 56 patients with stable CAD, 3-dimensional intravascular ultrasound was performed in matched coronary segments at the baseline and after 6-month treatment with pravastatin. RESULTS: The plaque volume was significantly reduced by 11% after treatment (p<0.001 vs. baseline). The percent plaque volume was positively correlated with the baseline BMI (r=0.37, p<0.001), and negatively correlated with the serum total cholesterol / high density lipoprotein cholesterol ratio (r=0.27, p<0.05) and total leukocyte count (r=0.27, p<0.05). Multivariate regression analysis showed that BMI was an independent predictor of the change in plaque volume (beta coefficient: 0.326; 95% CI: 0.003 to 0.037; p<0.05). No correlations were found between BMI and changes in the serum levels of any other lipids, apolipoproteins, or hs-CRP. CONCLUSION: The present study demonstrated that an increase in BMI attenuated pravastatin-induced coronary atherosclerosis regression. The results may provide new insight into the framework for the treatment of obese patients with CAD. PMID- 19556716 TI - Genotypic Effect of ABCG1 gene promoter -257T>G polymorphism on coronary artery disease severity in Japanese men. AB - AIM: ATP-binding cassette transporter G1 (ABCG1) is a cholesterol transporter that plays a role in cholesterol efflux in the presence of high-density lipoprotein (HDL). Moreover, HDL-mediated cholesterol efflux is increased in cultured ABCG1 overexpressed cells; however, the physiological roles of ABCG1 and its contribution to atherosclerosis in humans remain unclear. METHODS: The effect of ABCG1 -257T>G mutation on transcription activity was determined by a reporter assay. One hundred nine Japanese men with coronary artery disease (CAD) were analyzed. ABCG1 -257T>G polymorphism was assessed by mutation-selective PCR methods to identify T/T, T/G, and G/G genotypes. RESULTS: The reporter assay showed that ABCG1 transcription activity was significantly lower (p<0.01) in the G allele of -257T>G polymorphism compared with that in the T allele. Clinically, there were no significant differences in serum triglyceride and total, LDL, and HDL cholesterol levels, or other risk factors. Logistic regression analysis revealed significant additive and dominant effects on the frequency of patients with multi-vessel disease compared with single-vessel disease (T/T vs. T/G vs. G/G, odds ratio:2.1, p=0.027; T/T vs. T/G and. G/G, odds ratio:3.5, p=0.005). CONCLUSION: This is the first report to show that a novel ABCG1 -257T>G promoter polymorphism influences CAD severity in Japanese men. PMID- 19556718 TI - Serum amyloid A protein and carotid intima-media thickness in healthy young subjects. PMID- 19556717 TI - HMW-adiponectin associates with triglyceride concentrations in type 1 diabetic patients. AB - AIM: The objective of this study was to investigate the relation between serum high molecular weight (HMW)-adiponectin concentration and fasting and postprandial blood glycemic and lipid parameters in type 1 diabetic patients. METHODS: Type 1 diabetic patients treated with short-acting insulin analogs and healthy volunteers were recruited. The subjects were divided into two groups based on the mean HMW-adiponectin value of 12.2 mg/L: low HMW-adiponectin (men/women=7/2) and high HMW-adiponectin (men/women=3/8). Blood samples were collected after an overnight fast, and 30180 min after consuming white bread (B) or white bread with butter (BB). RESULTS: Type 1 diabetic patients with high HMW adiponectin had lower triglyceride and remnant like particle (RLP)-triglyceride concentrations than those with low HMW-adiponectin (p<0.01), and had a higher lipoprotein lipase mass (p<0.01) than healthy subjects (men/women=8/6). After B and BB meals, type 1 diabetic patients with high HMW-adiponectin consistently had lower triglyceride and RLP-triglyceride concentrations for up to 180 min than those with low HMW-adiponectin (p<0.01); however, apoB48 did not differ between these two groups. Fasting and postprandial plasma glucose were higher in both diabetic groups than in healthy subjects (p<0.001), with no significant difference between patient groups. CONCLUSION: HMW-adiponectin is more strongly associated with very low density lipoprotein remnant metabolism than glucose utilization in type 1 diabetic patients receiving short-acting insulin analogs. PMID- 19556719 TI - Circadian variations of gastrointestinal mucosal damage detected with transnasal endoscopy in apparently healthy subjects treated with low-dose aspirin (ASA) for a short period. AB - AIM: In this study, transnasal endoscopy was used to observe the time-course changes of gastrointestinal mucosa with low-dose aspirin (ASA), and the preventive effect of rebamipide was compared with placebo. METHODS: Twenty healthy H.pylori-negative subjects were divided between those receiving 100 mg aspirin with placebo, and those receiving 100 mg aspirin+300 mg rebamipide for 7 days daily. Transnasal endoscopy was performed at 0, 2, 6 and 24 hrs on the first day, and then on the third and seventh days. RESULTS: Ulcers, in the duodenum at 24 hrs and in the antrum at 72 hrs, improved with continuous ASA. Erosions were mainly observed in the duodenum; erosions amounted to 14 at 3 days and 19 at 7 days in the placebo group. No ulcers and some erosions were mainly observed in the duodenum; erosions amounted to 5 at 3 days and 3 at 7 days in the rebamipide group. CONCLUSION: In short-term gastrointestinal damage induced by ASA, damage was observed in the duodenum most frequently, and peak damage was at 24 hrs and 72 hrs. Almost all damage improved gradually in spite of continuous ASA. Rebamipide reduced the damage of low-dose aspirin-induced GI complications. PMID- 19556720 TI - Involvement of vascular angiotensin II-forming enzymes in the progression of aortic abdominal aneurysms in angiotensin II- infused ApoE-deficient mice. AB - AIM: Angiotensin (Ang) II-induced abdominal aortic aneurysm (AAA) in apoE deficient mice has been used as a model of human AAA, but it has been unclear why the progression of AAA continues after stopping the Ang II infusion. The involvement of vascular Ang II-forming enzymes in the progression of AAA was studied. METHODS: ApoE-deficient mice were infused with Ang II (1,000 ng/kg/min) for 4 weeks and evaluated until 20 weeks after the Ang II infusion. Just after and 20 weeks after stopping the Ang II infusion, the degree of AAA, as well as the ACE and chymase activities, was evaluated. An Ang II receptor blocker (candesartan, 30 mg/kg/day) and an angiotensin-converting enzyme (ACE) inhibitor (lisinopril, 60 mg/kg/day) were given for 20 weeks after stopping the Ang II infusion. RESULTS: The aortic diameter expanded just after stopping the Ang II infusion and progressed for a further 20 weeks after the infusion was stopped. Just after stopping the infusion, aortic ACE and chymase activities were significantly increased, but only the increase in chymase activity continued until 20 weeks after the infusion was stopped. Candesartan and lisinopril significantly attenuated aortic diameter expansion. CONCLUSION: The increases in vascular Ang II-forming activities were involved in the progression of AAA after stopping the Ang II infusion. PMID- 19556722 TI - Remnant lipoproteins as strong key particles to atherogenesis. AB - Recent epidemiologic studies have revealed that hypertriglyceridemia is associated with atherosclerosis independent of other coronary risk factors. However, it is difficult to select patients at high risk for coronary artery disease using only serum triglyceride levels compared with low-density lipoprotein cholesterol levels since multiple factors are associated with elevating triglycerides. Atherosclerotic diseases with high triglyceride levels can be found in patients with familial combined hyperlipidemia, diabetes mellitus, and metabolic syndrome, in which remnant lipoproteins accumulate in the circulating blood. Recent researches have paid attention to remnant lipoproteins as atherogenic particles with the development of methods for measuring remnant cholesterol levels and apolipoprotein B-48 levels directly from human serum. Measurement of these parameters in addition to serum triglycerides may help to distinguish high-risk patients and enable us to prevent or suppress the progression of atherosclerotic diseases in those patients. However, questions remain to be answered to evaluate the significance of remnant lipoproteins. Here, we focus on three issues: the underlying problems in measuring remnant lipoprotein cholesterol, the assessment of postprandial hyperlipidemia as an atherogenic condition, and finally a review of our experimental and clinical findings about the mechanisms by which remnant lipoproteins induce atherosclerosis. PMID- 19556721 TI - Impaired insulin secretion in four Tangier disease patients with ABCA1 mutations. AB - AIM: Tangier disease (TD), caused by deficiency of ATP-binding cassette transporter A1, is characterized by the absence of high density lipoprotein and the accumulation of cholesteryl esters in many tissues. Recently, it has been reported that ABCA1 is expressed in pancreatic beta cells and mice with specific inactivation of ABCA1 in beta cells showed markedly impaired insulin secretion, suggesting that ABCA1 deficiency may be involved in diabetes. The aim of the current study was to confirm these findings by the oral glucose tolerance test (OGTT) in human subjects with ABCA1 deficiency. METHODS AND RESULTS: Four Japanese patients with TD were investigated by OGTT with 75 g glucose. In all TD patients, the plasma glucose concentration after 30 min progressively increased, indicating a type 2 diabetic pattern; however the plasma insulin concentration did not respond well to glucose increase. The calculated insulinogenic index was significantly lower in TD patients than in non-diabetic controls (0.055+/-0.034 vs 0.775+/-0.538, mean+/-SD, p<0.05, respectively). CONCLUSIONS: Although the number of TD patients was very small in the current study, these observations indicated a possible mechanism that glucose-stimulated insulin secretion might be impaired in human TD patients with ABCA1 mutations. Taken together, ABCA1 may be involved in insulin secretion from pancreatic beta-cells. PMID- 19556723 TI - S447X polymorphism in the lipoprotein lipase gene and the adiponectin level in the general population: results from the Mima study. AB - AIM: Lipoprotein lipase (LPL) plays an important role in lipid metabolism. There is an association between the common S447X polymorphism in the LPL gene and high density lipoprotein cholesterol (HDL-C) levels, and some association between circulating HDL-C and adiponectin levels has been suggested; however, it is not known whether there is any association between the S447X polymorphism and adiponectin levels. The aim of this study was to investigate whether the LPL S447X polymorphism was associated with adiponectin in the general population. METHODS: LPL S447X was analyzed in 277 community-dwelling subjects (123 men and 154 women, mean age; 65+/-13 years) in the Mima study. Whole samples were genotyped using a fluorescent allele-specific DNA primer assay system. The allele frequencies and any associations with serum lipid and adiponectin levels were investigated. RESULTS: The allele frequencies were S=0.875 and X=0.125 for the LPL S447X polymorphism. The carriers of the X allele had significantly higher levels of adiponectin and HDL-C than non-carriers. The presence of the X allele was significantly associated with higher adiponectin levels, independent of age, sex, body mass index, smoking and HDL-C in multiple regression analyses. CONCLUSION: The LPL S447X polymorphism might therefore be significantly associated with higher adiponectin levels, independent of HDL-C. PMID- 19556724 TI - Validity and usefulness of aortic arch calcification in chest X-ray. AB - BACKGROUND: Arterial calcification is associated with cardiovascular (CV) disease, to be leading to vessel wall stiffness and causing the management of hemodynamics in the elderly more difficult. Here, we compared the extent of calcification in the aortic arch by reviewing chest X-rays to that in the abdominal aorta as assessed by more detailed examinations. In addition, the validity of the grading and the relationship of this useful grading to clinical risk factors were evaluated. METHODS AND RESULTS: The extent of aortic arch calcification (AAC) on a postero-anterior plain chest X-ray was divided into four grades (0 to 3). First, AAC grade was assessed in patients who underwent two quantitative examinations for abdominal aortic calcification; lateral radiograph of lumbar spine and/or computer tomography, and was positively correlated with the abdominal aortic calcification level. Subsequently, AAC grade in 239 out patients (115 men; mean age, 61.9 years) was also evaluated, and was 0, 1, 2, and 3 in 46%, 22%, 29%, and 4% of the population, respectively, was significantly associated with pulse pressure and intima-media thickness. AAC grade in patients with diabetes or renal dysfunction was significantly higher than in those without each risk, but there was no association with other risk factors. In addition, AAC grade was positively correlated with risk factor clustering. CONCLUSION: Assessment of AAC detectable on a chest X-ray is very useful and its grade reflects the magnitude of calcified change in the whole aorta. In addition, AAC evaluation may provide supportive information for atherosclerotic risk stratification. PMID- 19556725 TI - Low incidence of cardiac events in statin-administered patients in CAG study. AB - AIM: The effect of statins in preventing cardiac events in Japanese coronary artery disease (CAD) patients was studied in a retrospective investigation of 148 patients diagnosed with CAD by coronary angiography (CAG). METHODS: Sixty-five patients received statins within 2 weeks after CAG, and 83 patients did not receive statins after CAG. RESULTS: In the statin group, total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) were reduced significantly with statin administration (p<0.01). In the non-statin group, baseline levels of TC, LDL-C and high-density lipoprotein cholesterol were not changed significantly at the end of the follow-up period. As for the effect of statin in preventing cardiac events, the incidence of cardiac events was significantly lower (p<0.0003) in the statin group (n=5: 8%) than in the non-statin group (n=28: 34%). In subanalysis of 37 patients whose TC at the time of initial CAG was less than 200 mg/dL, none of the statin group (n=17) suffered a cardiac event. This was significantly lower than the incidence of cardiac events in the non-statin group (n=5: 25%; p<0.05). CONCLUSION: The present study demonstrated that lowering LDL-C of Japanese CAD patients by statin administration is effective to prevent cardiac events, particularly, a second percutaneous coronary intervention (PCI) for restenosis of a coronary artery following the initial PCI whether or not these patients had hypercholesterolemia. PMID- 19556726 TI - Association of polymorphism of estrogen receptor-alpha gene with circulating levels of adiponectin in postmenopausal women with type 2 diabetes. AB - AIM: Menopause is a risk factor for cardiovascular disease (CVD) in women because of the reduction in endogenous estrogen. Recently, single nucleotide polymorphisms (SNPs) of the estrogen receptor alpha (ESR-1) gene (c.454-397T>C) associated with the prognosis of myocardial infarction in postmenopausal women were identified; however, the mechanism by which genetic variation of ESR-1 contributes to the pathogenesis of CVD is unknown. Circulating levels of adipokines and inflammatory cytokines predict CVD risk; hence, this study aimed to investigate whether ESR-1 genotypes (c.454-397T>C) might influence circulating levels of adipokines and inflammatory cytokines in postmenopausal women with type 2 diabetes. METHODS: Sixty-three postmenopausal women with type 2 diabetes were recruited. Serum levels of adiponectin, resistin, interleukin-6 (IL-6), and high sensitive C-reactive protein (hs-CRP) were determined. RESULTS: The genotype of ESR-1 was closely associated with serum adiponectin, which was decreased in subjects with the T allele and was lowest in those with the T/T genotype. Multiple logistic regression analysis revealed independent contribution of the homozygote for the T allele to low serum levels of adiponectin. CONCLUSION: The T allele of the c.454-397T>C SNP of ESR-1 is associated with low serum levels of adiponectin, which may lead to a high risk of CVD in postmenopausal women. PMID- 19556727 TI - Interaction of fenofibrate and fish oil in relation to lipid metabolism in mice. AB - AIM: The aim of our study is to elucidate the interactive effects on lipid metabolism of fenofibrate and two fish oils with EPA and DHA contents in mice. METHODS: Female C57BL/6J mice were fed purified experimental diets containing safflower oil (SO), EPA-rich menhaden oil (MO) or DHA-rich tuna oil (TO) with or without 0.1% fenofibrate for 8 weeks. At the end of the experiments, we measured plasma lipids and hepatic triglycerides and cholesterol, and the hepatic mRNA expression of lipogenic and lipidolytic genes. RESULTS: Plasma TG levels fell in the group fed MO or TO alone and fell significantly in all fenofibrate-treated groups. Although plasma total cholesterol levels fell significantly in fish oil fed groups, fenofibrate treatments increased significantly plasma total cholesterol levels in these fish oil groups, but not in the group fed SO alone; however, hepatic triglyceride and total cholesterol levels markedly decreased in MO-or TO-fed mice. In lipid synthesis, the hepatic mRNA level of SREBP-1c was not reduced in either fish oil group; however, Insig-1 mRNA decreased in MO and TO feeding groups by about half and FAS or SCD-1 mRNA decreased significantly in MO and TO feeding groups, compared with the SO feeding group. In both fish oil groups, SREBP-2 mRNA decreased significantly and HMG-CoA reductase mRNA also decreased with/without fenofibrate. On the other hand, fenofibrate supplementation significantly induced the mRNA expression of AOX and UCP-2, which play a role in lipid catabolism, in all diets. CYP7A1 mRNA increased markedly in mice fed MO diet with fenofibrate, compared with TO diet with fenofibrate. CONCLUSION: These data suggest that differences in dietary contents of EPA and DHA do not influence the inhibition of lipogenesis, and that fenofibrate supplementation stimulates fatty acid oxidation, regardless of the oil type; however, cholesterol catabolism was induced by a combination of EPA-rich fish oil and fenofibrate, which suggests that EPA has a greater synergistic ability for cholesterol catabolism induction by fenofibrate than DHA. PMID- 19556728 TI - Nifedipine interferes with migration of vascular smooth muscle cells via inhibition of Pyk2-Src axis. AB - AIM: Calcium channel blockers (CCBs) inhibit the migration of vascular smooth muscle cells (VSMC) by mechanisms that remain poorly understood. The purpose of the present study was to characterize the signaling mechanisms by which CCBs inhibit VSMC migration. METHODS AND RESULTS: Nifedipine potently inhibited platelet-derived growth factor (PDGF)-induced chemotaxis, collagen I-induced haptotaxis, and wound-induced migration of human aortic VSMC. In addition, nifedipine inhibited PDGF-induced membrane ruffling and lamellipodium formation. PDGF-induced VSMC migration was significantly inhibited by PP2, a selective inhibitor of the Src kinase family, and was also significantly inhibited by the expression of kinase-inactive Src, suggesting that Src is required for VSMC migration. Nifedipine also inhibited PDGF-induced Src activation (by 60+/-4% with 30 microM) and tyrosinephosphorylation of Cas, paxillin, and cortactin, which are actin-associated substrates of Src. RNA interference-induced knockdown of the Ca(2+)-dependent tyrosine kinase, Pyk2, resulted in inhibition of PDGF-induced Src activation and migration. Finally, nifedipine inhibited PDGF-induced Pyk2 activation in a dose-dependent manner. CONCLUSION: These data suggest that nifedipine interferes with VSMC migration via inhibition of the Pyk2-Src axis and inhibition of actin remodeling processes, including membrane ruffling and lamellipodium formation. PMID- 19556729 TI - Association of SLC6A9 gene variants with human essential hypertension. AB - AIM: We previously identified a quantitative trait locus (QTL) on rat chromosome 5 that appeared to be primarily controlled by the sympathetic nervous system. Because sympathetic overactivity is related to hypertension, solute carrier family 6, member 9 (SLC6A9) is a candidate gene for the connection of this QTL with blood pressure regulation. In the present study, we therefore explored the role of SLC6A9 genetic variations in human essential hypertension (EH). METHODS: We evaluated three single nucleotide polymorphisms (SNPs) (rs2286245, rs3791124 and rs2486001) in 758 essential hypertension patients and 726 controls. Polymorphism-related genotypes were determined with TaqMan assays. RESULTS: The allelic frequency of rs2286245 (C versus T, p=0.032) showed significant differences between EH and normotensive controls (NT) groups. The genotypic distribution of rs3791124 in its dominant model (AA+GA versus GG, p=0.027) also showed significant differences between EH and NT groups. The genotype and allele distributions of rs2486001 did not exhibit any significant differences. CONCLUSION: We found an association between SLC6A9 gene polymorphisms and essential hypertension in a Japanese population, suggesting that SLC6A9 is a susceptibility locus for essential hypertension. PMID- 19556730 TI - Remote cerebellar hemorrhage (zebra sign) in vascular neurosurgery: pathophysiological insights. AB - Hemorrhage in regions remote from the site of initial intracranial operations is rare, but may be fatal. Postoperative cerebellar hemorrhage as a complication of supratentorial surgery, with a radiological appearance known as zebra sign, is an increasingly recognized clinical entity and is associated mainly with vascular neurosurgery or temporal lobe resection. The pathophysiology remains unclear. Three cases of remote cerebellar hematoma occurred after neck clipping of anterior communicating artery aneurysms. All patients had similar clinical findings and underwent pterional craniotomy with the head in accentuated extension. One patient died and the two were discharged without symptoms. Cerebellar hemorrhage probably has a multifactorial origin involving positioning associated with abundant cerebrospinal fluid drainage causing cerebellar sag with resultant vein stretching and bleeding, and use of aspirin or other antiplatelet agents. PMID- 19556731 TI - Cilostazol may prevent cerebral vasospasm following subarachnoid hemorrhage. AB - Cilostazol is an antiplatelet aggregation inhibitor drug associated with increased cerebral blood flow and inflammation suppression. This study evaluated administration of cilostazol to prevent cerebral vasospasm following subarachnoid hemorrhage (SAH) in 50 patients treated surgically from December 2004 to November 2006. All patients, excluding those with Hunt and Kosnik grade 5 or who had undergone late surgery, were classified into two groups: 26 patients who received 200 mg/day cilostazol from postoperative day 1 to day 14 and 24 control patients. The frequency and the degree of cerebral vasospasm, occurrence of ischemic lesion, and clinical symptoms due to vasospasm were compared between the two groups. The appearance of severe vasospasm on angiography, persistent symptomatic spasm, and new cerebral infarction due to vasospasm demonstrated by neuroimaging were apparently lower in the cilostazol group than in the control group, suggesting that cilostazol may significantly suppress cerebral vasospasm following SAH. PMID- 19556732 TI - Effect of intervertebral disk degeneration on spinal stenosis during magnetic resonance imaging with axial loading. AB - Magnetic resonance (MR) imaging with axial loading can simulate the physiological standing state and disclose spinal stenosis undetected or underestimated in the conventional position. Intervertebral disk degeneration may be an important factor in spinal stenosis. This study investigated whether intervertebral disk degeneration increases spinal stenosis during axial loading. MR imaging with and without axial loading was obtained in 51 patients with neurogenic intermittent claudication and/or sciatica and reviewed retrospectively. The grade of disk degeneration was rated in four disk spaces from L2-3 to L5-S1. The dural sac cross-sectional area (DCSA) was measured on MR images taken in both conventional and axial loading positions, and the change in the DCSA was calculated. The effect of disk degeneration on the DCSA was statistically analyzed. Significant decreases in the DCSA occurred with grade 4 disk degeneration (mean +/- standard deviation, 20.1 +/- 14.1 mm(2)), followed by grade 3 (18.3 +/- 15.1 mm(2)) and grade 2 (8.9 +/- 13.1 mm(2)). DCSA decreased considerably with increased severity of disk degeneration with axial loading, except for grade 5 disk degeneration. More accurate diagnosis of stenosis can be achieved using MR imaging with axial loading, especially if grade 2 to 4 disk degeneration is present. PMID- 19556734 TI - Tension pneumocephalus as a complication of lumbar drainage for cerebral aneurysm surgery--case report. AB - A 66-year-old man developed tension pneumocephalus after failed lumbar drainage before clipping surgery for a ruptured anterior communicating artery aneurysm. After puncture with a Tuohy needle, the spinal catheter could not be inserted into the spinal dura, so surgery proceeded without the catheter placement. The patient's neurological status deteriorated suddenly into coma within 15 hours after uneventful clipping of the aneurysm. Computed tomography revealed tension pneumocephalus with marked brain shift. Intracranial hypotension was probably caused by continuous cerebrospinal fluid leakage from the iatrogenic spinal dural tear, resulting in air entry and accumulation into the cranium from an unidentified opening of the cranial dura. The patient was immediately treated with autologous epidural blood patch administration in the lumbar spine, followed by reopening of the craniotomy incision and flap to evacuate the accumulated air. The patient made a quick and uneventful neurological recovery after the rescue procedure. PMID- 19556733 TI - Susceptibility-weighted magnetic resonance imaging detects impaired cerebral hemodynamics in the superior sagittal sinus thrombosis--case report. AB - An 11-year-old female receiving treatment for acute lymphoblastic leukemia presented with superior sagittal sinus (SSS) thrombosis. T(1)-weighted, T(2) weighted, and fluid-attenuated inversion recovery magnetic resonance (MR) imaging, and MR venography showed that the SSS was totally occluded by thrombus. Susceptibility-weighted MR imaging showed hypointense thrombus in the SSS and markedly dilated cortical veins over the bilateral cerebral hemispheres. Two days later, her symptoms had slightly resolved. Iodine-123 N-isopropyl-p iodoamphetamine single photon emission computed tomography showed marked decrease of cerebral blood flow in the bilateral frontal lobes, indicating that venous congestion had disturbed the cerebral hemodynamics. MR venography showed that the SSS was still mostly occluded, but susceptibility-weighted imaging showed that the dilation of the cortical veins was less marked, suggesting that collateral venous routes had gradually developed. The finding of dilated cortical veins had almost disappeared at 28 days after the onset. Susceptibility-weighted imaging can be used as a non-invasive method to monitor the severity of venous congestion caused by cerebral venous sinus thrombosis. PMID- 19556735 TI - Intratumoral hemorrhage of spinal schwannoma of the cauda equina manifesting as acute paraparesis--case report. AB - A 64-year-old man presented with spinal schwannoma of the cauda equina at the thoraco-lumbar junction manifesting as acute paraparesis and urinary incontinence after intratumoral hemorrhage. Surgical resection of the tumor resulted in significant neurological recovery. T(2)(*)-weighted gradient-echo magnetic resonance imaging is very useful to help establish the preoperative diagnosis, leading to early surgical intervention and better outcome after surgery. PMID- 19556736 TI - Malignant intracranial meningioma with spinal metastasis--case report. AB - A 48-year-old woman presented with a left cerebellopontine angle mass. Over a 93 month period, the patient underwent seven surgeries, two radiosurgeries, and one external beam radiotherapy. The tumor was histologically benign at the first operation, but exhibited unusually aggressive behavior after failed radiosurgery and demonstrated clinical characteristics of malignancy such as spinal metastasis. The patient underwent gamma knife radiosurgery (GKR) for recurrence after the first operation, despite the tumor being located in a resectable region. The tumor did not respond. Six months after the sixth surgery, the patient presented with progressive lower extremity paraparesis and sensory disturbance below the T11 dermatome. Magnetic resonance imaging revealed multiple intradural mass lesions located at the T2, T11-T12, and L2 levels. She died 4 months after the diagnosis of spinal metastases. Retrospectively, we speculate that if a tumor is located in a resectable region and Simpson grade I or II tumor resection is possible, direct surgery may be a safer option than GKR. PMID- 19556737 TI - Intramedullary spinal cord abscess treated with antibiotic therapy--case report and review. AB - A 58-year-old man presented with an intramedullary spinal cord abscess (ISCA) manifesting as posterior neck pain, gait disturbance, and urinary retention, and transverse myelopathy 1 week later. Magnetic resonance imaging showed the ISCA at the C7 to T1 levels. He was treated under a diagnosis of cryptogenic ISCA with high-dose ampicillin and third- or fourth-generation cephalosporins, which resulted in complete recovery after 2 months. Review of the literature between January 1998 and August 2007 identified 26 cases of ISCA, including our patient. We also identified two additional nonsurgically treated ISCA patients reported between 1977 and 2007. The most common presentation was motor deficits in all patients, followed by fever, pain, and bladder dysfunction. The mortality rate was 1 of 26 patients, and neurological sequelae were observed in 15 of the 25 surviving patients. There was no significant difference in the frequency of neurological sequelae between surgically and nonsurgically treated patients. Mean length of the abscess in the surgically treated group was significantly larger than that in the medically treated group (5.8 vs. 2.2 vertebral bodies). All three nonsurgically treated patients with neurological sequelae had anaerobic infections and received antibiotic therapy later and for shorter periods than those with complete neurological recovery. Antibiotic treatment is comparable to surgery plus antibiotic treatment. Early broad-spectrum high-dose ampicillin and third-generation cephalosporin, covering Gram-positive, Gram-negative, and anaerobic organisms, should be the first choice of management for patients with ISCA. PMID- 19556738 TI - Transethmoidal meningoencephalocele involving the olfactory bulb with enlarged foramina of the lamina cribrosa--case report. AB - A 3-year-old girl presented with a transethmoidal meningoencephalocele manifesting as recurrent rhinorrhea. Initially, she developed meningitis, but after treatment she experienced rhinorrhea. Two months later, she again presented with rhinorrhea. Neuroimaging studies revealed a small protrusion (15 mm x 10 mm) at the roof of the ethmoidal sinus. Nasal endoscopy confirmed the diagnosis of meningoencephalocele. The operative findings revealed a small hole in the left olfactory bulb, which had descended into an enlarged foramen along with the arachnoid membrane. The left olfactory bulb was removed, and the enlarged foramina of the lamina cribrosa were covered with a frontal pericranial flap. The defect in the bone was very small, but contributed to the development of meningitis and leakage of the cerebrospinal fluid. Basal cephalocele should be considered in a patient with recurrent rhinorrhea and intracranial infections, even in the absence of any apparent anomaly. PMID- 19556739 TI - Concept of ideal closure line for clipping of middle cerebral artery aneurysms- technical note. AB - The concept of optimum closure line was applied to a series of 51 consecutive middle cerebral artery aneurysms (14 ruptured, 37 unruptured) in 41 patients, 16 men and 25 women aged 29-79 years (mean 59.1 years). Visual inspection through the operating microscope revealed 3 types of aneurysm based on the origin of the aneurysm: bifurcation type (n = 39), trunk type (n = 9), and combined type (n = 3). Clipping along the optimum closure line should restore the vascular structure to the original configuration. Combination clip techniques were useful to form a curved closure line. This technique requires adequate operative fields with dissection of the aneurysm and related arteries from the neighboring structures as far as possible. The closure line concept is helpful to decide how to apply clips for particular aneurysms to avoid risks of ischemic complication and future recurrence. Combination clip techniques are often necessary to match a curved closure line. PMID- 19556740 TI - Genetic polymorphisms of the serotonin transporter, but not the 2a receptor or nitric oxide synthetase, are associated with pulmonary hypertension in chronic obstructive pulmonary disease. AB - BACKGROUND: Pulmonary hypertension (PH) is prognosti- cally important in chronic obstructive pulmonary disease (COPD). Since PH only weakly correlates with hypoxemia, other factors must play a role. OBJECTIVE: To investigate whether polymorphisms of the serotonin transporter (5HTT), serotonin-2a receptor (5HTR2a) and endothelial nitric oxide synthetase (eNOS) are related to PH in COPD. METHODS: In 59 COPD patients who underwent right heart catheterization, 6-min walking distance, NYHA functional class, pulmonary function tests, blood gases and 5HTT, 5HTR2a and eNOS (4ab and T298C) polymorphisms were determined. RESULTS: Forty-nine COPD patients in NYHA functional class III-IV were included. Ten were excluded due to comorbid causes of PH (mainly chronic thromboembolic). PH (mPAP > or =25 mm Hg) was present in 55% and usually mild, but out of proportion (mPAP > or =40 mm Hg) in 12%. Patients with PH had significantly higher frequencies of the 5HTT-L-allele (52%) compared to individuals without PH (36%), and LL homozygote patients had more severe PH. In patients with out-of-proportion PH, the L-allelic frequency was even 75%. We found no association of 5HTR2a and eNOS polymorphism with PH in COPD. CONCLUSIONS: In this COPD cohort we confirm that PH is frequent and usually mild, but out of proportion in a subgroup. We found a significant association of the L-allelic variant of 5HTT with PH overall and especially in out-of-proportion PH. These findings may point towards a role of the serotonin system in COPD-PH and warrant further studies. PMID- 19556741 TI - Increased bronchoalveolar lavage fluid CD1c expressing dendritic cells in idiopathic pulmonary fibrosis. AB - BACKGROUND: Chronic inflammation is implicated in idiopathic pulmonary fibrosis (IPF) pathogenesis and is associated with persistent activation of immune responses. These are largely controlled by dendritic cells (DCs). Although large numbers of DCs infiltrate the lungs of patients with IPF, there are no similar reports in bronchoalveolar lavage fluid (BALF). OBJECTIVES: We aimed to investigate DC populations in BALF of IPF patients. METHODS: CD1c(+) myeloid DCs, BDCA3(high) myeloid DCs, BDCA2(+) plasmacytoid DCs and CD83(+) mature DCs were identified by flow cytometry in the BALF of 10 IPF patients and 10 controls. DC numbers were expressed as percentages of total BALF leukocytes. RESULTS: CD1c(+) myeloid DCs were increased in IPF patients versus controls [median (ranges in parentheses) 1.16% (0.25-3.97) vs. 0.61% (0.19-1.10), p = 0.01]. There was also a trend towards increased BDCA3(high) myeloid DCs [0.57% (0.23-0.88) vs. 0.28% (0.07-0.96), p = 0.07]. No differences were reported in BDCA2(+) DCs and CD83(+) DCs between IPF patients and controls. CONCLUSIONS: IPF is associated with an increase in percentages of BALF myeloid DCs. Considering that such an increase was not observed in CD83(+) mature DCs, most of these DCs should be immature. PMID- 19556742 TI - Histotopographic study of the fibroadipose connective cheek system. AB - The purpose of this study was to investigate the morphology of the superficial musculoaponeurotic system (SMAS). Eight embalmed cadavers were analyzed: one side of the face was macroscopically dissected; on the other side, full-thickness samples of the parotid, zygomatic, nasolabial fold and buccal regions were taken. In all specimens, a laminar connective tissue layer (SMAS) bounding two different fibroadipose connective layers was identified. The superficial fibroadipose layer presented vertically oriented fibrous septa, connecting the dermis with the superficial aspect of the SMAS. In the deep fibroadipose connective layer, the fibrous septa were obliquely oriented, connecting the deep aspect of the SMAS to the parotid-masseteric fascia. This basic arrangement shows progressive thinning of the SMAS from the preauricular district to the nasolabial fold (p < 0.05). In the parotid region, the mean thicknesses of the superficial and deep fibroadipose connective tissues were 1.63 and 0.8 mm, respectively, whereas in the region of the nasolabial fold the superficial layer is not recognizable and the mean thickness of the deep fibroadipose connective layer was 2.9 mm. The connective subcutaneous tissue of the face forms a three-dimensional network connecting the SMAS to the dermis and deep muscles. These connective laminae connect adipose lobules of various sizes within the superficial and deep fibroadipose tissues, creating a three-dimensional network which modulates transmission of muscle contractions to the skin. Changes in the quantitative and qualitative characteristics of the fibroadipose connective system, reducing its viscoelastic properties, may contribute to ptosis of facial soft tissues during aging. PMID- 19556743 TI - Persistent pleuropulmonary air leak treated with autologous blood: results from a university hospital and review of literature. AB - BACKGROUND: Persistent air leak after pulmonary resection is a difficult complication for thoracic surgeons to manage. OBJECTIVES: To show the results of our experience treating persistent pleuropulmonary air leak with autologous blood and review the literature on this specific method of treatment. METHODS: Retrospective study of patients with persistent aerial pleuropulmonary fistula treated with autologous blood. The patient's own blood was collected from a peripheral vein and directly introduced through the pleural drain. An inverted siphon was located in the drainage system to avoid prolonged clamping of the drain. This siphon impeded blood return but not air escape. RESULTS: Between January 2001 and August 2008, 27 patients were treated by the above method. Patient age ranged from 2 to 74 years, and 78% were male. Each procedure used a mean quantity of 92 ml blood. Mean persistent air leak time before pleurodesis was 10.6 days and mean time to fistula resolution after pleurodesis was 1.5 days. Twenty-three (85%) patients had persistent pleuropulmonary air leak closed with the above procedure. CONCLUSION: Treating persistent pleuropulmonary air leak with autologous blood is promising, but further studies are required to quantify its real effectiveness. PMID- 19556744 TI - Applications of nucleic acid chaperone activity of CspA and its homologues. AB - In Escherichia coli, the cold shock response is exerted upon temperature change from 37 to 15 degrees C and is characterized by induction of several cold shock proteins including its major cold shock protein, CspA. E. coli CspA family consists of nine members, CspA to CspI. CspA and some of its homologues play a critical role in cold acclimation of cells as RNA chaperones by destabilizing secondary structures in RNAs. Here, we showed that the nucleic acid melting activity of Csp proteins can be used to facilitate reactions, such as RT-PCR or RNA cleavage reactions by endoribonucleases, which are hindered by presence of secondary structures in the DNA/RNA substrate used. The low substrate specificity of Csps together with their compatibility with various enzymes and their stability and activity over a broad temperature range makes them ideal candidates to be used for a variety of processes. PMID- 19556746 TI - Interaction between plants and bacteria: glucosinolates and phyllospheric colonization of cruciferous vegetables by Enterobacter radicincitans DSM 16656. AB - For determining interactive plant-bacterial effects between glucosinolates and phyllospheric colonization by a plant growth-promoting strain, Enterobacter radicincitans DSM 16656, in cruciferous vegetables, the extent of bacterial colonization was assessed in 5 cruciferous vegetables (Brassica juncea, Brassica campestris, Brassica oleracea var. capitata, Brassica rapa var. alboglabra, Nasturtium officinale) using a species-specific TaqMan probe and quantitative real-time PCR. Colonization ability of inoculated E. radicincitans in the phyllosphere of these species varied from inability to colonize B. rapa up to a very good colonization rate of B. campestris. In addition to morphological factors and other plant compounds, the colonization rate was affected by different individual aromatic and aliphatic glucosinolates and their concentration, revealing that both plant pathogens and plant growth-promoting bacteria were affected by glucosinolates in their colonization behavior. In contrast, after E. radicincitans inoculation neither the total nor the individual glucosinolate concentrations in the phyllosphere of the 5 cruciferous species were affected, indicating that the nonpathogenic E. radicincitans might cause only poor cell damage by metabolizing plant cell components and does not induce a plant defense response and thus subsequently an increased glucosinolate concentration in the phyllosphere. Moreover, E. radicincitans induced no stimulation of indole glucosinolate biosynthesis by additional bacterial auxin supply. PMID- 19556745 TI - Effect of Lactococcus lactis CLFP 100 and Leuconostoc mesenteroides CLFP 196 on Aeromonas salmonicida Infection in brown trout (Salmo trutta). AB - Aeromonas salmonicida is the etiological agent of furunculosis in salmonid fish. This pathogen is important from an epizootic perspective because fish surviving an outbreak can remain lifelong asymptomatic carriers, serving as reservoirs of infection. As a result, the early detection and the control of infection are essential to prevent the spread of new furunculosis outbreaks. We have thus analyzed the effect of probiotic administration on the incidence of A. salmonicida in brown trout (Salmo trutta), that were subjected to temperature stress. Treatment with probiotic strains (Lactococcus lactis CLFP 100 and Leuconostoc mesenteroides CLFP 196) resulted in a higher survival rate after challenge, activation of phagocytic cells in the head kidney, and a lower rate of pathogen proliferation in the intestine as determined by real-time PCR. PMID- 19556747 TI - Sequencing and expression of the xylanase gene 2 from Trichoderma reesei Rut C-30 and characterization of the recombinant enzyme and its activity on xylan. AB - The Xyn2 gene, which encodes endo-beta-1,4-xylanase2, in Trichoderma reesei Rut C 30 was amplified by PCR from first-strand cDNA synthesized on mRNA isolated from the fungus. The nucleotide sequence of the cDNA fragment was verified to encode 190-amino-acid residues of a protein with a calculated molecular mass of 21 kDa. The cDNA was cloned into pET30alpha expression vector and subsequently expressed in Escherichia coli under the control of strong bacteriophage T7 transcription and translation signals. The enzyme activity assay verified the recombinant protein as xylanase. The isoelectric point and highest activity were 7.5 and 1,600 U/mg, respectively. Like with the T. reesei Xyn2, the highest activity of the recombinant Xyn2 was at 50 degrees C. However, the recombinant enzyme had an improved thermostability and more than 65% of its activity retained after 30 min incubation at 60 degrees C. In addition, the recombinant Xyn2 was active over the range of pH 3.5-7.5 with maximum activity at pH 5.0. Using birchwood xylan, the determined apparent K(m) and k(cat) values were 0.15 mg/ml and 119.7 s(-1), respectively. The enzyme was highly specific towards xylans and exhibited very low activity towards cellulosic substrates. Analysis of the products from birchwood xylan degradation confirmed that the enzyme was an endo-xylanase with xylobiose and xylose as the main degradation products. These properties should make the enzyme a suitable applicant in various industrial applications. PMID- 19556748 TI - Developing the blueprint for a genetic testing registry. AB - While the number of genetic tests continues to grow, publicly accessible information about the analytic and clinical validity of such tests is lagging. Information gaps impede informed decision making by health care providers and patients. Enhancing the transparency of information about what tests are being offered, for which indications tests are being offered, and the analytic and clinical validity of tests is a key prerequisite to ensuring test quality. A recent government recommendation for a mandatory genetic test registry has received wide stakeholder support but leaves many practical questions unanswered. We propose a 'blueprint' for the creation of a genetic test registry in order to expedite its implementation. We describe the goals of a registry, propose criteria for the inclusion of registrants and tests in the registry, and define the categories of information that should be included for such tests. We discuss the sources of legal authority that empower the government to mandate that a registry be established and identify the federal agencies with the relevant expertise and resources to do so. We conclude that establishing a registry is a critical first step in the development of a more transparent, quality-centered system of oversight that will better inform and protect the public. PMID- 19556749 TI - Population screening for genetic disorders in the 21st century: evidence, economics, and ethics. AB - BACKGROUND: Proposals for population screening for genetic diseases require careful scrutiny by decision makers because of the potential for harms and the need to demonstrate benefits commensurate with the opportunity cost of resources expended. METHODS: We review current evidence-based processes used in the United States, the United Kingdom, and the Netherlands to assess genetic screening programs, including newborn screening programs, carrier screening, and organized cascade testing of relatives of patients with genetic syndromes. In particular, we address critical evidentiary, economic, and ethical issues that arise in the appraisal of screening tests offered to the population. Specific case studies include newborn screening for congenital adrenal hyperplasia and cystic fibrosis and adult screening for hereditary hemochromatosis. RESULTS: Organizations and countries often reach different conclusions about the suitability of screening tests for implementation on a population basis. Deciding when and how to introduce pilot screening programs is challenging. In certain cases, e.g., hereditary hemochromatosis, a consensus does not support general screening although cascade screening may be cost-effective. CONCLUSION: Genetic screening policies have often been determined by technological capability, advocacy, and medical opinion rather than through a rigorous evidence-based review process. Decision making should take into account principles of ethics and opportunity costs. PMID- 19556750 TI - What can interest tell us about uptake of genetic testing? Intention and behavior amongst smokers related to patients with lung cancer. AB - BACKGROUND: Much of the research examining psychosocial aspects of genetic testing has used hypothetical scenarios, based on the largely untested assumption that hypothetical genetic testing intentions are good proxies for behavior. We tested whether hypothetical interest predicts uptake of genetic testing and whether factors that predict interest also predict uptake. METHODS: Participants (n = 116) were smokers and related to patients with lung cancer, who completed a telephone survey. Interest in genetic testing for lung cancer risk was indicated by responding 'definitely would' to a Likert-style question. Internet-delivered genetic testing for lung cancer risk was then offered. Uptake was indicated by requesting the test and receiving the result. RESULTS: 63% of participants said they 'definitely would' take the genetic test; uptake was 38%. Participants who said they 'definitely would' take the test were more likely than others to take the offered test (45% vs. 26%, p = 0.035). Interest was associated with attitudes towards genetic testing and motivation to quit smoking. Uptake was associated with motivation, prior awareness of genetic testing, and daily Internet use. CONCLUSION: Hypothetical interest only modestly predicts uptake of genetic testing. Interest in genetic testing likely reflects generally positive attitudes that are not good predictors of the choices individuals subsequently make. PMID- 19556751 TI - A pilot beta-thalassaemia screening program in the Albanian population for a health planning program. AB - In Albania, no definite national screening programme of beta-thalassaemia has yet been developed for carrier detection. Only limited information about the occurrence and the types of haemoglobin abnormalities is available. Thus, an educational and screening programme was carried out in one high school with a total of 217 young students from the coastal province of Lushnja in Albania. The pilot programme included a systematic sampling of whole saliva, DNA genomic extraction and the determination of defective beta-thalassaemia genes by reverse dot-blot hybridization with 22 probes specific for the Mediterranean populations.Of the 201 subjects tested, 17 (8.4%) students turned out to be carriers of beta-thalassaemia mutations and haemoglobin variants. The most common mutation is HbS (c.20A-->T) with a frequency of 3.2%, followed by IVS-I-110 (G- >A) (c.93-21G-->A) substitution identified in 4 out of 402 chromosomes (1%). In the province of Lushnja, the frequency of beta-thalassaemia carriers was high. As expected, the results show that identified mutations in this population are similar to those found in the east Mediterranean area, suggesting the same origin for mutant alleles during migratory streams. Implementation of a routine carrier screening programme is significantly facilitated by the presence of only two mutations and would be a wise approach to prevent beta-thalassaemia in the region. PMID- 19556753 TI - A new perspective on therapeutic inhibition of advanced glycation in diabetic microvascular complications: common downstream endpoints achieved through disparate therapeutic approaches? AB - A commonality among the chemically disparate compounds that inhibit the formation and accumulation of advanced glycation end products (AGEs) or their signalling pathways is their end organ protection in experimental models of diabetes complications. Although this group of therapeutics are structurally and functionally distinct with numerous mechanisms of action, the most important factor governing their therapeutic capability is clearly their ability to alleviate the tissue burden of advanced glycation, rather than the biochemical mechanism by which this is achieved. However, it remains to be determined if it is the reduction in tissue AGE levels per se or inhibition of downstream signal pathways which is ultimately required for end organ protection. For example, a number of these agents stimulate antioxidant defences, modify lipid profiles and inhibit low-grade inflammation. These novel actions emphasise the importance of further examination of the advanced glycation pathway and in particular the diverse action of these agents in ameliorating the development of diabetic complications such as nephropathy. PMID- 19556752 TI - Autologous peripheral blood stem cell transplantation to treat CHOP-refractory aggressive subcutaneous panniculitis-like T cell lymphoma. PMID- 19556754 TI - Current status and perspective of the dialysis fluid delivery system in relation to dialysis fluid purification. Preface. PMID- 19556755 TI - Worldwide guidelines for the preparation and quality management of dialysis fluid and their implementation. AB - Quality standards for dialysis water have existed for more than 25 years. Current standards generally agree concerning the maximum allowable levels of chemical contaminants; however, significant differences exist concerning the maximum allowable levels of microbiological contaminants and the methods used to measure them. While quality standards for dialysis water are common, there are relatively few standards or recommendations for dialysis fluid quality and these also differ markedly in the maximum allowable level for microbiological contaminants. Compliance with quality standards for dialysis water and dialysis fluid appears to have improved over the past 20 years, although the actual extent of compliance is difficult to assess. A universal standard for fluid quality might be of benefit to the wider dialysis community; however, progress towards that goal will depend on resolution of important issues, including how the standard is to be applied, if it should be limited to substances with documented toxicity for hemodialysis patients, and how to address microbiological contaminants. PMID- 19556756 TI - The new standard of fluids for hemodialysis in Japan. AB - The standard of fluids for hemodialysis is being evaluated by the International Organization for Standardization (ISO), and will be decided within a few years. In 2008, the Japanese Society for Dialysis Therapy (JSDT) proposed the standard of fluids for hemodialysis by taking the draft ISO standard into consideration and the circumstances in Japan. It was characteristically a standard for Japan, where the central dialysis fluid delivery system (CDDS) is routinely used. In addition, the therapeutic application of each dialysis fluid is clarified. Since high-performance dialyzers are frequently employed in Japan, the standard recommends that ultrapure dialysis fluid be used for all dialysis modalities at all dialysis facilities. It also recommends that the dialysis equipment safety management committee at each facility validate the microbiological qualities of online-prepared substitution fluid, making the responsibility of the dialysis facility clear. This standard is more rigid than those of other countries, and is expected to contribute to improvements in the survival outcome of dialysis patients. PMID- 19556757 TI - Bacteriological water quality in the central dialysis fluid delivery system from the survey of the Japanese Society for Dialysis Therapy. AB - The Japanese Society for Dialysis Therapy (JSDT) surveyed all dialysis facilities for bacteriological quality of dialysis fluid and quality controls for dialysis fluid in 2006 and 2007. The JSDT collected the data for endotoxin (ET) levels, bacterial count and usage of ET retentive filters (ETRF). The JSDT standard for ET level in dialysis fluid (<0.050 EU/ml) was achieved in 89.0% in 2006 and in 93.6% in 2007. The JSDT standard for bacterial cell counts in dialysis fluid (<100 cfu/ml) was achieved in 96.9% in 2006 and in 97.4% in 2007. The central dialysis fluid delivery system (CDDS) is a unique system developed in Japan which has easy handling for daily maintenance of delivery systems, but it has been pointed out that CDDS has a weak point for the protection of biofilms. However, the bacteriological water qualities of dialysis fluid in CDDS were proven to be extremely high in most Japanese dialysis facilities by JSDT surveys. Bacteriological water quality has a strong impact on the patient outcome. The acceptable level of ET of dialysis fluid should be <0.1 EU/ml based on the results of JSDT survey. The excellent water quality in CDDS might be one of the important factors which help good patient survival in chronic dialysis in Japan. PMID- 19556758 TI - Purification of dialysis fluid: historical background and perspective. AB - When dialysis became a chronic therapy, certain clinical symptoms could be connected to the fluid quality and some form of water treatment had to be introduced. The required equipment was empirically developed and consisted of sedimentation filters, carbon filters and softeners. In the mid-1970s the toxic effect of aluminum accumulation was discovered and led to the introduction of reverse osmosis modules. When these components - prefilters, softeners and RO modules - are properly maintained, they produce water of a quality that should meet modern standards. However, the water quality could be ruined by bacterial contamination from the distribution pipes, unless the entire flow path is hygienically designed and frequently disinfected. The quality of the concentrate is also important, especially the bicarbonate component which is prone to bacterial growth. The extent of the microbiological burden in water and dialysis fluid has been brought to the attention of the dialysis community through new and sensitive detection and quantification methods for bacteria and endotoxin. PMID- 19556759 TI - Clinical effect of purification of dialysis fluids, evidence and experience. AB - Microinflammation in renal failure has been the subject of numerous contributions to the literature. The bacterial contamination of dialysate and subsequent transfer of bacterial products onto the blood side is an important cause for microinflammation in hemodialysis patients. It has been suggested that the inflammatory process may not merely be an epiphenomenon but rather a pathogenetic factor in the genesis of atherosclerosis [1,2,3]. Thus, by promoting microinflammation, dialysate contamination may contribute directly to cardiovascular morbidity and mortality. PMID- 19556760 TI - Advances and advantages in recent central dialysis fluid delivery system. AB - The central dialysis fluid delivery system (CDDS) has been working exclusively in Japan since 1970, a fact which proves that CDDS is useful in the multi-patient treatment with in-center regular hemodialysis. It is a cost-effective, labor saving and time-tested system with 40 years of experience. Microbial safety is improved by a refined system design, multiple endotoxin retentive filters (ETRF) and daily overnight disinfection of the entire system. Consequently, the usage of backfiltered dialysate is now accepted by regulatory authorities in some bedside consoles and even more the creation of dialysate is possible for infusion to meet its requirements. In many countries, especially in Asia, the number of end-stage renal disease patients is rapidly increasing. CDDS will contribute to such emerging situations with its easy handling and economical advantages. PMID- 19556761 TI - Current situation of endotoxin retentive filter. AB - Most Japanese facilities use central dialysis fluid delivery systems (CDDS). In addition, since high-performance dialysis membranes are frequently used in Japan, the standard recommends that ultrapure dialysis fluid (UPD) should be used for all dialysis modalities at all dialysis facilities. RO produces and endotoxin retentive filters (ETRFs) occupy the most important position for UPD in this CDDS process. But ETRF is not accepted as medical equipment now in Japan. Since the logarithm reduction value (LRV) of ETRFs widely used today may be a desirable 3 for endotoxin (ET) and 7 for bacteria, UPD could be theoretically obtained through the use of ETRFs. However, we have no data of the LRV for ET and bacteria with a both new and used ETRF now. So we must study whether ETRF has this performance by doing an ET and bacterial challenge test in the near future. Also, ETRF will have to be accepted as medical equipment indicating the safety of CDDS. PMID- 19556763 TI - Guidance of technical management of dialysis water and dialysis fluid for the Japan Association for Clinical Engineering Technologists. AB - There has been remarkable medical and technological progress in Japanese dialysis therapy where more than 270 thousand patients had been treated with dialysis by the end of 2007. Clinical engineering technologists have played an important role not only in the safety treatment but also in the technological development of dialysis therapy. It is very important to supply pure dialysis fluid for both the efficacy and the safety of hemodialysis in which high permeable dialysis membranes are used. The Japanese Society for Dialysis Therapy recently issued the standard for bacterial management of fluids for hemodialysis and related therapies according to the International Organization for Standardization (ISO)/DIS 23500. In order to achieve the standard, the management of dialysis water treatment is important as well as the role of clinical engineering technologists in daily dialysis practice. Purification is defined as no contamination by chemical substances and/or microorganisms and its components. The purification consists of the design and the system structure of the water treatment equipment and dialysis fluid-supplying equipment, and the operation and management of the equipment. The guideline aims to show the minimum standard and the management method of the water treatment system and dialysis fluid-supplying equipment in order to perform hemodialysis safely. They should outline safer dialysis by the management of purification of dialysis fluid. PMID- 19556762 TI - Standardization of water purification in the central dialysis fluid delivery system: validation and parametric method. AB - The central dialysis fluid delivery system (CDDS) has been mainly used for hemodialysis therapy in Japan. Validation and a parametric method are necessary for the quality control of dialysis fluid in CDDS. Validation is a concept for the assurance of system compatibility and product quality, and is defined as follows: the manufacturing and quality control methods including the system design and equipment of the manufacturing facility, manufacturing procedure and processes. Confirmed results must be kept within acceptable limits and they must be documented in a record. Important parameters for validating CDDS include: (1) setting the sterilized area; (2) decision of sterilization level; (3) confirmation of the maximum bio-burden; (4) performance of endotoxin retentive filter and reverse osmosis (RO) module, and (5) checkpoints of purity of dialysis water in the system. Taking the concept of validation and a parametric method in the management of CDDS into consideration enables the supply the purified dialysis fluid or the online prepared substitution fluid that meet the 2008 standards of the Japanese Society for Dialysis Therapy. PMID- 19556764 TI - Central online hemodiafiltration in Japan: management of water quality and practice. AB - Hemodiafiltration (HDF) includes a variety of technologies and preparation of ultrapure dialysis fluid has made it possible to perform online HDF and its extensive alternatives. According to current statistics, 5.8% of ESRD patients are treated with HDF in Japan. The majority of these HDF treatments are performed using the central dialysis fluid delivery system (CDDS), this is because most Japanese clinicians and researchers consider that with CDDS it is easier to prepare substitution fluid; moreover, CDDS has economical advantages against single-patient dialysis machine (SPDM)-based counterparts. The water quality at each patient station (dialysis console) is regularly validated by bacterial culture (colony-forming units) and by measuring endotoxin concentration (ET). Since ET measurement takes much less time than bacterial culture, ET is often used as an indicator to verify the water quality for online use. Dialysis fluid with ET below the detection level (usually <0.001 EU/ml) is used for online substitution. In CDDS online HDF, since dialysis clinics must prepare not only the dialysis fluid but also the substitution fluid, they need to satisfy almost the same requirements as pharmaceutical water treatment factories do. The Japanese Society for Dialysis Therapy (JSDT) together with the Japanese Society for Hemodiafiltration (JS-HDF) is now preparing guidelines to meet all these necessary requirements on a worldwide basis. PMID- 19556765 TI - Fully automated dialysis system based on the central dialysis fluid delivery system. AB - The fully automated dialysis system (FADS) was developed as an improvement over previous patient monitors used in the treatment of hemodialysis, with the aim of standardizing and promoting labor-saving in such treatment. This system uses backfiltration dialysis fluid to perform priming, blood rinse back and rapid fluid replenishment, and causes guiding of blood into the dialyzer by the drainage pump for ultrafiltration. This requires that the dialysis fluid used be purified to a high level. The central dialysis fluid delivery system (CDDS) combines the process of the creation and supply of dialysis water and dialysis fluid to achieve a level of purity equivalent with ultrapure dialysis fluid. FADS has the further advantages of greater efficiency and streamlined operation, reducing human error and the risk of infection without requiring the storage or disposal of normal saline solution. The simplification of hemodialysis allows for greater frequency of dialysis or extended dialysis, enabling treatment to be provided in line with the patient's particular situation. FADS thus markedly improves the reliability, safety and standardization of dialysis procedures while ensuring labor-saving in these procedures, making it of particular utility for institutions dealing with dialysis on a large scale. PMID- 19556766 TI - Purification of dialysis water in the central dialysis fluid delivery system in Japan: a prospective observation study. AB - Whereas the main measure of dialysis fluid purity is endotoxin (ET) activity in Japan, it is the viability count in Western countries. Because of this difference, little information is available concerning dialysis fluid purity determined in terms of viability count in Japan. Under these circumstances a fact finding investigation was planned and conducted concerning dialysis fluid purity to demonstrate the effectiveness of dialysis fluid purification measures. 93 medical institutions are equipped with the central dialysis fluid delivery system (CDDS) unique to Japan. Almost all medical institutions surveyed have achieved the purification level of ultrapure dialysis fluid after ETRF, but the methods of ETRF use and management widely vary with each institution so that early validation of the methods of evaluation of ET inhibition and system management is in urgent need. It is also important that simple universal microbial monitoring and purification procedures be diffused far and wide as suggested by the Purification Guidelines proposed by us. PMID- 19556767 TI - Current status of central concentrate delivery system for hemodialysis in Korea. AB - So far, less than 10% of hemodialysis (HD) facilities in Korea have adopted a kind of central delivery system (CDS) for dialysates, the so-called Korean central concentrate delivery system (CCDS). In our CCDS, all concentrate solutions of the acid and bicarbonate dry powder mixed with the treated reverse osmosis (RO) water in the mixing tank and stored in the holding tank are centrally delivered to individual HD machines via two streams along with a one further stream for dilution RO water. This mode of Korean CCDS is different from the central dialysate delivery system (CDDS), which uses already fully proportioned acid and bicarbonate concentrates with dilution RO water delivering it via a single stream. At present, the most popular CCDS in Korea is NephroMix Premium, which is an all-in-one system that combines the mixing tank, the holding tank, and the control functions into one unified case. Moreover, all CCDS data can be monitored in real-time through a user-friendly touch-screen panel, and stored on a desktop PC linked to the manufacturer's main office. For the disinfection and sterility of NephroMix Premium, ozone is used besides a three step filtering system for removing endotoxin and microbials. Compared to the conventional system in Korea with individual concentrate delivery to single patient dialysis machines, the Korean-style CCDS seems to be superb in the way of convenience, environmental friendliness, and economy, and also needs less space in the dialysis unit. However, full proof of its safety and effectiveness is required for further verification in the future. PMID- 19556768 TI - Poststroke depression: prevalence and determinants in Brazilian stroke patients. AB - BACKGROUND: Poststroke depression (PSD) is one of the most important long-term adverse psychosocial consequences in stroke survivors. Our objective was to assess the prevalence of PSD in Brazilian stroke patients and identify significant associated factors. METHODS: A cross-sectional study of stroke patients consecutively admitted for rehabilitation was conducted. The patients were evaluated by means of the NIH Stroke Scale, Mini-Mental State Examination, Barthel Index, Lawton Scale, modified Rankin Scale, Hospital Anxiety and Depression Scale (HADS), Geriatric Depression Scale (GDS) and MOS-Short Form 36. Patients with a HADS-depression subscale score > or = 11 and/or GDS score > or = 8 were classified as depressed. RESULTS: Three hundred stroke survivors were assessed (mean age: 56.3 years; 51.7% males). Half (46.7%) of the stroke patients had an m-RS score < or = 2. The proportion of stroke patients who scored > or = 11 points on the HADS-depression and HADS-anxiety subscales were 19.2 and 23.7%, respectively. One third (29.7%) had a GDS mean score > or = 8. The GDS scores significantly correlated (p < 0.0001) with the HADS-depression (r = 0.51) and HADS-anxiety subscales (r = 0.54). The prevalence of mood disorders was significantly higher in females than in males (24.8 vs. 14.2%; x(2), p = 0.03). PSD was significantly associated (p < 0.0001) with work status (housewife), education level, lower social and cognitive functioning, dependence in the instrumental activities of daily living and presence of diabetes in the multivariable regression analysis (R adjusted = 0.32). CONCLUSIONS: PSD was highly prevalent in the chronic phase of stroke. Early detection and recognition of associated risk factors is important to treat and prevent PSD in a rehabilitation setting. PMID- 19556769 TI - Endothelial function in lacunar infarction: a comparison of lacunar infarction, cerebral atherosclerosis and control group. AB - BACKGROUND: There are conflicting evidences on endothelial function in lacunar infarction. This may be attributed to the effects of risk factors on the vascular smooth muscle. To test endothelial function only in patients with lacunar infarction, we evaluated the endothelium-dependent and -independent vasodilatation of the brachial artery. METHODS: We enrolled consecutive patients with lacunar infarction defined by clinical characteristics and MRI findings. The control group included age- and sex-matched patients with hypertension who do not have any history of clinical stroke, coronary artery disease or peripheral vascular disease. Endothelial function was evaluated using flow-mediated dilatation (FMD) and nitrogen-mediated dilatation (NMD) of the brachial artery. FMD and NMD were examined by an experienced vascular sonographer using a high resolution ultrasound. Intracranial stenosis was defined as flow gap or >50% reduction in vessel diameter on MRA. RESULTS: FMD was 6.6 +/- 4.5% in the lacunar infarction group and 12.2 +/- 4.6% in the control group (p = 0.000). NMD was 14.3 +/- 4.9% in the lacunar infarction group and 13.8 +/- 4.9% in the control group (p = 0.37). FMD in patients with lacunar infarction and intracranial arterial stenosis was 6.4 +/- 3.9%, and FMD in patients with lacunar infarction was only 6.9 +/- 5.5%. In the control group, it was 12.2 +/- 4.6%. CONCLUSION: FMD was low in patients with lacunar infarction. NMD was similar between the lacunar infarction group and the control group. These results are suggestive of pure endothelial dysfunction in lacunar infarction. Endothelial dysfunction was as severe in lacunar infarction as in intracranial arterial stenosis. PMID- 19556770 TI - An observational study of acute stroke care in four countries: the European registers of stroke study. AB - BACKGROUND: The European Registers Of Stroke (EROS) project aimed to assess outcomes of stroke care across Europe, relating these to both case mix information from disease-specific population registers and the quality of stroke care provided at each centre. This included comparing information on quality of care with direct observation of the stroke care process in 4 centres. METHODS: Direct non-participant observational methods were used on a purposive sample of first-stroke patients admitted within the past 14 days to an acute-stroke unit or ward that admits stroke patients in 4 urban hospital sites in London, UK, Dijon, France, Kaunas, Lithuania, and St. Petersburg, Russia. We recorded patient characteristics with levels of contact with multi-disciplinary team (MDT) members and contact with families and mobilization to build a collection of 'snapshots' of stroke care throughout the patients' day. One independent observer undertook all observations over 1 day. RESULTS: We observed differences between centres in the proportion of observations where patients were alone (lowest proportion in London, highest proportion in St. Petersburg) (p > 0.001), where patients had contact with MDT members (p > 0.001) and family, and where patients were out of bed/mobile (p > 0.001) (both with highest proportion in London, lowest proportion in St. Petersburg). CONCLUSIONS: Higher levels of contact with the MDT, family contact and mobilization were observed in the Western European centres than the Eastern European and Russian centres. Differences in case mix may explain some, but not all, of these differences. Direct observation has some limitations; however, it could be developed in future studies to help identify other key aspects of effective stroke care. PMID- 19556772 TI - Chromosome structure and function. Preface. PMID- 19556773 TI - Telomere distribution and dynamics in somatic and meiotic nuclei of Arabidopsis thaliana. AB - The ends of linear eukaryotic chromosomes are protected by the telomere, a specialised nucleoprotein complex. The primary role of the telomere is to protect the chromosome ends from being degraded or recognised and processed as double strand breaks. Additionally, the telomeres are involved in interphase chromosome organisation and also in chromosome pairing, synapsis and movement during meiotic prophase. The main emphasis of this review is concerned with the distribution and dynamics of the telomeres in the somatic cell and meiocytes of plants, focusing on the model plant Arabidopsis thaliana. In Arabidopsis the telomeres are organised around the nucleolus in both the somatic and meiotic interphase. One of the outstanding questions in meiosis is how homologous chromosomes pair (align) and synapse during meiotic prophase. Recent attention has been paid to the bouquet formation, a nearly universal event, during which the telomeres cluster on the nuclear membrane in early prophase. It has been suggested that because the telomeres are in close proximity this would enhance their pairing and subsequent synapsis of the homologues. In Arabidopsis we observe that the telomeres are paired homologously in early meiosis whilst still arranged around the nucleolus. They are moved to the nuclear membrane preceding synapsis and reveal only a loose clustering, which may represent a transient bouquet. On completion of synapsis the paired telomeres are dispersed and remain attached to the nuclear membrane until diplotene when they dissociate from the nuclear membrane. We also discuss the prospects for live imaging of the telomeres in Arabidopsis. PMID- 19556771 TI - Association of white-matter lesions with brain atrophy markers: the three-city Dijon MRI study. AB - BACKGROUND: Brain atrophy and white-matter lesions (WML) are common features at cerebral MRI of both normal and demented elderly people. In a population-based study of 1,792 elderly subjects aged 65-80 years, free of dementia, who had a cerebral MRI at entry, we investigated the relationship between WML volume and brain atrophy markers estimated by hippocampal, gray matter (GM) and cerebrospinal fluid (CSF) volumes. METHODS: An automated algorithm of detection and quantification of WML was developed, and voxel-based morphometry methods were used to estimate GM, CSF and hippocampal volumes. To evaluate the relation between those volumes and WML load, we used analysis of covariance and multiple linear regression models adjusting for potential confounders and total intracranial volumes. RESULTS: Age was highly correlated with WML load and all brain atrophy markers. Total WML volume was negatively associated with both GM (beta = -0.03, p < 0.0001) and hippocampal volumes (beta = -0.75, p = 0.0009) and positively with CSF volumes (beta = 0.008, p = 0.02) after controlling for sex, age, education level, hypertension and apolipoprotein E genotype. Evidence for a relationship between brain atrophy markers and WML was stronger for periventricular WML. We found that the relationship between WML and hippocampal volumes was independent of other brain tissue volumes. CONCLUSION: These results suggest that, in the brain of nondemented elderly subjects, degenerative processes and vascular changes co-occur and are related independently of vascular risk factors. PMID- 19556774 TI - SMC proteins and their multiple functions in higher plants. AB - In plants as in other eukaryotes, SMC (Structural Maintenance of Chromosome) protein complexes and proteins interacting with them are essential for sister chromatid cohesion, chromosome condensation, DNA repair and recombination. The presence of paralogous genes for various components of the different SMC complexes allowed diversification of their biological functions during evolution of higher plants. Here I summarize the multiple functions of plant SMC complexes and some of the particularities these proteins show in comparison to those of other organisms. PMID- 19556775 TI - Structural analyses of chromosomes and their constituent proteins. AB - The importance of chromosome structural study is first described. Then an overview of historical imaging methods that enable us to quantitatively understand chromosome images and structure is given with special reference to the identification of small plant chromosomes and development of their quantitative chromosome maps. A three-dimensional understanding of chromosome distribution within a nucleus answers why the gene-rich regions localize at both ends of chromosomes, especially in the case of species with Rabl orientation. Not only imaging methods but also proteomic approaches are effective in understanding chromosome structure. Over 200 proteins have been identified by proteome analysis of human metaphase chromosomes, and are categorized into four distinct groups according to their nature and localization on chromosomes. These are chromosome coating proteins (CCPs), chromosome peripheral proteins (CPPs), chromosome structural proteins (CSPs), and chromosome fibrous proteins (CFPs). A chromosome four layer model has been developed accordingly. Case studies on individual identified proteins are further described, and the functional similarities of CPPs are exemplified. In addition the controversial roles of CSPs (topoisomerase and condensin), especially for development of higher-order chromosome structure, are discussed. Finally, it is concluded that further advances in chromosome research are necessary to solve an enigma lasting nearly two centuries, that is, why chromosomes retain the same shape in plants and animals. PMID- 19556777 TI - Chromosome centromeres: structural and analytical investigations with high resolution scanning electron microscopy in combination with focused ion beam milling. AB - Whole mount mitotic metaphase chromosomes of different plants and animals were investigated with high resolution field emission scanning electron microscopy (FESEM) to study the ultrastructural organization of centromeres, including metacentric, acrocentric, telocentric, and holocentric chromosome variants. It could be shown that, in general, primary constrictions have distinctive ultrastructural features characterized by parallel matrix fibrils and fewer smaller chromomeres. Exposure of these structures depends on cell cycle synchronization prior to chromosome isolation, chromosome size, and chromosome isolation technique. Chromosomes without primary constrictions, small chromosomes, and holocentric chromosomes do not exhibit distinct ultrastructural elements that could be directly correlated to centromere function. Putative spindle structures, although rarely observed, spread over the primary constriction to the bordering pericentric regions. Analytical FESEM techniques, including specific DNA staining with Pt blue, staining of protein as a substance class with silver-colloid, and artificial loosening of fixed chromosomes with proteinase K, were applied, showing that centromere variants and ultrastructural elements in the centromere differ in DNA and protein distribution. Immunogold localization allowed high-resolution comparison between chromosomes with different centromere orientations of the distribution of centromere-related histone variants, phosphorylated histone H3 (ser10), and CENH3. A novel application of FESEM combined with focused ion beam milling (FIB) provided new insights into the spatial distribution of these histone variants in barley chromosomes. PMID- 19556778 TI - Avian lampbrush chromosomes: a powerful tool for exploration of genome expression. AB - Lampbrush chromosomes (LBCs) are highly extended bivalents that function in the growing oocytes of many animals. Due to their distinctive chromomere-loop organization and intense transcriptional activity of lateral loops the LBCs, mainly amphibian ones, have served as a powerful system for exploring the general principles of chromosome organization and function. The exploitation of avian LBCs has considerably broadened the opportunities for comparative genome research and for cytogenetic analysis of domestic species. In this review we highlight the advantages of avian LBCs for research in different areas including integration of genome organization studies with studies on gene activity in vivo, analysis of co transcriptional events occurring on nascent transcripts and investigation of chromosome-associated intranuclear domains. Recent findings concerning the organization of transcriptionally active and silent chromatin together with involvement of cohesin and condensin complexes into maintenance of structural integrity of LBCs are presented. The biological significance of the LBC phenomenon is discussed. The intensive transcription on LBCs shows some specific features: very long transcription units, deregulated termination, and transcription of non-coding satellite repeats. Here, based on the modern view on a role of RNA interference machinery in regulation of genome expression, we suggest a mechanism of initiation of satellite DNA transcription and offer a novel interpretation of the 'classical' hypothesis that sought to explain the significance of widespread transcription during oocyte growth. PMID- 19556776 TI - Identification of a maize neocentromere in an oat-maize addition line. AB - We report a neocentromere event on maize chromosome 3 that occurred due to chromosome breakage. The neocentromere lies on a fragment of the short arm that lacks the primary centromere DNA elements, CentC and CRM. It is transmitted in the genomic background of oat via a new centromere (and kinetochore), as shown by immunolocalization of the oat CENH3 protein. Despite normal transmission of the maize fragment in most progeny, neocentromeres appear to vary in size within the same tissue, as shown by fluorescent measurements. A secondary truncation in one line lowered mitotic transmission to 3% and precipitously reduced the size of the chromosome. The results support the view that neocentromere formation is generally associated with major genomic disturbances such as wide species crosses or deletion of an existing centromere. The data further suggest that new centromeres may undergo a period of instability that is corrected over a period of several generations. PMID- 19556779 TI - Transcription of pericentromeric heterochromatin in beetles--satellite DNAs as active regulatory elements. AB - Tenebrionid beetles from the genus Palorus (Coleoptera) have a significant amount of heterochromatin in pericentromeric regions of all chromosomes. The major DNA component of pericentromeric heterochromatin is a highly abundant satellite DNA. Analysis of transcription of a major satellite DNA PSUB from species Palorus subdepressus reveals a constitutive level of expression similar in all 3 developmental stages: larvae, pupae and adults, corresponding to 0.01% of total RNA. Transcription proceeds from both DNA strands in equal amounts resulting in long heterogeneous size transcripts ranging in size from 500 bp to more than 5 kb. Although equal transcription from both DNA strands could potentially activate the RNA interference (RNAi) pathway processing of long PSUB transcripts into small interfering RNAs (siRNA) was not detected. Transcripts preferentially remain in the nucleus and 90% of them are not polyadenylated. Transcription initiation sites, mapped by primer extension, are located within PSUB satellite monomers as well as motifs characteristic for RNA polymerases II and III. Putative RNA polymerase II promoter, predicted by computational approach, shares a 65% sequence similarity to the Pol II promoter mapped previously in PRAT satellite DNA, a major satellite of related species P. ratzeburgii. Results give strong indications that Palorus satellite DNAs are transcribed as autonomous transcription units from their own promoters that reside within satellite sequences. Long satellite DNA transcripts remain mostly in the nucleus and are proposed to play a structural role in the organization of pericentromeric heterochromatin. PMID- 19556780 TI - Satellite DNA spatial localization and transcriptional activity in mouse embryonic E-14 and IOUD2 stem cells. AB - The formation of heterochromatin begins in the differentiating cells. The aim of this work was to study changes of satellite DNA distribution, transcriptional activity and interaction with certain proteins in mouse embryonic stem cells after induction with retinoic acid. We found that pericentromeric satellites entered chromocenters only some days after induction of E-14 and IOUD2 mouse embryonic stem cells. The redistribution was accompanied by association with HP1a and transcription from pericentromeric (but not centromeric) satellite DNA. RNA was polyadenylated and transcribed from the forward chain. Probes made from the cDNA hybridized to all chromosomes. In differentiating cells, the transcript was found exclusively in chromocenters while in differentiated cultured L929 cells it formed 1-2 large clusters outside chromocenters. Using ChIP and immunostaining, we demonstrated that in induced cells pericentromeric DNA interacted with RNA helicase p68 that was previously shown to be involved in transcription regulation and to be involved in differentiation processes. PMID- 19556781 TI - SAF-A/hnRNP-U localization in interphase and metaphase. AB - SAF-A/hnRNP U is an abundant nuclear protein that interacts specifically with nuclear matrix attachment region DNA (MAR) and RNA as a component of hnRNPs. SAF A/hnRNP U was also shown to specifically bind mouse major satellite DNA (satMa). Antibodies against SAF-A and GFP-fusion constructs were used in the current work in order to trace SAF-A localization. In accordance with its diverse nucleic acid binding specificity, SAF-A was found to be localized in three different domains: outside the chromosomes, on the surface of the chromosome arms (probably MARs), and in the centromere region where it apparently binds specifically to the satMa. GFP-fusion constructs with different SAF-A/hnRNP U domains confirms the functional significance of the protein's functional domains in interphase cells. In telophase cells, the anti-SAF-A antibody signal appeared as a kind of network covering unfolded chromosomes. PMID- 19556782 TI - Interaction study of the male specific lethal (MSL) complex and trans-acting dosage effects in metafemales of Drosophila melanogaster. AB - The effect of ectopic expression of male specific lethal 2 (msl2) on chromatin modification and gene expression was studied in Drosophila diploid females and metafemales (3X;2A). Results show that ectopic expression of MSL2 in transgenic msl2 females and metafemales sequesters the MOF histone acetylase to the X, which occurs concordantly with an increase of histone acetylation. Gene expression studies indicate that the X-linked genes are not affected by direct targeting of the MSL complex and the resulting increased H4Lys16 acetylation on the X chromosomes, suggesting one function of the MSL complex is to nullify the effect of a high level of histone acetylation. These results are not consistent with the hypothesis that the presence of the MSL complex conditions a two-fold upregulation. Autosomal gene expression is generally decreased in ectopically expressed MSL2 females, which correlates with the reduced autosomal histone acetylation. Metafemales show dosage compensation of X-linked genes with some autosomal reductions in expression. Interestingly, in metafemales with ectopically expressed MSL2, the autosomal expression is returned to a more normal level. There is a lower autosomal level of histone acetylation compared to the normal metafemales, suggesting a nullifying effect on the negative dosage effect of the X chromosome as previously hypothesized to occur in normal males. PMID- 19556783 TI - Extrachromosomal double minutes and chromosomal homogeneously staining regions as probes for chromosome research. AB - Gene amplification in human cancer cells generates two cytogenetically identifiable structures: extrachromosomal double minutes (DMs) and the chromosomal homogeneously staining region (HSR). DMs are composed of autonomously replicating circular DNA of genomic origin, and they tell us about how the extrachromosomal elements may behave in the cells, how they were entrapped by the micronuclei and how they were eliminated from the cells. On the other hand, the episome model predicts that extrachromosomal elements excised from the chromosome arm might generate DMs, and the breakage-fusion-bridge (BFB) cycle model explains the generation of the HSR. In accordance with this, a plasmid bearing a mammalian replication initiation region (IR) and a matrix attachment region (MAR) mimics gene amplification and generates DMs and HSRs de novo. The IR/MAR gene amplification system extends our understanding on the mechanism of gene amplification and the behavior of amplified genes. Furthermore, the system may suggest the way how extrachromosomal elements in general may alter the chromosome architecture and function. PMID- 19556784 TI - Extrachromosomal circular DNA in eukaryotes: possible involvement in the plasticity of tandem repeats. AB - Extrachromosomal circular DNA (eccDNA) is ubiquitous in eukaryotic organisms, and has been noted for more than 3 decades. eccDNA occurs in normal tissues and in cultured cells, is heterogeneous in size, consists of chromosomal sequences and reflects plasticity of the genome. Two-dimensional (2D) gel electrophoresis has been adapted for the detection and characterization of eccDNA. It shows that most eccDNA consists of chromosomal tandem repeats, both coding genes and satellite DNA and is organized as circular multimers of the repeating sequence. 2D gels were unable to detect dispersed repeats within the population of eccDNA. eccDNA, organized as circular multimers, can be formed de novo in Xenopus egg extracts, in the absence of DNA replication. These findings support a mechanism for the formation of eccDNA that involves intra-chromosomal homologous recombination between tandem repeats and looping-out. Furthermore, eccDNA appears to undergo extrachromosomal replication via a rolling circle mechanism. Hence, the formation of eccDNA from arrays of tandem repeats may cause deletions, and the possible re integration of rolling-circle replication products could expand these arrays. This review summarizes recent experimental data which characterizes eccDNA in several organisms using 2D gel electrophoresis, and discusses its possible implications on the dynamics of chromosomal tandem repeats. PMID- 19556785 TI - Role of fluorescence in situ hybridization in sequencing the tomato genome. AB - The tomato (Solanum lycopersicum L.) genome is being sequenced by a consortium of laboratories in 10 countries. Seventy-seven percent of the tomato genome (DNA) is located in repeat-rich, gene-poor, pericentric heterochromatin, while 23% of the genome is located in repeat-poor, gene-rich, distal euchromatin. It is estimated that approximately 90% of tomato's nuclear genes can be characterized by limiting the sequencing effort to euchromatin while avoiding the problems involved in sequencing the repetitive DNA in heterochromatin. Sequencing is being performed on tomato nuclear DNA cloned into bacterial artificial chromosome (BAC) vectors. Fluorescence in situ hybridization (FISH) is used to help direct the sequencing effort by cytologically demonstrating the location of selected BACs on tomato chromosomes. While mitotic metaphase chromosomes are too short and compact for this purpose, long pachytene chromosomes are ideal. BACs localized in euchromatin can be used confidently as anchors for the assembly of BAC contigs that extend through the euchromatic length of each chromosome arm. Another important role for FISH is identification of BACs near telomeres and near borders with pericentric heterochromatin to indicate that sequencing should not extend much further. This role of FISH is enhanced by our ability to estimate base pair distances between localized BACs and these chromosomal features. Finally, it is noteworthy that when BAC-FISH is combined with chromosomal in situ suppression (CISS) hybridization to block repeats and localize single/low copy sequences, the great majority of BACs localize to single sites. This observation is consistent with tomato being an ancient diploid. PMID- 19556786 TI - A technical note on quantum dots for multi-color fluorescence in situ hybridization. AB - Quantum dots (Qdots) are semiconductor nanocrystals, which are photo-stable, show bright fluorescence with narrow, symmetric emission spectra and are available in multiple resolvable colors. We established a FISH protocol for the simultaneous visualization of up to 6 different DNA probes differentially labeled with Qdots and with conventional organic fluorochromes. Using a Leica SP5 laser scanning confocal microscope for image capture, we tested various combinations of hapten labeled probes detected with streptavidin-Qdot525, sheep anti-digoxigenin Qdot605, rat anti-dinitrophenyl-Qdot655 and goat anti-mouse-Qdot655, respectively, together with FITC-dUTP-, Cy3-dUTP- and Texas Red-dUTP-labeled probes. We further demonstrate that Qdots are suitable for imaging of FISH probes using 4Pi microscopy, which promises to push the resolution limits of light microscopy to 100 nanometers or less when applying a deconvolution algorithm, but requires the use of highly photo-stable fluors. PMID- 19556788 TI - In vitro antagonism of rabeprazole and metronidazole upon clinical isolates of Helicobacter pylori. AB - BACKGROUND: Rabeprazole inhibits Helicobacter pylori in vitro. This property could influence the antibacterial activity of metronidazole or amoxicillin. Our goal was to evaluate in vitro the effect of rabeprazole upon the antibacterial activity of metronidazole and amoxicillin. METHODS: The minimal inhibitory concentration (MIC) was determined by agar dilution upon 30 clinical isolates, and the antibacterial effect of the combined drugs was estimated by calculating the fractional inhibitory concentration index (FICI). RESULTS: Nine strains (30%) were resistant to metronidazole (MIC 8-256 mg/l) while only 1 strain was resistant to amoxicillin (MIC 1.0 mg/l). Rabeprazole also inhibited H. pylori (MIC 0.125-1.0 mg/l). The association of metronidazole and rabeprazole showed synergism in 30% of the strains (FICI < or =0.25), but also antagonism in 23.3% of the clinical isolates (FICI > or =4.0). Mostly, amoxicillin-rabeprazole showed an effect of synergism (53.3%). CONCLUSION: The results presented suggest the need to keep in mind that failure in eradication therapies based on metronidazole and rabeprazole could be due to an antagonism of the drugs. PMID- 19556787 TI - Differences in cell wall thickness between resistant and nonresistant strains of Mycobacterium tuberculosis: using transmission electron microscopy. AB - The ultrastructure of the cell wall of extensively drug-resistant (XDR), multidrug-resistant (MDR) and susceptible tuberculosis (TB) bacilli was viewed under transmission electron microscopy (TEM). Under the TEM, marked differences were observed in the thickness of their cell wall: 20.2 +/-1.5 and 17.1 +/-1.03 nm for the XDR and MDR TB bacilli, respectively, and 15.6 +/- 1.3 nm for the susceptible isolates (p < 0.05). In MDR bacilli, thickening of the cell wall was observed in the intermediate electron-transparent layer (ETL) and outer electron opaque layer, whereas in XDR TB cells the basal peptidoglycan layer was denser and almost fused with the ETL. Five to seven percent of XDR TB bacilli had the appearance of the stationary phase with cell wall thickness ranging from 21 to 26 nm (p < 0.001). Information provided in this study is of significant importance in terms of drug selection for effective treatment of resistant strains. PMID- 19556789 TI - Noninvasive evaluation of coronary endothelial function following sirolimus eluting stent implantation by using positron emission tomography. AB - OBJECTIVE: Several invasive studies have reported delayed reendothelialization and endothelial dysfunction following sirolimus-eluting stent (SES) implantation. We evaluated the changes in coronary endothelial function following SES implantation by using a noninvasive method that involved positron emission tomography and cold pressor testing (CPT). METHODS: The study was conducted on 14 lesions on which percutaneous coronary intervention (PCI) was successively performed. The lesions were classified into 2 groups depending on the PCI performed: the conventional PCI group, in which 7 conventional PCIs (plain old balloon angioplasty or bare-metal stents) were performed, and the SES group, in which 7 SESs were implanted. Coronary endothelial function was defined as the percent increase in the myocardial blood flow (MBF) during CPT. RESULTS: The resting MBF in the segments distal to the PCI sites did not differ between the conventional PCI and SES groups; however, the MBF significantly decreased in the SES group during CPT. CONCLUSIONS: These data suggest that SES implantation induces coronary endothelial dysfunction in the segments distal to the PCI sites. PMID- 19556790 TI - Gender difference in the application of reperfusion therapy in patients with acute myocardial infarction. PMID- 19556791 TI - A system of calibrating microtomography for use in caries research. AB - Desktop microcomputed tomography (micro-CT) offers a non-invasive 3-dimensional analysis of structures and their physical properties. To date, the use of micro CT has mostly involved qualitative observations, with the extent of quantitative analysis relying on automated internal calibration by the micro-CT control software. However, the value of such calibration is limited by machine drift. For an accurate quantitative use of micro-CT, it is recognized that external means of calibration are needed. A novel system of calibration standards, also known as 'phantoms', is presented. A range of low mineral concentration phantoms involving triethylene glycol dimethacrylate/glycerolate dimethacrylate resin mixed with commercial pure hydroxyapatite (HAP), from 0.07 to 1.05 g/cm(3), was fabricated. Sintered HAP was impregnated with the same resin, producing phantoms with medium level mineral concentrations up to 1.90 g/cm(3). These phantoms were easy to create, proved accurate and stable with repeated use, and were found to mimic the composite nature of dental enamel and dentine structures under investigation. PMID- 19556792 TI - Solid titration of octacalcium phosphate. AB - Octacalcium phosphate (OCP) is of considerable importance as a precursor in the formation of dental enamel and an intermediate phase in the precipitation of hydroxyapatite (HAp) in bone. However, agreement is poor on the solubility product (pK(sp)), possibly due to the formation of the more stable phase HAp. The system was investigated using solid titration, which has shown reliability in work on HAp and related fluoride minerals, with OCP in 100 mM KCl at 37.0 +/- 0.1 degrees C. The constitution of the end point precipitate was determined by X-ray diffraction and selected-electron area diffraction; the particle morphology and elements present were examined by high-resolution field emission scanning, transmission electron microscopy and energy-dispersive X-ray analysis. The titration curve for OCP was found for pH approximately 3.4-7.4. The precipitate was HAp at pH 3.6 and 4.5; no residual OCP or other phase was detected. Dicalcium phosphate dihydrate (DCPD) was then found to form at pH 3.6 on further addition of OCP titrant after equilibrium had been achieved, possibly due to easier nucleation at lower pH. However, markedly crystalline HAp was formed in equilibrium for OCP titration with HAp seeding, verifying HAp as the more stable phase. A solubility isotherm for OCP was not obtained as HAp appears to be less soluble in the pH range studied. This adds weight to the view that HAp may be the most stable phase of all calcium phosphates, with further doubt being cast on DCPD being the most stable phase below pH 4.2. However, metastable DCPD may form in an Ostwald succession, depending on supersaturation and nucleation conditions. PMID- 19556793 TI - Diagnosis of fat malabsorption by breath tests: just a breeze? PMID- 19556794 TI - Analysis of fat digestive and absorptive function after subtotal gastrectomy by a 13C-labeled mixed triglyceride breath test. AB - BACKGROUND/AIM: In the choice of reconstructions, digestive and absorptive disturbances, resulting in weight loss after subtotal gastrectomy, remain a problem. The aim of this study was to compare fat absorptive function after Billroth I (B-I) and Roux-en-Y (RY) reconstructions after subtotal gastrectomy for gastric cancer. METHODS: A (13)C-labeled mixed triglyceride breath test was performed in 31 patients after subtotal gastrectomy and in 15 healthy volunteers to assess fat digestive and absorptive function. Seventeen B-I reconstructions and 14 RY reconstructions were performed after subtotal gastrectomy. Fat digestive and absorptive function was determined by percent (13)CO(2) cumulative dose at 7 h. Relationship between fat absorptive function and perioperative factors were analyzed. RESULTS: Gender distribution, mean age, pathological staging, level of lymph node dissection, preservative procedure of the vagus nerve and mean follow-up period in the two surgical groups did not differ significantly. Only the type of reconstruction (p = 0.024) was associated with differences in fat digestive and absorptive function by univariate analysis: B-I reconstruction was superior to RY reconstruction. CONCLUSIONS: Fat digestive and absorptive function after B-I reconstruction was superior to that after RY reconstruction, probably because the B-I reconstruction was the procedure that permitted food passage through the duodenum. PMID- 19556795 TI - New prediction rule for mortality in acute mesenteric ischemia. AB - OBJECTIVE: Acute mesenteric ischemia is potentially fatal, but prognostic factors have not yet been established. This study was undertaken to elucidate them. METHODS: This is a retrospective cohort study, consisting of 110 patients who had been treated in the past 5 years, from 26 national hospitals in Japan. RESULTS: The overall in-hospital mortality rate was 51%. Logistic regression analysis demonstrated two independent prognostic factors, electrocardiogram scale with an odds ratio of 1.7 (95% CI 1.2-2.4) and shock index of 11 (95% CI 1.5-80). A stepwise analysis gave a prediction equation for in-hospital mortality (R) using these variables and age score. We further modified this equation to a simpler scoring system (S) using the same variables. Both R and S showed a good discriminatory ability as determined by areas under the receiver-operating characteristic curve (0.83, 95% CI: 0.74-0.91 for R; 0.82, 95% CI 0.74-0.91 for S). The observed mortality rates increased as the R or S increased (19% at R <0.25, 41% at 0.25 < or = R <0.6, 85% at R > or =0.6; 19% at S < or =2, 37% at S of 3 or 4, 91% at S > or =5). CONCLUSION: The new prediction rules can be used at any hospital and may be promising tools for medical decision-making, informed consent and reviewing quality of care. PMID- 19556796 TI - Surgical salvage of bleeding peptic ulcers after failed therapeutic endoscopy. AB - BACKGROUND: The approach to salvage surgery after failed endoscopic therapy for a bleeding peptic ulcer is controversial. We aimed to compare the outcomes of salvage surgery after failed endoscopic therapy for bleeding peptic ulcers over a 10-year period. METHODS: Patients receiving salvage surgery for bleeding peptic ulcers were divided into 2 cohorts, the 1st from 1993 to 1998 and the 2nd from 1999 to 2004. The type of salvage surgery was defined as minimal if ulcer plication or an ulcerectomy was performed, and definitive if the patient received a vagotomy or gastrectomy. RESULTS: One hundred and twenty-three patients received salvage surgery in the 1st cohort, while 42 patients received surgical hemostasis for the bleeding peptic ulcer in the 2nd cohort. Patients in the 2nd cohort consisted of a larger proportion of in-hospital bleeders (cohort 1: 12.2%, cohort 2: 42.9%; p < 0.005) and had a significantly higher proportion of comorbidities. A larger number of patients received minimal surgery in cohort 2 (cohort 1: 42.3%, cohort 2: 73.8%; p < 0.005). CONCLUSIONS: With advances in therapeutic endoscopy, patients who developed failed endoscopic hemostasis are likely to be poor surgical candidates with multiple comorbidities. The approach to salvage surgery has inclined towards minimal surgery to hasten surgical hemostasis among these fragile patients. PMID- 19556797 TI - The effects of L-NG-nitroarginine in a zymosan-induced multiple organ dysfunction syndrome model. AB - BACKGROUND: The aim of this study was to investigate the role of nitric oxide in mesenteric ischemia, organ injury and survival in zymosan-induced multiple organ dysfunction syndrome (MODS) by using the nonselective nitric oxide synthase inhibitor L-N(G)-nitroarginine (L-NNA). METHODS: Swiss albino mice (20-40 g) were used in the study. The animals were randomly divided into four groups. The first group was treated intraperitoneally with saline and served as the sham group for L-NNA. The second group was treated with zymosan (500 mg/kg). The mice in the third and fourth group received L-NNA (20 mg/kg), 1 and 6 h after saline or zymosan administration. Six hours after the administration of zymosan, animals were used for mesenteric arterial blood flow (MABF) measurements and then sacrificed for biochemical and histopathological analyses at the 18th hour. RESULTS: In zymosan-treated animals, MABF was significantly lower than that of solvent saline-treated controls (controls: 4.7 +/- 0.8 ml.min(-1); zymosan: 1.7 +/- 0.7 ml.min(-1), p < 0.05). L-NNA did not prevent zymosan-induced MABF decrease (controls: 4.5 +/- 0.8 ml.min(-1); zymosan: 2.5 +/- 1.4 ml.min(-1), p <0.05). Also treatment with L-NNA has no beneficial effect on survival and organ injury in zymosan-induced MODS. CONCLUSION: In this study, inhibition of both inducible and constitutive nitric oxide synthase by L-NNA did not abolish the harmful effects of zymosan. L-NNA remains an agent without any therapeutic potential in this acute experimental model of MODS. PMID- 19556799 TI - Effect of erythropoietin on oxidative stress and liver injury in experimental obstructive jaundice. AB - BACKGROUND: To evaluate the time-dependent effects and pathophysiological mechanism of erythropoietin (Epo) on oxidative stress and liver injury resulting from obstructive jaundice in common bile duct-ligated rats. METHODS: Wistar Albino rats were divided into 5 groups, each including 8 rats. The sham group underwent laparotomy only, while the Non-Epo-3 and Non-Epo-7 groups underwent common bile duct ligation and were sacrificed 3 and 7 days, respectively, after the operation. The Epo-3 and Epo-7 groups underwent common bile duct ligation and Epo treatment and were sacrificed 3 and 7 days, respectively, after the operation. Blood and tissue samples were collected from all groups for the determination of oxidative injury and hepatocellular damage. Serum total and direct bilirubin levels, alkaline phosphatase, reduced glutathione (GSH), nitric oxide (NO), malondialdehyde (MDA) and white blood cell counts were measured. RESULTS: Significantly higher NO and MDA levels were found in Non-Epo groups than Epo groups. Significantly lower GSH levels were found in the Non-Epo-7 group than the Epo-7 and sham groups. Hepatocellular damage was also found to be reduced in Epo groups. CONCLUSIONS: In the present model, while common bile duct ligation increased oxidative injury and hepatocellular damage, treatment with Epo attenuated oxidative injury and hepatocellular damage by decreasing NO and increasing GSH. PMID- 19556798 TI - Effect of tempol, a membrane-permeable radical scavenger, on mesenteric blood flow and organ injury in a murine cecal ligation and puncture model of septic shock. AB - BACKGROUND: Interventions that reduce the generation or the effects of reactive oxygen species exert beneficial effects in a variety of models of septic shock. We investigated the effect of tempol, a low-molecular-weight membrane-permeable radical scavenger, on mesenteric blood flow and organ injury in a murine cecal ligation and puncture (CLP) model of septic shock. MATERIALS AND METHODS: Forty four Swiss albino mice were anesthetized with chloral hydrate (400 mg/kg, i.p.) and subjected to CLP (except for the sham-operated animals). The animals were divided randomly into 4 groups: the 1st group was sham operated (sham-operated group, n = 10); the 2nd group underwent CLP and was injected with saline (CLP + saline group, n = 12); the 3rd group was sham operated and treated with tempol (10 mg/kg, i.p., sham-treated + tempol group, n = 10); the 4th group underwent CLP and was treated with tempol (10 mg/kg, i.p., CLP + tempol group, n = 12). Mesenteric arterial blood flow (MABF) was measured by Doppler ultrasound. Poly(adenosine 5'-diphosphate-ribose) polymerase (PARP) activity was examined in the liver, lung, and kidneys. RESULTS: In the CLP + saline group, the MABF was significantly lower than in the sham-operated group (p < 0.001). After tempol administration, MABF values significantly increased (p < 0.05). We observed significantly stronger PARP-positive staining in the lungs and kidney glomeruli in the CLP + saline group than in those of the sham-operated group (p(lung) = 0.0148, p(glomeruli) = 0.0025). A marked reduction in PARP activity was found in the lung and kidney glomeruli of the CLP + tempol group (p(lung) = 0.0026, p(glomeruli) = 0.0085). There was no significant effect of CLP on PARP activity in the liver and kidney tubuli (p(liver) > 0.05, p(tubuli) > 0.05). CONCLUSION: Tempol improved MABF in a CLP-induced septic shock model. Although tempol could not prevent the activation of PARP in the liver and kidney tubuli, it did attenuate PARP activation in the lung and kidney glomeruli. PMID- 19556800 TI - How to prevent lateral thermal damage to tissue using the harmonic scalpel: experimental study on pig small intestine and abdominal wall. AB - INTRODUCTION: When using a harmonic scalpel, the lower amount of energy that is transduced to the tissue reduces the chance of lateral thermal damage. METHODS: Pigs (weight: 40 kg) were used as the experimental model. After anesthesia, tissue was coagulated using different application regimens for each group. The width of tissue necrosis was measured from the point of incision by the harmonic scalpel. RESULTS: The pig abdominal tissues suffered mean thermal damage of 0.0825 (output power 3) and 0.2969 mm (output power 5) when used for 5 s; at 10 s these values were 0.3850 and 0.4793 mm, respectively. In a third experimental condition, with 10 s of application broken down into 2 parts of 5 s with a 5 second pause in-between, these values were 0.1876 and 0.2013 mm, respectively. The small intestine tissues suffered mean thermal damage of 0.1302 (output power 3) and 0.1771 mm (output power 5) at a duration of 5 s. After 10 s of application, these values changed to 0.2655 (output power 3) and 0.2983 mm (output power 5). In the third condition (activity for 5 s, pause for 5 s, activity for 5 s), they were 0.2011 and 0.2258 mm, respectively. CONCLUSION: Coagulation necrosis is bigger if the usage is continuous rather than if it is disconnected/reconnected. PMID- 19556802 TI - Cellular receptors for foot and mouth disease virus. AB - Foot-and-mouth disease virus (FMDV), the prototype member of the Aphthovirus genus, is a single-stranded, positive-sense RNA genome virus, which affects many domestic livestock cloven-hoofed animals, causing substantial lost of milk in dairy cattle, reduction in the growth rate of meat animals, among others. It has been shown that the virus can enter to the cells using different pathways; the main one binding integrins via the clathrin-mediated endocytosis pathway, trafficking throughout the acidified endocytic vesicles, where its capsid rapidly dissociates, resulting in the release of the RNA genome, and the second one using heparan sulfate in which FMDV enters to the cells using the caveola-mediated endocytosis pathway and that caveolae can associate and traffic with endosomes. Different integrins had been involved as FMDV receptors (alphavbeta1, alphavbeta3, alpha5beta1, alphavbeta6, alphavbeta8); this review will try to resume the basic information about FMDV receptors from the last years to the present and will resume the most important in vitro and in vivo studies to elucidate the role of this receptor on the infection. PMID- 19556801 TI - Interval of injections of intramuscular depot medroxyprogesterone acetate in the long-term treatment of endometriosis-associated pain: a randomized comparative trial. AB - AIM: To determine the optimal interval of injections of intramuscular depot medroxyprogesterone acetate 150 mg in the long-term treatment of endometriosis associated pain. METHOD: 112 patients with symptomatic endometriosis were randomized to receive either injections every month for 6 months, then every 3 months for a total of 15 months or injections every 3 months for 15 months. The primary outcome measure was patients' satisfaction. RESULT: At months 3, 6, 9, 12 and 15 of the treatment phase, there was no statistically significant difference of percentages of patients with satisfaction between the two regimens (85.7 vs. 76.8%, 76.8 vs. 73.2%, 66.1 vs. 58.9%, 60.7 vs. 55.4%, 60.7 vs. 55.4%, respectively). CONCLUSION: The optimal interval of injections of depot medroxyprogesterone acetate 150 mg is every 3 months. PMID- 19556803 TI - Evaluation of the hemodynamic status of focal hepatic lesions 20 mm or less in diameter by contrast-enhanced ultrasonography using Sonazoid. AB - OBJECTIVE: To clarify the validity of the evaluation of the hemodynamics of hepatic lesions < or =20 mm in diameter using contrast-enhanced ultrasonography (CE-US) with Sonazoid. METHODS: Sixty-two hepatic lesions in 55 patients with chronic hepatitis or cirrhosis due to hepatitis C virus were studied. We evaluated by quantitative analysis the time intensity curve (TIC) on hepatic lesion and parenchyma in the early vascular phase and Kupffer imaging in the post vascular phase. RESULTS: TIC patterns were classified into those with a maximum slope (Max slope) steeper in the hepatic lesion than in the parenchyma (Pattern I), those with a Max slope similar in the hepatic lesion and parenchyma (Pattern II), and those with a Max slope gentler in the hepatic lesion than in the parenchyma (Pattern III). The blood flow was considered to be higher, the blood flow velocity to be faster, and the contrast agent to reach the lesion more rapidly in Pattern I lesions than in the hepatic parenchyma. Pattern III lesions showed that the velocity of arterial blood influx was slow. CONCLUSION: Our study suggested the possibility that the TIC allows a detailed evaluation of the hemodynamics of hepatic lesions. PMID- 19556804 TI - Adaptation of skeletal muscle microvasculature to increased or decreased blood flow: role of shear stress, nitric oxide and vascular endothelial growth factor. AB - This review elucidates the roles of capillary haemodynamics, nitric oxide (NO) and vascular endothelial growth factor (VEGF) in the remodelling of skeletal muscle microcirculation in response to increased (electrical stimulation) or decreased (chronic ischaemia) blood flow. During early stages of stimulation induced angiogenesis, up-regulation of VEGF and its receptor VEGF receptor 2 is dependent on shear stress and NO release, whereas later, involvement of NO in the expanding capillary bed appears to be VEGF/VEGF receptor 2 independent. Arteriolar growth most likely relies on mechanical wall stresses while growth factor involvement is less clear. By contrast, in muscles with restricted blood flow, increased VEGF/VEGF receptor 2 expression after ischaemia onset is not associated with changes in shear stress or hypoxia, or capillary growth. After several weeks, VEGF protein levels are lower than normal while modest angiogenesis takes place, a temporal mismatch that limits the utility of using growth factor levels during ischaemia to assess angiogenic potential. Chronic stimulation of ischaemic muscles restores their depressed endothelial-dependent arteriolar dilatation, increases capillary shear stress and VEGF receptor 2 and promotes capillary growth. In patients with peripheral vascular disease, electrical stimulation of ischaemic calf muscles increases blood flow, capillary surface area and muscle performance, offering an alternative 'endogenous' treatment to gene or cell therapy. PMID- 19556805 TI - Predictors of higher blood pressure in a clinical setting in normotensive children: a prospective study. AB - This study was designed to assess the predictors of a higher blood pressure (BP) in a clinical setting in a prospective cohort of normotensive children recruited in an outpatient pediatric clinical setting in Southern Brazil. The study evaluated 443 children 3-12 years of age. If the mean value of BP was greater than the 95th percentile for age, sex and height in the medical setting, children were scheduled to other two evaluations at home. The systolic BP percentile in the clinical consultation was associated with BMI and birth weight. The diastolic BP in the clinical consultation was associated with birth weight, age, and BMI. Weight excess, low birth weight and younger age were associated with a higher BP in normotensive children seen in a clinical setting. To our knowledge, this is the first study to describe the association between lower birth weight and a greater BP response to medical consultation. Our finding suggests that the tendency to higher white-coat effect is determined, at least to some extent, in intrauterine life. PMID- 19556806 TI - Prevalence of primary focal or segmental dystonia in adults in the district of foggia, southern Italy: a service-based study. AB - BACKGROUND: Primary focal or segmental dystonia is a rare clinical condition including early-onset dystonia, which has the tendency to generalize, and late onset dystonia, which may be focal or segmental. The prevalence of late-onset dystonia ranges from 30 to 7,320 cases per million, but no data are available in Italy. METHODS: A service-based study was conducted in the period 1 January 2001 through 31 December 2002 in the administrative district of Foggia, southern Italy (population 541,653). Cases were traced through hospital discharge diagnosis, botulinum toxin services, day hospital access, ear, nose and throat, ophthalmology and orthopedic surgery specialists, and territorial outpatient services. Inclusion criteria were age 17 years or older, residency in the study area and a diagnosis of primary focal/segmental dystonia. RESULTS: A total of 69 patients were included, giving a crude prevalence of 127.4 per 1,000,000 (women: 146.4; men: 107.0; age 18-34 years: 39.2; 35-54 years: 98.7; 55-74 years: 273.6; 75+ years: 163.3). The standardized rate was 137.5 (95% confidence interval 107.0 174.6). Blepharospasm was the commonest clinical condition (prevalence 68.2), followed by cervical dystonia (prevalence 44.8). CONCLUSION: The prevalence of primary focal or segmental dystonia in Italy is in keeping with several other reports, but is lower than in studies performed in northern Europe, Minnesota, USA, and Japan. The difference in our results may be mostly explained by misdiagnosis, underascertainment of cases and a fairly limited observation period. PMID- 19556807 TI - Occurrence of depression in families with frontotemporal dementia: a family history study. AB - BACKGROUND: There are clinical similarities between frontotemporal dementia (FTD) and depression. The aim is to study co-aggregation of depression in families with FTD, indicating the existence of common aetiological factors. METHODS: The study included 74 index patients with FTD and their 540 first-degree relatives above the age of 15 years. Occurrence of depression was studied at 3 different levels. RESULTS: The incidence of depression in first-degree relatives of FTD patients was not higher than that of a general population. Occurrence of depression was not higher in families where parents had FTD compared to families with parents having no indications of FTD. Individuals with FTD had not suffered from depression to a greater extent than those without FTD. CONCLUSIONS: The hypothesis of a common aetiological factor of FTD and depression was not supported. PMID- 19556808 TI - Current oncological treatment of patients with pancreatic cancer in germany: results from a national survey on behalf of the Arbeitsgemeinschaft Internistische Onkologie and the Chirurgische Arbeitsgemeinschaft Onkologie of the Germany Cancer Society. AB - BACKGROUND: No data have previously been available regarding the current treatment of patients with pancreatic cancer (PC) in German hospitals and medical practices. METHODS: Between February 2007 and March 2008 we conducted a national survey [on behalf of the Arbeitsgemeinschaft Internistische Onkologie (AIO) and the Chirurgische Arbeitsgemeinschaft Onkologie (CAO)] regarding the current surgical and oncological treatment of PC in Germany. Standardized questionnaires were sent via mailing lists to members of the AIO and CAO (n = 1,130). The data were analyzed using SPSS software (version 16.0). Pre-defined subgroup analysis was performed by grouping the results of each question with regard to the professional site of the responding physician and to the number of patients treated in their institution by year. RESULTS: 181 (16%) of the oncological questionnaires were sent back. For 61% of the participating centers, a histological confirmation of PC diagnosis is obligatory. 21% of physicians offer neoadjuvant therapy to patients with potentially resectable PC. In the adjuvant treatment after curative-intent surgery, gemcitabine (Gem) is regarded as standard of care by 71% after R0 resection and 62% after R1 resection. For patients with locally advanced PC, 52% of the participating centers recommend systemic chemotherapy, 17% prefer combined primary chemoradiotherapy. Most centers (59%) base their decision of combination regimens for metastatic disease on the performance status of their patients. In patients with a good status, 28% apply single-agent Gem, 3% use Gem + capecitabine, 12% Gem + erlotinib, 16% Gem + oxaliplatin, and 8% Gem + cisplatin. Only 28% of the survey doctors offer second line treatment to the majority of their patients with advanced PC. CONCLUSION: Not every PC patient in Germany is treated according to the present S3 guidelines. Diagnosis and treatment of PC in Germany still need to be improved. PMID- 19556809 TI - Phase II study of biweekly docetaxel and S-1 combination therapy for advanced or recurrent gastric cancer. AB - OBJECTIVE: This phase II study evaluated the toxicity and efficacy of a novel dosing schedule of docetaxel and S-1 as treatment for advanced gastric cancer. METHODS: Patients with measurable advanced or recurrent gastric cancer and no prior exposure to the investigational drugs were treated with intravenous docetaxel 35 mg/m(2) on days 1 and 15, and oral S-1 80 mg/m(2)/day on days 1-14 every 4 weeks. The primary endpoint was objective response. RESULTS: Thirty-five eligible patients were enrolled and received a total of 151 cycles of treatment (median 3, range 1-19). One complete response and 13 partial responses were observed, with an overall response rate of 40% (95% CI: 24-56%). Median progression-free survival and median overall survival times were 4.5 and 14.2 months, respectively. The most common grade 3-4 toxicities were neutropenia (23%) and leukocytopenia (15%). CONCLUSION: Biweekly docetaxel combined with S-1 is active in advanced or recurrent gastric cancer, and can be administered with proper management of adverse events in an outpatient clinic. PMID- 19556810 TI - Podoplanin expression identified in stromal fibroblasts as a favorable prognostic marker in patients with colorectal carcinoma. AB - OBJECTIVE: The microenvironment of cancer plays a critical role in its progression. However, the molecular features of cancer-associated fibroblasts (CAFs) are less well understood than those of cancer cells. We investigated the clinicopathological significance of podoplanin expression in stromal fibroblasts in patients with colorectal cancer (CRC). METHODS: We selected podoplanin as an upregulated marker in CAF from a DNA microarray experiment. Consequently, podoplanin was identified as an upregulated gene. Immunohistochemical podoplanin expression was investigated at the National Cancer Center Hospital, Tokyo, Japan, in 120 patients with advanced CRC, and its clinicopathological significance was examined. The biological function of podoplanin expression was also assessed by a coculture invasion assay with CRC cell lines such as HCT116 and HCT15. RESULTS: Podoplanin expression was exclusively confined to stromal fibroblasts and absent in tumor cells. Podoplanin is absent in normal stroma except for lymphatic vessels. Staining was considered positive when over 30% of the cancer stroma was stained. Positive podoplanin expression was significantly correlated with a more distal tumor localization (p = 0.013) and a shallower depth of tumor invasion (p = 0.011). Univariate analysis revealed that negative podoplanin expression in stromal fibroblasts was significantly associated with reduced disease-specific survival (p = 0.0017) and disease-free survival (p < 0.0001). Multivariate analysis revealed that negative podoplanin expression (p = 0.016) and lymph node metastasis (p = 0.027) were significantly associated with disease-free survival. CRC cell invasion was augmented by co-culture with CAFs that were treated with siRNA for podoplanin. CONCLUSIONS: Our results suggest that a positive podoplanin expression in stromal fibroblasts could have a protective role against CRC cell invasion and is a significant indicator of a good prognosis in patients with advanced CRC, supported by biological analysis showing that podoplanin expression in CAFs is associated with decreased CRC cell invasion. PMID- 19556811 TI - Phase I study of prolonged-infusion gemcitabine combined with cyclophosphamide in patients with metastatic carcinoma of the breast: tolerability of an optimal dose schedule. AB - BACKGROUND: A phase I study was initiated to determine the maximum tolerated dose (MTD) of prolonged-infusion gemcitabine combined with cyclophosphamide in patients with metastatic breast carcinoma (MBC). METHODS: Patients with MBC were treated with gemcitabine infusion at 10 mg/m2/min and cyclophosphamide by intravenous piggyback injection, 4 h after initiation of the infusion. We treated 3-6 patients at a particular dose level until the MTD was determined. RESULTS: Overall, 44 patients received a total of 197 courses of therapy. Both drugs were given on days 1, 8 and 15 to 14 patients (68 courses). Delayed white blood cell recovery necessitated first protocol amendment to drop cyclophosphamide on days 8 and 15 in 9 patients (43 cycles). A second amendment was needed to drop gemcitabine on day 15 because of thrombocytopenia in 21 patients (86 courses). The dose-limiting toxicity was thrombocytopenia. The MTD of an optimal dose schedule was 800 mg/m2 gemcitabine infused at a rate of 10 mg/m2/min on days 1 and 8, and 400 mg/m2 cyclophosphamide, by intravenous piggyback injection, on day 1, 4 h after initiation of the gemcitabine infusion. CONCLUSIONS: The MTD can be given safely every 4 weeks to patients with MBC. Phase II studies are warranted to evaluate the clinical activity of this therapy. PMID- 19556812 TI - S-1 monotherapy in patients with advanced biliary tract cancer. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of single-agent S-1 in patients with advanced biliary tract cancer. METHODS: S-1 was administered orally at a dose of 80 mg/m2 for 28 days, followed by 14 days of rest (1 cycle); treatment was repeated until disease progression, unacceptable toxicity, or patient refusal. RESULTS: Forty-five patients were enrolled between January 2005 and June 2008. Among these, 29 patients received S-1 as first-line chemotherapy and 16 patients received S-1 as second-line chemotherapy. The response rates for first- and second-line chemotherapy were 17.2 and 18.8%, respectively. The median times to progression for the first- and second-line chemotherapy groups were 4.2 and 5.5 months (p = 0.91), respectively. The median overall survival and 1-year survival rate for each group were 8.7 and 8.0 months and 42.2 and 38.2%, respectively (p = 0.62). Only the first-line chemotherapy group experienced grade 3/4 toxicities, including leukopenia (6.9%), neutropenia (10.3%), anemia (6.9%), thrombocytopenia (10.3%) and total bilirubin elevation (3.4%). CONCLUSION: S-1 monotherapy is a feasible and moderately efficacious treatment for advanced biliary tract cancer, as a first- or second-line chemotherapy regimen. PMID- 19556813 TI - Upping recruitment in clinical trials: are the costs worth it? PMID- 19556814 TI - Presentation, treatment, and analysis of prognostic factors of terminally ill patients with gastrointestinal tumors. AB - BACKGROUND: Few data have been published about terminally ill patients with gastrointestinal tumors treated in palliative care units. PATIENTS AND METHODS: We analyzed the data of 737 admissions of 435 patients that were treated in a palliative care unit, and tried to identify prognostic factors for survival. RESULTS: Most frequent diagnoses at admission were colorectal, gastric, esophageal, and pancreatic cancer. Major clinical symptoms were pain (66.9%), anorexia (60.8%), weight loss (39.2%), and nausea/vomiting (36.6%). In 71.6% of the patients, morphine derivatives were administered. In 33.0% of cases, red blood cell transfusions were applied, parenteral nutrition was given in 31.3%. Median survival, calculated from the day of first hospitalization, was 35 days. On univariate analysis, several clinical and laboratory parameters were identified as prognostically important factors. In the multivariate Cox regression analysis, 5 parameters were significant: ascites and anorexia, elevated leukocyte count and lactate dehydrogenase activity, as well as decreased albumine levels. Using these parameters, patients were divided into 3 risk groups: low-risk (presence of 0-1 factors), intermediate risk (2-3 factors), and poor-risk patients (4-5 factors). Median survival for poor-risk patients was 18 days, intermediate- and low-risk patients survived 43 and 136 days, respectively (p < 0.0001). CONCLUSION: In multivariate analysis, 5 prognostic factors were identified, and 3 patient groups were defined. After multicenter validation, these factors may help to guide treatment decisions in terminally ill patients with gastrointestinal tumors. PMID- 19556815 TI - Pro-brain natriuretic peptide is a sensitive marker for detecting cardiac metastases in patients with non-small cell lung cancer. AB - BACKGROUND: B-type natriuretic peptide (BNP) and N-terminal-pro-BNP (NT-pro-BNP) are important diagnostic tools for patients with suspected cardiac disorders. The aim of this study was to evaluate the predictive value of plasma NT-pro-BNP in identifying cardiac metastases in patients with non-small cell lung cancer (NSCLC) and dyspnoea. PATIENTS AND METHODS: A total of 120 patients, median age 62 years (range 46-83), with NSCLC and dyspnoea were studied. Patients with heart failure or documented coronary artery disease were excluded. Echocardiographic imaging was used to detect cardiac metastases and estimate global left ventricular function. Ejection fraction and E/A ratio from transmitral inflow pattern were calculated. Plasma NT-pro-BNP was also measured. 72 patients (72/120, 60%) with cardiac metastases were identified. RESULTS: NT-pro-BNP was significantly higher in patients with metastases (1347.5 +/- 1004.30 pg/ml vs. 159.02 +/- 93.29 pg/ml; p = 0.001). No differences between groups, regarding s creatinine (p = 0.45), haemoglobin (p = 0.71), left ventricular hypertrophy (p = 0.91), and diastolic dysfunction (p = 0.79), were observed. CONCLUSION: Plasma NT pro-BNP is remarkably elevated in patients with NSCLC and myocardial/pericardial infiltrations and may be used as a sensitive marker for detecting cardiac metastases in these patients. PMID- 19556816 TI - Hematoma after vacuum-assisted breast biopsy: are interleukins predictors? AB - BACKGROUND: Hematoma is the main complication of vacuum-assisted breast biopsy (VABB). This study aims to evaluate the associations between interleukin (IL) 1alpha, IL-1beta and IL-6 and hematoma progression. METHODS: This study included 36 women who underwent VABB (11G). After VABB, mammograms were obtained from these patients and the maximum diameter of the hematomas was measured. The hematoma progression / occurrence of organized hematomas was followed up for the subsequent 30 days. Venous samples were collected peripherally at 3 time points: prior, at the end, and 1 h after the end of the VABB procedure. Enzyme-linked immunosorbent assays were used for the determination of serum IL-1alpha, IL-1beta and IL-6 levels. RESULTS: 2/36 hematomas were eventually organized within the follow-up period. In these cases, IL-6 had been significantly higher 1 h after the end of VABB (5.70 +/- 0.18 vs. 1.73 +/- 1.01 pg/ml; p = 0.019, Mann-Whitney Wilcoxon test for independent samples). No statistically significant associations existed concerning IL-1alpha and IL-1beta. The association between the size of a hematoma on the mammogram and the subsequent organization did not reach statistical significance. CONCLUSIONS: Elevated IL-6 at 1 h after the end of VABB might point to subsequent organization of the hematoma and the need for appropriate action. PMID- 19556817 TI - Cisplatin as single-agent chemotherapy in patients with liver dysfunction due to metastases. AB - BACKGROUND: Chemotherapeutic options are often limited for patients with hepatic dysfunction induced by advanced metastases. The toxicity and efficacy of cytotoxic drugs are often unpredictable due to altered drug activation or inactivation and excretion. Therefore, numerous chemotherapeutic agents should not be administered to patients with liver failure. PATIENTS AND METHODS: We retrospectively analysed 14 patients with solid tumours presenting with liver dysfunction induced by metastasis who were treated with cisplatin single-agent chemotherapy. RESULTS: Tolerance of therapy was acceptable with 1 grade I and 1 grade II renal toxicity. 6 patients experienced grade III haematological toxicity. Partial remission of the disease was observed in 6 cases, and 6 patients could receive combination chemotherapy after improvement of liver function. The median overall survival of the patient cohort was 4.8 months. CONCLUSION: Cisplatin monotherapy may thus be considered as a treatment option for patients with liver dysfunction induced by different solid tumours. PMID- 19556818 TI - Coexistence of copy number increases of c-Myc, ZNF217, CCND1, ErbB1 and ErbB2 in ovarian cancers. AB - BACKGROUND: We selected 5 oncogenes with well-established roles in carcinogenesis -- CCND1, ErbB1, ErbB2, c-myc and ZNF217 -- to investigate the coexistence of their copy imbalances in relation to the clinico-pathological characteristics of ovarian tumors. MATERIALS AND METHODS: Fluorescence in situ hybridization for the 5 genes was applied to a preexisting tissue microarray. 38 ovarian tumors were successfully analyzed for copy number changes of the 5 genes. RESULTS: At least one of these oncogenes was gained/amplified in 27 out of 38 tumors (71.1%). We report the highest frequency of c-myc genetic gain/amplification since it affected 42.1% of the ovarian tumors. We observed sequential involvement of copy number alterations of the other genes in the presence of c-myc disruption. The incidence of copy number changes of the 5 oncogenes -- both single and combinatorial -- was higher in high-grade tumors. All double aberrations in the serous group comprised c-myc and ZNF217copy number increases. CONCLUSIONS: Our results revealed a combination between copy number increases of c-myc and ZNF217, associated with serous histology. The data from this combined analysis of the 5 oncogenes could be used as a basis in considering the combined approach in molecular-based therapy of ovarian cancer. PMID- 19556819 TI - Making or losing money with participation in clinical trials: a decision analysis. AB - BACKGROUND: There is increasing demand and quality-driven pressure from professional organizations for physicians and health care providers to increase participation in clinical studies. But this can have a severe financial impact on the institution, so costs should be identified and calculated in advance. METHOD: In a diagram, the decision-making process to participate in clinical trials based on economic and budget impact is reviewed and analyzed in detail. RESULTS: This flow chart describes how cost-effective participation in clinical trials is determined. Since its implementation, all trials in our institution have been cost covering. CONCLUSIONS: All service and care required within the studies must be distinguished as either medically necessary or study related. Costs for the first category have to be covered by the health care system, but in case of the second category by the study sponsor. The institution's own costs for study related services should be known and deducted from the study income to determine the actual study gains. Subsidizing studies from tight clinic budgets is difficult in times of rationed medicine and should be avoided. Non-cost-covering clinical studies should be re-negotiated with the sponsor until cost effectiveness is reached. Otherwise, a rejection of study participation for financial reasons should be seriously considered. The design of cost-covering clinical trials supports better recruitment for studies. PMID- 19556820 TI - Non-gestational uterine choriocarcinoma in a postmenopausal woman. AB - BACKGROUND: Gestational trophoblastic disease occurs rarely in postmenopausal women. CASE REPORT: We report on a 65-year-old woman with uterine choriocarcinoma developing 16 years after menopause and 25 years after her last pregnancy. She was found to have a uterine tumor on laparotomy after presenting with uterine bleeding and abdominal pain. Histopathological examination demonstrated malignant syncytiotrophoblastic and cytotrophoblastic cells with extensive necrosis and hemorrhage, consistent with pure choriocarcinoma. Chemotherapy consisting of etoposide, methotrexate, actinomycin D, cyclophosphamide, and vincristine (EMA CO) was started. The treatment was changed to methotrexate and folinic acid because of severe hypersensitivity reaction after etoposide infusion. After 4 cycles, the serum beta-human chorionic gonadotropin (beta-hCG) level had decreased to normal. The patient remains disease free 20 months after the treatment. CONCLUSIONS: This case further illustrates that choriocarcinoma may be seen in older women after a long menopausal period. Accurate diagnosis and treatment are essential, because the tumor is very chemosensitive and curable even in advanced stages. PMID- 19556821 TI - Hemicentral retinal artery occlusion in a breast cancer patient using anastrozole. AB - BACKGROUND: Anastrozole, an aromatase inhibitor, is commonly used in the adjuvant treatment of breast cancer. Anastrozole treatment is associated with a risk of thromboembolic events and retinal vascular side effects. Herein, we present a case of hemi-central retinal artery occlusion diagnosed in a breast cancer patient using anastrozole. CASE REPORT: A 53-year-old woman with a hypertensive and diabetic background was admitted to our hospital with breast cancer. Anastrozole treatment was started after surgery, adjuvant chemotherapy, and radiotherapy. Sudden painless loss of vision in the patient's right eye occurred within 13 months of Anastrozole treatment. A fluorescein angiogram revealed hemi central retinal artery occlusion. CONCLUSION: To the best knowledge of the authors, this is the first report of hemi-central retinal artery occlusion in an anastrozole user. PMID- 19556823 TI - Small cell lung cancer presenting as ectopic ACTH syndrome with hypothyroidism and hypogonadism. AB - BACKGROUND: Small-cell lung cancer accounts for 15-20% of all lung cancers, and it is the cell type most commonly associated with paraneoplastic syndrome. Small cell lung cancer presenting as ectopic adrenocorticotropic hormone (ACTH) syndrome associated with hypothyroidism and hypogonadotropic hypogonadism is clinically very rare. CASE REPORT: A 43-year-old man who presented with bilateral lower-extremity edema and hypokalemia had a mass lesion in his left hilum base visible on chest radiograph. Biopsy identified the mass as small-cell lung cancer with focal ACTH staining. The endocrine tests disclosed hypercortisolism, hypogonadism and hypothyroidism. RESULTS: Partial remission as evidenced by regression of the tumor mass and return to normal serum cortisol and ACTH levels occurred after the first course of combination chemotherapy using cisplatin and etoposide. An unexpected left-sided spontaneous pneumothorax developed after the first course of chemotherapy and was treated with thoracostomy and a chest tube. The patient developed persistent air leakage and chronic empyema. The patient received surgery of the Eloesser flap and reconstruction with the latissimus dorsalis flap. The treatment of the complicated problems was successful. CONCLUSION: Combination chemotherapy may prove effective in the treatment of small-cell lung cancer with ectopic ACTH syndrome, hypothyroidism and hypogonadism. PMID- 19556822 TI - Metastasis of lobular breast carcinoma to the uterus in a patient under anastrozole therapy. AB - BACKGROUND: Metastasis of extragenital neoplasms to the uterus are rarely encountered, and usually occur as a manifestation of advanced disease. Lobular carcinoma is the most common type of breast cancer that metastasizes to the uterus. CASE REPORT: We report on a 56-year-old woman who 3 years previously was diagnosed with invasive lobular carcinoma of the breast and was treated with surgery followed by chemotherapy and radiotherapy. While the patient was on adjuvant anastrozole therapy for 2 years, she complained of vaginal bleeding. Because of endometrial thickening and a uterine leiomyoma detected during abdominal ultrasonographic ex-amination, a total hysterectomy with bilateral salpingo-oophorectomy was performed. Histopathologic examination of the specimens revealed carcinoma infiltration of the myometrium, endometrium, cervix, uterine tube, and left ovary. Immunohistochemical staining of tumoral cells with pancytokeratin and gross cystic disease fluid protein (GCDFP-15) proved the diagnosis of metastatic lobular breast carcinoma to the uterus. CONCLUSION: To our knowledge, this is the second case of lobular breast carcinoma metastasized to the uterus under anastrozole therapy. In women with lobular breast cancer under adjuvant anastrozole therapy, who present with vaginal bleeding, uterine metastasis of lobular carcinoma should be considered in the differential diagnosis. PMID- 19556824 TI - Bevacizumab plus oxaliplatin-based regimens for the treatment of colorectal cancer. AB - The anti-angiogenic, monoclonal antibody bevacizumab, in combination with chemotherapy, is proven to prolong survival in metastatic colorectal cancer. Recent data demonstrate that combining bevacizumab with widely used oxaliplatin containing chemotherapy regimens is a promising strategy for further improving patient outcomes. PMID- 19556825 TI - Lymphomatoid granulomatosis in a patient with rheumatoid arthritis receiving methotrexate: successful treatment with the anti-CD20 antibody mabthera. PMID- 19556826 TI - Intraocular pressure-lowering activity of topical application of Aegle marmelos fruit extract in experimental animal models. AB - AIM: The present study was designed to evaluate the intraocular pressure (IOP) lowering activity of topical application of the aqueous extract of Aegle marmelos fruit in experimental animal models. MATERIALS AND METHODS: New Zealand white rabbits with normal and experimentally elevated IOP using water loading and steroid-induced models were included in this study. The IOP-lowering effect of A. marmelos fruit extract in rabbits with experimentally elevated IOP was also compared with that of timolol 0.25%. RESULTS: In rabbits with normal IOP, the A. marmelos fruit extract at a concentration of 1% showed the maximum IOP-lowering effect with 22.81% reduction from baseline IOP. The maximum IOP reduction achieved in water loading and steroid-induced models with the same concentration of A. marmelos was 27.57 and 28.41% from baseline, respectively. The efficacy was comparable to that of timolol after 45 min of water loading in the water loading model, and during the first 2 h of treatment in the steroid-induced model. CONCLUSION: A. marmelos fruit extract showed significant IOP-lowering activity in experimental animal models. PMID- 19556828 TI - Microscopy. AB - Microscopy employs the use of microscopes, which are instruments designed to produce magnified visual or photographic images of objects too small to be seen with the naked eye. The goal of the microscope is to accomplish three tasks, namely produce a magnified image of the specimen, separate the details in the image, and render the details visible to the human eye or camera. The development of microscopy revolutionized biology and, to this day, remains an essential tool in science. PMID- 19556827 TI - Association of TNFA -308 G/A and TNFRI +36 A/G gene polymorphisms with glaucoma. AB - OBJECTIVE: TNF-alpha is one of the most important factors recognized in the pathogenesis of open-angle glaucoma. Therefore, the association of single nucleotide polymorphisms in the TNFRI gene and the promoter region of the TNFA gene with glaucoma susceptibility was investigated in the present study. METHOD: In total, 223 patients with glaucoma and 202 unrelated controls were genotyped by allele-specific oligonucleotide PCR and PCR- restriction fragment length polymorphism to determine TNFA -308 G/A and TNFRI +36 A/G polymorphisms, respectively. RESULTS: In contrast to TNFRI polymorphisms, the genotypes of TNFA 308 G/A polymorphisms showed a significant difference between patients and controls (p = 0.0025). Of interest, the distribution of TNFA genotypes was significantly different between patients with primary open-angle glaucoma (p = 0.001) or pseudoexfoliative glaucoma (p = 0.001) and controls, while no difference was found when chronic angle-closure glaucoma patients were compared to controls (p = 0.72). CONCLUSION: In line with studies showing the role of TNF alpha in open-angle glaucoma, the results of the present study showed that inheritance of the high producer TNFA -308 A allele might be a susceptibility factor for the development of open-angle glaucoma. PMID- 19556829 TI - Genistein in the presence of 17beta-estradiol inhibits proliferation of ERbeta breast cancer cells. AB - BACKGROUND/AIM: Genistein, a soy component, has been shown to have a biphasic proliferative effect in breast cancer cells, inhibiting in vitro cell proliferation at high concentrations (>10 micromol/l), while stimulating cell proliferation at lower concentrations (<10 micromol/l). However, epidemiological studies have shown an inverse correlation between the intake of genistein and the incidence of breast cancer. One of the possible reasons for this discrepancy could be the differing status of the estrogen receptor (ERalpha and/or ERbeta). Genistein selectively binds to ERbeta with strong affinity and thereby could be a potential chemotherapeutic agent against breast cancer of the ERalpha-negative and ERbeta-positive type. Therefore, the objective of the present study was to determine whether the proliferative effects of genistein were caused by its activity as a selective ERbeta agonist or merely as an antiestrogen. METHOD: This study was carried out in MDA-MB-231 (ERbeta) and T47D (ERalpha and ERbeta) human breast cancer cells. Cell proliferation was determined by the MTT (3-[4,5 dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide) assay. The cells were grown in estrogen-starved media and exposed to genistein at different concentrations for 72 h, either in the presence or absence of 17beta-estradiol. RESULTS: A significant decrease in cell proliferation was seen in MDA-MB-231 cells at low concentrations of genistein in the presence of 17beta-estradiol, as compared to genistein alone. In T47D cells, which are known to have a predominance of ERalpha over ERbeta, genistein showed a biphasic cell proliferative response both in the presence and absence of 17beta-estradiol. CONCLUSIONS: Our results suggest that in cells with a predominance of ERalpha, genistein acts as an agonist to ERalpha, and in cells with ERbeta alone, genistein most likely acts as an antiestrogen. Our results also suggest that genistein could be useful as a chemotherapeutic agent in premenopausal women with breast cancer of the ERalpha-negative and ERbeta-positive type. PMID- 19556830 TI - CT/MR image fusion in the postoperative assessment of electrodes implanted for deep brain stimulation. AB - BACKGROUND/AIMS: Stereotactic postoperative imaging is essential for verification of the position of electrodes implanted for deep brain stimulation (DBS). MRI offers superior visualisation of the DBS targets relative to CT, but previous adverse incidents have heightened concerns about risks of postoperative MRI. Preoperative MRI fused with postoperative CT offers an alternative method for evaluating electrode position, but before this method can be clinically applied, the image registration accuracy must be established. The purpose of this study was to quantitatively assess the accuracy of three different image registration and fusion methods. METHODS: Preoperative stereotactic MRI and postoperative stereotactic CT were acquired from 20 patients under- going DBS surgery (35 electrodes in total). The postoperative CT was registered and fused with the preoperative MRI, using three different registration algorithms. The position of each electrode tip was determined in stereotactic coordinates both in the (unfused) postoperative CT and the fused CT/MRI. The difference in tip position between the CT and fused CT/MRI was used to evaluate the registration accuracy. RESULTS: The mean error along the lateral, anteroposterior, and vertical axes was 0.5, 0.5, and 1 mm, respectively. CONCLUSIONS: CT/MRI fusion provides a safe, practical technique for postoperative identification of DBS electrodes. PMID- 19556831 TI - Stereotactic atlas-based depth electrode localization in the human amygdala. AB - BACKGROUND: The efficacy of stereotactic neurosurgery procedures is critically dependent on the accuracy of the device placement procedure. The first step in this process involves correctly identifying the target location in three dimensional brain space. In some clinical applications, this targeting process cannot be accomplished using MRI images of gross anatomical structures alone. The amygdala complex is a case in point, in that it consists of multiple histologically defined subnuclei with different functional characteristics. METHODS: In this report, we describe an elastic atlas brain-morphing method that projects amygdala subnuclear anatomical information onto the MRI volumes of individual subjects. RESULTS: The accuracy of this method was tested in 5 representative subjects using quantitative image-matching analytical techniques. The results demonstrate a high degree of intersubject variability in medial temporal lobe anatomy, and markedly superior anatomical matching performance by the elastic morphing method compared to Affine transformation. CONCLUSION: Nonlinear elastic morphing technique provides superior performance on fitting atlas templates to individual brain. The strengths and limitations of this and other atlas morphing methods are discussed in the context of emerging functional neurosurgery applications. PMID- 19556832 TI - Automated 3-dimensional brain atlas fitting to microelectrode recordings from deep brain stimulation surgeries. AB - OBJECTIVE: Deep brain stimulation (DBS) surgeries commonly rely on brain atlases and microelectrode recordings (MER) to help identify the target location for electrode implantation. We present an automated method for optimally fitting a 3 dimensional brain atlas to intraoperative MER and predicting a target DBS electrode location in stereotactic coordinates for the patient. METHODS: We retrospectively fit a 3-dimensional brain atlas to MER points from 10 DBS surgeries targeting the subthalamic nucleus (STN). We used a constrained optimization algorithm to maximize the MER points correctly fitted (i.e., contained) within the appropriate atlas nuclei. We compared our optimization approach to conventional anterior commissure-posterior commissure (AC/PC) scaling, and to manual fits performed by four experts. A theoretical DBS electrode target location in the dorsal STN was customized to each patient as part of the fitting process and compared to the location of the clinically defined therapeutic stimulation contact. RESULTS: The human expert and computer optimization fits achieved significantly better fits than the AC/PC scaling (80, 81, and 41% of correctly fitted MER, respectively). However, the optimization fits were performed in less time than the expert fits and converged to a single solution for each patient, eliminating interexpert variance. CONCLUSIONS AND SIGNIFICANCE: DBS therapeutic outcomes are directly related to electrode implantation accuracy. Our automated fitting techniques may aid in the surgical decision-making process by optimally integrating brain atlas and intraoperative neurophysiological data to provide a visual guide for target identification. PMID- 19556833 TI - Prognostic factors of subthalamic stimulation in Parkinson's disease: a comparative study between short- and long-term effects. AB - BACKGROUND/AIMS: Bilateral subthalamic nucleus deep brain stimulation (STN-DBS) has been shown to have long-term benefits in Parkinson's disease (PD). Through analyzing different variables, this study identified prognostic factors for the short- and long-term effects of STN-DBS. METHODS: Thirty-six PD patients underwent bilateral STN-DBS. Clinical evaluations were performed 1 month before and 3 months after surgery, with additional follow-up examinations for a mean of 31.3 months. RESULTS: There was a trend for long-term STN-DBS-induced improvements in the Unified Parkinson's Disease Rating Scale (UPDRS) part II and part III measures to be greater in younger patients. Preoperative levodopa responsiveness only led to consistent UPDRS part III improvement from STN-DBS at 3 months, and this predictive value did not exist in the long term. The preoperative levodopa response of tremor and axial symptoms in motor disability predicted long-term DBS effect only. Preoperative cognitive function positively correlated with postoperative improvement from DBS in UPDRS part III during long term follow-up only. CONCLUSIONS: The prognostic factors for STN-DBS benefit were different for short- and long-term follow-ups. Good prognostic factors for long term STN-DBS for PD patients were good cognitive function and tremor dominance. Poor prognostic factors were related to older age and non-dopaminergic-responsive axial disability. PMID- 19556834 TI - Dorsal root entry zone lesions for phantom limb pain with brachial plexus avulsion: a study of pain and phantom limb sensation. AB - BACKGROUND: Lesions in the dorsal root entry zone (DREZ) have been shown to be significantly effective in relieving the pain of brachial plexus avulsion (BPA), but they have a limited effect on phantom limb pain (PLP). There is still the question remaining of whether DREZ lesions are effective in treating PLP in cases of BPA. METHODS: Our study includes 14 post-traumatic patients with BPA and upper limb amputation. All developed PLP and underwent DREZ lesions. After the surgery, patients were asked to estimate the global percent of pain relief (0-100%). The phantom limb sensation (PLS) was also inquired after. RESULTS: Overall, 9 (64.3%) of 14 patients had satisfactory pain relief; the mean follow-up was 15.2 +/- 6.6 months. Among the 9 patients with satisfactory pain relief, PLS had altered in 6 (66.7%), while, in the other 5 patients with poor pain relief, none had experienced alterations in PLS. CONCLUSION: DREZ lesions are effective in the treatment of PLP with BPA. Alteration in PLS after the surgery may be a predictive factor for good pain relief. The good response of PLP patients with BPA to DREZ lesions suggests that an evaluation of the cervical dorsal roots should be conducted in patients with post-traumatic PLP. PMID- 19556836 TI - Deep brain stimulation for schizophrenia. PMID- 19556838 TI - [Pancreatic sphincter of oddi dysfunction]. PMID- 19556837 TI - DBS for neurobehavioral disorders. PMID- 19556839 TI - [Natural course and treatment strategy of gallbladder polyp]. AB - The polypoid lesions of gallbladder have explosively increased with enhanced feasibility of transabdominal ultrasonography. Most of small polyps less than 10 mm are benign and remain static for a long period. In small polyps, three to six month intervaled ultrasonography is warranted in the initial follow-up, but the duration of follow-up period is not clarified. The polypoid lesions larger than 10 mm show a quite different feature. They showed a remarkable risk of malignancy (34-88%) and should be treated by surgery. Furthermore, age more than 50 years and combined gallstone are important factors predicting malignancy in polypoid lesions of gallbladder. In addition, other factors including solitary polyp and the presence of symptoms are considered as risk factors. Laparoscopic cholecystectomy is a golden standard therapy for these polyps unless the suspicion of malignancy is high. The gallbladder polyps remain a problem of concern to both doctors and patient with the worry of malignancy. Thus, the comprehensive understanding of natural coruse of gallbladder polyp and risk factors of malignancy should be kept in mind. PMID- 19556835 TI - The hippocampus and nucleus accumbens as potential therapeutic targets for neurosurgical intervention in schizophrenia. AB - Schizophrenia is a chronic and disabling psychiatric illness that is often refractory to treatment. Psychotic symptoms (e.g. hallucinations and delusions) in schizophrenia are reliably correlated with excess dopamine levels in the striatum, and have more recently been related to excess metabolic activity in the hippocampus. Multiple lines of evidence suggest that aberrantly high hippocampal activity may, via hippocampal connections with the limbic basal ganglia, drive excessive dopamine release into the striatum. In the present paper, we hypothesize that inhibition or stabilization of neural activity with high frequency electrical stimulation of the hippocampus or nucleus accumbens, through different mechanisms, would treat the positive symptoms of schizophrenia. Thus, we suggest a direction for further experimentation aimed at developing neurosurgical therapeutic approaches for this devastating disease. PMID- 19556840 TI - [The prevalence of Helicobacter pylori infection in Korean patients with inflammatory bowel disease, a multicenter study]. AB - BACKGROUND/AIMS: The prevalence of Helicobacter pylori (H. pylori) infection has been reported to be lower in individuals with inflammatory bowel disease (IBD) in some Western countries. We investigated H. pylori infection in Korean patients with IBD and any possible associations of H. pylori infection with drug therapy for IBD and the phenotype of Crohns disease (CD). METHODS: We studied 316 unselected patients with IBD, including 169 ulcerative colitis (UC) patients and 147 with CD, and the control group consisted of 316 age- and gender-matched healthy people who received a comprehensive medical examination for a regular checkup purpose. Infection rates of H. pylori as detected by the urea breath test were compared between the IBD patients and the controls. RESULTS: A statistically significant difference in H. pylori infection rate was noticed between the IBD patients (25.3%) and the controls (52.5%; p<0.001), and between UC (32.0%) and CD patients (17.7%; p=0.04). Among the IBD patients, the age group of <60 and individuals with a history of taking metronidazole (13.0%; p=0.038) or ciprofloxacin (6.7%; p=0.001) were found to have a meaningfully lower infection rate, but those who did not take antibiotics still showed H. pylori infection rate significantly lower than the controls (CD 22.0% vs. UC 33.8% vs. Control 52.5%, p<0.001). With an exception of age, phenotypic characteristics showed no significant relations with H. pylori infection rate in CD patients. CONCLUSIONS: Korean patients with IBD, particularly CD, were found to have a significantly lower H. pylori infection rate than the controls. This association was more evident in those <60 years old, which suggested that H. pylori infection might be deemed to lower possible risks of IBD in younger adults. PMID- 19556841 TI - [Association of lower urinary tract symptoms with irritable bowel syndrome in adult men - an internet-based survey]. AB - BACKGROUND/AIMS: Patients with irritable bowel syndrome (IBS) are more likely to experience certain urinary symptoms. The aims of this study were to investigate the association between lower urinary tract symptoms (LUTS) and IBS, and to determine their impact on the quality of life. METHODS: E-mails were sent to 23,594 men who were registered at an internet survey company. Subjects were requested to fill out the questionnaires regarding IBS and LUTS assessed by the International Prostate Symptom Score (IPSS). RESULTS: Among 601 subjects (mean age, 35.5+/-8.4) included in the final analysis, 118 (19.6%) fulfilled the Rome II criteria for the diagnosis of IBS. The total mean IPSS of IBS subjects was 9.6, which was significantly higher than the 7.0 of non-IBS subjects (p<0.01). When IPSS was subcategorized into mild, moderate, and severe symptom categories, the proportions with the moderate and severe symptoms among IBS subjects were 33.9% and 13.6% respectively, which were significantly higher than those of non IBS subjects; (26.9 and 5.2%) (p<0.01). In multivariate analysis, statistically significant association was found between IBS and moderate to severe LUTS (OR, 1.91; 95% CI, 1.27-2.88). IBS subjects also showed a poorer quality of life score than non-IBS subjects (2.24 vs. 1.65, p< 0.001). CONCLUSIONS: LUTS in adult men with IBS are more severe and have a more negative impact on the quality of life than in non-IBS subjects. PMID- 19556842 TI - [A study on viral hepatitis markers and abnormal liver function test in adults living in northwest area of Chungnam]. AB - BACKGROUND/AIMS: We studied the prevalence of chronic hepatitis B, C and abnormality on liver function among the population in northwest area of Chungnam. METHODS: We have reviewed 40,112 adults who had received medical examination at health promotion center in Dankook university hospital. We studied them retrospectively about HBsAg, HBsAb, anti-HCV, and liver function test (LFT). RESULTS: Among the study subjects, 22,936 men and 17,176 women were involved. The overall seroprevalence of HBsAg was 4.2%. The prevalence in men (4.5%) was higher than that of women (3.7%) (p<0.001). The seroprevalence of HBsAg in their age was 5.1% in the 5th decade, 4.2% in the 2nd decade, 4.1% in the 4th decade, and 4.1% in the 6th decade. The overall seroprevalence of HBsAb was 65.1%. The overall seroprevalence of anti-HCV was 0.7%. After we reexamined them with HCV RNA or RIBA (Recombinant Immunoblot Assay), the prevalence of chronic hepatitis C was 0.09%. The LFT abnormality in total subjects was 11.4%. The LFT abnormality of chronic hepatitis B and C subjects was 21.72% and 63.2%. CONCLUSIONS: The prevalence of chronic hepatitis B and C was lower than that of previous studies. The prevalence of chronic hepatitis B in the 2nd decade was still high. PMID- 19556844 TI - [A case of left paraduodenal hernia combined with acute small bowel obstruction]. AB - Paraduodenal hernias are rare congenital malrotational anomalies of midgut that arise in the potential spaces and folds of the posterior parietal peritoneum adjacent to the ligament of Treitz and can lead to intestinal obstruction. Also, they have shown several presentation patterns, such as asymptomatic, chronic intermittent abdominal pain, and acute severe abdominal pain. If symptomatic hernias with strangulation are untreated, according to the previous reports, they lead to overall mortality exceeding 50%. We report a case of the left paraduodenal hernia combined with small bowel obstruction from the patient who had no history of surgery before and developed abdominal pain after drinking of alcohol heavily. Abdominal CT scan showed sac-like mass of clustered in the left upper quadrant. The patient underwent the surgery of the bowel reduction and adhesiolysis and got uneventful recovery. PMID- 19556843 TI - [Analysis of gene expression in primary hepatocellular carcinoma using differentially displayed reverse transcriptase polymerase chain reaction]. AB - BACKGROUND/AIMS: The investigation of a specific tumor marker for hepatocellular carcinoma (HCC) is needed to examine the carcinogenesis and to select the patients for treatment options. The aim of this study was to find the genes related to HCC. We also examined the expression level of these genes in cancer cell lines and tissue specimens. METHODS: Three pairs of HCC tissue and non neoplastic hepatic tissue around the HCC were collected from three patients who underwent resection for HCC. Differential display reverse transcriptase-PCR (DD RT-PCR) using GeneFishingTM PCR was used to detect the differences in the gene expression between in HCC tissue and non-neoplatic tissue. Up- or down-regulated genes in HCC tissue were identified through BLAST searches after cloning and sequencing assays. Real-time RT-PCR assay was employed to detect the expression rate in 11 HCC tissues and human cancer cell lines. RESULTS: Differentially expressed 21 genes were identified, and they were classified as genes involved in protein metabolism, ubiquitin-dependent protein catabolism, carbohydrate metabolism, lipid metabolism, DNA repair, and inflammatory response. CONCLUSIONS: We identified differentially expressed genes in HCC, and these genes may play an important role in the study of hepatocarcinogenesis, development of biomarker, and target therapy for HCC. PMID- 19556845 TI - [Metachronous four primary malignancies in gastro-intestinal tract]. AB - Multiple primary malignancy was reported firstly by Billroth in 1889. Recently, multiple primary malignancies are considered to increase due to improved survival rate of cancer patients, advanced diagnostic tools, and increased use of chemotherapy and radiotherapy. In Korea, several cases of triple primary malignancies were reported. However, four primary malignancies in gastro intestinal tract was rarely reported. Recently, we experienced a 70 year-old male who was diagnosed with metachronous four primary malignancies in rectum, ascending colon, stomach, and ampulla of Vater. We report this rare case of metachronous four primary malignancies with a review of literature. PMID- 19556846 TI - [A case of hepatocellular carcinoma combined with liver abscess]. AB - Hepatocellular calcinoma (HCC) is the fifth most common cancer and the third leading cause of cancer-related deaths worldwide. It is important to diagnose HCC exactly before management is attempted. But, the clinical presentations and radiologic findings of liver abscess, HCC, and metastatic tumor to the liver may be quite similar, and procedures such as serum tumor marker assay, computerized tomography, and ultrasonography of the liver cannot make a specific diagnosis. We report a case of HCC successfully diagnosed by surgery which was misconceived as liver abscess and not improved by medical treatment. PMID- 19556847 TI - [A case of autoimmune pancreatitis combined with extensive involvement of biliary tract]. AB - Autoimmune pancreatitis is a distinct disease characterized by the presence of autoantibodies and hypergammaglobulinemia, inflammation of the pancreatic parenchyma, and irregular stricture of the pancreatic duct. The involvement of distal common bile duct is frequently observed, but intrahepatic bile duct involvement is very rare, which seem to have similar feature to primary sclerosing cholangitis. We report a case of the patient with autoimmune pancreatitis combined with extensive involvement of extrahepatic and intrahepatic bile duct, which had a favorable response to steroid therapy. PMID- 19556848 TI - Enhancing immune responses to tumor-associated antigens. AB - The goal of vaccine-based cancer immunotherapy is to induce a tumor-specific immune response that ultimately reduces tumor burden. However, the immune system is often tolerant to antigens presented by the tumor, as the cancer originates from within a patient and is therefore recognized as self. This article reviews selected clinical strategies for overcoming this immune tolerance, and approaches to enhance generation of immunity to tumor-associated antigens by activating innate immunity, potentiating adaptive immunity, reducing immunosuppression, and enhancing tumor immunogenicity. Success in the field of cancer vaccines has yet to be fully realized, but intelligent choice of immunomodulators, tumor antigens and patient populations will likely lead to clinically relevant uses for cancer vaccines. PMID- 19556849 TI - Cancer cell mitochondria confer apoptosis resistance and promote metastasis. AB - Mutations in mtDNA are found in most cancers. In this study, we studied the role of cancer cell mutant mtDNA in tumorigenesis. We sequenced the entire mitochondrial genome of three different breast cancer cell lines and found that all three, MCF7, MDA-MB-231 and MDA-MB-435, contained mutations in mtDNA. MDA-MB 435 cells contained a mutation in the tRNA(Leu(CUN)) gene known to be involved in pathogenesis of mitochondrial diseases. We generated a mutant cybrid (cytoplasmic hybrid) by repopulating the recipient rho(0) (completely devoid of mtDNA) cells with donor mtDNA derived from an enucleated MDA-MB-435 breast cancer cell line. An isogenic wild-type cybrid was produced by transfer of normal mtDNA from a healthy donor. When compared to the wild type, we found that mutant mtDNA increases mitochondrial membrane potential. However, this increase in mitochondrial membrane potential was not associated with increase in reactive oxygen species (ROS) production. MtDNA mutations conferred resistance to apoptosis triggered by etoposide. Our study also revealed that mutations in mtDNA increase metastatic potential. Using a tail-vein model of metastasis in a mouse model, we show that the mutant cybrid metastatizes to the lungs and forms macrometastic foci. Additionally we found that mutations in mtDNA constitutively activate the PI3/Akt pathway that contributes to increased metastatis. Together our study demonstrates that mutant mtDNA promotes apoptotic resistance and metastasis in a mouse model. PMID- 19556850 TI - Feminizing cholinergic neurons in a male Drosophila nervous system enhances aggression. AB - Previous studies in Drosophila have demonstrated that whether flies fight like males or females can be switched by selectively manipulating genes of the sex determination hierarchy in male and female nervous systems. Here we extend these studies by demonstrating that changing the sex of cholinergic neurons in male fruit fly nervous systems via expression of the transformer gene increases the levels of aggression shown by the flies without altering the way the flies fight. Transformer manipulation in this way does not change phototaxis, geotaxis, locomotion or odor avoidance of the mutant males compared to controls. Cholinergic neurons must be feminized via this route during the late larval/early pupal stages of development to show the enhanced aggression phenotype. Other investigators have shown that this is the same time period during which sexually dimorphic patterns of behavior are specified in flies. Neurons that co-express fruitless and choline acetyl transferase are found in varying numbers within different clusters of fruitless-expressing neurons: together they make up approximately 10% of the pool of fruitless-expressing neurons in the brain and nerve cord. PMID- 19556851 TI - Argonaute-mediated translational repression (and activation). AB - MicroRNAs (miRNAs) downregulate the expression of their target genes by inducing translational repression and/or mRNA decay. Under specific conditions, miRNAs can even activate translation of their target mRNAs. These processes occur via miRNA protein complexes, or RNA-induced silencing complexes (RISCs), which contain Argonaute (Ago) subfamily protein as a core component. However, detailed mechanisms of miRNA-mediated translational regulation remain unclear. We recently reported that, in Drosophila, both of the two Ago proteins, Ago1 and Ago2, can repress translation of the target mRNAs, but by remarkably different mechanisms. Furthermore, we here show that Ago2, but not Ago1, can activate translation of the target mRNAs when they lack the poly(A) tail, suggesting that the length of poly(A) tail is an important determinant for the consequences of Ago2 function. This review focuses on how miRNAs regulate translation in light of these new findings. PMID- 19556852 TI - Induction of apoptosis by tea polyphenols mediated through mitochondrial cell death pathway in mouse skin tumors. AB - Many naturally occurring phytochemicals have shown cancer chemopreventive potential in a variety of bioassay systems. One such naturally occurring biologically active compound is tea Camellia sinensis, which is the most consumed beverage in the world after water. The most abundant and active constituents of tea are polyphenols (epigallocatechin gallate and theaflavins). In the present study, cancer chemopreventive properties of both black tea polyphenols (BTP) and green tea polyphenols (GTP) on 7,12-dimethylbenz[a]anthracene (DMBA) induced mouse skin carcinogenesis were studied. BTP and GTP treatment showed delay in onset of tumorigenesis, reduction in cumulative number of tumors and increased tumor free survival. Both BTP and GTP were found to modulate the expression of proteins involved in apoptotic pathway. Tea polyphenols treatment along with DMBA exposure resulted in upregulation of p53, and proapoptotic protein Bax, whereas enhanced expression of antiapoptotic proteins, Bcl-2 and survivin by DMBA were downregulated. Further, we showed that tea polyphenols supplementation resulted in release of cytochrome c, caspases activation, and increase in apoptotic protease activating factor and poly (ADP-ribose) polymerase cleavage as mechanism of apoptosis induction. The results also provide strong evidence that BTP is a better chemopreventive agent against skin tumorigenesis as compared to GTP at the tested dose levels. Thus, we can conclude that the polyphenolic constituents present in black tea and green tea induce mitochondrial pathway of apoptosis and hence can be used as a potential chemopreventive agents against skin cancer. PMID- 19556853 TI - Production and radioimmunoimaging of novel fully human phage display recombinant antibodies and growth inhibition of lung adenocarcinoma cell line overexpressing Prx I. AB - The Peroxiredoxin I (Prx I) is a member of the Peroxiredoxin family, which is overexpressed in many diverse tumor types and is an anti-apoptosis protein for tumor cell proliferation and survival. Therapeutic strategies targeting the Prx I may therefore be effective broad-spectrum anticancer agents. We constructed a phage display single-chain variable fragment (scFv) antibody library and sieve out the fully human, lung adenocarcinoma-sepcific monoclonal antibodies. The selection on Prx I was performed using above-mentioned lung adenocarcinoma sepcific monoclonal antibodies with high affinity to Prx I overexpressing lung adenocarcinoma cells. The candidate scFv sequences, based on enzyme-linked immunosorbent assay (ELISA) screening data, were chosen for soluble expression, and a 30 kDa band was observed on polyacrylamide gel electrophoresis as predicted. The purified antibodies were characterized by immunoblotting and showed high specificity to Prx I-overexpressing lung adenocarcinoma cells A549. Radioimmunoimaging was taken to evaluate specificity and distribution of antibodies in vivo. The radiolocalization index (RI) of tumor/serum and tumor/muscle gradually increased, reaching its peak (4.06 +/- 0.13 and 5.17 +/- 0.97, respectively) at 48 h postadministration. Single photon emission computed tomography (SPECT) imaging showed the radioactivity was aggregated in tumor locations and tumor imaging was clearly observed. The internalized scFv resulted in antibody-mediated cell apoptosis and downregulation of Prx I expression. These results demonstrate that the scFv possesses strong antitumor activity on lung adenocarcinoma and may therefore be an effective therapeutic candidate for the treatment of cancers that are dependent on Prx I for growth and survival. PMID- 19556855 TI - Prevention of skin cancer by green tea: past, present and future. PMID- 19556854 TI - Influence of Hsp70s and their regulators on yeast prion propagation. AB - Propagation of yeast prions requires normal abundance and activity of many protein chaperones. Central among them is Hsp70, a ubiquitous and essential chaperone involved in many diverse cellular processes that helps promote proper protein folding and acts as a critical component of several chaperone machines. Hsp70 is regulated by a large cohort of co-chaperones, whose effects on prions are likely mediated through Hsp70. Hsp104 is another chaperone, absent from mammalian cells, that resolubilizes proteins from aggregates. This activity, which minimally requires Hsp70 and its co-chaperone Hsp40, is essential for yeast prion replication. Although much is known about how yeast prions can be affected by altering protein chaperones, mechanistic explanations for these effects are uncertain. We discuss the variety of effects Hsp70 and its regulators have on different prions and how the effects might be due to the many ways chaperones interact with each other and with amyloid. PMID- 19556856 TI - Microparticles harbouring Sonic Hedgehog: role in angiogenesis regulation. AB - Sonic Hedgehog (Shh) is a morphogen involved in embryonic development of nervous system. Also, it has been shown that recombinant Shh can modulate angiogenesis under ischemic conditions. However, angiogenic effects of endogenous Shh have not been completely elucidated. Using small membrane-derived vesicles expressing Shh (MPs(Shh+)), we have shown that, although MPs(Shh+) decrease endothelial cell proliferation and migration, they are able to favour angiogenesis through the increase of both endothelial cell adhesion and expression of pro-angiogenenic factors. Activation of proteins implicated in cell adhesion, such as Rho A, as well as upregulation of pro-angiogenic factors were sensitive to inhibition of Shh pathway. Although whole composition of MPs(Shh+) needs to be characterized to understand potential effects of MPs(Shh+), these results highlight a new role of MPs(Shh+) in vascular pathophysiology and may have significant implication for therapy in pathologies associated with altered angiogenesis in order to re address angiogenic switch. PMID- 19556857 TI - Activation of the PI3K-Akt-mTOR signaling pathway promotes necrotic cell death via suppression of autophagy. AB - Our previous work has shown that autophagy plays a pro-survival function in two necrotic cell death models: zVAD-treated L929 cells as well as H(2)O(2)-treated Bax(-/-)Bak(-/-) mouse embryonic fibroblasts (DKO MEF). This study aims to further explore the regulatory role of autophagy in necrosis by examining the functional role of the phosphoinositide-3 kinase (PI3K)-Akt-mammalian target of rapamycin (mTOR) signaling pathway. Our initial intriguing finding was that insulin is able to promote necrotic cell death induced by zVAD and MNNG in L929 cells or by H(2)O(2) in DKO MEF cells cultured in full-growth medium. The pro necrosis function of insulin was further supported by the observations that insulin is capable of abolishing the protective effect of starvation on necrotic cell death induced by zVAD in L929 cells. Next, we demonstrated that insulin acts on the PI3K-Akt-mTOR pathway to promote necrosis as the suppression of the above pathway by either chemical inhibitors (LY294002 and rapamycin) or mTOR knockdown is able to mitigate the pro-death function of insulin. Finally, we provided evidence that the pro-death function of insulin is dependent on its inhibitory effect on autophagy, which serves as an important pro-survival function in necrosis. Taken together, here we provide compelling evidence to show that activation of the PI3K-Akt-mTOR signaling pathway can promote necrotic cell death via suppression of autophagy, at least in the necrosis models defined in our study in which autophagy serves as a pro-survival function. Data from this study not only further underscore the pro-survival function of autophagy in necrotic cell death, but also provide a novel insight into the intricate connections linking the PI3K-Akt-mTOR signaling pathway with cell death via modulation of autophagy. PMID- 19556859 TI - Mcl1 downregulation sensitizes neuroblastoma to cytotoxic chemotherapy and small molecule Bcl2-family antagonists. AB - Neuroblastoma (NB) is a common, highly lethal pediatric cancer, with treatment failures largely attributable to the emergence of chemoresistance. The pro survival Bcl2 homology (BH) proteins critically regulate apoptosis, and may represent important therapeutic targets for restoring drug sensitivity in NB. We used a human NB tumor tissue microarray to survey the expression of pro-survival BH proteins Mcl1 and Bcl2, and correlated expression to clinical prognostic factors and survival. Primary NB tumors heterogeneously expressed Mcl1 or Bcl2, with high expression correlating to high-risk phenotype. Co-expression is infrequent (11%), but correlates to reduced survival. Using RNA interference, we investigated the functional relevance of Mcl1 and Bcl2 in high-risk NB cell lines (SK-N-AS, IMR-5, NLF). Mcl1 knockdown induced apoptosis in all NB cell lines, while Bcl2 knockdown inhibited only NLF, suggesting functional heterogeneity. Finally, we determined the relevance of Mcl1 in resistance to conventional chemotherapy (etoposide, doxorubicin) and small molecule Bcl2-family antagonists (ABT-737 and AT-101). Mcl1 silencing augmented sensitivity to chemotherapeutics 2 to 300-fold, while Bcl2 silencing did not, even in Bcl2-sensitive NLF cells. Resistance to ABT-737, which targets Bcl2/-w/-x, was overcome by Mcl1 knockdown. AT-101, which also neutralizes Mcl1, had single-agent cytotoxicity, further augmented by Mcl1 knockdown. In conclusion, Mcl1 appears a predominant pro survival protein contributing to chemoresistance in NB, and Mcl1 inactivation may represent a novel therapeutic strategy. Optimization of compounds with higher Mcl1 affinity, or combination with additional Mcl1 antagonists, may enhance the clinical utility of this approach. PMID- 19556858 TI - Functional targeting of the MUC1 oncogene in human cancers. PMID- 19556861 TI - Re-evaluating membrane transport processes: new insights and appreciation of protein behavior. PMID- 19556860 TI - Atg14L and Rubicon: yin and yang of Beclin 1-mediated autophagy control. PMID- 19556862 TI - Distinct post-translational modifications regulate BK channel activity: the interplay between protein palmitoylation and phosphorylation. PMID- 19556863 TI - Chimeric IgH-TCRalpha/delta translocations in T lymphocytes mediated by RAG. AB - Translocations involving the T cell receptor alpha/delta (TCRalpha/delta) chain locus, which bring oncogenes in the proximity of the TCRalpha enhancer, are one of the hallmark features of human T cell malignancies from ataxia telangiectasia (AT) and non-AT patients. These lesions are frequently generated by the fusion of DNA breaks at the TCRalpha/delta locus to a disperse region centromeric of the immunoglobulin heavy chain (IgH) locus. Aberrant VDJ joining accounts for TCRalpha/delta associated DNA cleavage, but the molecular mechanism that leads to generation of the "oncogene partner" DNA break is unclear. Here we show that in ATM deficient primary mouse T cells, IgH/TCRalpha/delta fusions arise at a remarkably similar frequency as in human AT lymphocytes. Recombinase-activating gene (RAG) is responsible for both TCRalpha/delta as well as IgH associated breaks on chromosome 12 (Chr12), which are subject to varying degrees of chromosomal degradation. We suggest a new model for how oncogenic translocations can arise from two non-concerted physiological DSBs. PMID- 19556865 TI - Cancer research on non-small cell lung cancer in smokers and non-smokers: snapshots from the AACR annual meeting 2009. AB - The 100th annual meeting of the American Association for Cancer Research (AACR) took place in Denver on 18-22 April 2009 which brought together over 17,000 researchers from basic, translational and clinical cancer studies who exchanged and discussed recent advances in many areas of major interest. The meeting showcases the latest research trends in oncology and the related medical science. It covered a variety of different topics ranging from basic research to exploration of currently unknown pathogenesis and mechanisms for specific cancers. As the most prevalent type of lung cancer, reports on non-small cell lung cancer (NSCLC) comprised a significant part of the meeting. A number of studies looked at the molecular biology of NSCLC patients with or without smoking history. Moreover, findings on the therapeutic effects of current and investigational drugs for NSCLC were also important topics. This meeting report aims to provide the readers with a summary of the research highlights on NSCLC in smokers and non-smokers presented in the AACR annual meeting 2009. PMID- 19556864 TI - Fulvestrant treatment is associated with cholesterol plasma level reduction in hormone-receptor-positive metastatic breast cancer patients. AB - BACKGROUND: Fulvestrant is a pure anti-estrogen hormoal agent formally lacking any estrogen-agonist activity. We analyze the effect(s) of fulvestrant treatment on estrogen target systems in hormoe-sensitive advanced breast cancer patients. RESULTS: Patients received a median of five fulvestrant injections (range 3-19). We observed a partial response in one patient, disease stability in 21 and disease progression in 29 patients with a clinical benefit of 43.2% and a median time to progression of 5 [range 3-20] mo. Total cholesterol levels significantly decreased during treatment (219.8 +/- 45.3 vs. 201.4 +/- 42.1 mg/dl; p = 0.0054) together with LDL-cholesterol (129.7 +/- 41.39 vs. 112.3 +/- 37.1 mg/dl; p = 0.018). HDL-cholesterol and triglycerides did not show significant changes. Reduction of total and LDL-cholesterol was independent from last hormoal treatment or treatment duration. All coagulation indices and mean endometrial mucosa thickness value did not vary. METHODS: Fifty-one patients [median age 65 (range 48-82) y] were enrolled. All patients received previous hormoal treatments, with 90.2% receiving > or =2 courses. Last hormoal treatment was exemestane, letrozole, anastrozole and other in 30-10-7-4/51 patients respectively. Median withdrawal time was 18 d (range 3-1456). Complete fasting lipid blood profile and coagulation indices were assessed before fulvestrant administration, every 3 mo and at discontinuation time. Endometrial mucosa thickness was evaluated before fulvestrant administration and at end-study time. CONCLUSIONS: We observed a lipid lowering effect of fulvestrant possibly related to an influence on lipid metabolism by a mechanism in which a role could be played by progesterone receptor. PMID- 19556866 TI - Autophagy in food biotechnology. AB - The purpose of this review is not to explain autophagy (as clearly there is a plethora of reviews and research papers on the topic) but to provide the autophagy-savvy reader with an overview of the impact of autophagy research on a number of current topics in food biotechnology. To understand this connection, we need to remember that autophagy is, at the end of the day, a type of stress response. Since as humans we are heterotrophic eukaryotic organisms, our cells, and the cells of those organisms that we consume, use autophagy as part of the day-to-day business of living. Thus, a number of food biotechnology processes such as brewing and winemaking employ eukaryotic organisms under autophagy inducing conditions, as noted below. In addition, food spoilage processes also involve eukaryotic organisms and these processes also involve physiological aspects that impinge on autophagy. Finally, the recently introduced concept of "functional foods" introduces the possibility of engineering foodstuff for the induction or inhibition of autophagy in the consumer, with a potential promise of health benefits that merits further research. In this review, we will provide a perspective on the current literature in these three areas, their relationship to current basic research in autophagy, and their future applicative potential. PMID- 19556867 TI - An absence of stromal caveolin-1 is associated with advanced prostate cancer, metastatic disease and epithelial Akt activation. AB - Here, we examined the status of stromal Cav-1 expression in patients with benign prostatic hypertrophy (BPH), primary prostate cancers (PCa), and prostate-cancer metastases (Mets). Interestingly, an absence of stromal Cav-1 directly correlated with prostate cancer disease progression. For example, virtually all BPH samples showed abundant stromal Cav-1 immunostaining. In contrast, in a subset of patients with primary prostate cancer, the stromal levels of Cav-1 were significantly decreased, and this correlated with a high Gleason score, indicative of a worse prognosis and poor clinical outcome. Remarkably, all metastatic tumors (either from lymph node or bone) were completely negative for stromal Cav-1 staining. Thus, stromal Cav-1 expression may be considered as a new biomarker of prostate cancer disease progression and metastasis. Mechanistically, stromal Cav-1 levels were inversely correlated with the epithelial expression levels of Cav-1 and epithelial phospho-Akt. Thus, loss of stromal Cav-1 is predictive of elevated levels of epithelial Cav-1 and epithelial Akt-activation. This provides important new clinical evidence for paracrine signaling between prostate cancer epithelial cells and the tumor stromal micro-environment, especially related to disease progression and metastasis. PMID- 19556868 TI - MgAtg9 trafficking in Magnaporthe oryzae. AB - Autophagy is a vacuolar/lysosomal cytoplasmic recycling system in eukaryotic cells. ScATG9 is indispensable for autophagy in Saccharomyces cerevisiae. Here, we deleted MgATG9, the orthologue of ScATG9, via targeted gene replacement in the phytopathogenic filamentous fungus Magnaporthe oryzae, and then analyzed the cellular distribution pattern of EGFP-MgAtg9 in the Mgatg9Delta mutant. We detected an expression profile of multiple green dots in the conidial cell inoculated in rich media and in the appressoria differentiated from the conidia in H(2)O. Concurrent with the punctation, we found some fluorescent signals localized on the central vacuole of the submerged hyphae from the conidia cultured in rich media. Next, we introduced DsRed2-MgAtg8 into the Mgatg9Delta mutant expressing EGFP-MgAtg9 and observed partial overlap at multiple sites in the conidial cell, reminiscent of that in the mammalian system. Our findings further led to the postulation that the multiple sites where the two fusions colocalized tend to merge as a central structure in the conidial cell. Finally, we tested the expression of EGFP-MgAtg9 in null mutants of MgATG1, 2, 13 and 18, respectively. We speculate that MgAtg1, 2 and 18, but not MgAtg13, is required for MgAtg9 cycling through the multiple colocalization sites to its storage pools in the conidial cell of M. oryzae, and fusion of these colocalization sites into a central structure could be governed through other unidentified mechanisms. PMID- 19556869 TI - Phosphorylation of TPX2 by Plx1 enhances activation of Aurora A. AB - Entry into mitosis requires the activation of mitotic kinases, including Aurora A and Polo-like kinase 1 (Plk1). Increased levels of these kinases are frequently found associated with human cancers, and therefore it is imperative to understand the processes leading to their activation. We demonstrate that TPX2, but neither Ajuba nor Inhibitor-2, can activate Aurora A directly. Moreover, Plx1 can induce Aurora A T-loop phosphorylation indirectly in vivo during oocyte maturation. We identify Ser204 in TPX2 as a Plx1 phosphorylation site. Mutating Ser204 to alanine decreases activation of Aurora A, whereas a phosphomimetic Asp mutant exhibits enhanced activating ability. Finally, we show that phosphorylation of TPX2 with Plx1 increases its ability to activate Aurora A. Taken together, our data indicate that Plx1 promotes activation of Aurora A, most likely through TPX2. In light of the current literature, we propose a model in which Plx1 and Aurora A activate each other in a positive feedback loop. PMID- 19556870 TI - Nuclear membrane-derived autophagy, a novel process that participates in the presentation of endogenous viral antigens during HSV-1 infection. AB - Complex membrane trafficking events are involved in the regulation of antigen processing and presentation of both endogenous and exogenous antigens. While these processes were believed to involve mainly organelles along the endo/phagocytic and the biosynthetic pathways, recent studies have shown that autophagy also participates actively in both innate and adaptive immunity. We have shown recently that, in macrophages infected with the Herpes simplex type 1 virus, autophagy plays a key role in the targeting of viral proteins to hydrolytic compartments, and their processing for presentation on MHC class I molecules. This pathway involves a novel type of autophagosomes formed by coiling of the nuclear membrane where viral proteins are highly enriched. The ability to enhance the contribution of autophagy to antigen presentation in various conditions suggests that this pathway could be used to boost the immune response against viral infection and develop new vaccines. PMID- 19556871 TI - The wisdom of Weismann: epigenetic erasure mechanisms and germ line immortality. PMID- 19556872 TI - Metadherin as a link between metastasis and chemoresistance. PMID- 19556873 TI - The 3rd international conference on Influenza Vaccines for the World (IVW 2009). PMID- 19556874 TI - Transcription-blocking DNA damage in aging and longevity. PMID- 19556875 TI - To be or NOT to be demethylated. PMID- 19556877 TI - A heat-stable hepatitis B vaccine formulation. AB - The purpose of the present study was to develop a formulation of recombinant hepatitis B vaccine with improved stability at elevated temperatures. A validated in vitro antigen reactivity assay was used to measure the stability of the vaccine. The formulation development focused on modification of the interactions between the antigen and aluminum hydroxide adjuvant and subsequent optimization of the ionic aqueous environment of the adsorbed vaccine. A formulation of hepatitis B vaccine containing 40 mM histidine and 40 mM phosphate at pH 5.2 had considerably improved stability at elevated temperatures as measured by the in vitro antigen reactivity assay. The formulation exhibited 9-week stability at 55 degrees C and was subsequently shown to be stable both at 37 degrees C and at 45 degrees C for at least 6 months based on the in vitro antigen reactivity and immunogenicity in mice. The formulation comprises only excipients which have a history of safe use in approved drug products. The new vaccine formulation has the potential to be used outside the cold chain for part of its shelf life. This may improve the immunization coverage, simplify the logistics for outreach immunization, and ensure the potency of the vaccine in areas where the cold chain is insufficient. PMID- 19556876 TI - Transcription factors LSF and E2Fs: tandem cyclists driving G0 to S? AB - Cell cycle progression in mammalian cells from G(1) into S phase requires sensing and integration of multiple inputs, in order to determine whether to continue to cellular DNA replication and subsequently, to cell division. Passage to S requires transition through the restriction point, which at a molecular level consists of a bistable switch involving E2Fs and pRb family members. At the G(1)/S boundary, a number of genes essential for DNA replication and cell cycle progression are upregulated, promoting entry into S phase. Although the activating E2Fs are the most extensively characterized transcription factors driving G(1)/S expression, LSF is also a transcription factor essential for stimulating G(1)/S gene expression. A critical LSF target gene at this stage, Tyms, encodes thymidylate synthetase. In investigating how LSF is activated in a cell cycle-dependent manner, we recently identified a novel time delay mechanism for regulating its activity during G(1) progression, which is apparently independent of the E2F/pRb axis. This involves inhibition of LSF in early G(1) by two major proliferative signaling pathways: ERK and cyclin C/CDK, followed by gradual dephosphorylation during mid- to late-G(1). Whether LSF and E2F act independently or in concert to promote G(1)/S progression remains to be determined. PMID- 19556878 TI - Adjuvants and delivery technologies. PMID- 19556880 TI - Vascular endothelial growth factor receptor-3 promotes breast cancer cell proliferation, motility and survival in vitro and tumor formation in vivo. AB - Vascular endothelial growth factor receptor-3 is a receptor tyrosine kinase that is overexpressed in some human carcinomas, but its role in tumorigenesis has not been fully elucidated. We examined VEGFR-3 expression in normal, nonneoplastic and early stage malignant breast tissues and have shown that VEGFR-3 upregulation in breast cancer preceded tumor cell invasion, suggesting that VEGFR-3 may function as a survival signal. We characterized the biological effects of VEGFR-3 over-expression in human breast cancer cells based on two approaches: gain of function by overexpressing VEGFR-3 in MCF-7 breast cancer cells and loss of function by RNAi-mediated silencing of VEGFR-3 in MCF-7-VEGFR-3 and BT474 cells. VEGFR-3 overexpression increased cellular proliferation by 40% when MCF7-VEGFR-3 cells were compared to parental MCF7 cells, and proliferation was reduced by more than 40% when endogenous VEGFR-3 was downregulated in BT474 cells. VEGFR-3 overexpression promoted a three-fold increase in motility and invasion and both motility and invasion were inhibited by downregulation of VEGFR-3. Furthermore, VEGFR-3 overexpression promoted cellular survival under stress conditions induced by staurosporine treatment and led to anchorage-independent growth. VEGFR-3 overexpression dramatically increased tumor formation in both hormone-dependent and independent xenograft models. With estrogen stimulation, MCF7-VEGFR-3 xenografts were ten times larger than control xenografts. Finally, downregulation of VEGFR-3 expression in both xenograft model cell lines led to a significant reduction of tumor growth. For the first time, we have demonstrated that VEGFR-3 overexpression promotes breast cancer cell proliferation, motility, survival, anchorage-independent growth and tumorogenicity in the absence of ligand expression. PMID- 19556881 TI - The oncogenic potential of a bystander. PMID- 19556879 TI - Network architecture of signaling from uncoupled helicase-polymerase to cell cycle checkpoints and trans-lesion DNA synthesis. AB - When replication is blocked by a template lesion or polymerase inhibitor while helicase continues unwinding the DNA, single stranded DNA (ssDNA) accumulates and becomes coated with RPA, which then initiates signals via PCNA mono ubiquitination to activate trans-lesion polymerases and via ATR and Chk1 to inhibit Cdk2-dependent cell cycle progression. The signals are conveyed by way of a complex network of molecular interactions. To clarify those complexities, we have constructed a molecular interaction map (MIM) using a novel hierarchical assembly procedure. Molecules were arranged on the map in hierarchical levels according to interaction step distance from the DNA region of stalled replication. The hierarchical MIM allows us to disentangle the network's interlocking pathways and loops and to suggest functionally significant features of network architecture. The MIM shows how parallel pathways and multiple feedback loops can provide failsafe and robust switch-like responses to replication stress. Within the central level of hierarchy ATR and Claspin together appear to function as a nexus that conveys signals from many sources to many destinations. We noted a division of labor between those two molecules, separating enzymatic and structural roles. In addition, the network architecture disclosed by the hierarchical map, suggested a speculative model for how molecular crowding and the granular localization of network components in the cell nucleus can facilitate function. PMID- 19556882 TI - Dipeptidyl peptidase 2 is an essential survival factor in the regulation of cell quiescence. AB - Most cells in the body are in a resting state and undergo cell cycle progression only upon growth factor stimulation or activation. while much research on proliferation and activation has been performed, very little about signals that maintain quiescent cells in G(0) is known, preventing cell cycle entry or apoptosis. In this study, the pathways of apoptosis induction in quiescent peripheral blood cells and fibroblasts mediated by inhibition or downregulation of Dipeptidyl Peptidase 2 (DPP2) have been explored. A decrease in DPP2 activity was found to cause resting cells to exit from G(0), accompanied by a decrease in p130, p27(Kip1) and p21(Cip1) protein levels. In addition, DPP2-inhibited or downregulated cells exhibit an increase in early G(1)/S progressors, with increases in the levels of retinoblastoma (pRb), p107 and cyclin D proteins. Furthermore, decrease of DPP2 activity leads to an increase in c-Myc and a decrease in Bcl-2, two events that have been associated with apoptosis induction. This apoptosis by DPP2 downregulation is prevented in p53(-/-) cells or by ectopic expression of proteins that suppress p53 or c-Myc activity. Thus, DPP2 is essential for maintaining lymphocytes and fibroblasts in G(0), and its inhibition results in apoptosis mediated by induction of c-Myc and p53. PMID- 19556883 TI - Phylogenetic taxonomy in Drosophila. AB - The genus Drosophila is one of the best-studied model systems in modern biology, with twelve fully sequenced genomes available. In spite of the large number of genetic and genomic resources, little is known concerning the phylogenetic relationships, ecology and evolutionary history of all but a few species. Recent molecular systematic studies have shown that this genus is comprised of at least three independent lineages and that several other genera are actually imbedded within Drosophila. This genus accounts for over 2,000 described, and many more undescribed, species. While some Drosophila researchers are advocating dividing this genus into three or more separate genera, others favor maintaining Drosophila as a single large genus. With the recent sequencing of the genomes of multiple Drosophila species and their expanding use in comparative biology, it is critical that the Drosophila research community understands the taxonomic framework underlying the naming and relationships of these species. The subdivision of this genus has significant biological implications, ranging from the accurate annotation of single genes to understanding how ecological adaptations have occurred over the history of the group. PMID- 19556884 TI - The prognostic role of Beclin 1 protein expression in high-grade gliomas. AB - High-grade gliomas (HGG) have a poor outcome, however, prognostic subgroups of patients may be individuated by some clinico-biological parameters. It was recently demonstrated that the main response of HGG to therapy is autophagic death. Autophagy is involved in tumor suppression, and is defective in HGG, in which we previously found an underexpression of beclin 1 autophagic gene protein product. Underexpression of Beclin 1 protein has been correlated to poor patient outcome in other tumor types. In this paper, the prognostic role of Beclin 1 expression in HGG patients was investigated. We first evaluated the tumor cell cytoplasmic expression of Beclin 1 protein (BPCE), in a sample of 76 HGG by immunohistochemistry, and compared it with cell proliferation and apoptosis. We found high BPCE score positively correlated with apoptosis, and negatively with cell proliferation (p < 0.05). We then correlated BPCE score with survival and other prognostic parameters (histological grading, MGMT gene methylation status, age, patient performance status according to the Karnofski classification (KPS), extent of surgery, radiation therapy (RT) modality, temozolomide chemotherapy (TMZ CHT), and optimal/suboptimal post-surgical treatment). Forty-seven (61.8%) and twenty-nine (38.2%) patients showed high and low BPCE scores, respectively. BPCE showed statistically significant correlations with survival both at the univariate (p = 0.03) and multivariate analysis (p = 0.037). High BPCE was also positively correlated with high KPS values (p = 0.023), and with the accomplishment of an optimal postoperative therapy (p = 0.037). Furthermore, among patients showing a MGMT methylated gene, survival was significantly higher in cases with a higher BPCE score. BPCE score might be added to pathological evaluation of HGG for prognostic purposes. PMID- 19556885 TI - Analysis of DRAM-related proteins reveals evolutionarily conserved and divergent roles in the control of autophagy. AB - Autophagy is a membrane-trafficking process that serves to deliver cytoplasmic proteins and organelles to the lysosome for degradation. The process is genetically defined and many of the factors involved are conserved from yeast to man. Recently, a number of new autophagy regulators have been defined, including the Damage-Regulated Autophagy Modulator (DRAM), which is a lysosomal protein that links autophagy and the tumor suppressor, p53. We describe here analysis of DRAM-related proteins which reveals evolutionary conservation and divergence of DRAM's role in autophagy. We report that humans have 5 other proteins that show significant homology to DRAM. The closest of these, which we have termed DRAM2, displays 45% identity and 67% conservation when compared to DRAM. Interestingly, although similar to DRAM in terms of homology, DRAM2 is different from DRAM as it not induced by p53 or p73. DRAM2 is also a lysosomal protein, but again unlike DRAM its overexpression does not modulate autophagy. In contrast to humans, the Drosophila genome only encodes one DRAM-like protein, which is approximately equal in similarity to human DRAM and DRAM2. This questions, therefore, whether DRAM function is conserved from fly to man or whether DRAM's capacity to regulate autophagy has evolved in higher eukaryotes. Expression of DmDRAM, however, clearly revealed an ability to modulate autophagy. This points, therefore, to a conserved role of DRAM in this process and that additional human proteins have more recently evolved which, while potentially sharing some similarities with DRAM, may not be as intrinsically connected to autophagy regulation. PMID- 19556886 TI - Leaving the midline: how Robo receptors regulate the guidance of post-crossing spinal commissural axons. AB - In the developing nervous system, pathfinding axons navigate through a series of intermediate targets in order to form synaptic connections. Vertebrate spinal commissural axons extend toward and across the floor plate (FP), a key intermediate target located at the ventral midline (VM). Subsequently, post crossing commissural axons grow either alongside or significant distances away from the floor plate (FP), but never re-cross the VM. Consistent with this behavior, post-crossing commissural axons lose responsiveness to the FP associated chemoattractants, Netrin-1 and SHH, and gain responsiveness to Slits, which are potent midline repellents, in vitro. In addition, the results of several in vivo studies suggest that the upregulation of Slit-binding repulsive Robo receptors, Robo1/2, alters the responsiveness of decussated commissural axons to midline guidance cues. Nevertheless, in vertebrates, it is unclear whether Robo1/2 are the sole or major repellent receptors responsible for driving these commissural axons away from the VM and preventing their re-entry into the FP. We recently re-visited these issues in the chick spinal cord by assessing the consequences of manipulating Robo expression on commissural axons in ovo. Our findings suggest that, at least in chick embryos, the upregulation of repulsive Robos on post-crossing axons alters the responsiveness of these axons to midline repellents and facilitates their expulsion from, but is not likely to have a significant role in preventing their re-entry into the VM. PMID- 19556887 TI - Disparities in preschool immunization coverage associated with maternal age. AB - Associations between maternal age and preschool immunization coverage are unclear. This study aimed to determine if maternal age is associated with preschool immunization coverage and the importance of maternal age compared with other factors affecting vaccination coverage. Data from the 2001-2003 National Immunization Survey (NIS) were used to estimate vaccine coverage. Children were considered up-to-date (UTD) if they received > or =4 doses of DTaP, > or =3 doses of polio, > or =1 doses of MMR, > or =3 doses of Hib and > or =3 doses of Hep B. Bivariate and multivariate relationships between UTD coverage and maternal, child and household factors were evaluated. Classification tree analysis assessed complex interactions between maternal, child and household factors associated with UTD coverage and isolated the most important factors in predicting UTD coverage. UTD coverage was significantly associated with maternal age: coverage increased as maternal age increased. Coverage among children with 17 year old mothers was 64%; coverage among children of mothers 17-26 years old increased by 16.3% overall (approximately 1.8% per year). After 26 years of age, coverage did not increase significantly as maternal age increased. The relationship between maternal age and UTD coverage remained statistically significant after adjusting for a broad range of maternal, child and household factors. Classification tree analysis suggested that maternal age is the most important factor associated with vaccine coverage. More research is needed to determine the reasons for underimmunization of children born to young mothers. PMID- 19556888 TI - Challenges in developing a pediatric RSV vaccine. AB - RSV bronchiolitis is the leading cause of infant hospitalization in industrialized countries. There is an unmet need to prevent RSV lower respiratory tract infection in young infants. Although many vaccinology approaches, including live attenuated, viral and bacterial vectored and adjuvanted subunit vaccines have been evaluated in rodent and primate models there is currently no approved RSV vaccine. A vaccine candidate for RSV-naive infants must provide immunogenicity in the presence of maternally acquired antibodies, avoid enhanced disease and have minimal reactogenicity. Because live RSV infection does not potentiate for enhanced disease and elicits systemic and mucosal immune responses, live RSV vaccine candidates are currently preferred. Two live attenuated RSV vaccine candidates, rA2cpts248/404/1030/DeltaSH, a temperature sensitive RSV with a deletion of the SH gene, and rb/h PIV3/RSV F2 which has RSV F vectored into a bovine/human chimeric parainfluenza type 3 genome, have recently advanced into clinical studies. PMID- 19556890 TI - MACC1 controls Met: what a difference an Sp1 site makes. PMID- 19556889 TI - Many colorectal cancers are "flat" clonal expansions. AB - Population geneticists can reconstruct the ancestries of macroscopic populations from polymorphisms in present day individuals. For example, the migration "out of Africa" is recorded in human genome variation in different parts of the world. Here we apply this approach to human colorectal cancer cell populations and polymorphic passenger methylation patterns. By sampling molecular variation from different parts of the same cancer, it should be possible to infer how individual tumors grow because recent clonal expansions should be less diverse than older expansions. Average diversity was different between cancers implying that some cancers are older clonal expansions than others. For individual cancers, methylation pattern diversity was relatively uniform throughout the tumor (right versus left side, superficial versus invasive), which is more consistent with a single, uniform or "flat" clonal expansion than with stepwise sequential progression. Many colorectal cancers appear to invade and expand early, but subsequently stall. Epiallele diversity within individual small cancer gland fragments was high and more consistent with frequent rather than extremely rare cancer stem cells (CSCs). These studies suggest that many human colorectal cancers are relatively old uniform clonal expansions, that cancer cell populations contain frequent long-lived CSC lineages, and that some passenger methylation patterns record somatic cell ancestry. PMID- 19556891 TI - Modeling the molecular consequences of unbalanced translocations in cancer: lessons from acute lymphoblastic leukemia. AB - Chromosomal rearrangements are recurrent findings in human cancer and result in aberrant restructuring of the genome. The majority of known fusion genes are the consequence of reciprocal (balanced) translocations. However, most translocations described in human cancer are unbalanced, suggesting that other cancer genes remain to be identified. Historically, it was assumed that these unbalanced rearrangements affected gene function through the loss or gain of chromosomal material. However, emerging data supports direct disruption of genes located at or close to the unbalanced translocation breakpoints. New approaches are required for the identification of those gene loci underlying unbalanced translocations in cancer, as traditional methods have had limited success. This review focuses on one such strategy, using traditional and innovative molecular technologies to characterize breakpoint heterogeneity within a series of acute lymphoblastic leukemia (ALL) patients with dicentric chromosomes. This approach has shown that in ALL, specific gene loci can be targeted by heterogeneous translocation breakpoints involving multiple partner chromosomes. Carcinomas have a high proportion of unbalanced rearrangements and relatively few significant genes have been identified. The application of the same strategy to their analysis will lead to the discovery of novel cancer genes and improve our understanding of the genetic basis of tumorigenesis. PMID- 19556892 TI - Regulated expression of cofilin and the consequent regulation of p27(kip1) are essential for G(1) phase progression. AB - Cofilin, a ubiquitously expressed actin binding protein, is responsible for the formation of the actin cytoskeleton and is indispensable for cell cycle control. However, the association between cofilin expression and the cell cycle remains to be elucidated. In this study, we found that the expression level of cofilin upregulated in G(1) phase-arrested confluent cells, while knockdown of cofilin expression by small interference RNA (siRNA) in these cells led to a reduction in the population of G(1) cells. To investigate the role of cofilin in the control of G(1) phase progression, a tet-on gene expression system was introduced to overexpress different concentrations of cofilin in cells. The results showed that G(1) phase progression was blocked following induction of exogenous cofilin. A survey of the cell cycle proteins controlling the G(1) phase progression revealed that the cyclin-dependent kinase inhibitor (CKI) p27(kip1) was the primary molecule induced by overexpressed cofilin in a time and dose dependent manner. Upregulated p27(kip1) repressed phosphorylation of the retinoblastoma protein (Rb) mediated by cyclin D1/CDK4 activity. Conversely, siRNA against p27(kip1) expression in the cofilin overexpressing cells released the G(1) phase arrest. Furthermore, we found that overexpression of cofilin led to induction of p27(kip1) gene promoter transactivation using luciferase reporter gene assay. This effect was associated with increase of p27(kip1) mRNA transiently. In addition, inhibition of threonine-187 phosphorylation of p27(kip1) protein for ubiquitinyl-proteasomal mediated degradation was also involved in upregulation of p27(kip1). These data suggest that cofilin expression and its regulation of p27(kip1) expression is important for the control of G(1) phase progression. PMID- 19556893 TI - The active form of the metabolic sensor: AMP-activated protein kinase (AMPK) directly binds the mitotic apparatus and travels from centrosomes to the spindle midzone during mitosis and cytokinesis. AB - The metabolic rheostat AMP-activated protein kinase (AMPK) is unexpectedly required for proper cell division and faithful chromosomal segregation during mitosis. Although it is conceptually attractive to assume that AMPK-interpreted microenvironmental bioenergetics may strictly engage cell's energy status, cell grow, and cell division to avoid that energy stresses trigger cell death, the ultimate framework of AMPK activity towards chromosomal and cytoskeletal mitotic regulation is a question that remains unanswered. We herein reveal that the active form of the alpha-catalytic AMPK subunit (P-AMPKalpha(Thr172))-but not its total form (AMPKalpha)-transiently associates with several mitotic structures including centrosomes, spindle poles, the central spindle midzone and the midbody throughout all of the mitotic stages and cytokinesis in human cancer-derived epithelial cells. At prophase, P-AMPKalpha(Thr172) associates with the two asters of microtubules that begin to nucleate from mature centrosomes. The overlapping localization of P-AMPKalpha(Thr172) with the mitotic centrosomal Aurora-A kinase is also apparent on the microtubules near the spindle poles in metaphase and in early anaphase. This Aurora A-like centrosomal localization of P AMPKalpha(Thr172) cannot be detected following chromatid separation following anaphase-telophase transition. Rather, toward the end of anaphase and in telophase P-AMPKalpha(Thr172) reactivity exhibited a similar but not identical localization to that occupied by the bona fide chromosomal passenger proteins INCENCP and Aurora-B. This localization of P-AMPKalpha(Thr172) at the central spindle and midbody persisted during the furrowing process and, at the completion of telophase, staining of P-AMPKalpha(Thr172) as doublet was apparent on either side of the midbody within the intercellular cytokinetic bridge. An identical mitotic geography of P-AMPKalpha(Thr172) was observed in cancer cells lacking the AMPK kinase LKB1, in non-cancerous human epithelial cells, and in mouse fibroblasts. The active form of AMPKalpha bound to the mitotic apparatus may physically tether the bioenergetic state of a cell to the four-dimensional regulation of the chromosomal and cytoskeletal mitotic events, thus suggesting a putative cytokinetic suppressor function. In this newly discovered scenario, we suggest a primordial mitotic role for the alpha catalytic AMPK subunit in the eukaryotic evolutionary process as it may ensure, at the cell level, an exquisite coordination between sensing of energy resources and the fundamental biological process of genome division. PMID- 19556894 TI - The cellular prion protein and its role in Alzheimer disease. AB - The cellular prion protein (PrP(C)) is a membrane-bound glycoprotein especially abundant in the central nervous system (CNS). The scrapie prion protein (PrP(Sc,) also termed prions) is responsible of transmissible spongiform encephalopathies (TSE), a group of neurodegenerative diseases which affect humans and other mammal species, although the presence of PrP(C) is needed for the establishment and further evolution of prions. The present work compares the expression and localization of PrP(C) between healthy human brains and those suffering from Alzheimer disease (AD). In both situations we have observed a rostrocaudal decrease in the amount of PrP(C) within the CNS, both by immunoblotting and immunohistochemistry techniques. PrP(C) is higher expressed in our control brains than in AD cases. There was a neuronal loss and astogliosis in our AD cases. There was a tendency of a lesser expression of PrP(C) in AD cases than in healthy ones. And in AD cases, the intensity of the expression of the unglycosylated band is higher than the di- and monoglycosylated bands. With regards to amyloid plaques, those present in AD cases were positively labeled for PrP(C), a result which is further supported by the presence of PrP(C) in the amyloid plaques of a transgenic line of mice mimicking AD. The work was done according to Helsinki Declaration of 1975, and approved by the Ethics Committee of the Faculty of Medicine of the University of Navarre. PMID- 19556896 TI - The novel regulator of metazoan development SAYP organizes a nuclear coactivator supercomplex. AB - SAYP is a dual-function transcription coactivator of RNA polymerase II. It is a metazoan-specific factor with regulated expression that is apparently involved in signaling pathways controlling normal development. In Drosophila, SAYP is maternally loaded into the embryo, participates in cell cycle synchronization in early syncytial embryos, and is indispensible for early embryogenesis. SAYP is abundant in many embryonic tissues and imaginal discs in larvae and is crucial for oogenesis in adults. PHF10 is a mammalian homologue of SAYP whose expression is confined to certain tissues in adults. The molecular mechanism of the SAYP function is related to the conserved domain SAY, which assembles a nuclear supercomplex BTFly consisting of Brahma and TFIID coactivators. We suggest that nuclear supercomplexes may be important means of gene-specific regulation of transcription during development. PMID- 19556897 TI - REGgamma/PA28gamma proteasome activator interacts with PML and Chk2 and affects PML nuclear bodies number. AB - REGgamma (also called PA28gamma or PSME3) is a proteasome activator involved in the degradation of several proteins that regulate cell cycle and transcription. Recently, we demonstrated that this protein has a role also in the maintenance of chromosomal stability and in the response to spindle damaging agents. Here we report for the first time that REGgamma interacts with the promyelocytic leukemia protein (PML), accumulates in PML nuclear bodies (PML-NBs), but it does not play any role in normal or arsenic-induced PML degradation. However, REGgamma seems to regulate PML-NBs number, since its deficiency causes an increase in PML-NBs, which can be overcome by increased levels of SUMO1, and its overexpression has the opposite effect. We additionally found that REGgamma interacts with the DNA damage checkpoint kinase Chk2, whose presence is necessary for the increase of PML-NBs induced by REGgamma deficiency, and that REGgamma depletion resulted in a partial restoration of PML-NBs in APL derived cells. Altogether, these results underline a new role for REGgamma in the control and regulation of PML subnuclear structures. PMID- 19556895 TI - Microtubule network asymmetry in motile cells: role of Golgi-derived array. AB - Cell migration requires polarization of the cell into the leading edge and the trailing edge. Microtubules (MTs) are indispensable for polarized cell migration in the majority of cell types. To support cell polarity, MT network has to be functionally and structurally asymmetric. How is this asymmetry achieved? In interphase cells, MTs form a dynamic system radiating from a centrosome-based MT organizing center (MTOC) to the cell edges. Symmetry of this radial array can be broken according to four general principles. Asymmetry occurs due to differential modulation of MT dynamics, relocation of existing MTs within a cell, adding an asymmetric nucleation site, and/or repositioning of a symmetric nucleation site to one side of a cell. Combinations of these asymmetry regulation principles result in a variety of asymmetric MT networks typical for diverse motile cell types. Importantly, an asymmetric MT array is formed at a non-conventional MT nucleation site, the Golgi. Here, we emphasize the contribution of this array to the asymmetry of MT network. PMID- 19556898 TI - Recent advances using anti-CTLA-4 for the treatment of melanoma. AB - Metastatic melanoma is a disease associated with poor prognosis, with a median survival reported to range from 6 to 9 months. Patients who are not candidates for surgical resection have an even worse expected survival. This is largely due to the lack of effective chemotherapeutic regimens and has led to the investigation of alternative treatment strategies including immunotherapy. Although melanoma is felt to be an immunogenic tumor and has been associated with the development of spontaneous tumor-specific immune responses in patients, the implementation of vaccine-based treatment has had limited success. Because the administration of a melanoma-specific vaccine alone has not been sufficient to generate robust and reproducible clinical responses, investigators are currently pursuing additional methods to augment antimelanoma immune responses by optimizing T-cell activation. T-cell activation requires both antigen presentation to the T-cell receptor and a second signal mediated by CD80 and CD86 on antigen-presenting cells and CD28 on the T cell. Ligand binding to CD28 on the T-cell surface leads to T-cell proliferation and expression of activating cytokines such as interleukin-2. Cytotoxic T-lymphocyte antigen-4 (CTLA-4), an inhibitory protein expressed on T cells, competes for the same ligands as CD28 and modulates T-cell activation. Because CTLA-4 has a significantly higher binding efficiency than CD28, CTLA-4 is critical in maintaining immune tolerance to self-antigens and may also limit responses to tumor antigens and vaccine therapy. CTLA-4 blockade either alone or in combination with melanoma-specific vaccines has been explored as a potential means to treat advanced stage melanoma. In this article, we review the spectrum of clinical trials involving CTLA-4 blockade and also review recent correlative studies attempting to elucidate the potential mechanisms by which CTLA-4 blockade achieves its therapeutic effects. PMID- 19556899 TI - Late morbidity after successful treatment of children with cancer. AB - Over the last 4 decades, there has been a tremendous improvement in survival of children diagnosed with cancer, with 5-year survival rates now averaging 80%. The rapidly growing population of childhood cancer survivors creates an obligation to understand the health and well being of these individuals. Use of cancer therapy at an early age can produce a large burden of morbidity, as demonstrated quite conclusively by the fact that approximately two thirds of these survivors will experience at least one late effect, and approximately one third will experience a late effect, that is, severe or life threatening. Long-term complications in childhood cancer survivors, such as impairment in growth and development, neurocognitive dysfunction, cardiopulmonary compromise, endocrine dysfunction, renal impairment, gastrointestinal dysfunction, musculoskeletal sequelae, and second cancers, are related not only to the specific therapy used, but may also be determined by individual host characteristics. This review provides an update of the known late effects observed in childhood cancer survivors to provide the rationale for evaluation of specific long-term problems in this growing population of individuals at risk for chronic health conditions. PMID- 19556900 TI - Foreword: peritoneal surface malignancies: a real challenge for surgeons. PMID- 19556901 TI - From the guest editors: introduction: progress in the management of carcinomatosis. PMID- 19556902 TI - Imaging of peritoneal carcinomatosis. AB - Imaging studies are crucial in the evaluation of patients with suspected or known peritoneal cancerous dissemination. Despite the major progress that has occurred in radiological technology in the last few years, adequate and early detection of peritoneal surface disease remains a challenge. Improvements in spatial resolution are still insufficient to detect small volume peritoneal implants, often resulting in an underestimation of peritoneal disease burden, as assessed at subsequent surgical exploration. Cytoreductive surgery combined with perioperative intraperitoneal chemotherapy has provided unprecedented results in the management of peritoneal-based neoplasms, provided that a complete (adequate) cytoreduction is achieved. Diagnostic imaging tests are used to select patients who may benefit from this combined treatment by ruling out extraperitoneal involvement and signs of unresectable peritoneal disease. Furthermore, a careful assessment of the disease distribution within the peritoneal cavity, guided by a deep knowledge of the disease's clinical and biological behavior helps in planning the surgical procedure. Close interaction and cooperation between surgeons and radiologists is of utmost importance in this regard, and dedicated, motivated radiologists are required. Contrast-enhanced, multidetector computed tomography scan remains the standard imaging modality in the assessment of peritoneal carcinomatosis. Magnetic resonance imaging may offer complementary valuable data. Positron emission tomography (PET) has a more limited role, its main indication being the detection of unsuspected extraperitoneal involvement in nonmucinous neoplasms. PMID- 19556903 TI - Laparoscopy in the management of peritoneal carcinomatosis. AB - The use of videolaparoscopy in the assessment of peritoneal carcinomatosis extent is now universally accepted. This procedure allows us to define with certainty the origin of the neoplasm, the peritoneal cancer index (PCI), the involvement of the small bowel and its mesentery, the feasibility of surgery and the index of attainable cytoreduction, the evaluation of an eventual multiorgan resection, and finally results in no mortality and very low morbidity.The indications for laparoscopy are as follows: staging of the carcinomatosis already diagnosed with imaging (CT scan and MRI), staging of carcinomatosis of dubious origin (biopsy), restaging after neoadjuvant chemotherapy, restaging during follow-up in the case of dubious imaging, and restaging after adjuvant chemotherapy.Open (Hasson) technique has always been used in the introduction of the first trocar, and the changing position of the surgical bed allows for the evaluation of all the abdominal quadrants, limiting viscerolysis to the essential minimum to avoid iatrogenic lesions. Associating the intraoperative ultrasound has allowed us to reduce understaging of lesions at the depth of the diaphragm, of hepatic metastases and neoplastic masses at the pancreatic tail, and of the omental retrocavity.In all the cases in which diagnostic laparoscopy was followed by peritonectomy, we found a good correlation between open surgery data and the laparoscopic PCI. We excluded patients from peritonectomy if the staging laparoscopy showed a significant involvement of the small bowel or mesentery.We used videolaparoscopy to stage 197 cases of peritoneal carcinomatosis and achieved full laparoscopic PCI assessment in 196 of 197 (99.49%) cases, whereas only 4 of 197 (2.03%) cases were understaged before the routine use of laparoscopic ultrasound. Four complications were observed: 2 cases (1.02%) involved an infection of the trocar insertion site, which was treated with antibiotic therapy, and 2 cases (1.02%) involved diaphragm perforation and intraoperative bleeding, respectively, both resolved with videolaparoscopic technique. Two trocars were sufficient in 184 of 197 cases. There was no mortality and no port site metastasis.More recently, we have used videolaparoscopic surgery in the treatment of neoplastic ascites that did not respond to chemotherapy for palliative purposes, which resulted in the total disappearance of the ascites. It is now possible, in light of acquired experience, to evaluate with precision the indications, the technique, and the limits of the method both in the assessment of the carcinomatosis extent and in its palliation. In 28 cases of neoplastic ascites nonrespondent to chemotherapy, we were able to implement fully laparoscopic hyperthermic chemotherapy for the palliative treatment of the ascites, with total disappearance of it in all cases. The l-hyperthermic intraperineal chemotherapy was carried out at 42 degrees C for 90 minutes with 1.5% dextrose solution as a carrier. The chemotherapy solution was cisplatin and doxorubicin, or mitomycin, depending on the type of primary tumor. The drains were left in place and were removed when profuse drainage ceased.Ascites were controlled in all treated cases. A computed tomography scan performed in follow-up showed a small, clinically undetectable, fluid accumulation in the pelvis of 1 patient. Neither mortality nor morbidity was observed in connection with the procedure. PMID- 19556904 TI - Morbidity and mortality with cytoreductive surgery and intraperitoneal chemotherapy: the importance of a learning curve. AB - Evidence for the efficacy of cytoreductive surgery, combined with hyperthermic intraperitoneal chemotherapy (HIPEC), in the treatment of peritoneal carcinomatosis is accumulating. Many centers around the world now have considerable experience of the complex techniques required to achieve complete cytoreduction with the administration of HIPEC. Procedure-related morbidity ranges from 12% to 67.6% and mortality from 0% to 9% in recent studies of cytoreductive surgery and HIPEC for pseudomyxoma peritonei. A number of specialized centers have studied the factors that influence perioperative complications and mortality and have demonstrated impressive reductions in morbidity and mortality over time. However, for this treatment to be accepted as standard of care, teams undertaking this treatment strategy must aim to minimize morbidity and mortality by learning from the experience of established centers and using the "global learning curve." PMID- 19556905 TI - Cytoreductive surgery plus intraperitoneal chemohyperthermia in patients with colorectal cancer at high risk for local-regional recurrence. AB - This article was written to define the situations in which early second-look surgery with a combined treatment should be indicated in patients at high risk of developing peritoneal carcinomatosis (PC). Through a review of the literature, this is a definition of the second-look concept and of the different groups of patients at risk, in different situations (after resection of the primary, after initial cytoreductive surgery plus hyperthermic intraperitoneal chemotherapy, and after the discovery of isolated carcinoembryonic antigen elevation or isolated peritoneal uptake on positron emission tomography scan). Systematic second-look surgery for early treatment of asymptomatic PC is probably beneficial in patients presenting a high risk of developing PC after resection of their primary. The benefit seems considerably lower for the other groups of high-risk patients. PMID- 19556906 TI - Surgical techniques in visceral resection and peritonectomy procedures. AB - Peritoneal surface malignancies may now be approached with curative intent using a strategy of cytoreductive surgery combined with intraperitoneal chemotherapy. Complete cytoreduction to tumor nodules less than 2.5 mm is considered critical to the efficacy of the local chemotherapy and achieving complete cytoreduction impacts long-term survival. However, this procedure results in significant morbidity with frequent surgical complications. Meticulous technique must be used to limit anastomotic complications, operative time, and blood loss. This review summarizes the major issues surrounding visceral resection and peritonectomy during cytoreductive surgery. Problem areas, which merit special attention, are described, and recommendations are made with regard to surgical technique. PMID- 19556907 TI - Long-term results of cytoreduction and HIPEC followed by systemic chemotherapy. AB - Cytoreduction followed by hyperthermic intraperitoneal chemotherapy is a treatment option for peritoneal surface malignancies in The Netherlands. This treatment has been available for more than 10 years. Therefore, long-term results on survival and quality of life can now be studied. With these results, the true long-term benefits of this new management strategy can be determined. PMID- 19556908 TI - Pharmacokinetics and pharmacodynamics of perioperative cancer chemotherapy in peritoneal surface malignancy. AB - The peritoneal surface remains an important failure site for patients with gastrointestinal and gynecologic malignancies. During the last 2 decades, novel therapeutic approaches, combining cytoreductive surgery with intraoperative intracavitary and intravenous chemotherapy, have emerged for peritoneal carcinomatosis patients. This has resulted in remarkable clinical successes in contrast with prior failures. Although further clinical data from phase II and III trials supporting this combined treatment protocols are necessary, an optimalization of the wide variety of different perioperative cancer chemotherapy protocols used in these treatment regimens is equally important. To this date, a clear understanding of the pharmacology of perioperative chemotherapy is still lacking. The efficacy of intraperitoneal cancer chemotherapy protocols is governed as much by nonpharmacokinetic variables (tumor nodule size, density, vascularity, interstitial fluid pressure, and binding) as by the pharmacokinetic variables (dose, volume, duration, pressure, and carrier solution). Our recent data support the importance of the tumor nodule as the most meaningful pharmacologic end point. Timing of perioperative intravenous chemotherapy may substantially influence the pharmacokinetics. This review aims to clarify the pharmacokinetic and pharmacodynamic data currently available regarding the intraperitoneal delivery of cancer chemotherapy agents in patients with peritoneal carcinomatosis. PMID- 19556909 TI - Epithelial appendiceal neoplasms. AB - The appendiceal malignancies usually arise within a mucocele. The tumor within this structure can be minimally aggressive or of an invasive character. If a low grade appendiceal malignancy is removed intact, recurrence does not occur. If rupture of the wall of the mucocele occurs with either low-grade or high-grade disease, the epithelial cells within will disseminate to the peritoneal surfaces. Sometimes a second-look surgery must be performed to determine if intraperitoneal spread of mucus and cancer cells has occurred. If the diagnosis of peritoneal dissemination of an appendiceal malignancy has been established, a new treatment with curative intent is indicated. Cytoreductive surgery is combined with perioperative intraperitoneal chemotherapy as a comprehensive management plan. In approximately 900 patients treated at the Washington Cancer Institute, the quantitative prognostic indicators for appendiceal cancer with peritoneal dissemination have been determined. Patients with a complete cytoreduction and low-grade tumor have an 80% survival at 20 years; with high-grade tumors, the survival drops to approximately 45%. The extent of malignancy present within the abdomen by the peritoneal cancer index has a significant impact on survival for both high-grade and low-grade disease. The most important indicator is the completeness of cytoreduction; for both high-grade and low-grade disease, all patients with an incomplete cytoreduction have died by 10 years. The presence versus absence of lymph nodal metastases on survival have minimal significance (P = 0.0495). These results of treatment can be achieved with a 19% incidence of severe complications and a 2% mortality. PMID- 19556910 TI - Patient selection for a curative approach to carcinomatosis. AB - There is an increasing evidence showing that in selected patients with peritoneal carcinomatosis cytoreductive surgery and hyperthermic intraperitoneal chemotherapy may improve survival. Adequate patient selection is crucial to obtain a complete macroscopic cytoreduction, a leading predictor of patient outcome. However, selection is a very difficult process and is associated with a significant learning curve. Many selection criteria have to be assessed in each patient: performance status, comorbiditites, response to previous chemotherapies, histology grading, and presence of extra-abdominal or liver metastases, small bowel involvement, and tumor volume assessed by the peritoneal cancer index. All these factors have to be discussed interdisciplinary and with the patient to create an individualized treatment strategy. It is difficult to decide the relative importance of each selection criteria. However, completeness of cytoreduction, tumor volume, and histology grading are most important in many multivariate analysis independent prognostic factors. For appropriate selected patients with peritoneal carcinomatosis arising from appendiceal and colon cancer, cytoreductive surgery and hyperthermic intraperitoneal chemotherapy should be considered standard of care. PMID- 19556911 TI - Management of peritoneal carcinomatosis from colorectal cancer: current state of practice. AB - Peritoneal carcinomatosis is a common manifestation of colorectal cancer and has traditionally been regarded as a terminal disease with a short median survival. Over the 2 past decades, a new local-regional therapeutic approach combining cytoreductive surgery with intraperitoneal chemohyperthermia (hyperthermic intraperitoneal chemotherapy) has evolved. Because of its significant but acceptable morbidity and mortality and cost, careful patient selection is needed for this comprehensive management plan. Quantitative prognostic indicators are necessary as an assessment of a patient's eligibility for combined treatment. In large phase II studies, international registries, and in one phase III study, this therapeutic strategy demonstrated promising survival results with possibility of cure. In all studies, complete cytoreduction with no visible disease remaining is a requirement for long-term benefit. Further collaboration between peritoneal surface malignancy treatment centers may help to standardize indications and techniques for hyperthermic intraperitoneal chemotherapy and peritonectomy. The development and validation of novel protocols and guidelines will allow surgeons and oncologists who discover colorectal carcinomatosis to treat these patients effectively. PMID- 19556912 TI - Technology of hyperthermic intraperitoneal chemotherapy in the United States, Europe, China, Japan, and Korea. AB - Significant improvements in the understanding of the biologic behavior of peritoneal surface malignancies in addition to the combination of peritonectomy procedures that allow complete eradication of macroscopic peritoneal disease and hyperthermic intraperitoneal chemotherapy (HIPEC) at the time of surgery, directed at residual microscopic disease, have change the therapeutic strategy from a palliative approach to a curative intent in a selected group of patients with peritoneal carcinomatosis.The rationale for adding HIPEC is supported by the strong pharmacological advantage over systemic therapy. Because of the peritoneal plasma barrier, intraperitoneal administration of chemotherapy results in intraperitoneal levels that are 20 to 1000 times higher than plasma levels. The chemotherapy not only directly destroys tumor cells, but also eliminates viable platelets, neutrophils, and monocytes from the peritoneal cavity. This diminishes the promotion of tumor growth associated with the wound healing process. In addition, combining the intraperitoneal chemotherapy with hyperthermia has several advantages. Heat by itself has more toxicity for cancerous tissue than for normal tissue, and this predominant effect on cancer increases as the vascularity of the malignancy decreases. Also, hyperthermia increases the penetration of chemotherapy into tissues. As tissues soften in response to heat, the elevated interstitial pressure of a tumor mass may decrease and allow improved drug penetration. Lastly, and probably most important, heat increases the cytotoxicity of selected chemotherapy agents. This synergism occurs only at the interface of heat and body tissue at the peritoneal surface.However, despite the wider acceptance to combine extensive cytoreductive surgery with intraoperative intraperitoneal heated chemotherapy, the specifics of the HIPEC administration continue to lack uniformity. The most recent consensus statement issued by the Peritoneal Surface Oncology Group International after the 2006 meeting in Milan concluded that the debate on the best method to deliver HIPEC is still open, and as a group, we declared that there is no sufficient evidence in the literature confirming the superiority of one technique over the other in terms of outcome, morbidity, and safety to the personnel in the operating room. PMID- 19556913 TI - Intrahepatic cholangiocarcinoma: a worthy challenge. PMID- 19556915 TI - Editor's note: the power of words. PMID- 19556914 TI - Retrospective analysis of histopathologic prognostic factors after hepatectomy for intrahepatic cholangiocarcinoma. AB - BACKGROUND: The aim of this study was to investigate histopathologic prognostic factors in patients with intrahepatic cholangiocarcinoma (ICC) whose tumors were resected to determine the optimal surgical strategies. METHODS: One hundred and two ICC patients who underwent laparotomy from July 1998 to December 2000 were followed up successfully. Histopathologic variables were selected for univariate and multivariate analyses to evaluate their influence on the outcome. RESULTS: The 1-, 3-, and 5-year survival rates after surgery were 56.9%, 25.5%, and 16.9%, respectively. The average survival duration was 21.91 +/- 20.17 months. In univariate analysis, the presence of lymph node (LN) metastasis, number of LNs with metastases, presence of intrahepatic metastasis, curative resection, and TNM stage were significant risk factors for survival. Multivariate analysis revealed that intrahepatic metastasis, noncurative resection, and TNM stage IVa were independent prognostic factors. CONCLUSIONS: The histopathologic characteristics of intrahepatic metastasis were closely related to poor prognosis in ICC patients. Extensive hepatectomy with LN dissection may offer the only chance for long-term survival in patients with ICC. PMID- 19556916 TI - A systematic review of the effect of moderate intensity exercise on function and disease progression in amyotrophic lateral sclerosis. AB - BACKGROUND AND PURPOSE: Amyotrophic lateral sclerosis (ALS) is an idiopathic disease of adults affecting upper and lower motor neurons. In one to four years, progressive weakness, spasticity, and respiratory insufficiency compromise independence and survival. Current medical treatment is limited to medication and supportive care. The benefit and harm of moderate physical exercise are controversial. This review examined current research related to moderate exercise for maintaining independence without accelerating disease progression in persons with ALS. METHODS: An evidence-based search was conducted using keywords alone and in combination (ALS, exercise, Lou Gehrig's disease, physical therapy) to search PubMed, PEDro, Hooked on Evidence, Ovid, and Cochrane databases. Human and animal models were included and graded on level of evidence and strength of recommendations for developing guidelines to practice. A secondary reviewer evaluated all selected studies, and statistics were calculated. RESULTS: The search yielded the following nine studies: four small clinical studies, one clinical systematic review, and four randomized, controlled trials based on animal models. In human studies, there were small to moderate effect sizes supporting the benefit of moderate exercise in persons with early-stage ALS, with no adverse affects on disease progression or survival time. In transgenic mice with superoxide dismutase-1 ALS, moderate exercise most often had a moderate effect size for increasing life span. DISCUSSION AND CONCLUSION: Large randomized clinical trials are needed to develop specific exercise guidelines. However, evidence suggests that moderate exercise is not associated with adverse outcomes in persons with early-stage ALS. Moderate exercise programs can be safely adapted to abilities, interests, specific response to exercise, accessibility, and family support. PMID- 19556917 TI - Short-term effects of posture-assisted step training on rapid step initiation in Parkinson's disease. AB - BACKGROUND AND PURPOSE: Anticipatory postural adjustments (APAs) for lateral weight transfer and stability precede and accompany gait initiation. Individuals with Parkinson's disease (PD) show altered APA characteristics with delays in initiating stepping that may reflect impaired interactions between posture and locomotion. The purpose of this study was to determine the short-term effects of a single session of repetitive robotic assistance training with the APA on rapid step initiation in individuals with PD in the medications "on" state and healthy control individuals. Ground reaction forces and step kinematics were recorded. METHODS: Subjects first performed baseline trials of unassisted self-paced rapid forward stepping. Next, a training acquisition series involved 50 trials with a lateral pull applied to the pelvis by a robotic system to assist with the early phase of the APA during stepping. To assess potential retention effects of training, unassisted stepping trials were evaluated immediately after acquisition trials (immediate retention) and one week later (one-week retention). RESULTS: Overall, the subjects with PD had a longer APA duration (P < 0.03), and longer first step duration (P < 0.04) than the healthy control individuals. Compared with baseline, APA duration was shorter (P < 0.001) and step onset time became earlier (P < 0.001) for acquisition trials but these effects were not retained. Step duration, which became shorter (P < 0.001) during the late acquisition trials (P = 0.002), demonstrated immediate retention (P < 0.001) and one-week retention (P < 0.001). CONCLUSION: Posture-assisted training, affecting the interaction between posture and locomotion, may have therapeutic potential for improving movement performance in individuals with PD. PMID- 19556919 TI - Gait in individuals with chronic hemiparesis: one-year follow-up of the effects of a neuroprosthesis that ameliorates foot drop. AB - PURPOSE: Foot drop is a common cause of gait impairment in individuals with hemiparesis. The objective of this study was to determine the short-term and long term effects on functional gait of a neuroprosthesis that provides functional electrical stimulation (FES) to correct footdrop. METHODS: Sixteen individuals (mean age, 55.0 +/- 14.6 years) with chronic hemiparesis (mean years since insult, 5.3 +/- 4.8) were assessed immediately before receiving the neuroprosthesis and two months and one year after using the device. Both follow up assessments were conducted with FES operating; the one-year follow-up also assessed gait without FES. Outcome measures were gait velocity on an even surface during a 10-m walk, during a six-minute walk, over obstacles, and over a carpet. Foot switches used during the six-minute walk determined stance time, stride time variability, and gait asymmetry. The effect of time and the one-year carryover effect on gait without FES were determined, with significance set at P = 0.05. RESULTS: The neuroprosthesis had a significant short-term and long-term effect on most measured variables. Thus, mean 10-m walk gait velocity improved in two months from 0.67 m/sec to 0.86 m/sec and continued to increase to 1.06 m/sec by the one-year assessment. A carryover effect on gait velocity and stance time without FES were also noted. The 10-m gait velocity without the neuroprosthesis increased by 23.8% by the one-year follow-up assessment. CONCLUSIONS: The neuroprosthesis appears to improve gait performance, and gains were further enhanced over time and carried over to gait even without FES. These results suggest orthotic and therapeutic effects on footdrop after prolonged use of the neuroprosthesis. PMID- 19556918 TI - Relationships and responsiveness of six upper extremity function tests during the first six months of recovery after stroke. AB - BACKGROUND: Knowing how clinical tests are related to each other and whether tests are responsive to change are critical to evidence-based practice and sound research. OBJECTIVE: To determine (1) relationships between six clinical tests of upper extremity function and (2) responsiveness for each test during the first six months after stroke. METHODS: Grip strength test, pinch strength test, Action Research Arm Test, Jebsen-Taylor Hand Function Test, Nine-Hole Peg Test, and the Stroke Impact Scale-Hand domain were administered to 33 subjects within one, three, and six months after stroke. Spearman correlations were used to analyze relationships between tests. Responsiveness was calculated using the single population effect size method. RESULTS: All tests were correlated with each other with absolute r-values ranging from 0.54 to 0.92 at one month, 0.57 to 0.97 at three months, and 0.41 to 0.97 at six months. All tests were at least moderately responsive to change, with effect sizes ranging from 0.50 to 1.02 from one to three months, and 0.56 to 0.86 (d) from one to six months. CONCLUSIONS: Our data can assist clinicians and researchers in making decisions to use specific tests for measuring upper extremity function in people with hemiparesis in the first six months of recovery. PMID- 19556920 TI - The recovery of running ability in an adolescent male after traumatic brain injury: a case study. AB - BACKGROUND AND PURPOSE: The purpose of this case study was to document outcomes after a rehabilitation program in an adolescent male after traumatic brain injury. Three years after sustaining an injury in a skiing accident, a 17-year old boy participated in a rehabilitation program with the goal of acquiring the ability to run one mile with his peers. On initial evaluation, the individual had significant left lower extremity weakness, impaired standing balance, limited endurance, and running limitations. He was able to run 10 m wearing a plastic ankle-foot orthosis on the left side but required supervision for safety. METHODS: The intervention included strength training once weekly for 17 weeks, body weight-supported, treadmill-based locomotor training once weekly for 15 weeks followed by a combination of overground locomotor training and strengthening exercise once weekly for six weeks. OUTCOMES: After the intervention, muscle strength of the lower extremities increased and the individual was able to run one mile independently. The quality of his running improved, with better mechanics to absorb forces at impact during the absorption phase and increased lower extremity extension during the propulsion phase. DISCUSSION: A rehabilitation program consisting of strengthening and locomotor training improved running speed, quality, and endurance in an adolescent male after traumatic brain injury. He was able to progress to a less restrictive carbon fiber brace as a result of gains in lower extremity strength. This change in ability allowed him to participate in physical education by running on a track and playing softball with his peers. PMID- 19556921 TI - JNPT adapting to a time of "uncertainty" with "certainty". PMID- 19556925 TI - Risk factors for repeat syphilis in men who have sex with men, San Francisco. AB - BACKGROUND: Syphilis incidence has increased dramatically in the United States since 2000, occurring primarily among men who have sex with men (MSM) and disproportionately affecting those with human immunodeficiency virus (HIV) infection. The continued increases in syphilis rates among MSM signals the need for enhanced prevention methods. We undertook a study to examine the rate of repeat syphilis infection among MSM in San Francisco and to identify risk factors associated with syphilis reinfection that may inform additional prevention strategies. METHODS: We developed a retrospective cohort of all cases of primary, secondary, and early latent syphilis among MSM diagnosed in San Francisco in 2001 and 2002. We evaluated data through the end of 2003 to determine all cases of syphilis reinfection, defined as a new infection that occurred within 1 year after prior syphilis infection and treatment. RESULTS: We found that 6.7% (42/624) of cases had a repeat syphilis infection within 1 year. HIV infection was associated with an increased risk of repeat infection (OR = 4.7; CI, 1.8 12.0). No differences in age, race, number of period sex partners, illicit substance use, or partner meeting venues were observed between cases with and without repeat infection. CONCLUSIONS: Our study revealed that HIV-infected MSM with syphilis represent an at-risk group for repeat syphilis infection. Targeting increased screening and risk reduction interventions to HIV-infected MSM in care could reduce the overall incidence of syphilis among MSM. PMID- 19556927 TI - Antimicrobial susceptibility profile of Neisseria gonorrhoeae isolated from patients attending a STD facility in Maputo, Mozambique. AB - OBJECTIVE: To ascertain the effectiveness of kanamycin for the treatment of gonorrhoea in Maputo, Mozambique. METHODS & DESIGN: A cross-sectional study design was employed. Urethral and cervical specimens were collected for the isolation of Neisseria gonorrhoeae from patients attending Centro de Saude do Porto. Antimicrobial susceptibilities were determined for kanamycin, spectinomycin, ciprofloxacin, ceftriaxone, cefixime, tetracycline and penicillin. RESULTS: Twenty-two (40%) Neisseria gonorrhoeae isolates were intermediate and 4(7%) were resistant to kanamycin; 42(77%) displayed high level resistance to tetracycline (MIC > or = 16 mg/L); 34 (65%) were penicillinase producers, and 52 (95%) had spectinomycin MICs of 64 mg/L. All isolates were susceptible to ciprofloxacin (MIC < or = 0.06 mg/L), ceftriaxone (MIC < or = 0.015 mg/L) and cefixime (MIC < or = 0.015 mg/L). CONCLUSION: The observations underscore the need for broader susceptibility surveillance studies to elucidate the pattern and extent of drug resistance in Mozambique. A review of the current treatment guidelines for genital discharge syndrome is warranted. PMID- 19556928 TI - Upstairs and downstairs: socio-economic and gender interactions in herpes simplex virus type 2 seroprevalence in australia. AB - BACKGROUND: This study investigates socio-economic differentials in herpes simplex virus type 2 (HSV-2) seroprevalence in Australian men and women using individual and geographic measures of socio-economic status. METHODS: HSV-2 seropositivity among men and women aged over 25 years was investigated by levels of individual and area-based measures of socio-economic status (SES) in a series of Poisson regression models, variously adjusting for age, country of birth, marital status, indigenous status, and urban/rural residence as potential confounders. Serum and socio-demographics were collected during 1999 and 2000 in a population-based Australia-wide prevalence survey. RESULTS: HSV-2 seroprevalence was significantly lower in areas of low SES than in high SES areas among both men (P for trend <0.001) and women (P for trend = 0.004) for all ages. A similar pattern was evident for individual education level for men with lower rates of HSV-2 in respondents with lower educational achievement (relative risk = 0.77, 95% CI 0.61-0.97, P = 0.024). In contrast, HSV-2 prevalence was higher for women with lower individual levels of education for all ages (relative risk = 1.22, 95% CI 1.04-1.44, P = 0.017). Analyses stratifying HSV-2 prevalence for individual education level by area-based SES showed the highest prevalence of HSV 2 in women with the lowest education level residing in the highest SES areas. This pattern was not evident in men, with a greater concordance between individual and area-based SES. CONCLUSION: HSV-2 seroprevalence is not consistently distributed across individual and area measures of SES, suggesting that upward and downward mixing between social strata in men and women is an important mode of HSV-2 transmission. PMID- 19556926 TI - The association between oral contraceptives, depot-medroxyprogesterone acetate, and trichomoniasis. AB - BACKGROUND: Hormonal contraception use by women may increase the risk of acquiring certain sexually transmitted infections. We explored the effect of hormonal contraceptive use, specifically oral contraception (OC), and depot medroxyprogesterone acetate (DMPA) on Trichomonas vaginalis infections in women. METHODS: We examined data from a prospective case-control study of women with trichomoniasis and noninfected female patients recruited from 3 public sexually transmitted disease clinics. Women with positive wet mount microscopy or T. vaginalis culture results were classified as having trichomoniasis. Participants underwent physical examinations, sexually transmitted infections testing and completed questionnaires which included information about demographics, sexual behavior, douching and contraceptive use. We assessed the association between hormonal contraceptives and trichomoniasis using bivariable and multivariable analysis and estimated exposure odds ratios (ORs) and adjusted odds ratios (aORs) with 95% confidence intervals (CIs). RESULTS: We identified 427 women with trichomoniasis and 144 uninfected women who had information reported about contraception use. Compared with nonhormonal contraceptive use, OC use was negatively associated with trichomoniasis in bivariable analysis (OR: 0.5; 95% CI: 0.3-0.8). This association was no longer statistically significant after adjusting for demographic variables, douching and condom use (aOR: 0.9; 95% CI: 0.5-1.6). Use of DMPA, compared with nonhormonal contraceptive use, was not associated with trichomoniasis in bivariable or multivariable analyses (OR: 1.0, 95% CI: 0.5-2.1; aOR = 1.4, 95% CI: 0.6-3.4, respectively). CONCLUSIONS: Although OC use appeared to have a protective effect in the bivariable analysis, the hormonal contraceptives OC and DMPA were not associated with T. vaginalis infection after adjustment for other factors. PMID- 19556930 TI - Limitations of the dye-based method for determining vaginal applicator use in microbicide trials. AB - BACKGROUND: A dye-based method for determining applicator usage in microbicide trials has been developed to assess whether applicators have been exposed to vaginal fluid. Our objective was to evaluate this method on polypropylene HTI applicators that are being widely used in several effectiveness trials of microbicides. METHODS: Study participants enrolled in a clinical trial assessing SPL7013 (VivaGel) inserted gel intravaginally twice daily for 14 days and returned used and unused applicators. Before staining, smears were prepared from each participant-inserted applicator, Gram stained and assessed independently for the presence of vaginal cells and bacteria. Of the 169 participant-inserted applicators, 168 (99%) had vaginal cells identified by Gram stain. PARTICIPANT: Inserted applicators were stained with a 0.05% FD & C blue dye No. 1 solution and compared with 70 inserted positive control applicators and 70 unused negative control applicators. Intravaginally inserted applicators should stain turquoise, whereas unused applicators should not retain any stain. The individual responsible for labeling and preparing the applicators did not serve as an evaluator. RESULTS: : Under optimized conditions, the sensitivity and specificity ranged from 81% to 95% and 86% to 93%, respectively for single use and unused applicators. The dye-based method was only 47% to 77% sensitive for participant inserted applicators obtained from women using gel twice daily. CONCLUSION: The dye test for HTI polypropylene applicators had a sensitivity of 47% to 95%, depending on the evaluator and whether gel was present in the vagina. The sensitivity was decreased with multiple gel applications. The dye-based method cannot be recommended for HTI polypropylene applicators to monitor product adherence. PMID- 19556929 TI - A summary of preclinical topical microbicide rectal safety and efficacy evaluations in a pigtailed macaque model. AB - BACKGROUND: There is widespread recognition of the potential promise of vaginal microbicides as a tool to combat global human immunodeficiency virus/acquired immunodeficiency syndrome and sexually transmitted infections epidemics, and candidate product development has maintained a rapid pace in recent years; however, rectal microbicide development has received less attention. As it is likely that commercial products developed for vaginal use will also be used rectally, there is a clear need to assess the safety and efficacy of candidate microbicide products specifically in the rectal compartment. METHODS: We have developed a standardized protocol for preclinical rectal safety and (chlamydial) efficacy assessment of topical microbicide candidates in a nonhuman primate model. We evaluated a total of 12 test compounds for rectal safety (via rectal pH, microflora, and rectal lavage) and 1 compound for efficacy against rectal chlamydial infection. RESULTS: In this article, we describe our methods in detail and summarize our results, particularly noting the ability of our model to distinguish products with deleterious effects on the rectal environment. We also outline the specific criteria used to recommend products move into preclinical rectal efficacy trials or be recommended for reformulation to the product developer. In summary, we observed significant adverse effects in 2 products. The single product that underwent efficacy evaluation was not observed to be protective against rectal chlamydial infection. CONCLUSIONS: A preclinical safety and efficacy model is critical to promoting rectal microbicide development, which will ultimately offer a significant opportunity for intervention in the global HIV/AIDS epidemic. PMID- 19556931 TI - Patterns of treatment of external genital warts in Australian sexual health clinics. AB - BACKGROUND: External genital warts are a common sexually transmitted viral disease. We describe the patterns of treatment for initial presentations of external genital warts (EGWs) in Australian sexual health centers. METHODS: This was a retrospective audit of 489 case notes from consecutive individuals who presented with a new diagnosis of EGWs to 1 of 5 major sexual health clinics in Australia. Eligibility criteria were consecutive patients aged 18 to 45 years inclusively, presenting with first ever episode of EGWs from January 1, 2004. Exclusion criteria were patients who were immunocompromised, including HIV infection, or enrollment in a treatment study for EGWs. RESULTS: The median age at presentation of women was 23.2 years and of men 26.8 years. One quarter (n = 127) of patients had another sexually transmitted infection diagnosed at presentation. Nearly half of the patients (n = 224) presented only once for treatment. Most often, patients were treated with a monotherapy (n = 382/489; 78%), usually cryotherapy (257; 53%). Staff applied treatment in 361 (74%) cases. There was wide variation across sites, possibly reflecting local policies and budgets. We found no difference in wart resolution (n = 292; 60%) by initial treatment chosen. CONCLUSIONS: The diagnosis and treatment of genital warts constitute a sizable proportion of clinical visits to the audited sexual health services and require a large input of staff time to manage, including the application of topical treatments. Our results help complete the picture of the burden of EGWs on Australian sexual health centers before the introduction of the HPV vaccine. PMID- 19556932 TI - Twenty-year trends in the incidence and prevalence of diagnosed anogenital warts in Canada. AB - BACKGROUND: A vaccine has recently been licensed in many countries that protects against the human papillomavirus types 6, 11, 16, and 18. Types 6 and 11 account for approximately 90% of anogenital warts (AGWs). We describe the 20-year trends in the incidence and prevalence of AGWs in Manitoba, Canada. METHODS: We used linked population-based hospital and physician databases for Manitoba for 1984 to 2004. Cases were identified using tariff (billing) and ICD codes. A case was considered to be incident if it was preceded by a 12-month interval free period of AGWs care. Otherwise, it was deemed to be prevalent. An episode was considered over once a 12-month interval had elapsed without an AGW claim. RESULTS: Approximately 25,000 Manitobans were diagnosed with AGWs between 1985 and 2004. The annual age-standardized incidence rates peaked in 1992 (men, 149.9/100,000; women 170.8/100,000). In recent years, the rates have been increasing again, particularly for men. The male:female incidence rate ratio increased from 0.76 in 1985 to 1.25 in 2004. The highest incidence rate tended to be in those aged 20 to 24 years. Trends in prevalence were similar. Prevalence in 2004 was 165.2/100,000 for men and 128.4/100,000 for women. CONCLUSIONS: These population-based findings suggest that AGWs are a substantial burden to Manitobans and that their pattern has changed over time, with incidence and prevalence becoming higher in men than women. Monitoring the future trends in AGWs will provide an early marker of the effectiveness and duration of protection of human papillomavirus vaccination at a population level. PMID- 19556933 TI - Effect of acculturation on the acceptability of potential microbicides and sexual risk-taking. AB - BACKGROUND: The objective was to determine the acceptability and use patterns of potential microbicides among African American (AA), acculturated Hispanic (AH), and less acculturated Hispanic (LAH) women. We measured baseline sexual risk taking and the likelihood of behavioral change, given effective microbicides. METHODS: Interview of 506 Mexican-American and AA women, all of whom have a sexually transmitted infection enrolled in Project Sexual Awareness for Everyone. RESULTS: The 3 groups reported similarly high acceptance of potential microbicides (76%-83% P = 0.24). LAHs were most likely to report they would use microbicides covertly (P = 0.03). Given the possibility of effective microbicides, AHs were consistently more likely to report risk disinhibition. AHs, as compared to LAHs and AAs, respectively, were most likely to report that they would not use condoms, (53% vs. 33% vs. 30% P <0.001), would have a 1-night stand (18% vs. 8% vs. 6% P = 0.02), or would have sex with humans before they got to know them (18% vs. 8% vs. 6% P = 0.01). AHs were also most likely to say they would or probably would change from baseline safe sexual practices to unsafe sexual behaviors if potential microbicides were available. Age was controlled for in the analysis as AHs were younger than AAs and LAHs. CONCLUSIONS: Future microbicides were acceptable among this at risk cohort. Acculturation was a predictor of risk disinhibition and should be considered when tailoring sexually transmitted infection prevention messages, given the advent of effective microbicides. PMID- 19556934 TI - Estimates of the direct cost per case and overall burden of trichomoniasis for the employer-sponsored privately insured women population in the United States, 2001 to 2005. AB - BACKGROUND: Little is known about the direct medical cost and overall burden of trichomoniasis among women in the United States. METHODS: We extracted insurance claims for trichomoniasis for 2001 to 2005 from the MEDSTAT MarketScan database using International Classification of Diseases, ninth revision codes. The analysis was restricted to outpatient care and prescription drug claims for women in 4 age categories; under 15, 15 to 24, 25 to 34, and 35 to 64. We used Current Procedures Terminology codes to analyze diagnostic methodologies. All costs were adjusted to 2005 US dollars. RESULTS: The average outpatient and prescription drug costs per episode for all ages were 97 dollars and 9 dollars, respectively. The resulting average total cost per episode was 101 dollars (about 50% did not have drug costs). Average total cost among women aged 15 to 24 years (120 dollars) was significantly (P < 0.01) higher than all other age categories. The estimated annual economic burden was 6.8 million dollars among privately insured women and 18.9 million dollars among all women from the United States. The incidence rate for female enrollees (all ages) having claims was 91 per 100,000 enrollees. Incidence rates were highest for women aged 25 to 29 years (185 per 100,000), followed by women aged 20 to 24 years (166 per 100,000). The most common diagnostic procedure seemed to be wet mount, but nonspecificity of Current Procedures Terminology codes inhibited the analysis of diagnostic methodologies. CONCLUSION: The estimated economic burden was highest among reproductive age women (15-34 years). Our estimated economic burden represents a lower-bound estimate because it was based on direct medical costs only. PMID- 19556935 TI - Diagnostic methodologies for chlamydia screening in females aged 15 to 25 years from private insurance claims data in the United States, 2001 to 2005. PMID- 19556936 TI - Changes in community socioeconomic status and racial distribution associated with gonorrhea rates: an analysis at the community level. AB - BACKGROUND: Gonorrhea persists despite nationwide disease control measures. Research has not focused on the longitudinal impact of community characteristics on gonorrhea. The authors assessed the association between community demographic and socioeconomic changes and gonorrhea rates. METHODS: Gonorrhea incident cases reported in 1992-1993 and 2001-2002 were aggregated at the census tract level in New York State exclusive of New York City and then matched to the census data. Census tract characteristics were obtained from the 1990 and 2000 decennial censuses and adjusted for changes in geographic boundaries. RESULTS: Gonorrhea incidence declined considerably between 2 study periods (1992-1993 and 2001-2002) among urban census tracts, but gradually increased in suburbs and rural areas. Changes in community socioeconomic status (SES) were significantly associated with change in gonorrhea rates (e.g., gonorrhea rate ratio given a 5% increase in household poverty rate = 1.08; 95% confidence interval: 1.02-1.14), independent of changes in community demographics. Increases in gonorrhea rates related to the proportion of non-Hispanic black population within urban census tracts persisted after multiple SES variables were controlled (rate ratio = 1.20; 95% confidence interval: 1.16-1.24). DISCUSSION: This study found that temporal changes in community SES and demographic characteristics were associated with changes in gonorrhea rates. PMID- 19556937 TI - Knowledge and practice among healthcare providers in rural Vietnam regarding sexually transmitted infections. AB - OBJECTIVES: To assess knowledge and reported practice regarding sexually transmitted infections (STI) among healthcare providers in rural Vietnam and to examine background characteristics possibly associated with knowledge and practice. METHODS: A cross-sectional study using a self-completed questionnaire was carried out in 2006 among 465 healthcare providers in rural Vietnam. The questionnaire included questions on providers' characteristics, STI knowledge, and case scenarios of 4 common STI syndromes. Correct answer was scored 1, "do not know" or incorrect answer was scored 0. Linear and logistic regressions were applied. RESULTS: Diseases considered as STI were gonorrhea and syphilis by 83% of the respondents, 70% believed partner treatment necessary for bacterial vaginosis or candidiasis cases. Sharing clothes/food or kissing was commonly mentioned as transmission routes (60%). Median scores of knowledge and reported practice were 29 (range: 0-50) and 2 (range: 0-20), respectively. Among the respondents, 34% had a knowledge score of less than 25 and 78% had a practice score of less than 10. Characteristics predicting higher level of knowledge were being a medical doctor, assistant medical doctor, midwife, or serving STI patients. Characteristics predicting higher level of practice were serving STI patients, being a midwife or female provider, and having participated in STI or reproductive tract infection training courses. Respondents who reported treating STI patients had a higher level of knowledge and reported practice than the others. PMID- 19556938 TI - Dammarane-type triterpene saponins from the flowers of Panax notoginseng. AB - Four new dammarane-type triterpene saponins named floranotoginsenosides A (1), B (2), C (3) and D (4), together with five known triterpene saponins, were isolated from the flowers of Panax notoginseng. Their structures were elucidated on the basis of spectral and chemical evidence. PMID- 19556939 TI - A medical food for Alzheimer's disease. PMID- 19556940 TI - Biennial [corrected] IV zoledronic acid (Reclast) for prevention of osteoporosis. PMID- 19556941 TI - Lacosamide for epilepsy. PMID- 19556942 TI - Drugs for treatment of chronic heart failure. PMID- 19556943 TI - The historical development of psychiatry in Serbia. AB - The authors present the development of the concept of mental disease and treatment in Serbian medicine. Serbian medieval medicine did not acknowledge fortune telling, sorcery, the use of amulets and magical rituals and formulas. These progressive concepts were confirmed by the Church and the Serbian state in what is known as Dusan's Code. The Historical data on the establishment of the first psychiatric hospital in the Balkans "Home for the Unsound of Mind" at Guberevac, Belgrade, in 1861 and its founders is reviewed. After World War I, in 1923, the Faculty of Medicine was established in Belgrade to which the coryphaei of Serbian medicine educated in Europe, mostly in France and Germany, flocked and that same year the Psychiatry Clinic of the Faculty of Medicine in Belgrade was set up. Its first seat was on the premises of the Mental Hospital in Belgrade, and it became a training base and laid the foundations of the future Neuropsychiatry Clinic in Belgrade, which in time evolved into the nursery of psychiatric professionals for all of Serbia. The most important data on the further development of psychiatry up to date are presented. PMID- 19556944 TI - Neuropsychological profile of delusional disorder. AB - BACKGROUND: Previous studies concerned with neuropsychological aspect of delusions, were mainly focused on specific forms of this disorder, such as Cotard or Capgras type of delusions. Comparatively small numbers of investigations were concerned with cognitive deficiencies accompanying the delusions. The substance of this study includes the detection of neuropsychological dysfunctions in patients with delusional disorder, and tracing of these cognitive distortions to appropriate brain regions. SUBJECTS AND METHODS: The investigation is designed as a comparative study. Inpatients with delusion are compared with normal subjects from the aspect of the following cognitive functions: attention, memory, visuospatial and visuoconstruction organization, executive ability, verbal divergent thinking. RESULTS: Attention, memory (verbal modality) and psychomotor skill tasks are most susceptible to delusional effects. CONCLUSION: The neuropsychological profile of patients with delusional disorder includes impediment of complex attention modalities. From this primary disorder, there also stems a disorder of verbal memory in the sense of reduced recognition. These cognitive distortions suggest a dysfunction of the anterior regions of the cerebrum, mainly of the prefrontal and sinistral temporal regions. PMID- 19556945 TI - Comparative analysis of soft neurological signs in positive and negative subtype of schizophrenia. AB - BACKGROUND: The objective of the study was to investigate neurological deficit in schizophrenia and to compare soft neurological signs in positive and negative subtypes of schizophrenia. SUBJECTS AND METHODS: 66 patients with schizophrenia were evaluated with the Positive and Negative Syndrome Scale to classify the subtype of schizophrenia: positive subtype (36 patients) and negative subtype (30 patients), all of which were entering into remission. To examine the neurological soft signs we compared scores on the Neurological Evaluation Scale (NES) for positive and negative subtype. RESULTS: The negative subtype of schizophrenia showed significantly higher neurological soft signs in comparison to the positive subtype, with reduced functioning in the sensory integration and motor coordination subscale as well as the other subscale. CONCLUSION: The main finding in this study indicates that patients with schizophrenia have neurological impairment, and that the negative subtype has significantly higher neurological impairment than the positive subtype. The results further support the significance of the soft neurological signs as a possible marker of different subtypes of schizophrenia. PMID- 19556946 TI - Long-term efficacy of electroconvulsive therapy combined with different antipsychotic drugs in previously resistant schizophrenia. AB - BACKGROUND: In treatment-resistant schizophrenia a combination of ECT with antipsychotics has been reported to have superior outcomes compared to other strategies, however the results were inconsistent. We investigated the long-term effects of the combination of unilateral, non-dominant hemisphere ECT with three antipsychotics. SUBJECTS AND METHODS: The clinical study was a naturalistic, prospective, open-labeled, active-controlled study in adult outpatients of both genders suffering from treatment-resistant schizophrenia with a follow up of 2 years. The patients received sulpiride (n=17, 100-400mg/day, PO), risperidone (n=26, 2-8 mg/day, PO) or olanzapine (n=27, 5-10mg/day, PO). Unilateral ECT was applied in 1 unit (0.5A, 0.8 mS) in six single applications, once a week and further according to the clinical need, in fortnight steps. Clinical efficacy was established using the PANSS and CGI psychometric scales. RESULTS: According to the results, the most effective treatment mode was olanzapine plus ECT, then risperidone plus ECT, while sulpiride plus ECT had lower clinical efficacy. Olanzapine plus ECT was significantly superior in all scale scores vs sulpiride plus ECT, as well as risperidone plus ECT except for PANSS-P (t=1.85, p>0.05). During the study, 38 of 70 patients were withdrawn due to treatment failure (n=21), side effects (n=6) and non-compliance (n=11). CONCLUSION: The combination of novel antipsychotics and ECT can be used safely and effectively in treatment resistant schizophrenia. PMID- 19556947 TI - Lamotrigine versus lithium augmentation of antidepressant therapy in treatment resistant depression: efficacy and tolerability. AB - BACKGROUND: Mood stabilizer augmentation of standard antidepressant drugs has been shown to be effective in treatment-resistant depression. Despite the reported high overall efficacy, lithium has been relatively underused in recent years. Lamotrigine, a novel anticonvulsant recently recognized as a mood stabilizer, seems to have putative antidepressive properties. The aim of the study was to investigate lamotrigine efficacy and tolerability as antidepressant augmentation for unipolar treatment-resistant depression compared to lithium. SUBJECTS AND METHODS: 88 patients suffering from treatment-resistant Major depressive disorder, having acute recurrent depressive episodes according to DSM IV criteria, were enrolled in the study. This was an open-label trial with a flexible dosing regimen. All patients, received antidepressants in full therapeutic doses. They were divided into two augmentation groups: 46 patients received 50-200 mg/day lamotrigine, and 42 patients received 600-1200 mg/day lithium. The Hamilton Rating Scale for Depression (HAM-D) and The Clinical Global Impression scale (CGI) were used to monitor therapeutic efficacy. Patients were evaluated weekly for an 8 week treatment period. RESULTS: The HAM-D total score was significantly reduced in both treatment groups at the study endpoint, without any difference between the groups. However, significant clinical improvement was reached within the second treatment week in the lamotrigine group compared to the lithium group (p=0.01 vs. lithium). Lamotrigine showed significant efficacy on the HAM-D item 1(depressed mood; p=0.01), item 7 (work and interest; p=0.01) and CGI-Improvement scale (p=0.02). The drop-out rate due to treatment failure was lower in the lamotrigine group (n=1) compared to the lithium (n=4) group. Also, the incidence of side effects did not differ between the groups. CONCLUSIONS: Our results suggest that lamotrigine could be useful as augmentation of antidepressants for treatment-resistant unipolar depression. Also, lamotrigine may accelerate the onset of antidepressant action, and therefore might be useful in treatment of major depression in general. PMID- 19556948 TI - Efficacy, tolerability and safety of tianeptine in special populations of depressive patients. AB - BACKGROUND: Tianeptine, a new generation antidepressant, possesses a unique mechanism of antidepressive action and has a specific pharmacokinetic profile. The aim of this study was to determine the efficacy, tolerability and safety of tianeptine in a "fragile" population of depressive patients: (1) a group of elderly patients and (2) a group with comorbid alcohol addiction. SUBJECTS AND METHODS: This was an open multicentric eight-week study of tianeptine efficacy, tolerability and safety including patients with mild to moderate depression (DSM IV), age > or =55 years (group 1; n=45) or with comorbid alcohol addiction (group 2; n=32). Assessments was made with the following rating scales; MADRS, HAM-A and CGI for efficacy and DESS for tolerability. RESULTS: After eight-week tianeptine therapy, remission (MADRS < or =12) was established in 51.1% and 84.4% patients, respectively. On day 7, the therapy led to a significant decrease of MADRS. On endpoint, there were significant differences on HAM-A, CGI-I and CGI-S scores (p<0.01). No adverse effects with frequency > or = 10%, were registered. A lower tolerability of tianeptine was registered in a group of elderly (nausea 4.5%, leg fatigue 4.4%, irritability 2.2%, bursts of crying and sadness 2.2%), while only 3.1% depressive patients with comorbid alcohol addiction had dizziness. CONCLUSION: This is the first clinical study to evaluate tolerability, efficacy and safety of tianeptine in a special population of depressive patients in the region. The study showed that tianeptine had good efficacy in treatment of mild to moderate forms of depression in special populations of depressive patients (elderly population and patients with comorbid alcohol addiction). The drug was well tolerated. PMID- 19556949 TI - The impact of stress related disorders on quality of life. AB - BACKGROUND: Some new studies have shown a significant connection between post traumatic stress disorder and impairment of the quality of life (QoL), whereas, to our knowledge, research on the connection between other disorders of the stress related group and quality of life do not exist. The objective of our study was to assess the connection between stress related disorders and the quality of life. SUBJECTS AND METHODS: We studied the quality of life in 160 subjects who have experienced certain stressogenic life events. Within the first group (80 subjects) a certain stress related disorder was diagnosed after a stressogenic life event. In the second group (80 subjects) none of the stress related disorders was developed after a stressogenic life event. The diagnosis was made based on the ICD-10 criteria; the stress was assessed by the Impact of Events Scale (IES) and QoL by the Manchester Short Assessment Quality of Life Scale (MANSA). RESULTS: Persons in whom some stress related disorders were diagnosed had a lower quality of life compared to persons who experienced stress but did not develop a disorder (p<0.01). CONCLUSION: Given the long-term stressogenic situation in our country, which is still ongoing, we think that comprehensive measures of prevention, early diagnostics and efficient treatment of stress related disorders are necessary in order to improve the quality of life for the persons with these disorders and their families, but also for society as a whole. PMID- 19556951 TI - Changing attitudes of high school students towards peers with mental health problems. AB - BACKGROUND: Stigma refers to the undesirable characteristics linked to mental illness and the adverse cognitive and behavioral consequences. Stigma causes a spiral of alienation and discrimination, leading to social isolation that diminishes chances for recovery. There is a great need for antistigma programs in order to decrease stigma related to persons with mental health problems. The antistigma program was initiated in schools of Serbia with the aim to address and decrease discrimination of adolescents with mental disorders. SUBJECTS AND METHODS: Sixty-three students from high schools voluntarily participated in the program. The effect of the program on the attitudes of students was evaluated by the Opinion about Mental Illness Questionnaire given to adolescents prior to its implementation and six months afterwards. RESULTS: Social discrimination and the tendency towards social restriction were reduced, while, at the same time, social awareness of mental health-related problems was increased among young people six months after program implementation. The results obtained clearly indicate positive changes in adolescents' attitudes and demonstrate a need for further educational activities regarding stigma and mental disorders. CONCLUSION: Stigma and discrimination reduction programs for adolescents are aimed at achieving a change of their attitudes toward the mental health problems of their peers and themselves through organized education. Our program demonstrates the necessity for youth participation in mental health services and system, and antistigma actions are seen as important aspects. PMID- 19556950 TI - How medical students in their pre-clinical year perceive psychiatry as a career: the study from Belgrade. AB - BACKGROUND: Taking the Initiative to evaluate students' affinity toward psychiatry seems to be a global issue and is an essential part of programs to improve the status of the profession. The aim of this study is to explore medical students' attitudes toward psychiatry in comparison to other residencies (internal medicine, surgery, pediatrics, gynecology and general medicine) in the pre-clinical year and to observe which factors influence the creation of these attitudes. SUBJECTS AND METHODS: The survey included 114 students of the second year, School of Medicine in Belgrade (academic year 2007/08). The data was collected trough a 23-item questionnaire. RESULTS: Fifteen percent of students stated that psychiatry was their career of choice, while 25% expressed a strong aversion. Psychiatry was ranked less attractive than internal medicine, surgery and pediatrics, but more attractive than general medicine or gynecology. Those who like psychiatry attributed more importance to an interesting and challenging job than to prestige and financial reward. Also, they found this field to be intellectually challenging and to rapidly expand the frontier of medicine. Students with negative attitude were convinced that psychiatry was lacking in scientific foundation and was clinically inefficient, they disliked intensive emotional involvement, exposure to stress and frequent unpleasant situations and had prejudices toward the patients or simply a lack of the interest. CONCLUSION: The present study is the first of its kind in Serbia which used a precise and internationally comparable methodological instrument and It shows that pre clinical medical students at the University of Belgrade, have a stronger affinity towards psychiatry when compared to their peers from most countries worldwide. Also, the study points out the fact that prejudices toward patients with mental dysfunctions and lack of confidence in the efficacy of psychiatric treatment should be specially targeted by the curriculum in the later part of undergraduate education. How this will affect the attitude of clinical students and graduates is to be examined. PMID- 19556952 TI - Sociodemographic profile of transsexual patients. AB - BACKGROUND: Gender identity disorder is a rare entity in psychiatry which affects deep instincts and personal identity. A review of the recent literature has shown that research of socio-demographic characteristics of transsexual patients is sporadic. There are very few investigations which consider the socio-demographic characteristics in individuals with disharmonized sex and gender identity. This kind of research has not been done in Serbia until now. The objective of this research was to evaluate and analyze socio-demographic data of transsexual persons with a homosexual orientation. SUBJECT AND METHODS: The paper presents the mentioned characteristics on sample of 30 gender dysphoric persons. The patients were examined by The Belgrade team for gender identity. The transsexuals were in a stage of preparation for the sex-reassignment surgery (SRS), and transsexualism was diagnosed and guided within this program (used criteria were keeping with ICD X, DSM IV and Diagnostic schedule recommended by Standards of Care for Dysphoric Persons of the Harry Benjamin International Gender Dysphoria Association). We have examined the following socio-demographic characteristics: sex, age, place of residence, educational level, employment and religion. RESULTS: Results and analysis of obtained data have shown that the socio demographic profile of transsexual patients included in the research is: predominantly males (male/female sex ratio 3:2); refer for psychiatric help in younger age (frequently before 26); most of them with high school education; live in urban communities; mostly Orthodox religion; equally employed and unemployed. CONCLUSION: Gathering and analysis of sociodemographic data is important for elucidating the transsexual patient's profile. It facilitates better understanding, timely recognition and choice of appropriate treatment for these patients. PMID- 19556953 TI - Use or underuse of therapeutic guidelines in psychiatry? AB - A rapid expansion of new treatment options in recent decades and the increasing volume of scientific evidence published on a daily basis have been followed by the necessity of introduction of clinical guidelines and therapeutic algorithms. The development of these guidelines and algorithms has also been driven by increased cost-awareness and the increasing pressure to improve cost-efficiency. The Serbian Physicians Society published "Therapeutic Guidelines for the Treatment of Schizophrenia" in 2003 and "Therapeutic Guidelines for the Treatment of Bipolar Affective Disorder" in 2004. The School of Medicine, University of Belgrade published "Therapeutic Guidelines for the Treatment of Depression" in 2004. All of these national guidelines, at the moment of development, were based upon up-to-date scientific evidence. According to the recently conducted survey at the Institute of Psychiatry, Clinical Centre of Serbia, about 65% of psychiatrists stated that they adhere to the national or relevant international therapeutic guidelines. When asked to cite which international or foreign guidelines in particular they used, approximately 50% failed to do so, while the other half cited mostly the APA Guidelines or NICE Guidelines. Among the national guidelines, physicians are, according to the survey, familiar with the Therapeutic Guidelines for the treatment of Schizophrenia (46.3%), Therapeutic Guidelines of Depression (41.5%) and Therapeutic Guidelines for the Treatment of Bipolar Affective Disorder (34.1%). The majority of Serbian psychiatrists rely on the efficacy and safety of the drugs as the major determining factors in the choice of therapy, bearing in mind the patients' best interests. However, it is unclear why such a discrepancy between practice and guidelines still persists, since guidelines also recommend therapy based on their safety and efficacy data. It is possible that a substantial percentage of psychiatrists obtain indicators on drugs' efficacy and safety from their personal professional experience. It is doubtful whether this knowledge is valid, or just represents unproven prescribers' habits. Furthermore, the influence on other factors, such as treatment costs or drug availability should be further investigated. PMID- 19556954 TI - Psychiatry and movies. AB - As one of the most potent and substantial form of mass communication, film exercises a very significant influence upon the perceptions of the audience, especially in relation to mental illness issues, and that perception is very much blurred with populists' misinterpretation and lack of awareness regarding problems faced by persons suffering from mental disorders. Movies such as "Psycho", "One Flew Over Cuckoo's Nest", "Exorcist", despite being valuable in an artistic sense, corroborated and encouraged confusion and undermined the clarity and certainty concerning the fine line separating mental health from mental illness. Modern film makers and movie theoreticians try to overcome these limitations which are often generated by exploitation of stereotypes and myths referring to mentally ill people. This paper defines and discusses the most frequent thematic stereotypes seen in movies which are perpetuating stigmatization of mentally ill people. They are: free-spirited rebel, maniac on a killing spree, seducer, enlightened member of society, narcissistic parasite, beastly person (stereotype of animal sort). Psychiatry and cinematography are linked inseparably not only because they creatively complement each other, but also as an opportunity of mutual influences blending into didactical categories and professional driving forces, benefiting both the filmmakers' and the psychiatrists' professions. PMID- 19556955 TI - Animal-assisted therapy--a new trend in the treatment of children and adults. AB - Animal-assisted therapy is a familiar method of treatment in the rehabilitation of many illnesses and conditions, but is still not applied sufficiently in our milieu. This paper gives an overview of the available literature and some of the research which demonstrates that the interaction between the patient, animal and therapist provides a context which improves communication, elevates self confidence, reduces the symptoms of diseases, and improves the quality of life. The dog, cat, horse, birds and toy animals are most often used in therapy. Short term contacts with animals are used, as well as long term keeping of animals, which are looked after by patients following a particular methodology. The therapy is used in the treatment of psychiatric patients afflicted with depression, schizophrenia, phobias and addiction problems. Loneliness is easier to endure in the company of animals. It is also applied in cardiovascular diseases, dementia, Alzheimer's disease, child cerebral paralysis, rheumatoid arthritis, AIDS, and other diseases. Research shows a more rapid reduction of symptoms of many diseases when animals are included in the therapeutic process. PMID- 19556956 TI - Case report of autogynophilia--family, ethical and surgical implications. AB - Through the case presentation of a diagnostically and therapeutically interesting gender dysphoric individual, the authors wish to address diagnostic problems associated with this controversial category, illustrate dilemmas and emphasize the importance of diagnostic procedures in differentiating between primary transsexualism and other transgender states. Many questions have been triggered by this case, mainly about whether this patient should be classified as a paraphilia (transvestite, transvestite with transsexual trend), primary transsexualism or autogynephilia and about the most adequate treatment (e.g., sex reassignment surgery, hormone therapy as a way of partial feminisation or exclusively psychotherapy). The issue of reconstructive surgery, i.e. its justification in the case of this particular condition is specifically discussed. Before any decision is made, both medical but also ethical consequences of the treatment choice need to be considered (e.g., the client is the father of two underage children). PMID- 19556959 TI - Melanoma: promising new discoveries and treatment modalities for difficult clinical scenarios - part II. PMID- 19556957 TI - Accidental death due to complete autoerotic asphyxia associated with transvestic fetishism and anal self-stimulation - case report. AB - A case is reported of a 36-year-old male, found dead in his locked room, lying on a bed, dressed in his mother's clothes, with a plastic bag over his head, hands tied and with a barrel wooden cork in his rectum. Two pornographic magazines were found on a chair near the bed, so that the deceased could see them well. Asphyxia was controlled with a complex apparatus which consisted of two elastic luggage rack straps, the first surrounding his waist, perineum, and buttocks, and the second the back of his body, and neck. According to the psychological autopsy based on a structured interview (SCID-I, SCID-II) with his father, the deceased was single, unemployed and with a part college education. He had grown up in a poor family with a reserved father and dominant mother, and was indicative of fulfilling DSM-IV diagnostic criteria for alcohol dependence, paraphilia involving hypoxyphilia with transvestic fetishism and anal masturbation and a borderline personality disorder. There was no evidence of previous psychiatric treatment. The Circumstances subscale of Beck's Suicidal Intent Scale (SIS-CS) pointed at the lack of final acts (thoughts or plans) in anticipation of death, and absence of a suicide note or overt communication of suicidal intent before death. Integration of the crime scene data with those of the forensic medicine and psychological autopsy enabled identification of the event as an accidental death, caused by neck strangulation, suffocation by a plastic bag, and vagal stimulation due to a foreign body in the rectum. PMID- 19556960 TI - Epigenetics in human melanoma. AB - BACKGROUND: Recent technological advances have allowed us to examine the human genome in greater detail than ever before. This has opened the door to an improved understanding of the gene expression patterns involved with cancer. METHODS: A review of the literature was performed to determine the role of epigenetic modifications in human melanoma. We focused the search on histone deacetylation, methylation of gene promoter regions, demethylation of CpG islands, and the role of microRNA. We examined the relationship between human melanoma epigenetics and their importance in tumorigenesis, tumor progression, and inhibition of metastasis. The development and clinical application of select pharmacologic agents are also discussed. RESULTS: We identified several articles that have extensively studied the role of epigenetics in melanoma, further elucidating the complex processes involved in gene regulation and expression. Several new agents directly affect epigenetic mechanisms in melanoma, with divergent affects on the metastatic potential of melanoma. CONCLUSIONS: Epigenetic mechanisms have emerged as having a central role in gene regulation of human melanoma, including the identification of several putative tumor suppressor genes and oncogenes. Further research will focus on the development of novel therapeutics that will likely target and alter such epigenetic changes. PMID- 19556961 TI - Tumor biomarkers in melanoma. AB - BACKGROUND: Morphologic and histopathologic markers have been the backbone for the classification and prognostic assessment of melanoma. Availability of an increasing number of molecular markers, however, provides the potential for refining diagnostic and prognostic categories in this disease. METHODS: We reviewed the recent data that are accumulating concerning gene expression and genetic profiling and related these to clinical aspects of the disease. RESULTS: Multiple biomarkers have now been described, and their biologic significance is being established. In addition, several candidate molecules involved in melanoma pathogenesis have been identified. CONCLUSIONS: The process of biomarker identification and validation is providing a rapidly changing molecular view of melanoma, a strategy that is necessary for developing truly stratified or even personalized prevention or management. PMID- 19556962 TI - Pediatric melanoma: a review. AB - BACKGROUND: Malignant melanoma is a rare neoplasm in the pediatric population, but its incidence has risen in recent years. METHODS: The literature was reviewed to define the current clinical and pathologic features of pediatric melanoma, highlighting the similarities and differences between adult and pediatric melanoma. RESULTS: Distinctive features of this disease, including frequency and type of genetic abnormalities, predisposing conditions, clinical presentation, stage at diagnosis, prognostic features, and frequency of sentinel lymph node positivity are emphasized. Treatment strategies, extrapolated from adult melanoma trials, are also discussed. CONCLUSIONS: Despite the differences between pediatric and adult melanoma, survival rates are similar and are improving in both populations. Further studies will help delineate the pathogenesis of both adult and pediatric melanoma, with the goal of contributing to early detection and improved survival. PMID- 19556963 TI - Sentinel lymph node biopsy for melanoma: indications and rationale. AB - BACKGROUND: The disease status of regional lymph nodes is the most important prognostic indicator for patients with melanoma. Sentinel lymph node biopsy (SLNB) was developed as a technique to surgically assess the regional lymph nodes and spare node-negative patients unnecessary and potentially morbid complete lymphadenectomies. METHODS: We reviewed the literature on SLNB for cutaneous melanoma to provide insight into the rationale for the current widespread use of SLNB. RESULTS: Multiple studies show that the status of the SLN is an important prognostic indicator. Those with positive SLNs have significantly decreased disease-free and melanoma-specific survival compared with those who have negative SLNs. In the Multicenter Selective Lymphadenectomy Trial I (MSLT-I), in which patients with intermediate-thickness melanoma were randomized to SLNB (and immediate completion lymphadenectomy if the SLN was positive) vs observation (and a lymphadenectomy only after presenting with clinically evident recurrence), the 5-year survival rate was 72.3% for patients with positive sentinel nodes and 90.2% for those with negative sentinel nodes (P < .001). Although overall survival was not increased in patients who underwent SLNB compared with those who were randomized to observation, patients who underwent SLNB had a significantly increased 5-year disease-free survival rate compared with those who underwent observation alone (78.3% in the biopsy group and 73.1% in the observation group; P = .009). For those with nodal metastases, patients who underwent SLNB and immediate lymphadenectomy had an increased overall 5-year survival rate compared with those who had lymphadenectomy only after presenting with clinically evident disease (72.3% vs 52.4%; P = .004). Moreover, other studies show that for patients with thin melanomas or= 0.76 mm and increased mitotic rate have been shown to be associated with an increased incidence of SLN metastases. CONCLUSIONS: SLNB provides important prognostic and staging data with minimal morbidity and can be used to identify regional node-negative patients who would not benefit from a complete nodal dissection. In our opinion, SLNB should be performed on most patients (with acceptable surgical and anesthesia risk) who have melanomas with a Breslow depth >or= 0.76 mm. PMID- 19556964 TI - Limiting the morbidity of inguinal lymphadenectomy for metastatic melanoma. AB - BACKGROUND: Surgery is currently the primary treatment modality for metastatic melanoma involving the inguinal lymph nodes. However, inguinal lymph node dissections are associated with substantial morbidity including infection, wound dehiscence, lymphedema, seroma, and deep venous thromboembolism (DVT). Improved understanding is needed regarding the factors predisposing patients to complications and the operative and perioperative maneuvers that can decrease morbidity. METHODS: We reviewed recently published literature regarding the morbidity associated with lymphadenectomy in the treatment of inguinal metastatic melanoma. Where available, emphasis was focused on appropriately designed studies aimed at reducing treatment-related morbidity. When appropriate, the review was supplemented by our personal experience. RESULTS: Strategies to limit treatment related morbidity involve optimizing the preoperative assessment, operative technique, and postoperative care. Establishing the diagnosis of nodal metastasis early using minimally invasive techniques is critical to reduce subsequent perioperative complications. Morbidity is higher for inguinal compared to cervical or axillary lymphadenectomy, and many variations in extent of inguinal lymphadenectomy and operative technique have been reported. The lack of definitive trials has led to controversy regarding surgical technique such as indications for pelvic lymphadenectomy ("deep" node dissection), saphenous vein preservation, and sartorius transposition. In the postoperative period, the use of DVT and lymphedema prophylaxis should be considered to potentially improve patient outcomes. CONCLUSIONS: While the morbidity of inguinal lymphadenectomy can be substantial, several straightforward pre- and postoperative measures can be instituted to limit morbidity. Controversy persists regarding the indications for and benefit of pelvic lymphadenectomy, saphenous vein preservation, and sartorius muscle transposition. A multi-institutional trial is currently in progress to investigate the safety of avoiding lymphadenectomy in patients with microscopic metastases in the sentinel node. PMID- 19556965 TI - Diagnosis and treatment of melanoma brain metastasis: a literature review. AB - BACKGROUND: Brain metastasis is common in patients with malignant melanoma and represents a significant cause of morbidity and mortality. Nearly 37% of patients with malignant melanoma eventually develop brain metastasis, and autopsy reports show that 75% of those who died of this disease developed brain metastasis. METHODS: We review the level I and level II evidence that guides indications for treatment with surgery, stereotactic radiosurgery, chemotherapy, and immunotherapy for patients with melanoma brain metastasis. RESULTS: Level I evidence supports the role of whole brain radiotherapy, microsurgery, and radiosurgery alone or in combination for the treatment of patients with melanoma brain metastasis. Chemotherapy has been ineffective. Ongoing studies continue to assess the effects of immunotherapy and agents in development. CONCLUSIONS: Brain metastasis is a common and formidable challenge in patients with malignant melanoma. Although there have been no randomized controlled trials exclusively in patients with melanoma brain metastasis, care can be guided by the application of level I evidence for the treatment of brain metastasis in general and phase II studies focusing specifically on melanoma brain metastasis. Promising new agents and approaches are needed and will hopefully be identified in the near future. PMID- 19556966 TI - The use of sociocultural constructs in cancer screening research among African Americans. AB - BACKGROUND: Studies are increasingly examining the role of sociocultural values, beliefs, and attitudes in cancer prevention. However, these studies vary widely in how sociocultural constructs are defined and measured, how they are conceived as affecting cancer beliefs, behaviors, and screening, and how they are applied in interventions. METHODS: To characterize the current state of this research literature, we conducted a critical review of studies published between 1990 and 2006 to describe the current use of sociocultural constructs in cancer screening research among African Americans. We included quantitative and qualitative studies with cancer as a primary focus that included African American participants, assessed screening behaviors, reported race-specific analyses, and considered one or more sociocultural factors. Studies were evaluated for type of cancer and screening analyzed, study population, methodology, sociocultural constructs considered, definitions of constructs, provision of psychometric data for measures, and journal characteristics. RESULTS: Of 94 studies identified for review, 35 met the inclusion criteria and were evaluated. Most focused on breast cancer screening, and thus African American women. Sociocultural constructs were seldom clearly defined, and the sources and psychometric properties of sociocultural measures were rarely reported. CONCLUSIONS: A multidisciplinary approach to developing a common language and a standardized set of measures for sociocultural constructs will advance research in this area. Specific recommendations are made for future research. PMID- 19556967 TI - Measuring replicative life span in the budding yeast. AB - Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The budding yeast Saccharomyces cerevisiae has been used extensively to study the biology of aging, and several determinants of yeast longevity have been shown to be conserved in multicellular eukaryotes, including worms, flies, and mice. Due to the lack of easily quantified age-associated phenotypes, aging in yeast has been assayed almost exclusively by measuring the life span of cells in different contexts, with two different life span paradigms in common usage. Chronological life span refers to the length of time that a mother cell can survive in a non-dividing, quiescence-like state, and is proposed to serve as a model for aging of post mitotic cells in multicellular eukaryotes. Replicative life span, in contrast, refers the number of daughter cells produced by a mother cell prior to senescence, and is thought to provide a model of aging in mitotically active cells. Here we present a generalized protocol for measuring the replicative life span of budding yeast mother cells. The goal of the replicative life span assay is to determine how many times each mother cell buds. The mother and daughter cells can be easily differentiated by an experienced researcher using a standard light microscope (total magnification 160X), such as the Zeiss Axioscope 40 or another comparable model. Physical separation of daughter cells from mother cells is achieved using a manual micromanipulator equipped with a fiber-optic needle. Typical laboratory yeast strains produce 20-30 daughter cells per mother and one life span experiment requires 2-3 weeks. PMID- 19556968 TI - Far-red light inhibits germination through DELLA-dependent stimulation of ABA synthesis and ABI3 activity. AB - Under the canopy, far-red (FR) light represses seed germination by inactivating phytochrome photoreceptors. This elicits a decrease in gibberellins (GA) levels and an increase in abscisic acid (ABA) levels. GA promotes germination by enhancing the proteasome-mediated destruction of DELLA repressors. ABA prevents germination by stimulating the expression of ABI repressors. How phytochromes elicit changes in hormone levels or how GA- and ABA-dependent signals are coordinated to repress germination remains poorly understood. We show that repression of germination by FR light involves stabilized DELLA factors GAI, RGA and RGL2 that stimulate endogenous ABA synthesis. In turn, ABA blocks germination through the transcription factor ABI3. The role of PIL5, a basic helix-loop-helix transcription factor stimulating GAI and RGA expression, is significant, provided GA synthesis is high enough; otherwise, high GAI and RGA protein levels persist to block germination. Under white light, GAI and RGA driven by the RGL2 promoter can substitute for RGL2 to promote ABA synthesis and repress germination, consistent with the recent findings with RGL2. The three DELLA factors inhibit testa rupture whereas ABI3 blocks endosperm rupture. PMID- 19556969 TI - Unusual bipartite mode of interaction between the nonsense-mediated decay factors, UPF1 and UPF2. AB - Nonsense-mediated decay (NMD) is a eukaryotic quality control mechanism that degrades mRNAs carrying premature stop codons. In mammalian cells, NMD is triggered when UPF2 bound to UPF3 on a downstream exon junction complex interacts with UPF1 bound to a stalled ribosome. We report structural studies on the interaction between the C-terminal region of UPF2 and intact UPF1. Crystal structures, confirmed by EM and SAXS, show that the UPF1 CH-domain is docked onto its helicase domain in a fixed configuration. The C-terminal region of UPF2 is natively unfolded but binds through separated alpha-helical and beta-hairpin elements to the UPF1 CH-domain. The alpha-helical region binds sixfold more weakly than the beta-hairpin, whereas the combined elements bind 80-fold more tightly. Cellular assays show that NMD is severely affected by mutations disrupting the beta-hairpin binding, but not by those only affecting alpha-helix binding. We propose that the bipartite mode of UPF2 binding to UPF1 brings the ribosome and the EJC in close proximity by forming a tight complex after an initial weak encounter with either element. PMID- 19556971 TI - Prior exercise lowers blood pressure during simulated night-work with different meal schedules. AB - BACKGROUND: Shift-work and a sedentary lifestyle are risk factors for raised blood pressure (BP). Exercise can reduce BP in diurnally-active individuals, but it is unknown whether postexercise hypotension persists when people are active and eating at night. We present the first investigation into the acute effects of exercise on BP monitored during simulated night-work. METHODS: Nine normotensive participants, aged 20-42 years, completed at least two crossover trials beginning at 1800 hours. Between 1900 and 2000 hours, participants either rested or exercised at 50% peak oxygen uptake (VO(2peak)) and then remained awake throughout the night, completing various tasks until 0515 hours. Six participants completed a total of four trials in which they exercised or rested, whereas either one standardized (60 kJ/kg) meal at 2200 hours or two smaller (30 kJ/kg) meals at 2200 and 0200 hours were eaten. Systolic and diastolic BP, mean arterial pressure (MAP), heart rate (HR), and wrist activity were recorded every 30 min. RESULTS: Following exercise, MAP was significantly (P < 0.0005) lower throughout the night-shift compared with no prior exercise (95% confidence limits for reduction: 4-7 mm Hg). The postexercise reductions in systolic BP and MAP were not moderated by diet, but the reduction in diastolic BP was slightly greater when only one meal was eaten (P < 0.0005). BP was lower even though wrist activity and HR were significantly higher following exercise (P < 0.0005). CONCLUSIONS: These data indicate that prior exercise lowers BP throughout a subsequent 8-h night-shift in healthy individuals within the normotensive range. Therefore, regular low-intensity exercise might moderate the well-known association between shift-work participation and raised BP. PMID- 19556970 TI - Structure and function of a complex between chorismate mutase and DAHP synthase: efficiency boost for the junior partner. AB - Chorismate mutase catalyzes a key step in the shikimate biosynthetic pathway towards phenylalanine and tyrosine. Curiously, the intracellular chorismate mutase of Mycobacterium tuberculosis (MtCM; Rv0948c) has poor activity and lacks prominent active-site residues. However, its catalytic efficiency increases >100 fold on addition of DAHP synthase (MtDS; Rv2178c), another shikimate-pathway enzyme. The 2.35 A crystal structure of the MtCM-MtDS complex bound to a transition-state analogue shows a central core formed by four MtDS subunits sandwiched between two MtCM dimers. Structural comparisons imply catalytic activation to be a consequence of the repositioning of MtCM active-site residues on binding to MtDS. The mutagenesis of the C-terminal extrusion of MtCM establishes conserved residues as part of the activation machinery. The chorismate-mutase activity of the complex, but not of MtCM alone, is inhibited synergistically by phenylalanine and tyrosine. The complex formation thus endows the shikimate pathway of M. tuberculosis with an important regulatory feature. Experimental evidence suggests that such non-covalent enzyme complexes comprising an AroQ(delta) subclass chorismate mutase like MtCM are abundant in the bacterial order Actinomycetales. PMID- 19556972 TI - Aliskiren monotherapy results in the greatest and the least blood pressure lowering in patients with high- and low-baseline PRA levels, respectively. AB - Hypertensive patients with low-baseline plasma renin activity (PRA) are known to respond best to natriuretic drugs, and those with high PRA respond best to renin angiotensin system (RAS) blockade. However, there has been recent speculation that blood pressure (BP)-lowering responses to the renin inhibitor, aliskiren, might also be blunted in some patients with medium-to-high baseline PRA. It has been suggested that treatment resistance in these patients may result from excessive reactive increases in renin secretion, such that aliskiren's blockade of PRA is overwhelmed. In order to test for evidence in support of this hypothesis, we conducted a reanalysis of original data from three published clinical trials of aliskiren. When aliskiren was administered as a monotherapy, or in combination with other blockers of the RAS, changes in PRA were closely correlated with baseline PRA. Patients with low-baseline PRA demonstrated small reductions or rises in PRA, rather than patients with medium-to-high baseline PRA. We confirmed that ambulatory BP-lowering responses to full dose aliskiren monotherapy were greatest and least among patients with high- and low-baseline PRA, respectively. However no such association was demonstrated during aliskiren combination therapy. With either monotherapy or combination therapy, no patient with a baseline PRA >0.65 ng/ml/h was observed to have a rise in both PRA and BP. We conclude, therefore, that there is only evidence for one type of resistance to aliskiren--as with all blockers of the RAS, lesser BP-lowering responses to aliskiren occur in those with the least renin to block. PMID- 19556973 TI - Association of highly active antiretroviral therapy with increased arterial stiffness in patients infected with human immunodeficiency virus. AB - BACKGROUND: Metabolic disorders associated with atherosclerosis and cardiovascular disease have been described in "HIV-infected" individuals. We investigated (i) whether normotensive "HIV-infected" individuals and hypertensive patients have similarities regarding their arterial elastic properties and (ii) the effect of highly active antiretroviral therapy (HAART) and metabolic factors on arterial stiffness. METHODS: In a case-control study, we compared measurements of pulse wave velocity (PWV), arterial blood pressure, and markers of metabolic profile in 56 normotensive, "HIV-infected" patients (mean age 40 +/- 13 years) to 28 age- and sex-matched newly diagnosed untreated patients with hypertension and 28 healthy individuals. RESULTS: "HIV-infected" patients had higher PWV than healthy controls but lower PWV than hypertensives (8.1 +/- 1.4 m/s vs. 6.7 +/- 1.1 m/s vs. 9.0 +/- 1.0 m/s, P = 0.003 and 0.01, respectively). However, patients on HAART had similar PWV with hypertensives (8.4 +/- 1.4 vs. 9.0 +/- 1.0 m/s P = 0.25). Patients on HAART had higher PWV than patients without (8.4 +/- 1.4 m/s vs. 7.5 +/- 1.3 m/s, P = 0.03). Patients on HAART had higher total cholesterol, triglycerides, and diastolic blood pressure than patients naive to HAART (P < 0.05). In multivariate analysis, the independent determinants of increased PWV were HAART duration (unstandardized coefficient b v = 0.007, P = 0.04), serum cholesterol (b = 0.007, P = 0.04), mean or diastolic blood pressure (b = 0.049 and b = 0.060, P < 0.01). CONCLUSIONS: "HIV-infected" individuals have increased arterial stiffness compared to healthy controls. Patients on antiretroviral therapy have similarities regarding their arterial elastic properties with patients with untreated hypertension. There is an independent association between duration of antiretroviral therapy, cholesterol levels, and blood pressure with increased arterial stiffness in "HIV-infected" patients. PMID- 19556974 TI - Retinol-binding protein-4 in women with untreated essential hypertension. AB - BACKGROUND: Retinol-binding protein-4 (RBP4) is a novel adipokine able to modulate the action of insulin in several tissues. A variable degree of insulin resistance characterizes the vast majority of hypertensive (HYP) patients. The aim of this study was to evaluate the relationship between RBP4 and essential hypertension, exploring potential links between RBP4 and other adipokines with some proxies of early vascular damage in female naive HYP patients. METHODS: Serum RBP4, leptin, adiponectin, and resistin levels were determined in 35 HYP and 35 normotensive lean women with normal glucose tolerance paired by age and body mass index (BMI) served as controls (CTL); carotid intima-media thickness (IMT) was also measured. RESULTS: A striking difference was observed in RBP4 levels between HYP and CTL with significantly higher levels in the former than in the latter. No relationship was observed between glomerular filtration rate (GFR) and RBP4. Adiponectin levels were slightly but significantly lower in HYP than in CTL, whereas no differences were observed in resistin and leptin concentrations between the two groups of women. In the whole study group, a strong linear relationship was observed between IMT value and both RBP4 (rho = 0.321, P = 0.0076) and resistin (rho = 0.340, P = 0.0048); these two adipocytokines, together with cholesterol, were the only variables independently related to IMT (r(2) = 0.24; P = 0.004) by a stepwise analysis. CONCLUSIONS: RBP4 levels are increased in naive HYP women and correlated with the degree of IMT suggesting a participation of this adipocytokine in the modulation of the atherosclerotic process exerted by the adipose tissue as endocrine organ. PMID- 19556975 TI - Climate burden of refrigerants rockets. PMID- 19556977 TI - The bias in self-reported obesity from 1976 to 2005: a Canada-US comparison. AB - The objective of this study was to determine whether the bias in self-reported estimates of obesity has changed over time and followed different patterns in Canada and the United States. Using age-standardized data from three waves of the National Health and Nutrition Examination Survey (NHANES) in the United States and the Canadian Community Health Survey (CCHS) and the Canadian Heart Health Survey (CHHS) in Canada, discrepancies were compared between reported and measured estimates of height, weight, and obesity (based on the BMI) from 1976 to 2005. Results indicated that obesity increased in both countries, but rates were higher in the United States. The discrepancy between self-reported and measured obesity was small in the United States with reported data underestimating measured prevalence by about 3%; this stayed relatively constant over time. In Canada, the discrepancy was large and doubled in the past decade (from 4 to 8%). In the United States, self-reported data may be more accurate in monitoring changes in obesity over time, as the estimates have consistently remained about 3% below the measured estimates, whereas in Canada, monitoring obesity based solely on self-reported height and weight may produce inaccurate estimates because of the increasing discrepancy between self-reported and measured data. PMID- 19556976 TI - Roux-en-Y gastric bypass enhances energy expenditure and extends lifespan in diet induced obese rats. AB - Gastrointestinal weight-loss surgery (GIWLS) is currently the most effective treatment for severe obesity, with Roux en-Y gastric bypass (RYGB) among the best of the available surgical options. Despite its widespread clinical use, the mechanisms by which RYGB induces its profound weight loss remain largely unknown. This procedure effects weight loss by altering the physiology of weight regulation and eating behavior rather than by simple mechanical restriction and/or malabsorption as previously thought. To study how RYGB affects the physiology of energy balance, we developed a rat model of this procedure. In this report, we demonstrate that RYGB in diet-induced obese (DIO) rats induces a 25% weight loss, prolongs mean survival by 45%, and normalizes glucose homeostasis and lipid metabolism. RYGB induced a 19% increase in total and a 31% increase in resting energy expenditure (REE). These effects, along with a 17% decrease in food intake and a 4% decrease in nutrient absorption account for the normalization of body weight after this procedure. These effects indicate that surgery acts by altering the physiology of weight regulation and help to explain the effectiveness of RYGB in comparison to restrictive dieting and other forms of dietary and pharmacological therapies for obesity. The clinical effectiveness of RYGB and its physiological effects on body weight regulation and energy expenditure (EE) suggest that this operation provides a unique opportunity to explore the mechanisms of energy homeostasis and to identify novel therapies for obesity and related metabolic diseases. PMID- 19556978 TI - Proteomic analysis of human adipose tissue after rosiglitazone treatment shows coordinated changes to promote glucose uptake. AB - The aim of this study was to identify potential protein targets for insulin sensitization in human adipose tissue using unbiased proteomic approaches. Ten moderately obese, but otherwise healthy, subjects were treated with rosiglitazone 4 mg b.i.d. for 14 days and global protein and gene expression changes were monitored. Proteomic analysis revealed distinct up- or downregulation (greater than twofold) in 187 protein spots on the two-dimensional (2-D) gel images between day 0 and day 1 adipose tissue samples. When comparing the protein spots on the gels from day 0 with that of 14-day-treated samples, 122 spots showed differential expression. There was a striking increase in the expression of proteins involved in glucose transporter-4 (GLUT4) granule transport and fusion (actin, myosin-9, tubulin, vimentin, annexins, moesin, LIM, and SH3 domain protein-1), signaling (calmodulin, guanine nucleotide-binding proteins), redox regulation (superoxide dismutase, catalase, ferritin, transferrin, heat shock proteins), and adipogenesis (collagens, galectin-1, nidogen-1, laminin, lamin A/C). However, there was an intriguing absence of correlated changes in mRNA expression, suggesting adaptation at a post-transcriptional level in response to rosiglitazone. Thus, the major changes observed were among proteins involved in cytoskeletal rearrangement, insulin and calcium signaling, and inflammatory and redox signals that decisively upregulate GLUT4 granule trafficking in human adipose tissue. Such orchestrated changes in expression of multiple proteins provide insights into the mechanism underlying the increased efficiency in glucose uptake and improvement of insulin sensitivity in response to rosiglitazone treatment. PMID- 19556979 TI - Affective responses to increasing levels of exercise intensity in normal-weight, overweight, and obese middle-aged women. AB - At least 60 min of daily physical activity (PA) are recommended for weight control, a target achieved by only 3% of obese (OB) women. The purposes of this study were to examine (i) the affective responses of normal-weight (NW), overweight (OW), and OB middle-aged sedentary women to exercise of increasing intensity and (ii) the relationship of affective responses to self-efficacy and social physique anxiety. The women participated in a graded treadmill protocol to volitional exhaustion while providing ratings of pleasure-displeasure and perceived activation each minute. The Activation Deactivation Adjective Check List (AD ACL) was also completed before and after exercise. The affective responses of NW and OW women did not differ. However OB women gave lower pleasure ratings during the incremental protocol and reported lower Energy scores immediately after the protocol. Social physique anxiety, but not self-efficacy, was inversely related to pleasure and energy. The lower levels of pleasure and energy experienced by OB than nonobese women could account in part for their dramatically low levels of PA participation. Modifying the cognitive antecedents of social physique anxiety might be a useful intervention strategy. PMID- 19556980 TI - A cell biological view of Toll-like receptor function: regulation through compartmentalization. AB - An emerging paradigm in innate immune signalling is that cell biological context can influence the outcome of a ligand-receptor interaction. In this Review we discuss how Toll-like receptor (TLR) activation and signal transduction are regulated by subcellular compartmentalization of receptors and downstream signalling components. In particular, we focus on the functional specialization of TLRs in the endosomal system. We discuss recent studies that illustrate how basic aspects of the cellular machinery contribute to TLR function and regulation. This emerging area of research will provide important information on how immune signal transduction networks depend on (and in some cases influence) the generic regulators that organize eukaryotic cells. PMID- 19556982 TI - Selective resuscitation in premature twins: an ethical analysis. AB - Selective resuscitation refers to the practice of providing resuscitative efforts to one or some (but not all) infants born in the setting of multiple gestation. When one fetus is known to have a severe anomaly or severe growth restriction, parents are sometimes offered this option. In the setting of extreme prematurity, in the absence of an anomaly or severe growth restriction, parents are generally expected to make one unified decision for all the infants involved. The introduction of the Outcome Estimator, a tool that provides the ability to make individual outcome predictions for each fetus in a multiple gestation at borderline gestational age, based on contributing variables such as weight and gender, has led to the ethical dilemma of whether parents in this setting should also be offered the option of selective resuscitation. No convincing ethical argument for denying the parents the right to decide for each individual infant is apparent. PMID- 19556981 TI - Optimizing protein intake in preterm infants. PMID- 19556983 TI - The fetal heart monitor tracing in pregnancies complicated by a spontaneous umbilical cord hematoma. AB - The objective of this study is to present and describe the fetal heart rate appearance in pregnancies complicated by an antenatal spontaneous umbilical cord hematoma that resulted in a live birth. Three cases of antenatal spontaneous umbilical cord hematoma are described. All three patients presented with a complaint of decreased fetal movement. The fetal heart monitor tracings on admission are depicted and discussed. In all three cases, the fetal heart rate pattern showed decreased variability with an absence of accelerations. Decelerations were noted but were identified in 25% or less of the contractions and primarily with contractions that exceeded 90 s. Absent accelerations with minimal to absent variability, if caused by uteroplacental insufficiency, usually develop in the presence of recurrent decelerations. Absent accelerations with minimal to absent variability in the absence of recurrent decelerations may suggest other causes including aneuploidy or congenital cardiac or neurologic anomalies. Though rare, spontaneous umbilical cord hematoma can be added to the differential. PMID- 19556984 TI - Intrauterine subdural hemorrhage in a preterm neonate possibly associated with maternal low-molecular weight heparin treatment. AB - We report intrauterine subdural hemorrhage in a preterm infant delivered by cesarean section at 32 weeks following vaginal bleeding of a mother treated with low-molecular weight heparin (LMWH) for deep vein thrombosis. The subdural hematomas were partially calcified, proving antenatal occurrence. Maternal trauma during pregnancy, intrauterine infection, cerebral vascular malformation and congenital coagulopathy as known etiologies of subdural hemorrhage could be ruled out. Intrauterine subdural hemorrhage may be an exceptional complication of maternal LMWH treatment. PMID- 19556985 TI - Extralobar sequestration. PMID- 19556987 TI - Secondary adrenal insufficiency in sick neonates: should we treat with ACTH? PMID- 19556989 TI - The fallacy of 'alternative' medicine. PMID- 19556990 TI - Proteinuria: Increased angiotensin-receptor blocking is not the first option. AB - Candesartan doses in excess of the recommended antihypertensive maxima have been reported to lead to greater reductions of proteinuria than the advised doses. High-dose angiotensin-converting-enzyme inhibitors, however, are at least as effective as high-dose angiotensin blockers and less expensive. Angiotensin converting-enzyme inhibition is thus the first-line strategy to halt kidney disease progression. PMID- 19556992 TI - Diabetes: Renal benefits of lowering BP in the absence of hypertension. AB - Regardless of baseline blood pressure, treatment with a combination of an angiotensin-converting enzyme inhibitor and a diuretic decreases incidence of renal events in patients with type 2 diabetes. The combination therapy approach could be the key to achieving renoprotection. PMID- 19556991 TI - Dialysis: Phosphorus binders and survival: need for randomized trials. AB - An observational study suggests that administration of phosphorus binders dramatically improves survival rates in patients on incident hemodialysis-even in those without hyperphosphatemia. Randomized clinical trials should drive changes in the relevant clinical practice. PMID- 19556993 TI - Transplantation: To convert or not to convert: lessons from the CONVERT trial. AB - In participants of the CONVERT trial, which enrolled recipients of kidney transplants, conversion of immunosuppressive therapy from calcineurin inhibitors to sirolimus did not improve renal function. More importantly, the intervention was detrimental among patients with impaired kidney function and/or proteinuria. Sirolimus conversion resulted, however, in lower rates of malignancy. PMID- 19556995 TI - Malacoplakia presenting with obstructive nephropathy with bilateral ureter involvement. AB - BACKGROUND: A 48-year-old woman with chronic urinary tract infections presented with uremia. Imaging studies revealed bilateral hydronephrosis and segmental ureteral thickening. INVESTIGATIONS: Physical examination, urine and blood analysis, radiography of the chest, chest CT, abdominal and pelvis CT, renal ultrasonography, cystoscopy, anterograde pyelogram, bladder biopsy. DIAGNOSIS: Malacoplakia involving bilatateral ureters and bladder; cavitary pneumonia. MANAGEMENT: Hemodialysis, placement of bilateral percutaneous nephrostomy tubes, administration of ciprofloxacin, ascorbic acid and bethanechol. PMID- 19556994 TI - Integrating albuminuria and GFR in the assessment of diabetic nephropathy. AB - The evaluation of diabetic nephropathy from research and clinical viewpoints depends on the assessment of two continuous variables, albumin excretion rate (AER) and glomerular filtration rate (GFR). These two parameters form the basis of both the European classification of five stages of diabetic nephropathy, assessed according to changes in AER and GFR (hyperfiltration, normoalbuminuria, microalbuminuria, macroalbuminuria and end-stage renal disease), and the National Kidney Foundation classification of five stages of chronic kidney disease based on categories of estimated GFR. Although increases in AER generally precede a decline in GFR, some patients follow a non-albuminuric pathway to renal impairment. In addition, studies indicate that GFR decreases in a linear fashion from normal or above-normal levels. Whether hyperfiltration is part of the pathogenetic process leading to diabetic nephropathy remains unclear. Ideally, both AER and GFR should be assessed at an early stage in patients being evaluated for diabetic nephropathy. New methods such as the use of cystatin-C-based equations for estimating GFR should be considered because current creatinine based estimates are inaccurate at normal or high GFRs. Serial assessments of both AER and GFR might allow diabetic nephropathy to be diagnosed at early stages of the disease process that are selectively responsive to new interventions. The successful integration of AER categories with the recently defined stages of GFR represents a new challenge in the management of diabetic nephropathy. PMID- 19556996 TI - Weibel-Palade bodies--sentinels of acute stress. AB - Weibel-Palade bodies are uniquely present in endothelial cells and harbor a range of bioactive substances that participate in hemostasis, vasomotion, inflammation and fibrinolysis, in addition to modulating vascular permeability, angiogenic sprouting, and stem cell mobilization. This Perspectives article examines the latest insights into the biogenesis of these organelles and the cellular and molecular mechanisms of their exocytosis. In addition, we advance two hypotheses on the pathogenic role of these organelles: first, in the development of endothelial dysfunction associated with the reduction of nitric oxide bioavailability and accumulation of peroxynitrite and second, as a first-line response to acute stress that determines the balance between regenerative and proinflammatory signals. PMID- 19556999 TI - The SOS response promotes qnrB quinolone-resistance determinant expression. AB - The qnr genes are plasmid-borne fluoroquinolone-resistance determinants widespread in Enterobacteriaceae. Three families of qnr determinants (qnrA, B and S) have been described, but little is known about their expression and regulation. Two new determinants, qnrC and qnrD, have been found recently. Here, we describe the characterization of the qnrB2 promoter and the identification of a LexA-binding site in the promoter region of all qnrB alleles. LexA is the central regulator of the SOS response to DNA damage. We show that qnrB2 expression is regulated through the SOS response in a LexA/RecA-dependent manner, and that it can be induced by the quinolone ciprofloxacin, a known inducer of the SOS system. This is the first description of direct SOS-dependent regulation of an antibiotic-resistance mechanism in response to the antibiotic itself. PMID- 19557000 TI - Ataxia telangiectasia mutated activation by transcription- and topoisomerase I induced DNA double-strand breaks. AB - Ataxia telangiectasia mutated (ATM), the deficiency of which causes a severe neurodegenerative disease, is a crucial mediator for the DNA damage response (DDR). As neurons have high rates of transcription that require topoisomerase I (TOP1), we investigated whether TOP1 cleavage complexes (TOP1cc)-which are potent transcription-blocking lesions-also produce transcription-dependent DNA double strand breaks (DSBs) with ATM activation. We show the induction of DSBs and DDR activation in post-mitotic primary neurons and lymphocytes treated with camptothecin, with the induction of nuclear DDR foci containing activated ATM, gamma-H2AX (phosphorylated histone H2AX), activated CHK2 (checkpoint kinase 2), MDC1 (mediator of DNA damage checkpoint 1) and 53BP1 (p53 binding protein 1). The DSB-ATM-DDR pathway was suppressed by inhibiting transcription and gamma-H2AX signals were reduced by RNase H1 transfection, which removes transcription mediated R-loops. Thus, we propose that Top1cc produce transcription arrests with R-loop formation and generate DSBs that activate ATM in post-mitotic cells. PMID- 19557001 TI - REDD1, an inhibitor of mTOR signalling, is regulated by the CUL4A-DDB1 ubiquitin ligase. AB - The cellular response to hypoxia involves several signalling pathways that mediate adaptation and survival. REDD1 (regulated in development and DNA damage responses 1), a hypoxia-inducible factor-1 target gene, has a crucial role in inhibiting mammalian target of rapamycin complex 1 (mTORC1) signalling during hypoxic stress. However, little is known about the signalling pathways and post translational modifications that regulate REDD1 function. Here, we show that REDD1 is subject to ubiquitin-mediated degradation mediated by the CUL4A-DDB1 ROC1-beta-TRCP E3 ligase complex and through the activity of glycogen synthase kinase 3beta. Furthermore, REDD1 degradation is crucially required for the restoration of mTOR signalling as cells recover from hypoxic stress. Our findings define a mechanism underlying REDD1 degradation and its importance for regulating mTOR signalling. PMID- 19557002 TI - Coiled-coil interactions are required for post-Golgi R-SNARE trafficking. AB - The sorting of post-Golgi R-SNAREs (vesicle-associated membrane protein (VAMP)1, 2, 3, 4, 7 and 8) is still poorly understood. To address this, we developed a system to investigate their localization, trafficking and cell-surface levels. Here, we show that the distribution and internalization of VAMPs 3 and 8 are determined solely through a new conserved mechanism that uses coiled-coil interactions, and that VAMP4 does not require these interactions for its trafficking. We propose that VAMPs 3 and 8 are trafficked while in a complex with Q-SNAREs. We also show that the dileucine motif of VAMP4 is required for both its internalization and retrieval to the trans-Golgi network. However, when the dileucine motif is mutated, the construct can still be internalized potentially through coiled-coil interactions with Q-SNAREs. PMID- 19557003 TI - Morning home blood pressure may be a significant marker of nephropathy in Japanese patients with type 2 diabetes: ADVANCED-J study 1. AB - A 3-year multicenter, prospective, randomized, open-label trial (ADVANCED-J) compared the effect of an increased dose of angiotensin-II receptor blocker (ARB) with that of a maintenance dose of ARB plus calcium channel blocker (amlodipine) on blood pressure (BP) control, nephropathy and atherosclerosis in patients with type 2 diabetes and hypertension in whom the usual ARB dose failed to control BP. A cross-sectional analysis using baseline data was conducted. Of 316 patients (recruited between September 2004 and December 2005), 228 patients were evaluated by multiple regression analysis using two models after randomization and exclusions. Model 1 assessed 13 baseline variables (age, sex, estimated diabetes duration, estimated hypertension duration, HbA1c, brain natriuretic peptide (BNP), high-sensitive C-reactive protein (hsCRP), triglycerides (TGs), total cholesterol (TCHO), diabetic retinopathy (DMR), systolic morning home BP (HBP), diastolic morning HBP and brachial-ankle pulse wave velocity (baPWV)) for correlation with the urinary albumin creatinine excretion rate (UACR). In model 2, systolic and diastolic morning HBP was replaced by systolic and diastolic office BP. The systolic morning HBP and systolic office BP or diastolic morning HBP and diastolic office BP correlations were weak, but significant (r=0.43 and 0.48, respectively). BNP, HbA1c, DMR and estimated diabetes duration were significantly correlated with UACR in both models 1 and 2. Although systolic office BP did not show a significant correlation with UACR in model 2, systolic morning HBP showed a significant correlation with UACR in model 1. Morning HBP, but not office BP, may be a significant marker of nephropathy in Japanese patients with type 2 diabetes. PMID- 19557004 TI - Possible association of tumor necrosis factor receptor 2 gene polymorphism with severe hypertension using the extreme discordant phenotype design. AB - The tumor necrosis factor (TNF)-alpha pathway has a key role in regulating insulin resistance. TNF receptor 2 (TNFR2) is an emerging candidate gene for insulin resistance in essential hypertension. We examined the association of insulin resistance and enhanced TNF pathway with severe hypertension and the association of a microsatellite polymorphism of the TNFR2 gene with severe hypertension. Male severe essential hypertensive patients (HT) with the onset before 60 years of age and with genetic predispositions to hypertension were consecutively enrolled at our outpatient department (N=92). Normotensive men (NT) over 50 years of age were randomly registered from the participants in the annual health check program (N=78). Patients were selected as HT and NT who met stringent criteria for systolic/diastolic blood pressure (SBP/DBP) levels >or=180 and/or 110 mm Hg and <120/80 mm Hg, respectively. HT revealed significantly higher plasma insulin levels, C-reactive protein (CRP) and soluble fraction of TNFR2 concentrations (sTNFR2) than NT. A microsatellite polymorphism of the CA repeat in intron 4 of the TNFR2 gene was analyzed. The allele frequency of CA16 in HT differed significantly from that in NT (66/184 vs. 36/156, P=0.01 by chi(2) analysis). In HT, the CA16 carriers showed significantly higher SBP and plasma insulin levels and a higher tendency of sTNFR2 than did those without this allele. In NT, CA16 carriers revealed significantly higher sTNFR2 and CRP levels than did the CA16 non-carriers. These results suggest that the TNFR2 gene locus has a potential effect on developing severe hypertension through the augmented TNF pathway and insulin resistance. PMID- 19557005 TI - Alcohol sensitivity, alcohol use and hypertension in an older Chinese population: the Guangzhou Biobank Cohort Study. AB - Although the J-shaped association between alcohol consumption and blood pressure (BP) is well known, the effect of alcohol sensitivity on this relationship is less clear. We studied the association of alcohol sensitivity and alcohol use with BP and hypertension. This cross-sectional analysis included 19 335 older participants from the Guangzhou Biobank Cohort Study recruited from 2003 to 2006, using clinically measured BP and self-reported alcohol use and alcohol sensitivity. Alcohol use was rare in women, in whom light-to-moderate drinkers (<140 g ethanol per week) without alcohol sensitivity had lower systolic and diastolic BPs (mean difference 5.3 (95% CI 3.8-6.9) mm Hg and 1.9 (1.1-2.7) mm Hg, respectively) and a reduced risk of hypertension (0.62 (0.53-0.72)) relative to never drinkers. Similarly, excessive drinkers (>or=140 g ethanol per week) without alcohol sensitivity had a significantly higher systolic and diastolic BP and risk of hypertension than did nondrinkers (mean difference 5.1 (2.8-7.4) mm Hg, 2.7 (1.5-4.0) mm Hg and 34% (8-66%), respectively, for men). These differences were even greater for men with alcohol sensitivity (mean differences 12.0 (8.9-15.2) mm Hg, 6.2 (4.5-7.9) mm Hg and 95% CI (46-159%), respectively). Alcohol sensitivity and alcohol use were both associated with elevated BP and risk of hypertension in an older Chinese population. Alcohol sensitivity may aggravate the effect of drinking on BP. Limiting alcohol use to two drinks per day for men and one drink a day for women may be suitable for East Asians. Reduction of alcohol consumption should be an important public health target. PMID- 19557006 TI - Factors associated with uncontrolled hypertension and cardiovascular risk in hypertensive 60-year-old men and women--a population-based study. AB - The objective of this study was to describe cardiovascular risk profiles of 60 year-olds with uncontrolled diagnosed hypertension (>or=140/90 mm Hg) in comparison with individuals with controlled diagnosed hypertension. To study how medical, lifestyle and socioeconomic factors are associated with uncontrolled diagnosed hypertension in men and women separately, a population-based, cross sectional study of 4228 60-year-olds in Sweden, of whom 503 men and 445 women had previously diagnosed hypertension, was conducted. Physical examination including measuring blood pressure was carried out, and a medical/lifestyle/socioeconomic questionnaire was completed. Only 22% of the men and 33% of the women with diagnosed hypertension had a blood pressure below 140/90 mm Hg. Both men and women had multiple cardiovascular risk factors in addition to hypertension. Antihypertensive monotherapy was more common in men and women with controlled hypertension. None of the participants without pharmacological treatment had a controlled blood pressure. Four factors were independently associated with uncontrolled hypertension in men: waist circumference above 97 cm (odds ratio (OR)=1.85, confidence interval (CI)=1.17-2.92), coronary heart disease (CHD) (OR=0.28, CI=0.17-0.46), no health care for financial reasons (OR=2.71, CI=1.09 6.78) and daily intake of fruit (OR=0.59, CI=0.37-0.93). In women, three factors remained independently associated: waist circumference above 78 cm (OR=1.93, CI=1.09-3.43), CHD (OR=0.36, CI=0.18-0.72) and living in an apartment (OR=0.55, CI=0.35-0.85). More efforts are warranted to reduce blood pressure and to modulate associated risk factors to be able to reduce the high morbidity and mortality observed in individuals with hypertension. PMID- 19557007 TI - Nationwide survey of antihypertensive treatment for acute intracerebral hemorrhage in Japan. AB - Acute hypertension is associated with hematoma enlargement and poor clinical outcomes in patients with intracerebral hemorrhage (ICH). However, the method of controlling blood pressure (BP) during the acute phase of ICH remains unknown. The aim of this study is to show current strategies about this issue in Japan. Questionnaires regarding antihypertensive treatment (AHT) strategies were sent to neurosurgeons, neurologists and others responsible for ICH management in 1424 hospitals. Of 600 respondents, 550 (92%) worked at hospitals wherein acute ICH patients are managed and 548 (99.6%) of them agreed with the application of AHT within 24 h of ICH onset. Most answered that the systolic BP threshold for starting AHT was 180 mm Hg (36%) or 160 mm Hg (31%), which differed significantly between neurosurgeons (median, 160 mm Hg) and neurologists/others (180 mm Hg, P<0.001). The goal of lowering systolic BP was to reach a maximum of 140, 150 or 160 mm Hg according to 448 respondents (82%) and 209 (38%) intensively lowered systolic BP to A), one had a 1-bp deletion (c.2029delA), two had 2-bp deletions (c.239_240delGA and c.1670_1671delCA) and one had a 58-bp duplication (c.1138_1195dup). They caused premature termination, resulting in the truncation of the PTCH1 protein. Analysis of a high-density single nucleotide polymorphism (SNP) mapping array showed a large approximately 1.2-Mb deletion, including the PTCH1 gene in one allele, in a family in which PTCH1 mutations were not identified at the sequence level. These data indicated that all the six families who were diagnosed with BCNS had mutations in the PTCH1 gene and that a single copy of a PTCH1 mutation causes BCNS. PMID- 19557016 TI - MTHFR polymorphisms, dietary folate intake and breast cancer risk in Chinese women. AB - To evaluate the relationship between dietary folate intake and genetic polymorphisms of 5,10-methylenetetrahydrofolate reductase (MTHFR) with reference to breast cancer risk, we conducted a case-control study with 669 cases and 682 population-based controls in the Jiangsu Province of China. MTHFR C677T and A1298C genotypes were identified using PCR-RFLP (restrictrion fragment length polymorphism) methods. Dietary folate intake was assessed using an 83-item food frequency questionnaire. Odds ratios (ORs) were estimated with an unconditional logistic model. The frequencies of MTHFR C677T C/C, C/T and T/T genotypes were 32.37, 48.88 and 18.75% in cases and 37.66, 48.24 and 14.10% in controls, respectively. The difference in distribution was significant (chi(2)=6.616, P=0.037), the T/T genotype being associated with an elevated OR (adjusted for age, menopausal status, body mass index (BMI), income, work intensity and status of smoking and drinking) for breast cancer (1.62, 95% confidence interval (95% CI): 1.14-2.30). The frequencies of MTHFR A1298C A/A, A/C and C/C were 71.47, 27.08 and 1.44% in cases and 68.11, 30.13 and 1.76% in controls, respectively, with no significant differences being found (chi(2)=1.716, P=0.424). A significant inverse relationship was observed between folate intake and breast cancer risk. Compared with the lowest tertile of folate intake, the adjusted OR for breast cancer in the top tertile was 0.70 (95% CI: 0.53-0.92). However, no significant interaction was observed between folate intake and the MTHFR C677T polymorphism. Among individuals with the MTHFR A1298C A/A genotype, adjusted ORs for breast cancer were 0.89 (0.62-1.27) and 1.69 (1.20-2.36) for the second to the third tertile of folate intake compared with the highest folate intake group (tread test, P=0.0008). The findings of this study suggest that MTHFR genetic polymorphisms and dietary intake of folate may modify susceptibility to breast cancer. PMID- 19557014 TI - Nedd4 and Nedd4-2: closely related ubiquitin-protein ligases with distinct physiological functions. AB - The Nedd4 (neural precursor cell-expressed developmentally downregulated gene 4) family of ubiquitin ligases (E3s) is characterized by a distinct modular domain architecture, with each member consisting of a C2 domain, 2-4 WW domains, and a HECT-type ligase domain. Of the nine mammalian members of this family, Nedd4 and its close relative, Nedd4-2, represent the ancestral ligases with strong similarity to the yeast, Rsp5. In Saccharomyces cerevisiae Rsp5 has a key role in regulating the trafficking, sorting, and degradation of a large number of proteins in multiple cellular compartments. However, in mammals the Nedd4 family members, including Nedd4 and Nedd4-2, appear to have distinct functions, thereby suggesting that these E3s target specific proteins for ubiquitylation. In this article we focus on the biology and emerging functions of Nedd4 and Nedd4-2, and review recent in vivo studies on these E3s. PMID- 19557017 TI - Allele frequencies of the ABCC11 gene for earwax phenotypes among ancient populations of Hokkaido, Japan. AB - Human earwax is classified into wet and dry types, which are determined by a single-nucleotide polymorphism in the adenosine triphosphate-binding cassette, sub-family C11 (ABCC11) gene locus. To investigate the allele frequencies of the ABCC11 locus within ancient populations on the Northern Japanese island of Hokkaido, amplified product-length polymorphisms were analyzed for 50 specimens of the Okhotsk people and 35 specimens of the Jomon and Epi-Jomon people excavated from various archaeological sites of Hokkaido. Of these specimens, 31 Okhotsk and 19 Jomon/Epi-Jomon samples were genotyped successfully. Frequencies of the wet-type allele in the Jomon/Epi-Jomon people, considered a major ancestor of the Ainu, were higher than those of other Northeastern Asian populations, including the modern Ainu. By contrast, in the Okhotsk people, believed to originate from East Siberia, frequencies of the dry-type allele were relatively higher than those in the Ainu and Jomon/Epi-Jomon people. These results suggest that gene flow from the Northeastern Asian Continent to descendants of the Jomon/Epi-Jomon people of Hokkaido through the Okhotsk people occurred, resulting in the establishment of the Ainu. PMID- 19557018 TI - The effects of hypoxia on the ERG in paediatric cerebral malaria. AB - BACKGROUND/AIMS: Cerebral malaria (CM) is a disease of high mortality worldwide. It can be associated with malarial retinopathy (MR) resulting from impaired perfusion within the retinal microvasculature. Areas of capillary non-perfusion (CNP) appear white (retinal whitening) on ophthalmoloscopy. In this study, electrophysiological investigations were performed to investigate the physiological consequences of these hypoxic and ischaemic changes. METHODS: Children admitted with CM were assessed for inclusion in the study. Those with MR underwent further detailed fundus assessment to quantify retinal whitening and were then designated a severity score. Electrophysiological recordings were performed using a miniganzfeldt stimulator with calibration to the International Society for Clinical Electrophysiology of Visual (ISCEV) standards. ERG data were then analysed with respect to presence of MR and also graded disease severity. RESULTS: Thirty-one children were recruited with a diagnosis of CM, 20 had MR (group 1), and 11 had absent MR (group 2). Statistical analyses of these two groups showed a significant relationship between reduced single flash cone b wave amplitude (CBWA) and increased severity of retinal whitening/CNP (P<0.05). Cone and maximal response b : a wave ratios remained >1 in all subjects. CONCLUSION: Retinal whitening/CNP in MR is associated with significant changes in ERG cone b wave function. The relatively high b : a ratio is compatible with the high frequency of MR resolution without sequelae. PMID- 19557019 TI - High glucose attenuates insulin-induced VEGF expression in bovine retinal microvascular endothelial cells. AB - PURPOSE: To investigate the effect of high glucose on insulin-induced vascular endothelial growth factor (VEGF) expression in bovine retinal microvascular endothelial cells (BRECs) and to probe into related mechanisms. METHODS: BRECs were isolated as primary cultures and identified by immunostaining. Passage cells were initially exposed to normal (5 mM) or high glucose (30 mM) for 3 days, and equimolar L-glucose was supplemented for osmotic equation. BRECs were then treated with 100 nM insulin for 24 h or not, and cells were prepared for the determination of VEGF mRNA expression by real-time PCR. VEGF protein was determined by human umbilical vein endothelial cell proliferation assay, immunofluorescence, and ELISA. BRECs were treated with 5 or 30 mM glucose for 3 days and then cells cultured with 5 mM glucose were exposed to the PI3-K inhibitor wortmannin (100 nM), the P42/44 mitogen-activated protein kinase (MAPK) inhibitor U0126 (50 microM), or to the protein kinase C (PKC) inhibitor GF109203X (2 microM) 1 h before addition of 100 nM insulin. Twenty-four hours after incubation with insulin, the cells were subjected to real-time PCR and ELISA analyses. RESULTS: Insulin or high glucose alone markedly increased VEGF mRNA and protein levels in BRECs (P<0.05, two-way ANOVA). However, the combination of insulin and high glucose displayed a weaker effect in promoting VEGF expression than did insulin alone (P<0.05, t-test). Pretreatment of cells with PI3-K inhibitor significantly (P<0.05, one-way ANOVA) suppressed the insulin-induced VEGF expression; neither pretreatment with the PKC inhibitor nor with the P42/p44 MAPK inhibitor showed an effect on the expression of VEGF at the mRNA or protein level (P>0.05, one-way ANOVA). CONCLUSIONS: Both insulin and high glucose can markedly increase VEGF expression in BRECs at the mRNA and protein level. We propose that insulin may upregulate VEGF expression through the PI3-K signalling pathway in BRECs, and high glucose may attenuate insulin-induced VEGF expression by impairing PI3-K signalling pathways. PMID- 19557020 TI - Paradoxical vascular-fibrotic reaction after intravitreal bevacizumab for retinopathy of prematurity. PMID- 19557021 TI - Poor public health knowledge about glaucoma: fact or fiction? AB - PURPOSE: To document public awareness and knowledge of glaucoma. PATIENTS AND METHODS: The study used health knowledge questionnaires. (a) A short, structured telephone interview was performed with a nationally representative sample of 1009 people. (b) A more detailed questionnaire was administered in two contrasting regions by telephone (500 interviews from the Isle of Wight and 226 interviews from Ealing) or face to face (300 interviews from Ealing). RESULTS: Between 71 and 93% of those interviewed by telephone reported having heard of glaucoma, compared with only 23% of those interviewed face to face in Ealing. Of those who reported having heard of glaucoma, over 80% had at least some knowledge about the disease. CONCLUSION: This is the first study of public awareness of glaucoma across the UK. We found a relatively high level of awareness and knowledge of glaucoma in the general UK population, but identified at least one pocket of poor knowledge in a specific sub-population. PMID- 19557023 TI - Retinal vascular occlusions occur more frequently in the more affected eye in exfoliation syndrome. AB - PURPOSE: To evaluate the relationship between retinal vascular occlusions (RVOs) and exfoliation syndrome (XFS) in a cohort of patients with the two conditions. METHODS: We reviewed the records of patients with XFS with or without glaucoma and any type of RVO between 1983 and 2007. Patients with prior incisional surgery or a history of uveitis were excluded. Data collected included demographics, systemic comorbidities, type of RVO, and intraocular pressure (IOP) before the RVO. Slit-lamp biomicroscopy regarding the presence of exfoliation material on the lens capsule and pupillary margin before the vascular event was used to evaluate the laterality and degree of XFS. RESULTS: We identified 36 patients (mean age 78.4+/-8.3 years, 19 women). Most patients were of European descent (34/36) and 20 (56%) had no prior glaucoma diagnosis. The most common retinal vascular events were central retinal vein occlusion (18/36) and BRVO (10/36). Mean IOP between eyes with (19.5+/-6.5 mm Hg) and without (17.9+/-4.8 mm Hg) RVO was similar (P=0.12). RVOs occurred more commonly in the eye with more pronounced XFS in 92% (33/36) of the cases. A similar agreement was found when considering patients with and without glaucoma separately (94% (15/16) vs90% (18/20); P=0.83). In addition, no difference in the agreement percentage was observed when comparing patients with unilateral XFS (87% (13/15)) with all study patients (P=0.87). CONCLUSIONS: Retinal vascular occlusion in patients with XFS occurs most often in the affected or more severely affected eye. As vascular occlusions happened in patients with and without glaucoma in similar proportions, the presence of XFS seems to play an important role in these findings. PMID- 19557024 TI - Miyake-Apple study of the rotational stability of the Acrysof Toric intraocular lens after experimental eye trauma. AB - PURPOSE: To evaluate the rotational stability of the Acrysof Toric intraocular lens (IOL) after experimental eye trauma. METHODS: Two human cadaver eyes were prepared in accordance with the Miyake-Apple technique, with closed-system modification. After phacoemulsification, an Acrysof Toric IOL was implanted in the capsular bag. Experimental traumatisation of the globe was achieved by pressing against the eye wall using a cotton tip or a fingertip, whereas the IOL was observed from the posterior view. Digital photographs of the eye before and after the procedures were overlaid to detect and measure rotation of the IOL. RESULTS: The IOL rotated 5.80 degrees when the traumatic procedures did not cause important leakage from the incision. When the traumatic procedures caused important leakage from the incision and anterior chamber collapse, the IOL rotated 41.00 degrees. CONCLUSION: Ocular trauma can cause rotation of the Acrysof Toric IOL. In the event of an eye trauma with no or insignificant leakage from the incision, the IOL rotates less than when the trauma causes significant leakage from the incision. PMID- 19557025 TI - Comparison of the ETDRS logMAR, 'compact reduced logMar' and Snellen charts in routine clinical practice. AB - AIM: To compare the performance of the ETDRS logMAR, compact reduced logMAR and Snellen charts in an ophthalmic outpatient setting. METHODS: The reliability and reading times of the charts were compared in a stratified sample of 40 eyes of 40 ophthalmic patients with a variety of stable eye diseases. In order to simulate a clinical setting, forced-choice testing was not used. RESULTS: Similar acuity results were recorded from all three charts, suggesting a lack of a systematic bias as regards chart design. A small practice effect was observed for all charts but was greatest for Snellen and least for ETDRS. The test-retest variability of the charts was similar, with the 95% tolerance limit for change being +/-0.14 logMAR for ETDRS, +/-0.16 for reduced logMAR and +/-0.18 for Snellen. The mean reading times for the subjects were 34.65 s for ETDRS, 21.17 s for reduced logMAR and 18.67 s for Snellen. CONCLUSION: The performance of the compact reduced logMAR chart was intermediate between Snellen and ETDRS. The theoretical advantages of the ETDRS design were still measurable in a clinical setting but the magnitude of the advantage in terms of test-retest reliability was fairly small and the time taken to complete the EDTRS was 1.86 times that of the Snellen chart. PMID- 19557026 TI - Management of inadvertent peribulbar injection of acetazolamide: a case report. PMID- 19557027 TI - Enzymatic synthesis of bis-5-alkylresorcinols by resorcinol-producing type III polyketide synthases. AB - No enzyme systems responsible for the biosynthesis of structurally and biosynthetically intriguing bis-5-alkylresorcinols produced by plants have been identified. Herein, we show that bacterial, fungal and plant alkylresorcinol producing type III polyketide synthases (PKSs), such as ArsB in the Gram-negative bacterium Azotobacter vinelandii, ORAS in the fungus Neurospora crassa and ARAS2 in the rice plant Oryza sativa, can synthesize bis-5-alkylresorcinol from alkanedioic acid N-acetylcysteamine dithioester as a starter substrate and from malonyl-CoA as an extender substrate by two-step conversion. Plants presumably use a type III PKS for the biosynthesis of bis-5-alkylresorcinols. PMID- 19557028 TI - Relationship between peroxisome proliferator-activated receptor-gamma activation and the ameliorative effects of ascochlorin derivatives on type II diabetes. AB - Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a crucial factor in the development of insulin resistance associated with type II diabetes. We previously found that 4-O-carboxymethyl ascochlorin, a derivative of ascochlorin, ameliorates diabetes and activates PPAR-gamma. Here, we compared the relationship between the amelioration of type II diabetes in db/db mice lacking leptin receptor, and PPAR-gamma activation by 4-O-carboxymethyl-ascochlorin, as well as by 4-O-methyl-ascochlorin, a derivative that does not activate PPAR-gamma. Administration of these compounds significantly reduces blood glucose in a dose dependent manner, whereas blood cholesterol is significantly elevated in 4-O carboxymethyl-ascochlorin-treated mice but is significantly decreased in 4-O methyl-ascochlorin-treated mice. Pioglitazone, a potent PPAR-gamma agonist with a thiazolidinedione structure, reduces glucose but elevates cholesterol blood levels. These results suggest that ascochlorin derivatives ameliorate diabetes through a mechanism that is probably independent of PPAR-gamma activation, although PPAR-gamma activation could be partially involved in the ameliorative effect in certain derivatives. PMID- 19557029 TI - Sch 1385568, a new azaphilone from Aspergillus sp. PMID- 19557030 TI - Astaxanthin dirhamnoside, a new astaxanthin derivative produced by a radio tolerant bacterium, Sphingomonas astaxanthinifaciens. PMID- 19557031 TI - An alternative menaquinone biosynthetic pathway operating in microorganisms: an attractive target for drug discovery to pathogenic Helicobacter and Chlamydia strains. AB - Menaquinone is an essential vitamin as an obligatory component of the electron transfer pathway in microorganisms. Menaquinone has been shown to be derived from chorismate by eight enzymes, designated MenA to -H in Escherichia coli. However, bioinformatic analyses of whole-genome sequences have suggested that some microorganisms, such as Helicobacter pylori and Campylobacter jejuni, which are known to cause gastric carcinoma and diarrhea, respectively, do not have orthologs of most of the men genes, although they synthesize menaquinone. The (13)C-labeling pattern of menaquinone purified from Streptomyces coelicolor A3(2) grown on [U-(13)C]glucose was quite different from that of E. coli, suggesting that an alternative pathway was operating in the strain. We searched for candidate genes participating in the alternative pathway by in silico screening, and the involvement of these genes in the pathway was confirmed by gene disruption experiments. We also used mutagenesis to isolate mutants that required menaquinone for their growth and used these mutants as hosts for shotgun cloning experiments. Metabolites that accumulated in the culture broth of mutants were isolated and their structures were determined. Taking these results together, we deduced the outline of the alternative pathway, which branched at chorismate in a similar manner to the known pathway but then followed a completely different pathway. As humans and some useful intestinal bacteria, such as lactobacilli, lack the alternative pathway, it would be an attractive target for the development of chemotherapeutics. PMID- 19557032 TI - NovQ is a prenyltransferase capable of catalyzing the addition of a dimethylallyl group to both phenylpropanoids and flavonoids. AB - NovQ is a member of a recently identified CloQ/NphB class of prenyltransferases. Although NphB has been well characterized as a prenyltransferase with flexibility against aromatic substrates, few studies have been carried out on characterization of NovQ. Hence, in this study, we investigate the kinetics, substrate specificity and regiospecificity of NovQ. The corresponding novQ gene was cloned from Streptomyces niveus, which produces an aminocoumarin antibiotic, novobiocin. Recombinant NovQ was overexpressed in Escherichia coli and purified to homogeneity. The purified enzyme was a soluble monomeric 40-kDa protein that catalyzed the transfer of a dimethylallyl group to 4-hydroxyphenylpyruvate (4 HPP) independently of divalent cations to yield 3-dimethylallyl-4-HPP, an intermediate of novobiocin. Steady-state kinetic constants for NovQ with the two substrates, 4-HPP and dimethylallyl diphosphate, were also calculated. In addition to the prenylation of 4-HPP, NovQ catalyzed carbon-carbon-based and carbon-oxygen-based prenylations of a diverse collection of phenylpropanoids, flavonoids and dihydroxynaphthalenes. Despite its catalytic promiscuity, the NovQ catalyzed prenylation occurred in a regiospecific manner. NovQ is the first reported prenyltransferase capable of catalyzing the transfer of a dimethylallyl group to both phenylpropanoids, such as p-coumaric acid and caffeic acid, and the B-ring of flavonoids. This study shows that NovQ can serve as a useful biocatalyst for the synthesis of prenylated phenylpropanoids and prenylated flavonoids. PMID- 19557033 TI - JBIR-44, a new bromotyrosine compound from a marine sponge Psammaplysilla purpurea. PMID- 19557034 TI - Mutant mouse p53 transgene elevates the chemical induction of tumors that respond to gene silencing with siRNA. AB - To study the role of mutant p53 in the induction and cure of tumors, we generated transgenic mice carrying mutant p53 (mp53) containing a 9 bp deletion in exon 6 in addition to wild-type p53, expressing both p53 and mp53. The mp53 cDNA was cloned from a radiation-induced mouse tumor and ligated to the chicken beta-actin promoter/CMV-IE enhancer in the expression vector. The presence of mp53 suppressed p21 expression in primary fibroblasts after ionizing irradiation, indicating the dominant-negative activity of mp53 in the mice. These mice developed fibrosarcomas after the subcutaneous injection of 3-methylcholanthrene with an incidence 1.7-fold higher than that of wild-type mice (42% excess). The tumors were then treated via a potent atelocollagen delivery system with small interfering RNA (siRNA), that targeted the promoter/enhancer of the expression vector, resulting in the suppression of tumor growth in 30% of 44 autochthonous tumors, including four cures, and their transplants, the total fraction corresponding to the tumor excess. This suppressive effect involved the induction of apoptosis. These results indicate that mp53 activity causes tumors that can be suppressed by subsequent silencing of mp53 in the presence of wild-type p53 alleles. PMID- 19557037 TI - Of hearts and heads. PMID- 19557038 TI - No adverse results. PMID- 19557039 TI - Oblique laterals. PMID- 19557040 TI - Converting clinics. PMID- 19557041 TI - Warranting the effort. PMID- 19557042 TI - Useful contact. PMID- 19557043 TI - Lead poisoning. PMID- 19557044 TI - Shade guides. PMID- 19557045 TI - Swine flu pandemic announced. PMID- 19557047 TI - Focus on world oral health. PMID- 19557050 TI - New health secretary. PMID- 19557048 TI - Teddy bears help anxious children. PMID- 19557053 TI - Tooth decay treatments for children to be examined in national project. PMID- 19557060 TI - Do school break-time policies influence child dental health and snacking behaviours? An evaluation of a primary school programme. AB - OBJECTIVE: The aim of the two-year controlled trial was to evaluate the effectiveness of the 'Boosting Better Breaks' (BBB) break-time policy to reduce obvious decay experience and sugar snacking in a cohort of nine-year-old children attending intervention and control primary schools. STUDY DESIGN: A matched controlled prospective trial design. PARTICIPANTS: Children in Year 5 were invited with their parents/guardians to take part. The children were assessed at baseline and at 24-month follow-up. One hundred and eighty-nine children attended intervention schools and 175 attended control schools which were matched for socio-economic status (SES), school location and co-education status. METHOD: The outcome variables were obvious decay experience and evidence of sugar snacks found in the children's rubbish bags. All children were asked to complete a questionnaire and keep evidence of the snacks they consumed starting from school time break to when they retired for bed in a numbered and coded 'rubbish bag' on a specific collection day at baseline and 24-month follow-up. All children had a dental examination at baseline and 24-month follow-up. RESULTS: Sixty percent of children at baseline and all of the children at follow-up had at least one sugar snack in their rubbish bag. The most popular snacks at follow-up were sweets, chocolate, crisps and carbonated drinks. In the school environment children attending BBB policy schools had significantly lower mean scores for sugar snacks scores at baseline but equivalent mean sugar snacks scores at follow-up compared with children attending control schools. In the outside school environment there was no effect of school intervention on sugar snack scores. Decay into dentine at follow-up was predicted by school intervention status and evidence of sugar snacks consumption outside school and at home. CONCLUSIONS: The BBB break-time policy did not achieve its health promotion goals of promoting child dental health or encouraging children to adopt healthier dietary habits in school or in the wider environment in which they lived. PMID- 19557076 TI - Online Methods. PMID- 19557061 TI - Calcium and vitamin D use among adults in periodontal disease maintenance programmes. AB - OBJECTIVES: To determine the level of calcium and vitamin D oral supplementation in patients in periodontal disease maintenance programmes. DESIGN: Convenience survey. SETTING: St. Louis Metropolitan region. SUBJECTS AND METHODS: Patients (n = 228) in two university-based, periodontal disease maintenance programmes. MAIN OUTCOME MEASURES: Reported amounts of oral calcium and vitamin D supplementation were tested for differences based on gender and race. RESULTS: The last published recommended daily intakes from the United States (US) Food and Nutrition Board (FNB) for adults >50 years of age are 1,200 mg calcium and 400 IU vitamin D (or 600 IU if over 70). The mean age of the 228 patients (125 females and 103 males) was 63.6 +/- 11.0 years (standard deviation). Of the 228 patients surveyed: (1) 204 (89%) were >50 years of age and of these, only 15 (7%) met the US FNB's recommended intakes of calcium and vitamin D from supplementation; (2) 138 (66%) reported that they took no oral supplementation, with significantly more males (n = 82) than females (n = 56) not taking supplementation (p = 0.03); (3) 88 (39%) took calcium supplementation, with females (947 +/- 511 mg/day) taking significantly (p <0.001) more than males (632 +/- 907 mg/day); and (4) 66 (29%) took vitamin D supplementation, with females(420 +/- 227 IU/day) taking approximately the same amount as males (443 +/- 317 IU/day, p >0.05). The amounts of oral supplementation did not vary with race (p >0.05). CONCLUSION: The use of calcium and vitamin D supplementation has been promoted for years, yet the numbers of adults taking supplements remains low and the level of supplementation varies greatly. Knowledge of the benefits of supplementation needs to be better disseminated and research needs to be conducted to determine optimal levels of calcium and vitamin D supplementation. PMID- 19557077 TI - High marks for GWAS. AB - Two genome-wide association studies for testicular cancer report associations at three new loci, including two candidate genes previously implicated in testicular development, KITLG (ligand for the receptor tyrosine kinase) and SPRY4 (sprouty 4). These studies are notable for the high effect sizes detected and the biological plausibility of the candidate genes. PMID- 19557078 TI - TET2 mutations in myelodysplasia and myeloid malignancies. AB - The genetic basis of myelodysplasia has long been enigmatic, with few common targets of mutation known. A new study reports common mutations in the TET2 gene in myelodysplasia and related myeloid malignancies, suggesting that TET2 has an important role in hematopoiesis and in the pathogenesis of this disease. PMID- 19557079 TI - Tumors line up for a letdown. AB - MicroRNAs (miRNAs) and the pathways that regulate their expression have critical functions during normal development. A new study demonstrates that select cancer cells have appropriated one developmental mechanism of miRNA regulation, the inhibition of let-7 biogenesis by the Lin-28 and Lin-28B RNA binding proteins, to rid themselves of an antitumorigenic miRNA. PMID- 19557093 TI - Aripiprazole in the treatment of Huntington's disease: a case series. AB - OBJECTIVES: The aim of the study was to describe the effects of aripiprazole, a new atypical antipsychotic drug that acts as a partial dopamine agonist on motor, behavioral and cognitive functions in patients with genetically confirmed Huntington's disease (HD). METHODS AND RESULTS: Three HD patients were evaluated for Unified Huntington Disease Rating Scale part I and II and Beck Depression Inventory at baseline, after two months and one-year treatment. Aripiprazole effectively controlled involuntary movements and psychiatric symptoms, with effects on cognitive functions. CONCLUSIONS: Our case reports suggest that aripiprazole is well tolerated, remarkably improving some of the motor and behavioral symptoms in patients affected by HD. Randomized, controlled, long-term studies are warranted. PMID- 19557095 TI - Neonatal bilateral lidocaine administration into the ventral hippocampus caused postpubertal behavioral changes: An animal model of neurodevelopmental psychopathological disorders. AB - Our aim was to investigate if neonatal bilateral administration of lidocaine into the ventral hippocampus would cause behavioral changes related to schizophrenia. A neonatal ventral-hippocampal lesion (nVH lesion) was made with lidocaine in Wistar male pups. Two groups were formed, the first received lidocaine (4 mug/0.3 muL) and the second an equal volume of vehicle. At day 35 and 56, both groups were tested for social contact, immobility caused by clamping the neck and dorsal immobility, locomotor activity in an open field, and tail flick (TF) latency after a painful heat stimulus. All animals were then killed. Coronal cuts (7 mum) of the brain were obtained and each brain section was stained with cresyl violet eosin. The animals which received the nVH lesion with lidocaine had decreased social interaction at both ages. The rats with lesions, only at day 58 postnatal, increased their distance traveled and ambulatory time, with a decrease in their nonambulatory and reset time. The rats with lesions had a longer duration of immobility caused by clamping the neck and a longer dorsal immobility at both days 34 and 57 compared to control rats. The lidocaine-treated group spent less time to deflect the tail compared to the control group at postpubertal age. The neonatal bilateral administration of lidocaine into the ventral hippocampus caused some alterations, such as chromatin condensation, nucleolus loss, and cell shrinkage, but glial proliferation was not seen. Neonatal bilateral lidocaine administration into the ventral hippocampus caused postpubertal behavioral changes. PMID- 19557096 TI - Duloxetine for the treatment of generalized anxiety disorder: a review. AB - Approximately 16 million people in the United States suffer from anxiety disorders alone, while another 12 million experience both anxiety and at least one other psychiatric condition. Generalized anxiety disorder (GAD) has lifetime prevalence rates between 5% and 6%. Treatment of GAD is aimed primarily at symptom reduction. Duloxetine, a serotonin norepinephrine reuptake inhibitor (SNRI), received Food and Drug Administration (FDA) approval for the treatment of GAD in 2007. This article reviews the pharmacologic profile and seminal clinical trials associated with the FDA indication of duloxetine for GAD. A literature search performed using PubMed with the keywords "duloxetine", "gad", "generalized anxiety disorder", and "venlafaxine XR" yielded 27 articles. We also focused on papers that pooled data from these seminal studies. Data on file from Eli Lilly were also reviewed, including data from the Eli Lilly website. Based on this search, duloxetine was found to be an FDA-approved treatment option for GAD that has been studied in several double-blind, placebo-controlled clinical trials. This review of duloxetine will help physicians to interpret clinical studies properly and also help them to make an informed decision about which patients are the most appropriate candidates for a trial of duloxetine. PMID- 19557094 TI - Proinflammatory cytokines differentially influence adult hippocampal cell proliferation depending upon the route and chronicity of administration. AB - Disturbances of hippocampal plasticity, including impaired dendritic branching and reductions of neurogenesis, are provoked by stressful insults and may occur in depression. Although corticoids likely contribute to stressor-induced reductions of neurogenesis, other signaling messengers, including pro inflammatory cytokines might also be involved. Accordingly, the present investigation assessed whether three proinflammatory cytokines, namely interleukin-1beta (IL-1beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha) (associated with depression) influenced cellular proliferation within the hippocampus. In this regard, systemic administration of TNF-alpha reduced 5-bromo 2-deoxyuridine (BrdU) labeling within the hippocampus, whereas IL-1beta and IL-6 had no such effect. However, repeated but not a single intra-hippocampal infusion of IL-6 and IL-1beta actually increased cellular proliferation and IL-6 infusion also enhanced microglial staining within the hippocampus. Yet, no changes in doublecortin expression were apparent, suggesting that the cytokine did not influence the birth of cells destined to become neurons. Essentially, the route of administration and chronicity of cytokine administration had a marked influence upon the nature of hippocampal alterations provoked, suggesting that cytokines may differentially regulate hippocampal plasticity in neuropsychiatric conditions. PMID- 19557097 TI - Update on ropinirole in the treatment of Parkinson's disease. AB - Ropinirole is a dopamine agonist, approved for use to treat symptoms of early and advanced Parkinson's disease, is now available in a 24-hour formulation in addition to the immediate release version. This review discusses the mode of action of ropinirole and compares the pharmacokinetics of both formulations. Pivotal studies leading to the approval of both preparations are reviewed in terms of efficacy, dose range and side effects. Patient factors such as compliance are discussed in terms of the place for ropinirole in the armamentarium of Parkinson's disease therapies. PMID- 19557098 TI - Emerging treatments for premature ejaculation: focus on dapoxetine. AB - Premature ejaculation (PE) is a common problem in men worldwide. It has a significant impact on affected men and their partners in terms of self-esteem, dissatisfaction with their sexual relationships, personal distress, and interpersonal difficulty. Psychological therapies may achieve short-term improvements, but there are limited data on the long-term success of these methods. Oral therapy with long-acting selective serotonin reuptake inhibitors (SSRIs) improves intravaginal ejaculatory latency time (IELT), but these agents are designed to be administered daily and may be associated with unwanted sexual side effects and withdrawal symptoms upon abrupt discontinuation. Dapoxetine is a short-acting SSRI that can be taken as needed (prn) by men with PE. It has been studied in five separate multicenter, randomized, double-blind, placebo controlled trials involving more than 6000 men with PE. In four studies that evaluated IELT as an endpoint (N = 4843), dapoxetine 30 and 60 mg prn achieved statistically significant increases in IELT versus placebo. Dapoxetine also showed statistically significant improvements in perceived control over ejaculation, PE-related personal distress, and other patient-reported outcomes in all five trials. Dapoxetine treatment is generally well-tolerated, with low incidences of discontinuation syndrome, sexual dysfunction, and treatment emergent mood symptoms. The most common adverse events with dapoxetine included nausea, diarrhea, headache, dizziness, and somnolence. PMID- 19557100 TI - Ceruloplasmin and superoxide dismutase (SOD1) in heterozygotes for Wilson disease: A case control study. AB - At the time of this study, there were five known patients with Wilson disease (WD) in Iceland. The mutation, a 7-bp deletion in exon 7 on chromosome 13 for WD, is only known in Iceland. In twenty healthy Icelandic heterozygotes for WD and their age- and gender-matched controls, copper concentration in plasma, ceruloplasmin (CP) concentration, CP oxidative activity and CP-specific oxidative activity in serum and superoxide dismutase (SOD1) activity in erythrocytes were determined. The same determinations were done on the five WD patients. There was no significant difference in these parameters between the heterozygotes and the controls, although an inclination toward lower CP determinations and higher SOD1 activity in the heterozygotes was noted. As expected the WD patients were low on the copper and CP parameters, but their SOD1 activity was within the upper normal range. In conclusion, the CP parameters and SOD1 activity are within the normal range in Icelandic heterozygotes for WD, although with a trend toward mild dyshomeostasis. This may indicate subclinical copper retention in the heterozygotes, but a bigger study group is needed to confirm this. PMID- 19557099 TI - Sexual dysfunction in patients with schizophrenia treated with conventional antipsychotics or risperidone. AB - OBJECTIVE: To better understand sexual dysfunction in patients with schizophrenia and its associations with prolactin and reproductive hormones. METHODS: This was a secondary analysis of an open-label, one-day study (N = 402). The primary objective of the study was to assess the prevalence of hyperprolactinemia in patients with schizophrenia who had been treated with conventional antipsychotics or risperidone. Other atypical antipsychotics available at the time of the study were not included due to a more favorable prolactin profile. RESULTS: The majority of patients (59% of females and 60% of males) reported impairment of sexual function. In postmenopausal females, risk of impaired sexual interest was increased by 31% for every 10 ng/ml increase in prolactin (p = 0.035). In males, lower testosterone was associated with higher prolactin (p < 0.001) and with orgasmic (p = 0.004) and ejaculatory dysfunction (p = 0.028). CONCLUSION: These findings suggest that hyperprolactinemia may be associated with sexual dysfunction. They also provide more information on the relationships between prolactin, reproductive hormones, and sexual dysfunction. Sexual dysfunction is an understudied yet important consideration in the treatment of schizophrenia. More attention is warranted in this area as it may provide opportunities for improved quality of life and adherence to treatment for patients. PMID- 19557101 TI - Excited delirium: Consideration of selected medical and psychiatric issues. AB - Excited delirium, sometimes referred to as agitated or excited delirium, is the label assigned to the state of acute behavioral disinhibition manifested in a cluster of behaviors that may include bizarreness, aggressiveness, agitation, ranting, hyperactivity, paranoia, panic, violence, public disturbance, surprising physical strength, profuse sweating due to hyperthermia, respiratory arrest, and death. Excited delirium is reported to result from substance intoxication, psychiatric illness, alcohol withdrawal, head trauma, or a combination of these. This communication reviews the history of the origins of excited delirium, selected research related to its causes, symptoms, management, and the links noted between it and selected medical and psychiatric conditions. Excited delirium involves behavioral and physical symptoms that are also observed in medical and psychiatric conditions such as rhabdomyolysis, neuroleptic malignant syndrome, and catatonia. A useful contribution of this communication is that it links the state of excited delirium to conditions for which there are known and effective medical and psychiatric interventions. PMID- 19557102 TI - Restless legs syndrome: differential diagnosis and management with rotigotine. AB - RLS is a common sleep disorder with distinctive clinical features. The prevalence of RLS in Caucasians and North Americans ranges from 5% to 10%. However, only some of these subjects (almost the 3% of the general population) report being affected by a frequent and severe form of the sleep disorder. RLS is diagnosed clinically by means of four internationally recognized criteria that summarize the main characteristics of the sleep disorder. Besides the essential criteria, supportive and associated features of RLS have been established by experts in order to help physicians treat patients with doubtful symptoms. Several clinical conditions may mimic this sleep disorder. In order to increase the sensibility and specificity of RLS diagnosis, doctors should perform a meticulous patient history and then an accurate physical and neurological examination. Dopamine agonists are recognized as the preferred first-line treatment for RLS. Rotigotine is a non-ergoline dopamine agonist with selectivity for D1, D2 and D3 receptors. The drug is administered via transdermal patches which release rotigotine for 24 hours. Four clinical trials demonstrated that this compound is able to improve RLS symptomatology with few and moderate adverse events. Head to head trials are required to compare the efficacy and tolerability of rotigotine with other dopamine agonists administered via oral intake. Rotigotine has been approved by the FDA and EMEA for Parkinson's disease. For the treatment of moderate to severe idiopathic RLS, rotigotine has been recommended for approval by the EMEA and is under review by the FDA. PMID- 19557104 TI - Combination therapy or monotherapy for the depressed type of schizoaffective disorder. AB - Several studies have demonstrated the effectiveness of adjunctive antidepressant drug therapy to improve the depressive or negative symptoms of schizoaffective disorder, however, monotherapy with atypical antipsychotics may be advantageous. We compared the efficacy and safety of risperidone monotherapy versus combination therapy of haloperidol with sertaline for the acute treatment of schizoaffective disorder, depressed type. This is an open label study of 52 female inpatients randomly assigned to risperidone alone (N = 26) or haloperidol in combination with sertraline (N = 26) for 12 weeks. The mean daily doses of medications were: risperidone: 3.75-3.29 mg/day, haloperidol: 5.35-4.15 mg/day, sertraline: 65.39 133.82 mg/day. Efficacy was measured using clinical rating scales of treatment, safety, and tolerability. Risperidone patients showed statistically significant greater improvement than haloperidol-sertraline patients on efficacy measures including Positive and Negative Syndrome Scale and Clinical Global Impressions rating. A higher number of risperidone patients dropped out of the study early. Fewer adverse events and lesser need for concomitant medications occurred in patients on risperidone. The risperidone group showed better psychological, social and occupational functioning (Global Assessment of Functioning) and higher quality of life (Heinrich's Quality of Life Scale). Risperidone has higher antipsychotic efficacy and tolerability compared with haloperidol-sertraline combination for the acute treatment of schizoaffective disorder, depressed type. Both treatments were comparable in terms of antidepressant efficacy. PMID- 19557103 TI - Postulated vasoactive neuropeptide immunopathology affecting the blood brain/blood-spinal barrier in certain neuropsychiatric fatigue-related conditions: A role for phosphodiesterase inhibitors in treatment? AB - Neuropsychiatric symptoms occur in a number of neurological fatigue-related conditions including multiple sclerosis (MS), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and chronic fatigue syndrome (CFS). These conditions have been attributed variably to neuroinflammatory and neurodegenerative processes. While autoimmune pathology, at least in part, has long been suspected in these conditions proof has been elusive. Autoimmune pathomechanisms affecting the blood-brain barrier (BBB) or blood-spinal barrier (BSB) may predispose the BBB/BSB to 'leakiness' and be a precursor to additional autoimmune events resulting in neuroinflammatory or neurodegenerative processes. The aim of the paper is to postulate immunopathology of the cerebrospinal perivascular compartment involving certain vasoactive neuropeptides, specifically pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP), in the etiology of certain neuropsychiatric fatigue related conditions such as MS, ALS, PD, and CFS. Vasoactive neuropeptides (VNs) such as PACAP and VIP have critical roles as neurotransmitters, vasodilators including perfusion and hypoxia regulators, and immune and nociception modulators. PACAP and VIP are widely distributed in the central nervous system (CNS) and have key roles in CNS blood vessels including maintaining functional integrity of the BBB and BSB. Autoimmunity affecting these VNs would likely have a detrimental effect on BBB and BSB functioning arguably predisposing to further pathological processes. Virchow-Robin spaces (VRS) are perivascular compartments surrounding small vessels within the CNS which contribute to the BBB and BSB integrity and contain PACAP and VIP receptors. Autoimmunity of these receptors would likely affect BBB and VRS function and therefore may contribute to the etiology of these conditions by affecting CNS and immunological homeostasis, including promoting neuropsychological symptomatology. PACAP and VIP, as potent activators of adenylate cyclase (AC), have a key role in cyclic adenosine monophosphate (cAMP) production affecting regulatory T cell (Treg) and other immune functions. Phosphodiesterase enzymes (PDEs) catalyze cAMP and PDE inhibitors (PDEIs) maintain cAMP levels and have proven and well known therapeutic benefit in animal models such as experimental allergic encephalomyelitis (EAE). Therefore PDEIs may have a role in therapy for certain neuropsychiatric fatigue-related conditions. PMID- 19557106 TI - A systematic review of quality of life and weight gain-related issues in patients treated for severe and persistent mental disorders: focus on aripiprazole. AB - Aripiprazole is a relatively novel second-generation antipsychotic belonging to the chemical class of benzisoxazole derivatives and is characterized by a unique pharmacological profile which suggests that the drug acts as a dopamine-serotonin system stabilizer. Whereas all previously available antipsychotics are antagonists at D(2) receptors, aripiprazole is the only available partial agonist at these receptors. Thus, it has been suggested that aripiprazole could be associated with a relatively neutral impact on bodyweight, possibly reducing risks of a detrimental impact on the quality of life that often complicates management for a large number of patients diagnosed with severe and persistent mental disorders (SPMDs) treated chronically with antipsychotic medications. However, data from short- and long-term reviewed studies indicate that the prevalence rate of clinically relevant weight gain during therapy with this drug is similar to that occurring during treatments with other antipsychotic agents, either typical or atypical. Moreover, information on the impact of aripiprazole therapy on the quality of life of patients diagnosed with SPMDs is scarce and characterized by conflicting results. Given these results, further, large, well designed studies are needed before confirming potential advantages of aripiprazole over first-generation antipsychotics and other SGAs. PMID- 19557105 TI - Cognitive impairment after cerebrovascular stroke: Relationship to vascular risk factors. AB - BACKGROUND: Cognitive decline after cerebrovascular stroke has adverse outcome consequences. Since some vascular causes can be prevented and treated, the identification of stroke-related cognitive impairment is a challenge. Patients with cognitive impairment and vascular diseases exhibit higher homocysteine (Hcy) concentrations. Whether Hcy is an independent risk factor for cognitive impairment after stoke is still in question. The objectives of this study were to determine: 1) the relative frequency of first-ever post-stroke dementia (PSD) (three months after onset) in a consecutive sample of our population, 2) the risk factors associated with PSD, and 3) the relationship between Hcy levels and PSD. METHODS: Eighty-one inpatients with first-ever stroke were prospectively evaluated with a neuropsychological battery and event-related evoked potentials (P300) at onset and then after three months. A wide range of demographic, clinical, radiological and laboratory variables were examined. PSD was diagnosed if the clinical presentation fulfilled DSM-IV criteria of vascular dementia, the patient scored /=50% seizure reduction, with respect to baseline, ranged between 37% and 65%. In patients with drug-resistant forms of epilepsy, the percentage of patients reaching a 6-month seizure freedom period was 9%. The percentages of patients who discontinued the experimental drug due to adverse effects ranged between 4% and 24%. Somnolence and dizziness were the most frequently reported adverse effects. Long-term studies demonstrate that ZNS has a good efficacy and tolerability profile, and support its use as adjunctive therapy for epileptic patients. PMID- 19557120 TI - Diabetes mellitus and co-morbid depression: treatment with milnacipran results in significant improvement of both diseases (results from the Austrian MDDM study group). AB - Co-morbid depression is common in patients with diabetes mellitus and has a negative impact on diabetes self-care, adherence to treatment and the development of complications. Effective treatment of depression has been associated with improvement in metabolic parameters. We evaluated the feasibility of a two question screen for co-morbid depression in diabetic patients and studied the effect of the serotonin norepinephrine reuptake inhibitor antidepressant, milnacipran, on metabolic and psychological parameters in 64 type 2 diabetic patients with co-morbid depression. The severity of depression was evaluated using the Beck Depression Inventory (BDI). Patients received milnacipran, and diabetes was treated according to the guidelines of the Austrian Diabetes Association in a 6-month open label study. Metabolic parameters and BDI were measured at baseline and after 1, 3 and 6 months. 46 patients satisfied the criteria for an antidepressant response (reduction of baseline BDI score of at least 50%). Hemoglobin A1c, fasting blood glucose, body mass index, total and LDL cholesterol and serum triglyceride levels were all significantly decreased in these patients at the end of the study whereas in antidepressant non-responders these parameters were not significantly changed. Diagnosis and treatment of depression is an important factor for the improvement of metabolic control in patients with type 2 diabetes and co-morbid depression. PMID- 19557121 TI - Update on the management of symptoms in schizophrenia: focus on amisulpride. AB - Amisulpride is an atypical antipsychotic drug with a unique receptor pharmacology which is dose dependent. It is a standard treatment in dysthymia as well as in psychosis. Amisulpride is efficacious, effective and well tolerated in positive symptoms of schizophrenia: there is extensive evidence that it treats negative symptoms when given in low doses, although relative lack of EPS and an antidepressant effect may contribute. In first-episode patients amisulpride is an option, although there is little comparative work available. Amisulpride has the best evidence as an effective adjunct to clozapine treatment. Regarding intellectual function, amisulpride appears cognitive sparing but the clinical relevance of this remains obscure. There is evidence that amisulpride can improve social function but again there is little comparative work to demonstrate any particular advantages. Regarding the current conventional versus atypical antipsychotic controversy, amisulpride did better in switching studies and meta analyses than in the single large pragmatic randomized trial reported to date. It is a versatile drug, and may offer advantages over other atypical antipsychotic drugs in the treatment of negative and depressive symptoms, and tolerability advantages such as the avoidance of weight gain. Essentially it rests with the treating clinician to employ a rational psychopharmacological approach towards the individual patient: there will be few circumstances in which amisulpride will not be a likely contender as a treatment choice. PMID- 19557122 TI - Treatment and prevention of mania in bipolar I disorder: focus on aripiprazole. AB - Aripiprazole is a second-generation antipsychotic with a unique pharmacologic receptor profile that has efficacy in the treatment and prevention of mania in bipolar I disorder. This article reviews the evidence supporting treatment of adults with bipolar I disorder using aripiprazole as monotherapy or adjunctively during acute mania and its utility as an intramuscular agent for agitation in manic patients. Results from one of the longest bipolar maintenance trials which support aripiprazole as a prophylactic mood stabilizer, specifically against manic relapses, will be discussed as well as a post-hoc analysis that suggests efficacy for rapid cycling bipolar disorder. Safety and tolerability issues, patient-focused perspectives and aripiprazole's place in therapy for bipolar mania will be covered. PMID- 19557124 TI - Establishing a successful bioinformatics core facility team. PMID- 19557123 TI - Current trends in the treatment of infantile spasms. AB - Infantile spasms are an epilepsy syndrome with distinctive features, including age onset during infancy, characteristic epileptic spasms, and specific electroencephalographic patterns (interictal hypsarrhythmia and ictal voltage suppression). Adrenocorticotropic hormone (ACTH) was first employed to treat infantile spasms in 1958, and since then it has been tried in prospective and retrospective studies for infantile spasms. Oral corticosteroids were also used in a few studies for infantile spasms. Variable success in cessation of infantile spasms and normalization of electroencephalograms was demonstrated. However, frequent significant adverse effects are associated with ACTH and oral corticosteroids. Vigabatrin has been used since the 1990s, and shown to be successful in resolution of infantile spasms, especially for infantile spasms associated with tuberous sclerosis. It is associated with visual field constriction, which is often asymptomatic and requires perimetric visual field study to identify. When ACTH, oral corticosteroids, and vigabatrin fail to induce cessation of infantile spasms, other alternative treatments include valproic acid, nitrazepam, pyridoxine, topiramate, zonisamide, lamotrigine, levetiracetam, felbamate, ganaxolone, liposteroid, thyrotropin-releasing hormone, intravenous immunoglobulin and a ketogenic diet. Rarely, infantile spasms in association with biotinidase deficiency, phenylketonuria, and pyridoxine-dependent seizures are successfully treated with biotin, a low phenylalanine diet, and pyridoxine, respectively. For medically intractable infantile spasms, some properly selected patients may have complete cessation of infantile spasms with appropriate surgical treatments. PMID- 19557125 TI - Managing and analyzing next-generation sequence data. PMID- 19557126 TI - Quantification of normal cell fraction and copy number neutral LOH in clinical lung cancer samples using SNP array data. AB - BACKGROUND: Technologies based on DNA microarrays have the potential to provide detailed information on genomic aberrations in tumor cells. In practice a major obstacle for quantitative detection of aberrations is the heterogeneity of clinical tumor tissue. Since tumor tissue invariably contains genetically normal stromal cells, this may lead to a failure to detect aberrations in the tumor cells. PRINCIPAL FINDING: Using SNP array data from 44 non-small cell lung cancer samples we have developed a bioinformatic algorithm that accurately models the fractions of normal and tumor cells in clinical tumor samples. The proportion of normal cells in combination with SNP array data can be used to detect and quantify copy number neutral loss-of-heterozygosity (CNNLOH) in the tumor cells both in crude tumor tissue and in samples enriched for tumor cells by laser capture microdissection. CONCLUSION: Genome-wide quantitative analysis of CNNLOH using the CNNLOH Quantifier method can help to identify recurrent aberrations contributing to tumor development in clinical tumor samples. In addition, SNP array based analysis of CNNLOH may become important for detection of aberrations that can be used for diagnostic and prognostic purposes. PMID- 19557127 TI - Decoupling internalization, acidification and phagosomal-endosomal/lysosomal fusion during phagocytosis of InlA coated beads in epithelial cells. AB - BACKGROUND: Phagocytosis has been extensively examined in 'professional' phagocytic cells using pH sensitive dyes. However, in many of the previous studies, a separation between the end of internalization, beginning of acidification and completion of phagosomal-endosomal/lysosomal fusion was not clearly established. In addition, very little work has been done to systematically examine phagosomal maturation in 'non-professional' phagocytic cells. Therefore, in this study, we developed a simple method to measure and decouple particle internalization, phagosomal acidification and phagosomal endosomal/lysosomal fusion in Madin-Darby Canine Kidney (MDCK) and Caco-2 epithelial cells. METHODOLOGY/PRINCIPAL FINDINGS: Our method was developed using a pathogen mimetic system consisting of polystyrene beads coated with Internalin A (InlA), a membrane surface protein from Listeria monocytogenes known to trigger receptor-mediated phagocytosis. We were able to independently measure the rates of internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in epithelial cells by combining the InlA-coated beads (InlA-beads) with antibody quenching, a pH sensitive dye and an endosomal/lysosomal dye. By performing these independent measurements under identical experimental conditions, we were able to decouple the three processes and establish time scales for each. In a separate set of experiments, we exploited the phagosomal acidification process to demonstrate an additional, real-time method for tracking bead binding, internalization and phagosomal acidification. CONCLUSIONS/SIGNIFICANCE: Using this method, we found that the time scales for internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion ranged from 23-32 min, 3-4 min and 74-120 min, respectively, for MDCK and Caco-2 epithelial cells. Both the static and real-time methods developed here are expected to be readily and broadly applicable, as they simply require fluorophore conjugation to a particle of interest, such as a pathogen or mimetic, in combination with common cell labeling dyes. As such, these methods hold promise for future measurements of receptor-mediated internalization in other cell systems, e.g. pathogen-host systems. PMID- 19557128 TI - Probing natural killer cell education by Ly49 receptor expression analysis and computational modelling in single MHC class I mice. AB - Murine natural killer (NK) cells express inhibitory Ly49 receptors for MHC class I molecules, which allows for "missing self" recognition of cells that downregulate MHC class I expression. During murine NK cell development, host MHC class I molecules impose an "educating impact" on the NK cell pool. As a result, mice with different MHC class I expression display different frequency distributions of Ly49 receptor combinations on NK cells. Two models have been put forward to explain this impact. The two-step selection model proposes a stochastic Ly49 receptor expression followed by selection for NK cells expressing appropriate receptor combinations. The sequential model, on the other hand, proposes that each NK cell sequentially expresses Ly49 receptors until an interaction of sufficient magnitude with self-class I MHC is reached for the NK cell to mature. With the aim to clarify which one of these models is most likely to reflect the actual biological process, we simulated the two educational schemes by mathematical modelling, and fitted the results to Ly49 expression patterns, which were analyzed in mice expressing single MHC class I molecules. Our results favour the two-step selection model over the sequential model. Furthermore, the MHC class I environment favoured maturation of NK cells expressing one or a few self receptors, suggesting a possible step of positive selection in NK cell education. Based on the predicted Ly49 binding preferences revealed by the model, we also propose, that Ly49 receptors are more promiscuous than previously thought in their interactions with MHC class I molecules, which was supported by functional studies of NK cell subsets expressing individual Ly49 receptors. PMID- 19557129 TI - Linoleic acid-induced mitochondrial Ca(2+) efflux causes peroxynitrite generation and protein nitrotyrosylation. AB - It is well known that excessive non-esterified fatty acids in diabetes contribute to the pathogenesis of renal complications although the mechanism remains elusive. Enhanced oxidative stress has been hypothesized as a unified factor contributing to diabetic complications and increased protein nitrotyrosylation has been reported in the kidneys of diabetic patients. In the current manuscript we described that linoleic acid (LA) caused mitochondrial Ca(2+) efflux and peroxynitrite production, along with increased nitrotyrosine levels of cellular proteins in primary human mesangial cells. The peroxynitrite production by LA was found to depend on mitochondrial Ca(2+) efflux. Downregulation of hsp90beta1, which has been previously shown to be essential for polyunsaturated fatty acid induced mitochondrial Ca(2+) efflux, significantly diminished LA-responsive mitochondrial Ca(2+) efflux and the coupled peroxynitrite generation, implicating a critical role of hsp90beta1 in the LA responses. Our results further demonstrated that mitochondrial complexes I and III were directly involved in the LA-induced peroxynitrite generation. Using the well established type 2 diabetic animal model db/db mice, we observed a dramatically enhanced LA responsive mitochondrial Ca(2+) efflux and protein nitrotyrosylation in the kidney. Our study thus demonstrates a cause-effect relationship between LA and peroxynitrite or protein nitrotyrosylation and provides a novel mechanism for lipid-induced nephropathy in diabetes. PMID- 19557130 TI - Severe human influenza infections in Thailand: oseltamivir treatment and risk factors for fatal outcome. AB - BACKGROUND: Influenza is often not recognized as an important cause of severe or fatal disease in tropical and subtropical countries in Southeast Asia. The extent to which Oseltamivir treatment may protect against a fatal outcome in severe influenza infections is not known. Thailand's National Avian Influenza Surveillance (NAIS) system affords a unique opportunity to describe the epidemiology of laboratory-confirmed severe and fatal human influenza infections. METHODOLOGY/PRINCIPAL FINDINGS: During January 2004 through December 2006, 11,641 notifications to the NAIS were investigated in 73 of 76 Thai provinces. Clinical and demographic data and respiratory swab specimens were collected and tested by PCR for influenza. Using the NAIS database, we identified all patients with laboratory confirmed human influenza (A/H3N2, A/H1N1 and Type B) infection. A retrospective medical record review was conducted on all fatal cases with laboratory confirmed influenza and from a sample of hospitalized cases in 28 provinces. The association of underlying risk factors, Oseltamivir treatment and risk of a fatal outcome were examined. Human influenza infections were identified in 2,075 (18%) cases. Twenty-two (1%) deaths occurred including seven deaths in children less than ten years of age. Thirty-five percent of hospitalized human influenza infections had chest X-ray confirmed pneumonia. Current or former smoking; advanced age, hypertension and underlying cardiovascular, pulmonary or endocrine disease were associated with a fatal outcome from human influenza infection. Treatment with Oseltamivir was statistically associated with survival with a crude OR of .11 (95% CI: 0.04-0.30) and .13 (95% CI: 0.04-0.40) after controlling for age. CONCLUSIONS: Severe and fatal human influenza infections were commonly identified in the NAIS designed to identify avian A/H5N1 cases. Treatment with Oseltamivir is associated with survival in hospitalized human influenza pneumonia patients. PMID- 19557131 TI - Detection of the antiviral drug oseltamivir in aquatic environments. AB - Oseltamivir (Tamiflu) is the most important antiviral drug available and a cornerstone in the defence against a future influenza pandemic. Recent publications have shown that the active metabolite, oseltamivir carboxylate (OC), is not degraded in sewage treatment plants and is also persistent in aquatic environments. This implies that OC will be present in aquatic environments in areas where oseltamivir is prescribed to patients for therapeutic use. The country where oseltamivir is used most is Japan, where it is used to treat seasonal flu. We measured the levels of OC in water samples from the Yodo River system in the Kyoto and Osaka prefectures, Japan, taken before and during the flu season 2007/8. No OC was detected before the flu-season but 2-58 ng L(-1) was detected in the samples taken during the flu season. This study shows, for the first time, that low levels of oseltamivir can be found in the aquatic environment. Therefore the natural reservoir of influenza virus, dabbling ducks, is exposed to oseltamivir, which could promote the evolution of viral resistance. PMID- 19557132 TI - High fat feeding induces hepatic fatty acid elongation in mice. AB - BACKGROUND: High-fat diets promote hepatic lipid accumulation. Paradoxically, these diets also induce lipogenic gene expression in rodent liver. Whether high expression of these genes actually results in an increased flux through the de novo lipogenic pathway in vivo has not been demonstrated. METHODOLOGY/PRINCIPAL FINDINGS: To interrogate this apparent paradox, we have quantified de novo lipogenesis in C57Bl/6J mice fed either chow, a high-fat or a n-3 polyunsaturated fatty acid (PUFA)-enriched high-fat diet. A novel approach based on mass isotopomer distribution analysis (MIDA) following 1-(13)C acetate infusion was applied to simultaneously determine de novo lipogenesis, fatty acid elongation as well as cholesterol synthesis. Furthermore, we measured very low density lipoprotein-triglyceride (VLDL-TG) production rates. High-fat feeding promoted hepatic lipid accumulation and induced the expression of lipogenic and cholesterogenic genes compared to chow-fed mice: induction of gene expression was found to translate into increased oleate synthesis. Interestingly, this higher lipogenic flux (+74 microg/g/h for oleic acid) in mice fed the high-fat diet was mainly due to an increased hepatic elongation of unlabeled palmitate (+66 microg/g/h) rather than to elongation of de novo synthesized palmitate. In addition, fractional cholesterol synthesis was increased, i.e. 5.8+/-0.4% vs. 8.1+/-0.6% for control and high fat-fed animals, respectively. Hepatic VLDL-TG production was not affected by high-fat feeding. Partial replacement of saturated fat by fish oil completely reversed the lipogenic effects of high-fat feeding: hepatic lipogenic and cholesterogenic gene expression levels as well as fatty acid and cholesterol synthesis rates were normalized. CONCLUSIONS/SIGNIFICANCE: High-fat feeding induces hepatic fatty acid synthesis in mice, by chain elongation and subsequent desaturation rather than de novo synthesis, while VLDL TG output remains unaffected. Suppression of lipogenic fluxes by fish oil prevents from high fat diet-induced hepatic steatosis in mice. PMID- 19557133 TI - Estimating the incidence of symptomatic rotavirus infections: a systematic review and meta-analysis. AB - BACKGROUND: We conducted for the first time a systematic review, including a meta analysis, of the incidence of symptomatic rotavirus (RV) infections, because (1) it was shown to be an influential factor in estimating the cost-effectiveness of RV vaccination, (2) multiple community-based studies assessed it prospectively, (3) previous studies indicated, inconclusively, it might be similar around the world. METHODOLOGY: Pubmed (which includes Medline) was searched for surveys assessing prospectively symptomatic (diarrheal) episodes in a general population and situation, which also reported on the number of the episodes being tested RV+ and on the persons and the time period observed. A bias assessment tool was developed and used according to Cochrane guidelines by 4 researchers with different backgrounds. Heterogeneity was explored graphically and by comparing fits of study-homogenous 'fixed effects' and -heterogeneous 'random effects' models. Data were synthesized using these models. Sensitivity analysis for uncertainty regarding data abstraction, bias assessment and included studies was performed. PRINCIPAL FINDINGS: Variability between the incidences obtained from 20 studies is unlikely to be due to study groups living in different environments (tropical versus temperate climate, slums versus middle-class suburban populations), nor due to the year the study was conducted (from 1967 to 2003). A random effects model was used to incorporate unexplained heterogeneity and resulted in a global incidence estimate of 0.31 [0.19; 0.50] symptomatic RV infections per personyear of observation for children below 2 years of age, and of 0.24 [0.17; 0.34] when excluding the extreme high value of 0.84 reported for Mayan Indians in Guatemala. Apart from the inclusion/exclusion of the latter study, results were robust. CONCLUSIONS/SIGNIFICANCE: Rather than assumptions based on an ad-hoc selection of one or two studies, these pooled estimates (together with the measure for variability between populations) should be used as an input in future cost-effectiveness analyses of RV vaccination. PMID- 19557134 TI - When stress predicts a shrinking gene pool, trading early reproduction for longevity can increase fitness, even with lower fecundity. AB - BACKGROUND: Stresses like dietary restriction or various toxins increase lifespan in taxa as diverse as yeast, Caenorhabditis elegans, Drosophila and rats, by triggering physiological responses that also tend to delay reproduction. Food odors can reverse the effects of dietary restriction, showing that key mechanisms respond to information, not just resources. Such environmental cues can predict population trends, not just individual prospects for survival and reproduction. When population size is increasing, each offspring produced earlier makes a larger proportional contribution to the gene pool, but the reverse is true when population size is declining. PRINCIPAL FINDINGS: We show mathematically that natural selection can favor facultative delay in reproduction when environmental cues predict a decrease in total population size, even if lifetime fecundity decreases with delay. We also show that increased reproduction from waiting for better conditions does not increase fitness (proportional representation) when the whole population benefits similarly. CONCLUSIONS: We conclude that the beneficial effects of stress on longevity (hormesis) in diverse taxa are a side effect of delaying reproduction in response to environmental cues that population size is likely to decrease. The reversal by food odors of the effects of dietary restriction can be explained as a response to information that population size is less likely to decrease, reducing the chance that delaying reproduction will increase fitness. PMID- 19557135 TI - A systems immunology approach to the host-tumor interaction: large-scale patterns of natural autoantibodies distinguish healthy and tumor-bearing mice. AB - Traditionally, immunology has considered a meaningful antibody response to be marked by large amounts of high-affinity antibodies reactive with the specific inciting antigen; the detection of small amounts of low-affinity antibodies binding to seemingly unrelated antigens has been considered to be beneath the threshold of immunological meaning. A systems-biology approach to immunology, however, suggests that large-scale patterns in the antibody repertoire might also reflect the functional state of the immune system. To investigate such global patterns of antibodies, we have used an antigen-microarray device combined with informatic analysis. Here we asked whether antibody-repertoire patterns might reflect the state of an implanted tumor. We studied the serum antibodies of inbred C57BL/6 mice before and after implantation of syngeneic 3LL tumor cells of either metastatic or non-metastatic clones. We analyzed patterns of IgG and IgM autoantibodies binding to over 300 self-antigens arrayed on slides using support vector machines and genetic algorithm techniques. We now report that antibody patterns, but not single antibodies, were informative: 1) mice, even before tumor implantation, manifest both individual and common patterns of low-titer natural autoantibodies; 2) the patterns of these autoantibodies respond to the growth of the tumor cells, and can distinguish between metastatic and non-metastatic tumor clones; and 3) curative tumor resection induces dynamic changes in these low titer autoantibody patterns. The informative patterns included autoantibodies binding to self-molecules not known to be tumor-associated antigens (including insulin, DNA, myosin, fibrinogen) as well as to known tumor-associated antigens (including p53, cytokeratin, carbonic anhydrases, tyrosinase). Thus, low-titer autoantibodies that are not the direct products of tumor-specific immunization can still generate an immune biomarker of the body-tumor interaction. System-wide profiling of autoantibody repertoires can be informative. PMID- 19557136 TI - Molecular characterization and tissue localization of an F-box only protein from silkworm, Bombyx mori. AB - The eukaryotic F-box protein family is characterized by an F-box motif that has been shown to be critical for the controlled degradation of regulatory proteins. We identified a gene encoding an F-box protein from a cDNA library of silkworm pupae, which has an ORF of 1821 bp, encoding a predicted 606 amino acids. Bioinformatic analysis on the amino acid sequence shows that BmFBXO21 has a low degree of similarity to proteins from other species, and may be related to the regulation of cell-cycle progression. We have detected the expression pattern of BmFBXO21 mRNA and protein and performed immunohistochemistry at three different levels. Expression was highest in the spinning stage, and in the tissues of head, epidermis, and genital organs. PMID- 19557138 TI - Synthesis, characterization, and biological activity studies of copper(II) mixed compound with histamine and nalidixic acid. AB - A mixed copper complex with deprotonated nalidixic acid (nal) and histamine (hsm) was synthesized and characterized by FTIR, UV-Vis, elemental analysis, and conductivity. The crystal structure of [Cu(hsm)(nal)H(2)O]Cl.3H(2)O (chn) showed a pentacoordinated cooper(II) in a square pyramidal geometry surrounded by two N atoms from hsm, two O atoms from the quinolone, and one apical water oxygen. Alteration of bacterial DNA structure and/or associated functions in vivo by [Cu(hsm)(nal)H(2)O]Cl.3H(2)O was demonstrated by the induction of a recA-lacZ fusion integrated at the amyE locus of a recombinant Bacillus subtilis strain. Results from circular dichroism and denaturation of calf thymus DNA (CT-DNA) suggested that increased amounts of copper complex were able to stabilize the double helix of DNA in vitro mainly by formation of hydrogen bonds between chn and the sugars of DNA minor groove. In vivo and in vitro biological activities of the chn complex were compared with the chemical nuclease [Cu(phen)(nal)H(2)O]NO(3) . 3H(2)O (cpn) where phen is phenanthroline. PMID- 19557137 TI - The neuroelectromagnetic inverse problem and the zero dipole localization error. AB - A tomography of neural sources could be constructed from EEG/MEG recordings once the neuroelectromagnetic inverse problem (NIP) is solved. Unfortunately the NIP lacks a unique solution and therefore additional constraints are needed to achieve uniqueness. Researchers are then confronted with the dilemma of choosing one solution on the basis of the advantages publicized by their authors. This study aims to help researchers to better guide their choices by clarifying what is hidden behind inverse solutions oversold by their apparently optimal properties to localize single sources. Here, we introduce an inverse solution (ANA) attaining perfect localization of single sources to illustrate how spurious sources emerge and destroy the reconstruction of simultaneously active sources. Although ANA is probably the simplest and robust alternative for data generated by a single dominant source plus noise, the main contribution of this manuscript is to show that zero localization error of single sources is a trivial and largely uninformative property unable to predict the performance of an inverse solution in presence of simultaneously active sources. We recommend as the most logical strategy for solving the NIP the incorporation of sound additional a priori information about neural generators that supplements the information contained in the data. PMID- 19557139 TI - Optimized null model for protein structure networks. AB - Much attention has recently been given to the statistical significance of topological features observed in biological networks. Here, we consider residue interaction graphs (RIGs) as network representations of protein structures with residues as nodes and inter-residue interactions as edges. Degree-preserving randomized models have been widely used for this purpose in biomolecular networks. However, such a single summary statistic of a network may not be detailed enough to capture the complex topological characteristics of protein structures and their network counterparts. Here, we investigate a variety of topological properties of RIGs to find a well fitting network null model for them. The RIGs are derived from a structurally diverse protein data set at various distance cut-offs and for different groups of interacting atoms. We compare the network structure of RIGs to several random graph models. We show that 3-dimensional geometric random graphs, that model spatial relationships between objects, provide the best fit to RIGs. We investigate the relationship between the strength of the fit and various protein structural features. We show that the fit depends on protein size, structural class, and thermostability, but not on quaternary structure. We apply our model to the identification of significantly over-represented structural building blocks, i.e., network motifs, in protein structure networks. As expected, choosing geometric graphs as a null model results in the most specific identification of motifs. Our geometric random graph model may facilitate further graph-based studies of protein conformation space and have important implications for protein structure comparison and prediction. The choice of a well-fitting null model is crucial for finding structural motifs that play an important role in protein folding, stability and function. To our knowledge, this is the first study that addresses the challenge of finding an optimized null model for RIGs, by comparing various RIG definitions against a series of network models. PMID- 19557140 TI - Management of obsessive-compulsive disorder with fluvoxamine extended release. AB - The pharmacodynamic properties of fluvoxamine maleate include the modulation of different populations of serotonergic, dopaminergic, and sigma receptors and/or transporters, a complex pattern of activity that may account for its efficacy in the treatment of obsessive-compulsive disorder (OCD). Nevertheless, its pharmacokinetic profile and its pattern of side effects may hinder a rapid dose escalation, a therapeutic strategy that might be utterly desirable in patients with OCD. In preclinical studies, the maximum plasma concentration and bioavailability of an extended-release (CR) formulation of fluvoxamine were, respectively, 38% and 16% lower than those of the standard (ie, non-CR) formulation. Recently, the US Food and Drug Administration approved the fluvoxamine CR formulation for the treatment of OCD in adults. This approval was based on the results of a double-blind, placebo-controlled study with 253 OCD patients in which fluvoxamine CR showed a consistently earlier onset of therapeutic effects than other selective serotonin reuptake inhibitors, as reported in previous studies. The use of the CR formulation of fluvoxamine allowed a particularly aggressive dosing strategy at the beginning of the titration phase, ie, treatment could be started with a single dose of fluvoxamine CR 100 mg at bedtime, while keeping the occurrence of side effects and the rate of compliance at levels comparable to those reported for the use of immediate release fluvoxamine. PMID- 19557141 TI - Frontal brain dysfunction in alcoholism with and without antisocial personality disorder. AB - Alcoholism and antisocial personality disorder (ASPD) often are comorbid conditions. Alcoholics, as well as nonalcoholic individuals with ASPD, exhibit behaviors associated with prefrontal brain dysfunction such as increased impulsivity and emotional dysregulation. These behaviors can influence drinking motives and patterns of consumption. Because few studies have investigated the combined association between ASPD and alcoholism on neuropsychological functioning, this study examined the influence of ASPD symptoms and alcoholism on tests sensitive to frontal brain deficits. The participants were 345 men and women. Of them, 144 were abstinent alcoholics (66 with ASPD symptoms), and 201 were nonalcoholic control participants (24 with ASPD symptoms). Performances among the groups were examined with Trails A and B tests, the Wisconsin Card Sorting Test, the Controlled Oral Word Association Test, the Ruff Figural Fluency Test, and Performance subtests of the Wechsler Adult Intelligence Scale. Measures of affect also were obtained. Multiple regression analyses showed that alcoholism, specific drinking variables (amount and duration of heavy drinking), and ASPD were significant predictors of frontal system and affective abnormalities. These effects were different for men and women. The findings suggested that the combination of alcoholism and ASPD leads to greater deficits than the sum of each. PMID- 19557142 TI - A randomized controlled trial of quetiapine for psychosis in Parkinson's disease. AB - INTRODUCTION: Psychosis (delusions and/or hallucinations) is a well-recognized complication of treatment of Parkinson's disease (PD). Quetiapine is a currently favored treatment, but data on its efficacy are equivocal. This trial aimed to provide further evidence on the efficacy of quetiapine in PD psychosis. METHODS: We conducted a 12 week double blind randomized placebo-controlled trial. Time to dropout due to lack of improvement of psychosis was the primary outcome measure. Other important secondary outcomes were evaluated using standard rating scales for PD and psychiatric symptoms. RESULTS: Twenty-four eligible subjects gave consent. The primary outcome, time to dropout, was examined using survival analysis. It was shown that patients in the quetiapine group dropped out earlier than those in the placebo group, but this difference was not significant (p = 0.68). No significant changes were found for any of the secondary outcome measures in either group. CONCLUSIONS: In this study, quetiapine at doses of up to 150 mg/day failed to significantly improve psychosis compared to placebo, however the small sample size does not allow any conclusive interpretation of the results. Quetiapine did not appear to worsen PD motor functioning, but its use was limited by a faster drop out compared with placebo. Significant impediments were difficulty with recruitment and natural fluctuation in symptoms during the trial. PMID- 19557143 TI - Review of teriflunomide and its potential in the treatment of multiple sclerosis. AB - In the light of new cases of progressive multifocal leukoencephalopathy and induced autoimmunity in multiple sclerosis (MS) patients who received treatment with upcoming disease-modifying immunosuppressant drugs with a highly specific mode of action such as natalizumab, rituximab, or alemtuzumab, alternative oral treatment options for a subgroup of less severely affected MS patients are a major focus of drug development. These agents are currently investigated in phase III clinical trials and some of them are characterized by a favorable safety profile. With an emphasis on teriflunomide, the active metabolite of an immunosuppressant approved for the treatment of rheumatoid arthritis since 1998, a number of oral treatment options for patients with MS are discussed. PMID- 19557144 TI - New approaches in the management of insomnia: weighing the advantages of prolonged-release melatonin and synthetic melatoninergic agonists. AB - Hypnotic effects of melatonin and melatoninergic drugs are mediated via MT(1) and MT(2) receptors, especially those in the circadian pacemaker, the suprachiasmatic nucleus, which acts on the hypothalamic sleep switch. Therefore, they differ fundamentally from GABAergic hypnotics. Melatoninergic agonists primarily favor sleep initiation and reset the circadian clock to phases allowing persistent sleep, as required in circadian rhythm sleep disorders. A major obstacle for the use of melatonin to support sleep maintenance in primary insomnia results from its short half-life in the circulation. Solutions to this problem have been sought by developing prolonged-release formulations of the natural hormone, or melatoninergic drugs of longer half-life, such as ramelteon, tasimelteon and agomelatine. With all these drugs, improvements of sleep are statistically demonstrable, but remain limited, especially in primary chronic insomnia, so that GABAergic drugs may be indicated. Melatoninergic agonists do not cause next-day hangover and withdrawal effects, or dependence. They do not induce behavioral changes, as sometimes observed with z-drugs. Despite otherwise good tolerability, the use of melatoninergic drugs in children, adolescents, and during pregnancy has been a matter of concern, and should be avoided in autoimmune diseases and Parkinsonism. Problems and limits of melatoninergic hypnotics are compared. PMID- 19557145 TI - Physico-chemical characteristics of lipoplexes influence cell uptake mechanisms and transfection efficacy. AB - BACKGROUND: Formulation of DNA/cationic lipid complexes (lipoplexes) designed for nucleic acid delivery mostly results in positively charged particles which are thought to enter cells by endocytosis. We recently developed a lipoplex formulation called Neutraplex that allows preparation of both cationic and anionic stable complexes with similar lipid content and ultrastructure. METHODOLOGY/PRINCIPAL FINDINGS: To assess whether the global net charge could influence cell uptake and activity of the transported oligonucleotides (on), we prepared lipoplexes with positive and negative charges and compared: (i) their physicochemical properties by zeta potential analysis and dynamic light scattering, (ii) their cell uptake by fluorescence microscopy and flow cytometry, and (iii) the biological activity of the transported ON using a splicing correction assay. We show that positively or negatively charged lipoplexes enter cells cells using both temperature-dependent and -independent uptake mechanisms. Specifically, positively charged lipoplexes predominantly use a temperature dependent transport when cells are incubated OptiMEM medium. Anionic lipoplexes favour an energy-independent transport and show higher ON activity than cationic lipoplexes in presence of serum. However, lipoplexes with high positive global net charge and OptiMEM medium give the highest uptake and ON activity levels. CONCLUSIONS: These findings suggest that, in addition to endocytosis, lipoplexes may enter cell via a temperature-independent mechanism, which could be mediated by lipid mixing. Such characteristics might arise from the specific lipoplex ultrastructure and should be taken into consideration when developing lipoplexes designed for in vivo or ex vivo nucleic acid transfer. PMID- 19557147 TI - The link between human and transgenic animal studies involving postprandial hypertriglyceridemia and CETP gene polymorphisms. PMID- 19557148 TI - Quantitative evaluation of the amount of delayed myocardial enhancement as a predictor of systolic dysfunction. AB - 30 patients with delayed contrast enhancement in patterns suggestive of myocardial infarctions were reviewed. Infarct mass was quantitatively measured using short axis images obtained in the delayed phase of gadopentetate administration. Left ventricular mass and ejection fraction were measured using short axis, steady state free precession images. A relationship is drawn between increased mass of infarction and decreased left ventricular ejection fraction. For each gram of infarct, there is a 0.5 % reduction in ejection fraction (EF = 50 - (0.48 x gm infarcted myocardium); r2= 0.49). For each % increase of infarcted myocardium, there is a 0.67 % reduction in ejection fraction (EF = 50 - (0.67 x percent of infarcted myocardium); r2= 0.39). Left ventricular ejection fraction correlates inversely with the mass of myocardium with delayed enhancement on cardiac MRI. PMID- 19557146 TI - Skin-derived TSLP triggers progression from epidermal-barrier defects to asthma. AB - Asthma is a common allergic lung disease frequently affecting individuals with a prior history of eczema/atopic dermatitis (AD); however, the mechanism underlying the progression from AD to asthma (the so-called "atopic march") is unclear. Here we show that, like humans with AD, mice with skin-barrier defects develop AD-like skin inflammation and are susceptible to allergic asthma. Furthermore, we show that thymic stromal lymphopoietin (TSLP), overexpressed by skin keratinocytes, is the systemic driver of this bronchial hyper-responsiveness. As an AD-like model, we used mice with keratinocyte-specific deletion of RBP-j that sustained high systemic levels of TSLP. Antigen-induced allergic challenge to the lung airways of RBP-j-deficient animals resulted in a severe asthmatic phenotype not seen in similarly treated wild-type littermates. Elimination of TSLP signaling in these animals blocked the atopic march, demonstrating that high serum TSLP levels were required to sensitize the lung to allergic inflammation. Furthermore, we analyzed outbred K14-TSLP(tg) mice that maintained high systemic levels of TSLP without developing any skin pathology. Importantly, epidermal-derived TSLP was sufficient to trigger the atopic march, sensitizing the lung airways to inhaled allergens in the absence of epicutaneous sensitization. Based on these findings, we propose that in addition to early treatment of the primary skin-barrier defects, selective inhibition of systemic TSLP may be the key to blocking the development of asthma in AD patients. PMID- 19557149 TI - Implementation of guidelines for the management of arterial hypertension. The impulsion study. AB - This study assessed the effects of a pilot best practice implementation enhancement program on the control of hypertension. We enrolled 697 consecutive known hypertensive patients with other vascular risk factors but free from overt vascular disease. There was no "control" group because it was considered unethical to deprive high-risk patients from "best medical treatment". Following a baseline visit, previously trained physicians aimed to improve adherence to lifestyle measures and drug treatment for hypertension and other vascular risk factors. Both at baseline and at study completion (after 6 months), a 1-page form was completed showing if patients achieved treatment targets. If not, the reasons why were recorded. This program enhanced compliance with lifestyle measures and increased the use of evidence-based medication. There was a substantial increase in the number of patients who achieved treatment targets for blood pressure (p<0.0001) and other vascular risk factors. In non-diabetic patients (n=585), estimated vascular risk (PROCAM risk engine) was significantly reduced by 41% (p<0.0001). There was also a 12% reduction in vascular risk according to the Framingham risk engine but this did not achieve significance (p=0.07). In conclusion, this is the first study to increase adherence to multiple interventions in hypertensive patients on an outpatient basis, both in primary care and teaching hospitals. Simple, relatively low cost measures (e.g. educating physicians and patients, distributing printed guidelines/brochures and completing a 1-page form) motivated both physicians and patients to achieve multiple treatment goals. Further work is needed to establish if the improvement observed is sustained. [ClinicalTrials.gov NCT00416611]. PMID- 19557150 TI - The incremental value of troponin biomarkers in risk stratification of acute coronary syndromes: is the relationship multiplicative? AB - STRUCTURED ABSTRACT: OBJECTIVE: To emphasize the importance of troponin in the context of a new score for risk stratifying acute coronary syndromes (ACS) patients. Although troponins have powerful prognostic value, current ACS scores do not fully capitalize this prognostic ability. Here, we weigh troponin status in a multiplicative manner to develop the TRACS score from previously published Rush score risk factors (RRF). METHODS: 2,866 ACS patients (46.7% troponin positive) from 9 centers comprising the TRACS registry, were randomly split into derivation (n=1,422) and validation (n=1,444) cohorts. In the derivation sample, RRF sum was multiplied by 3 if troponins were positive to yield the TRACS score, which was grouped into five categories of 0-2, 3-5, 6-8, 9-11, 12-15 (multiples of 3). Predictive performance of this score to predict hospital death was ascertained in the validation sample. RESULTS: The TRACS score had ROC AUC of 0.71 in the validation cohort. Logistic regression, Kaplan-Meier analysis, likelihood-ratio and Bayesian Information Criterion (BIC) test indicated that weighing troponin status with 3 in the TRACS score improved the prediction of mortality. Hosmer-Lemeshow test indicated sound model fit. CONCLUSIONS: We demonstrate that weighing troponin as a multiple of 3 yields robust prognostication of hospital mortality in ACS patients, when used in the context of the TRACS score. PMID- 19557151 TI - In Vitro evaluation of Enterococcus faecalis adhesion on various endodontic medicaments. AB - E. faecalis in endodontic infection represents a biofilm type of disease, which explains the bacteria's resistance to various antimicrobial compounds and the subsequent failure after endodontic treatment. The purpose of this study was to compare antimicrobial activities and bacteria kinetic adhesion in vitro for three endodontic medicaments with a clinical isolate of E. faecalis. We devised a shake culture which contained the following intracanalar preparations: CPD, Endoidrox (EIX), PulpCanalSealer (PCS); these were immersed in a liquid culture medium inoculated with the microorganism. The shake system velocity was able to prevent non-specific bacteria adhesion and simulated the salivary flow. Specimens were collected daily (from both the medium and medicaments) for 10 days; the viable cells were counted by plate count, while the adhesion index AI degrees [E. faecalis fg DNA] /mm2 was evaluated in the pastes after DNA extraction, by quantitative real time PCR for the 16S rRNA gene. A partial growth inhibition, during the first 24 hours, was observed in the liquid medium and on the medicaments for EIX and subsequently for CPD (six logs). EIX showed the lowest adhesion coefficient (5*102 [fg DNA]/mm2) for nine days and was similar to the control. PCS showed no antimicrobial/antibiofilm properties. This showed that "calcium oxide" base compounds could be active against biofilm progression and at least in the short term (2-4 days) on E. faecalis cells growing in planktonic cultures. PMID- 19557152 TI - Acid resistance of the enamel in primary second molars from children with down syndrome and cerebral palsy. AB - OBJECTIVES: This study was carried out to evaluate the extent of differences in mineralization of inner and outer enamel of the lower primary second molars of children with Down syndrome (DS) and Cerebral Palsy (CP) as revealed by acid treatment of exfoliated teeth. The results were compared to those obtained from a control group of healthy children. METHODS: The sample included 4 mandibular second molars from each group. On each tooth, a thin section was cut, bisecting the mesial cusps. The analysis was carried out on the mesio-buccal cusps. Atomic force microscopy (AMF) was used to analyze the morphological structure of the dental enamel after 10 sec of 0.1 mol% citric acid treatment. The measurements were performed on 3 points in the enamel close to the outer surface and 3 points in the enamel close to the dentin. The differences between groups were analyzed using Mann Whitney tests. RESULTS: In controls and CP teeth the outer enamel was more resistant to etching than the inner enamel. In DS teeth both outer and inner enamel showed similar results for all parameters. Between group comparisons showed that roughness values were significantly higher (P<0.01) in DS teeth than in either controls or CP teeth. No significant differences were found between CP and control teeth. CONCLUSIONS: The higher values obtained for DS enamel reflect increased solubility of the enamel to acid relative to controls and CP teeth together with irregularity of the organic matrix. The practical importance of the results is that DS primary molars needs reduced etching time when prepared for pit and fissure sealants or composite/compomer restorations. PMID- 19557153 TI - Structural and ultra-structural features of the first mandibular molars of young rats submitted to pre and postnatal protein deficiencies. AB - The effects of protein malnutrition, both in utero and prior to weaning, on formation of the first mandibular molars were evaluated by phase-contrast and electron microscopy in rats. The nourished group (GI) received a diet that included 20% casein, while the malnourished group (GII) received 5% casein. The first mandibular molars from GII exhibited low density of cells and odontoblasts, which lacked regular organization compared with molars from GI. In addition, a difference in collagen type was observed between the groups, with a prevalence of Type III collagen fibers detected in the dentin, periodontal ligament, and alveolar bone of GII, and a prevalence of Type I collagen fibers in GI. Finally, examination of surface area in molar sagittal sections indicated 30% less dentin in GII, compared with GI. Our results suggest that structural and ultra structural features of the dentin-pulp complex and periodontal components of rat molars are affected by protein deficiency. PMID- 19557154 TI - Efflux in fungi: la piece de resistance. AB - Pathogens must be able to overcome both host defenses and antimicrobial treatment in order to successfully infect and maintain colonization of the host. One way fungi accomplish this feat and overcome intercellular toxin accumulation is efflux pumps, in particular ATP-binding cassette transporters and transporters of the major facilitator superfamily. Members of these two superfamilies remove many toxic compounds by coupling transport with ATP hydrolysis or a proton gradient, respectively. Fungal genomes encode a plethora of members of these families of transporters compared to other organisms. In this review we discuss the role these two fungal superfamilies of transporters play in virulence and resistance to antifungal agents. These efflux transporters are responsible not only for export of compounds involved in pathogenesis such as secondary metabolites, but also export of host-derived antimicrobial compounds. In addition, we examine the current knowledge of these transporters in resistance of pathogens to clinically relevant antifungal agents. PMID- 19557155 TI - Primary Structure Revision and Active Site Mapping of E. Coli Isoleucyl-tRNA Synthetase by Means of Maldi Mass Spectrometry. AB - The correct amino acid sequence of E. coli isoleucyl-tRNA synthetase (IleRS) was established by means of peptide mapping by MALDI mass spectrometry, using a set of four endoproteases (trypsin, LysC, AspN and GluC). Thereafter, the active site of IleRS was mapped by affinity labeling with reactive analogs of the substrates. For the ATP binding site, the affinity labeling reagent was pyridoxal 5' diphospho-5'-adenosine (ADP-PL), whereas periodate-oxidized tRNA(Ile), the 2',3' dialdehyde derivative of tRNA(Ile) was used to label the binding site for the 3' end of tRNA on the synthetase. Incubation of either reagent with IleRS resulted in a rapid loss of both the tRNA(Ile) aminoacylation and isoleucinedependent isotopic ATP-PPi exchange activities. The stoichiometries of IleRS labeling by ADP-PL or tRNA(Ile)ox corresponded to 1 mol of reagent incorporated per mol of enzyme. Altogether, the oxidized 3'-end of tRNA(Ile) and the pyridoxal moiety of the ATP analog ADP-PL react with the lysyl residues 601 and 604 of the consensus sequence (601)KMSKS(605). Identification of the binding site for L-isoleucine or for non cognate amino acids on E. coli IleRS was achieved by qualitative comparative labeling of the synthetase with bromomethyl ketone derivatives of L isoleucine (IBMK) or of the non-cognate amino acids valine (VBMK), phenylalanine (FBMK) and norleucine (NleBMK). Labeling of the enzyme with IBMK resulted in a complete loss of isoleucine-dependent isotopic [(32)P]PPi-ATP exchange activity. VBMK, NleBMK and FBMK were also capable of abolishing the activity of IleRS, FBMK being the less efficient in inactivating the synthetase. Analysis by MALDI mass spectrometry designated cysteines-462 and -718 as the target residues of the substrate analog IBMK on E. coli IleRS, whereas VBMK, NleBMK and FBMK labeled in common His-394, His-478 and Cys-718. In addition, VBMK and NleBMK, which are chemically similar to IBMK, were found covalently bound to Cys-462, and VBMK was specifically attached to His-332 (or His-337) of the synthetase. The amino acid residues labeled by the substrate analogs are mainly distributed between three regions in the primary structure of E. coli IleRS: these are segments [325-394], [451-479] and [591-604]. In the 3-D structures of IleRS from T. thermophilus and S. aureus, the [325-394] stretch is part of the editing domain, while fragments [451-479] and [591-604] representing the isoleucine binding domain and the dinucleotide (or Rossmann) fold domain, respectively, are located in the catalytic core. His-332 of E. coli IleRS, that is strictly conserved among all the available IleRS sequences is located in the editing active site of the synthetase. It is proposed that His-332 of E. coli IleRS participates directly in hydrolysis, or helps to deprotonate the hydroxyl group of threonine at the hydrolytic site. PMID- 19557156 TI - Stage-specific inhibition of MHC class I presentation by the Epstein-Barr virus BNLF2a protein during virus lytic cycle. AB - The gamma-herpesvirus Epstein-Barr virus (EBV) persists for life in infected individuals despite the presence of a strong immune response. During the lytic cycle of EBV many viral proteins are expressed, potentially allowing virally infected cells to be recognized and eliminated by CD8+ T cells. We have recently identified an immune evasion protein encoded by EBV, BNLF2a, which is expressed in early phase lytic replication and inhibits peptide- and ATP-binding functions of the transporter associated with antigen processing. Ectopic expression of BNLF2a causes decreased surface MHC class I expression and inhibits the presentation of indicator antigens to CD8+ T cells. Here we sought to examine the influence of BNLF2a when expressed naturally during EBV lytic replication. We generated a BNLF2a-deleted recombinant EBV (DeltaBNLF2a) and compared the ability of DeltaBNLF2a and wild-type EBV-transformed B cell lines to be recognized by CD8+ T cell clones specific for EBV-encoded immediate early, early and late lytic antigens. Epitopes derived from immediate early and early expressed proteins were better recognized when presented by DeltaBNLF2a transformed cells compared to wild-type virus transformants. However, recognition of late antigens by CD8+ T cells remained equally poor when presented by both wild-type and DeltaBNLF2a cell targets. Analysis of BNLF2a and target protein expression kinetics showed that although BNLF2a is expressed during early phase replication, it is expressed at a time when there is an upregulation of immediate early proteins and initiation of early protein synthesis. Interestingly, BNLF2a protein expression was found to be lost by late lytic cycle yet DeltaBNLF2a-transformed cells in late stage replication downregulated surface MHC class I to a similar extent as wild-type EBV-transformed cells. These data show that BNLF2a-mediated expression is stage specific, affecting presentation of immediate early and early proteins, and that other evasion mechanisms operate later in the lytic cycle. PMID- 19557157 TI - Epigenetic regulation of HIV-1 latency by cytosine methylation. AB - Human immunodeficiency virus type 1 (HIV-1) persists in a latent state within resting CD4+ T cells of infected persons treated with highly active antiretroviral therapy (HAART). This reservoir must be eliminated for the clearance of infection. Using a cDNA library screen, we have identified methyl CpG binding domain protein 2 (MBD2) as a regulator of HIV-1 latency. Two CpG islands flank the HIV-1 transcription start site and are methylated in latently infected Jurkat cells and primary CD4+ T cells. MBD2 and histone deacetylase 2 (HDAC2) are found at one of these CpG islands during latency. Inhibition of cytosine methylation with 5-aza-2'deoxycytidine (aza-CdR) abrogates recruitment of MBD2 and HDAC2. Furthermore, aza-CdR potently synergizes with the NF-kappaB activators prostratin or TNF-alpha to reactivate latent HIV-1. These observations confirm that cytosine methylation and MBD2 are epigenetic regulators of HIV-1 latency. Clearance of HIV-1 from infected persons may be enhanced by inclusion of DNA methylation inhibitors, such as aza-CdR, and NF-kappaB activators into current antiviral therapies. PMID- 19557158 TI - The structure of a biologically active influenza virus ribonucleoprotein complex. AB - The influenza viruses contain a segmented, single-stranded RNA genome of negative polarity. Each RNA segment is encapsidated by the nucleoprotein and the polymerase complex into ribonucleoprotein particles (RNPs), which are responsible for virus transcription and replication. Despite their importance, information about the structure of these RNPs is scarce. We have determined the three dimensional structure of a biologically active recombinant RNP by cryo-electron microscopy. The structure shows a nonameric nucleoprotein ring (at 12 Angstrom resolution) with two monomers connected to the polymerase complex (at 18 Angstrom resolution). Docking the atomic structures of the nucleoprotein and polymerase domains, as well as mutational analyses, has allowed us to define the interactions between the functional elements of the RNP and to propose the location of the viral RNA. Our results provide the first model for a functional negative-stranded RNA virus ribonucleoprotein complex. The structure reported here will serve as a framework to generate a quasi-atomic model of the molecular machine responsible for viral RNA synthesis and to test new models for virus RNA replication and transcription. PMID- 19557159 TI - Epstein-barr virus latency in B cells leads to epigenetic repression and CpG methylation of the tumour suppressor gene Bim. AB - In human B cells infected with Epstein-Barr virus (EBV), latency-associated virus gene products inhibit expression of the pro-apoptotic Bcl-2-family member Bim and enhance cell survival. This involves the activities of the EBV nuclear proteins EBNA3A and EBNA3C and appears to be predominantly directed at regulating Bim mRNA synthesis, although post-transcriptional regulation of Bim has been reported. Here we show that protein and RNA stability make little or no contribution to the EBV-associated repression of Bim in latently infected B cells. However, treatment of cells with inhibitors of histone deacetylase (HDAC) and DNA methyltransferase (DNMT) enzymes indicated that epigenetic mechanisms are involved in the down regulation of Bim. This was initially confirmed by chromatin immunoprecipitation analysis of histone acetylation levels on the Bim promoter. Consistent with this, methylation-specific PCR (MSP) and bisulphite sequencing of regions within the large CpG island located at the 5' end of Bim revealed significant methylation of CpG dinucleotides in all EBV-positive, but not EBV-negative B cells examined. Genomic DNA samples exhibiting methylation of the Bim promoter included extracts from a series of explanted EBV-positive Burkitt's lymphoma (BL) biopsies. Subsequent analyses of the histone modification H3K27-Me3 (trimethylation of histone H3 lysine 27) and CpG methylation at loci throughout the Bim promoter suggest that in EBV-positive B cells repression of Bim is initially associated with this repressive epigenetic histone mark gradually followed by DNA methylation at CpG dinucleotides. We conclude that latent EBV initiates a chain of events that leads to epigenetic repression of the tumour suppressor gene Bim in infected B cells and their progeny. This reprogramming of B cells could have important implications for our understanding of EBV persistence and the pathogenesis of EBV-associated disease, in particular BL. PMID- 19557160 TI - Maximum-likelihood model averaging to profile clustering of site types across discrete linear sequences. AB - A major analytical challenge in computational biology is the detection and description of clusters of specified site types, such as polymorphic or substituted sites within DNA or protein sequences. Progress has been stymied by a lack of suitable methods to detect clusters and to estimate the extent of clustering in discrete linear sequences, particularly when there is no a priori specification of cluster size or cluster count. Here we derive and demonstrate a maximum likelihood method of hierarchical clustering. Our method incorporates a tripartite divide-and-conquer strategy that models sequence heterogeneity, delineates clusters, and yields a profile of the level of clustering associated with each site. The clustering model may be evaluated via model selection using the Akaike Information Criterion, the corrected Akaike Information Criterion, and the Bayesian Information Criterion. Furthermore, model averaging using weighted model likelihoods may be applied to incorporate model uncertainty into the profile of heterogeneity across sites. We evaluated our method by examining its performance on a number of simulated datasets as well as on empirical polymorphism data from diverse natural alleles of the Drosophila alcohol dehydrogenase gene. Our method yielded greater power for the detection of clustered sites across a breadth of parameter ranges, and achieved better accuracy and precision of estimation of clusters, than did the existing empirical cumulative distribution function statistics. PMID- 19557162 TI - iNOS-producing inflammatory dendritic cells constitute the major infected cell type during the chronic Leishmania major infection phase of C57BL/6 resistant mice. AB - Leishmania major parasites reside and multiply in late endosomal compartments of host phagocytic cells. Immune control of Leishmania growth absolutely requires expression of inducible Nitric Oxide Synthase (iNOS/NOS2) and subsequent production of NO. Here, we show that CD11b+ CD11c+ Ly-6C+ MHC-II+ cells are the main iNOS-producing cells in the footpad lesion and in the draining lymph node of Leishmania major-infected C57BL/6 mice. These cells are phenotypically similar to iNOS-producing inflammatory DC (iNOS-DC) observed in the mouse models of Listeria monocytogenes and Brucella melitensis infection. The use of DsRed-expressing parasites demonstrated that these iNOS-producing cells are the major infected population in the lesions and the draining lymph nodes. Analysis of various genetically deficient mouse strains revealed the requirement of CCR2 expression for the recruitment of iNOS-DC in the draining lymph nodes, whereas their activation is strongly dependent on CD40, IL-12, IFN-gamma and MyD88 molecules with a partial contribution of TNF-alpha and TLR9. In contrast, STAT-6 deficiency enhanced iNOS-DC recruitment and activation in susceptible BALB/c mice, demonstrating a key role for IL-4 and IL-13 as negative regulators. Taken together, our results suggest that iNOS-DC represent a major class of Th1 regulated effector cell population and constitute the most frequent infected cell type during chronic Leishmania major infection phase of C57BL/6 resistant mice. PMID- 19557161 TI - Genome-wide association scan meta-analysis identifies three Loci influencing adiposity and fat distribution. AB - To identify genetic loci influencing central obesity and fat distribution, we performed a meta-analysis of 16 genome-wide association studies (GWAS, N = 38,580) informative for adult waist circumference (WC) and waist-hip ratio (WHR). We selected 26 SNPs for follow-up, for which the evidence of association with measures of central adiposity (WC and/or WHR) was strong and disproportionate to that for overall adiposity or height. Follow-up studies in a maximum of 70,689 individuals identified two loci strongly associated with measures of central adiposity; these map near TFAP2B (WC, P = 1.9x10(-11)) and MSRA (WC, P = 8.9x10( 9)). A third locus, near LYPLAL1, was associated with WHR in women only (P = 2.6x10(-8)). The variants near TFAP2B appear to influence central adiposity through an effect on overall obesity/fat-mass, whereas LYPLAL1 displays a strong female-only association with fat distribution. By focusing on anthropometric measures of central obesity and fat distribution, we have identified three loci implicated in the regulation of human adiposity. PMID- 19557163 TI - The glyceraldehyde-3-phosphate dehydrogenase and the small GTPase Rab 2 are crucial for Brucella replication. AB - The intracellular pathogen Brucella abortus survives and replicates inside host cells within an endoplasmic reticulum (ER)-derived replicative organelle named the "Brucella-containing vacuole" (BCV). Here, we developed a subcellular fractionation method to isolate BCVs and characterize for the first time the protein composition of its replicative niche. After identification of BCV membrane proteins by 2 dimensional (2D) gel electrophoresis and mass spectrometry, we focused on two eukaryotic proteins: the glyceraldehyde-3 phosphate dehydrogenase (GAPDH) and the small GTPase Rab 2 recruited to the vacuolar membrane of Brucella. These proteins were previously described to localize on vesicular and tubular clusters (VTC) and to regulate the VTC membrane traffic between the endoplasmic reticulum (ER) and the Golgi. Inhibition of either GAPDH or Rab 2 expression by small interfering RNA strongly inhibited B. abortus replication. Consistent with this result, inhibition of other partners of GAPDH and Rab 2, such as COPI and PKC iota, reduced B. abortus replication. Furthermore, blockage of Rab 2 GTPase in a GDP-locked form also inhibited B. abortus replication. Bacteria did not fuse with the ER and instead remained in lysosomal-associated membrane vacuoles. These results reveal an essential role for GAPDH and the small GTPase Rab 2 in B. abortus virulence within host cells. PMID- 19557164 TI - Assessing the impact of transgenerational epigenetic variation on complex traits. AB - Loss or gain of DNA methylation can affect gene expression and is sometimes transmitted across generations. Such epigenetic alterations are thus a possible source of heritable phenotypic variation in the absence of DNA sequence change. However, attempts to assess the prevalence of stable epigenetic variation in natural and experimental populations and to quantify its impact on complex traits have been hampered by the confounding effects of DNA sequence polymorphisms. To overcome this problem as much as possible, two parents with little DNA sequence differences, but contrasting DNA methylation profiles, were used to derive a panel of epigenetic Recombinant Inbred Lines (epiRILs) in the reference plant Arabidopsis thaliana. The epiRILs showed variation and high heritability for flowering time and plant height ( approximately 30%), as well as stable inheritance of multiple parental DNA methylation variants (epialleles) over at least eight generations. These findings provide a first rationale to identify epiallelic variants that contribute to heritable variation in complex traits using linkage or association studies. More generally, the demonstration that numerous epialleles across the genome can be stable over many generations in the absence of selection or extensive DNA sequence variation highlights the need to integrate epigenetic information into population genetics studies. PMID- 19557166 TI - Pandemic vaccine preparedness--have we left something behind? PMID- 19557165 TI - Ebola Zaire virus blocks type I interferon production by exploiting the host SUMO modification machinery. AB - Ebola Zaire virus is highly pathogenic for humans, with case fatality rates approaching 90% in large outbreaks in Africa. The virus replicates in macrophages and dendritic cells (DCs), suppressing production of type I interferons (IFNs) while inducing the release of large quantities of proinflammatory cytokines. Although the viral VP35 protein has been shown to inhibit IFN responses, the mechanism by which it blocks IFN production has not been fully elucidated. We expressed VP35 from a mouse-adapted variant of Ebola Zaire virus in murine DCs by retroviral gene transfer, and tested for IFN transcription upon Newcastle Disease virus (NDV) infection and toll-like receptor signaling. We found that VP35 inhibited IFN transcription in DCs following these stimuli by disabling the activity of IRF7, a transcription factor required for IFN transcription. By yeast two-hybrid screens and coimmunoprecipitation assays, we found that VP35 interacted with IRF7, Ubc9 and PIAS1. The latter two are the host SUMO E2 enzyme and E3 ligase, respectively. VP35, while not itself a SUMO ligase, increased PIAS1-mediated SUMOylation of IRF7, and repressed Ifn transcription. In contrast, VP35 did not interfere with the activation of NF-kappaB, which is required for induction of many proinflammatory cytokines. Our findings indicate that Ebola Zaire virus exploits the cellular SUMOylation machinery for its advantage and help to explain how the virus overcomes host innate defenses, causing rapidly overwhelming infection to produce a syndrome resembling fulminant septic shock. PMID- 19557167 TI - Differential effects of early weaning for HIV-free survival of children born to HIV-infected mothers by severity of maternal disease. AB - BACKGROUND: We previously reported no benefit of early weaning for HIV-free survival of children born to HIV-infected mothers in intent-to-treat analyses. Since early weaning was poorly accepted, we conducted a secondary analysis to investigate whether beneficial effects may have been hidden. METHODS: 958 HIV infected women in Lusaka, Zambia, were randomized to abrupt weaning at 4 months (intervention) or to continued breastfeeding (control). Children were followed to 24 months with regular HIV PCR tests and examinations to determine HIV infection or death. Detailed behavioral data were collected on when all breastfeeding ended. Most participants were recruited before antiretroviral treatment (ART) became available. We compared outcomes among mother-child pairs who weaned earlier or later than intended by study design adjusting for potential confounders. RESULTS: Of infants alive, uninfected and still breastfeeding at 4 months in the intervention group, 16.1% who weaned as instructed acquired HIV or died by 24 months compared to 16.0% who did not comply (p = 0.98). Children of women with less severe disease during pregnancy (not eligible for ART) had worse outcomes if their mothers weaned as instructed (RH = 2.60 95% CI: 1.06-6.36) compared to those who continued breastfeeding. Conversely, children of mothers with more severe disease (eligible for ART but did not receive it) who weaned early had better outcomes (p-value interaction = 0.002). In the control group, weaning before 15 months was associated with 3.94-fold (95% CI: 1.65-9.39) increase in HIV infection or death among infants of mothers with less severe disease. CONCLUSION: Incomplete adherence did not mask a benefit of early weaning. On the contrary, for women with less severe disease, early weaning was harmful and continued breastfeeding resulted in better outcomes. For women with more advanced disease, ART should be given during pregnancy for maternal health and to reduce transmission, including through breastfeeding. TRIAL REGISTRATION: (ClinicalTrials.gov) NCT00310726. PMID- 19557168 TI - Concurrent growth rate and transcript analyses reveal essential gene stringency in Escherichia coli. AB - BACKGROUND: Genes essential for bacterial growth are of particular scientific interest. Many putative essential genes have been identified or predicted in several species, however, little is known about gene expression requirement stringency, which may be an important aspect of bacterial physiology and likely a determining factor in drug target development. METHODOLOGY/PRINCIPAL FINDINGS: Working from the premise that essential genes differ in absolute requirement for growth, we describe silencing of putative essential genes in E. coli to obtain a titration of declining growth rates and transcript levels by using antisense peptide nucleic acids (PNA) and expressed antisense RNA. The relationship between mRNA decline and growth rate decline reflects the degree of essentiality, or stringency, of an essential gene, which is here defined by the minimum transcript level for a 50% reduction in growth rate (MTL(50)). When applied to four growth essential genes, both RNA silencing methods resulted in MTL(50) values that reveal acpP as the most stringently required of the four genes examined, with ftsZ the next most stringently required. The established antibacterial targets murA and fabI were less stringently required. CONCLUSIONS: RNA silencing can reveal stringent requirements for gene expression with respect to growth. This method may be used to validate existing essential genes and to quantify drug target requirement. PMID- 19557169 TI - Bacterial antigen expression is an important component in inducing an immune response to orally administered Salmonella-delivered DNA vaccines. AB - BACKGROUND: The use of Salmonella to deliver heterologous antigens from DNA vaccines is a well-accepted extension of the success of oral Salmonella vaccines in animal models. Attenuated S. typhimurium and S. typhi strains are safe and efficacious, and their use to deliver DNA vaccines combines the advantages of both vaccine approaches, while complementing the limitations of each technology. An important aspect of the basic biology of the Salmonella/DNA vaccine platform is the relative contributions of prokaryotic and eukaryotic expression in production of the vaccine antigen. Gene expression in DNA vaccines is commonly under the control of the eukaryotic cytomegalovirus (CMV) promoter. The aim of this study was to identify and disable putative bacterial promoters within the CMV promoter and evaluate the immunogenicity of the resulting DNA vaccine delivered orally by S. typhimurium. METHODOLOGY/PRINCIPAL FINDINGS: The results reported here clearly demonstrate the presence of bacterial promoters within the CMV promoter. These promoters have homology to the bacterial consensus sequence and functional activity. To disable prokaryotic expression from the CMV promoter a series of genetic manipulations were performed to remove the two major bacterial promoters and add a bacteria transcription terminator downstream of the CMV promoter. S. typhimurium was used to immunise BALB/c mice orally with a DNA vaccine encoding the C-fragment of tetanus toxin (TT) under control of the original or the modified CMV promoter. Although both promoters functioned equally well in eukaryotic cells, as indicated by equivalent immune responses following intramuscular delivery, only the original CMV promoter was able to induce an anti TT specific response following oral delivery by S. typhimurium. CONCLUSIONS: These findings suggest that prokaryotic expression of the antigen and co-delivery of this protein by Salmonella are at least partially responsible for the successful oral delivery of C-fragment DNA vaccines containing the CMV promoter by S. typhimurium. PMID- 19557170 TI - Quantification of food intake in Drosophila. AB - Measurement of food intake in the fruit fly Drosophila melanogaster is often necessary for studies of behaviour, nutrition and drug administration. There is no reliable and agreed method for measuring food intake of flies in undisturbed, steady state, and normal culture conditions. We report such a method, based on measurement of feeding frequency by proboscis-extension, validated by short-term measurements of food dye intake. We used the method to demonstrate that (a) female flies feed more frequently than males, (b) flies feed more often when housed in larger groups and (c) fly feeding varies at different times of the day. We also show that alterations in food intake are not induced by dietary restriction or by a null mutation of the fly insulin receptor substrate chico. In contrast, mutation of takeout increases food intake by increasing feeding frequency while mutation of ovo(D) increases food intake by increasing the volume of food consumed per proboscis-extension. This approach provides a practical and reliable method for quantification of food intake in Drosophila under normal, undisturbed culture conditions. PMID- 19557171 TI - Competition triggers plasmid-mediated enhancement of substrate utilisation in Pseudomonas putida. AB - Competition between species plays a central role in the activity and structure of communities. Stable co-existence of diverse organisms in communities is thought to be fostered by individual tradeoffs and optimization of competitive strategies along resource gradients. Outside the laboratory, microbes exist as multispecies consortia, continuously interacting with one another and the environment. Survival and proliferation of a particular species is governed by its competitive fitness. Therefore, bacteria must be able to continuously sense their immediate environs for presence of competitors and prevailing conditions. Here we present results of our investigations on a novel competition sensing mechanism in the rhizosphere-inhabiting Pseudomonas putida KT2440, harbouring gfpmut3b-modified Kan(R) TOL plasmid. We monitored benzyl alcohol (BA) degradation rate, along with GFP expression profiling in mono species and dual species cultures. Interestingly, enhanced plasmid expression (monitored using GFP expression) and consequent BA degradation were observed in dual species consortia, irrespective of whether the competitor was a BA degrader (Pseudomonas aeruginosa) or a non degrader (E. coli). Attempts at elucidation of the mechanistic aspects of induction indicated the role of physical interaction, but not of any diffusible compounds emanating from the competitors. This contention is supported by the observation that greater induction took place in presence of increasing number of competitors. Inert microspheres mimicking competitor cell size and concentration did not elicit any significant induction, further suggesting the role of physical cell-cell interaction. Furthermore, it was also established that cell wall compromised competitor had minimal induction capability. We conclude that P. putida harbouring pWW0 experience a competitive stress when grown as dual-species consortium, irrespective of the counterpart being BA degrader or not. The immediate effect of this stress is a marked increase in expression of TOL, leading to rapid utilization of the available carbon source and massive increase in its population density. The plausible mechanisms behind the phenomenon are hypothesised and practical implications are indicated and discussed. PMID- 19557172 TI - The Wnt/beta-catenin pathway interacts differentially with PTHrP signaling to control chondrocyte hypertrophy and final maturation. AB - Sequential proliferation, hypertrophy and maturation of chondrocytes are required for proper endochondral bone development and tightly regulated by cell signaling. The canonical Wnt signaling pathway acts through beta-catenin to promote chondrocyte hypertrophy whereas PTHrP signaling inhibits it by holding chondrocytes in proliferating states. Here we show by genetic approaches that chondrocyte hypertrophy and final maturation are two distinct developmental processes that are differentially regulated by Wnt/beta-catenin and PTHrP signaling. Wnt/beta-catenin signaling regulates initiation of chondrocyte hypertrophy by inhibiting PTHrP signaling activity, but it does not regulate PTHrP expression. In addition, Wnt/beta-catenin signaling regulates chondrocyte hypertrophy in a non-cell autonomous manner and Gdf5/Bmp signaling may be one of the downstream pathways. Furthermore, Wnt/beta-catenin signaling also controls final maturation of hypertrophic chondrocytes, but such regulation is PTHrP signaling-independent. PMID- 19557173 TI - Interaction with LC8 is required for Pak1 nuclear import and is indispensable for zebrafish development. AB - Pak1 (p21 activated kinase 1) is a serine/threonine kinase implicated in regulation of cell motility and survival and in malignant transformation of mammary epithelial cells. In addition, the dynein light chain, LC8, has been described to cooperate with Pak1 in malignant transformation of breast cancer cells. Pak1 itself may aid breast cancer development by phosphorylating nuclear proteins, including estrogen receptor alpha. Recently, we showed that the LC8 binding site on Pak1 is adjacent to the nuclear localization sequence (NLS) required for Pak1 nuclear import. Here, we demonstrate that the LC8-Pak1 interaction is necessary for epidermal growth factor (EGF)-induced nuclear import of Pak1 in MCF-7 cells, and that this event is contingent upon LC8-mediated Pak1 dimerization. In contrast, Pak2, which lacks an LC8 binding site but contains a nuclear localization sequence identical to that in Pak1, remains cytoplasmic upon EGF stimulation of MCF-7 cells. Furthermore, we show that severe developmental defects in zebrafish embryos caused by morpholino injections targeting Pak are partially rescued by co-injection of wild-type human Pak1, but not by co injection of mutant Pak1 mRNA disrupting either the LC8 binding or the NLS site. Collectively, these results suggest that LC8 facilitates nuclear import of Pak1 and that this function is indispensable during vertebrate development. PMID- 19557174 TI - Increased microRNA activity in human cancers. AB - MicroRNAs (miRNAs) are small regulatory RNAs that act by blocking the translation and increasing the degradation of target transcripts. MiRNAs play a critical role in many biological processes including development and differentiation and many studies have shown that major changes in miRNA levels occur in cancer. Since miRNAs degrade target messages, we used this property to develop a novel computational method aimed at determining the actual biological activity of miRNAs using variations in gene expression. Using the method described here, we quantified miRNA activity in papillary thyroid carcinoma and breast cancer, and found a strong and distinctive signal of increased global miRNA activity, embedded in the pertaining gene expression measurements. Interestingly, we found that in these two cancers, miRNA activity is globally increased, and is associated with a global downregulation of miRNA target genes. This downregulation of miRNA regulated genes is particularly noticeable for genes carrying multiple target sites for miRNAs. Among the miRNA-repressed genes, we found a significant enrichment of known tumor suppressors, thereby suggesting that the increased miRNA activity was indeed tumorigenic. PMID- 19557175 TI - No detectable maternal effects of elevated CO(2) on Arabidopsis thaliana over 15 generations. AB - Maternal environment has been demonstrated to produce considerable impact on offspring growth. However, few studies have been carried out to investigate multi generational maternal effects of elevated CO(2) on plant growth and development. Here we present the first report on the responses of plant reproductive, photosynthetic, and cellular characteristics to elevated CO(2) over 15 generations using Arabidopsis thaliana as a model system. We found that within an individual generation, elevated CO(2) significantly advanced plant flowering, increased photosynthetic rate, increased the size and number of starch grains per chloroplast, reduced stomatal density, stomatal conductance, and transpiration rate, and resulted in a higher reproductive mass. Elevated CO(2) did not significantly influence silique length and number of seeds per silique. Across 15 generations grown at elevated CO(2) concentrations, however, there were no significant differences in these traits. In addition, a reciprocal sowing experiment demonstrated that elevated CO(2) did not produce detectable maternal effects on the offspring after fifteen generations. Taken together, these results suggested that the maternal effects of elevated CO(2) failed to extend to the offspring due to the potential lack of genetic variation for CO(2) responsiveness, and future plants may not evolve specific adaptations to elevated CO(2) concentrations. PMID- 19557176 TI - The role of salivary and intestinal complement system inhibitors in the midgut protection of triatomines and mosquitoes. AB - Saliva of haematophagous arthropods contain biomolecules involved directly or indirectly with the haematophagy process, and among them are encountered some complement system inhibitors. The most obvious function for these inhibitors would be the protection of the midgut against injury by the complement. To investigate this hypothesis, Triatoma brasiliensis nymphs were forced to ingest human serum in conditions in which the protection of midgut by the inhibitors is bypassed. In these conditions, the anterior midgut epithelium was injured by the complement, causing cell death. Once some insects such as Aedes aegypti have no salivary inhibitors, we hypothesized the existence of intestinal inhibitors. The inhibitory activity was investigated in the intestine of A. aegypti as well as in the saliva and intestine of other three triatomine species (T. brasiliensis, T. infestans and Rhodnius prolixus) using an immunological method able to determine the level of deposition of some complement factors (C1q, C3b, or C4b) on the surface of complement activator molecules linked to microplates. This methodology permitted to identify which points along the activation phase of the complement cascade were inhibited. As expected, soluble contents of A. aegypti's intestine was capable to inhibit C3b deposition by the classical and alternative pathways. Saliva or soluble intestinal contents, obtained from triatomines were unable to inhibit C1q deposition by the classical pathway. C4b deposition by the classical pathway was inhibited by the intestinal contents from the three triatomines. On the other hand, only T. brasiliensis saliva inhibited C4b deposition. Both, saliva and intestinal contents from all triatomines were able to inhibit C3b deposition in the classical and alternative pathways. None of the material extracted from the intestinal cell membranes from the triatomines inhibited C3b deposition in the classical pathway. The existence of complement inhibitors may have important biological consequences which are discussed in detail. PMID- 19557177 TI - Tbx1 regulates the BMP-Smad1 pathway in a transcription independent manner. AB - Tbx1 is a T-box transcription factor implicated in DiGeorge syndrome. The molecular function of Tbx1 is unclear although it can transactivate reporters with T-box binding elements. We discovered that Tbx1 binds Smad1 and suppresses the Bmp4/Smad1 signaling. Tbx1 interferes with Smad1 to Smad4 binding, and a mutation of Tbx1 that abolishes transactivation, does not affect Smad1 binding nor does affect the ability to suppress Smad1 activity. In addition, a disease associated mutation of TBX1 that does not prevent transactivation, prevents the TBX1-SMAD1 interaction. Expression of Tbx1 in transgenic mice generates phenotypes similar to those associated with loss of a Bmp receptor. One phenotype could be rescued by transgenic Smad1 expression. Our data indicate that Tbx1 interferes with Bmp/Smad1 signaling and provide strong evidence that a T-box transcription factor has functions unrelated to transactivation. PMID- 19557178 TI - 3D model of lamprey estrogen receptor with estradiol and 15alpha-hydroxy estradiol. AB - BACKGROUND: Lamprey, basal vertebrate, is an important model system for understanding early events in vertebrate evolution. Lamprey contains orthologs of the estrogen receptor [ER], progesterone receptor and corticoid receptor. A perplexing property of lamprey is that 15alpha-hydroxy-steroids are active steroids. For example, 15alpha-hydroxy-estradiol [15alpha-OH-E2] is the estrogen, instead of estradiol [E2]. To investigate how 15alpha-OH-E2 binds lamprey ER, we constructed a 3D model of the lamprey ER with E2 and 15alpha-OH-E2. METHODOLOGY: We used the 3D structure of human ERalpha as a template to construct a 3D model of lamprey ER. E2 and 15alpha-OH-E2 were inserted into the 3D model of lamprey ER and 15alpha-OH-E2 was inserted into human ERalpha. Then the each steroid-protein complex was refined using Discover 3 from Insight II software. To determine if lamprey ER had some regions that were unique among vertebrate ERs, we used the ligand-binding domain of lamprey ER as a query for a BLAST search of GenBank. PRINCIPAL FINDINGS: Our 3D model of lamprey ER with 15alpha-OH-E2 shows that Sdelta on Met-409 can form a hydrogen bond with the 15alpha-hydroxyl on 15alpha OH-E2. In human ERalpha, the corresponding residue Ile-424 has a van der Waals contact with 15alpha-OH-E2. BLAST analysis of GenBank indicates that among vertebrate ERs, only lamprey ER contains a methionine at this position. Thus, the contact between Sdelta on Met-409 and 15alpha-OH-E2 is unique. Interestingly, BLAST finds that five New World monkeys and a sturgeon contain a valine instead of isoleucine. SIGNIFICANCE: In addition to shedding light on the structure of the ER in a basal vertebrate, our 3D model of lamprey ER should prove useful in virtual screening of chemical libraries to identify compounds for controlling reproduction in sea lamprey, an environmental pest in Lake Michigan. PMID- 19557179 TI - Alteration of gene expression signatures of cortical differentiation and wound response in lethal clear cell renal cell carcinomas. AB - Clear cell renal cell carcinoma (ccRCC) is the most common malignancy of the adult kidney and displays heterogeneity in clinical outcomes. Through comprehensive gene expression profiling, we have identified previously a set of transcripts that predict survival following nephrectomy independent of tumor stage, grade, and performance status. These transcripts, designated as the SPC (supervised principal components) gene set, show no apparent biological or genetic features that provide insight into renal carcinogenesis or tumor progression. We explored the relationship of this gene list to a set of genes expressed in different anatomical segments of the normal kidney including the cortex (cortex gene set) and the glomerulus (glomerulus gene set), and a gene set expressed after serum stimulation of quiescent fibroblasts (the core serum response or CSR gene set). Interestingly, the normal cortex, glomerulus (part of the normal renal cortex), and CSR gene sets captured more than 1/5 of the genes in the highly prognostic SPC gene set. Based on gene expression patterns alone, the SPC gene set could be used to sort samples from normal adult kidneys by the anatomical regions from which they were dissected. Tumors whose gene expression profiles most resembled the normal renal cortex or glomerulus showed better survival than those that did not, and those with expression features more similar to CSR showed poorer survival. While the cortex, glomerulus, and CSR signatures predicted survival independent of traditional clinical parameters, they were not independent of the SPC gene list. Our findings suggest that critical biological features of lethal ccRCC include loss of normal cortical differentiation and activation of programs associated with wound healing. PMID- 19557180 TI - Genetic profiling reveals cross-contamination and misidentification of 6 adenoid cystic carcinoma cell lines: ACC2, ACC3, ACCM, ACCNS, ACCS and CAC2. AB - Adenoid cystic carcinoma (ACC) is the second most common malignant neoplasm of the salivary glands. Most patients survive more than 5 years after surgery and postoperative radiation therapy. The 10 year survival rate, however, drops to 40%, due to locoregional recurrences and distant metastases. Improving long-term survival in ACC requires the development of more effective systemic therapies based on a better understanding of the biologic behavior of ACC. Much preclinical research in this field involves the use of cultured cells and, to date, several ACC cell lines have been established. Authentication of these cell lines, however, has not been reported. We performed DNA fingerprint analysis on six ACC cell lines using short tandem repeat (STR) examinations and found that all six cell lines had been contaminated with other cells. ACC2, ACC3, and ACCM were determined to be cervical cancer cells (HeLa cells), whereas the ACCS cell line was composed of T24 urinary bladder cancer cells. ACCNS and CAC2 cells were contaminated with cells derived from non-human mammalian species: the cells labeled ACCNS were mouse cells and the CAC2 cells were rat cells. These observations suggest that future studies using ACC cell lines should include cell line authentication to avoid the use of contaminated or non-human cells. PMID- 19557181 TI - Reduced attentional scope in cocaine polydrug users. AB - Cocaine is Europe's second preferred recreational drug after cannabis but very little is known about possible cognitive impairments in the upcoming type of recreational cocaine user (monthly consumption). We asked whether recreational use of cocaine impacts early attentional selection processes. Cocaine-free polydrug controls (n = 18) and cocaine polydrug users (n = 18) were matched on sex, age, alcohol consumption, and IQ (using the Raven's progressive matrices), and were tested by using the Global-Local task to measure the scope of attention. Cocaine polydrug users attended significantly more to local aspects of attended events, which fits with the idea that a reduced scope of attention may be associated with the perpetuation of the use of the drug. PMID- 19557182 TI - A novel flow cytometric high throughput assay for a systematic study on molecular mechanisms underlying T cell receptor-mediated integrin activation. AB - Lymphocyte function-associated antigen 1 (LFA-1), a member of beta2-integrin family, exerts multiple roles in host T cell immunity and has been identified as a useful drug-development target for inflammatory and autoimmune diseases. Applying the findings that primary resting T cells absorb nanometric membrane vesicles derived from antigen presenting cells (APC) via dual receptor/ligand interactions of T cell receptor (TCR) with cognate peptide-major histocompatibility complex (MHC) complex (pMHC) and LFA-1 with its ligand, intercellular adhesion molecule-1 (ICAM-1), and that signaling cascades triggered by TCR/pMHC interaction take a part in the vesicle-absorption, we established a cell-based high throughput assay for systematic investigation, via isolation of small molecules modulating the level of vesicle-absorption, of molecular mechanisms underlying the T cell absorption of APC-derived vesicles, i.e., structural basis of TCR/pMHC and LFA-1/ICAM-1 interactions and TCR-mediated LFA-1 activation. As primary T cells along with physiological ligands expressed in biological membrane are used and also individual cells in assay samples are analyzed by flow cytometry, results obtained using the assay system hold superior physiological and therapeutic relevance as well as statistical precision. PMID- 19557183 TI - Dynamic interaction between STLV-1 proviral load and T-cell response during chronic infection and after immunosuppression in non-human primates. AB - We used mandrills (Mandrillus sphinx) naturally infected with simian T-cell leukemia virus type 1 (STLV-1) as a model for evaluating the influence of natural STLV-1 infection on the dynamics and evolution of the immune system during chronic infection. Furthermore, in order to evaluate the role of the immune system in controlling the infection during latency, we induced immunosuppression in the infected monkeys. We first showed that the STLV-1 proviral load was higher in males than in females and increased significantly with the duration of infection: mandrills infected for 10-6 years had a significantly higher proviral load than those infected for 2-4 years. Curiously, this observation was associated with a clear reduction in CD4+ T-cell number with age. We also found that the percentage of CD4(+) T cells co-expressing the activation marker HLA-DR and the mean percentage of CD25(+) in CD4(+) and CD8(+) T cells were significantly higher in infected than in uninfected animals. Furthermore, the STLV-1 proviral load correlated positively with T-cell activation but not with the frequency of T cells secreting interferon gamma in response to Tax peptides. Lastly, we showed that, during immunosuppression in infected monkeys, the percentages of CD8(+) T cells expressing HLA-DR(+) and of CD4(+) T cells expressing the proliferation marker Ki67 decreased significantly, although the percentage of CD8(+) T cells expressing HLA-DR(+) and Ki67 increased significantly by the end of treatment. Interestingly, the proviral load increased significantly after immunosuppression in the monkey with the highest load. Our study demonstrates that mandrills naturally infected with STLV-1 could be a suitable model for studying the relations between host and virus. Further studies are needed to determine whether the different compartments of the immune response during infection induce the long latency by controlling viral replication over time. Such studies would provide important information for the development of immune-based therapeutic strategies. PMID- 19557184 TI - The need for centralization of computational biology resources. PMID- 19557185 TI - Uptake of the necrotic serpin in Drosophila melanogaster via the lipophorin receptor-1. AB - The humoral response to fungal and Gram-positive infections is regulated by the serpin-family inhibitor, Necrotic. Following immune-challenge, a proteolytic cascade is activated which signals through the Toll receptor. Toll activation results in a range of antibiotic peptides being synthesised in the fat-body and exported to the haemolymph. As with mammalian serpins, Necrotic turnover in Drosophila is rapid. This serpin is synthesised in the fat-body, but its site of degradation has been unclear. By "freezing" endocytosis with a temperature sensitive Dynamin mutation, we demonstrate that Necrotic is removed from the haemolymph in two groups of giant cells: the garland and pericardial athrocytes. Necrotic uptake responds rapidly to infection, being visibly increased after 30 mins and peaking at 6-8 hours. Co-localisation of anti-Nec with anti-AP50, Rab5, and Rab7 antibodies establishes that the serpin is processed through multi vesicular bodies and delivered to the lysosome, where it co-localises with the ubiquitin-binding protein, HRS. Nec does not co-localise with Rab11, indicating that the serpin is not re-exported from athrocytes. Instead, mutations which block late endosome/lysosome fusion (dor, hk, and car) cause accumulation of Necrotic-positive endosomes, even in the absence of infection. Knockdown of the 6 Drosophila orthologues of the mammalian LDL receptor family with dsRNA identifies LpR1 as an enhancer of the immune response. Uptake of Necrotic from the haemolymph is blocked by a chromosomal deletion of LpR1. In conclusion, we identify the cells and the receptor molecule responsible for the uptake and degradation of the Necrotic serpin in Drosophila melanogaster. The scavenging of serpin/proteinase complexes may be a critical step in the regulation of proteolytic cascades. PMID- 19557186 TI - Forging links between human mental retardation-associated CNVs and mouse gene knockout models. AB - Rare copy number variants (CNVs) are frequently associated with common neurological disorders such as mental retardation (MR; learning disability), autism, and schizophrenia. CNV screening in clinical practice is limited because pathological CNVs cannot be distinguished routinely from benign CNVs, and because genes underlying patients' phenotypes remain largely unknown. Here, we present a novel, statistically robust approach that forges links between 148 MR-associated CNVs and phenotypes from approximately 5,000 mouse gene knockout experiments. These CNVs were found to be significantly enriched in two classes of genes, those whose mouse orthologues, when disrupted, result in either abnormal axon or dopaminergic neuron morphologies. Additional enrichments highlighted correspondences between relevant mouse phenotypes and secondary presentations such as brain abnormality, cleft palate, and seizures. The strength of these phenotype enrichments (>100% increases) greatly exceeded molecular annotations (<30% increases) and allowed the identification of 78 genes that may contribute to MR and associated phenotypes. This study is the first to demonstrate how the power of mouse knockout data can be systematically exploited to better understand genetically heterogeneous neurological disorders. PMID- 19557187 TI - Telomere recombination accelerates cellular aging in Saccharomyces cerevisiae. AB - Telomeres are nucleoprotein structures located at the linear ends of eukaryotic chromosomes. Telomere integrity is required for cell proliferation and survival. Although the vast majority of eukaryotic species use telomerase as a primary means for telomere maintenance, a few species can use recombination or retrotransposon-mediated maintenance pathways. Since Saccharomyces cerevisiae can use both telomerase and recombination to replicate telomeres, budding yeast provides a useful system with which to examine the evolutionary advantages of telomerase and recombination in preserving an organism or cell under natural selection. In this study, we examined the life span in telomerase-null, post senescent type II survivors that have employed homologous recombination to replicate their telomeres. Type II recombination survivors stably maintained chromosomal integrity but exhibited a significantly reduced replicative life span. Normal patterns of cell morphology at the end of a replicative life span and aging-dependent sterility were observed in telomerase-null type II survivors, suggesting the type II survivors aged prematurely in a manner that is phenotypically consistent with that of wild-type senescent cells. The shortened life span of type II survivors was extended by calorie restriction or TOR1 deletion, but not by Fob1p inactivation or Sir2p over-expression. Intriguingly, rDNA recombination was decreased in type II survivors, indicating that the premature aging of type II survivors was not caused by an increase in extra chromosomal rDNA circle accumulation. Reintroduction of telomerase activity immediately restored the replicative life span of type II survivors despite their heterogeneous telomeres. These results suggest that telomere recombination accelerates cellular aging in telomerase-null type II survivors and that telomerase is likely a superior telomere maintenance pathway in sustaining yeast replicative life span. PMID- 19557188 TI - Evolution of mutational robustness in the yeast genome: a link to essential genes and meiotic recombination hotspots. AB - Deleterious mutations inevitably emerge in any evolutionary process and are speculated to decisively influence the structure of the genome. Meiosis, which is thought to play a major role in handling mutations on the population level, recombines chromosomes via non-randomly distributed hot spots for meiotic recombination. In many genomes, various types of genetic elements are distributed in patterns that are currently not well understood. In particular, important (essential) genes are arranged in clusters, which often cannot be explained by a functional relationship of the involved genes. Here we show by computer simulation that essential gene (EG) clustering provides a fitness benefit in handling deleterious mutations in sexual populations with variable levels of inbreeding and outbreeding. We find that recessive lethal mutations enforce a selective pressure towards clustered genome architectures. Our simulations correctly predict (i) the evolution of non-random distributions of meiotic crossovers, (ii) the genome-wide anti-correlation of meiotic crossovers and EG clustering, (iii) the evolution of EG enrichment in pericentromeric regions and (iv) the associated absence of meiotic crossovers (cold centromeres). Our results furthermore predict optimal crossover rates for yeast chromosomes, which match the experimentally determined rates. Using a Saccharomyces cerevisiae conditional mutator strain, we show that haploid lethal phenotypes result predominantly from mutation of single loci and generally do not impair mating, which leads to an accumulation of mutational load following meiosis and mating. We hypothesize that purging of deleterious mutations in essential genes constitutes an important factor driving meiotic crossover. Therefore, the increased robustness of populations to deleterious mutations, which arises from clustered genome architectures, may provide a significant selective force shaping crossover distribution. Our analysis reveals a new aspect of the evolution of genome architectures that complements insights about molecular constraints, such as the interference of pericentromeric crossovers with chromosome segregation. PMID- 19557189 TI - Identifying relationships among genomic disease regions: predicting genes at pathogenic SNP associations and rare deletions. AB - Translating a set of disease regions into insight about pathogenic mechanisms requires not only the ability to identify the key disease genes within them, but also the biological relationships among those key genes. Here we describe a statistical method, Gene Relationships Among Implicated Loci (GRAIL), that takes a list of disease regions and automatically assesses the degree of relatedness of implicated genes using 250,000 PubMed abstracts. We first evaluated GRAIL by assessing its ability to identify subsets of highly related genes in common pathways from validated lipid and height SNP associations from recent genome-wide studies. We then tested GRAIL, by assessing its ability to separate true disease regions from many false positive disease regions in two separate practical applications in human genetics. First, we took 74 nominally associated Crohn's disease SNPs and applied GRAIL to identify a subset of 13 SNPs with highly related genes. Of these, ten convincingly validated in follow-up genotyping; genotyping results for the remaining three were inconclusive. Next, we applied GRAIL to 165 rare deletion events seen in schizophrenia cases (less than one third of which are contributing to disease risk). We demonstrate that GRAIL is able to identify a subset of 16 deletions containing highly related genes; many of these genes are expressed in the central nervous system and play a role in neuronal synapses. GRAIL offers a statistically robust approach to identifying functionally related genes from across multiple disease regions--that likely represent key disease pathways. An online version of this method is available for public use (http://www.broad.mit.edu/mpg/grail/). PMID- 19557191 TI - Exploring the free energy landscape: from dynamics to networks and back. AB - Knowledge of the Free Energy Landscape topology is the essential key to understanding many biochemical processes. The determination of the conformers of a protein and their basins of attraction takes a central role for studying molecular isomerization reactions. In this work, we present a novel framework to unveil the features of a Free Energy Landscape answering questions such as how many meta-stable conformers there are, what the hierarchical relationship among them is, or what the structure and kinetics of the transition paths are. Exploring the landscape by molecular dynamics simulations, the microscopic data of the trajectory are encoded into a Conformational Markov Network. The structure of this graph reveals the regions of the conformational space corresponding to the basins of attraction. In addition, handling the Conformational Markov Network, relevant kinetic magnitudes as dwell times and rate constants, or hierarchical relationships among basins, completes the global picture of the landscape. We show the power of the analysis studying a toy model of a funnel like potential and computing efficiently the conformers of a short peptide, dialanine, paving the way to a systematic study of the Free Energy Landscape in large peptides. PMID- 19557190 TI - An integrated strategy to study muscle development and myofilament structure in Caenorhabditis elegans. AB - A crucial step in the development of muscle cells in all metazoan animals is the assembly and anchorage of the sarcomere, the essential repeat unit responsible for muscle contraction. In Caenorhabditis elegans, many of the critical proteins involved in this process have been uncovered through mutational screens focusing on uncoordinated movement and embryonic arrest phenotypes. We propose that additional sarcomeric proteins exist for which there is a less severe, or entirely different, mutant phenotype produced in their absence. We have used Serial Analysis of Gene Expression (SAGE) to generate a comprehensive profile of late embryonic muscle gene expression. We generated two replicate long SAGE libraries for sorted embryonic muscle cells, identifying 7,974 protein-coding genes. A refined list of 3,577 genes expressed in muscle cells was compiled from the overlap between our SAGE data and available microarray data. Using the genes in our refined list, we have performed two separate RNA interference (RNAi) screens to identify novel genes that play a role in sarcomere assembly and/or maintenance in either embryonic or adult muscle. To identify muscle defects in embryos, we screened specifically for the Pat embryonic arrest phenotype. To visualize muscle defects in adult animals, we fed dsRNA to worms producing a GFP tagged myosin protein, thus allowing us to analyze their myofilament organization under gene knockdown conditions using fluorescence microscopy. By eliminating or severely reducing the expression of 3,300 genes using RNAi, we identified 122 genes necessary for proper myofilament organization, 108 of which are genes without a previously characterized role in muscle. Many of the genes affecting sarcomere integrity have human homologs for which little or nothing is known. PMID- 19557192 TI - Quantitative analysis of mechanisms that govern red blood cell age structure and dynamics during anaemia. AB - Mathematical modelling has proven an important tool in elucidating and quantifying mechanisms that govern the age structure and population dynamics of red blood cells (RBCs). Here we synthesise ideas from previous experimental data and the mathematical modelling literature with new data in order to test hypotheses and generate new predictions about these mechanisms. The result is a set of competing hypotheses about three intrinsic mechanisms: the feedback from circulating RBC concentration to production rate of immature RBCs (reticulocytes) in bone marrow, the release of reticulocytes from bone marrow into the circulation, and their subsequent ageing and clearance. In addition we examine two mechanisms specific to our experimental system: the effect of phenylhydrazine (PHZ) and blood sampling on RBC dynamics. We performed a set of experiments to quantify the dynamics of reticulocyte proportion, RBC concentration, and erythropoietin concentration in PHZ-induced anaemic mice. By quantifying experimental error we are able to fit and assess each hypothesis against our data and recover parameter estimates using Markov chain Monte Carlo based Bayesian inference. We find that, under normal conditions, about 3% of reticulocytes are released early from bone marrow and upon maturation all cells are released immediately. In the circulation, RBCs undergo random clearance but have a maximum lifespan of about 50 days. Under anaemic conditions reticulocyte production rate is linearly correlated with the difference between normal and anaemic RBC concentrations, and their release rate is exponentially correlated with the same. PHZ appears to age rather than kill RBCs, and younger RBCs are affected more than older RBCs. Blood sampling caused short aperiodic spikes in the proportion of reticulocytes which appear to have a different developmental pathway than normal reticulocytes. We also provide evidence of large diurnal oscillations in serum erythropoietin levels during anaemia. PMID- 19557193 TI - Optimal control predicts human performance on objects with internal degrees of freedom. AB - On a daily basis, humans interact with a vast range of objects and tools. A class of tasks, which can pose a serious challenge to our motor skills, are those that involve manipulating objects with internal degrees of freedom, such as when folding laundry or using a lasso. Here, we use the framework of optimal feedback control to make predictions of how humans should interact with such objects. We confirm the predictions experimentally in a two-dimensional object manipulation task, in which subjects learned to control six different objects with complex dynamics. We show that the non-intuitive behavior observed when controlling objects with internal degrees of freedom can be accounted for by a simple cost function representing a trade-off between effort and accuracy. In addition to using a simple linear, point-mass optimal control model, we also used an optimal control model, which considers the non-linear dynamics of the human arm. We find that the more realistic optimal control model captures aspects of the data that cannot be accounted for by the linear model or other previous theories of motor control. The results suggest that our everyday interactions with objects can be understood by optimality principles and advocate the use of more realistic optimal control models for the study of human motor neuroscience. PMID- 19557194 TI - The mysteries of chromosome evolution in gibbons: methylation is a prime suspect. PMID- 19557195 TI - Genome-wide analyses of exonic copy number variants in a family-based study point to novel autism susceptibility genes. AB - The genetics underlying the autism spectrum disorders (ASDs) is complex and remains poorly understood. Previous work has demonstrated an important role for structural variation in a subset of cases, but has lacked the resolution necessary to move beyond detection of large regions of potential interest to identification of individual genes. To pinpoint genes likely to contribute to ASD etiology, we performed high density genotyping in 912 multiplex families from the Autism Genetics Resource Exchange (AGRE) collection and contrasted results to those obtained for 1,488 healthy controls. Through prioritization of exonic deletions (eDels), exonic duplications (eDups), and whole gene duplication events (gDups), we identified more than 150 loci harboring rare variants in multiple unrelated probands, but no controls. Importantly, 27 of these were confirmed on examination of an independent replication cohort comprised of 859 cases and an additional 1,051 controls. Rare variants at known loci, including exonic deletions at NRXN1 and whole gene duplications encompassing UBE3A and several other genes in the 15q11-q13 region, were observed in the course of these analyses. Strong support was likewise observed for previously unreported genes such as BZRAP1, an adaptor molecule known to regulate synaptic transmission, with eDels or eDups observed in twelve unrelated cases but no controls (p = 2.3x10( 5)). Less is known about MDGA2, likewise observed to be case-specific (p = 1.3x10(-4)). But, it is notable that the encoded protein shows an unexpectedly high similarity to Contactin 4 (BLAST E-value = 3x10(-39)), which has also been linked to disease. That hundreds of distinct rare variants were each seen only once further highlights complexity in the ASDs and points to the continued need for larger cohorts. PMID- 19557196 TI - Evolutionary breakpoints in the gibbon suggest association between cytosine methylation and karyotype evolution. AB - Gibbon species have accumulated an unusually high number of chromosomal changes since diverging from the common hominoid ancestor 15-18 million years ago. The cause of this increased rate of chromosomal rearrangements is not known, nor is it known if genome architecture has a role. To address this question, we analyzed sequences spanning 57 breaks of synteny between northern white-cheeked gibbons (Nomascus l. leucogenys) and humans. We find that the breakpoint regions are enriched in segmental duplications and repeats, with Alu elements being the most abundant. Alus located near the gibbon breakpoints (<150 bp) have a higher CpG content than other Alus. Bisulphite allelic sequencing reveals that these gibbon Alus have a lower average density of methylated cytosine that their human orthologues. The finding of higher CpG content and lower average CpG methylation suggests that the gibbon Alu elements are epigenetically distinct from their human orthologues. The association between undermethylation and chromosomal rearrangement in gibbons suggests a correlation between epigenetic state and structural genome variation in evolution. PMID- 19557199 TI - Virus and host determinants of West Nile virus pathogenesis. PMID- 19557200 TI - Effects of dl-praeruptorin A on cultured neonatal rat ventricular cardiomyocytes with hypertrophy induced by endothelin-1. AB - The present study investigated whether dl-praeruptorin (Pd-Ia) prevents endothelin-1 (ET-1)-induced cardiomyocyte hypertrophy and the potential pathways that underlie such an effect. We assessed cardiomyocyte surface area, protein synthesis, the expression of Bax/Bcl2 and Jun genes, the expression of atrial natriuretic factor (ANF) and Ca2+/calmodulin-dependent kinase II (CaMK-II) activity in cultured neonatal rat ventricular cardiomyocytes with ET-1-induced hypertrophy. It was found that Pd-Ia decreased the surface area and protein synthesis rate in cardiomyocytes exposed to ET-1. Additionally, the expression of Bcl2 and Bax was increased in both the ET-1-exposed and Pd-Ia+ET- 1-treated groups compared with the control group, although this was not significant. In cardiomyocytes incubated with ET-1, the expression of ANF (Nppa) significantly increased relative to the control and Pd-Ia groups. The expression of Jun significantly increased in cardiomyocytes incubated with ET-1, but not in the Pd Ia group, where Jun levels were similar to those found for the control group. Moreover, it was found that Pd-Ia inhibited the ET-1-induced increase in intracellular Ca(2+) concentration. The results showed that Pd-Ia could conceivably be an effective therapeutic drug for treating the contractile defects associated with cardiac hypertrophy and failure. This activity may be associated with its Ca2+-antagonist effect and modulation of the expression of immediate early genes that play important roles in the mitogen-activated protein (MAP) kinase pathway. PMID- 19557197 TI - NRXN3 is a novel locus for waist circumference: a genome-wide association study from the CHARGE Consortium. AB - Central abdominal fat is a strong risk factor for diabetes and cardiovascular disease. To identify common variants influencing central abdominal fat, we conducted a two-stage genome-wide association analysis for waist circumference (WC). In total, three loci reached genome-wide significance. In stage 1, 31,373 individuals of Caucasian descent from eight cohort studies confirmed the role of FTO and MC4R and identified one novel locus associated with WC in the neurexin 3 gene [NRXN3 (rs10146997, p = 6.4x10(-7))]. The association with NRXN3 was confirmed in stage 2 by combining stage 1 results with those from 38,641 participants in the GIANT consortium (p = 0.009 in GIANT only, p = 5.3x10(-8) for combined analysis, n = 70,014). Mean WC increase per copy of the G allele was 0.0498 z-score units (0.65 cm). This SNP was also associated with body mass index (BMI) [p = 7.4x10(-6), 0.024 z-score units (0.10 kg/m(2)) per copy of the G allele] and the risk of obesity (odds ratio 1.13, 95% CI 1.07-1.19; p = 3.2x10( 5) per copy of the G allele). The NRXN3 gene has been previously implicated in addiction and reward behavior, lending further evidence that common forms of obesity may be a central nervous system-mediated disorder. Our findings establish that common variants in NRXN3 are associated with WC, BMI, and obesity. PMID- 19557201 TI - Skin disposition of d-camphor and l-menthol alone and together. AB - The terpenes camphor and menthol are often used in topical preparations, although some data indicate concern about their skin penetration after application in most commonly used vehicles. The cutaneous disposition of these substances applied alone and together in either an oily solution or a hydrogel was evaluated ex vivo using full human skin mounted in flow-through diffusion cells. After 0.5, 1 and 2 h of application, the skin was progressively tape-stripped into three fractions of stratum corneum (SC) and the remaining epidermis with the dermis. The content of terpenes in the skin layers was determined using gas chromatography. Different penetration into the skin layers was observed depending on the type of vehicle. The highest SC absorption was noted when terpenes were applied in hydrogel, where the total content in the SC was 200 microg/cm2 for camphor and 400 microg/cm2 for menthol, and the total skin absorption was 310 and 460 microg/cm2, respectively. The SC penetration of both terpenes from the oily solution was the same (approximately equal to 35 microg/cm2). When both terpenes were present in the hydrogel the SC absorption decreased, the amounts of camphor and menthol in the SC being 50 and 190 microg/cm2, respectively (total skin accumulation was 120 and 220 microg/cm2, respectively). Such an effect was not observed for the oily solution. PMID- 19557202 TI - Antinociceptive and anti-inflammatory effects of the ethanol extract of Alpinia conchigera Griff. leaves in various animal models. AB - The ethanolic extract of Alpinia conchigera Griff. leaves (EACL) was evaluated for its antinociceptive and anti-inflammatory activities in several in vivo experimental models. Antinociceptive activity was determined using the acetic acid-induced abdominal writhing test, the hot plate test and the formalin test. Anti-inflammatory activity was determined using the carrageenan-induced paw edema test. The extract (30, 100 and 300 mg/kg i.p.) was found to possess significant, dose-dependent inhibitory activity in all test models. In addition, the antinociceptive effect of the extract in the acetic acid-induced writhing and hot plate tests was reversed by naloxone, suggesting that this activity is mediated through activation of the opioid system. These findings suggest that EACL presents notable analgesic and anti-inflammatory activities, which support its folkloric use for painful and inflammatory conditions. PMID- 19557204 TI - Gateways to clinical trials. AB - (-)-Gossypol; Abacavir sulfate/lamivudine, ACAM-1000, ACE-011, Agomelatine, AGS 004, Alemtuzumab, Alvocidib hydrochloride, AMG-317, Amlodipine, Aripiprazole, Atazanavir sulfate, Azacitidine; Becatecarin, Belinostat, Bevacizumab, BMS 387032, BMS-690514, Bortezomib; Casopitant mesylate, Cetuximab, Choline fenofibrate, CK-1827452, Clofarabine, Conivaptan hydrochloride; Dabigatran etexilate, DADMe-Immucillin-H, Darbepoetin alfa, Darunavir, Dasatinib, DC-WT1, Decitabine, Deferasirox, Degarelix acetate, Denenicokin, Denosumab, Dienogest, Duloxetine hydrochloride; Ecogramostim, Eculizumab, Edoxaban tosilate, Elacytarabine, Elesclomol, Eltrombopag olamine, Enfuvirtide, Enzastaurin hydrochloride, Eribulin mesilate, Erlotinib hydrochloride, Escitalopram oxalate, Eszopiclone, Etravirine; Flibanserin, Fludarabine, Fondaparinux sodium, Fosamprenavir calcium; Gefitinib, Genistein; I-131-L19-SIP, Idrabiotaparinux sodium, Imatinib mesylate, IMGN-901, Ipilimumab; Laromustine, Lenalidomide, Liposomal cisplatin, Liraglutide, Lisdexamfetamine mesilate, Lopinavir, Lopinavir/ritonavir; Maraviroc, MDV-3100, Mecasermin rinfabate, MP-470, Mycophenolic acid sodium salt; Naproxcinod, NB-002, Nesiritide, Nilotinib hydrochloride monohydrate, NK-012; Palonosetron hydrochloride, Panobinostat, Pegfilgrastim, Peginterferon alfa-2a, Pitavastatin calcium, PL-3994, Plerixafor hydrochloride, Plitidepsin, PM-10450; Raltegravir potassium, Recombinant human soluble thrombomodulin, ReoT3D, RHAMM R3 peptide, Rivaroxaban, Romiplostim, Rosuvastatin calcium, Rozrolimupab; Sabarubicin hydrochloride, Salinosporamide A, Sirolimus-eluting stent, Smallpox (Vaccinia) Vaccine, Live, Sorafenib; Tenofovir disoproxil fumarate, Tenofovir disoproxil fumarate/emtricitabine, Teriparatide, Tipifarnib, Tipranavir, Trabectedin, Trifluridine/TPI; Vardenafil hydrochloride hydrate, Vinflunine, Volociximab, Vorinostat; Ximelagatran; Yttrium 90 (90Y) ibritumomab tiuxetan; Ziprasidone hydrochloride, Zoledronic acid monohydrate. PMID- 19557203 TI - Experimental rodent models of type 2 diabetes: a review. AB - Due to the high prevalence of diabetes worldwide, extensive research is still being performed to develop new antidiabetic agents and determine their mechanisms of action. Consequently, a number of diabetic animal models have been developed and improved over the years, of which rodent models are the most thoroughly described. These rodent models can be classified into two broad categories: 1) genetically induced spontaneous diabetes models; and 2) experimentally induced nonspontaneous diabetes models. The popularity of using experimentally induced nonspontaneous models for diabetes research over that of the genetically induced spontaneous models is due to their comparatively lower cost, ease of diabetes induction, ease of maintenance and wider availability. The various experimentally induced type 2 diabetes (T2D) rodent models developed over the last 30-plus years for both routine pharmacological screening and mechanistic diabetes-linked research trials include: adult streptozotocin (STZ)/alloxan rat models, neonatal STZ/alloxan models, partial pancreatectomy models, long-term high-fat (HF) diet fed models, HF diet-fed STZ models, nicotinamide/STZ models, intrauterine growth retardation (IUGR) models, the STZ-induced progressive diabetic model and monosodium glutamate (MSG)-induced model. The use of these models, however, is not without limitations. A T2D model should ideally portray an identical biochemical blood profile and pathogenesis to T2D in humans. Hence, this review will comparatively evaluate experimentally induced rodent T2D models considering the above-mentioned criteria, in order to guide diabetes research groups to more accurately select the most appropriate models given their specific research requirements. PMID- 19557205 TI - Real-life asthma care in Canada. PMID- 19557206 TI - Montelukast as add-on therapy to inhaled corticosteroids in the management of asthma (the SAS trial). AB - AIM: To evaluate the effectiveness of montelukast as add-on therapy for asthmatic patients who remain uncontrolled with low, moderate or high doses of inhaled corticosteroid monotherapy. DESIGN: An eight-week, multicentre, open-label, observational study. RESULTS: Of 320 patients enrolled, 288 (90.0%) completed the study. Of patients who had uncontrolled asthma symptoms (Canadian Asthma Consensus Guidelines Update, 2003) but were controlled according to the Asthma Control Questionnaire (ACQ score of less than 1.5), 93.9% maintained asthma control at week 8. Of patients with uncontrolled asthma at baseline for both definitions, 63.5% achieved asthma control by week 8. The mean +/- SD ACQ score decreased from 1.13+/-0.28 to 0.57+/-0.50 (P<0.001) for controlled patients at baseline and from 2.38+/-0.73 to 1.03+/-0.80 (P<0.001) for patients who were uncontrolled at baseline, each representing a clinically significant improvement. CONCLUSION: Montelukast add-on therapy is an effective alternative to inhaled corticosteroid monotherapy. PMID- 19557207 TI - Montelukast as an alternative to low-dose inhaled corticosteroids in the management of mild asthma (the SIMPLE trial): an open-label effectiveness trial. AB - OBJECTIVE: To evaluate the effectiveness of montelukast as monotherapy for patients with mild asthma who remain uncontrolled or unsatisfied while on inhaled corticosteroid (ICS) monotherapy. DESIGN: A multicentre, open-label study. Patients (six years of age or older) had ICS therapy discontinued and were treated with orally administered montelukast once daily for six weeks. MAIN OUTCOME MEASURES: The primary outcome measure was the rate at which asthma symptom control was achieved or maintained after six weeks of treatment. The secondary outcome measures were to compare compliance and physician satisfaction, and to further assess the safety and tolerability of montelukast. RESULTS: Of the 534 patients enrolled, 481 (90.1%) completed the study. Mean (+/- SD) age was 27.8+/-19.0 years. The number of patients with uncontrolled symptoms decreased from 455 (85.2%) at baseline to 143 (26.8%) at week 6 (P<0.001), and mean Asthma Control Questionnaire score decreased from 1.4+/-0.8 to 0.6+/-0.6 (P<0.001), representing a clinically significant improvement. Of the 79 patients with controlled asthma symptoms at baseline, 73.4% maintained asthma control at week 6. Compliance to asthma therapy increased from 41% at baseline for ICS to 88% at week 6 for montelukast (P<0.001). Physician satisfaction with treatment increased from 43% to 85% (P<0.001) and patient satisfaction increased from 45% at baseline to 94% at week 6. No serious adverse events were reported over the course of the study. CONCLUSION: Montelukast is an effective and well-tolerated alternative to ICS treatment in patients with mild asthma who are uncontrolled or unsatisfied with low-dose ICS therapy. PMID- 19557208 TI - Montelukast as add-on therapy with inhaled corticosteroids alone or inhaled corticosteroids and long-acting beta-2-agonists in the management of patients diagnosed with asthma and concurrent allergic rhinitis (the RADAR trial). AB - OBJECTIVE: To evaluate the effectiveness of montelukast as add-on therapy for patients diagnosed with asthma and concurrent allergic rhinitis who remain uncontrolled while receiving inhaled corticosteroid (ICS) monotherapy or ICS/long acting beta-2-agonist (LABA) therapy in a community practice setting. DESIGN: An eight-week, multicentre, open-label, observational study. Patients were 15 years of age or older and, while treated with an ICS or ICS/LABA, had allergic rhinitis and uncontrolled asthma symptoms by at least two criteria as per the Canadian Asthma Consensus Guidelines. The primary outcome measure was the percentage of patients with controlled asthma symptoms after eight weeks of treatment with montelukast 10 mg once daily added to ICS or ICS/LABA therapy. RESULTS: In total, 1004 patients participated in the survey phase of the study. Of these patients, 319 continued in the treatment phase and 301 (94.4%) completed the eight-week assessment. At baseline, all patients had uncontrolled asthma symptoms based on the Canadian Asthma Consensus Guidelines; at the eight-week assessment, 229 patients (76.1%) achieved asthma control. According to the Asthma Control Questionnaire (as determined by scores of 0.75 or less), 164 patients (54.7%) achieved well-controlled asthma at week 8. The mean (+/- SD) Asthma Control Questionnaire score decreased from 2.03+/-0.80 to 0.92+/-0.80 (P<0.001) for all patients, representing a clinically significant improvement. A statistically and clinically significant reduction in the overall Mini Rhinitis Quality of Life Questionnaire score was achieved with a decrease from 2.57+/-1.20 to 1.12+/-1.00 (-1.45+/-1.35; P<0.001). Patient and physician satisfaction rates with montelukast add-on therapy were also significantly increased when compared with baseline treatment. CONCLUSION: Montelukast add-on therapy is effective for managing asthma and allergic rhinitis symptoms in patients who were previously uncontrolled with ICS or ICS/LABA treatment. PMID- 19557209 TI - Effects of the indoor environment on the fraction of exhaled nitric oxide in school-aged children. AB - BACKGROUND: The fractional concentration of exhaled nitric oxide (FeNO) appears to be a good marker for airway inflammation in children with asthma. OBJECTIVE: To evaluate the effect of environmental exposures on exhaled nitric oxide in a community sample of children. METHODS: The relationship among exhaled nitric oxide, underlying disease and home environmental exposures was examined using questionnaire data and measurement of exhaled nitric oxide in a cross-sectional study of 1135 children that included healthy children, and children with allergies and/or asthma who were attending grades 4 through 6 in Windsor, Ontario. RESULTS: Among healthy children, there was a positive association between FeNO and occupancy (P<0.02). Compared with forced air and hot water radiant heat, electric baseboard heating was associated with a significant increase of FeNO in healthy children (P=0.007) and children with allergies (P=0.043). FeNO was not associated with environmental tobacco smoke exposure or reported surface mold. The presence of pet dog(s), but not cats, was associated with a significantly lower FeNO in healthy children (P<0.001) and in children with reported allergies (P<0.001). CONCLUSIONS: The type of heating system, but not previously reported environmental tobacco smoke or mold exposure appears to affect exhaled nitric oxide in children. Exposure to different types of pets may have disparate effects on airway inflammation. PMID- 19557212 TI - Vitamin D: the new panacea? PMID- 19557211 TI - Updating the evidence-base for suctioning adult patients: a systematic review. AB - OBJECTIVES: To update a previous clinical practice guideline on suctioning in adult patients, published in the Canadian Respiratory Journal in 2001. METHODS: A primary search of the MEDLINE (from 1998), CINAHL, EMBASE and The Cochrane Library (all from 1996) databases up to November 2007, was conducted. These dates reflect the search limits reached in the previous clinical practice guideline. A secondary search of the reference lists of retrieved articles was also performed. Two reviewers independently appraised each study before meeting to reach consensus. Study quality was evaluated using the Jadad and PEDro scales. When sufficient data were available, a meta-analysis was conducted using a random effects model. Data are reported as ORs, weighted mean differences and 95% CIs. When no comparisons were possible, qualitative analyses of the data were completed. RESULTS: Eighty-one studies were critically appraised from a pool of 123. A total of 28 randomized controlled trials or randomized crossover studies were accepted for inclusion. Meta-analysis was possible for open versus closed suctioning only. Recommendations from 2001 with respect to hyperoxygenation, hyperinflation, use of a ventilator circuit adaptor and subglottic suctioning were confirmed. New evidence was identified with respect to indications for suctioning, open suction versus closed suction systems, use of medications and infection control. CONCLUSIONS: While new evidence continues to be varied in strength, and is still lacking in some areas of suctioning practice, the evidence base has improved since 2001. Members of the health care team should incorporate this evidence into their practice. PMID- 19557213 TI - Vitamin D deficiency and chronic lung disease. AB - Vitamin D deficiency is increasingly being recognized as a prevalent problem in the general population. Patients with chronic lung diseases such as asthma, cystic fibrosis, chronic obstructive lung disease and interstitial pneumonia appear to be at increased risk for vitamin D deficiency for reasons that are not clear. Several studies indicate that vitamin D possesses a range of anti inflammatory properties and may be involved in processes other than the previously believed functions of calcium and phosphate homeostasis. Various cytokines, cellular elements, oxidative stress and protease/antiprotease levels appear to affect lung fibroproliferation, remodelling and function, which may be influenced by vitamin D levels. Chronic lung diseases such as asthma and chronic obstructive lung disease have also been linked to vitamin D on a genetic basis. This immune and genetic influence of vitamin D may influence the pathogenesis of chronic lung diseases. A recent observational study notes a significant association between vitamin D deficiency and decreased pulmonary function tests in a large ambulatory population. The present review will examine the current literature regarding vitamin D deficiency, its prevalence in patients with chronic lung disease, vitamin D anti-inflammatory properties and the role of vitamin D in pulmonary function. PMID- 19557214 TI - Workshops to disseminate the Canadian Thoracic Society guidelines for chronic obstructive pulmonary disease to health care professionals in Ontario: impact on knowledge, perceived health care practices and participant satisfaction. AB - BACKGROUND: The Canadian Thoracic Society (CTS) has developed a clinical practice guideline (CPG) regarding the management of patients with chronic obstructive pulmonary disease (COPD). Implementation of this CPG in the primary care setting requires an effective dissemination strategy. OBJECTIVES: To examine the change in knowledge, participant satisfaction and perceived changes in clinical practice among health care professionals working in the primary care setting following attendance at a workshop to disseminate the CTS CPG for COPD. METHODS: A 2.5 h workshop was conducted in three community health sites within Ontario. Each workshop comprised a didactic presentation and interactive case study discussions. Before, and one month following the workshop, a structured knowledge assessment questionnaire was administered. A structured satisfaction questionnaire and evaluative form that examined the impact of the workshop on the clinical management of COPD patients were administered immediately and three months following completion of the workshop, respectively. RESULTS: Sixty-nine participants attended the workshop. The mean score for the structured knowledge assessment questionnaire increased from 8.5+/-2.7 to 10.6+/-2.0 following the workshop (P=0.008). Eighty-nine per cent and 96% of participants indicated that they would recommend the workshop to a colleague and had greater confidence in their management of COPD patients, respectively. Following attendance of the workshop, 73%, 69% and 46% described increased patient education, patient monitoring and the use of objective testing in clinical practice, respectively. CONCLUSIONS: Workshop attendance was associated with high levels of satisfaction and important self-reported changes in clinical practice, which may reflect improved knowledge of the CTS CPG for COPD. PMID- 19557215 TI - Physical therapy practice patterns in acute exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND AND OBJECTIVE: The importance of the multidisciplinary approach to the management of chronic obstructive pulmonary disease is increasingly emphasized. The present study aimed to examine the current practice patterns of physical therapists involved in the management of patients hospitalized with an acute exacerbation of chronic obstructive pulmonary disease. METHOD: A self administered postal survey was distributed to the rehabilitation departments of all Canadian acute care hospitals with more than 250 beds. The survey addressed patient assessment, treatment, education and discharge planning for intensive care unit and/or ward admissions. RESULTS: In total, 66% of hospitals (n=109) participated in the study, with provincial representation mainly from Ontario (n=36 [33%]) and Quebec (n=36 [33%]). Assessment and treatment techniques did not differ greatly between the ward and the intensive care unit. Assessment focused on patient observation, pulse oximetry and auscultation, and was reported to be used 'always or frequently' by 76%, 76% and 69% of respondents, respectively. Less than 18% of respondents used a measure of functional capacity, and health related quality of life measures were rarely used. Treatment focused on ambulatory techniques, with 78% and 75% of respondents using mobility and transfer training 'always or frequently', respectively. The most common educational topics were breathing exercises and positioning, which were addressed by 68% and 67% of respondents, respectively. CONCLUSION: Patient assessment focused on physical impairments, with little use of measures of function or health-related quality of life, whereas treatment focused on mobility, with a lesser focus on airway clearance. Further study is needed to identify the factors that impact these practice patterns. PMID- 19557216 TI - Huge pulmonary artery aneurysm. AB - Pulmonary artery aneurysms (PAAs) are uncommon entities. PAAs are caused mostly by trauma (often iatrogenic), infections and Behcet's disease (BD). Less common causes are pulmonary hypertension, congenital heart disease and neoplasm. BD is a multisystem disorder presenting with recurrent oral and genital ulcerations, as well as ocular involvement, and PAA is one of its rare complications. A case of huge PAA, in which the usual criteria for the clinical diagnosis of BD were present, is described. Transcatheter embolization resulted in clinical improvement. PMID- 19557217 TI - The efficacy of tiotropium as a steroid-sparing agent in severe asthma. AB - People with severe asthma account for 5% to 10% of all asthmatic patients; however, this small group uses the majority of health care resources. Novel methods are needed to cope with the burden that this minority of patients places on the health care system. A severe asthma clinic patient, who was monitored through the University of Alberta's Virtual Asthma Clinic (Edmonton, Alberta) is presented. Despite optimization of his disease and individualized asthma education (provided by a certified asthma educator), the patient remained on oral glucocorticosteroids (OGS) to control his disease. Following optimization and stabilization, a further reduction in the dose of his OGS by the addition of the long-acting anticholinergic agent tiotropium bromide, was demonstrated. The role of tiotropium as a potential 'steroid-sparing agent' in severe refractory asthma is discussed, noting that if patients who are on OGS are not monitored for active inflammation, they may overuse the amount of prescribed systemic steroids, which can result in long-term steroid-related sequelae. PMID- 19557219 TI - Comparison of sediment quality guidelines (SQGs) for the assessment of metal contamination in marine and estuarine environments. AB - Sediment quality guidelines (SQGs) are an important tool for the assessment of contamination in marine and estuarine sediments. Although such guidelines are not definitive indicators of toxicity, they can have a high predictive ability and are a vital tool for identifying areas with potentially adverse biological effects. In the present study, 15 sets of common SQGs have been compared, including values for Australia/New Zealand, Canada, Hong Kong, Norway, the Netherlands, the USA and regions within the USA (Puget Sound/Washington, New York and Florida). The majority of these SQGs are based on the weight-of-evidence approach. In particular, the sub-group of TEL/PEL-based values have a very high degree of comparability; values not belonging to this uniform group show substantial variations. PMID- 19557221 TI - Quality assurance in ecological monitoring--towards a unifying perspective. PMID- 19557220 TI - Determination of carbamazepine in aquatic organisms by liquid-liquid extraction and liquid chromatography-tandem mass spectrometry. AB - A sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the determination of carbamazepine in microgram quantities of crustacean Thamnocephalus platyurus and the algae Pseudokirchneriella subcapitata at the ng L(-1) level. This fully validated method applied to a bioaccumulation study was found suitable for the detection of carbamazepine in small aquatic tissues. PMID- 19557222 TI - Rapid biodiversity assessment in lichen diversity surveys: implications for quality assurance. AB - Rapid Biodiversity Assessments (RBAs) of lichen communities, obtained by means of simplified sampling lists based on morphospecies, showed good correlations with Lichen Diversity Values (LDVs), based on the complete identification of lichen species only when performed by operators with high levels of taxonomic knowledge. Furthermore, the use of highly simplified sampling lists did not lead to significant advantages in terms of time needed for field operations. This approach proved to be especially unreliable in high diversity ecological contexts where variation of morpho-structural composition within lichen communities is frequent (i.e. co-occurring crustose- and foliose-dominated communities); it may also lead to weak results if applied for conservation purposes. Hence, the use of simplified RBA sampling lists in lichen monitoring has to be carefully evaluated and, in any case, should be based on sound taxonomic knowledge on the part of those in charge of data collection. The proper assessment of descriptors of lichen abundance and/or frequency, however, strictly depends on the skill, taxonomic knowledge, and willingness to learn of the lichenologist-in-training. PMID- 19557223 TI - Non-sampling error in ozone biomonitoring: the role of operator training. AB - Two groups assessed ozone symptoms on tobacco leaves: one was represented by young students and the other by scientists with experience in plant biology, but not experienced scorers. In the first case, results demonstrate that in the first week of exposure the extent of injury is almost always overestimated, but in the second week it is correctly evaluated or slightly underestimated: this can be due to the variable ambient ozone levels. In the second case, the average accuracy levels ranged from 40 to 82%, with an average repeatability of 95.2%. Central classes of damage are more difficult to evaluate: this may depend on the fact that two leaves may have similar total injured area, but substantially different number and spatial distribution of the lesions. Some practical suggestions in order to reduce non-sampling errors and to improve operator training are given. PMID- 19557225 TI - Sampling bias and sampling errors in pollen counting in aerobiological monitoring in Italy. AB - Information about airborne pollen concentration is of concern for health authorities across Europe. The reliability of data estimates depends on the accuracy and precision of pollen counts. In Italy, pollen counts are carried out on slides for microscopic evaluation and are regulated by the national Standard UNI 11108:2004. Our results showed that counts performed according to the Italian standard may result in a significant bias in the number of pollen grains counted and this will have an impact on final estimates of pollen concentration. For the same sample size, confidence intervals vary in relation to pollen abundance, either in terms of number of grains or of number of species. The sample size suggested by the standard (20% of the target surface) may result in errors in pollen counts ranging from 7-55% of the mean value, and in missing 22-54% of the taxa present on the slide. PMID- 19557226 TI - The running waters macroinvertebrates community: sampling techniques. AB - The community of running water macroinvertebrates has proved to be one of key subjects for fluvial ecology and bioindication studies, thanks both to the different trophic roles within the range of taxa and to the ease with which they may be collected and identified. However, the complex nature of this community creates problems concerning the complete identification of the full range of taxa, even when restricting the taxonomic classification to families and genera. Even so, the need to use the community for the implementation of indexes of Ecological Status of freshwaters and for the detection of reference conditions, necessarily means a deeper knowledge of this structure. Hence, a standard methodology of the capture effort is required to identify not only the ecological quality but also a reference community for each selected fluvial typology and for each section examined. Starting from the processing of data collected during intercalibration exercises of the IBE method, the authors analyse the results underlining the share given by the size of the sample collected (catchment effort), and by the distribution models of different taxa within the community, in order to give a contribution to the evaluation of the reliability level of standard samples. The results confirm the models already described in previous publications and lead us to accept the presence of marginal degrees of uncertainty in standard samples. PMID- 19557227 TI - Quality control procedures in the Italian national forest inventory. AB - National forest inventories represent a fundamental source of data and knowledge for forestry and environmental policy and allow for the production of national and regional level statistics on forests. The value of these statistics confirms the need for a sampling design that adequately delivers representation by reducing sampling error, but also for a data quality process that limits the non sampling errors. The article summarizes the quality control procedures of the three sampling phases adopted in the Italian national forest inventory, carried out between 2003 and 2006. The development of an integrated system of actions and controls which are able to limit subjective interpretations, in order to guarantee harmonized information all over the country, was a considerable effort within the overall project. Critical points to be considered were the consistent number of measures and evaluations undertaken during the three inventory phases, the high variability of observed attributes, the consistent number of surveyors involved, and costs of quality control, especially those related to fieldwork. At the end, examples on the overall quality of the classification performed on land cover and vegetation are discussed, as well as the impact of classification errors on the total forest area estimates. PMID- 19557224 TI - Influence of QA/QC procedures on non-sampling error in deposition monitoring in forests. AB - A Working Ring Test (WRT) was organised in the framework of the EU Regulation (EC) No 2152/2003 ("Forest Focus") and of the UN/ECE Program "ICP Forests" to evaluate the overall performance of the laboratories monitoring atmospheric deposition and soil solution in European Forests. Seven natural samples of atmospheric deposition and soil solutions and 5 synthetic solutions were distributed to 52 laboratories, which analysed them using their routine methods. Thirteen variables are considered in this paper: pH, conductivity, calcium, magnesium, sodium, potassium, ammonium, sulfate, nitrate, chloride, total alkalinity, total dissolved nitrogen and dissolved organic carbon. For each variable, the relative standard deviation of the results was evaluated, after outlier rejection, to estimate the analytical error of the measurements. The results are evaluated considering the Quality Assurance/Quality Control (QA/QC) procedure included in the ICP Forests monitoring manual: consistency check of the data and use of control charts and internal standards. A Data Quality Objective (DQO) is defined for each of the variables and the number of data meeting the DQOs are discussed in relation to the QA/QC procedures adopted. Although 38% of the results did not meet the DQO, the laboratories adopting QA/QC procedures produced a larger proportion of results meeting the objective and a consistent part of the outliers could be detected a posteriori checking analyses consistence. PMID- 19557229 TI - Applying quality assurance procedures to environmental monitoring data: a case study. AB - Managing data in the context of environmental monitoring is associated with a number of particular difficulties. These can be broadly split into issues originating from the inherent heterogeneity of the parameters sampled, problems related to the long time scale of most monitoring programmes and situations that arise when attempting to maximise cost-effectiveness. The complexity of environmental systems is reflected in the considerable effort and cost required to collect good quality data describing the influencing factors that can improve our understanding of the interrelationships and allow us to draw conclusions about how changes will affect the systems. The resulting information is also frequently elaborate, costly and irreplaceable. Since the quality of the results obtained from analysing the data can only be as good as the data, proper management practices should be considered at all stages of the monitoring activity, if the value of the information is to be properly exploited. Using a Quality Assurance system can aid considerably in improving the overall quality of a database, and good metadata will help in the interpretation of the results. The benefits of implementing Quality Assurance principles to project management and data validation are demonstrated for the information collected for the long-term monitoring of the effects of air pollution on the forest environment under Forest Focus. However, there are limits in the ability of any computer system to detect erroneous or poor quality data, and the best approach is to minimise errors at the collection phase of the project as far as possible. PMID- 19557228 TI - Quality Assurance and measurement errors in monitoring tree crown conditions in Italy. AB - The adoption of a comprehensive Quality Assurance (QA) programme for tree condition monitoring resulted in a rapid and steady improvement of data consistency in crown transparency assessment. On the other hand, the assessment of damage due to insects and fungi remains problematic and requires further training and control efforts. While our findings demonstrated the benefits arising from a QA programme and even the need to reinforce it, recent financial constrains have resulted in a severe reduction of the field checks. This will render it difficult to obtain a sound estimate of data quality and will jeopardize the results of any statistical analysis aimed at identifying status and trends of tree conditions in Italy. PMID- 19557230 TI - ICP-Forests (International Co-operative Programme on Assessment and Monitoring of Air Pollution Effects on Forests): Quality Assurance procedure in plant diversity monitoring. AB - Knowledge of accuracy and precision rates is particularly important for long-term studies. Vegetation assessments include many sources of error related to overlooking and misidentification, that are usually influenced by some factors, such as cover estimate subjectivity, observer biased species lists and experience of the botanist. The vegetation assessment protocol adopted in the Italian forest monitoring programme (CONECOFOR) contains a Quality Assurance programme. The paper presents the different phases of QA, separates the 5 main critical points of the whole protocol as sources of random or systematic errors. Examples of Measurement Quality Objectives (MQOs) expressed as Data Quality Limits (DQLs) are given for vascular plant cover estimates, in order to establish the reproducibility of the data. Quality control activities were used to determine the "distance" between the surveyor teams and the control team. Selected data were acquired during the training and inter-calibration courses. In particular, an index of average cover by species groups was used to evaluate the random error (CV 4%) as the dispersion around the "true values" of the control team. The systematic error in the evaluation of species composition, caused by overlooking or misidentification of species, was calculated following the pseudo-turnover rate; detailed species censuses on smaller sampling units were accepted as the pseudo-turnover which always fell below the 25% established threshold; species density scores recorded at community level (100 m(2) surface) rarely exceeded that limit. PMID- 19557231 TI - An experience of data quality evaluation in pollen monitoring activities. AB - In this work we report the results of some quality control tests performed on laboratory procedures referring to aerobiological monitoring activities between 2005 and 2007. In every test, the results of each operator have been compared with the scores of a group of experts. For quality evaluation, we have used accuracy and precision to define the percentage error for identification of taxa and counts of pollen grains. Cohen's K has been estimated for the analysis of reliability. This work suggests a method to set up an intercalibration test for airborne pollen monitoring and to introduce the suitableness of measurement quality objectives (MQOs). The results show the important role of operators' training and the need for standards in pollen monitoring quality evaluation. PMID- 19557232 TI - Validation of air pollution biomonitoring networks and related data modelling: a geostatistical approach. AB - This paper describes a geostatistical approach for environmental data modeling based on value descriptive analysis, variographic analysis, estimation method and validation. It shows an application related to atmospheric pollutant dispersion (ozone) detected by active biomonitoring in the province of Livorno (Italy). This work proposes a two-stage approach based on: the computation of an index on an exploratory and geostatistical data analysis to assess whether a monitoring network is well distributed in space and allows us to get reliable information about the whole study area; the derivation of a continuous representation of the variable (the Cotyledonous Damage Index) from punctual measurements. The geostatistical approach proposed is useful to define in a structured way the possible problems in a monitoring network, to control the data estimation error in the points not sampled. The case study analysed underlines that the biomonitoring network outline does not have a representative sample station distribution of the study area introducing significant errors related to the territorial generalization of the derived information. The proposed approach allows us to elaborate on the obtained data to carry out the data spatialization associating an error to them and to the evaluation model. PMID- 19557233 TI - Using taxonomic data to assess and monitor biodiversity: are the tribes still fighting? AB - In this article we review the problems encountered during the use of taxonomic information for the purpose of monitoring biodiversity. These problems encompass the nature of taxonomic data that requires human interpretation in order to be recognised in the field and grouped into well-defined classes such as species. We then briefly discuss some methods that may be utilized in order to minimise these problems. PMID- 19557234 TI - Assessing inter-generational transfer of a brominated flame retardant. AB - Studies have shown that the lipophilic nature of polybrominated biphenyl (PBB) causes it to preferentially accumulate in breast milk posing a potential hazard for suckling infants. The purpose of this study was to examine the inter generational transfer of PBB from mother to child and whether this association was modified by maternal breast-feeding patterns. One hundred and forty-five mother-child pairs that were participants of the Michigan Long-Term PBB Study were included in this analysis. Mothers were exposed to PBB via contaminated food between 1973 and 1974 and children were exposed in utero and for some, through breast-feeding. Seventy-three percent of children had a non-detectable serum PBB concentration (limit of detection (LOD) = 1 microg L(-1)). Mothers' serum PBB concentration at enrollment ranged from or =8 microg L(-1), breast-feeding > or =5.5 months, maternal age at child's birth > or =28 years, and being born during the PBB exposure period. Among mothers with a detectable serum PBB concentration, those who breast-fed > or =5.5 months were 6 times more likely to have a child with a detectable serum PBB concentration, compared to a non-breast-fed child (95% C.I., 2.0-19.6). PMID- 19557235 TI - Fate and transport of Irgarol 1051 in a modular estuarine mesocosm. AB - Fate and transport of Irgarol 1051 were assessed using a modular estuarine mesocosm containing natural seawater, saltmarsh sediments, marsh grass, shrimp, clams, snails, and naturally derived planktonic and benthic microorganisms. The mesocosms were enclosed in a greenhouse under near ambient conditions, and included a saltwater sump and a simulated tidal flux. The exposure was comprised of four replicate treatments of 0, 100, 1,000 and 10,000 ng/L Irgarol. Solid phase extraction (SPE) was used to extract Irgarol and its major metabolite M1 (aka GS26575) from water samples. Sediment samples were extracted using Accelerated Solvent Extraction (ASE) with 100% dichloromethane. Irgarol and M1 were separated and quantified utilizing Liquid Chromatography Tandem Mass Spectrometry (LC-MS-MS) with ElectroSpray Ionization in Multiple Reaction Monitoring mode. Aqueous concentrations of Irgarol declined rapidly (average 93% loss) over the course of the 35 day experiment while Irgarol was accumulated in the sediments (average mass balance of 75 +/- 5%). Loss of aqueous Irgarol occurred in two distinct phases, a relatively rapid phase up to 96 hours post dose, and a slower phase following 96 hours. The rate constants of the initial rapid degradation phase (k(1)) for treatments 100 and 1,000 ng/L Irgarol were 2-3 times higher than the rate constants of the subsequent slow degradation phase (k(2)) for these treatments. The average half-life of Irgarol in water was 7 +/- 3 days. The aqueous concentration of Irgarol's metabolite, M1 increased over the course of the experiment. By 35 days post-dose, M1 concentrations in water averaged about 3% of the parent compound. M1 also accumulated in mesocosm sediments (average 16.7 +/- 2.5% of total Irgarol dose). Mass balance calculations showed that by 35 days, Irgarol and M1 amounts in mesocosm water and sediments were close to 100% of the initial Irgarol dose. Average partition coefficient normalized for organic carbon (log K(oc)) calculated for Irgarol was 3.2 +/- 0.1 for 35 days post-dose. Accumulation of Irgarol and M1 in mesocosm sediments may warrant further study to assess toxicity for benthic communities. PMID- 19557236 TI - Models for the sorption of volatile organic compounds by diesel soot and atmospheric aerosols. AB - The solvation parameter model is used to characterize interactions responsible for the sorption of varied organic compounds by diesel soot and atmospheric aerosols at 15 degrees C and 50% relative humidity. Individual models are obtained for eight aerosol samples characterized as urban, suburban, rural and coastal. Combining the individual aerosol models resulted in a general aerosol model with only a minor loss of modeling power for alkanecarboxylic acids and low molecular weight alcohols compared with the individual models. A second group of compounds identified as weak nitrogen-containing bases were consistent outliers to all models most likely due to participation in ion-exchange interactions not considered by the models. The diesel soot and atmospheric aerosols exhibit similar characteristics with respect to their sorption interactions although differences in relative magnitude allow the two particle types to be easily distinguished. Sorption interactions are favored by strong dispersion interactions for both particle types. Of note is the strong hydrogen-bond basicity and relatively weak hydrogen-bond acidity of these materials. The particles are quite dipolar/polarizable and slightly electron lone pair repulsive. The sorption properties of the atmospheric aerosols are influenced by the relative humidity, in particular, the aerosols become significantly more hydrogen-bond acidic at high relative humidity most likely due to incorporation of increasing amounts of condensed or film water in the aerosol phase. Dividing the data into training and test sets suggests that the proposed models are capable of estimating distribution constants (log K) to about 0.20 log units for diesel soot (n = 84) and 0.14 log units for the general atmospheric aerosol model (n = 385) where n indicates the number of compounds included in the model. PMID- 19557237 TI - Comparison of nickel release in solutions used for the identification of water soluble nickel exposures and in synthetic lung fluids. AB - Chemical speciation of workplace nickel exposures is critical because nickel containing substances often differ in toxicological properties. Exposure matrices based on leaching methods have been used to ascertain which chemical forms of nickel are primarily associated with adverse respiratory effects after inhalation. Misjudgments in the relative proportion of each of the main fractions of nickel in workplace exposures could translate into possible misattributions of risk to the various forms of nickel. This preliminary study looked at the efficiency of the first step of the Zatka leaching method for accurately assessing the 'water-soluble' fraction of several substances present in nickel production operations, compared to leaching in synthetic lung fluid. The present results demonstrate that for nickel sulfate or chloride, the current Zatka solution is adequate to assess the 'water-soluble' fraction. However, when sparingly water-soluble compounds like nickel carbonates or water-insoluble substances like nickel subsulfide and fine metallic nickel powders are present, the first step of the Zatka method can greatly over estimate the amount of nickel that could be released in pure water. In contrast, the releases of nickel from nickel carbonate, nickel subsulfide, and nickel metal powders in pure water are consistent with their releases in synthetic lung fluid, indicating that deionized water is a better leaching solution to estimate the biologically relevant 'water soluble' nickel fraction of workplace exposures. Exposure matrices relying mostly on the Zatka speciation method to estimate the main forms of nickel need to be re evaluated to account for any possible misattributions of risk. PMID- 19557238 TI - Determination of bezafibrate, methotrexate, cyclophosphamide, orlistat and enalapril in waste and surface waters using on-line solid-phase extraction liquid chromatography coupled to polarity-switching electrospray tandem mass spectrometry. AB - We developed a rapid method for the monitoring of five selected pharmaceuticals in the influent and effluent of municipal wastewater treatment plants (WWTP) as well as in the effluent-receiving waters. To that end, we optimized and validated an analytical method based on on-line solid-phase extraction (on-line SPE) coupled with reversed-phase liquid chromatography-switching polarity electrospray ionization-tandem mass spectrometry (LC-ESI(+/-)-MS/MS). The target analytes have a variable hydrophobic character and belong to various therapeutic classes including the lipid regulator bezafibrate, the chemotherapy drugs methotrexate and cyclophosphamide, the lipase inhibitor orlistat and the angiotensin converting enzyme (ACE) inhibitor used in the treatment of hypertension, enalapril. The method combines positive and negative voltage switching modes, therefore all analytes can be determined using a single injection and without any reduction in sensitivity. In order to detect traces of these compounds, a preconcentration step before detection is performed by loading 1.00 mL of sample in an on-line SPE cartridge and eluting from the cartridge using a reversed-phase liquid chromatography gradient. Analysis of wastewater and surface water samples was greatly affected by co-eluting matrix compounds, to compensate for matrix effects quantitation was therefore performed using standard additions. Method intra-day precision was less than 6.5% and limits of detection in fortified matrix effluent samples ranged from 9 to 20 ng L(-1). Four of the target pharmaceuticals were detected in the WWTP effluents, enalapril and bezafibrate being the most abundant compounds with concentrations of 35 and 239 ng L(-1), respectively. Concentrations of these same compounds in surface water samples from sites downstream in the St. Lawrence River were 8 and 63 ng L(-1), respectively, which was mainly due to dilution. PMID- 19557239 TI - Precipitation chemistry in Croatia during the period 1981-2006. AB - The spatial, temporal changes and trends changes in precipitation chemistry in Croatia during the period 1981-2006 were analysed using data from 27 sampling sites. The origins of air masses that arrived over Croatia were evaluated by the sector analysis of two-dimensional back trajectories. The distribution per sector of the volume-weighted concentrations of the main ions (sulfate, nitrate, chloride, ammonium, calcium, magnesium, sodium, and potassium) and the pH were also inspected. The trend in the annual volume-weighted concentrations of the major ions in precipitation and pH was tested using the nonparametric Mann Kendall test and Sen's slope estimator. The sulfate and nitrate ion concentrations significantly decreased for the majority of sites. The main decline of the sulfate ion concentration occurred after 1990. Changes in the anthropogenic sulfur emission patterns over Europe in the last two decades resulted in different acidity and sulfate ion concentrations related to a particular sector compared to earlier times. In addition, precipitation chemistry in Croatia was also influenced by geographical and climatological diversity. Finally, the war and post-war activities were identified as possible anthropogenic sources of some base cations, i.e., potassium and calcium. PMID- 19557240 TI - Optical fiber analyzer for in situ determination of nitrous oxide in workplace environments. AB - An analytical method based on optical fiber (OF) detection has been developed for monitoring nitrous oxide (N(2)O) in workplace environments. The analyzer was firstly tested for calibration and then its analytical performance for N(2)O detection compared with solid phase microextraction-gas chromatography-mass spectrometry, an already validated method for determination of N(2)O. The response of the OF analyzer was linear from 0.20 to 1.80 ppb, and the detection limit was less than 0.03 ppb. After some laboratory tests, the developed analytical system was applied to air monitoring at a confined industrial environment showing a suitable compact design and a high analytical performance for N(2)O monitoring, both in situ and in a real-time mode. PMID- 19557241 TI - Design and performance evaluation of a medium flow sampler for airborne brominated flame retardants (BFRs). AB - Brominated flame retardants (BFRs), including polybrominated diphenyl ethers (PBDEs), have emerged as important and ubiquitous environmental pollutants, and there is a need to accurately measure airborne levels of these chemicals in both indoor and outdoor applications. We review and suggest performance criteria for BFR sampling systems, and then present the design of a new medium flow active sampler. The sampler uses a PTFE filter (47 mm, 1 microm pore size) in front of a polyurethane foam (PUF) adsorbent plug (22 mm dia, 76 mm length) with a nominal flow rate of 15 L min(-1) and a sampling period of one week, giving a sampling volume of 150 m(3). The sampler was evaluated using co-located systems to test precision, backup PUFs to test breakthrough, and distributed volume sampling to test linearity. Field experiments were conducted in five commercial buildings, one residence and outdoors at an urban site. A total of 20 BDE congeners were quantified. After appropriate cleaning of the PUF adsorbent, blank levels were negligible. Method detection limits (MDLs) were sufficiently low to quantify BDE congeners 17, 28, 71, 47, 100 and 99 in ambient air, and more than adequate to quantify these and other congeners in indoor air, where levels are typically much higher. The relative absolute deviation (RAD), based on distributed volume samples, ranged from 21% (BDE-71) to 81% (BDE-75) for indoor samples, and was somewhat higher for ambient samples. Only minimal breakthrough was detected in back-up samples, and over 80% of the samples had very low or negligible breakthrough. Humidity did not influence sampler performance. Overall, the medium flow sampler can accurately measure PBDEs over a wide range of concentrations and applications. PMID- 19557242 TI - Estimation and characterization of PCDD/Fs and dioxin-like PCB emission from secondary zinc and lead metallurgies in China. AB - Secondary zinc and lead production is addressed as one of the potential sources of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dioxin-like PCBs) due to the impurity of the raw material. However, the release inventory of PCDD/Fs and dioxin-like PCBs is very scarce for these secondary nonferrous metallurgies. This study selected typical secondary zinc and lead plants to investigate the emissions of PCDD/Fs and dioxin like PCBs released from such plants in China. The toxic equivalent quantity (TEQ) emission factor of PCDD/Fs and dioxin-like PCBs released into the environment is higher for secondary zinc production, at 52 298.02 ng TEQ ton(-1) than for secondary lead production, at 646.05 ng TEQ ton(-1). The most abundant congeners of PCDD/Fs are 2,3,4,7,8-PeCDF and 1,2,3,4,6,7,8-HpCDF for the secondary zinc and lead metallurgies, respectively. The most abundant congener of dioxin-like PCBs in the samples collected from both metallurgies is CB-126 . According to the distribution of PCDD/Fs (PCDF/PCDD >> 1) and the dominant contribution of higher chlorinated congeners, the de novo synthesis is assumed to be the main formation pathway of PCDD/Fs in the secondary zinc metallurgies. For the secondary lead metallurgies, the role of precursor formation is also very important. Based on the emission factor and production level, the total estimated emission amounts of PCDD/Fs and dioxin-like PCBs in both stack gas and fly ash released into the environment from secondary zinc and lead production is estimated to be at least 2.76 and 0.42 g TEQ yr(-1), respectively. The dioxin-like PCBs contribute 2.8% and 0.6% of the total emission from secondary zinc and lead plants, respectively. PMID- 19557243 TI - Presence and mobility of arsenic in estuarine wetland soils of the Scheldt estuary (Belgium). AB - We aimed to assess the presence and availability of arsenic (As) in intertidal marshes of the Scheldt estuary. Arsenic content was determined in soils sampled at 4 sampling depths in 11 marshes, together with other physicochemical characteristics. Subsequently, a greenhouse experiment was set up in which pore water arsenic (As) concentrations were measured 4 times in a 298-day period in 4 marsh soils at different sampling depths (10, 30, 60 and 90 cm) upon adjusting the water table level to 0, 40 and 80 cm below the surface of these soils. The As content in the soil varied significantly with sampling depth and location. Clay and organic matter seem to promote As accumulation in the upper soil layer (0-20 cm below the surface), whereas sulfide precipitation plays a significant role at higher sampling depths (20-100 cm below the surface). The As concentrations in the pore water of the greenhouse experiment often significantly exceeded the Flemish soil sanitation thresholds for groundwater. There were indications that As release is not only affected by the reductive dissolution of Fe/Mn oxides, but also by e.g. a direct reduction of As(V) to As(III). Below the water table, sulfide precipitation seems to lower As mobility when reducing conditions have been sufficiently established. Above the water table, sulfates and bicarbonates induce As release from the solid soil phase to the pore water. PMID- 19557244 TI - Determination of particulate and gaseous phthalic acid esters in ambient air by large-volume injection-gas chromatography-mass spectrometry. AB - A sensitive and effective analytical method has been optimized and validated for the determination of particulate and gaseous phthalic acid esters (PAEs) in ambient air. The samples were Soxhlet extracted with an n-hexane-dichloromethane methanol mixture (1:1:1, volume ratio), and analyzed using large-volume injection coupled to a gas chromatography-mass spectrometry (LVI-GC-MS) system. Standard sample linearity in the range of 0.02 approximately 5 microg L(-1) provides regression coefficients (R(2)) better than 0.995. Limits of detection (LODs) and limits of quantification (LOQs) were at the microg L(-1) level. Because the air sample volumes were 360 m(3), the accuracy and precision for the PAE concentrations at the pg m(-3) level were guaranteed. This sensitivity is much higher than the conventional 2 microL splitless injection and is suitable for the detection of ultra-trace PAEs in complex air matrices. PMID- 19557245 TI - Characterization of trace metals of risk to human health in airborne particulate matter (PM2.5) at two sites in Guadalajara, Mexico. AB - PM2.5 samples were collected at two locations in Guadalajara: Centro and Miravalle, during 2007. The first site (Centro) is located downtown and characterized by high vehicular traffic. Miravalle is in the southern part of the city, and influenced by emissions from high industrial and vehicular activity. Samples were collected for 24 h and the annual median concentrations of PM2.5 observed were 44.1 and 52.8 microg m(-3) at Centro and Miravalle, respectively. The concentration of PM2.5 observed at the Miravalle site was significantly higher (p < 0.002). Ca, Fe, Zn, Mg and Pb were the most abundant elements found at both sites. Miravalle showed higher annual concentrations of Ca, Pb, Cu, Cr, Sr, Ni, Mo, Fe, Mn, and Sb with levels of 1013.9, 74.9, 28.2, 9.4, 6.3, 4.4, 1.0, 628.0, 23.7 and 4.6 ng m(-3), respectively. At Centro and Miravalle quarterly and annual concentrations of Pb did not exceed 290 ng m(-3) and all values are well below those specified by air quality standards. The Principal Component Analysis (PCA), Cluster Analysis and the enrichment factor (EF) based on the concentrations of each element indicated that the main source of particulates at Centro was of geological origin, while Miravalle receives emissions from natural and anthropogenic sources. Both contribute to the chemical composition of PM2.5 in Guadalajara. PMID- 19557249 TI - Bicyclic-guanidines, -guanidinates and -guanidinium salts: wide ranging applications from a simple family of molecules. AB - Bicyclic guanidines have physical, electronic and chemical properties that differentiate them from their acyclic counterparts, with many of these characteristics directly imposed by the rigid framework. These distinctive features have led to this class of molecule finding practical applications in many areas of chemistry, including organocatalysis and as ligands in coordination compounds. In addition to the neutral molecules, the corresponding cationic (guanidinium) and anionic (guanidinate) species have also been widely studied. Applications of these ions range from anion recognition and supramolecular arrays involving guanidinium salts, to the utilization of bicyclic guanidinate anions as ligands for metal compounds and clusters. This article reviews the chemistry of these compounds in light of recent advances in the synthesis of new derivatives, highlighting the potential for cross-stimulation of different areas. PMID- 19557250 TI - Recent advances in the understanding of the interaction of antidepressant drugs with serotonin and norepinephrine transporters. AB - The biogenic monoamine transporters are integral membrane proteins that perform active transport of extracellular dopamine, serotonin and norepinephrine into cells. These transporters are targets for therapeutic agents such as antidepressants, as well as addictive substances such as cocaine and amphetamine. Seminal advances in the understanding of the structure and function of this transporter family have recently been accomplished by structural studies of a bacterial transporter, as well as medicinal chemistry and pharmacological studies of mammalian transporters. This feature article focuses on antidepressant drugs that act on the serotonin and/or the norepinephrine transporters. Specifically, we focus on structure-activity relationships of these drugs with emphasis on relationships between their molecular properties and the current knowledge of transporter structure. PMID- 19557252 TI - Copper(I) scorpionate complexes and their application in palladium-mediated [(11)C]carbonylation reactions. AB - Solutions of copper(I) tris(pyrazolyl)borate complexes have been used to greatly improve the solubility of [(11)C]carbon monoxide, enabling it to be used in low pressure, 'one-pot' palladium-mediated carbonylation reactions to form (11)C radiolabelled amides and ureas for use in positron emission tomography. PMID- 19557251 TI - Antibacterial cyclic D,L-alpha-glycopeptides. AB - We report the design, synthesis, membrane activity, biophysical characterization, and in vitro antibacterial activities of cationic cyclic D,L-alpha-glycopeptides bearing d-glucosamine (GlcNH(2)), D-galactose (Gal), or D-mannose (Man) glycosyl side chains. PMID- 19557253 TI - Amidinate-ligated iridium(III) bis(2-pyridyl)phenyl complex as an excellent phosphorescent material for electroluminescence devices. AB - Highly efficient, low driving-voltage, and emitter concentration insensitive phosphorescent EL devices are established for the first time by using an amidinate-ligated iridium(III) complex as an emitting component. PMID- 19557254 TI - Naphthalene-bis-hydrazimide: radical anions and ICT as new bimodal probes for differential sensing of a library of amines. AB - A new strategy of a bimodal sensing for amines and the first applications of radical anions as probes to colorimetrically differentiate a library of amines is reported. PMID- 19557255 TI - Assembling p-type molecules on single wall carbon nanotubes for photovoltaic devices. AB - We report the design and synthesis of an oligothiophene molecule that noncovalently functionalizes carbon nanotubes to create a hybrid material for photovoltaic devices. PMID- 19557256 TI - Two-phase dynamic combinatorial discovery of a spermine transporter. AB - The discovery, in a two-phase dynamic combinatorial library, of an unexpected linear receptor and transporter for spermine is described. PMID- 19557257 TI - Transport of calcium ions through a bulk membrane by use of a dynamic combinatorial library. AB - In a bulk membrane transport experiment, a dynamic combinatorial library (DCL) has been used to transport calcium ions; the calcium ions amplify the formation of a macrocyclic carrier which results in transport. PMID- 19557258 TI - Enabling olefin metathesis on proteins: chemical methods for installation of S allyl cysteine. AB - Multiple, complementary methods are reported for the chemical conversion of cysteine to S-allyl cysteine on protein surfaces, a useful transformation for the exploration of olefin metathesis on proteins. PMID- 19557259 TI - Aluminium hydride: a reversible material for hydrogen storage. AB - Aluminium hydride has been synthesized electrochemically, providing a synthetic route which closes a reversible cycle for regeneration of the material and bypasses expensive thermodynamic costs which have precluded AlH(3) from being considered as a H(2) storage material. PMID- 19557260 TI - A Zn-based, pillared paddlewheel MOF containing free carboxylic acids via covalent post-synthesis elaboration. AB - A Zn-based, mixed-ligand (pillared paddlewheel), metal-organic framework (MOF) has been covalently and quantitatively decorated with free carboxylic acids to demonstrate the utility of covalent post-synthesis modification in the construction of otherwise inaccessible carboxy-functionalized MOFs. PMID- 19557261 TI - Sacrificial layer electrophoretic deposition of free-standing multilayered nanoparticle films. AB - Sacrificial layer electrophoretic deposition (SLED) is a technique to assemble nanoparticles that yields free-standing, multilayered nanoparticle films with macroscopic lateral dimensions after the sacrificial layer is dissolved. PMID- 19557262 TI - Photosensitizer efficiency in genetically modified protein cage architectures. AB - A Ru(bpy)(3)(2+) photosensitizer was covalently linked at specific sites on the interior or exterior surface of genetic constructs of a small heat shock protein cage nanoplatform and the light activated production of singlet oxygen was characterized. PMID- 19557264 TI - The oxidation of Ni(II) N-confused porphyrins (NCPs) with azo radical initiators and an unexpected intramolecular nucleophilic substitution reaction via a proposed Ni(III) NCP intermediate. AB - The oxidation of Ni(II) N-confused porphyrins (NCPs) with azo radical initiators resulted in an unexpected intramolecular nucleophilic substitution reaction via a proposed Ni(III) NCP intermediate, which could be detected by HRMS. PMID- 19557265 TI - Photoinduced hardness change on TiO(2) single crystal surfaces. AB - The dynamic hardness of rutile TiO(2) single crystal surfaces was investigated using a nanoindentation technique and the hardness increase was observed only in the near-surface region, possibly due to the volume expansion accompanied by the photoinduced high hydrophilicity. PMID- 19557263 TI - Azide ion recognition in water-CHCl(3) using a chelating phosphonium borane as a receptor. AB - In H(2)O-CHCl(3), the phosphonium borane [ortho-(Mes(2)B)C(6)H(4)(PMePh(2))](+) selectively complexes azide anions to afford ortho (Mes(2)(N(3))B)C(6)H(4)(PMePh(2)) in which the boron-bound azide anion is stabilized by an interaction with the adjacent phosphorus atom. PMID- 19557266 TI - Bio-inspired polyene cyclization: aziridinyl polyene cyclization catalyzed by InBr(3). AB - This communication describes a highly efficient aziridinyl polyolefin cyclization catalyzed by InBr(3) to synthesize chiral terpenoid bearing a 3-amino group in the A ring; both good yields and excellent asymmetric induction were achieved. PMID- 19557267 TI - Cinnamic aldehyde derived probes for the active site labelling of pathogenesis associated enzymes. AB - Michael acceptor based natural product derived probes are selective and sensitive chemical tools for the identification and characterization of pathologically relevant enzymes in MRSA. PMID- 19557268 TI - Reduction of carbon dioxide in 1-butyl-3-methylimidazolium acetate. AB - Gaseous CO(2) is almost irreversibly absorbed by the room temperature ionic liquid 1-butyl-3-methylimidazolium acetate ([C(4)mim][Ac]) in which it undergoes a chemically irreversible, one electron electrochemical reduction, suggesting a means for the sequestration of the greenhouse gas. PMID- 19557270 TI - Main-chain NHC-palladium polymer as a recyclable self-supported catalyst in the Suzuki-Miyaura coupling of aryl chlorides in water. AB - We have demonstrated an efficient and reusable NHC-palladium polymer self supported catalyst for the Suzuki-Miyaura coupling reaction, which even works for deactivated aryl chlorides and aryl fluorides in water. PMID- 19557269 TI - Soluble iron nanoparticles as cheap and environmentally benign alkene and alkyne hydrogenation catalysts. AB - Iron nanoparticles prepared by reducing FeCl(3) with three equivalents of EtMgCl in THF are effective catalysts for the hydrogenation of alkenes and alkynes under moderate conditions. PMID- 19557271 TI - C(2)-symmetric chiral tetrathiafulvalene-bis(oxazolines) (TTF-BOX): new precursors for organic materials and electroactive metal complexes. AB - Chiral C(2)-symmetric electroactive bis(oxazolines) containing an EDT-TTF (ethylenedithio-tetrathiafulvalene) backbone, as promising multifunctional ligands and precursors for chiral molecular materials, were synthesized and structurally characterized, together with octahedral molybdenum and tungsten carbonyl complexes. PMID- 19557272 TI - Nucleic acid-induced self-assembly of a platinum(II) terpyridyl complex: detection of G-quadruplex formation and nuclease activity. AB - A simple "proof-of-principle" concept for the monitoring of G-quadruplex formation and nuclease activity based on the unique self-assembly properties of a platinum(II) terpyridyl complex, has been demonstrated. PMID- 19557274 TI - Bis-rhodium hexaphyrins: metalation of [28]hexaphyrin and a smooth Huckel aromatic-antiaromatic interconversion. AB - Metalation of [28]hexaphyrin(1.1.1.1.1.1) with [RhCl(CO)(2)](2) resulted in the formation of an antiaromatic bis-Rh(i) complex, which can be oxidized to an aromatic bis-Rh(i) complex with retention of its rectangular structure. PMID- 19557273 TI - Controlled chemical etching for silicon nanocrystals with wavelength-tunable photoluminescence. AB - The size of silicon nanocrystals is finely tuned by applying a chemical etching process, leading to differently coloured visible luminescence. PMID- 19557275 TI - Two-dimensional OPV4 self-assembly and its coadsorption with alkyl bromide: from helix to lamellar. AB - Two-dimensional self-assembly of OPV4 molecule and its coadsorption with C(18)H(37)Br are investigated by scanning tunneling microscopy (STM) on highly oriented pyrolytic graphite (HOPG) revealing a structural difference and transition from helix to lamellar. PMID- 19557276 TI - Heteroepitaxial growth of MFI zeolite over titanosilicate molecular sieves. AB - For the first time heteroepitaxial growth of MFI zeolite over ETS-4 and ETS-10 is described, and a structural model of the interface between the molecular sieves is proposed. PMID- 19557277 TI - Cycloabiesesquine A, a unique sesquiterpenoid from Abies delavayi. AB - Cycloabiesesquine A, representing a unique carbon skeleton among the sesquiterpenoids, was isolated from Abies delavayi and by extensive spectroscopic and computational methods, its structure was elucidated as (7S,8R,12R)-4,8 dihydroxy-1,3,5,10-tetraen-8,12-cyclobisabolan-9-one. PMID- 19557279 TI - Binuclear gadolinium(III) coordination complex based on bridging tetrathiafulvalenecarboxylate radical cations. AB - X-Ray structure, photophysical and magnetic properties have been explored for the binuclear gadolinium(III) coordination complex based on bridging tetrathiafulvalenecarboxylate radical cations [Gd(hfac)(3)(micro-TTF (+)COO( ))](2). PMID- 19557278 TI - Photoinduced electron transfers with carbon dots. AB - The photoluminescence in carbon dots (surface-passivated small carbon nanoparticles) could be quenched efficiently by electron acceptor or donor molecules in solution, namely that photoexcited carbon dots are both excellent electron donors and excellent electron acceptors, thus offering new opportunities for their potential uses in light energy conversion and related applications. PMID- 19557280 TI - Synthesis of hybrid Fe(3)O(4)-silica-NiO superstructures and their application as magnetically separable high-performance biocatalysts. AB - The assembly of NiO nanoparticles at the surface of magnetic silica resulted in a hybrid superstructure which has utility as a carrier for a magnetically separable biocatalyst system. PMID- 19557281 TI - Improving Grubbs' II type ruthenium catalysts by appropriately modifying the N heterocyclic carbene ligand. AB - The introduction of N-heterocyclic carbene ligands that incorporate correctly substituted naphthyl side chains leads to increased activity and stability in second generation ruthenium metathesis catalysts. PMID- 19557282 TI - High-temperature anisotropic silicon-etching steered synthesis of horizontally aligned silicon-based Zn(2)SiO(4) nanowires. AB - A high-temperature anisotropic silicon-etching strategy is demonstrated to steer the growth of the horizontally localized parallel Zn(2)SiO(4) nanowires. PMID- 19557283 TI - In situ encapsulation of Au nanoparticles in mesoporous core-shell TiO(2) microspheres with enhanced activity and durability. AB - An active and durable Au/TiO(2) photocatalyst was prepared by in situ encapsulation of Au particles into core-shell TiO(2) spheres based on consecutive solvothermal and hydrothermal treatments. PMID- 19557284 TI - A photoresponsive liquid crystal based on (1-cyclohexenyl)phenyldiazene as a close analogue of azobenzene. AB - A new photoresponsive calamitic liquid crystal molecule possessing (1 cyclohexenyl)phenyldiazene as a close analogue of azobenzene was synthesized and a comparative study with the corresponding azobenzene analogue carried out. PMID- 19557285 TI - Neutral metallacyclic rotors. AB - A prototype of a neutral Re-based metallacyclic rotor, assembled in a one-step process, possesses a para-phenylene rotor that rotates in solution with respect to the stator. PMID- 19557286 TI - Gold nanoring arrays from responsive block copolymer templates. AB - We report a pH-mediated synthetic route for the production of ordered and size tuneable arrays of gold nanorings using responsive block copolymer micelles as templates. PMID- 19557287 TI - Luminescence vapochromic properties of a platinum(II) complex with 5,5' bis(trimethylsilylethynyl)-2,2'-bipyridine. AB - A platinum(II) complex of 5,5'-bis(trimethylsilylethynyl)-2,2'-bipyridine displays unusually reversible colour and luminescence changes when exposed to specific organic vapours, correlated well with Pt...Pt distances as well as pi-pi and C-H...(C[triple bond]C) interactions in the stacking of square-planar platinum(II) moieties. PMID- 19557289 TI - Vesicles fabricated by hybrid nanoparticles. AB - Polystyrene (PS) with pendant gold nanoparticles and PS-coated Fe(3)O(4) nanoparticles self-assemble into vesicle structures with PS and PS-coated Fe(3)O(4) nanoparticles in the wall and gold nanoparticles in the shell. PMID- 19557288 TI - Two spin-competing manganese(II) coordination polymers exhibiting unusual multi step magnetization jumps. AB - Two new three-dimensional manganese(II) coordination polymers were assembled by 1,2,3-triazole-4,5-dicarboxylate, showing unusual multi-step magnetization jumps at 1.8 K due to the competing interactions between the antiferromagnetic couplings with different amplitudes. PMID- 19557292 TI - The Individualized Target HbA1c: A New Method for Improving Macrovascular Risk and Glycemia Without Hypoglycemia and Weight Gain. AB - Both the DCCT and UKPDS trials demonstrated that improved glycemic control reduces microvascular complications. Inconclusive evidence, however, has remained on the question of the effect of glycemic control on macrovascular disease (with special emphasis on cardiovascular morbidity and mortality). In the last year, the data from four large trials were published, directly addressing this question (ACCORD, ADVANCE, VADT and UKPDS-80), yet the results were conflicting. Close inspection of the structure of three of these trials (ACCORD, ADVANCE and VADT) revealed inadequacies that may explain the unfavorable results, such as the inclusion of mainly elderly patients with previous macrovascular complications. It is not surprising that intensive glycemic control resulted in a rise of hypoglycemic events yet did not decrease macrovascular morbidity or mortality in these cohorts. On the other hand, the UKPDS-80 trial, a follow-up of the original UKPDS, showed that intensive glycemic control was beneficial when initiated in newly diagnosed patients. These results led us to develop a new individualized method of determining the target HbA1c based on the characteristics of the individual. This method considers the patient's possible benefit from glycemic control, the risk of suffering hypoglycemic events and consequences suffered from the hypoglycemic event. It is essential that the target HbA1c be tailored to the patient, with different goals set for the recently diagnosed "healthy" and young patient on the one hand, and the elderly patient with co-morbidities and polypharmacy on the other hand. We further suggest a method of comparing and choosing between the different hypoglycemic drugs available. Drugs should be considered not only based on their hypoglycemic effect but also on several other attributes such as effects on weight, glycemic durability, cardiovascular protection, individual experience with the drug, method of delivery and side effect profiles. Scoring the different attributes allows us to compare between different preparations and choose the most suitable drugs for each individual patient. Using our newly suggested system, a physician will first calculate the adequate HbA1c goal for his patient and then choose the drug that will best suit him, thus tailoring the treatment to the patients needs. PMID- 19557294 TI - Inflammation in Diabetic Encephalopathy is Prevented by C-Peptide. AB - Encephalopathy is an increasingly recognized complication of type 1 diabetes. The underlying mechanisms are not well understood, although insulin deficiency has been implicated. The spontaneously diabetic BB/Wor-rat develops neuro-behavioral deficits and neuronal cell death in hippocampus and frontal cortex, which can be prevented by insulinomimetic C-peptide. Here we examined whether contributing factors such as activation of innate immune mediators are responsive to C-peptide replacement. Seven-month diabetic BB/Wor-rats and those treated with full C peptide replacement were compared to age-matched control rats. Hippocampi of diabetic rats showed upregulation of RAGE and NF-kappaB, the former being localized to proliferating astrocytes. These changes were associated with increased expression of TNF-alpha, IL-1beta, IL-2 and IL-6 in hippocampi of diabetic rats. Full C-peptide replacement, which did not induce hyperglycemia, resulted in significant prevention of upregulation of RAGE expression, activation of NF-kappaB and activation of pro-inflammatory factors. In conclusion, impaired insulin activity is associated with upregulation of RAGE and pro-inflammatory factors, and these are likely to contribute to previously described oxidative and apoptotic neuronal cell death. Replacement of insulinomimetic C-peptide significantly prevents this cascade of events. PMID- 19557295 TI - GAD65-Specific Cytotoxic T Lymphocytes Mediate Beta-Cell Death and Loss of Function. AB - Autoimmunity to islet cell antigens like glutamic acid decarboxylase 65kD (GAD65) is associated with the destruction of insulin-producing beta-cells and progression to type 1 diabetes (T1D) in NOD mice and humans. T cell responses to GAD65 are detectable in the spleen of prediabetic NOD mice and in the peripheral blood of humans prior to the onset of overt hyperglycemia. Previous findings from our lab revealed that GAD65(546-554)-specific cytotoxic T lymphocytes (CTL) are present in naive NOD mice and are able to induce islet inflammation upon adoptive transfer into NOD.scid recipients. Additionally, we found that professional antigen-presenting cells (APC) generate the p546-554 epitope from a soluble GAD65 fragment, p530-554, and from GAD65 released by injured beta-cells in vivo. Here, we report that the GAD65 fragment p546-554 is a dominant CTL-inducing epitope which is naturally processed and presented by a GAD65-expressing beta-cell line. Further, co-culture of GAD65(546-554)-specific CTL with the beta-cells leads to a reduction in insulin production and the induction of perforin-mediated cell death. Collectively, these findings support a role for the cross-presentation of GAD65 antigen in the priming and enhancement of dominant GAD65-specific CTL responses, which can directly target beta-cells that display GAD65 epitopes. PMID- 19557293 TI - Adenosine Monophosphate-Activated Protein Kinase (AMPK) as a New Target for Antidiabetic Drugs: A Review on Metabolic, Pharmacological and Chemical Considerations. AB - In view of the epidemic nature of type 2 diabetes and the substantial rate of failure of current oral antidiabetic drugs the quest for new therapeutics is intensive. The adenosine monophosphate-activated protein kinase (AMPK) is an important regulatory protein for cellular energy balance and is considered a master switch of glucose and lipid metabolism in various organs, especially in skeletal muscle and liver. In skeletal muscles, AMPK stimulates glucose transport and fatty acid oxidation. In the liver, it augments fatty acid oxidation and decreases glucose output, cholesterol and triglyceride synthesis. These metabolic effects induced by AMPK are associated with lowering blood glucose levels in hyperglycemic individuals. Two classes of oral antihyperglycemic drugs (biguanidines and thiazolidinediones) have been shown to exert some of their therapeutic effects by directly or indirectly activating AMPK. However, side effects and an acquired resistance to these drugs emphasize the need for the development of novel and efficacious AMPK activators. We have recently discovered a new class of hydrophobic D-xylose derivatives that activates AMPK in skeletal muscles in a non insulin-dependent manner. One of these derivatives (2,4;3,5 dibenzylidene-D-xylose-diethyl-dithioacetal) stimulates the rate of hexose transport in skeletal muscle cells by increasing the abundance of glucose transporter-4 (GLUT-4) in the plasma membrane through activation of AMPK. This compound reduces blood glucose levels in diabetic mice and therefore offers a novel strategy of therapeutic intervention strategy in type 2 diabetes. The present review describes various classes of chemically-related compounds that activate AMPK by direct or indirect interactions and discusses their potential for candidate antihyperglycemic drug development. PMID- 19557296 TI - Factors Associated with the Prevalence of Diabetes Mellitus Among Elderly Men and Women Living in Mediterranean Islands: The MEDIS Study. AB - BACKGROUND: The aim of the present work was to evaluate the relationships between socio-demographic, clinical, lifestyle and psychological characteristics and the presence of diabetes mellitus, among elderly individuals without known cardiovascular disease. METHODS: During 2005-2007, 1190 elderly (aged 65 to 100 years) men and women (from Cyprus, Mitilini, Samothraki, Cephalonia, Crete, Lemnos, Corfu and Zakynthos) were enrolled. Socio-demographic, clinical and lifestyle factors were assessed using standard procedures. Diabetes mellitus was defined as fasting blood glucose >125 mg/dl or use of special medication. RESULTS: 21% of males and 23% of females had diabetes. Only 70% of diabetic participants were on a special diet and 76% were receiving pharmaceutical treatment. Diabetic individuals had higher prevalence of hypertension (80% vs. 64%, p < 0.001) and hypercholesterolemia (63% vs. 51%, p < 0.001) and reported lower physical activity status (p < 0.001), compared with non-diabetic participants. After adjusting for various confounders, hypertension and hypercholesterolemia were associated with a 144% (95% CI, 1.37-4.35) and 83% (95% CI, 1.13-2.94) higher likelihood of having diabetes, while moderate and vigorous exercise correlated with a 82% (95% CI, 0.09-0.81) and 67% (95% CI, 0.11-0.97) lower likelihood of diabetes. CONCLUSIONS: A considerable proportion of our elderly sample had diabetes and other metabolic disorders, almost 25% of which were untreated. Promotion of physical activities, even in the elderly, may contribute to reducing their burden of diabetes and provide them with a better quality of living. PMID- 19557297 TI - Effect of zinc supplementation on serum homocysteine in type 2 diabetic patients with microalbuminuria. AB - OBJECTIVES: Elevated homocysteine levels are considered to be an independent risk factor for cardiovascular complications in diabetic patients. The aim of this study was to find out if zinc supplementation improves homocysteine levels, which may exert vascular-protective effects in type 2 diabetes subjects with microalbuminuria. METHODS: In a randomized, double-blind, controlled, crossover study, 50 type 2 diabetic patients with microalbuminuria were subdivided into two groups and supplemented with 30 mg/d of zinc (group 1) or placebo (group 2) for three months with a four-week wash out period. Serum creatinine, vitamin B(12), folate, fasting plasma glucose, HbA1c, lipid profiles, zinc, homocysteine levels and random urine albumin were measured before and after the first and second phase of the study in all participants. RESULTS: Mean serum zinc was significantly increased after zinc supplementation (from 76 +/- 16 mug/dl to 93 +/- 20 microg/dl; p < 0.05), while there was no change in the placebo group (75 +/- 16 microg/dl to 75 +/- 15 microg/dl). With zinc supplementation, homocysteine levels reduced significantly (from 13.71 +/- 3.84 mumol/l to 11.79 +/- 3.06 mumol/l; p < 0.05), which did not occur on placebo (from 12.59 +/- 2.13 mumol/l to 13.36 +/- 2.03 mumol/l). Simple regression was used to show a positive correlation between urine albumin excretion and serum homocysteine (r = 0.37, p = 0.023). Vitamin B(12) and folate levels increased significantly in patients who received zinc in comparison to those who received placebo. A negative correlation was observed between homocysteine and vitamin B(12) concentration (r = -0.36, p = 0.025). CONCLUSION: Zinc supplementation reduced serum homocysteine and increased vitamin B(12) and folate concentrations in type 2 diabetic patients with microalbuminuria. PMID- 19557298 TI - Effects of elevated atmospheric CO2 and O3 on Hill activity and Ca2+/Mg2+-ATPase activity of Pinus tabulaeformis Carr. AB - The main photo-physiological characteristics of Pinus tabulaeformis Carr. were analyzed in open-top chambers under elevated carbon dioxide and ozone concentrations. The results indicated that the leaves net photosynthetic rates (p < 0.05), Hill activity, Ca2+/Mg2+-ATPase activity, soluble sugar and starch contents all increased under elevated carbon dioxide concentration in whole growing season. While under elevated ozone concentration, the leaves net photosynthetic rates, Hill activity, Ca2+/Mg2+-ATPase activity, soluble sugar and starch contents all decreased. Under elevated carbon dioxide and ozone concentration, the leaves net photosynthetic rates, Hill activity, soluble sugar and starch contents all increased, but Ca2+-ATPase activity increased during the earlier growing season, decreased in later growing season, while Mg2+-ATPase activity responded contrarily. PMID- 19557299 TI - Effects of a novel allelochemical ethyl 2-methyl acetoacetate (EMA) on the ultrastructure and pigment composition of cyanobacterium Microcystis aeruginosa. AB - Allelochemical ethyl 2-methyl acetoacetate (EMA) can significantly inhibit the growth of bloom-forming Microcystis aeruginosa. In order to assess the implication of the damage of EMA on the algal photosynthetic apparatus, the effects of EMA on the algal ultrastructure and pigment composition were investigated. At initial exposure time to EMA (0-40 h), algal allophycocyanin, phycoerythrin and carotenoid degraded firstly; chlorophyll a increased, especially by 47% in the algae exposed to 2 mg L(-1) of EMA; phycocyanin was not significantly affected; lipid bodies increased remarkably. After 40 h of EMA exposure, chlorophyll a decreased gradually, especially by 45% in the algae exposed to 4 mg L(-1) of EMA; lipid bodies greatly reduced but cyanophycin granules accumulated; thylakoid structures were dissolved or disappeared with the presence of numerous vacuoles. These results showed that all ophycocyanin, phycoerythrin and carotenoid were more sensitive to EMA than other pigments, the cells of M. aeruginosa was stressed by EMA with the occurrence of cyanophycin granules and the photosynthesis pigments and ultrastructure of M. aeruginosa were quickly destroyed by EMA with exposure time increasing. PMID- 19557300 TI - Measurement of detrusor wall thickness in women with overactive bladder by transvaginal and transabdominal sonography. AB - INTRODUCTION AND HYPOTHESIS: Women with overactive bladder (OAB) might have a greater detrusor wall thickness (DWT) suggestive of detrusor overactivity (DO). METHODS: DWT was measured by transabdominal ultrasonography (TAU) and transvaginal ultrasonography (TVU) in normal and women with OAB-dry and OAB-wet. The subjects were further classified as normal, hypersensitive bladder, or DO by urodynamic results. DWT measured by TVU at empty bladder, and TAU at 250 ml and bladder capacity were compared among symptom and urodynamic subgroups. RESULTS: TVU-measured DWT was significantly greater at the bladder neck than other sites of the bladder wall. No significant difference of TVU-measured DWT was noted among subgroups. No significant difference of TAU-measured DWT among subgroups at 250 ml, but DWT at bladder capacity was significantly greater in OAB-wet or DO than other subgroups. CONCLUSIONS: A greater DWT at bladder capacity measured by TAU can be useful as biomarker for DO in patients with OAB. PMID- 19557301 TI - Polish version of the Hypomania Checklist (HCL-32) scale: the results in treatment-resistant depression. AB - We performed the factor analysis of the Polish version of the Hypomania Check List (HCL-32) scale and assessed the utility of HCL-32 in discriminating patients with treatment-resistant and treatment non-resistant depression. The study included 1,051 patients with single or recurrent depressive episode among which 569 met the criteria for treatment-resistant depression. The Polish version of HCL-32 was employed to all patients. The Cronbach's alpha for entire scale was 0.93 which indicates high degree of consistency. The factor analysis of the scale yielded three factors with item loadings of 0.4 or more. Factor 1, comprising ten items connected with elevated mood and increased activity explained more than half of total variance, Factor 2 (two items) was connected with sexual activity, and factor 3 (three items) with irritability. The mean score of HCL-32 was significantly higher in treatment-resistant versus non-resistant depression (11.9 +/- 8.3 vs. 8.5 +/- 7.7, respectively, P < 0.001). Also, the percentage of patients having positive response to 14 or more items of the scale was significantly higher in treatment-resistant than in non-resistant depression (43.9 vs. 30.0%, respectively, P < 0.001). Therefore, using Polish version of HCL 32 we have confirmed the association between bipolarity and worse response to antidepressant drugs in patients with mood disorders. PMID- 19557302 TI - Effectiveness of IkappaB kinase inhibitors in murine colitis-associated tumorigenesis. AB - PURPOSE: Nuclear factor kappaB (NF-kappaB) activation is involved in various inflammatory illnesses, for example inflammatory bowel disease, and is thought to be a key factor in the tumor-promoting mechanism of inflammation-associated tumorigenesis. This study investigated whether inhibitors of IKKbeta, which is a critical kinase for NF-kappaB activation, reduce colitis-associated tumorigenesis in mice. METHODS: We used a mouse model of the disease whereby administration of azoxymethane (AOM) followed by repeated dextran sulfate sodium (DSS) ingestion causes severe colonic inflammation and the subsequent development of multiple tumors. Effects of IKKbeta inhibitors, NBD peptide, and IMD-0354 were examined. RESULTS: In a colitis-associated cancer model, treatment with the IKKbeta inhibitors NBD peptide and IMD-0354 significantly reduced the number of tumors compared with the untreated group. The tumors were also significantly smaller in the inhibitor-treated mice than in the untreated mice. Macrophage and neutrophil infiltration decreased with the inhibitor treatment. NF-kappaB activation and the expression of Cox-2 and iNOS were observed in tumor tissues, and the inhibitors ameliorated their expression. These inhibitors blocked NF-kappaB activation and the expression of proinflammatory cytokines mediated by the culture supernatant of inflamed colon in murine primary macrophages. In-vitro and in-vivo experiments showed that these drugs, especially NBD peptide, could also inhibit the proliferation of colonic epithelial cells. CONCLUSION: These results imply that IKKbeta-targeted NF-kappaB blockade is an attractive therapeutic approach for the prevention of colitis-associated tumors. PMID- 19557303 TI - Adverse allergic reaction to 131I MIBG. AB - No adverse allergic reactions to iodine-131-metaiodobenzylguanidine ((131)I MIBG) at a diagnostic dose have been reported in the English literature. This report of a skin eruption in a 35-year-old man after an intravenous injection of (131)I MIBG strongly suggests an adverse allergic reaction, and is the first to address such a side effect of (131)I MIBG at a diagnostic dose. Erythematous maculopapular eruptions, some of which were contiguous, were seen in a symmetric disposition on the patient's chest walls, elbows, neck and face 18 h after the (131)I MIBG injection. Antiallergic treatment resolved the lesions completely. There were no possible causes of the exanthema other than the (131)I MIBG injection. Urticaria related to the (131)I MIBG injection and caused by type I allergic reaction was suspected, and these findings point to the possible risk of a hitherto unreported allergic skin reaction to (131)I MIBG. We would like to draw the attention of nuclear physicians to this possible drawback of (131)I MIBG. PMID- 19557304 TI - Factor XIII deficiency as a potential cause of supratentorial haemorrhage after posterior fossa surgery. AB - BACKGROUND: Postoperative intracranial haemorrhage can be a dramatic event, carrying significant morbidity and mortality. Bleeding at sites remote from the operation area represents a small percentage of haemorrhages whose aetiology remains unclear (Harders et al. Acta Neurochir (Wien) 74(1-2):57-60, 1985). AIM: We present the case of a 60-year-old patient who underwent posterior fossa craniotomy for the removal of a space-occupying lesion and suffered supratentorial haemorrhage soon after the operation. RESULTS: A thorough postoperative investigation revealed low levels of factor XIII (FXIII), the factor mainly responsible for fibrin clot stabilisation. CONCLUSION: We suggest that reduced FXIII activity may be an important but preventable predisposing factor to remote postoperative haemorrhage in neurosurgical patients. PMID- 19557305 TI - Hemostatic and hemorrhagic problems in neurosurgical patients. AB - BACKGROUND: Abnormalities of the hemostasis can lead to hemorrhage, and on the other hand to thrombosis. Intracranial neoplasms, complex surgical procedures, and head injury have a specific impact on coagulation and fibrinolysis. Moreover, the number of neurosurgical patients on medication (which interferes with platelet function and/or the coagulation systems) has increased over the past years. METHOD: The objective of this review is to recall common hemostatic disorders in neurosurgical patients on the basis of the "new concept of hemostasis". Therefore the pertinent literature was searched to provide a structured and up to date manuscript about hemostasis in Neurosurgery. FINDINGS: According to recent scientific publications abnormalities of the coagulation system are discussed. Pathophysiological background and the rational for specific (cost)-effective perioperative hemostatic therapy is provided. CONCLUSIONS: Perturbations of hemostasis can be multifactorial and maybe encountered in the daily practice of neurosurgery. Early diagnosis and specific treatment is the prerequisite for successful treatment and good patients outcome. PMID- 19557306 TI - Anti-tumor effects of nitrosylcobalamin against spontaneous tumors in dogs. AB - PURPOSE: Given the limited options available to treat canine cancers, the use of companion animals for evaluating new drugs may identify better therapies for veterinary and human oncology. The anti-tumor effects of nitrosylcobalamin (NO Cbl), an apoptosis-inducing, vitamin B12-based carrier of nitric oxide (NO), was evaluated in four dogs with spontaneous cancer. EXPERIMENTAL DESIGN: (1) A 13 year-old female spayed Giant Schnauzer with inoperable thyroid carcinoma and hypercalcemia. (2) A 6 year-old male neutered Golden Retriever with a malignant peripheral nerve sheath tumor (MPNST). (3) A ten yr-old neutered male Bichon Frise with apocrine gland anal sac adenocarcinoma (AGACA). (4) A 7 year-old female spayed Labrador mix with spinal meningioma following partial surgical resection. Tumor regression was measured by physical exam and verified using ultrasound (case 1) and MRI (case 2-4). Serum chemistries and hematologic parameters were monitored throughout the studies. RESULTS: (1) The Giant Schnauzer demonstrated a 77% reduction in tumor volume after ten weeks of daily NO-Cbl treatment. (2) The Golden Retriever demonstrated a 53% reduction in tumor volume after 15 months of daily NO-Cbl therapy. (3) The Bichon Frise demonstrated a 43% regression of the primary tumor and a 90% regression of an iliac lymph node measured by MRI after 15 months of treatment. After 61 months, the dog currently has stable disease, normal liver enzymes, CBC analysis, and no evidence of toxicity. (4) The Labrador demonstrated complete regression of the residual tumor after 6 months of treatment. CONCLUSION: We have shown previously that NO-Cbl is endocytosed by malignant cells, resulting in intra-tumoral NO release. In this study, we have shown that daily long-term use of NO-Cbl induced responses in all dogs without any signs of toxicity. The use of NO-Cbl capitalizes on the tumor specific properties of the vitamin B12 receptor and represents a promising anti cancer therapy. PMID- 19557307 TI - Enhanced bacterial expression of several mammalian cytochrome P450s by codon optimization and chaperone coexpression. AB - To elucidate the effects of codon optimization and chaperone coexpression on the heterologous expression of mammalian cytochrome P450s (P450) in Escherichia coli, the expression of P450s 2B1, 2S1, 2U1, 2W1, and 27C1 were investigated. With codon optimization for N-terminus or the entire gene, the expression levels of P450 27C1, 2U1 and 2W1 increased 22-fold, 3.6-fold and 2.1-fold, respectively, while those for P450s 2B1 and 2S1 remained unchanged. With coexpression of E. coli molecular chaperones GroEL/ES, the expression level increased up to 14-fold for P450 27C1, and 3- to 5-fold for P450s 2B1, 2S1, and 2W1. Simultaneous application of these two techniques resulted in synergetic effects. PMID- 19557308 TI - Altered composition of Ralstonia eutropha poly(hydroxyalkanoate) through expression of PHA synthase from Allochromatium vinosum ATCC 35206. AB - The class III poly(hydroxyalkanoate) synthase (PHAS) genes (phaC and phaE) of a photosynthetic bacterium, Allochromatium vinosum ATCC 35206, were cloned, sequenced and expressed in a heterologous host. PCR coupled with a chromosomal gene-walking method was used to clone and subsequently sequence the contiguous phaC (1,068 bps) and phaE (1,065 bps) genes of A. vinosum ATCC 35206. BLASTP search of protein databases showed that the gene-products of phaC and phaE are different (<66% identities) from the previously reported class III PHASs such as those of A. vinosum DSM180. Domain analysis revealed the presence of a conserved alpha/beta-hydrolase fold in PhaC, the putative gene-product of phaC. Upon electroporation of a poly(hydroxybutanoate) (PHB)-negative mutant of Ralstonia eutropha PHB(-)4 with a shuttle plasmid pBHR1 containing the newly cloned phaC and phaE genes, the bacteria resumed the synthesis of PHB, albeit at a low level (4-5% of the cell dry wt) due to kanamycin selection pressure. We further showed that the recombinant strain grown in kanamycin-containing culture medium synthesized a blend of PHA that also contains a high content of 3 hydroxyoctanoate and 3-hydroxydecanoate as its repeat-unit monomers. Genomic analysis suggested the existence of two PHA synthase genes in R. eutropha. The results of this study not only make available a phylogenetically diverse type III phaC and phaE genes, but also confirm through kanamycin selection pressure the existence of multiple PHA biosynthesis systems in R. eutropha. PMID- 19557309 TI - Selective inhibition of the cyanobacterium, Microcystis, by a Streptomyces sp. AB - A Streptomyces strain, NT0401, was isolated from soil that selectively inhibited Microcystis strains but did not affect other microorganisms. Based on its morphology, physiology and 16S rDNA sequence, it was identified as Streptomyces grisovariabilis. The active substance produced by NT0401 was a water-soluble compound with a Mr <1 kDa that was stable over a broad pH range and at 100 degrees C for 20 min. This organism should be a potential environment-friendly strain for control of Microcystis blooms. PMID- 19557310 TI - Enhanced nitrogen removal in bio-electrochemical systems by pH control. AB - Microbial fuel cells can be designed to remove nitrogenous compounds out of wastewater, but their performance is at present limited to 0.33 kg NO(3) (-)-Nm( 3) net cathode compartment (NCC) d(-1). By maintaining the pH in the cathode at 7.2, nitrogen removal was increased from 0.22 to 0.50 kg NO(3) (-)-Nm(-3) NCC d( 1). Bio-electrochemical active microorganisms seem to struggle with the deterioration of their own environment due to slow proton fluxes. Therefore, the results suggest that an appropriate pH adjustment strategy is necessary to allow a sustained and enhanced biological activity in bio-electrochemical systems. PMID- 19557312 TI - The immunosuppressive activities of newly synthesized azaphenothiazines in human and mouse models. AB - In this study, we evaluated the activities of new types of azaphenothiazines in the following immunological assays: the proliferative response of human peripheral blood mononuclear cells induced by phytohemagglutin A or anti-CD3 antibodies; lipopolysaccharide-induced cytokine production by human PBMC; the secondary, humoral immune response in mice to sheep erythrocytes (in vitro); and delayed-type hypersensitivity in mice to ovalbumin (in vivo). In some tests, chlorpromazine served as a reference drug. The compounds exhibited differential inhibitory activities in the proliferation tests, with 10H-2,7-diazaphenothiazine (compound 1) and 6-(3-dimethylaminopropyl)diquinothiazine (compound 8) being most suppressive. Compound 1 was selected for further studies, and was found to be strongly suppressive in the humoral immune response even at low concentrations (1 microg/ml). Compound 1 also inhibited the delayed-type hypersensitivity lipopolysaccharide-induced production of tumor necrosis factor and interleukin-6 in cultures of human blood cells. As there were only two subjects in this study, the effects of these compounds on human blood cells need to be confirmed. In this paper, we also discuss the structure-activity relationships of selected compounds. PMID- 19557313 TI - Arginine methylation analysis of the splicing-associated SR protein SFRS9/SRP30C. AB - The human SFRS9/SRp30c belongs to the SR family of splicing regulators. Despite evidence that members of this protein family may be targeted by arginine methylation, this has yet to be experimentally addressed. In this study, we found that SFRS9 is a target for PRMT1-mediated arginine methylation in vitro, and that it is immunoprecipitated from HEK-293 lysates by antibodies that recognize both mono- and dimethylated arginines. We further observed that upon treatment with the methylation inhibitor Adox, the fluorescent EGFP-SFRS9 re-localizes to dot like structures in the cell nucleus. In subsequent confocal analyses, we found that EGFP-SFRS9 localizes to nucleoli in Adox-treated cells. Our findings indicate the importance of arginine methylation for the subnuclear localization of SFRS9. PMID- 19557314 TI - Dietary zinc glycine chelate on growth performance, tissue mineral concentrations, and serum enzyme activity in weanling piglets. AB - One hundred twenty crossbred piglets (Duroc x Landrace x Yorkshire) were used to determine the effects of dietary zinc glycine chelate on growth performance, tissue mineral concentrations, and serum enzyme activity. All pigs were allotted to four treatments and fed with basal diets supplemented with 0, 50, and 100 mg/kg Zn as zinc glycine chelate or 3,000 mg/kg Zn as zinc oxide (ZnO). After the 35-day feeding trial, results of the study showed that, compared to the control, average daily gain was improved (P < 0.05) for pigs fed 100 mg/kg Zn from zinc glycine chelate or 3,000 mg/kg Zn from ZnO and Zn concentrations in serum and M. longissimus dorsi were significantly enhanced by 100 mg/kg dietary zinc glycine chelate and 3,000 mg/kg ZnO. In addition, supplementation of 100 mg/kg zinc glycine chelate decreased (P < 0.05) the liver Fe level, liver Zn level, spleen Cu level, and kidney Cu level compared to that of the 3,000-mg/kg ZnO group. For feces mineral excretion, 3,000 mg/kg Zn from ZnO greatly increased the concentration of fecal Zn (P < 0.01) and Mn (P < 0.05) compared to that of the control or the 100-mg/kg zinc glycine chelate group. Moreover, alkaline phosphatase and Cu/Zn superoxide dismutase activities of pigs in 100 mg/kg zinc glycine chelate and ZnO treatments were greatly higher than that of the control. The results of present study showed that supplementation with zinc glycine chelate could improve growth and serum enzyme activities and could also decrease zinc excretion in feces in weanling pig compared to high dietary ZnO. PMID- 19557311 TI - Interdisciplinary review for correlation between the plant origin capsaicinoids, non-steroidal antiinflammatory drugs, gastrointestinal mucosal damage and prevention in animals and human beings. AB - BACKGROUND: The plant origin capsaicinoids (capsaicin, dihydrocapsaicin, norcapsaicin, dihydrocapsaicin, homocapsaicin, homodihydrocapsaicin) are well known and used as nutritional additive agents in the every day nutritional practice from the last 9,500 years; however, we had have a very little scientifically based knowledge on their chemistry, physiology and pharmacology in animal observations, and in humans up to the mid-twentieth century. Our knowledge about their chemistry, physiology, pharmacology entered to be scientifically based evidence from the year 1980, dominantly in animal observations. The human observations with capsaicin (capsaicinoids), in terms of good clinical practice, have been started only in the last 10-year period (from 1997) in randomized, prospective, multiclinical studies. The name of "capsaicin" used only in the physiological and pharmacological research both in animal experiments and in human observation. The "capsaicin" (as a "chemically" used natural compound) modifies the so-called capsaicin-sensitive afferent nerves depending on their applied doses. AIMS: The specific action of capsaicin (capsaicinoids) on sensory afferent nerves modifying gastrointestinal (GI) function (under very specific conditions) offers a possibility for the production of an orally applicable drug or for other drug combinations, which can be used in the human medical therapy. The production of new drug is based on the critical interdisciplinary review of the results obtained with capsaicinoids. MATERIALS AND METHODS: This paper gives an interdisciplinary and critical overview on the chemical, physiological, pharmacological and toxicological actions of the natural origin capsaicinoids (from the point of drug production) under conditions of acute, subacute and chronic administration in animal experiments and human observations, toxicology, pharmacokinetics). This interdisciplinary review covers the following main chapters: (1) physiological and pharmacological research tool by capsaicin in the animals and human beings, (2) capsaicin research in animals (including the acute, subacute toxicology and chronic toxicology metabolism, genotoxicology), (3) capsaicin observation with capsaicin in human beings. CONCLUSION: (1) The capsaicin used in the physiological and pharmacological observations (in animals and human beings) chemically represents different chemical compounds, which can be obtained from the plants (paprika, chilli, etc.), (2) capsaicinoids are able to modify the capsaicin-sensitive afferent nerves, which have principle roles in the defence of different organs (including the gastrointestinal tract [against the different chemicals, heat, strech, chemical millieu-induced damage], (3) the application of capsaicin (capsaicinoids) can be repeated for the beneficial effects on the gastrointestinal tract as those in animal experiments. After this interdisciplinary and critical review, this paper demonstrates the well-planned research pathways of the discoveries of capsaicinoids from plant chemistry, via physiology, pharmacology and toxicology in animal experiments and human observations. PMID- 19557315 TI - Intravenous immunoglobulin in lupus panniculitis. AB - Systemic lupus erythematosus (SLE) is a disease of unknown cause that may involve one or many organ or systems. Skin involvement is a major feature in this disease, and a wide variety of skin conditions may be present. Lupus erythematosus panniculitis (LEP) constitutes a rare form of cutaneous lupus characterized by recurrent nodular or plaque lesions that can vary from a benign and mild course to a more disfiguring disease. Initial therapy includes corticosteroids, antimalarials, and azathioprine and, in refractory cases, two antimalarials in association, mycophenolate mofetil, or other immunomodulators. Intravenous immuglobulin (IVIG) is used in many autoimmune disorders, like in SLE, although clinical trials have not yet taken place. In this report, we review skin manifestations of SLE and their treatment, IVIG, and finally a case of LEP successfully treated with IVIG when other therapy modalities failed. PMID- 19557316 TI - The pathogenesis of neural injury in animal models of the antiphospholipid syndrome. AB - Circulating antiphospholipid antibodies (aPL) are associated with central nervous system dysfunction in antiphospholipid syndrome (APS) patients and in a mouse model of APS. We propose a logical pathway of how experimental APS (eAPS) causes brain dysfunction: binding of the antibodies to the brain endothelium evoking microthrombosis, endothelial dysfunction, and IgG leakage through the blood-brain barrier (BBB), then secondary inflammatory cell spread around blood vessels and production of cytokines by these inflammatory cells leading to further disruption of the BBB. The diffuse brain endothelial dysfunction would result in extravasation of serum proteins including APS IgG and activated thrombin, which may induce the behavioral changes observed in the APS mice. We have collected data from the mouse eAPS model which supports this hypothesis. Elucidating the mechanism of the pathogenicity of aPL in vitro and in vivo will serve as a much needed basis for developing new therapeutic modalities in this important disorder. PMID- 19557317 TI - High-dose intravenous immunoglobulin (IVIG) therapy in autoimmune skin blistering diseases. AB - Treatment of autoimmune bullous skin diseases can often be challenging and primarily consists of systemic corticosteroids and a variety of immunosuppressants. Current treatment strategies are effective in most cases but hampered by the side effects of long-term immunosuppressive treatment. Intravenous immunoglobulin (IVIG) is one potential promising therapy for patients with autoimmune bullous skin diseases, and evidence of its effectiveness and safety is increasing. A number of autoimmune bullous skin diseases have been identified in which IVIG treatment may be beneficial. However, experience with IVIG in patients with autoimmune skin blistering disease is limited, where it is recommended for patients not responding to conventional therapy. The mode of action of IVIG in autoimmune diseases, including bullous diseases is far from being completely understood. We here summarize the clinical evidence supporting the notion, that IVIG is a promising therapeutic agent for the treatment of patients with autoimmune bullous skin disease. In addition, we review the proposed modes of action. In the future, randomized controlled trials are necessary to better determine the efficacy and adverse effects of IVIG in the treatment of autoimmune bullous skin diseases. In addition, insights into IVIG's mode of action might enable us to develop novel therapeutics to overcome the current shortage of IVIG. PMID- 19557319 TI - Lupus and pregnancy--15 years of experience in a tertiary center. AB - This retrospective study was designed to evaluate the outcome of pregnancies in women diagnosed with systemic lupus erythematosus (SLE) followed in a tertiary fetal-maternal center. Data were collected from clinical charts between January 1993 and December 2007, with a total of 136 pregnancies (107 patients). Mean maternal age was 29 years, with the vast majority of patients being Caucasian. Most patients were in remission 6 months prior to pregnancy (93%) and the most frequently affected organs were the skin and joints. Renal lupus accounted for 14% of all cases. Twenty-nine percent of patients were positive for at least one antiphospholid antibody (aPL) and nearly 50% had positive SSa/SSb antibodies. All patients with positive aPL received low-dosage aspirin and low-molecular-weight heparin (LMWH). There were no pregnancy complications in more than 50% of cases and hypertensive disease and intrauterine growth restriction were the most common adverse events. There were 125 live births, one neonatal death, eight miscarriages, and three medical terminations of pregnancy. Preterm delivery occurred in 25% of pregnancies. Our results are probably the conjoined result of a multidisciplinary approach together with a systematic management of SLE pregnancies, with most patients keeping their prior SLE medication combined with low-dosage aspirin and LMWH in the presence of aPL. PMID- 19557318 TI - Predictors of pregnancy outcome in antiphospholipid syndrome: a review. AB - In pregnant women, antiphospholipid syndrome (APS) is associated with an increased risk of preeclampsia, fetal intrauterine growth restriction, and other complications related to uteroplacental insufficiency. In the last two decades, several studies were performed to identify the predictive role of some parameters in relation to obstetric outcome in APS patients. Among these, the uterine velocimetry Doppler is the most studied. It provides a non-invasive method for the study of uteroplacental blood flow, being able to detect a condition of impaired placental perfusion, due to the presence of circulating antiphospholipid antibodies (aPL). To date, the uterine artery Doppler velocimetry resulted to be a useful tool to identify APS pregnancies at higher risk of adverse pregnancy outcome. False-positive IgM for toxoplasmosis, others, rubella, cytomegalovirus, herpes viruses (TORCH) complex is associated to a worse pregnancy outcome because it reflects a dysregulation of the immune system which may amplify placental autoimmune damage. Moreover low levels of complement components are related to an increased incidence of obstetrical complications, suggesting that placental deposition of immune complexes and activation of complement cascade may contribute to placental failure APS related. The abnormal uterine Doppler velocimetry, false-positive TORCH IgM and low levels of complement components can be considered prognostic indexes of poor pregnancy outcome in APS. PMID- 19557320 TI - Effect of decreased inspiratory times on tidal volume. Bench model simulating cardiopulmonary resuscitation. AB - BACKGROUND: During cardiopulmonary resuscitation (CPR) with a chest compression rate of 60-100/min the time for secure undisturbed ventilation in the chest decompression phase is only 0.3-0.5 s and it is unclear which tidal volumes could be delivered in such a short time. OBJECTIVES: Attempts were made to assess the tidal volumes that can be insufflated in such a short time window. METHODS: In a bench model tidal volumes were compared in simulated non-intubated and intubated patients employing an adult self-inflating bag-valve with inspiratory times of 0.25, 0.3, and 0.5 s. Respiratory system compliance values were 60 mL/cmH(2)O being representative for respiratory system conditions shortly after onset of cardiac arrest and 20 mL/cmH(2)O being representative for conditions after prolonged cardiac arrest. RESULTS: With a respiratory system compliance of 60 mL/cmH(2)O, tidal volumes (mean+/-SD) in non-intubated versus intubated patients were 144+/-13 mL versus 196+/-23 mL in 0.25 s (p<0.01), 178+/-10 versus 270+/-14 mL in 0.3 s (p<0.01), and 310+/-12 mL versus 466+/-20 mL in 0.5 s (p<0.01). With a respiratory system compliance of 20 mL/cmH(2)O, tidal volumes in non-intubated patient versus intubated patients were 128+/-10 mL versus 186+/-20 mL in 0.25 s (p<0.01), 158+/-17 versus 250+/-14 mL in 0.3 s (p<0.01) and 230+/-21 mL versus 395+/-20 mL in 0.5 s (p<0.01). CONCLUSIONS: Ventilation windows of 0.25, 0.3, and 0.5 s were too short to provide adequate tidal volumes in a simulated non intubated cardiac arrest patient. In a simulated intubated cardiac arrest patient, ventilation windows of at least 0.5 s were necessary to provide adequate tidal volumes. PMID- 19557321 TI - [Acromegaly-associated lesions of the nasal mucosa. Case report]. AB - Acromegaly is a rare disease caused by a growth-hormone-secreting pituitary adenoma. Symptoms include enlargement of the hands, feet, and jaw with growing dental interspaces, as well as hypertrophy of the tongue and nasal and sinusoidal mucosa. The two latter symptoms are mostly responsible for the accompanying obstructive sleep apnea syndrome. Besides these "cosmetic" symptoms, the disease is associated with hypertension and diabetes mellitus, as well as with an increased risk for adenomas and carcinomas of the colon. The average time span from first symptom to diagnosis is well over 6 years; a single determination of insulin-like growth factor 1 in serum can confirm the disease. The treatment of choice remains surgical resection of the adenoma in suitable patients, whereas in extensive disease with invasion of surrounding tissue, drug therapy and/or radiotherapy may be necessary. PMID- 19557322 TI - [Speech developmental delay in preschool children: evaluation of screening with anamnesis questions]. AB - BACKGROUND: In western societies, delay of speech development is frequent and important as it may affect the entire development of a child. Therefore, early diagnosis and treatment is of outstanding relevance. For screening purposes in clinical practice, questionnaires focussing on speech development are assumed to be both valid and economic assessment tools. In this study, results of a questionnaire based on "milestones in early childhood development" as previously reported by Michaelis for German children were compared with results of systematic German speech development tests. PATIENTS AND METHODS: In a retrospective study, data were collected from 25 German preschool children. Their parents had completed a questionnaire on their child's development prior to the examination with 4 questions focussing on speech development. In this report, the answers on these 4 questions are evaluated against the background of systematic German speech development tests, i.e. the H-SET and PLAKSS, and an unsystematic examination by an experienced speech and language phonologist. RESULTS: The questionnaire had a sensitivity of 50% and a specificity of 7%. The positive and negative predictive values were 55.9% and 5%, respectively. CONCLUSION: A questionnaire based on "milestones in early childhood development" as previously reported by Michaelis for German children, is clinically invalid for screening speech development in preschool children. PMID- 19557323 TI - [Complication rate of out-patient removal of ear wax: systematic review of the literature]. AB - BACKGROUND: Cerumen impaction is known as a frequent problem both in general practice as well as in community based ENT surgeries. The aim of the present review was to describe the frequency and types of complications due to different methods of ear wax removal. METHODS: A systematic review of the literature was carried out (Pubmed, Cochrane, Scopus) and data from the arbitration board for medical liability were evaluated. RESULTS: Controlled studies addressing the complications of ear wax removal are lacking in the literature and absolute risks could not be reported. Perforation of the tympanic membrane was observed in up to 1% of the cases. Syringing seems to result in a greater number of complications than mechanical removal using a curette. CONCLUSIONS: Complication rates were found to be higher than previously assumed. However, none of the publications had initially been intended to detect adverse events. It can be concluded that syringing can be delegated to trained staff members only if pre-existing defects of the tympanic membrane have been excluded. PMID- 19557325 TI - Detecting concealed knowledge using a novel attentional blink paradigm. AB - Traditional concealed information paradigms rely on the idea that stimuli that are meaningful to a person (critical items) will draw attentional resources disproportionately, relative to stimuli that are not (irrelevant items), generating detectable differences on a suitable dependent variable (behavioral, psychophysiological, or neural). Here, we introduce a behavioral paradigm that could be used to reveal concealed information by exploiting the link between concealed information and attentional processes more directly. This novel paradigm is based on the attentional blink phenomenon in which detection of a stimulus reduces detection accuracy rates of subsequent target stimuli within a 200-500 ms time window. We hypothesized that a well-known face used as a critical item could capture attentional resources automatically, making it harder to detect the occurrence of a subsequently presented target face. The results confirmed this hypothesis, and showed that concealed knowledge of a famous face could be detected in 9 out of 12 individuals by looking for a relative dip in target detection accuracy after the presentation of a critical item. PMID- 19557324 TI - 4-Hydroxy-2-nonenal induces mitochondrial dysfunction and aberrant axonal outgrowth in adult sensory neurons that mimics features of diabetic neuropathy. AB - Modification of proteins by 4-hydroxy-2-nonenal (4-HNE) has been proposed to cause neurotoxicity in a number of neurodegenerative diseases, including distal axonopathy in diabetic sensory neuropathy. We tested the hypothesis that exposure of cultured adult rat sensory neurons to 4-HNE would result in the formation of amino acid adducts on mitochondrial proteins and that this process would be associated with impaired mitochondrial function and axonal regeneration. In addition, we compared 4-HNE-induced axon pathology with that exhibited by neurons isolated from diabetic rats. Cultured adult rat dorsal root ganglion (DRG) sensory neurons were incubated with varying concentrations of 4-HNE. Cell survival, axonal morphology, and level of axon outgrowth were assessed. In addition, video microscopy of live cells, western blot, and immunofluorescent staining were utilized to detect protein adduct formation by 4-HNE and to localize actively respiring mitochondria. 4-HNE induced formation of protein adducts on cytoskeletal and mitochondrial proteins, and impaired axon regeneration by approximately 50% at 3 microM while having no effect on neuronal survival. 4-HNE initiated formation of aberrant axonal structures and caused the accumulation of mitochondria in these dystrophic structures. Neurons treated with 4-HNE exhibited a distal loss of active mitochondria. Finally, the distal axonopathy and the associated aberrant axonal structures generated by 4-HNE treatment mimicked axon pathology observed in DRG sensory neurons isolated from diabetic rats and replicated aspects of neurodegeneration observed in human diabetic sensory neuropathy. PMID- 19557326 TI - Glutathione exhibits antibacterial activity and increases tetracycline efficacy against Pseudomonas aeruginosa. AB - Glutathione (GSH) plays important roles in pulmonary diseases, and inhaled GSH therapy has been used to treat cystic fibrosis (CF) patients in clinical trials. The results in this report revealed that GSH altered the sensitivity of Pseudomonas aeruginosa to different antibiotics through pathways unrelated to the oxidative stress as generally perceived. In addition, GSH and its oxidized form inhibited the growth of P. aeruginosa. PMID- 19557328 TI - Posttranslational modulation on the biological activities of molecular chaperones. AB - Molecular chaperones are a family of proteins that were first noticed to exist about 45 years ago from their increased transcription under heat shock conditions. As a result, the regulation of their encoding genes has been subject to extensive studies. Recent studies revealed that the biological activities of molecular chaperones can also be effectively modulated at the protein level. The ways of modulation so far elucidated include allosteric effect, covalent modification, protein-protein interaction, and conformational alteration induced by such macro-environmental conditions as temperature and pH. These latter aspects were reviewed here. Emphasized here is the importance of such immediate structural alterations that lead to an immediate activity increase, providing the immediate protection needed for the cells to survive the stress conditions. PMID- 19557329 TI - Early-phase clinical trials of anti-HIV drugs-understanding and discussion. AB - Innovative anti-HIV drugs developed by local sponsors in China have come into the stage of early-phase clinical trials. How to systemically design the clinical trials of innovative anti-HIV drugs still remains a challenge for them. This article references the literature and the experience of reviewers, to introduce general considerations concerning early-phase clinical trials of innovative anti HIV drugs. PMID- 19557327 TI - Towards transgenic primates: What can we learn from mouse genetics? AB - Considering the great physiological and behavioral similarities with humans, monkeys represent the ideal models not only for the study of complex cognitive behavior but also for the preclinical research and development of novel therapeutics for treating human diseases. Various powerful genetic technologies initially developed for making mouse models are being explored for generating transgenic primate models. We review the latest genetic engineering technologies and discuss the potentials and limitations for systematic production of transgenic primates. PMID- 19557330 TI - The CXXC finger 5 protein is required for DNA damage-induced p53 activation. AB - The tumor suppressor p53 is a critical component of the DNA damage response pathway that induces a set of genes responsible for cell cycle arrest, senescence, apoptosis, and DNA repair. The ataxia telangiectasia mutated protein kinase (ATM) responds to DNA-damage stimuli and signals p53 stabilization and activation, thereby facilitating transactivation of p53 inducible genes and maintainence of genome integrity. In this study, we identified a CXXC zinc finger domain containing protein termed CF5 as a critical component in the DNA damage signaling pathway. CF5 induces p53 transcriptional activity and apoptosis in cells expressing wild type p53 but not in p53-deficient cells. Knockdown of CF5 inhibits DNA damage-induced p53 activation as well as cell cycle arrest. Furthermore, CF5 physically interacts with ATM and is required for DNA damage induced ATM phosphorylation but not its recruitment to chromatin. These findings suggest that CF5 plays a crucial role in ATM-p53 signaling in response to DNA damage. PMID- 19557331 TI - Crystal structure of the MH2 domain of Drosophila Mad. AB - The decapentaplegic(Dpp), a member of the TGF-beta superfamily, plays a pivotal role in the control of proliferation, global patterning and induction of specific cell fates during Drosophila development. Mother against Dpp(Mad) is the founding member of the conserved Smad protein family which specifically transduces the intracellular TGF-beta signaling cascade. Here we report the 2.80 A structure of the MH2 domain of Mad(Mad-MH2) that was readily superposed to the mammal Smad-MH2 structures. This unphosphorylated Mad-MH2 forms a symmetric homotrimer in crystals, consistent with the result of the size-exclusion chromatography that Mad-MH2 exhibited a propensity for concentration-dependent oligomerization prior to phosphorylation. Structural analysis revealed that the formation of homotrimeric Mad-MH2 is mainly mediated by contacts involving the extreme C terminal SSVS motif, and is strengthened by phosphorylation of the last two Ser residues which was confirmed by the gel filtration analysis of the pseudophosphorylated Mad-MH2(DVD). Intriguingly, the homotrimer within an asymmetric unit only possesses two ordered C-terminal tails, reminiscent of the arrangement of the R-Smad/Smad4 complexes, indicating that the subunit with a flexible SSXS motif would be readily replaced by Co-Smad to form a functional heterotrimer. PMID- 19557333 TI - Identification and functional analysis of a CDE/CHR element in the POLD1 promoter. AB - DNA polymerase delta is encoded by the POLD1 gene, the transcription of which is strictly cell cycle-dependent. However, the means by which POLD1 transcription is regulated by the cell cycle mechanism is currently unknown. We discovered a novel element in the POLD1 promoter known as a CDE(cell cycle-dependent element)/CHR(cell cycle gene homology region) element. A series of luciferase reporter constructs containing various POLD1 promoter mutations were used to investigate the role of the CDE/CHR element in POLD1 transcription. When the CDE/CHR element was mutated, the promoter activity was up-regulated, and the cell cycle related factors E2F1 and p21 stopped regulating the promoter. Furthermore, cell cycle-dependent changes in the promoter activity required the integrative CDE/CHR element. Electrophoretic mobility shift assay (EMSA) revealed the presence of at least three types of DNA/protein complexes binding to the CDE/CHR element. Our findings provide strong evidence that the CDE/CHR-like sequence is an active functional element in the POLD1 promoter, which is important for the cell cycle regulation of the POLD1 gene. PMID- 19557334 TI - Progress in the detection of human genome structural variations. AB - The emerging of high-throughput and high-resolution genomic technologies led to the detection of submicroscopic variants ranging from 1 kb to 3 Mb in the human genome. These variants include copy number variations (CNVs), inversions, insertions, deletions and other complex rearrangements of DNA sequences. This paper briefly reviews the commonly used technologies to discover both genomic structural variants and their potential influences. Particularly, we highlight the array-based, PCR-based and sequencing-based assays, including array-based comparative genomic hybridization (aCGH), representational oligonucleotide microarray analysis (ROMA), multiplex amplifiable probe hybridization (MAPH), multiplex ligation-dependent probe amplification (MLPA), paired-end mapping (PEM), and next-generation DNA sequencing technologies. Furthermore, we discuss the limitations and challenges of current assays and give advices on how to make the database of genomic variations more reliable. PMID- 19557332 TI - Differential transcription-activating capability of NS1 proteins from different influenza virus subtypes expressed in yeast. AB - Influenza A virus NS1 protein is an important regulatory factor with multiple functions and contributes greatly to viral pathogenesis. In the present study, transcription-activating potential of NS1 from different influenza A virus subtypes was examined in yeast two-hybrid system. The bait vectors containing different NS1 genes, along with an empty prey vector, were transformed into yeast AH109(for growth assay on QDO plate and alpha-galactosidase assay), and Y187(for beta-galactosidase assay). AH109 transformants with NS1 gene from H1N1, H5N1, and H9N2 viruses grew vigorously on the QDO plate and secreted high level of alpha galactosidase. Also, Y187 bearing the above NS1 genes exhibited enhanced beta galactosidase activity. Nevertheless, H3N2-NS1-transformed AH109 and Y187 yeasts did not grow on QDO plate and secrete beta-galactosidase, respectively. These findings denote the remarkable variation in NS1 proteins from different influenza A virus subtypes on the transcription-stimulating capability in yeast. PMID- 19557335 TI - DNA barcoding: species delimitation in tree peonies. AB - Delimitations of species are crucial for correct and precise identification of taxa. Unfortunately "species" is more a subjective than an objective concept in taxonomic practice due to difficulties in revealing patterns of infra- or inter specific variations. Molecular phylogenetic studies at the population level solve this problem and lay a sound foundation for DNA barcoding. In this paper we exemplify the necessity of adopting a phylogenetic concept of species in DNA barcoding for tree peonies (Paeonia sect. Moutan). We used 40 samples representing all known populations of rare and endangered species and several populations of widely distributed tree peonies. All currently recognized species and major variants have been included in this study. Four chloroplast gene fragments, i.e. ndhF, rps16-trnQ, trnL-F and trnS-G (a total of 5040 characters, 96 variable and 69 parsimony-informative characters) and one variable and single copy nuclear GPAT gene fragment (2093-2197 bp, 279 variable and 148 parsimony informative characters) were used to construct phylogenetic relationships among the taxa. The evolutionary lineages revealed by the nuclear gene and the chloroplast genes are inconsistent with the current circumscriptions of P. decomposita, P. jishanensis, P. qiui, and P. rockii based on morphology. The inconsistencies come from (1) significant chloroplast gene divergence but little nuclear GPAT gene divergence among population systems of P. decomposita + P. rockii, and (2) well-diverged nuclear GPAT gene but little chloroplast gene divergence between P. jishanensis and P. qiui. The incongruence of the phylogenies based on the chloroplast genes and the nuclear GPAT gene is probably due to the chloroplast capture event in evolutionary history, as no reproductive barriers exist to prevent inter-specific hybridization. We also evaluated the suitability of these genes for use as DNA barcodes for tree peonies. The variability of chloroplast genes among well-defined species or population systems of a species complex is 4.82 times the figure within the groups, and the GPAT gene is twice as variable between the groups as within the groups. The number of completely divergent sites is sufficient to mark the two subsections, the two species in subsection Delavayanae, and the well-divergent species in subsection Vaginatae. But the genes currently used either from the chloroplast genome or from the nuclear genome alone cannot correctly assign samples of P. decomposita, P. jishanensis, P. qiui, or P. rockii to the species as currently defined. We conclude that (1) DNA barcoding should be based on prior phylogenetic studies to understand the evolutionary lineages and how well the taxonomic species correspond to the lineages; (2) it is unlikely to find a single short fragment as a barcode for every plant and such a fragment could result in misidentification when a chloroplast capture event happened in the evolutionary history of plants like tree peonies; and (3) we suggest striving for a universal marker at the familial level and locally universal barcodes within a family instead of looking for a universal barcode for all plants. PMID- 19557336 TI - The influence of mechanical loading on osseointegration: an animal study. AB - Osseointegration of implant provides a stable support for the prosthesis under functional loads. The timing of loading is a critical parameter that can govern the success of the osseointegration of implant. However, it is not clear whether the early loading can affect the success of osseointegration, or whether the no loading healing period can be shortened. This paper presents an animal study conducted to investigate how external loads influence the osseointegration at the initial stage of healing. Titanium implants were inserted into the goat tibia laterally, and different axial loadings were applied to the implants in 4 weeks after surgery. After the 2 weeks period of early loading, animals were sacrificed and the tibia bones with the implants were cut off from the bodies. Then mechanical test was employed to find out the differences in the pull-out force, and shear strength at the bone-implant interface between the non-loaded and the loaded implants. The implant-bone interfaces were analyzed by histomorphometric method, SEM (scanning electron micrograph) and EDS (energy density spectrum). The results indicated that the bone-implant interface did not well integrate 4 weeks after surgery, and the fibrous tissue could be found at the interfaces of the specimens without loadings. While the results of loaded specimens with 10 N axial force showed that that parts of the interface were well integrated, indicating that the early mild loading may play a positive role in the process of the osseointegration. The results support that a certain range of external loading would influence the process of osseointegration, and appropriate mechanical loading can be applied to shorten the osseointegration period after surgery. PMID- 19557337 TI - Mechanisms of invasive population establishment and spread of pinewood nematodes in China. AB - This paper summarizes the results of our study of the pinewood nematode (Bursaphelenchus xylophilus). By population genetic analysis, it was determined that there was no genetic bottle caused by the founder effect and genetic drift in the Chinese invasive population. Multiple invasions with large amounts of nematodes from different sources led to rich genetic diversity in the invasive population. Keeping high genetic diversity in the invasive process may be one of the genetic mechanisms in its successful invasion. By testing interspecies competition, it was shown that, with high fecundity and a strong competitive ability, B. xylophilus outcompeted the native species B. mucronatus in the natural ecosystem during the invasion process. Competitive displacement may be one of the ecological mechanisms of B. xylophilus's invasion. In addition, an unequal interspecific hybridization with introgress was in favor of the invasive species which also accelerated the replacement of B. mucronatus by B. xylophilus. The structures, functions and evolutions of a few important genes that are closely related to the ecological adaptation of pinewood nematodes were studied to explore the molecular mechanism of its ecological adaptations. Further, the resistance and resilience mechanism of the pine ecosystem invaded by pinewood nematodes was also investigated. The results of these studies uncovered a portion of the genetic and ecological mechanisms of PWN's successful invasion and laid a foundation for further study to obtain a comprehensive interpretation of the mechanisms of the nematode invasion. The results also provided a scientific basis for effectively controlling the occurrence and spread of pine wilt disease which is caused by nematodes. Various aspects requiring further investigation are considered. PMID- 19557338 TI - The linkage between reverse gyrase and hyperthermophiles: a review of their invariable association. AB - With the discovery of reverse gyrase in 1972, from Yellowstone National Park, isolated from Sulfolobus acidocaldarius, it has been speculated as to why reverse gyrase can be found in all hyperthermophiles and just what exactly its role is in hyperthermophilic organisms. Hyperthermophiles have been defined as organisms with an optimal growth temperature of above 85 degrees C. Reverse gyrase is responsible for the introduction of positive supercoils into closed circular DNA. This review of reverse gyrase in hyperthermophilic microorganisms summarizes the last two decades of research performed on hyperthermophiles and reverse gyrase in an effort to provide an up to date synopsis of their invariable association. From the data gathered for this review it is reasonable to hypothesize that reverse gyrase is closely tied to hyperthermophilic life. PMID- 19557340 TI - Nematicidal activity of Paecilomyces spp. and isolation of a novel active compound. AB - Many species of Paecilomyces are entomogenous fungi and several are efficacious toward nematodes. To study the potential of Paecilomyces species in controlling nematodes, fungal extracts of 40 Paecilomyces spp. were evaluated for their nematicidal activity against Bursaphelenchus xylophilus and Panagrellus redivivus. The extracts of six Paecilomyces spp. exhibited the nematicidal activity against P. redivivus, and 11 species exhibited the nematicidal activity against B. xylophilus. The methanol extract of strain 1.01761 incubating on Czapek solid medium killed more than 95% P. redivivus in 24 h at 5 mg/ml, and the filtrate of strain 1.01788 cultured in Sabouraud's broth medium resulted in 90% mortality of B. xylophilus in 24 h at 5 mg/ml. A novel nematicidal compound 4-(4' carboxy-2'-ethyl-hydroxypentyl)-5,6-dihydro-6-methylcyclobuta[b]pyridine-3,6 dicarboxylic acid, was isolated from Paecilomyces sp. YMF1.01761. The LD(50) value of the compound within 24 h against P. redivivus was 50.86 mg/L, against Meloidogyne incognita was 47.1 mg/L, and against B. xylophilus was 167.7 mg/L. PMID- 19557339 TI - Microbial diversity of a sulfide black smoker in main endeavour hydrothermal vent field, Juan de Fuca Ridge. AB - Submarine hydrothermal vents are among the least-understood habitats on Earth but have been the intense focus of research in the past 30 years. An active hydrothermal sulfide chimney collected from the Dudley site in the Main Endeavour vent Field (MEF) of Juan de Fuca Ridge was investigated using mineralogical and molecular approaches. Mineral analysis indicated that the chimney was composed mainly of Fe-, Zn-and Cu-rich sulfides. According to phylogenetic analysis, within the Crenarchaeota, clones of the order Desulfurococcales predominated, comprising nearly 50% of archaeal clones. Euryarchaeota were composed mainly of clones belonging to Thermococcales and deep-sea hydrothermal vent Euryarchaeota (DHVE), each of which accounted for about 20% of all clones. Thermophilic or hyperthermophilic physiologies were common to the predominant archaeal groups. More than half of bacterial clones belonged to epsilon-Proteobacteria, which confirmed their prevalence in hydrothermal vent environments. Clones of Proteobacteria (gamma-, delta-, beta-), Cytophaga-Flavobacterium-Bacteroides (CFB) and Deinococcus-Thermus occurred as well. It was remarkable that methanogens and methanotrophs were not detected in our 16S rRNA gene library. Our results indicated that sulfur-related metabolism, which included sulfur-reducing activity carried out by thermophilic archaea and sulfur-oxidizing by mesophilic bacteria, was common and crucial to the vent ecosystem in Dudley hydrothermal site. PMID- 19557341 TI - Evaluation of three molecular methods of repetitive element loci for differentiation of Mycobacterium avium subsp. paratuberculosis (MAP). AB - The aim of the present study is to evaluate the efficiency of three methods to determine the molecular diversity of 34 Mycobacterium avium subsp. paratuberculosis (MAP) strains isolated from 17 cattle herds. The applied methods included the analysis of sequence polymorphism of the mononucleotide (G1 and G2) and trinucleotide sequences (GGT) of the Short Sequence Repeats (SSR) and the determination of size polymorphism of 9 different Mycobacterial Interspersed Repetitive Units (MIRU) and 6 Variable Number Tandem Repeats (VNTR). Sequence analysis of SSR of 34 isolates showed 4, 6, and 2 alleles of G1, G2, and GGT repeats, respectively. The amplification of the investigated 9 MIRU units revealed only two discriminatory genotyping systems (MIRU2 and MIRU3). Out of 6 VNTR PCR differentiation methods, only one method could be recommended for genotyping purposes. The profile 7g-12g-4ggt-II-b-2 of the combination systems G1 G2-GGT-MIRU2-MIRU3-VNTR1658 dominates among the examined isolates and was detected in 14.7% of the isolates. The use of certain repetitive loci of SSR, MIRU, and VNTR techniques in this study showed greater potential than others for the characterization of MAP isolates. The recommended loci can be used for the epidemiological tracing of MAP field strains and to determine the relationships between isolates in different herds. PMID- 19557342 TI - The three-dimensional morphology of Candida albicans as seen by high-resolution scanning electron microscopy. AB - The fine structure of Candida albicans has been repeatedly described by transmission electron microscopy, whereas studies by high-resolution scanning electron microscopy (HRSEM) are rare and devoted solely to the study of its external morphology. This report describes the results of an HRSEM study on C. albicans carried out by an osmium maceration protocol modified to better retain the structural characteristics of this yeast. Thus, we visualized various intracellular structures including invaginations of cell membrane (plasmalemmasomes), nuclear envelope, mitochondria, the vacuolar system, and two additional structures that might represent a form of endoplasmic reticulum and the Golgi apparatus. The present investigation, which for the first time shows the organelles of C. albicans at the 3D level, may lead to a better understanding of its cell physiology. PMID- 19557343 TI - A new Dactylella species from Orbilia alba. AB - A new Dactylella species, Dactylella alba was isolated from the ascospores of Orbilia alba collected in Wenshan County, Yunnan Province, China. Conidiophores were either not branched or occasionally branched, bearing divergent sterigmata on the tip with single conidium on each. Conidia were elongated ellipsoids, 1-2 septate, mostly 1 septate. By combining the ITS sequence with morphological characteristics, a new anamorphic species is described and illustrated together with its teleomorph. PMID- 19557344 TI - Effect of glycosylation on the biochemical properties of beta-xylosidases from Aspergillus versicolor. AB - Aspergillus versicolor grown on xylan or xylose produces two beta-xylosidases with differences in biochemical properties and degree of glycosylation. We investigated the alterations in the biochemical properties of these beta xylosidases after deglycosylation with Endo-H or PNGase F. After deglycosylation, both enzymes migrated faster in PAGE or SDS-PAGE exhibiting the same R(f). Temperature optimum of xylan-induced and xylose-induced beta-xylosidases was 45 degrees C and 40 degrees C, respectively, and 35 degrees C after deglycosylation. The xylan-induced enzyme was more active at acidic pH. After deglycosylation, both enzymes had the same pH optimum of 6.0. Thermal resistance at 55 degrees C showed half-life of 15 min and 9 min for xylose- and xylan-induced enzymes, respectively. After deglycosylation, both enzymes exhibited half-lives of 7.5 min. Native enzymes exhibited different responses to ions, while deglycosylated enzymes exhibited identical responses. Limited proteolysis yielded similar polypeptide profiles for the deglycosylated enzymes, suggesting a common polypeptide core with differential glycosylation apparently responsible for their biochemical and biophysical differences. PMID- 19557345 TI - Molecular analysis of the copper-responsive CopRSCD of a pathogenic Pseudomonas fluorescens strain. AB - CopRS/CopABCD is one of the known systems that control copper homeostasis in bacteria. Although CopRS/CopABCD homologues are found to exist in Pseudomonas fluorescens, the potential role of this system in P. fluorescens has not been investigated. In this study a genetic cluster, consisting of copR, S, C, and D but lacking copAB, was identified in a pathogenic P. fluorescens strain (TSS) isolated from diseased fish. The copRSCD cluster was demonstrated to be required for full copper resistance and regulated at the transcription level by Cu. Expression of copCD is regulated directly by the two-component response regulator CopR, which also regulates its own expression. Interruption of the regulated expression of copR affected bacterial growth, biofilm formation, and tissue dissemination and survival. A mutant CopR, which lacks the N-terminal signal receiver domain and is constitutively active, was found to have an attenuating effect on bacterial virulence when expressed in TSS. To our knowledge, this is the first report that suggests a link between CopR and bacterial pathogenicity in P. fluorescens. PMID- 19557346 TI - Bradyrhizobium spp. and Sinorhizobium fredii are predominant in root nodules of Vigna angularis, a native legume crop in the subtropical region of China. AB - Adzuki bean (Vigna angularis) is an important legume crop native to China, but its rhizobia have not been well characterized. In the present study, a total of 60 rhizobial strains isolated from eight provinces of China were analyzed with amplified 16S rRNA gene RFLP, IGS-RFLP, and sequencing analyses of 16S rRNA, atpD, recA, and nodC genes. These strains were identified as genomic species within Rhizobium, Sinorhizobium, Mesorhizobium, Bradyrhizobium, and Ochrobactrum. The most abundant groups were Bradyrhizobium species and Sinorhizobium fredii. Diverse nodC genes were found in these strains, which were mainly co-evolved with the housekeeping genes, but a possible lateral transfer of nodC from Sinorhizobium to Rhizobium was found. Analyses of the genomic and symbiotic gene backgrounds showed that adzuki bean shared the same rhizobial gene pool with soybean (legume native to China) and the exotic Vigna species. All of these data demonstrated that nodule formation is the interaction of rhizobia, host plants, and environment characters. PMID- 19557348 TI - Gene expression analysis of Phanerochaete chrysosporium during the transition time from primary growth to secondary metabolism. AB - In order to identify the secondary metabolism-related genes of Phanerochaete chrysosporium growing under pure O2 and nitrogen-limited conditions, 2322 ESTs fragments originated from two suppression-subtractive libraries were analyzed using the cDNA microarray technique. Ten significantly upregulated and 22 significantly downregulated genes were identified in the 72 h cultured mycelia RNA samples (secondary metabolism). According to qPCR, 16 out of the 32 genes were expressed differently in secondary metabolism. Transcripts of secondary metabolism up-regulation genes exhibited homologies to aryl-alcohol dehydrogenase (SShl554), ABC transporter gene (SSH624), chitinase (SSH963), heat shock protein (SSH1193), catalase (SSH317), cytochrome P450 (SSH331), glucosamine-6-phosphate isomerase (SSH611), and alkyl hydroperoxide reductase (SSH362) genes. Ninety three genes could be classified by Eukaryotic Orthologous Groups (KOG). Among the genes assigned a function, gene expression patterns were different in both secondary metabolism and primary metabolism. In the group of "Cellular Processes and Signaling," most of the genes were from the primary metabolism library. On the other hand, genes from the secondary metabolism library were found mainly in the "Information Storage" and "Processing and Poorly Characterized" groups. Based on the KOG functional assignments, six genes belong to the ubiquitin system, and all of them were from primary metabolism phase. The presence of the H2O2-relevant genes suggested that parts of the genes expressed in 72 h might be involved in the ligninolytic process during secondary metabolism of P. chrysosporium. PMID- 19557347 TI - Expression and regulation of ribulose 1,5-bisphosphate carboxylase/oxygenase genes in Mycobacterium sp. strain JC1 DSM 3803. AB - Ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) is the key enzyme of the Calvin reductive pentose phosphate cycle. Two sets of structural genes (cbbLS 1 and -2) for form I RubisCO have been previously identified in the Mycobacterium sp. strain JC1, which is able to grow on carbon monoxide (CO) or methanol as sole sources of carbon and energy. Northern blot and reverse transcriptase PCR showed that the cbbLS-1 and -2 genes are expressed in cells grown on either carbon monoxide (CO) or methanol, but not in cells grown in nutrient broth. A promoter assay revealed that the cbbLS-2 promoter has a higher activity than the cbbLS-1 promoter in both CO- and methanol-grown cells, and that the activities of both promoters were higher in CO-grown cells than in methanol-grown cells. A gel mobility shift assay and footprinting assays showed that CbbR expressed in Escherichia coli from a cbbR gene, which is located downstream of cbbLS-1 and transcribed in the same orientation as that of the cbbLS genes, specifically bound to the promoter regions of the cbbLS-1 and -2 genes containing inverted repeat sequence. A DNase I footprinting assay revealed that CbbR protected positions -59 to -3 and -119 to -78 of the cbbLS-1 and -2 promoters, respectively. Overexpression of CbbR induced the transcription of RubisCO genes in Mycobacterium sp. strain JC1 grown in nutrient broth. Our results suggest that the CbbR product from a single cbbR gene may positively regulate two cbbLS operons in the Mycobacterium sp. strain JC1 as is the case for Rhodobacter sphaeroides and Cupriavidus necator. PMID- 19557349 TI - The gene bap, involved in biofilm production, is present in Staphylococcus spp. strains from nosocomial infections. AB - This study analyzed ten strains of coagulase-negative staphylococci (CNS) involved in nosocomial infections in three Brazilian hospitals. Their antibiotic susceptibility profile showed that most strains exhibited multiple antibiotic resistance and possessed the mecA gene. The ability of these strains to adhere to polystyrene microtiter plates was also tested and nine of them proved to be biofilm producers at least in one of the three conditions tested: growth in TSB, in TSB supplemented with NaCl, or in TSB supplemented with glucose. The presence of the bap gene, which codes for the biofilm-associated protein (Bap), was investigated in all ten strains by PCR. AU strains were bop-positive and DNA sequencing experiments confirmed that the fragments amplified were indeed part of a bap gene. The presence of the icaA gene, one of the genes involved in polysaccharide intercellular adhesin (PIA) formation, was also detected by PCR in eight of the ten strains tested. The two icaA-negative strains were either weak biofilm producer or no biofilm producer, although they were bop-positive. To our knowledge, this is the first report demonstrating the presence of the bap gene in nosocomial isolates of CNS, being also the first report on the presence of this gene in Staphylococcus haemolyticus and S. cohnii. PMID- 19557350 TI - Mobilization functions of the bacteriocinogenic plasmid pRJ6 of Staphylococcus aureus. AB - Plasmid pRJ6 is the first known bacteriocinogenic mobilizable (Mob) plasmid of Staphylococcus aureus. Its Mob region is composed of four mob genes (mobCDAB) arranged as an operon, a genetic organization uncommon among S. aureus Mob plasmids. oriT (pRJ6) was detected in a region of 431 bp, positioned immediately upstream of mobC. This region, when cloned into pCN37, was able to confer mobilization to the re-combinant plasmid only in the presence of pRJ6. The entire Mob region, including oriT (pRJ6), is much more similar to Mob regions from several coagulase-negative staphylococci plasmids, although some remarkable similarities with S. aureus Mob plasmids can also be noted. These similarities include the presence within oriT (pRJ6) of the three mcb (MobC binding sites), firstly described in pC221 and pC223, an identical nick site also found in these same plasmids, and a nearly identical sra(pC223) site (sequence recognized by MobA). pRJ6 was successfully transferred to S. epidermidis by conjugation in the presence of the conjugative plasmid pGOl. Altogether these findings suggest that pRJ6 might have been originally a coagulase-negative staphylococci plasmid that had been transferred successfully to S. aureus. PMID- 19557351 TI - Nup211, the fission yeast homolog of Mlp1/Tpr, is involved in mRNA export. AB - Synthetic lethal mutants have been previously isolated in fission yeast Schizosaccharomyces pombe, which genetically interact with spmex67, in order to identify the genes involved in mRNA export. The nup211 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMex2, under synthetic lethal condition. We showed that Nup211, fission yeast homolog of Mlpl/Mlp2/Tpr, is essential for vegetative growth and Nup211-GFP proteins expressed at endogenous level are localized mainly in nuclear periphery. The accumulation of poly(A)(+) RNA in the nucleus is exhibited when expression of nup211 is repressed or over-expressed. These results suggest that the Nup211 protein plays a pivotal role of mRNA export in fission yeast. PMID- 19557352 TI - Exchange of the VP5 of infectious bursal disease virus in a serotype I strain with that of a serotype II strain reduced the viral replication and cytotoxicity. AB - Infectious bursal disease virus (IBDV), belonging to Avibirnavirus genus in the Birnaviridae family, consists of two segments of double-strand RNA. There are two distinct serotypes of IBDV, the pathogenic serotype I and the non-pathogenic serotype II. Comparison of the deduced amino acid sequences of a panel of VP5 genes retrieved from GenBank revealed a high identity among strains within the serotype I or serotype II group but a low identity between strains across two serotypes. In this study, we rescued two mosaic viruses, rGtGxVP5 and rGt2382VP5 by exchanging the VP5 gene of a cell culture-adapted serotype I Gt strain with its counterpart of the very virulent IBDV Gx strain, or a non-pathogenic 23/82 strain of the serotype II. In comparison to the parental strain rGt virus, the rGtGxVP5 showed the similar viral replication, cytotoxicity and the ability of inducing apoptosis; however, the other mosaic virus rGt2382VP5 had a lower titer and a reduced cytotoxicity. Although exchange of VP5 within serotype I group did not alter the viral replication and cytotoxicity of Gt strain, exchange of VP5 in the serotype I with that of a serotype II reduced the viral replication and cytotoxicity on chicken embryo fibroblast (CEF) cells. Therefore, the VP5 of serotype II may be one of the factors responsible for the distinct pathogenic features of two serotypes. PMID- 19557353 TI - Cloning, expression, and characterization of xylose reductase with higher activity from Candida tropicalis. AB - Xylose reductase (XR) is a key enzyme in xylose metabolism because it catalyzes the reduction of xylose to xylitol. In order to study the characteristics of XR from Candida tropicalis SCTCC 300249, its XR gene (xyll) was cloned and expressed in Escherichia coli BL21 (DE3). The fusion protein was purified effectively by Ni2+-chelating chromatography, and the kinetics of the recombinant XR was investigated. The Km values of the C. tropicalis XR for NADPH and NADH were 45.5 microM and 161.9 microM, respectively, which demonstrated that this XR had dual coenzyme specificity. Moreover, this XR showed the highest catalytic efficiency (kcat =1.44 x 10(4) min(-1)) for xylose among the characterized aldose reductases. Batch fermentation was performed with Saccharomyces serivisiae W303 lA:pYES2XR, and resulted in 7.63 g/L cell mass, 93.67 g/L xylitol, and 2.34 g/L x h xylitol productivity. This XR coupled with its dual coenzyme specificity, high activity, and catalytic efficiency proved its utility in in vitro xylitol production. PMID- 19557354 TI - Antibacterial activity of recombinant hCAP18/LL37 protein secreted from Pichia pastoris. AB - Human antimicrobial peptide CAP18/LL37 (hCAP18/LL37) was expressed in Pichia pastoris and its antibacterial activity was tested against pathogenic bacteria. The full length ORF of hCAP18/LL37 was cloned into the pPICZaA vector followed by integration into the genomic AOX1 gene of P. pastoris. Agar diffusion assay demonstrated that the different hCAP18/LL37 transformants showed various antibacterial activities against Staphylococcus aureus, Micrococcus luteus, and Salmonella gastroenteritis. The secreted form of hCAP18/LL37 exhibited its maximum activity after 72 h incubation with 2% methanol in MM media, not in BMM. This result suggests that the yeast secreted expression system can be used as a production tool of antimicrobial peptides for industrial or pharmaceutical application. PMID- 19557356 TI - Synthesis and biological evaluation of 2,5-diaminobenzamide derivatives as anti proliferating agents. AB - The screening of the chemical library for the anti-proliferative activity of the chemical library provided 2,5-diaminobenzamide as the initial hit. The confirmation and the optimization of hit were performed by synthesis followed by the evaluation of growth inhibitory activity against human cancer cell lines. The most active growth inhibitor showed IC(50) of 1.0 microM. The compound 7 increased not only sub-G1 population but also number of cells which are stained with Annexin V-FITC and 7-AAD, suggesting that compound 7 induced cell death is apoptosis. PMID- 19557357 TI - Inhibitory effect of saponin fraction from Codonopsis lanceolata on immune cell mediated inflammatory responses. AB - Saponin components are known to be pharmaceutically, cosmetically and nutraceutically valuable principles found in various herbal medicine. In this study, we evaluated the inhibitory role of saponin fraction (SF), prepared from C. lanceolata, an ethnopharmacologically famous plant, on various inflammatory responses managed by monocytes, macrophages, lymphocytes and mast cells. SF clearly suppressed the release of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha, but not prostaglandin E(2) (PGE(2)). While this fraction did not scavenge the reactivity of SNP-induced radicals in RAW264. 7 cells, it negatively modulated the phagocytic uptake of macrophages treated with FITC-dextran. Interestingly, SF completely diminished cell-cell adhesion events induced by both CD29 and CD43, but not cell-fibronectin adhesion. Concanavalin (Con) A [as well phytohemaglutinin A (PHA)]-induced proliferation of splenic lymphocytes as well as interferon (IFN)-gamma production were also clearly suppressed by SF treatment. Finally, SF also significantly blocked the degranulation process of mast cell line RBL-2H3 cell as assessed by DNP-BSA-induced beta-hexosaminidase activity. The anti-inflammatory activities of SF on NO production seemed to be due to inhibition of nuclear factor (NF)-kappaB activation signaling, since it blocked the phosphorylation of inhibitor of kappaB (IkappaB)alpha as well as inducible NO synthase (iNOS) expression. Therefore, these results suggest that SF may be considered as a promising herbal medicine with potent anti-inflammatory actions. PMID- 19557358 TI - Anti-allergic effects of white rose petal extract and anti-atopic properties of its hexane fraction. AB - Rosa rugosa is a species of rose native to eastern Asia. The root of R. rugosa has been used to treat diabetes mellitus, pain and chronic inflammatory disease, and a R. rugosa petal extract has a strong anti-oxidant effect. In the present study, we examined if solvent fractions from white rose petal extract (WRPE) had any anti-allergic or anti-atopic effects not previously reported. WRPE and butanol and hexane fractions effectively reduced systemic anaphylactic reactions and anti-dinitrophenyl (DNP) IgE-mediated passive cutaneous anaphylaxis in mice, with the greatest inhibition observed for the hexane fraction. In addition, a significant reduction of scratching behavior by mice after histamine injection suggested this fraction's potential anti-allergic effect. At the cell level, the hexane fraction markedly inhibited beta-hexosaminidase release from RBL-2H3 mast cells and suppressed the expressions of mRNA interferon-gamma and interleukin-4 cytokines produced by T helper cells (type 1 and 2). These results strongly support that the hexane fraction may have an effect on atopic dermatitis, as these 2 cell types play central roles in the pathogenesis of atopic dermatitis. In conclusion, these results suggest that either the hexane fraction or one of its components may be beneficial for the treatment of allergic diseases, including atopic dermatitis. PMID- 19557359 TI - Isolation and tandem mass fragmentations of an anti-inflammatory compound from Aralia elata. AB - One-step isolation of a saponin from Aralia elata was undertaken using high-speed countercurrent chromatography coupled with evaporative light scattering detection. A triterpenoid saponin, elatoside F, was purified with 96.8% purity using a two-phase-system comprising chloroform-methanol-water-isopropanol. The yield was 35.0 mg from 348.2 mg of the enriched saponin fraction. In vitro anti inflammatory study demonstrated that elatoside F inhibited lipopolysaccharide induced nitric oxide production, as well as nuclear factor kappaB activation, in a dose-dependent manner. Two types of mass ionization technique were compared on elatoside F to investigate characteristic fragmentation patterns. MALDI-TOF tandem mass spectrometric fragmentation patterns of sodiated ions provided structural information on glycosidic cleavages and on extensive cross-ring cleavages. Electrospray ionization multiple-stage tandem mass fragmentation of both sodiated and lithiated ions could provide information on glycosidic cleavages. All observed tandem mass fragmentation spectra provided valuable elatoside F structural information when unknown samples from crude extracts are under screening by mass spectrometry. PMID- 19557360 TI - A new cytotoxic spiroketal from the myxobacterium Sorangium cellulosum. AB - A new spiroketal, spirodienal B (1), was isolated from culture extracts of S. cellulosum. Its structure was elucidated by spectroscopic analysis. In addition to 1, the strain produced a second, closely related antibiotic, spirodienal. Both compounds demonstrated potent cytotoxicity against human tumor cells. Against human tumor cells such as HCT-15, the activity of 1 was more than 4000 times stronger than that of doxorubicin in terms of IC(50). PMID- 19557361 TI - Bioactive triterpenoids from Callistemon lanceolatus. AB - A new triterpenoid, 30-hydroxyalphitolic acid 1, and eight known triterpenoids, alphitolic acid 2, lupenol 3, 3-acetoxy-olean-18-en-28-oic acid 4, betulinic acid 5, ursolic acid 6, betulinic acid 3-O-caffeate 7, morolic acid 3-O-caffeate 8, and ursolic acid 3-O-caffeate 9, were isolated from Callistemon lanceolatus. Their structures were determined using spectroscopic techniques, which included 1D- and 2D-NMR. All compounds were evaluated for the inhibition of LPS-induced nitric oxide production in murine macrophage RAW264.7 cells. Betulinic acid 3-O caffeate 7 showed a moderate inhibitory effect on nitric oxide production with IC(50) value of 15.4 microM. PMID- 19557362 TI - Four flavonoids from Echinosophora koreensis and their effects on alcohol metabolizing enzymes. AB - A new prenylated dihydroflavonol, 3-hydroxy-kenusanone B 1, as well as three other known isoflavanones, sophoronol 2, sophoraisoflavanone A 3 and kenusanone H 4, were isolated from the rhizomes of Echinosophora koreensis. The structures of these compounds were elucidated using spectroscopic analyses that included extensive 2D NMR, optical rotation spectrometry and mass spectrometry. All four flavonoids enhanced the activities of alcohol metabolizing enzymes such as alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) at micromolar concentrations. Sophoronol 2 showed a nine-fold increased activation of alcohol dehydrogenase and aldehyde dehydrogenase than a negative control group at concentrations of 100 microg/mL and 50 microg/mL, respectively. This study suggests that prenylated flavonoids have the potential to prevent 'hangovers' after alcohol intake. PMID- 19557363 TI - Bile acid derivatives from a sponge-associated bacterium Psychrobacter sp. AB - In our search for bioactive metabolites from a marine sponge-associated bacterium Psychrobacter sp., a new bile acid derivative (1), which was assumed to be an artifact, were isolated along with six known (2-7) compounds by bioactivity guided fractionation. Elucidation of the structure of the new compound was done using a combination of NMR ((1)H, (13)C, HMBC, HSQC, and COSY) and MS spectroscopy. Compound 1 exhibited moderate suppressive effects on both NO and IL 6 production at a concentration of 200 microM (87.3 microg/mL) without significant cytotoxicity against cells. Compounds 2-5 and 7 showed selective inhibitory activity against several human pathogenic bacterial strains at the low concentration of 30 microg/well. In a cytotoxicity evaluation, only compound 7 showed mild cytotoxicity against five human solid tumor cell lines (A-549, SK-OV 3, SK-MEL-2, XF-498, and HCT-15) with ED(50) values in the range of 11-14 microg/mL. PMID- 19557364 TI - Two new secoiridoid glycosides from the rhizomes of Gentiana scabra Bunge. AB - Two new secoiridoid glycosides, 4'''-O-beta-D-glucopyranosyltrifloroside 1 and 4'''-O-beta-D-glucopyranosylscabraside 2, along with three known secoiridoids were isolated from the rhizomes of Gentiana scabra (Gentianaceae) in our recent phytochemical study. Their chemical structures were determined by spectroscopic data including 1D and 2D NMR spectra. The chemotaxonomic significance of the secoiridoid glycosides is briefly discussed. PMID- 19557365 TI - Neuroprotective effect of epigallocatechin-3-gallate against beta-amyloid-induced oxidative and nitrosative cell death via augmentation of antioxidant defense capacity. AB - beta-Amyloid (Abeta) peptide, a major component of senile plaques has been regarded to play a crucial role in the development and neuropathogenesis of Alzheimer's disease (AD). Increasing data from in vitro and in vivo studies indicate that Abeta-induced damages in neurons and glia are mediated via nitrosative as well as oxidative stress. Therefore, recent researches have been focused on searching for dietary and herbal manipulations to protect against the Abeta-induced oxidative and/or nitrosative cell death. Epigallocatechin-3-gallate (EGCG), one of these candidates is a major polyphenolic compound present in green tea and has been reported to exhibit potent antioxidant and anti-inflammatory properties. In the present study, we have investigated the effect of EGCG against Abeta-induced oxidative and/or nitrosative cell death in BV2 microglia. Abeta treatment led to apoptosis in BV2 cells as revealed by DNA fragmentation, perturbation of mitochondrial transmembrane potential, and alterations in the expression of apoptosis-regulator Bcl-2 family proteins. EGCG pretreatment effectively ameliorated Abeta-induced cytotoxicity and manifestation of proapoptotic signals. Furthermore, BV2 cells exposed to Abeta underwent nitrosative stress as shown by the increased expression of inducible nitric oxide synthase (iNOS) and subsequent production of nitric oxide (NO) and peroxynitrite, which were effectively suppressed by EGCG pretreatment. To elucidate a molecular mechanism underlying the neuroprotective effect of EGCG, we have examined the cellular metabolism of reduced glutathione (GSH) with antioxidant properties. EGCG treatment fortified cellular GSH pool through elevated mRNA expression of gamma-glutamylcysteine ligase (GCL), the rate limiting enzyme in the glutathione biosynthesis. These results suggest that EGCG may have preventive and/or therapeutic potential in AD patients by augmenting cellular antioxidant defense capacity and attenuating Abeta-mediated oxidative and/or nitrosative cell death. PMID- 19557366 TI - Cadmium adaptation is regulated by multidrug resistance-associated protein mediated Akt pathway and metallothionein induction. AB - To elucidate the mechanisms involved in adaptation of lung epithelial cells to cadmium (Cd), we established a cell line that exhibits Cd-resistance (RWI38). RWI38 showed approximately 5-fold greater Cd-resistance (MTT assays) than WI38 cells, and cross-resistance to Zn and cisplatin. RWI38 cells also demonstrated an upregulated level of multidrug resistance-associated protein (MRP) and metallothionein (MT) (as shown by Western blot analysis and RT-PCR studies). The protein level of MRP decreased after Cd exposure in WI38 cells, but was sustained at high levels in RWI38 cells, leading led to enhanced calcein efflux. Cd induced Akt phosphorylation in RWI38 but not WI38 cells; this was prevented by probenecid or siRNA for MRP, both of which led to enhanced cell death, as demonstrated by capsase-3 activation and decreased cell viability. These results suggest a functional role for MRP in the regulation of the Akt pathway as well in the efflux pumping of drugs, thereby contributing toward the adaptation of cells to Cd toxicity. The findings of this study could be potentially beneficial in the design of therapeutic targets for Cd-induced tumor progression. PMID- 19557367 TI - Physicochemical characterization of tacrolimus-loaded solid dispersion with sodium carboxylmethyl cellulose and sodium lauryl sulfate. AB - To develop a novel tacrolimus-loaded solid dispersion with improved solubility, various solid dispersions were prepared with various ratios of water, sodium lauryl sulfate, citric acid and carboxylmethylcellulose-Na using spray drying technique. The physicochemical properties of solid dispersions were investigated using scanning electron microscopy, differential scanning calorimetery and powder X-ray diffraction. Furthermore, their solubility and dissolution were evaluated compared to drug powder. The solid dispersion at the tacrolimus/CMC-Na/sodium lauryl sulfate/citric acid ratio of 3/24/3/0.2 significantly improved the drug solubility and dissolution compared to powder. The scanning electron microscopy result suggested that carriers might be attached to the surface of drug in this solid dispersion. Unlike traditional solid dispersion systems, the crystal form of drug in this solid dispersion could not be converted to amorphous form, which was confirmed by the analysis of DSC and powder X-ray diffraction. Thus, the solid dispersion system with water, sodium lauryl sulfate, citric acid and CMC-Na should be a potential candidate for delivering a poorly water-soluble tacrolimus with enhanced solubility and no convertible crystalline. PMID- 19557368 TI - Antimalarial effect of N-acetyl-L-Leucyl-L-leucyl-L-norleucinal by the inhibition of Plasmodium falciparum Calpain. AB - The biological understanding of malaria parasites has increased considerably over the past two decades with the discovery of many potential targets for the development of new antimalarial drugs. Calpain, a cysteine protease of Plasmodium falciparum, is believed to be a central mediator essential for parasitic activity. However, the utility of calpain as a potential anti-malarial target in P. falciparum has not been fully determined. In the present study, we determined the effect of N-acetyl-L-Leucyl-L-leucyl-L-norleucinal (ALLN)-treatment on the expression of calpain in erythrocytic stages of P. falciparum and its usefulness as an antimalarial chemotherapeutic agent. ALLN was shown to have low toxicity to HeLa cells but high toxicity to malaria. ALLN inhibited the expression of calpain in ring, trophozoite and schizont stages when treated for 48 h. Also, after 48 h, samples were characterized by 6.15% and 0% parasitemia without ALLN treatment and with ALLN treatment, respectively. Brightfield and confocal microscopy revealed that ALLN treatment affects merozoite maturation. As ALLN concentration increased from 1 muM to 100 microM, ring stage parasites did not mature into the schizont stage. When ALLN treatment was continued for 48 h, it also significantly inhibited the maturation of ring-stage parasites into trophozoite or schizont stages and survival of malarial parasites. Taken together, these findings suggest that ALLN inhibit the maturation and survival of P. falciparum and calpain expression, and thus has potential utility as an antimalarial chemotherapeutic agent. PMID- 19557369 TI - Effect of coenzyme Q10 on cutaneous healing in skin-incised mice. AB - Coenzyme Q10 (CoQ10) is a biosynthesized quinone with 10 isoprene side chains in humans. To investigate the anti-inflammatory and wound healing effect of CoQ10, we performed in vivo and in vitro experiments. In vivo studies, there were 3 groups; Naive (without skin incision), Control (with skin incision) and CoQ10 (100 mg/kg treatment with skin incision). Collagen-like polymer (CLP) level of CoQ10 group was increased significantly compared to the control group (p<0.05). Also, CoQ10 group showed significant inhibition on myeloperoxidase (MPO) and PLA(2) level compared to the control group (p<0.05). These data show that CoQ10 may have an anti-inflammatory and a wound healing effect. CoQ10 showed significant antioxidant activity in vivo on malondialdehyde (MDA) and superoxide dismutase (SOD) levels compared to the control group (p<0.05). Although CoQ10 did not show antioxidant activity in cell free system of DPPH radical scavenge, it had a potent antioxidant activity in cell culture system of both silica- and zymosan-induced reactive oxygen species generation using Raw 264.7 cells. This result may be associated with the conversion of CoQ10 to the reduced form (CoQ10H(2)) in the presence of some kinds of intracellular reducing agents. In conclusion, it is considered that CoQ10 appears to have a cutaneous healing effect in vivo, which may be related to the secondary action of CoQ10. PMID- 19557370 TI - Anti-tumor activity of noble indirubin derivatives in human solid tumor models in vitro. AB - Indirubin has been identified as a component of a traditional Chinese medicine, Danggui Longhui Wan, which is used for the treatment of chronic myelogenous leukemia. Indirubin inhibits cyclin-dependent kinases (CDKs) and induces cell cycle arrest and apoptosis in cancer cells. Many indirubin derivatives have been studied for their potential anti-solid tumor activity. We have synthesized and evaluated many indirubin derivatives. In order to compare and confirm the potential of our major derivatives as anti-solid tumor agents, we examined their anti-proliferative activity in monolayers, as well as in multicellular spheroids (MCS) cultures of human colorectal cancer cells, DLD-1 and HT-29. The MCS model is an in vitro solid tumor model that is increasingly used for the evaluation of anti-solid tumor activity. 5-nitro-indirubin-3'-oxime (4c) and 5'-bromo-5-nitro indirubin-3'-oxime (4l), compared to 5-trimethylacetamido-indirubin-3'-oxime (11) and 5-diphenylacetamido-indirubin-3'-oxime (33) showed greater anti-proliferative effects in monolayers, but lower anti-proliferative effects in MCS. Overall, our data suggest that compounds 11 and 33 may exert a significant anti-solid tumor activity via a mechanism other than CDK inhibition, different from that of 4c and 4l. These compounds are worth further investigation with respect to their anti solid tumor activity and their mechanism of action in various solid tumor models. PMID- 19557371 TI - Anti-ischemic activities of aralia cordata and its active component, oleanolic acid. AB - Aralia has been reported to exhibit various pharmacological properties, including anti-inflammatory, antidiabetic and antioxidant activities. We performed in vitro and in vivo analyses on the neuroprotective effects of an ethanolic extract of the aerial parts of Aralia cordata Thunb. (Araliaceae). In cultured cortical neurons from rats, A. cordata (5-20 microg/mL) inhibited 100 muM hydrogen peroxide (H(2)O(2))-induced apoptotic neuronal death, elevation of intracellular calcium concentration ([Ca(2+)](i)) and generation of reactive oxygen species (ROS). Since oleanolic acid isolated from A. cordata also inhibited H(2)O(2) induced neuronal death, increase in [Ca(2+)](i) and ROS generation in cultured cortical neurons, some of the neuroprotective effects of A. cordata might be attributable to this compound. In rats, A. cordata prevented cerebral ischemic injury induced by 3 h of middle cerebral artery occlusion, followed by 24 h of reperfusion. Ischemic infarct and edema volumes were significantly reduced in rats that received A. cordata (50 mg/kg, orally). These animals exhibited a corresponding improvement in neurological function and a reduction of neuronal death, as determined histologically from the cortex and hippocampal regions. It is possible that the anti-oxidative properties of A. cordata may be responsible for its neuroprotective effects against focal cerebral ischemic injury. In future, A. cordata might play a therapeutic role in the prevention and treatment of neurodegeneration in stroke. PMID- 19557372 TI - Crystal forms of atorvastatin. AB - The objective of this work was to investigate the existence of new polymorphs and pseudopolymorphs of atorvastatin and the transformation of crystal forms. Three crystal forms of atorvastatin have been isolated by recrystallization and characterized by powder X-ray diffractometry (PXRD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TG). The PXRD and DSC patterns of the three crystal forms were different respectively and it was confirmed that Form 3 is a new crystal form. After storage of 2 years at room temperature, this new crystal form was not transformed and it was shown to have a good physical stability. PMID- 19557373 TI - The analgesic effect of decursinol. AB - Although decursinol, which is one of the coumarins purified from the dried roots of Angelica gigas Nakai, was previously demonstrated to have antinociceptive effects on various mouse pain models such as tail-flick, hot-plate, formalin, writhing, and several cytokine-induced pain tests, the possible involvement of its analgesic effects and non-steroidal anti-inflammatory drugs (NSAIDs) has not been clearly elucidated yet. In this study, we characterized the possible interaction between decursinol and aspirin or acetaminophen in the writhing test. The antinociceptive effects of decursinol were observed at an orally-administered dose of 50 mg/kg but not at 25 or 10 mg/kg. In addition, the analgesic effects of aspirin (ASA) and acetaminophen (APAP) were shown at an orally-administered dose of 200 mg/kg but not at 50 or 100 mg/kg. We examined the effects of decursinol on the ASA or APAP at sub-analgesic doses. Although the co-administration of decursinol and ASA did not show any differences at doses of 10 or 25 mg/kg and 50 or 100 mg/kg, respectively, synergistic effects between decursinol and APAP were observed in the group of decursinol (25 mg/kg) and APAP (100 mg/kg) co administration. These results indicated that the analgesic effect of decursinol might be involved in supraspinal cyclooxygenase regulation that might be overlapped with APAP-induced analgesic mechanisms rather than systemic or peripheral prostaglandin modulation. PMID- 19557374 TI - Inhibitory effects of oligopeptides from hen egg white on both human platelet aggregation and blood coagulation. AB - Egg white proteins have many biological functions and substantial nutritional benefits when used as a food source; however, they also contain allergens such as ovalbumin, ovomucoid, and ovotransferrin. We prepared oligopeptides without allergens from hen egg whites via the use of several proteases, and assessed their effects on platelet aggregation and blood coagulation, known to both of which are known to be major risk factors in thrombogenesis. Egg white oligopeptides (EWOP) inhibited collagen-induced human platelet aggregation in a dose-dependent manner. Additionally, we attempted to determine whether cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), aggregation-inhibiting intracellular molecules, regulate EWOP-inhibited platelet aggregation. EWOP caused an increase in cAMP levels, but did not affect cGMP levels, which suggests that the anti-platelet activity of EWOP operates in a cAMP dependent manner, rather than via a cGMP-dependent process, in collagen-induced platelet aggregation. In addition, EWOP induced a significantly prolonged prothrombin time (PT) as compared with the controls. These data show that EWOP inhibits the conversion of fibrinogen to fibrin in a plasmatic atmosphere on an extrinsic pathway. Accordingly, these findings suggest that EWOP may be an excellent candidate as a crucial inhibitor of platelet activation, and its anti platelet effects appear to involve the inhibition of both platelet aggregation and blood coagulation within the cardiovascular system. PMID- 19557375 TI - Synergistic decrease in blood pressure by captopril combined with losartan in spontaneous hypertensive rats. AB - This study was designed to examine the anti-hypertensive effect of the combination therapy of captopril with losartan by oral administration using both independent and cross-over experimental protocols. In independent experimental protocols, four different groups of spontaneous hypertensive rats (SHR) were treated for 1 or 2 weeks: control, captopril (20 mg/kg/day), losartan (20 mg/kg/day), and combination captopril (10 mg/kg/day) with losartan (10 mg/kg/day). In cross-over protocols, each SHR received all four treatments for 1 or 3 days with an interval of several days between each injection for washing-out and return to high blood pressure (BP) levels. BP and heart rate (HR) were measured in conscious telemetered SHR. According to the results from the independent protocol, regardless of a 1- or 2-week administration period, combination therapy with low doses of captopril and losartan had a greater anti hypertensive effect than individual high-dose monotherapy. Similarly, results from the cross-over protocol showed that regardless of 1-day or 3-day administration, the decrease in BP in the 11(th) and 12(th) hour after administration was greatest with the combination of low-dose captopril and losartan. Therefore, combination therapy with low doses of captopril with losartan lowered BP to a greater extent than a high dose of either individual monotherapy. PMID- 19557376 TI - Correction to "The retention behavior of ginsenosides in HPLC and its application to quality assessment of radix ginseng". PMID- 19557378 TI - [Intoxication with cyclopentolate eye drops]. AB - We report on an accidental intoxication with cyclopentolate eye drops. A 90-year old patient became confused and was admitted to the emergency department. His symptoms consisted of disorientation, ataxia, and psychomotor agitation. Similar cases have been described in the literature. With this case report we would like to draw attention to this little known differential diagnosis when confronted with confused patients. PMID- 19557377 TI - [Open reduction and dorsal spondylodesis C0-C2 in atlanto-occipital dislocation]. AB - We report the case of a 73-year-old male patient who was suffered trauma after a syncopal fall onto a railway track in the form of an atlanto-occipital dislocation. The diagnostic revealed a bilateral fracture of the occipital condyles coupled with a ventral atlanto-occipital dislocation (Jeanneret type 4) and also an odontoid fracture (Anderson type 2). The patient underwent dorsal spondylodesis of C0-C2 in combination with Magerl's C1-C2 screw fixation. Pre operatively and postoperatively no neurological abnormalities were found. This rarely occurring and survived traumatological situation is described using the present case as an example. PMID- 19557379 TI - [Emotional and affective disorders in epilepsy]. AB - The wide spectrum of comorbid mental disorders in epilepsy includes anxiety, affective, and personality disorders and psychosis. While the prevalence of mental disorders in the general epileptic population is listed at 6%, this rate is considerably higher in focal epilepsies, especially temporal lobe epilepsy, and the numbers given in the literature range from 20% to 70%, of which anxiety and depression are the most prominent. According to the Diathesis Stress Model, the effects of previously existing vulnerability and neurobiologic, iatrogenic, and psychosocial stress factors vary in the development of mental disorders in epilepsy. Roles are also played by learned reactions in responce to psychosocial stress as well as structural and functional disturbances in the limbic neuronal networks regulating affective, emotional, and social behaviors. Therapeutic measures may also contribute to the development of mental health problems. Several antiepileptic drugs have proven valuable in psychiatric treatment of mental disorders but also may have psychiatric side effects. Although established as a successful treatment option for focal epilepsies, surgery is also reported to influence mental health. PMID- 19557382 TI - Wheat-alien metaphase I pairing of individual wheat genomes and D genome chromosomes in interspecific hybrids between Triticum aestivum L. and Aegilops geniculata Roth. AB - Homoeologous metaphase I (MI) pairing of Triticum aestivum x Aegilops geniculata hybrids (2n = 5x = 35, ABDU(g)M(g)) has been examined by an in situ hybridization procedure permitting simultaneous discrimination of A, B, D and wild genomes. The seven D genome chromosomes (and their arms, except for 6D and 7D) plus some additional wheat chromosomes were also identified. Wheat-wild MI associations represented more than 60% of total, with an average ratio of 5:1:12 for those involving the A, B and D genomes, respectively. A remarkable between-chromosome variation for the level of wheat-wild genetic exchange is expected within each wheat genome. However, it can be concluded that 3DL and 5DL are the crop genome locations with the highest probability of being transferred to Ae. geniculata. Hybrids derived from the ph2b wheat mutant line showed increased MI pairing but identical pattern of homoeologous associations than those with active Ph2. PMID- 19557383 TI - Inheritance and molecular mapping of a downy mildew resistance gene, Pl (13) in cultivated sunflower (Helianthus annuus L.). AB - The inheritance of resistance to sunflower downy mildew (SDM) derived from HA-R5 conferring resistance to nine races of the pathogen has been determined and the new source has been designated as Pl ( 13 ) . The F(2) individuals and F(3) families of the cross HA-R5 (resistant) x HA 821 (susceptible) were screened against the four predominant SDM races 300, 700, 730, and 770 in separate tests which indicated dominant control by a single locus or a cluster of tightly linked genes. Bulked segregant analysis (BSA) was carried out on 116 F(2) individuals with 500 SSR primer pairs that resulted in the identification of 10 SSR markers of linkage groups 1 (9 markers) and 10 (1 marker) of the genetic map (Tang et al. in Theor Appl Genet 105:1124-1136, 2002) that distinguished the bulks. Of these, the SSR marker ORS 1008 of linkage group 10 was tightly linked (0.9 cM) to the Pl (13) gene. Genotyping the F(2) population and linkage analysis with 20 polymorphic primer pairs located on linkage group 10 failed to show linkage of the markers with downy mildew resistance and the ORS 1008 marker. Nevertheless, validation of polymorphic SSR markers of linkage group 1 along with six RFLP based STS markers of linkage group 12 of the RFLP map of Jan et al. (Theor Appl Genet 96:15-22, 1998) corresponding to linkage group 1 of the SSR map, mapped seven SSR markers (ORS 965-1, ORS 965-2, ORS 959, ORS 371, ORS 716, and ORS 605) including ORS 1008 and one STS marker (STS10D6) to linkage group 1 covering a genetic distance of 65.0 cM. The Pl (13) gene, as a different source with its location on linkage group 1, was flanked by ORS 1008 on one side at a distance of 0.9 cM and ORS 965-1 on another side at a distance of 5.8 cM. These closely linked markers to the Pl (13) gene provide a valuable basis for marker-assisted selection in sunflower breeding programs. PMID- 19557384 TI - IRS1 G972R polymorphism and type 2 diabetes: a paradigm for the difficult ascertainment of the contribution to disease susceptibility of 'low-frequency-low risk' variants. AB - AIMS/HYPOTHESIS: The aim of the study was to determine the association between IRS1 G972R polymorphism and type 2 diabetes; published data concerning this association have been conflicting. To obtain further insight into this topic, we performed a meta-analysis of all available case-control studies. METHODS: We performed a meta-analysis of 32 studies (12,076 cases and 11,285 controls). RESULTS: The relatively infrequent R972 variant was not significantly associated with type 2 diabetes (OR 1.09, 95% CI 0.96-1.23, p = 0.184 under a dominant model). Some evidence of heterogeneity was observed across studies (p = 0.1). In the 14 studies (9,713 individuals) in which the mean age at type 2 diabetes diagnosis was available, this variable explained 52% of the heterogeneity (p = 0.03). When these studies were subdivided into tertiles of mean age at diagnosis, the OR for diabetes was 1.48 (95% CI 1.17-1.87), 1.22 (95% CI 0.97-1.53) and 0.88 (95% CI 0.68-1.13) in the youngest, intermediate and oldest tertile, respectively (p = 0.0022 for trend of ORs). CONCLUSIONS/INTERPRETATION: Our findings illustrate the difficulties of ascertaining the contribution of 'low-frequency low-risk' variants to type 2 diabetes susceptibility. In the specific context of the R972 variant, approximately 200,000 study individuals would be needed to have 80% power to identify a 9% increase in diabetes risk at a genome-wide significance level. Under these circumstances, a strategy aimed at improving outcome definition and decreasing its heterogeneity may critically enhance our ability to detect genetic effects, thereby decreasing the required sample size. Our data suggest that focusing on early-onset diabetes, which is characterised by a stronger genetic background, may be part of such a strategy. PMID- 19557385 TI - Exercise intensity and insulin sensitivity: how low can you go? PMID- 19557386 TI - Glucose challenge test screening for prediabetes and undiagnosed diabetes. AB - AIMS/HYPOTHESIS: Diabetes prevention and care are limited by lack of screening. We hypothesised that screening could be done with a strategy similar to that used near-universally for gestational diabetes, i.e. a 50 g oral glucose challenge test (GCT) performed at any time of day, regardless of meal status, with one 1 h sample. METHODS: At a first visit, participants had random plasma and capillary glucose measured, followed by the GCT with plasma and capillary glucose (GCTplasma and GCTcap, respectively). At a second visit, participants had HbA(1c) measured and a diagnostic 75 g OGTT. RESULTS: The 1,573 participants had mean age of 48 years, BMI 30.3 kg/m(2) and 58% were women and 58% were black. Diabetes (defined by WHO) was present in 4.6% and prediabetes (defined as impaired glucose tolerance [2 h glucose 7.8-11.1 (140-199 mg/dl) with fasting glucose 45 in all ICU patients [OR, 0.54; 95% CI (0.39; 0.75); P = 0.0003]. Segmented regression analysis showed a 67.4% drop [95% CI (17.2%; 117.3%); P = 0.009] in the UTR rate after the first intervention was introduced. System factors played a major role in UTR occurrence. CONCLUSION: UTRs are common. A continuous quality-improvement program can reduce UTR rates in the ICU. PMID- 19557389 TI - Incidence and risk factors for ventilator-associated pneumonia after major heart surgery. AB - PURPOSE: Major heart surgery (MHS) patients are a particularly high-risk population for nosocomial infections. Our objective was to identify risk factors for ventilator-associated pneumonia (VAP) in patients undergoing MHS. METHODS: Prospective study including 1,844 patients operated from 2003 to 2006. RESULTS: Overall 106 patients (140 episodes) developed one or more episodes of VAP (5.7%, 22.2 episodes per 1,000 days of mechanical ventilation). VAP incidence was 45.9% in those patients requiring more than 48 h of MV. Enterobacteriaceae (32.8), Pseudomonas aeruginosa (28.6%) and Staphylococcus aureus (27.1%, of which 65.8% were methicillin resistant) were the principal microorganisms causing VAP. The independent risk factors for VAP were: age >70, perioperative transfusions, days of mechanical ventilation, reintubation, previous cardiac surgery, emergent surgery and intraoperative inotropic support. Median length of stay in the ICU for patients who developed VAP or not was, respectively, 25.5 versus 3 days (P < 0.001), and mortality was, respectively, 45.7 versus 2.8% in both populations (P < 0.001). We developed a predictive preoperative score with a sensitivity of 93% and a specificity of 40%. CONCLUSIONS: VAP is common in patients undergoing MHS that require more than 48 h of MV. In that "high-risk" population, innovative preventive measures should be developed and applied. PMID- 19557390 TI - Prevention of ventilator-associated pneumonia after cardiac surgery: prepare and defend! PMID- 19557391 TI - Double-bundle versus single-bundle ACL reconstruction using the horizontal femoral position: a prospective, randomized study. AB - The aim of this study was to evaluate whether anterior cruciate ligament (ACL) reconstruction using the double bundle technique (DB) improves stability in the knee compared with the single bundle technique (SB) with the femoral tunnel in a more horizontal position (2 or 10 o'clock). We conducted a randomized, prospective study. Forty patients were randomized to the DB group (20 patients) and the SB group (20 patients). Four-stranded semitendinosus and gracilis autologous grafts were used in the SB group and in the DB group the conventional four tunnel technique was carried out using the same tendons. The IKDC complete form was used for the preoperative evaluation, and in the follow-up the IKDC subjective knee evaluation form, IKDC current health assessment form and IKDC knee examination form were used. Anteroposterior (AP) laxity was evaluated by standardised and forced radiology in all patients. No significant preoperative between-group differences were found. During the follow-up, no differences were found between groups, except for significant between-group differences (P < 0.05) between the preoperative and postoperative evaluations. The IKDC index also showed significant differences in the 2-year follow-up. Median scores increased from 48 (range 41-54) to 81 (range 75-87) (P = 0.01) in the SB group and from 52 (range 46-58) to 80 (range 72-88) (P = 0.02) in the DB group. There were no significant differences between the groups in terms of functional scores. In conclusion, the 2 and 10 o'clock placements showed no significant differences between SB and DB techniques in the pivot-shift test, manual and radiological anterior posterior laxity and IKDC scores. However, significant between-group differences were found between the preoperative and postoperative evaluations. PMID- 19557392 TI - The relationship between bone marrow edema size and knee pain. AB - The purpose of our study was to determine the changes in the size of the edema observed on MRI scans and its relation to the activity pain of the patient and the rest pain in bone marrow edema (BME). A total of 51 patients were followed up at 3-month intervals for a period of 1 year. During the follow-ups, MRI scans of the patients' knees were obtained; the scores obtained on the Stanmore functional rating scale and visual analog scale were determined. The changes in these parameters and the correlation between them were examined. The following are the observations recorded during the bone marrow edema follow-ups: the size of the edema as observed on MRI scans decreased, and the activity pain and the rest pain decreased. While there is a correlation between the decrease in the edema size observed on MRI scans and decrease in the activity pain, there is no correlation between the decrease in the edema size observed on MRI scans and the decrease in rest pain. No changes were observed after a particular period of time with regard to decrease in the edema size observed on MRI scans, decrease in activity pain, and decrease in rest pain in follow-ups of BME patients. PMID- 19557394 TI - Anatomic double-bundle ACL reconstruction with femoral cortical bone bridge support using hamstrings. PMID- 19557395 TI - The reliability of a method for measuring the anterior cruciate ligament hamstring reflex: an objective assessment of functional knee instability. AB - Anterior cruciate ligament (ACL) rupture leads to mechanical and functional knee instability. Functional instability is likely attributable to a sensorimotor deficit. In previous studies, a method has been introduced which allows this deficit to be objectively assessed using ACL-hamstring reflex measurements. There is evidence that subjectively stable and unstable patients with ACL rupture can be objectively distinguished by this method. This study on 34 men was conducted to investigate the reliability of this method on the basis of several factors (intra-individual reproducibility, side-to-side differences, inter-examiner reliability, audiovisual stimuli, fatigue, height, weight, physical fitness, and level of activity at work). The role of gender was assessed in 20 men and 20 women. None of the factors had a relevant influence on reflex responses. The tests were reproducible and reliable. The method described here can thus be used for further research and the clinical diagnosis of functional instability following ACL rupture. PMID- 19557397 TI - Resonance ionization mass spectrometry of ion beam sputtered neutrals for element and isotope-selective analysis of plutonium in micro-particles. AB - Micro-particles containing actinides are of interest for risk assessments of contaminated areas, nuclear forensic analyses, and IAEA as well as Euratom safeguards programs. For their analysis, secondary ion mass spectrometry (SIMS) has been established as the state-of-the-art standard technique. In the case of actinide mixtures within the particles, however, SIMS suffers from isobaric interferences (e.g., (238)U/(238)Pu, (241)Am/(241)Pu). This can be eliminated by applying resonance ionization mass spectrometry which is based on stepwise resonant excitation and ionization of atoms with laser light, followed by mass spectrometric detection of the produced ions, combining high elemental selectivity with the analysis of isotopic compositions. This paper describes the instrumental modifications for coupling a commercial time-of-flight (TOF)-SIMS apparatus with three-step resonant post-ionization of the sputtered neutrals using a high-repetition-rate (kHz) Nd:YAG laser pumped tunable titanium:sapphire laser system. Spatially resolved ion images obtained from actinide-containing particles in TOF-SIMS mode demonstrate the capability for isotopic and spatial resolution. Results from three-step resonant post-ionization of bulk Gd and Pu samples successfully demonstrate the high elemental selectivity of this process. PMID- 19557398 TI - Imaging the cell wall of living single yeast cells using surface-enhanced Raman spectroscopy. AB - The surface of a living yeast cell (Saccharomyces cerevisiae strain W303-1A) has been labeled with silver (Ag) nanoparticles that can form nanoaggregates which have been shown to have surface-enhanced Raman scattering (SERS) activity. The cell wall of a single living yeast cell has been imaged by use of a Raman microspectroscope. The SERS spectra measured from different Ag nanoaggregates were found to be different. This can be explained on the basis of detailed spectral interpretation. The SERS spectral response originates from mannoproteins which cover the outermost regions of the yeast cell wall. Analysis of SERS spectra from the cell wall and the extracted mannoproteins from the yeast has been performed for the clarification of variation in SERS spectra. PMID- 19557396 TI - Antinociceptive effects of (O-methyl)-N-benzoyl tyramine (riparin I) from Aniba riparia (Nees) Mez (Lauraceae) in mice. AB - The present study examined the antinociceptive effects of (O-methyl) N-benzoyl tyramine (riparin I, ripI) isolated from the unripe fruit of Aniba riparia in chemical and thermal behavioral models of pain, such as acetic acid-induced abdominal writhing, formalin, and hot-plate tests in mice. Moreover, the involvement of the nitric oxide pathway as well as the opioid system in the antinociceptive action of ripI in the formalin test was investigated. RipI was administered both orally and intraperitoneally to male mice at single doses of 25 and 50 mg/kg. In the acetic acid-induced abdominal writhing, ripI decreased the number of writhings at both doses. In addition, in the formalin test, ripI reduced the paw licking time at both phases of the test. The effect of the highest dose of ripI in mice formalin test on the early phase was not reversed by naloxone (opioid receptor antagonist) but it was reversed by l-arginine (a nitric oxide precursor) in the late phase, suggesting that ripI may not act through opioid system and possibly acts through inhibition of nitric oxide pathway. In the hot-plate test, ripI increased the reaction time in the hot-plate test at the dose of 25 mg/kg, i.p., confirming the result found in the formalin test. Based on the obtained results, it is suggested that ripI presents antinociceptive activity that may be due to peripheral mechanisms (nitric oxide pathway) and central mechanisms, discarding the involvement of opioid system. PMID- 19557400 TI - Improved quality in the hospital discharge summary reduces medication errors- LIMM: Landskrona Integrated Medicines Management. AB - PURPOSE: We have developed a model for integrated medicines management, including tools and activities for medication reconciliation and medication review. In this study, we focus on improving the quality of the discharge summary including the medication report to reduce medication errors in the transition from hospital to primary and community care. METHODS: This study is a longitudinal study with an intervention group and a control group. The intervention group comprised 52 patients, who were included from 1 March 2006 until 31 December 2006, with a break during summer. Inclusion in the control group was performed in the same wards during the period 1 September 2005 until 20 December 2005, and 63 patients were included in the control group. In order to improve the quality of the medication report, clinical pharmacists reviewed and gave feedback to the physician on the discharge summary before patient discharge, using a structured checklist. Medication errors were then identified by comparing the medication list in the discharge summary with the first medication list used in the community health care after the patient had returned home. RESULTS: By improving the quality of the discharge summary, patients had on average 45% fewer medication errors per patient (P = 0.012). The proportion of patients without medication errors was 63.5% in the control group and 73.1% in the intervention group. However, this increase was not significant (P = 0.319). Patients who used a specific medication dispensing system (ApoDos) had a 5.9-fold higher risk of suffering from medication errors than those without this medication dispensing system (P < 0.001). CONCLUSION: Review and feedback on errors in the discharge summary, including the medication report and a correct medication list, reduced medication errors during the transfer of information from hospital to primary and community care. PMID- 19557401 TI - Pharmacokinetics and dose requirements of factor VIII over the age range 3-74 years: a population analysis based on 50 patients with long-term prophylactic treatment for haemophilia A. AB - PURPOSE: The three aims of this investigation were (1) to develop a population pharmacokinetic (PK) model for factor VIII (FVIII) in haemophilia A patients, with estimates of inter-occasion and inter-individual variance, (2) to investigate whether appropriate dosing of FVIII for regular prophylaxis can be calculated according to patient characteristics, and (3) to present dosing recommendations for initiating prophylactic treatment. METHODS: A population PK model was developed using data from four PK studies on patients aged 7-74 years. The model was tested on sparse FVIII data from 42 outpatient visits by haemophilia prophylaxis patients aged 3-66 years. Dose requirements for prophylaxis were calculated both according to the population model and from empirical Bayesian estimates of FVIII PK in the individual patients. RESULTS: The study data were well characterised by a two-compartment PK model. Body weight, age and type of FVIII preparation (plasma-derived or recombinant) were identified as significant covariates. Inter-occasion variance was lower than inter individual variance for both clearance and volume of the central compartment. The model could reasonably predict FVIII PK in the sparse clinical data. Model predicted doses (based on age and body weight) to maintain a recommended 0.01 U/mL trough level of FVIII with administration on alternate days started at around 60 U/kg in the small children, decreasing to 10 U/kg or less in middle age. However, "true" dose requirements, as estimated from individual PK parameter data, showed a much greater variation. CONCLUSION: Appropriate dosing of FVIII for prophylactic treatment cannot be calculated only from body weight and/or age. However, plausible starting doses for most patients would be 1,000 U every other day. FVIII levels should then be checked for dose adjustment. PMID- 19557402 TI - Endovascular treatment for pediatric intracranial aneurysms. AB - INTRODUCTION: The purpose of this study is to report the characteristics and outcomes of pediatric patients with intracranial aneurysms. METHODS: From 1998 to 2005, 25 pediatric patients (aged < or =17 years) with intracranial aneurysm were treated at our institute. Eleven of 25 patients had subarachnoid hemorrhage. In ten patients, the aneurysm was an incidental finding. One patient presented with cranial nerves dysfunction and three with neurological deficits. The locations of the aneurysms were as follows: vertebral artery (VA; n = 9), middle cerebral artery (MCA; n = 5), posterior cerebral artery (PCA; n = 4), basilar artery (BA; n = 2), anterior communicating artery (n = 2), anterior cerebral artery (n = 2), and internal carotid artery (n = 1). RESULTS: Five patients were treated with selective embolization with coils. Sixteen patients were treated with parent vessel occlusion (PVO). Eight PVOs were performed with balloons and eight were performed with coils. One patient with a VA aneurysm was spontaneously thrombosed 4 days after the initial diagnostic angiogram. In three patients treated with stent alone or stent-assisted coiling, one with BA trunk aneurysm died. One aneurismal recurrence occurred and was retreated. At a mean follow-up duration of 23.5 months, 96% of patients had a Glasgow Outcome Scale score of 4 or 5. CONCLUSIONS: Pediatric intracranial aneurysms occur more commonly in male patients and have a predilection for the VA, PCA, and MCA. PVO is an effective and safe treatment for fusiform aneurysms. Basilar trunk fusiform aneurysms were difficult to treat and were associated with a high mortality rate. PMID- 19557399 TI - Inhaled corticosteroids as combination therapy with beta-adrenergic agonists in airways disease: present and future. AB - Inhaled corticosteroid (ICS) therapy in combination with long-acting beta adrenergic agonists represents the most important treatment for chronic airways diseases such as asthma and chronic obstructive pulmonary disease (COPD). ICS therapy forms the basis for treatment of asthma of all severities, improving asthma control, lung function and preventing exacerbations of disease. Use of ICS has also been established in the treatment of COPD, particularly symptomatic patients, who experience useful gains in quality of life, likely from an improvement in symptoms such as breathlessness and in reduction in exacerbations, and an attenuation of the yearly rate of deterioration in lung function. The addition of long-acting beta-agonist (LABA) therapy with ICS increases the efficacy of ICS effects in moderate-to-severe asthma. Thus, a 800 mug daily dose of the ICS budesonide reduced severe exacerbation rates by 49% compared to a low dose of 200 mug daily, and addition of the LABA formoterol to budesonide (800 mug) led to a 63% reduction. In COPD, the effects of ICS are less prominent but there are beneficial effects on the decline in FEV(1) and the rate of exacerbations. A reduction in the rate of decline in FEV(1) of 16 ml/year with a 25% reduction in exacerbation rate has been reported with the salmeterol and fluticasone combination. A non-significant 17.5% reduction in all-cause mortality rate with ICS and LABA is reported. Chronic inflammation is a feature of both asthma and COPD, although there are site and characteristic differences. ICS targets this inflammation although this effect of ICS is less effective in patients with severe asthma and with COPD; however, addition of LABA may potentiate the anti-inflammatory effects of ICS. An important consideration is the presence of corticosteroid insensitivity in these patients. Currently available ICS have variably potent binding activities to specific glucocorticoid receptors, leading to inhibition of gene expression by either binding to DNA and inducing anti-inflammatory genes or by repressing the induction of pro inflammatory mediators. Local side effects of ICS include oral candidiasis, hoarseness and dysphonia, while systemic side effects, such as easy bruising and reduction in growth velocity or bone mineral densitometry, are usually restricted to doses above maximally recommended doses. Use of LABA alone in patients with asthma increases the risk of asthma-related events including deaths, but this is less observed with the combination of ICS and LABA. Therefore, use of LABA alone is not recommended for asthma therapy. Future progress in ICS development will be characterised by the introduction of ICS with greater efficacy with a limited side-effect profile, and by longer-acting ICS that can be used in combination with once-daily LABAs. Other agents that could improve the efficacy of corticosteroids or reverse corticosteroid insensitivity may be added to ICS. ICS in combination with LABAs will continue to remain the main focus of treatment of airways diseases. PMID- 19557403 TI - Symptomatic hepatic cyst in a child: treatment with single-shot injection of tetracycline hydrochloride. AB - The prevalence of hepatic cysts is 0.1% to 0.5% based on autopsy studies, and 2.5% based on US examinations. Percutaneous therapies are a new alternative to surgery. They include simple percutaneous aspiration, catheter drainage alone, and catheter drainage with sclerotherapy. We present an 11-year-old boy admitted to hospital because of abdominal pain. A diagnosis of simple hepatic cyst was made, which was treated with aspiration and tetracycline hydrochloride solution (5%) injection into the cystic cavity. Complete regression was seen on US and MRI examination at 3 months, with total collapse and deflation of the cyst. The cyst regressed totally, leaving a hyperechoic linear scar on US examination at 1 year. On the basis of the clinical and imaging results obtained, percutaneous sclerotherapy of hepatic cysts can be recommended as the treatment of choice and as a valid alternative to laparoscopy in children. PMID- 19557404 TI - Improving health literacy: informed decision-making rather than informed consent for CT scans in children. PMID- 19557405 TI - What physicians think about the need for informed consent for communicating the risk of cancer from low-dose radiation. AB - BACKGROUND: The National Institute of Environmental Health Sciences, a subsidiary of the Food and Drug Administration, has declared that X-ray radiation at low doses is a human carcinogen. OBJECTIVE: The purpose of our study was to determine if informed consent should be obtained for communicating the risk of radiation induced cancer from radiation-based imaging. MATERIALS AND METHODS: Institutional review board approval was obtained for the prospective survey of 456 physicians affiliated with three tertiary hospitals by means of a written questionnaire. Physicians were asked to state their subspecialty, number of years in practice, frequency of referral for CT scanning, level of awareness about the risk of radiation-induced cancer associated with CT, knowledge of whether such information is provided to patients undergoing CT, and opinions about the need for obtaining informed consent as well as who should provide information about the radiation-induced cancer risk to patients. Physicians were also asked to specify their preference among different formats of informed consent for communicating the potential risk of radiation-induced cancer. Statistical analyses were performed using the chi-squared test. RESULTS: Most physicians stated that informed consent should be obtained from patients undergoing radiation-based imaging (71.3%, 325/456) and the radiology department should provide information about the risk of radiation-induced cancer to these patients (54.6%, 249/456). The informed consent format that most physicians agreed with included modifications to the National Institute of Environmental Health Services report on cancer risk from low-dose radiation (20.2%, 92/456) or included information on the risk of cancer from background radiation compared to that from low-dose radiation (39.5%, 180/456). CONCLUSION: Most physicians do not know if patients are informed about cancer risk from radiation-based imaging in their institutions. However, they believe that informed consent for communicating the risk of radiation-induced cancer should be obtained from patients undergoing radiation-based imaging. PMID- 19557406 TI - Polymorphism and selection in the major histocompatibility complex DRA and DQA genes in the family Equidae. AB - The major histocompatibility complex genes coding for antigen binding and presenting molecules are the most polymorphic genes in the vertebrate genome. We studied the DRA and DQA gene polymorphism of the family Equidae. In addition to 11 previously reported DRA and 24 DQA alleles, six new DRA sequences and 13 new DQA alleles were identified in the genus Equus. Phylogenetic analysis of both DRA and DQA sequences provided evidence for trans-species polymorphism in the family Equidae. The phylogenetic trees differed from species relationships defined by standard taxonomy of Equidae and from trees based on mitochondrial or neutral gene sequence data. Analysis of selection showed differences between the less variable DRA and more variable DQA genes. DRA alleles were more often shared by more species. The DQA sequences analysed showed strong amongst-species positive selection; the selected amino acid positions mostly corresponded to selected positions in rodent and human DQA genes. PMID- 19557407 TI - Efficient production of active Vibrio proteolyticus aminopeptidase in Escherichia coli by co-expression with engineered vibriolysin. AB - The Vibrio proteolyticus aminopeptidase is synthesized as a preproprotein and then converted into an active enzyme by cleavage of the N-terminal propeptide. In recombinant Escherichia coli, however, the aminopeptidase is not processed correctly and the less-active form that has the N-terminal propeptide accumulates in the culture medium. Recently, we isolated a novel vibriolysin that was expressed as an active form in E. coli by random mutagenesis; this enzyme shows potential as a candidate enzyme for the processing of aminopeptidase. The E. coli cells were engineered to co-express the novel vibriolysin along with aminopeptidase. Co-expression of vibriolysin resulted in an approximately 13-fold increase in aminopeptidase activity, and a further increase was observed in the form lacking its C-terminal propeptide. The active aminopeptidase was purified from the culture supernatant including the recombinant vibriolysin by heat treatment and ion exchange and hydroxyapatite chromatography with high purity and 35% recovery rate. This purified aminopeptidase effectively converted methionyl human growth hormone (Met-hGH) to hGH. Thus, this co-expression system provides an efficient method for producing active recombinant V. proteolyticus aminopeptidase. PMID- 19557408 TI - Heterologous expression and characterization of CpI, OcpA, and novel serine-type carboxypeptidase OcpB from Aspergillus oryzae. AB - In the genome of Aspergillus oryzae, 12 genes have been predicted to encode serine-type carboxypeptidases. However, the carboxypeptidase activities of the proteins encoded by these genes have not yet been confirmed experimentally. In this study, we have constructed three of these 12 genes overexpressing strains using Aspergillus nidulans and characterized their overproduced recombinant proteins. Of these three genes, one was previously named cpI; the other two have not been reported yet, and hence, we named them ocpA and ocpB. The recombinant proteins released amino acid residues from the C terminus of peptides, and the activity of the enzymes was inhibited by phenylmethylsulfonyl fluoride, indicating the enzymes to be serine-type carboxypeptidases. Recombinant OcpA, OcpB, and CpI were stable at 45 degrees C, 55 degrees C, and 55 degrees C, respectively, at a low pH. The enzymatic properties of recombinant OcpB were different from those of any reported serine-type carboxypeptidase. On the other hand, recombinant OcpA had similar enzymatic properties to A. oryzae carboxypeptidases O1 and O2. The DNA and N-terminal amino acid sequences of carboxypeptidases O1 and O2 from A. oryzae IAM2640 were similar to those of OcpA. Result of transcriptional analysis of ocpA, ocpB, and cpI suggest differences in transcriptional regulation between these genes. PMID- 19557409 TI - A new non-hydrophobic cell wall protein (CWP10) of Metarhizium anisopliae enhances conidial hydrophobicity when expressed in Beauveria bassiana. AB - A cell wall protein, CWP10, resolved from the conidial formic acid extract of a Metarhizium anisopliae isolate, was characterized as a new 9.9-kDa protein with a 32-aa signal peptide with a central hydrophobic region (ca. 10 residues) at its N terminus. This protein was proven neither to be hydrophobic nor glycosylated and encoded by a 363-bp, single-copy gene with three introns. CWP10 was existent in the conidial extracts of seven of 18 tested M. anisopliae isolates and much more abundant (immunogold-labeled) on conidial walls than in cytoplasm. Integrating the gene into a CWP10-absent strain of Beauveria bassiana led to excellent expression of CWP10 in aerial conidia, increasing net conidial hydrophobicity by 10.8% or adhesion to hydrophobic Teflon by 1.3-fold. However, the expressed protein had no effect on conidial tolerance to thermal and ultraviolet stresses. This is the first report on a non-hydrophobic cell-wall protein enhancing conidial hydrophobicity and adhesion of the fungal species. PMID- 19557410 TI - Magnetic resonance appearance of posterosuperior labral peel back during humeral abduction and external rotation. AB - OBJECTIVE: To describe the magnetic resonance appearance of posterosuperior labral peel back and determine the reliability of MR in the abducted and externally rotated (ABER) position for the prospective diagnosis of arthroscopically proven cases of posterosuperior labral peel back. METHODS: After approval by the institutional review board (IRB) of the University of Pittsburgh Medical Center, USA, databases of patients who underwent arthroscopy over a 2 year period for one of three clinical diagnoses [suspected type 2 superior labrum anterior to posterior (SLAP) tears, posterior instability, or multidirectional instability] were reviewed after anonymization by an honest broker. Sixty-three cases were selected by the following inclusion criteria: operative report documenting labral peel back in the ABER position, age <40 years, and preceding MR arthrogram evaluations with images in the ABER position (n=34). Inclusion criteria for the control group differed from those for the case group insofar as the operative note documented the absence of posterosuperior labral peel back (n=29). Cases and controls were randomized in one list and evaluated independently by two fellowship-trained musculoskeletal radiologists unaware of the surgical results and using a three-point grading system (0 = posterosuperior labrum normally positioned lateral/craniad to glenoid articular plane in ABER; 1 = posterosuperior labral tissue flush with the glenoid articular plane in ABER; 2 = posterosuperior labral tissue identified medial/caudal to glenoid articular plane in ABER). Only one image in ABER showing abnormal posterosuperior labral position was required for a grade of 1 or 2 to be assigned. Sensitivity, specificity, and positive and negative predictive value were calculated as well as the level of agreement between readers (kappa). RESULTS: Both readers assigned a grade of 2 to 25 of 34 patients with surgically proven labral peel back. Of the patients with surgically proven SLAP tears with peel back in ABER, reader A assigned a grade of 1 to seven patients and a grade of 0 to two patients, while reader B assigned a grade of 1 to eight patients and a grade of 0 to one patient. In the control group of 29 patients, reader A assigned 28 patients a grade of 0, one patient a grade 1, and no patients a grade 2. Reader B assigned 27 patients a grade of 0, two a grade 1, and no patients a grade 2. After the data had been dichotomized, with grade 1 and 0 cases both being regarded as negative, the MR criteria showed a sensitivity of 73%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 78%. The kappa coefficient of inter-rater agreement was excellent at 0.9, with disagreement in only four of 63 cases. In five of the 34 cases with peel back, a labral tear, defined by imbibition of contrast agent within a gap between labrum and underlying glenoid bone, could not be identified in standard planes in the neutral position. CONCLUSIONS: The use of the glenoid articular plane as a reference line to evaluate labral peel back in the abducted and externally rotated position is a fairly accurate and highly precise method for detection of posterosuperior labral peel back. Raising the possibility of labral peel back may help alert the arthroscopist to the presence of superior labral instability while the arm is abducted and externally rotated. PMID- 19557411 TI - Darwinian molecular imaging. PMID- 19557412 TI - Uveal melanoma cell-based vaccines express MHC II molecules that traffic via the endocytic and secretory pathways and activate CD8+ cytotoxic, tumor-specific T cells. AB - We are exploring cell-based vaccines as a treatment for the 50% of patients with large primary uveal melanomas who develop lethal metastatic disease. MHC II uveal melanoma vaccines are MHC class I(+) uveal melanoma cells transduced with CD80 genes and MHC II genes syngeneic to the recipient. Previous studies demonstrated that the vaccines activate tumor-specific CD4(+) T cells from patients with metastatic uveal melanoma. We have hypothesized that vaccine potency is due to the absence of the MHC II-associated invariant chain (Ii). In the absence of Ii, newly synthesized MHC II molecules traffic intracellularly via a non-traditional pathway where they encounter and bind novel tumor peptides. Using confocal microscopy, we now confirm this hypothesis and demonstrate that MHC II molecules are present in both the endosomal and secretory pathways in vaccine cells. We also demonstrate that uveal melanoma MHC II vaccines activate uveal melanoma specific, cytolytic CD8(+) T cells that do not lyse normal fibroblasts or other tumor cells. Surprisingly, the CD8(+) T cells are cytolytic for HLA-A syngeneic and MHC I-mismatched uveal melanomas. Collectively, these studies demonstrate that MHC II uveal melanoma vaccines are potent activators of tumor-specific CD4(+) and CD8(+) T cells and suggest that the non-conventional intracellular trafficking pattern of MHC II may contribute to their enhanced immunogenicity. Since MHC I compatibility is unnecessary for the activation of cytolytic CD8(+) T cells, the vaccines could be used in uveal melanoma patients without regard to MHC I genotype. PMID- 19557413 TI - On the origin of serum CD26 and its altered concentration in cancer patients. AB - Dipeptidyl peptidase IV (DPP-IV), assigned to the CD26 cluster, is expressed on epithelial cells and lymphocytes and is a multifunctional or pleiotropic protein. Its peptidase activity causes degradation of many biologically active peptides, e.g. some incretins secreted by the enteroendocrine system. DPP-IV has, therefore, become a novel therapeutic target for inhibitors that extend endogenously produced insulin half-life in diabetics, and several reviews have appeared in recent months concerning the clinical significance of CD26/DPP-IV. Biological fluids contain relatively high levels of soluble CD26 (sCD26). The physiological role of sCD26 and its relation, if any, to CD26 functions, remain poorly understood because whether the process for CD26 secretion and/or shedding from cell membranes is regulated or not is not known. Liver epithelium and lymphocytes are often cited as the most likely source of sCD26. It is important to establish which tissue or organ is the protein source as well as the circumstances that can provoke an abnormal presence/absence or altered levels in many diseases including cancer, so that sCD26 can be validated as a clinical marker or a therapeutic target. For example, we have previously reported low levels of sCD26 in the blood of colorectal cancer patients, which indicated the potential usefulness of the protein as a biomarker for this cancer in early diagnosis, monitoring and prognosis. Through this review, we envisage a role for sCD26 and the alteration of normal peptidase capacity (in clipping enteroendocrine or other peptides) in the complex crosstalk between the lymphoid lineage and, at least, some malignant tumours. PMID- 19557414 TI - Time optimal control of an additional food provided predator-prey system with applications to pest management and biological conservation. AB - Use of additional food has been widely recognized by experimental scientists as one of the important tools for biological control such as species conservation and pest management. The quality and quantity of additional food supplied to the predators is known to play a vital role in the controllability of the system. The present study is continuation of a previous work that highlights the importance of quality and quantity of the additional food in the dynamics of a predator-prey system in the context of biological control. In this article the controllability of the predator-prey system is analyzed by considering inverse of quality of the additional food as the control variable. Control strategies are offered to steer the system from a given initial state to a required terminal state in a minimum time by formulating Mayer problem of optimal control. It is observed that an optimal strategy is a combination of bang-bang controls and could involve multiple switches. Properties of optimal paths are derived using necessary conditions for Mayer problem. In the light of the results evolved in this work it is possible to eradicate the prey from the eco-system in the minimum time by providing the predator with high quality additional food, which is relevant in the pest management. In the perspective of biological conservation this study highlights the possibilities to drive the state to an admissible interior equilibrium (irrespective of its stability nature) of the system in a minimum time. PMID- 19557415 TI - A mathematical model of the sleep/wake cycle. AB - We present a biologically-based mathematical model that accounts for several features of the human sleep/wake cycle. These features include the timing of sleep and wakefulness under normal and sleep-deprived conditions, ultradian rhythms, more frequent switching between sleep and wakefulness due to the loss of orexin and the circadian dependence of several sleep measures. The model demonstrates how these features depend on interactions between a circadian pacemaker and a sleep homeostat and provides a biological basis for the two process model for sleep regulation. The model is based on previous "flip-flop" conceptual models for sleep/wake and REM/NREM and we explore whether the neuronal components in these flip-flop models, with the inclusion of a sleep-homeostatic process and the circadian pacemaker, are sufficient to account for the features of the sleep/wake cycle listed above. The model is minimal in the sense that, besides the sleep homeostat and constant cortical drives, the model includes only those nuclei described in the flip-flop models. Each of the cell groups is modeled by at most two differential equations for the evolution of the total population activity, and the synaptic connections are consistent with those described in the flip-flop models. A detailed analysis of the model leads to an understanding of the mathematical mechanisms, as well as insights into the biological mechanisms, underlying sleep/wake dynamics. PMID- 19557417 TI - [Vascular lesion of the conjunctiva]. AB - A 44-year-old male patient first noticed a pain-free light red swelling of the right eye in 1983. Based on the results of the early indocyanine green angiography, MRI and sonography, a racemose hemangioma of the conjunctiva was diagnosed. Although a racemose hemangioma already exists at birth, it remains clinically silent for a long time and symptoms often first appear in early adulthood. In this patient external surgical excision was terminated due to substantial bleeding. A symptomatic treatment in the sense of surface care is considered to be the best option. PMID- 19557416 TI - Percutaneous radiofrequency ablation of hepatocellular carcinoma abutting the diaphragm and gastrointestinal tracts with the use of artificial ascites: safety and technical efficacy in 143 patients. AB - The purpose of this study was to assess the feasibility, safety and efficacy of radiofrequency ablation (RFA) with the use of artificial ascites for hepatocellular carcinoma (HCC) adjacent to the diaphragm and gastrointestinal tract. One hundred forty-three patients with 181 HCCs who underwent US-guided percutaneous RFA with the use of artificial ascites were retrospectively reviewed. Among the 181 HCCs, 148 HCCs were defined as problematic nodules for two major reasons: poor sonic window or possible thermal injury. We artificially induced ascites before performing RFA by dripping 5% dextrose in a water solution. We assessed the technical success of introducing artificial ascites, technical feasibility of the use of artificial ascites and complications. The technical success rate, as well as the primary and secondary technique success rate, was assessed by regular follow-up CT examinations. RFA with artificial ascites was successfully achieved in 130 of 143 patients. The primary technique effectiveness was 85.3%. During follow-up (mean, 20.4 months), remote intrahepatic recurrence occurred in 49 patients and local tumor progression occurred in 15 patients. Three (2.1%) of the 143 patients experienced major complications (hemoperitoneum, lobar infarction and biloma) related to the RFA procedure. The use of artificial ascites is a simple and useful technique to minimize collateral thermal injury and to improve the sonic window. PMID- 19557418 TI - [New aspects in the therapy of neovascular age related macular degeneration. Current position of the Retinological Society, the Germany Ophthalmologic Society and the Professional Union of Eye Doctors of Germany]. PMID- 19557420 TI - Rapidly regrowing lipoma in lipomeningomyelocele: a case report. AB - INTRODUCTION: Although it is widely accepted that lipomas in lipomeningomyelocele (LMMC) are anatomically stable lesions, we present a case of rapidly growing intradural lipoma accompanied by neurological deterioration. An 8-year-old girl with rapidly regrowing LMMC was admitted to our neurosurgical clinic. She was delivered by cesarean section, and at 47 days old, she underwent detethering surgery because of aggravating weakness. We performed her detethering operation immediately rather than at 3 months as had been previously scheduled because of the rapidly growing lipoma. The second operation was performed at age 8 years because of a regrowing intradural lipoma and aggravating leg deformity. Even if LMMC is expected to be surgically corrected at 2 or 3 months after birth, lipomas could grow rapidly beyond expectation. DISCUSSION: Close monitoring and follow-up before and after surgery is needed for optimum surgical timing, in order to avoid deteriorating neurological symptoms. The possibility of the rapid growth of intradural lipomas offers additional support for early prophylactic surgery in patients with LMMC. CONCLUSION: The potential for rapid growth of spinal lipomas should be considered during first few months of life or even long after surgery. Early diagnosis and optimal surgery are still essential for the treatment of patients with LMMC since there is a high likelihood that residual neurological functions can be preserved. PMID- 19557421 TI - Intraventricular and subarachnoid basal cisterns neurocysticercosis: a comparative study between traditional treatment versus neuroendoscopic surgery. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of traditional treatment and minimal invasive flexible endoscopy surgery (MIFNES) in the treatment of intraventricular and subarachnoid basal cisterns neurocysticercosis (NCC). METHODS: This was an observational comparative study of two independent series with a total of 140 patients with extremely severe forms of NCC from two different institutions. All 83 patients submitted for traditional treatment series received albendazole, and some of them received additionally praziquantel. Each cycle of both regimens lasted 4 weeks. The majority of these patients had at least one ventriculoperitoneal (VP) shunt. The rest 57 patients were submitted to the MIFNES treatment. The follow-up period was at least 6 months. RESULTS: In all patients of both series cysticercal cysts disappeared, became calcified, or were removed. Symptoms of 136 patients improved. Four patients died. The average in the quality of life measured using the Karnofsky scale improved from a mean of 52.22 and 52.44 at the beginning to 85.48 and 90.37 at 6 months (p < 0.003), in the traditional treatment and MIFNES series, respectively. From traditional treatment, almost all patients remained with at least one VP shunt, and from the MIFNES series only 12 patients. CONCLUSIONS: The authors postulate that MIFNES is a good alternative for the management of intraventricular and subarachnoid basal cisterns NCC because it allows removal of most of the parasites, rapid recovery of the patients, and removal and placement of shunt under direct vision when necessary. Traditional treatment is a second option where the MIFNES procedure is not available. PMID- 19557422 TI - [Gerontorheumatology. Aspects of diagnosis, course and therapy of inflammatory rheumatic disease in advanced age]. AB - Gerontorheumatology deals with the particular features of onset, course and treatment of rheumatic diseases in patients of advanced age. The initial diagnosis of inflammatory rheumatic disease after the age of 60 is hindered by the frequency of non-specific general disease symptoms. The most important gerontorheumatological diseases include rheumatoid arthritis first occurring at advanced age ("late onset rheumatoid arthritis", LORA), rheumatic polymyalgia and giant cell arteritis. Important differential diagnoses in older rheumatology patients are RS3PE syndrome, polyarticular chondrocalcinosis and paraneoplastic rheumatic syndrome. In principle, all specific basic therapeutics, immunosuppressants and biologics can be used for the therapy of these diseases. However, careful dose selection is particularly necessary in the initial phase, taking age-specific limitations in organ function and metabolism into consideration; moreover, close-meshed tolerance tests should be carried out. PMID- 19557423 TI - p62/SQSTM1 is overexpressed and prominently accumulated in inclusions of sporadic inclusion-body myositis muscle fibers, and can help differentiating it from polymyositis and dermatomyositis. AB - p62, also known as sequestosome1, is a shuttle protein transporting polyubiquitinated proteins for both the proteasomal and lysosomal degradation. p62 is an integral component of inclusions in brains of various neurodegenerative disorders, including Alzheimer disease (AD) neurofibrillary tangles (NFTs) and Lewy bodies in Parkinson disease. In AD brain, the p62 localized in NFTs is associated with phosphorylated tau (p-tau). Sporadic inclusion-body myositis (s IBM) is the most common progressive muscle disease associated with aging, and its muscle tissue has several phenotypic similarities to AD brain. Abnormal accumulation of intracellular multiprotein inclusions, containing p-tau in the form of paired helical filaments, amyloid-beta, and several other "Alzheimer characteristic proteins", is a characteristic feature of the s-IBM muscle fiber phenotype. Diminished proteasomal and lysosomal protein degradation appear to play an important role in the formation of intra-muscle-fiber inclusions. We now report that: (1) in s-IBM muscle fibers, p62 protein is increased on both the protein and the mRNA levels, and it is strongly accumulated within, and as a dense peripheral shell surrounding, p-tau containing inclusions, by both the light- and electron-microscopy. Accordingly, our studies provide a new, reliable, and simple molecular marker of p-tau inclusions in s-IBM muscle fibers. The prominent p62 immunohistochemical positivity and pattern diagnostically distinguish s-IBM from polymyositis and dermatomyositis. (2) In normal cultured human muscle fibers, experimental inhibition of either proteasomal or lysosomal protein degradation caused substantial increase of p62, suggesting that similar in vivo mechanisms might contribute to the p62 increase in s-IBM muscle fibers. PMID- 19557424 TI - Cocktail therapy for femoral head necrosis of the hip. AB - INTRODUCTION: Cocktail therapy consists of extracorporeal shock wave treatment (ESWT), hyperbaric oxygen therapy (HBO) and alendronate. We hypothesized that this combined therapy may produce better results than ESWT alone when treating hip necrosis. METHODS: This prospective, randomized study evaluated the effectiveness of cocktail therapy in early hip necrosis, and then compared it to shock wave treatment alone. Sixty-three patients (98 hips) were randomly divided into two groups. Group A consisted of 28 patients (50 hips) who received cocktail therapy, and group B consisted of 35 patients (48 hips) who received only ESWT. Each affected hip was treated with 6,000 shocks of ESWT at 28 kV (equivalent to 0.62 mJ/mm2) in a single session. Cocktail patients also received HBO therapy performed in a multi-place chamber daily for 20 treatments, and they were administered 70 mg of alendronate orally per week for 1 year. The evaluations included clinical assessment, radiograph and MRI. RESULTS: After a minimum of 2 years of follow-up, the overall results showed 74% improved, 16% unchanged and 10% worsened in group A; and 79.2% improved, 10.4% unchanged and 10.4% worsened in group B (P = 0.717). Total hip replacement (THR) was performed for 10% of group A and 10.4% of group B (P = 0.946). MRI showed significant reduction in bone marrow edema and a trend of decrease in the size of the lesions in both groups, however, no difference was noted between the two groups. Cocktail therapy is effective for early hip necrosis, but the results are comparable to ESWT in short-term follow up. The joint effects of HBO and alendronate over EWST were not observed. CONCLUSION: Long-term results are needed to determine whether or not the synergistic effects of ESWT, HBO and alendronate over ESWT alone exist in early hip necrosis. PMID- 19557425 TI - ESR dosimetry study on population of settlements nearby Ust-Kamenogorsk city, Kazakhstan. AB - The method of electron spin resonance (ESR) dosimetry has been applied to human tooth enamel, to obtain individual absorbed doses of residents of settlements in vicinity of Ust-Kamenogorsk city, Kazakhstan (located about 400 km to the east from the epicenter of explosion at the Semipalatinsk Nuclear Test Site, SNTS). This region developed as a major mining and metallurgical center during the Soviet period (uranium production). Most of the investigated settlements (Ust Kamenogorsk city, Glubokoe, Tavriya, Gagarino) are located near the central axis of the radioactive fallout trace that originated from the surface nuclear test on 24 August 1956, while the Kokpekty settlement (located 400 km to the Southeast from SNTS) was chosen as a control because it was not subjected to any radioactive contamination. In total, 44 samples were measured. It was found that the excess doses obtained after subtraction of natural background radiation ranged up to about 114 mGy for residents of Ust-Kamenogorsk city, whose tooth enamel was formed before 1956. For residents of Gagarino, excess doses did not exceed 47 mGy for all ages. For residents of Tavriya, the maximum excess dose was 54 mGy, while for residents of Glubokoe it was about 58 mGy. For the population of the Shemonaikha settlements located at a distance of about 70 km from the central axis of the radioactive fallout trace, highest excess doses were 110 mGy. These high doses may be due to the influence of uranium enterprises located in that region, but probably not due to dental X-ray irradiation. For a final conclusion on the radiological situation in this region, the number of samples was too small and, therefore, more work is required to obtain representative results. PMID- 19557426 TI - Synaptonemal complex assembly and H3K4Me3 demethylation determine DIDO3 localization in meiosis. AB - Synapsis of homologous chromosomes is a key meiotic event, mediated by a large proteinaceous structure termed the synaptonemal complex. Here, we describe a role in meiosis for the murine death-inducer obliterator (Dido) gene. The Dido gene codes for three proteins that recognize trimethylated histone H3 lysine 4 through their amino-terminal plant homeodomain domain. DIDO3, the largest of the three isoforms, localizes to the central region of the synaptonemal complex in germ cells. DIDO3 follows the distribution of the central region protein SYCP1 in Sycp3-/- spermatocytes, which lack the axial elements of the synaptonemal complex. This indicates that synapsis is a requirement for DIDO3 incorporation. Interestingly, DIDO3 is missing from the synaptonemal complex in Atm mutant spermatocytes, which form synapses but show persistent trimethylation of histone H3 lysine 4. In order to further address a role of epigenetic modifications in DIDO3 localization, we made a mutant of the Dido gene that produces a truncated DIDO3 protein. This truncated protein, which lacks the histone-binding domain, is incorporated in the synaptonemal complex irrespective of histone trimethylation status. DIDO3 protein truncation in Dido mutant mice causes mild meiotic defects, visible as gaps in the synaptonemal complex, but allows for normal meiotic progression. Our results indicate that histone H3 lysine 4 demethylation modulates DIDO3 localization in meiosis and suggest epigenetic regulation of the synaptonemal complex. PMID- 19557427 TI - Quantitative electron microscopy shows uniform incorporation of triglycerides into existing lipid droplets. AB - The lipid droplet (LD) is an organelle with a lipid ester core and a surface phospholipid monolayer. The mechanism of LD biogenesis is not well understood. The present study aimed to elucidate the LD growth process, for which we developed a new electron microscopic method that quantifies the proportion of existing and newly synthesized triglycerides in individual LDs. Our method takes advantage of the reactivity of unsaturated fatty acids and osmium tetroxide, which imparts LDs an electron density that reflects fatty acid composition. With this method, existing triglyceride-rich LDs in 3Y1 fibroblasts were observed to incorporate newly synthesized triglycerides at a highly uniform rate. This uniformity and its persistence even after microtubules were depolymerized suggest that triglycerides in fibroblasts are synthesized in the local vicinity of individual LDs and then incorporated. In contrast, LDs in 3T3-L1 adipocytes showed heterogeneity in the rate at which lipid esters were incorporated, indicating different mechanisms of LD growth in fibroblasts and adipocytes. PMID- 19557428 TI - The two-pore channel TPCN2 mediates NAADP-dependent Ca(2+)-release from lysosomal stores. AB - Second messenger-induced Ca(2+)-release from intracellular stores plays a key role in a multitude of physiological processes. In addition to 1,4,5-inositol trisphosphate (IP(3)), Ca(2+), and cyclic ADP ribose (cADPR) that trigger Ca(2+) release from the endoplasmatic reticulum (ER), nicotinic acid adenine dinucleotide phosphate (NAADP) has been identified as a cellular metabolite that mediates Ca(2+)-release from lysosomal stores. While NAADP-induced Ca(2+)-release has been found in many tissues and cell types, the molecular identity of the channel(s) conferring this release remained elusive so far. Here, we show that TPCN2, a novel member of the two-pore cation channel family, displays the basic properties of native NAADP-dependent Ca(2+)-release channels. TPCN2 transcripts are widely expressed in the body and encode a lysosomal protein forming homomers. TPCN2 mediates intracellular Ca(2+)-release after activation with low-nanomolar concentrations of NAADP while it is desensitized by micromolar concentrations of this second messenger and is insensitive to the NAADP analog nicotinamide adenine dinucleotide phosphate (NADP). Furthermore, TPCN2-mediated Ca(2+)-release is almost completely abolished when the capacity of lysosomes for storing Ca(2+) is pharmacologically blocked. By contrast, TPCN2-specific Ca(2+)-release is unaffected by emptying ER-based Ca(2+) stores. In conclusion, these findings indicate that TPCN2 is a major component of the long-sought lysosomal NAADP dependent Ca(2+)-release channel. PMID- 19557429 TI - Cadmium decreases crown root number by decreasing endogenous nitric oxide, which is indispensable for crown root primordia initiation in rice seedlings. AB - Cadmium (Cd) is toxic to crown roots (CR), which are essential for maintaining normal growth and development in rice seedlings. Nitric oxide (NO) is an important signaling molecule that plays a pivotal role in plant root organogenesis. Here, the effects of Cd on endogenous NO content and root growth conditions were studied in rice seedlings. Results showed that similar to the NO scavenger, cPTIO, Cd significantly decreased endogenous NO content and CR number in rice seedlings, and these decreases were recoverable with the application of sodium nitroprusside (SNP, a NO donor). Microscopic analysis of root collars revealed that treatment with Cd and cPTIO inhibited CR primordia initiation. In contrast, although SNP partially recovered Cd-caused inhibition of CR elongation, treatment with cPTIO had no effect on CR elongation. L: -NMMA, a widely used nitric oxide synthase (NOS) inhibitor, decreased endogenous NO content and CR number significantly, while tungstate, a nitrate reductase (NR) inhibitor, had no effect on endogenous NO content and CR number. Moreover, enzyme activity assays indicated that treatment with SNP inhibited NOS activity significantly, but had no effect on NR activity. All these results support the conclusions that a critical endogenous NO concentration is indispensable for rice CR primordia initiation rather than elongation, NOS is the main source for endogenous NO generation, and Cd decreases CR number by inhibiting NOS activity and thus decreasing endogenous NO content in rice seedlings. PMID- 19557430 TI - Differentiation patterning of vascular smooth muscle cells (VSMC) in atherosclerosis. AB - To investigate the involvement of transdifferentiation and dedifferentiation phenomena inside atherosclerotic plaques, we analyzed the differentiation status of vascular smooth muscle cells (VSMC) in vitro and in vivo. Forty normal autoptic and 20 atherosclerotic carotid endarterectomy specimens as well as 20 specimens of infrarenal and suprarenal aortae were analyzed for the expression of cytokeratins 7 and 18 and beta-catenin as markers (epithelial transdifferentiation) as well as CD31 and CD34 (embryonic dedifferentiation) by conventional and double fluorescence immunohistochemistry and reverse transcription polymerase chain reaction. Looking at these markers, additional cell culture experiments with human aortic (HA)-VSMC were done under stimulation with IL-1beta, IL-6, and TNF-alpha. Cytokeratins and beta-catenin were expressed significantly higher in atherosclerotic than in normal carotids primarily localized in VSMC of the shoulder/cap region of atherosclerotic lesions. Additionally, heterogeneous cellular coexpression of CD31 and/or CD34 was observed in subregions of progressive atherosclerotic lesions by VSMC. The expression of those differentiation markers by stimulated HA-VSMC showed a time and cytokine dependency in vitro. Our findings show that (1) VSMC of progressive atheromas have the ability of differentiation, (2) that transdifferentiation and dedifferentiation phenomena are topographically diverse localized in the subregions of advanced atherosclerotic lesions, and (3) are influenced by inflammatory cytokines like IL-1beta, IL-6, and TNF-alpha. PMID- 19557431 TI - Protein analysis of tissues--current views and clinical perspectives. AB - Proteomics raises high expectations in finding novel and reliable biomarkers for diagnosis, prognosis and therapy prediction. The goal of the 2-day workshop "Protein analysis of tissues-current views and clinical perspectives" was to bring together scientists from multiple areas of protein research interested in tissue analysis. PMID- 19557433 TI - Spatial distribution, blood feeding pattern, and role of Anopheles funestus complex in malaria transmission in central Kenya. AB - Studies were conducted to determine the role of sibling species of Anopheles funestus complex in malaria transmission in three agro-ecosystems in central Kenya. Mosquitoes were sampled indoors and outdoors, and rDNA PCR was successfully used to identify 340 specimens. Anopheles parensis (91.8%), A. funestus (6.8%), and Anopheles leesoni (1.5%) were the three sibling species identified. A. parensis was the dominant species at all study sites, while 22 of 23 A. funestus were collected in the non-irrigated study site. None of the 362 specimens tested was positive for Plasmodium falciparum circumsporozoite proteins by enzyme-linked immunosorbent assay. The most common blood-meal sources (mixed blood meals included) for A. parensis were goat (54.0%), human (47.6%), and bovine (39.7%), while the few A. funestus s.s. samples had fed mostly on humans. The human blood index (HBI) for A. parensis (mixed blood meals included) in the non-irrigated agro-ecosystem was 0.93 and significantly higher than 0.33 in planned rice agro-ecosystem. The few samples of A. funestus s.s. and A. funestus s.l. also showed a trend of higher HBI in the non-irrigated agro-ecosystem. We conclude that agricultural practices have significant influence on distribution and blood feeding behavior of A. funestus complex. Although none of the species was implicated with malaria transmission, these results may partly explain why non-irrigated agro-ecosystems are associated with higher risk of malaria transmission by this species compared to irrigated agro-ecosystems. PMID- 19557434 TI - Herd factors influencing oocyst production of Eimeria and Cryptosporidium in Estonian dairy cattle. AB - Cryptosporidium and Eimeria are intestinal parasites which are sensitive to the surroundings, behaviour and well-being of their host. In the present study, a range of factors related to farm management systems, environment, housing and herd characteristics were investigated with regard to alterations in oocyst excretion in cattle, using a mixed-effects model. Information and samples for three age categories were obtained from 45 Estonian dairy farms, located in 15 counties. Leaving the calf with the mother after birth reduced the risk of shedding higher levels of Cryptosporidium (OR = 0.20) and Eimeria (OR = 0.68) oocysts in all animals. The calves younger than 3 months kept on farms housing at least 150 animals had less risk (OR = 0.39) of producing higher numbers of Cryptosporidium oocysts. A somewhat lower infection level was observed in 3- to 12-month-old animals housed in separate buildings (OR = 0.64). The chance of shedding higher levels of Eimeria doubled (OR = 2.27) in cattle older than a year in case a vacancy period was used before replacing animals in pens and tripled (OR = 2.94) when the relative humidity exceeded 75% in the cowshed. Winter reduced the odds (OR = 0.25) of shedding Eimeria oocysts in the oldest animals compared to the fall season. Simple changes in handling and housing of cattle may produce a positive effect on controlling coccidian infections in Estonian dairy herds. PMID- 19557432 TI - A predicted functional single-nucleotide polymorphism of bone morphogenetic protein-4 gene affects mRNA expression and shows a significant association with cutaneous melanoma in Southern Italian population. AB - PURPOSE: An altered expression of bone morphogenetic protein 4 (BMP4) has been found in malignant melanoma cells. We performed an association study to investigate the effect of putative functional single nucleotide polymorphisms (SNPs) of BMP4 on development of cutaneous melanoma (CM). METHODS: We selected the predicted functional SNPs 6007 C/T (rs17563) and -3445 T/G (rs4898820) by the combination of three computational tools (FASTSNP, F-SNP and SNP Function Portal) plus another tool (SNP@promoter) skilled in identifying SNPs in transcription regulatory regions. Both SNPs were genotyped in a case-control study of 215 individuals with CM and 342 controls. We also evaluated the BMP4 hypothetical mRNA secondary structure by GeneBee program, the BMP4 mRNA levels and protein concentrations according to the genotype of two selected SNPs in transformed B cells of 80 controls and in plasma samples of 38 controls, respectively. RESULTS: The BMP4 T-allele was associated with CM (OR: 1.39, 95% CI: 1.09-1.78, P = 0.007). The T-allele was predicted to change mRNA structure and the BMP4 mRNA levels were significantly higher in T-allele carriers compared with C-allele carriers (P = 0.01), even the BMP4 protein plasma levels were higher among T allele carries, but without reaching the statistical significance. No significant association was found between the SNP -3445 T/G alleles and either the risk of CM, or the mRNA levels of BMP4. CONCLUSIONS: This study evidences the relevance of using bioinformatics tools in searching for cancer-associated gene polymorphisms and suggests that the predicted functional SNP 6007 C/T affects BMP4 gene expression and the risk to development of CM. PMID- 19557435 TI - Effects of curcumin on Cryptosporidium parvum in vitro. AB - Cryptosporidium parvum is a zoonotic protozoan parasite having peculiarities among the apicomplexa that could be responsible for its resistance to some drugs and disinfectants against coccidia. The awareness of Cryptosporidium as a health problem in man and animal is increasing and potent drugs are urgently needed. Curcumin, a natural polyphenolic compound, has been found to be active against a variety of diseases including anticarcinogenic, antimicrobial, and antiprotozoal effects. We investigated the effects of curcumin on infectivity and development of C. parvum in a recently established in vitro system combining infection of human ileocecal adenocarcinoma cell cultures with quantification of intracellular parasites by quantitative polymerase chain reaction. Curcumin was found to be effective (>95% inhibition of parasite growth) at 50 microM for 24 h when infected cultures were exposed for more than 12 h. Withdrawal of curcumin after 24 h of exposure did not result in a significant resumption of C. parvum growth. The invasion of host cells by sporozoites (infectivity) was found to be inhibited at least 65% in the presence of 200 microM curcumin. No significant reduction of viability of C. parvum oocysts after incubation with curcumin was recorded. Altogether, curcumin showed promising anticryptosporidial effects under in vitro conditions and deserves further exploration. PMID- 19557436 TI - Comparative efficacy of Annona squamosa and Azadirachta indica extracts against Boophilus microplus Izatnagar isolate. AB - In the search of developing herbal acaricides, eight medicinal plants were screened for their efficacy against Boophilus microplus, the widely distributed tick species in India. Of the seven extracts screened, the extracts prepared from the Annona squamosa seed showed very high level of efficacy (70.8%) after 24 h of treatment. The effect of treatment on oviposition of the survived ticks was also assessed, and a significant reduction (P < 0.05) in the reproductive index was noted in comparison to control. When efficacy of the in vitro optimized concentration of A. squamosa was compared with previously tested extract of Azadirachta indica in in vivo model, it was observed that the extracts prepared from A. indica is more efficacious than the extracts of A. squamosa. A comparable efficacy against B. microplus fed on animals treated with herbal extracts and commonly used synthetic acaricide was noted. The possibility of using the herbal extracts in IPM format for the management of ticks is discussed. PMID- 19557437 TI - Surface ultrastructure of Plagiorchis muris growth and developmental stages in rats, the final host. AB - In the excysted metacercaria, the whole fluke surface was covered with peg-shaped tegumental spines. Ciliated sensory papillae (type I) were concentrated around the oral sucker, and non-ciliated sensory papillae (type II) were aligned on the lip of oral and ventral suckers. Several type II papillae were aligned laterally with linear symmetry between the oral and ventral suckers. In juvenile flukes, cytoplasmic processes were band-shaped on the anterior half of the body surface and velvety on the posterior half. In adult flukes, cytoplasmic processes were differentiated into velvety processes, and densities of tegumental spines were reduced on the posterior half of the body. Flukes grew to be elongated and leaf like as adults and retained a surface ultrastructure that was similar to that of juveniles in terms of the distribution of tegumental spines and sensory papillae. From the above results, the authors suppose that the marked differentiation of cystoplasmic processes and the reduced density of tegumental spines observed on the posterior half of the body surface are closely related to the development of reproductive organs therein. PMID- 19557439 TI - Preparation and characterization of epoxy-functionalized magnetic chitosan beads: laccase immobilized for degradation of reactive dyes. AB - Cross-linked magnetic chitosan beads were prepared by phase-inversion technique in the presence of epichlorohydrin under alkaline condition, and used for covalent immobilization of laccase. The activity of the immobilized laccase on the magnetic chitosan was about 260 U (g/dry beads) with an enzyme loading of about 16.33 +/- 0.39 mg [(g/dry beads) mg/g]. Kinetic parameters, V (max) and K (m) values were determined as 21.7 U/mg protein and 9.4 microM for free enzyme, and 15.6 U/mg protein and 19.7 microM for the immobilized laccase, respectively. The operational and thermal stabilities of the immobilized laccase were improved compared to free counterpart. The immobilized laccase was operated in a batch reactor for the decolorization of reactive dyes from aqueous solution. The laccase immobilized on magnetic chitosan beads was very effective for removal of textile dyes from aqueous solution which creates an important environmental problem in the discharged textile dying solutions. PMID- 19557438 TI - Redefined genomic architecture in 15q24 directed by patient deletion/duplication breakpoint mapping. AB - We report four new patients with a submicroscopic deletion in 15q24 manifesting developmental delay, short stature, hypotonia, digital abnormalities, joint laxity, genital abnormalities, and characteristic facial features. These clinical features are shared with six recently reported patients with a 15q24 microdeletion, supporting the notion that this is a recognizable syndrome. We describe a case of an ~2.6 Mb microduplication involving a portion of the minimal deletion critical region in a 15-year-old male with short stature, mild mental retardation, attention deficit hyperactivity disorder, Asperger syndrome, decreased joint mobility, digital abnormalities, and characteristic facial features. Some of these features are shared with a recently reported case with a 15q24 microduplication involving the minimal deletion critical region. We also report two siblings and their mother with duplication adjacent and distal to this region exhibiting mild developmental delay, hypotonia, tapering fingers, characteristic facial features, and prominent ears. The deletion and duplication breakpoints were mapped by array comparative genomic hybridization and the genomic structure in 15q24 was analyzed further. Surprisingly, in addition to the previously recognized three low-copy repeat clusters (BP1, BP2, and BP3), we identified two other paralogous low-copy repeat clusters that likely mediated the formation of alternative sized 15q24 genomic rearrangements via non-allelic homologous recombination. PMID- 19557440 TI - [Is the analgesic effect of acupuncture a placebo effect?]. AB - The German acupuncture trials ART and GERAC have shown that acupuncture and sham or minimal acupuncture were equally effective in the reduction of chronic pain symptoms. These results have prompted an ongoing discussion as to whether acupuncture exerts its effects through a placebo response. Increasing knowledge about the neurobiology of pain and its intrinsic control suggests a combination of acupuncture-specific neurophysiologic effects combined with effects that match those of expectation-induced placebo analgesia. PMID- 19557441 TI - Ectomycorrhizal fungal communities associated with Pinus thunbergii in the eastern coastal pine forests of Korea. AB - We investigated the ectomycorrhizal (ECM) fungal colonization status of Pinus thunbergii mature trees and regenerating seedlings varying in age in coastal pine forests on the east coast of Korea. We established one 20 x 20-m plot at each of two study sites at P. thunbergii coastal forests in Samcheok. Fifty soil blocks (5 x 5 x 15 cm) were sampled at regular intervals, and ten P. thunbergii seedlings of age 0, 1-3, 3-5, and 5-10 years were sampled in each study plot. In total of 27 ECM fungal taxa, Cenococcum geophilum was dominant, followed by Russula sp., Sebacina sp., and unidentified Cortinuris sp. in mature trees. In 0 year-old seedlings, some fungal species such as Sebacina sp., C. geophilum, and unidentified Cortinarius sp. were dominant whereas only C. geophilum was dominant after 1 year, and there were no apparent succession patterns in ECM fungal compositions beyond a host age of 1 year. Most ECM fungal taxa that had colonized seedlings of each age class were also observed in roots of mature trees in each site. These taxa accounted for 86.7-100% and 96.4-98.4% of ECM abundance in seedlings and mature trees, respectively. The results indicate that the species composition of ECM fungal taxa colonizing seedlings of different age in forests is similar to that of surrounding mature trees. Our results also showed that C. geophilum is a common and dominant ECM fungus in P. thunbergii coastal forests and might play a significant role in their regeneration. PMID- 19557442 TI - Effects of compressive loading on biomechanical properties of disc and peripheral tissue in a rat tail model. AB - Intervertebral disc degeneration induced by mechanical compression is an important issue in spinal disorder research. In this study, the biomechanical aspect of the rat tail model was investigated. An external loading device equipped with super-elastic TiNi springs was developed to apply a precise load to the rat tail. By using this device, rat tail discs were subjected to compressive stress of 0.5 or 1.0 MPa for 2 weeks. Discs in the sham group received an attachment of the device but no loading. After the experimental period, first the intact tail with peripheral tissues (PT) such as tendon and skin and then the retrieved disc without PT were subjected to a uniaxial tension-compression test; biomechanical characteristics such as range of motion (ROM), neutral zone (NZ), and hysteresis loss (HL) were evaluated. Furthermore, the load-bearing contribution of PT in the intact tail was estimated by comparing the load displacement curves obtained by the mechanical tests performed with and without PT. The experimental findings revealed that the continuous compressive stress induced reduction in disc thickness. The intact tail demonstrated decreases in ROM and NZ as well as increases in HL. On the other hand, the retrieved disc demonstrated increases in ROM, NZ, and HL. Further, a significant increase in the load-bearing contribution of PT was indicated. These findings suggest that the load-bearing capacity of the disc was seriously deteriorated by the application of compressive stress of 0.5 or 1.0 MPa for 2 weeks. PMID- 19557444 TI - Peck tracking: a method for localizing critical features within complex pictures for pigeons. AB - The pigeon is a standard animal in comparative psychology and is frequently used to investigate visuocognitive functions. Nonetheless, the strategies that pigeons use to discriminate complex visual stimuli remain a difficult area of study. In search of a reliable method to identify features that control the discrimination behaviour, pecking location was tracked using touch screen technology in a people absent/people-present discrimination task. The correct stimuli contained human figures anywhere on the picture, but the birds were not required to peck on that part. However, the stimuli were designed in a way that only the human figures contained distinguishing information. All pigeons focused their pecks on a subarea of the distinctive human figures, namely the heads. Removal of the heads significantly impaired performance, while removal of other distinctive parts did not. Thus, peck tracking reveals the location within a complex visual stimulus that controls discrimination behaviour, and might be a valuable tool to reveal the strategies pigeons apply in visual discrimination tasks. PMID- 19557443 TI - Effect of computer assistance on observer performance of approximal caries diagnosis using intraoral digital radiography. AB - Logicon Caries Detector (LDDC) is the only commercially available computer assisted diagnostic system for caries diagnosis. The object of this study is to elucidate the efficacy of LDDC when used by inexperienced dentists. Fifty extracted teeth were imaged using an RVG6000. Seven dentists who had just passed the Japanese National Dental Board Examination observed those images without LDDC (woLDDC) and assessed the probability that caries lesions were present, then re assessed the same teeth using LDDC (wLDDC). The areas under the receiver operating characteristic curves (Az) were compared. No statistically significant difference was found between woLDDC Az values and wLDDC Az values when caries lesions of all depths were considered. When positive cases were restricted to caries lesions in the inner half of the enamel or to dentine caries lesions, however, wLDDC Az values were significantly larger than woLDDC (p = 0.043 and 0.018, respectively). PMID- 19557445 TI - Flow cytometry applications in the food industry. AB - Flow cytometry has become a valuable tool in food microbiology. By analysing large numbers of cells individually using light-scattering and fluorescence measurements, this technique reveals both cellular characteristics and the levels of cellular components. Flow cytometry has been developed to rapidly enumerate microorganisms; to distinguish between viable, metabolically active and dead cells, which is of great importance in food development and food spoilage; and to detect specific pathogenic microorganisms by conjugating antibodies with fluorochromes, which is of great use in the food industry. In addition, high speed multiparametric data acquisition, analysis and cell sorting, which allow other characteristics of individual cells to be studied, have increased the interest of food microbiologists in this technique. This mini-review gives an overview of the principles of flow cytometry and examples of the application of this technique in the food industry. PMID- 19557446 TI - Exploration of geosmin synthase from Streptomyces peucetius ATCC 27952 by deletion of doxorubicin biosynthetic gene cluster. AB - Thorough investigation of Streptomyces peucetius ATCC 27952 genome revealed a sesquiterpene synthase, named spterp13, which encodes a putative protein of 732 amino acids with significant similarity to S. avermitilis MA-4680 (SAV2163, GeoA) and S. coelicolor A3(2) (SCO6073). The proteins encoded by SAV2163 and SCO6073 produce geosmin in the respective strains. However, the spterp13 gene seemed to be silent in S. peucetius. Deletion of the doxorubicin gene cluster from S. peucetius resulted in increased cell growth rate along with detectable production of geosmin. When we over expressed the spterp13 gene in S. peucetius DM07 under the control of an ermE* promoter, 2.4 +/- 0.4-fold enhanced production of geosmin was observed. PMID- 19557447 TI - Phytotoxicity tests of solid wastes and contaminated soils in the Czech Republic. AB - BACKGROUND, AIM, AND SCOPE: The purpose of this study was to compare the suitability of different phytotoxicity testing procedures for the evaluation of toxicity associated with both soil contamination and solid wastes, both of which can be of environmental risk to plants. Ten different representative types of contaminated soils and solid waste samples were chosen from the Czech Republic. MATERIALS AND METHODS: Both solid-phase and aquatic toxicity testing procedures on mono- and dicotyledonous plants were performed using Lactuca sativa L., Sinapis alba L., Hordeum vulgare L., Triticum aestivum L., Lemna minor L., and the chlorococcal algae Desmodesmus subspicatus (syn. Scenedesmus subspicatus), strain BRINKMANN: 1953/SAG 86.81. An innovative classification scheme, using the intensity of toxic effects upon the plants, is presented in the study. Detailed chemical characterizations of both solid samples and their aquatic elutriates were carried out, using the appropriate ISO guidelines. In the solid samples, all the congeners of polychlorinated biphenyls were analyzed, together with 16 U.S. EPA polyaromatic hydrocarbons, the aggregate of C10-C40 hydrocarbons, total organic carbon, extractable organic halogens, as well as the majority of the environmentally toxic metals. In the aquatic elutriates, parameters analyzed were pH, conductivity, dissolved organic content, phenol index, main anions, and the majority of the environmentally relevant metals. RESULTS: Eight out of ten samples tested expressed phytotoxic properties on tested organisms. Only three of the samples were toxic to both aquatic and terrestrial organisms in the tests. This demonstrates how different substances present in different samples can express different types of toxic effects, resulting in the illogical substituting terrestrial bioassays with aquatic ones. DISCUSSION: Based upon our experience, we propose the following battery of bioassays for use in the characterization of toxic properties of solid wastes and contaminated soils: Aquatic ecosystems were tested by the algae D. subspicatus and plant L. minor; and the terrestrial ecosystems were tested by the dicotyledonous L. sativa and monocotyledonous H. vulgare. This proposed new battery of bioassays for the detection of phytotoxicity of both solid wastes and contaminated soils has higher sensitivity (as well as greater ecological relevance) compared to the battery of bioassays currently used in the Czech Republic. CONCLUSIONS: The tests currently used for regulatory purposes in the Czech Republic are phytotoxicity tests of elutriates, using S. alba and D. subspicatus, which have been found insufficiently sensitive to the range of different pollutants present in contaminated soils and/or solid wastes. If only aquatic bioassays are used for the toxicity testing, it is possible that the toxic effects of substances (poorly or totally) insoluble in water might be underestimated. The new proposed system of toxicity classification has proven to be both practical and sensitive. RECOMMENDATIONS AND PERSPECTIVES: This recommended alternative battery of phytotoxicity tests includes both aquatic tests of waste elutriates (with the algae D. subspicatus along with the aquatic plant L. minor), in addition to tests of the terrestrial solid samples (with the dicotyledonous L. sativa and the monocotyledonous H. vulgare). This battery of bioassays is sufficiently sensitive, representing a majority of types of aquatic and terrestrial plants. PMID- 19557448 TI - Phytoremediation of contaminated soils and groundwater: lessons from the field. AB - BACKGROUND, AIM, AND SCOPE: The use of plants and associated microorganisms to remove, contain, inactivate, or degrade harmful environmental contaminants (generally termed phytoremediation) and to revitalize contaminated sites is gaining more and more attention. In this review, prerequisites for a successful remediation will be discussed. The performance of phytoremediation as an environmental remediation technology indeed depends on several factors including the extent of soil contamination, the availability and accessibility of contaminants for rhizosphere microorganisms and uptake into roots (bioavailability), and the ability of the plant and its associated microorganisms to intercept, absorb, accumulate, and/or degrade the contaminants. The main aim is to provide an overview of existing field experience in Europe concerning the use of plants and their associated microorganisms whether or not combined with amendments for the revitalization or remediation of contaminated soils and undeep groundwater. Contaminations with trace elements (except radionuclides) and organics will be considered. Because remediation with transgenic organisms is largely untested in the field, this topic is not covered in this review. Brief attention will be paid to the economical aspects, use, and processing of the biomass. CONCLUSIONS AND PERSPECTIVES: It is clear that in spite of a growing public and commercial interest and the success of several pilot studies and field scale applications more fundamental research still is needed to better exploit the metabolic diversity of the plants themselves, but also to better understand the complex interactions between contaminants, soil, plant roots, and microorganisms (bacteria and mycorrhiza) in the rhizosphere. Further, more data are still needed to quantify the underlying economics, as a support for public acceptance and last but not least to convince policy makers and stakeholders (who are not very familiar with such techniques). PMID- 19557449 TI - Derivation of motor vehicle tailpipe particle emission factors suitable for modelling urban fleet emissions and air quality assessments. AB - BACKGROUND, AIM AND SCOPE: Urban motor vehicle fleets are a major source of particulate matter pollution, especially of ultrafine particles (diameters < 0.1 microm), and exposure to particulate matter has known serious health effects. A considerable body of literature is available on vehicle particle emission factors derived using a wide range of different measurement methods for different particle sizes, conducted in different parts of the world. Therefore, the choice as to which are the most suitable particle emission factors to use in transport modelling and health impact assessments presented as a very difficult task. The aim of this study was to derive a comprehensive set of tailpipe particle emission factors for different vehicle and road type combinations, covering the full size range of particles emitted, which are suitable for modelling urban fleet emissions. MATERIALS AND METHODS: A large body of data available in the international literature on particle emission factors for motor vehicles derived from measurement studies was compiled and subjected to advanced statistical analysis, to determine the most suitable emission factors to use in modelling urban fleet emissions. RESULTS: This analysis resulted in the development of five statistical models which explained 86%, 93%, 87%, 65% and 47% of the variation in published emission factors for particle number, particle volume, PM(1), PM(2.5) and PM(10), respectively. A sixth model for total particle mass was proposed but no significant explanatory variables were identified in the analysis. From the outputs of these statistical models, the most suitable particle emission factors were selected. This selection was based on examination of the statistical robustness of the statistical model outputs, including consideration of conservative average particle emission factors with the lowest standard errors, narrowest 95% confidence intervals and largest sample sizes and the explanatory model variables, which were vehicle type (all particle metrics), instrumentation (particle number and PM(2.5)), road type (PM(10)) and size range measured and speed limit on the road (particle volume). DISCUSSION: A multiplicity of factors need to be considered in determining emission factors that are suitable for modelling motor vehicle emissions, and this study derived a set of average emission factors suitable for quantifying motor vehicle tailpipe particle emissions in developed countries. CONCLUSIONS: The comprehensive set of tailpipe particle emission factors presented in this study for different vehicle and road type combinations enable the full size range of particles generated by fleets to be quantified, including ultrafine particles (measured in terms of particle number). These emission factors have particular application for regions which may have a lack of funding to undertake measurements, or insufficient measurement data upon which to derive emission factors for their region. RECOMMENDATIONS AND PERSPECTIVES: In urban areas motor vehicles continue to be a major source of particulate matter pollution and of ultrafine particles. It is critical that in order to manage this major pollution source methods are available to quantify the full size range of particles emitted for transport modelling and health impact assessments. PMID- 19557450 TI - Risk assessment of toxins derived from Bacillus thuringiensis-synergism, efficacy, and selectivity. AB - BACKGROUND, AIM, AND SCOPE: This review deals with publications concerning the mode of action of Bt proteins and their potential synergism with extrinsic factors. The aim was to assess the impact of those factors especially regarding selectivity and efficacy of Bt toxins and to discuss possible gaps in current risk assessment of genetically engineered plants expressing Bt toxins. MAIN FEATURES: The review shows that several extrinsic factors are able to influence the selectivity and efficacy of Bt toxins. The findings are seen as being relevant for risk assessment in Bt plants. This conclusion is derived by discussing current state of knowledge about the mode of action of Bt proteins, unexpected effects on non-target organism, and the way how modified Bt toxins are expressed in genetically engineered plants. RESULTS: Several publications have been identified that show that certain factors and synergism can impact efficacy and selectivity of Bt toxins. These extrinsic factors are various and include other Bt toxins or parts from the spore of Bacillus thuringiensis as well as certain enzymes, environmental stress, non-pathogenic microorganisms, and infectious diseases. DISCUSSION: Research on the underlying mechanism of observed synergism might help to explain some of the effects found in non-target organisms. In general, possible synergism of Bt toxins with extrinsic factors can be relevant for risk assessment of genetically engineered Bt plants since they expose a modified Bt toxin to the environment under various conditions and over a long period of time. CONCLUSIONS: Risk assessment of genetically engineered plants should put into question the general assumption of a high selectivity and a linear dose-response relationship in the toxicity of Bt proteins. Both selectivity and efficacy can be influenced by synergism, which can provoke unexpected and undesired effects in non-target organisms. PERSPECTIVES: It is suggested that systematic research be promoted on synergism between Bt toxins and potential extrinsic factors that could impact the spectrum of susceptible organisms. This research should become a prerequisite for risk assessment of Bt plants. PMID- 19557451 TI - Isolation and identification of new vasodilative substances in diesel exhaust particles. AB - BACKGROUND, AIM, AND SCOPE: We recently developed a new isolation method for diesel exhaust particles (DEP), involving successive extraction with H(2)O, sodium bicarbonate, and sodium hydroxide, in which the sodium hydroxide extract was found to consist of phenolic components. Analysis of the extract revealed that vasodilative-active nitrophenols are in DEP in significantly higher concentrations than those estimated by an earlier method involving a combination of solvent extraction and repeated chromatography. These findings indicated that our new procedure offers a simple, efficient, and reliable method for the isolation and identification of bioactive substances in DEP. This encouraged us to extend our work toward investigating new vasodilatory substances in the sodium bicarbonate extract. MATERIALS AND METHODS: DEP were collected from the exhaust of a 4JB1-type engine (ISUZU Automobile Co., Tokyo, Japan). GC-MS analysis was performed with a GCMS-QP2010 instrument (Shimadzu, Kyoto, Japan). RESULTS: DEP dissolved in 1-butanol was successively extracted with water, sodium bicarbonate, and then aqueous sodium hydroxide. The sodium bicarbonate extract was neutralized and the resulting mixture of acidic components was subjected to reverse-phase (RP) column chromatography followed by RP-HPLC with fractions assayed for vasodilative activity. This led to the identification of terephthalic acid, p hydroxybenzoic acid, isophthalic acid, phthalic acid, 3-hydroxy-4-nitrobenzoic acid, 4-hydroxy-3-nitrophenol, and 1,4,5-naphthalene tricarboxylic acid as components of DEP. DISCUSSION: The sodium bicarbonate extract was rich in aromatic carboxylic acid components. Repeated reverse-phase chromatography resulted in the successful isolation of several acidic substances including the new vasodilative materials, 4-hydroxy-3-nitrobenzoic acid, and 3-hydroxy-4 nitrobenzoic acid. CONCLUSIONS: Our new fractionation method for DEP has made possible the isolation of new vasodilative compounds from the sodium bicarbonate extract. PMID- 19557452 TI - Defining genes: a computational framework. AB - The precise elucidation of the gene concept has become the subject of intense discussion in light of results from several, large high-throughput surveys of transcriptomes and proteomes. In previous work, we proposed an approach for constructing gene concepts that combines genomic heritability with elements of function. Here, we introduce a definition of the gene within a computational framework of cellular interactions. The definition seeks to satisfy the practical requirements imposed by annotation, capture logical aspects of regulation, and encompass the evolutionary property of homology. PMID- 19557454 TI - [Patient safety and innovation]. PMID- 19557455 TI - [Regional differences in the prevalence of overweight and obesity at school entry in Germany : reality or artifact?]. AB - Discovering trends and regional differences in the prevalence of overweight and obesity in children and adolescents is an important task, bearing in mind the high public health relevance of this widespread health risk. It is all the more important to ensure a common methodological basis for the calculations underlying statements about trends and comparisons. Using the data of the German Health Examination Survey for Children and Adolescents (KiGGS), the potential effects of two methodological sources of error when calculating overweight and obesity prevalence at school entry are presented. Finally, simple recommendations for avoiding these errors are given. PMID- 19557453 TI - beta-Carotene conversion products and their effects on adipose tissue. AB - Recent epidemiological data suggest that beta-carotene may be protective against metabolic diseases in which adipose tissue plays a key role. Adipose tissue constitutes the major beta-carotene storage tissue and its functions have been shown to be modulated in response to beta-carotene breakdown products, especially retinal produced after cleavage by beta-carotene 15,15'-monooxygenase (BCMO1), and retinoic acid arising from oxidation of retinal. However, the possibility exists that beta-carotene in its intact form can also affect adipocyte function. Development of a knock out model and identification of a loss-of-function mutation have pointed out BCMO1 as being probably the sole enzyme responsible for provitamin A conversion into retinal in mammals. The utilisation of BCMO1(-/ )mice should provide insights on beta-carotene effect on its own in the future. In humans, intervention studies have highlighted the huge interindividual variation of beta-carotene conversion efficiency, possibly due to genetic polymorphisms, which might impact on response to beta-carotene. This brief review discusses the processes involved in beta-carotene conversion and the effect of cleavage products on body fat and adipose tissue function. PMID- 19557456 TI - [Aspects of the risks associated with vCJK from plasma derivatives from human plasma]. PMID- 19557457 TI - [Indoor air guideline values for saturated acyclic aliphatic C4-C11 aldehydes]. AB - To protect public health the German Working Group on Indoor Guidelines of the Federal Environmental Protection Agency and the States' Health Authorities is issuing indoor air guideline values. For health evaluation of indoor air n- and i alkanals with at least 4 C atoms no appropriate human data is available. Based on animal studies irritation has been identified as crucial adverse effect following inhalative exposure to C4 to C6 alkanals. However, sufficient data is available for C4 alkanals only. Inflammation of the nose epithelium followed by non neoplastic and degenerative lesions has been seen in inhalative subchronic studies with butanal and chronic studies with i-butanal. The lowest adverse effect level for continuous exposure is assessed by the Working Group as 16 mg butanal per cubic meter. By applying an interspecies factor of 1 and an intraspecies factor of 10 a so-called health hazard value of 2 mg butanal/m3 indoor air is obtained. Regarding the few and limited data on inhalation toxicity of the higher homologues concerning irritation a health hazard value of 2 mg/m3 indoor air for the total of acyclic C4 to C11 alkanals is recommended. Elevated C4 to C11 alkanal indoor air concentrations annoy by odour. To prevent from unpleasant odour a so-called health precaution value of 0.1 mg acyclic C4 to C11 alkanals/m3 indoor is recommended. PMID- 19557459 TI - [Multislice computed tomography in emergency radiology]. PMID- 19557460 TI - [Multidetector-row spiral computed tomography in chest emergencies]. AB - With ongoing advances in multidetector-row computed tomography (MDCT) using ECG gating, differentiated examination protocols have become technically feasible. For acute chest pain assessment a strict triage of patients is indispensable, as the radiation dose is approximately 3 times higher for a dedicated protocol compared to a standard chest MDCT. Clinical requests considering pathologies of the pulmonary arteries, the aortic arch and the descending aorta can safely be answered with a standard CT data set. However, for the coronary arteries as well as for the ascending aorta, ECG synchronization of the data set is required. Initial reports regarding MDCT assessment for acute chest pain report a high negative predictive value. With the latest MDCT platforms available, medical preparation is no longer necessary with the exception of sublingual application of nitroglycerine. Dedicated contrast injection protocols, however, are necessary for simultaneous opacification of the pulmonary arteries as well as of the aorta and the coronary arteries. Further prospective studies will have to provide more evidence-based data for acute chest pain assessment with MDCT and will also have to outline the cost-effectiveness of this imaging technique. PMID- 19557463 TI - [Urine-based oncological diagnostic for bladder carcinoma: what is new?]. PMID- 19557461 TI - [Multidetector computed tomography in abdominal emergencies]. AB - Abdominal emergencies encompass traumatic events in the region of the abdomen as well as the clinical term acute abdomen. Multidetector CT (MDCT) represents one of the most important imaging modalities for detection and correct diagnosis in such abdominal emergencies. Based on the acquired data MDCT allows a stable imaging of the abdomen in an extremely short time even in critically ill patients. Multiplanar reconstructions can be created based on the high resolution data allowing an impressive visualization of the pathology. In this review article the most important abdominal pathologies of the abdomen diagnosed by MDCT are presented with special emphasis on the liver, bile ducts, spleen and kidneys as well as the gastro-intestinal tract. Additionally, MDCT imaging of the individual organ regions will be dealt with in detail. PMID- 19557464 TI - [Urine-based tumour diagnostics for bladder cancer: effects of the new histopathological classification--food for thought]. AB - The new WHO classification of bladder cancer was published in 2004 and consequently cannot be regarded as very recent. However, it is still timely since it picks up considerations affecting other schemes of tumour classification as well. Genetic results are included in the context of morphology, and at the same time a high inter- and intra-observer agreement is striven for as a matter of high quality patient care. The WHO classification of 2004 does not include cytological diagnosis. Thinking about and considering tumour tissue diagnosis, the style of cytological diagnoses is also affected. For tissue diagnoses, low- and high-grade tumours are differentiated from benign lesions including reactive changes. The element of this classification which has to be transferred to cytology is especially the unequivocal diagnosis of high-grade lesions. The low grade lesion, correlating with tissue of well-differentiated papillary tumours and dysplasias, mostly cannot be distinguished cytologically with certainty from a broad spectrum of non-malignant lesions (papillomas, reactive urothelial detachment in urolithiasis patients, cytology specimen from vigorously irrigated bladders). For the latter group our aim should be to establish an additional diagnostic tool of high quality driven by clinical questions (e.g. potential of tumour progression). PMID- 19557465 TI - [Urine-based markers of angiogenesis in bladder cancer]. AB - Angiogenesis is a prerequisite for tumour growth and metastasis. Therefore, angiogenesis factors in bladder cancer, a common disease of the genitourinary tract, could serve as diagnostic tools, predictors of prognosis, and targets for therapy. Development of less invasive or noninvasive detection techniques, reliable prognostic markers, and individualized targeted therapy would have a significant impact on disease management. For this investigative goal, the utility of urine and blood is beneficial. Research in the field of angiogenesis and promising markers is currently evolving. In spite of the recent success of antiangiogenic agents in the oncological clinic, an optimal marker that will warrant substitution of the cystoscopic follow-up protocol in patients with urothelial neoplasms has not been identified yet. Despite this challenge, allocating more resources and attention to identifying such urine markers is justified to optimize the diagnostics and follow-up of urinary bladder cancer. PMID- 19557466 TI - [Urine-based cytological diagnosis against the background of the new histopathological classification]. AB - Urinary cytology is a basic adjunct to cystoscopy and transurethral resection in the diagnosis and characterization of high-grade urothelial carcinomas of the bladder. According to the new WHO classification the former tumor grading G1-3 for non-invasive carcinomas has been replaced by a separation into low-grade and high-grade urothelial carcinomas. An interesting question is where the former non invasive G2 carcinomas will be positioned in this new classification. In a retrospective analysis we focused on 44 patients with pTaG2 and 17 patients with pT1G2 carcinomas and found that this group of tumors is cytologically heterogeneous but easily differentiated into low-grade and high-grade lesions. A cytometrical analysis significantly underlines the results of the cytological diagnostics. High-grade tumors show a higher recurrence and progression rate. Cytological diagnostics can therefore assist in differentiating low-grade from high-grade urothelial carcinomas. PMID- 19557467 TI - [Urine-based diagnostics: an update on the Kiel Tumor Bank]. AB - The heterogeneity of bladder tumors in their ability to invade and metastasize and their frequent recurrence pose a challenge for physicians who treat bladder cancer patients and for the researchers who work on bladder cancer diagnosis, recurrence, and treatment-related areas. For most new bladder cancer cases, investigation begins when patients are symptomatic (i.e., hematuria or irritative voiding). This mode of detection is often inadequate for nearly 15-30% of these new cases with high-grade bladder cancer, since the tumor is already in the invasive stage at the time of diagnosis. Bladder cancer patients are on a mandatory 3-month to 6-month surveillance schedule because bladder tumors frequently recur. The current mode of detecting bladder cancer involves cystoscopy, which is an invasive and relatively expensive procedure. Voided urine cytology, the standard noninvasive marker, is highly tumor specific and has good sensitivity for detecting high-grade tumors. However, its sensitivity for detecting low-grade tumors is low; its accuracy depends on the examiner's expertise; and it is not available everywhere. Marker systems are readily available for use in practice. Their utility remains under discussion. PMID- 19557469 TI - [Systemic chemotherapy for bladder cancer: news in 2009]. AB - This review summarizes the results of first-line and second-line systemic chemotherapy for advanced urothelial carcinoma as well as data from adjuvant and neoadjuvant trials for locally advanced bladder cancer. Whereas conventional systemic chemotherapy prevailed for over two decades, targeted therapeutics have been introduced to treat urothelial cancer during recent years. As in other tumor entities, molecular profiling will presumably emerge in the future as a means to tailor individual therapy. PMID- 19557470 TI - [Premature ejaculation]. AB - With prevalence rates of 20%-25% premature ejaculation (PE) represents the most frequent sexual dysfunction in men. Whereas genetically determined changes in the serotonin receptor-/transporter mechanism seem to be responsible for lifelong PE, acquired PE is often associated with other conditioning diseases such as erectile dysfunction, prostatitis or thyroid dysfunctions. Typical features of PE are a short intravaginal ejaculatory latency time (IELT) <1-2 min, lack of control over ejaculation, personal distress and partner problems. Treatment of PE subdivides into sexual therapy as well as drug therapy. Among the medications considered for PE, oral therapy with selective serotonin re-uptake inhibitors (SSRI), Dapoxetine (the first officially approved medication for PE) and topical therapy with lidocaine/prilocaine-containing medications are given priority. PMID- 19557474 TI - Investigation of the genotoxicity of malathion to freshwater teleost fish Channa punctatus (Bloch) using the micronucleus test and comet assay. AB - Malathion [S-(1,2-dicarboethoxyethyl) O, O-dimethyl phosphorodithioate] is a widely used organophosphorus insecticide throughout the world. However, limited efforts have made to study its genotoxic effect in different fish tissues. The present investigation was aimed to assess the genotoxic potential of the pesticide to the freshwater teleost fish Channa punctatus at sublethal concentrations using the micronucleus test and comet assay. Initially, the 96-h LC50 value of commercial-grade malathion (50% EC) was determined as 5.93 ppm in a semistatic system. Based on LC50, three test concentrations (viz. sublethal I, sublethal II, and sublethal III) were determined to be 1.48, 0.74, and 0.59 ppm, respectively, and the fish specimens were exposed to these concentrations. Tissue samplings were done on days 0, 1, 3, 7, 15, 22 and 29 of malathion exposure for assessment of the induction of micronuclei (MN) frequency and DNA damage. The MN formation in the peripheral blood cells was found to be significantly higher (p < 0.05) in the treated specimens at all sampling intervals compared to the control. The MN frequency reached maximum on days 3 and 7 at sublethal I and II concentrations, respectively, followed by a nonlinear decline with the progression of the experiment. Similarly, significant effects (p < 0.05) of both concentration and time of exposure were observed on DNA damage in the gill, kidney, and lymphocytes. All of the tissues exhibited a concentration-dependent increase in DNA damage up to day 3, followed by a nonlinear decrease with the duration of exposure. A comparison of the extent of DNA damage among the tissues showed the sensitivity of gill tissue to malathion. PMID- 19557476 TI - Gadolinium use in spine pain management procedures for patients with contrast allergies: results in 527 procedures. PMID- 19557475 TI - Primary leiomyosarcoma of the inferior vena cava: case report. AB - The patient was a 63-year-old woman with a chief complaint of blood-stained sputum. A tumor of the inferior vena cava was found on chest computed tomography (CT) and identified as a primary tumor based on multidetector CT and contrast enhanced MR angiography. An intrapelvic tumor was also discovered. On autopsy, the two tumors were diagnosed as leiomyosarcoma and ovarian fibroma, respectively. PMID- 19557477 TI - A novel stable isotope approach for determining the impact of thickening agents on water absorption. AB - Research on the bioavailability of water from thickened fluids has recently been published and it concluded that the addition of certain thickening agents (namely, modified maize starch, guar gum, and xanthan gum) does not significantly alter the absorption of water from the healthy, mature human gut. Using xanthan gum as an example, our "proof of concept" study describes a simple, accurate, and noninvasive alternative to the methodology used in that first study, and involves the measurement and comparison of the dilution space ratios of the isotopes (2)H and (18)O and subsequent calculation of total body water. Our method involves the ingestion of a thickening agent labeled with (2)H 1 day after ingestion of (18)O. Analyses are based on the isotopic enrichment of urine samples collected prior to the administration of each isotope, and daily urine samples collected for 15 days postdosing. We urge that further research is needed to evaluate the impact of various thickening agents on the bioavailability of water from the developing gut and in cases of gut pathology and recommend our methodology. PMID- 19557478 TI - Complete soft tissue sarcoma resection is a viable treatment option for select elderly patients. AB - BACKGROUND: Decreased performance status, comorbidities, and disease natural history may erode enthusiasm for soft tissue sarcoma (STS) resection in elderly patients. Consequently, we evaluated the outcome of elderly patients amenable to complete surgical resection treated at a single institution. METHODS: Prospectively accrued data were used to identify patients with primary STS age >or=65 years (n = 325) who underwent complete macroscopic resection at our institution (1996-2007). Univariable and multivariable analyses were performed to identify prognostic factors. RESULTS: Median age at presentation was 72 years; 179 patients (55.1%) had associated comorbidities with an ASA score of >or=3. Extremity was the most common site (57.1%; n = 186), undifferentiated pleomorphic sarcoma the most common histology (60.4%; n = 197); 232 (71.2%) were high grade, 222 (68.3%) were >5 cm. Thirty-day postoperative mortality was 0.9% (n = 3); overall complication rate was 30.7% (n = 100), and mean postoperative hospital stay was 9 days (range, 1-84). Estimated median survival was 96 months, 5-year disease-specific survival (DSS) was 63%. Multivariable analysis identified age >or=75 year (HR = 2.03), tumor size: 5-15 vs <5 cm (HR = 3.54), or >15 vs <5 cm (HR = 10.33), and high-grade (HR = 5.53) as significant independent adverse prognostic factors. Compared with patients aged 65-74 years, older patients had more high grade tumors (P = .04), received chemotherapy less often (P < .0001), developed different patterns of recurrence (P < .05), and exhibited a shorter median survival (70 months; P = .05). CONCLUSIONS: Properly selected elderly patients can safely undergo extensive STS resections. Until more effective therapies become available, surgery in the elderly is indicated and remains the best means for STS control. PMID- 19557479 TI - The impact of obesity on outcomes following pancreatectomy for malignancy. AB - BACKGROUND: Obesity has previously been shown to correlate with higher stage and decreased survival in pancreatic cancer. The aim of this study was to determine the impact of obesity on operative outcomes, recurrence, and overall survival. METHODS: A review of our 1345 patient prospective hepatopancreaticobiliary database was performed to identify patients undergoing pancreatic resection from January 1991 to August 2008 for adenocarcinoma. Obesity was defined as a body mass index (BMI) > 30 kg/m(2). Data was analyzed using Wilcoxon, t test, and chi square methods. Survival was analyzed using log-rank analysis. Postoperative complications were assessed using a 5-point scale. P < .05 was considered significant. RESULTS: Of 306 patients undergoing pancreatic resection for pancreatic adenocarcinoma examined, 68 were defined as obese. There was no significant difference seen in length of stay, operative time, tumor size, or node status. Obese patients had a higher operative blood loss (median 650 vs. 400 mL, P = .0008). Obese patients were more likely to suffer postoperative complications (67.6% vs. 50.4%, P = .01). There was no significant difference seen in disease-free survival or overall survival (22.1 months for obese vs. 25.6 months for nonobese, P = .5; 19.8 months for obese vs. 23.5 months for nonobese, P = .46). CONCLUSION: Obese patients had a higher rate and greater severity of postoperative complications, with increased operative blood loss. However, obese patients did not demonstrate any significant difference in specific oncologic factors or survival. These data suggest an equivalent biologic effect of obesity on pancreatic cancer survival. PMID- 19557480 TI - Evaluation of the radiocobalt-labeled [MMA-DOTA-Cys61]-Z HER2:2395(-Cys) affibody molecule for targeting of HER2-expressing tumors. AB - PURPOSE: Imaging using positron emission tomography (PET) in the field of nuclear medicine is becoming increasingly important. The aim of this study was to develop a method for labeling of affibody molecules with radiocobalt for PET applications. PROCEDURES: The human epidermal growth factor receptors type 2 (HER2) binding affibody molecule DOTA-Z(2395)-C was radiolabeled with (57)Co (used as a surrogate of (55)Co). The binding specificity and cellular processing of the labeled compound was studied in vitro followed by in vivo characterization in normal and tumor-bearing mice. Furthermore, a comparative biodistribution study was performed with a (111)In-labeled counterpart. RESULTS: DOTA-Z(2395)-C was successfully labeled with radiocobalt with nearly quantitative yield. The compound displayed good retention on cells over time and high tumor accumulation of radioactivity in animal studies. Imaging studies showed clear visualization of HER2-positive tumors. Furthermore, the radiocobalt label provided better tumor-to organ ratios than (111)In. CONCLUSIONS: Radiocobalt is a promising label for affibody molecules for future PET applications. PMID- 19557481 TI - Measuring residents' perceived preparedness and skillfulness to deliver cross cultural care. AB - BACKGROUND: As patient populations become increasingly diverse, we need to be able to measure residents' preparedness and skillfulness to provide cross cultural care. OBJECTIVE: To develop a measure that assesses residents' perceived readiness and abilities to provide cross-cultural care. DESIGN: Survey items were developed based on an extensive literature review, interviews with experts, and seven focus groups and ten individual interviews, as part of a larger national mailed survey effort of graduating residents in seven specialties. Reliability and weighted principal components analyses were performed with items that assessed perceived preparedness and skillfulness to provide cross-cultural care. Construct validity was assessed. PARTICIPANTS: A total of 2,047 of 3,435 eligible residents participated (response rate = 60%). MEASUREMENTS AND MAIN RESULTS: The final scale consisted of 18 items and 3 components (general cross-cultural preparedness, general cross-cultural skillfulness, and cross-cultural language preparedness and skillfulness), and yielded a Cronbach's alpha = 0.92. Construct validity was supported; the scale total was inversely correlated with a measure of helplessness when providing care to patients of a different culture (p < 0.001). CONCLUSIONS: We developed a three-component cross-cultural preparedness and skillfulness scale that was internally consistent and demonstrated construct validity. This measure can be used to evaluate residents' perceived effectiveness of cross-cultural medical training programs and could be used in future work to validate residents' self assessments with objective assessments. PMID- 19557482 TI - Aberrant insertion of the right subclavian artery: an unusual cause of dysphagia in an adult. AB - INTRODUCTION: Within this report, we present a patient with difficulty of swallowing caused by an aberrant right subclavian artery. DISCUSSION: This is a congenital anomaly with the right subclavian artery originating from the dorsal part of the aortic arch and coursing through the mediastinum between the esophagus and the vertebral column. The diagnosis and treatment of this disorder is discussed based on the findings from the literature. PMID- 19557483 TI - Is blind pancreaticoduodenectomy justified for patients with ampullary neoplasms? AB - BACKGROUND: Many specialists justify pancreaticoduodenectomy (PD) for pancreatic head neoplasms with suspected but unproven malignance (blind-PD). Our aim in this study was to determine whether blind-PD is also justified for ampullary neoplasms. METHODS: We retrospectively reviewed the records of all patients with presumed resectable ampullary neoplasms treated at the National Taiwan University Hospital from 1998 to 2008. RESULTS: Of the 84 patients without a preoperative tissue diagnosis of malignance, 64 had blind-PD and 20 had ampullectomy (AMP) with intraoperative frozen section. Patients with jaundice, gastrointestinal bleeding, imaging findings showing tumor invasion, and larger tumor size were significantly more frequently treated by blind-PD. Final pathological diagnosis was benign in ten of 64 blind-PD-treated patients. CONCLUSIONS: Our data support a selective use of blind-PD because (1) a significant portion (65%) of benign ampullary neoplasms can be safely and effectively treated by AMP, (2) blind-PD does not treat ampullary cancer at earlier stage, and (3) blind-PD is associated with significantly more complications and significantly longer hospital stay than AMP. However, blind-PD is strongly recommended for patients with large ampullary neoplasms (>3 cm in diameter), with jaundice, or with malignant endoscopic appearance. PMID- 19557484 TI - Risk of gastrojejunal anastomotic stricture with multifilament and monofilament sutures after hand-sewn laparoscopic gastric bypass: a prospective cohort study. AB - BACKGROUND: Gastrojejunal (GJ) stricture is one of the most common late complications after laparoscopic Roux-en-Y gastric bypass (LRYGBP) with a hand sewn anastomosis. The object of this study was to assess the risk of stricture for two types of resorbable suture (multifilament and monofilament) in a series of LRYGBPs performed by the same surgeon. METHODS: DESIGN: Prospective cohort study. The study population consisted of a series of consecutive morbidly obese patients who underwent primary hand-sewn LRYGBP between March 2004 and May 2008 at the University Hospital in Getafe, Madrid, Spain. The study comprised 242 LRYGBPs with a four-layer continuous hand-sewn anastomosis using absorbable 3/0 gauge suture. The suture material was Ethicon Vicryl multifilament in the first 105 cases and Ethicon Monocryl monofilament in the following 137 cases. All patients were followed up monthly for the first 6 months and then every 6 months after that. RESULTS: The mean BMI was 46 +/- 4 for the multifilament cohort and 48 +/- 6 for the monofilament cohort with no significant difference between the two (p = 0.567). There were no anastomotic leaks, and no cases of marginal ulcer, abscess, abdominal sepsis, deep vein thrombosis, or pulmonary embolism were recorded. No cases required conversion to open surgery, and perioperative mortality was zero. In all, 11 cases of stricture (4.4%) were recorded, 10 in the multifilament suture cohort (9.5%), and only one in the monofilament suture cohort (0.7%; p = 0.001). The odds ratio was 14.3 (95% CI = 1.8-113.4). The mean outpatient follow-up period was 30 months (range = 6-42). CONCLUSIONS: Anastomotic GJ stricture is a common and well-known complication of laparoscopic gastric bypass for morbid obesity. Hand sewing with monofilament suture significantly lowered the frequency of this complication, and hence, monofilament should be the suture material of choice for this suturing technique. PMID- 19557486 TI - Fenestrated cannulae with outflow reduces fluid gain in shoulder arthroscopy. AB - Soft tissue fluid retention is a common problem after arthroscopy, with as much as 2% of patients having complications develop. A fenestrated outflow cannula has been introduced to reduce interstitial swelling. We tested the ability of this outflow cannula design to reduce fluid weight gain. We enrolled 28 patients undergoing shoulder arthroscopy and randomized them into two groups using fenestrated outflow versus conventional cannulae. The conventional group had greater weight gain as a function of the procedure duration than the fenestrated outflow group (slope = 0.542 +/- 1.160 kg/hour versus 0.0144 +/- 0.932 kg/hour). The conventional group also had greater weight gain as a function of fluid volume than the fenestrated outflow group (slope = 0.022 +/- 0.038 kg/L versus 0.002 +/- 0.341 kg/L). Compared with conventional nonoutflow cannulae, fenestrated outflow cannulae with negative pressure reduced weight gain associated with longer arthroscopic surgeries and increased arthroscopic fluid volume. LEVEL OF EVIDENCE: Level I, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19557485 TI - Plasma glucose and insulin regulation is abnormal following gastric bypass surgery with or without neuroglycopenia. AB - BACKGROUND: Enhanced insulin sensitivity is commonly seen following Roux-en-Y gastric bypass surgery (RYGB) whereas symptomatic hypoglycemia post-RYGB seems to occur infrequently. It is unclear how different plasma glucose and insulin responses are in patients with symptomatic hypoglycemia (SX-RYGB) versus those who remain asymptomatic (ASX-RYGB), nor when compared with non-surgical controls with varying degrees of insulin sensitivity. METHODS: Plasma glucose and insulin concentrations were determined following a 75-g oral glucose challenge in five groups: symptomatic and asymptomatic patients following RYGB (n = 9 each) and overweight/obese controls, divided into three subgroups (n = 30 each) on the basis of degree of insulin sensitivity measured by the insulin suppression test. RESULTS: SX-RYGB group had higher 30-min glucose after oral glucose compared with the ASX-RYGB group (p = 0.04). The two groups did not differ in peak glucose and insulin concentrations, nadir glucose concentration, or insulin-to-glucose ratio 30 min after oral glucose. These values were significantly different from the three control groups, and peak insulin concentrations post-RYGB were increased at every degree of insulin sensitivity as compared with the control groups. CONCLUSIONS: Plasma glucose and insulin responses to oral glucose in patients with symptomatic hypoglycemia post-RYGB are minimally different when compared to individuals who remain asymptomatic, and both groups demonstrate hyperinsulinemia out of proportion to their degree of insulin sensitivity. PMID- 19557488 TI - Fine wigns. AB - There are three categories of diagnostic data collected during the physical examination-sign, symptom, and that in between-but only two words to describe them. To fill that gap, I modestly propose here that examination responses that are vocalized (and thus symptomlike) yet thought to be specific (hence signifiers) be classified as wigns. I define a wign as the subjective reaction to a provocative examination maneuver deemed to have some valid relationship to the underlying pathology. This word is pronounced "whine" to remind us it is a spoken response, and its spelling echoes that of sign, reminding us likewise a wign is more definitive than a generalized complaint. The distinctions between sign, symptom, and wign are worthy of preservation, particularly regarding their probative value: treatments offered on the basis of signs can be said to be most rigorously indicated, as symptoms, unlike signs, pass through (and are affected by) the prism of patients' perceptions. Remaining skeptical about the value of information provided by our patients is in the interest of these patients, as our skepticism might save them from unnecessary treatments and procedures. PMID- 19557487 TI - Starch-poly-epsilon-caprolactone microparticles reduce the needed amount of BMP 2. AB - BMP-2 is currently administered clinically using collagen matrices often requiring large amounts of BMP-2 due to burst release over a short period of time. We developed and tested a novel injectable drug delivery system consisting of starch-poly-epsilon-caprolactone microparticles for inducing osteogenesis and requiring smaller amounts of BMP-2. We evaluated BMP-2 encapsulation efficiency and the in vitro release profile by enzyme-linked immunosorbent assay. BMP-2 was rapidly released during the first 12 hours, followed by sustained release for up to 10 days. We then evaluated the osteogenic potential of dexamethasone (standard osteogenic induction agent) and BMP-2 after incorporation and during release using an osteo/myoblast cell line (C2C12). Alkaline phosphatase activity was increased by released BMP-2. Mineralization occurred after stimulation with BMP-2 loaded microparticles. A luciferase assay for osteocalcin promoter activity showed high levels of activity upon treatment with BMP-2-loaded microparticles. In contrast, no osteogenesis occurred in C2C12 cells using dexamethasone-loaded microparticles. However, human adipose stem cells exposed to the microparticles produced high amounts of alkaline phosphatase. The data suggest starch-poly epsilon-caprolactone microparticles are suitable carriers for the incorporation and controlled release of glucocorticoids and growth factors. Specifically, they reduce the amount of BMP-2 needed and allow more sustained osteogenic effects. PMID- 19557489 TI - Survivorship of a low-stiffness extensively porous-coated femoral stem at 10 years. AB - A novel low-stiffness extensively porous-coated total hip femoral component was designed to achieve stable skeletal fixation, structural durability, and reduced periprosthetic femoral stress shielding. In short- to intermediate-term clinical review, this implant achieved secure biologic fixation and preserved periprosthetic bone. We retrospectively reviewed all 102 prospectively followed patients (106 implants) with this implant to document the longer-term implant survivorship, clinical function, fixation quality, and periprosthetic bone preservation. Ninety-seven patients with 101 implants had current followup or were followed to patient death (range, 1-14 years; average, 10 years). Eighty-six living patients were followed for an average implant survivorship of 10 years. There were no known femoral implant removals. The average Harris hip score at 10 year followup was 98. Radiographs demonstrated secure implant fixation and maintenance of periprosthetic bone. These data suggest this implant design provided long-term function characterized by extensive fixation, structural durability, and radiographic appearance of maintained periprosthetic cortical thickness and density. LEVEL OF EVIDENCE: Level I, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19557490 TI - Predictors of success on the American Board of Orthopaedic Surgery examination. AB - Predictors of success of orthopaedic residents on the American Board of Orthopaedic Surgery (ABOS) examination are controversial. We therefore evaluated numerous variables that may suggest or predict candidate performance on the ABOS examination. We reviewed files of 161 residents (all graduates) from one residency program distributed into two study groups based on whether they passed or failed their first attempt on the ABOS Part I or Part II examination from 1991 through 2005. Predictors of success/failure on the ABOS I included the mean percentile score on the Orthopaedic In-Training Examination (OITE) (Years 2 through 4), the percentile OITE score in the last year of training, US Medical Licensing Examination (USMLE) score, Dean's letter, election to Alpha Omega Alpha (AOA), and number of honors in selected third-year clerkships. All but the USMLE score predicted passing the ABOS Part II examination. These data suggest there are objective predictors of residents' performance on the ABOS Part I and Part II examinations. PMID- 19557491 TI - End-result study of vitallium cup arthroplasties of the hip Harry E. Emmel MD, John F. LeCocq MD, Edward A. LeCocq MD, Darrell G. Leavitt MD, Harry L. Leavitt MD, Kirk J. Anderson MD, Henry H. Nash MD, J. Garth Mooney MD CORR 1958;11:41-50. PMID- 19557494 TI - Does inadequate diet during childhood explain the higher high fracture rates in the Southern United States? AB - SUMMARY: Southern states have the highest age-adjusted hip fracture rates among older adults in the United States. Regional hip fracture rates in the United States in 1986-1993 correlate with death rates from rickets in the 1940s. Historical patterns of bone nutrition early in life might explain contemporary geographic patterns in bone fragility. INTRODUCTION: State of residence early in life is a better predictor of the risk of hip fracture after age 65 than state of current residence. Therefore, the geography of rickets mortality in the United States before 1950 was compared with the geography of hip fracture rates among older adults in the United States during 1986-1993. METHODS: Vital statistics data for the US white population for 1942-1948 allowed calculation of the ratio of deaths from rickets to live births for each geographic division of the USA. These ratios were correlated with previously published, standardized hip fracture rates among whites 65-89 years old during 1986-1993 by census division. RESULTS: During 1942-1948, the rickets mortality ratio among whites was 3.11 in the South, 1.91 in the Northeast, 1.75 in the Midwest, and 1.04 in the West. The correlation of mortality with risk of hip fracture was 0.71 (p = 0.03) for both sexes combined and 0.86 (p = 0.01) for women. CONCLUSIONS: Inadequate nutrition during skeletal formation early in life might explain the higher incidence of hip fracture among older adults in the South. PMID- 19557495 TI - Internal carotid artery dissection and stroke after SCUBA diving: a case report and review of the literature. PMID- 19557496 TI - Obsessive-compulsive traits in children and adolescents with Asperger syndrome. AB - The objective of this study is to examine the occurrence and characteristic features of obsessive-compulsive behaviours in children and adolescents with Asperger syndrome (AS), with respect to a matched obsessive compulsive disorder group (OCD) and a typically developing control group (CG). For this purpose, 60 subjects (20 OCD; 18 AS; 22 CG), aged 8-15 years, matched for age, gender and IQ were compared. AS and OCD patients were diagnosed according to the DSM-IV-TR criteria. The Autism Diagnostic Interview-Revised and the Autism Diagnostic Observation Schedule were used to assist in the AS diagnosis; the WISC-R was administered to assess IQ. Obsessive and compulsive symptoms were evaluated by using the Children's Yale-Brown Obsessive-Compulsive Scale (CY-BOCS). None of the AS children received a formal diagnosis of OCD. The AS group presented significantly higher frequencies of Hoarding obsessions and Repeating, Ordering and Hoarding compulsions compared to CG. The OCD group, in turn, reported significantly higher frequencies of Contamination and Aggressive obsessions and Checking compulsions compared to both the AS group and CG. As expected, the OCD group displayed a higher severity of symptoms (Moderate level of severity) than did the AS group (Mild level of severity). Finally, in our sample, neither the OCD group nor the AS group demonstrated a completely full awareness of the intrusive, unreasonable and distressing nature of symptoms, and the level of insight did not differ between the OCD group and CG, although an absence of insight was observed in the AS group. Children with AS showed higher frequencies of obsessive and compulsive symptoms than did typically developing children, and these features seem to cluster around Hoarding behaviours. Additionally, different patterns of symptoms emerged between the OCD and AS groups. Finally, in our sample, the level of insight was poor in both the OCD and the AS children. Further research should be conducted to better understand the characteristics of repetitive thoughts and behaviours in autism spectrum disorders, and to clarify the underlying neurobiological basis of these symptoms. PMID- 19557497 TI - Surface display of IgG Fc on baculovirus vectors enhances binding to antigen presenting cells and cell lines expressing Fc receptors. AB - Recombinant baculoviruses (recBV) were constructed with dual cassettes for constitutive expression of human IgG Fc following infection of insect cells and the structural proteins of hepatitis C virus (core, E1 and E2) following transduction of mammalian cells. The IgG Fc was expressed in insect cells as a fusion protein with the signal sequence and transmembrane region of either the native baculovirus envelope protein gp64 or the human transferrin receptor as a type I or type II integral membrane protein, respectively. The IgG Fc fusion proteins formed functional homodimers on the surface of recBV-infected insect cells and were incorporated into the envelope of recBV particles during egress from the infected cell. Both pseudotyped recBV bound specifically to recombinant soluble FcgammaRIIalpha receptor and to cell lines and antigen-presenting cells expressing Fc receptors (FcRs). These novel baculoviral vectors, which target cells of the immune system that express FcRs, have potential applications for vaccination or gene therapy. PMID- 19557498 TI - Oryzalin bodies: in addition to its anti-microtubule properties, the dinitroaniline herbicide oryzalin causes nodulation of the endoplasmic reticulum. AB - Oryzalin is a much-used pre-emergence herbicide which causes microtubules (Mt) to depolymerize. Here, we document that this dinitroaniline herbicide also leads to characteristic changes in the morphology of the endoplasmic reticulum (ER) and Golgi apparatus. These effects, which are reversible upon washing out the herbicide, are already elicited at low concentrations (2 microM) and become most pronounced at 20 microM. For our studies, we have employed roots of Arabidopsis thaliana, tobacco leaf epidermal cells, and BY-2 suspension cultures, all expressing the luminal ER marker GFP::HDEL. In all cell types, the typical cortical network of the ER assumed a pronounced nodulated morphology with increasing oryzalin concentrations. This effect was enhanced through subsequent application of brefeldin A (BFA). Thin sections of Arabidopsis roots observed in the electron microscope revealed the nodules to consist of a mass of anastomosing ER tubules. Oryzalin also caused the cisternae in Golgi stacks to increase in number but reduced their diameter. Oryzalin retarded ER mobility but did not prevent latrunculin B-induced clustering of Golgi stacks on islands of cisternal ER. While the mechanism underlying these changes in endomembranes remains unknown, it is specific for oryzalin since these effects were not elicited with other Mt-depolymerizing herbicides, e.g., trifluralin, amiprophosmethyl, or colchicine. PMID- 19557499 TI - Neurological outcome of long-term glioblastoma survivors. AB - Extended survival of 3 or more years is rare in patients with glioblastoma (GBM) but is becoming more common. Clinical outcome has not been well studied. We reviewed GBM patients at Memorial Sloan-Kettering Cancer Center between 2001 and 2003 who were seen for two or more visits. Patient characteristics and long-term clinical outcomes were reviewed for patients who had survived 3 or more years following diagnosis. Thirty-nine (11%) of 352 GBM patients were identified as long-term survivors. Median survival was 9.15 years (range: 3-18 years). Median age was 47 years (range: 16-69); 13% were 65 years or older. Median KPS was 90 (range: 50-100). One long-term survivor underwent biopsy alone; 19 patients each had either complete or subtotal resection. All received focal radiotherapy (RT) with a median dose of 5940 cGy; 18% received concurrent temozolomide. Adjuvant chemotherapy was administered to 35 (90%). Twelve patients (31%) remained in continuous remission. Twenty-seven had tumor progression a median of 29.2 months after diagnosis (range: 1.2-167 months); 18 had multiple relapses. Median KPS at last follow-up was 70 (range: 40-100); 85% of long-term survivors had at least one significant neurologic deficit. Eleven (28%) had clinically significant RT induced leukoencephalopathy, 9 (23%) developed RT necrosis and 9 (23%) treatment related strokes. Treatment-related complications occurred a median of 2.7 years from diagnosis (range: 0.9-11.5 years). Long-term survivors remain rare, but are found across all age groups despite multiple recurrences; clinically significant delayed complications of treatment are common. PMID- 19557500 TI - Evaluation of neo-angiogenesis in a case of chordoid meningioma. AB - Chordoid is a rare variant of meningioma that displays a high recurrence rate. In this study we evaluated the immunohistochemical expression of factors that regulate the neo-angiogenesis process in a case of chordoid meningioma with an adverse clinical course. A high microvessel density (MVD), as well as high VEGF and VEGF-R1 expression, was encountered in this tumor. By contrast, the anti angiogenic factor semaphorin3A, highly expressed in this chordoid meningioma, was ineffective because of the absence of its receptor neuropilin-1. We suggest that VEGF expression and MVD may represent significant prognostic factors in chordoid meningiomas. PMID- 19557501 TI - Clinical improvement and survival in breast cancer leptomeningeal metastasis correlate with the cytologic response to intrathecal chemotherapy. AB - Leptomeningeal meningitis occurs in approximately 5% of metastatic breast cancers, and there is no standard treatment for this complication. We retrospectively analyzed the clinical data and cerebrospinal fluid of 24 patients treated with high-dose intrathecal methotrexate for breast cancer leptomeningeal meningitis (BLM). Cytologic response (CSF cytology without neoplastic cells after treatment) was observed in 11 patients (46%) and related to survival (P = 0.005). In addition, clinical symptoms improved in all 11 patients who had a cytologic response and in 7 patients (54%) without cytologic response (P = 0.02). The predictive value of cytologic response needs further confirmation. Cytologic response could be helpful in the management of intrathecal chemotherapy in patients with BLM. PMID- 19557502 TI - RIOK3 interacts with caspase-10 and negatively regulates the NF-kappaB signaling pathway. AB - RIOK3 was initially characterized as a homolog of Aspergillus nidulans sudD and showed down-regulation at the invasive front of malignant melanomas, but the molecular mechanism remains elusive. Here, we report that overexpression of RIOK3 inhibits TNFalpha-induced NF-kappaB activation, but down-regulation of endogenous RIOK3 expression by siRNA potentiates it. A yeast two-hybrid experiment revealed that RIOK3 interacted with caspase-10, and further, a GST pull-down assay and endogenous coimmunoprecipitation validated the interaction. We subsequently showed that the interaction was mediated by the RIO domain of RIOK3 and each death effector domain of caspase-10. Interestingly, our data demonstrated that RIOK3 suppressed caspase-10-mediated NF-kappaB activation by competing RIP1 and NIK to bind to caspase-10. Importantly, the kinase activity of RIOK3 was confirmed to be relevant to NF-kappaB signaling. Taken together, our findings strongly suggest that RIOK3 negatively regulates NF-kappaB signaling pathway activated by TNFalpha dependent on its kinase activity and NF-kappaB signaling pathway activated by caspase-10 independent of its kinase activity. PMID- 19557504 TI - Fabrication of a novel core-shell gene delivery system based on a brush-like polycation of alpha, beta-poly (L-aspartate-graft-PEI). AB - PURPOSE: A novel core-shell gene delivery system was fabricated in order to improve its gene transfection efficiency, particularly in the presence of serum. MATERIALS AND METHODS: alpha, beta-poly (L-aspartate-graft-PEI) (PAE) was simply synthesized by ring-opening reaction of poly (L-succinimide) with low molecular weight (LMW) linear polyethylenimine (PEI, Mn = 423). PAE/DNA nanoparticles were characterized. Condensation and protection ability of plasmid by PAE were confirmed by agarose gel electrophoresis assay. Cytotoxicity of the polymer and polymer/DNA nanoparticles were measured by MTS assay. Gene transfection efficiencies were evaluated both in vitro and in vivo. RESULTS: Core-shell nanoparticles assembled between DNA and PAE showed positive zeta potential, narrow size distribution, and spherical compact shapes with size below 250 nm when N/P ratio is above 10. Cytotoxicity of PAE was rather lower than that of PEI 25K, while the most efficient gene transfection and serum resistant ability of PAE/DNA complexes were higher than that of PEI 25K. Bafilomycin A1 treatment suggested "proton sponge" mechanism of PAE-mediated gene transfection. PAE/pEGFP N2 nanoparticles also showed good gene expression in vivo and were dominantly distributed in kidney, liver, spleen and lung after intravenous administration. CONCLUSIONS: The results demonstrated the potential use of PAE as an effective gene carrier. PMID- 19557503 TI - Egr-1 upregulates OPN through direct binding to its promoter and OPN upregulates Egr-1 via the ERK pathway. AB - Early growth response factor-1 (Egr-1) and osteopontin (OPN) play important roles in the migration and proliferation of vascular smooth muscle cells (VSMC), but little is known about their relationship. Therefore, we transfected VSMCs with either Egr-1 cDNA, Opn cDNA, a DNA enzyme designed to target Egr-1 (ED5), or antisense Opn oligodeoxynucleotides and examined changes in Egr-1 and OPN expression at the mRNA and protein levels. OPN expression levels were increased in cells that were stably transfected with Egr-1 cDNA. By contrast, both Egr-1 and OPN expression were reduced when ED5 was transfected into Egr-1-expressing cells. Similarly, Opn transfection upregulated Egr-1 levels, while Opn anti-sense oligodeoxynucleotide transfection decreased Egr-1 expression. ChIP analysis showed that Egr-1 binds to the Opn gene promoter. Furthermore, treatment with the extracellular-regulated protein kinase (ERK) inhibitor PD98059 inhibited the upregulation of Egr-1 by OPN. We find that Egr-1 and OPN positively regulate each other in VSMCs. PMID- 19557505 TI - Self-reported functional status as predictor of observed functional capacity in subjects with early osteoarthritis of the hip and knee: a diagnostic study in the CHECK cohort. AB - OBJECTIVES: Patients with hip or knee osteoarthritis (OA) may experience functional limitations in work settings. In the Cohort Hip and Cohort Knee study (CHECK) physical function was both self-reported and measured performance-based, using Functional Capacity Evaluation (FCE). Relations between self-reported scores on SF-36 and WOMAC (Western Ontario and McMaster Arthritis Index, function scales) and FCE performance were studied, and their diagnostic value for clinicians in predicting observed physical work limitations was assessed. METHODS: Ninety-two subjects scored physical function on SF-36 (scale 0-100, 100 indicating the best health level) and WOMAC (scale 0-68, 68 indicates maximum restriction) and performed the FCE. Correlations were calculated between all scores. Cross-tables were constructed using both questionnaires as diagnostic tests to identify work limitations. Subjects lifting <22.5 kg on the FCE-test 'lifting-low' were labeled as having physical work limitations. Diagnostic aspects at different cut-off scores for both questionnaires were analysed. RESULTS: Statistically significant correlations (Spearman's rho 0.34-0.49) were found between questionnaire scores and lifting and carrying tests. Results of a diagnostic cross-table with cut-off point <60 on SF-36 'physical functioning' were: sensitivity 0.34, specificity 0.97 and positive predictive value (PV+) 0.95. Cut-off point > or =21 on WOMAC 'function' resulted in sensitivity 0.51, specificity 0.88 and PV+ 0.88. CONCLUSION: Low self-reported function scores on SF-36 and WOMAC diagnosed subjects with limitations on the FCE. However, high scores did not guarantee performance without physical work limitations. These results are specific to the tested persons with early OA, in populations with a different prevalence of limitations, different diagnostic values will be found. FCE may be indicated to help clinicians to assess actual work capacity. PMID- 19557506 TI - Fluorescence interaction and determination of calf thymus DNA with two ethidium derivatives. AB - In this paper, we reported the syntheses and investigation of the modes of binding to DNA of the two new ethidium derivatives containing benzoyl and phenylacetyl groups of both amines at 3-and 8- positions. The interactions between calf thymus DNA (ct-DNA) and the two derivatives, 3,8-dibenzoylamino-5 ethyl-6-phenylphenantridinium cloride (E2) and 3,8-diphenylacetylamino-5-ethyl-6 phenylphenantridinium chloride (E3), were investigated by fluorescence quenching spectra and UV-vis absorption spectra. The Stern-Volmer quenching constants, binding constants, binding sites and the corresponding thermodynamic parameters DeltaH, DeltaS and DeltaG were calculated at different temperatures. The results indicated the formation of E2 and E3-DNA complexes and van der Waals interactions as the predominant intermolecular forces in stabilizing for each complex. In addition, increasing nucleophilicity of the functional groups at 3- and 8- positions exhibited the respectable increment the DNA binding affinities of derivatives. The results of absorption, ionic strength and iodide ion quenching suggested that the interaction mode of E2 and E3 with ct-DNA was intercalative binding. The limit of detection (LOD) of ct-DNA were 7.49 x 10(-8) (n = 4) and 4.18 x 10(-8) mol/l (n = 7) in presence of E2 and E3, respectively. PMID- 19557507 TI - Socioeconomic gradients in infant health across race and ethnicity. AB - The objective of this study was to examine socioeconomic (SES) gradients in infant health across a number of racial and ethnic groups in the United States. The study was based on data from a new nationally representative sample of children born in the US in 2001 (N = 8,650). The data include oversamples of several minority groups and a rich set of socioeconomic indicators, as well as demographic, health, and health behavior characteristics. Proportion of low birth weight (LBW) and small for gestational age (SGA) (and 95% CIs) across categories of several indicators of SES (maternal education, income, income adjusted for family size, and wealth) was presented for the full sample of children and disaggregated by race/ethnicity: non-Hispanic white, non-Hispanic black, Hispanic, Asian/Pacific Islander (A/PI), and American Indian/Alaskan Native (AI/AN). A graded relationship was found between all measures of SES and infant health for white mothers, and between adjusted income and LBW for Asian and Hispanic mothers. There was no relationship between any indicator of SES and either LBW or SGA for either black or AI/AN mothers. The finding that some minority racial/ethnic groups do not reap the same health benefits from higher levels of SES as do whites suggests that approaches to reducing health disparities must address not only the structural barriers that lead some minority groups to have fewer resources but also barriers that prevent these groups from taking advantage of those resources to improve the health of their families. PMID- 19557508 TI - Disparities in initiation and adherence to prenatal care: impact of insurance, race-ethnicity and nativity. AB - We used the intersectionality framework to examine impact of racial/ethnic, immigration, and insurance differences on the timing of initiation of prenatal care (PNC) and subsequent adherence. In this cross sectional study independent variables were women's race/ethnicity; nativity; age; education; and insurance. The dependent variables were late initiation and non-adherence to recommended number of PNC visits. We used multivariate analysis to evaluate the impact of the independent variables on late initiation and non-adherence. Analysis revealed that race/ethnicity/nativity (RE-N) was more consistently associated with late initiation and non-adherence for privately insured than publicly insured persons. While private insurance had a positive impact on initiation and adherence overall, its impact was greater for White women. Having private insurance coverage was most beneficial to White women. We contend that the intersectional approach provides promising avenues for expanding our knowledge of health disparities and of identifying new ways of going about eliminating the persistent and pervasive social inequalities and informing efforts to reduce them. PMID- 19557509 TI - Human versus non-human face processing: evidence from Williams syndrome. AB - Increased motivation towards social stimuli in Williams syndrome (WS) led us to hypothesize that a face's human status would have greater impact than face's orientation on WS' face processing abilities. Twenty-nine individuals with WS were asked to categorize facial emotion expressions in real, human cartoon and non-human cartoon faces presented upright and inverted. When compared to both chronological and mental age-matched controls, WS participants were able to categorize emotions from human, but not from non-human faces. The use of different perceptual strategies to process human and non-human faces could not explain this dissociation. Rather, the findings suggest an increased sensitivity to socially relevant cues, such as human facial features, possibly related to the hallmark feature of WS-hypersociability. PMID- 19557510 TI - Similarities between simulated spatial spectra of scalp EEG, MEG and structural MRI. AB - Electrical dipoles oriented perpendicular to the cortical surface are the primary source of the scalp EEGs and MEGs. Thus, in particular, gyri and sulci structures on the cortical surface have a definite possibility to influence the EEGs and MEGs. This was examined by comparing the spatial power spectral density (PSD) of the upper portion of the human cortex in MRI slices to that of simulated scalp EEGs and MEGs. The electrical activity was modeled with 2,650 dipolar sources oriented normal to the local cortical surface. The resulting scalp potentials were calculated with a finite element model of the head constructed from 51 segmented sagittal MR images. The PSD was computed after taking the fast Fourier transform of scalp potentials. The PSD of the cortical contour in each slice was also computed. The PSD was then averaged over all the slices. This was done for sagittal and coronal view both. The PSD of EEG and MEG showed two broad peaks, one from 0.05 to 0.22 cycles/cm (wavelength 20-4.545 cm) and the other from 0.22 to 1.2 cycles/cm (wavelength 4.545-0.834 cm). The PSD of the cortex showed a broad peak from 0.08 to 0.32 cycles/cm (wavelength 12.5-3.125 cm) and other two peaks within the range of 0.32 to 0.9 cycles/cm (wavelength 3.125-1.11 cm). These peaks are definitely due to the gyri structures and associated larger patterns on the cortical surface. Smaller peaks in the range of 1-3 cycles/cm were also observed which are possibly due to sulci structures. These results suggest that the spatial information was present in the EEG and MEG at the spatial frequencies of gyri. This also implies that the practical Nyquist frequency for sampling scalp EEGs should be 3.0 cycles/cm and an optimal interelectrode spacing of about 3 mm is needed for extraction of cortical patterns from scalp EEGs in humans. PMID- 19557512 TI - Preface: Role of cell surface macromolecules in the regulation of eukaryotic cells. PMID- 19557511 TI - Polar lipid remodeling and increased sulfatide expression are associated with the glioma therapeutic candidates, wild type p53 elevation and the topoisomerase-1 inhibitor, irinotecan. AB - We report changes in gene and polar lipid expression induced by adenovirus delivered wild-type (wt) p53 gene and chemotherapy of U87 MG glioblastoma cells, a treatment known to trigger apoptosis and cell cycle arrest. Sulfatides (sulfonated glycolipids) were most highly modulated by wild-type p53 treatment; however, no changes were observed in expression levels of mRNA for genes involved in sulfatide metabolism, indicating post-transcriptional control of sulfatide synthesis. Modulation of the aglycones of GD1 and GM1b was observed in wild-type p53-treated cells. The treatment also leads to an increase in phospholipids such as phosphatidyl inositols, phosphatidyl serines, phosphatidyl glycerols, and phosphatidyl ethanolamines, especially hydroxylated phospholipids. These dramatic changes in the composition of cellular glycolipids in response to p53 gene expression and cytotoxic chemotherapy treatment indicate the large role that they play in cell signaling. The use of the human glioma cell line U87 appears to be an excellent model system both in tissue culture and in intracranial murine xenograft models to further characterize the role of sulfatides in modulating glioma responsivity to therapeutic agents. PMID- 19557513 TI - Neurite outgrowth effect of 4-O-methylhonokiol by induction of neurotrophic factors through ERK activation. AB - Compounds isolated from Magnolia officinalis such as magnolol, honokiol and obovatol exhibit several pharmacological effects on CNS including depressant, anxiolytic and anticonvulsant effects, as well as neuroprotective effects against chemical and heat damages. Recently, honokiol was found to have a neurotrophic effect in fetal rat cortical neurons. In the present study, we show that 4-O methylhonokiol, a novel compound from Magnolia officinalis, promotes neurite outgrowth in a concentration- dependent manner in rat embryonic neuronal cells. In parallel with the neurite outgrowth activity, the expression of neurite outgrowth marker proteins is also increased by treatment with 4-O-methylhonokiol. We also found that 4-O-methylhonokiol promotes the release of NGF and BDNF into cell culture medium. In addition, lower concentration of 4-O-methylhonokiol (1 and 2 lM) further enhanced neurite outgrowth and expression of neurite outgrowth marker proteins in the presence of NGF (50 ng/ml) or BDNF (10 ng/ml). Subsequently, we found that 4-O-methylhonokiol activates ERK in a concentration- dependent manner. However, the neurite outgrowth activity and the NGF and BDNF release induced by 4-O-methylhonokiol are suppressed by an ERK-specific inhibitor. These results suggest that 4-O-methylhonokiol has the ability to induce neurite outgrowth via the increase of neurotrophic factor levels through ERK activation. PMID- 19557514 TI - Oligodendrocytes are a novel source of amyloid peptide generation. AB - Alzheimer's disease is characterised by regional neuronal degeneration, synaptic loss, and the progressive deposition of the 4 kDa beta-amyloid peptide (Abeta) in senile plaques and accumulation of tau protein as neurofibrillary tangles. Abeta derives from the larger precursor molecule, amyloid precursor protein (APP) by proteolytic processing via beta- and gamma-secretases. While APP expression is well documented in neurons and astrocytes, the case for oligodendrocytes is less clear. The latter cell type is reported to express different isoforms of APP, and we have confirmed this observation by immunocytochemistry in cultures of differentiated rat cortical oligodendrocytes. Moreover, by means of a sensitive electrochemiluminescent immunoassay employing Abeta C-terminal specific antibodies, mature oligodendrocytes are shown to secrete the 40 and 42 amino acid Abeta species (Abeta40 and Abeta42). Secretion of Abeta peptides was reduced by incubating oligodendrocytes with alpha- and beta-secretase inhibitors, or a gamma secretase inhibitor. Disturbances of APP processing and/ or synthesis in oligodendrocytes may account for some myelin disorders observed in Alzheimer's disease and other senile dementias. PMID- 19557515 TI - Sodium metabisulfite modulation of potassium channels in pain-sensing dorsal root ganglion neurons. AB - The effects of sodium metabisulfite (SMB), a general food preservative, on potassium currents in rat dorsal root ganglion (DRG) neurons were investigated using the whole-cell patch-clamp technique. SMB increased the amplitudes of both transient outward potassium currents and delayed rectifier potassium current in concentration- and voltage-dependent manner. The transient outward potassium currents (TOCs) include a fast inactivating (A-current or IA) current and a slow inactivating (D-current or ID) current. SMB majorly increased IA, and ID was little affected. SMB did not affect the activation process of transient outward currents (TOCs), but the inactivation curve of TOCs was shifted to more positive potentials. The inactivation time constants of TOCs were also increased by SMB. For delayed rectifier potassium current (IK), SMB shifted the activation curve to hyperpolarizing direction. SMB differently affected TOCs and IK, its effects major on A-type K+ channels, which play a role in adjusting pain sensitivity in response to peripheral redox conditions. SMB did not increase TOCs and IK when adding DTT in pipette solution. These results suggested that SMB might oxidize potassium channels, which relate to adjusting pain sensitivity in pain-sensing DRG neurons. PMID- 19557516 TI - Frequency and risk factors for Barrett's esophagus in Taiwanese patients: a prospective study in a tertiary referral center. AB - BACKGROUND: There is a paucity of epidemiologic data concerning Barrett's esophagus (BE) in Taiwan. AIM: This study aimed to investigate the frequency of and risk factors for BE in self-referred Taiwanese patients undergoing diagnostic endoscopy. METHODS: A total of 736 consecutive patients undergoing upper endoscopy for a variety of gastro-intestinal symptoms from February to October 2007 were evaluated. A standard questionnaire was used to record the clinical characteristics and patient symptoms. Gastro-esophageal reflux disease (GERD) was diagnosed using the Montreal definition, while the Los Angeles Classification and Prague Circumferential and Maximal Criteria were used to assess erosive esophagitis and BE, respectively. Four-quadrant biopsies were taken from endoscopically suspected esophageal metaplastic mucosa every 2 cm for histologic evaluation. Eight variables were tested using a logistic regression model to identify risk factors for BE in GERD patients. RESULTS: GERD was diagnosed in 344 patients, with typical esophageal symptoms noted in 255, reflux chest pain syndrome in 107, and extra-esophageal syndrome in 51, while 27 were asymptomatic. The mean age of the GERD patients was 49.8 years and 55.5% of them were male. Thirty-six percent (123 of 344) demonstrated erosive esophagitis and 95% were classified as having Los Angeles grade A or B disease. BE was diagnosed in 13 patients (3.8% of GERD patients), three of whom had dysplastic mucosa. In the final analysis model, hiatal hernia (odds ratio [OR] = 4.7, 95% confidence interval [CI] = 1.3-17.7, P = 0.02) and GERD duration >5 years (OR = 4.2, 95% CI = 1.2-4.8, P = 0.03) were independent risk factors for the development of BE. CONCLUSION: There is a 3.8% frequency of BE in Taiwanese GERD patients. Hiatal hernia and prolonged GERD duration are significant risk factors. PMID- 19557517 TI - Rabeprazole 20 mg for erosive esophagitis-associated symptoms in a large, community-based study: additional results. AB - PURPOSE: To provide additional efficacy data in patients treated with rabeprazole through week 4, and to validate sustained relief of gastroesophageal reflux disease symptoms through week 8 as well as to further analyze rabeprazole safety in patients with wide-ranging demographic and clinical characteristics. RESULTS: Patients in this study (N = 2,449) demonstrated significant overall improvement versus baseline (P < 0.001). Substantial symptom relief was seen throughout 8 weeks of treatment. By week 4, complete relief of daytime and nighttime heartburn, belching, regurgitation, and dysphagia was observed in 87.5, 90.7, 50.7, 77.6, and 75.1% of patients, respectively. Improvements were seen in rabeprazole-treated patients (<65 or >or=65 years) with a range of baseline symptom severities and across different racial groups. Rabeprazole was well tolerated. CONCLUSIONS: In patients with endoscopy-confirmed erosive esophagitis treated with once-daily rabeprazole 20 mg, prompt and continuing improvements were seen in daytime and nighttime heartburn, belching, regurgitation, and dysphagia. PMID- 19557518 TI - Secular changes in mortality disparities in New York City: a reexamination. AB - Previously published analyses showed that inequalities in mortality rates between residents of poor and wealthy neighborhoods in New York City (NYC) narrowed between 1990 and 2000, but these trends may have been influenced by population in migration and gentrification. The NYC public housing population has been less subject to these population shifts than those in other NYC neighborhoods. We compared changes in mortality rates (MRs) from 1989-1991 to 1999-2001 among residents of NYC census blocks consisting entirely of public housing residences with residents of nonpublic housing low-income and higher-income blocks. Public housing and nonpublic housing low-income blocks were those in census block groups with > or =50% of residents living at <1.5 times the federal poverty level (FPL); nonpublic housing higher-income blocks were those in census block groups with <50% of residents living at <1.5 times the FPL. Information on deaths was obtained from NYC's vital registry, and US Census data were used for denominators. Age-standardized all-cause MRs in public housing, low-income, and higher-income residents decreased between the decades by 16%, 28%, and 22%, respectively. While mortality rate ratios between low-income and higher-income residents narrowed by 8%, the relative disparity between public housing and low income residents widened by 21%. Diseases amenable to prevention including malignancies, diabetes, and chronic lung disease contributed to the increased overall mortality disparity between public housing and lower-income residents. These findings temper previous findings that inequalities in the health of poor and wealthier NYC neighborhood residents have narrowed. NYC public housing residents should be a high-priority population for efforts to reduce health disparities. PMID- 19557519 TI - The impact of disease progression on perceived health status and quality of life of long-term cancer survivors. AB - INTRODUCTION: The number of cancer survivors experiencing disease progression (DP) is increasing with the number of cancer survivors. However, little is known whether DP affects health-related quality of life (HRQL) of long-term cancer survivors. We aimed therefore to compare the health status (HS) and HRQL of DP and disease-free (DF) survivors up to 15 years after initial diagnosis. METHODS: 232 cancer survivors with DP identified through the Eindhoven Cancer Registry were matched with 232 DF survivors of similar demographic and clinical characteristics. Patients completed generic HS (SF-36) and cancer-specific HRQL (QOL-CS) questionnaires 5-15 years after diagnosis. RESULTS: Compared with DF survivors, DP survivors exhibited significantly lower scores on all SF-36 and QOL CS (except spiritual well-being) dimensions. DF survivors had better scores than the normative population on all SF-36 dimensions. Among survivors with DP, those with short survival (<5 years) had significantly poorer HS scores on all dimensions except bodily pain compared with the normative population. Comparatively, the long survival (>or=5 years) DP group had better HRQL than the short DP group but poorer HRQL than the normative population. In multivariate analyses, DP and DF survival time were independently associated with aspects of HS and HRQL in cancer survivors. DISCUSSIONS/CONCLUSIONS: DP cancer survivors have poorer long-term HS and HRQL compared with DF survivors. However, there is suggestion that HS and HRQL does improve over time following DP. IMPLICATION FOR CANCER SURVIVORS: Although DP survivors report poorer long-term HRQL compared with DF cancer survivors, results suggest that time can attenuate the distress of DP on HRQL. Psycho-educational programs could help to increase patients' sense of empowerment and personal control should DP occur. PMID- 19557520 TI - Numerical design of RN(n)(nu) symmetry-based RF pulse schemes for recoupling and decoupling of nuclear spin interactions at high MAS frequencies. AB - An approach for the efficient implementation of RN(n)(nu) symmetry-based pulse schemes that are often employed for recoupling and decoupling of nuclear spin interactions in biological solid state NMR investigations is demonstrated at high magic-angle spinning frequencies. RF pulse sequences belonging to the RN(n)(nu) symmetry involve the repeated application of the pulse sandwich {R(phi)R(-phi)}, corresponding to a propagator U(RF) = exp(-i4phiI(z)), where phi = pinu/N and R is typically a pulse that rotates the nuclear spins through 180 degrees about the x-axis. In this study, broadband, phase-modulated 180 degrees pulses of constant amplitude were employed as the initial 'R' element and the phase-modulation profile of this 'R' element was numerically optimised for generating RN(n)(nu) symmetry-based pulse schemes with satisfactory magnetisation transfer characteristics. At representative MAS frequencies, RF pulse sequences were implemented for achieving 13C-13C double-quantum dipolar recoupling and through bond scalar coupling mediated chemical shift correlation and evaluated via numerical simulations and experimental measurements. The results from these investigations are presented here. PMID- 19557522 TI - Influence of sulfate reduction on the microbial dechlorination of pentachloroaniline in a mixed anaerobic culture. AB - The reductive dechlorination of pentachloroaniline (PCA) was investigated in the absence and presence of sulfate in batch assays using a PCA-dechlorinating mixed anaerobic culture with methanol as the external electron donor at neutral pH and 22 degrees C. PCA at an initial concentration of 7.8 microM was sequentially dechlorinated to dichlorinated anilines in the sulfate-free culture and the culture amended with 300 mg sulfate-S/L. At an initial concentration of 890 mg sulfate-S/L, a higher sulfate reduction rate was achieved, but PCA dechlorination was not observed until the sulfate concentration dropped to about 100 mg S/L. The transient inhibition of PCA is attributed to competition between sulfate reducing and dechlorinating species for electron donor, more likely for H(2) resulting from methanol fermentation. A long-term (118 days) PCA dechlorination assay with the sulfate-amended culture, which included five feeding cycles, resulted in accumulation of both sulfide (886 mg S/L) and acetate (1,900 mg COD/L). Under these conditions, the sulfate reducers were inhibited, while the rate and pathway of PCA dechlorination were not affected. The results of this study show that the rate of sulfate reduction rather than the sulfate concentration alone dictates the outcome of the competition between sulfate reducers and either dechlorinators or methanogens. The findings of the present study have significant implications relative to the fate and transport of PCA and its dechlorination products in sulfate-laden subsurface systems. PMID- 19557523 TI - Biodegradation kinetics of methyl iso-butyl ketone by acclimated mixed culture. AB - Methyl iso-butyl ketone (MIBK) is a widely used volatile organic compound (VOC) which is highly toxic in nature and has significant adverse effects on human beings. The present study deals with the removal of MIBK using biodegradation by an acclimated mixed culture developed from activated sludge. The biodegradation of MIBK is studied for an initial MIBK concentration ranging from 200-700 mg l( 1) in a batch mode of operation. The maximum specific growth rate achieved is 0.128 h(-1) at 600 mg l(-1)of initial MIBK concentration. The kinetic parameters are estimated using five growth kinetic models for biodegradation of organic compounds available in the literature. The experimental data found to fit well with the Luong model (R(2) = 0.904) as compared to Haldane model (R(2) = 0.702) and Edward model (R(2) = 0.786). The coefficient of determination (R(2)) obtained for the other two models, Monod and Powell models are 0.497 and 0.533, respectively. The biodegradation rate found to follow the three-half-order kinetics and the resulting kinetic parameters are reported. PMID- 19557524 TI - A new concept for reduction of diffuse contamination by simultaneous application of pesticide and pesticide-degrading microorganisms. AB - Pesticide residues and their transformation products are frequently found in groundwater and surface waters. This study examined whether adding pesticide degrading microorganisms simultaneously with the pesticide at application could significantly reduce diffuse contamination from pesticide use. Degradation of the phenoxyacetic acid herbicides MCPA (4-chloro-2-methylphenoxyacetic acid) and 2,4 D (2,4-dichlorophenoxyacetic acid) was studied in soil microcosm experiments after simultaneous spraying of herbicide and herbicide-degrading bacteria on an agricultural soil and on a sand with low degradation potential. The latter represented pesticide use on non-agricultural soils poor in microbial activity. Degradation and possible loss of herbicidal effect were also tested in a system with plants and the amounts of bacteria needed to give satisfactory MCPA degradation rate and the survival of degrading bacteria in formulated MCPA were determined. The results showed >80-99% degradation of 2,4-D and MCPA in soil within 1 day and >99% within 3 days after inoculation with 10(5)-10(7) herbicide degrading bacteria g(-1) dry weight of soil. Enhanced degradation of MCPA was also obtained in the presence of winter wheat and white mustard without loss of the intended herbicidal effect on white mustard. The survival of an isolated MCPA degrading Sphingomonas sp. in three realistic concentrations of formulated MCPA was very poor, showing that in practical applications direct contact between the microorganisms and the pesticide formulation must be precluded. The applicability and economic feasibility of the method and the information needed to obtain a useable product for field use are discussed. PMID- 19557521 TI - Parallel pigment and transcriptomic analysis of four barley albina and xantha mutants reveals the complex network of the chloroplast-dependent metabolism. AB - We investigated the pigment composition and the transcriptome of albina (alb-e ( 16 ) and alb-f ( 17 )) and xantha (xan-s ( 46 ) and xan-b ( 12 )) barley mutants to provide an overall transcriptional picture of genes whose expression is interconnected with chloroplast activities and to search for candidate genes associated with the mutations. Beside those encoding plastid-localized proteins, more than 3,000 genes involved in non-chloroplast localized metabolism were up /down-regulated in the mutants revealing the network of chloroplast-dependent metabolic pathways. The alb-e ( 16 ) mutant was characterized by overaccumulation of protoporphyrin IX upon ALA (5-amino levulinic acid) feeding and down regulation of the gene encoding one subunit of Mg-chelatase, suggesting a block of the chlorophyll biosynthetic pathway before Mg-protoporphyrin IX biosynthesis, while alb-f ( 17 ) overaccumulated Mg-protoporphyrin IX and repressed PorA expression, without alterations in Mg-chelatase mRNA level. The alb-f ( 17 )mutant also showed overexpression of several genes involved in phytochrome and in phytochrome-dependent pathways. The results indicate that the down-regulation of Lhcb genes in alb-e ( 16 ) cannot be mediated by the accumulation of Mg protoporphyrin IX. After ALA treatment, xan-s ( 46 ) showed overaccumulation of Mg-protoporphyrin IX, while the relative porphyrin composition of xan-b ( 12 ) was similar to wild type. The transcripts encoding the components of several mitochondrial metabolic pathways were up-regulated in albina/xantha leaves to compensate for the absence of active chloroplasts. The mRNAs encoding gun3, gun4, and gun5 barley homologous genes showed significant expression variations and were used to search for co-expressed genes across all samples. These analyses provide additional evidences on a chloroplast-dependent covariation of large sets of nuclear genes. PMID- 19557525 TI - Incidence and prognosis of cholangiocarcinoma in Danish patients with and without inflammatory bowel disease: a national cohort study, 1978-2003. AB - Patients with inflammatory bowel disease (IBD) are at increased risk of cholangiocarcinoma (CC), but quantitative data are scant. Furthermore, little is known about the impact of IBD on CC occurrence and prognosis. Based on nationwide population-based registries we compared the incidence and survival of CC patients with and without IBD from 1978 to 2003. We used the National Registry of Patients and the Danish Cancer Registry to identify patients with IBD and CC. From the Civil Registration System we identified population controls. We calculated incidence rates, incidence rate ratios (compared with population controls), and absolute cumulative risks. We also computed median survival in CC patients with and without IBD. 2,725 CC patients were identified. The incidence of CC among the 41,280 IBD patients was 7.6 per 100,000 person years compared with 1.9 per 100,000 among the 412,796 population controls (four-fold increased risk). The 10 year cumulative risk of CC in IBD patients was 0.07%. Sub analyses showed that the increased risk of CC was more pronounced in male IBD patients and in patients with ulcerative colitis. We found a decreasing CC incidence in IBD patients over calendar time. CC patients with IBD were, on average, 15 years younger at cancer diagnosis than IBD-free CC patients, and median survival was 1 month in both groups. In conclusion, the absolute risk of CC in IBD patients was low and the CC incidence decreased over calendar time. The prognosis was equally grave, regardless of the presence of IBD. PMID- 19557526 TI - The association between birthplace in different regions of the world and cardiovascular mortality among residents of Spain. AB - The objective of this study is to estimate the association of birthplace with mortality from cardiovascular diseases in residents of Spain by analysing immigrant populations that are unlikely to have adopted health-related attitudes and behaviours of the host country. Data from the population register and cause of death register were used for the period 2001-2005. The study included people aged 20-64 years. Age-adjusted mortality from cardiovascular diseases--and from ischaemic heart disease, cerebrovascular disease and hypertension-related disease -according to birthplace were estimated and compared with those for the native Spanish population by mortality rate ratios. Compared with the native Spanish population, residents who came from Eastern Europe, Sub-Saharan Africa and South Asia had a higher risk of mortality from most of the cardiovascular diseases analysed. Women from North Africa and the Caribbean also had a higher risk of cardiovascular mortality. A higher risk of mortality from ischemic heart disease was observed in persons from the Middle East, and from cerebrovascular disease in those from Eastern Asia. Compared with the native Spanish population, residents from South America and Eastern Asia had a lower risk of mortality from ischemic heart disease. This pattern of mortality from cardiovascular diseases in residents of Spain who have come from different regions of the world is very similar to the findings of studies in other countries, and probably reflects the burden of disease in the countries of origin. PMID- 19557528 TI - Megnin re-analysed: the case of the newborn baby girl, Paris, 1878. AB - This paper analyses acarological evidence from a 130-year-old forensic investigation. It was the first case in forensic acarology, i.e., the first case where mites provided substantial information to estimate the post-mortem interval (PMI). In 1878, the mites found in the mummified body of a newborn baby girl in Paris, France, were studied by acarologist and forensic entomologist Jean Pierre Megnin. Megnin estimated around 2.4 million mites in the skull and identified them as Tyroglyphus longior (Gervais), a junior synonym of Tyrophagus longior. He suggested that the arrival of these mites at the corpse would have occurred by phoresy on carrier insects, roughly 5 months before the autopsy. There is no doubt about the identification of the mites, Megnin was a highly respected acarologist. However, two main factors affecting the biology of Tyrophagus mites were not included in the original analysis. First, Megnin stated that the mites were phoretic. However, he probably did not have access to information about the natural history of the species, because as a rule Tyrophagus mites are non phoretic. Considering the omnipresence of Tyrophagus mites in soil, most likely the mites will have arrived almost immediately after death. Second, temperature was not taken into account during the estimations of the mite population growth rate. The new analysis is based on current knowledge of Tyrophagus biology and includes temperature, estimated following a handful of weather reports of the years 1877 and 1878. The new projections indicate that non-phoretic mites may have colonised the body just after death and the colony would have built up over 8 months, contrary to the 5 months proposed by Megnin. This new lapse of time agrees with the PMI proposed by Brouardel: on 15 January 1878 he postulated the death of the newborn to have occurred some 8 months before the autopsy. PMID- 19557527 TI - Phoretic mites associated with animal and human decomposition. AB - Phoretic mites are likely the most abundant arthropods found on carcases and corpses. They outnumber their scavenger carriers in both number and diversity. Many phoretic mites travel on scavenger insects and are highly specific; they will arrive on a particular species of host and no other. Because of this, they may be useful as trace indicators of their carriers even when their carriers are absent. Phoretic mites can be valuable markers of time. They are usually found in a specialised transitional transport or dispersal stage, often moulting and transforming to adults shortly after arrival on a carcase or corpse. Many are characterised by faster development and generation cycles than their carriers. Humans are normally unaware, but we too carry mites; they are skin mites that are present in our clothes. More than 212 phoretic mite species associated with carcases have been reported in the literature. Among these, mites belonging to the Mesostigmata form the dominant group, represented by 127 species with 25 phoretic mite species belonging to the family Parasitidae and 48 to the Macrochelidae. Most of these mesostigmatids are associated with particular species of flies or carrion beetles, though some are associated with small mammals arriving during the early stages of decomposition. During dry decay, members of the Astigmata are more frequently found; 52 species are phoretic on scavengers, and the majority of these travel on late-arriving scavengers such as hide beetles, skin beetles and moths. Several species of carrion beetles can visit a corpse simultaneously, and each may carry 1-10 species of phoretic mites. An informative diversity of phoretic mites may be found on a decaying carcass at any given time. The composition of the phoretic mite assemblage on a carcass might provide valuable information about the conditions of and time elapsed since death. PMID- 19557529 TI - Human hair follicle mites and forensic acarology. AB - The hair follicle mites of the genus Demodex (Demodecidae) were first discovered in humans in 1841. Since then, members of this host-specific genus have been found in 11 of the 18 orders of eutherian mammals with most host species harboring two or more species of Demodex. Humans are host to D. folliculorum and D. brevis. The biology, natural history, and anatomy of these mites as related to their life in the human pilosebaceous complex is reviewed. This information may provide insight into the application of Demodex as a tool for the forensic acarologist/entomologist. PMID- 19557530 TI - Functional blocking of specific integrins inhibit colonic cancer migration. AB - For more effective oncological management of disseminated colorectal cancer, therapies must be devised that target the different individual stages of metastasis development. Recent work showed that integrin subunits alpha2, alpha6 and beta4 are involved in the colorectal cancer cell extravasation process. By means of Immunocytochemistry and Western blotting, it was shown that all three integrins are expressed not only in human colorectal cancer cells (HT29) but also in rat colonic cancer cells (DHDK12). Using in vivo models and intravital video microscopy techniques, it was shown that functional blocking of these integrin subunits by specific antibodies produced a significant reduction in cancer cell extravasation and migration. In conclusion, integrin subunits alpha2, alpha6 and beta4 are expressed in unrelated colorectal cancer cell strains and appear to play a key role in cancer cell migration. PMID- 19557532 TI - Water quality status of dugouts from five districts in Northern Ghana: implications for sustainable water resources management in a water stressed tropical savannah environment. AB - This study was primarily aimed at investigating the physicochemical and microbial quality of water in 14 such dugouts from five districts in the northern region of Ghana. Results obtained suggest that except for colour, turbidity, total iron and manganese, many physicochemical parameters were either within or close to the World Health Organisation's acceptable limits for drinking water. Generally, colour ranged from 5 to 750 Hz (mean 175 Hz), turbidity from 0.65 to 568 nephelometric turbidity units (NTU; mean 87.9 NTU), total iron from 0.07 to 7.85 mg/L (mean 1.0 mg/L) and manganese from 0.03 to 1.59 mg/L (mean 0.50 mg/L). Coliform counts in water from all the dugouts in both wet and dry seasons were, however, above the recommended limits for drinking water. Total and faecal coliforms ranged from 125 to 68,000 colony forming units (cfu)/100 mL (mean 10,623 cfu/100 mL) and <1 to 19,000 cfu/100 mL (mean 1,310 cfu /100 mL), respectively. The poor microbial quality, as indicated by the analytically significant presence of coliform bacteria in all samples of dugout water, strongly suggests susceptibility and exposure to waterborne diseases of, and consequent health implications on, the many people who continuously patronise these vital water resources throughout the year. In particular, more proactive sustainable water management options, such as introduction to communities of simple but cost-effective purification techniques for water drawn from dugouts for drinking purposes, education and information dissemination to the water users to ensure environmentally hygienic practices around dugouts, may be needed. PMID- 19557531 TI - Disparities in survival after Hodgkin lymphoma: a population-based study. AB - Survival after Hodgkin lymphoma (HL) is generally favorable, but may vary by patient demographic characteristics. The authors examined HL survival according to race/ethnicity and neighborhood socioeconomic status (SES), determined from residential census-block group at diagnosis. For 12,492 classical HL patients >= 15 years diagnosed in California during 1988-2006 and followed through 2007, we determined risk of overall and HL-specific death using Cox proportional hazards regression; analyses were stratified by age and Ann Arbor stage. Irrespective of disease stage, patients with lower neighborhood SES had worse overall and HL specific survival than patients with higher SES. Patients with the lowest quintile of neighborhood SES had a 64% (patients aged 15-44 years) and 36% (>= 45 years) increased risk of HL-death compared to patients with the highest quintile of SES; SES results were similar for overall survival. Even after adjustment for neighborhood SES, blacks and Hispanics had increased risks of HL-death 74% and 43% (15-44 years) and 40% and 17% (>= 45 years), respectively, higher than white patients. The racial/ethnic differences in survival were evident for all stages of disease. These data provide evidence for substantial, and probably remediable, racial/ethnic and neighborhood SES disparities in HL outcomes. PMID- 19557534 TI - A comparison of storm-based and annual-based indices of hydrologic variability: a case study in Fort Benning, Georgia. AB - The magnitude, frequency, duration, timing, and rate of change of hydrologic conditions regulate ecological processes in aquatic ecosystems. Conditions are typically characterized using annual-based hydrologic indices derived from daily and/or monthly stream flow data. In this study, we present an alternative approach to identify hydrologic indices based on storm hydrographs. Hydrologic indices derived from long-term daily flow data were compared to those from storm events for two headwater watersheds in Fort Benning, Georgia. Five hydrologic indices derived from daily flow data and storm events shared common features. Storm-based magnitude of mean peak discharge and mean response factor, frequency of bankfull discharge, rate of change in mean slopes of rising, and falling limb of the hydrograph were consistent with the results from long-term daily flow data. The annual flow increases and decreases were well matched by stormflow rising and falling. Both indicators showed one watershed having three times the response rates as compared to the other. Results suggested that select storm based indices may be used as surrogates to the indices derived from long-term data. PMID- 19557533 TI - Cadmium partition in river sediments from an area affected by mining activities. AB - In this paper, the cadmium distribution in Certej River sediments in an area seriously affected by intense mining activities has been studied. The main objective of this study was the evaluation of partition of this metal into different operational defined fractions by sequential extractions. Community Bureau of Reference (BCR) sequential extraction was used to isolate different fractions. The sediment quality was assessed both upstream and downstream the pollution input points, along the Certej River, in order to reveal a possible accumulation of cadmium in sediments and the seasonal changes in cadmium concentrations in BCR sediment phases. Our results reveal that most of the cadmium content is divided between both the soluble and iron and manganese hydrated oxide fractions. Based on total cadmium concentrations in sediments, the enrichment factors were estimated using aluminum as normalizing element and the regression curve Cd/Al corresponding to the geochemical background of the studied area. PMID- 19557535 TI - Dry deposition velocity of atmospheric nitrogen in a typical red soil agro ecosystem in Southeastern China. AB - Atmospheric dry deposition is an important nitrogen (N) input to farmland ecosystems. The main nitrogen compounds in the atmosphere include gaseous N (NH3, NO2, HNO3) and aerosol N (NH4+/NO3-). With the knowledge of increasing agricultural effects by dry deposition of nitrogen, researchers have paid great attention to this topic. Based on the big-leaf resistance dry deposition model, dry N deposition velocities (Vd) in a typical red soil agro-ecosystem, Yingtan, Jiangxi, Southeastern China, were estimated with the data from an Auto Meteorological Experiment Station during 2004-2007. The results show that hourly deposition velocities (Vdh) were in the range of 0.17-0.34, 0.05-0.24, 0.57-1.27, and 0.05-0.41 cm/s for NH3, NO2, HNO3, and aerosol N, respectively, and the Vdh were much higher in daytime than in nighttime and had a peak value around noon. Monthly dry deposition velocities (Vdm) were in the range of 0.14-0.36, 0.06 0.18, and 0.07-0.25 cm/s for NH3, NO2, and aerosol N, respectively. Their minimum values appeared from June to August, while their maximum values occurred from February to March each year. The maximum value for HNO3 deposition velocities appeared in July each year, and Vdm(HNO3) ranged from 0.58 to 1.31 cm/s during the 4 years. As for seasonal deposition velocities (Vds), Vds(NH3), Vds(NO2), and Vds(aerosol N) in winter or spring were significantly higher than those in summer or autumn, while Vds(HNO3) in summer were higher than that in winter. In addition, there is no significant difference among all the annual means for deposition velocities (Vda). The average values for NH3, NO2, HNO3, and aerosol N deposition velocities in the 4 years were 0.26, 0.12, 0.81, and 0.16 cm/s, respectively. The model is convenient and feasible to estimate dry deposition velocity of atmospheric nitrogen in the typical red soil agro-ecosystem. PMID- 19557536 TI - Inhibitors of adenosine consuming parasites through polymer-assisted N-acylation of N6-substituted 5'-amino-5'-deoxyadenosines. AB - A series of 30 adenosine derivatives with three different substituents at the N(6)-position were prepared in order to evaluate their potential to inhibit the pathogenic protozoa Plasmodium falciparum and Trypanosoma brucei in vitro. The rationale for synthesis of these structures was the high probability of interactions with multiple adenosine associated targets and the assumption that N(6)-substitutents should increase stability against adenosine deaminases and allow the molecules to diffuse across parasite membranes. Starting from inosine, the new compounds were prepared as single isomers using a polymer-assisted acylation protocol enabling the straightforward isolation of the target compounds in pure form. Three of the compounds displayed anti-plasmodial and one anti trypanosomal activity in the single digit micromolar concentration range. PMID- 19557537 TI - Involvement of the cytoplasmic juxtamembrane region of matriptase in its exclusive localization to the basolateral membrane domain of Madin-Darby canine kidney epithelial cells. AB - Matriptase is a type II transmembrane serine protease. This protease is strongly expressed in simple epithelial cells such as enterocytes and kidney tubular cells in which the plasma membranes are separated into apical and basolateral domains. Although matriptase was found previously to occur exclusively on the basolateral membrane of enterocytes, the underlying mechanism of localization is unclear. In the present study, a full-length rat matriptase and a chimera consisting of the cytoplasmic and transmembrane regions of the protease and green fluorescent protein (designated as 1-86GFP) were found to localize exclusively to the basolateral membrane domain when expressed in Madin-Darby canine kidney epithelial cells. Mutagenesis analysis of 1-86GFP revealed that the matriptase cytoplasmic juxtamembrane amino acid residues (Lys45, Val47, and Arg50) play a role in mediating the localization in the cells. This study provides the first evidence that matriptase carries information for its localization in simple epithelia. PMID- 19557539 TI - Iron (III) citrate inhibits polyethylenimine-mediated transient transfection of Chinese hamster ovary cells in serum-free medium. AB - Recent advances in transient transfection protocols using polyethylenimine (PEI) as a transfection reagent have led to the development of economical methods that provide yields sufficient for industrial production of proteins for many preclinical needs. There are many variables that can be optimized to improve protein expression in transient transfection, and one of the most critical is the medium in which the cells are grown. While transfection with PEI works well in media containing serum, the biopharmaceutical industry is moving away from animal derived components in media. A number of serum-free media have been found to allow transient transfection, but many others do not for reasons that are not clear. Thus, knowledge of the components of serum-free media that can cause inhibition of PEI-mediated transient transfection would be useful for media development. In this study, an analysis was performed of various components of a serum-free medium used for Chinese hamster ovary cells in which PEI-mediated transient transfection was inhibited. We found that an iron supplement added to the medium was responsible for the inhibition. Further investigation showed that iron (III) citrate, a common iron chelator found in serum-free medium, was the specific component that caused the effect. Further, we showed that inhibition of transient transfection was caused by iron (III) citrate specifically, rather than citrate or iron alone. Finally, we showed that various iron chelators in serum free media other than iron (III) citrate do not inhibit antibody expression. PMID- 19557538 TI - 5-Azacytidine facilitates osteogenic gene expression and differentiation of mesenchymal stem cells by alteration in DNA methylation. AB - Mesenchymal stem cells (MSCs) are considered to be one of the most promising therapeutic cell sources as they encompass a plasticity of multiple cell lineages. The challenge in using these cells lies in developing well-defined protocols for directing cellular differentiation to generate a desired lineage. In this study, we investigated the effect of 5-azacytidine, a DNA demethylating agent, on osteogenic differentiation of MSCs. The cells were exposed to 5 azacytidine in culture medium for 24 h prior to osteogenic induction. Osteogenic differentiation was determined by several the appearance of a number of osteogenesis characteristics, including gene expression, ALP activity, and calcium mineralization. Pretreatment of MSCs with 5-azacytidine significantly facilitated osteogenic differentiation and was accompanied by hypomethylation of genomic DNA and increased osteogenic gene expression. Taking dlx5 as a representative, methylation alterations of the "CpG island shore" in the promoter caused by 5-azacytidine appeared to contribute to osteogenic differentiation. PMID- 19557540 TI - Detailed characterization of Mirafiori lettuce virus-resistant transgenic lettuce. AB - Lettuce big-vein disease is caused by Mirafiori lettuce virus (MiLV), which is vectored by the soil-borne fungus Olpidium brassicae. A MiLV-resistant transgenic lettuce line was developed through introducing inverted repeats of the MiLV coat protein (CP) gene. Here, a detailed characterization study of this lettuce line was conducted by comparing it with the parental, non-transformed 'Kaiser' cultivar. There were no significant differences between transgenic and non transgenic lettuce in terms of pollen fertility, pollen dispersal, seed production, seed dispersal, dormancy, germination, growth of seedlings under low or high temperature, chromatographic patterns of leaf extracts, or effects of lettuce on the growth of broccoli or soil microflora. A significant difference in pollen size was noted, but the difference was small. The length of the cotyledons of the transgenic lettuce was shorter than that of 'Kaiser,' but there were no differences in other morphological characteristics. Agrobacterium tumefaciens used for the production of transgenic lettuce was not detected in transgenic seeds. The transgenic T(3), T(4), and T(5) generations showed higher resistance to MiLV and big-vein symptoms expression than the resistant 'Pacific' cultivar, indicating that high resistance to lettuce big-vein disease is stably inherited. PCR analysis showed that segregation of the CP gene was nearly 3:1 in the T(1) and T(2) generations, and that the transgenic T(3) generation was homozygous for the CP gene. Segregation of the neomycin phosphotransferase II (npt II) gene was about 3:1 in the T(1) generation, but the full length npt II gene was not detected in the T(2) or T(3) generation. The segregation pattern of the CP and npt II genes in the T(1) generation showed the expected 9:3:3:1 ratio. These results suggest that the fragment including the CP gene and that including the npt II gene have been integrated into two unlinked loci, and that the T(1) plant selected in our study did not have the npt II gene. DNA sequences flanking T-DNA insertions in the T(2) generation were determined using inverse PCR, and showed that the right side of the T-DNA including the npt II gene had been truncated in the transgenic lettuce. PMID- 19557541 TI - Molecular phylogenetic studies on Theileria spp. isolates (China) based on small subunit ribosomal RNA gene sequences. AB - Six Theileria spp. from cattle, buffalo and black goat were identified in the Hubei province of China. In order to study the taxonomic status of these parasites, phylogenetic analysis of 18S rRNA genes were carried out. The 18S rRNA genes from each isolate were amplified by the polymerase chain reaction and the approximate 1.75 kb products were cloned and sequenced. Phylogenetic analysis of these gene sequences revealed that the five parasites from buffalo and cattle belonged to the Theileria sergenti/buffeli/orientalis group. The parasite from the Chinese goat (Macheng-Hubei, DQ286802) was closely related to Theileria luwenshuni isolated from sheep in the north of China. This represent the first report on the use of molecular phylogeny to classify Theileria spp. obtained in the Hubei province, showing that Theileria spp. from ruminants found in Hubei province belongs to the benign group of Theileria spp. PMID- 19557542 TI - Post-renal transplant surgical complications with newer immunosuppressive drugs: mycophenolate mofetil vs. m-TOR inhibitors. AB - PURPOSE: To compare the incidence of immediate surgical complications after renal transplantation between mycophenolate mofetil (MMF group)-based and mTOR inhibitors (mTOR group)-based immunosuppressive regimens. METHODS: The preoperative parameters in the recipients, rejection rates and surgical complications within 12 months in the recipients were analyzed in 80 patients who had live related renal transplantation. The immunosuppressive regimen was based on MMF (MMF, prednisolone, cyclosporine) in 40 patients and mTOR inhibitors (sirolimus/everolimus, prednisolone, cyclosporine) in 40 patients. RESULTS: The baseline characteristics were comparable between the two groups. Infective complications (urinary tract infections, pulmonary infections and superficial wound infection) occurred in 27.5% (11/40) and 12.5% (5/40) of patients from MMF and mTORI groups, respectively (P = 0.096). Patients in mTORI group had significantly more wound dehiscence (8/40 i.e., 20%) than in MMF group (1/40 i.e., 2.5%) (P = 0.014). There was no significant difference in the occurrence of clinically significant or symptomatic lymphoceles that needed intervention (3 vs. 2). The hospital stay was significantly prolonged in mTORI group mainly because of wound-related problems (35 vs. 24 days). CONCLUSION: In the post-renal transplant setting, use of mTORI results in significantly higher wound complications compared to that of MMF leading to prolonged hospital stay. There is no significant difference in infective complications or lymphocele incidence between these two immunosuppressive regimens. PMID- 19557544 TI - Minimal genomes, maximal productivity: comparative genomics of the photosystem and light-harvesting complexes in the marine cyanobacterium, Prochlorococcus. AB - Although Prochlorococcus isolates possess the smallest genomes of any extant photosynthetic organism, this genus numerically dominates vast regions of the world's subtropical and tropical open oceans and has evolved to become an important contributor to global biogeochemical cycles. The sequencing of 12 Prochlorococcus genomes provides a glimpse of the extensive genetic heterogeneity and, thus, physiological potential of the lineage. In this study, we present an up-to-date comparative analysis of major proteins of the photosynthetic apparatus in 12 Prochlorococcus genomes. Our analyses reveal a striking diversity within the Prochlorococcus lineage in the major protein complexes of the photosynthetic apparatus. The heterogeneity that has evolved in the photosynthetic apparatus suggests versatility in strategies for optimizing photosynthesis under conditions of environmental variability and stress. This diversity could be particularly important in ensuring the survival of a lineage whose individuals have evolved minimal genomes and, thus, relatively limited repertoires for responding to environmental challenges. PMID- 19557545 TI - Autopsy case of Gaucher disease type I in a patient on enzyme replacement therapy. Comments on the dynamics of persistent storage process. AB - We report a female patient with Gaucher disease (GD) type I on ERT (imiglucerase) for 5 years, which led to a significant general improvement. Aged 59 years she underwent an episode of altitude sickness followed by sepsis, disseminated intravascular coagulation, and multiorgan failure. She succumbed to a cerebral haemorrhage. Autopsy revealed liver cholestatic cirrhosis and multifocal liver carcinoma with immunophenotype compatible with cholangiocarcinoma. Analysis of the storage process revealed its absence or very low levels in the majority of liver and spleen macrophages. Gaucher cells (GCs) were seen only as occasional aggregates of various sizes in these organs. GCs were seen also in the leptomeninx of the cerebellum and as infrequent perivascular clusters in both the grey and white cerebral matters. Bone marrow was heavily infiltrated with GCs, especially in the adipocyte-rich part. GCs in this location displayed varied degrees of cytoplasmic vacuolation unrelated to the lysosomal compartment, caused by droplets of triglyceride, and interpreted as due to resorption of fragments of altered white adipocytes. All these observations point to the relative efficacy of ERT in covering the standard substrate load, which should not be exceeded as it would lead to the evolution of mature GCs. The results are discussed in relation to our recently published hypothesis on GD cell pathology. PMID- 19557546 TI - Inhibitory effects of Ephedra major Host on Aspergillus parasiticus growth and aflatoxin production. AB - This study was undertaken to evaluate the effect of Ephedra major Host, an important medicinal plant with various biological activities, on growth and aflatoxin (AF) production by Aspergillus parasiticus NRRL 2999. The fungus was cultured in yeast extract-sucrose (YES) broth, a conductive medium that supports AF production, in the presence of various concentrations of essential oil (EO), hexanic and methanolic extracts of plant aerial parts, fruits, and roots using microbioassay technique. After incubating for 96 h at 28 degrees C in static conditions, mycelial dry weight was determined as an index of fungal growth, and aflatoxin B(1) (AFB(1)) was measured using HPLC technique. Based on the obtained results, EO of plant aerial parts significantly inhibited fungal growth at the highest concentration of 1000 microg/ml without any obvious effect on AFB(1) production at all concentrations used. Among plant extracts tested, only methanolic extract of aerial parts and roots were found to inhibit fungal growth and AFB(1) production dose-dependently with an IC(50) value of 559.74 and 3.98 microg/ml for AFB(1), respectively. Based on the GC/MS data, the major components of E. major EO were bis (2-ethylhexyl) phthalate (42.48%), pentacosane (20.94%), docosane (14.64%), citronellol (5.15%), heptadecan (4.41%), cis-3-Hexen-1-ol benzoate (4.07%), and 7-Octen-2-ol (3.25%). With respect to the potent inhibition of fungal growth and AF production by E. major, this plant may be useful in protecting crops from both toxigenic fungal growth and AF contamination. PMID- 19557547 TI - Circadian clock resetting in the mouse changes with age. AB - The most widely recognised consequence of normal age-related changes in biological timing is the sleep disruption that appears in old age and diminishes the quality of life. These sleep disorders are part of the normal ageing process and consist primarily of increased amounts of wakefulness and reduced amounts of deep sleep. Changes in the amplitude and timing of the sleep-wake cycle appear to represent, at least in part, a loss of effective circadian regulation of sleep. Understanding alterations in the characteristics of stimuli that help to consolidate internal rhythms will lead to recommendations to improve synchronisation in old age. Converging evidence from both human and animal studies indicate that senescence is associated with alterations in the neural structure thought to be primarily responsible for the generation of the circadian oscillation, the suprachiasmatic nuclei (SCN). Work has shown that there are changes in the anatomy, physiology and ability of the clock to reset in response to stimuli with age. Therefore it is possible that at least some of the observed age-related changes in sleep and circadian timing could be mediated at the level of the SCN. The SCN contain a circadian clock whose activity can be recorded in vitro for several days. We have tested the response of the circadian clock to a number of neurochemicals that reset the clock in a manner similar to light, including glutamate, N-methyl-D-aspartate (NMDA), gastrin-releasing peptide (GRP) and histamine (HA). In addition, we have also tested agents which phase shift in a pattern similar to behavioural 'non-photic' signals, including neuropeptide Y (NPY), serotonin (5HT) and gamma-aminobutyric acid (GABA). These were tested on the circadian clock in young and older mice (approximately 4 and 15 months old). We found deficits in the response to specific neurochemicals but not to others in our older mice. These results indicate that some changes seen in the responsiveness of the circadian clock to light with age may be mediated at the level of the SCN. Further, the responsiveness of the circadian clock with age is attenuated to some, but not all stimuli. This suggests that not all clock stimuli lose their effectiveness with age, and that it may be possible to compensate for deficits in clock performance by enhancing the strength of those stimulus pathways which are intact. PMID- 19557548 TI - The mechanism whereby heat shock induces apoptosis depends on the innate sensitivity of cells to stress. AB - The cellular response to heat shock (HS) is a paradigm for many human diseases collectively known as "protein conformation diseases" in which the accumulation of misfolded proteins induces cell death. Here, we analyzed how cells having a different apoptotic threshold die subsequent to a treatment with HS. Cells with a low apoptotic threshold mainly induced apoptosis through activation of conventional stress kinase signaling pathways. By contrast, cells with a high apoptotic threshold also died by apoptosis but likely after the accumulation of heat-aggregated proteins as revealed by the formation of aggresomes in these cells, which were associated with the generation of atypical nuclear deformations. Inhibition of the proteasome or expression of an aggregation prone protein produced similar nuclear alterations. Furthermore, elevated levels of chaperones markedly suppressed both HS-induced nuclear deformations and apoptosis induced upon protein aggregation whereas they had little effect on stress kinase mediated apoptosis. We conclude that the relative contribution of stress signaling pathways and the accumulation of protein aggregates to cell death by apoptosis is related to the innate sensitivity of cells to deadly insults. PMID- 19557549 TI - The possible role of isolated lymphoid follicles in colonic mucosal repair. AB - The continuous reformation and rapid repair of the colonic mucosa is essential for avoiding the aggregation of pernicious mutations induced by bacterial, toxic, or mitogenic factors. Gut-associated lymphoid tissue is supposed to play a central role in the organization of the repair mechanisms. In inflammatory conditions, the number, the diameter and the density of isolated lymphoid follicles (ILFs) are increasing. They are involved not just in immune surveillance, but their presence is also indispensable in normal mucosal regeneration of the colon. The relation of ILFs to the components of mucosal renewal such as bone marrow derived stem cells, follicular dendritic cells, subepithelial myofibroblasts or crypt formation has not been directly studied, and data about their putative organizer role are scattered in scientific literature. Whether they act as a regenerative pool containing stem cells in case of mucosal damage, or they are responsible only for the optimal cytokine milieu for the differentiation of immigrating stem cells is a question under debate. Our aim is to review the relation of ILFs to the different elements of colonic mucosal repair. PMID- 19557551 TI - Research on human-animal entities: ethical and regulatory aspects in Europe. AB - To review the ethical and regulatory issues related to the research on human animal entities at various stages. Review of scientific publications, laws and ethical guidelines in this field up through September 2008. The article presents the overall picture of the research on human-animal entities in Europe, including the public opinion and the country-specific regulations and guidelines regarding such research, discusses the ethical issues, including both arguments opposing and favoring such research, as well as discusses and clarifies the terminology used. Creation of human-animal entities with the potential for what may be viewed as human faculties raises profound questions concerning the rights and responsibilities of human beings. There is great need for informed discussions and interchanges between the expert researchers, ethicists, policymakers, lawyers and the public in general to come to consensus regarding the issues discussed in this paper. Suggestions regarding these rights and responsibilities are overviewed. PMID- 19557550 TI - Epileptic seizures in AD patients. AB - Epileptic seizures have long been recognised as a complication of the clinical syndrome of Alzheimer's disease, particularly in advanced disease, but have hitherto been viewed essentially as epiphenomena of the neurodegenerative process. Progress with animal models of Alzheimer's disease has suggested that this view may be incorrect, and that seizures may be a reflection of pathophysiological processes similar to or overlapping with those responsible for cognitive decline. This overlap between neuropsychological and neurophysiological changes suggests that seizures in Alzheimer's disease may be a valid therapeutic target, over and above symptomatic treatment. This article reviews data on the prevalence of seizures in Alzheimer's disease, seizure types, pathophysiology and treatment. Seizure prevalence increases with disease duration, but early-onset disease is associated with a greater risk of seizures, in part related to the frequency of presenilin-1 gene mutations in early-onset disease. Seizures are mostly of partial origin, with both complex partial and secondary generalised seizures. Seizure pathophysiology may relate to increased amyloid beta-peptide production, structural alterations in neurones related to cytoskeletal dysfunction, cerebrovascular changes, neurotransmitter dysfunction or combinations thereof. Through modification of these pathophysiological pathways, there may be possible roles for anti-epileptic drugs such as sodium valproate and lacosamide in the treatment of Alzheimer's disease. In summary, epileptic seizures are part of the AD phenotype, and merit further investigation. PMID- 19557552 TI - Incidence of FLT3 and nucleophosmin gene mutations in childhood acute myeloid leukemia: Serbian experience and the review of the literature. AB - Mutations in the fms-like tyrosine kinase 3 (FLT3) gene (internal tandem duplication (ITD) and point mutation in the tyrosine kinase domain, FLT3/D835) as well as the nucleophosmin (NPM1) gene are the most common abnormalities in adult acute myeloid leukemia (AML). Their significance in pediatric AML is still unclear. In this study we evaluated the frequency of FLT3 and NPM1 mutations in childhood AML. We also examined clinical features and outcome of these patients. FLT3 and NPM1 mutations were analysed in 42 and 37 childhood AML patients, respectively, using polymerase chain reaction (PCR) and direct sequencing. FLT3 mutations were detected in 4/42 patients (9.5%). The frequencies of FLT3/ITD and FLT3/D835 were the same, 2/42 (4.7%). NMP1 mutations were found in 1/37 patients (2.7%). FLT3 gene mutations were correlated with induction failure. Here we report the results of the study of FLT3 and NPM1 gene mutations in childhood AML patients in Serbia. Low frequencies of these molecular markers point out that these abnormalities are rare in this cohort of patients. Comparative study of data on NPM1 mutations in childhood AML revealed that various NPM1 gene mutation types are associated with childhood AML. Our findings as well as previously reported data, contributes to a hypothesis of different biology and etiology of adult and childhood AML. More extensive studies of NPM1 and FLT3 mutations in childhood AML are needed to determine their biological and clinical importance. PMID- 19557553 TI - Use of criterion-based clinical audit to improve the quality of obstetric care: A systematic review. AB - Although there is evidence that audit and feedback can improve health outcomes, little is known about the effect of audit on the quality of care from client's perspective. The aim of the current review was to explore the use of criterion based audit to improve quality of obstetric care from both the midwives/doctors' and women/mothers' perspectives. Electronic searches of Cochrane Library, MEDLINE, and EMBASE were conducted. Randomized controlled trials (RCTs) and before-and-after studies that assessed the effect of criterion-based audit on health outcomes or women/mothers' perception of obstetric care were set as selection criteria. Data were extracted, analyzed using Revman 4.2 software, and results expressed as weighted mean differences for continuous data, and odds ratios for dichotomous data. Nineteen studies (one RCT and 18 before-and-after studies) involving 32,972 participants met our inclusion criteria. None of these studies assessed the effect of audit on quality from the women/mothers' perspective and none of the studies compared the effectiveness of different types of feedback. Ninety-five percentage (18/19) of studies showed significant improvement in at least one standard measured. Criterion-based audit has been used in obstetrics to improve quality from one dimension, namely the midwives/doctors' perspective. Midwives/doctors should consider the use of audit to improve quality of care from the mothers' view. There is need for well designed RCTs to assess the effectiveness of different types of feedback in criterion-based audit. PMID- 19557554 TI - Influence of highly active antiretroviral treatment (HAART) on risk factors for vertical HIV transmission. AB - OBJECTIVE: To analyze the influence of highly active antiretroviral treatment (HAART) on risk factors for perinatal transmission of human immunodeficiency virus (HIV). DESIGN: A prospective cohort study was performed between HIV pregnant women under HAART therapy and without treatment. SETTING: The maternity hospital 'La Fe' in Valencia, Spain. POPULATION OR SAMPLE: Five hundred HIV positive pregnant women. METHOD: Known maternal and obstetrical perinatal risk factors were analyzed by univariate and multivariate methods (logistic regression). The influence of HAART on the risk factors was evaluated independently to determine whether there was a modulation in perinatal HIV transmission. MAIN OUTCOME MEASURES: Known perinatal risk factors were found not to have any significant influence on perinatal HIV transmission in women under HAART therapy. RESULTS: Vertical transmission risk decreased significantly from 18.2% without treatment to 8.6% with mono/dual therapy and 0.6% with HAART. A CD4+ cell count below 500 cell/microl, intrapartum use of invasive procedures, rupture of membranes >six hours, labor length >five hours, and birthweight were the significant risk factors associated to vertical HIV transmission and elective cesarean section. Antiretroviral treatment administered during delivery was a protective factor in HIV pregnant women before HAART therapy. CONCLUSIONS: HAART therapy reduces the influence of the perinatal risk factors on vertical HIV transmission. PMID- 19557556 TI - Minimally invasive aortic valve replacement reduces atelectasis in cardiac intensive care. AB - OBJECTIVE: Respiratory failure is a major complication after cardiac surgery. The purpose was to evaluate the impact of minimally invasive aortic valve replacement (mini AVR) on the occurrence of left lower lobe atelectasis (LLLA) in the cardiac intensive care unit (ICU). PATIENTS AND METHODS: 98 patients were scheduled to undergo mini AVR. 14 of these patients were converted to a full sternotomy due to technical problems. These patients were compared to a group of 50 patients having planned AVR through a full sternotomy. The incidence of LLLA was evaluated on the first postoperative chest X-ray in the cardiac ICU. RESULTS: In the group having completed mini AVR 20/84 (24%) had a partial LLLA while in the group having extension to a full sternotomy 9/14 (64%) had LLLA lobe (P<0.005). In the group of 50 patients who had AVR through a full sternotomy, 27 patients (54%) had LLLA in the ICU which is also significantly higher (P<0.008) than the percentage of atelectasis in the mini AVR group. CONCLUSIONS: Patients who had mini AVR had a significantly lower incidence of LLLA in the cardiac ICU than patients who had AVR through a full sternotomy. PMID- 19557557 TI - Severe ataxia with neuropathy in hereditary gelsolin amyloidosis. AB - Hereditary gelsolin amyloidosis (AGel amyloidosis) is a systemic disorder caused by a G654A or G654T gelsolin mutation, reported from Europe, North America, and Japan. Principal clinical signs are corneal lattice dystrophy, cutis laxa and cranial neuropathy, often deleterious at advanced age. Peripheral neuropathy, if present, is usually mild. We report a 78- year old male Finnish patient who presented with ataxia and mainly sensory peripheral polyneuropathy (PNP) signs, causing severe disability and ambulation loss. Electrophysiological studies showed severe generalized chronic mainly axonal sensorimotor PNP with facial paralysis. In magnetic resonance imaging proximal lower limb and axial muscle atrophy with fatty degeneration as well as moderate spinal cord atrophy were seen. A G654A gelsolin mutation was demonstrated but no other possible causes of his disability were found. At age 79 he became bedridden and died of pulmonary embolism. Neuropathological examination revealed marked gelsolin amyloid deposition at vascular and connective tissue sites along the entire length of the peripheral nerves extending to the spinal nerve roots, associated with severe degeneration of nerve fibers and posterior columns. Our report shows that advanced AGel amyloidosis due to degeneration of central and distal sensory nerve projections results in deleterious ataxia with fatal outcome. Severe posterior column atrophy may reflect radicular AGel deposition, although even altered gelsolin - actin interactions in neural cells possibly contribute to neurodegeneration with successive ataxia in carriers of a G654A gelsolin mutation. PMID- 19557558 TI - Can a one-weekend group therapy reduce fear of blushing? Results of an open trial. AB - Fear of blushing is a specific syndrome generally subsumed under the diagnostic category of social anxiety disorder (SAD). This study aims at gathering preliminary data about an intensive weekend intervention specifically designed for individuals with fear of blushing as the predominant complaint. Treatment consisted of a combination of attention training and behavioral therapy. Thirty one blushing-fearful individuals meeting the criteria for SAD following the Diagnostic and Statistical Manual of Mental Disorders IV (DSM-IV) were treated in three groups. The study was conducted as an open trial. Full assessments were performed before treatment, six weeks after treatment, and at six-month follow up. Only fear of blushing, the main outcome criterion, was assessed immediately before and after the treatment weekend. The intensive therapy program was well accepted. Fear of blushing and SAD were significantly reduced and reductions remained stable. At follow-up, nearly two-thirds of the participants achieved significant changes in fear of blushing. Despite the preliminary nature of this study, the condensed format of weekend therapy for treating fear of blushing calls for further investigation. PMID- 19557559 TI - Fuchsine or magenta: the second most famous aniline dye. A short memoir on the 150th anniversary of the first commercial production of this well known dye. AB - During the mid-nineteenth century, it was learned that the distillation of coal tar yielded a mixture of benzene and toluene that could be used for the manufacture of "anilines." Oxidation with dichromate led to the first synthetic aniline dye, mauveine. The second aniline dye, a crimson red color, now is named fuchsine or magenta. This dye was prepared using the same starting material, but different oxidants, e.g., tin chloride, mercury nitrate, arsenic acid, and nitrobenzene. Unlike mauveine, which is now a chemical curiosity, fuchsine is still in use as a biological stain, especially in Schiff's reagent for detecting aldehydes, industrially as a dye in coloring various materials from textile fibers to ball point pen inks, analytically as a visualization agent for thin layer chromatography, and as an antifungal agent. PMID- 19557561 TI - False localizing signs in traumatic brain injury. AB - BACKGROUND: Hemiparesis ipsilateral to a mass-occupying lesion can be due to Kernohan-Woltman Notch Phenomenon (KWNP). This syndrome implies a false localizing sign because clinical findings lead the examiner to an incorrect neuroanatomical diagnosis. The contralateral crus cerebri (pyramidal tract) is pressed against the tentorial incisum and a resultant hemiparesis is found on the same side of the lesion. REVIEW: A detailed literature search of false-localizing signs is presented. CONCLUSIONS: Not infrequently, patients presenting to a physiatrist may have incomplete records. The existence of false localizing signs may point the physician towards the wrong underlying pathology. PMID- 19557563 TI - Occurrence of epilepsy during the first 10 years after traumatic brain injury acquired in childhood up to the age of 18 years in the south western Swedish population-based series. AB - BACKGROUND: The risk of seizures is increased after a traumatic brain injury (TBI), but the impact and duration of this increased risk is not well characterized in children. OBJECTIVE: To identify post-traumatic epilepsy (PTE) and post-concussion symptoms 10 years after a TBI during childhood. RESEARCH DESIGN: The study is a population-based retrospective follow-up study. PROCEDURE: Ten years after brain injury all 165 survivors, who as children (<18 years) in 1987-1991 as residents in the south western Swedish health care region had had a TBI, were invited to participate in a follow-up. A questionnaire regarding medical conditions and medication was filled out by the patients themselves or their parents as was a 21-item questionnaire (PCSQ) regarding post-concussion symptoms. Of the surviving 165 individuals, 109 participated (67%). RESULTS: Eight of 109 developed immediate seizures. During the follow-up period 12/109 had developed active epilepsy. Of these 12, five had had immediate seizures. The incidence of developing PTE within 10 years after a TBI was thus in this series 11%. The relative risk to develop late onset post-traumatic epilepsy (> or =1 week after injury) for those who had had immediate seizures was 9.018 (p = 0.0003, 95% CI = 3.69-22.05). CONCLUSIONS: TBI is a relatively rare cause of epilepsy in childhood, although immediate seizures are associated with an increased risk of developing post-traumatic epilepsy. PMID- 19557562 TI - Costs, mortality likelihood and outcomes of hospitalized US children with traumatic brain injuries. AB - PRIMARY OBJECTIVE: To examine the hospitalization costs and discharge outcomes of US children with TBI and to evaluate a severity measure, the predictive mortality likelihood level. RESEARCH DESIGN: Data from the 2006 Healthcare Cost and Utilization Project Kids' Inpatient Database (KID) were used to report the national estimates and characteristics of TBI-associated hospitalizations among US children < or =20 years of age. The percentage of children with TBI caused by motor vehicle crashes (MVC) and falls was calculated according to the predictive mortality likelihood levels (PMLL), death in hospital and discharge into long term rehabilitation facilities. Associations with the PMLL, discharge outcomes and average hospital charges were examined. RESULTS: In 2006, there were an estimated 58 900 TBI-associated hospitalizations among US children, accounting for $2.56 billion in hospital charges. MVCs caused 38.9% and falls caused 21.2% of TBI hospitalizations. The PMLL was strongly associated with TBI type, length of hospital stay, hospital charges and discharge disposition. About 4% of children with fall or MVC related TBIs died in hospital and 9% were discharged into long-term facilities. CONCLUSION: The PMLL may provide a useful tool to assess characteristics and treatment outcomes of hospitalized TBI children, but more research is still needed. PMID- 19557564 TI - Quality of life in children with acquired brain injury: parent perspectives 1-5 years after injury. AB - PRIMARY OBJECTIVE: To obtain parental ratings of children's quality of life, cognitive, emotional and behavioural functioning, as well as ratings of service provision, following traumatic brain injury (TBI). RESEARCH DESIGN: A retrospective, cross-sectional study. METHODS AND PROCEDURES: Parents of 47 children with mild or moderate-severe TBI completed standardized questionnaires evaluating quality of life (PedsQL 4.0) and cognitive, emotional and behavioural functioning (Strengths and Difficulties Questionnaire). Data collected was compared with published normative data for these scales. Views regarding parental experiences of care and their ratings of service provision were also obtained. RESULTS: Quality of life was significantly lower in 13-times as many children with TBI than expected from the normative population. Parents reported that more than 43% of children with TBI had cognitive, emotional and behavioural difficulties that impacted on their daily life. Whilst high levels of social deprivation were found, this did not fully explain the significantly raised levels of difficulties. Another factor associated with this poor outcome was the absence of systematic, routine follow-up or intervention. CONCLUSIONS: Parents frequently reported poor quality of life and cognitive, emotional and behavioural problems in their children following TBI. These preliminary findings indicate that children, after TBI, are at risk of developing persistent clinical problems and require follow-up beyond the acute period of their recovery. PMID- 19557565 TI - Anti-spastic effects of the direct application of vibratory stimuli to the spastic muscles of hemiplegic limbs in post-stroke patients. AB - OBJECTIVE: To investigate whether the direct application of vibratory stimuli inhibits spasticity and improves motor function in the hemiplegic upper limbs of post-stroke patients. DESIGN: Prospective pilot study. SETTING: University hospital rehabilitation centre. SUBJECTS: Fourteen post-stroke patients (mean age = 57.3 years; SD = 19.1 years). INTERVENTIONS: A hand and forearm stimulation device and an upper-arm stimulation device, consisting of vibrators, a wooden frame and a cloth strap, applied to the upper limbs of subjects. MAIN MEASURES: The modified Ashworth scale (MAS) score, F-wave parameters and motor-function parameters (finger tapping, active range of motion and the simple test for evaluating hand function). RESULTS: Subjects showed significant and potentially durable improvements in MAS score (p < 0.01), F-wave parameters (p < 0.01) and motor-function parameters (p < 0.05). The MAS score, F-wave parameters and motor function parameters dropped below the baseline values after vibratory stimulation. The MAS score and F-wave parameters remained significantly below the baseline 30 minutes after stimulation. CONCLUSIONS: The direct application of vibratory stimuli is an effective non-pharmacological anti-spastic treatment that could facilitate stroke rehabilitation. These results provide good evidence of potential short-term benefits of anti-spastic vibratory therapy in post-stroke patients in terms of decreased muscle tonus and improved motor function. PMID- 19557566 TI - Efficacy of therapeutic ultrasound and infrared in the management of muscle spasticity. AB - PRIMARY OBJECTIVE: This study was designed to compare the short-term effects of infrared (IR) and therapeutic ultrasound (US) on clinical and electrophysiological measures of spasticity and range of motion (ROM) in hemiplegic patients with plantar flexor muscle spasticity. RESEARCH DESIGN: A cross-sectional, single centre trial. INTERVENTIONS: Ultrasound and Infrared were used. METHODS AND PROCEDURES: Twenty-one patients (13 males and eight females) aged 27-78 years (mean +/- SD: 60.4 +/- 13.2) were randomly assigned into either the US or IR treatment groups. The IR and US were delivered for 20 and 10 minutes, respectively. The H-reflex, Ashworth scale (AS) and ROM were collected before, immediately after and 15 minutes after the end of the treatment. MAIN OUTCOMES AND RESULTS: Both groups were statistically similar on all variables at the beginning of the study. Results indicated that the dependent variables were not significantly different between groups at the end of study. The use of IR and US was not associated with significant reductions of H-reflex and Ashworth scores. A significant main effect for time on active and passive ROM was obtained. CONCLUSIONS: It was concluded neither IR nor US reduced electrophysiological or clinical measures of spasticity in this sample of patients. PMID- 19557567 TI - The impact of hypopituitarism on function and performance in subjects with recent history of traumatic brain injury and aneurysmal subarachnoid haemorrhage. AB - PRIMARY OBJECTIVE: To correlate deficient pituitary function with life satisfaction and functional performance in subjects with a recent history of traumatic brain injury (TBI) and subarachnoid haemorrhage (SAH). RESEARCH DESIGN: Cross-sectional study. METHODS AND PROCEDURES: Eighteen subjects with TBI and 16 subjects with SAH underwent pituitary hormonal and functional assessments 5-12 months following the event. Adrenal reserve was assessed with a 1 mcg cosyntropin stimulation test and growth hormone deficiency (GHD) was diagnosed by insufficient GH response to GHRH-Arginine stimulation. Assessments of life satisfaction and performance-function included the Satisfaction with Life Scale (SWLS), Craig Handicap Assessment and Reporting Technique (CHART) and the Mayo Portland Adaptability Inventory-4 (MPAI-4). RESULTS: Hypopituitarism was present in 20 (58.8%) subjects, including 50% with adrenal insufficiency. Hypothyroidism correlated with worse performance on SWLS and CHART measures. GHD was associated with poorer performance on CHART and MPAI-4 scale. CONCLUSIONS: In this series of subjects with history of TBI and SAH, hypothyroidism and GHD were associated with diminished life satisfaction and performance-function on multiple assessments. Further studies are necessary to determine the appropriate testing of adrenal reserve in this population and to determine the benefit of pituitary hormone replacement therapy on function following brain injury. PMID- 19557568 TI - The impact of alexithymia on somatization after traumatic brain injury. AB - INTRODUCTION: High rates of alexithymia have been reported following traumatic brain injury (TBI). Difficulty modulating emotional states has been shown to increase the risk of affective distress and the tendency to express this distress in the form of physical symptoms. The current study therefore examined relationships between alexithymia, affective distress and somatization in a TBI sample. METHOD: Eighty-three patients with TBI completed the Toronto Alexithymia Scale (TAS-20) and the Symptom Checklist (SCL-90-R). RESULTS: Alexithymic individuals reported higher ratings of anxiety, low mood and somatic symptoms. Alexithymia accounted for a significant amount of variance in anxiety, depression and somatization ratings. Scores on sub-scale 1 of the TAS-20 (difficulty identifying feelings) made a significant unique contribution to explaining somatization ratings after controlling for the influence of anxiety and depression ratings. CONCLUSION: Alexithymia after TBI increases the risk of affective disturbance and somatization. It needs to be identified at an early stage to direct rehabilitation interventions and improve prospects for psychosocial outcome. PMID- 19557569 TI - Use of the Personality Assessment Inventory (PAI) in individuals with traumatic brain injury. AB - PRIMARY OBJECTIVE: To evaluate the extent to which the Personality Assessment Inventory (PAI) is confounded by symptoms that are transdiagnostic between psychopathology and neurological sequelae. METHODS: Sixty-one adults with moderate-to-severe traumatic brain injury (TBI) completed the PAI over the first year post-injury. Items that discriminated brain-injured individuals from a normative sample were identified using effect size analyses and were then subjected to principal components analysis (PCA) with varimax rotation. To explore whether the items identified in the PCA may be considered transdiagnostic in nature, an expert rating task and correlations with objective outcome measures were employed. RESULTS: Effect sizes analyses identified 21 items that discriminated adults with TBI from the normative sample. Eight items associated with component 1 of the PCA were considered to be transdiagnostic. These items reflected health concerns and thinking problems from the Somatic Complaints, Depression and Schizophrenia scales. Items belonging to the other components reflected behaviours that are commonly associated with TBI, but not considered transdiagnostic. CONCLUSION: Using a comprehensive and multi-modal approach, results demonstrated good convergent validity for a small sub-set of items as being transdiagnostic. Overall, the findings support the PAI as a useful measure of psychiatric and emotional disturbances among persons with TBI. PMID- 19557571 TI - Comparative evaluation of corpus callosum DTI metrics in acute mild and moderate traumatic brain injury: its correlation with neuropsychometric tests. AB - PRIMARY OBJECTIVE: To look for differences in vulnerability of corpus callosum (CC) in patients of mild and moderate traumatic brain injury (TBI) in the acute stage using quantitative diffusion tensor imaging (DTI) and to correlate these with neuropsychometric tests (NPT) done at 6 months post-injury. RESEARCH DESIGN, METHODS AND PROCEDURES: Conventional MRI, DTI and NPT were performed on 83 patients (moderate TBI, n = 57; mild TBI, n = 26) within 5-14 days after TBI. Thirty-three age- and sex-matched healthy controls were also included for comparison. RESULTS: Significantly decreased fractional anisotropy (FA) in genu and splenium; significantly increased radial diffusivity (RD) values in genu, midbody and splenium with significant increase in mean diffusivity (MD) and a decrease in axial diffusivity (AD) only in genu, respectively, in patients with moderate TBI compared to healthy controls were observed. However, in moderate TBI, significantly decreased FA was found only in genu compared to mild TBI. Moderate TBI showed poor NPT scores compared to mild TBI, but this did not reach statistical significance. CONCLUSIONS: It is concluded that DTI abnormalities in the regions of CC were more in patients with moderate TBI compared to mild TBI and this was associated with relatively poor neuropsychological outcome 6 months post-injury. PMID- 19557570 TI - Assessment of thalamic perfusion in patients with mild traumatic brain injury by true FISP arterial spin labelling MR imaging at 3T. AB - OBJECTIVE: To assess cerebral blood flow (CBF) changes in patients with mild traumatic brain injury (MTBI) using an arterial spin labelling (ASL) perfusion MRI and to investigate the severity of neuropsychological functional impairment with respect to haemodynamic changes. MATERIALS AND METHODS: Twenty-one patients with MTBI and 20 healthy controls were studied at 3T MR. The median time since the onset of brain injury in patients was 24.6 months. Both patients and controls underwent a traditional consensus battery of neurocognitive tests. ASL was performed using true fast imaging with steady state precession and a flow sensitive alternating inversion recovery preparation. Regional CBF were measured in both deep and cortical gray matter as well as white matter at the level of basal ganglia. RESULTS: The mean regional CBF was significantly lower in patients with MTBI (45.9 +/- 9.8 ml/100 g min(-1)) as compared to normal controls (57.1 +/ 8.1 ml/100 g min(-1); p = 0.002) in both sides of thalamus. The decrease of thalamic CBF was significantly correlated with several neurocognitive measures including processing and response speed, memory/learning, verbal fluency and executive function in patients. CONCLUSIONS: Haemodynamic impairment can occur and persist in patients with MTBI, the extent of which is more severe in thalamic regions and correlate with neurocognitive dysfunction during the extended course of disease. PMID- 19557573 TI - A descriptive portrait of human assistance required by individuals with brain injury. AB - BACKGROUND: Human assistance is a counterweight to disabilities for people living with a traumatic brain injury (TBI). However, there is no clear description of the human assistance used by this population in relation with specific life habits (LH). OBJECTIVES: (1) to describe the proportion of LH performed with human assistance; (2) to explore the characteristics of TBI persons with greater needs for human assistance; (3) to clarify the categories of LH for which persons with TBI need human assistance; and (4) to determine the relationship between the human helper and the person with TBI across different residential settings. METHOD: One hundred and thirty-six individuals with moderate or severe TBI were interviewed using the LIFE-H. RESULTS: Human assistance is used to perform one out of three LH. A greater need for human assistance was associated with the number of impairments, motor limitation to the upper limbs, hemiplegia and receiving public insurance. Human assistance was used more often to perform LH pertaining to social roles than those pertaining to daily living. Close relatives were the most frequent providers of human assistance regardless of the residential setting. CONCLUSION: Given the importance of human assistance in TBI, it is essential to support human helpers during and after rehabilitation. PMID- 19557572 TI - A home programme for patients with cognitive dysfunction: a pilot study. AB - OBJECTIVE: To evaluate the effectiveness of a 12-week individualized home programme of rehabilitation for patients with cognitive impairment. METHODS: Six patients with cognitive dysfunction, after haemorrhagic stroke, participated in this study. A programme was carried out in the home environment that consisted of four mixed training programmes: cognitive remediation therapy, story retelling, cognitive enhancing games and aerobic exercise. The patients performed the home programme for 2 hours a day, 7 days a week, for 12 weeks. The main outcomes were measured using the MMSE, Neurobehavioural Cognitive Status Examination (NCSE), domain-specific computerized neuropsychological test for attention, memory and executive function, the Lowenstein Occupational Therapist Cognitive Assessment (LOTCA), the Modified Barthel Index (MBI) and the Seoul-Instrumental Activity of Daily Living (S-IADL). RESULTS: After completion of a 12-week home programme, the patients' S-IADL scores improved significantly (p < 0.05). The patients' MMSE and NCSE scores improved marginally, with mean score changes of 4.2 (p = 0.058), 4.7 (p = 0.078) and 6.8 (p = 0.068) points, respectively. However, the patients' domain-specific cognitive test and LOTCA scores did not significantly change. CONCLUSION: The results of this study showed that an individualized home programme improved IADL performance in patients with cognitive dysfunction and may have a beneficial effect on cognition, as assessed by general cognitive measures. PMID- 19557574 TI - Computerized tracking to train dexterity after cerebellar tumour: a single-case experimental study. AB - PRIMARY OBJECTIVE: In a child with pure ataxia after tumour removal, does dexterity training at the right elbow improve dexterity at the right elbow? Does improvement in dexterity transfer to improvement in upper limb activity? RESEARCH DESIGN: Single-case experimental study with 2-week baseline, 2-week intervention and 2-week follow-up phases and assessment by a blinded assessor. METHODS AND PROCEDURES: A 5-year old child 3 years after surgical resection of a low-grade cerebellar tumour underwent to dexterity training. Dexterity was measured using a Finger-to-Nose Test and upper limb activity was measured using the 9-Hole Peg Test. Measures were collected every 2 or 3 days over the 6-week period of the study. RESULTS: Finger-to-Nose Test showed visible but not statistical improvement between baseline and intervention phases in terms of smoothness of movement. On withdrawal of the intervention, visual analysis showed that scores remained at the same level. The 9-Hole Peg Test showed visible but not statistical improvement in terms of speed and smoothness of movement. On withdrawal of the intervention, visual analysis showed that improvement in speed was maintained during follow-up, but smoothness decreased. CONCLUSIONS: There is some evidence that dexterity can be trained and that improvement carries over to activity. PMID- 19557575 TI - FOS proliferating network construction in early colorectal cancer (CRC) based on integrative significant function cluster and inferring analysis. AB - The aim is to setup single distinguished molecular network. We constructed FOS proliferating network from 22 colorectal samples of the same GEO dataset by GRNInfer tool and DAVID based on linear programming and a decomposition procedure with integrated Kappa statistics and fuzzy heuristic clustering. In the control, we found no proliferating subnetwork. In CRC, we identified one FOS proliferating module (SFRP2, ADAMTS1, SYNPO2, VIP, ADAM33 inhibition to FOS and MGP, FOSB activation to FOS. FOS activation to IGFBP5, LGI1, GAS1 and FOS inhibition to VIP). These results may be useful for developing novel prognostic markers and therapeutic targets in CRC. PMID- 19557576 TI - Insights into apoptosis mechanisms induced by DNA-damaging agents in Burkitt's lymphoma cells. AB - p53 protein induces cell cycle arrest, DNA repair, or apoptosis of damaged cells. Loss of wild-type p53 (wt-p53) function was shown to be associated with upregulation of survivin and resistance to therapy. Here we investigated the effects of DNA-damage agents in inducing apoptosis and cell cycle arrest, and modulation of survivin levels in two Burkitt's lymphoma (BL) cell lines with different p53 mutations. Our results showed that BL cell lines have variable response to DNA-damaging agents that cannot be correlated exclusively with p53 mutation or survivin expression suggesting that p53-independent transactivation may play a role in apoptosis induced by DNA-damaging agents. PMID- 19557578 TI - Specific reduction of fas-associated protein with death domain (FADD) in clear cell renal cell carcinoma. AB - Fas-associated protein with death domain (FADD) plays a major role in the execution of apoptosis. Attenuation of apoptosis is a hallmark of cancer. Through systemic examination of FADD in renal cell carcinoma (RCC) and adjacent nontumor kidney tissues from 85 patients, we demonstrated a significant reduction of FADD in clear cell RCC (ccRCC) compared to the respective nontumor kidney tissues. In human kidney, FADD is expressed in both the proximal and distal tubules. As ccRCC originates from the proximal tubular epithelium, reduction of FADD in ccRCC indicates that FADD-mediated apoptosis may inhibit ccRCC tumorigenesis. PMID- 19557579 TI - Comparative effectiveness research in oncology: the need for clarity, transparency and vision. PMID- 19557582 TI - Individual models for virtual bone drilling in mastoid surgery. AB - Segmented training cases for virtual simulation of bone-drilling interventions in middle ear surgery have proven to be helpful in learning about surgical anatomy of the temporal bone. The anatomy of the mastoid shows a high degree of variability, however, and the aim of this study was to evaluate whether individual virtual models could be created within an affordable timeframe, and to what extend they reflected natural individual anatomy during virtual mastoid surgery. Automatic segmentation schemes were used, and these reduced the time required to create individual models on the basis of DICOM CT scans to less than 5 minutes. Models based on CT data with a slice distance of 0.4 mm or better were found to provide excellent handling, an acceptable depiction of mastoidal organs, and a helpful impression of the individual surgical situation. Although landmarks are still more easily detected in real mastoids, virtual drilling of individual models makes the 3D estimation of specific anatomy more effective than estimations based on interpretation of CT scans alone. PMID- 19557580 TI - The prognostic impact of bone marrow micrometastases in women with breast cancer. AB - Metastasis is the overwhelming cause of mortality in patients with breast cancer and our understanding of its cellular and molecular determinants is limited. Among cellular determinants, disseminated (DTC) tumor cells undoubtedly are key players in the metastatic cascade. In the past, several models have been constructed to explain the presence of individual tumor cells in secondary organs and their influence on the subsequent course of the disease. According to most recent transcriptome and genome analyses, DTCs are viewed as rare and much earlier indicators of metastasis than generally assumed from the typical course of breast cancer. Despite the observation that the numerous genetic alterations in such cells are rarely identical or even similar, characterization of the long interval between dissemination and clinically manifested metastases, the resistance to chemotherapy and significant effect on disease progression despite the low abundance in secondary organs, support the idea that some of these cells might be progenitor cells with self-renewing properties that give rise to most of the tumor mass that is dealt with clinically. Here, we review evidence from translational research and data from observational studies on DTCs to elucidate their potential impact for both future clinical trial design and, in the long run, decision-making in our daily patient management. PMID- 19557583 TI - Multi-factorial analysis of time efficiency in total knee arthroplasty. AB - The objective of this retrospective review was to determine whether time efficiency could be gained by optimizing the navigation protocol based on a surgeon's specific technique and work flow. Three groups of 30 consecutive patients operated on by the same surgeon were studied. The groups were from three distinct periods between 2002 and 2008. The first group consisted of patients in which no navigation was performed (the control group); the second group consisted of patients in which navigation was performed using a standard protocol; and the third group consisted of patients in which navigation was performed using a customized protocol that eliminated certain steps. Statistical analysis considered analysis of variance for covariates of total time in the operating room, duration of the procedure, and tourniquet time. Chi-square analysis considered categorical variables of age, gender, body mass index (BMI), Knee Society score, and patellar resurfacing against the surgical group. Multiple linear regression analysis evaluated surgical time adjusted for preoperative deformity, BMI, and patellar resurfacing. In the adjusted model, the customized navigation protocol significantly reduced the surgical time by 10 minutes compared to the non-customized navigation. Not resurfacing the patella significantly reduced the surgical time by 9 minutes. Variables of age, gender, BMI, preoperative deformity and Knee Society score were not related to differences in operating time. Time efficiency may be gained with the use of improved computer navigation protocols and patellar non-resurfacing. PMID- 19557584 TI - Changes in sexual risk taking with antiretroviral treatment: influence of context and gender norms in Mombasa, Kenya. AB - In-depth interviews were conducted with 23 sexually-active adults receiving antiretroviral treatment (ART) in Mombasa Kenya to understand changes in sexual behaviour after treatment initiation and factors influencing condom use. Advanced HIV disease had previously led to marked decreases in sexual desire and function. After HIV testing, numbers of partners reduced and monogamous relationships began to predominate. Receipt of ART strengthened these changes, while improving sexual health. However, concurrent sexual partnerships continue within polygamous marriage and unprotected sex occurs with regular partners, even those who are HIV negative. Those who used condoms inconsistently prior to ART often remained inconsistent users thereafter. While disclosure of HIV status appeared to support condom use, this does not always predict protected sex. In addition to classic perceptions about condom's effect on intimacy and trust, traditional gender roles, misconceptions about potential harm from condoms and fertility desires hinder condom use. PMID- 19557585 TI - Quality of offspring? Socio-cultural factors, pre-natal testing and reproductive decision-making in Japan. AB - Japan is one of the few countries to have enacted legislation on eugenics. Consequently, there has been active public debate about the practice of selective abortion for over 35 years. Furthermore, discrimination against disabilities is deep-rooted in Japanese society and the quality of offspring is a common concern. Given this background, the obvious conclusion might be that couples in Japan would have no hesitation in using reproductive technologies to ensure the best possible chance of giving birth to higher quality offspring. Yet, research indicates that when it comes to decision-making in the course of a pregnancy, not all individuals choose testing or termination, even when prenatal diagnosis indicates the presence of congenital conditions. Other factors play a role in reproductive decision-making, including age at time of pregnancy and reproductive history including infertility treatment. Against this background, this paper analyses accounts of five pregnancies - two of which resulted in termination and three which went to full term despite receiving test results showing possible birth defects - with the aim of identifying factors influencing whether or not to terminate a pregnancy. PMID- 19557586 TI - GNAS haploinsufficiency leads to subcutaneous tumor formation with collagen and elastin deposition and calcification. AB - INTRODUCTION: The heterotrimeric G protein alpha-subunit G(s)alpha links receptors to stimulation of cAMP/protein kinase A signaling, which inhibits skin fibroblast proliferation and collagen synthesis. We now describe the development of fibrous tumors in mice with heterozygous disruption of the Gnas gene, which encodes G(s)alpha and other gene products. METHODS AND RESULTS: Disruption of Gnas exon 2 on either the maternal or paternal allele (Gnas(E2-/+)) results in fibromas or angiofibromas on the ears, paws and tail beginning at 4 months of age. The tumors were composed of fibroblastic cell proliferation with collagen and elastin deposition and calcification, and seemed to be associated with mechanical skin damage. The presence of calcification was associated with greater amounts of matrix metalloproteinase-2, suggesting an association between calcium deposition and extracellular matrix degradation. Osteoblast-specific markers were absent, consistent with the calcification not being secondary to ossification. Molecular studies showed that the tumors were not associated with deletion of the wild-type allele, making it unlikely that these tumors resulted from homozygous loss of G(s)alpha. CONCLUSIONS: These findings provide in vivo evidence that G(s)alpha pathways inhibit fibroblast and endothelial proliferation and matrix deposition. PMID- 19557587 TI - Empty sella may be the final outcome in lymphocytic hypophysitis. AB - INTRODUCTION: Lymphocytic hypophysitis (LH) is an autoimmune disorder associated with the infiltration of the pituitary gland by lymphocytes leading to different degrees of hypopituitarism. Females are affected more frequently than males and the disease is usually associated with pregnancy or postpartum period. CASE: We present a case of LH who was first diagnosed with diabetes insipidus and hyperprolactinemia. In the follow-up, the patient developed growth hormone, gonadotropin, and thyroid stimulating hormone deficiency. The typical appearance of increased stalk thickness and diffuse homogenous contrast enhancement of pituitary on magnetic resonance imaging resulted in empty sella by time. CONCLUSION: The present case demonstrates the natural course of LH over a 13-year period in which the empty sella was the final outcome. PMID- 19557588 TI - Beneficial effects of sarpogrelate hydrochloride, a 5-HT2A receptor antagonist, supplemented with pioglitazone on diabetic model mice. AB - OBJECTIVE: Feeding behavior control and dietetics with consequent weight reduction can be the most efficacious and fundamental methods to normalize fasting blood glucose. However, pioglitazone treatment has been found to incrementally increase body weight. In this study, we investigated whether the combined application of a 5-HT(2A) receptor antagonist, sarpogrelate, with pioglitazone can provide a clinical benefit. METHODS: Diabetic male KK-A(y) mice were randomly assigned to four groups: those receiving 10 mg/kg/day pioglitazone treatment for 30 days (pioglitazone group, n = 7), those receiving 30 mg/kg/day sarpogrelate treatment for 30 days (sarpogrelate group, n = 7), those receiving both agents for 30 days (pioglitazone + sarpogrelate group, n = 7) and those receiving no treatment (control group, n = 7). RESULTS: Feed intake was lower in the pioglitazone + sarpogrelate group than in the pioglitazone group. Water intake was also significantly lower in the pioglitazone, sarpogrelate and pioglitazone + sarpogrelate groups than in the control group. Combined application (pioglitazone + sarpogrelate) resulted in a 176% increase in leptin concentration compared with vehicle control. Body weight was significantly higher in the pioglitazone group, and there was a trend toward a smaller increment in body weight in the pioglitazone + sarpogrelate group. Mean values, calculated by multiplying insulin concentration and nonfasting glucose concentration, were significantly lower in the pioglitazone + sarpogrelate group than in the control group. CONCLUSIONS: These results suggest that the combined application of sarpogrelate with pioglitazone provides therapeutic benefits not only in preventing adverse effects but also in the treatment of diabetes. PMID- 19557589 TI - Cytoplasm-to-nucleus shuttling of thyroid hormone receptor-beta1 (Trbeta1) is directed from a plasma membrane integrin receptor by thyroid hormone. AB - INTRODUCTION: In CV-1 cells, shuttling from cytoplasm to nucleus of the nuclear thyroid hormone receptor-beta1 (TRbeta1, TR) is shown in this report to be regulated by extracellular thyroid hormone at a hormone receptor on cell surface integrin alphav3. METHODS: The receptor was introduced into cells as a GFP-TR1 chimera and intracellular movement of the receptor was monitored by confocal microscopy of cells treated with L-thyroxine (T(4)). RESULTS AND DISCUSSION: TR GFP translocation in the presence of T(4) requires activation of extracellular regulated protein kinases 1/2 (ERK1/2). Inhibition of T(4)-binding to alphavbeta3 with anti-alphavbeta3 or Arg-Gly-Asp (RGD) peptide blocks T(4)-stimulated GFP-TR nuclear translocation, as do the hormone-binding inhibitor tetraiodothyroacetic acid (tetrac) and the ERK1/2 inhibitor, PD98059. TR1 is an ERK1/2 substrate. CONCLUSIONS: Via a nongenomic mechanism initiated at plasma membrane integrin v3, T(4)-activated ERK1/2 and TR1 move transiently in an immunoprecipitable complex to the nuclei of T(4)-treated cells. PMID- 19557593 TI - Obesity and insulin resistance increase plasma viscosity in young women with polycystic ovary syndrome. AB - OBJECTIVE: To investigate the plasma viscosity in young women with polycystic ovary syndrome (PCOS). DESIGN: Patients with PCOS and healthy controls were assessed for plasma viscosity. The acquired data were tested for association with hyperandrogenemia, obesity and insulin resistance (IR) in patients with PCOS. Plasma viscosity was determined by a viscometer Type 53610/I SCHOTT-Instruments, Mainz at 37 degrees C. PATIENTS: The study included 96 young women with PCOS and 72 healthy controls. Main outcome measures. Plasma viscosity and IR. RESULTS: Plasma viscosity was 1.243 +/- 0.67 mm(2)/s in the control group and 1.252 +/- 0.82 in women with PCOS (p = 0.416). Using multiple regression analysis, total protein (B = 0.348, p = 0.005), area under curve for insulin (B = 0.320, p = 0.011) and BMI (B = 0.315, p = 0.013) were proven to be significantly correlated to plasma viscosity. Plasma viscosity was significantly increased in women with PCOS with IR compared to matched for age and BMI PCOS women without IR (1.300 +/- 0.055 vs. 1.231 +/- 0.049 mm(2)/s) (p = 0.004). CONCLUSION: Young women with PCOS presented a plasma viscosity that was increased by obesity and IR. Therefore, clinical management of young overweight women with PCOS with IR should always include a serious reduction in body weight and the use of oral contraceptive treatment with cautious. PMID- 19557594 TI - The change in sex hormone binding globulin and the influence by gestational diabetes mellitus in fetal period. AB - OBJECTIVE: To investigate the influence of gestational diabetes mellitus (GDM) on the change of SHBG in fetus. METHOD: Forty-eight pregnant women with GDM and 86 women with normal pregnancy were included in the study. The following were measured in the serums of pregnant women, amniotic fluids, and umbilical cord serums: glucose, insulin, peptide-C, SHBG, and sex hormones. RESULTS: SHBGs in pregnant women's serums were, when compared with the control group: in male fetuses 308.06 +/- 55.64 vs. 445.21 +/- 50.07 (p < 0.01) and in female fetuses 312.38 +/- 56.61 vs. 451.05 +/- 52.87 (p < 0.01). When comparing the levels of SHBGs in amniotic fluids, inclusive of the control group, the following were in male fetuses 8.35 +/- 1.07 vs. 8.41 +/- 1.09 (p = NS) and in female fetuses 8.31 +/- 0.97 vs. 8.39 +/- 0.94 (p = NS). For the levels of SHBGs in umbilical cord serums and comparison to the control group were: in male fetuses 41.44 +/- 8.83 vs. 40.24 +/- 7.50 (p = NS) and in female fetuses 39.93 +/- 7.04 vs. 39.69 +/- 7.16 (p = NS). The concentration of SHBG in amniotic fluid had no significant relationship to glucose, dehydroepiandrosterone (DHEAS), estradiol, and total and free testosterone, but had an extremely negatively correlated to insulin and peptide-C (p < 0.01) in GDM group. CONCLUSION: Although the concentration of SHBG does not change in fetus when pregnant woman is complicated with GDM, it is already influenced by the fetal regulation on hyperinsulinemia. PMID- 19557595 TI - Protein profile of the luteal phase endometrium by tissue microarray assessment. AB - To investigate the luteal phase endometrial expression of leukemia inhibitor factor (LIF), insulin-like growth factor 1 (IGF-1), progesterone receptor (PR), claudin 4 (CLDN4), vascular-endothelial growth factor receptor 3 (VEGFR-3), bone morphogenetic protein 4 (BMP-4) and citokeratin 7 (CK-7), we obtained luteal phase endometrial samples from 52 women. Samples were dated and integrated using a tissue microarray (TMA). Samples were immunostained for LIF, IGF-1, PR, CLDN4, VEGFR-3, BMP-4 and CK-7. Frequencies of positive expressions at the early, mid and late luteal phases were compared by two proportions test. Concomitant expression of these proteins was assessed with Chi-square or Fischer's test. The frequency of LIF was positively correlated to the frequency of IGF-1 (r = 0.99; p < 0.05) and PR (r = 0.99; p < 0.05), and the correlation between IGF-1 and PR tended to be significant (r = 0.98; p < 0.1). The expression of PR was associated with the absence of CLDN4 (p < 0.001). Thus, expression of LIF, IGF-1 and PR are correlated during the luteal phase, and immunohistochemistry for these proteins might be used to assist in the assessment of endometrial maturation. In addition, the expression of CLDN4 and PR was not concomitant, warranting further investigation on the relationship of their endometrial expression. PMID- 19557590 TI - The effect of gonadectomy and estradiol on sensitivity to oxidative stress. AB - The sexual dimorphism of life span and caloric restriction effects in numerous species suggest that estradiol (E2) is protective against oxidative damage. The only direct test of E2's protective effect in mice against in vivo oxidative stress to date may have been confounded by E2's direct chemical action as an antioxidant because it was administered at very high dosages. Therefore, we have identified a low yet physiologically effective dose of E2. We then administered this dose using subcutaneous time-release pellets to ovariectomized mice. Two weeks after E2 pellet implantation, sham-operated, ovariectomized, and ovariectomized E2-supplemented female mice were injected with a lethal dose of paraquat and their survival was followed. It was observed that ovariectomy exacerbates paraquat-induced mortality and is rescued by E2 supplementation. An equivalent experiment was performed on sham-operated, orchidectomized, and E2 supplemented orchidectomized male mice. The survival of male mice was improved by orchidectomy, and E2 gave no further benefit. We interpret the results to mean that E2 is protective against oxidative stress through its regulatory role and that testosterone diminishes protection against oxidative stress. PMID- 19557598 TI - Building characteristics associated with moisture related problems in 8,918 Swedish dwellings. AB - Moisture problems in buildings have in a number of studies been shown to increase the risk for respiratory symptoms. The study Dampness in Buildings and Health (DBH) was initiated with the aim to identify health relevant exposures related to dampness in buildings. A questionnaire study about home environment with a focus on dampness problems and health was conducted in one county of Sweden (8,918 homes, response rate 79%). Building characteristics that were associated with one or more of the dampness indicators were for single-family houses, older houses, flat-roofed houses built in the 1960s and 1970s, houses with a concrete slab on the ground that were built before 1983. Moreover, tenancy and earlier renovation due to mould or moisture problems was strongly associated with dampness. A perception of dry air was associated with window-pane condensation, e.g. humid indoor air. PMID- 19557599 TI - History and development of radiation-protective agents. AB - PURPOSE: The search for ideal protective agents for use in a variety of radiation scenarios has continued for more than six decades. This review evaluates agents and procedures that have the potential to protect against acute and late effects of ionising radiation when administered either before or after radiation exposure. CONCLUSION: Over the years, extensive experimental studies of radiation protective agents have enhanced our knowledge of radiation physics, chemistry, and biology. However, translation of agents from animal testing to use in various scenarios, such as prophylactic adjuncts in radiotherapy or post-exposure treatments for potential victims of radiation accidents/incidents, has been slow. Nevertheless, a number of compounds are now available for use in a variety of radiation situations. These include agents approved by the U.S. Food and Drug Administration for use in reducing exposure to internal radionuclides (Prussian blue, calcium diethylenetriamene pentaacetate (DTPA) and zinc DTPA, potassium iodide) and amifostine for alleviating xerostomia associated with radiotherapy. Consensus groups have also recommended other therapies such as granulocyte colony stimulating factor for radiation-induced neutropenia. The variety of prophylactic and therapeutic agents in the research pipeline includes those that are naturally occurring with low toxicity, provide a long window of protection, protect normal tissue while sensitising tumours, or act via receptors and modulate biological processes such as induction of genes responsible for radioresistance. The search for agents that protect against acute and late effects of ionising radiation injury will undoubtedly continue into the future and influence other areas of radiation research. PMID- 19557600 TI - Milestones in normal tissue radiation biology over the past 50 years: from clonogenic cell survival to cytokine networks and back to stem cell recovery. AB - PURPOSE: To illustrate the progress in normal tissue radiation biology over the last five decades and its impact on radiotherapy. MATERIALS AND METHODS: Major milestones over the last 50 years and their consequences for radiation oncology are described: The identification of clonogenic cell survival and the (target) stem cell concept, the dissociation between early and late responding tissues with regard to dose fractionation and development of the linear-quadratic model, characterisation of the effect of overall treatment time, the definition of retreatment tolerance. Current knowledge of mechanisms of radiation pathogenesis is a basis for most recent approaches for amelioration of normal tissue effects. RESULTS: Advances in radiobiological research in normal tissues in the last 50 years have had a major impact on radiation oncology. This includes the linear quadratic model to adjust doses in altered fractionation protocols, and quantitation of repopulation processes to avoid toxicities in accelerated regimen. Based on new insights into the pathogenesis of normal tissue radiation effects, promising strategies for their modulation, e.g., with cytokines or by stem cell therapy, have been developed. CONCLUSIONS: Research on radiobiology with relevant in vivo models, and relevant treatment protocols is essential for the further progress in radiation oncology. PMID- 19557601 TI - Gamma-tocotrienol, a tocol antioxidant as a potent radioprotector. AB - PURPOSE: To assess the radioprotective potential of gamma-tocotrienol. MATERIALS AND METHODS: To optimise its dose and time regimen, gamma-tocotrienol (GT3) was injected subcutaneously (SC) at different doses into male CD2F1 mice [LD(50/30) (lethal radiation dose that results in the mortality of 50% mice in 30 days) radiation dose of 8.6 Gy with vehicle]. The mice were given 10.5, 11 and 11.5 Gy cobalt-60 radiation, and 30-day survival-protection was determined. Time optimisation was done by SC administration of GT3 at different intervals before irradiation. Dose reduction factor (DRF) was determined by probit analysis using mortality as the end point at six radiation doses. Protection from radiation induced pancytopenia was determined by enumerating peripheral blood cells from mice given GT3 and irradiated at 7 Gy. RESULTS: At an optimal dose of 200 mg/kg given SC 24 h before irradiation, GT3 had a DRF of 1.29. GT3 accelerated the recovery of total white blood cells, neutrophils, monocytes, platelets, and reticulocytes in irradiated mice, compared to vehicle-injected, irradiated controls. CONCLUSION: GT3 is a radioprotectant having a higher DRF than any other tocols. The protection it provides close to the gastro-intestinal range indicate that GT3 can be considered as an ideal radioprotectant meriting further drug development stages for the ultimate use in humans. PMID- 19557602 TI - The electric field is a sufficient physical determinant of the human magnetic sense. AB - PURPOSE: The onset and offset of weak low-frequency magnetic fields triggered evoked potentials in human subjects that could be detected using nonlinear analysis, but not by means of time averaging. Because the magnetic fields and their induced electric fields were both present in the brain, their respective role in producing the effect on brain activity could not be ascertained. We inquired whether a biophysical coupling mechanism involving only the electric field could explain the occurrence of the brain potentials. MATERIALS AND METHODS: An external electric field capable of producing a brain electric field comparable to that induced by the magnetic stimuli was identified by finite element analysis. The electroencephalogram from 23 subjects was measured from six scalp derivations in the presence and absence of the external electric field, and the presence of evoked potentials was assessed using nonlinear and linear analyses. RESULTS: Evoked potentials were observed in all but one subject (p < 0.05 in each subject); the potentials had the same latency, duration, and distribution of magnitudes as seen in the earlier studies, and were detectable only by means of nonlinear analysis. Using a realistic physical model of an ion channel, we showed that transduction of an electric field could be explained by assuming that the field exerted a force on glycocalyx molecules attached to a channel gate. CONCLUSION: The evoked potentials described here, as well as those observed previously in response to magnetic stimuli, were probably triggered by the induced electric field. PMID- 19557603 TI - Static magnetic field attenuates mortality rate of mice by increasing the production of IL-1 receptor antagonist. AB - PURPOSES: Disseminated intravascular coagulation (DIC) is a complex systemic thrombohemorrhagic disorder involving intravascular coagulation and hemorrhage. The aim of this study is to test whether static magnetic field (SMF) is effective in attenuating lipopolysaccharide (LPS)-induced DIC. MATERIALS AND METHODS: In vivo experiments were performed in this study using male BALB/cByJ mice. An intraperitoneal injection of 50 mg/kg LPS was shown to lead to approximately 50% mortality and this dose was used in subsequent experiments. To test the effects of SMF on the survival rate of LPS-induced animals, the mice were exposed to 0.25 T SMF for 2 h before LPS injection. In addition, the effect of a 2-h SMF treatment on the production of anti-inflammatory cytokines was evaluated. RESULTS: In the first set of experiments, we found that the survival rate was higher in the SMF-exposed group than in the sham-exposed group. The circulating platelet (PLT) counts in the SMF-exposed mice were significantly higher than in the unexposed animals. However, no significant changes in inflammatory cytokine, including tumour necrosis factor-alpha (TNF-alpha), interleukin-1alpha (IL 1alpha), interleukin-6 (IL-6) and monocyte chemotactic protein 1 (MCP-1), in plasma were found after SMF treatment. The results from the second experiment showed that the plasma levels of interleukin-1 receptor antagonist (IL-1ra) were higher in the SMF-exposed group than in the sham group. CONCLUSIONS: Exposure to an SMF increases the plasma levels of IL-1ra. This effect may inhibit the reduction in PLT in plasma, resulting in prevention in LPS induced DIC. PMID- 19557605 TI - Area assessment of psoriasis lesions for PASI scoring. AB - Psoriasis is a skin disorder which is caused by a genetic fault. Although there is no cure for psoriasis, there are many treatment modalities to help control the disease. To evaluate treatment efficacy, the current gold standard method, PASI (Psoriasis Area and Severity Index), is used to measure psoriasis severity by evaluating the area, erythema, scaliness and thickness of the plaques. However, the determination of PASI can be tedious and subjective. In this work, we develop a computer vision method that determines one of the PASI parameters, the lesion area. The method isolates healthy and healed skin areas from lesion areas by analysing the hue and chroma information in the CIE L*a*b* colour space. Centroids of healthy skin and psoriasis in the hue-chroma space are determined from selected sample. The Euclidean distance of all pixels from each centroid is calculated. Pixels are assigned to either healthy skin or psorasis lesion classes based on the minimum Euclidean distance. The study involves patients from different ethnic origins having three different skin tones. Results obtained show that the proposed method is able to determine lesion areas with accuracy higher than 90% for 28 out of 30 cases. PMID- 19557606 TI - Porcine skin as a model system for studies of adverse effects of narrow-band UVB pulses on human skin. AB - Ultraviolet (UV) radiation has been widely used in medicine, and in recent years there has been a growing interest in narrow-band UVB therapies, especially those employing pulses of the 308-nm line of XeCl excimer lasers. Comparative studies in several skin pathologies showed that narrow-band UVB was more effective than classical broad-band UVB radiation. Simultaneously, UVB is carcinogenic and there is a need for data to establish the risk associated with phototherapies involving irradiations of human skin with different doses of narrow- and broad-band UVA and/or UVB radiation. Relevant data are sparse predominantly due to a lack of suitable model systems for study of this phenomenon. Our comparative study of human and porcine skin responses to pulses of narrow-band UVB radiation demonstrated that for doses ranging from 5 to 10,000 mJ/cm(2) both skin types have similar susceptibility to UVB-induced breaking of nuclear DNA, indicating that pig skin might serve as good model for studies of sensitivity of human skin to UVB radiation. PMID- 19557607 TI - Percutaneous absorption of an insect repellent p-menthane-3,8-DIOL: a model for human dermal absorption. AB - p-Menthane-3,8-diol(38DIOL) was recently introduced as a natural topical insect repellent in the commercial product "OFF! Botanicals" lotion. The objective of this study was to provide an estimate of the potential for 38DIOL systemic absorption in humans. Carbon-14-labeled 38DIOL formulated in the lotion and in an ethanol solution was applied to excised pig skin in an in vitro flow-through test system predictive of skin absorption in humans. Twenty-four hours after application, radiolabel recovered from the dermis and receptor fluid was summed to determine percent absorption. At a dose of approximately 80 microg/cm(2) of 38DIOL in the lotion, a value of 3.5 +/- 0.8% of applied dose was obtained with pig skin. The corresponding value for 38DIOL in ethanol (90 microg/cm(2)) was not significantly different (3.0 +/- 1.2%). Most of the applied dose of 38DIOL was found to evaporate from pig skin (77 +/- 8% for the lotion and 87 +/- 1% for ethanol solution), thus limiting percutaneous absorption values. For reference purposes, the pig skin absorptions of piperonyl butoxide (PBO) at 100 microg/cm(2) in isopropanol, N,N-diethyl-m-toluamide (DEET) at 500 microg/cm(2) in ethanol, and neat isododecane at 650 microg/cm(2) (in order of increasing volatility) were 15 +/- 6%, 23 +/- 3%, and 0.09 +/- 0.05% of applied dose respectively. Isododecane was lost almost exclusively from the skin surface by evaporation. For additional reference, absorptions of PBO, DEET, and 38DIOL were found to be higher with excised rat skin. PMID- 19557608 TI - Bioavailability of tungsten and associated metals in calcareous soils in the vicinity of an ancient metalliferous mine in the Corbieres area, southwestern France. AB - The mobility and bioavailability of tungsten and associated metals are examined in calcareous soils and subsequent bioaccumulation by four species of plants is determined. Apparent bioavailability of metalliferous cations indicated by accepted monitoring methods and actual bioaccumulation is compared using regression analysis. Two soil extraction procedures were used without significant correlation between the methods at all stages, with the exception of copper and arsenic. More importantly, perhaps, the bioaccumulation by various tissues of Buxus sempervirens did not significantly correlate for the majority of target metals for each extraction procedure. Possible accumulation of toxic cations by a dying tree species was also examined. The availability of tungsten and associated metals in calcareous soils was compared with previous investigations on acidic soils, resulting in confirmation that tungsten in particular, in naturally occurring ores, is more readily mobilized under alkaline conditions. PMID- 19557609 TI - Air pollution and emergency room visits for arrhythmias: are there potentially sensitive groups? AB - Recent studies showed that air pollution is a risk factor for hospitalization for arrhythmias. However, there is limited evidence to suggest which subpopulations are at higher risk for arrhythmia development due to increased air pollutant exposure. This study was undertaken to examine the modifying effect of specific secondary diagnosis (including hypertension, diabetes, and congestive heart failure) on the relationship between frequency of emergency room (ER) visits for arrhythmias and ambient air pollutants concentrations. ER visits for arrhythmias and ambient air pollution data for Taipei were obtained for the period 2000-2006. The relative risk of ER visits was estimated using a case-crossover approach. Data showed an increased risk of ER visits for arrhythmias in relation to increased O(3) levels among individuals with a secondary diagnosis of hypertension and congestive heart failure. PMID- 19557610 TI - Long-term perchloroethylene exposure: a meta-analysis of neurobehavioral deficits in occupationally and residentially exposed groups. AB - The literature concerning the neurobehavioral and neurophysiological effects of long-term exposure to perchloroethylene (PERC) in humans was meta-analyzed to provide a quantitative review and synthesis in the form of dose-effect curves. The useable database from this literature comprised studies reporting effects of long-term exposure to PERC, effects that included slowed reaction times, cognitive deficits, impaired color vision, and reduced visual contrast sensitivity. For the meta-analyses, dose was defined as the product of the concentration inhaled PERC and the duration of exposure, expressed in unites of ppm-h/1000 (for numerical convenience). Dose-related results were highly variable across studies. Reports involving low exposure concentrations characteristic of nonoccupational exposures consistently produced effects of a magnitude that were comparable to those reported for higher concentration occupational studies. If this finding is reliable and general, studies of occupationally exposed persons may underestimate the magnitude of effects of PERC and other chemicals in the total population. Given the limited scope of the available data for PERC and its methodological and reporting problems (small sample sizes, testers were not blind to the subjects' exposure conditions, and the timing and location of testing were insufficiently documented), it seems important to test this conclusion with a well-documented study of two groups (occupational and nonoccupational exposure) in which subjects are evaluated in randomized order, using the same procedures and with the testers kept blind to the status of the subjects. PMID- 19557611 TI - Dermal permeation of biocides and aromatic chemicals in three generic formulations of metalworking fluids. AB - Metalworking fluids (MWF) are complex mixtures consisting of a variety of components and additives. A lack of scientific data exists regarding the dermal permeation of its components, particularly biocides. The aim of this study was to evaluate the dermal permeation of biocides and other aromatic chemicals in water and in three generic soluble oil, semi-synthetic, and synthetic MWF types in order to evaluate any differences in their permeation profiles. An in vitro flow through diffusion cell study was performed to determine dermal permeation. An infinite dose of different groups of chemicals (6 biocides and 29 aromatic chemicals) was applied to porcine skin, with perfusate samples being collected over an 8-h period. Perfusate samples were analyzed by gas chromatography/mass spectrometry (GC-MS) and ultra-performance liquid chromatography/mass spectroscopy (UPLC-MS), and permeability was calculated from the analysis of the permeated chemical concentration-time profile. In general, the permeation of chemicals was highest in aqueous solution, followed by synthetic, semi-synthetic, and soluble oil MWF. The absorption profiles of most of the chemicals including six biocides were statistically different among the synthetic and soluble oil MWF formulations, with reduced permeation occurring in oily formulations. Permeation of almost all chemicals was statistically different between aqueous and three MWF formulation types. Data from this study show that permeation of chemicals is higher in a generic synthetic MWF when compared to a soluble oil MWF. This indicates that a soluble oil MWF may be safer than a synthetic MWF in regard to dermal permeation of chemicals to allow for an increased potential of systemic toxicity. Therefore, one may conclude that a synthetic type of formulation has more potential to produce contact dermatitis and induce systemic toxicological effects. The dilution of these MWF formulations with water may increase dermal permeability of biocides, allowing for an enhanced risk for systemic toxicological effects and dermatitis potential. PMID- 19557612 TI - Comparing water, bovine milk, and indoor residual spraying as possible sources of DDT and pyrethroid residues in breast milk. AB - The presence of pollutants in human breast milk is of major concern, especially in malaria control areas where 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) is currently used as indoor residual spray (IRS). The levels of DDT and pyrethroids (PYR) were determined in breast milk, bovine milk, and drinking water from northern KwaZulu-Natal, South Africa. Both reference and exposed mothers used the same market food, but the DDT levels in the exposed mothers (mean SigmaDDT 10 microg/g milk fat [mf]) were much higher than for the reference mothers (mean SigmaDDT 1.3 microg/g milk fat). This difference in residue levels indicates uptake from IRS-applied DDT, most likely via air and skin contact, and excludes food as the main source of pollutants. DDT levels in bovine milk (mean SigmaDDT 0.15 microg/g mf) from the exposed area were less than levels in breast milk from the reference area, and lower than the 20 microg/L maximum residue limit (MRL) set by the Food and Agriculture Organization (FAO). Mean SigmaDDT in water was 0.0065 microg/L, much lower then the WHO limit of the sum of all metabolites in drinking water of 1 microg/L, and therefore highly unlikely to have contributed to any extent toward levels in breast milk. Permethrin in breast milk (mean 1.1-1.6 microg/g milk fat) was probably derived from home garden and indoor use, while the other PYR (cypermethrin and cyfluthrin) at lower concentrations were probably derived from food and agricultural exposure. It is postulated that a better understanding of the indoor dynamics of DDT and other insecticides, through a concept of Total Homestead Environment Approach (THEA), is crucial for investigating options of reducing human exposure and uptake under malaria control conditions. PMID- 19557613 TI - Factors affecting mercury and selenium levels in New Jersey flatfish: low risk to human consumers. AB - Some fish contain high levels of mercury (Hg), which could pose a risk to fish eaters themselves or their children. In making decisions about fish consumption, people must decide whether to eat fish, how much to eat, what species to eat, and what size fish to eat, as well as suitable (or unsuitable) locations, among other factors. Yet to make sound decisions, people need to know the levels of Hg in fish as a function of species, size, and location of capture. Levels of Hg and selenium (Se) were examined in three species of flatfish (fluke or summer flounder [Paralichthys dentatus], winter flounder [Pseudopleuronectes americanus], and windowpane [Scophthalmus aquosus]) from New Jersey as a function of species, fish size, season, and location. Flatfish were postulated to have low levels of Hg because they are low on the food chain and are bottom feeders, and data were generated to provide individuals with information on a species that might be safe to eat regularly. Although there were interspecific differences in Hg levels in the 3 species, total Hg levels averaged 0.18, 0.14, and 0.06 ppm (microg/g, wet weigh) in windowpane, fluke, and winter flounder, and selenium levels averaged 0.36, 0.35, and 0.25 ppm, respectively. For windowpane, 15% had Hg levels above 0.3 ppm, but no individual fish had Hg levels over 0.5 ppm. There were no significant seasonal differences in Hg levels, although Se was significantly higher in fluke in summer compared to spring. There were few geographical differences among New Jersey locations. Correlations between Hg and Se levels were low. Data, based on 464 fish samples, indicate that Hg levels are below various advisory levels and pose little risk to typical New Jersey fish consumers. A 70-kg person eating 1 meal (8 oz or 227 g) per week would not exceed the U.S. Environmental Protection Agency reference dose of 0.1 microg/kg body weight/d of methylmercury (MeHg). However, high-end fish eaters consuming several such meals per week may exceed recommended levels. PMID- 19557615 TI - The toxic effects of multiple persistent organic pollutant exposures on the post hatch immunity maturation of glaucous gulls. AB - This study tested whether the immune system of the glaucous gull (Larus hyperboreus) chicks became affected by existing environmental contaminants. An experimental group was given food that mimicked the natural contaminant mixture found in food from the North Atlantic marine environment, while the control group was given the equivalent of nearly clean food. All chicks were immunized with herpes virus (EHV), reovirus (REO), influenza virus (EIV), and tetanus toxoid (TET) in order to test their ability to respond to foreign specific antigens. At 8 wk, the experimental group had 3- to 13-fold higher concentrations of hexachlorobenzene (HCB), oxychlordane, p,p'-DDE, and total polychlorinated biphenyls (Sigma PCB) than did the control. The experimental group produced significantly lower antibody titer against EIV and had lower concentrations of immunoglobulin-G (IgG) and -M (IgM) in blood. Hematocrit percent and leukocyte numbers did not differ between the two groups. The ability of lymphocytes to proliferate in vitro was tested with three mitogens, phytohemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM), and three antigens, keyhole limpet hemocyanin (KLH), TET, and Mycobacterium avium subsp. paratuberculosis tuberculin purified protein derivative (PPD). The experimental group had a significantly higher peripheral blood lymphocyte response to PHA and to spleen lymphocytes in vitro stimulated with Con A and PCB congeners 99 or 153, while the Con A, PWM, KLH, TET, PPD, and Con A plus PCB-156 or -126 showed nonsignificant differences between groups. Data indicate that the combined effect of multiple persistent organic pollution exposures occurring naturally in the Arctic negatively affect the immune system of the glaucous gull chick. PMID- 19557616 TI - Acute and subchronic toxicity of arsenite and zinc to tadpoles of Rhinella arenarum both alone and in combination. AB - The current study evaluated acute and subchronic toxicity of arsenite (As(3+)) and zinc (Zn(2+)) to stage 25 tadpoles of Rhinella arenarum in both single and joint laboratory exposures. LC50 values obtained for As(3+) were elevated and remained within the range of 46 to 50 mg/L of As(3+) between 4 and 17 d of exposure. Growth of tadpoles was completely inhibited with 30 mg/L of As(3+), demonstrating the presence of ecologically relevant sublethal effects at concentrations lower than those resulting in lethality. With respect to Zn(2+), a 96-h LC50 value of 2.49 mg/L was calculated in soft water. Contrary to results obtained for As(3+), LC50 values of Zn(2+) gradually decreased with increasing exposure duration, from 2.49 mg/L at 96 h to 1.30 mg/L after 21 d. In joint exposures to both metals, the type of interaction observed between As(3+) and Zn(2+) was concentration dependent. Lethal effects of As(3+) were mitigated, unaffected, or potentiated by 0.01, 0.1, and 1-2 mg/L of Zn(2+), respectively. However, although 0.01 mg/L of Zn(2+) significantly reduced lethality of As(3+) exposed tadpoles, the same concentration of Zn(2+) did not help to reverse the stunt growth of these animals. Further studies need to examine which are the lowest concentrations As(3+) required to reduce growth and whether Zn(2+) serves to antagonize growth effects in this range of concentrations. PMID- 19557614 TI - Comparative covalent protein binding of 2,5-hexanedione and 3-acetyl-2,5 hexanedione in the rat. AB - 2,5-Hexanedione (HD) is the metabolite implicated in n-hexane neurotoxicity. This gamma-diketone reacts with protein lysine amines to form 2,5-dimethylpyrrole adducts. Pyrrole adduction of neurofilaments (NF) and/or other axonal proteins was proposed as a critical step in the neuropathy. While pyrrole adduction is widely accepted as necessary, subsequent pyrrole oxidation, which may result in protein cross-linking, was alternatively postulated as the critical mechanistic step. Previous studies have indicated that 3-acetyl-2,5-HD (AcHD), an analogue that forms pyrroles that do not oxidize, was not neurotoxic in rats. However, relative levels of pyrrole adduction of NF or other axonal proteins were not reported. In the present study, groups of 6 male Wistar rats were given saline, [1,6-(14)C]-HD (3 mmol/kg/d), or [5-(14)C]-AcHD (0.1 mmol/kg/d), i.p. for 21 d. HD- and AcHD-treated rats lost 10% and gained 14% body weight, respectively, compared to a 22% gain for control rats. At termination, HD- and AcHD-treated rats exhibited mean scores of 3.5 and 1.4, respectively, for hindlimb weakness (0 5 scale). Incorporation of radiolabel from HD was 27.8 +/- 3.9, 13.9 +/- 2.6, and 7.8 +/- 0.6 nmol/mg in plasma protein, purified globin, and axonal cytoskeletal proteins, respectively, compared to 0.6 +/- 0.1, 1.6 +/- 0.5, and 1.0 +/- 0.1 for AcHD. Binding of HD to the NF-L, -M, and -H subunit proteins from treated animals was 4-, 24-, and 13-fold higher, respectively, that that of AcHD, indicating differing stoichiometry and patterns of NF adduction for the two diketones. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of globin and NF proteins did not demonstrate protein cross-linking for either diketone at the dose levels and time period examined. These results indicate that that the lack of neurotoxicity previously reported for AcHD may reflect differences in adduct levels at critical axonal target sites rather than an inability to form cross-linking adducts. Based on these data, further studies are required to fully assess the neurotoxic potency of AcHD and other non-cross linking analogues as compared to HD. PMID- 19557617 TI - Urinary porphyrin excretion in children with mercury amalgam treatment: findings from the Casa Pia Children's Dental Amalgam Trial. AB - Increases in the urinary concentrations of pentacarboxyl- and coproporphyrins and the appearance of the atypical precoproporphyrin have been defined in relation to mercury (Hg) body burden in animal studies, and this change in the porphyrin excretion pattern has been described as a biomarker of occupational Hg exposure and toxicity in adult human subjects. In the present studies, urinary porphyrins were determined in relation to Hg exposure in children and adolescents, 8-18 yr of age, over the 7-yr course of a clinical trial designed to evaluate the neurobehavioral and renal effects of dental amalgam in children. Subjects were randomized to either dental amalgam or composite resin treatments. Urinary porphyrins and creatinine concentrations were measured at baseline and annually in all subjects. Results were evaluated using linear regression analysis. No significant differences between treatment groups (amalgam versus composite) were found when comparing all subjects for any of the porphyrins of interest. However, incipent amalgam treatment-specific increases were observed in the mean concentrations of penta-, precopro- and coproporphyrins especially when the analyses were restricted to younger subjects (8 to 9 yr old at baseline), and these increases were most apparent during yr 2 through 3 of follow-up, the period of highest mercury exposure from amalgam treatment. Based on the mean number of amalgam fillings received by children in this group (17.8), the renal Hg concentration associated with incipient increases in urinary porphyrins was estimated to be approximately 2.7 microg/g renal cortex. This value corresponds to an observed mean urinary Hg concentration of 3.2 microg/g creatinine, which is approximately fivefold less than that at which renal damage from Hg exposure is estimated to occur in children. These findings are consistent with growing evidence supporting the sensitivity of urinary porphyrins as a biological indicator of subclinical Hg exposure in children. PMID- 19557619 TI - Evaluation of potassium iodide (KI) and ammonium perchlorate (NH4ClO4) to ameliorate 131I- exposure in the rat. AB - Nuclear reactor accidents and the threat of nuclear terrorism have heightened the concern for adverse health risks associated with radiation poisoning. Potassium iodide (KI) is the only pharmaceutical intervention that is currently approved by the Food and Drug Administration for treating (131)I(-) exposure, a common radioactive fission product. Though effective, KI administration needs to occur prior to or as soon as possible (within a few hours) after radioactive exposure to maximize the radioprotective benefits of KI. During the Chernobyl nuclear reactor accident, KI was not administered soon enough after radiation poisoning occurred to thousands of people. The delay in administration of KI resulted in an increased incidence of childhood thyroid cancer. Perchlorate (ClO(4)(-)) was suggested as another pharmaceutical radioprotectant for 131I- poisoning because of its ability to block thyroidal uptake of iodide and discharge free iodide from the thyroid gland. The objective of this study was to compare the ability of KI and ammonium perchlorate to reduce thyroid gland exposure to radioactive iodide (131I-). Rats were dosed with 131I- tracer and 0.5 and 3 h later dosed orally with 30 mg/kg of either ammonium perchlorate or KI. Compared to controls, both anion treatments reduced thyroid gland exposure to 131I- equally, with a reduction ranging from 65 to 77%. Ammonium perchlorate was more effective than stable iodide for whole-body radioprotectant effectiveness. KI-treated animals excreted only 30% of the (131)I(-) in urine after 15 h, compared to 47% in ammonium perchlorate-treated rats. Taken together, data suggest that KI and ammonium perchlorate are both able to reduce thyroid gland exposure to 131I- up to 3 h after exposure to 131I-. Ammonium perchlorate may offer an advantage over KI because of its ability to clear 131I- from the body. PMID- 19557620 TI - Demographic, residential, and behavioral determinants of elevated exposures to benzene, toluene, ethylbenzene, and xylenes among the U.S. population: results from 1999-2000 NHANES. AB - Volatile organic compounds (VOC) represent a broad spectrum of compounds and there is growing concern that VOC exposures, in addition to increasing risks for cancer, may be implicated in exacerbating asthma and other adverse respiratory effects. Yet little is known about exposures in the U.S. population beyond the seminal Total Exposure Assessment Methodology (TEAM) studies that were conducted by the U.S. Environmental Protection Agency (U.S. EPA) between 1979 and 1987. This investigation was carried out to evaluate the relationship between personal exposures to benzene, toluene, ethylbenzene, and xylenes (BTEX) and socioeconomic, behavioral, demographic, and residential characteristics using a subsample from the National Health and Nutrition Examination Survey (NHANES) (636 participants who represented an estimated 141,363,503 persons aged 20 to 59 yr in the United States). Personal VOC exposures were evaluated using organic vapor monitors for periods that ranged from 48 to 72 h, and participants were administered a questionnaire regarding personal behaviors and residential characteristics while wearing the monitor. Geometric mean (GM) levels were significantly higher for males for all compounds except toluene. For benzene, GM levels were elevated among smokers and Hispanics. Sociodemographic characteristics could not be evaluated simultaneously in the weighted multiple regression models with the VOC questionnaire data because of issues associated with multicollinearity. Results from the regression analyses suggest that the presence of an attached garage (BTEX), having windows closed in the home during the monitoring period (benzene, toluene), pumping gasoline (toluene, ethylbenzene, and xylenes), or using paint thinner, brush cleaner, or stripper (xylenes) results in higher exposure in the general population and confirm previous findings of studies that were more regional in scope. Once the complete NHANES VOC data are released, additional study is warranted to explore whether risk factors associated with elevated VOC exposures differ in subgroups of U.S. adults, which should inform efforts to develop approaches for minimizing VOC exposures and ameliorating environmental health risks. PMID- 19557621 TI - Perceptual asymmetries influence task choice: the effect of lateralised presentation of hierarchical stimuli. AB - The current study examined how hemispheric asymmetries in perceptual processing affect control processes associated with voluntary task choice during multitask behaviour. In a voluntary task-switching paradigm, where participants are free to choose which task to perform on each trial, participants identified either the global-or local-level features of hierarchical stimuli presented to either the left or right visual field. Hemispheric asymmetries in perception of lateralised hierarchical stimuli were evident in reaction times. Importantly, participants were more likely to categorize the stimulus in line with the processing efficiency of the hemisphere to which it was initially presented: the global task for left visual field presentation and the local task for right visual field presentation. Perceptual processing characteristics influence control processes associated with task choice in multitask environments. PMID- 19557622 TI - Influence of MBL-2 mutations in the infection risk of patients with follicular lymphoma treated with rituximab, fludarabine, and cyclophosphamide. AB - The employment of current treatments based on chemotherapy and immunotherapy leads to inmunosuppression. The presence of mutations or polymorphisms in genes related to immune system might involve an additional disadvantage. The aim of the present study was to analyze mannose-binding lectin (MBL-2 gene) mutations and their association with severe infections and event-free survival in patients diagnosed with follicular lymphoma, treated uniformly, in the clinical trial LNHF 03. The results of this trial showed impressive clinical efficacy but was complicated with 80 documented infectious episodes. Patients were classified into two genotypic groups, AA and AO/OO, based on their haplotypic inheritance. Neither the number of infectious episodes nor differences in event-free survival was found as a function of MBL-2 groups. Other factors, including the lymphoma malignancy and the immune alterations associated with the disease, should be considered responsible for this observation. PMID- 19557624 TI - t(14;18) cells are getting hard to find. PMID- 19557623 TI - Complete remissions observed in acute myeloid leukemia following prolonged exposure to lintuzumab: a phase 1 trial. AB - A multi-institutional, phase 1 dose-escalation trial of lintuzumab (humanized anti-CD33 antibody; SGN-33, HuM195) was performed in patients with CD33-positive myeloid malignancies. In this study, higher doses than previously tested and prolonged duration of treatment for responding patients were evaluated. Over the dose range of 1.5-8 mg/kg/week, lintuzumab was well tolerated, and a maximum tolerated dose was not defined. The most common adverse event was transient chills with the initial lintuzumab infusion (39%). Responses were observed in 7 of 17 patients with acute myeloid leukemia: morphologic complete remission (n = 4), partial remission (n = 2), and morphologic leukemia-free state (n = 1). Of 14 patients with myelodysplastic syndrome or myeloproliferative diseases, 1 patient had major hematologic improvement and 9 patients had stable disease. In contrast to aggressive conventional chemotherapy, lintuzumab was administered in an ambulatory clinic setting with acceptable toxicity. PMID- 19557625 TI - 'Rapping' on the door of drug resistance. PMID- 19557626 TI - The stromal derived factor-1?CXCR4 axis--a legitimate therapeutic target in multiple myeloma? PMID- 19557627 TI - BH3-mimetics--the solution to chemoresistance? PMID- 19557628 TI - Therapy-related acute promyelocytic leukemia: further insights into the molecular basis of the disease and showing the way forward in therapy. PMID- 19557629 TI - B cell activator factor and a proliferation-inducing ligand at the cross-road of chronic lymphocytic leukemia and autoimmunity. AB - The combination of the neoplastic accumulation of mature B lymphocytes with the presence of autoimmune phenomena is a characteristic finding in chronic B cell lymphoproliferative disorders, particularly chronic lymphocytic leukemia. Identification of mechanisms linking neoplasia to the autoimmune defects is important for a better understanding and improving the treatment of these conditions. Among such mechanisms, the B cell activator factor (BAFF) and a proliferation-inducing ligand (APRIL), two members of the tumor necrosis factor family, play an important role. BAFF and APRIL have both been associated with autoimmunity, with their underlying mechanism of action most likely being related to the rescue of autoreactive B cells. In addition, BAFF and APRIL are crucial in B cell development and homeostasis particularly via the activation of NF-kappaB pathway-mediated survival signals. These two proteins, therefore, constitute a paradigm of pathophysiological defects linking neoplasia and autoimmunity, thereby providing a better understanding of chronic B cell lymphoproliferative disorders. PMID- 19557630 TI - The persistence of t(14;18)-bearing cells in lymph nodes of patients with follicular lymphoma in complete remission: the evidence for 'a lymphoma stem cell'. AB - Monitoring of t(14;18) in blood or bone marrow in follicular lymphoma (FL) remains controversial. We attempted to monitor t(14;18) in lymph nodes by ultrasound-guided fine needle aspirations (UG-FNA). First, we confirmed t(14;18) in 27/31 UG-FNAs of lymph nodes with fluorescent in situ hybridisation (FISH) and/or polymerase chain reaction (PCR) in patients with advanced disease. In complete (CR) and molecular remission, there were repeated 18 UG-FNAs in 17 patients. Five of 18 UG-FNA were technically unsuccessful and 6/18 samples contained fibrosis. Despite that, these patients had a better prognosis. In 7/7 aspirations in six patients, t(14;18) was detected. Three patients are still in CR, even one of them remains in long lasting remission despite two consecutive evidences of t(14;18) in UG-FNA. Another three of these patients relapsed a few months after UG-FNA. This study is proof of the principle of the detection of residual t(14;18) bearing cells in previously involved lymph nodes despite patients being in remission. PMID- 19557631 TI - A retrospective study to compare two methotrexate-based regimens for primary central nervous system lymphoma. AB - A retrospective analysis to compare the treatment outcomes of two regimens with different doses of methotrexate (MTX). Seventy-two patients, newly diagnosed with primary central nervous system lymphoma between 1995 and 2006, were included. All patients were treated with one of the two different MTX regimens depending on when the diagnosis was made. Thirty-six patients diagnosed between 1995 and 2002 were treated with 1 g/m(2) of intravenous MTX (HD-MTX 1 g/m(2), cohort 1). The other 36 patients, diagnosed between 2003 and 2006, received 3.5 g/m(2) of intravenous MTX (HD-MTX 3.5 g/m(2), cohort 2). The median age was 47 years (range, 17-78 years) and 42 patients (58.3%) were male. The median overall survival (OS) and progression-free survival (PFS) of all patients was 90.3 and 52.9 months, respectively. Although OS and PFS was not statistically different between the two cohorts, cohort 2 achieved higher complete response/unconfirmed complete response rates than cohort 1 at an evaluation conducted between completion of intravenous MTX therapy and the initiation of radiotherapy (52.8% vs. 16.7%, respectively; p = 0.005). Furthermore, there were no deaths within 6 months of MTX therapy for the cohort 2, whereas there were eight deaths by 6 months for cohort 1 (p = 0.003). Even though cohort 2 failed to show superior survival outcomes compared with cohort 1 after sequential brain radiotherapy and intravenous cytarabine, the higher early CR/CRu rate of cohort 2 compared with cohort 1 might indicate that a high dose of MTX is desirable. PMID- 19557632 TI - Clinical features and treatment outcomes of adult B- and T-lymphoblastic lymphoma: results of multicentre analysis in Korea. AB - We performed a retrospective multicentre analysis to study the clinical features and treatment outcomes of B-lymphoblastic lymphoma (B-LBL) and T-lymphoblastic lymphoma (T-LBL) in Asian adult patients, and identify risk factors that predict relapse and poor prognosis. Fifty-five newly diagnosed patients (45 T-LBL and 10 B-LBL) were analysed. All patients were treated with intensive chemotherapy regimens including VPDL (vincristine, prednisolone, daunorubicin, L asparaginase), CALGB (Cancer and leukemia group B), and Stanford/Northern California Oncology Group (NCOG). There was no difference of clinical features between B- and T-LBL except the frequent site of involvement. The overall response rate including complete response (28/55, 50.9%) and partial response (18/55, 32.7%) was 83.6%. Among 46 responders, 22 patients relapsed leading to 20 deaths. Partial responders showed more frequent relapse (10/18, 55.6%) than complete responders (11/28, 39.2%). The median progression-free survival (PFS) was 17 months (95% confidence interval, 11.5-22.5), and the 2-year overall survival was 52 +/- 7% with a median follow-up of 50 months (range 8-152). Treatment outcome of T-LBL and B-LBL was not significantly different in terms of response and survival. The presence of pleural effusion was significantly prognostic for overall and PFS (p < 0.05). In conclusion, clinical features and treatment outcome of Asian adult LBL were comparable to previous results, and the prognosis is still poor despite intensive chemotherapy. PMID- 19557633 TI - A prognostic scoring system for adult patients less than 60 years of age with acute lymphoblastic leukemia in first relapse. AB - The outcome of patients with acute lymphoblastic leukemia (ALL) in first relapse is poor. We retrospectively evaluated patients with ALL in first relapse, 18-60 years of age, to define a prognostic score. For all patients, a scoring system of 0-3 was developed with 1 point for each of the following: age at diagnosis >or=45 years, lactate dehydrogenase (LDH) at the time of relapse >or=1.5 times upper limits of normal (ULN), not proceeding to allogeneic bone marrow transplant (BMT). A similar scoring system was developed for patients proceeding to BMT. LDH >or=1.5 times ULN at the time of relapse predicted poor overall survival. Patients with a prognostic score of greater than 1 have a poor prognosis, even with BMT, and should be considered for treatment with innovative approaches such as Phase 1 clinical trials. PMID- 19557634 TI - Improved treatment results in Mexican children with acute myeloid leukemia using a Medical Research Council (MRC)-acute myeloid leukemia 10 modified protocol. AB - We analysed the results of three protocols from 1990 to 2005. Protocol I (1990 1996) consisted of a 2 year VAPA regime. Protocol II (1996-2003) on 1 year daunorubicin/cytarabine alternating with etoposide/cytarabine. Protocol III (2003 2005) on six cycles MRC AML 10 modified. Patients with de novo acute myeloid leukemia 0 to 18 years were included. Demographic and clinical characteristics were analysed. Patients with >100,000 leukocytes, M4 or M5 and primary CNS disease were considered high risk. We compared remission rate, overall and event free survival. Descriptive statistics, chi square, Kaplan-Meier and long rank tests were used. One hundred forty-five patients were included, 46 in Protocol I; 60 in II and 39 in III. There were no differences in characteristics between groups, except for more low risk patients in Protocol II (61%vs. 43% and 41%. (p = 0.05). Remission rate for Protocol I was 52%, for II 50% and for III 92% (p = 0.0001). Relapse was 18, 30 and 35, respectively (p = 0.141). Five-year event free survival was 17.9% +/- 6.6%, 15.5% +/- 4.1% and 43.5% +/- 4.1% (s.e) (p = 0.0002). Five-year overall survival was 19.5% +/- 8%, 17.2% +/- 5.9% and 51.2% +/ 4.1% (s.e) (p = 0.0002). The results were superior in the MRC-10 derived protocol. PMID- 19557635 TI - Acute leukemia among the adult population of United Arab Emirates: an epidemiological study. AB - There is no published data regarding adult acute leukemia (AL) in the United Arab Emirates (UAE). Our objectives were to determine the distribution and incidence of adult AL in UAE (nationals and non-nationals). This epidemiological survey recovered 263 adult patients with AL diagnosed between January 2000 and December 2006 with a median age of 34 years. Twenty-four percent were UAE nationals and 63% were males. Acute lymphoblastic leukemia (ALL) was more frequently diagnosed (32%) than in western countries. This clearly reflects the population structure of the UAE which consists of predominantly young males. There is a tendency for lower crude and age-specific incidence rates of AL, acute myeloid leukemia (AML) and ALL in the UAE when compared with those in western countries. We found a statistically significant higher incidence of AML among national females than in national males (p = 0.04). This is reflected in a significantly higher incidence of AL (p = 0.02) and AML (p = 0.02) among the females when compared with the males in the total population of the UAE. This result contradicts the generally known finding that AML and ALL are more common in males. The implication of cumulative risk factors to which females could be exposed, such as vitamin D deficiency as a result of sunlight deprivation and direct exposure to benzene and color enhancement chemicals in henna, could not be excluded and warrant further investigation. PMID- 19557636 TI - New mutations detected by denaturing high performance liquid chromatography during screening of exon 6 bcr-abl mutations in patients with chronic myeloid leukemia treated with tyrosine kinase inhibitors. AB - Point mutations within the ABL kinase domain are the most frequent mechanism for reactivation of kinase activity of the BCR-ABL gene and have been associated with clinical resistance to tyrosine kinase (TK) inhibitors in patients with CML, conferring a poor prognosis. T315I (Treonine-->Isoleucine) is a mutation in the exon 6 of BCR-ABL gene that makes the protein resistant to kinase inhibitors currently used for treating CML. Denaturing High-performance liquid chromatography (D-HPLC) allows for high throughput mutation screening. In this study, we screened mutations in exon 6 of the BCR-ABL gene in patients presenting failure or sub optimal response according to Leukemia Net criteria and correlated the presence of mutations with clinical outcome. Genomic DNA was extracted from peripheral blood samples from 93 patients with CML (5 intolerant and 88 resistant). The PCR product was analysed by D-HPLC, and the patients samples with abnormal D-HLPC profiles were submitted to automated sequencing, using specific primers. Overall survival (OS) was calculated from the date of mutation analysis, for the whole group and for both groups (mutation versus no mutation). We screened mutations in exon 6 of the BCR-ABL gene in 93 CML TKI - resistant patients. Twenty-three out of 93 samples (25%) showed an abnormal elution profile. Automated sequencing confirmed the presence of a nucleotide change in 19 out of 23 cases: one polymorphism, T315T, seven known point mutations: T315I, F317L, V339L, M351T, E355G and F359V and three novel mutations: C305R, D325D and I360S. OS for the whole group was 80% in a median observation time of 30 months. OS for patients without the mutation was 87% and with the mutation was 56%, in a median observation time of 37 and 10 months, respectively (p < 0.0001, RR = 68). D-HPLC is a practical and sensitive method for routine clinical monitoring for emergence of kinase domain mutations and may be useful for optimising therapy in CML. The screening of mutations in exon 6 is clinically relevant, once the presence of mutations confers a poor outcome. Early detection of emerging mutant clones may help in decision-making for alternative treatment. PMID- 19557637 TI - Targeting mammalian target of rapamycin to both downregulate and disable the P glycoprotein pump in multidrug-resistant B-cell lymphoma cell lines. AB - Previous studies have shown that rapamycin can inhibit the growth of several different types of human tumor cells in vitro. In certain cases, it can reverse the phenotype of multidrug resistant (MDR) cells. However, there is limited information concerning its effect on P-glycoprotein (P-gp), a pump that is responsible for chemoresistance in many MDR cells. We investigated the effect of rapamycin on both P-gp function and the MDR phenotype in four cell lines. One cell line was also xenografted into SCID mice to determine whether rapamycin would chemosensitize the cells in vivo. Because rapamycin targets the mammalian target of rapamycin (mTOR) pathway, we also used our cells to confirm that rapamycin modified the expression of mTOR and effectively suppressed the phosphorylation of two downstream effector molecules in the mTOR pathway, S6K1, and 4E-BP1. We demonstrated that it inhibited the growth of the three cell lines in vitro and one in vivo showing that it modulated both the expression and function of P-gp and chemosensitized the three cell lines as effectively as verapamil. PMID- 19557638 TI - Targeting the Bcl-2 family of proteins in Hodgkin lymphoma: in vitro cytotoxicity, target modulation and drug combination studies of the Bcl-2 homology 3 mimetic ABT-737. AB - With currently available treatment, patients with refractory Hodgkin lymphoma (HL) or those who relapse multiple times have an extremely poor prognosis. Therefore, new agents and novel therapeutic approaches are urgently needed. Anti apoptotic proteins such as Bcl-2 and Bcl-x have been associated with the growth and survival of Hodgkin Reed-Sternberg cells and are potential therapeutic targets. ABT-737 is a small molecule that inhibits the Bcl-2 family of apoptosis regulators. In this study, we show the concentration-dependent and time-dependent cytotoxicity of ABT-737 against cell lines derived from patients with HL. A concurrent reduction in a number of intracellular cell growth and survival related molecules, such as Bcl-2, Bcl-xl, NF-kappaB and survivin was also seen. Drug combination studies using a panel of conventional and novel therapeutic agents show that ABT-737 potentiates the activity of agents that have inherent anti-lymphoma activity and provide support for the evaluation of ABT-737 in the clinical setting. PMID- 19557640 TI - Preferential expression of phosphatidylglucoside along neutrophil differentiation pathway. AB - Phosphatidylglucoside (PtdGlc), a new type of glycolipid, was recently identified. We examined PtdGlc expression in normal blood cells and leukemic cells using an anti-PtdGlc monoclonal antibody, DIM-21. Neutrophils, monocytes, HL-60 cells and a subset of cord blood (CB) CD34(+) cells, but not erythroblasts, expressed lipid antigen. PtdGlc was preferentially expressed along the neutrophil differentiation pathway of CB CD34(+) cells treated with cytokines and HL-60 cells treated with retinoic acid. PtdGlc expression was not increased in HL-60 cells treated with phorbol ester. CB CD34(+) cells contained a population of PtdGlc(+) cells, and CB CD34(+)PtdGlc(+) cells produced mainly granulocyte macrophage colonies and a small number of erythroid colonies. A positive correlation between PtdGlc expression and CD15 expression in leukemic cells from patients with acute myeloblastic leukemia was shown. These results indicate that increasing PtdGlc expression is seen with neutrophil maturation. PMID- 19557641 TI - Lymphoblastic T-cell lymphoma in mice is unaffected by Celecoxib as single agent or in combination with cyclophosphamide. AB - Celecoxib, an inhibitor of cyclooxygenase-2, is a promising novel antitumor agent with pleitropic mechanisms of action. Whereas this drug induces growth arrest and apoptosis of B-lymphoma cells, its effect against aggressive T-cell neoplasms remains to be studied. We therefore evaluated Celecoxib therapy of immunocompetent mice transplanted with lymphoblastic T-cell lymphomas. Oral Celecoxib in clinically relevant and non-toxic doses did not affect the degree of hypersplenism or the number of viable lymphoma cells. The clinical deterioration of Celecoxib-treated mice was not different from untreated controls. The impact of adding Celecoxib (60 mg/kg) to cyclophosphamide (200 mg/kg x 1, i.p.) was assessed but showed no benefit compared to cyclophosphamide alone. Thus, Celecoxib lacks effect against lymphoblastic T-cell lymphoma in mice. PMID- 19557639 TI - Function of retinoid acid receptor alpha and p21 in all-trans-retinoic acid induced acute T-lymphoblastic leukemia apoptosis. AB - All-trans-retinoic acid (ATRA) is a morphogenetic signalling molecule derived from vitamin A and is used clinically to target acute promyelocytic leukemia by inducing differentiation of immature blood cells. Retinoid signals are mediated by retinoic acid (RA) receptors (RARs) and retinoid X receptors (RXRs). Retinoic acid receptors consist of RARalpha, RARbeta and RARgamma isotypes. Among these components, RARalpha is preferentially bound to ATRA, which is used to treat acute T-lymphoblastic leukemia, yet the conditions and mechanisms remain unknown. In this study, we have demonstrated that, in human acute T-lymphoblastic leukemia Molt3 cells, inhibition of RA-induced proliferation results from massive cell death characterised by apoptosis. The effect of ATRA:RARalpha binding on apoptosis in Molt3 cells has been investigated. Consequently, it has been shown that, in RA-treated Molt3 cells, upregulation of p21 due to RA accompanies caspase 3/PARP activation which precedes the occurrence of apoptosis. PMID- 19557642 TI - Proteasome inhibitor MG-132 mediated expression of p27Kip1 via S-phase kinase protein 2 degradation induces cell cycle coupled apoptosis in primary effusion lymphoma cells. AB - Primary effusion lymphoma (PEL) is an incurable, aggressive B-cell malignancy that develops rapid resistance to conventional chemotherapy. MG-132, a proteasome inhibitor, suppresses cell proliferation and induces apoptosis in several PEL cell lines. Treatment of PEL cells with MG-132 results in downregulation of S phase kinase protein 2 (SKP2) and accumulation of p27Kip1. Furthermore, MG-132 treatment of PEL cells causes Bax conformational changes, leading to loss of mitochondrial membrane potential and release of cytochrome c to the cytosole. Such cytochrome c release results in sequential activation of caspases and apoptosis, while pretreatment of PEL cells with universal inhibitor of caspases, z-VAD-fmk prevents cell death induced by MG-132. Finally, our data demonstrated in PEL cells that MG-132 downregulates the expression of inhibitor of apoptosis proteins XIAP, cIAP1 and survivin. Altogether, these findings suggest that MG-132 is a potent inducer of apoptosis of PEL cells via downregulation of SKP2 leading to accumulation of p27Kip1, resulting in cell cycle arrest and apoptosis and strongly suggest that targeting the proteasomal pathway may provide a novel therapeutic approach for the treatment of PEL. PMID- 19557643 TI - Acute promyelocytic leukemia after Stanford V plus radiotherapy for advanced Hodgkin lymphoma. PMID- 19557644 TI - Primary intestinal diffuse large B-cell lymphoma presenting as multiple lymphomatous polyposis. PMID- 19557645 TI - S-phase fraction as response marker in patients with chronic myeloid leukemia. PMID- 19557646 TI - Syndrome of inappropriate antidiuretic hormone associated with chemotherapy induced tumor lysis in a patient with peripheral T-cell lymphoma unspecified. PMID- 19557647 TI - Clofarabine in relapsed lymphoma: what is the optimal dose? PMID- 19557650 TI - Isolation and identification of Rhizomucor pusillus from pleural zygomycosis in an immunocompetent patient. AB - Zygomycosis is usually an invasive mycotic disease caused by fungi in the class Zygomycetes. It often occurs in immunocompromised patients, but sporadic cases without apparent immune impairment have been described. This report presents the first case of pleural zygomycosis caused by Rhizomucor pusillus, an uncommon pathogen of human infection. A 19-year-old man was found to have pleuritis several days after a drainage catheter was implanted to cure a pneumothorax caused by a ruptured bulla. Local pneumonectomy to resect the ruptured bulla and vacuuming of the pleural fluid was performed. Rhizomucor pusillus was cultured from the pleural fluid and irregular broad sparsely septate hyphae, consistent with zygomycetes, were histologically detected in the thickened pleura of the resected bulla. The catheter was suspected of having been contaminated with the fungus, but no evidence could be obtained. His fungal pleuritis subsided without any antifungal medical therapy and his immunocompetence seemed to contribute to limiting the infection. PMID- 19557652 TI - Management of corm-rot disease of Gladiolus by plant extracts. AB - A pot experiment was conducted to evaluate the efficacy of aqueous extracts of six plant species, namely Azadirachta indica A. Juss. (neem), Alstonia scholaris (L.) R. Br., Lawsonia alba Lam., Allium cepa L., A. sativum L. and Zingiber officinale Roscoe, and a systemic fungicide carbendazim 50% (w/w) WP, to manage the corm-rot disease of Gladiolus (Gladiolus grandiflorus L.) caused by a fungal pathogen Fusarium oxysporum f.sp. gladioli (Massey) Snyd. & Hans. Fusarium inoculation showed 80% disease incidence with 54 disease lesions per corm. Recommended dose of the chemical fungicide carbendazim significantly reduced the disease incidence to 13% and number of lesions to six per corm. Plant extract treatments exhibited variable effects on the incidence and severity of the disease. In general, all the test plant extracts managed the corm-rot disease to some extent. Aqueous bulb extracts of A. sativum and A. cepa and the rhizome extract of Z. officinale showed better disease management potential than that of the recommended dose of carbendazim. Fusarium inoculation significantly declined shoot growth. In general, carbendazim, as well as aqueous extracts, enhanced shoot growth to variable extents as compared to the Fusarium control. PMID- 19557653 TI - Clinical manifestation and molecular genetic characterization of MYH9 disorders. AB - Currently, the May-Hegglin anomaly (MHA), Sebastian (SBS), Fechtner (FTNS) and Epstein (EPS) syndrome are considered to be distinct clinical manifestations of a single disease caused by mutations of the MYH9 gene encoding the heavy chain of non-muscle myosin IIA (NMMHC-IIA). Manifestations of these disorders include giant platelets, thrombocytopenia and combinations of the presence of granulocyte inclusions, deafness, cataracts and renal failure. We examined 15 patients from 10 unrelated families on whom we performed immunostaining of NMMHC-IIA in blood samples. Polymerase chain reaction (PCR) analysis of selected exons of the MYH9 gene revealed mutations in nine samples with one novel mutation. Results of fluorescence and mutational analysis were compared with clinical manifestations of the MYH9 disorder. We also determined the number of glycoprotein sites on the surface of platelets. Most patients had an increased number of glycoproteins, which could be due to platelet size. PMID- 19557654 TI - Correspondence regarding the use of the acronym HAM. PMID- 19557655 TI - Age influences anthropometric and fitness-related predictors of bone mineral in men. AB - OBJECTIVE: This study assessed the influence of age on the predictors of bone mineral in men. METHODS: Middle-age (n = 41, 54 +/- 4 yrs) and older (n = 40, 69 +/- 5 yrs) men underwent grip and knee extensor strength tests, total body dual energy X-ray absorptiometry with regional analyses and a graded exercise treadmill test. RESULTS: Bone-free lean mass (BFLM) and, to a lesser extent, fat mass (FM) were correlated with bone mineral variables in middle-age men. In older men, BFLM and, to a lesser extent, FM were related to bone mineral content (BMC) at most sites, but inconsistently to bone mineral density (BMD). Knee extensor strength related to bone mineral (BMC and BMD) at most sites in middle-age men, but none in older men. Grip strength inconsistently related to bone mineral in both groups. Aerobic capacity related to bone mineral in middle-age men, but none in older men. In multiple regression, body weight or BFLM predicted bone mineral in middle-age men (R2 = 0.33-0.68) and BMC in older men (R2 = 0.33-0.50). Predictors of BMD were inconsistent in older men. CONCLUSIONS: Relationships of body composition, muscular strength and aerobic capacity to bone mineral are stronger in middle-age versus older men. PMID- 19557656 TI - Lesion location and repeatable battery for the assessment of neuropsychological status performance in acute ischemic stroke. AB - The validity of the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) was evaluated in a sample of acute ischemic stroke patients. A total of 164 ischemic stroke patients with anterior fossa lesions were divided into groups according to lesion laterality (left, right, or bilateral) and location (cortical versus subcortical) as determined by CT and/or MRI findings. The hypotheses for this study were: (1) that left hemispheric stroke patients would perform better than their counterparts on the Visuospatial/Constructional and Attention Indexes; (2) that right hemisphere stroke patients would outperform their counterparts on the Immediate Memory, Delayed Memory, and Language Indexes; (3) that patients with subcortical lesions would outperform those with cortical lesions on the Language, Immediate Memory, and Delayed Memory indexes; and (4) that patients with cortical lesions would outperform those with subcortical lesions on the Attention and Visuospatial/Constructional Indexes. A multivariate analysis of variance (MANOVA) disclosed significant main effects for both lesion side and location, with no location by side interaction. Group comparisons of the five RBANS index scores disclosed modest effects for side of lesion, with right hemisphere patients outperforming those with left sided and bilateral lesions on the Immediate and Delayed Memory, and Language Index Scores. Right hemisphere stroke patients also outperformed left and bilateral lesion patients on the Attention Index. The left hemisphere patients outperformed the right hemisphere and bilateral patients on the Visuospatial/Constructional Index. The effect for location was significant only for the Visuospatial/Constructional Index where the subcortical patients outperformed the cortical patients. PMID- 19557659 TI - The significance of fetal renal pelvic dilatation as a predictor of postnatal outcome. AB - OBJECTIVES: To define the clinical outcome of fetal renal pelvic dilatation (FRPD) in cohort of infants in United Arab Emirates. STUDY DESIGN: Data were collected from all fetuses having FRPD from January 2005 to February 2008. FRPD was graded as normal (<5 mm), mild (5-9 mm), moderate (10-15 mm), and severe (>15 mm). RESULTS: Data from 80 fetuses with 120 kidneys were studied. Nine resolved antenatally and seven were lost to follow up. Of the remaining 89 FPRD (64 patients), 36% had normal postnatal ultrasound, 22.5% significant uropathy, and 41.5% had isolated hydronephrosis. Pelvi-ureteric junction obstruction was the commonest identified underlying abnormality. Severe FRPD predicted significant postnatal uropathy with a sensitivity of 65% and a specificity of 98.6%. Moderate FRPD increased the sensitivity to 95% but decreased the specificity to 60.9%, mild FPRD was seldom (4%) associated with significant postnatal pathology. Postnatal resolution was significantly (p = 0.01) higher in mild RPD than in the moderate or severe group. CONCLUSION: Severe FRPD need comprehensive postnatal assessment. Although moderate FRPD had a high prevalence of uropathy, they rarely needed surgical intervention. Parents could be reassured that RPD of less than 10 mm in the third trimester is unlikely to be associated with significant uropathology. PMID- 19557660 TI - Undiagnosed maternal phenylketonuria: own clinical experience and literature review. AB - Phenylketonuria (PKU) is an autosomal recessive inborn error of phenylalanine (Phe) metabolism resulting from deficiency of phenylalanine hydroxylase (PAH). Most forms of PKU are caused by mutations in the PAH gene. Untreated PKU is associated with an abnormal phenotype, which includes growth failure, seizures, global developmental delay and severe intellectual impairment. The maternal PKU (MPKU) syndrome is caused by high blood Phe concentrations during pregnancy and presents with serious foetal anomalies, especially microcephaly, congenital heart disease and mental retardation. However, since the introduction of newborn screening programs and with early dietary intervention, children born with PKU can now expect to lead relatively normal lives. We present the case of a 33-year old woman who had been diagnosed as having PKU only after a pregnancy with MPKU embryopathy, to emphasize that undiagnosed maternal phenylketonuria still exists. On that ground, we reviewed updated literature on the pathogenesis of this syndrome, possibility of prophylaxis and treatment. PMID- 19557662 TI - Maternal hemoglobin concentration and its association with birth weight in newborns of mothers with preeclampsia. AB - OBJECTIVE: Maternal hemoglobin concentration is inversely related to newborn size presumably through plasma volume constriction. We sought to determine whether birth weight would show an inverse relationship to hemoglobin concentration in a group of infants whose mothers had preeclampsia, where plasma volume constriction is common. METHODS: Electronic and paper chart review identified 142 nulliparous women with preeclampsia (excluding hemolysis, elevated liver enzymes, low platelets syndrome). Birth weight percentile was determined based on cross sectional hybrid growth curves. Maximal third trimester maternal hemoglobin concentrations were obtained and standardised to z-scores based on gestational age matched normative data. Birth weight percentile was examined as a function of hemoglobin z-score using appropriate statistics. RESULTS: Average gestational age at delivery was 35.9 +/- 1.9 weeks. Mean birth weight percentile for infants of preeclamptic mothers was 34 +/- 32. Mean hemoglobin z-score for mothers with preeclampsia was 0.3 +/- 1.5, significantly higher than a control population (p = 0.04). Maternal hemoglobin z-score was inversely associated with birth weight percentile (r = -0.18, p = 0.03). CONCLUSION: Maternal hemoglobin concentrations are significantly elevated prior to delivery in women with preeclampsia. There is a statistically significant inverse correlation of maternal hemoglobin concentration to birth weight percentile. PMID- 19557664 TI - Neonatal outcomes in the setting of preterm premature rupture of membranes complicated by chorioamnionitis. AB - OBJECTIVE: To examine the outcomes of neonates born to women with chorioamnionitis in the setting of preterm premature rupture of membranes (PPROM). METHODS: A retrospective cohort study was conducted of deliveries with diagnosis of PPROM between 24 and 34 weeks of gestation at an academic medical center. Patients who delivered with the diagnosis of clinical chorioamnionitis were compared with patients who delivered without this diagnosis. Neonatal outcomes including Apgar scores, intracranial hemorrhage (ICH), sepsis, pneumonia, respiratory distress syndrome (RDS), and necrotizing enterocolitis (NEC) were assessed. Dichotomous outcomes were compared using chi-square test. Multivariable regression analyses were performed to control for potential confounding variables. RESULTS: Of the 1153 patients diagnosed with PPROM, 29.0% were diagnosed with chorioamnionitis prior to delivery. Neonates born to mothers with a diagnosis of chorioamnionitis in the setting of PPROM had higher incidences (34.8%) of low 5-min Apgar scores, RDS, NEC, ICH, and pneumonia compared with 22.9% in neonates born to mothers without chorioamnionitis (p < 0.001). CONCLUSIONS: Patients who develop chorioamnionitis in the setting of PPROM are at higher risk for adverse neonatal outcomes compared with patients without chorioamnionitis in the setting of PPROM. PMID- 19557663 TI - Recurrence of fetal growth restriction in singleton and twin gestations. AB - OBJECTIVE: Patterns of recurrence of restricted fetal growth provide important insights to understand the relative contributions of genetic versus environmental influences. Although there is evidence of increased tendency of small for gestational age (SGA) births to recur, whether similar patterns of recurrence in twins among women that delivered a prior singleton SGA birth remains poorly studied. METHODS: We used Missouri's maternally-linked data (1978-1997), and restricted the analysis to women that delivered their first two consecutive live births. SGA (birthweight <10th and <5th centile for gestational age) recurrence was examined in two distinct analyses: women that delivered their first two singleton live births (n = 305,654) and those that delivered their first singleton live birth followed by twin live births (n = 8594). Sib-sib pairwise odds ratio (pOR) were estimated from bivariate logistic regression with robust variance estimation after adjustments for confounders. RESULTS: Risks of SGA were 24.3% and 6.1% in the second singleton birth among women with and without a previous singleton SGA, respectively (pOR 3.9, 95% CI 3.7-4.0). The corresponding risks among twins with and without a previous singleton SGA birth were 16.9% and 6.7%, respectively (pOR 2.3, 95% CI 1.8-2.8). In the singleton-singleton cohort, the highest recurrence risk for SGA occurred around the same gestational age window as the first singleton SGA birth. These associations were stronger for more severe forms of SGA (<5th centile). CONCLUSIONS: The likelihood of SGA to recur within sibships is strong, with varying magnitude of risks between singleton-singleton and singleton-twin births. PMID- 19557666 TI - Seeing the future: Natural image sequences produce "anticipatory" neuronal activity and bias perceptual report. AB - This paper relates human perception to the functioning of cells in the temporal cortex that are engaged in high-level pattern processing. We review historical developments concerning (a) the functional organization of cells processing faces and (b) the selectivity for faces in cell responses. We then focus on (c) the comparison of perception and cell responses to images of faces presented in sequences of unrelated images. Specifically the paper concerns the cell function and perception in circumstances where meaningful patterns occur momentarily in the context of a naturally or unnaturally changing visual environment. Experience of visual sequences allows anticipation, yet one sensory stimulus also "masks" perception and neural processing of subsequent stimuli. To understand this paradox we compared cell responses in monkey temporal cortex to body images presented individually, in pairs and in action sequences. Responses to one image suppressed responses to similar images for approximately 500 ms. This suppression led to responses peaking 100 ms earlier to image sequences than to isolated images (e.g., during head rotation, face-selective activity peaks before the face confronts the observer). Thus forward masking has unrecognized benefits for perception because it can transform neuronal activity to make it predictive during natural change. PMID- 19557667 TI - The role of working memory in auditory selective attention. AB - A growing body of research now demonstrates that working memory plays an important role in controlling the extent to which irrelevant visual distractors are processed during visual selective attention tasks (e.g., Lavie, Hirst, De Fockert, & Viding, 2004). Recently, it has been shown that the successful selection of tactile information also depends on the availability of working memory (Dalton, Lavie, & Spence, 2009). Here, we investigate whether working memory plays a role in auditory selective attention. Participants focused their attention on short continuous bursts of white noise (targets) while attempting to ignore pulsed bursts of noise (distractors). Distractor interference in this auditory task, as measured in terms of the difference in performance between congruent and incongruent distractor trials, increased significantly under high (vs. low) load in a concurrent working-memory task. These results provide the first evidence demonstrating a causal role for working memory in reducing interference by irrelevant auditory distractors. PMID- 19557668 TI - Semantic relatedness among objects promotes the activation of multiple phonological codes during object naming. AB - In a picture-word interference experiment the authors demonstrate that a semantic categorical relation between a to-be-named target picture and a context picture promotes the phonological activation of the to-be-ignored context picture. No such phonological activation is observed if the objects are semantically unrelated. This finding gives further insight into the mechanisms that modulate the activation flow in the conceptual-lexical system during speech planning. In contrast to recent picture-picture interference studies, the results provide direct evidence that the phonological activation of a context object is dependent on its semantic processing. PMID- 19557669 TI - Random noun generation in younger and older adults. AB - We examined age-related changes of executive functions by means of random noun generation. Consistent with previous observations on random letter generation, older participants produced more prepotent responses than younger ones. In the case of random noun generation, prepotent responses are nouns of the same category as the preceding noun. In contrast to previous observations, older participants exhibited stronger repetition avoidance and a stronger tendency toward local evenness-that is, toward equal frequencies of the alternative responses even in short subsequences. These data suggest that at higher adult age inhibition of prepotent responses is impaired. In addition, strategic attentional processes of response selection are strengthened, in particular the application of a heuristic for randomness. In this sense response selection is more controlled in older than in younger adults. PMID- 19557670 TI - The value of immunoassays for metanephrines in the biochemical diagnosis of pheochromocytomas. AB - An undiagnosed pheochromocytoma may result in life-threatening consequences. The diagnosis of pheochromocytoma is based on the overproduction of catecholamines. Highly sensitive biochemical assays are essential to avoid false-negative results. Determinations of 24-h urinary epinephrine and norepinephrine levels are established diagnostic tools. However, they may be falsely negative in patients with a biochemically-silent or periodically-secreting pheochromocytoma. Metanephrines, which are metabolites of catecholamines, have been suggested as an alternative diagnostic tool. Urinary metanephrines are determined by high pressure liquid chromatography (HPLC) in an increasing number of laboratories, whereas plasma metanephrines measured by HPLC are available in specialised centres only. The different HPLC methods may be cost- and time-intensive. Immunoassays such as radio- or enzyme-immunoassays may be alternative procedures. Measurement of metanephrines instead of catecholamines by either technique improved the diagnostic accuracy for the diagnosis of pheochromocytomas. Determination of plasma free metanephrines demonstrated a higher accuracy than their urinary counterparts. The use of immunoassays may be an alternative to the laborious HPLC, although the method needs to be evaluated in more detail. PMID- 19557671 TI - Lanostanes from Phellinus igniarius and their iNOS inhibitory activities. AB - Four new lanostanol-type triterpenoids, igniarens A - D ( 1- 4), were isolated from the fruit body of Phellinus igniarius together with two known triterpenoids, and two known ergostanes. These four new compounds were identified by spectroscopic analysis as 22 R-hydroxy-24-methylene-29-norlanost-7, 9(11)-dien-3 one (1), 3alpha,22 R-dihydroxy-24-methylene-29-norlanost-7, 9(11)-diene (2), 3alpha,22 R-dihydroxy-24-methylene-29-norlanost-8-ene ( 3), and 3alpha,22 R dihydroxy-24-methylenelanost-8-ene ( 4). Their effects on NO production in lipopolysaccharide (LPS)-activated macrophages were assessed. Compounds 1- 8 inhibited NO production in activated RAW 264.7 cells to various degrees. The most potent compound 5alpha,8alpha-epidioxy-22 E-ergosta-6,22-dien-3beta-ol ( 7) significantly inhibited LPS-induced NO production in a concentration-dependent manner without affecting the cellular viability, with an IC (50) of 37.57 +/- 1.38 microM. PMID- 19557672 TI - Preface to pattern formation special issue. AB - Patterns are orders embedded in randomness; they may appear in spatial arrangements or in temporal sequences, and each element may appear identical or with variations. Patterns exist in the physical world as well as in living systems. In the biological world, patterns can range from simple to complex, forming the basic building blocks of life. When we see patterns in peacock feathers, leopard spots or zebra stripes, we are fascinated by the order, the variations and the beauty. The process that generates this ordering in the biological world has been termed Pattern Formation. Since Lewis Wolpert promoted this concept four decades ago, scientists from molecular biology, developmental biology, stem cell biology, tissue engineering, theoretical modeling and other disciplines have made remarkable progress towards understanding its underlying mechanisms. We have learned that both molecular processes and physico-chemical principles are important for biological Pattern Formation and as Guest Editors, felt that it is time to review and re-integrate our understanding of this fundamental and fantastic process. PMID- 19557674 TI - Diffusible gradients are out - an interview with Lewis Wolpert. Interviewed by Richardson, Michael K. AB - In 1969, Lewis Wolpert published a paper outlining his new concepts of "pattern formation" and "positional information". He had already published research on the mechanics of cell membranes in amoebae, and a series of classic studies of sea urchin gastrulation with Trygve Gustavson. Wolpert had presented his 1969 paper a year earlier at a Woods Hole conference, where it received a very hostile reception: "I wasnt asked back to America for many years!". But with Francis Crick lining up in support of diffusible morphogen gradients, positional information eventually became established as a guiding principle for research into biological pattern formation. It is now clear that pattern formation is much more complex than could possibly have been imagined in 1969. But Wolpert still believes in positional information, and regards intercalation during regeneration as its best supporting evidence. However, he and others doubt that diffusible morphogen gradients are a plausible mechanism: "Diffusible gradients are too messy", he says. Since his retirement, Lewis Wolpert has remained active as a theoretical biologist and continues to publish in leading journals. He has also campaigned for a greater public understanding of the stigma of depression. He was interviewed at home in London on July 26th, 2007 by Michael Richardson. PMID- 19557673 TI - Pattern formation today. AB - Patterns are orders embedded in randomness. They may appear as spatial arrangements or temporal series, and the elements may appear identical or with variations. Patterns exist in the physical world as well as in living systems. In the biological world, patterns can range from simple to complex, forming the basic building blocks of life. The process which generates this ordering in the biological world was termed pattern formation. Since Wolpert promoted this concept four decades ago, scientists from molecular biology, developmental biology, stem cell biology, tissue engineering, theoretical modeling and other disciplines have made remarkable progress towards understanding its mechanisms. It is time to review and re-integrate our understanding. Here, we explore the origin of pattern formation, how the genetic code is translated into biological form, and how complex phenotypes are selected over evolutionary time. We present four topics: Principles, Evolution, Development, and Stem Cells and Regeneration. We have interviewed several leaders in the field to gain insight into how their research and the field of pattern formation have shaped each other. We have learned that both molecular process and physico-chemical principles are important for biological pattern formation. New understanding will emerge through integration of the analytical approach of molecular-genetic manipulation and the systemic approach of model simulation. We regret that we could not include every major investigator in the field, but hope that this Special Issue of the Int. J. Dev. Biol. represents a sample of our knowledge of pattern formation today, which will help to stimulate more research on this fundamental process. PMID- 19557675 TI - Limb pattern, physical mechanisms and morphological evolution - an interview with Stuart A. Newman. Interviewed by Chuong, Cheng-Ming. AB - Stuart A. Newman grew up in New York City. He received a Bachelor of Arts from Columbia University and obtained a Ph.D. in chemical physics from the University of Chicago in 1970. He did post-doctoral studies in several institutions and disciplines with a focus on theoretical and developmental biology. He had a rich experience interacting with people like Stuart Kauffman, Arthur Winfree, Brian Goodwin, and John W. Saunders, Jr. He was also exposed to many interesting experimental models of development. These early experiences fostered his interest in biological pattern formation. He joined the State University of New York at Albany as a junior faculty member when Saunders was still there. With his physical science background, Newmans approach to limb bud patterning was refreshing. In his major Science paper in 1979, he and H.L. Frisch proposed a model showing how reaction-diffusion can produce chemical standing waves to set up limb skeletal patterns. He then used limb bud micromass cultures for further development and testing of the model. Extending earlier ideas, he developed a comprehensive framework for the role of physical mechanisms (diffusion, differential adhesion, oscillation, dynamical multistability, reaction diffusion, mechano-chemical coupling, etc.) in morphogenesis. He also applied these mechanisms to understand the origin of multicellularity and evolution of novel body plans. Here Newman reflects on his intellectual growth, and shares with us his ideas on how pattern formation works, and how generic physical mechanisms interact with genetic mechanisms to achieve the evolution and development of animal forms. PMID- 19557676 TI - Pattern formation mechanisms in reaction-diffusion systems. AB - In systems undergoing chemical reaction and diffusion, a remarkable variety of spatially structured patterns, stationary or moving, local or global, can arise, many of them reminiscent of forms and phenomena seen in living systems. Chemical systems offer the advantage that one can often control the parameters that determine the patterns formed and can thereby probe fundamental issues about pattern formation, with possible insights into biologically relevant phenomena. We present experimental examples and discuss several mechanisms by which such spatiotemporal structure may arise, classifying the mechanisms according to the type of instability that results in pattern formation. In some systems, the pattern that emerges depends not only on the chemical and physical parameters but also on the initial state of the system. Interactions between instabilities can result in particularly complex patterns. PMID- 19557677 TI - The emergence of patterning in lifes origin and evolution. AB - Three principles guide natural pattern formation in both biological and non living systems: (1) patterns form from interactions of numerous individual particles, or agents, such as sand grains, molecules, cells or organisms; (2) assemblages of agents can adopt combinatorially large numbers of different configurations; (3) observed patterns emerge through the selection of highly functional configurations. These three principles apply to numerous natural processes, including the origin of life and its subsequent evolution. The formalism of functional information, which relates the information content of a complex system to its degree of function, provides a quantitative approach to modeling the origin and evolution of patterning in living and nonliving systems. PMID- 19557678 TI - The Hox complex - an interview with Denis Duboule. Interviewed by Richardson, Michael K. AB - Denis Duboule is one of the most influential and highly-cited scientists in developmental biology. Born in Geneva in 1955, he holds dual Swiss and French nationality. His undergraduate studies in biology at the University of Geneva included research on mouse embryology. He later learned molecular techniques in the laboratory of Pierre Chambon, becoming a major player in characterising the newly-discovered vertebrate Hox genes. He helped discover their genomic clustering, realising that they had arisen by trans duplication. With Gaunt and Sharpe, he proposed that vertebrate Hox clusters might show spatial colinearity, and subsequently extended this concept to the timing of gene activation (temporal colinearity). Along with the Krumlauf laboratory, he reported the structural and functional conservation of the homeotic systems in flies and vertebrates. His lab was the first to describe nested patterns of Hox gene expression in the developing mouse limb, and later showed that digit-associated Hoxd gene expression was lacking in zebrafish paired fin development. His concept of phylotypic progression helps explain major evolutionary developmental phenomena in terms of Hox gene regulatory networks. His research helped reveal that the genital tubercle may, like the limb, be patterned by Hox genes. His lab developed targeted meiotic recombination (TAMERE), using it to make profound advances in our understanding of Hox gene regulation. Remote enhancers linked to digit patterning have been uncovered, together with a likely mechanism for colinearity. Denis lives in Geneva with his wife Brigitte Galliot, also a scientist, with their four children. PMID- 19557679 TI - Molecular tools, classic questions - an interview with Clifford Tabin. Interviewed by Richardson, Michael K. AB - Clifford J. Tabin has made pioneering contributions to several fields in biology, including retroviruses, oncogenes, developmental biology and evolution. His father, a physicist who worked in the Manhattan project, kindled his interest in science. Cliff later chose to study biology and started his research career when the world of recombinant DNA was opening up. In Robert Weinbergs lab, he constructed the Moloney leukaemia virus (MLV-tk), the first recombinant retrovirus that could be used as a eukaryotic vector. He also discovered the amino acid changes leading to the activation of Ras, the first human oncogene discovered. As an independent researcher, he began in the field of urodele limb regeneration, and described the expression of retinoic acid receptor and Hox genes in the blastema. Moving to the chick model, his was one of the labs that simultaneously cloned the first vertebrate hedgehog cognates and showed that sonic hedgehog functions as a morphogen in certain developmental contexts, in particular as an organizing activity during limb development. Comparative studies by Ann Burke in his lab showed that differences in boundaries of Hox gene expression across vertebrate phylogeny correlated with differences in skeletal morphology. The Tabin lab also discovered a genetic pathway responsible for mediating left-right asymmetry in vertebrates; helped uncover the pathways leading to dorsoventral limb patterning; made contributions to our understanding of skeletal morphogenesis and identified developmental mechanisms that might underpin the diversification of the beak in Darwins finches. Despite being a professor of genetics at Harvard, Tabin says: "I have never done a genetics experiment in my life!". This is changing with his latest project: the genetics of Mexican cavefish. I interviewed Cliff on the 3rd October, 2007, in his office at Harvard. PMID- 19557680 TI - The sudden appearance of diverse animal body plansduring the Cambrian explosion. AB - Beautifully preserved organisms from the Lower Cambrian Maotianshan Shale in central Yunnan, southern China, document the sudden appearance of diverse metazoan body plans at phylum or subphylum levels, which were either short-lived or have continued to the present day. These 530 million year old fossil representatives of living animal groups provide us with unique insight into the foundations of living animal groups at their evolutionary roots. Among these diverse animal groups, many are conservative, changing very little since the Early Cambrian. Others, especially Panarthropoda (superphylum), however, evolved rapidly, with origination of novel body plans representing different evolutionary stages one after another in a very short geological period of Early Cambrian time. These nested body plans portray a novel big picture of pararthropod evolution as a progression of step-wise changes both in the head and the appendages. The evolution of the pararthropods displays how the head/trunk boundary progressively shifted to the posterior, and how the simple annulated soft uniramous appendages progressively changed into stalked eyes in the first head appendages, into whip-like sensorial and grasping organs in the second appendage, and into jointed and biramous bipartite limbs in the post-antennal appendages. Haikouella is one of most remarkable fossils representing the origin body plan of Cristozoa, or crest animals (procraniates+craniates). The anatomy of Early Cambrian crest animals, including Haikouella and Yunnanozoon, contributes to novel understanding and discussion for the origins of the vertebrate brain, neural crest cells, branchial system and vertebrae. PMID- 19557681 TI - The cooperative genome: organisms as social contracts. AB - A predominant theme in much of evolutionary biology is that organisms are the product of relentless and precise natural selection among them, and that life is about the competition of all-against-all for success. However, developmental genetics has rapidly been revealing a very different picture of the nature of life. The organizing principles by which organisms are made are thoroughly based on complex hierarchies of molecular interactions that require multiple factors to be relentlessly cooperating with each other. Reconciling these two points of view involves changing the scale of observation, and a different understanding of evolution, in which cooperation and tolerance are more important than competition and intolerance. PMID- 19557682 TI - The evolution and maintenance of Hox gene clusters in vertebrates and the teleost specific genome duplication. AB - Hox genes are known to specify spatial identities along the anterior-posterior axis during embryogenesis. In vertebrates and most other deuterostomes, they are arranged in sets of uninterrupted clusters on chromosomes, and are in most cases expressed in a "colinear" fashion, in which genes closer to the 3-end of the Hox clusters are expressed earlier and more anteriorly and genes close to the 5-end of the clusters later and more posteriorly. In this review, we summarize the current understanding of how Hox gene clusters have been modified from basal lineages of deuterostomes to diverse taxa of vertebrates. Our parsimony reconstruction of Hox cluster architecture at various stages of vertebrate evolution highlights that the variation in Hox cluster structures among jawed vertebrates is mostly due to secondary lineage-specific gene losses and an additional genome duplication that occurred in the actinopterygian stem lineage, the teleost-specific genome duplication (TSGD). PMID- 19557683 TI - Skin, cornea and stem cells - an interview with Danielle Dhouailly. Interviewed by Chuong, Cheng-Ming. AB - Danielle Dhouailly received her Bachelor of Science degree (Biology) from Paris University. She then worked on a Ph.D. with Philippe Sengel at Grenoble University. After that, she went to Canada and the USA to work with Drs. M. Hardy, R. Sawyer and H. Sun before going back to Grenoble and starting her own laboratory. In the 1970s, she began a series of creative epithelial-mesenchymal recombination experiments among chicken feathers, mouse hairs and lizard scales, and later between rabbit cornea / mouse hairs. Through these original experiments, she elegantly demonstrated that the dermis initiates the formation of cutaneous appendages, while their type is specified by the class and regional origin of the epidermis. Subsequently she showed that the induction of an ectodermal organ, even in an adult epithelium, provokes the appearance of the related tissue stem cells. These works pioneered the concepts which are used in stem cell biology today. Her laboratory now works on the molecular mechanisms underlying these processes. Her papers are typically characterized by an initial insightful observation, followed by rigorous experiments and thoughtful discussions. They are rich with different shades of perspectives, almost like a piece of impressionist art. She loves gardening and her pets. She considers herself a good observer and hard worker driven by curiosity. Her best moments occur when she suddenly becomes enlightened as to an explanation of a basic concept when looking at experimental results or discussing ideas with colleagues. She believes that good results last forever, although interpretations can change. Her advice to young scientists is to be rigorous at the bench, to think hard, and not to be shy to speak up. The following is the story of how this young, female naturalist grew into a well-respected developmental biologist. PMID- 19557684 TI - Waves and patterning in developmental biology: vertebrate segmentation and feather bud formation as case studies. AB - In this article we will discuss the integration of developmental patterning mechanisms with waves of competency that control the ability of a homogeneous field of cells to react to pattern forming cues and generate spatially heterogeneous patterns. We base our discussion around two well known patterning events that take place in the early embryo: somitogenesis and feather bud formation. We outline mathematical models to describe each patterning mechanism, present the results of numerical simulations and discuss the validity of each model in relation to our example patterning processes. PMID- 19557686 TI - Generation of pattern and form in the developing limb. AB - The developing limb is a major model for pattern formation in vertebrate embryos. Many of the seminal discoveries of the mechanisms involved in patterning have been made using chick embryos because of the ease of manipulating their developing limbs. More recently, the molecular basis of limb pattern formation has been increasingly uncovered and now, with the availability of genomic resources, the genetic approaches available are even more powerful. Nevertheless, since the limb is ultimately built of cells, gene action must ultimately be translated into cell behaviour and a major challenge will be to integrate genetics with molecular and cellular biology. In this review, we will first outline the stages in limb development, the major interacting signalling pathways that pattern the limb and the molecules involved. We will describe fate maps of the developing limb, and discuss what is known about cellular activities including proliferation, death, adhesiveness, communication and migration during the patterning process. Finally we will explore how these cell activities produce form. PMID- 19557685 TI - Pattern formation in the Drosophila eye disc. AB - Differentiation of the Drosophila compound eye from the eye imaginal disc is a progressive process: columns of cells successively differentiate in a posterior to anterior sequence, clusters of cells form at regularly spaced intervals within each column, and individual photoreceptors differentiate in a defined order within each cluster. The progression of differentiation across the eye disc is driven by a positive autoregulatory loop of expression of the secreted molecule Hedgehog, which is temporally delayed by the intercalation of a second signal, Spitz. Hedgehog refines the spatial position at which each column initiates its differentiation by inducing secondary signals that act over different ranges to control the expression of positive and negative regulators. The position of clusters within each column is controlled by secreted inhibitory signals from clusters in the preceding column, and a single founder neuron, R8, is singled out within each cluster by Notch-mediated lateral inhibition. R8 then sequentially recruits surrounding cells to differentiate by producing a short-range signal, Spitz, which induces a secondary short-range signal, Delta. Intrinsic transcription factors act in combination with these two signals to produce cell type diversity within the ommatidium. The Hedgehog and Spitz signals are transported along the photoreceptor axons and reused within the brain as long range and local cues to trigger the differentiation and assembly of target neurons. PMID- 19557688 TI - Regeneration and pattern formation - an interview with Susan Bryant. Interviewed by Richardson, Michael K and Chuong, Cheng-Ming. AB - Susan Bryant is one of the leading researchers in regeneration and pattern formation. Born in England in 1943, she studied biology at Kings College, London (UK). After a Ph.D. with Angus Bellairs on caudal autotomy and regeneration in lizards, she researched urodele regeneration in Marcus Singer's lab at Case Western Reserve University. Then, at the University of California, Irvine, she adopted the axolotl as a research model for limb regeneration and pattern formation. Her work supported models involving the intercalation of positional values in a polar coordinate system. Fibroblasts, often regarded as "junk" cells, are seen by Susan Bryant as central to patterning. She argues that fibroblasts express positional values needed for regeneration. She also argues that vertebrate species capable of regeneration have evolved steps to plug back into developmental programmes. Susan Bryant thinks that regeneration is essential for a full understanding of development, and believes that developmental biology has suffered though not embracing regeneration. She also believes that deeper knowledge of pattern formation will bring advances in emerging field of tissue engineering. Since 2000, she has served as Dean of Biological Sciences and more recently, as Vice Chancellor for Research, at UC Irvine (USA). She is an advocate of equal opportunities for women and other under-represented groups in academia. She lives in California with husband David Gardiner, her scientific partner for over 20 years. They have two children. We interviewed Susan Bryant in her office in Irvine on October 5th, 2007. PMID- 19557689 TI - Zebrafish development and regeneration: new tools for biomedical research. AB - Basic research in pattern formation is concerned with the generation of phenotypes and tissues. It can therefore lead to new tools for medical research. These include phenotypic screening assays, applications in tissue engineering, as well as general advances in biomedical knowledge. Our aim here is to discuss this emerging field with special reference to tools based on zebrafish developmental biology. We describe phenotypic screening assays being developed in our own and other labs. Our assays involve: (i) systemic or local administration of a test compound or drug to zebrafish in vivo; (ii) the subsequent detection or "readout" of a defined phenotypic change. A positive readout may result from binding of the test compound to a molecular target involved in a developmental pathway. We present preliminary data on assays for compounds that modulate skeletal patterning, bone turnover, immune responses, inflammation and early-life stress. The assays use live zebrafish embryos and larvae as well as adult fish undergoing caudal fin regeneration. We describe proof-of-concept studies on the localised targeting of compounds into regeneration blastemas using microcarriers. Zebrafish are cheaper to maintain than rodents, produce large numbers of transparent eggs, and some zebrafish assays could be scaled-up into medium and high throughput screens. However, advances in automation and imaging are required. Zebrafish cannot replace mammalian models in the drug development pipeline. Nevertheless, they can provide a cost-effective bridge between cell-based assays and mammalian whole-organism models. PMID- 19557687 TI - Reptile scale paradigm: Evo-Devo, pattern formation and regeneration. AB - The purpose of this perspective is to highlight the merit of the reptile integument as an experimental model. Reptiles represent the first amniotes. From stem reptiles, extant reptiles, birds and mammals have evolved. Mammal hairs and feathers evolved from Therapsid and Sauropsid reptiles, respectively. The early reptilian integument had to adapt to the challenges of terrestrial life, developing a multi-layered stratum corneum capable of barrier function and ultraviolet protection. For better mechanical protection, diverse reptilian scale types have evolved. The evolution of endothermy has driven the convergent evolution of hair and feather follicles: both form multiple localized growth units with stem cells and transient amplifying cells protected in the proximal follicle. This topological arrangement allows them to elongate, molt and regenerate without structural constraints. Another unique feature of reptile skin is the exquisite arrangement of scales and pigment patterns, making them testable models for mechanisms of pattern formation. Since they face the constant threat of damage on land, different strategies were developed to accommodate skin homeostasis and regeneration. Temporally, they can be under continuous renewal or sloughing cycles. Spatially, they can be diffuse or form discrete localized growth units (follicles). To understand how gene regulatory networks evolved to produce increasingly complex ectodermal organs, we have to study how prototypic scale-forming pathways in reptiles are modulated to produce appendage novelties. Despite the fact that there are numerous studies of reptile scales, molecular analyses have lagged behind. Here, we underscore how further development of this novel experimental model will be valuable in filling the gaps of our understanding of the Evo-Devo of amniote integuments. PMID- 19557690 TI - How animals get their skin patterns: fish pigment pattern as a live Turing wave. AB - It is more than fifty years since Alan Turing first presented the reaction diffusion (RD) model, to account for the mechanism of biological pattern formation. In the paper entitled "The chemical basis of morphogenesis", Turing concluded that spatial patterns autonomously made in the embryo are generated as the stationary wave of the chemical (cellular) reactions. Although this novel idea was paid little attention by experimental biologists, recent experimental data are suggesting that the RD mechanism really functions in some of the course of animal development. Among the phenomena in which involvement of the RD mechanism is suspected, the striped pigment pattern of zebrafish has been highlighted as an ideal model system for the following reasons: the stationary wave made by the RD mechanism stays alive and can be observed only in the fish skin; and in zebrafish, we can utilize genomic information and molecular genetic techniques to clarify the molecular basis of pattern formation. In this review, we summarize recent progresses in the study of zebrafish pigment pattern formation that is uncovering how the RD wave is made and maintained in the skin. PMID- 19557691 TI - Regenerative patterning in Swarm Robots: mutual benefits of research in robotics and stem cell biology. AB - This paper presents a novel perspective of Robotic Stem Cells (RSCs), defined as the basic non-biological elements with stem cell like properties that can self reorganize to repair damage to their swarming organization. Self here means that the elements can autonomously decide and execute their actions without requiring any preset triggers, commands, or help from external sources. We develop this concept for two purposes. One is to develop a new theory for self-organization and self-assembly of multi-robots systems that can detect and recover from unforeseen errors or attacks. This self-healing and self-regeneration is used to minimize the compromise of overall function for the robot team. The other is to decipher the basic algorithms of regenerative behaviors in multi-cellular animal models, so that we can understand the fundamental principles used in the regeneration of biological systems. RSCs are envisioned to be basic building elements for future systems that are capable of self-organization, self-assembly, self-healing and self-regeneration. We first discuss the essential features of biological stem cells for such a purpose, and then propose the functional requirements of robotic stem cells with properties equivalent to gene controller, program selector and executor. We show that RSCs are a novel robotic model for scalable self-organization and self-healing in computer simulations and physical implementation. As our understanding of stem cells advances, we expect that future robots will be more versatile, resilient and complex, and such new robotic systems may also demand and inspire new knowledge from stem cell biology and related fields, such as artificial intelligence and tissue engineering. PMID- 19557692 TI - [Psychometric attributes of the Scales for Outcomes in Parkinson's Disease Psychosocial (SCOPA-PS): validation in Spain and review]. AB - AIMS: To evaluate the psychometric attributes of the Scales for Outcomes in Parkinson's Disease-Psychosocial (SCOPA-PS) in Spain and to compare them with previous studies. PATIENTS AND METHODS: We performed a multi-centre, cross sectional study of 387 patients with Parkinson's disease (PD), 70% of whom were in Hoehn and Yahr (HY) stages 2 or 3, with a mean age of 65.8 +/- 11.1 years and 8.1 +/- 6 years' progression. The following measures were applied: SCOPA-Motor, SCOPA-Cognition, modified Parkinson's Psychosis Rating Scale, Clinical Impression of Severity Index for Parkinson's Disease (CISI-PD), Cumulative Illness Rating Scale-Geriatrics, SCOPA-Autonomic, SCOPA-Sleep, Hospital Anxiety and Depression Scale (HADS), Fatigue and Pain Visual Analogue Scales, EQ-5D and SCOPA-PS. Acceptability, internal consistence, dimensionality, construct validity and precision of the SCOPA-PS were analysed. RESULTS: The SCOPA-PS summary index displayed no ceiling or floor effect. Internal consistence was satisfactory (alpha = 0.85; item-total correlation => 0.39). Two factors were identified (53.5% of the variance). The SCOPA-PS was highly correlated (r(S) => 0.5) with the HADS, SCOPA-Motor, SCOPA-Automatic and EQ-5D index, and moderately so (r(S) = 0.35-0.49) with CISI-PD, fatigue and HY. The SCOPA-PS discriminated significantly between patients grouped according to HY stages, levels of severity of the CISI PD, age groups and length of time with PD. The standard error of the measure was 7.24 +/- 18.7. As a whole, these findings are in agreement with previous studies. CONCLUSIONS: The SCOPA-PS is a scale with a satisfactory degree of acceptability, and it is consistent, valid and precise for evaluating the psychosocial impact of PD. PMID- 19557693 TI - [Predisposition to alcohol and drug consumption in schizophrenia-vulnerable people]. AB - INTRODUCTION: A large amount of current schizophrenia research has been centered on the understanding of its etiological mechanisms and the detection of vulnerability markers in people at risk. This vulnerability called schizotypy can be identified in people not affected by the illness at the clinical level. AIM: To check if the schizotypic personality disorder as a vulnerability marker of the disorders in the schizophrenic spectrum predicts the presence of psychopathologic symptoms and alcohol and drugs intake. SUBJECTS AND METHODS: From a population of 442 university students tested with the Schizotypy Personality Questionnaire (SPQ), it was selected a sample including people scoring on the 20% superior and inferior for the characteristic factors of schizophrenia corresponding to positive symptoms (cognitive-perceptual), negative symptoms (interpersonal) and thought disorder (disorganized). Furthermore, it was evaluated the presence of psychopathological problems and symptoms. Also, the participants gave information about alcohol and drugs intake as a passive coping strategy with stress. RESULTS: Compared with the low scored, subjects with high scores in the schizotypic personality disorder showed a significant increase in the presence of psychopathological problems and symptoms and a higher alcohol and drugs intake. That occurs mainly when those scores are found in symptoms associated to thought disorder and negative symptoms as lack of interest in social activities and emotional flattening. CONCLUSIONS: The schizotypic personality disorder, in accordance with its role as vulnerability factor, seems to co-occur with a higher volume of somatic and psychopathologic symptoms, and alcohol and drugs intake. PMID- 19557694 TI - [Preoperative embolisation with absorbable gelatine sponge in intracranial meningiomas]. AB - AIMS: The purpose of this study was to evaluate the benefits of preoperative embolisation with an absorbable gelatine sponge in selected intracranial meningiomas. PATIENTS AND METHODS: We performed a retrospective study of a series of 33 patients who had undergone surgery for meningiomas with a diameter of over 4 cm between the years 2000 and 2007. Two groups were analysed: group A (n = 16) with preoperative embolisation and group B (n = 17) without it. Eligibility criteria for embolisation were: exclusive or predominant irrigation through the external carotid artery, high tumour flow and pronounced vascularisation through the pial branches. The location of the lesion was evaluated preoperatively; blood losses, number of units of blood transfused, surgery time and surgeon's opinion were evaluated intraoperatively. RESULTS: A statistically significant difference was found between the group of embolised patients and those who had not been embolised as far as surgery time (217.5 +/- 69.61 versus 291.76 +/- 56.94 min; p =< 0.002) and blood loss (613.75 +/- 231.42 versus 987.65 +/- 206.68 mL; p => 0.001) were concerned. A positive coloration (r = 0.568; p = 0.001) was found between surgery time and blood loss. No statistically significant relation was observed between age and the number of units of blood transfused. Embolisation was considered to be beneficial by 75% of surgeons. CONCLUSIONS: Preoperative embolisation with an absorbable gelatine sponge in patients with intracranial meningiomas with a diameter above 4 cm and exclusive or predominant irrigation by the external carotid artery is effective and safe; it also reduces intraoperative bleeding and surgery time. PMID- 19557695 TI - [Headache in idiopathic intracranial hypertension]. AB - INTRODUCTION: Idiopathic intracranial hypertension (IICH) is characterized by a rising in intracranial hypertension without evidence of intracranial expansive disease. Diagnostic criteria of headache related to IICH are described in the International Classification of Headache Disorders. In clinical practice, however, headache related to IICH may be heterogeneous. PATIENTS AND METHODS: Clinical charts of patients with IICH admitted to the Hospital Universitario de Gran Canaria Dr. Negrin between 1990 and 2007 were retrospectively reviewed. RESULTS: Fifty-five patients were included, with a ratio female/male of 8.1:1. Headache was present in 85.4%. The headache was continuous in 63.8% of patients, and diffuse in 51%. CONCLUSION: Headache related to IICH is heterogeneous, and may mimic primary headache, so a high level of suspicion is needed to avoid diagnostic delay. PMID- 19557696 TI - [Dejerine-Roussy syndrome of an ischaemic origin in an adolescent with patent foramen ovale]. AB - INTRODUCTION: Dejerine-Roussy syndrome, or thalamic syndrome, is characterised by transient mild hemiparesis, hemichoreoathetosis, hemihypoesthesia, hyperalgesia, allodynia and hemiataxia with astereognosia that varies in intensity, and it appears in the presence of lesions in the posterior nuclei of the thalamus. It can be produced by strategic cerebral infarction, reported in elderly patients with vascular risk factors. Patent foramen ovale has been suggested as a risk factor for ischaemic stroke in young people, especially when associated to aneurysm of the auricular septum and above all to a procoagulating status. CASE REPORT: An 18-year-old male with a family history of Behcet's disease, who presented right-side thalamic and hippocampal cerebral infarction; following an exhaustive study, patent foramen ovale with septal aneurysm was found as the only risk factor. At that time he did not satisfy criteria for Behcet's disease, and thorough systemic screening did not reveal direct or indirect signs of venous thrombosis. Percutaneous closure of the foramen was performed. CONCLUSIONS: This is the first reported case of Dejerine-Roussy syndrome as a manifestation of cryptogenic cerebral infarction associated to patent foramen ovale in an adolescent. Taken as a whole, the clinical and complementary data enable us to reconstruct the pathophysiological sequence that position foramen ovale with an associated septal 'aneurysm' as the only detectable risk factor, which, when linked to the stress of the patient and the family, triggered its early closure. PMID- 19557697 TI - [The role of the neuromodulators in the preventive treatment of migraine]. AB - INTRODUCTION: More than one quarter of patients who consult due to migraine need preventive treatment. At least half of these patients do not respond or cannot tolerate classical preventatives: beta-blockers, flunarizine or amitriptyline. AIM: To analyse the role of new antiepileptics (neuromodulators) in migraine prevention. DEVELOPMENT: Critical review of clinical trials carried out with antiepileptics for the treatment of migraine. Topiramate has demonstrated clear efficacy in the prevention of migraine with and without aura, including chronic migraine with and without analgesic overuse, although it is not tolerated by one fifth of patients. Valproic acid also has a demonstrated efficacy in the preventive treatment of migraine with and without aura, but its tolerability profile and the possibility of serious adverse events oblige us to prescribe this neuromodulator for refractory patients. Gabapentin shows a mild efficacy, clearly lower than that of topiramate and valproate. Lamotrigine is the treatment of choice in the prevention of migraine aura, but is not efficacious in the treatment of headache. Levetiracetam has shown some efficacy in open studies in migraine prevention, both in adults and children. Zonisamide has demonstrated good efficacy and tolerability in patients refractory to topiramate and shows a prolonged half life, which improves compliance. The remaining antipileptics have not shown efficacy. CONCLUSIONS: Topiramate and valproic acid have demonstrated efficacy in migraine prevention. Lamotrigine is efficacious for the treatment of migraine aura. Even though new studies are necessary, zonisamide could be a good option for the preventive treatment of migraine. PMID- 19557698 TI - [Reconsiderations in the treatment of Parkinson's disease with levodopa: some pharmacodynamic evidence]. AB - AIM: To review the current literature on the use of levodopa in the treatment of Parkinson's disease (PD), with an emphasis on pharmacodynamical aspects. DEVELOPMENT: Levodopa has been used successfully for treatment of PD since its discovery, being still the drug of choice. It has shown decreasing motor disability and reduce mortality in PD patients, however, still maintains long term problems, particularly in relation to the motor response, the onset of dyskinesias and apparent neurotoxicity. The pharmacodynamical evidence indicates that pharmacological effects such as short-term response (SDR) and the long-term response (LDR) are an integral part of the therapeutic response to levodopa. Both responses have pharmacodynamical changes in time and its effects are expressed in terms of amplitude, duration and onset latency of pharmacological action. Investigations in this area, show that LDR would the expression of the physiological release of dopamine, and the treatment of PD should be based on LDR and its pharmacological characteristics. CONCLUSIONS: The pharmacodynamics of levodopa is complex. However, the available evidence suggests that a therapeutic strategy based on the use of the LDR is better than those based on the control of the SDR. The knowledge of the pharmacological characteristics of levodopa is central to the therapeutic decision, which should be effective in reducing symptoms and have minimal side effects. PMID- 19557700 TI - [Horner syndrome secondary to a central venous line]. PMID- 19557702 TI - [Transient neurological deficit and chronic subdural haematoma]. PMID- 19557699 TI - [Consensus clinical practice guidelines of the Andalusian Epilepsy Society on prescribing generic antiepileptic drugs]. AB - INTRODUCTION: Pharmaceutical spending in Spain accounts for 1.2-1.4% of the gross domestic product and is increasing by 5-12% per year. One of the measures adopted by the government to cut this spending is the possible substitution of original prescribed drugs by generics. In the case of antiepileptic drugs (AED), which are characterised by a scant therapeutic margin, these steps have sparked a scientific debate about their repercussion on the control of epileptic patients. We propose to draw up a set of implicit evidence-based consensus practice guidelines concerning issues related with this topic. DEVELOPMENT: A selective search for quality scientific information on the subject was conducted on PubMed Medline, Tripdatabase and the Biblioteca Cochrane Plus. The selected references were analysed and discussed by the authors, and the recommendations deriving from them were collected. A total of 21 primary documents and 16 practice guidelines, protocols or experts' recommendations were identified. Our recommendations were explicitly included at the end of the text. CONCLUSIONS: The Andalusian Epilepsy Society makes the following recommendations: 1) not replacing an innovative AED by its generic in a controlled patient; 2) beginning treatment with a generic AED in monotherapy or in association is acceptable; 3) not exchanging generic AED from different pharmaceutical companies; 4) explaining to the patient the rules governing the authorization of generics and the importance of avoiding exchanges between different generic AED; and 5) if there is some worsening of the clinical condition or side effects appear following the introduction of a generic, the causes must be investigated and communicated to the bodies responsible for pharmacovigilance. PMID- 19557703 TI - [Electromyographic pattern in patients and asymptomatic carriers of the SCA2 mutation]. PMID- 19557704 TI - [A clinical approach to cranial neuralgias]. PMID- 19557705 TI - Methodological considerations for observational studies of acute kidney injury using existing data sources. AB - Acute kidney injury (AKI), as defined by small, often reversible changes in kidney function, has recently been recognized as an important complication in hospitalized patients, and has been consistently associated with prolonged hospital length of stay, increased associated costs and short-term mortality. Research studies on the epidemiology of AKI must address a number of unique methodological challenges, which have the potential to impact study results and validity. This review explores several methodological issues relevant to the design and conduct of observational studies that employ preexisting laboratory, administrative or research databases and that examine AKI as an outcome or an exposure. We discuss how methodological decisions may affect study results, particularly as they relate to selection bias, misclassification and confounding. Highlighting these areas may facilitate the design of studies of high methodological rigor that advance our understanding of AKI. PMID- 19557706 TI - Renin, prorenin and the kidney: a new chapter in an old saga. AB - The binding of prorenin to the (pro)renin receptor triggers 2 major pathways: a nonproteolytic conformational change in prorenin to its active form (angiotensin II-dependent pathway) and an intracellular pathway via the (pro)renin receptor itself (angiotensin II-independent pathway). In diabetic animals, an increased plasma prorenin level not only causes the generation of angiotensin II via the angiotensin II-dependent pathway, it also stimulates the transliteration receptors own intracellular signaling pathway in a manner that is independent of the generated angiotensin II. Thus, the administration of a "handle" region peptide (HRP), which acts as a decoy peptide and competitively inhibits the binding of prorenin to the (pro)renin receptor, has a beneficial effect in the kidneys of diabetic animals with low plasma renin levels. However, the benefits of HRP are slightly reduced in animal models of essential hypertension with relatively high plasma renin levels, and these benefits disappear altogether in animal models of hypertension with extremely high plasma renin levels. Thus, in the kidneys of animal models of diabetes and/or hypertension, both renin and prorenin competitively bind to the (pro)renin receptor and contribute to the pathophysiology of nephropathy. Consequently, renin, prorenin and the (pro) renin receptor may be important therapeutic targets for the prevention and regression of nephropathy in patients with diabetes and/or hypertension. PMID- 19557707 TI - Xenobiotic kidney organogenesis: a new avenue for renal transplantation. AB - Currently many efforts are being made to apply regenerative medicine to clinical renal diseases. It has been suggested that some renal diseases which maintain renal structure can be treated by infusion of stem cells isolated from the bone marrow or adult kidney. However such cell-based therapy cannot be applied to the treatment of chronic renal disease, in which renal structure, including the kidney scaffold, is totally disrupted. Therefore, absolute kidney regeneration is needed to rebuild a whole functional kidney de novo and eliminate the requirement for dialysis. However, due to the anatomical complexity of the kidney and the need for communication between each cell to fulfill renal function, the kidney has been labeled as the most difficult organ to regenerate. Only a small number of groups are investigating the potential for reconstructing an organized and functional kidney structure, and, among them, we are using the developing xenoembryo as an organ factory for this purpose. Here we review the challenges faced in developing a whole functional kidney de novo and discuss the obstacles which must be overcome before clinical use is possible. PMID- 19557708 TI - Sleep disturbances and sleep apnea in patients on chronic peritoneal dialysis. AB - Sleep disturbances in chronic peritoneal dialysis (PD) patients are increasingly widely recognized. These include changes in sleep architecture, sleep apnea syndrome, restless legs syndrome, periodic limb movement disorder and excessive daytime sleepiness. Some of these entities will be addressed in this review. The study tools for detecting sleep-related disorders in PD patients range from self reported questionnaires (subjective) to standard overnight polysomnographic measurements (objective). Both these tools identify a high prevalence (over 50%) of sleep-related disorders among dialysis patients. Among these, sleep apnea syndrome is one of the most widely reported sleep-related disorders in PD. Sleep apnea in PD may be attributed to chronic fluid overload and uremia, leading to both obstructive and central forms of apneas. The exact underlying pathophysiology, however, is complex and likely involves a combination of multiple factors. There are preliminary data that nocturnal PD may be advantageous over conventional continuous ambulatory peritoneal dialysis in correcting sleep apnea associated with PD. Randomized studies are needed for confirmation. PMID- 19557709 TI - There is a choice for immunosuppressive drug nephrotoxicity: Is it time to change? AB - The central issue in organ transplantation remains suppression of allograft rejection. Thus, development of immunosuppressive drugs is the key to successful allograft function. New immunosuppressive drugs were introduced on the basis of their ability to reduce the incidence of acute rejection and to demonstrate shortterm outcomes at least equivalent to those achieved with the use of established immunosuppressive therapy. Although short-term renal allograft survival has improved since the introduction of calcineurin inhibitors (CNIs), long-term renal allograft survival remains a major concern, with chronic allograft nephropathy (CAN) being the principal cause of renal allograft loss after the first year. CAN has traditionally been viewed as the result of repeated low-grade immune responses directed against allogeneic tissue, but recent evidence indicates that nonimmunological or alloantigen-independent factors also contribute to its pathogenesis. CNI nephrotoxicity occurs soon after initiation of therapy, is more clearly dose-dependent. This scenario presents a clear need for new strategies that produce adequate immunosuppression to prevent acute rejection and simultaneously reduce adverse effects associated with CNI-related therapies. To obtain significant long-term improvement in renal allograft outcomes, it may be necessary to adopt new immunosuppressive regimens that rely less on CNIs. PMID- 19557710 TI - Contrast-induced nephropathy and its prevention: What do we really know from evidence-based findings? AB - INTRODUCTION: Contrast-induced acute kidney injury, also referred to as contrast induced nephropathy (CIN), is a potentially serious renal complication associated with the use of iodinated contrast media (CM) in patients at risk. With the dramatic growth in contrast-enhanced imaging services worldwide, including procedures involving exposure to iodinated CM, efforts to reduce the occurrence of CIN have received considerable attention in recent years. To date, these efforts have met with little success since the 12% prevalence of CIN today remains unchanged from 2 decades ago. METHODS: We conducted a systematic literature review of the most recent evidence available from published reports of contemporary (2000-2008) prospective, randomized, controlled trials that have investigated CIN either by comparing CM or by comparing preventive strategies. The objective was to critically review the findings in light of several aspects of study design and then to establish a set of parameters for consideration in the planning of future CIN trials so as to optimize the strength of evidence obtained. RESULTS: Whether future CIN trials are investigating comparative CM nephrotoxicity or dealing with prophylactic strategies for risk reduction, the complexities that must be addressed include a standardized definition of CIN, appropriate timing of SCr measurements with timing standardized for all subjects in a given study population, awareness of study population risk profile, hydration protocols, and pharmacological prophylactic strategies. CONCLUSIONS: Large, well-designed trials (ideally with hard clinical outcome measures) that consider all the complexities involved in CIN and its prevention are needed before the clinical community has the evidence-based direction required for optimized patient care. PMID- 19557711 TI - Evolution of protein-bound uraemic solutes during predilution haemofiltration. AB - BACKGROUND: Protein-bound uraemic toxins provoke multiple biological changes involved in uraemia. Few if any dialytic strategies remove these compounds. METHODS: In this post hoc analysis of remnant samples from a randomised controlled trial, we evaluate whether predilution haemofiltration (HF) decreases the pretreatment concentration of protein- bound uraemic solutes. Patients treated with low-flux haemodialysis (HD) were enrolled into a group continuing this strategy (group A, n=8) over 6 months, whereas group B (n=12) was switched to predilution online HF. Blood was sampled at baseline and after 6 months to determine total and free concentration and percentage binding of indoxyl sulfate (IS), indole-3-acetic acid (IAA), hippuric acid (HA), p-cresol (PC) and 3-carboxy 4-methyl-5-propyl-2-furanpropionic acid (CMPF). RESULTS: Comparing concentrations at start versus 6 months of treatment by paired analysis, HD had no impact. In contrast, at the end of the HF period, we found a decrease in total and free PC, free IAA and total CMPF. In addition, the percentage protein binding of IAA increased significantly. However, unpaired analysis revealed no statistical difference between HD and HF, both at baseline and after 6 months of treatment for all compounds. CONCLUSIONS: Paired analysis showed a beneficial impact of predilution online HF for several proteinbound uraemic solutes. Unpaired analysis, however, showed no statistical difference. PMID- 19557712 TI - Clinical features of hemodialysis patients with intimal versus medial vascular calcifications. AB - BACKGROUND: Vascular calcifications (VCs) contribute to the massive mortality in hemodialysis (HD) patients. We aimed to identify prevalence and risk factors for arterial medial calcifications (AMCs) versus intimal calcifications (AICs) in a single-center HD population. METHODS: This cross-sectional study included 134 patients, mean age 56.9 +/- 9.7 years, on HD for 8.2 +/- 5.0 years. VCs were scored based on plain radiographs and ultrasonography of the common carotid arteries. RESULTS: Patients were categorized into groups I (13% without VC), II (10% with an AMC pattern), III (24% with an AIC pattern) and IV (53% with a mixed pattern). AIC and mixed patterns were associated with older age (p=0.006 and p=0.004, respectively), and mixed pattern with longer dialysis vintage (p=0.001). Pulse pressure was significantly higher in patients from group III than group IV, and intima-media thickness (IMT) was higher in both groups with AIC. By multivariate analysis, risk factors for any VC were high serum Ca, phosphate, CaxP product, low total protein, high body mass index (BMI), systolic and diastolic blood pressure, IMT and history of smoking. Elevated calcium and/or phosphate predicted an AMC pattern, and high calcium, BMI and IMT an AIC pattern. Finally, high IMT, systolic blood pressure, BMI and older age were predictors of a mixed pattern. CONCLUSION: We observed a very high prevalence of VC, mostly with a mixed AIC+AMC pattern. Apart from well-known risk factors, the data stress the importance of smoking, an under-recognized cause of AMC, and systolic blood pressure for AIC+AMC. PMID- 19557713 TI - Cardiac magnetic resonance imaging in hemodialysis patients. AB - BACKGROUND: The anatomy of left ventricular hypertrophy (LVH) in dialysis patients was studied with magnetic resonance imaging (MRI). Potential benefits of spironolactone were examined in a subset of patients. METHODS: This was a prospective case series of 30 hemodialysis patients in whom cardiac MRI was performed. Repeat MRI was done in a subset of 13 patients after 9 months of daily oral spironolactone 25 mg. RESULTS: Subjects exhibited a characteristic cardiac morphology, distinct from both concentric LVH (cLVH) and eccentric LVH (eLVH). Compared with normal controls, LV mass index was increased, end-diastolic volume index was increased, and ejection fraction was reduced, but sphericity indices did not differ. No significant changes were seen after spironolactone. CONCLUSIONS: LVH in dialysis patients has a unique MRI appearance which is distinct from either cLVH or eLVH, with profound LV thickening and moderate dilation of the ventricular cavity. A 9-month course of spironolactone therapy did not affect morphology. PMID- 19557715 TI - Occult HBV infection in hemodialysis setting is marked by presence of isolated antibodies to HBcAg and HCV. AB - BACKGROUND: Hepatitis B virus (HBV) infections are a matter of concern in hemodialysis units; occult HBV infections (serum HBsAg negative but HBV DNA positive) were demonstrated in this setting, and this involves further concerns regarding possible transmission and pathogenic consequences. This study aimed to investigate the prevalence and correlates of occult HBV infection in a group of patients with reference to a single hemodialysis unit in southeastern Italy. METHODS: We analyzed HBV serology and DNA (using a qualitative nested PCR) in 128 HBsAg-negative hemodialysis patients, and correlated the results obtained, with sex, age, hemodialysis duration and HCV seropositivity. RESULTS: As a whole, occult HBV infection was demonstrated in 34/128 patients (26.6%); HBV DNA detection was more frequent when anti-HBcAg antibodies were detected in isolation (72%) than when associated with anti-HBsAg antibodies (31%). Among HCV seropositive patients, occult HBV infection was observed in 66%, and among these as many as 14/15 patients (93%) who were HCV+/anti-HBcAg+ had serum HBV DNA detectable. On multivariate analysis, HCV seropositivity and the presence of anti HBs were still respectively correlated to the presence and absence of occult HBV infection. CONCLUSIONS: Occult HBV infection is frequent among hemodialysis patients in our geographical area, particularly correlated to the presence of isolated anti-HBcAg and anti-HCV antibodies. Thus, the presence of isolated anti HBcAg should prompt the clinician to evaluate a possible occult HBV infection especially if anti-HCV antibodies are also detectable; this condition, in fact, seems to strongly predict the detection of HBV DNA. PMID- 19557714 TI - Decreased GFR estimated by MDRD or Cockcroft-Gault equation predicts incident CVD: the strong heart study. AB - BACKGROUND: Kidney function, expressed as glomerular filtration rate (GFR), is commonly estimated from serum creatinine (Scr) and, when decreased, may serve as a nonclassical risk factor for incident cardiovascular disease (CVD). The ability of estimated GFR (eGFR) to predict CVD events during 5-10 years of follow-up is assessed using data from the Strong Heart Study (SHS), a large cohort with a high prevalence of diabetes. METHODS: eGFRs were calculated with the abbreviated Modification of Diet in Renal Disease study (MDRD) and the Cockcroft-Gault (CG) equations. These estimates were compared in participants with normal and abnormal Scr. The association between eGFR and incident CVD was assessed. RESULTS: More subjects were labeled as having low eGFR (<60 ml/min per 1.73 m2) by the MDRD or CG equation, than by Scr alone. When Scr was in the normal range, both equations labeled similar numbers of participants as having low eGFRs, although concordance between the equations was poor. However, when Scr was elevated, the MDRD equation labeled more subjects as having low eGFR. Persons with low eGFR had increased risk of CVD. CONCLUSIONS: The MDRD and CG equations labeled more participants as having decreased GFR than did Scr alone. Decreased eGFR was predictive of CVD in this American Indian population with a high prevalence of obesity and type 2 diabetes mellitus. PMID- 19557716 TI - Fractional urinary excretion of IgG is the most powerful predictor of renoprotection by ACE inhibitors in IgA nephropathy. AB - BACKGROUND: Several aspects of renoprotection by angiotensin-converting enzyme inhibitors (ACEi) in IgA nephropathy (IgAN) are poorly defined: factors affecting responsiveness, role of proteinuria components and histological lesions, and criteria to identify patients who may benefit from ACEi. METHODS: In an observational study of 140 IgAN patients (follow up 62 +/- 36 months), 73 untreated and 67 ACEitreated for 53 +/- 28 months, 9 baseline risk factors (RFs) (blood pressure, serum creatinine, proteinuria/day, fractional excretion of IgG [FEIgG] and alpha1-microglobulin, global and segmental [SS] glomerular sclerosis, tubulointerstitial damage and arteriolar hyalinosis [AH] score), each divided into 2 subgroups according to a cutoff with the highest sensitivity and specificity for progression, were evaluated for ability to predict renoprotection. Primary end point: end-stage renal disease (ESRD) and doubling of serum creatinine (sCr); secondary end point: increase >or=25% of sCr with last sCr >or=1.58 mg/dL; total progression: sum of end points. RESULTS: Patients with RFs below cutoffs did not benefit from ACEi. All clinical and proteinuric and 2 histological RFs (SS, AH score) with values above cutoffs showed significant reduction of progression in ACEitreated vs. untreated patients; FEIgG showed the highest prediction of renoprotection: ESRD/sCrx2: 20% vs. 62% (p=0.0004); total progression: 40% vs. 85% (p=0.0003). By multivariate analysis, independent predictors of progression were FEIgG, sCr and no ACEi treatment. Proteinuria reduction from -100% to -30%, spontaneous or after ACEi treatment, did not affect progression in treated vs. untreated patients (19% vs. 13%, p=0.85). Patients with proteinuria increased or reduced <30% showed a reduction of total progression if ACEi-treated (15% vs. 77%, p=0.0002). Presence of 1 clinical or proteinuric RF above the cutoff may be a criterion to identify patients who may benefit from ACEi. CONCLUSIONS: Renoprotection by ACEi is a multifactorial phenomenon: the best predictor of renoprotection is FEIgG, a marker of disruption of glomerular barrier to proteins; renoprotection depends not only on ability to reduce proteinuria, but probably also on antiinflammatory and antifibrotic activity. PMID- 19557717 TI - Storage at -80 degrees C decreases the concentration of HPLC-detected urinary albumin: possible mechanisms and implications. AB - BACKGROUND: Urinary albumin is now measured by high-performance liquid chromatography (HPLC) which also detects albumin missed by traditional immunochemical methods. A predictive effect of HPLC-detected albuminuria on mortality has just been reported in the AusDiab study, measuring albuminuria with HPLC after 7 years of -80 degrees C storage. However, there are already some data suggesting that HPLC-detected albuminuria is affected by -80 degrees C storage. We aimed to measure changes in HPLC-detected albuminuria after 2.5 years and find the factors which may be responsible for this alteration. METHODS: Urinary albumin was measured by the US Food and Drug Administration approved HPLC Accumin kit. Total free sulfhydryl groups (TFSG) of urine samples were measured by Ellman's reagent. RESULTS: We found a significant 24% average decrease in HPLC detected albuminuria and a correlation between the magnitude of decrease and urinary pH. We found a correlation between changes of urinary albumin dimeric to monomeric ratio of stored urine and pH; however, only changes of monomeric form were found to be significant. A correlation was also found between the TFSG of fresh urine samples and pH. Less TFSG could be detected, and a correlation between TFSG and pH was absent in stored urine. CONCLUSIONS: We conclude that measurement of albuminuria by HPLC in long-term -80 degrees stored urine gives unreliable results. Decrease of HPLC-detected albuminuria is pH-dependent and may be due to the reducing capacity of urine. Prospective studies need to decide whether the predictive properties of HPLC-detected albuminuria decrease during longterm storage. PMID- 19557719 TI - Multicystic nephroma: a rare entity. AB - Multicystic nephroma is a rare renal pathology, which is characterized usually by a unilateral renal mass with multiple thin-walled cysts divided by septa. We report the case of a boy aged 2 years, who was diagnosed in utero with an abdominal cystic lesion during routine antenatal ultrasonography examination. The patient underwent a nephrectomy, and diagnosis of cystic nephroma was confirmed by histology. PMID- 19557720 TI - Spontaneous coronary artery dissection: One more extrarenal manifestation of autosomal dominant polycystic kidney disease? AB - Autosomal dominant polycystic kidney disease (ADPKD) may present with a variety of extrarenal manifestations. Only 1 case of spontaneous coronary artery dissection has been reported so far in ADPKD. Here we report a case of coronary artery dissection occurring in a 38-year-old woman with ADPKD and an unremarkable medical history. She was admitted to the intensive care unit with acute chest pain. Anterior myocardial infarction was diagnosed by ECG; the patient was treated with thrombolysis with tenecteplase and aspirin, heparin and carvedilol. Then she was referred to the cardiac catheterization laboratory. Coronary angiography revealed a long linear dissection in the middle left descending coronary artery (DCA) with TIMI 1 distal flow. Complete vessel recanalization was achieved by positioning 2 drug-eluting and overlapped stents in the middle DCA and a further proximal stent due to residual proximal dissection, resulting in a TIMI 3 distal flow. There were no procedural complications, and the patient was discharged on day 8. In conclusion, spontaneous coronary artery dissection is a rare cause of myocardial ischemia and infarction. Myocardial infarction in patients without any risk factors for coronary atherosclerosis or in young patients should prompt a diligent search for a possible spontaneous coronary artery dissection. It is suggested that ADPKD may be a predisposing factor for spontaneous coronary artery dissection. PMID- 19557718 TI - BMP-7 counteracts TGF-beta1-induced epithelial-to-mesenchymal transition in human renal proximal tubular epithelial cells. AB - BACKGROUND: A large proportion of interstitial fibroblasts actually originate from tubular epithelial cells via the epithelial-to-mesenchymal transition (EMT) in renal fibrogenesis. Transforming growth factor-beta1 (TGF-beta1) is capable of initiating and completing the entire EMT course. Bone morphogenetic protein-7 (BMP-7) is a member of the TGF-beta superfamily. Recent studies indicate that BMP 7 could reverse established renal fibrosis in mice, primarily through counteracting TGF-beta1-mediated EMT. Therefore, we tested the hypothesis that BMP-7 functions by antagonizing profibrogenic events that are induced by TGF beta1 in cultured human renal proximal tubular epithelial (HK-2) cells. METHODS: Cultured HK-2 cells were treated with TGF-beta1 (3 ng/mL) or a combination of TGF beta1 and BMP-7 (100-400 ng/mL) for 48 hours. Morphological changes were assessed by phase contrast microscopy. The expression of alpha-smooth muscle actin (alpha SMA), E-cadherin, fibronectin, collagen I and connective tissue growth factor (CTGF) was analyzed by immunofluorescence, reverse transcriptase polymerase chain reaction and Western blotting. RESULTS: Incubation of HK-2 cells with 3 ng/mL TGF beta1 for 48 hours induced EMT, in association with decreased E-cadherin expression, increased alpha-SMA, fibronectin, collagen I and CTGF expression, and loss of epithelial morphology. BMP-7 inhibited all these effects in a dose dependent manner. In addition, 200 ng/mL BMP-7 reversed TGF-beta1-induced EMT, in association with reexpression of endogenous E-cadherin. CONCLUSIONS: These data suggest that BMP-7 attenuates progressive loss of kidney function and renal fibrosis through counteracting TGF-beta1-mediated EMT. PMID- 19557721 TI - Myeloperoxidase antineutrophil cytoplasmic antibody-associated vasculitis with diffuse tubulointerstitial nephritis. AB - Renal involvement in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis is characterized by focal segmental crescentic and/or necrotizing glomerulonephritis. Here, we report the case of a 66-year-old woman showing myeloperoxidase (MPO)-ANCA positivity and mononeuritis multiplex whose kidney biopsy revealed severe and diffuse tubulointerstitial nephritis despite the fact that crescentic necrotizing glomerulonephritis was focal. The mechanism of tubulointerstitial injury in ANCA-associated vasculitis remains unclear. Further studies are necessary to confirm the relationship between diffuse tubulointerstitial nephritis and ANCA-associated vasculitis. PMID- 19557723 TI - Mutants of the pentose-fermenting yeast Pichia stipitis with improved tolerance to inhibitors in hardwood spent sulfite liquor. AB - Mutants of Pichia stipitis NRRL Y-7124 able to tolerate and produce ethanol from hardwood spent sulfite liquor (HW SSL) were obtained by UV mutagenesis. P. stipitis cells were subjected to three successive rounds of UV mutagenesis, each followed by screening first on HW SSL gradient plates and then in diluted liquid HW SSL. Six third generation mutants with greater tolerance to HW SSL as compared to the wild type (WT) were isolated. The WT strain could not grow in HW SSL unless it was diluted to 65% (v/v). In contrast, the third generation mutants were able to grow in HW SSL diluted to 75% (v/v). Mutants PS301 and PS302 survived even in 80% (v/v) HW SSL, although there was no increase in cell number. All the third generation mutants exhibited higher growth rates but significantly lower growth yields on xylose or glucose compared to the WT. The mutants fermented 4% (w/v) glucose as efficiently as the WT and fermented 4% (w/v) xylose more efficiently with a higher ethanol yield than the WT. In a medium containing 4% (w/v) each of xylose and glucose, all the third generation mutants utilized glucose as efficiently and xylose more efficiently than the WT. This resulted in higher ethanol yield by the mutants. The mutants retained the ability to utilize galactose and mannose and ferment them to ethanol. Arabinose was consumed slowly by both the mutants and WT with no ethanol production. In 60% (v/v) HW SSL, the mutants utilized and fermented glucose, mannose, galactose and xylose while the WT could not ferment any of these sugars. PMID- 19557724 TI - Weed control in glyphosate-tolerant maize in Europe. AB - Maize growing in the EU27 increased to over 13 million ha in 2007, most of which (>80%) was grown in just eight countries (France, Romania, Germany, Hungary, Italy, Poland, Spain and Bulgaria). The number of herbicides used to control the wide spectrum of weeds occurring in all these countries is likely to decline in the future as each current active ingredient is reassessed for toxicological and environmental safety under Directive 91/414/EEC. Glyphosate has already been approved under this directive. Glyphosate, applied alone or in combination with currently available residual herbicides to genetically modified varieties tolerant to glyphosate, can provide a viable, flexible and profitable alternative to conventional weed control programmes. Glyphosate usage with glyphosate tolerant varieties also provides an environmentally sustainable weed control option as long as sufficient diversity of weed management options (crop rotation, chemical diversity, multiple cultural and mechanical practices, buffer strips) is maintained within the farm management system. Appropriate product stewardship measures will be required to maximise the long-term overall benefits of the glyphosate-based system. Specifically, care will need to be taken to manage potential weed shifts to more difficult-to-control species and to reduce the risk of selection for glyphosate-resistant weeds. PMID- 19557725 TI - Proparacaine complexation with beta-cyclodextrin and p-sulfonic acid calix[6]arene, as evaluated by varied (1)H-NMR approaches. AB - This study focused on the use of NMR techniques as a tool for the investigation of complex formation between proparacaine and cyclodextrins (CDs) or p-sulfonic acid calix[6]arene. The pH dependence of the complexation of proparacaine with beta-CD and p-sulfonic acid calix[6]arene was studied and binding constants were determined by (1)H NMR spectroscopy [diffusion-ordered spectroscopy (DOSY)] for the charged and uncharged forms of the local anesthetic in beta-CD and p-sulfonic acid calix[6]arene. The stoichiometries of the complexes was determined and rotating frame Overhauser enhancement spectroscopy (ROESY) 1D experiments revealed details of the molecular insertion of proparacaine into the beta-CD and p-sulfonic acid calix[6]arene cavities. The results unambiguously demonstrate that pH is an important factor for the development of supramolecular architectures based on beta-CD and p-sulfonic acid calix[6]arene as the host molecules. Such host-guest complexes were investigated in view of their potential use as new therapeutic formulations, designed to increase the bioavailability and/or to decrease the systemic toxicity of proparacaine in anesthesia procedures. PMID- 19557726 TI - MR prediction of postnatal outcomes in left-sided congenital diaphragmatic hernia using right lung signal intensity: comparison with that using right lung volume. AB - PURPOSE: To investigate the validity of the fetal right lung-to-liver signal intensity ratio (LLSIR) for prediction of postnatal outcomes in left-sided congenital diaphragmatic hernia (CDH) MATERIALS AND METHODS: The study included 14 pregnant women who underwent MR exams for evaluation of fetal left-sided CDH. The fetuses were divided into two groups: Group A (n = 9), alive, and Group B (n = 5), dead. On the basis of the half-Fourier acquisition single-shot turbo spin echo sequence, LLSIR and the right fetal lung volume (FLV) was calculated. In the control group, a regression analysis was performed to associate LLSIR and right FLV with gestational age. The relative LLSIR and right FLV (the observed/expected LLSIR and right FLV) were compared between Groups A and B. RESULTS: The mean relative LLSIR, as well as the mean relative right FLV, of Group A was significantly higher than that of Group B (p = 0.035). Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for the relative LLSIR and the relative right FLV, when the cutoff point was defined as 0.646 and 0.420, were the same and were 88.9%, 80.0%, 88.9%, 80.0%, and 85.7%, respectively. CONCLUSION: The postnatal outcomes in left-sided CDH may be predicted using the LLSIR. PMID- 19557727 TI - Temporal sampling requirements for reference region modeling of DCE-MRI data in human breast cancer. AB - PURPOSE: To assess the temporal sampling requirements needed for quantitative analysis of dynamic contrast-enhanced MRI (DCE-MRI) data with a reference region (RR) model in human breast cancer. MATERIALS AND METHODS: Simulations were used to study errors in pharmacokinetic parameters (K(trans) and v(e)) estimated by the RR model using six DCE-MRI acquisitions over a range of pharmacokinetic parameter values, arterial input functions, and temporal samplings. DCE-MRI data were acquired on 12 breast cancer patients and parameters were estimated using the native resolution data (16.4 seconds) and compared to downsampled 32.8-second and 65.6-second data. RESULTS: Simulations show that, in the majority of parameter combinations, the RR model results in an error less than 20% in the extracted parameters with temporal sampling as poor as 35.6 seconds. The experimental results show a high correlation between K(trans) and v(e) estimates from data acquired at 16.4-second temporal resolution compared to the downsampled 32.8-second data: the slope of the regression line was 1.025 (95% confidence interval [CI]: 1.021, 1.029), Pearson's correlation r = 0.943 (95% CI: 0.940, 0.945) for K(trans), and 1.023 (95% CI: 1.021. 1.025), r = 0.979 (95% CI: 0.978, 0.980) for v(e). For the 64-second temporal resolution data the results were: 0.890 (95% CI: 0.894, 0.905), r = 0.8645, (95% CI: 0.858, 0.871) for K(trans), and 1.041 (95% CI: 1.039, 1.043), r = 0.970 (95% CI: 0.968, 0.971) for v(e). CONCLUSION: RR analysis allows for a significant reduction in temporal sampling requirements and this lends itself to analyze DCE-MRI data acquired in practical situations. PMID- 19557728 TI - Pilot study of improved lesion characterization in breast MRI using a 3D radial balanced SSFP technique with isotropic resolution and efficient fat-water separation. AB - PURPOSE: To assess a 3D radial balanced steady-state free precession (SSFP) technique that provides submillimeter isotropic resolution and inherently registered fat and water image volumes in comparison to conventional T2-weighted RARE imaging for lesion characterization in breast magnetic resonance imaging (MRI). MATERIALS AND METHODS: 3D projection SSFP (3DPR-SSFP) combines a dual half echo radial k-space trajectory with a linear combination fat/water separation technique (linear combination SSFP). A pilot study was performed in 20 patients to assess fat suppression and depiction of lesion morphology using 3DPR-SSFP. For all patients fat suppression was measured for the 3DPR-SSFP image volumes and depiction of lesion morphology was compared against corresponding T2-weighted fast spin echo (FSE) datasets for 15 lesions in 11 patients. RESULTS: The isotropic 0.63 mm resolution of the 3DPR-SSFP sequence demonstrated improved depiction of lesion morphology in comparison to FSE. The 3DPR-SSFP fat and water datasets were available in a 5-minute scan time while average fat suppression with 3DPR-SSFP was 71% across all 20 patients. CONCLUSION: 3DPR-SSFP has the potential to improve the lesion characterization information available in breast MRI, particularly in comparison to conventional FSE. A larger study is warranted to quantify the effect of 3DPR-SSFP on specificity. PMID- 19557730 TI - Uses of proton pump inhibitors and serum potassium levels. AB - PURPOSE: Proton pump inhibitor (PPI) may suppress adrenal cortical steroid synthesis and release, thereby leading to electrolyte disturbances. Both hyponatremia and hyperkalemia in the setting of PPI therapy have been documented in case reports. The objective of this study was to examine the association between serum potassium (K(+)) level and PPI use. METHODS: A retrospective data analysis of hospitalized adults aged > or = 65 years during 2006, including PPI users (N = 257) and PPI non-users (N = 388), was conducted. Multiple linear and logistic regression analyses were used to assess the association between PPI use and serum K(+) level. RESULTS: PPI users [mean age (SD):79.7 (8.0) years; 70% female] had significantly higher serum K(+) levels than PPI non-users [80.2 (8.8) years; 64% female] on admission [4.13 (0.62) vs. 3.97 (0.57) mmol/L; p < 0.001]. The linear regression model revealed that > or = 2 defined daily dose (DDD) units of PPI use were a significantly positive contributor to serum K(+) levels (p = 0.021) after adjusting for age, serum creatinine levels, sex, history of diabetes, and uses of the following drugs: angiotensin-converting enzyme (ACE) inhibitor, angiotensin receptor blocker, beta blocker, diuretics, spironolactone, K(+) supplement, non-steroidal anti-inflammatory drugs, atypical antipsychotics, and narcotics. However, multiple logistic regression model revealed that high dose PPI therapy was not associated with an increased risk for hyperkalemia occurrence (p = 0.762). CONCLUSION: Higher serum K(+) levels were observed among PPI users when compared to PPI non-users. High daily dose PPI therapy may be an independent positive predictor of serum potassium levels. PMID- 19557729 TI - Bio-electrospraying whole human blood: analysing cellular viability at a molecular level. AB - Bio-electrosprays, pioneered in 2005, have undergone several developmental studies which have seen this technique evolve as a novel direct in vivo tissue engineering and regenerative medicinal strategy. Those studies have been a hallmark for electrosprays; however, in this communication we report our on-going developmental investigations for exploring bio-electrosprays as a potential medical device and diagnostic protocol. The studies reported here demonstrate the ability to directly jet whole human blood without affecting the genetic make-up, which has been interrogated by way of reverse transcription-polymerase chain reaction (RT-PCR) in comparison to controls (p = 0.7337). These studies demonstrate bio-electrosprays as a possible diagnostic protocol. PMID- 19557731 TI - Transition metal NMR chemical shifts and polarizability effect in organometallic complexes. AB - The literature data on substituent influence on the (51)V, (55)Mn, (57)Fe, (59)Co, (61)Ni, (95)Mo, (103)Rh, (183)W, (187)Os and (195)Pt NMR chemical shifts (delta) and on J (M, P; M = Mn, Fe, Mo, Rh, W, Os) coupling constants have been analyzed for 30 series of the organometallic complexes. It has been established for the first time that the delta and J values depend on the inductive, resonance and polarizability effects of substituents. The polarizability effect is caused by the partial charge on the central M atom. The contribution of this effect ranges from 3 to 86%. PMID- 19557732 TI - Computerized characterization of prostate cancer by fractal analysis in MR images. AB - PURPOSE: To explore the potential of computerized characterization of prostate MR images by extracting the fractal features of texture and intensity distributions as indices in the differential diagnosis of prostate cancer. MATERIALS AND METHODS: MR T2-weighted images (T2WI) of 55 patients with pathologic results detected by ultrasound guided biopsy were collected and then divided in two groups, 27 with prostate cancer (PCa) and 28 with no histological abnormality. Texture fractal dimension (TFD) and histogram fractal dimension (HFD) were calculated to analyze complexity features of regions of Interest (ROIs) selected from the peripheral zone. Two-sample t-tests were performed to evaluate group differences for both parameters. Receiver operating characteristic (ROC) analysis was used to estimate the performance of TFD and HFD for discriminating PCa. RESULTS: Significant differences were found in both TFD and HFD between the two patient groups. The areas under the ROC curves of TFD and HFD were 0.691 and 0.966, respectively, in distinguishing prostatic carcinoma from normal peripheral zone. As characterized by the fractal indices, cancerous prostatic tissue exhibited smoother texture and lower variation in intensity distribution than normal prostatic tissue. CONCLUSION: The study suggests that TFD and HFD depict the changes in texture and intensity distribution associated with prostate cancer on T2WI. Both TFD and HFD provide promising quantitative indices for cancer identification. HFD performs better than TFD offering a more robust MR-based indicator in the diagnosis of prostatic carcinoma. PMID- 19557733 TI - Effect of PRESS and STEAM sequences on magnetic resonance spectroscopic liver fat quantification. AB - PURPOSE: To compare PRESS and STEAM MR spectroscopy for assessment of liver fat in human subjects. MATERIALS AND METHODS: Single-voxel (20 x 20 x 20 mm) PRESS and STEAM spectra were obtained at 1.5T in 49 human subjects with known or suspected fatty liver disease. PRESS and STEAM sequences were obtained with fixed TR (1500 msec) and different TE (five PRESS spectra between TE 30-70 msec, five STEAM spectra between TE 20-60 msec). Spectra were quantified and T2 and T2 corrected peak area were calculated by different techniques. The values were compared for PRESS and STEAM. RESULTS: Water T2 values from PRESS and STEAM were not significantly different (P = 0.33). Fat peak T2s were 25%-50% shorter on PRESS than on STEAM (P < 0.02 for all comparisons) and there was no correlation between T2s of individual peaks. PRESS systematically overestimated the relative fat peak areas (by 7%-263%) compared to STEAM (P < 0.005 for all comparisons). The peak area given by PRESS was more dependent on the T2-correction technique than STEAM. CONCLUSION: Measured liver fat depends on the MRS sequence used. Compared to STEAM, PRESS underestimates T2 values of fat, overestimates fat fraction, and provides a less consistent fat fraction estimate, probably due to J coupling effects. PMID- 19557734 TI - Added value of diffusion-weighted imaging in the MRI assessment of perilesional tumor recurrence after chemoembolization of hepatocellular carcinomas. AB - PURPOSE: To investigate the additional value of diffusion-weighted imaging (DWI) in the MRI assessment of perilesionally recurrent hepatocellular carcinomas (HCCs) after transcatheter arterial chemoembolization (TACE). MATERIALS AND METHODS: For gadopentetate dimeglumine-enhanced multiphasic dynamic images with precontrast T1- and T2-weighted images of 23 patients treated with TACE, two radiologists independently sorted the confidence levels for the presence of a viable tumor in the vicinity of the treated lesions into five grades. In another session, images from DWI (b factor = 50, 400, and 800 s/mm(2)) were added to the previously reviewed images and the same radiologists sorted the confidence levels. RESULTS: A total of 26 lesions (0.7-3.5 cm) from 16 patients were confirmed to be perilesional recurrences of HCC. The area under the receiver operating characteristic curve (A(z)) for the second interpretation session (0.826) was not significantly different (P = 0.299) from that of the first session (0.859). The overall sensitivity was increased from 85% to 92%, but the specificity decreased from 65% to 50% after adding DWI. CONCLUSION: The addition of DWI has the potential to improve sensitivity, but not the overall diagnostic accuracy, in the assessment of perilesional recurrence of HCCs after chemoembolization. PMID- 19557735 TI - Gene expression profiling during early response to injury and microbial challenges in the silkworm, Bombyx mori. AB - To identify Bombyx mori genes involved in the early response to injury and microbial challenge, we performed genome-wide gene expression-profiling experiments using oligonucleotide DNA microarrays. Of approximately 23,000 genes examined, 465 displayed changes in mRNA expression levels. Of these, 306 were induced and 159 were repressed in response to injury (injection with phosphate buffer saline) or challenges by Gram-negative (Serratia marcescens), Gram positive bacteria (Staphylococcus aureus), or fungus (Beauveria bassiana). Many of these differentially expressed genes can be assigned to specific functional groups of the innate immune response, including recognition, signaling, melanization and coagulation, and antimicrobial peptides. Seventeen percent of differentially expressed genes encode proteins with no obvious similarity to known functional domains. Of particular interest is a member of the juvenile hormone-binding protein family, which was highly induced by both injury and microbial challenges. The possible role of juvenile hormone in innate immunity is discussed. PMID- 19557736 TI - Failure of beta-cell function to compensate lack of insulin action in hyperuricemic subjects. AB - BACKGROUND: Elevated serum uric acid has been related to insulin resistance, but there are no studies regarding its association with beta-cell function. In this study, we examine the response of beta-cell function to compensate insulin resistance in hyperuricaemic subjects. METHODS: Population-based cross-sectional study. A total of 212 healthy individuals, men and non-pregnant women, aged 20-65 years, were randomly recruited and allocated into groups with hyperuricaemia or normouricaemia that were matched by value of Belfiore index. Hyperuricaemia was defined by serum uric acid concentration >or=416 micromol/L in men and >or=357 micromol/L in women. To evaluate the compensation of insulin secretion to variation of insulin sensitivity, we used a surrogate of the hyperbolic model of beta-cell function based on Belfiore and Homeostasis Model Analysis beta-cell (HOMA-ss) indexes; the area under curves (AUC) was calculated. The association between the uric acid levels and the AUC was estimated using a multivariate regression analysis adjusted by age, sex, and waist circumference. RESULTS: Hyperuricaemia was identified in 77 (36.3%) individuals. A total of 39 (50.6%) hyperuricaemic and 44 (32.6%) normouricaemic subjects had diagnosis of impaired glucose tolerance (IGT), p = 0.01. In the total population, the AUCs in the hyperuricaemic and normouricaemic groups were 1.60725 cm(2) and 3.9819 cm(2), respectively (odds ratio [OR] 3.9; CI 95% 1.4-8.9). Among the 83 individuals with IGT, the AUCs in the subgroups with and without hyperuricaemia were 0.8495 cm(2) and 2.42075 cm(2), respectively (OR 2.7; CI 95% 1.3-10.1). CONCLUSIONS: Our results show the failure of beta-cell function to compensate variation of insulin sensitivity in the hyperuricaemic individuals. PMID- 19557737 TI - SAR and power implications of different RF shimming strategies in the pelvis for 7T MRI. AB - PURPOSE: To determine the best radiofrequency (RF) shimming method for 7 T body imaging that provides sufficient B(1)(+) excitation inside the target region while energy deposition (SAR) and power demands are as low as possible and that does not incorporate anatomy specific electric field information inside the patient models, as this information is not available in practice. MATERIALS AND METHODS: Finite difference time domain (FDTD) simulations were used to evaluate five RF shimming strategies for the pelvis inside a body coil. The results were compared to the theoretical best solution that could be achieved if the electric field inside the patient was known. RESULTS: Most of the RF shimming strategies were successful. However, between the different strategies a factor of two difference in average SAR reduction, a factor of three difference in local maximum SAR reduction, and a factor of 20 difference in power efficiency was observed. Phase matching was found to be the most promising RF shimming method for the body coil used and patient models. CONCLUSION: RF shimming can reduce the SAR and improve power efficiency in an accurate patient model without knowing the electric field. However, choosing the right method is critical to prevent unexpected behavior in local SAR deposition. PMID- 19557738 TI - Improved myelin water quantification using spatially regularized non-negative least squares algorithm. AB - PURPOSE: To improve the myelin water quantification in the brain in the presence of measurement noise and to increase the visibility of small focal lesions in myelin-water-fraction (MWF) maps. MATERIALS AND METHODS: A spatially regularized non-negative least squares (srNNLS) algorithm was developed for robust myelin water quantification in the brain. The regularization for the conventional NNLS algorithm was expanded into the spatial domain in addition to the spectral domain. Synthetic data simulations were performed to study the effectiveness of this new algorithm. Experimental free-induction-decay measurements were obtained using a multi-gradient-echo pulse sequence and MWF maps were estimated using the srNNLS algorithm. The results were compared with other conventional methods. RESULTS: A substantial decrease in MWF variability was observed in both simulations and experimental data when the srNNLS algorithm was applied. As a result, false lesions in the MWF maps disappeared and the visibility of small focal lesions improved greatly. On average, the contrast-to-noise ratio for focal lesions was improved by a factor of 2. CONCLUSION: The MWF variability due to the measurement noise can be substantially reduced and the detection of small focal lesions can be improved by using the srNNLS algorithm. PMID- 19557739 TI - Esterase-based resistance in the tobacco-adapted form of the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae) in the eastern United States. AB - Organophosphates and carbamates represent alternative insecticides in managing the tobacco-adapted form of the green peach aphid (TGPA), Myzus persicae (Sulzer), a major pest of tobacco in the United States and around the world. General esterases that detoxify these insecticides were assessed in green, red, and orange morphs of field-collected M. persicae. A total of 136 aphid colonies were collected from 2004 though 2007 and screened for total esterase activity. The green morphs had lower esterase levels, with a mean of 77+/-6.6 nmol/min/mg protein, as compared to red (84+/-2.9 nmol/min/mg protein) and orange morphs (172+/-16.5 nmol/min/mg protein). Overall esterase activities, and those for the red and green morphs, were positively correlated with LC(50) values for acephate (organophosphate) and methomyl (carbamate) assessed in leaf-dip bioassays. Esterase genes responsible for higher esterase activities were diagnosed by gene amplification studies. Twenty-three of 24 colonies tested had either the E4 or FE4 gene amplified, both known to confer esterase-based resistance. Fifteen out of the 24 colonies tested had amplified E4 gene and four colonies had FE4 gene amplification. All orange morphs and one green morph had both E4 and FE4 genes amplified. This unique phenotype, where two esterase genes were amplified had an 865-bp band characteristic of the FE4 gene and an additional 381-bp band characteristic of a deleted upstream region of the E4 gene. Changes that occurred in esterase-based resistance in the TGPA over the past two decades and their implications on insecticide resistance management are discussed. PMID- 19557740 TI - Automated assessment of whole-body adipose tissue depots from continuously moving bed MRI: a feasibility study. AB - PURPOSE: To present an automated algorithm for segmentation of visceral, subcutaneous, and total volumes of adipose tissue depots (VAT, SAT, TAT) from whole-body MRI data sets and to investigate the VAT segmentation accuracy and the reproducibility of all depot assessments. MATERIALS AND METHODS: Repeated measurements were performed on 24 volunteer subjects using a 1.5 Tesla clinical MRI scanner and a three-dimensional (3D) multi-gradient-echo sequence (resolution: 2.1 x 2.1 x 8 mm(3), acquisition time: 5 min 15 s). Fat and water images were reconstructed, and fully automated segmentation was performed. Manual segmentation of the VAT reference was performed by an experienced operator. RESULTS: Strong correlation (R = 0.999) was found between the automated and manual VAT assessments. The automated results underestimated VAT with 4.7 +/- 4.4%. The accuracy was 88 +/- 4.5% and 7.6 +/- 5.7% for true positive and false positive fractions, respectively. Coefficients of variation from the repeated measurements were: 2.32 % +/- 2.61%, 2.25% +/- 2.10%, and 1.01% +/- 0.74% for VAT, SAT, and TAT, respectively. CONCLUSION: Automated and manual VAT results correlated strongly. The assessments of all depots were highly reproducible. The acquisition and postprocessing techniques presented are likely useful in obesity related studies. PMID- 19557741 TI - Characterization of low-grade gliomas using RGB color maps derived from ADC histograms. AB - PURPOSE: To use normalized apparent diffusion coefficient (nADC) histograms from patients with grade II oligodendroglioma (OD) and astrocytoma (AC) to generate RGB color maps that emphasize the differences between normal-appearing white matter (NAWM), oligo-like, and astro-like regions. MATERIALS AND METHODS: NAWM and nonenhancing lesion (NEL) ADC values from 19 ODs and 11 ACs were summed to generate oligo-like (red), NAWM (green), and astro-like (blue) nADC histograms. These nADC histograms were then used to map nADC values to an RGB matrix. RESULTS: Color maps of oligodendroglial tumor regions were generally visualized in pink, while color maps of astrocytic tumor regions showed various shades of blue. This technique was also applied to 23 patients with the more mixed subtype, oligoastrocytoma (OA), which showed a mixture of both blue and pink, which in many cases appeared to bleed into each other and were blotchy. CONCLUSION: This technique allows for the visualization of biologically different regions within the whole tumor mass, which may aid in directing image-guided biopsies. This can be used to ensure that the biopsy is directed to regions that can more accurately define the dominant tumor characteristics. PMID- 19557742 TI - Sodium time course using 23Na MRI in reversible focal brain ischemia in the monkey. AB - PURPOSE: To demonstrate the use of sodium MRI for measuring the time course of tissue sodium concentration (TSC) in a nonhuman primate model of reversible focal brain ischemia. MATERIALS AND METHODS: Reversible endovascular focal brain ischemia was induced in nonhuman primates (n = 4), and sodium MRI was performed on a 3 Tesla scanner for monitoring changes in TSC during both the middle cerebral artery (MCA) occlusion and MCA reperfusion portions of the experiment. RESULTS: The TSC increased linearly in the ischemic tissue during MCA occlusion (ranging from a mean TSC increase of 5.44%/h to 7.15%/h across the four subjects), and then there was a statistically significant change from a positive TSC slope during MCA occlusion to a TSC slope after MCA reperfusion that was not statistically different from zero. The linear increase in sodium MRI during brain ischemia was used to estimate the stroke onset time to within 0.45 h in each of the four subjects (with a maximum 95% confidence interval of +/- 1.147 h). CONCLUSION: The data indicate that sodium MRI increases linearly during brain ischemia, and that this increase is stopped by tissue reperfusion within 5.4 h after stroke onset. PMID- 19557744 TI - Reproducibility of dGEMRIC in assessment of hip joint cartilage: a prospective study. AB - PURPOSE: To investigate the reproducibility of dGEMRIC in the assessment of cartilage health of the adult asymptomatic hip joint. MATERIALS AND METHODS: Fifteen asymptomatic volunteers (mean age, 26.3 years +/- 3.0) were preliminarily studied. Any volunteer that was incidentally diagnosed with damaged cartilage on MRI (n = 5) was excluded. Ten patients that had no evidence of prior cartilage damage (mean age, 26.2 years +/- 3.4) were evaluated further in this study. The reproducibility of dGEMRIC was assessed with two T1(Gd) exams performed 4 weeks apart in these volunteers. The protocol involved an initial standard MRI to confirm healthy cartilage, which was then followed by dGEMRIC. The second scan included only the repeat dGEMRIC. Region of interest (ROI) analyses for T1(Gd) measurement was performed in seven radial reformats. Statistical analysis included the student's t-test and intra-class correlation (ICC) measurement to assess reproducibility. RESULTS: Overall 70 ROIs were studied. Mean cartilage T1(Gd) values at various loci ranged from 560.9 ms to 684.4 ms at the first set of readings and 551.5 ms to 662.2 ms in the second one. The mean difference per region of interest between the two T1(Gd)-measurements ranged from 21.4 ms (3.7%) to 45.0 ms (6.8%), which was not found to be statistically significant (P = 0.153). There was a high reproducibility detected (ICC range, 0.667-0.915). Intra and Inter-observer analyses proved a high agreement for T1(Gd) assessment (0.973 and 0.932). CONCLUSION: We found dGEMRIC to be a reliable tool in the assessment of cartilage health status in adult hip joints. PMID- 19557743 TI - High b-value apparent diffusion-weighted images from CURVE-ball DTI. AB - PURPOSE: To investigate the utility of a proposed clinical diffusion imaging scheme for rapidly generating multiple b-value diffusion contrast in brain magnetic resonance imaging (MRI) with high signal-to-noise ratio (SNR). MATERIALS AND METHODS: Our strategy for efficient image acquisition relies on the invariance property of the diffusion tensor eigenvectors to b-value. A simple addition to the conventional diffusion tensor MR imaging (DTI) data acquisition scheme used for tractography yields diffusion-weighted images at twice and three times the conventional b-value. An example from a neurosurgical brain tumor is shown. Apparent diffusion-weighted (ADW) images were calculated for b-values 800, 1600, and 2400 s/mm(2), and a map of excess diffusive kurtosis was computed from the three ADWs. RESULTS: High b-value ADW images demonstrated decreased contrast between normal gray and white matter, while the heterogeneity and contrast of the lesion was emphasized relative to conventional b-value data. Kurtosis maps indicated the deviation from Gaussian diffusive behavior. CONCLUSION: DTI data with multiple b-values and good SNR can be acquired in clinically reasonable times. High b-value ADW images show increased contrast and add information to conventional DWI. Ambiguity in conventional b-value images over whether hyperintense signal results from abnormally low diffusion, or abnormally long T(2), is better resolved in high b-value images. PMID- 19557745 TI - Maximized PUFA measurements improve insight in changes in fatty acid composition in response to temperature. AB - A general mechanism underlying the response of ectotherms to environmental changes often involves changes in fatty acid composition. Theory predicts that a decrease in temperature causes an increase in unsaturation of fatty acids, with an important role for long-chain poly-unsaturated fatty acids (PUFAs). However, PUFAs are particularly unstable and susceptible to peroxidation, hence subtle differences in fatty acid composition can be challenging to detect. We determined the fatty acid composition in springtail (Collembola) in response to two temperatures (5 degrees C and 25 degrees C). First, we tested different sample preparation methods to maximize PUFAs. Treatments consisted of different solvents for primary lipid extraction, mixing with antioxidant, flushing with inert gas, and using different temperature exposures during saponification. Especially slow saponification at low temperature (90 min at 70 degrees C) in combination with replacement of headspace air with nitrogen during saponification and methylation maximized PUFAs for GC analysis. Applying these methods to measure thermal responses in fatty acid composition, the data showed that the (maximized) proportion of C(20) PUFAs increased at low acclimation temperature. However, C(18) PUFAs increased at high acclimation temperature, which is contrary to expectations. Our study illustrates that PUFA levels in lipids may often be underestimated and this may hamper a correct interpretation of differential responses of fatty acid composition. PMID- 19557746 TI - Automated estimation of regional mean transition times and radial velocities from cine magnetic resonance images: evaluation in normal subjects. AB - PURPOSE: To assess regional ventricular function via an accurate and automated definition of functional parameters. MATERIALS AND METHODS: An automated method is proposed that estimates reliable regional normalized mean transition times (F(mc)) and mean radial velocities (V(m)) from cine images. This approach combines a quantitative parametric imaging method and an automated detection of the endocardial border, which is robust to the presence of papillary muscles and nonhomogeneities within the left ventricular cavity. Steady-state free-precession cine-magnetic resonance imaging (MRI) of 36 healthy subjects was analyzed. RESULTS: The quality of the automated segmentation was assessed on a subgroup of 20 subjects by comparing the results with the manual contours traced by an expert. The comparison of functional parameters estimated consequently using the automated and the manual contours yielded (y = 0.959x + 0.016, R = 0.964) for F(mc) and (y = 0.883x + 0.505, R = 0.935) for V(m). On the entire group, F(mc) was equal to 0.392 +/- 0.069 and V(m) to 5.4 +/- 2.3 cm/s. Increasing values of the temporal parameter from the apex to the base and larger values in the septal wall than in lateral wall were demonstrated and were in accordance with the physiology. CONCLUSION: The proposed method ensures an automated and robust assessment of regional wall motion parameters, which could be clinically useful. PMID- 19557747 TI - Characterization of the trypsin-like protease (Ha-TLP2) constitutively expressed in the integument of the cotton bollworm, Helicoverpa armigera. AB - Trypsins belong to the serine endoproteases. They are the most important proteases in insects because of their key roles in food digestion and zymogens activation. But there has been little study of the trypsins in the integuments of insects. In this work, we cloned a trypsin-like protease gene from Helicoverpa armigera and named it trypsin-like protease 2 (Ha-TLP2). Semi-quantitative reverse transcription PCR analysis showed that Ha-TLP2 is constitutively expressed in the integument and can be down-regulated by 20-hydroxyecdysone (20E) and up-regulated by the juvenile hormone (JH) analog methoprene. Immunohistochemistry showed that Ha-TLP2 is located not only in the epidermis, but also in new and old cuticles. Immunoblotting and gelatin-SDS-PAGE revealed that Ha-TLP2 is constitutively expressed with activity in the integument during larval feeding, molting, and metamorphosis. This evidence suggests that Ha-TLP2 is involved in the remodeling of the integument. PMID- 19557748 TI - Direct MR arthrography of the shoulder at 3 Tesla: optimization of gadolinium concentration. AB - PURPOSE: To determine the optimal concentration of gadopentetate dimeglumine for direct shoulder MR arthrography at 3T in vivo. MATERIALS AND METHODS: Sixty-eight consecutive shoulder MR arthrograms were obtained with 1 mmolGd/L (n = 12), 2 mmolGd/L (n = 12), 4 mmolGd/L (n = 12), 6 mmolGd/L (n = 12), 9 mmolGd/L (n = 10) and 12 mmolGd/L (n = 10). All postinjection fat-suppressed T1-weighted and T2 weighted images were analyzed retrospectively. For qualitative evaluation, image contrast was graded on a subjective three-level scale (excellent, moderate, and poor). For quantitative analysis, the contrast-to-noise ratio (CNR) of intra articular fluid to muscle was measured. RESULTS: All postinjection T1-weighted images and T2-weighted images with 1 mmolGd/L, 2 mmolGd/L, 4 mmolGd/L, and 6 mmolGd/L were qualitatively evaluated as excellent or moderate. Two of the ten 9 mmolGd/L images and seven of the ten 12-mmolGd/L images were rated as poor with regard to the T2 image contrast. On the T1-weighted images, no significant difference existed between the CNRs of the six concentrations, but a peak CNR was seen at the concentration of 6 mmolGd/L. On the T2-weighted images, CNRs at concentrations of 1 mmolGd/L, 2 mmolGd/L, 4 mmolGd/L, 6 mmolGd/L, and 9 mmolGd/L showed no statistical difference, but were all significantly higher than that with 12 mmolGd/L. CONCLUSION: The acceptable concentration of gadopentetate dimeglumine for shoulder MR arthrography at 3T was found to be in the range of 1 mmolGd/L to 6 mmolGd/L. 6 mmolGd/L may be the optimal concentration. PMID- 19557749 TI - Expression, purification, and characterization of pro-phenoloxidase-activating serine protease from Spodoptera litura. AB - One of the important trigger molecules for innate immunity is a serine protease that activates zymogen phenol oxidase (PPO). Central to wound healing response is the activation of phenol oxidase zymogen. Molecular characterization of phenol oxidase has been recently reported by us. Here, we report isolation, cloning, expression, and purification of prophenol oxidase activating enzyme 1 (slppae1) from polyphagous pest, Spodoptera litura. SLPPAE1 is induced within 6 h of physical injury. The structural features of the mature polypeptide are reminiscent of other lepidopteran PPAE in having a signal peptide, propeptide, and catalytically active polypeptide. The cDNA has been expressed in Sf21 cells using baculovirus expression vector. Fractionation of expressing Sf21 cells revealed its expression in the membranes. The recombinant protein was solubilized from membranes and purified by Ni-NTA affinity chromatography. The purified enzyme is catalytically active on chromogenic substrate, activates recombinantly expressed prophenol oxidase (PPO) of S. litura, and is sensitive to inhibition by aprotenin. N-terminal sequencing of processed phenol oxidase revealed 11 kDa propeptide instead of in-silico predicted 6 kDa polypeptide. PMID- 19557751 TI - Single-marker and two-marker association tests for unphased case-control genotype data, with a power comparison. AB - In case-control single nucleotide polymorphism (SNP) data, the allele frequency, Hardy Weinberg Disequilibrium, and linkage disequilibrium (LD) contrast tests are three distinct sources of information about genetic association. While all three tests are typically developed in a retrospective context, we show that prospective logistic regression models may be developed that correspond conceptually to the retrospective tests. This approach provides a flexible framework for conducting a systematic series of association analyses using unphased genotype data and any number of covariates. For a single stage study, two single-marker tests and four two-marker tests are discussed. The true association models are derived and they allow us to understand why a model with only a linear term will generally fit well for a SNP in weak LD with a causal SNP, whatever the disease model, but not for a SNP in high LD with a non-additive disease SNP. We investigate the power of the association tests using real LD parameters from chromosome 11 in the HapMap CEU population data. Among the single marker tests, the allelic test has on average the most power in the case of an additive disease, but for dominant, recessive, and heterozygote disadvantage diseases, the genotypic test has the most power. Among the four two-marker tests, the Allelic-LD contrast test, which incorporates linear terms for two markers and their interaction term, provides the most reliable power overall for the cases studied. Therefore, our result supports incorporating an interaction term as well as linear terms in multi-marker tests. PMID- 19557750 TI - Docetaxel, cisplatin, and fluorouracil induction chemotherapy followed by accelerated fractionation/concomitant boost radiation and concurrent cisplatin in patients with advanced squamous cell head and neck cancer: A Southwest Oncology Group phase II trial (S0216). AB - BACKGROUND: In an effort to optimize nonoperative therapy in patients with locoregionally advanced head and neck squamous cell cancer, the Southwest Oncology Group conducted a phase II trial combining 3-drug taxane-containing induction chemotherapy with accelerated fractionation/concomitant boost radiation and concomitant single-agent cisplatin. METHODS: Two induction courses using docetaxel (75 mg/m(2) on day 1), cisplatin (100 mg/m(2) on day 1), and fluorouracil (1000 mg/m(2)/day continuous intravenous infusion days 1-4) were given, with an interval of 21 days. Patients who were stable or responded to the chemotherapy received definitive accelerated fractionation/concomitant boost radiation with concurrent cisplatin (100 mg/m(2)) on days 1 and 22 of radiation. RESULTS: There were 74 eligible and evaluable patients enrolled between March 1, 2003, and August 15, 2004; 52 (70%) had stage IV disease. At least 1 grade 3-4 toxicity was experienced by 63 patients (85%) during induction. A total of 61 patients completed induction and began concurrent chemoradiotherapy; 50 (68%) completed all planned treatment. At least 1 grade 3-4 toxicity was noted in 53 of the 58 patients (91%) evaluated for toxicity from concurrent chemoradiotherapy. Two patients died during induction, and 2 during chemoradiation. With a median follow-up of 36 months (range, 14-50), the 2-year and 3-year overall survival estimates were 70% and 64%, with 2-year and 3-year progression-free survival estimates of 66% and 61%, respectively. CONCLUSIONS: Three-drug induction chemotherapy followed by accelerated fractionation/concomitant boost radiation and concurrent cisplatin is toxic but feasible within a cooperative group. In this patient cohort with advanced head and neck squamous cell cancer, overall and progression-free survivals were encouraging, justifying further study of this approach. PMID- 19557752 TI - OCT imaging of skin cancer and other dermatological diseases. AB - Optical coherence tomography (OCT) provides clinicians and researchers with micrometer-resolution, in vivo, cross-sectional images of human skin up to several millimeter depth. This review of OCT imaging applied within dermatology covers the application of OCT to normal skin, and reports on a large number of applications in the fields of non-melanoma skin cancer, malignant melanomas, psoriasis and dermatitis, infestations, bullous skin diseases, tattoos, nails, haemangiomas, and other skin diseases. PMID- 19557756 TI - Enantioselective artificial metalloenzymes based on a bovine pancreatic polypeptide scaffold. AB - Site creation: Enantioselective artificial metalloenzymes have been created by grafting a new active site onto bovine pancreatic polypeptide through the introduction of an amino acid capable of coordinating a copper(II) ion. This hybrid catalyst gave good enantioselectivities in the Diels-Alder and Michael addition reactions in water (see scheme) and displayed a very high substrate selectivity. PMID- 19557758 TI - Who succeeds in jail diversion programs for persons with mental illness? A multi site study. AB - This study examined how the characteristics of people with mental illness who are participants in post-booking jail diversion programs affect recidivism and time spent incarcerated. The study employed data from a multi-site, federally funded jail diversion initiative. A pre-post comparison design was used to compare experiences of arrest and days spent in jail of diverted individuals for the 12 months following enrollment with the 12 months prior to enrollment. Also compared were differences in 12-month public safety outcomes. Data were collected between February 2003 and August 2007. The findings suggest that people with mental illness who are diverted from jail to community-based services experience fewer arrests and jail days. Approximately half of the sample were never arrested during the 12 months following enrollment. The strongest differences between people who experienced reduced contact with the criminal justice system and people with unchanged or increased contact were found in measures of criminal history. The results suggest that services targeted to diverted individuals with mental illness should address public safety goals, not just those of public health. PMID- 19557759 TI - Allosteric ligands for G protein-coupled receptors: a novel strategy with attractive therapeutic opportunities. AB - Allosteric receptor ligands bind to a recognition site that is distinct from the binding site of the endogenous messenger molecule. As a consequence, allosteric agents may attach to receptors that are already transmitter-bound. Ternary complex formation opens an avenue to qualitatively new drug actions at G protein coupled receptors (GPCRs), in particular receptor subtype selective potentiation of endogenous transmitter action. Consequently, suitable exploitation of allosteric recognition sites as alternative molecular targets could pave the way to a drug discovery paradigm different from those aimed at mimicking or blocking the effects of endogenous (orthosteric) receptor activators. The number of allosteric ligands reported to modulate GPCR function is steadily increasing and some have already reached routine clinical use. This review aims at introducing into this fascinating field of drug discovery and at providing an overview about the achievements that have already been made. Various case examples will be discussed in the framework of GPCR classification (family A, B, and C receptors). In addition, the behavior at muscarinic receptors of hybrid derivatives incorporating both an allosteric and an orthosteric fragment in a common molecular skeleton will be illustrated. PMID- 19557755 TI - Palladium(II)-catalyzed C-H activation/C-C cross-coupling reactions: versatility and practicality. AB - In the past decade, palladium-catalyzed C-H activation/C-C bond-forming reactions have emerged as promising new catalytic transformations; however, development in this field is still at an early stage compared to the state of the art in cross coupling reactions using aryl and alkyl halides. This Review begins with a brief introduction of four extensively investigated modes of catalysis for forming C-C bonds from C-H bonds: Pd(II)/Pd(0), Pd(II)/Pd(IV), Pd(0)/Pd(II)/Pd(IV), and Pd(0)/Pd(II) catalysis. A more detailed discussion is then directed towards the recent development of palladium(II)-catalyzed coupling of C-H bonds with organometallic reagents through a Pd(II)/Pd(0) catalytic cycle. Despite the progress made to date, improving the versatility and practicality of this new reaction remains a tremendous challenge. PMID- 19557760 TI - Analysis of novel over- and under-sulfated glycosaminoglycan sequences by enzyme cleavage and multiple stage MS. AB - We report on a novel strategy for identification of specific sulfation motifs in chondroitin/dermatan sulfate (CS/DS) chain derived from decorin (Dcn), based on enzyme cleavage and multistage MS (MS(n)). Released CS/DS chains were digested with chondroitin B and in parallel with AC I lyases to obtain oligosaccharides of known hexuronic acid (HexA) epimerization. The depolymerized chains were separated by gel filtration, and collected di- and hexasaccharides were analyzed by ESI MS(n). MS(2) on bisulfated 4,5-Delta-HexAGalNAc revealed an additional sulfate ester group at 4,5-Delta-HexA. MS(2) data provided evidence upon GlcA sulfation in Dcn due to the fact that 4,5-Delta-HexA derived from GlcA after chondroitin AC I lyase treatment. Hexasaccharide screening in the MS(1) mode indicated direct correlation between the sulfate distribution and HexA epimerization. MS(n) performed on ions that, according to mass calculation, correspond to pentasulfated [4,5-Delta-HexAGalNAc(GlcAGalNAc)(2)], trisulfated [4,5-Delta-HexAGalNAc(GlcAGalNAc)(2)] with IdoA-derived 4,5-Delta-HexA at the nonreducing end, tetrasulfated [4,5-Delta-HexAGalNAc(IdoAGalNAc)(2)] and monosulfated [4,5-Delta-HexAGalNAc(IdoAGalNAc)(2)] with GlcA-derived 4,5-Delta HexA at the nonreducing end rendered fragmentation patterns confirming the presence of over-, regular, and under-sulfated regions as well as structural motifs having both types of HexA sulfated within Dcn CS/DS. PMID- 19557761 TI - Comparative proteome profile of immature rat ovary during primordial follicle assembly and development. AB - The assembly of primordial follicles early in ovarian development and subsequent transition to primary follicles are critical processes in ovarian biology. Inappropriate coordination of these processes contributes to ovarian pathologies such as premature ovarian failure and infertility. To better understand the molecular mechanisms involved in primordial follicle assembly and development, 2 D PAGE and MALDI-TOF/TOF technologies were used to construct a comparative proteome profile of the immature rat ovary at specific time-points (0, 24, 48, and 72 h postpartum). A total of 154 differential protein spots corresponding to 134 different proteins were definitively identified between any two time-points. Further cluster analysis showed four expression patterns, and each pattern correlated with specific cell processes that occur during early ovarian development. Seven proteins were randomly selected to verify expression patterns using Western blotting, and subsequently immunohistochemistry was performed to further investigate their cellular localization. Additionally, detailed functional analyses of these differentially expressed proteins were performed. Elucidation of how these changes in protein expression level coordinate primordial follicles assembly and development is intended to provide a better understanding of these critical biological processes early in ovarian development and will provide potential therapeutic molecular targets to regulate ovarian function and treat ovarian disease. PMID- 19557762 TI - Glycosylphosphatidylinositols are potential targets for the development of novel inhibitors for aerolysin-type of pore-forming bacterial toxins. AB - Many bacteria produce toxins that cause damage through the formation of pores in the host cell membrane. Some of these toxins, such as aerolysin, use glycosylphosphatidylinositols (GPIs) as their binding receptors to assist the pore formation on the host cell surface and the subsequent insertion of the resultant pores into the cell membrane. GPIs are a class of complex glycolipids that anchor surface proteins and glycoproteins onto the cell membrane in eukaryotic species. This review has summarized the reported evidences supporting the GPI-dependent pore-forming mechanism for aerolysin-type of toxins and analyzed the possibility of targeting this unique process for the design and development of novel GPI-based inhibitors for these pore-forming bacterial toxins. PMID- 19557763 TI - Temporalis muscle flap for reconstruction of skull base defects. AB - BACKGROUND: The temporalis muscle flap (TMF) is a valuable reconstructive technique utilized in a variety of challenging defects. However, its use for repair of skull base defects is less commonly reported. METHODS: A retrospective chart review was conducted for 35 patients who underwent reconstruction of skull base defects between March 1999 and July 2006 at a tertiary referral hospital. Patients with skull base defects after trauma or extirpative surgery underwent reconstruction with a TMF. The measured outcomes were as follows: defect size/location, need for additional flaps, bone necrosis, hardware exposure, dehiscence, cerebrospinal fluid (CSF) leak, and meningitis. RESULTS: Forty-two patients underwent reconstruction with a TMF, and 35/42 patient records were available for review. No flap failures, 1 transient CSF leak, 3 hardware exposures distant from the temporalis recipient site, and 3 hydroxyapatite cement infections or foreign body reaction were observed. CONCLUSION: The TMF represents a versatile reconstructive technique employed with minimal morbidity and a low complication rate to repair defects of the skull base. PMID- 19557764 TI - Fourier transform convolution integrals applied to generalized Born molecular volume. AB - Generalized Born methods are currently among the solvation models most commonly used for biological applications. We reformulate the generalized Born molecular volume method initially described by (Lee et al, 2003, J Phys Chem, 116, 10606; Lee et al, 2003, J Comp Chem, 24, 1348) using fast Fourier transform convolution integrals. Changes in the initial method are discussed and analyzed. Finally, the method is extensively checked with snapshots from common molecular modeling applications: binding free energy computations and docking. Biologically relevant test systems are chosen, including 855-36091 atoms. It is clearly demonstrated that, precision-wise, the proposed method performs as good as the original, and could better benefit from hardware accelerated boards. PMID- 19557765 TI - Predicting the tautomeric equilibrium of acetylacetone in solution. I. The right answer for the wrong reason? AB - This study investigates how the various components (method, basis set, and treatment of solvent effects) of a theoretical approach influence the relative energies between keto and enol forms of acetylacetone, which is an important model system to study the solvent effects on chemical equilibria from experiment and theory. The computations show that the most popular density functional theory (DFT) approaches, such as B3LYP overestimate the stability of the enol form with respect to the keto form by approximately 10 kJ mol(-1), whereas the very promising SCS-MP2 approach is underestimating it. MP2 calculations indicate that in particular the basis set size is crucial. The Dunning Huzinaga double zeta basis (D95z(d,p)) used in previous studies overestimates the stability of the keto form considerably as does the popular split-valence plus polarization (SVP) basis. Bulk properties of the solvent included by continuum approaches strongly stabilize the keto form, but they are not sufficient to reproduce the reversal in stabilities measured by low-temperature nuclear magnetic resonance experiments in freonic solvents. Enthalpic and entropic effects further stabilize the keto form, however, the reversal is only obtained if also molecular effects are taken into account. Such molecular effects seem to influence only the energy difference between the keto and the enol forms. Trends arising due to variation in the dielectric constant of the solvent result from bulk properties of the solvent, i.e., are already nicely described by continuum approaches. As such this study delivers a deep insight into the abilities of various approaches to describe solvent effects on chemical equilibria. PMID- 19557767 TI - Neuronal activity rapidly induces transcription of the CREB-regulated microRNA 132, in vivo. AB - Activity-dependent changes in gene-expression are believed to underlie the molecular representation of memory. In this study, we report that in vivo activation of neurons rapidly induces the CREB-regulated microRNA miR-132. To determine if production of miR-132 is regulated by neuronal activity its expression in mouse brain was monitored by quantitative RT-PCR (RT-qPCR). Pilocarpine-induced seizures led to a robust, rapid, and transient increase in the primary transcript of miR-132 (pri-miR-132) followed by a subsequent rise in mature microRNA (miR-132). Activation of neurons in the hippocampus, olfactory bulb, and striatum by contextual fear conditioning, odor-exposure, and cocaine injection, respectively, also increased pri-miR-132. Induction kinetics of pri miR-132 were monitored and found to parallel those of immediate early genes, peaking at 45 min and returning to basal levels within 2 h of stimulation. Expression levels of primary and mature-miR-132 increased significantly between postnatal Days 10 and 24. We conclude that miR-132 is an activity-dependent microRNA in vivo, and may contribute to the long-lasting proteomic changes required for experience-dependent neuronal plasticity. PMID- 19557768 TI - Highly selective determination of phenolphthalein by flow injection chemiluminescence method based on a molecular imprinting polymer. AB - The phenolphthalein-imprinted polymer was prepared with methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linker. Taking advantage of the quenching effect of phenolphthalein on the potassium permanganate-HCl-anhydrous alcohol chemiluminescence system, a new model was established to determine phenolphthalein by a highly selective flow injection chemiluminescence method. The traditional flow-though cell was replaced with a novel flow path using a Y-shaped polymethyl methacrylate column, through which the three reactants were injected simultaneously. The linear range of this assay was from 1.0 x 10(-8) to 1.0 x 10(-6) g/mL (r = 0.9978). The limit of detection was 8.9 x 10(-9) g/mL. The relative standard deviation for the determination of 1.0 x 10(-8) g/mL phenolphthalein solution was below 2.9% (n = 11). The proposed method was applied to the determination of phenolphthalein in real samples with satisfactory results. PMID- 19557769 TI - Improved survival in red blood cell transfusion dependent patients with primary myelofibrosis (PMF) receiving iron chelation therapy. AB - Many patients with primary myelofibrosis (PMF) become red blood cell (RBC) transfusion dependent (TD), risking iron overload (IOL). Iron chelation therapy (ICT) may decrease the risk of haemosiderosis associated organ dysfunction, though its benefit in PMF is undefined. To assess the effect of TD and ICT on survival in PMF, we retrospectively reviewed 41 patients. Clinical data were collected from the database and by chart review. The median age at PMF diagnosis was 64 (range 43-86) years. Median white blood cell (WBC) count at diagnosis was 7.6 (range 1.2-70.9) x 10(9)/L; haemoglobin 104 (62-145) G/L; platelets 300 (38 2088) x 10(9)/L. Lille, Strasser, Mayo and International Prognostic System (IPS) scores were: low risk, n = 15, 8, 11, 3; intermediate, n = 15, 19, 9, 16; high, n = 5, 11, 5, 7; respectively. Primary PMF treatment was: supportive care, n = 23; hydroxyurea, n = 10; immunomodulatory, n = 4; splenectomy, n = 2. Sixteen patients were RBC transfusion independent (TI) and 25 TD; of these 10 received ICT for a median of 18.3 (0.1-117) months. Pre-ICT ferritin levels were a median of 2318 (range 263-8400) and at follow up 1571 (1005-3211 microg/L (p = 0.01). In an analysis of TD patients, factors significant for overall survival (OS) were: WBC count at diagnosis (p = 0.002); monocyte count (p = 0.0001); Mayo score (p = 0.05); IPS (p = 0.02); number of RBC units (NRBCU) transfused (p = 0.02) and ICT (p = 0.003). In a multivariate analysis, significant factors were: NRBCU (p = 0.001) and ICT (p = 0.0001). Five year OS for TI, TD-ICT and TD-NO ICT were: 100, 89 and 34%, respectively (p = 0.003). The hazard ratio (HR) for receiving >20 RBCU was 7.6 (95% Confidence Intervals [CI] 1.2-49.3) and for ICT was 0.15 (0.03 0.77). In conclusion, 61% of PMF patients developed RBC-TD which portended inferior OS; however patients receiving ICT had comparatively improved OS, suggesting a clinical benefit. Prospective studies of IOL and the impact of ICT in PMF are warranted. PMID- 19557770 TI - Lipid peroxidation in the liver of rats treated with V and/or Mg in drinking water. AB - The effect of V(5+) and Mg treatment on spontaneous and stimulated lipid peroxidation (LPO) was studied in liver supernatants obtained from outbred 5 month-old, albino male Wistar rats. The 2-month-old animals daily received deionized water to drink (control, group I); group II - water solution of NaVO(3) (SMV) at a concentration of 0.125 mg V ml(-1); group III - water solution of MgSO(4) (MS) at a concentration of 0.06 mg Mg ml(-1), group IV - water solution of SMV-MS at the same concentrations as in groups II and III for V and Mg, respectively, over a 12-week period. Three metal salts were selected as agents that may modify the LPO process (FeSO(4), NaVO(3) and MgSO(4)). V-intoxicated rats and those treated with V and Mg in combination had higher liver spontaneous malondialdehyde (MDA) formation, compared with the control and Mg-supplemented animals. In the same groups of animals the total antioxidant status (TAS) was also significantly lowered, in comparison with the control. In the supernatants obtained from the above-mentioned groups of rats a significant increase in MDA concentration was found in the presence of exogenous 30 microm FeSO(4) as well as 30, 100, 200 and 400 microm NaVO(3), compared with groups I and III. Significantly elevated MDA production was also observed in the supernatants obtained from the rats exposed to V and Mg in combination in the presence of exogenous 100 and 200 microm MgSO(4) in comparison with the control and group III as well as in the presence of exogenous 400 and 600 microm MgSO(4) compared only with group III. In vitro treatment with 1000 microm MgSO(4 )of control liver supernatants and those obtained from group III significantly enhanced MDA level, compared with spontaneous MDA formation. The two-way ANOVA indicated that the changes in the basal MDA level and in TAS in the rats at combined V and Mg application, were not due to V-Mg interaction, but resulted from independent action of V. In addition, the three-way ANOVA revealed that the changes in LPO induced by in vitro treatment of liver supernatants with exogenous Fe or V or Mg (600, 800 and 1000 microm) were a consequence of independent action of those metals and they also resulted from the interactions between Fe(exog) and V(end) and between V(end) and V(exog). In conclusion, V consumed by the rats with drinking water at a dose of 12 mg V kg(-1) body weight per 24 h for 12 weeks decreased TAS and enhanced spontaneous LPO in the hepatic tissue, which confirms its pro-oxidant potential, was also found in in vitro conditions with regard to LPO. Mg administered to rats in combination with V, at the concentration used, neither reduced nor intensified the basal LPO, compared with V-only treated animals; however, its stimulating effect on LPO was revealed in in vitro conditions, which requires further study. PMID- 19557772 TI - Diamond- and graphite-like octacyanometalate-based polymers induced by metal ions. AB - By using cyclohexane-1,2-diamine (chxn), Ni(ClO(4))(2) x 6 H(2)O and Na(3)[Mo(CN)(8)] x 4 H(2)O, a 3D diamond-like polymer {[Ni(II)(chxn)(2)](2)[Mo(IV)(CN)(8)] x 8 H(2)O}(n) (1) was synthesised, whereas the reaction of chxn and Cu(ClO(4))(2) x 6 H(2)O with Na(3)[M(V)(CN)(8)] x 4 H(2)O (M = Mo, W) afforded two isomorphous graphite-like complexes {[Cu(II)(chxn)(2)](3)[Mo(V)(CN)(8)](2) x 2 H(2)O}(n) (2) and {[Cu(II)(chxn)(2)](3)[W(V)(CN)(8)](2) x 2 H(2)O}(n) (3). When the same synthetic procedure was employed, but replacing Na(3)[Mo(CN)(8)] x 4 H(2)O by (Bu(3)NH)(3)[Mo(CN)(8)] x 4 H(2)O (Bu(3)N = tributylamine), {[Cu(II)(chxn)(2)Mo(IV)(CN)(8)][Cu(II)(chxn)(2)] x 2 H(2)O}(n) (4) was obtained. Single-crystal X-ray diffraction analyses showed that the framework of 4 is similar to 2 and 3, except that a discrete [Cu(chxn)(2)](2+) moiety in 4 possesses large channels of parallel adjacent layers. The experimental results showed that in this system, the diamond- or graphite-like framework was strongly influenced by the inducement of metal ions. The magnetic properties illustrate that the diamagnetic [Mo(IV)(CN)(8)] bridges mediate very weak antiferromagnetic coupling between the Ni(II) ions in 1, but lead to the paramagnetic behaviour in 4 because [Mo(IV)(CN)(8)] weakly coordinates to the Cu(II) ions. The magnetic investigations of 2 and 3 indicate the presence of ferromagnetic coupling between the Cu(II) and W(V)/Mo(V) ions, and the more diffuse 5d orbitals lead to a stronger magnetic coupling interaction between the W(V) and Cu(II) ions than between the Mo(V) and Cu(II) ions. PMID- 19557771 TI - Effect of lindane on CYP-mediated steroid hormone metabolism in male mice following in utero exposure. AB - A wide number of pesticides, including highly persistent organochlorinated compounds, such as lindane (LIN), may induce reproductive and developmental alterations by directly binding to the estrogen/androgen receptors or altering steroid hormone metabolism. In the present work, we have investigated whether LIN in utero exposure of CD1 mice affects the reproductive system in male offspring by causing an impairment of the CYP-dependent steroid hormone metabolism. Dam exposure to 25 mg kg(-1) b.w. LIN occurred during critical developmental periods, from gestational days 9 to 16. Effects on hepatic CYP-mediated testosterone (TST) hydroxylase, aromatase activities and testicular parameters were tested at postnatal days (PND 50, 65-69, 100) that are critical for sexual maturation in CD1 mice. In the adult F1 mice significant changes of male reproductive endpoints (testis weight, spermatid number) as well as dramatic effects on CYP-mediated TST metabolism were observed on PND 65-69, in the absence of any of systemic toxicity. The levels of TST 6beta- and 2alpha-hydroxylation and dehydrogenation showed the highest level of reduction, suggesting CYP 3A and 2C families as the major target of LIN induced effects. All changes were almost recovered on PND 100. No effects on aromatase activity were evidenced. Overall, these findings provide useful information for a better characterization of the LIN mode of action. They suggest that LIN-induced toxicity in males is linked to an impairment of steroid hormone homeostasis, due to CYP-mediated TST catabolism modulation and differs from LIN receptor-mediated mechanism previously reported in females. PMID- 19557773 TI - Matching and mismatching effects of hybrid chiral biaxial bisphosphine ligands in enantioselective hydrogenation of ketoesters. PMID- 19557775 TI - Stanislao Cannizzaro and the development of chemistry in Palermo from 1862 to 1871. AB - Stanislao Cannizzaro worked at Palermo University for about ten years. There he managed to establish a modern and well-equipped chemical laboratory. His international fame attracted co-workers even from abroad: Naquet, Lieben, and Korner came to Palermo to work with him. This greatly improved the quality of teaching and of research in Palermo, which became a worldwide acknowledged center of chemical culture. In Palermo, atomic-molecular theory was extensively taught and research was carried out on chemical atomicity (valence) and spatial structures of both aliphatic and aromatic organic molecules: Korner found the relative positions of substituent groups on the benzene ring, Paterno proved the equivalence of the four positions around each carbon atom. Thanks to Cannizzaro's and Paterno's efforts, a new journal, Gazzetta Chimica Italiana, was printed in Palermo. The journal collected original papers from Italian chemists, and summaries or translations of foreign ones. When Cannizzaro moved to Rome in 1872, Emanuele Paterno, the youngest of his pupils, took his place. Eventually, Paterno also moved to Rome, and the leading role of Palermo in Italian chemistry faded. PMID- 19557774 TI - Noncovalent modulation of pH-dependent reactivity of a Mn-salen cofactor in myoglobin with hydrogen peroxide. AB - To demonstrate protein modulation of metal-cofactor reactivity through noncovalent interactions, pH-dependent sulfoxidation and 2,2'-azino-bis(3 ethylbenzthiazoline-6-sulphonic acid) (ABTS) oxidation reactivity of a designed myoglobin (Mb) containing non-native Mn-salen complex (1) was investigated using H2O2 as the oxidant. Incorporation of 1 inside the Mb resulted in an increase in the turnover numbers through exclusion of water from the metal complex and prevention of Mn-salen dimer formation. Interestingly, the presence of protein in itself is not enough to confer the increase activity as mutation of the distal His64 in Mb to Phe to remove hydrogen-bonding interactions resulted in no increase in the turnover numbers, while mutation His64 to Arg, another residue with ability to hydrogen-bond interactions, resulted in an increase in reactivity. These results strongly suggest that the distal ligand His64, through its hydrogen-bonding interaction, plays important roles in enhancing and fine tuning reactivity of the Mn-salen complex. Nonlinear least-squares fitting of rate versus pH plots demonstrates that 1.Mb(H64X) (X=H, R and F) and the control Mn-salen 1 exhibit pKa values varying from pH 6.4 to 8.3, and that the lower pKa of the distal ligand in 1.Mb(H64X), the higher the reactivity it achieves. Moreover, in addition to the pKa at high pH, 1.Mb displays another pKa at low pH, with pKa of 5.0+/-0.08. A comparison of the effect of different pH on sulfoxidation and ABTS oxidation indicates that, while the intermediate produced at low pH conditions could only perform sulfoxidation, the intermediate at high pH could oxidize both sulfoxides and ABTS. Such a fine-control of reactivity through hydrogen-bonding interactions by the distal ligand to bind, orient and activate H2O2 is very important for designing artificial enzymes with dramatic different and tunable reactivity from catalysts without protein scaffolds. PMID- 19557776 TI - Photophysics of an intramolecular hydrogen-evolving Ru-Pd photocatalyst. AB - Photoinduced electron-transfer processes within a precatalyst for intramolecular hydrogen evolution [(tbbpy)(2)Ru(tpphz)PdCl(2)](2+) (RuPd; tbbpy = 4,4'-di-tert butyl-2,2'-bipyridine, tpphz = tetrapyrido[3,2-a:2',3'c:3'',2'',-h:2''',3''' j]phenazine) have been studied by resonance Raman and ultrafast time-resolved absorption spectroscopy. By comparing the photophysics of the [(tbbpy)(2)Ru(tpphz)](2+) subunit Ru with that of the supramolecular catalyst RuPd, the individual electron-transfer steps are assigned to kinetic components, and their dependence on solvent is discussed. The resonance Raman data reveal that the initial excitation of the molecular ensemble is spread over the terminal tbbpy and the tpphz ligands. The subsequent excited-state relaxation of both Ru and RuPd on the picosecond timescale involves formation of the phenazine-centered intraligand charge-transfer state, which in RuPd precedes formation of the Pd reduced state. The photoreaction in the heterodinuclear supramolecular complex is completed on a subnanosecond timescale. Taken together, the data indicate that mechanistic investigations must focus on potential rate-determining steps other than electron transfer between the photoactive center and the Pd unit. Furthermore, structural variations should be directed towards increasing the directionality of electron transfer and the stability of the charge-separated states. PMID- 19557777 TI - Enantioselective organocatalytic domino oxa-Michael/aldol/hemiacetalization: synthesis of polysubstituted furofuranes containing four stereocenters. PMID- 19557778 TI - Probing dynamics within amyloid fibrils using a novel capping method. PMID- 19557779 TI - Chirality relay to access oxygenated angular aromatic polyketides. PMID- 19557780 TI - Use of gel-assembly to fabricate multi-component molecular gradient layers and the investigation of structure and electron transport therein. PMID- 19557781 TI - Gold-loaded carbon nanoparticles from poly(vinyl alcohol)-b-poly(acrylonitrile) non-shell-cross-linked micelles. AB - Herein we show that a new amphiphilic poly(vinyl alcohol)-b-poly(acrylonitrile) block copolymer dispersed in water can be easily loaded with gold nanoparticles by addition of chlorauric acid followed by reduction by sodium borohydride. After deposition of the so-loaded micelles onto a silicon wafer, followed by an appropriate thermal treatment, the poly(acrylonitrile) core of the micelles is carbonized, while the poly(vinyl alcohol) shell is completely decomposed and volatilized, leading to gold encapsulated in carbon nanoparticles. The morphology of the micelles is maintained during thermal treatment without requiring shell cross-linking of the micelles prior to pyrolysis. PMID- 19557782 TI - Nickel-catalyzed three-component [2+2+2] cycloaddition reaction of arynes, alkenes, and alkynes. PMID- 19557784 TI - Relative stability of closo-closo, closo-nido, and nido-nido macropolyhedral boranes: the role of orbital compatibility. AB - The concept of orbital compatibility is used to explain the relative energies of different macropolyhedral structural patterns such as closo-closo, closo-nido, and nido-nido. A large polyhedral borane condenses preferentially with a smaller polyhedron owing to orbital compatibility. Calculations carried out at the B3LYP/6-31G* level show that the macropolyhedron closo(12)-closo(6) is the most preferred structural pattern among the face-sharing closo-closo systems. The relative stabilities of four-shared-atom closo-closo, three-shared-atom closo closo, three-shared-atom closo-nido, edge-sharing closo-nido, and edge-sharing nido-nido structures are in accordance with the difference in the number of vertices of the individual polyhedra of the macropolyhedra. When the difference in the number of vertices of the individual polyhedra is large, the stability of the macropolyhedra is also large. Calculations further show that the orbital compatibility plays an important role in deciding the stability of the macropolyhedral boranes with more than two polyhedral units. The dependence of the orbital compatibility on the relative stability of the macropolyhedron varies with other factors such as inherent stability of the individual polyhedron and steric factors. PMID- 19557783 TI - A cytotoxic ruthenium tris(bipyridyl) complex that accumulates at plasma membranes. PMID- 19557785 TI - A DBFOX-Ph-based combinatorial catalyst for enantioselective fluorination of aryl acetyl and 3-butenoyl thiazolidinones. PMID- 19557786 TI - Umpolung of electrophilic terminal phosphinidene complexes by interaction with nucleophilic carbenes. PMID- 19557787 TI - The performance of dental pulp stem cells on nanofibrous PCL/gelatin/nHA scaffolds. AB - The aim of current study is to investigate the in vitro and in vivo behavior of dental pulp stem cells (DPSCs) seeded on electrospun poly(epsilon-caprolactone) (PCL)/gelatin scaffolds with or without the addition of nano-hydroxyapatite (nHA). For the in vitro evaluation, DNA content, alkaline phosphatase (ALP) activity and osteocalcin (OC) measurement showed that the scaffolds supported DPSC adhesion, proliferation, and odontoblastic differentiation. Moreover, the presence of nHA upregulated ALP activity and promoted OC expression. Real-time PCR data confirmed these results. SEM micrographs qualitatively confirmed the proliferation and mineralization characteristics of DPSCs on both scaffolds. Subsequently, both scaffolds seeded with DPSCs were subcutaneously implanted into immunocompromised nude mice. Scaffolds with nHA but without cells were implanted as control. Histological evaluation revealed that all implants were surrounded by a thin fibrous tissue capsule without any adverse effects. The cell/scaffold composites showed obvious in vivo hard tissue formation, but there was no sign of tissue ingrowth. Further, the combination of nHA in scaffolds did upregulate the expression of specific odontogenic genes. In conclusion, the incorporation of nHA in nanofibers indeed enhanced DPSCs differentiation towards an odontoblast-like phenotype in vitro and in vivo. PMID- 19557788 TI - Biological relevance of ion energy in performance of human endothelial cells on ion-implanted flexible polyurethane surfaces. AB - To improve the biocompatibility of polyurethane (PUR), we modified the surface by irradiation with different ions (Carbon; C, Oxygen; O, Nitrogen; N, or Argon; Ar) at 0.3-50 keV energy and doses of 1,00E+13 - 1,00E+15 ions/cm(2). The effects of ion implantation using different ion energies and densities were observed on adhesion, proliferation, and viability of human umbilical vein endothelial cells (HUVECs). The long-term in vitro stability of ion-implanted PUR was also investigated. Ion irradiation moderately affected the surface roughness (R(a)), but strongly enhanced the work of adhesion (W(a)). Cell adhesion was markedly improved on O-, N-, and Ar-, but not on C-implanted PUR surfaces. Medium ion energies and lower ion doses produced the best HUVEC attachment and proliferation, indicating the importance of choosing the proper range of energy applied during ion irradiation. In addition, apoptosis rates were significantly reduced when compared with unmodified PUR (uPUR). N implantation significantly protected the surface, although C implantation led to stronger surface erosions than on uPUR. In total, ion implantation on flexible PUR surfaces strongly improved the material surface characteristics and biocompatibility. Electron beam ion implantation within an appropriate energy window is thus a key to improving flexible PUR surfaces for clinical use to support endothelial cell performance. Thus, it can contribute to designing small-diameter grafts, which are in great demand, towards vascular tissue engineering applications. PMID- 19557789 TI - Polymeric matrix for drug delivery: honokiol-loaded PCL-PEG-PCL nanoparticles in PEG-PCL-PEG thermosensitive hydrogel. AB - In this article, we demonstrated a novel injectable polymer matrix: honokiol (HK) loaded poly (epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon caprolactone) (PCL-PEG-PCL, PCEC) nanoparticles in thermosensitive poly(ethylene glycol)-poly(epsilon-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) hydrogel for the drug local delivery. First, HK, as a model hydrophobic drug, was loaded into PCL-PEG-PCL nanoparticles by emulsion solvent evaporation method to overcome its poor water solubility. Then, the HK-loaded PCEC nanoparticles (HK PCEC) were incorporated into thermosensitive PEG-PCL-PEG hydrogel, which was sol at low temperature and could gel as a depot for sustained release of drug in situ after topical injection. The HK-PCEC incorporated PECE hydrogel (HK-PCEC-PECE) was biodegradable and could be gradually eliminated from the injection site in about 2 weeks after subcutaneously injected into mice. The in vitro release studies indicated that HK could be released from HK-PCEC and HK-PCEC-PECE in a sustained manner. Such biodegradable smart drug-delivery system might have great potential application in injectable hydrophobic drug local delivery system. PMID- 19557790 TI - Biomechanical and biologic effects of meniscus stabilization using triglycidyl amine. AB - The susceptibility of meniscus allografts to enzymatic degradation may be reduced through tissue stabilization. We have previously reported on an epoxide-based crosslinker, triglycidyl amine (TGA), which can be used alone or with a bisphosphonate (MABP) to stabilize heterograft heart valves and reduce their pathologic calcification. Our objective was to evaluate the effects of TGA and TGA-MABP pretreatment on an orthopedic allograft involving meniscus crosslinking, degradation, calcification, and compressive properties. Ovine menisci treated with TGA or TGA-MABP for up to seven days and glutaraldehyde crosslinked controls were examined in vitro for degree of crosslinking, resistance to degradation by collagenase, and material property changes. Likewise treated menisci were implanted in rats for eight weeks and examined for calcium content and biomechanical changes. TGA treatment for three days significantly reduced collagen loss by 88% and increased thermal denaturation temperatures (Ts) above 80 degrees C versus Ts of 70 degrees C or less for non-crosslinked meniscus. In vitro, TGA and TGA-MABP significantly increased aggregate modulus by 19% and 32% compared to native controls, respectively. TGA decreased permeability by 53% while TGA-MABP increased it by 303%. In vivo, TGA significantly reduced explant calcification by 42% compared to glutaraldehyde, and including MABP reduced it by 90%. Analyses revealed that TGA and TGA-MABP stabilized menisci had significantly lower modulus and permeability values than glutaraldehyde controls by at least 28% and 86%, respectively. It is concluded that TGA crosslinking of meniscus increases resistance to both collagenase degradation and pathologic calcification, while demonstrating comparable or improved biomechanical properties versus glutaraldehyde controls. PMID- 19557791 TI - Glass ionomer cement hardness after different materials for surface protection. AB - The aim of the present study was to evaluate the Knoop hardness of high viscous glass ionomer cement (GIC) Ketac Molar Easy Mix (3M ESPE, Saint Paul, USA) submitted to different types of compounds for surface protection. Sixty specimens of GIC were made in PVC molds with 7.5 mm diameter and 2.5 mm thickness. Divided in 6 groups: G1, Control (no protection); G2, Cavitine (Copal varnish); G3, Magic bond (Adhesive); G4, Adper Single Bond 2 (Single bottle adhesive); G5, Solid Petroleum Jelly; G6, Nail Varnish. The surface protection was applied after initial setting reaction. The specimens were immersed in deionized water, at 37 degrees C, for 24 h. The surfaces were polished in a rotation machine (Aropol 2V). The hardness test was accomplished in a Digital Microhardness tester HVS 100. In each specimen five indentations were done and repeated after 30 days and 4 months, under the same conditions. The results were submitted to Two-way ANOVA and Tukey Test. The only material that differed from the control group was the nail varnish (p < 0.001), with the other materials showing no significant difference from the control group. It was concluded that the best material for surface protection of GIC was the nail varnish, but because of possible harmful effects, petroleum jelly could be a better option. PMID- 19557792 TI - In vivo response to cross-linked polyethylene and polycarbonate-urethane particles. AB - This study was undertaken to examine macrophage response to polycarbonate urethane, a proposed alternative material to polyethylene in acetabular components of total hip arthroplasty. Polyethylene wear debris from total joint replacements has been linked to osteolysis and implant lifespan. It has been shown in vitro, that polyethylene particles cleaned of endotoxin generate less of an inflammatory cytokine response than endotoxin bound particles. Comparative particle induced effects on implant fixation were tested using endotoxin free cross-linked ultra-high molecular weight polyethylene (x-UHMWPE) and polycarbonate-urethane (PCU) particles with and without intraperitoneal injection (IP) of lipopolysaccharide (LPS) using a Ti-alloy femoral intramedullary nail rat model. MicroCT and mechanical testing assessment of peri-implant bone indicated significantly less bone and lower fixation strength, respectively, when the implant was surrounded by xUHMWPE particles compared to PCU particles (with and without LPS IP). This indicates particles of PCU may be less disruptive to bone implant fixation than x-UHMWPE in vivo, under both LPS free and challenged conditions. PMID- 19557793 TI - Polyunsaturated fatty acids inhibit PI3K activity in a yeast-based model system. AB - The phosphatidylinositol 3-kinase (PI3K) pathway controls the regulation of cell growth, proliferation, migration and apoptosis. In many tumors, the PI3K gene is mutated or overexpressed, and/or the PI3K pathway is hyperactive. PI3K is therefore a potential pharmacological target for the development of anti-tumor drugs. Some polyunsaturated fatty acids (PUFA), when given in the diet, may lead to a decrease in PI3K activity. We used a yeast-based model to reconstitute the PI3K/PTEN/Akt pathway to study the effects of long-chain polyunsaturated n-3 fatty acids on PI3K, and found that various PUFA were able to alleviate toxicity induced by overexpression of PI3K. The various PUFA had no significant effect on the steady-state level of PI3K catalytic subunit proteins (p110alpha) in yeast. However, depletion of phosphatidylinositol 4,5-bisphosphate due to overexpression of the p110alpha subunit was significantly reduced by treating the yeast cells with the various PUFA. The inhibition of mammalian PI3K, expressed in an exogenous cellular context in yeast, is likely to be a direct effect of these PUFA on PI3K rather than on other mammalian endogenous or environmental factors. These results are particularly promising given the abundance of active PUFA in marine foodstuffs and especially fish oils. PMID- 19557794 TI - Multi-sensor plant imaging: Towards the development of a stress-catalogue. AB - Agricultural production is limited by a wide range of abiotic (e.g. drought, waterlogging) and biotic (pests, diseases and weeds) stresses. The impact of these stresses can be minimized by appropriate management actions such as irrigation or chemical pesticide application. However, further optimization requires the ability to diagnose and quantify the different stresses at an early stage. Particularly valuable information of plant stress responses is provided by plant imaging, i.e. non-contact sensing with spatial resolving power: (i) thermal imaging, detecting changes in transpiration rate and (ii) fluorescence imaging monitoring alterations in photosynthesis and other physiological processes. These can be supplemented by conventional video imagery for study of growth. An efficient early warning system would need to discriminate between different stressors. Given the wide range of sensors, and the association of specific plant physiological responses with changes at particular wavelengths, this goal seems within reach. This is based on the organization of the individual sensor results in a matrix that identifies specific signatures for multiple stress types. In this report, we first review the diagnostic effectiveness of different individual imaging techniques and then extend this to the multi-sensor stress-identification approach. PMID- 19557795 TI - Cloning and overexpression of a thermostable signal peptide peptidase (SppA) from Thermoplasma volcanium GSS1 in E. coli. AB - In this study, a gene coding for thermophilic serine protease of the ClpP class from the thermoacidophilic archaeon Thermoplasma volcanium (Tpv) was cloned and expressed in Escherichia coli. The primary sequence and domain analysis of this enzyme showed similarities (50-60% similarity) to signal peptide peptidases (SppA) of bacteria and other archaea. An increase of about tenfold in the activity was achieved by overexpression of Tpv SppA in E. coli, as detected by enzyme assays conducted using Ala-Ala-Phe-pNa and N-Suc-Ala-Ala-Pro-Phe-pNA as substrates. The recombinant enzyme, purified using an anion exchange column chromatography, displayed an apparent molecular mass of 26 kDa on SDS-PAGE analysis. Purified Tpv SppA was active in a broad range of pH and temperature with maximal activity at 60 degrees C and between pH 7.5 and pH 8.0. The activity of the enzyme was strongly inhibited by inhibitors typical for serine proteases, i.e., chymostatin and PMSF. The activity of the Tpv SppA and the stability at high temperature were significantly enhanced in the presence of 5 mM Ca(2+) ions. Our multiple sequence alignment data revealed a conserved Ser/Lys catalytic dyad in Tpv SppA that comprised Ser76 (nucleophile) and Lys128 (general base) residues. A search for a transmembrane domain using automated programs did not predict any signal peptide associated with the Tpv SppA and, therefore, suggested a cytoplasmic location for this enzyme. PMID- 19557796 TI - Application of a PEG/salt aqueous two-phase partition system for the recovery of monoclonal antibodies from unclarified transgenic tobacco extract. AB - Aqueous two-phase partition systems (ATPS) have been widely used for the separation of a large variety of biomolecules. In the present report, the application of a polyethylene glycol/phosphate (PEG/phosphate) ATPS for the separation of anti-HIV monoclonal antibodies 2G12 (mAb 2G12) and 4E10 (mAb 4E10) from unclarified transgenic tobacco crude extract was investigated. Optimal conditions that favor opposite phase partitioning of plant debris/mAb as well as high recovery and purification were found to be 13.1% w/w (PEG 1500), 12.5% w/w (phosphate) at pH 5 with a phase ratio of 1.3 and 8.25% w/w unclarified tobacco extract load. Under these conditions, mAb 2G12 and mAb 4E10 were partitioned at the bottom phosphate phase with 85 and 84% yield and 2.4- and 2.1-fold purification, respectively. The proposed ATPS was successfully integrated in an affinity-based purification protocol, using Protein A, yielding antibodies of high purity and yield. In this study, ATPS was shown to be suitable for initial protein recovery and partial purification of mAb from unclarified transgenic tobacco crude extract. PMID- 19557797 TI - Improved donor/acceptor BRET couples for monitoring beta-arrestin recruitment to G protein-coupled receptors. AB - We report highly sensitive bioluminescence resonance energy transfer (BRET) assays with optimized donor/acceptor couples. We combined the energy donors Renilla luciferase (Rluc) and the Rluc8 variant with the energy acceptors yellow fluorescent protein, the YPet variant and the Renilla green fluorescent protein (RGFP). Different donor/acceptor couples were tested in well-established assays measuring ligand-induced beta-arrestin (betaARR) intramolecular rearrangements and recruitment to G protein-coupled receptors. We show increased sensitivity with Rluc8/YPet and Rluc8/RGFP couples and measured previously undetectable BRET signals. These tools improve existing betaARR assays and offer new options for the development of future BRET assays. PMID- 19557798 TI - Metals sorption from aqueous solutions by Kluyveromyces marxianus: process optimization, equilibrium modeling and chemical characterization. AB - The dead Kluyveromyces marxianus biomass, a fermentation industry waste, was used to explore its sorption potential for lead, mercury, arsenic, cobalt, and cadmium as a function of pH, biosorbent dosage, contact time, agitation speed, and initial metal concentration. The equilibrium data fitted the Langmuir model better for cobalt and cadmium, but Freundlich isotherm for all metals tested. At equilibrium, the maximum uptake capacity (Qmax) was highest for lead followed by mercury, arsenic, cobalt, and cadmium. The RL values ranged between 0-1, indicating favorable sorption of all test metals by the biosorbent. The maximum Kf value of Pb showed its efficient removal from the solution. However, multi metal analysis depicted that sorption of all metals decreased except Pb. The potentiometric titration of biosorbent revealed the presence of functional groups viz. amines, carboxylic acids, phosphates, and sulfhydryl group involved in heavy metal sorption. The extent of contribution of functional groups and lipids to biosorption was in the order: carboxylic>lipids>amines>phosphates. Blocking of sulfhydryl group did not have any significant effect on metal sorption. PMID- 19557799 TI - Industrial production of enzyme-modified wool fibers for machine-washable bed coverings. AB - Enzyme technology is explored on wool fibers to prevent shrinkage and consolidation behavior during washing of woolen bed coverings using normal household machine conditions. Enzyme modification of wool fibers after two different pretreatments has been realized on industrial scale. Enlarged proteolytic enzyme by chemical modification was applied successfully to prevent substantial fiber strength loss. Felt-ball analysis of the fibers as obtained from this industrial process showed substantial improvement in felting resistance. Further processing of these enzyme-modified fibers and finally integration in bed covering quilts have been executed successfully on industrial production lines. The observed fiber losses during processing were in the range of 4.5-6% which is comparable with that of nonmodified fibers. The machine washability of these produced bed covering quilts was tested in a household washing machine using both wool and normal wash programs applied at different temperatures. It appeared that, contrary to the good washing results in terms of shrinkage and consolidation resistance using the wool program at moderate temperatures, this resistance is marginal when washed with the normal washing program with higher mechanical agitation level or with the wool program at elevated temperature. This result was different from that obtained with woolen fabrics and explained by the less-structured organisation of fibers within a fleece. PMID- 19557800 TI - A putative role for human BFK in DNA damage-induced apoptosis. AB - Human BFK (BCL-2 family kin) is a novel pro-apoptotic BCL-2 family member specifically expressed in the gastrointestinal tract. BFK has the characteristic BH3 domain, which was shown to be essential for the apoptosis-inducing activity of pro-apoptotic BCL-2 family members. When overexpressed, BFK interacts with BCL XL and BCL-W but not BCL-2 or BAD in co-immunoprecipitations studies. We find that BFK exhibits striking similarity to BID in the way it is activated through cleavage during apoptosis. The endogenous and cleaved versions of BFK are readily recognized by the rabbit and mouse sera raised against human BFK. An ideal caspase 3 or 7 target sequence, DEVD (amino acids 38-41), is evident N-terminal to the BH3 domain. A recombinant version of the protein containing all residues downstream of the putative caspase cleavage site induces apoptosis in human colon cancer cells, HCT116, and in wild-type mouse embryonic fibroblasts (MEFs), which can be reversed by co-expression of BCL-XL or BCL-W. BFK becomes activated through caspase-dependent cleavage during DNA damage-induced apoptosis. The cleaved form of the protein is dependent on the presence of BAX or BAK for its ability to induce apoptosis, since BAX(-/-)-BAK(-/-) double-knockout MEFs are completely resistant to BFK-induced apoptosis. PMID- 19557802 TI - Synthesis and evaluation of 1-(1-(Benzo[b]thiophen-2-yl)cyclohexyl)piperidine (BTCP) analogues as inhibitors of trypanothione reductase. AB - Thirty two analogues of phencyclidine were synthesised and tested as inhibitors of trypanothione reductase (TryR), a potential drug target in trypanosome and leishmania parasites. The lead compound BTCP (1, 1-(1-benzo[b]thiophen-2-yl cyclohexyl) piperidine) was found to be a competitive inhibitor of the enzyme (K(i)=1 microM) and biologically active against bloodstream T. brucei (EC(50)=10 microM), but with poor selectivity against mammalian MRC5 cells (EC(50)=29 microM). Analogues with improved enzymatic and biological activity were obtained. The structure-activity relationships of this novel series are discussed. PMID- 19557803 TI - Discovery of ligands for the human intestinal Di-/Tripeptide transporter (hPEPT1) using a QSAR-assisted virtual screening strategy. AB - The discovery of novel ligands for the hPEPT1 transporter is reported. By exploiting a fast and rigorously validated QSAR model in combination with the distance in activity-centered chemical space (DACCS) approach, a database of commercially available compounds (Sigma-Aldrich) was screened for virtual hits. Twelve compounds were then purchased and characterized in an apical [14C]Gly-Sar uptake competition assay. Four compounds displayed affinity in the medium-to-high range. A simple benzophenone derivative displayed high affinity with a sub millimolar binding constant (Ki=0.24 mM). The results of this study will serve as starting points for future projects, including the design and synthesis of compound libraries that seek to systematically explore the fundamental requirements for binding and transport by hPEPT1. PMID- 19557801 TI - Improved tricyclic inhibitors of trypanothione reductase by screening and chemical synthesis. AB - Trypanothione reductase (TryR) is a key validated enzyme in the trypanothione based redox metabolism of pathogenic trypanosomes and leishmania parasites. This system is absent in humans, being replaced with glutathione and glutathione reductase, and as such offers a target for selective inhibition. As part of a program to discover antiparasitic drugs, the LOPAC1280 library of 1266 compounds was screened against TryR and the top hits evaluated against glutathione reductase and T. brucei parasites. The top hits included a number of known tricyclic neuroleptic drugs along with other new scaffolds for TryR. Three novel druglike hits were identified and SAR studies on one of these using information from the tricyclic neuroleptic agents led to the discovery of a competitive inhibitor (K(i)=330 nM) with an improved potency against T. brucei (EC(50)=775 nM). PMID- 19557804 TI - Indoloquinolizidine-peptide hybrids as multiple agonists for D1 and D2 dopamine receptors. AB - Multiple-specificity ligands are considered promising pharmacological tools that may show higher efficacy in the treatment of diseases for which the modulation of a single target is therapeutically inadequate. We prepared a set of novel ligands for D1 and D2 dopamine receptors by combining two indolo[2,3-a]quinolizidine scaffolds with various tripeptide moieties. The binding and functional properties of these molecules were determined by radioligand binding studies in brain striatum membranes and by intracellular cAMP production assays in cells expressing different dopamine receptor subtypes. Some indoloquinolizidine-peptide hybrids, mainly with the trans configuration, showed dual agonist activity at both D1 and D2 dopamine receptors and may therefore be useful for testing the therapeutic potential of multivalent drugs on these targets. PMID- 19557805 TI - MRI-guided cryoablation: In vivo assessment of focal canine prostate cryolesions. AB - PURPOSE: To analyze the appearance of acute and chronic canine prostate cryolesions on T1-weighted (T1w) and T2-weighted (T2w) magnetic resonance imaging (MRI) and compare them with contrast-enhanced (CE) MRI and histology for a variety of freezing protocols. MATERIALS AND METHODS: Three different freezing protocols were used in canine prostate cryoablation experiments. Six acute and seven chronic (survival times ranging between 4-53 days) experiments were performed. The change in T2w signal intensity was correlated with freezing protocol parameters. The lesion area on T2w MRI was compared to CE-MRI. Histopathologic evaluation of the cryolesions was performed and visually compared to the appearance on MRI. RESULTS: The T2w signal increased from pre- to postfreeze at the site of the cryolesion, and the enhancement was higher for smaller freeze area and duration. The T2w lesion area was between the CE nonperfused area and the hyperenhancing CE rim. The appearance of the lesion on T1w and T2w imaging over time correlated with outcome on pathology. CONCLUSION: T1w and T2w MRI can potentially be used to assess cryolesions and to monitor tissue response over time following cryoablation. PMID- 19557806 TI - DCE-MRI pixel-by-pixel quantitative curve pattern analysis and its application to osteosarcoma. AB - PURPOSE: To present a novel curve pattern analysis (CPA) method to characterize and quantify signal curves from the dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) data without any prerequisites such as arterial input function (AIF) or T(1) measurement. MATERIALS AND METHODS: CPA parameters represent characteristics of the scaled DCE signal curve. Simulations were performed to investigate the dependence of CPA parameters on T(1), TR, and flip angle. In vivo studies were performed on five pediatric patients with osteosarcoma. Parametric maps were generated using the CPA method and a pharmacokinetic model-based method for comparison. RESULTS: Simulations show that CPA parameters varied less than 2% when T(1) changed from 300 msec to 1500 msec, and less than 10% when the flip angle changed from 30 degrees to 40 degrees. Various curve patterns can be qualitatively identified and recognized from CPA parameter maps. Simulation and in vivo studies showed that the CPA parameter had a strong correlation with k(ep), with correlation coefficients of 0.9983 in the simulation and 0.95 in the in vivo studies. CONCLUSION: A novel CPA method is presented. Simulations and in vivo studies showed that the CPA method provides a feasible alternative to quantifying DCE-MRI studies with possibly higher repeatability by minimizing variations potentially induced by AIF and T(1) estimations and model dependence. PMID- 19557807 TI - Effect of polymerization conditions on morphology and chromatographic characteristics of polyacrylamide-based beds (monoliths) for capillary electrochromatography and capillary liquid chromatography. AB - Pioneering research work on continuous or monolithic beds for LC was published in 1989. However, till date there is a lack of complex investigation of various factors affecting the morphology of the beds polymerized from water-soluble co monomers. In this article the effect of buffer, used as polymerization medium, concentration and pH, and total monomer concentration on the morphology of the resultant continuous beds are discussed. Evaluation of the continuous beds was performed by means of capillary LC, CEC, and inverse size-exclusion chromatography. PMID- 19557808 TI - Preparative separation and purification of liensinine, isoliensinine and neferine from seed embryo of Nelumbo nucifera GAERTN using high-speed counter-current chromatography. AB - A preparative high-speed counter-current chromatography method for separation and purification of liensinine, isoliensinine and neferine from seed embryo of Nelumbo nucifera GAERTN was successfully established by using n-hexane-ethyl acetate-methanol-water (5:8:4:5, v/v, containing 0.5% NH(4)OH) as the two-phase solvent system. From 200 mg of crude extract, 18.4 mg of liensinine, 19.6 mg of isoliensinine and 58.4 mg of neferine were obtained with the purity of 96.8, 95.9, and 98.6%, respectively. The identification of the three alkaloids was performed with (1)H NMR and (13)C NMR. PMID- 19557809 TI - New mathematic model for predicting chiral separation using molecular docking: mechanism of chiral recognition of triadimenol analogues. AB - The purpose of this paper was to study the enantioseparation mechanism of triadimenol compounds by carboxymethylated (CM)-beta-CD mediated CE. All the enantiomers were separated under the same experimental conditions to study the chiral recognition mechanism using a 30 mM sodium dihydrogen phosphate buffer at pH 2.2 adjusted by phosphoric acid. The inclusion courses between CM-beta-CD and enantiomers were investigated by the means of molecular docking technique. It was found that there were at least three points (one hydrophobic bond and two hydrogen bonds) involved in the interaction of each enantiomer with the chiral selectors. A new mathematic model has been built up based on the results of molecular mechanics calculations, which could analyze the relationship between the resolution of enantioseparation and the interaction energy in the docking area. Comparing the results of the separation by CE, the established mathematic model demonstrated good capability to predict chiral separation of triadimenol enantiomers using CM-beta-CD mediated CE. PMID- 19557810 TI - Preparative isolation and purification of antioxidative stilbene oligomers from Vitis chunganeniss using high-speed counter-current chromatography in stepwise elution mode. AB - Preparative high-speed counter-current chromatography (HSCCC) was successfully applied to the isolation and purification of three stilbene oligomers from Vitis chunganeniss using stepwise elution with a pair of two-phase solvent systems composed of n-hexane-ethyl acetate-methanol-water at (2:5:2:5, v/v) and (1:2:1:2, v/v). The preparative HSCCC separation was performed on 800 mg of crude sample yielding hopeaphenol (21.1 mg), amurensin G (37.2 mg) and vitisin A (95.6 mg) in a one-step separation, with purities over 95% as determined by HPLC. The structures of these three compounds were identified by MS, (1)H NMR and (13)C NMR. In addition, their antioxidant activities were screened by DPPH assay, where vitisin A showed strong antioxidant activity. Further EPR experiments with spin trapping technique demonstrated that vitisin A is a potent and selective singlet oxygen quencher, which may be used in singlet oxygen-mediated diseases as a pharmacological agent. PMID- 19557811 TI - The lipophilicity of parabens estimated on reverse phases chemically bonded and oil-impregnated plates and calculated using different computation methods. AB - The chromatographic behaviour of the parabens has been investigated on RP 18F(254S), RP-18WF(254S), CNF(254S), Diol F(254S) and silica gel 60F(254) plates impregnated with different oils (paraffin, olive, sunflower and corn) using methanol-water mixtures in different volume proportions as mobile phases, the regression determination coefficients being excellent (higher than 0.98 for the majority of compounds). Moreover, highly significant correlations were obtained between different experimental indices of lipophilicity (R(M0), b and scores corresponding to the first principal component (PC1)) and computed log P values. All types of stationary phases investigated appear to be highly suited for estimating the lipophilicity of the parabens. PMID- 19557812 TI - 1 mm ID poly(styrene-co-divinylbenzene) monolithic columns for high-peak capacity one- and two-dimensional liquid chromatographic separations of intact proteins. AB - The LC performance of a 1x50 mm polymer monolithic column format was demonstrated with high-peak capacity one- (1D) and offline two dimensional (2D) LC separations of intact proteins. After optimizing the RP 1D-LC conditions, including column temperature, flow rate and gradient time, a peak capacity of 475 was achieved within a 2-h analysis. The suitability of the monolithic column was also demonstrated for fast 1 min protein separations yielding 1 s peak widths determined at half peak height. In addition, an offline 2D-LC method was developed using the micro-fraction collection capabilities of the autosampler allowing automatic fractionation of intact proteins after the weak-ion-exchange (WAX) separation, and re-injection of the fractions onto the second-dimension RP monolithic column. The best peak capacity-to-analysis time ratio was obtained when applying 10 min second-dimension RP gradients. At optimized conditions, the WAX/x/RPLC separation of intact Escherichia coli proteins was performed within 6 h yielding a maximum theoretical peak capacity of 4880. PMID- 19557813 TI - Determination of aromatic amines from azo dyes reduction by liquid-phase sorbent trapping and thermal desorption-gas chromatography-mass spectrometry. AB - A novel method based on thermal desorption was developed for the trapping of aromatic amines from azo dyes reduction in liquid phase. Combining GC-MS, the detection limits for 21 kinds of aromatic amines ranged from 0.1 to 3 mg/kg. The recoveries of most aromatic amines were > or = 70% with RSDs < or = 10%. This method also offers low sample and organic solvent requirements, effective preconcentration and convenient procedure. It has been applied to determine aromatic amines from the real samples of textile and yielded good results. PMID- 19557814 TI - Chiral separation of amides of amino acid on a (S)-N-(3,5-dinitrobenzoyl)leucine N-phenyl-N-propylamide-bonded silica using nonaqueous capillary electrochromatography. AB - (S)-N-(3,5-dinitrobenzoyl)leucine-N-phenyl-N-propylamine-bonded silica was used as a chiral stationary phase for separation of a set of racemic pi-acidic and pi basic alpha-amino acid amides in electrolyteless ACN-water eluents by CEC in the RP and polar organic (PO) modes. The effect of the amount of water in the ACN water eluent on chiral separation was examined. As water is added to ACN, retention was shortened but resolution and selectivity deteriorated severely. Retention, enantioselectivity, and resolution factors obtained in 100% ACN were compared with those in an n-hexane-isopropanol eluent with a small amount of water by normal phase (NP) CEC. Much shorter retention times with comparable enantioselectivities were observed with 100% ACN, demonstrating the advantage of separation on (S)-N-(DNB)leucine-N-phenyl-N-propylamine-bonded silica in PO-CEC over NP-CEC. PMID- 19557815 TI - Microwave-assisted extraction with water for fast extraction and simultaneous RP HPLC determination of phenolic acids in radix Salviae Miltiorrhizae. AB - An optimized microwave-assisted extraction method using water (MAE-W) as the extractant and an efficient HPLC analysis method were first developed for the fast extraction and simultaneous determination of D(+)-(3,4-dihydroxyphenyl) lactic acid (Dla), salvianolic acid B (SaB), and lithospermic acid (La) in radix Salviae Miltiorrhizae. The key parameters of MAE-W were optimized. It was found that the degradation of SaB was inhibited when using the optimized MAE-W and the stable content of Dla, La, and SaB in danshen was obtained. Furthermore, compared to the conventional extraction methods, the proposed MAE-W is a more rapid method with higher yield and lower solvent consumption with a reproducibility (RSD <6%). In addition, using water as extractant is safe and helpful for environment protection, which could be referred to as green extraction. The separation and quantitative determination of the three compounds was carried out by a developed reverse-phase high-performance liquid chromatographic (RP-HPLC) method with UV detection. Highly efficient separation was obtained using gradient solvent system. The optimized HPLC analysis method was validated to have specificity, linearity, precision, and accuracy. The results indicated that MAE-W followed by HPLC-UV determination is an appropriate alternative to previously proposed method for quality control of radix Salviae Miltiorrhizae. PMID- 19557816 TI - Capillary electrophoresis of intact basic proteins using noncovalently triple layer coated capillaries. AB - The usefulness of a noncovalent, positively charged capillary coating for the efficient analysis of intact basic proteins with CE was studied. Capillaries were coated by subsequent flushing with solutions of 10% w/v Polybrene (PB), 3% w/v dextran sulfate (DS), and again 10% w/v PB. Coating characterization studies showed that stable coatings could be produced which exhibited a pH-independent and highly reproducible EOF. The PB-DS-PB coating was evaluated with Tris phosphate BGEs of various pH using the four basic model proteins: alpha chymotrypsinogen A, ribonuclease A, cytochrome c, and lysozyme. Typical migration time RSDs for the proteins were less than 0.85%, and apparent plate numbers were above 125,000 using a capillary length of 40 cm. The high separation efficiency allowed detection of several minor impurities in the model proteins. Using a BGE of medium pH, the CE system with triple-layer coating appeared to be useful for the repeatable profiling of recombinant humanized mouse monoclonal immunoglobulin G(1) showing a characteristic pattern of glycoforms. The CE system was also applied to the characterization of two llama antibodies, which were produced in Saccharomyces cerevisiae, revealing the presence of a side product in one of the antibodies. The high migration time stability allowed the reliable determination of antibody-antigen binding by monitoring migration time shifts. Finally, the feasibility of using the PB-DS-PB coated capillaries for CE with mass spectrometric detection was shown by the characterization of the impure llama antibody sample. PMID- 19557817 TI - Determination of penicillins in milk using LC-UV, LC-MS and LC-MS/MS. AB - The aim of this work is to establish a method for the simultaneous determination of eight penicillins in milk samples by LC-UV, LC-MS and LC-MS/MS. The procedure involves a step for clean-up and to preconcentrate the analytes by SPE and a subsequent chromatographic analysis. LC-UV, LC-MS and LC-MS/MS have been used for the simultaneous quantification of penicillins in milk. The proposed methods have been validated according to the EU guideline and present LOQ below the maximum limits of residues (MRLs) established by the European Union for penicillins in milk. The developed methods were applied to different milk samples obtained from cows medicated with penicillins. PMID- 19557818 TI - Aspects of food processing and its effect on allergen structure. AB - The article summarizes current physical and chemical methods in food processing as storage, preparation, separation, isolation or purification and thermal application on the one hand as well as enzymatic treatment on the other and their impact on the properties of food proteins. Novel methods of food processing like high pressure, electric field application or irradiation and their impact on food allergens are presented. The EU project REDALL (Reduced Allergenicity of Processed Foods, Containing Animal Allergens: QLK1-CT-2002-02687) showed that by a combination of enzyme and heat treatment the allergic potential of hen's egg decreased about 100 fold. Clinical reactions do not appear anymore. An AiF-FV 12024 N project worked with fruits like mango, lychee and apple. Processed mango and lychee had no change in allergenic potential during heating while e. g. canning. Apple almost lost its allergenic potential after pasteurization in juice production. PMID- 19557819 TI - Cytotoxicity of Maillard reaction products determined with a peptide spot library. AB - The reaction of reactive carbonyl compounds (RCCs) with lysine and arginine (Maillard reaction) is a common modification of proteins in thermally processed foods. In this study, the toxicity of Maillard reaction products (MRPs) formed from defined amino acids or dipeptides (bound to a cellulose membrane) with ribose, glycerinaldehyde or methylglyoxal was investigated. Murine RAW 264.7 macrophages were cultivated on the cellulose membrane and the effect of MRPs on cell viability was determined. The toxicity of MRPs was dependent on the RCC used and increased in the order of ribose < glycerinaldehyde < methylglyoxal. The dipeptides were more cytotoxic than the amino acids, with Lys-Lys MRPs being the most toxic of all tested MRPs. Cell numbers did not fall below the starting point, indicating that the MRPs rather inhibited proliferation than actually caused cell death. To develop an assay, in which whole membranes with multiple peptide spots could be tested simultaneously, we measured cell numbers on larger cellulose membranes using image analysis of the intracellularly formed formazan crystals. Although this method was technically feasable, it appears that uneven cell attachment on the membrane would require a way to detemine starting cell number by a non-destructive assay to yield more robust data. PMID- 19557820 TI - Polyphenol metabolites from colonic microbiota exert anti-inflammatory activity on different inflammation models. AB - The polyphenols in fruits and vegetables may be partly responsible for the health promoting effects attributed to fruit and vegetable intake. Although their properties have been relatively well studied, the activity of their metabolites, produced after ingestion, has been poorly investigated. Thus, the aim of this work was to study the potential anti-inflammatory effect of 18 polyphenol metabolites, derived from colon microbiota. They were screened by measuring prostaglandin E(2) (PGE(2)) production by CCD-18 colon fibroblast cells stimulated with IL-1beta. Metabolites that inhibited more than 50% PGE(2) production were hydrocaffeic (HCAF), dihydroxyphenyl acetic (dOHPA), and hydroferulic acid (HFER), that subsequently were tested with the writhing and paw pressure test in rodents where all three compounds showed an anti-inflammatory effect. The effect of HCAF administered orally (50 mg/kg) was also tested in the dextran sodium sulfate (DSS)-induced colitis model. Weight loss and fecal water content were more pronounced in DSS rats than in DSS-HCAF treated rats. HCAF treatment diminished the expression of the cytokines IL-1beta, IL-8, and TNF alpha, reduced malonyldialdehyde (MDA) levels and oxidative DNA damage (measured as 8-oxo-2'-deoxyguanosine levels) in distal colon mucosa. These results indicate that HCAF, dOHPA, and HFER have anti-inflammatory activity in vitro and in vivo. PMID- 19557821 TI - Naturally occurring flavonoids attenuate high glucose-induced expression of proinflammatory cytokines in human monocytic THP-1 cells. AB - Activation of circulating monocytes by hyperglycemia is bound to play a role in inflammatory and atherosclerosis. In this study, we examined whether flavonoids (catechin, EGCG, luteolin, quercetin, rutin) - phytochemicals that may possible belong to a new class of advanced glycation end products (AGEs) inhibitors - can attenuate high glucose (15 mmol/L, HG)-induced inflammation in human monocytes. Our results show that all flavonoids significantly inhibited HG-induced expression of proinflammatory genes and proteins, including TNF-alpha, interleukin-1beta (IL-1beta), and cyclooxygenase (COX)-2, at a concentration of 20 microM. Flavonoids also prevented oxidative stress in activated monocytes, as demonstrated by their inhibitory effects on intracellular reactive oxygen species (ROS) and N(epsilon)-(carboxymethyl)lysine formation caused by HG. These inhibitory effects may involve inhibition of nuclear factor-kappaB activation and may be supported by downregulation of the following: i) PKC-dependent NADPH oxidase pathway; ii) phosphorylation of p38 mitogen-activated protein kinase and extracellular signal-regulated protein kinase, and iii) mRNA expression of receptor of AGEs. In addition, we found for the first time that lower levels of Bcl-2 protein under HG conditions could be countered by the action of flavonoids. Our data suggest that, along with their antioxidant activities, flavonoids possess anti-inflammatory properties and might therefore have additional protective effects against glycotoxin-related inflammation. PMID- 19557822 TI - Inhibition of lung cancer growth in mice by dietary mixed tocopherols. AB - Tocopherols are lipophilic antioxidants found in vegetable oils. Here, we examined the growth inhibitory effect of a gamma-tocopherol-enriched tocopherol mixture (gammaTmT) against CL13 murine lung cancer cells grown in culture and as subcutaneous tumors in A/J mice. We found gammaTmT had no effect after 2 days and weakly inhibited the growth of CL13 in culture after 5 days (28% growth inhibition at 80 microM). Dietary treatment with 0.1 and 0.3% gammaTmT for 50 days inhibited the growth of CL13 tumors in A/J mice by 53.9 and 80.5%, respectively. Histopathological analysis revealed an increase in tumor necrosis compared to control tumors (80 and 240% increase by 0.1 and 0.3% gammaTmT, respectively). Dietary treatment with gammaTmT dose-dependently increased gamma- (10.0-37.6-fold) and delta-tocopherol (8.9-26.7-fold) in the tumors of treated mice compared to controls. Dietary treatment with gammaTmT also increased plasma gamma- (5.4-6.7-fold) and delta-tocopherol (5.5-7-fold). Whereas others have demonstrated the cancer preventive activity of gammaTmT against mammary and colon cancer, this is the first report of growth inhibitory activity against lung cancer. Further studies are needed to determine the underlying mechanisms for this anticancer activity, and to determine if such activity occurs in other models of cancer. PMID- 19557823 TI - Predictors of distress and quality of life in patients undergoing cancer therapy: impact of treatment type and decisional role. AB - PURPOSE: The purpose of this secondary investigation was to examine the impact of the type of treatment received and the perceived role in treatment decision making in predicting distress and cancer-specific quality of life in patients newly diagnosed with breast or prostate cancer. METHOD: Participants included 1057 newly diagnosed breast and prostate cancer patients from four Canadian cancer centers who partook in a randomized controlled trial examining the utility of providing patients with an audio-recording of their treatment planning consultation. A MANCOVA was performed to predict distress and cancer-specific quality of life at 12 weeks post-consultation based on control variables (patient age, education, residence, tumor size (breast sample), gleason score (prostate sample), and receipt of an initial treatment consultation recording), predictor variables (treatment type--chemotherapy, hormone therapy, radiation therapy; decisional role--active, collaborative, passive), and interactions between these predictors. RESULTS: Women who received chemotherapy and reported having played a more passive role in treatment decision making had significantly greater distress and lower cancer-specific quality of life at 12-week post-consultation. There were no statistically significant predictors of these outcomes identified for men with prostate cancer. CONCLUSION: Receipt of chemotherapy places women with breast cancer at risk for distress and reduced quality of life, but only for the subset of women who report playing a passive role in treatment decision making. Prospective, longitudinal studies are needed to confirm the present findings and to explicate the antecedents, composition, and consequences of the 'passive' decisional role during the treatment phase of the cancer trajectory. PMID- 19557824 TI - Are they just lazy? Student teachers' attitudes about dyslexia. AB - It is highly likely that teachers' abilities in dealing with different forms of learning difficulties will be affected by their knowledge about and attitudes towards those difficulties. Ajzen's Theory of Planned Behaviour (TPB) provides a useful framework within which to explore such attitudes and was used as the starting point for investigating trainee teachers' attitudes towards aspects of dyslexia. It is assumed that new teachers will enter the profession with a set of intentions according to personal beliefs, normative views within the teaching profession and the sense of competence and power they feel in dealing with dyslexic difficulties. The purpose of this study was to probe student teachers' attitudes at a prestigious School of Education in the Southwest of England. Four hundred and eight primary and secondary Post Graduate Certificate in Education (PGCE) students responded to a survey asking about their attitudes toward dyslexia. The student teachers expressed strongly positive attitudes toward the construct of dyslexia, with the majority expressing confidence in their ability to support dyslexic pupils. Females held significantly more positive attitudes toward dyslexia than males, but there were no significant differences according to PGCE course subject. Students who took the survey before and after teaching practice demonstrated some small but significant changes in attitude scores over that time. It is proposed that a new breed of teachers may be entering the teaching profession with positive beliefs about their ability to help dyslexic pupils, but who remain unclear as to how this can be accomplished. Some implications for action and suggestions for future research are provided. PMID- 19557825 TI - Personal control over the cure of breast cancer: adaptiveness, underlying beliefs and correlates. AB - OBJECTIVES: Although cognitive adaptation theory suggests that personal control acts as a stress buffer when facing adversity, maladaptive outcomes might occur when control is disconfirmed. The moderating effect of disappointing news on the adaptiveness of personal control over cure in women with breast cancer was examined and contrasted with the effect on the adaptiveness of general control over life. Additionally, the underlying beliefs and correlates of control over cure were explored. METHODS: Women with newly diagnosed breast cancer were assessed after surgery (n=228). For a sub-sample (n=133) data before surgery and after the end of treatment were available as well. Data were collected through questionnaires and face-to-face interviews. The prescription of chemotherapy after surgery was used as an indicator of disappointing news. RESULTS: A chemotherapy prescription neither enhances nor limits the adaptiveness of disease specific or general control perceptions. Women reported that maintaining a positive attitude, accepting treatment and adopting a healthy life style gave them a sense of control over cure. Women with a strong sense of control over cure more often had invasive cancer, were younger and were best characterized by high optimism and strong sense of control over life. CONCLUSIONS: The findings add to our understanding of exaggerated control perceptions in cancer patients treated with curative intent and do not give reason to assume that such perceptions should be altered because of potentially maladaptive effects. On the contrary, a strong sense of control over the cure of breast cancer seems to reflect the capacity to adapt. PMID- 19557826 TI - A potentially dynamic lysosomal role for the endogenous TRPML proteins. AB - Lysosomal storage disorders (LSDs) constitute a diverse group of inherited diseases that result from lysosomal storage of compounds occurring in direct consequence to deficiencies of proteins implicated in proper lysosomal function. Pathology in the LSD mucolipidosis type IV (MLIV), is characterized by lysosomal storage of lipids together with water-soluble materials in cells from every tissue and organ of affected patients. Mutations in the mucolipin 1 (TRPML1) protein cause MLIV and TRPML1 has also been shown to interact with two of its paralogous proteins, mucolipin 2 (TRPML2) and mucolipin 3 (TRPML3), in heterologous expression systems. Heterogeneous lysosomal storage is readily identified in electron micrographs of MLIV patient cells, suggesting that proper TRPML1 function is essential for the maintenance of lysosomal integrity. In order to investigate whether TRPML2 and TRPML3 also play a role in the maintenance of lysosomal integrity, we conducted gene-specific knockdown assays against these protein targets. Ultrastructural analysis revealed lysosomal inclusions in both TRPML2 and TRPML3 knockdown cells, suggestive of a common mechanism for these proteins, in parallel with TRPML1, in the regulation of lysosomal integrity. However, co-immunoprecipitation assays revealed that physical interactions between each of the endogenous TRPML proteins are quite limited. In addition, we found that all three endogenous proteins only partially co-localize with each other in lysosomal as well as extra-lysosomal compartments. This suggests that native TRPML2 and TRPML3 might participate with native TRPML1 in a dynamic form of lysosomal regulation. Given that depletion of TRPML2/3 led to lysosomal storage typical to an LSD, we propose that depletion of these proteins might also underlie novel LSD pathologies not described hitherto. PMID- 19557827 TI - Progranulin expression correlates with dense-core amyloid plaque burden in Alzheimer disease mouse models. AB - Amyloid-beta (Abeta) plaques are pathological hallmarks of Alzheimer disease (AD). In addition, innate inflammatory responses, such as those mediated by microglia, are integral to the pathogenesis of AD. Interestingly, only dense-core plaques and not diffuse plaques are associated with neuritic and inflammatory pathology in AD patients as well as in mouse AD models. However, the precise neuropathological changes that occur in the brain in response to amyloid deposition are largely unknown. To study the molecular mechanism(s) responsible for Abeta-mediated neuropathology, we performed a gene expression analysis on laser-microdissected brain tissue of Tg2576 and APPPS1 mice that are characterized by different types of amyloid plaques and genetic backgrounds. Data were validated by image and biochemical analyses on different ages of Tg2576, APPPS1, and Abeta42-depositing BRI-Abeta42 mice. Consistent with an important role of inflammatory responses in AD, we identified progranulin (mouse Grn; human GRN) as one of the top ten up-regulated molecules in Tg2576 ( approximately 8 fold increased) and APPPS1 ( approximately 2-fold increased) mice compared to littermate controls, and among the eight significantly up-regulated molecules common to both mouse models. In addition, Grn levels correlated significantly with amyloid load, especially the dense-core plaque pathology (p < 0.001). We further showed that Grn is up-regulated in microglia and neurons and neurites around dense-core plaques, but not in astrocytes or oligodendrocytes, as has been shown in AD patients. Our data therefore support the ongoing use of these mouse models in drug trials, especially those with anti-inflammatory compounds. Moreover, the correlation of Grn with increasing disease severity in AD mouse models prompts human studies exploring the viability of GRN as a disease biomarker. Because loss of GRN has recently been shown to cause frontotemporal dementia and serves as a risk factor for AD, the strong GRN reactivity around dense-core plaques is consistent with an important role of this factor in AD pathogenesis. PMID- 19557828 TI - Down-regulated expression of SATB2 is associated with metastasis and poor prognosis in colorectal cancer. AB - To identify novel biomarkers of metastasis of colorectal cancer (CRC), we developed an orthotopic implantation model of murine CRC and selected in vivo M5, a subclone of the SW480 CRC cell line with enhanced potential for metastasis to the liver. We compared the differences in the gene expression profiles between M5 and SW480 cells using gene expression profiling. We found that expression of special AT-rich sequence-binding protein 2 (SATB2) was down-regulated in M5 cells. Immunohistochemical analysis of 146 colorectal tumour samples showed that underexpression of SATB2 was strongly correlated with poor prognosis, tumour invasion, lymph node metastasis, distant metastasis, and Dukes' classification for CRC. Univariate and multivariate survival analyses further showed that SATB2 expression was a potential favourable prognostic factor for CRC. These results demonstrated not only that SATB2 is a potential novel prognostic factor for CRC, but also that selection of a highly metastatic clone of SW480 in vivo coupled with gene expression profiling is a powerful approach to identifying prognostic markers for CRC. PMID- 19557829 TI - Optimization of mass transfer for toxin removal and immunoprotection of hepatocytes in a bioartificial liver. AB - This study was designed to determine optimal operating conditions of a bioartificial liver (BAL) based on mass transfer of representative hepatotoxins and mediators of immune damage. A microprocessor-controlled BAL was used to study mass transfer between patient and cell compartments separated by a hollow fiber membrane. Membrane permeability (70, 150, or 400 kDa molecular weight cut-off MWCO), membrane convection (high: 50 mL/min; medium: 25 mL/min; low: 10 mL/min; diffusion: 0 mL/min), and albumin concentration in the cell compartment (0.5 or 5 g%) were considered for a total of 24 test conditions. Initially, the patient compartment contained pig plasma supplemented with ammonia (0.017 kDa), unconjugated bilirubin (0.585 kDa), conjugated bilirubin (0.760 kDa), TNF-alpha (17 kDa), pig albumin (67 kDa), pig IgG (147 kDa), and pig IgM (900 kDa). Mass transfer of each substance was determined by its rate of appearance in the cell compartment. Membrane fouling was assessed by dextran polymer technique. Of the three tested variables (membrane pore size, convection, and albumin concentration), membrane permeability had the greatest impact on mass transfer (P < 0.001). Mass transfer of all toxins was greatest under high convection with a 400 kDa membrane. Transfer of IgG and IgM was insignificant under all conditions. Bilirubin transfer was increased under high albumin conditions (P = 0.055). Fouling of membranes ranged from 7% (400 kDa), 24% (150 kDa) to 62% (70 kDa) during a 2-h test interval. In conclusion, optimal toxin removal was achieved under high convection with a 400-kDa membrane, a condition which should provide adequate immunoprotection of hepatocytes in the BAL. PMID- 19557830 TI - Plasma facilitated delivery of DNA to skin. AB - Non-viral delivery of cell-impermeant drugs and DNA in vivo has traditionally relied upon either chemical or physical stress applied directly to target tissues. Physical methods typically use contact between an applicator, or electrode, and the target tissue and may involve patient discomfort. To overcome contact-dependent limitations of such delivery methodologies, an atmospheric helium plasma source was developed to deposit plasma products onto localized treatment sites. Experiments performed in murine skin showed that samples injected with plasmid DNA encoding luciferase and treated with plasma demonstrated increased levels of expression relative to skin samples that received injections of DNA alone. Increased response relative to injection alone was observed when either positive or negative voltage was used to generate the helium plasma. Quantitative results over a 26-day follow-up period showed that luciferase levels as high as 19-fold greater than the levels obtained by DNA injection alone could be achieved. These findings indicate that plasmas may compete with other physical delivery methodologies when skin is the target tissue. PMID- 19557831 TI - Endothelial cells support survival, proliferation, and neuronal differentiation of transplanted adult ischemia-induced neural stem/progenitor cells after cerebral infarction. AB - Transplantation of neural stem cells (NSCs) has been proposed as a therapy for a range of neurological disorders. To realize the potential of this approach, it is essential to control survival, proliferation, migration, and differentiation of NSCs after transplantation. NSCs are regulated in vivo, at least in part, by their specialized microenvironment or "niche." In the adult central nervous system, neurogenic regions, such as the subventricular and subgranular zones, include NSCs residing in a vascular niche with endothelial cells. Although there is accumulating evidence that endothelial cells promote proliferation of NSCs in vitro, there is no description of their impact on transplanted NSCs. In this study, we grafted cortex-derived stroke-induced neural stem/progenitor cells, obtained from adult mice, onto poststroke cortex in the presence or absence of endothelial cells, and compared survival, proliferation, and neuronal differentiation of the neural precursors in vivo. Cotransplantation of endothelial cells and neural stem/progenitor cells increased survival and proliferation of ischemia-induced neural stem/progenitor cells and also accelerated neuronal differentiation compared with transplantation of neural precursors alone. These data indicate that reconstitution of elements in the vascular niche enhances transplantation of adult neural progenitor cells. PMID- 19557832 TI - Using microarray technology to select housekeeping genes in Chinese hamster ovary cells. AB - In the present study, we have identified species-specific housekeeping genes (HKGs) for Chinese Hamster Ovary (CHO) cells using data from microarray gene expression profiling. HKGs suitable for quantitative RT-PCR normalization should display relatively stable expression levels across experimental conditions. We analyzed transcription profiles of several IgG-producing recombinant CHO cell lines under numerous culture conditions using a custom CHO DNA microarray platform and calculated relative expression variability from 124 microarrays. We selected a novel panel of candidate HKGs based on their low variability in expression from the microarray data. Compared to three traditional HKGs (Gapdh, Actb, and B2m), the majority of genes on this panel demonstrated lower or equal variability. Each candidate HKG was then validated using qRT-PCR. Final selection of CHO-specific HKGs include Actr5, Eif3i, Hirip3, Pabpn1, Vezt, Cog1, and Yaf2. The results reported here provide a useful tool for gene expression analyses in CHO cells, a critical expression platform used in biotherapeutics. PMID- 19557833 TI - The generation of stable, high MAb expressing CHO cell lines based on the artificial chromosome expression (ACE) technology. AB - The manufacture of recombinant proteins at industrially relevant levels requires technologies that can engineer stable, high expressing cell lines rapidly, reproducibly and with relative ease. Commonly used methods incorporate transfection of mammalian cell lines with plasmid DNA containing the gene of interest. Identifying stable high expressing transfectants is normally laborious and time consuming. To improve this process, the ACE System has been developed based on pre-engineered artificial chromosomes with multiple recombination acceptor sites. This system allows for the targeted transfection of single or multiple genes and eliminates the need for random integration into native host chromosomes. To illustrate the utility of the ACE System in generating stable, high expressing cell lines, CHO based candidate cell lines were generated to express a human monoclonal IgG1 antibody. Candidate cell lines were generated in under 6 months and expressed over 1 g/L and with specific productivities of up to 45 pg/cell/day under non-fed, non-optimized shake flask conditions. These candidate cell lines were shown to have stable expression of the monoclonal antibody for up to 70 days of continuous culture. The results of this study demonstrate that clonal, stable monoclonal antibody expressing CHO based cell lines can be generated by the ACE System rapidly and perform competitively with those cell lines generated by existing technologies. The ACE System, therefore, provides an attractive and practical alternative to conventional methods of cell line generation. PMID- 19557834 TI - The role of feticide in the context of late termination of pregnancy: a qualitative study of health professionals' and parents' views. AB - OBJECTIVE: To provide an in-depth account of the role feticide has relative to experiences of late termination of pregnancy (TOP). METHOD: Exploratory qualitative interview study. Participants were recruited from three National Health Service (NHS) units that provide secondary and tertiary level fetal medicine services. Data were collected from 36 in-depth interviews, with 12 parents (representing eight cases) who had experienced late TOP for fetal anomaly and 23 health professionals with experience of feticide provision. The qualitative analysis utilised a generative thematic approach, facilitated by Atlas.ti qualitative software package. RESULTS: Two key themes from the study provide data on how perceptions of feticide were described by those involved in late TOP: (1) feticide is recognised and described as a legitimate clinical procedure and (2) the practice of feticide is conceptualised as difficult but necessary. CONCLUSION: For health professionals who provide and facilitate feticide, and for parents making decisions about late TOP and feticide, the procedure is understood as a necessary rather than chosen activity. Parents' perceptions of feticide may differ, and good clinical care must be designed to cope with this variation. For health professionals, feticide seems more readily distinguished from other types of TOP activities and may evoke simultaneous positive and negative perceptions. PMID- 19557835 TI - Cell-free synthesis of functional aquaporin Z in synthetic liposomes. AB - The challenges involved in producing sufficient quantities of aquaporins for precise biophysical characterization have limited our knowledge of this important class of molecules. This article describes a cell-free protein synthesis method for producing high concentrations of the E. coli water transporter, aquaporin Z (AqpZ), in synthetic liposomes. To our knowledge, this is the first report of in vitro synthesis of a membrane protein directly into synthetic liposomes with verified function, (i.e., transport activity and selectivity). Titration of DOPC lipid vesicles added to the cell-free reaction show that production yields of active AqpZ are dependent on the concentration of DOPC lipid vesicles added to the cell-free reaction, with 224 +/- 24 lipids required per aquaporin monomer. Supplementation of the signal recognition particle receptor (FtsY) to the cell free reaction increases production of vesicle-associated AqpZ but not active AqpZ. Cell-free reactions using 7 mg/mL lipids that were not supplemented with FtsY produced 507 +/- 11 microg/mL of vesicle-associated AqpZ that exhibited a specific water transport activity of (2.2 +/- 0.3) x 10(-14) cm(3) s(-1) monomer( 1). Proteinase K protection, activation energy determination, and selectivity against glycerol and urea transport also confirmed the production of correctly folded AqpZ. This technique is capable of producing milligram quantities of aquaporin that can be readily assayed for function, facilitating biophysical characterization and high-throughput analysis. PMID- 19557836 TI - In vitro and in vivo RNA interference mediated suppression of Tn-caspase-1 for improved recombinant protein production in High Five cell culture with the baculovirus expression vector system. AB - While traditional metabolic engineering generally relies on the augmentation of specific genes and pathways in order to increase the yield of target proteins, the advent of RNA interference (RNAi) as a biological tool has given metabolic engineers another tool capable of rationally altering the host cell's biological landscape in order to achieve a specific goal. Given its broad applicability and potent specificity, RNAi has the ability to suppress genes whose function is contrary to the desired phenotype. In this study, RNAi has been used to increase recombinant protein production in a Trichoplusia ni derived cell line (BTI-TN-5B1 4-High Five) using the Baculovirus Expression Vector System. The specific target investigated is Tn-caspase-1, a protease involved in apoptosis that is likely the principal effector caspase present in T. ni cells. Experiments were first conducted using in vitro synthesized dsRNA to verify silencing of Tn-capase-1 and increased protein production as a result. Subsequent experiments were conducted using a cell line stably expressing in vivo RNAi in the form of an inverted repeat that results in a hairpin upon transcription. Using this construct, Tn caspase-1 transcript levels were decreased by 50% and caspase enzymatic activity was decreased by 90%. This cell line, designated dsTncasp-2, demonstrates superior viability under low nutrient culture conditions and resulted in as much as two times the protein yield when compared to standard High Five cells. PMID- 19557837 TI - Plasmid DNA production combining antibiotic-free selection, inducible high yield fermentation, and novel autolytic purification. AB - DNA vaccines and gene medicines, derived from bacterial plasmids, are emerging as an important new class of pharmaceuticals. However, the challenges of performing cell lysis processes for plasmid DNA purification at an industrial scale are well known. To address downstream purification challenges, we have developed autolytic Escherichia coli host strains that express endolysin (phage lambdaR) in the cytoplasm. Expression of the endolysin is induced during fermentation by a heat inducible promoter. The endolysin remains in the cytoplasm, where it is separated from its peptidoglycan substrate in the cell wall; hence the cells remain alive and intact and can be harvested by the usual methods. The plasmid DNA is then recovered by autolytic extraction under slightly acidic, low salt buffer conditions and treatment with a low concentration of non-ionic detergent. Under these conditions the E. coli genomic DNA remains associated with the insoluble cell debris and is removed by a solid-liquid separation. Here, we report fermentation, lysis methods, and plasmid purification using autolytic hosts. PMID- 19557838 TI - Bioimaging of metals by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). AB - The distribution analysis of (essential, beneficial, or toxic) metals (e.g., Cu, Fe, Zn, Pb, and others), metalloids, and non-metals in biological tissues is of key interest in life science. Over the past few years, the development and application of several imaging mass spectrometric techniques has been rapidly growing in biology and medicine. Especially, in brain research metalloproteins are in the focus of targeted therapy approaches of neurodegenerative diseases such as Alzheimer's and Parkinson's disease, or stroke, or tumor growth. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) using double focusing sector field (LA-ICP-SFMS) or quadrupole-based mass spectrometers (LA ICP-QMS) has been successfully applied as a powerful imaging (mapping) technique to produce quantitative images of detailed regionally specific element distributions in thin tissue sections of human or rodent brain. Imaging LA-ICP QMS was also applied to investigate metal distributions in plant and animal sections to study, for example, the uptake and transport of nutrient and toxic elements or environmental contamination. The combination of imaging LA-ICP-MS of metals with proteomic studies using biomolecular mass spectrometry identifies metal-containing proteins and also phosphoproteins. Metal-containing proteins were imaged in a two-dimensional gel after electrophoretic separation of proteins (SDS or Blue Native PAGE). Recent progress in LA-ICP-MS imaging as a stand-alone technique and in combination with MALDI/ESI-MS for selected life science applications is summarized. PMID- 19557839 TI - Metabolomic applications of HILIC-LC-MS. AB - Hydrophilic interaction liquid chromatography (HILIC), although not a new technique, has enjoyed a recent renaissance with the introduction of robust and reproducible stationary phases. It is consequently finding application in metabolomics studies, which have traditionally relied on the stability of reversed phases (RPs), since the biofluids analyzed are predominantly aqueous and thus contain many polar analytes. HILIC's retention of those polar compounds and use of solvents readily compatible with mass spectrometry have seen its increasing adoption in studies of complex aqueous metabolomes. This review describes the stationary phases and their features, surveys HILIC-LC-MS's role in metabolomics experiments, discusses approaches to data extraction and analysis including multivariate analysis, and reviews the literature on HILIC-MS applications in metabolomics. PMID- 19557840 TI - Automatic glioma characterization from dynamic susceptibility contrast imaging: brain tumor segmentation using knowledge-based fuzzy clustering. AB - PURPOSE: To assess whether glioma volumes from knowledge-based fuzzy c-means (FCM) clustering of multiple MR image classes can provide similar diagnostic efficacy values as manually defined tumor volumes when characterizing gliomas from dynamic susceptibility contrast (DSC) imaging. MATERIALS AND METHODS: Fifty patients with newly diagnosed gliomas were imaged using DSC MR imaging at 1.5 Tesla. To compare our results with manual tumor definitions, glioma volumes were also defined independently by four neuroradiologists. Using a histogram analysis method, diagnostic efficacy values for glioma grade and expected patient survival were assessed. RESULTS: The areas under the receiver operator characteristics curves were similar when using manual and automated tumor volumes to grade gliomas (P = 0.576-0.970). When identifying a high-risk patient group (expected survival <2 years) and a low-risk patient group (expected survival >2 years), a higher log-rank value from Kaplan-Meier survival analysis was observed when using automatic tumor volumes (14.403; P < 0.001) compared with the manual volumes (10.650-12.761; P = 0.001-0.002). CONCLUSION: Our results suggest that knowledge based FCM clustering of multiple MR image classes provides a completely automatic, user-independent approach to selecting the target region for presurgical glioma characterization. PMID- 19557841 TI - Impact of cerebrospinal fluid contamination on brain metabolites evaluation with 1H-MR spectroscopy: a single voxel study of the cerebellar vermis in patients with degenerative ataxias. AB - PURPOSE: To investigate the impact of cerebrospinal fluid (CSF) contamination on metabolite evaluation in the superior cerebellar vermis with single-voxel (1)H MRS in normal subjects and patients with degenerative ataxias. MATERIALS AND METHODS: Twenty-nine healthy volunteers and 38 patients with degenerative ataxias and cerebellar atrophy were examined on a 1.5 Tesla scanner. Proton spectra of a volume of interest placed in the superior vermis were acquired using a four TE PRESS technique. We calculated N-acetyl aspartate (NAA)/creatine (Cr), choline (Cho)/Cr, and NAA/Cho ratios, T(2) relaxation times and concentrations of the same metabolites using the external phantom method. Finally, concentrations were corrected taking into account the proportion of nervous tissue and CSF, that was determined as Volume Fraction (VF). RESULTS: In healthy subjects, a significant difference was observed between metabolite concentrations with and without correction for VF. As compared to controls, patients with ataxias showed significantly reduced NAA/Cr and NAA concentrations, while only corrected Cr concentration was significantly increased. The latter showed an inverse correlation with VF. CONCLUSION: CSF contamination has a not negligible effect on the estimation of brain metabolites. The increase of Cr concentration in patients with cerebellar atrophy presumably reflects the substitutive gliosis which takes place along with loss of neurons. PMID- 19557842 TI - Inhomogeneous sodium accumulation in the ischemic core in rat focal cerebral ischemia by 23Na MRI. AB - PURPOSE: To test the hypotheses that (i) the regional heterogeneity of brain sodium concentration ([Na(+)](br)) provides a parameter for ischemic progression not available from apparent diffusion coefficient (ADC) data, and (ii) [Na(+)](br) increases more in ischemic cortex than in the caudate putamen (CP) with its lesser collateral circulation after middle cerebral artery occlusion in the rat. MATERIALS AND METHODS: (23)Na twisted projection MRI was performed at 3 Tesla. [Na(+)](br) was independently determined by flame photometry. The ischemic core was localized by ADC, by microtubule-associated protein-2 immunohistochemistry, and by changes in surface reflectivity. RESULTS: Within the ischemic core, the ADC ratio relative to the contralateral tissue was homogeneous (0.63 +/- 0.07), whereas the rate of [Na(+)](br) increase (slope) was heterogeneous (P < 0.005): 22 +/- 4%/h in the sites of maximum slope versus 14 +/ 1%/h elsewhere (here 100% is [Na(+)](br) in the contralateral brain). Maximum slopes in the cortex were higher than in CP (P < 0.05). In the ischemic regions, there was no slope/ADC correlation between animals and within the same brain (P > 0.1). Maximum slope was located at the periphery of ischemic core in 8/10 animals. CONCLUSION: Unlike ADC, (23)Na MRI detected within-core ischemic lesion heterogeneity. PMID- 19557843 TI - A diffusion tensor imaging group study of the spinal cord in multiple sclerosis patients with and without T2 spinal cord lesions. AB - PURPOSE: To examine the T(2)-normal appearing spinal cord of patients with multiple sclerosis (MS) using diffusion tensor imaging. MATERIALS AND METHODS: Diffusion tensor images of the spinal cord were acquired from 21 healthy subjects, 11 MS patients with spinal cord lesions, and 10 MS patients without spinal cord lesions on the T(2)-weighted MR images. Different diffusion measures were evaluated using both a region of interest (ROI) -based and a diffusion tensor tractography-based segmentation approach. RESULTS: It was observed that the FA, the transverse diffusivity lambda(perpendicular), and the ratio of the longitudinal and transverse diffusivities (lambda(parallel)/lambda (perpendicular)) were significantly lower in the spinal cord of MS patients with spinal cord lesions compared with the control subjects using both the ROI method (P = 0.014, P = 0.028, and P = 0.039, respectively) and the tractography-based approach (P = 0.006, P = 0.037, and P = 0.012, respectively). For both image analysis methods, the FA and the lambda (parallel)/lambda (perpendicular) values were significantly different between the control group and the MS patient group without T(2) spinal cord lesions (P = 0.013). CONCLUSION: Our results suggest that the spinal cord may still be affected by MS, even when lesions are not detected on a conventional MR scan. In addition, we demonstrated that diffusion tensor tractography is a robust tool to analyze the spinal cord of MS patients. PMID- 19557844 TI - Contrast-enhanced MRI of carotid atherosclerosis: dependence on contrast agent. AB - PURPOSE: To investigate the dependence of contrast-enhanced magnetic resonance imaging (MRI) of carotid artery atherosclerotic plaque on the use of gadobenate dimeglumine versus gadodiamide. MATERIALS AND METHODS: Fifteen subjects with carotid atherosclerotic plaque were imaged with 0.1 mmol/kg of each agent. For arteries with interpretable images, the areas of the lumen, wall, and necrotic core and overlying fibrous cap (when present) were measured, as were the percent enhancement and contrast-to-noise ratio (CNR). A kinetic model was applied to dynamic imaging results to determine the fractional plasma volume, v(p), and contrast agent transfer constant, K(trans). RESULTS: For 12 subjects with interpretable images, the agent used did not significantly impact any area measurements or the presence or absence of necrotic core (P > 0.1 for all). However, the percent enhancement was greater for the fibrous cap (72% vs. 54%; P < 0.05) necrotic core (51% vs. 42%; P = 0.12), and lumen (42% vs. 63%; P < 0.05) when using gadobenate dimeglumine, although no apparent difference in CNR was found. Additionally, K(trans) was lower when using gadobenate dimeglumine (0.0846 min(-1) vs. 0.101 min(-1); P < 0.01), although v(p) showed no difference (9.5% vs. 10.1%; P = 0.39). CONCLUSION: Plaque morphology measurements are similar with either contrast agent, but quantitative enhancement characteristics, such as percent enhancement and K(trans), differ. PMID- 19557845 TI - Combining spatial and temporal information to explore function-guide action of acupuncture using fMRI. AB - PURPOSE: To investigate the brain response patterns of modulation of GB37 (Guangming) and KI8 (Jiaoxin). MATERIALS AND METHODS: An experiment using nonrepeated event-related fMRI design was carried out on 28 subjects with electroacupuncture stimulation (EAS) at GB37 or KI8 on the left leg. The discrete cosine transform and functional connectivity methods were adopted to detect the differences related with these two acupoints before and after the EAS. RESULTS: Spatial patterns were distinct for EAS at the two acupoints, and the overlapping brain regions were mainly located in the posterior cingulate cortex (PCC) and precuneus (pC). Two opposite patterns of modulation in the default mode network were detected from the temporal patterns with the overlapping PCC/pC as the region of interest. Furthermore, the specific responses of sustained effects at these acupoints were also identified. CONCLUSION: Spatial and temporal patterns of the sustained effect modulation of GB37 and KI8 were distinct. We suggest these findings may attribute to the functional specificity of a certain acupoint. Moreover, our current results reflect a significant methodological contribution to future acupuncture studies. PMID- 19557846 TI - Improving computation of cardiovascular relative pressure fields from velocity MRI. AB - PURPOSE: To evaluate a multigrid-based solver for the pressure Poisson equation (PPE) with Galerkin coarsening, which works directly on the specified domain, for the computation of relative pressure fields from velocity MRI data. MATERIALS AND METHODS: We compared the proposed structure-defined Poisson solver to other popular Poisson solvers working on unmodified rectangular and modified quasirectangular domains using synthetic and in vitro phantoms in which the mathematical solution of the pressure field is known, as well as on in vivo MRI velocity measurements of aortic blood flow dynamics. RESULTS: All three PPE solvers gave accurate results for convex computational domains. Using a rectangular or quasirectangular domain on a more complicated domain, like a c shape, revealed a systematic underestimation of the pressure amplitudes, while the proposed PPE solver, working directly on the specified domain, provided accurate estimates of the relative pressure fields. CONCLUSION: Popular iterative approaches with quasirectangular computational domains can lead to significant systematic underestimation of the pressure amplitude. We suggest using a multigrid-based PPE solver with Galerkin coarsening, which works directly on the structure-defined computational domain. This solver provides accurate estimates of the relative pressure fields for both simple and complex geometries with additional significant improvements with respect to execution speed. PMID- 19557847 TI - Multicenter validation of the magnetic resonance T2* technique for segmental and global quantification of myocardial iron. AB - PURPOSE: To assess the transferability of the magnetic resonance imaging (MRI) multislice multiecho T2(*) technique for global and segmental measurement of iron overload in thalassemia patients. MATERIALS AND METHODS: Multiecho T2(*) sequences were installed on six MRI scanners. Five healthy subjects (n = 30) were scanned at each site; five thalassemia major (TM) patients were scanned at the reference site and were rescanned locally (n = 25) within 1 month. T2(*) images were analyzed using previously validated software. RESULTS: T2(*) values of healthy subjects showed intersite homogeneity. On TM patients, for global heart T2(*) values the correlation coefficient was 0.97, coefficients of variation (CoV(s)) ranged from 0.04-0.12, and intraclass coefficients (ICC(s)) ranged from 0.94-0.99. The mean CoV and ICC for segmental T2(*) distribution were 0.198 and 88, respectively. CONCLUSION: The multislice multiecho T2(*) technique is transferable among scanners with good reproducibility. PMID- 19557848 TI - Persistent decline in longitudinal and radial strain after coronary microembolization detected on velocity encoded phase contrast magnetic resonance imaging. AB - PURPOSE: To use velocity-encoded phase contrast (PC) MRI in assessing the effect of coronary microembolization on longitudinal and radial myocardial strain. MATERIALS AND METHODS: A combined X-ray and MR system (XMR) was used for selective left anterior descending artery catheterization and microinfarct assessment in swine (n = 6). The embolized area at risk was defined on perfusion MRI followed by administration of a 7500 count (size = 100-300 microm) of the embolic agent. Quantification of strain and microinfarction was performed at 1 h and 1 week using PC-MRI and delayed enhancement (DE) MRI, respectively. At postmortem, sliced hearts were stained to define microinfarction. RESULTS: Baseline longitudinal and radial strain did not differ between area-at-risk and remote myocardium. The embolized territory (area at risk) showed significant decline in longitudinal strain from -11.5 +/- 3.2% to 1.8 +/- 2.5% at 1 h (P < 0.05) and -3.9 +/- 1.1% at 1 week (P < 0.05). Similarly, regional radial strain progressively declined from 23.6 +/- 2.5% at baseline to 12.5 +/- 3.7% at 1 h (P < 0.05) and 4.8 +/- 5.0% at 1 week (P < 0.01). The size of microinfarction was not significantly different between DE-MRI and histochemical staining. CONCLUSION: PC-MRI is sensitive in assessing changes in regional longitudinal and radial strain after coronary embolization. Longitudinal and radial strain of the hyperenhanced patchy microinfarction demonstrates persistent decline over the course of 1 week. PMID- 19557849 TI - Three-dimensional analysis of segmental wall shear stress in the aorta by flow sensitive four-dimensional-MRI. AB - PURPOSE: To assess the distribution and regional differences of flow and vessel wall parameters such as wall shear stress (WSS) and oscillatory shear index (OSI) in the entire thoracic aorta. MATERIALS AND METHODS: Thirty-one healthy volunteers (mean age = 23.7 +/- 3.3 years) were examined by flow-sensitive four dimensional (4D)-MRI at 3T. For eight retrospectively positioned 2D analysis planes distributed along the thoracic aorta, flow parameters and vectorial WSS and OSI were assessed in 12 segments along the vascular circumference. RESULTS: Mean absolute time-averaged WSS ranged between 0.25 +/- 0.04 N/m(2) and 0.33 +/- 0.07 N/m(2) and incorporated a substantial circumferential component (-0.05 +/- 0.04 to 0.07 +/- 0.02 N/m(2)). For each analysis plane, a segment with lowest absolute WSS and highest OSI was identified which differed significantly from mean values within the plane (P < 0.05). The distribution of atherogenic low WSS and high OSI closely resembled typical locations of atherosclerotic lesions at the inner aortic curvature and supraaortic branches. CONCLUSION: The normal distribution of vectorial WSS and OSI in the entire thoracic aorta derived from flow-sensitive 4D-MRI data provides a reference constituting an important perquisite for the examination of patients with aortic disease. Marked regional differences in absolute WSS and OSI may help explaining why atherosclerotic lesions predominantly develop and progress at specific locations in the aorta. PMID- 19557850 TI - Study of reproducibility of human arterial plaque reconstruction and its effects on stress analysis based on multispectral in vivo magnetic resonance imaging. AB - PURPOSE: To quantify the uncertainties of carotid plaque morphology reconstruction based on patient-specific multispectral in vivo magnetic resonance imaging (MRI) and their impacts on the plaque stress analysis. MATERIALS AND METHODS: In this study, three independent investigators were invited to reconstruct the carotid bifurcation with plaque based on MR images from two subjects to study the geometry reconstruction reproducibility. Finite element stress analyses were performed on the carotid bifurcations, as well as the models with artificially modified plaque geometries to mimic the image segmentation uncertainties, to study the impacts of the uncertainties to the stress prediction. RESULTS: Plaque reconstruction reproducibility was generally high in the study. The uncertainties among interobservers are around one or the subpixel level. It also shows that the predicted stress is relatively less sensitive to the arterial wall segmentation uncertainties, and more affected by the accuracy of lipid region definition. For a model with lipid core region artificially increased by adding one pixel on the lipid region boundary, it will significantly increase the maximum Von Mises Stress in fibrous cap (>100%) compared with the baseline model for all subjects. CONCLUSION: The current in vivo MRI in the carotid plaque could provide useful and reliable information for plaque morphology. The accuracy of stress analysis based on plaque geometry is subject to MRI quality. The improved resolution/quality in plaque imaging with newly developed MRI protocols would generate more realistic stress predictions. PMID- 19557852 TI - Quantitative contrast-enhanced perfusion measurements of the human lung using the prebolus approach. AB - PURPOSE: To investigate dynamic contrast-enhanced MRI (DCE-MRI) for quantification of pulmonary blood flow (PBF) and blood volume (PBV) using the prebolus approach and to compare the results to the global lung perfusion (GLP). MATERIALS AND METHODS: Eleven volunteers were examined by applying different contrast agent doses (0.5, 1.0, 2.0, and 3.0 mL gadolinium diethylene triamine pentaacetic acid [Gd-DTPA]), using a saturation-recovery (SR) true fast imaging with steady precession (TrueFISP) sequence. PBF and PBV were determined for single bolus and prebolus. Region of interest (ROI) evaluation was performed and parameter maps were calculated. Additionally, cardiac output (CO) and lung volume were determined and GLP was calculated as a contrast agent-independent reference value. RESULTS: The prebolus results showed good agreement with low-dose single bolus and GLP: PBF (mean +/- SD in units of mL/minute/100 mL) = single bolus 190 +/- 73 (0.5-mL dose) and 193 +/- 63 (1.0-mL dose); prebolus 192 +/- 70 (1.0-2.0 mL dose) and 165 +/- 52 (1.0-3.0-mL dose); GLP (mL/minute/100 mL) = 187 +/- 34. Higher single-bolus resulted in overestimated values due to arterial input function (AIF) saturation. CONCLUSION: The prebolus approach enables independent determination of appropriate doses for AIF and tissue signal. Using this technique, the signal-to-noise ratio (SNR) from lung parenchyma can be increased, resulting in improved PBF and PBV quantification, which is especially useful for the generation of parameter maps. PMID- 19557853 TI - Hemolymph ecdysteroids during the last three molt cycles of the blue crab, Callinectes sapidus: quantitative and qualitative analyses and regulation. AB - The profiles of circulating ecdysteroids during the three molt cycles prior to adulthood were monitored from the juvenile blue crab, Callinectes sapidus. Ecdysteroid patterns are remarkably similar in terms of peak concentrations ranging between 210-330 ng/ml hemolymph. Analysis of hemolymph at late premolt stage revealed six different types of ecdysteroids with ponasterone A (PoA) and 20-OH ecdysone (20-OH E) as the major forms. This ecdysteroid profile was consistent in all three molt cycles. Bilateral eyestalk ablation (EA) is a procedure that removes inhibitory neurohormones including crustacean hyperglycemic hormone (CHH) and molt-inhibiting hormone (MIH) and often results in precocious molting in crustaceans. However, the inhibitory roles of these neuropeptides in vivo have not yet been tested in C. sapidus. We determined the regulatory roles of CHH and MIH in the circulating ecdysteroid from ablated animals through daily injection. A daily administration of purified native CHH and MIH at physiological concentration maintained intermolt levels of ecdysteroids in the EA animals. This suggests that Y organs (YO) require a brief exposure to CHH and MIH in order to maintain the low level of ecdysteroids. Compared to intact animals, the EA crabs did not exhibit the level of peak ecdysteroids, and the major ecdysteroid turned out to be 20-OH E, not PoA. These results further underscore the important actions of MIH and CHH in ecdysteroidogenesis, as they not only inhibit, but also control the composition of output of the YO activity. PMID- 19557851 TI - Cine cardiac imaging using black-blood steady-state free precession (BB-SSFP) at 3T. AB - PURPOSE: To propose a new black-blood (BB) pulse sequence that provides BB cine cardiac images with high blood-myocardium contrast. The proposed technique is based on the conventional steady-state free precession (SSFP) sequence. MATERIALS AND METHODS: Numerical simulations of the Bloch equation were conducted to compare the resulting signal-to-noise ratio (SNR) to that of conventional BB imaging, including the effects of changing the imaging flip angle and heart rates. Simulation results were verified using a gel phantom experiment and five normal volunteers were scanned using the proposed technique. RESULTS: The new sequence showed higher SNR and contrast-to-noise ratio (CNR) (approximately 100%) compared to the conventional BB imaging. Also, the borders of the left ventricle (LV) and right ventricle (RV) appear more distinguishable than the conventional SSFP. We were also able to cover about 80% of the cardiac cycle with short breath hold time (approximately 10 cardiac cycles) and with reasonable SNR and CNR. CONCLUSION: Based on an SSFP conventional sequence, the new sequence provides BB cines that cover most of the cardiac cycle and with higher SNR and CNR than the conventional BB sequences. PMID- 19557854 TI - Juvenile hormone-stimulated synthesis of acyl-glycerols and vitamin E in female accessory sexual glands of the fire bug, Pyrrhocoris apterus L. AB - Secretory cells of the female accessory sexual glands (AG) of P. apterus grow and produce yellow oily exocrine secretion in response to stimulation by endogenous juvenile hormone (JH) or exogenous treatments by JH analogues. The secretion determines the property of future egg shells by coating the chorion surface of the oocytes that are passing individually through the common uterus during oviposition. Diapausing females with a physiologically inhibited endocrine system or females with artificially removed hormonal sources show inactive ovaries and empty AG without the secretory products. Ovary-ectomised females with the intact neuroendocrine system develop hypertrophic AG loaded with the oily secretion. This shows that there is no direct dependence between formation of the oily secretion in AG and ovarian growth. Chemical analysis of the secretory products revealed the presence of acetylated glycerols, with the most abundant stearoyl diacetyl-glycerol, stearoyl-acetyl-propionyl-glycerol, and the corresponding derivatives of arachidonic acid. In addition to this, the JH-activated secretory cells of AG also produced gamma- and delta-tocopherols. The possible antioxidant or antimutagenic action of these vitamin E compounds in insect reproduction has been emphasized. PMID- 19557855 TI - Novel quinolone CHM-1 induces apoptosis and inhibits metastasis in a human osterogenic sarcoma cell line. AB - Novel 2-phenyl-4-quinolone compounds have potent cytotoxic effects on different human cancer cell lines. In this study, we examined anticancer activity and mechanisms of 20-fluoro-6,7-methylenedioxy-2-phenyl-4-quinolone (CHM-1) in human osterogenic sarcoma U-2 OS cells. CHM-1-induced apoptosis was determined by flow cytometric analysis, DAPI staining, Comet assay, and caspase inhibitors. CHM-1 inhibited cell migration and invasion was assessed by a wound healing assay, gelatin zymography, and a Transwell assay. The mechanisms of CHM-1 effects on apoptosis and metastasis signaling pathways were studied using Western blotting and gene expression. CHM-1 induced G2/M arrest and apoptosis at an IC(50) (3 microM) in U-2 OS cells and caspase-3, -8, and -9 were activated. Caspase inhibitors increased cell viability after exposure to CHM-1. CHM-1-induced apoptosis was associated with enhanced ROS generation, DNA damage, decreased DeltaPsi(m) levels, and promotion of mitochondrial cytochrome c release. CHM-1 stimulated mRNA expression of caspase-3, -8, and -9, AIF, and Endo G. In addition, CHM-1 inhibited cell metastasis at a low concentration (<3 microM). CHM 1 inhibited the cell metastasis through the inhibition of MMP-2, -7, and -9. CHM 1 also decreased the levels of MAPK signaling pathways before leading to the inhibition of MMPs. In summary, CHM-1 is a potent inducer of apoptosis, which plays a role in the anticancer activity of CHM-1. PMID- 19557856 TI - Free sialic acid storage disease without sialuria. AB - We performed high-resolution in vitro proton nuclear magnetic resonance spectroscopy on cerebrospinal fluid and urine samples of 44 patients with leukodystrophies of unknown cause. Free sialic acid concentration was increased in cerebrospinal fluid of two siblings with mental retardation and mild hypomyelination. By contrast, urinary excretion of free sialic acid in urine was normal on repeated testing by two independent methods. Both patients were homozygous for the K136E mutation in SLC17A5, the gene responsible for the free sialic acid storage diseases. Our findings demonstrate that mutations in the SLC17A5 gene have to be considered in patients with hypomyelination, even in the absence of sialuria. PMID- 19557857 TI - De novo STXBP1 mutations in mental retardation and nonsyndromic epilepsy. AB - We sequenced genes coding for components of the SNARE complex (STX1A, VAMP2, SNAP25) and their regulatory proteins (STXBP1/Munc18-1, SYT1), which are essential for neurotransmission, in 95 patients with idiopathic mental retardation. We identified de novo mutations in STXBP1 (nonsense, p.R388X; splicing, c.169+1G>A) in two patients with severe mental retardation and nonsyndromic epilepsy. Reverse transcriptase polymerase chain reaction and sequencing showed that the splicing mutation creates a stop codon downstream of exon-3. No de novo or deleterious mutations in STXBP1 were found in 190 control subjects, or in 142 autistic patients. These results suggest that STXBP1 disruption is associated with autosomal dominant mental retardation and nonsyndromic epilepsy. PMID- 19557858 TI - Seeing the phantom: a functional magnetic resonance imaging study of a supernumerary phantom limb. AB - OBJECTIVE: Supernumerary phantom limb (SPL) is a rare neurological manifestation where patients with a severe stroke-induced sensorimotor deficit experience the illusory presence of an extra limb that duplicates a real one. The illusion is most often experienced as a somesthetic phantom, but rarer SPLs may be intentionally triggered or seen. Here, we report the case of a left visual, tactile, and intentional SPL caused by right subcortical damage in a nondeluded woman. METHODS: Using functional magnetic resonance imaging, we investigated the multimodal nature of this phantom, which the patient claimed to be able see, use, and move intentionally. The patient participated in a series of sensorimotor and motor imagery tasks involving the right, the left plegic, and the SPL's hand. RESULTS: Right premotor and motor regions were engaged when she imagined that she was scratching her left cheek with her left plegic hand, whereas when she performed the same task with the SPL, additional left middle occipital areas were recruited. Moreover, comparison of responses induced by left cheek (subjectively feasible) versus right cheek scratching (reportedly unfeasible movement) with the SPL demonstrated significant activation in right somesthetic areas. INTERPRETATION: These findings demonstrate that intentional movements of a seen and felt SPL activate premotor and motor areas together with visual and sensory cortex, confirming its multimodal dimension and the reliability of the patient's verbal reports. This observation, interpreted for cortical deafferentation/disconnection caused by subcortical brain damage, constitutes a new but theoretically predictable entity among disorders of bodily awareness. PMID- 19557859 TI - Negative fluid-attenuated inversion recovery imaging identifies acute ischemic stroke at 3 hours or less. AB - OBJECTIVE: To evaluate the use of fluid-attenuated inversion recovery (FLAIR) imaging as surrogate marker of lesion age within the first 6 hours of ischemic stroke. METHODS: e analyzed FLAIR and diffusion-weighted imaging (DWI) sequences performed within 6 hours of symptom onset in 120 consecutive patients with ischemic stroke with known symptom onset. The visibility of acute ischemic lesions on FLAIR images was judged in two steps (on FLAIR alone and with knowledge of DWI) and compared with DWI. RESULTS: egative FLAIR in the case of positive DWI allocated ischemic lesions to a time window 3 hours or less with a high specificity (0.93) and a high positive predictive value (0.94), whereas sensitivity (0.48) and negative predictive value (0.43) were low. Lesion visibility on FLAIR images alone (35.6%) and with knowledge of DWI (62.5%) was lower than on DWI (97.1%). The sensitivity of FLAIR increased with increasing time from symptom onset from 27.0/50.0% 1 year) for the entire surgical population (n = 62). INTERPRETATION: MSI spike localization increases the chance that the seizure onset zone is sampled when patients undergo ICEEG for presurgical epilepsy evaluations. The clinical impact of this effect, improving diagnostic yield of ICEEG, should be considered in surgery candidates who do not have satisfactory indication of epilepsy localization from seizure semiology, electroencephalogram, and magnetic resonance imaging. PMID- 19557861 TI - A novel Nav1.7 mutation producing carbamazepine-responsive erythromelalgia. AB - OBJECTIVE: Human and animal studies have shown that Na(v)1.7 sodium channels, which are preferentially expressed within nociceptors and sympathetic neurons, play a major role in inflammatory and neuropathic pain. Inherited erythromelalgia (IEM) has been linked to gain-of-function mutations of Na(v)1.7. We now report a novel mutation (V400M) in a three-generation Canadian family in which pain is relieved by carbamazepine (CBZ). METHODS: We extracted genomic DNA from blood samples of eight members of the family, and the sequence of SCN9A coding exons was compared with the reference Na(v)1.7 complementary DNA. Wild-type Na(v)1.7 and V400M cell lines were then analyzed using whole-cell patch-clamp recording for changes in activation, deactivation, steady-state inactivation, and ramp currents. RESULTS: Whole-cell patch-clamp studies of V400M demonstrate changes in activation, deactivation, steady-state inactivation, and ramp currents that can produce dorsal root ganglia neuron hyperexcitability that underlies pain in these patients. We show that CBZ, at concentrations in the human therapeutic range, normalizes the voltage dependence of activation and inactivation of this inherited erythromelalgia mutation in Na(v)1.7 but does not affect these parameters in wild-type Na(v)1.7. INTERPRETATION: Our results demonstrate a normalizing effect of CBZ on mutant Na(v)1.7 channels in this kindred with CBZ responsive inherited erythromelalgia. The selective effect of CBZ on the mutant Na(v)1.7 channel appears to explain the ameliorative response to treatment in this kindred. Our results suggest that functional expression and pharmacological studies may provide mechanistic insights into hereditary painful disorders. PMID- 19557862 TI - Glycogen synthase kinase-3 beta and tau genes interact in Parkinson's and Alzheimer's diseases. PMID- 19557863 TI - Pigmentation as a clue to understanding Parkinson's disease and melanoma. PMID- 19557864 TI - Is reflex sympathetic dystrophy/complex regional pain syndrome type I a small fiber neuropathy? AB - Neurologist S. Weir Mitchell first described "causalgia" following wartime nerve injury, with its persistent distal limb burning pain, swelling, and abnormal skin color, temperature, and sweating. Similar post-traumatic symptoms were later identified in patients without overt nerve injuries after trauma. This was labeled reflex sympathetic dystrophy (RSD; now complex regional pain syndrome type I [CRPS-I]). The pathophysiology of symptoms is unknown and treatment options are limited. We propose that persistent RSD/CRPS-I is a post-traumatic neuralgia associated with distal degeneration of small-diameter peripheral axons. Small-fiber lesions are easily missed on examination and are undetected by standard electrophysiological testing. Most CRPS features-spreading pain and skin hypersensitivity, vasomotor instability, osteopenia, edema, and abnormal sweating are explicable by small-fiber dysfunction. Small fibers sense pain and temperature but also regulate tissue function through neuroeffector actions. Indeed, small-fiber-predominant polyneuropathies cause CRPS-like abnormalities, and pathological studies of nerves from chronic CRPS-I patients confirm small fiber-predominant pathology. Small distal nerve injuries in rodents reproduce many CRPS features, further supporting this hypothesis. CRPS symptoms likely reflect combined effects of axonal degeneration and plasticity, inappropriate firing and neurosecretion by residual axons, and denervation supersensitivity. The resulting tissue edema, hypoxia, and secondary central nervous system changes can exacerbate symptoms and perpetuate pathology. Restoring the interest of neurologists in RSD/CRPS should improve patient care and broaden our knowledge of small-fiber functions. PMID- 19557865 TI - White matter lesions volume and motor performances in the elderly. AB - OBJECTIVES: To investigate the cross-sectional and longitudinal associations between performance-based measures of motor function and volume of white matter lesions (WMLs), and to examine the influence of the localization of these lesions. METHODS: At baseline, motor performances (maximum walking speed, Tinetti gait and balance subscales) were assessed in 1,702 subjects aged 80 years or younger from the Dijon (France), France center of the Three-City study. Volumes of WMLs lesions (total, periventricular, deep) were measured using an automated method of tissue segmentation and quantification of lesion size. At 8-year follow up, walking speed was evaluated in 1,086 subjects. RESULTS: At baseline, mean and 95% confidence interval (CI) walking speed was lower in subjects with total volumes of WMLs >or=90th percentile (1.50 [1.45-1.55] m/s) than in subjects with lower volumes (1.56 [1.55-1.58] m/s; p = 0.004). Baseline total volumes of WMLs above the 90th percentile predicted walking speed decline during follow-up (odds ratio [95% CI] for having the greatest walking speed decline = 2.3 [1.3-4.1], p = 0.006). Moreover, high volumes of periventricular but not deep WMLs were associated with slower walking speed at baseline (p = 0.005) and over time (p = 0.001), and with lower Tinetti gait subscore (p = 0.02). INTERPRETATION: Our study shows a cross-sectional and longitudinal association between high total volumes of WMLs, in particular volumes above the 90th percentile, and impaired mobility. These associations were independent of several confounders, including cognition, and were mainly accounted for by volumes of periventricular WMLs. These findings support the hypothesis of a vascular contribution to motor decline in the elderly. PMID- 19557866 TI - Apolipoprotein E-dependent accumulation of Alzheimer disease-related lesions begins in middle age. AB - OBJECTIVE: To study the prevalence and age dependency of senile plaques (SP) and neurofibrillary tangles (NFT), the brain changes characteristic of Alzheimer disease (AD), and their association with apolipoprotein E (APOE) genotypes in a community-dwelling normal population. METHODS: This neuropathological study used both silver staining and A beta immunohistochemistry in brain tissue microarrays, including SP coverage and NFT counts from frontal cortex and hippocampus, and APOE genotyping, and was performed on a consecutive prospective series of 603 subjects (aged between 0 and 97 years) of an unselected population living outside of institutions. Cases were subjected to autopsy following sudden or unexpected out-of-hospital death, covering 22.1% of the mortality of Tampere, Finland and its surroundings. None died of AD, although 22 (3.7%) were demented and 10 (1.7%) had memory problems. RESULTS: Of the series, 30.8% had SP, and 42.1% had NFT; these occurred more commonly among females and showed a strong relationship with age. Both changes had already appeared at around 30 years of age, reaching an occurrence of almost 100% in the oldest. SP were more frequent in APOE epsilon 4 carriers compared with noncarriers in every age group except the oldest (>90 years). The difference was most evident during the ages 50 to 59 years, where 40.7% of epsilon 4-carriers had SP, compared with 8.2% in noncarriers (odds ratio, 8.39; 95% confidence interval, 2.55-27.62). The difference in NFT prevalence between APOE genotypes was not statistically significant in any age group. INTERPRETATION: The brain changes associated with AD may already begin developing early in middle age, especially among APOE epsilon 4 carriers. PMID- 19557867 TI - Fulminant JC virus encephalopathy with productive infection of cortical pyramidal neurons. AB - The polyomavirus JC (JCV) is the causative agent of progressive multifocal leukoencephalopathy and of JCV granule cell neuronopathy. We present a human immunodeficiency virus-negative patient who experienced development of multiple cortical lesions, aphasia, and progressive cognitive decline after chemotherapy for non-small-cell lung cancer. Brain biopsy and cerebrospinal fluid polymerase chain reaction demonstrated JCV, and she had a rapidly fatal outcome. Postmortem analysis showed diffuse cortical lesions and areas of necrosis at the gray-white junction. Immunostaining showed a productive JCV infection of cortical pyramidal neurons, confirmed by electron microscopy, with limited demyelination. This novel gray matter syndrome expands the scope of JCV clinical presentation and pathogenesis. PMID- 19557868 TI - Neuromuscular involvement in various types of Ehlers-Danlos syndrome. AB - OBJECTIVE: Ehlers-Danlos syndrome (EDS) is a clinically and genetically heterogeneous group of heritable connective tissue disorders characterized by joint hypermobility, skin hyperextensibility, and tissue fragility. Muscle involvement is plausible based on recently discovered interactions between muscle cells and extracellular matrix molecules; however, muscle symptoms are only sporadically reported. We designed a cross-sectional study to find out whether neuromuscular features are part of EDS. METHODS: Standardized questionnaires, physical examination, nerve conduction studies, electromyography, muscle ultrasound, and muscle biopsy were performed in 40 EDS patients with the vascular, classic, tenascin-X (TNX)-deficient type EDS, and hypermobility type of EDS caused by TNXB haploinsufficiency. RESULTS: Muscle weakness, myalgia, and easy fatigability were reported by the majority of patients. Mild-to-moderate muscle weakness (85%) and reduction of vibration sense (60%) were common. Nerve conduction studies demonstrated axonal polyneuropathy in five patients (13%). Needle electromyography myopathic features in nine patients (26%) and a mixed neurogenic-myopathic pattern in most (60%). Muscle ultrasound showed increased echo-intensity (48%) and atrophy (50%). Mild myopathic features were seen on muscle biopsy of five patients (28%). Overall, patients with the hypermobility type EDS caused by TNXB haploinsufficiency were least affected. INTERPRETATION: Mild-to-moderate neuromuscular involvement is common in various types of EDS, with a remarkable relation between residual TNX level and degree of neuromuscular involvement, compatible with a dose-effect relation. The findings of this study should increase awareness of neuromuscular symptoms in EDS patients and improve clinical care. They also point to a role of the extracellular matrix in muscle and peripheral nerve function. PMID- 19557869 TI - Antibodies produced by clonally expanded plasma cells in multiple sclerosis cerebrospinal fluid. AB - OBJECTIVE: Intrathecal IgG synthesis, persistence of bands of oligoclonal IgG, and memory B-cell clonal expansion are well-characterized features of the humoral response in multiple sclerosis (MS). Nevertheless, the target antigen of this response remains enigmatic. METHODS: We produced 53 different human IgG1 monoclonal recombinant antibodies (rAbs) by coexpressing paired heavy- and light chain variable region sequences of 51 plasma cell clones and 2 B-lymphocyte clones from MS cerebrospinal fluid in human tissue culture cells. Chimeric control rAbs were generated from anti-myelin hybridomas in which murine variable region sequences were fused to human constant region sequences. Purified rAbs were exhaustively assayed for reactivity against myelin basic protein, proteolipid protein, and myelin oligodendrocyte glycoprotein by immunostaining of transfected cells expressing individual myelin proteins, by protein immunoblotting, and by immunostaining of human brain tissue sections. RESULTS: Whereas humanized control rAbs derived from anti-myelin hybridomas and anti myelin monoclonal antibodies readily detected myelin antigens in multiple immunoassays, none of the rAbs derived from MS cerebrospinal fluid displayed immunoreactivity to the three myelin antigens tested. Immunocytochemical analysis of tissue sections from MS and control brain demonstrated only weak staining with a few rAbs against nuclei or cytoplasmic granules in neurons, glia, and inflammatory cells. INTERPRETATION: The oligoclonal B-cell response in MS cerebrospinal fluid is not targeted to the well-characterized myelin antigens myelin basic protein, proteolipid protein, or myelin oligodendrocyte glycoprotein. PMID- 19557870 TI - Oxidative stress in SEPN1-related myopathy: from pathophysiology to treatment. AB - OBJECTIVE: Mutations of the selenoprotein N gene (SEPN1) cause SEPN1-related myopathy (SEPN1-RM), a novel early-onset muscle disorder formerly divided into four different nosological categories. Selenoprotein N (SelN) is the only selenoprotein involved in a genetic disease; its function being unknown, no treatment is available for this potentially lethal disorder. Our objective was to clarify the role of SelN and the pathophysiology of SEPN1-RM to identify therapeutic targets. METHODS: We established and analyzed an ex vivo model of SelN deficiency using fibroblast and myoblast primary cultures from patients with null SEPN1 mutations. DCFH assay, OxyBlot, Western blot, Fura-2, and cell survival studies were performed to measure intracellular oxidant activity, oxidative stress markers, calcium handling, and response to exogenous treatments. RESULTS: SelN-depleted cells showed oxidative/nitrosative stress manifested by increased intracellular oxidant activity (reactive oxygen species and nitric oxide) and/or excessive oxidation of proteins, including the contractile proteins actin and myosin heavy chain II in myotubes. SelN-devoid myotubes showed also Ca(2+) homeostasis abnormalities suggesting dysfunction of the redox-sensor Ca(2+) channel ryanodine receptor type 1. Furthermore, absence of SelN was associated with abnormal susceptibility to H(2)O(2)-induced oxidative stress, demonstrated by increased cell death. This cell phenotype was restored by pretreatment with the antioxidant N-acetylcysteine. INTERPRETATION: SelN plays a key role in redox homeostasis and human cell protection against oxidative stress. Oxidative/nitrosative stress is a primary pathogenic mechanism in SEPN1-RM, which can be effectively targeted ex vivo by antioxidants. These findings pave the way to SEPN1-RM treatment, which would represent a first specific pharmacological treatment for a congenital myopathy. PMID- 19557872 TI - The specificity of oligoclonal bands in multiple sclerosis. PMID- 19557874 TI - Alzheimer disease: multiple causes, multiple effects of apolipoprotein E4, and multiple therapeutic approaches. PMID- 19557875 TI - Understanding the role of natural killer cell receptors and their human leukocyte antigen ligands in multiple sclerosis. PMID- 19557876 TI - B vitamins may protect against macular degeneration. PMID- 19557877 TI - Cytoprotection against hydrogen peroxide-induced cell death in cultured mouse mesangial cells by erigeroflavanone, a novel compound from the flowers of Erigeron annuus. AB - Hyperglycemia-induced oxidative stress has been suggested as a mechanism underlying diabetic complications. Oxidative stress triggers cell death in various cell types, including glomerular mesangial cells which play important roles in diabetic nephropathy. In the present study, we investigated the potential cytoprotective effect of erigeroflavanone, a novel flavanone derivative from the flowers of Erigeron annuus, in cultured mouse mesangial cells using hydrogen peroxide (H2O2) as an oxidative stress inducer. Our data show that hydrogen peroxide induced a decrease in cell viability that was attenuated by erigeroflavanone. Hydrogen peroxide treatment increased formation of dichlorofluorescein (DCF)-sensitive intracellular reactive oxygen species (ROS). This enhanced ROS formation was significantly reduced by pretreatment with erigeroflavanone in a dose-dependent manner. Hydrogen peroxide treatment also induced phosphorylation of the mitogen-activated protein kinases (MAPKs), c-Jun terminal kinase (JNK), extracellular-regulated kinase (ERK) and p38, and activated caspase-3. Pretreatment with erigeroflavanone inhibited hydrogen peroxide-induced activation of MAPKs and caspase-3. From these data we conclude that erigeroflavanone provides a protective effect against oxidative stress induced cell death in mesangial cells that is associated with its antioxidant action and inhibition of MAPKs and caspase-3. These results suggest that erigeroflavanone has potential as a therapeutic agent in the treatment of renal diabetic complications. PMID- 19557878 TI - Low-dose of tacrolimus favors the induction of functional CD4(+)CD25(+)FoxP3(+) regulatory T cells in solid-organ transplantation. AB - It has been reported that low-dose of tacrolimus (Tac) is advantageous for the long-term allograft function and survival when compared with standard-dose of tacrolimus. However, the underlying mechanism is not known and remains to be elucidated. CD4(+)CD25(+)FoxP3(+) regulatory T cells (Tregs) play a key role in the induction and maintenance of transplantation tolerance. We studied whether low-dose of Tac would favor the induction of donor-specific Tregs. We found that in all transplant recipients treated with low-dose of Tac (target trough level of 3 to 7 ng/ml), CD4(+)CD25(+)FoxP3(+) Tregs were induced and expanded in the periphery and accumulated in the allograft after transplantation, and they retained their suppressive efficacy in vitro. When studied in vitro, we found that high concentration of Tac significantly decreased the induction of FoxP3 expression in mixed lymphocyte reactions (MLR) when compared with low concentration of Tac. Taken together, these data imply that in solid-organ transplantation the minimization of Tac might be beneficial and favors the induction of donor-specific Tregs maintaining transplantation tolerance to alloantigen. PMID- 19557879 TI - Immunoregulatory Cordyceps sinensis increases regulatory T cells to Th17 cell ratio and delays diabetes in NOD mice. AB - Cordyceps sinensis (CS) is a parasitic fungus, and it has been used widely in traditional Chinese medicines (TCM) for centuries. Many studies have shown that CS has immunoregulatory activity in many disease models, but the underlying mechanism remains elusive. We studied whether CS could suppress the onset of diabetes by altering T lymphocyte subsets in non-obese diabetic (NOD) mice. We found that the onset of type 1 diabetes in NOD mice was associated with an imbalance of CD4(+)CD25(+)FoxP3(+) regulatory T (Treg) cells and IL-17 producing Th17 cells. Oral administration of CS resulted in reduction in the overall incidence of diabetes, and this was due to an increase in the ratio of Treg cells to Th17 in the spleen and pancreatic lymph nodes (PLNs). Taken together, these data imply that CS is able to modulate Treg to Th17 cell ratio in vivo, thus contributing to the inhibition of diabetes. PMID- 19557880 TI - Commentaries on viewpoint: perception of effort during exercise is independent of afferent feedback from skeletal muscles, heart and lungs. PMID- 19557881 TI - Commentaries on viewpoint: perception of effort during exercise is independent of afferent feedback from skeletal muscles, heart,and lungs. PMID- 19557882 TI - Commentaries on viewpoint: perception of effort during exercise is independent of afferent feedback from skeletal muscles, heart,and lungs. PMID- 19557883 TI - Commentaries on viewpoint: perception of effort during exercise is independent of afferent feedback from skeletal muscles, heart,and lungs. PMID- 19557884 TI - Deep sequencing maps the maize epigenomic landscape. PMID- 19557885 TI - Clinical study or research activity? PMID- 19557886 TI - Arrhythmogenic right ventricular cardiomyopathy. PMID- 19557887 TI - Ozone exposure and mortality. PMID- 19557888 TI - Growth in care provided by hospitalists. PMID- 19557889 TI - Medical evaluation before electroconvulsive therapy. PMID- 19557890 TI - Ethical and scientific implications of the globalization of clinical research. PMID- 19557891 TI - Carcinoma en cuirasse of the breast. PMID- 19557892 TI - Osteoclasts and arthritis. PMID- 19557893 TI - Special issue dedicated to Dr. Jurg Graf. Proceedings of a symposium on cell biology of the hepatobiliary system, October 2007, Vienna, Austria. PMID- 19557894 TI - Pharmacoeconomic evaluations of antifungal therapies. AB - Invasive fungal infections are a growing source of morbidity and mortality in the United States and are associated with excess hospital stay and costs. Although clinicians now have more effective and safe treatment options for invasive candidiasis, evidence confirming the pharmacoeconomic benefits of newer treatment options such as the echinocandins is scarce as few rigorous pharmacoeconomic studies have been completed in the arena of invasive fungal infections. Beyond the cost of drug acquisition, costs associated with ineffective treatment and significant drug-related side-effects must be factored into pharmacoeconomic analysis. Additional administrative costs may need to be considered when a formulary medication switch is undertaken, including those involved with re education of clinicians unfamiliar with the newer product. PMID- 19557895 TI - The management of thymoma: a systematic review and practice guideline. AB - INTRODUCTION: Thymoma is a rare tumor for which there is little randomized evidence to guide treatment. Because of the lack of high-quality evidence, a formal consensus-based approach was used to develop recommendations on treatment. METHODS: A systematic refview of the literature was performed. Recommendations were formed from available evidence and developed through a two-round modified Delphi consensus approach. RESULTS: The treatment recommendations are summarized as follows: Stage I--complete resection of the entire thymus without neoadjuvant or adjuvant therapy. Stage II--complete resection of the entire thymus with consideration of adjuvant radiation for high-risk tumors. Stage IIIA--surgery either initially or after neoadjuvant therapy, or surgery followed by adjuvant therapy. Stage IIIB--treatment may include a combination of chemotherapy, radiation, and/or surgery, or if technically possible, surgery in combination with chemoradiotherapy (concurrent cisplatin based). For bulky tumors, consideration should be given to sequential chemotherapy followed by radiation. Stage IVA--as per stage III, with surgery only if metastases can be resected. Stage IVB--treatment on an individual case basis (no generic recommendations). Recurrent disease--consider surgery, radiation, and/or chemoradiation. Chemoradiation should be considered in all medically inoperable and technically inoperable patients. CONCLUSION: Consensus was achieved on these recommendations, which serve to provide practical guidance to the physician treating this rare disease. PMID- 19557896 TI - Beefed up. PMID- 19557897 TI - Fence protection progress. PMID- 19557898 TI - Kept out. PMID- 19557899 TI - Cheek by jowl. PMID- 19557900 TI - Island refuge. PMID- 19557901 TI - Jon Driver. PMID- 19557902 TI - Pancreatic regenerating gene I and acinar cell differentiation: influence on cellular lineage. AB - OBJECTIVES: Pancreatic regenerating gene I (reg I) has been implicated in cellular differentiation. Acinar cells can transdifferentiate into other pancreatic-derived cells, and we postulated that changes in intracellular levels of reg I would affect the state of differentiation. METHODS: We transfected AR42J cells with a plasmid containing the entire coding sequence of reg I and isolated clones with complementary DNA in sense (SS) or antisense (AS) orientation. Levels of messenger RNA (mRNA) and protein expression were examined by Western blotting and real-time polymerase chain reaction. RESULTS: Expression of reg I was confirmed in SS or AS clones. AR42J transfected with SS demonstrated more acinarlike phenotype, whereas those transfected with AS showed a less differentiated state. Specifically, amylase mRNA and protein levels increased in SS cells, whereas AS cells showed increased pancreatic and duodenal homeobox 1 (Pdx1) and insulin mRNAs and cytokeratin protein. Conversely, cytokeratin and Pdx1 were depressed in SS cells. CONCLUSIONS: These data demonstrate that in acinar cells, reg I overexpression is linked to acinar cell differentiation, whereas inhibition of reg I leads to beta cell and possibly ductal phenotype. Reg I expression in acinar cells is important in maintaining pancreatic cell lineage, and when decreased, cells can dedifferentiate and move toward becoming other pancreatic cells. PMID- 19557903 TI - The moral is the story. AB - When religious systems take on secular partners, they can find themselves in difficult situations regarding their ethical obligations. In Denver, a Roman Catholic system is co-sponsor of hospitals that perform abortions and other services. But the battle to unwind the business relationship has been knocked off track by resistance from the hospital management company. Exempla considers its "dual heritage" a strength, says CEO Jeff Selberg, left. PMID- 19557904 TI - Dem leaders hash out reform ... as GOP lawmakers say they're being left out. PMID- 19557906 TI - Charitable definitions. HFMA takes strict stance on charity care. PMID- 19557905 TI - Some wary of RAC's cousin. Exec: MIC makes RAC seem like child's play. PMID- 19557907 TI - Borrowing bounces back. Bond issues on pace to topple '08 figures. PMID- 19557908 TI - Reform imperatives. Three strategies that anticipate Washington's work. PMID- 19557909 TI - Making a comeback. While interest rates are still more costly and financings more complex, credit continues to loosen. PMID- 19557910 TI - Does science education need the history of science? PMID- 19557911 TI - Care Quality Commission. Watchdog chief sets out her vision. PMID- 19557912 TI - Quality of care. Who let standards fall so low at Mid Staffordshire? PMID- 19557913 TI - Simon Stevens on how to help doctors think. PMID- 19557914 TI - Peak planning is cold comfort. PMID- 19557915 TI - Data briefing. The north-south alcohol divide. PMID- 19557916 TI - Obesity management. The fat of the land. AB - Sixty per cent of men and 50 per cent of women in the UK could be obese by 2050. The government wants to reduce the number of obese and overweight children by 2020. Obesity management is far less embedded in general healthcare than smoking cessation. PMID- 19557917 TI - Acute care. Big break for the clinical coders. PMID- 19557918 TI - Joint coaching. Clash of the titan personalities. PMID- 19557919 TI - Patient safety. Penalties for infections may put trust in the red. PMID- 19557920 TI - Foundation trusts. Corporate flair to the fore in brave new world. PMID- 19557921 TI - Noel Plumridge on foundation trusts. PMID- 19557922 TI - Forget fairness, this is the future. PMID- 19557923 TI - Dementia. Improve care through a pathway to early diagnosis. AB - The number of people with dementia is set to rise to 1.4 million over the next 30 years. The National Dementia Strategy's objectives are to improve awareness, diagnosis, intervention and care. The strategy's objectives outline the need for a pathway of care that delivers specialist assessment, treatment and support with capacity to see all new cases in an area. The initial five year strategy implementation was backed by pound 150m over the first two years. Cost effectiveness analysis indicates that enhanced memory services can improve care quality and save hundreds of millions of pounds over 10 years. PMID- 19557924 TI - Mixed accommodation. Deliver dignity or pay for mixed sex wards. PMID- 19557925 TI - Foundation trusts. Breaking free. AB - The foundation trust model was strongly debated when it was first introduced five years ago. Those involved in creating them expected the model to give hospitals more freedom to innovate. PMID- 19557931 TI - Differential effect of Shenmai injection, a herbal preparation, on the cytochrome P450 3A-mediated 1'-hydroxylation and 4-hydroxylation of midazolam. AB - Shenmai injection (SMI), one of the most popular herbal preparations, is widely used for the treatment of coronary atherosclerotic cardiopathy and viral myocarditis. The purpose of this study was to investigate the effect of Shenmai injection (SMI) on the CYP3A-mediated metabolism of midazolam (MDZ). The present study demonstrated that SMI could significantly inhibit MDZ 4-hydroxylation but activate its 1'-hydroxylation in human liver microsomes (HLMs), rat liver microsomes (RLM) and recombinant human CYP3A4 and CYP3A5. The opposing effect of SMI was characterized by the kinetic change of increasing Vmax/Km for MDZ 1' hydroxylation and decreasing Vmax/Km for MDZ 4-hydroxylation in HLM and RLM. The presence of SMI enhanced the inhibition of ketoconazole on MDZ 4-hydroxylation but weakened or reversed its inhibition on MDZ 1'-hydroxylation in HLM. After single or multiple pretreatment with SMI, the ratios of AUC(4-OH MDZ)/AUC(MDZ) in rats were significantly decreased, while the ratios of AUC(1'-OH MDZ)/AUC(MDZ) were increased. Among the major components in SMI, total ginsenoside (TG), ophiopogon total saponins (OTS), ophiopogon total flavone (OTF), ginsenoside Rd, ophiopogonin D and ophiopogonone A exhibited significant inhibition on both 4 hydroxylation and 1'-hydroxylation of MDZ in HLM and RLM, while no activation on MDZ metabolism was observed in the presence of these major constituents alone or together. To further explore the responsible components, 3 mL of SMI was loaded on a solid phase extraction (SPE) C18 cartridge and then separated by different concentrations of methanol. The fractions eluted with 60% and 90% methanol both showed significant activation on MDZ 1'-hydroxylation in HLM, but the fraction eluted with 30% methanol had no such effect. The results indicated that the activation of SMI on MDZ 1'-hydroxylation might be mainly resulted from the lipid soluble components in SMI. PMID- 19557932 TI - Corollary discharges and perception of effort are dissociated during repeated sprints. PMID- 19557933 TI - Commentaries on viewpoint: perception of effort during exercise is independent of afferent feedback from skeletal muscles, heart,and lungs. PMID- 19557934 TI - Commentaries on viewpoint: perception of effort during exercise is independent of afferent feedback from skeletal muscles, heart,and lungs. PMID- 19557935 TI - The integration of afferent feedback on the perception of effort cannot be dismissed. PMID- 19557936 TI - The perception of effort: mind over muscle? PMID- 19557937 TI - Echinocandins: pharmacokinetic and therapeutic issues. AB - Invasive infections caused by Candida species are a major concern today due to increasing numbers of at-risk patients and high rates of mortality, as well as increased length of hospital stays and healthcare costs associated with these infections. An additional concern is the current trend towards greater numbers of infections caused by species or strains that are resistant to traditional antifungal agents, such as fluconazole or other triazoles. Echinocandins are the newest class of antifungals. They have a unique mechanism of action, and exhibit activity against a broad range of Candida species and strains, including those resistant to azoles or polyenes. Few studies have directly compared the three approved echinocandins (caspofungin, micafungin, and anidulafungin) for efficacy, but the existing data have not suggested major differences to date. All the echinocandins possess excellent tolerability and safety, and although there are some pharmacokinetic differences, they are relatively small and generally do not influence drug selection. Consequently, echinocandins are now considered by most experts to be the first-line treatment of invasive candidiasis in critically ill patients. Caspofungin was the first approved member of the class; it has the most available data and the most indications of the echinocandins. PMID- 19557938 TI - Treating HIV infection immediately might stall immune decay. PMID- 19557939 TI - Merck aims for male Gardasil. PMID- 19557940 TI - International disease surveillance. International disease monitoring, January to March 2008. PMID- 19557941 TI - [Osteoarthritis epidemics in Denmark?]. PMID- 19557942 TI - New influenza A (H1N1) virus: WHO guidance on public health measures, 11 June 2009. PMID- 19557943 TI - Drug resistance in leprosy: reports from selected endemic countries. PMID- 19557944 TI - [How many (senior)physicians to write a case report?]. PMID- 19557945 TI - Health politics, policy and law. Editor's note. PMID- 19557947 TI - Self care abilities of patients in a referral hospital of North India. AB - A descriptive study was undertaken to assess the self-care abilities among the admitted patients at Nehru Hospital, PGIMER Chandigarh during the year 2004. Various self-care abilities studied were regarding feeding, bathing, dressing and toileting. Out of total 650 patients included in the study, 394 (45.2%) had feeding deficit, 291 (44.84%) had bathing deficit, 238 (36.73%) had dressing deficit and 197 (31.88%) had the deficit of toileting. As per the severity of deficit, mild deficit was present in 121 (19.6%), moderate in 91 (14.70%) and severe deficit was present in 59 (9.6%) of the patients. PMID- 19557948 TI - Future implications for nursing practice. PMID- 19557949 TI - AIDS -- related stigma. PMID- 19557950 TI - Competency and competence. PMID- 19557951 TI - Potential roles of military-specific response to natural disasters -- analysis of the rapid deployment of a mobile surgical team to the 2007 Peruvian earthquake. AB - The August 2007 earthquake in Peru resulted in the loss of critical health infrastructure and resource capacity. A regionally located United States Military Mobile Surgical Team was deployed and operational within 48 hours. However, a post-mission analysis confirmed a low yield from the military surgical resource. The experience of the team suggests that non-surgical medical, transportation, and logistical resources filled essential gaps in health assessment, evacuation, and essential primary care in an otherwise resource-poor surge response capability. Due to an absence of outcomes data, the true effect of the mission on population health remains unknown. Militaries should focus their disaster response efforts on employment of logistics, primary medical care, and transportation/evacuation. Future response strategies should be evidence-based and incorporate a means of quantifying outcomes. PMID- 19557952 TI - Military medical assistance following natural disasters: refining the rapid response. PMID- 19557953 TI - Mississippi's infectious disease hotline: a surveillance and education model for future disasters. AB - INTRODUCTION: The potential for outbreaks of epidemic disease among displaced residents was a significant public health concern in the aftermath of Hurricane Katrina. In response, the Mississippi Department of Health (MDH) and the American Red Cross (ARC) implemented a novel infectious disease surveillance system, in the form of a telephone "hotline", to detect and rapidly respond to health threats in shelters. METHODS: All ARC-managed shelters in Mississippi were included in the surveillance system. A symptom-based, case reporting method was developed and distributed to shelter staff, who were linked with MDH and ARC professionals by a toll-free telephone service. Hotline staff investigated potential infectious disease outbreaks, provided assistance to shelter staff regarding optimal patient care, and helped facilitate the evaluation of ill evacuees by local medical personnel. RESULTS: Forty-three shelters sheltering 3,520 evacuees participated in the program. Seventeen shelters made 29 calls notifying the hotline of the following cases: (1) fever (6 cases); (2) respiratory infections (37 cases); (3) bloody diarrhea (2 cases); (4) watery diarrhea (15 cases); and (5) other, including rashes (33 cases). Thirty-four of these patients were referred to a local physician or hospital for further diagnosis and disease management. Three cases of chickenpox were identified. No significant infectious disease outbreaks occurred and no deaths were reported. CONCLUSIONS: The surveillance system used direct verbal communication between shelter staff and hotline managers to enable more rapid reporting, mapping, investigation, and intervention, far beyond the capabilities of a more passive or paper-based system. It also allowed for immediate feedback and education for staff unfamiliar with the diseases and reporting process. Replication of this program should be considered during future disasters when health surveillance of a large, disseminated shelter population is necessary. PMID- 19557954 TI - Mustard gas or sulfur mustard: an old chemical agent as a new terrorist threat. AB - Sulfur mustard is a member of the vesicant class of chemical warfare agents that causes blistering to the skin and mucous membranes. There is no specific antidote, and treatment consists of systematically alleviating symptoms. Historically, sulfur mustard was used extensively in inter-governmental conflicts within the trenches of Belgium and France during World War I and during the Iran Iraq conflict. Longitudinal studies of exposed victims show that sulfur mustard causes long-term effects leading to high morbidity. Given that only a small amount of sulfur mustard is necessary to potentially cause an enormous number of casualties, disaster-planning protocol necessitates the education and training of first-line healthcare responders in the recognition, decontamination, triage, and treatment of sulfur mustard-exposed victims in a large-scale scenario. PMID- 19557955 TI - Crowd behavior at mass gatherings: a literature review. AB - Gaining an understanding of crowd behavior is important in supporting timely and appropriate crowd management principles in the planning and provision of emergency services at mass gatherings. This paper provides a review of the current understanding of the psychological factors of a crowd within the psychosocial domain as they apply to mass-gathering settings. It can be concluded from this review that there is a large theory-practice gap in relation to crowd psychology and the mass-gathering setting. The literature has highlighted two important elements of crowd behavior-there must be a "seed" and people must engage. Understanding these behaviors may provide opportunities to change crowd behavior outcomes. PMID- 19557956 TI - Application of outcome measures in international humanitarian aid: comparing indices through retrospective analysis of corrective surgical care cases. AB - It is common for international organizations to provide surgical corrective care to vulnerable populations in developing countries. However, a current worsening of the overall surgical burden of disease in developing countries reflects an increasing lack of sufficient numbers of trained healthcare personnel, and renders outside volunteer assistance more desirable and crucial than ever. Unfortunately, program evaluation and monitoring, including outcome indices and measures of effectiveness, is not measured commonly. In 2005, Operation Smile International implemented an electronic medical record system that helps monitor a number of critical indices during surgical missions that are essential for quality assurance reviews. This record system also provided an opportunity to retrospectively evaluate cases from previous missions. Review of data sets from >8,000 cases in 2005 and 2006 has provided crucial information regarding the priority of surgery, perioperative and operative complications, and surgical program development. The most common procedure provided was unilateral cleft lip repair, followed closely by cleft palate. A majority of these interventions occurred for patients who were older than routinely provided for in the western world. The average child treated had an age:weight ratio at or below the [US] Centers for Disease Control and Prevention (CDC) 50th percentile, with a small percentage falling below the CDC 20th percentile. A majority of children had acceptable levels of hemoglobin, but the relative decreased age:weight ratio nonetheless can reflect mild malnutrition. Complications requiring medical intervention were seen in 1.2% of cases in 2005 and 1.0% in 2006. Thirty percent were reported as anesthesia complications, and 61% reported as surgical complications. One death was reported, but occurred after discharge outside the perioperative period. Complication rates are similar to rates reported in the US and UK and emphasizes the importance of standardization with uniform indices to compare quality performance and equity of care. This study offers an important example of the importance of collecting, analyzing, and reporting measures of effectiveness in all surgical settings. PMID- 19557957 TI - Critical challenges ahead in bioterrorism preparedness training for clinicians. AB - PURPOSE: A survey was distributed to determine physicians' confidence levels in recognizing potential Category-A bioterrorism disease threats (e.g., smallpox, anthrax), preferred means of obtaining continuing medical education (CME) credits, and their knowledge of the Connecticut Department of Public Health's (DPH) disease reporting requirements. METHODS: Surveys were mailed to all physicians in the three-hospital Yale New Haven Health (YNHH) System (2,174) from January to March 2004; there were 820 respondents for a 37.7% response rate. RESULTS: A total of 71% of physicians indicated that they were "not confident" that they could recognize five of the infectious agents named; they had higher confidence rates for smallpox (48.8%). Infectious diseases and emergency medicine physicians had the highest rates of confidence. Seventy-eight percent of physicians indicated conferences and lectures as their preferred CME learning modality. Nearly 72% of physicians reported a low familiarity with the DPH reporting requirements. DISCUSSION: The results highlighted the breadth of perceived weaknesses among clinicians from disease recognition to reporting incidents, which signifies the need for greater training in these areas. As clinicians themselves emphasized their lack of skills and knowledge in this area, there should be a rapid development and dissemination of problem-based learning CME courses in bioterrorism preparedness. PMID- 19557958 TI - Resilience and challenges among staff of gulf coast nursing homes sheltering frail evacuees following Hurricane Katrina, 2005: implications for planning and training. AB - PURPOSE: The purpose of this study was to: (1) explore experiences and responses of staff in caring for sheltered, frail, Hurricane Katrina evacuees; and (2) identify how planning and training can be enhanced for staff who may care for frail older populations during and after disasters. METHODS: Individual, in person, semi-structured interviews were conducted with 38 staff members in four nursing homes in Mississippi, sheltering 109 evacuees in November 2005, nine weeks after Hurricane Katrina. Twenty-four were direct care staff, including certified nursing assistants, licensed nurses, dietary aides, and social workers; 14 were support staff, including maintenance and business managers. The number interviewed in each nursing home averaged 9.5 (range 6-15). Using a discussion guide and focusing on their experiences caring for nursing home evacuees, staff were asked to describe: (1) experiences; (2) problems; (3) what helped; and (4) what was learned. Data were processed using grounded theory and thematic analysis. Responses of direct care staff differed in emphasis from those of support staff in several areas; responses from these groups were analyzed separately and together. Three of the researchers identified recurring themes; two organized themes conceptually. RESULTS: Staff emphasized providing emotional reassurance to evacuees as well as physical care. Many described caring for evacuees as "a blessing," saying the experience helped them bond with residents, evacuees, and other staff. However, caring for evacuees was difficult because staff members were extremely anxious and in poor physical condition after an arduous evacuation. Challenges included communicating with evacuees' families, preventing dehydration, lack of personal hygiene supplies, staff exhaustion, and emotional needs of residents, evacuees, and staff. Teamwork, community help, and having a well-organized disaster plan, extra supplies, and dependable staff helped personnel cope with the situation. CONCLUSIONS: Staff of nursing homes that sheltered Katrina evacuees demonstrated resilience in the disaster's aftermath. Many placed the well-being of residents as their first priority. Results underscore the importance of planning, teamwork, and adequate supplies and staffing. Training for long-term care staff should emphasize providing emotional support as well as physical care for residents and evacuees during and following disasters. Nurses, social workers, and other staff members responsible for promoting emotional well-being for nursing home residents should be prepared to respond to disasters. PMID- 19557959 TI - Laryngeal tube and intubating laryngeal mask insertion in a manikin by first responder trainees after a short video-clip demonstration. AB - INTRODUCTION: This study was performed in the Finnish Defense Forces to assess the potential applicability and value of short video clips as educational material to teach advanced airway management and as the first means of introducing the use of a laryngeal tube (LT) or an intubating laryngeal mask (ILMA) to inexperienced, military, first-responder trainees with no prior hands on experience. METHODS: The 60 non-commissioned medical officers participating in this study were randomly assigned into one of two groups: the LT- and the ILMA group. After viewing the video clips, the trainees were required to perform 10 consecutive, successful insertions of the given instrument into a manikin. The number and duration of the attempts required prior to the 10 consecutive successful insertions were measured. RESULTS: The goal of 10 consecutive successful insertions was attained by all 30 subjects in the LT-group, and by 27 of 29 subjects in the ILMA-group with a maximum of 30 attempts. Improvement in the ease and speed of insertion was evident between the first and last consecutive insertions in both groups. CONCLUSIONS: "Satisfactory" to "good" skill levels are achieved with the applied video-clip demonstration method, even in inexperienced first-responder trainees lacking previous hands on experience. PMID- 19557960 TI - Poison control centers' role in glow product-related outbreak detection: implications for comprehensive surveillance system. AB - INTRODUCTION: The development of syndromic surveillance systems to detect bioterrorist attacks and emerging infectious diseases has become an important and challenging goal to many governmental agencies and healthcare authorities. This study utilized the sharp increase of glow product-related calls to demonstrate the utility of poison control data for early detection of potential outbreaks during the week of Halloween in 2007. METHODS: A review was conducted of the electronic records of exposures reported to the New Jersey Poison Information and Education System (NJPIES) Poison Control Hotline from 2002 through 2007 with generic code number 0201027 (glow products) set by the American Association of Poison Control Centers (AAPCC). Key information such as age, gender, time of the call, exposure reason, clinical effects, and medical outcomes along with telephone number, zip code, and county location were used in the analyses to determine the extent of the outbreak. RESULTS: Analyses included a total of 139 glow product-related calls during the week of Halloween in 2007 with a single-day high of 59 calls on Halloween Day. More than 90% of the glow product exposures were in children 1-10 years of age. The glow product-related calls on Halloween Day increased from 14 calls in 2002 to 59 calls in 2007, a 321% increase during a six-year period. CONCLUSIONS: Poison control centers in the United States are equipped with a unique and uniform input data collection system -- the National Poison Data System -- that provides an important data source in the development of a comprehensive surveillance system for early outbreak detection. PMID- 19557961 TI - Medical society's blueprint for a successful community response to emergency preparedness. AB - It is clear from disaster evaluations that communities must be prepared to act independently before government agencies can cope with the early ramifications of disasters. In response to devastation to the borough of Staten Island, New York in the wake of 11 September 2001, the Richmond County Medical Society established a structure to incorporate community needs and institutions to work together for the common good. A program that brings together two hospital systems, nursing homes, emergency medical services, and the Office of Emergency Management physician leadership in a meaningful way now is in place. This approach has improved the disaster preparedness of Staten Island and demonstrated how the Medical Society can provide leadership in disaster preparedness and serve as a conduit for communication amongst entities that normally do not communicate. PMID- 19557962 TI - Non-doctors as trauma surgeons? A controlled study of trauma training for non graduate surgeons in rural Cambodia. AB - INTRODUCTION: Due to the accelerating global epidemic of trauma, efficient and sustainable models of trauma care that fit low-resource settings must be developed. In most low-income countries, the burden of surgical trauma is managed by non-doctors at local district hospitals. OBJECTIVE: This study examined whether it is possible to establish primary trauma surgical services of acceptable quality at rural district hospitals by systematically training local, non-graduate, care providers. METHODS: Seven district hospitals in the most landmine-infested provinces of Northwestern Cambodia were selected for the study. The hospitals were referral points in an established prehospital trauma system. During a four-year training period, 21 surgical care providers underwent five courses (150 minutes total) focusing on surgical skills training. In-hospital trauma deaths and post-operative infections were used as quality-of care indicators. Outcome indicators during the training period were compared against pre-intervention data. RESULTS: Both the control and treatment populations had long prehospital transport times (three hours) and were severely injured (median Injury Severity Scale Score = 9). The in-hospital trauma fatality rate was low in both populations and not significantly affected by the intervention. The level of post-operative infections was reduced from 22.0% to 10.3% during the intervention (95% confidence interval for difference 2.8-20.2%). The trainees' self-rating of skills (Visual Analogue Scale) before and after the training indicated a significantly better coping capacity. CONCLUSIONS: Where the rural hospital is an integral part of a prehospital trauma system, systematic training of non-doctors improves the quality of trauma surgery. Initial efforts to improve trauma management in low-income countries should focus on the district hospital. PMID- 19557963 TI - The lasting legacy of war: epidemiology of injuries from landmines and unexploded ordnance in Afghanistan, 2002-2006. AB - INTRODUCTION: Due to several decades of armed conflict and civil unrest, Afghanistan is one of the countries most affected by landmines and unexploded ordnance worldwide. OBJECTIVE: The study was performed to assess the magnitude of injuries due to landmines and unexploded ordnance in Afghanistan during 2002-2006 and to describe epidemiological patterns and potential risk factors for these events. METHODS: Surveillance data including 5,471 injuries caused by landmines and unexploded ordnance in Afghanistan during 2002-2006 were analyzed. The International Committee of the Red Cross collects data on such injuries from 490 reporting health facilities and volunteers throughout the country. These surveillance data were used to describe injury trends, victim demographics, injury types, risk behaviors, and explosive types related to landmine and unexploded ordnance accidents. RESULTS: The largest number of injuries (1,706) occurred in 2002. The number declined sharply to 1,049 injuries in 2003, and remained relatively stable with slight decline thereafter. Overall, 92% of victims were civilians, 91% were males, and 47% were children <18 years of age. The case-fatality ratio was 17%. Approximately 50% of all injuries were caused by unexploded ordnance and 42% by landmines. Among children, 65% of injuries were caused by unexploded ordnance and only 27% by landmines, whereas in adults, most injuries (56%) were caused by landmines. The most common risk behaviors among children were tending animals, playing, and tampering with explosive devices. In adults, most common risk behaviors were traveling, performing activities of economic necessity, and tampering with explosives. Twenty-eight percent of the surviving victims who received mine awareness training and 2% of those who did not receive such training reported that the area where event occurred was marked. CONCLUSIONS: The large number of injuries and high proportion of child victims suggest that clearance and risk education activities fall short of achieving their goals, and must be substantially improved or expanded. Especially concerning is the high proportion of injuries caused by unexploded ordnance, and the high number of injuries sustained while tampering with explosive devices. Because unexploded ordnance is more visible than are landmines, and ordnance contaminated areas are cheaper to clear than are minefields, these injuries are highly preventable and should be a priority for clearance and risk education efforts. PMID- 19557964 TI - Injury perceptions of bombing survivors--interviews from the Oklahoma City bombing. AB - INTRODUCTION: Bombings, including the 1995 Oklahoma City bombing, remain an important public health threat. However, there has been little investigation into the impressions of injury risk or protective factors of bombing survivors. OBJECTIVE: This study analyzes Oklahoma City bombing survivors' impressions of factors that influenced their risk of injury, and validates a hazard timeline outlining phases of injury risk in a building bombing. METHODS: In-depth, semi structured interviews were conducted within a sample of Oklahoma City bombing survivors. Participants included 15 injured and uninjured survivors, who were located in three buildings surrounding the detonation site during the attack. RESULTS: Risk factor themes included environmental glass, debris, and entrapment. Protective factors included knowledge of egress routes, shielding behaviors to deflect debris, and survival training. Building design and health status were reported as risk and protective factors. The hazard timeline was a useful tool, but should be modified to include a lay rescue phase. The combination of a narrative approach and direct questioning is an effective method of gathering the perceptions of survivors. CONCLUSIONS: Investigating survivors' impressions of building bombing hazards is critical to capture injury exposures, behavior patterns, and decision-making processes during actual events, and to identify interventions that will be supported by survivors. PMID- 19557965 TI - Risk and protective factors that impact survival in emergencies--the time has come for an upgrade in preventive emergency population behavior. PMID- 19557967 TI - Hospital preparedness in earthquake zones: a must. PMID- 19557966 TI - Earthquake related injuries: assessment of 854 victims of the 2003 Bam disaster transported to tertiary referral hospitals. AB - BACKGROUND: In December 2003, the residents of Bam, Iran experienced an earthquake that measured 6.6 on the Richter scale and destroyed >90% of the city. After the assessment and initial treatment of injuries at national and international field hospitals, a considerable number of victims (approximately 12,000) were transferred to tertiary referral hospitals around the country. OBJECTIVE: This report evaluated the injuries of 854 victims transferred to 12 referral hospitals in Tehran. METHODS: The demographic data, injury patterns, injury severity score (ISS), diagnosis, treatment, and outcome data of 854 Bam earthquake victims were assessed. RESULTS: There were 467 (54.7%) males and 387 (45.3%) females. The mean age of the patients was 29.0 years. Transportation by aircraft was the most common method used for evacuation, which was used to evacuate 555 patients (65%). Fifty-four percent of the victims required initial medical aid at field hospitals before transportation to Tehran. There were 1,322 patients with injuries, of which, fractures of the lower extremities were the most common (331; 25%). Limb fixation was the most commonly performed primary procedure in emergency wards (389 cases, 39.9%). The mean value +/- SD for ISS was 6.7 +/- 5.2. Orthopedic operations were the most frequent surgical procedures performed (195/260 operations, 75%) and the overall mortality rate was 1.6% (n = 14). CONCLUSIONS: Along with the crucial importance of aid provided by national and international field hospitals in disasters, suitable triage of casualties and preparedness of tertiary referral centers in unaffected regions also play an important role in providing medical care to disaster victims. During these situations, the number of victims cannot be predicted accurately, and sufficient medical care, particularly for orthopedic problems, can be provided by referral centers. PMID- 19557968 TI - Injury patterns and levels of care at a marathon. AB - INTRODUCTION: Marathons pose many challenges to event planners. The medical services needed at such events have not received extensive coverage in the literature. OBJECTIVE: The objective of this study was to document injury patterns and medical usage at a category III mass gathering (a marathon), with the goal of helping event planners organize medical resources for large public gatherings. METHODS: Prospectively obtained medical care reports from the five first-aid stations set up along the marathon route were reviewed. Primary and secondary reasons for seeking medical care were categorized. Weather data were obtained, and ambient temperature was recorded. RESULTS: The numbers of finishers were as follows: 4,837 in the marathon (3,099 males, 1,738 females), 814 in the 5K race (362 males, 452 females), and 393 teams in the four-person relay (1,572). Two hundred fifty-one runners sought medical care. The day's temperatures ranged from 39 to 73 degrees F (mean, 56 degrees F). The primary reasons for seeking medical were medication request (26%), musculoskeletal injuries (18%), dehydration (14%), and dermal injuries (11%). Secondary reasons were musculoskeletal injuries (34%), dizziness (19%), dermal injuries (11%), and headaches (9%). Treatment times ranged from 3 to 25.5 minutes and lengthened as the day progressed. Two-thirds of those who sought medical care did so at the end of the race. The majority of runners who sought medical attention had not run a marathon before. CONCLUSIONS: Marathon planners should allocate medical resources in favor of the halfway point and the final first-aid station. Resources and medical staff should be moved from the earlier tents to further augment the later first-aid stations before the majority of racers reach the middle- and later distance stations. PMID- 19557969 TI - Paramedic identification of electrocardiograph J-point and ST-segments. AB - INTRODUCTION: Correct identification of the J-Point and ST-segment on an electrocardiograph (ECG) is an important clinical skill for paramedics working in acute healthcare settings. The skill of ECG analysis and interpretation is known to be challenging to learn and often is a difficult concept to teach. OBJECTIVES: The objective of the study was to determine if undergraduate paramedic students could accurately identify ECG ST-segment elevation and J-Point location. METHODS: A convenience sample of undergraduate paramedic students (n = 148) was provided with four enlarged ECGs (ECG1-4) that illustrated different levels, patterns, and characteristics of ST-segment elevation. Participants were asked to identify whether ST-elevation was present, and if so, height in millimeters (mm) and the correct location of the J-Point. RESULTS: There were significant variations in students' accuracy with both J-Point and ST-segment determination. Eleven (10%) students correctly identified the ST-segment being present in all ECGs. Also, ECG 2 reflected 6 mm of ST-elevation; however, only one student correctly identified this. Overall the students were 0.55 mm (95% CI = 0.29-0.81 mm, range = -6.5-5.8 mm) from the J-point on the horizontal and -0.18 mm (95% CI = -0.31-0.04 mm, range = -2.8-2.3 mm) on the vertical axis. CONCLUSIONS: Undergraduate paramedic students recognize ST-segment elevation. However, inaccuracies occurred with measurements of ST-segment and precise location of J-Points. Errors in ECG analysis may reflect weaknesses in teaching this skill. Consideration should be given to the design of an educational program that can reliably improve performance of this skill. PMID- 19557970 TI - Acquiring and maintaining competence in the application of extrication cervical collars by a group of first responders. AB - INTRODUCTION: Research on skill acquisition and retention in the prehospital setting has focused primarily on resuscitation and defibrillation. Investigation into other first aid skills is required in order to validate practices and support training regimes. No studies have investigated competency using an extrication cervical collar for cervical spine immobilization. OBJECTIVE: This study was conducted to confirm that a group of first responders could acquire and maintain competency in the application of an extrication cervical collar over a 12-month period. METHODS: Participants attended a standardized training session that addressed the theory of application of an extrication cervical collar followed by hands-on practice. The training was presented by the same instructor and covered the nine key elements necessary in order to be deemed competent in extraction cervical collar application. Following the practical session, the competency of the participants was assessed. Participants were requested not to practice the skill during the 12-month period. Following the 12-month period, their skills were re-assessed by the same assessor. RESULTS: Of the 64 subjects who participated in the study, 100% were competent after the initial first assessment. Forty-one participants (64%) were available for the second assessment (12 months later); of these, 25 (61%) maintained competence. CONCLUSIONS: Although the sample size was small, this research demonstrates that first responders are able to acquire competence in applying an extrication cervical collar. However, skill retention in the absence of usage or re-training is poor. Larger studies should be conducted to validate these results. In addition, there is a need for research on the clinical practice and outcomes associated with spinal immobilization in the prehospital setting. PMID- 19557971 TI - Stress, work overload, burnout, and satisfaction among paramedics in Israel. AB - INTRODUCTION: The number of paramedics in Israel is increasing. Despite this growth and important role, the emergency medical organizations lack information about the characteristics of their work. OBJECTIVE: The objective of this study was to examine the characteristics of the paramedics' work, the quality of their working lives, the factors that keep them in the profession, or conversely, draw them away from it. METHODS: Cross-sectional study conducted through telephone interviews of a random sample of 50% of the graduates of paramedic courses in Israel (excluding conscripted soldiers). RESULTS: The factors that attract paramedics to the profession have much to do with the essence of the job-rescuing and saving-and a love of what it involves, as well as interest and variety. Pressures at work result from having to cope with a lack of administrative support, paperwork, long hours, imbalance between work and family life, and salary. They do not come from having to cope with responsibility, the pressure of working under uncertain conditions, and the sudden transition from calm situations to emergencies. Dissatisfaction at work is caused by burnout, work overload, and poor health. Physical and mental health that impedes their ability to work is related to a sense of burnout and the intention to change professions. CONCLUSIONS: The findings about the relationships between health, job satisfaction, and burnout, coupled with the fact that within a decade, half of the currently employed paramedics will reach an age at which it is hard for them to perform their job, lead to the conclusion that there is a need to reconsider the optimum length of service in the profession. There also is a need to form organizational arrangements to change the work procedures of aging paramedics. PMID- 19557972 TI - Seatbelt use in Qatar in association with severe injuries and death in the prehospital setting. AB - INTRODUCTION: Road traffic crashes (RTCs) are common in Qatar, and are now considered the third leading cause of mortality. In this study, the safety devices used by the Qatari public at the time of RTCs were assessed and the association between seatbelt use by vehicle occupants involved in RTCs and severe injury/death in the prehospital setting was determined. METHODS: This study was a retrospective case-control investigation. A Hamad Medical Corporation Emergency Medical Services (EMS) database of RTCs occurring from January 2006 to April 2007 was utilized for this study, providing a total of 5,267 patient records (83.5% male, 16.5% female, median age = 28 years). Patient demographics, crash characteristics, prehospital assessments, and interventions were identified, and use of safety devices was determined. Univariate analysis including chi-square, Student's t-test, and analysis of variance (ANOVA) was performed as appropriate. "Case" patients are defined as those who had specific, critical prehospital assessments, or who received advanced cardio-respiratory life support measures in the field. Logistic regression modeling was used to predict the probability of a case being unbelted, controlling for confounders. RESULTS: Seatbelt use in Qatar was low: 33.9% of males and 32.6% of females wore seatbelts at the time of the RTC. Victims involved in a vehicle rollover crash were less likely to be belted than were those involved in a non-rollover incident (26.2% belted vs. 37.8%; OR = 0.59; 95% CI = 0.50-0.68). Case patients-those with defined critical assessment findings or resuscitation in the field-and control patients were similar in age (30 years vs. 28 years median). Case patients were disproportionately male (89.1% vs. 83.2%; OR = 1.65; 95% CI = 1.01-2.83) and were more likely to be victims of a vehicle rollover crash (44.7% vs. 23.8%; OR = 2.57; 95% CI = 1.84-3.59). Seatbelt use was significantly lower among cases than controls: 19.7% of cases were reported to have worn seatbelts compared to 34.2% of controls (OR = 0.47; 95% CI = 0.31-0.69). This relationship also persisted (OR = 0.51; 95% CI = 0.33-0.76) after controlling for confounders. CONCLUSIONS: Seatbelt use in Qatar is low. Seatbelts are protective: in the prehospital setting unbelted vehicle occupants involved in RTCs were nearly twice as likely to suffer severe injury or death compared to belted patients. Prehospital morbidity and mortality appears to be reduced significantly by the consistent use of seatbelts by the motoring population in Qatar. PMID- 19557973 TI - Needle thoracostomy by non-medical law enforcement personnel: preliminary data on knowledge retention. AB - INTRODUCTION: Tension pneumothorax is the second leading cause of preventable combat death. Although relatively simple, the management of tension pneumothorax is considered an advanced life support skill set. The purpose of this study was to assess the ability of non-medical law enforcement personnel to learn this skill set and to determine long-term knowledge and skill retention. METHODS: After completing a pre-intervention questionnaire, a total of 22 tactical team operators completed a 90-minute-long training session in recognition and management of tension pneumothorax. Post-intervention testing was performed immediately post-training, and at one- and six-months post-training. RESULTS: Initial training resulted in a significant increase in knowledge (pre: 1.3 +/- 1.35, max score 7; post: 6.8 +/- 0.62, p < 0.0001). Knowledge retention persisted at one- and six-months post-training, without significant decrement. CONCLUSIONS: Non-medical law enforcement personnel are capable of learning needle decompression, and retain this knowledge without significant deterioration for at least six months. PMID- 19557974 TI - Near-infrared laser spectroscopy as a screening tool for detecting hematoma in patients with head trauma. AB - INTRODUCTION: Among imaging techniques, computed tomography (CT) is a reliable method for detecting intracranial hematomas in patients with head trauma, but it is not generally available in special circumstances like prehospital situations and harsh conditions such as those following an earthquake. OBJECTIVE: The objective of this study is to determine if near-infrared spectroscopy (NIRS) is useful for performing CT scans on patients with closed head trauma that present to medical centers that do not have the ability to perform a CT scan or in prehospital or harsh situations. Near-infrared spectroscopy and CT scan were compared according to sensitivity and specificity. METHODS: This was an observational, prospective study. One hundred forty-eight patients admitted to Rasool Akram General Hospital in Tehran, Iran with head injuries during a one month period were studied using NIRS and CT. The observational, prospective study was conducted and sensitivity, specificity, positive and negative predictive values of NIRS were calculated. Chi-square and Kappa analysis was performed, and a p-value <0.05 was considered significant. RESULTS: According to the CT scan findings, 54 (36.5%) of the patients developed intracranial hematoma. The NIRS examination showed that 69 patients (46.6%) might have intracranial hematoma. The number of true negatives was 73 and the number of false negatives was six patients. The sensitivity and specificity of NIRS examination was 88.9% and 77.7%, respectively. CONCLUSIONS: This study speculates that NIRS may be a useful screening tool to detect intracranial hematoma. This capability could be useful in special situations like in a deprived area, medical centers without CT scan capabilities, prehospital situations, and in harsh conditions like those after an earthquake or other disasters that causes increased numbers of victims with closed head trauma. PMID- 19557975 TI - Exercise stress testing in asymptomatic severe aortic stenosis. AB - Aortic valve replacement is the accepted therapy for patients with severe symptomatic aortic stenosis (AS). However, the timing of surgery in asymptomatic patients is less well defined, as some -- but not all -- asymptomatic patients are at increased risk. Exercise stress testing is an attractive means to assess risk in such patients, as it is readily available, standardized, physiological, and presents a controlled environment in which to assess whether patients truly are asymptomatic. This review summarizes existing literature and current guideline recommendations addressing exercise testing in asymptomatic patients with AS, and provides a recommendation for its use in a subset of patients. PMID- 19557976 TI - Valve replacement in octogenarians: arguments for an earlier surgical intervention. AB - BACKGROUND AND AIM OF THE STUDY: In octogenarians with symptomatic aortic valve stenosis (AS), aortic valve replacement (AVR) is frequently not performed in due time, because the prognostic benefit is underestimated, while perioperative morbidity and mortality are overestimated. The severely impaired prognosis and quality of life after myocardial decompensation then urges AVR with a significantly increased perioperative risk. METHODS: Between 2003 and 2006, all octogenarians with isolated symptomatic AS (indexed aortic valve opening area <0.5 cm2/m2) referred to the authors' unit were prospectively included in the survey. Among the 83 patients enrolled (51 women, 32 men; mean age 84 +/- 5.1 years), 38 patients (26 women, 12 men; mean age 84 +/- 2.3 years) had signs of chronic myocardial decompensation (dilated left ventricle and/or reduced left ventricular function; left ventricular ejection fraction (LVEF) 43 +/- 18% (range: 25-53%). These patients comprised group A. All other patients (group B) had normal left ventricular dimensions, a normal LVEF (>55%), and no clinical episodes of myocardial decompensation. All patients underwent AVR, while 23 (28%) underwent simultaneous coronary revascularization. RESULTS: In group A, the 30 day mortality rate was 5.3% (n = 2). Octogenarians without chronic myocardial decompensation had a lower 30-day mortality (1/45; 2.2%). The incidences of major postoperative complications (reversible acute renal failure, stroke, mechanical circulatory support) were significantly higher in group A (26.3% versus 8.9%, p < 0.05). During late follow up (mean 24.2 +/- 12.8 months), another four patients in group A (11.1%) and five in group B (11.4%) died. Octogenarians in group B had a significantly (p < 0.01) more favorable cumulative survival rate (87% versus 78% after 24 months; 81% versus 68% after 46 months). CONCLUSION: AVR can be performed in octogenarians with a low mortality and morbidity, but should not be postponed. The decision to perform for AVR may take into consideration any life limiting comorbidities, but should be made independent of the patient's age. PMID- 19557977 TI - Live longer and better without prosthesis-patient mismatch. PMID- 19557978 TI - Patient-prosthesis mismatch in elderly patients undergoing aortic valve replacement: impact on quality of life and survival. AB - BACKGROUND AND AIM OF THE STUDY: Since the introduction of its theoretical basis, patient-prosthesis mismatch (PPM) following aortic valve replacement (AVR) has been the subject of much debate. The study aim was to compare, by a propensity score adjustment, the survival and quality of life in elderly patients with PPM, to those of a population without mismatch. The analysis was focused on elderly patients, as their high prevalence of calcific aortic stenosis may increase the probability to receive a small-sized aortic prosthesis, and consequently to experience postoperative PPM. METHODS: A total of 163 patients aged > or = 75 years who underwent AVR was analyzed. The median logistic euroSCORE was 7.1%. PPM was considered to be present if the anticipated indexed effective orifice area (IEOA) was < or = 0.85 cm2/m2. The median follow up period was 37.4 months. The patients' quality of life was evaluated using the Short Form 12 (SF-12) Health Survey test. RESULTS: PPM was present in 43% of the patients. In multivariable analysis, patients with PPM were more often female, more often operated on for aortic degenerative calcification, had a larger body surface area, and more often received a bioprosthesis than those without mismatch. The survival analysis did not highlight any significant difference between the two groups. According to a multivariable analysis, the SF-12 physical component score of PPM patients was significantly inferior to that in patients without mismatch (p = 0.001). CONCLUSION: The study results suggest that moderate PPM does not have a negative impact on mid-term mortality in elderly patients after AVR. However, PPM was associated with a reduced quality of life in this elderly population. PMID- 19557979 TI - Risk of reoperation for aortic bioprosthesis dysfunction. AB - BACKGROUND AND AIM OF THE STUDY: The risk of reoperative valve replacement for failed aortic bioprosthesis may be overestimated, this being a dominant factor when selecting an initial prosthesis to be implanted in patients aged <70 years. The study aims were, first, to analyze the mortality and morbidity of redo aortic bioprosthesis replacement in the current era, and second, to identify preoperative risk factors and evaluate the EuroSCORE. METHODS: A total of 156 consecutive patients (111 men, 45 women; mean age 60.9 years; range: 23-87 years) who underwent reoperation for failed aortic bioprosthesis between 1990 and 2006 was reviewed in this retrospective, single-center study. Surgery was undertaken due to bioprosthesis degeneration (82.7%), bacterial prosthetic endocarditis (14.1%), paravalvular leak (1.3%) and other causes (1.9%). Emergency procedures were performed in 9% of patients. Associated procedures were coronary artery bypass grafting in 7.7% of patients, ascending aortic graft in 7%, and complete aortic root replacement in 6.4%. The predictive mortality was 8% according to the Additive EuroSCORE, and 15% according to the Logistic EuroSCORE. RESULTS: Overall, the operative mortality was 3.8% (n = 6), and postoperative morbidity was low. The only multivariable predictor was emergency surgery (OR = 15.22, 95% CI = 1.68-86.43; p = 0.02). A mortality trend was associated with atrial fibrillation and NYHA class III/IV, but this was not statistically significant (p = 0.09 and p = 0.06, respectively). Associated procedures were not significant risk factors for mortality. CONCLUSION: Reoperation for aortic bioprosthesis dysfunction can be performed with a low risk of mortality. It appears that this risk is overestimated by the EuroSCORE. Those patients who wish to avoid postoperative anticoagulant therapy may choose to receive this type of valve, even if reoperation is foreseeable. PMID- 19557980 TI - Parathyroid hormone gene variant and calcific aortic stenosis. AB - BACKGROUND AND AIM OF THE STUDY: Calcific aortic stenosis (AS) is the most frequently acquired valvular disease of the elderly in the Western world. A genetic background for AS has been proposed. The deposition of calcium hydroxyapatite is the key problem of valve calcification; vitamin D and parathyroid hormone are major factors in calcium homeostasis. The vitamin D receptor (VDR) gene and parathyroid hormone (PTH) gene variants were selected as candidate genes. METHODS: A total of 538 patients with severe calcific AS (identified echocardiographically) were characterized by left heart catheterization. A group of 536 patients in whom heart disease had been excluded by left heart catheterization served as a control population. The cardiovascular risk profile was assessed, and three gene variants were analyzed, namely VDR rs1544410, VDR rs1073810, and PTH rs6254. RESULTS: Patients with AS were found to have a higher prevalence of the PTH AA genotype (108 +/- 20.1% versus 71 +/- 13.2%; p = 0.007), while the VDR gene revealed a marginal, but statistically non significant, association. The age and risk profile was similar in both groups. CONCLUSION: To date, the association of the PTH gene variant has been the only positive association studied in patients with AS in a large population. Hence, the polymorphism is within an intron; the molecular mechanisms of altered gene expression should undergo further investigation. PMID- 19557981 TI - Predictors of the outcome of thrombolytic therapy in prosthetic mitral valve thrombosis: a study of 62 events. AB - BACKGROUND AND AIM OF THE STUDY: Treatment strategies and guidelines in the management of prosthetic valve thrombosis (PVT) remain controversial. The study aim was to assess the success and complication rate of thrombolytic therapy (TT) by comparing thrombus size, severity of symptoms, type of prosthetic valve, and time since valve implantation. METHODS: Between April 1993 and December 2006, TT was administered for 62 thrombotic events in 55 patients with mitral PVT. Thrombus size was measured by planimetry, and its location and mobility assessed with transesophageal echocardiography (TEE). Continuous thrombolytic treatment in obstructive PVT (OPVT) was monitored using gradient measurement by transthoracic echocardiography every 2-5 h, and by visual assessment with TEE every 24 h. RESULTS: All patients with non-obstructive PVT (NOPVT) were in NYHA class I or II. Among 52 patients with OPVT, eight were in NYHA class I or II, and 44 in NYHA class III-IV (p < 0.0001). The thrombus area (measured with TEE) before thrombolysis was < 0.8 cm2 in 24 cases, > or = 0.8 cm2 in 21 cases, and could not be measured in 17 events. Thrombolysis was successful in 45 events (73%) including all NOPVT. In 13 events (21%) the transvalvular gradient decreased but was not normalized. Thrombolysis failed in four events (6%). Complications of thrombolysis were present in 11 events (18%), and four patients died. There was no significant difference in the outcome of TT regarding the type of artificial valve, NYHA class or thrombus size. In cases of successful thrombolysis, the time since surgery was significantly shorter than in cases of partially successful or failed thrombolysis. CONCLUSIONS: Based on previous data and the present findings, thrombolysis can be considered as first-line treatment in all patients with PVT, independent of the valve type, NYHA functional class, and thrombus size. PMID- 19557982 TI - Mitral regurgitation reduces systemic coagulation activity in rheumatic heart disease: using fibrin D-dimer as a biomarker of thrombogenesis. PMID- 19557983 TI - Mitral regurgitation reduces systemic coagulation activity in patients with rheumatic heart disease. AB - BACKGROUND AND AIM OF THE STUDY: Severe mitral regurgitation (MR) reduces left atrial thrombus formation in patients with mitral stenosis (MS) and atrial fibrillation (AF). Plasma D-dimer levels represent a biochemical marker for fibrinolytic activity in prothrombotic states. The prothrombotic burden in patients with mitral valve disease and/or AF was assessed using plasma D-dimer levels. METHODS: The study population included 89 patients with mitral valve disease, 21 with AF but normal valves, and 15 healthy controls. The mitral valve group was subdivided into patients with MS (n = 27), severe MR (n = 26), and MS with concomitant with severe MR (MS/MR; n = 36). These subgroups were further subdivided according to the atrial rhythm (sinus rhythm (SR)+AF). RESULTS: The mean left atrial size was increased in all groups with cardiac disease. D-dimer levels were highest in the MS+AF subgroup (527 +/- 134 microg/l). Patients with MS+AF, MS+SR, and nonvalvular AF had significantly higher D-dimer levels than controls (p < 0.01, by ANOVA). Patients with MR had normal levels of D-dimer. The atrial rhythm did not influence D-dimer levels in the MS/MR subgroup, or in the pure MR subgroup (p = NS). CONCLUSION: Plasma D-dimer levels correlated with the embolic risk in mitral valve disease and non-valvular AF. The highest levels were found in patients with MS+AF and non-valvular AF. Severe MR decreased the D-dimer levels in MS and/or AF to control levels. PMID- 19557984 TI - Pure annular dilation as a cause of mitral regurgitation: a clinically distinct entity of female heart disease. AB - BACKGROUND AND AIM OF THE STUDY: Pure annular dilation (PAD) is a recognized etiology of mitral regurgitation, yet few data exist to define the prognostic profile of this disorder relative to other etiologies, such as ischemia or myxomatous prolapse. METHODS: A total of 535 patients undergoing mitral repair at two institutions between 1993 and 2002 was retrospectively reviewed. PAD was defined as requiring only ring annuloplasty +/- cleft repair, without evidence of prolapse, regional wall motion abnormality, or infarction. RESULTS: PAD was identified in 74 patients, while alternative etiologies were myxomatous prolapse (n = 290), ischemia (n = 141), and 'other' (n = 30). PAD patients were more often female (78%) than male (38%) (p < 0.001), more often hypertensive (37% versus 26%; p = 0.003), and had a left ventricular ejection fraction (LVEF) that was lower (0.41 +/- 0.12) than those in patients with prolapse (0.51 +/- 0.11; p < 0.01) but similar to values in ischemic patients (0.38 +/- 0.10). The valve size was smaller for PAD versus prolapse (ring size 24-26 mm in 71% versus 12%; p < 0.001). The unadjusted PAD prognosis was intermediate, with five-year survival being 70 +/- 8%, compared to 87 +/- 3% for prolapse and 56 +/- 5% for ischemia (p < 0.01). Survival adjusted for differences in baseline characteristics was not different among the three groups (p > 0.10). CONCLUSION: PAD is a clinically distinct etiology of mitral regurgitation associated with female gender, small valve size, a lower LVEF, and hypertension. Early, more aggressive hypertension control might improve or minimize the consequences of this predominantly female cardiac disorder. PMID- 19557985 TI - Gaping cleft or commissure--an under-rated cause of residual mitral insufficiency following valve repair: case reports. AB - Naturally occurring clefts in the posterior leaflet of the mitral valve and/or the mitral commissures themselves may become the foci of residual mitral regurgitation when distorted by an adjacent rigid suture line. Herein are reported the details of three cases in which cleft/commissure closure resolved such leaks. The anatomical substrate which predisposed to this problem is also discussed. PMID- 19557986 TI - Ultrastructure of porcine mitral valve chordae tendineae. AB - BACKGROUND AND AIM OF THE STUDY: The chordae tendineae, which form an important component of the mitral valve apparatus, experience continuous cyclic loading and are thus well-adapted for effectively storing and dissipating energy. An understanding of their microstructure would be expected to shed light on the mechanism of their remarkable durability. METHODS: In these studies, porcine mitral valve chordae from freshly slaughtered pigs were used. Histological samples of Picrosirius Red-stained and Movat's pentachrome-stained chordae were examined with optical microscopy and laser scanning confocal microscopy. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to study the ultrastructure at high magnification. RESULTS: Both, optical microscopy and SEM revealed that the waviness of collagen fibers was uniform across the thickness of the chordae, with no straight fibers in the core. Wavy fibers and fiber bundles were found to be in skewed-register, rather than transverse. Collagen fiber bundles were found to undulate in a three-dimensional path, rather than the planar waveform, as reported previously. TEM showed that different types of chordae had different fibril configurations. Marginal chordae had smaller diameters but a higher fibril density than did basal and strut chordae. CONCLUSION: The configuration of collagen fibrils in the mitral valve chordae is more complex than initially thought, and different chordae have morphologies that are likely specific to their mechanical role in the mitral apparatus. These findings provide insight into possible improvements for chordal repair surgery, and form a structural basis for accurate computational modeling. PMID- 19557987 TI - Mitral web--a new concept for mitral valve repair: improved engineering design and in-vitro studies. AB - BACKGROUND AND AIM OF THE STUDY: A new mitral valve repair concept to treat mitral regurgitation (MR) due to valve prolapse was recently proposed. In this study, an improved design of this concept is presented, and the results of preliminary hemodynamic studies conducted in an in-vitro prolapse model are reported. METHODS: The new repair approach is based on using a web/net attached to a standard annuloplasty ring spanning the annulus of the mitral valve. Experiments were conducted in a left ventricular simulator, using native porcine mitral valves. Severe MR was created by transecting the marginal chordae to induce P2 prolapse, and also by displacing the papillary muscles basally to induce bileaflet prolapse. RESULTS: Implantation of the mitral web prevented leaflet prolapse and restored leaflet coaptation with trace mitral insufficiency, in both posterior and bileaflet prolapse. In posterior leaflet prolapse, implantation of the mitral web reduced the regurgitation volume from 10.43 +/- 3.76 ml to 2.13 +/- 1.83 ml per beat (p < 0.05). No visual damage was observed on the mitral valve leaflets after 4 h of continuous operation. CONCLUSION: Based on the study results, the mitral web may represent a feasible option to repair different types of mitral valve prolapse. The mitral web may also significantly simplify mitral valve repair for complex lesions. PMID- 19557988 TI - Management of submitral anomalies during complex mitral valve repair: a discussion of techniques. PMID- 19557989 TI - Guidelines for the prevention of infective endocarditis revised: a critical appraisal of the AHA recommendations and call for an individual approach. PMID- 19557990 TI - Results of annular reconstruction with a pericardial patch in active infective endocarditis. AB - BACKGROUND AND AIM OF THE STUDY: Annular reconstruction in active infective endocarditis (IE) is technically difficult, and results in a high mortality and morbidity. The study aim was to determine the midterm results of annular reconstruction with a pericardial patch in active IE. METHODS: A total of 57 operations was performed in 56 patients (38 men, 18 women; mean age 48.3 +/- 16.9 years) with active IE. Twenty-five cases (44%) were included in whom the preoperative NYHA class was III or IV. RESULTS: Bovine pericardium was used in 52 cases, autologous pericardium in three, and bovine + autologous pericardium in two. The aortic annulus was reconstructed in 18 cases, combined with aortomitral continuity in 13 cases, and both aortic and mitral annulus were combined with aortomitral continuity in three cases. The mitral annulus was reconstructed in 21 cases, and the complete cardiac skeleton was reconstructed in one case. There were three operative deaths. Postoperative complications included reexploration due to bleeding in two cases, mediastinitis in one case, complete atrioventricular block in five cases, and cerebral hemorrhage in six cases. The follow up was 93% complete (52/56); the mean duration of follow up was 45.1 +/- 32.6 months (range: 2-138 months). There were two late deaths, at three and eight months postoperatively. Endocarditis recurred five times in four patients. Re-do surgery was performed in four cases due to endocarditis recurrence in three patients at two, three, and 29 months after surgery, respectively, and to a pseudoaneurysm in one patient at one month postoperatively. The mean survival at two years was 91 +/- 3.9%; the two-year event-free survival was 82 +/- 5.4%. CONCLUSION: Annular reconstruction with a pericardial patch in active IE can be performed safely, and showed good durability at the mid-term follow up examination. PMID- 19557991 TI - An alternative surgical approach to infective mitral endocarditis. PMID- 19557992 TI - A postoperative Gerbode defect in aortic prosthesis endocarditis with non-typhoid Salmonella. AB - The Gerbode defect is a congenital shunt from the left ventricle to the right atrium. The type I defect (2) results in a direct shunt through a portion of the membranous septum, while a type II (indirect) defect occurs if the membranous septal defect lies below the attachment of the septal leaflet of the tricuspid valve. The shunt is directed towards the right atrium through a cleft or perforations of the septal leaflet. Acquired Gerbode defects have been identified in endocarditis, after mitral or aortic valve surgery, or may be post-traumatic. The case is presented of a 69-year-old woman with a postoperative Gerbode defect in association with aortic prosthetic endocarditis caused by non-typhoid Salmonella. PMID- 19557993 TI - High prevalence of rheumatic heart disease by clinical and echocardiographic screening among children in Fiji. AB - BACKGROUND AND AIM OF THE STUDY: Rheumatic heart disease (RHD) is an important cause of morbidity and mortality in young people in developing countries. Many cases of RHD are first detected when they progress to cardiac failure. Screening for RHD represents a means of detecting cases early so that preventative measures to halt the disease progression can be put into place. METHODS: A cross-sectional screening survey of RHD in 3,462 children aged 5 to 15 years in Fiji was performed in 2006. A three-stage screening method was used: stage 1 involved auscultatory screening; stage 2 was a limited echocardiography of children identified as having a suspicious murmur in stage 1; and stage 3 involved a full echocardiography of children identified as having pathology in stage 2. RESULTS: Among the 3,462 children screened, 359 (10.4%) had a significant murmur; subsequent echocardiography was performed on 331 of these children, with RHD being detected in 29 cases. The prevalence of definite RHD was 4.1 per 1,000 (95% CI 2.2-6.8), and the overall prevalence (definite or probable RHD) was 8.4 cases per 1,000 (95% CI 5.6-12). CONCLUSION: The study results suggest that there is a significant burden of undetected RHD in Fiji. The three-stage approach described here represents a practical means of screening for clinical RHD in developing countries, although it does not allow detection of the subclinical disease. PMID- 19557994 TI - The incidence of congenital bicuspid or bileaflet and quadricuspid or quadrileaflet arterial valves in 3,861 donor hearts in the European Homograft Bank. AB - BACKGROUND AND AIM OF THE STUDY: The incidence of bicuspidity and quadricuspidity of arterial valves in donor hearts has been assessed in the European Homograft Bank. METHODS: Morphologically unacceptable donor valves for clinical use were assessed for cusp or leaflet malformation, identifying the different valve types. The state of the adjacent valve was also evaluated. Histological evaluations of the corresponding conduits were performed, in order to assess the incidence of media degeneration. RESULTS: Among 3,861 donor hearts, 39 (1.0%) showed congenital cusp or leaflet malformation of one or both arterial valves, 28 (0.7%) showed bicuspid or bileaflet aortic valves (BCAV), four (0.1%) showed bicuspid or bileaflet pulmonary valves (BCPV), and eight (0.2%) showed quadricuspid or quadrileaflet pulmonary valves (QCPV). Only in one case (0.03%) were both arterial valves bicuspid or bileaflet. Among the bicuspid arterial valves, 27 (84%) were isolated aortic valves, three (9%) were isolated pulmonary valves, and in one case (3%) both the aortic and pulmonary valves were bicuspid or bileaflet. CONCLUSION: The most frequently identified congenital malformation of the arterial valves was aortic bicuspidity, followed by pulmonary quadricuspidity and bicuspidity. If asymptomatic, these were detected during routine echocardiography, or at autopsy. These malformations might be genetically determined, as they are detected more often among different members of the same families, and in those persons of the same gender or blood group. These valves have a predisposition to accelerated calcification, stenosis, endocarditis and dissection of the ascending aorta. Neither quadricuspid/quadrileaflet nor unicuspid/monoleaflet aortic valves were detected in this study. PMID- 19557995 TI - Traumatic aortic valve rupture after a high-speed motor-vehicle accident: case report. AB - The case is reported of a 32-year-old male with an aortic valve injury received in a high-speed traffic accident. Several non-cardiac concomitant injuries were present, none of them life-threatening. Cardiac surgery was performed on day 2 due to a grade III aortic valve insufficiency. The presence of a large tear of the non-coronary cusp, and several concomitant injuries, led to valve replacement being considered the most optimal treatment for this patient. PMID- 19557996 TI - Supravalvular aortic membrane with severe aortic regurgitation: an unusual presentation in adults. AB - Aortic stenosis due to supravalvular membrane usually presents in children. It may be associated with fusion of the left coronary leaflet and the supravalvular membrane, causing obstruction of the left coronary ostium, and resulting in myocardial ischemia. Despite the immobilization of the left coronary leaflet, these patients present in childhood with aortic stenosis and not regurgitation, with or without accompanying myocardial ischemia. The case is described of an adult patient with supravalvular aortic membrane presenting with severe aortic regurgitation and myocardial infarction due to fusion of the left coronary leaflet with the supravalvular membrane. PMID- 19557997 TI - Idiopathic mitral valve disease in a patient presenting with Axenfeld-Rieger syndrome. AB - A 33-year-old, previously healthy male presented with respiratory distress and underwent intubation. A physical examination revealed a holosystolic murmur and pupillary abnormalities. Echocardiography revealed a flail anterior mitral valve leaflet with ruptured chordae and severe mitral regurgitation. The patient underwent urgent mitral valve replacement and tolerated the procedure well. The mitral valve leaflet was myxomatous and calcified -- an unusual find in such a patient. An ophthalmology consultation was obtained and the patient diagnosed with Axenfeld-Rieger syndrome, a disorder of the anterior ocular chamber that has been associated with cardiac malformations. The present case report adds to the body of literature which suggests a correlation between Axenfeld-Rieger syndrome and valvular abnormalities. Hence, it is believed prudent that patients with Axenfeld-Rieger syndrome should undergo echocardiographic screenings for valvular abnormalities. PMID- 19557998 TI - Left ventricular rupture after mitral valve replacement: risk factor analysis and outcome of resuscitation. PMID- 19557999 TI - Mitral acute bacterial endocarditis caused by HACEK microorganisms. PMID- 19558000 TI - Mango, Mangifera indica. PMID- 19558001 TI - T-cell epitopes of aeroallergens. AB - OBJECTIVE: To describe the current knowledge of the T-cell epitopes of common aeroallergens, how they were discovered, and implications for future therapy. DATA SOURCES: PubMed search of English-language articles without date limits pertaining to T-cell epitopes of aeroallergens included on a standard skin test panel. STUDY SELECTION: A total of 127 articles were screened based on the results of the PubMed search and cross-indexed as needed. The highest quality and most clinically relevant articles were included for discussion. RESULTS: Of the 47 allergen extracts included on the standard skin test panel at our instittition, T-cell epitopes have been described for 13. Immunodominant epitopes have been used for peptide immunotherapy trials. CONCLUSIONS: T-cell epitopes have been characterized for a minority of common aeroallergens. However, knowledge is rapidly expanding and can lay the groundwork for therapies that specifically target T cells. PMID- 19558003 TI - The role of physical activity and body mass index in the health care use of adults with asthma. AB - BACKGROUND: Health care use in patients with asthma is affected by many factors, including sex and ethnicity. The role of physical activity (PA) and body mass index (BMI) (calculated as weight in kilograms divided by height in meters squared) in this relationship is unknown. OBJECTIVE: To determine the role of PA and BMI in the health care use of patients with asthma. METHODS: A sample of adults with asthma (n=6,835) and without asthma (n=78,051) from cycle 3.1 of the Canadian Community Health Survey was identified. Health care use was self reported as overnight hospital stays (yes or no), length of overnight hospital stay (<4 or > or =4 nights), and physician consultations (<3 or > or =3). Self reported physical activities were used to derive total energy expenditure and to classify participants as active (>3.0 kcal/kg of body weight per day), moderately active (1.5-3.0 kcal/kg of body weight per day), and inactive (<1.5 kcal/kg of body weight per day). The BMI was categorized as normal weight (18.5-24.9), overweight (25.0-29.9), and obese (30.0-59.9). RESULTS: Adjusted logistic regression models revealed that patients with asthma were more likely to have an overnight hospital stay (odds ratio [OR], 2.25; 95% confidence interval [CI], 1.95-2.60), 4 or more overnight hospital stays (OR, 1.48; 95% CI, 1.12-1.96), and 3 or more physician consultations (OR, 2.43; 95% CI, 2.18-2.71) compared with patients without asthma (OR, 1.00). Inactive patients with asthma were more likely to have an overnight hospital stay (OR, 1.68; 95% CI, 1.31-2.16) and 3 or more physician consultations (OR, 1.23; 95% CI, 1.04-1.46) than active patients with asthma (OR, 1.00). Inactive/obese patients with asthma were 2.35 (95% CI, 1.69-3.27) times more likely to have an overnight hospital stay and 2.76 (95% CI, 2.11-3.60) times more likely to have 3 or more physician consultations than active/normal weight patients with asthma (OR, 1.00). CONCLUSIONS: Higher PA levels are associated with lower health care use in individuals with and without asthma. In those with asthma, PA was a more important factor in overnight hospital stays than BMI, whereas both BMI and PA were important determinants of physician consultations. PMID- 19558002 TI - Perception of airflow obstruction in patients hospitalized for acute asthma. AB - BACKGROUND: Little is known about the perception of airflow obstruction in patients hospitalized for acute asthma. OBJECTIVES: To evaluate patient perception of airflow obstruction at hospital discharge and at a 2-week follow-up visit and to determine whether symptom control and/or severity of airflow obstruction identified patients at risk for acute asthma after discharge. METHODS: In a prospective cohort study of inner-city adults hospitalized for acute asthma from April 1, 2001, through October 31, 2002, symptom control (Asthma Control Questionnaire) and airflow obstruction (forced expiratory volume in 1 second [FEV1] percentage predicted) were evaluated at discharge and 2 weeks after discharge. We evaluated perception of airflow obstruction (symptom control vs FEV1 percentage predicted) and perception of change in airflow obstruction (change in symptom control vs percentage change in FEV1) between the 2 visits. Acute asthma after discharge was defined as an emergency department visit or hospitalization for asthma within 90 days of discharge. RESULTS: In fifty-one participants, symptom control was not significantly associated with airflow obstruction at hospital discharge (P = .30), indicating poor perception of airflow obstruction. Among the 41 participants (80.4% of those enrolled) who completed the follow-up visit, change in symptom control was not significantly associated with change in airflow obstruction (P = .20), indicating poor perception of change in airflow obstruction. Greater airflow obstruction at follow-up (P = .02) and a smaller improvement in airflow obstruction (P = .03), but not symptom control, were associated with a higher risk of acute asthma after discharge. CONCLUSIONS: Patients hospitalized for acute asthma have poor perception of airflow obstruction and change in airflow obstruction. Objective measurements of lung function should guide treatment decisions after discharge in this population. PMID- 19558004 TI - Effect of bronchoconstriction on exhaled nitric oxide levels in healthy and asthmatic children. AB - BACKGROUND: Exhaled nitric oxide (eNO) has recently been proposed to be a noninvasive marker of airway inflammation in asthma. OBJECTIVE: To evaluate the effect of bronchoconstriction by means of methacholine inhalation challenge on levels of eNO in children. METHODS: Spirometry, impulse oscillometry, and eNO measurements were performed before and after methacholine inhalation challenge (bronchoconstriction phase) and after beta2-agonist inhalation (bronchodilation phase) in 92 children (62 children with asthma, 13 wheezy children, and 17 healthy children). RESULTS: A significant decrease occurred in the eNO level after methacholine inhalation challenge (P < .01). This decrease did not correlate with the percentage decrease in forced expiratory volume in 1 second or with the change in large airway resistance (R20), but it did correlate with the percentage decline in maximal expiratory flow at 50% vital capacity and with the change in small airway resistance (R5-R20). The eNO decrease lasted for 15 minutes after beta2-agonist inhalation in the group with a high percentage decrease in R5-R20 (>200%). On the other hand, in the group with a low percentage decrease in R5-R20 (< or =200%), eNO recovered to the previous level immediately after beta2-agonist inhalation. CONCLUSIONS: The eNO level significantly decreases after methacholine inhalation challenge. This decrease primarily depends on bronchoconstriction of the small airways. PMID- 19558005 TI - The Urticaria Severity Score: a sensitive questionnaire/index for monitoring response to therapy in patients with chronic urticaria. AB - BACKGROUND: Despite the existence of numerous dermatology-specific quality-of life questionnaires, there exists a need for a well-validated instrument to evaluate and monitor patients with chronic urticaria. OBJECTIVE: To develop and validate a novel chronic urticaria-specific questionnaire, the Urticaria Severity Score (USS). METHODS: The USS was developed with 12 questions and 7 response options per question included in the final questionnaire. The USS was pilot tested on 28 patients with symptomatic chronic urticaria, and a modified version was tested in 80 patients. In this latter group, the USS was compared with the previously validated Dermatology Life Quality Index (DLQI). Patients completed both questionnaires at baseline and at follow-up (median, 2 weeks). Spearman rank correlation tests were used to compare the USS with the DLQI. At a subsequent visit, 9 additional patients completed the USS to evaluate test-retest reliability. RESULTS: There was a positive correlation between the USS and DLQI at baseline (r = 0.64) and follow-up (r = 0.69). The test-retest reliability correlation for the USS was adequate (r = 0.96) (P = .001). The USS was more sensitive in detecting symptom improvement than the DLQI. The mean (SD) "raw score" change from baseline to follow-up for the USS was 9.95 (12.80), whereas the mean (SD) change from baseline to follow-up for the DLQI was 2.25 (4.87). CONCLUSIONS: The USS is a valid and reliable instrument for monitoring urticaria severity. The USS is more sensitive than the DLQI for quantifying urticaria severity and may be moreapplicable in the evaluation of urticaria-specific treatment efficacy. PMID- 19558006 TI - Usefulness of abdominal ultrasonography in the follow-up of patients with hereditary C1-inhibitor deficiency. AB - BACKGROUND: Hereditary angioedema (HAE) is caused by the deficiency of functional C1 inhibitor. Symptoms of this disease include cutaneous angioedema, abdominal pain, and even laryngeal edema. OBJECTIVE: To evaluate the usefulness of abdominal ultrasonography in patients with hereditary C1-inhibitor deficiency in diagnosing acute abdominal edema attacks and possible adverse effects of long term prophylaxis with attenuated androgens. METHODS: Fifty-nine adult patients with HAE regularly observed in our department were included whether they were symptomatic or not and whether they received long-term androgen prophylaxis or not. We evaluated the ultrasonographic findings in the assessments performed routinely or in the moment of an acute abdominal attack. RESULTS: Of the 59 patients, 55 ever had any symptom due to HAE (abdominal location, 78% of the symptomatic patients); 4 patients were asymptomatic. In 11 cases, ultrasonography was performed during acute attacks. Ascites and intestinal wall swelling were found in 7 of these 11 cases and, thus, diagnosis was confirmed. Of the 59 patients, 33 were or had been receiving androgen prophylaxis. Abdominal ultrasonographic assessments were performed routinely in 31 of these patients. Four cases of angiomas, 4 of steatosis, and 1 each of portal hypertension, hepatic cysts, and hepatomegaly were found. Assessments were also performed in 17 patients who did not receive androgen prophylaxis; there were no findings in any of these patients. CONCLUSION: Abdominal ultrasonography has been proved useful as an early tool for diagnosing the adverse effects of therapy and for confirming diagnosis in the case of an acute abdominal attack. PMID- 19558007 TI - Sex-specific differences in allergic sensitization to house dust mites: a meta analysis. AB - BACKGROUND: The prevalence of allergic airway diseases in childhood is higher in boys than in girls but switches toward a female predominance in adolescents and adults. The sex-specific prevalence of allergic sensitization to 1 of the most common allergens worldwide, house dust mite (HDM), has not been examined systematically by age group and species. OBJECTIVE: To systematically review the literature to examine sex-specific differences in the prevalence of allergic sensitization to HDM. METHODS: On the basis of a systematic MEDLINE search for population-based studies published between 1990 and 2007, we conducted meta analyses of male to female ratios for sensitization to 2 different species of HDMs (Dermatophagoides pteronyssinus and Dermatophagoides farinae) separately for children and adults. RESULTS: We included data from 7,822 children and 18,522 adults from 15 studies. The prevalence of sensitization to D pteronyssinus was significantly higher in boys vs girls (male to female ratio, 1.39; 95% confidence interval [CI], 1.13-1.71) and in men vs women (male to female ratio, 1.40; 95% CI, 1.22-1.61). For sensitization to D farinae, the significant male predominance in adults was even more pronounced (male to female ratio, 1.95; 95% CI, 1.37 2.79), but there were no sex-specific differences in children (male to female ratio, 0.90; 95% CI, 0.56-1.46). CONCLUSIONS: In adults, the male predominance in sensitization to HDM seems to be contrary to the prevalence of allergic airway symptoms, which has a female predominance. The male predominance is even higher in allergic sensitization to D farinae than to D pteronyssinus. Further research is necessary regarding sex-specific differences in allergy and asthma. PMID- 19558009 TI - Long-term stability of epinephrine dispensed in unsealed syringes for the first aid treatment of anaphylaxis. AB - BACKGROUND: When epinephrine autoinjectors are unavailable or unaffordable, patients at risk for anaphylaxis in the community are sometimes provided with an unsealed syringe containing a premeasured epinephrine dose for use in first-aid treatment of anaphylaxis episodes. OBJECTIVES: To study the stability of epinephrine solution in unsealed syringes under conditions of high ambient temperature, low vs high humidity, and light vs dark. METHODS: Forty unsealed syringes each containing an epinephrine dose of 0.3 mg (as a 1-mg/mL epinephrine solution) were stored at 38 degrees C for 5 months, with 10 syringes at each of 4 different standardized storage conditions: dark and light at low (15%) humidity and dark and light at high (95%) humidity. Duplicate syringes were removed monthly from each storage environment and analyzed for epinephrine content vs control syringes. RESULTS: The epinephrine dose, expressed as the percentage remaining of the mean control dose, was below compendial limits (90% to 115% of label claim) by 3 months after storage at 38 degrees C and low humidity and by 4 months after storage at 38 degrees C and high humidity. Light had no significant effect. CONCLUSION: In hot climates, if an unsealed syringe prefilled with an epinephrine dose is provided for the first-aid treatment of anaphylaxis, it should be replaced every few months on a regular basis with a new syringe containing a fresh dose of epinephrine. PMID- 19558008 TI - Suppression of histamine- and allergen-induced skin reactions: comparison of first- and second-generation antihistamines. AB - BACKGROUND: Nonsedating antihistamines (nsAHs) are recommended as first-line therapeutics for the treatment of mast cell-driven disorders, including allergic rhinitis and urticaria. However, their superiority over first-generation AHs (fgAHs) has recently been called into question, mainly because of the lack of supporting head-to-head therapeutic studies. OBJECTIVE: To compare the effects of 3 modem nsAHs with those of the fgAH hydroxyzine on histamine- and allergen induced skin reactions in a controlled, double-blind, clinical trial. METHODS: Skin prick tests with histamine and Dermatophagoides pteronyssinus extract were performed before and 4 hours after treatment with hydroxyzine, 25 mg; desloratadine, 5 mg; epinastine, 20 mg; fexofenadine, 120 mg; or placebo. Wheal and erythema development was evaluated by digital photography and planimetric analyses. RESULTS: The nsAHs prevented the development of positive reactions to histamine in only 10% to 20% of all individuals tested (n = 75). In contrast, more than 50% of all hydroxyzine-treated individuals showed negative test reactions to histamine (ie, wheals <7 mm2 in area or <3 mm in diameter). Similar differences, although less pronounced, were detected when comparing the effects of nsAHs with hydroxyzine on D pteronyssinus prick testing in a limited number of D pteronyssinus-sensitized individuals. CONCLUSIONS: These results indicate that hydroxyzine is more effective than nsAHs when given as recommended in suppressing histamine-induced or allergic skin reactions. Our results suggest that higher doses of nsAHs than those currently recommended are required for the treatment of skin responses to obtain antihistaminic and antiallergic effects that are equivalent to those of fgAHs. PMID- 19558010 TI - Home use of albuterol for asthma exacerbations. AB - BACKGROUND: To reduce symptoms and emergency department (ED) visits, the National Asthma Education and Prevention Program (NAEPP) guidelines recommend early treatment of acute asthma symptoms with albuterol and oral corticosteroids. Yet, ED visits for asthma are frequent and often occur several days after onset of increased symptoms, particularly for children from low-income, urban neighborhoods. OBJECTIVES: To describe home use of albuterol and identify factors associated with appropriate albuterol use. METHODS: A total of 114 caregivers in the intervention group of a randomized trial to reduce emergent care for low income, urban children completed a structured telephone interview with an asthma nurse to evaluate home management of their child's acute asthma symptoms. Interviews lasted approximately 20 minutes and were conducted from November 5, 2003, through September 30, 2005. Albuterol use as reported by caregivers was categorized as appropriate or inappropriate based on NAEPP recommendations. RESULTS: Albuterol use for worsening asthma symptoms was categorized as appropriate for only 68% of caregivers and was more likely if the children had an ED visit or hospitalization for asthma in the prior year. The remaining 32% of caregivers used albuterol inappropriately (overtreatment or undertreatment). Appropriate albuterol use was not associated with caregiver report of having an asthma action plan (AAP) or a recent primary care physician visit to discuss asthma maintenance care. CONCLUSIONS: Caregivers reported that they would use albuterol to treat their child's worsening asthma symptoms, but many described inappropriate use. Detailed evaluation of proper albuterol use at home may provide insight into how health care professionals can better educate and support parents in their management of acute exacerbations and more effective use of AAPs. PMID- 19558011 TI - Augmentation of bronchodilator responsiveness by leukotriene modifiers in Puerto Rican and Mexican children. AB - BACKGROUND: Ethnic-specific interactions between different asthma medications are not well described. OBJECTIVE: To determine whether the use of leukotriene modifiers is associated with the magnitude of bronchodilator responsiveness among Mexican American and Puerto Rican children with persistent asthma. METHODS: A cross-sectional study of 84 Mexican American and 192 Puerto Rican children, with persistent asthma who were aged 8 to 16 years. Within each group, bronchodilator responsiveness to albuterol, objectively assessed via spirometry, was compared between participants using leukotriene modifiers and those not using leukotriene modifiers. RESULTS: Leukotriene modifier use was associated with a clinically significant increase in percentage change in forced expiratory volume in 1 second of 11.8 (P < .001) in Puerto Rican children, but there was no significant change in percentage change in forced expiratory volume in 1 second (-3.2, P=.57) in Mexican American children. This finding persisted after controlling for the use of inhaled corticosteroids. In addition, among the Puerto Rican children, the association between leukotriene modifier use and augmented bronchodilator responsiveness was greatest in those younger than 12 years. CONCLUSIONS: Among children with persistent asthma, use of leukotriene modifiers is associated with augmented bronchodilator responsiveness to albuterol in Puerto Ricans, but not Mexican Americans. This ethnic-specific, drug-drug interaction highlights the need for the further understanding of asthma pharmacogenetics among children from different ethnic groups to improve asthma outcomes. PMID- 19558012 TI - Utility and limitations of objective measures of asthma. PMID- 19558013 TI - Proteomics methods and applications for the practicing clinician. AB - OBJECTIVE: To describe clinical proteomics from discovery techniques and their limitations, to applications in allergy, asthma, and immunology, and finally to how proteomics can be integrated into clinical practice. DATA SOURCES: Despite many inherent challenges, proteomics-based methods have become a powerful and popular means of profiling clinical samples for the purpose of biomarker discovery. Although several strategies exist, clinical proteomics for the purpose of biomarker discovery generally focuses on 1 of 3 basic workflows: (1) 2 dimensional gel electrophoresis to quantitate relative protein levels followed by mass spectrometry (MS) to identify proteins of interest, (2) non-gel-based methods that rely on liquid chromatography MS (LCMS) for both quantitation and identification of proteins, and (3) protein profiling methods that do not directly result in the identification of proteins but rather generate "fingerprints" that are compared among individuals or samples. STUDY SELECTION: Regardless of the strategy being pursued, a few general experimental steps are followed that will be expounded on in the text. These proteomics techniques have been applied to discover new biomarkers in biofluids and tissues from individuals with a variety of conditions, including allergy, asthma, atopic dermatitis, inflammatory diseases, chronic obstructive pulmonary disease, and other lung diseases. RESULTS: After biomarker discovery, LCMS-based proteomics offers several advantages over traditional antibody-based clinical assays, including greater specificity, cost- and time-effectiveness, and the potential to multiplex up to hundreds of peptides in a single assay. CONCLUSION: With many guidelines now in place and model studies on which to design future experiments, there is reason to be optimistic that candidate protein biomarkers will be discovered using proteomics and translated into clinical assays. PMID- 19558014 TI - Limitations of objective measures of asthma. PMID- 19558015 TI - Anaphylaxis due to treatment of gastric protection. PMID- 19558016 TI - Ensuring a comparable standard of care for cesarean deliveries. PMID- 19558017 TI - Automation ends preop paper chase. PMID- 19558018 TI - Computed tomographic (CT) colonography for colorectal cancer screening and diagnosis. PMID- 19558019 TI - Endo reprocessing lapses at the VA. PMID- 19558020 TI - Are you ready for Joint Commission? PMID- 19558021 TI - GPO purchasing of implants lags. PMID- 19558022 TI - Economy is taking a toll on ASCs. PMID- 19558023 TI - Automation for ASCs picking up speed. PMID- 19558024 TI - To publish or not: that is not the real pilot study question? PMID- 19558025 TI - Implementing practice changes: walk before you run. PMID- 19558026 TI - Enacting nursing obligations: Public Health Nurses' theorizing in practice. AB - A recent study illustrated Public Health Nurses' theorizing of obligation within their everyday practice with high priority families. As a practical (and practice) activity this theorizing shaped and enhanced PHN practice in complex nursing situations and served to affect desired outcomes for families. Using a hermeneutic methodology, six features were identified as central to this practical theorizing process: (a) being in the complexity of families' lives, (b) responding to relational cues, (c) putting the primary investment in people and families, (d) working toward potential, (e) working the relational "in-betweens," and (f) reflexive inquiry. The findings have implications for understanding theory and theory development in everyday nursing practice. PMID- 19558027 TI - A systematic review of questionnaires measuring health-related empowerment. AB - The objective of this systematic review was to identify questionnaires that measure health-related empowerment in adults or families and demonstrated the best evidence of reliability and validity. A search of nine data bases identified 8,269 abstracts that referred to empowerment. Full article review was completed for abstracts that met the inclusion criteria or that could not be excluded with certainty (n = 124). Fifty distinct, modified, or translated questionnaires measuring empowerment were identified in 74 articles. Each was rated in terms of reliability and validity. One questionnaire had good evidence of reliability and validity, four had moderate evidence, and 45 had limited or no evidence. Limited or no evidence for reliability and validity for many questionnaires could relate in part to lack of consensus on the theoretical definition of, and indicators for measuring empowerment. We recommend that researchers use the questionnaire rated as having good evidence and that data on reliability and validity continue to be reported for other questionnaires. PMID- 19558028 TI - Exploring possibilities: virtual reality in nursing research. AB - This article describes the use of virtual reality (VR) as a method of measurement in nursing research. Virtual reality refers to the use of computerized displays to show a lifelike environment in which the user interacts. Although many disciplines are beginning to use VR environments in research, nursing has yet to embrace this technology. Nursing, as a profession that values the interaction between the environment, individual, and health, can benefit from the use of VR in research. Establishing reliability and validity of the VR tool selected for research is important and requires special consideration. Virtual reality testing can produce side effects, such as vertigo and discomfort, which must be anticipated in the research protocol. PMID- 19558029 TI - The rhythm of health in older women with chronic illness. AB - Trends in population growth indicate that nurses increasingly will be caring for older adults, especially older women, who are at risk for less than optimal health due to chronic illnesses. The purpose of this qualitative study was to obtain a better understanding of the meaning of health from the perspective of older women. Focus group interviews with 51 women, aged 55 to 93, were held. Data were analyzed using directed qualitative content analysis. A Rogerian perspective of human health provided the broad conceptual lens for the study. Three themes explicating the meanings of health emerged from the data: realizing the potential for purpose, listening to energy flow, and purposefully participating in health related changes. The three themes represented ways in which the women dealt with the paradoxes posed by their chronic illness. Findings captured the hopeful, although sometimes conflicting, meanings of health among the women as brought forth by experiences with chronic illness and the challenges of aging. PMID- 19558030 TI - The recovery audit contractor program: an overview. PMID- 19558031 TI - Community capacity for cancer prevention. AB - We pilot-tested a street-level study of availability of physical resources to assess ethnic disparities in community capacity for cancer prevention in forty Brooklyn, NY, census tracts with high proportions of White, African American, or Jamaican immigrant populations. Interns with GIS maps made street-level inventories of food retailers, fast-food restaurants, and commercial exercise facilities. Availability was quantified as resources per capita or square mile. Median income-adjusted number of supermarkets, greengrocers and fast-food restaurants per square mile was significantly higher in Jamaican than in African American or White tracts. Bodegas per capita was greatest in African American tracts, with no significant differences among the population groups in availability of health food stores, or commercial exercise venues. PMID- 19558032 TI - Health literacy and English language comprehension among elderly inpatients at an urban safety-net hospital. AB - OBJECTIVE: To evaluate the relationship between health literacy and age in chronically-ill inpatients at a safety-net hospital. SETTING AND PARTICIPANTS: We recruited 399 English- and Spanish-speaking inpatients being evaluated or treated for Congestive Heart Failure or Coronary Artery Disease at a large, urban safety net teaching hospital in Southern California. DESIGN: Participants were interviewed to ascertain education, English comprehension, and in-home language use. Health literacy was assessed using The Test of Functional Health Literacy in Adults (TOFHLA). We compared by age (aged 65 or more, 51 to 64 years of age, and less than age 50) levels of health literacy, educational attainment, English comprehension, and language use. RESULTS: Prevalence of inadequate health literacy significantly increased with increasing age (87.2% in > or = 65, 48.9% for 51-64, and 26.3% in < or = 50, p<0.001). The correlation between older age and lower health literacy persisted when controlling for educational achievement, race, ethnicity, gender, and immigration status. Additionally, older patients were more likely to have never learned to read (34.9% in > or = 65, 6.5% for 51 64, and 1.5% in < or = 50, p<0.001), no formal education (27.9% in > or = 65, 9.0% for 51-64, and 0.8% in < or = 50, p<0.001), have limited English comprehension (74.2% in > or = 65, 43.5% for 51-64, and 35.8% in < or = 50, p<0.001), and speak a non-English language at home (82.3% in > or = 65, 70.2% for 51-64, and 62.2% in < or = 50, p=0.015). CONCLUSIONS: To prepare to meet the chronic disease needs of a growing older patient population, and ameliorate the negative health effects of associated low literacy, safety-net hospital leaders and providers need to prioritize the development and implementation of low literacy educational materials, programs, and services. PMID- 19558033 TI - Knowledge, behaviour, perceptions and attitudes of University of Ghana students towards HIV/AIDS: what does behavioural surveillance survey tell us? AB - This study assesses knowledge, attitudes and behaviour in respect of risk of HIV infection of students through behavioral surveillance survey. The study used the systematic sampling approach to select 375 students. Interviews and focus group discussions were conducted to solicit information from respondents. The study found out that the students engaged in pre-marital sex, although this was more common among the male than female students. Students did not use condoms consistently and were not likely to use condoms when the relationship was considered as stable because of trust. Students revealed that they were all at risk of infection; however, a significant number of them did not know their HIV status and were not ready to take the test for fear of stigmatization. There low level knowledge among female students of the different types of STI. The results suggest that there is need for an appropriate education program for students. Peer education clubs need to be established to provide education on condom management. In addition, edutainment-education through entertainment program on HIV/AIDS -- needs to be an integral part of the university's curriculum. More recreational centers need to be established to allow students to channel their energies towards sports. PMID- 19558034 TI - Drug treatment program compliance and resistance activities during implementation of California's Proposition 36. AB - The Substance Abuse and Crime Prevention Act (SACPA), also known as Proposition 36, was implemented statewide in 2001 in California. This legislation remands non violent drug offenders to drug treatment rather than prison or jail. Structured telephone interviews were conducted with a purposive sample of 72 drug treatment programs from six Southern California counties concerning compliance and resistance activities during implementation. A linear regression model was developed that used a dependent variable capturing overall experience with Proposition 36 and compliance and resistance activities as independent variables. The final model included three compliance variables (sharing problems and solutions with other treatment providers, hiring new staff, acquiring additional space through rental or purchase) that were most predictive of the programs' overall experience with Proposition 36. The implications of these findings in the context of organizational compliance and resistance activities are discussed. PMID- 19558035 TI - The effects of involvement and ad type on attitudes toward direct-to-consumer advertising of prescription drugs. AB - This article examines consumers' attitudes toward Direct-to-Consumer (DTC) advertising of prescription drugs that are influenced by the use different types of DTC ads and product involvement. Our findings suggest that product involvement and the type of DTC ad are significant predictors of consumers' attitudinal responses toward DTC advertising. High involvement consumers have more favorable attitudes toward the drug's price, DTC ad and brand name, and a higher intention to ask a doctor about the advertised drug than low involvement consumers. In contrast to Informational and Reminder DTC ads, Persuasive ads have more favorable effects on consumers' reactions to DTC prescription drug advertising. PMID- 19558036 TI - [Endocrine disruptors -- Monograph. 3. Arsenic]. PMID- 19558037 TI - [Psychoactive drugs and fitness to drive in the occupational field]. AB - According to/After the conferences taken place on 30th October and on 18th December 2008, the company doctor functions have been enlarged: at present they include the duty of effectuating toxicological tests in order to find psychotropic and drug substances in some specific working classes, mainly in workers employed in public transports. Company doctors have the duty to report any possible positive matching in these workers'tests. From there, regulations in force are analyzed focusing to critical situations described in the complex and well articulated frame of reference. PMID- 19558038 TI - [Extra-auditory effects of noise]. AB - In the last thirty years, several studies have demonstrated a correlation between exposure to noise and the development of alterations and/or pathologies in organs and apparatus, apart from the auditory one. Exposure to noise can induce biochemical, physiological, or psychosocial modifications that can remain either inside the range of biological normality or alter and compromise the psychophysical well-being of the individual depending on the extent of the functional alterations of organic systems and psychosocial functions; both the reversibility and duration of the alterations and the adaptation could be related to the recovery capabilitiy of the human organism and to the environmental condition. Among the factors that can influence the effects of noise we can find: intrinsic characteristics of the physical insult (pressure, sound intensity, emission frequency), extrinsic characteristics (duration of the exposure, way of emission, presence of impulsive components, masking effects), spectral characteristics (infrasounds, ultrasounds, time of recovery) genetic factors (susceptibility) and acquired factors of the human organism (acoustic isolation of the living houses and the social necessity for the industrial activities that generate noise). Other factors that influence the damage are the surprise effect, the semantic content and the identification of the noise source. It can be hypothesized that the extra-auditory effects of noise show themselves through a series of nervous circuits that use the autonomic nervous system and interfere on neuro-immune-endocrine parameters. The organs which are more studied are: cardiovascular, gastro-enteric and endocrine systems, nervous system and psychological effects, respiratory system, fetus and effects on reproductive system, immune system and genetic material. PMID- 19558039 TI - [On confusions]. PMID- 19558040 TI - [New molecular indicators for the prevention of tumor and degenerative diseases: anomalous DNA methylation]. AB - DNA methylation is a physiological mechanism regulating both gene expression and genomic stability. Abnormal DNA methylation is observed in neoplastic as well as degenerative disease, and may affect both the whole genome and specific genes. Aberrant DNA methylation has been recently associated to the exposure to carcinogenic chemical compounds. The characterization of DNA methylation abnormalities by modern molecular biology methods may allow the development of new molecular biomarkers of early biological effects. PMID- 19558042 TI - [Occupational therapy]. PMID- 19558041 TI - [Mechanisms of action for metallic elements and their species classified as R40 by EU]. AB - In this part of our survey we dealt with other metallic elements and species (antimony, lead and vanadium) classified by EU as R40 and not with the risk phrases R45 and R49. They are also differently considered by different Agency and Scientific Societies in respect to their carcinogenicity. Also for Pb, Sb, V and related species the carcinogenic effect is related with oxidation state, charge, the solubility, type of binding, stereochemistry. Some common mechanisms of carcinogenesis are the induction of oxidative stress, to inhibition of DNA repair, from activation of mitogenic signalling, to epigenetic modification of gene expression and each species lead to specific molecular interactions and were subject to different bioavailability. We focused in particular the mechanisms of action for lead, element moved in last IARC examination from group 2B to group 2A, with important potential consequences for EU classification In general for metallic elements even differently classified in respect to their carcinogenicity, knowledge of action mechanisms would give additional tools to reach more adequate risk assessment procedures and the preventive-health surveillance measures. PMID- 19558043 TI - [Environmental control and alternative augmentative communication in severe motor disability]. PMID- 19558044 TI - [Physical component of rehabilitation in complex cardiopathy]. PMID- 19558045 TI - [Physical reconditioning of critical respiratory patients]. PMID- 19558046 TI - [Posture and movement laboratory: from posturography evaluation to rehabilitation treatment]. PMID- 19558048 TI - [Instrumentation for assessing care needs and nursing in rehabilitation]. PMID- 19558049 TI - Nursing and the great recession. PMID- 19558047 TI - [Robotics in rehabilitation]. PMID- 19558050 TI - Towards clarification of the role of research nurses in New Zealand: a literature review. AB - The demand for research nurses has increased markedly in recent years due to a rapidly expanding clinical research environment. Research nursing is becoming increasingly specialised and nurses now hold central positions in the coordination of clinical trials and management of trial related patient care. However, as a role in transition, research nursing is currently under-recognised and has yet to establish a clear identity and position for itself within nursing and within research. Therefore, the aim of this literature review is to describe the current role of research nurses in clinical trials and explore the professional issues surrounding the role. To provide greater clarification of the role in New Zealand, these findings are examined against the Nursing Council of New Zealand competency requirements for registered nurses. In examining the professional issues surrounding the role, current barriers to adequate recognition of the role are identified and potential strategies to clarify the role and the position of research nurses are put forward. PMID- 19558051 TI - The effects of staff education on the practice of 'specialling' by care assistants in an acute care setting. AB - This paper reports the results of a project aimed at decreasing the use of, and costs associated with, the practice of using care assistants to provide one-to one observation or 'specialling' in five acute adult medical and surgical wards at a North Island hospital. Education sessions were provided to staff to prompt better management of patients receiving this level of observation. Using a time series design, a retrospective study was completed for the six months pre- and six months post-intervention (staff education). Data were collected and analysed on all patients in the five acute wards who had required 'specialling' during the specified twelve-month period. Results showed that following the intervention there had been a reduction in the incidence and duration of 'specialling,' and a halving of costs associated with this practice. The findings have implications for patient management and cost effectiveness. Incident statistics were not retrospectively reviewed in this study. PMID- 19558052 TI - Innovative nursing leadership in youth health. AB - Along with introducing the New Zealand Primary Health Care Strategy the Ministry of Health funded 11 primary health care nursing innovation projects for three years. One of these was sited in the Hutt Valley District Health Board region at what was originally known as the Hutt Valley Youth Service and later named the Vibe service. The funding supported employment of a clinical nurse leader who later, during the course of the project, qualified as a Nurse Practitioner (NP). The dynamic leadership provided by the nurse extended beyond service boundaries benefiting policy decision making throughout the District Health Board region and contributing to youth health and service development nationally. In return for fostering such leadership Vibe gained access to new dimensions of youth health care. It became apparent that high calibre clinical leadership, a major component of the NP scope of practice in primary health care, improves outcomes for the clientele, increases satisfaction for the team members and provides a model for potential NPs wherever they are situated. PMID- 19558053 TI - The impact of a glaucoma nurse specialist role on glaucoma waiting lists. AB - Over the last two decades there has been an increasing demand on health care services. This has led to increased waiting lists and waiting times to access public hospital services. In ophthalmic practice an aging population and technological advances have been major contributors to this situation. The challenge for health care providers is how to manage waiting lists within the resources provided and targets set by Elective Services, a department of the Ministry of Health (MoH). The Ministerial Taskforce on Nursing (1998) highlighted that nurses are an underutilised resource. Subsequent to the Ministerial Taskforce, the availability of interim funding from Elective Services and a skilled speciality nursing workforce the Ophthalmology Department at a large metropolitan hospital was able to implement an advanced nursing role with respect to glaucoma management. Early diagnosis is important in reducing the risk of permanent visual impairment from primary open angle glaucoma; therefore the length of time to diagnosis can have long term implications for the patient. Patients for the 'nurse-led' glaucoma clinic were recruited from specific categories of glaucoma patients on the waiting list. An audit of the impact on the waiting list was undertaken periodically over a two year period. The audit following the introduction of the clinical nurse specialist clinic shows a marked reduction in waiting list numbers and length of time waiting for first specialist assessment for glaucoma. The implementation of this role proved to be successful and has subsequently become a permanent nurse specialist position in the ophthalmology department. PMID- 19558054 TI - Re: Editorial vol 24, no 3, 2008. PMID- 19558055 TI - Ocular evaluation in patients with chronic renal failure--a hospital based study. AB - Chronic renal failure affects every organ system including eye. The aim of this study is to conduct thorough ocular examination in the patients of chronic renal failure and to analyze the findings. 119 cases were collected from Nephrology unit of Tribhuvan University Teaching Hospital between 1st June 2002 to 15th December 2003. This was a cross sectional, descriptive type of study. Sampling technique was consecutive and stratified. Severity of renal disease was classified as mild, moderate, severe and end stage renal disease. Twenty-three percent of total 238 eyes had vision < 6/18. The causes for visual impairment were maculopathy 23 eyes, cataract 14 eyes followed by proliferative diabetic retinopathy, 9 eyes. Twelve percent of total eyes had vision < 6/60. Lid edema was present in 63.0%, conjunctival pallor in 75.6% and corneal calcification in 1.6%. Retinopathy was the most important finding. Hypertensive retinopathy was present in 56 out of total 119 cases (47.1%). It was more prevalent and tended to be more severe as renal disease progressed. This was statistically significant. Diabetic retinopathy was present in 38 out of 43 diabetic cases (88.3%). Although statistically not proven, more severe grades of diabetic retinopathy were detected with increasing severity of the renal disease. There was one case of bilateral serous detachment of the retina relating to chronic renal failure. In this study, 47 out of 56 cases of hypertensive retinopathy and 19 out of 38 cases of diabetic retinopathy were detected for the first time, thus showing the importance of ocular evaluation of the patients of renal insufficiency. PMID- 19558056 TI - Age and sex related variations in corpus callosal morphology. AB - There is no unanimity in literature on sexual dimorphism or on age related morphophometric changes in the Corpus Callosum (CC). For comprehensive data 44 preserved human brains (22 male and 22 female) and 30 MRI scans from North-west Indian population were studied. Morphometric measurements of the CC and its subregions were taken at the midsagittal level in both the groups and subjected to statistical analysis. The only CC parameters which showed sexual dimorphism were (a) larger CC length in males in the MRI group, (b) the distance between the splenium and superior colliculus was more in males in preserved brains and (c) The distance between the genu and the fornix was more in older males than older females. Age related changes included (a) In males the distance between the genu and the fornix (in both the groups) was more in the older age group while the maximum width of the anterior half of the CC body was more in the younger age group (preserved brains) and (b) In females the height of the CC and the minimum width of the CC were more in the older females (preserved brains). No sexual dimorphism was observed in most of the CC parameters studied, including the splenium. Age related thinning of the anterior half of the CC in males could possibly be related to atrophic brain changes more common in males than females. PMID- 19558057 TI - Role of systemic anticoagulation in patients undergoing vascular access surgery. AB - The role of per-operative systemic heparin to improve primary patency rate of vascular access surgery is controversial. The aim of this study was to assess the risk and benefit of systemic heparin during creation of vascular access for hemodialysis in patients with chronic renal failure. Patients undergoing creation of side to end radio-cephalic arteriovenous fistula over distal forearm for hemodialysis were prospectively randomized into two groups. First group received 5000 IU of intravenous heparin during surgery whereas second group did not receive any anticoagulation. Post-operative complications and outcome of surgery were compared between the two groups. Among 50 patients, 25 received heparin and 25 did not. Although there was no significant difference in operative times between these two groups (p = 0.24), early post-operative bleeding complication was more common in patients receiving heparin (p < 0.01). The primary 6-week patency was 96.0% for patients receiving heparin and 92.0% for those not (p = 0.46). Thus per-operative systemic anticoagulation during vascular access surgery is associated with increased incidence of bleeding complication and offers no benefit in terms of primary patency PMID- 19558058 TI - Role of repeat CT scan head in initially inoperable cases of traumatic head injury. AB - The CT scan has become popular in cases of head injury. In this study 71 cases (M:48; 68.0% and F:23; 32.0%) with traumatic head injury admitted and initially managed conservatively at National Neurosurgical Referral Centre, Bir hospital starting from May 2005 to April 2006. The most common cause of injury was road accidents (43 cases) with motorcycle and bus injuries as the leading causes. Nausea and vomiting were the most common symptom in 41 (57.0%) cases, followed by headache in 31 (43.0%). Contusions were the most common radiological findings in 84.0% followed by extradural haematoma in 8.0% and pneumocephalus in 7.0%. Out of these the most common location for contusion was frontal (16 cases), followed by parietal (12 cases) and then bilateral contusions. The mean volume was 12 ml, 9 ml and 9 ml for frontal, parietal and temporal contusions, respectively. Repeat scan showed increase in volume of contusion in 31 cases (44.0%), no change in 28 cases (40.0%) and decrease in 12 cases (16.0%). The increase was mainly due to edema in 20 cases (64.0%) and due to actual increase in contusion in only 11 cases (35.0%). Only 5 (7.0%) cases needed operative intervention. It is concluded that routine use of repeat scan in mild to moderate head injury has no role unless there is clinical deterioration. PMID- 19558059 TI - Phylogenetic variation in flexor digitorum brevis: a Nepalese cadaveric study. AB - Understanding muscle architecture of the foot may assist in the design of surgical procedures such as tendon transfer, biomechanical modeling of the foot, prosthesis design, and analysis of foot function. Flexor digitorum brevis (FDB) is the most superficial intrinsic muscle of the sole of the foot. When compared to the little finger, the little toe has less function and opposition is absent in humans. Hence, the muscles acting on the little toe are undergoing evolutionary changes. The objective of this study was to review the phylogenetic variation occurring in FDB in the Nepalese cadavers. Sixty soles in 20 male and 10 female adult embalmed cadavers were dissected and attachments of FDB observed. It was noted that the tendon for the fifth toe was missing in all sixty soles. The action of FDB can be compensated by other long flexors of the foot. This could have resulted due to gradual reduction in the usage of the little toe as the bipedal posture evolved. Variations of FDB are clinically important because FDB musculocutanous flap is used in the reconstruction of the heel pad. PMID- 19558060 TI - Epidemiological and bacteriological profile of burn patients at Nepal Medical College Teaching Hospital. AB - This study was conducted to know the epidemiological and bacteriological profile of burn patients at Nepal Medical College Teaching Hospital. The charts of 50 burn patients admitted in department of surgery were reviewed retrospectively. All the epidemiological characteristics, mode of injury, time taken to reach hospital and involved body surface areas were noted. The charts were also reviewed for bacterial isolates from burn wounds and its sensitivity pattern for various antibiotics. Data was analyzed using the statistical package for social sciences (SPSS) for Windows. The mean age of patients was 31.8 years with male:female ratio of 1.3:1. Half of the patients were from Kathmandu. Fifty two percent of patients directly came to our hospital while rests were referred from other hospitals. The average time taken to reach hospital was 11.3 hours. Those patients who were referred from outside the valley took longer time (p = 0.002). Flame burn was the leading cause for injury (66.0%) followed by scald burn (16.0%), electric burn (14.0%) and acid burn (4.0%). Staphylococcus aureus (28.0%) was the commonest organism isolated from wound swab culture. Others were Klebsiella (16.0%), Pseudomonas (13.0%), Proteus (13.0%) and E.coli (13.0%). No growth was noted in 17.0% of patients. During the treatment, 14.0% of patients died and 4.0% left against medical advice. Remaining patients were discharged after complete recovery. Body surface area involvement was found to be a significant predictor of mortality (p < 0.001) and the length of hospital stay was significantly low for them (p = 0.05). PMID- 19558061 TI - Prevalence of Salmonella typhi and paratyphi infection in children: a hospital based study. AB - The purpose of this study was to find out the prevalence and antibiotic sensitivity pattern of Salmonella typhi and paratyphi isolated from children presenting with fever at Kanti Children's Hospital in Kathmandu, Nepal. A total of 9,856 blood samples collected for culture during one year period (April 2007 to March 2008) were included in the study. Out of total, 235 (2.0%) were positive for S. typhi and paratyphi A. Of the total positive, 195 (83.0%) were S. typhi and 40 (17.0%) were S. paratyphi A. The growth positive rate in two genders (M: 53.2% and F: 46.8%) was not significant (P > 0.05). Over two-third of cases were clustered in the age-group of 1-10 years. The occurrence of infections was common in summer months (rainy season). S. typhi was found to be most sensitive to cefotaxime (100.0%) followed by ceftriaxone (98.9%), ofloxacin (93.5%), cotrimoxazole (93.5%) and chloramphenicol (93.2%) and was least sensitive to amoxyccilin (66.7%) followed by ciprofloxacin (86.6%). S. paratyphi also was found to be most sensitive to cefotaxime (100.0%), followed by ceftriaxone (97.4%), cotrimoxazole (97.1%) and chloramphenicol (92.5%) and was least sensitive to amoxycillin (15.0%) followed by ciprofloxacin (51.3%) and ofloxacin (70.3%). PMID- 19558062 TI - Microalbuminuria among type-2 diabetes mellitus patients in Pokhara, Nepal. AB - Microalbuminuria is considered to be an early stage of diabetic nephropathy as well as a marker of cardiovascular disease. The aim of this study was to see the prevalence of microalbuminuria in type 2 diabetic patients and assess its association with cardiovascular risk factors among them. A total of 143 type 2 diabetic patients with the mean age of 56.06 +/- 1.08 years were analysed. The prevalence of microalbuminuria and overt proteinuria was 45.5% and 11.2%, respectively. Prevalence of microalbuminuria in female was marginally higher than in male (p > 0.05). Subjects with microalbuminuria had significantly higher blood pressure (p < 0.001) and duration of diabetes (p < 0.05) compared with normoalbuminuric subjects. High density lipoprotein was found to be significantly lower (p < 0.05) in subjects with microalbuminuria whereas fasting blood sugar, triglyceride, total cholesterol and very low density lipoprotein were marginally higher in microalbuminuric than in normoalbuminuric subjects (p > 0.05). High prevalence of microalbuminuria in diabetic patients and its positive association with blood pressure and altered lipid profile suggests that screening for microalbuminuria is essential for intervention and prevent further complications like end stage renal disease and cardiovascular diseases. PMID- 19558063 TI - Evaluation of postural hypotensive subjects by handgrip test. AB - Postural syncope may be noticed in otherwise normal persons with defective postural reflexes. Vasoconstrictor reserve--the extent to which total peripheral resistance can be increased from baseline condition is an important determinant of orthostatic tolerance. Women showed low autonomic support of blood pressure. The case of neurally mediated syncope is known to be greater in females in comparison to males. The present study was designed to compare vasoconstrictor reserve in male and female participants having no complaint of postural hypotension and the vasoconstrictor reserve in female volunteers having complaints of occasional postural hypotension. Vasoconstrictor reserve was determined by asking the subjects to sustain isometric handgrip at one third of maximum handgrip contraction for one minute using a handgrip dynamometer. Vasoconstrictor reserve was noted as difference between the maximum diastolic pressures at the end of one minute of sustained isometric handgrip minus baseline diastolic pressure. Results of the present study indicated less vasoconstrictor reserve in females compared to their male counterparts. When compared, the vasoconstrictor reserve of female participants experiencing occasional postural hypotension with that of the normal female volunteers, it was noticed that the same in them was much lower than that in normal female volunteers. This less vasoconstrictor reserve might be one of the causes of postural hypotension among the women participants experiencing it in our study population. PMID- 19558065 TI - Single burr hole drainage for chronic subdural haematoma. AB - Chronic subdural haematoma is a common neurosurgical condition and surprisingly surgical treatment ranges from twist drill craniostomy to more radical membranectomy. However, the outcome is generally favourable with appropriate therapy. This retrospective study is aimed at analyzing the result of single burr hole drainage of such haematoma under local anaesthesia over a period of fifteen years. There were a total of 365 patients ranging from 6 months to 89 years with the mean age of 60 years. Recurrence of haematoma was noted in 17 (4.6%) patients and majority of them (65.0%) were managed by aspiration through the previous burr hole. Favorable outcome was noted in 98.6% patients. Those with adverse outcome were in coma preoperatively. This study suggests that single burr hole drainage under local anaesthesia is sufficient in the majority of patients and outcome was favorable even in the elderly provided they presented before lapsing into coma. PMID- 19558064 TI - Assessing placement of nasoduodenal tube and its usefulness in maintaining nutrition in critically ill patients. AB - Nutritional supplements to the critically ill patients are one of the major issues to be discussed. Enteric feeding is advantageous over parentral feeding because it maintains gut integrity and prevents bacterial translocation. Small intestinal feeding shows significant beneficial results. Nasoduodenal tube placement and its confirmation by serial pH monitoring and by radiological examination was done; and time taken to reach duodenum was assessed. This study was done in 40 critically ill patients, who were divided into two groups (Group 1 and Group2). Early enteric feeding via a nasoduodenal tube was found to be preferable with parentral therapy when there are no contraindications. Advantages of nasoduodenal tube feeding in critical ill patients were that feeding can be continued even in the absence of bowel sounds and passage of flatus. Insignificant complications were noted. PMID- 19558066 TI - Patients with tonsil problems at NMCTH--a report. AB - The study was carried out in the Department of Otorhinolaryngology NMCTH (n = 20, age 3-54yrs) who presented tonsil problems and underwent tonsillectomy over period of 1 year (2006-2007). Patients who underwent tonsillectomy were selected for study throughout preoperative and two months of postoperative period. Out of 20 patients (8 male andl2 female) 3 patients presented with sleep apnoea syndrome, 8 patients required hospitalization due to odynophagia and high fever and 9 of them suffered from recurrent tonsillitis. Post operative period was uneventful. The commonest presentation was recurrent sore throat and commonest organism isolated on throat swab culture (superficial) was S. aureus (26.0%). Sixty-nine percentage of culture was sterile, most probably because of prior antibiotics use. Only one out of twenty cultures showed a haemolytic streptococcus. According to previous studies carried out, organism obtained from superficial swab may be different from that of from core and there were some differences between children and adults. a haemolytic streptococci was the commonest organism (38.0%) isolated from surface. PMID- 19558067 TI - HELLP syndrome--a pregnancy disorder with poor prognosis. AB - HELLP syndrome is a pregnancy-specific disorder defined by hemolysis, elevated liver enzymes and low platelet count that is found in parturients, more frequent in older multiparas. It is frequently associated with severe preeclampsia or eclampsia, but can also be diagnosed in the absence of these disorders. The etiology of HELLP syndrome is unknown, and the pathogenesis of this disorder (including the hepatological manifestations) is not fully understood. The most widely accepted hypotheses are: a change in the immune feto-maternal balance, platelet aggregation, endothelial dysfunction, arterial hypertension and an inborn error of the fatty acid oxidative metabolism. Hepatic involvement occurs by intravascular fibrin deposition and hypovolemia. Serum LDH and platelet count are the two most important clinical tools for disease assessment. LDH reflects both the extent of hemolysis and hepatic dysfunction. Maternofetal complications cause a 7.0-70.0% perinatal mortality rate and a 1.0-24.0% maternal mortality rate. The recognition of HELLP syndrome and an aggressive multidisciplinary approach and prompt transfer of these women to obstetric centers with expertise in this field are required for the improvement of materno-fetal prognosis. PMID- 19558068 TI - Relationship between blood group and epistaxis among Nepalese. AB - The relationship between blood group and epistaxis among the patients coming to Tribhuvan University Teaching Hospital, Kathmandu with idiopathic epistaxis was studied. A total of 235 patients coming to the Emergency Department or Department of Otorhinolaryngology with active nose bleeding in were included. ABO blood group was performed. Nearly half (45.5%) of the patients belonged to O-group followed by A-group others. O-group people were also had longer bleeding time compared to other blood groups. PMID- 19558069 TI - Bilateral slipped capital femoral epiphysis: a case report and review of literature. AB - Slipped capital femoral epiphysis (SCFE) is a common adolescent problem in the western populations but it is extremely rare in the Asian regions. We report a case management of a teen age boy (16 years old at the time of operation) with bilateral SCFE. After fall injury, the boy sustained SCFE on the left side and was treated. Again after six months with trivial injury he got SCFE on the right side. In both cases the boy was managed by closed reduction and percutaneous cannulated cancelleous screws fixation under c-arm fluoroscopy control. The cannulated screws were removed after one year of insertion confirming of a good fusion in plain x-ray films. During the regular follow up of up to one year the patient had good hip functions and other daily activities. The patient lost to follow up for about three years and suddenly presented to us with restricted hip joints movements and other activities. The x-ray showed tilted pelvis, reduced, incongruent hip joint spaces with malunited femoral physis, widened neck and tilted femoral head along with visible physeal scan on both sides. The patient and patient party are still satisfied with these results and reluctant to go for further surgical intervention. PMID- 19558070 TI - Blind nasal intubation in a child with ankylosis of temporomandibular joint. AB - Blind nasal intubation has been one of the favoured methods of intubating the trachea in patients with restricted mouth opening and thus a difficult laryngoscopy and oro-tracheal intubation. Here, we describe airway management of patient who presented with traumatic ankylosis of the right temporomandibular joint and reduced mouth opening that was planned for elective release of the ankylosis and temporalis fascia flap. Successful blind nasotracheal intubation was done and after the surgery was completed, awake extubation was done and mouth opening was significantly increased. The recovery and the post-operative periods were uneventful. PMID- 19558071 TI - Pancreatic metastases of renal cell carcinoma--case report. AB - Metastasis to the pancreas from renal cell carcinoma is distinctly uncommon. A solitary renal cell carcinoma (RCC) metastasis to the head of pancreas is rarely encountered. We present a 55 years old male who was admitted in our hospital with the complaint of abdominal pain on and off but more continuous since 15 days. He did not have any urinary symptoms. The laboratory tests were within the normal limits. Ultrasound of abdomen revealed an encapsulate mass in the kidney plus a mass in the head of pancreas, but no other distant metastases. On histopathological examination, sections of the renal mass showed features of a highly vascularized, clear cell neoplasm, with solid, nested, and alveolar architecture, consistent with RCC, clear cell type. Fine needle aspiration from the mass in the head of the pancreas confirmed the diagnosis of metastatic renal cell carcinoma, clear cell type. In the absence of widespread disease, pancreatic resection can provide long term survival in metastatic RCC, although few cases have been reported with lengthy follow up. The prognosis is better than for pancreatic adenocarcinoma. PMID- 19558072 TI - Complex regional pain syndrome (CRPS) type-1 following snake bite: a case report. AB - The pathophysiological mechanism and clinical course of complex regional pain syndrome (CRPS) type-I still remain ill defined. Both the treatment and the prediction of the outcome of the treatment are difficult. Abnormal neurohumoral and inflammatory mechanisms have been implicated in its causation usually following trivial noxious event in an extremity. However, to the best of our knowledge CRPS type-1 following snakebite has not been reported yet in the literature. We here report a case of an aggressive CRPS type-1 following a mountain pit viper bite, locally known as Gurube (Ovophis monticola monticola) in a 55-year-old lady. The clinical condition responded well to the therapy with serial sympathetic blockade of the limb with local anaesthetics, non-steroidal antiinflammatory analgesic, antiepileptic, antidepressant and physiotherapy. Our experience in managing this patient and associated pathophysiology in development of CRPS type-1 are discussed. PMID- 19558073 TI - Nursing on the right side: why nurses must step to the right (of our left brains). PMID- 19558075 TI - Patterns of knowing: proposing a theory for nursing leadership. AB - In a time of chaotic and unpredictable health care, it is vital for nursing to employ a nursing leadership theory that is specifically applicable to nurses and will holistically, and comprehensively address and support both the science and art of this honored profession. The authors propose that Nursing Leadership Knowing can address and impact the myriad issues confronting managers and administrators within the turbulent health care industry, with the ultimate goals of quality comprehensive patient care and improved employee satisfaction. They believe that Nursing Leadership Knowing, grounded in the realties of nursing experience, is a logical theoretical extension that can be translated into nursing leadership practice particular and specific focus on empirics and evidence-based practice will not attend to the robust and multidimensional underpinnings of the lived experience that is so vital to nursing as a caring profession. The ideal of nursing leadership theory is not a single-focused shadow of its history, but a rich, inclusive, multi-faceted network of knowing. As such, Nursing Leadership Knowing provides a forum for leaders to enhance their practice, as well as their relationship with their employees, which ultimately translates into optimal care for the patients we serve. PMID- 19558074 TI - Nurse residency programs: an essential requirement for nursing. AB - The number of new graduates who will be needed to fill positions in our acute care hospitals is astounding. The hiring and precepting of this many inexperienced nurses will severely tax hospital resources. A sound plan must be developed to maintain quality of care and patient safety with the influx of so many new nurse graduates. New graduates also must have a positive learning experience in order to keep them in the nursing workforce. A residency program is essential for new graduates. The Centers for Medicare and Medicaid Services must step to the plate and support accredited nurse residency programs with pass through dollars. PMID- 19558076 TI - Evaluation of a nurse practitioner-led care management model in reducing inpatient drug utilization and cost. AB - The aim of this research was to evaluate the economic impact of a collaborative nurse practitioner (NP) care management model on the use of pharmaceutical resources, with a focus on antibiotics, among general medicine inpatients. Although studies have shown the effectiveness of care management by NPs, especially as reflected on length of stay and hospital cost, little is known about their impact on drug cost. The researchers utilized pharmaceutical claims data of 1,200 subjects who participated in the Multidisciplinary, Physician, and Nurse Practitioner Study from 2000 to 2004 to assess the effect of the NP-led care management model on drug utilization outcomes. Study findings revealed that the intervention group was associated with significant reduction in drug cost and drug utilization; even though the intervention group was more likely to be given broad-spectrum or other antibiotics, its overall drug and antibiotic costs were lower than the control group. Drug management strategies such as de-escalation and intravenous-to-oral conversion facilitated by NPs may potentially produce both clinically and economically advantageous outcomes among general medicine inpatients. PMID- 19558077 TI - What nurses want: the nurse incentives project. AB - Today's nurse executives are struggling with leadership challenges of managing the multigenerational workforce, financial imperatives to deliver better care for lower costs, and competition to provide the optimal work environment to retain nurses. The purpose of the Nurse Incentives Project was to determine satisfaction with current employment incentives and potential managerial actions which might decrease or delay turnover by registered nurses. This study spawned recommendations regarding the role of incentives in designing an environment where benefits and perks will be seen as incentives to stay and thrive in the current nursing workplace. The results show that nurses know what they want. Attention to generational priorities and flexible benefits programs may help to create the cohesive work environment that nurses seek. Investment into creating delivery arenas where satisfied nurses are caring for satisfied patients is a worthwhile goal. PMID- 19558078 TI - Predicting factors related to nurses' intention to leave, job satisfaction, and perception of quality of care in acute care hospitals. AB - High nursing turnover is a problem facing many countries, including Taiwan, and can be very challenging for nursing leaders and hospital administrators. More than one-third of the nurses participating in this study considered leaving their current jobs. Understanding what factors influenced nurses to leave their jobs should be of interest to nursing leaders and hospital administrators in today's changing health care environment. Factors related to intention to leave, including wage dissatisfaction and work shift, should be addressed by hospital administrators. The findings can help hospital administrators and nursing leaders to develop strategies that minimize nurses' intention to leave behaviors, stabilize the nursing workforce, maintain hospital-employee relationships, and prevent nurses from experiencing burnout. PMID- 19558080 TI - Supplemental recruiting services: a viable solution in challenging times. AB - The need to hire new or replacement staff, especially for hard-to-fill positions, does not disappear, regardless of the economy or budget constraints. Supplemental recruiting services can augment your hospital's existing HR and recruiting resources. To offset reductions in HR staff, bringing supplemental recruiting resources on board during an aggressive hiring initiative to fill the recruiting pipeline and augment existing HR resources is both cost effective and prudent. Using an external, expanding recruiting team can be beneficial for the bottom line and viable for a changing set of circumstances. Up markets and down, these services provide value, flexibility, and viable assistance. PMID- 19558079 TI - Churn: patient turnover and case mix. AB - Patient throughput and casemix changes on nursing wards are little understood aspects of nursing's responsibility for nursing wards/units as hospital operations. In this study, the movement of patients on and off wards in 27 Australian public hospitals (286 wards) were analyzed over a 5-year period. Casemix change at the nursing unit level was also examined. In the data here, medical/surgical patients moved on average more than twice in an average hospital stay of only about 4 days. The absence of ward-level metrics compromises the ability of nursing unit/ward managers to meet their own efficiency and quality standards. Measurements of churn would give nurses another way to talk about the work of nursing to senior management and would give nurse executives a way to describe hospital operations and throughput and the impact on staff, patients, and resource allocation. PMID- 19558081 TI - Massachusetts health care reform: lessons for the nation? An interview with Thomas Smith, MS, RN, NEA-BC. Interview by Peter I Buerhaus. PMID- 19558082 TI - Improvement in prescription renewal handling by application of the Lean process. AB - The health care industry, challenged to deliver improved quality of care at lower cost, may benefit from application of Lean methodologies used by other industries to meet similar challenges. Management of phone requests for renewals of prescriptions at this ambulatory care setting was wrought with delays, extra steps, and ineffective work practices. Lean methodologies assisted in identifying a prescription renewal process with more value and less waste, mbulatory care leaders and staff may successfully apply Lean methods to achieve cost-effective care with improved quality of care for many of the care and business processes. PMID- 19558083 TI - The virtue script. PMID- 19558084 TI - ALERT: Revatio is another brand name for sildenafil. AB - In this column, the authors highlight a medication incident that occurred with Revatio (sildenafil), along with the learnings and recommendations from a previously published ISMP Canada Safety Bulletin. It is well-known to health care practitioners that use of nitroglycerin therapy is contraindicated in patients taking sildenafil (commonly known as Viagra). Many health care practitioners may be unaware that sildenafil is also marketed under the brand name Revatio for treatment of primary pulmonary hypertension or pulmonary hypertension secondary to connective tissue disease. The following incident signals the need to heighten the awareness that Revatio is a brand name for sildenafil. PMID- 19558086 TI - [Distribution character of polycyclic aromatic hydrocarbons in soils from the karst cavity, Guilin City, South China]. AB - We investigated the distribution characteristic of polycyclic aromatic hydrocarbons (PAHs) of soils from Dayan cave, a karst cavity in Guilin City, South China. The total PAHs concentrations from the inner cave ranged from 7.22 to 117.29 ng x g(-1) were much lower than those of outside cave (range of 51.35 235.73 ng x g(-1)). PAHs in studied area polluted slightly compared with other area. PAHs contents outside the cave and near cave gate were much higher than those within the cave with a little concentration change. In outside cave soils, the heavy molecular weight PAH (HMW) concentrations were higher than those of the light molecular weight PAH (LMW) (mean value 69.25 ng x g(-1) > 38.81 ngx g(-1)) while it was reverse inside cave (mean value 3.93 ng x g(-1) < 7.41 ng x g( 1)),which indicated the inner PAHs might came from atmosphere introduction and rainwater filtering from the top of the cave too. PAHs concentration distribution in the soils presented "trap effect" near the cave gate and a concentration superposition from the slope in the north cave wicket. HMW concentration near the cave gate was even larger by three times than that outside cave gate and a positive relation was found between PAH molecular weight and its falling tendence (R2 = 0.49). Deducing from the analysed data, PAHs concentration contributed by the cave gate was about 17 times of that of the slope. Temperature and molecular weight diversity were important factors influencing PAH concentration distribution. PMID- 19558085 TI - [Temperature sensitivity of wheat plant respiration and soil respiration influenced by increased UV-B radiation from elongation to flowering periods]. AB - Field experiment was carried out in the spring of 2008 in order to investigate the effects of increased UV-B radiation on the temperature sensitivity of wheat plant respiration and soil respiration from elongation to flowering periods. Static chamber-gas chromatography method was used to measure ecosystem respiration and soil respiration under 20% UV-B radiation increase and control. Environmental factors such as temperature and moisture were also measured. Results indicated that supplemental UV-B radiation inhibited the ecosystem respiration and soil respiration from wheat elongation to flowering periods, and the inhibition effect was more obvious for soil respiration than for ecosystem respiration. Ecosystem respiration rates, on daily average, were 9%, 9%, 3%, 16% and 30% higher for control than for UV-B treatment forthe five measurement days, while soil respiration rates were 99%, 93%, 106%, 38% and 10% higher for control than for UV-B treatment. The Q10s (temperature sensitivity coefficients) for plant respiration under control and UV-B treatments were 1.79 and 1.59, respectively, while the Q10s for soil respiration were 1.38 and 1.76, respectively. The Q10s for ecosystem respiration were 1.65 and 1.63 under CK and UV-B treatments, respectively. Supplemental UV-B radiation caused a lower Q10 for plant respiration and a higher Q10 for soil respiration, although no significant effect of supplemental UV-B radiation on the Q10 for ecosystem respiration was found. PMID- 19558087 TI - [Distribution of polycyclic aromatic hydrocarbons in soil profiles in southeast suburb of Beijing wastewater irrigation area]. AB - 3 borehole profiles samples were collected using Eijkelkamp soil sampler from the wastewater irrigation area of Beijing Southeast suburb. The soil samples were collected from surface to 5.5 m underground every 0.5 m. Physical-chemical properties of the samples such as clay content, total amount of clay minerals, cation exchange capacity (CEC), total organic compounds (TOC), etc. were analyzed. 16 polycyclic aromatic hydrocarbons (PAHs) defined by the U.S. EPA were also analyzed with gas chromatography-mass spectrometry (GC-MS). Results show that 14 PAHs were detected in the surface soil samples. The concentrations of PAHs range from 4 microg/kg to 428 microg/kg. Under the surface,both species and concentrations of PAHs in soil samples drop very fast. Only 2 rings and 3 rings PAHs were detected, which were naphthalene,phenanthrene, fluorene, acenaphthylene, acenaphthene, and fluoranthene. Clay content, total amount of clay minerals, CEC and TOC have significant correlations at 0.05 level between each other. Under the surface, clay contents also have a relationship with low rings PAHs concentration. Where the layers have higher clay contents,the PAHs have higher concentrations. The contents of PAHs changing along the profiles also show that, low-rings PAHs are easier to migrate than high-rings PAHs. The sequence of the migration capabilities of low-ring PAHs is acenaphthene > fluorene > naphthalene > phenanthrene > acenaphthylene > fluoranthene. However, high-ring PAHs were only detected in surface soil samples. That means low-rings PAHs can reach the deep layers of the vertical profiles under long-term wastewater irrigation. Therefore, the shallow groundwater has the possibility to be contaminated. PMID- 19558088 TI - [Effect of long-term fertilization on soil plough layer carbon storage in a reddish paddy soil]. AB - With a reddish paddy soil as test materials, soil profile nitrogen storage in long-term different fertilization system (1990-2006) have been investigated. The result indicated that recycling of organic matter significantly increased the soil profile C storage (ranged from 57.7 to 66.2 t/hm2), and it was increased by 18.7-27.2 t/hm2 compared with the soil profile C storage of 1990. But it was increased by 5.4 t/hm2 with only application of chemical fertilizers. Saturated carbon storage of paddy soils was 84.0 t/hm2, and the C sequestration potential ranged from 17.8-43.7 t/hm2 compared with the current soil profile carbon storage. The result showed that there was a significant relationship between soil bulk density and depth changes of profile soil. The organic C storage would be greatly underestimated by 20.6% or 11.3% if we only take 20 cm or 23 cm as the standard depth in the estimating method for organic C storage, it also would be underestimated the difference of treatments. The combined application of chemical fertilizer and organic matter is optimal for agricultural field based onsoil organic C storage and the carbon sequestration potential. PMID- 19558089 TI - [Mechanisms of the removal and remediation of phenanthrene and pyrene in soil by Pogonatherum paniceum]. AB - The mechanisms of the removal and accumulation of phenanthrene (PHE) and pyrene (PYR) by rock plant Pogonatherum paniceum were studied by pot experiments. Results showed that P. paniceum might effectively remove PHE and PYR from soils at their initial concentrations of 20 to 322 mg x kg(-1). About 50.97%-86.77% of PHE or 46.45%-76.7% of PYR was removed from the soils respectively after 70-day plantation of P. paniceum; the average removal rates of PHE or PYR from the soils by P. paniceum were 63.56% and 58.6% higher than those of CK1 (with addition of 0.1% NaN3), and 46.09% and 42.92% higher than those of CK2 (without NaN3). P. paniceum did show ability to accumulate polycyclic aromatic hydrocarbons (PAHs) from the soils; the contents of PAHs in its root and shoot increased withthe increase of PAHs concentrations in the soils. The bioconcentration factors (BCFs) for PAHs tended to decrease with increasing concentrations of these contaminants in soil. BCFs for PYR were higher than those for PHE, and BCFs for PHE (0.12 1.63) and PYR (1.03-5.61) in shoots were much lower than those in roots (0.21 3.08 and 1.31-10.11) at the same treatment. The mechanisms of phytoremediation processes by P. paniceum involve both biotic and abiotic factors. Contributions of each factor to the phytoremediation processes were estimated, which indicated that abiotic loss, plant accumulation, phytodegredation and microbial degradation accounted for 5.1%, 0.32%, 4.22% and 17.47% of the total removal of PHE, and 2.56%, 4.27%, 2.01% and 15.68% of PYR from soils, respectively. In contrast, 41.56% removal of PHE and 36.64% of PYR were attributed to plant-microbial interactions. Thus plant-microbial interactions are the main mechanisms for the remediation of soil PAHs pollution. PMID- 19558090 TI - [Effects of antimicrobial drugs on soil microbial respiration]. AB - The effects on soil microbial respiration of sulfonamides, tetracyclines, macrolides and so on were studied using the direct absorption method. The results show sulfamethazine, sulfamethoxazole, chlortetracycline, tetracycline, tylosin and trimethoprim inhibit soil respiration 34.33%, 34.43%, 2.71%, 3.08%, 7.13%, 38.08% respectively. Sulfamethoxazole and trimethoprim have the highest inhibition rates among all the antibiotics. In early incubation period (0-2 d), the concentrations above 10 mg x kg(-1) of sulfamethazine, sulfamethoxazole and trimethoprim remarkably decrease soil CO2 emission. The effects of these antibiotics vary with their concentrations too. Sulfamethoxazole and trimethoprim show good dose-response relationships. According to the standard of pesticide safety evaluation protocol, the six antibiotics pose a little risk to soil microbial environment. PMID- 19558091 TI - [Impact of phthalic acid easters on diversity of microbial community in soil]. AB - Phthalic acid easters (PAEs) are one of the most extensive organic and poisonous pollutants. BIOLOG and ARDRA method were used to analyze the impact of PAEs on microbial diversity in soil. BIOLOG analysis shows that the increase of average well color development (AWCD) is different in soils dealt with different concentrations of PAEs. With a higher PAEs concentration, the AWCD increases more slowly and the metabolic activity decreases more quickly. Physiological metabolism activity and carbon metabolic group show obvious difference in different samples. Although metabolic groups of carbohydrates and carboxylic acids were the dominant groups in control sample and low concentration sample, the dominant group is main polyamines in middle and high concentration samples. Principal component analysis (PCA) shows the differences of carbon metabolism. Four samples could be separated by PC1. Substrates of high positive correlation coefficients with PC1 were L-asparagine, 4-hydroxy benzoic acid and D-malic acid, but D-galacturonic acid, i-erythritol, gamma-hydroxybutyric acid and glucose-1 phosphate showed high negative correlation. ARDRA analysis showed that the diversity index decreased, with the increased concentration of PAEs. In a short time, PAEs could increase the diversity of microbial genotype in soil. PMID- 19558092 TI - [Effect of abamectin insecticide on the microbial community in broccoli phyllosphere]. AB - The indigenous microbial communities within the plant phyllosphere are highly diverse and include many different species of bacteria, filamentous fungi, yeasts and algae which play important ecological roles. This study was the first attempt to assess the impact of abamectin treatments on microbial communities of broccoli phyllosphere using two culture-independent techniques of phospholipid fatty acid analysis (PLFA) and terminal restriction fragment length polymorphism (T-RFLP). Results showed that low concentration of abamectin treatments did not affect the microbial biomass and microbial community structure of broccoli phyllosphere significantly. However, high concentration of abamectin treatments significantly change the microbial community structure including a decrease of total and bacterial biomass, and a decrease in the ratio of Gram-positive bacteria to Gram negative bacteria, but did not change the fungal biomass. Moreover, PLFA suggested that the number of unsaturated and cyclopropane phospholipid fatty acids (PLFAs: 16: 1omega9t, 18: 1omega7, cy17:0 and cy19:0) increased with high concentration abamectin treatment, while the saturated PLFAs i15:0, a15:0, i16:0 and a17:0 decreased. The appearance terminal restriction fragments (T-RFs: 58, 96, 236 and 420 bp) indicated that some bacteria might play a significant role in abamectin degradation in broccoli phylosphere, while the disappeared T-RFs (51, 89, 99, 338, 66, 223 and 482 bp) implied some other bacteria might potentially serve as microbial indicator of abamectin exposure. PMID- 19558093 TI - [Isolation of an anthracene-degrading strain Aspergillus fumigatus A10 and its degradation characteristics]. AB - An anthracene-degrading strain (A10) was isolated from contaminated environment and identified as Aspergillus fumigatus. The experimental results showed that the biodegradation rate of anthracene increased with the increasing time. Between 12 84 h interval, the biodegradation performed rapidly, while after this, the increase of biodegradation rate tended to become slow, and ultimately the biodegradation rate could achieve approximately 83%. The degradatinn rate of anthracene reached 79.37% within 5 days when the initial concentration of anthracene in mineral salts medium (MSM) was 10 mg/L, the inoculum dosage was 50 g/L (wet weight) and the cell age was 36 h. The concentration of anthracene had notable influence on degradation function of strain A10 and the highest degradation rate (92.17%) was achieved when anthracene concentration was 5 mg/L. The degradation rate could maintain about 60% with initial pH of MSM in the range of 5.0-7.5, and also, the anthracene could be better broken down when the temperature was 30 degrees C and dissolved oxygen was 4.30 mg/L. Certain amount of nutrition salts promoted the biodegradation of anthracene to some extent. Addition of lactose as co-metabolic substrate most favorably accelerated degradation of anthracene by about 37.15%. The mechanism research revealed that the biodegradation by strain A10 was a dynamic process in which extracellular sorption and intracellular degradation were included. FT-IR analysis exhibited that the structure of anthracene changed with the action of microbe, generating a series of metabolites, such as aromatic acid, aromatic ketone, aromatic aldehyde with one or two benzene rings, as well as saturated hydrocarbons. PMID- 19558094 TI - [Effects of typical organic pollutants on marine dioxide carbon system in simulated experiments]. AB - The effects of typical organic pollutants (ethanol, acetone, ON, OP) on pH and marine carbon dioxide system were investigated in laboratory simulated experiments. The results indicated that the growth of biological carbon fixed system was promoted by ethanol and acetone of low concentration (< 0.5 mmol x L( 1)) (t = 7 d). So pH increased while DIC, HCO3(-) and p(CO2) significantly decreased compared to the controls (p <0.01). The concentrations of ethanol and acetone which resulted in the maximum decrease of carbon dioxide parameters were 0.25 mmol x L(-1) and 0.1 mmol x L(-1), respectively. When the infusion of organic pollutants were higher than the "critical concentration", the carbon dioxide parameters increased and pH decreased compared to the controls with the decrease of carbon fixation strength of alga. As for ON and OP groups, when they were in the range of experimental concentration, decreasing trends of inorganic carbon in sea water were always found. The concentration changes of DIC (DeltaDIC) under the press of the four typical organic pollutants were significantly negatively correlated with the changes of dry weight of alga (p < 0.01 or p < 0.05) and the coefficient parameters were -0.902, -0.945, -0.898 and 0.918, respectively (t = 7 d). The main controlling factor to the variation of carbon dioxided system was the adaption of biological carbon fixed system to different concentration and kind of organic pollutants. PMID- 19558095 TI - [Toxicity evaluation of sewage treatment plant effluent of chemical industrial park along the Yangtze River on rat testicular germ cells in vitro]. AB - By using rat testicular germ cells in vitro toxicity testing method based on original cells culture, the reproduction toxicity of sewage treatment plant effluent of Chemical Industrial Park along the Yangtze River was evaluated, through cells changes in morphologic, activity and viability parameters. The results showed that both of the effluents from new developed Chemical Industrial Park A and provincial Chemical Industrial Park B contain reproductive toxic substances. The toxicity of Park A has more significant undergone changes in cells activity of sertoli cells (p < 0.01), spermatogenic cells (p < 0.05) and leyding cells (p < 0.05), lactate dehydrogenase activity (p < 0.01) and testosterone secretion (p < 0.01) than that of Park B. Sepermatogenic cells are more sensitive in indicating reproduction toxicity for testicular, compared with leyding cells and sertoli cells. This study demonstrated that, as an indispensable and complementary tool for water quality assessment, rat testicular germ cells in vitro toxicity testing based on original cells culture can be used to comprehensively evaluate the reproduction toxicity of sewage treatment plant effluent, and provide prompt and useful discharge quality information. PMID- 19558096 TI - [Quantitative retrieval of chlorophyll a concentration in Taihu Lake using machine learning methods]. AB - We evaluated the performance of two machine learning methods, artificial neural net (ANN) and support vector machine (SVM), for estimation of chlorophyll a in Taihu Lake from remote sensing data. The theoretical analysis has been done from basic theory and learning target of these two methods first. Then two empirical algorithms have been developed to relate reflectance of MODIS to in situ concentrations of chlorophyll a. The performance of ANN and SVM is comparatively analyzed in terms of validation, stability and robustness assessment and chlorophyll a distribution of Taihu Lake from two algorithms. The root of mean square deviation (RMSE) and average relative error (ARE) of validation data is only 5.85 and 26.5% of SVM retrieval model, however, RMSE and ARE of ANN model is 13.04 and 46.8%. Stability and robustness assessment suggest that SVM provides the better performance than ANN. And the retrieval results show that the chlorophyll a distribution of the whole lake from two algorithms is similar, however, the chlorophyll a concentration in the eastern region and central region of Taihu Lake is distorted by ANN model because of the limitations, such as learning target setting and over-learning in net construction. PMID- 19558097 TI - [Variation of spectrum of photosynthesis available radiation absorbed by underwater optical medium with depth in Lake Taihu in summer]. AB - Normalized spectrum of photosynthesis available radiation (PAR), which was absorbed by all kinds of medium per unit volume of water at different depths, was calculated by the data from downward and upward irradiance and absorption coefficients of phytoplankton, non-algae particles and chromophoric dissolved organic matter (CDOM) in 41 locations in Lake Taihu from July 29 to August 1, 2006. The results showed that there were two peaks at about 450 nm and 675 nm in the normalized spectrum of PAR absorbed by phytoplankton per unit volume of water, respectively. With the increasing of the depth, the peak at about 450 nm gradually weakened, and red-shift appeared from 450 nm. The above phenomena were much more obvious in the algae-type zone and the center of Lake Taihu. The PAR absorbed by phytoplankton per unit volume of water was shift to 600-700 nm which mainly originated from 400-500 nm to 600-700 nm. The transitional speed was slower in the grass-type zone but it was faster in Meiliang Bay(algae-type zone)and the center of Lake Taihu. In the above water, the PAR absorbed by non algae particles per unit volume of water mainly originated from 400-500 nm, but energy source gradually shifted to 500-600 nm with the increasing of depth. The variable characteristics of normalized spectrum of PAR absorbed by CDOM per unit volume of water were similar to that of non-algae particles. However, the advantage in grass-type zone which CDOM per unit volume of water absorbed PAR between 500 and 600 nm was less significant than that of non-algae particles per unit volume of water. PMID- 19558098 TI - [Phosphorus forms and its distribution characteristics in sediments and soils of water-level-fluctuating zone of the backwater reach from input river of Three Gorges Reservoir]. AB - The contents of different phosphorus forms in surface sediments and the soils of water-level-fluctuating zone of the backwater reach from input river of Three Gorges Reservoirwere measured, using thestandard measurement and test (SMT) procedure of phosphorus forms in freshwater sediments. Correlation coefficients between phosphorus forms and sample geochemical characteristics, such as organic matter and major components were analyzed. The results indicated that the TP content in surface sediments varied from 483.4-848.4 mg/kg, and the average content was 569.0 mg/kg. The TP content in surface sediments placed in the mid lower level, comparing with the TP content in surface sediment from the shallow lakes in the middle and lower reaches of Yangtze River. The TP content in soils of water-level-fluctuating zone varied from 488.9-1487.7 mg/kg, and the average content was 813.3 mg/kg. The average content of TP in soils of water-level fluctuating zone whole exceeded the surface sediment, indicating of the influence of human's activity on phosphorus contents in soils of water-level-fluctuating zone. Distribution characteristics of phosphorus forms were different between the sediment and soils of water-level-fluctuating zone: (1) The ratio of IP/TP in the surface sediments (average value 55.7%) was higher than that in soils of water level-fluctuating zone (average value 49.4%); (2) The domination phosphorus forms of IP in surface sediments was Ca-P (average ratio 83.5%), and the ratio of (Fe/Al-P)/IP was only 15%. The average ratio of Ca-P/IP in the soils of water level-fluctuating zone was 73.9%, whereas the ratio of Fe/Al-P in IP raised to 22%; (3) The average content of active phosphorus (OP + Fe/Al-P) in surface sediment was 261.8 mg/kg, the average ratio of active phosphorus in TP was 49%, whereas the average content of active phosphorus (OP + Fe/Al-P) in soils of water level-fluctuating zone was 405.7 mg/kg, the average ratio was 54%. The active phosphorus in soils of water-level-fluctuating zone would become potential pollution sources of water under correct environmental conditions,therefore, the potential impact of phosphorus on water eutrophication in soils of water-level fluctuating zone couldn't be neglected. PMID- 19558099 TI - [Distribution of phosphorus species in the sediment from Weishanhu district in Lake Nansihu]. AB - In order to learn the phosphorus (P) species distribution in sediments of Lake Nansihu, 0-25 cm sediments from Weishanhu district in Lake Nansihu were analyzed with a sequential extraction method. The results show that: the average values of Ex-P, Al-P, Fe-P, Oc-P, Ca-P, De-P, Org-P in Weishanhu district sediments are 5.62 mg/kg, 4.08 mg/kg, 12.25 mg/kg, 13.34 mg/kg, 116.67 mg/kg, 232.36 mg/kg and 396.79 mg/kg respectively, and the rank order of P-fractionation for Lake Nansihu is Al-P < Ex-P < Fe-P < Oc-P < Ca-P < De-P < Org-P. The vertical phosphorous species distribution exhibits the obvious rule that exchangeable P (Ex-P), Fe bound P (Fe-P), occluded P (Oc-P), organic P (Org-P) value decrease with depth, while the values of Al-bound P (Al-P), authigenic calcium bound P (A Ca-P), detrital apatite P (De-P) increase. The Sum1 content (the sum of Ex-P, Al-P and Fe-P) in surficial sediments is remarkably positively correlated with the PO4(3-) concentration of overlaying water, to which Fe-P content contributes the most with the correlation index 0.72. In spatial phosphorous species distribution, the discrepancy of potential active species(Ex-P, Al-P and Fe-P) is greater than those inert ones (Oc-P, Ca-P and De-P), and Org-P is only second to those potential active species. PMID- 19558100 TI - [Phosphorus pollution of the sediment from Hangzhou urban river and hydraulic simulation of phosphorus release]. AB - The phosphorus pollution of the sediment from Dongxin River, an urban river in Hangzhou, was surveyed. Phosphorus release under various flow velocity (0.002 0.02 m x s(-1)), quality of overlying water (the total phosphorus concentration: 0.18, 0.072 and 0.007 mg x L(-1)), depth of overlying water (5, 10 cm) were carefully studied in a hydraulic simulation setup. The results indicate that phosphorus was feasible to release from the sediment with high ratio of water content (40.09%-68.57%) under alkali condition (pH 7.2-8.3). The phosphorus release was also influenced by high content of phosphorus (1.04-2.51 g x kg(-1)) in the sediment. Phosphorus release was mainly in the form of suspended solid under dynamic hydraulic condition, and depended definitely on flow velocity. The release rate reached 147.36 g x (m2 x h)(-1), which approached the maximum value, at flow velocity 0.008 m x s(-1) (i.e. 0.05 m x s(-1) in archetype). Phosphorus release, especially the initial release rate, was obviously affected by the quality of overlying water. Phosphorus tended to release under river water than tap water. However, the depth of overlying water had little influence on phosphorus release in shallow water. Therefore, it is possible to restrain or enhance phosphorus release by adjusting the hydraulic conditions and the quality of overlying water. PMID- 19558101 TI - [Transformation of inorganic nitrogen in slag-wetland during the start-up period]. AB - Lab-scale subsurface flow slag-wetlands were constructed to study the removal efficiency and transformation processes of low-concentration inorganic N during the start-up period. As for ammonium-dominated wastewater, the removal rates of total nitrogen (TN), ammonium nitrogen (NH4(+)-N), nitrate nitrogen (NO3(-)-N) and nitrite nitrogen (NO2(-)-N) were 0.12 g x (m2 x d)(-1), 0.07 g x (m2 x d)( 1), 0.10 g x (m2 x d)(-1) and 0.04 g x (m2 x d)(-1), respectively. Nitrifying bacteria was not detected by fluorescence in situ hybridization (FISH) on the slag. NH3 volatilization is the main contribution for N removal resulting from high pH (> 10) and surface soil absorbed most of NH3. For nitrate-dominated wastewater, the removal rates of TN and NO3(-)-N were 0.23 g x (m2 x d)(-1) and 0.48 g x (m2 x d)(-1), and NO2(-)-N accumulated by 0.22 g x (m2 x d)(-1) during the process of denitrification. Removal efficiency of inorganic N for nitrate dominated wastewater was higher than that for ammonium-dominated wastewater during the start-up period of slag-wetlands, so steel slag can be used as a substrate in constructed wetlands for extensive treatment of nitrate pollution. PMID- 19558102 TI - [Survey of estrogenic activity of Songhua River water and drinking water of Harbin]. AB - Solid phase extraction/yeast two-hybrid system was used to investigate estrogenic effects of Songhua River water of Harbin reach and drinking water of Harbin. Results show that the estrogenic activities of Songhua River water in summer and autumn are 528-965 pg x L(-1) estradiol equivalents (EEQs), and the estrogenic activity of source water is in a high level. The removal rates of estrogenic activity are 34.6%-50.5% in a waterworks of Harbin. It reveals that estrogenic pollutants in source water are not efficiently removed in the plant. As a result, a level of 298-718 pg x L(-1) of estrogenic activity is left in the effluent of the plant. Results also demonstrate that coagulation-sedimentation process is the most stable treatment in the removal of estrogenic activity among the three water treatment processes applied in the plant, with a removal efficiency of 11.4% 26.1% which accounts for 32.9%-69.0% of the total removal. Processes of sand filtration and chlorination are not stable, and estrogenic activities of their effluents sometimes are higher than those in their influents. Moreover, the estrogenic activity of tap water is increased to the maximum of 44.9% in comparison with that of the effluent of the plant due to the secondary contamination produced in the process of pipe delivering. The estrogenic activities of tap water of Harbin are 347-1,362 pg x L(-1) measured in seven sampling events, with a mean value of 738 pg x L(-1). PMID- 19558103 TI - [Rapid detection of rotavirus in water samples using immunomagnetic separation combined with real time PCR]. AB - A quantitative and rapid detection method for rotavirus in water samples was developed, by using immunomagnetic separation combined with reverse transcription and real time polymerase chain reaction (IMS-RT-real time PCR). Magnetic beads coated with antibodies directed against group A rotavirus were used to capture and purify the virus in water samples. The experimental results showed that IMS was optimized when 1 mL samples were supplemented with 10 microL of immunomagnetic beads, 2.5 microL of Tween 20 and incubated for 2 h. The IMS method was employed in the detection of rotavirus in seeded virus eluant such as 3% beef extract successfully and thus manifested its compatibility with established virus concentration methods. The IMS-RT-real time PCR method could yield quantitative results within about 5 h with a detection limit at 1 x 10(4) copies/mL (equivalent to 3-4 PFU/mL). The method exhibited a high level correlation (R2 = 0.9816) with cell culture assay, indicating that it could perform as well as cell culture assay does in infection tests. And the method functioned satisfactorily in seeded concentrate of secondary waste water treatment plant effluent, reclaimed water, surface water and tap water. PMID- 19558104 TI - [Factors affecting formation of THMs during dissolved organic nitrogen acetamide chlorination in drinking water]. AB - Chlorination disinfection greatly reduced bacteria and virus in drinking water. However, there is an unintended consequence of disinfection, the generation of chemical disinfection by-products (DBPs). Dissolved organic nitrogen (DON) as the important precursor of DBPs is of current concern. As acetamide (AcAm) occur in important bimolecular, we studied formation pathways for THMs during chlorination of model AcAm. The experiments are designed by Plackett-Burman and Box-Behnken methods. Factors affecting formation of THMs such as AcAm initial concentration, chlorine dosage, pH, temperature, Br(-) concentration and contact time were investigated. The results indicate that AcAm initial concentration, pH and temperature have little effects on formation of THMs. On the contrary, three other factors have important effects on formation of THMs, especially Br(-) concentration. The capacity of THMs generation varies very little when Br(-) has a constant concentration. Generation amount of THMs attach maximum under the condition that dosage of active chlorine, Br(-) concentration and contact time is 8.77 mg/L, 0.77 mg/L and 6.20 h respectively. Bromine ion plays a catalysis role on THMs formation. Controlling the concentration of bromine ion can reduce total generation amount of THMs via AcAm. Bromine partition coefficient tends to increasing along with contact time lapse. Controlling chlorination reaction time can lower the cancer risk. At last, the pathway is proposed for THMs formation via AcAm, and the catalysis mechanism of Br(-) was addressed. PMID- 19558105 TI - [Oxidization characteristic of chlorine on the biofilm in simulated drinking water distribution system]. AB - In the article, the inactivation effect of chlorine on E. coli biofilm and the influence of chlorine oxidization on the contents of assimilable organic carbon (AOC), microbially available phosphorus (MAP) and bacterial regrowth potential (BRP) was investigated in the simulated drinking water distribution system. Results showed that chlorine resulted in more efficient reduction on suspended E. coli than did in biofilm. The inactivation effect of E. coli was influenced by chlorine concentration. Likewise, higher chlorine concentration resulted in more E. coli inactivation rate at the same CT (chlorine concentration multiply by time) value, when biofilm was oxidized by chlorine. Concentrations of AOC and MAP in bulk water increased owing to organic substance dissolved from biofilm. The AOC concentration increased from 20.78 microg/L to 120.17 microg/L, and the MAP was increased from 0.11 microg/L to 0.17 pg/L, and the Chlorine oxidization enhanced BRP concentration in the bulk water. BRP reached maximum at 1.10 x 10(7) CFU/mL when chlorine concentration was 1.0 mg/L, CT value was 100 mg x min/L. PMID- 19558106 TI - [Influence factors of bromate formation in Yangtze River water during ozonation process]. AB - A serial bench-scale experiments were conducted to study the effect of ammonia, pH, initial dissolved ozone and temperature on bromate formation during ozonation process in Yangtze river water, where bromide anion (Br(-)) was detected. Ammonia has little suppression effects on bromate production in Yangtze water at pH 7.35 and 0.6 mg/L ammonia background concentration, for it can't block the indirect direct path. Within our experiment's conditions, at the initial dissolved ozone from 0.8 mg/L to 2.5 mg/L, it should be kept below 1.2 mg/L in order to avoid bromate standard violation. BrO3(-) formation is sensitive to pH variation. BrO3( ) production is 8 microg/L when pH decreases to 6.3. Lower temperature leads to both slower ozone decomposition and lower production of bromate. BrO3(-) production is 9.1 microg/L at 10 degrees C, 100 microg/L initial Br(-) and 1.6 mg/L initial dissolved ozone. PMID- 19558107 TI - [Effect of preozonation on microfiltration for Huangpu River]. AB - This study investigated the effect of preozonation on organic matter and microfiltration membrane filtration, Huangpu River surface water ozonated was filtrated by MF membrane. The results show that highest rejection efficiency of DOC and UVr by ozone are 10% and 71% respectively during 0.5-3.0 mg/L ozone dosage. With increasing of ozone concentration, organic substances of 2 x 10(3)-7 x 10(3) reduce gradually, while organic matter of which less than 0.5 x 10(3) change slightly. Ozone oxidizes more hydrophobic fraction to hydrophilic one and only few to inorganic matter. Changes of organic matter composition affect membrane filtration resistance, thereby improve membrane flux. There is the optimal dosage with ozone of 1.5 mg/L made membrane flux maximum in the range of 0.5-3.0 mg/L ozone dosage. The reason is change of organic matter composition which made membrane filtration resistance minimum in the ozone dosage of 1.5 mg/L. PMID- 19558109 TI - [Biological reduction of perchlorate and optimization]. AB - The potential of bioremediation to treat perchlorate-contaminated water by acclimating anaerobic activated sludge was investigated in this study. Factors such as carbon source, initial pH, temperature, domesticated sludge concentration and dissolved oxygen affecting the removal of perchlorate were studied in detail by shaking-bottle point tests using acetate as carbon source. Results showed that 50 mg/L perchlorate was completely reduced by 1.0 g of domesticated sludge added with 1.2 g/L of acetate under the conditions of 35 degrees C and initial pH 8.0. Dissolved oxygen in the system would restrain the reduction of perchlorate. In addition, column bioreactor was used to treat perchlorate continuously, with the minimum residence time of 6 h. PMID- 19558108 TI - [Removal characters of ozone-biological activated carbon process for typical pollutants in southern brooky regions of China]. AB - The products of relative molecular weight (Mr) distribution, bromate (BrO3(-)) and trihalomethanes (THMs) were studied by ozone-biological activated carbon (O3 BAC) process for treating organic matters and bromide (Br(-)) in water source of southern brooky regions of China. The experimental results showed that dissolved organic matters (DOC) with Mr lower than 10(3) accounted for 80% of the total. The removal rate of DOC and SUVA (UV254/DOC) were 8% and 14% respectively by traditional treatment process with main removalonly for ones with Mr higher than 100 x 10(3). Only 30% of DOC and 31% of SUVA were decreased by O3-BAC process for the removal of ones with Mr between 10(3) and 5 x 10(3), in which the biotic degradation was certainly restricted by predominant organic matters of hydrophilic and Mr was lower than 1000. An obvious increase of BrO3(-) occurred in the effluent from ozone oxidation process when the dose of ozone beyond 2 mg/L which increased Br(-) concentration. This could increase the product of BrO3(-). A poor and unstable removal effect of BrO3(-) was observed in the effluent of BAC process during the experiment. Each species of THMs, decreasing 40% of total, was reduced by O3-BAC treatment compared with the traditional treatment process. But the products of brominated trihalomethanes, especially CHBr3 would be markedly increased by enhanced chlorine dosage and Br(-) concentration. PMID- 19558110 TI - [Preparation of a novel graphite electrode and its effect on phenol degradation]. AB - In order to find an effective, stable, environmentally compatible electrode for the elimination of organic pollutants, preparation of a novel graphite electrode (NGE) was explored using graphite, epoxy resin, curing agent and acetone as materials. Water, ethanol aqueous solution, acetone, ultrasound-acetone and electrochemical method were used to pretreat the electrode. The first three methods obtained poor effect. Ultrasound-acetone could improve the pretreatment, but satisfied results were difficult to achieve. Electrochemical method could work effectively. Electrochemical oxidation of phenol was carried out on NGE and commercial graphite electrode (CGE), respectively. UV spectroscopy was used to study evolution of phenol oxidation. The results showed that CGE resulted in accumulation of bensoquine in the electrolyte, though it was more effective than NGE on phenol oxidation. While much less benzoquinone was found in the electrolyte for NGE. TOC removal rate were 40% and 31% for NGE and CGE, respectively. SEM study showed that CGE was severely corroded after phenol degradation, while NGE was undamaged, showing good stability. PMID- 19558111 TI - [Nitrate removal from wastewater using rice straw as carbon source and biofilm carrier]. AB - In this research,agriculture waste-rice straw was investigated as the sole carbon source as well as biofilm carrier to remove nitrate from wastewater in up-flow laboratory reactors. The experimental results indicated that the startup of the system was quick and a high nitrate removal efficiency was observed. The accumulation of nitrite was not observed in this process. Moreover, high nitrate concentration in the influent had a significant effect on nitrate removal efficiency. The reactor was able to accommodate a wide range of pH (6.5-8.5) and DO (1.0-3.5 mg/L). A time-dependent decrease in nitrate removal efficiency was observed after 84 days of operation. The results showed that rice straw could be used as an economical and effective carbon source for denitrification. PMID- 19558112 TI - [Simultaneous removal of nitrate and 4-chlorophenol from contaminated water using solid-phase denitrification process]. AB - A novel biodenitrification process, termed as "solid-phase denitrification", was used to remove nitrate and 4-chlorophenol (4-CP) from water. The experimental results indicate that nitrate can be effectively removed using biodegradable meal box (BMB) as carbon source and biofilm carrier of denitrifying microorganisms. Volumetric denitrification rate was 24.0 mg/(L x h) with the 50 mg/L of initial NO3(-)-N concentration under the batch experiments. Denitrification was facilitated with 4-CP concentration which was less than 30 mg/L, and was repressed with over 40 mg/L of 4-CP. When the initial concentration of 4-CP was 5 mg/L and 30 mg/L, the removal efficiency of 4-CP was 90% and 71% within 8 h under denitrifying condition, respectively. 4-CP removal was due to both the absorption onto BMB and degradation by microorganisms attached to BMB. PMID- 19558113 TI - [Efficiency of atrazine degradation by O3/H2O2]. AB - The endocrine disrupter Atrazine was oxidized by O3/H2O2 system and the products were analyzed to assess the degradation efficiency of Atrazine. When it's initial content was 2 mg/L and O3 dosage was 7.5 mg/L, Atrazine was removed about 27.2% after 5 minutes. Under the same condition, H2O2/O3 molar ratio was 0.75, Atrazine maximum removal rate reached 96.5%, which suggested that Atrazine could be degraded by O3/H2O2 system effectively. Ion Chromatography (IC) analysis showed that concentrations of chloride and nitrate ions were increasing along with the Atrazine content decreasing. Gas Chromatography-Mass spectrometry (GC-MS) and Liquid Chromatography-Mass spectrometry chromatograms (LC-MS) analyzing illuminated the existence of de-ethyl-atrazine, de-isopropyl-atrazine and de chloro-atrazine, which indicated the Atrazine could not be destroyed completely by O3/H2O2 system. Consequently, it should be combined with GAC (Granular Activated Carbon) or other techniques while used as primary treatment unit or emergency measure. PMID- 19558114 TI - [Competition and optimization of AOB and NOB for domestic wastewater treatment at normal temperatures]. AB - At normal temperature of 19 degrees C +/- 1 degree C and seed sludge from the system of conventional nitrogen removal via nitrate composed of both ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB), the competition and optimization of AOB and NOB was investigated in a lab-scale SBR treating domestic wastewater. The reactor was operated under the aeration rate of 40 L/h with the average DO concentration below 1.0 mg/L, and combined with the control of aeration time, i.e., aeration was stopped before the pH "ammonia valley" was detected. After AOB became pre-dominant nitrifying bacteria (nitrite accumulation rate NO2(-)-N/NOx(-)-N reached 50%), the aerobic durations were gradually prolonged from 3 h to 4 h and 5 h to improve the ammonia removal efficiencies and enhance AOB dominant. By the above operational pattern, nitrogen removal via nitrite was successfully achieved with NO2(-)-N/NOx(-)-N over 95%. Fluorescence in-situ hybridization (FISH) analysis showed that AOB was accounted for 9.97% of total biomass. The on-line control of aerobic duration was an effective method to maintain the partial nitrification and the NH4(+)-N removal efficiency was above 97%. The research results also proved that at normal temperatures, for the seed sludge from the conventional nitrogen removal via nitrate, AOB would not become dominant nitrifying bacteria only by the aeration time control without the DO restriction. PMID- 19558115 TI - [Half-nitrosofication of dewatering liquid of digested sludge with high ammonia]. AB - The affecting factors and controlling methods of how realizing and maintaining half-nitrosofication of dewatering liquid of digested sludge were investigated in an A/O process. The experimental results showed that the nitritation was obtained in 29 d under bulk liquid temperature of 9-20 degrees C, average DO of 5.4 mg/L, SRT of about 30 d and influent ammonia loading rate of 0.64 kg/(m3 x d). The nitritation with 70% nitrite accumulation efficiency was maintained during 65 d when average FA concentration in A/O reactor was more than 4 mg/L. Furthermore, the half-nitrosofication with effluent ammonia and nitrite ratio of 1:1.32 was obtained. When the ammonia loading rate reduced to 0.19 kg/(m3 x d) (FA < 1 mg/L), the nitritation disappeared, and the high FA as a main factor of maintaining half-nitrosofication was proved again with FISH under the condition of different FA. TN removal concentration was about 91 mg/L under influent COD of 282 mg/L and raw C/N ratio of 0.85 in the A/O system, which due to nitritation saving much carbon sources. The study showed that the half-nitrosofication of the dewatering liquid was obtained and stably maintained when FA was more than 4 mg/L in the A/O reactor by dynamic controlling influent ammonia loading rate and pH value in the system under middle or low temperature, high DO, long SRT. PMID- 19558116 TI - [Treatment of ANAMMOX technique on treating high concentration poultry wastewater]. AB - Taking the effluent of the UASB-shortcut nitrification technique of typical high concentration poultry wastewater as the object, it is discussed that the ANAMMOX is applicable as a treatment of denitrification. Firstly, it successfully starts ANAMMOX reactor by the denitrifying activated sludge, on this basis, finding the optimal ammonia loading is about 0.2 kg/(m3 x d) and the HRT of system is 2 d. Moreover, after studyingon the operating conditions, the optimum is that pH is about 7.50; temperature is 30 degrees C without adding organic carbon. Under optimum conditions, the removal efficiencies of ammonia and nitrite nitrogen could be above 85% and 95%, the system is running well and has reproducibility. Finally, it reaches that the reaction rates of ammonia and nitrite nitrogen are 0.0126 d(-1) and 0.0131 d(-1) through dynamic analysis. So it can make the theory basis for spreading and application of after/continuous process, neural network simulation and "UASB + SBR shortcut nitrification + ANAMMOX + land treating system" newly combined process. PMID- 19558117 TI - [Effect of organic loading rate on granular SBR]. AB - The effect of organic loading rate on granular SBR was investigated. When the organic loading rate gradually reduced from 1.8 kg/(m3 x d) to 1.575, 1.125 and 0.67 kg/(m3 x d), the characteristics of granular sludge changed greatly, including disintegration of granules, washout of sludge, reduce of granulation rate and settleability of granule, shift of granule shape from regular sphericity to different shapes of asteroid and bacilliform and the increase of color deepness. The granular SBR system reached COD and TP removal rates were about 90% and 70%, respectively, which were not influenced by the reduce of organic loading rate. However, the simultaneous nitrification and denitrification could be greatly influenced by organic loading rate reduce. When organic loading rate reduced from 1.8 kg/(m3 x d) to 0.67 kg/(m3 x d), the removal rates of ammonia and TN decreased 45% and 40%, respectively. PMID- 19558119 TI - [Functional microbial community in a membrane-aerated biofilm reactor operated for completely autotrophic nitrogen removal]. AB - The 16S rDNA-based molecular technique was applied to investigate the functional microbial community of a membrane-aerated biofilm bioreactor (MABR) that was used for completely autotrophic nitrogen removal over nitrite (CANON). The relationships among two kinds of key bacteria responsible for CANON: aerobic ammonia-oxidizing bacteria (AOB) and ANAMMOX bacteria, and their possible distributions in the MAB were discussed based on the microbial community analysis. FISH analysis showed the existence of two visible active layers in experimental MAB. One was the partial nitrifying layer located in the region of oxygen-rich membrane-biofilm interface, dominated by NSO190-positive AOB. The other was the ANAMMOX active layer located in the region of anoxic liquid-biofilm interface, dominated by PLA46 and AMX820-positive ANAMMOX microorganisms. As a result, the AOB as well as ANAMMOX bacteria were present and active in experimental MABR, and the cooperation between AOB and ANAMMOX bacteria was considered to be responsible for CANON. PMID- 19558118 TI - [Implement of completely autotrophic nitrogen removal in EGSB reactor and its operation optimization]. AB - An expanded granular sludge bed (EGSB) reactor inoculated simultaneously with aerobic ammonium oxidation sludge and anaerobic ammonium oxidation sludge were start-up to enrich completely autotrophic nitrogen removal granular sludge. Total nitrogen (TN) removal rate reached 0.101 kg x (m3 x d)(-1). Based on hypothesis of boundary layer, the transfer process between granular sludge and bulk liquid was modified, which was coupled with substance transfer process in granular sludge and aerobic ammonium oxidation, anaerobic ammonium oxidation, nitrite oxidation process, and completely autotrophic nitrogen removal model was found. The model was validated with the experimental results. According to simulation results, the operation of the reactor was optimized, TN removal efficiency and TN removal rate were increased from 52% to 61% and 0.103 kg x (m3 x d)(-1) to 0.114 kg x (m3 x d)(-1) respectively. PMID- 19558120 TI - [EPS, SMP and microbial biodiversity under the oligotrophic environment]. AB - In order to investigate the extracellular polymeric substances (EPS), soluble microbial product (SMP) and microbial biodiversity of the sludge lacking of nutrition, to provide evidence for optimizing MBR's operation and lessening membrane fouling, a series of experiments were undertaken under the oligotrophic environment using the sludge from the MBR of Tianjin University. The contents of EPS and SMP were determined, and microbial biodiversity was analyzed by PCR-DGGE technology and cloning-sequencing. Furthermore, the sequences were used for homology analysis and then the phylogenetic tree was constructed. In the initial stage, the EPS and SMP contents raised from 15.04 mg/g and 0 mg/g to 17.99 mg/g and 3.29 mg/g, respectively. Along with the experiment progress, the EPS content dropped to 2.40 mg/g; but the SMP content varied around 3.5 mg/g. This indicates that the EPS and SMP possess the buffer action towards the variation of the environment, and the microorganism can utilize the EPS and SMP under the condition of oligotrophic. Because of the utilization of the EPS and SMP, the Shannon index of the sludge raised from 0.81 to 1.09. Then, it began to decline and stabilized at 0.95 finally. The clone and sequence results reflect that the microbial structure is very rich, and most of the dominant species are uncultured bacterium. Some of the bacterium, which are mostly belong to Bacteroidetes, Flavobacterium, Saprospiraceae and Firmicutes, can degrade the EPS and SMP by secreting protein and polysaccharide hydrolyzing enzymes. PMID- 19558121 TI - [Characterisation of excess sludge reduction in an anoxic + oxic-settling anaerobic activated sludge process]. AB - An energy balance analysis method with auto calorimeter being adopted was introduced to determine calorific values of sludge samples in influent and effluent of uncoupling tank in an anoxic (A) + oxic-settling-anaerobic (OSA) process and a reference system. The affiliation of sludge amount change and its energy content were studied, as well as potential of excess sludge reduction was evaluated through modifying performance of uncoupling tank. The characteristi s and causes of sludge reduction in OSA system were deduced according to energy and matter balance analysis. Results show that when the hydraulic retention time (HRT) of uncoupling tank are 5.56 h, 7.14 h and 9 h, the excess sludge reduction of whole A + OS Asystem are 1.236 g/d, 0.771 g/d and 0.599 g/d respectively. Energy content of sludge flows into and out of the uncoupling tank changes, the specific calorific value of sludge in effluent is inclined to be higher than that in influent with the HRT of the tank increasing: there isn't any significant difference of sludge calorific values between influent and effluent at 5.56 h, while the differences are in 99-113 J/g at 7.14 h, and 191-329 J/g at 9 h. Sludge in uncoupling tank would decay and longer HRT will result in more attenuation. It could be concluded that excess sludge reduction of A + OSA system is caused by both of sludge decay in uncoupling tank and sludge proliferation in AO reaction zone. PMID- 19558122 TI - [Effect of 2-aminophenol and 3,3',4',5-tetrachlorosalicylanilide on biomass yield of activated sludge]. AB - The effects of 2-aminophenol and 3,3',4',5-tetrachlorosalicylanilide on biomass yields were studied. The results shows that when AP is 15 mg/L and TCS is 1.2 mg/L, observed growth yields (Yobs) of the sludge are reduced by 21% and 52% with low reduction of COD removal efficiency. Removal rates of ammonia decrease by 32% with TCS but are not affected obviously by AP. The effects of AP and TCS on dewater ability and settle ability of sludge were also investigated. The dewater ability of sludge is not affected by the two uncouplers. The SVI values of sludge with TCS increase slightly. According to microscope observation, the sludge microbe population changes. Meanwhile quantities and kinds of protozoan and metazoan were reduced. PMID- 19558123 TI - [Decoloration of reactive turquoise blue by acidified sludge-bentonite granule]. AB - Using sludge as pore-forming agent, bentonite granule was acidified by sulfuric acid solution as a decolorant. The specific surface area and SEM were performed to characterize the structure of samples, and the new acidified sludge-bentonite granule was applied to the decoloration of reactive turquoise blue. The influencing factors of pH value, dosage, reaction time and reaction temperature were studied on the removal of the dyes. The important thermodynamics parameters (DeltaH0, DeltaS0, DeltaG) and the activation energy Ea were also acquired by experiment data processing. The results indicated that the adsorption isotherm fitted the isothermal adsorption equations of Langmuir better than Freundlich. The adsorption dynamics followed the law of the pseudo-second order kinetic equation, while the adsorption rate is 313 K > 303 K > 293 K. The low value of Ea which is 5.52 kJ x mol(-1) shows that physical adsorption is primary. And DeltaH0 > TDeltaS0 means that the influence of enthalpy is more remarkable than the entropy in the activation reaction. DeltaG > 0 also means the chemical reactions are not spontaneous. PMID- 19558125 TI - [Relationship between typical organic matters in domestic wastewater and water characteristic parameters in activated sludge models]. AB - The contribution of typical organic matters such as proteins, sugars, lipids and linear alkylbenzene sulfonate (LAS) to COD of the domestic wastewater was investigated. Nitrate utilization rate was used to determine wastewater characteristic parameters. Relationship between these typical organic matters and wastewater characteristic parameters (Ss, Xs, SI, XI) in activated sludge models were investigated. The results showed that YH of activated sludge under denitrifying conditions was 0.683. Proteins, sugars, lipids and LAS accounted for 24%-35%, 17%-35%, 5.78%-10.56% and 3.77%-7.23% of the total COD, respectively. It indicated that these four pollutants were the main COD source in the domestic wastewater. SS and XI were in the ranges of 22%-29% and 29%-38% of the total COD, respectively. Concentrations of the four typical target organic matters (proteins, sugars, lipids and LAS) correlated well with the wastewater characteristic parameters (Ss, Xs, SI, XI) of activated sludge models (ASMs) with the correlative coefficients above 0.9. PMID- 19558124 TI - [Distribution character of synthetic musks in urban sewage sludges]. AB - The characteristics of occurrence and distribution of 8 synthetic musks in 15 sewage sludges in Shanghai were studied. The results indicated that HHCB and AHTN were the two main components in all sludges, with the mean concentration of 2.92 mg x kg(-1) (0.81-6.39 mg x kg(-1)) and 1.96 mg x kg(-1) (0.35-3.11 mg x kg(-1)), respectively; and the concentrations of ADBI, AHMI, MK were less. The distributions detected in sludges are in accordance with the usage patterns in China. Total concentrations of 8 synthetic musks were ranged from 1.16-9.57 mg x kg(-1), which were lower than the results in previous studies. Concentrations in sludges are influenced by ratio of domestic wastewater in influent, sewage load and sewage treatment methods. The good linear relationships among HHCB, AHTN and ADBI indicate that these components have the same source: domestic wastewater. The consumption rates of HHCB and AHTN connected to per inhabitant in Shanghai region were estimated, which are low compared with those found in European. The potential impacts on agricultural soil were also assessed. PMID- 19558126 TI - [Day-night variation of carbonaceous aerosols in PM10 during winter and spring over Xi'an]. AB - The purpose of this study was to investigate the day-night variation of carbonaceous aerosols in PM10 during spring and winter over Xi'an. PM10 samples were collected during 19 Dec 2006 to 21 Jan 2007 (Winter) and 1 Apr 2007 to 30 Apr 2007 (Spring). Organic carbon (OC) and elemental carbon (EC) concentrations were measured using thermal-optical method. PM10, OC, and EC concentrations in winter were 455.0, 62.4, and 7.5 microg/m3 during daytime, and 448.7, 66.1, and 6.9 microg/m3 for nighttime, respectively, while in spring were 397.9, 26.7, and 6.9 microg/m3 for daytime, and 362.1, 31.9, and 8.6 microg/m3 for nighttime. The correlation coefficient of OC and EC during daytime in winter was 0.44, while a strong relationship between OC and EC was observed in spring daytime, indicating that the emission sources of carbonaceous aerosol in winter were more complicated than those in spring. Due to high OC levels and the weaker dispersal ability of atmosphere, the mean concentrations of SOC during winter (8.9 and 10.2 microg/m3 at daytime and nighttime) were much higher than those in spring (2.8 and 3.4 microg/m3). Factor analysis on the eight carbon fraction indicated that coal combustion and biomass burning were the major sources for carbonaceous aerosol in winter, while vehicle exhaust played an important role in spring. PMID- 19558127 TI - [Investigations of microscopic morphology of individual inhalable particulates in Macao in summer]. AB - By means of high resolution field emission scanning electron microcopy (FESEM) and image analysis (IA), microscopic morphologies and the number-size and volume size distributions of different types of individual inhalable particles collected in Macao in summer of 2007 were investigated. The results showed that the soot aggregates and mineral particles were ubiquitous in the PM10 of Macao in July, 2007. The mineral particles mostly showed an irregular morphology and the soot aggregates displayed chain-like and fluffy morphology. The soot aggregates accounted for approximately 30%-40% in average by number, and the mineral particles accounted for approximately 20%. The relatively higher percentages of more soot aggregates implied that emission from motor vehicle exhausts was serious in Macao during summer. The number-size distributions revealed that the majority of the particles in PM10 of Macao were fine particles, with the equivalent diameter being concentrated in a range from 0.2-0.4 microm. A distinct spatial difference existed in the types of particles collected at different sampling sites. Soot particles dominated the busy transportation sites and the tunnel site, while the mineral particles had a higher proportion at the dock site. PMID- 19558128 TI - [Instantaneous emission simulation for light-duty diesel vehicle with different driving cycles by CMEM model]. AB - CMEM model for calculating time based instantaneous emission from light duty diesel vehicle and its input parameters were introduced. On-board test data were used to validate the simulation results. The relative error of THC, CO, and NOx are 14.2%, 3.7% and 32.7%, respectively, while the correlation coefficients reach 0.73, 0.72 and 0.87. The instantaneous emissions of the light duty diesel vehicle simulated by CMEM model are strongly coherent with the transient driving cycle in Shanghai. The simulation of instantaneous emissions and fuel economy under the ECE-15 cycle, FTP cycle, Japan 10-15 cycle and the cycle of shanghai arterial road show that the instantaneous emissions decline with the increase of the vehicle speed, especially from 0-10 km x h(-1) to 10-20 km x h(-1). The acceleration process dominated the whole emissions, which contributes over 30% of the total emission, and sometimes it even reaches over 70%. The contributions of shanghai arterial road for idle condition are 40% and 30%, emission factors of CO are 1.3, 1.5 and 1.4 times of ECE-15 cycle, FTP cycle, Japan 10-15 cycle respectively; THC are respectively 1.5, 2.1 and 1.9 times of above cycles; and emission factors of NOx are respectively 1.2, 1.3 and 1.3 times of ECE-15 cycle, FTP cycle and Japan 10-15 cycle. The fuel economy of the light-duty diesel car on shanghai arterial road is the worst, which is 9.56 km x L(-1). The driving cycles used on abroad can not reflect the actual driving conditions in China. PMID- 19558129 TI - [Influence of traffic restriction on road and construction fugitive dust]. AB - By monitoring the road and construction dust fall continuously during the "Good Luck Beijing" sport events, the reduction of road and construction dust fall caused by traffic restriction was studied. The contribution rate of road and construction dust to particulate matter of Beijing atmosphere environment, and the emission ratio of it to total local PM10 emission were analyzed. The results show that the traffic restriction reduces road and construction dust fall significantly. The dust fall average value of ring roads was 0.27 g x (m2 x d)( 1) in the "traffic restriction" period, and the values were 0.81 and 0.59 g x (m2 x d)(-1) 1 month and 7 days before. The dust fall average value of major arterial and minor arterial was 0.21 g x (m2 x d)(-1) in the "traffic restriction" period, and the values were 0.54 and 0.58 g x (m2 x d)(-1) 1 month and 7 days before. The roads emission reduced 60%-70% compared with before traffic restriction. The dust fall average values of civil architecture and utility architecture were 0.61 and 1.06 g x (m2 x d)(-1) in the "traffic restriction" period, and the values were 1.15 and 1.55 g x (m2 x d)(-1) 20 days before. The construction dust reduced 30% 47% compared with 20 days before traffic restriction. Road and construction dust emission are the main source of atmosphere particulate matter in Beijing, and its contribution to ambient PM10 concentration is 21%-36%. PM10 emitted from roads and constructions account for 42%-72% and 30%-51% of local emission while the local PM10 account for 50% and 70% of the total emission. PMID- 19558130 TI - [Different leaching procedures for heavy metal toxicity of waste PCBs]. AB - The purpose of this paper is to evaluate the hazardous of heavy metals of waste printed circuit boards (PCBs) by using four kinds of methods (GB 5086.1-1997, GB 5086.2-1997 from China and 1311 (TCLP), 1312 (SPLP) from USA), and the four kinds of methods are compared to choose a better way to identify toxicity characteristics of waste PCBs. The results show that the leaching concentrations of heavy metals (Cu, Pb, Ni, Mn, Zn and Fe) in the leachate after TCLP are much higher than that after the other three kinds of methods, the concentration of Pb is over the standards, so the waste PCBs may be the hazardous solid waste, the TCLP could be the appropriate way to assess the heavy metals hazardous characteristics of waste PCBs. The leaching concentrations by the leaching methods used in China are relatively lower, which could be disadvantageous to control the hazardous wastes. The initial pH and particle size of waste PCBs have great effect on the leachability of heavy metals from waste PCBs. PMID- 19558131 TI - [Release amount of heavy metals in cement product from co-processing waste in cement kiln]. AB - Clinker was produced by Simulating cement calcination test, and concrete samples were also prepared according to national standard GB/T 17671-1999. Long-term cumulative release amount of heavy metals in cement product from co-processing waste in cement kiln was researched through leaching test which refers to EA NEN 7371 and EA NEN 7375, and one-dimensional diffusion model which is on the base of Fick diffusion law. The results show that availabilities of heavy metals are lower than the total amounts in concrete. The diffusion coefficients of heavy metals are different (Cr > As > Ni > Cd). During 30 years service, the cumulative release amounts of Cr, As, Ni and Cd are 4.43 mg/kg, 0.46 mg/kg, 1.50 mg/kg and 0.02 mg/kg, respectively, and the ratios of release which is the division of cumulative release amount and availability are 27.0%, 18.0%, 3.0% and 0.2%, respectively. The most important influence factor of cumulative release amount of heavy metal is the diffusion coefficient, and it is correlative to cumulative release amount. The diffusion coefficient of Cr and As should be controlled exactly in the processing of input the cement-kiln. PMID- 19558132 TI - [PCDD/F emission characteristics in different waste incineration facilities and evaluation of 23478-PeCDF as I-TEQ indicator of PCDD/F in flue gases]. AB - The emission characteristics of PCDD/F from different waste incineration facilities in China, the correlations of different PCDD/F isomers with I-TEQ and the linear regression analysis of 23478-PeCDF with I-TEQ were reported in this article. Statistical analysis of the homologue patterns of PCDD/F in different incineration facilities will provide valuable information in environmental survey to find the contamination sources. PCDD/F congener patterns were almost constant among municipal and hazardous waste incineration facilities with only very small variations, but those patterns in medical waste incineration facilities were slightly different from medical and hazardous waste incineration facilities. It was found that the major contributors to the toxic PCDD/F concentrations were OCDD and 1234678-HpCDF contributing for 12.3%-23.0% and 15.0%-19.7% respectively. However, the largest contributor to I-TEQ was 23478-PeCDF and 33.1%-34.5% of I TEQ was the presence of this congener. Correlation analysis showed that:23478 PeCDF posed the best the correlation relationship with I-TEQ, R2 0.93-0.99. The most toxic 2378-TCDD only posed weak correlation with I-TEQ, R2 0.29-0.49. The correlation coefficient between the most abundant OCDD and I-TEQ was only 0.03 0.12. However, another abundant congener 1234678-HpCDF posed better correlation with I-TEQ, R2 0.62-0.87. Linear regression analysis showed that even I-TEQ fell into the concentration range of 5-6 orders of magnitudes, the linear relationship between 23478-PeCDF and I-TEQ was still excellent. The slopes of that lines, in different waste incineration facilities, were in the range of 1.16-1.40 and R2 0.94-0.97. PMID- 19558133 TI - [Hydrothermal pretreatment of canteen waste for grease removal and digestibility improvement]. AB - Canteen waste was hydrothermal pretreated to investigate the characters of degreasing, hydrolysis and digestibility by biochemical methane assays. Liquidization and hydrolysis were enhanced in hydrothermal pretreatment. pH of hydrolysate decreased, and SCOD and VFA increased significantly followed by increasing hydrothermal temperature. Floatable grease increased to appropriate ratio for removal, and digestion process completed in 4 days to 6 days with double degradation rate after pretreatment at 150 degrees C for 60 minutes. The 4 day incubation biogas yield was maximum after pretreatment at 170 degrees C, and the methane potential, however, was decreased at higher pretreatment temperature. Energy consumption of optimized pretreatment was recoverable with energy balance analysis. PMID- 19558134 TI - [Degradation of cyanide and maturity in cassava processing wastes composting]. AB - An investigation was carried out to approach the degradation of cyanide and maturity during the cassava processing wastes composting process. Mixtures of cassava hull, cassava residues and pig manure were used in the experiment. Parameters like temperature, pH, cyanide, cellulose, hemicellulose, lignin and C/N ratio were assessed during the composting process, the effect of composting process on the degradation of cyanide and maturity were evaluated. The results reveal that the content of cyanide decreases sharply and declines to 2.08 mg/kg (30 days of composting), the degradation rate of cyanide is 94.16% and is in accord with food safety standard. After 15 days of the composting process, degradation of composting materials containing carbon (starch, cellulose, hemicellulose) and cyanide are quick and the degradation rates of them are more than 80%, properties tend towards stability basically. During 30 days of the composting process, the composting temperature drops to normal temperature and tends to stability, pH remains stable at 7.2. Parameters like C/N ratio, nitrate nitrogen (NO3(-)-N) and ammonia nitrogen (NH4(+)-N) as maturity evaluation index were measured, and the results indicate that physical and chemical properties keep stability after 15 days of cassava processing wastes composting process. At the end of fermentation, C/N ratio is 17.55, the content of nitrate-nitrogen and ammonia nitrogen reach 2.5g/kg and 10 mg/kg respectively, NO3(-)-N/NH4(+)-N ratio is 250. The changes of these above mentioned parameters meet with maturity evaluation standard. Proving that cassava processing wastes during 30 days of composting treatment can achieve stability and security state. PMID- 19558135 TI - [Pharmacology: my whole life career]. PMID- 19558136 TI - [Therapeutic cloning and somatic cell reprogramming: every road leads to Rome]. AB - The researches of therapeutic cloning and somatic cell reprogramming, two strategies used to generate patient-specific autologous stem cells, have recently made great progress. Therapeutic cloning refers to derivation of embryonic stem cells from blastocyst produced by somatic cell nuclear transfer, whereas somatic cell reprogramming refers to establishment of induced pluripotent stem (iPS) cells from differentiated somatic cells by ectopic expression of specific transcription factors. The two strategies differ in their methodological approaches, technical obstacles and ethical debates, but confront similar problems including the differentiation of stem cells and the feasibility of cell replacement therapy. This review discusses the research advance of these two biotechnologies and summarizes their difference and similarity. PMID- 19558137 TI - [Endoplasmic reticulum stress and heart diseases]. AB - Endoplasmic reticulum (ER) is a multifunctional organelle responsible for the synthesis and folding of proteins, calcium storage and synthesis of lipid. Perturbations of ER functional homeostasis result in accumulation of misfolded or unfolded protein in ER, which induces a series of cellular responses (ER stress) through proportional signaling pathway. It is reported that ER stress contributes to development of diseases of heart, including myocardial ischemia-reperfusion injury, cardiomyopathy and heart failure, et al. ER stress might be a novel therapeutic target in heart diseases. PMID- 19558138 TI - [Research progress of cardiac systems biology]. AB - Systems Biology is one of the most widely discussed fields among emerging post genomic disciplines. Medical systems biology is an important component of systems biology. The goals of medical systems biology are gaining a complete understanding of human body in normal and disease states. Driven by the great importance of cardiovascular diseases, cardiac systems biology is improving rapidly. This review provides an overview of major themes in the developing field of cardiac systems biology, including some of the high-throughput experiments and strategies used to integrate the datasets, various types of computational approaches used for developing useful quantitative models, and successful examples, future directions of cardiac systems biology. PMID- 19558139 TI - [Development of nano-scale genic carriers in biomedicine]. AB - Nano biotechnology is a developing field of science and technology in the 21st century. The emergence of nanomaterials provides a new way for gene carriers. This paper demonstrates the advantages and types of nano-scale genic carriers and the methods for introducing it into the body, and reviewed the application of nano-scale genic carriers in gene therapy and in breeding. PMID- 19558140 TI - [The interactions between the circadian clock and aging]. AB - It is well known that almost all organisms ranging from single cell creatures to human beings exhibit circadian rhythms in physiology and behavior under the control of the internal circadian clock. The internal circadian clock is composed of a master clock which is localized in the suprachiasmatic nucleus and the peripheral clocks located in peripheral tissues such as the liver and heart. Along with aging, the circadian rhythm alters in many aspects, including the amplitude, free-running period and the expression phase. On the other hand, the circadian clock also influences the process of aging. The disorganized circadian rhythm accelerates the aging process. This article briefly reviews the recent progress in the interactions between the circadian clock and aging, and provides evidence to further understand the mechanism of aging and the impact of aging on the organisms. PMID- 19558141 TI - [Physiological and molecular control of lipid accumulation in Caenorhabditis elegans]. AB - Fat storage is a complex physiological process, and model organism of Caenorhabditis elegans has already been explored as an important model to study lipid accumulation. The lipid particles or fatty acid can be stained or labeled with Sudan Black B or Neil Red. The metabolism pathways for fatty acid synthesis and breakdown in nematodes are almost identical to those in other organisms, and functions of many genes encoding the key regulation enzymes have been identified. At least four central regulation pathways are involved in the fat storage control in nematodes: insulin and TGF-beta signaling pathway, sbp-1/mdt-15 mediated pathway, nhr-49 mediated pathway, and TOR and hexosamine pathway. Moreover, neurotransmitters 5-HT, dopamine, and glutamate were found to participate in the control of lipid accumulation. In addition, involvement of tub-1 and bbs-1 in neuronal control of fat storage suggest the possibly important roles of amphid structure and sensory neurons in regulating lipid accumulation. The data obtained in C. elegans on fat storage control will contribute largely to the study on metabolism related diseases, such as obesity, in human beings. PMID- 19558142 TI - [Glucose regulated protein 78 kD]. AB - One of the most important chaperones located on endoplasmic reticulum, GRP78, referred as BiP (immunoglobulin heavy chain binding protein), belongs to heat shock protein 70 family. GRP78 exists conservatively among a wide variety of biological species, and acts as a central regulator of endoplasmic reticulum (ER) functions, participating in ER protein folding and assembly process, and maintaining ER Ca2+ homeostasis, unfolded protein response and specific anti apoptotic actions. Specific regulatory cis-elements such as ER stress response element (ERSE) and cAMP response element (CRE) were identified on the promoter of GRP78. Dynamic epigenetic interactions between specific transcription factors such as AFT6 and regulatory elements in GRP78 gene promoter might contribute to human GRP78 constitutive or inducible transcription, resulting from some physiological and pathological stresses. Recently, cellular relationship between GRP78 expression and hepatic steatosis, cancer and nervous system diseases in human was underwent further clinical and biochemical research, which will benefit to human beings. PMID- 19558143 TI - [Dedifferentiation of mammalian neural cells and its induction]. AB - Dedifferentiation of cells has elicited a great deal of attention in the field of cell biology. A great quantity of studies have proved that many types of cells, such as neuron, Schwann cell, astrocyte, oligodendrocyte, etc., within mammalian nervous system could dedifferentiate in different extent, featured in cell becoming immature, reentering cell cycle and regaining pluripotential. This review concerns on dedifferentiation of various types of cells within mammalian nervous system and inducing conditions, as well as its effect on neural regeneration. PMID- 19558144 TI - [Recent advances on liver X receptors]. AB - The liver X receptors (LXRs), that are nuclear receptors activated by oxysterols, play central roles in the transcriptional control of cholesterol metabolism and function as the sensors of cholesterol homeostasis. In addition, LXRs have also been found to modulate lipogenesis, carbohydrate metabolism, innate immune and inflammatory responses in macrophages. Hence, the ability of LXRs to integrate metabolic and inflammatory signaling makes them particularly attractive targets for intervention in human atherosclerosis, hypercholesterolemia and type 2 diabetes. PMID- 19558145 TI - [New advancement of genetically manipulated mouse models in dyslipidemia and atherosclerosis]. PMID- 19558146 TI - [The role of SUMO in regulation of NF-kappaB signaling pathway]. PMID- 19558147 TI - [Association of resistin with obesity with type 2 diabetes]. PMID- 19558148 TI - [The molecular mechanism of Ca2+ -dependent facilitation and Ca2+ -dependent inactivation in voltage-gated Ca2+ channels]. PMID- 19558149 TI - [Research progress about calreticulin in myocardial hypertrophy]. PMID- 19558150 TI - [Progress of the relationship between fibrin (ogen) and atherosclerosis]. PMID- 19558151 TI - [Recent advances on the roles of cPLA2 in CNS impairment]. PMID- 19558152 TI - [Regulation of heat shock factor 1 activity]. PMID- 19558153 TI - [Progesterone and the pain perception modulation]. PMID- 19558154 TI - [The progress on transmembrane protease serines]. PMID- 19558155 TI - [The signaling transduction of Smads during the myocardiocyte apoptosis]. PMID- 19558156 TI - [Trilobate technique, a new principal to repair cleft lip]. AB - OBJECTIVE: To develop a new method for reparation of cleft lip, and to evoke more colleagues for advance practices and study, in order to determine her indication and contraindication as soon as possible. METHODS: 48 cases were included into this study. Trilobate flap were designed in floor of nose and lip area in cleft side, rotate two of the three flaps upwards, respectively to elevate the tip of nose, and to reconstruct the floor of nose. As for the left flap, it was derived transversally to opposing side, sutured with the flap of non-cleft-side. RESULTS: With this technique, less tissue was lost, better vertical lengthening and good formed cuspids-bow was achieved, and the scar was a parallel line being symmetry to the philtrum column opposite. Meanwhile, because the tension was mainly located in the area where there was no mini flaps, the blood supply was good enough, rarely occur any necrosis in the tip of flaps. All cases in this study obtained perfect healing, with good appearance at nostrils and floor of nose. CONCLUSIONS: In use of the method of trilobate flap, we can draw down the peak of the cuspids bow effectually, hence avoid the addition cut in the lower part of the lip, decrease the scar on skin, as well as nice reconstruction of floor of nose, philtrum column and nostril. Because lack of long term study, we evoke more colleagues for cooperation in advance study. PMID- 19558157 TI - [Anatomic and functional plasty for unilateral complete cleft lip]. AB - OBJECTIVE: To explore methods for one-stage repairing of nose deformity and alveolar cleft, as well as primary unilateral complete cleft lip. METHODS: The nose deformity and alveolar cleft were repaired by Carstens' surgical methods, rhBMP-2 was used for correction of alveolar cleft. RESULTS: From 2004 to 2008, 45 cases of cleft lip were treated in the United States and followed up for 3-5 years. In 2007, 10 cases of cleft lip were treated in China and followed up for 1 year. All results were satisfactory. CONCLUSIONS: Carstens' surgical methods can be applied for early correction of nose deformity and alveolar cleft in patients with complete cleft lip. PMID- 19558158 TI - [The nasal development after one-staged correction of nose deformity and unilateral complete cleft lip in infancy]. AB - OBJECTIVE: To introduce one-staged correction of nasal deformity and unilateral complete cleft lip in infancy and to observe the nasal development after the operation. METHODS: The unilateral complete cleft lip and nasal deformity were corrected in one stage in27 cases. They were followed up for several years. With post-operative photos, the anthropometric method was used to analyze the nasal development. RESULTS: The long-term results were excellent in 10 cases, good in 14 cases, and poor in 3 cases. CONCLUSIONS: Based on the anatomic findings of nasal blood supply, one-staged correction of nasal deformity and unilateral complete cleft lip in infancy can be performed with no obvious interference with nasal development. The secondary nasal deformity before school age can be alleviated or avoided. PMID- 19558159 TI - [One-staged repair of unilateral cleft lip and nasal deformity with modified Mohler]. AB - OBJECTIVE: To study the therapeutic effect of one-staged repair of unilateral cleft lip an nasal deformity with modified Mohler. METHODS: The unilateral cleft lip and nasal deformity were repaired in one stage with modified Mohler. The nasal retainers were kept for at least 6 months. The patients were followed up at 3 months and 12 months after operation. RESULTS: From Sept. 1998 to Aug. 2008, 103 cases, aged from 2.5 m to 12 m, were treated. 3 months after operation, 97 of 103 patients had good appearance of both lips and noses, which included unobvious scars, symmetric Cupid's bows and nostrils, intermedial nasal columellae and eminentia nasal tips. 6 of them had good appearance of noses only with a little malposition of Cupid' s bows. 89 patients were followed up for 12 months with no deterioration of nasal deformity and delayed growth of greater alar cartilages. 3 patients who had malposition of Cupid's bows also got better. CONCLUSIONS: The one-staged repair of unilateral cleft lip and nasal deformity with modified Mohler is very practical. PMID- 19558160 TI - [Application of tissue granule of oral mucous in plastic surgery]. AB - OBJECTIVE: To study the application of tissue granule of oral mucous in plastic surgery. METHODS: Tissue granule of oral mucous was placed on glutin sponge and they were used for urethral (12 cases) and vaginal (14 cases) reconstruction in 26 cases. RESULTS: Satisfactory results were achieved in 24 cases. One case of urethral fistula and one case of urethral meatus stricture were happened. The two cases underwent secondary operation. CONCLUSIONS: Tissue granule of oral mucous membrane is good supply for repairing mucous defect. It has the advantages of high growth and survival rate, and less shrinkage. It is useful for urethral or vaginal reconstruction which are covered with mucous membrane. PMID- 19558161 TI - [Staged treatment of the severe hypospadias in adult]. AB - OBJECTIVE: To report the treatment of serious hypospadias in adults with free graft of tubed mouth mucosa and scrotal fascia flaps. METHODS: The tubed mouth mucosa was free grafted to fabricate the distal segment of urethra. It was anastomosed to the urethra at the second stage. The scrotal fascia flap was used to cover the penile wound. The biggest flap was 3 cm in width and 6.5 cm in length. RESULTS: From Jan. 2002 to Dec. 2007, 76 adults with severe hypospadias were treated. Infection happened in 4 cases. 2 cases had urethral fistula due to the partial flap necrosis which was healed automatically within 2-4 weeks. All the other patients healed primarily. CONCLUSIONS: It is a good method for the treatment of serious hypospadias in adults with scrotal fascia flaps and free graft of tubed mouth mucosa which is anastomosed to the urethra at the second stage. PMID- 19558162 TI - [Distraction osteogenesis of the mandibular ramus in hemifacial microsomia]. AB - OBJECTIVE: To study the application of distraction osteogenesis (DO) of the mandibular ramus in hemifacial microsomia. METHODS: From 1999 to 2006, 15 patients with hemifacial microsomia were treated with DO of the ramus, including 12 cases of children and juveniles, 3 cases of grown-ups. Maxillary Le Fort I osteotomy, unilateral sagittal split ramus osteotomy, genioplasty and free scapular flaps were selected to perform on the first or secondary stage of treatment. RESULTS: Successful distraction was achieved in all patients with an average distraction distance of 21.25 mm. The distractor was removed after consolidation proved by clinical examination and X-ray. Facial appearance and and occlusal relationship were improved greatly. The occlusal plane was almost normal. There was no infection, malunion or permanent injury of inferior alveolar nerve. CONCLUSIONS: DO of the ramus is very effective for the correction of severe hemifacial microsomia. It has much better therapeutic effect than traditional orthognathic procedures. PMID- 19558163 TI - [Immediate reconstruction of tissue defects on eyelid caused by tumor excision]. AB - OBJECTIVE: To introduce the methods for immediate reconstruction of tissue defects on eyelid caused by tumor excision. METHODS: Single or combined local skin flaps were used to repair tissue defects on eyelids after tumor excision. RESULTS: Since 1994, 216 patients (224 sides) were treated. All flaps survived with primary healing. 149 patients were followed up for 3 months to 10 years with satisfactory results. 2 patients with squamous cell carcinoma recurred and reoperated with local skin flap again. CONCLUSIONS: It is a simple, safe, flexible and reliable method to immediately repair the tissue defects with local flaps on eyelid caused by tumor excision. PMID- 19558164 TI - [Correction of severe depression deformities at lower eyelid]. AB - OBJECTIVE: To evaluate the application of temporal fascia flaps in the correction of severe depression deformities at lower eyelids. METHODS: Severe depression deformities at lower eyelid were corrected with temporal fascia flaps pedicled with superficial temporal artery in 9 cases. RESULTS: All flaps survived with good appearance. All patients were followed up for 6-12 months with good long term results. The donor sites had no obvious scalp scar. CONCLUSIONS: Temporal fascia flap is an optimal choice for correction of the severe depression deformities at lower eyelid. It is easily performed with good result and less donor site morbidity. PMID- 19558166 TI - [Three-dimensional finite element analysis for different directions distraction at midface]. AB - OBJECTIVE: To investigate the biomechanical changes with different directions distraction at midface. METHODS: An anteriorly directed 500 g force was applied to the floor of apertura piriforms in different directions to the occlusal plane. Three-dimensional finite element analysis was used to evaluate the biomechanical change of craniofacial complex. RESULTS: As the force direction was moved downward, the sagittal distraction length of the craniofacial complex decreased and vertical movement changed from upward to downward. The craniofacial complex was moved anteriorly when the downward force was applied about 20-30 degrees to the occlusal plane. The forces could generate the uniform stress distribution in the craniofacial sutures and avoid counterclockwise rotation of the maxilla. CONCLUSIONS: The craniofacial complex can be effectively distracted anteriorly when the downward force is applied to the floor of aperture piriforms in direction of 20-30 degrees to the occlusal plane. PMID- 19558165 TI - [Thoracoscopy assisted Nuss procedure for pectus excavatum correction]. AB - OBJECTIVE: To evaluate the short-term effect and experience of Nuss procedure on 120 cases of patients with pectus excavatum. METHODS: Thoracoscopy assisted Nuss procedure with different ways of anesthesia were applied to 120 cases of patients with pectus excavatum, including 7 cases of recurrence after traditional surgical procedure (6 cases) and Nuss method (another one). The patients ranged in age from 2.5 to 43 (mean 14.1) years and in Haller index from 2.91 to 29. Of the 120, 73 had symmetric and 47 had asymmetric pectus excavatum. The Nuss procedure is performed with general anesthesia and a convex steel bar is inserted under the sternum with thoracoscopy through small bilateral thoracic incisions. The steel bar is inserted with the convexity facing posteriorly, and when it is in position, the bar is turned over, thereby correcting the deformity. RESULTS: The operation was successfully accomplished without severe complications in all the 120 cases. The mean operative time was 58 minutes and the mean volume of blood loss was 30 ml. 103 patients had one bar inserted while the other 17 cases with more extremely diffuse depression required 2 or even 3 bars to get a satisfactory correction. Such methods as modifications to the fixing points and the shape of the bar, partial osteotomy, were developed to deal with asymmetric ones. CONCLUSION: The Nuss procedure is a minimally invasive technique for correction of pectus excavatum. It can lead to a satisfactory outcome and surgical time is less. PMID- 19558167 TI - [Experimental study of the effect of liposome-mediated vascular endothelial growth factor gene on the flap survival in rats]. AB - OBJECTIVE: To investigate the effect of VEGF gene on the random flap survival after pedicle division at different time in rats. METHODS: The random-pattern flaps were formed on the back of the 120 SD rats. PcDNAVEGF165 (gene) wrapped with liposome was injected into the flaps in experimental group (n = 40). The flaps in the two control groups were injected with PcDNA (n = 40) or saline (n = 40). 1, 3, 5, 7 days after injection, 10 rats in each group were randomly selected to performed pedicle division. 7 days after pedicle division, the rats were killed to measure the flap survival rate. The microvessels was studied by histologic examination. The expression of VEGF was assessed by immunohistochemical staining. The flaps were also examined under the electron ultrastructure microscopy. RESULTS: 1) Flap survival rate after pedicle division in experimental group at 1 day, 3 days, 5 days, 7 days after injection, were (45.45 +/- 12.24)%, (82.95 +/- 3.81)%, (85.00 +/- 3.38)%, (85.96 +/- 3.25)%, respectively. The flap survival rates were significant different between experimental group and the control groups at 3, 5, 7 days after injection (P < 0.05), but not at 1 days after injection (P > 0.05). 2) The average microvascular diameter and number in experimental group were significantly higher than those in control groups (P < 0.05). 3) The expression of VEGF in experimental group was significantly higher than that in the two control groups (P < 0.05). 4) Ultrastructure study showed more angiogenesis in experimental group. CONCLUSIONS: Subcutaneous injection of liposome-mediated VEGF gene can increase the survival rate of flap with early pedicle division. It is a simple, efficient, economic, and the relatively safe gene therapy. PMID- 19558168 TI - [Constructing tissue engineered trachea-like cartilage graft in vitro by using bone marrow stromal cells sheet and PLGA internal support: experimental study in bioreactor]. AB - OBJECTIVE: To explore the feasibility of constructing tissue engineered trachea like cartilage graft in vitro by using bone marrow stromal cells (BMSCs) sheet and PLGA internal support. METHODS: Rabbit BMSCs were expanded and induced by transforming growth factor-1 to improve chondrocyte phenotype of BMSCs. BMSCs sheets were obtained by continuous culture and wrapped the PGLA scaffold in the shape of cylinder. The constructs were incubated in spinner flask for 8 weeks and cartilage formation was investigated by gross inspection, histology, glycosaminoglycan and mechanical strength content. RESULTS: After in vitro culture, cartilage like tissue in cylindrical shape had been regenerated successfully. Stiff, shiny, pearly opalescence tissues were observed. Histological analysis showed engineered trachea cartilage consisted of evenly spaced lacunae embedded in matrix, cells stationed in the lacunae could be noticed clearly. Safranin-O staining on the sections showed homogenous and positive red staining, which demonstrated that the engineered tissue was rich in proteoglycans. CONCLUSIONS: Based on the cell sheet and internal support strategy, trachea-like cartilage in cylindrical shape could be successfully fabricated which provided a highly effective cartilage graft substitute and could be useful in many situations of trachea-cartilage loss encountered in clinical practice. PMID- 19558169 TI - [Experimental study of the effect of adipose tissue derived stem cells on the survival rate of free fat transplantation]. AB - OBJECTIVE: To investigate the feasibility of using adipose tissue derived stem cells (ASCs) to promote neovascularization and survival rate of free fat transplantation. METHODS: ASCs were isolated from aspirates from human liposuction and cultured in vitro. The cells were incubated in adipogenic, osteogenic, and chondrogenic medium for 2-4 weeks to induce adipogenesis, osteogenesis and chondrogenesis, respectively. ASCs were labelled by DiI. ASCs (A group), Insulin (B group), Medium (C group) were respectively mixed with free fat graft from aspirates. The mixtures were injected subcutaneously at the three random points on the back of eighteen 4- 6-week-old nude mice. Transplanted fat tissue was harvested after 6 months. The grafts were assessed by morphological observation, HE staining and immunohistochemistry. RESULTS: ASCs can be easily harvested from liposuction aspirates and differentiate into adipogenic, osteogenic, chondrogenic lineages. The wet weight of transplanted fat tissue in ASCs group was (165.97 +/- 5.51) mg, significantly higher than that in the insulin group (93.42 +/- 5.12) mg and control group (67.64 +/- 5.09) mg (P = 0.000). The rate of fibrosis and steatonecrosis in ASCs group was( 152.2 +/- 9.8)/10HF, significantly lower than that in the Insulin group (743.9 +/- 20.4)/10HF and control group (892.2 +/- 16.5)/10HF (P = 0.000). DiI labelled ASCs were found between adipocytes and in the connective tissue in free transplanted fat tissue, and some of these cells were immunopositive for antihuman CD31 and FITC, suggesting differentiation into vascular endothelial cells. CONCLUSIONS: ASCs can differentiate into vascular endothelial cells and contribute to angiogenesis in free transplanted fat tissue. ASCs can increase the survival rate and decrease the rate of fibrosis and steatonecrosis of free transplanted fat tissue. These findings suggest that ASCs-assisted transplantation may be an ideal cell therapy. PMID- 19558170 TI - [Expression of hypoxia-inducible factor-1 alpha and vascular endothelial growth factor in malignant melanoma]. AB - OBJECTIVE: To evaluate the expression of hypoxia-inducible factor-1 alpha (HIF-1 alpha) and vascular endothelial growth factor (VEGF) and their relationship in malignant melanoma. METHODS: 32 cases of malignant melanoma and 11 samples of normal skin were examined for HIF-1 alpha and VEGF expression by immunohistochemistry. RESULTS: The expression rate of HIF-1 alpha and VEGF was 62.5% (20/32) and 84.8% (27/32), respectively, in malignant melanoma, and 9.1% (1/11) and 18.2% (2/11), respectively, in normal skin, showing a significant difference between the two groups. Positive relationship between the expression of HIF-1 alpha and VEGF was found in malignant melanoma (P < 0.05). CONCLUSIONS: 1) The high expression of HIF-1 alpha and VEGF in malignant melanoma indicates that it may play an important role in the generation and development of malignant melanoma. 2) The positive correlation between the expression of HIF-1 alpha and VEGF in malignant melanoma suggests that HIF-1 alpha may be involved in the regulation of VEGF in malignant melanoma. PMID- 19558172 TI - [Experimental study of PVPP/silicone composite automatic expanded material as implants]. AB - OBJECTIVE: To study the feasibility of Polyvinylpolypyrrolidone (PVPP)/silicone composite automatic expanded material as implants. METHODS: The PVPP hydrogel was mixed with silicone through the location at the high temperature. Implants with different ratio of PVPP to silicone were placed under the back and nose skin in 24 New Zealand rabbits. The surrounding tissue reaction, material and skin expansion were observed and compared with those with pure silicone implants. RESULTS: The study lasted for 200 days. Compared with pure silicone implants, the composite material could expand automatically and stop expanding at about 2 weeks after implantation. Histological study showed similar inflectional and foreign body reaction around the composite material and the pure silicone. CONCLUSIONS: Compared with pure silicone, the PVPP/silicone composite implant has the advantage of automatic expansion, so as to expand the soft tissue. PMID- 19558174 TI - Determination of rapid-equilibrium kinetic parameters of ordered and random enzyme-catalyzed reaction A+B=P+Q. AB - This article deals with the rapid-equilibrium kinetics of the forward and reverse reactions together for the ordered and random enzyme-catalyzed A+B=P+Q and emphasizes the importance of reporting the values of the full set of equilibrium constants. Equilibrium constants that are not in the rate equation can be calculated for random mechanisms using thermodynamic cycles. This treatment is based on the use of a computer to derive rate equations for three mechanisms and to estimate the kinetic parameters with the minimum number of velocity measurements. The most general of these three programs is the one to use first when the mechanism for A+B=P+Q is studied for the first time. This article shows the effects of experimental errors in velocity measurements on the values of the kinetic parameters and on the apparent equilibrium constant calculated using the Haldane relation. PMID- 19558176 TI - Determination of the g tensors for the dominant stable radicals in X-irradiated beta-D-fructose single crystals. AB - In spite of recent successful identifications of radicals produced after X-ray irradiation at 10 and 77 K in beta-D-fructose, the structure of the two stable radicals dominating the electron paramagnetic resonance (EPR) spectrum after room temperature irradiation is still unclear. Based on the agreement between proton hyperfine (HF) tensors obtained in electron nuclear double resonance (ENDOR) experiments and the results of single molecule density functional calculations, a model for these radicals, involving OH abstraction at the C2 ring position, had previously been proposed, but this assignment could not be confirmed when the radical was embedded in a crystal environment. In this paper, we therefore provide additional experimental information for these radicals. First, their g tensors are determined from angular dependent ENDOR-induced EPR experiments. The relatively large anisotropy of these tensors is indicative of delocalization of the unpaired electron onto a neighboring oxygen atom. Second, EPR spectra of fructose powders, selectively enriched in (13)C on various ring positions, are presented, demonstrating that the HF interaction with the carbon atom C3 is larger than with the C2. Combining the g tensor, proton and (13)C HF data, we conclude that the structure of the stable radicals differs strongly from that of intact molecules and that further advanced quantum chemical modeling will be required to fully identify them. PMID- 19558175 TI - Evidence of high *OH radical quenching efficiency by vitamin B6. AB - Molecules acting as antioxidants capable of scavenging reactive oxygen species (ROS) are of the utmost importance in the living cell. The antioxidative properties of pyridoxine (vitamin B6) have recently been discovered. Previous theoretical calculations have shown a high reactivity of pyridoxine toward hydroxyl radicals, where the latter preferably abstract H from either carbon of the two methanol substituents (C8 or C9). In this study, we have explored the reactivity of pyridoxine toward further hydroxyl radicals, considering as the first step the H abstraction from either C8 or C9, also including addition reactions and cyclization. Many of the reactions display similar DeltaG, and hence, the quenching of hydroxyl radicals by pyridoxine may undergo different pathways leading to a mix of products. In addition, we observe that pyridoxine, under high hydroxyl radical concentrations, may scavenge up to eight radicals, supporting its observed high antioxidant activity. PMID- 19558171 TI - [The expression of aromatase P450 in the mammary gland of hypertrophic breast]. AB - OBJECTIVE: To investigate the expression of aromatase P450 in the mammary gland of hypertrophic breast. METHODS: The expression of aromatase P450 in the mammary gland was detected by SP immunohistochemistry in 28 cases of hypertrophic breasts and 12 cases of normal volume breasts. The results were analysed by SPSS 13.0, and comparison among groups was performed by Fisher's exact test. RESULTS: The positive expression rate of aromatase P450 in hypertrophic group was 39.29%, while there was no positive expression in normal group, showing a significant difference between the two groups (P < 0.05). There was no significant difference between the gland hyperplasia group and adipose hyperplasia group (P > 0.05). There were also no significant difference between the groups with different lactation history or different extent of hypertrophy and mastocytosis (P > 0.05). CONCLUSIONS: Overexpression of aromatase P450 in mammary gland may be related to the development of hypertrophic breast. PMID- 19558177 TI - Organo-selenium induced radical ring-opening intramolecular cyclization or electrophilic cyclization of 2- (arylmethylene)cyclopropylaldehyde: a tunable synthesis of 1-naphthaldehydes or 3-oxabicyclo[3.1.0]hexan-2-ols. AB - 1-Naphthaldehydes and 3-oxabicyclo[3.1.0]hexan-2-ols can be prepared, respectively, by the intramolecular alkylation and cyclization of (E)-2 (arylmethylene)cyclopropylaldehyde 1 mediated by different organo-selenium reagents. The properties of selenium reagents may play an important role in the reactions. A rationale for these transformations is proposed. PMID- 19558178 TI - A DFT study on the mechanism of the annulation reaction of trichloronitroethylene with aniline in the synthesis of quinoxalinone-N-oxides. AB - The new annulation reaction of trichloronitroethylene with aniline results in the formation of a quinoxalinone-N-oxide derivative. The mechanism of this one-pot annulation reaction between trichloronitroethylene (TCNiE) and anilines has been extensively investigated with B3LYP/6-31+G** methodology. Five different paths (1 5) were proposed and modeled by using this method. These paths were compared in terms of the activation energies of their rate-determining steps and in regard to the experimental findings. Paths 3 and 5, proceeding via four-membered heterocyclic rings, were found to be the most plausible paths with activation energies of 32 and 29 kcal/mol for the rate-determining steps, respectively. The effects of substituent, solvent, temperature, and computational method on these steps were also investigated. The results showed that path 5 is the most plausible mechanism for the annulation reaction of trichloronitroethylene with aniline. PMID- 19558179 TI - Dynamic (1)H NMR spectroscopic study of the ring inversion in N-sulfonyl morpholines--studies on N-S interactions. AB - The effect of the exocyclic conjugation, via d-p orbital interaction and/or negative hyperconjugation (anomeric effect) of the N-S bond, on the inversion of the morpholine ring in some N-arylsulfonyl morpholines is studied by variable temperature (1)H NMR spectroscopy in different solvents. The observed free energy barriers are 9.2-10.3 kcal mol(-1); the lower values were obtained with increasing conjugation (substituents of higher electron withdrawing power) along the series. The barrier to ring inversion of 1e was solvent independent. X-ray data of compounds 1b,d reveal the chair conformation of the six-membered ring, the flattened pyramidal orientation of the ring nitrogen atom, and the sulfonyl group in equatorial position with the plane containing the C(aryl)-S-N bond perpendicular to the plane of the benzene ring. In addition, the sulfonamide group prefers a conformation with the S-C bond antiperiplanar with respect to the nitrogen atom lone pair and the -CH(2)-N-CH(2)- moieties in staggered conformation with the S-O bonds of the SO(2) group. PMID- 19558180 TI - Evidence for effective p(Z)-pi(Ar) conjugations (Z = S, Se, and Te, as well as Z = O) in 9-(arylchalcogenyl)triptycenes: experimental and theoretical investigations. AB - The p(Z)-pi(Ar) conjugations must operate fully in the ground states of 9 (arylchalcogenyl)triptycenes (p-YC(6)H(4)ZTpc:1 (Z = O), 2 (Z = S), 3 (Z = Se), and 4 (Z = Te)), where the p-YC(6)H(4) group is placed in the bisected area between two phenyl planes of the triptycyl group with the parallel orientation. The ground-state geometries, which we call (A: pl), are confirmed by X-ray analysis. However, the conjugations never operate in the transition states between (A: pl) and/or the topomeric structures (A': pl'), where the Z-C(Tpc) bond is perpendicular to the plane. The site-exchange processes correlate to the conjugations. Temperature-dependent (1)H NMR spectra are analyzed for 2 and 3 to demonstrate the effective p(Z)-pi(Ar) conjugations. The activation energies for the interconversion between (A: pl) and (A': pl') (GR: gear process) were obtained for 2 (DeltaG(GR)(2)) and 3 (DeltaG(GR)(3)). DeltaG(GR)(3) correlate well with DeltaG(GR)(2), and DeltaG(GR)(2) are well analyzed by the Hammett-type dual parameters. DeltaG(GR)(2) and DeltaG(GR)(3) are demonstrated to be controlled by the resonance interaction of the p(Z)-pi(C(6)H(4))-p(Y) conjugations. QC calculations are performed on the ground and exited states of 1 4, which clarify the effective p(Z)-pi(C(6)H(4))-p(Y) conjugations for Z of heavier atoms. PMID- 19558181 TI - Site-isolation and recycling of PdCl(2) using PDMS thimbles. AB - Macroscopic thimbles composed of polydimethylsiloxane (PDMS) were used to site isolate PdCl(2) from the products of Wacker-Tsuji oxidations and Pd-mediated homocouplings. The reactions were completed on the interior of hollow thimbles, and the organic products were isolated by their selective flux to the exterior of the thimbles. Although organic molecules had high flux through the walls of the thimbles, PdCl(2) remained encapsulated as a result of its polar structure that rendered it insoluble in PDMS. In two examples, less than 0.002% of the PdCl(2) added to the interior of the thimble partitioned to the exterior; thus, over 99.998% remained encapsulated on the interior of the thimble. Because it was encapsulated, this catalyst was readily recycled five times for the Wacker-Tsuji oxidation of styrene. A sequential reaction was also completed where p methylstyrene was oxidized to 4'-methyl-acetophenone by PdCl(2) on the interior of a thimble and then fluxed to the exterior to react with phenylmagnesium bromide to yield 1-phenyl-1-p-tolyl-ethanol. This method site-isolated PdCl(2) catalysts without requiring them to be rendered heterogeneous, the addition of exogenous ligands, or any modifications to the catalyst. The catalyst was site isolated by affecting its environment rather than by altering its ligand structure. PMID- 19558182 TI - Synthesis of a core carbon framework of cyanosporasides A and B. AB - Treatment of 3-(2-ethynylphenyl)prop-2-ynyl benzenesulfinate with 2.5 mol % of [RhCl(CO)(2)](2) at 40 degrees C under an atmosphere of CO effected the successive 2,3-sigmatropic rearrangement and carbonylative [2 + 2 + 1] ring closing reaction to afford the 8-(phenylsulfonyl)-1H-cyclopent[a]-inden-2-one in a high yield. Chemical modification of the ring-closed product via lipase mediated optical resolution produced the optically active 3-acetoxy-3a cyclohexyloxy-3,3a-dihydrocyclopent-[a]indene skeleton, the core carbon framework of cyanosporasides A and B. PMID- 19558183 TI - Impact of donor-acceptor geometry and metal chelation on photophysical properties and applications of triarylboranes. AB - Three-coordinate organoboron compounds have recently found a wide range of applications in materials chemistry as nonlinear optical materials, chemical sensors, and emitters for organic light-emitting diodes (OLEDs). These compounds are excellent electron acceptors due to the empty p(pi) orbital on the boron center. When accompanied by electron donors such as amines, these molecules possess large electronic dipoles, which promote donor-acceptor charge-transfer upon excitation with light. Because of this, donor-acceptor triarylboranes are often highly luminescent both in the solid state and in solution. In this Account, we describe our research to develop donor-acceptor triarylboranes as efficient blue emitters for OLEDs. Through the use of hole-transporting donor groups such as 1-napthylphenylamines, we have prepared multifunctional triarylboranes that can act as the emissive, electron transport, or hole transport layers in OLEDs. We have also examined donor-acceptor compounds based on 2,2'-dipyridylamine or 7-azaindolyl donors, several of which have fluorescent quantum efficiencies approaching 100%. We are also investigating the chemistry of metal-containing triarylboranes. Our studies show that the electron-deficient boryl group can greatly facilitate metal-to-ligand charge-transfer transitions and phosphorescence. In addition, electronegative linker groups such as 2,2' bipyridine can act in synergy with metal chelation to greatly improve the electron-accepting ability and Lewis acidity of triarylboranes. Donor-acceptor triarylboranes developed in our laboratory can also serve as a series of "switch on" sensors for fluoride ions. When the donor and acceptor are linked by rigid naphthyl or nonrigid silane linkers, donor-acceptor conjugation is disrupted and charge transfer occurs primarily through space. The binding of fluoride ions to the boron center disrupts this charge transfer, activating alternative pi --> pi* transitions in the molecule and changing the emission color of the sample. More recently, we have used these nonconjugated linkers to prepare organometallic donor-acceptor triarylboranes in which fluorescence and phosphorescence can simultaneously be observed from two different chromophores in the same molecule at ambient temperature. These dual emissive molecules remain sensitive to fluoride ions, and give synergistic singlet-triplet emission responses when titrated with F(-). Fluoride ions can also act as valuable chemical probes, providing insight into the electronic structure of this new class of optoelectronic materials. We have demonstrated that donor-acceptor triarylboranes are promising materials in anion sensing and electroluminescent device applications. Nonetheless, despite our work and that of other research groups, there is still much to be learned about organometallic and multiply emissive triarylboron systems. PMID- 19558184 TI - Identification and quantification of metabolites of orally administered naringenin chalcone in rats. AB - Naringenin chalcone is the main active component of tomato skin extract, which has an antiallergic activity. In this study, naringenin chalcone was orally administered to rats, and the chemical structures and levels of the major metabolites in the plasma and urine of rats were determined. HPLC analysis indicated the presence of three major metabolites in the urine. LC-MS and NMR analyses tentatively identified these as naringenin chalcone-2'-O-beta-D glucuronide, naringenin-7-O-beta-D-glucuronide, and naringenin-4'-O-beta-D glucuronide. Naringenin chalcone-2'-O-beta-D-glucuronide was the only metabolite detected in the plasma, and its peak plasma level was observed 1 h after naringenin chalcone administration. Naringenin chalcone-2'-O-beta-D-glucuronide also inhibited histamine release from rat peritoneal mast cells stimulated with compound 48/80. This activity might contribute to the antiallergic activity of naringenin chalcone in vivo. To the best of the authors' knowledge, this study is the first to report determination of naringenin chalcone metabolites in rat plasma and urine. PMID- 19558185 TI - Efficient charge transport in DNA diblock oligomers. AB - The realization of highly efficient photoinduced charge separation across the pi stacked base pairs in duplex DNA remains elusive. The low efficiencies (<5%) typically observed for charge separation over a dozen or more base pairs are a consequence of slow charge transport and rapid charge recombination. We report here a significant (5-fold or greater) enhancement in the efficiency of charge separation in diblock purine oligomers consisting of two or three adenines followed by several guanines, when compared to oligomers consisting of a single purine or alternating base sequences. This approach to wire-like behavior is attributed to both slower charge recombination and faster charge transport once the charge reaches the G-block in these diblock systems. PMID- 19558186 TI - Inhibition of influenza virus infections by sialylgalactose-binding peptides selected from a phage library. AB - Influenza virus hemagglutinin recognizes sialyloligosaccharides of glycoproteins and glycolipids as cell surface receptors in the initial stage of the infection process. We demonstrate that pentadecapeptides that bind to a sialylgalactose structure (Neu5Ac-Gal) inhibited the infection of cells by influenza virus. The pentadecapeptides were identified through affinity selection from a phage displayed random peptide library using a monolayer of the ganglioside Neu5Acalpha2-3Galbeta1-4Glcbeta1-1'Cer (GM3). The peptides were found to have affinity for GM3, and alanine scanning showed seven amino acid residues that contribute to carbohydrate recognition. The binding of peptides to the cell surface was significantly inhibited in the presence of sialic acid or by the digestion of cell surface sialyl residues by neuraminidase. Plaque assays indicated that a molecular assembly of alkylated peptides inhibited the infection of Madin-Darby canine kidney cells by influenza virus. Carbohydrate-binding peptides that inhibit carbohydrate-virus interaction showed inhibitory activity. These results may lead to a new approach to the design of antiviral drugs. PMID- 19558187 TI - Size limitation in translocation of fibroblast growth factor 1 fusion proteins across the endosomal membrane. AB - After binding to its receptor on the surface of mammalian cells and subsequent endocytosis, FGF1 is translocated across the membrane into the cytosol. The growth factor is then further transported into the nucleus. In order to characterize more closely the translocation mechanism utilized by FGF1, we introduced additional amino acids into FGF1 to test the size dependence of the translocated substrate. We constructed mutants containing an increasing number of copies of the myc tag (1-13 copies) in a surface loop of the FGF1 molecule. All of the constructs bound to specific FGF receptors and to heparin and were taken up by endocytosis. However, only FGF1 mutants harboring up to three myc tags (53 amino acids) were translocated while mutants with five myc tags (77 amino acids) or more were not translocated through the membrane. We further showed that insertion of other, unrelated polypeptides into FGF1, i.e., 3xFLAG tag (22 amino acids) and streptavidin binding peptide (50 amino acids), was also translocated. Larger insertions into FGF1, like the CBP-SBP tag (82 amino acids) or ricin A chain (272 amino acids), resulted in fusion proteins that failed to translocate. The presented data imply that it is possible to employ FGF1 to import various polypeptides into the cytosol and nucleus of cells. Furthermore, the strict size dependence of FGF1 fusion proteins in membrane translocation argues against simple leakage of FGF1 from ruptured endosomal membranes but rather points to a specific translocation apparatus involving a proteinaceous pore. PMID- 19558188 TI - Hepatic OATP1B transporters and nuclear receptors PXR and CAR: interplay, regulation of drug disposition genes, and single nucleotide polymorphisms. AB - Drug uptake transporters are now increasingly recognized as clinically relevant determinants of variable drug responsiveness and unexpected drug-drug interactions. Emerging evidence strongly suggests members of the organic anion transporting polypeptide (OATP) family appear to be particularly important to the disposition of many drugs in clinical use today. Specifically, the liver-enriched OATP1B subfamily members OATP1B1 and OATP1B3 exhibit broad substrate specificity and the ability to transport drugs which are ligands for xenobiotic sensing nuclear receptors such as the pregnane X receptor (PXR) and the constitutive androstane receptor (CAR). Accordingly, OATP1B transporters may indirectly regulate expression of drug metabolism genes via modulation of the intracellular concentration of PXR and CAR ligands. Moreover, a number of functionally important single nucleotide polymorphisms (SNPs) in OATP1B transporters have been described. In this review, a brief summary of known SNPs in PXR and CAR will be followed by an in-depth outline of OATP1B1 and OATP1B3 transporters particularly in relation to the known SNPs in these OATPs and the interplay between OATP1B transporters with PXR and CAR, both in vitro and in vivo. PMID- 19558190 TI - Informing disinvestment through cost-effectiveness modelling: is lack of data a surmountable barrier? AB - The mandatory nature of recommendations made by the National Institute for Health and Clinical Excellence (NICE) in the UK has highlighted inherent difficulties in the process of disinvestment in existing technologies to fund NICE-approved technologies. A lack of evidence on candidate technologies means that the process of disinvestment is subject to greater uncertainty than the investment process, and inefficiencies may occur as a result of the inverse evidence law. This article describes a potential disinvestment scenario and the options for the decision maker, including the conduct of value of information analyses. To illustrate the scenario, an economic evaluation of a disinvestment candidate (screening for amblyopia and strabismus) is presented. Only very limited data were available. The reference case analysis found that screening is not cost effective at currently accepted values of a QALY. However, a small utility decrement due to unilateral vision loss reduced the incremental cost per QALY gained, with screening expected to be extremely cost effective. The discussion highlights the specific options to be considered by decision makers in light of the model-based evaluation. It is shown that the evaluation provides useful information to guide the disinvestment decision, providing a range of focused options with respect to the decision and the decision-making process. A combination of explicit model-based evaluation, and pragmatic and generalizable approaches to interpreting uncertainty in the decision-making process is proposed, which should enable informed decisions around the disinvestment of technologies with weak evidence bases. PMID- 19558189 TI - The G18V CRYGS mutation associated with human cataracts increases gammaS crystallin sensitivity to thermal and chemical stress. AB - GammaS-crystallin, important in maintaining lens transparency, is a monomeric betagamma-crystallin comprising two paired homologous domains, each with two Greek key motifs. An autosomal dominant cortical progressive cataract has been associated with a G18V mutation in human gammaS-crystallin. To investigate the molecular mechanism of this cataract and confirm the causative nature of the G18V mutation, we examined resultant changes in conformation and stability. Human gammaS-crystallin cDNA was cloned into pET-20b(+), and the G18V mutant was generated by site-directed mutagenesis. Recombinant HgammaS-crystallins were expressed in Escherichia coli and purified by ion-exchange and size-exclusion chromatography. By analytical ultracentrifugation wild-type and mutant HgammaS crystallins are monomers of about 21.95 +/- 0.21 and 20.89 +/- 0.18 kDa, respectively, and have similar secondary structures by far-UV CD. In increasing levels of guanidine hydrochloride (GuHCl), a sharp red shift in fluorescence lambda(max) and increase in emission correlating with exposure of tryptophans to the protein surface are detected earlier in the mutant protein. Under thermal stress, the G18V mutant begins to show changes in tryptophan fluorescence above 42 degrees C and shows a Tm of 65 degrees C as monitored by CD at 218 nm, while wild-type HgammaS-crystallin is very stable with Tm values of 75.5 and 75.0 degrees C as measured by fluorescence and CD, respectively. Equilibrium unfolding/refolding experiments as a function of GuHCl confirm the relative instability of the G18V mutant. Wild-type HgammaS-crystallin exhibits a two-state transition and reversible refolding above 1.0 M GuHCl, but the unfolding transition of mutant HgammaS-crystallin shows an intermediate state. The first transition (N --> I) shows a [GuHCl](1/2) of 0.5 M while the second transition (I --> U) has the same [GuHCl](1/2) as wild-type HgammaS-crystallin, about 2.0 M. Our present study confirms the high stability of wild-type HgammaS-crystallin and demonstrates that the G18V mutation destabilizes the protein toward heat and GuHCl-induced unfolding. These biophysical characteristics are consistent with the progressive cataract formation seen in the family members carrying this mutation. PMID- 19558191 TI - Health-promoting schools: evidence for a holistic approach to promoting health and improving health literacy. AB - Chronic diseases are now the major causes of death and disability worldwide, and non-communicable diseases (NCD) account for the majority of the global health burden. About half of premature deaths are related to health-risking behaviours that are often established during youth and extend to adulthood. While these diseases might not be curable, they are preventable. Prevention is possible when sustained actions are directed at individuals and families, as well as at the broader social, economic and cultural determinants of NCD. A 'life-course' approach to promoting healthy behaviour should begin early in life. The aim of this article is to discuss the impact of the 'health-promoting school' (HPS) on improvements in youth health. HPS can be described as a holistic, whole-school approach in which a broad health education curriculum is supported by the environment and ethos of the school. HPS moves beyond individual behavioural change to consider organizational and policy change such as improving the physical and social environment of the school, as well as its curricula and teaching and learning methods. A positive culture for health would facilitate higher levels of health literacy by helping individuals tackle the determinants of health better as they build the personal, cognitive and social skills for maintaining good health. There is reasonable evidence to demonstrate that the whole-school approach using the HPS framework is effective in improving health, ranging from physical activities and healthy eating to emotional health. Schools adopting the HPS framework have demonstrated changes in culture and organizational practice to become more conducive to health improvement. These schools were reported to have better school health policies, higher degrees of community participation, and a more hygienic environment than non-HPS schools, and students in these schools had a more positive health behaviour profile. Health promotion and disease prevention is essential to reduce the healthcare burden of children and adolescents. HPS would help to combat the global burden of childhood obesity by promoting healthy eating behaviours and encouraging higher levels of physical activity. There are gaps in service provision for children and adolescents from both the health and education perspective; the HPS framework has the potential to develop a mechanism of closer integration with the primary healthcare system, making youth health services more school based and student centred. A new model of interconnection between HPS and different components of primary healthcare can be evolved to make services for disease prevention and health promotion more student friendly. PMID- 19558193 TI - Direct economic burden of high-risk and metastatic melanoma in the elderly: evidence from the SEER-Medicare linked database. AB - BACKGROUND: While the clinical implications of advanced melanoma have been extensively documented, little is known about the direct medical costs associated with the disease, particularly for elderly patients who carry the highest disease incidence and morbidity. OBJECTIVES: To document resource utilization and costs to the Medicare system for elderly patients with high-risk (stages IIB/C, IIIA/B, IIIC) or metastatic (stage IV) melanoma. METHODS: Data were taken from the Surveillance, Epidemiology, and End Results (SEER)-Medicare linked database combining clinical information on incident cancer cases in the US between 1991 and 2002 with longitudinal (1991-2005) administrative Medicare claims. Subjects aged > or =65 years with at least one stage IIB or higher melanoma diagnosis were selected. An index date was identified corresponding to the first observed stage IIB or higher diagnosis. Subjects were then categorized into mutually exclusive index disease stages, based on the SEER-reported melanoma stage observed at the index date. All subsequent analyses were stratified according to the index disease stage. Subjects without a record of death were required to have at least 6 months of continuous Medicare Part A and Part B benefits coverage before and after their index date. Subjects who died <6 months after their index date were retained for analysis. Resource utilization and costs were evaluated for each patient from index date until death, benefits cessation or end of the database (31 December 2005). Cost data were inflated to 2007 $US and stratified by the care setting in which they were incurred: inpatient hospital, skilled nursing facility, emergency room, physician office, home healthcare, hospice and other ancillary. RESULTS: 6470 subjects met all inclusion criteria. Index stage distribution was: IIB/C (38%), IIIA/B (46%), IIIC (1%) and IV (15%). Median follow-up was 56, 39, 16 and 6 months, respectively. Patients with stage IV disease had 3.1 hospital days per month, compared with 0.5, 0.6 and 1.1 days for stage IIB/C, IIIA/B and IIIC patients, respectively. Adjusted inpatient costs for stage IV subjects were $US5565 per patient per month versus $US1031, $US1440 and $US2275 for stage IIB/C, IIIA/B and IIIC patients, respectively (p < 0.0001). Adjusted total costs were $US11 471 per month for stage IV subjects, compared with $US2338, $US3395 and $US6885 for stages IIB/C, IIIA/B and IIIC, respectively (p < 0.0001). CONCLUSIONS: The per-patient cost of advanced melanoma is high. Hospital services are the largest component of these costs. Monthly costs for subjects with stage IV melanoma were 67% higher than costs for subjects with stage IIIC disease and >3-fold higher than costs for patients with stages IIIA/B and IIB/C. However, when combining estimated monthly costs with median follow-up duration (a proxy for survival time), total costs incurred by Medicare appear to be highest for patients diagnosed at stage IIIA/B. PMID- 19558192 TI - Cost effectiveness of sirolimus-eluting stents compared with bare metal stents in acute myocardial infarction: insights from the TYPHOON trial. AB - BACKGROUND: Drug-eluting stents have been shown to reduce the rate of repeat revascularization after percutaneous coronary intervention for acute myocardial infarction (AMI) as compared with bare metal stents (BMS). A few studies have reported the cost effectiveness of sirolimus-eluting stents (SES) in several countries, but none in the particular setting of AMI in France. OBJECTIVES: To assess the cost effectiveness of SES compared with BMS in a pre-specified subgroup of French patients with AMI in the randomized, multicentre TYPHOON trial. METHODS: A prospective economic evaluation was conducted for the 337 patients in the TYPHOON trial who were enrolled in the French centres. In the TYPHOON trial, patients with AMI with ST-segment elevation less than 12 hours after the onset of chest pain were randomized to undergo percutaneous coronary intervention with either SES or BMS. Data on clinical outcomes and resource use were collected prospectively over a 1-year follow-up period (from October 2003 to October 2005). Unit costs were applied to the resource utilization data. The main outcome measure was the incremental cost-effectiveness ratio (ICER) for additional cost per target-vessel revascularization (TVR) avoided. The perspective of the study was the French healthcare system and costs were expressed in 2007 values. RESULTS: SES significantly reduced the rate of TVR (6.6% vs 22.2% with BMS, p < 0.0001). There was no difference in the rate of death, recurrent myocardial infarction or stent thrombosis after 1 year of follow up between the SES and BMS groups. Mean index admission costs, including the angioplasty procedure, were increased by 282 Euro per patient in the SES group, mostly driven by the price of the SES. Mean follow-up costs were 140 Euro per patient lower in the SES group. Mean aggregate 1-year costs showed a euro 1142 per-patient increase in the SES group compared with the BMS group. The ICER was 7321 Euro per TVR avoided. CONCLUSIONS: In this pre-specified subgroup analysis of the TYPHOON trial, the use of SES in patients with AMI with ST-segment elevation less than 12 hours after the onset of chest pain reduced the rate of TVR compared with BMS. However, SES had a debatable ICER for the payer if it was based only on the specific benefit of TVR avoided. PMID- 19558194 TI - Oral therapies for the treatment of pulmonary arterial hypertension: a population based cost-minimization analysis. AB - BACKGROUND AND OBJECTIVE: Pulmonary arterial hypertension (PAH) is a rare but life-threatening condition that is characterized by progressive elevation of pulmonary artery pressure and pulmonary vascular resistance, leading to right sided heart failure and frequently death. Orally administered agents used for the treatment of symptomatic, moderate-to-severe PAH include sildenafil and the endothelin (ET) receptor antagonists (ERAs), bosentan and sitaxentan (sitaxsentan). Ambrisentan is a new oral ET(A) receptor-selective ERA, with higher ET receptor affinity than bosentan. Placebo-controlled, randomized clinical trials (RCTs) have demonstrated that ambrisentan (5 or 10 mg/day) is safe and effective. To provide health economic data on the multiple oral PAH therapies currently available, a population-based cost-minimization analysis (CMA) was conducted for Canada. METHODS: The primary requirement for a CMA is that all clinical outcomes be equivalent between comparator treatments. To provide such supporting data, a literature search was conducted for RCTs of oral agents used to treat symptomatic PAH. This was followed by application of direct and indirect statistical methods to support the hypothesis of clinical equivalence between the oral agents. Estimates for PAH prevalence, incidence and death rates were then used to build a population-based CMA model. The base-case analysis considered costs for drug therapy, outpatient pharmacy costs, medical consultations and visits, laboratory and diagnostic procedures and other healthcare-related resources. In addition, costs for secondary pharmacotherapy in cases where the primary agent had to be discontinued because of adverse effects were also included. The time horizon for evaluating pharmacotherapy was 3 years, all costs were in 2008 Canadian dollars ($Can) and the costs were discounted at a rate of 3% annually. The study perspective was the Canadian healthcare system. RESULTS: There were no double-blind RCTs comparing ambrisentan with any of the other oral agents. Therefore, an indirect comparison of placebo-controlled trials of PAH drugs had to be used to support the clinical equivalence. This included a calculation of standardized mean differences (SMD) between agents (vs placebo) and a meta-regression analysis on the primary and secondary trial endpoints. Keeping in mind the caveats associated with indirect trial comparisons, the data suggested similar clinical efficacy over 12-16 weeks between agents, as indicated by the identical magnitude of the SMD between the active agent and placebo and the non-significant differences between drugs as determined by the meta regression analysis. The population-based model projected that the number of PAH patients clinically suitable for these drugs in Canada would be 931 in the first full-budget year (i.e. 2009) with an increase to 1114 by the third full year. The CMA revealed the following rank order of the least to most costly agent; sildenafil, ambrisentan, sitaxentan and bosentan. Sildenafil was the least costly, primarily because of the lower daily drug-acquisition cost. Of the three ERAs, ambrisentan would be associated with annual cost savings of $Can3.4 and $Can5.6 million when used as an alternative to sitaxentan or bosentan, respectively. CONCLUSIONS: Ambrisentan is less costly than other available ERAs, including bosentan and sitaxentan, but is more costly than sildenafil. In PAH patients in whom an ERA is the preferred agent, ambrisentan may be the drug of choice because of its economic advantages and improved safety profile. PMID- 19558195 TI - Development of a budget-impact model to quantify potential cost savings from prescription opioids designed to deter abuse or ease of extraction. AB - BACKGROUND: Prescription opioid (RxO) abuse, dependence and misuse is a significant and growing problem in the US, and is associated with a substantial economic burden. Such abuse may be reduced by theoretical formulations that minimize the possibility of abuse, dependence and misuse of RxOs by injection, snorting, crushing or chewing. As well as public health and safety benefits, use of an abuse-deterrent/resistant RxO (ADO) that may deter abuse, dependence and misuse could also generate cost savings. OBJECTIVES: To estimate potential annual cost savings to US third-party payers realized from introducing a theoretical ADO. METHODS: A budget-impact model (BIM) was developed to assess potential cost savings from the introduction of an ADO to third-party payers. The BIM included information on ADO attributes, costs associated with RxO abuse-related episodes, prevalence of RxO abuse and potential market share capture of the new ADO. Numbers of abuse-related episodes were calculated using a database on admissions to substance abuse treatment centres and other national surveys. Direct (medical and pharmaceutical) costs associated with RxO abuse, dependence and misuse were calculated using de-identified employer claims data (n > 6 000 000) for costs of abuse-related episodes. All cost estimates are in $US, year 2006 values. The BIM was developed for a theoretical prescription drug with therapeutic properties similar to those of controlled-release oxycodone, in a formulation that is specifically designed to resist or deter common methods of abuse, including injection, crushing, snorting and chewing. RESULTS: Potential cost savings to third-party payers from introducing an ADO for the US (assuming a privately insured cost structure) could range from approximately $US0.6 billion to $US1.6 billion per year depending on different possible scenarios. CONCLUSION: While savings estimates from introduction of an ADO depend on a range of assumptions, cost savings would be substantial. PMID- 19558196 TI - Life cycle assessment-driven selection of industrial ecology strategies. AB - The paper presents an application of the Life-Cycle Assessment (LCA) to the planning and environmental management of an "eco-industrial cluster." A feasibility study of industrial symbiosis in southern Italy is carried out, where interlinked companies share subproducts and scraps, services, structures, and plants to reduce the related environmental impact. In particular, the research focuses on new recycling solutions to create open recycling loops in which plastic subproducts and scraps are transferred to external production systems. The main environmental benefits are the reduction of resource depletion, air emissions, and landfilled wastes. The proposed strategies are also economically viable and they suggest cost abatement for the involved companies. This research shows the need for a multidisciplinary approach to data processing and to complexity managing of the investigated systems. In this context, life-cycle thinking is required to be promoted throughout the economy, as well to be as a part of all decisions on products and other criteria such as functionality, health, and safety. The Life-Cycle Assessment approach can be assumed as a methodology for influencing decision makers to make sustainable choices. PMID- 19558197 TI - An approach for assessing cumulative effects in a model river, the Athabasca River basin. AB - Novel approaches addressing aquatic cumulative effects over broad temporal and spatial scales are required to track changes and assist with sustainable watershed management. Cumulative effects assessment (CEA) requires the assessment of changes due to multiple stressors both spatially and temporally. The province of Alberta, Canada, is currently experiencing significant economic growth as well as increasing awareness of water dependencies. There has been an increasing level of industrial, urban, and other land-use related development (pulp and paper mills, oil sands developments, agriculture, and urban development) within the Athabasca River basin. Much of the historical water quantity and quality data for this basin have not been integrated or analyzed from headwaters to mouth, which affects development of a holistic, watershed-scale CEA. The main objectives of this study were 1) to quantify spatial and temporal changes in water quantity and quality over the entire Athabasca River mainstem across historical (1966-1976) and current day (1996-2006) time periods and 2) to evaluate the significance of any changes relative to existing benchmarks (e.g., water quality guidelines). Data were collected from several federal, provincial, and nongovernment sources. A 14% to 30% decrease in discharge was observed during the low flow period in the second time period in the lower 3 river reaches with the greatest decrease occurring at the mouth of the river. Dissolved Na, sulfate, chloride, and total P concentrations in the second time period were greater than, and in some cases double, the 90th percentiles calculated from the first time period in the lower part of the river. Our results show that significant changes have occurred in both water quantity and quality between the historical and current day Athabasca River basin. It is known that, in addition to climatic changes, rivers which undergo increased agricultural, urban, and industrial development can experience significant changes in water quantity and quality due to increased water use, discharge of effluents, and surface run-off. Using the results from this study, we can begin to quantify dominant natural and man-made stressors affecting the Athabasca River basin as well as place the magnitude of any local changes into an appropriate context relative to trends in temporal and spatial variability. PMID- 19558198 TI - Regional reference variation provides ecologically meaningful protection criteria for northern world heritage site. AB - Unprecedented rates of resource development and climate change at northern latitudes coupled with a lack of baseline information limits our ability to set ecologically meaningful criteria needed to protect these inherently sensitive ecosystems. We surveyed water and sediment chemistry, community composition of benthic algae and invertebrates and fish, and condition of a sentinel fish species, slimy sculpin (Cottus cognatus), in 2 rivers adjacent to metal mines and in 20 reference rivers in the headwaters of a World Heritage Site, the South Nahanni River Basin, NWT, Canada. The normal range (i.e., mean +/- 2 standard deviations) of biological conditions in regional reference sites (grouped by community type) were used to set ecologically meaningful effect sizes. These effect sizes were used in noncentral hypotheses tests to assess the ecological condition of potentially impaired sites. Significant impairments at sites influenced by current and historical mining activity were indicative of mild enrichment (e.g., increased benthic abundance and sculpin condition) and bioaccumulation of metals (e.g., increased concentrations of Cu and Fe in muscle tissue of sculpin). Comparisons between our regional reference study and a concurrent upstream-downstream study showed that the sensitivity of biological endpoints was typically related to the impairment criteria used and not to the type of study design. Concentrations of metals such as Al, Cu, and Fe in river water at reference sites were above federal and regional guidelines, suggesting that these guidelines are not appropriate for the metal-rich headwaters of the South Nahanni River. The ephemerellid mayflies Drunella spinifera and Ephemerella maculata were present in 4 of our study sites; their occurrence had not previously been documented in the Yukon or Northwest Territories. Our results confirmed that the lack of baseline information on the physiochemical and biological composition of northern rivers is hampering our ability to evaluate changes in these understudied ecosystems. However, the use of noncentral hypotheses tests based on empirically derived effect sizes enabled us to develop ecologically meaningful protection criteria, maintain statistical rigor, and provide probabilities of impairment that can be used directly in risk assessment. PMID- 19558199 TI - Thresholds of toxicological concern for endocrine active substances in the aquatic environment. AB - The threshold of toxicological concern (TTC) concept proposes that an exposure threshold value can be derived for chemicals, below which no significant risk to human health or the environment is expected. This concept goes further than setting acceptable exposure levels for individual chemicals, because it attempts to set a de minimis value for chemicals, including those of unknown toxicity, by taking the chemical's structure or mode of action (MOA) into consideration. This study examines the use of the TTC concern concept for endocrine active substances (EAS) with an estrogenic MOA. A case study formed the basis for a workshop of regulatory, industry and academic scientists held to discuss the use of the TTC in aquatic environmental risk assessment. The feasibility and acceptability, general advantages and disadvantages, and the specific issues that need to be considered when applying the TTC concept for EAS in risk assessment were addressed. Issues surrounding the statistical approaches used to derive TTCs were also discussed. This study presents discussion points and consensus findings of the workshop. PMID- 19558200 TI - Carcinogenic risk as the basis for fish advisories: a critique. AB - Fish advisories are important tools in public health practice and are primarily used to translate fish contaminant levels into consumption recommendations for consumers. Even when a targeted advisory is issued, it may alter broad food consumption patterns among the public, including diminishing intake of fish-based protein and polyunsaturated fatty acids. Such alterations may have both positive (e.g., reduced exposure to contaminants) and negative (e.g., loss of health benefits or cultural traditions associated with consuming fish) consequences. Currently, a fish advisory may be based on the potential for either noncarcinogenic or carcinogenic endpoints. Consumption recommendations based on a cancer outcome are likely to be highly restrictive, potentially diminishing opportunities for the recognized health benefits associated with a fish-rich diet. This possibility causes us to raise 3 arguments against using cancer risk as the basis for fish consumption advisories. First, the benefits of fish consumption are widely recognized. Second, the standard methodology to predict cancer risk is likely to overestimate actual risk, often by orders of magnitude. Third, the public's real and perceived concerns about cancer may result in unintended consequences, such as avoidance of fish altogether. As an alternative to cancer-based advisories, we suggest that future advisories incorporate a multidisciplinary public health framework focused on avoiding noncarcinogenic health outcomes and encouraging the public to consume a balanced diet rich in fish. We also suggest that decision makers need to 1) understand which elements of the advisory process are science and which are implicit or default policy, 2) consciously consider whether these policy elements are appropriate for their particular situation, and 3) if not, be willing to make and defend alternative policy choices. PMID- 19558201 TI - Recommendations for the development and application of wildlife toxicity reference values. AB - Toxicity reference values (TRVs) are essential in models used in the prediction of the potential for adverse impacts of environmental contaminants to avian and mammalian wildlife; however, issues in their derivation and application continue to result in inconsistent hazard and risk assessments that present a challenge to site managers and regulatory agencies. Currently, the available science does not support several common practices in TRV derivation and application. Key issues include inappropriate use of hazard quotients and the inability to define the probability of adverse outcomes. Other common problems include the continued use of no-observed- and lowest-observed-adverse-effect levels (NOAELs and LOAELs), the use of allometric scaling for interspecific extrapolation of chronic TRVs, inappropriate extrapolation across classes when data are limited, and extrapolation of chronic TRVs from acute data without scientific basis. Recommendations for future TRV derivation focus on using all available qualified toxicity data to include measures of variation associated with those data. This can be achieved by deriving effective dose (EDx)-based TRVs where x refers to an acceptable (as defined in a problem formulation) reduction in endpoint performance relative to the negative control instead of relying on NOAELs and LOAELs. Recommendations for moving past the use of hazard quotients and dealing with the uncertainty in the TRVs are also provided. PMID- 19558202 TI - A risk-ranking methodology for prioritizing historic, potentially contaminated mine sites in British Columbia. AB - The Crown Land Restoration Branch (CLRB) of the British Columbia Ministry of Agriculture and Lands is responsible for managing thousands of historic and abandoned mine sites on provincial lands (referred to as Crown Contaminated Sites). For most of these sites, there is limited information available regarding the extent of potential contamination or potential human health and ecological risks. Given the large number of sites, the CLRB sought a system for prioritizing investigation and management efforts among them. We developed a Risk-Ranking Methodology (RRM) to meet this objective, which was implemented in 2007/2008 with an emphasis on historic mine sites because of the significant number of sites and related potential risk. The RRM uses a risk-based Preliminary Site Investigation to gather key information about the sites. The information for each site is analyzed and summarized according to several attributes aimed at characterizing potential health and ecological risks. The summary information includes, but is not limited to, generic comparisons of exposure with effects levels (screening quotients) for human and ecological exposure pathways. The summary information (more than 25 attributes) is then used in a workshop setting to evaluate relative rankings among sites, and also to identify subsequent management actions for each site. Application of the RRM in 2007/2008 was considered successful, because there was confidence in the process, the content and the outputs. A key challenge was keeping the number of attributes to a manageable level. Ranking was based on discussion and consensus, which was a feasible approach given the relatively small number of sites that need to be ranked each year, and facilitated transparency in the ranking process. We do not rule out the future possibility of developing a quantitative function to capture trade-offs among attributes. PMID- 19558203 TI - Environmental risk assessment of hydrotropes in the United States, Europe, and Australia. AB - An environmental assessment of hydrotropes was conducted under the Organisation for Economic Co-operation and Development (OECD) Screening Information Data Sets (SIDS) for High Production Volume (HPV) Program via the Global International Council of Chemical Associations (ICCA) Hydrotropes Consortium. The assessment and its conclusions were presented at a meeting of the OECD member countries in Washington, DC in 2005. The SIDS Initial Assessment Report (SIAR) was accepted by the membership. Their conclusion was "The chemicals in this [hydrotropes] category are of low priority for further work because of their low hazard profile." Hydrotropes are used to solubilize the water-insoluble ingredients of cleaning and personal care products including, for example, powder and liquid laundry detergents, hard-surface cleaners, machine dishwashing rinse aids, hand dishwashing liquids, body washes, shampoos, hair conditioners, and liquid hand and face soaps. Global production equals approximately 46 500 metric tons, a little more than half of which is used in the United States. The 8 chemicals accounted for in the "hydrotropes category" include ammonium, Ca, K, and Na salts that are described by 10 Chemical Abstract Service (CAS) registration numbers. The 8 chemical entities are generally comparable and predictable in their chemical behavior and that measured and/or modeled data for members from one subgroup can be applied to other subgroups and to the hydrotropes category as a whole. The assessment is based on a search for and evaluation of available data on physical-chemical properties, biodegradability, removal by wastewater treatment, and aquatic toxicity. Reliable ecotoxicity and environmental fate data were found for selected members of the category. Partitioning, once released into the environment, and exposure in surface waters were modeled for consumer use and manufacturing scenarios relevant to the United States, Europe, and Australia. The models indicate 99+% of the hydrotropes will partition to water. Furthermore, given the low potential for hydrotropes reaching the terrestrial environment and their lack of persistence or bioaccumulation, the focus of the assessment is on the aquatic environment, specifically the water compartment. Aquatic risks were assessed in each scenario using what is referred to as the PEC/PNEC ratio. The modeled predicted environmental concentration (PEC), accounting for volume released and wastewater treatment, is divided by the predicted no effect concentration (PNEC) derived from the aquatic toxicity tests. The closest a predicted environmental concentration came to the toxicity threshold is 0.125 (or 12.5% of the no effect concentration) and that is for a hypothetical manufacturing facility that produces the entire annual volume of hydrotropes and discharges to a small (10%ile) stream under low flow (7Q10) conditions. PEC/PNEC ratios were considerably smaller for consumer use scenarios. The ratios were 0.0002 for a low flow (7Q10) stream scenario in the United States, 0.026 to 0.089 for regional and local water bodies, respectively, in Europe, and 0.004 to 0.036 for oceans and rivers, respectively, in Australia. In conclusion, aquatic hazard levels are not expected to be reached under exaggerated conditions of manufacture or normal consumer use of hydrotropes. Hydrotropes are neither persistent nor bioaccumulative in the environment. PMID- 19558205 TI - Environmental fate factors and human intake fractions for risk assessment of petroleum products. AB - Petroleum products may contain up to thousands of individual hydrocarbon compounds, which vary widely in environmental behavior and ecotoxicity. Environmental risk assessment of these complex substances is facilitated by use of the hydrocarbon block method (HBM). The HBM assigns similarly behaving hydrocarbons to the same blocks and then relates release rates of petroleum products to environmental concentrations and human intake rates by means of so called environmental fate factors and human intake fractions of the hydrocarbon blocks. We have derived such fate factors and intake fractions and associated uncertainties with a library of representative hydrocarbon structures. Fate factors and intake fractions of over 1500 individual representative hydrocarbons have been modeled according to the EU Technical Guidance Documents. Fate factors and intake fractions for the chosen hydrocarbon blocks are then estimated from average values obtained for the individual compounds in the blocks. Fate factors and intake fractions of the hydrocarbon blocks vary by up to 10 orders of magnitude for the different emission scenarios and compartments/spatial scales. The highest fate factors are obtained for the exposure scenarios with the highest emission intensity (the local emission scenarios) and for the most direct exposure chain. Uncertainties introduced by the "blocking" method are typically smaller than 30% and rarely larger than a factor of 3. Various hydrocarbon blocking schemes of different resolution were tested, and all schemes investigated appeared to perform satisfactorily. The use of the library-based HBM was illustrated by assessing the aquatic ecological risk of gas oil, emitted to water. The aquatic risk characterization ratio of gas oil hydrocarbons in regional freshwater is estimated to be 6 * 10(-5) per unit of emission (1 kg/d released 1:10:100 to local, regional, and continental scales, respectively), with an uncertainty of 32%. In view of other uncertainties in environmental risk assessment, the precision achieved with the procedure presented here is judged to be reasonable. PMID- 19558206 TI - Using data from drug discovery and development to aid the aquatic environmental risk assessment of human pharmaceuticals: concepts, considerations, and challenges. AB - Over recent years, human pharmaceuticals have been detected in the aquatic environment. This, combined with the fact that many are (by design) biologically active compounds, has raised concern about potential impacts in wildlife species. This concern was realized with two high-profile cases of unforeseen environmental impact (i.e., estrogens and diclofenac), which have led to a flurry of work addressing how best to predict such effects in the future. One area in which considerable research effort has been made, partially in response to regulatory requirements, has been on the potential use of preclinical and clinical pharmacological and toxicological data (generated during drug development from nonhuman mammals and humans) to predict possible effects in nontarget, environmentally relevant species: so-called read across. This approach is strengthened by the fact that many physiological systems are conserved between mammals and certain environmentally relevant species. Consequently, knowledge of how a pharmaceutical works (the "mode-of-action," or MoA) in nonclinical species and humans could assist in the selection of appropriate test species, study designs, and endpoints, in an approach referred to as "intelligent testing." Here we outline the data available from the human drug development process and suggest how this might be used to design a testing strategy best suited to the specific characteristics of the drug in question. In addition, we review published data that support this type of approach, discuss the potential pitfalls associated with read across, and identify knowledge gaps that require filling to ensure accuracy in the extrapolation of data from preclinical and clinical studies, for use in the environmental risk assessment of human pharmaceuticals. PMID- 19558207 TI - Effects of topical administration of nimodipine on cerebral blood flow following subarachnoid hemorrhage in pigs. AB - We sought to explore whether topical administration of nimodipine improves the abnormal cerebral perfusion following subarachnoid hemorrhage (SAH) in pigs. Fourteen pigs were randomly divided into three groups: sham (n=4), SAH (n=5), or SAH + nimodipine (n=5). The SAH model was established by injecting fresh autologous nonheparinized arterial blood into the suprasellae cistern. Nimodipine or saline placebo (0.04 g/mL) were administered to the operative area on the fourth day after the SAH model was established. The cerebral blood flow (CBF) was measured 60 min after topical administration of nimodipine by cranial SPECT/CT scans with 5 mCi 99mTc-ECD injected intravenously. The CCR (corticocebellar ratio) was calculated by dividing the counts/voxel of the whole cerebral hemisphere by the average count/voxel in the cerebellar region of reference and RD (relative dispersion). A predictor for impaired autoregulation of CBF was calculated by dividing standard deviation (SD) of regional perfusion by mean perfusion (RD=SD/Mean). CCR and RD were applied to describe hemisphere CBF and perfusion heterogeneity. Cerebral perfusion significantly decreased in the SAH group (CCR: 1.382+/-0.192, RD: 0.417+/-0.015) compared to sham (CCR: 1.988+/ 0.346, RD 0.389+/-0.015) (p<0.05). Abnormal cerebral perfusion status, however, was not significantly improved in the nimodipine + SAH group (CCR: 1.503+/-0.107, RD: 0.425+/-0.018) compared to the SAH group (p>0.05). Topical administration of nimodipine did not significantly improve CBF following SAH. These findings were not consistent with our previous data demonstrating that the topical administration of nimodipine significantly alleviates cerebral vasospasm following SAH detected by TCD. Potential mechanisms governing these disparate outcomes require further investigation. PMID- 19558208 TI - Prior ethanol injection promotes brain edema after traumatic brain injury. AB - Alcohol consumption prior to traumatic brain injury (TBI) promotes morbidity and mortality although the mechanisms involved remain unclear. The morbidity and mortality caused by TBI, especially brain contusion, are known to be closely associated with brain edema. Here we examined the effects of ethanol pretreatment on brain edema, inflammatory responses, and oxidative stress after brain contusion. Male Wistar rats were given 3 g/kg ethanol intraperitoneally and 1 h later were subjected to brain contusion. The ethanol-pretreated group had a significantly decreased survival rate. Magnetic resonance imaging showed ethanol pretreatment significantly augmented the volume of cytotoxic brain edema after contusion. In the ethanol-pretreated rat, the activities of NF-kappaB and AP-1 were reduced 6 h after contusion and COX-2 mRNA expression was increased 24 h after contusion. These findings suggest that ethanol augmented cerebral edema and mortality in rats with brain contusion, possibly through actions on cell survival pathways or COX-2 expression. In addition, antioxidant treatment at 3 h post injury significantly attenuated some markers of oxidative stress, mortality, and volume of edema at 24 h after ethanol treatment and contusion. PMID- 19558209 TI - Hydrogen peroxide as a cell-survival signaling molecule. AB - Reactive oxygen species (ROS) were seen as destructive molecules, but recently, they have been shown also to act as second messengers in varying intracellular signaling pathways. This review concentrates on hydrogen peroxide (H2O2), as it is a more stable ROS, and delineates its role as a survival molecule. In the first part, the production of H2O2 through the NADPH oxidase (Nox) family is investigated. Through careful examination of Nox proteins and their regulation, it is determined how they respond to stress and how this can be prosurvival rather than prodeath. The pathways on which H2O2 acts to enable its prosurvival function are then examined in greater detail. The main survival pathways are kinase driven, and oxidation of cysteines in the active sites of various phosphatases can thus regulate those survival pathways. Regulation of transcription factors such as p53, NF-kappaB, and AP-1 also are reviewed. Finally, prodeath proteins such as caspases could be directly inhibited through their cysteine residues. A better understanding of the prosurvival role of H2O2 in cells, from the why and how it is generated to the various molecules it can affect, will allow more precise targeting of therapeutics to this pathway. PMID- 19558210 TI - Initiative for a new diabetes therapeutic approach in a Mediterranean country: the INDEED study. AB - AIM: To assess the efficacy of a strategy to improve vascular risk management in patients with type 2 diabetes mellitus (T2DM). METHODS: This was a pilot best practice implementation enhancement programme that enrolled 578 patients with T2DM. A baseline visit was followed by a concerted effort from previously trained physicians to improve adherence to lifestyle advice and optimise drug treatment for all vascular risk factors. The patients were followed-up for 6 months. The UKPDS risk engine was used to estimate vascular risk in patients without established coronary heart disease (CHD) (n = 279). RESULTS: There was an improvement in compliance to lifestyle measures and increased prescription of evidence-based medication. In patients without established CHD there was a 37% reduction in estimated risk for CHD, 44% for fatal CHD, 10% for stroke and 25% for fatal stroke (p < or = 0.003 for all comparisons vs. baseline). There was also a substantial increase in the proportion of patients with established CHD who achieved their vascular risk factor targets. CONCLUSIONS: This is the first study to increase the adherence to multiple interventions in patients with T2DM in both primary care and hospital settings. Education of physicians and patients, distribution of guidelines/brochures, and the completion of a one-page form, motivated both physicians and patients to achieve multiple vascular risk factor goals. PMID- 19558211 TI - Nitric oxide in cell survival: a janus molecule. AB - Nitric oxide (NO), plays multiple roles in the nervous system. In addition to regulating proliferation, survival and differentiation of neurons, NO is involved in synaptic activity, neural plasticity, and memory function. Nitric oxide promotes survival and differentiation of neural cells and exerts long-lasting effects through regulation of transcription factors and modulation of gene expression. Signaling by reactive nitrogen species is carried out mainly by targeted modifications of critical cysteine residues in proteins, including S nitrosylation and S-oxidation, as well as by lipid nitration. NO and other reactive nitrogen species are also involved in neuroinflammation and neurodegeneration, such as in Alzheimer disease, amyotrophic lateral sclerosis, Parkinson disease, multiple sclerosis, Friedreich ataxia, and Huntington disease. Susceptibility to NO and peroxynitrite exposure may depend on factors such as the intracellular reduced glutathione and cellular stress resistance signaling pathways. Thus, neurons, in contrast to astrocytes, appear particularly vulnerable to the effects of nitrosative stress. This article reviews the current understanding of the cytotoxic versus cytoprotective effects of NO in the central nervous system, highlighting the Janus-faced properties of this small molecule. The significance of NO in redox signaling and modulation of the adaptive cellular stress responses and its exciting future perspectives also are discussed. PMID- 19558213 TI - Impact of interaction of polymorphic forms of p53 codon 72 and N-acetylation gene (NAT2) on the risk of lung cancer in the North Indian population. AB - The interaction of genetic and environmental factors can determine individual susceptibility to various cancers. We studied the influence of NAT2 and codon 72 p53 polymorphisms on 151 patients with lung cancer and an equal number of matched population controls. Polymorphisms of NAT2 and p53 were determined by PCR-RFLP techniques. The results were analyzed using logistic regression analysis. A statistically significant relationship between NAT2*5 and NAT2*6 alleles and lung cancer risk was observed. In addition, the population with slow acetylator alleles for NAT2*5 and NAT2*6 had a significantly higher risk of lung cancer compared with rapid acetylator alleles both in smokers and nonsmokers. The combined genotype of heterozygous arginine (Arg)/proline (Pro), Pro/Pro, and slow acetylator alleles of NAT2*5 and NAT2*6 showed higher, although not significant, risk of lung cancer compared with Arg/Arg and rapid acetylator alleles of NAT2*5 and NAT2*6. In conclusion, these findings suggest that the influence of NAT2 genotype, alone or in combination with p53 genotype, may confer increased susceptibility to lung cancer. PMID- 19558212 TI - Mitochondrial glutathione, a key survival antioxidant. AB - Mitochondria are the primary intracellular site of oxygen consumption and the major source of reactive oxygen species (ROS), most of them originating from the mitochondrial respiratory chain. Among the arsenal of antioxidants and detoxifying enzymes existing in mitochondria, mitochondrial glutathione (mGSH) emerges as the main line of defense for the maintenance of the appropriate mitochondrial redox environment to avoid or repair oxidative modifications leading to mitochondrial dysfunction and cell death. mGSH importance is based not only on its abundance, but also on its versatility to counteract hydrogen peroxide, lipid hydroperoxides, or xenobiotics, mainly as a cofactor of enzymes such as glutathione peroxidase or glutathione-S-transferase (GST). Many death inducing stimuli interact with mitochondria, causing oxidative stress; in addition, numerous pathologies are characterized by a consistent decrease in mGSH levels, which may sensitize to additional insults. From the evaluation of mGSH influence on different pathologic settings such as hypoxia, ischemia/reperfusion injury, aging, liver diseases, and neurologic disorders, it is becoming evident that it has an important role in the pathophysiology and biomedical strategies aimed to boost mGSH levels. PMID- 19558214 TI - Uptake of foreign nucleic acids in kidney tubular epithelial cells deficient in proapoptotic endonucleases. AB - Degradation of DNA during gene delivery is an obstacle for gene transfer and for gene therapy. DNases play a major role in degrading foreign DNA. However, which of the DNases are involved and whether their inactivation can improve gene delivery have not been studied. We have recently identified deoxyribonuclease I (DNase I) and endonuclease G (EndoG) as the major degradative enzymes in the mouse kidney proximal tubule epithelial (TKPTS) cells. In this study, we used immortalized mouse TKPTS cells and primary tubular epithelial cells isolated from DNase I or EndoG knockout (KO) mice and examined the degradation of plasmid DNA during its uptake. DNase I and EndoG KO cells showed a higher rate of transfection by pECFP-N1 plasmid than wild-type cells. In addition, EndoG KO cells prevented the uptake of fluorescent-labeled RNA. Complete inhibition of secreted DNase I by G-actin did not improve plasmid transfection, indicating that only intracellular DNase I affects DNA stability. Data demonstrate the importance of DNase I and EndoG in host cell defense against gene and RNA delivery to renal tubular epithelial cells in vitro. PMID- 19558215 TI - Transforming growth factor-beta1 up-regulation of human alpha(1)(I) collagen is mediated by Sp1 and Smad2 transacting factors. AB - Hepatic fibrosis results from excessive deposition of type I collagen. The roles of Smads in mediating the effect of transforming growth factor-beta1 (TGFbeta1) on activation of the alpha(1)(I) collagen promoter were determined. Smads bind in association with Sp1 to the CC(GG)-rich TGFbeta1 responsive element of the promoter that lacks the classical Smad recognition element, and enhance binding of Sp1. In transfection experiments, TGFbeta1 activated a proximal promoter, but not promoters mutated at sites that prevented Sp1 binding. Sp1 alone or the combination of Smad2 and Smad4 activated the promoter in transfected human LX-2 stellate cells. Sp1 or Smad2 knockdowns with siRNAs prevented the effect of TGFbeta1 in enhancing the promoter. In conclusion, this study shows that Smads bind in association with Sp1 to the CC(GG)-rich TGFbeta1 responsive element of the human alpha(1)(I) collagen promoter that lacks the classical Smad recognition element, thus enhancing the binding of Sp1 and in this manner activating the collagen promoter. PMID- 19558216 TI - A combination of proatherogenic single-nucleotide polymorphisms is associated with increased risk of coronary artery disease and myocardial infarction in Asian Indians. AB - Common single-nucleotide polymorphisms (SNPs) in genes of lipid metabolism modestly influence plasma low-density lipoprotein cholesterol (LDL-C) and risk of coronary artery disease (CAD). We evaluated a panel of LDL-C-modulating SNPs for potential association with risk of CAD in Asian Indians. Fifteen SNPs of CETP, ABCB1, APOAI, CYP7A1, and HMGCR genes were genotyped in 265 CAD patients and 150 controls of North Indian origin. A proatherogenic genotype score was formulated based on number of alleles associated with LDL-C and was evaluated for association with risk of CAD. We observed 12 SNPs from CETP, APOAI, ABCB1, CYP7A1, and HMGCR genes to be associated with baseline LDL-C and high-density lipoprotein cholesterol levels and increased risk of CAD (p < 0.05). Co occurrence of three or more risk alleles (proartherogenic genotype score >or=3) was associated with increased risk of CAD and myocardial infarction. Analysis of epistatic interactions revealed CETPTaqIB1B1/405II/APOAI-75GA to be best model of CAD risk prediction in our population. Our study highlights synergistic association of multiple SNPs of lipid pathway with LDL-C levels and risk of CAD, and indicates that co-occurrence of proatherogenic risk alleles may provide incremental information about CAD risk beyond lipid concentrations. PMID- 19558217 TI - In vitro study on DNA binding of ruthenium(II) complexes with polypyridyl ligands. AB - A new ligand {2-(5-nitro-furan-2-yl)-1H-1,3,7,8-tetraaza-cyclopenta[l] phenanthrene} (nftp) and its Ru(II) complexes [Ru(phen)(2)(nftp)](2+) (1) (phen = 1,10-phenanthroline) and [Ru(bpy)(2)(nftp)](2+) (2) (bpy = 2,2'-bipyridine) were synthesized and characterized. The binding properties of the two complexes to calf thymus-DNA (CT-DNA) were investigated by different spectrophotometric methods and viscosity measurements together with equilibrium dialysis and circular dichroism spectroscopy. The results suggest that both complexes bind to DNA through intercalation and enantioselectively interact with CT-DNA. However, complex 1 is a better candidate as an enantioselective binder to CT-DNA than complex 2. Although no emission is generally observed in water or organic solvents for Ru(II) polypyridyl complexes with a nitro group, complexes 1 and 2 can emit luminescence in both media. When irradiated at 365 nm, complex 1 cleaves DNA more effectively than complex 2. PMID- 19558218 TI - A comparative study on the interaction of cis- and trans-platin with DNA and RNA. AB - Cis-diamminedichloroplatinum(II) is a frequently used and very effective chemotherapeutic drug for treatment of various malignancies; however, the trans isomer is clinically ineffective. Cis-platin exerts its antitumor activity by binding to DNA via intrastrand cross-links to d(GpG) (dG = deoxyguanosine) and to d(ApG) (dA = deoxyadenosine), interfering with DNA replication and transcription and causing cell death. The trans-diamminedichloroplatinum(II) isomer also binds DNA, but is clinically ineffective. This study was designed to examine the interactions of cis- and trans-platin with calf thymus DNA and yeast RNA in aqueous solution at physiological conditions, using a constant DNA and RNA concentration (6.25 mM) and various platin salts/polynucleotide (phosphate) ratios of 1/100, 1/50, 1/25, and 1/12.5. Fourier transform infrared, ultraviolet visible spectroscopic methods were used to determine the drug binding modes, the binding constants, and the stability of cis- and trans-platin-DNA and -RNA complexes in aqueous solution. Spectroscopic evidence showed that cis- and trans platin bind to the major and minor grooves of DNA (via G, A, T, and C bases), while RNA binding is through G, U, A, and C bases with some degree of the pt phosphate (PO(2)) interaction for both isomers and overall binding constants of K((cis-platin-DNA)) = 5.51 x 10(4) M(-1), K((trans-platin-DNA)) = 2.26 x 10(4) M( 1), K((cis-platin-RNA)) = 1.9 x 10(4) M(-1), and K((trans-platin-RNA)) = 1.75 x 10(4) M(-1). DNA and RNA aggregations occurred at high platin concentrations. No biopolymer conformational changes were observed upon cis- and trans-platin interactions, while DNA remains in the B-family, and RNA retains its A-family structure. The order of platin compound-polymer stability was cis-platin-DNA > trans-platin-DNA > cis-platin-RNA > trans-platin-RNA. PMID- 19558219 TI - Generation of induced pluripotent stem cells by efficient reprogramming of adult bone marrow cells. AB - Reprogramming of somatic cells provides potential for the generation of specific cell types, which could be a key step in the study and treatment of human diseases. In vitro reprogramming of somatic cells into a pluripotent embryonic stem (ES) cell-like state has been reported by retroviral transduction of murine fibroblasts using four embryonic transcription factors or through cell fusion of somatic and pluripotent stem cells. Here we show that mouse adult bone marrow mononuclear cells (BM MNCs) are competent as donor cells and can be reprogrammed into pluripotent ES cell-like cells. We isolated BM MNCs and mouse embryonic fibroblasts (MEFs) from Oct4-GFP transgenic mice, fused them with ES cells, or infected them with retroviruses expressing Oct4, Sox2, Klf4, and c-Myc. Fused BM MNCs formed more ES-like colonies than did MEFs. Infected BM MNCs gave rise to induced pluripotent stem (iPS) cells, although transduction efficiencies were not high. It was more efficient to pick up iPS colonies as compared with MEFs. BM derived iPS (BM iPS) cells expressed ES cell markers, formed teratomas, and contributed to chimera mice with germ line development. Clonal analysis revealed that BM iPS clones had diversity, although some clones were found to be genetically identical with different phenotypes. Our findings imply that BM MNCs have potential advantages to generate iPS cells for the clinical application. PMID- 19558221 TI - Seeding of pancreatic islets into prevascularized tissue engineering chambers. AB - Transplantation of islets into the portal vein of diabetic patients has emerged as a promising procedure for the treatment of type 1 diabetes. However, shortages of donors and adverse effects leading to graft impairment and/or rejection have prevented this procedure from achieving widespread clinical application. The aim of this study was to develop a method that could support the survival and function of transplanted islets using a prevascularized tissue engineering chamber. Islets were transplanted into tissue engineering chambers established on the epigastric pedicle in the groin of diabetic mice. Islets were transplanted at the time of chamber implantation or with 21 days prevascularization of the chamber. Transplantation of islets into prevascularized chambers into diabetic RIP-K(b) mice resulted in a significant reduction in blood glucose levels that became evident in the third week and improved glycemic control as measured by a glucose tolerance test. This study highlights that islet survival and function are potentiated by allowing a period of prevascularization within tissue engineering chambers before islet transplantation. This novel prevascularized chamber may be an improved method of islet transplantation. It can be easily accessed for islet seeding, easily retrieved, and transplanted to alternative anatomical sites by microvascular methods. PMID- 19558220 TI - Tracking chondrocytes and assessing their proliferation with carboxyfluorescein diacetate succinimidyl ester: effects on cell functions. AB - Distinguishing between implanted and host-derived cells, as well as cells of different phenotypes, is important in determining mechanisms of cell-based repair of cartilage. The objectives of this study were to assess the utility of carboxyfluorescein diacetate, succinimidyl ester ("CFDA, SE" or CFSE) for tracking chondrocytes from superficial (S) and middle (M) zones and their proliferation, and to determine the effects of CFSE on the chondrocyte functions, proliferation, and synthesis of proteoglycan 4 (PRG4) and glycosaminoglycan (GAG). CFSE-labeled and unlabeled S and M zone chondrocytes were plated in either low- or high-density (10,000 or 200,000 cells/cm(2)) monolayer, incubated, and analyzed on days 1 and 7. Cell suspensions were analyzed for retention of CFSE by flow cytometry and fluorescence microscopy and for cell proliferation by assay for DNA and GAG. Cultures were also analyzed for newly synthesized PRG4. Deconvolution of flow cytometric histograms was done to determine the number of cells in each doubling generation. Most chondrocytes were labeled consistently and intensely labeled with CFSE through 10 cycles of division. At day 7 of culture, approximately 95% of S and M zone cells seeded at high density could be distinguished as fluorescent. Chondrocyte proliferation and synthesis of PRG4 were unaffected by cell labeling, while GAG synthesis was slightly diminished. CFSE may be useful in determining the fate and function of implanted chondrocytes in vivo. PMID- 19558222 TI - The Liss maneuver: a nonendoscopic technique for difficult Foley catheterization. AB - PURPOSE: Difficult male catheterization often results in the need for bedside cystoscopy; herein, we describe a simple maneuver that may preclude the need for a more involved and expensive endoscopic procedure. MATERIALS AND METHODS: A 0.035-inch Amplatz Super Stiff guidewire was placed floppy end first, down an 18F Foley catheter lumen; the guidewire was then clamped proximally to the body of the catheter at its point of exit. To test the added stiffness that the wire imparted to the tip and body of the catheter, different size Bardex I.C. Foley catheters were tested with a 0.035-inch 145-cm Amplatz Super Stiff guidewire placed in an Chattillon Force Gauge (TCM 201 Asset #4665) to measure bending force and tip strength in pounds of pressure. RESULTS: The catheter body bending force (stiffness) was increased on average 359%, but the tip of the catheter, holding the floppy end of the guidewire, showed no significant increase in stiffness. A trend was seen showing larger percent differences in stiffness for smaller catheters (12F and 14F); indeed, the guidewire shaft stiffness approached the same stiffness as the shaft of a standard 20F catheter, but the stiffness of the catheter's tip remained unchanged. Clinically, the guidewire stiffening technique was used successfully in five of six patients; the failure was in a patient with a hitherto undiagnosed 8F urethral stricture. CONCLUSION: In cases of difficult bladder catheter placement, the Super Stiff guidewire technique can be used to increase the stiffness of the catheter to navigate the torturous urethra with any size catheter before pursuing other more invasive and time consuming techniques. If this maneuver fails, the same guidewire can be used in conjunction with the flexible endoscope to then place a Councill catheter. PMID- 19558223 TI - Acid and organic aerosol coatings on magnetic nanoparticles increase iron concentrations in human airway epithelial cells. AB - Numerous industrial applications for man-made nanoparticles have been proposed. Interactions of nanoparticles with agents in the atmosphere may impact human health. We tested the postulate that in vitro exposures of respiratory epithelial cells to airborne magnetic nanoparticles (MNP; Fe(3)O(4)) with and without a secondary organic aerosol (SOA) and an inorganic acid could affect iron homeostasis, oxidative stress, and interleukin (IL)-8 release. Cell iron concentrations were increased after exposures to MNP and values were further elevated with co-exposures to either SOA or inorganic acid. Increased expression of ferritin and elevated levels of RNA for DMT1, proteins for iron storage and transport respectively, followed MNP exposures, but values were significant for only those with co-exposures to inorganic acid and organic aerosols. Cell iron concentration corresponded to a measure of oxidative stress in the airway epithelial cells; MNP with co-exposures to SOA and inorganic acid increased both available metal and indices of oxidant generation. Finally, the release of a proinflammatory cytokine (i.e. IL-8) by the exposed cells similarly increased with cell iron concentration. We conclude that MNP can interact with a SOA and an inorganic acid to present metal in a catalytically reactive state to cultured respiratory cells. This produces an oxidative stress to affect a release of IL-8. PMID- 19558224 TI - Biological effect of carbon graphite whisker in rat lung by long-term Inhalation. AB - Carbon graphite whisker (CGW) was used in a 1-year inhalation study in male Wistar rats and its biological effect was observed until the 1-year clearance period. The inhalation study was conducted at 2.6 +/- 0.5 mg/m(3) (equivalent to 44.5 +/- 15.0 fibers/mL) for 6 hours a day, 5 days a week for 1 year. There were no differences in survival rate between the exposure and control groups during this examination; however, the body weights were significantly different at the end of the 1-year clearance. The lung weight at 3 days and 1 year after the end of exposure was not significantly different in both groups. The deposited amount of CGW was 6.83 +/- 0.75 mg at 3 days post-exposure; the deposition rate was 17.6%. Only around 30% of the total deposited CGW was cleared during the 1-year clearance period. The geometric means of CGW in the lung, i.e. CMD (count median diameter) and CML (count median length), hardly changed, and the clearance was delayed. In the histopathological examination, there was mild fibrosis in all exposed rats irrespective of the clearance period. One adenoma was observed in a single animal at 3 days post-exposure, while no adenomas were observed in the exposure group after the 1-year clearance. Epithelial hyperplasia was found in some animals. PMID- 19558225 TI - Occupational exposure to bacterial single cell protein induces inflammation in lung and blood. AB - Bacterial single cell protein (BSCP) is used as a protein enrichment in livestock and fish feed, and is extracted from dried bacterial mass. In the production of BSCP, workers are exposed to organic dust containing high levels of endotoxins that may induce acute airway inflammation. However, the long term effect on the airways of such exposure is not known, and we have examined inflammatory markers in induced sputum and blood among BSCP exposed workers. We included 21 non smoking production workers (age 31-42 (range; mean 35)) without respiratory symptoms and 21 healthy non-exposed references (age 21-52 (range; mean 34)). Airborne endotoxin concentrations were measured, and induced sputum samples and blood samples were collected from the workers and non-exposed references. The airborne endotoxin concentration measured in inhaled air during the work shift was 430 EU/m(3) (50-2000) (median (range)). The percentage of neutrophils in induced sputum was 79% (66-93) (median (25th-75th percentiles)) and 31% (25-45) (p < 0.001) for operators and references, respectively. Protein analysis in induced sputum supernatant showed significantly elevated levels of interleukins IL-1beta and IL-12 (p < 0.05), while blood analysis showed significantly elevated levels of PDGF-BB (platelet-derived growth factor-BB) and RANTES (regulated upon activation normally T cell expressed and secreted) (p < 0.05). Workers exposed to BSCP had an airway inflammation characterized by a high level of neutrophils. However, only a few cytokines were elevated in lung and blood, which could imply low inflammatory activity suggestive of possible adaptation mechanisms due to daily exposure to BSCP, or that the inflammation reaction was a dose-related response occurring at higher levels. PMID- 19558226 TI - Proceedings from the 11th International Inhalation Symposium, June 11-14, 2008. PMID- 19558227 TI - European and international standardisation progress in the field of engineered nanoparticles. AB - CEN-STAR supports researchers to be experts in standardization committees in new challenging issues: Standardization needs to reach consensus from new knowledge. In Europe, pre-normative and co-normative research can be supported financially by the FP7 for research. The area of nanoscience and nanotechnology is developing new standards in Europe within the CEN/TC 352, at the international level at the ISO/TC 229. Particular works are about measurement methods and tests for toxicity at the nanoscale. Some activities are done within the Organisation for Economic Cooperation and Development (OECD) and in some other parts of the world, such as the Asian Nano-Forum and the National Institute of Standards and Technology (NIST) in the United States. Main items for standardization include: definition terminology; metrology instrumentation, reference methods, and materials; measurement methods for applications (physical, chemical, material, electronics); and measurement methods for interface in health, environment, and safety. For toxicity at the nanoscale, particular issues have to be resolved, such as: reproducibility of experiments; reference protocols and use of quality management standards for research (ISO 9001); and intercomparability of results following "round robin" tests. The little information about standardization in nanotechnology in Europe includes the Code of Conduct for Responsible Nanosciences and Nanotechnologies Research and the European Commission mandate to CEN/TC 352, as well as the European Union (EU) projects NanoStrand, NanoInteract, NanoImpactNet, and others. PMID- 19558228 TI - Dissolution behaviour of a nanoparticle in a microscale volume of solvent: thermodynamic and kinetic considerations. AB - The dissolution behaviour of an oxidic nanoparticle in a small volume of solvent was investigated. The results of thermodynamic and kinetic calculations are presented. Variations in nanoparticle size and solvent volume are considered. Two kinds of final states of the system can be formed. One state is a homogeneous monomolecular solution of the dissolved species; the other one is a system in stable equilibrium between nanoparticle and solvent. What kind of state is formed depends on the size of the nanoparticle and the amount of solvent. The concentration of the dissolved species is much higher than the saturation concentration of the bulk material in many cases. An unusual dissolution behaviour of an ensemble of nanoparticles, called kinetic size effect, follows from the calculations. A very high concentration is found of the dissolved material at the beginning of the dissolution process. The concentration decreases at longer dissolution times. As an example, the experimental results of the dissolution kinetics of an ensemble of nanoparticles of commercial titanium dioxide in water are presented. Good agreement between experimental results and theoretical calculations is found. From these data it can be deduced that a nanoparticle with a radius of 14.15 nm dissolves in a volume of water of 18 microm(3) until its radius becomes 13.77 nm and a stable system nanoparticle dissolved substance-solvent is formed (supersaturation 24), whereas a nanoparticle of radius 11.4 nm is completely dissolved after 73 h (supersaturation 165). The effect is established for different oxides. PMID- 19558229 TI - Facing the key workplace challenge: assessing and preventing exposure to nanoparticles at source. AB - Nanomaterials present new challenges to understanding, predicting, and managing potential health risks in occupational environments. In this study, we characterize the key physical processes related to formation and growth of nanoparticles. The main focus is on various occupational environments, as these are known to be major environments with nanoparticles in indoor air. The protection of people potentially to be exposed to nanoparticles is one of the key issues in terms of risk assessment and prevention. Two of the main protection techniques that are discussed and characterized are ventilation and filtration, which are widely used in practical applications. PMID- 19558230 TI - Workplace exposure to engineered nanoparticles. AB - Measurement strategies as well as results from measurements at workplaces are presented. Measurements were performed in research laboratories as well as by companies producing small amounts of nanostructured materials. The work was done in the frame of a BAuA (Bundesanstalt fur Arbeitsschutz und Arbeitsmedizin) research project. Measurement results during manufacturing and processing of TiO(2), nanofibers as well as synthetic ceramic nanopowders and nanostructured materials used for the electrical industry are demonstrated. Measurements were conducted with a scanning mobility particle sizer (Grimm) and a concentration particle counter (TSI). Mean number concentrations of nanoparticles are presented. During determination of particle number concentrations, a distinction must be drawn between the particle number in the ambient air and the particle number resulting from work processes. Thus a special measurement strategy was developed including measurement of the ambient air and also analysis by electron microscopy to define the size of agglomerates as well as the data related to the chemical composition. To date, no significant increase of nanoparticle number has been found provided proper handling was guaranteed (e.g., closed systems, extractor hood). Nevertheless, a few particle agglomerates of the materials used during the work process, as shown by electron microscopy, were detected. Probably nanostructured materials can primarily be released during cleaning and maintenance operations as well as in the case of a failure of normal operation. PMID- 19558231 TI - Charge-based personal aerosol samplers. AB - There are several good reasons to use personal monitors for exposure control and health effect studies. But current personal monitoring methods are either not sensitive enough to measure typical ambient concentrations, work offline (masking short exposures to high concentrations), and/or require trained personnel to analyze the data, which makes them difficult to use. For this reason, we propose the use of a diffusion charging sensor as an online personal monitoring method, and present a miniaturized device (45 x 80 x 200 mm, 770 g) that works on this principle. Our device has a high time resolution and covers typically encountered ambient concentration ranges. It can measure very low particle concentrations of a few hundred particles per cubic centimeter even for ultrafine particles (i.e., two to three orders of magnitude more sensitive than rival technologies), while the upper detection limit is 1 million particles/cm(3), which hardly ever occurs in ambient settings. While other methods measure a fixed quantity, the response of our device can be tuned to be proportional to the particle diameter to the power of x, with at least 0.3 or=110 mg/dL, blood pressure >or=130/>or=85, triglyceride >or=150 mg/dL, high-density lipoprotein cholesterol (HDL-C) <40 mg/dL (men) or <50 mg/dL (women), but body mass index (BMI) >or=30 in lieu of waist circumference >102 cm (40 inches) for men and 88 cm (35 inches) for women. We also accepted current pharmacotherapy for diabetes as qualifying for elevated fasting blood sugar (FBS); current therapy with niacin, gemfibrozil, or fenofibrate for elevated triglyceride concentrations; and recent use of multiple International Classification of Diseases, 9(th) Revision (ICD-9) codes for hypertension for elevated blood pressure. RESULTS: We examined all clinical records for veterans registered in VANCHCS who filled any prescription between July 1, 2004, and June 30, 2005 (n = 51,026). Their average age was 63 years; 93% were male. In all 25% (n = 13,010) were diagnosed as having metabolic syndrome by meeting at least 3 of the above 5 criteria. Because only 60% (n = 30,727) of the population had data for 3 or more criteria, the actual percent with metabolic syndrome is probably substantially higher. CONCLUSIONS: Over one quarter of veterans in the VANCHCS may have metabolic syndrome based on our modified ATP III criteria. We urge screening more veterans with fasting laboratory testing. Computerized screening of a large clinical database can provide an effective strategy to aid clinicians in identifying more patients at risk for cardiovascular disease. PMID- 19558267 TI - Health care utilization and costs by metabolic syndrome risk factors. AB - BACKGROUND: This study compared prevalent health utilization and costs for persons with and without metabolic syndrome and investigated the independent associations of the various factors that make up metabolic syndrome. METHODS: Subjects were enrollees of three health plans who had all clinical measurements (blood pressure, fasting plasma glucose, body mass index, triglycerides, and high density lipoprotein cholesterol) necessary to determine metabolic syndrome risk factors over the 2-year study period (n = 170,648). We used clinical values, International Classification of Diseases, Ninth Revision (ICD-9) diagnoses, and medication dispensings to identify risk factors. We report unadjusted mean annual utilization and modeled mean annual costs adjusting for age, sex, and co morbidity. RESULTS: Subjects with metabolic syndrome (n = 98,091) had higher utilization and costs compared to subjects with no metabolic syndrome (n = 72,557) overall, and when stratified by diabetes (P < 0.001). Average annual total costs between subjects with metabolic syndrome versus no metabolic syndrome differed by a magnitude of 1.6 overall ($5,732 vs. $3,581), and a magnitude of 1.3 when stratified by diabetes (diabetes, $7,896 vs. $6,038; no diabetes, $4,476 vs. $3,422). Overall, total costs increased by an average of 24% per additional risk factor (P < 0.001). Costs and utilization differed by risk factor clusters, but the more prevalent clusters were not necessarily the most costly. Costs for subjects with diabetes plus weight risk, dyslipidemia, and hypertension were almost double the costs for subjects with prediabetes plus similar risk factors ($8,067 vs. $4,638). CONCLUSIONS: Metabolic syndrome, number of risk factors, and specific combinations of risk factors are markers for high utilization and costs among patients receiving medical care. Diabetes and certain risk clusters are major drivers of utilization and costs. PMID- 19558268 TI - Continuous positive airway pressure therapy improves hypoadiponectinemia in severe obese men with obstructive sleep apnea without changes in insulin resistance. AB - BACKGROUND: Obstructive sleep apnea (OSA) is associated with several conditions that could facilitate the onset of cardiovascular and metabolic dysfunctions. Continuous positive airway pressure (CPAP) therapy has been shown to improve cardiovascular morbidity and mortality related to OSA, but the mechanisms underlying this association are not fully understood. OBJECTIVE: The aim of the present study was to evaluate whether sleep apnea contributes to insulin resistance and inflammatory marker alterations and to evaluate the benefits of nasal CPAP therapy in severe obese patients with OSA. METHODS: Plasma inflammatory cytokines and the homeostasis model assessment of insulin resistance index (HOMA-IR, Insulin Sensitivity Index [ISI]) were measured in severe obese male with OSA (n = 16) and compared with body mass index (BMI)-matched male controls without OSA (n = 13). Seven patients with severe sleep apnea (apnea hypopnea index >30 events/h) were reevaluated after 3 months of nasal CPAP therapy. RESULTS: OSA patients had a significantly lower adiponectin levels than obese controls (8.7 +/- 1.18 ng/mL vs. 15.0 +/- 2.55 ng/mL, P = 0.025). HOMA-IR, ISI, tumor necrosis factor-alpha (TNF-alpha, C-reactive protein (CRP), and interleukin-6 (IL-6) levels were not different between groups. Although insulin resistance index and BMI values did not change after 3 months of nCPAP therapy, adiponectin levels increased (P = 0.036) and the levels of TNF-alpha tended to decrease (P = 0.065). Changes in adiponectin levels during nCPAP therapy were positively correlated with an improvement in minimum oxygen saturation (r = 0.773; P = 0.041) and negatively correlated with changes in TNF-alpha levels (r = -0.885; P = 0.008). CONCLUSIONS: nCPAP therapy reverses hypoadiponectinemia levels present in obese men with OSA, probably through reductions in hypoxia and inflammation activity. PMID- 19558269 TI - Polymorphism of Pro12Ala in the peroxisome proliferator-activated receptor gamma2 gene in Iranian diabetic and obese subjects. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma2 (PPARgamma2) is a nuclear receptor that regulates adipocyte differentiation, lipid metabolism, and insulin sensitivity. The aim of this study was to investigate the association between the Pro12Ala single nucleotide polymorphism (SNP) at the PPARgamma2 gene and type II diabetes (T2DM) and obesity in an Iranian population. METHODS: The genomic DNA of the 312 subjects included four groups: (1) nonobese with type II diabetes, (2) obese without type II diabetes, (3) obese with type II diabetes, and (4) nondiabetic nonobese controls. The Pro12Ala polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: Frequencies of the Ala allele in obese subjects were significantly different from those control subjects (odds ratio [OR], 2.358; 95% confidence interval [CI], 1.101-5.05) (P = 0.025). In contrast, no significant association was detected between the Pro12Ala polymorphism and type II diabetes (OR, 0.652; 95% CI, 0.261-1.628). In all subjects, the Ala carriers had a higher body mass index (BMI) compared with the common allele. CONCLUSIONS: Our results showed that the Pro12Ala polymorphism in the PPARgamma2 gene is associated with obesity in Iranian subjects and the presence of the Ala allele could predict higher BMI. PMID- 19558270 TI - Effectiveness and safety of 1-year ad libitum consumption of a high-catechin beverage under nutritional guidance. AB - BACKGROUND: It has been reported that a continuous intake of a catechin beverage will reduce body fat. Traditionally, improvement of eating and exercise habits has been the basis for prevention and reduction of obesity. In this study, we conducted a trial involving human subjects who ingested a catechin beverage for 1 year under nutritional guidance. METHODS: This study was conducted based on a comprehensive cohort design using a catechin beverage (containing 588 mg of tea catechins) and a control beverage (containing 126 mg of tea catechins). At both the start and the end of the trial, the subjects underwent an annual health check and computer tomography for measurement of their abdominal fat. In addition, a food intake survey was conducted and all subjects were provided nutritional guidance by a registered dietitian every 3 months. RESULTS: Data were analyzed using per protocol samples of 134 subjects (catechin group, n = 77; control group, n = 57). Body weight and body mass index were reduced significantly in the catechin group compared to the control group. Changes in body weight during the study period were -1.1 kg in the catechin group and 0.2 kg in the control group. In the catechin group, the visceral fat areas at the start of the trial were significantly correlated with the magnitude of fat reduction at the end of the trial. Under the guidance of a registered dietitian, subjects in the catechin group who showed a reduction in their fat-derived energy percentage during the test period tended to reduce more body weight than those with an increase in this percentage, although no difference in total energy intake was noted between the two groups. One-year ad libitum consumption of a catechin beverage posed no health risks and resulted in a reduction in body weight. CONCLUSIONS: An overall improvement in dietary habits might enhance the weight-reduction effect of the beverage. PMID- 19558271 TI - Chronic kidney disease has a more powerful impact on peripheral arterial disease than metabolic syndrome in Japanese type 2 diabetic patients. AB - BACKGROUND: Chronic kidney disease (CKD) and metabolic syndrome have been recognized as risk factors for cardiovascular disease. However, there is no information comparing their impact on macroangiopathy in diabetic patients. Thus, we studied the prevalence of CKD and metabolic syndrome in Japanese type 2 diabetic patients and then compared their impact on peripheral arterial disease (PAD) in type 2 diabetic patients. METHODS: This study focused on Japanese type 2 diabetic patients without hemodialysis (n = 1014). Patients with albuminuria, including microalbuminuria and/or an estimated glomerular filtration rate less than 60 mL/min/1.73(2), were diagnosed as having CKD. PAD was defined as ankle brachial blood pressure index less than 0.9. RESULTS: The prevalence of CKD and metabolic syndrome was 47.1% and 39.6%, respectively. In four age- and duration matched groups classified by the presence or absence of CKD and metabolic syndrome, the prevalence of PAD was significantly higher in groups with CKD alone than those with metabolic syndrome alone, and the high prevalence in the groups with CKD was not influenced by the coexistence with metabolic syndrome. CONCLUSIONS: This study indicates that CKD has more powerful impact on PAD than metabolic syndrome in type 2 diabetic patients. PMID- 19558273 TI - Index of central obesity is better than waist circumference in defining metabolic syndrome. AB - OBJECTIVE: The International Diabetes Federation (IDF) global definition of metabolic syndrome suggests using race- and gender-specific waist circumference (WC) cutoffs. Previously, we have hypothesized that need for gender- and race specific cutoffs could be obviated by supplanting WC with index of central obesity (ICO). The aim of this study was to test the utility of ICO in defining metabolic syndrome. MATERIALS AND METHODOLOGY: Data were collected from National Health and Nutrition Examination Survey (NHANES) database for year 2005-2006. Subjects were analyzed for presence of metabolic syndrome using National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) criteria. The IDF definition was modified by replacing WC with ICO. Sensitivity and specificity of the IDF definition and modified definition were compared against the NCEP ATP III definition. RESULTS: Using a modified IDF definition, a common cutoff of 0.53 could be obtained for both males and females. The modified IDF definition improved sensitivity from 0.85 to 0.98 and 0.98 to 0.99 among males and females, respectively. This was at the cost of compromised specificity, which reduced from 1.0 to 0.89 and 0.98 for males and females, respectively. CONCLUSIONS: ICO appears better than WC in defining metabolic syndrome in terms of better sensitivity and potential to be used with single cutoff for both genders. PMID- 19558274 TI - Effectiveness of a multidisciplinary program for management of obesity: the Unite d'Enseignement, de Traitement et de Recherche sur l'Obesite (UETRO) database study. AB - BACKGROUND: Obesity is a worldwide health problem assuming epidemic proportions. Development of effective clinical interventions is needed to lower the impact of associated morbidity and mortality, without forgetting related costs. We have established an interdisciplinary clinic for obesity management, Unite d'Enseignement, de Traitement et de Recherche sur l'Obesite (UETRO), which consists of individual consultations combined with group sessions. We report here the effectiveness of this program for weight reduction over the first year of follow up. METHODS: We performed retrospective analysis of standardized patient records of the first 115 consecutive subjects referred to UETRO with available follow up for 1 year. RESULTS: Mean age, body mass index (BMI), and waist circumference (WC) of our cohort were 46 +/- 13 years, 44.7 +/- 0.9 kg/m(2), and 120.5 +/- 1.9 cm, respectively. Hypertension and diabetes were present in 46% and 23% of our patients. Weight and WC loss were gradual over 1 year and were significantly reduced by 6.6 +/- 0.8 kg and 6.7 +/- 0.7 cm, respectively (P < 0.001), without attainment of a plateau. Blood pressure and lipid profile significantly improved after 1 year of follow up. However, the proportion of patients taking metformin, lipid-lowering, antihypertensive, or antiobesity drugs increased significantly over follow up, reflecting intensification of treatment of co-morbidities and weight management. Significant weight and WC loss occurred independently of diabetes status and use of antiobesity medications. CONCLUSIONS: This program appears to be as effective for treating obesity as more intensive treatment programs. Future prospective studies are needed to evaluate the benefits and costs of this therapeutic approach. PMID- 19558272 TI - Metabolic syndrome and the burden of cardiovascular disease in Caribbean Hispanic women living in northern Manhattan: a red flag for education. AB - BACKGROUND: Metabolic syndrome has the highest prevalence among Mexican-American women. Little information is available for Caribbean Hispanics, the largest and fastest growing ethnic minority in the United States. We sought to evaluate the frequency of metabolic syndrome and its relationship with race/ethnicity, socioeconomic position, and education in women of largely Caribbean Hispanic origin. METHODS: There were 204 women enrolled in a cross-sectional study who had demographics, fasting glucose, lipid profile, waist circumference, and blood pressure determined. Metabolic syndrome (defined by the National Cholesterol Education Program/Adult Treatment Panel III [NCEP/ATP III]) was analyzed using univariate and multivariate logistic regression to test age, race/ethnicity, education, health insurance, and residence on the risk of metabolic syndrome. A P value <0.05 was considered significant. RESULTS: Mean age was 58 +/- 11 years, Hispanic 44.1% (93% Caribbean), non-Hispanic white (NHW) 38.7%, and non-Hispanic black 9.8%. Education was some high school ( or = 15 micromol/L. RESULTS: Mean plasma Hcy level of adolescents with metabolic syndrome was found to be 11.8 +/- 5.0 micromol/L. Although the Hcy level of females (11.4 +/- 5.5 micromol/L) was lower than that of males (12.0 +/- 4.7 micromol/L), the difference was not statistically significant (P > 0.05). In all, 40.5% of adolescents had Hcy levels between 10 and 15 micromol/L (mild HH) and 17.7% of adolescents had Hcy levels of > or = 15 micromol/L. The Hcy level was negative and weakly correlated with insulin resistance in females (r = -0.319, P = 0.058) and negative strongly correlated with SBP in males (r = - 0.385, P = 0.011). There were no associations between Hcy level and other components of metabolic syndrome. CONCLUSIONS: An elevated plasma Hcy level is not considered to be related to components of metabolic syndrome. But HH may be an independent risk factor, especially for diabetic adolescents or those who have cardiovascular events as in adults. PMID- 19558277 TI - The coagulation cascade and surgical hemostasis. Introduction. PMID- 19558276 TI - Alterations in blood-brain barrier permeability to large and small molecules and leukocyte accumulation after traumatic brain injury: effects of post-traumatic hypothermia. AB - We investigated the temporal and regional profile of blood-brain barrier (BBB) permeability to both large and small molecules after moderate fluid percussion (FP) brain injury in rats and determined the effects of post-traumatic modest hypothermia (33 degrees C/4 h) on these vascular perturbations. The visible tracers biotin-dextrin-amine 3000 (BDA-3K, 3 kDa) and horseradish peroxidase (HRP, 44 kDa) were injected intravenously at 4 h or 3 or 7 days post-TBI. At 30 min after the tracer infusion, both small and large molecular weight tracers were detected in the contusion area as well as remote regions adjacent to the injury epicenter in both cortical and hippocampal structures. In areas adjacent to the contusion site, increased permeability to the small molecular weight tracer (BDA 3K) was evident at 4 h post-TBI and remained visible after 7 days survival. In contrast, the larger tracer molecule (HRP) appeared in these remote areas at acute permeable sites but was not detected at later post-traumatic time periods. A regionally specific relationship was documented at 3 days between the late occurring permeability changes observed with BDA-3K and the accumulation of CD68 positive macrophages. Mild hypothermia initiated 30 min after TBI reduced permeability to both large and small tracers and the infiltration of CD68 positive cells. These results indicate that moderate brain injury produces temperature-sensitive acute, as well as more long-lasting vascular perturbations associated with secondary injury mechanisms. PMID- 19558278 TI - Achieving hemostasis in the surgical field. AB - Hemostatic mechanisms are an integral part of the human physiology. Traditionally divided into intrinsic and extrinsic arms, the coagulation cascade converges, through the interactions of many different factors, at a common element-thrombin. As a consequence, a number of different agents have been developed to supplement this common, critical step to aid surgical hemostasis. Intraoperative interventions most commonly include sutures and heat-generating cautery devices; however, these methods are sometimes insufficient or inappropriate for a specific procedure or anatomic location, leading to the development of other adjunctive therapies, including topical hemostats. Topical hemostatic agents generally act as active, passive, and combinations therapies, depending on their individual composition and mode of action. We provide a brief review of the normal coagulation cascade, including critical points, followed by a discussion of surgical strategies and adjuctive therapies used to achieve surgical hemostasis and concluding with a discussion of topical thrombins. PMID- 19558279 TI - Topical thrombins: benefits and risks. AB - Whether from surgical misadventure, inherent patient factors, or iatrogenic causes, postoperative bleeding can be a consequence of any surgical procedure. There are many methods and products available to assist in managing or preventing bleeding. For each method, there may be specific benefits and indications, but they may also carry some degree of risk. Topical thrombin is used extensively in many surgical specialties, especially in the cardiovascular and neurosurgical arenas where other hemostatic modalities may not be appropriate choices. As a class, topical thrombins are generally a safe and effective method for achieving intraoperative hemostasis; however, some members of the class carry associated risks with their use. For example, the United States Food and Drug Administration required the addition of a black-box warning to the prescribing information of bovine-derived thrombin, the oldest member of the class, due to concerns of immune-mediated coagulopathies developing after use. In addition, human thrombin derived from pooled plasma has its own, if theoretical, risk of transmitting infections due to viral or prion agents. We address the topical thrombin class and review each product in the context of the current literature. PMID- 19558280 TI - Immune-mediated coagulopathy: a case report. AB - Surgical hemostasis may be achieved by using a number of physical, chemical, or biologic methods. One such method is with topical thrombin; however, one member of that class of drugs, bovine-derived thrombin, is associated with potentially serious consequences such as development of immune-mediated coagulopathy. This case report describes a 61-year-old man with peripheral artery disease who presented with a nonhealing ulcer between his toes. Previous exposure to bovine thrombin was unknown but was considered likely because of his extensive surgical history that included procedures in which topical thrombin is commonly used. The patient was admitted and underwent lower extremity revascularization during which he received his first documented exposure to bovine-derived thrombin. By postoperative day 9, he developed a 2.7-cm retroperitoneal hematoma that had progressed to 9.6 cm by postoperative day 13. Evacuation of the hematoma was performed, during which the patient received his second known exposure to topical bovine thrombin. Based on a plasma mixing study on postoperative day 25, presence of factor V and thrombin inhibitors was suspected. A hematology consultation determined that the patient had developed an immune-mediated coagulopathy manifested as exaggerated laboratory coagulation values that continued even after discontinuation of oral anticoagulation, treatment with multiple transfusions of fresh frozen plasma, and intravenous vitamin K administration. The patient was discharged, after no further bleeding episodes had occurred, on postoperative day 29. Although determining previous exposure to bovine-derived thrombin or presence of antibodies can be difficult, a surgeon's index of suspicion should be raised in patients experiencing coagulopathy if they have previously undergone vascular, cardiac, or spinal procedures in which they were most likely exposed to topical thrombin. PMID- 19558281 TI - Thrombin products: economic impact of immune-mediated coagulopathies and practical formulary considerations. AB - Thrombin has demonstrated utility in aiding surgical hemostasis since its introduction more than 60 years ago. It is used across a wide variety of surgical procedures by virtually every specialty. Only recently have new equally effective and safe products entered the market, causing decision makers to evaluate formulary selection among products with otherwise modest differences. This evaluation includes identifying costs beyond those of acquisition and storage, as well as indirect factors such as monitoring or specialized distribution requirements. One factor to consider specifically in selection of topical thrombin products is the potential for patients to develop an immune-mediated coagulopathy (IMC) after exposure to bovine-derived thrombin. Costs due to adverse drug events fall into the category of indirect costs and, in some instances, can be substantial if bleeding due to IMC occurs. PMID- 19558283 TI - The regenerative capacity of the notochordal cell: tissue constructs generated in vitro under hypoxic conditions. AB - OBJECT: The intervertebral disc (IVD) is a highly avascular structure that is occupied by highly specialized cells (nucleus pulposus [NP] cells) that have adapted to survive within an O(2) concentration of 2-5%. The object of this study was to investigate the effects of long-term hypoxic and normoxic tissue cultures of nonchondrodystrophic canine notochordal cells-cells that appear to protect the disc NP from degenerative change. METHODS: The authors obtained notochordal cells from nonchondrodystrophic canines according to their established methods and placed them into monolayer and 3D culture using sodium alginate globules under either hypoxic (3.5% O(2)) or normoxic (21% O(2)) conditions. Histological, immunohistochemical, scanning electron microscopy, and histomorphometric methods were used to evaluate the cells within the globules after 5 months in culture. RESULTS: Notochordal cells under in vitro hypoxic tissue culture conditions produced a highly complex, organized, 3D cellular construct that was strikingly similar to that observed in vivo. In contrast, traditional normoxic tissue culture conditions resulted in notochordal cells that failed to produce an organized matrix. Hypoxia resulted in a matrix rich in aggrecan and collagen II, whereas normoxic cultured cells did not produce any observable aggrecan or collagen II after 5 months of culture. CONCLUSIONS: Hypoxia induces notochordal cells to organize a complex 3D cellular/extracellular matrix without an external scaffold other than suspension within sodium alginate. These cells produce an extracellular matrix and large construct that shares exactly the same characteristics as the in vivo condition-robust aggrecan, and type II collagen production. Normoxic tissue culture conditions, however, lead to a failure of these cells to thrive and a lack of extracellular matrix production and significantly smaller cells. The authors suggest that future studies of NP cells and, in particular, notochordal cells should utilize hypoxic tissue culture conditions to derive meaningful, biologically relevant conclusions concerning possible biological/molecular interventions. PMID- 19558282 TI - An orthotopic murine model of human spinal metastasis: histological and functional correlations. AB - OBJECT: There is currently no reproducible animal model of human spinal metastasis that allows for laboratory study of the human disease. Consequently, the authors sought to develop an orthotopic model of spinal metastasis by using a human lung cancer cell line, and to correlate neurological decline with tumor growth. METHODS: To establish a model of spinal metastasis, the authors used a transperitoneal surgical approach to implant PC-14 lung tumors into the L-3 vertebral body of nude mice via a drill hole. In 24 animals, motor function was scored daily by using the validated semiquantitative Basso-Beattie-Bresnahan (BBB) scale. A second group of 26 animals (6 or 7 per time point) were sacrificed at specific times, and the spines were removed, sectioned, and stained. Canal compromise was analyzed quantitatively by determining the ratio of the area of the neural elements to the area of the spinal canal on histological sections (neural/canal ratio). Correlations between BBB score and histological evaluation of tumor growth were assessed. RESULTS: Lung cancer xenografts grew in all animals undergoing functional evaluation (24 mice) according to a reliable and reproducible time course, with paraplegia occurring at a median interval of 30 days following tumor implantation (95% CI 28.1-31.9 days). Importantly, the analysis defined 4 key milestones based on components of the BBB score; these were observed in all animals, were consistent, and correlated with histological progression of tumor. From Days 1 to 14, the mean BBB score declined from 21 to 19. The animals progressed from normal walking with the tail up to walking with the tail constantly touching the ground (milestone 1). The median time to tail dragging was 12 days (95% CI 10.8-13.2). Histological studies on Day 14 demonstrated that tumor had progressed from partial to complete VB infiltration, with initial compression of the neural elements and epidural tumor extension to adjacent levels (mean neural/canal ratio 0.32 +/- 0.05, 7 mice). From Days 15 to 20/21 (left/right leg), the mean BBB score declined from 19 to 14. Animals showed gait deterioration, with the development of dorsal stepping (milestone 2). The median time to dorsal stepping was 21 days (95% CI 19.4-22.6) in the left hindlimb and 23 days (95% CI 20.6-25.4) in the right hindlimb. Histological studies on Day 21 demonstrated an increase in the severity of the neural element compression, with tumor extending to adjacent epidural and osseous levels (mean neural/canal ratio 0.19 +/- 0.05, 6 mice). From Days 22 to 26/27 (left/right leg), the mean BBB score declined from 14 to 8. Animals had progressive difficulty ambulating, to the point where they showed only sweeping movements of the hindlimb (milestone 3). The median time to hindlimb sweeping was 26 days (95% CI 23.6-28.4) and 28 days (95% CI 27.1-28.9) in the left and right hindlimbs, respectively. Histological studies on Day 28 revealed progressive obliteration of the spinal canal (mean neural/canal ratio 0.09 +/- 0.01, 7 mice). From Days 29 to 36, the animals progressed to paralysis (milestone 4). The median time to paralysis was 29 days (95% CI 27.6-30.4) and 30 days (95% CI 28.1-31.9) in the left and right hindlimbs, respectively. CONCLUSIONS: The authors have developed an orthotopic murine model of human spinal metastasis in which neurological decline reproducibly correlates with severity of tumor progression. Although developed for lung cancer, this model can be expanded to study other types of metastatic or primary spinal tumors. Ultimately, this will allow testing of targeted therapies against specific tumor types. PMID- 19558284 TI - Developing consistently reproducible intervertebral disc degeneration at rat caudal spine by using needle puncture. AB - OBJECT: The goal in this study was to develop a convenient, less-invasive animal model to monitor progression of intervertebral disc (IVD) degeneration for future testing of new treatments for disc degeneration. METHODS: Level 5/6 and 7/8 IVDs of rat caudal spine were stabbed laterally with 18- or 21-gauge hypodermic needles to a depth of 5 mm from the subcutaneous surface with the aid of fluoroscopy. In vivo MR imaging studies were performed at 4, 8, and 12 weeks postsurgery to monitor progression of IVD degeneration. Histological analysis including H & E and safranin O staining, and immunohistochemical studies of collagen type II and bone morphogenetic protein receptor type II (BMPRII) were assessed at 12 weeks postsurgery. RESULTS: The 18- and 21-gauge needle-stabbed discs illustrated decreases in both the T2 density and MR imaging index starting at 4 weeks, with no evidence of spontaneous recovery by 12 weeks. Histological staining demonstrated a decreased nucleus pulposus (NP) area, and the NP-anulus fibrosus border became unclear during the progression of disc degeneration. Similar patterns of degenerative signs were also shown in both safranin O- and collagen type II-stained sections. The BMPRII immunohistochemical analysis of stabbed discs demonstrated an increase in BMPRII expression in the remaining NP cells and became stronger in anulus fibrosus with the severity of disc degeneration. CONCLUSIONS: After introducing an 18- or 21-gauge needle into the NP area of discs in the rat tail, the stabbed disc showed signs of degeneration in terms of MR imaging and histological outcome measurements. Changes in BMPRII expression in this animal model provide an insight for the effectiveness of delivering BMPs into the region responsible for chondrogenesis for disc repair. This convenient, less-invasive, reproducible, and cost-effective model may be a useful choice for testing novel treatments for disc degeneration. PMID- 19558285 TI - Surgery in the cervicothoracic junction with an anterior low suprasternal approach alone or combined with manubriotomy and sternotomy: an approach selection method based on the cervicothoracic angle. AB - OBJECT: The authors propose an easy MR imaging method to measure and categorize individual anatomical variations within the cervicothoracic junction (CTJ). Furthermore, they propose guidelines for selection of the appropriate approach based on this new categorization system. METHODS: In the midsagittal section of the cervicothoracic MR imaging studies obtained in 95 Chinese patients, a triangle was drawn among 3 points: the suprasternal notch (SSN), the midpoint of the anterior border of the C7/T1 intervertebral disc, and the corresponding anterior border in the CTJ at the level of the SSN. The angle above the SSN was specified as the cervicothoracic angle (CTA). The spatial position between the brachiocephalic vein (BCV), the aortic arch, and the CTA was also measured. Based on these measurements involving the CTA, 3 different patient-specific categorizations are proposed to assist surgeons with selection of the appropriate anterior approach to the CTJ. Three categories of operative approach based on whether the most caudal part of the lesion site was above, within, or below the area of the CTA were classified. The patients were divided into long- or short necked groups based on whether their own CTA was greater than (long necked) or less than (short necked) the average CTA. Finally, a left BCV was called superiorly located when it coursed above the manubrium. The method was evaluated in 21 patients with spinal bone tumors in the CTJ to illustrate the measurement of both the CTA and the great vessels, and corresponding approach selections. RESULTS: In this series of 95 patients, the most common vertebra above the SSN was T-3, especially the upper one-third of T-3. The mean CTA was 47.64 degrees . The left BCV was superior to the manubrium in 21.1% of the 95 cases, and 93.6% of the left BCVs were at the T-2 and T-3 levels. Type A and most Type B lesions could be addressed via a low suprasternal approach, or this approach combined with manubriotomy, if necessary. Type C lesions falling below the CTA will need alternative exposure techniques, including manubriotomy, sternotomy, lateral extracavitary, or thoracotomy. The spinal levels that could be exposed in the long-necked CTJ group were always 1 or 2 vertebral levels lower than those in the short-necked CTJ group during the anterior low suprasternal approach without the manubriotomy. CONCLUSIONS: Imaging of the thoracic manubrium should be routinely included on MR imaging studies obtained in the CTJ. It is important for the surgeon to understand the pertinent anatomy of the individual patients and to determine the feasible surgical approaches after evaluating the CTA and vascular factors preoperatively. An anterior low suprasternal approach, or this approach combined with manubriotomy, is applicable in most of the cases in the CTJ. It should be cautioned that preoperatively unrecognized variations of the left BCV above the SSN might result in potential intraoperative trauma during an anterior approach. PMID- 19558286 TI - Conservative treatment of upper cervical spine injuries with the halo vest: an appropriate option for all patients independent of their age? AB - OBJECT: Most upper cervical spine injuries are able to heal conservatively by halo vest application. The acceptance of the halo is different among patients due to the weight of the apparatus and the limited mobility it causes. Additionally, the fracture healing rate in older patients seems to be inferior to that in younger patients, which would make an operative stabilization procedure more appropriate for the elderly. Furthermore, the risk of complications is assumed to be higher in older people. The purpose of this retrospective study was to find out if there are differences in the clinical and radiological results and in complication rates between 2 patient groups with upper cervical spine injuries and halo vest treatment with special interest of their age group. METHODS: A total of 29 patients with upper cervical spine injuries were treated by halo vest application. The clinical and radiological results and the complication and revision surgery rates were identified. Patients were divided into 2 groups (18 patients were < 65 years and 11 were > 65 years), and the results were evaluated to examine whether there were any differences. RESULTS: The clinical and radiological results and the complication rate were not statistically significantly different between the 2 patient groups; however, there was a tendency for a longer time interval for fracture healing and more complications in the elderly people. CONCLUSIONS: If the conditions for conservative treatment of upper cervical spine injuries with halo fixation are right, the clinical and radiological results are good and almost similar in patients regardless of their age, although there is a tendency for more complications in older people. PMID- 19558287 TI - Postoperative displacement of hydroxyapatite spacers implanted during double-door laminoplasty. AB - OBJECT: Double-door laminoplasty using hydroxyapatite (HA) spacers has been widely performed for compressive cervical myelopathy and has provided good neurological outcome. Although HA spacers are used for preventing reclosure of the opened laminae, they are often displaced or dislocated from their original position. The authors investigated the incidence and patterns of postoperative HA spacer displacement to determine the reasons for this unfavorable event. METHODS: Eighty-six patients with compressive myelopathy underwent double-door laminoplasty in which a total of 278 HA spacers were used. The displacement of HA spacers and opened laminae were assessed using postoperative lateral radiographs and CT scans. RESULTS: Postoperative dorsal migration > 2 mm was found in 116 (42%) of 278 implanted HA spacers. In addition, 33 (38%) of 86 HA spacers rotated > 10 degrees and 29 (34%) of the 86 opened laminae tilted > 10 degrees. Moreover, deformation of the newly formed spinal canal was observed in 51 (59%) of 86 cases, and bone fusion between the HA spacer and spinous process was achieved in only 15 (8.7%) of 172 cases. Neurological worsening and neck pain, however, were not associated with displacement of HA spacers or deformation of the spinal canal. CONCLUSIONS: In double-door laminoplasty, postoperative displacement of the HA spacer with deformation of the enlarged spinal canal occurred frequently. Hydroxyapatite spacers tend to become displaced after surgery. Placing the HA spacer at the base of the spinous process close to the dura mater may prevent postoperative displacement. PMID- 19558288 TI - Spinal cord oligodendroglioma with 1p and 19q deletions presenting with cerebral oligodendrogliomatosis. AB - Oligodendroglioma of the spinal cord is a rare tumor that most often presents with spinal cord symptoms. The authors present a case of spinal cord oligodendroglioma that was associated with cerebral rather than spinal cord symptoms. A 30-year-old woman developed nausea, vomiting, and severe headaches. Magnetic resonance imaging of the brain showed meningeal enhancement. The patient underwent a craniotomy with biopsies of the meninges and brain. The biopsy findings revealed an abnormal arachnoid thickening without tumor cells. The patient later developed hydrocephalus and underwent shunt placement. Cerebrospinal fluid cytological findings were negative for tumor cells or infection. She was found to have a cervical cord lesion at C3-4 that was initially nonenhancing but later enhanced after Gd administration. Biopsy of the cord lesion with partial resection showed a WHO Grade II oligodendroglioma with 1p and 19q deletions determined by fluorescence in situ hybridization. Neurooncological treatment with tumor radiation and temozolomide (Temodor) resulted in improvement in radiographic findings, symptoms, and long-term survival. This paper presents an extensive review of the literature, which revealed only 2 other reported cases of cerebral symptoms in adults that preceded spinal cord symptoms in a patient with oligodendroglioma of the spinal cord. It is also the first reported case of oligodendrogliomatosis due to a cervical spinal cord oligodendroglioma with 1p and 19q deletions. PMID- 19558289 TI - Financial impact of spinal cord stimulation on the healthcare budget: a comparative analysis of costs in Canada and the United States. AB - OBJECT: Many institutions with spinal cord stimulation (SCS) programs fail to realize that besides the initial implantation cost, budgetary allocation must be made to address annual maintenance costs as well as complications as they arise. Complications remain the major contributing factor to the overall expense of SCS. The authors present a formula that, when applied, provides a realistic representation of the actual costs necessary to implant and maintain SCS systems in Canada and the US. METHODS: The authors performed a retrospective analysis of 197 cases involving SCS (161 implanted and 36 failed trial stimulations) between 1995 and 2006. The cost of patient workup, initial implantation, annual maintenance, and resources necessary to resolve complications were assessed for each case and a unit cost applied. The total cost allocated for each case was determined by summing across healthcare resource headings. Using the same parameters, the unit cost was calculated in both Canadian (CAD) and US dollars (USD) at 2007 prices. RESULTS: The cost of implanting a SCS system in Canada is $21,595 (CAD), in US Medicare $32,882 (USD), and in US Blue Cross Blue Shield (BCBS) $57,896 (USD). The annual maintenance cost of an uncomplicated case in Canada is $3539 (CAD), in US Medicare $5071 (USD), and in BCBS $7277 (USD). The mean cost of a complication was $5191 in Canada (range $136-18,837 [CAD]). In comparison, in the US the figures were $9649 (range $381-28,495) for Medicare and $21,390 (range $573-54,547) for BCBS (both USD). Using these calculations a formula was derived as follows: the annual maintenance cost (a) was added to the average annual cost per complication per patient implanted (b); the sum was then divided by the implantation cost (c); and the result was multiplied by 100 to obtain a percentage (a + b / c x 100). To make this budgetary cap universally applicable, the results from the application of the formula were averaged, resulting in an 18% premium. CONCLUSIONS: For budgeting purposes the institution should first calculate the initial implantation costs that then can be "grossed up" by 18% per annum. This amount of 18% should be in addition to the implantation costs for the individual institution for new patients, as well as for each actively managed patient. This resulting amount will cover the costs associated with annual maintenance and complications for every actively managed patient. As the initial cost of implantation in any country reflects their current economics, the formula provided will be applicable to all implanters and policy makers alike. PMID- 19558290 TI - Spontaneous spinal epidural hematoma inducing acute anterior spinal cord syndrome. AB - Spontaneous spinal epidural hematoma (SSEH) is rare. Its etiology remains controversial; however, spinal venous wall susceptibility to intravenous pressure change and the resultant venous rupture seem to be involved. The authors report a case of SSEH dorsal to the spine producing acute anterior spinal cord syndrome. A posterior SSEH between the C-3 and T-5 levels caused progressive tetraparesis and the disappearance of superficial body sensation below the level of C-8, although deep sensation remained completely intact. This neurological false localizing sign seems to have resulted from counterforce by preexisting asymptomatic cervical intervertebral disc herniation at the C6-7 levels inducing direct pressure on the anterior spinal cord. This case is the first reported instance of posterior cervical SSEH manifesting acute anterior spinal cord syndrome as its false localizing sign. PMID- 19558291 TI - Complications in spinal surgery: comparative survey of spine surgeons and patients who underwent spinal surgery. AB - OBJECT: Definitions of complications in spinal surgery are not clear. Therefore, the authors assessed a group of practicing spine surgeons and, through the surgeons' responses to an online and emailed survey, developed a simple definition of operative complications due to spinal surgery. To validate this assessment, the authors revised their survey to make it appropriate for a lay audience and repeated the assessment with a cohort of patients who underwent spine surgery. METHODS: The authors surveyed a cohort of practicing spine surgeons via email and a web-based survey. Surgeons were presented with various complication scenarios and were asked to grade the presence or absence of a complication as well as complication severity, with responses limited to "major complication" and "minor complication/adverse event." The authors administered a similar assessment, modified for lay persons, to patients in a spinal surgery clinic. RESULTS: Complete responses were obtained from 229 surgeons; orthopedic surgeons comprised the majority of respondents (73%). The authors obtained completed surveys from 197 patients. Overall, there was consistent agreement between physicians and patients regarding the presence or absence of a complication in the majority of scenarios (8 [73%] of 11 scenarios with agreement that a complication was present). The overall kappa value, evaluating major versus minor complication, and presence or absence of a complication over the entire cohort, was fair (kappa = 0.21). The authors found greater variation between the cohorts when evaluating complication severity. Patients were consistently more critical than physicians in the majority of scenarios in which a difference was evident. In 4 scenarios, patients were more likely than surgeons to deem the scenario a complication and to grade the complication as major versus minor (p < 0.01). In 3 additional scenarios, patients were more likely than physicians to grade a major complication as opposed to minor complication (p < 0.01). In only 1 scenario were patients less likely than physicians to report a complication (p < 0.001). CONCLUSIONS: Comparing responses of spine surgeons and patients who underwent spinal surgery in assessing a group of common postoperative events, the authors found significant agreement on perception of presence of a complication in the majority of scenarios reviewed. However, patients were consistently more critical than surgeons when differences in reporting were found. The authors' data underscore the importance of reconciling differing opinions regarding complications through open discussions between physicians and patients to ensure accurate patient expectations of planned medical or surgical interventions. PMID- 19558292 TI - Iatrogenic lumbar pseudoaneurysm causing dural sac compression after spine surgery. AB - A 69-year-old man presented 3 months after lumbar spine surgery with progressive paraparesis and bladder and sphincter dysfunction caused by a lumbar artery pseudoaneurysm compressing the dural sac. After embolization with glue, the thrombosed pseudoaneurysm substantially decreased in size and the patient's neurological symptoms improved. PMID- 19558293 TI - Modified fenestration with restorative spinoplasty for lumbar spinal stenosis. AB - The authors developed an original procedure, modified fenestration with restorative spinoplasty (MFRS) for the treatment of lumbar spinal stenosis. The first step is to cut the spinous process in an L-shape, which is caudally reflected. This procedure allows easy access to the spinal canal, including lateral recesses, and makes it easy to perform a trumpet-style decompression of the nerve roots without violating the facet joints. After the decompression of neural tissues, the spinous process is anatomically restored (spinoplasty). The clinical outcomes at 2 years were evaluated using the Japanese Orthopaedic Association (JOA) scale and patients' satisfaction. Radiological follow-up included radiographs and CT. Between January 2000 and December 2002, 109 patients with neurogenic intermittent claudication with or without mild spondylolisthesis underwent MFRS. Of these, 101 were followed up for at least 2 years (follow-up rate 93%). The average score on the self-administered JOA scale in 89 patients without comorbidity causing gait disturbance improved from 13.3 preoperatively to 22.9 at 2 years' follow-up. Neurogenic intermittent claudication disappeared in all cases. The patients' assessment of treatment satisfaction was "satisfied" in 74 cases, "slightly satisfied" in 12, "slightly dissatisfied" in 2, and "dissatisfied" in 1 case. In 16 cases (18%), a minimum progression of slippage occurred, but no symptomatic instability or recurrent stenosis was observed. Computed tomography showed that the lateral part of the facet joints was well preserved, and the mean residual ratio was 80%. The MFRS technique produces an adequate and safe decompression of the spinal canal, even in patients with narrow and steep facet joints in whom conventional fenestration is technically demanding. PMID- 19558294 TI - Postoperative retroperitoneal hematoma following transforaminal percutaneous endoscopic lumbar discectomy. AB - OBJECT: The purpose of this study was to demonstrate the clinical characteristics of postoperative retroperitoneal hematoma (RPH) following transforaminal percutaneous endoscopic lumbar discectomy (PELD) and to discuss how to prevent the complication of unintended hemorrhage. METHODS: The medical records of 412 consecutive patients treated with transforaminal PELD between January 2005 and May 2007 were reviewed. A total of 4 patients (0.97%) experienced symptomatic postoperative RPH. The clinical outcomes were evaluated using the visual analog scale and the Oswestry Disability Index. RESULTS: The common symptom in all patients with a hematoma was inguinal pain. The mean hematoma volume was 527.9 ml (range 53.3-1274.1 ml). Two patients with massive diffuse-type RPHs compressing the intraabdominal structures required open hematoma evacuation performed by general surgeons, and the other 2 patients with small, localized RPHs of < 100 ml were treated conservatively. The mean follow-up period was 21.3 months (range 13 29 months). The mean visual analog scale score for radicular leg pain improved from 7.6 to 1.8 and that for back pain improved from 4.3 to 2. The mean Oswestry Disability Index improved from 58.8 to 9.1%. The preoperative symptoms improved after the second treatment without significant neurological sequelae in all patients. CONCLUSIONS: Although transforaminal PELD is a minimally invasive and safe procedure, the possibility of RPH should be kept in mind. Adequate technical and anatomical considerations are important to avoid this unusual hemorrhagic complication, especially in the patient with underlying medical problems or previous operative scarring. A high index of suspicion and early detection is also important to avoid the progression of the hematoma. PMID- 19558295 TI - Thoracic and lumbar laminoplasty using a translaminar screw: morphometric study and technique. AB - OBJECT: The aim of this study was to describe a novel technique for laminoplasty in which translaminar screws are used in the thoracic and lumbar spine. METHODS: The authors first performed a morphometric study in 20 control individuals using 3D reconstructed CT scans and spine simulation software to measure the lengths and diameters of the spaces available for translaminar screw placement from the T 1 to S-1. Based on the results of the morphometric study, the authors then attempted translaminar screw fixation in 5 patients (April 2007-July 2007) after en bloc laminectomy in the thoracic and lumbar regions. All patients had intradural lesions: 3 schwannomas, 1 cavernoma, and 1 arachnoid cyst. RESULTS: The morphometric study in control individuals revealed that the safe trajectories for simulated screws measured 25-30 mm in length and 8-11 mm in diameter in the thoracic region (T1-12) and 26-34 mm in length and 6-7 mm in diameter in the lumbosacral region (L1-S1). This morphometric and simulation study showed that translaminar screw placement would be possible in practice. Five patients underwent en bloc laminoplasty and translaminar screw fixation in which the screws measured 2.7 mm in diameter and 24 or 26 mm in length. Sixteen attempts at translaminar fixation were made in 8 vertebrae. Fourteen translaminar screws were successfully placed at the thoracic and lumbar levels. Two microplates had to be used because the laminae were too thin and narrow after further laminectomy with undercutting. There were no complications associated with the translaminar screws. The mean follow-up period was 14.5 months. There was no screw breakage or displacement. Solid osseous fusion was documented in 2 patients who underwent CT scanning 15 months postoperatively. CONCLUSIONS: The authors found that the laminoplasty and translaminar screw technique is feasible in the thoracic and lumbar regions, but further studies are needed to analyze the biomechanical effects and long-term outcomes in a large number of patients. PMID- 19558296 TI - Minimally invasive anterior lumbar interbody fusion followed by percutaneous translaminar facet screw fixation in elderly patients. AB - OBJECT: The purpose of this study was to determine whether anterior lumbar interbody fusion (ALIF) followed by percutaneous translaminar facet screw fixation is effective in elderly patients with degenerative spinal disease. METHODS: Twenty-nine patients > 60 years old who underwent ALIF with percutaneous translaminar facet screw fixation from January to June 2004 were studied. The radiological and clinical data of these patients were collected and analyzed. The mean follow-up period was 14.6 months (range 12-17 months). RESULTS: The mean preoperative, immediate postoperative, and 6- and 12-month postoperative posterior disc heights were 7.1, 11.6, 9.8, and 9.8 mm, respectively. Subsidences of posterior disc height > 20% developed in 9 patients (30%). The significant risk factor for subsidence was found to be 2-level operations (p = 0.023). The mean preoperative Oswestry Disability Index score and visual analog scale scores for the back and leg were 24.4, 6.6, and 7.5, respectively, and improved postoperatively to 14.2, 1.5, and 1.8, respectively. CONCLUSIONS: Minimally invasive ALIF followed by percutaneous translaminar facet screw fixation was performed as a minimally invasive surgical technique in elderly patients. However, in certain circumstances such as multilevel operations or in patients with severe osteoporosis, significant cage subsidence can develop. PMID- 19558297 TI - Prevalence and clinical features of intraspinal facet cysts after decompression surgery for lumbar spinal stenosis. AB - OBJECT: Although many cases of primary intraspinal facet cysts in the lumbar spine have been reported, there have only been a few reports of postoperative intraspinal facet cysts in the lumbar spine. The purpose of this study was to investigate the prevalence and clinical features of postoperative intraspinal facet cysts in the lumbar spine. METHODS: Data from 81 patients undergoing microendoscopic posterior decompression to treat lumbar spinal stenosis were reviewed. The development of a postoperative intraspinal facet cyst was observed using MR imaging during 1 year after surgery. If the patient demonstrated a postoperative intraspinal facet cyst, additional MR imaging was performed to evaluate the natural course of the cyst. Furthermore, the authors conducted a comparative evaluation to identify the factors associated with the causes of cyst development. RESULTS: A postoperative intraspinal facet cyst developed in 7 patients (8.6%) during 1 year after surgery. Spondylotic spinal stenosis, degenerative spondylolisthesis, and degenerative scoliosis were revealed before surgery in 2, 4, and 1 patient, respectively. In 5 patients, the cysts developed within 3 months after surgery. Although 3 patients exhibited symptoms caused by cyst development, all symptoms were relieved by conservative treatment. On radiographic evaluations, postoperative segmental spinal instability, including a progression of spondylolisthesis and disc degeneration, was revealed in 6 (86%) of the 7 patients. Spontaneous regression of the cysts was observed in 5 (71%) of these 7 patients. On comparative evaluation of patients with and without postoperative intraspinal facet cysts, the presence of segmental spinal instability before surgery (including degenerative spondylolisthesis) and the appearance of postoperative segmental spinal instability were related to the development of the cysts. CONCLUSIONS: The prevalence of postoperative intraspinal facet cysts, including asymptomatic cysts, was 8.6% during 1 year after decompression surgery for lumbar spinal stenosis. The development of postoperative intraspinal facet cysts was related to the presence of segmental spinal instability before surgery (including degenerative spondylolisthesis) and postoperative segmental spinal instability, including a progression of spondylolisthesis and disc degeneration after surgery. A postoperative intraspinal facet cyst, which can be expected to regress spontaneously with a probability > 50%, should be recognized as one of the postoperative complications of decompression surgery for lumbar spinal stenosis. PMID- 19558301 TI - Trigeminal neuralgia in the presence of ectatic basilar artery and basilar invagination: treatment by foramen magnum decompression. AB - This 65-year-old woman presented with a 7-year history of classic trigeminal neuralgia. After a conservative therapeutic approach and because the pain had become unbearable, she was subjected to surgical treatment. Examinations revealed an ectatic basilar artery that indented deeply into the region of the root entry zone of cranial nerve V. Additionally, severe basilar invagination, the fusion of multiple cervical vertebrae, and partial agenesis of the clivus were observed. Foramen magnum decompression resulted in lasting relief from the pain. The authors discuss the pathogenesis of trigeminal neuralgia in a relatively rare clinical situation. PMID- 19558298 TI - Biomechanical, biochemical, and histological characterization of canine lumbar facet joint cartilage. AB - OBJECT: Tissue engineering appears to be a promising strategy for articular cartilage regeneration as a treatment for facet joint arthritis. Prior to the commencement of tissue engineering approaches, design criteria must be established to determine the required functional properties of the replacement tissue. As characterization of the functional properties of facet joint cartilage has not been performed previously, the objective of this study was to determine the biomechanical, biochemical, and histological properties of facet joint cartilage. METHODS: The in vitro testing was conducted using 4 lumbar spinal segments obtained from skeletally mature canines. In each specimen, articular cartilage was obtained from the superior surface of the L3-4 and L4-5 facet joints. Creep indentation was used to determine the compressive biomechanical properties, while uniaxial tensile testing yielded the Young modulus and ultimate tensile strength of the tissue. Additionally, biochemical assessments included determinations of cellularity, glycosaminoglycan (GAG) content, and collagen content, as well as enzymelinked immunosorbent assays for collagen I and II production. Finally, histological characterization included H & E staining, as well as staining for collagen and GAG distributions. RESULTS: The means +/- standard deviation values were determined. There were no differences between the 2 spinal levels for any of the assessed properties. Averaged over both levels, the thickness was 0.49 +/- 0.10 mm and the hydration was 74.7 +/- 1.7%. Additionally, the cells/wet weight (WW) ratio was 6.26 +/- 2.66 x 10(4) cells/mg and the cells/dry weight (DW) ratio was 2.51 +/- 1.21 x 10(5) cells/mg. The GAG/WW was 0.038 +/- 0.013 and the GAG/ DW was 0.149 +/- 0.049 mg/mg, while the collagen/WW was 0.168 +/- 0.026 and collagen/DW was 0.681 +/- 0.154 mg/ mg. Finally, the aggregate modulus was 554 +/- 133 kPa, the Young modulus was 10.08 +/- 8.07 MPa, and the ultimate tensile strength was 4.44 +/- 2.40 MPa. CONCLUSIONS: To the best of the authors' knowledge, this study is the first to provide a functional characterization of facet joint articular cartilage, thus providing design criteria for future tissue engineering studies. PMID- 19558302 TI - Wound healing after craniotomy: a randomized trial comparing scalp clips to artery forceps for scalp hemostasis. AB - OBJECT: In this prospective randomized clinical trial, investigators looked at wound healing after craniotomy. The hypothesis was that the self-closing plastic scalp clips used for hemostasis on the skin edge might lead to localized microscopic tissue damage and subsequent delayed wound healing. METHODS: The trial consisted of 2 arms in which different methods were used to secure scalp hemostasis: 1) the routinely used plastic clips (Scalpfix, Aesculap); and 2) the older method of artery forceps placed on the galea. Participants were restricted to those > 16 years of age undergoing craniotomies expected to last > 2 hours. Repeat operations were not included. One hundred fifty patients were enrolled. They were visited at 3 and 6 weeks postoperatively by an observer blinded to the method used, and the wounds were assessed for macroscopic epithelial closure, signs of infection, and hair regrowth by using a predefined assessment scale. RESULTS: The results showed no significant difference in wound healing between the 2 groups at either 3 weeks (OR 0.55, 95% CI 0.27-1.11; p = 0.09) or 6 weeks (OR 0.79, 95% CI 0.39-1.58; p = 0.50). The length of operation was found to be a significant factor affecting wound healing at 6 weeks (OR/hour 0.68, 95% CI 0.51 0.92; p = 0.01). CONCLUSIONS: The use of Aesculap Scalpfix self-retaining plastic scalp clips on the skin edge during craniotomy surgery does not appear to affect wound healing significantly to the postoperative 6-week mark. PMID- 19558303 TI - Gamma knife surgery for lymphocytic hypophysitis. AB - Lymphocytic hypophysitis is a relatively uncommon autoimmune inflammatory disorder affecting the pituitary gland. It most frequently occurs in women of child-bearing age. The authors report on their experience with a patient who presented with diplopia and marked enlargement of the pituitary gland. She underwent transsphenoidal surgery, and histopathological analysis confirmed the diagnosis of lymphocytic hypophysitis. The disease proved refractory to resection, and any attempt at withdrawal of corticosteroid therapy resulted in a return of the patient's symptoms and enlargement of the sellar contents. The patient underwent Gamma Knife surgery (GKS) to the sella and both cavernous sinuses. After GKS, the patient was able to discontinue steroid therapy without return of her symptoms. Follow-up MR images demonstrated no evidence of recurrence of lymphocytic hypophysitis. For persistent lymphocytic hypophysitis, GKS is a reasonable treatment option. PMID- 19558304 TI - Preoperative antiepileptic drug administration and the incidence of postoperative seizures following bur hole-treated chronic subdural hematoma. AB - OBJECT: Despite the prevalence of chronic subdural hematoma (CSDHs) in the rapidly growing elderly population, several aspects of disease management remain unclear. In particular, there is still conflicting evidence regarding the efficacy of antiepileptic drug (AED) prophylaxis in patients with CSDH who undergo bur hole drainage. The authors endeavored to evaluate the efficacy of AED prophylaxis in reducing the incidence of seizures and improving outcome in this patient population. METHODS: A single surgeon's clinical database (E.S.C.) was analyzed for cases involving bur hole drainage for CSDH. Cases involving nonhemorrhagic subdural effusions as well as acute subdural hemorrhages evacuated by craniotomy were excluded from this study. Patient medical records were evaluated for relevant demographic data, medical history, imaging characteristics, clinical details of the treatment, hospital stay, and discharge summaries. RESULTS: The authors included 88 patients with bur hole-treated CSDH. Eleven patients (12.5%) suffered at least 1 seizure between hemorrhage onset and discharge from their treatment hospital admission. Seizures were more frequent in women than men (p = 0.030) and least frequent in patients with right-sided lesions (p = 0.030). In a multiple logistic regression model, preoperative initiation of AED prophylaxis was the only significant predictor of the lower incidence of postoperative seizures (OR 0.10, p = 0.013). However, preoperative initiation of AED prophylaxis did not significantly affect outcome at discharge. CONCLUSIONS: The finding in this study demonstrates that preoperative AED prophylaxis likely reduces the incidence of postoperative seizures in patients with CSDH treated with bur hole drainage. A future prospective randomized study is necessary to evaluate the effect of seizure reduction on clinical outcome. PMID- 19558306 TI - Insulation discontinuity in a vagus nerve stimulator lead: a treatable cause of intolerable stimulation-related symptoms. AB - Discontinuity in the silicone insulation over an electrode of a left vagus nerve stimulator (VNS) allowed the aberrant leak of current to the phrenic nerve and other structures. This resulted in ipsilateral diaphragmatic dysfunction, inability to vocalize, and severe radiating pain into the jaw and upper incisor for the duration of each stimulation. The device was explanted and a new device was implanted. All stimulation-related symptoms ceased immediately. A similar discontinuity in the silicone insulation is the likely explanation for several prior reports of poorly understood pains and phrenic nerve stimulation in patients with VNSs. The findings and analysis of this case establish a rationale for consideration of replacement of the VNS lead in all similarly symptomatic patients. PMID- 19558307 TI - Obesity, gynecological factors, and abnormal mammography follow-up in minority and medically underserved women. AB - BACKGROUND: The relationship between obesity and screening mammography adherence has been examined previously, yet few studies have investigated obesity as a potential mediator of timely follow-up of abnormal (Breast Imaging Reporting and Data System [BIRADS-0]) mammography results in minority and medically underserved patients. METHODS: We conducted a retrospective cohort study of 35 women who did not return for follow-up >6 months from index abnormal mammography and 41 who returned for follow-up < or =6 months in Nashville, Tennessee. Patients with a BIRADS-0 mammography event in 2003-2004 were identified by chart review. Breast cancer risk factors were collected by telephone interview. Multivariate logistic regression was performed on selected factors with return for diagnostic follow up. RESULTS: Obesity and gynecological history were significant predictors of abnormal mammography resolution. A significantly higher frequency of obese women delayed return for mammography resolution compared with nonobese women (64.7% vs. 35.3%). A greater number of hysterectomized women returned for diagnostic follow up compared with their counterparts without a hysterectomy (77.8% vs. 22.2%). Obese patients were more likely to delay follow-up >6 months (adjusted OR 4.09, p = 0.02). Conversely, hysterectomized women were significantly more likely to return for timely mammography follow-up < or =6 months (adjusted OR 7.95, p = 0.007). CONCLUSIONS: Study results suggest that weight status and gynecological history influence patients' decisions to participate in mammography follow-up studies. Strategies are necessary to reduce weight-related barriers to mammography follow-up in the healthcare system including provider training related to mammography screening of obese women. PMID- 19558308 TI - Psychosocial risk factors as contributors to pregnancy-associated death in Virginia, 1999-2001. AB - OBJECTIVE: To determine if substance abuse, mental illness, and domestic violence contributed to preventable pregnancy-associated death and to describe characteristics of women for whom these factors contributed to death. METHODS: The medical records of 121 women who had pregnancy-associated deaths in Virginia between 1999 and 2001 were reviewed. The incidence of substance abuse, mental illness, and domestic violence was noted during systematic review. Multidisciplinary review of cases was conducted to determine if these factors contributed to death and if reasonable changes may have prevented death. RESULTS: The pregnancy-associated maternal mortality ratio for women experiencing substance abuse, mental illness, or domestic violence as contributors to death was 17.1. Thus, for every 100,000 live births in Virginia, 17.1 women had at least one of these factors directly contribute to death. Substance abuse contributed to death in 28.9% of all cases reviewed (pregnancy-associated maternal mortality ratio = 12.2); 45.7% of those deaths were considered preventable. Mental illness contributed to death in 16.5% of cases (pregnancy associated maternal mortality ratio = 6.9), with 50.0% considered preventable. Domestic violence contributed to death in 14.0% of cases (pregnancy-associated maternal mortality ratio = 5.9), with 64.7% of cases considered preventable. Median household incomes and years of education completed varied widely. Pregnancy-associated maternal mortality ratios for each factor were higher among African American women than among white women. CONCLUSIONS: Psychosocial risk assessment with appropriate referral should be completed for all women seeking care regardless of social status, education, or race. PMID- 19558309 TI - Familial risk factors for self-immolation: a case-control study. AB - OBJECTIVE: Women are the primary victims of self-immolation in Iran; between 70 88% of self-immolation patients are women. The aim of this study is to investigate familial risk factors for self-immolation patients. METHODS: In a case-control study, 30 consecutive cases of deliberate self-inflicted burns admitted to the regional Burn Centre (Imam Khomeini hospital in Kermanshah province, Iran) were compared with 30 controls selected from the community and matched by sex, age, and living area. All cases and controls were reviewed for familial variables, including history of suicide in the family, parental divorce, current conflict with parents, parental death, history of mental disorders in parents, history of addiction in parents, parents' marital conflict, marital conflict with spouse, conflict with other members of the family, addictive disorder in spouse, and socioeconomic level of family. RESULTS: Using chi-square test of association, two variables were significantly associated with self immolation. Marital conflict with spouse (OR 7.80, 95% CI 1.84-33.09) and conflict with other members of the family (OR 10.00, 95% CI 2.94-34.00) were associated with increased risk of self-immolation. The associations of other variables in both case and control groups were not statistically significant. CONCLUSIONS: Findings from this study suggest marital conflict with the spouse and conflict with other members of the family are risk factors for self immolation. Screening, identification, and education of at-risk individuals on problem solving and other aspects of coping skills, as well as interpersonal relationships, could be appropriate preventive actions and strategies to reduce self-immolation in Iran. PMID- 19558310 TI - The effects of two metallophthalocyanines on the viability and proliferation of an esophageal cancer cell line. AB - OBJECTIVE: The objective of this study was to establish the influence of two metallophthalocyanine photosensitizers, in their inactive and activated forms, on the cellular reactions of esophageal cancer cells. BACKGROUND DATA: Photodynamic therapy (PDT) is an alternative used in the treatment of cancer. During PDT, the activated compound produces cytotoxic singlet molecular oxygen ((1)O(2)), which ultimately leads to cell death. Esophageal cancer has become one of the most common cancers to occur in the world, and the incidence in South Africa is high, especially within the black male population. METHODS: Optimal photosensitizer concentration was determined by following the viability of esophageal cancer (SNO) cells treated with a range of concentrations of two metallophthalocyanine photosensitizers, GePcSmix and AlPcSmix, activated by irradiation at a fluence of 20 J/cm(2). Changes in cell morphology were observed after treatment with optimal photosensitizer concentrations, and the effect of the treatment on cell proliferation and cytotoxicity were studied. RESULTS: Cell viability decreased in a dose-dependent manner after PDT, while the photosensitizers in their inactive forms did not have an effect on the cells. The altered morphology of cells after PDT was indicative of a necrotic mode of cell death. The optimal photosensitizer concentrations reduced cell proliferation by more than 50% and a significant reduction in cytotoxicity, as detected by lactate dehydrogenase release, was observed following PDT. CONCLUSION: Under the studied parameters PDT using GePcSmix and AlPcSmix in vitro could be a useful therapy for esophageal cancer since the photosensitizers alone caused no damage, but cell death is imminent post-PDT. PMID- 19558311 TI - Head and neck hemangiomas in pediatric patients treated with endolesional 980-nm diode laser. AB - PURPOSE: In pediatric patients with hemangiomas, vascular malformations, or lymphovenous malformations, therapeutic decisions frequently pose a challenge to surgeons, dermatologists, radiologists, and pathologists. We analyze the value of an interdisciplinary approach, and treatment of hemangiomas with an endolesional 980-nm diode laser. PATIENTS AND METHODS: Between 2000 and 2008 we provided interdisciplinary consultation for patients with hemangiomas, lymphangiomas, and vascular anomalies. We treated 250 patients with a 980-nm diode laser. In patients with hemangiomas and venous malformations, clinical diagnosis was confirmed by color Doppler flow imaging and magnetic resonance imaging; angiography was employed for patients with arteriovenous malformations. All patients were either treated by laser surgery or treated conservatively depending on lesion size and behavior. RESULTS: Two hundred fifty pediatric patients were treated; these included 160 patients with hemangiomas, 50 with vascular malformations, and 40 with lymphatic malformation. The treatment results were analyzed by evaluating the decrease in lesion size and the lesion's complete clinical disappearance. All patients had resolution except 38 for whom there was a reduction in lesion size that required a new session. CONCLUSIONS: The interdisciplinary approach increases diagnostic accuracy, and subsequently improves individual treatment for pediatric patients with hemangioma, lymphangioma, and vascular anomalies. The diode laser was found to be useful in the treatment of lesions in pediatric patients in terms of aesthetic results and resolution. PMID- 19558312 TI - Treatment of resistant port-wine stains with a pulsed dual wavelength 595 and 1064 nm laser: a histochemical evaluation of the vessel wall destruction and selectivity. AB - BACKGROUND AND OBJECTIVES: Pulsed dye laser (PDL) is the current treatment of choice for port-wine stains (PWS), but 25-50% of treated lesions do not demonstrate a significant improvement. Hybrid lasers may improve treatment efficacy, especially those using the synergies between PDL and Nd:YAG 1064 nm laser. The objectives of this study were to assess vessel wall damage and epidermal sparing after a dual wavelength treatment with the two lasers, using different laser parameters. MATERIAL AND METHODS: Post-treatment biopsies, after using a laser platform that allows sequential pulses of PDL and Nd:YAG 1064 nm lasers, were performed in five patients with PWS resistant to PDL. The biopsies were stained with nitroblue-tetrazolium chloride (NBTC), using enzymatic activity that stops immediately after cell death and allows a better identification of viable cells. RESULTS: Five patients with PWS and a median age of 33 years were enrolled in this study. Selectivity and efficacy was observed with this dual wavelength approach, with the best results observed with PDL pulses shorter than 10 ms, use of the 10 mm spot, and a second pass with PDL only. CONCLUSIONS: Histochemical studies with NBTC stain can help the laser surgeon establish the best treatment parameters and understand some of the unwanted side effects. The dual wavelength used in this study showed efficacy, but better assessment of treatment parameters, such as the delay between the two lasers, is needed to avoid side effects. PMID- 19558313 TI - Could strontium ranelate have a synergistic role in the treatment of osteoporosis? PMID- 19558315 TI - Fra-1/AP-1 impairs inflammatory responses and chondrogenesis in fracture healing. AB - Inflammation inevitably follows injury of various tissues, including bone. Transgenic overexpression of Fra-1, a component of the transcription factor activator protein-1 (AP-1), in various tissues progressively and globally enhances bone formation, but little is known about the possible effects of Fra 1/AP-1 on fracture healing. We created a transverse fracture of the mouse tibial diaphysis and examined fracture healing radiologically, histologically, and immunologically. Strikingly, fracture union was delayed even though the bone formation rate in callus was higher in Fra-1 transgenic (Tg) mice. In these mice, chondrogenesis around the fracture site was impaired, resulting in accumulation of fibrous tissue, which interferes with the formation of a bony bridge across the callus. Curiously, immediately after fracture, induction of the inflammatory mediators TNF-alpha, interleukin (IL)-6, and Cox-2 was significantly suppressed in Fra-1 Tg mice followed, by the reduced expression of Sox-9 and BMP-2. Because serum prostaglandin E(2) (PGE(2)) levels were dramatically low in these mice, we administered PGE(2) to the fracture site using a slow-release carrier. The accumulation of fibrous tissue in Fra-1 Tg mice was significantly reduced by PGE(2) administration, and chondrogenesis near the fracture site was partially restored. These data suggest that the Fra-1-containing transcription factor AP-1 inhibits fracture-induced endochondral ossification and bony bridge formation presumably through suppression of inflammation-induced chondrogenesis. PMID- 19558316 TI - Osteoclasts and Arthritis. PMID- 19558314 TI - Regulation of osteogenesis-angiogenesis coupling by HIFs and VEGF. AB - Bone is a highly vascularized tissue, but the function of angiogenesis in bone modeling and remodeling is still poorly defined, and the molecular mechanisms that regulate angiogenesis in bone are only partially elucidated. Genetic manipulations in mice have recently highlighted the critical role of the hypoxia inducible-factor/vascular endothelial growth factor pathway in coupling angiogenesis and osteogenesis. In this brief perspective, we review the current understanding of the mechanisms responsible for this coupling. Elucidation of such mechanisms will expand our knowledge of bone development and homeostasis, and it may aid in the design of new therapies for accelerating bone regeneration and repair. PMID- 19558317 TI - Establishment of a cell line derived from a mouse fetal liver that has the characteristic to promote the hepatic maturation of mouse embryonic stem cells by a coculture method. AB - Stromal cells residing in murine fetal livers have the ability to promote the hepatic maturation of murine embryonic stem cells (ESCs) and hepatic progenitor cells (HPCs) 3848 in vitro. These stromal cells were isolated as the CD49f(+/ )CD45(-)Thy1(+)gp38(+) cell fraction. The present study established a murine fetal liver stromal cell line that induced hepatic maturation in mouse ESCs and HPCs. A transgene containing a temperature-sensitive SV40 large T antigen was transfected into the primary fetal liver stromal cells. These immortalized cells, which were named as the gp38-positive and Thy1-positive murine liver stromal (MLSgt) cells, induced both mouse ESCs and HPCs to differentiate into mature hepatocyte-like cells using a coculture method. Since MLSgt is not a cloned cell line, one clone, MLSgt20, was selected as a line with the characteristic to induce hepatic differentiation, which was comparable to its parental stromal cells. The ESC-derived endoderm cells cocultured with the MLSgt20 cells expressed mature hepatocyte-specific gene markers, including glucose-6-phosphatase, tyrosine aminotransferase, tryptophan 2,3-dioxgenase, and cytochrome P450 (CYP1a1, Cyp1b1, Cyp1a2, and Cyp3a11). In addition, these cells also exhibited hepatic functions, such as glycogen storage and ammonia metabolism. Transmission electron microscopy showed that the cocultured ESCs expressed the morphologic features of mature hepatocytes. In conclusion, a cell line was established that has the characteristic to promote the hepatic maturation of mouse ESCs and HPCs by a coculture method. PMID- 19558318 TI - Association between kallikrein-related peptidases (KLKs) and macroscopic indicators of semen analysis: their relation to sperm motility. AB - Human kallikrein-related peptidases (KLKs) are a family of proteases, the majority of which are found in seminal plasma and have been implicated in semen liquefaction. Here, we examined the clinical value of seminal KLKs in the evaluation of semen quality and differential diagnosis and etiology of abnormal liquefaction and/or viscosity. KLK1-3, 5-8, 10, 11, 13, and 14 were analyzed, using highly specific ELISA assays. Samples were categorized into four clinical groups, according to their state of liquefaction and viscosity. Data were compared between the clinical groups and in association with other parameters of sperm quality, including number of motile sperms, straight line speed, sperm concentration, volume, pH, and patient age. Seminal KLKs were found to be differentially expressed in the four clinical groups. Combination of KLK2, 3, 13, and 14 and KLK1, 2, 5, 6, 7, 8, 10, 13, and 14 showed very strong discriminatory potential for semen liquefaction and viscosity, respectively. Liquefaction state was associated with several parameters of sperm motility. Finally, KLK14 was differentially expressed in asthenospermic cases. In conclusion, the expression level of several seminal plasma KLKs correlates with liquefaction and viscosity indicators of semen quality and may aid in their differential diagnosis and etiology. PMID- 19558320 TI - Effect of a quaternary pentamine on RNA stabilization and enzymatic methylation. AB - Extreme thermophiles produce unusually long polyamines, including the linear caldopentamine (Cdp) and the branched pentamine tetrakis(3-aminopropyl)-ammonium (Taa), with the latter containing a central quaternary ammonium moiety. Here we compare the interaction of these two pentamines with RNA by studying the heat denaturation, electrophoretic behavior, and ability of tRNA to be methylated in vitro by purified tRNA methyltransferases under various salt conditions. At concentrations in the micromolar range, branched Taa causes a considerable increase in the melting temperature (T(m)) of yeast tRNA(Phe) transcripts by >20 degrees C, which is significantly greater than stabilization by the linear Cdp. In non-denaturing gel electrophoresis, strong and specific binding to Taa, but not to Cdp, was clearly observed for tRNA(Phe). In both types of experiments, polyamines and monovalent metal ions competed for binding sites. Structural probing revealed no significant conformational changes in tRNA on Taa binding. In post-transcriptional in vitro methylation reactions, the formation of m(2)G/m(2)(2)G by the methyltransferase Trm1p and of m(1)A by TrmIp were not affected or only slightly stimulated by polyamines. In contrast, Taa specifically inhibited Trm4p-dependent formation of m(5)C only in tRNA(Phe), likely by occupying sites that are relevant to RNA recognition by the methyltransferase. PMID- 19558319 TI - A dual role of the N-terminal FQQI motif in GLUT4 trafficking. AB - In adipocytes, the glucose transporter GLUT4 recycles between intracellular storage vesicles and the plasma membrane. GLUT4 is internalized by a clathrin- and dynamin-dependent mechanism, and sorted into an insulin-sensitive storage compartment. Insulin stimulation leads to GLUT4 accumulation on the cell surface. The N-terminal F5QQI motif in GLUT4 has been shown previously to be required for sorting of the protein in the basal state. Here, we show that the FQQI motif is a binding site for the medium chain adaptin micro1, a subunit of the AP-1 adaptor complex that plays a role in post-Golgi/endosomal trafficking events. In order to investigate the role of AP-1 and AP-2 in GLUT4 trafficking, we generated 3T3-L1 adipocytes expressing HA-GLUT4-GFP and knocked down the AP-1 and AP-2 complex by RNAi, respectively. In AP-1 and AP-2 knockdown adipocytes, GLUT4 accumulates at the cell surface in the basal state, consistent with a role of AP-1 in post endosomal sorting of GLUT4 to the insulin-sensitive storage compartment, and of AP-2 in clathrin-mediated endocytosis. Our data demonstrate a dual role of the F5QQI motif and support the conclusion that the AP complexes direct GLUT4 trafficking and endocytosis. PMID- 19558321 TI - The effect of endogenous preproneuropeptide Y leucine 7 to proline 7 polymorphism on growth and apoptosis in primary cultured HUVECs. AB - Neuropeptide Y (NPY) is a universally expressed neuropeptide involved in the regulation of several physiological functions. The rather common leucine7 to proline7 (L7P) polymorphism in the signal peptide of preproNPY is a functional substitution, which changes the processing and release of NPY in cells. The mutation is associated with altered lipid levels and accelerated atherosclerosis in humans. Based on previous studies, we investigated the effect of the Pro7 allele in endothelial cells, which are known to play a role in the development of atherosclerosis. Cell proliferation and apoptosis were studied in primary cultured, genotyped human umbilical vein endothelial cells (HUVECs). Our results indicate that cells with the [p.L7]+[p.P7] genotype seem to have a tendency to be more sensitive to the growth stimulating effect of NPY and less sensitive to the effect of vascular endothelial growth factor compared to cells with the [p.L7]+[p.L7] genotype. Additionally, cells with the [p.L7]+[p.P7] genotype seem to be more sensitive to apoptosis than [p.L7]+[p.L7] cells. We speculate that the L7P substitution in preproNPY might cause a state of cellular pre-senescence, leading to endothelial dysfunction. This might be one reason for the associations of the L7P polymorphism with atherosclerosis and type II diabetes found in clinical studies. PMID- 19558322 TI - Peritumoral administration of GPI-anchored TIMP-1 inhibits colon carcinoma growth in Rag-2 gamma chain-deficient mice. AB - Exogenous application of recombinant TIMP-1 protein modified by addition of a glycosylphosphatidylinositol (GPI) anchor allows efficient insertion of the fusion protein into cell membranes. This 'cell surface engineering' leads to changes in the proteolytic environment. TIMP-1-GPI shows enhanced as well as novel in vitro biological activities including suppression of proliferation, reduced migration, and inhibition of invasion of the colon carcinoma cell line SW480. Treatment of SW480 tumors implanted in Rag (-/-) common gamma chain (-/-) C57BL/6 mice with peritumorally applied TIMP-1-GPI, control rhTIMP-1 protein, or vehicle shows that TIMP-1-GPI leads to a significant reduction in tumor growth. PMID- 19558323 TI - First structural insights into the TpsB/Omp85 superfamily. AB - Proteins of the TpsB/Omp85 superfamily are involved in protein transport across, or assembly into, the outer membrane of Gram-negative bacteria, and their distant eukaryotic relatives exert similar functions in chloroplasts and mitochondria. The X-ray structure of one TpsB transporter, FhaC, provides the bases to decipher the mechanisms of action of these proteins. With two POTRA domains in the periplasm, a transmembrane beta barrel and a large loop harboring a functionally important motif, FhaC epitomizes the conserved features of the super-family. PMID- 19558324 TI - Metzincin's canonical methionine is responsible for the structural integrity of the zinc-binding site. AB - The metzincins constitute a subclan of metalloproteases possessing a HEXXHXXGXXH/D zinc-binding consensus sequence where the three histidines are zinc ligands and the glutamic acid is the catalytic base. A completely conserved methionine is located downstream of this motif. Families of the metzincin clan comprise, besides others, astacins, adamalysins proteases, matrix metallo proteases, and serralysins. The latter are extracellular 50 kDa proteases secreted by Gram-negative bacteria via a type I secretion system. While there is a large body of structural and biochemical information available, the function of the conserved methionine has not been convincingly clarified yet. Here, we present the crystal structures of a number of mutants of the serralysin member protease C with the conserved methionine being replaced by Ile, Ala, and His. Together with our former report on the leucine and cysteine mutants, we demonstrate here that replacement of the methionine side chain results in an increasing distortion of the zinc-binding geometry, especially pronounced in the chi(2) angles of the first and third histidine of the consensus sequence. This is correlated with an increasing loss of proteolytic activity and a sharp increase of flexibility of large segments of the polypeptide chain. PMID- 19558326 TI - Protein targeting by the signal recognition particle. AB - Protein targeting by the signal recognition particle (SRP) is universally conserved and starts with the recognition of a signal sequence in the context of a translating ribosome. SRP54 and FtsY, two multidomain proteins with guanosine triphosphatase (GTPase) activity, are the central elements of the SRP system. They have to coordinate the presence of a signal sequence with the presence of a vacant translocation channel in the membrane. For coordination the two GTPases form a unique, nearly symmetric heterodimeric complex in which the activation of GTP hydrolysis plays a key role for membrane insertion of substrate proteins. Recent results are integrated in an updated perception of the order of events in SRP-mediated protein targeting. PMID- 19558325 TI - Mitochondrial tRNA import--the challenge to understand has just begun. AB - Mitochondrial translation is important for the synthesis of proteins involved in oxidative phosphorylation, which yields the bulk of the ATP made in cells. During evolution most mitochondria-containing organisms have lost tRNA genes from their mitochondrial genomes. Thus, to support the essential process of nuanced mitochondrial translation, mechanisms to actively transport tRNAs from the cytoplasm across the mitochondrial membranes into the mitochondrion have evolved. Here, we review the currently known tRNA import mechanisms, comment on recent discoveries of various import factors, and suggest a rationale for forces that lie behind the evolution of mitochondrial tRNA import. PMID- 19558327 TI - Induced human pluripotent stem cells: promises and open questions. AB - Adult cells have been reprogrammed into induced pluripotent stem (iPS) cells by introducing pluripotency-associated transcription factors. Here, we discuss recent advances and challenges of in vitro reprogramming and future prospects of iPS cells for their use in diagnosis and cell therapy. The generation of patient specific iPS cells for clinical application requires alternative strategies, because genome-integrating viral vectors may cause insertional mutagenesis. Moreover, when suitable iPS cell lines will be available, efficient and selective differentiation protocols are needed to generate transplantable grafts. Finally, we point to the requirement of a regulatory framework necessary for the commercial use of iPS cells. PMID- 19558328 TI - Protein transport across the peroxisomal membrane. AB - The maintenance of peroxisome function depends on the formation of the peroxisomal membrane and the subsequent import of both membrane and matrix proteins. Without exception, peroxisomal matrix proteins are nuclear encoded, synthesized on free ribosomes and subsequently imported post-translationally. In contrast to other translocation systems that transport unfolded polypeptide chains, the peroxisomal import apparatus can facilitate the transport of folded and oligomeric proteins across the peroxisomal membrane. The peroxisomal protein import is mediated by cycling receptors that shuttle between the cytosol and peroxisomal lumen and depends on ATP and ubiquitin. In this brief review, we will summarize our current knowledge on the import of soluble proteins into the peroxisomal matrix. PMID- 19558329 TI - Catalytic properties of recombinant dipeptidyl carboxypeptidase from Escherichia coli: a comparative study with angiotensin I-converting enzyme. AB - Dipeptidyl carboxypeptidase from Escherichia coli (EcDcp) is a zinc metallopeptidase with catalytic properties closely resembling those of angiotensin I-converting enzyme (ACE). However, EcDcp and ACE are classified in different enzyme families (M3 and M2, respectively) due to differences in their primary sequences. We cloned and expressed EcDcp and studied in detail the enzyme's S(3) to S(1)' substrate specificity using positional-scanning synthetic combinatorial (PS-SC) libraries of fluorescence resonance energy transfer (FRET) peptides. These peptides contain ortho-aminobenzoic acid (Abz) and 2,4 dinitrophenyl (Dnp) as donor/acceptor pair. In addition, using FRET substrates developed for ACE [Abz-FRK(Dnp)P-OH, Abz-SDK(Dnp)P-OH and Abz-LFK(Dnp)-OH] as well as natural ACE substrates (angiotensin I, bradykinin, and Ac-SDKP-OH), we show that EcDcp has catalytic properties very similar to human testis ACE. EcDcp inhibition studies were performed with the ACE inhibitors captopril (K(i)=3 nM) and lisinopril (K(i)=4.4 microM) and with two C-domain-selective ACE inhibitors, 5-S-5-benzamido-4-oxo-6-phenylhexanoyl-L-tryptophan (kAW; K(i)=22.0 microM) and lisinopril-Trp (K(i)=0.8 nM). Molecular modeling was used to provide the basis for the differences found in the inhibitors potency. The phylogenetic relationship of EcDcp and related enzymes belonging to the M3 and M2 families was also investigated and the results corroborate the distinct origins of EcDcp and ACE. PMID- 19558330 TI - Epidermal growth factor receptor signaling in liver cell proliferation and apoptosis. AB - Recent evidence suggests that epidermal growth factor receptor (EGFR)-dependent signaling contributes to liver cell proliferation, as well as to apoptotic liver cell death, and represents an important regulator of hepatic regeneration. This article summarizes recent findings on the molecular mechanisms involved in EGFR mediated cell proliferation and apoptosis. The emphasis is on the interplay between EGFR and CD95 (Fas, APO-1) death receptor-signaling, which is determined by the signaling context and liver cell type investigated. PMID- 19558331 TI - Effect of curcumin on amyloidogenic property of molten globule-like intermediate state of 2,5-diketo-D-gluconate reductase A. AB - We identified a molten globule-like intermediate of 2,5-diketo-D-gluconate reductase A (DKGR) at pH 2.5, which has a prominent beta-sheet structure. The molten globule state of the protein shows amyloidogenic property >50 microm protein concentration. Interestingly, a 1:1 molar ratio of curcumin prevents amyloid formation as shown by the Thioflavin-T assay and atomic force microscopy. To the best of our knowledge, this is the first report on amyloid formation by an (alpha/beta)(8)-barrel protein. The results presented here indicate that the molten globule state has an important role in amyloid formation and potential application of curcumin in protein biotechnology as well as therapeutics against amyloid diseases. PMID- 19558332 TI - Mechanism of activation of Saccharomyces cerevisiae calcineurin by Mn2+. AB - Saccharomyces cerevisiae calcineurin (CN) consists of a catalytic subunit CNA1 or CNA2 and a regulatory subunit CNB1. The kinetics of activation of yeast CN holoenzymes and their catalytic domains by Mn2+ were investigated. We report that the in vitro phosphatase reaction activated by Mn2+ typically has a pronounced initial lag phase caused by slow conformational rearrangement of the holoenzyme Mn2+. A similar lag phase was detected using just the catalytic domain of yeast CN, indicating that the slowness of Mn2+-induced conformational change of CN results from a rearrangement within the catalytic domain. The Mn2+-activation of CN was reversible. The dissociation constant of the CN heterodimer containing the CNA2 subunit in the presence of Mn2+ was 3-fold higher than that of CN containing the CNA1 subunit and that of the catalytic domains of CNA1 and CNA2, pointing to differences between the residues surrounding the Mn2+-binding sites of CNA1 and CNA2. PMID- 19558333 TI - Degradable polyethylenimines as DNA and small interfering RNA carriers. AB - Gene therapy is a powerful approach in the treatment of a wide range of both inherited and acquired diseases. Nonviral delivery systems have been proposed as safer alternatives to viral vectors because they avoid the inherent immunogenicity and production problems that are seen when viral systems are used. Many cationic polymers, including high-molecular-weight polyethylenimine (PEI) have been widely studied as gene-delivery carriers, both, in vitro and in vivo. However, interest has recently developed in degradable polymeric systems. The advantage of degradable polymer is its low in-vivo cytotoxicity, which is a result of its easy elimination from the cells and body. Degradable polymer also enhances transfection of DNA or small interfering RNA (siRNA) for efficient gene expression or silencing, respectively. This review paper summarizes and discusses the recent advances with degradable PEIs, such as cross-linked and grafted PEIs for DNA and siRNA delivery. PMID- 19558334 TI - New treatments for chronic obstructive pulmonary disease and viable formulation/device options for inhalation therapy. AB - Chronic obstructive pulmonary disease (COPD) is an increasingly important cause of morbidity and mortality, pathological features of which are pulmonary inflammation and irreversible airflow obstruction. Current therapies for COPD are aimed at improvement of clinical symptoms and reduction of inflammation in the respiratory systems. There is a pressing need for the development of new COPD medication, particularly as no existing treatment has been shown to reduce disease progression. In spite of a better understanding of the underlying disease process, there have been limited advances in the drug therapy of COPD, in contrast to the enormous advances in asthma management. Several new therapeutic targets and strategies have been proposed, and new drug candidates, including bronchodilators, protease inhibitors anti-inflammatory drugs and mediator antagonists, are now in clinical development for COPD treatment. New dry powder inhaler (DPI) systems for inhaled COPD therapy have also been developed to maximize drug concentrations in the airway systems, while minimizing systemic exposure and associated toxicity. This article aims to review recent developments in COPD drugs and the delivery systems for inhalation therapy, with particular emphasis on device options and formulations of DPI systems. PMID- 19558335 TI - RANKL inhibition for the management of patients with benign metabolic bone disorders. AB - The receptor activator of NF-kappaB ligand (RANKL) is a member of the TNF receptor superfamily, essential for osteoclastogenesis. It binds to its receptor activator of NF-kappaB on the surface of osteoclast precursors and enhances their differentiation, survival and fusion, while it activates mature osteoclasts and inhibits their apoptosis. The effects of RANKL are counteracted by osteoprotegerin (OPG), a neutralizing decoy receptor. Derangement of the balance in RANKL/OPG action is implicated in the pathophysiology of metabolic bone diseases, including osteoporosis. Current therapies used to prevent or treat metabolic bone diseases are thought to act, at least in part, through modification of the RANKL/OPG dipole. The idea of using a molecule that could specifically bind and neutralize RANKL to decrease bone resorption and subsequent bone loss is appealing. Recombinant OPG was initially tested. Denosumab, a fully human monoclonal antibody against RANKL, is a promising antiresorptive agent under investigation. It rapidly decreases bone turnover markers resulting in a significant increase in bone mineral density and reduction in fracture risk. However, because receptor activator of NF-kappaB activation by RANKL is also essential for T-cell growth and dendritic-cell function, inhibition of its action could simultaneously affect the immune system, leading to susceptibility in infections or malignancies. PMID- 19558336 TI - Pharmacotherapy of sarcoma. AB - BACKGROUND: Comprising < 1% of adult malignancies and approximately 12% of pediatric malignancies, sarcomas are derived from a variety of connective tissues and exhibit highly variable responsiveness to therapy. The clinical and biologic heterogeneity of the > 50 histologic subtypes of sarcomas often require different therapeutic approaches. OBJECTIVE: This review describes the use of therapeutic agents in the management of bone and soft-tissue sarcomas. METHODS: Relevant literature is identified and presented from major conference proceedings, as well as using the PubMed search engine. RESULTS/CONCLUSIONS: Chemotherapy has improved outcomes over the past few decade, particularly in patients with certain bone sarcomas and gastrointestinal stromal tumors; while in the majority of patients, additional strategies are necessary. PMID- 19558337 TI - Pharmacogenetics in heart failure: promises and challenges. AB - Pharmacotherapy remains the cornerstone in the treatment of heart failure. There is a wide variability in the individual's response to treatment, which is at least partially ascribed to genetic factors. Pharmacogenetics studies the differential clinical effects due to genetic variances. Some effects of the major neurohormonal inhibitors like angiotensin-converting enzyme (ACE)-inhibitors and beta-blockers are importantly modulated by genetic polymorphisms. So far, however, this does not result in standard genetic testing before starting specific therapy. This review discusses several important pharmacogenetic targets. Furthermore, it is argued that new and sophisticated high-throughput genetic screens could be employed to develop pharmacogenetic screening further. Prospective large-scale pharmacogenetic studies are warranted as we believe that they will identify new targets for therapy and specific populations that benefit from specific treatments. PMID- 19558338 TI - The case for pathogen-specific therapy. AB - At the beginning of the twenty-first century, the treatment of microbial diseases is increasingly complicated by drug resistance, the emergence of new pathogenic microbes, the relatively inefficacy of antimicrobial therapy in immunocompromised hosts, and the reemergence of older diseases, often with drug-resistant microbes. Some of these problems can be traced to the switch between pathogen-specific antibacterial therapy and the nonspecific antibacterial therapy that followed the transition from serum therapy to modern antimicrobial chemotherapy. The widespread availability of cheap, effective, nontoxic wide-spectrum antibacterial therapy for almost 75 years fostered a culture of therapeutic empiricism that neglected diagnostic technologies. Despite unquestioned lifesaving efficacy for individuals with microbial diseases, the use of broad-spectrum antimicrobials was associated with fungal superinfections and antibiotic-associated colitis, helped to catalyze the emergence of resistance, and is now tentatively associated in the pathogenesis of certain chronic diseases, including atopy, asthma and - perhaps - certain forms of cancer. This article briefly reviews these trends and suggests that the current strategy of nonspecific therapy is fundamentally unsound because it damages the microflora and - consequently - the human symbiont. The essay argues for the development of immunotherapy and pathogen-specific therapies, especially with regard to bacterial and fungal diseases, and suggests possible routes to that future. PMID- 19558339 TI - Oral antipsychotics for the treatment of schizophrenia: heterogeneity in efficacy and tolerability should drive decision-making. AB - BACKGROUND: Antipsychotic medications are conventionally divided into two groups: First-generation and second-generation antipsychotics (FGAs and SGAs). There is a disagreement about the role of these two groups in the treatment of schizophrenia. OBJECTIVE: To analyze the reasons for the disagreement and to propose a resolution that would improve the treatment of schizophrenia. METHOD: An overview of individual SGAs and several FGAs involved in the current disagreement is presented. Effectiveness studies that contributed to the SGAs versus FGAs disagreement are assessed, and meta-analyses of SGAs and FGAs are reviewed. RESULTS/CONCLUSIONS: Efficacy variations within SGAs and FGAs result in overlaps between the two groups. Regarding safety, FGAs elicit more extrapyramidal side effects and tardive dyskinesia as well as prolactin elevations than SGAs, whereas some SGAs tend to be associated with more weight gain and disturbances in lipid and glucose regulation than FGAs. However, there are again considerable differences between individual agents and overlaps between the two groups in terms of side effects. The classification of antipsychotics into the two groups is no longer useful. The treatment selection for an individual patient should focus on the suitability of an individual antipsychotic for that patient rather than on the group membership of the drug. PMID- 19558340 TI - Pediatric HIV: new opportunities to treat children. AB - BACKGROUND: Treating HIV-infected children remains a challenge due to a lack of treatment options, appropriate drug formulations and, in countries with limited resources, insufficient access to diagnostic tests and treatment. OBJECTIVE: To summarize current data concerning new opportunities to improve the treatment of HIV-infected children. METHODS: This review includes data from the most recently published peer-reviewed publications, guidelines or presentations at international meetings concerning new ways to treat HIV-infected children. RESULTS/CONCLUSIONS: New WHO guidelines recommend starting combination antiretroviral treatment in all infants aged < 1 year. Although this is common practice in some high-income countries, implementation of these recommendations in countries with limited resources is still a challenge. There is still an important gap between the availability of licensed drugs in children compared with adults. There remains a need for further pharmacokinetic studies, and for more pediatric formulations of antiretroviral drugs with improved palatability. PMID- 19558342 TI - Medical prevention of stroke and stroke recurrence in patients with TIA and minor stroke. AB - BACKGROUND: Secondary stroke prevention after transient ischemic stroke (TIA) or minor stroke is of major importance in order to avoid recurrent cerebrovascular events and decrease morbidity and mortality. OBJECTIVE/METHODS: Systematically review of recently published, high-quality studies emphasizing the need for emergency assessment and treatment of patients with TIA and minor stroke and to give a comprehensive and distinct overview over medical secondary stroke prevention trials performed in these patients. RESULTS/CONCLUSIONS: Evaluation and implementation of preventive stroke therapy has to be immediate in patients with TIA and stroke. For patients with non-cardioembolic stroke, antiplatelet agents are the treatment of choice. Aspirin plus extended-release dipyridamole and clopidogrel are more effective than aspirin and should be used in patients with a high risk of recurrent stroke. Oral anticoagulation is highly effective in patients with a cardiac source of embolism. Treatment of risk factors such as arterial hypertension and high cholesterol is even more important in secondary stroke prevention than in primary prevention. Vitamin supplementation and lowering of elevated levels of homocysteine are not effective in stroke prevention. PMID- 19558341 TI - Emedastine difumarate: a review of its potential ameliorating effect for tissue remodeling in allergic diseases. AB - BACKGROUND: Emedastine difumarate, a selective histamine-H1 receptor antagonist and effective antiallergic agent, inhibits various clinical symptoms of allergic rhinitis, allergic conjunctivitis, urticaria, allergic dermatitis, pruritus cutaneous, and prurigo. In addition to greater efficacy than other antihistamines, emedastine difumarate produces no adverse cardiovascular effects and exhibits minimal anticholinergic activity. Moreover, a recent study revealed that the effect of emedastine difumarate on inhibition of histamine-induced collagen synthesis in vitro was greater in dermal fibroblasts than in nasal mucosa fibroblasts. This result indicates that there are tissue-specific effects of emedastine difumarate and that it may be more effective for treating fibrosis in skin than in nasal mucosa. However, the mechanism and role of tissue remodeling is less well established for allergic skin diseases and allergic conjunctivitis, in comparison to respiratory allergic diseases. OBJECTIVE: This review outlines the involvement of histamine in the pathogenesis of tissue remodeling in a variety of organs, and presents the evidence for the effect of antihistamines on this process. Furthermore, this review also discusses antihistamines as an intervention strategy in tissue remodeling. METHODS: The scientific literature, published abstracts, and selected textbooks were reviewed. RESULTS/CONCLUSION: Although additional evidence is required, emerging evidence suggests that emedastine difumarate may be of value in the prevention of excess tissue remodeling in allergic skin inflammation. PMID- 19558343 TI - Can the bowel substitute for the kidney in advanced renal failure? AB - This Editorial provides a brief commentary exploring the possibilities of the bowel acting as a substitute for kidney function in chronic kidney disease/end stage renal disease. Three concepts are highlighted as potential means by which bowel function can serve as an alternative to renal filtration: oral sorbents, diarrhea therapy and bacterial-enzyme nitrogen recycling within the gut. PMID- 19558344 TI - Probiotic dietary supplementation in patients with stage 3 and 4 chronic kidney disease: a 6-month pilot scale trial in Canada. AB - AIM: This was a pilot clinical trial to assess biochemical and clinical effects of an oral probiotic dietary supplement in chronic kidney disease (CKD) patients (stages 3 and 4). METHODS: A prospective, randomized, double-blind, crossover, placebo-controlled, 6-month trial of probiotic bacteria was conducted in 16 outpatients in Ontario, Canada. Primary endpoints included effect on hematologic, biochemical, and fecal variables, and on general well-being as assessed by quality of life (QOL). These outcomes were evaluated from biochemical parameters, mainly blood urea nitrogen (BUN), creatinine, uric acid, and C-reactive protein (CRP) as a general inflammatory marker. QOL was assessed on a subjective scale of 1 to 10 as the secondary parameter. TRIAL REGISTRATION: This pilot study forms part of registered trial NCT00760162. RESULTS: A total of 13 patients completed the study. Three patients dropped out: one was the receiver of a transplant. The second dropped out for unknown reasons and the third died of myocardial infarction (unrelated to probiotic bacteria or the protocol). Among the 13 patients who completed the trial, the mean change in BUN concentration during the probiotic treatment period (-2.93 mmol/L) differed significantly (p = 0.002) from the mean change in BUN concentration during the placebo period (4.52 mmol/L). In addition, the mean changes in uric acid concentration were moderate during the KB period (24.70 micromol/L) versus during the placebo period (50.62 micromol/L, p = 0.050), and the changes in serum creatinine concentration were insignificant. Neither gastrointestinal nor infectious complications were noted in any subject with improved QOL. CONCLUSION: Orally administered probiotic bacteria selected to metabolize nitrogenous wastes may be tolerated for as long as 6 months. A major limitation of this trial is its small size that may have precluded detection of changes in other biochemical or hematologic parameters that would be evident in larger cohorts. Extension of the evaluation of this probiotic bacterial mixture will include a dose escalation trial in a similar prospective, placebo controlled, and double-blind study site. PMID- 19558346 TI - Synthesis of a novel series of benzylether-containing cinnamoyl derivatives as histone deacetylase inhibitors. AB - Histone deacetylases (HDACs) play an important role in gene transcription, cellular proliferation, apoptosis, and tumorigenesis. A novel series of benzylether-containing cinnamoyl derivatives were designed and synthesized as inhibitors of histone deacetylases (HDACs). Most of these compounds possessed inhibitory activity against the enzymes with IC50 values as low as 30 microM. In addition, compounds 4d (IC50 = 11.1 microM) and 4n (IC50 = 7.7 microM) exhibited high antiproliferative activity against tumor cell growth and effectively induced cell cycle arrest. PMID- 19558347 TI - Stability evaluation of temoporfin-loaded liposomal gels for topical application. AB - Temoporfin (mTHPC) is a potent second-generation synthetic photosensitizer. Topical delivery of mTHPC is of great interest for the photodynamic therapy of psoriasis and superficial skin cancer lesions. The aim of this study was to evaluate the stability of hydrophilic gels containing mTHPC-loaded liposomes. Two different mTHPC-loaded liposome dispersions, composed of 15 % (w/w) nonhydrogenated soybean lecithin of different phosphatidylcholine content, were prepared and incorporated (2:1 w/w) into hydrogels of different carbomer concentrations (1.5, 2.25, and 3%; w/w). Obtained liposomal hydrogels, containing 0.15% (w/w) mTHPC, 10% (w/w) phospholipids, and 0, 0.5, or 1% (w/w) carbomer, were analyzed for flow properties, liposome particle size, and polydispersity index (PDI), pH value, and mTHPC content after their preparation and at predetermined time intervals during 6 months of storage at 4 and 23 degrees C. All hydrogels showed, during the whole period of investigation, adequate characteristics for topical application (i.e., they revealed shear-thinning plastic flow behavior). Rheological parameters, particle size, and PDI of liposomes in hydrogels, mTHPC content, and pH value did not show remarkable changes during the storage of gels, which could make them unacceptable for topical use. The obtained results indicated physical and chemical stability of liposomal gels containing mTHPC during 6 months of storage at both temperatures. PMID- 19558356 TI - Evaluation of poly(D,L-lactide-co-glycolide) microspheres for the lung-targeting of yuanhuacine, a novel DNA topoisomerase I inhibitor. AB - The present study was intended to develop poly(D,L-lactide-co-glycolide) (PLGA; 50:50, 0.15 dL/g) microspheres (MS) loaded with yuanhuacine (YHC) for passive targeting in lung as well as providing a simple evaluation method for the targeting efficiency of MS. A kind of photochromic spiropyran dye was applied to label MS to clearly demonstrate the in vivo distribution characteristics through intravenous injection into mice and rabbits. Sections of 10-microm thickness from different organs were cut using a microtome, and fluorescent microscopy was used to determine the biodistribution of the MS. The average particle size of MS was 9.0 microm, and the glass transition temperature was 37-40 degrees C. In vitro, the cumulative release achieved 50.8% in 24 h. Histological sections from different organs indicated that the amount of MS in lung achieved maximum in 6 h, as about 8 times as in liver and 70 times higher than the average concentration of other organs. In vivo, MS were gradually swelled and drug concentration remained just 10% in 12 h, which would not result in long time embolization in the lung. This evaluation method supplies a simple and visualized channel in focus for the targeting efficiency of PLGA MS. PMID- 19558357 TI - Synthesis, in vitro and in vivo characterization of glycosyl derivatives of ibuprofen as novel prodrugs for brain drug delivery. AB - New glycosyl derivatives of ibuprofen (I, II, III, and IV) were synthesized in order to overcome the ineffective delivery of ibuprofen across the blood-brain barrier owing to its low permeability, using d-glucose as a drug targeting agent. Ibuprofen was linked directly to the C-2, C-3, C-4, and C-6 positions of glucose via ester bonds. Furthermore, in vitro stabilities of the four ester derivatives were evaluated to determine both their stability in aqueous medium and their feasibility to undergo enzymatic cleavage by esterase in biological samples to regenerate the original drug. From the obtained results, compounds I-IV appeared to be moderately stable in pH 7.43 buffer solution, rat plasma, and brain tissue extracts. In vivo experiments showed that the AUC(0-)(t) of ibuprofen in plasma after the injection of prodrugs is several times higher than that of AUC(0-)(t) after the injection of ibuprofen. In addition, the maximal concentration of ibuprofen in brain after the administration of ester IV was three fold higher than that of the control group. Also, the concentration of ibuprofen was kept stable in brain for about 4 h for four esters, which was beneficial for the treatment of Alzheimer's disease and highlighted the possibility of brain drug delivery of ibuprofen using prodrug strategies. PMID- 19558358 TI - A novel recombinant protein TAT-GFP-KDEL with dual-function of penetrating cell membrane and locating at endoplasm reticulum. AB - Although the potential value of phenotypic/functional knockout technology with intrabody/kine in prevention and cure of some serious diseases, such as AIDS and cancer, is being regarded, there are still several technical difficulties. One of the the most critical problems is how to directly deliver the intrabody/kine proteins into endoplasm reticulum (ER). In this study, a novel recombinant protein, TAT-GFP-KDEL, was designed and constructed. In this recombinant protein, HIV-derived TAT (47-57) and an ER retention four-peptide sequence KDEL were fused at the N-terminal and C-terminal of GFP respectively. The results showed that TAT GFP-KDEL had been successfully expressed in bacteria BL21 and its purity reached to 95%. Moreover, we observed that this recombinant protein was able to efficiently transduce into MOLT-4 cells and accurately locate at ER. This study may provide an available strategy to promote the transmembrane delivery and ER localization of protein-based intrabody/kine. PMID- 19558359 TI - Polybutylcyanoacrylate magnetic nanoparticles as carriers of adriamycin. AB - OBJECTIVE: The present study was designed to prepare and evaluate adriamycin polybutylcyanoacrylate magnetic nanoparticles (ADR-PBCA-MNPs) as novel carriers of adriamycin. METHODS: ADR-PBCA-MNPs was prepared by the emulsion polymerization technique. Entrapment efficiency (ER) and drug load (DL) of nanoparticles, along with in vitro release were studied. Pharmacokinetic analysis was carried out in Kunming mice, with blood obtained at determined time points post administration. Biodistribution and recovery rate of ADR was measured and determined. RESULTS: Nanoparticles were visible as approximate spherical particles with good disparity, with an average diameter of 184.6 nm, a minimum diameter of 59.07 nm, and a maximum diameter of 291.66 nm as demonstrated by transmission electron microscopy. The ER and quantity of DL of ADR-PBCA-MNPs were 90.73 and 10.68%, respectively, measured by an ultraviolet-visible light spectrophotometer. In vitro study demonstrated that the release reached a balance after 72 h, with a total release rate of approximately 80%. As shown in pharmacokinetic studies in rats,the ADR-PBCA-MNPs group displayed a slowed doxorubicin release associated with better bioavailability. ADR-PBCA-MNPs reduced ADR accumulation at nontarget sites in the magnetic field, contributing to the reduced toxicity and side effects of ADR. CONCLUSION: ADR-PBCA-MNPs was successfully prepared and had a satisfactory targeted effect under the magnetic field, which can increase ADR concentration at target sites but not at non-target sites. As a result, the therapeutic effect of ADR may be greatly enhanced with minimized drug toxicity and side effects. PMID- 19558360 TI - Encapsulation of adipogenic factors to promote differentiation of adipose-derived stem cells. AB - Insulin and dexamethasone were encapsulated in poly(lactic-co-glycolic acid) (PLGA) microspheres to induce adipogenesis for potential applications in soft tissue reconstruction. Release kinetics and bioactivity of the drugs were examined. Surface morphology and diameter of the PLGA microspheres was evaluated using scanning electron microscopy. The release of insulin was determined using ELISA whereas the release of dexamethasone was evaluated spectrophotometrically. The activity of the drugs was assessed by releasing the drugs in the presence of human adipose-derived stem cells. The ability of the cells cultured with microspheres to differentiate into adipocytes was evaluated using Oil Red O stains. Cells treated with the dexamethasone and insulin microspheres demonstrated a significant increase in lipid inclusions compared with control groups. Insulin and dexamethasone microspheres can reproduce the adipogenic effect exerted by differentiation medium, and may represent a clinically relevant method of stimulating adipogenesis in tissue engineering therapies. PMID- 19558361 TI - Augmented inhibitory effect of superoxide dismutase on superoxide anion release from macrophages by chemical modification with polysaccharide and attenuation effects on radiation-induced inflammatory cytokine expression in vitro. AB - To improve the ability of superoxide dismutase (SOD) to suppress reactive oxygen species (ROS)-mediated injury, chemically modified derivatives of SOD with N,N,N trimethyl chitosan chloride (TMC) and heparin, cationized SOD (TMC-SOD), and anionized SOD (heparin-SOD) were designed and prepared. In this study, the inhibitory effect of TMC-SOD and heparin-SOD on superoxide anion release from macrophages was studied in vitro. Both TMC-SOD and heparin-SOD exhibited excellent inhibitory effects on superoxide anion release from macrophages, and the effects of TMC-SOD surpassed those of native SOD and heparin-SOD. The effects of TMC-SOD and heparin-SOD on inflammatory cytokine expression in vitro were also evaluated. The results showed that both TMC-SOD and heparin-SOD could significantly lower the levels of transforming growth factor-beta1 (TGF-beta1) and interleukine-1beta (IL-1beta) expressed by irradiated 3T3 fibroblasts. These results demonstrated that cationic polysaccharide or anionic polysaccharide SOD derivatives might be useful in the prevention and treatment of ROS-mediated inflammatory diseases. This study also demonstrated that chemical modification of SOD, especially cationization, greatly enhanced SOD's intracellular delivery and, consequently, produced a significant protective effect against ROS-mediated injury. PMID- 19558362 TI - Optimized prodrug approach: a means for achieving enhanced anti-inflammatory potential in experimentally induced colitis. AB - In the present study, prodrug of ketoprofen was synthesized and evaluated in vitro to optimize the prodrug, and in vivo to observe the reduction in gastrointestinal disturbance and enhanced colonic anti- inflammatory potential for the prodrug. The prodrug was synthesized by coupling ketoprofen with L glycine (KET-GLY). In vitro reversion of KET-GLY to ketoprofen was carried out in different pHs and in pH 6.8 containing optimized rat fecal material. In vivo healing potential of KET-GLY was evaluated in acetic acid-induced experimental colitis model. In vitro reversion studies suggested that KET-GLY remained intact in stomach but released the free drug at pH 6.8 containing fresh rat fecal material, where the colonic microfloral enzymes (amidase) hydrolyzed the KET-GLY amide linkage, releasing the free drug. In vivo evaluation indicated KET-GLY to be less toxic in stomach, with enhanced anti-inflammatory potential in the colonic region. These findings suggested KET-GLY to be better in action compared with the parent drug. PMID- 19558363 TI - Enhanced protection of Ins-1 beta cells from apoptosis under hypoxia by delivery of DNA encoding secretion signal peptide-linked exendin-4. AB - In this study, we developed an expression system of exendin-4, a glucagon-like peptide (GLP-1) analog, using a secretion signal peptide (SP) to facilitate exendin-4 secretion. For delivery of the exendin-4 expression system, high molecular-weight polyethylenimine (25 kDa, PEI25k), low-molecular-weight polyethylenimine (2 kDa, PEI2k), and polyamidoamine (PAMAM) dendrimers were evaluated as gene carriers to Ins-1 beta cells. As a result, PEI25k showed the highest transfection efficiency. For the construction of the exendin-4 expression vector, DNA coding the SP sequence was inserted upstream of the exendin-4 cDNA, resulting in the construction of pbeta-SP-Ex-4. Transfection assay showed that the secretion level of exendin-4 increased in the pbeta-SP-Ex-4 transfected cells, compared with the pbeta-Ex-4 transfected cells. To identify the beta-cell protection effect of pbeta-SP-Ex-4 delivery, the Ins-1 beta cells were transfected with pbeta-SP-Ex-4 or pbeta-Ex-4 and incubated under normoxia or hypoxia. An MTT assay showed that the pbeta-SP-Ex-4 transfected cells had higher beta-cell viability than the pbeta-Ex-4 transfected cells under hypoxia. In addition, the pbeta-SP-Ex-4 transfected cells exhibited lower caspase-3 activity than the pbeta-Ex-4 transfected cells. Therefore, PEI25k/pbeta-SP-Ex-4 complex may be useful to protect isolated beta cells from apoptosis during transplantation. PMID- 19558367 TI - Human Ccr4-Not complexes contain variable deadenylase subunits. AB - The Ccr4-Not complex is evolutionarily conserved and important for regulation of mRNA synthesis and decay. The composition of the yeast complex has been well described. Orthologues of the yeast Ccr4-Not components have been identified in human cells including multiple subunits with mRNA deadenylase activity. In the present study, we examine the composition of the human Ccr4-Not complex in an in depth proteomic approach using stable cell lines expressing tagged CNOT proteins. We find at least four different variants of the human complex, consisting of seven stable core proteins and mutually exclusive associated mRNA deadenylase subunits. Interestingly, human CNOT4 is in a separate approximately 200 kDa complex. Furthermore, analyses of associated proteins indicate involvement of Ccr4-Not complexes in splicing, transport and localization of RNA molecules. Taken together, human Ccr4-Not complexes are heterogeneous in composition owing to differences in their deadenylase subunits, which may reflect the multi functionality of these complexes in cellular processes. PMID- 19558366 TI - Origin of pyrimidine deoxyribonucleotide pools in perfused rat heart: implications for 3'-azido-3'-deoxythymidine-dependent cardiotoxicity. AB - In adult non-replicating tissues such as heart, demand for dNTPs (deoxynucleoside triphosphates) is low but essential for mitochondrial DNA replication and nuclear DNA repair. dNTPs may be synthesized from salvage of deoxyribonucleosides or by reduction of ribonucleotides. We have hypothesized that the cardiac mitochondrial toxicity of the nucleoside analogue AZT (3'-azido-3'-deoxythymidine; known as zidovudine) is caused by inhibition of thymidine kinase 2 of the salvage pathway and subsequent TTP pool depletion. The extent to which this hypothesis has merit depends on how much the heart relies on thymidine phosphorylation for maintenance of the TTP pool. In the present study, we used isotopic tracing to demonstrate that both TTP and dCTP are solely synthesized by phosphorylation of thymidine and deoxycytidine respectively, with no evidence for synthesis from other precursors. We have also shown that UTP and CTP are synthesized by phosphorylation of uridine and cytidine respectively, with no detectable role for the de novo pyrimidine synthesis pathway. Lastly, we have demonstrated that AZT decreased the TTP pool by 50% in 30 min of perfusion, while having no effect on other dNTPs. In summary, the present study demonstrated that adult rat heart has a limited mechanism for dCTP and TTP synthesis and thus these pools may be more sensitive than replicating cells to drugs such as AZT that affect the salvage pathway. PMID- 19558368 TI - DNA damage signalling recruits RREB-1 to the p53 tumour suppressor promoter. AB - Transcriptional regulation of the p53 tumour suppressor gene plays an important role in the control of the expression of various target genes involved in the DNA damage response. However, the molecular basis of this regulation remains obscure. In the present study we demonstrate that RREB-1 (Ras-responsive-element-binding protein-1) efficiently binds to the p53 promoter via the p53 core promoter element and transactivates p53 expression. Silencing of RREB-1 significantly reduces p53 expression at both the mRNA and the protein levels. Notably, disruption of RREB-1-mediated p53 transcription suppresses the expression of the p53 target genes. We also show that, upon exposure to genotoxic stress, RREB-1 controls apoptosis in a p53-dependent manner. These findings provide evidence that RREB-1 participates in modulating p53 transcription in response to DNA damage. PMID- 19558369 TI - Multivariate analysis of factors associated with post-anesthetic times to standing in isoflurane-anesthetized horses: 381 cases. AB - OBJECTIVE: To identify anesthesia-related variables which may independently predict time to standing in horses anesthetized with ketamine/diazepam/isoflurane. STUDY DESIGN: Retrospective case series. ANIMALS: Three hundred and eighty-one horses. METHODS: Case records were searched for the years 2000-2003 and 381 horses older than 12 months which weighed at least 200 kg were identified. Data were extracted from the records, and only horses that were anesthetized with xylazine, ketamine, diazepam and isoflurane were included in the analysis. Multiple linear regression was used to relate time to standing with demographic, intraoperative and anesthetic variables. RESULTS: Most (326; 86%) horses recovered unassisted and 55 (14%) were assisted in recovery. The model for unassisted recovery had an R(2) of 0.228 with colic (p < 0.0001), anesthesia duration (p < 0.02), temperature nadir (p < 0.02) and duration of hypotension (p < 0.0001) being significant predictors of time to standing. The final model for predicting assisted recovery time had an R(2) of 0.314 with emergency status (p < 0.0001), warm-blood breed (p < 0.04) and intraoperative administration of ketamine (p < 0.004) being the significant predictor. CONCLUSIONS AND CLINICAL RELEVANCE: Variables which could be impacted by the anesthetist which would result in a faster time to standing include duration of anesthesia, hypothermia and intraoperative hypotension. However, the contribution of anesthesia factors explained <23% of the variability in recovery time, suggesting that other, more important factors contribute to anesthesia recovery time in horses. PMID- 19558370 TI - Effect of lidocaine on the minimum alveolar concentration of sevoflurane in dogs. AB - OBJECTIVE: To investigate the effects of a low-dose constant rate infusion (LCRI; 50 microg kg(-1) minute(-1)) and high-dose CRI (HCRI; 200 microg kg(-1) minute( 1)) lidocaine on arterial blood pressure and on the minimum alveolar concentration (MAC) of sevoflurane (Sevo), in dogs. STUDY DESIGN: Prospective, randomized experimental design. ANIMALS: Eight healthy adult spayed female dogs, weighing 16.0 +/- 2.1 kg. METHODS: Each dog was anesthetized with sevoflurane in oxygen and mechanically ventilated, on three separate occasions 7 days apart. Following a 40-minute equilibration period, a 0.1-mL kg(-1) saline loading dose or lidocaine (2 mg kg(-1) intravenously) was administered over 3 minutes, followed by saline CRI or lidocaine LCRI or HCRI. The sevoflurane MAC was determined using a tail clamp. Heart rate (HR), blood pressure and plasma concentration of lidocaine were measured. All values are expressed as mean +/- SD. RESULTS: The MAC of Sevo was 2.30 +/- 0.19%. The LCRI reduced MAC by 15% to 1.95 +/- 0.23% and HCRI by 37% to 1.45 +/- 0.21%. Diastolic and mean pressure increased with HCRI. Lidocaine plasma concentration was 0.84 +/- 0.18 for LCRI and 1.89 +/- 0.37 microg mL(-1) for HCRI. Seventy-five percent of HCRI dogs vomited during recovery. CONCLUSION AND CLINICAL RELEVANCE: Lidocaine infusions dose dependently decreased the MAC of Sevo, did not induce clinically significant changes in HR or arterial blood pressure, but vomiting was common during recovery in HCRI. PMID- 19558371 TI - Psychometric evaluation of the Diabetes Symptom Checklist-Revised (DSC-R)--a measure of symptom distress. AB - OBJECTIVE: To assess the psychometric validity, reliability, responsiveness, and minimal important differences of the Diabetes Symptoms Checklist-Revised (DSC-R), a widely used patient-reported outcome measure of diabetes symptom distress. RESEARCH DESIGN AND METHODS: Psychometric validity of the DSC-R was assessed using blinded data from a large-scale trial of approximately 4000 type 2 diabetes patients. Confirmatory factorial analysis (CFA) and multitrait analysis were used to examine the construct validity of the structure of DSC-R. DSC-R internal consistency, discriminative validity, and responsiveness were also assessed. Distribution and anchor-based methods were used to estimate minimal important differences for DSC-R domains. RESULTS: Mean age of the sample was 56 years, 42% were female, 88% were Caucasian. Patients had a mean body mass index (BMI) of 32.2 and mean glucose-fasting level of 151.7 md/dl. CFA and multitrait analysis indicated that the scoring of the DSC-R has acceptable construct validity. Item scale correlations ranged from 0.44 to 0.78. Cronbach's alpha coefficients ranged from 0.69 to 0.87. At baseline, DSC-R scores were higher among patients with higher BMI scores (P < 0.0001), supporting the discriminative validity of the DSC R. Minimal important difference estimates ranged from 0.39 to 0.60 points when using distribution methods and from 0.00 to 0.33 when estimated using anchor based methods. CONCLUSIONS: The DSC-R demonstrated excellent psychometric properties when tested in a large-scale diabetes clinical trial. Responsiveness and test-retest reliability of the DSC-R warrant further evaluation. PMID- 19558372 TI - Mapping the cancer-specific EORTC QLQ-C30 to the preference-based EQ-5D, SF-6D, and 15D instruments. AB - OBJECTIVES: To estimate models, via ordinary least squares regression, for predicting Euro Qol 5D (EQ-5D), Short Form 6D (SF-6D), and 15D utilities from scale scores of the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30). METHODS: Forty-eight gastric cancer patients, split up into equal subgroups by age, sex, and chemotherapy scheme, were interviewed, and the survey included the QLQ-C30, SF-36, EQ-5D, and 15D instruments, along with sociodemographic and clinical data. Model predictive ability and explanatory power were assessed by root mean square error (RMSE) and adjusted R(2) values, respectively. Pearson's r between predicted and reported utility indices was compared. Three random subsamples, half in size the initial sample, were created and used for "external" validation of the modeling equations. RESULTS: Explanatory power was high, with adjusted R(2) reaching 0.909, 0.833, and 0.611 for 15D, SF-6D, and EQ-5D, respectively. After normalization of RMSE to the range of possible values, the prediction errors were 12.0, 5.4, and 5.6% for EQ-5D, SF-6D, and 15D, respectively. The estimation equations produced a range of utility scores similar to those achievable by the standard scoring algorithms. Predicted and reported indices from the validation samples were comparable thus confirming the previous results. CONCLUSIONS: Evidence on the ability of QLQ-C30 scale scores to validly predict 15D and SF-6D utilities, and to a lesser extent, EQ-5D, has been provided. The modeling equations must be tried in future studies with larger and more diverse samples to confirm their appropriateness for estimating quality-adjusted life-year in cancer patient trials including only the QLQ-C30. PMID- 19558373 TI - The cost-effectiveness of bosentan in the United Kingdom for patients with pulmonary arterial hypertension of WHO functional class III. AB - OBJECTIVES: To assess whether bosentan or no active intervention, in addition to palliative care, is the more cost-effective first-line treatment option for patients with idiopathic pulmonary arterial hypertension (iPAH) or PAH associated with connective tissue disease (PAH-CTD) of WHO functional classification (FC) III in the United Kingdom. METHODS: A cost-utility model simulated the treatment of patients with PAH of FC III. Patients remained on the selected intervention until death or clinical deterioration to FC IV, which would trigger initiation of epoprostenol treatment. The initial first-line treatment choice was assumed to not affect survival, but to affect the time until clinical deterioration, with this assumption being relaxed in sensitivity analyses. The distribution of time to clinical deterioration was estimated from long-term clinical trial databases of bosentan and from published literature. Utility associated with FC was taken from published literature. Costs were sourced from published literature and from specialist PAH centers. The time horizon was that of patients' lifetimes, with costs and benefits discounted at 3.5% per annum. RESULTS: In the base case, bosentan dominated no active intervention because of the longer time to clinical deterioration and therefore the reduced time, per patient, spent in FC IV, which was associated with high costs of epoprostenol and reduced utility. In sensitivity analyses, bosentan was estimated to be more cost-effective than no active intervention, provided that any survival benefit was not greater than 2 years for patients with iPAH and 1 year for those with PAH-CTD. CONCLUSIONS: Bosentan is likely to be a more cost-effective first-line therapy for patients with PAH FC III in the UK than no active intervention. PMID- 19558374 TI - Intermittent preventive treatment of malaria in infants: how does it work and where will it work? AB - We discuss the potential public health impact of IPTi by estimating the cases of malaria, anaemia and hospital admissions likely to be averted in different transmission settings; and we review the mechanism of action, choice of drugs regimens, and the effect on immunity of IPTi. IPTi using an efficacious drug is likely to substantially reduce cases of clinical malaria in moderate to high transmission settings. However, geographical heterogeneity in malaria transmission could hamper rolling out IPTi as a national policy. PMID- 19558375 TI - Case of fulminant leptospirosis in a renal transplant patient. AB - We present a case of fulminant leptospirosis that was acquired in the suburban area by a 48-year-old male renal transplant recipient. He developed acute renal and hepatic failure with profound jaundice. Spirochetes were identified on liver biopsy. Weil's disease was suspected, and the diagnosis was further supported by a positive serum Leptospira interrogans icterohaemorrhagiae antibody titer. Unfortunately, he suffered from recurrent lower gastrointestinal bleeding, had a prolonged hospital course, and eventually succumbed to overwhelming sepsis. This case is the third report to our knowledge of leptospirosis in a renal transplant recipient in the English literature. PMID- 19558376 TI - Outcome of treatment of Epstein-Barr virus-related post-transplant lymphoproliferative disorder in hematopoietic stem cell recipients: a comprehensive review of reported cases. AB - Post-transplant lymphoproliferative disorder (PTLD) caused by Epstein-Barr virus (EBV) is an important complication in high-risk allogeneic hematopoietic stem cell transplant (HSCT) recipients. Before the current methods of anti-EBV therapy were introduced, the mortality from PTLD after HSCT was >80%. With current approaches the mortality from EBV-PTLD can be significantly reduced. The published literature and meeting abstracts were reviewed to assess the impact of different management strategies against EBV-PTLD. This analysis of reported outcomes indicates that preemptive use of rituximab and EBV-cytotoxic T lymphocytes (CTL) significantly reduced the risk of death due to EBV-PTLD in HSCT recipients with survival rates of 89.7% and 94.1%, respectively. Therapy of established PTLD also reduced the risk of fatal outcome. However, the overall success rates were lower than after preemptive therapy, reaching 63% and 88.2% of total EBV-DNA clearance with rituximab and CTL therapy, respectively. A reduction of immunosuppression and/or donor lymphocyte infusion might also reduce the risk of death due to EBV-PTLD. Although it is difficult to estimate these effects more precisely because of the frequent use of combination therapies, the responses to these modalities can be estimated to be 56.6% and 41.0%, respectively. Finally, chemotherapy seems not to contribute to improved survival of patients with PTLD after HSCT and antiviral agents are not active against PTLD. PMID- 19558377 TI - Heart rate recovery as a guide to monitor fatigue and predict changes in performance parameters. AB - Determining the optimal balance between training load and recovery contributes to peak performance in well-trained athletes. The measurement of heart rate recovery (HRR) to monitor this balance has become popular. However, it is not known whether the impairment in performance, which is associated with training-induced fatigue, is accompanied by a change in HRR. Therefore, the aim of this study was to retrospectively analyze the relationship between changes in HRR and cycling performance in a group of well-trained cyclists (n=14) who participated in a 4 week high-intensity training (HIT) program. Subjects were assigned to either a group that continuous had a increase in HRR (G(Incr)) or a group that showed a decrease in HRR (G(Decr)) during the HIT period. Both groups, G(Incr) and G(Decr), showed improvements in the relative peak power output (P=0.001 and 0.016, respectively) and endurance performance parameters (P=0.001 and <0.048, respectively). The average power during the 40-km time trial (40-km TT), however, improved more in G(Incr) (P=0.010), resulting in a tendency for a faster 40-km TT time (P=0.059). These findings suggest that HRR has the potential to monitor changes in endurance performance and contribute to a more accurate prescription of training load in well-trained and elite cyclists. PMID- 19558378 TI - Intrinsic risk factors for acute ankle injuries among male soccer players: a prospective cohort study. AB - This prospective cohort study was conducted to identify risk factors for acute ankle injuries among male soccer players. A total of 508 players representing 31 amateur teams were tested during the 2004 pre-season through a questionnaire on previous injury and function score (foot and ankle outcome score; FAOS), functional tests (balance tests on the floor and a balance mat) and a clinical examination of the ankle. Generalized estimating equations were used in univariate analyses to identify candidate risk factors, and factors with a P value <0.10 were then examined in a multivariate model. During the season, 56 acute ankle injuries, affecting 46 legs (43 players), were registered. Univariate analyses identified a history of previous acute ankle injuries [odds ratio (OR) per previous injury: 1.25, 95% confidence interval (CI) 1.09-1.43] and the FAOS sub-score "Pain" (OR for a 10-point difference in score: 0.81, 95% CI 0.62-1.04) as candidate risk factors. In a multivariate analysis, only the number of previous acute ankle injuries proved to be a significant (adjusted OR per previous injury: 1.23; 95% CI 1.06-1.41, P=0.005) predictor of new injuries. Function scores, functional tests and clinical examination could not independently identify players at an increased risk in this study. PMID- 19558379 TI - Transcriptional profiling: a potential anti-doping strategy. AB - Evolving challenges require evolving responses. The use of illicit performance enhancing drugs by athletes permeates the reality and the perception of elite sports. New drugs with ergogenic or masking potential are quickly adopted, driven by a desire to win and the necessity of avoiding detection. To counter this trend, anti-doping authorities are continually refining existing assays and developing new testing strategies. In the post-genome era, genetic- and molecular based tests are being evaluated as potential approaches to detect new and sophisticated forms of doping. Transcriptome analysis, in which a tissue's complement of mRNA transcripts is characterized, is one such method. The quantity and composition of a tissue's transcriptome is highly reflective of milieu and metabolic activity. There is much interest in transcriptional profiling in medical diagnostics and, as transcriptional information can be obtained from a variety of easily accessed tissues, similar approaches could be used in doping control. This article briefly reviews current understanding of the transcriptome, common methods of global analysis of gene expression and non-invasive sample sources. While the focus of this article is on anti-doping, the principles and methodology described could be applied to any research in which non-invasive, yet biologically informative sampling is desired. PMID- 19558380 TI - Facts, noise and wishful thinking: muscle protein turnover in aging and human disuse atrophy. AB - Surprisingly little is known about the mechanisms of muscle atrophy with aging and disuse in human beings, in contrast to rodents, from which much has been extrapolated to explain the human condition. However, this extrapolation is likely unwarranted because the time course, extent of wasting, muscle fiber involvement and alterations of muscle protein turnover are all quite different in rodent and human muscle. Furthermore, there is little evidence that static indices of protein turnover represent dynamic changes and may be misleading. With disuse there are reductions in the rate of muscle protein synthesis (MPS) large enough to explain the atrophic loss of muscle protein without a concomitant increase in proteolysis. In aging, there is no evidence that there are marked alterations in basal muscle protein turnover in healthy individuals but instead the ability to maintain muscle after feeding is compromised. This anabolic resistance is evident with physical inactivity, which exacerbates the inability to maintain muscle mass with aging. The main conclusion of this review is that in uncomplicated, non-inflammatory disuse atrophy, the facilitative change causing loss of muscle mass is a depression of MPS, exacerbated by anabolic resistance during feeding, with possible adaptive depressions, rather than increases, of muscle proteolysis. PMID- 19558381 TI - Effects of an anterior cruciate ligament injury prevention program on performance in adolescent female soccer players. AB - Female soccer players are three times more likely to suffer a non-contact anterior cruciate ligament (ACL) tear compared with male soccer players. Several ACL injury prevention programs have been developed and are used to reduce injury risk. However, to date there is limited information on how such programs affect physical performance. The aim of this randomized controlled study was to investigate the effects of the Prevent Injury Enhance Performance (PEP) program in adolescent female soccer players. Four soccer teams were randomly assigned to an intervention (PEP) or control (CON) group and assessed at baseline, 6 weeks, and 12 weeks on linear sprinting, countermovement jump (CMJ), and two agility tests. A mixed model factorial ANOVA with repeated measures was used to assess for treatment effects on the dependent variables. Improvements in 27.3 and 36.6 m sprint times (<0.10 s) were evident during the first 6 weeks for PEP, but reverted back to baseline values by 12 weeks; there were no changes for 9.1 or 18.2 m sprint times in either group. There was no change in the CMJ height for PEP; however, there was a decrement at 6 and 12 weeks compared with baseline in CON. Performance on the Illinois and pro-agility tests declined in both groups. Our findings demonstrate that improvements in linear sprint performance were small and transient in adolescent female soccer players, and that there was no benefit of the PEP program on CMJ or agility performance. ACL injury prevention programs designed as a structured warm-up routine seem to lack the necessary stimulus to enhance athletic performance. PMID- 19558382 TI - Regulation of extracellular matrix compounds involved in angiogenic processes in short- and long-track elite runners. AB - Exercise induces alterations of the extracellular matrix (ECM), e.g. by an increased release of endostatin or by regulation of matrix metalloproteases (MMP) 2/-9, and cathepsin L. To investigate the influence of training status on exercise-induced ECM-processing of angiogenic molecules, alterations of endostatin-, MMP-2, and MMP-9 plasma concentrations during incremental running step tests in male elite short-track (n=6) and male elite long-track runners (n=7) were studied. Three blood samples (pre-exercise, 0, and 1 h post-exercise) were taken from each subject at each running test. In both groups, the basal endostatin plasma concentration was significantly decreased at the second running test, i.e. after the training season. Exercise-related acute alterations of the parameters were also observed only during the second test. In the long-track group, there was a significant increase in endostatin at 0 h and of MMP-2 at 1 h post-exercise. In the short-track group, only MMP-9 was significantly increased at 0 h post-exercise. Cathepsin L was increased at 0 h post-exercise. In conclusion, regular exercise performance decreases the basal endostatin plasma concentration, facilitates ECM-processing of angiogenic molecules by regular performance, and seems to be dependent on the kind of training. PMID- 19558383 TI - Health-related fitness in adolescents: underweight, and not only overweight, as an influencing factor. The AVENA study. AB - This study investigated differences in health-related fitness (20-m shuttle run, handgrip, bent arm hang, standing long jump, shuttle run 4 x 10 m and sit and reach tests) in 2474 Spanish adolescents (1196 boys and 1278 girls; age 13-18.5 years) classed as underweight, normal weight, overweight or obese according to body mass index. Body fat and fat-free mass were derived from skinfold thickness. The prevalence of underweight was higher than obesity in girls (4.8% vs 3.0%, respectively; P<0.05) and the opposite in boys (3.9% vs 5.8%, respectively; P<0.05). Underweight was associated with a higher performance in the bent arm hang test in girls (P<0.05) and a lower performance in handgrip in both genders (P<0.01) compared with normal weight. Overweight and obese adolescents presented a lower performance in 20-m shuttle run, bent arm hang, standing long jump and shuttle run 4 x 10 m tests (P<0.001), but a higher performance in handgrip strength (P<0.001) compared with normal weight. In weight-bearing tests, the association became non-significant after adjusting for fat mass. In conclusion, not only overweight and obesity but also underweight seem to be determinants of health-related fitness in adolescents. The associations could be related to differences in body composition. PMID- 19558384 TI - Clinical assessment of hip strength using a hand-held dynamometer is reliable. AB - Hip strength assessment plays an important role in the clinical examination of the hip and groin region. The primary aim of this study was to examine the absolute test-retest measurement variation concerning standardized strength assessments of hip abduction (ABD), adduction (ADD), external rotation (ER), internal rotation (IR), flexion (FLEX) and extension (EXT) using a hand-held dynamometer. Nine subjects (five males, four females), physically active for at least 2.5 h a week, were included. Twelve standardized isometric strength tests were performed twice with a 1-week interval in between by the same examiner. The test order was randomized to avoid systematic bias. Measurement variation between sessions was 3-12%. When the maximum value of four measurements was used, test retest measurement variation was below 10% in 11 of the 12 individual hip strength tests and below 5% in five of the 12 tests. No systematic differences were present. Standardized strength assessment procedures of hip ABD, ER, IR, FLEX, with test-retest measurement variation below 5%, hip ADD below 6% and hip EXT below 8%, make it possible to determine even small changes in hip strength at the individual level. PMID- 19558385 TI - Calculation of the contact pressure between ski and snow during a carved turn in Alpine skiing. AB - The macroscopic contact area between ski and snow and the contact pressure are crucial influencing factors for carved turns in Alpine skiing. In the present paper, a simulation model is developed to quantify these factors. The ski is modelled as an Euler-Bernoulli beam with variable cross section, camber, bending and torsional stiffness using measured data from skis. The reaction forces of the snow are decomposed in penetration and shear forces. For the penetration forces a hypoplastic constitutive law is applied incorporating elastic and plastic deformation of the snow at the contact area. For the shear forces metal cutting theory is used. Ski deformation, contact area and contact pressure are computed based on quasi-static equilibrium between forces exerted by the skier and snow reaction forces. Parameter studies are performed to investigate the influence of edging and distributing the load between the inner and outer ski. Higher edging angles as well as loading both skis affected the contact pressure positively by increasing the resistance against shearing. The results of our study agree well with measurement data taken from literature. Based on the results, the importance of actions of the skier during carved turns is concluded. PMID- 19558386 TI - Thermoregulatory demands of elite professional America's Cup yacht racing. AB - America's Cup yacht racing predominantly occurs during the summer months under hot and humid conditions, with athletes exposed to the environment for prolonged periods, and yet the thermoregulatory responses to competitive sailing are largely unappreciated. This study aimed to assess the thermoregulatory responses to elite professional big-boat yacht racing, according to crew position and upwind and downwind sailing. Intestinal (T(core)) and skin temperature, fluid balance and regional sweat compositions were measured in two America's Cup crews (n=32) during 100 min of racing. The environmental conditions were as follows: 32 degrees C, 52% RH and 5 m/s wind speed. Subjective race intensity was moderate. Bowmen recorded the greatest elevation in the heart rate (184 +/- 10 beats/min) and T(core) (39.2 degrees C, P<0.01). Both heart rate and T(core) were higher during downwind sailing (P<0.001). Regional skin temperatures were significantly different according to site (P=0.05), with tibia being the lowest (33.3 +/- 1.2 degrees C). The mean sweat loss during racing was 1.34 +/- 0.58 L/h (range: 0.44 2.40 L/h), with bowmen experiencing the greatest loss of sweat (3.7 +/- 0.9% of body mass). The mean fluid intake was highly correlated to sweat loss (r=0.74, P<0.001), with 72 +/- 41% of sweat losses replaced. The mean sodium concentration of sweat was 27.2 +/- 9.2 mmol/L (range: 12.0-43.5 mmol/L) and the total NaCl loss during sailing was 3.8 +/- 2.4 g (range 0.7-10.0 g). America's Cup sailing is a demanding sport that presents considerable challenges to thermoregulation, fluid and electrolyte balance. Certain crew roles (bowmen) present an increased risk of developing exertional heat illness, and for the majority of crew downwind sailing results in greater thermal strain than upwind sailing - which may have implications for clothing selection and boat design. PMID- 19558387 TI - Inspiratory flow resistive loading improves respiratory muscle function and endurance capacity in recreational runners. AB - The purpose of this study was to assess the efficacy of inspiratory flow resistive loading (IFRL) on respiratory muscle function, exercise performance and cardiopulmonary and metabolic responses to exercise. Twenty-four recreational road runners (12 male) were randomly assigned from each gender into an IFRL group (n=8) and sham-IFRL group (n=8), which performed IFRL for 6 weeks, or a control group (n=8). Strength (+43.9%Delta), endurance (+26.6%Delta), maximum power output (+41.9%Delta) and work capacity (+38.5%Delta) of the inspiratory muscles were significantly increased (P<0.05) at rest following the study period in IFRL group only. In addition, ventilation (-25.7%Delta), oxygen consumption ( 13.3%Delta), breathing frequency (-11.9%Delta), tidal volume (-16.0%Delta), heart rate (HR) (-13.1%Delta), blood lactate concentration (-38.9%Delta) and the perceptual response (-33.5%Delta) to constant workload exercise were significantly attenuated (P<0.05), concomitant with a significant improvement (P<0.05) in endurance exercise capacity (+16.4%Delta) during a treadmill run set at 80% VO2max in IFRL group only. These data suggest that IFRL can alter breathing mechanics, attenuate the oxygen cost, ventilation, HR, blood lactate and the perceptual response during constant workload exercise and improve endurance exercise performance in recreational runners. PMID- 19558388 TI - Epidemiologic evidence for chloroprene carcinogenicity: review of study quality and its application to risk assessment. AB - This article evaluates the quality and weight of evidence associated with epidemiologic studies of cancer among occupational cohorts exposed to chloroprene. The focus is on liver, lung, and lymphohematopoietic cancers, which had been increased in early studies. Literature searches identified eight morbidity/mortality studies covering seven chloroprene-exposed cohorts from six countries. These studies were summarized and their quality was assessed using the 10 criteria suggested by the U.S. Environmental Protection Agency. The limitations within this literature (primarily the early studies) included crude exposure assessment, incomplete follow-up, uncertain baseline rates, and uncontrolled confounding by factors such as smoking, drinking, and co-exposure to benzene and vinyl chloride. Four cohorts were studied by the same group of investigators, who reported no overall increased associations for any cancers. This four-cohort study was by far the most rigorous, having the most comprehensive exposure assessment and follow-up and the most detailed documentation. This study also contained the two largest cohorts, including an American cohort from Louisville, Kentucky, that ranked at or near the top for each of the 10 quality criteria. There was evidence of a strong healthy worker effect in the four-cohort study, which could have hidden small excess risks. Small increased risks were suggested by internal or company-specific analyses, but these were most likely caused by uncontrolled confounding and low baseline rates. Overall, the weight of evidence does not support any substantial link between chloroprene exposure and cancer, but inconsistencies and a lack of control for major confounders preclude drawing firmer conclusions. PMID- 19558389 TI - Relative contributions of four exposure pathways to influenza infection risk. AB - The relative contribution of four influenza virus exposure pathways-(1) virus contaminated hand contact with facial membranes, (2) inhalation of respirable cough particles, (3) inhalation of inspirable cough particles, and (4) spray of cough droplets onto facial membranes-must be quantified to determine the potential efficacy of nonpharmaceutical interventions of transmission. We used a mathematical model to estimate the relative contributions of the four pathways to infection risk in the context of a person attending a bed-ridden family member ill with influenza. Considering the uncertainties in the sparse human subject influenza dose-response data, we assumed alternative ratios of 3,200:1 and 1:1 for the infectivity of inhaled respirable virus to intranasally instilled virus. For the 3,200:1 ratio, pathways (1), (2), and (4) contribute substantially to influenza risk: at a virus saliva concentration of 10(6) mL(-1), pathways (1), (2), (3), and (4) contribute, respectively, 31%, 17%, 0.52%, and 52% of the infection risk. With increasing virus concentrations, pathway (2) increases in importance, while pathway (4) decreases in importance. In contrast, for the 1:1 infectivity ratio, pathway (1) is the most important overall: at a virus saliva concentration of 10(6) mL(-1), pathways (1), (2), (3), and (4) contribute, respectively, 93%, 0.037%, 3.3%, and 3.7% of the infection risk. With increasing virus concentrations, pathway (3) increases in importance, while pathway (4) decreases in importance. Given the sparse knowledge concerning influenza dose and infectivity via different exposure pathways, nonpharmaceutical interventions for influenza should simultaneously address potential exposure via hand contact to the face, inhalation, and droplet spray. PMID- 19558390 TI - Does communicating (flood) risk affect (flood) risk perceptions? Results of a quasi-experimental study. AB - People's risk perceptions are generally regarded as an important determinant of their decisions to adjust to natural hazards. However, few studies have evaluated how risk communication programs affect these risk perceptions. This study evaluates the effects of a small-scale flood risk communication program in the Netherlands, consisting of workshops and focus group discussions. The effects on the workshop participants' (n = 24) and focus group participants' (n = 16) flood risk perceptions were evaluated in a pretest-posttest control group (n = 40) design that focused on two mechanisms of attitude change-direct personal experience and attitude polarization. We expected that (H1) workshop participants would show greater shifts in their flood risk perceptions compared with control group participants and that (H2) focus groups would rather produce the conditions for attitude polarization (shifts toward more extreme attitudinal positions after group discussion). However, the results provide only modest support for these hypotheses, perhaps because of a mismatch between the sessions' contents and the risk perception measures. An important contribution of this study is that it examined risk perception data by both conventional tests of the mean differences and tests for attitude polarization. Moreover, the possibility that attitude polarization could cause people to confirm their preexisting (hazard) beliefs could have important implications for risk communication. PMID- 19558391 TI - Evaluating critical uncertainty thresholds in a spatial model of forest pest invasion risk. AB - Pest risk maps can provide useful decision support in invasive species management, but most do not adequately consider the uncertainty associated with predicted risk values. This study explores how increased uncertainty in a risk model's numeric assumptions might affect the resultant risk map. We used a spatial stochastic model, integrating components for entry, establishment, and spread, to estimate the risks of invasion and their variation across a two dimensional landscape for Sirex noctilio, a nonnative woodwasp recently detected in the United States and Canada. Here, we present a sensitivity analysis of the mapped risk estimates to variation in key model parameters. The tested parameter values were sampled from symmetric uniform distributions defined by a series of nested bounds (+/-5%, ... , +/-40%) around the parameters' initial values. The results suggest that the maximum annual spread distance, which governs long distance dispersal, was by far the most sensitive parameter. At +/-15% or larger variability bound increments for this parameter, there were noteworthy shifts in map risk values, but no other parameter had a major effect, even at wider bounds of variation. The methodology presented here is generic and can be used to assess the impact of uncertainties on the stability of pest risk maps as well as to identify geographic areas for which management decisions can be made confidently, regardless of uncertainty. PMID- 19558394 TI - Hands-free embedded cell phones and airbag-deployment crash rates. PMID- 19558396 TI - The elephant in the room is called game theory. PMID- 19558397 TI - Empirical analysis of the effects of cyber security incidents. AB - We analyze the time series associated with web traffic for a representative set of online businesses that have suffered widely reported cyber security incidents. Our working hypothesis is that cyber security incidents may prompt (security conscious) online customers to opt out and conduct their business elsewhere or, at the very least, to refrain from accessing online services. For companies relying almost exclusively on online channels, this presents an important business risk. We test for structural changes in these time series that may have been caused by these cyber security incidents. Our results consistently indicate that cyber security incidents do not affect the structure of web traffic for the set of online businesses studied. We discuss various public policy considerations stemming from our analysis. PMID- 19558398 TI - The stability of error-related brain activity with increasing trials. AB - The error-related negativity (ERN) and error positivity (Pe) are increasingly being examined as neural correlates of response monitoring. The minimum number of error trials included in grand averages varies across studies; indeed, there has not been a systematic investigation on the number of trials required to obtain a stable ERN and Pe. In the current study, the ERN and Pe were quantified as two random trials were added to participants' (N=53) ERP averages. Adding trials increased the correlation with the grand average ERN and Pe; however, high correlations (rs>.80) were obtained with only 6 trials. Internal reliability of the ERN and Pe reached moderate levels after 6 and 2 trials and the signal-to noise ratio of the ERN and Pe did not change after 8 and 4 trials, respectively. Combined, these data suggest that the ERN and Pe can be quantified using a minimum of between 6 and 8 error trials. PMID- 19558399 TI - Enhanced cardiac perception is associated with benefits in decision-making. AB - In the present study we provide the first empirical evidence that viscero-sensory feedback from an internal organ is associated with decision-making processes. Participants with accurate vs. poor perception of their heart activity were compared with regard to their performance in the Iowa Gambling Task. During this task, participants have to choose between four card decks. Decks A and B yield high gains and high losses, and if played continuously, result in net loss. In contrast, decks C and D yield small gains and also small losses, but result in net profit if they are selected continuously. Accordingly, participants have to learn to avoid the net loss options in favor of the net gain options. In our study, participants with good cardiac perception chose significantly more of the net gain and fewer of the net loss options. Our findings document the substantial role of visceral feedback in decision-making processes in complex situations. PMID- 19558400 TI - Aging effects on early-stage face perception: an ERP study. AB - We compared early stages of face processing in young and older participants as indexed by ERPs elicited by faces and non-face stimuli presented in upright and inverted orientations. The P1 and N170 components were larger in older than in young participants. However, the early distinction between stimulus categories as reflected by N170 face was similar across groups. Face inversion increased and delayed the N170 peak in the younger group while in older participants inversion delayed the N170 peak but had no effect on amplitude. The N170 amplitude was right-lateralized in the young, but not in the older group. Yet, the difference between the N170 elicited by faces and non-face stimuli was similarly right lateralized in both groups. These data suggest that detection of faces and their streaming to face-characteristic structural encoding is not altered by age. In contrast, the absence of face-inversion effects on N170 amplitudes in the elderly suggest that face individuation, which is probably the default strategy in younger people, might not be attempted by default in older people, at least when they look at young faces. PMID- 19558401 TI - An alternative scoring method for skin conductance responding in a differential fear conditioning paradigm with a long-duration conditioned stimulus. AB - Researchers examining skin conductance (SC) as a measure of aversive conditioning commonly separate the SC response into two components when the CS-UCS interval is sufficiently long. This convention drew from early theorists who described these components, the first- and second-interval responses, as measuring orienting and conditional responses, respectively. The present report critically examines this scoring method through a literature review and a secondary data analysis of a large-scale study of police and firefighter trainees that used a differential aversive conditioning procedure (n=287). The task included habituation, acquisition, and extinction phases, with colored circles as the CSs and shocks as the UCS. Results do not support the convention of separating the SC response into first- and second-interval responses. It is recommended that SC response scores be derived from data obtained across the entire CS-UCS interval. PMID- 19558403 TI - Theoretical reconsiderations when estimating the mesophyll conductance to CO(2) diffusion in leaves of C(3) plants by analysis of combined gas exchange and chlorophyll fluorescence measurements. AB - Existing methods to estimate the mesophyll conductance to CO(2) diffusion (g(m)) are often based on combined gas exchange and chlorophyll fluorescence measurements. However, estimations of average g(m) by these methods are often unreliable either because the range of usable data is too narrow or because the estimations are very sensitive to measurement errors. We describe three method variants to estimate g(m), for which a wider range of data are usable. They use curve-fitting techniques, which minimise the sum of squared model deviations from the data for A (CO(2) assimilation rate) or for J (linear electron transport rate). Like the existing approaches, they are all based on common physiological principles assuming that electron transport limits A. The proposed variants were far less sensitive than the existing approaches to 'measurement noise' either created randomly in the generated data set or inevitably existing in real data sets. Yet, the estimates of g(m) from the three variants differed by approximately 15%. Moreover, for each variant, a stoichiometric uncertainty in linear electron transport-limited photosynthesis can cause another 15% difference. Any estimation of g(m) using gas exchange and chlorophyll fluorescence measurements should be considered with caution, especially when g(m) is high. PMID- 19558404 TI - Thermogenesis of three species of Arum from Crete. AB - Inflorescences of arum lilies have a three-part spadix with a scent-producing, sterile appendix above two bands of fertile male and female florets. The appendix and male florets are thermogenic, but with different temporal patterns. Heat production was measured in Arum concinnatum, A. creticum and A. idaeum. The male florets of A. concinnatum showed a 3 d continuous episode of thermogenesis with three waves, and the appendix warmed in a single, 6 h episode. Maximum fresh-mass specific CO(2) production rate was 0.17 micromol s(-1) g(-1) to achieve a 10.9 degrees C temperature elevation by the appendix, and 0.92 micromol s(-1) g(-1) to achieve a 4.8 degrees C elevation by male florets. Reversible, physiological temperature regulation was not evident in either tissue. Respiration increased with tissue temperatures with Q(10) values of 1.8-3.9, rather than less than 1.0 as occurs in thermoregulatory flowers. Experimental step changes in temperature of appendix and male floret tissues also failed to show thermoregulatory responses. The patterns of thermogenesis therefore appear to be fixed by the temporal sequence of blooming. Thermogenesis in the alpine species, A. creticum and A. idaeum, was significantly lower than in the lowland A. concinnatum, possibly related to difficulty in raising floral temperature in their cold and windy habitat. PMID- 19558402 TI - The heritability of P300 amplitude in 18-year-olds is robust to adolescent alcohol use. AB - P3 amplitude reduction (P3AR) is associated with adolescent alcohol use (AAU) and highly heritable, suggesting that P3AR may index a genetic predisposition (e.g., an endophenotype) for AAU. However, because P3AR and AAU covary naturally in the population, these observations are also consistent with P3AR reflecting neurotoxic effects of AAU on the developing adolescent brain. In this report, we describe the use of recent advancements in biometric modeling to examine changes in the genetic and environmental contributions to variability in P3 amplitude related to cumulative AAU by late adolescence in a large community-based twin sample. We found that the genetic and environmental contributions to variability in P3 amplitude were unaffected by AAU. This suggests that P3AR indexes risk for alcoholism independent of any deleterious effect of AAU on adolescent brain development. PMID- 19558405 TI - Acclimation of leaf hydraulic conductance and stomatal conductance of Pinus taeda (loblolly pine) to long-term growth in elevated CO(2) (free-air CO(2) enrichment) and N-fertilization. AB - We investigated how leaf hydraulic conductance (K(leaf)) of loblolly pine trees is influenced by soil nitrogen amendment (N) in stands subjected to ambient or elevated CO(2) concentrations (CO(2)(a) and CO(2)(e), respectively). We also examined how K(leaf) varies with changes in reference leaf water potential (Psi(leaf-ref)) and stomatal conductance (g(s-ref)) calculated at vapour pressure deficit, D of 1 kPa. We detected significant reductions in K(leaf) caused by N and CO(2)(e), but neither treatment affected pre-dawn or midday Psi(leaf). We also detected a significant CO(2)(e)-induced reduction in g(s-ref) and Psi(leaf ref). Among treatments, the sensitivity of K(leaf) to Psi(leaf) was directly related to a reference K(leaf) (K(leaf-ref) computed at Psi(leaf-ref)). This liquid-phase response was reflected in a similar gas-phase response, with g(s) sensitivity to D proportional to g(s-ref). Because leaves represented a substantial component of the whole-tree conductance, reduction in K(leaf) under CO(2)(e) affected whole-tree water use by inducing a decline in g(s-ref). The consequences of the acclimation of leaves to the treatments were: (1) trees growing under CO(2)(e) controlled morning leaf water status less than CO(2)(a) trees resulting in a higher diurnal loss of K(leaf); (2) the effect of CO(2)(e) on g(s-ref) was manifested only during times of high soil moisture. PMID- 19558406 TI - Oxygen isotope enrichment (Delta(18)O) reflects yield potential and drought resistance in maize. AB - Measurement of stable isotopes in plant dry matter is a useful phenotypic tool for speeding up breeding advance in C(3) crops exposed to different water regimes. However, the situation in C(4) crops is far from resolved, since their photosynthetic metabolism precludes (at least in maize) the use of carbon isotope discrimination. This paper investigates the use of oxygen isotope enrichment (Delta(18)O) as a new secondary trait for yield potential and drought resistance in maize (Zea mays L). A set of tropical maize hybrids developed by the International Maize and Wheat Improvement Center was grown under three contrasting water regimes in field conditions. Water regimes clearly affected plant growth and yield. In accordance with the current theory, a decrease in water input was translated into large decreases in stomatal conductance and increases in leaf temperature together with concomitant (18)O enrichment of plant matter (leaves and kernels). In addition, kernel Delta(18)O correlated negatively with grain yield under well-watered and intermediate water stress conditions, while it correlated positively under severe water stress conditions. Therefore, genotypes showing lower kernel Delta(18)O under well-watered and intermediate water stress had higher yields in these environments, while the opposite trend was found under severe water stress conditions. This illustrates the usefulness of Delta(18)O for selecting the genotypes best suited to differing water conditions. PMID- 19558407 TI - Size-dependent mortality in a Neotropical savanna tree: the role of height related adjustments in hydraulic architecture and carbon allocation. AB - Size-related changes in hydraulic architecture, carbon allocation and gas exchange of Sclerolobium paniculatum (Leguminosae), a dominant tree species in Neotropical savannas of central Brazil (Cerrado), were investigated to assess their potential role in the dieback of tall individuals. Trees greater than approximately 6-m-tall exhibited more branch damage, larger numbers of dead individuals, higher wood density, greater leaf mass per area, lower leaf area to sapwood area ratio (LA/SA), lower stomatal conductance and lower net CO(2) assimilation than small trees. Stem-specific hydraulic conductivity decreased, while leaf-specific hydraulic conductivity remained nearly constant, with increasing tree size because of lower LA/SA in larger trees. Leaves were substantially more vulnerable to embolism than stems. Large trees had lower maximum leaf hydraulic conductance (K(leaf)) than small trees and all tree sizes exhibited lower K(leaf) at midday than at dawn. These size-related adjustments in hydraulic architecture and carbon allocation apparently incurred a large physiological cost: large trees received a lower return in carbon gain from their investment in stem and leaf biomass compared with small trees. Additionally, large trees may experience more severe water deficits in dry years due to lower capacity for buffering the effects of hydraulic path-length and soil water deficits. PMID- 19558408 TI - Root based approaches to improving nitrogen use efficiency in plants. AB - In the majority of agricultural growing regions, crop production is highly dependent on the supply of exogenous nitrogen (N) fertilizers. Traditionally, this dependency and the use of N-fertilizers to restore N depleted soils has been rewarded with increased plant health and yields. In recent years, increased competition for non-renewable fossil fuel reserves has directly elevated prices of N-fertilizers and the cost of agricultural production worldwide. Furthermore, N-fertilizer based pollution is becoming a serious issue for many regions where agriculture is highly concentrated. To help minimize the N footprint associated with agricultural production there is significant interest at the plant level to develop technologies which can allow economically viable production while using less applied N. To complement recent reviews examining N utilization efficiency in agricultural plants, this review will explore those strategies operating specifically at the root level, which may directly contribute to improved N use efficiencies in agricultural crops such as cereals, where the majority of N fertilizers are used and lost to the environment. Root specific phenotypes that will be addressed in the context of improvements to N acquisition and assimilation efficiencies include: root morphology; root to shoot ratios; root vigour, root length density; and root N transport and metabolism. PMID- 19558409 TI - Physiological and isotopic (delta(13)C and delta(18)O) responses of three tropical tree species to water and nutrient availability. AB - Water-use efficiency and stable isotope composition were studied in three tropical tree species. Seedlings of Tectona grandis, Swietenia macrophylla and Platymiscium pinnatum were grown at either high or low water supply, and with or without added fertilizer. These three species previously exhibited low, intermediate and high whole-plant water-use efficiency (TE) when grown at high water supply in unfertilized soil. Responses of TE to water and nutrient availability varied among species. The TE was calculated as experiment-long dry matter production divided by cumulative water use. Species-specific offsets were observed in relationships between TE and whole-plant (13)C discrimination (Delta(13)C(p)). These offsets could be attributed to a breakdown in the relationship between Delta(13)C(p) and the ratio of intercellular to ambient CO(2) partial pressures (c(i)/c(a)) in P. pinnatum, and to variation among species in the leaf-to-air vapour pressure difference (v). Thus, a plot of v.TE against c(i)/c(a) showed a general relationship among species. Relationships between delta(18)O of stem dry matter and stomatal conductance ranged from strongly negative for S. macrophylla to no relationship for T. grandis. Results suggest inter-specific variation among tropical tree species in relationships between stable isotope ratios (delta(13)C and delta(18)O) and the gas exchange processes thought to affect them. PMID- 19558410 TI - Adaptation of plasma membrane H(+)-ATPase of rice roots to low pH as related to ammonium nutrition. AB - The preference of paddy rice for NH(4)(+) rather than NO(3)(-) is associated with its tolerance to low pH since a rhizosphere acidification occurs during NH(4)(+) absorption. However, the adaptation of rice root to low pH has not been fully elucidated. This study investigated the acclimation of plasma membrane H(+) ATPase of rice root to low pH. Rice seedlings were grown either with NH(4)(+) or NO(3)(-). For both nitrogen forms, the pH value of nutrient solutions was gradually adjusted to pH 6.5 or 3.0. After 4 d cultivation, hydrolytic H(+) ATPase activity, V(max), K(m), H(+)-pumping activity, H(+) permeability and pH gradient across the plasma membrane were significantly higher in rice roots grown at pH 3.0 than at 6.5, irrespective of the nitrogen forms supplied. The higher activity of plasma membrane H(+)-ATPase of adapted rice roots was attributed to the increase in expression of OSA1, OSA3, OSA7, OSA8 and OSA9 genes, which resulted in an increase of H(+)-ATPase protein concentration. In conclusion, a high regulation of various plasma membrane H(+)-ATPase genes is responsible for the adaptation of rice roots to low pH. This mechanism may be partly responsible for the preference of rice plants to NH(4)(+) nutrition. PMID- 19558411 TI - OsMADS50 and OsMADS56 function antagonistically in regulating long day (LD) dependent flowering in rice. AB - In much of the tropics and subtropics, rice (Oryza sativa L.) is grown under long days (LDs). Therefore, LD must play a major role in inducing flowering signal in rice. However, little is known on LD-dependent flowering signal in the species. We previously reported that OsMADS50, which is highly homologous to Arabidopsis SOC1, functions as a positive regulator for flowering. However, its detailed photoperiodic mechanism was not yet elucidated. Here, we report the functional analysis of OsMADS50 and its closely related gene OsMADS56. Knock-out of OsMADS50 caused a late-flowering phenotype only under LD conditions. Overexpression of OsMADS56 (56OX) also resulted in delayed flowering under LD. In the osmads50 mutants and 56OX transgenic plants, transcripts of Ehd1, Hd3a and RFT1 were reduced, although that of OsLFL1 increased. On the other hand, mRNA levels of OsGI, Hd1, OsId1, OsDof12, Ghd7, Hd6 and SE5 were unchanged. These observations imply that OsMADS50 and OsMADS56 function antagonistically through OsLFL1-Ehd1 in regulating LD-dependent flowering. Yeast two-hybrid and co-immunoprecipitation analyses indicated an interaction between those two proteins as well as their formation of homodimers. These results suggest that OsMADS50 and OsMADS56 may form a complex that regulates downstream target genes. PMID- 19558412 TI - Diurnal and seasonal variation in the carbon isotope composition of leaf dark respired CO(2) in velvet mesquite (Prosopis velutina). AB - We evaluated diurnal and seasonal patterns of carbon isotope composition of leaf dark-respired CO(2) (delta(13)C(l)) in the C(3) perennial shrub velvet mesquite (Prosopis velutina) across flood plain and upland savanna ecosystems in the south western USA. delta(13)C(l) of darkened leaves increased to maximum values late during daytime periods and declined gradually over night-time periods to minimum values at pre-dawn. The magnitude of the diurnal shift in delta(13)C(l) was strongly influenced by seasonal and habitat-related differences in soil water availability and leaf surface vapour pressure deficit. delta(13)C(l) and the cumulative flux-weighted delta(13)C value of photosynthates were positively correlated, suggesting that progressive (13)C enrichment of the CO(2) evolved by darkened leaves during the daytime mainly resulted from short-term changes in photosynthetic (13)C discrimination and associated shifts in the delta(13)C signature of primary respiratory substrates. The (13)C enrichment of dark respired CO(2) relative to photosynthates across habitats and seasons was 4 to 6 per thousand at the end of the daytime period (1800 h), but progressively declined to 0 per thousand by pre-dawn (0300 h). The origin of night-time and daytime variations in delta(13)C(l) is discussed in terms of the carbon source(s) feeding respiration and the drought-induced changes in carbon metabolism. PMID- 19558413 TI - Metabolomic and physiological responses reveal multi-phasic acclimation of Arabidopsis thaliana to chronic UV radiation. AB - Biochemical changes in vivo and pathway interactions were investigated using integrated physiological and metabolic responses of Arabidopsis thaliana L. to ultraviolet (UV) radiation (280-400 nm) at 9.96 kJ m(-2) d(-1) over the entire life cycle from seed to seed (8 weeks). Columbia-0 (Col-0) and a UV-B sensitive accession (fah-1) showed significant (P < 0.001) reductions in leaf growth after 6 weeks. Col-0 recovered growth after 8 weeks, with recovery corresponding to a switch from production of phenylpropanoids to flavonoids. fah-1 failed to recover, indicating that sinapate production is an essential component of recovery. Epidermal features show that UV radiation caused significant (P < 0.001) increases in trichome density, which may act as a structural defence response. Stomatal indices showed a significant (P < 0.0001) reduction in Col-0 and a significant (P < 0.001) increase in fah-1. Epidermal cell density was significantly increased under UV radiation on the abaxial leaf surface, suggesting that that a fully functioning phenylpropanoid pathway is a requirement for cell expansion and leaf development. Despite wild-type acclimation, the costs of adaptation lead to reduced plant fitness by decreasing flower numbers and total seed biomass. A multi-phasic acclimation to UV radiation and the induction of specific metabolites link stress-induced biochemical responses to enhanced acclimation. PMID- 19558414 TI - Public health issues arising from sale of illegal and potentially unsafe 'tanning chemicals'. PMID- 19558415 TI - alpha-MSH tripeptide analogs activate the melanocortin 1 receptor and reduce UV induced DNA damage in human melanocytes. AB - One skin cancer prevention strategy that we are developing is based on synthesizing and testing melanocortin analogs that reduce and repair DNA damage resulting from exposure to solar ultraviolet (UV) radiation, in addition to stimulating pigmentation. Previously, we reported the effects of tetrapeptide analogs of alpha-melanocortin (alpha-MSH) that were more potent and stable than the physiological alpha-MSH, and mimicked its photoprotective effects against UV induced DNA damage in human melanocytes. Here, we report on a panel of tripeptide analogs consisting of a modified alpha-MSH core His(6)-d-Phe(7)-Arg(8), which contained different N-capping groups, C-terminal modifications, or arginine mimics. The most potent tripeptides in activating cAMP formation and tyrosinase of human melanocytes were three analogs with C-terminal modifications. The most effective C-terminal tripeptide mimicked alpha-MSH in reducing hydrogen peroxide generation and enhancing nucleotide excision repair following UV irradiation. The effects of these three analogs required functional MC1R, as they were absent in human melanocytes that expressed non-functional receptor. These results demonstrate activation of the MC1R by tripeptide melanocortin analogs. Designing small analogs for topical delivery should prove practical and efficacious for skin cancer prevention. PMID- 19558416 TI - Alternative oxidase: a defence against metabolic fluctuations? AB - An increasing number of oscillating or fluctuating cellular systems have been recently described following the adaptation of fluorescent technology. In diverse organisms, these variously involve signalling factors, heat production, central metabolism and reactive oxygen species (ROS). In response to many plant stresses and primarily via the influence of ROS, changes in mRNA and protein levels or in vivo activity of alternative oxidase are often observed. However, in several investigations, a lack of correlation between the mRNA, protein and in vivo activity has been evident. This discrepancy has made it questionable whether the induction of alternative oxidase has importance in regulating alternative pathway activity in vivo, or being diagnostic for a role of alternative oxidase in stress tolerance and ROS avoidance. Here, we suggest a role of alternative oxidase in counteracting deleterious short-term metabolic fluctuations, especially under stress conditions. This model emphasizes the importance of peak activity for establishing protein levels and allows an amalgamation of the present status of physiological, cellular and molecular knowledge. PMID- 19558417 TI - Limited proteolysis of luciferase as a reporter in nanosystem biology: a comparative study. AB - Firefly luciferase is a 62 kDa protein that produces a flash of light on the oxidation of luciferin in the presence of ATP, Oxygen and Mg2+. Luciferase has a broad range of applications for analytical purposes and in vivo imaging for nanosystem biology studies. However, the enzyme is highly susceptible to proteolytic degradation that reduces its half-life. Rate of proteolytic digestion between two members of luciferase family (Photinus pyralis and Lampyris turkestanicus) is compared. Proteolytic sensitivity of L. turkestanicus luciferase was found to be more than P. pyralis luciferase, due to higher rate of hydrolysis under identical conditions. Both luciferases showed more sensitivity to chymotrypsin than trypsin with different digestion pattern. Digestion of P. pyralis by trypsin produced some fragments which were found to be more resistant to further degradation, whereas in L. turkestanicus initial fragments subdigested by trypsin, like chymotrypsin effect on both luciferases. Furthermore, both luciferases become increasingly labile to proteolysis as the temperature increases. The rate of inactivation and the rate of degradation between luciferases were different in a specific time of incubation. Appearance of similar bands for both luciferases confirmed exposure of specific regions, in spite of structural differences. PMID- 19558418 TI - Dye-surfactant interaction: modulation of photophysics of an ionic styryl dye. AB - The interesting modulation of multibond rotation-induced intramolecular charge transfer photophysics of 2-(4-(dimethylamino) styryl)-1-methylpyridinium iodide in different micelles due to different contributions of twisted intramolecular charge transfer (TICT) and hydrogen bonding deactivation channels have been reported in this paper. 2-(4-(dimethylamino) styryl)-1-methylpyridinium iodide enters into all the micelles in different positions from the water solution due to active hydrophobic force and electrostatic field, as revealed from the shift and intensity of intramolecular charge transfer (ICT) band. The presence of mechanically trapped water with the addition of salt and inherent thermodynamic water controls the ICT emission. Analysis of spectral data before and after the addition of salt confirms the orientation of 2-(4-(dimethylamino) styryl)-1 methylpyridinium iodide in cationic and anionic micelles. PMID- 19558419 TI - Laser selection significantly affects cell viability following single-cell nanosurgery. AB - This paper compares the viability of over 700 NG108 cells after membrane disruption either with a single 3 ns pulse at 337 nm or with a 5 ms train of 110 fs pulses (80 MHz) at 770 nm. Cell viability was monitored over a period of 12 h so as to understand the effect of laser ablation-induced cell apoptosis. The use of one-photon membrane disruption with the UV-laser resulted in approximately 36% cell viability after 12 h while the use of two-photon ablation with the femtosecond laser resulted in a much higher viability of approximately 79% after 12 h, which was the same within error of the approximately 79% viability of cells in the control group. Changing the laser power to achieve a 90% probability of membrane disruption (PMD) from 50% PMD did not change the percentage of viable cells after 12 h, regardless of whether one- or two-photon ablation was employed. A systematic comparison between different methods of cellular ablation and their effect upon the viability of single cells has not been done before over such a long time frame. These results show the importance of laser choice when cell viability postsurgery is a concern. PMID- 19558420 TI - Physicochemical studies of demetalation of light-harvesting bacteriochlorophyll isomers purified from green sulfur photosynthetic bacteria. AB - Demetalation kinetics of bacteriochlorophylls (BChls) c, d and e from green sulfur photosynthetic bacteria were studied under weakly acidic conditions. Demetalation rate constants of BChl e possessing a formyl group at the 7-position were significantly smaller than those of BChls c and d, which had a methyl group at this position. The activation energy of demetalation of 3(1)R-8,12 diethyl([E,E])-BChl e was 1.5-times larger than that of 3(1)R-[E,E]-BChl c. 15N labeled 3(1)R-[E,E]-BChls c and e were purified from cells of green sulfur bacteria grown in a medium containing 15NH4Cl, and their 15N NMR spectra were measured. The chemical shifts of N21, N22 and N23 atoms of 3(1)R-[E,E]-BChl e were lower-field shifted than those of 3(1)R-[E,E]-BChl c, respectively, and especially the difference in chemical shifts of N22 was significantly large. These results suggest that the electron-withdrawing formyl group at the 7 position of BChl e affected an electronic state of the chlorin macrocycle and caused BChl e to be more tolerant for removal of the central magnesium compared with BChls c and d. PMID- 19558421 TI - UV-B action spectrum for UVR8-mediated HY5 transcript accumulation in Arabidopsis. AB - Arabidopsis thaliana UV RESISTANCE LOCUS8 (UVR8) is a UV-B-specific signaling component that mediates low fluence, photomorphogenic responses to UV-B. It is required for UV-B-induced expression of the gene encoding the ELONGATED HYPOCOTYL5 (HY5) transcription factor. HY5 is a key effector of responses mediated by UVR8. In mature leaf tissue, HY5 transcript accumulation occurred rapidly in response to a brief UV-B treatment and no induction was observed in a uvr8 mutant over a broad range of UV wavelengths. In response to monochromatic light, maximal transcript accumulation occurred in wild-type plants at wavelengths 280-300 nm. HY5 transcript accumulation showed reciprocity between the fluence rate and duration of UV-B exposure, and on this basis conditions were chosen to generate an action spectrum for the UVR8 signaling pathway. Dose response curves were produced for a range of UV wavelengths using 20 min exposure to UV and harvesting tissue 2 h after the start of illumination. Experiments using mutants defective in sinapate ester and flavonoid biosynthesis indicated that the presence of UV-absorbing compounds did not affect the construction of an action spectrum under the conditions employed. The action spectrum for the induction of HY5 by the UVR8 pathway showed a main peak at 280 nm with a smaller peak at 300 nm. The data are discussed in relation to the proposed mechanisms of UV-B photoreception. PMID- 19558422 TI - Effectors of biotrophic fungi and oomycetes: pathogenicity factors and triggers of host resistance. AB - Many biotrophic fungal and oomycete pathogens share a common infection process involving the formation of haustoria, which penetrate host cell walls and form a close association with plant membranes. Recent studies have identified a class of pathogenicity effector proteins from these pathogens that is transferred into host cells from haustoria during infection. This insight stemmed from the identification of avirulence (Avr) proteins from these pathogens that are recognized by intracellular host resistance (R) proteins. Oomycete effectors contain a conserved translocation motif that directs their uptake into host cells independently of the pathogen, and is shared with the human malaria pathogen. Genome sequence information indicates that oomycetes may express several hundred such host-translocated effectors. Elucidating the transport mechanism of fungal and oomycete effectors and their roles in disease offers new opportunities to understand how these pathogens are able to manipulate host cells to establish a parasitic relationship and to develop new disease-control measures. PMID- 19558423 TI - Differential petiole growth in Arabidopsis thaliana: photocontrol and hormonal regulation. AB - Environmental challenges such as low light intensity induce differential growth driven upward leaf movement (hyponastic growth) in Arabidopsis thaliana. However, little is known about the physiological regulation of this response. Here, we studied how low light intensity is perceived and translated into a differential growth response in Arabidopsis. We used mutants defective in light, ethylene and auxin signaling, and in polar auxin transport, as well as chemical inhibitors, to analyze the mechanisms of low light intensity-induced differential growth. Our data indicate that photosynthesis-derived signals and blue light wavelengths affect petiole movements and that rapid induction of hyponasty by low light intensity involves functional cryptochromes 1 and 2, phytochrome-A and phytochrome-B photoreceptor proteins. The response is independent of ethylene signaling. Auxin and polar auxin transport, by contrast, play a role in low light intensity-induced differential petiole growth. We conclude that low light intensity-induced differential petiole growth requires blue light, auxin signaling and polar auxin transport and is, at least in part, genetically separate from well-characterized ethylene-induced differential growth. PMID- 19558424 TI - Large-scale parallel 454 sequencing reveals host ecological group specificity of arbuscular mycorrhizal fungi in a boreonemoral forest. AB - * Knowledge of the diversity of arbuscular mycorrhizal fungi (AMF) in natural ecosystems is a major bottleneck in mycorrhizal ecology. Here, we aimed to apply 454 sequencing--providing a new level of descriptive power--to assess the AMF diversity in a boreonemoral forest. * 454 sequencing reads of the small subunit ribosomal RNA (SSU rRNA) gene of Glomeromycota were assigned to sequence groups by blast searches against a custom-made annotated sequence database. * We detected 47 AMF taxa in the roots of 10 plant species in a 10 x 10 m plot, which is almost the same as the number of plant species in the whole studied forest. There was a significant difference between AMF communities in the roots of forest specialist plant species and in the roots of habitat generalist plant species. Forest plant species hosted 22 specialist AMF taxa, and the generalist plants shared all but one AMF taxon with forest plants, including globally distributed generalist fungi. These AMF taxa that have been globally recorded only in forest ecosystems were significantly over-represented in the roots of forest plant species. * Our findings suggest that partner specificity in AM symbiosis may occur at the level of ecological groups, rather than at the species level, of both plant and fungal partners. PMID- 19558425 TI - 5-HT 3 receptors mediate the time-dependent vagal afferent modulation of nociception during chronic food allergen-sensitized visceral hyperalgesia in rats. AB - Converging lines of evidence demonstrate a vagally mediated antinociceptive pathway in animals undergoing acute visceral insults, the contribution of this system to visceral pain following chronic noxious stimuli is unknown. 5-HT(3) receptor (5-HT(3)Rs) on spinal afferents are crucially involved in nociceptive processing, the role of 5-HT(3)Rs on vagal afferents is unclear. The aim of the present study was to determine the contribution of vagal afferents to visceral nociception in rats undergoing chronic luminal allergen stimulation and whether it involves vagal 5-HT(3)Rs. Sensitized rats received chicken egg albumin (EA, 1 mg mL(-1)) in drinking water for 2 weeks (day 1-14). Visceromotor response (VMR) to colorectal distension [colorectal distension (CRD), 60 mmHg] and the levels of mRNA encoding 5-HT(3)R (including 3A and 3B subunits) in the nodose ganglia (NG) were evaluated on day 2, 4, 8 and 15. Chronic EA challenge induced gradually increased visceral nociception, with a peak on day 15. Subdiaphragmatic vagotomy or functional deafferentation with capsaicin abolished this time-dependent manner, inducing hyperalgesia from day 2, lasting to day 15. Intraluminal infusion of a 5-HT(3)R antagonist (granisetron), whether alone or infused after local mucosa anaesthetic with 1% lidocaine, mimicked the effects of vagotomy. The mRNA levels for 5-HT(3B) or 5-HT(3A) subunit in the NG showed an opposite time course to that of visceral pain, which increased from day 2, then decreased gradually to levels lower than those of controls. Our results demonstrate a time dependent vagal afferent modulation of chronic allergen-sensitized visceral hyperalgesia, which may involve a 5-HT(3)R pathway. PMID- 19558426 TI - Increased mucosal nitric oxide production in ulcerative colitis is mediated in part by the enteroglial-derived S100B protein. AB - In the central nervous system glial-derived S100B protein has been associated with inflammation via nitric oxide (NO) production. As the role of enteroglial cells in inflammatory bowel disease has been poorly investigated in humans, we evaluated the association of S100B and NO production in ulcerative colitis (UC). S100B mRNA and protein expression, inducible NO synthase (iNOS) expression, and NO production were evaluated in rectal biopsies from 30 controls and 35 UC patients. To verify the correlation between S100B and NO production, biopsies were exposed to S100B, in the presence or absence of specific receptor for advanced glycation end-products (RAGE) blocking antibody, to measure iNOS expression and nitrite production. S100B and iNOS expression were evaluated after incubation of biopsies with lipopolysaccharides (LPS) + interferon-gamma (IFN gamma) in the presence of anti-RAGE or anti-S100B antibodies or budesonide. S100B mRNA and protein expression, iNOS expression and NO production were significantly higher in the rectal mucosa of patients compared to that of controls. Exogenous S100B induced a significant increase in both iNOS expression and NO production in controls and UC patients; this increase was inhibited by specific anti-RAGE blocking antibody. Incubation with LPS + IFN-gamma induced a significant increase in S100B mRNA and protein expression, together with increased iNOS expression and NO production. LPS + IFN-gamma-induced S100B up-regulation was not affected by budesonide, while iNOS expression and NO production were significantly inhibited by both specific anti-RAGE and anti-S100B blocking antibodies. Enteroglial derived S100B up-regulation in UC participates in NO production, involving RAGE in a steroid insensitive pathway. PMID- 19558427 TI - Electroacupuncture attenuates visceral hyperalgesia and inhibits the enhanced excitability of colon specific sensory neurons in a rat model of irritable bowel syndrome. AB - The causes of irritable bowel syndrome remain elusive and there are few effective treatments for pain in this syndrome. Electroacupunture (EA) is used extensively for treatment of various painful conditions including chronic visceral hyperalgesia (CVH). However, mechanism of its analgesic effect remains unknown. This study was designed to investigate effect of EA on colon specific dorsal root ganglion (DRG) neurons in rats with CVH. CVH was induced by intracolonic injection of acetic acid (AA) in 10-day-old rats. Electromyography and patch clamp recordings were performed at age of 8-10 weeks. Colon DRG neurons were labelled by injection of DiI into the colon wall. EA was given at ST36 in both hindlimbs. As adults, neonatal AA-injected rats displayed an increased sensitivity to colorectal distension (CRD) and an enhanced excitability of colon DRG neurons. EA treatment for 40 min significantly attenuated the nociceptive responses to CRD in these rats; this attenuation was reversed by pretreatment with naloxone. EA treatment for 40 min per day for 5 days produced a prolonged analgesic effect and normalized the enhanced excitability of colon DRG neurons. Furthermore, in vitro application of [D-Ala(2), N-MePhe(4), Gly(5)-Ol] enkephalin (DAMGO) suppressed the enhanced excitability of colon neurons from rats with CVH. These findings suggest that EA produced-visceral analgesia, which might be mediated in a large part by endogenous opioids pathways, is associated with reversal of the enhanced excitability of colon DRG neurons in rats with CVH. PMID- 19558428 TI - Study to compare the efficacy and safety of fluconazole cream with flutrimazole cream in the treatment of superficial mycosis: a multicentre, randomised, double blind, phase III trial. AB - Fluconazole, which is a drug of the azole family, is safely used in systemic treatment of oral and intravenous injection, but it is difficult to use fluconazole as a topical application because of its large molecular weight and strong hydrophilic property. This study is a multicentre, double-blind, randomised, non-inferiority study to compare the antifungal effect and safety of fluconazole cream 0.5% and 1% with flutrimazole cream 1% in superficial mycosis. A total of 162 subjects selected to participate in this study were equally divided into three groups and assigned to be given fluconazole cream 0.5%, fluconazole cream 1%, and flutrimazole cream 1% in the ratio of 1 : 1. The primary index of drug efficacy was determined by complete mycological cure in which no fungus was detected on KOH smear test 4 weeks after application of fluconazole. The secondary index of efficacy was defined as complete mycological cure 4 weeks after the application of fluconazole, improvement of clinical symptoms and overall effectiveness assessed by the research staff. According to this study, on comparing the efficacy of cure of superficial dermatomycosis after 4 weeks of application, both fluconazole 0.5% and fluconazole 1% cream were found to be equally effective and non-inferior to flutrimazole 1% cream. Given the effectiveness and safety of the drug, both fluconazole 0.5% and 1% cream might be said to be optimal concentration in the treatment of superficial dermatomycosis. PMID- 19558429 TI - Primary cutaneous cryptococcosis in a renal transplant recipient: case report. AB - We report a kidney transplant recipient with severe skin- and soft-tissue infection mimicking necrotising fasciitis. Patient failed to respond to empirical antibiotic therapy for presumed bacterial cellulitis. Culture of aspirate from the wound and tissue samples revealed Cryptococcus neoformans. No signs of systemic cryptococcal infection were found. After antifungal treatment and surgical intervention, complete healing was achieved. Clinical and microbiological characteristics of this patient are discussed. Our case indicates that primary cutaneous cryptococcosis must be included in the differential diagnosis of severe cellulitis in solid organ transplant recipients not responding to broad-spectrum antibiotic regimens. In our case, prompt diagnosis and treatment could dramatically modify the outcome. PMID- 19558430 TI - Widespread erosive tinea corporis by Arthroderma benhamiae in a renal transplant recipient: case report. AB - Superficial fungal infections are expected to be more prevalent in renal transplant recipients because of graft-preserving immunosuppressive therapy. Here we report the case of a 45-year-old patient with widespread erosive tinea corporis transmitted by domestic animals. Sequencing of the internal transcribed spacer region identified Arthroderma benhamiae (teleomorph of Trichophyton mentagrophytes) in the patient, her husband and her domestic animals. A combination therapy with systemic terbinafine hydrochloride and topically applied ciclopiroxolamine was successful. PMID- 19558431 TI - Tinea incognito due to Trichophyton mentagrophytes. AB - Tinea incognito is a dermatophytosis of atypical clinical character, usually misdiagnosed and treated with corticosteroids. We report a case of tinea faciei modified by high potency topical corticosteroids in a 54-year-old woman. Deep, intense inflammatory plaque with boggy, pustular surface located on the right cheek was found. Direct microscopy and culture confirmed dermatophytosis and led to the identification of Trichophyton mentagrophytes var. mentagrophytes. Complete resolution occurred after treatment with oral terbinafine. PMID- 19558433 TI - Non-alcoholic fatty liver disease and adipocytokines. PMID- 19558434 TI - Identification of five novel 14-3-3 isoforms interacting with the GPIb-IX complex in platelets. AB - BACKGROUND: Binding of von Willebrand factor to the platelet glycoprotein (GP)Ib IX complex initiates a signaling cascade leading to integrin alpha(IIb)beta(3) activation, a key process in hemostasis and thrombosis. Interaction of 14-3-3zeta with the intracytoplasmic domain of GPIb appears to be a major effector of this activation pathway. OBJECTIVE: The aim of our study was to determine whether other members of the 14-3-3 family bind to GPIb-IX. RESULTS: In this study, western blot analyses showed that platelets also contain the 14-3-3beta, 14-3 3gamma, 14-3-3epsilon, 14-3-3eta and 14-3-3theta isoforms, but lack 14-3-3sigma. Coimmunoprecipitation studies in platelets and CHO transfectants demonstrated that all six 14-3-3 isoforms expressed in platelets, including, as previously reported, 14-3-3zeta, bind to GPIb-IX. In addition, their interaction was found to critically require the same GPIbalpha domains (580-590 and 605-610) already identified as essential for 14-3-3zeta binding, in agreement with the conservation of the sequence of the I-helix among these different isoforms. Pull down experiments indicated that all six 14-3-3 isoforms present in platelets bind to GPIbbeta. In contrast, deletion or mutation of the GPIbbeta intracytoplasmic tail did not affect the interaction of GPIb-IX with the 14-3-3 isoforms, questioning the importance of this domain. CONCLUSIONS: Our study suggests that, to inhibit GPIb-induced integrin alpha(IIb)beta(3) activation, a more appropriate strategy than inhibiting individual 14-3-3 isoforms would be to target the 14-3-3 binding motif on GPIb or, alternatively, the conserved 14-3-3 I-helix. PMID- 19558432 TI - Dissecting the essentiality of the bifunctional trypanothione synthetase-amidase in Trypanosoma brucei using chemical and genetic methods. AB - The bifunctional trypanothione synthetase-amidase (TRYS) comprises two structurally distinct catalytic domains for synthesis and hydrolysis of trypanothione (N(1),N(8)-bis(glutathionyl)spermidine). This unique dithiol plays a pivotal role in thiol-redox homeostasis and in defence against chemical and oxidative stress in trypanosomatids. A tetracycline-dependent conditional double knockout of TRYS (cDKO) was generated in bloodstream Trypanosoma brucei. Culture of cDKO parasites without tetracycline induction resulted in loss of trypanothione and accumulation of glutathione, followed by growth inhibition and cell lysis after 6 days. In the absence of inducer, cDKO cells were unable to infect mice, confirming that this enzyme is essential for virulence in vivo as well as in vitro. To establish whether both enzymatic functions were essential, an amidase-dead mutant cDKO line was generated. In the presence of inducer, this line showed decreased growth in vitro and decreased virulence in vivo, indicating that the amidase function is not absolutely required for viability. The druggability of TRYS was assessed using a potent small molecule inhibitor developed in our laboratory. Growth inhibition correlated in rank order cDKO, single KO, wild-type and overexpressing lines and produced the predicted biochemical phenotype. The synthetase function of TRYS is thus unequivocally validated as a drug target by both chemical and genetic methods. PMID- 19558435 TI - Long-term thrombin inhibition promotes cancer cell extravasation in a mouse model of experimental metastasis. PMID- 19558436 TI - The paradox of heparin-induced thrombocytopenia. PMID- 19558438 TI - Seeing my world in a million little pieces: narcissism, self-construal, and cognitive-perceptual style. AB - In 4 studies we examine the association between narcissism, self-construal, and cognitive-perceptual style, hypothesizing that high self-focus in combination with low other-focus (i.e., social atomization) is related to an analytic cognitive-perceptual style. Participants completed the Narcissistic Personality Inventory, the Self-Construal Scale, and measures of cognitive-perceptual style such as the Analysis-Holism Scale, the Embedded Figures Test, a visual illusion test, and a measure of the representativeness heuristic. We found evidence for a decontextualized cognitive-perceptual style in socially atomized participants, which included those high in narcissism and also those who had a combination of high independent and low interdependent self-construal. A meta-analytic integration of our findings found that narcissism was positively related to independent and negatively related to interdependent self-construal, and mediation analyses found some evidence that the relationship between self construal and cognitive-perceptual style is partially mediated by narcissism. PMID- 19558437 TI - Reciprocal relations among self-efficacy beliefs and prosociality across time. AB - The present study examined the longitudinal relations between individuals' prosociality and their self-efficacy beliefs in regard to emotional regulation and responding empathically to others' needs. The participants were 244 females and 222 males with a mean age of 17 years (SD=1.5) at T1, 19 years (SD=1.4) at T2, and 21 years (SD=1.6) at T3. The findings corroborated the posited paths of relations assigning empathic self-efficacy a major role in predicting the level of individuals' prosociality. Empathic self-efficacy beliefs mediated the relations of regulative emotional self-efficacy beliefs to prosocial tendencies such as caring, sharing, helping, and empathic concern toward others. The posited conceptual model accounted for a significant portion of variance in prosociality and has implications for interventions designed to promote and sustain prosociality. PMID- 19558439 TI - What constitutes a good life? Cultural differences in the role of positive and negative affect in subjective well-being. AB - East Asians and Asian Americans report lower levels of subjective well-being than Europeans and European Americans. Three studies found support for the hypothesis that such differences may be due to the psychological meanings Eastern and Western cultures attach to positive and negative affect. Study 1 demonstrated that the desire to repeat a recent vacation was significantly predicted by recalled positive affect-but not recalled negative affect-for European Americans, whereas Asian Americans considered both positive and negative affect. Study 2 replicated this effect in judging satisfaction with a personal friendship. Study 3 linked changes in European Americans' life satisfaction to everyday positive events caused by the self (vs. others) and changes in Japanese life satisfaction to everyday negative events caused by others (vs. the self). Positive affect appears particularly meaningful for European Americans and negative affect for Asian Americans and Japanese when judging a satisfying vacation, friendship, or life. PMID- 19558440 TI - Aggression toward gay men as gender role enforcement: effects of male role norms, sexual prejudice, and masculine gender role stress. AB - This study examined sexual prejudice and masculine gender role stress as mediators of the relations between male gender norms and anger and aggression toward gay men. Participants were 150 self-identified heterosexual men who completed measures of adherence to male gender role norms, sexual prejudice, masculine gender role stress, and state anger. Participants then viewed a video depicting intimate relationship behavior between 2 gay men, reported state anger a second time, and competed in a laboratory aggression task against either a heterosexual or a gay male. Results indicated that adherence to the antifemininity norm exerted an indirect effect, primarily through sexual prejudice, on increases in anger. Adherence to the status and antifemininity norms exerted indirect effects, also through sexual prejudice, on physical aggression toward the gay, but not the heterosexual, male. Findings provide the first multivariate evidence for determinants of aggression toward gay men motivated by gender role enforcement. PMID- 19558441 TI - Appraisal underpinnings of affective chronometry: the role of appraisals in emotion habituation. AB - In light of the lack of studies examining the cognitive components of affective chronometry, this research examined the appraisals associated with emotion habituation, using anger as the emotion of focus. Anger and its appraisals were assessed repeatedly over a day in the participants' naturalistic contexts. The trajectory of decline in anger over time after its first appearance was found to be a function of chronic coping styles. More importantly, the trajectories of anger-related appraisals generally corresponded to that of anger and were also moderated by coping styles in ways consistent with the moderating effects of coping styles on anger. Implications of these results for affective chronometry research and for appraisal research are discussed. PMID- 19558442 TI - Metatraits of the Big Five differentially predict engagement and restraint of behavior. AB - Although initially believed to contain orthogonal dimensions, the Big Five personality taxonomy appears to have a replicable higher-order structure, with the metatrait of Plasticity reflecting the shared variance between Extraversion and Openness/Intellect, and the metatrait of Stability reflecting the shared variance among Neuroticism, Agreeableness, and Conscientiousness. These higher order traits have been theorized to relate to individual differences in the functioning of the dopamine and serotonin systems, respectively. As dopamine is associated with exploration and incentive-related action, and serotonin with satiety and constraint, this neuropharmacological trait theory has behavioral implications, which we tested in 307 adults by examining the association of a large number of behavioral acts with multi-informant reports of the metatraits. The frequencies of acts were consistently positively correlated with Plasticity and negatively correlated with Stability. At the broadest level of description, variation in human personality appears to reflect engagement and restraint of behavior. PMID- 19558443 TI - The influence of work personality on job satisfaction: incremental validity and mediation effects. AB - Drawing from recent developments regarding the contextual nature of personality (e.g., D. Wood & B. W. Roberts, 2006), we conducted 2 studies (1 cross-sectional and 1 longitudinal over 1 year) to examine the validity of work personality in predicting job satisfaction and its mediation of the effect of global personality on job satisfaction. Study 1 showed that (a) individuals vary systematically in their personality between roles- they were significantly more conscientious and open to experience and less extraverted at work compared to at home; (b) work personality was a better predictor of job satisfaction than both global personality and home personality; and (c) work personality demonstrated incremental validity above and beyond the other two personality measures. Study 2 further showed that each of the work personality dimensions fully mediated the association between its corresponding global personality trait and job satisfaction. Evidence for the discriminant validity of the findings is also presented. PMID- 19558444 TI - The hierarchical structure of well-being. AB - Theories of hedonic, eudaimonic, and social well-being provide 3 extensively studied models for explaining flourishing mental health. Few studies have examined whether these models can be integrated into a comprehensive structure of well-being. The present study builds upon previous theoretical and empirical work to determine the complex relationships among these 3 models of well-being. Confirmatory factor analysis techniques were used to test a series of models in order to (a) confirm the proposed latent structures of hedonic, eudaimonic, and social well-being and (b) examine whether these models could be successfully integrated into a hierarchical structure of well-being. In 2 large samples, results supported the proposed latent structures of hedonic, eudaimonic, and social well-being and indicated that the various components of well-being could be represented most parsimoniously with 3 oblique second-order constructs of hedonic, eudaimonic, and social well-being. PMID- 19558445 TI - The pathoplastic relationship between interpersonal problems and fear of failure. AB - This study advances the understanding of fear of failure (FF), a dispositional avoidance-oriented achievement motive, by employing interpersonal classification techniques to groups of individuals who fear failure in order to examine the pathoplastic relations between FF and interpersonal problems. Shame-based FF is thought to be related to the self-regulation strategies of appeasement and aggression, and these strategies are hypothesized to be associated with the interpersonal problems of Nonassertiveness and Vindictiveness, respectively. Using 2 independent samples (ns=60 and 38) reporting high FF, interpersonal profiles were examined for the groups in their entirety and for cluster solutions within the larger samples. Results demonstrated that individuals high in FF exhibited 1 of 2 prototypical interpersonal profiles associated with Domineering/Vindictive or Nonassertive/Exploitable problems that correspond with the hypothesized aggression and appeasement regulation strategies. Consistent with the concept of pathoplasticity, these differences were not better accounted for by demographic, affective, motivational, personality, or attachment style characteristics of the samples. PMID- 19558446 TI - Comparing two alternative measures of general personality in the assessment of psychopathy: a test of the NEO PI-R and the MPQ. AB - This study examined the interrelations between two measures of personality, the Revised NEO Personality Inventory (NEO PI-R; P. T. Costa & R. R. McCrae, 1992) and the Multidimensional Personality Questionnaire (MPQ; Tellegen & Waller, 2008), and their relations with psychopathy in a sample of undergraduates. Results revealed good convergence between conceptually related personality traits; however, the NEO PI-R facets accounted for more variance in the MPQ subscales (mean R(2)=.49) than did MPQ subscales in NEO PI-R facets (mean R(2)=.35). Both accounted for substantial proportions of variance in psychopathy scores, although the NEO PI-R accounted for larger proportions and manifested greater incremental validity when using the broader domains of each measure; the differences decreased when the narrower facets/subscales were used. The results suggest that, although both measures assess psychopathy-related traits, the NEO PI-R provides a more complete description because of its assessment of interpersonal antagonism and the central role of this construct in psychopathy. PMID- 19558447 TI - What leads to romantic attraction: similarity, reciprocity, security, or beauty? Evidence from a speed-dating study. AB - Years of attraction research have established several "principles" of attraction with robust evidence. However, a major limitation of previous attraction studies is that they have almost exclusively relied on well-controlled experiments, which are often criticized for lacking ecological validity. The current research was designed to examine initial attraction in a real-life setting-speed-dating. Social Relations Model analyses demonstrated that initial attraction was a function of the actor, the partner, and the unique dyadic relationship between these two. Meta-analyses showed intriguing sex differences and similarities. Self characteristics better predicted women's attraction than they did for men, whereas partner characteristics predicted men's attraction far better than they did for women. The strongest predictor of attraction for both sexes was partners' physical attractiveness. Finally, there was some support for the reciprocity principle but no evidence for the similarity principle. PMID- 19558448 TI - The political is personal: narrating 9/11 and psychological well-being. AB - Making meaning out of negative experiences is one of the primary psychological challenges in the wake of adversity. Much of the empirical attention that psychologists have paid to meaning making has focused on personal hardships, but national tragedies similarly pose a challenge to meaning making. In the present study, which is grounded in the theoretical tradition of the narrative study of lives, a nationally representative sample of 395 adults wrote accounts about the 9/11 terrorist attacks approximately 2 months after 9/11. Accounts were coded for 3 narrative themes: closure, redemption, and contamination. Psychological well being was significantly related to accounts that were high in closure and national redemption and, among those more directly exposed to the attacks, accounts high in redemptive imagery. Psychological distress was significantly related to accounts that were low in closure and high in themes of personal contamination. Understanding the narrative styles that characterize personal accounts of political events has important ramifications for the study of the socially embedded individual. PMID- 19558449 TI - Manganese induces sustained Ser40 phosphorylation and activation of tyrosine hydroxylase in PC12 cells. AB - Manganese (Mn2+) is an essential metal involved in normal functioning of a range of physiological processes. However,occupational overexposure to Mn2+ causes neurotoxicity. The dopaminergic system is a particular target for Mn2+ neurotoxicity.Tyrosine hydroxylase (TH) is the rate limiting enzyme for dopamine synthesis and is regulated acutely by phosphorylation at Ser40 and chronically by protein synthesis. In this study we used pheochromocytoma 12 cells to investigate the effects of Mn2+ exposure on the phosphorylation and activity of TH. Mn2+ treatment for 24 h caused a sustained increase in Ser40 phosphorylation and TH activity at a concentration of 100 lM, without altering the level of TH protein orPC12 cell viability. Inhibition of protein kinase A and protein kinase C and protein kinases known to be involved in sustained phosphorylation of TH in response to other stimuli didnot block the effects of Mn2+ on Ser40 phosphorylation.A substantial increase in H2O2 production occurred in response to 100 lM Mn2+. The antioxidant Trolox completely inhibited H2O2 production but did not block TH phosphorylation at Ser40, indicating that oxidative stress was not involved. Sustained TH phosphorylation at Ser40 and the consequent activation of TH both occurred at low concentrations of Mn2+ and this provides a potential new mechanism for Mn2+-induced neuronal action that does not involve H2O2-mediated cell death. PMID- 19558450 TI - GABA action in immature neocortical neurons directly depends on the availability of ketone bodies. AB - In the early postnatal period, energy metabolism in the suckling rodent brain relies to a large extent on metabolic pathways alternate to glucose such as the utilization of ketone bodies (KBs). However, how KBs affect neuronal excitability is not known. Using recordings of single NMDA and GABA-activated channels in neocortical pyramidal cells we studied the effects of KBs on the resting membrane potential (E(m)) and reversal potential of GABA-induced anionic currents (E(GABA)), respectively. We show that during postnatal development (P3-P19) if neocortical brain slices are adequately supplied with KBs, E(m) and E(GABA) are both maintained at negative levels of about -83 and -80 mV, respectively. Conversely, a KB deficiency causes a significant depolarization of both E(m) (>5 mV) and E(GABA) (>15 mV). The KB-mediated shift in E(GABA) is largely determined by the interaction of the NKCC1 cotransporter and Cl(-)/HCO3 transporter(s). Therefore, by inducing a hyperpolarizing shift in E(m) and modulating GABA signaling mode, KBs can efficiently control the excitability of neonatal cortical neurons. PMID- 19558452 TI - The SIRT1 activator resveratrol protects SK-N-BE cells from oxidative stress and against toxicity caused by alpha-synuclein or amyloid-beta (1-42) peptide. AB - Human sirtuins are a family of seven conserved proteins (SIRT1-7). The most investigated is the silent mating type information regulation-2 homolog (SIRT1, NM_012238), which was associated with neuroprotection in models of polyglutamine toxicity or Alzheimer's disease (AD) and whose activation by the phytocompound resveratrol (RES) has been described. We have examined the neuroprotective role of RES in a cellular model of oxidative stress, a common feature of neurodegeneration. RES prevented toxicity triggered by hydrogen peroxide or 6 hydroxydopamine (6-OHDA). This action was likely mediated by SIRT1 activation, as the protection was lost in the presence of the SIRT1 inhibitor sirtinol and when SIRT1 expression was down-regulated by siRNA approach. RES was also able to protect SK-N-BE from the toxicity arising from two aggregation-prone proteins, the AD-involved amyloid-beta (1-42) peptide (Abeta42) and the familiar Parkinson's disease linked alpha-synuclein(A30P) [alpha-syn(A30P)]. Alpha syn(A30P) toxicity was restored by sirtinol addition, while a partial RES protective effect against Abeta42 was found even in presence of sirtinol, thus suggesting a direct RES effect on Abeta42 fibrils. We conclude that SIRT1 activation by RES can prevent in our neuroblastoma model the deleterious effects triggered by oxidative stress or alpha-syn(A30P) aggregation, while RES displayed a SIRT1-independent protective action against Abeta42. PMID- 19558451 TI - Subcellular compartment-specific molecular diversity of pre- and post-synaptic GABA-activated GIRK channels in Purkinje cells. AB - Activation of G protein-gated inwardly-rectifying K(+) (GIRK or Kir3) channels by metabotropic gamma-aminobutyric acid (B) (GABA(B)) receptors is an essential signalling pathway controlling neuronal excitability and synaptic transmission in the brain. To investigate the relationship between GIRK channel subunits and GABA(B) receptors in cerebellar Purkinje cells at post- and pre-synaptic sites, we used biochemical, functional and immunohistochemical techniques. Co immunoprecipitation analysis demonstrated that GIRK subunits are co-assembled with GABA(B) receptors in the cerebellum. Immunoelectron microscopy showed that the subunit composition of GIRK channels in Purkinje cell spines is compartment dependent. Thus, at extrasynaptic sites GIRK channels are formed by GIRK1/GIRK2/GIRK3, post-synaptic densities contain GIRK2/GIRK3 and dendritic shafts contain GIRK1/GIRK3. The post-synaptic association of GIRK subunits with GABA(B) receptors in Purkinje cells is supported by the subcellular regulation of the ion channel and the receptor in mutant mice. At pre-synaptic sites, GIRK channels localized to parallel fibre terminals are formed by GIRK1/GIRK2/GIRK3 and co-localize with GABA(B) receptors. Consistent with this morphological evidence we demonstrate their functional interaction at axon terminals in the cerebellum by showing that GIRK channels play a role in the inhibition of glutamate release by GABA(B) receptors. The association of GIRK channels and GABA(B) receptors with excitatory synapses at both post- and pre-synaptic sites indicates their intimate involvement in the modulation of glutamatergic neurotransmission in the cerebellum. PMID- 19558453 TI - A highly conserved tryptophan residue in the fourth transmembrane domain of the A adenosine receptor is essential for ligand binding but not receptor homodimerization. AB - Dimerization between G protein-coupled receptors (GPCRs) is a clearly established phenomenon. However, limited information is currently available on the interface essential for this process. Based on structural comparisons and sequence homology between rhodopsin and A(1) adenosine receptor (A(1)R), we initially hypothesized that four residues in transmembrane (TM) 4 and TM5 are involved in A(1)R homodimerization. Accordingly, these residues were substituted with Ala by site directed mutagenesis. Interestingly, the mutant protein displayed no significant decrease in homodimer formation compared with wild-type A(1)R, as evident from coimmunoprecipitation and BRET(2) analyses (improved bioluminescence resonance energy transfer system offered by Perkin-Elmer Life Sciences), but lost ligand binding activity almost completely. Further studies disclosed that this effect was derived from the mutation of one particular residue, Trp132, which is highly conserved among many GPCRs. Confocal immunofluorescence and cell-surface biotinylation studies revealed that the mutant receptors localized normally at transfected cell membranes, signifying that loss of ligand binding was not because of defective cellular trafficking. Molecular modeling of the A(1)R-ligand complex disclosed that Trp132 interacted with several residues located in TM3 and TM5 that stabilized agonist binding. Thus, loss of interactions of Trp with these residues may, in turn, disrupt binding to agonists. Our study provides strong evidence of the essential role of the highly conserved Trp132 in TM4 of adenosine receptors. PMID- 19558454 TI - Alpha A-crystallin and alpha B-crystallin, newly identified interaction proteins of protease-activated receptor-2, rescue astrocytes from C2-ceramide- and staurosporine-induced cell death. AB - Protease-activated receptor-2 (PAR-2) is a G protein-coupled receptor activated by trypsin and other trypsin-like serine proteases. The widely expressed PAR-2 is involved in inflammation response but the physiological/pathological roles of PAR 2 in the nervous system are still uncertain. In the present study, we report novel PAR-2 interaction proteins, alphaA-crystallin and alphaB-crystallin. These 20 kDa proteins have been implicated in neurodegenerative diseases like Alexander's disease, Creutzfeldt-Jacob disease, Alzheimer's disease, and Parkinson's disease. Results from yeast two-hybrid assay using the cytoplasmic C tail of PAR-2 as bait suggested that alphaA-crystallin interacts with PAR-2. We further demonstrate the in vitro and cellular in vivo interaction of C-tail of PAR-2 as well as of full-length PAR-2 with alphaA(alphaB)-crystallins. We use pull-down, co-immunoprecipitation, and co-localization assays. Analysis of alphaA crystallin deletion mutants showed that amino acids 120-130 and 136-154 of alphaA crystallin are required for the interaction with PAR-2. Co-immunoprecipitation experiments ruled out an interaction of alphaA(alphaB)-crystallins with PAR-1, PAR-3, and PAR-4. This demonstrates that alphaA(alphaB)-crystallins are PAR-2 specific interaction proteins. Moreover, we investigated the functional role of PAR-2 and alpha-crystallins in astrocytes. Evidence is presented to show that PAR 2 activation and increased expression of alpha-crystallins reduced C2-ceramide- and staurosporine-induced cell death in astrocytes. Thus, both PAR-2 and alpha crystallins are involved in cytoprotection in astrocytes. PMID- 19558455 TI - A protein-protein interaction of stress-responsive myosin VI endowed to inhibit neural progenitor self-replication with RNA binding protein, TLS, in murine hippocampus. AB - We have shown preferential expression of both mRNA and corresponding protein for myosin VI (Myo6) in the murine hippocampus within 24 h after the extreme traumatic experience, water-immersion restraint stress (WIRS), prior to a drastic decrease in neural progenitor proliferation in the dentate gyrus. Myosin (Myo6) protein levels were significantly increased in hippocampus within 24 h after flashback experience in mice previously exposed to WIRS. Myo6 protein was ubiquitously distributed in discrete mouse brain regions with exceptionally high expression in olfactory bulb, whereas Myo6 protein was expressed in cultured rat astroglia and neurons, in addition to Myo6 mRNA expression by cultured neural progenitors. In mouse embryonal carcinoma P19 cells endowed to proliferate and differentiate, Myo6 protein was expressed in line with astroglial marker protein expression. Transient over-expression of Myo6 induced a significant decrease in the size of clustered aggregates as an index of self-replication in P19 cells. Immunoprecipitation analysis revealed the interaction between Myo6 and the RNA binding protein, translocated in liposarcoma (TLS), while TLS was predominantly expressed by neurons in the cortex, striatum, cerebellum, and hippocampus. These results suggest that Myo6 may play a pivotal role in the mechanism underlying the suppressed adult neurogenesis after traumatic stress in association with TLS. PMID- 19558457 TI - Paraplegia in a Bornean orangutan (Pongo pygmaeus pygmaeus) due to multiple myeloma. AB - BACKGROUND: A 38-year-old male Bornean orangutan (Pongo pygmaeus pygmaeus) developed progressive hind leg paresis. A computed tomography scan of the vertebral column revealed soft tissue type densities within vertebral bones. METHODS AND RESULTS: At necropsy infiltrating tumor masses were found in the vertebral bodies, protruding into the spinal canal and compressing the spinal cord. Microscopically neoplastic plasma cells infiltrated the vertebral bodies and adjacent soft tissues. Immunohistochemically, tumor cells tested positive for B cell markers (CD38, CD79alpha), kappa, and lambda light chains, while vimentin, GFAP, S100, and CD138 were not expressed. The tumor was classified as multiple myeloma on the basis of radiographic, pathological, and immunohistochemical findings. CONCLUSIONS: This first systematic case description on multiple myeloma in a non-human primate revealed many similarities with the disease in humans and the immunohistochemical tools proved suitable for their use in the orangutan. PMID- 19558456 TI - The muscarinic M1 receptor activates Nrf2 through a signaling cascade that involves protein kinase C and inhibition of GSK-3beta: connecting neurotransmission with neuroprotection. AB - In this study, we provide evidence that the muscarinic M1 receptor targets NF-E2 related factor-2 (Nrf2), a transcription factor that regulates the expression of genes containing antioxidant response elements (AREs) in their promoters and that collectively constitute the phase II antioxidant response. In hippocampal primary and cerebellar granule neuron cultures expressing endogenous M1 receptor, carbachol increased the levels of a prototypical phase II antioxidant enzyme, heme oxygenase-1. Moreover, in a heterologous system, based on lentiviral expression of M1 receptor in PC12 pheochromocytoma cells, we found that M1 increased total and nuclear Nrf2 protein levels and heme oxygenase-1 messenger RNA and protein levels. Luciferase reporter constructs for AREs and the use of two inhibitors of protein kinase C (PKC), chelerythrine and 2-aminoethyl diphenylborinate, or transfection with relevant expression vectors allowed us to identify Galphaq, phospholipase C-beta and the classical PKC-gamma isoenzyme, as responsible for the regulation of Nrf2. A PKC-insensitive Nrf2S40A single-point mutant partially channeled M1 signaling to AREs, therefore suggesting the participation of additional intermediates. Inhibition of glycogen synthase kinase 3beta (GSK-3beta) augmented M1-dependent activation of AREs while a PKC insensitive mutant of GSK-3beta (GSK-3beta-Delta9) blocked this effect and prevented M1-induced accumulation of Nrf2 in the nucleus. Our results demonstrate a previously unidentified role of the Galphaq/phospholipase C-beta/PKC/GSK-3beta axis in regulation of Nrf2 by M1. Such role provides additional conceptual support for the use of cholinemimetics in the treatment of pathologies that, like Alzheimer's disease, require a reinforcement of the cell antioxidant capacity. PMID- 19558458 TI - The significance of neighbours: views and experiences of people with intellectual disability on neighbouring. AB - BACKGROUND: People with intellectual disability (ID) who live in regular neighbourhoods have experiences with their neighbours, which are important to understand when studying social integration. METHOD: This study describes and analyses the opinions on, and experiences with, neighbour relationships of 39 people with ID living in neighbourhood housing facilities. RESULTS: We found that, while the views of people with ID on 'good neighbouring' were consistent with 'neighbouring' described in sociological literature, their experiences may be influenced by an organisational context, the tendency to formalise relationships and apprehension towards meeting unfamiliar people. CONCLUSION: Understanding influential factors to neighbouring for people with ID may shed light on the processes involved in social integration of people with ID at a neighbourhood level. This paper contributes to understanding the opinions of people with ID on satisfactory neighbourhood relationships, and explores opportunities to improve them. PMID- 19558459 TI - Correlation of visual function with health-related quality of life in glaucoma patients. AB - OBJECTIVES: The purpose of this study was to assess visual impairment and quality of life (QOL), the relationship between visual function and health-related and vision-related QOL, and to identify factors significantly impacting QOL for glaucoma patients. METHODS: One-on-one interviews of 280 glaucoma patients at a regional hospital in Taiwan from 1 April 2005 to 30 September 2005. Data collected included demographics, clinical examinations and treatment status. QOL data collected utilized the Chinese (Taiwanese) version of the Medical Outcomes Study Short-Form 36 Health Survey (SF-36) and the National Eye Institute Visual Function-25 (NEIVFQ-25). RESULTS: The majority of study participants had open angle glaucoma (54.3%) and were classified as moderate stage (33.6%). The higher the health-related QOL scores for both questionnaires, the fewer difficulties experienced by the patients. Moderate correlation was found between the SF-36 and NEIVFQ-25 subscales and visual function. Significant predicting factors for the SF-36 were gender, age, employment status, presence of systemic disease, better eye visual acuity and better-eye mean defect. For the NEIVFQ-25, these were general vision, near and distance activities, colour vision and peripheral vision scores. CONCLUSIONS: A combination of demographic data, clinical examination and QOL questionnaires may help clinicians better ascertain the impact of disease severity on glaucoma patients' QOL. This information may help in patient education, treatment compliance and selection of treatment options. PMID- 19558460 TI - Acne's relationship with psychiatric and psychological morbidity: results of a school-based cohort study of adolescents. AB - BACKGROUND: There is a considerable number of studies linking acne with psychological and psychiatric morbidities, although this literature is not entirely consistent and is largely cross-sectional in methodology. OBJECTIVE: This study aims to establish the relationship of acne and psychological and psychiatric morbidity in adolescents in a community setting and, via a longitudinal methodology, provide evidence for causality in the relationship. METHODS: The study was a 12-month cohort study. Two hundred and forty-four students in Years 8, 9 and 11 (ages 14-17) at four Australian high schools were assessed at baseline 6 months and 12 months. Presence and severity of acne were assessed, along with a number of psychological and psychiatric morbidities and personality traits (depression, anxiety, overall psychiatric morbidity, self consciousness, neuroticism and introversion/extraversion) and other demographic variables. RESULTS: Of the 244 participating students, 209 (86%) completed all three rounds of data collection. A further 26 (11%) completed two rounds. The study failed to demonstrate an association of the presence of acne or of acne severity with the examined measures of psychological and psychiatric morbidity, and no evidence for an effect of acne in their causation. CONCLUSION: The relationship of acne and psychological morbidities found in previous health care settings was not found in this community sample. This may be due to differences between community and clinical acne populations. Other possible reasons for this finding are attenuation of psychological morbidity in subjects in this study by successful acne treatment, and the role of personality traits in the complex relationship between acne and psychological morbidities. It is suggested that this relationship would be best investigated by means of longer-term cohort studies enlisting subjects at an early age, prior to the onset of acne. PMID- 19558462 TI - A 10-year review of cutaneous melanomas in Korea: clinicopathologic features and changes by multidisciplinary team approach. PMID- 19558463 TI - The interplay of lipopolysaccharide-binding protein and cytokines in periodontal health and disease. AB - AIM: Periodontal pathogenesis is characterized by Gram-negative bacteria activation of series of pro- and anti-inflammatory cytokines from host cells through the pathway of lipopolysaccharide (LPS), LPS-binding protein (LBP) and CD14. The present study investigated the expression profiles of interleukin (IL) 1beta and IL-10 in periodontal health and disease, and examined the effects of Escherichia coli LPS and LBP interaction on the expression of IL-1beta and IL-10 by human gingival fibroblasts (HGF). MATERIAL AND METHODS: Gingival biopsies were collected from 44 subjects with chronic periodontitis and 15 periodontally healthy subjects. The expression of IL-1beta and IL-10 was detected by immunohistochemistry. The mRNA expression of IL-1beta and IL-10 in HGF was detected by RT-PCR with or without recombinant human LBP (rhLBP), while the peptides were analysed by an enzyme-linked immunosorbent assay. RESULTS: IL-1beta was detected in both oral sulcular epithelia of healthy controls and periodontal pocket epithelia of patients. IL-10 was mainly expressed in the intercellular spaces of connective tissues. IL-1beta displayed a reverse pattern of expression levels with reference to IL-10, and a negative correlation existed between LBP and the ratio of IL-1beta/IL-10. rhLBP suppressed E. coli LPS-induced IL-1beta expression by HGF. CONCLUSION: An appropriate interplay of LBP and cytokines may have a beneficial effect on innate host defence, thereby contributing to periodontal homeostasis. PMID- 19558465 TI - Effects of selected pectinolytic bacterial strains on water-retting of hemp and fibre properties. AB - AIMS: To study the effect of selected bacterial strains on hemp water-retting and properties of retted fibre. METHODS AND RESULTS: The trials were performed in laboratory tanks. The traditional water-retting process, without inoculum addition, was compared to a process modified by inoculating water tanks with two selected pectinolytic bacteria: the anaerobic strain Clostridium sp. L1/6 and the aerobic strain Bacillus sp. ROO40B. Six different incubation times were compared. Half the fibre obtained from each tank was combed. Micromorphological analyses were performed by scanning electron microscopy on uncombed and combed fibres. Moreover, organoleptic and chemical analyses of uncombed fibres were performed. CONCLUSIONS: The inoculum, besides speeding up the process, significantly improved the fibre quality. The fibre was not damaged by mechanical hackling, thanks to the good retting level obtained by the addition of selected strains, differently to what happened with the traditionally retted fibre. The best fibre quality was obtained after 3-4 days of retting with the addition of the bacterial inoculum. SIGNIFICANCE AND IMPACT OF THE STUDY: Retting is the major limitation to an efficient production of high-quality hemp fibres. The water-retting process and fibre quality were substantially improved by simultaneously inoculating water tanks with two selected pectinolytic strains. PMID- 19558466 TI - Comparison of five commercial DNA extraction kits for the recovery of Yersinia pestis DNA from bacterial suspensions and spiked environmental samples. AB - AIM: To evaluate commercial DNA extraction kits for their ability to isolate DNA from Yersinia pestis suspensions and spiked environmental samples. METHODS AND RESULTS: Five commercially available DNA extraction kits were evaluated: the ChargeSwitch gDNA Mini Bacteria Kit, the IT 1-2-3 Sample DNA Purification Kit, the MasterPure Complete DNA and RNA Purification Kit, the QIAamp DNA Blood Mini Kit and the UltraClean Microbial DNA Isolation Kit. The extraction methods were performed upon six Y. pestis strains and spiked environmental specimens, including three swab types and one powder type. Taqman real-time PCR analysis revealed that the use of the MasterPure kit resulted in DNA with the most consistently positive results and the lowest limit of detection from Y. pestis suspensions and spiked environmental samples. CONCLUSION: Comparative evaluations of the five commercial DNA extraction methods indicated that the MasterPure kit was superior for the isolation of PCR-amplifiable DNA from Y. pestis suspensions and spiked environmental samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study can assist diagnostic laboratories with selecting the best extraction method for processing environmental specimens for subsequent detection of Y. pestis by real-time PCR. PMID- 19558467 TI - Screening the Fusarium graminearum inhibitory mutant strain from Bacillus subtilis by atmospheric-pressure plasma jet. AB - AIMS: The aim of this study was to improve the antagonistic activity of Bacillus subtilis JA towards Fusarium graminearum by screening high-yielding mutant using the atmospheric-pressure plasma jet (APPJ). METHODS AND RESULTS: Atmospheric pressure plasma jet was applied as mutagenic source for breeding high-yielding mutant strain. Helium was used as APPJ operating gas. The mutation effects of different treatment times of APPJ were studied. The mutant strain designated as B. subtilis B06 was successfully screened out, which showed higher antagonistic activity against F. graminearum in vitro. Its inhibition zone against the indicator fungus increased by 23% compared to the original one. HPLC and ESI (electrospray ionization) mass spectrometry analysis indicated that antifungal compounds produced by the mutant and original strain belonged to the lipopeptide, surfactin and iturin families. The mutant strain showed favourable properties of faster growth in the fermentation process and higher production of antibiotics. The lipopeptide production of the mutant was 2.3-fold as that of the original strain. CONCLUSIONS: A mutant strain with strong antagonistic activity and high yielding of antibiotics was obtained by APPJ in this study. The mutant could be used as a promising biocontrol agent in agriculture. SIGNIFICANCE AND IMPACT OF STUDY: This study provides a novel mutagenic source for breeding high-yielding microbial mutant, which would be very useful in the application of some valuable metabolites from micro-organism. PMID- 19558468 TI - Rapid enumeration of Escherichia coli in marine bathing waters: potential interference of nontarget bacteria. AB - AIMS: To compare the Escherichia coli quantification given by the 'Coliplage' assay, based on the direct measurement of the beta-D-glucuronidase (GLUase) activity and the reference Most Probable Number (MPN) method from seawater sites and investigate the possible interference of non-E. coli strains in the GLUase activity measurement. METHODS AND RESULTS: Comparison performed from 69 French coastal bathing sites (1401 samples) showed nonconcordance between both methods, only for 8% of samples. Non-E. coli 4-methylumbelliferyl-beta-D-glucuronide (MUG+) were isolated from nonconcordant samples. Phylogenetic analysis showed that Gammaproteobacteria were dominants and mainly represented by Vibrio species, which displayed GLUase activities on the same order of magnitude and sometimes much higher as E. coli reference strains. CONCLUSIONS: The'Coliplage' assay is a rapid method for the quantification of E. coli showed few discordances with the standard MPN method. Some Vibrio species could interfere on the direct GLUase activity measurement of E. coli. SIGNIFICANCE AND IMPACT OF THE STUDY: Data present the first qualitative investigation on disagreement between Coliplage and the MPN results. If the interference of Vibrio species is confirmed in situ, appropriate treatments should be developed to remove the interfering signal. PMID- 19558469 TI - Antifungal and marker effects of Talisia esculenta lectin on Microsporum canis in vitro. AB - AIMS: The purpose of this study was to demonstrate the usefulness of lectin obtained from Talisia esculenta (TEL) seeds as a tool to recognize and study Microsporum canis. For this purpose, we investigated the antifungal and marker action of this lectin and the relationship of these effects with the presence of carbohydrates on the structure of this fungus. METHODS AND RESULTS: The in vitro antifungal activity of TEL was analysed by broth microdilution assay. In addition, TEL was assessed against the arthroconidia present on hairs obtained from infected dogs and cats. The affinity of fluorescein isothiocyanate (FITC) labelled TEL for macroconidia and arthroconidia of M. canis was also tested. The effects of TEL on the growth of the M. canis strains began with 0.125 mg ml(-1), and 100% inhibition was obtained with a concentration of 2 mg ml(-1). The addition of carbohydrates, especially N-acetyl-glucosamine and d-mannose, inhibited these antifungal effects. TEL was able to inhibit the growth of arthroconidial chitin-rich forms of M. canis obtained from hairs of infected animals and strains cultured in Sabouraud agar. FITC-labelled TEL efficiently marked macroconidial and arthroconidial forms of M. canis, as shown by fluorescent microscopy. CONCLUSIONS: These results show that the inhibitory effects of TEL on M. canis growth may be related to the interaction of lectin with the carbohydrates present at the micro-organism's surface, mainly D-mannose and N-acetyl-glucosamine. SIGNIFICANCE AND IMPACT OF THE STUDY: Talisia esculenta can be used as an important tool in the biochemical study of M. canis or as a molecule to recognize this dermatophyte in infected tissue. PMID- 19558470 TI - Comparison of three plating media for the isolation of Salmonella from poultry environmental samples in Great Britain using ISO 6579:2002 (Annex D). AB - AIMS: To evaluate the performance of three Salmonella plating media (Rambach, Xylose Lysine Deoxycholate agar and modified Brilliant Green Agar plus Novobiocin) as part of the ISO 6579: 2002 (Annex D) on poultry environmental samples. METHODS AND RESULTS: The samples analysed were those for the European Union Salmonella baseline surveys of laying (N = 3087), broiler (N = 1550), turkey fattening (N = 1540) and turkey breeding (N = 580) flocks for Great Britain. Results were considered separately for Rambach (including and excluding pale orange colonies) and for growth on selective media [Modified semi-solid Rappaport Vassiliadis (MSRV)] after 24 and 48 h of incubation. Overall, Rambach was the most sensitive medium, provided that pale orange colonies were checked. In all cases, an increase in the sensitivity of detection was obtained by plating growth on MSRV after 48 h of incubation. In broilers and laying flocks, the specificity significantly improved when Rambach only was used. CONCLUSION: The use of Rambach results in considerable savings compared with the two-plate method prescribed by ISO 6579:2002 (Annex D) without compromising sensitivity. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella isolation protocols should be reviewed in terms of their efficiency and cost. PMID- 19558471 TI - Molecular typing of clinical Candida strains using random amplified polymorphic DNA and contour-clamped homogenous electric fields electrophoresis. AB - AIMS: This report describes an investigation into the genetic profiles of 38 Candida albicans and 19 Candida glabrata strains collected from a dental hospital of Monastir (Tunisia) and the Laboratory of Parasitology, Farhat Hached Hospital of Sousse (Tunisia), using two typing methods: random amplified polymorphic DNA (RAPD) and contour-clamped homogenous electric fields (CHEF). METHODS AND RESULTS: The two methods (RAPD and CHEF electrophoresis) were able to identify clonal-related isolates from different patients. RAPD method using two primers (CA1 and CA2) exhibited the highest discriminatory power by discriminating 22 genotypes for C. albicans with CA1 oligonucleotides and 19 genotypes with CA2 primer. For C. glabrata, 17 genotypes were obtained when both primers CA1 and CA2 were combined. The CHEF karyotyping of C. albicans has discriminated only 17 different karyotypes. CONCLUSION: The genotype of each isolate and genotypic difference among C. albicans and C. glabrata isolates were patient specific and not associated with the site of infection, geographic origin or date of isolation. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of relatedness between Candida species using molecular approaches with high discriminatory power is important in determining adequate measures for interruption of transmission of this yeast. PMID- 19558472 TI - Effect of cyclosporin A on functional recovery in the spinal cord following contusion injury. AB - Considerable evidence has shown that the immunosuppressant drug cyclosporin A (CsA) may have neuroprotective properties which can be exploited in the treatment of spinal cord injury. The aim of this study was to investigate the cellular environment within the spinal cord following injury and determine whether CsA has an effect on altering cellular interactions to promote a growth-permissive environment. CsA was administered to a group of rats 4 days after they endured a moderate contusion injury. Functional recovery was assessed using the Basso Beattie Bresnahan (BBB) locomotor rating scale at 3, 5 and 7 weeks post-injury. The rats were sacrificed 3 and 7 weeks post-injury and the spinal cords were sectioned, stained using histological and immunohistochemical methods and analysed. Using stereology, the lesion size and cellular environment in the CsA treated and control groups was examined. Little difference in lesion volume was observed between the two groups. An improvement in functional recovery was observed within CsA-treated animals at 3 weeks. Although we did not see significant reduction in tissue damage, there were some notable differences in the proportion of individual cells contributing to the lesion. CsA administration may be used as a technique to control the cell population of the lesion, making it more permissive to neuronal regeneration once the ideal environment for regeneration and the effects of CsA administration at different time points post injury have been identified. PMID- 19558473 TI - Sexual function and aging in racially and ethnically diverse women. AB - OBJECTIVES: To examine factors influencing sexual activity and functioning in racially and ethnically diverse middle-aged and older women. DESIGN: Cross sectional cohort study. SETTING: Integrated healthcare delivery system. PARTICIPANTS: One thousand nine hundred seventy-seven women aged 45 to 80. MEASUREMENTS: Self-administered questionnaires assessed sexual desire, activity, satisfaction, and problems. RESULTS: Of the 1,977 participants (876 white, 388 African American, 347 Latina, and 351 Asian women), 43% reported at least moderate sexual desire, and 60% had been sexually active in the previous 3 months. Half of sexually active participants (n=969) described their overall sexual satisfaction as moderate to high. Among sexually inactive women, the most common reason for inactivity was lack of interest in sex (39%), followed by lack of a partner (36%), physical problem of partner (23%), and lack of interest by partner (11%); only 9% were inactive because of personal physical problems. In multivariable analysis, African-American women were more likely than white women to report at least moderate desire (odds ratio (OR)=1.65, 95% confidence interval (CI)=1.25-2.17) but less likely to report weekly sexual activity (OR=0.68, 95% CI=0.48-0.96); sexually active Latina women were more likely than white women to report at least moderate sexual satisfaction (OR=1.75, 95% CI=1.20-2.55). CONCLUSION: A substantial proportion of community-dwelling women remain interested and engaged in sexual activity into older age. Lack of a partner capable of or interested in sex may contribute more to sexual inactivity than personal health problems in this population. Racial and ethnic differences in self-reported sexual desire, activity, and satisfaction may influence discussions about sexual difficulties in middle-aged and older women. PMID- 19558474 TI - Prevalence and characteristics of traumatic intracranial hemorrhage in elderly fallers presenting to the emergency department without focal findings. AB - OBJECTIVES: To determine the prevalence and associated characteristics of traumatic intracranial hemorrhage (ICH) in elderly fallers presenting to the emergency department (ED) without focal findings. DESIGN: Retrospective cohort study. SETTING: University-affiliated teaching hospital ED. PARTICIPANTS: Patients aged 65 and older presenting with a fall to the ED and undergoing a head computed tomography (CT) scan. MEASUREMENTS: Electronic medical records and CT scans of 404 consecutive patients were reviewed. Characteristics of patients with and without ICH were compared using unadjusted analyses. Patients taking warfarin, aspirin, or clopidogrel alone or in combination were compared with those not taking these medications. Multivariate logistic regression analyses were performed to determine variables independently associated with ICH. RESULTS: Forty-seven of 404 elderly fallers (11.6%) without focal findings had an ICH. Unadjusted analyses in these pilot data showed that warfarin was not significantly associated with ICH. Multivariate analyses indicated that elderly people living in at home were more likely to have ICH than those living in nursing homes or assisted living facilities (odds ratio (OR)=3.2, 95% confidence interval (CI)=1.30-8.13) and that those with head trauma were more likely to have ICH than those without (OR=3.9, 95% CI=1.25-7.80). Aspirin was found to be protective (OR=0.49, 95% CI=0.24-0.98). CONCLUSION: ICH is common in elderly fallers presenting to the ED without focal findings. Anticoagulation alone did not appear to increase the risk of ICH, and aspirin was found to be protective, but prospective studies are needed to better assess this relationship. PMID- 19558475 TI - The overlap syndrome of depression and delirium in older hospitalized patients. AB - OBJECTIVES: To measure the prevalence, predictors, and posthospitalization outcomes associated with the overlap syndrome of coexisting depression and incident delirium in older hospitalized patients. DESIGN: Secondary analysis of prospective cohort data from the control group of the Delirium Prevention Trial. SETTING: General medical service of an academic medical center. Follow-up interviews at 1 month and 1 year post-hospital discharge. PARTICIPANTS: Four hundred fifty-nine patients aged 70 and older who were not delirious at hospital admission. MEASUREMENTS: Depressive symptoms assessed at hospital admission using the 15-item Geriatric Depression Scale (cutoff score of 6 used to define depression), daily assessments of incident delirium from admission to discharge using the Confusion Assessment Method, activities of daily living at admission and 1 month postdischarge, and new nursing home placement and mortality determined at 1 year. RESULTS: Of 459 participants, 23 (5.0%) had the overlap syndrome, 39 (8.5%) delirium alone, 121 (26.3%) depression alone, and 276 (60.1%) neither condition. In adjusted analysis, patients with the overlap syndrome had higher odds of new nursing home placement or death at 1 year (adjusted odds ratio (AOR)=5.38, 95% confidence interval (CI)=1.57-18.38) and 1-month functional decline (AOR=3.30, 95% CI=1.14-9.56) than patients with neither condition. CONCLUSION: The overlap syndrome of depression and delirium is associated with significant risk of functional decline, institutionalization, and death. Efforts to identify, prevent, and treat this condition may reduce the risk of adverse outcomes in older hospitalized patients. PMID- 19558476 TI - Bruising as a marker of physical elder abuse. AB - OBJECTIVES: To describe bruising as a marker of physical elder abuse. DESIGN: Consenting older adults were examined to document location and size of bruises and assess whether they were inflicted during physical abuse. An expert panel confirmed physical abuse. Findings were compared with results of an earlier study of accidental bruising in older adults. SETTING: Residences of participants. PARTICIPANTS: Sixty-seven adults aged 65 and older reported to Adult Protective Services (APS) for suspected physical elder abuse. MEASUREMENTS: Age, sex, ethnicity, race, functional status, medical conditions, cognitive status, history of falls, bruise size and location, recall of cause, and responses to Revised Conflicts Tactics Scale and Elder Abuse Inventory. RESULTS: Seventy-two percent (n=48) of older adults who had been physically abused within 30 days before examination had bruises. The physically abused older adults had significantly larger bruises; more of them knew the cause of their bruises (43 (89.6%) vs 16 (23.5%) of the comparison group); and they were significantly more likely to have bruises on the face, lateral aspect of the right arm and the posterior torso (including back, chest, lumbar, and gluteal regions) than older adults from an earlier study who had not been abused (n=68). CONCLUSION: Bruises that occur as a result of physical elder mistreatment are often large (>5 cm) and on the face, lateral right arm, or posterior torso. Older adults with bruises should be asked about the cause of the bruises to help ascertain whether physical abuse occurred. PMID- 19558477 TI - Effect of inflammation in the periodontium in early old age on mortality at 21 year follow-up. AB - OBJECTIVES: To analyze whether inflammatory processes in the periodontium in early old age are related to subsequent mortality during 21 years of follow-up in a nondisabled 70-year-old population. SETTING: Community-based population in Copenhagen. DESIGN: The study was based on the Glostrup Aging Study of the 1914 population, with baseline in 1984 when the participants were 70 years old and follow-up 21 years later. PARTICIPANTS: Three hundred thirty-five dentate men and women participated in the clinical oral health examination. MEASUREMENTS: Severe periodontal inflammation was measured for all teeth present as the number of teeth with inflammation and periodontal pockets 6 mm deep or more. Mortality data were obtained from the Danish Death Register at 21-year follow-up. The Cox proportional hazards regression model was used. Covariates were measured at baseline and included number of teeth, caries, sex, education, income, hypertension, diabetes mellitus, osteoarthritis, arteriostenosis, myocardial infarction, comorbidity, fatigue, and ability to brush teeth. RESULTS: The analyses showed that severe periodontal inflammation in at least three teeth at age 70 was marginally related to mortality during 21-year follow-up (crude hazard ratio (HR)=1.17, 95% confidence interval (CI)=0.91-1.78). The estimate increased slightly when adjusted for sex, income, fatigue, and smoking (adjusted HR=1.37, 95% CI=0.97-1.92). The estimates were attenuated when adjusted for the specific diseases, especially arteriostenosis and osteoarthritis. CONCLUSION: Inflammation in the periodontium in early old age tends to be associated with mortality in older age. PMID- 19558478 TI - Risk factors for death in elderly emergency department patients with suspected infection. AB - OBJECTIVES: To identify independent risk factors for death in elderly emergency department (ED) patients admitted for infection and to derive and validate a mortality-prediction rule for such patients. DESIGN: Prospective cohort study. SETTING: Tertiary hospital ED with 55,000 annual visits. PARTICIPANTS: ED patients aged 65 and older admitted for infection between December 2003 and September 2004 in the derivation cohort and October 2005 and October 2006 in the validation cohort. MEASUREMENTS: PRIMARY OUTCOME: 28-day in-hospital mortality. Data were extracted from charts, and multivariate logistic regression were performed to identify independent mortality predictors. A prediction model was constructed and then validated in a second cohort. RESULTS: Nine hundred thirty five patients were included in the derivation cohort and 2,015 in the validation cohort. Mortality was 6% in the derivation cohort and 7% in the validation cohort. In the derivation cohort, logistic regression revealed five independent mortality predictors: respiratory compromise (respiratory rate >20 breaths per minute or hypoxemia) (odds ratio (OR)=4.0, 95% confidence interval (CI)=1.7-9.4), tachycardia (heart rate > or = 120 betas per minute; OR=3.2, 95% CI=1.6-6.3), cardiovascular failure (systolic blood pressure <90 mmHg despite fluid challenge or lactate > or = 4.0; OR=9.0, 95% CI=4.7-17), preexisting terminal illness (OR=5.7, 95% CI=2.2-15), and platelet count less than 150,000/mm3 (OR=2.7, 95% CI=1.3-5.6). Mortality increased with the number of factors: 0.51% for no factors, 3.1% for one factor, 14% for two factors, 47% for three or more risk factors. The c-statistic was 0.87 for the derivation model and 0.74 for the validation model. Almost 80% of patients in both cohorts were in low-risk groups (0 or 1 factor). CONCLUSION: A rule derived from five readily available variables predicts mortality in infected elderly ED patients and allows identification of a large low-risk subgroup. PMID- 19558479 TI - How will the U.S. healthcare system meet the challenge of the ethnogeriatric imperative? AB - Much of the geriatric imperative that is facing providers in the United States is an ethnogeriatric imperative, because one-third of older Americans are projected to be from one of the minority populations by mid-century, and that vastly underrepresents the actual diversity providers will see. Because of the vast heterogeneity of culture, language, health beliefs, risk for disease, and other factors, it is important for policy makers and health providers to be familiar with the diverse characteristics and needs of the various groups that will need geriatric care if they are to receive effective services. Challenges to high quality ethnogeriatric care include disparities in health status and health care, differences of acculturation level and other characteristics within the populations, language and limited English proficiency, health literacy, culturally defined health beliefs and syndromes, and specific beliefs and preferences about long-term and end-of-life care. Some models of successful ethnogeriatric care have been identified and have in common the involvement of members of the target population in the development and design of the services and the use of cultural liaisons from the ethnic community being served, such as community health workers, or promatores. Thirteen recommendations are suggested for policy and practice changes in multiethnic and ethnic-specific health programs to provide competent ethnogeriatric care in the U.S. healthcare system. PMID- 19558481 TI - Pain management interventions in the nursing home: a structured review of the literature. AB - Residents in nursing homes (NHs) experience pain that is underrecognized and undertreated. This pain contributes to a decline in quality of life. Although descriptive studies of pain assessment and management have been conducted, few have been published that critically evaluate interventions to improve pain management. Identification of the strengths and gaps in the current literature is required. A literature search was conducted of clinical trials that evaluated prospective interventions to improve pain management. Information on the intervention type, resident sample and setting, endpoints, and study design were extracted. Studies were classified based on a modification of Donabedian's model of healthcare quality. Four categories of interventions were identified: actor, decision support, treatment, and systems. The search strategy and selection criteria yielded 21 articles. Eleven studies used an actor intervention; of these, eight also employed a systems intervention, and one also used a treatment intervention. Two studies used a decision support intervention, seven used a treatment intervention, and one used a systems intervention. The overall quality of research was uneven in several areas: research design--nine studies were quasi experimental in nature, endpoints measures were not consistent--three did not perform statistical analysis, and characteristics of the resident samples varied dramatically. In conclusion, the number of high-quality studies of pain management in NHs remains limited. Process endpoints are used as surrogate measures for resident endpoints. Systematic approaches are needed to understand how each type of intervention improves the quality of pain management at the resident level. PMID- 19558480 TI - Serum insulin-like growth factor-1 binding proteins 1 and 2 and mortality in older adults: the Health, Aging, and Body Composition Study. AB - OBJECTIVE: To evaluate the relationship between serum insulin-like growth factor 1 (IGF-1), IGF-1 binding protein 1 (IGFBP-1), and IGF-1 binding protein 2 (IGFBP 2) and fasting insulin, fasting glucose, adiposity, and mortality in older adults. DESIGN: A prospective cohort study with mean follow-up of 6.2 years. SETTING: Participants were recruited and followed at two centers affiliated with academic medical institutions. PARTICIPANTS: Six hundred twenty-five men and women aged 70 and older and in good health at the time of enrollment. MEASUREMENTS: Serum IGF-1, IGFBP-1, and IGFBP-2; fasting serum insulin; fasting serum glucose; visceral fat; and total percent fat. RESULTS: Higher IGFBP-1 and higher IGFBP-2 were significantly associated with lower fasting insulin, lower fasting glucose, and lower adiposity, but higher IGFBP-1 and IGFBP-2 were associated with greater mortality. In multivariate adjusted models, the hazard ratio for all-cause mortality was 1.48 (95% confidence interval (CI)=1.14-1.92) per standard deviation (SD) increase in IGFBP-2 and 1.34 (95% CI=1.01-1.76) per SD increase in IGFBP-1. No association was found between IGF-1 and all-cause mortality. CONCLUSIONS: Higher IGFBP-1 and IGFBP-2 are associated with lower adiposity and decreased glucose tolerance but also with greater all-cause mortality. Higher levels of serum IGF-1 binding protein (IGFBP) may indicate greater IGF-1 activity and thus represent an association between higher IGF-1 activity and mortality in humans. PMID- 19558482 TI - Senior health clinics: are they financially viable? AB - Are hospital-based outpatient interdisciplinary clinics a financially viable alternative for caring for our burgeoning population of older adults in America? Although highly popular, with high patient satisfaction rates among older adults and their families, senior health clinics (SHCs) can be expensive to operate, with limited quantifiable health outcomes. This study analyzed three geriatric hospital-based interdisciplinary clinics in rural Arkansas by examining their patient profiles, revenues, and expenses. It closely examined the effects of the downstream revenue using the multiplier effect and acknowledged other factors that weigh heavily on the success of SHCs and the care of older adults. The findings highlight the similarities and differences in the three clinics' operating and financial structures in addition to the clinics' and providers' productivity. The analysis presents an evidence-based illustration that SHCs can break even or lose large amounts of money. PMID- 19558483 TI - Pilot testing of intervention protocols to prevent pneumonia in nursing home residents. AB - OBJECTIVES: To test intervention protocols for feasibility, staff adherence, and effectiveness in reducing pneumonia risk factors (impaired oral hygiene, swallowing difficulty) in nursing home residents. DESIGN: Prospective study. SETTING: Two nursing homes. PARTICIPANTS: Fifty-two nursing home residents. INTERVENTION: Thirty residents with impaired oral hygiene were randomly assigned to manual oral brushing plus 0.12% chlorhexidine oral rinse at different frequencies daily. Twenty-two residents with swallowing difficulty were randomly assigned to upright feeding positioning, teaching swallowing techniques, or manual oral brushing. All protocols were administered over 3 months. MEASUREMENTS: Feasibility was assessed monthly and defined as high if the protocol took less than 10 minutes to administer. Adherence was assessed weekly and defined as high if full staff adherence was demonstrated in more than 75% of assessments. Effectiveness for improved oral hygiene (reduction in oral plaque score) and swallowing (reduction in cough during swallowing) was compared at baseline and 3 months. RESULTS: Daily manual oral brushing plus 0.12% chlorhexidine rinse demonstrated high feasibility, high staff adherence, and effectiveness in improving oral hygiene (P<.001 vs baseline); this combination administered twice per day showed the highest plaque score reduction. Daily manual oral brushing and upright feeding positioning demonstrated high feasibility, high staff adherence, and effectiveness in improving swallowing. CONCLUSION: Manual oral brushing, 0.12% chlorhexidine oral rinse, and upright feeding positioning demonstrated high feasibility, high staff adherence, and effectiveness in pneumonia risk factor reduction. A protocol combining these components warrants testing for its ability to reduce pneumonia in nursing home residents. PMID- 19558485 TI - Care of the hospitalized aging medical patient: delirium module. PMID- 19558484 TI - A cluster-randomized trial of an educational intervention to reduce the use of physical restraints with psychogeriatric nursing home residents. AB - OBJECTIVES: To investigate the effects of an educational intervention on the use of physical restraints with psychogeriatric nursing home residents. DESIGN: Cluster-randomized trial. SETTING: Fifteen psychogeriatric nursing home wards in the Netherlands. PARTICIPANTS: In total, 432 psychogeriatric nursing home residents from 15 psychogeriatric nursing home wards in seven nursing homes were selected for participation; 404 consented, and 371 of these were available at baseline. Two hundred forty-one from 14 wards had complete data and were included in the data analyses. INTERVENTION: The nursing home wards were assigned at random to educational intervention or control status. The educational intervention consisted of an educational program for nursing staff combined with consultation with a nurse specialist (registered nurse (RN) level). MEASUREMENTS: Data were collected at baseline and 1, 4, and 8 months postintervention. At each measurement, the use of physical restraints was measured using observations of blinded, trained observers on four separate occasions over a 24-hour period. Other resident characteristics, such as cognitive status, were determined using the Minimum Data Set. RESULTS: Logistic and linear regression analyses showed no treatment effect on restraint status, restraint intensity, or multiple restraint use in any of the three postintervention measurements. Furthermore, only small changes occurred in the types of restraints used with residents in the experimental group. CONCLUSION: An educational program for nursing staff combined with consultation with a nurse specialist (RN level) had no effect on the use of physical restraints with psychogeriatric nursing home residents. In addition to restraint education and consultation, new measures to reduce the use of physical restraints with psychogeriatric nursing home residents should be developed. PMID- 19558486 TI - Intramuscular olanzapine vs. intramuscular short-acting antipsychotics: safety, tolerability and the switch to oral antipsychotic medication in patients with schizophrenia or acute mania. AB - BACKGROUND: This study compared the safety, tolerability and switch to oral medication in patients with bipolar disorder or schizophrenia who received intramuscular (IM) olanzapine or other IM antipsychotics for the treatment of acute agitation. METHODS: Patients (N = 2011) from 15 countries participated in this prospective, observational, non-interventional study. Inpatients requiring treatment with at least one IM injection of a short-acting antipsychotic were assessed at baseline and within 7 days after the first IM injection. Treatment groups comprised: (i) patients prescribed IM olanzapine at baseline; and (ii) patients prescribed any other IM antipsychotic medication at baseline. Outcome measures included: treatment-emergent adverse events, concomitant psychotropic medication and the time taken to switch to oral medication. RESULTS: Fewer patients in the IM olanzapine group experienced an adverse event than patients in the other IM antipsychotic group (34.4% vs. 46.2%, p < 0.001). The most frequently reported adverse events in both groups were: sedation, Parkinsonism, disturbance in attention, akathisia, dystonia and orthostatic hypotension. Fewer patients in the IM olanzapine group used anticholinergics (13.9% vs. 42.5%, p < 0.001) or anxiolytics/hypnotics (47.6% vs. 51.6%, p = 0.023). Patients in the IM olanzapine group switched to oral medication earlier than patients in the other IM antipsychotic group (median time = 46.5 vs. 48.0 h, p = 0.009). CONCLUSIONS: These findings suggest that IM olanzapine may have a favourable impact on individual patients. However, the high rate of oral concomitant medication used throughout the study limits these findings from being associated with IM olanzapine alone. PMID- 19558487 TI - Can simple tests performed in the primary care setting provide accurate and efficient diagnosis of benign prostatic hyperplasia? Rationale and design of the Diagnosis Improvement in Primary Care Trial. AB - Effective treatment of benign prostatic hyperplasia (BPH) improves lower urinary tract symptoms (LUTS) and patient quality of life, and reduces the risk of complications arising from disease progression. However, treatment can only be initiated when men with BPH are identified by accurate diagnostic tests. Current evidence suggests that diagnostic procedures employed by primary care physicians vary widely across Europe. The expected increases in BPH prevalence accompanying the gradual aging of the population, coupled with greater use of medical therapy, mean that general practitioners (GPs) are likely to have an increasingly important role in managing the condition. The GP/primary care clinic is therefore an attractive target location for strategies designed to improve the accuracy of BPH diagnosis. The Diagnosis Improvement in Primary Care Trial (D-IMPACT) is a prospective, multicentre, epidemiological study that aims to identify the optimal subset of simple tests applied by GPs in the primary care setting to diagnose BPH in men who spontaneously report obstructive (voiding) and/or irritative (storage) LUTS. These tests comprise medical history, symptom assessment with the International Prostate Symptom Score questionnaire, urinalysis, measurement of serum levels of prostate-specific antigen and subjective GP diagnosis after completing all tests including digital rectal examination. GP diagnoses and all other tests will be compared with gold-standard diagnoses provided by specialist urologists following completion of additional diagnostic tests. D-IMPACT will establish the diagnostic performance using a non-subjective and reproducible algorithm. An adjusted and multivariate analysis of the results of D-IMPACT will allow identification of the most efficient combination of tests that facilitate accurate BPH diagnosis in the primary care setting. In addition, D-IMPACT will estimate the prevalence of BPH in patients who present spontaneously to GPs with LUTS. PMID- 19558488 TI - Bypassing agent regimens and costs for prophylaxis in patients with inhibitors. PMID- 19558489 TI - Uneventful coronary artery bypass surgery without prophylactic replacement therapy in a patient with a novel heterozygous FVII gene deletion. PMID- 19558490 TI - Engineering of monomeric FK506-binding protein 22 with peptidyl prolyl cis-trans isomerase. Importance of a V-shaped dimeric structure for binding to protein substrate. AB - FK506-binding protein 22 (FKBP22) from the psychrotrophic bacterium Shewanella sp. SIB1 is a homodimeric protein with peptidyl prolyl cis-trans isomerase (PPIase) (EC 5.2.1.8) activity. Each monomer consists of 205 amino acid residues. According to a tertiary model, SIB1 FKBP22 assumes a V-shaped structure, in which two monomers interact with each other at their N-termini. Each monomer consists of an N-terminal domain with a dimerization core and a C-terminal catalytic domain, which are separated by a 40-residue-long a-helix. To clarify the role of this V-shaped structure, we constructed a mutant protein, in which the N-domain is tandemly repeated through a flexible linker. This protein, termed NNC-FKBP22, is designed such that two repetitive N-domains are folded into a structure similar to that of the Shewanella sp. SIB1 FKBP22 wild-type protein (WT). NNC FKBP22 was overproduced in Escherichia coli in a His-tagged form, purified and biochemically characterized. Gel-filtration chromatography and ultracentrifugation analyses indicate that NNC-FKBP22 exists as a monomer. Analysis of thermal denaturation using differential scanning calorimetry indicates that NNC-FKBP22 unfolds with two transitions, as does the WT protein. NNC-FKBP22 exhibited PPIase activity for both peptide and protein substrates. However, in contrast to its activity for peptide substrate, which was comparable to that of the WT protein, its activity for protein substrate was reduced by five to six-fold, compared to that of the WT. Surface plasmon resonance analyses indicate that NNC-FKBP22 binds to a reduced form of a-lactalbumin with a six-fold weaker affinity than that of WT. These results suggest that a V-shaped structure of SIB1 FKBP22 is important for efficient binding to a protein substrate. PMID- 19558491 TI - Design and humanization of a murine scFv that blocks human platelet glycoprotein VI in vitro. AB - Platelet adhesion and aggregation at the site of vascular injury is essential for hemostasis, but can also lead to arterial occlusion in thrombotic disorders. Glycoprotein (GP) VI is the major platelet membrane receptor that interacts directly with collagen, the most thrombogenic compound in the blood vessels. GPVI could therefore be a major therapeutic target. Fab fragments of the anti-GPVI murine monoclonal IgG 9O12 have previously been shown to completely block collagen-induced platelet aggregation, to inhibit the procoagulant activity of collagen-stimulated platelets, and to prevent thrombus formation under arterial flow conditions without significantly prolonging the bleeding time. Here, we engineered recombinant scFvs that preserve the functional properties of 9O12, and could constitute building blocks for designing new compounds with potentially therapeutic antithrombotic properties. First, the 9O12 variable domains were cloned, sequenced, and expressed as a recombinant murine scFv, which was fully characterized. This scFv preserved all the characteristics that make 9O12 Fab potentially useful for therapeutic applications, including its high affinity for GPVI, ability to inhibit platelet adhesion, and aggregation with collagen under arterial flow conditions. A humanized version of this scFv was also designed after complementarity-determining region grafting and structural refinements using homology-based modeling. The final product was produced in recombinant bacteria. It retained GPVI-binding specificity and high affinity, which are the main parameters usually impaired by humanization procedures. This is a simple, efficient and straightforward method that could also be used for humanizing other antibodies. PMID- 19558492 TI - Structure of RNase Sa2 complexes with mononucleotides--new aspects of catalytic reaction and substrate recognition. AB - Although the mechanism of RNA cleavage by RNases has been studied for many years, there remain aspects that have not yet been fully clarified. We have solved the crystal structures of RNase Sa2 in the apo form and in complexes with mononucleotides. These structures provide more details about the mechanism of RNA cleavage by RNase Sa2. In addition to Glu56 and His86, which are the principal catalytic residues, an important role in the first reaction step of RNA cleavage also seems to be played by Arg67 and Arg71, which are located in the phosphate binding site and form hydrogen bonds with the oxygens of the phosphate group of the mononucleotides. Their positive charge very likely causes polarization of the bonds between the oxygens and the phosphorus atom, leading to electron deficiency on the phosphorus atom and facilitating nucleophilic attack by O2' of the ribose on the phosphorus atom, leading to cyclophosphate formation. The negatively charged Glu56 is in position to attract the proton from O2' of the ribose. Extended molecular docking of mononucleotides, dinucleotides and trinucleotides into the active site of the enzyme allowed us to better understand the guanosine specificity of RNase Sa2 and to predict possible binding subsites for the downstream base and ribose of the second and third nucleotides. PMID- 19558493 TI - Investigation and prediction of the severity of p53 mutants using parameters from structural calculations. AB - A method has been developed to predict the effects of mutations in the p53 cancer suppressor gene. The new method uses novel parameters combined with previously established parameters. The most important parameter is the stability measure of the mutated structure calculated using molecular modelling. For each mutant, a severity score is reported, which can be used for classification into deleterious and nondeleterious. Both structural features and sequence properties are taken into account. The method has a prediction accuracy of 77% on all mutants and 88% on breast cancer mutations affecting WAF1 promoter binding. When compared with earlier methods, using the same dataset, our method clearly performs better. As a result of the severity score calculated for every mutant, valuable knowledge can be gained regarding p53, a protein that is believed to be involved in over 50% of all human cancers. PMID- 19558494 TI - Inhibition of 12-LOX and COX-2 reduces the proliferation of human epidermoid carcinoma cells (A431) by modulating the ERK and PI3K-Akt signalling pathways. AB - Eicosanoids, the oxygenated metabolites of arachidonic acid (AA), mediate a variety of human diseases, such as cancer, inflammation and arthritis. To evaluate the role of eicosanoids in epidermoid carcinoma, the expression of AA metabolizing enzymes, such as lipoxygenases (LOXs) and cyclooxygenases (COXs), was analysed in a human epidermoid carcinoma cell line (A431). These studies revealed overexpression of 12-R-LOX and COX-2 in A431 cells. Baicalein (a 12-LOX inhibitor) and celecoxib (a COX-2 inhibitor) significantly reduced thymidine incorporation, whereas 12-(R)-HETE and 12-(S)-HETE (12-LOX metabolites) and PGE(2) (COX-2 metabolite) significantly enhanced thymidine incorporation, suggesting a role for these enzymes in the regulation of A431 cell proliferation. Further studies on the mechanism of cell death by baicalein and celecoxib revealed that the induction of apoptosis in A431 cells was associated with reduction in the Bcl-2/Bax ratio, release of cytochrome c, activation of caspase 3 and PARP cleavage. The apoptosis induced by baicalein and celecoxib was mediated by down regulation of ERK and PI3K-Akt pathways. Further, 12-(R)-HETE, 12-(S)-HETE and PGE(2) upregulated the p-ERK and p-Akt levels, suggesting the involvement of ERK and Akt pathways in the 12-LOX- and COX-2-mediated regulation of growth in A431 cells. Our findings suggest that 12-R-LOX and COX-2 play a critical role in the regulation of growth in epidermoid carcinoma and that their inhibitors may be of potential therapeutic importance. PMID- 19558495 TI - An improved method of human keratinocyte culture from skin explants: cell expansion is linked to markers of activated progenitor cells. AB - Human keratinocyte primary cultures are commonly established by tissue dissociation and often rely on feeder cell supports and culture medium that is not defined. Further, contamination by unwanted fibroblasts can be problematic. Here, we developed a skin explant method for growing primary keratinocytes that was rapid, simple, and reliably generated keratinocyte cultures free of fibroblast contamination. The process capitalized on the observation that fibroblasts migrate out of adult skin explants later than epidermal cells, allowing the early harvesting of keratinocytes by trypsinization. When grown subsequently in defined medium in the absence of feeder cells, the explant derived cells grew rapidly and could be cultured for multiple passages. Immunofluorescence microscopy revealed that a high percentage of cells harvested from the explant outgrowths expressed K15, while very few expressed the differentiation marker K10. Cells that were stained while migrating out from explants strongly expressed markers associated with progenitor cells, including p63, K15 and CD133, and displayed intense K6 expression, indicative of activated keratinocytes in wound-healing epidermis. By replenishing the explants with fresh medium after harvesting, further epidermal outgrowths could be obtained, offering the possibility of greatly increased keratinocyte yields for clinical applications. PMID- 19558496 TI - Sulforaphane but not ascorbigen, indole-3-carbinole and ascorbic acid activates the transcription factor Nrf2 and induces phase-2 and antioxidant enzymes in human keratinocytes in culture. AB - Nrf2 is a basic leucine zipper transcriptional activator essential for the coordinated transcriptional induction of phase-2 and antioxidant enzymes. Brassica vegetables contain phytochemicals including glucoraphanin, the precursor of sulforaphane (SFN) and glucobrassicin, the precursor of indole-3-carbinole (I3C) and ascorbigen (ABG). The degradation products SFN, I3C and ABG may be capable of inducing cytoprotective genes in skin. In this study, we tested the potency of SFN, ABG and I3C in affecting Nrf2-dependent gene expression in human keratinocytes in culture. SFN but not ABG and its precursors I3C and ascorbic acid induced Nrf2 dependent gene expression at a relatively low concentration (5 micromol/l). Induction of Nrf2 due to SFN was accompanied by an increase in mRNA and protein levels of NADPH quinone oxidoreductase 1, heme oxygenase 1 and gamma glutamylcysteine-synthetase. Furthermore, SFN elevated cellular glutathione levels and antagonized tumor necrosis factor-alpha-induced NFkappaB transactivation. Therefore, SFN treatment may present a strategy for enhancing the cellular defense mechanisms in skin. PMID- 19558497 TI - Osteopontin and the skin: multiple emerging roles in cutaneous biology and pathology. AB - Osteopontin (OPN) is a glycoprotein expressed by various tissues and cells. The existence of variant forms of OPN as a secreted (sOPN) and intracellular (iOPN) protein and its modification through post-translational modification and proteolytic cleavage explain its broad range of functions. There is increasing knowledge which receptors OPN isoforms can bind to and which signaling pathways are activated to mediate different OPN functions. sOPN interacts with integrins and CD44, mediates cell adhesion, migration and tumor invasion, and has T helper 1 (Th1) cytokine functions and anti-apoptotic effects. iOPN has been described to regulate macrophage migration and interferon-alpha secretion in plasmacytoid dendritic cells. Both sOPN and iOPN, through complex functions for different dendritic cell subsets, participate in the regulation of Th cell lineages, among them Th17 cells. For skin disease, OPN from immune cells and tumor cells is of pathophysiological relevance. OPN is secreted in autoimmune diseases such as lupus erythematosus, and influences inflammation of immediate and delayed type allergies and granuloma formation. We describe that OPN is overexpressed in psoriasis and propose a model to study OPN function in psoriatic inflammation. Through cytokine functions, OPN supports immune responses against Mycobacteria and viruses such as herpes simplex virus. OPN is also implicated in skin tumor progression. Overexpression of OPN influences invasion and metastasis of melanoma and squamous cell carcinoma cells, and OPN expression in melanoma is a possible prognostic marker. As OPN protein preparations and anti-OPN antibodies may be available in the near future, in-depth knowledge of OPN functions may open new therapeutic approaches for skin diseases. PMID- 19558499 TI - Skin and heart: une liaison dangereuse. AB - Both skin and heart are subject to shear mechanical stress and need to be stress resistant in a flexible way. The intercellular connecting structures in skin and heart, the desmosomes, that have to resist these forces show remarkable resemblance in epidermis and myocardium. Mutations in desmosomal proteins lead to inherited desmosomal cardiocutaneous syndromes (DCCS): une liaison dangereuse. This article will critically review the cutaneous and cardiac features as well as the molecular background of DCCS, such as Naxos disease and Carvajal syndrome caused by deficiencies of plakoglobin and desmoplakin respectively. In addition, potential other desmosomal gene candidates for an involvement in cardiocutaneous syndromes are considered. The skin features in these syndromes may be the hallmark for the presence of progressive and ultimately lethal cardiac disease. Knowledge of these skin features and early recognition of such a syndrome may provide opportunities to halt or slow down cardiac disease progression, treat arrhythmias and even prevent sudden death. PMID- 19558498 TI - Proliferative defects in dyskeratosis congenita skin keratinocytes are corrected by expression of the telomerase reverse transcriptase, TERT, or by activation of endogenous telomerase through expression of papillomavirus E6/E7 or the telomerase RNA component, TERC. AB - Dyskeratosis congenita (DC) is characterized by the triad of reticulate skin pigmentation, nail dystrophy and leukoplakia. Epidermal atrophy, hair growth defects, bone marrow failure and increased risk of cancer are also common in DC patients. DC is caused by mutations in genes encoding for telomerase complex factors. Although there is an association of epidermal abnormalities with DC, epidermal cells from DC donors have not been previously characterized. We have isolated skin keratinocytes from affected members of a family with an autosomal dominant form of DC that is caused by a mutation in the RNA component of telomerase, TERC. Here, we demonstrate that, similar to DC fibroblasts from these donors, DC keratinocytes have short telomeres and a short lifespan. DC keratinocytes also exhibited impaired colony forming efficiency (CFE) and migration capacity. Exogenous expression of the reverse transcriptase (RT) component of telomerase, TERT, activated telomerase levels to half that of TERT expressing normal cells and maintained telomeres at a short length with concomitant extension of lifespan. Unlike fibroblasts, transduction of human papillomavirus type 16 E6/E7 genes into DC keratinocytes activated telomerase to half that of E6/E7 expressing normal cells, and robust proliferation was observed. While expression of TERC has no measurable effect on telomerase in fibroblasts, expression of TERC in keratinocytes upregulated telomerase activity and, rarely, allowed rescue of proliferative defects. Our results point to important differences between DC fibroblasts and keratinocytes and show, for the first time, that expression of TERC can increase the lifespan of primary human epithelial cells. PMID- 19558500 TI - Induction of eosinophil- and Th2-attracting epidermal chemokines and cutaneous late-phase reaction in tape-stripped skin. AB - Skin barrier damage induces various harmful or even protective reactions in the skin, as represented by enhancement of keratinocyte cytokine production. To investigate whether acute removal of stratum corneum modulates the production of chemokines by epidermal cells, we treated ears of BALB/c and C57BL/6 mice by tape stripping, or acetone-rubbing as a control of acute barrier disruption procedure. There was no difference between the tape-stripped and acetone-rubbed skin sites in the increased and recovered levels of transepidermal water loss. The mRNA expression levels of all the chemokines tested, including Th1 chemokines (CXCL10, CXCL9 and CXCL11), Th2 chemokines (CCL17 and CCL22) and eosinophil chemoattractant (CCL5), were higher in the epidermal cells from BALB/c than in those of C57BL/6 mice. In particular, CCL17, CCL22 and CCL5 were remarkably elevated in BALB/c mice and augmented by tape-stripping more markedly than acetone-rubbing, whereas Th1 chemokines were enhanced by acetone-rubbing more remarkably. Tape-stripping induced dermal infiltration of eosinophils in BALB/c but not C57BL/6 mice. In a contact hypersensitivity model, where BALB/c mice were sensitized on the abdomen and challenged on the ears with fluorescein isothiocyanate, mice exhibited higher ear swelling responses at the late-phase as well as delayed-type reactions, when challenged via the tape-stripped skin. The challenge via tape-stripped skin augmented the expression of IL-4 and CCR4 in the skin homogenated samples, indicating infiltration of Th2 cells. These findings suggest that acute barrier removal induces the expression of Th2 and eosinophil chemokines by epidermal cells and easily evokes the late phase reaction upon challenge with antigen. PMID- 19558502 TI - Effect of synthetic vernix biofilms on barrier recovery of damaged mouse skin. AB - The aim of this work was to investigate whether topical application of synthetic biofilms supports and accelerates the recovery of the murine skin barrier, disrupted by sequential tape stripping. Therefore, various biofilms were applied topically on disrupted mouse skin to determine which formulation could improve barrier function, as was observed previously for the natural biofilm vernix caseosa (VC). The biofilms [i.e. particles (synthetic corneocytes) embedded in a synthetic lipid matrix] mimic closely the physicochemical properties and structure of VC. Various formulations were prepared using different particle:lipid ratios, particles with different initial water content and uncoated or lipid-coated particles. It was observed that application of all tested formulations improved the skin barrier recovery rate and reduced crust formation and epidermal hyperproliferation. However, only one of the biofilms [i.e. B1; composed of uncoated particles with 50% (w/w) initial water content and particle:lipid ratio of 2:1] mimicked the effects of native VC most closely. This indicates the importance of the presence of individual components, i.e. barrier lipids and water, as well as the ratio of these components. Consequently, these observations suggest the potential use of this biofilm treatment clinically. PMID- 19558501 TI - Neuroendocrine activity of the melanocyte. AB - More than 15 years ago, we have proposed that melanocytes are sensory and regulatory cells with computing capability, which transform external and/or internal signals/energy into organized regulatory network(s) for the maintenance of the cutaneous homeostasis. This concept is substantiated by accumulating evidence that melanocytes produce classical stress neurotransmitters, neuropeptides and hormones, express corresponding receptors and these processes are modified and/or regulated by ultraviolet radiation, biological factors or stress. Examples of the above are catecholamines, serotonin, N-acetyl-serotonin, melatonin, proopiomelanocortin-derived adrenocorticotropic hormone, beta endorphin or melanocyte-stimulating hormone peptides, corticotropin releasing factor, related urocortins and corticosteroids including cortisol and corticosterone as well as their precursors. Furthermore, their production is not random, but hierarchical and follows the structures of classical neuroendocrine organizations such as hypothalamic-pituitary-adrenal axis, serotoninergic, melatoninergic and catecholaminergic systems. An example of an intrinsic but overlooked neuroendocrine activity is production and secretion of melanogenesis intermediates including l-DOPA or its derivatives that could enter circulation and act on distant sites. Such capabilities have defined melanocytes as neuroendocrine cells that not only coordinate cutaneous but also can affect a global homeostasis. PMID- 19558503 TI - Identification of new sensitive biomarkers for the in vivo response to interferon beta treatment in multiple sclerosis using DNA-array evaluation. AB - OBJECTIVE: Neutralizing antibodies (NAbs) occur in a proportion of multiple sclerosis (MS) patients treated with interferon (IFN)-beta. NAbs impair the effect of treatment. The biological effect of IFN-beta can be measured as the induction of the myxovirus resistance protein A (MxA) molecule. However, other markers could be more sensitive for evaluating the response to IFN-beta. We used DNA array analysis to identify genes that are strongly induced in blood cells by IFN-beta, and measured their expression in MS patients with different NAb levels. METHODS: Gene expression was studied on DNA arrays in untreated patients, in NAb negative patients, and in MS patients with varying NAb levels 9-12 h and 36-48 h after IFN-beta administration. The expression of selected genes was measured by real-time PCR. NAb levels were assessed by a cytopathic effect assay. RESULTS: Several hundred genes were induced 9-12 h after an injection of IFN-beta. The molecules CXCL10, CCL2 and IFI27 were among the most strongly induced. Gene induction was generally much less pronounced after 36-48 h, but IFI27 remained strongly induced. The strong induction of these molecules and MxA was confirmed by real-time PCR. Induction of MxA, CCL2, CXCL10 and IFI27 was reduced in patients with low NAb levels and lost in patients with intermediate/high NAb levels. CONCLUSION: We identify IFI27, CCL2 and CXCL10 as sensitive biomarkers for the response to IFN-beta. The expression of these markers adequately reflects bioactivity of IFN-ss as documented by the decreased induction in low NAb positive patients and the lost induction in patients with moderate/high NAb levels. PMID- 19558505 TI - Increased expression of TF on monocytes, but decreased numbers of TF bearing microparticles in blood from patients with acute myocardial infarction. PMID- 19558504 TI - Primary cutaneous diffuse large B-cell lymphoma, leg type, with features simulating POEMS syndrome. AB - A 91-year-old woman presented with a rapidly proliferative cutaneous lesion on the left lower limb, which was identified as a primary cutaneous diffuse large B cell lymphoma (PCLBCL), leg type, on biopsy. The patient also showed complications of hepatomegaly, endocrinopathy, edema, skin change, and polyneuropathy without monoclonal plasma cell proliferative disorder, and was therefore diagnosed with POEMS-like syndrome owing to the lack of monoclonal plasma cell proliferative disorder. Levels of serum vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) were high with the lymphoma cells immunostained positively for VEGF and IL-6. To the best of our knowledge, this is the first case report of PCLBCL, leg type, with POEMS-like syndrome. The findings in this case suggest that the symptoms of POEMS-like syndrome might be caused by the cytokines produced by the lymphoma cells. Furthermore, a wider range of diagnostic criteria associated with the result of abnormal secretion of cytokine may have to be considered for the diagnosis and evaluation of patients with possible POEMS syndrome, as against the present criteria specifying monoclonal plasma cell proliferative disorder as the essential criterion. PMID- 19558506 TI - Two independent gene signatures in pediatric t(4;11) acute lymphoblastic leukemia patients. AB - OBJECTIVE: Gene expression profiles become increasingly more important for diagnostic procedures, allowing clinical predictions including treatment response and outcome. However, the establishment of specific and robust gene signatures from microarray data sets requires the analysis of large numbers of patients and the application of complex biostatistical algorithms. Especially in case of rare diseases and due to these constrains, diagnostic centers with limited access to patients or bioinformatic resources are excluded from implementing these new technologies. METHOD: In our study we sought to overcome these limitations and for proof of principle, we analyzed the rare t(4;11) leukemia disease entity. First, gene expression data of each t(4;11) leukemia patient were normalized by pairwise subtraction against normal bone marrow (n = 3) to identify significantly deregulated gene sets for each patient. RESULT: A 'core signature' of 186 commonly deregulated genes present in each investigated t(4;11) leukemia patient was defined. Linking the obtained gene sets to four biological discriminators (HOXA gene expression, age at diagnosis, fusion gene transcripts and chromosomal breakpoints) divided patients into two distinct subgroups: the first one comprised infant patients with low HOXA genes expression and the MLL breakpoints within introns 11/12. The second one comprised non-infant patients with high HOXA expression and MLL breakpoints within introns 9/10. CONCLUSION: A yet homogeneous leukemia entity was further subdivided, based on distinct genetic properties. This approach provided a simplified way to obtain robust and disease-specific gene signatures even in smaller cohorts. PMID- 19558507 TI - Prognostic relevance of cytogenetic abnormalities in primary myelofibrosis: comparison of recent reports from Japan, the Mayo Clinic and MD Anderson Cancer Center. PMID- 19558508 TI - Health-related quality of life assessment in randomised controlled trials in multiple myeloma: a critical review of methodology and impact on treatment recommendations. AB - OBJECTIVES: Patients with multiple myeloma (MM) often have pronounced symptoms and substantially reduced quality of life. The aims of treatment are to control disease, maximise quality of life and prolong survival. Hence, health-related quality of life (HRQOL) should be an important end-point in randomised controlled trials (RCTs) in addition to traditional endpoints. We wanted to evaluate whether trials reporting HRQOL outcomes have influenced clinical decision making and whether HRQOL was assessed robustly according to predefined criteria. METHODS: A systematic review identified RCTs in MM with HRQOL assessment as a study end point. The methodological quality of these studies was assessed according to a checklist developed for evaluating HRQOL outcomes in clinical trials. The impact of the HRQOL results on clinical decision making was assessed, using published clinical guidelines as a reference. RESULTS: Fifteen publications presenting RCTs with HRQOL as a study end-point were identified. In 13 trials, the author stated that HRQOL results should influence clinical decision making. We found, however, that the HRQOL data only had a limited impact on published treatment guidelines for bisphosphonates, high-dose treatment, interferon, erythropoiesis-stimulating agents and novel agents. CONCLUSION: The present review indicates that the there are still few RCTs in MM including HRQOL as a study end-point. Systematic incorporation of HRQOL measures into clinical trials allows for a comparison of treatment arms that includes the patients' perspective. Before the full impact on clinical decisions can be realised, the quality and methodology of collecting HRQOL data must be further improved and the results rendered more comprehensible to clinicians. PMID- 19558509 TI - The inhibitory effect of resveratrol on leptin secretion from rat adipocytes. AB - BACKGROUND: Resveratrol was found to alleviate consequences of some metabolic disturbances which may be due to inappropriate dietary habits. It decreases mortality, increases insulin sensitivity and improves motor functions; these effects are accompanied by reduced plasma leptin and insulin. Leptin plays a significant role in the regulation of food intake and energy expenditure - elevated level in blood is one of the reasons of leptin-resistance and obesity. In this study, the direct effect of resveratrol on leptin secretion from isolated adipocytes was investigated. MATERIAL AND METHODS: Isolated rat adipocytes were incubated with resveratrol (62.5, 125 or 250 microM) and its effects on leptin secretion were studied. Cells were incubated with resveratrol in the presence of glucose (5 and 20 mM) and insulin (10 nM); glucose and nicotinic acid (1 mM); glucose and insulin in the presence of an inhibitor of protein kinase A (H-89, 50 microM) or alanine (10 mM) and insulin. The glucose uptake, glycerol release to the incubation medium, lactate and ATP produced by the cells were also measured. RESULTS: Resveratrol inhibited leptin secretion in all experimental designs in a dose-dependent manner. The effect was not accompanied by changes in glycerol release and glucose uptake. Adipocyte exposure to resveratrol enhanced the lactate formation. It was found that resveratrol dramatically reduced ATP in adipocytes. CONCLUSION: The obtained results revealed the direct ability of resveratrol to reduce leptin secretion from isolated rat adipocytes. Resveratrol is therefore a compound affecting the endocrine function of adipocytes. PMID- 19558510 TI - Quantitative PCR reveals transient and persistent algal viruses in Lake Ontario, Canada. AB - To determine if different algal viruses (Phycodnaviridae) share common patterns of seasonal abundance, quantitative PCR methods were developed and applied to monitor the abundances of three different viruses in Lake Ontario, Canada over 13 months. Throughout the year, the abundances of two different phycodnavirus polB gene fragments (LO1b-49 and LO1a-68) varied by more than two orders of magnitude, peaked during the autumn months, and were lowest during the summer. The seasonal abundance patterns of these two virus genes were similar and both were detected in almost every sample, but LO1b-49 was consistently an order of magnitude more abundant than LO1a-68. LO1b-49 reached a maximum abundance of 5413 +/- 312 genes ml(-1), whereas LO1a-68's abundance peaked at only 881 +/- 113 genes ml(-1). Another phycodnavirus polB fragment that was monitored (LO1b-16) was detected in only a few samples, but reached a higher maximum concentration (6771 +/- 879 genes ml(-1)) than either LO1b-49 or LO1a-68. The results of this year-long investigation of virus gene abundances suggests that Lake Ontario's phycodnavirus community is composed of persistent viruses detectable throughout the year and transient viruses present in only a few sporadic samples. The results also suggest that some persistent algal viruses are able to survive at relatively low abundances through several seasons. PMID- 19558511 TI - Viral control of bacterial biodiversity--evidence from a nutrient-enriched marine mesocosm experiment. AB - We demonstrate here results showing that bottom-up and top-down control mechanisms can operate simultaneously and in concert in marine microbial food webs, controlling prokaryote diversity by a combination of viral lysis and substrate limitation. Models in microbial ecology predict that a shift in the type of bacterial growth rate limitation is expected to have a major effect on species composition within the community of bacterial hosts, with a subsequent shift in the composition of the viral community. Only moderate effects would, however, be expected in the absolute number of coexisting virus-host pairs. We investigated these relationships in nutrient-manipulated systems, under simulated in situ conditions. There was a strong correlation in the clustering of the viral and bacterial community data supporting the existence of an important link between the bacterial and viral communities. As predicted, the total number of viral populations was the same in all treatments, while the composition of the viral community varied. Our results support the theoretical prediction that there is one control mechanism for the number of niches for coexisting virus-host pairs (top-down control), and another mechanism that controls which virus-host pairs occupy these niches (bottom-up control). PMID- 19558512 TI - Short-term microbial and physico-chemical variability in low-temperature hydrothermal fluids near 5 degrees S on the Mid-Atlantic Ridge. AB - This study examines the representativeness of low-temperature hydrothermal fluid samples with respect to their chemical and microbiological characteristics. Within this scope, we investigated short-term temporal chemical and microbial variability of the hydrothermal fluids. For this purpose we collected three fluid samples consecutively from the same spot at the Clueless field near 5 degrees S on the southern Mid-Atlantic Ridge over a period of 50 min. During sampling, the temperature was monitored online. We measured fluid chemical parameters, characterized microbial community compositions and used statistical analyses to determine significant differences between the samples. Overall, the three fluid samples are more closely related to each other than to any other tested habitat. Therefore, on a broad scale, the three collected fluid samples can be regarded as habitat representatives. However, small differences are apparent between all samples. One of the Clueless samples even displayed significant differences (P value < 0.01) to the other two Clueless samples. Our data suggest that the observed variations in fluid chemical and microbial compositions are not reflecting sampling artefacts but are related to short-term fluid variability due to dynamic subseafloor fluid mixing. Recorded temporal changes in fact reflect spatial heterogeneity found in the subsurface as the fluid flows through distinctive pathways. While conservative elements (Cl, Si, Na and K) indicate variable degrees of fluid-seawater mixing, reactive components, including Fe(II), O(2) and H(2)S, show that chemical and microbial reactions within the mixing zone further modify the emanating fluids on short-time scales. Fluids entrain microorganisms, which modify the chemical microenvironment within the subsurface biotopes. This is the first study focusing on short-term microbial variability linked to chemical changes in hydrothermal fluids. PMID- 19558513 TI - Metagenomic approach studying the taxonomic and functional diversity of the bacterial community in a mesotrophic lake (Lac du Bourget--France). AB - The main goals of this work were to identify the metabolic pathways of the bacterial community in a lacustrine ecosystem and to establish links between taxonomic composition and the relative abundances of these metabolic pathways. For this purpose, we analysed a 16S rRNA gene library obtained by gene amplification together with a sequence library of both insert ends on c. 7700 fosmids. Whatever the library used, Actinobacteria was the most abundant bacterial group, followed by Proteobacteria and Bacteroidetes. Specific aquatic clades such as acI and acIV (Actinobacteria) or LD12 and GOBB-C201 (Alphaproteobacteria) were found in both libraries. From comparative analysis of metagenomic libraries, the metagenome of this lake was characterized by overrepresentation of genes involved in the degradation of xenobiotics mainly associated with Alphaproteobacteria. Actinobacteria were mainly related to metabolic pathways involved in nucleotide metabolism, cofactors, vitamins, energy, replication and repair. Betaproteobacteria appeared to be characterized by the presence of numerous genes implicated in environmental information processing (membrane transport and signal transduction) whereas glycan and carbohydrate metabolism pathways were overrepresented in Bacteroidetes. These results prompted us to propose hypotheses on the ecological role of these bacterial classes in lacustrine ecosystems. PMID- 19558515 TI - Ecological limits and diversification rate: alternative paradigms to explain the variation in species richness among clades and regions. AB - Diversification rate is one of the most important metrics in macroecological and macroevolutionary studies. Here I demonstrate that diversification analyses can be misleading when researchers assume that diversity increases unbounded through time, as is typical in molecular phylogenetic studies. If clade diversity is regulated by ecological factors, then species richness may be independent of clade age and it may not be possible to infer the rate at which diversity arose. This has substantial consequences for the interpretation of many studies that have contrasted rates of diversification among clades and regions. Often, it is possible to estimate the total diversification experienced by a clade but not diversification rate itself. I show that the evidence for ecological limits on diversity in higher taxa is widespread. Finally, I explore the implications of ecological limits for a variety of ecological and evolutionary questions that involve inferences about speciation and extinction rates from phylogenetic data. PMID- 19558516 TI - Intima-media thickness and flow-mediated vasodilation in asymptomatic subjects with newly detected severe hypercholesterolemia. AB - BACKGROUND: The aim was to establish the predictors of early functional and structural vascular alterations (using intima-media thickness (IMT)) and flow mediated vasodilation (%FMD) as well as to investigate the interrelationship between IMT and %FMD in asymptomatic, never-treated, severe hypercholesterolemia (HH). METHODS: One hundred and twenty patients with asymptomatic, severe, untreated HH and 100 controls were included. ELISA was used for asymmetric dimethylarginine (ADMA) and cellular adhesion molecules, and fluid chromatography for total homocysteine (tHcy). Hewlett Packard SONOS 5500 with a 7.5 MHz transducer and MedicaSoft IMT software were used for evaluation of %FMD and IMT of common carotid artery (CCA). RESULTS: Patients and controls differ with respect to all tested biomarkers (P < 0.05), except for P-selectin and E-selectin (P < 0.05). %FMD was lower in patients (P < 0.001). The IMT of the CCA was higher in patients (P < 0.001). Inverse correlations were found between %FMD and IMT mean and age, ADMA, Apo-B, Apo-B/Apo-A(1), and tHcy. ADMA was established as the most important factor related to %FMD. Age and Apo-B were established as the most important factors related to IMT mean. An inverse correlation was established between %FMD and IMT mean (r(xy)= 0.546; P < 0.001)). If cases with IMT mean > or =1 were excluded, the correlation weakened. In patients with IMT mean > or =1 mm, the correlation did not change. CONCLUSION: In conclusion, asymptomatic, untreated patients with severe hypercholesterolemia are at high risk of having increased IMT of the CCA, especially if there is endothelial dysfunction, verified by %FMD of the brachial artery. PMID- 19558517 TI - Chronic cigarette smoking affects left and right ventricular long-axis function in healthy young subjects: a Doppler myocardial imaging study. AB - OBJECTIVE: Cigarette smoking is one of the major risk factors for coronary artery disease. However, chronic smoking has additional cardiac adverse effects independent of coronary atherosclerosis. We assessed the possible effects of chronic smoking on left and right ventricular (LV, RV) long-axis function using Doppler myocardial imaging (DMI). METHODS: Forty otherwise healthy smokers (mean age = 26+/-3 years) and 40 age-matched nonsmoking controls enrolled. Standard echocardiography protocol was followed by DMI. Peak systolic (V(S)), early (V(E)) and late diastolic (V(A)) velocities, peak systolic strain (S), and strain rate (SR) were measured from septal, lateral, and RV free walls. RESULTS: The baseline characteristics of two groups were similar. There were significant differences regarding Doppler myocardial velocity, S, and SR indices. Septal V(E) (P < 0.04), S (P < 0.0001) and SR (P < 0.02) were significantly reduced in smokers. For both lateral and RV free wall, V(S) (P < 0.003, P < 0.002, respectively), V(E) (P < 0.0001, P < 0.002, respectively), S (P < 0.0001 for both), and SR (P < 0.0001, P < 0.002, respectively) were significantly reduced in smokers. There were significant correlations between the amount of smoking and septal, lateral, and RV free wall S and SR. CONCLUSIONS: Chronic cigarette smoking causes alterations in long-axis systolic and diastolic functions of right and left ventricles in healthy young subjects. These changes can be accurately detected with Doppler myocardial velocity and SR imaging. PMID- 19558518 TI - Tei index correlates with tissue Doppler parameters and reflects neurohormonal activation in patients with an abnormal transmitral flow pattern. AB - BACKGROUND: Tei index (TI) is a Doppler parameter which reflects combined systolic and diastolic function. We aimed to study the relationship between TI, both traditional and tissue Doppler imaging (TDI) echocardiographic parameters and neurohormonal profile in outpatients with diastolic dysfunction expressed by an abnormal transmitral flow pattern. METHODS AND RESULTS: A total of 67 consecutive outpatients with diastolic dysfunction (abnormal transmitral flow pattern) were studied; all patients underwent clinical evaluation, blood sampling for B-type natriuretic peptide (BNP) plasma assaying, echocardiography for the determination of left ventricular ejection fraction (LVEF), dP/dt, left atrium (LA) dimensions, longitudinal systolic (S) and diastolic wall velocities (E'and A'), TI measured with Doppler echocardiography, and mitral regurgitation (MR) quantified on a semicontinuous scale. TI values were significantly correlated with BNP levels (r = 0.33; P < 0.01), LVEF (r =-0.56; P < 0.001), dP/dt (r = 0.52; P < 0.01), S (r =-0.45; P < 0.001), E'(r =-0.36; P < 0.01), A'(r =-0.27; P < 0.05), LA volume (r = 0.35; P < 0.01), and MR (P for trend < 0.05). In a multivariate regression analysis, TI was an independent predictor of increased BNP levels (beta= 0.32; P < 0.05), even after correction for potential confounders. ROC analysis showed as values of TI >0.59 identified subjects with combined systolic and diastolic dysfunction with a sensitivity of 73.8% and a specificity of 71.4%. CONCLUSIONS: In outpatients with diastolic dysfunction, TI, an easy to perform parameter for global ventricular performance assessment, might be useful in identifying subjects with concomitant systolic impairment and neurohormonal activation. PMID- 19558519 TI - Effect of extreme exercise on myocardial function as assessed by tissue Doppler imaging. AB - BACKGROUND: Response of the human heart to exercise has been studied extensively, but little information is available on the effects of exhaustive exercise on cardiac performance. OBJECTIVES: The aim of this study was to evaluate the effect of severe prolong exercise on both left and right ventricular performance. To maximize the sensitivity of our study we used tissue Doppler imaging. METHODS: Participants in army ranger training program were invited to participate in this prospective study. All patients underwent transthoracic echocardiography using tissue Doppler imaging before and after Ranger training program. RESULTS: A total of 45 consecutive male rangers who completed 8 weeks of training were included in this study. Peak systolic myocardial velocity (S) decreased significantly after training (12.46 +/- 0.54 vs. 9.93 +/- 0.45 cm/s; P < 0.001). In the right ventricle, tissue Doppler measures of systolic and early diastolic function decreased significantly after training compared to pretraining values. CONCLUSION: In conclusion, very strenuous prolonged exercise may result in depressed left ventricular contractile function. This raises the possibility of cardiac fatigue. PMID- 19558520 TI - Evaluation of right ventricular function by using tissue Doppler imaging in patients after repair of tetralogy of fallot. AB - BACKGROUND: The aim of this study was to assess the relation between plasma B type natriuretic peptide (BNP) levels and right ventricular function evaluated by tissue Doppler imaging (TDI) in patients after repair of tetralogy of Fallot (ToF). METHODS: Twenty-five patients with a mean age of 14.1 +/- 4.4 years who underwent repair of ToF at a mean age of 4.9 +/- 5.1 years enrolled in this study. The control group consisted of 29 healthy children at a mean age of 13.1 +/- 2.8 years. The right ventricle and pulmonary regurgitation (PR) were assessed by two-dimensional echocardiography and color Doppler. Blood samples for BNP levels were taken and TDI was performed at rest. RESULTS: Plasma BNP levels were significantly higher in patients than in controls (28.3 +/- 24.1 vs. 7.4 +/- 2.3 pg/mL, P = 0.0001). The myocardial performance index (MPI) (1.08 +/- 0.35 vs. 0.58 +/- 0.11, P = 0.0001) was higher and isovolumic acceleration (IVA) (3.1 +/- 0.7 vs. 5.4 +/- 1.0 m/s(2), P = 0.0001) was lower in patients. The correlations were also significant between the degree of PR and MPI (r = 0.7, P = 0.0001) and also IVA (r =-0.7, P = 0.0001). The correlations were also significant between the BNP level and MPI (r = 0.6, P = 0.0001), IVA (r =-0.4, P = 0.002) and the degree of PR (r = 0.6, P = 0.0001). CONCLUSION: As a result, plasma BNP level increases in patients with ToF and both MPI and IVA from the right ventricular basal segments might be used to assess the right ventricular function. PMID- 19558521 TI - Worsening of left ventricular end-systolic volume and mitral regurgitation without increase in left ventricular dyssynchrony on acute interruption of cardiac resynchronization therapy. AB - BACKGROUND: Responders to cardiac resynchronization therapy (CRT) have greater left ventricular (LV) dyssynchrony than nonresponders prior to CRT. AIM: We conducted this study to see whether the long term responders have more worsening of LV dyssynchrony and LV function on acute interruption of CRT. MATERIALS AND METHODS: We identified 22 responders and 13 nonresponders who received CRT as per standard criteria for 23.73 +/- 7.9 months (median 24.5 months). We assessed the acute change in LV function, mitral regurgitation (MR) and compared LV dyssynchrony in CRT on and off modes. RESULTS: On turning off CRT, there was no significant worsening of LV dyssynchrony in both responders and nonresponders. The dyssynchrony measurements by SPWMD, TDI and 3D echocardiography did not correlate significantly. LVESV increased (p = 0.02) and MR (p = 0.01) worsened in CRT-off mode in responders only without significant change in LVEF or LV dimensions. DISCUSSION AND CONCLUSION: In long-term responders to CRT, there is alteration in the function of remodeled LV with acute interruption of CRT, without significant worsening of LV dyssynchrony. The role of different echocardiographic parameters in the assessment of LV dyssynchrony remains controversial. Even after long-term CRT reversely remodels the LV, the therapy needs to be continued uninterrupted for sustained benefits. PMID- 19558514 TI - Evidence for structuring of bacterial community composition by organic carbon source in temperate lakes. AB - Water entering lakes from the surrounding watershed often delivers large amounts of terrestrial-derived dissolved organic carbon (DOC) that can contribute to aquatic bacterial production. However, research suggests that phytoplankton derived DOC is more labile than its terrestrial counterpart, owing to microbial processing of terrestrial-derived DOC along its flow path to surface waters. The ratio of water colour (absorbance at 440 nm) to chlorophyll a has been suggested as a simple measure of the relative contribution of terrestrial and aquatic primary production to aquatic secondary production. To explore the correlation between primary DOC source and the occurrence of bacterial taxonomic groups, we conducted a survey of bacterial 16S rRNA gene composition in 15 lakes positioned along a water colour : chlorophyll a gradient. Our goal was to identify bacterial taxa occurrence patterns along the colour : chlorophyll a gradient that may indicate a competitive advantage for bacterial taxa using terrestrial or aquatic carbon. We observed a large number of bacterial taxa occurrence patterns suggestive of carbon substrate niche partitioning, especially when relatively highly resolved taxonomic groups were considered. Our survey supports the hypothesis that bacterial taxa partition along a carbon substrate source gradient and highlights carbon source-bacterial interactions that should be explored further. PMID- 19558522 TI - Potential effects of the United States-Mexico border fence on wildlife. AB - Security infrastructure along international boundaries threatens to degrade connectivity for wildlife. To explore potential effects of a fence under construction along the U.S.-Mexico border on wildlife, we assessed movement behavior of two species with different life histories whose regional persistence may depend on transboundary movements. We used radiotelemetry to assess how vegetation and landscape structure affect flight and natal dispersal behaviors of Ferruginous Pygmy-Owls (Glaucidium brasilianum), and satellite telemetry, gene flow estimates, and least-cost path models to assess movement behavior and interpopulation connectivity of desert bighorn sheep (Ovis canadensis mexicana). Flight height of Pygmy-Owls averaged only 1.4 m (SE 0.1) above ground, and only 23% of flights exceeded 4 m. Juvenile Pygmy-Owls dispersed at slower speeds, changed direction more, and had lower colonization success in landscapes with larger vegetation openings or higher levels of disturbance (p < or = 0.047), which suggests large vegetation gaps coupled with tall fences may limit transboundary movements. Female bighorn sheep crossed valleys up to 4.9 km wide, and microsatellite analyses indicated relatively high levels of gene flow and migration (95% CI for F(ST)=0.010-0.115, Nm = 1.9-24.8, M =10.4-15.4) between populations divided by an 11-km valley. Models of gene flow based on regional topography and movement barriers suggested that nine populations of bighorn sheep in northwestern Sonora are linked by dispersal with those in neighboring Arizona. Disruption of transboundary movement corridors by impermeable fencing would isolate some populations on the Arizona side. Connectivity for other species with similar movement abilities and spatial distributions may be affected by border development, yet mitigation strategies could address needs of wildlife and humans. PMID- 19558524 TI - Putting density back into the habitat-quality equation: case study of an open nesting forest bird. AB - Ecological traps and other cases of apparently maladaptive habitat selection cast doubt on the relevance of density as an indicator of habitat quality. Nevertheless, the prevalence of these phenomena remains poorly known, and density may still reflect habitat quality in most systems. We examined the relationship between density and two other parameters of habitat quality in an open-nesting passerine species: the Ovenbird (Seiurus aurocapilla). We hypothesized that the average individual bird makes a good decision when selecting its breeding territory and that territory spacing reflects site productivity or predation risk. Therefore, we predicted that density would be positively correlated with productivity (number of young fledged per unit area). Because individual performance is sensitive to events partly determined by chance, such as nest predation, we further predicted density would be weakly correlated or uncorrelated with the proportion of territories fledging young. We collected data in 23 study sites (25 ha each), 16 of which were located in untreated mature northern hardwood forest and seven in stands partially harvested (treated) 1-7 years prior to the survey. Density explained most of the variability in productivity (R(2)= 0.73), and there was no apparent decoupling between density and productivity in treated plots. In contrast, there was no significant relationship between density and the proportion of territories fledging >or=1 young over the entire breeding season. These results suggest that density reflects habitat quality at the plot scale in this study system. To our knowledge this is one of the few studies testing the value of territory density as an indicator of habitat quality in an open-nesting bird species on the basis of a relatively large number of sizeable study plots. PMID- 19558523 TI - Management decentralization and montane forest conditions in Tanzania. AB - We examined how differences in local forest-management institutions relate to disparate anthropogenic forest disturbance and forest conditions among three neighboring montane forests in Tanzania under centralized, comanaged, or communal management. Institutional differences have been shaped by decentralization reforms. We conducted semistructured interviews with members of forest management committees, local government, and village households and measured anthropogenic disturbance, tree structure, and species composition in forest plots. We assessed differences in governance system components of local institutions, including land tenure, decision-making autonomy by forest users, and official and de facto processes of rule formation, monitoring, and enforcement among the three management strategies. We also assessed differences in frequencies of prohibited logging and subsistence pole cutting, and measures of forest condition. An adjacent research forest served as an ecological reference for comparison of forest conditions. Governance was similar for comanaged and centralized management, whereas communal managers had greater tenure security and decision making autonomy over the use and management of their forest. There was significantly less illegal logging in the communal forest, but subsistence pole cutting was common across all management strategies. The comanaged forest was most disturbed by recent logging and pole cutting, as were peripheral areas of the larger centralized forest. This manifested in more degraded indicators of forest conditions (lower mean tree size, basal area, density of trees >or= 90 cm dbh, and aboveground biomass and higher overall stem density). Greater tenure security and institutional autonomy of the communal strategy contributed to more effective management, less illegal logging, and maintenance of good forest conditions, but generating livelihood benefits was a challenge for both decentralized strategies. Our results underscore the importance of well-designed institutional arrangements in forest management and illustrate mechanisms for improved forest governance and conservation in the context of Tanzanian decentralization reforms. PMID- 19558525 TI - Vaginal and endocervical microorganisms in symptomatic and asymptomatic non pregnant females: risk factors and rates of occurrence. AB - Physiological or non-physiological factors may affect the vaginal flora. The occurrence of genital microorganisms in non-pregnant females of all ages was studied, as were the risk factors associated with each microorganism. A retrospective analysis of vaginal and endocervical cultures and wet smears from 27,172 non-pregnant women, between 1996 to 2005, was performed taking into consideration clinical and socio-demographic characteristics. No microorganisms were observed in 55.7% of the individuals studied and 44.3% had positive cultures. There was no microbiological aetiology in 49% of women with genital symptoms. Poor hygiene, chemical irritants, sexual behaviour, vaginal blood, birth control type, and/or the lack of an oestrogen effect may have caused the symptoms. The highest occurrence of Gram-negative bacteria (p<0.01), mainly Escherichia coli, was observed in prepubescent girls. The highest occurrence of Candida species (p<0.01) was in women of childbearing age, and of Gram-positive bacteria (p<0.01) in menopausal women. Adolescents, particularly asymptomatic girls, carried more frequently Ureaplasma urealyticum and Chlamydia trachomatis (p<0.01). Hormonal contraception and consistent condom use was protective against bacterial vaginosis and U. urealyticum colonization. Users of intrauterine devices had an increased risk of bacterial vaginosis or of contracting U. urealyticum, Mycoplasma hominis and Candida species. Genital complaints were an independent indicator of Candida species, Gram-negative and Gram-positive bacteria, Trichomonas vaginalis and bacterial vaginosis.Chlamydia trachomatis infections were often asymptomatic. It is concluded that the hormonal milieu and non-physiological factors are major determinants of the vaginal flora. If diagnosis of genital infections is based on symptoms alone and not on culture results, it may be erroneous. Sexual abuse should be investigated when a child presents with a sexually transmitted disease. PMID- 19558526 TI - The molecular epidemiology and evolution of the Panton-Valentine leukocidin positive, methicillin-resistant Staphylococcus aureus strain USA300 in Western Australia. AB - Between 2003 and 2008, 76 clinical isolates of the Panton-Valentine leukocidin positive Staphylococcus aureus strain 'West Australian methicillin-resistant Staphylococcus aureus (MRSA)-12' (WA MRSA-12) were recovered from 72 patients living in the Perth area in Western Australia. These isolates were found to belong to multilocus sequence type 8, and had a USA300-like pulsed-field gel electrophoresis pulsotype. All isolates were genotyped using diagnostic DNA arrays covering species markers, resistance factors, virulence-associated, as well as MSCRAMM (microbial surface components recognizing adhesive matrix molecules) genes to prove the identity between WA MRSA-12 and the pandemic strain USA300, as well as to detect possible genetic variability. In general, WA MRSA-12 isolates were similar to USA300, and the most common variant was identical to USA300-TC1516. From this clone, most of the other variants may have evolved by a limited number of gene losses or acquisitions. Variations in carriage of virulence and resistance-associated genes allow distinction of variants or sub clones. Altogether, 16 variants could be distinguished. They differed in the carriage of resistance genes (blaZ/I/R, ermC, msrA + mpbBM, aadD + mupR, aphA3 + sat, tetK, qacC, merA/B/R/T) of beta-haemolysin-converting phages and of enterotoxins (sek + seq, which were deleted in four isolates). Notably, the arginine catabolic mobile element (ACME) was absent in 12 isolates (15.8%). The mercury resistance (mer) operon, which is usually associated with SCCmec type III elements, was found in several ACME-negative isolates. The present study emphasises the importance of genotyping in detecting the introduction and evolution of significant MRSA strains within a community. PMID- 19558527 TI - Hepatic lipase promoter C-480T polymorphism is associated with serum lipids levels, but not subclinical atherosclerosis: the Cardiovascular Risk in Young Finns Study. AB - The common C-480T polymorphism (rs1800588) of the hepatic lipase gene (LIPC) has been associated with high-density lipoprotein (HDL) cholesterol, atherosclerosis, and coronary artery disease. In this study, we examined whether the polymorphism is associated with serum lipid and lipoprotein concentrations, as well as with subclinical atherosclerosis in Young Finns. The participants comprised 2041 men and women (aged 24-39 years) enrolled in the Cardiovascular Risk in Young Finns Study with complete data concerning the rs1800588 polymorphism and serum lipids concentration. All participants underwent an ultrasound examination for brachial artery flow-mediated vasodilatation (FMD) and carotid artery intima-media thickness (IMT) measurement. The marker of arterial elasticity, carotid artery compliance (CAC), was also calculated by means of ultrasound and concomitant brachial blood pressure measurements. In all subjects, serum total cholesterol (p < 0.001), HDL cholesterol (p = 0.006), apolipoprotein AI (apoAI, p < 0.001), and triglyceride (p = 0.009) concentrations increased according to rs1800588 genotype in the order CC, CT, and TT. The same order applied only to apoAI after adjustment for age, body mass index, systolic and diastolic blood pressure, smoking, alcohol consumption, physical activity, diabetes, hypertension, contraceptive hormone use in women, and concentrations of glucose, insulin and C reactive protein in men and women separately (p = 0.007 and p = 0.003, respectively). The polymorphism was also associated with HDL cholesterol, total cholesterol, and triglyceride levels in women (adjusted p = 0.004, p = 0.007 and 0.02, respectively), but not in men (p was not significant for all). No significant association between the rs1800588 and brachial FMD, carotid IMT, or CAC was found among the entire study population or among women or men separately, with or without adjustment for the above-mentioned factors. The rs1800588 is associated with serum lipid and apolipoprotein concentrations, especially in women, but does not seem to be a determinant of brachial artery FMD, carotid IMT, or CAC in young healthy adults. PMID- 19558528 TI - Identification of critical regions for clinical features of distal 10q deletion syndrome. AB - Array comparative genomic hybridization studies were performed to further characterize cytogenetic abnormalities found originally by karyotype and fluorescence in situ hybridization in five clinical cases of distal 10q deletions, including several with complex cytogenetic rearrangements and one with a partial male-to-female sex-reversal phenotype. These results have enabled us to narrow the previously proposed critical regions for the craniofacial, urogenital, and neuropsychiatric disease-related manifestations associated with distal 10q deletion syndrome. Furthermore, we propose that haploinsufficiency of the DOCK1 gene may play a crucial role in the pathogenesis of the 10q deletion syndrome. We hypothesize that alteration of DOCK1 and/or other genes involved in regulation and signaling of multiple pathways can explain the wide range of phenotypic variability between patients with similar or identical cytogenetic abnormalities. PMID- 19558529 TI - Decreased mobilization of endothelial progenitor cells contributes to impaired neovascularization in diabetes. AB - 1. Circulating bone marrow (BM)-derived endothelial progenitor cells (EPCs) play an important role in neovascularization. In the present study, we investigated the mechanisms underlying the reduction in circulating EPCs in a mouse model of diabetes induced by streptozotocin. 2. Compared with non-diabetic controls, diabetic mice had reduced circulating EPCs (0.59 +/- 0.11 vs 0.94 +/- 0.21%, respectively; P < 0.01) and increased plasma endothelial microparticles (18 642 +/- 6809 vs 5692 +/- 1862/mL, respectively; P < 0.01). In a mouse bone marrow (BM) transplantation model, increased adhesion of transplanted BM cells to aortas of diabetic mice was observed compared with control (900 +/- 194 vs 431 +/- 109 cells/mm(2), respectively; P < 0.01). 3. Following hindlimb ischaemia, diabetic mice exhibited suppressed EPC mobilization, a reduction in the expected increase in capillary density and suppressed restoration of transcutaneous oxygen pressure in the ischaemic tissue. Diabetic mice also showed impaired ischaemia-induced upregulation of vascular endothelial growth factor (VEGF), hypoxia-inducible factor (HIF)-1 alpha and interleukin-1 beta, an exaggerated increase in matrix metalloproteinase (MMP)-2 and -9 and a suppressed increase in tissue inhibitor of matrix metalloproteinase (TIMP)-1. On multivariate analysis, VEGF expression was the only independent factor related to circulating EPC count. 4. In conclusion, the data indicate that the decrease in basal circulating EPCs in diabetes may be attributable, in part, to consumptive loss of EPCs due to increased endothelial damage. Impairment of ischaemia-induced EPC mobilization in the diabetic mouse model is associated with altered HIF-1 alpha/VEGF and MMP/TIMP regulation and represents a novel mechanism underlying defective postischaemic neovascularization in diabetes. PMID- 19558530 TI - Secondary pseudotumour cerebri in a patient undergoing sexual reassignment therapy. PMID- 19558531 TI - Chromaticity co-ordinates of Ishihara plates reveal that hidden digit plates can be read by S-cones. AB - BACKGROUND: The Ishihara pseudoisochromatic plates constitute one of the most commonly used screening tools for red-green colour vision deficiencies. Even though hidden digit plates are supposed to be read only by those who are colour vision defective, studies report that some normal trichromats can indeed read these plates. By measuring the chromaticity co-ordinates of the dots used in Ishihara plates, the purpose of this study was to clarify the mechanism that enables normal trichromats and colour vision defectives to read the plates, particularly hidden digit plates. METHODS: Spectrophotometric measurements were made for a 24-plate version of the Ishihara pseudoisochromatic plates and chromaticity co-ordinates of the dots were expressed in the MacLeod-Boynton diagram. RESULTS: As theoretically expected, reading of Ishihara plates by normal trichromats was mediated by the dot chromaticity differences along the L/(L + M) axis. On the other hand, reading by colour vision defective observers was made possible mainly by the dot chromaticity differences along the S/(L + M) axis. This would also explain why some normal trichromats can read hidden digit plates, the plates that are supposed to be read only by colour vision defective observers. CONCLUSION: Normal trichromats read Ishihara plates using their chromatic discrimination ability along the L/(L + M) axis. Red-green colour vision defective observers rely on S-cones in reading the plates. Some normal trichromats can read the hidden digit plates because they can extract S-cone differences efficiently despite the distraction from the L/(L + M) axis. PMID- 19558532 TI - An atypical cutaneous presentation of vasculitis with features of Churg-Strauss syndrome, associated with anti-neutrophil cytoplasmic antibodies and anti glomerular basement membrane antibodies. AB - We report the case of a 59-year-old woman who presented with a persistent papular and nodular cutaneous eruption and new-onset asthma, with normal renal function but persistent haematuria and proteinuria. Investigations revealed eosinophilia, both antineutrophil cytoplasmic antibodies and antiglomerular basement membrane antibodies on serological testing (double-positive vasculitis), and a focal necrotizing glomerulonephritis on renal biopsy. Histological examination of a skin biopsy showed a dense neutrophilic infiltrate with focal fibrinoid necrosis and few eosinophils. The clinical and pathological features suggested a double positive vasculitis/Churg-Strauss overlap syndrome presenting with a predominantly neutrophilic dermatosis. Specific cutaneous features in patients with double-positive vasculitis have not been documented previously. The patient has responded extremely well to immunosuppressive treatment and her disease is currently in remission. PMID- 19558533 TI - Systemic chemerin is related to inflammation rather than obesity in type 2 diabetes. AB - BACKGROUND: The adipokine chemerin modulates the function of innate immune cells and may link obesity and inflammation, and therefore, a possible relation of chemerin to inflammatory proteins in obesity and type 2 diabetes (T2D) was analysed. As visceral fat contributes to systemic inflammation, chemerin was measured in portal venous (PVS), hepatic venous (HVS) and systemic venous (SVS) blood of patients with liver cirrhosis. PATIENTS AND METHODS: Systemic chemerin was determined by ELISA in the serum of normal-weight, overweight and T2D males, in the serum of T2D patients of both sexes, and in PVS, HVS and SVS of patients with liver cirrhosis. RESULTS: Circulating chemerin was similar in T2D and obese individuals but was significantly elevated in both cohorts compared to normal weight individuals. Chemerin positively correlated with leptin, resistin and C reactive protein (CRP). In T2D, chemerin was similar in male and female patients and increased in patients with elevated CRP. Chemerin was similar in PVS and SVS, indicating that visceral fat is not a major site of chemerin synthesis. Higher levels of chemerin in HVS demonstrate that chemerin is also released by the liver. CONCLUSIONS: Visceral fat is not a major site of chemerin release, and elevated systemic levels of chemerin in obesity and T2D seem to be associated with inflammation rather than body mass index. PMID- 19558536 TI - Peri-ictal normalization of visual cortex excitability in migraine: an MEG study. AB - To delineate if the change in cortical excitability persists across migraine attacks, visual evoked magnetic fields (VEF) were measured in patients with migraine without aura during the interictal (n = 26) or peri-ictal (n = 21) periods, and were compared with 30 healthy controls. The visual stimuli were checkerboard reversals with four different check sizes (15', 30', 60' and 120'). For each check size, five sequential blocks of 50 VEF responses were recorded to calculate the percentage change of the P100m amplitude in the second to the fifth blocks in comparison with the first block. At check size 120', interictal patients showed a larger amplitude increment than controls [28.1 +/- 38.3% (s.d.) vs. 8.7 +/- 21.3%] in the second block and a larger increment than peri-ictal patients in the second (28.1 +/- 38.3% vs. -3.2 +/- 19.2%), fourth (22.7 +/- 31.2% vs. -5.7 +/- 22.3%) and fifth (20.5 +/- 30.4% vs. -10.8 +/- 30.1%) blocks (P < 0.05). There was no significant difference at other check sizes or between peri-ictal patients and controls. In conclusion, there may be peri-ictal normalization of visual cortical excitability changes in migraine that is dependent on the spatial frequency of the stimuli and reflects a dynamic modulation of cortical activities. PMID- 19558534 TI - Phenotypic characteristics of familial glucocorticoid deficiency (FGD) type 1 and 2. AB - CONTEXT: Familial glucocorticoid deficiency (FGD) is a rare autosomal recessive disorder as a result of mutation in genes encoding either the ACTH receptor [melanocortin 2 receptor (MC2R)] or its accessory protein [melanocortin 2 receptor accessory protein (MRAP)]. The disorder is known as FGD type 1 and 2, respectively. OBJECTIVE: The aim of the study was to compare the phenotype/genotype relationships between FGD 1 and 2. DESIGN AND PATIENTS: Forty patients with missense MC2R mutations and 22 patients with MRAP mutations were included. Forty-four of these patients had been referred for genetic screening and 18 were patients published by other authors. RESULTS: The median age at presentation for FGD type 1 was variable at 2.0 years; range 0.02-16 years, and this was associated with unusually tall stature, mean height SDS + 1.75 +/- 1.53 (mean +/- SD). In contrast, FGD type 2 presented at a much earlier median age (0.08 years; range at birth to 1.6 years) (P < 0.01) and patients were of normal height SDS + 0.12 +/- 1.35 (P < 0.001). No differences in baseline cortisol or ACTH levels were seen between FGD types 1 and 2. CONCLUSION: FGD type 2 appears to present earlier. This may reflect the functional significance of the underlying mutations in that all MRAP mutations are nonsense or splice site mutations that result in abolition of a functional protein, whereas most of the MC2R mutations are missense mutations and give rise to proteins with some residual function. Tall stature is associated with mutations in MC2R but not in MRAP. There were no other significant clinical distinctions between the two. PMID- 19558535 TI - Intracisternal injection of inflammatory soup activates the trigeminal nerve system. AB - The release of calcitonin gene-related peptide (CGRP) and sensitization of the trigeminal nerve system are important elements in migraine pathophysiology. Sensitization can be induced by topical meningeal administration of inflammatory soup (IS). CGRP release is a marker of trigeminal nerve activation. We examined the effect of intracisternal IS administration on CGRP release in rat jugular vein blood at baseline, 2 and 15 min after the beginning of IS infusion. IS administration caused a significant increase of CGRP levels after 2 and 15 min compared with baseline. Daily oral treatment with topiramate for 4 and 8 weeks led to a dose- and time-dependent reduction of IS-induced CGRP release. Sumatriptan also attenuated stimulated neuropeptide release. These results indicate that intracisternal IS administration leads to activation of the trigeminal system. The inhibition of CGRP release by topiramate offers a possible mechanism that may in part account for the preventative antimigraine activity of this drug. PMID- 19558537 TI - The Tolosa-Hunt syndrome in children: a case report. AB - Tolosa-Hunt syndrome (THS) is characterized by unilateral painful ophthalmoplegia with oculomotor paresis, associated with an idiopathic granulomatous inflammation involving the cavernous sinus, with a typical relapsing-remitting course. We report a case of an 8-year-old girl who was admitted because of an ophthalmoplegia with exotropia and ptosis of the left eyelid, accompanied by diplopia and left sovraorbital pain. The clinical data, neuroradiological findings and response to steroid treatment suggested THS, as defined by the 2004 International Classification of Headache Disorders (ICHD)-II criteria. THS must be considered a possible cause of painful ophthalmoplegia in childhood, as well as in adults, and confirmed with a focused neuroradiological investigation. The few paediatric cases described in the literature that meet the 2004 ICHD-II criteria are not sufficient to identify possible differences between the paediatric and the adult forms. Every new paediatric case should therefore be reported in order to gather and compare further information. PMID- 19558539 TI - Weight change and clinical markers of cardiovascular disease risk during preventive treatment of migraine. AB - Migraine, particularly migraine with aura, and increased body weight are independent risk factors for cardiovascular disease (CVD). The association of weight change and clinical markers of CVD risk was evaluated in subjects participating in a randomized double-blind, parallel-group study of migraine preventive treatment comparing 100 mg/day of topiramate and amitriptyline. Individuals from both treatment groups were pooled and stratified into three groups. The 'major weight gain' group gained > or = 5% of their baseline body weight at the conclusion of the study; the 'major weight loss' group lost > or = 5% of their baseline body weight. The third group had < 5% of weight change. The influence of weight change in headache outcomes, as well as in markers of CVD (blood pressure, cholesterol, C-reactive protein), was assessed using analysis of covariance. Of 331 subjects, 52 (16%) experienced major weight gain and 56 (17%) experienced major weight loss. Weight change was not associated with differential efficacy for the treatment of headache. However, contrasted with those with major weight loss, those who gained weight experienced elevations in mean diastolic blood pressure (+2.5 vs. -1.2 mmHg), heart rate (+7.6 vs. -1.3 beats per minute), glycosylated haemoglobin (+0.09% vs. -0.04%), total cholesterol (+6.4 vs. -6.3 mg/dl), low-density lipoprotein cholesterol (+7.0 vs. -4.4 mg/dl) and triglycerides (+15.3 vs. -10.4 mg/dl) and an increase in high-sensitivity C reactive protein (+1.8 vs. -1.9 mg/l). Both groups experienced decreases in systolic blood pressure (-4.0 vs. -1.3 mmHg) and high-density lipoprotein cholesterol (-3.7 vs. -0.8 mg/dl). Increased weight during migraine treatment is not associated with poor headache treatment outcomes, but is associated with deterioration of CVD risk markers. PMID- 19558538 TI - Genetic association study of endothelin-1 and its receptors EDNRA and EDNRB in migraine with aura. AB - The effect of endothelin-1 and its receptors EDNRA and EDNRB in migraine with aura (MA) susceptibility is not established yet. We studied the association between the MA end-diagnosis and three migraine trait components and 32 single nucleotide polymorphisms (SNPs) capturing the variation of endothelin genes in 850 Finnish migraine patients and 890 non-migrainous individuals. The SNPs showing evidence of association were further studied in 648 German migraine patients and 651 non-migrainous individuals. No significant association was detected. However, the homozygous minor genotype (5% in cases) of the EDNRA SNP rs2048894 showed nominal association with MA both in the Finnish sample (P = 0.015) and in the pooled sample [odds ratio (OR) 1.61, 95% confidence interval (CI) 1.12-2.32, P = 0.010] when adjusted for gender and sample origin. The trait age of onset < 20 years was also associated with rs2048894 (OR 1.69, 95% CI 1.13 2.54, P = 0.011) in the pooled sample. To confirm this finding studies on even larger samples are required. PMID- 19558540 TI - Glossopharyngeal neuralgia triggered by non-noxious stimuli at multiple cephalic and extracephalic sites. AB - Glossopharyngeal neuralgia (GN) triggered by non-noxious stimuli at multiple cephalic and extracephalic sites with positron emission tomography (PET) evidence for involvement of the upper brainstem has never been reported. We present such a patient, a 73-year-old man who since the age of 50 had suffered from GN with a high recurrence rate and very severe unilateral, non-familial GN episodes with very easy trigger zones widely extending beyond the n IX territory. Extensive neuroimaging and neurophysiological tests detected no precise underlying cause. PET scan revealed activation in the upper brainstem on extracephalic triggers. Single-fibre electromyography data will be discussed. We hypothesize that deficient inhibition as seen in trigeminal nociceptive reflexes on the level of brainstem interneurons, a functional lesion in the primary somatosensory cortex sensory thalamic nuclei circuit and the dorsal column-thalamic pathway both activated by light touch may in part be involved in the extracephalic triggering. PMID- 19558541 TI - Changes in functional vasomotor reactivity in migraine with aura. AB - Migraine with aura (MA) is associated with cerebral hyper- and hypoperfusion during and after the attacks. Several attempts to estimate individual perfusion changes and asymmetries in patients with MA using transcranial Doppler (TCD) have not been consistent. In 70 patients with MA and 40 controls with migraine without aura (MoA) or without any history of migraine, interictally recorded TCD sequences were prospectively analysed. Formal curve analysis of the visually evoked flow response (VEFR) was performed semiautomatically. As a main parameter for functional vasomotor reactivity (fVMR), the visually evoked flow rate (VEFR%) was calculated. The VEFR% showed a significantly higher mean difference of 14.7 +/- 12% in MA patients vs. 5.8 +/- 4.4% (P < 0.001) in controls. The significant asymmetry of fVMR in MA patients is suggested to reflect interattack persisting vasomotor changes which are of pathophysiological interest and may be used as a monitoring tool under prophylactic medication. PMID- 19558542 TI - Influence of fibrin glue on seroma formation after modified radical mastectomy: a prospective randomized study. PMID- 19558543 TI - Physiological evidence for interaction between the HIV-1 co-receptor CXCR4 and the cannabinoid system in the brain. AB - BACKGROUND AND PURPOSE: The chemokine, stromal cell-derived growth factor-1alpha (SDF-1alpha/CXCL12), a member of the CXC chemokine family, and the ligand for CXCR4, the co-receptor involved in the entry of human immunodeficiency virus-1 (HIV-1), was tested for its possible interaction with a physiological response to a cannabinoid. EXPERIMENTAL APPROACH: The cannabinoid agonist, an aminoalkylindole, (+)-WIN 55,212-2 [(4,5-dihydro-2-methyl-4(4-morpholinylmethyl) 1-(1-naphthalenyl-carbonyl)-6H-pyrrolo[3,2,1ij]quinolin-6-one], was infused directly into the preoptic anterior hypothalamus (POAH), the primary brain area involved in thermoregulation. KEY RESULTS: WIN 55,212-2 (5-15 microg) evoked a dose-related hypothermia, which was attenuated by SDF-1alpha/CXCL12 microinjected directly into the POAH. The inhibitory effect of SDF-1alpha/CXCL12 on WIN 55,212 2-induced hypothermia was reversed by 1,1'-[1,4 phenylenebis(methylene)]bis[1,4,8,11-tetraazacyclotetradecane] octohydrobromide dihydrate, an antagonist of SDF-1alpha/CXCL12, acting at its receptor, CXCR4. CONCLUSION AND IMPLICATIONS: This study provides the first in vivo evidence for a thermoregulatory interaction between the HIV-1 co-receptor and the cannabinoid system in the brain. PMID- 19558544 TI - Agonist-specific patterns of beta 2-adrenoceptor responses in human airway cells during prolonged exposure. AB - BACKGROUND AND PURPOSE: Beta(2)-adrenoceptor agonists (beta(2)-agonists) are important bronchodilators used in the treatment of asthma and chronic obstructive pulmonary disease. At the molecular level, beta(2)-adrenergic agonist stimulation induces desensitization of the beta(2)-adrenoceptor. In this study, we have examined the relationships between initial effect and subsequent reduction of responsiveness to restimulation for a panel of beta(2)-agonists in cellular and in vitro tissue models. EXPERIMENTAL APPROACH: Beta(2)-adrenoceptor-induced responses and subsequent loss of receptor responsiveness were studied in primary human airway smooth muscle cells and bronchial epithelial cells by measuring cAMP production. Receptor responsiveness was compared at equi-effective concentrations, either after continuous incubation for 24 h or after a 1 h pulse exposure followed by a 23 h washout. Key findings were confirmed in guinea pig tracheal preparations in vitro. KEY RESULTS: There were differences in the reduction of receptor responsiveness in human airway cells and in vitro guinea pig trachea by a panel of beta(2)-agonists. When restimulation occurred immediately after continuous incubation, loss of responsiveness correlated with initial effect for all agonists. After the 1 h pulse exposure, differences between agonists emerged, for example isoprenaline and formoterol induced the least reduction of responsiveness. High lipophilicity was, to some extent, predictive of loss of responsiveness, but other factors appeared to be involved in determining the relationships between effect and subsequent loss of responsiveness for individual agonists. CONCLUSIONS AND IMPLICATIONS: There were clear differences in the ability of different beta(2) agonists to induce loss of receptor responsiveness at equi-effective concentrations. PMID- 19558545 TI - Mammalian P2X7 receptor pharmacology: comparison of recombinant mouse, rat and human P2X7 receptors. AB - BACKGROUND AND PURPOSE: Acute activation of P2X7 receptors rapidly opens a non selective cation channel. Sustained P2X7 receptor activation leads to the formation of cytolytic pores, mediated by downstream recruitment of hemichannels to the cell surface. Species- and single-nucleotide polymorphism-mediated differences in P2X7 receptor activation have been reported that complicate understanding of the physiological role of P2X7 receptors. Studies were conducted to determine pharmacological differences between human, rat and mouse P2X7 receptors. EXPERIMENTAL APPROACH: Receptor-mediated changes in calcium influx and Yo-Pro uptake were compared between recombinant mouse, rat and human P2X7 receptors. For mouse P2X7 receptors, wild-type (BALB/c) and a reported loss of function (C57BL/6) P2X7 receptor were also compared. KEY RESULTS: BzATP [2,3-O-(4 benzoylbenzoyl)-ATP] was more potent than ATP in stimulating calcium influx and Yo-Pro uptake at rat, human, BALB/c and C57BL/6 mouse P2X7 receptors. Two selective P2X7 receptor antagonists, A-740003 and A-438079, potently blocked P2X7 receptor activation across mammalian species. Several reported P2X1 receptor antagonists [e.g. MRS 2159 (4-[(4-formyl-5-hydroxy-6-methyl-3 [(phosphonooxy)methyl}-2-pyridinyl)azo]-benzoic acid), PPNDS and NF279] blocked P2X7 receptors. NF279 fully blocked human P2X7 receptors, but only partially blocked BALB/c P2X7 receptors and was inactive at C57BL/6 P2X7 receptors. CONCLUSIONS AND IMPLICATIONS: These data provide new insights into P2X7 receptor antagonist pharmacology across mammalian species. P2X7 receptor pharmacology in a widely used knockout background mouse strain (C57BL/6) was similar to wild-type mouse P2X7 receptors. Several structurally novel, selective and competitive P2X7 receptor antagonists show less species differences compared with earlier non selective antagonists. PMID- 19558547 TI - (-)-Epigallocatechin gallate suppresses the growth of human hepatocellular carcinoma cells by inhibiting activation of the vascular endothelial growth factor-vascular endothelial growth factor receptor axis. AB - The receptor tyrosine kinase vascular endothelial growth factor (VEGF) receptor (VEGFR) plays an important role in tumor angiogenesis of hepatocellular carcinoma (HCC). (-)-Epigallocatechin gallate (EGCG), the major biologically active component of green tea, inhibits growth in a variety of human cancer cells by inhibiting the activation of several types of receptor tyrosine kinases. In this study, we examined the effects of EGCG on the activity of the VEGF-VEGFR axis in human HCC cells. The levels of total and phosphorylated (i.e. activated) form of VEGFR-2 protein (p-VEGFR-2) were observed to increase in a series of human HCC cell lines in comparison to the Hc normal human hepatocytes. EGCG preferentially inhibited the growth of HuH7 HCC cells, which express constitutive activation of the VEGF-VEGFR axis, in comparison to Hc cells. Treatment of HuH7 cells with EGCG caused a time- and dose-dependent decrease in the expression of VEGFR-2 and p VEGFR-2 proteins. The production of VEGF from HuH7 cells was reduced by treatment with EGCG. Drinking of EGCG significantly inhibited the growth of HuH7 xenografts in nude mice and this was associated with inhibition of the activation of VEGFR-2 and its related downstream signaling molecules, including ERK and Akt. EGCG drinking also decreased the expression of Bcl-x(L) protein and VEGF mRNA in the xenografts. These findings suggest that EGCG can exert, at least in part, its growth-inhibitive effect on HCC cells by inhibiting the VEGF-VEGFR axis. EGCG might therefore be useful in the treatment of HCC. PMID- 19558546 TI - Bovine glycomacropeptide induces cytokine production in human monocytes through the stimulation of the MAPK and the NF-kappaB signal transduction pathways. AB - BACKGROUND AND PURPOSE: Bovine glycomacropeptide (BGMP) is a natural milk peptide that is produced naturally in the gastrointestinal tract during digestion. Glycomacropepide has intestinal anti-inflammatory activity, but the mechanism of action is unknown. Here we have characterized the effects of BGMP on monocytes. EXPERIMENTAL APPROACH: We have used human THP-1 cells as an in vitro monocyte model. The effect of BGMP on the secretion of tumour necrosis factor (TNF), interleukin (IL)-1beta and IL-8 was assessed, as well as the involvement of the NF-kappaB and MAP kinase signalling pathways. The stimulatory effect of BGMP was also tested in human peripheral blood monocytes. KEY RESULTS: BGMP up-regulated the secretion of TNF, IL-1beta and IL-8 in a concentration-dependent fashion. The biological activity was exerted by the intact peptide, because cytokine secretion was not affected by protease inhibitors. The secretion of IL-8 and specially TNF and IL-1beta was blocked by PD98059, SP600125, SB203580 and Bay11-7082, suggesting the involvement of the MAP kinases p38, c-Jun N-terminal kinase and ERK and particularly the NF-kappaB pathway, although IL-8 secretion was independent of p38. BGMP was shown to elicit the phosphorylation of IkappaB-alpha and the nuclear translocation of the NF-kappaB subunits p50 and p65. The effect of BGMP on cytokine secretion was validated in human primary blood monocytes. CONCLUSIONS AND IMPLICATIONS: BGMP stimulates human monocytes, operating via MAP kinase and NF-kappaB pathways. BGMP may exert an indirect intestinal anti inflammatory effect by potentiating host defences against invading microorganisms. PMID- 19558548 TI - Genetic variants of C1orf10 and risk of esophageal squamous cell carcinoma in a Chinese population. AB - Chromosome 1 open reading frame 10 (C1orf10) is either down-regulated or absent in esophageal squamous cell carcinoma (ESCC) tissues compared to its corresponding normal counterparts, and it is involved in heat shock and ethanol response and is expected to protect esophageal epithelium from damage. In the present study, we sequenced DNA samples from 32 individuals to search for genetic variants in the promoter region, coding region, and the untranslated region of C1orf10. Genotypes were analyzed in 991 patients and 984 controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by logistic regression. Luciferase assays were carried out to find the functional SNPs. Six strongly linked single nucleotide polymorphisms (SNPs) spanning a region of 7 kb, 1747G/T, -1139G/C, -1079G/A, -900G/T, Gly480Ser, and 4666G/A were identified (D'= 1, r(2 )= 1). Only one SNP -1139G/C was selected to analyze the association between C1orf10 genotypes and risk of ESCC. Subjects with the -1139CC genotype had a greater risk of developing ESCC compared with those with the -1139GG genotype (adjusted OR = 1.34; 95% CI, 1.02-1.76). There appears to be an interaction between the -1139G/C polymorphism and tobacco smoking that contributes to the risk for ESCC. However, we did not detect any obvious difference in reporter gene assay driven by each allele of C1orf10 promoter or 3' UTR. These data showed that C1orf10 haplotypes containing -1747G/T, -1139G/C, 1079G/A, -900G/T, Gly480Ser, and 4666G/A are significantly associated with susceptibility to ESCC. PMID- 19558550 TI - Subclinical onychomycosis is associated with tinea pedis. AB - BACKGROUND: Onychomycosis is a common cause of nail dystrophy and may be associated with tinea pedis. The presence of dermatophyte fungi in clinically normal nails is unknown. OBJECTIVES: To assess the presence of dermatophyte fungi in normal-appearing toenails and to compare the risk of subclinical dermatophytosis in patients without and with concurrent tinea pedis. METHODS: This is a prospective, University-based study of adults without and with microscopically confirmed tinea pedis. Subjects with dystrophy of any toenail were excluded, as were those ever previously diagnosed as having onychomycosis and those who had used topical antifungals in the past year. A great toenail clipping obtained from each subject was submitted for periodic acid-Schiff histology. RESULTS: One hundred and one subjects (63 men and 38 women, mean +/- SD age 45.4 +/- 15.7 years) were included. Overall, septate hyphae (ostensibly dermatophyte) were identified in seven specimens. Of the 66 control subjects, one case (1.5%) of nail dermatophyte was identified. Of the 35 subjects with tinea pedis, six cases (17%) of nail dermatophyte were identified (P = 0.0066; odds ratio 13.4, 95% confidence interval 1.6-117). There were no significant differences in age or gender between the experimental and control groups or between the nail dermatophyte-positive and negative cohorts. CONCLUSIONS: Dermatophyte fungi may be isolated from normal-appearing toenails. The presence of dermatophytes in this situation is strongly associated with the presence of tinea pedis. Subclinical dermatophyte in the nail plate may serve as a reservoir for ongoing local infection. PMID- 19558549 TI - Antibody-based proteomics for esophageal cancer: Identification of proteins in the nuclear factor-kappaB pathway and mitotic checkpoint. AB - To identify the molecular background of esophageal cancer, we conducted a proteomics study using an antibody microarray consisting of 725 antibodies and surgical specimens from three cases. The microarray analysis identified 24 proteins with aberrant expression in esophageal cancer compared with the corresponding normal mucosa. The overexpression of 14 of the 24 proteins was validated by western blotting analysis of the same samples. These 14 proteins were examined by immunohistochemistry, in which nine proteins showed consistent results with those obtained by western blotting. Among the nine proteins, seven were localized in tumor cells, and two in infiltrating cells. The former included proteins associated with mitotic checkpoint control and the nuclear factor (NF) kappaB pathway. Although mitotic checkpoint gene products (budding uninhibited by benzidazoles 1 homolog beta (BubR1) and mitotic arrest deficient-like 1 (Mad2)) have previously been reported to be involved in esophageal cancer, the association of NF-kappaB-activating kinase, caspase 10, and activator protein-1 with esophageal cancer has not been previously reported. These proteins play a key role in the NF-kappaB pathway, and NF-kappaB is a signal transduction factor that has emerged as an important modulator of altered gene programs and malignant phenotype in the development of cancer. The association of these proteins with esophageal cancer may indicate that mitotic checkpoint gene products and NF kappaB play an important part in the carcinogenesis of esophageal cancer. PMID- 19558551 TI - Chloracne in seven organic chemists exposed to novel polycyclic halogenated chemical compounds (triazoloquinoxalines). AB - Chloracne is an acneiform eruption caused though poisoning by aromatic compounds (usually halogenated) showing a specific molecular configuration. We describe an outbreak of chloracne among seven discovery chemists who synthesized novel polycyclic halogenated chemical compounds which were classified as triazoloquinoxalines, not known to be chloracnegenic. The diagnosis of chloracne, made clinically, elicited a thorough risk assessment and monitoring programme by the occupational health department. The chemists were investigated by serum excretion rates, skin sampling for Propionibacterium acnes, skin biopsy and laboratory blood investigations. Sebum excretion was normal in five cases, raised in one case and severely reduced in another. Skin levels of P. acnes were normal in all patients except for the one subject who had low sebum excretion, in whom they were undetectable. One subject had a slightly raised serum level of alanine aminotransferase. There were no other signs of systemic toxicity. Two subjects were treated with an oral antibiotic, two received topical therapy only and three required no treatment at all. The patients have had thorough health surveillance at 6-monthly and yearly intervals. In each case the chloracne mostly resolved within 18-24 months although on examination about 3 years later, five of the seven still showed minor changes of chloracne. This outbreak emphasizes the need for vigilance in discovery science. The triazoloquinoxalines were not previously recognized as being chloracnegens although their chemical characteristics were subsequently identified as being in keeping with other chemicals that can cause chloracne. Chloracne can be a difficult diagnosis to make when it occurs in a novel setting: occupational physicians and dermatologists need to be vigilant when dealing with unusual eruptions in discovery chemists. PMID- 19558552 TI - Clinical characteristics and course of CD8+ cytotoxic variant of mycosis fungoides: a case series of seven patients. AB - BACKGROUND: Fewer than 5% of cases of mycosis fungoides (MF) present with a cytotoxic/suppressor CD8+ phenotype which, despite immunophenotypic similarities with CD8+ aggressive lymphomas, is regarded as a phenotypic variant of MF. Poikilodermatous MF showing a CD8+ phenotype has been reported to have a nonaggressive clinical behaviour and a good response to psoralen plus ultraviolet A treatment. OBJECTIVES: To perform a retrospective study of CD8+ MF cases diagnosed in the skin lymphoma clinic of Andreas Sygros Hospital. METHODS: We analysed the clinical characteristics, the immunophenotypic and molecular indices, as well as the clinical course of these patients. RESULTS: Seven cases of CD8+ MF (6.5% of all cases of cutaneous T-cell lymphoma) were diagnosed during 2002-2007. One of seven patients had stage IA, five stage IB and one stage IIB disease. Clinical characteristics were variable: four of seven patients presented with poikilodermatous plaques (in one of them lesions of lymphomatoid papulosis with CD8+ phenotype coexisted), one patient with classic MF, one with plantar MF and one with follicular MF. The time period between disease onset and diagnosis was long for most patients (up to 33 years). All patients received the recommended treatment according to TNM staging. Five of seven patients had complete remission, one partial response and one stable disease. CONCLUSIONS: Special clinical characteristics, such as hyperpigmentation and poikiloderma, are often noted in CD8+ MF cases. In our series CD8+ MF presented with a long standing disease and indolent course suggesting that CD8+ cytotoxic immunophenotype may represent a marker of mild biological behaviour. PMID- 19558554 TI - Decreased methionine sulphoxide reductase A expression renders melanocytes more sensitive to oxidative stress: a possible cause for melanocyte loss in vitiligo. AB - BACKGROUND: Methionine is one of the major targets of reactive oxygen species (ROS). It is readily oxidized to methionine-S-sulphoxide and methionine-R sulphoxide, which can be reduced by methionine sulphoxide reductase (MSR) A and B, respectively. MSR represents a unique repair mechanism in the skin antioxidant network. It functions both as a protein repairer and as a ROS scavenger. However, the expression and activity of MSR are significantly reduced in vitiligo. OBJECTIVES: To investigate whether the decreased expression of MSRA is one of the reasons why melanocytes are especially vulnerable to oxidative stress in vitiligo. Methods We downregulated MSRA expression in immortalized human epidermal melanocyte cell line PIG1 by using the short interfering RNA (siRNA) targeted gene silencing method. We checked the changes in MSRA transcript and protein level by using reverse transcriptase-polymerase chain reaction and Western blot, respectively. Then we monitored the viability of MSRA-silenced melanocytes under oxidative stress. All statistical analysis was performed by unpaired two-tailed Student's t-test. RESULTS: The siRNA specific for MSRA successfully suppressed MSRA expression in melanocytes. The lower MSRA expression in melanocytes led to an increased sensitivity to oxidative stress, resulting in more cell death. Furthermore, a remarkable loss of viable cells was found in MSRA silenced melanocytes even in the absence of exogenously added oxidative stress. CONCLUSIONS: MSRA is crucial for melanocytes to fight against oxidative stress in vitiligo. In addition, it is also important for normal cell survival. Any means to enhance MSRA appears to have therapeutic potential for the treatment of vitiligo. PMID- 19558553 TI - Cutaneous adverse effects in patients treated with the multitargeted kinase inhibitors sorafenib and sunitinib. AB - BACKGROUND: The multitargeted kinase inhibitors sorafenib and sunitinib have improved treatment of solid tumours including renal cell carcinoma and hepatocellular carcinoma by offering better clinical responses. However, sorafenib and sunitinib are commonly associated with cutaneous toxicity. OBJECTIVES: We conducted this study to make a clinical assessment of the cutaneous toxicities induced by the oral multitargeted kinase inhibitors sorafenib and sunitinib. METHODS: Retrospectively, we reviewed medical records of patients receiving multitargeted kinase inhibitors, including 109 patients on sorafenib for the treatment of renal cell carcinoma or hepatocellular carcinoma and 119 patients receiving sunitinib for treatment of renal cell carcinoma or a gastrointestinal stromal tumour. Clinical data on cutaneous toxicities were collated. We describe the incidences and intensities of toxicities, and analyse the data statistically. RESULTS: The most common cutaneous toxicity was hand-and foot skin reaction (HFSR). Other cutaneous toxicities included alopecia, stomatitis, skin discoloration (hair or face), subungual splinter haemorrhage, facial swelling, facial erythema and xerosis. HFSR and severe stomatitis required therapy modifications to relieve symptoms, but other cutaneous toxicities did not affect treatment course. HFSR was observed in 48% of patients treated with sorafenib and 36% of those treated with sunitinib. Median time to onset was 18.4 days in patients receiving sorafenib and 32.4 days in those receiving sunitinib. HFSR and stomatitis were early symptoms compared with other cutaneous toxicities. Patients with severe HFSR were likely to develop the symptoms at early phases of therapy. A significant correlation between the severity of HFSR and development of alopecia and stomatitis was found. CONCLUSIONS: Multitargeted kinase inhibitors are associated with a significant risk of various cutaneous adverse events. HFSR is the commonest and most serious cutaneous toxicity in patients treated with these drugs. PMID- 19558555 TI - Malignancies associated with dermatomyositis and polymyositis in Taiwan: a nationwide population-based study. AB - BACKGROUND: Previous studies showed that idiopathic inflammatory myopathies (IIM) carried an increased risk of cancers. However, no large-scale study of IIM has been conducted in the Chinese population. OBJECTIVES: We sought to delineate the association of IIM and various cancer types from a nationwide database in Taiwan. METHODS: We analysed the published national data from records of National Health Insurance claims. Cases of dermatomyositis (DM) and polymyositis (PM) from 2000 to 2005 and cancers registered in the catastrophic illness profile from 1997 to 2006 were collected. A nationally representative cohort of 1,000,000 enrollees was included for comparison. RESULTS: In total, 136 patients (12.8%) among 1059 cases of DM and 46 persons (7.0%) among 661 cases of PM carried internal malignancies. Patients with DM tended to have cancers of nasopharynx, lung and breast. On the other hand, patients with PM tended to have breast, uterine cervix and lung cancers. Compared with the general population, DM gave a 10-fold increased risk for cancers, in which a 66-fold increased risk for nasopharyngeal carcinoma and a 31-fold increased risk for lung cancer were the two most significant. For patients with PM, a 6-fold increased risk for cancer was observed. Juvenile DM had a 16-fold increased risk for haematopoietic or lymphoid malignancy. Two thirds of comorbid malignancies were detected shortly after the diagnoses of IIM, within a mean of 1-2 years. Overall, younger patients with IIM carried the highest risk for malignancies, especially those in their twenties and thirties. CONCLUSIONS: This is the first large-scale study to report the associated malignancies and the cancer risk of IIM in Taiwan. PMID- 19558556 TI - Significant upregulation of antimicrobial peptides and proteins in lichen sclerosus. AB - BACKGROUND: Lichen sclerosus (LS) is a chronic inflammatory T cell-driven sclerotic skin condition in which skin barrier disruption frequently occurs. Inflamed and injured epithelia are a particularly rich source of antimicrobial peptides and proteins (AMPs). OBJECTIVES: We aimed to investigate for the first time the expression pattern of AMPs in lesions of LS as compared with healthy skin. METHODS: Twenty-four women with LS as well as 10 healthy women were included in the study. In order to assess the expression of human beta-defensin (hBD)-1, hBD-2, hBD-3, psoriasin (S100A7), the cathelicidin LL-37 and RNase 7, real-time reverse transcriptase-polymerase chain reaction and immunohistochemistry were performed on skin specimens obtained from lesional and healthy skin of the genital region, respectively. RESULTS: Median hBD-2 mRNA levels observed in LS were significantly higher than in controls (0.15 vs. 0.008; P = 0.0037). Moreover, psoriasin (98.2 vs. 28.1; P = 0.0052) mRNA expression was significantly higher in LS lesions as compared with controls. Significant differences in mRNA expression of hBD-2 and psoriasin were also confirmed by immunohistochemistry. For hBD-1, hBD-3, LL-37 and RNase 7, levels did not differ significantly or were significant only at the gene level but not protein level. CONCLUSIONS: We have demonstrated that hBD-2 and psoriasin expression levels in lesional skin of patients with LS are significantly increased when compared with healthy controls. Whether this observation simply reflects an innate defence response caused by an increased risk of local infection, or whether our data indicate a pathogenetic role of AMPs in LS, will be addressed in future studies. PMID- 19558557 TI - Risk factors for positive findings in patients with high-grade T1 bladder cancer treated with transurethral resection of bladder tumour (TUR) and bacille Calmette Guerin therapy and the decision for a repeat TUR. AB - OBJECTIVE: To determine factors predictive of positive findings at the 3-month follow-up evaluation (after transurethral resection of bladder tumour [TUR] and bacille Calmette-Guerin [BCG] therapy) in patients with initial high-grade (HG)T1 bladder cancer, and to assess the depth of lamina propria (LP) invasion and effectiveness of BCG therapy. PATIENTS AND METHODS: In all, 138 patients with initial HGT1-transitional cell carcinoma (TCC) were prospectively assigned, after TUR + BCG and according to depth of LP invasion, to a postBCG-TUR (T1b) or cystoscopy/cytology (T1a) at 3 months. Any finding at 3 months was considered positive. The predictive value of 11 clinical and pathological variables was assessed by chi-squared, Mann-Whitney U and multivariate logistic regression. RESULTS: Of the 138 patients (14 women, mean age 69 years), 42% had T1a and 58% T1b TCC. Tumour size and carcinoma in situ (CIS) were significantly associated with positive findings and present in 26% (36/138) of the patients. The postBCG TUR (T1b cases), was positive in 31% (25/80), including seven infiltrating tumours. On multivariate analysis, again a tumour size of >3 cm (odds ratio, OR, 7.02) and associated CIS (OR 5.4) were significantly related to a positive postBCG-TUR. A secondary finding was that at 20.3 months; patients with T1a TCC, who did not undergo a repeat TUR, did not have increased progression; only 3% (two of 58) had progressed compared with 21% (17/80) of those with T1b/c TCC (P < 0.002). CONCLUSIONS: In initial HGT1-TCC, tumour size and CIS were predictive factors of positive findings at 3 months after the initial TUR + BCG therapy. Patients with HGT1-TCC invading the LP (T1b TCC) had a seven times higher risk of a positive repeat TUR if the initial tumour was >3 cm and a five-fold increased risk if associated with CIS. The repeat TUR after BCG therapy allowed confirmation of complete resection and pathological evaluation of the BCG response. Although data are still preliminary, the strategy of performing a repeat TUR only in cases with LP involvement, i.e. T1b TCC, did not increase the risk of progression in cases with T1a TCC. PMID- 19558558 TI - Endoluminal isoproterenol reduces renal pelvic pressure during semirigid ureterorenoscopy: a porcine model. AB - OBJECTIVE: To investigate the effects on the pressure-flow relation of renal pelvic pressure during semirigid ureterorenoscopy and endoluminal perfusion of isoproterenol (ISO) 0.1 microg/mL, with emphasis on local effects and cardiovascular side-effects, as topically administered ISO effectively and dose dependently causes relaxation of the upper urinary tract in pigs with no concomitant cardiovascular side-effects. MATERIALS AND METHODS: In anaesthetized female pigs (60 kg), 16 macroscopically normal upper urinary tract systems were subjected to ureterorenoscopy. Via a subcostal incision a 6-F catheter was placed in the renal pelvis for pressure measurements, and a semirigid ureteroscope (7.8 F) was inserted retrogradely in the renal pelvis, through which the pelvis was perfused. The blood pressure and heart rate were recorded. The increase in renal pelvic pressure was examined with increasing flow rates (0, 4, 8, 12, 16, 25 and 33 mL/min) with saline alone or saline + ISO 0.1 microg/mL. Perfusion was initiated on the left side, with randomization for adding ISO or not. Thereafter perfusion was done on the right side as a control in each pig. The surgeons were unaware of whether ISO was added or not. RESULTS: The mean (sd) baseline pelvic pressures in the saline and ISO group were 28 (7.1) and 25 (9.8) mmHg, respectively, with no significant difference (P = 0.079). Endoluminal perfusion with ISO significantly inhibited the pelvic pressure increase to perfusion at all perfusion rates. The pressure-flow relation was linear; the maximum relaxation (27%) was obtained at 4 mL/min, from 52 to 38 mmHg during saline alone and ISO 0.1 microg/mL perfusion, respectively. The mean blood pressure did not change significantly (P = 0.330). The mean (sd) heart rate in the saline and ISO group were 109 (4.5) and 97 (2.1) beats/min, respectively (P < 0.001), i.e. a markedly greater rate in the saline than in the ISO group. CONCLUSION: The pressure-flow relation during semirigid ureterorenoscopy was linear. ISO 0.1 microg/mL in saline significantly reduced the pressure-flow relation during semirigid ureterorenoscopy in this porcine model. ISO might be a potential additive to the irrigation fluid during upper urinary tract endoscopic procedures, minimizing pressure increases due to irrigation and manipulation. PMID- 19558559 TI - Key targets of hormonal treatment of prostate cancer. Part 1: the androgen receptor and steroidogenic pathways. AB - OBJECTIVE: Knowledge of the molecular and cellular changes that occur during the transition of hormone-naive to castration-resistant prostate cancer (CRPC) is increasing rapidly. This might provide a window of opportunity for (future) drug development, and for treating patients with these potential devastating states of disease. The objective of this review is to provide an understanding of the mechanisms that prostate cancer cells use to bypass androgen-deprived conditions. METHODS: We searched PubMed for experimental and clinical studies that describe the molecular changes that lead to CRPC. RESULTS: CRPC remains dependent on a functional androgen receptor (AR), AR-mediated processes, and on the availability of intraprostatic intracellular androgens. CRPCs might acquire different (molecular) mechanisms that enable them to use intracellular androgens more efficiently (AR amplification, AR protein overexpression, AR hypersensitivity), use alternative splice variants of the AR protein to mediate androgen-independent AR functioning, and have altered co-activator and co-repressor gene and protein expression. Furthermore, CRPCs might have the ability to synthesise androgens de novo from available precursors through a renewed and up-regulated synthesis of steroid-hormone converting enzymes. Blocking of enzymes key to de novo androgen synthesis could be an alternative means to treat patients with advanced and/or metastatic disease. CONCLUSION: In CRPC, prostate cancer cells still rely on intracellular androgens and on an active AR for growth and survival. CRPCs have gained mechanisms that enable them to use steroids from the circulation more efficiently through altered gene expression, and through a renewed and up regulated synthesis of steroid hormone-converting enzymes. Additionally, CRPCs might synthesise AR isoforms that enable AR mediated processes independent from available androgens. PMID- 19558561 TI - A mathematical model to evaluate control strategies for mechanical circulatory support. AB - Continuous flow ventricular assist devices (VADs) for mechanical circulatory support (MCS) are generally smaller and believed to be more reliable than pulsatile VADs. However, regarding continuous flow, there are concerns about the decreased pulsatility and ventricular unloading. Moreover, pulsatile VADs offer a wider range in control strategies. For this reason, we used a computer model to evaluate whether pulsatile operation of a continuous flow VAD would be more beneficial than the standard constant pump speed. The computer model describes the left and right ventricle with one-fiber heart contraction models, and the systemic, pulmonary, and coronary circulation with lumped parameter hemodynamical models, while the heart rate is regulated with a baroreflex model. With this computer model, both normal and heart failure hemodynamics were simulated. A HeartMate II left ventricular assist device model was connected to this model, and both constant speed and pulsatile support were simulated. Pulsatile support did not solve the decreased pulsatility issue, but it did improve perfusion (cardiac index and coronary flow) and unloading (stroke work and heart rate) compared with constant speed. Also, pulsatile support would be beneficial for developing control strategies, as it offers more options to adjust assist device settings to the patient's needs. Because the mathematical model used in this study can simulate different assist device settings, it can play a valuable role in developing mechanical circulatory support control strategies. PMID- 19558560 TI - Effects of formaldehyde on cardiovascular system in in situ rat hearts. AB - The aim of the present study was to examine the effects of formaldehyde solution on rat left ventricular function and compare it with those in hypertrophic hearts treated with isoproterenol by pressure-volume measurements with the catheter method. After 20-30 min. of intravenous infusion of 3.7% formaldehyde solution (FA) at 10 MUl (3.7 mg)/kg/min, normal and hypertrophic hearts showed significant decreases in left ventricle end-systolic pressure (ESP), heart rate and cardiac output per minute, indicating an acute pumping failure. Hypertrophic hearts showed significantly smaller ESP, stroke volumes and cardiac output than those in normal hearts. Systolic pressure-volume area at midrange left ventricular volume (PVA(mLVV) : a mechanical work capability index) was significantly smaller than that in normal hearts and per cent of mean PVA(mLVV) versus pre-infusion mean value in hypertrophic hearts was significantly decreased compared to normal hearts 30 min. after FA infusion. The marked decrease in pH, base excess and no changes in PaO2 and PaCO2 suggest metabolic acidosis. The correction of metabolic acidosis with 9% NaHCO3 did not influence on the acute pumping failure, indicating that metabolic acidosis did not cause it. Ultrastructural observations revealed marked dilation of the sarcoplasmic reticulum with intact sarcolemmal membranes and no disintegration of muscle myofibrils. Ryanodine receptors and calcium (Ca2+) pumps (SERCA2A) located in the sarcoplasmic reticulum have major roles in the cytosolic Ca2+ handling. Taken together, acute pumping failure by FA may derive from the impairment of Ca2+ handling in the cardiac excitation contraction coupling. PMID- 19558562 TI - In vivo effects of mesalazine or E. coli Nissle 1917 on microsatellite instability in ulcerative colitis. AB - BACKGROUND: Microsatellite instability (MSI) occurs in chronically inflamed colorectal tissue and may evolve to colitis-associated cancer. In vitro data suggest that mesalazine (5-ASA) improves MSI. AIM: To analyse the changes in MSI in 156 distal colonic biopsies of 39 patients with ulcerative colitis that had been treated within a randomized, double-blind trial comparing 5-ASA with E. coli Nissle (EcN). METHODS: Two biopsies had been collected before and after 1 year of treatment. MSI testing was performed using a panel of eight markers, including 3 dinucleotide and 5 mononucleotide repeats. RESULTS: No MSI was observed with any of the mono-repeats, and among dinucleotide repeats, only D5S346 (maps to APC) and D17S250 (p53) were consistently informative. Overall, 31/156 (20%) biopsies displayed MSI. After 1 year, 3/11 patients displayed MSI improvement [change to microsatellite stability (MSS); 1 on 5-ASA, 2 on EcN] at D5S346 and 4/11 showed MSI worsening (change from MSS to MSI; all 5-ASA). For D17S250, the corresponding data were for 3/9 patients (2 on 5-ASA, 1 on EcN) and 2/9 (both on 5-ASA), respectively. CONCLUSIONS: In the set of biopsies taken from patients treated with 1.5 g 5-ASA for 1 year, there was no improvement in the prevalence of MSI in the distal colon. PMID- 19558563 TI - Systematic review: secondary prevention with band ligation, pharmacotherapy or combination therapy after bleeding from oesophageal varices. AB - BACKGROUND: Variable methods are available for secondary prevention after oesophageal variceal bleeding (EVB). AIM: To compare band ligation (BL), pharmacotherapy (PT) and BL+PT for EVB secondary prevention. METHODS: A systematic search of databases, references and meeting abstracts was conducted for randomized trials of BL, PT or BL+PT. The outcomes were mortality, rebleeding and adverse events. A random-effects model was used for meta-analyses. RESULTS: Twelve trials were included (6 BL vs. PT, 4 BL+PT vs. BL, 2 BL+PT vs. PT). All trials used beta-blockers +/- isosorbide mononitrate (ISMN) as PT. Mortality was not significantly different among trials. Rebleeding was not significantly different for BL vs. PT (RR 1.00, 95% CI 0.73-1.37). BL reduced rebleeding compared with PT for trials with mean beta-blocker dose <80 mg/day (RR 0.67, 95% CI 0.49-0.91). There were nonsignificant differences in rebleeding for BL+PT vs. BL (RR 0.57, 95% CI 0.31-1.08) and BL+PT vs. PT (RR 0.76, 95% CI 0.56-1.03). There was no difference in adverse events between BL vs. PT, but was higher with BL+PT vs. BL. CONCLUSION: Band ligation and PT alone are comparable for secondary prevention of rebleeding after EVB. Further trials with adequate PT dosing are required to determine the efficacy of combination BL+PT therapy. PMID- 19558564 TI - Nutrients, not caloric restriction, extend lifespan in Queensland fruit flies (Bactrocera tryoni). AB - Caloric restriction (CR) has been widely accepted as a mechanism explaining increased lifespan (LS) in organisms subjected to dietary restriction (DR), but recent studies investigating the role of nutrients have challenged the role of CR in extending longevity. Fuelling this debate is the difficulty in experimentally disentangling CR and nutrient effects due to compensatory feeding (CF) behaviour. We quantified CF by measuring the volume of solution imbibed and determined how calories and nutrients influenced LS and fecundity in unmated females of the Queensland fruit fly, Bactocera tryoni (Diptera: Tephritidae). We restricted flies to one of 28 diets varying in carbohydrate:protein (C:P) ratios and concentrations. On imbalanced diets, flies overcame dietary dilutions, consuming similar caloric intakes for most dilutions. The response surface for LS revealed that increasing C:P ratio while keeping calories constant extended LS, with the maximum LS along C:P ratio of 21:1. In general, LS was reduced as caloric intake decreased. Lifetime egg production was maximized at a C:P ratio of 3:1. When given a choice of separate sucrose and yeast solutions, each at one of five concentrations (yielding 25 choice treatments), flies regulated their nutrient intake to match C:P ratio of 3:1. Our results (i) demonstrate that CF can overcome dietary dilutions; (ii) reveal difficulties with methods presenting fixed amounts of liquid diet; (iii) illustrate the need to measure intake to account for CF in DR studies and (iv) highlight nutrients rather than CR as a dominant influence on LS. PMID- 19558565 TI - National and international aspects of doing harm reduction better. PMID- 19558566 TI - Syringe exchange in the United States: doing the simple things better? PMID- 19558567 TI - Harm reduction in the United States at a moment of change: moving innovation from grassroots to mainstream? PMID- 19558568 TI - Expanded access to SEPs and other harm reduction measures in France. PMID- 19558569 TI - Cues must increase smoking behaviour to be clinically relevant. PMID- 19558570 TI - Cue-reactivity in rodents-how predictable? PMID- 19558571 TI - The continuing conundrum of craving. PMID- 19558572 TI - Responses to smoking cues are relevant to smoking and relapse. PMID- 19558573 TI - Immunohistochemical expression of epithelial cell markers in corneas with congenital aniridia and ocular cicatrizing pemphigoid. AB - PURPOSE: We investigated the immunohistochemical characteristics of corneal specimens in congenital aniridia and pemphigoid using various corneal markers to determine the status of the corneal epithelium. METHODS: Conjunctivalization was clinically suspected in all corneas. Ten aniridia and seven pemphigoid paraffin embedded corneal specimens were stained with periodic Schiff reagent (PAS) and antibodies against CK3/12, CK12, CK19, breast cancer resistance protein 1 (BCRP) and p63. RESULTS: Aniridia: six cases contained goblet cells, four were negative. Both groups had cases with (three of six; one of four) and without CK19 positivity and cases with (two of six; three of four) and without p63 positivity. All aniridia cases except two in the goblet cell group were CK3/12- and CK12 positive and BCRP-negative. Pemphigoid: only one of the seven cases contained goblet cells. This case stained positively for CK19, 3/12, 12 and p63 and negatively for BCRP. The other six cases were positive for CK3/12, five of which were positive for CK12; only one case was CK19-positive. Three cases were p63 positive and two BCRP-positive. The CK12 staining was heterogenous in most cases and was often found in the superficial layer. CONCLUSION: Three different stages of epithelial characteristics were found in congenital aniridia and pemphigoid: (i) CK19-negative and inhomogenous CK12-positive cases indicating epithelium mainly from (partly) CK12-deficient limbal stem cells; (ii) CK19- and/or goblet cell-positive and CK12-positive cases with their epithelia originating from CK12 deficient limbal stem cells and from incursing conjunctival cells; and (iii) CK19 positive and CK12-negative cases consisting of conjunctival cells alone. PMID- 19558574 TI - Anterior ciliary artery ligation with simultaneous intravitreal bevacizumab: an adjuvant treatment for end-stage neovascular glaucoma. PMID- 19558575 TI - Intraoperative floppy iris syndrome (IFIS): a practical approach to medical and surgical considerations in cataract extractions. AB - Intraoperative floppy iris syndrome (IFIS) during cataract surgery is characterized by iris fluttering, iris prolapse towards the incisions, and a progressive pupillary constriction leading to high rates of complications. The syndrome has been reported following the treatment of benign prostatic hyperplasia with alpha-1(a) adrenergic receptor inhibitors, especially tamsulosin. The present paper describes the syndrome and discusses its pharmacological background. Several techniques to prevent and to deal with the syndrome are presented. PMID- 19558576 TI - Impact of experimental colitis on hepatobiliary transporter expression and bile duct injury in mice. AB - BACKGROUND AND AIMS: The pathogenetic link between ulcerative colitis and sclerosing cholangitis (SC) is unclear. We hypothesized that colitis induces changes in bile composition via inflammation-induced reduction of hepatobiliary transporter gene expression, ultimately resulting in cholestasis and bile duct injury. METHODS: Alterations in transporter expression and bile secretion in acute dextran sulphate sodium (DSS)-induced colitis were compared with lipopolysaccharide (LPS)-treated mice serving as positive control. Whether chronic DSS-colitis elicits cholangitis in genetically predisposed animals was studied in heterozygous multidrug resistance gene 2 knockout mice (Mdr2(+/-)). RESULTS: LPS but not DSS-colitis changed major hepatobiliary transporters (Ntcp, Bsep, Mrp2-4, Ostalpha/beta, Abcg5/8, Oatp1-4, Mdr1b and Mdr2), enzymes (Cyp3a11 and Cyp7a1), nuclear receptors (RXRalpha, FXR, CAR and PXR) and proinflammatory mediators (tumour necrosis factor alpha and inducible nitric oxide synthase). Formation of toxic bile reflected by an increased bile acid/phospholipid ratio was observed neither in acute nor in chronic colitis, although heterozygous Mdr2(+/-) mice developed mild portal inflammation after chronic colitis. CONCLUSIONS: In contrast to LPS, DSS-colitis has a minor impact on hepatobiliary gene expression and bile secretion. Therefore, intestinal inflammation-associated changes of hepatobiliary transporter expression do not play a pathogenetic role in SC. PMID- 19558577 TI - New curcumin analogues exhibit enhanced growth-suppressive activity and inhibit AKT and signal transducer and activator of transcription 3 phosphorylation in breast and prostate cancer cells. AB - Curcumin, the active component of turmeric, has been shown to protect against carcinogenesis and prevent tumor development in cancer. To enhance its potency, we tested the efficacy of synthetic curcumin analogues, known as FLLL11 and FLLL12, in cancer cells. We examined the impact of FLLL11 and FLLL12 on cell viability in eight different breast and prostate cancer cell lines. FLLL11 and FLLL12 (IC(50) values 0.3-5.7 and 0.3-3.8 micromol/L, respectively) were substantially more potent than curcumin (IC(50) values between 14.4-50 micromol/L). FLLL11 and FLLL12 were also found to inhibit AKT phosphorylation and downregulate the expression of HER2/neu. In addition, we demonstrate for the first time that FLLL11 and FLLL12 inhibit phosphorylation of signal transducer and activator of transcription (STAT) 3, an oncogene frequently found to be persistently active in many cancer types. The inhibition of STAT3 signaling was confirmed by the inhibition of STAT3 DNA binding and STAT3 transcriptional activity. Furthermore, FLLL11 and FLLL12 were more effective than curcumin in inhibiting cell migration and colony formation in soft agar as well as inducing apoptosis in cancer cells. These results indicate that FLLL11 and FLLL12 exhibit more potent activities than curcumin on the inhibition of STAT3, AKT, and HER 2/neu, as well as inhibit cancer cell growth and migration, and may thus have translational potential as chemopreventive or therapeutic agents for breast and prostate cancers. PMID- 19558579 TI - Evaluating minimal important differences for the FACT-Melanoma quality of life questionnaire. AB - OBJECTIVES: Minimal Important Differences (MIDs) establish benchmarks for interpreting mean differences in clinical trials involving quality of life outcomes and inform discussions of clinically meaningful change in patient status. The purpose of this study was to assess MIDs for the Functional Assessment of Cancer Therapy-Melanoma (FACT-M). METHODS: A prospective validation study of the FACT-M was performed with 273 patients with stages I through IV melanoma. FACT-M, Karnofsky Performance Scales, and Eastern Cooperative Oncology Group Performance Status scores were obtained at baseline and 3 months following enrollment. Anchor- and distribution-based methods for assessing MIDs were compared, and pattern-mixture modeling was employed to derive multivariate adjusted estimates. RESULTS: This study indicates that an approximate range for MIDs of the FACT-M subscales is between 5 to 9 points for the Trial Outcome Index, 4 to 6 points for the Melanoma Combined Subscale, 2 to 4 points for the Melanoma Subscale, and 1 to 2 points for the Melanoma Surgery Subscale. Each method produced similar but not identical ranges of MIDs. CONCLUSIONS: The properties of the anchor instrument employed to derive MIDs directly affect resulting MID ranges and point values. When MIDs are offered as supportive evidence of a clinically meaningful change, the anchor instrument used to derive clinically meaningful thresholds of change should be clearly stated along with information supporting the choice of anchor instrument as the most appropriate for the domain of interest. PMID- 19558578 TI - Selective NTPDase2 expression modulates in vivo rat glioma growth. AB - The ectonucleoside triphosphate diphosphohydrolases (E-NTPDases) are a family of ectoenzymes that hydrolyze extracellular nucleotides, thereby modulating purinergic signaling. Gliomas have low expression of all E-NTPDases, particularly NTPDase2, when compared to astrocytes in culture. Nucleotides induce glioma proliferation and ATP, although potentially neurotoxic, does not evoke cytotoxic action on the majority of glioma cultures. We have previously shown that the co injection of apyrase with gliomas decreases glioma progression. Here, we tested whether selective re-establishment of NTPDase2 expression would affect glioma growth. NTPDase2 overexpression in C6 glioma cells had no effect on in vitro proliferation but dramatically increased tumor growth and malignant characteristics in vivo. Additionally, a sizable platelet sequestration in the tumor area and an increase in CD31 or platelet/endothelial cell adhesion molecule 1 (PECAM-1), vascular endothelial growth factor and OX-42 immunostaining were observed in C6-Enhanced Yellow Fluorescent Protein (EYFP)/NTPDase2-derived gliomas when compared to controls. Treatment with clopidogrel, a P2Y(12) antagonist with anti-platelet properties, decreased these parameters to control levels. These data suggest that the ADP derived from NTPDase2 activity stimulates platelet migration to the tumor area and that NTPDase2, by regulating angiogenesis and inflammation, seems to play an important role in tumor progression. In conclusion, our results point to the involvement of purinergic signaling in glioma progression. PMID- 19558580 TI - Spontaneous thrombosis of a large vein of galen malformation. AB - A large vein of Galen was diagnosed in a 9-month-old boy. This was not treated at birth, as there was no associated congestive heart failure. The patient was followed conservatively and follow-up magnetic resonance imaging showed increase in the size of the vein of Galen malformation. Subsequent cerebral angiogram demonstrated hypertrophied but thrombosed right posterior choroidal artery, suggesting spontaneous thrombosis of the arterial feeder and thus the embolization was not pursued. PMID- 19558581 TI - Brainstem involvement in Sjogren's syndrome-related sensory neuronopathy. AB - BACKGROUND AND PURPOSE: Sensory neuronopathies (SN) are peripheral nervous system disorders associated with degeneration of dorsal root ganglion neurons. Magnetic resonance imaging (MRI) studies have shown abnormalities limited to T2-weighted high signal intensity in the posterior columns. METHODS AND RESULTS: A 65-year old woman with Sjogren syndrome had slowly progressive unsteadiness of gait and limb paresthesias. A blink reflex examination suggested a paramedian brainstem lesion, confirmed by MRI. CONCLUSIONS: Sjogren's syndrome-related SN may be associated with a more diffuse immune-mediated aggression, involving also the brainstem, and leading to some of the blink reflex abnormalities observed in nonparaneoplastic SN. PMID- 19558582 TI - Secrets to developing Wnt-age melanoma revealed. PMID- 19558583 TI - Misinformation about Pneumocystis. PMID- 19558584 TI - A comprehensive review of biomarkers in psoriasis. AB - Psoriasis is a common, chronic skin disorder, the pathogenesis of which is incompletely understood. Results from various clinical and experimental studies indicate that psoriasis is a complex, multifactorial disease with a genetic predisposition. Factors such as climate, physical trauma, drug, stress and infections (Streptococcus, human immunodeficiency virus) are known to trigger psoriasis. The success of treatment of psoriasis with T-cell depletion and antitumour necrosis factor (TNF)-alpha treatment is explained by the involvement of T cells and TNF- alpha in the pathogenesis of psoriasis. The biochemical basis for the pathogenesis of psoriasis can be attributed to both overexpression and underexpression of certain proteins in psoriatic lesions. The anomalies in protein expression can be classified as abnormal keratinocyte differentiation, keratinocyte hyperproliferation and inflammation. Oxidative stress (OS) and increased free-radical generation have been linked to skin inflammation in psoriasis. The review presents evidence for various markers of psoriasis that can be targeted for effective treatment, including biomarkers of inflammation, keratinocyte hyperproliferation and abnormal differentiation, and stress. PMID- 19558585 TI - Poisoning deaths among Finnish children from 1969 to 2003. AB - AIM: Trends in paediatric deaths due to poisoning are little studied. The aim of this study was to investigate the cause and secular trend of poisoning deaths among Finnish children. METHODS: Death certificates of all Finnish children aged 0-15 who died due to poisoning between 1969 and 2003 were obtained from the Statistics of Finland and analysed. RESULTS: During the study period, altogether 121 children aged 0-15 years died from poisoning in Finland. Among 0- to 4-year olds, the incidence of poisoning deaths declined to practically zero by the beginning of 1980s. Most of these deaths were unintentional poisonings. Among 5- to 15-year olds, the incidence of poisoning deaths varied during the study period. In this age group, up to 53% of the deaths were suicides among girls compared with 20% among boys (p = 0.017). The corresponding figures for substance abuse were 54% among boys and 9% among girls (p < 0.001). CONCLUSION: Despite the declining secular trend seen in paediatric poisoning deaths in Finland from 1969 to 2003, the risk of death from both intentional and unintentional poisoning persists in children. Health programmes should be continued especially to promote well-being in families and to prevent teenage suicides and substance abuse. PMID- 19558586 TI - Identification of key proteins involved in the anammox reaction. AB - Bacteria performing anaerobic ammonium oxidation (anammox) are key players in the global nitrogen cycle due to their inherent ability to convert biologically available nitrogen to N(2). Anammox is increasingly being exploited during wastewater treatment worldwide, and about 50% of the total N(2) production in marine environments is estimated to proceed by the anammox pathway. To fully understand the microbial functionality and mechanisms that control environmental feedbacks of the anammox reaction, key proteins involved in the reaction must be identified. In this study we have utilized an analytical protocol that facilitates detection of proteins associated with the anammoxosome, an intracellular membrane compartment within the anammox bacterium. The protocol enabled us to identify several key proteins of the anammox reaction including a hydrazine hydrolase producing hydrazine, a hydrazine-oxidizing enzyme converting hydrazine to N(2) and a membrane-bound ATP synthase generating ATP from the gradients of protons formed in the anammox reaction. We also performed immunogold labelling electron microscopy to determine the subcellular location of the hydrazine hydrolase. The results from our study support the hypothesis that proteins associated with the anammoxosome host the complete suite of reactions during anammox. PMID- 19558587 TI - The evaluation of novel chromogenic substrates for the detection of lipolytic activity in clinical isolates of Staphylococcus aureus and MRSA from two European study groups. AB - Eight novel chromogenic substrates were evaluated for their efficacy in detecting lipase activity in clinical isolates of Staphylococcus aureus from the United Kingdom and Malta. All isolates metabolized the chromogenic lipase substrates 5 (4-hydroxy-3,5-dimethoxyphenylmethylene)-2-thioxothia-zolidin-4-one-3-ethanoic acid (SRA)-propionate, SRA-butyrate, SRA-octanoate and 2-[2-(4-hydroxy-3,5 dimethoxyphenyl)-vinyl]-3-methy-benzothiazolium salt (SB(Z)TM)-acetate. Over 90% of the isolates metabolized the lipase substrates SRA-decanoate and SRA-laurate. However, only 0.6% of UK isolates and 2% of Maltese isolates metabolized the lipase substrate SRA-myristate; none of the isolates tested metabolized SB(Z)TM butyrate. Traditional Tween 80 assays showed that over 73% of the UK methicillin resistant Staphylococcus aureus (MRSA) isolates and 83% of the UK methicillin sensitive Staphylococcus aureus (MSSA) isolates demonstrated lipolytic activity. In contrast, Maltese isolates showed lipase activity in 94% and 88% of the MRSA and MSSA strains, respectively. Lipases in MRSA and MSSA demonstrated substrate specificity whose activity appeared dependent upon hydrocarbon chain length of the chromogen. These novel chromogens can be used for lipase enzyme detection and have application for full characterization of numerous S. aureus lipases. PMID- 19558588 TI - Characterization of HetN, a protein involved in heterocyst differentiation in the cyanobacterium Anabaena sp. strain PCC 7120. AB - HetN, a putative ketoacyl reductase, is required for heterocyst pattern maintenance in the filamentous cyanobacterium Anabaena sp. strain PCC 7120. The hetN gene, when present on a multicopy plasmid, is able to suppress heterocyst differentiation. Little is known about the biochemical properties of HetN. In the present study, we found that HetN could hydrolyze ATP or GTP in vitro, and that this activity was dependent on the presence of magnesium in the reaction mixture. Mutations of the conserved active Ser(142)-Tyr(155)-Lys(159) triad, predicted as necessary for the reductase activity of HetN, had only weak effect on the hydrolysis of ATP. The residue Lys(159) is shown to be necessary for the heterocyst-suppressing activity of HetN, as the corresponding mutant allele present on a replicative plasmid failed to block heterocyst differentiation in contrast to the wild type. This result suggests that the reductase activity of HetN is involved in the HetN-mediated inhibition of heterocyst formation. PMID- 19558589 TI - Spatial distribution of sporocarps of stipitate hydnoid fungi and their belowground mycelium. AB - Interest in stipitate hydnoid fungi of the genera Bankera, Hydnellum, Phellodon and Sarcodon has increased due to the decline in numbers of sporocarps in Europe. Conservation of these fungi is hindered by a lack of understanding of their basic ecology. In particular, a better understanding of their belowground ecology is required. Real-time PCR in conjunction with spatially explicit sampling was used to quantify the relationship between sporocarps and mycelium of Hydnellum peckii and Phellodon tomentosus. Species-specific DNA of the target species was quantified in 100 soil samples collected on a 360 x 360 cm grid at five locations where sporocarps were present. All sporocarps within the grid and up to 2 m around the grid were mapped. Sporocarp production did not occur over the whole extent of the belowground mycelium of these two species, and mycelium extended up to 330 cm away from the immediate site of sporocarp production. Spatial analyses using Kernel-smoothing and Moran's I correlograms showed that, with a single exception, there was no quantitative relationship between sporocarp distribution and the belowground abundance of mycelium. These findings have important implications for the conservation of this rare group of fungi. PMID- 19558590 TI - Some highlights of research on aging with invertebrates, 2009. AB - This annual review focuses on invertebrate model organisms, which shed light on new mechanisms in aging and provide excellent systems for both genome-wide and in depth analysis. This year, protein interaction networks have been used in a new bioinformatic approach to identify novel genes that extend replicative lifespan in yeast. In an extended approach, using a new, human protein interaction network, information from the invertebrates was used to identify new, candidate genes for lifespan extension and their orthologues were validated in the nematode Caenorhabditis elegans. Chemosensation of diffusible substances from bacteria has been shown to limit lifespan in C. elegans, while a systematic study of the different methods used to implement dietary restriction in the worm has shown that they involve mechanisms that are partially distinct and partially overlapping, providing important clarification for addressing whether or not they are conserved in other organisms. A new theoretical model for the evolution of rejuvenating cell division has shown that asymmetrical division for either cell size or for damaged cell constituents results in increased fitness for most realistic levels of cellular protein damage. Work on aging-related disease has both refined our understanding of the mechanisms underlying one route to the development of Parkinson's disease and has revealed that in worms, as in mice, dietary restriction is protective against cellular proteotoxicity. Two systematic studies genetically manipulating the superoxide dismutases of C. elegans support the idea that damage from superoxide plays little or no role in aging in this organism, and have prompted discussion of other kinds of damage and other kinds of mechanisms for producing aging-related decline in function. PMID- 19558591 TI - The endoscopic spectrum of segmental colitis associated with diverticulosis. AB - OBJECTIVE: An endoscopic classification of 'Segmental colitis associated with diverticulosis' (SCAD) is lacking. Our aim was therefore to assess the endoscopic spectrum of SCAD, comparing it with the histological and clinical features. METHOD: A prospective study was performed from January 2004 to October 2007. Diagnosis of SCAD was made on the basis of specific endoscopic and histological patterns. RESULTS: A total of 6230 consecutive colonoscopies were performed during the study period. SCAD was diagnosed in 92 (1.48%) patients, with four endoscopic patterns: pattern A, 'crescentic fold disease' (52.20%); pattern B, 'Mild-to moderate ulcerative colitis-like' pattern (30.40%); pattern C, 'Crohn's disease colitis-like' pattern (10.90%); pattern D, 'Severe ulcerative colitis like' pattern (6.50%). Most patients with patterns A (58.33%, P < 0.018) and B (89.29%, P < 0.00001) showed histological alterations resembling moderate ulcerative colitis (UC). In pattern C, larger histological variability was found (P < 0.01). All patients showing pattern D showed the typical histological alteration changes of severe UC (P < 0.0001). In pattern A (60.42%, P = n.s.) and pattern B (46.43%, P = n.s.), diarrhoea was the most common symptom whilst abdominal pain was the most frequent in pattern C (50%, P = n.s.) and pattern D (83.33%, P = n.s.) patients. CONCLUSIONS: Endoscopic patterns of SCAD may range from mild to severe inflammation. The histopathological findings but not clinical features showed a statistically significant association with the degree of endoscopic severity. PMID- 19558592 TI - Mesocolic plane surgery: an old but forgotten technique? PMID- 19558593 TI - DSM-V and the definitions: time to get it right. PMID- 19558594 TI - Immunolocalization of Epstein-Barr virus-related antigens in a case of sweat gland adenocarcinoma. PMID- 19558596 TI - Optimizing technique in elliptical excisional surgery: some pearls for practice. PMID- 19558595 TI - Mutation in DSG1 causing autosomal dominant striate palmoplantar keratoderma. PMID- 19558597 TI - Evaluation of a polymerase chain reaction-restriction fragment length polymorphism assay for dermatophyte and nondermatophyte identification in onychomycosis. AB - BACKGROUND: Dermatophytes are the main cause of onychomycoses, but various nondermatophyte filamentous fungi are often isolated from abnormal nails. The correct identification of the aetiological agent of nail infections is necessary in order to recommend appropriate treatment. OBJECTIVE: To evaluate a rapid polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay based on 28S rDNA for fungal identification in nails on a large number of samples in comparison with cultures. METHODS: Infectious fungi were analysed using PCR-RFLP in 410 nail samples in which fungal elements were observed in situ by direct mycological examination (positive samples). The results were compared with those previously obtained by culture of fungi on Sabouraud agar from the same nail samples. RESULTS: PCR-RFLP identification of fungi in nails allowed validation of the results obtained in culture when Trichophyton spp. grew from infected samples. In addition, nondermatophyte filamentous fungi could be identified with certainty as the infectious agents in onychomycosis, and discriminated from dermatophytes as well as from transient contaminants. The specificity of the culture results relative to PCR-RFLP appeared to be 81%, 71%, 52% and 63% when Fusarium spp., Scopulariopsis brevicaulis, Aspergillus spp. and Candida spp., respectively, grew on Sabouraud agar. It was also possible to identify the infectious agent when direct nail mycological examination showed fungal elements, but negative results were obtained from fungal culture. CONCLUSIONS: Improved sensitivity for the detection of fungi in nails was obtained using the PCR-RFLP assay. Rapid and reliable molecular identification of the infectious fungus can be used routinely and presents several important advantages compared with culture in expediting the choice of appropriate antifungal therapy. PMID- 19558598 TI - Cellular and molecular features of lipoma tissue: comparison with normal adipose tissue. AB - BACKGROUND: Involvement of adipose-derived stem/progenitor/stromal cells (ASCs) in the development of lipomas has been suggested, but the pathogenesis and pathophysiology of this tumour remain unclear. OBJECTIVES: To analyse cellular and transcriptional characteristics of lipoma tissue compared with normal adipose tissue, further to delineate differentiating features. METHODS: For lipoma or normal adipose tissues, we used a new whole-mount staining enabling three dimensional imaging of nonfixed and nonfrozen adipose tissue. Immunohistochemistry and real-time polymerase chain reaction for obesity-related genes were performed as well as comparative assay of the proliferative and adipogenic capacity of ASCs. RESULTS: A large number of small adipocytes surrounded by CD34+/lectin- ASCs and increased numbers of Ki67+/CD34+ ASCs indicated enhanced adipogenesis in lipoma compared with normal adipose tissue. In contrast, cellular apoptosis was not enhanced in lipoma, suggesting that the enlargement of lipoma tissue may be due to a positive balance of adipocyte turnover (accelerated adipogenesis combined with nonenhanced apoptosis). Leptin mRNA was upregulated in lipoma, while adiponectin, tumour necrosis factor-alpha and glucose transporter 1 mRNA were downregulated and there were no apparent changes in hypoxia-inducible factor 1alpha, peroxisome proliferator-activated receptor-gamma and plasminogen activator inhibitor-1. These results suggested dysfunction of lipoma adipocytes similar to that in obesity, but indicated that lipoma tissue lacked several obesity-related phenomena such as ischaemia (hypoxia), macrophage infiltration, inflammatory reactions and enhanced glycolysis. ASCs from lipoma and normal adipose tissue showed similar proliferative and adipogenic capacity. CONCLUSIONS: Our findings revealed that lipoma tissue shows a positive balance of adipocyte turnover involving proliferating ASCs and several transcriptional differences from adipose tissue enlargement in obesity. PMID- 19558599 TI - Temporary brittle bone disease: fractures in medical care. AB - Temporary brittle bone disease is the name given to a syndrome first reported in 1990, in which fractures occur in infants in the first year of life. The fractures include rib fractures and metaphyseal fractures which are mostly asymptomatic. The radiological features of this disorder mimic those often ascribed to typical non-accidental injury. The subject has been controversial, some authors suggesting that the disorder does not exist. This study reports five infants with typical features of temporary brittle bone disease in whom all or most of the fractures took place while in hospital. A non-accidental cause can be eliminated with some confidence, and these cases provide evidence in support of the existence of temporary brittle bone disease. PMID- 19558600 TI - Variations of Apgar score of very low birth weight infants in different neonatal intensive care units. AB - OBJECTIVE: The Apgar score should be an objective method to assess the state of newborns; however, its applicability in preterm infants is hampered by large variations among different observers. The study tested whether physicians that give low scores to written case descriptions also apply lower scores to preterm infants. PATIENTS AND METHODS: Descriptions (BMJ 2004; 329: 143-4) were sent to 14 neonatal units. Physicians were asked to evaluate the Apgar (case score). From seven units Apgar scores of all very low birth weight infants (VLBW) born between January 2004 and December 2006 were obtained from charts (clinical score). RESULTS: In total, 121 physicians from 14 institutions (median 9, range 3-15) replied: 24 residents with <6-month and 28 with >6-month neonatal experience, and 69 consultants. The assessment of the case scores was very heterogeneous with large variations in respiration, muscle tone and reflexes. Clinical scores were obtained from 1000 VLBW infants. The score depended on the gestational age, with a median of 4 at 24 and 7 at 27 weeks. With one exception, centres that assigned low case scores had also low clinical scores. CONCLUSION: There is considerable variation in assigning Apgar scores. Definitions are required to apply the Apgar score to infants under clinical conditions such as preterm delivery, resuscitation or artificial ventilation. PMID- 19558601 TI - Neurotransmitter release: a docking role for UNC-13 proteins (Commentary on Siksou et al.). PMID- 19558602 TI - The GluR2 subunit inhibits proliferation by inactivating Src-MAPK signalling and induces apoptosis by means of caspase 3/6-dependent activation in glioma cells. AB - Glioblastoma multiforme (GBM) is the most invasive and undifferentiated type of brain tumour, and so surgical interventions are ineffective. We found that GluR2 is absent in fast-growing GBM-derived tumour stem cells and high-grade glioma specimens, but is expressed in slow-growing stem cells and low-grade glioma specimens. More remarkably, GluR2 overexpression in U-87MG cells inhibits proliferation by inactivating extracellular signal-regulated kinase (ERK)1/2-Src phosphorylation and induces apoptosis. Mechanistically, we observed that the scaffold protein GRIP is essential for the effect of GluR2 on ERK-Src inactivation. These findings indicate that the absence of the GluR2 subunit favours malignancy. PMID- 19558604 TI - Perceptual influence on bimanual coordination: an fMRI study. AB - In bimanual coordination subjects typically show a spontaneous preference for movement symmetry. While there is experimental evidence for the principle of muscle homology, recent evidence suggested that bimanual coordination may be mediated as perceptual goals (Mechsner et al., 2001). To explore this controversy we performed a fMRI study in 11 healthy, right-handed subjects using bimanual index finger abductions and adductions in a congruous condition, i.e. both palms down, and incongruous conditions with either the left or right palm up. Our fMRI data showed a widespread bihemispheric network mediating proprioceptive coordination of the two hands with significant differences mainly for a perceptual dissociation: in the incongruous conditions with the one palm up there was a BOLD signal increase in a bilateral frontoparietal network involving the motor and premotor cortical areas, particularly in the right palm-up condition. These results accord with the notion that perceptual cues play an important role in the control of bilateral hand movements. PMID- 19558605 TI - Neuropathic pain in spinal cord injury: significance of clinical and electrophysiological measures. AB - A large percentage of spinal cord-injured subjects suffer from neuropathic pain below the level of the lesion (bNP). The neural mechanisms underlying this condition are not clear. The aim of this study was to elucidate the general effects of spinal deafferentiation and of bNP on electroencephalographic (EEG) activity. In addition, the relationship between the presence of bNP and impaired function of the spinothalamic tract was studied. Measurements were performed in complete and incomplete spinal cord-injured subjects with and without bNP as well as in a healthy control group. Spinothalamic tract function, assessed by contact heat evoked potentials, did not differ between subjects with and without bNP; nevertheless, it was impaired in 94% of subjects suffering from bNP. In the EEG recordings, the degree of deafferentiation was reflected in a slowing of EEG peak frequency in the 6-12-Hz band. Taking into account this unspecific effect, spinal cord-injured subjects with bNP showed significantly slower EEG activity than subjects without bNP. A discrimination analysis in the subjects with spinothalamic tract dysfunction correctly classified 84% of subjects as belonging to either the group with bNP or the group without bNP, according to their EEG peak frequency. These findings could be helpful for both the development of an objective diagnosis of bNP and for testing the effectiveness of new therapeutic agents. PMID- 19558603 TI - Attentional modulation of adaptation in V4. AB - Adaptation and visual attention are two processes that alter neural responses to luminance contrast. Rapid contrast adaptation changes response size and dynamics at all stages of visual processing, while visual attention has been shown to modulate both contrast gain and response gain in macaque extrastriate visual cortex. Because attention aims to enhance behaviorally relevant sensory responses while adaptation acts to attenuate neural activity, the question we asked is, how does attention alter adaptation? We present here single-unit recordings from V4 of two rhesus macaques performing a cued target detection task. The study was designed to characterize the effects of attention on the size and dynamics of a sequence of responses produced by a series of flashed oriented gratings parametric in luminance contrast. We found that the effect of attention on the response dynamics of V4 neurons is inconsistent with a mechanism that only alters the effective stimulus contrast, or only rescales the gain of the response. Instead, the action of attention modifies contrast gain early in the task, and modifies both response gain and contrast gain later in the task. We also show that responses to attended stimuli are more closely locked to the stimulus cycle than unattended responses, and that attended responses show less of the phase lag produced by adaptation than unattended responses. The phase advance generated by attention of the adapted responses suggests that the attentional gain control operates in some ways like a contrast gain control utilizing a neural measure of contrast to influence dynamics. PMID- 19558606 TI - Lineage analysis of quiescent regenerative stem cells in the adult brain by genetic labelling reveals spatially restricted neurogenic niches in the olfactory bulb. AB - The subventricular zone (SVZ) of the lateral ventricles is the major neurogenic region in the adult mammalian brain, harbouring neural stem cells within defined niches. The identity of these stem cells and the factors regulating their fate are poorly understood. We have genetically mapped a population of Nestin expressing cells during postnatal development to study their potential and fate in vivo. Taking advantage of the recombination characteristics of a nestin::CreER(T2) allele, we followed a subpopulation of neural stem cells and traced their fate in a largely unrecombined neurogenic niche. Perinatal nestin::CreER(T2)-expressing cells give rise to multiple glial cell types and neurons, as well as to stem cells of the adult SVZ. In the adult SVZ nestin::CreER(T2)-expressing neural stem cells give rise to several neuronal subtypes in the olfactory bulb (OB). We addressed whether the same population of neural stem cells play a role in SVZ regeneration. Following anti-mitotic treatment to eliminate rapidly dividing progenitors, relatively quiescent nestin::CreER(T2)-targeted cells are spared and contribute to SVZ regeneration, generating new proliferating precursors and neuroblasts. Finally, we have identified neurogenic progenitors clustered in ependymal-like niches within the rostral migratory stream (RMS) of the OB. These OB-RMS progenitors generate neuroblasts that, upon transplantation, graft, migrate and differentiate into granule and glomerular neurons. In summary, using conditional lineage tracing we have identified neonatal cells that are the source of neurogenic and regenerative neural stem cells in the adult SVZ and occupy a novel neurogenic niche in the OB. PMID- 19558607 TI - Noise-induced increase in human auditory evoked neuromagnetic fields. AB - Noise is usually detrimental to auditory perception. However, recent psychophysical studies have shown that low levels of broadband noise may improve signal detection. Here, we measured auditory evoked fields (AEFs) while participants listened passively to low-pitched and high-pitched tones (Experiment 1) or complex sounds that included a tuned or a mistuned component that yielded the perception of concurrent sound objects (Experiment 2). In both experiments, stimuli were embedded in low or intermediate levels of Gaussian noise or presented without background noise. For each participant, the AEFs were modeled with a pair of dipoles in the superior temporal plane, and the effects of noise were examined on the resulting source waveforms. In both experiments, the N1m was larger when the stimuli were embedded in low background noise than in the no noise control condition. Complex sounds with a mistuned component generated an object-related negativity that was larger in the low-noise condition. The results show that low-level background noise facilitates AEFs associated with sound onset and can be beneficial for sorting out concurrent sound objects. We suggest that noise-induced increases in transient evoked responses may be mediated via efferent feedback connections between the auditory cortex and lower auditory centers. PMID- 19558608 TI - Development and content validity of a gastroparesis cardinal symptom index daily diary. AB - BACKGROUND: The Gastroparesis Cardinal Symptom Index (GCSI) is a patient-reported outcome for gastroparesis using a two-week recall period. To minimize potential patient recall effects, a daily diary version of the GCSI (GCSI-DD) was developed. AIMS: To evaluate the content validity of GCSI-DD for the symptoms in patients with documented gastroparesis, to capture symptom variability over time and to compare responses of this GCSI-DD to the original GCSI. METHODS: In gastroparesis adults with delayed gastric emptying, cognitive debriefing interviews were conducted to elicit their perspective on relevant symptoms of gastroparesis and relevant recall periods and to evaluate patient understanding of GCSI-DD. Patients completed the GCSI-DD daily over a 2-week period and completed the GCSI at baseline and the 2-week follow-up visit. RESULTS: Twelve gastroparesis patients, of whom five were diabetic and nine women, reported nausea (100%), vomiting (100%), stomach fullness (75%), bloating (58%) and loss of appetite (50%) were important symptoms. All patients understood diary instructions and item content and reported that the diary captured their gastroparesis symptom experience; 83% considered response scales adequate. There was significant daily variability in GCSI-DD scores. Mean GCSI-DD subscale and total scores over 2 weeks correlated strongly (all r > 0.90) with GCSI scores at 2-week follow-up. CONCLUSIONS: The GCSI-DD includes symptoms relevant to patients with gastroparesis, captures daily variability of those symptoms and has psychometric properties consistent with a good patient-reported outcome endpoint for gastroparesis clinical trials. PMID- 19558609 TI - Systematic review: standard- and double-dose proton pump inhibitors for the healing of severe erosive oesophagitis -- a mixed treatment comparison of randomized controlled trials. AB - BACKGROUND: No randomized controlled trial (RCT) has compared all European licensed standard- and double-dose PPIs for the healing of severe erosive oesophagitis. AIM: To compare the effectiveness of licensed doses of PPIs for healing severe erosive oesophagitis (i.e. esomeprazole 40 mg, lansoprazole 30 mg, omeprazole 20 mg and 40 mg, pantoprazole 40 mg and rabeprazole 20 mg). METHODS: Systematic review of CENTRAL, EMBASE and MEDLINE for RCTs in patients with erosive oesophagitis (completed October 2008). Endoscopically verified healing rates at 4 and 8 weeks were extracted and re-calculated if not analysed by intention-to-treat. A mixed treatment comparison was used to combine direct treatment comparisons with indirect trial evidence while maintaining randomization. Odds ratios (OR) are reported compared to omeprazole 20 mg. RESULTS: A total of 3021 papers were identified in the literature search; 12 were of sufficient quality to be included in the analysis. Insufficient data were available to included rabeprazole. Esomeprazole 40 mg was found to provide significantly higher healing rates at 4 weeks [OR 1.84, 95% Credible Interval (95% CrI): 1.50 to 2.22] and 8 weeks (OR 1.91, 95% CrI: 1.13 to 2.88). No other PPI investigated had significantly higher healing rates than omeprazole 20 mg. CONCLUSION: Esomeprazole 40 mg consistently demonstrates higher healing rates compared with licensed standard- and double-dose PPIs. PMID- 19558610 TI - Effect of adjunct metformin treatment on levels of plasma lipids in patients with type 1 diabetes. AB - BACKGROUND: In addition to its glucose-lowering effect, metformin treatment has been suggested to improve lipidaemia in patients with type 2 diabetes. In contrast, in patients with type 1 diabetes (T1DM), information about the effect of metformin treatment on lipidaemia is limited. In this study, we report the effect of a 1-year treatment with metformin vs. placebo on plasma lipids in T1DM patients and persistent poor glycaemic control. METHODS: One hundred T1DM patients with haemoglobinA(1c) (HbA(1c)) > or =8.5% during the year before enrolment entered a 1-month run-in period on placebo treatment. Thereafter, patients were randomized (baseline) to treatment with either metformin (1000 mg twice daily) or placebo for 12 months (double masked). Patients continued ongoing insulin therapy and their usual outpatient clinical care. Outcomes were assessed at baseline and after 1 year. RESULTS: After 1 year, in those patients who did not start or stop statin therapy during the trial, metformin treatment significantly reduced total and LDL cholesterol by approximately 0.3 mmol/l compared with placebo (p = 0.021 and p = 0.018 respectively). Adjustment for statin use or known cardiovascular disease did not change conclusions. In statin users (metformin: n = 22, placebo: n = 13), metformin significantly lowered levels of LDL and non-HDL cholesterol by approximately 0.5 mmol/l compared with placebo (adjusted for changes in statin dose or agent: p = 0.048 and p = 0.033 respectively). HbA(1c) (previously reported) was not significant different between treatments. CONCLUSION: In patients with poorly controlled T1DM, at similar glycaemic levels, adjunct metformin therapy during 1 year significantly lowered levels of proatherogenic cholesterolaemia independent of statin therapy. PMID- 19558611 TI - An NMR-based metabonomic investigation on effects of supplementation with isosteviol or soy protein to diabetic KKAy mice. PMID- 19558613 TI - Predator-prey relationships in a changing environment: the case of the sparrowhawk and its avian prey community in a rural area. AB - 1. Changes in community composition are expected to entail cascading effects at different trophic levels within a food web. However, empirical evidence on the impact of changes in prey communities on the population dynamics of generalist predators, and on the extent of possible feedback processes, remains scarce. 2. We analysed the dynamics of a generalist predator, the European sparrowhawk Accipiter nisus L., in a rural area of Northern Denmark. Over a 20-year period, the diet of the predator has been thoroughly assessed (>30,000 identified prey items) and quantitative information about its avian prey community, based on standard breeding bird surveys, has revealed significant trends for several passerine species, although the overall prey biomass available remained stable. 3. The growth rate of the sparrowhawk breeding population was negatively related to the previous sparrowhawk density and to winter temperature, but was positively related to available prey biomass. Contrary to expectations for a generalist predator, sparrowhawks seemed to be predominantly sensitive to changes in the cumulative abundance of their two main prey species, the skylark Alauda arvensis L. and the blackbird Turdus merula L., but less so to changes in the wider prey community. 4. In demographic terms, the two-phase sparrowhawk dynamic recorded here (a recovery following an initial decrease) was mainly driven by recruitment of yearling females into the breeding population rather than by variation in the apparent survival of breeding females. 5. Our findings emphasize that changes in the composition of a prey community, affected by environmental changes, impacted population dynamics of a generalist predator. Finally, we found conditions that might enable apparent competition between blackbirds and song thrushes Turdus philomelos L. to occur. High blackbird abundance, maintaining sparrowhawks at a relatively high density may, in turn, push song thrushes into a predator pit. PMID- 19558612 TI - Heterogeneous selection on a heritable temperament trait in a variable environment. AB - 1. Temperament traits increasingly provide a focus for investigating the evolutionary ecology of behavioural variation. Here, we examine the underlying causes and selective consequences of individual variation in the temperament trait 'exploration behaviour in a novel environment' (EB, based on an 8-min assay) in a free-ranging population of a passerine bird, the great tit Parus major. 2. First, we conducted a quantitative genetic analysis on EB using a restricted maximum likelihood-based animal model with a long-term pedigree. Although repeatability was relatively high, EB was only moderately heritable and permanent environment (V(PE)) effects contributed as much to phenotypic variance as additive genetic effects. 3. We then asked whether heterogeneous selection acted on EB at various temporal and spatial scales. Using estimates of lifetime reproductive success, we found evidence of weak negative directional selection acting on EB amongst females which was driven by selection through recruitment, but not fecundity, in one of the four breeding years. There was no evidence of any selection on EB through survival. 4. Heterogeneous selection on EB within seasons was also observed amongst males through fecundity along two fine-scale environmental gradients--local breeding density and habitat quality; we are unaware of any previous equivalent demonstrations. 5. All of these analyses were repeated on a second measure of exploration behaviour (EB(2), measured during a 2 min assay) to facilitate comparison with other studies. EB and EB(2) were strongly correlated to one another at the genetic level, but were only moderately correlated at the phenotypic level and V(PE) was undetected in EB(2). Selection on EB(2) was similar to that on EB; we conclude that both traits are broadly equivalent from an evolutionary perspective. 6. Our analyses suggest that to the extent that the temperament trait 'exploration behaviour' is subject to natural selection in this population, this selection is highly context dependent and most evident along two environmental gradients. Furthermore, the strong V(PE) effect detected suggests that understanding the causes and consequences of variation in this trait will require studies firmly embedded in an environmental context. PMID- 19558614 TI - Anesthetic management of a neonate with type IV laryngotracheo-esophageal cleft. AB - Laryngotracheo-esophageal cleft is a rare congenital anomaly that occurs when the trachea and esophagus fail to separate during fetal development. Clinical severity varies greatly in anatomic cleft extent. We report the successful management of the airway of a neonate with type IV laryngotracheo-esophageal cleft for the gastric division surgery by dividing the esophagogastric continuity between the esophageal orifice and the stomach using the balloon catheter and remaining spontaneous breathing through the large cuffed tracheal tube inserted into the esophageal orifice. PMID- 19558615 TI - Data sharing for pharmacokinetic studies. AB - Pooling data from different pediatric studies can provide a single robust pharmacokinetic analysis that allows covariate analysis and hypothesis testing. Data sharing should be driven by the altruistic purpose of improving drug understanding to the clinical benefit of children. Electronic communications have rendered the sharing of data relatively easy, and data sharing within the wider scientific community has become commonplace. Data sharing allows verification of results, save costs and time, allows new interpretation of old data, and can fulfill teaching benefits. It may stimulate cooperative competition between researchers and allow individual researchers to concentrate on unique aspects of the scientific puzzle. However, there is occasionally a reluctance to share, in part because of fear of others stealing the hard work of a research group, which may not be recognized in subsequent publications that reuse data. Providing data may require additional effort for presentation in a suitable format. Data may be abused or used for purposes other than those for which they were collected. Propriety claims may limit access to industry-sponsored drug research. The question of who has ownership of data is contentious. Investigators often consider data they have collected to be their own property. Reputations and grants may be hinge on ownership of a data set. However, other team members, institutions, funding agencies, and the public also have a stake. The difficulties identified in the general scientific community also apply to data sharing for pediatric pharmacokinetic studies. There are few clearly established rules at present, and consideration of the issues hinges on ethical and philosophical arguments. The development of databases will depend on collaboration and cooperation and greater clarity and consensus over appropriate processes and procedures. PMID- 19558616 TI - Who should obtain written consent for magnetic resonance imaging under general anesthesia? AB - Consent for magnetic resonance imaging under anesthesia in children is complex - it does not fit the usual model of consent for invasive procedures and requires the collaboration of multiple specialists from different disciplines. This article discusses the issues surrounding consent for this procedure, sets out four essential elements of the consent process, and proposes that, of the specialists involved, the referring clinician is best placed to discuss the options with parents and obtain written consent. PMID- 19558617 TI - Control of mucosal polymicrobial populations by innate immunity. AB - The gastrointestinal tract carries out the complex process of localizing the polymicrobial populations of the indigenous microbiota to the lumenal side of the GI mucosa while absorbing nutrients from the lumen and preventing damage to the mucosa. This process is accomplished through a combination of physical, innate and adaptive host defences and a 'strategic alliance' with members of the microbiota. To cope with the constant exposure to a diverse microbial community, the GI tract, through the actions of a number of specialized cells in the epithelium and lamina propria, has layers of humoral, physical and cellular defences that limit attachment, invasion and dissemination of the indigenous microbiota. However, the role of the microbiota in this dynamic balance is vital and serves as another level of 'innate' defence. We are just beginning to understand how bacterial metabolites aid in the control of potential pathogens within the microbiota and limit inflammatory responses to the microbiota, concepts that will impact our understanding of the biological effects of antibiotics, diet and probiotics on mucosal inflammatory responses. PMID- 19558618 TI - Neurofibromatosis type 1: should we screen for other genetic syndromes? A case report of co-existence with multiple endocrine neoplasia 2A. AB - BACKGROUND: NF 1 is a genetic disorder with an autosomal dominant pattern of inheritence. It is associated with neoplastic disorders mainly derived from the neural seath. However, the co-existence of NF1 with the full spectrum of MEN 2A has rarely been reported. The aim of the study was to investigate the presence of secondary neoplasias in a patient with diagnosed NF1, and in particular the presence of hyperparathyroidism and the possible co-existence with another pheochromocytoma-related syndrome. METHODS: We report a case of a 70 years old female patient who had NF1. The patient was referred to our center and was diagnosed with an isolated pheochromocytoma of the right adrenal gland for which she underwent right adrenalectomy. We further investigated for the presence of another pheochromocytoma-related syndrome and in particular for the presence of hyperparathyroidism and medullary thyroid cancer. Molecular screening for germline mutations of the genes NF1, RET and VHL has also been performed. RESULTS: The patient was further diagnosed with hyperparathyroidism and medullary thyroid cancer, having the full spectrum of the clinical picture of the MEN2A syndrome. The genetic testing revealed the germline mutation for NF1 but not for the RET proto-oncogene which is generally found in MEN2A cases. CONCLUSION: To our knowledge this is a rare case of co-existence of two pheochromocytoma-related genetic syndromes, and generates the question of whether all patients with these syndromes should undergo a thorough clinical and laboratory investigation for the possibility of another co-existing pheochromocytoma-related genetic syndrome. PMID- 19558619 TI - A common molecular basis for membrane docking and functional priming of synaptic vesicles. AB - Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes execute synaptic vesicle (SV) fusion. Vesicle fusion is preceded by an obligatory Munc13-dependent priming process that conveys fusion competence to SVs by facilitating SNARE complex assembly. Ultrastructural studies after chemical fixation indicated that vesicle docking to the plasma membrane is independent of Munc13s but these results may be misleading because aldehyde fixatives modify the localization of SVs with respect to the plasma membrane. To reinvestigate the role of Munc13s in vesicle docking, cultured hippocampal slices were immobilized using high-pressure freezing, which circumvents aldehyde artifacts. High-pressure freezing was combined with electron tomography to reach a resolution that allows the characterization of details of SV docking in a close-to-native state. In control slices, docked vesicles are not hemifused with the plasma membrane but linked to it and to dense material at the active zone by small strands. In slice cultures from Munc13-deficient mice, vesicles are not docked to the active zone plasma membrane. These results indicate that SV docking at the plasma membrane and functional priming are respective morphological and physiological manifestations of the same molecular process mediated by SNARE complexes and Munc13s. PMID- 19558620 TI - Background luminance affects the detection of microampere currents delivered to macaque striate cortex. AB - Monkeys detect electrical microstimulation delivered to the striate cortex (area V1). We examined whether the ability of monkeys to detect such stimulation is affected by background luminance. While remaining fixated on a spot of light centered on a monitor, a monkey was required to detect a 100 ms train of electrical stimulation delivered to a site within area V1 situated from 1 to 1.5 mm below the cortical surface. A monkey signaled the delivery of stimulation by depressing a lever after which it was rewarded with a drop of apple juice. Control trials were interleaved during which time no stimulation was delivered and the monkey was rewarded for not depressing the lever. Biphasic pulses were delivered at 200 Hz and the current ranged from 2 to 30 microA using 0.2 ms anode first biphasic pulses. The background luminance level of the monitor could be varied from 0.005 to 148 cd/m(2). It was found that, for monitor luminance levels below 10 cd/m(2), the current threshold to evoke a detection response increased. We discuss the significance of this result with regard to phosphenes elicited from human V1 and in relation to visual perception. PMID- 19558621 TI - Wide-ranging frequency preferences of auditory midbrain neurons: Roles of membrane time constant and synaptic properties. AB - Periodicity is a fundamental sound attribute. Its coding has been the subject of intensive research, most of which has focused on investigating how the periodicity of sounds is processed through the synaptic machinery in the brain. The extent to which the intrinsic properties of cells play in periodicity coding, particularly in the creation of selectivity to periodic signals, is not well understood. We performed in vitro whole-cell patch recordings in the frog torus semicircularis to investigate each neuron's intrinsic membrane properties as well as responses to sinusoidal current injected through the electrode and periodic stimulation of the ascending afferent. We found that: (i) toral neurons were heterogeneous, showing diverse biophysical phenotypes having distinct membrane characteristics, including membrane time constants (tau) and ionic channel compositions (I(h), I(kir), I(kv) and I(NaP)); (ii) a neuron's tau was tightly correlated with its current-evoked frequency preference (FP; range: 0.05-50 Hz); (iii) application of blockers for I(h), I(kir) and I(kv) (but not I(NaP)) shifted the tau as well as the cell's current-evoked FP, suggesting that these ion channels contribute to the cell's FP through regulation of tau; (iv) a neuron's tau was also correlated with its afferent-evoked FP (range: 10-300 pulses/s); and (v) the range of afferent-evoked FP was approximately one order higher than the range of current-evoked FPs, suggesting that both the cell's intrinsic membrane and synaptic properties contribute to determining the afferent-evoked cell specific FP (whose range matched those of cell-specific responses to sound stimulation, e.g. selectivity to amplitude modulation rate). PMID- 19558622 TI - Myzus persicae (green peach aphid) salivary components induce defence responses in Arabidopsis thaliana. AB - Myzus persicae (green peach aphid) feeding on Arabidopsis thaliana induces a defence response, quantified as reduced aphid progeny production, in infested leaves but not in other parts of the plant. Similarly, infiltration of aphid saliva into Arabidopsis leaves causes only a local increase in aphid resistance. Further characterization of the defence-eliciting salivary components indicates that Arabidopsis recognizes a proteinaceous elicitor with a size between 3 and 10 kD. Genetic analysis using well-characterized Arabidopsis mutants shows that saliva-induced resistance against M. persicae is independent of the known defence signalling pathways involving salicylic acid, jasmonate and ethylene. Among 78 Arabidopsis genes that were induced by aphid saliva infiltration, 52 had been identified previously as aphid-induced, but few are responsive to the well-known plant defence signalling molecules salicylic acid and jasmonate. Quantitative PCR analyses confirm expression of saliva-induced genes. In particular, expression of a set of O-methyltransferases, which may be involved in the synthesis of aphid repellent glucosinolates, was significantly up-regulated by both M. persicae feeding and treatment with aphid saliva. However, this did not correlate with increased production of 4-methoxyindol-3-ylmethylglucosinolate, suggesting that aphid salivary components trigger an Arabidopsis defence response that is independent of this aphid-deterrent glucosinolate. PMID- 19558623 TI - Moderate heat stress reduces the pH component of the transthylakoid proton motive force in light-adapted, intact tobacco leaves. AB - We measured the DeltaPsi and DeltapH components of the transthylakoid proton motive force (pmf) in light-adapted, intact tobacco leaves in response to moderate heat. The DeltaPsi causes an electrochromic shift (ECS) in carotenoid absorbance spectra. The light-dark difference spectrum has a peak at 518 nm and the two components of the pmf were separated by following the ECS for 25 s after turning the light off. The ECS signal was deconvoluted by subtracting the effects of zeaxanthin formation (peak at 505 nm) and the qE-related absorbance changes (peak at 535 nm) from a signal measured at 520 nm. Heat reduced DeltapH while DeltaPsi slightly increased. Elevated temperature accelerated ECS decay kinetics likely reflecting heat-induced increases in proton conductance and ion movement. Energy-dependent quenching (qE) was reduced by heat. However, the reduction of qE was less than expected given the loss of DeltapH. Zeaxanthin did not increase with heat in light-adapted leaves but it was higher than would be predicted given the reduced DeltapH found at high temperature. The results indicate that moderate heat stress can have very large effects on thylakoid reactions. PMID- 19558624 TI - Does greater leaf-level photosynthesis explain the larger solar energy conversion efficiency of Miscanthus relative to switchgrass? AB - C(4) perennial grasses are being considered for bioenergy because of their high productivity and low inputs. In side-by-side replicated trials, Miscanthus (Miscanthus x giganteus) has previously been found more than twice as productive as switchgrass (Panicum virgatum). The hypothesis that this difference is attributable to higher leaf photosynthetic rates was tested on established plots of switchgrass and Miscanthus in central Illinois with >3300 individual measurements on 20 dates across the 2005 and 2006 growing seasons. Seasonally integrated leaf-level photosynthesis was 33% higher in Miscanthus than switchgrass (P < 0.0001). This increase in carbon assimilation comes at the expense of additional transpiration since stomatal conductance was on average 25% higher in Miscanthus (P < 0.0001). Whole-chain electron transport rate, measured simultaneously by modulated chlorophyll fluorescence, was similarly 23% higher in Miscanthus (P < 0.0001). Efficiencies of light energy transduction into whole chain photosynthetic electron transport, leaf nitrogen use and leaf water use were all significantly higher in Miscanthus. These may all contribute to its higher photosynthetic rates, and in turn, productivity. Systematic measurement of photosynthesis over two complete growing seasons in the field provides a unique dataset explaining why the productivity of these two species differs and for validating mechanistic production models for these emerging bioenergy crops. PMID- 19558626 TI - No evidence of intrauterine transmission of hepatitis A virus from a mother to a premature infant. AB - AIM: To determine whether or not an intrauterine transmission of hepatitis A virus (HAV) occurred from an infected mother to her premature infant delivered by caesarean section. METHODS: The mother and her child were tested for HAV by serology and reverse transcription PCR. RESULTS: An outbreak of HAV infection was seen among children and a 33-year-old day-care teacher, pregnant in third trimester, at a day-care centre in southern Sweden. Due to premature labour and diminished foetal movements a caesarean section was performed and a premature girl in gestational weeks 33 + 1 was born. During the 3-week postnatal hospitalization period the child presented no clinical symptoms of HAV infection and anti-HAV IgM antibodies remained undetectable at day 14 and 109 after birth. Furthermore HAV RNA remained undetectable by reverse transcription PCR in the child's blood at birth and in throat and faeces for the first 3 and 4 weeks of life respectively. HAV RNA in the mother's blood was detected at 6 days prior to and at 17 days after delivery. HAV RNA was undetectable in breast milk when tested on day 3 after delivery. CONCLUSION: There was no evidence of intrauterine transmission of hepatitis A virus from a viraemic mother to her premature child delivered at gestational week 33 + 1 by caesarean section. PMID- 19558627 TI - Evaluation of renal artery stenosis in dialysis patients. AB - Atherosclerotic renal artery stenosis (RAS), recognized as a contributor to chronic kidney disease (CKD), may be present in a substantial fraction of dialysis patients. It is generally unknown what proportion of end-stage renal disease patients on dialysis could recover kidney function if RAS were treated. Patients with CKD are often inadequately screened for RAS because of technical limitations of various screening modalities. Multiple small case series and studies have evaluated the role of revascularization for renal salvage in dialysis patients with RAS; these studies are reviewed. Large prospective, randomized multicenter trials of intervention for RAS exclude patients with advanced CKD. The risks of intervention must be weighed against the potential for renal recovery, even when predictors of success are not known with certainty. PMID- 19558628 TI - In good conscience--safely withholding dialysis in the elderly. AB - Across the world, the incidence of end-stage kidney disease is increasing in the elderly. However, they do not always fare very well on renal replacement therapy. Age at the start of dialysis, multiple comorbidities (especially if ischemic heart disease is one of them), diabetes, functional dependence, poor intellectual capacity, low serum albumin, peripheral vascular disease, and late referral have been associated with increased mortality on dialysis in various studies. Moreover, renal failure is only one of the many problems affecting the elderly and dialysis can potentially impair their quality of life tremendously. Therefore, it is often a challenge for the nephrologist to decide whether starting dialysis is in the best interest of the elderly patient. Is it sometimes nobler to provide supportive care without dialysis to an elderly patient with renal failure? Can dialysis be safely delayed where the nephrologist is uncertain of the prognosis or the patient is unsure whether or not to have dialysis? How robust is the evidence base to help inform discussion between the nephrologist and the patient/carer? What are the limitations in carrying out further research in this area? What does conservative management, which is better termed nondialytic supportive care, entail and how should it be delivered? This article aims to answer these fundamental questions confronting the nephrologist in day to day clinical practice. PMID- 19558629 TI - Red blood cell damage from extracorporeal circulation in hemodialysis. AB - Blood damage is an unavoidable side effect of extracorporeal circulation. The effects of blood damage on patients' hematocrit and erythropoietin requirement as well as other potential side effects have not been studied for uneventful treatments. Comparing long nocturnal dialysis with regular 4-hour, three times per week dialysis allows for the conclusion that the influence of blood damage caused by extracorporeal circulation is small compared with biochemical effects. Acute hemolysis is one of the few remaining mechanical problems of dialysis. Acute hemolysis is caused by obstructions within the extracorporeal circuit caused by manufacturing errors, kinking of blood tubing or user errors, or by a combination of excessive flow and improper cannula or catheter dimensions. The risk of acute hemolysis can be further reduced by industrial quality control, better design of dialysis equipment, and hemodialysis machine control. Adverse effects caused by chronic mechanical hemolysis need to be studied. PMID- 19558630 TI - Sphincter repair for incontinence. PMID- 19558634 TI - Necrotizing enterocolitis in neonates receiving octreotide for the management of congenital hyperinsulinism. AB - The somatostatin analog octreotide was used for the first time in the treatment of an infant with congenital hyperinsulinism in 1986. Since then, it is commonly used in the management of congenital hyperinsulinemic hypoglycemias. Despite a wide variety of potential adverse reactions, octreotide is generally well tolerated. It has been extensively demonstrated that octreotide reduces the splanchnic blood flow in a dose-dependent manner, affecting the entire gastrointestinal tract, and some concern has been recently raised regarding the potential implications of this effect in the development of necrotizing enterocolitis in neonates receiving octreotide for the management of congenital hyperinsulinism. The aim of this report is to present a series of patients treated at our institution in which we observed this association, and review the current related literature. PMID- 19558635 TI - MITF pathway mutations in melanoma. PMID- 19558636 TI - Anesthesia and perioperative medical management of children with spinal muscular atrophy. AB - OBJECTIVE: To describe the perioperative medical care, anesthetic considerations, and the risk of postanesthetic respiratory failure in patients with pediatric spinal muscular atrophy (SMA). METHODS: There is a retrospective chart review carried out at an urban, tertiary care pediatric hospital. All patients with ICD 9 codes corresponding to SMA diagnoses were identified, and records screened for anesthetic and perioperative medical management. Medical records were reviewed for demographic, clinical, and outcome data. MAIN RESULTS: Twenty-five children with SMA (10 type I, 8 type II, 7 type III) accounted for 56 general and regional anesthetic cases. Twenty-one (38%) cases had preexisting ventilator dependence. Total intravenous anesthesia with nitrous oxide was provided in 14 (25%) cases, balanced anesthesia with inhalational agents and epidural or systemic opioids were used in 41 (84%) cases, and one infant received a spinal anesthesia. Intraoperative and postoperative complications occurred in 2 (4%) and 2 (4%) cases respectively. One case required an unplanned postanesthetic intensive care unit admission, and there were two late deaths with redirection of care. CONCLUSIONS: Perioperative care can be provided for children with SMA safely and effectively with total intravenous or inhaled anesthetics along with the judicious use of opioids to improve patient comfort without increased morbidity. Further prospective studies of standardized anesthetic and perioperative respiratory medical management in this population are required to minimize the decompensation of tenuous preoperative respiratory function. PMID- 19558637 TI - Recent trends in breast cancer incidence in US white women by county-level urban/rural and poverty status. AB - BACKGROUND: Unprecedented declines in invasive breast cancer rates occurred in the United States between 2001 and 2004, particularly for estrogen receptor positive tumors among non-Hispanic white women over 50 years. To understand the broader public health import of these reductions among previously unstudied populations, we utilized the largest available US cancer registry resource to describe age-adjusted invasive and in situ breast cancer incidence trends for non Hispanic white women aged 50 to 74 years overall and by county-level rural/urban and poverty status. METHODS: We obtained invasive and in situ breast cancer incidence data for the years 1997 to 2004 from 29 population-based cancer registries participating in the North American Association of Central Cancer Registries resource. Annual age-adjusted rates were examined overall and by rural/urban and poverty of patients' counties of residence at diagnosis. Joinpoint regression was used to assess trends by annual quarter of diagnosis. RESULTS: Between 2001 and 2004, overall invasive breast cancer incidence fell 13.2%, with greater reductions among women living in urban (-13.8%) versus rural (-7.5%) and low- (-13.0%) or middle- (-13.8%) versus high- (-9.6%) poverty counties. Most incidence rates peaked around 1999 then declined after second quarter 2002, although in rural counties, rates decreased monotonically after 1999. Similar but more attenuated patterns were seen for in situ cancers. CONCLUSION: Breast cancer rates fell more substantially in urban and low-poverty, affluent counties than in rural or high-poverty counties. These patterns likely reflect a major influence of reductions in hormone therapy use after July 2002 but cannot exclude possible effects due to screening patterns, particularly among rural populations where hormone therapy use was probably less prevalent. PMID- 19558639 TI - Macrophage migration inhibitory factor in cerebrospinal fluid from patients with central nervous system infection. AB - INTRODUCTION: Macrophage migration inhibitory factor (MIF) plays an essential pathophysiological role in septic shock, but its role in central nervous system infection (CNS) remains to be defined. METHODS: We investigated cerebrospinal fluid (CSF) levels of MIF in 171 patients who were clinically suspected of having meningitis on admission. Of these, 31 were found to have purulent meningitis of known aetiology, 20 purulent meningitis of unknown aetiology, 59 lymphocytic meningitis and 11 encephalitis, whereas 50 were suspected of having but had no evidence of CNS infection. RESULTS: CSF MIF levels were significantly higher in patients with purulent meningitis of known aetiology (median [interquartile range]: 8,639 [3,344 to 20,600] ng/l) than in patients with purulent meningitis of unknown aetiology (2,209 [1,516 to 6,550] ng/l; Mann-Whitney test, P = 0.003), patients with lymphocytic meningitis (1,912 [1,302 to 4,105] ng/l; P < 0.001) and patients suspected of having but without evidence of CNS infection (1,472 [672 to 3,447] ng/l; P < 0.001). Also, patients with encephalitis (6,937 [3,961 to 8,353] ng/l) had higher CSF MIF than did patients without CNS infection (P < 0.01). Among patients with purulent meningitis, CSF MIF levels were significantly higher in patients infected with pneumococci than in those with meningococcal infection (11,569 [8,615 to 21,935] ng/l versus 5,006 [1,717 to 10,905] ng/l; P = 0.02), in patients who required versus those not requiring assisted ventilation (10,493 [5,961 to 22,725] ng/l versus 3,240 [1,563 to 9,302] ng/l; P = 0.003), and in patients with versus those without impaired consciousness (8,614 [3,344 to 20,935] ng/l versus 2,625 [1,561 to 7,530] ng/l; P = 0.02). CSF MIF levels correlated significantly with meningeal inflammation (P < 0.05) but not with systemic inflammatory response (P > 0.05) in patients with purulent meningitis of known aetiology, those with lymphocytic meningitis and those with encephalitis. CONCLUSIONS: MIF was significantly increased in the CSF of patients with purulent meningitis and encephalitis, and was to some degree associated with severity of the infection. Our findings indicate that MIF may play an important role in CNS infection. PMID- 19558638 TI - Effect of hydroxyurea on the promoter occupancy profiles of tumor suppressor p53 and p73. AB - BACKGROUND: The p53 tumor suppressor and its related protein, p73, share a homologous DNA binding domain, and mouse genetics studies have suggested that they have overlapping as well as distinct biological functions. Both p53 and p73 are activated by genotoxic stress to regulate an array of cellular responses. Previous studies have suggested that p53 and p73 independently activate the cellular apoptotic program in response to cytotoxic drugs. The goal of this study was to compare the promoter-binding activity of p53 and p73 at steady state and after genotoxic stress induced by hydroxyurea. RESULTS: We employed chromatin immunoprecipitation, the NimbleGen promoter arrays and a model-based algorithm for promoter arrays to identify promoter sequences enriched in anti-p53 or anti p73 immunoprecipitates, either before or after treatment with hydroxyurea, which increased the expression of both p53 and p73 in the human colon cancer cell line HCT116-3(6). We calculated a model-based algorithm for promoter array score for each promoter and found a significant correlation between the promoter occupancy profiles of p53 and p73. We also found that after hydroxyurea treatment, the p53 bound promoters were still bound by p73, but p73 became associated with additional promoters that that did not bind p53. In particular, we showed that hydroxyurea induces the binding of p73 but not p53 to the promoter of MLH3, which encodes a mismatch repair protein, and causes an up-regulation of the MLH3 mRNA. CONCLUSION: These results suggest that hydroxyurea exerts differential effects on the promoter-binding functions of p53 and p73 and illustrate the power of model based algorithm for promoter array in the analyses of promoter occupancy profiles of highly homologous transcription factors. PMID- 19558641 TI - Determinants of the daily rhythm of blood fluidity. AB - BACKGROUND: Numerous processes in the living body exhibit daily rhythmicity. In this study, we characterized a daily rhythm of blood fluidity and identified its determinants. METHODS: The subjects were nine young males. We measured the physiological parameters and performed hematological and biochemical analyses. We repeated the measurements six times during the day at 7:30 (just after getting up and before breakfast), 10:00, 13:30 (after lunch), 16:30, 19:30 (after dinner), and 21:30. The subjects performed sedentary work all day, and the contents and time of the meals were uniform. Investigation of blood rheology was based on Kikuchi's microchannel method. RESULTS: Blood passage time varied significantly with time of day. Stepwise regression analysis was used to determine the significant factors affecting blood passage time. Body temperature, heartbeat, hematocrit, white blood cell and total cholesterol were significant determinants of blood passage time. CONCLUSION: We confirmed that blood fluidity has a daily rhythm. In addition, we found that the determinants of blood fluidity included physiological parameters such as body temperature and heartbeat, hematological parameters such as hematocrit, and white blood cell and total cholesterol. PMID- 19558640 TI - DNA ligase 1 deficient plants display severe growth defects and delayed repair of both DNA single and double strand breaks. AB - BACKGROUND: DNA ligase enzymes catalyse the joining of adjacent polynucleotides and as such play important roles in DNA replication and repair pathways. Eukaryotes possess multiple DNA ligases with distinct roles in DNA metabolism, with clear differences in the functions of DNA ligase orthologues between animals, yeast and plants. DNA ligase 1, present in all eukaryotes, plays critical roles in both DNA repair and replication and is indispensable for cell viability. RESULTS: Knockout mutants of atlig1 are lethal. Therefore, RNAi lines with reduced levels of AtLIG1 were generated to allow the roles and importance of Arabidopsis DNA ligase 1 in DNA metabolism to be elucidated. Viable plants were fertile but displayed a severely stunted and stressed growth phenotype. Cell size was reduced in the silenced lines, whilst flow cytometry analysis revealed an increase of cells in S-phase in atlig1-RNAi lines relative to wild type plants. Comet assay analysis of isolated nuclei showed atlig1-RNAi lines displayed slower repair of single strand breaks (SSBs) and also double strand breaks (DSBs), implicating AtLIG1 in repair of both these lesions. CONCLUSION: Reduced levels of Arabidopsis DNA ligase 1 in the silenced lines are sufficient to support plant development but result in retarded growth and reduced cell size, which may reflect roles for AtLIG1 in both replication and repair. The finding that DNA ligase 1 plays an important role in DSB repair in addition to its known function in SSB repair, demonstrates the existence of a previously uncharacterised novel pathway, independent of the conserved NHEJ. These results indicate that DNA ligase 1 functions in both DNA replication and in repair of both ss and dsDNA strand breaks in higher plants. PMID- 19558642 TI - Characterization of the repertoire diversity of the Plasmodium falciparum stevor multigene family in laboratory and field isolates. AB - BACKGROUND: The evasion of host immune response by the human malaria parasite Plasmodium falciparum has been linked to expression of a range of variable antigens on the infected erythrocyte surface. Several genes are potentially involved in this process with the var, rif and stevor multigene families being the most likely candidates and coding for rapidly evolving proteins. The high sequence diversity of proteins encoded by these gene families may have evolved as an immune evasion strategy that enables the parasite to establish long lasting chronic infections. Previous findings have shown that the hypervariable region (HVR) of STEVOR has significant sequence diversity both within as well as across different P. falciparum lines. However, these studies did not address whether or not there are ancestral stevor that can be found in different parasites. METHODS: DNA and RNA sequences analysis as well as phylogenetic approaches were used to analyse the stevor sequence repertoire and diversity in laboratory lines and Kilifi (Kenya) fresh isolates. RESULTS: Conserved stevor genes were identified in different P. falciparum isolates from different global locations. Consistent with previous studies, the HVR of the stevor gene family was found to be highly divergent both within and between isolates. Importantly phylogenetic analysis shows some clustering of stevor sequences both within a single parasite clone as well as across different parasite isolates. CONCLUSION: This indicates that the ancestral P. falciparum parasite genome already contained multiple stevor genes that have subsequently diversified further within the different P. falciparum populations. It also confirms that STEVOR is under strong selection pressure. PMID- 19558644 TI - Abnormal auditory ERP N100 in children with dyslexia: comparison with their control siblings. AB - BACKGROUND: Recent research has implicated deficits of the working memory (WM) and attention in dyslexia. The N100 component of event-related potentials (ERP) is thought to reflect attention and working memory operation. However, previous studies showed controversial results concerning the N100 in dyslexia. Variability in this issue may be the result of inappropriate match up of the control sample, which is usually based exclusively on age and gender. METHODS: In order to address this question the present study aimed at investigating the auditory N100 component elicited during a WM test in 38 dyslexic children in comparison to those of 19 unaffected sibling controls. Both groups met the criteria of the International Classification of Diseases (ICD-10). ERP were evoked by two stimuli, a low (500 Hz) and a high (3000 Hz) frequency tone indicating forward and reverse digit span respectively. RESULTS: As compared to their sibling controls, dyslexic children exhibited significantly reduced N100 amplitudes induced by both reverse and forward digit span at Fp1, F3, Fp2, Fz, C4, Cz and F4 and at Fp1, F3, C5, C3, Fz, F4, C6, P4 and Fp2 leads respectively. Memory performance of the dyslexics group was not significantly lower than that of the controls. However, enhanced memory performance in the control group is associated with increased N100 amplitude induced by high frequency stimuli at the C5, C3, C6 and P4 leads and increased N100 amplitude induced by low frequency stimuli at the P4 lead. CONCLUSION: The present findings are in support of the notion of weakened capture of auditory attention in dyslexia, allowing for a possible impairment in the dynamics that link attention with short memory, suggested by the anchoring-deficit hypothesis. PMID- 19558643 TI - Which urine marker test provides more diagnostic value in conjunction with standard cytology- ImmunoCyt/uCyt+ or Cytokeratin 20 expression. AB - BACKGROUND: Because of the poor sensitivity of urinary cytological findings for the diagnosis of especially low grade urinary bladder carcinoma, new molecular diagnostic methods have been proposed. We decided to verify the ImmunoCyt/uCyt+ (UCyt+) test and cytology combination and cytokeratin 20 (CK20) and cytology combination in urine as possible diagnostic and monitoring tool for bladder cancer. METHODS: Evaluation of CK20 expression and UCyt+ was performed in urine of 90 patients of which 54 with bladder cancer with primary/recurrent diagnosis (low grade urothelial carcinoma (LGUC) = 23/8 patients, high grade urothelial carcinoma (HGUC) = 18/5 patients), and 36 patients as control; except of neoplastic bladder disease patients. For the evaluation of the three tests, CK20 and UCyt+ tests were combined with urine cytology and compared with each other. RESULTS: The overall sensitivity detected for each tumor marker was as follows: for urine cytology was 75.9% and UCyt+ was 83.3%, for CK20 70.4%, while the specificity was 66.7% for urine cytology and 86.1% for UCyt+ and 83.3% for CK20. The sensitivity of cytology and UCyt+ combination was higher (88.9%) than the sensitivity cytology and CK20 combination (77.8%). The simultaneous use of the three markers, sensitivity was reaching 92.5%. CONCLUSION: The UCyt+ test and CK20 expression are valid tools for the performance of adjunctive analyses with conventional cytologic examination. PMID- 19558645 TI - Soy consumption and risk of COPD and respiratory symptoms: a case-control study in Japan. AB - BACKGROUND: To investigate the relationship between soy consumption, COPD risk and the prevalence of respiratory symptoms, a case-control study was conducted in Japan. METHODS: A total of 278 eligible patients (244 men and 34 women), aged 50 75 years with COPD diagnosed within the past four years, were referred by respiratory physicians, while 340 controls (272 men and 68 women) were recruited from the community. All participants underwent spirometric measurements of respiratory function. Information on demographics, lifestyle characteristics and habitual food consumption was obtained using a structured questionnaire. RESULTS: Total soy consumption was positively correlated with observed lung function measures. The mean soy intake was significantly higher among controls (59.98, SD 50.23 g/day) than cases (44.84, SD 28.5 g/day). A significant reduction in COPD risk was evident for highest versus lowest quartile of daily intake of total soybean products, with adjusted odds ratio (OR) 0.392, 95% CI 0.194-0.793, p for trend 0.001. Similar decreases in COPD risk were associated with frequent and higher intake of soy foods such as tofu and bean sprouts, whereas respiratory symptoms were inversely associated with high consumption of soy foods, especially for breathlessness (OR 0.989, 95% CI 0.982-0.996). CONCLUSION: Increasing soy consumption was associated with a decreased risk of COPD and breathlessness. PMID- 19558646 TI - Divergent fifteen-year trends in traditional and cardiometabolic risk factors of cardiovascular diseases in the Seychelles. AB - OBJECTIVE: Few studies have assessed secular changes in the levels of cardiovascular risk factors (CV-RF) in populations of low or middle income countries. The systematic collection of a broad set of both traditional and metabolic CV-RF in 1989 and 2004 in the population of the Seychelles islands provides a unique opportunity to examine trends at a fairly early stage of the "diabesity" era in a country in the African region. METHODS: Two examination surveys were conducted in independent random samples of the population aged 25-64 years in 1989 and 2004, attended by respectively 1081 and 1255 participants (participation rates >80%). All results are age-standardized to the WHO standard population. RESULTS: In 2004 vs. 1989, the levels of the main traditional CV-RF have either decreased, e.g. smoking (17% vs. 30%, p < 0.001), mean blood pressure (127.8/84.8 vs. 130.0/83.4 mmHg, p < 0.05), or only moderately increased, e.g. median LDL-cholesterol (3.58 vs. 3.36 mmol/l, p < 0. 01). In contrast, marked detrimental trends were found for obesity (37% vs. 21%, p < 0.001) and several cardiometabolic CVD-RF, e.g. mean HDL-cholesterol (1.36 vs. 1.40 mmol/l, p < 0.05), median triglycerides (0.80 vs. 0.78 mmol/l, p < 0.01), mean blood glucose (5.89 vs. 5.22 mmol/l, p < 0.001), median insulin (11.6 vs. 8.3 micromol/l, p < 0.001), median HOMA-IR (2.9 vs. 1.8, p < 0.001) and diabetes (9.4% vs. 6.2%, p < 0.001). At age 40-64, the prevalence of elevated total cardiovascular risk tended to decrease (e.g. WHO-ISH risk score > or =10; 11% vs. 13%, ns), whereas the prevalence of the metabolic syndrome (which integrates several cardiometabolic CVD-RF) nearly doubled (36% vs. 20%, p < 0.001). Data on physical activity and on intake of alcohol, fruit and vegetables are also provided. Awareness and treatment rates improved substantially for hypertension and diabetes, but control rates improved for the former only. Median levels of the cardiometabolic CVD-RF increased between 1989 and 2004 within all BMI strata, suggesting that the worsening levels of cardiometabolic CVD-RF in the population were not only related to increasing BMI levels in the interval. CONCLUSION: The levels of several traditional CVD-RF improved over time, while marked detrimental trends were observed for obesity, diabetes and several cardiometabolic factors. Thus, in this population, the rapid health transition was characterized by substantial changes in the patterns of CVD-RF. More generally, this analysis suggests the importance of surveillance systems to identify risk factor trends and the need for preventive strategies to promote healthy lifestyles and nutrition. PMID- 19558647 TI - Phylo-evo-devo: combining phylogenetics with evolutionary developmental biology. AB - As a result of the integration of molecular and morphological approaches for the reconstruction of phylogenies, and of the intertwining of developmental and evolutionary biology, further prospects are open for a fruitful interaction between these two fields in what we may call a phylo-evo-devo approach.Wiegmann et al.'s molecular phylogeny of the holometabolous insect orders, recently published in BMC Biology, offers a good opportunity to revisit the inverted positions of wings and halteres in the Diptera and the Strepsiptera in terms of a putative homeotic mutation in the Hox gene Ultrabithorax. The main finding of this paper is that Strepsiptera are closely related to the Coleoptera rather than Diptera, as recently claimed. Through this exemplary case, the paper demonstrates the value of the reciprocal illumination we can expect from the integration of a good phylogeny and a sound knowledge of the evolvability of developmental mechanisms. PMID- 19558648 TI - Infectious salmon anaemia virus (ISAV) isolated from the ISA disease outbreaks in Chile diverged from ISAV isolates from Norway around 1996 and was disseminated around 2005, based on surface glycoprotein gene sequences. AB - BACKGROUND: Infectious salmon anaemia (ISA) virus (ISAV) is a pathogen of marine farmed Atlantic salmon (Salmo salar); a disease first diagnosed in Norway in 1984. For over 25 years ISAV has caused major disease outbreaks in the Northern hemisphere, and remains an emerging fish pathogen because of the asymptomatic infections in marine wild fish and the potential for emergence of new epidemic strains. ISAV belongs to the family Orthomyxoviridae, together with influenza viruses but is sufficiently different to be assigned to its own genus, Isavirus. The Isavirus genome consists of eight single-stranded RNA species, and the virions have two surface glycoproteins; fusion (F) protein encoded on segment 5 and haemagglutinin-esterase (HE) protein encoded on segment 6. However, comparison between different ISAV isolates is complicated because there is presently no universally accepted nomenclature system for designation of genetic relatedness between ISAV isolates. The first outbreak of ISA in marine-farmed Atlantic salmon in the Southern hemisphere occurred in Chile starting in June 2007. In order to describe the molecular characteristics of the virus so as to understand its origins, how ISAV isolates are maintained and spread, and their virulence characteristics, we conducted a study where the viral sequences were directly amplified, cloned and sequenced from tissue samples collected from several ISA-affected fish on the different fish farms with confirmed or suspected ISA outbreaks in Chile. This paper describes the genetic characterization of a large number of ISAV strains associated with extensive outbreaks in Chile starting in June 2007, and their phylogenetic relationships with selected European and North American isolates that are representative of the genetic diversity of ISAV. RESULTS: RT-PCR for ISAV F and HE glycoprotein genes was performed directly on tissue samples collected from ISA-affected fish on different farms among 14 fish companies in Chile during the ISA outbreaks that started in June 2007. The genes of the F and HE glycoproteins were cloned and sequenced for 51 and 78 new isolates, respectively. An extensive comparative analysis of ISAV F and HE sequence data, including reference isolates sampled from Norway, Faroe Islands, Scotland, USA, and Canada was performed. Based on phylogenetic analysis of concatenated ISAV F and HE genes of 103 individual isolates, the isolates from the ISA outbreaks in Chile grouped in their own cluster of 7 distinct strains within Genotype I (European genotype) of ISAV, with the closest relatedness to Norwegian ISAVs isolated in 1997. The phylogenetic software program, BACKTRACK, estimated the Chile isolates diverged from Norway isolates about 1996 and, therefore, had been present in Chile for some time before the recent outbreaks. Analysis of the deduced F protein sequence showed 43 of 51 Chile isolates with an 11-amino acid insert between 265N and 266Q, with 100% sequence identity with Genotype I ISAV RNA segment 2. Twenty four different HE-HPRs, including HPR0, were detected, with HPR7b making up 79.7%. This is considered a manifestation of ISAV quasispecies HE protein sequence diversity. CONCLUSION: Taken together, these findings suggest that the ISA outbreaks were caused by virus that was already present in Chile that mutated to new strains. This is the first comprehensive report tracing ISAV from Europe to South America. PMID- 19558650 TI - Are immigrant populations aware about their oral health status? A study among immigrants from Ethiopia. AB - BACKGROUND: Evidence from Western countries indicates that there are fundamental discrepancies between self-perceived illness of immigrants and the provision of health care, according to the Western bio-medical health service model. These need to be understood in the planning and implementation stages of public health care programs for new immigrants. The objectives of the present study were to investigate self-perceived versus clinically diagnosed dental and periodontal health status among immigrants from Ethiopia. METHODS: During 2004-2005, dental and periodontal health status was recorded among 340 Ethiopian immigrants, utilizing the DMFT and CPI indices. Additionally, participants were interviewed using a questionnaire which included perceived dental and periodontal health status. Sensitivity and specificity levels of this perception were calculated and compared with the published scientific literature. RESULTS: Regarding dental caries, according to the three operational cut-off points, sensitivity ranged from 70% to 81%, and specificity ranged from 56% to 67%. Regarding periodontal status, 75% of the subjects clinically diagnosed with periodontal pockets self perceived a "bad" health status of gums (sensitivity) and 54% of the subjects diagnosed without periodontal pockets, reported a "good" health status of gums (specificity). These indications of perception levels were higher than a previous study conducted among native born Israelis. CONCLUSION: Minority ethnic groups should not be prejudicially regarded as less knowledgeable. This is illustrated by the unexpected high level of oral health status perception in the present population. Oral health promotion initiatives among immigrants should be based upon optimal descriptive data in order to accomplish the inherent social commitment to these diverse populations. PMID- 19558649 TI - Clustered microRNAs' coordination in regulating protein-protein interaction network. AB - BACKGROUND: MicroRNAs (miRNAs), a growing class of small RNAs with crucial regulatory roles at the post-transcriptional level, are usually found to be clustered on chromosomes. However, with the exception of a few individual cases, so far little is known about the functional consequence of this conserved clustering of miRNA loci. In animal genomes such clusters often contain non homologous miRNA genes. One hypothesis to explain this heterogeneity suggests that clustered miRNAs are functionally related by virtue of co-targeting downstream pathways. RESULTS: Integrating of miRNA cluster information with protein protein interaction (PPI) network data, our research supports the hypothesis of the functional coordination of clustered miRNAs and links it to the topological features of miRNAs' targets in PPI network. Specifically, our results demonstrate that clustered miRNAs jointly regulate proteins in close proximity of the PPI network. The possibility that two proteins yield to this coordinated regulation is negatively correlated with their distance in PPI network. Guided by the knowledge of this preference, we found several network communities enriched with target genes of miRNA clusters. In addition, our results demonstrate that the variance of this propensity can also partly be explained by protein's connectivity and miRNA's conservation. CONCLUSION: In summary, this work supports the hypothesis of intra-cluster coordination and investigates the extent of this coordination. PMID- 19558651 TI - Ab initio and homology based prediction of protein domains by recursive neural networks. AB - BACKGROUND: Proteins, especially larger ones, are often composed of individual evolutionary units, domains, which have their own function and structural fold. Predicting domains is an important intermediate step in protein analyses, including the prediction of protein structures. RESULTS: We describe novel systems for the prediction of protein domain boundaries powered by Recursive Neural Networks. The systems rely on a combination of primary sequence and evolutionary information, predictions of structural features such as secondary structure, solvent accessibility and residue contact maps, and structural templates, both annotated for domains (from the SCOP dataset) and unannotated (from the PDB). We gauge the contribution of contact maps, and PDB and SCOP templates independently and for different ranges of template quality. We find that accurately predicted contact maps are informative for the prediction of domain boundaries, while the same is not true for contact maps predicted ab initio. We also find that gap information from PDB templates is informative, but, not surprisingly, less than SCOP annotations. We test both systems trained on templates of all qualities, and systems trained only on templates of marginal similarity to the query (less than 25% sequence identity). While the first batch of systems produces near perfect predictions in the presence of fair to good templates, the second batch outperforms or match ab initio predictors down to essentially any level of template quality.We test all systems in 5-fold cross validation on a large non-redundant set of multi-domain and single domain proteins. The final predictors are state-of-the-art, with a template-less prediction boundary recall of 50.8% (precision 38.7%) within +/- 20 residues and a single domain recall of 80.3% (precision 78.1%). The SCOP-based predictors achieve a boundary recall of 74% (precision 77.1%) again within +/- 20 residues, and classify single domain proteins as such in over 85% of cases, when we allow a mix of bad and good quality templates. If we only allow marginal templates (max 25% sequence identity to the query) the scores remain high, with boundary recall and precision of 59% and 66.3%, and 80% of all single domain proteins predicted correctly. CONCLUSION: The systems presented here may prove useful in large-scale annotation of protein domains in proteins of unknown structure. The methods are available as public web servers at the address: http://distill.ucd.ie/shandy/ and we plan on running them on a multi-genomic scale and make the results public in the near future. PMID- 19558653 TI - Bifid T waves in leads V2 and V3 in children: a normal variant. AB - INTRODUCTION: The T wave is rarely bifid, apart from patients with long QT syndrome or subjects treated with antiarrhythmic drugs. At times, a U wave partially superimposed upon the T wave is responsible for an apparently bifid T wave. Bifid T waves, in contrast, have been described in normal children in the past, but the phenomenon has not received any attention in recent years, to the extent that it is not mentioned in current textbooks of paediatric cardiology. Aim of the present study was to determine the incidence and clinical counterpart of bifid T waves in a paediatric population. METHODS: We selected 604 consecutive children free from clinically detectable heart disease; subjects whose electrocardiogram showed a bifid T wave underwent a complete clinical and echocardiographic examination. In addition, the electrocardiograms of 110 consecutive adults have also been analyzed. A T wave was considered as bifid whenever it was notched, being the 2 peaks separated from each other by a notch with duration >/= 0.02 sec and voltage >/= 0.05 mV. Moreover, in 7 children with bifid T wave in lead V2 further precordial recordings were obtained: a small electrode was gradually moved from V1 to V3, and 4 additional leads were recorded: 2 between V1 an V2, and 2 between V2 and V3. RESULTS: A bifid T wave was observed in 110 children (18,3%), with a relatively age-related incidence; the highest rate of bifid T waves (53%) occurred in the group of 5-year-old children. The bifid T wave was detected only in lead V2 in 51 cases (46,4%), only in lead V3 in 5 cases (4,6%), in both leads V2 and V3 in 50 cases (45,4%), and in leads other than V2 and V3 in 4 cases (3,6%). In the adult group, none of the examined electrocardiograms showed bifid T waves in any lead.In the bifid T wave paediatric population, the echocardiogram did not reveal any abnormality, apart from 3 subjects which had an asymptomatic mitral valve prolapse; a trivial mitral and/or tricuspid regurgitation detected by color Doppler, as well as a patent foramen ovale in infants, were not considered as abnormal findings. The QTc interval was normal in all of the subjects; the average QTc interval was not different in the bifid T wave population (402 +/- 46 msec) with respect to the control group (407 +/- 39 msec). CONCLUSION: The incidence of bifid T waves in leads V2 and V3 in normal children is high, and awareness of this phenomenon avoids possible misinterpretations leading to a diagnosis of ECG abnormalities. PMID- 19558652 TI - Adjustment Disorder: epidemiology, diagnosis and treatment. AB - BACKGROUND: Adjustment Disorder is a condition strongly tied to acute and chronic stress. Despite clinical suggestion of a large prevalence in the general population and the high frequency of its diagnosis in the clinical settings, there has been relatively little research reported and, consequently, very few hints about its treatments. METHODS: the authors gathered old and current information on the epidemiology, clinical features, comorbidity, treatment and outcome of adjustment disorder by a systematic review of essays published on PUBMED. RESULTS: After a first glance at its historical definition and its definition in the DSM and ICD systems, the problem of distinguishing AD from other mood and anxiety disorders, the difficulty in the definition of stress and the implied concept of 'vulnerability' are considered. Comorbidity of AD with other conditions, and outcome of AD are then analyzed. This review also highlights recent data about trends in the use of antidepressant drugs, evidence on their efficacy and the use of psychotherapies. CONCLUSION: AD is a very common diagnosis in clinical practice, but we still lack data about its rightful clinical entity. This may be caused by a difficulty in facing, with a purely descriptive methods, a "pathogenic label", based on a stressful event, for which a subjective impact has to be considered. We lack efficacy surveys concerning treatment. The use of psychotropic drugs such as antidepressants, in AD with anxious or depressed mood is not properly supported and should be avoided, while the usefulness of psychotherapies is more solidly supported by clinical evidence. To better determine the correct course of therapy, randomized-controlled trials, even for the combined use of drugs and psychotherapies, are needed vitally, especially for the resistant forms of AD. PMID- 19558654 TI - Efficacy of sulphadoxine-pyrimethamine with or without artesunate for the treatment of uncomplicated Plasmodium falciparum malaria in southern Mozambique: a randomized controlled trial. AB - BACKGROUND: An artemisinin-based combination therapy, artesunate (AS) plus sulphadoxine-pyrimethamine (SP), was compared to SP monotherapy to provide evidence of further treatment options in southern Mozambique. METHODS: Between 2003 and 2005, 411 patients over one year and 10 kg with uncomplicated Plasmodium falciparum malaria were randomly allocated SP (25/1.25 mg per kg day 0) or AS/SP (as above plus 4 mg/kg artesunate days 0, 1 and 2). Allocation was concealed, but treatment was open-label except to microscopists. The primary objective was the relative risk of treatment failure, which was assessed using World Health Organization response definitions modified to a 42-day follow-up. RESULTS: Of the 411 subjects enrolled, 359 (87.3%) completed the follow up period (SP n = 175, AS/SP n = 184). A survival analysis including 408 subjects showed that the polymerase chain reaction-adjusted cure rates were 90.4% (95% confidence interval [CI] 84.9%-93.9%) and 98.0% (95% CI 94.8%-99.3%) for SP and AS/SP respectively. Multivariable analysis showed that treatment with AS/SP decreased the relative hazard of treatment failure by 80% compared to SP (hazard ratio [HR] 0.2; 95% CI 0.1-0.6) and age over seven years decreased the relative hazard of failure by 70% (HR 0.3; 95% CI 0.1-0.9), when compared to younger age. However, having a quintuple dhfr/dhps mutation increased the relative hazard of failure compared to fewer mutations (HR 3.2; 95% CI 1.3-7.5) and baseline axillary temperature increased the relative hazard of failure by 50% for each degree C increase (HR 1.5; 95% CI 1.1-2.2). CONCLUSION: While both treatments were efficacious, AS plus SP significantly decreased the relative hazard of treatment failure compared to SP monotherapy Artesunate plus sulphadoxine-pyrimethamine, but not sulphadoxine pyrimethamine monotherapy, met the current WHO criteria of >95% efficacy for policy implementation. TRIAL REGISTRATION: NCT00203736 and NCT00203814. PMID- 19558655 TI - Allergic sensitisation in tuberculosis patients at the time of diagnosis and following chemotherapy. AB - BACKGROUND: It is still a matter of debate whether there is an association between infection with Mycobacterium tuberculosis (M. tuberculosis) and allergy. Previously, we have shown higher levels of specific IgE to different inhalant allergens and total IgE in tuberculosis (TB) patients compared to controls. The objectives of this study were to evaluate a possible change in allergic sensitisation after successful TB treatment and to confirm the finding of our previous study of enhanced allergic sensitisation in TB patients compared to controls in a more controlled setting. Additionally, we wanted to determine the cytokine profile in the same groups and finally to evaluate the association between the presence of Bacillus Calmette-Guerin vaccination (BCG) scar and allergic sensitisation among the controls. METHODS: Sera were analysed for specific IgE to inhalant allergens (Phadiatop) and total IgE by the use of ImmunoCAP 1000 (Pharmacia Diagnostics). Thirteen different cytokines were also analysed in the sera by multiplex bead immunoassay (Luminex 100, Luminex Corporation), and clinical symptoms of allergy and BCG scar were reported in a questionnaire. RESULTS: A reduction in levels of specific and total IgE were observed after successful TB treatment. TB patients also had higher levels of specific and total IgE compared to healthy controls. Both interleukin (IL)-6 and interferon (IFN)gamma were higher in TB patients compared to healthy controls. The levels of IL-6 were reduced after successful TB treatment. The presence of a BCG scar was associated with a reduced risk of developing allergic sensitisation. CONCLUSION: We observed a reduced level of allergic sensitisation after successful TB treatment. TB patients seem to be more allergically sensitised than healthy controls, confirming our previous finding. Furthermore, we observed an inverse association between allergic sensitisation and visible BCG scar, which adds additional support to the hygiene hypothesis. PMID- 19558656 TI - Respiratory viral infections detected by multiplex PCR among pediatric patients with lower respiratory tract infections seen at an urban hospital in Delhi from 2005 to 2007. AB - BACKGROUND: Acute lower respiratory tract infections (ALRI) are the major cause of morbidity and mortality in young children worldwide. Information on viral etiology in ALRI from India is limited. The aim of the present study was to develop a simple, sensitive, specific and cost effective multiplex PCR (mPCR) assay without post PCR hybridization or nested PCR steps for the detection of respiratory syncytial virus (RSV), influenza viruses, parainfluenza viruses (PIV1 3) and human metapneumovirus (hMPV). Nasopharyngeal aspirates (NPAs) were collected from children with ALRI < or = 5 years of age. The sensitivity and specificity of mPCR was compared to virus isolation by centrifugation enhanced culture (CEC) followed by indirect immunofluorescence (IIF). RESULTS: From April 2005-March 2007, 301 NPAs were collected from children attending the outpatient department or admitted to the ward of All India Institute of Medical Sciences hospital at New Delhi, India. Multiplex PCR detected respiratory viruses in 106 (35.2%) of 301 samples with 130 viruses of which RSV was detected in 61, PIV3 in 22, PIV2 in 17, hMPV in 11, PIV1 in 10 and influenza A in 9 children. CEC-IIF detected 79 viruses only. The sensitivity of mPCR was 0.1TCID50 for RSV and influenza A and 1TCID50 for hMPV, PIV1, PIV2, PIV3 and Influenza B. Mixed infections were detected in 18.8% of the children with viral infections, none detected by CEC-IIF. Bronchiolitis was significantly associated with both total viral infections and RSV infection (p < 0.05). History of ARI in family predisposed children to acquire viral infection (p > 0.05). CONCLUSION: Multiplex PCR offers a rapid, sensitive and reasonably priced diagnostic method for common respiratory viruses. PMID- 19558657 TI - The dihydropyridine calcium channel blocker benidipine prevents lysophosphatidylcholine-induced endothelial dysfunction in rat aorta. AB - BACKGROUND: Lysophosphatidylcholine (LPC), an atherogenic component of oxidized low-density lipoprotein, has been shown to induce the attenuation of endothelium dependent vascular relaxation. Although benidipine, a dihydropyridine-calcium channel blocker, is known to have endothelial protective effects, the effects of benidipine on LPC-induced endothelial dysfunction remain unknown. We examined the effects of benidipine on the impairment of endothelium-dependent relaxation induced by LPC. METHODS: Benidipine was administered orally to rats and aortas were then isolated. Aortic rings were treated with LPC and endothelial functions were then evaluated. Additionally, the effects of benidipine on intracellular calcium concentration ([Ca2+]i) and membrane fluidity altered by LPC in primary cultured rat aortic endothelial cells were examined. [Ca2+]i was measured using the fluorescent calcium indicator fura-2. Membrane fluidity was monitored by measuring fluorescence recovery after photobleaching. RESULTS: Treatment with LPC impaired endothelial function. Benidipine prevents the impairment of relaxation induced by LPC. Acetylcholine elicited an increase in [Ca2+]i in fura-2 loaded endothelial cells. The increase in [Ca2+]i was suppressed after exposure to LPC. Plasma membrane fluidity increased following incubation with LPC. Benidipine inhibited the LPC-induced increase in membrane fluidity and impairment of increase in [Ca2+]i. CONCLUSION: These results suggest that benidipine inhibited LPC-induced endothelial dysfunction by maintaining increase in [Ca2+]i. Benidipine possesses membrane stabilization properties in LPC-treated endothelial cells. It is speculated that the preservation of membrane fluidity by benidipine may play a role in the retainment of calcium mobilization. The present findings may provide new insights into the endothelial protective effects of benidipine. PMID- 19558658 TI - Smoking-attributable mortality and years of potential life lost in 16 Brazilian capitals, 2003: a prevalence-based study. AB - BACKGROUND: To establish the impact of tobacco smoking on mortality is essential to define and monitor public health interventions in developing countries. METHODS: The Smoking-Attributable Mortality, Morbidity and Economic Costs (SAMMEC) software was used to estimate the smoking attributable mortality (SAM) in 15 Brazilian State Capitals and the Federal District for the year 2003. Smoking prevalence and mortality data of people aged 35 years or older were obtained for each city from the Brazilian Household Survey on Non Communicable Diseases Risk Factors (2002-2003) and from the Brazilian Mortality System (2003), respectively. RESULTS: In 2003, of the 177,543 deaths of persons aged 35 years and older 24,222 (13.64%) were attributable to cigarette smoking. This total represents 18.08% of all male deaths (n = 16,896) and 8.71% (n = 7,326) of all female deaths in these cities. The four leading causes of smoking-attributable death were chronic airways obstruction (4,419 deaths), ischemic heart disease (4,417 deaths), lung cancer (3,682 deaths), and cerebrovascular disease (3,202 deaths). Cigarette smoking accounted for 419,935 years of potential life lost (YPLL) (279,990 YPLL for men and 139,945 YPLL for women) in the same period. CONCLUSION: Tobacco use caused one out of five male deaths and one out of ten female deaths in the sixteen cities in 2003. Four leading causes of smoking attributable deaths (ischemic heart disease, chronic airways obstruction, lung cancer and cerebrovascular disease) accounted for 64.9% of SAM. Effective and comprehensive actions must be taken in order to slow this epidemic in Brazil. PMID- 19558659 TI - n-3 polyunsaturated fatty acids in milk is associate to weight gain and growth in premature infants. AB - BACKGROUND: Linoleic 18:2 (n-6) and alpha-linolenic 18:3 (n-3) essential fatty acids and long-chain polyunsaturated fatty acids (LC-PUFA) are essential nutrients for growth and neonatal development. Consumption of preformed n-3 LC PUFA has been shown to increase gestational duration and to decrease the incidence of premature birth in human studies. This study evaluated the association of essential fatty acids and LC-PUFA in breast milk on the growth of premature children (weight, height and head circumference). STUDY DESIGN: Thirty seven premature infants with a gestational age of 37 weeks or less were followed until 6 months of gestational age, adjusted for prematurity. The milk from mothers, weight, height and head circumference measures of children were collected during the follow up. The breast milk fatty acids were quantified by gas-liquid chromatography. RESULTS: Our results showed that total n-3 PUFA was positively associated with weight gain (p = 0.05), height (p = 0.04) and body mass index (BMI) of children (p = 0.05). Our results also indicate that both linoleic acid and total essential fatty acids were positively associated with BMI and head circumference, whereas oleic acid was positively associated only with head circumference. CONCLUSION: These results suggest that the n-3 PUFA composition of milk may be associated with weight gain and growth. Considering the advantages of n-3 LC-PUFA consumption on infant growth and visual function and its association with reduced incidence of premature birth, dietitians should advise pregnant women to increase their intake of foods high in n-3 LC-PUFA. PMID- 19558661 TI - Gluten encephalopathy with psychiatric onset: case report. AB - Many cases of coeliac disease, a gastrointestinal autoimmune disorder caused by sensitivity to gluten, can remain in a subclinical stage or undiagnosed. In a significant proportion of cases (10-15%) gluten intolerance can be associated with central or peripheral nervous system and psychiatric disorders.A 38-year-old man was admitted as to our department an inpatient for worsening anxiety symptoms and behavioural alterations. After the addition of second generation antipsychotic to the therapeutic regimen, the patient presented neuromotor impairment with high fever, sopor, leukocytosis, raised rhabdomyolysis-related indicators. Neuroleptic malignant syndrome was strongly suspected. After worsening of his neuropsychiatric conditions, with the onset of a frontal cognitive deficit, bradykinesia and difficulty walking, dysphagia, anorexia and hypoferraemic anaemia, SPET revealed a reduction of cerebral perfusion and ENeG results were compatible with a mainly motor polyneuropathy. Extensive laboratory investigations gave positive results for anti-gliadin antibodies, and an appropriate diet led to a progressive remission of the encephalopathy. PMID- 19558660 TI - Sex-associated effect of CETP and LPL polymorphisms on postprandial lipids in familial hypercholesterolaemia. AB - BACKGROUND: This study assessed the gender-specific influence of the cholesteryl ester transfer protein (TaqIB, I405V) and lipoprotein lipase (S447X) polymorphisms on the response to an oral fat tolerance test in heterozygotes for familial hypercholesterolaemia. METHODS: We selected and genotyped 80 men and postmenopausal women heterozygous for familial hypercholesterolaemia (main group) as well as 11 healthy control subjects. Patients were subgrouped based on their response to oral fat tolerance test. The oral fat tolerance test was defined as pathological when postprandial triglyceride concentration was higher than the highest triglyceride concentration observed in healthy subjects (220 mg/dl) at any time (2, 4, 6 or 8 h). RESULTS: In the pathological subgroup, men had significantly higher incremental area under the curve after oral fat tolerance test than postmenopausal women. Furthermore, multivariate analysis revealed a gender association of TaqIB and I405V influence on postprandial lipaemia in this subgroup. CONCLUSION: In conclusion, it seems that gender and TaqIB polymorphism of the cholesteryl ester transfer protein gene were both associated with the distribution of triglyceride values after oral fat tolerance test, only in subjects with a pathological response to oral fat tolerance test. Specifically, men carrying the B2 allele of the TaqIB polymorphism showed a higher postprandial triglyceride peak and a delayed return to basal values compared with women carrying B2. However, further investigations in larger populations are required to replicate and confirm these findings. PMID- 19558662 TI - Construction and sequence sampling of deep-coverage, large-insert BAC libraries for three model lepidopteran species. AB - BACKGROUND: Manduca sexta, Heliothis virescens, and Heliconius erato represent three widely-used insect model species for genomic and fundamental studies in Lepidoptera. Large-insert BAC libraries of these insects are critical resources for many molecular studies, including physical mapping and genome sequencing, but not available to date. RESULTS: We report the construction and characterization of six large-insert BAC libraries for the three species and sampling sequence analysis of the genomes. The six BAC libraries were constructed with two restriction enzymes, two libraries for each species, and each has an average clone insert size ranging from 152-175 kb. We estimated that the genome coverage of each library ranged from 6-9 x, with the two combined libraries of each species being equivalent to 13.0-16.3 x haploid genomes. The genome coverage, quality and utility of the libraries were further confirmed by library screening using 6 approximately 8 putative single-copy probes. To provide a first glimpse into these genomes, we sequenced and analyzed the BAC ends of approximately 200 clones randomly selected from the libraries of each species. The data revealed that the genomes are AT-rich, contain relatively small fractions of repeat elements with a majority belonging to the category of low complexity repeats, and are more abundant in retro-elements than DNA transposons. Among the species, the H. erato genome is somewhat more abundant in repeat elements and simple repeats than those of M. sexta and H. virescens. The BLAST analysis of the BAC end sequences suggested that the evolution of the three genomes is widely varied, with the genome of H. virescens being the most conserved as a typical lepidopteran, whereas both genomes of H. erato and M. sexta appear to have evolved significantly, resulting in a higher level of species- or evolutionary lineage-specific sequences. CONCLUSION: The high-quality and large-insert BAC libraries of the insects, together with the identified BACs containing genes of interest, provide valuable information, resources and tools for comprehensive understanding and studies of the insect genomes and for addressing many fundamental questions in Lepidoptera. The sample of the genomic sequences provides the first insight into the constitution and evolution of the insect genomes. PMID- 19558664 TI - Liposarcoma: exploration of clinical prognostic factors for risk based stratification of therapy. AB - BACKGROUND: Prognosis and optimal treatment strategies of liposarcoma have not been fully defined. The purpose of this study is to define the distinctive clinical features of liposarcomas by assessing prognostic factors. METHODS: Between January 1995 and May 2008, 94 liposarcoma patients who underwent surgical resection with curative intent were reviewed. RESULTS: Fifty patients (53.2%) presented with well differentiated, 22 (23.4%) myxoid, 15 (16.0%) dedifferentiated, 5 (5.3%) round cell, and 2 (2.1%) pleomorphic histology. With the median 14 cm sized of tumor burden, about half of the cases were located in the retroperitoneum (46.8%). Seventy two (76.6%) patients remained alive with 78.1%, and 67.5% of the 5- and 10-year overall survival (OS) rates, respectively. Low grade liposarcoma (well differentiated and myxoid) had a significantly prolonged OS and disease free survival (DFS) with adjuvant radiotherapy when compared with those without adjuvant radiotherapy (5-year OS, 100% vs 66.3%, P = 0.03; 1-year DFS, 92.9% vs 50.0%, respectively, P = 0.04). Independent prognostic factors for OS were histologic variant (P = 0.001; HR, 5.1; 95% CI, 2.0 - 12.9), and margin status (P = 0.005; HR, 4.1; 95% CI, 1.6-10.5). We identified three different risk groups: group 1 (n = 66), no adverse factors; group 2, one or two adverse factors (n = 28). The 5-year OS rate for group 1, and 2 were 91.9%, 45.5%, respectively. CONCLUSION: The histologic subtype, and margin status were independently associated with OS, and adjuvant radiotherapy seems to confer survival benefit in low grade tumors. Our prognostic model for primary liposarcoma demonstrated distinct three groups of patients with good prognostic discrimination. PMID- 19558663 TI - 249 TP53 mutation has high prevalence and is correlated with larger and poorly differentiated HCC in Brazilian patients. AB - BACKGROUND: Ser-249 TP53 mutation (249(Ser)) is a molecular evidence for aflatoxin-related carcinogenesis in Hepatocellular Carcinoma (HCC) and it is frequent in some African and Asian regions, but it is unusual in Western countries. HBV has been claimed to add a synergic effect on genesis of this particular mutation with aflatoxin. The aim of this study was to investigate the frequency of 249(Ser) mutation in HCC from patients in Brazil. METHODS: We studied 74 HCC formalin fixed paraffin blocks samples of patients whom underwent surgical resection in Brazil. 249(Ser) mutation was analyzed by RFLP and DNA sequencing. HBV DNA presence was determined by Real-Time PCR. RESULTS: 249(Ser) mutation was found in 21/74 (28%) samples while HBV DNA was detected in 13/74 (16%). 249Ser mutation was detected in 21/74 samples by RFLP assay, of which 14 were confirmed by 249(Ser) mutant-specific PCR, and 12 by nucleic acid sequencing. All HCC cases with p53-249ser mutation displayed also wild-type p53 sequences. Poorly differentiated HCC was more likely to have 249(Ser) mutation (OR = 2.415, 95% CI = 1.001 - 5.824, p = 0.05). The mean size of 249(Ser) HCC tumor was 9.4 cm versus 5.5 cm on wild type HCC (p = 0.012). HBV DNA detection was not related to 249(Ser) mutation. CONCLUSION: Our results indicate that 249(Ser) mutation is a HCC important factor of carcinogenesis in Brazil and it is associated to large and poorly differentiated tumors. PMID- 19558665 TI - Discovery of cis-elements between sorghum and rice using co-expression and evolutionary conservation. AB - BACKGROUND: The spatiotemporal regulation of gene expression largely depends on the presence and absence of cis-regulatory sites in the promoter. In the economically highly important grass family, our knowledge of transcription factor binding sites and transcriptional networks is still very limited. With the completion of the sorghum genome and the available rice genome sequence, comparative promoter analyses now allow genome-scale detection of conserved cis elements. RESULTS: In this study, we identified thousands of phylogenetic footprints conserved between orthologous rice and sorghum upstream regions that are supported by co-expression information derived from three different rice expression data sets. In a complementary approach, cis-motifs were discovered by their highly conserved co-occurrence in syntenic promoter pairs. Sequence conservation and matches to known plant motifs support our findings. Expression similarities of gene pairs positively correlate with the number of motifs that are shared by gene pairs and corroborate the importance of similar promoter architectures for concerted regulation. This strongly suggests that these motifs function in the regulation of transcript levels in rice and, presumably also in sorghum. CONCLUSION: Our work provides the first large-scale collection of cis elements for rice and sorghum and can serve as a paradigm for cis-element analysis through comparative genomics in grasses in general. PMID- 19558666 TI - It's all about recognition! Qualitative study of the value of interpersonal continuity in general practice. AB - BACKGROUND: Continuity of care has traditionally been regarded as a core quality of general practice, but the long-term doctor-patient relationship has been put under pressure. In many places practices are expanding, with larger teams and more registered patients, thereby threatening the possibility of patients staying with their own general practitioner (GP). GPs often take it for granted that interpersonal continuity is valuable. However, little is known about how patient satisfaction is related to interpersonal continuity. The purpose of this study is to explore the creation of patient satisfaction or dissatisfaction in the interpersonal relation with the GP, and in a comprehensive way to investigate how this is related to continuity of care. METHODS: Qualitative study based on 22 interviews with patients from two practices in Denmark. A total of 12 patients saw a regular doctor and 10 saw an unfamiliar doctor. The patients were selected after an observed consultation and sampled purposefully according to reason for encounter, age and sex. Interpretative phenomenological analysis (IPA) was used to study how patients perceive meeting either a regular or an unfamiliar GP. The analysis explored the patients' perception of their interpersonal relationship with their GP, and interpreted the accounts by using social psychological theories. RESULTS: A long-term continuous relationship with the GP could be satisfactory, but it could also be the reverse. The same pattern was shown in case of an unfamiliar GP. Therefore, patient satisfaction and interpersonal continuity were not causally related. On the contrary, there was a general pattern of how the satisfactory and trustful doctor-patient relationship from the patients' point of view could be created, maintained or destroyed. A pattern where the process of recognition, by respecting and remembering, on the one hand created and maintained satisfaction while humiliation on the other hand destroyed satisfaction in the relationship. CONCLUSION: It was not valuable to have a continuous relationship unless the GP recognized the patient. The social psychological concept of recognition had two different meanings and the GP had to do both, respect and remember the patient, in order to create and sustain the trustful relationship. The added value of interpersonal continuity had to be combined with recognition. PMID- 19558667 TI - Rodent-specific alternative exons are more frequent in rapidly evolving genes and in paralogs. AB - BACKGROUND: Alternative splicing is an important mechanism for generating functional and evolutionary diversity of proteins in eukaryotes. Here, we studied the frequency and functionality of recently gained, rodent-specific alternative exons. RESULTS: We projected the data about alternative splicing of mouse genes to the rat, human, and dog genomes, and identified exons conserved in the rat genome, but missing in more distant genomes. We estimated the frequency of rodent specific exons while controlling for possible residual conservation of spurious exons. The frequency of rodent-specific exons is higher among predominantly skipped exons and exons disrupting the reading frame. Separation of all genes by the rate of sequence evolution and by gene families has demonstrated that rodent specific cassette exons are more frequent in rapidly evolving genes and in rodent specific paralogs. CONCLUSION: Thus we demonstrated that recently gained exons tend to occur in fast-evolving genes, and their inclusion rate tends to be lower than that of older exons. This agrees with the theory that gain of alternative exons is one of the major mechanisms of gene evolution. PMID- 19558668 TI - Large sharing networks and unusual injection practices explain the rapid rise in HIV among IDUs in Sargodha, Pakistan. AB - BACKGROUND: Of the nearly 100,000 street-based IDUs in Pakistan, 20% have HIV. We investigated the recent rise in HIV prevalence from 12 to 52% among IDUs in Sargodha despite > 70% coverage with syringe exchanges. METHODS: We interviewed approximately 150 IDUs and 30 outreach workers in focus group discussions. RESULTS: We found six rural and 28 urban injecting locations. Urban locations have about 20-30 people at any time and about 100 daily; rural locations have twice as many (national average: 4-15). About half of the IDUs started injecting within the past 2 years and are not proficient at injecting themselves. They use street injectors, who have 15-16 clients daily. Heroin is almost exclusively the drug used. Most inject 5-7 times daily.Nearly all injectors claim to use fresh syringes. However, they load, inject and share using a locally developed method called scale. Most Pakistani IDUs prefer to double pump drug the syringe, which allows mixing of blood with drug in the syringe. The injector injects 3 ml and keeps 2 ml (the scale) as injection fee. The injector usually pools all the leftover scale (now with some blood mixed with drug) either for his own use or to sell it. Most IDUs backload the scale they buy into their own fresh syringes. DISCUSSION: Use of an unprecedented method of injecting drugs that largely bypasses fresh syringes, larger size of sharing networks, higher injection frequency and near universal use of street injectors likely explain for the rapid rise in HIV prevalence among IDUs in Sargodha despite high level provision of fresh syringes. This had been missed by us and the national surveillance, which is quantitative. We have addressed this by hiring injectors as peer outreach workers and increasing syringe supply. Our findings highlight both the importance of qualitative research and operations research to enrich the quality of HIV prevention programs. PMID- 19558669 TI - Lymphocyte apoptosis in murine Pneumocystis pneumonia. AB - BACKGROUND: Apoptosis of lymphocytes is important in the termination of an immune response to infection but has also been shown to have detrimental effects in animal models of systemic infection and sepsis. We sought to characterize lymphocyte apoptosis in an animal model of pneumonia due to Pneumocystis murina, an infection localized to the lungs. METHODS: Control mice and mice depleted of CD4+ lymphocytes were inoculated with Pneumocystis. Apoptosis of lung and spleen lymphocytes was assayed by flow cytometry and PCR assay of apoptotic proteins. RESULTS: In control mice, apoptosis of lung lymphocytes was maximal just after the infection was cleared from lung tissue and then declined. However, in CD4 depleted mice, apoptosis was also upregulated in recruited lymphocytes in spite of progressive infection. In splenic lymphocytes, apoptosis was observed early at 1 week after inoculation and then declined. Apoptosis of lung lymphocytes in control mice was associated with a decrease in mRNA for Bcl-2 and an increase in mRNA for Bim. In CD4-depleted mice, lavaged CD8+ cells did change intracellular Bcl-2 but showed increased mRNA for Bim. CONCLUSION: Apoptosis of both pulmonary and extrapulmonary lymphocytes is part of the normal host response to Pneumocystis but is also triggered in CD4-deficient animals with progressive infection. In normal mice apoptosis of pulmonary lymphocytes may serve to terminate the immune response in lung tissue. Apoptosis of lung lymphocytes takes place via both the intrinsic and extrinsic apoptotic pathways and is associated with changes in both pro- and anti-apoptotic proteins. PMID- 19558670 TI - Possible mechanisms of kidney repair. AB - In most adult epithelia the process of replacing damaged or dead cells is maintained through the presence of stem/progenitor cells, which allow epithelial tissues to be repaired following injury. Existing evidence strongly supports the presence of stem cells in the adult kidney. Indeed, recent findings provide evidence in favour of a role for intrinsic renal cells and against a physiological role for bone marrow-derived stem cells in the regeneration of renal epithelial cells. In addition, recent studies have identified a subset of CD24+CD133+ renal progenitors within the Bowman's capsule of adult human kidney, which provides regenerative potential for injured renal epithelial cells. Intriguingly, CD24+CD133+ renal progenitors also represent common progenitors of tubular cells and podocytes during renal development. Chronic injury causes dysfunction of the tubular epithelial cells, which triggers the release of fibrogenic cytokines and recruitment of inflammatory cells to injured kidneys. The rapid interposition of scar tissue probably confers a survival advantage by preventing infectious microorganisms from invading the wound, but prevents subsequent tissue regeneration. However, the existence of renal epithelial progenitors in the kidney suggests a possible explanation for the regression of renal lesions which has been observed in experimental animals and even in humans. Thus, manipulation of the wound repair process in order to shift it towards regeneration will probably require the ability to slow the rapid fibrotic response so that renal progenitor cells can allow tissue regeneration rather than scar formation. PMID- 19558672 TI - Assessment of safety and feasibility of a new technical variant of gastropexy for percutaneous endoscopic gastrostomy: an experience with 435 cases. AB - BACKGROUND: Percutaneous Endoscopic Gastrostomy (PEG) performed through the Introducer Technique is associated with lower risk of surgical infection when compared to the Pull Technique. Its use is less widespread as the fixation of the stomach to the abdominal wall is a stage of the procedure that is difficult to be performed. We present a new technical variant of gastropexy which is fast and easy to be performed. The aim of this study was to evaluate the safety and feasibility of a new technical variant of gastropexy in patients submitted to gastrostomy performed through the Introducer Technique. METHODS: All the patients submitted to PEG through the Introducer Technique were evaluated using a new technical variant of gastropexy, which consists of two parallel stitches of trasfixation sutures involving the abdominal wall and the gastric wall, performed with a long curved needle. Prophylactic antibiotics were not used. Demographic aspects, initial diagnosis, indication, sedation doses, morbidity and surgical mortality were all analyzed. RESULTS: Four hundred and thirty-five consecutive PEGs performed between June 2004 and May 2007 were studied. Nearly all the cases consisted of patients presenting malignant neoplasia, 79.5% of which sited in the head and neck. The main indication of PEG was dysphagia, found in 346 patients (79.5%). There were 12 complications (2.8%) in 11 patients, from which only one patient had peristomal infection (0.2%). There was one death related to the procedure. CONCLUSION: Gastropexy with the technical variant described here is easy to be performed and was feasible and safe in the present study. PEG performed by the Introducer Technique with this type of gastropexy was associated with low rates of wound infection even without the use of prophylactic antibiotics. PMID- 19558671 TI - Oleic acid and peanut oil high in oleic acid reverse the inhibitory effect of insulin production of the inflammatory cytokine TNF-alpha both in vitro and in vivo systems. AB - BACKGROUND: Chronic inflammation is a key player in pathogenesis. The inflammatory cytokine, tumor necrosis factor-alpha is a well known inflammatory protein, and has been a therapeutic target for the treatment of diseases such as Rheumatoid Arthritis and Crohn's Disease. Obesity is a well known risk factor for developing non-insulin dependent diabetes melitus. Adipose tissue has been shown to produce tumor necrosis factor-alpha, which has the ability to reduce insulin secretion and induce insulin resistance. Based on these observations, we sought to investigate the impact of unsaturated fatty acids such as oleic acid in the presence of TNF-alpha in terms of insulin production, the molecular mechanisms involved and the in vivo effect of a diet high in oleic acid on a mouse model of type II diabetes, KKAy. METHODS: The rat pancreatic beta cell line INS-1 was used as a cell biological model since it exhibits glucose dependent insulin secretion. Insulin production assessment was carried out using enzyme linked immunosorbent assay and cAMP quantification with competitive ELISA. Viability of TNF-alpha and oleic acid treated cells was evaluated using flow cytometry. PPAR-gamma translocation was assessed using a PPRE based ELISA system. In vivo studies were carried out on adult male KKAy mice and glucose levels were measured with a glucometer. RESULTS: Oleic acid and peanut oil high in oleic acid were able to enhance insulin production in INS-1. TNF-alpha inhibited insulin production but pre-treatment with oleic acid reversed this inhibitory effect. The viability status of INS-1 cells treated with TNF-alpha and oleic acid was not affected. Translocation of the peroxisome proliferator- activated receptor transcription factor to the nucleus was elevated in oleic acid treated cells. Finally, type II diabetic mice that were administered a high oleic acid diet derived from peanut oil, had decreased glucose levels compared to animals administered a high fat diet with no oleic acid. CONCLUSION: Oleic acid was found to be effective in reversing the inhibitory effect in insulin production of the inflammatory cytokine TNF-alpha. This finding is consistent with the reported therapeutic characteristics of other monounsaturated and polyunsaturated fatty acids. Furthermore, a diet high in oleic acid, which can be easily achieved through consumption of peanuts and olive oil, can have a beneficial effect in type II diabetes and ultimately reverse the negative effects of inflammatory cytokines observed in obesity and non insulin dependent diabetes mellitus. PMID- 19558673 TI - Protection from pulmonary ischemia-reperfusion injury by adenosine A2A receptor activation. AB - BACKGROUND: Lung ischemia-reperfusion (IR) injury leads to significant morbidity and mortality which remains a major obstacle after lung transplantation. However, the role of various subset(s) of lung cell populations in the pathogenesis of lung IR injury and the mechanisms of cellular protection remain to be elucidated. In the present study, we investigated the effects of adenosine A2A receptor (A2AAR) activation on resident lung cells after IR injury using an isolated, buffer-perfused murine lung model. METHODS: To assess the protective effects of A2AAR activation, three groups of C57BL/6J mice were studied: a sham group (perfused for 2 hr with no ischemia), an IR group (1 hr ischemia + 1 hr reperfusion) and an IR+ATL313 group where ATL313, a specific A2AAR agonist, was included in the reperfusion buffer after ischemia. Lung injury parameters and pulmonary function studies were also performed after IR injury in A2AAR knockout mice, with or without ATL313 pretreatment. Lung function was assessed using a buffer-perfused isolated lung system. Lung injury was measured by assessing lung edema, vascular permeability, cytokine/chemokine activation and myeloperoxidase levels in the bronchoalveolar fluid. RESULTS: After IR, lungs from C57BL/6J wild type mice displayed significant dysfunction (increased airway resistance, pulmonary artery pressure and decreased pulmonary compliance) and significant injury (increased vascular permeability and edema). Lung injury and dysfunction after IR were significantly attenuated by ATL313 treatment. Significant induction of TNF-alpha, KC (CXCL1), MIP-2 (CXCL2) and RANTES (CCL5) occurred after IR which was also attenuated by ATL313 treatment. Lungs from A2AAR knockout mice also displayed significant dysfunction, injury and cytokine/chemokine production after IR, but ATL313 had no effect in these mice. CONCLUSION: Specific activation of A2AARs provides potent protection against lung IR injury via attenuation of inflammation. This protection occurs in the absence of circulating blood thereby indicating a protective role of A2AAR activation on resident lung cells such as alveolar macrophages. Specific A2AAR activation may be a promising therapeutic target for the prevention or treatment of pulmonary graft dysfunction in transplant patients. PMID- 19558675 TI - Identification of proteins involved in neural progenitor cell targeting of gliomas. AB - BACKGROUND: Glioblastoma are highly aggressive tumors with an average survival time of 12 months with currently available treatment. We have previously shown that specific embryonic neural progenitor cells (NPC) have the potential to target glioma growth in the CNS of rats. The neural progenitor cell treatment can cure approximately 40% of the animals with malignant gliomas with no trace of a tumor burden 6 months after finishing the experiment. Furthermore, the NPCs have been shown to respond to signals from the tumor environment resulting in specific migration towards the tumor. Based on these results we wanted to investigate what factors could influence the growth and progression of gliomas in our rodent model. METHODS: Using microarrays we screened for candidate genes involved in the functional mechanism of tumor inhibition by comparing glioma cell lines to neural progenitor cells with or without anti-tumor activity. The expression of candidate genes was confirmed at RNA level by quantitative RT-PCR and at the protein level by Western blots and immunocytochemistry. Moreover, we have developed in vitro assays to mimic the antitumor effect seen in vivo. RESULTS: We identified several targets involved in glioma growth and migration, specifically CXCL1, CD81, TPT1, Gas6 and AXL proteins. We further showed that follistatin secretion from the NPC has the potential to decrease tumor proliferation. In vitro co-cultures of NPC and tumor cells resulted in the inhibition of tumor growth. The addition of antibodies against proteins selected by gene and protein expression analysis either increased or decreased the proliferation rate of the glioma cell lines in vitro. CONCLUSION: These results suggest that these identified factors might be useful starting points for performing future experiments directed towards a potential therapy against malignant gliomas. PMID- 19558676 TI - Assessing the internal validity of a household survey-based food security measure adapted for use in Iran. AB - BACKGROUND: The prevalence of food insecurity is an indicator of material well being in an area of basic need. The U.S. Food Security Module has been adapted for use in a wide variety of cultural and linguistic settings around the world. We assessed the internal validity of the adapted U.S. Household Food Security Survey Module to measure adult and child food insecurity in Isfahan, Iran, using statistical methods based on the Rasch measurement model. METHODS: The U.S. Household Food Security Survey Module was translated into Farsi and after adaptation, administered to a representative sample. Data were provided by 2,004 randomly selected households from all sectors of the population of Isfahan, Iran, during 2005. RESULTS: 53.1 percent reported that their food had run out at some time during the previous 12 months and they did not have money to buy more, while 26.7 percent reported that an adult had cut the size of a meal or skipped a meal because there was not enough money for food, and 7.2 percent reported that an adult did not eat for a whole day because there was not enough money for food. The severity of the items in the adult scale, estimated under Rasch-model assumptions, covered a range of 6.65 logistic units, and those in the child scale 11.68 logistic units. Most Iteminfit statistics were near unity, and none exceeded 1.20. CONCLUSION: The range of severity of items provides measurement coverage across a wide range of severity of food insecurity for both adults and children. Both scales demonstrated acceptable levels of internal validity, although several items should be improved. The similarity of the response patterns in the Isfahan and the U.S. suggests that food insecurity is experienced, managed, and described similarly in the two countries. PMID- 19558677 TI - Modeling AIDS survival after initiation of antiretroviral treatment by Weibull models with changepoints. AB - BACKGROUND: Mortality of HIV-infected patients initiating antiretroviral therapy in the developing world is very high immediately after the start of ART therapy and drops sharply thereafter. It is necessary to use models of survival time that reflect this change. METHODS: In this endeavor, parametric models with changepoints such as Weibull models can be useful in order to explicitly model the underlying failure process, even in the case where abrupt changes in the mortality rate are present. Estimation of the temporal location of possible mortality changepoints has important implications on the effective management of these patients. We briefly describe these models and apply them to the case of estimating survival among HIV-infected patients who are initiating antiretroviral therapy in a care and treatment programme in sub-Saharan Africa. RESULTS: As a first reported data-driven estimate of the existence and location of early mortality changepoints after antiretroviral therapy initiation, we show that there is an early change in risk of death at three months, followed by an intermediate risk period lasting up to 10 months after therapy. CONCLUSION: By explicitly modelling the underlying abrupt changes in mortality risk after initiation of antiretroviral therapy we are able to estimate their number and location in a rigorous, data-driven manner. The existence of a high early risk of death after initiation of antiretroviral therapy and the determination of its duration has direct implications for the optimal management of patients initiating therapy in this setting. PMID- 19558674 TI - Genome analysis and genome-wide proteomics of Thermococcus gammatolerans, the most radioresistant organism known amongst the Archaea. AB - BACKGROUND: Thermococcus gammatolerans was isolated from samples collected from hydrothermal chimneys. It is one of the most radioresistant organisms known amongst the Archaea. We report the determination and annotation of its complete genome sequence, its comparison with other Thermococcales genomes, and a proteomic analysis. RESULTS: T. gammatolerans has a circular chromosome of 2.045 Mbp without any extra-chromosomal elements, coding for 2,157 proteins. A thorough comparative genomics analysis revealed important but unsuspected genome plasticity differences between sequenced Thermococcus and Pyrococcus species that could not be attributed to the presence of specific mobile elements. Two virus related regions, tgv1 and tgv2, are the only mobile elements identified in this genome. A proteogenome analysis was performed by a shotgun liquid chromatography tandem mass spectrometry approach, allowing the identification of 10,931 unique peptides corresponding to 951 proteins. This information concurrently validates the accuracy of the genome annotation. Semi-quantification of proteins by spectral count was done on exponential- and stationary-phase cells. Insights into general catabolism, hydrogenase complexes, detoxification systems, and the DNA repair toolbox of this archaeon are revealed through this genome and proteome analysis. CONCLUSIONS: This work is the first archaeal proteome investigation done at the stage of primary genome annotation. This archaeon is shown to use a large variety of metabolic pathways even under a rich medium growth condition. This proteogenomic study also indicates that the high radiotolerance of T. gammatolerans is probably due to proteins that remain to be characterized rather than a larger arsenal of known DNA repair enzymes. PMID- 19558678 TI - Retention of the virus-derived sequences in the nuclear genome of grapevine as a potential pathway to virus resistance. AB - BACKGROUND: Previous studies have revealed a wide-spread occurence of the partial and complete genomes of the reverse-transcribing pararetroviruses in the nuclear genomes of herbaceous plants. Although the absence of the virus-encoded integrases attests to the random and incidental incorporation of the viral sequences, their presence could have functional implications for the virus-host interactions. HYPOTHESIS: Analyses of two nuclear genomes of grapevine revealed multiple events of horizontal gene transfer from pararetroviruses. The approximately 200-800 bp inserts that corresponded to partial ORFs encoding reverse transcriptase apparently derived from unknown or extinct caulimoviruses and tungroviruses, were found in 11 grapevine chromosomes. In contrast to the previous reports, no reliable cases of the inserts derived from the positive strand RNA viruses were found. Because grapevine is known to be infected by the diverse positive-strand RNA viruses, but not pararetroviruses, we hypothesize that pararetroviral inserts have conferred host resistance to these viruses. Furthermore, we propose that such resistance involves RNA interference-related mechanisms acting via small RNA-mediated methylation of pararetroviral DNAs and/or via degradation of the viral mRNAs. CONCLUSION: The pararetroviral sequences in plant genomes may be maintained due to the benefits of virus resistance to this class of viruses conferred by their presence. Such resistance could be particularly significant for the woody plants that must withstand years- to centuries-long virus assault. Experimental research into the RNA interference pathways involving the integrated pararetroviral inserts is required to test this hypothesis. REVIEWERS: This article was reviewed by Arcady R. Mushegian, I. King Jordan, and Eugene V. Koonin. PMID- 19558679 TI - Antimicrobial breakpoint estimation accounting for variability in pharmacokinetics. AB - BACKGROUND: Pharmacokinetic and pharmacodynamic (PK/PD) indices are increasingly being used in the microbiological field to assess the efficacy of a dosing regimen. In contrast to methods using MIC, PK/PD-based methods reflect in vivo conditions and are more predictive of efficacy. Unfortunately, they entail the use of one PK-derived value such as AUC or Cmax and may thus lead to biased efficiency information when the variability is large. The aim of the present work was to evaluate the efficacy of a treatment by adjusting classical breakpoint estimation methods to the situation of variable PK profiles. METHODS AND RESULTS: We propose a logical generalisation of the usual AUC methods by introducing the concept of "efficiency" for a PK profile, which involves the efficacy function as a weight. We formulated these methods for both classes of concentration- and time dependent antibiotics. Using drug models and in silico approaches, we provide a theoretical basis for characterizing the efficiency of a PK profile under in vivo conditions. We also used the particular case of variable drug intake to assess the effect of the variable PK profiles generated and to analyse the implications for breakpoint estimation. CONCLUSION: Compared to traditional methods, our weighted AUC approach gives a more powerful PK/PD link and reveals, through examples, interesting issues about the uniqueness of therapeutic outcome indices and antibiotic resistance problems. PMID- 19558680 TI - Effectiveness of a tinnitus management programme: a 2-year follow-up study. AB - BACKGROUND: Tinnitus impairs the possibility of leading a normal life in 0.5-1% of the population. While neither medical nor surgical treatment appears effective, counselling may offer some relief. An intervention combining counselling and hearing devices is offered to clients referred to the Centre for Help Aids and Communication (CHC) in southern Denmark. The aims of this exploratory study were to examine i) the characteristics of CHC's clients and their tinnitus, ii) the effectiveness of the treatment, and iii) whether particular client groups benefit more than others. METHODS: One hundred new clients presenting with tinnitus completed the Tinnitus Handicap Inventory (THI) three times - before their first consultation, after one month and after 1-2 years. The scores were tested for significant differences over time using tests for paired data. Logistic regression was used to examine factors associated with a clinically important difference (i.e. THI score improvement of at least 20 points). RESULTS: At final follow-up, total THI score was significantly lower than baseline, i.e. 29.8 (CI 25.5-34.2) vs. 37.2 (CI 33.1-37.2), p < 0.01. The programme achieved a clinically important difference for 27% and 24% of the clients one month and 1-2 years after the first consultation, respectively. It appeared that greater improvement in THI score was related to higher baseline THI score and possibly also to treatment by a particular CHC therapist. The absolute reduction in mean THI score after 1-2 years for clients with moderate and severe handicap was 14 and 20 points, respectively, i.e. similar to that previously reported for TRT (14-28 points). The cost of the current programme was approximately 200 EUR per client. CONCLUSION: The tinnitus management programme appeared to provide significant benefit to many clients at a relatively low cost. It would be useful to conduct a randomised controlled study comparing the current programme with alternative forms of combination counselling/sound therapy approaches. PMID- 19558681 TI - Antifungal treatment for invasive Candida infections: a mixed treatment comparison meta-analysis. AB - OBJECTIVES: Invasive fungal infections are a major cause of mortality among patients at risk. Treatment guidelines vary on optimal treatment strategies. We aimed to determine the effects of different antifungal therapies on global response rates, mortality and safety. METHODS: We searched independently and in duplicate 10 electronic databases from inception to May 2009. We selected any randomized trial assessing established antifungal therapies for confirmed cases of invasive candidiasis among predominantly adult populations. We performed a meta-analysis and then conducted a Bayesian mixed treatment comparison to differentiate treatment effectiveness. Sensitivity analyses included dosage forms of amphotericin B and fluconazole compared to other azoles. RESULTS: Our analysis included 11 studies enrolling a total of 965 patients. For our primary analysis of global response rates, we pooled 7 trials comparing azoles to amphotericin B, Relative Risk [RR] 0.87 (95% Confidence Interval [CI], 0.78-0.96, P = 0.007, I2 = 43%, P = 0.09. We also pooled 2 trials of echinocandins versus amphotericin B and found a pooled RR of 1.10 (95% CI, 0.99-1.23, P = 0.08). One study compared anidulafungin to fluconazole and yielded a RR of 1.26 (95% CI, 1.06-1.51) in favor of anidulafungin. We pooled 7 trials assessing azoles versus amphotericin B for all-cause mortality, resulting in a pooled RR of 0.88 (95% CI, 0.74-1.05, P = 0.17, I2 = 0%, P = 0.96). Echinocandins versus amphotericin B (2 trials) for all cause mortality resulted in a pooled RR of 1.01 (95% CI, 0.84-1.20, P = 0.93). Anidulafungin versus fluconazole resulted in a RR of 0.73 (95% CI, 0.48-1.10, P = 0.34). Our mixed treatment comparison analysis found similar within-class effects across all interventions. Adverse event profiles differed, with amphotericin B exhibiting larger adverse event effects. CONCLUSION: Treatment options appear to offer preferential effects on response rates and mortality. When mycologic data are available, therapy should be tailored. PMID- 19558682 TI - Designing financial-incentive programmes for return of medical service in underserved areas: seven management functions. AB - In many countries worldwide, health worker shortages are one of the main constraints in achieving population health goals. Financial-incentive programmes for return of service, whereby participants receive payments in return for a commitment to practise for a period of time in a medically underserved area, can alleviate local and regional health worker shortages through a number of mechanisms. First, they can redirect the flow of those health workers who would have been educated without financial incentives from well-served to underserved areas. Second, they can add health workers to the pool of workers who would have been educated without financial incentives and place them in underserved areas. Third, financial-incentive programmes may improve the retention in underserved areas of those health workers who participate in a programme, but who would have worked in an underserved area without any financial incentives. Fourth, the programmes may increase the retention of all health workers in underserved areas by reducing the strength of some of the reasons why health workers leave such areas, including social isolation, lack of contact with colleagues, lack of support from medical specialists and heavy workload. We draw on studies of financial-incentive programmes and other initiatives with similar objectives to discuss seven management functions that are essential for the long-term success of financial-incentive programmes: financing (programmes may benefit from innovative donor financing schemes, such as endowment funds, international financing facilities or compensation payments); promotion (programmes should use tested communication channels in order to reach secondary school graduates and health workers); selection (programmes may use selection criteria to ensure programme success and to achieve supplementary policy goals); placement (programmes should match participants to areas in order to maximize participant satisfaction and retention); support (programmes should prepare participants for the time in an underserved area, stay in close contact with participants throughout the different phases of enrolment and help participants by assigning them mentors, establishing peer support systems or financing education courses relevant to work in underserved areas); enforcement (programmes may use community based monitoring or outsource enforcement to existing institutions); and evaluation (in order to broaden the evidence on the effectiveness of financial incentives in increasing the health workforce in underserved areas, programmes in developing countries should evaluate their performance; in order to improve the strength of the evidence on the effectiveness of financial incentives, controlled experiments should be conducted where feasible). In comparison to other interventions to increase the supply of health workers to medically underserved areas, financial-incentive programmes have advantages--unlike initiatives using non-financial incentives, they establish legally enforceable commitments to work in underserved areas and, unlike compulsory service policies, they will not be opposed by health workers--as well as disadvantages--unlike initiatives using non financial incentives, they may not improve the working and living conditions in underserved areas (which are important determinants of health workers' long-term retention) and, unlike compulsory service policies, they cannot guarantee that they will supply health workers to underserved areas who would not have worked in such areas without financial incentives. Financial incentives, non-financial incentives, and compulsory service are not mutually exclusive and may positively affect each other's performance. PMID- 19558683 TI - Neurodegeneration in Alzheimer's disease: caspases and synaptic element interdependence. AB - Extensive genetic, biochemical, and histological evidence has implicated the amyloid-beta peptide (Abeta) in Alzheimer's disease pathogenesis, and several mechanisms have been suggested, such as metal binding, reactive oxygen species production, and membrane pore formation. However, recent evidence argues for an additional role for signaling mediated by the amyloid precursor protein, APP, in part via the caspase cleavage of APP at aspartate 664. Here we review the effects and implications of this cleavage event, and propose a model of Alzheimer's disease that focuses on the critical nature of this cleavage and its downstream effects. PMID- 19558684 TI - Structural and functional implications of p53 missense cancer mutations. AB - Most human cancers contain mutations in the transcription factor p53 and majority of these are missense and located in the DNA binding core domain. In this study, the stabilities of all core domain missense mutations are predicted and are used to infer their likely inactivation mechanisms. Overall, 47.0% non PRO/GLY mutants are stable (DeltaDeltaG < 1.0 kT) and 36.3% mutants are unstable (DeltaDeltaG > 3.0 kT), 12.2% mutants are with 1.0 kT < DeltaDeltaG < 3.0 kT. Only 4.5% mutants are with no conclusive predictions. Certain types of either stable or unstable mutations are found not to depend on their local structures. Y, I, C, V, F and W (W, R and F) are the most common residues before (after) mutation in unstable mutants. Q, N, K, D, A, S and T (I, T, L and V) are the most common residues before (after) mutation in stable mutants. The stability correlations with sequence, structure, and molecular contacts are also analyzed. No direct correlation between secondary structure and stability is apparent, but a strong correlation between solvent exposure and stability is noticeable. Our correlation analysis shows that loss of protein-protein contacts may be an alternative cause for p53 inactivation. Correlation with clinical data shows that loss of stability and loss of DNA contacts are the two main inactivation mechanisms. Finally, correlation with functional data shows that most mutations which retain functions are stable, and most mutations that gain functions are unstable, indicating destabilized and deformed p53 proteins are more likely to find new binding partners.PACS codes: 87.14.E- PMID- 19558685 TI - "How I do it: utilization of high-pressure sealants in aortic reconstruction". AB - BACKGROUND: Suture-line hemostasis, reinforcement of friable tissue, and adhesion prevention are key concerns for patients undergoing cardiac surgery for aortic reconstruction. Failure to secure hemostasis at anastomotic junctures and reinforce fragile tissue may lead to increased blood loss, additional blood product requirements, increased operative time, and, in extreme cases, reoperation. Patients with aortic pathology may also be at higher risk for reoperation, and adhesion formation from prior surgery is an added risk at resternotomy. The advent of high-pressure sealants has been of benefit in helping to alleviate these perioperative challenges. METHODS: The author utilizes two high-pressure sealants for aortic reconstructive procedures. The first is made of two polymers of polyethylene glycol (PEG) [Coseal, Baxter Healthcare, Corporation], and is used to secure anastomotic suture-line hemostasis and for adhesion prevention. The second is a bovine serum albumin-glutaraldehyde (BSAG) glue [BioGlue, CryroLife, Inc.], used for the repair of dissected aortic tissue and in reinforcing ("tanning") fragile aortic tissues. The techniques for application in select aortic reconstruction procedures are described. RESULTS: To substantiate the hemostatic clinical benefit observed by the author, 60 consecutive major thoracic aortic operations in 57 patients in whom PEG sealant was used were retrospectively reviewed. Although comparisons with other agents were not performed for this descriptive report, bleeding results were very favorable for these types of operations. The strong clinical impression is that topical hemostatic application of PEG sealant to anastomotic suture lines is helpful in preventing bleeding. CONCLUSION: In major aortic reconstructive procedures the need for anastomotic sealing performance, reinforcement of friable tissues, and adhesion prevention should not be underrated. High-pressure surgical sealants represent an important surgical adjunct, and the author has found the use of both PEG sealant and BSAG glue advantageous in aortic reconstruction and repair. PMID- 19558686 TI - Is there a relationship between factor V Leiden and type 2 diabetes? AB - BACKGROUND: Diabetes is well known risk factor for thrombotic events. The association between diabetes and venous thromboembolism is still matter of debate. However, during diabetes an acquired thrombophilia is present and is due to the non-enzymatic glycosilation of clotting inhibitors as antithrombin thus leading to hypercoagulable state. A possibile relationship between the presence of FVL gene variant in type 1 or type 2 diabetes has been hypothysed by several reports in the Literature with non-univocal findings. PATIENTS AND METHODS: Retrospectively we analysed nearly 7000 patients referred to our Thrombosis Center for venous thromboembolism (VTE) then we selected 115 patients underwent to the screening for inherited thrombophilia. All selected patients were divided in 2 groups: the first group (group A) included 64 patients with previous VTE and carriers of factor V Leiden, while the second group (group B) included 51 patients with previous VTE and evetually carriers of thrombophilic defects other than factor V Leiden. Patients of group B acted as control group. 75 g oral glucose tolerance Test (OGTT) recommended by WHO was perfomed to all subjects in the study in order to screen subjects with glucose reduced tolerance or subjects with inducible diabetes. Statistical analysis was performed with STATA 6 http://www.stata.com with Student t test for unpaired data, with chi2 test or with Fisher exact test where appropriated; differences were considered to be significant if p < 0.05. RESULTS: We did not find sifferences between glycaemia at baseline and after OGTT between patients with VTE carriers of FVL compared to non-carriers of FVL. We found a relevant increase in the prevalence of IGT and diabetes between patients with VTE carriers of FVL compared to non-carriers of FVL although this increase did not raise statistical significance. DISCUSSION: our data pointed out an interesting aspect of the linking between FVL gene variant, diabetes and atherothrombosis and other vascular complications, although data on larger population are needed; this aspect may be another relevant topic of research based because also a link between the pathogenesis of venous thrombosis and atherothrombosis has been recently reported in the Literature. PMID- 19558687 TI - Tularemia induces different biochemical responses in BALB/c mice and common voles. AB - BACKGROUND: Both BALB/c mice and common voles (Microtus arvalis) are considered highly susceptible to tularemia. However, the common vole is reported to harbour Francisella tularensis in European habitats as well as to survive longer with chronic shedding of the bacterium. The purpose of the present study was to compare the response of these two rodents to a wild Francisella tularensis subsp. holarctica strain infection. METHODS: Rodents were evaluated for differences in the total antioxidant capacity derived from low-molecular-weight antioxidants, biochemistry including lipid metabolism, tissue bacterial burdens and histopathology following experimental intraperitoneal infection with 160 colony forming units (CFU) pro toto. RESULTS: Bacterial burdens in common voles started to develop later post-exposure and amounted to lower levels than in BALB/c mice. Elevation of liver function enzymes was more pronounced in mice than common voles and there were marked differences in lipid metabolism in the course of tularemia in these two species. Hypertriglyceridemia and hypercholesterolemia developed in mice, while physiologically higher levels of triglycerides and cholesterol showed a decreasing tendency in common voles. On the other hand, the total plasma antioxidant capacity gradually dropped to 81.5% in mice on day 5 post-infection, while it increased to 130% on day 6 post-infection in common voles. Significant correlations between tissue bacterial burdens and several biochemical parameters were found. CONCLUSION: As differences in lipid metabolism and the total antioxidant capacity of highly susceptible rodent species were demonstrated, the role of triglycerides, cholesterol and antioxidants in tularemic sepsis should be further investigated. PMID- 19558688 TI - Antimicrobial activities of commercial nanoparticles against an environmental soil microbe, Pseudomonas putida KT2440. AB - BACKGROUND: The release of heavy metal-containing nanoparticles (NP) into the environment may be harmful to the efficacy of beneficial microbes that function in element cycling, pollutant degradation and plant growth. Nanoparticles of Ag, CuO and ZnO are of interest as antimicrobials against pathogenic bacteria. We demonstrate here their antimicrobial activity against the beneficial soil microbe, Pseudomonas putida KT2440. RESULTS: Toxicity was detected in a KT2440 construct possessing a plasmid bearing the luxAB reporter genes. "As manufactured" preparations of nano- Ag, -CuO and -ZnO caused rapid dose-dependent loss of light output in the biosensor. Cell death accompanied loss in Lux activity with treatments by nano-Ag and -CuO, but with -ZnO the treatments were bacteriostatic rather than bactericidal. Bulk equivalents of these products showed no inhibitory activity, indicating that particle size was determinant in activity. Flow Field-Flow Fractionation (FlFFF) of an aqueous suspension of the nano-CuO and ZnO revealed a small proportion of 5 nm NP and aggregated particulates with sizes ranging between 70 nm and 300 nm; the majority portion of material was aggregated into particles larger than 300 nm in size. Thus within the commercial preparation there may be microbially active and inactive forms. CONCLUSION: The "as-made" NP of Ag, CuO and ZnO have toxic effects on a beneficial soil microbe, leading to bactericidal or bacteriostatic effects depending on the NP employed. The lack of toxicity from bulk materials suggests that aggregation of the NP into larger particles, possibly by factors present in the environment may reduce their nontarget antimicrobial activity. PMID- 19558690 TI - Leiomyosarcoma of the inferior vena cava: radical surgery and vascular reconstruction. AB - BACKGROUND: Vascular leiomyosarcoma are rare tumors typically originating from the inferior vena cava (IVC). Due to nonspecific clinical signs most tumors are diagnosed at advanced stages. Complete surgical resection remains the only potential curative therapeutic option. Surgical strategy is particularly influenced by the level of the IVC affected. Due to the topographic relation to the renal veins level-II involvement of the IVC raises special surgical challenges with respect to the maintenance of venous outflow. CASE PRESENTATION: We herein report two cases of leiomyosarcoma of the IVC with successful en bloc resection and individualized caval reconstruction. One patient presented with a large intramural and intraluminal mass and received a complete circumferential resection. Reconstruction was performed by graft replacement of the caval segment affected. The other patient displayed a predominantly extraluminal tumor growth and underwent semicircumferential resection of the IVC including the confluence of the left renal vein. In this case vascular reconstruction was performed by cavoplasty and reinsertion of the left renal vein into the proximal portion of the IVC. Resection margins of both patients were tumor free and no clinical signs of venous insufficiency of the lower extremity occurred. CONCLUSION: This paper presents two cases of successfully managed leiomyosarcomas of the vena cava and exemplifies two different options for vascular reconstruction in level II sarcomas and includes a thorough review of the literature. PMID- 19558689 TI - Contextual blending of ingroup/outgroup face stimuli and word valence: LPP modulation and convergence of measures. AB - BACKGROUND: Several event related potential (ERP) studies have investigated the time course of different aspects of evaluative processing in social bias research. Various reports suggest that the late positive potential (LPP) is modulated by basic evaluative processes, and some reports suggest that in /outgroup relative position affects ERP responses. In order to study possible LPP blending between facial race processing and semantic valence (positive or negative words), we recorded ERPs while indigenous and non-indigenous participants who were matched by age and gender performed an implicit association test (IAT). The task involved categorizing faces (ingroup and outgroup) and words (positive and negative). Since our paradigm implies an evaluative task with positive and negative valence association, a frontal distribution of LPPs similar to that found in previous reports was expected. At the same time, we predicted that LPP valence lateralization would be modulated not only by positive/negative associations but also by particular combinations of valence, face stimuli and participant relative position. RESULTS: Results showed that, during an IAT, indigenous participants with greater behavioral ingroup bias displayed a frontal LPP that was modulated in terms of complex contextual associations involving ethnic group and valence. The LPP was lateralized to the right for negative valence stimuli and to the left for positive valence stimuli. This valence lateralization was influenced by the combination of valence and membership type relevant to compatibility with prejudice toward a minority. Behavioral data from the IAT and an explicit attitudes questionnaire were used to clarify this finding and showed that ingroup bias plays an important role. Both ingroup favoritism and indigenous/non-indigenous differences were consistently present in the data. CONCLUSION: Our results suggest that frontal LPP is elicited by contextual blending of evaluative judgments of in-/outgroup information and positive vs. negative valence association and confirm recent research relating in-/outgroup ERP modulation and frontal LPP. LPP modulation may cohere with implicit measures of attitudes. The convergence of measures that were observed supports the idea that racial and valence evaluations are strongly influenced by context. This result adds to a growing set of evidence concerning contextual sensitivity of different measures of prejudice. PMID- 19558691 TI - CHOP 5'UTR-c.279T>C and +nt30C>T variants are not associated with overweight condition or with tumors/cancer in Italians - a case-control study. AB - BACKGROUND: Type 2 diabetes (T2D) is associated with obesity and has been shown recently to be associated with tumors/cancer. HNF1-beta and JAZF1 genes are associated with T2D and prostate cancer. We have previously shown that CHOP 5'UTR c.279T>C and +nt30C>T haplotype variants contribute to T2D. CHOP deficiency causes obesity in mice, thus CHOP gene variants may contribute to human obesity. Furthermore, CHOP mediates apoptosis and is implicated in cancer pathogenesis. Hence, we aimed at identifying any potential association of CHOP 5'UTR-c.279T>C and +nt30C>T genotypes and corresponding haplotypes with overweight condition/pre obesity and tumors/cancer in an Italian dataset. METHODS: We recruited from Italy 45 overweight subjects (body mass index (BMI) >or= 25) and 44 control subjects (BMI < 25) as well as 54 cases with at least one cancer or at least one tumor and 43 control subjects without tumors/cancer from the general population. We excluded allelic departure from Hardy-Weinberg equilibrium in cases and control subjects, separately. RESULTS: We assessed the power to detect risk odds ratios by association tests in our datasets. We tested the hypothesis of association of CHOP 5'UTR-c.279T>C and +nt30C>T genotypes and haplotypes with tumors/cancer and, separately, with overweight condition. Both associations were not significant. CONCLUSION: From our study, we may conclude that CHOP 5'UTR-c.279T>C and +nt30C>T genotypes and corresponding haplotypes are not associated with tumors/cancer and pre-obesity. However, more studies are warranted to establish the role of CHOP variants in tumor/cancer predisposition and in overweight condition. PMID- 19558692 TI - Altered regulation of metabolic pathways in human lung cancer discerned by (13)C stable isotope-resolved metabolomics (SIRM). AB - BACKGROUND: Metabolic perturbations arising from malignant transformation have not been systematically characterized in human lung cancers in situ. Stable isotope resolved metabolomic analysis (SIRM) enables functional analysis of gene dysregulations in lung cancer. To this purpose, metabolic changes were investigated by infusing uniformly labeled 13C-glucose into human lung cancer patients, followed by resection and processing of paired non-cancerous lung and non small cell carcinoma tissues. NMR and GC-MS were used for 13C-isotopomer based metabolomic analysis of the extracts of tissues and blood plasma. RESULTS: Many primary metabolites were consistently found at higher levels in lung cancer tissues than their surrounding non-cancerous tissues. 13C-enrichment in lactate, Ala, succinate, Glu, Asp, and citrate was also higher in the tumors, suggesting more active glycolysis and Krebs cycle in the tumor tissues. Particularly notable were the enhanced production of the Asp isotopomer with three 13C-labeled carbons and the buildup of 13C-2,3-Glu isotopomer in lung tumor tissues. This is consistent with the transformations of glucose into Asp or Glu via glycolysis, anaplerotic pyruvate carboxylation (PC), and the Krebs cycle. PC activation in tumor tissues was also shown by an increased level of pyruvate carboxylase mRNA and protein. CONCLUSION: PC activation - revealed here for the first time in human subjects - may be important for replenishing the Krebs cycle intermediates which can be diverted to lipid, protein, and nucleic acid biosynthesis to fulfill the high anabolic demands for growth in lung tumor tissues. We hypothesize that this is an important event in non-small cell lung cancer and possibly in other tumor development. PMID- 19558693 TI - Proneoplastic effects of PGE2 mediated by EP4 receptor in colorectal cancer. AB - BACKGROUND: Prostaglandin E2 (PGE2) is the major product of Cyclooxygenase-2 (COX 2) in colorectal cancer (CRC). We aimed to assess PGE2 cell surface receptors (EP 1-4) to examine the mechanisms by which PGE2 regulates tumour progression. METHODS: Gene expression studies were performed by quantitative RT-PCR. Cell cycle was analysed by flow cytometry with cell proliferation quantified by BrdU incorporation measured by enzyme immunoassay. Immunohistochemistry was employed for expression studies on formalin fixed paraffin embedded tumour tissue. RESULTS: EP4 was the most abundant subtype of PGE2 receptor in HT-29 and HCA7 cells (which show COX-2 dependent PGE2 generation) and was consistently the most abundant transcript in human colorectal tumours (n = 8) by qRT-PCR (ANOVA, p = 0.01). G0/G1 cell cycle arrest was observed in HT-29 cells treated with SC-236 5 microM (selective COX-2 inhibitor) for 24 hours (p = 0.02), an effect abrogated by co-incubation with PGE2 (1 microM). G0/G1 arrest was also seen with a specific EP4 receptor antagonist (EP4A, L-161982) (p = 0.01). Treatment of HT-29 cells with either SC-236 or EP4A caused reduction in intracellular cAMP (ANOVA, p = 0.01). Early induction in p21WAF1/CIP1 expression (by qRT-PCR) was seen with EP4A treatment (mean fold increase 4.4, p = 0.04) while other genes remained unchanged. Similar induction in p21WAF1/CIP1 was also seen with PD153025 (1 microM), an EGFR tyrosine kinase inhibitor, suggesting EGFR transactivation by EP4 as a potential mechanism. Additive inhibition of HCA7 proliferation was observed with the combination of SC-236 and neutralising antibody to amphiregulin (AR), a soluble EGFR ligand. Concordance in COX-2 and AR localisation in human colorectal tumours was noted. CONCLUSION: COX-2 regulates cell cycle transition via EP4 receptor and altered p21WAF1/CIP1 expression. EGFR pathways appear important. Specific targeting of the EP4 receptor or downstream targets may offer a safer alternative to COX-2 inhibition in the chemoprevention of CRC. PMID- 19558695 TI - The spatial and temporal patterns of falciparum and vivax malaria in Peru: 1994 2006. AB - BACKGROUND: Malaria is the direct cause of approximately one million deaths worldwide each year, though it is both preventable and curable. Increasing the understanding of the transmission dynamics of falciparum and vivax malaria and their relationship could suggest improvements for malaria control efforts. Here the weekly number of malaria cases due to Plasmodium falciparum (1994-2006) and Plasmodium vivax (1999-2006) in Peru at different spatial scales in conjunction with associated demographic, geographic and climatological data are analysed. METHODS: Malaria periodicity patterns were analysed through wavelet spectral analysis, studied patterns of persistence as a function of community size and assessed spatial heterogeneity via the Lorenz curve and the summary Gini index. RESULTS: Wavelet time series analyses identified annual cycles in the incidence of both malaria species as the dominant pattern. However, significant spatial heterogeneity was observed across jungle, mountain and coastal regions with slightly higher levels of spatial heterogeneity for P. vivax than P. falciparum. While the incidence of P. falciparum has been declining in recent years across geographic regions, P. vivax incidence has remained relatively steady in jungle and mountain regions with a slight decline in coastal regions. Factors that may be contributing to this decline are discussed. The time series of both malaria species were significantly synchronized in coastal (rho = 0.9, P < 0.0001) and jungle regions (rho = 0.76, P < 0.0001) but not in mountain regions. Community size was significantly associated with malaria persistence due to both species in jungle regions, but not in coastal and mountain regions. CONCLUSION: Overall, findings highlight the importance of highly refined spatial and temporal data on malaria incidence together with demographic and geographic information in improving the understanding of malaria persistence patterns associated with multiple malaria species in human populations, impact of interventions, detection of heterogeneity and generation of hypotheses. PMID- 19558696 TI - On the road to diploidization? Homoeolog loss in independently formed populations of the allopolyploid Tragopogon miscellus (Asteraceae). AB - BACKGROUND: Polyploidy (whole-genome duplication) is an important speciation mechanism, particularly in plants. Gene loss, silencing, and the formation of novel gene complexes are some of the consequences that the new polyploid genome may experience. Despite the recurrent nature of polyploidy, little is known about the genomic outcome of independent polyploidization events. Here, we analyze the fate of genes duplicated by polyploidy (homoeologs) in multiple individuals from ten natural populations of Tragopogon miscellus (Asteraceae), all of which formed independently from T. dubius and T. pratensis less than 80 years ago. RESULTS: Of the 13 loci analyzed in 84 T. miscellus individuals, 11 showed loss of at least one parental homoeolog in the young allopolyploids. Two loci were retained in duplicate for all polyploid individuals included in this study. Nearly half (48%) of the individuals examined lost a homoeolog of at least one locus, with several individuals showing loss at more than one locus. Patterns of loss were stochastic among individuals from the independently formed populations, except that the T. dubius copy was lost twice as often as T. pratensis. CONCLUSION: This study represents the most extensive survey of the fate of genes duplicated by allopolyploidy in individuals from natural populations. Our results indicate that the road to genome downsizing and ultimate genetic diploidization may occur quickly through homoeolog loss, but with some genes consistently maintained as duplicates. Other genes consistently show evidence of homoeolog loss, suggesting repetitive aspects to polyploid genome evolution. PMID- 19558697 TI - Development and validation of a HPV-32 specific PCR assay. AB - BACKGROUND: Human Papillomavirus-32 (HPV-32) has traditionally been associated with focal-epithelial-hyperplasia (FEH). It is also present in 58% of oral warts of HIV-positive individuals whose prevalence is increasing. Current methods for the detection of HPV-32 are labor-intensive and insensitive so the goal of this work was to develop a highly sensitive and easy to use specific polymerase chain reaction (PCR) assay. MATERIALS AND METHODS: An HPV-32 L1 specific PCR assay was developed and optimized. The sensitivity and specificity was compared to previous assays utilized for detection (PGMY and MY09/11 PCR with dot blot hybridization) using cloned HPV-32 L1, the closely related HPV-42 L1 as well as clinical samples (oral swabs and fluids from 89 HIV-positive subjects). RESULTS: The HPV-32 specific PCR assay showed improved sensitivity to 5 copies of HPV-32 as compared to the PGMY PCR, MY09/11 PCR and dot blot which had a limit of detection of approximately 3,000 copies. Using the HPV-32 dot blot hybridization assay as the gold standard, the HPV-32 specific PCR assay has a sensitivity of 95.8% and 88.9% by sample and subject, respectively, and specificity was 87.8% and 58.8% by sample and subject, respectively. The low sensitivity is due to the HPV-32 specific PCR assays ability to detect more HPV-32 positive samples and may be the new gold standard. CONCLUSION: Due to the ease, sensitivity, and specificity the HPV-32 specific PCR assay is superior to previous assays and is ideal for detection of HPV-32 in large cohorts. This assay provides an excellent tool to study the natural history of HPV-32 infection and the development of oral warts. PMID- 19558694 TI - Triangle network motifs predict complexes by complementing high-error interactomes with structural information. AB - BACKGROUND: A lot of high-throughput studies produce protein-protein interaction networks (PPINs) with many errors and missing information. Even for genome-wide approaches, there is often a low overlap between PPINs produced by different studies. Second-level neighbors separated by two protein-protein interactions (PPIs) were previously used for predicting protein function and finding complexes in high-error PPINs. We retrieve second level neighbors in PPINs, and complement these with structural domain-domain interactions (SDDIs) representing binding evidence on proteins, forming PPI-SDDI-PPI triangles. RESULTS: We find low overlap between PPINs, SDDIs and known complexes, all well below 10%. We evaluate the overlap of PPI-SDDI-PPI triangles with known complexes from Munich Information center for Protein Sequences (MIPS). PPI-SDDI-PPI triangles have ~20 times higher overlap with MIPS complexes than using second-level neighbors in PPINs without SDDIs. The biological interpretation for triangles is that a SDDI causes two proteins to be observed with common interaction partners in high throughput experiments. The relatively few SDDIs overlapping with PPINs are part of highly connected SDDI components, and are more likely to be detected in experimental studies. We demonstrate the utility of PPI-SDDI-PPI triangles by reconstructing myosin-actin processes in the nucleus, cytoplasm, and cytoskeleton, which were not obvious in the original PPIN. Using other complementary datatypes in place of SDDIs to form triangles, such as PubMed co occurrences or threading information, results in a similar ability to find protein complexes. CONCLUSION: Given high-error PPINs with missing information, triangles of mixed datatypes are a promising direction for finding protein complexes. Integrating PPINs with SDDIs improves finding complexes. Structural SDDIs partially explain the high functional similarity of second-level neighbors in PPINs. We estimate that relatively little structural information would be sufficient for finding complexes involving most of the proteins and interactions in a typical PPIN. PMID- 19558698 TI - Discovery of novel inhibitors of Streptococcus pneumoniae based on the virtual screening with the homology-modeled structure of histidine kinase (VicK). AB - BACKGROUND: Due to the widespread abusage of antibiotics, antibiotic-resistance in Streptococcus pneumoniae (S. pneumoniae) has been increasing quickly in recent years, and it is obviously urgent to develop new types of antibiotics. Two component systems (TCSs) are the major signal transduction pathways in bacteria and have emerged as potential targets for antibacterial drugs. Among the 13 pairs of TCSs proteins presenting in S. pneumoniae, VicR/K is the unique one essential for bacterium growth, and block agents to which, if can be found, may be developed as effective antibiotics against S. pneumoniae infection. RESULTS: Using a structure-based virtual screening (SBVS) method, 105 compounds were computationally identified as potential inhibitors of the histidine kinase (HK) VicK protein from the compound library SPECS. Six of them were then validated in vitro to be active in inhibiting the growth of S. pneumoniae without obvious cytotoxicity to Vero cell. In mouse sepsis models, these compounds are still able to decrease the mortality of the mice infected by S. pneumoniae and one compound even has significant therapeutic effect. CONCLUSION: To our knowledge, these compounds are the first reported inhibitors of HK with antibacterial activity in vitro and in vivo, and are novel lead structures for developing new drugs to combat pneumococcal infection. PMID- 19558700 TI - Prevalence of hypospadias in Italy according to severity, gestational age and birthweight: an epidemiological study. AB - BACKGROUND: Hypospadias is a congenital displacement of the urethral meatus in male newborns, being either an isolated defect at birth or a sign of sexual development disorders. The aim of this study was to assess the prevalence rate of hypospadias in different Districts of Italy, in order to make a comparison with other countries all over the world. METHODS: We reviewed all the newborns file records (years 2001-2004) in 15 Italian Hospitals. RESULTS: We found an overall hypospadias prevalence rate of 3.066 +/- 0.99 per 1000 live births (82.48% mild hypospadias, 17.52% moderate-severe). In newborns Small for Gestational Age (birthweight < 10(th )percentile) of any gestational age the prevalence rate of hypospadias was 6.25 per 1000 live births. Performing multivariate logistic regression analysis for different degrees of hypospadias according to severity, being born SGA remained the only risk factor for moderate-severe hypospadias (p = 0.00898) but not for mild forms (p > 0.1). CONCLUSION: In our sample the prevalence of hypospadias results as high as reported in previous European and American studies (3-4 per 1000 live births). Pathogenesis of isolated hypospadias is multifactorial (genetic, endocrine and environmental factors): however, the prevalence rate of hypospadias is higher in infants born small for gestational age than in newborns with normal birth weight. PMID- 19558699 TI - Phylogeography of the Alcippe morrisonia (Aves: Timaliidae): long population history beyond late Pleistocene glaciations. AB - BACKGROUND: The role of Pleistocene glacial oscillations in current biodiversity and distribution patterns varies with latitude, physical topology and population life history and has long been a topic of discussion. However, there had been little phylogeographical research in south China, where the geophysical complexity is associated with great biodiversity. A bird endemic in Southeast Asia, the Grey-cheeked Fulvetta, Alcippe morrisonia, has been reported to show deep genetic divergences among its seven subspecies. In the present study, we investigated the phylogeography of A. morrisonia to explore its population structure and evolutionary history, in order to gain insight into the effect of geological events on the speciation and diversity of birds endemic in south China. RESULTS: Mitochondrial genes cytochrome b (Cytb) and cytochrome c oxidase I (COI) were represented by 1236 nucleotide sites from 151 individuals from 29 localities. Phylogenetic analysis showed seven monophyletic clades congruent with the geographically separated groups, which were identified as major sources of molecular variance (90.92%) by AMOVA. TCS analysis revealed four disconnected networks, and that no haplotype was shared among the geographical groups. The common ancestor of these populations was dated to 11.6 Mya and several divergence events were estimated along the population evolutionary history. Isolation by distance was inferred by NCPA to be responsible for the current intra-population genetic pattern and gene flow among geographical groups was interrupted. A late Pleistocene demographic expansion was detected in the eastern geographical groups, while the expansion time (0.2-0.4 Mya) was earlier than the Last Glacial Maximum. CONCLUSION: It is proposed that the complicated topology preserves high genetic diversity and ancient lineages for geographical groups of A. morrisonia in China mainland and its two major islands, and restricts gene exchange during climate oscillations. Isolation by distance seems to be an important factor of genetic structure formation within geographical populations. Although glacial influence to population fluctuation was observed in late Pleistocene, it seems that populations in eastern China were more susceptible to climate change, and all geographical groups were growing stably through the Last Glacial Maximum. Coalescence analysis suggested that the ancestor of A. morrisonia might be traced back to the late Miocene, and the current phylogeographical structure of A. morrisonia is more likely to be attributable to a series geological events than to Pleistocene glacial cycles. PMID- 19558701 TI - Outbreaks of Streptococcus pneumoniae carriage in day care cohorts in Finland - implications for elimination of transmission. AB - BACKGROUND: Day care centre (DCC) attendees play a central role in maintaining the circulation of Streptococcus pneumoniae (pneumococcus) in the population. Exposure within families and within DCCs are the main risk factors for colonisation with pneumococcal serotypes in DCC attendees. METHODS: Transmission of serotype specific carriage was analysed with a continuous time event history model, based on longitudinal data from day care attendees and their family members. Rates of acquisition, conditional on exposure, were estimated in a Bayesian framework utilising latent processes of carriage. To ensure a correct level of exposure, non-participating day care attendees and their family members were included in the analysis. Posterior predictive simulations were used to quantify transmission patterns within day care cohorts, to estimate the basic reproduction number for pneumococcal carriage in a population of day care cohorts, and to assess the critical vaccine efficacy against carriage to eliminate pneumococcal transmission. RESULTS: The model, validated by posterior predictive sampling, was successful in capturing the strong temporal clustering of pneumococcal serotypes in the day care cohorts. In average 2.7 new outbreaks of pneumococcal carriage initiate in a day care cohort each month. While 39% of outbreaks were of size one, the mean outbreak size was 7.6 individuals and the mean length of an outbreak was 2.8 months. The role of families in creating and maintaining transmission was minimal, as only 10% of acquisitions in day care attendees were from family members. Considering a population of day care cohorts, a child-to-child basic reproduction number was estimated as 1.4 and the critical vaccine efficacy against acquisition of carriage as 0.3. CONCLUSION: Pneumococcal transmission occurs in serotype specific outbreaks of carriage, driven by within day-care transmission and between-serotype competition. An amplifying effect of the day care cohorts enhances the spread of pneumococcal serotypes within the population. The effect of vaccination, in addition to reducing susceptibility to pneumococcal carriage in the vaccinated, induces a herd effect, thus creating a counter-effect to the amplifying effect of the cohort. Consequently, the critical vaccine efficacy against carriage, required for elimination of transmission, is relatively low. Use of pneumococcal conjugate vaccines is expected to induce a notable herd protection against pneumococcal disease. PMID- 19558702 TI - Gender differences in the use of transportation services to community rehabilitation programs. AB - BACKGROUND: Prevention and reduction of disability among community-dwelling older adults have been an important health policy concern in Japan. Moreover, it has also become a gendered issue due to the recent rapid growth in older females than males with disability living in their own homes. The aim of this study is to examine whether there is a gender difference in the use of community rehabilitation programs in Japan, and if so, whether the lack of transportation services and accompanying caregivers are the reasons for the gender difference. METHODS: This study was based on surveys of the program administrators and the primary caregivers of the program participants from 55 randomly selected community rehabilitation programs (CRP) in the Tokyo metropolitan area. Questions included sociodemographic characteristics of program participants, types of transportation services provided by the CRP, caregiver's relationship to participant, and the nature of family support. Bivariate statistical analysis was conducted. RESULTS: Although there were more females than males with disability residing in communities, our findings showed that females were less likely to use CRP than males (1.3% and 2.3%, respectively; Chi2 = 93.0, p < 0.0001). Lower CRP use by females was related to lower availability of transportation services (36% without transportation service and 46% door-to-door services) and fewer caregivers accompanying the participants to CRP. CONCLUSION: This study builds on previous research findings, which suggest gender inequality in access to CRP. PMID- 19558703 TI - ResBoost: characterizing and predicting catalytic residues in enzymes. AB - BACKGROUND: Identifying the catalytic residues in enzymes can aid in understanding the molecular basis of an enzyme's function and has significant implications for designing new drugs, identifying genetic disorders, and engineering proteins with novel functions. Since experimentally determining catalytic sites is expensive, better computational methods for identifying catalytic residues are needed. RESULTS: We propose ResBoost, a new computational method to learn characteristics of catalytic residues. The method effectively selects and combines rules of thumb into a simple, easily interpretable logical expression that can be used for prediction. We formally define the rules of thumb that are often used to narrow the list of candidate residues, including residue evolutionary conservation, 3D clustering, solvent accessibility, and hydrophilicity. ResBoost builds on two methods from machine learning, the AdaBoost algorithm and Alternating Decision Trees, and provides precise control over the inherent trade-off between sensitivity and specificity. We evaluated ResBoost using cross-validation on a dataset of 100 enzymes from the hand-curated Catalytic Site Atlas (CSA). CONCLUSION: ResBoost achieved 85% sensitivity for a 9.8% false positive rate and 73% sensitivity for a 5.7% false positive rate. ResBoost reduces the number of false positives by up to 56% compared to the use of evolutionary conservation scoring alone. We also illustrate the ability of ResBoost to identify recently validated catalytic residues not listed in the CSA. PMID- 19558705 TI - Estimation of undernutrition and mean calorie intake in Africa: methodology, findings and implications. AB - BACKGROUND: As poverty and hunger are basic yardsticks of underdevelopment and destitution, the need for reliable statistics in this domain is self-evident. While the measurement of poverty through surveys is relatively well documented in the literature, for hunger, information is much scarcer, particularly for adults, and very different methodologies are applied for children and adults. Our paper seeks to improve on this practice in two ways. One is that we estimate the prevalence of undernutrition in sub-Saharan Africa (SSA) for both children and adults based on anthropometric data available at province or district level, and secondly, we estimate the mean calorie intake and implied calorie gap for SSA, also using anthropometric data on the same geographical aggregation level. RESULTS: Our main results are, first, that we find a much lower prevalence of hunger than presented in the Millennium Development reports (17.3% against 27.8% for the continent as a whole). Secondly, we find that there is much less spread in mean calorie intake across the continent than reported by the Food and Agricultural Organization (FAO) in the State of Food and Agriculture, 2007, the only estimate that covers the whole of Africa. While FAO estimates for calorie availability vary from a low of 1760 Kcal/capita/day for Central Africa to a high of 2825 Kcal/capita/day for Southern Africa, our estimates lay in a range of 2245 Kcal/capita/day (Eastern Africa) to 2618 Kcal/capita/day for Southern Africa. Thirdly, we validate the main data sources used (the Demographic and Health Surveys) by comparing them over time and with other available data sources for various countries. CONCLUSION: We conclude that the picture of Africa that emerges from anthropometric data is much less negative than that usually presented. Especially for Eastern and Central Africa, the nutritional status is less critical than commonly assumed and also mean calorie intake is higher, which implies that agricultural production and hence income must also have been growing at a pace at least high enough to keep up with population growth. In terms of methodology, our estimates form a base line for 2005 for the whole continent that can be easily updated with far less information for individual countries, as we show in an example for Ethiopia. PMID- 19558704 TI - Phylogenetic analysis and molecular evolution of the dormancy associated MADS-box genes from peach. AB - BACKGROUND: Dormancy associated MADS-box (DAM) genes are candidates for the regulation of growth cessation and terminal bud formation in peach. These genes are not expressed in the peach mutant evergrowing, which fails to cease growth and enter dormancy under dormancy-inducing conditions. We analyzed the phylogenetic relationships among and the rates and patterns of molecular evolution within DAM genes in the phylogenetic context of the MADS-box gene family. RESULTS: The peach DAM genes grouped with the SVP/StMADS11 lineage of type II MIKCC MADS-box genes. Phylogenetic analyses suggest that the peach SVP/StMADS11-like gene family, which contains significantly more members than annual model plants, expanded through serial tandem gene duplication. We found evidence of strong purifying selection acting to constrain functional divergence among the peach DAM genes and only a single codon, located in the C-terminal region, under significant positive selection. CONCLUSION: Because all DAM genes are expressed in peach and are subjected to strong purifying selection we suggest that the duplicated genes have been maintained by subfunctionalization and/or neofunctionalization. In addition, this pattern of selection suggests that the DAM genes are important for peach growth and development. PMID- 19558706 TI - A Bayesian approach to efficient differential allocation for resampling-based significance testing. AB - BACKGROUND: Large-scale statistical analyses have become hallmarks of post genomic era biological research due to advances in high-throughput assays and the integration of large biological databases. One accompanying issue is the simultaneous estimation of p-values for a large number of hypothesis tests. In many applications, a parametric assumption in the null distribution such as normality may be unreasonable, and resampling-based p-values are the preferred procedure for establishing statistical significance. Using resampling-based procedures for multiple testing is computationally intensive and typically requires large numbers of resamples. RESULTS: We present a new approach to more efficiently assign resamples (such as bootstrap samples or permutations) within a nonparametric multiple testing framework. We formulated a Bayesian-inspired approach to this problem, and devised an algorithm that adapts the assignment of resamples iteratively with negligible space and running time overhead. In two experimental studies, a breast cancer microarray dataset and a genome wide association study dataset for Parkinson's disease, we demonstrated that our differential allocation procedure is substantially more accurate compared to the traditional uniform resample allocation. CONCLUSION: Our experiments demonstrate that using a more sophisticated allocation strategy can improve our inference for hypothesis testing without a drastic increase in the amount of computation on randomized data. Moreover, we gain more improvement in efficiency when the number of tests is large. R code for our algorithm and the shortcut method are available at http://people.pcbi.upenn.edu/~lswang/pub/bmc2009/. PMID- 19558707 TI - Unexpected relationships of substructured populations in Chinese Locusta migratoria. AB - BACKGROUND: Highly migratory species are usually expected to have minimal population substructure because strong gene flow has the effect of homogenizing genetic variation over geographical populations, counteracting random drift, selection and mutation. The migratory locust Locusta migratoria belongs to a monotypic genus, and is an infamous pest insect with exceptional migratory ability - with dispersal documented over a thousand kilometers. Its distributional area is greater than that of any other locust or grasshopper, occurring in practically all the temperate and tropical regions of the eastern hemisphere. Consequently, minimal population substructuring is expected. However, in marked contrast to its high dispersal ability, three geographical subspecies have been distinguished in China, with more than nine being biologically and morphologically identified in the world. Such subspecies status has been under considerable debate. RESULTS: By multilocus microsatellite genotyping analysis, we provide ample genetic evidence for strong population substructure in this highly migratory insect that conforms to geography. More importantly, our genetic data identified an unexpected cryptic subdivision and demonstrated a strong affiliation of the East China locusts to those in Northwest/Northern China. The migratory locusts in China formed three distinct groups, viz. (1) the Tibetan group, comprising locusts from Tibet and nearby West China high mountain regions; this is congruent with the previously recognized Tibetan subspecies, L. m. tibetensis; (2) the South China group, containing locusts from the Hainan islands; this corresponds to the Southeast Asia oriental tropical subspecies L. m. manilensis; (3) the North China group, including locusts from the Northwest and Northern China (the Asiatic subspecies L. m. migratoria), Central China and Eastern China regions. Therefore, the traditional concept on Locusta subspecies status established from Uvarov in 1930s needs to be revised. The three groups of locusts probably have separate evolutionary histories that were most likely linked to Quaternary glaciations events, and derived from different ancestral refugial populations following postglacial expansions. CONCLUSION: The migratory locust populations in China have differentiated into three genetically distinct groups despite high dispersal capability. While this clarified long-standing suspicions on the subspecific diversification of this species in China, it also revealed that the locusts in the vast area of East China are not the oriental subspecies but the Asiatic subspecies, an unexpected substructuring pattern. The distribution pattern of the three locust groups in China may be primarily defined by adaptive differentiation coupled to Quaternary glaciations events. Our results are of general significance both for locust research and for phylogeographical study of flora and fauna in China, illustrating the potential importance of phylogeographical history in shaping the divergence and distribution patterns of widespread species with strong dispersal ability. PMID- 19558708 TI - Sub-typing of renal cell tumours; contribution of ancillary techniques. AB - BACKGROUND: Adult renal epithelial neoplasms are a heterogeneous group with varying prognosis and outcome requiring sub-classification. METHODS: Cases of renal cell carcinoma (RCC) in a 10 years period were analyzed with regard to the clinical features and histology. Sections were reviewed by four pathologists and the discordant cases were resolved with the help of Hale's colloidal iron stain, vimentin, CK 7, and vinculin immunostains and electron microscopy. RESULTS: Amongst the total of 278 cases, clear cell renal cell carcinoma was the commonest tumor with 74.8% cases, followed by papillary RCC 12.2%, chromophobe RCC 7.9%, oncocytoma 1.8%, and one case of collecting duct RCC. Eight cases were of sarcomatoid renal cell carcinoma. In 28/278 cases, diagnoses varied amongst the four pathologists and the discordance was resolved by Hale's colloidal iron stain, CK7 immunostain and electron microscopy. Vimentin and vinculin did not contribute much in differentiating subtypes of renal cell carcinomas. Relative incidence of sub-types of RCCs was compared with other series. CONCLUSION: To accurately subclassify renal cell carcinomas, simple ancillary techniques would possibly resolve all difficult cases. The relative incidence of sub-types of renal cell carcinoma is relatively consistent the world over. However, in India, RCCs afflict the patients two decades earlier. PMID- 19558710 TI - Adaptive radiation of gobies in the interstitial habitats of gravel beaches accompanied by body elongation and excessive vertebral segmentation. AB - BACKGROUND: The seacoasts of the Japanese Arc are fringed by many gravel beaches owing to active tectonic uplift and intense denudation caused by heavy rainfall. These gravel beaches are inhabited by gobies of the genus Luciogobius that burrow into the gravel sediment and live interstitially. Although their habitat and morphology (e. g., reduced fins, elongated, scale-less body, and highly segmented vertebral column) are highly unusual among fishes, little is known on how their morphological evolution has facilitated the colonization of interstitial habitats and promoted extensive diversification. We conducted thorough sampling of Luciogobius and related species throughout Japan, and performed molecular phylogenetic analysis to explore the patterns of morphological evolution associated with gravel beach colonization. RESULTS: An analysis of the mitochondrial cytochrome b gene suggested a remarkable diversity of previously unrecognized species. The species-level phylogeny based on six protein-coding nuclear genes clearly indicated that interstitial species cluster into two distinct clades, and that transitions from benthic or demersal habits to interstitial habits are strongly correlated with an increase in vertebral number. Colonization of gravel beach habitats is estimated to have occurred ca. 10 Ma, which coincides with the period of active orogenesis of the Japanese landmass. Different species of interstitial Luciogobius inhabit sediments with different granulometric properties, suggesting that microhabitat partitioning has been an important mechanism facilitating speciation in these fishes. CONCLUSION: This is the first study to document the adaptation to interstitial habitats by a vertebrate. Body elongation and excessive vertebral segmentation had been the key aspects enhancing body flexibility and fishes' ability to burrow into the gravel sediment. The rich diversity of coastal gravel habitats of the Japanese Arc has likely promoted the adaptive radiation of these unique gravel-dwelling fishes. PMID- 19558709 TI - Retinal pigment epithelial cells secrete neurotrophic factors and synthesize dopamine: possible contribution to therapeutic effects of RPE cell transplantation in Parkinson's disease. AB - BACKGROUND: New strategies for the treatment of Parkinson's disease (PD) are shifted from dopamine (DA) replacement to regeneration or restoration of the nigro-striatal system. A cell therapy using human retinal pigment epithelial (RPE) cells as substitution for degenerated dopaminergic (DAergic) neurons has been developed and showed promising prospect in clinical treatment of PD, but the exact mechanism underlying this therapy is not fully elucidated. In the present study, we investigated whether the beneficial effects of this therapy are related to the trophic properties of RPE cells and their ability to synthesize DA. METHODS: We evaluated the protective effects of conditioned medium (CM) from cultured RPE cells on the DAergic cells against 6-hydroxydopamine (6-OHDA)- and rotenone-induced neurotoxicity and determined the levels of glial cell derived neurotrophic factor (GDNF) and brain derived neurotrophic factor (BDNF) released by RPE cells. We also measured the DA synthesis and release. Finally we transplanted microcarriers-RPE cells into 6-OHDA lesioned rats and observed the improvement in apomorphine-induced rotations (AIR). RESULTS: We report here: (1) CM from RPE cells can secret trophic factors GDNF and BDNF, and protect DAergic neurons against the 6-OHDA- and rotenone-induced cell injury; (2) cultured RPE cells express L-dopa decarboxylase (DDC) and synthesize DA; (3) RPE cells attached to microcarriers can survive in the host striatum and improve the AIR in 6-OHDA-lesioned animal model of PD; (4) GDNF and BDNF levels are found significantly higher in the RPE cell-grafted tissues. CONCLUSION: These findings indicate the RPE cells have the ability to secret GDNF and BDNF, and synthesize DA, which probably contribute to the therapeutic effects of RPE cell transplantation in PD. PMID- 19558711 TI - Truncation in the tcdC region of the Clostridium difficile PathLoc of clinical isolates does not predict increased biological activity of Toxin B or Toxin A. AB - BACKGROUND: The increased severity of disease associated with the NAP1 strain of Clostridium difficile has been attributed to mutations to the tcdC gene which codes for a negative regulator of toxin production. To assess the role of hyper production of Toxins A and B in clinical isolates of Clostridium difficile, two NAP1-related and five NAP1 non-related strains were compared. METHODS: Sequencing was performed on tcdC, tcdR, and tcdE to determine if there were differences that might account for hyper-production of Toxin A and Toxin B in NAP1-related strains. Biological activity of Toxin B was evaluated using the HFF cell CPE assay and Toxin A biological activity was assessed using the Caco-2 Trans membrane resistance assay. RESULTS: Our results confirm that Toxin A and Toxin B production in NAP1-related strains and ATCC 43255 occurs earlier in the exponential growth phase compared to most NAP1-nonrelated clinical isolates. Despite the hyper-production observed in ATCC 43255 it had no mutations in tcdC, tcdR or tcdE. Analysis of the other clinical isolates indicated that the kinetics and ultimate final concentration of Toxin A and B did not correlate with the presence or lack of alterations in tcdC, tcdR or tcdE. CONCLUSION: Our data do not support a direct role for alterations in the tcdC gene as a predictor of hyperproduction of Toxin A and B in NAP1-related strains. PMID- 19558712 TI - Evaluation of community level interventions to address social and structural determinants of health: a cluster randomised controlled trial. AB - BACKGROUND: In London and the rest of the UK, diseases associated with poor diet, inadequate physical activity and mental illness account for a large proportion of area based health inequality. There is a lack of evidence on interventions promoting healthier behaviours especially in marginalised populations, at a structural or ecological level and utilising a community development approach.The Well London project financed by the Big Lottery 'Wellbeing' Fund and implemented by a consortium of London based agencies led by the Greater London Authority and the London Health Commission is implementing a set of complex interventions across 20 deprived areas of London. The interventions focus on healthy eating, healthy physical activity and mental health and wellbeing and are designed and executed with community participation complementing existing facilities and services. METHODS/DESIGN: The programme will be evaluated through a cluster randomised controlled trial. Forty areas across London were chosen based on deprivation scores. Areas were characterised by high proportion of Black and Minority Ethnic residents, worklessness, ill-health and poor physical environments. Twenty areas were randomly assigned to the intervention arm of Well London project and twenty 'matched' areas assigned as controls. Measures of physical activity, diet and mental health are collected at start and end of the project and compared to assess impact.The quantitative element will be complemented by a longitudinal qualitative study elucidating pathways of influence between intervention activities and health outcomes. A related element of the study investigates the health-related aspects of the structural and ecological characteristics of the project areas. The project 'process' will also be evaluated. DISCUSSION: The size of the project and the fact that the interventions are 'complex' in the sense that firstly, there are a number of interacting components with a wide range of groups and organisational levels targeted by the intervention, and secondly, a degree of flexibility or tailoring of the intervention, makes this trial potentially very useful in providing evidence of the types of activities that can be used to address chronic health problems in communities suffering from multiple deprivation. TRIAL REGISTRATION: Current Controlled Trials ISRCTN68175121. PMID- 19558714 TI - Ethnic differences in cancer incidence in Estonia: two cross-sectional unlinked census-based cancer incidence analyses. AB - BACKGROUND: Estonian and Russian ethnic groups in Estonia differ from one another in several aspects, such as historic and socio-economic background, language and culture. The aim of the current study was to examine ethnic differences in cancer incidence in Estonia, and to compare the situation before and after the profound political and economical changes in the early 1990s. METHODS: Two cross-sectional unlinked census-based cancer incidence analyses were performed. Cancer incidence data were obtained from the Estonian Cancer Registry. Population denominators came from the population censuses of 1989 and 2000. Standardized cancer incidence rates were calculated for men and women for the aggregate periods 1988-1990 and 1999-2000. Differences in cancer incidence between Estonians and Russians in 1989 and 2000 were estimated for both sexes, using standardized rate ratios with 95% confidence intervals. RESULTS: In 1988-1990, the total cancer incidence in Russian men was higher than that in Estonian men (SRR = 1.26, 95%CI = 1.19-1.34). In 1999-2000, the total cancer incidence in men showed only slightly higher estimates in Russians than in Estonians (SRR = 1.06, 95%CI = 0.99-1.32). Cancers of stomach, colon and lung had persisting higher values in Russian men in 1999 2000. In women, the differences were smaller than in men, and the total cancer incidence showed no differences relating to neither of the time periods studied. With regard to specific sites, excess of stomach cancer incidence was seen in Russian women (SRR = 1.45, 95%CI = 1.15-1.81). The ethnic differences in general decreased between the two time periods studied. CONCLUSION: Some of the differences in cancer rates between the Estonians and Russians in Estonia are likely to be attributable to the variation in exposure to specific etiologic factors that are causedby differences in lifestyle and habits, such as hygiene, smoking and drinking. Further research with a view to understanding these ethnic differences in cancer incidence is warranted. PMID- 19558713 TI - Role of the H1 haplotype of microtubule-associated protein tau (MAPT) gene in Greek patients with Parkinson's disease. AB - BACKGROUND: The extended tau haplotype (H1) that covers the entire human microtubule-associated protein tau (MAPT) gene has been implicated in Parkinson's disease (PD). Nevertheless, controversial results, such as two studies in Greek populations with opposite effects, have been reported. Therefore, we set out to determine whether the H1 haplotype and additional single nucleotide polymorphisms (SNPs) included in H1 are associated with PD in a sample of Greek patients. METHODS: We analysed MAPT haplotypes in cohorts of 122 patients and 123 controls of Greek origin, respectively. SNP genotyping was performed with Taqman assays and genotyping results were confirmed by sequencing. RESULTS: The presence of the H1 haplotype was significantly associated with PD (odds ratio for H1H1 vs. H1H2 and H2H2: 1.566; 95% CI: 1.137-2.157; P = 0.006) and remained so after adjustment for sex. Further analysis of H1 sub-haplotypes with three single nucleotide polymorphisms (rs242562, rs2435207 and rs3785883) demonstrated no significant association with PD. CONCLUSION: Our data support the overall genetic role of MAPT and the H1 haplotype for PD susceptibility in Greek patients. However, the previously supported association of H1 sub-haplotypes with PD could not be confirmed in our study. PMID- 19558716 TI - Clinicians' evaluations of, endorsements of, and intentions to use practice guidelines change over time: a retrospective analysis from an organized guideline program. AB - PURPOSE: Clinical practice guidelines (CPGs) can improve clinical care but uptake and application are inconsistent. Objectives were: to examine temporal trends in clinicians' evaluations of, endorsements of, and intentions to use cancer CPGs developed by an established CPG program; and to evaluate how predictor variables (clinician characteristics, beliefs, and attitudes) are associated with these trends. DESIGN AND METHODS: Between 1999 and 2005, 756 clinicians evaluated 84 Cancer Care Ontario CPGs, yielding 4,091 surveys that targeted four CPG quality domains (rigour, applicability, acceptability, and comparative value), clinicians' endorsement levels, and clinicians' intentions to use CPGs in practice. RESULTS: Time: In contrast to the applicability and intention to use in practice scores, there were small but statistically significant annual net gains in ratings for rigour, acceptability, comparative value, and CPG endorsement measures (p < 0.05 for all rating categories). PREDICTORS: In 17 comparisons, ratings were significantly higher among clinicians having the most favourable beliefs and most positive attitudes and lowest for those having the least favourable beliefs and most negative attitudes (p < 0.05). Interactions Time x PREDICTORS: Over time, differences in outcomes among clinicians decreased due to positive net gains in scores by clinicians whose beliefs and attitudes were least favorable. CONCLUSION: Individual differences among clinicians largely explain variances in outcomes measured. Continued engagement of clinicians least receptive to CPGs may be worthwhile because they are the ones showing most significant gains in CPG quality ratings, endorsement ratings, and intentions to use in practice ratings. PMID- 19558715 TI - Modelling dominance in a flexible intercross analysis. AB - BACKGROUND: The aim of this paper is to develop a flexible model for analysis of quantitative trait loci (QTL) in outbred line crosses, which includes both additive and dominance effects. Our flexible intercross analysis (FIA) model accounts for QTL that are not fixed within founder lines and is based on the variance component framework. Genome scans with FIA are performed using a score statistic, which does not require variance component estimation. RESULTS: Simulations of a pedigree with 800 F2 individuals showed that the power of FIA including both additive and dominance effects was almost 50% for a QTL with equal allele frequencies in both lines with complete dominance and a moderate effect, whereas the power of a traditional regression model was equal to the chosen significance value of 5%. The power of FIA without dominance effects included in the model was close to those obtained for FIA with dominance for all simulated cases except for QTL with overdominant effects. A genome-wide linkage analysis of experimental data from an F2 intercross between Red Jungle Fowl and White Leghorn was performed with both additive and dominance effects included in FIA. The score values for chicken body weight at 200 days of age were similar to those obtained in FIA analysis without dominance. CONCLUSION: We have extended FIA to include QTL dominance effects. The power of FIA was superior, or similar, to standard regression methods for QTL effects with dominance. The difference in power for FIA with or without dominance is expected to be small as long as the QTL effects are not overdominant. We suggest that FIA with only additive effects should be the standard model to be used, especially since it is more computationally efficient. PMID- 19558717 TI - Spatial analysis of plague in California: niche modeling predictions of the current distribution and potential response to climate change. AB - BACKGROUND: Plague, caused by the bacterium Yersinia pestis, is a public and wildlife health concern in California and the western United States. This study explores the spatial characteristics of positive plague samples in California and tests Maxent, a machine-learning method that can be used to develop niche-based models from presence-only data, for mapping the potential distribution of plague foci. Maxent models were constructed using geocoded seroprevalence data from surveillance of California ground squirrels (Spermophilus beecheyi) as case points and Worldclim bioclimatic data as predictor variables, and compared and validated using area under the receiver operating curve (AUC) statistics. Additionally, model results were compared to locations of positive and negative coyote (Canis latrans) samples, in order to determine the correlation between Maxent model predictions and areas of plague risk as determined via wild carnivore surveillance. RESULTS: Models of plague activity in California ground squirrels, based on recent climate conditions, accurately identified case locations (AUC of 0.913 to 0.948) and were significantly correlated with coyote samples. The final models were used to identify potential plague risk areas based on an ensemble of six future climate scenarios. These models suggest that by 2050, climate conditions may reduce plague risk in the southern parts of California and increase risk along the northern coast and Sierras. CONCLUSION: Because different modeling approaches can yield substantially different results, care should be taken when interpreting future model predictions. Nonetheless, niche modeling can be a useful tool for exploring and mapping the potential response of plague activity to climate change. The final models in this study were used to identify potential plague risk areas based on an ensemble of six future climate scenarios, which can help public managers decide where to allocate surveillance resources. In addition, Maxent model results were significantly correlated with coyote samples, indicating that carnivore surveillance programs will continue to be important for tracking the response of plague to future climate conditions. PMID- 19558718 TI - Cell-free protein synthesis energized by slowly-metabolized maltodextrin. AB - BACKGROUND: Cell-free protein synthesis (CFPS) is a rapid and high throughput technology for obtaining proteins from their genes. The primary energy source ATP is regenerated from the secondary energy source through substrate phosphorylation in CFPS. RESULTS: Distinct from common secondary energy sources (e.g., phosphoenolpyruvate - PEP, glucose-6-phosphate), maltodextrin was used for energizing CFPS through substrate phosphorylation and the glycolytic pathway because (i) maltodextrin can be slowly catabolized by maltodextrin phosphorylase for continuous ATP regeneration, (ii) maltodextrin phosphorylation can recycle one phosphate per reaction for glucose-1-phosphate generation, and (iii) the maltodextrin chain-shortening reaction can produce one ATP per glucose equivalent more than glucose can. Three model proteins, esterase 2 from Alicyclobacillus acidocaldarius, green fluorescent protein, and xylose reductase from Neurospora crassa were synthesized for demonstration. CONCLUSION: Slowly-metabolized maltodextrin as a low-cost secondary energy compound for CFPS produced higher levels of proteins than PEP, glucose, and glucose-6-phospahte. The enhancement of protein synthesis was largely attributed to better-controlled phosphate levels (recycling of inorganic phosphate) and a more homeostatic reaction environment. PMID- 19558719 TI - The UK Quality and Outcomes Framework pay-for-performance scheme and spirometry: rewarding quality or just quantity? A cross-sectional study in Rotherham, UK. AB - BACKGROUND: Accurate spirometry is important in the management of COPD. The UK Quality and Outcomes Framework pay-for-performance scheme for general practitioners includes spirometry related indicators within its COPD domain. It is not known whether high achievement against QOF spirometry indicators is associated with spirometry to BTS standards. METHODS: Data were obtained from the records of 3,217 patients randomly sampled from 5,649 patients with COPD in 38 general practices in Rotherham, UK. Severity of airflow obstruction was categorised by FEV1 (% predicted) according to NICE guidelines. This was compared with clinician recorded COPD severity. The proportion of patients whose spirometry met BTS standards was calculated in each practice using a random sub sample of 761 patients. The Spearman rank correlation between practice level QOF spirometry achievement and performance against BTS spirometry standards was calculated. RESULTS: Spirometry as assessed by clinical records was to BTS standards in 31% of cases (range at practice level 0% to 74%). The categorisation of airflow obstruction according to the most recent spirometry results did not agree well with the clinical categorisation of COPD recorded in the notes (Cohen's kappa = 0.34, 0.30 - 0.38). 12% of patients on COPD registers had FEV1 (% predicted) results recorded that did not support the diagnosis of COPD. There was no association between quality, as measured by adherence to BTS spirometry standards, and either QOF COPD9 achievement (Spearman's rho = -0.11), or QOF COPD10 achievement (rho = 0.01). CONCLUSION: The UK Quality and Outcomes Framework currently assesses the quantity, but not the quality of spirometry. PMID- 19558720 TI - Early MRI response monitoring of patients with advanced hepatocellular carcinoma under treatment with the multikinase inhibitor sorafenib. AB - BACKGROUND: New therapeutic principles in clinical oncology require the adjustment of response criteria to govern therapy decisions. For advanced hepatocellular carcinoma (HCC) a new era has recently begun by the approval of the multikinase inhibitor sorafenib. As a unique feature, HCC usually develops in a diseased liver and current imaging technologies employing classical response criteria have not been prospectively evaluated for this new treatment. METHODS: MRI signal patterns were assessed in 21 advanced HCC patients receiving sorafenib. MRI was performed at baseline and in short-term intervals thereafter. Signal changes under therapy on T1WI, T2WI and post-gadolinium images including necrosis volume and its ratio to the entire tumor volume were compared to baseline imaging. To assess the association between the categorical variables, Fisher's exact tests were applied for a statistical analysis. Survey time ranged from 2-65 weeks, and a total of 39 target lesions were evaluated. RESULTS: Signal abnormalities during sorafenib therapy were disclosed by T1WI and T2WI in 15/21 patients. The predominant tumor signal change was hyperintensity on both T1WI and T2WI. Interestingly, most patients developed MRI signal changes within 4 weeks of therapy; in contrast, two non-responders did not show any signal alteration at follow-up. Under therapy, 16/21 patients presented with new or progressive necrosis, whereas 7 patients achieved temporarily >75% tumor necrosis under sorafenib. Significantly associated MRI variables were increase in T1WI signal and tumor necrosis (p = 0.017) as well as increase of tumor necrosis with an elevated ratio of necrotic to vital tumor areas (p = 0.002). Remarkably, some (3/13) of the patients developing necrotic tumor areas showed a relevant (>20%) increase in tumor volume, which should be considered in the assessment of imaging studies. CONCLUSION: As sorafenib induces early intralesional necrosis with profound changes in T1WI/T2WI MRI signal intensities and measurable necrotic tumor areas in most HCC patients, early MRI-based evaluation could pave the way for its rationale and cost-effective application. PMID- 19558721 TI - Transplantation of selected or transgenic blood stem cells - a future treatment for HIV/AIDS? AB - Interaction with the chemokine receptor, CCR5, is a necessary precondition for maintaining HIV-1 infection. Individuals with the CCR5-delta32 deletion who lack this receptor are highly resistant to infection by the most common forms of HIV 1. We recently reported on the successful transplantation in an HIV-1-positive patient of allogeneic stem cells homozygous for the CCR5-delta32 allele, which stopped viral replication for more than 27 months without antiretroviral therapy.Here, we report on the results of a meeting regarding the potential implications and future directions of stem cell-targeted HIV treatments. The meeting drew together an international panel of hematologists, immunologists, HIV specialists and representatives from bone marrow donor registries.The meeting came to an agreement to support further attempts to use CCR5-delta32 deleted stem cells, for example, prescreened cord blood stem cells, to treat probable HIV-1 positive patients with malignancies. Furthermore, improvement of HIV-1 therapy that interferes with the entry mechanism seems to be a promising approach in HIV 1-infected patients with no matching CCR5-delta32 deleted donor. PMID- 19558722 TI - The efficacy of iron chelator regimes in reducing cardiac and hepatic iron in patients with thalassaemia major: a clinical observational study. AB - BACKGROUND: Available iron chelation regimes in thalassaemia may achieve different changes in cardiac and hepatic iron as assessed by MR. The aim of this study was to assess the efficacy of four available iron chelator regimes in 232 thalassaemia major patients by assessing the rate of change in repeated measurements of cardiac and hepatic MR. RESULTS: For the heart, deferiprone and the combination of deferiprone and deferoxamine significantly reduced cardiac iron at all levels of iron loading. As patients were on deferasirox for a shorter time, a second analysis ("Initial interval analysis") assessing the change between the first two recorded MR results for both cardiac and hepatic iron (minimum interval 12 months) was made. Combination therapy achieved the most rapid fall in cardiac iron load at all levels and deferiprone alone was significantly effective with moderate and mild iron load. In the liver, deferasirox effected significant falls in iron load and combination therapy resulted in the most rapid decline. CONCLUSION: With the knowledge of the efficacy of the different available regimes and the specific iron load in the heart and the liver, appropriate tailoring of chelation therapy should allow clearance of iron. Combination therapy is best in reducing both cardiac and hepatic iron, while monotherapy with deferiprone or deferasirox are effective in the heart and liver respectively. The outcomes of this study may be useful to physicians as to the chelation they should prescribe according to the levels of iron load found in the heart and liver by MR. PMID- 19558723 TI - Comparative transcriptomics in the Triticeae. AB - BACKGROUND: Barley and particularly wheat are two grass species of immense agricultural importance. In spite of polyploidization events within the latter, studies have shown that genotypically and phenotypically these species are very closely related and, indeed, fertile hybrids can be created by interbreeding. The advent of two genome-scale Affymetrix GeneChips now allows studies of the comparison of their transcriptomes. RESULTS: We have used the Wheat GeneChip to create a "gene expression atlas" for the wheat transcriptome (cv. Chinese Spring). For this, we chose mRNA from a range of tissues and developmental stages closely mirroring a comparable study carried out for barley (cv. Morex) using the Barley1 GeneChip. This, together with large-scale clustering of the probesets from the two GeneChips into "homologous groups", has allowed us to perform a genomic-scale comparative study of expression patterns in these two species. We explore the influence of the polyploidy of wheat on the results obtained with the Wheat GeneChip and quantify the correlation between conservation in gene sequence and gene expression in wheat and barley. In addition, we show how the conservation of expression patterns can be used to elucidate, probeset by probeset, the reliability of the Wheat GeneChip. CONCLUSION: While there are many differences in expression on the level of individual genes and tissues, we demonstrate that the wheat and barley transcriptomes appear highly correlated. This finding is significant not only because given small evolutionary distance between the two species it is widely expected, but also because it demonstrates that it is possible to use the two GeneChips for comparative studies. This is the case even though their probeset composition reflects rather different design principles as well as, of course, the present incomplete knowledge of the gene content of the two species. We also show that, in general, the Wheat GeneChip is not able to distinguish contributions from individual homoeologs. Furthermore, the comparison between the two species leads us to conclude that the conservation of both gene sequence as well as gene expression is positively correlated with absolute expression levels, presumably reflecting increased selection pressure on genes coding for proteins present at high levels. In addition, the results indicate the presence of a correlation between sequence and expression conservation within the Triticeae. PMID- 19558724 TI - The ACTIVE cognitive training trial and predicted medical expenditures. AB - BACKGROUND: Health care expenditures for older adults are disproportionately high and increasing at both the individual and population levels. We evaluated the effects of the three cognitive training interventions (memory, reasoning, or speed of processing) in the ACTIVE study on changes in predicted medical care expenditures. METHODS: ACTIVE was a multisite randomized controlled trial of older adults (>or= 65). Five-year follow-up data were available for 1,804 of the 2,802 participants. Propensity score weighting was used to adjust for potential attrition bias. Changes in predicted annualmedical expenditures were calculated at the first and fifth annual follow-up assessments using a new method for translating functional status scores. Multiple linear regression methods were used in this cost-offset analysis. RESULTS: At one and five years post-training, annual predicted expenditures declinedby $223 (p = .024) and $128 (p = .309), respectively, in the speed of processing treatment group, but there were no statistically significant changes in the memory or reasoning treatment groups compared to the no-contact control group at either period. Statistical adjustment for age, race, education, MMSE scores, ADL and IADL performance scores, EPT scores, chronic condition counts, and the SF-36 PCS and MCS scores at baseline did not alter the one-year ($244; p = .012) or five-year ($143; p = .250) expenditure declines in the speed of processing treatment group. CONCLUSION: The speed of processing intervention significantly reduced subsequent annual predicted medical care expenditures at the one-year post-baseline comparison, but annual savings were no longer statistically significant at the five-year post baseline comparison. PMID- 19558726 TI - Effect of Fuzheng Huayu formula and its actions against liver fibrosis. AB - Liver fibrosis is a common histological process to develop into cirrhosis in various chronic liver diseases including chronic hepatitis and fatty liver. Therefore anti-liver fibrosis is very important strategy to treat chronic liver diseases. Fuzheng Huayu (FZHY), a preparation containing herbs such as Radix Salvia Miltiorrhizae, Cordyceps, Semen Persicae, was formulated on the basis of Chinese medicine theory in treating liver fibrosis and was approved. Pharmacological studies and clinical trials demonstrate that FZHY has a significant effect against liver fibrosis and that many of the pharmacological actions are attributable to the effect. This article reviews the effects and actions of FZHY, in particular the effects observed from clinical trials in treating liver fibrosis caused by chronic hepatitis B and the actions on inhibition of hepatic stellate cell activation, protection of hepatocytes and inhibition of hepatic sinusoidal capillarization. This article also reviews the coordinated effects of the constituent herbs of FZHY and the actions of their active compounds such as salvianonic acid B (SA-B) on liver fibrosis. PMID- 19558727 TI - Cerebellar mass as a primary presentation of papillary thyroid carcinoma: case report and literature review. AB - BACKGROUND: Papillary carcinoma is the most common differentiated malignant thyroid neoplasm. The biological course of this cancer is typically indolent with a protracted clinical course. Metastases commonly occur in regional lymph nodes, and distant metastasis is a late and rare occurrence. We report a patient who presented with cerebellar metastasis prior to the diagnosis of papillary thyroid carcinoma and review the literature of brain metastasis from papillary thyroid carcinoma. RESULTS: A 75-year old female presented at the emergency room with progressive dizziness, headache and vomiting, where a brain CT and MRI showed a posterior cerebellar tumor. Surgical resection revealed papillary carcinoma consistent with thyroid origin. Subsequent ultrasound and CT-scan revealed a thyroid nodule, after which the patient underwent total thyroidectomy. Pathologic evaluation was consistent with papillary thyroid carcinoma. CONCLUSION: Brain metastasis may rarely be the initial presentation of papillary thyroid carcinoma. Solitary brain metastasis can completely be resected with better prognosis. PMID- 19558725 TI - Expressed sequence tags from larval gut of the European corn borer (Ostrinia nubilalis): exploring candidate genes potentially involved in Bacillus thuringiensis toxicity and resistance. AB - BACKGROUND: Lepidoptera represents more than 160,000 insect species which include some of the most devastating pests of crops, forests, and stored products. However, the genomic information on lepidopteran insects is very limited. Only a few studies have focused on developing expressed sequence tag (EST) libraries from the guts of lepidopteran larvae. Knowledge of the genes that are expressed in the insect gut are crucial for understanding basic physiology of food digestion, their interactions with Bacillus thuringiensis (Bt) toxins, and for discovering new targets for novel toxins for use in pest management. This study analyzed the ESTs generated from the larval gut of the European corn borer (ECB, Ostrinia nubilalis), one of the most destructive pests of corn in North America and the western world. Our goals were to establish an ECB larval gut-specific EST database as a genomic resource for future research and to explore candidate genes potentially involved in insect-Bt interactions and Bt resistance in ECB. RESULTS: We constructed two cDNA libraries from the guts of the fifth-instar larvae of ECB and sequenced a total of 15,000 ESTs from these libraries. A total of 12,519 ESTs (83.4%) appeared to be high quality with an average length of 656 bp. These ESTs represented 2,895 unique sequences, including 1,738 singletons and 1,157 contigs. Among the unique sequences, 62.7% encoded putative proteins that shared significant sequence similarities (E-value or = 20%. All the patients then underwent a period of ventilation on the same settings: tidal volume 6 mL/kg, PEEP at the level set by the physician before the experiment, plateau pressure < 30 cm H(2)O. Then each patient underwent 3 ventilation strategies, each applied for one hour: optimal PEEP alone; optimal PEEP plus one sustained inflation (40 cm H(2)O for 30 s); and optimal PEEP plus sigh breaths (ie, twice the baseline tidal volume, plateau pressure < 40 cm H(2)O) every 25 breaths. After the application of each PEEP strategy we measured arterial blood gas values and the static compliance of the respiratory system. RESULTS: The mean +/- SD optimal PEEP was 12 +/- 4 cm H(2)O. The measurements from the standardization periods were comparable between the 3 PEEP groups. In the optimal-PEEP-plus-sighs group the changes in P(aO(2)) (85 +/- 96%) and static compliance (14 +/- 20%) were significantly greater than in the 2 other groups. CONCLUSIONS: Sighs superimposed on lung-protective mechanical ventilation with optimal PEEP improved oxygenation and static compliance in patients with early ALI/ARDS. PMID- 19558736 TI - Metered-dose inhaler with spacer instead of nebulizer during the outbreak of severe acute respiratory syndrome in Singapore. AB - BACKGROUND: The use of metered-dose inhaler (MDI) with spacer instead of nebulizer may be important during an outbreak of an airborne infection. However, there is a paucity of data on patients' and nurses' abilities and perspectives on MDI with spacer for the treatment of acute airway obstruction during such an outbreak. METHODS: We evaluated 50 consecutive MDI-with-spacer treatments administered in the respiratory wards of the National University Hospital of Singapore, and interviewed the patients after each treatment during the outbreak of severe acute respiratory syndrome (SARS). We also conducted interviews with 50 nurses who had experience in administering bronchodilators via both nebulizer and MDI with spacer. RESULTS: Forty-six patients (92%) were able to use MDI with spacer effectively. Sixteen percent of the patients preferred nebulizer over MDI with spacer. Fifty-eight percent of the patients thought MDI with spacer was easier to use than nebulizer, and 34% thought MDI was as easy to use as nebulizer. Sixteen percent of the patients thought that nebulizer was more effective than MDI with spacer in relieving their symptoms. Ninety-six percent of the nurses preferred nebulizer over MDI with spacer. Forty-two nurses (84%) thought that nebulizer was more effective for treating acute airflow obstruction in the hospital. CONCLUSIONS: In the in-patient setting during an outbreak of an airborne infection, for treatment of acute airflow obstruction, MDI with spacer was acceptable and preferred by a high percentage of patients. However, a high percentage of nurses had misconceptions regarding the efficacy of and patients' ability to use MDI with spacer. PMID- 19558737 TI - Improved outcomes with routine respiratory therapist evaluation of non-intensive care-unit surgery patients. AB - BACKGROUND: Respiratory therapist (RT) driven protocols decrease ventilator days and resource utilization in the intensive care unit (ICU). Protocols have been studied in non-ICU settings, but their effect on mortality has been incompletely studied. METHODS: In our neurosurgery step-down, trauma/surgery step-down, and trauma/surgery general units we initiated an RT-driven evaluate-and-treat protocol that included a standardized, quantitative, RT-driven patient-assessment scale and protocolized interventions. Before and after initiation of the protocol we collected data on non-ICU patients at risk for pulmonary complications. RESULTS: The patient groups before (n = 2,230) and after (n = 2,805) protocol initiation were well matched in age, sex, Charlson score, and admitting service. Most of the patients, whether assessed by a physician or an RT, were deemed to have low risk of pulmonary complications and did not require any respiratory treatments. The number of respiratory treatments increased after protocol initiation, but the patients who received respiratory treatments after protocol initiation had shorter ICU stay and hospital stay, and lower total hospital costs than those who received respiratory treatments before protocol initiation. There was a nonsignificant trend toward lower mortality after protocol initiation. CONCLUSIONS: Our RT-evaluate-and-treat protocol for non-ICU surgery patients was associated with more patients receiving respiratory treatments but decreased ICU and hospital stay and lower total hospital costs. Routine RT-driven assessment of non-ICU patients may reduce pulmonary complications in high-risk patients. PMID- 19558738 TI - Respiratory care clinical education: a needs assessment for preceptor training. AB - BACKGROUND: There has been a growing interest in the use of volunteer clinical preceptors to provide clinical instruction to respiratory therapy (RT) students. However, many RT preceptors have had little or no training in preceptorship. We sought to identify the preceptor training needs of programs that lead to the Registered Respiratory Therapist or Certified Respiratory Therapist credential (RT programs). METHODS: Via e-mail we asked the directors of accredited RT programs to respond to a Web-based survey. RESULTS: Seventy-four RT program directors, from programs across the United States, responded. Eighty-two percent of the respondents' programs offer an associate's degree and 16% offer a baccalaureate degree. The majority of the respondents' programs use unpaid clinical preceptors. Thirty-two percent of the respondents indicated that the preceptors had received no preceptor training. Among the preceptors who did receive training, the duration of training ranged from 1 hour to 6 weeks. The training was typically delivered by the director of clinical education or program faculty. Eighty-one percent of the respondents believed there is a need for a standardized preceptor-training program. The respondents' understanding of, curriculum for, and implementation of preceptor training differed considerably, and there were substantial differences in the content and duration of the existing preceptor-training programs. Seventy-two percent of the respondents had experienced barriers to preceptor training. CONCLUSIONS: A standardized preceptor training program is needed to improve the quality of preceptorship and assure that RT programs prepare graduates for 21st-century RT practice. PMID- 19558739 TI - Adult asthma disease management: an analysis of studies, approaches, outcomes, and methods. AB - BACKGROUND: Disease management has been implemented for patients with asthma in various ways. We describe the approaches to and components of adult asthma disease-management interventions, examine the outcomes evaluated, and assess the quality of published studies. METHODS: We searched the MEDLINE, EMBASE, CINAHL, PsychInfo, and Cochrane databases for studies published in 1986 through 2008, on adult asthma management. With the studies that met our inclusion criteria, we examined the clinical, process, medication, economic, and patient-reported outcomes reported, and the study designs, provider collaboration during the studies, and statistical methods. RESULTS: Twenty-nine articles describing 27 studies satisfied our inclusion criteria. There was great variation in the content, extent of collaboration between physician and non-physician providers responsible for intervention delivery, and outcomes examined across the 27 studies. Because of limitations in the design of 22 of the 27 studies, the differences in outcomes assessed, and the lack of rigorous statistical adjustment, we could not draw definitive conclusions about the effectiveness or cost-effectiveness of the asthma disease-management programs or which approach was most effective. CONCLUSIONS: Few well-designed studies with rigorous evaluations have been conducted to evaluate disease-management interventions for adults with asthma. Current evidence is insufficient to recommend any particular intervention. PMID- 19558740 TI - The physiology of dinosaurs: circulatory and respiratory function in the largest animals ever to walk the earth. AB - The cardiopulmonary physiology of dinosaurs-and especially of the long-necked sauropods, which grew much larger than any land animals before or since-should be inherently fascinating to anyone involved in respiratory care. What would the blood pressure be in an animal 12 m (40 ft) tall? How could airway resistance and dead space be overcome while breathing through a trachea 9 m (30 ft) long? The last decade has seen a dramatic increase in evidence bearing on these questions. Insight has come not only from new fossil discoveries but also from comparative studies of living species, clarification of evolutionary relationships, new evaluation techniques, computer modeling, and discoveries about the earth's ancient atmosphere. Pumping a vertical column of blood 8 m (26 ft) above the heart would probably require an arterial blood pressure > 600 mm Hg, and the implications of this for cardiac size and function have led to the proposal of several alternative cardiopulmonary designs. Diverse lines of evidence suggest that the giant sauropods were probably warm-blooded and metabolically active when young, but slowed their metabolism as they approached adult size, which diminished the load on the circulatory system. Circulatory considerations leave little doubt that the dinosaurs had 4-chambered hearts. Birds evolved from dinosaurs, and the avian-type air-sac respiratory system, which is more efficient than its mammalian counterpart, may hold the answer to the breathing problems posed by the sauropods' very long necks. Geochemical and other data indicate that, at the time the dinosaurs first appeared, the atmospheric oxygen concentration was only about half of what it is today, and development of the avian-type respiratory system may have been key in the dinosaurs' evolutionary success, enabling them to out-compete the mammals and dominate the land for 150 million years. PMID- 19558741 TI - What is the legacy of the National Institutes of Health Acute Respiratory Distress Syndrome Network? AB - It has been almost 15 years since the National Institutes of Health created the Acute Respiratory Distress Syndrome Clinical Trials Network (ARDS Network) and nearly a decade since the completion of the landmark low-tidal volume (V(T)) trial. In retrospect, the ARDS Network had a profound impact on the design and conduct of clinical trials in critical care. It represented the first time the federal government funded a clinical trials network devoted to Phase-III testing of important, non-pharmacologic therapies. Also the ARDS Network introduced factorial design into critical-care research, which allowed Phase-II testing of promising therapies. Other important contributions from the ARDS Network may not become apparent for many years. These include the ongoing mentoring of a new generation of critical-care researchers, as well as continued testing on an enormous store of biological samples that inevitably will advance our understanding of the pathogenesis of ARDS. Perhaps someday this may lead to another therapeutic breakthrough. Part of the ARDS Network's legacy surely will have been the opening of a dialog regarding the design of clinical trials in critical care, as well as a concerted effort to improve the protection of subjects enrolled into those trials. Finally, the respiratory care profession itself has benefited, owing both to its critical role in the successful implementation of complicated therapist-driven protocols and also to the ARDS Network's novel practice of utilizing respiratory therapists as clinical coordinators. This has raised the profile and enhanced the stature of the respiratory care profession. PMID- 19558742 TI - The evolution of carbon monoxide into medicine. AB - The discovery that carbon monoxide (CO)-a highly publicized toxic gas molecule can have powerful benefits and curative effects not only changed how we view CO, but has, with tremendous contradiction, resulted in clinical trials of CO for the treatment of various pathologies. There is sound preclinical evidence that, at a low concentration, CO has benefits in numerous and diverse diseases in rodents, large animals, and humans. CO especially has potential benefits in inflammatory disorders. As CO moves ahead in the clinic, we continue to advance our understanding of how it functions, especially as the number of potential clinical applications expands. CO's mechanisms of action at the cellular level depend on the disease and the experimental focus, but the one constant is that CO reestablishes homeostasis. PMID- 19558743 TI - Innovations in mechanical ventilation. AB - New features of mechanical ventilators are frequently introduced, including new modes, monitoring techniques, and triggering techniques. But new rarely translates into any measureable improvement in outcome. We describe 4 new techniques and attempt to define what is a new invention versus what is innovative-a technique that significantly improves a measurable variable. We describe and review the literature on automated weaning, automated measurement of functional residual capacity, neural triggering, and novel displays of respiratory mechanics. PMID- 19558744 TI - Extracorporeal membrane oxygenation: quo vadis? AB - Extracorporeal membrane oxygenation, a form of artificial circulatory support, continues to evolve beyond its well-established neonatal applications. It is often the most aggressive aspect of treatment algorithms in the management of severe respiratory and cardiac failure. While its use is relatively infrequent and executed in a small number of centers, it remains an important supportive measure while organ function is preserved and restored. Refinements in equipment and techniques continue to develop; patient-selection has changed, in adults and children, and cardiac applications have gained prominence. PMID- 19558745 TI - Postoperative pulmonary hypertension: etiology and treatment of a dangerous complication. AB - Postoperative pulmonary hypertension is a challenging and feared complication of many types of surgery, including lung and heart transplantation, pulmonary thromboendarterectomy, congenital-heart-disease repair, and others. The most severe manifestation is acute right heart syndrome, characterized by right heart failure and cardiovascular collapse-a daunting therapeutic challenge associated with a high mortality. Patients with postoperative pulmonary hypertension must be carefully evaluated to identify reversible contributing factors such as fluid and metabolic imbalance, hypoxemia, and right heart ischemia. A pulmonary arterial catheter and echocardiogram are recommended for evaluation, although their value has not been established in carefully designed trials. Basic principles of management include maintenance of systemic perfusion pressure, optimization of cardiac inotropy, use of lung-protective ventilator strategies, and attempting to reduce right-ventricular afterload using pulmonary vasodilators. Unfortunately, controlled trials upon which to base therapy are lacking, and most approaches are supported only by uncontrolled or anecdotal evidence. Better understanding of the pathophysiology of right heart failure and controlled trials testing therapeutic approaches are needed if we are to make progress in treating this heretofore highly mortal condition. PMID- 19558746 TI - Noninvasive ventilation in a pregnant patient with respiratory failure from all trans-retinoic-acid (ATRA) syndrome. AB - We saw a 34-year-old pregnant woman with acute promyelocytic leukemia, who developed acute respiratory failure from all-trans-retinoic acid (ATRA) syndrome. We applied noninvasive ventilation (NIV, continuous positive airway pressure plus pressure-support ventilation) to try to improve gas exchange, reduce the work of breathing, and prevent intubation. Initially we applied NIV continuously (24 hours a day), then gradually reduced the daily amount of time on NIV as her condition improved. She was discharged from the intensive care unit after 12 days. Three months after hospital discharge she gave vaginal birth to a healthy female baby. NIV was effective and safe for the mother and fetus, and NIV should be considered for respiratory failure in pregnant patients, especially if immunosuppressed. PMID- 19558747 TI - Pulmonary mass in an immunocompromised patient: think outside the box. PMID- 19558748 TI - Combining heliox and inhaled nitric oxide as rescue treatment for pulmonary interstitial emphysema. PMID- 19558750 TI - Surface-associated and secreted factors of Streptococcus suis in epidemiology, pathogenesis and vaccine development. AB - Streptococcus suis is an invasive porcine pathogen associated with meningitis, arthritis, bronchopneumonia and other diseases. The pathogen constitutes a major health problem in the swine industry worldwide. Furthermore, S. suis is an important zoonotic agent causing meningitis and other diseases in humans exposed to pigs or pork. Current knowledge on pathogenesis is limited, despite the enormous amount of data generated by 'omics' research. Accordingly, immunoprophylaxis (in pigs) is hampered by lack of a cross-protective vaccine against virulent strains of this diverse species. This review focuses on bacterial factors, both surface-associated and secreted ones, which are considered to contribute to S. suis interaction(s) with host factors and cells. Factors are presented with respect to (i) their identification and features, (ii) their distribution among S. suis and (iii) their significance for virulence, immune response and vaccination. This review also shows the enormous progress made in research on S. suis over the last few years, and it emphasizes the numerous challenging questions remaining to be answered in the future. PMID- 19558749 TI - Metabolic factors affecting the inflammatory response of periparturient dairy cows. AB - Dairy cattle are susceptible to increased incidence and severity of disease during the periparturient period. Increased health disorders have been associated with alterations in bovine immune mechanisms. Many different aspects of the bovine immune system change during the periparturient period, but uncontrolled inflammation is a dominant factor in several economically important disorders such as metritis and mastitis. In human medicine, the metabolic syndrome is known to trigger several key events that can initiate and promote uncontrolled systemic inflammation. Altered lipid metabolism, increased circulating concentrations of non-esterified fatty acids and oxidative stress are significant contributing factors to systemic inflammation and the development of inflammatory-based diseases in humans. Dairy cows undergo similar metabolic adaptations during the onset of lactation, and it was postulated that some of these physiological events may negatively impact the magnitude and duration of inflammation. This review will discuss how certain types of fatty acids may promote uncontrolled inflammation either directly or through metabolism into potent lipid mediators. The relationship of increased lipid metabolism and oxidative stress to inflammatory dysfunction will be reviewed as well. Understanding more about the underlying cause of periparturient health disorders may facilitate the design of nutritional regimens that will meet the energy requirements of cows during early lactation and reduce the susceptibility to disease as a function of compromised inflammatory responses. PMID- 19558751 TI - Bovine herpes virus infections in cattle. AB - Bovine herpes virus 1 (BHV-1) is primarily associated with clinical syndromes such as rhinotracheitis, pustular vulvovaginitis and balanoposthitis, abortion, infertility, conjunctivitis and encephalitis in bovine species. The main sources of infection are the nasal exudates and the respiratory droplets, genital secretions, semen, fetal fluids and tissues. The BHV-1 virus can become latent following a primary infection with a field isolate or vaccination with an attenuated strain. The viral genomic DNA has been demonstrated in the sensory ganglia of the trigeminal nerve in infectious bovine rhinotracheitis (IBR) and in sacral spinal ganglia in pustular vulvovaginitis and balanoposthitis cases. BHV-1 infections can be diagnosed by detection of virus or virus components and antibody by serological tests or by detection of genomic DNA by polymerase chain reaction (PCR), nucleic acid hybridization and sequencing. Inactivated vaccines and modified live virus vaccines are used for prevention of BHV-1 infections in cattle; subunit vaccines and marker vaccines are under investigation. PMID- 19558752 TI - The role of national identity representation in the relation between in-group identification and out-group derogation: ethnic versus civic representation. AB - Two studies investigated whether the content of in-group identity affects the relation between in-group identification and ethnic prejudice. The first study among university students, tested whether national identity representations (i.e., ethnic vs. civic) moderate or mediate the relation between Flemish in group identification and ethnic prejudice. A moderation hypothesis is supported when those higher in identification who subscribe to a more ethnic representation display higher ethnic prejudice levels than those higher in identification who subscribe to a more civic representation. A mediation hypothesis is supported when those higher in identification tend towards one specific representation, which in turn, should predict ethnic prejudice. Results supported a mediation hypothesis and showed that the more respondents identified with the Flemish in group, the more ethnic their identity representation, and the more they were inclined to display ethnic prejudice. The second study tested this mediation from a longitudinal perspective in a two-wave study among high school students. In group identification at Time 1 predicted over-time changes in identity representation, which in turn, predicted changes in ethnic prejudice. In addition to this, changes in identity representation were predicted by initial ethnic prejudice levels. The implications of these findings are discussed. PMID- 19558753 TI - National symbols and distinctiveness: rhetorical strategies in creating distinct national identities. AB - The purpose of this study is to examine qualitatively how respondents create national distinctiveness using rhetorical identity strategies in the context of four Finnish national symbols. The data consist of 127 essays written by Finnish secondary school students. Analysis revealed five different strategies used to distinguish between the in-group and the out-group. These strategies differ on two dimensions: the level of polarization, and the extent to which the in-group out-group relationship is depicted as being active versus passive. Furthermore, the analysis showed that the two dimensions of nationalism, particularism and universalism, have an important role in the differentiation processes and therefore highlighted the importance of taking into consideration ideological issues while studying social identities. The meaning of the contents of national identity in the differentiation processes is also discussed. PMID- 19558754 TI - Changes in student teachers' agency beliefs during a teacher education year, and relationships with observed classroom quality, and day-to-day experiences. AB - BACKGROUND: Conceptualizations of teachers' agency beliefs converge around domains of support and instruction. AIM: We investigated changes in student teachers' agency beliefs during a 1 year teacher education course, and related these to observed classroom quality and day-to-day experiences in partnership schools during the practicum. SAMPLES: Out of a sample of 66 student teachers who had responded to at least two out of four times to a questionnaire (18 men 48 women; mean age 26.4 years), 30 were observed during teaching, and 20 completed a 4-day short form diary. METHODS: Confirmatory factor analysis validated two agency belief constructs. Multi-level models for change investigated individual differences in change over time. Multi-level path models related observation and diary responses to agency beliefs. RESULTS: Supportive agency belief was high and stable across time. Instructional agency belief increased over time, suggesting a beneficial effect of teacher education. This increase was predicted by observed classroom quality (emotional support and student engagement) and daily positive affect and agency beliefs. CONCLUSIONS: Teacher education is successful in creating a context in which student teachers' supportive agency beliefs can be maintained and instructional agency beliefs can increase during the course. PMID- 19558755 TI - Effects of sad mood on autobiographical memory in older adults with and without lifetime depression. AB - OBJECTIVES: The main objective of this study was to determine whether reduced autobiographical memory specificity is also a marker for depression in older adults. To answer this question two experiments and a prospective longitudinal clinical study were executed with the autobiographical memory task (AMT) as measure for memory specificity. The objective of the 1st experimental study was to assess the influence of a negative mood induction versus the effect of multiple testing in a neutral condition in 58 never depressed (ND) elderly. In the 2nd experimental study a negative mood was induced in 63 participants remitted from depression (RD) compared to 60 matched ND controls. All participants were 55 and older. Lastly, the predictive value of the AMT for the course of depressive symptoms was assessed in the RD individuals over a 14-17 months follow-up (FU) period. RESULTS: Performance did not improve with repeated testing, but a trend was found suggesting fatigue. There was no difference between the RD and ND group on baseline AMT. The mood induction had no effect on the AMT regardless of clinical history. Changes in the level of depressive symptoms at the 14-month FU were predicted by residual symptoms at baseline and the number of previous depressive episode, but not by baseline AMT, changes in AMT or mood after mood induction. CONCLUSION: The results of this study suggest that memory specificity on the AMT may not be a useful marker for vulnerability for clinical depression in older adults. PMID- 19558756 TI - The relationship between dispositional mindfulness and conditional goal setting in depressed patients. AB - OBJECTIVES: This study explored the association between conditional goal setting (CGS; the tendency to view the achievement of high order goals for the self as conditional upon the achievement of lower order goals) and dispositional mindfulness. DESIGN/METHODS: CGS and dispositional mindfulness were assessed in 31 depressed patients in a cross-sectional design. RESULTS: A significant association was identified between increased dispositional mindfulness and reduced CGS, which remained after controlling for severity of depressed mood. CONCLUSIONS: The results suggest that treatment interventions that increase levels of mindful awareness may have the capacity to reduce CGS in depressed patients. PMID- 19558757 TI - Orthotopic neobladder versus ileal conduit urinary diversion after cystectomy--a quality-of-life based comparison. AB - INTRODUCTION: Radical cystectomy remains the gold standard in treatment of muscle invasive bladder cancer. Evolution of pathological guidelines has empowered centres to offer orthotopic substitution (OBS) to patients undergoing radical cystectomy. We compared health-related quality of life (HRQoL) between patients who underwent OBS or ileal conduit urinary diversion (ICD) following radical cystectomy. PATIENTS AND METHODS: A total of 57 patients who underwent cystectomy were assessed pre-operatively using Karnofsky performance scale (KPS). Of these, 52 patients (28 OBS and 24 ICD) who responded to a postal questionnaire consisting of SF-36 and a functional index questionnaire were included. RESULTS: Median age of patients was 70 years. Pre-operative KPS scores were similar. All eight HRQoL scales were favourable in both groups. OBS patients had significantly better physical functioning. In the cohort, 42% of men with OBS and 25% of diversions could maintain an erection to varying degrees. Of the OBS patients, 85% were continent with two patients reporting reduced QoL with pad usage. Of ICD patients, 63% felt less complete and 42% were embarrassed due to the stoma, with 58% apprehensive of stomal leakage. Of OBS patients, 96% had significant relationships and a more active life-style. CONCLUSIONS: In a similar age-group population, there was no significant difference in most QoL indices but body image issues persist in ICD patients. OBS patients had significantly better physical function, continuing to have a more active lifestyle. They attained urethral voiding with good continence. A detailed discussion of long-term functional outcome would engender a realistic expectation allowing better adaptation. PMID- 19558758 TI - How is adrenocortical cancer being managed in the UK? AB - INTRODUCTION: Adrenocortical carcinomas are rare. This case series is reported to give an overview of how adrenocortical carcinoma is currently managed in the UK. PATIENTS AND METHODS: A retrospective review was made of case notes from patients with adrenocortical carcinomas presenting to the authors (TWJL, RDB, BJH, and DS C) over the past 10 years in Newcastle, Sheffield and Cardiff. RESULTS: Newcastle treated twelve, Sheffield eleven and Cardiff seven cases. The median follow-up was 25.5 months (range, 1-102 months). All tumours were greater than 5 cm in diameter. The majority presented with symptoms of hormone excess. Adrenalectomy was performed in 83% - this was radical in 30% and followed by excision of recurrence in 13%. Adjuvant mitotane was given in 64% of patients, in combination with cytotoxic chemotherapy in 20%. One-third of patients did not receive any adjuvant therapy. There was no significant difference in survival between the three centres. The majority of patients (57%) died during the period of follow-up of this study. The median survival was 37 months (range, 2-102 months). CONCLUSIONS: The size of tumour, stage and mode of presentation, age and overall survival of patients in this study are comparable to published series of adrenocortical carcinomas from major endocrine surgical centres world-wide. Despite controversies about benefits, adjuvant mitotane was used in the majority of cases, whereas cytotoxic chemotherapy was only used in the minority. The exact role of adjuvant therapy in the management of adrenocortical carcinoma is not as well established as for other more common malignancies. Establishing a database for adrenocortical carcinomas in the UK would contribute to our understanding of the management of this disease. PMID- 19558759 TI - Audit of anticoagulation after embolectomy for acute ischaemia. AB - INTRODUCTION: Intravenous unfractionated heparin (UFH) is routinely used in patients after arterial embolectomy. Achieving and maintaining therapeutic levels requires a co-ordinated approach which may be difficult for busy junior doctors and laboratories. There is no current evidence regarding the use of subcutaneous low molecular weight heparin (LMWH) as an alternative. PATIENTS AND METHODS: The study retrospectively examined all patients who had undergone any form of embolectomy during 2006 and 2007 by review of their medical records, an electronic laboratory database, and the patients' drug charts. RESULTS: Overall, 45 patients were studied. A total of 389 activated partial thromboplastin time (APTT) tests were performed of which 146 (37.6%) were in the therapeutic range (50-90 s), 40.4% were < 50 s and 22.1% were > 90 s. Five patients (11.1%) had further surgical procedures. Significant bleeding occurred in two patients. CONCLUSIONS: The results indicate that many patients are not appropriately anticoagulated. Whilst a new UFH protocol is being developed by our hospital trust, the authors believe the use of LMWH could provide a more effective and user-friendly alternative to UFH. PMID- 19558760 TI - Clostridium difficile: how much do hospital staff know about it? AB - INTRODUCTION: The aim of this study was to determine the awareness of Clostridium difficile infection amongst healthcare professionals. SUBJECTS AND METHODS: A total of 132 healthcare professionals (18 consultants, 40 trainee doctors, and 74 nursing staff) in our hospital were randomly surveyed using a questionnaire consisting of 17 questions covering various aspects of C. difficile. RESULTS: More than 50% of healthcare professionals correctly identified C. difficile as an anaerobic bacillus. Half of the consultants and one-third of the trainees and nurses were aware that 5% of adults carry C. difficile in the gut. Overall, 80% of doctors and 40% of nursing staff (P = 0.001) were aware of the spectrum of illnesses caused by C. difficile. Seven (39%) consultants, 25 (63%) trainees, and 26 (37%) nurses correctly identified the various predisposing factors for the acquisition of C. difficile infection. Only one-third of doctors and 8% of nursing staff were aware that antibiotic restriction was the single most effective C. difficile infection control measure. In addition, 40% of doctors and 8% of nursing staff were aware that cytotoxin assay is the gold standard diagnostic test. Less than 30% of healthcare professionals were aware of the differences between the most common strain and the strain of C. difficile responsible for recent outbreaks. Only 6 (33%) consultants, 21 (53%) trainees (P = not significant), and 28 (38%) nursing staff were aware that hand washing with soap and water is the most effective way of preventing transmission of C. difficile infection. Results showed that 93% of trainees, 78% of consultants (P = 0.05) and 70% of nurses correctly answered that oral metronidazole is the drug of choice for the treatment of C. difficile infection. Compared to 73% of trainees, only two (11%) consultants (P < 0.0001) and 20 (27%) nursing staff correctly stated that oral vancomycin is the second-line treatment for persistent symptomatic C. difficile infection. CONCLUSIONS: There is a significant lack of knowledge concerning C. difficile infection amongst healthcare professions, in particular amongst consultants and nurses in our hospital. PMID- 19558761 TI - Laparoscopic appendicectomy: calculating the cost. AB - INTRODUCTION: Laparoscopic appendicectomy is a commonly performed procedure presenting a considerable cost burden. Given the additional operative costs of laparoscopic versus open appendicectomy, it is not clear whether the national tariffs are appropriate for laparoscopic appendicectomy. We conducted a study to establish the institutional costs, and to determine whether re-imbursement according to the national tariffs was sufficient. PATIENTS AND METHODS: Data were collected prospectively on patients undergoing laparoscopic appendicectomy within Leeds Teaching Hospitals Trust. Theatre and bed costs were obtained. Cost analysis was performed, and costs were compared to the re-imbursement due. RESULTS: Fifty laparoscopic appendicectomies were performed. Median operative time was 60 min. The median total operative cost of laparoscopic appendicectomy was pound906. Median equipment cost for laparoscopically completed cases was pound254. Median total in-patient cost was pound1617 (range, pound880- pound3360). This compared with a mean re-imbursement of pound1981 representing a cost benefit of pound233 per case (P = 0.0009). CONCLUSIONS: Despite a liberal use of disposable equipment, laparoscopic appendicectomy can still be performed within the confines of the national tariffs. There is a considerable variation in the cost of this procedure, and it may be possible to reduce costs by more stringent use of disposable equipment and standardising recovery protocols. PMID- 19558762 TI - A language for effective communication between surgeons and radiographers in trauma theatre. AB - INTRODUCTION: The advent of the image intensifier has revolutionised trauma surgery since its development in 1955. The manufacturers have given names to various movements of the machine in the operating manual but it has not been popular among orthopaedic surgeons or radiographers. Lack of knowledge of names of various movements and ambiguity in command often leads to confusion between the surgeon and the radiographer regarding which way to move the image intensifier. A questionnaire-based study was conducted to assess the efficacy of communication between orthopaedic surgeons and radiographers while using the image intensifier intra-operatively. SUBJECTS AND METHODS: Diagrams depicting the movements of the image intensifier were used in the questionnaire. Fifty questionnaires were given to orthopaedic surgeons and 50 to radiographers to name the various movements. RESULTS: Ninety questionnaires were returned, 45 from surgeons and 45 from radiographers. Five questionnaires from surgeons and five from radiographers were returned blank. Of those responding, 97% could name the vertical movement, 68% the horizontal movement, 12% the swivel and 29% the angulation movement. None could name the orbital movement. CONCLUSIONS: Even though orthopaedic surgeons do not operate the image intensifier themselves, knowledge of the movements of the image intensifier and their names can improve the efficacy of communication between surgeons and radiographers. A common language and precision in command can avoid confusion and has the potential to improve theatre time utilisation. The nomenclature of various movements of the image intensifier has been described and possible precise commands for various movements have been postulated. PMID- 19558763 TI - Early and long-term outcomes of patients undergoing liver resection and diaphragm excision for advanced colorectal liver metastases. AB - INTRODUCTION: At present, liver resection offers the best long-term outcome and only chance for cure in patients with colorectal liver metastases. However, there are no large series that report the early and long-term outcomes of patients who require simultaneous diaphragm excision. This study was designed to investigate these patients. PATIENTS AND METHODS: A total of 285 consecutive liver resections were performed over a 10-year period. Of these, 258 had liver resections alone and 27 underwent liver resection and simultaneous diaphragm excision. Data were collected prospectively and analysed retrospectively. Pre-operative assessment was standardised. The outcomes between the two groups were compared. RESULTS: There was no difference in age, hospital stay or intra-operative blood loss. The diaphragm was histologically involved in four out of 27 resections. As a result, the cancer involved resection margin incidence was greater in the liver resection and diaphragm excision group (14.8% versus 3.9%; P = 0.12). The median tumour size was also different between the two groups (60 mm versus 30 mm; P = 0.001). The liver and diaphragm resection group had a greater peri-operative complication rate (44.4% versus 21.3%; P = 0.02) and mortality (7.4% versus 1.6%; P = 0.25). Overall and disease-free survival was significantly worse in the group who underwent simultaneous diaphragm excision and liver resection (P = 0.04 and P = 0.005, respectively). Diaphragm invasion was found to be an independent predictor of poor overall outcome (P = 0.02). CONCLUSIONS: Liver resection and simultaneous diaphragm excision have a greater incidence of peri-operative morbidity and mortality and a significantly worse long-term outcome compared with liver resection alone. However, these data suggest that liver resection in the presence of diaphragm invasion may still offer a favourable outcome compared with chemotherapy treatment alone. Therefore, we believe that diaphragm involvement by tumour should not be a contra-indication to hepatectomy. PMID- 19558764 TI - Re-admissions following hip fracture surgery. AB - INTRODUCTION: Hip fractures in the elderly are a growing problem with a predicted incidence of 117,000 cases per year by 2016. Re-admission following a healthcare episode is an important outcome measure, which reflects non-fatal adverse events and indicates the natural history of disease. The purpose of this observational, multicentre audit was to examine rates and reasons for re-admission following hip fracture, to identify areas in the index admission and rehabilitation care that could be improved to prevent re-admission. PATIENTS AND METHODS: A total of 535 patients (> 65 years old) in two district general hospitals in the UK who underwent hip fracture surgery were recruited into the study. RESULTS: Of the study cohort, 72 patients (13.5%) died during their index admission and 88 (19.0%) of 463 patients were re-admitted once within 3 months. Causes of re admission were attributed to medical (54.8%), failure to rehabilitate (23.8%), orthopaedic (19.0%) and surgical (2.4%) reasons. Infection was the most common (31.0%) reason for re-admission and arguably the most treatable. During the 3 month postoperative period, the mortality rate was 21.3%, increasing in those re admitted to 35.1% representing the frailty of this group of patients. CONCLUSIONS: High rates of re-admission are seen following discharge in elderly patients with hip fractures. Re-admitted patients have high mortality rates. Understanding causes of re-admission may help to reduce this burden. PMID- 19558765 TI - Specialised chronic compartment measurement clinic in a district general hospital: is it worthwhile? AB - INTRODUCTION: Chronic compartment syndrome is one of the conditions that cause exertional pain in the young. In this condition, a careful history taking is usually the key for making the correct diagnosis, but intracompartmental pressure measurement that has high sensitivity and specificity rates can add invaluable confirmation, especially when surgical decompression is indicated. In a district general hospital there is usually a logistical problem in performing compartmental pressure measurement. In this study, we describe our experience in dealing with such a problem by setting up a quarterly specialised clinic for intracompartmental pressure measurement that, according to our knowledge, is unique in the UK. PATIENTS AND METHODS: We retrospectively reviewed the medical records of 42 patients referred to our specialised clinic and documented the referral patterns, clinical picture, additional investigations, pressure measurement results, final diagnosis and outcome of treatment. The intracompartmental measurement was recorded pre- and post-exercise on the treadmill for 1 min or 5 min and results were interpreted according to published criteria. RESULTS: The mean age of the referred 42 patients was 33.1 years (range, 15-78 years). There were 16 positive results (38%) confirming the diagnosis of chronic compartment syndrome and four clinically characteristic cases of this condition with negative results. There were 22 negative results (53%), one borderline, and three patients were not tested for different reasons. All of the 10 patients who were treated surgically had positive pre-operative test results. CONCLUSIONS: An inappropriate referral rate of approximately 52% was encountered that had a significant impact on the waiting time for the proposed test (5.7 months), which is far longer than the 18-week target for treatment in the NHS. We found that a positive test is a strong predictor of a successful outcome of surgical treatment. Such a specialised clinic, therefore, should only receive carefully assessed patients. PMID- 19558766 TI - Supervised exercise for intermittent claudication - an under-utilised tool. AB - INTRODUCTION: The use of supervised exercise in the management of intermittent claudication is well supported by level I evidence upon which are based grade A recommendations by the TASC II Inter-Society Consensus for the Management of Peripheral Arterial Disease and the Scottish Intercollegiate Guidelines Network (SIGN). These include that supervised exercise should be made available as part of the initial treatment for all peripheral arteriopaths. SUBJECTS AND METHODS: A questionnaire, comprising 10 questions, was drawn up to address the issues pertinent to supervised exercise in intermittent claudication. This was distributed by post, along with a pre-stamped return envelope, to all ordinary members of the Vascular Society of Great Britain and Ireland (VSGBI). All returned and received questionnaires had their responses entered onto a pre prepared spreadsheet. RESULTS: Of the 186 questionnaires posted to UK resident surgeons, 84 were returned. This equates to a response rate of 45%. Of the responders, only 24% had access to supervised exercise. There was a large spread in the proportion of eligible patients which were referred to a programme, with only 14% of VSGBI members recommending 100% of eligible patients. Rates of non compliance varied greatly. Contra-indications to supervised exercise included cardiac (27%), and vascular, musculoskeletal, geographic, and respiratory (8% each). Most supervised exercise sessions (85%) were 1 h in duration. The majority (65%) of programmes comprised one session per week. With regards the duration of programme, 55% were 3 months. Almost all classes were led by either a physiotherapist (41%) or a nurse (48%). In centres where no supervised exercise programme was available, verbal advice was given by 63%, with 34% offering leaflets. A supervised exercise set up has not been achieved due to lack of resource in 72%. CONCLUSIONS: These results are contrary to the recommendations offered by the TASC II Inter-Society Consensus and SIGN, in particular in terms of availability and referral to supervised exercise, as well as frequency of the classes where programmes were in place. The offer of information either verbally or via leaflet is commended; however, this has been shown as inferior to supervised exercise. Quoting resource as the reason for non-implementation goes against the published quality-of-life and pharmaco-economic data, which promote supervised exercise as both functionally and financially effective. This work highlights the importance of translating the results of research into evidence based clinical practice. PMID- 19558767 TI - Length of stay following primary total hip replacement. AB - INTRODUCTION: Much of the cost of primary total hip arthroplasty (THA) comprises the length of stay in hospital. Given the increasing drive for cost-effective surgery in today's National Health Service, the aim of this investigation was to determine the patient and surgical factors that most influence the length of stay following surgery. PATIENTS AND METHODS: A large, population-based study of 675 consecutive patients in a regional orthopaedic centre in the South West of Britain. RESULTS: The median length of stay was 8 days. The majority of patients (81.5%) left hospital within 2 weeks, 13.6% within 2-4 weeks and 4.9% after 4 weeks. On multivariate analysis, age above 70 years, ASA grades 3 and 4, prolonged operations and long incisions were highly significantly associated with hospital stay of over 2 weeks. CONCLUSIONS: Prolonged stay after THA is largely predetermined by case mix and this should be taken into account when units are compared for performance and in the remuneration they receive for providing this service. Slick surgery through limited incisions may reduce the length of stay. PMID- 19558769 TI - Use of the internet by patients undergoing elective hernia repair or cholecystectomy. AB - INTRODUCTION: Patient-directed information available on the internet is not always regulated; it may be confusing and sometimes just overwhelming. We aimed to establish the proportion of patients undergoing two common surgical procedures, who searched the internet for information about their operations and to assess the usefulness of the information they received. PATIENTS AND METHODS: A total of 105 consecutive patients undergoing elective abdominal wall hernia repair (n = 54) or laparoscopic cholecystectomy (n = 51) in a single surgical firm were included in the study. Patients were counselled about their operation in pre-operative assessment clinics and standard trust information leaflets were provided without any mention of this study. Patients were then asked to complete a questionnaire on the morning of their operation. RESULTS: All patients completed the questionnaire. Of the patients, 59% stated that they had access to the internet and 77% of these accessed the internet over 2 h a week. Of the patients with internet access, 31% used it to acquire additional information about their operations and 58% used internet search engines. Of the patients who searched the internet regarding their operations, 26% were confused and/or worried by the information they received. CONCLUSIONS: A significant proportion of patients undergoing common surgical procedures used the internet and about one third of them specifically sought information about their operation on the internet. Such information can cause worry and confusion in patients. Our study highlights the need for regulated, comprehensible, patient information on hospital websites to which patients should be directed. PMID- 19558768 TI - Urethral venous malformation: an unusual cause of recurrent post-coital gross haematuria in association with haematospermia. AB - Three cases of recurrent post-coital haematuria are described. Extensive protracted investigations pinpointed urethral varicosities as the likely cause. All patients were successfully treated with diathermy fulguration. PMID- 19558770 TI - Diagnosis of skin lesions by trainee surgeons: experience improves accuracy. AB - INTRODUCTION: Accurate clinical diagnosis depends on the reliable recognition of signs and symptoms. This expertise comes from experience in seeing patients which has been traditionally gained over a long training period. Shortened specialist training (Modernising Medical Careers) has led to a greater reliance on structured teaching and skills transfer programmes. The accuracy of clinical diagnosis and the rate at which diagnostic skills improve during training is important for the assessment of trainees, and the delivery of care. PATIENTS AND METHODS: This study assessed the accuracy of clinical diagnosis of skin lesions by two junior plastic surgery trainees. They were asked to diagnose 120 consecutive skin lesions seen in a pigmented skin lesion clinic in 2005, with the histological diagnosis being confirmed following subsequent excision. The process was repeated a year later in 2006 to enable the rate of correct diagnosis to be compared. RESULTS: Initially, 53.3% of diagnoses were correct. A year later, this had risen to 65.0%. Twenty-two different skin pathologies were present in excised specimens, and skin cancers comprised 30%. The trainees demonstrated 93.8% sensitivity in their initial diagnosis of malignancy (95% CI, 79.2-99.2) and 97.4% a year later (95% CI, 86.5-99.9). However, specificity was 69.3% (95% CI, 58.6-78.7) in 2005 and 71.6% (95% CI, 60.5-71.4) in 2006. CONCLUSIONS: Accuracy in the diagnosis of the wide range of skin conditions presenting to an out patient clinic was shown to increase over a 1-year period. We feel that this improvement resulted from regular clinical exposure supported by a structured learning programme. The shortening of the specialist training period may affect the acquisition of diagnostic skills by trainees and impact on the confidence with which they commence consultant practice. PMID- 19558772 TI - Cardiac stem cell therapy: advances from 2008. PMID- 19558771 TI - Improving osteoporosis assessment in the fracture clinic. AB - INTRODUCTION: The aim of this study was to compare the effectiveness of different ways of referring patients to an osteoporosis assessment service at an orthopaedic fracture clinic of a hospital in the UK. PATIENTS AND METHODS: Three methods of identifying and referring to an osteoporosis assessment service were evaluated. RESULTS: Relying on doctors for such a referral gave a catchment rate of only 1.6%. Involving patients themselves, asking them to self-refer, increased the catchment rate to 63% (P < 0.0001). Having a specialist osteoporosis and fracture liaison nurse present in clinic and reviewing the notes of patients checking in, to see if they match criteria for osteoporosis assessment, further increased catchment to 77% (P = 0.036). CONCLUSIONS: Simply having an osteoporosis assessment service and strict criteria to identify which patients should be referred to such a service will not necessarily increase catchment rate for osteoporosis patients. A nurse physically present in the clinic provided the best result, and supports the need of investing in an osteoporosis and fracture liaison nurse. PMID- 19558773 TI - Early translation of adipose-derived cell therapy for cardiovascular disease. AB - Over the past decade, cell therapy has emerged as a new approach to reversing myocardial ischemia. Several types of adult stem cells have been studied in both preclinical and clinical conditions for this purpose: bone marrow cells, circulating cells, and myoblasts. Nevertheless, the quest for the ideal "anti ischemic" cell is still ongoing. Recently, the existence of a population of stem cells located in adipose tissue (adipose-derived stem cells) has been observed. These are able to differentiate into multiple cell lineages including cardiomyocytic differentiation. In this review we discuss the basic principles of adipose-derived stem cells (types and characteristics, harvesting, and expansion), the initial experimental studies, and the currently ongoing clinical trials. PMID- 19558774 TI - Function and mode of regulation of endothelial major histocompatibility complex class II. AB - Tissue engineering is a promising approach to implement endothelial cells as a cellular delivery therapy for vascular disease. We and others previously demonstrated that endothelial cells embedded in three-dimensional collagen-based matrices retain their full biosecretory spectrum, enabling them to serve as powerful regulators of vascular diseases. Fascinatingly, matrix embedding of endothelial cells not only allows for their implantation but also seems to provide protection from allo- and xenogeneic-triggered host immune responses. This is not an effect of simple physical shielding but a more fundamental influence of cell-matrix interconnectivity on the cellular immune phenotype. Reduced cytokine-induced levels of costimulatory and adhesion molecules associated with significantly lower expression levels of major histocompatibility class II expression on matrix-embedded human aortic endothelial cells when compared to the same cells cultured on two-dimensional polystyrene coated-tissue culture plates. Strikingly, the entire interferon-gamma-dependent signaling cascade resulting in MHC class II molecule expression is markedly suppressed in endothelial cells grown to confluence within three-dimensional scaffolds. These findings might be of pivotal importance for designing endothelial cell-based therapies in general and might enhance our understanding of the underlying pathophysiology in a broad range of cardiovascular diseases (e.g., atherosclerosis, vasculitis, chronic allograft vasculopathy). PMID- 19558776 TI - Evaluation of a peritoneal-generated cardiac patch in a rat model of heterotopic heart transplantation. AB - Tissue engineering holds the promise of providing new solutions for heart transplant shortages and pediatric heart transplantation. The aim of this study was to evaluate the ability of a peritoneal-generated, tissue-engineered cardiac patch to replace damaged myocardium in a heterotopic heart transplant model. Fetal cardiac cells (1 x 10(6)/scaffold) from syngeneic Lewis rats were seeded into highly porous alginate scaffolds. The cell constructs were cultured in vitro for 4 days and then they were implanted into the rat peritoneal cavity for 1 week. During this time the peritoneal-implanted patches were vascularized and populated with myofibroblasts. They were harvested and their performance in an infrarenal heterotopic abdominal heart transplantation model was examined (n = 15). After transplantation and before reperfusion of the donor heart, a 5-mm left (n = 6) or right (n = 9) ventriculotomy was performed and the patch was sutured onto the donor heart to repair the defect. Echocardiographical studies carried out 1-2 weeks after transplantation showed normal LV function in seven of the eight hearts studied. After 1 month, visual examination of the grafted patch revealed no aneurysmal dilatation. Microscopic examination revealed, in most of the cardiac patches, a complete disappearance of the scaffold and its replacement by a consistent tissue composed of myofibroblasts embedded in collagen bundles. The cardiac patch was enriched with a relatively large number of infiltrating blood vessels. In conclusion, cardiac patches generated in the peritoneum were developed into consistent tissue patches with properties to seal and correct myocardial defects. Our study also offers a viable rat model for screening and evaluating new concepts in cardiac reconstruction and engineering. PMID- 19558775 TI - NF-kappaB activity in endothelial cells is modulated by cell substratum interactions and influences chemokine-mediated adhesion of natural killer cells. AB - Because changes in subendothelial matrix composition are associated with alterations of the endothelial immune phenotype, we sought to understand if cytokine-induced NF-kappaB activity and downstream effects depend on substrate adherence of endothelial cells (EC). We compared the upstream phosphorylation cascade, activation of NF-kappaB, and expression/secretion of downstream effects of EC grown on tissue culture polystyrene plates (TCPS) with EC embedded within collagen-based matrices (MEEC). Adhesion of natural killer (NK) cells was quantified in vitro and in vivo. NF-kappaB subunit p65 nuclear levels were significantly lower and p50 significantly higher in cytokine-stimulated MEEC than in EC-TCPS. Despite similar surface expression of TNF-alpha receptors, MEEC had significantly decreased secretion and expression of IL-6, IL-8, MCP-1, VCAM-1, and ICAM-1. Attenuated fractalkine expression and secretion in MEEC (two to threefold lower than in EC-TCPS; p < 0.0002) correlated with 3.7-fold lower NK cell adhesion to EC (6,335 +/- 420 vs. 1,735 +/- 135 cpm; p < 0.0002). Furthermore, NK cell infiltration into sites of EC implantation in vivo was significantly reduced when EC were embedded within matrix. Matrix embedding enables control of EC substratum interaction. This in turn regulates chemokine and surface molecule expression and secretion, in particular of those compounds within NF-kappaB pathways, chemoattraction of NK cells, local inflammation, and tissue repair. PMID- 19558777 TI - Implantation of a three-dimensional fibroblast matrix improves left ventricular function and blood flow after acute myocardial infarction. AB - This study was designed to determine if a viable biodegradable three-dimensional fibroblast construct (3DFC) patch implanted on the left ventricle after myocardial infarction (MI) improves left ventricular (LV) function and blood flow. We ligated the left coronary artery of adult male Sprague-Dawley rats and implanted the 3DFC at the time of the infarct. Three weeks after MI, the 3DFC improved LV systolic function by increasing (p < 0.05) ejection fraction (37 +/- 3% to 62 +/- 5%), increasing regional systolic displacement of the infarcted wall (0.04 +/- 0.02 to 0.11 +/- 0.03 cm), and shifting the passive LV diastolic pressure volume relationship toward the pressure axis. The 3FDC improved LV remodeling by decreasing (p < 0.05) LV end-systolic and end-diastolic diameters with no change in LV systolic pressure. The 3DFC did not change LV end-diastolic pressure (LV EDP; 25 +/- 2 vs. 23 +/- 2 mmHg) but the addition of captopril (2mg/L drinking water) lowered (p < 0.05) LV EDP to 12.9 +/- 2.5 mmHg and shifted the pressure-volume relationship toward the pressure axis and decreased (p < 0.05) the LV operating end-diastolic volume from 0.49 +/- 0.02 to 0.34 +/- 0.03 ml. The 3DFC increased myocardial blood flow to the infarcted anterior wall after MI over threefold (p < 0.05). This biodegradable 3DFC patch improves LV function and myocardial blood flow 3 weeks after MI. This is a potentially new approach to cell-based therapy for heart failure after MI. PMID- 19558778 TI - Percutaneous cell delivery into the heart using hydrogels polymerizing in situ. AB - Heart disease is the leading cause of death in the US. Following an acute myocardial infarction, a fibrous, noncontractile scar develops, and results in congestive heart failure in more than 500,000 patients in the US each year. Muscle regeneration and the induction of new vascular growth to treat ischemic disorders of the heart can have significant therapeutic implications. Early studies in patients with chronic ischemic systolic left ventricular dysfunction (SLVD) using skeletal myoblasts or bone marrow-derived cells report improvement in left ventricular ejection function (LVEF) and clinical status, without notable safety issues. Nonetheless, the efficacy of cell transfer for cardiovascular disease is not established, in part due to a lack of control over cell retention, survival, and function following delivery. We studied the use of biocompatible hydrogels polymerizable in situ as a cell delivery vehicle, to improve cell retention, survival, and function following delivery into the ischemic myocardium. The study was conducted using human bone marrow-derived mesenchymal stem cells and fibrin glue, but the methods are applicable to any human stem cells (adult or embryonic) and a wide range of hydrogels. We first evaluated the utility of several commercially available percutaneous catheters for delivery of viscous cell/hydrogel suspensions. Next we characterized the polymerization kinetics of fibrin glue solutions to define the ranges of concentrations compatible with catheter delivery. We then demonstrate the in vivo effectiveness of this preparation and its ability to increase cell retention and survival in a nude rat model of myocardial infarction. PMID- 19558780 TI - A transformed cell population derived from cultured mesenchymal stem cells has no functional effect after transplantation into the injured heart. AB - Bone marrow-derived mesenchymal stem cells (MSCs) are multipotent cells characterized by their self-renewal and differentiation potential. Accumulating clinical and preclinical evidence indicate MSCs are a promising cell source for regenerative medical therapies. However, undesirable immortalization, spontaneous transformation, and tumorigenic potential from long-term cultured MSCs have been reported in human and mouse. We report rat MSCs isolated from young donors could undergo transformation in early passage culture. We aimed to characterize the transformed population and determine their therapeutic effects after intracardiac transplantation in the infarcted myocardium. MSCs were isolated from bone marrow of Lewis rats according to standard protocols and cultured under standard conditions. Phenotype of growing cells was assessed by flow cytometry. Following acute myocardial infarction in rats, cells were delivered by intracardiac injection. Cardiac functions were assessed by pressure-volume loops. Infarction size and pathologic effects were evaluated after 6 weeks. The abnormal colonies were detected in culture as early at passage 3. They were noted to appear as distinctly different morphology from typical MSCs, which changed from a normal elongated spindle shape to a compact abnormal morphology. They exhibited rapid cell proliferation. Some subclones lost contact inhibition of cell division and formed multilayer aggregates. Chromosomal instability was detected. They were devoid of surface markers CD29, CD44, CD90, and CD117. Furthermore, there was no significant improvement on infarction size and cardiac function 6 weeks after cell transplantation. Our study highlights the need for establishment of biosafety criteria in regulating culture- expanded MSCs to achieve the full clinical therapeutic benefits. PMID- 19558781 TI - Myoblast transfer in ischemic heart failure: effects on rhythm stability. AB - Skeletal myoblast (SM) implantation promotes recovery of myocardial function after ischemic injury. Clinical observations suggest an association of SM implantation and ventricular arrhythmias. Support for this link has been sought in animal studies, but none employing models of congestive heart failure. In a canine model of postinfarction congestive heart failure (CHF) we compared the frequency of rhythm disturbances using ambulatory electrocardiography monitoring following skeletal myoblast or saline (SAL) implantation. In 19 mongrel dogs ischemic injury and CHF were induced by intracoronary microsphere infusions. Direct intramyocardial injection of autologous skeletal myoblasts (ASM) (2.7-8.3 x 10(8) cells) or SAL controls was administered to 11 and 8 dogs, respectively. Serial echocardiography and 24-h ambulatory electrocardiography were recorded at baseline (after CHF induction) and at 4 weeks and at 8-10 weeks after injection. Comparisons between groups of left ventricular ejection fraction (LVEF) and the frequency of ventricular arrhythmias, supraventricular arrhythmias, and measures of heart rate variability (HRV) were made at each of the three time points. LVEF increased from 41 +/- 6% to 47 +/- 2% (p < 0.03) in the ASM group, and did not change (42 +/- 6% to 40 +/- 2%, p = ns) in SAL. After injection, no differences were seen in the number of dogs demonstrating ventricular tachycardia (n = 3 vs. n = 2, p = ns) or frequent PVCs (n = 3 vs. n = 3, p = ns) in the ASM versus SAL groups, respectively. Significant changes were observed in a time-domain measure of HRV, standard deviation of normal-to-normal RR interval (in ms: 4 weeks 174 +/ 95 vs. 242 +/- 19; 8 weeks 174 +/- 78 vs. 276 +/- 78, ASM vs. SAL), but not in other time domain parameters. In this canine model of ischemic CHF, ASM implantation did not result in a significant increase in ventricular arrhythmias compared to controls animals. The potential for ASM implantation to affect time domain parameters of HRV merits further study. PMID- 19558779 TI - Ex vivo nanofiber expansion and genetic modification of human cord blood-derived progenitor/stem cells enhances vasculogenesis. AB - The stem cell therapy for treating ischemic diseases is promising; however, the limited availability and compromised quality of progenitor cells in aged and diseased patients limit its therapeutic use. Here we report a nanofiber-based ex vivo stem cell expansion technology and proangiogenic growth factors overexpression of human umbilical cord blood (UCB)-derived progenitor cells to enhance angiogenic potential of therapeutic stem cells. The progenitor cells were expanded approximately 225-fold on nanofiber-based serum-free ex vivo expansion culture technique without inducing differentiation. The expanded cells express high levels of stem cell homing receptor, CXCR4, and adhesion molecule, LFA-1. The nanofiber-expanded stem cells uptake AcLDL effectively, and migrate efficiently in an in vitro transmigration assay. These expanded cells can also differentiate into endothelial and smooth muscle cells in vitro. In a NOD/SCID mouse hind limb vascular injury model, nanofiber-expanded cells were more effective in blood flow restoration and this effect was further augmented by VEGF(164) and PDGF-BB, growth factor overexpression. The data indicate that nanofiber-based ex vivo expansion technology can provide an essential number of therapeutic stem cells. Additionally, proangiogenic growth factors overexpression in progenitor cells can potentially improve autologous or allogeneic stem cell therapy for ischemic diseases. PMID- 19558783 TI - Intramyocardial injection of autologous platelet-rich plasma combined with transmyocardial revascularization. AB - Transmyocardial revascularization (TMR) can improve refractory angina but does not consistently demonstrate an effect on myocardial function. Recent studies suggest a synergistic effect between TMR and exogenously supplied growth factors. We evaluated the clinical role of intramyocardial injection of autologous platelet-rich plasma (PRP) in conjunction with TMR. Twenty-five nonrevascularizable patients with class III/IV angina underwent minimally invasive sole therapy TMR during a 5-year period at a single institution. Group 1 (14 patients) underwent TMR alone while group 2 (11 patients) underwent TMR plus injection of PRP (Magellan plasma separator) between TMR channels. Blinded angina assessment and ejection fraction (EF) were measured preoperatively and at 6 months postoperatively. Baseline EF (57 +/- 10% vs. 50 +/- 7%), angina class (3.7 +/- 0.5 vs. 3.7 +/- 0.5), and the number of channels (48 +/- 5 vs. 48 +/- 4) were statistically similar in both groups. At 6 months, two class angina relief was similar in both groups (92% vs. 100%, p = 0.4); however, the TMR + PRP group had a lower average angina score (1.3 vs. 0.4, p = 0.07) and more were angina free (23% vs. 78%, p = 0.04) than the TMR-alone group. EF improved in the TMR + PRP group (-2.0% vs. +9.0%, p = 0.07) compared to the TMR-alone group. Two 30-day morbidities occurred in the TMR-alone group (atrial fibrillation and left pleural effusion) and one mortality occurred in the TMR + PRP group. Intramyocardial injection of autologous PRP combined with TMR may be more efficacious at relieving angina and improving myocardial function than TMR alone. PMID- 19558782 TI - Autologous bone-marrow mononuclear cell transplantation after acute myocardial infarction: comparison of two delivery techniques. AB - The objective of this study was to investigate safety and feasibility of autologous bone marrow mononuclear cells (BMMNC) transplantation in ST elevation myocardial infarction (STEMI), comparing anterograde intracoronary artery (ICA) delivery with retrograde intracoronary vein (ICV) approach. An open labeled, randomized controlled trial of 30 patients admitted with STEMI was used. Patients were enrolled if they 1) were successfully reperfused within 24 h from symptoms onset and 2) had infarct size larger than 10% of the left ventricle (LV). One hundred million BMMNC were injected in the infarct-related artery (intra-arterial group) or vein (intravenous group), 1% of which was labeled with Tc(99m) hexamethylpropylenamineoxime. Cell distribution was evaluated 4 and 24 h after injection. Baseline MRI was performed in order to evaluate microbstruction pattern. Baseline radionuclide ventriculography was performed before cell transfer and after 3 and 6 months. All the treated patients were submitted to repeat coronary angiography after 3 months. Thirty patients (57 +/- 11 years, 70% males) were randomly assigned to ICA (n = 14), ICV (n = 10), or control (n = 6) groups. No serious adverse events related to the procedure were observed. Early and late retention of radiolabeled cells was higher in the ICA than in the ICV group, independently of microcirculation obstruction. An increase of EF was observed in the ICA group (p = 0.02) compared to baseline. Injection procedures through anterograde and retrograde approaches seem to be feasible and safe. BMMNC retention by damaged heart tissue was apparently higher when the anterograde approach was used. Further studies are required to confirm these initial data. PMID- 19558784 TI - Two-dimensional speckle tracking strain analysis for efficacy assessment of myocardial cell therapy. AB - The subtle effects of transplanted bone marrow cells (BMC) on regional myocardial behavior in patients with ischemic heart disease are difficult to assess. Novel echocardiographic techniques can quantify regional myocardial deformation (strain) and distinguish between passive and active wall motion. We hypothesized that this technique may help delineate cell therapy-induced changes in regional LV contractility that escape clinical routine studies. Twelve patients with coronary artery disease and impaired LV function (LVEF &<35%) underwent CABG surgery plus intramyocardial injection of autologous bone marrow mononuclear cells. Between two and five predefined segments of ischemic myocardium per patient received BMCs, and untreated ischemic segments served as internal controls. Segmental echocardiographic analysis of peak systolic strain by speckle tracking was performed before and 1 year after surgery and compared with standard wall motion analysis. Two patients died during the follow-up period. In the remaining 10 patients, mean LVEF increased from 24.5 +/- 10% to 32.1 +/- 11% (p = 0.02). A moderate improvement of systolic function was noted in ischemic control segments by both wall motion score (WMS) and 2D strain echocardiography (2DSE). In BMC-treated segments, WMS improved slightly, but the data failed to reach statistical significance. As assessed by 2DSE, however, systolic function of BMC treated segments improved by nearly 100%. 2DSE proved to detect BMC-induced change with 30-fold higher sensitivity than WMS, and the Receiver Operating Characteristic curve (ROC) confirmed the diagnostic precision of 2DSE (area-under the-ROC = 0.87). We conclude that echocardiographic speckle tracking two dimensional strain analysis can detect cell therapy-induced changes in regional contractile function that may escape detection by standard wall motion assessment. Thus, 2DSE may be a useful tool for the further development of clinical cardiac cell therapy. PMID- 19558786 TI - Port-site pyoderma gangrenosum - a rare complication of surgical trauma. AB - We report a case of pyoderma gangrenosum occurring at the site of a laparoscopic port insertion following laparoscopic inguinal hernia repair. PMID- 19558785 TI - Non-occlusive small bowel necrosis in association with feeding jejunostomy after elective upper gastrointestinal surgery. AB - INTRODUCTION: Non-occlusive small bowel necrosis (NOSBN) has been associated with early postoperative enteral feeding. The purpose of this study was to determine the incidence of this complication in an elective upper gastrointestinal (GI) surgical patient population and the influence of both patient selection and type of feeding jejunostomy (FJ) inserted, based on the experience of two surgical units in affiliated hospitals. PATIENTS AND METHODS: The records were reviewed of 524 consecutive patients who underwent elective upper GI operations with insertion of a FJ for benign or malignant disease between 1997 and 2006. One unit routinely inserted needle catheter jejunostomies (NCJ), whilst the other selectively inserted tube jejunostomies (TJ). RESULTS: Six cases of NOSBN were identified over 120 months in 524 patients (1.15%), with no difference in incidence between routine NCJ (n = 5; 1.16%) and selective TJ (n = 1; 1.06%). Median rate of feeding at time of diagnosis was 105 ml/h (range, 75-125 ml/h), and diagnosis was made at a median of 6 days (range, 4-18 days) postoperatively. All patients developed abdominal distension, hypotension and tachycardia in the 24 h before re-exploratory laparotomy. Five patients died and one patient survived. CONCLUSIONS: The understanding of the pathophysiology of NOSBN is still rudimentary; nevertheless, its 1% incidence in the present study does call into question its routine postoperative use especially in those at high risk with an open abdomen, planned repeat laparotomies or marked bowel oedema. Patients should be fully resuscitated before initiating any enteral feeding, and feeding should be interrupted if there is any evidence of feed intolerance. PMID- 19558787 TI - Introduction of a day-case laparoscopic cholecystectomy service in the UK: a critical analysis of factors influencing same-day discharge and contact with primary care providers. AB - INTRODUCTION: The objective of this study was to determine the safety and acceptability of the implementation of a day-case laparoscopic cholecystectomy (LC) service in a large UK teaching hospital, and analyse factors influencing contact with primary care providers. Wide-spread introduction of day-case LC in the UK is a major target of healthcare providers. However, few centres have reported their experience. In the US, out-patient surgery for LC has been reported, though many groups have utilised 24-h observation units to facilitate discharge. Concerns remain amongst surgeons regarding the feasibility and acceptability of the introduction of day-case LC in the UK. PATIENTS AND METHODS: Comprehensive care and operative data were prospectively collected on the first 106 consecutive day-case procedures in our hospital. Postoperative recovery was monitored by telephone questionnaire on days 2, 5 and 14, including complications, satisfaction and general practitioner consultation. RESULTS: A total of 106 patients were admitted for day-case LC, of whom 84% were discharged on the day of surgery. Patient satisfaction rate was 94% in both the successful day-case and the admitted patients. Mean operation time was 62 min, with an average total stay on the day-care unit of 426 min. Training-grade surgeons performed 31% of operations. Both the readmission rate after surgery and rate of conversion to open surgery were 2%. Advice from primary healthcare providers was sought by 33% of patients within the first 14 postoperative days. CONCLUSIONS: Introduction of day-case LC in the UK is feasible and acceptable to patients. The potential burden to primary care providers needs further study. PMID- 19558788 TI - Next-generation gene targeting in the mouse for functional genomics. AB - In order to elucidate ultimate biological function of the genome, the model animal system carrying mutations is indispensable. Recently, large-scale mutagenesis projects have been launched in various species. Especially, the mouse is considered to be an ideal model to human because it is a mammalian species accompanied with well-established genetic as well as embryonic technologies. In 1990's, large-scale mouse mutagenesis projects firstly initiated with a potent chemical mutagen, N-ethyl-N-nitrosourea (ENU) by the phenotype-driven approach or forward genetics. The knockout mouse mutagenesis projects with trapping/conditional mutagenesis have then followed as Phase II since 2006 by the gene-driven approach or reverse genetics. Recently, the next-generation gene targeting system has also become available to the research community, which allows us to establish and analyze mutant mice carrying an allelic series of base substitutions in target genes as another reverse genetics. Overall trends in the large-scale mouse mutagenesis will be reviewed in this article particularly focusing on the new advancement of the next-generation gene targeting system. The drastic expansion of the mutant mouse resources altogether will enhance the systematic understanding of the life. The construction of the mutant mouse resources developed by the forward and reverse genetic mutagenesis is just the beginning of the annotation of mammalian genome. They provide basic infrastructure to understand the molecular mechanism of the gene and genome and will contribute to not only basic researches but also applied sciences such as human disease modelling, genomic medicine and personalized medicine. PMID- 19558789 TI - Autophagy in neurodegeneration: two sides of the same coin. AB - Autophagy is a bulk lysosomal degradation process important in development, differentiation and cellular homeostasis in multiple organs. Interestingly, neuronal survival is highly dependent on autophagy due to its post-mitotic nature, polarized morphology and active protein trafficking. A growing body of evidence now suggests that alteration or dysfunction of autophagy causes accumulation of abnormal proteins and/or damaged organelles, thereby leading to neurodegenerative disease. Although autophagy generally prevents neuronal cell death, it plays a protective or detrimental role in neurodegenerative disease depending on the environment. In this review, the two sides of autophagy will be discussed in the context of several neurodegenerative diseases. PMID- 19558790 TI - Glycosylation modification of human prion protein provokes apoptosis in HeLa cells in vitro. AB - We investigate the correlation between the glycosylation modified prion proteins and apoptosis. The wild-type PRNP gene and four PRNP gene glycosylated mutants were transiently expressed in HeLa cells. The effect of apoptosis induced by PrP mutants was confirmed by MTT assay, Hochest staining, Annexin-V staining and PI staining. ROS test detected ROS generation within the cells. The mitochondrial membrane potential was analyzed by the flow cytometry. The expression levels of Bcl-xL, Bax, cleaved Caspase-9 proteins were analyzed by Western Blot. The results indicated that the expressed non-glycosylated PrP in HeLa cells obviously induced apoptosis, inhibited the growth of cells and reduced the mitochondrial membrane potential, and more ROS generation and low levels of the apoptosis related proteins Bcl-xL, the activated the cleaved Caspase-9 proteins were found. The apoptosis induced by non-glycosylated PrP demonstrates that its underlying mechanism correlates with the mitochondria-mediated signal transduction pathway. PMID- 19558791 TI - Beta-catenin protein utilized by Tumour necrosis factor-alpha in porcine preadipocytes to suppress differentiation. AB - The Wnt/beta-catenin signaling pathway alters adipocyte differentiation by inhibiting adipogenic gene expression. beta-catenin plays a central role in the Wnt/beta-catenin signaling pathway. In this study, we revealed that tumour necrosis factor-alpha (TNF-alpha), a potential negative regulator of adipocyte differentiation, inhibits porcine adipogenesis through activation of the Wnt/beta catenin signaling pathway. Under the optimal concentration of TNF-alpha, the intracellular beta-catenin protein was stabilized. Thus, the intracellular lipid accumulation of porcine preadipocyte was suppressed and the expression of important adipocyte marker genes, including peroxisome proliferator-activated receptor-gamma (PPARgamma) and CCAAT/enhancer binding protein-alpha (C/EBPalpha), were inhibited. However, a loss of beta-catenin in porcine preadipocytes enhanced the adipogenic differentiation and attenuated TNF-alpha induced anti adipogenesis. Taken together, this study indicated that TNF-alpha inhibits adipogenesis through stabilization of beta-catenin protein in porcine preadipocytes. PMID- 19558792 TI - Keratin 17 identified by proteomic analysis may be involved in tumor angiogenesis. AB - Angiogenesis is crucial for solid tumor growth. By secreting angiogenic factors, tumor cells induce angiogenesis. However, targeting these angiogenic factors for cancer therapy is not always successful, suggesting that other factors may be involved in tumor angiogenesis. This work shows that 25 protein spots were differentially expressed by two-dimensional gel electrophoretic analysis when HepG2 cells induced endothelial cell differentiation to tube in vitro, and most of them were upregulated. Twenty-one proteins were identified with MALDI-TOF-MS, and the other four were identified by LTQ-MS/MS. Keratins were identified as one class of these upregulated proteins. Further study indicated that the expression of keratin 17 in cultured endothelial cells is likely microenvironment regulated, because its expression can be induced by HepG2 cells and bFGF as well as serum in culture media. Increased expression of keratins in endothelial cells, such as keratin 17, may contribute to the angiogenesis induced by HepG2 cells. PMID- 19558793 TI - Enzymatic properties of the N- and C-terminal halves of human hexokinase II. AB - Although previous studies on hexokinase (HK) II indicate both the N- and C terminal halves are catalytically active, we show in this study the N-terminal half is significantly more catalytic than the C-terminal half in addition to having a significantly higher Km for ATP and Glu. Furthermore, truncated forms of intact HK II lacking its first N-terminal 18 amino acids (delta18) and a truncated N-terminal half lacking its first 18 amino acids (delta18N) have higher catalytic activity than other mutants tested. Similar results were obtained by PET-scan analysis using (18)FFDG. Our results collectively suggest that each domain of HK II possesses enzyme activity, unlike HK I, with the N-terminal half showing higher enzyme activity than the C-terminal half. PMID- 19558794 TI - Human intronless disease associated genes are slowly evolving. AB - In the present study we have examined human-mouse homologous intronless disease and non-disease genes alongside their extent of sequence conservation, tissue expression, domain and gene ontology composition to get an idea regarding evolutionary and functional attributes. We show that selection has significantly discriminated between the two groups and the disease associated genes in particular exhibit lower K(a) and K(a)/K(s) while K(s) although smaller is not significantly different. Our analyses suggest that majority of disease related intronless human genes have homology limited to eukaryotic genomes and their expression is localized. Also we observed that different classes of intronless disease related genes have experienced diverse selective pressures and are enriched for higher level functionality that is essentially needed for developmental processes in complex organisms. It is expected that these insights will enhance our understanding of the nature of these genes and also improve our ability to identify disease related intronless genes. PMID- 19558795 TI - The subcellular distribution of MnSOD alters during sodium selenite-induced apoptosis. AB - It was reported that high doses of sodium selenite can induce apoptosis of cancer cells, but the molecular mechanisms are poorly understood. Manganese superoxide dismutase (MnSOD) converts superoxide radical to hydrogen peroxide within the mitochondrial matrix and is one of the most important antioxidant enzymes. In this study, we showed that 20 microM sodium selenite could alter subcellular distribution of MnSOD, namely a decrease in mitochondria and an increase in cytosol. The alteration of subcellular distribution of MnSOD is dependent on the production of superoxide induced by sodium selenite. PMID- 19558796 TI - Biochemical characterization of the lipid-binding properties of a broccoli cuticular wax-associated protein, WAX9D, and its application. AB - In this study, we showed that WAX9D, a nonspecific lipid-transfer protein found in broccoli, binds palmitate (C16) and stearate (C18) with dissociation constants of 0.56 muM and 0.52 muM, respectively. WAX9D was fused to thioredoxin protein by genetic manipulation to enhance its solubility. The data revealed strong interaction of Trx-WAX9D with palmitate and stearate. The dissociation constants of Trx-WAX9D for palmitate and stearate were 1.1 muM and 6.4 muM, respectively. The calculated number of binding sites for palmitate and stearate was 2.5 to 2.7, indicating that Trx-WAX9D can bind three molecules of fatty acids. Additionally, Trx-WAX9D was shown to inhibit the apoptotic effect of palmitate in endothelial cells. Our data using Trx-WAX9D provide insight into the broad spectrum of its biological applications with specific palmitate binding. PMID- 19558797 TI - Synthesis and base pairing properties of DNA-RNA heteroduplex containing 5 hydroxyuridine. AB - 5-Hydroxyuridine (5-OHU) is a major lesion of uridine and cytosine produced in RNA by various chemical oxidants. To elucidate its biochemical and biophysical effects on RNA replication, the site-specifically modified oligoribonucleotides containing 5-OHU were synthesized with C5-hydroxy-5'-O-DMTr-2'-TBDMS-uridine phosphoramidite using automated solid phase synthesis. The base-pairing properties of nucleotides opposite 5-OHU in 24 mer oligoribonulcleotides with dNTP were studied using three reverse transcriptases (Super-Script(TM)II-, AMV-, MMLV-RT) in cDNA synthesis. Adenine as well as guanine was incorporated preferentially by all reverse transcriptases. In the UV-melting temperature experiment, the results from the relative stabilities of the base pairs were A : 5-OHU > G : 5-OHU > T : 5-OHU approximately C : 5-OHU. Circular Dichroism (CD) studies showed that DNA-RNA containing 5-OHU heteroduplexes exhibit a similar conformation between the A-type RNA and B-type DNA. These results suggest that 5 OHU from oxidative damage was mainly influenced by adenine mismatch. PMID- 19558798 TI - NSA9, a human prothrombin kringle-2-derived peptide, acts as an inhibitor of kringle-2-induced activation in EOC2 microglia. AB - In neurodegenerative diseases, such as Alzheimer's and Parkinson's, microglial cell activation is thought to contribute to CNS injury by producing neurotoxic compounds. Prothrombin and kringle-2 increase levels of NO and the mRNA expression of iNOS, IL-1beta, and TNF-alpha in microglial cells. In contrast, the human prothrombin kringle-2 derived peptide NSA9 inhibits NO release and the production of pro-inflammatory cytokines such as IL-1beta, TNF-alpha, and IL-6 in LPS-activated EOC2 microglia. In this study, we investigated the anti inflammatory effects of NSA9 in human prothrombin- and kringle-2-stimulated EOC2 microglia. Treatment with 20-100 muM of NSA9 attenuated both prothrombin- and kringle-2-induced microglial activation. NO production induced by MAPKs and NF kappaB was similarly reduced by inhibitors of ERK (PD98059), p38 (SB203580), NF kappaB (N-acetylcysteine), and NSA9. These results suggest that NSA9 acts independently as an inhibitor of microglial activation and that its effects in EOC2 microglia are not influenced by the presence of kringle-2. PMID- 19558799 TI - HP0902 from Helicobacter pylori is a thermostable, dimeric protein belonging to an all-beta topology of the cupin superfamily. AB - Here, we report the first biochemical and structural characterization of the hypothetical protein HP0902 from Helicobacter pylori, in terms of structural genomics. Gel-permeation chromatography and dynamic light scattering indicated that the protein behaves as a dimer in solution. Circular dichroism spectroscopy showed that HP0902 primarily adopts a beta-structure and the protein was highly thermostable with a denaturing temperature higher than 70 degrees C. Finally, the backbone NMR assignments were obtained on the [(13)C,(15)N]HP0902 and the secondary structure was determined using the chemical shift data. Additionally, the local flexibility was assessed via a heteronuclear (1)H-(15)N steady state NOE experiment. The results revealed that HP0902 would adopt a compactly folded, all-beta topology with 11 beta-strands. All of the results clearly support the notion that HP0902 belongs to the cupin superfamily of proteins. PMID- 19558800 TI - Gene polymorphism of vascular endothelial growth factor in children with Henoch Schonlein purpura nephritis. AB - OBJECTIVE: To study the relationship of -634G/C gene polymorphism of vascular endothelial growth factor (VEGF) with Henoch-Schonlein purpura nephritis (HSPN) in children. METHODS: One hundred ethnic Han children with HSP, including 50 children with concurrent nephritis (HSPN group) and 50 children without nephritis (HSP without nephritis group), were enrolled. Fifty age-, sex-and ethnics-matched healthy children were used as the control group. VEGF-634G/C genotypes were determined by PCR-RFLP. Plasma VEGF levels were measured using ELISA. RESULTS: CC genotype distribution (32%) and C allele frequency (56%) in the HSPN group were significantly higher than those in the control group (10% and 35% respectively) and the HSP without nephritis group (10% and 33% respectively) (P<0.01). The incidence of nephritis in HSP patients with CC genotype increased significantly when compared with those with GG genotype (76% vs 31%; P<0.01). Plasma VEGF levels in patients with CC genotype (180.5+/- 40.7 pg/mL) were significantly higher than those in patients with CG (145.2+/- 48.3 pg/mL) and GG (101.5+/- 26.5 pg/mL) genotypes (P<0.05). CONCLUSIONS: VEGF-634G/C gene polymorphism may be associated with the development of HSPN. C allele may a susceptible gene of HSPN. PMID- 19558801 TI - [Investigation of campylobacter jejuni infection in children with diarrhea in Guangzhou]. AB - OBJECTIVE: To investigate the incidence of campylobacter jejuni (CJ) infection and the drug resistance of CJ in children with diarrhea in Guangzhou. METHODS: The fecal samples of 3,351 children with diarrhea between July 2005 and June 2008 were collected for CJ culture. The species of CJ strains were identified by Lior methods. The drug susceptibility tests were performed by the Kirby-Bauer method. RESULTS: Two hundred and sixty-seven CJ strains (8.0%) were isolated from 3,351 samples. The children at age of 1 month to 1 year were susceptible to CJ, accounting for 91.0%. A higher incidence of CJ infection (76.8%) was found in summer and autumn. The CJ strains were susceptible to imipenem, amikacin, cefoperazone/sulbactam, chloramphenicol, macrolides and lincomycins. Parts of CJ strains (20%-40%) were resistant to ampicillin, quinolones and ambramycin. All CJ strains were resistant to sulfamethoxazole/trimethoprim and cefditoren. Two hundred and one strains (75.3%) were CJ biotype I. CONCLUSIONS: CJ is an important pathogen of diarrhea in children from Guangzhou. CJ is resistant to some antibiotics used often in clinical practice, and so it is thus important to use antibiotics based on the results of drug susceptibility tests in children with CJ infection. PMID- 19558802 TI - [Role of oxidative stress in the pathogenesis of esophageal mucosal injury in children with reflux esophagitis]. AB - OBJECTIVE: To investigate the role of oxidative stress in the pathogenesis of esophageal mucosa injury in children with reflux esophagitis (RE). METHODS: Esophageal mucosal samples from 36 children with RE (7 months to 16 years of age) were obtained by gastroscopy. The parameters of oxidative stress, including the contents of malondialdehyde (MDA), glutathione (GSH) and nitric oxide (NO) and total superoxide dismutase (T-SOD) activity in the esophageal mucosa as well as the protein content of the esophageal mucosa, were measured. Twenty children (3 to 16 years of age) without esophageal mucosal injury by gastroscopy served as controls. RESULTS: There was no significant difference in the protein content of the esophageal mucosa between the RE and the control groups. The content of MDA in the RE group (15.36+/- 16.67 nmol/mg) was significantly higher than that in the control group (7.51+/- 6.17 nmol/mg) (P<0.01). The activity of T-SOD in the RE group (30.43+/- 35.09 U/mg) was statistically lower than that in the control group (56.34+/- 51.73 U/mg) (P<0.05). No significant differences were observed in GSH and NO contents between the two groups. CONCLUSIONS: The MDA content increases and the SOD content decreases in the esophageal mucosa in children with RE. This suggests that oxidative stress seems to be an important mediator in generation of esophageal mucosal injury. PMID- 19558803 TI - [Values of brain natriuretic peptide and N-terminal pro-brain natriuretic peptide in evaluation of cardiac function in children with congenital heart disease]. AB - OBJECTIVE: To study the values of brain natriuretic peptide (BNP) and N-terminal pro-brain natriuretic peptide (NT-proBNP) in the evaluation of cardiac function in children with congenital heart disease (CHD). METHODS: Seventy-one children with CHD were classified to two groups: congestive heart failure (CHF) (n=23 ) and non-CHF (n=48). Thirty-five age-matched normal children were used as the control group. Plasma BNP content was measured using a microparticle enzyme immunoassay (MEIA) on the AxSYM. Plasma NT-proBNP content was measured using an automated electrochemiluminescence immunoassay on a Roche Modular Analytics E170 analyzer. Echocardiographic parameters, including left ventricular end diastolic dimension index (LVEDDI) and left ventricular ejection fraction (LVEF), were measured. RESULTS: Plasma BNP and NT-proBNP contents in the CHF group were significantly higher than those in the non-CHF group (P<0.01). The non-CHF group had higher plasma BNP and NT-proBNP contents than the control group (P<0.01). LogBNP and LogNT-proBNP values were negatively correlated with the LVEF in the CHF group (r=-0.64, r=-0.67 respectively, P<0.01), and they were positively correlated with the LVEDDI (r=0.58, r=0.76 respectively, P<0.01). In the non-CHF group, LogBNP and LogNT-proBNP values were not correlated with the LVEF, but a positive correlation was found between the LogNT-proBNP value and the LVEDDI (r=0.35, P<0.05). Using plasma BNP content > or =149.8 pg/mL and NT-proBNP content > or =820.1 pg/mL as cut-off values for diagnosing CHF respectively, the sensitivities were 87.0 % and 91.3% respectively, the specificities were 91.7% and 97.9% respectively, and the areas under the ROC curves were 0.935 and 0.987 respectively. CONCLUSIONS: Both BNP and NT-proBNP can be useful in assessment of cardiac function and diagnosis of CHF in children with CHD. NT-proBNP appears to be more sensitive and specific in the diagnosis of CHF than BNP. PMID- 19558805 TI - [Qualitative research on the family management and family needs of children with asthma]. AB - OBJECTIVE: A qualitative research was conducted to investigate the problems on the family management of asthmatic children and the needs for family health services in order to provide basis for family-centered care. METHODS: Fifteen caregivers of children with asthma were interviewed with open-ended questions. The collected data were studied using Colaizzi's seven-step method of phenomenological analysis. RESULTS: The problems of family management and the needs for family health services were shown as follows: insufficient knowledge to prevention and treatment of asthma, poor compliance, ignoring psychological effects of asthma on children, a family's failure to cope with the distress and financial burden. CONCLUSIONS: It is important to provide asthma education and prevention program for caregivers and encourage them to participate in the design of medical program for asthmatic children. Individual asthma education and guides are also necessary for caregivers. PMID- 19558804 TI - [Application of neurally adjusted ventilatory assist in infants who underwent cardiac surgery for congenital heart disease]. AB - OBJECTIVE: Neurally adjusted ventilatory assist (NAVA) is a new mode of mechanical ventilation that delivers ventilatory assist in proportion to neural effort. This study aimed to compare the hemodynamic safety, oxygenation and gas exchange effects ventilated with NAVA and with pressure support ventilation (PSV) in infants who underwent open-heart surgery. METHODS: Twenty-one infants who underwent open-heart surgery for congenital heart disease (mean age 2.9+/- 2.1 months and mean weight 4.2+/- 1.4 kg) were enrolled. They were ventilated with PSV and NAVA for 60 minutes respectively in a randomized order. The hemodynamic, oxygenation and gas exchange effects produced by the two ventilation modes were compared. RESULTS: Three cases failed to shift to NAVA because of the bilateral diaphragmatic paralysis after operation. In the other 18 cases, there were no significant differences in the heart rate (HR), systolic blood pressure (BPs) and central venous pressure (CVP) in the two ventilation modes. The PaO2/FiO2 (P/F) ratio in NAVA was slightly higher than in PSV, but there was no statistical difference. PaCO2 did not show significant differences in the two modes. The peak inspiratory pressure (PIP) and electrical activity of the diaphragm (EAdi) in NAVA were significantly lower than in PSV. The EAdi signal after extubation was higher in infants who needed reintubation or intervention of noninvasive mechanical ventilation than in those who were extubated successfully (30.0+/- 8.4 microV vs 11.1+/- 3.6 microV; P<0.01). CONCLUSIONS: As the first study of application of NAVA in infants in China, this study shows that NAVA has the same homodynamic effects as PSV. However the PIP for maintaining the same level of PaCO2 in NAVA is significantly lower than that in the traditional PSV. Monitoring the EAdi signal after extubation may show the risks of reintubation or intervention of noninvasive mechanical ventilation. PMID- 19558806 TI - [Relationship between the efficacy of inhaled corticosteroids and the leukotriene expression in children with asthma]. AB - OBJECTIVE: To evaluate the relationship between leukotriene expression in blood polymorphonuclear leukocytes (PMNL) and the efficacy of inhaled corticosteroids (ICS) in children with asthma. METHODS: Thirty-two children with asthma (5-12 years) and ten healthy children (control group) were enrolled. The asthmatic children were subdivided into ICS well-controlled and ICS poorly-controlled groups based on their clinical symptoms and lung function. The level of leukotriene C4 synthase (LTC4S) mRNA in PMNL was detected by fluorescence quantitative polymerase chain reaction. The level of LTC4S mRNA was expressed by the value of qCt, and the value of qCt was diversely correlated with the level of LTC4S mRNA expression. The concentration of urinary leukotriene E4 (LTE4) was measured using ELISA. RESULTS: The expression of LTC4S mRNA in PMNL was significantly higher in children with asthma (qCt: 1.12+/-0.27) than that in the control group (qCt: 1.42+/-0.12; P< 0.05). The expression of LTC4S mRNA in PMNL in the ICS poorly-controlled group (qCt: 1.03+/-0.17) was significantly higher than that in the ICS well-controlled group (qCt: 1.24+/-0.33; P< 0.05) and the control group(1.42+/-0.12; P< 0.01). There was no significant difference in the level of urinary LTE4 among the the ICS poorly-controlled, the ICS well controlled and the control groups. CONCLUSIONS: LTC4S mRNA expression in PMNL in asthmatic children increases, and the LTC4S mRNA expression in the ICS poorly controlled group is higher than that in the ICS well-controlled group. This suggests that an increased leukotriene expression might be associated with poorly controlled asthma. PMID- 19558807 TI - [A comparative study of conventional dose and low dose adrenocorticotrophic hormone therapy for West syndrome]. AB - OBJECTIVE: The efficacy and adverse effects of conventional dose and low dose adrenocorticotrophic hormone (ACTH) therapy for West syndrome (WS) were compared in order to identify a low effective dose with few adverse effects. METHODS: A prospective randomized controlled study was conducted. Thirty children with cryptogenic (n=8) or symptomatic (n=22) WS were enrolled. They were randomly assigned to receive either conventional dose or low dose ACTH therapy. For the conventional dose group, ACTH 50 IU per day was administered for 2 weeks and tapered to zero over the subsequent 2 weeks. For the low dose group, 0.4 IU/kg per day was injected for 2 weeks. After seizures were fully controlled, ACTH was tapered to zero over the subsequent 2 weeks. If there was an absence of an effective response in the low dose group, the dosage was increased to 1 IU/kg per day for the next 2 weeks and then tapered to zero over 2 weeks. Both effectiveness and adverse effects were compared between the two groups. RESULTS: There were no significant differences in the good initial responses between the conventional and the low dose groups, which were 53% and 60%, respectively (P> 0.05). EEG findings after ACTH therapy, the rate of relapse of spasms, and the interval to relapse were not different between the two groups (P> 0.05). The long term outcomes were assessed in the initial 8 responders, and there were no significant differences between the two groups (follow-up duration>12 months). The rates of good efficacy and disappearance of the hypsarrhythmia were significantly higher in the cryptogenic WS group than in the symptomatic WS group (P<0.05). The incidence of ACTH therapy related-adverse effects in the conventional dose group (93%) was significantly higher than in the low dose group (20%) (P<0.01). The mild brain shrinkage was observed in one patient from the conventional dose group. CONCLUSIONS: The short-term and long-term therapeutic effects of ACTH between 50 IU/d and 0.4 IU/kg/d doses are similar. ACTH therapy is more effective for cryptogenic WS than symptomatic WS. To reduce adverse effects, ACTH therapy should start with a low dose (0.4 IU/ kg each day). PMID- 19558808 TI - [hTERT gene expression in children with beta-thalassemia major]. AB - OBJECTIVE: Human telomerase reverse transcriptase (hTERT) is a rate-limiting enzyme which dictates the activity of human telomerase and thus decides the life span of cells. The aim of this study was to explore the expression of hTERT in bone marrow from children with beta-thalassemia major and the relationship between the expression of hTERT and hemoglobin levels. METHODS: Multiple allele specific polymerase chain reaction (MASPCR) was used for targeted DNA amplification and gene mutation analysis of beta-thalassemia. hTERT mRNA expression in bone marrow was examined using real-time reverse transcription polymerase chain reaction (RT-PCR) analysis in 29 children with beta-thalassemia major, in 10 children with agranulocytosis and in K562 cell line. The hemoglobin levels in peripheral blood were measured. The relationship between hTERT expression and hemoglobin levels was evaluated by the Spearman test in the beta thalassemia major group. RESULTS: hTERT mRNA expression significantly increased in bone marrow from children with beta-thalassemia major compared with that from children with agranulocytosis (0.2928+/- 0.0838 vs 0.0993+/- 0.0336; P<0.01), but was significantly lower than that in K562 cell line (0.8291+/- 0.0908) (P<0.01). A significantly inverse correlation was found between hTERT mRNA expression and hemoglobin levels (r=-0.841, P<0.01). CONCLUSIONS: A low hemoglobin concentration might contribute to the up-regulation of marrow hTERT expression in children with beta-thalassemia major. PMID- 19558809 TI - [Expression of heat shock protein 90 in peripheral blood mononuclear cells of children with myasthenia gravis]. AB - OBJECTIVE: Myasthenia gravis is an autoimmunity disease and its pathogenesis has not been fully identified. Heat shock protein 90 (HSP90) shows an abnormal expression in other autoimmunity diseases. This study examined the mRNA expression of two isoforms of HSP90 in peripheral blood mononuclear cells (PBMC) and serum cortisol content in children with myasthenia gravis. METHODS: Thirty six children with myasthenia gravis and 19 healthy children were enrolled. Serum cortisol content was measured by the chemiluminescence assay. The expression of HSP90alpha and HSP90beta mRNA in PBMC was detected by the RT-PCR technique. RESULTS: The mRNA expression of HSP90alpha (0.7329+/- 0.2120) and HSP90beta (0.7193+/- 0.2869) in children with myasthenia gravis was significantly higher than that in healthy controls (0.5574+/- 0.2084 and 0.4892+/- 0.2104 respectively) (P<0.01). Serum cortisol content (285.04+/- 146.39 nmol/L) in children with myasthenia gravis was also higher than that in the healthy controls (196.25+/- 64.52 nmol/L) (P<0.01). CONCLUSIONS: The high mRNA expression of HSP90alpha and HSP90beta in PBMC might be associated with the development of myasthenia gravis. The high serum cortisol level indicates a high stress state or might be correlated to the glucocorticoid receptor abnormality in children with myasthenia gravis. PMID- 19558810 TI - [Treatment and outcome of 14 children with hepatoblastoma]. AB - OBJECTIVE: To assess the outcome of childhood hepatoblastoma after a combination therapy of resection and chemotherapy. METHODS: The clinical data of 14 children with hepatoblastoma was retrospectively reviewed. Their long-term survival was followed-up. RESULTS: Twelve cases received surgery and planned chemotherapy. The follow-up duration averaged 18 months (range 1.5-74 months). Nine survived free of events, 1 died, 1 survived with multiple lung metastases, and 1 with increased alpha-fetoprotein (AFP) content but without residual tumor. CONCLUSIONS: Surgery assisted with chemotherapy can improve the outcome of hepatoblastoma. PMID- 19558811 TI - [Survey of feeding patterns in 437 infants from Huining County of Gansu Province]. AB - OBJECTIVE: To investigate the feeding patterns of infants in Huining County, Gansu Province, an economically undeveloped county in China, so as to provide scientific evidence for nutritional intervention. METHODS: By cluster sampling based on the area distribution and the economic state, 437 children from 7 different areas in Huining County, Gansu Province were enrolled. A questionnaire on the feeding patterns within 24 months of age was completed by the caregivers. The addition of nutrient supplements (calcium, zinc, ferri, vitamin AD, etc.) during the last year was investigated using the food frequency questionnaires. RESULTS: The average percentages of breast feeding, mixed feeding and artificial feeding were 45.3%, 40.5% and 14.2% respectively. Delayed introduction of complementary food was the most common. Formula powder was added earliest, at age of 6.4 months. The meats were added latest, at age of 17.2 months. Only 66.8% of infants received an addition of complementary food at age of 4-6 months. Grains, vegetables, fruits and eggs were common complementary additional diets, but fishes and shrimps were seldom provided. The proportion of nutrient supplement use was low. The choice of nutrient supplements in urban districts mainly depended on the physicians' suggestions, but in rural area it depended on the caregivers' experience and media advertisements. CONCLUSIONS: The feeding patterns for the infants are not reasonable in Huining County, Gansu Province. It is important to provide nutritional education and reasonable feeding program for caregivers from economically undeveloped areas. PMID- 19558812 TI - [Protective effects of delayed multiple course hyperbaric oxygen treatment against hypoxic-ischemic brain damage in neonatal rats]. AB - OBJECTIVE: To study the protective effects of multiple course hyperbaric oxygen (HBO) treatment against hypoxic-ischemic brain damage (HIBD) in neonatal rats when HBO treatment is delayed (96 hrs after the HIBD event). METHODS: Eighty eight 7-day-old Sprague-Dawley rat pups were randomly assigned to control, HIBD and HBO groups. The HBO group was subdivided into cohorts receiving treatment 2 h, 48 h and 96 h, respectively, after HIBD was induced. The three subgroups comprising different therapeutic windows were further randomly assigned to receive 1, 2 or 3 courses of HBO treatment ("HBO-1, -2 and -3 sub-groups"). HBO was administered once daily (2 ATA), a course lasting for seven days. There was an interval of three days between the courses. All pups were sacrificed at the end of HBO treatment (31 days after HIBD). TUNEL staining was used for testing neuronal apoptosis in the cortex and the CA1 of the hippocampus, and NSE staining was used to ascertain cortical neuronal population. RESULTS: 1.There were significantly more TUNEL positive cells in the HIBD group than in the control group; NSE positive cells were significantly lower than in controls (P<0.01). 2. With the more delayed therapeutic window, the effects of apoptosis inhibition and neuronal protection of a single course of HBO were gradually reduced. 3. With increasing courses of HBO treatment, the effects of apoptosis inhibition and neuronal protection of HBO increased gradually in rats receiving treatment 48 and 96 hrs after HIBD. In the HBO group receiving treatment 2 hrs after HIBD, the number of apoptotic cells and NSE positive cells were close to that of the control group after one course of HBO treatment. CONCLUSIONS: One course of HBO administered within 2 hrs after HIBD can effectively inhibit neuron apoptosis and protect neurons. The effects of apoptosis inhibition and neuron protection of HBO can be increased through increasing the number of HBO treatment courses in neonatal rats with HIBD even if initiation of treatment is delayed after HIBD. PMID- 19558813 TI - [Effects of edaravone on IRE1 mRNA expression and neuronal apoptosis in the hippocampus of rats with status convulsivus]. AB - OBJECTIVE: To investigate the expression of the key marker of endoplasmic reticulum stress (ERS) IRE1 mRNA and neuronal apoptosis in the rat hippocampus after status convulsivus (SC), and the intervention effects of edaravone, a novel free radical scavenger. METHODS: Sprague-Dawley (SD) rats aged 19-21 days were randomly assigned to three groups: normal control, SC and edaravone-treated SC. SC was induced in the later two groups. The two groups were subdivided into 5 groups sacrificed at 4, 12, 24, 48, and 72 hrs after SC induction. IRE1 mRNA expression in the hippocampus was detected by RT-PCR. Neuronal apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL). The ultramicrostructural changes of neuron were observed by electron microscopy. RESULTS: IRE1 mRNA expression was obviously up-regulated 4 and 12 hrs after SC compared with the normal control group (P<0.01). IRE1 mRNA expression in the edaravone-treated SC group was notably higher than the untreated SC group 4, 12 and 24 hrs after SC and the normal control group (P<0.01). TUNEL positive cells in the hippocampus in the untreated SC group were significantly more than those in the normal control group (P<0.01). The number of TUNEL positive cells increased with the prolonged convulsion time. TUNEL positive cells in the edaravone-treated SC group were significantly reduced compared with those in the untreated SC group 12, 24, 48 and 72 hrs after SC (P<0.05 or P<0.01), but remained higher than the normal control group (P<0.05 or P<0.01). The peri-nucleus cell organ injuries were observed 4 hrs after SC and karyopycnosis and cytoplasm condensation were observed 12 hrs after SC in the SC and the edaravone-treated SC groups. The edaravone-treated SC group demonstrated less severe apoptosis than the untreated SC group. CONCLUSIONS: Edaravone may have neuroprotections against SC by an up regulation of IRE1 expression. It might serve as an effective agent for reducing ERS in vivo. PMID- 19558814 TI - [Effects of clearance of superoxide anion by catechin on the expression of NO and eNOS and apoptosis in endothelial progenitor cells induced by angiotensin II]. AB - OBJECTIVE: To evaluate the effect of clearance of superoxide anion by catechin on the expression of nitrogen monoxidum (NO) and endothelial nitricoxide synthase (eNOS) and apoptosis in endothelial progenitor cells (EPCs) induced by angiotensin II (Ang II). METHODS: The marrow endothelial progenitor cells of Sprague-Dawley rats were isolated and assigned to control (no treatment), Ang II treatment and Ang II + catechin treatment groups. After 48 hrs of culture, the concentration of O2*- in the supernate was measured by the NBT method, and NO concentration in the supernate was measured by the nitrate reductase method; the apoptosis rate of EPCs was detected by the TUNEL method; the mRNA expression of eNOS was detected by RT-PCR; the protein expression of eNOS was detected by Western blot analysis. RESULTS: Ang II of 10-6 mol/L was determined as the suitable concentration for cell induction by the MTT test. Catechin of 400 mg/L was determined as an advisable intervention dosage. The apoptosis rate of EPCs in the control, the Ang II and the Ang II+catechin treatment groups were 2.48+/ 0.12%, 54.18+/-0.77% and 16.87+/-0.35%, respectively, and there were significant differences among the three groups (P<0.01). The O2*- concentration in the Ang II and the Ang II+catechin treatment groups (81.7+/- 3.6 and 62.3+/- 2.2 U/L respectively) was significantly higher than that in the control group (33.7+/- 2.8 U/L) (P<0.01). An increased NO concentration was also found in the Ang II (189. 8+/- 9.0 micromol/L) and the Ang II+catechin treatment groups (276.4+/- 10.1 micromol/L) compared with that in the control group (105.8+/- 9.8 micromol/L) (P<0.01). There were significant differences in the concentrations of O2*- and NO between the Ang II and the Ang II+catechin treatment groups (P<0.05). The mRNA (P<0.05) and protein expression (P<0.01) of eNOS in the Ang II and the Ang II+catechin treatment groups increased significantly compared with those in the control group. The Ang II+catechin treatment group showed increased eNOS protein expression compared with the Ang II group (P<0.05). CONCLUSIONS: Ang II may induce the generation of O2*-, inactivate NO and increase gene and protein expression of eNOS in EPCs. Catechin might decrease the apoptosis of EPCs through the effective clearance of O2*-and the reduction of NO inactivation and of eNOS protein uncoupling. PMID- 19558815 TI - [Roles of enteric nervous system neurotransmitters and interstitial cells of Cajal in the colon in slow transit constipation in rats]. AB - OBJECTIVE: To evaluate the roles of enteric nervous system neurotransmitters, nitric oxide (NO), substance P (SP) and vasoactive intestinal polypeptide (VIP), and interstitial cells of Cajal (ICC) in the colon in slow transit constipation in rats. METHODS: Thirty-two healthy Wistar rats were randomly assigned to control and constipated groups. In the constipated group, the rats were daily administered with diphenoxylate (8 mg/kg) to develop slow transit constipation, while the control rats were fed with water. The number and the weight of fecal granule and the body weight of rats were recorded every 5 days for 90 days. Transit functions of intestinal movement were examined by an activated charcoal suspension pushing test one week after stopping the administration of diphenoxylate. The levels of NO and SP in the colonic mucosa were measured by nitrate reductase methods and ELISA respectively. The distribution of VIP and ICC positive cells confirmed with symbolic c-kit+ cells in the colonic wall were observed by immunohistochemical methods. RESULTS: The daily number of fecal granule in the constipated group was significantly less than that in the control group (P<0.01). The mean weight of each fecal granule in the constipated group was significantly higher than that in the control group (P<0.01). The discharge time of the first granule of black faeces in the constipated group (430.2+/- 132.1 min) was significantly longer than that in the control group (337.2+/- 74.7 min; P<0.05). There were no significant differences in NO and SP levels and the density of VIP positive cells in the distal colonic segment between the two groups. The number of c-kit+ cells in the distal colonic wall in the constipated group was significantly reduced compared with that in the control group (P<0.05). CONCLUSIONS: The reduction of ICC number in the distal colon may be contributed to the pathogenesis of slow transit constipation in rats. PMID- 19558816 TI - [Effects of DSCAM on differentiation of rat marrow mesenchymal stem cells into neurons in vitro]. AB - OBJECTIVE: To study the effects of Down syndrome cellular adhesion molecule (DSCAM) on differentiation of rat marrow mesenchymal stem cells (MSCs) into neurons in vitro. METHODS: MSCs from Sprague-Dawley rats were induced into neurons by baicalin. The expression of DSCAM before and after induction was evaluated by immunocytochemical staining and Western blot assay. After knockdown of DSCAM by siRNA transfection, the differentiation rate of neurons derived from MSCs was measured. RESULTS: Before induction, the expression of DSCAM was not detectable in MSCs. After bFGF preinduction for 24 hrs, DSCAM was slightly expressed in MSCs (1.71+/- 0.67%). The DSCAM expression increased 6 hrs after baicalin induction (15.79+/- 4.24%), reached a peak at 3 days (53.16+/- 5.94%) and then decreased gradually. The DSCAM expression 6 days after baicalin induction (28.99+/- 6.72%) was significantly lower than that at 3 days (P<0.01). However, after DSCAM-siRNA transfection, the DSCAM expression in MSCs was significantly reduced. MSCs did not express neuron-specific beta-III-tubulin before induction. After baicalin induction for 6 hrs, 3 days and 6 days, the expression of beta-III-tubulin was 1.40+/- 0.79%, 41.59+/- 3.17% and 59.11+/- 4.76% respectively. But the beta-III-tubulin expression significantly decreased 3 and 6 days after DSCAM-siRNA transfection (28.57+/- 2.91% and 43.90+/- 12.31% respectively). CONCLUSIONS: DSCAM may play an important role in MSCs differentiation into neural cells. PMID- 19558817 TI - [Comparison of immune function in children undergoing laparoscopic and conventional inguinal hernia repair]. PMID- 19558818 TI - [Serum CD4+ and CD8+ levels and the CD4+/CD8+ ratio in children with febrile convulsion]. PMID- 19558819 TI - [Therapeutic effect of ribavirin aerosol on herpangina in children]. PMID- 19558820 TI - [Misdiagnosis of appendicitis in children: analysis of 42 cases]. PMID- 19558821 TI - [One case report of congenital central hypoventilation syndrome]. PMID- 19558822 TI - [Sonographic finding in two children with testicular capillary hemangioma and review literature]. PMID- 19558824 TI - [Holoprosencephaly in a neonate]. PMID- 19558823 TI - [Severe enterovirus 71-associated hang-foot-and-mouth disease complicated by brainstem encephalitis: report of 2 cases]. PMID- 19558825 TI - [Research advance on Henoch-Schonlein purpura complicated by nervous system injury]. PMID- 19558828 TI - [Focusing on regulating surgical management of thyroid cancer]. PMID- 19558826 TI - [Research advance on placental iron transport proteins]. PMID- 19558829 TI - [Use of en bloc principle in re-operation of thyroid carcinoma]. AB - OBJECTIVE: The study is to explore the extent, main-point and use of en bloc principle in re-operation of thyroid carcinoma. METHODS: Clinical data of 75 patients received re-operation from 2002 to 2006 because of nonstandard operation were reviewed, including 10 men, 65 women, age ranging from 21-56. Ipsilateral completing lobectomy, isthmectomy, ipsilateral anterior cervical muscle, and level VI dissection were done in all the patients by en bloc principle. RESULTS: Residual tumor was found in 39 cases. There were, 10 in primary site, 8 in VI level, and 21 in both. Therefore tumor residual rate was 52.0%. The total metastatic lymph nodes in VI level were 63. Among the eight patients with recurrent laryngeal nerves paralyses which were hurt in the first operation, 4 were found been cut completely, 4 were ligated. The ligated locations were all near the place which the nerve enter the larynx. The ligated nerves were released, and the patients voice was improved greatly. CONCLUSIONS: Ipsilateral completing lobectomy, isthmectomy, ipsilateral anterior cervical muscle and VI level dissection is adequate for thyroid carcinoma patients who received nonstandard operation. The principle of en bloc resection can be used in the reoperation of thyroid carcinoma. PMID- 19558831 TI - [Diagnosis of thyroid space-occupying lesions using real-time contrast-enhanced ultrasonography with sulphur hexafluoride microbubbles]. AB - OBJECTIVE: To explore the value of contrast-enhanced ultrasound (CEUS) in the diagnosis of thyroid occupied lesions with injection of sulphur hexafluoride microbubbles. METHODS: Fifty nine cases of conventional ultrasonic diagnosis of thyroid lesions in patients with 73 lesions re-sulfur hexafluoride microbubble ultrasound contrast real-time inspection, the use of CEUS and contrast pulse sequencing (CPS). RESULTS: Seventy-three lesions were satisfied with the dynamic contrast perfusion imaging. Ultrasound contrast prompted the 15 lesions (13 cases) of malignant lesions by postoperative pathology confirmed papillary thyroid carcinoma, ultrasound images showed a low or mixed echo, boundary ambiguity, accompanied by Microcalcification foci, blood flow distribution I rank or grade III. Ultrasound prompted 58 lesions (46 cases) of benign lesions, blood flow distribution of grade II or III, the performance of a variety of two dimensional ultrasound image, showing cystic, solid or liquid-solid mass, border clearance. Contrast with the surrounding thyroid tissue, lesionsor=20 mm papillary thyroid carcinoma, manifested prior to the beginning of the adjacent thyroid substance to enhance and dissection, showing the form of high-enhanced perfusion; thyroid follicular adenoma as early as the beginning of the adjacent thyroid substance to enhance and dissection, mass retained within the contrast agent longer time, showing the form of high-enhanced perfusion; nodular goiter enhanced for thyroid nodules and adjacent synchronization started to pick up in real terms, after the 6 lesions showed nodular contrast slightly earlier than the adjacent thyroid substance started to pick up, 28 lesions with synchronous real beginning of the adjacent thyroid dissection, 11 lesions of nodular dissipated early on (started receding time < or = 25 s) adjacent to thyroid substance. CONCLUSION: The real-time CEUS with injecting sulphur hexafluoride microbubbles is valuable in the diagnosis and differential diagnosis of the thyroid occupied lesions. PMID- 19558830 TI - [Primary thyroid non-Hodgkin's lymphoma of Ann Arbor stages IE and IIE: prognostic factors]. AB - OBJECTIVE: To investigate the prognostic factors of primary non-Hodgkin's thyroid lymphoma. METHODS: From January 1981 to January 2008, 47 patients with stage IE and IIE pathologically confirmed as suffering from B cell non-Hodgkin's lymphoma and treated in hospital, were retrospectively analyzed. Pathology: diffuse large cell B-cell lymphoma (DLBCL) 28, mucosa-associated lymphoma (MALT) 19. The cancer specific survival (CSS) were calculated using the Kaplan-Meier method and compared by the Log-rank test. Age, sex, symptoms, extrathyroid extension, radiation doses, treatment modality and histological type characteristics were evaluated using the Cox regression analysis. RESULTS: The 5-year CSS rate was 61.8%. The 5-year CSS rate for the patients with primary tumor confined to thyroid and with extrathyroid extension were 86.7% and 50.0% (P=0.012). The 5 year CSS rate for the patients treated with radiation doses below 40 Gy and treated to 40 Gy or more were 30.8% and 79.0% (P=0.002). The 5-year CSS rate for the 18 patients with stage II received single modality therapy and the multimodality therapy were 33.3% and 61.1% (P=0.037). The 5-year CSS rate for patients with DLBCL lymphoma lesions and with MALT were 50.0% and 78.9% (P=0.038). Multivariate analysis by Cox regression showed that extrathyroid extension, radiation doses and histological type were independent prognostic factors. CONCLUSIONS: In primary non-Hodgkin's lymphoma of the thyroid, extrathyroid extension, radiation radiation doses and histological type are important prognostic factors. For patients with the stage II received multimodality therapy have a higher CSS than the ones received single-modality therapy. PMID- 19558832 TI - [Clinical features of parathyroid space-occupying lesions]. AB - OBJECTIVE: To investigate the clinical manifestation, diagnosis, treatment of parathyroid occupying lesions. METHODS: The clinical data of 42 patients with parathyroid occupying lesions were retrospectively analyzed, including the clinical symptoms and signs, laboratory results, pathologic and imaging results and treatment. RESULTS: The number of males and females were 8 and 34, with females: males ratio being 1:5.25. The median age was 39 years. There were 2 cases of parathyroid cancer, 29 cases of parathyroid adenoma, 11 cases of parathyroid cysts in this study. The symptoms were as follows: 40 cases of neck lump, 34 cases of osteoporosis/osteitis fibrosa cystica, 29 cases of urinary symptom, 7 cases of voice hoarseness, 4 cases of peptic ulcer, 3 cases of dyspnoea and dysphagia, 3 cases of thoracic cavity lump, 2 cases of enhanced amylase activity. Serum calcium ion level and serum parathyroid hormone (PTH) level were examined qualitatively before operation. Ultrasonography, ECT-99mTc MIBI, CT, MRI were used in diagnosing and locating parathyroid occupying lesions before operation. Twenty nine cases of parathyroid adenoma were treated with operation, 28 patients achieved complete remission, 1 suffered relapse after 23 months postoperative follow up. Eleven cases of parathyroid cysts were treated with operation and the outcome was no recurrence. Two cases of parathyroid cancer survived with out recurrence during follow up for 28 months and 50 months after operation. CONCLUSIONS: Examination of serum calcium and PTH level together with ultrasonography, ECT-99mTc-MIBI, CT, MRI is helpful to diagnose parathyroid occupying lesions. Surgery should be done as primary treatment. Tumor resection can be performed for parathyroid cysts, intraoperative exploration of bilateral neck is indicated for parathyroid adenoma, and a radical resection should be performed primarily for the parathyroid cancer. PMID- 19558833 TI - [Correlation between thyroid-stimulating hormone-peroxiredoxin1 signaling pathway and invasion of papillary thyroid carcinoma]. AB - OBJECTIVE: In order to assess the significance of thyroid-stimulating hormone receptor (TSHR), peroxiredoxin1 (Prx1), thioredoxin1 (Trx1), hypoxia inducible factor-1 (HIF-1)alpha in invasion and metastasis of papillary thyroid carcinoma (PTC). METHODS: The 34 paraffin-embedded specimen of classical PTC, 39 of tall cell variant of PTC and normal thyroid tissues adjacent to cancer operated in Tianjin Cancer Hospital from 2003 to 2007 were tested. The expressions of TSHR, Prx1, Trx1, HIF-1alpha in the specimen were assayed by immunohistochemical staining. The correlation between the 4 factors (TSHR, Prx1, Trx1, HIF-1alpha) and clinical features of PTC such as gender, age, tumor size, invasion, lymph node metastasis, histological type and T stage were studied. RESULTS: The rates of immuno-positive staining Prx1, Trx1 and HIF-1alpha in PTC were higher than the normal thyroid tissues adjacent to cancer with significant difference respectively (chi2 were 5.49, 6.16 and 40.48, P<0.05). The positive expression rate of TSHR in PTC was lower than the normal thyroid tissues adjacent to cancer (chi2=15.70, P<0.05), in tall cell variant of PTC was lower than classical PTC (chi2=4.24, P<0.05). The positive expression rates of Prx1, Trx1 and HIF-1alpha in PTC with local invasion, with lymph node metastasis, in tall cell variant and in stage T3-T4 were higher than PTC without local invasion, without metastasis, in classical variant and in stage T1-T2 with significant difference respectively (P<0.05). There were correlation between the expressions of Prx1 and Trx1, Trx1 and HIF-1alpha (r1 were 0.664 and 0.652, P<0.05). CONCLUSIONS: The high expressions of Prx1, Trx1 and HIF-1alpha were related to invasion and lymph node metastasis in PTC. TSH may promote the development of thyroid carcinoma by TSH Prx1-Trx1-HIF-1 signaling pathway. PMID- 19558834 TI - [Etiologic analysis of severe to profound hearing loss patients from Chifeng city in Inner Mongolia]. AB - OBJECTIVE: To investigate the etiology of patients with severe to profound hearing loss and to identify the ratio of hereditary hearing loss in Chifeng area in Northern China. METHODS: DNA were extracted from peripheral blood of 134 deaf patients from Chifeng special educational school and 100 normal hearing controls in Northern China. Audiology examinations showed that all patients had severe to profound bilateral sensorineural hearing impairment. Sequence analysis of the whole coding areas of GJB2, GJB3, GJB6, SLC26A4, mtDNA12SrRNA and mtDNAtRNASer(UCN) were performed. Individuals carrying SLC26A4 mutation were given further temporal bone CT scan. RESULTS: The ratio of hearing loss related to genetic factors in this population was 60.45% (81/134). About 33.58% (45/134) of the patients were given accurate genetic diagnosis. GJB2 mutations were responsible for approximately 17.16% of the cases in ChiFeng area. By screening SLC26A4 followed by temporal bone CT scan, we diagnosed 20 cases of enlarged vestibular aqueduct (EVA) and/or other inner ear malformation. SLC26A4 mutations account for about 14.93% of the cases. The aminoglycoside-related mtDNA 1555A>G mutation accounted for 0.76% of the cases in Chifeng area. In addition, another 13.43% (18/134) of the cases carried heterozygous GJB2 mutation and their hearing loss may be related to GJB2. 6.72% (9/134) of the cases carried heterozygous SLC26A4 mutation who were not found EVA by temporal bone CT or not took CT examination for some reasons. However, their hearing loss may also be SLC26A4 related. About 2.24% (3/134) of the cases carried mtDNA 12SrRNA 1095 T>C which may also be an aminoglycoside-related mutation and very likely be the cause of hearing loss. GJB3 might participate in the pathomechanism of hearing loss in 1.49% (2/134) of the patients. GJB6 mutation was not detected in this population. CONCLUSIONS: The ratio of hearing loss related to genetic factors in the sample drawing population from Chifeng was 60.45% (81 cases). GJB2 is the most common gene and SLC26A4 is the second common gene next to GJB2 that cause deafness in this area. PMID- 19558835 TI - [Preliminary study on intratympanic dexamethasone injection for management of patients with profound sudden hearing loss]. AB - OBJECTIVE: To evaluate the effects of intratympanic dexamethasone injection on patients with profound sudden hearing loss. METHODS: All patients in the present study were profound sudden hearing loss, with initial hearing loss of more than 90 dB, but no previous interventions was conducted on the 78 patients within 2 weeks onset of sudden hearing loss. Patients were assigned to 3 groups according to patients' choice, Group I (local and general dexamethasone administration, 22 cases), Group II (Intravenous dexamethasone injection, 44 cases), Group III (intratympanic dexamethasone injection, 12 cases). In addition, vessel dilation drugs, neurotrophic and hyperbaric oxygen therapy were also conducted on all patients. Intravenous dexamethasone was applied 15 mg/dx3 d, 10 mg/dx3 d and 5 mg/dx3 d, respectively. Intratympanic dexamethasone (5 mg/ml, 0.8 ml) injection was performed during 10 days (1 injection/2 days). Pure tone test was conducted on 10th, 20th, and 30th day after intervention. RESULTS: The factors which may impact on the prognosis were matched in all three groups. The threshold improvement more than 30 dB was 81.82% in group I, 83.3% in group II and 88.64% in group III. Statistical study showed there was no significant different among 3 groups (P=0.726). On the 30th day after intervention, pure tone threshold improvement was 41.36 dB in group I (local and general dexamethasone administration), 43.08 dB in group II (intravenous dexamethasone injection) and 51.70 dB in group III (intratympanic dexamethasone injection). Furthermore, pure tone threshold improvement among the 3 groups was no statistical different (F=1.58, P=0.2133). Obvious hearing improvement was noted on the 10th day after intervention, but no further improvement showed after 20 days intervention. More hearing improvement was revealed in the low frequency, while less hearing improvement was achieved in the high frequency. CONCLUSIONS: Comparison with intravenous dexamethasone injection, intratympanic dexamethasone injection did not provide more hearing improvement on patients with profound sudden hearing loss. PMID- 19558836 TI - [Measurement of exhaled nitric oxide in healthy Chinese]. AB - OBJECTIVE: To obtain the normal values of nitric oxide (NO) exhaled through nose and mouth in healthy Chinese adults by measuring exhaled NO and analyzing the influencing factors. METHODS: Eighty healthy Chinese adults were recruited, including 20 males and 60 females. The age ranged from 18 to 44 years old. Chemiluminescence analyzer (NIOX) was used to obtain the values of exhaled NO through nose and mouth. The relativity between NO and gender, age, height, body mass index, time, ambient NO were analyzed with Multiple linear regression and correlation. RESULTS: Exhaled NO values were (17+/-8)x10(-9) and correlated significantly with height. Regression equation: Y (exhaled nitric oxide)= 58.524+0.457X (height, cm), t=-2.985, P<0.01. Transnasal NO values were (819+/ 211)x10(-9) and correlated significantly with age and gender. Regression equation: Y (nasal nitric oxide)=760.245+9.417X1(age)-111.222X2(gender), t=5.188, P<0.01. CONCLUSIONS: Exhaled NO normal values were 17x10(-9) and Transnasal NO normal values were 819x10(-9). Exhaled NO correlated positively with height. Transnasal NO correlated positively with age and negatively with gender. PMID- 19558837 TI - [A huge true fibroma of nasal cavity in children: a case report]. PMID- 19558838 TI - [Analysis of clinical characteristics of chronic rhinosinusitis in patients with postirradiated nasopharyngeal carcinoma]. AB - OBJECTIVE: To study the characters of chronic rhinosinusitis in patients with irradiated nasopharyngeal carcinoma. METHODS: There were 65 cases of chronic rhinosinusitis after irradiated nasopharyngeal carcinoma (NPC, experimental group) and 65 cases of common chronic rhinosinusitis (CRS, control group) in the study. The visual analogue scale (VAS) was used to evaluate the intensity of subjective symptoms. Endoscopic finding was recorded and CT results were evaluated by Lund-Mackay scoring system. RESULTS: As to the VAS, nasal secretion was significantly more severe in experimental group (7.86+/-1.62), compared with control group (5.12+/-1.32, t=10.541, P<0.01). As to endoscopic finding, middle nasal meatus were clean in 35 (53.8%) cases in experimental group, and 23 cases (35.4%) in control group (chi2=4.483, P<0.05). CT score was (7.03+/-4.63) in experiment group, and (11.42+/-3.32) in control group (t=-6.207, P<0.05). The main reason lays in lower CT score and lower involved rate of ostiomeatal complex, frontal sinus, maxillary sinus, anterior ethmoid sinus. CONCLUSIONS: The characters of chronic rhinosinusitis in patients with irradiated nasopharyngeal carcinoma is quite different from the common CRS and different therapeutic measures should be taken. PMID- 19558839 TI - [Clinical study on 88 cases of adenoid cystic carcinoma in nasal cavity and paranasal sinuses]. AB - OBJECTIVE: To study the clinical characters, diagnosis, management and prognosis of patients with adenoid cystic carcinoma in nasal cavity and paranasal sinuses. METHODS: The data were analyzed retrospectively for 88 patients with adenoid cystic carcinoma in nasal cavity and paranasal sinuses, to evaluate the clinical characters, diagnostic and therapeutical aspects and the contribution of every factor influencing the survival. Survival analysis was performed by Kaplan-Meier method, comparison among/between groups was performed using Log-rank test, and multivariate analysis was carried out using Cox proportional hazard model. RESULTS: There were 56 patients in stage III and IV. Most of them received surgical operation combined with radiotherapy. The 5-year, 10-year and 15-year survival rates were 0.640, 0.341 and 0.190 respectively. While there were only 0.833, 0.221 and 0.323, 0.145 in stage III and stage IV lesions respectively. The 5-year and 10-year survival rates of 0.761, 0.415 were obtained in patients who received surgery combined with radiotherapy, 0.750, 0.367 and 0.286, 0.143 respectively in those treated by surgery and by radiotherapy alone. CONCLUSIONS: Advanced adenoid cystic carcinoma should be treated by combined surgical operation and radiotherapy. Stage and treatment approach are the independent factors affecting the prognoses of patients with adenoid cystic carcinoma in nasal cavity and paranasal sinuses. PMID- 19558840 TI - [Tracheoesophageal shunt for vocal rehabilitation after laryngectomy]. AB - OBJECTIVE: To discuss the clinical value of functional tracheoesophageal shunt for vocal rehabilitation after laryngectomy. METHODS: One hundred and twenty seven cases of tracheoesophageal shunt for vocal rehabilitation after laryngectomy in Cancer Hospital of Tianjin Medical University from 1981 to 2006 were analyzed retrospectively. RESULTS: Among 127 cases, 105 cases got successful phonation and the total success rate of vocal rehabilitation was 82.7%, all successful cases were followed up from 2 to 27 years had good phonation quality and no aspiration. Analyzing the reasons of failure in phonation of the 22 cases, 9 cases were because of improper operation (7 cases for narrow fistula and 2 cases for broad fistula), 13 cases were because of postoperative infection (10 cases for narrow fistula and 3 cases for broad fistula). The key to successful phonation was the size of fistula, the main causes of the failure in phonation were related to uncorrected operative procedure and postoperative infection. CONCLUSIONS: This method for vocal rehabilitation after laryngectomy has high success rate of vocal rehabilitation and low complications, it is relatively simple and worth popularizing in clinical treatment. PMID- 19558841 TI - [Three-dimensional anatomical research on sellar region for transnasal endoscopic surgery]. AB - OBJECTIVE: To develop a new method to explore the three-dimensional characteristics of anatomical structures at the sellar region in transnasal endoscopic surgery. METHODS: (1) The MicronTracker binocular visual navigation system was modified, and the tool's accuracy was tested by comparing the vernier caliper and turntable. (2) The basis nasi plane and median sagittal plane were used as datum plane, S point (the lateral margin point of nasal spine) and M point (maxillary line midpoint) were used as datum point to orientate the structures. The pitching angle, direction angle and distance of the important structures were measured by the tool designed by us based on the MicronTracker binocular visual navigation system and made a computer graphics model. RESULTS: (1) The tool's accuracy had no statistical difference as compared with vernier caliper and turntable. (2) The pitching angle, direction angle and distance of the important structures were obtained. (3) 3D-max 9.0 and AutoCAD-2008 were used to set up three-dimensional anatomical model of the anatomical structures. CONCLUSION: Based on the familiar point and datum of the skull, the orientation data of the important structures could be obtained and the three-dimensional model of the sella region anatomical structures could be constructed. PMID- 19558842 TI - [In vitro study of directional inducible differentiation of inner ear hair cells from human adipose-derived mesenchymal stem cells]. AB - OBJECTIVE: To explore the feasibility of directionally inducing human adipose derived mesenchymal stem cells (hAD-MSCs) towards inner ear hair cells. METHODS: Mesenchymal stem cells from human adipose tissue were isolated, purified and cultured in vitro. hAD-MSCs were induced to neural stem/progenitor-like cell, and then co-cultured with embryonic chick otic vesicle cells. Processed hAD-MSCs were tested by immunostaining to ascertain whether they expressed characteristic hair cell markers. RESULTS: Morphologically, hAD-MSCs were induced to differentiate into neural stem/progenitor cells and expressed specific neural markers. After being co-cultured with embryonic chick otic vesicle cells, hAD-MSCs expressed specific surface markers of inner ear hair cells. CONCLUSIONS: hAD-MSCs can be directionally induced to differentiate towards hair cell-like cells in vitro. PMID- 19558843 TI - [Influences of two different endolymphatic infusion ways to cochlear morphology and function in guinea pigs]. AB - OBJECTIVE: To explore the influence of two different endolymphatic infusion ways on cochlear morphology and function. METHODS: Forty healthy pigment guinea pigs (250 - 350 g) with normal Prey's reflex were divided into A and B group with 20 animals respectively. The right ears served as treated ears and the left ones as control ones. In group A, 5 microl of adenovirus 5-enhanced green fluorescence protein (Ad5-EGFP) suspension was infused into the scala media through an opened cochleostomy on the lateral wall of the scala media (LWS). In group B, the same volume of Ad5-EGFP suspension was infused into the scala media through punctured round window membrane and the basilar membrane (RBM). Cochlear morphology was examined under scan electric microscope and phalloidin staining was used to observe the hair cells in the infused ears after the animals were sacrificed. Auditory brainstem thresholds of the ears of all the animals were measured before and after treatment. RESULTS: All the animals recovered well after operation. The holes on the lateral wall of the scala media and punctures on the round window membrane were healed completely. EGFP labeling appeared in the organ of Corti and lining wall of the stria vascularis indicated that adenovirus suspension was injected into the scala media using LWS (succeed in 14 animals accounted for 70%) and RBM (in 8 animals accounted for 40%) ways. Viruses were inoculated into the scala media with only locally inflammation reaction. In group A the hearing threshold decreased significantly in the treated ears compared with the control ears after the operation [(33.1+/-10.3) dB, (9.4+/-3.9) dB, F=46.34, P=0.0005]. However, in group B there was no significantly different between the treated ears and the control ears after the operation [(2.5+/-3.8) dB, (2.5+/-3.8) dB, F=0.00, P=1.000]. Phalloidin staining indicated that in group A the extension of hair cells loss was bigger than in group B. In some animals of two groups, EGFP labeling appeared in the extra-lymphatic system indicating that some of the injected suspension leaked out of the scala media. CONCLUSIONS: Ad5-EGFP could be infused into the scala media through LWS or RBM and adenovirus could infect the lining cells of scala media and supporting cells in the basal membrane successfully without causing immunoreaction in the whole cochlea. LWS caused more hair cell loss and hearing loss than RBM. However, the cochlear morphology could be recovered completely after surgery. The positive inoculation rate was relatively higher that through LWS than that through RBM. PMID- 19558844 TI - [First visit for hoarseness: a rare case of a fish bone in paraglottic space]. PMID- 19558846 TI - [Diagnosis and surgery of primary tracheal neoplasms in children]. PMID- 19558845 TI - [Papillary thyroid carcinoma with tumor thrombus in internal jugular vein]. PMID- 19558847 TI - [A case report and literature review of branchio-oto-renal syndrome]. PMID- 19558848 TI - [Treatment for fibrous dysplasia invading interior orbital wall with nasal endoscopic surgery for two cases]. PMID- 19558849 TI - [Congenital arhinia: one case report]. PMID- 19558850 TI - [Right nonrecurrent inferior laryngeal nerve in a case]. PMID- 19558851 TI - [Bamboo nodes on the vocal cord and autoimmune disease]. PMID- 19558852 TI - [Diagnosis and management of thyroid carcinoma]. PMID- 19558854 TI - [Elicitation of mismatch negativity and its influence from stimuli deviant in the healthy youth]. AB - OBJECTIVE: To work out the elicitation plan, obtain the mismatch negativity (MMN) and get out the laboratory normal value as well as to study the influence to MMN from the deviation of auditory stimuli. METHODS: Hearing test of the tone burst stimulation was performed on 21 healthy young volunteers according to oddball stimulation sequence. Each subject was performed two kinds of auditory stimuli including frequency deviant stimuli and intensity deviant stimuli, and of each one included three series of stimulation. MMN was gained by subtracting the ERP of deviant stimuli from the ERP of standard stimuli. The latency and amplitude of each MMN were recorded, and then the effect of the deviant extent for MMN was analyzed. RESULTS: By this setup the MMN of normal young people was recorded and normal value of latency and amplitude of MMN were got. In the group of frequency deviant stimuli, the MMN latency [(155.81 +/- 29.08) ms], if the frequency was up to 2000 Hz, was shorter than that when the frequency deviance was 1000 Hz [(182.89 +/- 45.85) ms, (183.32 +/- 43.33) ms] (P = 0.033, 0.030); when the deviant extent were the same, the latency had no obvious difference if changing the frequency of the standard and deviant stimuli (P = 0.973); the MMN amplitude of three groups [(3.85 +/- 2.22) microV, (2.90 +/- 2.05) microV, (2.66 +/- 2.12) microV] had no obvious difference among them (P > 0.05). In the group of intensity deviant stimuli, the MMN latency [(157.04 +/- 34.87) ms], if the frequency was up to 20 dB, was shorter (P = 0.025, 0.017) than that when the intensity deviance was 10 dB [(184.46 +/- 38.05) ms, (186.24 +/- 42.36) ms]. When the deviant extent were the same, the latency had no obvious difference (P = 0.882) if changing the intensity of the standard and deviant stimuli but only group 4 and group 6 [(3.41 +/- 1.64) microV, (2.37 +/- 1.47) microV] were different in evidence (P = 0.031) while the others had no obvious difference (P = 0.524, 0.122). CONCLUSIONS: MMN was only related to the difference between standard stimuli and deviant stimuli, but there was no relationship between MMN and the notice, which indicate that MMN could objectively reflect the capability of brain to detect the change of stimuli. MMN is the representation of brain high level sensory function. PMID- 19558853 TI - [Audiological and genetic studies on 130 infants with hearing loss]. AB - OBJECTIVE: To investigate the genetic etiologies in the 0- 3-years-old infants with hearing loss and to analyze the interaction between genetics and environmental factors. METHODS: Total of 130 infants were performed detailed audiological evaluation as well as the detection of the popular deafness gene mutations in GJB2 gene, SLC26A4 and mtDNA12SrRNA. Of them, 84 cases were performed the computer tomography or magnetic resonance imaging examinations. RESULTS: Of the 130 cases, 54 infants were diagnosed as large vestibular aqueduct syndrome, while seven of 130 were as auditory neuropathy and the others were diagnosed as sensorineural hearing loss. Considering of the risks of etiologies for hearing loss, 85 of them had the experiences of the high risk factors at birth (65.4%, 85/130), while 23 of them had the exposure of aminoglycoside antibiotics, and 13 had the family history background as well as two cases were from the consanguineous families. In the causative genes screening, 42 infants were caused by the mutations of SLC26A4 gene (32.3%), but 14 infants found the mutations in GJB2 gene (4.6%), and no infants carried the mutation in mtDNA 12SrRNA 1555G and 1494T points in our studies. CONCLUSIONS: In our studies, about 36.9% infants hearing loss cases can be found the mutations in SLC26A4 and GJB2 genes. It is essential to put the idea into the hearing evaluation combined with genetic testing for the diagnoses of hearing loss. It is also helpful for exploring the etiologies of hearing loss and performing the target genetic consulting for decreasing the prevalence of deafness in the future. PMID- 19558855 TI - [Correlation between chirp auditory brainstem response and behavioral hearing threshold in children]. AB - OBJECTIVE: To investigate the correlation between thresholds in the chirp-ABR and behavior audiogram in order to find out if it is possible to be used as an clinical application of the chirp-ABR in estimating hearing sensitivity. METHODS: Twenty-two cases with hearing loss or normal hearing were enrolled in the study. The behavior audiogram and the response thresholds of chirp ABR (including chirp ABR, L-chirp ABR and U-chirp ABR) were obtained from 35 ears. RESULTS: Twenty-two cases were of both genders. The age was between 3.3- 6.5-years-old with the average age of 4.8-years-old. Divided by the degree of hearing loss, in the 35 ears, there were 6 with normal hearing, 2 with slightly hearing loss, 4 with moderate hearing loss, 10 with severe hearing loss and 13 with profound hearing loss. The Pearson correlation coefficients were 0.939, 0.900 and 0.930, respectively, which got from the data between the average of 0.5 - 4 kHz and chirp ABR respond threshold, 0.5 kHz and L-chirp ABR, and the average of 1 - 4 kHz and U-chirp ABR. CONCLUSION: As an objective test, the response threshold of chirp-ABR and the behavior audiogram were a highly correlated with each other, but more application in more subjects is needed. PMID- 19558856 TI - [Activation of nuclear factor-kappaB and aberrant gene expression of interleukin 6 in middle ear cholesteatoma]. AB - OBJECTIVE: To establish the activation of NF-kappaB in middle ear cholesteatoma, discuss the relationship of NF-kappaB and the gene expression of IL-6 and explore the pathogenesis of middle ear cholesteatoma. METHODS: Ten cases of cholesteatoma and 6 cases of normal external meatal skin were obtained from middle ear surgery. The NF-kappaB DNA binding activity and the mRNA level of IL-6 in these two kinds of tissues were detected by electrophoretic motility shift assay (EMSA) and Reverse transcription polymerase chain reaction (RT-PCR) respectively. The relationship of NF-kappaB DNA binding activity and the mRNA level of IL-6 were analyzed. RESULTS: The NF-kappaB DNA binding activities of cholesteatoma [(15.9 +/- 8.2)%] were higher than those in normal skin [(1.36 +/- 0.94)%, t = 3.502, P < 0.05]. The expression of IL-6 mRNA was increased significantly in patients with cholesteatoma, as compared with that in the control specimens (t = 2.166, P < 0.05) and had a significant positive correlation with NF-kappaB DNA binding activity (r = 0.752, P < 0.05). CONCLUSIONS: The IL-6 mRNA expression in cholesteatoma is closely related with the activity of NF-kappaB. It is tempting to speculate that NF-kappaB play a key role in the activation of cytokine in cholesteatoma. PMID- 19558857 TI - [Timing of operation and therapeutic effect of endoscopic optic nerve decompression for traumatic optic neuropathy]. AB - OBJECTIVE: To observe the optimal timing of operation and the therapeutic effect of endoscopic optic nerve decompression for traumatic optic neuropathy (TON). METHODS: The clinical records of 90 consecutive patients with TON (93 eyes) after head and/or maxillofacial trauma from April 1998 to March 2007 were reviewed and analyzed. All patients were either unresponsive or intolerant to medication before they underwent intranasal endoscopic optic nerve decompression. The time interval between the injury and operation ranged from one day to 97 days (median 5.5 days). Among the 93 eyes, there were 71 eyes with no visual acuity before operation and 22 eyes with residue visual acuity, including light perception in 1 eye, hand movement in 5 eyes, counting fingers in 13 eyes, 0.04 in 1 eye, and 0.1 in 2 eyes. Duration of follow-up ranged from 6 days to two years (median 8 days). RESULTS: After decompression, 35 patients (36/93 eyes, 38.7%) showed improvement of visual acuity, 53 patients (55 eyes, 59.1%) remained the same as before operation, while 2 patients (2 eyes, 2.2%) showed decreased visual acuity. Among patients with visual acuity beyond light perception before decompression, 68.2% of them (15/22 eyes) experienced visual improvement, whereas only 22.9% (8/35 eyes, 0.02 in two eyes) among patients who lost visual acuity immediately after injury, and 36.1% (13/36 eyes, 0.02 in five eyes) among those who lost visual acuity gradually after injury. There was a significant difference in visual improvement between group with visual acuity and group with no visual acuity (chi(2) = 11.864, P < 0.01). Among patients with no visual acuity, 41.2% of those (7/17 eyes) who underwent operation within 3 days of injury, experienced improvement in visual acuity, compared with 25.9% (14/54 eyes) for those who underwent the operation more than 3 days after injury. It was indicated that no significant difference in visual improvement between these two groups (chi(2) = 1.46, P > 0.05). When comparing different sites of fracture, the effect of surgery was the most desirable (55.6%, 10/18 eyes improved) if the fracture occurred simultaneously in both exterior and interior walls of optic canal, followed by the interior wall fracture (45.7%, 21/46 eyes). The operation was less effective if there was no fraction (20%, 4/20 eyes) or if the fracture occurred in exterior wall alone (11.1%, 1/9 eyes). CONCLUSIONS: Endoscopic optic nerve decompression is a minimally invasive procedure with no adverse cosmetic effects. Early operation is recommended for saving vision, even though visual acuity is lost immediately after injury. However, the satisfactory clinical effects of endoscopic optic nerve decompression require further study. PMID- 19558858 TI - [Roles of epidermal growth factor receptor signaling pathway on cultured human nasal epithelial cells RPMI-2650]. AB - OBJECTIVE: To explore the roles of epidermal growth factor receptor (EGFR) signaling pathway on cultured human nasal epithelial cells RPMI-2650. METHODS: RPMI-2650 cells were cultured in vitro, the growth curve was measured and the ultrastructure was observed using scanning electron microscope. When the cells were significantly confluent, they were divided into 4 groups, group A: maintained in Eagle's minimum essential media (EMEM) medium without adding any stimulators; group B: added with epidermal growth factor (EGF) 25 ng/ml; group C: added with AG1478 (EGFR selective inhibitor) 10 micromol/L followed by EGF 25 ng/ml 30 minutes later; group D: added with PD98069 (p44/42MAPK selective inhibitor) 30 micromol/L followed by EGF 25 ng/ml 30 minutes later. After incubated for 24 hours, the expression of EGFR and MUC5AC proteins in the cells of these 4 groups was studied using cytoimmunity and Western blotting. RESULTS: RPMI-2650 cells were significantly confluent after incubated for 5 to 7 days. The shape of cells was round or oval, and a large number of microvilli covered to their surface but without cilia under scanning electron microscope. The EGFR protein was expressed in the cells of group A and D, abundantly in group B, while weakly in group C. The values of comparative absorbance had significant difference between group A, B, D and group C, respectively (P < 0.01). For the MUC5AC protein, its expression was strong in the cells of group A, abundant in group B, and weak in group C and D. Significant difference of the values of comparative absorbance was analyzed between group B and group C, D, respectively (P < 0.01), while no difference between group C and group D (P > 0.05). CONCLUSIONS: The production of MUC5AC in human nasal epithelial cells RPMI-2650 is regulated via the expression and activation of epidermal growth factor receptor signaling pathway. PMID- 19558859 TI - [Relationship between nasal discharge eosinophils and responses to treatment of inhaled glucocorticosteroid in patients with allergic rhinitis]. AB - OBJECTIVE: To evaluate the treatment responses of persistent allergic rhinitis with and without nasal discharge eosinophilia (EOS) to inhaled glucocorticosteroid (CS), and therefore to verify whether low nasal discharge eosinophils predict poor response to treatment with CS. METHODS: Forty-two symptomatic allergic rhinitis patients, who had not received CS therapy in three months preceding the study, were examined before and 2 month,4 months and 6 months after treatment with CS. At each visit, all patients underwent symptom scoring and physical sign scoring. The level of eosinophil cationic protein (ECP) in the nasal discharge supernatants was measured by radioimmunoassay. The patients were divided into 2 groups according to nasal discharge EOS percentages, an EOS group (group A, EOS > or = 0.03) and a non-EOS group (group B, EOS < 0.03). The response to CS therapy (as measured by symptom and physical sign scores) and the changes of nasal discharge measurements were compared between the 2 groups. RESULTS: In the group A, the baseline EOS [0.086 (0.065; 0.176)] and ECP level [(326 +/- 145) microg/L] were significantly higher than those of the group B [0.016 (0.005; 0.022)] and ECP level (154 +/- 58) microg/L], respectively, t = 4.40, 3.33, both, all P < 0.01. After 2 month and 6 months CS therapy, the nasal discharge EOS, ECP pred were 0.038 (0.006; 0.070), 0.019 (0.010; 0.060), (175 +/- 122) microg/L, (175 +/- 153) microg/L, respectively in the EOS group, which were significantly different as compared to baseline values (F = 6.73, 7.38, respectively, all P < 0.05). But in the non-EOS group, the nasal discharge EOS ECP pred were 0.014 (0.004; 0.032), 0.015 (0.000; 0.026), (118 +/- 60) microg/L, (112 +/- 60) microg/L, respectively at 2 and 6 months, which showed that the the nasal discharge EOS pred and the symptom and physical sign scores improved did not change (F = 0.82, P > 0.05), but the ECP level improved (F = 3.78, P < 0.05). and the average daily dose of CS wear not different between the two groups at any visits. CONCLUSIONS: In persistent allergic rhinitis with low nasal discharge EOS, CS therapy for 6 months failed to improve symptom and physical sign. PMID- 19558860 TI - [Value of pulse transit time in detecting respiratory drive]. AB - OBJECTIVE: To assess the accuracy of pulse transit time (PTT) in classification of apnea events, and collect data for clinical application reference. METHODS: Thirty-two obstructive sleep apnea-hypopnea syndrome (OSAHS) patients included in the research had Polysomnography (PSG), and 10 305 apnea events were recorded. All the events were analyzed by PTT and esophageal pressure (Pes) respectively. The results were analyzed to assess the accuracy of PTT and compare the accuracy of pulse transit time between REM stage and NREM stage, and analyze the correlation between age, body mass index (BMI), apnea hypopnea index (AHI) and concordance rate in every patient. RESULTS: The total concordance rate between PTT and Pes in classification of apnea was 96.7% (9970/10305). The sensitivities of PTT in detecting central, mixed and obstructive apnea were 88.0%, 91.3% and 97.8% respectively and the specificities were 99.8%, 97.8% and 92.8% respectively. The false determinations of apnea events mainly concentrated on the false determinations between the obstructive and mixed apnea. There was no statistical significant between the accuracy of PTT in different sleep stages. There was a negative relationship between the age, BMI, Lowest SaO2, AHI and the concordance rate. CONCLUSIONS: There was good concordance between PTT and Pes in classification of apnea. PTT had very high sensitivity and specificity in detecting all kinds of apnea. This study showed that PTT can detect respiratory drive noninvasively with high accuracy. PMID- 19558861 TI - [Etiology of inspiratory laryngeal stridor in children]. AB - OBJECTIVE: Retrospective analysis was performed on the etiology of inspiratory laryngeal stridor in children. The purpose is to raise the diagnosis and cure rate of the disease. METHODS: All patients were hospitalized in Children's Hospital from Jan, 2005 to Jan, 2007. Among of them, 245 cases were male and 133 cases were female. The median age was 4 months (range from 12 hours to 30 months). All the patients had chest X-ray examination. Two hundred and eighteen cases received chest CT scan, video laryngoscope, direct laryngoscope and bronchofibroscopy. RESULTS: The diagnosis were as follows: acute laryngitis (140 cases), laryngomalacia (117 cases), acute laryngotracheal bronchitis (54 cases), vocal cord paralysis (18 cases), congenital tracheomalacia (9 cases), congenital laryngeal webs (8 cases), congenital cleft of larynx (6 cases), laryngeal cyst (6 cases), laryngeal papilloma (6 cases), acute epiglottitis (4 cases), congenital infraglottic stenosis (3 cases), tracheobronchial foreign body (3 cases), cysts thyrolinguals (1 case). All cases were cured except congenital tracheomalacia (9 cases), congenital cleft of larynx (6 cases), laryngeal papilloma (6 cases), congenital infraglottic (3 cases). CONCLUSIONS: The etiology of inspiratory laryngeal stridor in children are very complicated. Video laryngoscope is recommended for all cases except for the acute inflammation disease. Chest CT scan and bronchofibroscopy may be necessary for some cases. PMID- 19558862 TI - [Clinical analysis on giant lymph node hyperplasia on neck]. AB - OBJECTIVE: To improve the diagnosis and management level of giant lymph node hyperplasia (Castleman's disease). METHODS: To retrospective analyze 10 misdiagnosed cases with Castleman's disease in order to give some suggestions for clinical diagnosis and differential diagnosis. RESULTS: Ten patients with neck giant lymph node hyperplasia underwent surgical treatment after misdiagnosis. There were 8 localized Castleman's disease constituted of 6 cases with hyaline vascular type and 2 cases with mixture type and 2 multicentric Castleman's disease constituted of 1 cases with plasma cell type and 1 cases with mixture type were classified according to the criteria described by Frizzera. Ten cases were diagnosed by secondary operation after misdiagnosis and were clinically characterized by painless neck lymphadenectasis, 2 cases with multicentric Castleman's disease accompanied with aspecific systemic symptom and (or) multi system damage. Ten cases survived for 4 - 17 years during follow-up periods in which 1 case with plasma cell type, multicentric Castleman's disease was recurrent 2 years later and underwent lymphadenectomy and chemotherapy and have no local recurrence so far. CONCLUSIONS: Castleman's disease on neck is seldom seen and liable to misdiagnose. The diagnosis of Castleman's disease is based on its histopathological characteristics by lymph node resection biopsy. It should be considered in the differential diagnosis with lymph node tuberculosis, lymphadenitis, sarcoidosis and granuloma. Operation is the first choice for patient with localized type and multicentric type without serious involvement of multiple system functions. PMID- 19558863 TI - [Cyclin D1 gene G870A polymorphism and susceptibility to nasopharyngeal carcinoma]. AB - OBJECTIVE: The aim of this study was to investigate the susceptibility and prognostic implications of the cyclin D1 gene (CCND1) G870A polymorphism to nasopharyngeal carcinomas (NPC) in Han population in Yunnan China. METHODS: Two hundred and forty one cases with NPC and 271 matched cancer-free controls were genotyped for the CCND1 G870A polymorphism by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and sequencing. The adjusted odds ratios (OR) and 95% confidence intervals (CI) were calculated by using unconditional logistic regression model. Overall survival was assessed using univariate and multivariate analyses. RESULTS: Contrast with homozygous CCND1 G870G, A allele significantly increasing risk of NPC was associated with homozygous A870A (OR = 4.79, 95% CI 2.77 - 8.28, P < 0.001) and heterozygous A870G (OR = 1.72, 95% CI 1.10 - 2.68, P = 0.017). The subjects at least having one CCND1 870A allele had OR of 2.40 (95% CI 1.59 - 3.63, P < 0.001). Furthermore, smoking may increase the risk of developing NPC interacting with CCND1 G870A polymorphism. Kaplan-Meier analysis and Cox regression analysis demonstrated that the five-year survival rate of subjects with AA, AG and GG genotype was 56.2%, 78.5% and 81.4% (AA vs GG, P = 0.003; AA vs AG, P = 0.012; AG vs GG, P = 0.132), but not independent prognostic factor in NPC (P = 0.501). CONCLUSIONS: The CCND1 870A allele is associated with the NPC in Han population in Yunnan China, meanwhile, showed a significant prognosis for those patients. PMID- 19558864 TI - [Clinicopathological features and prognosis of nasopharyngeal adenocarcinoma]. AB - OBJECTIVE: To investigate the clinical manifestations, EB virus serology and treatment outcome of nasopharyngeal adenocarcinoma (NPAC). METHODS: Clinical records of NPAC patients between 1964 and 2000 in Cancer Center of Sun Yat-sen University were retrospectively reviewed. RESULTS: Among 48 patients with NPAC, 45.2% (7 cases of N1, 8 cases of N2 and 4 cases of N3) of them presented with cervical metastasis. Pathologically, common type and salivary gland type of NPAC accounted for 58.3% (28 cases) and 41.7% (20 cases) respectively. The positive rate of the EB virus antibody VCA-IgA was 56.7% in the whole group and only 23.7% in the salivary gland type of NPAC. The overall local control rate and the 5-year disease free survival rate by Kaplan-Meier method were 87.0% (40/46) and 65.2% respectively. Baseline data analysis showed that age, gender, N stage and M stage were not the significant factors, never the less the T stage was not balanced between the two groups (surgery plus radiotherapy vs radiotherapy alone, chi2 = 4.801, P = 0.045). The patients treated by surgery plus radiotherapy had significantly higher 5-year disease free survival rate than by radiotherapy alone (88.9% vs 74.7%, Log Rank test: chi2 = 4.272, P = 0.039). Cox's multivariate analysis showed treatment modality and N stage were the significant factors influencing survival (RR were 15.276 and 6.529, P < 0.05). CONCLUSIONS: NPAC is a distinct entity in all types of nasopharyngeal carcinoma. EB virus serology has limited value in its diagnosis. Surgery plus radiotherapy could be another choice of treatment for early lesions of NPAC. PMID- 19558865 TI - [Distribution of dexamethasone in cochlea after intratympanic injection in rats]. AB - OBJECTIVE: To investigate the distribution and pharmacokinetics of dexamethasone of different concentrations in the inner ears of SD rats after intratympanic injection. METHODS: Totally 144 adult SD rats were anaesthetized and dexamethasone sodium phosphate of different concentrations (5 mg/ml, 10 mg/ml, 20 mg/ml) was injected into the tympanums. The rats were sacrificed at various postinjection survival times (5 min, 10 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 48 h, 72 h), and every 4 rats were included into each group. Then after a series of processes the inner ear tissue was cryostat sectioned. The distribution of dexamethasone was evaluated using immunofluorescence with semiquantitative analysis. Immunofluorescence was also used in another 4 normal SD rats to detect the distribution of Glucocorticoid receptor (GR) in the inner ear. RESULTS: Dexamethasone was observed initially 15 min after local drug administration and 30 min to its peak level. The highest concentration of dexamethasone labeling was seen in the spiral ligament, organ of Corti and spiral ganglion, which paralleled the distribution of GR. The tissue concentration of 10 mg/ml and 20 mg/ml groups was higher than 5 mg/ml every corresponding time point, and the lasting time was also prolonged from 48 hours to 72 hours. CONCLUSIONS: Dexamethasone can enter into the cochlear tissue quickly after transtympanic injection, and its distribution accords nearly exactly with that of GR. Increase of the concentration of dexamethasone results in higher tissue distribution and longer lasting time. PMID- 19558866 TI - [Effects of glucocorticoid on calcium-activated chloride channel expression in nasal mucosa in allergic rhinitis rats]. AB - OBJECTIVE: To investigate the expression of CLCA3 and Muc5ac in nasal mucosa in allergic rhinitis rats and the effects of glucocorticoid on its expression. METHODS: Thirty SD rats were randomly divided into allergic rhinitis group, dexamethasone group and control group. Expression of CLCA3 mRNA and Muc5ac protein in nasal mucosa were detected by RT-PCR and immunohistochemical assay, respectively. RESULTS: CLCA3 mRNA and Muc5ac protein in allergic rhinitis group were significantly higher than those in control group (t = 8.565, 5.317, P < 0.01, respectively). The increased expression of CLCA3 mRNA in allergic rhinitis group was well correlated with the expression of Muc5ac protein and the correlation coefficient was 0.813 (P < 0.05). After treatment with dexamethasone, the expression of CLCA3 mRNA and Muc5ac protein was notably lower than that in allergic rhinitis group (t = 3.102, 2.226, P < 0.05, respectively). CONCLUSIONS: The stronger gene expression of CLCA3 exists, complicated with mucus overproduction in the nasal mucosa of allergic rhinitis rats. CLCA3 expression may play a pivotal role in mucus overproduction in allergic rhinitis. Dexamethasone substantially downregulates the expression of CLCA3 mRNA and Muc5ac protein. PMID- 19558867 TI - [Establishment and pathophysiological observation of an experimental animal model of traumatic optic neuropathy]. AB - OBJECTIVE: To establish an animal model of traumatic optic neuropathy (TON) that resemble to clinical state and study the mechanical principle and change of pathophysiology of its nerve injury for clinical diagnosis and treatment. METHODS: New Zealand white rabbits were used as the research objects. The method introduced by Wang Yi was repeated and improved. Mild and severe animal models of TON were established by reformed Wang Yi operation separately. After the spring gun struck, all animals were observed on pupils and direct light reflex and received the examinations of pattern reversal visual evoked potentials (PR-VEP). The pathophysiology of normal and injury optic nerve was observed. RESULTS: After recovery from anesthesia, the mydriasis and disappearance or dullness of direct light reflex happened in all injured eyes. No brain contusion, infection, orbital fracture and death were found. One optic nerve was broken with complete tunica vaginalis. The latency and amplitude of injured eyes deteriorated gradually. In group B, the waves became flat rapidly. After injury, the optic nerve underwent 3 stages: edema, hyperplasia and atrophy. The pathomorphological changes of injured eyes in group B were more serious than that in group A in any time. CONCLUSIONS: The reformed operation can establish constant nerve injury with high success rate. In mildly injured eyes, the injury deteriorated gradually. However, part visual function remained. In severely injured eyes, the pathomorphological changes were irreversible sooner after struck, and the visual function lost completely. There is a good correlation between PR-VEP and pathomorphology. PR VEP can guide the clinical diagnosis and treatment. PMID- 19558869 TI - [Mucociliary ultrastructure and distribution of maxillary natural ostium]. PMID- 19558868 TI - [Establishment of the rabbit VX2 tumor as a model for pyriform sinus carcinoma and observation of its biological features]. AB - OBJECTIVE: To establish the rabbit VX2 tumor as a model for pyriform sinus carcinoma and to observe its biological features. METHODS: VX2 tumor was implanted into the pyriform sinus of 15 rabbits by direct laryngoscope. Fifteen rabbits were randomized into 3 groups (average of 5 rabbits per group). Observation of the tumor growth and evaluation of the histopathological characterization were taken on one group each at the of time 14, 21 and 28 days after transplantation respectively. RESULTS: Tumors were found grown in the pyriform sinus of all 15 rabbits with a success implantation rate of 100%. Dysphagia, body weight loss, rhinorrhea and short of breath appeared in the rabbits 28 days after transplantation. The metastasis rates of deep cervical were 100% in all three groups. The metastasis rates of submandibular lymph nodes were 3/5, 4/5 and 5/5 in 14-day, 21-day and 28-day group respectively. The metastasis rates of paratracheal lymph nodes were 0, 4/5 and 5/5 in 14-day, 21-day and 28 day group respectively. There were opposite side lymph node metastasis of deep cervical, submandibular and paratracheal in 4, 3 and 5 rabbits on 14, 21 and 28 days after transplantation respectively. The median diameter for deep cervical, submandibular and paratracheal neck lymph nodes were 1.50, 0.60 and 0.0 cm in 14 days; 1.60, 0.80 and 0.50 cm in 21 days; 1.80, 0.8 and 0.65 cm in 28 days (P > 0.05). CONCLUSIONS: The animal model for pyriform sinus carcinoma is established successfully. The metastasis of deep cervical lymph node could be induced from day 14 after VX2 transplantation. PMID- 19558870 TI - [A case of metallic foreign body pushed away from eustachian tube by electronic nasopharyngoscopy and flexible forceps]. PMID- 19558872 TI - [A case of adenoid cystic carcinoma after tympanoplasty]. PMID- 19558871 TI - [One case of huge facial neurilemmoma in parotid region]. PMID- 19558873 TI - [Summary of 2008 national conference on diagnosis and non-surgical treatment of otological diseases]. PMID- 19558874 TI - [Diagnosis and treatment of syndrome of styloid process]. PMID- 19558875 TI - [Central issues in the development of laryngology]. PMID- 19558877 TI - [Guideline for the diagnosis and surgical treatment of obstructive sleep apnea hypopnea syndrome]. PMID- 19558876 TI - [Explanation of guideline for the diagnosis and surgical treatment of obstructive sleep apnea hypopnea syndrome]. PMID- 19558878 TI - [Summary of 2008 national conference on laryngological and tracheal diseases]. PMID- 19558879 TI - [Pepsin immunoassay in the sputum for detection of laryngopharyngeal reflux]. AB - OBJECTIVE: To study whether the presence of gastric pepsin in the sputum might be used as a reliable criteria of laryngopharyngeal reflux. METHODS: Fifty-six patients with the symptoms of laryngopharyngitis and fifteen healthy people were recruited. Fifty-six patients were divided into laryngopharyngeal reflux group and chronic laryngitis group by the reflux symptom index (RSI), by the reflux finding score (RFS) and by their treating experiment taking omeprazole 20 mg bid for 2 weeks. Sputum in all three groups was obtained in the morning. Pepsin in the sputum was measured by enzyme-linked immunoadsorbent assay. RESULTS: The positive rate of pepsin in sputum among LPR group, chronic laryngopharyngitis group and normal group were 93.8% (30/32), 75.0% (18/24), 20.0% (3/15) respectively, and the median concentration of pepsin were 5.3 [1.3; 53.4] ng/ml, 0.8 [0.1; 17.2] ng/ml, 0.0[0.0;0.0] ng/ml (H = 23.29, P = 0.000). Compared with co-diagnosis as gold standard, the sensitivity of RSI, RFS treating experiment and the pepsin immunoassay was 59.4%, 84.4%, 81.3% and 93.8%, and the specificity of those was 87.2%, 61.5%, 95.8% and 46.2% respectively. CONCLUSIONS: Detection of pepsin in sputum by immunoassay might provide a high sensitive, noninvasive method for laryngopharyngeal reflux. PMID- 19558880 TI - [One case of death due to glossopharyngeal nerve microwave-coagulation in treatment of refractory glossopharyngeal neuralgia]. PMID- 19558881 TI - [A pilot study on the diagnosis of laryngopharyngeal reflux diseases]. AB - OBJECTIVE: To study the value of the reflux symptom index (RSI) and reflux finding score (RFS) in laryngopharyngeal reflux disease (LPRD). METHODS: Twenty five patients with RSI scores > 13 or RFS scores > 7 and sixteen patients with RSI scores > 13 and RFS scores > 7 were suspected as LPRD. Forty one patients with the RSI scores > 13 and/or RFS scores > 7 were suspected as LPRD, 15 patients with the RSI scores 13 and RFS scores > 7 (Kappa = 0.55, u = 3.06, P < 0.01). There was a significant decreasing in RSI and RFS scoring in patients after anti-acid treatment for 3 months (t(RSI) = 8.838, P(RSI) = 0.000; t(RFS) = 5.695, P(RFS) = 0.000). CONCLUSIONS: The RSI and RFS can be used as screening tool for laryngopharyngeal reflux diseases and as a simple method for evaluating the effectiveness of treatment. PMID- 19558882 TI - [Self-assessment characteristics of voice handicap index for voice disorders and its influencing factors]. AB - OBJECTIVE: To investigate the self-assessment characteristics of Voice Handicap Index (VHI) for voice disorders and its influencing factors. METHODS: One thousand seven hundred and sixty six dysphonic patients and 120 control subjects were included in this study. Two hundred twenty seven patients were treated with phonosurgery or Botulinumtoxin injection. VHI was used for self-assessment. RESULTS: Dysphonic patients had worse VHI scores than control (z from 8.039 to 17.043, P = 0.000). There was a significant difference among the VHI scores of different diseases. VHI scores were descending in order among spasmodic dysphonia, vocal fold paralysis, functional dysphonia, sulcus vocalis, benign and malignant tumor of vocal fold, vocal fold cyst, Reinke's edema, vocal fold polyp, vocal fold keratosis and chronic laryngitis, vocal nodule. The emotional scores were the highest in spasmodic dysphonia, and followed by functional dysphonia. In another group, the physical scores were higher than functional scores and emotional scores. Treatment resulted in statistical improvement in VHI scores (P < 0.05). The total scores were different significantly between different educational background and age groups (F from 8.701 to 27.371, P = 0.000). The higher the educational degree, the higher the VHI scores. As to age groups, the juvenile group's scores were the lowest, while the youth's group the highest, then the scores declined when ages increased. CONCLUSION: As a useful supplementary instrument to measure the voice disorder severity and the treatment's effect, VHI can comprehensively assess the voice handicap's affect to the life quality and the difference after the treatment, especially in physical, functional and emotional aspects, but it is somehow subject to the educational degree and age. PMID- 19558883 TI - [Experience of diagnosis and treatment for cervical tracheoesophageal fistula complicated with laryngotracheal stenosis]. AB - OBJECTIVE: To explore the treatment of cervical tracheoesophageal fistula (TEF) with complicated or remnant laryngotracheal stenosis (LTS) and anterior neck defect (AND). METHODS: From 1980 to 2007, 14 patients were diagnosed as TEF. Among them, 9 patients had complicated or remnant LTS, 3 patients had complicated AND, and 2 patients had TEF which were induced by Nickel-Titanium alloy mesh stent for treating benign esophageal stricture. All these patients were retrospectively studied in Tangdu Hospital. Treatment consisted of conservative therapy of TEF, staged surgical repair of TEF and laryngotracheal reconstruction according to the dimension (small or large) of TEF and complications. RESULTS: Four patients with small TEF (2 - 3 mm length) complicated LTS underwent laryngotracheal reconstruction stented with silicone T tube and TEF was adopted conservative treatment. The TEF and LTS were healed. Six patients with larger TEF (10 - 25 mm length) were repaired by staged surgical repair of TEF and laryngotracheal reconstruction. Among them, 3 cases had complicated LTS and AND, 2 cases had recent LTS and 1 case had TEF without complication. Two patients had TEF and LTS, whose TEF healed before laryngotracheal reconstruction, the remnant LTS were reconstructed and healed. During the follow-up ranged from one to ten years, 12 patients were successfully treated without complications. One patient with TEF and LTS was treated only LTS because of a segment of esophagus was closed and treated with esophagogastrostomy in the department of thoracic surgery after LTS was successfully reconstructed and cured. One patient died of bleeding and asphyxia induced by the Nickel-Titanium alloy stent because of the stent had not been taken out. CONCLUSION: The small cervical TEF complicated or remnant LTS can be treated by laryngotracheal reconstruction and conservative treatment of TEF at the same time. A larger TEF complicated LTS should be treated by staged repair of TEF and LTS. PMID- 19558884 TI - [Congenital stenosis of external auditory canal with cholesteatoma and skin fistulae or sinuses]. AB - OBJECTIVE: To investigate the clinical features, differential diagnosis and management of congenital stenosis of external auditory canal (CSEAC) with cholesteatoma. METHODS: The clinical information for 10 cases of CSEAC with cholesteatoma was retrospectively reviewed. RESULTS: The patients' ages ranged from 4.75 to 22 years (average 12 years). The diameter of the external auditory canal (EAC) was < 2 mm. All 10 ears had a history of postural fistulae or sinuses. Bone erosion of EAC was distinctly shown in high-resolution computed tomography (HRCT) of all cases, as well as soft tissue masses, which led to enlargement of the bony canals. All patients underwent canaloplasty; eight ears received hearing reconstructions at the same time. Cholesteatoma in EACs was confirmed during the operations, accompanied by compression and destruction of the post-superior and/or inferior bony wall. Postoperative pathologic examinations proved the diagnosis of cholesteatoma, and excluded any tissue of bronchial cleft cyst or fistula. After a follow-up 1 to 3 years, no recurrent cholesteatoma was found in any of the 10 cases. All reconstructed EACs were clean and smooth. The hearing levels in the eight ears that received hearing reconstructions improved 20 - 35 dBHL. CONCLUSIONS: In CSEAC with cholesteatoma, the bony wall of EAC is most commonly involved. This involvement will lead to bone erosion of the EAC and may subsequently lead to the formation of postural or cervical sinuses. HRCT of temporal bone can show characteristic signs of soft tissue mass in EAC, with adjacent bone erosion. PMID- 19558885 TI - [Correlation between four-phase rhinomanometry and acoustic rhinometry]. AB - OBJECTIVE: To obtain the normal values of four-phase rhinomanometry specific parameters of normal adult Chinese and analyze the correlation between four-phase rhinomanometry and acoustic rhinometry measurement results. METHODS: Eighty-five normal adults were recruited. The HRR2 four-phase rhinomanometry was used to acquire the effective resistances in inspiration, expiration and total breathing process (Reffin, Reffex, Refft) and vertex resistance in the process of inspiration and expiration (Vrin and Vrex). The Eccovision acoustic rhinometry was used to measure the minimum cross-sectional area (MCA) and the nasal volume of 0-5 cm nasal cavity (V5). RESULTS: Reffin (x(-) +/- s) was (1.28 +/- 1.02) Pa/(cm(3).s) for male, (1.55 +/- 1.03) Pa/(cm(3).s) for female; Reflex (x(-) +/- s) was (1.43 +/- 1.07) Pa/(cm(3).s) for male, (1.75 +/- 1.14) Pa/(cm(3).s) for female; Refft (x(-) +/- s) was (1.34 +/- 0.99) Pa/(cm(3).s) for male, (1.62 +/- 1.03) Pa/(cm(3).s) for female; Vrin (x(-) +/- s) was (1.31 +/- 1.03) Pa/(cm(3).s) for male; (1.60 +/- 1.03) Pa/(cm(3).s) for female, Vrex (x(-) +/- s) was (1.46 +/ 1.04) Pa/(cm(3).s) for male, (1.82 +/- 1.17) Pa/(cm(3).s) for female. No statistically significant difference was found between men and women (r = 0.661, 0.397, 0.127, 0.649, -0.684, P > 0.05, respectively). There was no significant correlation between Reffin, Reflex, Refft, Vrin, Vrex and age, height, weight, head circumference, body surface area, body mass index (P > 0.05, respectively). However, there was significant correlation between Reffin, Reflex, Refft, Vrin, Vrex and MCA, V5 (P < 0.05, respectively). CONCLUSIONS: The results of four-phase rhinomanometry show significant correlation to acoustic rhinometry. PMID- 19558886 TI - [Therapeutic efficacy of combined treatment with surgical decompression and drug for traumatic optic neuropathy]. AB - OBJECTIVE: To evaluate the therapeutic efficacy of combined treatment with surgical decompression and drug for traumatic optic neuropathy (TON) and analyze the influential factors. METHODS: A retrospective study on 69 patients (70 eyes) with TON treated with optic canal decompression through transnasal endoscopic approach and drug was conducted. The visual acuity was divided into 5 grades: no light perception (NLP), light perception (LP), hand move, count finger, > 0.02, marked as I-V respectively. Of 40 eyes with grade I, 18 eyes received emergency operation for severe optic canal fracture confirmed by CT; 22 eyes received corticosteroid therapy firstly and then operation. Of 30 eyes above grade I, 16 eyes with optic canal fracture confirmed by CT received emergency operation; 14 eyes received corticosteroid therapy firstly and 3 days later received operation. Postoperative follow-up lasted 3-12 months to observe the recovery of visual acuity. RESULTS: The therapeutic efficacy of patients with the visual acuity of LP and above LP was better than that of NLP (90.0% to 27.5%), the difference had statistical significance (chi(2) = 26.98, P < 0.001). In operated group, the therapeutic efficacy in patients whose visual acuity was improved from NLP after glucocorticoid therapy (80.0%) was better than that of the patients with no improvement (5.9%), the difference had statistical significance (chi(2) = 12.09, P < 0.001). CONCLUSION: The imaging findings of optic canal fracture can not be used as determinants for operation. The patients with NLP whose visual acuity had no improvement after corticosteroid therapy are poor candidates for surgical decompression. The visual acuity before treatment is the main factor affecting the therapeutic efficacy. PMID- 19558887 TI - [Fibrotic remodeling of nasal mucosa in rat model after radiation injury]. AB - OBJECTIVE: To investigate the role of nasal mucosa fibrosis on radiation induced nasal mucosa injury. METHODS: Seventy two male rats were randomly divided into two groups, control group and irradiation injured group (radiation dose were 40 Gy); the rats were killed 1 week, 2 weeks, 4 weeks, 8 weeks, 3 months and 6 months after the finish of radiation. The middle turbinates of the animals were removed. The pathological change of the nasal mucosa were observed with scanning electron microscope, transmission electron microscope, hematoxylin and eosin (HE), alcian blue-periodic acid-Schif (AB-PAS), and Masson Trichrome (MT). The Hyp content in nasal mucosa was measured with chemo-chromatometry. RESULTS: After radiation, the pathological characteristics in early stage (within 4 weeks) was acute inflammatory reaction. The repair of nasal mucosa started 4 weeks after radiation, lasted to 6 months. The deposition of collagen in nasal mucosa could be found 1 week after irradiation and increased gradually. CONCLUSION: Irradiation could induce a serials of pathological changes on nasal mucosa. The nasal mucosa fibrosis may be one of the reasons of persistent irradiation induced nasal mucosa injury. PMID- 19558888 TI - [Parathyroid glands are differentiated from lymph node by activated-carbon particles]. AB - OBJECTIVE: To probe into an new methods preserving parathyroid gland of patients with thyroid carcinoma. METHODS: Thirty-six patients with thyroid carcinoma that primary treated were random divided into two groups: trial group and control group. Emulsion of activated-carbon particles was injected into the thyroid gland of trial group patients. After thirty minutes, central compartment dissection was performed in the all patients. The black stained tissue in the dissection specimen of trial group was separated. Total lymph node, metastasis lymph node and parathyroid gland in the black stained tissue, and non-black stained tissue in the central compartment dissection specimen of trial group and central compartment dissection specimen of control group were counted respectively. RESULTS: Nineteen and twenty central compartment dissection was performed in trial group and control group respectively. There are 177 lymph nodes included 83 metastasis lymph nodes in the black-stained tissue of central compartment dissection specimen of trial group. No parathyroid gland was found in the black stained tissue. Nine lymph nodes included 2 metastasis lymph nodes and 7 parathyroid glands were found in the non-black stained tissue of central compartment dissection specimen of trial group. There were 124 lymph nodes included 80 metastasis lymph nodes and 8 parathyroid glands in central compartment dissection specimen of control group. There are statistic difference between the amount of lymph node in black stain tissue and that of control group (t = 0.340, P = 0.003). Rate of staining lymph node were 95.2 percent. CONCLUSIONS: Lymph node of VI group can be stained black by activated carbon particles, and parathyroid gland cannot be stained black. Maybe, parathyroid gland can be preserved by removing the black stain lymph node and retaining the non-black stained tissue. PMID- 19558889 TI - [Diagnostic value of osseous diversity on CT to sinonasal inverted papilloma]. AB - OBJECTIVE: To evaluate the CT characteristics of osseous diversity in patients with sinonasal inverted papilloma (SIP) and to correlate these characteristics with the origins of tumors. METHODS: Sinonasal CT images of 28 patients were reviewed retrospectively to detect areas within which there was focal hyperostosis. The correlation between the sites on the CT scans within which there was osseous diversity and the origin of the tumors in the surgery was analyzed. RESULTS: Surgical evaluation of 22 lesions with focal hyperostosis in CT images revealed that 20 of these lesions coincided with the actual origin of tumor. The focal hyperostosis on CT images corresponded to the actual tumor origin in 71.4% of cases. It denotes the origin of SIP could be predicted from the focal hyperostosis site. The origin of the other 2 cases were disaccord with the focal hyperostosis site, one case had the origin of uncinate process but hyperostosis of anterior middle turbinate, another case had the origin of superior turbinate but hyperostosis of uncinate process. Nine lesions of 28 patients had air sign, 21 lesions had bone absorption and destruction and 19 lesions had bone displacement in CT images. The pressure from tumor growth can induce varied bone destruction and displacement, the source of SIP can be estimated by the bone destruction site and displacement direction. CONCLUSIONS: There has a relatively high concordance between the origin of the SIP and focal hyperostosis on CT, The source of SIP could be discerned indirectly by bone destruction and displacement, these conduce to assess the disease before surgery and make pertinent operation. PMID- 19558890 TI - [Correlation between reduction of distortion product otoacoustic emission and percentage of outer hair cell missing in chinchillas]. AB - OBJECTIVE: To explore the quantitative relationship between the reduction of distortion product otoacoustic emission (DPOAE) and the percentage of outer hair cell loss. METHODS: Coadministration of cisplatin (0.2 mg/kg) and ethacrynic acid (40 mg/kg) were used to establish a cochlear lesion model in chinchillas. DPOAE was measured before and 1 week, 2 weeks, and 3 weeks later respectively after cisplatin and ethacrynic acid treatment. Animals were terminated 3 weeks after the treatment. Cochlear surface preparations were performed, and the cochlear hair cells were counted through entire length of the cochlea. The correlation between DPOAE reduction and outer hair cell missing was analyzed using Pearson correlation analysis. RESULTS: Cisplatin and ethacrynic acid treatment induced cochlear hair cell lesion that the outer hair cell loss in the cochlea developed in a stereotypic pattern; damage began in the base of the cochlea and progressed towards the apex. Reduction of DPOAE was relatively consistent with outer hair cells loss. On the average, 1% outer hair cells loss may result in 0.24 dB reduction in DPOAE levels. Pearson analysis showed a positive correlation between the reduction in DPOAE and missing of outer hair cells (r = 0.796, P < 0.05). CONCLUSIONS: It may be helpful to evaluate missing percentage of outer hair cells from reduction in DPOAE levels. PMID- 19558891 TI - [Edaravone protects hearing from acute acoustic trauma in guinea pigs]. AB - OBJECTIVE: To assess whether the free radical scavenger, edaravone, could provide protection from oxidative stress and hearing loss induced by noise exposure. METHODS: Forty-eight guinea pigs were divided into six groups and exposure to a stationary noise at 125 dB SPL for 2 h only once. Group A: measured hearing and reactive oxygen species (ROS) level without noise exposure. Group B: intratympanic saline injection. Group C: intratympanic edaravone injection. Group D: exposed to noise for 2 h. Group E: intravenous edaravone injection after noise exposure. Group F: intratympanic edaravone injection after noise exposure. All animals of group D, E and F were measured hearing with ABR before noise exposure, immediately after noise exposure and at 2, 6, 12, 24, 48, 72 h after noise exposure, and then were decapitated for ROS measurement with electron spin resonance technology. RESULTS: After noise exposure, the ABR threshold of group D increased immediately after acute acoustic trauma and did not get right finally, while group F came back about 10 dB SPL. The normal level of ROS in the cochlea of guinea pigs was about 21.68 (cm/g) and significantly increased after acute acoustic trauma, reaching its peak in 2h, and didn't return to normal after 72 h. Group E did not inhibit free radicals, while group F showed significant effect on inhibiting production of free radicals. CONCLUSIONS: The level of ROS in cochlea were decreased significantly after intratympanic edaravone injection. The mechanism may due to its effective clearance of the ROS in cochlea. PMID- 19558893 TI - [Real-time Taqman probe technique system for detecting the MtDNA 1555 A > G mutation.]. AB - OBJECTIVE: To establish a Real-time Taqman probe technique system to detect the mtDNA 1555A > G mutation in deaf population. METHODS: Primers and Taqman probes for mtDNA 1555A > G mutation were designed and synthesized. The technique system for detecting mtDNA 1555A > G mutation using Real-time Taqman probes was established. Then the reliability of the technique was tested in 132 patients with severe to profound hearing loss who were detected for the mtDNA 1555A > G mutation by sequencing, Kit method and Real-time Taqman probe technique at the same time. Finally, the results by the above three ways were compared. RESULTS: Thirty-two cases with mtDNA 1555A > G mutation were found by the technique of Real-time Taqman probe. These findings coincided with the results from sequencing and Kit method completely. Both the false positive rate and the false negative rate were zero. CONCLUSIONS: The technique possesses the merits of accuracy, convenience, high sensitivity, high specificity and intuitionistic results, etc. Importantly, the Real-time Taqman probe technique only needs 1.5 hours to detect the 1555A > G mutation and it saves 4.5 hours for one reaction compared with the Kit method popularly used nowadays. The technique system of detecting mtDNA 1555A > G mutation is reliable. It's suitable for large-scale detecting and preventive diagnosis of mtDNA 1555A > G mutation. PMID- 19558892 TI - [Role of caspase 12 in apoptosis of cochlea induced by intense noise in guinea pigs]. AB - OBJECTIVE: To investigate the relationship between caspase 12 activation and endoplasmic reticulum stress mediated apoptosis of guinea pig cochlea cells induced by intense noise. METHODS: Thirty-two guinea pigs were randomly divided into 4 groups. The guinea pigs in the experiment groups were exposed to 4 kHz narrow band noise at 120 dB SPL for 4 h. After the noise expose for 1, 4, 14 days of the experiment guinea pigs, auditory brainstem response (ABR) of the guinea pigs on experiment and control groups were tested before decapitated. Four guinea pig's cochleae of every group were taken to paraffin section, and the rest was extracted the total protein. Apoptosis was tested by terminal deoxynucleotidyl transferase (TDT)-mediated deoxyuridine triphosphate (d-UTP) nick and labeling method (TUNEL) and transmission electron microscopy. Expression of caspase 12, Bip/GRP78 was tested by immunohistochemistry and Western blot methods. RESULTS: The observation by transmission electron microscopy showed the features characteristic of apoptotic cells in the Corti and SGC of 1d after the noise expose, but no in the control. There were higher expressions of Tunel-Positive cells in the OHC, SGC and SV of experiment groups, and there was significant differences compared with the control group (P < 0.01). Protein levels of Bip/GRP78 and caspase 12 were risen up after noise exposed, and there all were significant differences compared with the control group (P < 0.01). CONCLUSION: Intense noise causes cochlea cell lesion by inducing apoptosis to result in and caspase 12 induced endoplasmic reticulum stress-related apoptosis plays an important role in the procedure of apoptosis. PMID- 19558894 TI - [Significance of three imaging examinations performed before decompression surgery for traumatic optic nerve injury]. PMID- 19558895 TI - [Gene mutation and cochlea environment]. PMID- 19558896 TI - [An especially large and long-term tracheal foreign body]. PMID- 19558897 TI - [Diagnosis and treatment of laryngopharyngeal reflux disease]. PMID- 19558898 TI - Protective effect of glycyrrhizin on nephrotic syndrome induced by adriamycin in rats. AB - Purpose: To explore the protective effect of glycyrrhizin in rats with nephrotic syndrome (NS) induced by adriamycin (ADR). Methods: 36 Sprague Dawley (SD) male rats were divided into control, untreated and glycyrrhizin treatment groups. The NS rat model was established by injecting ADR twice in the untreated and in the glycyrrhizin treatment groups. Rats in the glycyrrhizin treatment group were fed glycyrrhizin by intragastric administration for 7 days. Changes in the following indices were observed in the three groups before and 4 weeks after the treatment: 24h urine protein quantitation (UPr), serum cholesterol (Ch), serum albumin (Alb), blood urea nitrogen (BUN), serum creatinine (sCr), laminin (LN), fibronectin (FN), collagen (Col), transforming growth factor ?1 (TGF?1) and connective tissue growth factor (CTGF); histopathology by light and electron microscope. Expression of LN, FN, Col?, TGF?1 and CTGF in the cortex of the kidney were detected by semi-quantitive immunohistochemical analysis. Expression of TGF?1 and CTGF in the cortex of the kidney was detected by Fluorescein Based Quantitive RT-PCR. Macrophage infiltration was evaluated by the immunoperoxidase staining. Results: Compared with the control group, 24h UPr, Ch, BUN and sCr of rats in the untreated group were increased. Glycyrrhizin reduced 24h Upr, Ch, BUN, sCr, LN, FN, Col, TGF?1, CTGF, and mean arterial blood pressure. Pathological changes in the kidney, the expression of LN, FN, Col, TGF?1 and CTGF in the cortex of the kidney in the glycyrrhizin treatment group were decreased compared with the untreated group. Glycyrrhizin also suppressed macrophage infiltration in the kidneys of NS rat models. Conclusion: Glycyrrhizin exerts protective effects in rats with NS, reducing the excretion of Upr, Ch, BUN, sCr, and mean arterial blood pressure, and also decreasing expression of LN, FN, Col, TGF?1 and CTGF in the kidney. Renal function is improved and the severity of NS is lessened. PMID- 19558899 TI - [Value of dynamic postural control tests on elderly people with vestibulopathy]. AB - INTRODUCTION AND OBJECTIVES: The stability limits and rhythmic weight shift tests study the functional capacity to achieve voluntary postural control of movement in the standing position. The objectives of this paper are to know the interest of these tests in the evaluation of elderly people with vestibular disorders and their relation with the number of falls suffered during the year prior to the study. MATERIAL AND METHODS: Sixty elderly people (65-80 years old) with vestibular disorders (patients) and 60 healthy subjects (control group) of similar age were selected. According to videonystagmographic and clinical criteria, the patients group was divided into compensated and decompensated. All the subjects in the sample performed the stability limits and rhythmic weight shift tests with the NedSVE/IBV system. The number of falls of each subject was determined by a meticulous anamnesis. RESULTS: Compensated patients, decompensated patients and the control group had similar scores in this instrumental functional evaluation, without any statistically significant differences. None of the parameters assessed in this study correlated statistically with the subjects' number of falls during the year prior to the study. CONCLUSIONS: The stability limits and rhythmic weight shift tests are of little utility in the functional evaluation of the elderly with vestibular disorders and in the detection of patients with greater risk of falls. PMID- 19558900 TI - [Clinical symptoms of equilibrium disorders in patients with whiplash syndrome]. AB - INTRODUCTION AND OBJECTIVE: There are many entities that cause equilibrium disorders. Whiplash syndrome is becoming an important entity as a trigger of equilibrium disorders because of an increase in traffic accidents. There are many hypotheses on the generation of vertigo and dizziness in whiplash syndrome. The objective of this study is to describe and analyze the clinical symptoms of patients who suffered whiplash. MATERIAL AND METHOD: Thirty six patients with equilibrium disorders who suffered whiplash syndrome were studied prospectively. None of these subjects had cranial trauma or a history of vestibular pathology prior to the traffic accident. We conducted an exhaustive anamnesis, videonystagmography and cervical magnetic resonance. Patients were classified by type of equilibrium symptom and degree of cervical lesion. RESULTS: 55.5 % of patients had a sensation of dizziness associated with postural and cephalic movements, 38.8 % had disequilibrium continuously, and 16.7 % (6 cases) had vertigo. Three of this last group had a diagnosis compatible with benign positional vertigo but this diagnosis was confirmed in only 2 patients; two patients had labyrinth commotion and one patient had vertigo of unknown origin. CONCLUSIONS: In patients with whiplash, the most frequent equilibrium symptom is the sensation of fleeting dizziness associated with head movements, while only a small group suffer from vertigo. Although vestibular tests are normal in most patients, we cannot rule out the existence of otolithic lesions. PMID- 19558901 TI - [Results of a functional magnetic resonance study of the primary auditory cortex (I): general characteristics and individual outcomes]. AB - OBJECTIVE: To demonstrate and investigate the activation patterns of the primary auditory cortex (Heschl's gyrus) using functional magnetic resonance imaging (fMRI). MATERIAL AND METHODS: Pure tone stimuli at 750 Hz and 2000 Hz were delivered to the right and left ear of 32 normal-hearing volunteers (18-49 years old) in 20-second on-off cycles. The fMRI data were obtained using a 1.5 Tesla scanner and processed with SPM2. RESULTS: For both tone frequencies, bilateral hemispheric activation was identified in the transverse temporal gyrus (Heschl's gyrus) in 29 subjects (90.62 %) in response to pure tone stimuli with a probability level of p < 0.001. For monaural stimulation, bilateral hemispheric activation was observed with generally greater extent of activation in the Heschl's gyrus (HG) contralateral to the stimulated ear. CONCLUSIONS: These results demonstrate that fMRI is a useful imaging technique to investigate the auditory cortex. The contralateral auditory cortex is more responsive than the ipsilateral cortex to tones presented monaurally. PMID- 19558902 TI - [Myringoplasty: auditory follow-up and study of prognostic factors]. AB - OBJECTIVE: To perform a five-year study of hearing evolution in patients undergoing myringoplasty to determine whether eardrum repair correlates with improved hearing. We also studied factors that might predispose to failure and their usefulness as prognostic factors. MATERIAL AND METHOD: In a serial case study, we reviewed all myringoplasties performed at our centre during 2000, 2001, and 2002. We reviewed 83 case histories and studied the closure of the perforation, auditory function pre-operatively, postoperatively and after 5 years, as well as different associated factors. RESULTS: The perforation was closed in 75.9 % of cases. The mean gain of auditory function was 1.5 dB at five years, without statistical significance. The hearing evolution presented a post surgical improvement and a subsequent deterioration, both statistically significant. We found a statistically significant relationship between the size of the perforation and the condition of the contralateral ear. CONCLUSIONS: Perforation closure in our series (75.9 %) is similar to that reported in the literature. We found contralateral ear pathology and the perforation extension to be associated with poor prognosis after myringoplasty. PMID- 19558903 TI - [Results of an organ conservation protocol in patients with locally advanced laryngeal tumours]. AB - INTRODUCTION AND OBJECTIVES: Advanced laryngeal carcinoma patients, candidates for total laryngectomy, nowadays have different treatment options. One of them is induction chemotherapy, which allows modulation of the second manoeuvre according to the degree of response achieved. This study presents the results of a large series of patients treated according to this protocol. METHODS: Retrospective study of 370 patients with T3-T4 carcinomas of the larynx considered as candidates for total laryngectomy. All patients were treated with the protocol mentioned above. RESULTS: Seven patients died as a consequence of the treatment with chemotherapy. Among 363 patients finishing chemotherapy, 154 (43 %) achieved complete response and proceeded with radiotherapy. Total laryngectomy was performed in 135 (37 %) non-responders and 74 (20 %) patients were treated with radiotherapy in spite of an incomplete response to chemotherapy. Overall actuarial survival was 73 % at five years. Survival for patients with complete response and treated with radiotherapy was 77 % and for non-responders subjected to total laryngectomy it was 76 %. Non-responders treated with radiotherapy had a 5-year survival rate of 64 %, significantly different to both other groups (p = 0.01). Larynx preservation was achieved in 42 % of the patients, rising to 72 % for complete responders to chemotherapy and to 62 % in non-responders receiving radiotherapy. CONCLUSIONS: Our larynx preservation protocol including induction chemotherapy for patients with advanced carcinomas who are candidates for total laryngectomy achieved a total survival rate of 73 % with a 42 % larynx preservation rate. The response to the induction chemotherapy was the factor contributing most to larynx preservation. PMID- 19558904 TI - [Fistula following total laryngectomy. Retrospective study and bibliographical review]. AB - BACKGROUND: The pharyngocutaneous fistulae is troublesome and the most common complication following total laryngectomy. Our objective was to determine the incidence of pharingocutaneous fistulae after the total laryngectomy in our serie and to make review of the medical literature. METHODS: We made a retrospective study of a serie of 81 consecutive cases of laryngeal carcinoma treated between 1995 and 2008 in our section. Total laryngectomy was performed in 29 cases and 52 patients treated with organ preservation approach, were excluded. In 14 cases, the procedure was combined with radical neck dissection, pharyngeal resection or myocutanenous flaps. Nasogastric tube for feeding in the postoperative period was used in all patients and surgical gastrostomy was performed in 5 cases. RESULTS: Our incidence of fistulas when total laryngectomy was the alone procedure is 20 % and 34.5 % when simultaneous surgical proceedings were associated. Spontaneous closure was noted in 80 % of the cases and the mean hospitalization time was 23 days. CONCLUSIONS: Most of the fistulas can be managed with conservative treatment. Pectoralis major myocutanenous flap is appropriate when conservative treatment has failed. In small fistulas, nasogastric or gastrostomy tube for feeding can be successfully managed in the ambulatory follow up. The cost-benefit relation must be better analyzed. PMID- 19558905 TI - [Foreign bodies in maxillary sinus]. AB - INTRODUCTION: Foreign bodies in maxillary sinus (FBMS), whatever their origin or nature, are an unusual clinical condition. Diagnosis is based on the radiological findings in a clinical context of unilateral chronic rhinosinusitis. Treatment is the surgical removal of the intrasinusal foreign body. PATIENTS AND METHODS: To identify FBMS, the records of 68 patients with unilateral chronic rhinosinusitis operated on from 2000 to 2007 were reviewed. RESULTS: From 68 records reviewed, we found 11 (16 %) FBMS. Ten (91 %) of these 11 foreign bodies were thought to come from the teeth and the last 1 (9 %) had a non odontogenic origin. Eight of the 11 (73 %) patients with FBMS presented with chronic maxillary sinusitis symptoms and all patients showed radiological findings. Treatment was the surgical removal of the foreign body, in 9 (82 %) patients through endonasal approach by functional endoscopic sinus surgery (FEES) and in the other 2 (18 %) patients a mixed surgical procedure by endonasal meatotomy and oral antrotomy was required. CONCLUSIONS: Chronic maxillary sinusitis showing FBMS is rare and it must be suspected with a prior history of dental procedures. The most frequent source of FBMS is material of odontogenic origin, and non-odontogenic origin secondary to an external injury in an accident or assault is much more unusual. We also review the nature of these foreign bodies, their clinical implications and treatment options. PMID- 19558906 TI - [Clinical significance of unilateral tonsillar enlargement]. AB - INTRODUCTION: Asymptomatic unilateral tonsillar enlargement is usually treated with systematic tonsillectomy under suspicion of malignancy. Due to the fact that most of the cases are benign pathologies, we set out to study the clinical signs that would help us in the diagnosis in order to avoid unnecessary tonsillectomies. MATERIAL AND METHODS: We reviewed 267 tonsillectomies performed from 1996 to 2006 and 30 of these were indicated because of asymmetry. We evaluated risk factors for malignancy: cervical lymphatic node enlargement, sex, age, tonsillar enlargement noticed by the patient, suspicious appearance, systemic symptoms, history of malignancy and immunocompromise. RESULTS: Histopathologic study revealed 80 % to be benign and 20 % malignant. The risk factors with the strongest association were enlargement of cervical lymphatic nodes and suspicious appearance of the tonsil. CONCLUSIONS: Strict control of a unilateral tonsillar enlargement is possible, but it is mandatory to perform a tonsillectomy when the appearance of the tonsil raises suspicions or there are enlarged lymphatic nodes. PMID- 19558907 TI - [Minor salivary gland tumours: a 10-year study]. AB - Salivary gland tumours represent between 2 % and 6.5 %, approximately, of all head and neck tumours. The aim of this paper was to identify the frequency of minor salivary gland tumours among patients in the Oral Medicine Clinic of the Federal University of Parana during the period from 1997 to 2007. A retrospective study was conducted on 1,923 histopathological analyses of oral lesions. Fourteen cases of salivary gland tumours were found, of which 7 were benign and 7 malignant. The lesions were localized mainly in the palate (71.5 %). By histological type, 50 % of the lesions were characterized as pleomorphic adenoma, 28.6 % mucoepidermoid carcinoma, 14.3 % cystic adenoid carcinoma and 7.1 % as polymorphous adenocarcinoma. These findings suggest that salivary gland tumours have a low incidence in the population and that the pleomorphic adenoma is the most common type of tumour, followed by mucoepidermoid carcinoma. PMID- 19558908 TI - [Obstructive sleep apnoea syndrome in children]. AB - Obstructive sleep apnoea syndrome is a well-known clinical entity in adults but until now it has been less well studied in children. In recent years there has been a dramatic increase in the recognition of sleep disorders in children. Our goal is to analyze scientific data published in the last few years. We reviewed published articles regarding paediatric obstructive sleep apnoea syndrome and extracted the clinical symptoms, diagnosis and treatment options. In conclusion, the natural course and long-term prognosis of childhood obstructive sleep apnoea syndrome are not well-known and further studies are needed in this area. PMID- 19558909 TI - [PFAPA syndrome]. AB - PFAPA syndrome is characterized by periodic fever, pharyngitis, cervical adenitis and aphthous stomatitis. The bouts of fever can last for days or even weeks. Between crises, patients remain asymptomatic for variable periods. It appears before the age of five and has limited duration (4-8 years). Its aetiopathogeny is unknown. Corticoids are the treatment of choice. Tonsillectomy has been proposed as a solution but remains controversial. We present the case of a 4-year old girl with PFAPA syndrome who underwent tonsillectomy in January, 2008, and we review the literature. PMID- 19558910 TI - [Simultaneous bilateral facial palsy]. AB - Simultaneous bilateral facial palsy (SBFP) is an uncommon disorder that usually results from a systemic disease, with only a few cases diagnosed as Bell's Palsy. The most common causes of SBFP are head injuries, Bell's Palsy, Lyme disease, Guillain-Barre syndrome, sarcoidosis and meningitis. We present a case of SBFP. PMID- 19558911 TI - [Otic myiasis. Case report]. AB - Otic myiasis is rare in developed countries and is related to environmental conditions and poor hygiene. We present the case of a 65-year-old woman with pain and tinnitus in the left ear lasting for one week. Physical examination in the emergency room showed numerous dipteran larvae totally occupying the external auditory canal. After the diagnosis of otic myiasis, the larvae were removed and the patient was discharged after 24 hours of medical observation. The diagnosis of otic myiasis is by means of direct visualization of the larvae in the ear cavities and treatment includes the removal of larvae from the ear. Prognosis is generally good. PMID- 19558914 TI - [Probiotics in dermatology]. PMID- 19558912 TI - [Calcification of the epiglottis]. PMID- 19558915 TI - [Photodepilation]. AB - Unwanted hair is a common problem for which different types of light therapy have been developed as the treatment of choice. Since 1996, when the American Food and Drug Administration approved the first laser therapy for depilation, much progress has been made in light-based technology and lasers. Lasers and intense pulsed light sources with red or near infrared wavelengths (600 to 1200 nm) are the most widely used for removing hair as they target the melanin of the hair shaft, hair follicle epithelium, and hair matrix. The aim of this review is to describe and compare the different photodepilation methods currently available. PMID- 19558916 TI - [Centenary of the Fontilles Sanitorium]. AB - In recognition of the centenary of the Fontilles Sanitorium, we present some details of its history. The article discusses the foundation of the sanitorium by some of the numerous lepers with no health coverage in the region of Valencia, and Alicante in particular. After a difficult period between 1909 and 1932, the sanitorium was seized by the Republican government. After the civil war, it was returned to the board of trustees, who entrusted administrative tasks to the Compania de Jesus while health care was overseen by the National Health Board. This coexistence was uneasy, and the board of trustees took over again in 1943. We comment on the transition from a sanitorium and colony into a hospital providing health care, research and training, and treatment with sulfones and subsequently other effective drugs. Also discussed are its role in the elimination of leprosy from Spain, admission to the International Federation of Antileprosy Associations in 1969, and its projects in endemic countries, with the ultimate goal of achieving a world free of leprosy. PMID- 19558917 TI - [Electronic medical history: experience in a dermatology department]. AB - Computerization, with a change from paper to electronic format, represents an alternative to traditional information management. This model offers advantages in legibility, uniformity, accessibility, and use of the data. However, it is not easy to apply this process to clinical practice as it requires a suitable network, continuous application development, an implementation strategy, and the cooperation of all staff involved. We have reviewed our experience in the development and introduction of electronic health records and their adaptation to a pioneer dermatology department in Spain. Since our hospital was opened 1998, the model used is that of a single, centralized electronic health record, with supplementary departmental attributes. The electronic health record is conceived as an interactive database designed around the patient, with a procedure-based structure, and that obviates the need for hardcopies (paper or films) in practically all situations; it must comply with legal requirements. The system is installed on central servers maintained by the information technology department. The potential is unlimited; particularly important possibilities include clinical guideline-directed care, remote connection for general practitioners, and online activity, stock, and quality management. With the aim of realizing this potential, a technological change was started in 2003, moving towards what was to become the chosen system in the Community of Madrid to cope with the workload arising from new hospitals. PMID- 19558919 TI - [Multiple primary melanoma]. AB - INTRODUCTION: The risk of multiple melanoma is estimated to be between 1 % and 8 %, with the majority of studies being carried out on North American populations. Our objective was to determine the risk and the clinical-pathological features of multiple primary melanoma in a Spanish Mediterranean population. MATERIAL AND METHODS: We performed a retrospective study of the medical records and the database of the Melanoma Unit of Hospital de Bellvitge, Barcelona, Spain, between 1988 and 2005. RESULTS: We found 25 cases of multiple primary melanoma among 934 patients studied, representing a risk of 2.6 % in our population of melanoma patients. In 50 % of cases, the second melanoma appeared during the first year of follow-up. These subsequent lesions occurred at a different site from the initial lesion in 58 % of cases. In the majority of cases, lesions in a single patient showed similar cytological and architectural features. However, we did observe marked interindividual variability in the histology of multiple primary melanomas. CONCLUSION: Although the risk of a second melanoma in our population appears to be lower than in North American populations, it is not negligible. Melanoma patients must therefore be followed up for life, not only for the risk of metastases but also for the risk of a new primary tumor. Complete examination of the skin must be performed at each visit. PMID- 19558918 TI - [Spanish evidence-based guidelines on the treatment of moderate-to-severe psoriasis with biologic agents]. AB - Psoriasis vulgaris is an inflammatory skin disease that is generally chronic and that affects between 1 % and 2 % of the population in industrialized Western countries. It is associated with a marked decline in quality of life. A wide range of treatments are currently available, although surveys conducted before the advent of biologic agents reflected a strong degree of dissatisfaction with the treatments then available. Extensive scientific evidence has been gathered on the safety of biologic agents, and this has led to a review of the role of systemic treatment in general and has allowed new therapeutic goals and strategies to be contemplated in patients with moderate-to-severe psoriasis. In this new situation, there is a need for Spanish guidelines on the treatment of moderate-to-severe psoriasis with biologic agents, drafted by consensus among specialists and ratified by the Spanish Psoriasis Group of the Spanish Academy of Dermatology and Venereology (AEDV). These guidelines should be evidence-based with regard to the pharmacologic characteristics, mechanism of action, administration route and regimen, efficacy, contraindications, adverse effects, and cost estimates of biologic agents approved for the treatment of moderate-to severe psoriasis in Spain. PMID- 19558920 TI - [Sequential histological and immunohistochemical assessment of proliferation and apoptotic markers during treatment of psoriasis with antitumor necrosis factor alpha (infliximab)]. AB - BACKGROUND AND OBJECTIVES: Psoriasis is an inflammatory skin disease of immunologic nature that is mediated by T-helper-1 cytokines. Clinical response to treatment with antitumor necrosis factor (TNF) alpha antibodies (infliximab) has been significant; however, the mechanisms for clearance of lesions have not been elucidated. The aim of the present study was to assess variations in the histology and expression of proliferation and apoptotic markers in sequential skin biopsies of patients with psoriasis treated with infliximab. MATERIAL AND METHODS: We studied skin biopsies (of lesioned and healthy skin) from 3 patients with extensive moderate-to-severe psoriasis (mean psoriasis area and severity index [PASI] score, 35) treated with intravenous infliximab infusions (5 mg/kg) at weeks 0, 2, and 6. Biopsies were taken on days 0, 14, and 28, and were processed for conventional histological and immunohistochemical study. The apoptotic markers used were TP53, B-cell lymphoma 2 protein, anticaspase 3, and anticaspase 8. The cell proliferation marker used was Ki67. RESULTS: Treatment with infliximab was associated with a significant clinical improvement in 3 patients (mean PASI score, 21.6 at 14 days and 13.9 at 6 weeks), which correlated with the progressive disappearance of histological lesions with a decrease in epidermal proliferation. However, apoptosis was not observed, and the samples tested negative for anticaspase antibodies. Expression of TP53 decreased 2 weeks after starting treatment, and was similar to that in normal skin at 28 days. CONCLUSIONS: Clinical and histological response of psoriasis to infliximab was not associated with a significant increase in the apoptotic markers assessed. PMID- 19558921 TI - [Bilateral symmetrical nail hypoplasia of the hands]. PMID- 19558922 TI - [Necrotic papules associated with hemoptysis and acute renal failure]. PMID- 19558923 TI - [Perforating lichen nitidus on an atypical site]. PMID- 19558924 TI - [Eczema herpeticum in cutaneous T-cell lymphomas]. PMID- 19558925 TI - [Verrucous carcinoma of the foot associated with human papilloma virus type 18]. PMID- 19558926 TI - [Cold urticaria associated with Epstein barr virus mononucleosis]. PMID- 19558927 TI - [Verrucous hemangioma versus microcystic lymphatic malformation]. PMID- 19558929 TI - [Disseminated pyogenic granuloma induced by erythropoietin?]. PMID- 19558930 TI - [Who teaches us how to investigate?]. PMID- 19558931 TI - [Gender differences in the hospitalization and reperfusion delays in the acute coronary syndrome]. AB - OBJECTIVE: To study delay times in women with acute coronary syndrome (ACS) and ST segment elevation (STSEACS) until administration of possible reperfusion therapy as well as its possible differences in relationship to the men. MATERIAL AND METHODS: A study was conducted in 1,849 patients consecutively hospitalized in a Coronary Unit from January 2000 to December 2005 who had been diagnosed of ACS. Different delay times were studied from the beginning of the symptoms, comparing them between genders and correcting this by variable age. RESULTS: The percentage of women was 24.4% with a mean age of 71.5 years. Medium delay time from the beginning of the symptoms to contact with the first health care agent was significantly (p < 0.001), longer for women than for men, 164 min. vs 120 min. Reperfusion treatments were used in 57.6% of the subjects, with a significant difference (p < 0.01) of 7.6% between men (59.5%) and women (51.9%). CONCLUSIONS: Delay times are greater in women than men, above all regarding the beginning of the symptoms until arrival to the first health care agent. Currently, reperfusion treatments are performed more in men than in women, the differences are minimum when adjusted by age. PMID- 19558932 TI - [Needs of the families in a Critical Care Unit]. AB - OBJECTIVES: To identify the needs of the family of admitted patients in the Critical Care Unit (CCU), and their level of satisfaction with the attention that they received. DESIGN: Cross-sectional, observational study. SETTING: Critical Care Unit, Virgen Macarena University Hospital, Seville. SUBJECTS OF THE STUDY: Two-hundred and sixty-eight families were selected through a random sampling system. INSTRUMENTS: A survey was developed using bibliography as reference. RESULTS: According to information, it was observed that receiving information after visiting hours and its access was difficult in regards to unforeseen changes in the patient's course. On the other hand, there were high levels of satisfactions with the attention received and with the nursing cares. Nevertheless, several deficiencies were observed in the identification of all the professionals, the visiting hours, the physical conditions of the waiting room, and privacy within patient's care environment. CONCLUSIONS: One of the principal needs of the relatives was to spend more time with the patients. Regarding to the information, access after visiting hours, and on that regarding unforeseen changes in the patient's evolution should be reviewed. Regarding satisfaction, in general, families were highly satisfied with the attention received, especially with the nurse's care. Nevertheless, these professionals should encourage the participation of the families in the patient's care. Moreover, all the areas of improvement observed should be dealt with urgently, mainly the visiting hours. PMID- 19558933 TI - [Evaluation of the effectiveness of an in-hospital cardiac alarm system]. AB - OBJECTIVES: MAIN OBJECTIVE: Evaluate effectiveness of our cardiac arrest (CA) system. SPECIFIC OBJECTIVES: To know the clinical and demographical characteristics of patients who undergo cardiopulmonary resuscitation (CPR). To know the action time of the CA team, immediate survival, hospital discharge and at one year, cerebral functional category of cardiorespiratory arrest survivors. MATERIAL AND METHODS: Retrospective descriptive study on CA received in the Hospital San Millan of Logorono. SUBJECTS: In-hospital cardiac arrest adults except for those in the Intensive Care Unit (ICU) and Emergency Department. PERIOD: From January 1, 2003 to December 31, 2006. Survival study until December 31, 2007. Data was collected from the Utstein Style Registry of the in-hospital CPR. Descriptive statistic analysis and bivariant analysis was performed with the Chi2 test and odds ratio (OR). RESULT: CPR was performed in 90 patients. Average arrival time was 1 minute. Origin was non-cardiac 56%, cardiac 34.4%. Initial rhythm was non-shockable 76.7%, shockable 22.2%. Survival at hospital discharge: 20%. Cerebral Performance Category 1: 89%. Survival after one year 16.7%. Survival: cardiac etiology (P=0.05; OR 0.20; 95% CI 0.06-0.65). Initial rhythm shockable (P=0.0001; OR 0.18; 95% CI 0.05-0.56) CONCLUSIONS: Our CA system is effective. We achieve a survival and cerebral performance categories on hospital discharge that is similar to other studies although the rate of patients with shockable rhythm and cardiac etiology in our series is less than others published. PMID- 19558934 TI - [Percutaneous tracheostomy procedure: nursing service control and follow-up]. AB - A significant number of patients in the Intensive Care Units require ventilator support over long periods of time. A tracheostomy is necessary in these cases so that the patients can breathe properly and to allow access to the airway. Percutaneous tracheostomy (PT) is an increasingly used and minimally invasive procedure in which the Seldinger method is used for access the trachea. In recent years, it has become the alternative to surgical tracheostomy. In fact, it is becoming the technique of first choice in critical patients since it has many advantages regarding the conventional tracheostomy. It is an easy-to-use and safe technique, but one that is not exempt from risks. The nursing staff that helps to perform this operation must know its procedures, advantages, disadvantages, indications, contraindications and complications in order to act correctly and in coordination with the rest of the team and to be able to satisfy the patient's needs at each moment. In this article, the characteristic interventions of the nursing staff before, during and after performing this technique are explained. PMID- 19558935 TI - [Use of non-invasive ventilation in acute respiratory failure. Multicenter study in intensive care units]. AB - OBJECTIVES: Study the use of non-invasive ventilation (NIV) in patients with acute respiratory failure in intensive care units (ICUs) in Spain. METHODS: A questionnaire was sent to 254 ICUs, after which, they were invited to participate in a multicenter, retrospective study, providing detailed information on ventilated patients. RESULTS: Answers were received from 123 hospitals. Of these, 119 used NIV, although its use varied greatly. NIV is the treatment of choice in 89% of the units for chronic obstructive pulmonary disease (COPD), in 79% for acute pulmonary edema (APE), in 53% for postextubation failure, in 53% for pneumonia 53%, and in 17% for acute respiratory distress syndrome (ARDS). It was used occasionally in COPD in 11% of the units, and in 21% of the units for APE. Eighteen hospitals provided additional information on 432 ventilated patients, 232 (54%) of whom received NIV as first line therapy. Presence of pneumonia or acute respiratory distress syndrome (ARDS) was an independent predictive factor of NIV failure (ORa=5.71; CI 95%, 1.83-17.8; p=0.003). Admission in a unit with experience in NIV in >50 patients/year (ORa=0.22; CI 95%, 0.07-0.63; p=0.005) and a higher PaO2/FiO2 ratio after one hour of ventilation (ORa=0.98 per point; CI 95%, 0.97-0.99; p<0.001) were protector factors. CONCLUSIONS: In Spain, NIV is widely used but it may continue to be underused in COPD and APE. The diagnosis of pneumonia or ARDS was an independent predictive risk factor. Admission in an ICU with NIV in more than 50 patients/year also have higher PaO2/FiO2 ratio after one hour of ventilation were predictive factors of success. PMID- 19558936 TI - [Outcome of the critical patient according to the sex and the age]. AB - OBJECTIVE: To determine if gender and age are associated with differences in mortality in patients requiring critical care. DESIGN: Retrospective analysis of prospectively collected data over 6 consecutive years. SETTING: Polyvalent intensive care unit (ICU) of a tertiary hospital in the Canary Islands. PATIENTS: Adult patients who were hospitalized in the ICU, divided on the basis of gender and age (or=65 years). PRIMARY VARIABLES OF INTEREST: Demographic and clinical diagnosis data on ICU admission, APACHE II, days of mechanical ventilation, days of renal replacement therapy (RRT) and the mortality were collected. RESULTS: During the study period, 3786 patients were admitted to the ICU, 66.7% male and 2469 (65.2%)<65 years. Mortality in the ICU of the coronary group was greater in women (11.1% vs 6.7%; p=0.02), even though there were no significant differences between both genders in the APACHE II (p=0.56). No statistically significant differences were found according to gender in age (or=65 years), in the APACHE II, or in the need for mechanical ventilation or renal replacement therapy as well as in the ICU mortality. We also found no differences in the mortality between the diagnostic groups and gender even when the APACHE II was significantly greater on admission, as occurred for the women<65 years of the coronary and traumas group and for women with surgery>or=65 years. CONCLUSIONS: No significant differences were demonstrated in the outcome in relationship with gender except for in the coronary group in which mortality was greater in women. Age above or below 65 years had no influence on mortality in our patients. PMID- 19558939 TI - [Full stop and continuing]. PMID- 19558937 TI - [Prognostic value of lactate clearance in the first 6 hours of intensive medicine course]. AB - OBJECTIVE: Analyze the clinical usefulness of lactate clearance (CL6) immediately after admission to the intensive care unit (ICU) in the first 6 hours. SETTING: Surgical-ICU. Centro de Asistencia del Sindicato Medico of Uruguay. DESIGN: Prospective, analytic and observational study performed between December 1, 2004 and March 31, 2006 in patients over 18 years whose arterial lactate level is higher than 2 mEq/l on admission to the ICU. Lactate clearance (CL6) was defined as the quotient between admissions (L0) minus the six hour lactate level (L6) divided by the admission lactate level. Sensitivity, specificity, positive and negative prognostic value for different CL6 cutoff were analyzed. The optimal CL6 was considered as the cutoff with the highest sum of sensitivity plus specificity. RESULTS: One hundred and eight patients were included; 64 patients died (mortality intra-ICU 59.3%). ICU mortality related variables, identified by Cox regression analysis, were CL6 (HR=0.458; CI 95%, 0.239-0.876), L0 (HR=1.16; CI 95%, 1.033-1.303) and SAPSII (HR=1.019; CI 95%, 1.006-1.034). A CL6 equal to or lower than 0.4 was considered as optimal cutoff with a positive prognostic value of 74% and negative prognostic value of 61%. It was also associated with lower survival adjusted by the SAPSII value and L0. CONCLUSIONS: In critically ill surgical patients, whose CL6 on admission was over 2 mEq/l, lactate clearance in the first six hours could be useful to predict the ICU outcome. PMID- 19558938 TI - [Prognosis of the sequels after brain injury]. AB - Acquired brain injury (ABI) is a major public health problem due to its high incidence and prevalence, long-term effects on patients and their families and enormous socioeconomical costs. In our country, this is treated unequally by the different institutions and specialties. Its etiology, by order of incidence, is due to stroke, traumatic brain injury and anoxic-ischemic encephalopathy and then, at a great distance, a miscellaneous group in which hypoxic encephalopathy stands out. ABI has two extreme poles: deep coma and full reintegration into the community with a similar level as prior to the lesion. Between these poles are the vegetative states, minimally conscious states and, when there is minimal cognitive recovery, a varying range of difficult-to classify impairments, disabilities and handicaps, due to their extreme heterogeneity. The long-term outcome is assessed by descriptive and functional scales, which usually have important feasibility and validity problems. Some scales (GOS, GOSE) classify functional deterioration during the acute and subacute lesional phase. Others analyze neurorehabilitation planning and monitoring (ERLA, Barthel Index). The International Classification of Impairments, Disabilities and Handicaps (ICIDH-2) describes disabilities and impairments. The efficacy of rehabilitation treatment depends on the how early they are done, their adaptation to each patient's needs, intensity and performance by qualified rehabilitation centers. It is difficult to quantify their results in order to compare them because of the serious methodological difficulties. PMID- 19558940 TI - [Brain death diagnosis]. AB - Brain death has been recognized by the scientific community as the person's death, and accepted in the legislation of different countries. Brain death is defined as the irreversible ending of the functions of all the intracranial neurological structure in both the brain and brain stem. This clinical situation appears when intracranial pressure exceeds the patient's systolic blood pressure, leading to brain circulatory arrest. The most frequent are cerebral hemorrhage and cranioencephalic trauma. Clinical diagnostic must be done by doctors with expertise in neurocritical patient treatment. This diagnosis is based on a systematic, complete and extremely rigorous clinical examination that confirms a non-reactive coma, absence of brain stem reflex, and absence of spontaneous breathing. Instrumental tests may be obligatory in some cases, this depending on each country. Electroencephalogram and evoked potentials are the electrophysiological tests used. In patients treated with sedative drugs, cerebral blood flow evaluation tests, such as cerebral angiography, transcranial Doppler or 99Tc-HMPAO scintigraphy, will be used. More than 92% of the transplants performed in Spain are performed with brain death donor organs. Brain death confirmation is a high responsibility act, with medical, ethical and legal significance since it requires removal of all artificial support, or organs extraction for transplant. Extensive knowledge on its diagnostic and correct decision making avoid unnecessary use of resources and improves management of organs for transplant. PMID- 19558941 TI - [Bacterial infections in critically ill patients: review of studies published between 2006 and 2008]. AB - A systematic revision of medical publications between 2006 and 2008 regarding bacterial infections that affect the critical patients was performed. Four subjects were selected: Community-acquired pneumonia, ventilator-associated pneumonia, catheter-related bloodstream infection and new antimicrobial treatments. When dealing with community-acquired pneumonia and due to the absence of completely reliable standards, it is necessary to follow the locally adapted guidelines of clinical practice, to identify patients related to the health-care system and admit patients to the ICU in accordance with the criteria. Regarding the etiological diagnosis of ventilator-associated pneumonia, any microbiological information available must be used. Due to the risk of multidrug bacteria, combined empiric therapy should be initiated immediately and then mono-therapy adjusted to the antibiogram should be established. Already established measures for mechanical ventilation associated pneumonia and catheter-related bacteriemias, which have been effective, should be implemented. The empirical treatment of catheter-related bacteremia must be directed towards the most probable pathogens according to the puncture site. The most recently sold antibiotics are basically directed towards multidrug gram positive resistant bacteria. However, for the treatment of gram negative resistant bacilli, the use of the new antimicrobials must be combined with a new evaluation of the antibiotics that have been used for years and the possibility of choosing different administration forms. PMID- 19558942 TI - [Preliminary experience in an intensive care unit with Impella Recover. A new circulatory support system in patients with low cardiac output]. AB - The low cardiac output syndrome following cardiopulmonary bypass is characterized by poor left ventricular contractibility that requires the support of high doses of vasoactive drugs, intra-aortic balloon pump, and sometimes makes it impossible to disconnect the extracorporeal circulation. We report 5 cases in which a "recently created" device in left ventricular support was inserted, the Impella Recover (Impella CardioSystems AG, Aachen, Germany) due to cardiogenic shock at the end of the surgery. Four of these patients recovered their heart function and the ventricular support could be removed after 70+/-55 h. In a fifth patient, the right ventricular failure warranted the use of Berlin Heart assist device. PMID- 19558943 TI - [The risk of delaying admission to the ICU in critical patients]. PMID- 19558944 TI - [Recombinant activated factor VII for spontaneous cerebral hemorrhage]. PMID- 19558946 TI - [Aneurysms and penetrating ulcers of the thoracic aorta]. PMID- 19558947 TI - [The last two years in the SEMN. Balance and future]. PMID- 19558948 TI - [PET/CT with (11)C-choline and (18)F-FDG in patients with elevated PSA after radical treatment of a prostate cancer]. AB - OBJECTIVE: To compare the diagnostic accuracy of PET/CT with (18)F-FDG and (11)C choline for early detection and localization of recurrent prostate cancer. MATERIAL AND METHODS: Thirty-eight patients with increased PSA levels (0.8-9.5 ng/ml) after radical treatment for prostate cancer (surgery n = 20/radiation therapy n = 18) were included. Ten patients were on hormone therapy. All patients underwent a PET/CT with (11)C-choline and (18)F-FDG, respectively, on the same day. The PET imaging findings were compared with histopathology (n = 10); PSA monitoring (n = 21) and/ or other methods (n = 7). RESULTS: Focal uptake of (11)C choline was detected in 26 patients (68%), and focal uptake of (18)F-FDG was detected in 13 patients (34%). The (11)C-choline uptake in 14 patients was suggested local recurrence, whereas this was true in only 4 patients (48%) with (18)F-FDG. Pelvic lymph nodes were detected with (11)C-choline PET/CT in 8 patients and only in 4 patients (50%) with (18)F-FDG. Mediastinal involvement was detected in 5 patients with (11)C-choline and 3 patients (60%) with (18)F-FDG. Focal bone involvement was detected in 3 patients with (11)C-choline and (18)F FDG. (11)C-choline was able to detect 40% of recurrences in patients with PSA < 1 ng/ml, 50% of recurrences in patients with PSA 1-4 ng/ml and 87% of recurrences with PSA > 4 ng/ml. Sensitivity of (11)C-choline was higher for surgically treated patients, with no significant differences found between patients with and without hormone therapy. CONCLUSIONS: (11)C-choline PET/CT was useful for the detection of biochemical recurrence of prostate cancer, with higher yielding as compared to (18)F-FDG. (11)C-choline sensitivity was clearly related to PSA levels, was higher in patients with surgery and did not seem to be modified by hormonal therapy. Disease staging with (11)C-choline showed direct impact for the selection of the most appropriate therapeutic approach. PMID- 19558949 TI - [Impact of (18)F-FDG PET/CT on the therapeutic management in the initial staging of the esophageal cancer]. AB - OBJECTIVE: The incidence of esophageal cancer has increased considerably over recent years, it now being the 6th most frequent cause of cancer-related death. Our study has aimed to compare the clinical value of PET/CT and CT scan in the initial staging of patients with esophageal cancer. MATERIAL AND METHODS: Fifty nine patients (6 women) diagnosed of esophageal cancer were assessed retrospectively. All patients underwent diagnostic CT scan and PET/CT for initial staging within 3 to 15 days following clinical diagnosis. RESULTS: PET/CT showed intracellular (18)F-FDG entrapment having pathological significance in all the tumors (100%), signs of locoregional lymph node infiltration (N1) in 34 and a total of 19 lesions consistent with metastasis (M1) in 14 patients (23.72%). The CT scan detected malignancy in 57 patients (96.6%), abnormal lymph node in 32 patients and 17 N1 in 12 patients (20.33%). In three cases, CT- PET detected synchronous esophageal lesion in staging studies for other neoplastic processes (lung and ear-nose-throat). CONCLUSION: PET/CT showed a higher detection rate of primary malignant lesions, abnormal lymph nodes and distant metastases. A change in stage was only observed in two patients. PMID- 19558950 TI - [Clinical utility of (18)F-DOPA-PET in movement disorders. A systematic review]. AB - INTRODUCTION: (18)F-fluorodopa ((18)F-DOPA) is a functional tracer of presynaptic dopaminergic neuron terminations in the nigrostriatal system. This review is aimed to assess the efficacy of (18)F-DOPA-PET in the diagnosis and evaluation of the progression of Parkinson's Disease (PD) and in the differential diagnosis with other Parkinsonism Syndromes. METHODS: A review was made of the literature by searching six databases and selecting the most relevant articles according to strict inclusion and exclusion criteria. The study data were systematically extracted and included in evidence tables. RESULTS: Of the 1478 registries recovered through the search of the literature, 48 studies were extracted. Of these, only 13 were included in the systematic review. It was observed in all of them that PD is manifested by a lower striatal uptake of (18)F-DOPA, especially in the putamen with posterior predominance. Prospective studies have shown that there is loss of uptake with disease progression. One article described regional differences in (18)F-DOPA striatal pattern between PD, multisystem atrophy (MSA) and progressive supranuclear palsy (PSP). Cognitive impairment in PD seems to be related with (18)F-DOPA abnormal uptake in some regions of frontal cortex and caudate nucleus. CONCLUSION: (18)F-DOPA-PET seems to be useful for the diagnosis and evaluation of PD progression. However, the evidence is not conclusive regarding its utility in the differential diagnosis with other Parkinsonism Syndromes and in the differentiation between ex novo and advanced PD. PMID- 19558951 TI - [Regional cerebral blood flow changes in Parkinson's disease: correlation with disease duration]. AB - INTRODUCTION: Changes in regional cerebral blood flow (rCBF) have been reported in idiopathic Parkinson's disease (PD). Nonetheless, their typical pattern still remains controversial regarding some features, such as basal ganglia involvement and the main cortical regions affected. Functional neuroimaging makes it possible to identify the brain dysfunctions of the neural circuits underlying the disease. Voxel-based analysis methods make it possible to increase the reliability of the results. OBJECTIVE: To assess the rCBF changes in patients with PD and their relation with disease duration. MATERIALS AND METHODS: Thirty PD adult patients without dementia underwent evaluation with (99m)Tc-ECD SPECT. SPM5 was used for statistical comparison with 25 normal controls of similar ages. The disease course duration in years was added as a covariate. Additionally, patients with a 6-year evolution or less and those with more than 6 years were compared separately with normal controls. RESULTS: Significant hypoperfusion was detected in bilateral premotor and posterior parietal cortex and increase of perfusion was present in the cerebellum. These changes correlated with the years of evolution of the illness. Patients with longer evolution also presented thalamic, subthalamic and basal ganglia hypoperfusion. CONCLUSIONS: We describe rCBF changes in PD in neural circuits related with control of movements. These changes are more manifest in patients with a longer duration of the disease. PMID- 19558952 TI - [Brain perfusion study in patients with systemic lupus erythematosus and neuropsychiatric symptoms]. AB - Neuropsychiatric manifestations in 25% to 70% of patients with systemic lupus erythematosus (SLE), generally in young people. The variability in its clinical expression and lack of diagnostic methods have hindered the diagnosis of Central Nervous System Lupus. When the literature was reviewed on this subject, an important variability was found between the Single Photon Emission Computed Tomography (SPECT) findings and the patient's clinical symptoms and disease course. The case we are presenting shows the usefulness of brain perfusion SPECT because it shows alterations in the central nervous system that are not detected with other imaging modalities. PMID- 19558953 TI - [Contribution of (99m)Tc-mebrofenin scintigraphy to the diagnosis of ectopic liver. A case report]. AB - We present the case of a woman with a parasplenic mass who underwent abdominal ultrasounds, computed tomography and magnetic resonance imaging. The liver scintigraphy with (99m)Tc-Mebrofenin confirmed the diagnosis of ectopic liver. PMID- 19558954 TI - [Bilateral drainage in the internal mammary chain in the detection of the sentinel lymph node in a breast tumor]. PMID- 19558955 TI - [Incidental finding of bone marrow metastatic dissemination in the (99m)Tc-HMPAO leukocyte scintigraphy]. PMID- 19558956 TI - [Somatostatin receptor scintigraphy in a pulmonary carcinoid tumor associated with pneumonia]. PMID- 19558957 TI - [Liver disease: imaging techniques and new therapies]. PMID- 19558958 TI - Potential clinical utility of dual time point FDG-PET for distinguishing benign from malignant lesions: implications for oncological imaging. PMID- 19558959 TI - [Proposal for a diagnostic algorithm for the use of (18)F-FDG-PET in lung cancer]. PMID- 19558960 TI - Identification of a multidrug efflux pump in Flavobacterium johnsoniae. AB - In this study, the mechanism conferring multiple drug resistance in several strains of flavobacteria isolated from the ovarian fluids of hatchery reared 3 year old brook trout Salvelinus fontinalis was investigated. Metabolic fingerprinting and 16S rRNA gene sequences identified the isolates as Flavobacterium johnsoniae. The isolates exhibited multiple resistances to a wide range of antimicrobial classes including penicillin, cephem, monobactam, aminoglycoside, and phenicol. Although plasmids and other transposable elements containing antimicrobial resistance genes were not detected, the isolates did contain a genomic sequence for a chloramphenicol-inducible resistance-nodulation division family multidrug efflux pump system. Efflux pumps are non-specific multidrug efflux systems. They are also a component of cell-cell communication systems, and respond specifically to cell membrane stressors such as oxidative or nitrosative stress. Understanding of efflux pump mediated antibiotic resistances will affect efficacy of clinical treatments of fishes associated with F. johnsoniae epizootics. PMID- 19558961 TI - Reconstruction of the archaeal isoprenoid ether lipid biosynthesis pathway in Escherichia coli through digeranylgeranylglyceryl phosphate. AB - The membrane lipids of archaea are characterized by unique isoprenoid biochemistry, which typically is based on two core lipid structures, sn-2,3 diphytanylglycerol diether (archaeol) and sn-2,3-dibiphytanyldiglycerol tetraether (caldarchaeol). The biosynthetic pathway for the tetraether lipid entails unprecedented head-to-head coupling of isoprenoid intermediates by an unknown mechanism involving unidentified enzymes. To investigate the isoprenoid ether lipid biosynthesis pathway of the hyperthermophilic archaeon, Archaeoglobus fulgidus, its lipid synthesis machinery was reconstructed in an engineered Escherichia coli strain in an effort to demonstrate, for the first time, efficient isoprenoid ether lipid biosynthesis for the production of the intermediate, digeranylgeranylglyceryl phosphate (DGGGP). The biosynthesis of DGGGP was verified using an LC/MS/MS technique and was accomplished by cloning and expressing the native E. coli gene for isopentenyl diphosphate (IPP) isomerase (idi), along with the A. fulgidus genes for G1P dehydrogenase (egsA) and GGPP synthase (gps), under the control of the lac promoter. The A. fulgidus genes for GGGP synthase (GGGPS) and DGGGP synthase (DGGGPS), under the control of the araBAD promoter, were then introduced and expressed to enable DGGGP biosynthesis in vivo. This investigation established roles for four A. fulgidus genes in the isoprenoid ether lipid pathway for DGGGP biosynthesis and provides a platform useful for identification of subsequent, currently unknown, steps in tetraether lipid biosynthesis proceeding from DGGGP, which is the presumed substrate for the head-to-head coupling reaction yielding unsaturated caldarchaeol. PMID- 19558962 TI - Establishment of a novel anabolism-based addiction system with an artificially introduced mevalonate pathway: complete stabilization of plasmids as universal application in white biotechnology. AB - Plasmid stability in recombinant microorganisms is a very important requirement for highly efficient plasmid-based production processes in biotechnology. To stably maintain plasmids, we developed in this study an efficient and stringent novel anabolism-based addiction system, which can be widely used. This novel addiction system is based on two components: (i) an Escherichia coli HMS174(DE3) knockout mutant of the ispH gene coding for 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (EC 1.17.1.2) of the deoxyxylulose 5-phosphate (DXP) pathway, impairing the synthesis of isopentenyl pyrophosphate (IPP) and (ii) a completely synthetic and episomal mevalonate (MVA) pathway as an alternative supplier of essential IPP. The latter is encoded by a plasmid that contains the genes for HMG-CoA reductases from Lactococcus lactis and Staphylococcus aureus plus HMG-CoA-synthase, MVA kinase, MVP kinase and MVPP decarboxylase from S. aureus. This plasmid should then also harbor the genes for the protein or for the pathway that will be produced or that will be utilized for production of a chemical. To demonstrate the functionality of this addiction system, a mutated cyanophycin synthetase gene (cphA(6308)C595S) was used. To determine plasmid stabilities, flasks experiments in media supplied or not supplied with antibiotics were carried out with the knockout mutant and two control strains, one harboring plasmid pCOLADuet-1::MVA1-5::cphA(6308) and the other harboring a conventional expression plasmid pET-23a::cphA(6308). As revealed by measuring the colony-forming units of aliquots spread on solid media with or without antibiotics, the knockout mutant revealed a plasmid stability of 100% whereas the control strains exhibited plasmid stabilities of only 64% and 2%, respectively. Radiometric enzyme activity measurements for CphA revealed only 95% and 12.5% of the activity in the control strains harboring pCOLADuet-1::MVA1-5::cphA(6308) and pET-23a::cphA(6308), respectively, in comparison to the activity measured in the knockout mutant. The knockout mutant synthesized 9.5% (w/w of cell dry weight (CDW)) of cyanophycin, and the control strain harboring pCOLADuet-1::MVA1 5::cphA(6308) synthesized 13.6% (w/w of CDW) after growth without antibiotics. PMID- 19558963 TI - Improving d-mannitol productivity of Escherichia coli: impact of NAD, CO2 and expression of a putative sugar permease from Leuconostoc pseudomesenteroides. AB - The highly productive whole-cell biotransformation of D-fructose to D-mannitol with recombinant, resting cells of Escherichia coli BL21(DE3) requires the combined expression of mdh, fdh and glf which encode mannitol and formate dehydrogenases and a sugar facilitator, respectively. However, long-term stability of the system was restricted, possibly due to loss of the cofactor NAD, high concentrations of formate, formation of CO(2) affecting the internal pH of the cells, accumulation of high intracellular concentrations of D-mannitol, and export of D-mannitol. Downstream of the mdh gene of Leuconostoc pseudomesenteroides, we identified an open reading frame encoding for a putative mannitol permease. The gene was cloned and expressed in E. coli. Biochemical analyses revealed an activity as secondary carrier for D-fructose. Therefore, the carrier was named FupL and participation in D-mannitol transport was excluded. In biotransformation experiments, the productivity of D-mannitol formation obtained with the strain expressing the additional fupL gene was enhanced by 20%. PMID- 19558964 TI - Improving cellular malonyl-CoA level in Escherichia coli via metabolic engineering. AB - Escherichia coli only maintains a small amount of cellular malonyl-CoA, impeding its utility for overproducing natural products such as polyketides and flavonoids. Here, we report the use of various metabolic engineering strategies to redirect the carbon flux inside E. coli to pathways responsible for the generation of malonyl-CoA. Overexpression of acetyl-CoA carboxylase (Acc) resulted in 3-fold increase in cellular malonyl-CoA concentration. More importantly, overexpression of Acc showed a synergistic effect with increased acetyl-CoA availability, which was achieved by deletion of competing pathways leading to the byproducts acetate and ethanol as well as overexpression of an acetate assimilation enzyme. These engineering efforts led to the creation of an E. coli strain with 15-fold elevated cellular malonyl-CoA level. To demonstrate its utility, this engineered E. coli strain was used to produce an important polyketide, phloroglucinol, and showed near 4-fold higher titer compared with wild-type E. coli, despite the toxicity of phloroglucinol to cell growth. This engineered E. coli strain with elevated cellular malonyl-CoA level should be highly useful for improved production of important natural products where the cellular malonyl-CoA level is rate-limiting. PMID- 19558965 TI - Production of flavin mononucleotide by metabolically engineered yeast Candida famata. AB - Recombinant strains of the flavinogenic yeast Candida famata able to overproduce flavin mononucleotide (FMN) that contain FMN1 gene encoding riboflavin (RF) kinase driven by the strong constitutive promoter TEF1 (translation elongation factor 1alpha) were constructed. Transformation of these strains with the additional plasmid containing the FMN1 gene under the TEF1 promoter resulted in the 200-fold increase in the riboflavin kinase activity and 100-fold increase in FMN production as compared to the wild-type strain (last feature was found only in iron-deficient medium). Overexpression of the FMN1 gene in the mutant that has deregulated riboflavin biosynthesis pathway and high level of riboflavin production in iron-sufficient medium led to the 30-fold increase in the riboflavin kinase activity and 400-fold increase in FMN production of the resulted transformants. The obtained C. famata recombinant strains can be used for the further construction of improved FMN overproducers. PMID- 19558966 TI - Hepatic anaplerotic outflow fluxes are redirected from gluconeogenesis to lactate synthesis in patients with Type 1a glycogen storage disease. AB - Hepatic glucose production and relative Krebs cycle fluxes (indexed to a citrate synthase flux of 1.0) were evaluated with [U-(13)C]glycerol tracer in 5 fed healthy controls and 5 Type 1a glycogen storage disease (GSD1a) patients. Plasma glucose, hepatic glucose-6-phosphate (G6P) and glutamine (13)C-isotopomers were analyzed by (13)C NMR via blood sampling and chemical biopsy. In healthy subjects, 35+/-14% of plasma glucose originated from hepatic G6P while GSD1a patients had no detectable G6P contribution. Compared to controls, GSD1a patients had an increased fraction of acetyl-CoA from pyruvate (0.5+/-0.2 vs. 0.3+/-0.1, p<0.01), and increased pyruvate recycling fluxes (14.4+/-3.8 vs. 8.7+/-2.8, p<0.05). Despite negligible gluconeogenic flux, net anaplerotic outflow was not significantly different from controls (2.2+/-0.8 vs. 1.3+/-0.5). The enrichment of lactate with (13)C-isotopomers derived from the Krebs cycle suggests that lactate was the main anaplerotic product in GSD1a patients. PMID- 19558967 TI - Construction of a synthetic YdbK-dependent pyruvate:H2 pathway in Escherichia coli BL21(DE3). AB - A synthetic pyruvate:H(2) pathway was constructed in Escherichia coli BL21(DE3) by co-expression of six proteins: E. coli YdbK, Clostridium pasteurianum [4Fe-4S] ferredoxin, and Clostridium acetobutylicum HydF, HydE, HydG, and HydA. The effect of cofactor addition and host strain on H(2) yield and fermentation product accumulation was studied, together with in vitro reconstitution of the entire pathway. The deletion of iscR and/or the addition of thiamine pyrophosphate to the medium enhanced the total and specific activity of recombinant YdbK and increased the yield of H(2) per glucose. It was concluded that the introduced pathway outcompeted other pyruvate-consuming reactions, and that the ability to compete for pyruvate at least in part was determined by total YdbK activity. The results demonstrate the successful construction of a high-yielding H(2) pathway in a microorganism that effectively does not synthesize any H(2). The additional co-expression of Bacillus subtilis AmyE enabled starch-dependent H(2) synthesis in minimal media. PMID- 19558973 TI - Peripheral nerve at extreme low temperatures 2: pharmacologic modulation of temperature effects. AB - Changes in temperature have profound and clinically important effects on the peripheral nerve. In a previous paper, the effects of temperature on many properties of the peripheral nerve action potential (NAP) were explored including the NAP amplitude, conduction velocity and response to paired pulse stimulation. In this paper, the effects of pharmacologic manipulations on these parameters were explored in order to further understand the mechanisms of these effects. The reduction in conduction velocity with temperature was shown to be independent of the ionic composition of the perfusate and was unaffected by potassium or sodium channel blockade. This implies that the phenomenon of reduced conduction velocities at low temperature may be related to changes in the passive properties of the axon with temperature. Blockade of sodium channels and chronic membrane depolarization produced by high perfusate potassium concentrations or high dose 4 aminopyridine impair the resistance of the nerve to hypothermia and enhance the injury to the nerve produced by cycles of cooling and rewarming. This suggests the possibility that changes in the sodium inactivation channel may be responsible for the changes in the NAP amplitude with temperature and that prolonged sodium inactivation may lead more permanent changes in excitability. PMID- 19558975 TI - Peripheral nerve at extreme low temperatures 1: effects of temperature on the action potential. AB - Hypothermia is an important means of neuroprotection. Understanding the effects of temperature on a physiologic measurement such as the nerve action potential (NAP) is important in monitoring its effects. The effects of hypothermia on the NAP amplitude, conduction velocity, and response to paired pulse stimulation were quantified in a rat sciatic nerve preparation from 37 to 10 degrees C. The time course of temperature related changes and the effect of repeated cycles of cooling and rewarming are explored using the following measures of the NAP: peak to-peak amplitude, conduction velocity, duration, area under the curve and response to paired pulse stimuli. The NAP amplitude initially increases as temperature is reduced to 27 degrees C and then drops to roughly 50% of its baseline value by 16 degrees C while the area under the curve increases gradually until it begins to decline at 16 degrees C. Permanent loss of the NAP appears only after cooling below 10 degrees C for extended periods. Although the dependence of amplitude on temperature is approximately sigmoidal, the conduction velocity declines linearly at a rate of 2.8m/s/ degrees C. The response to paired pulse stimulation is strongly dependent on both temperature and the interstimulus interval with the responses at shorter interstimulus intervals being more temperature sensitive. With repetitive cycles of cooling and rewarming, the NAP amplitude declines by roughly 4% with every cycle without changes in the temperature at which the NAP amplitude reaches 50% of baseline. Only minor differences in conduction velocity are seen during cooling and rewarming. PMID- 19558979 TI - Ruptured hepatoma with hemorrhagic ascites. PMID- 19558984 TI - An unusual cause of hematemesis. PMID- 19558985 TI - Hepatic portal venous gas caused by emphysematous pyelonephritis. PMID- 19558986 TI - Gastric heterotopia in the rectum. PMID- 19558987 TI - Clinical scenario--an 18-year-old with acute dysphagia and meat impaction. PMID- 19558989 TI - Analysis of court criteria for awarding disability benefits to patients with Crohn's disease. AB - BACKGROUND & AIMS: Chronic disability and its consequences for social life and employment are important but often neglected aspects of Crohn's disease. No specific scores have been developed to evaluate chronic disability in patients with Crohn's disease; the medical criteria used by government authorities to award disability benefits have not been analyzed. We aimed to determine the courts' criteria for awarding disability benefits to patients with Crohn's disease in Spain. METHODS: We systematically searched case law databases in Spain's regional Supreme Courts to identify sentences regarding awards of disability benefits to patients with Crohn's disease. Selected decisions were reviewed to extract variables related to the awarding of benefits. Univariate and multivariate analyses were performed to determine which variables predicted the awarding of benefits. RESULTS: Two hundred eighty sentences were reviewed. The rate of judicial decisions in favor of the claimants varied considerably between the various tribunals. Multivariate analysis showed that adequate description of the disease (odds ratio, 8.6), fecal incontinence (odds ratio, 8.9), the number of associated diseases (odds ratio, 2.3), and the presence of an ostomy (odds ratio, not estimable) were independent predictors of the awarding of Social Security benefits. CONCLUSIONS: The amount of Social Security benefits awarded to patients with Crohn's disease varied depending on the tribunal. The most important predictors of a court's disability award were the adequate description of the patient's disease, fecal incontinence, associated diseases, and presence of an ostomy. PMID- 19558992 TI - Hepatic artery pseudoaneurysm associated with cholangitis that ruptured into the bile duct. PMID- 19558993 TI - Peutz-Jeghers syndrome: a rare cause of gastric outlet obstruction. PMID- 19558995 TI - Iron ulcers. PMID- 19558997 TI - Risk of lymphoma associated with combination anti-tumor necrosis factor and immunomodulator therapy for the treatment of Crohn's disease: a meta-analysis. AB - BACKGROUND & AIMS: Although anti-tumor necrosis factor (TNF) therapy can effectively treat Crohn's disease (CD), there is concern that it might increase the risk of non-Hodgkin's lymphoma (NHL). A meta-analysis was performed to determine the rate of NHL in adult CD patients who have received anti-TNF therapy and to compare this rate with that of a population-based registry and a population of CD patients treated with immunomodulators. METHODS: MEDLINE, EMBASE, Cochrane Collaboration, and Web of Science were searched. Inclusion criteria included randomized controlled trials, cohort studies, or case series reporting on anti-TNF therapy in adult CD patients. Standardized incidence ratios (SIR) were calculated by comparing the pooled rate of NHL with the expected rate of NHL derived from the Surveillance Epidemiology & End Results (SEER) database and a meta-analysis of CD patients treated with immunomodulators. RESULTS: Twenty six studies involving 8905 patients and 21,178 patient-years of follow-up were included. Among anti-TNF treated subjects, 13 cases of NHL were reported (6.1 per 10,000 patient-years). The majority of these patients had previous immunomodulator exposure. Compared with the expected rate of NHL in the SEER database (1.9 per 10,000 patient-years), anti-TNF treated subjects had a significantly elevated risk (SIR, 3.23; 95% confidence interval, 1.5-6.9). When compared with the NHL rate in CD patients treated with immunomodulators alone (4 per 10,000 patient-years), the SIR was 1.7 (95% confidence interval, 0.5-7.1). CONCLUSIONS: The use of anti-TNF agents with immunomodulators is associated with an increased risk of NHL in adult CD patients, but the absolute rate of these events remains low and should be weighed against the substantial benefits associated with treatment. PMID- 19558999 TI - Effect of chronic hepatitis C virus infection on bone disease in postmenopausal women. AB - BACKGROUND & AIMS: Limited data are available on the contribution of chronic HCV infection to the development of bone disease in postmenopausal women. We studied whether women who acquired HCV infection through administration of HCV genotype 1b-contaminated anti-D immunoglobulin from a single source had decreased bone mineral density (BMD) or altered levels of bone turnover markers (BTMs), compared with women who spontaneously resolved infection or age-matched healthy controls. METHODS: From a cohort of postmenopausal Irish women, we compared BMD, determined by dual-energy x-ray absorptiometry, and a panel of BTMs in 20 women chronically infected with HCV (PCR(+)), 21 women who had spontaneously resolved infection (PCR(-)), and 23 age-matched healthy controls. RESULTS: Levels of BTMs and BMD were similar in PCR(+) and PCR(-) women and healthy age-matched controls. However, there was an increased frequency of fractures in PCR(+) (n = 6) compared with PCR(-) women (n = 0, P = .007). PCR(+) women with fractures were postmenopausal for a longer time (median, 15.5, range, 5-20 years vs 4.5, range, 1-20 years in PCR(+) women without fractures; P = .033), had lower BMD at the hip (0.79, range, 0.77-0.9 g/cm(2) vs 0.96, range, 0.81-1.10 g/cm(2); P = .007), and had a lower body mass index (23.7, range 21.2-28.5 kg/m(2) vs 25.6, range 22.1 36.6 kg/m(2); P = .035). There was no difference in liver disease severity or BTMs in PCR(+) women with or without fractures. CONCLUSIONS: Chronic HCV infection did not lead to discernable metabolic bone disease in postmenopausal women, but it might be a risk factor for bone fractures, so preventive measures should be introduced. To view this article's video abstract, go to the AGA's YouTube Channel. PMID- 19559002 TI - The suitability of clay pots for indoor sampling of mosquitoes in an arid area in northern Tanzania: A word of caution. PMID- 19559003 TI - The effect of salt and pH on block liposomes studied by cryogenic transmission electron microscopy. AB - Recently, we have reported the discovery of block liposomes (BLs), a new class of liquid (chain-melted) vesicles, formed in mixtures of the curvature-stabilizing hexadecavalent cationic lipid MVLBG2, the neutral lipid 1,2-dioleoyl-sn-glycero-3 phosphatidylcholine (DOPC), and water with no added salt. BLs consist of connected spheres, pears, tubes, or rods. Unlike in typical liposome systems, where spherical vesicles, tubular vesicles, and cylindrical micelles are separated on the macroscopic scale, shapes remain connected and are separated only on the nanometer scale within a single BL. Here, we report structural studies of the effect of salt and pH on the BL phase, carried out using differential interference contrast microscopy (DIC) and cryogenic transmission electron microscopy (cryo-TEM). Addition of salt screens the electrostatic interactions; in low-salt conditions, partial screening of electrostatic interactions leads to a shape transition from BLs to bilamellar vesicles, while in the high-salt regime, a shape transition from BLs to liposomes with spherical morphologies occurs. This demonstrates that strong electrostatic interactions are essential for BL formation. Understanding the control of liposome shape evolution is of high interest because such shape changes play an important role in many intracellular processes such as endocytosis, endoplasmatic reticulum-associated vesiculation, vesicle recycling and signaling. PMID- 19559004 TI - Characterization of the mechanism of action of the genetically modified Cry1AbMod toxin that is active against Cry1Ab-resistant insects. AB - Bacillus thuringiensis Cry toxins are used in the control of insect pests. They are pore-forming toxins with a complex mechanism that involves the sequential interaction with receptors. They are produced as protoxins, which are activated by midgut proteases. Activated toxin binds to cadherin receptor, inducing an extra cleavage including helix alpha-1, facilitating the formation of a pre-pore oligomer. The toxin oligomer binds to secondary receptors such as aminopeptidase and inserts into lipid rafts forming pores and causing larval death. The primary threat to efficacy of Bt-toxins is the evolution of insect resistance. Engineered Cry1AMod toxins, devoid of helix alpha-1, could be used for the control of resistance in lepidopterans by bypassing the altered cadherin receptor, killing resistant insects affected in this receptor. Here we analyzed the mechanism of action of Cry1AbMod. We found that alkaline pH and the presence of membrane lipids facilitates the oligomerization of Cry1AbMod. In addition, tryptophan fluorescence emission spectra, ELISA binding to pure aminopeptidase receptor, calcein release assay and analysis of ionic-conductance in planar lipid bilayers, indicated that the secondary steps in mode of action that take place after interaction with cadherin receptor such as oligomerization, receptor binding and pore formation are similar in the Cry1AbMod and in the wild type Cry1Ab. Finally, the membrane-associated structure of Cry1AbMod oligomer was analyzed by electron crystallography showing that it forms a complex with a trimeric organization. PMID- 19559005 TI - Measurement of (5'R)- and (5'S)-8,5'-cyclo-2'-deoxyadenosines in DNA in vivo by liquid chromatography/isotope-dilution tandem mass spectrometry. AB - Oxidatively induced DNA lesions (5'R)- and (5'S)-8,5'-cyclo-2'-deoxyadenosines (R cdA and S-cdA) are detectable and accumulate in vivo due to disease states and defects in DNA repair. They block transcription and inhibit gene expression, and may play a role in disease processes. Accurate measurement of these lesions in DNA in vivo is necessary to understand their biological effects. We report on a methodology using liquid chromatography/isotope-dilution tandem mass spectrometry to measure R-cdA and S-cdA in DNA. This methodology permitted the detection of these compounds at a level of 0.1fmol on-column. Levels of R-cdA and S-cdA in mouse liver DNA amounted to 0.133+/-0.024 and 0.498+/-0.065 molecules/10(7) DNA 2'-deoxynucleosides, respectively. The successful measurement of R-cdA and S-cdA in DNA in vivo suggests that this methodology will be used for understanding of their repair and biological consequences, and that these compounds may be used as putative biomarkers for disease states. PMID- 19559006 TI - A major mutation of KIF21A associated with congenital fibrosis of the extraocular muscles type 1 (CFEOM1) enhances translocation of Kank1 to the membrane. AB - Congenital fibrosis of the extraocular muscles type 1 (CFEOM1) is associated with heterozygous mutations in the KIF21A gene, including a major (R954W) and a minor (M947T) mutation. Kank1, which regulates actin polymerization, cell migration and neurite outgrowth, interacted with the third and fourth coiled-coil domains of KIF21A protein at its ankyrin-repeat domain. While both KIF21A(R954W) and KIF21A(M947T) enhanced the formation of a heterodimer with the wild type, KIF21A(WT), these mutants also enhanced the interaction with Kank1. Knockdown of KIF21A resulted in Kank1 predominantly occurring in the cytosolic fraction, while KIF21A(WT) slightly enhanced the translocation of Kank1 to the membrane fraction. Moreover, KIF21A(R954W) significantly enhanced the translocation of Kank1 to the membrane fraction. These results suggest that KIF21A regulates the distribution of Kank1 and that KIF21A mutations associated with CFEOM1 enhanced the accumulation of Kank1 in the membrane fraction. This might cause an abrogation of neuronal development in cases of CFEOM1 through over-regulation of actin polymerization by Kank1. PMID- 19559007 TI - Attenuation of LPS-induced iNOS expression by 1,5-anhydro-d-fructose. AB - 1,5-anhydro-d-fructose (1,5-AF), a monosaccharide formed from starch and glycogen, exhibits antioxidant and antibacterial activity, and inhibits cytokine release by attenuating NF-kappaB activation in LPS-stimulated mice. The present study examined whether 1,5-AF inhibits lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) in vitro and in vivo. We found that 1,5-AF significantly blocked the production of NO, and protein and mRNA expression of iNOS, and up-regulated IL-10 production in vitro. We also investigated the effects of 1,5-AF on acute lung inflammation in C57BL/6J mice. We found that protein and mRNA expression of iNOS in lung tissues were inhibited by 1,5-AF pretreatment. In addition, the serum level of IL-10 was upregulated by 1,5-AF. Collectively, the iNOS transcriptional and translational inhibitory effects of 1,5-AF seem to be prolonged and enhanced by the production of IL-10. These results suggest that 1,5-AF could be a useful adjunct in the treatment of acute lung inflammation. PMID- 19559008 TI - Synthesis of fluorescent-maghemite nanoparticles as multimodal imaging agents for amyloid-beta fibrils detection and removal by a magnetic field. AB - Early diagnosis in Alzheimer's disease (AD), before the onset of marked clinical symptoms, is critical in preventing the irreversible neuronal damage that eventually leads to dementia and ultimately death. Therefore, there is an urgent need for in vivo imaging agents, which are valuable as specific biomarkers to demonstrate the location and density of amyloid plaques in the living human brain. The present manuscript describes a novel method for selective marking of Abeta(40) fibrils by non-fluorescent gamma-Fe(2)O(3) and fluorescent-magnetic gamma-Fe(2)O(3)-rhodamine or gamma-Fe(2)O(3)-Congo red nanoparticles, and the complete removal of the magnetized fibrils from the aqueous continuous phase by a magnetic field. These fluorescent-maghemite nanoparticles as multimodal imaging agents have a great advantage due to the combination of the magnetic and fluorescence imaging into one nanostructured system. This hybrid system, which selectively marks Abeta(40) fibrils, might enable the early detection of plaques using both MRI and fluorescence microscopy, and therefore may be applied in in vivo AD diagnosis studies. These fluorescent-magnetic nanoparticles may also be useful as selective biomarkers to detect the location and the removal of other amyloid plaques derived from different amyloidogenic proteins that lead to neurodegenerative diseases, e.g., Parkinson's, Huntington's, mad cow, and prion diseases. PMID- 19559009 TI - Cytosolic labile zinc accumulation in degenerating dopaminergic neurons of mouse brain after MPTP treatment. AB - High levels of labile zinc accumulate in degenerating neurons after brain injury, such as ischemic stroke, trauma, epilepsy, and hypoglycemia. Cytosolic zinc accumulation is also found in brain neurons undergoing apoptosis during development or after neuronal target ablation. Thus, staining with zinc-specific probes can be used to identify neuronal death in the brain. In this study, mice were intraperitoneally given four 20 mg/kg doses of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) at 2-hour intervals, and dopaminergic neurons were then evaluated for zinc accumulation and apoptosis. In the substantia nigra pars compacta, zinc-specific fluorescent dyes revealed that all degenerating neurons, identified by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL), or acid fuchsin or Fluoro-Jade C staining, contained high levels of cytosolic labile zinc. Nuclear condensation/fragmentation was noted in dopaminergic neurons with cytosolic zinc accumulation, indicating apoptotic cell death. These findings support the supposition that cytosolic labile zinc accumulation is an indicator of degenerating dopaminergic neurons in animal models of Parkinson's disease. PMID- 19559010 TI - Abnormalities of the uncinate fasciculus and posterior cingulate fasciculus in mild cognitive impairment and early Alzheimer's disease: a diffusion tensor tractography study. AB - Mild cognitive impairment (MCI) is considered the transitional stage between normal cognition and dementia. The aim of this study was to use tractography based analysis to elucidate alterations in subjects with MCI compared with subjects with early Alzheimer's disease (AD) and controls. Seventeen subjects with early AD, 16 with MCI and 16 controls underwent magnetic resonance diffusion tensor imaging (DTI) and neuropsychological assessment. Diffusion tensor tractographies were computed and fiber-tract maps were generated using "dTV II" DTI software. We measured mean fractional anisotropy (FA) and apparent diffusion coefficient (ADC) values along the uncinate fasciculus (UNC), posterior cingulate fasciculus (PCF) and corticospinal tract (CST). There were statistically significant differences in the FA and ADC values of the UNC and PCF between subjects with early AD and controls. Subjects with MCI exhibited significantly lower FA values on both sides of the PCF relative to controls. However, there were no significant differences between subjects with early AD and MCI for any measurement. Our results suggest that alterations in the PCF precede the onset of dementia. PMID- 19559012 TI - The impact of paracrine signaling in brain microvascular endothelial cells on the survival of neurons. AB - Neurons depend for survival on local neurotrophic factors which act in an autocrine/paracrine manner. However, the effect of paracrine signaling of brain microvascular endothelial cells (BMECs) under pathological conditions on neuron survival is not fully understood. In this study we cultured rat BMECs and cortical neurons. BMECs were cultured in oxygen-glucose-deprived (OGD) conditions to mimic cerebral ischemia in vitro. The conditioned media of normal BMECs or OGD injured BMECs were used to culture normal or injured neurons. Neuron activity, free Ca(2+) concentration, NMDA receptor status, mitochondrial membrane potential and cytochrome C release level were determined. The results showed: mitochondrial activity of injured neurons was significantly increased and lactate dehydrogenase (LDH) leakage was decreased (P<0.05) by grown in conditioned medium of normal BMECs. Inversely, mitochondrial activity of normal or injured neurons was decreased and LDH leakage was significantly increased (P<0.05) by grown in conditioned medium of injured BMECs. The changes in free Ca(2+) concentration, NMDA receptor status, mitochondrial membrane potential and cytochrome C release level were consistent with the changes in neuronal activity. These findings suggest that the conditioned medium of normal BMECs has a neuroprotective effect. However, this protective effect was lost after BMECs injury; in fact, the conditioned medium became neurotoxic. Therefore, it appears that early recovery of BMECs might be helpful for neuron survival. PMID- 19559011 TI - ATF3 plays a protective role against toxicity by N-terminal fragment of mutant huntingtin in stable PC12 cell line. AB - Huntington's disease is a progressive neurodegenerative disorder caused by a polyglutamine expansion near the N-terminus of huntingtin. The mechanisms of polyglutamine neurotoxicity, and cellular responses are not fully understood. We have studied gene expression profiles by short oligo array using an inducible PC12 cell model expressing an N-terminal huntingtin fragment with expanded polyglutamine (Htt-N63-148Q). Mutant huntingtin Htt-N63 induced cell death and increased the mRNA and protein levels of activating transcription factor 3 (ATF3). Mutant Htt-N63 also significantly enhanced ATF3 transcriptional activity by a promoter-based reporter assay. Overexpression of ATF3 protects against mutant Htt-N63 toxicity and knocking down ATF3 expression reduced Htt-N63 toxicity in a stable PC12 cell line. These results indicated that ATF3 plays a critical role in toxicity induced by mutant Htt-N63 and may lead to a useful therapeutic target. PMID- 19559013 TI - Transcriptional induction and translational inhibition of Arc and Cugbp2 in mice hippocampus after transient global ischemia under normothermic condition. AB - Mild hypothermia protects against neuronal damage after transient global ischemia in experimental animals. The exact mechanism of this protective effect remains to be elucidated. The purpose of the present study was to investigate the molecular mechanisms relevant to different neurologic responses to hypothermia and normothermia. Transient global ischemia was induced in C57BL/6 mice by bilateral common carotid artery occlusion for 10 min. Hypothermia provided robust neuroprotection in the hippocampus region and dramatically reduced the mortality rate. Using adaptor-tagged competitive polymerase chain reaction, we obtained the relative transcription levels of 1210 genes in the hippocampal region and compared the expression patterns of these genes. Two genes, Activity-regulated cytoskeleton-associated protein (Arc) and CUG-binding protein-2 (Cugbp2), showed remarkable and persistent increases in their expression levels in normothermic mice, compared with in both sham and hypothermic mice. Despite the increased transcription of Arc and Cugbp2, an immunohistochemistry analysis did not show comparable increases in the translations of both genes. Only a transient increase in Arc protein was observed in the granule cells of the dentate gyrus at 6 h after reperfusion. A remarkable decrease in Cugbp2 protein was observed in the pyramidal cells of the hippocampal CA1-CA3, in accordance with the progress of neuronal degeneration. A decrease in Cugbp2 protein was not observed in hypothermic mice. These results suggest that transient global ischemia induces the translational inhibition of genes with increased expression not in hypothermic, but in normothermic mice. Thus, translational inhibition might play an important role in the progress of neuronal injury after transient global ischemia. PMID- 19559014 TI - rPer1 and rPer2 induction during phases of the circadian cycle critical for light resetting of the circadian clock. AB - Photic resetting of a biological clock is one of the fundamental characteristics of circadian systems and allows living organisms to adjust to a particular environment. Nocturnal light induces the Per1 and Per2 genes, which leads to a resetting of the circadian clock in the suprachiasmatic nucleus (SCN), the mammalian circadian center. In our present study, we investigated whether light differentially induces the rat Per1 (rPer1) and Per2 (rPer2) genes to enable resetting of their circadian clocks. In a 24-hour LD cycle (12 h light:12 h dark), which is shorter than the normal free-running period for rats, Per1 alone showed strong induction in the ventrolateral region of the SCN (VLSCN) during the early day. In contrast, in a 25 hour LD cycle (12.5 h light:12.5 h dark), which is longer than the free running period for these animals, rPer2 alone was strongly induced in the VLSCN, at the end of the light phase and during the early dark periods. Our current findings therefore suggest that Per1 and Per2 are differentially regulated for daily entrainment to the LD cycle. PMID- 19559015 TI - MicroRNA-21 targets LRRFIP1 and contributes to VM-26 resistance in glioblastoma multiforme. AB - MicroRNAs control a wide array of biological processes including cell differentiation, proliferation, and apoptosis whose dysregulation is a hallmark of cancer. MicroRNA-21 (miR-21) is overexpressed in many cancers including glioblastoma and contributes to tumor resistance to chemotherapy. We investigated whether miR-21 mediated chemoresistance to the chemotherapeutic agent VM-26 in glioblastoma cells and sought to identify the candidate target genes for miR-21 by gene expression profiling. Here we report that miR-21 was involved in mediating chemoresistance to VM-26 in glioblastoma cells. Suppression of miR-21 by specific antisense oligonucleotides in glioblastoma cell U373 MG led to enhanced cytotoxicities of VM-26 against U373 MG cells. We further identified and validated LRRFIP1, whose product is an inhibitor of NF-kappaB signaling, as a direct target gene of miR-21. Our findings suggest that miR-21 represents a promising target for therapeutic manipulation to increase the efficacy of chemotherapeutic agents in treating glioblastoma, a highly lethal type of cancer. PMID- 19559016 TI - Effect of dimethyl sulfoxide on the phase behavior of model stratum corneum lipid mixtures. AB - Dimethyl sulfoxide (DMSO), an efficient transdermal enhancer, is proposed to alter the skin barrier by, at least partially, disturbing the lipid phase of the stratum corneum (SC). We have investigated, using differential scanning calorimetry and vibrational microspectroscopy, the effect of DMSO on the phase behavior of a lipid mixture formed by N-palmitoyl-d-erythro-sphingosine, deuterated palmitic acid, and cholesterol, mimicking the SC lipid phase. Our results reveal that DMSO favors the disordering of the lipid acyl chains. Moreover, the effect of DMSO is strongly concentration dependent and this dependence is reminiscent of that describing the DMSO transdermal enhancement. DMSO-induced fluidification affects primarily the fatty acid in the mixture. Therefore, it is proposed that the molecular mechanism of the transdermal transport enhancement caused by DMSO is associated with its H-bonding properties; its presence alters the interfacial H-bond network involving the fatty acid molecules and consequently the cohesive lipid packing. PMID- 19559017 TI - Caucasian-specific allele in non-synonymous single nucleotide polymorphisms of the gene encoding deoxyribonuclease I-like 3, potentially relevant to autoimmunity, produces an inactive enzyme. AB - BACKGROUND: Deoxyribonuclease I-like 3 (DNase Il3), a member of human DNase I family, is postulated to be involved in the genesis of autoimmune diseases. In the DNase Il3 gene, 2 non-synonymous SNPs, R178H and R206C, have been identified, however relevant population data are not available. METHODS: Genotyping of the SNPs was performed in healthy subjects belonging to 3 ethnic groups (n=1708), including nine different populations, using an amplification refractory mutation system and the PCR-RFLP technique. RESULTS: All of the 9 populations were typed as a single genotype in R178H. All Asian and African populations exhibited only a homozygous C686 allele in R206C, whereas a heterozygote, C686/T686, was found (frequency of 3.5-15.4%) in three Caucasian populations (Turk, German and Mexican); Caucasian-specific allele T686 was identified. The substitution of Arg by Cys corresponding to R206C resulted in elimination of DNase Il3 activity. CONCLUSION: A Caucasian-specific allele in SNP R206C produces an inactive form of DNase Il3. It seems plausible that levels of DNase Il3 activity in Caucasian subjects with the heterozygote in R206C are lower than those in individuals with the predominant homozygote. Our results may have clinical implications in relation to the prevalence of autoimmune diseases. PMID- 19559018 TI - Early predictors of microvascular complications in type 1 diabetic patients. AB - OBJECTIVES: To investigate the possibility of depending on adiponectin and leptin as early predictors of microvascular complications in type 1 diabetic subjects. DESIGN AND METHODS: We studied 63 type 1 diabetic subjects from the National Institute of Diabetes (30 normoalbuminuric and 33 microalbuminuric). Clinical, demographic characteristics and kidney function tests were monitored. Plasma levels of adiponectin, leptin, interlukein-6 (IL-6), and the high sensitive C reactive protein (CRP) were measured in these subjects. RESULTS: Microalbuminuric subjects showed a significant elevation in adiponectin levels and a significant decrease in leptin levels as compared to normoalbuminuric subjects. Adiponectin showed a significant positive correlation with microalbuminuria concentrations while leptin showed a significant negative correlation with both fasting blood glucose and glycated hemoglobin A(1c). CONCLUSION: The results of this study introduced the possibility of depending on adiponectin and leptin as early, reliable, and sensitive predictors for the microvascular complications monitored by microalbuminuria concentration and glycemic control indices. PMID- 19559019 TI - Characterization of the adrenomedullin receptor acting as the target of a new radiopharmaceutical biomolecule for lung imaging. AB - Direct labeling of linear adrenomedullin (AM) with (99m)Tc ([(99m)Tc]AM) displayed excellent selectivity for imaging the pulmonary circulation system in dogs. Hence, we investigated this particular selectivity and characterized the binding sites found in dog lungs. AM and other peptides belonging to the calcitonin peptide family, including calcitonin-gene related peptide (CGRP), adrenomedullin-2 (AM2), amylin and pro-adrenomedullin N-terminal peptide (PAMP), were prepared by solid-phase peptide synthesis. Receptor binding assays were performed by using [(125)I]AM as a radioligand on dog lung homogenates. It was found that AM bound with potent affinity, displaying in fact a high and a low affinity binding site. Moreover, competition binding assays using peptide ligands showed the following ranking for displacement: AM>AM(13-52)>CGRP approximately AM2> or =AM(22-52)> or =AM2(16-47)>CGRP(8-37)>amylin approximately PAMP. Thus, these results strongly suggested that the AM binding site found in dog lungs and acting as a clearance receptor is mainly the adrenomedullin AM(1) receptor subtype. The pharmacophores underlying AM(1) binding affinity and specificity were studied by determining the key amino acids, the minimal peptide fragment, and some aspects of the secondary structures. So far, it appeared that the C terminal segment of human AM is an essential feature for binding. Also, the alpha helix secondary structure found in the AM molecule would facilitate the ligand recognition process with the AM receptor in dog lungs. Our results demonstrated that AM or some analogs or fragments could be suitable radiopharmaceutical agents for lung imaging. PMID- 19559020 TI - A reduction of epidermal growth factor receptor is involved in brefelamide induced inhibition of phosphorylation of ERK in human astrocytoma cells. AB - Brefelamide is an aromatic amide isolated from Dictyostelium cellular slime molds. We found that brefelamide has a potent inhibitory growth effect measured by MTT assay in 1321N1 human astrocytoma cells. The inhibition was associated with reduced phosphorylation of extracellular signal-regulated kinase (ERK). Brefelamide inhibited epidermal growth factor (EGF)-induced phosphorylation of ERK in a concentration-dependent manner. Furthermore, brefelamide diminished EGF induced phosphorylation of EGF receptor at Tyr(1068), a Grb2 binding site that leads to an activation of the Ras/Raf/ERK system. Brefelamide also reduced the expression level of the EGF receptor. These results suggest that one of the mechanisms of action of brefelamide is assumed to be inhibition of phosphorylation of ERK through a reduction of EGF receptor activity in 1321N1 human astrocytoma cells. PMID- 19559021 TI - Characterization of the role of full-length CRMP3 and its calpain-cleaved product in inhibiting microtubule polymerization and neurite outgrowth. AB - Collapsin response mediator proteins (CRMPs) are key modulators of cytoskeletons during neurite outgrowth in response to chemorepulsive guidance molecules. However, their roles in adult injured neurons are not well understood. We previously demonstrated that CRMP3 underwent calcium-dependent N-terminal protein cleavage during excitotoxicity-induced neurite retraction and neuronal death. Here, we report findings that the full-length CRMP3 inhibits tubulin polymerization and neurite outgrowth in cultured mature cerebellar granule neurons, while the N-terminal truncated CRMP3 underwent nuclear translocation and caused a significant nuclear condensation. The N-terminal truncated CRMP3 underwent nuclear translocation through nuclear pores. Nuclear protein pull-down assay and mass spectrometry analysis showed that the N-terminal truncated CRMP3 was associated with nuclear vimentin. In fact, nuclear-localized CRMP3 co localized with vimentin during glutamate-induced excitotoxicity. However, the association between the truncated CRMP3 and vimentin was not critical for nuclear condensation and neurite outgrowth since over-expression of truncated CRMP3 in vimentin null neurons did not alleviate nuclear condensation and neurite outgrowth inhibition. Together, these studies showed CRMP3's role in attenuating neurite outgrowth possibility through inhibiting microtubule polymerization, and also revealed its novel association with vimentin during nuclear condensation prior to neuronal death. PMID- 19559022 TI - Angiostrongylus cantonensis: effect of combination therapy with albendazole and dexamethasone on Th cytokine gene expression in PBMC from patients with eosinophilic meningitis. AB - To determine how combination therapy with albendazole and dexamethasone changed cytokine responses in peripheral blood mononuclear cells (PBMC) in patients with eosinophilic meningitis caused by Angiostrongylus cantonensis (EOMA), we measured mRNA levels of Th2 (IL-5, IL-4 and IL-10) and Th1 (IL-2 and IFN-gamma) cytokines with reverse transcription polymerase chain reaction (RT-PCR). Forty-three patients were divided into three groups: group 1 (pre-treatment, 13 patients), group 2 (7 days post-treatment, 14 patients), and group 3 (30 days post treatment, 16 patients). Peripheral eosinophil counts were also measured. EOMA patients showed higher levels of Th2 cytokines, including IL-5 and IL-10, and peripheral eosinophil counts, but no changes in IL-4 or Th1 cytokines. Combination therapy reduced IL-5 mRNA expression and peripheral eosinophil counts to control levels, but increased IL-10, IL-2, and IFN-gamma mRNA expression, and did not change IL-4 levels. These data suggest that systemic Th2 cytokine responses, especially IL-5, and peripheral eosinophil counts increased in EOMA patients. Combination therapy with albendazole and dexamethasone can shift the cytokine responses from Th2 to Th1 dominance, which may be a therapeutic mechanism. PMID- 19559023 TI - Echinococcus multilocularis: Proteomic analysis of the protoscoleces by two dimensional electrophoresis and mass spectrometry. AB - Echinococcus multilocularis is an important parasite that causes human alveolar echinococcosis. Identification and characterization of the proteins encoded by E. multilocularis metacestode might help to understand the complexity of the parasites and their interactions with the host, and to identify new candidates for immunodiagnosis and vaccine development. Here we present a proteomic analysis of E. multilocularis protoscolex (PSC) proteins. The proteins were resolved by 2 DE (pH range 3.5-10), followed by MALDI-TOF MS analysis. Fourteen known Echinococcus proteins were identified, including cytoskeletal proteins, heat shock proteins, metabolic enzymes, 14-3-3 protein, antigen P-29 and calreticulin. To construct a systematic reference map of the immunogenic proteins from E. multilocularis PSC, immunoblot analysis of PSC 2-DE maps was performed. Over 50 proteins spots were detected on immunoblots as antigens and 15 of them were defined. The results showed that cytoskeletal proteins and heat shock proteins were immunodominant antigens in alveolar echinococcosis. PMID- 19559024 TI - alphaB-crystallin extracellularly suppresses ADP-induced granule secretion from human platelets. AB - alphaB-crystallin, a low-molecular-weight heat shock protein (HSP), has binding sites on platelets. However, the exact role of alphaB-crystallin is not clarified. In this study, we investigated the effect of alphaB-crystallin on platelet granule secretion. alphaB-crystallin attenuated the adenosine diphosphate (ADP)-induced phosphorylation of p44/p42 mitogen-activated protein kinase (MAPK) and p38 MAPK. The ADP-stimulated HSP27 phosphorylation was markedly reduced by alphaB-crystallin. alphaB-crystallin significantly suppressed the ADP induced secretions of both platelet-derived growth factor (PDGF)-AB and serotonin. Therefore, our results strongly suggest that alphaB-crystallin extracellularly suppresses platelet granule secretion by inhibition of HSP27 phosphorylation via p44/p42 MAPK and p38 MAPK. PMID- 19559025 TI - Elastin peptides antagonize ceramide-induced apoptosis. AB - Elastin peptides regulate proliferation, chemotaxis and protease expression. The aim of this work was to assess their influence on apoptosis. Human skin fibroblast cell death was induced using C(2)-ceramide in the presence or absence of elastin peptides. We show that ceramide-induced apoptosis could be blocked by elastin peptides. Using pharmacological inhibitors, we show that elastin peptide treatment leads to activation of the anti-apoptotic protein Akt that phosphorylates the pro-apoptotic protein Bad, the Foxo3a transcription factor and caspase-9. Importantly, the anti-apoptotic effects of elastin peptides were persistent in time. Our results suggest that elastin peptides could be potent cell survival factors. PMID- 19559026 TI - Imaging membrane intercalating near infrared dyes to track multiple cell populations. AB - Given the increasing interest in understanding in vivo migration of different cell types, it would be useful to have a simple method for tracking multiple cell populations in animals. Here we evaluated near infrared (NIR) dyes that intercalate into cell membranes as cell tracking labels, using both high throughput and high-resolution methods. We tracked cells in tissues containing significant autofluorescence. CellVue Burgundy (ex 683/em 707) and CellVue NIR815 (ex 786/em 814) are especially useful because their spectral properties match the laser and detectors of the LI-COR laser scanner. After labeling cells ex vivo and injecting them into tumor-bearing mice, the distribution of cells in tumor and organs could be quantified in tissue sections with high throughput by scanning many slides at once. For example, we compared brain tumor infiltration and organ distribution of naive and activated lymphocytes in single animals. High resolution microscopic examination of the same tissues could be done by a relatively inexpensive modification of an epifluorescence microscope using a custom designed diode laser light source. Light emitting diodes that emit 685 nm and 780 nm light allowed microscopic visualization of the NIR labeled cells in tissues. The NIR dye-labeled cells were visualized with a greater signal/noise ratio compared to visible wavelength dyes such as CFSE, because of the low levels of autofluorescence in the NIR range. We also describe a simple modification of immunohistochemical procedures that allows combined visualization of the hydrophobic NIR dyes and antibody probes of cell markers in unfixed tissue. In combination these techniques will facilitate cell tracking in vivo. PMID- 19559027 TI - The structure of bacteriophage phiCb5 reveals a role of the RNA genome and metal ions in particle stability and assembly. AB - The structure of the Leviviridae bacteriophage phiCb5 virus-like particle has been determined at 2.9 A resolution and the structure of the native bacteriophage phiCb5 at 3.6 A. The structures of the coat protein shell appear to be identical, while differences are found in the organization of the density corresponding to the RNA. The capsid is built of coat protein dimers and in shape corresponds to a truncated icosahedron with T = 3 quasi-symmetry. The capsid is stabilized by four calcium ions per icosahedral asymmetric unit. One is located at the symmetry axis relating the quasi-3-fold related subunits and is part of an elaborate network of hydrogen bonds stabilizing the interface. The remaining calcium ions stabilize the contacts within the coat protein dimer. The stability of the phiCb5 particles decreases when calcium ions are chelated with EDTA. In contrast to other leviviruses, phiCb5 particles are destabilized in solution with elevated salt concentration. The model of the phiCb5 capsid provides an explanation of the salt induced destabilization of phiCb5, since hydrogen bonds, salt bridges and calcium ions have important roles in the intersubunit interactions. Electron density of three putative RNA nucleotides per icosahedral asymmetric unit has been observed in the phiCb5 structure. The nucleotides mediate contacts between the two subunits forming a dimer and a third subunit in another dimer. We suggest a model for phiCb5 capsid assembly in which addition of coat protein dimers to the forming capsid is facilitated by interaction with the RNA genome. The phiCb5 structure is the first example in the levivirus family that provides insight into the mechanism by which the genome-coat protein interaction may accelerate the capsid assembly and increase capsid stability. PMID- 19559028 TI - Relation of intracellular signal levels and promoter activities in the gal regulon of Escherichia coli. AB - Transcription of many genes is regulated by combinations of multiple signals. In Escherichia coli, combinatorial control is typical in the case of operons related to utilization of different sugars in the absence of glucose. To understand regulation of the transport and metabolic pathways in the galactose system, we measured activities of the six gal regulon promoters simultaneously, using an in vitro transcription system containing purified components. Input functions were computed on the basis of the experimental measurements. We observed four different shapes of input functions. From the results, we can conclude that the structure of the regulatory network is insufficient for the determination of signal integration. It is the actual structure of the promoter and regulatory region, the mechanism of transcription regulation, and the interplay between transcription factors that shape the input function to be suitable for adaptation. PMID- 19559030 TI - Crystal structure and putative mechanism of 3-methylitaconate-delta-isomerase from Eubacterium barkeri. AB - 3-Methylitaconate-Delta-isomerase (Mii) participates in the nicotinate fermentation pathway of the anaerobic soil bacterium Eubacterium barkeri (order Clostridiales) by catalyzing the reversible conversion of (R)-3-methylitaconate (2-methylene-3-methylsuccinate) to 2,3-dimethylmaleate. The enzyme is also able to catalyze the isomerization of itaconate (methylenesuccinate) to citraconate (methylmaleate) with ca 10-fold higher K(m) but > 1000-fold lower k(cat). The gene mii from E. barkeri was cloned and expressed in Escherichia coli. The protein produced with a C-terminal Strep-tag exhibited the same specific activity as the wild-type enzyme. The crystal structure of Mii from E. barkeri has been solved at a resolution of 2.70 A. The asymmetric unit of the P2(1)2(1)2(1) unit cell with parameters a = 53.1 A, b = 142.3 A, and c = 228.4 A contains four molecules of Mii. The enzyme belongs to a group of isomerases with a common structural feature, the so-called diaminopimelate epimerase fold. The monomer of 380 amino acid residues has two topologically similar domains exhibiting an alpha/beta-fold. The active site is situated in a cleft between these domains. The four Mii molecules are arranged as a tetramer with 222 symmetry for the N terminal domains. The C-terminal domains have different relative positions with respect to the N-terminal domains resulting in a closed conformation for molecule A and two distinct open conformations for molecules B and D. The C-terminal domain of molecule C is disordered. The Mii active site contains the putative catalytic residues Lys62 and Cys96, for which mechanistic roles are proposed based on a docking experiment of the Mii substrate complex. The active sites of Mii and the closely related PrpF, most likely a methylaconitate Delta-isomerase, have been compared. The overall architecture including the active-site Lys62, Cys96, His300, and Ser17 (Mii numbering) is similar. This positioning of (R)-3 methylitaconate allows Cys96 (as thiolate) to deprotonate C-3 and (as thiol) to donate a proton to the methylene carbon atom of the resulting allylic carbanion. Interestingly, the active site of isopentenyl diphosphate isomerase type I also contains a cysteine that cooperates with glutamate rather than lysine. It has been proposed that the initial step in this enzyme is a protonation generating a tertiary carbocation intermediate. PMID- 19559031 TI - Structural properties of neurofilament sidearms: sequence-based modeling of neurofilament architecture. AB - Neurofilaments (NFs) are essential cytoskeletal filaments that impart mechanical integrity to nerve cells. They are assembled from three distinct molecular mass proteins that bind to each other to form a 10-nm-diameter filamentous rod with sidearm extensions. The sidearms are considered to play a critical role in modulating interfilament spacing and axonal caliber. However, the precise mechanism by which NF protrusions regulate axonal diameter remains to be well understood. In particular, the role played by individual NF protrusions in specifying interfilament distances is yet to be established. To gain insight into the role of individual proteins, we investigated the structural organization of NF architecture under different phosphorylation conditions. To this end, a physically motivated sequence-based coarse-grain model of NF brush has been developed based on the three-dimensional architecture of NFs. The model incorporates the charge distribution of sidearms, including charges from the phosphorylation sites corresponding to Lys-Ser-Pro repeat motifs. The model also incorporates the proper grafting of the real NF sidearms based on the stoichiometry of the three subunits. The equilibrium structure of the NF brush is then investigated under different phosphorylation conditions. The phosphorylation of NF modifies the structural organization of sidearms. Upon phosphorylation, a dramatic change involving a transformation from a compact conformation to an extended conformation is found in the heavy NF (NF-H) protein. However, in spite of extensive phosphorylation sites present in the NF-H subunit, the tails of the medium NF subunit are found to be more extended than the NF-H sidearms. This supports the notion that medium NF protrusions are critical in regulating NF spacings and, hence, axonal caliber. PMID- 19559029 TI - A 5'UTR-spliced mRNA isoform is specialized for enhanced HIV-2 gag translation. AB - Full-length unspliced genomic RNA plays critical roles in HIV replication, serving both as mRNA for the synthesis of the key viral polyproteins Gag and Gag Pol and as genomic RNA for encapsidation into assembling viral particles. We show that a second gag mRNA species that differs from the genomic RNA molecule by the absence of an intron in the 5' untranslated region (5'UTR) is produced during HIV 2 replication in cell culture and in infected patients. We developed a cotransfection system in which epitopically tagged Gag proteins can be traced back to their mRNA origins in the translation pool. We show that a disproportionate amount of Gag is translated from 5'UTR intron-spliced mRNAs, demonstrating a role for the 5'UTR intron in the regulation of gag translation. To further characterize the effects of the HIV-2 5'UTR on translation, we fused wild-type, spliced, or mutant leader RNA constructs to a luciferase reporter gene and assayed their translation in reticulocyte lysates. These assays confirmed that leaders lacking the 5'UTR intron increased translational efficiency compared to that of the unspliced leader. In addition, we found that removal or mutagenesis of the C-box, a pyrimidine-rich sequence located in the 5'UTR intron and previously shown to affect RNA dimerization, also strongly influenced translational efficiency. These results suggest that the splicing of both the 5'UTR intron and the C-box element have key roles in regulation of HIV-2 gag translation in vitro and in vivo. PMID- 19559033 TI - Nerve growth factor-mediated paracrine regulation of hepatic stellate cells by multipotent mesenchymal stromal cells. AB - AIMS: Multipotent mesenchymal stromal cells (MSC) have been reported to prevent the development of liver fibrosis and have emerged as a promising strategy for cell-based therapy. However, the underlying therapeutic mechanism remains unclear. Hepatic stellate cells (SC) activation is a pivotal event in the development of liver fibrosis. MAIN METHODS: We hypothesized that MSC play an important role in regulating SC proliferation and apoptosis through paracrine mechanisms. To investigate the paracrine interactions between MSC and SC, a co culture experimental model was developed using human MSC (hMSC) and human SC (hSC). KEY FINDINGS: We demonstrate that hMSC and hSC both express nerve growth factor (NGF) receptor p75. Results acquired from transwell co-culture experiments using hSC and hMSC showed that hMSC secrete NGF, which enhances hSC apoptosis. Transcription factor nuclear factor kappa B (NF-KappaB) and B cell leukemia-xl (Bcl-xl) take part in the process. SIGNIFICANCE: These findings demonstrated that hMSC indirectly modulate activated hSC in vitro via NGF-mediated signaling cascades and provide a potential mechanism of how transplanted MSC are effective in treating liver fibrosis. PMID- 19559032 TI - High-yield heterologous expression of wild type and mutant Ca(2+) ATPase: Characterization of Ca(2+) binding sites by charge transfer. AB - High-yield heterologous SERCA1 (Ca(2+) ATPase) expression was obtained in COS-1 cells infected with recombinant adenovirus vector (rAdSERCA). Higher transcription and expression were obtained in the presence of a His(6) tag at the amino terminus, as compared with a His(6) tag at the carboxyl SERCA terminus, or no tag. The expressed protein was targeted extensively to intracellular membranes. Optimal yield of functional Ca(2+) ATPase corresponded to 10% of total protein, with phosphoenzyme levels, catalytic turnover and Ca(2+) transport identical with those of native SERCA1. This recombinant membrane-bound (detergent free) enzyme was used for characterization of Ca(2+) binding at the two specific transmembrane sites (ATP-free) by measurements of net charge transfer upon Ca(2+) binding to the protein, yielding cooperative isotherms (K(1)=5.9+/-0.5x10(5) M( 1) and K(2)=5.7+/-0.3x10(6) M(-1)). Non-cooperative binding of only one Ca(2+), and loss of ATPase activation, were observed following E309 mutation at site II. On the other hand, as a consequence of the site II mutation, the affinity of site I for Ca(2+) was increased (K=4.4+/-0.2x10(6) M(-1)). This change was due to a pK(a) shift of site I acidic residues, and to contributions of oxygen functions from empty site II to Ca(2+) binding at site I. No charge movement was observed following E771Q mutation at site I, indicating no Ca(2+) binding to either site. Therefore, calcium occupancy of site I is required to trigger cooperative binding to site II and catalytic activation. In the presence of millimolar Mg(2+), the charge movement upon addition of Ca(2+) to WT ATPase was reduced by 50%, while it was reduced by 90% when Ca(2+) was added to the E309Q/A mutants, demonstrating that competitive Mg(2+) binding can occur at site I but not at site II. PMID- 19559034 TI - Targeting pain-depressed behaviors in preclinical assays of pain and analgesia: drug effects on acetic acid-depressed locomotor activity in ICR mice. AB - AIMS: Pain depresses expression of many behaviors, and one goal of analgesic treatment is to restore pain-depressed behaviors. Assays that focus on pain depressed behaviors may contribute to preclinical assessment of candidate analgesics. MAIN METHODS: This study compared effects of the mu opioid receptor agonist morphine (an acknowledged analgesic), the dopamine receptor antagonist haloperidol (a non-analgesic sedative), the adenosine receptor antagonist caffeine (a non-analgesic stimulant) and the neurokinin-1 receptor antagonist CJ 11,974-01 (a candidate analgesic) on acetic acid-induced writhing (a traditional pain-stimulated behavior) and acetic acid-induced suppression of locomotor activity (a pain-depressed behavior) in male ICR mice. Drug effects on non depressed (baseline) locomotor activity were also examined. KEY FINDINGS: I.P. administration of acetic acid (0.18-1%) was equipotent in stimulating writhing and depressing locomotor activity. Morphine blocked both acid-induced stimulation of writhing and depression of locomotion, although it was 56-fold less potent in the assay of acid-depressed locomotion. Haloperidol and CJ 11,974-01 decreased acid-stimulated writhing, but failed to block acid-induced depression of locomotion. Caffeine had no effect on acid-stimulated writhing or acid-depressed locomotor activity, although it did increase non-depressed locomotion. Thus, morphine was the only drug to block both acid-stimulated writhing and acid depressed locomotion. SIGNIFICANCE: Complementary assays of pain-stimulated and pain-depressed behaviors may improve the predictive validity of preclinical studies that assess candidate analgesic drugs. The low potency of morphine to block acid-induced depression of locomotion suggests that locomotor activity may be a relatively insensitive measure for studies of pain-depressed behavior. PMID- 19559036 TI - Identification of a novel transmembrane domain involved in the negative modulation of mGluR1 using a newly discovered allosteric mGluR1 antagonist, 3 cyclohexyl-5-fluoro-6-methyl-7-(2-morpholin-4-ylethoxy)-4H-chromen-4-one. AB - Currently tested allosteric modulators for metabotropic glutamate receptor 1 (mGluR1) are known to regulate the activity of mGluR1 mainly through transmembrane (TM) domain 6 and/or 7. We identified a novel interaction site, N760 in TM5, which negatively regulates activation of mGluR1 with a newly discovered selective mGluR1 antagonist, 3-cyclohexyl-5-fluoro-6-methyl-7-(2 morpholin-4-ylethoxy)-4H-chromen-4-one (CFMMC). CFMMC inhibited L-glutamate induced intracellular Ca(2+) mobilization ([Ca(2+)]i) in Chinese hamster ovary (CHO) cells expressing recombinant human mGluR1a with IC(50) value of 50 nM, whereas it did not inhibit [Ca(2+)]i in CHO cells expressing human mGluR5a (IC(50); >10 microM). To identify the amino acid residues critical for antagonism of CFMMC, we constructed various point mutants of human mGluR1 and evaluated them in [Ca(2+)]i assays. The inhibitory effects of CFMMC were significantly affected in point mutations of either I725 in TM4 or N760 in TM5, as well as mutations of W798, F801 and Y805 in TM6 or T815 in TM7. Further studies revealed that antagonistic activities of not only CFMMC but also other, structurally unrelated, mGluR1 antagonists such as 6-amino-N-cyclohexyl-N,3-dimethylthiazolo[3,2 a]benzimidazole-2-carboxamide (YM-298198) and Compound 1 were reduced in N760 mutated mGluR1a. These results indicate that some mGluR1 allosteric antagonists require N760 in TM5 to demonstrate negative modulation of mGluR1 in addition to the reported amino acid residues in TM6 and TM7. PMID- 19559035 TI - Age-related LH surge dysfunction correlates with reduced responsiveness of hypothalamic anteroventral periventricular nucleus kisspeptin neurons to estradiol positive feedback in middle-aged rats. AB - Female reproductive aging in rats is characterized by reduced gonadotropin releasing hormone (GnRH) neuronal activation under estradiol positive feedback conditions and a delayed and attenuated luteinizing hormone (LH) surge. The newly identified excitatory neuropeptide kisspeptin is proposed to be a critical mediator of the pubertal transition and the ovarian steroid-induced LH surge. We previously showed that estradiol induces less kisspeptin mRNA expression in the anterior hypothalamus [anatomical location of anteroventral periventricular nucleus (AVPV)] in middle-aged than in young rats and intrahypothalamic infusion of kisspeptin restores LH surge amplitude in middle-aged females. Thus, reduced kisspeptin neurotransmission may contribute to age-related LH surge abnormalities. This study tested the hypothesis that middle-aged females will also exhibit reduced numbers of kisspeptin immunopositive neurons in the AVPV under estradiol positive feedback conditions. Using immunohistochemistry, we demonstrate that middle-aged females primed with ovarian steroids have fewer AVPV kisspeptin immunopositive neurons than young females. Age did not affect kisspeptin mRNA expression in the pituitary, numbers of kisspeptin immunopositive neurons in the arcuate nucleus, or estradiol-dependent reductions in kisspeptin mRNA expression in the posterior hypothalamus (containing the arcuate nucleus). These data strongly suggest that age-related LH surge dysfunction results, in part, from a reduced sensitivity of AVPV kisspeptin neurons to estradiol and hence decreased availability of AVPV kisspeptin neurons to activate GnRH neurons under positive feedback conditions. PMID- 19559038 TI - A method for acetonitrile-free microcystin analysis and purification by high performance liquid chromatography, using methanol as mobile phase. AB - Cyanobacteria produce a wide range of potent toxins, including hepatotoxic microcystins. HPLC methods for microcystin analysis and purification almost invariably include acetonitrile in the elution gradient mobile phase. The recent, acute, global acetonitrile shortage requires that adequate methods are available for microcystin analysis and purification without the need for acetonitrile. Here we present a convenient methanol-based method for effective HPLC analysis and purification of the toxins, with full separation of a range of microcystin variants. PMID- 19559037 TI - N-acetylaspartylglutamate (NAAG) inhibits intravenous cocaine self-administration and cocaine-enhanced brain-stimulation reward in rats. AB - Pharmacological activation of group II metabotropic glutamate (mGlu2 and mGlu3) receptors inhibits reward-seeking behavior and/or rewarding efficacy induced by drugs (cocaine, nicotine) or natural rewards (food, sucrose). In the present study, we investigated whether elevation of brain N-acetylaspartylglutamate (NAAG), an endogenous group II mGlu receptor agonist, by the NAAG peptidase inhibitor 2-PMPA attenuates cocaine's rewarding effects, as assessed by intravenous cocaine self-administration and intracranial electrical brain stimulation reward (BSR) in rats. Systemic administration of 2-PMPA (10, 30, 100 mg/kg, i.p.) or intranasal administration of NAAG (100, 300 microg/10 microl/nostril) significantly inhibited intravenous cocaine self-administration under progressive-ratio (PR), but not under fixed-ratio 2 (FR2), reinforcement conditions. In addition, 2-PMPA (1, 10, 30 mg/kg, i.p) or NAAG (50, 100 microg/10 microl/nostril) significantly inhibited cocaine-enhanced BSR, but not basal BSR. Pretreatment with LY341495 (1 mg/kg, i.p.), a selective mGlu2/3 receptor antagonist, prevented the inhibitory effects produced by 2-PMPA or NAAG in both the self-administration and BSR paradigms. In vivo microdialysis demonstrated that 2-PMPA (10, 30, 100 mg/kg) dose-dependently attenuated cocaine-enhanced extracellular dopamine (DA) in the nucleus accumbens (NAc). 2-PMPA alone inhibited basal NAc DA release, an effect that was prevented by LY341495. These findings suggest that systemic administration of 2-PMPA or intranasal administration of NAAG inhibits cocaine's rewarding efficacy and cocaine-enhanced NAc DA - likely by activation of presynaptic mGlu2/3 receptors in the NAc. These data suggest a potential utility for 2-PMPA or NAAG in the treatment of cocaine addiction. PMID- 19559039 TI - Effect of calcium on the hemolytic activity of Stichodactyla helianthus toxin sticholysin II on human erythrocytes. AB - Sticholysin II (St II) is a toxin from the sea anemona Stichodactyla helianthus that produces erythrocytes lysis at low concentration and its activity depends on the presence of calcium. Calcium may act modifying toxin interaction with erythrocyte membranes or activating cellular processes which may result in a modified St II lytic action. In this study we are reporting that, in the presence of external K(+), extracellular calcium decreased St II activity on erythrocytes. On the other hand an increase of intracellular calcium promotes Sty II lytic activity. The effect of intracellular calcium was specifically studied in relation to membrane lipid translocation elicited by scramblases and how this action influence St II lytic activity on erythrocytes. We used 0.5 mmol/L calcium and 10 mmol/L A23187, as calcium ionophore, for scramblases activation and found increased St II activity associated to increase of intracellular calcium. N-ethyl maleimide (activator) and 4,4'-diisothiocyanatostilbene-2,2'-disulfonate (inhibitor) were used as scramblases modulators in the assays which produced an increase and a decrease of the calcium effect, respectively. Results reported suggest an improved St II membrane pore-forming capacity promoted by intracellular calcium associated to membrane phospholipids translocation. PMID- 19559040 TI - Limited uptake of the cyanobacterial toxin cylindrospermopsin by Vero cells. AB - Cylindrospermopsin (CYN) is a cyanobacterial toxin increasingly found in drinking water sources worldwide. Toxicity studies have shown CYN can induce effects in a range of different cell types with primary hepatocytes consistently shown to be the most sensitive cellular model. How CYN enters the intracellular environment is not clear, although the size and hydrophilic nature of the toxin suggest it would not readily cross a lipid bilayer. In this study, a Vero cell line expressing green fluorescent protein (GFP) was used to monitor for CYN uptake based on the toxin's potent effects on protein synthesis. Effects on the GFP signal were compared with inhibitors cycloheximide (CHEX) and emetine. While CYN potency was demonstrated in a cell-free system (CYN>CHEX>emetine) it was considerably reduced in the Vero-GFP cell model (CHEX, emetine>>CYN). In contrast to other inhibitors, CYN effects on GFP signal increased 6 fold over 4-24 h incubation indicating slow, progressive uptake of the toxin. Confirming that the uptake process is not energy dependent CYN entry also occurred at 4 degrees C, while competition experiments excluded the uracil nucleobase transporter system as potential mechanism for CYN uptake. Dilution of media enhanced CYN uptake by Vero-GFP cells although mechanism by which this occurred is unknown. PMID- 19559041 TI - Brevetoxin B is a clastogen in rats, but lacks mutagenic potential in the SP 98/100 Ames test. AB - Brevetoxins are polyether toxins produced by the dinoflagellate Karenia brevis that are released into the air and are known to cause respiratory hemorrhage in manatees and irritation in humans. Brevetoxin has been previously reported to cause DNA breakage and chromosomal aberrations in in vitro cell assays. The toxin is subject to epoxidation reaction and the formation of nucleic acid adducts in cultured lung fibroblasts and in lung tissue after intratracheal administration to rats. We have exposed rats intratracheally to brevetoxin B (45 microg/kg) and analyzed liver cells for DNA fragmentation using a comet assay. Brevetoxin B (PbTx2) treated rats showed a two to three-fold increase in the amount of DNA in the comet tails, indicating that brevetoxin has in vivo clastogenic activity. We next tested brevetoxin B for mutagenic activity using the Ames 98/100 mutagenesis assay. Brevetoxin B at concentrations from 0.064 to 200 microg/mL failed to cause histidine revertants. Oxidative metabolism of brevetoxin B resulting from Aroclor 1259-induced rat liver microsomes also failed to cause histidine revertants. Finally, direct application of the brevetoxin B epoxide (PbTx6) in the Ames 98/100 assay at concentrations from 0.064 to 200 microg/mL failed to induce histidine revertants. These studies indicate that brevetoxin B retains clastogenic activity after intratracheal administration to the rat. Although brevetoxin B has been shown to form nucleic acid adducts in the lung, neither brevetoxin B nor its epoxide metabolite has mutagenic potential as assessed by the Ames 98/100 test. PMID- 19559042 TI - Protective effect of a Potentilla anserine polysaccharide on oxidative damages in mice. AB - Potentilla anserine polysaccharide (PAP) was studied in vivo to investigate its antioxidant activity using the model of dexamethasone-induced oxidative stress in mice. The investigation demonstrated that PAP at 50, 100 or 200mg/kg body weight for 7 days respectively increased thymus index and spleen index, glutathione level, superoxidase dismutase activity and total antioxidant capacity in both thymus and spleen and decreased the content of H(2)O(2) in spleen and NO in both thymus and spleen of mice. The results revealed that PAP was able to overcome dexamethasone-induced oxidative stress and might play an important role in repairs of oxidative damage in immunological system. PMID- 19559043 TI - Autism: a world changing too fast for a mis-wired brain? AB - Disorders in verbal and emotional communication and imitation, social reciprocity and higher order cognition observed in individuals with autism spectrum disorders (ASD) are presented here as phenotypic expressions of temporo-spatial processing disorders (TSPDs). TSPDs include various degrees of disability in (i) processing multi-sensory dynamic stimuli online, (ii) associating them into meaningful and coherent patterns and (iii) producing real-time sensory-motor adjustments and motor outputs. In line with this theory, we found that slowing down the speed of facial and vocal events enhanced imitative, verbal and cognitive abilities in some ASD children, particularly those with low functioning autism. We then argue that TSPDs may result from Multi-system Brain Disconnectivity-Dissynchrony (MBD), defined as an increase or decrease in functional connectivity and neuronal synchronization within/between multiple neurofunctional territories and pathways. Recent functional magnetic resonance imaging (fMRI) and electrophysiological studies supporting MBD are outlined. Finally, we review the suspected underlying neurobiological mechanisms of MBD as evidenced in neuroimaging, genetic, environmental and epigenetic studies. Overall, our TSPD/MBD approach to ASD may open new promising avenues for a better understanding of neuro-physio psychopathology of ASD and clinical rehabilitation of people affected by these syndromes. PMID- 19559044 TI - Motivation on the Mediterranean: reward, compulsions and habit formation. AB - The 2007 Motivational Neuronal Networks meeting, held in Porquerolles, France was organized to generate debate and discussion on issues relating to reward, compulsion, and habit formation. The conference consisted primarily of four workshops that brought researchers from a wide variety of fields together in an informal atmosphere designed to facilitate interaction. This report is based on the detailed notes taken during the wide-ranging discussions, and summarizes major areas of both consensus and disagreement, as well research topics that are likely to be high priorities in the years to come. PMID- 19559046 TI - The use of proton magnetic resonance spectroscopy in PTSD research--meta-analyses of findings and methodological review. AB - Different neuroimaging techniques provided evidence for structural and functional brain alterations in posttraumatic stress disorder (PTSD). Due to technical improvements, especially concerning localization techniques and more reliable analysis methods, one technique, proton magnetic resonance spectroscopy ((1)H MRS), has increasingly become of interest because it allows further insight into metabolic mechanisms that may contribute to these alterations. The aim of this article is, therefore, to review recent studies utilizing (1)H-MRS of the hippocampus and other brain structures in PTSD. Using meta-analytic methods, we attempted to answer the question if PTSD, as compared to different types of control samples, is accompanied by altered neurometabolite ratios and concentrations in the tissue of different brain regions. A second intent was to review methodological aspects to advise on a minimal standard for reliable results with respect to the application of (1)H-MRS in PTSD. Finally, we discussed the implications of the findings with respect to current PTSD models and future research. PMID- 19559045 TI - Dopamine and psychosis: theory, pathomechanisms and intermediate phenotypes. AB - Schizophrenia is a chronic, severe, and disabling brain disorder arising from the adverse interaction of predisposing risk genes and environmental factors. The psychopathology is characterized by a wide array of disturbing cognitive, emotional, and behavioral symptoms that interfere with the individual's capacity to function in society. Contemporary pathophysiological models assume that psychotic symptoms are triggered by a dysregulation of dopaminergic activity in the brain, a theory that is tightly linked to the serendipitous discovery of the first effective antipsychotic agents in the early 1950s. In recent years, the availability of modern neuroimaging techniques has significantly expanded our understanding of the key mediator circuits that bridge the gap between genetic susceptibility and clinical phenotype. This paper discusses the pathophysiological concepts, molecular mechanisms and neuroimaging evidence that link psychosis to disturbances in dopamine neurotransmission. PMID- 19559047 TI - Evaluation of side effects through selective ablation of the mu opioid receptor expressing descending nociceptive facilitatory neurons in the rostral ventromedial medulla with dermorphin-saporin. AB - Descending facilitation from the rostral ventromedial medulla (RVM) contributes to some pathological pain states. The intra-RVM microinjection with dermorphin saporin could specifically abolish this facilitation in rodent models by selectively ablating the RVM neurons expressing mu opioid receptors. Thus, this targeted lesion may be an alternative mechanism-based approach for intractable pain. This research was performed to investigate potential side effects after a single intra-RVM application of dermorphin-saporin in rats. Results showed though some acute cardiovascular signs were observed with dermorphin-saporin, the treatment exhibited no long-lasting significant influence on some physiological functions for up to 3-month observation period, including normal sensory function, locomotor activity, ingestive behaviors, body weight, rectal temperature, respiratory rate, heart rate, systolic blood pressure, cardiac structure and function. Moreover, there were only mild microglial responses on day 7 post-microinjection, while no significant increase in the immunostaining of astrocytes and no noticeable up-regulation in the production of proinflammatory cytokines were detected in the RVM treated with dermorphin-saporin. Taken together, these data would suggest that this selective ablation of mu opioid receptor bearing descending facilitatory neurons in the RVM with dermorphin saporin did not elicit the long-standing evident adverse toxicity in terms of some physiological parameters and neurochemical alterations we determined, plausibly providing us a safe and reliable approach to treat some intractable pain. PMID- 19559048 TI - Retrograde labeling of phrenic motoneurons by intrapleural injection. AB - Studies of motoneuron plasticity during development or in response to injury or disease rely on the ability to correctly identify motoneurons innervating specific muscle groups. Commonly, injections of retrograde tracer molecules into a target muscle or into a transected nerve are used to label specific motoneuron pools. However, intramuscular injection does not consistently label all motoneurons in the target pool; and either injection site may involve additional surgical procedures and muscle or nerve perturbations. For instance, retrograde labeling of phrenic motoneurons by injection into the diaphragm muscle is commonly employed in studies of plasticity in respiratory motor control. Diaphragm intramuscular injection involves a laparotomy, and this additional surgery may limit the ability to conduct labeling studies particularly in small animals. In the present study, we provide validation of a novel method for phrenic motoneuron labeling using intrapleural injection of Alexa 488-conjugated cholera toxin subunit B. Only phrenic motoneurons were labeled in the cervical spinal cord as verified by co-staining with rhodamine-conjugated dextran injected into the diaphragm muscle or applied via phrenic nerve dip. Thoracic intercostal motoneurons and some dorsal root ganglia neurons were also labeled by intrapleural injection, but there was no evidence of trans-synaptic labeling. Phrenic motoneuron labeling was not present if the phrenic nerve was transected prior to intrapleural injection. This novel method is ideally suited for morphological studies and analyses of mRNA expression in isolated phrenic motoneurons using techniques such as laser capture microdissection. PMID- 19559049 TI - Simple and cost-effective hardware and software for functional brain mapping using intrinsic optical signal imaging. AB - We describe a simple and low-cost system for intrinsic optical signal (IOS) imaging using stable LED light sources, basic microscopes, and commonly available CCD cameras. IOS imaging measures activity-dependent changes in the light reflectance of brain tissue, and can be performed with a minimum of specialized equipment. Our system uses LED ring lights that can be mounted on standard microscope objectives or video lenses to provide a homogeneous and stable light source, with less than 0.003% fluctuation across images averaged from 40 trials. We describe the equipment and surgical techniques necessary for both acute and chronic mouse preparations, and provide software that can create maps of sensory representations from images captured by inexpensive 8-bit cameras or by 12-bit cameras. The IOS imaging system can be adapted to commercial upright microscopes or custom macroscopes, eliminating the need for dedicated equipment or complex optical paths. This method can be combined with parallel high resolution imaging techniques such as two-photon microscopy. PMID- 19559050 TI - An improved method for protecting and fixing the lumbar catheters placed in the spinal subarachnoid space of rats. AB - The chronic intrathecal catheterization of the subarachnoid space in the rat is a very useful model in the research of neuroscience, anesthesiology, neurotoxicity, and etc. To avoid high postoperative mortality and neurological impairment, the catheterization from the lumbar interspace has been widely used in recent years. But one of the most discouraging disadvantages is that the external portion of the chronic indwelling catheters is easily damaged by rats. Besides, the short length of catheter left in the subarachnoid space in addition to its slimness makes it difficult for proper fixation. To improve the abovementioned situations, we designed a protector to protect the external portion of the catheters and developed a simple method for better fixation. Results suggest that this is an excellent method for protecting and fixing the lumbar catheters placed in the subarachnoid space of rats, and this method did not affect the sensory and motor function of the rat. The general conditions of the rats were unswayed. PMID- 19559051 TI - MRI-guided stereotaxic targeting in pigs based on a stereotaxic localizer box fitted with an isocentric frame and use of SurgiPlan computer-planning software. AB - We present a stereotaxic procedure enabling MRI-guided isocentric stereotaxy in pigs. The procedure is based on the Leksell stereotaxic arch principle, and a stereotaxic localizer box with an incorporated fiducial marking system (sideplates) defining a stereotaxic space similar to the clinical Leksell system. The obtained MRIs can be imported for 3D-reconstruction and coordinate calculation in the clinical stereotaxic software planning system (Leksell SurgiPlan, Elekta AB, Sweden). After MRI the sideplates are replaced by a modified Leksell arch accommodating clinical standard manipulators for isocentric placement of DBS-electrodes, neural tracers and therapeutics in the calculated target coordinates. The mechanical accuracy of the device was within 0.3-0.5 mm. Stereotaxic MRIs were imported to the stereotaxic software planning system with a mean error of 0.4-0.5 mm and a max error of 0.8-0.9 mm. Application accuracy measured on a phantom and on inserted skull markers in nine pigs was within 1 mm in all planes. The intracerebral application accuracy found after placement of 10 manganese trajectories within the full extent of the intracerebral stereotaxic space in two minipigs was equally randomly distributed and within 0.7+/-0.4; 0.5+/-0.4; and 0.7+/-0.3mm in the X, Y, and Z plane. Injection of neural tracers in the subgenual gyrus of three minipigs and placement of encapsulated gene modified cells in four minipigs confirmed the accuracy and functionality of the described procedure. We conclude that the devised technique and instrumentation enable high-precision stereotaxic procedures in pigs that may benefit future large animal neuroscience research and outline the technical considerations for a similar stereotaxic methodology in other animals. PMID- 19559052 TI - Righting elicited by novel or familiar auditory or vestibular stimulation in the haloperidol-treated rat: rat posturography as a model to study anticipatory motor control. AB - External cues, including familiar music, can release Parkinson's disease patients from catalepsy but the neural basis of the effect is not well understood. In the present study, posturography, the study of posture and its allied reflexes, was used to develop an animal model that could be used to investigate the underlying neural mechanisms of this sound-induced behavioral activation. In the rat, akinetic catalepsy induced by a dopamine D2 receptor antagonist (haloperidol 5mg/kg) can model human catalepsy. Using this model, two experiments examined whether novel versus familiar sound stimuli could interrupt haloperidol-induced catalepsy in the rat. Rats were placed on a variably inclined grid and novel or familiar auditory cues (single key jingle or multiple key jingles) were presented. The dependent variable was movement by the rats to regain equilibrium as assessed with a movement notation score. The sound cues enhanced movements used to regain postural stability and familiar sound stimuli were more effective than unfamiliar sound stimuli. The results are discussed in relation to the idea that nonlemniscal and lemniscal auditory pathways differentially contribute to behavioral activation versus tonotopic processing of sound. PMID- 19559053 TI - Statistical significance of sequential firing patterns in multi-neuronal spike trains. AB - Sequential firings with fixed time delays are frequently observed in simultaneous recordings from multiple neurons. Such temporal patterns are potentially indicative of underlying microcircuits and it is important to know when a repeatedly occurring pattern is statistically significant. These sequences are typically identified through correlation counts. In this paper we present a method for assessing the significance of such correlations. We specify the null hypothesis in terms of a bound on the conditional probabilities that characterize the influence of one neuron on another. This method of testing significance is more general than the currently available methods since under our null hypothesis we do not assume that the spiking processes of different neurons are independent. The structure of our null hypothesis also allows us to rank order the detected patterns. We demonstrate our method on simulated spike trains. PMID- 19559054 TI - Assay development and high-throughput antiviral drug screening against Bluetongue virus. AB - Bluetongue virus (BTV) infection is one of the most important diseases of domestic livestock. There are no antivirals available against BTV disease. In this paper, we present the development, optimization and validation of an in vitro cell-based high-throughput screening (HTS) assay using the luminescent based CellTiter-Glo reagent to identify novel antivirals against BTV. Conditions of the cytopathic effect (CPE)-based assay were optimized at cell density of 5000 cells/well in medium containing 1% FBS and a multiplicity of infection at 0.01 in 384-well plate, with Z'-values > or = 0.70, Coefficient of Variations > or = 5.68 and signal-to-background ratio > or = 7.10. This assay was further validated using a 9532 compound library. The fully validated assay was then used to screen the 194,950 compound collection, which identified 693 compounds with >30% CPE inhibition. The 10-concentration dose response assay identified 185 structures with IC(50) < or =100 microM, out of which 42 compounds were grouped into six analog series corresponding to six scaffolds enriched within the active set compared to their distribution in the library. The CPE-based assay development demonstrated its robustness and reliability, and its application in the HTS campaign will make significant contribution to the antiviral drug discovery against BTV disease. PMID- 19559055 TI - A pharmacogenetic study of polymorphisms in interferon pathway genes and response to interferon-alpha treatment in chronic hepatitis B patients. AB - Certain host genetic polymorphisms in interferon (IFN) signaling pathway genes are reported to be associated with response to IFNalpha therapy. We studied 10 single nucleotide polymorphisms (SNPs) in IFN signaling pathway genes to examine their associations with response to IFN treatment in chronic hepatitis B (CHB) patients. Two hundred and forty-six IFNalpha treatment-naive CHB patients were enrolled for the present study; all received treatment with IFNalpha alone for 6 months, and the efficacy of the therapy was examined. Ten SNPs in 8 IFN signaling pathway genes were genotyped using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) protocol. There were no significant differences in allele frequencies and genotype distributions of the 10 SNPs between the response and non-response groups that underwent IFNalpha therapy. However, the frequency of a G-T-G-A 2',5'-oligoadenylate synthetase (OAS) haplotype was significantly higher in the non-response group than that in the response group (16.1% vs. 8.7%, p=0.015). Our study suggested that patients with a G-T-G-A OAS haplotype were less responsive to IFNalpha treatment. PMID- 19559057 TI - Ipsilateral responses of motor evoked potential correlated with the motor functional outcomes after cortical resection. AB - The aim of this study was to evaluate if ipsilateral motor evoked potential (MEP) elicited by transcranial magnetic stimulation (TMS) could provide neurosurgeons preoperatively with useful information regarding surgical procedure for patients with severe cerebral hemiatrophy or unilateral malformation. Thirteen epilepsy patients with severe cerebral hemiatrophy or unilateral malformation were studied before operation using MEPs recorded on bilateral abductor pollicis brevis (APBs) muscles, elicited by transcranial magnetic stimulation of the motor cortex. Ten subjects served as controls. RESULTS: (1) no ipsilateral MEP responses were recorded in all the 10 healthy subjects; (2) in the 13 patients, the results of MEPs could be divided into four types. Type A: in 3 patients bilateral MEPs were recorded when unaffected hemisphere was stimulated, while no responses were elicited when the affected hemisphere was stimulated. Type B: in another 3 patients, the MEPs were elicited from bilateral APB muscles when the unaffected hemisphere was stimulated, and the contralateral MEP was also elicited when the affected hemisphere was stimulated. Type C: in two patients contralateral MEP was elicited when the unaffected hemisphere was stimulated, while no MEP was induced in APB muscles of either side following the affected hemisphere stimulation. Type D: in the remaining 5 patients, contralateral magnetic MEPs were elicited either when the affected or the unaffected hemisphere was stimulated. Patients of type A, B and C received hemispherectomy showed no significant permanent motor functional deficit. Among the total 8 patients, 7 patients got seizure free after the operation. Patients of type D showed minor muscle strength decrease after localized cortical resection. Three out of 5 patients of type D got seizure free after the operation. Ipsilateral MEP response might be useful for neurosurgeons to plan appropriate surgical procedure which helps avoid post-operative motor deficits. PMID- 19559058 TI - Theophylline attenuates hippocampal blood flow responses induced by tooth pulp stimulation in rats. AB - In this study, we performed tests to determine whether tooth pulp stimulation (TPS) increases hippocampal blood flow (HBF), and if so, to investigate whether the increase in HBF is mediated via the activation of adenosine receptors. We measured HBF in urethane-anesthetized rats using laser Doppler flowmetry (LDF) and examined the effect of theophylline, a nonselective adenosine receptor antagonist, on TPS-induced HBF responses. TPS increased HBF, and its response was significantly attenuated by the intraperitoneal administration of theophylline (20 mg/kg). These results suggest that the HBF response induced by TPS may be, at least in part, produced through adenosine receptors. PMID- 19559059 TI - N-acetylcysteine alters apoptotic gene expression in axotomised primary sensory afferent subpopulations. AB - Novel approaches are required in peripheral nerve injury management because current surgical techniques, which do not address axotomy-induced neuronal death, lead to deficient sensory recovery. Sensory neuronal death has functional preference with cutaneous neurons dying in great numbers whilst muscle afferents survive axotomy. This offers the potential of comparing similar cell types that suffer distinct fates upon nerve injury. Here, a novel approach, combining in vivo rat nerve injury model with laser microdissection and quantitative real-time polymerase chain reaction, identifies crucial disparities in apoptotic gene expression attributable to subpopulations of differing sensory modalities and examines the response to N-acetylcysteine (NAC) therapy. We show that axotomised muscle afferent neurons survive injury due to a neuroprotective response which markedly downregulates Bax and caspase-3 mRNA. In contrast, axotomised cutaneous sensory neurons significantly upregulate caspase-3 and alter both Bcl-2 and Bax expression such that pro-apoptotic Bax predominates. N-Acetylcysteine (NAC) intervention promotes neuroprotection of cutaneous sensory neurons through considerable upregulation of Bcl-2 and downregulation of both Bax and caspase-3 mRNA. The data presented identifies differential activation of apoptotic genes in axotomised neuronal subpopulations. Furthermore, NAC therapy instigates apoptotic gene expression changes in axotomised neurons, thereby offering pharmacotherapeutic potential in the clinical treatment of nerve injury. PMID- 19559061 TI - Disruption of the OCH1 and MNN1 genes decrease N-glycosylation on glycoprotein expressed in Kluyveromyces lactis. AB - Glycoproteins secreted by the yeast Kluyveromyces lactis are usually modified by the addition at asparagines-linked glycosylation sites of heterogeneous mannan residues. The secreted glycoproteins in K. lactis that become hypermannosylated will bear a non-human glycosylation pattern and can adversely affect the half life, tissue distribution and immunogenicity of a therapeutic protein. Here, we describe engineering a K. lactis strain to produce non-hypermannosylated glycoprotein, decreasing the outer-chain mannose residues of N-linked oligosaccharides. We investigated and developed the method of two-step homologous recombination to knockout the OCH1 gene, encoding alpha1,6-mannosyltransferase and MNN1 gene, which is homologue of Saccharomyces cerevisiae MNN1, encoding a putative alpha1,3-mannosyltransferase. We found the Kloch1 mutant strain has a defect in hyperglycosylation, inability in adding mannose to the core oligosaccharide. The N-linked oligosaccharides assembled on a secretory glycoprotein, HSA/GM-CSF in Kloch1 mutant, contained oligosaccharide Man(13 14)GlcNAc(2), and in Kloch1 mnn1 mutant, contained oligosaccharide Man(9 11)GlcNAc(2), whereas those in the wild-type strain, consisted of oligosaccharides with heterogeneous sizes, Man(>30)GlcNAc(2). Taken together, these results indicated that KlOch1p plays a key role in the outer-chain mannosylation of N-linked oligosaccharides in K. lactis. The KlMnn1p, was proved to be certain contribution to the outer hypermannosylation, most possibly encodes alpha1,3-mannosyltransferase. Therefore, the Kloch1 and Kloch1 mnn1 mutants can be used as a foundational host to produce glycoproteins lacking the outer-chain hypermannoses and further maybe applicable to be a promising system for yeast therapeutic protein production. PMID- 19559062 TI - The rosettazyme: a synthetic cellulosome. AB - Cellulose is an attractive feedstock for biofuel production because of its abundance, but the cellulose polymer is extremely stable and its constituent sugars are difficult to access. In nature, extracellular multi-enzyme complexes known as cellulosomes are among the most effective ways to transform cellulose to useable sugars. Cellulosomes consist of a diversity of secreted cellulases and other plant cell-wall degrading enzymes bound to a protein scaffold. These scaffold proteins have cohesin modules that bind conserved dockerin modules on the enzymes. It is thought that the localization of these diverse enzymes on the scaffold allows them to function synergistically. In order to understand and harness this synergy smaller, simplified cellulosomes have been constructed, expressed, and reconstituted using truncated cohesin-containing scaffolds. Here we show that an 18-subunit protein complex called a rosettasome can be genetically engineered to bind dockerin-containing enzymes and function like a cellulosome. Rosettasomes are thermostable, group II chaperonins from the hyperthermo-acidophilic archaeon Sulfolobus shibatae, which in the presence of ATP/Mg(2+) assemble into 18-subunit, double-ring structures. We fused a cohesin module from Clostridium thermocellum to a circular permutant of a rosettasome subunit, and we demonstrate that the cohesin-rosettasomes: (1) bind dockerin containing endo- and exo-gluconases, (2) the bound enzymes have increased cellulose-degrading activity compared to their activity free in solution, and (3) this increased activity depends on the number and ratio of the bound glucanases. We call these engineered multi-enzyme structures rosettazymes. PMID- 19559060 TI - Methamphetamine treatment causes delayed decrease in novelty-induced locomotor activity in mice. AB - Methamphetamine (METH) is a psychostimulant that causes damage to dopamine (DA) axons and to non-monoaminergic neurons in the brain. The aim of the present study was to investigate short- and long-term effects of neurotoxic METH treatment on novelty-induced locomotor activity in mice. Male BALB/c mice, 12-14 weeks old, were injected with saline or METH (i.p., 7.5 mg/kg x 4 times, every 2 h). Behavior and neurotoxic effects were assessed at 10 days, 3 and 5 months following drug treatment. METH administration caused marked decreases in DA levels in the mouse striatum and cortex at 10 days post-drug. However, METH did not induce any changes in novelty-induced locomotor activity. At 3 and 5 months after treatment METH-exposed mice showed significant recovery of DA levels in the striatum and cortex. In contrast, these animals demonstrated significant decreases in locomotor activity at 5 months in comparison to aged-matched control mice. Further assessment of METH toxicity using TUNEL staining showed that the drug induced increased cell death in the striatum and cortex at 3 days after administration. Taken together, these data suggest that delayed deficits in novelty-induced locomotor activity observed in METH-exposed animals are not due to neurodegeneration of DA terminals but to combined effects of METH and age dependent dysfunction of non-DA intrinsic striatal and/or corticostriatal neurons. PMID- 19559063 TI - Use of NaCl prevents aggregation of recombinant COMP-angiopoietin-1 in Chinese hamster ovary cells. AB - To investigate the effect of hyperosmotic medium on production and aggregation of the variant of Angiopoietin-1 (Ang1), cartilage oligomeric matrix protein (COMP) Ang1, in recombinant Chinese hamster ovary (CHO) cells, CHO cells were cultivated in shaking flasks. NaCl and/or sorbitol were used to raise medium osmolality in the range of 300-450mOsm/kg. The specific productivity of COMP-Ang1, q(COMP Ang1), increased as medium osmolality increased. At NaCl-450mOsm/kg, the q(COMP Ang1) was 7.7-fold higher than that at NaCl-300mOsm/kg, while, at sorbitol 450mOsm/kg, it was 2.9-fold higher than that at sorbitol-300mOsm/kg. This can be attributed to the increased relative mRNA level of COMP-Ang1 at NaCl-450mOsm/kg which was approximately 2.4-fold higher than that at sorbitol-450mOsm/kg. Western blot analysis showed that COMP-Ang1 aggregates started to occur in the late exponential phase of cell growth. When sorbitol was used to raise the medium osmolality, a severe aggregation of COMP-Ang1 was observed. On the other hand, when NaCl was used, the aggregation of COMP-Ang1 was drastically reduced at NaCl 400mOsm/kg. At NaCl-450mOsm/kg, the aggregation of COMP-Ang1 was hardly observed. This suggests that environmental conditions are critical for the aggregation of COMP-Ang1. Taken together, the use of NaCl-induced hyperosmotic medium to cell culture process turns out to be an efficient strategy for enhancing COMP-Ang1 production and reducing COMP-Ang1 aggregation. PMID- 19559064 TI - Cytotoxicity study of rock wool by cell magnetometric evaluation. AB - The cytotoxicity of rock wool (RW), an asbestos substitute, was evaluated by cell magnetometry. Alveolar macrophages were isolated from male Fisher rats. Following addition of triiron tetraoxide (Fe(3)O(4)) to macrophages, RW was added. Then, the remnant magnetic field strength was measured for 20min after magnetization by an external field. Relaxation, an indicator of decay of cytotoxicity, was observed by cell magnetometry immediately postmagnetization in the group to which RW was added. In general, materials phagocytosed by macrophages are ingested into phagosomes and digested while migrating. This migration of phagosomes occurs by polymerization and depolymerization of the cytoskeleton. As a result of evaluation, relaxation was not delayed by addition of RW, since RW caused no effect on the cytoskeleton. It was suggested that RW has no cytotoxicity as evaluated by cell magnetometry. PMID- 19559065 TI - Derivation of the minimal magnitude of the Critical Effect Size for continuous toxicological parameters from within-animal variation in control group data. AB - Assuming that temporal fluctuations in physiological parameters (e.g. haematology, biochemistry) in individual healthy non-exposed animals are non adverse, the minimal magnitude of the Critical Effect Size (CES) for a number of continuous parameters of toxicity studies was derived. A total of 36 studies (19 pharmaceutical preclinical studies in dogs and 17 chemical risk assessment studies in rats) were analysed to determine within-animal variation in their control groups. Minimal CES-values were derived for each group of studies, differentiating where necessary between strains and sexes, using the 2.5 percentile (lower limit) and/or 97.5 percentile (upper limit) of the distribution of the within-animal variation around the mean of each parameter. We concluded that minimal CES-values for continuous clinical chemistry and haematology parameters should be established separately per species, strain, sex and study duration investigated. Grouping of minimal CES-values, leading to more or less "general" values, seems possible for those parameters that are subject to tight homeostatic control and consequently show little within-animal variation. Nearly a quarter of the proposed CES-values is 5%, nearly a quarter range from 6% to 10%, a quarter is 15% or 20%, and nearly 30% of the proposed values is 20% of the mean of the control animals. PMID- 19559066 TI - Comments on serious anaphylaxis caused by nine Chinese herbal injections used to treat common colds and upper respiratory tract infections. AB - Reports describing severe allergic shock and fatality following treatment of a common cold or upper respiratory tract infection (URTI) with a Chinese herbal injection were collected. Our analysis of the risks associated with this treatment suggested that the potential risk of serious, or even lethal, anaphylaxis should preclude its use in treating common colds and URTIs. In light of our findings herein, we propose the following five suggestions for improving the clinical safety of delivering Chinese herbal injections as medical treatments. First, Chinese herbal injections should not be delivered in the clinic to treat patients in accordance with Bian zheng lun zhi (broad-spectrum application based on holistic Traditional Chinese Medicine (TCM) theory and methodology), but rather they should be administered to target specific indicated disease processes. Second, Chinese herbal injection indications should be based on the results of double-blind randomized controlled clinical trials. Third, Chinese herbal injections should be used only in cases involving severe disease or to rescue patients in critical condition; they should not be used to treat mild, relatively innocuous diseases, such as common colds and upper respiratory tract infections, given the risk of doing harm. Fourth, Chinese herbal injection formulas should include materials from only a single or a small number of plant sources in known quantities. Fifth, more studies examining the toxicology and allergenic potential of Chinese herbal injections are needed. PMID- 19559067 TI - A 13-week subchronic oral toxicity study of L-serine in rats. AB - A subchronic oral toxicity study was conducted to evaluate the safety of L-serine in Sprague-Dawley rats. The test article was administered once daily by gavage in male and female rats at dose levels of 0, 500, 1500, and 3000 mg/kg body weight/day for 13 weeks. Daily clinical signs, body weight, and food consumption were not affected by ingestion of the test article. There were no treatment related adverse effects on urinalysis, hematology, serum biochemistry, organ weights, gross and histopathological examination. It was concluded that the no observed-effect level (NOEL) for L-serine was 3000 mg/kg bw/day for both genders. PMID- 19559069 TI - Interactions of human ribosomal proteins S16 and S5 with an 18S rRNA fragment containing their binding sites. AB - Human ribosomal proteins S5e and S16e are the homologues of prokaryotic S7p and S9p, respectively. It was shown that S5e and S16e are capable of the specific binding with a rRNA transcript corresponding to the region of human 18S rRNA containing helices H28-30 and H41-43 (3Dm), which is homologous to the region in 16S rRNA containing the entire binding site for S7p and the major part of the site for S9p. We have studied binding of S5e and S16e to 3Dm and demonstrated that while each of them is able to bind to the rRNA transcript independently, their simultaneous binding has a noticeable synergetic effect. Using enzymatic footprinting, we showed that these proteins protect 3Dm against hydrolysis with RNases mainly in the regions homologous to the sites of S7p and S9p binding on the 16S rRNA. At the same time, we found regions that correspond to 16S rRNA fragments distant from the binding sites of the respective homologous prokaryotic proteins. Comparison of these results with the data on 3Dm footprinting in binary complexes with S5e or S16e revealed that each of these proteins affects binding of another one to 3Dm, which is displayed in significant expansion of 3Dm sites protected by the proteins against hydrolysis in the ternary complex. PMID- 19559068 TI - A structure-based investigation on the binding interaction of hydroxylated polycyclic aromatic hydrocarbons with DNA. AB - Binding of polycyclic aromatic hydrocarbons (PAHs) with DNA is one of the key steps in their mutagenic process. In this work, a previously established electrochemical displacement method was utilized to measure the binding constants (K(b)) of 26 hydroxylated PAHs (OH-PAHs) with DNA. Eighteen OH-PAHs induced more than 50% signal reduction in the displacement measurement with calf thymus DNA, and K(b) calculated from the EC(50) value is between 8.3 x 10(4) and 3.0 x 10(5)M(-1). Other eight OH-PAHs induced less than 50% signal reduction. For the latter compounds, EC(50) values were obtained by fitting the displacement curve with a bi-exponential decay function and extrapolating it to 50% signal. K(b) was then calculated, and was found to be in the range of 4.5 x 10(4) and 7.6 x 10(4)M(-1). Measurements with polydG.polydC (pdGC) and polydA.polydT (pdAT) demonstrate that the OH-PAHs have no obvious selectivity toward DNA base pairs. In atomic force microscopy, a clear morphological change of calf thymus DNA from linear type to condensation form was observed after binding with 9 hydroxyfluorene. The change is similar to the one observed with the DNA intercalating electrochemical indicator, suggesting that 9-hydroxyfluorene binds with DNA also by intercalation. An examination of the relationship between the molecular structure characteristics of the 26 OH-PAHs and their DNA-binding affinity revealed that the most positive net atomic charge on a hydrogen atom (q(+)(H)) correlated significantly with K(b) at 0.05 level, with sig. (2-tailed) of 0.015. The correlation suggests that hydrogen bonding may play an important role in DNA/OH-PAH binding interaction. It was further revealed that Connolly accessible area (CAA), Connolly molecular area (CMA), and Connolly solvent excluded volume (CSEV) of the nine hydroxybenzo[a]pyrenes correlated significantly with K(b) at 0.01 or 0.05 level, with sig. (2-tailed) 0.019, 0.012 and 0.009, implying that steric effect might be an important factor in the binding of differently substituted OH-PAH with DNA. PMID- 19559056 TI - Regulation of mitochondrial respiratory chain biogenesis by estrogens/estrogen receptors and physiological, pathological and pharmacological implications. AB - There has been increasing evidence pointing to the mitochondrial respiratory chain (MRC) as a novel and important target for the actions of 17beta-estradiol (E(2)) and estrogen receptors (ER) in a number of cell types and tissues that have high demands for mitochondrial energy metabolism. This novel E(2)-mediated mitochondrial pathway involves the cooperation of both nuclear and mitochondrial ERalpha and ERbeta and their co-activators on the coordinate regulation of both nuclear DNA- and mitochondrial DNA-encoded genes for MRC proteins. In this paper, we have: 1) comprehensively reviewed studies that reveal a novel role of estrogens and ERs in the regulation of MRC biogenesis; 2) discussed their physiological, pathological and pharmacological implications in the control of cell proliferation and apoptosis in relation to estrogen-mediated carcinogenesis, anti-cancer drug resistance in human breast cancer cells, neuroprotection for Alzheimer's disease and Parkinson's disease in brain, cardiovascular protection in human heart and their beneficial effects in lens physiology related to cataract in the eye; and 3) pointed out new research directions to address the key questions in this important and newly emerging area. We also suggest a novel conceptual approach that will contribute to innovative regimens for the prevention or treatment of a wide variety of medical complications based on E(2)/ER-mediated MRC biogenesis pathway. PMID- 19559070 TI - Age-related prefrontal over-recruitment in semantic memory retrieval: Evidence from successful face naming and the tip-of-the-tongue state. AB - Studies that have attempted to determine the effects of aging on the brain neural sources of memory retrieval have reported two contrasting age effects: under recruitment and over-recruitment of several prefrontal areas. However, the causes for these effects are still a matter of debate. In order to study the underlying factors that cause the effects, we compared brain activation in young and older adults, in a successful word retrieval condition, and a failed word retrieval condition: the tip-of-the-tongue state. For this, we used the event-related potentials technique and neural source estimation with low-resolution tomographies. The results showed that the older adults did not display under recruitment in any brain area in comparison with the young adults. However, they displayed additional prefrontal activation that varied depending on the processing stage and the condition, which supports the hypothesis of selective over-recruitment in older adults. PMID- 19559071 TI - JAK2V617F allele burden and thrombosis: a direct comparison in essential thrombocythemia and polycythemia vera. AB - OBJECTIVE: A direct comparison of the incidence and risk factors of major thrombosis in essential thrombocythemia (ET) and polycythemia vera (PV) according to their respective JAK2V617F allele burden is the object of this study. MATERIALS AND METHODS: We compared the rate (%/patients/year) of major thrombosis in 867 ET patients (57% JAK2V617F) with that of 415 PV patients (all JAK2V617F) and examined risk factors. RESULTS: Patients with ET wild-type, ET V617F, and PV showed a rate of thrombosis of 1.4%, 2.1%, and 2.7%/patients/year, respectively. The latter was found to progressively increase according to time of diagnosis. Actuarial probability of arterial and venous thrombosis in the first 5 years of diagnosis was roughly similar in the three groups. While in the subsequent periods, the curves of mutated ET patients diverged from wild-type, and after 10 to 15 years the ET-mutated arm approached PV. CONCLUSION: These findings support the concept of a continuum between ET JAK2 mutated and PV, not only in reference to the hematological phenotype, but also in terms of vascular events. PMID- 19559072 TI - Acute and chronic fentanyl administration causes hyperalgesia independently of opioid receptor activity in mice. AB - Although mu-receptor opioids are clinically important analgesics, they can also paradoxically cause hyperalgesia independently of opioid receptor activity, presumably via the action of neuroexcitatory glucoronide metabolites. However, it is unknown whether the commonly used mu-receptor opioid analgesic fentanyl, which is not subject to glucuronidation, can also induce hyperalgesia independently of opioid receptor activity. Thus, here we examined whether fentanyl increases nociception on the tail-withdrawal test in CD-1 mice concurrently treated with the opioid receptor antagonist naltrexone or in opioid receptor triple knock-out mice lacking mu, delta, and kappa opioid receptors. For both groups, an acute fentanyl bolus dose (0.25mg/kg, s.c.) and continuous fentanyl infusion (cumulative daily dose: 10mg/kg) did not cause analgesia at any time. Instead, fentanyl significantly decreased withdrawal latencies relative to pre-drug values for the next 15-60 min and for six days, respectively. MK-801 blocked and reversed hyperalgesia caused by the acute injection and continuous infusion of fentanyl, respectively, in naltrexone-treated CD-1 mice, indicating the contribution of NMDA receptors to fentanyl hyperalgesia. These data show that the synthetic opioid fentanyl causes hyperalgesia independently of prior or concurrent opioid receptor activity or analgesia. Since the biotransformation of fentanyl does not yield any known pronociceptive metabolites, these data challenge assumptions regarding the role of neuroexcitatory metabolites in opioid induced hyperalgesia. PMID- 19559073 TI - Immuohistochemical markers for pituicyte. AB - GFAP has long been adopted as the specific marker for pituicyte, a special type of astrocyte. GFAP and S100beta are two commonly used astrocyte markers. Their immunoreactivities differ in different regions of the brain. To our knowledge this issue has not been studied in pituicyte. In our preliminary study, we found that antibodies against GFAP and S100beta stained the pituicytes differently. A detailed investigation with both light and electron microscopic techniques was thus conducted in the rat. At light microscopic level, anti-GFAP and anti S100beta stained 66.78% and 86.78% of the pituicytes, respectively. It was found at ultrastructural level that this difference was cell type specific. The parenchymatous pituicytes could be stained with antibodies against both GFAP and S100beta, whereas the fibrous pituicytes were only S100beta-immunoreactive. The functional significance of this cell type specificity remains to be elucidated. PMID- 19559074 TI - Effects of a high-frequency, low-intensity, biphasic conditioning train of TMS pulses on the human motor cortex. AB - Intracortical circuit excitability of the human motor cortex has been studied by measuring effects of some conditioning TMS stimulus on the succeeding test TMS stimulus in the motor cortex, such as short-interval intracortical inhibition (SICI) and intracortical facilitation (ICF). A single-pulse TMS was used as a conditioning stimulus (CS) in these techniques, but a train of several TMS pulses might induce some intracortical changes in the motor cortex more effectively. For nine healthy volunteers, we compared the SICI and ICF induced by a single conditioning biphasic TMS pulse with those induced by a train of 10 biphasic TMS pulses of the same intensity. As a conditioning stimulus, we delivered a subthreshold single biphasic pulse (CS1) or 10, 10-Hz biphasic pulses (CS10) before a suprathreshold monophasic test stimulus at several interstimulus intervals (ISIs) of 3-40 ms over the hand motor area. The CS intensity was 50 100% of the active motor threshold (AMT). We compared the motor cortical excitability after the conditioning stimulus (single pulse or a train of ten pulses) at the intervals for SICI and ICF. A train of ten 10-Hz pulses elicited greater inhibition at short ISIs than a single conditioning pulse did. The facilitation at ISIs around 10 ms corresponding to the ICF was evoked by CS1 only at an intensity of 80% AMT; CS10 evoked no ICF. Furthermore, CS10 evoked MEP inhibition at longer intervals. Results show that a train of high-frequency, low intensity, biphasic TMS pulses can have a strong inhibitory effect on the motor cortex. PMID- 19559075 TI - Effect of cervical spinal cord hemisection on the expression of axon growth markers. AB - To evaluate the plasticity processes occurring in the spared and injured tissue after partial spinal cord injury, we have compared the level of axon growth markers after a C2 cervical hemisection in rats between the contralateral (spared) and ipsilateral (injured) cervical cord using western blotting and immunohistochemical techniques. In the ipsilateral spinal cord 7 days after injury, although GAP-43 levels were increased in the ventral horn caudal to the injury, they were globally decreased in the whole structure (C1-C6). By contrast, in the contralateral intact side 7 days and 1 month after injury, we have found an increase of GAP-43 and betaIII tubulin levels, suggesting that processes of axonal sprouting may occur in the spinal region contralateral to the injury. This increase of GAP-43 in the contralateral spinal cord after cervical hemisection may account, at least partially, to the spontaneous ipsilateral recovery observed after a cervical hemisection. PMID- 19559076 TI - The dose-effectiveness of intranasal VEGF in treatment of experimental stroke. AB - The aim of the present study was to assess the dose-effectiveness of intranasal (IN) vascular endothelial growth factor (VEGF)in the treatment of experimental stroke. Sprague-Dawley rats were randomized into four groups as IN low (100 microg/ml), IN middle (200 microg/ml) and IN high (500 microg/ml) VEGF-treated group, and IN saline-treated group (n=12), given recombinant human VEGF 165 or saline intranasally. Focal cerebral ischemia was induced by transient (90 min) middle cerebral artery occlusion (MCAO) method. Behavioral neurological deficits were assessed 1, 7 and 14 d after the onset of MCAO. Rats were sacrificed at 14 d, the brain sections were stained and an image analysis system was used to calculate the infarct volume. Microvessels were labeled by FITC-dextran and the segment lengths, diameters and number of microvessels were measured by Image Pro Plus Version 6.0 software. Fourteen days post MCAO, infarct volume significantly reduced (P<0.01) in rats which received the middle dose of IN VEGF when compared to IN saline. And middle dose of VEGF significantly improved behavioral recovery (P<0.01). No significant difference in the behavioral recovery and infarct volume was observed between the saline-treated group and the low or high VEGF-treated groups (P>0.05). Compared to IN saline, middle and high doses of VEGF significantly increased the segment length, diameter and number of microvessels (P<0.01). No significant difference in the segment length, diameter and number of microvessels was observed between the IN saline-treated group and the low VEGF treated group (P>0.05). The middle dose of IN VEGF was most effective on reducing infarct volume, improving behavioral recovery and enhancing angiogenesis in stroke brain, which can be used in the following treatments to further evaluate the effect of VEGF. PMID- 19559078 TI - Contribution of in vitro neurotoxicology studies to the elucidation of neurodegenerative processes. AB - Only a small percentage of neurodegenerative diseases like Alzheimer's disease and Parkinson's disease is directly related to familial forms. The etiology of the most abundant, sporadic forms seems to involve both genetic and environmental factors. Environmental compounds are now extensively studied for their possible contribution to neurodegeneration. Chemicals were found which were able to reproduce symptoms of known neurodegenerative diseases, others may either predispose to the onset of neurodegeneration, or exacerbate distinct pathogenic processes of these diseases. In any case, in vitro studies performed with models presenting various degrees of complexity have shown that many environmental compounds have the potential to cause neurodegeneration, through a variety of pathways similar to those described in neurodegenerative diseases. Since the population is exposed to a huge number of potentially neurotoxic compounds, there is an important need for rapid and efficient procedures for hazard evaluation. Xenobiotics elicit a cascade of reactions that, most of the time, involve numerous interactions between the different brain cell types. A reliable in vitro model for the detection of environmental toxins potentially at risk for neurodegenerative diseases should therefore allow maximal cell-cell interactions and multiparametric endpoints determination. The combined use of in vitro models and new analytical approaches using "omics" technologies should help to map toxicity pathways, and advance our understanding of the possible role of xenobiotics in the etiology of neurodegenerative diseases. PMID- 19559079 TI - A new cerebroside from Gynura divaricata. AB - A new cerebroside, 1-O-beta-D-glucopyranosyl-(2S,3S,4R,10E)-2-[(2'R)-2' hydroxyltricosanoyl-amino]-10-octadecene-1,3,4-triol was isolated from the aerial parts of Gynura divaricata DC. PMID- 19559080 TI - Flavonoids from Inula britannica L. inhibit injury-induced neointimal formation by suppressing oxidative-stress generation. AB - AIM OF THE STUDY: We aimed to investigate whether and how the total flavonoid extracts (TFE) from Inula britannica L. block neointimal hyperplasia induced by balloon injury in rats. MATERIALS AND METHODS: Rats were administered orally TFE doses of 12.5, 25 and 50 mg/kg/d by gastric gavage from 3 days before balloon injury to 14 days after the injury. The ratio of intima (I) to media (M) thickness (I/M) in carotid arteries was examined by morphological analyses. The MDA content and SOD activity in plasma were measured. The O(2)(-) production in vascular tissues was detected in situ. The expression of p47(phox) in carotid arteries was analyzed by Western blot analysis and immunohistochemistry. RESULTS: The rats treated with TFE 50 mg/kg/d showed a reduction in neointimal hyperplasia, and the ratio of I/M of balloon injured-carotid arteries was significantly reduced by over 70% after TFE treatment, compared with the injured group. The inhibitory effect of TFE on neointimal hyperplasia was almost consistent with that of atorvastatin, a positive control. The plasma SOD activity was obviously increased by TFE treatment (P<0.01), while plasma MDA production was markedly decreased by TFE treatment (P<0.05). On day 14 after balloon injury, the carotid arteries showed an increase in O(2)(-) production that was most evident in the neointimal and medial layer of the vessel. Thus, TFE significantly inhibited injury-induced O(2)(-) production and p47(phox) expression in carotid arteries. CONCLUSION: Our results suggest that TFE inhibit the neointimal hyperplasia after balloon injury, at least partly, by suppressing oxidative stress generation. PMID- 19559077 TI - Estrogen receptor beta regulates the expression of tryptophan-hydroxylase 2 mRNA within serotonergic neurons of the rat dorsal raphe nuclei. AB - Dysfunctions of the brain 5-HT system are often associated with affective disorders, such as depression. The raphe nuclei target the limbic system and most forebrain areas and constitute the main source of 5-HT in the brain. All 5-HT neurons express tryptophan hydroxylase-2 (TPH2), the brain specific, rate limiting enzyme for 5-HT synthesis. Estrogen receptor (ER) beta agonists have been shown to attenuate anxiety- and despair-like behaviors in rodent models. Therefore, we tested the hypothesis that ER beta may contribute to the regulation of gene expression in 5-HT neurons of the dorsal raphe nuclei (DRN) by examining the effects of systemic and local application of the selective ER beta agonist diarylpropionitrile (DPN) on tph2 mRNA expression. Ovariectomized (OVX) female rats were injected s.c. with DPN or vehicle once daily for 8 days. In situ hybridization revealed that systemic DPN-treatment elevated basal tph2 mRNA expression in the caudal and mid-dorsal DRN. Behavioral testing of all animals in the open field (OF) and on the elevated plus maze (EPM) on days 6 and 7 of treatment confirmed the anxiolytic nature of ER beta activation. Another cohort of female OVX rats was stereotaxically implanted bilaterally with hormone containing wax pellets flanking the DRN. Pellets contained 17-beta-estradiol (E), DPN, or no hormone. Both DPN and E significantly enhanced tph2 mRNA expression in the mid-dorsal DRN. DPN also increased tph2 mRNA in the caudal DRN. DPN- and E treated rats displayed a more active stress-coping behavior in the forced-swim test (FST). No behavioral differences were found in the OF or on the EPM. These data indicate that ER beta acts at the level of the rat DRN to modulate tph2 mRNA expression and thereby influence 5-HT synthesis in DRN subregions. Our results also suggest that local activation of ER beta neurons in the DRN may be sufficient to decrease despair-like behavior, but not anxiolytic behaviors. PMID- 19559081 TI - In-hospital levels of C-reactive protein and IL-6 predict post-operative depressive symptoms among patients undergoing total knee replacement surgery. AB - Behavioral changes observed following immune system activation are similar to many of the hallmark symptoms of major depressive disorder (MDD), including appetite change, lethargy, fatigue, negative mood and anhedonia. Acute phase proteins, such as interleukin-6 (IL-6) and C-reactive protein (CRP) have been implicated in the production of sickness behavior, and research has revealed significant differences in the levels of these acute phase proteins between depressed and non-depressed individuals. The current study examined whether early post-operative IL-6 and CRP levels predicted subsequent depressive symptoms in 110 patients undergoing total knee replacement surgery (TKR). In-hospital levels of IL-6 and CRP predicted depressive symptoms at three-months following surgery, as indicated by significant main effects and a significant interaction term. Specifically, lower levels of in-hospital CRP and higher levels of IL-6 in hospital predicted more depressive symptoms three-months following surgery. The finding that levels of acute phase proteins soon after surgery predict subsequent depressive symptoms, if replicated, extends prior research on the relationships between IL-6, CRP, and depression. Further, this predictive relationship suggests the possibility of early identification of individuals at risk for the subsequent development of post-operative depression. PMID- 19559082 TI - Aryl hydrocarbon receptor-dependent induction of the NADPH oxidase subunit NCF1/p47 phox expression leading to priming of human macrophage oxidative burst. AB - Polycyclic aromatic hydrocarbons such as benzo(a)pyrene (BaP) are toxic environmental contaminants known to regulate gene expression through activation of the aryl hydrocarbon receptor (AhR). In the present study, we demonstrated that acute treatment by BaP markedly increased expression of the NADPH oxidase subunit gene neutrophil cytosolic factor 1 (NCF1)/p47(phox) in primary human macrophages; NCF1 was similarly up-regulated in alveolar macrophages from BaP instilled rats. NCF1 induction in BaP-treated human macrophages was prevented by targeting AhR, through its chemical inhibition or small interference RNA-mediated down-modulation of its expression. BaP moreover induced activity of the NCF1 promoter sequence, containing a consensus AhR-related xenobiotic-responsive element (XRE), and electrophoretic mobility shift assays and chromatin immunoprecipitation experiments indicated that BaP-triggered binding of AhR to this XRE. Finally, we showed that BaP exposure resulted in p47(phox) protein translocation to the plasma membrane and in potentiation of phorbol myristate acetate (PMA)-induced superoxide anion production in macrophages. This BaP priming effect toward NADPH oxidase activity was inhibited by the NADPH oxidase specific inhibitor apocynin and the chemical AhR inhibitor alpha-naphtoflavone. These results indicated that BaP induced NCF1/p47(phox) expression and subsequently enhanced superoxide anion production in PMA-treated human macrophages, in an AhR-dependent manner; such an NCF1/NADPH oxidase regulation by polycyclic aromatic hydrocarbons may participate in deleterious effects toward human health triggered by these environmental contaminants, including atherosclerosis and smoking-related diseases. PMID- 19559083 TI - Neural progenitor cells as models for high-throughput screens of developmental neurotoxicity: state of the science. AB - In vitro, high-throughput methods have been widely recommended as an approach to screen chemicals for the potential to cause developmental neurotoxicity and prioritize them for additional testing. The choice of cellular models for such an approach will have important ramifications for the accuracy, predictivity and sensitivity of the screening assays. In recent years neuroprogenitor cells from rodents and humans have become more widely available and may offer useful models having advantages over primary neuronal cultures and/or transformed cell lines. To date, these models have been utilized in only a limited number of toxicity studies. This review summarizes the state of the science regarding stem and neuroprogenitor models that could be used for screening assays, provides researchers in this field with examples of how these cells have been utilized to date, and discusses the advantages, limitations and knowledge gaps regarding these models. Data are available from both rodent and human stem and neuroprogenitor cell models that indicate that these models will be a valid and useful tool for developmental neurotoxicity testing. Full potential of these models will only be achieved following advances in neurobiology that elucidate differentiation pathways more clearly, and following further evaluation of larger sets of developmentally neurotoxic and non-toxic chemicals to define the sensitivity and predictivity of assays based on stem or progenitor cell models. PMID- 19559085 TI - Comparison of PC12 and cerebellar granule cell cultures for evaluating neurite outgrowth using high content analysis. AB - Development of high-throughput assays for chemical screening and hazard identification is a pressing priority worldwide. One approach uses in vitro, cell based assays which recapitulate biological events observed in vivo. Neurite outgrowth is one such critical cellular process underlying nervous system development that can be quantified using automated microscopy and image analysis (high content analysis). The present study characterized and compared the PC-12 cell line (NS-1) and primary cultures of cerebellar granular cells (CGC), as models for assessing chemical effects on neurite outgrowth. High content analysis of neurite outgrowth was performed using the Cellomics ArrayScan V(Ti) automated epifluorescent imaging system to acquire and analyze images of beta-tubulin immunostained cells in 96-well plates. Cell viability was assessed using the CellTiter-Glo assay. Culture of NS-1 or CGC in nerve growth factor or serum respectively, rapidly induced neurite outgrowth that increased over four days in vitro. Seven compounds previously shown to affect neurite outgrowth in vitro were tested in both models for changes in total neurite length and cell viability. In NS-1 cells, four chemicals (PKC inhibitor Bis-I, MEK inhibitor U0126, trans Retinoic acid, methylmercury) inhibited neurite outgrowth, while lead, amphetamine and valproic acid had no effect. In CGC, five chemicals inhibited neurite outgrowth (Bis-I, U0126, lead, methylmercury, and amphetamine), while trans-Retinoic acid decreased cell viability but not neurite outgrowth. Valproic acid was without effect. The sensitivity of the two models was chemical specific: NS-1 cells were more sensitive to Bis-I, methylmercury and trans-Retinoic acid, while CGC were more sensitive to U0126, lead, and amphetamine. For every chemical (except trans-Retinoic acid), neurite outgrowth was equal to or more sensitive than cell viability. In comparison, out of seven chemicals without prior evidence for effects on neurite outgrowth, only one decreased neurite outgrowth (diphenhydramine in CGC). These findings demonstrate that the effects of chemicals on neurite outgrowth may be cell type specific. PMID- 19559086 TI - Measuring charge and mass distributions in dry powder inhalers using the electrical Next Generation Impactor (eNGI). AB - The electrical Next Generation Impactor (eNGI) was assessed against the electrical low-pressure impactor (ELPI) and next generation impactor (NGI) for its capability to characterise particle size and electrostatic charge properties of dry powder inhaler (DPI) formulations. Following assessment, the relationship between inhalational air flow rate and drug powder charge was explored using the eNGI. At a vacuum flow rate of 30L/min, doses of Pulmicort (budesonide 400 microg) and Bricanyl (terbutaline 500 microg) were dispersed into the ELPI, NGI and eNGI, from which particle size profiles and charge profiles were ascertained. Further doses of Pulmicort and Bricanyl were fired into the eNGI at vacuum air flow rates of 45, 60, 75 and 90 L/min, and the resultant size and charge profiles were determined. Particle size profiles at 30 L/min were found to be comparable between the NGI and eNGI, while charge profiles were comparable between the eNGI and ELPI. As air flow rate increased from 30 to 90 L/min, in vitro aerosol performance improved before reaching a peak at 45 L/min (Pulmicort) and 60 L/min (Bricanyl). Net charge also increased with flow rate, the cause of which may be a combination of increased turbulence and aerosol performance. This study demonstrates that the eNGI is capable of electrostatic and particle size characterization of commercial drug-only DPI products. PMID- 19559084 TI - Drosophotoxicology: the growing potential for Drosophila in neurotoxicology. AB - Understanding neurotoxic mechanisms is a challenge of deciphering which genes and gene products in the developing or mature nervous system are targeted for disruption by chemicals we encounter in our environment. Our understanding of nervous system development and physiology is highly advanced due in large part to studies conducted in simple non-mammalian models. The paucity of toxicological data for the more than 80,000 chemicals in commercial use today, and the approximately 2000 new chemicals introduced each year, makes development of sensitive and rapid assays to screen for neurotoxicity paramount. In this article I advocate the use of Drosophila in the modern regimen of toxicological testing, emphasizing its unique attributes for assaying neurodevelopment and behavior. Features of the Drosophila model are reviewed and a generalized overall scheme for its use in toxicology is presented. Examples of where the fly has proven fruitful in evaluating common toxicants in our environment are also highlighted. Attention is drawn to three areas where development and application of the fly model might benefit toxicology the most: 1) optimizing sensitive endpoints for pathway-specific screening, 2) accommodating high throughput demands for analysis of chemical toxicant libraries, 3) optimizing genetic and molecular protocols for more rapid identification toxicant-by-gene interactions. While there are shortcomings in the Drosophila model, which exclude it from effective toxicological testing in certain arenas, conservation of fundamental cellular and developmental mechanisms between flies and man is extensive enough to warrant a central role for the Drosophila model in toxicological testing of today. PMID- 19559087 TI - Is nicotine a key player or spectator in the induction and progression of cardiovascular disorders? AB - Cigarette smoking is common in societies worldwide and a growing body of evidence suggests that chronic cigarette smoking may affect the structure and function of cardiovascular system. The chronic exposure to high levels of nicotine, a major component of cigarette smoking, has been observed to play a pathogenic role in the induction and progression of cardiovascular disorders including cardiomyopathy and peripheral vascular disease. Nicotine alters the function of vascular endothelium, initiates the adhesion cascade and stimulates the vascular inflammatory events to induce atherosclerosis and hypertension. Moreover, nicotine has been noted to induce direct coronary spasm and ischemia, which develop coronary artery disease and myocardial infarction. In addition, nicotine stimulates the excessive release of impulses from sinoatrial node that may account for the induction of cardiac arrhythmia. The present review critically discussed the possible detrimental role of chronic nicotine exposure in cardiac and vascular endothelial dysfunction. Moreover, the signaling mechanisms involved in the pathogenesis of nicotine exposure-induced cardiovascular dysfunction have been discussed. In addition, the pharmacological interventions to ameliorate chronic nicotine exposure-induced cardiovascular abnormalities have been delineated. PMID- 19559090 TI - Stable isotope pulse-chase monitored by quantitative mass spectrometry applied to E. coli 30S ribosome assembly kinetics. AB - Stable isotope mass spectrometry has become a widespread tool in quantitative biology. Pulse-chase monitored by quantitative mass spectrometry (PC/QMS) is a recently developed stable isotope approach that provides a powerful means of studying the in vitro self-assembly kinetics of macromolecular complexes. This method has been applied to the Escherichia coli 30S ribosomal subunit, but could be applied to any stable self-assembling complex that can be reconstituted from its component parts and purified from a mixture of components and complex. The binding rates of 18 out of the 20 ribosomal proteins have been measured at several temperatures using PC/QMS. Here, PC/QMS experiments on 30S ribosomal subunit assembly are described, and the potential application of the method to other complexes is discussed. A variation on the PC/QMS experiment is introduced that enables measurement of kinetic cooperativity between proteins. In addition, several related approaches to stable isotope labeling and quantitative mass spectrometry data analysis are compared and contrasted. PMID- 19559091 TI - The role of actions in auditory object discrimination. AB - Action representations can interact with object recognition processes. For example, so-called mirror neurons respond both when performing an action and when seeing or hearing such actions. Investigations of auditory object processing have largely focused on categorical discrimination, which begins within the initial 100 ms post-stimulus onset and subsequently engages distinct cortical networks. Whether action representations themselves contribute to auditory object recognition and the precise kinds of actions recruiting the auditory-visual mirror neuron system remain poorly understood. We applied electrical neuroimaging analyses to auditory evoked potentials (AEPs) in response to sounds of man-made objects that were further subdivided between sounds conveying a socio-functional context and typically cuing a responsive action by the listener (e.g. a ringing telephone) and those that are not linked to such a context and do not typically elicit responsive actions (e.g. notes on a piano). This distinction was validated psychophysically by a separate cohort of listeners. Beginning approximately 300 ms, responses to such context-related sounds significantly differed from context free sounds both in the strength and topography of the electric field. This latency is >200 ms subsequent to general categorical discrimination. Additionally, such topographic differences indicate that sounds of different action sub-types engage distinct configurations of intracranial generators. Statistical analysis of source estimations identified differential activity within premotor and inferior (pre)frontal regions (Brodmann's areas (BA) 6, BA8, and BA45/46/47) in response to sounds of actions typically cuing a responsive action. We discuss our results in terms of a spatio-temporal model of auditory object processing and the interplay between semantic and action representations. PMID- 19559088 TI - Exploring ribozyme conformational changes with X-ray crystallography. AB - Relating three-dimensional fold to function is a central challenge in RNA structural biology. Toward this goal, X-ray crystallography has long been considered the "gold standard" for structure determinations at atomic resolution, although NMR spectroscopy has become a powerhouse in this arena as well. In the area of dynamics, NMR remains the dominant technique to probe the magnitude and timescales of molecular motion. Although the latter area remains largely unassailable by conventional crystallographic methods, inroads have been made on proteins using Laue radiation on timescales of ms to ns. Proposed 'fourth generation' radiation sources, such as free-electron X-ray lasers, promise ps- to fs-timescale resolution, and credible evidence is emerging that supports the feasibility of single molecule imaging. At present however, the preponderance of RNA structural information has been derived from timescale and motion insensitive crystallographic techniques. Importantly, developments in computing, automation and high-flux synchrotron sources have propelled the rapidity of 'conventional' RNA crystal structure determinations to timeframes of hours once a suitable set of phases is obtained. With a sufficient number of crystal structures, it is possible to create a structural ensemble that can provide insight into global and local molecular motion characteristics that are relevant to biological function. Here we describe techniques to explore conformational changes in the hairpin ribozyme, a representative non-protein-coding RNA catalyst. The approaches discussed include: (i) construct choice and design using prior knowledge to improve X-ray diffraction; (ii) recognition of long-range conformational changes and (iii) use of single-base or single-atom changes to create ensembles. The methods are broadly applicable to other RNA systems. PMID- 19559089 TI - In vivo manipulation of gene expression in non-human primates using lentiviral vectors as delivery vehicles. AB - Non-human primates (NHPs) are an invaluable resource for the study of genetic regulation of disease mechanisms. The main disadvantage of using NHPs as a preclinical model of human disease is the difficulty of manipulating the monkey genome using conventional gene modifying strategies. Lentiviruses offer the possibility of circumventing this difficulty because they can infect and transduce either dividing or nondividing cells, without producing an immune response. In addition, lentiviruses can permanently integrate into the genome of host cells, and are able to maintain long-term expression. In this article we describe the lentiviral vectors that we use to both express transgenes and suppress expression of endogenous genes via RNA interference (RNAi) in NHPs. We also discuss the safety features of currently available vectors that are especially important when lentiviral vectors are used in a species as closely related to humans as NHPs. Finally, we describe in detail the lentiviral vector production protocol we use and provide examples of how the vector can be employed to target peripheral tissues and the brain. PMID- 19559092 TI - Molecular taxonomy and species delimitation in Andean Schistocerca (Orthoptera: Acrididae). AB - The acridian genus Schistocerca comprises about 50 species which are endemic to the New World, except the Old World locust S. gregaria. Their morphological identification is rendered difficult by phase polyphenism, geographical overlap due to migrations or swarming, the difficulty to easily differentiate genitalia and the occurrence of interspecific hybrids. The three species reported from Peru include the swarming species S. interrita, a pest that can be recognized only by taxonomists. We show that it can be unambiguously identified using a mitochondrial DNA fragment known to have barcoding properties in this genus. We used several methods to delimitate Peruvian species. While S. interrita and S. pallens were well characterized, S. piceifrons peruviana was split into several taxa by a phylogeny-based method, whereas a combination of population genetics methods led one to identify only the three nominal species. A tentative reconstruction of the species history shows that several populations of S. piceifrons peruviana have recently increased in number, while exchanging some migrants, whereas an isolated population at the northern margin of the species range is substantially differentiated while exchanging no migrants with the others. This complex history has resulted in an atypical lineage pattern that appears to have confounded the standard assumptions underlying available species delimitation methods. Because of its behavioral property which tends to keep it panmictic, the identification of the swarming S. interrita remained unaffected. PMID- 19559093 TI - Genomic interactions: chromatin loops and gene meeting points in transcriptional regulation. AB - The chromosome conformation capture (3C) technique and its genome-wide applications ('4C') have identified a plethora of distal DNA sequences that are frequently in close spatial proximity. In many cases, these have been correlated with transcriptional regulation of the interacting genes, but the functional significance of many of the extreme long-range and interchromosomal interactions remains unclear. This review summarises our current understanding of how chromatin conformation can impinge on gene expression, the major questions that need to be addressed to understand this more fully, and how these questions may be answered in the near future. PMID- 19559094 TI - AFLP genotyping of Candida metapsilosis clinical isolates: evidence for recombination. AB - In a collection of 395 independent clinical isolates classified as Candida parapsilosis on a biochemical profile basis, 20 Candida metapsilosis strains were identified by molecular tests with an isolation frequency of 5%. Isolates were screened for their susceptibility to conventionally used antifungals and for virulence determinants, such as biofilm formation and protease production. Molecular characterization of C. metapsilosis independent isolates by amplified fragment length polymorphism (AFLP) revealed a high percentage of polymorphic bands. Statistical analysis of the pairwise genetic distances and bootstrapping revealed that recombination occurs and significantly contributes to C. metapsilosis genetic population variability. No association between specific AFLP markers and drug resistance or other phenotypes was observed. PMID- 19559095 TI - Pharmacological inhibition of inducible nitric oxide synthase attenuates the development of seizures in mice. AB - The present study has been designed to pharmacologically expound the significance of inducible nitric oxide synthase in the pathophysiological progression of seizures using mouse models of chemically induced kindled epilepsy and status epilepticus induced spontaneous recurrent seizures. Pentylenetetrazole (40 mg kg( 1)) (PTZ) administration every second day for a period of 15 days was used to elicit kindled seizure activity in mice. Severity of kindled seizures was assessed in terms of a composite kindled seizure severity score (KSSS). Pilocarpine (100 mg kg(-1)) was injected every 20 min until the onset of status epilepticus. A spontaneous recurrent seizure severity score (SRSSS) was recorded as a measure of quantitative assessment of the progressive development of spontaneous recurrent seizures induced after pilocarpine status epilepticus. Sub acute PTZ administration induced the development of severe form of kindled seizures in mice. Further, pharmacological status epilepticus elicited a progressive evolution of spontaneous recurrent seizures in the animals. However, treatment of aminoguanidine, a relatively selective inhibitor of inducible nitric oxide synthase, markedly and dose dependently suppressed the development of both PTZ induced kindled seizures as well as pilocarpine induced spontaneous recurrent seizures. Therefore inducible nitric oxide synthase may be implicated in the development of seizures. PMID- 19559096 TI - Induction of vitellogenin production in male tilapia (Oreochromis mossambicus) by commercial fish diets. AB - Mozambique tilapia, (Oreochromis mossambicus), are a euryhaline teleost and an important biological model species. Captive male tilapia frequently have high levels of the estrogen-induced yolk precursor protein vitellogenin (Vg), a common indicator of exposure to estrogenic compounds. Sex steroids are found in commercial fish diets, but relatively few studies have examined the relationship between commercial diets and Vg production. In a fasting experiment to ascertain a dietary role in male Vg production, plasma Vg was reduced to negligible levels after 2 weeks of fasting, while no change in estrogen receptor (ER) expression was seen. When male tilapia were fed a squid-based diet that replaced the commercial trout diet, plasma Vg was reduced to undetectable levels over 40 days, concomitant with significant reductions in hepatic expression of Vgs A, B, and C, and ERbeta, compared with control fish fed commercial trout diet. Female tilapia fed the squid-based for 20 days had no change in these parameters. When male tilapia were fed a defined, soy-based diet, plasma Vg reduced to 20% of levels in fish given either commercial trout diet or a defined, fishmeal-based diet. Overall, results from these studies suggest that estrogens in a commercial trout diet induce vitellogenin production by increasing expression of Vg, but not ER genes in male tilapia. PMID- 19559097 TI - Purification and partial characterization of a myofibril-bound serine protease from ostrich skeletal muscle. AB - A myofibril-bound serine protease (MBSP) was partially purified from ostrich (Struthio camelus) skeletal muscle. MBSP was dissociated from the myofibrillar fraction by ethylene glycol treatment at pH 8.5, followed by partial purification via Toyopearl Super Q 650 S and p-aminobenzamidine column chromatographies. Ostrich MBSP revealed a major protein band of approximately 21 kDa on SDS-PAGE, showing proteolytic activity after casein zymography. Optima pH and temperature of ostrich MBSP were 8 and 40 degrees C, respectively. Substrate specificity analysis revealed that the enzyme cleaved synthetic fluorogenic substrates at the carboxyl side of arginine residues. Kinetic parameters (K(m) and V(max) values) were calculated from Lineweaver-Burk plots. The kinetic characteristics of ostrich MBSP were compared to values obtained for commercial bovine trypsin in this study, as well as those obtained for MBSP from mouse and various fish species. The results suggest that ostrich MBSP is a tryptic-like serine protease. Ostrich MBSP exhibited low sequence identity to commercial bovine trypsin (44%), MBSP from lizard fish skeletal muscle (33%) and trypsinogen from ostrich pancreas (22%). PMID- 19559098 TI - A novel and simple method of screening compounds for interaction with DNA: A validation study. AB - We report the development of a simple, cost-effective assay for detecting compounds that have the ability to interact with and modify DNA. Potential uses for the assay lie in the areas of early genotoxicity testing of drug candidates, anticancer and antibiotic drug discovery, environmental monitoring and testing in the food, beverage and cosmetics industries. At present the assay has been used to assess direct-acting compounds only and it is yet to be established whether the assay is compatible with bio-activation. The methodology is based on the oxidative reaction of potassium permanganate with pyrimidine bases, which have become perturbed and more reactive by the agent under test. Results are recorded by use of UV/vis spectroscopy. The adaptation to a multi-well plate format provides the capacity for high throughput utilizing small amounts of compounds. Over 100 compounds, comprising different classes of DNA-binding chemicals as well as non-binding controls, have been put through the assay and the results compared with existing genotoxicity testing data from other methods. The assay has shown to be predictive of the results of other genotoxicity testing methods. We have found that the method is overall predictive of 71% of Ames bacterial reverse mutation test results (where data are given) encompassing both negative and positive results. PMID- 19559100 TI - Study of oxidative DNA damage in TK6 human lymphoblastoid cells by use of the in vitro micronucleus test: Determination of No-Observed-Effect Levels. AB - The existence of thresholds for indirect DNA-damaging agents has been widely accepted in the last few years. In contrast, DNA-reactive agents have been assumed to have a non-threshold mode of action, as they directly induce DNA lesions that have the potential to be converted into mutations. However, this does not take into account protective factors acting to reduce or repair genotoxic damage. Among the compounds acting through possible threshold mechanisms, some of them induce DNA damage by oxidative stress. In this context, the aim of our study was to investigate the dose-response relationship of well known DNA-oxidizing agents acting through different mechanisms of oxidative stress, viz. potassium bromate, bleomycin and hydrogen peroxide (by the action of glucose oxidase) by assessing the induction of chromosomal damage using the in vitro micronucleus test (MNT) on the human lymphoblastoid cell line TK6. In order to provide a first characterization of their genotoxic mechanism, two treatment schedules were applied. Cells received both short-term treatment followed by a recovery time (1 + 23 h, 2 + 22 h, 3 + 21 h or 6 + 18 h) and long-term treatment (24h continually). Our results show interesting non-linear dose-effect relationships starting with a range of non-mutagenic very low doses allowing the determination of a No-Observed-Effect Level (NOEL) and going step-wise up to higher doses. After a short exposure, three different plateaus were observed suggesting complex activations and interactions of different cellular mechanisms whose nature and efficiency were dose-dependent. In contrast, after a long treatment, the dose-response curves were different depending on the test compound investigated. Therefore, the in vitro MNT seems to be an appropriate predictive test to establish the NOEL(s) of DNA-oxidizing agents. In order to confirm and to determine the origin of the different cellular step-wise responses observed, additional mechanistic studies would be required, especially by means of other genotoxicity endpoints and gene-expression profiling. PMID- 19559099 TI - Mustard gas exposure and carcinogenesis of lung. AB - Sulfur mustard (SM), also known as mustard gas, is an alkylating compound used as a chemical weapon in World War I and by Iraqi forces against Iranians and indigenous Iraqi Kurds during the Iran-Iraq War of the 1980s. Although SM is a proven carcinogen there are conflicting views regarding the carcinogenicity of a single exposure. The present study characterizes lung cancers formed in mustard gas victims from the Iran-Iraq War. METHODS AND MATERIALS: Demographic information and tumor specimens were collected from 20 Iranian male lung cancer patients with single high-dose SM exposures during the Iran-Iraq War. Formalin fixed, paraffin-embedded lung cancers were analyzed by immunohistochemistry for p53 protein. In addition, DNA was extracted from the tissues, PCR amplified and sequenced to identify mutations in the p53 and KRAS genes associated with SM exposure. RESULTS: A relatively early age of lung cancer onset (ranging from 28 to 73 with a mean of 48) in mustard gas victims, particularly those in the non smoking population (mean age of 40.7), may be an indication of a unique etiology for these cancers. Seven of the 20 patients developed lung cancer before the age of 40. Five of 16 cancers from which DNA sequence data was obtainable provided information on eight p53 mutations (within exons 5-8). These mutations were predominately G to A transitions; a mutation consistent with the DNA lesion caused by SM. Two of the lung cancers had multiple p53 point mutations, similar to results obtained from factory workers chronically exposed to mustard agent. No mutations were detected in the KRAS gene. DISCUSSION: The distinguishing characteristics of lung carcinogenesis in these mustard gas victims suggest that a single exposure may increase the risk of lung cancer development in some individuals. PMID- 19559101 TI - Aloe-emodin-induced DNA fragmentation in the mouse in vivo comet assay. AB - Aloe-emodin (AE) and derivatives may be present as undesired components co extracted during extraction of plants containing anthraquinonic derivatives for preparation of diacetylrhein. AE is a well-known in vitro mutagen, but up to now it failed to induce any clear in vivo genotoxic activity in the chromosome aberration assay in rat bone marrow or the in vivo/in vitro UDS test in liver. However, the two target organs noted during rodent carcinogenicity studies with danthron and emodin, two other well-known anthraquinone derivatives, are the colon and the kidney. Therefore, the choice of the organs for testing the genotoxicity of AE, i.e. bone marrow and liver, may be considered inadequate to demonstrate a possible in vivo genotoxic activity. In this context, the in vivo mouse comet assay was performed on both isolated kidney and colon cells in order to demonstrate a possible organospecific genotoxicity after oral administration of AE. Concurrently, the Ames test and the in vitro micronucleus assay with TK6 human lymphoblastoid cells were performed in their microscale version both with S9 from Aroclor 1254-induced liver or kidney, and without S9. AE induced primary DNA damage in the liver and in the kidney as observed between 3 and 6h after two oral administrations at 500, 1000 and 2000mg/kg bw, underlining an in vivo genotoxic mechanism of action. Furthermore, AE induced a clear genotoxic activity both in the Salmonella typhimurium strains TA1537 and TA98 and in the in vitro micronucleus assay in the absence as well as in the presence of metabolic activation. As no significant variation in the genotoxic activity of AE was noted when using either liver or kidney S9-mix, it seems that no quantitatively and/or qualitatively specific renal metabolism occurs. The kidney may be a target organ of AE as it is the major route of excretion. Under such conditions the separation of AE components should take place and the residual content of undesired AE derivatives should be made as low as reasonably achievable. AE present in plant extracts should be considered as an in vivo genotoxin and this property should be taken into account in the risk assessment for human exposure. PMID- 19559102 TI - Spermatozoon ultrastructure of Aponurus laguncula (Digenea: Lecithasteridae), a parasite of Aluterus monoceros (Pisces: Teleostei). AB - The mature spermatozoon of Aponurus laguncula, a parasite of the unicorn leatherjacket Aluterus monoceros, was studied by transmission electron microscopy. The spermatozoon possesses 2 axonemes of the 9+"1" trepaxonematan pattern, attachment zones, a nucleus, a mitochondrion, external ornamentation of the plasma membrane and cortical microtubules. The major features are the presence of: 1) external ornamentation in the anterior part of the spermatozoon not associated with cortical microtubules; 2) one mitochondrion; and 3) cortical microtubules arranged as a single field in the ventral side. The maximum number of microtubules is in the nuclear region. The extremities of the axonemes are characterized by the disappearance of the central core and the presence of microtubule doublets or singlets. This study is the first undertaken with a member of the Lecithasteridae and exemplifies the sperm ultrastructure for the superfamily Hemiuroidea. PMID- 19559103 TI - Analysis of HIV-1 pol sequences from Panama: identification of phylogenetic clusters within subtype B and detection of antiretroviral drug resistance mutations. AB - We report the first study to examine phylogenetic relationships and drug resistance mutations in HIV-1 pol sequences from Panama. For this study, we used plasma RNA samples from 135 HIV-1-infected subjects from Panama, of which 82 (61%) had AIDS and 53 (39%) were asymptomatic drug-naive individuals. Phylogenetic analyses revealed that 133 (98%) subjects were infected with subtype B viruses and 2 AIDS patients harboured recombinant viruses, CRF02_AG/A3 and CRF12_BF/B, respectively. Using a Bayesian phylogeny inference method, 5 strongly supported clusters of > or =5 sequences, designated B-PA1 to B-PA5, were identified within subtype B, together comprising 87 (65.4%) subtype B viruses. Cluster B-PA1 (n=42) was significantly associated with East Panama and clusters B PA2 (n=15) and B-PA4 (n=10) were associated with West Panama. A Bayesian coalescent analysis indicated that the most recent common ancestors of the four largest clusters were dated in the 1980s and that of the fifth in the early 1990s. The analysis of antiretroviral drug resistance-associated mutations revealed that 8 (9.7%) AIDS patients, all of them antiretroviral drug experienced, harboured mutations conferring high or intermediate resistance levels to antiretroviral drugs. No drug resistance mutations were detected among the asymptomatic drug-naive individuals. In conclusion, the phylogenetic analysis of HIV-1 pol sequences from Panama reveals that a majority of subtype B viruses, which are predominant in Panama, branch within well supported intrasubtype phylogenetic clusters, most of them originating in the 1980s, and some with geographical associations, which may reflect either multiple HIV-1 subtype B introductions or the existence of local transmission networks originating in the early epidemic in Panama. PMID- 19559105 TI - Effects of obesity on breathing pattern, ventilatory neural drive and mechanics. AB - The purpose of this study was to assess whether obesity induces changes in breathing pattern and ventilatory neural drive and mechanics. Measurements performed in 34 male obese subjects (BMI, 39+/-6 kg/m(2)) and 18 controls (BMI, 23+/-3 kg/m(2)) included anthropometric parameters, spirometry, breathing patterns, mouth occlusion pressure, maximal inspiratory pressure and work of breathing. The results show that spirometric flow (FEV(1)% pred, FVC% pred) and maximal inspiratory pressure (P(Imax)) were significantly lowers (p<0.001) in obese subjects compared to controls. The (fR/VT) ratio was higher in obese subjects than in controls (p<0.001). The increase in (fR/VT) was associated with an increase in the ratio of mean inspiratory pressure to maximal inspiratory pressure (P(I)/P(Imax)) and the duty cycle (T(I)/T(TOT)) (p<0.001). The energy cost of breathing (W(rest)/W(crit)), which reflects the oxygen consumed by the respiratory muscle was greater in obese subject than in controls (p<0.001) inducing an increase in the effective inspiratory impedance on the respiratory muscles. It is concluded that obese subjects show impairment in breathing pattern and respiratory mechanics as assessed by rapid shallow breathing leading to ventilatory failure. PMID- 19559104 TI - Bayesian coalescent inference reveals high evolutionary rates and expansion of Norovirus populations. AB - Noroviruses (NoV) are a leading cause of outbreaks of nonbacterial acute gastroenteritis in humans worldwide and have become an important cause of hospitalization of children in South America. NoV belong to the family Caliciviridae and are non-enveloped single stranded, positive sense, RNA viruses. NoV of genotype GII/4 have emerged worldwide, causing four epidemic seasons of viral gastroenteritis during which four novel variants emerged. Despite the importance of NoV outbreaks, little is known about the evolutionary rates, viral spread and population dynamics of NoV populations. In order to gain insight into these matters, a Bayesian Markov chain Monte Carlo (MCMC) approach was used to analyze region D or full-length VP1 gene sequences of GII/4 NoV populations isolated in Brazil or Japan, respectively. The results of these studies revealed that the expansion population growth model was the best to fit the data in both datasets. The dates of the most common recent ancestors revealed that these viruses can quickly emerge in a geographical location. A mean evolutionary rate of 1.21 x 10(-2) nucleotide substitution/site/year (s/s/y) was obtained for the VP1 gene using full-length sequences. This rate is higher than the rates reported for other rapidly evolving RNA. Roughly similar rates (1.44 x 10(-2)s/s/y) were found using region D sequences, revealing the suitability of this region for evolutionary studies, in agreement with previous reports. High evolutionary rates and fast population growth may have contributed to the vigorous initial transmission dynamics of the GII/4 NoV populations studied. PMID- 19559106 TI - Evaluating routine diagnostic imaging in acute appendicitis. AB - AIM: To evaluate the impact of selective imaging on clinical management of patients who present with symptoms suggesting acute appendicitis. MATERIALS AND METHODS: During a two-and-half year period, 941 consecutive patients with right lower quadrant pain were analyzed. Patients who underwent selective imaging were compared to those treated without further imaging. RESULTS: In 650 (69%) patients with right lower quadrant pain, diagnosis was based on medical history, physical and laboratory examination only. The diagnostic accuracy was 84%. Another 291 patients (31%) underwent selective imaging reaching a diagnostic accuracy of 71%. Ultrasound was conducted in 277 patients (sensitivity: 59%; specificity: 91%). CT scan was conducted in 43 patients (sensitivity: 100%; specificity: 95%). CONCLUSION: The present study shows that, in the majority of patients, appendicitis acuta can be diagnosed without the aid of imaging studies. In all these cases, high diagnostic accuracy rates and low morbidity rates were achieved. In all the other cases when clinical diagnosis is uncertain, further evaluation should include imaging. In our series ultrasound is of limited value; CT scan or diagnostic laparoscopy seems superior. PMID- 19559107 TI - Why farming with high tech methods should integrate elements of organic agriculture. AB - In the previous article [Ammann, K. (2008) Feature: integrated farming: why organic farmers should use transgenic crops. New Biotechnol. 25, 101-107], in a plea for the introduction of transgenic crops into organic and integrated farming, it was announced that the complementary topic, namely that high tech farmers should integrate elements of organic agriculture, will be a follow up. Some selected arguments for such a view are summarised here. Basically, they comprise a differentiated view on agro-biodiversity outside the field of production; landscape management methods to enhance biodiversity levels. Both elements are compatible with basic ideas of organic farming. First, Precision Farming is given as one example of the many ways to support agricultural production through high technology, with the aim of reducing energy input, maintaining excellent soil conditions and enhancing yield. It is clear from this analysis that modern agriculture and certain elements of organic-integrated agriculture are compatible. There are sectors of high tech farming, such as the introduction of a better recycling scheme and also a better focus on socio economic aspects, which need to be taken up seriously from organic-integrated farming, a system which puts a lot of emphasis on those elements and for which important research data are available. In the final part a new concept of dynamic sustainability is presented. PMID- 19559108 TI - Umbilical cord blood stem cells: towards a proteomic approach. AB - The first umbilical cord blood (UCB) transplant to a sibling with Fanconi's anaemia in 1988 represented a breakthrough in the field of transplantation. Thereon, several transplants have been performed with UCB-derived hematopoietic stem cells (HSCs) and a plethora of studies have investigated the plasticity of UCB-derived stem and progenitor cells. However, these studies have not been hitherto translated into clinical trials and, although UCB is routinely used as an alternative source of HSCs, no substantial advances have been made in the field of clinical regenerative medicine. The real deal is the lack of knowledge about the molecular processes governing the events of differentiation which transform immature UCB stem cells into terminally-committed hematopoietic, muscle, bone and nervous cells. In order to fill this void, several studies have been recently focused on the identification of the peculiar proteomic profile of UCB-derived stem cells. Hereby, we concisely review recent proteomic surveys addressing UCB-derived stem and progenitor cells. Notably, comparative studies detected a wider spectrum of proteins in immature cells rather than in more differentiated populations, as if maturation events could represent a bottleneck to protein expression. Future research projects should try to shed light on these processes and their completion could pave the way for unprecedented treatments. PMID- 19559109 TI - Improved antibiotic-free DNA vaccine vectors utilizing a novel RNA based plasmid selection system. AB - To ensure safety, regulatory agencies recommend elimination of antibiotic resistance markers from therapeutic and vaccine plasmid DNA vectors. Here, we describe the development and application of a novel antibiotic-free selection system. Vectors incorporate and express a 150 bp RNA-OUT antisense RNA. RNA-OUT represses expression of a chromosomally integrated constitutively expressed counter-selectable marker (sacB), allowing plasmid selection on sucrose. Sucrose selectable DNA vaccine vectors combine antibiotic-free selection with highly productive fermentation manufacturing (>1g/L plasmid DNA yields), while improving in vivo expression of encoded proteins and increasing immune responses to target antigens. These vectors are safer, more potent, alternatives for DNA therapy or vaccination. PMID- 19559110 TI - Novel Adenovirus type 5 vaccine platform induces cellular immunity against HIV-1 Gag, Pol, Nef despite the presence of Ad5 immunity. AB - Recombinant Adenovirus serotype 5 (Ad5) vectors have been used as vaccine platforms in numerous animal and human clinical studies. The immune response induced by Ad5 vaccines can be mitigated due to pre-existing Ad5 immunity. We previously reported the use of a novel Ad5 platform to induce cellular immune responses (CMI) against HIV-1 Gag in Ad5 hyper immune mice. Here, the effectiveness of the Ad5 [E1-, E2b-] vaccine platform was evaluated using a triad mixture of HIV-1 Gag, Pol, and Nef as antigenic transgenes. Broad CMI was induced following vaccination with the HIV-1 expressing vectors in Ad5 naive and Ad5 immunized mice. A mixture of the three vaccines induced CMI against each transgene product even in the presence of hyper Ad5 immunity. These studies revealed that CMI responses to immunization with Ad5 [E1-, E2b-]-gag, Ad5 [E1-, E2b-]-pol or Ad5 [E1-, E2b-]-nef vectors were transgene specific and did not induce CMI responses against irrelevant antigens such as carcinoembryonic antigen (CEA), herpes simplex virus glycoprotein B (HSV), cytomegalovirus (CMV) or influenza virus antigens. We are evaluating this recombinant triad viral vector as an HIV-1 vaccine in a non-human primate model and the data indicate that the vaccine is worthy of clinical evaluation. PMID- 19559111 TI - Immunogenicity of candidate chimeric DNA vaccine against tuberculosis and leishmaniasis. AB - Mycobacterium tuberculosis and Leishmania donovani are important intracellular pathogens, especially in Indian context. In India and other South East Asian countries, both these infections are highly endemic and in about 20% cases co infection of these pathogens is reported. For both these pathogens cell mediated immunity plays most important role. The available treatment of these infections is either prolonged or cumbersome or it is ineffective in controlling the outbreaks and spread. Therefore, potentiation of a common host defense mechanism can be used to prevent both the infections simultaneously. In this study we have developed a novel chimeric DNA vaccine candidate comprising the esat-6 gene of M. tuberculosis and kinesin motor domain gene of L. donovani. After developing this novel chimera, its immunogenicity was studied in mouse model. The immune response was compared with individual constructs of esat-6 and kinesin motor domain. The results showed that immunization with chimeric DNA vaccine construct resulted in stronger IFN-gamma and IL-2 response against kinesin (3012+/-102 and 367.5+/ 8.92pg/ml) and ESAT-6 (1334+/-46.5 and 245.1+/-7.72pg/ml) in comparison to the individual vaccine constructs. The reciprocal immune response (IFN-gamma and IL 2) against individual construct was lower (kinesin motor domain: 1788+/-36.48 and 341.8+/-9.801pg/ml and ESAT-6: 867.0+/-47.23 and 170.8+/-4.578pg/ml, respectively). The results also suggest that using the chimeric construct both proteins yielded a reciprocal adjuvant affect over each other as the IFN-gamma production against chimera vaccination is statistically significant (p<0.0001) than individual construct vaccination. From this pilot study we could envisage that the chimeric DNA vaccine construct may offer an attractive strategy in controlling co-infection of leishmaniasis and tuberculosis and have important implication in future vaccine design. PMID- 19559112 TI - Therapeutic vaccine IC41 as late add-on to standard treatment in patients with chronic hepatitis C. AB - We examined the effect of the hepatitis C virus (HCV) peptide vaccine IC41 on HCV specific T-cell responses and virological relapse rates in patients with chronic HCV genotype 1 infection when added to pegylated interferon plus ribavirin standard therapy. 35 patients received 6 vaccinations with IC41 from weeks 28 to 48 of standard antiviral treatment and were followed-up for another 6 months. IC41 vaccination did not prevent HCV-RNA relapse in patients with ongoing interferon standard treatment but HCV-specific T-cell responses were inducible and were associated with lower relapse rates. An increase of HCV-specific T-cell responses occurred in 73% of patients, responses were more frequent and stronger in patients with sustained virologic response than in patients who relapsed. Optimized vaccine responses may enhance sustained virologic response rates obtained with standard treatment of chronic hepatitis C. PMID- 19559113 TI - Use of MDCK cells for production of live attenuated influenza vaccine. AB - To develop a cell-based live attenuated influenza vaccine (LAIV) manufacturing process, several different cell lines were evaluated by comparing the titer of viruses after infection with LAIV strains. While several cell lines have been reported to support influenza virus replication, the degree of replication and the ability to support replication of LAIV strains have not been systematically examined. MDCK cells, which have been considered as potential substrates for influenza vaccine production were evaluated in addition to Vero, MRC-5, WI-38 and FRhL cells. MRC-5, WI-38 and FRhL cells produced low to moderate titers of virus with titers equal or below 5.0 log(10) TCID(50)/mL. Both Vero and MDCK cells could support a higher level of virus replication for certain strains, however, Vero cells only produced high titers when grown in the presence of serum. MDCK cells supported high levels of vaccine virus production for multiple different LAIV subtypes in both serum containing and serum-free media. These results suggest that MDCK cell-based production can be used as an alternative production platform to the currently used egg-based LAIV production system. PMID- 19559114 TI - Immunogenicity of an HPV-16 L2 DNA vaccine. AB - The ability to elicit cross-neutralizing antibodies makes human papillomavirus (HPV) L2 capsid protein a possible HPV vaccine. We examined and compared the humoral response of mice immunized with a HPV-16 L2 DNA vaccine or with HPV-16 L2 protein. The L2 DNA vaccine elicited a non-neutralizing antibody response unlike the L2 protein. L2 DNA vaccination suppressed the growth of L2-expressing C3 tumor cells, which is a T cell mediated effect, demonstrating that the lack of non-neutralizing antibody induction by L2 DNA was not caused by lack of T cell immunogenicity of the construct. PMID- 19559115 TI - Genotypification of bovine group A rotavirus in Mexico. AB - Bovine scours, frequently provoked by rotavirus infection, causes significant economic losses. Nevertheless, no information exists about the bovine rotavirus genotypes present in Mexico. This information is necessary for designing efficient vaccines. In this work, 128 samples from diarrheic calves were collected between 2005 and 2006 in 26 dairy and/or beef cattle herds located in 10 regions of Mexico, and analyzed for the presence of group A rotavirus. G and P genotypes were determined by PCR in rotavirus-positive samples (12/128). Three different genotype combinations were found, G10, P[11]; G6, P[5]; and G10, P[5]; in 67, 25 and 8% of the positive samples, respectively. Some rotavirus-positive animals had been vaccinated with an inactivated rotavirus strain of a different genotype. PMID- 19559117 TI - Emerging diseases, zoonoses and vaccines to control them. AB - Vaccination, when available, is undoubtedly the most cost-effective means of preventing and controlling, and even eradicating, infectious diseases. In recent years vaccination has also been used for other purposes in animal health, production and welfare, e.g. immunocastration. Vaccination of animals serves many different purposes, such as controlling animal infections and infestations, thus improving animal health and welfare; controlling anthropozoonoses and food poisoning in humans, thereby protecting public health; solving problems associated with antibiotic and anthelmintic resistance; helping to leave food producing animals free of chemical residues; protecting the environment and biodiversity and ensuring animal farming sustainability. The problem is nevertheless more complex when facing emerging or re-emerging infections particularly zoonotic ones. PMID- 19559116 TI - Rational design and immunogenicity of liposome-based diepitope constructs: application to synthetic oligosaccharides mimicking the Shigella flexneri 2a O antigen. AB - We have designed chemically defined diepitope constructs consisting of liposomes displaying at their surface synthetic oligosaccharides mimicking the O-antigen of the Shigella flexneri 2a lipopolysaccharide (B-cell epitope) and influenza hemagglutinin peptide HA 307-319 (Th epitope). Using well controlled and high yielding covalent bioconjugation reactions, the two structurally independent epitopes were coupled to the lipopeptide Pam(3)CAG, i.e. a TLR2 ligand known for its adjuvant properties, anchored in preformed vesicles. The synthetic construct containing a pentadecasaccharide corresponding to three O-antigen repeating units triggered T-dependent anti-oligosaccharide and anti-S. flexneri 2a LPS antibody responses when administered i.m. to BALB/c mice. Moreover, the long-lasting anti LPS antibody response afforded protection against a S. flexneri 2a challenge. These results show that liposome diepitope constructs could be attractive alternatives in the development of synthetic carbohydrate-based vaccines. PMID- 19559118 TI - Influenza control in the 21st century: Optimizing protection of older adults. AB - Older adults (> or =65 years of age) are particularly vulnerable to influenza illness. This is due to a waning immune system that reduces their ability to respond to infection, which leads to more severe cases of disease. The majority ( approximately 90%) of influenza-related deaths occur in older adults and, in addition, catastrophic disability resulting from influenza-related hospitalization represents a significant burden in this vulnerable population. Current influenza vaccines provide benefits for older adults against influenza; however, vaccine effectiveness is lower than in younger adults. In addition, antigenic drift is also a concern, as it can impact on vaccine effectiveness due to a mismatch between the vaccine virus strain and the circulating virus strain. As such, vaccines that offer higher and broader protection against both homologous and heterologous virus strains are desirable. Approaches currently available in some countries to meet this medical need in older adults may include the use of adjuvanted vaccines. Future strategies under evaluation include the use of high-dose vaccines; novel or enhanced adjuvantation of current vaccines; use of live attenuated vaccines in combination with current vaccines; DNA vaccines; recombinant vaccines; as well as the use of different modes of delivery and alternative antigens. However, to truly evaluate the benefits that these solutions offer, further efficacy and effectiveness studies, and better correlates of protection, including a precise measurement of the T cell responses that are markers for protection, are needed. While it is clear that vaccines with greater immunogenicity are required for older adults, and that adjuvanted vaccines may offer a short-term solution, further research is required to exploit the many other new technologies. PMID- 19559120 TI - Production of yellow fever virus in microcarrier-based Vero cell cultures. AB - In this work, the propagation of the 17DD yellow fever virus in Vero cells grown on Cytodex-1 microcarriers was evaluated. After verifying that upon infection the virus adsorption step could be performed under continuous agitation, experiments were carried out in spinners and sparged lab-scale stirred-tank bioreactor to evaluate the use of a commercial serum-free medium (VP-SFM) and to investigate the effects of multiplicity of infection (MOI) and time of infection (TOI) on virus production. Virus titers as high as 8.4 x 10(8)pfu/mL were obtained upon infection with MOI of 0.02 and TOI of 3 days, using the serum-free medium in the sparged bioreactor. PMID- 19559119 TI - In silico-accelerated identification of conserved and immunogenic variola/vaccinia T-cell epitopes. AB - Epitopes shared by the vaccinia and variola viruses underlie the protective effect of vaccinia immunization against variola infection. We set out to identify a subset of cross-reactive epitopes using bioinformatics and immunological methods. Putative T-cell epitopes were computationally predicted from highly conserved open reading frames from seven complete vaccinia and variola genomes using EpiMatrix. Over 100 epitopes bearing low human sequence homology were selected and assessed in HLA binding assays and in T-cell antigenicity assays using PBMCs isolated from Dryvax-immunized subjects. This experimental validation of computational predictions illustrates the potential for immunoinformatics methods to identify candidate immunogens for a new, safer smallpox vaccine. PMID- 19559121 TI - Respiratory syncytial virus subunit vaccine based on a recombinant fusion protein expressed transiently in mammalian cells. AB - Although respiratory syncytial virus (RSV) causes severe lower respiratory tract infection in infants and adults at risk, no RSV vaccine is currently available. In this report, efforts toward the generation of an RSV subunit vaccine using recombinant RSV fusion protein (rRSV-F) are described. The recombinant protein was produced by transient gene expression (TGE) in suspension-adapted human embryonic kidney cells (HEK-293E) in 4 L orbitally shaken bioreactors. It was then purified and formulated in immunostimulating reconstituted influenza virosomes (IRIVs). The candidate vaccine induced anti-RSV-F neutralizing antibodies in mice, and challenge studies in cotton rats are ongoing. If successful in preclinical and clinical trials, this will be the first recombinant subunit vaccine produced by large-scale TGE in mammalian cells. PMID- 19559123 TI - Suspension-Vero cell cultures as a platform for viral vaccine production. AB - Since Vero cells are currently considered as an acceptable cell substrate to produce a wide range of viruses, we developed a virus production platform using Vero cells adapted to grow in suspension in serum-free media. After adapting anchorage-dependent Vero cells to grow as a free-cell suspension, vesicular stomatitis virus, herpes simplex virus 1 and polio virus 1 production rates were evaluated in batch cultures using spinner flasks and perfused cultures in a bioreactor. The achieved results constitute valuable information for the development of a low-cost high-productivity process using a suspension culture of Vero cells to produce viral vaccines. PMID- 19559124 TI - Carbohydrate: more than complex. PMID- 19559122 TI - Adjuvant activities of saponins from traditional Chinese medicinal herbs. AB - New generation vaccines such as recombinant, antigen purified and DNA vaccines are poorly immunogenic due to the lack of an innate immune stimulus. Therefore, search of new adjuvants for these vaccines has become a topic of interesting. In new adjuvant development, saponins are outstanding candidates. Recently, increased attention has been received on plant-derived saponins in search of new adjuvant candidates from traditional Chinese medicinal herbs such as Panax ginseng, Astragalus species, Panax notoginseng,Cochinchina momordica, Glycyrrhiza uralensis and Achyranthes bidentata. Many of the saponins have been found to have adjuvant effects on purified protein antigens. The chemical structures of the saponins are related to their adjuvant activities, and influence the nature of the immune responses. Saponin adjuvants have been reported to stimulate secretion of a broad range of cytokines, suggesting that saponins may act by triggering innate immunity. As these plant-originated adjuvants may promote different branches of the immune system, they have the potential to be used in design of new vaccines so as to induce a desired immune response. PMID- 19559125 TI - To eat or not to eat? Food safety in the United States. PMID- 19559127 TI - Taking trans fat off the menu: what you can learn from trans-fat bans at Sheikh Khalifa Medical City and the Cleveland Clinic. PMID- 19559129 TI - ADA and health reform. PMID- 19559128 TI - Make the most of your professional development portfolio. PMID- 19559130 TI - The diabetes epidemic: what can we do? PMID- 19559131 TI - Carbohydrate intake and obesity: an association that needs "refining". PMID- 19559133 TI - Trends in nutrient intake among adults with diabetes in the United States: 1988 2004. AB - BACKGROUND: Weight loss through dietary modification is key to type 2 diabetes self-management, yet few nationally representative data exist on dietary trends among people with diabetes. OBJECTIVE: To examine dietary changes, via nutrient intakes, among US adults with diabetes between 1988 and 2004. DESIGN: Nutrition data from the cross-sectional National Health and Nutrition Examination Surveys (Phase I: 1988-1990 and Phase II: 1991-1994) and 1999-2004 of adults with self reported diabetes were examined. Twenty-four-hour dietary recall data were used to assess changes in energy; carbohydrate; protein; total, saturated, polyunsaturated, and monounsaturated fat; cholesterol; fiber; sodium; and alcohol intake. STATISTICAL ANALYSES: Consumption of total energy and specific nutrients per day were estimated by survey, controlled for age and sex, using multiple linear regression and adjusted means (with standard errors). RESULTS: Between 1988 and 2004 there was no significant change in self-reported total energy consumption among adults with self-reported diabetes (1,941 kcal/day in 1988-1990 to 2,109 kcal/day in 2003-2004, P for trend=0.22). However, there was a significant increase in the consumption of carbohydrate (209 g/day in 1988-1990 to 241 g/day in 2003-2004; P for trend=0.02). In analyses stratified by age group, changes in dietary consumption were noted among persons aged 45 to 64 years; specifically, increases in total energy (1,770 to 2,100 kcal/day, P for trend =0.01) and carbohydrate consumption (195 to 234 g/day, P for trend=0.02). CONCLUSIONS: Despite recommendations to lose weight, daily energy consumption by individuals with diabetes showed no significant change, except in individuals aged 45 to 64 years, where an increase was observed. Overall, there was an increase in carbohydrate consumption. Emphasizing the equal importance of energy reduction and changes in dietary composition for people with diabetes is important for optimal self-management. PMID- 19559132 TI - Carbohydrate intake and overweight and obesity among healthy adults. AB - BACKGROUND: Little is known about the dietary habits of people with optimal body weight in communities with high overweight and obesity prevalence. OBJECTIVE: To evaluate carbohydrate intake in relation to overweight and obesity in healthy, free-living adults. DESIGN: We used a cross-sectional analysis. SUBJECTS/SETTING: The Canadian Community Health Survey Cycle 2.2 is a cross-sectional survey of Canadians conducted in 2004-2005. There were 4,451 participants aged 18 years and older with anthropometric and dietary data and no comorbid conditions in this analysis. MAIN OUTCOME MEASURES: Outcome variables were body mass index (BMI; calculated as kg/m(2)) and overweight or obesity status (dichotomous) defined as BMI > or =25 compared with BMI <25 based on measured height and weight. Diet was evaluated by 24-hour dietary recall based on the Automated Multi-Pass Method. STATISTICAL ANALYSES PERFORMED: Weighted regression models with bootstrapping and cubic splines were used. Outcome variables were BMI and overweight or obesity, and predictors were daily nutrient intake. Adjustment for total energy intake, age, leisure time energy expenditure, sex, smoking, education, and income adequacy was performed. RESULTS: Risk of overweight and obesity was decreased in all quartiles of carbohydrate intake compared to the lowest intake category (multivariate odds ratio quartile 2=0.63; 95% confidence interval: 0.49 to 0.90; odds ratio quartile 3=0.58; 95% confidence interval: 0.41 to 0.82; odds ratio quartile 4=0.60; 95% confidence interval: 0.42 to 0.85). Spline analyses revealed lowest risk among those consuming 290 to 310 g/day carbohydrates. CONCLUSIONS: Consuming a low-carbohydrate (approximately <47% energy) diet is associated with greater likelihood of being overweight or obese among healthy, free-living adults. Lowest risk may be obtained by consuming 47% to 64% energy from carbohydrates. PMID- 19559134 TI - N-nitroso compounds: assessing agreement between food frequency questionnaires and 7-day food records. AB - BACKGROUND: N-nitroso compounds are recognized as important dietary carcinogens. Accurate assessment of N-nitroso intake is fundamental to advancing research regarding its role with cancer. Previous studies have not used a quantitative database to estimate the intake of these compounds in a US population. OBJECTIVE: To address this gap, a database of N-nitroso values was developed in conjunction with an existing food frequency questionnaire (FFQ). In this article we report on the relative validity of the FFQ instrument modified to estimate intake of N nitroso compounds. DESIGN: Intake estimates of 23 N-nitroso compounds from the FFQ were compared with those from 7-day food records in a cross-sectional study conducted from January 2005 through June 2006. SUBJECTS: A sample of 98 healthy adult subjects (aged 50.42+/-12.84 years) completed an FFQ and then recorded foods and beverages consumed on 7-day food records. RESULTS: Crude and energy adjusted N-nitroso compounds intakes were significantly higher in the FFQ than the 7-day food records (P<0.001). Spearman correlations for crude and energy adjusted N-nitroso intakes ranged from 0.004 to 0.48. By tertiles of N-nitiroso compounds, there was moderate agreement (kappa>0.30) for five compounds. Higher estimates of N-nitroso compounds by FFQ was explained by a greater proportion of subjects who reported eating foods high in N-nitroso compounds on FFQ than reported on 7-day food records. CONCLUSION: The modified FFQ with N-nitroso values is a useful tool for assessing N-nitroso intakes relative to a group, and captures all food items with N-nitroso compounds, including those foods with high concentrations and eaten sporadically. PMID- 19559135 TI - Validity and calibration of food frequency questionnaires used with African American adults in the Jackson Heart Study. AB - OBJECTIVE: To examine the relative validity of two food frequency questionnaires (FFQs) developed for use in investigating diet and disease relationships within the adult African-American population in the southern United States. DESIGN: Cross-sectional analyses of dietary nutrient intake data, comparing four 24-hour dietary recalls with an FFQ developed by the Lower Mississippi Delta Nutrition Intervention Research Initiative, and its shorter version adapted for use in the Jackson Heart Study. SUBJECTS: A representative subset of participants (n=499, aged 35 to 81 years) from the baseline Jackson Heart Study cohort (N=5,302) was selected for this study. Data collection took place between winter 2000 and spring 2004. STATISTICAL ANALYSES: Pearson's correlation coefficients (energy adjusted and de-attenuated) for 26 nutrients estimates from each of the FFQs, comparing them with the mean of four 24-hour dietary recalls. The ability of the FFQs to rank individuals based on nutrient intakes was compared to that of the mean of four 24-hour dietary recalls and attenuation coefficients were also calculated. RESULTS: Median nutrient intake estimates tended to be higher on the long and lower on the short FFQ compared to the median for the mean of four 24 hour dietary recalls. Energy adjusted and deattenuated correlations of FFQ intake estimates with recalls ranged from 0.20 for sodium to 0.70 for carbohydrate for the short FFQ and from 0.23 for polyunsaturated fat to 0.75 for dietary fiber and magnesium for the long. Attenuation coefficients for men on average were 0.42 for the short and 0.49 for the long FFQ. For women, these were 0.31 for the short and 0.42 for the long FFQ. CONCLUSIONS: Both FFQs appear to be reasonably valid for assessment of dietary intake of adult African Americans in the South. The Lower Mississippi Delta Nutrition Intervention Research Initiative FFQ exhibited higher intake estimates and stronger correlations with recalls than the Jackson Heart Study FFQ for most nutrients analyzed, more so for women than men. PMID- 19559137 TI - The role of diet in symptoms of irritable bowel syndrome in adults: a narrative review. AB - This review summarizes what is known about the effect of diet on irritable bowel syndrome (IBS) symptoms emphasizing data from randomized, controlled clinical trials. Studies suggest that IBS symptoms in one quarter of patients may be caused or exacerbated by one or more dietary components. Recent studies indicate that a diet restricted in fermentable, poorly absorbed carbohydrates, including fructose, fructans (present in wheat and onions), sorbitol, and other sugar alcohols is beneficial, but confirmatory studies are needed. Despite a long history of enthusiastic use, fiber is marginally beneficial. Insoluble fiber may worsen symptoms. Some patients with IBS, especially those with constipation, will improve with increased intake of soluble fiber. Prebiotic fibers have not been adequately tested. Daily use of peppermint oil is effective in relieving IBS symptoms. The usefulness of probiotics in the form of foods such as live-culture yogurt and buttermilk for IBS symptoms is not established. In clinical practice, it is very difficult to establish that a patient's symptoms result from an adverse reaction to food. A double blind placebo-controlled food challenge is the most reliable method, but it is not suitable for routine clinical use. A modified exclusion diet and stepwise reintroduction of foods or trials of eliminating classes of food may be useful. PMID- 19559136 TI - Differences between food group reports of low-energy reporters and non-low-energy reporters on a food frequency questionnaire. AB - BACKGROUND: Low-energy reporters (LERs) and non-LERs differ with respect to several characteristics, including self-reported intake of foods. Limited data exist regarding food intake difference between LERs and non-LERs identified using doubly labeled water (DLW). OBJECTIVE: In the Observing Protein and Energy Nutrition Study (September 1999-March 2000), differences were examined between food group reports of LERs and non-LERs on a food frequency questionnaire (FFQ) (n=440). DESIGN: LERs were identified using DLW. Responses of LERs (n=220) and non-LERs (n=220) for 43 food groups on the FFQ were examined in three ways: whether they reported consuming a food group (yes/no), how frequently they reported consuming it (times per day), and the reported portion size (small, medium, or large). Analyses were adjusted for total energy expenditure from DLW. RESULTS: LERs, compared to non-LERs, were less likely to report consumption for one food group among women (soft drinks/regular). Among men, there was no difference between LERs and non-LERs with respect to reporting consumption of food groups. Reported mean daily frequency of consumption was lower among LERs compared with non-LERs for 23 food groups among women and 24 food groups among men (18 food groups were similar in men and women). In addition, reported mean portion sizes were smaller for LERs compared with non-LERs for six food groups among women and five food groups among men (three food groups were similar in men and women). Results varied minimally by sex and body mass index. CONCLUSIONS: LERs, compared with non-LERs, were more likely to differ regarding their reported frequency of consumption of food groups than their reported consumption (yes/no) or portion size of food groups. Results did not vary greatly by sex or body mass index. It still remains unclear whether improvement in questionnaire design or additional tools or methods would lead to a decrease in differential reporting due to LER status on an FFQ. PMID- 19559138 TI - Food use in middle and high school fundraising: does policy support healthful practice? Results from a survey of Minnesota school principals. AB - This descriptive, cross-sectional study aimed to examine classroom, school-wide, and club/sports teams fundraising policies and practices of middle and high schools; concordance between policy and practice; and associations between healthful policy/practice scores and selected school characteristics. In 2006, principals/designees of middle (n=45) and high (n=71) schools in the St Paul/Minneapolis, MN, metropolitan area completed a self-administered mailed survey. Schools were attended by a convenience sample of students (n=349) participating in a longitudinal measurement study of children and their environments to assess obesity-related factors. Descriptive statistics, chi(2) tests, and multivariate linear regression were used to examine variables and associations of interest. Across schools, 50% had policies addressing the nutrient quality of food and drink items used in fundraising or disallowed food use for fundraising. About one third used chocolate, candy, and high-fat baked goods for classroom and school-wide fundraising; 60% sold these items for club/sports teams fundraising. More middle than high schools reported healthful fundraising policies or practices, as well as greater concordance between policies and practices. For all fundraising activities, high schools had significantly lower healthful policy/practice scores than middle schools (P<0.01). For school-wide fundraising, scores were significantly lower for public than private schools (P=0.02). Policies to regulate food used for fundraising were common and most supported healthful practice, particularly in middle schools. However, use of foods high in fat and added sugars remains a prevalent fundraising practice, especially in high schools and for club/sports teams, and requires additional attention. PMID- 19559139 TI - A garden pilot project enhances fruit and vegetable consumption among children. AB - Fruit and vegetable intake among children is inadequate. Garden-based nutrition education programs may offer a strategy for increasing fruit and vegetable intake in children. A 12-week pilot intervention was designed to promote fruit and vegetable intake among 4th to 6th grade children (n=93) attending a YMCA summer camp. Children participated in garden-based activities twice per week. Weekly educational activities included fruit and vegetable taste tests, preparation of fruit and vegetable snacks, and family newsletters sent home to parents. The pilot intervention was evaluated using a pre and post survey to determine participant satisfaction and the short-term impacts of the program. The process evaluation focused on program satisfaction, whereas the short-term impact evaluation assessed fruit and vegetable exposure, preference, self-efficacy, asking behavior, and availability of fruits and vegetables in the home. Data from the impact evaluation were compared from baseline to follow-up using McNemar's test (dichotomous variables) and Wilcoxon signed rank test (scales/continuous variables). Children reported high levels of enjoyment in the intervention activities. Most children (97.8%) enjoyed taste-testing fruits and vegetables. Children also liked preparing fruit and vegetable snacks (93.4%), working in their garden (95.6%), and learning about fruits and vegetables (91.3%). Impact data suggest that the intervention led to an increase in the number of fruits and vegetables ever eaten (P<0.001), vegetable preferences (P<0.001), and fruit and vegetable asking behavior at home (P<0.002). Garden-based nutrition education programs can increase fruit and vegetable exposure and improve predictors of fruit and vegetable intake through experiential learning activities. Participation in the "seed to table" experience of eating may help promote healthful eating behaviors among youth. Food and nutrition professionals should consider garden-based nutrition education programs that connect children with healthful foods through fun, hands-on activities. PMID- 19559140 TI - A Fresh Fruit and Vegetable Program improves high school students' consumption of fresh produce. AB - Low fruit and vegetable intake may be associated with overweight. The United States Department of Agriculture implemented the Fresh Fruit and Vegetable Program in 2006-2007. One Houston-area high school was selected and received funding to provide baskets of fresh fruits and vegetables daily for each classroom during this period. This study assessed the impact of the program on students' fruit and vegetable intake. At program end (May 2007), fruit and vegetable intake surveys were distributed to students at the intervention school as well as at a comparison high school that did not receive the program. Surveys, which were completed anonymously, were received from 34% of intervention and 42% of comparison school students. The students were classified as to whether they met the recommended daily intake of fruit, 100% fruit juice, and vegetables. The probability of meeting the recommendations was assessed with logistic regression analyses, controlling for sex, age, and ethnicity. Compared with the comparison control school students, intervention school students were more likely to report eating fruit and drinking 100% fruit juice at least two times per day (39.3% vs 27.3%; P<0.05) and consuming total fruit, juice, and vegetables (22% vs 18.4%; P<0.05) five or more times per day in the preceding 7 days. More intervention school students (59.1%) than comparison school students (40.9%) reported eating fruit at least one time per day (P<0.05). There were no group differences in vegetable intake. Fresh fruit and vegetable distribution programs provide the opportunity for students to taste a variety of fruits and vegetables, and may improve consumption of these foods by adolescents. PMID- 19559141 TI - Development of a hand-held personal digital assistant-based food diary with food photographs for Japanese subjects. AB - Hand-held personal digital assistant (PDA)-based food diaries have been developed for self-monitoring of dietary intake, but the accuracy of these diaries is unclear for patients with diabetes. The aim of the study was to assess the accuracy and feasibility of use of a new PDA-based food diary, including food photographs. The study included 44 Japanese participants without diabetes (mean age 23 years) and 16 Japanese patients with type 2 diabetes (mean age 53 years). The PDA-based food diary was used for 7 consecutive days. Information about all dietary intake on the 7th day of PDA self-monitoring was collected by a 24-hour recall interview on the 8th day. The PDA-based data for dietary intake on the 7th day were then compared to the 24-hour recall data for the same period. Feasibility was assessed based on the frequency and timeliness of self monitoring. There was no significant difference in daily totals for energy, protein, carbohydrate, and fat between the two methods in each group. Pearson's correlation and intraclass correlation coefficients showed strong significant relationships for all variables between the two methods in both groups. Bland Altman plots did not indicate any bias in estimated daily caloric intake. Participants recorded 98% of their meals in the PDA, with 75% of entries recorded within 6 hours after the meal starting time. The findings suggest that the PDA based food diary is a potential clinical method to estimate dietary intake and may be a beneficial tool for self-monitoring of dietary intake. PMID- 19559142 TI - Validation of two food frequency questionnaires for dietary calcium assessment. AB - Easily utilized questionnaires estimating dietary calcium intake would be a valuable asset to promoting skeletal health as well as a helpful research tool. Two calcium questionnaires, one online and one printed, were each compared with dietary calcium intake measured by a 3-day diet record. Women completed the questionnaires in a randomized order and kept a 3-day food record at home, returning it by mail. The ethnicity of the 140 female study participants was 102 white, 12 African American, 16 Asian, and 10 Latina. The mean age (+/-standard deviation) was 49+/-15 years, with a range of 22.7 to 89.9 years. Measures used included an online calcium quiz, a printed calcium food frequency questionnaire, and one 3-day diet record. Data analysis was done using Pearson correlation coefficients. Sub-analyses were conducted by ethnicity. The correlation between each questionnaire and the daily calcium intake from the diet records was 0.37, P<0.001. Among the ethnic subgroups, the correlations were highest for African American women, followed by white women. There was also a trend toward higher correlations in younger women. These simple calcium assessment tools, taking fewer than 5 minutes to complete, have correlation values with diet records similar to more complex food frequency questionnaires reported in the literature. These questionnaires may provide valuable tools to estimate calcium intake in research, clinical, and community settings. PMID- 19559143 TI - Monitoring energy intake: a hand-held personal digital assistant provides accuracy comparable to written records. AB - New approaches to assess energy intake (EI) may have advantages over traditional written methods, but validity of these emerging methodologies must be demonstrated. This exploratory study compared EI obtained using a hand-held personal digital assistant (PDA) and traditional written records with total energy expenditure measured by doubly labeled water (TEE(DLW)). Twenty-six volunteers (aged 23+/-4 years, body mass index [calculated as kg/m(2)] 24+/-2) participated in a randomized (either PDA or written record group) and matched (for sex, age, and body mass index) study for 7 consecutive days between June 2005 and April 2006 to record EI. Group comparisons were made with t and Mann Whitney U tests. Bland-Altman plots were used to compare limits of agreement between methods. Volunteers remained weight stable during the study period (0.2+/ 0.8 kg; P>0.05). Reported EI by written record and PDA were similar to TEE(DLW); 105% vs 92% of TEE(DLW), respectively (P>0.05). There was a significant relationship between reported EI by PDA and TEE(DLW) (r=0.60, P<0.05), but not for written record (r=0.45, P>0.05). Limits of agreement indicated both written record and PDA had large variability (range 1,394 to -1,472 kcal/day). Findings suggest the bias in using a PDA is similar to that observed when using a written record for estimation of EI in weight-stable volunteers. PMID- 19559144 TI - Association of depressive symptoms and lifestyle behaviors among Latinos at risk of type 2 diabetes. AB - Little is known about depression among Latinos at risk for type 2 diabetes. The purpose of this cross-sectional study was to determine the rate of depression in Latinos at risk for type 2 diabetes and to examine the associations between depressive symptoms, diet, physical activity, and body mass index (BMI; calculated as kg/m(2)). Latinos at risk for type 2 diabetes (n=210) were surveyed from July 2007 to August 2008. Depressive symptoms were assessed using the Center for Epidemiological Studies Depression Scale (CES-D). Three 24-hour recalls were used to assess diet and physical activity. Linear regression analyses were used to examine the association between depression, BMI, and lifestyle factors. The sample (78% female) was largely of Caribbean origin (83%) and mean age was 52 years (standard deviation [SD]=11 years). Mean BMI was 34.2 (SD=5.9) and 77% were obese. Average CES-D score was 16.3 (SD=11.3; range=0-45) and 50% had CES-D score > or =16, suggesting clinical depression. Higher CES-D scores were associated with lower diet quality (P<0.05), but were not associated with BMI or physical activity. Depression and lower dietary quality can contribute to risk for type 2 diabetes among Latinos. PMID- 19559145 TI - Validation of a semi-quantitative food frequency questionnaire measuring dietary vitamin K intake in elderly people. AB - The study objective was to validate a semi-quantitative food frequency questionnaire (FFQ) specifically designed to measure dietary vitamin K intake. A 50-item FFQ was interviewer-administered and compared with data previously obtained from 5-day food records. Thirty-nine community-dwelling healthy men and women aged 65 to 85 years were recruited from the Montreal metropolitan area. Absolute and relative agreements between methods were assessed. Vitamin K intake measured by the vitamin K FFQ (mean+/-standard deviation; 222+/-186 microg/day) was significantly higher than that obtained by food records (135+/-153 microg/day; P<0.001). Bland-Altman analysis on log(10)-transformed data indicated that vitamin K intake from vitamin K FFQ was 2.26 times (95% confidence interval: 1.90 to 2.67) higher than food records, limits of agreement ranging from 0.80 to 6.35. However, correlation between methods was strong and highly significant (r=0.83; P<0.001). Cross-classification also showed that 72% of participants were correctly classified into thirds and only 8% were grossly miscategorized. Weighted kappa value (kappa=0.60) also indicated a good relative agreement. In light of these results, the vitamin K FFQ is a valid tool for ranking individuals according to their vitamin K intake. The poor absolute agreement likely results from the inability for food records to adequately measure the usual intake of episodically consumed foods, particularly those high in vitamin K. The vitamin K FFQ will be useful in large-scale, population-based research on vitamin K and disease as well as in clinical practice, especially that focusing on anticoagulant therapy. PMID- 19559146 TI - A comprehensive coding system to measure the quality of school wellness policies. AB - In 2006, all local education agencies in the United States participating in the National School Lunch Program were required to establish school wellness policies that covered nutrition education, nutrition standards for school foods, and physical activity. The purpose of this psychometric study was to develop and evaluate the properties of a comprehensive and quantitative coding system to evaluate the quality of these policies. A 96-item coding tool was developed to evaluate seven goal areas: nutrition education, standards for US Department of Agriculture child nutrition programs and school meals, nutrition standards for competitive and other foods and beverages, physical education, physical activity, communication and promotion, and evaluation. Each goal area subscale and the total scale were scored on two dimensions: comprehensiveness and strength. Reliability was assessed by having pairs of researchers from four different states code a sample of 60 polices between July 2007 and July 2008. Goal area subscales were internally reliable (Cronbach's alpha=.60 to .93). Adequate interrater reliability scores were obtained at each level of scoring: total comprehensiveness and strength scores (intraclass correlation coefficient 0.82), subscale scores (intraclass correlation coefficient 0.70), and individual items (intraclass correlation coefficient 0.72). This coding system provided a reliable method for analyzing and comparing school district wellness policies in single or multistate studies. PMID- 19559148 TI - What are the primary nutritional issues for a patient with Parkinson's disease? PMID- 19559147 TI - Development of symptomatic cardiovascular disease after self-reported adherence to the Atkins diet. PMID- 19559149 TI - Cardiac pheochromocytoma presenting as shortness of breath. PMID- 19559151 TI - Appropriate antibiotics of Weil disease. PMID- 19559152 TI - Gastrointestinal bleeding: one complaint, two culprits. PMID- 19559153 TI - Atypical pyoderma gangrenosum presenting as paronychia. PMID- 19559154 TI - More study needed on morning erections and erectile dysfunction. PMID- 19559155 TI - Research productivity of MD-PhDs. PMID- 19559156 TI - It's time to bail out seniors trapped in the Medicare donut hole! PMID- 19559157 TI - "Hey, doc, is it OK for me to drink coffee?". PMID- 19559158 TI - Osteoporosis in inflammatory bowel disease. AB - Osteoporosis commonly afflicts patients with inflammatory bowel disease, and many factors link the 2 states together. A literature review was conducted about the pathophysiology of osteoporosis in relation to inflammatory bowel disease. Screening guidelines for osteoporosis in general as well as those directed at patients with inflammatory bowel disease are reviewed, as are currently available treatment options. The purpose of this article is to increase physician awareness about osteopenia and osteoporosis occurring in patients with inflammatory bowel disease and to provide basic, clinically relevant information about the pathophysiology and guidelines to help them treat these patients in a cost effective manner. PMID- 19559161 TI - Erythema multiforme syndrome. PMID- 19559160 TI - Pharmacological management of psychosis in elderly patients with parkinsonism. AB - Parkinsonism is a characteristic feature of Parkinson's disease and dementia with Lewy bodies and is commonly seen in Alzheimer's disease. Psychosis commonly appears during the course of these illnesses. Treatment of parkinsonism with antiparkinsonian medications constitutes an additional risk factor for the appearance or worsening of psychosis. Conversely, treatment of psychosis with antipsychotic drugs in patients with parkinsonism might worsen the underlying movement disorder, especially in the elderly. In this article, we review parkinsonian conditions in the elderly and offer guidelines to assess and manage comorbid psychosis. We focus on the pharmacologic management of psychosis with atypical antipsychotic medications and briefly review the role of acetylcholinesterase inhibitors. PMID- 19559159 TI - Inflammation, coagulation, and the pathway to frailty. AB - There are inevitable physiologic changes associated with advancing age, yet for some people these changes are exaggerated, and as a result a phenotype emerges recognized as "frailty." Why some people become frail and others do not remains incompletely understood. Although chronic illnesses are common among frail elderly persons, some will develop all of the phenotypic features without a diagnosed underlying disease. It has been recognized that certain proinflammatory cytokines and coagulation factors are elevated to a greater extent in those who are frail than in age-matched nonfrail individuals. In this review, we provide an overview of current research in the biology of frailty with particular emphasis on the role of inflammatory pathways and disordered coagulation in its pathogenesis. PMID- 19559162 TI - Hole in one. PMID- 19559163 TI - Transplant troubles. PMID- 19559165 TI - An unusual case of syncope. PMID- 19559164 TI - Bugs and blood cells. PMID- 19559167 TI - Effect of statin adherence on cerebrovascular disease in primary prevention. AB - BACKGROUND: Evidence from meta-analyses shows that statin therapy reduces all cause mortality and nonhemorrhagic strokes. Nonadherence to statins may reduce this protective effect. The association between statin adherence and incidence of cerebrovascular disease remains unexplored outside the context of clinical trials. OBJECTIVE: To evaluate the impact of statin adherence on the occurrence of cerebrovascular disease in a real clinical setting. METHODS: A cohort of 112,092 patients was reconstructed using the Regie d'assurance maladie du Quebec and Med-Echo databases. The Regie d'assurance maladie du Quebec database contains information from 3 types of health-related data, such as demographic information, medical data, and the prescription claims file. The Med-Echo database contains data on acute care hospitalizations on all Quebec residents. All patients without cardiovascular disease aged 45-85 years who were newly treated with statins between 1999 and 2004 were eligible. A nested case-control design was used to study the occurrence of cerebrovascular disease. Adherence level was reported as a medication possession ratio. Conditional logistic regression models were used to estimate the rate ratio of cerebrovascular disease, adjusting for different covariables. RESULTS: The mean patient age was 63 years; 49% had hypertension, 21% had diabetes, and 41% were males. Nonadherence was prevalent because only 55% of the patients were exposed to > or = 80% of the medication during follow-up. We did not observe any major differences, defined as more than 5%, between the groups, except for the sex, diabetes, and hypertension. High level of adherence to statins was associated with a reduction of cerebrovascular events (rate ratio: 0.74; 95% confidence interval, 0.65-0.84). CONCLUSIONS: Our study suggests a relatively low level of adherence to statins, but more importantly, that adherence is associated with a risk reduction for cerebrovascular disease. Adherence to statin therapy needs to be improved, so that patients can benefit from the full protective effects of statin therapy. PMID- 19559166 TI - The effect of a hypertension self-management intervention on diabetes and cholesterol control. AB - BACKGROUND: Most patient chronic disease self-management interventions target single-disease outcomes. We evaluated the effect of a tailored hypertension self management intervention on the unintended targets of glycosylated hemoglobin (HbA1c) and low-density lipoprotein cholesterol (LDL-C). METHODS: We evaluated patients from the Veterans Study to Improve the Control of Hypertension, a 2-year randomized controlled trial. Patients received either a hypertension self management intervention delivered by a nurse over the telephone or usual care. Although the study focused on hypertension self-management, we compared changes in HbA1c among a subgroup of 216 patients with diabetes and LDL-C among 528 patients with measurements during the study period. Changes in these laboratory values over time were compared between the 2 treatment groups using linear mixed effects models. RESULTS: For the patients with diabetes, the hypertension self management intervention resulted in a 0.46% reduction in HbA1c over 2 years compared with usual care (95% confidence interval, 0.04%-0.89%; P = .03). For LDL C, there was a minimal 0.9 mg/dL between-group difference that was not statistically significant (95% confidence interval, -7.3-5.6 mg/dL; P = .79). CONCLUSIONS: There was a significant effect of the self-management intervention on the unintended target of HbA1c, but not LDL-C. Chronic disease self-management interventions might have "spill-over" effects on patients' comorbid chronic conditions. PMID- 19559168 TI - Breastfeeding in infancy and adult cardiovascular disease risk factors. AB - BACKGROUND: Public health recommendations advocate breastfeeding in infancy as a means to reduce obesity in later life. Several prior studies relating breastfeeding to cardiovascular risk factors have been limited by lack of adjustment for maternal and participant confounding factors. METHODS: We ascertained breastfeeding history via questionnaire from mothers enrolled in the Framingham Offspring Study. In their young to middle-aged adult children enrolled in the Framingham Third Generation, we examined the relations between maternal breastfeeding history (yes, no) and cardiovascular risk factors, including body mass index (BMI), high-density lipoprotein (HDL) cholesterol, total cholesterol, triglycerides, fasting blood glucose, and systolic and diastolic blood pressure levels. We applied generalized estimating equations to account for sibling correlations and adjusted for maternal and participant lifestyle, education, and cardiovascular risk factors. RESULTS: In Third Generation participants (n = 962, mean age = 41 years, 54% were women), 26% of their mothers reported breastfeeding. Compared with non-breastfed individuals, breastfed adult participants had lower multivariable-adjusted BMI (26.1 kg/m2 vs 26.9 kg/m2, P = .04) and higher HDL cholesterol levels (HDL 56.6 mg/dL vs 53.7 mg/dL, P = .01). On additional adjustment for BMI, the association between breastfeeding and HDL cholesterol was attenuated (P = .09). Breastfeeding was not associated with total cholesterol, triglycerides, fasting blood glucose, systolic blood pressure, or diastolic blood pressure. CONCLUSION: Breastfeeding in infancy is inversely associated with adult BMI and positively associated with HDL cholesterol. Associations between breastfeeding and BMI may mediate the association between breastfeeding and HDL cholesterol. PMID- 19559169 TI - Frailty and chronic kidney disease: the Third National Health and Nutrition Evaluation Survey. AB - BACKGROUND: Frailty is common in the elderly and in persons with chronic diseases. Few studies have examined the association of frailty with chronic kidney disease. METHODS: We used data from the Third National Health and Nutrition Examination Survey to estimate the prevalence of frailty among persons with chronic kidney disease. We created a definition of frailty based on established validated criteria, modified to accommodate available data. We used logistic regression to determine whether and to what degree stages of chronic kidney disease were associated with frailty. We also examined factors that might mediate the association between frailty and chronic kidney disease. RESULTS: The overall prevalence of frailty was 2.8%. However, among persons with moderate to severe chronic kidney disease (estimated glomerular filtration rate < 45 mL/min/1.73 m2), 20.9% were frail. The odds of frailty were significantly increased among all stages of chronic kidney disease, even after adjustment for the residual effects of age, sex, race, and prevalent chronic diseases. The odds of frailty associated with chronic kidney disease were only marginally attenuated with additional adjustment for sarcopenia, anemia, acidosis, inflammation, vitamin D deficiency, hypertension, and cardiovascular disease. Frailty and chronic kidney disease were independently associated with mortality. CONCLUSION: Frailty is significantly associated with all stages of chronic kidney disease and particularly with moderate to severe chronic kidney disease. Potential mechanisms underlying the chronic kidney disease and frailty connection remain elusive. PMID- 19559171 TI - Mild hyponatremia carries a poor prognosis in community subjects. AB - OBJECTIVE: Hyponatremia has been shown to predict adverse outcome in congestive heart failure and pneumonia among other common clinical entities, but its significance in the general population is elusive. METHODS: The population-based Copenhagen Holter Study included 671 men and women aged 55 to 75 years with no history of cardiovascular disease, stroke, or cancer. Baseline evaluation included 48-hour ambulatory electrocardiogram monitoring, blood tests, and a questionnaire. Hyponatremia was defined as s-Na < or = 134 mEq/L or s-Na < or = 137 mEq/L according to previously accepted definitions. An adverse outcome was defined as deaths or myocardial infarction. Median follow-up was 6.3 years. RESULTS: Fourteen subjects (2.1%, group A) had s-Na < or = 134 mEq/L, and 62 subjects (9.2%, group B) had s-Na < or = 137 mEq/L. No subject had s-Na < 129 mEq/L. An adverse outcome occurred in 43% of group A, 27% of group B, and 14% of subjects with s-Na >137 mEq/L (controls) (P < .002). Adjusted hazard ratio for adverse outcome was 3.56 (95% confidence interval [CI], 1.53-8.28, P < .005) in group A compared with controls and 2.21 (95% CI, 1.29-3.80, P < .005) in group B after controlling for age, gender, smoking, diabetes, low-density lipoprotein cholesterol, and blood pressure. The hazard ratios were robust for additional adjusting for variables showing univariate association to hyponatremia (ie, beta blocker and diuretic use, heart rate variability, creatinine, C-reactive protein, and NT-pro brain natriuretic peptide). By excluding diuretic users (18% of subjects), the adjusted hazard ratio for adverse outcome was 8.00 (95% CI, 3.04 21.0, P < .0001) in group A and 3.17 (95% CI, 1.76-5.72, P = .0001) in group B compared with controls. CONCLUSION: Hyponatremia is an independent predictor of deaths and myocardial infarction in middle-aged and elderly community subjects. PMID- 19559170 TI - White/black racial differences in risk of end-stage renal disease and death. AB - BACKGROUND: End-stage renal disease disproportionately affects black persons, but it is unknown when in the course of chronic kidney disease racial differences arise. Understanding the natural history of racial differences in kidney disease may help guide efforts to reduce disparities. METHODS: We compared white/black differences in the risk of end-stage renal disease and death by level of estimated glomerular filtration rate (eGFR) at baseline in a national sample of 2,015,891 veterans between 2001 and 2005. RESULTS: Rates of end-stage renal disease among black patients exceeded those among white patients at all levels of baseline eGFR. The adjusted hazard ratios for end-stage renal disease associated with black versus white race for patients with an eGFR > or = 90, 60-89, 45-59, 30-44, 15-29, and <15 mL/min/1.73 m2, respectively, were 2.14 (95% confidence interval [CI], 1.72-2.65), 2.30 (95% CI, 2.02-2.61), 3.08 (95% CI, 2.74-3.46), 2.47 (95% CI, 2.26-2.70), 1.86 (95% CI, 1.75-1.98), and 1.23 (95% CI, 1.12-1.34). We observed a similar pattern for mortality, with equal or higher rates of death among black persons at all levels of eGFR. The highest risk of mortality associated with black race also was observed among those with an eGFR 45-59 mL/min/1.73 m2 (hazard ratio 1.32, 95% CI, 1.27-1.36). CONCLUSION: Racial differences in the risk of end-stage renal disease appear early in the course of kidney disease and are not explained by a survival advantage among blacks. Efforts to identify and slow progression of chronic kidney disease at earlier stages may be needed to reduce racial disparities. PMID- 19559172 TI - Management of small polyps detected by screening CT colonography: patient and physician preferences. AB - BACKGROUND: Management of small polyps found on computed tomography (CT) colonography is controversial and critical to both cancer outcomes and cost. Patient and physician behavior are influenced by personal beliefs and prior experience. Thus, we aimed to understand patient and physician preferences after finding polyps on CT colonography. METHODS: Patients were given a validated handout and survey asking for their preference for evaluation of a "pea-sized" polyp found on CT colonography. By using an Internet survey, physicians were asked how they would manage a 5-mm, 8-mm, or 12-mm polyp, or three 5-mm polyps found by CT colonography in a hypothetical 52-year-old patient of average colorectal cancer risk. Survey reliability was assessed using Cronbach's coefficient alpha. RESULTS: Of the 305 patient respondents, 95% wanted to know if the polyp found on CT colonography was precancerous, 86% stated they would request endoscopic evaluation, and 85% wanted polypectomy. Of the 277 primary care physicians, 71% would refer a 5-mm sigmoid polyp for endoscopy, 86% would refer an 8-mm polyp, 97% would refer a 12-mm polyp, and 91% would refer three 5 mm polyps. Of the 461 gastroenterologists, 83% would refer a 5-mm sigmoid polyp for endoscopy, 96% would refer an 8-mm polyp, 97% would refer a 12-mm polyp, and 93% would refer three 5-mm polyps. Overall, 75% of physicians indicated the fear of missing a precancerous lesion would prompt referral for colonoscopy. CONCLUSION: Both patients and physicians overwhelmingly preferred to follow up small polyps identified by CT colonography with endoscopy, suggesting that population-based CT colonography screening programs in which polyps are not removed might require significant patient and physician education before implementation. PMID- 19559174 TI - The modern teaching physician--responsibilities and challenges: an APDIM white paper. PMID- 19559173 TI - Herpes simplex type-2 meningitis: presentation and lack of standardized therapy. AB - BACKGROUND: Herpes simplex type-2 (HSV-2) causes both primary and recurrent lymphocytic meningitis, but optimal patient management is not well defined. METHODS: In this retrospective observational study, we reviewed the medical records of patients with HSV-2-positive cerebrospinal fluid samples in our laboratory between January 2001 and January 2005. RESULTS: During the study period, 23 patients, aged 16 to 83 years, had HSV-2 detected in spinal fluid. Nineteen (83%) had meningitis and 4 (17%) had evidence of meningoencephalitis. Seventy-four percent were female. Two (8.7%) had a history of prior genital herpes, and one (4.3%) had genital lesions noted at the time of presentation. Genital examinations were performed at presentation in only 3 patients. Seven (30.4%) patients reported previous episodes of meningitis. Two celibate women developed HSV-2 meningitis or meningoencephalitis following lumbar steroid injection for spinal stenosis. One woman developed HSV-2 meningoencephalitis 3 days postpartum following cesarean section. Antiviral treatment for uncomplicated HSV-2 meningitis varied from none (4 patients) to 14-21 days of intravenous (IV) acyclovir therapy (4 patients). The 11 remaining patients with meningitis received 1-7 days of IV therapy, followed by 7-21 days of oral antiviral therapy. Three of 4 patients with meningoencephalitis received 21 days of IV acyclovir, and one received 3 days IV acyclovir followed by 14 days of oral therapy. CONCLUSIONS: HSV-2 meningitis presents most often without a history of genital herpes, recurrent meningitis, or genital symptoms. Current management practices are highly variable and may lead to unnecessary hospitalization and prolonged intravenous therapy. PMID- 19559175 TI - Reconstruction of two independent neo-atria after resection of recurrent leiomyosarcoma. AB - We describe a case of a patient with recurrent leiomyosarcoma involving both atria after a previous right pneumonectomy. The patient was treated with wide resection of the mass and separate reconstruction of the cardiac cavities with prosthetic material. PMID- 19559176 TI - Aortic valve replacement in a patient with Osler-Rendu-Weber disease. AB - Osler-Rendu-Weber (hereditary hemorrhagic telangiectasia) disease is an uncommon disease characterized by the presence of abnormal telangiectasias and arteriovenous malformations that cause recurrent episodes of bleeding. We present a patient with Osler-Rendu-Weber disease, with a history of multiple major bleeding events and severe aortic valve stenosis, who underwent aortic valve replacement. Unexpectedly, the postoperative course was uneventful, and there was no untoward bleeding in the early or in the late postoperative follow-up. PMID- 19559177 TI - Ossifying thymoma clinically presenting with peripheral T-cell lymphocytosis. AB - We believe there has been only one ossifying thymoma reported in the English literature. We herein reported another such case with additional peculiar presentation of peripheral T-cell lymphocytosis. A 62-year-old woman was incidentally found to have an anterior mediastinal tumor during a medical check up, which was surgically resected 42 months later and histopathologically confirmed to be a type B1 thymoma with stromal ossification. Fifty months after tumor removal, this patient remains alive and well without relapsed disease. PMID- 19559178 TI - Aberrant right subclavian artery aneurysm in coexistence with a common carotid trunk. PMID- 19559179 TI - Ethical obligation of surgeons to noncompliant patients: can a surgeon refuse to operate on an intravenous drug-abusing patient with recurrent aortic valve prosthesis infection? PMID- 19559180 TI - Cerebrospinal fluid drainage during thoracic aortic repair: safety and current management. AB - BACKGROUND: The benefit of cerebrospinal fluid (CSF) drainage during thoracic aortic repair has been established. Few studies, however, report management and safety of CSF drainage. METHODS: Between September 1992 and August 2007, 1,353 repairs of the thoracic aorta were performed, with 82% using CSF drainage. The CSF drainage was not used in cases of rupture, acute trauma, infection, or prior paraplegia. Thirty-one percent (76 of 246) of patients without CSF drainage were repaired prior to standardized use. All drains were inserted by cardiovascular anesthesia staff. Repairs were performed using distal aortic perfusion with heparinization. Early management involved free drainage to maintain CSF pressure less than 10 mm Hg, but was later modified to limit CSF drainage unless neurologic deficit occurred. RESULTS: Cerebrospinal fluid drainage was technically achieved in 99.8% (1,105 of 1,107) of cases. The CSF catheter-related complications occurred in 1.5% (17 of 1,107) of patients. No spinal hematomas were observed. The CSF leaks with spinal headache, CSF leak without spinal headache, spinal headache, intracranial hemorrhage, catheter fracture, and meningitis occurred in 6 (0.54%), 1 (0.1%), 2 (0.2%), 5 (0.45%), 1 (0.1%), and 2 (0.2%) cases, respectively. Mortality from subdural hematoma was 40% (2 of 5), and from meningitis was 50% (1 of 2). Spinal headaches resolved with conservative management. All CSF leaks resolved, but 71% (5/7) required blood patches. Since implementation of a limited CSF drainage protocol, no subdural hematomas have been observed. CONCLUSIONS: Cerebrospinal fluid drainage for thoracic aortic repairs can be performed safely with excellent technical success. Perioperative management of CSF drains requires diligent monitoring and judicious drainage. Standardizing CSF management may be beneficial. PMID- 19559182 TI - Invited commentary. PMID- 19559181 TI - Arch aneurysm repair with long elephant trunk: a 10-year experience in 111 patients. AB - BACKGROUND: We studied the long-term outcome of arch aneurysm repair with a long elephant trunk (LET) anastomosed at the base of brachiocephalic artery. METHODS: Between 1998 and 2008, 111 patients underwent arch aneurysm repair with LET. A 4 branched graft was sutured to the sinotubular junction, the distal ascending aorta transected, and a LET inserted into the aortic arch while selective cerebral perfusion was maintained. The graft distal end was anastomosed to the LET, incorporating the distal ascending aorta, and arch vessels were anastomosed to graft branches. RESULTS: Concomitantly, 33 patients (30%) underwent other cardiac procedures, including 11 aortic root replacements. Two patients died (1.8%) within 30 days and 7 died (6.3%) after 30 days. Perioperative morbidity included 2 (1.8%) with stroke, 3 (2.7%) with paraplegia, and 1 (0.9%) with paraparesis. Postoperative computed tomography scans revealed complete aneurysmal thrombosis around the LET in 88 patients (79%), who were monitored without a second-stage procedure. Among 23 patients with incomplete thrombosis, 19 underwent a second-stage procedure to complete distal fixation of the LET. Overall survival was 88%, 83%, and 75%, at 1, 3, and 5 years after aneurysm repair with the LET. No aneurysm rupture or reexpansion occurred in patients with complete thrombosis. Four patients with incomplete thrombosis died of rupture before the second-stage procedure. CONCLUSIONS: Our results demonstrated safety and good durability of the LET technique and suggest that this technique is a simple and safe procedure that is applicable to a variety of arch aneurysms. PMID- 19559183 TI - Long-term patency of 1108 radial arterial-coronary angiograms over 10 years. AB - BACKGROUND: To avoid late vein graft atheroma and failure, we have used arterial grafts extensively in coronary operations. The radial artery (RA) is the conduit of second choice. This study determined the long-term patency of the RA as a coronary graft. METHODS: Two independent observers evaluated 1108 consecutive postoperative RA conduit angiograms performed between January 1997 and June 2007 for cardiac symptoms. Mean time to postoperative angiography was 48.3 months (range, 1 to 132 months). An RA graft was considered failed (nonpatent) if there was stenosis exceeding 60%, string sign, or occlusion. Patency was determined over time, by coronary territory grafted and by the degree of native coronary artery stenosis (NCAS). RESULTS: At a mean of 48.3 months, 982 of the 1108 RA grafts (89%) were patent. RA patencies for the left anterior descending were 96% (24 of 25), diagonal/intermediate, 90% (121 of 135); circumflex marginal, 89% (499 of 561); right coronary, 83% (38 of 46); posterior descending, 89% (253 of 286); and left ventricular branch/posterolateral, 86% (47 of 55). Patency was 87.5% (56 of 64) for NCAS of less than 60% compared with 89% (926 of 1044; p = 0.89) for NCAS exceeding 60%. Of 318 RAs in place more than 5 years, 294 (92.5%) were patent, and for 107 RAs in place for more than 7 years, 99 were patent (92.5%). Patency was consistent through each year of the decade. Mechanisms of failure did not involve development of atherosclerosis. Patent RA grafts were smooth, with no angiographic evidence of atheroma. CONCLUSIONS: Late patencies of RA grafts are excellent and justify continuing use of the RA in coronary operations. PMID- 19559184 TI - Sequential radial artery grafts for multivessel coronary artery bypass graft surgery: 10-year survival and angiography results. AB - BACKGROUND: Increasing the number of arterial grafts for coronary artery bypass grafting (CABG) has been linked to improved late survival. Currently, it is not known if these long-term benefits are also true when sequential radial artery (RA) grafts are the primary means to maximizing arterial revascularization. METHODS: We compared late survival of 532 consecutive patients receiving sequential RA grafts (sequential RA group: 438 men; 462 with three-vessel disease) with that of a 4,131 contemporaneous internal thoracic artery (ITA) with saphenous vein (SV) multivessel CABG cohort (conventional group). Graft failure rates were determined from symptom-driven repeat angiography films in 122 sequential RA patients performed 2 to 4,317 days after surgery. Median survival sequential RA follow-up was 5.3 years (range, 0.5 to 12.3). RESULTS: The sequential RA patients received a total of 1,181 RA grafts (538 sequential [30 triple] and 75 single) along with 636 SV and 533 ITA. Overall RA graft failure (80 of 272; 29%) was intermediate to that for ITA (7 of 121; 5.8%; p < 0.001) and vein (54 of 133, 41.6%; p = 0.032) grafts. Sequential versus nonsequential RA failure did not differ (77 of 252 [31%] versus 3 of 20 [15%]; p = 0.202), while failure of the proximal (36 of 123; 29%) and distal (40 of 129; 31%) components of sequential RA grafts were essentially identical. A total of 69 deaths (6 operative; 1.1%) have occurred in the sequential RA cohort. Unadjusted 10-year sequential RA cohort survival was 76.2% overall, and 79.0% for the 454 primary isolated CABG subgroup. The risk-adjusted 10-year survival using a logit propensity score was substantially better for the sequential RA cohort versus the conventional CABG cohort (risk ratio [95% confidence interval] 0.61 [0.44 to 0.85]; p = 0.003). CONCLUSIONS: Sequential RA grafting is a safe method for maximizing arterial revascularization and is associated with excellent 10-year survival that seems to be superior to conventional or ITA/SV CABG results. Also, the similar proximal and distal sequential RA patency mitigates concerns of a clinically significant effect of increased vasoreactivity of distal segments of RA conduits. PMID- 19559185 TI - Invited commentary. PMID- 19559186 TI - Additive costs of postoperative complications for isolated coronary artery bypass grafting patients in Virginia. AB - BACKGROUND: Complications after open-heart surgery result in an increased length of stay and greater financial burdens for all. The purpose of this study was to measure the additive costs of postoperative complications for selected subgroups of patients after coronary artery bypass grafts in the Commonwealth of Virginia. METHODS: A multiyear statewide data repository with clinical and billing data was used to measure outcomes for the period 2004 to 2007. The Society of Thoracic Surgeons records matched with Universal Billing (UB-04) charge data for all payers were used to estimate the additive costs of cardiac surgical outcomes using cost-to-charge ratios. Additive cost was defined as the difference between the baseline cost of an average case with no complications and one with a postoperative morbidity or mortality. Multivariate analysis was used to account for important covariates and apportion incremental costs. RESULTS: The baseline cost of isolated coronary artery bypass grafting (CABG) cases with no complications during the study period was $26,056. Isolated atrial fibrillation was the most frequently cited complication and had the lowest additive cost ($2,574). Additive costs for isolated CABG patients were greatest for those cases involving prolonged ventilation ($40,704), renal failure ($49,128), mediastinitis ($62,773), and operative mortality ($49,242). CONCLUSIONS: Additive costs can serve as an indicator for pursuing quality improvement initiatives. Our results suggest additive costs vary according to type of postoperative complication and comorbidities. Regional collaborations of multidisciplinary groups in cardiac surgery are an effective means to implement quality guidelines and drive down additive costs. PMID- 19559188 TI - Invited commentary. PMID- 19559187 TI - In-hospital outcomes of off-pump multivessel total arterial and conventional coronary artery bypass grafting: single surgeon, single center experience. AB - BACKGROUND: Despite increasing recognition that off-pump coronary artery bypass surgery and total arterial revascularization individually are associated with improved outcomes, concerns persist regarding the safety of combining these two techniques. We compared in-hospital outcomes for off-pump multivessel total arterial and conventional coronary artery bypass grafting. METHODS: From September 1998 to September 2008, 580 consecutive patients receiving off-pump multivessel arterial grafts only were compared with a control group of patients (n = 806) undergoing off-pump coronary artery bypass grafting with internal thoracic artery and saphenous veins operated on by the same surgeon. Two different statistical approaches were used to compare groups in this retrospective analysis. First, propensity score analysis was used to match patients from each group. Second, a multivariate analysis was performed looking at a combined patient outcome of death, intraaortic balloon counterpulsation utilization, myocardial infarction, stroke, prolonged ventilation, and reoperation for any cause on all patients in both groups. RESULTS: After matching by propensity score, the major clinical outcomes in total arterial (n = 346) and control (n = 346) groups were found to be similar. The in-hospital mortality in the total arterial group was 1.2% as compared with 2.0% in matched patients (p = 0.8). However, patients in the total arterial group were found to have a significantly increased incidence of reexploration for bleeding (p < 0.0001) and blood product usage (p < 0.0001). There was a higher incidence of combined morbidity outcome (18.8% versus 12.1%; p = 0.001) for the control group compared with the total arterial group. Multivariate analysis failed to show that total arterial grafting was an independent predictor of the combined morbidity outcome. CONCLUSIONS: Off-pump multivessel total arterial grafting can be performed safely with superior in-hospital outcomes compared with off-pump conventional coronary artery bypass grafting. PMID- 19559189 TI - Remodeling of reconstructed left anterior descending coronary arteries with internal thoracic artery grafts. AB - BACKGROUND: The internal thoracic artery (ITA) remodels its diameter in response to flow requirements. The objective of this study was to elucidate the remodeling capacity of the reconstructed coronary artery using the ITA. METHODS: We evaluated coronary angiograms in 63 patients who had left anterior descending artery (LAD) segmental reconstruction with or without endarterectomy after off pump coronary artery bypass graft surgery. The diameters of the ITA and reconstructed coronary artery were measured early and at 1 year after surgery. RESULTS: The mean diameter of the reconstructed LAD was significantly larger than that of the ITA, but significantly decreased 1 year after surgery (2.69 +/- 0.53 mm versus 1.87 +/- 0.39 mm; p > 0.0001). The proximal ratio, the ratio of the ITA to proximal reconstructed coronary artery, and the distal ratio, the ratio of the distal LAD to distal reconstructed coronary artery, increased to a value of almost 1.0 (0.77 +/- 0.11 versus 1.05 +/- 0.18, p < 0.0001, and 0.77 +/- 0.14 versus 0.92 +/- 0.12, p < 0.0001, respectively). Based on the mean diameter of the reconstructed coronary artery, there were no relationships between the use of endarterectomy and the degree of native coronary stenosis. The proximal ratio in the group with severe stenosis was significantly greater than that in the group with mild stenosis (1.08 +/- 0.18 versus 0.95 +/- 0.16; p = 0.036), although the distal ratio was not different between the two groups. CONCLUSIONS: Vascular remodeling of the coronary artery reconstructed with the ITA is observed within 1 year after surgery. PMID- 19559190 TI - Invited commentary. PMID- 19559191 TI - Clopidogrel and aspirin versus clopidogrel alone on graft patency after coronary artery bypass grafting. AB - BACKGROUND: Clopidogrel and aspirin are the most popular antiplatelet agents for anticoagulation management after coronary artery bypass grafting (CABG) in clinical practice, but there is neither a standard antiplatelet therapy for patients undergoing CABG, nor an exact conclusion about its effects on graft patency until now. METHODS: One-hundred and ninety-seven selected patients undergoing CABG were assigned to two groups according to antiplatelet drug: the clopidogrel group of 102 patients who received clopidogrel (75 mg) daily; and the combination group of 95 patients who received clopidogrel (75 mg) plus aspirin (100 mg) daily. Multislice computed tomography angiography was performed to evaluate graft patency at 1 month and 12 months after CABG. RESULTS: There were no significant differences between the two groups in preoperational data. At 1 month and 12 months after CABG graft patency rates of clopidogrel group were, respectively, 99.0% and 96.9% for the left internal mammary artery (LIMA) and 98.1% and 93.5% for the saphenous vein grafts; those of the combination group were, respectively, 98.9% and 97.8% for LIMA, and 98.2% and 96.3% for saphenous vein grafts. There were no significant differences in graft patency between the two groups (p > 0.05). CONCLUSIONS: Either clopidogrel plus aspirin or clopidogrel alone maintain high graft patency in the early postoperative phase after CABG. The observed trend toward higher patency rates in patients treated with clopidogrel plus aspirin compared to those in the clopidogrel group did not reach statistical significance. PMID- 19559192 TI - Invited commentary. PMID- 19559193 TI - Survival of cardiorespiratory arrest after coronary artery bypass grafting or aortic valve surgery. AB - BACKGROUND: Study objectives were to (1) report the clinical profile of and outcome for patients who experience a cardiorespiratory arrest after coronary artery bypass grafting or aortic valve replacement, and (2) identify factors associated with improved probability of survival. METHODS: We identified 108 consecutive patients who had cardiorespiratory arrest after coronary artery bypass grafting or aortic valve replacement between April 1999 and June 2008. We studied the characteristics of arrests and survivors, and performed a multivariate logistic analysis to determine features associated with survival to hospital discharge. RESULTS: Cardiac arrest (n = 86) was more common than respiratory arrest (n = 13; unknown cause, n = 9). Cardiorespiratory arrest occurred with decreasing frequency from the day of surgery. Ventricular fibrillation or tachycardia was the dominant mechanism of cardiac arrest (70% versus 17% for asystole versus 13% for pulseless electrical activity), and the principal causes were postoperative myocardial infarction (n = 46; 53%) and tamponade or bleeding (n = 21; 24%). Resternotomy was performed in 45 patients (52%), cardiopulmonary bypass reinstituted in 14 (16%), and additional grafts constructed in 5 (6%). The causes of respiratory arrest were mainly pulmonary (n = 8) and neurologic (n = 5). Survival to hospital discharge was better for respiratory arrest (69%) than for cardiac arrest (50%). Older age, ejection fraction less than 0.30, and postoperative myocardial infarction decreased the probability of survival. CONCLUSIONS: Ventricular fibrillation or tachycardia was the most common mechanism, and myocardial infarction, the predominant precipitating cause of cardiac arrest after coronary artery bypass grafting or aortic valve replacement. Despite aggressive resuscitation, outcome is poor. Young patients with good left ventricular function had a better probability of survival if they did not suffer a postoperative myocardial infarction. PMID- 19559194 TI - Invited commentary. PMID- 19559196 TI - Invited commentary. PMID- 19559195 TI - The July effect: impact of the beginning of the academic cycle on cardiac surgical outcomes in a cohort of 70,616 patients. AB - BACKGROUND: Because surgical residents' level of experience may be at its nadir early in the academic year, academic seasonality-or the "July effect"-could affect cardiac surgical outcomes. METHODS: Prospectively collected data from the Department of Veterans Affairs Continuous Improvement in Cardiac Surgery Program were used to identify 70,616 consecutive cardiac surgical procedures performed between October 1997 and October 2007. Morbidity and mortality rates were compared between early (July 1 to August 31, n = 11,975) and late (September 1 to June 30, n = 58,641) periods in the academic year. A prediction model was constructed by using stepwise logistic regression modeling. RESULTS: The two patient groups had similar demographic and risk variables. Isolated coronary artery bypass grafting accounted for 76.7% of early-period procedures and 75.8% of later-period procedures (p = 0.03). Morbidity rates did not differ significantly between the early (14.0%) and later periods (14.2%; odds ratio [OR], 1.01; 95% confidence interval [CI], 0.96 to 1.07; p = 0.67) and operative mortality was similar, 3.7% vs 3.9% (OR, 0.99; 95% CI, 0.89 to 1.11; p = 0.90). The early portion of the year was associated with longer cardiac ischemia times (84 +/- 40 vs 83 +/- 42 minutes), cardiopulmonary bypass times (126 +/-52 vs 124 +/-56 minutes), and total surgical times (295 +/- 90 vs 288 +/- 90 minutes; p < 0.05 for all). CONCLUSIONS: The early part of the academic year was associated with slightly longer operative times; however, risk-adjusted outcomes were similar in both periods. This finding should lessen concerns about the quality of cardiac surgical care at the beginning of the academic year. PMID- 19559197 TI - Is prosthetic anuloplasty necessary for durable mitral valve repair? AB - BACKGROUND: Because emerging percutaneous mitral valve repair may address only leaflets and not the anulus, we compared durability of mitral valve repair with and without prosthetic anuloplasty. METHODS: From 1985 to 2007, 3,057 patients underwent primary isolated posterior leaflet repair for degenerative mitral disease either with prosthetic anuloplasty (n = 2,754, 90%) or without (n = 303, 9.9%: no anuloplasty, 68; suture anuloplasty, 7; pericardial anuloplasty, 228). Most of the latter operations occurred in the early 1990s. Differences in patient characteristics were addressed by propensity-score adjustment and matching (214 pairs). In all, 3,870 echocardiograms for 1,236 patients were available for assessing mitral regurgitation after prosthetic anuloplasty and 257 in 99 patients without one. Mean follow-up for mitral valve reoperation was 4.2 +/- 4.1 years, with 13,003 patient-years of data available for analysis. RESULTS: Early, and to a lesser degree late, postoperative mitral regurgitation was less after prosthetic anuloplasty than repair without one, and this difference persisted after risk adjustment and in propensity-matched patients (p = 0.0002). Freedom from mitral valve reoperation was 96% and 94% at 10 years after repair with versus without prosthetic anuloplasty in unmatched groups, and 97% and 96% in matched groups (p = 0.3), respectively. Unadjusted survival was greater with than without prosthetic anuloplasty (84% versus 81% at 10 years, p = 0.009), but similar after propensity adjustment and in matched pairs. CONCLUSIONS: Mitral valve repair without a prosthetic anuloplasty was associated with accelerated return of mitral regurgitation, although risk-adjusted survival was similar. This finding has important implications for durability of percutaneous mitral repair techniques that do not address both leaflets and anulus. PMID- 19559198 TI - Long-term results of the leaflet extension technique in aortic regurgitation: thirteen years of experience in a single center. AB - BACKGROUND: We evaluated the effectiveness and durability of the leaflet extension technique for correction of aortic regurgitation (AR) and the long-term clinical results. METHODS: Between March 1995 and August 2004, 41 consecutive patients were included. The mean age was 32.2 +/- 13.9 years. The causes of AR were rheumatic in 31 patients (75.5%), degenerative in 2 patients (4.9%), bicuspid aortic valve in 4 patients (9.8%), infective endocarditis in 1 patient (2.4%), and congenital in 3 patients (7.3%). Leaflet extensions were performed in three leaflets for 32 patients, two leaflets for 3 patients, and only one leaflet for 6 patients. The mean follow-up duration was 92.9 +/- 48.4 months. RESULTS: There were no early deaths and 2 late deaths. One patient died of cancer and the other patient died of infective endocarditis. The cardiac-related mortality was 2.4% (1 of 41 patients). During a mean follow-up of 7 years, severe AR was detected in 1 patient and moderate AR in 6 patients (17.0%; 7 of 41 patients). The causes of recurrent AR were infective endocarditis in 3 patients, disease progression in 3 patients, and Behcet's diseases in 1 patient. We performed 6 reoperations (14.6%), 3 in patients owing to infective endocarditis, 2 in patients owing to disease progression, and 1 in a patient owing to the suture dehiscence associated with Behcet's disease. The cumulative survival was 92.6% at 13 years. Freedom from recurrent AR was 97.5% at 5 years, 81.7% at 10 years, and 68.1% at 13 years. CONCLUSIONS: The long-term durability of the leaflet extension technique was acceptable. The reoperations increased with time, but pericardial leaflet dysfunction was not the cause. PMID- 19559199 TI - Invited commentary. PMID- 19559200 TI - Unexpected complications of transapical aortic valve implantation. AB - BACKGROUND: Recent series have reviewed the results of transapical aortic valve implantation (TAVI). However, specific problems of this new procedure are not well-described. Unexpected complications due to the procedure and their management are reported. METHODS: Eighteen patients underwent TAVI using the Edwards Sapien bioprosthesis (Edwards Lifesciences Inc, CA) between September 2007 and June 2008 due to contraindications of conventional surgery (n = 5) or high operative risk (n = 13). The system was introduced through 2 purse string sutures in the apex under echocardiographic and fluoroscopic control. RESULTS: The implantation success rate and initial procedural success were 100%. There was no intraoperative death and no stroke. During the procedure, two cases of ventricular fibrillation consequent to rapid pacing were treated by cardioversion. Acute mitral regurgitation due to traction of the subvalvular apparatus by the guidewire and acute aortic regurgitation from pressure on a bioprosthesis cusp by the guidewire were diagnosed by transesophageal echocardiography and reversed by the removal of the guidewire. Another case of aortic regurgitation was due to incomplete deployment of the bioprosthesis and was managed by a "valve after valve" procedure. Two patients died on postoperative day 2 from left ventricular failure. In one patient the postmortem study showed, despite correct implantation of the bioprosthesis, a hematoma of the septum with a small ventricular septal defect. The total in-hospital death was 27.7% (5 patients). There was no periprocedural bleeding but in one patient delayed rupture of the apex (36 hours after the procedure) necessitated emergency surgery. A false aneurysm of the apex appeared 3 months after surgery in another patient. Closure of the apex was performed through sternotomy and cardiopulmonary bypass with an uneventful follow-up. CONCLUSIONS: The TAVI is associated with incidents and complications different to those encountered in conventional aortic valve surgery. Recognizing their existence contributes to elucidating their mechanisms and to propose solutions to avoid or treat them. PMID- 19559201 TI - Invited commentary. PMID- 19559203 TI - The Cox-Maze III procedure success rate: comparison by electrocardiogram, 24-hour holter monitoring and long-term monitoring. AB - BACKGROUND: The detection of atrial arrhythmia recurrence is more accurate when using long-term (5 days to 3 weeks) continuous monitoring devices. In this study, we focus on the comparison of the recurrence of atrial arrhythmias in patients after the Cox-Maze III procedure obtained by three modalities: electrocardiography (ECG), 24-hour Holter monitoring, and long-term monitoring (LTM). METHODS: Patients with follow-up longer than 6 months who reported sinus rhythm while not taking antiarrhythmic drugs were eligible. Atrial arrhythmias longer than 30 s were considered a recurrence. The ECG, 24-hour Holter monitoring, and LTM (5 days) reports were ascertained and compared at the same time. RESULTS: Patients (n = 291) underwent the full Cox-Maze III procedure, with 194 eligible for the study and 76 agreeing to participate. The average time to monitoring after surgery was 9.8 (+/- 7.7) months. The ECGs determined 96% of patients in sinus rhythm, Holter monitoring determined 91% in sinus rhythm, and LTM indicated 84% in sinus rhythm. Comparing ECG results and LTM results revealed that 9 patients (12%) had a significant rhythm change. Holter monitoring did not capture all the patients having events lasting longer than 1 hour. No additional information was captured by the use of LTM in patients with paroxysmal atrial fibrillation. CONCLUSIONS: This study reconfirmed that ECG overestimated the success rate after the Cox-Maze III operation by 12% compared with LTM. These changes may carry clinical significance when determining the success of the Cox Maze III procedure and determining the medical management, including antiarrhythmic and anticoagulation therapy, of the patients who were found to have significant events. PMID- 19559202 TI - The effect on long-term survival of erythrocyte transfusion given for cardiac valve operations. AB - BACKGROUND: Studies in patients undergoing coronary artery bypass grafting (CABG) have shown an increased long-term mortality rates associated with perioperative blood transfusions. However, some studies in other patient populations have shown no effect on death or even a lowered mortality rate in patients receiving blood transfusions, which suggests that the effects of blood transfusion may be disease dependent. METHODS: Data of all patients who underwent valve operations with or without associated CABG between October 2, 1991, and November 14, 2007, were obtained from the department's database and analyzed using logistic regression for 30-day and Cox models for long-term mortality to determine the effects of transfusion on death. To control for the potential interaction between transfusion and complications and sicker patients being more likely to receive blood, we separately analyzed the data for the different valve populations and used propensity analysis to control for sicker patients being more likely to receive blood. RESULTS: Of 1823 patients who underwent valve operations, the operation was isolated in 993 and combined with CABG in 830. By 30 days, 125 patients (6.9%) had died, and 717 (39%) were dead at follow-up. After controlling for type of operation and factors that influenced the transfusion decision, transfusion was associated with increased death only in patients who had combined valve and CABG, and not in isolated valve operations. CONCLUSIONS: Transfusion had no effect on the mortality rate after isolated valve operations but was associated with increased mortality when valve operations were combined with CABG. PMID- 19559205 TI - Invited commentary. PMID- 19559204 TI - Current use of prophylactic strategies for postoperative atrial fibrillation: a survey of Canadian cardiac surgeons. AB - BACKGROUND: Evidence from multiple trials demonstrates the efficacy of prophylactic beta-blocker, amiodarone, and corticosteroid administration in reducing the incidence of postoperative atrial fibrillation. Despite this information, these interventions remain infrequently or inappropriately utilized. This study was designed to assess the frequency with which these prophylactic strategies are currently being used and to identify concerns and barriers to more widespread application. METHODS: A link to an online survey was e-mailed to all practicing cardiac surgeons in Canada. Each surgeon was given a unique log-in identification number to complete the survey online through a secure web page. RESULTS: Surveys were sent to 166 surgeons; 119 completed surveys (72%) were returned. Only 58% of respondents routinely use beta-blockade for prophylaxis. For nonusers, 44% are unconvinced of the evidence for this practice. The routine use of amiodarone among surgeons was 19%. Of the remainder, 43% cited a perceived increased risk of complications as the reason for not using this therapy. An additional 29% considered the therapy was excessively complicated or time consuming. Corticosteroids were routinely used by only one surgeon. Major barriers to use of steroids were unconvincing evidence (76%), a perceived increased risk of wound infection (38%), and hyperglycemia (30%). CONCLUSIONS: Despite level 1 evidence, the use of beta-blockers, amiodarone, and corticosteroids for prophylaxis of atrial fibrillation among Canadian surgeons remains less than expected. The results of this survey support the need for further clinical trials with robust and clinically relevant outcomes that may further influence surgeons to adopt this practice. PMID- 19559206 TI - Nontraditional surgical approaches for implantation of pacemaker and cardioverter defibrillator systems in patients with limited venous access. AB - BACKGROUND: Indications for placement of implantable cardioverter-defibrillators (ICD) and pacemakers have expanded, and traditional transvenous implantation may not be feasible in patients with aberrant anatomy or venous obstruction. In these settings, successful lead placement has required innovative surgical approaches. A case series of successful placement of these systems in challenging patients is presented. METHODS: A 2-year retrospective study of patients undergoing placement of minimally invasive epicardial pacing leads or ICD coils was performed. RESULTS: Eleven patients underwent minimally invasive surgical placement of leads or coils. None were converted to open sternotomy. One required extension to minianterior thoracotomy. Causes of intravenous placement failure included aberrant anatomy with failure to access coronary sinus in 9 and venous occlusion in 2. Four patients had previous operations through a median sternotomy. Procedures included left video-assisted thoracoscopic (VATS) placement of a left ventricular epicardial lead in 8, left VATS conversion to minianterior thoracotomy left ventricular epicardial lead placement in 1, left VATS placement of ICD coil in 1, subxiphoid placement of a right ventricular epicardial lead in 1, subxiphoid ICD coil in 2, and subcutaneous ICD coil placement in 3. Mean hospitalization was 4.6 days. Postoperative hypotension and pulmonary edema occurred in 27% of patients. No patients died. CONCLUSIONS: Conventional transvenous lead implantation may be difficult or impossible in some patients with aberrant or occluded venous access. Novel surgical approaches with the use of minimally invasive procedures can establish optimally functional pacing and ICD systems without sternotomy and low associated morbidity. PMID- 19559207 TI - Cardiac vagal stimulation eliminates detrimental tachycardia effects of dobutamine used for inotropic support. AB - BACKGROUND: Many patients require temporary inotropic support after cardiac surgery, and dobutamine is one of the commonly used drugs for this purpose. However, dobutamine infusion is frequently associated with unwanted sinus tachycardia. Selective sinus node electrical vagal stimulation through a discrete epicardial ganglionic plexus (fat pad) approach can achieve sinus rate slowing. Because sinus node fat pad vagal stimulation (SNFP-VS) can easily be applied during or after cardiac surgery, we hypothesized that combining selective SNFP-VS with dobutamine could produce desired hemodynamic improvement while avoiding sinus tachycardia in patients when inotropic drug support is needed. METHODS: This exploratory experimental study was performed in 7 open-chest dogs. Dobutamine (2.5 to 10 microg x kg(-1) x min(-1)) was infused at a rate producing at least 30% increase in sinus rate and cardiac output. Then electrical SNFP-VS was applied in the epicardial ganglionic plexus located at the right pulmonary vein-atrial junction, to slow the sinus rate back to control level. Hemodynamic data during control, with steady-state dobutamine infusion, and with dobutamine plus SNFP-VS were collected and compared. RESULTS: Dobutamine significantly increased heart rate, systolic and diastolic blood pressures, peak left ventricular systolic pressure, positive and negative maximal derivatives of left ventricular pressure, and cardiac output. Combining SNFP-VS with dobutamine eliminated sinus rate increase while preserving all major hemodynamic benefits. Selective SNFP-VS itself had no direct effect on cardiac contractility during atrial pacing. CONCLUSIONS: Combining SNFP-VS with dobutamine could achieve hemodynamic improvement while avoiding sinus tachycardia in this dog model, suggesting that similar strategy may also be applied in patients. PMID- 19559208 TI - Invited commentary. PMID- 19559209 TI - Novel biomarkers early predict the severity of acute kidney injury after cardiac surgery in adults. AB - BACKGROUND: The purpose of this study was to investigate the ability of neutrophil gelatinase-associated lipocalin (NGAL), cystatin C, and their combination in predicting the duration and severity of acute kidney injury (AKI) after cardiac surgery in adults. METHODS: Using data from a prospective observational study of 100 adult cardiac surgical patients, we correlated early postoperative concentrations of plasma NGAL and serum cystatin C with the duration (time during which AKI persisted according to the Acute Kidney Injury Network criteria) and severity of AKI (change in serum creatinine) and with length of stay in intensive care. RESULTS: We found a mean AKI duration of 67.2 +/- 41.0 hours which was associated with prolonged hospitalization (p < 0.001). NGAL, cystatin C, and their combination on arrival in intensive care correlated with subsequent AKI duration (all p < 0.01) and severity (all p < 0.001). The area under the receiver operating characteristic curve for AKI prediction was 0.77 (95% confidence interval: 0.63 to 0.91) for NGAL and 0.76 (95% confidence interval: 0.61 to 0.91) for cystatin C on arrival in intensive care. Both markers also correlated with length of stay in intensive care (p = 0.037; p = 0.001). Neutrophil gelatinase-associated lipocalin and cystatin C were independent predictors of AKI duration and severity and of length of stay in intensive care (all p < 0.05). The value of cystatin C on arrival in intensive care appeared to be due to a carry-over effect from preoperative values. CONCLUSIONS: Immediately postoperatively, NGAL and cystatin C correlated with and were independent predictors of duration and severity of AKI and duration of intensive care stay after adult cardiac surgery. The combination of both renal biomarkers did not add predictive value. PMID- 19559210 TI - Invited commentary. PMID- 19559211 TI - Mid-term outcomes in adults with ebstein anomaly and cavopulmonary shunts. AB - BACKGROUND: In patients with Ebstein anomaly and poorly functioning right ventricles, a cavopulmonary shunt (CPS) can be created to reduce the preload on the right ventricle. The purpose of this study was to examine the early and mid term outcomes in adults with Ebstein anomaly who have undergone tricuspid valve repair or replacement with or without a concomitant CPS. METHODS: We examined the outcomes of 40 consecutive patients seen at our center with Ebstein anomaly who had undergone tricuspid valve repair or replacement with (n = 23) or without (n = 17) concomitant CPS. Follow-up data were obtained by either chart review or contacting the referring cardiologist. Mid-term survival was examined using Kaplan-Meier curves. RESULTS: The mean age at surgery was similar in patients with and without CPS (42 +/- 12 versus 39 +/- 19 years; p = 0.63). There were 2 early postoperative deaths owing to refractory right-sided heart failure. Mid term follow-up data were available in 95% of patients. The mean follow-up time was 6.7 +/- 4.8 years. Patients who received a CPS more commonly had preoperative heart failure or cyanosis (p = 0.04) and had worse preoperative functional status (p = 0.09). In both groups, arrhythmias were the most common late complication. There were 5 late deaths, 3 of which occurred in patients with CPS. Five-year survival with or without CPS was comparable (83% +/- 9% versus 86% +/- 10%; p = 0.85). CONCLUSIONS: Adolescent and adult patients with Ebstein anomaly undergoing tricuspid valve replacement or repair and concomitant CPS are at risk for early and mid-term complications. However, Ebstein surgery along with CPS appears to be a reasonable surgical strategy in patients not thought to be suitable for tricuspid valve surgery alone. PMID- 19559212 TI - The rastelli procedure for transposition of the great arteries: resection of the infundibular septum diminishes recurrent left ventricular outflow tract obstruction risk. AB - BACKGROUND: The Rastelli procedure is the standard surgical treatment of d transposition of great arteries (d-TGA), ventricular septal defect (VSD), and pulmonary stenosis. Late morbidity is significant due to recurrent left ventricular outflow obstruction (LVOTO), early conduit obstruction, and arrhythmias, with troublesome late mortality. To avoid recurrent LVOTO, we routinely enlarge the VSD and resect the infundibular septum before LV baffling to the aorta. We examined the efficacy of this approach in mitigating recurrent LVOTO risk. METHODS: Late echocardiographic and time-related clinical results of patients undergoing the Rastelli procedure were examined. Demographics and operative variables affecting outcomes were analyzed. RESULTS: The Rastelli cohort comprised 36 patients with d-TGA, VSD, and pulmonary stenosis. Median age at operation was 2.4 years (range, 0.3 to 8.3 years). Pulmonary stenosis was present in 31 and atresia in 5. Twenty-two patients had undergone a previous aortopulmonary shunt, and 6 had an atrial septectomy. No operative or late deaths occurred. Time-related freedom from permanent pacemaker implantation, recurrent LVOTO on echocardiogram, and conduit replacement at 10 years was 82%, 100%, and 49%, respectively. Systolic function was normal in all but 3 patients and 92% were in New York Heart Association functional class I and II. None of the patients had late arrhythmias or required heart transplantation. CONCLUSIONS: Early and midterm survival after the Rastelli procedure is satisfactory. Aggressive resection of the infundibular septum to enlarge the VSD has mitigated the risk of LVOTO recurrence. Late conduit obstruction remains an important source of morbidity and frequently requires reintervention. PMID- 19559213 TI - Assessment of the level of sedation in children after cardiac surgery. AB - BACKGROUND: There is no reference method for the evaluation of the level of sedation in children after cardiac surgery. The utility of the bispectral index and middle latency auditory evoked potentials has not been evaluated. METHODS: The bispectral index, middle latency auditory evoked potentials, Ramsay scale, and COMFORT scale were used for assessment of the level of sedation in critically ill children after cardiac surgery and other surgical procedures. The measurements with these four methods were recorded simultaneously once a day for five days. The level of sedation was categorized in two levels, moderate or deep, according to the values obtained from each method. Correlations and agreements among the methods and the best bispectral index and middle latency auditory evoked potential values that discriminated between the two levels of sedation were calculated. RESULTS: Thirty-two children after cardiac surgery were included in the study, together with eighteen children after other surgical procedures who formed the control group. In each group, the correlation and agreement between the four methods varied between moderate and good. In the cardiac surgery patients, when the level of sedation was determined by the Ramsay scale, the best values of bispectral index and middle latency auditory evoked potentials that discriminated between the two levels of sedation were 63.5 and 37.5, respectively, and these values predicted the level of sedation correctly in 84.4% of the patients with each method. CONCLUSIONS: Bispectral index and middle latency auditory evoked potentials could be useful to assess the level of sedation in children after cardiac surgery. PMID- 19559214 TI - Invited commentary. PMID- 19559216 TI - Invited commentary. PMID- 19559215 TI - Interinstitutional comparison of risk-adjusted mortality and length of stay in congenital heart surgery. AB - BACKGROUND: Risk Adjustment for Congenital Heart Surgery (RACHS) and basic Aristotle scores (BCS) have been shown to correlate with mortality and length of stay (LOS) after congenital heart surgery. Interinstitutional comparisons using these scores, as well as comprehensive Aristotle score (CCS), have not been demonstrated. METHODS: We recorded age, weight, RACHS, BCS, CCS, mortality, and LOS for 1,103 patients undergoing cardiac surgery between September 1, 2004, and June 1, 2007, at two institutions. We used binary logistic and multiple linear regressions to evaluate determinants of mortality and LOS, respectively, the C statistic to compare the predictive power of the three scoring systems for mortality, the odds ratio to compare the two institutions, and regression coefficients to compare scoring systems and institutions for LOS. RESULTS: Raw mortality was 2.9% at both institutions. Final logistic regression models contained only CCS. Odds ratios for death at institutions 1 and 2 were 1.25 and 1.26, respectively (not significant). C statistics for RACHS, BCS, and CCS were 0.73, 0.63, and 0.81, respectively (p = 0.01 for CCS versus BCS; p = 0.02 for CCS versus RACHS). Final regression model for LOS retained age, RACHS, and CCS (R(2) = 0.44). The RACHS regression coefficient was greater for institution 2. CONCLUSIONS: The CCS tends to have more predictive power than RACHS and BCS for mortality. The LOS is moderately correlated with CCS, RACHS, and age together, but the model is a poor predictor of individual LOS. The LOS for RACHS category 6 cases differed between the institutions. This study suggests methods that can be used to compare institutions in a risk-adjusted manner. PMID- 19559217 TI - One thousand repeat sternotomies for congenital cardiac surgery: risk factors for reentry injury. AB - BACKGROUND: Reentry injury is a risk associated with repeat sternotomy for cardiac surgery. This risk has been well defined for adults, but there is less information available for patients with congenital heart disease. The goal of this review was to identify the incidence, risk factors, and outcomes for reentry injury in patients with congenital heart disease. METHODS: Eight hundred two patients with congenital heart disease had 1,000 consecutive repeat sternotomies between August 2000 and November 2007. Records were reviewed for demographics, history, operative techniques, and outcomes. Univariate risk factors for reentry injury and operative mortality were assessed. RESULTS: Median age and weight were 2.1 years (range, 0.1 to 34.6 years) and 11 kg (range, 2.5 to 123 kg). There were 639 second, 287 third, and 74 fourth or higher sternotomies. There were 13 reentry injuries (1.3%) involving right ventricle-pulmonary artery conduits (n = 4), aorta or aortic conduits (n = 3), right ventricular outflow tract patches or pseudoaneurysms (n = 3), and others (n = 3). Risk factors for injury were presence of a right ventricle-pulmonary artery conduit (6 of 115 with conduit [5.2%] versus 7 of 885 without [0.8%]; p < 0.001) and sternotomy number (relative risk, 2.28; p < 0.001). Reentry injury was associated with longer procedure times (median, 420 minutes with injury versus 248 without; p < 0.001). Operative mortality occurred in 18 patients and was associated with sternotomy number and procedure time (p < 0.001), but not reentry injury (p = 0.2). CONCLUSIONS: Risk of reentry injury during repeat sternotomy for congenital heart disease is low. Increasing sternotomy number and the presence of a right ventricle-pulmonary artery conduit are risk factors for reentry injury. However, reentry injury is not associated with increased risk of operative mortality. PMID- 19559218 TI - Selective cerebral perfusion: real-time evidence of brain oxygen and energy metabolism preservation. AB - BACKGROUND: Deep hypothermic circulatory arrest (DHCA) is commonly used for complex cardiac operations in children, often with selective cerebral perfusion (SCP). Little data exist concerning the real-time effects of DHCA with or without SCP on cerebral metabolism. Our objective was to better define these effects, focusing on brain oxygenation and energy metabolism. METHODS: Piglets undergoing cardiopulmonary bypass were assigned to either 60 minutes of DHCA at 18 degrees C (n = 9) or DHCA with SCP at 18 degrees C (n = 8), using pH-stat management. SCP was administered at 10 mL/kg/min. A cerebral microdialysis catheter was implanted into the cortex for monitoring of cellular ischemia and energy stores. Cerebral oxygen tension and intracranial pressure also were monitored. After DHCA with or without SCP, animals were recovered for 4 hours off cardiopulmonary bypass. RESULTS: With SCP, brain oxygen tension was preserved in contrast to DHCA alone (p < 0.01). Deep hypothermic circulatory arrest was associated with marked elevations of lactate (p < 0.01), glycerol (p < 0.01), and the lactate to pyruvate ratio (p < 0.001), as well as profound depletion of the energy substrates glucose (p < 0.001) and pyruvate (p < 0.001). These changes persisted well into recovery. With SCP, no significant cerebral microdialysis changes were observed. A strong correlation was demonstrated between cerebral oxygen levels and cerebral microdialysis markers (p < 0.001). CONCLUSIONS: Selective cerebral perfusion preserves cerebral oxygenation and attenuates derangements in cerebral metabolism associated with DHCA. Cerebral microdialysis provides real-time metabolic feedback that correlates with changes in brain tissue oxygenation. This model enables further study and refinement of strategies aiming to limit brain injury in children requiring complex cardiac operations. PMID- 19559219 TI - An artificial right ventricle for failing fontan: in vitro and computational study. AB - BACKGROUND: The aim of this study is to develop a destination low-pressure artificial right ventricle (ARV) to correct the impaired hemodynamics in the failing Fontan circulation. METHODS: An in vitro model circuit of the Fontan circulation was created to reproduce the hemodynamics of the failing Fontan and test ARV performance under various central venous pressures (CVP) and flows. A novel geometry of the extracardiac conduit was designed to adapt to the need of the pump. The ARV was a low-pressure axial flow pump designed to produce a low suction inflow pressure and moderate outflow increase. With the power off, the passive forward gradient across the propeller is 2 mm Hg at 4.5 L/min. The ARV would require 4 watts at a rotation of 5000 rpm. To examine the shear loading on the red blood cells, virtual particles were injected upstream of the ARV inducer and tracked by computerized modeling. RESULTS: The effect of the ARV on the failing Fontan was studied at various CVP pressures and flows, and under constant values of lung resistances and left atrial pressure set respectively to 2.5 Woods Units and 7 mm Hg. The CVP pressures decreased respectively from 25, 22.5, 20, 17.5, 15, and 10 mm Hg to a minimal value of 2 to 5 mm Hg with a pump speed varying from 1700 to 4500 rpm. The pulmonary artery pressures increased moderately between 12.5 and 25 mm Hg at 4500 rpm. Cardiac output at 4500 rpm was increased by an average gain of 2 L/min. The average blood damage index was 0.92%, far below the 5% value considered to cause hemolysis. The flow structure produced by the pump was suitable. CONCLUSIONS: The performance of this novel low pressure ARV was satisfactory, showing good decrease of CVP pressures, a moderate increase of pulmonary artery pressures, adequate increase of cardiac output, and minimal hemolysis. The use of a mock Fontan model circuit facilitates device prototyping and design to a far greater extent than can be achieved using animal studies, and is an essential first step for rapid design iteration of a novel ARV device. The next steps are the manufacturing of this device, including an electromagnetic engine, a regulatory system, and further testing the device in a survival animal experiment. PMID- 19559220 TI - Invited commentary. PMID- 19559221 TI - Analysis of cervical esophagogastric anastomotic leaks after transhiatal esophagectomy: risk factors, presentation, and detection. AB - BACKGROUND: Transhiatal esophagectomy with cervical esophagogastric anastomosis is a common approach in patients requiring esophagectomy. Factors for developing cervical esophagogastric anastomosis leaks (CEGAL), their presentation, and the value of a routine postoperative screening barium swallow in detecting CEGALs and other complications were analyzed. METHODS: This single-institution retrospective study used medical records and an esophagectomy database to assess results in 1,133 patients who underwent transhiatal esophagectomy and a cervical esophagogastric anastomosis, 241 for benign disease and 892 for cancer, between January 1996 and December 2006. RESULTS: Esophagectomy patients who experienced CEGALs included 127 (14.2%) with cancer and 23 (9.5%) with benign disease. Logistic regression analysis identified increasing number of preoperative comorbidities (p < 0.001), active smoking history (p = 0.044), and postoperative arrhythmia (p = 0.002) as risk factors for CEGALs, and a side-to-side stapled cervical esophagogastric anastomosis compared with a manually sewn one as protective (p < 0.001). For cancer patients, higher pathologic stage disease (p = 0.050) was a risk factor for CEGALs. For patients with benign disease, a higher number of prior esophagogastric operations (p = 0.007) is a risk factor for CEGALs. Of the 90.7% of CEGALs that occurred on or before postoperative day 10, cervical wound drainage (63.3%) was the most common presenting symptom. Screening barium swallow identified postoperative complications and influenced outcome in 39 patients (3.8%). CONCLUSIONS: Higher number of preoperative comorbidities, advanced pathologic stage, postoperative arrhythmia, an increased number of prior esophagogastric surgeries, and active smoking history are risk factors for developing CEGAL, and a side-to-side stapled cervical esophagogastric anastomosis is protective. Screening barium swallow identifies few postoperative complications, but provides quality control. PMID- 19559222 TI - Predictors of long-term survival after resection of esophageal carcinoma with nonregional nodal metastases. AB - BACKGROUND: Patients with esophageal carcinoma and celiac, cervical, or other nonregional nodal metastases generally have a poor prognosis after surgical resection. Factors predicting long-term survival are unclear. The goal of this study was to analyze factors predicting long-term survival in this subset of patients. METHODS: We conducted a retrospective review of a prospective database over a 20-year period to identify patients with resected esophageal carcinoma with nonregional lymph node metastases. Medical records were reviewed and risk factors were analyzed. RESULTS: Sixty-seven patients underwent esophagectomy for M1a or M1b disease from 1987 to 2007. Esophagectomy was transthoracic in 62 patients and transhiatal in 5. The median number of lymph nodes harvested was 36. Sites of nodal metastases were the following: recurrent nodal chain in 42 patients, celiac in 20, both recurrent and celiac in 4, and paratracheal in 1. Median length of follow-up was 66 months. The 5-year overall survival for the entire cohort was 25%. The 5-year overall survival was significantly higher with earlier T-status, (pathologic tumor [pT]1/T2 vs pT3/T4; 62% vs 15%, p = 0.006). Thirteen patients who had nonregional nodal metastases without involvement of regional nodes (pN0) had a significant improvement in 5-year survival (67% vs 15%; p < 0.001). Patients with squamous cell carcinomas had higher 5-year survival compared with those with adenocarcinomas (42% vs 14%; p = 0.009). Patients treated with induction chemotherapy had prolonged 5-year survival (41%, p = 0.06) compared with those treated with adjuvant chemotherapy (11%) or no therapy (20%). Multivariate analysis demonstrated that chemotherapy treatment, squamous cell type, and early T stage (pT1/T2) are significant positive predictors of survival. CONCLUSIONS: Surgical resection for patients with esophageal cancer associated with nonregional nodal metastases results in 25% survival at five years. Squamous histology, earlier T status, and perioperative chemotherapy are independent positive predictors of long-term survival. PMID- 19559223 TI - Esophageal stent placement for the treatment of spontaneous esophageal perforations. AB - BACKGROUND: Traditional therapy for spontaneous esophageal perforation has most often been urgent operative repair. This investigation summarizes the treatment of spontaneous perforations of the esophagus using an occlusive removable esophageal stent. METHODS: During a 48-month period, patients with a spontaneous esophageal perforation were offered endoluminal esophageal stent placement as the initial therapy instead of operation. Excluded were patients with an esophageal malignancy or a chronic esophageal fistula. Silicone-coated stents were placed endoscopically using general anesthesia and fluoroscopy. Adequate drainage of infected areas was achieved. Leak occlusion was confirmed by esophagram. RESULTS: Twenty-one esophageal stents were placed in 19 patients for spontaneous esophageal perforations. Associated endoscopic (n = 19) or surgical procedures (n = 9) were also simultaneously performed. Leak occlusion occurred in 17 patients (89%). Fifteen patients (79%) were able to initiate oral nutrition within 72 hours of stent placement. Two patients (10%) with a perforation extending across the gastroesophageal junction experienced a continued leak after stent placement and underwent operative repair. Stent migration in 4 patients (21%) required repositioning (n = 4) or replacement (n = 2). Stents were removed at a mean of 20 +/- 15 days after placement. Hospital length of stay was 9 +/- 12 days. CONCLUSIONS: Endoluminal esophageal stent placement is an effective treatment of most spontaneous esophageal perforations. These stents result in rapid leak occlusion, provide the opportunity for early oral nutrition, may significantly reduce hospital length of stay, are removable, and avoid the potential morbidities of operative repair. PMID- 19559224 TI - Invited commentary. PMID- 19559225 TI - Intrathoracic lymph node metastases from extrathoracic carcinoma: the place for surgery. AB - BACKGROUND: Intrathoracic hilar or mediastinal lymph node metastases (HMLNMs) of extrathoracic carcinomas are infrequent. Their treatment strategy is not established and their prognosis poorly known. We reviewed the place of surgical intervention in their management. METHODS: Among 565 patients with mediastinal lymph node enlargement, 37 had a history of extrathoracic carcinoma. The enlargement consisted in HMLNMs in 26 (15 men, 11 women), with a mean age of 57.6 (range 19-78) years. Surgical procedures were reviewed. RESULTS: Diagnostic procedures, comprising mediastinoscopy in 9, anterior mediastinotomy in 2, and video-assisted thoracic surgery (VATS) in 4, were performed mainly because of unresectability due to diffuse and bilateral HMLNMs. Cancer location was breast in 6, kidney or prostate in 2 each, and bladder, rectum, testis, melanoma, and larynx in 1 each. Median survival was 21 months. Resection was performed in 11 patients, comprising posterolateral thoracotomy in 6, muscle sparing thoracotomy in 2, and VATS in 3. Seventeen involved LN stations were removed; of these, primary were kidney in 3, testis or thyroid in 2 each, and larynx, nasopharynx, and intestinum in 1 each. Five-year survival was 41.6% (median, 45 months). CONCLUSIONS: HMLNMs of extrathoracic carcinoma may be isolated, probably in the context of a particular lymphatic mode of spread. Our experience demonstrates that operation is mainly diagnostic but resection may safely achieve local control of the disease and deserves being advocated in patients with isolated and resectable HMLNMs. PMID- 19559226 TI - Invited commentary. PMID- 19559228 TI - Transdiaphragmatic harvesting of the omentum through thoracotomy for bronchial stump reinforcement. AB - BACKGROUND: We present our technique of omental flap transposition performed through a thoracotomy for bronchial stump protection, and employed over 11 years. METHODS: Between February 1997 and January 2008, the transdiaphragmatic harvesting of the omentum was performed, using an original technique through a thoracotomy approach, in 45 patients. Forty-three patients (29 male, 14 female), considered at high risk for bronchial dehiscence, simultaneously underwent pneumonectomy and 2 patients (1 male, 1 female) were treated for an early postpneumonectomy bronchopleural fistula by the standard thoracotomy route. The omental flap was mobilized through a radial incision in the diaphragm avoiding an additional laparotomy. The only contraindication for this technique was a previous abdominal intervention. Duration of follow-up ranged between 6 and 102 months (median, 46). RESULTS: There were no complications related to the omentoplasty. Major complications related to pneumonectomy occurred in 4 patients (9%). Perioperative mortality rate was 2.1% (1 of 45). The non-life threatening complication rate was 11.1% (5 of 45). Postoperative hospital stay ranged between 5 and 21 days (median, 8.3) in the 43 patients undergoing prophylactic omentoplasty and was 11 and 14 days, respectively, in the 2 patients receiving omentoplasty after bronchial dehiscence. No neoplastic recurrence on the bronchial stump or late fistula occurred during follow-up. CONCLUSIONS: This technique of omental flap transposition for bronchial stump coverage through a thoracotomy is an effective method for the prevention and treatment of postpneumonectomy bronchopleural fistula. The amount of omentum obtained by this technique is appropriate for bronchial reinforcement but not for filling the pleural cavity. This procedure can be performed safely through thoracotomy access avoiding an additional laparotomy. PMID- 19559227 TI - Surgical resection of pulmonary malignant tumors after living donor liver transplantation. AB - BACKGROUND: The aim of this study is to report on patients who developed tumor recurrence of the lung or de novo pulmonary malignancies after living donor liver transplantation (LDLT) and to show the benefit of a surgical resection for these pulmonary malignant tumors. METHODS: A total 246 patients who underwent LDLT were investigated. RESULTS: Pulmonary malignant tumors after LDLT were observed in 12 (4.9%) of 246 patients studied. These patients included 9 tumor recurrences and 3 de novo malignancies. The frequency of pulmonary recurrence was 9.4% (9 of 96 patients) and that of pulmonary de novo malignancies including 2 primary lung cancer and 1 mucosa-associated lymphoid tissue (MALT) lymphoma, was 1.2% (3 of 246 patients). Four of 9 recurrent patients could undergo surgical resections and the survival range in patients who received surgery was 17 to 56 months with a mean of 36 months after LDLT; on the other hand, the survival range in patients that could not undergo a surgical resection was 4 to 26 months with a mean of 18 months. Among the de novo malignancies, only the MALT lymphoma patient could undergo a surgical resection. Repeated surgical resections of pulmonary malignant tumors could be performed in 3 patients and all these patients have been long term survivors. CONCLUSIONS: These results suggest a surgical resection of pulmonary malignancies including tumor recurrences or de novo malignancies after LDLT is a feasible procedure and may prolong survival in selected patients, even under immunosuppressive conditions. PMID- 19559229 TI - The safe transition from open to thoracoscopic lobectomy: a 5-year experience. AB - BACKGROUND: We hypothesized that established thoracic surgeons without formal minimally invasive training can learn thoracoscopic lobectomy without compromising patient safety or outcome. METHODS: Data were retrospectively collected on patients who underwent pulmonary lobectomy at a single health system between August 1, 2003, and April 1, 2008. Age, sex, pulmonary function tests, preoperative and postoperative stages, pathologic diagnosis, anatomic resection, extent of lymph node sampling, surgical technique and duration, complications, blood loss, transfusion requirement, chest tube duration, length of hospital stay, 30-day readmission, and mortality rate were examined. The percentage of patients who underwent thoracoscopic lobectomy and their outcomes were then compared among three chronologic cohorts. RESULTS: Three hundred sixty-four patients underwent pulmonary lobectomy (239 open; 99 thoracoscopic; 26 thoracoscopic converted to open). Baseline characteristics, staging, pathologic diagnosis, and anatomic resections were similar in the early, middle, and late cohorts. The percentage of thoracoscopic lobectomies increased from 16% to 49%, whereas open lobectomy decreased from 81% to 42% (p < 0.0001). The complication rate remained constant with the exception of air leaks lasting more than 7 days (9% versus 10% versus 2%; p = 0.02). Hospital length of stay (6 versus 5 versus 4 days; p < 0.0001) and chest tube duration (4 versus 3 versus 3 days; p < 0.0001) decreased and operative duration increased as more thoracoscopic lobectomies were performed. Blood loss, transfusion requirement, 30-day readmission, and 1-year survival were not significantly different among chronologic cohorts. CONCLUSIONS: Established thoracic surgeons can safely incorporate thoracoscopic lobectomy with no increase in morbidity or mortality. PMID- 19559230 TI - Diagnostic surgical lung biopsies for suspected interstitial lung diseases: a retrospective study. AB - BACKGROUND: Current guidelines for interstitial lung disease support a surgical biopsy for optimal diagnosis and treatment, yet only a minority of patients undergo such biopsy. Our objectives were to address the properties of a surgical lung biopsy for suspected interstitial lung disease, the diagnostic yield of the procedure, and whether it resulted in changes in diagnosis and treatment. METHODS: A retrospective nationwide study including 73 patients (mean age, 57.3 years; 58% males) who underwent a surgical lung biopsy for suspected interstitial disease in Iceland between 1986 and 2007 was conducted. Patient records and histologic specimens were reviewed. Before the surgical biopsy a transbronchial or computed tomography-guided biopsy had been performed in two thirds of the patients. RESULTS: The complication rate for surgical lung biopsy was 16%, and 30 day operative mortality was 2.7%, both significantly higher in patients with preoperative respiratory failure. After the procedure, a definite histopathologic diagnosis was obtained in 81% of the patients. Usual interstitial pneumonia was the most common diagnosis (31%). The clinical diagnosis was changed for 73% of the patients, and in 53% of the patients the biopsy resulted in changes in treatment. CONCLUSIONS: Surgical lung biopsy is a powerful tool for diagnosis of suspected interstitial lung disease. It results in a specific diagnosis for the majority of patients and changes in treatment for more than half. Operative morbidity and mortality are low but still significant, so patients should be carefully selected for the procedure, especially those with respiratory failure. PMID- 19559231 TI - Invited commentary. PMID- 19559232 TI - Operative strategies for pulmonary artery occlusion secondary to mediastinal fibrosis. AB - BACKGROUND: Fibrosing mediastinitis is a rare disease characterized by an excessive fibrotic reaction in the mediastinum, which may entrap mediastinal structures including the pulmonary arteries. Our objectives were to assess the surgical strategies and outcomes of repair of pulmonary artery occlusion attributable to mediastinal fibrosis. METHODS: With approval from the Mayo Clinic Institutional Review Board, we identified all patients with fibrosing mediastinitis who underwent an operation for relief of pulmonary artery obstruction between 1980 and 2008. Perioperative data were collected using medical records and late follow-up surveys. RESULTS: Operative procedures to bypass or reconstruct an obstructed pulmonary artery were performed in 5 patients. Patients' median age was 40 years (range, 27 to 51 years), and all patients were symptomatic and had right ventricular hypertension. In 3 patients, a double-outlet right ventricle was constructed using a valved conduit (porcine valved conduit, n = 1; aortic homograft, n = 2) from the right ventricle to the right pulmonary artery. Two patients required complete reconstruction of the pulmonary artery confluence using a pulmonary homograft in 1 patient and a hybrid technique of autologous pericardial reconstruction and intraoperative stenting in another patient. All patients had a reduction in right ventricular pressures after operation. One patient died perioperatively owing to respiratory failure; the remaining 4 patients were alive at a median follow-up of 7.4 years (range, 0.5 to 14.7 years). One patient required late balloon dilatation of the conduit and distal pulmonary arteries 10 years after initial operation, but the remaining conduits were widely patent at late follow-up. Late functional improvement was limited owing to other complications from mediastinal fibrosis or other comorbidities. CONCLUSIONS: Treatment of pulmonary artery occlusion attributable to mediastinal fibrosis can be challenging. Successful operative strategies include both creation of a double-outlet right ventricle and complete reconstruction of the pulmonary artery confluence. Hybrid techniques of both conduit placement and stenting should also be considered for patients with occluded pulmonary arteries. PMID- 19559233 TI - Bilateral thoracoscopic T2 to T3 sympathectomy versus botulinum injection in palmar hyperhidrosis. AB - BACKGROUND: Bilateral T2 to T3 thoracoscopic sympathectomy and injection of botulinum toxin-A are presently the most effective modalities in the treatment of primary palmar hyperhidrosis. In this study we evaluated comparative merits of the two therapies. METHODS: Patients suffering primary palmar hyperhidrosis were treated by either bilateral T2 to T3 thoracoscopic sympathectomy (n = 68) or by injection of botulinum toxin-A (n = 86). The groups were homogeneous for relevant demographic, physiologic, and clinical data. Quantification of sweat production was performed by Minor's iodine starch and glove tests. Subjective changes were assessed by quality of life questionnaires (Hyperhidrosis, Dermatology Life Quality Index, Short Form-36, Nottingham's Health Profile) and patient's satisfaction self-assessment. A cost comparison between groups was also carried out. RESULTS: No operative mortality or major morbidity was recorded in either group. Minor's test showed a more significant reduction in the surgical group: +94% versus +63% at 6 months and +94% versus +30% at 12 months. Compensatory sweating was significantly greater and long-lasting in the surgical group. All subjective tests improved rapidly and significantly in both groups. After 6 months, results mildly worsened in the surgical group and more significantly in the botulinum group. Patient's satisfaction was initially greater in the botulinum group (p = 0.03), but after 6 months it significantly reversed (p = 0.04). Surgical treatment cost approximately as much as four botulinum treatments. CONCLUSIONS: Thoracoscopic sympathectomy is superior to botulinum toxin-A injection. The greater initial costs and discomfort are offset by a greater reduction in compensatory sweating. PMID- 19559234 TI - Thoracic duct ligation for persistent chylothorax after pediatric cardiothoracic surgery. AB - BACKGROUND: There is considerable literature on incidence and medical management of postsurgical chylothorax in children but little is known about outcomes of thoracic duct ligation (TDL) for patients refractory to medical therapy. METHODS: A retrospective review of patients undergoing TDL after cardiothoracic surgery (1992 through 2007) was done. Data on demographics including cardiac morphology, characteristics of chylous drainage, medical management, and post-TDL course were collected. When available, imaging studies of the upper body venous drainage vessels were examined. RESULTS: Twenty patients (median age, 0.65 years; range, 0.03 to 11 years; weight, 7.0 kg; range, 2.6 to 30 kg) had a diagnosis of chylothorax made 8.5 days (range, 2 to 118 days) after initial operation. Median duration of pre-TDL medical management was 17.5 days (range, 7 to 69 days). Median drainage for 5 days preceding TDL was 34.5 mL x kg(-1) x d(-1) (range, 15 to 135 mL x kg(-1) x d(-1)) with maximal output of 65 mL x kg(-1) x d(-1) (range, 30 to 200 mL x kg(-1) x d(-1)). After TDL, there was a decrease in median drainage to 13 mL x kg(-1) x d(-1) (range, 4 to 160 mL x kg(-1) x d(-1); p = 0.003). Chest tubes were removed 8.5 days (range, 4 to 34 days) after TDL. There were 4 deaths (none attributed to TDL), 2 treatment failures (post-TDL chest tube drainage > 2 mL x kg(-1) x d(-1) > 14 days), and 2 recurrences (after initial chylothorax resolution and hospital discharge). Three patients had documented upper body venous thrombosis. Univariate analysis demonstrated thrombosis of upper body venous vessels (p = 0.02) and prolonged post-TDL chest tube drainage (p = 0.01) were risk factors for death, treatment failure, or chylothorax recurrence. CONCLUSIONS: Thoracic duct ligation leads to a major reduction in chest tube drainage and prompt tube removal in most pediatric patients and should be considered early in refractory postoperative chylothorax. Patients with upper body venous thrombosis associated with chylothorax are at a high risk for failure of TDL and mortality. PMID- 19559235 TI - The EmBlocker: efficiency of a new ultrasonic embolic protection device adjunctive to heart valve surgery. AB - PURPOSE: Perioperative cerebral microemboli in cardiac surgery are associated with postoperative neurologic complications. The EmBlocker (Neurosonix Ltd, Rehovot, Israel), a newly developed device should be positioned against the ascending aorta, and it produces an ultrasonic force expected to divert microemboli away from the cerebral vasculature and reduce cerebral emboli. DESCRIPTION: Twenty-one consecutive patients, undergoing a valve procedure, were enrolled into this nonrandomized pilot study. The EmBlocker (Neurosonix Ltd) was positioned in 11 consecutive patients and activated for 1 minute (1.5 W/cm(2)) during seven selected aortic manipulations and for 10 minutes (0.5 W/cm(2)) intermittently after cross-clamp removal. Transcranial Doppler-based quantification of microembolic signals was performed in all patients. EVALUATION: The use of the EmBlocker showed a significant overall reduction of the cerebral microembolic signals of 53%. CONCLUSIONS: The use of the EmBlocker during valve surgeries is associated with a reduction of perioperative cerebral microembolic signals. This new technology holds the potential to lower the risk of postoperative neurologic complications. PMID- 19559236 TI - An experimental study of type I endoleak repair with a suturing device. AB - PURPOSE: An experimental study was done to investigate repair of type I endoleaks in thoracic aortic aneurysms using the T-Fix suturing device (Smith & Nephew Co, Ltd, London, United Kingdom). DESCRIPTION: A saccular descending aortic aneurysm was made in 5 pigs experimentally. A stent graft was deployed to produce a proximal type I endoleak. Under fluoroscopy, the aorta was punctured with the spinal needle with the T-Fix plastic bar, and the plastic bar was deployed with a push rod. A sufficient number of T-Fix sutures were used until angiography revealed that the type I endoleak had disappeared. EVALUATION: No hemodynamic events occurred during the procedure. An average of 2.5 +/- 0.6 T-Fix sutures were required to eliminate the endoleak. The experimental T-Fix repair was performed without any complications. A new method of repairing type I endoleaks for thoracic aortic aneurysms was successfully performed using the T-Fix system. CONCLUSIONS: Although the T-Fix repair currently has some anatomic and clinical limitations, improvement of the device should lead to the increased use of this repair. PMID- 19559237 TI - Iliac arterial intussusception from an aortic endoclamp catheter. AB - Minimally invasive cardiac surgical procedures are gaining widespread acceptance with the advent and development of the femoral route for cardiopulmonary bypass. Aortic endoclamps are widely used and are one of the most important parts of these surgical techniques. This report presents iliac arterial intussusception from an aortic endoclamp catheter, which is a very rare complication with this type of device. Preventative strategies are presented. PMID- 19559238 TI - Subclinical thrombosis of the ascending aorta: a possible paraneoplastic syndrome. AB - Thrombosis of the ascending aorta is a rare, potentially lethal complication. We report the case of a 56-year-old woman with a massive but subclinic thrombosis of the ascending aorta after two cycles of chemotherapy due to an epidermoid lung carcinoma stage T3 N2 M0. An emergent aortic thrombectomy was performed under deep hypothermic circulatory arrest. This thrombotic event occurred in an arterial vessel with high laminar flow, which is extremely uncommon and did not present any clinical manifestation. PMID- 19559239 TI - A modified technique for preventing spinal cord ischemia during type II thoracoabdominal aneurysm repair. AB - A 51-year-old man required replacement of the thoracoabdominal aorta due to a type II thoracoabdominal aortic aneurysm. We tailored and plicated the aortic aneurysm to make a closed tube. All of the intercostal arteries and lumbar arteries were reimplanted using a closed tube constructed with an aneurysmoplasty to the main aortic graft, using this tube to protect the spinal cord. The closed tube maintained blood flow to the intercostal and lumbar arteries, and no neurologic deficits developed. PMID- 19559240 TI - Aortic valve vegetation without endocarditis. AB - We present a 30-year-old man with an acute middle cerebral artery territory infarction. A transesophageal echocardiogram showed a large, highly mobile mass attached to the patient's aortic valve. We discuss the differential diagnosis of a cardiac mass that includes infection, tumor, and thrombus. A complete workup showed no evidence of systemic infection but did reveal the presence of antiphospholipid antibodies. The patient also had a history of a right lower extremity deep venous thrombosis. Anticoagulation therapy was started, and follow up showed complete resolution of the aortic valve lesion. This case highlights that when a valvular vegetation is encountered in a clinical setting that does not suggest infectious endocarditis, the diagnosis of antiphospholipid antibody syndrome should be considered. This case and our review of the literature suggest that vegetations in antiphospholipid antibody syndrome, no matter how large and ominous in appearance, can be treated successfully with anticoagulation and vigilant observation. PMID- 19559241 TI - Mitral valve repair by leaflet sliding and annular downsizing in active infective endocarditis. AB - We repaired a large defect in the posterior mitral leaflet after an extensive removal of infected tissue, using an extended leaflet sliding and annular downsizing with a small prosthetic ring in 2 patients with active endocarditis. PMID- 19559242 TI - Successful treatment of heart failure due to acute transplant rejection with the Impella LP 5.0. AB - Cardiogenic shock resulting from transplant rejection is a serious complication with high mortality and morbidity. Often resistant to maximal medical therapy, this condition frequently requires mechanical circulatory support until recovery or retransplantation. We present a 52-year-old patient with multiorgan failure secondary to acute graft rejection after orthotopic heart transplantation. Maximal medical therapy was not successful, and the patient was bridged to recovery with an Impella LP 5.0 (Abiomed Inc, Danvers, MA) left ventricular assist device (LVAD). The relative merits of this therapeutic approach are outlined and discussed. The patient was discharged 3 weeks after LVAD removal and remains clinically stable. PMID- 19559243 TI - Idiopathic pulmonary artery aneurysm treated with surgical correction and concomitant coronary artery bypass grafting. AB - Idiopathic pulmonary artery aneurysm is a rare clinical entity, and therefore the natural course and clinical management are not well established. We present the case of an elderly woman with a symptomatic idiopathic pulmonary artery aneurysm who underwent surgical repair along with simultaneous coronary artery bypass grafting. With long-term follow-up presented in this report, we describe the safety and durability of surgical repair. PMID- 19559244 TI - Surgical repair of anomalous origin of the left coronary artery arising from the left pulmonary artery. AB - Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital anomaly. We report an extremely uncommon variety of ALCAPA, in which the left coronary artery arose from the left pulmonary artery in a patient who presented with severe heart failure in early infancy. After direct reimplantation of the left coronary artery into the ascending aorta, the patient's cardiac function recovered successfully. PMID- 19559246 TI - Surgical treatment of cardiac pheochromocytoma: a case report. AB - Primary cardiac pheochromocytoma is an extremely rare neoplasm. We report a 15 year-old girl who was presented with paroxysmal hypertension. An iodine-131 metaiodobenzylguanidine scintigraphy scanning showed a pheochromocytoma in her right atrial and ventricular wall. The tumor was subsequently confirmed by magnetic resonance imaging and coronary angiogram. This patient underwent a successful surgical resection of the tumor, a reconstruction of the atrial ventricular wall and right coronary artery bypass grafting. Her blood pressure remained normal thereafter. A follow-up coronary angiogram revealed a patent saphenous vein graft 4 months after the operation. PMID- 19559245 TI - Hybrid treatment of superior vena cava syndrome in a child. AB - A 10-year-old boy with a history of renal failure and hemodialysis by indwelling superior vena cava (SVC) catheters was diagnosed with SVC obstruction and clinically severe SVC syndrome. During attempted recanalization of the SVC in the cardiac catheterization laboratory, he suffered a perforation of his SVC with pericardial tamponade. After treatment of the perforation and relief of tamponade, he underwent a hybrid procedure to recanalize his SVC. A needle and then guidewire were passed directly from the right atrium through the SVC obstruction and were used to successfully dilate and stent the obstruction. PMID- 19559247 TI - Idiopathic pulmonary vein thrombosis: a rare cause of massive hemoptysis. AB - The case history of an adult female with massive hemoptysis due to idiopathic left inferior pulmonary vein thrombosis necessitating lower lobectomy is presented with a review of the current literature. PMID- 19559248 TI - Inadvertent total spinal anesthesia after intercostal nerve block placement during lung resection. AB - Intercostal nerve block is a recognized way of providing analgesia at thoracotomy. There is a rare association between intercostal nerve block and the complication of total spinal anesthesia. This may arise inadvertently by injection into a dural cuff extending outside the intervertebral foramen. We report our experience with a patient who sustained this life-threatening complication. The patient required postoperative ventilation until the neurologic deficits resolved. The operator must be aware that intercostal nerve block runs the rare but potentially fatal risk of total spinal block. PMID- 19559249 TI - Managing pulmonary artery catheter-induced pulmonary hemorrhage by bronchial occlusion. AB - A 76-year-old woman underwent mitral valve repair and coronary artery bypass grafting. Intrabronchial bleeding occurred after inflation of the balloon tip of the pulmonary artery catheter in the wedge position. A Forgaty catheter was introduced into the trachea parallel to the endotracheal tube and advanced under bronchoscopic vision into the intermediate bronchus. Tamponade of the bleeding was achieved by by filling the Forgaty balloon tip with saline. Weaning from extracorporeal circulation was uneventful. On the first postoperative day, the Forgaty catheter was removed and bronchial lavage of the middle and lower lobe was performed without any additional bleeding complication. PMID- 19559250 TI - Severe hypoxemia due to intrapulmonary shunting requiring surgery for bronchioloalveolar carcinoma. AB - Bronchioloalveolar carcinoma is a rare, but well-known disease that symptomatically worsens with intrapulmonary shunting and consequent hypoxemia. Surgical resection of the involved area offers relief from disabling hypoxemia and may improve survival. We present 3 patients with intrapulmonary shunting. PMID- 19559251 TI - Novel method to repair tracheal defect by pectoralis major myocutaneous flap. AB - Inflammatory myofibroblastic tumor is extremely uncommon in the trachea. Surgery is recommended when airway obstruction becomes evident. The surgical technique and material used for repairing a massive tracheal defeat is a challenge for the thoracic surgeon. We present a case of repair and reconstruction of a massive defect of the thoracic trachea and right mainstem bronchus with a pectoralis major myocutaneous flap after resection of an inflammatory myofibroblastic tumor. The myocutaneous flap provides reliable material to repair and reconstruct a massive central airway defect. This novel surgical procedure may present new strategies for the treatment of extensive defects of the trachea. PMID- 19559252 TI - Extralobar sequestration in anterior mediastinum with pericardial agenesis. AB - We report a very rare case of extralobar sequestration and pericardial agenesis in a 22-year-old man. A computed tomographic (CT) scan demonstrated an anterior mediastinal mass. No aberrant artery was preoperatively identified. The patient underwent surgery with an impression of thymoma. An extralobar sequestration receiving its blood supply from the left pulmonary artery, accompanied with pericardial agenesis, was noted at the time of operation. The anterior mediastinum is an unusual site for extralobar sequestions. It is recommended to include extralobar sequestration in the differential diagnosis of anterior mediastinal masses, even if the aberrant artery is not recognized on the computed tomographic scan. PMID- 19559253 TI - Sclerosing mediastinitis mimicking anterior mediastinal tumor. AB - A 54-year-old asymptomatic man presented with an anterior mediastinal mass discovered on chest roentgenogram. Chest computed tomography revealed a noncalcified round mass in the mediastinum. A white solid mass, 5-cm in diameter, had arisen from the pericardial adipose tissue with multiple small nodular lesions mimicking mediastinal tumor with pleural dissemination. Postoperative pathologic examination confirmed a diagnosis of sclerosing mediastinitis. Details of the clinical and radiographic feature are presented. PMID- 19559254 TI - Limb-threatening ischemia secondary to a congenital acromioclavicular remnant. AB - Upper extremity vascular compromise from thoracic outlet syndrome is rare and is usually the result of a "cervical rib," anterior scalene muscle abnormality, or clavicular trauma. We report a case of acute axillary artery thrombosis secondary to a congenital acromioclavicular remnant in a 40-year-old woman. PMID- 19559255 TI - A case of primary synovial sarcoma of the thorax with a variant SYT-SSX1 fusion transcript. AB - With synovial sarcoma (SS) of the thorax, being exceptionally rare, its definite diagnosis is difficult, and the optimal therapy has not yet been established. An examination of our patient, a 64-year-old man with SS using a chest roentgenogram showed a large mass with homogeneous density in the lower two-thirds of the left hemithorax. A computed tomographic image of the chest revealed a large, heterogeneous, enhanced mass in the left hemithorax. Histologic examination of the resected tumor tissues suggested monophasic fibrous SS. A fragment of the SYT SSX1 fusion transcript, which was smaller than the control, was amplified with reverse transcriptase polymerase chain reaction. Direct sequence analyses revealed the fusion between exon 9 of SYT and exon 5 of SSX1 instead of fusion between exon 10 of SYT and exon 6 of SSX1, which is found in most cases. Although the biological and clinical significance of this rare variant is not yet known, our data present another example of the usefulness of molecular analyses for making a definite diagnosis of SS in unusual sites. PMID- 19559256 TI - Bronchial angiolipoma. AB - Angiolipoma occurs preferentially in the extremities and trunk. We present a patient with involvement of the bronchus and describe successful localized resection of the lesion. PMID- 19559257 TI - Left circumflex to bronchial artery fistula. PMID- 19559259 TI - An unusual location of a persistent vein of Marshall. PMID- 19559258 TI - Harlequin syndrome. PMID- 19559260 TI - Hemothorax caused by a solitary costal exostosis. PMID- 19559261 TI - Combined open proximal and stent-graft distal repair for distal arch aneurysms: an alternative to total debranching. AB - We present herein a novel, combined, simultaneous open proximal and stent-graft distal repair for complex distal aortic arch aneurysms involving the descending aorta. In the first surgical step, the transverse arch is opened during selective antegrade cerebral perfusion, and a Dacron graft (DuPont, Wilmington, DE) is positioned down the descending aorta in an elephant trunk-like fashion with its proximal free margin sutured circumferentially to the aorta just distal to the left subclavian or left common carotid artery. With the graft serving as the new proximal landing zone, subsequent endovascular repair is performed antegrade during rewarming through the ascending aorta. PMID- 19559263 TI - Ventricular septal defect closure in Taussig-Bing heart: the "pulmonic rule". AB - Accurate ventricular septal defect patch sizing and tailoring remain challenging in many surgical procedures. Surgical exposure frequently limits complete visualization of the ventricular septal defect. Moreover, examination of the heart cavity under cardioplegic arrest may lead to skewed appreciation of the ventricular septal defect caliber and shape. Here we describe a simple and safe surgical tip to predict the size and shape of the ventricular septal defect patch in Taussig-Bing malformation before starting extracorporeal circulation. The patch should be circular with a diameter equal to the under pressure, proximal, pulmonary artery diameter. PMID- 19559262 TI - Preventing blood loss during application of the HEARTSTRING proximal seal system. AB - The HEARTSTRING Proximal Seal System (Guidant Corp, Santa Clara, CA) is used to avoid aortic clamping while the proximal anastomoses are sewn. To protect surgeons from spurting blood while the device is used, we use a see through plastic sheet to cover the area being operated on. This modified technique is applied whenever the system is used and allows the safe use of the device even in high-risk patients with hepatitis or human immunodeficiency virus. PMID- 19559264 TI - A cervical approach to investigating pleural disease. AB - We describe a modern cervical approach to the pleural space using video mediastinoscopy, which allows both mediastinoscopy and pleuroscopy to be performed simultaneously. Mediastinoscopy is carried out with lymph node sampling, and the pleura are exposed and the pleural cavity is entered under direct vision. A thoracoscope is admitted into the pleural space, where lavage, biopsy, and pleurodesis can be carried out. Fifteen patients underwent mediastino pleuroscopy to investigate pleural effusion and stage malignancy. One patient underwent bilateral pleuroscopy through a single cervical approach. There were no mortalities and the mean postoperative stay was 2.4 days. Mediastino-pleuroscopy is safe, uses a small incision, is well tolerated, and allows access to both pleura and the mediastinum. PMID- 19559265 TI - Thrombin in myocardial ischemia-reperfusion during cardiac surgery. AB - Thrombin is a multifunctional protease with procoagulant, pro-inflammatory, and pro-apoptotic effects. Thrombin has direct potentially adverse effects on the endothelium and on cardiomyocytes, which are independent of its procoagulant effects, and it has emerged as a possible mediator of ischemia-reperfusion injury. Several lines of experimental evidence specifically implicate thrombin to be involved in myocardial ischemia-reperfusion injury. Cardiopulmonary bypass increases thrombin generation progressively, but reperfusion after myocardial ischemia induces an additional distinct and rapid increase in thrombin generation. Clinical studies have shown that thrombin formation during cardiac surgery, especially during myocardial reperfusion, is involved with myocardial damage and impaired hemodynamic recovery. Therefore, strategies to improve thrombin control during cardiopulmonary bypass might be beneficial. PMID- 19559266 TI - Postoperative inflammatory reaction and atrial fibrillation: simple correlation or causation? AB - Atrial fibrillation after cardiac operations is a source of morbidity and resource consumption. This systematic review of literature analyzes the current evidence on its pathophysiologic link with the systemic inflammatory response elicited by surgery and cardiopulmonary bypass. Meta-analysis of randomized studies on the effect of off-pump surgery or statin pre-treatment on the incidence of atrial fibrillation was performed. The concept of inflammation as a pathophysiologic determinant of postoperative atrial fibrillation is supported by the literature. The modulation of post-cardiopulmonary bypass systemic inflammation will probably represent a major therapeutic goal in the prevention of postoperative atrial fibrillation. Statins seem to be the most promising pharmacological strategy. PMID- 19559267 TI - Bhagavant Kalke and his pioneering work on the bi-leaflet heart valve prosthesis. PMID- 19559268 TI - Mitral repair is not superior to replacement in elderly patients. PMID- 19559270 TI - Long-term follow-up of the frozen elephant trunk technique for distal aortic arch aneurysm. PMID- 19559271 TI - Postoperative delirium in cardiac operations: microembolic load is an important factor. PMID- 19559273 TI - Is a 1-cm margin from major vessels adequate for radiofrequency ablation of pulmonary neoplasms? PMID- 19559275 TI - Paracorporeal artificial lung circuit as a possibility for bridge to lung transplantation. PMID- 19559277 TI - Vascular tumors of the sternum. PMID- 19559278 TI - MRI for preterm infants. PMID- 19559279 TI - Pediatricians on the front line for depression in children and parents. PMID- 19559280 TI - Hyperglycemia during induction of acute lymphocytic leukemia. PMID- 19559281 TI - Improving on quality adjusted life years measurements for health outcomes. PMID- 19559282 TI - Nephrosis and clots. PMID- 19559283 TI - Cardiovascular effects of methylphenidate. PMID- 19559284 TI - Morbidity and ambulatory resource utilization for diarrheal diseases. PMID- 19559285 TI - Residency research requirement as a predictor of future publication productivity. PMID- 19559286 TI - Reflections on the past, present, and future of the Journal of Pediatrics. PMID- 19559287 TI - Looking ahead to the next 75 years for the Journal of Pediatrics and medical publishing. PMID- 19559288 TI - Current challenges and future research in measuring preferences for pediatric health outcomes. PMID- 19559289 TI - Prenatal diagnosis of congenital heart disease in an era of near-universal ultrasound screening: room for improvement. PMID- 19559290 TI - Hypoplastic left heart syndrome: a paradigm case for examining conscientious objection in pediatric practice. PMID- 19559295 TI - Dent-2 disease: a mild variant of Lowe syndrome. AB - OBJECTIVE: To compare the renal and extra-renal phenotypes of patients classified as having Dent disease, Dent-2 disease, or Lowe syndrome. STUDY DESIGN: Chart review of data from 93 patients with identified voltage-gated chloride channel and chloride/proton antiporter 5 gene and oculo-cerebro-renal syndrome of Lowe gene mutations observed by the authors, complemented with published data. RESULTS: There was a wide overlap of renal symptoms. Nephrocalcinosis was more prevalent in Dent-1 disease, and renal tubular acidosis, aminoaciduria, and renal failure was more prevalent in patients with Lowe syndrome. Patients with Lowe syndrome were shorter than patients with Dent-1 disease, and patients with Dent-2 disease showed an intermediate phenotype. Three patients with Dent-2 disease had mild peripheral cataract, and 9 patients were noted to have some degree of mental retardation. CONCLUSION: There is a phenotypic continuum within patients with Dent-2 disease and Lowe syndrome, suggesting that there are individual differences in the ability to compensate for loss of oculo-cerebro-renal syndrome of Lowe gene function. PMID- 19559297 TI - A randomized controlled trial to evaluate the efficacy of lactobacillus GG in infantile diarrhea. AB - In a randomized, double-blind, placebo-controlled trial, 229 infants hospitalized for acute diarrhea in rural India were given a 10-day course of Lactobacillus rhammosus GG (minimum dose, 10 degrees bacteria) or placebo. There was no difference in groups in the duration of diarrhea or numbers of stool on days 3, 6, or 10 of treatment. PMID- 19559298 TI - The association between adiponectin/leptin ratio and diabetes type: the SEARCH for Diabetes in Youth Study. AB - We tested the association of adiponectin/leptin ratio with diabetes type after adjusting for multiple factors in 1156 youths with newly diagnosed diabetes in the SEARCH study. Although adiponectin/leptin ratio is associated with diabetes type in youth, it is due to differences in adiponectin, but not leptin levels. PMID- 19559301 TI - A novel MECP2 mutation in a boy with neonatal encephalopathy and facial dysmorphism. AB - Methly-CpG-binding protein 2 (MECP2) mutations cause Rett syndrome in females. Here we report on a male infant with neonatal encephalopathy, myoclonic jerks, and irregular breathing patterns caused by a novel frameshift mutation in the MECP2 gene. In addition he has facial dysmorphisms previously not described in these patients. PMID- 19559299 TI - Rituximab therapy for severe refractory chronic Henoch-Schonlein purpura. AB - To report on the efficacy of rituximab (RTX) therapy in standard treatment refractory, chronic Henoch-Schonlein purpura, a retrospective chart review of 3 pediatric patients treated with RTX for severe refractory chronic Henoch Schonlein purpura was performed. All 3 patients responded to 1 or 2 courses of RTX without serious adverse events. PMID- 19559302 TI - Recurrent unexplained episodes of facial cyanosis and shortness of breath in Hunter disease. PMID- 19559303 TI - Pediatric cardiologists can accurately use physical examination to identify pathologic murmurs in neonates. PMID- 19559304 TI - Elbow extension test is helpful in decreasing the likelihood of an elbow fracture in children. PMID- 19559305 TI - Universal primer polymerase chain reaction can diagnose neonatal sepsis when performed before starting antibiotics. PMID- 19559306 TI - E-mail intervention decreases online health risk references among adolescents. PMID- 19559307 TI - Pulse oximetry before discharge from the nursery can increase detection of serious congenital heart disease. PMID- 19559308 TI - Adjustment of cerebrospinal fluid cell counts for a traumatic lumbar puncture does not aid diagnosis of meningitis in neonates. PMID- 19559309 TI - Pneumococcal vaccine reduces the rates of pneumococcal meningitis in children. PMID- 19559310 TI - Efficacy of applied behavior analysis in autism. PMID- 19559311 TI - Takayasu's aortitis and infliximab. PMID- 19559313 TI - Liver enzymes in children and adolescents with eating disorders. PMID- 19559315 TI - Fetal surgery is a multidisciplinary field that relies on the participation and expertise of a variety of clinicians and support staff. Preface. PMID- 19559316 TI - Drugs of choice for sedation and analgesia in the neonatal ICU. AB - Painful procedures in the neonatal ICU are common, undertreated, and lead to adverse consequences. The drugs most commonly used to treat neonatal pain include the opiates, benzodiazepines, barbiturates, ketamine, propofol, acetaminophen, and local and topical anesthetics. This article discusses the indications for and advantages and disadvantages of the commonly used analgesic drugs. Guidance and references for drugs and dosing for specific neonatal procedures are provided. PMID- 19559317 TI - A history of fetal surgery. AB - Over the past 3 decades, fetal surgery for congenital disease has evolved from merely a fanciful concept to a medical field in its own right. Techniques for open hysterotomy, minimal-access hysteroscopy, and image-guided percutaneous fetal access have become well established, first in animal models and subsequently in humans. At the same time, major advances in fetal imaging and diagnosis, anesthesia, and tocolysis have allowed fetal intervention to become a vital tool for subsets of patients who would otherwise endure significant morbidity and mortality. This article offers a concise overview of the history of fetal surgery, from its tumultuous early days to its current status as an important means for the early treatment of potentially devastating congenital anomalies. PMID- 19559318 TI - Ethics of fetal surgery. AB - This article provides a comprehensive approach to the ethics of clinical investigation of fetal surgery. Investigators should address the initiation and assessment of clinical trials to determine whether they establish a standard of care and use an appropriate informed consent process to recruit and enroll subjects, consider whether selection criteria should include the abortion preferences of the pregnant woman, and consider whether physicians have an obligation to offer referral to such investigation. This approach is comprehensive because it takes account of the physician's obligations to the fetal patient, the pregnant woman, and future fetal and pregnant patients. The comprehensive approach to the ethics of fetal surgery is applied to the example of in utero surgical management of spina bifida. PMID- 19559319 TI - The maternal side of maternal-fetal surgery. AB - The term fetal surgery is used widely for fetal intervention during pregnancy; maternal-fetal surgery may be more appropriate, because all these invasive procedures also affect the mother. Although there is no direct benefit to the mother from these procedures, the risk to her is for a purely altruistic purpose. It is therefore important to understand the potential complications of maternal fetal surgery, so the physician can provide accurate counseling to the patient. PMID- 19559320 TI - The use of ultrasound in fetal surgery. AB - Obstetric ultrasound (US) is an integral part of fetal surgery for open and minimally invasive techniques. With advances in US imaging, the ability to refine diagnosis, predict prognosis, and contribute to fetal treatment continues to grow. Current research in fetal diagnosis and treatment includes identifying the most reliable sonographic features for determining prognosis before and after surgery. PMID- 19559321 TI - MRI of the fetal central nervous system and body. AB - MRI is being increasingly used to assess for fetal abnormalities. Although significant progress in the field of fetal MRI has occurred during the past 20 years, continued technical advances will likely contribute to significant growth of the field. Moreover, with continued hardware and software improvements, additional MRI sequences will likely become available. Prenatal MRI complements ultrasound because of larger field-of-view, superior soft tissue contrast, easier and more precise volumetric measurement, and greater accuracy in the demonstration of intracranial and spinal abnormalities. While ultrasound remains the primary modality for fetal imaging, these advantages of MRI make it a valuable adjunct to fetal surgery. Because fetal MRI involves many disciplines, the future of fetal MR will best be achieved through collaborative efforts. PMID- 19559322 TI - The role of fetal echocardiography in fetal intervention: a symbiotic relationship. AB - In this review, the authors explore the role of noninvasive and invasive fetal interventions in fetal cardiovascular disease guided by observations at fetal echocardiography. They first review fetal cardiac lesions that may be ameliorated by fetal intervention and then review noncardiac fetal pathologic findings for which fetal echocardiography can provide important insight into the pathophysiology and aid in patient selection for and timing of intervention and postintervention surveillance. PMID- 19559323 TI - Changing perspectives on the perinatal management of isolated congenital diaphragmatic hernia in Europe. AB - Congenital diaphragmatic hernia (CDH) should be diagnosed in the prenatal period and prompt referral to a tertiary referral center for imaging, genetic testing, and multidisciplinary counseling. Individual prediction of prognosis is based on the absence of additional anomalies, lung size, and liver herniation. In severe cases, a prenatal endotracheal balloon procedure is currently being offered at specialized centers. Fetal intervention is now also offered to milder cases within a trial, hypothesizing that this may reduce the occurrence of bronchopulmonary dysplasia in survivors. Postnatal management has been standardized by European high-volume centers for the purpose of this and other trials. PMID- 19559324 TI - Tracheal occlusion for fetal congenital diaphragmatic hernia: the US experience. AB - Congenital diaphragmatic hernia (CDH) is characterized by a defect in the diaphragm that permits abdominal viscera to herniate into the chest. These herniated viscera are thought to compress the growing lung and cause lung parenchymal and vascular hypoplasia. The genetic defects that cause the diaphragmatic defect may also contribute primarily to lung hypoplasia. Postnatal reduction of the herniated abdominal viscera and correction of the diaphragmatic defect are easily achievable, but the lung hypoplasia persists, often leading to persistent fetal circulation and respiratory failure. This article reviews the experimental basis of fetal therapy for CDH and the US clinical experience with tracheal occlusion. PMID- 19559325 TI - Management of fetal lung lesions. AB - Prenatal diagnosis provides insight into the in utero evolution of fetal thoracic lesions such as congenital cystic adenomatoid malformation (CCAM), bronchopulmonary sequestration (BPS), congenital lobar emphysema, and mediastinal teratoma. Serial sonographic study of fetuses with thoracic lesions has helped define the natural history of these lesions, determine the pathophysiologic features that affect clinical outcome, and formulate management based on prognosis. PMID- 19559326 TI - Fetal lower urinary tract obstruction. AB - The authors present an overview of the prenatal diagnosis, evaluation, contemporary intervention, and antenatal management of lower urinary tract obstruction. They review early experimental models that confirmed the relation between urinary tract obstruction and renal fibrocystic dysplasia and that early in utero relief of the obstruction could prevent irreversible renal injury. Subsequent studies of the electrolyte and protein concentrations in fetal urine from human cases established prognostic threshold values and helped to develop an algorithm to select candidates for antenatal therapy. Although shunting has improved survival, long-term morbidities remain a significant challenge. PMID- 19559327 TI - Twin-to-twin transfusion syndrome: a comprehensive update. AB - Twin-to-twin transfusion syndrome (TTTS) is a serious complication in about 10% to 20% of monozygous twin gestations with an incidence of 4% to 35% in the United States. Severe TTTS is reported to occur in 5.5% to 17.5% of cases. TTTS is a progressive disease in which sudden deteriorations in clinical status can occur, leading to death of a co-twin. Up to 30% of survivors may have abnormal neurodevelopment as a result of the combination of profound antenatal insult and the complications of severe prematurity. This article presents an overview of what is known about the pathophysiology and the diagnosis of TTTS, the role of echocardiography in TTTS, treatment options available for TTTS, complications of treatment for TTTS, and short- and long-term outcomes of TTTS. PMID- 19559328 TI - Complicated monochorionic twin pregnancies: updates in fetal diagnosis and treatment. AB - Monochorionic (MC) twin pregnancies may develop significant complications, and twin-to-twin transfusion syndrome (TTTS) has become among the best known to obstetricians and patients alike. A significant percentage of patients referred for suspected TTTS have a different underlying pathologic condition, however, and differentiating the subcategories of MC pathophysiologic conditions may change treatment course and outcome. The key to understanding complicated MC pregnancies lies in the placental angioarchitecture and intertwin vascular communications between the fetuses. PMID- 19559329 TI - Fetal surgery for myelomeningocele. AB - Fetal intervention for myelomeningocele (MMC) may improve hydrocephalus and hindbrain herniation associated with the Arnold-Chiari II malformation and may reduce the need for ventriculoperitoneal shunting. As of now, there is little evidence that prenatal repair of MMC improves neurologic function. MMC is the first nonlethal disease under consideration and study for fetal surgery. As a result, potential improvements in outcome must be balanced with maternal safety and well-being, in addition to that of the unborn patient. PMID- 19559330 TI - Cardiac anomalies in the fetus. AB - Congenital heart disease (CHD) is an attractive target for fetal therapy. With the development of successful neonatal repair for many types of CHD over the last 20 years, fetal therapy has become the next frontier. Concurrent advances in interventional catheterization and fetal imaging provided a foundation for the novel field of fetal cardiac intervention. This article focuses on the current status of in utero catheter interventions for CHD with particular interest in therapy for defects characterized by progressive stenosis or atresia of the semilunar valves, the aortic and pulmonary, with development of subsequent ventricular hypoplasia. PMID- 19559331 TI - Prenatal stem cell transplantation and gene therapy. AB - At the present time, the most likely and eminent application of stem cell therapy to the fetus is in utero hematopoietic stem cell transplantation (IUHCT), and this stem cell type will be discussed as a paradigm for all prenatal stem cell therapy. The authors feel that the most likely initial application of IUHCT will use adult HSC derived from bone marrow (BM) or peripheral blood (PB), and will focus this article on this specific approach. The article also reviews the experimental data that support the capacity of IUHCT to induce donor-specific tolerance. PMID- 19559332 TI - Fetal tissue engineering. AB - Attempts at harnessing the prospective benefits of the therapeutic use of fetal cells or tissues date many decades before the modern era of transplantation. The first reported transplantation of human fetal tissue took place in 1922. Fetal cells or tissues also have been used as helpful investigational tools since the 1930s. Still, it was only in the last three decades that fetal tissue transplantation in people has started to lead to favorable outcomes, yet by and large anecdotally. This article offers an outlook on a relatively new dimension in fetal cell-based therapies, namely the engineering of tissues in the laboratory, along with its prospective applications. PMID- 19559333 TI - Quiz page July 2009. Fever and neutropenia following simultaneous pancreas-kidney transplantation. Graft-versus-host disease. PMID- 19559334 TI - CKD prognosis: beyond the traditional outcomes. PMID- 19559335 TI - Improving training in nephrology procedures: yes we can. PMID- 19559336 TI - Overcoming challenges to influenza vaccination in patients with CKD. PMID- 19559337 TI - Treatment guidelines for dialysis catheter-related bacteremia: an update. PMID- 19559338 TI - Thirty-six Hippocratic aphorisms of nephrologic interest. AB - In this article, we discuss the nephrologic content within Hippocrates' Aphorisms. Although similar attempts have taken place ever since antiquity, we believe that in each era new insights may be gained by examining the aphorisms through the prism of current medical knowledge. Of the 400 aphorisms in the Hippocratic text, we discuss the 36 that we consider to be most relevant to nephrology. We conclude that these aphorisms support the concept of Hippocrates as the "Father of Clinical Nephrology." PMID- 19559339 TI - Restriction of dietary protein and long-term outcomes in patients with CKD. PMID- 19559340 TI - Effect of a very low-protein diet on long-term outcomes. PMID- 19559341 TI - Prevalence of CKD in the United States: GFR-estimating equations matter. PMID- 19559343 TI - Beyond amygdala: Default Mode Network activity differs between patients with social phobia and healthy controls. AB - The Default Mode Network (DMN) is a constellation of brain areas that decrease their activity during a wide number of different goal-oriented tasks as compared to passive "rest" tasks. DMN can be modulated by different factors such as emotional states, cognitive load of the task and psychopathology, including anxiety. Moreover, DMN seems to play a pivotal role in social cognition. For example, the ability to predict another person's behaviour taking his or her perspective modulates the activity of the DMN. Recent data from autistic patients support a role of DMN in social cognition as well. Social Phobia (SP) is an anxiety disorder characterized by an abnormal distress in situations that require social interaction. To date, no study has assessed DMN in Social Phobia. To determine potential differences in DMN activity between Social Phobia patients (SPP) and healthy control (HC) subjects we examined functional magnetic resonance imaging (fMRI) data obtained during a face perception study with emotional and neutral stimuli. As compared to HC, SPP showed a lower deactivation in the precuneus and posterior cingulate regions (PCun/PCC) during task conditions. These regions are part of the so-called "Theory of Mind" circuit and in particular they are involved in the evaluation of one's own emotional state. Because of the role of the PCun/PCC in self-state perception and attribution and, more in general, the role of the DMN in social cognition, we suggest that its impairment in the DMN network in SPP might be relevant in the development of the feeling of wariness of others' judgment and may be related to the so-called self focused attention. Self-focused attention is the awareness of self-referent information, and is present in many emotional disorders and may additionally prevent individuals from observing external information that could disconfirm their own fears. Moreover, the abnormal modulation of activity in the DMN may reflect persistent rumination or anxiety-related thoughts that are not modulated by the switch from rest to task. PMID- 19559344 TI - Genetic variants on chromosome 9p21 and ischemic stroke in Chinese. AB - In a hospital based case control study, we investigated the association of cyclin dependent kinase inhibitor 2A (CDKN2A) gene, cyclin-dependent kinase inhibitor 2B (CDKN2B) gene, and two genetic variants (rs10757274 and rs2383206) on chromosome region 9p21 with ischemic stroke in Chinese Hans. Two polymorphisms in the CDKN2A gene (rs3088440 and rs3731245) and two polymorphisms in the CDKN2B gene (rs3217992 and rs1063192) were selected by using a strategy of tagging single nucleotide polymorphisms (tSNP). We observed significant association of rs2383206 with ischemic stroke. Subjects with the GG/GA genotype of rs2383206 had a 1.51 fold (95%CI 1.11-2.05, p=0.009) increased risk of stroke, compared with those with the AA genotype. In addition, the GG/GA genotypes of rs2383206 and rs3731245 was associated with an increased risk of large vessel subtype and small vessel subtype of ischemic stroke, respectively, with ORs of 2.09 (95%CI 1.30-3.37, p=0.002) and 1.63 (95%CI 1.06-2.51, p=0.026), respectively. In gene-environmental interaction analysis, elevation of ischemic stroke risk was observed among AG+GG genotype carriers who consume alcohol, smoke cigarette, and have hypertension, with adjusted combined ORs of 2.86(1.51-5.41), 4.30(2.38-7.77), and 13.97(7.78 25.07), respectively, compared with low-risk individuals for rs2383206 (GG carriers who did not consume alcohol, smoke cigarette, and without hypertension). We provide evidence that genetic variants on chromosome region 9p21 may implicated in the prevalence of ischemic stroke in Chinese. PMID- 19559345 TI - Evidence for 2-stage models of sleep and memory: learning-dependent changes in spindles and theta in rats. AB - What processes are involved in the formation of enduring memory traces? Sleep has been proposed to play a role in memory consolidation and the present study provides evidence to support 2-stage models of sleep and memory including both non-rapid eye movement (NREM) and rapid eye movement (REM) sleep. Previous research has shown REM sleep increases following avoidance learning and memory is impaired if REM deprivation occurs during these post-training periods indicating that REM sleep may have a role in memory consolidation processes. These discrete post-training periods have been termed REM sleep windows (RSWs). It is not known whether the electroencephalogram has unique characteristics during the RSW. Further investigation of the RSW was one of the primary goals of this study. We investigated the epidural-recorded electrophysiological learning-related changes following avoidance training in rats. Theta power increased in the learning group during the RSW, suggesting that theta is involved in memory consolidation during this period. Sleep spindles subsequently increased in slow wave sleep (SWS). The results suggest that both NREM and REM sleep are involved in sleep-dependent memory consolidation, and provide support for existing 2-stage models. Perhaps first theta increases to organize and consolidate material via hippocampal neocortical dialogue, followed by subsequent refinement in the cortex by spindles during SWS. PMID- 19559346 TI - Abdominal complications of chemotherapy in pediatric malignancies: imaging findings. AB - OBJECTIVE: To describe the imaging findings of abdominal complications caused by chemotherapy in pediatric cancer patients. SUBJECTS AND METHODS: Radiology studies of 243 patients treated in our Children Cancer Center were reviewed, 164 of whom had abdominal studies. Medical records of 54 patients with abnormal imaging findings were studied. RESULTS: Ultrasound and/or CT findings showed the following complications: (1) gallbladder sludge/crystals (n=13), gallstones (n=8), cholecystitis (n=3); (2) liver steatosis (n=23), siderosis (n=1), veno occlusive disease (n=2); (3) pancreatitis (n=7); (4) typhlitis (n=12), esophagitis (n=2). CONCLUSION: Awareness of these complications is essential for appropriate management, decreasing their mortality and morbidity. PMID- 19559347 TI - Use of a saline chaser in abdominal computed tomography: a systematic review. AB - PURPOSE: To perform a systematic review of the literature regarding the use of a saline chaser in abdominal computed tomography (CT). MATERIALS AND METHODS: The MEDLINE database was searched from 1966 to March 2007. Studies were included if they compared the magnitude of contrast enhancement with and without a saline chaser in CT and reported CT contrast enhancement values of the liver, portal vein, or abdominal aorta. For randomized controlled trials (RCTs) comparing the same dose of contrast material with and without a saline chaser, mean differences in the magnitude of contrast enhancement were pooled by using a fixed-effects model. RESULTS: Seventeen studies met the inclusion criteria. Three RCTs using clinical images were included in the meta-analysis. The mean differences in contrast enhancement of the liver, portal vein, and aorta were -0.4 (95% CI, -2.4 to 1.7), 7.4 (95% CI, 1.3 to 14), and 11 (95% CI, -5.3 to 27), respectively. Three RCTs using a time-density analysis were included in the meta-analysis. The mean differences in peak contrast enhancement of the liver, portal vein, and aorta were 8.2 (95% CI, 4.2 to 12), 28 (95% CI, 16 to 40), and 15 (95% CI, 0.3 to 30), respectively. CONCLUSION: In clinical images, a saline chaser did not improve contrast enhancement of the liver. In time-density analysis, however, a saline chaser improved peak contrast enhancement of the liver. PMID- 19559349 TI - CT findings of mild forms or early manifestations of acute cholecystitis. AB - OBJECTIVE: The objective of this study was to determine the most predictive CT feature of the mild forms or early manifestations of acute cholecystitis. MATERIALS AND METHODS: Two radiologists analyzed CT of 34 patients with mild or early acute cholecystitis and 34 control patients for pericholecystic increased attenuation on the arterial phase, indistinctness of the interface between the gallbladder (GB) and the liver, enhancement of the GB wall, and increased attenuation of the GB bile. RESULTS: There were significant differences in the mean values for each CT feature but increased attenuation of the GB bile between patients and control group (P<.05). The most significant predictor of mild or early acute cholecystitis on CT was the presence of pericholecystic increased attenuation on the arterial phase (sensitivity, 82.4%), followed by indistinctness of the interface between the GB and liver (sensitivity, 38.0%), which were identified by both observers with good agreement (kappa=0.735 and kappa=0.687). CONCLUSIONS: The pericholecystic increased attenuation on arterial phase CT was the most significant predictor of mild forms or early manifestations of acute cholecystitis. PMID- 19559350 TI - Detection and characterization of focal hepatic lesions by T2-weighted imaging: comparison of navigator-triggered turbo spin-echo, breath-hold turbo spin-echo, and HASTE sequences. AB - The aim of this study is to evaluate the diagnostic performance of T(2)-weighted (T(2)-w) images obtained using navigator-triggered turbo spin-echo (TSE), breath hold TSE (BH-TSE), and BH haft-Fourier single-shot TSE sequences for the detection and characterization of focal hepatic lesions. Two blinded reviewers independently analyzed three types of T(2)-w image sets totaling 86 solid and 75 nonsolid lesions in 59 patients. Receiver operating characteristic curves were established to analyze reviewer and sequence results. PMID- 19559348 TI - Magnetic resonance imaging of the small bowel with the true FISP sequence: intra- and interobserver agreement of enteroclysis and imaging without contrast material. AB - PURPOSE: This study aimed to determine the reliability of magnetic resonance imaging (MRI) without luminal contrast medium versus MR enteroclysis for evaluating small bowel pathology, to compare MRI and MRE findings per observer, and to compare these findings with those of an expert reader in order to determine the influence of luminal contrast medium on morphological evaluations. CONCLUSION: The use of luminal contrast medium bowel improves reliability for measuring bowel wall thickness and for the diagnosis and grading of obstruction when evaluating the small bowel. PMID- 19559351 TI - Liver perfusion with dynamic multidetector-row computed tomography as an objective method to evaluate the efficacy of chemotherapy in patients with colorectal cancer. AB - A total of 24 liver metastases of colorectal cancer were evaluated by dynamic multiphasic CT. Under chemotherapy, follow-up examinations were performed every 3 months. The tumor marker CEA before vs. after chemotherapy correlated with the mean contrast enhancement within the margin of metastases. The total size of metastases correlated with the size of central necrosis as well as with the size of marginal contrast enhancement. CT is able to quantify perfusion and local activity of liver metastases to determine the efficacy of chemotherapy. PMID- 19559353 TI - Lymphotropic nanoparticle-enhanced magnetic resonance imaging (LNMRI) identifies occult lymph node metastases in prostate cancer patients prior to salvage radiation therapy. AB - Twenty-six patients with prostate cancer status post-radical prostatectomy who were candidates for salvage radiation therapy (SRT) underwent lymphotropic nanoparticle enhanced MRI (LNMRI) using superparamagnetic nanoparticle ferumoxtran-10. LNMRI was well tolerated, with only two adverse events, both Grade 2. Six (23%) of the 26 patients, previously believed to be node negative, tested lymph node positive by LNMRI. A total of nine positive lymph nodes were identified in these six patients, none of which were enlarged based on size criteria. PMID- 19559352 TI - The characterization of small hypoattenuating renal masses on contrast-enhanced CT. AB - PURPOSE: To determine if small hypoattenuating renal masses can be characterized as simple cysts or renal cell carcinomas on contrast-enhanced computed tomography (CT). MATERIALS AND METHODS: We retrospectively identified 20 small (or=6, and the improvement index was greater or equal to +1 in 67% of case. CONCLUSION: In this initial monocentric experiment, despite a modest improvement of objective parameters and a 20% of retreatment rate, TUNA give contentment and improvement sensation for 60% of patients who were treated for non-efficiency of medical treatment for benign prostatic hyperplasia. PMID- 19559384 TI - [Urethroplasty for hypospadias: assessing the urethral patch]. PMID- 19559385 TI - [Retrograde endopyelotomy for stenosis of the renal pelvis and ureter junction]. PMID- 19559386 TI - [Domino transplantation of a living-donor kidney graft affected by thrombotic microangiopathy: surgical aspects]. AB - We report surgical aspects of the first case of retransplantation of a kidney initially retrieved from a cadaveric donor, then on a first recipient which developed a recurrent severe and intractable thrombotic microangiopathy on the allograft. PMID- 19559387 TI - Minor painful procedures in the NICU: improved care or too soon for cheering? PMID- 19559388 TI - Genetic contributions to clinical pain and analgesia: avoiding pitfalls in genetic research. AB - Understanding the genetic basis of human variations in pain is critical to elucidating the molecular basis of pain sensitivity, variable responses to analgesic drugs, and, ultimately, to individualized treatment of pain and improved public health. With the help of recently accumulated knowledge and advanced technologies, pain researchers hope to gain insight into genetic mechanisms of pain and eventually apply this knowledge to pain treatment. PERSPECTIVE: We critically reviewed the published literature to examine the strength of evidence supporting genetic influences on clinical and human experimental pain. Based on this evidence and the experience of false associations that have occurred in other related disciplines, we provide recommendations for avoiding pitfalls in pain genetic research. PMID- 19559389 TI - Local antinociception induced by endothelin-1 in the hairy skin of the rat's back. AB - Subcutaneous injection of endothelin-1 (ET-1) into the glabrous skin of the rat's hind paw is known to produce impulses in nociceptors and acute nocifensive behavioral responses, such as hind paw flinching, and to sensitize the skin to mechanical and thermal stimulation. In this report, we show that in contrast to the responses in glabrous skin, ET-1 injected subcutaneously into rat hairy skin causes transient antinociception. Concentrations of 1 to 50 microM ET-1 (in 0.05 mL) depress the local nocifensive response to noxious tactile probing at the injection site with von Frey filaments for 30 to 180 minutes; distant injections have no effect at this site, showing that the response is local. Selective inhibition of ET(A) but not of ET(B) receptors inhibits this antinociception, as does coinjection with nimodipine (40 muM), a blocker of L-type Ca(2+) channels. Local subcutaneous injection of epinephrine (45 microM) also causes antinociception through alpha-1 adrenoreceptors, but such receptors are not involved in the ET-1-induced effect. Both epinephrine and ET-1, at antinociceptive concentrations, reduce blood flow in the skin; the effect from ET 1 is largely prevented by subcutaneous nimodipine. These data suggest that ET-1 induced antinociception in the hairy skin of the rat involves cutaneous vasoconstriction, presumably through neural ischemia, resulting in conduction block. PERSPECTIVE: The pain-inducing effects of ET-1 have been well documented in glabrous skin of the rat, a frequently used test site. The opposite behavioral effect, antinociception, occurs from ET-1 in hairy skin and is correlated with a reduction in blood flow. Vasoactive effects are important in assessing mechanisms of peripherally acting agents. PMID- 19559390 TI - Early and late contributions of glutamate and CGRP to mechanical sensitization by endothelin-1. AB - Intraplantar injection of endothelin-1 (ET-1) (1.5-10 muM) in the rat produces mechanical allodynia. Here we identify the receptor subtypes for ET-1, glutamate and CGRP critical to such allodynia. Antagonism of ET(A) or ET(B) receptors alone, by BQ123 or BQ788, respectively, only partially suppressed allodynia; the combined antagonists prevented allodynia, showing the involvement of both receptor subtypes. Co-injection of NMDA receptor antagonists, (+)MK-801 or D-AP5, with ET-1 also prevented allodynia. In contrast, co-injection of the CGRP1 antagonist CGRP(8-37) attenuated only the later phase of allodynia (>30 min). A mechanistic basis for these effects is shown by ET-1's ability to enhance basal release from cultured sensory neurons of glutamate and CGRP (2.4-fold and 5.7 fold, respectively, for 10 nM ET-1). ET(A) blockade reduced ET-1's enhancement of basal CGRP release by approximately 80%, but basal glutamate release by only approximately 30%. ET-1 also enhanced the capsaicin-stimulated release of CGRP (up to 2-fold for 0.3 nM ET-1), but did not change capsaicin-stimulated glutamate release. Release stimulated by elevated K+ was not altered by ET(A) blockade, nor did blockade of ET(B) reduce any type of release. Thus, ET-1 may induce release of glutamate and CGRP from nerve terminals innervating skin, thereby sensitizing primary afferents, accounting for ET-1-dependent tactile allodynia. PERSPECTIVE: The endogenous endothelin peptides participate in a remarkable variety of pain related processes. The present results provide evidence for the participation of ionotropic glutamatergic receptors and CGRP receptors in the hyperalgesic responses to exogenous ET-1 and suggest clinically relevant targets for further study of elevated pain caused by release of endogenous ET-1. PMID- 19559391 TI - Sodium channel expression and localization at demyelinated sites in painful human dental pulp. AB - The expression of sodium channels (NaCh(s)) change after inflammatory and nerve lesions, and this change has been implicated in the generation of pain states. Here we examine NaCh expression within nerve fibers from normal and painful extracted human teeth with special emphasis on their localization within large accumulations, like those seen at nodes of Ranvier. Pulpal tissue sections from normal wisdom teeth and from teeth with large carious lesions associated with severe and spontaneous pain were double-stained with pan-specific NaCh antibody and caspr (paranodal protein used to visualize nodes of Ranvier) antibody, while additional sections were triple-stained with NaCh, caspr and myelin basic protein (MBP) antibodies. Z-series of images were obtained with the confocal microscope and evaluated with NIH ImageJ software to quantify the density and size of NaCh accumulations, and to characterize NaCh localization at caspr-identified typical and atypical nodal sites. Although the results showed variability in the overall density and size of NaCh accumulations in painful samples, a common finding included the remodeling of NaChs at atypical nodal sites. This remodeling of NaChs included prominent NaCh expression within nerve regions that showed a selective loss of MBP staining in a pattern consistent with a demyelinating process. PERSPECTIVE: This study identifies the remodeling of NaChs at demyelinated sites within the painful human dental pulp and suggests that the contribution of NaChs to spontaneous pulpal pain generation may be dependant not only on total NaCh density but may also be related to NaCh expression at atypical nodal sites. PMID- 19559393 TI - Methadone for pain: limited evidence, tenuous guidelines. PMID- 19559395 TI - Abstracts of ICCBH5, Fifth International Conference on Children's Bone Health. Cambridge, United Kingdom. June 23-26, 2009. PMID- 19559392 TI - A candidate gene association study of 77 polymorphisms in migraine. AB - Population-based studies have established an association between migraine and cardiovascular disease (CVD). We sought to investigate whether genetic variants implicated in CVD are associated with migraine. We performed an association study among 25,713 women participating in the Women's Health Study, with information on 77 previously characterized polymorphisms. Migraine and migraine aura status were self-reported. We used logistic regression to investigate the genotype-migraine association. At baseline, 4705 (18.3%) women reported history of migraine; 39.6% of the 3306 women with active migraine indicated aura. Regarding any history of migraine, the multivariable-adjusted odds ratios (95% confidence intervals) for TNF rs673 were 0.52 (0.30 to 0.89), for TGFB1 rs1800469 0.93 (0.89 to 0.98), and for CCR2 rs1799864 1.12 (1.03 to 1.21). Among active migraine with aura, the odds ratios (95% confidence intervals) were 1.35 (1.0 to 1.81) for TNF rs1800750, 1.13 (1.02 to 1.26) for TNF rs1800629, and 1.22 (1.07 to 1.40) for CCR2 rs1799864; among active migraine without aura, 0.9 (0.84 to 0.97) for TGFB1 rs1800469, 1.13 (1.01 to 1.27) for NOS3 rs3918226, and 1.12 (1.02 to 1.24) for IL9 rs2069885. After correction for multiple testing using the false discovery rate, none of the results remained significant. Our data suggest an association of polymorphisms implicated in inflammatory pathways and migraine in women. TNF, CCR2, TGFB1, NOS3, and IL9 warrant further investigation. PERSPECTIVE: This article presents results from an association study of 77 polymorphisms, implicated in CVD, and migraine. Variants in TNF, CCR2, TGFB1, NOS3, and IL9 were found to be associated with migraine but did not remain significant after adjustment for multiple testing. Variations in these genes warrant further investigation. PMID- 19559396 TI - Automatic sequential response priming and intentional response preparation in choice reaction tasks: evidence from response repetition and response cuing. AB - This study examined the interaction of response repetition and response cuing in a finger cuing task with a short and a long cue-stimulus interval (CSI). We observed shorter reaction times (RTs) with increasing CSI and a substantial response repetition benefit. However, this benefit was abolished at the long CSI, suggesting that response cuing neutralized the repetition effect. According to additive-factors logic, the observed interaction suggests that both repetition and cuing exert their influence on a common processing stage, which we identify as the response selection stage. We argue that cuing and repetition effects are expressions of distinct mental operations: cuing is based on intentional response code activation, whereas repetition is based on sequential, automatic response code priming. Cue-based intentional code activation starts slowly and increases with CSI, but sequential response priming is independent of CSI, explaining why cuing abolishes the response repetition benefit at the long CSI. PMID- 19559397 TI - Mutation in the AP4M1 gene provides a model for neuroaxonal injury in cerebral palsy. AB - Cerebral palsy due to perinatal injury to cerebral white matter is usually not caused by genetic mutations, but by ischemia and/or inflammation. Here, we describe an autosomal-recessive type of tetraplegic cerebral palsy with mental retardation, reduction of cerebral white matter, and atrophy of the cerebellum in an inbred sibship. The phenotype was recorded and evolution followed for over 20 years. Brain lesions were studied by diffusion tensor MR tractography. Homozygosity mapping with SNPs was performed for identification of the chromosomal locus for the disease. In the 14 Mb candidate region on chromosome 7q22, RNA expression profiling was used for selecting among the 203 genes in the area. In postmortem brain tissue available from one patient, histology and immunohistochemistry were performed. Disease course and imaging were mostly reminiscent of hypoxic-ischemic tetraplegic cerebral palsy, with neuroaxonal degeneration and white matter loss. In all five patients, a donor splice site pathogenic mutation in intron 14 of the AP4M1 gene (c.1137+1G-->T), was identified. AP4M1, encoding for the mu subunit of the adaptor protein complex-4, is involved in intracellular trafficking of glutamate receptors. Aberrant GluRdelta2 glutamate receptor localization and dendritic spine morphology were observed in the postmortem brain specimen. This disease entity, which we refer to as congenital spastic tetraplegia (CST), is therefore a genetic model for congenital cerebral palsy with evidence for neuroaxonal damage and glutamate receptor abnormality, mimicking perinatally acquired hypoxic-ischemic white matter injury. PMID- 19559398 TI - WNT10A mutations are a frequent cause of a broad spectrum of ectodermal dysplasias with sex-biased manifestation pattern in heterozygotes. AB - Odonto-onycho-dermal dysplasia (OODD), a rare autosomal-recessive inherited form of ectodermal dysplasia including severe oligodontia, nail dystrophy, palmoplantar hyperkeratosis, and hyperhidrosis, was recently shown to be caused by a homozygous nonsense WNT10A mutation in three consanguineous Lebanese families. Here, we report on 12 patients, from 11 unrelated families, with ectodermal dysplasia caused by five previously undescribed WNT10A mutations. In this study, we show that (1) WNT10A mutations cause not only OODD but also other forms of ectodermal dysplasia, reaching from apparently monosymptomatic severe oligodontia to Schopf-Schulz-Passarge syndrome, which is so far considered a unique entity by the findings of numerous cysts along eyelid margins and the increased risk of benign and malignant skin tumors; (2) WNT10A mutations are a frequent cause of ectodermal dysplasia and were found in about 9% of an unselected patient cohort; (3) about half of the heterozygotes (53.8%) show a phenotype manifestation, including mainly tooth and nail anomalies, which was not reported before in OODD; and (4) heterozygotes show a sex-biased manifestation pattern, with a significantly higher proportion of tooth anomalies in males than in females, which may implicate gender-specific differences of WNT10A expression. PMID- 19559399 TI - Loss-of-function mutation in the dioxygenase-encoding FTO gene causes severe growth retardation and multiple malformations. AB - FTO is a nuclear protein belonging to the AlkB-related non-haem iron- and 2 oxoglutarate-dependent dioxygenase family. Although polymorphisms within the first intron of the FTO gene have been associated with obesity, the physiological role of FTO remains unknown. Here we show that a R316Q mutation, inactivating FTO enzymatic activity, is responsible for an autosomal-recessive lethal syndrome. Cultured skin fibroblasts from affected subjects showed impaired proliferation and accelerated senescence. These findings indicate that FTO is essential for normal development of the central nervous and cardiovascular systems in human and establish that a mutation in a human member of the AlkB-related dioxygenase family results in a severe polymalformation syndrome. PMID- 19559400 TI - Closing the gap: inverting the genetics curriculum to ensure an informed public. AB - Over the past 20 years, the focus of national efforts to improve K-12 science education has ranged from curriculum and professional development of teachers to the adoption of science standards and high-stakes testing. In spite of this work, students in the United States continue to lag behind their peers in other countries. This underperformance is true for genetics, as well as for science and math in general, and is particularly worrisome given the accelerating need for scientists and engineers in our increasingly technology-driven economy. A scientifically literate public is essential if citizens are to engage effectively with policymakers on issues of scientific importance. Perhaps nowhere is this conjunction more personally meaningful than in human genetics and medicine. Rapid changes in our field have the potential to revolutionize healthcare, but the public is ill prepared to participate in this transformation. One potential solution is to modernize the genetics curriculum so that it matches the science of the 21(st) century. This paper highlights changes in human genetics that support a curricular reorganization, outlines the problems with current genetics instruction, and proposes a new genetics curriculum. PMID- 19559401 TI - The consequences of effortful emotion regulation when processing distressing material: a comparison of suppression and acceptance. AB - The present study investigated the consequences of different forms of emotion regulation. Eighty nine healthy participants viewed a distressing video of the aftermath of road traffic accidents under either suppression (of both felt and expressed affect), acceptance, or no-regulation control instructions and the immediate and longer-term consequences on emotion, mood, and memory were examined. Suppression (relative to control) led to reduced subjective experience of fear when viewing the video, but did not alter electrodermal (EDA) or heart rate (HR) response. Subsequently, suppression led to a less marked subjective emotional reaction to positive but not negative emotional images, reduced free recall memory of the video, and a greater likelihood of experiencing zero intrusions of the video's content. Acceptance (relative to control) had no impact when viewing the video, was associated with a less marked increase in EDA activity in the 5 min period immediately after viewing the video, a more marked HR deceleration and EDA response to both positive and negative images, and elevated negative affect at one week follow-up. These findings suggest, contrary to the current clinical zeitgeist, that emotion suppression can successfully lead to an ongoing down-regulation of emotion and memory, whereas acceptance may elevate subsequent emotionality. PMID- 19559402 TI - Glutamate dysfunction in people with prodromal symptoms of psychosis: relationship to gray matter volume. AB - BACKGROUND: The glutamate model of schizophrenia proposes that altered glutamatergic neurotransmission is fundamental to the development of the disorder. In addition, its potential to mediate neurotoxicity raises the possibility that glutamate dysfunction could underlie neuroanatomic changes in schizophrenia. Here we determine whether changes in brain glutamate are present in subjects at ultra high risk of developing psychosis and whether these changes are related to reductions in cortical gray matter volume. METHODS: Twenty-seven individuals with an at-risk mental state and a group of 27 healthy volunteers underwent proton magnetic resonance spectroscopy and volumetric proton magnetic resonance imaging using a 3-Tesla scanner. Glutamate and glutamine levels were measured in anterior cingulate, left hippocampus, and left thalamus. These measures were then related to cortical gray matter volume. RESULTS: At-risk mental state (ARMS) subjects had significantly lower levels of glutamate than control subjects in the thalamus (p < .05) but higher glutamine in the anterior cingulate (p < .05). Within the ARMS group, the level of thalamic glutamate was directly correlated with gray matter volume in the medial temporal cortex and insula (p < .01). CONCLUSIONS: This study provides the first evidence that brain glutamate function is perturbed in people with prodromal signs of schizophrenia and that glutamatergic dysfunction is associated with a reduction in gray matter volume in brain regions thought to be critical to the pathogenesis of the disorder. These findings support the hypothesis that drugs affecting the glutamate system may be of benefit in the early stages of psychotic illness. PMID- 19559404 TI - Involvement of NOX2 in the development of behavioral and pathologic alterations in isolated rats. AB - BACKGROUND: Social stress leads to oxidative stress in the central nervous system, contributing to the development of mental disorders. Loss of parvalbumin in interneurons is an important feature of these diseases. We studied the role of the superoxide-producing nicotinamide adenosine dinucleotide phosphate (NADPH) oxidase 2 (NOX2) in rats exposed to social isolation. METHODS: Male rats were kept for 7 weeks in group or in social isolation (n = 6-10 per group). Behavioral tests, immunohistochemistry, and analysis of NOX2 expression were performed at the end of social isolation. Apocynin was given in the drinking water (5 mg/kg/day). RESULTS: NOX2 was below detection level in the brains of control animals, whereas it was highly expressed in isolated rats, particularly in nucleus accumbens and prefrontal cortex. Indirect markers of oxidative stress (oxidized nucleic acid 8-hydroxy-2'-deoxyguanosine, redox-sensitive transcription factor c-fos, and hypoxia-inducible factor-1alpha) were increased after social isolation in brain areas with high NOX2 expression. An increase in immunoreactive microglia suggested that oxidative stress could be in part due to NOX2 activation in microglia. In response to social isolation, rats showed increased locomotor activity, decreased discrimination, signs of oxidative stress in neurons, and loss of parvalbumin-immunoreactivity. Treatment of isolated rats with the antioxidant/NOX inhibitor apocynin prevented the behavioral and histopathological alterations induced by social isolation. CONCLUSIONS: Our data suggest that NOX2 derived oxidative stress is involved in loss of parvalbumin immunoreactivity and development of behavioral alterations after social isolation. These results provide a molecular mechanism for the coupling between social stress and brain oxidative stress, as well as potential new therapeutic avenues. PMID- 19559403 TI - Cingulate white matter neurons in schizophrenia and bipolar disorder. AB - BACKGROUND: Increased neuronal density in prefrontal, parietal, and temporal white matter of schizophrenia subjects is thought to reflect disordered neurodevelopment; however, it is not known if this cellular alteration affects the cingulate cortex and whether similar changes exist in bipolar disorder. METHOD: Eighty-two postmortem specimens (bipolar 15, schizophrenia 22, control 45) were included in this clinical study. Densities for two neuronal markers, neuron-specific nuclear protein (NeuN) and neuregulin 1 alpha (NRG), were determined in white matter up to 2.5 mm beneath the anterior cingulate cortex; density of NeuN immunopositive neurons (NeuN+) was also determined for a subset of cases in prefrontal cortex. Changes during normal development were monitored in a separate cohort of 14 brains. RESULTS: Both the schizophrenia and bipolar cohorts demonstrated a twofold increase in NeuN+ density in cingulate white matter; this effect could be attributed to approximately 25% of cases that exceeded the second standard deviation from control subjects. Similar changes were observed in prefrontal cortex. In contrast density of NRG expressing neurons was unaltered. Cases with increased NeuN+ densities in two-dimensional (2-D) counts also showed a pronounced, > fivefold elevation in NeuN+ nuclei per cubic millimeter. Additionally, the developmental cohort demonstrated a 75% decline in NeuN+ neuronal density during the first postnatal year but was stable thereafter. CONCLUSIONS: Increased neuronal density in white matter of cingulate cortex in schizophrenia provides further evidence that this alteration occurs in multiple cortical areas. Similar changes in some cases with bipolar illness suggest that the two disorders may share a common underlying defect in late prenatal or early postnatal neurodevelopment. PMID- 19559405 TI - Antioxidant properties of some different molecular weight chitosans. AB - Chitosan, a cationic polysaccharide, is widely employed as dietary supplement and in pharmacological and biomedical applications. Although numerous studies have focused on its applications as pharmaceutical excipients or bioactive reagents, relationships between molecular weight (Mr) and biological properties remain unclear. The focus of this study was on the antioxidant properties of several Mr chitosans. We measured the ability of seven Mr chitosans (CT1; 2.8 kDa, CT2; 17.0 kDa, CT3; 33.5 kDa, CT4; 62.6 kDa, CT5; 87.7 kDa, CT6; 604 kDa, CT7; 931 kDa) to protect plasma protein from oxidation by peroxyl radicals derived from 2,2' azobis (2-amidinopropane) dihydrochloride (AAPH). A comparison of the antioxidant action of high Mr chitosans (CT6-CT7) with that of low Mr chitosans (CT1-CT5) showed that low Mr chitosans (CT1-CT5) were more effective in preventing the formation of carbonyl groups in plasma protein exposed to peroxyl radicals. AAPH substantially increases plasma protein carbonyl content via the oxidation of human serum albumin (HSA). We also measured the ability of these chitosans to protect HSA against oxidation by AAPH. Low Mr chitosans (CT1-CT5) were found to effectively prevent the formation of carbonyl groups in HSA, when exposed to peroxyl radicals. Low Mr chitosans were also good scavengers of N-centered radicals, but high Mr chitosans were much less effective. We also found a strong correlation between antioxidant activity and the Mr of chitosans in vitro. These activities were also determined by using the 'TPAC' test. These results suggest that low Mr chitosans (CT1-CT3) may be absorbed well from the gastrointestinal tract and inhibit neutrophil activation and oxidation of serum albumin that is frequently observed in patients plasma undergoing hemodialysis, resulting in a reduction in oxidative stress associated with uremia. PMID- 19559407 TI - Conformational properties of the disaccharide building units of hyaluronan. AB - The conformational space of the disaccharide building units of hyaluronan, beta (1-->4) and beta-(1-->3)-linked N-acetyl-beta-d-glucosamine (GN) and beta-D glucuronic acid (GA), has been investigated by density functional theory calculations at the B3LYP/6-31G(**) level. The study covered the anionic disaccharides, the neutral acids as well as the sodium salts in the isolated state and in aqueous solution using the PCM model approach. We elucidated the intramolecular hydrogen bonding interactions characterizing the most favoured conformers. The protonation and salt formation change these secondary interactions in the vicinity of the carboxyl group, resulting often in a considerable alteration of the conformational preferences. The Na(+) ion in the salt is involved in multiple bonding in the most stable structures: beyond the primary ionic bond with the carboxylate group it forms slightly weaker interactions with neighbouring oxygens. The main effect of protonation and salt formation on the electron density distribution is restricted to the surroundings of the broken/formed interactions near the carboxylate group. PMID- 19559406 TI - Control of cell motility by interaction of gangliosides, tetraspanins, and epidermal growth factor receptor in A431 versus KB epidermoid tumor cells. AB - Growth of epidermoid carcinoma cell lines, A431 and KB, has been known to be controlled by the interaction of epidermal growth factor (EGF) and its receptor (EGFR) with tyrosine kinase. Ganglioside GM3 was previously found to interact with EGFR and to inhibit EGFR tyrosine kinase. However, motility of these cells, controlled by EGFR and ganglioside, was not studied. The present study is focused on the control mechanism of the motility of these cells through interaction of ganglioside, tetraspanin (TSP), and EGFR. Key results are as follows: (i) The level of EGFR expressed in A431 cells is approximately 6 times higher than that expressed in KB cells, and motility of A431 cells is also much higher than that of KB cells, yet growth of A431 cells is either not affected or is inhibited by EGF. In contrast, growth of KB cells is enhanced by EGF. (ii) Levels of TSPs (CD9, CD82, and CD81) expressed in A431 cells are much higher than those expressed in KB cells, and TSPs expressed in A431 cells are reduced by treatment of cells with EtDO-P4, which inhibits the synthesis of glycosphingolipids (GSLs) and gangliosides. (iii) These TSPs are co-immunoprecipitated with EGFR in both A431 and KB cells, indicating that TSPs are closely associated with EGFR. (iv) High motility of A431 cells is greatly reduced, while low motility of KB cells is not affected, by treatment of cells with EtDO-P4. These results, taken together, suggest that there is a close correlation between high motility of A431 cells and high expression of EGFR and TSPs, and between ganglioside GM3/GM2 and TSP. A similar correlation was suggested between the low motility of KB cells and low levels of EGFR and TSP. The correlation between high motility and high level of EGFR with the ganglioside-TSP complex in A431 cells is unique. This is in contrast to our previous studies that indicate that motility of many types of tumor cells is inhibited by a high level of CD9 or CD82, together with growth factor receptors and integrins. PMID- 19559409 TI - Darwin and the ghost of Phineas Gage: neuro-evolution and the social brain. PMID- 19559408 TI - TLR4-independent and PKR-dependent interleukin 1 receptor antagonist expression upon LPS stimulation. AB - Dendritic cells (DCs) induce innate immune responses by recognizing bacterial LPS through TLR4 receptor complexes. In this study, we compared gene expression profiles of TLR4 knockout (TLR4(neg)) DCs and wild type (TLR4(pos)) DCs after stimulating with LPS. We found that the expression of various inflammatory genes by LPS were TLR4-independent. Among them, interleukin 1 receptor antagonist (IL 1rn) was of particular interest since IL-1rn is a potent natural inhibitor of proinflammatory IL-1. Using RT-PCR, real-time PCR, immunoblotting and ELISA, we demonstrated that IL-1rn was induced by DCs stimulated with LPS in the absence of TLR4. 2-Aminopurine, a pharmacological PKR inhibitor, completely abrogated LPS induced expression of IL-1rn in TLR4(neg) DCs, suggesting that LPS-induced TLR4 independent expression of IL-1rn might be mediated by PKR pathways. Considering that IL-1rn is a physiological inhibitor of IL-1, TLR4-independent and PKR dependent pathways might be crucial in counter-balancing proinflammatory effector functions of DCs resulted from TLR4-dependent activation by LPS. PMID- 19559411 TI - Effects of environmental noise exposure on ambulatory blood pressure in young adults. AB - Epidemiological studies have demonstrated that environmental noise exposure is associated with hypertension in middle-aged and older populations, but the relationship in the young subpopulation and between the genders is still unclear. This panel study investigated effects of environmental noise exposure on 24-h ambulatory blood pressure in 60 adults aged 18-32 years. Individual noise exposure and personal blood pressure were measured simultaneously for 30 males and 30 females. Linear mixed-effects regression models were applied to estimate effects. Total subjects (56.6+/-16.5A-weighted decibels (dBA)) had transient elevations of 1.15 (95% CI=0.86-1.43)mmHg SBP and 1.16 (0.93-1.38)mmHg DBP at daytime, as well as 0.74 (0.21-1.26)mmHg SBP and 0.77 (0.34-1.20)mmHg DBP at nighttime, significantly associated with a 5-dBA increase in noise exposure. Such effects on SBP and DBP still persisted at the 30- and 60-min time-lagged noise exposure. Per 5-dBA increase in 24-h average noise exposure was significantly associated with sustained increments of 1.15 (0.76-1.54)mmHg SBP and 1.27 (0.96 1.58)mmHg DBP in males (57.4+/-16.0dBA), as well as the higher levels of 1.65 (1.36-1.94)mmHg SBP and 1.51 (1.27-1.75)mmHg DBP in females (55.9+/-17.0dBA). We found that environmental noise exposure may have elevated effects on adults' blood pressure. Young females are more susceptible to noise exposure than males. PMID- 19559412 TI - Clinical assessment of fracture risk and novel therapeutic strategies to combat osteoporosis. AB - OBJECTIVE: To review the latest tools in the clinical assessment of fracture risk and to review new and emerging options for osteoporosis therapy. DESIGN: Retrospective analysis of published studies regarding the diagnosis and treatment of osteoporosis. RESULT(S): Large-scale epidemiologic data were recently assembled by the World Health Organization to produce a Web-based clinical assessment tool, FRAX, which uses clinical and historical data to provide prompt assessment and quantitation of fracture risk. The FRAX models were developed from studying population-based cohorts in Europe, North America, Asia, and Australia. The FRAX algorithms indicate the 10-year probability of hip fracture and the 10 year probability of a major osteoporotic fracture (at the clinical spine, forearm, hip, or shoulder) on which to base treatment decisions. Recent progress in the study of bone metabolism including anabolic pathways that enhance bone maintenance, is anticipated to improve the ways in which skeletal health can be maintained and osteoporosis can be treated. CONCLUSION(S): Using FRAX, fracture risk in now easily assessed in the clinical setting. New and emerging treatment strategies for bone maintenance are reviewed. Improved assessment of fracture risk, combined with tailored therapies for at-risk patients, will increase the number of patients who receive appropriate bone-sparing therapies. PMID- 19559410 TI - Electrophysiological evidence for aging effects on local contextual processing. AB - We used event-related potentials to investigate how aging affects local contextual processing. Local context was defined as the occurrence of a short predictive series of visual stimuli before delivery of a target event. Stimuli were presented to either the left or right visual field and consisted of 15% targets (downward facing triangle) and 85% of equal numbers of three types of standards (triangles facing left, upwards and right). Recording blocks consisted of targets preceded by either randomized sequences of standards or by sequences including a three-standard predictive sequence signaling the occurrence of a subsequent target event. Subjects pressed a button in response to targets. Predictive local context affected target detection by reducing the duration of stimulus evaluation compared to detection of non-predictive random targets comparably for both young and older adults, as shown by a P3b latency shift. The peak of an earlier latency context positivity, which was seen only in the predicted compared to the random target condition, was prolonged in the older population compared to young adults. Finally, older subjects elicited a late sustained positivity in the predictive condition, not seen in the younger subjects. Taken together, these findings suggest that local contextual effects on target detection processes are altered with age. PMID- 19559413 TI - Virtual hysterosalpingography and hysteroscopy: assessment of uterine cavity and fallopian tubes using 64-detector computed tomography data sets. AB - Hysterosalpingography is the primary technique in providing coarse information on the morphology of endometrial cavity and fallopian tubes. In this preliminary study, 64-detector computed tomography was used for three-dimensional imaging of endometrium and fallopian tubes. PMID- 19559414 TI - Spontaneous migration of a prosthetic mesocaval shunt to the duodenum: endoscopic diagnosis of an unusual complication of shunt surgery. PMID- 19559415 TI - The big burden of obesity. PMID- 19559416 TI - Natural orifice transluminal endoscopic surgery gastrotomy closure with an over the-endoscope clip: a randomized, controlled porcine study (with videos). AB - BACKGROUND: Secure endoscopic closure of transgastric natural orifice transluminal endoscopic surgery (NOTES) access is of paramount importance. The over-the-scope clip (OTSC) system has previously been shown to be effective for NOTES gastrotomy closure. OBJECTIVE: To compare OTSC gastrotomy closure with surgical closure. DESIGN: Randomized, controlled animal study. SETTING: Animal facility laboratory. ANIMALS: Thirty-six female domestic pigs. INTERVENTIONS: Gastrotomies were created by using a needle-knife and an 18-mm balloon. The animals were subsequently randomized to either open surgical repair with interrupted sutures or endoscopic repair with 12-mm OTSCs. In addition, pressurized leak tests were performed in ex vivo specimens of 18-mm scalpel incisions closed with suture (n = 14) and of intact stomachs (n = 10). MAIN OUTCOME MEASUREMENTS: The mean time for endoscopic closure was 9.8 minutes (range 3-22, SD 5.5). No complications occurred during either type of gastrotomy closure. At necropsy, examination of all OTSC and surgical closures demonstrated complete sealing of gastrotomy sites without evidence of injury to adjacent organs. Pressurized leak tests showed a mean burst pressure of 83 mm Hg (range 30 140, SD 27) for OTSC closures and 67 mm Hg (range 30-130, SD 27.7) for surgical sutures. Ex vivo hand-sewn sutures of 18-mm gastrotomies (n = 14) exhibited a mean burst pressure of 65 mm Hg (range 20-140, SD 31) and intact ex vivo stomachs (n = 10) had a mean burst pressure of 126 mm Hg (range 90-170, SD 28). The burst pressure of ex vivo intact stomachs was significantly higher compared with OTSC closures (P < .01), in vivo surgical closures (P < .01), and ex vivo hand-sewn closures (P < .01). There was a trend toward higher burst pressures in the OTSC closures compared with surgical closures (P = .063) and ex vivo hand-sewn closures (P = .094). In vivo surgical closures demonstrated similar burst pressures compared with ex vivo hand-sewn closures (P = .848). LIMITATIONS: Nonsurvival setting. CONCLUSION: Endoscopic closure by using the OTSC system is comparable to surgical closure in a nonsurvival porcine model. This technique is easy to perform and is suitable for NOTES gastrotomy closure. PMID- 19559417 TI - Esophageal intramural pseudodiverticula with endoscopic and radiographic demonstration of intramural trackings. PMID- 19559418 TI - Endoscopic transpapillary gallbladder drainage with the SpyGlass cholangiopancreatoscopy system. PMID- 19559420 TI - Endoscopic removal of a fishhook in the esophagus. PMID- 19559419 TI - Confocal laser endomicroscopy in Barrett's esophagus and endoscopically inapparent Barrett's neoplasia: a prospective, randomized, double-blind, controlled, crossover trial. AB - BACKGROUND: The detection of high-grade dysplasia and cancer in Barrett's esophagus (BE) can be challenging. Confocal laser endomicroscopy (CLE) allows in vivo visualization of mucosal histology during endoscopy. OBJECTIVE: To determine whether CLE with optical biopsy and targeted mucosal biopsy improves the diagnostic yield of endoscopically inapparent, BE-associated neoplasia compared to standard endoscopy with a 4-quadrant, random biopsy protocol. DESIGN: Prospective, double-blind, randomized, crossover study. SETTING: Single, tertiary care academic center. PATIENTS: This study involved patients with BE undergoing routine surveillance or referred for treatment of nonlocalized, endoscopically inapparent, BE-associated neoplasia. INTERVENTION: All participants underwent both a confocal endomicroscopy with a targeted biopsy procedure and standard endoscopy with a 4-quadrant biopsy procedure in a randomized order. MAIN OUTCOME MEASUREMENTS: Increase in diagnostic yield for neoplasia, reduction in mucosal biopsy number, final pathologic diagnosis. RESULTS: CLE with targeted biopsy almost doubled the diagnostic yield for neoplasia and was equivalent to the standard protocol for the final diagnosis of neoplasia. Two thirds of patients in the surveillance group did not need any mucosal biopsies at all. LIMITATION: Single-center study. CONCLUSION: CLE with targeted biopsy significantly improves the diagnostic yield for endoscopically inapparent BE neoplasia compared to a standard endoscopy with a random-biopsy protocol. CLE with targeted biopsy also greatly reduces the number of biopsies needed per patient and allows some patients without neoplasia to completely forgo mucosal biopsy. PMID- 19559421 TI - Fasciola hepatica causing acute pancreatitis complicated by biliary sepsis. PMID- 19559422 TI - Large solitary rectal ulcer as a complication of prostate brachytherapy. PMID- 19559423 TI - An unusual presentation of eosinophilic esophagitis. PMID- 19559424 TI - Massive hemorrhage from a pulmonary vein-esophageal fistula: a late complication of Histoacryl glue injection. PMID- 19559425 TI - Endoscopic cyanoacrylate versus transjugular intrahepatic portosystemic shunt for gastric variceal bleeding: a single-center U.S. analysis. AB - BACKGROUND AND OBJECTIVES: Gastric variceal hemorrhage treatment remains a difficult issue for clinicians. There is controversy regarding whether first-line treatment should be endoscopic therapy with cyanoacrylate glue or placement of a transjugular intrahepatic portosystemic shunt (TIPS). We compared these methods on the basis of rebleeding, survival, and complications. DESIGN, SETTING, PATIENTS, AND INTERVENTIONS: This was a retrospective cohort analysis of cirrhotic patients with gastric variceal hemorrhage treated with endoscopic cyanoacrylate therapy or TIPS placement at a single U.S. center from 1997 to 2007. The groups were compared for rebleeding at 72 hours, 3 months, and 1 year; survival rates at 3 months and 1 year; and acute and extended complications and morbidity. MAIN OUTCOME MEASUREMENTS AND RESULTS: A total of 105 patients were included. There were no significant pretreatment differences between the 2 groups in age, sex, MELD (Model for End-Stage Liver Disease) score at the time of admission, or cause of liver disease. There were no significant differences in rebleeding at 72 hours, 3 months, and 1 year; survival at 3 months and 1 year; and aggregate long-term survival or acute complications. However, the TIPS group had a higher rate of long-term morbidity requiring hospitalization (41% with a TIPS and 1.6% in the cyanoacrylate arm, P < .0001). LIMITATIONS: Retrospective and uncontrolled samples. CONCLUSION: In patients with similar characteristics, cyanoacrylate therapy performed as well as a TIPS in controlling and preventing gastric variceal hemorrhage with no significant differences in survival. Patients receiving cyanoacrylate therapy experienced significantly less long-term morbidity related to therapy than patients who received a TIPS. Cyanoacrylate therapy appears to be safe and effective and compares favorably with TIPS therapy. PMID- 19559426 TI - Confocal laser endomicroscopy. PMID- 19559427 TI - A randomized trial of endoscopic cyanoacrylate injection for acute gastric variceal bleeding: 0.5 mL versus 1.0 mL. AB - BACKGROUND: Endoscopic injection of N-butyl-2-cyanoacrylate is the preferred method to treat acute gastric variceal bleeding (GVB). However, its rebleeding rate remains high. OBJECTIVE: To compare an injection containing 0.5 mL of cyanoacrylate (group A) with an injection containing 1.0 mL of cyanoacrylate (group B). DESIGN: A single-center, randomized, controlled trial. SETTING: A tertiary referral center. MAIN OUTCOME MEASUREMENT: Occurrence of rebleeding. PATIENTS: Patients with acute gastric variceal bleeding. RESULTS: Forty-four patients in group A and 47 patients in group B were studied; their clinical characteristics were similar. The treatment stopped active bleeding in approximately 90% of cases in both groups. The rebleeding rate was 29.8% (14/47) in group B compared with 38.6% (17/44) in group A (P = .504; 95% CI, -10.592 to 28.280). On multivariate analysis, concomitant hepatocellular carcinoma, infection, and the size of the gastric varices were independent determinants of rebleeding. More patients in group B than in group A had postinjection fever (>37.5 degrees C) (23/47 vs 12/44, P = .059). Treatment failure, complications, 30-day mortality, and survival did not differ between the 2 groups. CONCLUSIONS: Due to the small number of study patients, a double dose of cyanoacrylate injection for GVB cannot be proven to have better hemostatic efficacy than a single dose. Multicenter studies with larger patient numbers are necessary to determine whether a double dose is in fact more efficacious. PMID- 19559429 TI - Successful endoscopic management of a bleeding ulcer in a giant epiphrenic diverticulum. PMID- 19559428 TI - An open-label, prospective trial of cryospray ablation for Barrett's esophagus high-grade dysplasia and early esophageal cancer in high-risk patients. AB - BACKGROUND: Endoscopic ablation of Barrett's esophagus (BE) is a treatment option for patients with high-grade dysplasia (HGD) and intramucosal carcinoma (IMCA). OBJECTIVE: To assess the safety and efficacy of a unique noncontact method of liquid nitrogen cryoablation as measured by histologic response rate and cancer free survival. DESIGN: Single-center, nonrandomized cohort study. SETTING: Referral center, conducted between September 2005 and September 2008. PATIENTS: Patients with BE and HGD or IMCA who were deemed inoperable or who refused esophagectomy. Age, length of BE, and previous ablation were not exclusion criteria. INTERVENTION: Cryoablation every 6 weeks until endoscopic resolution. EMR was used for pathologic staging of nodular areas before cryoablation and focal residual areas during the follow-up period. MAIN OUTCOME MEASUREMENTS: Histologic response was defined by the worst pathology obtained at any level of the esophagus or gastric cardia in 1 of 3 categories: (1) incremental = absence of HGD and IMCA in all biopsy specimens, (2) partial = residual IMCA with absence of any dysplasia, and (3) complete = absence of any intestinal metaplasia or dysplasia. RESULTS: Thirty patients underwent ablation; 9 had undergone previous ablation or mucosectomy. Twenty-seven of 30 patients (90%) had downgrading of pathology stage after treatment. Elimination of cancer or downgrading of HGD at last follow-up was 68% for HGD and 80.0% for IMCA, with a median follow-up period of 12 months (25th percentile, 6; 75th percentile, 24). Minor adverse events included mild pain (n = 7), a low incidence of mild strictures (n = 3), and lip ulcer (n = 1). One major adverse event (perforation) in a patient with Marfan syndrome occurred with the prototype system. During follow-up, 3 of 6 patients with complete response had recurrence of dysplasia or cancer in the gastric cardia. LIMITATIONS: A nonrandomized, single-center study with a heterogeneous cohort of patients. CONCLUSIONS: Patients with BE and HGD or IMCA have a positive response to endoscopic cryotherapy at 1-year follow-up. PMID- 19559430 TI - Liver injury from endoscopic insertion of self-expandable metallic stent to relieve biliary obstruction: a fatal complication. PMID- 19559431 TI - First successful antegrade single-balloon enteroscopy in a 3-year-old with occult GI bleeding. AB - BACKGROUND: Balloon enteroscopy is an emerging technique to allow access to the small intestine for both diagnostic and therapeutic purposes. To date, there have been few published data documenting the safety and efficacy of balloon enteroscopy in small children. OBJECTIVE: To describe our experience with single balloon enteroscopy (SBE) in a 37-month-old toddler with occult GI bleeding. DESIGN: A single case report. SETTING: A free-standing, academic children's hospital in Denver, Colorado. PATIENT: The patient was a 37-month-old, 13.5-kg toddler with persistent heme-positive stools, severe microcytic anemia, and hypoalbuminemia. Previous workup was significant for eosinophilic inflammation in the antrum and a video capsule study showing erythematous lesions in the small bowel. INTERVENTION: An antegrade SBE was performed with the child under general endotracheal anesthesia, with biopsy specimens obtained from identified lesions in the jejunum and ileum. MAIN OUTCOME MEASUREMENTS: Complications and successful treatment of symptoms were the primary endpoints. RESULTS: The procedure was performed successfully in 85 minutes, passing an estimated 200 cm beyond the pylorus, without complications. Identification of the lesions as consistent with eosinophilic enteropathy led to successful treatment with an elimination diet and corticosteroids. LIMITATIONS: The primary limitation of this study is that it is a single case report. Therefore, it is difficult to make a generalized statement regarding the safety and efficacy of balloon enteroscopy in toddlers of this size. CONCLUSIONS: Antegrade SBE can be a well-tolerated and effective procedure to evaluate occult GI bleeding in children as young as 3 years of age. Further study is needed to better establish safety parameters for balloon enteroscopy in small pediatric patients. PMID- 19559432 TI - Diagnostic and therapeutic procedure with a short double-balloon enteroscope and cholangioscopy in a patient with acute cholangitis due to hepatolithiasis. PMID- 19559433 TI - Simulator training improves practical skills in therapeutic GI endoscopy: results from a randomized, blinded, controlled study. AB - BACKGROUND: Therapeutic endoscopic procedures are technically challenging and have higher complication rates than diagnostic procedures. Less-experienced practitioners are significantly more likely to have a complication, yet there is very limited evidence of the efficacy of training in such procedures. OBJECTIVE: To evaluate the effect of knowledge-based teaching and hands-on, simulation-based skills training in 4 therapeutic endoscopic procedures: control of nonvariceal upper GI bleeding, polypectomy, stricture dilation, and percutaneous endoscopic gastrostomy (PEG) tube insertion. DESIGN: Single-blind, randomized, controlled trial. SETTING: Nationally accredited therapeutic endoscopy course. PARTICIPANTS AND INTERVENTION: Participants were randomized into groups to assess the effect of knowledge-based teaching and into subjects or controls to assess skills-based training. All delegates had an initial knowledge and skills assessment by blinded experts. Subjects received one-half day of hands-on skills training, whereas the controls did not. All delegates were retested on their procedural skills. MAIN OUTCOME MEASUREMENTS: Knowledge was assessed by using a multiple-choice questionnaire. Practical skills were assessed using station-specific checklists and a global score. RESULTS: Twenty-eight participants were randomized. There was no significant effect of knowledge-based teaching on the participants' practical skills or initial multiple-choice questionnaire score, although there was a trend toward improvement. There was a significant improvement in the subjects compared with the controls in the performance of polypectomy, control of upper GI bleeding, and esophageal dilation. There were no significant differences for PEG tube insertion. LIMITATIONS: The 2-person nature of PEG tube insertion may have resulted in performance improvement despite a lack of training. CONCLUSION: Hands on skills training significantly improved performance in 3 therapeutic modalities. Knowledge-based teaching alone did not have a measurable effect. These results strongly support the benefit of intensive hands-on, simulation based courses for endoscopic skills training. PMID- 19559434 TI - More aggravated osteoporosis in lateral trochanter compared to femoral neck with age: contributing age difference between inter-trochanteric and femoral neck fractures in elderly patients. AB - In the medical literature, elderly patients with inter-trochanteric fractures are considered older than those with femoral neck fractures, but the reason for this age-related difference remains unclear. Inconsistent bone mass re-distribution with age in both regions was hypothesised and the change in bone mineral density (BMD) was studied. This study aims to investigate another possible pathomechanism of reducing bone strength in the proximal femur with increasing age. The findings might help in developing a better method for preventing osteoporotic hip fractures. PATIENTS AND METHODS: In the BMD study, 400 consecutive female patients were divided into eight groups, comprising 50 individuals each. Except for the first group (16-20 years), all other groups were divided by 10-year age increments up to 90 years. Relative BMD change with age in different regions in each group was compared. RESULTS: The BMD ratio of the lateral trochanter to the neck gradually reduced with age, whereas that of the medial trochanter to the neck gradually increased. The comparisons were statistically significant (p<0.001 in both cases). CONCLUSION: One important predisposing factor to affect patients with inter-trochanteric fractures older than those with femoral neck fractures may be that the BMD is further lowered in the lateral trochanter compared to the femoral neck with age. It may be caused by lack of compressive loads in the lateral trochanter in daily activity. All devices which may increase bone mass in the lateral trochanter, theoretically, may be helpful in lowering the incidence of hip fractures. PMID- 19559435 TI - Nurses' perceptions of informed consent and their related roles in Korea: an exploratory study. AB - BACKGROUND: Informed consent is based on the Western notion of autonomy that patients have the right to know about their illnesses and make decisions about their health. However, there may be difficulty in accepting informed consent in Eastern cultures based on Confucianism, such as Korea. Even though nurses have no legal binds to informed consent, they have participated in the process of obtaining informed consent as an administrative assistant in Korea and then have had moral distress. OBJECTIVES: The purpose of this study was to investigate nurses' perception of informed consent and their role in establishing a better process of informed consent in Korea. METHODS: This exploratory study employed in depth interviews with nurses. Participants were 12 nurses from selected departments (orthopedics surgery, cardiac surgery, and obstetrics & gynecology) from two university hospitals in Seoul and Kwang-ju, Korea. A semi-structured interview guide was constructed from the literature and used to elicit responses. RESULTS: Content analysis showed that nurses perceived a lot of problems in the process of obtaining informed consent and noted the need to modify this process. Although the role of nurses was found to be insignificant in the process of obtaining informed consent, some nurses wanted to participate in the process in order to advocate for patients and to verify if patients had received the information they needed. CONCLUSIONS: Nurses can take a proactive role in the process of informed consent, as advocator as well as witness. This may be influential in balancing patient autonomy with paternalism, changing the skewed patient-physician relationship to one of mutual respect. PMID- 19559437 TI - Finite element simulation of interactions between pelvic organs: predictive model of the prostate motion in the context of radiotherapy. AB - The setting up of predictive models of the pelvic organ motion and deformation may prove an efficient tool in the framework of prostate cancer radiotherapy, in order to deliver doses more accurately and efficiently to the clinical target volume (CTV). A finite element (FE) model of the prostate, rectum and bladder motion has been developed, investigating more specifically the influence of the rectum and bladder repletions on the gland motion. The required organ geometries are obtained after processing the computed tomography (CT) images, using specific softwares. Due to their structural characteristics, a 3D shell discretization is adopted for the rectum and the bladder, whereas a volume discretization is adopted for the prostate. As for the mechanical behavior modelling, first order Ogden hyperelastic constitutive laws for both the rectum and bladder are identified. The prostate is comparatively considered as more rigid and is accordingly modelled as an elastic tissue undergoing small strains. A FE model is then created, accounting for boundary and contact conditions, internal and applied loadings being selected as close as possible to available anatomic data. The order of magnitude of the prostate motion predicted by the FE simulations is similar to the measurements done on a deceased person, accounting for the delineation errors, with a relative error around 8%. Differences are essentially due to uncertainties in the constitutive parameters, pointing towards the need for the setting up of direct measurement of the organs mechanical behavior. PMID- 19559438 TI - Morphology and fracture of enamel. AB - This study examines the inter-relation between enamel morphology and crack resistance by sectioning extracted human molars after loading to fracture. Cracks appear to initiate from tufts, hypocalcified defects at the enamel-dentin junction, and grow longitudinally around the enamel coat to produce failure. Microindentation corner cracks placed next to the tufts in the sections deflect along the tuft interfaces and occasionally penetrate into the adjacent enamel. Although they constitute weak interfaces, the tufts are nevertheless filled with organic matter, and appear to be stabilized against easy extension by self healing, as well as by mutual stress-shielding and decussation, accounting at least in part for the capacity of tooth enamel to survive high functional forces. PMID- 19559439 TI - Simultaneous determination of dimethylamine, trimethylamine and trimethylamine-n oxide in aquatic products extracts by ion chromatography with non-suppressed conductivity detection. AB - An ion chromatography method with non-suppressed conductivity detection was developed for the simultaneous determination of dimethylamine (DMA), trimethylamine (TMA) and trimethylamine-n-oxide (TMAO) in aquatic products. They were separated by means of cation-exchange chromatography using a 3.0 mmol/L methanesulfonic acid solution as eluent and an IonPac CS17 column (250 mm x 4 mm i.d.) as the separation column. Detection limits of dimethylamine, trimethylamine and trimethylamine-n-oxide were 0.06, 0.08 and 0.10 mg/L, respectively. The relative standard deviations (RSDs) of peak area were less than 3.53%. The recoveries were between 93.7% and 104.1%. Unlike traditional methods, this validated method is inexpensive and stable. PMID- 19559440 TI - Using gold nanoparticles to improve the recovery and the limits of detection for the analysis of monohydroxy-polycyclic aromatic hydrocarbons in urine samples. AB - We present a novel approach to improve the analytical figures of merit of solid phase extraction high-performance liquid chromatography (SPE-HPLC) for the analysis of monohydroxy-polycyclic aromatic hydrocarbons in urine samples. The novel alternative substitutes the evaporation step that is currently used in SPE HPLC methodology with a pre-concentration procedure that extracts metabolites with gold nanoparticles. The analytical potential of the new approach is evaluated with the following six metabolites: 9-hydroxyphenanthrene, 2 hydroxyfluorene, 1-hydroxypyrene, 6-hydroxychrysene, 3-hydroxybenzo[a]pyrene and 4-hydroxybenzo[a]pyrene. We demonstrate that the substitution of the evaporation step with the gold nanoparticles procedure improves the overall recoveries, the relative standard deviations of the average recoveries and the limits of detection of SPE-HPLC analysis. The overall recoveries of the studied metabolites varied from 59.7 +/- 3.6% (2-hydroxyfluorene) to 92.3 +/- 2.5% (6 hydroxychrysene). The relative standard deviations of the average recoveries were lower than 6%. The limits of detection were at the parts-per-trillion levels and varied from approximately 2 pg mL(-1) (6-hydroxychrysene) to approximately 18 pg mL(-1) (2-hydroxyfluorene). PMID- 19559441 TI - Analysis by desorption of volatile impurities from an ionic liquid solution in an unmodified gas chromatograph inlet. AB - A procedure for using ionic liquids to determine volatile impurities in compounds or matrices that are soluble in an ionic liquid is described. Using a conventional autosampler a droplet of ionic liquid solution is suspended in the inlet of the gas chromatograph and analytes are desorbed onto a GC column using splitless injection conditions. Results are presented for 1,2-propanediol, nonane, N,N-dimethylacetamide, and mesitylene in two different compounds in the ionic liquids trihexyltetradecylphosphonium bis(trifluoromethylsulfonyl)imide and trihexyltetradecylphosphonium dicyanamide. PMID- 19559442 TI - Magnetically immobilized frits for the preparation of packed columns used in capillary electrochromatography. AB - Fabrication of porous frits to retain stationary phases is a critical issue in column preparation for capillary electrochromatography (CEC). In this work, porous frits were prepared by applying an external magnetic field to magnetically responsive particles placed inside a fused-silica capillary. Three batches of uniform magnetite spheres with particle diameters of 0.3, 0.4, and 0.6 microm and saturation magnetization values of 73.03, 74.41, and 77.83 emu/g, respectively, were used as frit particles and octadecyl- and phenyl-bonded silica gels were packed successfully into frit-containing capillaries. The performance of the resulting magnetically immobilized frits and packed columns was evaluated. The electroosmotic mobilities in capillaries containing outlet frit only were found to be reduced by 2-4% whereas the plate heights of an unretained marker increased by 30-50% as compared to those in open capillaries. These variations are believed to be associated with the inhomogeneities of the packed structure of the frits. The magnetically immobilized frits showed adequate mechanical strength to withstand the flow drag force, allowing separation in capillaries packed with 5 mum stationary phases up to 10-15 cm, thus rendering column efficiency and reproducibility comparable with those obtained with sintered frits. Taken together, retaining frits made of uniform magnetite particles serves as a viable alternative to sintered frits for column preparation, which offers several distinct advantages such as ease of preparation, improved durability as compared to sintered frits where the removal of the polyimide coating makes the packed column susceptible to breakage, and use of large-bore capillaries for semipreparative separations. PMID- 19559443 TI - Determination of methotrexate in human serum by high-performance liquid chromatography combined with pseudo template molecularly imprinted polymer. AB - A pseudo template molecularly imprinted polymer (MIP) was prepared for methotrexate (MTX) and a RP-HPLC method combined with the MIP was developed for the determination of MTX in human serum. Because of the poor solubility of MTX in common MIP preparation solvents, trimethoprim (TMP), a molecule having the similar imprinting sites as MTX, is selected as the pseudo template. The MIP was prepared using methacrylic acid (MAA) and ethylene glycol dimethacrylate as functional monomer and cross-linker, respectively. 1H NMR study showed highly strong interaction between TMP and MAA with hydrogen bonds. Chromatographic behaviors indicated that the TMP-MIP possessed excellent affinity and selectivity for MTX. And the imprinting factor for MTX was high up to 9.5 when 7:3 of acetonitrile:methanol (v/v) was used as mobile phase. Moreover, TMP-MIP was used as the solid-phase extraction (SPE) material to enrich the target compound MTX in human serum samples for HPLC analysis. The SPE process was carefully optimized and good recoveries of MTX were obtained as 81.6-86.2% with RSD of 0.22-1.84% when the spiked concentration of MTX was 2.0-10.0 microg mL(-1) in human serum samples. The results indicated that the pseudo template MIP can be applied to preconcentration, purification and analysis of MTX in clinic samples. PMID- 19559444 TI - Use of an injection port for thermochemolysis-gas chromatography/mass spectrometry: rapid profiling of biomaterials. AB - A simple and direct approach was developed for thermochemolytic analysis of a wide range of biomolecules present in plant materials using an injection port of a gas chromatograph/mass spectrometer (GC/MS) and a novel solids injector consisting of a coiled stainless steel wire placed inside a modified needle syringe. Optimum thermochemolysis (or Thermally Assisted Hydrolysis/Methylation) was achieved by using a suitable methanolic solution of trimethylsulfonium hydroxide (TMSH) or tetramethylammonium hydroxide (TMAH) with an injection port temperature of 350 degrees C. Intact, methylated flavonoids, saccharides, phenolic and fatty acids, lignin dimers and diterpene resin acids were identified. Samples include tea leaves, hemicelluloses, lignin isolates and herbal medicines. Unexpected chromatographic results using TMAH reagent revealed the presence of intact methylated trisaccharides (658 Da) and structurally informative dimer lignin markers. PMID- 19559445 TI - Menarche and assessment of body mass index in adolescent girls. AB - OBJECTIVES: To determine whether menarcheal status in individual girls biases assessments of overweight and obesity when using body mass index (BMI) criteria from age-based reference data. STUDY DESIGN: Data were analyzed for 2145 girls aged 8.00 to 16.99 years was examined from 1999 to 2002 as part of the National Health and Nutrition Examination Survey (NHANES). RESULTS: An estimate of the menarche-related effect on BMI assessment at each age was calculated as K = (m M)beta, in which m = the menarcheal status of a girl (0,1); M = the prevalence of menarcheal status; and beta = the regression coefficient of BMI on m. At levels of BMI near the percentile cutoff points defining overweight and obesity (85th, 95th) the prevalence of menarcheal status is higher and the BMI differences between menarcheal and premenarcheal girls (beta) are smaller than those observed when all girls are considered. The average effects on BMI of menarche that occurs earlier than the population mean age or of menarche that occurs later than the population mean age generally are 0.05). In conclusion, this study failed to demonstrate beneficial effects of a postinsemination treatment of primiparous and multiparous cows with cloprostenol on ovulation and PR in a typical dairy cattle operation in East Germany. PMID- 19559472 TI - Strategies to improve fertility in Bos indicus postpubertal heifers and nonlactating cows submitted to fixed-time artificial insemination. AB - Two experiments were designed to evaluate strategies to increase fertility of Bos indicus postpubertal heifers and nonlactating cows submitted to a fixed-time artificial insemination (TAI) protocol consisting of an intravaginal device containing 1.9 g of progesterone (CIDR) insertion+estradiol benzoate on Day 0, CIDR withdrawal+estradiol cypionate on Day 9, and TAI on Day 11. In Experiment 1, heifers (n=1153) received a new or an 18-d previously used CIDR and, on Day 9, prostaglandin F(2 alpha) (PGF(2 alpha))+0, 200, or 300 IU equine chorionic gonadotropin (eCG). Heifers treated with a new CIDR had greater (least squares means+/-SEM) serum concentration of progesterone on Day 9 (3.06+/-0.09 ng/mL vs. 2.53+/-0.09 ng/mL; P<0.05) and a smaller follicle at TAI (11.61+/-0.11 mm vs. 12.05+/-0.12 mm; P<0.05). Heifers with smaller follicles at TAI had lesser serum progesterone concentrations on Day 18 and reduced rates of ovulation, conception, and pregnancy (P<0.05). Treatment with eCG improved (P<0.05) follicle diameter at TAI (11.50+/-0.10mm, 11.90+/-0.11 mm, and 12.00+/-0.10mm for 0, 100, and 200 IU, respectively), serum progesterone concentration on Day 18 (2.77+/-0.11 ng/mL, 3.81+/-0.11 ng/mL, and 4.87+/-0.11 ng/mL), and rates of ovulation (83.8%, 88.5%, and 94.3%) and pregnancy (41.3%, 47.0%, and 46.7%). In Experiment 2, nonlactating Nelore cows (n=702) received PGF(2 alpha) treatment on Days 7 or 9 and, on Day 9, 0 or 300 IU eCG. Cows receiving PGF(2 alpha) on Day 7 had lesser serum progesterone concentrations on Day 9 (3.05+/-0.21 ng/mL vs. 4.58+/-0.21 ng/mL; P<0.05), a larger follicle at TAI (11.54+/-0.21 mm vs. 10.84+/-0.21 mm; P<0.05), and improved (P<0.05) rates of ovulation (85.4% vs. 77.0%), conception (60.9% vs. 47.2%), and pregnancy (52.0% vs. 36.4%). Treatment with eCG improved (P<0.05) serum progesterone concentration on Day 18 (3.24+/-0.14 ng/mL vs. 4.55+/-0.14 ng/mL) and the rates of ovulation (72.4% vs. 90.0%) and pregnancy (37.5% vs. 50.8%). In conclusion, giving PGF(2 alpha) earlier in the protocol in nonlactating cows and eCG treatment in postpubertal heifers and nonlactating cows improved fertility in response to a TAI (progesterone+estradiol) protocol. PMID- 19559473 TI - Ovarian cysts in high-yielding dairy cows. AB - We examined the hormonal and morphologic changes associated with ovarian cyst formation in high-yielding dairy cows. Follicle fluid was aspirated from 90 cysts and 15 preovulatory and 18 subordinate follicles and used for hormonal determination. Pieces of cystic wall were subjected to morphologic and immunohistochemical evaluation. Cysts were characterized by low concentrations of insulin, insulin-like growth factor-I (IGF-I), and glucose and high activity of IGF binding proteins (IGFBPs). Insulin and IGF-I levels were (mean+/-SEM) 205+/ 22 pg/mL and 146+/-42 ng/mL in preovulatory follicles and 3+/-1 pg/mL and 61+/-6 ng/mL in cysts, respectively (P<0.001). Insulin-like growth factor-binding proteins activity was about 10 times higher in cysts than in preovulatory follicles. Cysts were classified into three types according to their estradiol-to progesterone (E/P) ratio. Type 1 cysts (n=23) exhibited the highest E/P ratio (10.8+/-2.3), partial loss of granulosa cells, and severe morphologic changes in the theca interna. Expression of P(450) side-chain cleavage and P(450) 17 alpha hydroxylase was noted in theca cells and expression of inhibin-alpha in granulosa cells. Type 2 cysts (n=35) had a low E/P ratio (0.07+/-0.02), and patches of luteal-like tissue in the cystic wall. Type 3 cysts (n=32) had an E/P ratio of 0.91+/-0.17, and no recognizable granulosa or theca cells. In summary, intrafollicular steroid levels as expressed by E/P ratio, together with IGF-I and insulin levels and morphologic changes in the follicular wall, may serve as accurate cyst-classification parameters. Because IGF-I and/or insulin play an essential role in the final stage of follicle development, it can be speculated that abnormal levels of these metabolic hormones might lead to follicle dysfunction, resulting in follicular regression or cyst formation. PMID- 19559474 TI - Effect of recombinant gonadotropins on embryo quality in superovulated rabbit does and immune response after repeated treatments. AB - This study aimed first to evaluate the effect of recombinant human FSH (rhFSH) with and without recombinant human LH (rhLH) on fresh and frozen-thawed embryo development and also to analyze the immune response of rabbit does (Oryctolagus cuniculus) subjected to repeated rhFSH treatments. Nulliparous New Zealand White does were used. In Experiment 1, 120 does were superovulated with 25 IU rhFSH alone or in combination with 5% or 10% rhLH (1.25 IU or 2.50 IU rhLH). A total of 1116 embryos at the compacted morula stage were cultured at 38.5 degrees C, 5% CO(2), and saturated humidity for 48 h. The embryo development to hatching blastocyst was significantly lower for the group with 10% rhLH versus that of the control group (65.6 vs. 79.5 for rhFSH+10% rhLH vs. control, respectively). However, no significant difference was found in development to hatching blastocyst for the control, rhFSH alone, and rhFSH+5% rhLH groups. The developmental potential of frozen-thawed embryos obtained from all groups was similar, with an 83.5% in vitro development rate until the expanded blastocyst stage. To detect anti-FSH antibodies, in Experiment 2, does were subject to four superovulation treatments. The hormone administration had a significant effect on immune response in the superovulation group after two treatments (0.14+/-0.074 and 0.15+/-0.076 vs. 0.46+/-0.078 and 0.50+/-0.078 optical density for the first, second, third, and forth cycles, respectively). Nevertheless, none of the treated does had an immune response in both the first and second treatments; on the contrary, a significant increase in the antibody levels was observed in these females at the moment of the third and fourth superovulation treatments. In conclusion, rhFSH superovulation treatments increase the reproductive potential of rabbit does. PMID- 19559475 TI - The Youth Psychopathic traits Inventory: psychometric properties and its relation to substance use and interpersonal style in a Dutch sample of non-referred adolescents. AB - This study presents evidence on the reliability and construct validity of the Dutch version of the Youth Psychopathic traits Inventory (YPI), a self-report instrument for psychopathic traits in adolescent boys and girls. In a sample of 776 Dutch non-referred adolescents, the YPI was found to have good internal consistency. Furthermore, exploratory Principal Components Analysis (PCA) indicated a three-factor structure for the YPI, including a Grandiose manipulative, Callous-unemotional, and Impulsive-irresponsible dimension. Good internal consistency was demonstrated for all dimensions in boys as well as girls, except for the Callous-unemotional dimension. Results further indicated strong correlations between psychopathic traits and dominant and hostile interpersonal style. The Grandiose-manipulative dimension predicted dominant interpersonal behavior in boys, but not in girls. In girls, a dominant interpersonal style was predicted by the Impulsive-irresponsible dimension. Psychopathic traits, mainly the Impulsive-irresponsible dimension, were related to a higher frequency of self-reported drug and alcohol use in boys and girls. PMID- 19559476 TI - Nifurtimox-eflornithine combination therapy for second-stage African Trypanosoma brucei gambiense trypanosomiasis: a multicentre, randomised, phase III, non inferiority trial. AB - BACKGROUND: Human African trypanosomiasis (HAT; sleeping sickness) caused by Trypanosoma brucei gambiense is a fatal disease. Current treatment options for patients with second-stage disease are toxic, ineffective, or impractical. We assessed the efficacy and safety of nifurtimox-eflornithine combination therapy (NECT) for second-stage disease compared with the standard eflornithine regimen. METHODS: A multicentre, randomised, open-label, active control, phase III, non inferiority trial was done at four HAT treatment centres in the Republic of the Congo and the Democratic Republic of the Congo. Patients aged 15 years or older with confirmed second-stage T b gambiense infection were randomly assigned by computer-generated randomisation sequence to receive intravenous eflornithine (400 mg/kg per day, every 6 h; n=144) for 14 days or intravenous eflornithine (400 mg/kg per day, every 12 h) for 7 days with oral nifurtimox (15 mg/kg per day, every 8 h) for 10 days (NECT; n=143). The primary endpoint was cure (defined as absence of trypanosomes in body fluids and a leucocyte count T; SNP2: EF_647729.1:g.405C>T; SNP3: EF_647729.1: g.433A>G and SNP4: EF_647729.1:g.588A>G), which showed significant frequency differences among 99 bovines of each species (n=198). Characterization of the isolated region revealed the presence of 6 putative haplotypes, 14 genotypes, which are formed by haplotypes, and numerous putative transcription factor binding sites. Only the thymine presence on SNPs 1 and 2, more common in B. indicus, was observed to alter some of the sites in this region. Despite this fact, analyses through real time PCR on bovines that present the most common homozygote genotypes of each species, which contrast for all the polymorphism, revealed no difference on the DARC gene transcription. Thus, in principle, it was concluded that the polymorphisms identified would not be useful as molecular markers in an improvement program for resistance to babesiosis. PMID- 19559489 TI - Adrenoreceptor-coupled signal-transduction mechanisms mediating lymphocyte apoptosis induced by endogenous catecholamines. AB - Our previous work has shown that lymphocytes synthesize catecholamines (CAs) and the endogenous CAs accelerate apoptosis of concanavalin A (Con A)-activated lymphocyte. Here, we explored the involvement of adrenoreceptors (ARs) and signal molecules coupled to the ARs in the endogenous CA-mediated modulation of lymphocyte apoptosis. Pargyline, an inhibitor of CA degradation, up-regulated the expression of cleaved caspase-3 protein and increased the number of apoptotic cells. Antagonists of alpha(1)-ARs and beta(2)-ARs, not antagonists of alpha(2) ARs or beta(1)-ARs, blocked these effects of pargyline. The facilitating effects of pargyline on lymphocyte apoptosis were mimicked by activators of adenylate cyclase and PKC, but reversed by inhibitors of PKA, PLC and PKC. Pargyline stimulated CREB activation and Smac/DIABLO expression were prevented by the inhibitors of PKA, PLC and PKC. These results imply that endogenous CA-induced lymphocyte apoptosis is mediated by cAMP-PKA- and PLC-PKC-linked CREB-Smac/DIABLO pathways coupled with alpha(1)-ARs and beta(2)-ARs. PMID- 19559491 TI - Early abciximab administration before primary percutaneous coronary intervention improves clinical outcome in elderly patients transferred with ST-elevation myocardial infarction: data from the EUROTRANSFER registry. AB - BACKGROUND: Limited data are available concerning benefits and risks of early abciximab (EA) administration before primary percutaneous coronary intervention (PPCI) in elderly ST-segment elevation myocardial infarction (STEMI) patients. The objective of the study was to assess the impact of EA before PPCI in elderly (>or=65 years) patients. METHODS AND RESULTS: We identified 545 patients <65 years (354 with EA administration (>30 min before PPCI), 191 late abciximab (LA)), and 541 patients >or=65 years of age (373 EA, 168 LA) in the EUROTRANSFER Registry database. Elderly patients were more likely to have comorbidities, angiographic PCI complications, and bleeding events. EA promotes infarct-related artery patency before PPCI and improves myocardial reperfusion after PPCI in both age groups, but the risk of 30-day death (EA vs. LA: <65 years, 2.0% vs. 1.6%; p=0.999; >or=65 years, 5.9% vs. 14.3%; p=0.001) and 30-day death+reinfarction (EA vs. LA: <65 years, 2.5% vs. 2.1%; p=0.999; >or=65 years, 7.5% vs. 17.3%; p=0.001) was reduced in elderly patients only. There was no difference in bleedings, especially major bleedings requiring transfusion (EA vs. LA: patients <65 years, 2.3% vs. 0%, p=0.055; >or=65 years, 2.4% vs. 3%; p=0.448) between groups. CONCLUSIONS: Patients >or=65 years of age have a substantially increased risk of angiographic PCI complications, death and bleeding events compared with their younger counterparts. Strategy of EA before PPCI improves reperfusion parameters and clinical outcome in elderly patients and is not associated with elevated risk of major bleeding. PMID- 19559492 TI - Co-morbidity index predicts for mortality after stereotactic body radiotherapy for medically inoperable early-stage non-small cell lung cancer. AB - PURPOSE: To determine the prognostic role of co-morbidity in medically inoperable early-stage non-small cell lung cancer (NSCLC) treated with stereotactic body radiotherapy (SBRT). METHODS AND MATERIALS: Between 2000 and 2007, 88 consecutive early-stage medically inoperable NSCLC patients were treated by linac-based SBRT. The dose was either 45 Gy or 67.5 Gy in three fractions prescribed to the isocenter. Baseline co-morbidities were retrospectively retrieved by consultation of a formal electronic registry of diagnoses as well as patients' charts. The age adjusted Charlson Co-morbidity Index (CCI) was scored for each patient and subjected to univariate and multivariate analysis. RESULTS: With a median follow up of 44 months, the actuarial local control rate at 4 years was 89% while the median overall survival was 22 months. The median age-adjusted CCI score was 5. The age-adjusted CCI was a significant predictor of overall survival on both univariate (p=0.002) and multivariate analysis (p=0.011). Patients with an age adjusted CCI score of 3 or less had a median survival of 41 months versus only 11 months for those scoring 6 or more. CONCLUSION: The number and seriousness of co morbidities predict overall survival in medically inoperable early-stage NSCLC treated with SBRT. Because the determination of medical operability is frequently based on both objective measures and subjective clinical judgment, it is recommended that co-morbidity be formally indexed in all studies examining the outcomes of SBRT. PMID- 19559490 TI - Synergy of TRIF-dependent TLR3 and MyD88-dependent TLR7 in up-regulating expression of mouse FPR2, a promiscuous G-protein-coupled receptor, in microglial cells. AB - Human G-protein-coupled formyl peptide receptor-like 1 and its mouse homologue formyl peptide receptor 2 mediate the chemotactic activity of a variety of pathogen and host-derived peptides, including amyloid beta(42), a key causative factor in Alzheimer's disease (AD).Here, we found that polyinosine-polycytidylic acid (Poly(I:C)), which is a specific TLR3 ligand, and Imiquimod (R837), which is a specific TLR7 ligand, when used alone, each increased MAPK-dependent functional mFPR2 expression in microglial cells, and the combination of Poly(I:C) and R837 exhibited additive effect by enhancing the level of IkappaB-alpha phosphorylation. Our results indicated that RNA virus infection may actively participate in the pathogenic processes of brain inflammation and neurodegenerative diseases by TLR3- and TLR7-mediated TRIF-dependent and MyD88 dependent signaling pathways. PMID- 19559493 TI - Contribution of the CesR-regulated genes llmg0169 and llmg2164-2163 to Lactococcus lactis fitness. AB - Lactococcus lactis is one of the main components of the starter cultures used in cheese manufacture. As starter, L. lactis must tolerate harsh conditions encountered either during their production in bulk quantities or during dairy products processing. To face these hostile conditions, bacteria monitor the environment and respond by modifying gene expression appropriately. Previous transcriptomic studies showed that the two component system CesSR is the main pathway that triggers the cell envelope stress response in L. lactis treated with lactococcin 972 (Lcn972), a cell wall synthesis inhibiting bacteriocin. Among the CesR-regulated genes, llmg0169 and the operon llmg2164-2163, encoding proteins of unknown function, are among the highest up-regulated genes after activation of CesSR. In this study, we have assessed the contribution of these genes to the survival of L. lactis to different technologically-relevant stresses. Overexpressing and knock-out mutants of the genes were generated and their viability to low pH, heat, freeze-drying, presence of NaCl, cell wall antimicrobials and lytic phages attack was compared to the wild type strain. The genes llmg0169 and llmg2164-2163 contributed differently to L. lactis fitness. L. lactis Deltallmg0169 was very sensitive to heat treatment while L. lactis Deltallmg2164 was more sensitive to NaCl. Absence of both genes also compromised viability at low pH. On the contrary, higher expression levels of llmg0169 and llmg2164-2163, up to 26- and 14-fold increase determined by qRT-PCR, respectively, did not enhance L. lactis survival in any of the above stressful conditions (heat, pH and NaCl) or after freeze-drying. All the mutants displayed a similar phage susceptibility profile. Overexpression of llmg2164-2163 seemed to specifically protect L. lactis against the bacteriocin Lcn972 but not against other cell wall active antimicrobials. Based on our phenotypic analysis, the investigated genes are required to mount a proper response to guarantee survival of L. lactis under technologically-relevant stresses and their functionality could be a useful marker to select robust dairy starters. PMID- 19559494 TI - Non-alcoholic fatty liver disease (NAFLD) and mortality. PMID- 19559495 TI - Where are healthcare providers? Exploring relationships between context and human resources for health Madhya Pradesh province, India. AB - OBJECTIVE: In India, heterogeneous healthcare providers in the public and dominant private sectors serve a diverse population, including those from vulnerable groups, the scheduled castes and tribes. We explored relationships between the distribution of different categories of healthcare providers (public and private); and contextual socioeconomic and demographic variables. Access to healthcare providers for scheduled castes and tribes was specifically studied. METHOD: Set in Madhya Pradesh province (60.4 million), India. Dependent variables included district-wise densities of physicians and paramedics (public and private separately); and unqualified providers (private). Contextual variables included infrastructure, urbanization, economy, female literacy and proportion of scheduled castes and tribes. RESULTS: Urbanization was strongly correlated with private physician density; and negatively with paramedical density (public and private). Private paramedical density variation was partially explained by economy. Public physician and paramedical density were positively correlated to district proportions of scheduled tribes. All provider densities (public and private) were negatively related to proportions of scheduled castes. CONCLUSIONS: Overall density of qualified providers was low. Qualified physicians tended to be more densely situated in the relatively more urban districts. Access to healthcare providers for scheduled castes and tribes is different. More targeted approaches are necessary for improving access for scheduled castes. PMID- 19559496 TI - Limitations of dual time point PET in the assessment of lung nodules with low FDG avidity. AB - FDG PET has long shown efficacy in the evaluation of indeterminate pulmonary nodules. More recently, the use of dual time point imaging has been looked at as a means for improving sensitivity and accuracy. While initial reports were very promising, more recent results looking specifically at pulmonary lesions with low levels of FDG avidity demonstrated limitations. These lesions (initial maximum standard uptake value of less than 2.5) are of particular interest due to the fact that well-differentiated adenocarcinomas, broncheoaveolar carcinoma and carcinoid may have low FDG avidity on standard PET imaging, leading to false negative exams. Our study retrospectively reviewed the accuracy of dual time point (DTP) FDG PET imaging to determine if it aided in the identification of malignant pulmonary nodules when initial time point imaging showed a maximum SUV of less than 2.5. 113 patients had undergone a total of 130 DTP PET/CT with 152 lesions assessed. 67 lesions were subsequently definitively diagnosed as benign or malignant based upon biopsy or imaging follow-up. Utilizing a maximum SUV increase of 10%, which optimizes our sensitivity and specificity; our results demonstrate a sensitivity of 63% and a specificity of 59%, similar to other investigators evaluating lesions with low FDG avidity. Increasing or decreasing this threshold did not improve our results, nor did the addition of lesions with maximum SUV's of 2.5 or greater on initial imaging. Specifically in nodules with low FDG avidity (max SUV<2.5), the sensitivity was 61%, specificity 58%, and accuracy was 60%. Our findings suggest that DTP FDG PET may not be of benefit in the assessment of pulmonary nodules with maximum SUV of less than 2.5 on initial imaging. PMID- 19559497 TI - LAT1 expression in non-small-cell lung carcinomas: analyses by semiquantitative reverse transcription-PCR (237 cases) and immunohistochemistry (295 cases). AB - OBJECTIVE: System l-amino acid transport mediates the uptake of aromatic neutral amino acids and nutritionally essential amino acids from extracellular fluids. Little is known about the role of l-amino acid transporter 1 (LAT1), a member of the system l-amino acid transporter family, in non-small-cell lung carcinomas (NSCLCs). PATIENTS AND METHODS: We examined (i) LAT1 mRNA levels in 40 normal lung tissues (NLTs) and 237 NSCLCs using semiquantitative RT-PCR, (ii) LAT1 protein expression in 295 NSCLCs using immunohistochemistry, and (iii) whether LAT1 mRNA and protein expressions were related to clinicopathologic findings and outcome. RESULTS: The LAT1 mRNA level was significantly higher in all NSCLCs (6.81+/-1.13) than in NLT (1.00+/-0.18). The LAT1 mRNA level showed no association with clinicopathologic findings or outcome. LAT1 protein was detected with a diffuse or granular appearance within the cytoplasm and/or on the plasma membrane of tumor cells. When tumors were graded as positive if staining indicating a plasma membrane expression of LAT1 protein made up more than 10% of the tumor, the frequency of this membrane expression was found to be associated with tumor histology, differentiation grade, pathologic stage, T classification, pleural invasion, lymph-vessel invasion, and overall survival rate. CONCLUSION: Detection of a plasma membrane expression of LAT1 protein would appear to be of value in informing the prognosis in NSCLC cases. PMID- 19559498 TI - Comparison of squamous cell carcinoma and adenocarcinoma of the lung by metabolomic analysis of tissue-serum pairs. AB - The prospect of establishing serum metabolomic profiles offers great clinical significance for its potential to detect human lung cancers at clinically asymptomatic stages. Patients with suspicious serum metabolomic profiles may undergo advanced radiological tests that are too expensive to be employed as screening tools for the mass population. As the first step to establishing such profiles, this study investigates correlations between tissue and serum metabolomic profiles for squamous cell carcinoma (SCC) and adenocarcinoma (AC) in the lungs of humans. Tissue and serum paired samples from 14 patients (five SCCs and nine ACs), and seven serum samples from healthy controls were analyzed with high-resolution magic angle spinning proton magnetic resonance spectroscopy (HRMAS (1)HMRS). Tissue samples were subjected to quantitative histological pathology analyses after MRS. Based on pathology results, tissue metabolomic profiles for the evaluated cancer types were established using principal component and canonical analyses on measurable metabolites. The parameters used to construct tissue cancer profiles were then tested with serum spectroscopic results for their ability to differentiate between cancer types and identify cancer from controls. In addition, serum spectroscopic results were also analyzed independent of tissue data. Our results strongly indicate the potential of serum MR spectroscopy to achieve the task of differentiating between the tested human lung cancer types and from controls. PMID- 19559499 TI - Multi-stage optimal design for groundwater remediation: a hybrid bi-level programming approach. AB - This paper presents the development of a hybrid bi-level programming approach for supporting multi-stage groundwater remediation design. To investigate remediation performances, a subsurface model was employed to simulate contaminant transport. A mixed-integer nonlinear optimization model was formulated in order to evaluate different remediation strategies. Multivariate relationships based on a filtered stepwise clustering analysis were developed to facilitate the incorporation of a simulation model within a nonlinear optimization framework. By using the developed statistical relationships, predictions needed for calculating the objective function value can be quickly obtained during the search process. The main advantage of the developed approach is that the remediation strategy can be adjusted from stage to stage, which makes the optimization more realistic. The proposed approach was examined through its application to a real-world aquifer remediation case in western Canada. The optimization results based on this application can help the decision makers to comprehensively evaluate remediation performance. PMID- 19559500 TI - Lidocaine inhibits epithelial chemokine secretion via inhibition of nuclear factor kappa B activation. AB - AIM: To study the antiinflammatory effect of lidocaine in intestinal epithelial cells. METHODS: HT-29 and T-84 cells were grown in culture with and without TNF alpha, lidocaine, aconitine and veratridine. The secretion of IL-8 and IP-10 was measured by ELISA. A cDNA microarray was used to assess gene expression. Real time PCR was used to confirm the results. Western blots and a modified electromobility shift assay (EMSA) were used to assess NFkappaB activation. RESULTS: Lidocaine inhibited spontaneous and TNF-alpha induced secretion of IL-8 and IP-10. The combination of veratridine or aconitine, voltage-gated sodium channels (VGSC) agonists that open VGSCs, with lidocaine did not alter the effect of lidocaine on cytokine secretion. Gene array analysis revealed that IkappaB transcription was induced by TNF-alpha and inhibited by lidocaine. IkappaB real time PCR confirmed this observation. A Western blot analysis demonstrated that the degradation of IkappaB following TNF-alpha treatment was markedly inhibited by lidocaine. Lidocaine treatment resulted in decreased generation of phosphorylated IkappaB. A modified EMSA was complementary and demonstrated marked inhibition of NFkappaB nuclear binding. CONCLUSION: Lidocaine inhibits IL-8 and IP-10 secretion from intestinal cells. This effect is mediated by inhibition of NFkappaB activation via decreased IkappaB phosphorylation and is not mediated by lidocaine's effect on VGSC. PMID- 19559501 TI - Differential glycosaminoglycan expression and hyaluronan homeostasis in juvenile hyaline fibromatosis. AB - BACKGROUND: Juvenile hyaline fibromatosis (JHF) is a rare autosomal recessive disease characterized histologically by deposition of hyaline material and clinically by multiple skin lesions. Clarification of the molecular and structural changes involved in JHF skin lesions may unravel targets for pharmacotherapy. OBJECTIVE: We sought to investigate the expression of glycosaminoglycans and their metabolizing enzymes in lesional as compared with lesion-free skin tissue specimens in JHF. METHODS: Glycosaminoglycans were isolated, purified, and fractionated by electrophoresis on cellulose acetate membranes and agarose gels. Hyaluronic acid (HA) was quantitated by enzyme-linked immunosorbent assay and the expression of HA metabolizing enzymes was investigated using reverse transcriptase-polypeptide chain reaction. RESULTS: JHF lesions exhibited significantly less HA and elevated amounts of dermatan sulfate and chondroitin sulfate, whereas gene expression of HA synthase-1 and HA synthase 3 was significantly down-regulated, as compared with lesion-free skin tissue specimens. LIMITATIONS: Because JHF is a rare disease, a limitation to our study was that we collected skin tissue specimens from only one patient. CONCLUSION: The significant alterations of HA homeostasis in JHF lesions provide further understanding of JHF pathogenesis and may offer a target for pharmacologic intervention to treat the skin lesions associated with JHF. PMID- 19559502 TI - Risk factors for peripheral intravenous catheter infection in hospitalized patients: a prospective study of 3165 patients. AB - We conducted a prospective study of 6538 polyurethane peripheral intravenous (IV) catheters in 3165 hospitalized adult patients using semiquantitative culture techniques. We found that extending the scheduled catheter replacement interval from 48 to 72 hours to 72 to 96 hours was not a risk factor for local catheter infection, but that catheter insertion by personnel other than IV therapists and the use of continuous infusion to maintain catheter patency were 2 independent risk factors for infection. PMID- 19559503 TI - Successful prevention of ventilator-associated pneumonia in an intensive care setting. AB - BACKGROUND: Ventilator-associated pneumonia (VAP) is one of the most common health care-associated infections (HAIs) in critical care settings. OBJECTIVE: Our objective was to examine the effect of a series of interventions, implemented in 3 different periods to reduce the incidence of VAP in an intensive care unit (ICU). METHODS: A quasiexperimental study was conducted in a medical-surgical ICU. Multiple interventions to optimize VAP prevention were performed during different phases. From March 2001 to December 2002 (phase 1: P1), some Centers for Disease Control and Prevention (CDC) evidence-based practices were implemented. From January 2003 to December 2006 (P2), we intervened in these processes at the same time that performance monitoring was occurring at the bedside, and, from January 2007 to September 2008 (P3), we continued P2 interventions and implemented the Institute for Healthcare Improvement's ventilator bundle plus oral decontamination with chlorhexidine and continuous aspiration of subglottic secretions. RESULTS: The incidence density of VAP in the ICU per 1000 patient-days was 16.4 in phase 1, 15.0 in phase 2, and 10.4 in phase 3, P=.05. Getting to zero VAP was possible only in P3 when compliance with all interventions exceeded 95%. CONCLUSION: These results suggest that reducing VAP rates to zero is a complex process that involves multiple performance measures and interventions. PMID- 19559504 TI - Use of atmospheric non-thermal plasma as a disinfectant for objects contaminated with methicillin-resistant Staphylococcus aureus. AB - BACKGROUND: Health care-associated infections because of methicillin-resistant strains of Staphylococcus aureus (MRSA) are increasing worldwide despite current infection control measures. Novel methods for disinfection of MRSA would be useful. METHODS: We tested the effectiveness of atmospheric, non-thermal plasma discharge at killing S aureus, including USA300 strains, and at disinfecting experimentally contaminated hospital pagers. RESULTS: Exposure of S aureus to plasma at different concentrations and for varying lengths of time resulted in up to a 4- to 5-log(10) kill on tryptic soy agar plates within 10 minutes and was not toxic to epithelial cells. USA300 strains of MRSA were more resistant to plasma-based killing than other tested strains. Disinfection of hospital pagers experimentally coated with clinically relevant amounts of MRSA could be achieved in as little as 30 seconds. CONCLUSION: Generation of plasma is a promising method for disinfection of objects or surfaces that warrants further study in hospital settings. The USA300 strains of S aureus may be more resistant to disinfection than other strains. PMID- 19559506 TI - [Students, graduates and family medicine: elements for an imperfect relationship]. PMID- 19559508 TI - Student centred teaching methods in a Chinese setting. AB - This paper offers a discussion about using Western, student centred teaching methods with Chinese student nurses. There is increasing interest from Chinese nurse educators in student centred learning and an increase in partnerships between Chinese and Western universities. This paper suggests that the assumption that Western teaching methods are superior is now questioned and transferring Western style teaching to China requires a high degree of cultural sensitivity. PMID- 19559505 TI - Effects of age, genes, and pulse pressure on executive functions in healthy adults. AB - Executive functions (EF) evidence significant age-related declines, but the mechanisms underpinning those changes are unclear. In this study, we focus on two potential sources of variation: a physiological indicator of vascular health, and genetic variants related to vascular functions. In a sample of healthy adults (n=158, ages 18-81), we examine the effects of age, pulse pressure, and two polymorphisms (comt val158met and ace insertion/deletion) on working memory and cognitive flexibility. Results indicate that in addition to often-replicated age differences, the alleles of two polymorphisms that promote vasoconstriction (comt val and ace D) and reduced availability of dopamine in neocortical synapses (comt val), negatively impact virtually all aspects of EF tasks that involve working memory. In some cases, suppression of cognitive performance is limited to men or necessitates a combination of both risk-associated alleles. After accounting for genetic and age-related variation, pulse pressure had no additional effect on EF. These findings suggest that in healthy adults, the effects of genetic risk factors significantly modulate the course of cognitive aging. PMID- 19559509 TI - Uranium isotopes in groundwater from the continental intercalaire aquifer in Algerian Tunisian Sahara (Northern Africa). AB - The disequilibrium between (234)U and (238)U is commonly used as a tracer of groundwater flow. This paper aims to identify uranium contents and uranium isotopic disequilibria variation in groundwater sampled from deep Continental Intercalaire aquifer (southern Algeria and Tunisia). Large variations in both U contents (0.006-3.39ppb) and (234)U/(238)U activity ratios (0.4-15.38) are observed. We conduct a first assessment in order to verify whether the results of our investigation support and complete previous hydrogeological and isotopic studies. The dissolved U content and (234)U/(238)U activity ratio data were plotted on a two-dimensional diagram that was successfully utilized on sharing the CI aquifer into different compartments submitted to different oxidising/reducing conditions and leads also to distinguished two preferential flow paths in the Nefzaoua/Chott Fejej discharge area. Uranium isotopes disequilibrium indicate that ranium chemistry is mainly controlled by water-rock interaction enhanced by long residence time recognised for this aquifer. PMID- 19559507 TI - Metals in anticancer therapy: copper(II) complexes as inhibitors of the 20S proteasome. AB - Selective 20S proteasomal inhibition and apoptosis induction were observed when several lines of cancer cells were treated with a series of copper complexes described as [Cu(L(I))Cl] (1), [Cu(L(I))OAc] (2), and [Cu(HL(I))(L(I))]OAc (3), where HL(I) is the ligand 2,4-diiodo-6-((pyridine-2-ylmethylamino)methyl)phenol. These complexes were synthesized, characterized by means of ESI spectrometry, infrared, UV-visible and EPR spectroscopies, and X-ray diffraction when possible. After full characterization species 1-3 were evaluated for their ability to function as proteasome inhibitors and apoptosis inducers in C4-2B and PC-3 human prostate cancer cells and MCF-10A normal cells. With distinct stoichiometries and protonation states, this series suggests the assignment of species [CuL(I)](+) as the minimal pharmacophore needed for proteasomal chymotryspin-like activity inhibition and permits some initial inference of mechanistic information. PMID- 19559510 TI - Sediment budgets and source determinations using fallout Cesium-137 in a semiarid rangeland watershed, Arizona, USA. AB - Analysis of soil redistribution and sediment sources in semiarid and arid watersheds provides information for implementing management practices to improve rangeland conditions and reduce sediment loads to streams. The purpose of this research was to develop sediment budgets and identify potential sediment sources using (137)Cs and other soil properties in a series of small semiarid subwatersheds on the USDA ARS Walnut Gulch Experimental Watershed near Tombstone, Arizona, USA. Soils were sampled in a grid pattern on two small subwatersheds and along transects associated with soils and geomorphology on six larger subwatersheds. Soil samples were analyzed for (137)Cs and selected physical and chemical properties (i.e., bulk density, rocks, particle size, soil organic carbon). Suspended sediment samples collected at measuring flume sites on the Walnut Gulch Experimental Watershed were also analyzed for these properties. Soil redistribution measured using (137)Cs inventories for a small shrub-dominated subwatershed and a small grass-dominated subwatershed found eroding areas in these subwatersheds were losing -5.6 and -3.2tha(-1)yr(-1), respectively; however, a sediment budget for each of these subwatersheds, including depositional areas, found net soil loss to be -4.3tha(-1)yr(-1) from the shrub dominated subwatershed and -0.1tha(-1)yr(-1) from the grass-dominated subwatershed. Generally, the suspended sediment collected at the flumes of the six other subwatersheds was enriched in silt and clay. Using a mixing model to determine sediment source indicated that shrub-dominated subwatersheds were contributing most of the suspended sediment that was measured at the outlet flume of the Walnut Gulch Experimental Watershed. The two methodologies (sediment budgets and sediment source analyses) indicate that shrub-dominated systems provide more suspended sediment to the stream systems. The sediment budget studies also suggest that sediment yields measured at the outlet of a watershed may be a poor indicator of actual soil redistribution rates within these semiarid watersheds. Management of these semiarid rangelands must consider techniques that will protect grass-dominated areas from shrub invasion to improve rangeland conditions. PMID- 19559511 TI - Prognostic significance of erythropoietin and erythropoietin receptor in tongue squamous cell carcinoma. AB - Despite its primary hematopoietic function, erythropoietin (Epo) is a pleiotropic cytokine that exerts various biological functions in many different non hematopoietic cells and cancers, and its stimulatory effects are mediated through activation of its receptor (EpoR). Recent studies have shown that Epo and EpoR may be involved in carcinogenesis, angiogenesis, and invasion. We have investigated the expression of Epo and EpoR in a series of 65 resected squamous cell carcinomas (SCC) of the tongue using immunohistochemical staining. The proportion of Epo and EpoR changes in them was greater than those in normal squamous epithelium (P<0.05). Epo expression was associated with age, density of microvessels, and the stage of the tumour (P<0.05). EpoR expression was associated with microvascular density alone (P<0.05). After adjusting for other clinicopathological factors, Epo and EpoR expression remained independent adverse prognosticators for postoperative survival (P<0.05). Our findings support the hypothesis that the Epo and EpoR systems influence the prognosis of carcinogenesis, angiogenesis, and malignant progression of SCC of the tongue, and confirm that Epo and EpoR are independent prognostic markers. PMID- 19559512 TI - Counseling and choosing between infant-feeding options: overall limits and local interpretations by health care providers and women living with HIV in resource poor countries (Burkina Faso, Cambodia, Cameroon). AB - As part of prevention of HIV mother-to-child transmission (PMTCT) strategies, HIV positive women are asked to choose between two options regarding infant feeding: replacement feeding or exclusive breastfeeding with early weaning. Health services must offer women counseling, guidance, and support to enable them to make an informed choice. This article aims to shed light on the content of counseling and its adaptation to local situations, including women's perceptions, in three resource-poor countries with different infant feeding patterns (Burkina Faso, Cambodia, and Cameroon). The qualitative study included observations in health facilities and interviews with HIV-positive mothers and health workers. The results show that counseling practices vary, including prescriptive counseling proposing only one option to all women, an option based on the mother's economic level assessed by health care providers, and the choice between options. While health workers consider economic aspects first, women mostly consider social aspects related to the risk of being stigmatized as a "bad mother'" or as HIV-positive. Overall trends identify some limits to counseling effectiveness when considering women's perceptions and needs, such as: the content of information provided by health care providers, duration and timing of counseling, discrepancies between information provided during counseling and from the health system, and ranking of preventive options by health workers. Women's agency for feeding choices is related to local practices and local authorities' abilities to provide more or less comprehensive counseling based on the organization of the health and aid system. Local practices also depend on institutions' interpretations of international recommendations based on public health considerations regarding standard of care and women's and the health system's respective responsibilities. Beyond structural constraints that hinder the adoption of preventive infant feeding patterns, addressing these issues may help improve counseling practices. PMID- 19559513 TI - "You don't need a prescription to go gluten-free": the scientific self-diagnosis of celiac disease. AB - We explore the social process of celiac disease diagnosis using fieldwork in the United States with two celiac support groups, interviews, and a virtual ethnography of an online discussion board. Distinguishing between medical diagnosis, self-diagnosis, and scientific self-diagnosis, we examine patients' varied paths to diagnosis and their attempts to legitimize symptoms as celiac disease. Web-based direct-access testing (DAT) permits patients to bypass physician requisition for testing in their diagnostic quest. While such laboratories do not diagnose disease per se, they provide the consumer with the scientific information necessary to self-diagnose. This scientific self-diagnosis grants individuals greater legitimacy for their claims of an illness identity than self-diagnosis alone, but less legitimacy than medical diagnosis. We examine the implications of scientific self-diagnosis for the social construction of diagnosis and professional and lay ways of knowing. PMID- 19559514 TI - Responses on a lateralized lexical decision task relate to both reading times and comprehension. AB - Research over the last few years has shown that the dominance of the left hemisphere in language processing is less complete than previously thought [Beeman, M. (1993). Semantic processing in the right hemisphere may contribute to drawing inferences from discourse. Brain and Language, 44, 80-120; Faust, M., & Chiarello, C. (1998). Sentence context and lexical ambiguity resolution by the two hemispheres. Neuropsychologia, 36(9), 827-835; Weems, S. A., & Zaidel, E. (2004). The relationship between reading ability and lateralized lexical decision. Brain and Cognition, 55(3), 507-515]. Engaging the right brain in language processing is required for processing speaker/writer intention, particularly in those subtle interpretive processes that help in deciphering humor, irony, and emotional inference. In two experiments employing a divided field or lateralized lexical decision task (LLDT), accuracy and reaction times (RTs) were related to reading times and comprehension on sentence reading. Differences seen in RTs and error rates by visual fields were found to relate to performance. Smaller differences in performance between fields tended to be related to better performance on the LLDT in both experiments and, in Experiment 1, to reading measures. Readers who can exploit both hemispheres for language processing equally appear to be at an advantage in lexical access and possibly also in reading performance. PMID- 19559515 TI - A qualitative study on patient perspectives of how conventional and implant supported dentures affect eating. AB - OBJECTIVES: The objectives of this study were to obtain in-depth qualitative data from patients on the effect of edentulousness and prosthetic rehabilitation upon issues surrounding eating (emotional, social and functional). METHODS: 33 patients who had received replacement conventional dentures (CDs) and 33 patients who had received implant-supported mandibular overdentures (ISODs) were interviewed by a researcher outside the clinical team. Interviews focused upon their experience of edentulousness, its management and its impact on their lives. Interviews were transcribed and the qualitative data analysed using NVivo [QSR NVivo v.2.2. Melbourne: QSR International; 2002]. RESULTS: Both groups of patients told similar stories about the impact of edentulousness upon eating. A few patients who had received replacement CDs reported significant improvement, but marginal improvements were more common. Most patients who had received ISODs reported a significant positive impact upon eating with very few negative effects reported. The findings suggest that the functional limitations of dentures often impose social restrictions on edentulous patients. Optimising conventional dentures may help, but patients' eating-related quality of life is most likely to be enhanced through the functional improvement and increased social confidence that ISODs bring. CONCLUSIONS: In conclusion, issues surrounding eating are a major concern for edentulous patients. The main impacts of edentulousness are limitation of social participation and food choice. The functional improvement and increased social confidence that ISODs can provide, has a beneficial impact on this aspect of patients of quality of life. PMID- 19559516 TI - Comment on: Management of pPROM before 34 weeks' gestation. PMID- 19559517 TI - Cox-2 expression in ovarian malignancies: a review of the clinical aspects. AB - COX-2 is an inducible enzyme expressed only in response to stimuli such as mitogens, cytokines, growth factors or hormones, and is pro-inflammatory. It plays an important role in tumorigenesis. The purpose of the present report is to review the clinical aspects of COX-2 expression in ovarian malignancies. A PubMed (http://www.pubmed.gov/) search of investigations published from July 2001 until August 2008 and containing the term COX-2 in combination with ovarian malignancies was conducted. The clinical aspects of the relevant investigations were reviewed. COX-2 is expressed in ovarian tumors of low malignant potential (LMP), in epithelial ovarian carcinoma (EOC) and primary peritoneal carcinoma (PPC) and apparently plays a role in their carcinogenesis. Its expression seems to be correlated with VEGF that serves as a predictor of poor prognosis in some non-gynecologic malignancies. COX-2 expression is higher in EOC than in LMPs. The results with regard to the association between COX-2 expression and prognostic factors, response to treatment and outcome in ovarian malignancies are inconsistent. Clinical studies dealing with the effect of COX-2 inhibitors on outcome are scarce. The use of COX-2 expression in gynecological malignancies in clinical practice remains to be elucidated. PMID- 19559518 TI - The extent of lymphadenectomy seems to be associated with better survival in patients with nonmetastatic upper-tract urothelial carcinoma: how many lymph nodes should be removed? AB - BACKGROUND: The role and extent of lymphadenectomy in patients with upper-tract urothelial carcinoma (UTUC) is debated. OBJECTIVE: To establish whether the number of lymph nodes (LNs) removed might be associated with better cause specific survival in patients with UTUC. DESIGN, SETTING, AND PARTICIPANTS: The study included 552 consecutive patients who underwent radical nephroureterectomy (RNU) and lymphadenectomy between 1992 and 2006. INTERVENTION: Patients were treated with RNU and lymphadenectomy. MEASUREMENTS: Univariable and multivariable Cox proportional hazards regression models addressed the association between the number of LNs removed and cause-specific mortality (CSM). The number of LNs removed was coded as a cubic spline to allow for nonlinear effects. Finally, the most informative cut-off for the number of removed LNs was identified. RESULTS AND LIMITATIONS: In the entire population, the number of LNs removed was not associated with CSM in univariable (hazard ratio [HR]: 0.99; p=0.16) or in multivariable (HR: 0.97; p=0.12) analyses. In contrast, in the subgroup of pN0 patients (n=412), the number of LNs removed achieved the independent predictor status of CSM (HR: 0.93; p=0.02). Eight LNs removed was the most informative cut off in predicting CSM (HR: 0.42; p=0.004). The inclusion of the variable defining dichotomously the number of removed LNs (< 8 vs > or = 8) in the base model (age, Eastern Cooperative Oncology Group performance status, pathologic stage, grade, architecture, and lymphovascular invasion) significantly increased the accuracy in predicting CSM (+1.7%; p<0.001). CONCLUSIONS: The extension of the lymphadenectomy in pN0 UTUC patients seems to be associated with CSM. Longer survival was observed in patients in whom at least eight LNs had been removed. PMID- 19559519 TI - Editorial comment on: The extent of lymphadenectomy seems to be associated with better survival in patients with nonmetastatic upper-tract urothelial carcinoma: how many lymph nodes should be removed? PMID- 19559520 TI - Reduced life-space of non-professional caregivers to Parkinson's disease patients with increased disease duration. AB - OBJECTIVE: To evaluate the impact of Parkinson's disease (PD) on unpaid, non professional caregiver (CG) social relationships and related issues, as well as home-help services, over the time-course of the disease. MATERIALS AND METHODS: Cross-sectional telephone survey using a structured questionnaire where CGs were divided into three groups with regard to care-giving/PD duration: 0-4, 5-10 and >11 years. RESULTS: 404/451 CGs responded. Their relationships to friends, the patient, parents, other relatives, workmates and own children were impaired in 30%, 26%, 20%, 20%, 15% and 7% of the cases, respectively, with a significant increase in impaired parent and friends relations with the longest care-giving duration. More than half with the longest care-giving duration also experienced limitations in social activities, e.g. seeing friends and carrying out leisure activities. Of all CGs 11% used professional home-service help, with a mean of 8h assistance/week, however with no increased effect on possibilities to social activities. Of the 30% of CGs below the age of 65, one-third was on early retirement or sick leave and 40% of the remaining two-thirds worked part-time due to the patient-care commitment. Caregivers still working reported no more possibilities to social activities. CONCLUSIONS: Confinement caused by looking after PD patients may disorganize the life of the entire family and have adverse social consequences for unpaid CGs. This could disrupt the established balance within the family, limiting and narrowing the CGs' life-space. The narrowed life space of these CGs must be considered and acted upon when offering and planning healthcare. PMID- 19559521 TI - Treatment of limb-shaking TIAs with external carotid artery stenting. AB - Limb-shaking transient ischemic attacks (TIAs) occur when perfusion is reduced to the cortical territory associated with the transient movements. We present a case in which a patient with preexisting left internal carotid artery (ICA) occlusion developed limb-shaking TIAs secondary to severe stenosis of her left external carotid artery (ECA). After angioplasty and stenting of her left ECA, her symptoms resolved. PMID- 19559522 TI - Preparation of amine group-containing chelating fiber for thorough removal of mercury ions. AB - An aminated chelating fiber (AF) with high adsorption capacity for mercury ions was prepared by grafting copolymerization of acrylonitrile onto polypropylene fiber, followed by aminating with chelating molecule diethylenetriamine. Effects of reaction conditions such as temperature, reaction time, bath ratio and dosage of catalyst on the grafting yield were studied. Chemical structure, tensile strength and thermal stability of AF were characterized. The adsorption performances for mercury were evaluated by batch adsorption experiments and kinetic experiments. The results show that AF is effective for the removal of mercury over a wide range of pH. The chelating fiber also shows much higher adsorption capacities for mercury, the equilibrium adsorption amount could be as high as 657.9 mg/g for mercury. The high adsorption capacity of Hg(2+) on AF is resulted from the strong chelating interaction between amine groups and mercury ions. Two amine groups coordinate with one mercury ion could be speculated from the adsorption capacity and amine group content on AF. The kinetic adsorption results indicate that the adsorption rates of AF for mercury are very rapid. Furthermore, the residual concentration was less than 1 microg/L with feed concentration of mercury below 1mg/L, which can meet the criterion of drinking water, which indicates that the chelating fiber prepared in this study could be applied to low-level Hg contaminated drinking water purification. PMID- 19559523 TI - Electrochemical reduction of nitrate and nitrite in simulated liquid nuclear wastes. AB - The electrochemical reduction of nitrate and nitrite in simulated low-level nuclear wastes containing 1.8 M NaNO(3)+0.55 M NaNO(2)+1.16 M NaOH was studied under galvanostatic polarization on tin and bismuth cathodes. The rate of the reduction of nitrate was about the same on both metals. The selectivity (%S) to ammonia was similar on the two metals (12% at 450 mA/cm(2)) and that to nitrogen 82% on Sn and 72% on Bi. On the other hand, the %S to nitrous oxide was lower on Sn (8%) than that on Bi (18%) under the same conditions. The current efficiency (%CE) on both metals was 80% when the 99% of the initial nitrogen was removed. The %CE gradually decreases as the concentration of the nitrogen containing species in the solution decreases during the electrolysis. The energy consumption for the removal of 1g of N was 100 Wh at 450 mA/cm(2) but it can be significantly reduced by a better design of the electrolysis cell.The presence of chromate in the electrolyte, which is a known inhibitor of the cathodic reduction of nitrate, has no influence on the rate of the reduction and the distribution of the products. PMID- 19559524 TI - Stripping/flocculation/membrane bioreactor/reverse osmosis treatment of municipal landfill leachate. AB - This study presents a configuration for the complete treatment of landfill leachate with high organic and ammonium concentrations. Ammonia stripping is performed to overcome the ammonia toxicity to aerobic microorganisms. By coagulation-flocculation process, COD and suspended solids (SS) were removed 36 and 46%, respectively. After pretreatment, an aerobic/anoxic membrane bioreactor (Aer/An MBR) accomplished the COD and total inorganic nitrogen (total-N(i)) removals above 90 and 92%, respectively, at SRT of 30 days. Concentrations of COD and total-N(i) (not considering organic nitrogen) in the Aer/An MBR effluent decreased to 450 and 40 mg/l, respectively, by significant organic oxidation and nitrification/denitrification processes. As an advanced treatment for the leachate, the reverse osmosis (RO) was applied to the collected Aer/An MBR effluents. Reverse osmosis provided high quality effluent by reducing the effluent COD from MBR to less than 4.0mg/l at SRT of 30 days. PMID- 19559525 TI - Characterization of wet precipitation by X-ray diffraction (XRD) and scanning electron microscopy (SEM) in the metropolitan area of Porto Alegre, Brazil. AB - The purpose of this study is to assess the composition of wet precipitation in three sites of the metropolitan area of Porto Alegre. Besides the variables usually considered, such as pH, conductivity, major ions (Cl(-), NO(3)(-), F(-), SO(4)(2-), Na(+), K(+), Mg(2+), NH(4)(+) and Ca(2+)) and metallic elements (Cd, Co, Cr, Cu, Fe, Mn and Ni), the suspended matter was examined by X-ray diffraction (XRD) and scanning electron microscopy (SEM), with energy dispersive system (EDS), for better identification of possible anthropogenic material in this wet precipitation. Results showed an alkaline pH in the samples analyzed and higher concentrations for Na(+), Cl(-) and SO(4)(2-). The acidification and neutralization potential between anions (SO(4)(2-)+NO(3)(-)) and cations (Ca(2+)+Mg(2+)+K(+)+NH(4)(+)) showed a good correlation (0.922). The metallic elements with highest values were Zn, Fe and Mn. Results of XRD identified the presence of some minerals such as quartz, feldspar, mica, clay, carbonates and sulfates. In samples analyzed with SEM, we detected pyroxene, biotite, amphibole and oxides. Cluster analysis (CA) was applied to the data matrix to identify potential pollution sources of metals (natural or anthropogenic) and the association with minerals found in the analysis of SEM. PMID- 19559526 TI - Hg(II) removal from aqueous solution by bayberry tannin-immobilized collagen fiber. AB - A novel adsorbent was prepared by immobilizing barberry tannin (BT) onto collagen fiber, which was found effective to remove Hg(II) from aqueous solution. The bayberry tannin-immobilized collagen fiber (BTICF) shows high adsorption capacity to Hg(II) in a wide pH range of 4.0-9.0, and a maximum adsorption capacity (198.49 mg/g) was reached at pH 7.0 and 303 K when the initial concentration of Hg(II) was 200.0 mg/L. The adsorption isothermal and kinetic data were well fitted by the Langmuir equation and the pseudo-first-order rate equation, respectively. The adsorption mechanism of BTICF to Hg(II) was proved to follow a chelating reaction. The BTICF can be easily regenerated with 0.1M lactic acid after adsorption process and recycled at least 4 times without the loss of adsorption capacity. These facts indicate that BTICF can be used as a low-cost adsorbent for effective removal of Hg(II) from aqueous solutions. PMID- 19559527 TI - Controllable solvothermal synthesis and photocatalytic properties of complex (oxy)fluorides K2TiOF4, K3TiOF5, K7Ti4O4F7 and K2TiF6. AB - Complex (oxy)fluorides K(2)TiF(6), K(2)TiOF(4), K(3)TiOF(5) and K(7)Ti(4)O(4)F(7) have been successfully synthesized for the first time through a controllable solvothermal route involving different solvents, for example, methanol, methanol H(2)O and methanol-H(2)O(2). The as-prepared products were characterized by X-ray powder diffraction, N(2) surface area adsorption, scanning electron microscope, Fourier transform infrared spectroscopy, UV-vis absorption spectra and X-ray fluorescence. The influences of reaction conditions such as the ratio of methanol to H(2)O(2) or methanol to H(2)O, reaction temperature on the phase, crystallizability and purity of the (oxy)fluorides products were discussed in detail. Meanwhile, the photocatalytic behaviors of the as-prepared K(2)TiF(6), K(2)TiOF(4), K(3)TiOF(5) and K(7)Ti(4)O(4)F(7) were evaluated by degradation of rhodamine B molecules, and the results showed that all of the products possessed photocatalytic activities in the order of K(2)TiOF(4)>K(2)TiF(6)>K(7)Ti(4)O(4)F(7)>K(3)TiOF(5) at room temperature under the UV light. PMID- 19559528 TI - Open your mind to placebo conditioning. PMID- 19559529 TI - Factors contributing to large analgesic effects in placebo mechanism studies conducted between 2002 and 2007. AB - Recent meta-analyses find various magnitudes of placebo analgesia effects in placebo mechanism trials versus placebo control trials, which have led to debate. To further investigate the magnitude of placebo analgesia in placebo mechanism trials the databases "PubMed", "PsycINFO" and "Web of Science" (2002-2007) were searched with the term "placebo analgesia". Twenty-one articles including 24 studies fulfilled the selection criteria (concerning: mechanisms, control, placebo treatment, randomization and pain measures). The validity of studies was assessed by the authors and effect sizes were calculated via difference scores. The magnitude of placebo analgesia in placebo mechanism studies was large (d=1.00) and about five times larger than placebo analgesia effects in placebo control studies (d=0.15-0.27). Differences in magnitude between the two types of studies appear to result from different types of suggestions given for pain relief. The magnitude of placebo effects was larger in studies that used long term pain stimuli >20s (d=0.96) as opposed to short-term stimuli (d=0.81) and the largest placebo effects were found in studies wherein hyperalgesia was present (d=1.88). These results replicate our previous finding that placebo analgesic effects are higher in mechanism studies than in placebo control studies. However, since magnitudes of placebo analgesic effects are highly variable it may be valuable to investigate the factors and mechanisms that contribute to this variability as well as differences in magnitudes across types of studies. PMID- 19559530 TI - Influence of the tip mass and position on the AFM cantilever dynamics: coupling between bending, torsion and flexural modes. AB - The effects of the geometrical asymmetric related to tip position as a concentrated mass, on the sensitivity of all three vibration modes, lateral excitation (LE), torsional resonance (TR) and vertical excitation (VE), of an atomic force microscopy (AFM) microcantilever have been analyzed. The effects of the tip mass and its position are studied to report the novel results to estimating the vibration behavior of AFM such as resonance frequency and amplitude of the microcantilever. In this way, to achieve more accurate results, the coupled motion in all three modes is considered. In particular, it is investigated that performing the coupled motion in analysis of AFM microcantilever is almost necessary. It is shown that the tip mass and its position have significant effects on vibrational responses. The results show that considering the tip mass decreases the resonance frequencies particularly on high order modes. However, dislocating of tip position has an inverse effect that causes an increase in the resonance frequencies. In addition, it has been shown that the amplitude of the AFM microcantilever is affected by the influences of tip and its position. These effects are caused by the interaction between flexural and torsional motion due to the moment of inertia of the tip mass. PMID- 19559531 TI - Toxoplasmosis in pigs--the last 20 years. AB - Pigs are important to the economy of many countries because they are a source of food for humans. Infected pig meat is a source of Toxoplasma gondii infection for humans and animals in many countries. This parasite also causes mortality in pigs, especially neonatal pigs. Most pigs acquire T. gondii infection postnatally by ingestion of oocysts from contaminated environment or ingestion of infected tissues of animals. Few pigs become infected prenatally by transplacental transmission of the parasite. Raising pigs indoors in confinement has greatly reduced T. gondii infection in pigs but the recent trend of organic farming is likely to increase T. gondii infection in pigs. Recently, feeding goat whey to pigs was found to be a risk factor for T. gondii infection in organically raised pigs. Currently used molecular and histopathological methods are insensitive for the detection of T. gondii in pork because of the low concentration of the parasite in meat destined for human consumption. There is no vaccine to prevent T. gondii infection in pigs but efforts are being continued to develop a non viable vaccine. In the present paper, information on prevalence, transmission, diagnosis, and control of porcine toxoplasmosis in the last 20 years (since 1988 when last reviewed by this author) is reviewed. Worldwide reports of clinical and asymptomatic infections in pigs are reviewed. Methods to detect T. gondii in pigs are compared. Recent studies on genetic typing of T. gondii strains prevalent in pigs are discussed with respect to epidemiology. Because wild pigs are hunted for food for human consumption prevalence in wild pigs is summarized. PMID- 19559532 TI - Monocyte- and macrophage-mediated immune reactions against Eimeria bovis. AB - Innate immune reactions conducted by macrophages may affect the outcome of primary infections and are crucial for the transition to adaptive immune responses. In bovine coccidiosis little is known on early monocyte/macrophage mediated responses. We therefore investigated in vivo, in vitro and ex vivo reactions of monocytes and macrophages against Eimeria bovis, one of the most pathogenic Eimeria species in cattle. Macrophages significantly infiltrate the gut mucosa of E. bovis-infected calves, particularly after challenge infection. Furthermore, peripheral monocytes of infected animals, as precursor cells of macrophages, exhibited enhanced ex vivo phagocytic and oxidative burst activities. Enhanced levels of both activities were found early after infection and towards the end of first merogony. In vitro exposure of macrophages to sporozoites led to phagocytosis of the pathogen, whilst monocytes failed to do so. Phagocytosis occurred independently of the viability of the sporozoites, indicating that active invasion by the parasites was negligible. Phagocytosis occurred in the absence of immune serum, but could clearly be enhanced by addition of immune serum, suggesting macrophage-derived antibody-dependent cytotoxicity. Furthermore, co-culture of macrophages with sporozoites and stimulation with merozoite I antigen induced distinct levels of cytokine and chemokine gene transcription. Thus, the transcription of genes encoding for IFN gamma, IL-12, TNF-alpha, IL-6, CXCL1, CXCL8, CXCL10 and COX-2 was upregulated after sporozoite encounter. In contrast, soluble merozoite I antigen only induced the gene transcription of IL-6 and IL-12 and failed to upregulate IFN-gamma and TNF-alpha gene transcripts. In monocytes, IFN-gamma and CXCL10 were found upregulated, all other immunoregulatory molecules tested were not affected. In summary, our results strongly suggest that macrophage-mediated, innate immune reactions play an important role in the early immune response to E. bovis infections in calves. PMID- 19559533 TI - Experimental transmission of Hepatozoon americanum to New Zealand White rabbits (Oryctolagus cuniculus) and infectivity of cystozoites for a dog. AB - Inflammatory lesions containing parasitic cystozoites developed in multiple organs and tissues of laboratory-raised Oryctolagus cuniculus that were administered approximately 100 sporulated oocysts of Hepatozoon americanum (Oklahoma isolate, GenBank accession AF176836) orally. The predominantly granulomatous inflammatory lesions were detected histologically 8 weeks after exposure to oocysts. Cystozoites, recognized by cresent-shaped, uninucleated bodies surrounded by an accumulation of globular, PAS-positive polysaccharide material, were evident within macrophages as monozoic and dizoic cysts. Neither meronts nor gamonts were detected in any of the laboratory-raised lagomorphs during the 24-week observation period. Nested PCR assay of rabbit tissues for a 488 bp fragment of the 18S rRNA Hepatozoon spp. gene was positive at 8 and 24 weeks post-exposure. The sequence was 97.1% similar with sequence from the H. americanum carrier used to infect ticks. A Hepatozoon-free dog fed tissues from the 24-week post-exposure rabbit principal developed American canine hepatozoonosis. Gamonts were first detected 5 weeks after the dog ingested the rabbit tissues containing cystozoites. PCR assay of blood from the dog was positive for the Hepatozoon spp. gene fragment. Sequencing confirmed that the parasite in the dog was H. americanum. PMID- 19559534 TI - Clinical and biochemical studies on Theileria annulata in Egyptian buffaloes (Bubalus bubalis) with particular orientation to oxidative stress and ketosis relationship. AB - This study was carried out on 68 Theileria annulata naturally infected buffaloes in addition to 25 parasitologically free buffaloes distributed in small herds at Dakahlia and Gharbya governorates, Egypt, to demonstrate the clinical picture associated with theileriosis in this buffaloes with particular emphasis to the oxidative stress and ketosis relationship. Clinical signs recorded in infected buffaloes were in the form of fever, enlargement of one or more lymph node, ocular discharge, corneal opacity, skin lesions, decreased milk yield, pale mucous membrane and anorexia. Blood and serum analysis revealed significant (p or =7.0). The lower education degree, injection, poly drug use and 3 or more relapse were dependently associated with the severe nicotine dependence, and the adjusted odds ratio (OR) were 3.8 (1.5-10.0), 2.3 (1.3-4.0), 3.7 (2.1-6.5) and 1.9 (1.1-3.4) respectively. This study exhibited high rate of cigarette smoking in Chinese opiate addicts and the majority had severe nicotine dependence and suggested that a comprehensive intervention of cigarette smoking should be paid attention to during substance abuse treatment in China. PMID- 19559540 TI - Reducing the risk of skin pathologies in diabetics by using copper impregnated socks. AB - Diabetic individuals frequently suffer from skin pathologies, especially in their feet. Co-existing peripheral vascular disease and neuropathy exacerbate the capacity of these individuals to cope with infections, minor cuts and wounds, often leading to hard to treat and chronic ulcers. Copper has potent anti-fungal and antibacterial properties. Copper is also an essential trace element vital for the normal function of many tissues and indispensable for the generation of new capillaries and skin. Human skin is not sensitive to copper and the risk of adverse reactions due to dermal exposure to copper is extremely low. We hypothesize that part of the increased risk of developing foot skin pathologies in diabetic patients with compromised blood circulation to the foot is due to low local copper levels. We further hypothesize that copper ions released from copper impregnated socks and absorbed through the skin would improve the well-being of the skin of diabetic patients by inducing angiogenesis and expression and stabilization of extracellular skin proteins, in addition to their biocidal effect of reducing the risk of fungal and bacterial infection of the diabetic foot. Thus, the use of copper impregnated socks may be used as a preventive modality. Furthermore, we hypothesize that the copper released from the socks may even be beneficial in the healing of cuts, wounds and even hard to treat skin pathologies. PMID- 19559541 TI - Could we prevent unilateral cleft lip/palate in the future? AB - Surveillance studies have shown that cleft lip and palate is one of the commonest craniofacial anomalies, occurring in approximately 1 in 500 live births. Previous studies on craniofacial form in unilateral cleft lip/palate subjects have been carried out, but most attention has been focused on the deformity of the bony septum whereas the deformities of the nasal spine and cartilaginous component of the septum had received little attention. Our recent study was based on monitoring a very specific type of nasal septal deformity, type 6, and its relation to the unilateral cleft lip/palate disease. This type is very anteriorly located and refers to the cartilaginous part of the nasal septum and the inter maxillary bone itself. Rhinoscopic view shows a typical, almost horizontal, unilateral groove at the nasal septum located very anteriorly. At the opposite septal side, but corresponding location, there is so called basal crest. The results of our study showed that the incidence of type 6 septal deformity was very high not only in unilateral cleft lip/palate children (80.6%) but also in their parents (58% in at least one of them). In contrast, in our previous study this type of septal deformity was seen in only 3.7% of non-unilateral cleft lip/palate children before puberty, rising to 7.4% in students and 9.4% in adults. In other words, perhaps we can expect the onset of unilateral cleft lip/palate in the offspring of parents who both have a type 6 septal deformity. Perhaps there is a gene responsible for the onset of both type 6 septal deformity and the cleft. If these clinical entities belong to the same gene, the cleft per se could perhaps disappear from the Earth in a near future owing to the gene therapy which will be able to eliminate it before the baby is born or even conceived. PMID- 19559542 TI - [Cystadenofibroma of the ovary: report of two cases]. AB - Cystadenofibroma of the ovary is a relatively rare benign tumor. Such tumors are characterised by their malignant macroscopical appearance which may lead to an inappropriate aggressive surgical approach. We present two cases of cystadenofibromas of the ovary. The first has been treated by extensive surgery, including pelvic and para-aortic lymphadenectomy. The second has been treated in a more appropriate way, by conservative surgery. PMID- 19559543 TI - Young adult injection drug users in the United States continue to practice HIV risk behaviors. AB - BACKGROUND: Injection drug users (IDUs) are at risk of acquiring HIV through injection and sexual practices. METHODS: We analyzed data collected in five U.S. cities between 2002 and 2004 to identify correlates of HIV infection among 3285 IDUs ages 15-30 years. RESULTS: Overall, HIV prevalence was 2.8% (95% CI 2.3 3.4), ranging from 0.8% in Chicago to 6.3% in Los Angeles. Mean age was 24 years, 70% were male, 64% non-Hispanic (NH) white, 7% NH black, 17% Hispanic, and 12% were other/mixed race. HIV infection was independently associated with: race/ethnicity (NH black [AOR 4.1, 95% CI 1.9-9.1], Hispanic [AOR 3.6, 95% CI 1.5 8.4], or other/mixed [AOR 2.3, 95% CI 1.1-5.2] vs. NH white); males who only had sex with males compared to males who only had sex with females (AOR 15.3, 95% CI 6.8-34.5); injecting methamphetamine alone or with heroin compared to heroin only (AOR 4.0, 95% CI 1.7-9.7); reporting inconsistent means of obtaining income compared to regular jobs (AOR 2.3, 95% CI 1.1-4.8); and having a history of exchanging sex for money/drugs (AOR 2.8, 95% CI 1.5-5.2). CONCLUSIONS: More than two decades after injection and sexual practices were identified as risk factors for HIV infection, these behaviors remain common among young IDUs. While racial/ethnic disparities persist, methamphetamine may be replacing cocaine as the drug most associated with HIV seropositivity. HIV prevention interventions targeting young IDUs and address both sexual and injection practices are needed. PMID- 19559544 TI - Prodynorphin gene deficiency potentiates nalbuphine-induced behavioral sensitization and withdrawal syndrome in mice. AB - Dynorphin is the presumed endogenous ligand for the kappa-opioid receptor. The dynorphin gene may play a role in psychotropic agent-mediated behavioral changes via dopaminergic modulation. Therefore, in this study, possible involvement of the dynorphin gene in nalbuphine-mediated behavioral responses was examined using prodynorphin (Pdyn) gene knock-out (-/-) mice. Pdyn gene deficiency potentiates nalbuphine-induced behavioral sensitization of locomotor activity and accumbal c Fos expression. Administration of nalbuphine induced a significant increase in the dialysate dopamine level in the nucleus accumbens. This increase was more pronounced in the Pdyn (-/-) mice than in the wild-type (WT) mice. In addition, Pdyn (-/-) mice were more vulnerable to the naloxone-precipitated withdrawal syndrome (i.e., teeth chattering, wet dog shakes, forepaw tremors, jumping, weight loss, and global withdrawal score) after repeated treatment with nalbuphine than the WT mice. Consistently, nor-binaltorphimine, a kappa-opioid receptor antagonist, significantly potentiated nalbuphine-induced behavioral effects in WT mice, whereas U-50488H, a kappa-opioid receptor agonist, significantly attenuated these changes in Pdyn (-/-) mice in a dose-dependent manner. Our data suggest that the kappa-opioid receptor/dynorphin system is specifically modulated in response to behavioral sensitization and withdrawal signs induced by nalbuphine. PMID- 19559545 TI - Dynamics of plc gene transcription and alpha-toxin production during growth of Clostridium perfringens strains with contrasting alpha-toxin production. AB - The aim of the present study was to investigate transcription dynamics of the alpha-toxin-encoding plc gene relative to two housekeeping genes (gyrA and rplL) in batch cultures of three Clostridium perfringens strains with low, intermediate, and high levels of alpha-toxin production, respectively. The plc transcript level was always low in the low alpha-toxin producing strain. For the two other strains, plc transcription showed an inducible pattern and reached a maximum level in the late exponential growth phase. The transcription levels were however inversely correlated to alpha-toxin production for the two strains. We propose that this discrepancy is due to differences in plc translation rates between the strains and that strain-specific translational rates therefore must be determined before alpha-toxin production can be extrapolated from transcript levels in C. perfringens. PMID- 19559546 TI - Evaluation of isolation procedures and chromogenic agar media for the detection of MRSA in nasal swabs from pigs and veal calves. AB - Since the emergence of MRSA in livestock, screening of animals for the detection of MRSA is widely practised. Different procedures are published for animal samples but a systematic comparison of methods has not been performed. The objective of this study was to compare three available commonly used procedures and three chromogenic agars for detecting MRSA in nasal swabs from pigs (n=70) and veal calves (n=100). Procedures 1 and 2 used a pre-enrichment comprising Mueller Hinton broth with 6.5% NaCl followed by selective enrichment with 4 microg/ml oxacillin+75 microg/ml aztreonam (procedure 1) and 5 microg/ml ceftizoxime+75 microg/ml aztreonam (procedure 2) respectively. Procedure 3 used a selective enrichment broth only, containing 4% NaCl, 5 microg/ml ceftizoxime+50 microg/ml aztreonam. After selective enrichment, media were streaked on to three different chromogenic agars. Significantly more MRSA were found for pig as well as for veal calf samples with procedures 1 and 2. No significant differences were found between procedures 1 and 2. For nasal swabs from pigs significantly more MRSA-positive samples were found when MRSA Screen (Oxoid) or MRSASelect (Bio-Rad) agars were used compared to MSRA ID (bioMerieux). For calf samples no significant differences between the different agars were found. In conclusion, the results of this study show that procedures 1 and 2, both using additional high salt pre enrichment are superior and should be recommended for MRSA detection in nasal swabs from pigs and veal calves. The preferred choice of chromogenic agar depends on the sample matrix. PMID- 19559547 TI - A PCR method of detecting American Foulbrood (Paenibacillus larvae) in winter beehive wax debris. AB - The objective of this work was to create a fast and sensitive method of detecting Paenibacillus larvae from beehive debris based on PCR that does not require long lasting cultivation steps. Various methods of extracting spores from beehive debris were compared: the original method of extraction of spores into toluene, and alternative spore extraction methods into Tween 80, into water, into isopropanol and into 95% ethanol. Isolation of DNA from various spore extractions was evaluated too. Best results were provided by isolation of DNA using the QIAamp DNA Mini Kit, without heat treatment. DNA of spores was detected by PCR from 0.25 g of beeswax debris, with the detected titer of 10(5) in 1g according to the cultivation tests. PMID- 19559548 TI - Swine torque teno virus (TTV) infection and excretion dynamics in conventional pig farms. AB - Torque teno virus (TTV) is a non-enveloped, single-stranded DNA (ssDNA) virus infecting human and non-primate species. Two genogroups of TTV (TTV1 and TTV2) have been described in swine so far. In the present study, TTV1 and TTV2 prevalences in serum, and nasal as well as rectal swabs of 55 randomly selected piglets from seven Spanish multi-site farms, were monitored from 1 to 15 weeks of age. Also, blood from their dams (n=41) were taken at 1 week post-farrowing. Samples were tested by means of two TTV genogroup specific PCRs. Although prevalence of TTV1 and TTV2 in sows was relatively high (54% and 32%, respectively), it was not directly associated to their prevalence in the offspring. Percentage of viremic pigs for both TTV genogroups followed similar dynamics, increasing progressively over time, with the highest rate of detection at 11 weeks of age for TTV1 and at 15 weeks for TTV2. Forty-two (76%) and 33 (60%) of the 55 studied pigs were TTV1 and TTV2 PCR positive in serum, respectively, in more than one sampling time. TTV1 and TTV2 viremia lasted in a number of animals up to 15 and 8 weeks, respectively. Co-infection with both TTV genogroups in serum was detected at all sampling points, but at 1 week of age. On the contrary, there were animals PCR negative to both genogroups in serum at all sampling times but at 15 weeks of age. During the study period, TTV1 and TTV2 nasal shedding increased also over time and faecal excretion was intermittent and of low percentage (<20%). In conclusion, the present study describes for the first time the infection dynamics of TTV1 and TTV2 as well as the nasal and faecal excretion throughout the life of pigs from conventional, multi-site farms. Moreover, results indicate that both swine TTV genogroups are able to establish persistent infections in a number of pigs. PMID- 19559550 TI - Quiz--An ossified soft tissue mass. PMID- 19559549 TI - How many CT detector rows are necessary to perform adequate three dimensional visualization? AB - INTRODUCTION: The technical development of computer tomography (CT) imaging has experienced great progress. As consequence, CT data to be used for 3D visualization is not only based on 4 row CTs and 16 row CTs but also on 64 row CTs, respectively. The main goal of this study was to examine whether the increased amount of CT detector rows is correlated with improved quality of the 3D images. MATERIAL AND METHODS: All CTs were acquired during routinely performed preoperative evaluation. Overall, there were 12 data sets based on 4 detector row CT, 12 data sets based on 16 detector row CT, and 10 data sets based on 64 detector row CT. Imaging data sets were transferred to the DKFZ Heidelberg using the CHILI teleradiology system. For the analysis all CT scans were examined in a blinded fashion, i.e. both the name of the patient as well as the name of the CT brand were erased. For analysis, the time for segmentation of liver, both portal and hepatic veins as well as the branching depth of portal veins and hepatic veins was recorded automatically. In addition, all results were validated in a blinded fashion based on given quality index. RESULTS: Segmentation of the liver was performed in significantly shorter time (p<0.01, Kruskal-Wallis test) in the 16 row CT (median 479 s) compared to 4 row CT (median 611 s), and 64 row CT (median 670 s), respectively. The branching depth of the portal vein did not differ significantly among the 3 different data sets (p=0.37, Kruskal-Wallis test). However, the branching depth of the hepatic veins was significantly better (p=0.028, Kruskal-Wallis test) in the 4 row CT and 16 row CT compared to 64 row CT. The grading of the quality index was not statistically different for portal veins and hepatic veins (p=0.80, Kruskal-Wallis test). Even though the total quality index was better for the vessel tree based on 64 row CT data sets (mean scale 2.6) compared to 4 CT row data (mean scale 3.25) and 16 row CT data (mean scale 3.0), these differences did not reach statistical difference (p=0.53, Kruskal-Wallis test). CONCLUSION: Even though 3D visualization is useful in operation planning, the quality of the 3D images appears to be not dependent of the number of CT detector rows. PMID- 19559551 TI - Contrast-enhanced ultrasound is helpful in the differentiation of malignant and benign breast lesions. AB - OBJECTIVE: To evaluate the significance of contrast-enhanced ultrasound (CEUS) examination in differential diagnosis of malignant and benign breast lesions. METHODS: Seventy-one patients with seventy-six breast tumors are selected randomly. CEUS examinations were performed before and after bolus injection of the contrast agent SonoVue (Bracco, Milan, Italy). Specific sonographic quantification software, Qontrast, was adopted to determine the morphology of vessels. Wash-in and wash-out parameters of each lesion were assessed for both procedures. RESULTS: The final histopathological findings distinguished 45 malignant and 31 benign from all of the lesions. Following SonoVue administration different perfusion phases could be identified: early (0-1min), mid (1-4min) and late (4-6min) phases. In the early phase, CEUS identified 91.1% of malignant tumors characterized by a claw-shaped enhancement, while 83.9% of benign tumors had a homogeneous enhancement, with a statistically significant difference between the two enhancement patterns (chi(2)=43.16, P<0.01). Moreover, contrast medium persistence in the late phase was helpful in the identification of benign and malignant tumors (chi(2)=46.88, P<0.01): contrast medium was present in 88.9% of malignant tumors, while in only 9.7% of the benign tumors. The study showed that various parametric imaging color maps for peak intensity and time to peak were mostly suggestive of malignancy, while quite uniform peak intensity and time to peak of color maps were the characteristic of benign tumors. The study also found that malignant lesions presented with a higher maximum intensity signal than benign ones (P<0.05) on the time-intensity curves. CONCLUSIONS: CEUS cooperating with conventional US shows improved accuracy in differentiating between malignant and benign breast tumors. It could be a reliable diagnostic method of breast lesions. PMID- 19559552 TI - MRI-based biomechanical imaging: initial study on early plaque progression and vessel remodeling. AB - The goal of the study is to develop a noninvasive magnetic resonance imaging (MRI)-based biomechanical imaging technique to address biomechanical pathways of atherosclerotic progression and regression in vivo using a 3D fluid-structure interaction (FSI) model. Initial in vivo study was carried out in an early plaque model in pigs that underwent balloon-overstretch injury to the left carotid arteries. Consecutive MRI scans were performed while the pigs were maintained on high cholesterol (progression) or normal chow (regression), with an injection of a plaque-targeted contrast agent, Gadofluorine M. At the end of study, the specimens of carotid arterial segments were dissected and underwent dedicated mechanical testing to determine their material properties. 3D FSI computational model was applied to calculate structure stress and strain distribution. The plaque structure resembles early plaque with thickened intima. Lower maximal flow shear stress correlates with the growth of plaque volume during progression, but not during regression. In contrast, maximal principle structure stress/stain (stress-P1 and strain-P1) were shown to correlate strongly with the change in the plaque dimension during regression, but moderately during progression. This MRI based biomechanical imaging method may allow for noninvasive dynamic assessment of local hemodynamic forces on the development of atherosclerotic plaques in vivo. PMID- 19559553 TI - Contrast-enhanced MR angiography with frequency-dependent mask subtraction. AB - One of the main challenges for high-resolution contrast-enhanced magnetic resonance angiography (CE-MRA) is the limited signal-to-noise ratio (SNR). In conventional CE-MRA, spoiled gradient-recalled echo sequence is typically used to acquire raw data before and after contrast material injection, followed by mask subtraction to improve vessel contrast at the cost of increased image noise. We reported here on a frequency-dependent mask subtraction technique where only low spatial frequency data were subject to mask subtraction to improve image contrast and reduce motion artifact while high spatial frequency data were not subtracted to preserve image noise level. The feasibility of this technique was demonstrated through phantom, animal, and human volunteer studies. The lowest half, quarter, 1/8 and 1/16 of all spatial frequencies were subject to mask subtraction, respectively. These partial subtraction techniques were compared with conventional mask subtraction in terms of SNR and artifact power. An SNR gain up to 37% was achieved without significant artifact power increase for quarter subtraction where only the central one quarter k-space data were subject to mask subtraction. PMID- 19559554 TI - An optimized solenoidal head radiofrequency coil for low-field magnetic resonance imaging. AB - Applications of low-field magnetic resonance imaging (MRI) systems (<0.3 T) are limited due to the signal-to-noise ratio (SNR) being lower than that provided by systems based on superconductive magnets (> or = 1.5 T). Therefore, the design of radiofrequency (RF) coils for low-field MRI requires careful consideration as significant gains in SNR can be achieved with the proper design of the RF coil. This article describes an analytical method for the optimization of solenoidal coils. Coil and sample losses are analyzed to provide maximum SNR and optimum B(1) field homogeneity. The calculations are performed for solenoidal coils optimized for the human head at 0.2 T, but the method could also be applied to any solenoidal coil for imaging other anatomical regions at low field. Several coils were constructed to compare experimental and theoretical results. A head magnetic resonance image obtained at 0.2 T with the optimum design is presented. PMID- 19559555 TI - Retrospective correction for T1-weighting bias in T2 values obtained with various spectroscopic spin-echo acquisition schemes. AB - Localized tissue transverse relaxation time (T(2)) is obtained by fitting a decaying exponential to the signals from several spin-echo experiments at different echo times (TE). Unfortunately, time constraints in magnetic resonance spectroscopy (MRS) often mandate in vivo acquisition schemes at short repetition times (TR), that is, comparable with the longitudinal relaxation constant (T(1)). This leads to different T(1)-weighting of the signals at each TE. Unaccounted for, this varying weighting causes systematic underestimation of the T(2)'s, sometimes by as mush as 30%. In this article, we (i) analyze the phenomenon for common MRS spin-echo T(2) acquisition schemes; (ii) propose a general post hoc T(1)-bias correction for any (TR, TE) combination; (iii) show that approximate knowledge of T(1) is sufficient, since a 20% uncertainty in T(1) leads to under 3% bias in T(2); and consequently, (iv) efficient, precision-optimized short TR spin-echo T(2) measurement protocols can be designed and used without risk of accuracy loss. Tables of correction for single-refocusing (conventional) spin echo and double refocusing, such as, PRESS acquisitions, are provided. PMID- 19559557 TI - A serial functional connectivity MRI study in healthy individuals assessing the variability of connectivity measures: reduced interhemispheric connectivity in the motor network during continuous performance. AB - To date, little data is available on the reproducibility of functional connectivity MRI (fcMRI) studies. Here, we tested the variability and reproducibility of both the functional connectivity itself and different statistical methods to analyze this phenomenon. In the main part of our study, we repeatedly examined two healthy subjects in 10 sessions over 6 months with fcMRI. Cortical areas involved in motor function were examined under two different cognitive states: during continuous performance (CP) of a flexion/extension task of the fingers of the right hand and while subjects were at rest. Connectivity to left primary motor cortex (lSM1) was calculated by correlation analysis. The resulting correlation coefficients were transformed to z-scores of the standard normal distribution. For each subject, multisession statistical analyses were carried out with the z-score maps of the resting state (RS) and the CP experiments. First, voxel based t tests between the two groups of fcMRI experiments were performed. Second, ROI analyses were carried out for contralateral right SM1 and for supplementary motor area (SMA). For both ROI, mean and maximum z-score were calculated for each experiment. Also, the fraction of significantly (P<.05) correlated voxels (FCV) in each ROI was calculated. To evaluate the differences between the RS and the CP condition, paired t tests were performed for the mean and maximum z-scores, and Wilcoxon signed ranks tests for matched pairs were carried out for the FCV. All statistical methods and connectivity measures under investigation yielded a distinct loss in left-right SM1 connectivity under the CP condition. For SMA, interindividual differences were apparent. We therefore repeated the fcMRI experiments and the ROI analyses in a group of seven healthy subjects (including the two subjects of the main study). In this substudy, we were able to verify the reduction of left-right SM1 connectivity during unilateral performance. Still, the direction of SMA to lSM1 connectivity change during the CP condition remained undefined as four subjects showed a connectivity increase and three showed a decrease. In summary, we were able to demonstrate a distinct reduction in left-right SM1 synchrony in the CP condition compared to the RS both in the longitudinal and in the multisubject study. This effect was reproducible with all statistical methods and all measures of connectivity under investigation. We conclude that despite intra- and interindividual variability, serial and cross-sectional assessment of functional connectivity reveals stable and reliable results. PMID- 19559556 TI - Dynamic contrast-enhanced magnetic resonance imaging parameters independent of baseline T10 values. AB - A baseline T(10) value is typically needed for dynamic contrast-enhanced (DCE-) MRI studies. However, an assumed baseline T(10) has to be used when T(10) measurements for patients are not available. In this work, we systematically investigate the dependence on T(10) of the commonly used DCE-MRI parameters (K(trans), k(ep), v(e) and IAUC) as well as several newly defined parameters [the normalized ratios (NRs) of k(ep), K(trans) and v(e), which are measures of relative changes in these parameters between two time points] for a spoiled gradient-echo pulse sequence using simulations and in vivo studies. Effects of various factors on the T(10) dependence, such as the true T(10) value, flip angle and the potential changes in T(10) due to treatment, were also assessed using simulations. We found that DCE-MRI parameters k(ep) and the NR of k(ep) are largely independent of T(10), especially when larger flip angles are used (e.g., 30-40 degrees). Their estimations therefore do not require any knowledge of T(10). The NRs of K(trans), v(e) and IAUC also exhibit independence to T(10), but only when T(10) remains constant between pre- and posttreatment. The estimation of parameters themselves (K(trans), v(e) and IAUC) is highly dependent on the T(10) value. PMID- 19559558 TI - Detection and grading for esophageal varices in patients with chronic liver damage: comparison of gadolinium-enhanced and unenhanced steady-state coherent MR images. AB - The purpose of this study was to compare observer interpreted steady-state coherent coronal images and gadolinium-enhanced axial images in terms of the detection and grading of esophageal varices. Magnetic resonance imaging (MRI) and gastrointestinal endoscopy were performed within 2 weeks in 90 patients with chronic liver damage, including 55 with untreated esophageal varices, for periodic screening purposes. Two blinded readers retrospectively reviewed T1- and T2-weighted images with gadolinium-enhanced (gadolinium image set) and steady state coherent (coherent image set) images. Sensitivity for the detection of esophageal varices was higher (P<.001) in the gadolinium image set (76%) than in the coherent image set (35%); on the other hand, specificity was higher (P<.001) in the coherent image set (91%) than in the gadolinium image set (66%). Furthermore, area under the ROC curve was higher for the gadolinium image set (Az=0.823) than the coherent image set (Az=0.761) (P=.48). Moderate and weak positive correlations with endoscopic grades were found for the gadolinium image (r=0.48, P<.01) and coherent image sets (r=0.34, P=.018). The addition of steady state coherent imaging to the current routine liver imaging protocol did not improve the detection or grading of esophageal varices, whereas gadolinium enhanced imaging was found to be potentially valuable. Nevertheless, endoscopy was confirmed to be mandatory in patients with esophageal varices suspected by MRI of the liver. PMID- 19559559 TI - Rotating frame relaxation during adiabatic pulses vs. conventional spin lock: simulations and experimental results at 4 T. AB - Spin relaxation taking place during radiofrequency (RF) irradiation can be assessed by measuring the longitudinal and transverse rotating frame relaxation rate constants (R(1rho) and R(2rho)). These relaxation parameters can be altered by utilizing different settings of the RF irradiation, thus providing a useful tool to generate contrast in MRI. In this work, we investigate the dependencies of R(1rho) and R(2rho) due to dipolar interactions and anisochronous exchange (i.e., exchange between spins with different chemical shift deltaomega not equal0) on the properties of conventional spin-lock and adiabatic pulses, with particular emphasis on the latter ones which were not fully described previously. The results of simulations based on relaxation theory provide a foundation for formulating practical considerations for in vivo applications of rotating frame relaxation methods. Rotating frame relaxation measurements obtained from phantoms and from the human brain at 4 T are presented to confirm the theoretical predictions. PMID- 19559560 TI - Estradiol-17beta and linseed meal interact to alter visceral organ mass and hormone concentrations from ovariectomized ewes. AB - To evaluate the estrogenic potential of secoisolariciresinol diglycoside (SDG) found in linseed meal (LSM) on visceral organ mass, IGF-I, and thyroid hormone (T(3) and T(4)) concentrations, 48 multiparous, ovariectomized ewes (54.6 +/- 1.1 kg) were used in a 3 x 4 factorial arrangement. Main effects were length of LSM feeding (0, 1, 7, or 14 d) and length of exposure to estradiol-17beta (E(2)) implant (0, 6, or 24 h prior to tissue collection). Implanting ewes with E(2) for 24h increased liver mass relative to empty body weight (EBW; g/kg EBW) compared with ewes implanted for 0 or 6 h (P or= 0.12) CYP2C or CYP3A mRNA expression or cellularity of the liver. Exogenous E(2) influenced circulating concentrations of IGF-I, T(3), and T(4). The estrogenic or anti-estrogenic potential of LSM is dependent upon the tissue, exposure to E(2), and the duration of LSM feeding. Feeding LSM during gestation, lactation, or during the grow finish phase warrants further investigation. PMID- 19559561 TI - Factors affecting bony impingement in hip arthroplasty. AB - Computer modeling of 10 patients' computed tomographic scans was used to study the variables affecting hip arthroplasty range of motion before bony impingement (ROMBI) including acetabular offset and height, femoral offset, height and anteversion, and osteophyte removal. The ROMBI was compared with the ROM before component impingement and the native hip ROM. The ROMBI decreased with decreased total offset and limb shortening. Acetabular offset and height had a greater effect on ROMBI than femoral offset and height. The ROMBI lost with decreased acetabular offset was not fully recoverable with an increase in femoral offset or osteophyte removal. Bony impingement increased and component impingement decreased with decreased acetabular offset and increased head diameter. PMID- 19559562 TI - Variability in femoral component rotation reference axes measured during navigation-assisted total knee arthroplasty using gap technique. AB - The basic objective in total knee arthroplasty is to achieve the correct amount of femoral component rotation, and this can be achieved either with a measured resection technique or indirectly with a flexion/extension gap equalization technique. We studied variability in the reference axes (posterior condylar axis, Whiteside's line, transepicondylar axis) when soft tissue tension was managed intraoperatively using a navigation system. The mean angles for the transepicondylar line, Whiteside's line, and the posterior condylar line from the proximal tibia resection plane were 1.29 degrees +/- 3.67 degrees (mean +/- SD; range, -7 degrees to 10.5 degrees), 3.90 degrees +/- 4.17 degrees (mean +/- SD; range, -3 degrees to 15.5 degrees), and -4.03 degrees +/- 2.71 degrees (mean +/- SD; range, -9.5 degrees to 1.0 degrees), respectively. The coefficients of variation (SD/mean x100) for these 3 variables were 283%, 106%, and 67%, respectively. Of the 3 reference axes used widely for femoral component rotation, the angles from the posterior condylar line to the proximal tibia resection plane showed the smallest range of variance. PMID- 19559563 TI - Exploration of concerns of relatives during continuous palliative sedation of their family members with cancer. AB - Data on the experiences of relatives during continuous palliative sedation are scarce. Because these relatives may be the ones most closely involved with the patient, it is important to evaluate the possible burdens that they experience. We aimed to explore and evaluate concerns of relatives during continuous palliative sedation of their family members admitted to an acute palliative care unit. Through retrospective multidisciplinary record research, we obtained data on concerns of the relatives during the period that continuous palliative sedation took place. From October 2001 to October 2004, 45 patients died after starting continuous palliative sedation. In 51% of the cases, the relatives expressed concerns after starting the therapy. Concerns could be distinguished into three main themes: concerns about the aim of continuous palliative sedation (27%), concerns related to the well-being of the patient (29%), and concerns related to the well-being of relatives themselves (18%). Patient and sedation characteristics did not differ significantly between sedations in which relatives did and did not express concerns, except for the duration of the sedation. The median duration of the continuous palliative sedation when concerns were expressed was 46 hours, compared with 19.5 hours when this was not the case (P<0.05). Both the nature and extent of the concerns suggest that relatives are in need of continuous information and professional guidance during continuous palliative sedation of their family members. Availability of caregiver guidance and clear process documentation are crucial and indispensable in providing this. PMID- 19559564 TI - Delivering palliative care in an acute hospital setting: views of referrers and specialist providers. AB - There has been a steady expansion of hospital-based palliative care in the United Kingdom but limited published research on health professionals' views of hospital multidisciplinary specialist palliative care services (SPCS). The aim of the study was to describe referrer (SPCS user) and provider (SPCS staff) perspectives on delivery of specialist palliative care in hospital. Interviews were conducted with referrers, including five junior doctors, 13 consultants, and six clinical nurse specialists, to investigate the reasons for referral, beneficial aspects, and barriers to use. Focus groups were conducted with providers, six medical and five nursing, to identify their perspective on delivering the specialist service in hospital. Discussions were tape recorded and transcribed verbatim. Data were analyzed thematically using a framework analysis approach. The study found large areas of agreement between referrers and providers on what hospital palliative care teams should be providing for patients, that is, expertise in managing difficult symptoms and complex psychosocial problems, and this was being achieved locally. Access to the specialist team was also important: visibility on the wards, informal routes of access to advice and a timely response by specialists. However, discordance in views of providing palliative care was also identified; in particular, whether specialists should be providing generalist palliative care (such as basic psychological support) neglected by ward teams and implementation of specialist advice by referrers. Such perspectives on the interface of generalist and specialist provision provide insights into improving care for palliative patients in the acute hospital setting. PMID- 19559565 TI - Noninvasive ventilation for the treatment of dyspnea as a bridge from intensive to end-of-life care. PMID- 19559566 TI - Incidence and evolution of mural thrombus in abdominal aortic endografts. AB - BACKGROUND: The aims of this study were to analyze the predictive factors for intragraft mural thrombus formation and evolution during follow-up after endovascular treatment of abdominal aortic aneurysms and to evaluate its relationship with the subsequent appearance of complications. METHODS: A retrospective study was performed by selecting those patients who underwent endovascular repair of an abdominal aortoiliac aneurysm between June 1998 and September 2004, with a minimum follow-up of 24 months. Preoperative clinical data, anatomical characteristics of the aneurysm, and endograft type were analyzed. In addition, clinical evaluation and abdominal computed tomography angiography (CT scans) performed at 1, 6, 12, and 24 months after the surgery were reviewed. RESULTS: Eighty-nine patients were submitted for endovascular aneurysm repair in this period, and 75 completed the 24-month follow-up. Eighteen patients developed intragraft mural thrombus (24% incidence), 13 (72.2%) appearing at 1 month of follow-up, and up to 16 (88.9%) appearing during the first 6 months. Logistic regression analysis showed that the lumen percentage of mural thrombus in the native aorta and the use of aortouniiliac endografts were independent predictors of intragraft mural thrombus formation (odds ratio, 1.065; 95% confidence interval, 1.022-1.110, and odds ratio, 8.014; 95% confidence interval, 1.598-40.181, respectively). No spontaneous regression of the thrombus was observed. The area of intragraft mural thrombus had increased at 12 and 18-24 months after their diagnosis (Wilcoxon signed rank test, p = 0.028 and 0.028, respectively). The presence of intragraft mural thrombus was associated with a greater tendency to endograft body or branch occlusion (5 of 18 cases with intragraft mural thrombus (27.8%) versus 1 of 57 cases without it (1.8%), (p = 0.003). CONCLUSION: Intragraft formation of mural thrombus is a common finding during the follow-up of abdominal aortic endografts, particularly in aneurysms with large mural thrombus of the native aorta, and is associated with the use of aortouniiliac endografts. The area occupied by the mural thrombus was shown to gradually increase during follow-up and was associated with a greater tendency for endograft occlusion. PMID- 19559567 TI - Endovascular treatment of a blunt aortic injury in Iraq: extension of innovative endovascular capabilities to the modern battlefield. AB - BACKGROUND: The management of blunt descending thoracic aortic injury remains controversial. Despite emerging evidence touting the advantage of endovascular repair in civilian trauma, there have been no reports on the application of this management strategy in the austere environment of war. We provide a case report from the 332nd EMDG/Air Force Theater Hospital, Balad Air Base, Iraq. METHODS: An Iraqi policeman presented with traumatic aortic disruption following blunt trauma. The patient arrived with hemoperitoneum, a Le Fort III facial fracture, a left humerus fracture, and a thoracic aortic disruption. Following facial packing, fracture stabilization, and damage control laparotomy, aortography was performed, confirming aortic disruption beyond the left subclavian artery. The injury was treated with three aortic cuffs (Gore Excluder AAA Aortic Extender Endoprostheses) placed in sequence from the origin of the left subclavian across the disruption. A type III endoleak was successfully managed with placement of one additional aortic cuff. RESULTS: The patient was discharged after 1 month following the successful treatment of his other injuries. Computed tomography angiography at 10 and 30 days following the procedure revealed no endoleak and a resolved periaortic hematoma. CONCLUSION: This report details the first endovascular treatment of blunt aortic injury in wartime and represents a sustained commitment to advance innovative endovascular capability closer to the time of injury. Although controversial, this less invasive approach is appealing in patients with high injury severity scores, making its availability in wartime especially germane. PMID- 19559568 TI - Management of antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is characterised by vascular thrombosis and/or obstetric morbidity in the presence of persistently positive antiphospholipid antibodies (aPL). Balancing an individuals' risk of thrombosis against the benefits and risks of antithrombotic therapies is crucial for optimising management and preventing morbidity in APS and asymptomatic aPL. Limitations in research studies have led to debate regarding best-practice. This review of the available literature makes the following recommendations. Those with asymptomatic aPL should only be treated with aspirin if they have persistently positive aPL, obstetric APS, or co-existent systemic lupus erythematosus. For those with APS, lower risk patients (i.e. first venous thrombosis) should be treated with warfarin to an INR 2.0-3.0. Those at higher risk (i.e. arterial thrombosis or recurrent events) should be treated with warfarin to an INR of >3.0. During pregnancy in APS, low molecular weight heparin (LMWH) and aspirin should be used and women should be under the care of obstetricians and physicians specialising in APS. Additional vascular and thrombotic risk factors should be actively reduced in all patient groups. Further randomised controlled trials are required, which should involve larger patient groups with APS diagnosed according to accepted criteria. This may mean that international and multi-centre trials are needed to ascertain the best treatment regimens. PMID- 19559569 TI - Mechanical anisotropy of orthodontic mini-implants. AB - This study investigated stress distribution in the bone around orthodontic mini implants using the finite element method and determined the difference in the stress distribution for different loading directions to identify risk factors for the loosening of mini-implants. Three-dimensional finite element models were constructed for conventional and cervical threadless mini-implants with cortical bone 1 or 3mm thick. The authors calculated the compressive stresses on the bone elements and evaluated stress distribution according to the loading direction. Directional dependency (i.e. mechanical anisotropy) was observed with the conventional mini-implant model. The compressive stress ranged from -31 to -55 MPa depending on the loading direction. In the cervical threadless model, mechanical anisotropy disappeared and the stress was reduced. Cortical bone thickness had no influence in either model. One of the risk factors for mini implant failure might be related to mechanical anisotropy. This report suggests ways for clinicians to avoid overload traction force when conventional mini implants are used. The cervical threadless mini-implant can reduce mechanical anisotropy to facilitate successful placement. Inserting a conventional screw deeply beyond the threaded part might be useful in stabilizing a mini-implant. PMID- 19559570 TI - Superior temporal gray and white matter changes in schizophrenia or antipsychotic related effects? PMID- 19559571 TI - HB-EGF-induced VEGF production and eNOS activation depend on both PI3 kinase and MAP kinase in HaCaT cells. AB - BACKGROUND: Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of growth factors that have been implicated in skin patho-physiology. Although endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) appear to be involved in mitogenesis and chemotaxis in epidermal keratinocytes, the activation of eNOS and VEGF production induced by HB-EGF and its signaling mechanism remains undefined. OBJECTIVE: We examined possible signal transduction pathways by which HB-EGF leads to eNOS activation and VEGF production in human epidermal keratinocyte cell line (HaCaT cells). METHODS: The phosphorylation of epidermal growth factor receptor (EGFR), mitogen-activated protein kinase (MAPK; p42/p44 MAPK), Akt and eNOS were examined by Western blotting analysis. VEGF production was determined by enzyme-linked immunosorbent assay. Various inhibitors were utilized to investigate the signaling mechanisms of eNOS activation and VEGF production. RESULTS: HB-EGF-induced phosphorylation of EGFR with maximum phosphorylation at 1h. HB-EGF-induced phosphorylation of p42/p44 MAPK in a few minutes. It activated Akt with maximum phosphorylation at 1h and eNOS with maximum phosphorylation at 3h. The HB-EGF-induced eNOS activation was significantly blocked by the p42/p44 MAPK inhibitor U0126 and the phosphatidylinositol 3-kinase (P13K) inhibitor LY294002. HB-EGF increased VEGF production. The HB-EGF-induced VEGF production was blocked by U0126 and LY294002. Finally, the HB-EGF-induced activation of Akt and eNOS was suppressed by VEGF competitive antagonist, CBO-P11. CONCLUSION: These results demonstrate that HB EGF-induced eNOS activation depends on p42/p44 MAPK, PI3K/Akt pathways and endogenous VEGF in HaCaT cells. PMID- 19559572 TI - Early improvement as a predictor of remission and response in schizophrenia: Results from a naturalistic study. AB - OBJECTIVE: To examine the predictive validity of early improvement in a naturalistic sample of inpatients and to identify the criterion that best defines early improvement. METHODS: Two hundred and forty-seven inpatients who fulfilled ICD-10 criteria for schizophrenia were assessed with the Positive And Negative Syndrome Scale (PANSS) at admission and at biweekly intervals until discharge from hospital. Remission was defined according to the recently proposed consensus criteria, response as a reduction of at least 40% in the PANNS total score from admission to discharge. RESULTS: Receiver operating characteristic (ROC) analyses showed that early improvement (reduction of the PANSS total score within the first 2 weeks of treatment) predicts remission (AUC=0.659) and response (AUC=0.737) at discharge. A 20% reduction in the PANSS total score within the first 2 weeks was the most accurate cut-off for the prediction of remission (total accuracy: 65%; sensitivity: 53%; specificity: 76%), and a 30% reduction the most accurate cut-off for the prediction of response (total accuracy: 76%; sensitivity: 47%; specificity: 90%). CONCLUSION: The findings of clinical drug trials that early improvement is a predictor of subsequent treatment response were replicated in a naturalistic sample. Further studies should examine whether patients without early improvement benefit from an early change of antipsychotic medication. PMID- 19559574 TI - Association between cerebral metabolic and structural abnormalities and cognitive performance in schizophrenia. AB - The possible association in schizophrenia between frontal abnormalities, such as hypofrontality and frontal grey matter (GM) deficits, and neuropsychological deficits is not yet well defined. Our objective was to study such an association and to clarify the cognitive relevance of metabolic and anatomical variability across schizophrenia patients. To do so, we studied dorsolateral prefrontal (DLPF) metabolism during an attention test using fluoro-deoxy-glucose positron emission tomography and DLPF structure with magnetic resonance imaging (MRI) in 22 schizophrenia patients [9 neuroleptic-naive (NN) first episodes]. These patients also underwent a comprehensive battery of neuropsychological tests aimed at evaluating global intelligence and the proposed domains of cognitive alteration in schizophrenia, i.e., attention, visual and verbal learning and memory, working memory, problem solving and processing speed. The metabolic activity in the right DLPF region was significantly and directly related to processing speed, and a measure of structural deficit in the same area was directly related to working memory scores. In the NN group studied alone, these associations were replicated. We may conclude that hypofrontality during cognitive activation, and the degree of DLPF structural deficit may be associated to a particular profile of cognitive deficit, including lower processing speed and working memory capacity. PMID- 19559573 TI - Subcortical volumetric correlates of anxiety in familial pediatric bipolar disorder: a preliminary investigation. AB - Anxiety is a common comorbid condition in pediatric bipolar disorder (BD). However, there is little known about the effects of comorbidity on brain morphometry in this population. The aim of the present study was to examine subcortical correlates of anxiety in familial pediatric BD. The subject group comprised 120 children (mean age=12+/-3.3 years) with at least one parent diagnosed with BD. Bipolar offspring with BD were compared with bipolar offspring without BD on a measure of overall lifetime anxiety. A sub-sample of 20 bipolar offspring with BD (mean age=14.6+/-2.8 years) underwent magnetic resonance imaging (MRI) with a 3-T scanner. Correlational analyses were conducted between hippocampal and amygdalar volumes, and anxiety scores. The results showed significantly higher anxiety in bipolar offspring with BD compared to bipolar offspring without BD. There was a significant negative association between total hippocampal volume and anxiety scores. No significant association was found between total amygdalar volume and anxiety scores. Clinically, these findings suggest that anxiety comorbidity needs to be properly assessed and treated in the management of pediatric BD. This is the first study to show a negative association between hippocampal volume and anxiety in this population. The overlap between anxiety and familial pediatric BD suggests that anxiety may be one important area of future research in parsing out the heterogeneous nature and complex etiology of early-onset BD. PMID- 19559576 TI - Surface bio-modification of poly(hydroxybutyrate-co-hydroxyhexanoate) and its aging effect. AB - In this work, poly(hydroxybutyrate-co-hydroxyhexanoate) (PHBHHx) film was fabricated by a solution-casting method and subsequently was modified by NaOH treatment to improve the surface hydrophilic property. Surface properties including hydrophilicity, surface appearance and functional groups were characterized by water contact angle measurement, scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The results showed the hydrophilicity of PHBHHx film was obviously improved by the NaOH treatment due to the topography changes promoted by the NaOH-etching and the introduction of polar groups included hydroxyl and carboxyl on the topmost surface layers. However, the modified film exhibited an aging effect: the hydrophilicity decreases with time elapsed during storage. It was found that the aging rate was strongly dependent on the crystallinity of the film and the storage environment. The sample with high-crystallinity lost hydrophilic property slower than that with low crystallinity. Hydrophilic and low-temperature environment also prevented the modified PHBHHx from fast losing of the hydrophilicity. PMID- 19559575 TI - The relation of worry to prefrontal cortex volume in older adults with and without generalized anxiety disorder. AB - Despite the widespread prevalence of generalized anxiety disorder (GAD) in later life, almost nothing is known about the neural aspects of worry in adults over the age of 60. Given the ongoing rapid increase in the older adult population, the relatively poor response rates to current interventions for late life GAD, and the effects of age-related changes to the brain, additional research on worry neurobiology is needed. The study group comprised 15 older GAD patients and 15 matched controls who were compared on clinical measures and brain volumes. It was expected that prefrontal cortex (PFC) volumes [medial orbital cortex (mOFC), dorsolateral cortex (DLPFC)] would show positive relations to worry scores, and weaker relations to more general measures of anxiety and depression. Negative relations were expected between amygdala volumes and worry scores. As expected, mOFC volumes were positively related to worry scores; however, DLPFC and amygdala volumes were not. The mOFC is involved in emotional decision-making under uncertain conditions and has the ability to suppress the amygdala, both of which are hypothesized functions of worry. Results are partly consistent with GAD theory and suggest that worry may involve neural areas that are also involved in the successful control of anxiety. PMID- 19559578 TI - Covalent attachment of trypsin on plasma polymerized allylamine. AB - This paper focuses on the immobilization of a proteolytic enzyme, trypsin, on plasma polymerized allylamine (ppAA) films. The later have been deposited onto silicon substrate by means of radiofrequency glow discharge. The covalent attachment of the enzyme was achieved in three steps: (i) activation of the polymer surface with glutaraldehyde (GA) as a linker, (ii) immobilization of trypsin and (iii) imino groups reduction treatment. The effects and efficiency of each step were investigated by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). Fluorescent spectroscopy was used to evaluate the change of the biological activity following the immobilization steps. The results showed that enzyme immobilization on GA-modified substrate increases the enzyme activity by 50% comparing to adsorbed enzymes, while the imino reduction treatment improves the enzyme retention by about 30% comparing to untreated samples. In agreement with XPS and AFM data, UV-vis absorption spectroscopy, used to quantify the amount of immobilized enzyme, showed that allylamine plasma polymer presents a high adsorption yield of trypsin. Although the adsorbed enzymes exhibit a lower activity than that measured for enzymes grafted through GA linkers, the highest catalytic activity obtained was for the enzymes that underwent the three steps of the immobilization process. PMID- 19559577 TI - Capillary force required to detach micron-sized particles from solid surfaces- validation with bubbles circulating in water and 2 microm-diameter latex spheres. AB - The adhesion forces holding micron-sized particles to solid surfaces can be studied through the detachment forces developed by the transit of an air-liquid interface in a capillary. Two key variables affect the direction and magnitude of the capillary detachment force: (i) the thickness of the liquid film between the bubble and the capillary walls, and (ii) the effective angle of the triple phase contact between the particles and the interface. Variations in film thickness were calculated using a two-phase flow model. Film thickness was used to determine the time-variation of the capillary force during transit of the bubble. The curve for particle detachment was predicted from the calculated force. This curve proved to be non-linear and gave in situ information on the effective contact angle developing at the particle-bubble interface during detachment. This approach allowed an accurate determination of the detachment force. This theoretical approach was validated using latex particles 2 microm in diameter. PMID- 19559579 TI - Biomacromolecular affinity: interactions between lysozyme and regioselectively sulfated chitosan. AB - It is well known that the sulfate groups on different positions in polysaccharides play important roles in protein adsorption. However, the interactions between sulfated chitosans and lysozyme have not been clearly elucidated. In this study, the regioselectively sulfated chitosans, 6-O-sulfated chitosan (C6S), 2-N-6-O-sulfated chitosan (C26S) and 3,6-O-sulfated chitosan (C36S), were chosen to investigate the possible mechanisms determining the interaction between lysozyme and the sulfated chitosans. It has been found that the selectively sulfated products of chitosan (CS), C6S, C26S and C36S all exhibit lysozyme binding activity. However, the maximum binding ratios of lysozyme/polysaccharide are significantly different for C6S, C26S and C36S. In addition, though C6S possesses the lowest sulfur content among the three sulfated chitosans, it exhibits the highest binding activity with lysozyme. Furthermore, in the protein mixtures, C6S shows the highest selective binding activity with lysozyme among the three sulfated chitosans in the presence of gamma-globulin and bovine serum albumin (BSA). The results indicate that 6-O-sulfate groups may be responsible for the high affinity and specific interaction of sulfated chitosan with lysozyme, while 2-O-sulfate and 3-O-sulfate groups are unfavorable to this interaction. PMID- 19559580 TI - Nano- and macro-geometrical structural change of caffeine and theophylline anhydrate tablets during hydration process by using X-ray computed tomography. AB - The effects of nano- and macro-geometrical factors on the hydration kinetics of caffeine (CA) and theophylline anhydrate (TA) tablets at high humidity were investigated using X-ray high-resolution computed tomography (CT). Hydration profiles of CA and TA tablets obtained at 25 and 50 MPa, 96% relative humidity, and 20 degrees C were measured by weight and X-ray CT. The total tablet volume (G V) and average tablet density (G-D) calculated based on the volume and weight of tablets, and tablet volume (CT-V) and tablet density (CT-D) were evaluated by X ray CT. The hydration kinetics of CA and TA tablets followed two-dimensional growth of nuclei (Avrami-Erofee) and three-dimensional phase boundary equations, respectively. The increase in the G-V of TA tablets was initially more than, but later less than, that of CA tablets. The G-D of CA tablets varied extensively and was constant initially, whereas that of TA tablets decreased significantly in the initial stage. The CT-V of CA tablets gradually increased initially, but that of TA tablets increased significantly early on. The inter-granular volume (IG-V) of both tablets decreased initially with large fluctuations, but then increased. The CT-D of CA tablets decreased significantly, but that of TA tablets did not decrease. The hydration kinetics of CA and TA tablets was affected by changes in the geometrical structure of the tablets. X-ray CT is a powerful tool for evaluating dynamic changes inside tablets. PMID- 19559581 TI - Studies on the kinetics of growth of silver nanoparticles in different surfactant solutions. AB - Silver nanoparticles were prepared in aqueous silver nitrate solution using hydrazine as reducing agents in presence of two ionic surfactants (cetyltrimethylammonium bromide; CTAB and sodium dodecyl sulfate; SDS) and one non-ionic surfactant (Triton X-100). The reaction rate was determined spectrophotometrically. The nature of the head group of these surfactants is responsible for the formation of stable, yellow and transparent silver sol. For a certain reaction time, i.e., 20 min, the absorbance of reaction mixture first increased until it reached a maximum, then decreased with [hydrazine]. The reaction follows first-order kinetics with respect to each in [hydrazine] and [Ag(+)]. The results suggest formation of a complex between silver(I) and hydrazine, decomposes in a rate-determining step, leading in the formation of a free radical, which again reacts with the silver(I) in a subsequent fast step to yield the products. The transmission electron microscopic (TEM) images show that CTAB stabilized silver nanoparticles are spherical and of uniform particle size, and the average particle size is about 15 nm. PMID- 19559582 TI - Morphometric analysis of brain images with reduced number of statistical tests: a study on the gender-related differentiation of the corpus callosum. AB - OBJECTIVE: We evaluate the feasibility of applying dynamic recursive partitioning (DRP), an image analysis technique, to perform morphometric analysis. We apply DRP to detect and characterize discriminative morphometric characteristics between anatomical brain structures from different groups of subjects. Our method reduces the number of statistical tests, commonly required by pixel-wise statistics, alleviating the effect of the multiple comparison problem. METHODS AND MATERIALS: The main idea of DRP is to partition the two-dimensional (2D) image adaptively into progressively smaller subregions until statistically significant discriminative regions are detected. The partitioning process is guided by statistical tests applied on groups of pixels. By performing statistical tests on groups of pixels rather than on individual pixels, the number of statistical tests is effectively reduced. This reduction of statistical tests restricts the effect of the multiple comparison problem (i.e., type-I error). We demonstrate an application of DRP for detecting gender-related morphometric differentiation of the corpus callosum. DRP was applied to template deformation fields computed from registered magnetic resonance images of the corpus callosum to detect regions of significant expansion or contraction between female and male subjects. RESULTS: DRP was able to detect regions comparable to those of pixel-wise analysis, while reducing the number of required statistical tests up to almost 50%. The detected regions were in agreement with findings previously reported in the literature. Statistically significant discriminative morphological variability was detected in the posterior corpus callosum region, the isthmus and the anterior corpus callosum. In addition, by operating on groups of pixels, DRP appears to be less prone to detecting spatially diffused and isolated outlier pixels as significant. CONCLUSION: DRP can be a viable approach for detecting discriminative morphometric characteristics among groups of subjects, having the potential to alleviate the multiple comparisons' effect by significantly reducing the number of required statistical tests. PMID- 19559583 TI - Toxic effects of prolonged administration of leaves of cassava (Manihot esculenta Crantz) to goats. AB - Cassava (Manihot esculenta Crantz) is a major source of dietary energy for humans and domestic animals in many tropical countries. However, consumption of cassava is limited by its characteristic content of cyanogenic glycosides. The present work aimed to evaluate the toxic effects of ingestion of cassava leaves by goats for 30 consecutive days, and to compare the results with the toxic effects of cyanide in goats, which have been described previously. Eight Alpine cross-bred female goats were divided into two equal groups, and were treated with ground frozen cassava leaves at a target dose of 6.0mg hydrogen cyanide (HCN)/kg/day (treated animals), or with ground hay and water only (control group) by gavage for 30 consecutive days. Blood samples were collected on days 0, 7, 15, 21, and 30 for biochemical panel and cyanide determination. At the end of the experiment, fragments of pancreas, thyroid gland, liver, kidney, lungs, heart, spleen, and the whole central nervous system were collected for histopathological examination. Clinical signs were observed in all goats treated with cassava on the first day of the experiment. From the second day the dose of cassava leaves was reduced to 4.5mgHCN/kg/day. No changes were found in the blood chemical panel. A mild increase in the number of resorption vacuoles in the thyroid follicular colloid, slight vacuolation of periportal hepatocytes, and spongiosis of the mesencephalon were found in goats treated with cassava. The pattern of lesions seen in the present goats was similar to what has been described previously in cyanide-dosed goats. Thus, the toxic effects of the ingestion of cassava leaves by goats can be attributed to the action of cyanide released from cyanogenic glycosides, and none of the effects was promoted by these glycosides directly. PMID- 19559584 TI - Cryo-electron tomography in biology and medicine. AB - During the last six decades electron microscopy (EM) has been essential to ultra structural studies of the cell to understand the fundamentals of cellular morphology and processes underlying diseases. More recently, electron tomography (ET) has emerged as a novel approach able to provide three-dimensional (3D) information on cells and tissues at molecular level. Electron tomography is comparable to medical tomographic techniques like CAT, PET and MRI in the sense that it provides a 3D view of an object, yet it does so at a cellular scale and with nanometer resolution. Electron tomography has the unique ability to visualize molecular assemblies, cytoskeletal elements and organelles within cells. The three-dimensional perspective it provides has revised our understanding of cellular organization and its relation with morphological changes in normal development and disease. Cryo-electron tomography of vitrified samples at cryogenic temperatures combines excellent structural preservation with direct high-resolution imaging. The use of cryo-preparation and imaging techniques eliminates artifacts induced by plastic embedding and staining of the samples is circumvented. This review describes the technique of cryo-electron tomography, its basic principles, cryo-specimen preparation, tomographic data acquisition and image processing. A number of illustrative examples ranging from whole cells, cytoskeletal filaments, viruses and organelles are presented along with a comprehensive list of research articles employing cryo-electron tomography as the key ultrastuctural technique. PMID- 19559585 TI - Human innate immunity against African trypanosomes. AB - Humans are naturally resistant to infection by the African trypanosome prototype Trypanosoma brucei brucei, and only two variant clones of this parasite can avoid this innate immunity and cause sleeping sickness. The resistance to T. brucei is due to serum complexes associating apolipoprotein A-1 (apoA1) with two primate specific proteins, apolipoprotein L-1 (apoL1) and haptoglobin-related protein (Hpr). We discuss recent advances on the respective functions of apoL1 and Hpr in this system. ApoL1 was found to share structural and functional similarities with proteins of the apoptotic Bcl2 family, and to kill trypanosomes through anionic pore formation in the lysosomal membrane of the parasite. In association with hemoglobin (Hb), Hpr was found to promote the binding of the trypanolytic complexes to a haptoglobin (Hp)-Hb receptor of the trypanosome surface, hereby facilitating the internalization of apoL1. Hpr or apoL1 deficiency respectively leads to the reduction or abolishment of human protection against T. brucei. PMID- 19559586 TI - Membranes and organelles. PMID- 19559587 TI - The construction of a glucose-sensing luciferase. AB - A novel luminescence-based glucose-sensing molecule was created by combining a galactose-/glucose-binding protein (GGBP) with luciferase. The glucose-sensing luciferase (GlcLuc) was constructed using a GGBP fused with a large domain and a small domain of Firefly luciferase (Lluc and Sluc). The luminescence intensity based analysis with E. coli recombinant protein showed that the GlcLuc had luciferase activity in glucose or galactose in a concentration-dependent manner (K(d)=3.9 microM for glucose and 11 microM for galactose), and that the increase in the activity saturated within one minute after the injection of the ligands. These results indicated that the conformation change of the GGBP moiety following the ligand binding effectively induced the reconstitution of the GGBP-fused split luciferase. The Asp459Asn mutation, which was expected to lead to a glucose specific binding ability, was then introduced into the GlcLuc. The GlcLuc mutant showed the luciferase activity increasing only with the increase of glucose concentration, but not with that of galactose. Our results demonstrate that the GGBP fused with a split luciferase, which is reconstituted rapidly and specifically in the presence of glucose, provides a novel glucose-sensing system based on luminescence and may also contribute to the construction of luminescence based sensing molecules for other substrates using other PBPs. PMID- 19559588 TI - Polycrystalline bismuth oxide films for development of amperometric biosensor for phenolic compounds. AB - An attractive biocomposite based on polycrystalline bismuth oxide (BiO(x)) film and polyphenol oxidase (PPO) was proposed for the construction of a mediator-free amperometric biosensor for phenolic compounds in environmental water samples. The phenolic biosensor could be easily achieved by casting the biocomposite on the surface of glassy carbon electrode (GCE) via the cross-linking step by glutaraldehyde. The laboratory-prepared bismuth oxide semiconductor was polymorphism. Its hydrophilicity provided a favorable microenvironment for retaining the biological activity of the immobilized protein. The parameters of the fabrication process and the various experimental variables for the enzyme electrode were optimized. The proposed PPO/BiO(x) biosensor provided a linear response to catechol over a concentration range of 4 x 10(-9)M to 1.5 x 10(-5)M with a dramatically developed sensitivity of 11.3 AM(-1)cm(-2) and a detection limit of 1 x 10(-9)M based on S/N=3. In addition, the PPO/BiO(x) biocomposite was characterized by scanning electron microscope (SEM), Fourier transform infrared spectra (FTIR) and rotating disk electrode voltammetry. PMID- 19559589 TI - Individually addressable microelectrode arrays fabricated with gold-coated pencil graphite particles for multiplexed and high sensitive impedance immunoassays. AB - A renewable, site-selective immobilization platform of microelectrode array (MEA) for multiplexed immunoassays has been initially developed using pencil graphite particles coated with gold layers as microelectrodes. The graphite particles available on the common pencil were utilized for directing the electro-deposition of gold layers with uniform microstructures which displayed a well-defined sigmoidal voltammetric response. In the concept-of-proof experiments, the resulting MEA platform was modified with functionalized monolayer, on which anti human IgG antibodies could be stably immobilized in a site-selective way through binding chemistry to selectively capture human IgG antigens from the sample media. The subsequent introduction of anti-human IgG antibodies conjugated with 15 nm electro-active gold nanoparticles to recognize the captured IgG proteins resulted in a significant decrease in the interfacial electron-transfer resistance. High sensitive electrochemical quantification by gold nanoparticle amplified impedance responses could thus be achieved. Experimental results show that the developed MEA sensor can allow for the detection of human IgG with wide linear range (0.05-100 ng ml(-1)) and sensitivity over 10(3) larger than that of the conventional, bulk gold electrode. The rapid regeneration of the used MEA platform can additionally be realized by a simple electrochemical treatment. The high selectivity of four individually addressable MEA platforms for multiple antigens in a single sample has been further demonstrated in the multiplexed immunoassay experiments. Such a site-selective immobilization strategy of MEA platform may open a new door towards the development of various simple, sensitive, cost-effective, and reusable biological sensors and biochips. PMID- 19559590 TI - Label-free and sensitive faradic impedance aptasensor for the determination of lysozyme based on target-induced aptamer displacement. AB - A label-free and sensitive faradic impedance spectroscopy (FIS) aptasensor based on target-induced aptamer displacement was developed for the determination of lysozyme as a model system. The aptasensor was fabricated by self-assembling the partial complementary single strand DNA (pcDNA)-lysozyme binding aptamer (LBA) duplex on the surface of a gold electrode. To measure lysozyme, the change in interfacial electron transfer resistance of the aptasensor using a redox couple of [Fe(CN)(6)](3-/4-) as the probe was monitored. The introduction of target lysozyme induced the displacement of the LBA from the pcDNA-LBA duplex on the electrode into the solution, decreasing the electron transfer resistance of the aptasensor. The decrease in the FIS signal is linear with the concentration of lysozyme in the range from 0.2 nM to 4.0 nM, with a detection limit of 0.07 nM. The fabricated aptasensor shows a high sensitivity, good selectivity and satisfactory regeneration. This work demonstrates that a high sensitivity of the fabricated aptasensor can be obtained using a relatively short pcDNA. This work also demonstrates that the target-induced aptamer displacement strategy is promising in the design of an electrochemical aptasensor for the determination of lysozyme with good selectivity and high sensitivity. PMID- 19559591 TI - Three-dimensional focusing of red blood cells in microchannel flows for bio sensing applications. AB - Three-dimensional (3D) focusing of particles in microchannels has been a long standing issue in the design of biochemical/biomedical microdevices. Current microdevices for 3D cell or bioparticle focusing involve complex channel geometries in view of their fabrication because they require multiple layers and/or sheath flows. This paper proposes a simple method for 3D focusing of red blood cells (RBCs) in a single circular microcapillary, without any sheath flows, which is inspired from the fluid dynamics phenomenon in that a spherical particle lagging behind a Poiseuille flow migrates toward and along the channel axis. More explicitly, electrophoresis of RBCs superimposed on the pressure-driven flow is utilized to generate an RBC migration mode analogous to this phenomenon. A particle-tracking scheme with a sub-pixel resolution is implemented to spatially position red blood cells flowing through the channel, so that a probability density function (PDF) is constructed to evaluate the tightness of the cell focusing. Above a specific strength of the electric field, approximately 90% of the sheep RBCs laden in the flow are tightly focused within a beam diameter that is three times the cell dimension. Particle shape effect on the focusing is discussed by making comparisons between the RBCs and the spherical particles. The lateral migration velocity, predicted by an existing theoretical model, is in good agreement with the present experimental data. It is noteworthy that 3D focusing of non-spherical particles, such as RBCs, has been achieved in a circular microchannel, which is a significant improvement over previous focusing methodologies. PMID- 19559592 TI - Real-time monitoring of odorant-induced cellular reactions using surface plasmon resonance. AB - Surface plasmon resonance (SPR) is a powerful technique for measuring molecular interaction in real-time. SPR can be used to detect molecule to cell interactions as well as molecule to molecule interactions. In this study, the SPR-based biosensing technique was applied to real-time monitoring of odorant-induced cellular reactions. An olfactory receptor, OR I7, was fused with a rho-tag import sequence at the N-terminus of OR I7, and expressed on the surface of human embryonic kidney (HEK)-293 cells. These cells were then immobilized on a SPR sensor chip. The intensity of the SPR response was linearly dependent on the amount of injected odorant. Among all the aldehyde containing odorants tested, the SPR response was specifically high for octanal, which is the known cognate odorant for the OR I7. This SPR response is believed to have resulted from intracellular signaling triggered by the binding of odorant molecules to the olfactory receptors expressed on the cell surface. This SPR system combined with olfactory receptor-expressed cells provides a new olfactory biosensor system for selective and quantitative detection of volatile compounds. PMID- 19559593 TI - Development of molecularly imprinted polymers for the binding of nitrofurantoin. AB - Novel molecularly imprinted polymers (MIPs) for the recognition of nitrofurantoin (NFT) were prepared by photoinitiated polymerisation in polar solvent using 2,6 bis(methacrylamido) pyridine (BMP) as the functional monomer and carboxyphenyl aminohydantoin (CPAH) as the analogue of the template. The binding constants of the complex between BMP and nitrofurantoin or CPAH in DMSO were determined with 1H NMR titration to be 630+/-104 and 830+/-146 M(-1), respectively. To study the influence of the functional monomer, two polymer compositions were prepared containing the template, the functional monomer and the crosslinker in the molar ratio 1:1:12 for MIP1 and 1:4:20 for MIP2, respectively. The imprinting factor at saturation concentration of nitrofurantoin, which is the ratio of the amount bound to the MIP and the non-imprinted control polymer (NIP), was determined to be 2.47 for MIP1 and 2.49 for MIP2. The cross reactivity of the imprinted polymers seems to be determined by the ability to form hydrogen bonds to the functional monomer while the shape of the molecule has no real influence. PMID- 19559594 TI - Crystal violet-G-quadruplex complexes as fluorescent sensors for homogeneous detection of potassium ion. AB - A novel K(+) detection method was reported using a label-free G-quadruplex forming oligonucleotide and a triphenylmethane fluorescent dye crystal violet (CV). This method is based on the fluorescence difference of some CV/G-quadruplex complexes in the presence of K(+) or Na(+), and the fluorescence change with the variation of K(+) concentration. According to the nature of the fluorescence change of CV as a function of ionic conditions, two K(+) detection modes can be developed. One is a fluorescence-decreasing mode, in which T(3)TT(3) (5' GGGTTTGGGTGGGTTTGGG) is used, and the fluorescence of CV decreases with an increased concentration of K(+). The other is a fluorescence-increasing mode, in which Hum21 (5'-GGGTTAGGGTTAGGGTTAGGG) is used, and the fluorescence of CV increases with an increased concentration of K(+). Compared with some published K(+) detection methods, this method has some important characteristics, such as lower cost of the test, higher concentrations of Na(+) that can be tolerated, adjustable linear detection range and longer excitation and emission wavelengths. Preliminary results demonstrated that the method might be used in biological systems, for example in urine. PMID- 19559595 TI - Modular model-based design for heterologous bioproduction in bacteria. AB - We review the current status of expression of heterologous systems for bioenergy and bioproduction in bacteria using a model-based approach. As an aim for synthetic biology, it requires mathematical models of genetic modules that could be characterized independently of their context. This fastens the design of metabolic circuits using a combinatorial design approach, where given pathways could be optimized for maximal bioproduction, while being nontoxic for the chassis. We show how recent characterization of genetic parts, such as promoters, RBS or sRNAs could be used to fine-tune the expression of individual genes to achieve that goal. We also present lists of enzymes that are used for bioproduction, enlarging such set of biological parts. PMID- 19559597 TI - Calcium channel diversity: multiple roles of calcium channel subunits. AB - Until recently we held the simple view that voltage-gated calcium channels consisted of an alpha1 subunit, usually associated with auxiliary beta subunits and alpha(2)delta subunits and that skeletal muscle calcium channels were also associated with a gamma subunit. However, as discussed here, there is now evidence that the auxiliary subunits may also perform other roles unrelated to voltage-gated calcium entry. In the past students were taught the simplistic view that second messenger signaling to voltage-gated calcium channels involved mainly phosphorylation of L-type calcium channels, Ca(2+)-dependent inactivation via calmodulin, and direct G-protein-mediated inhibition of the neuronal N and P/Q channels. However, it is now clear that there are many other means of modulating calcium channel activity, including receptor-mediated internalization, proteolytic cleavage, phosphorylation of beta subunits, and interaction of calcium channels with other proteins, including enzymes masquerading as scaffold proteins. PMID- 19559596 TI - Junctional trafficking and epithelial morphogenesis. AB - Epithelial monolayers are major determinants of three-dimensional tissue organization and provide the structural foundation for the body plan and all of its component organs. Epithelial cells are connected by junctional complexes containing the cell adhesion molecule E-cadherin. Adherens junctions mediate stable cohesion between cells but must be actively reorganized to allow tissue remodeling during development. Recent studies demonstrate that junctional proteins are dynamically turned over at the cell surface, even in cells that do not appear to be moving. The redistribution of E-cadherin through spatially regulated endocytosis and exocytosis contributes to cell adhesion, cell polarity, and cell rearrangement. Here we describe recent progress in understanding the roles of the vesicle transport machinery in regulating cell adhesion and junctional dynamics during epithelial morphogenesis in vivo. PMID- 19559598 TI - Presynaptic signal transduction pathways that modulate synaptic transmission. AB - Presynaptic modulation is a crucial factor in the adaptive capacity of the nervous system. The coupling between incoming action potentials and neurotransmitter secretion is modulated by firstly, recent activity of the presynaptic axon that leads to the accumulation of residual calcium in the terminal and secondly, activation of presynaptic receptors by external signals. Despite the detailed description of these phenomena, the underlying mechanisms are still poorly understood. The nerve terminal contains many Ca(2+)-binding proteins that may contribute to the translation of residual Ca(2+)-increases to secretion modulation. We also found that >100 presynaptic proteins are phosphorylated and may contribute to the translation of presynaptic receptor activation to secretion modulation. However, which of these many candidates are the dominant regulators and how their activities integrate is largely unknown. Here, we review some of the recent insights into the complex interplay between presynaptic signal transduction components and propose blueprints of the major pathways. PMID- 19559600 TI - Fatty acid production from a highly CO2 tolerant alga, Chlorocuccum littorale, in the presence of inorganic carbon and nitrate. AB - Photoautotrophic fatty acid production of a highly CO(2)-tolerant green alga Chlorococcum littorale was investigated in the presence of inorganic carbon and nitrate at 295 K and a light intensity of 170 micromol-photon m(-2) s(-1). CO(2) concentration in the bubbling gas was adjusted by mixing pure gas components of CO(2) and N(2) to avoid photorespiration and beta-oxidation of fatty acids under O(2) atmosphere conditions. Fatty acid content was almost constant for the CO(2) concentrations ranging from 5% to 50% under nitrate-rich conditions corresponding to the logarithmic growth phase. After nitrate depletion, the content drastically increased with a decrease in CO(2) concentration. HCO(3)(-)/CO(2) ratio in the culture media was found to be a controlling factor for fatty acid production after the nitrate limitation phase. For a CO(2) concentration of 5%, the fatty acid content was ca. 34 wt.% at maximum, which is comparable with other land plant seed oils. PMID- 19559599 TI - Neutralizing ebolavirus: structural insights into the envelope glycoprotein and antibodies targeted against it. AB - The ebolavirus (EBOV) envelope glycoprotein (GP) is solely responsible for viral attachment to, fusion with, and entry of new host cells, and consequently is a major target of vaccine design efforts. Recently determined crystal structures of key antibodies in complex with their EBOV epitopes have provided insights into the molecular architecture of GP and defined likely hotspots for viral neutralization. In this review, we discuss the structural basis for antibody mediated neutralization of ebolavirus and its implications for novel therapeutic or vaccine strategies. PMID- 19559602 TI - Effects of spawn, supplement and phase II compost additions and time of re-casing second break compost on mushroom (Agaricus bisporus) yield and biological efficiency. AB - Three cropping experiments (0710, 0803 and 0805) were conducted to determine the effect of adding spawn, various levels of delayed release nutrient, and phase II compost to 2nd break mushroom compost (2BkC) on mushroom yield and biological efficiency (BE). We also investigated the effect of delaying time of re-casing non-supplemented and supplemented 2BkC on mushroom yields and BEs. The addition of 14.6% spawn to nutrient-supplemented 2BkC (w.w./d.w) increased yield by 11.1% over the control (no spawn) but did not affect BE. The addition of delayed release supplements to 2BkC increased maximum yields by 29-54%, depending on the treatment. Substitution of 15% phase II compost in 2BkC (15/85) did not significantly affect mushroom yields. However, use of 15% phase II compost in 2BkC increased the response of the mixture to delayed release supplement. Yield response to increasing levels of supplement was greater in the 15/85 mixture compared to 100% 2BkC. Yields also increased as time of re-casing was delayed up to 10 days. Mushroom yields increased approximately 2.1% for each day re-casing was delayed. Overall yields were generally higher from commercial 2BkC compared to 2BkC originating from the Penn State Mushroom Research Center (MRC) probably due to nitrogen (N) content of the 2BkC. Nitrogen content in commercial 2BkC (Crop 0805) was 3% while N content in 2BkC from Crops 0710 and 0803 was 2.2% and 2.1%, respectively. By optimizing supplement levels and adding 15% phase II compost to commercial 2BkC, or by delaying casing by 5-10 days, it was possible to obtain BEs that were equivalent to supplemented phase II compost. PMID- 19559601 TI - Knife mill operating factors effect on switchgrass particle size distributions. AB - Biomass particle size impacts handling, storage, conversion, and dust control systems. Switchgrass (Panicum virgatum L.) particle size distributions created by a knife mill were determined for integral classifying screen sizes from 12.7 to 50.8 mm, operating speeds from 250 to 500 rpm, and mass input rates from 2 to 11 kg/min. Particle distributions were classified with standardized sieves for forage analysis that included horizontal sieving motion with machined-aluminum sieves of thickness proportional to sieve opening dimensions. Then, a wide range of analytical descriptors were examined to mathematically represent the range of particle sizes in the distributions. Correlation coefficient of geometric mean length with knife mill screen size, feed rate, and speed were 0.872, 0.349, and 0.037, respectively. Hence, knife mill screen size largely determined particle size of switchgrass chop. Feed rate had an unexpected influence on particle size, though to a lesser degree than screen size. The Rosin-Rammler function fit the chopped switchgrass size distribution data with an R(2)>0.982. Mass relative span was greater than 1, which indicated a wide distribution of particle sizes. Uniformity coefficient was more than 4.0, which indicated a large assortment of particles and also represented a well-graded particle size distribution. Knife mill chopping of switchgrass produced 'strongly fine skewed mesokurtic' particles with 12.7-25.4 mm screens and 'fine skewed mesokurtic' particles with 50.8 mm screen. Results of this extensive analysis of particle sizes can be applied to selection of knife mill operating parameters to produce a particular size of switchgrass chop, and will serve as a guide for relations among the various analytic descriptors of biomass particle distributions. PMID- 19559603 TI - Biogas production and valorization by means of a two-step biological process. AB - The scope of this research work was to investigate biogas production and purification by a two-step bench-scale biological system, consisting of fed-batch pulse-feeding anaerobic digestion of mixed sludge, followed by methane enrichment of biogas by the use of the cyanobacterium Arthrospiraplatensis. The composition of biogas was nearly constant, and methane and carbon dioxide percentages ranged between 70.5-76.0% and 13.2-19.5%, respectively. Biogas yield reached a maximum value (about 0.4 m(3)(biogas)/kg COD(i)) at 50 days-retention time and then gradually decreased with a decrease in the retention time. Biogas CO(2) was then used as a carbon source for A. platensis cultivation either under batch or fed batch conditions. The mean cell productivity of fed-batch cultivation was about 15% higher than that observed during the last batch phase (0.035+/-0.006 g(DM)/L/d), likely due to the occurrence of some shading effect under batch growth conditions. The data of carbon dioxide removal from biogas revealed the existence of a linear relationship between the rates of A. platensis growth and carbon dioxide removal from biogas and allowed calculating carbon utilization efficiency for biomass production of almost 95%. PMID- 19559604 TI - Analysis of bacteria communities in an up-flow fixed-bed (UFB) bioreactor for treating sulfide in hydrocarbon wastewater. AB - An up-flow fixed-bed (UFB) bioreactor with patented functional polyurethane foam (FPUF) carriers was used to treat sulfide in hydrocarbon wastewater. Community compositions of autotrophic and heterotrophic bacteria were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). DGGE results showed that a relatively stable bacterial community composed of heterotrophic and autotrophic bacteria formed in the bioreactor by the end of experiment, which ensured 92-100% sulfide removal efficiencies. Furthermore, autotrophic genera of Thiobacillus and Thiomonas, as well as those of the heterotrophic genus of Acinetobacter survived and exhibited high sulfide oxidation activity under all three operational conditions. Different special genera were also observed under each operational condition, such as the halophilic genus of Nesterenkonia. In addition, a new genus of sulfide oxidation bacteria was found in the bioreactor, which had the ability to synthesize cytoplasm from organic compounds. These genera have wide applications for the treatment of sulfide in hydrocarbon wastewater. PMID- 19559605 TI - Landfill leachate treatment with microbial fuel cells; scale-up through plurality. AB - Three Microbial Fuel Cells (MFCs) were fluidically connected in series, with a single feed-line going into the 1st column through the 2nd column and finally as a single outflow coming from the 3rd column. Provision was also made for re circulation in a loop (the outflow from the 3rd column becoming the feed-line into the 1st column) in order to extend the hydraulic retention time (HRT) on treatment of landfill leachate. The effect of increasing the electrode surface area was also studied whilst the columns were (fluidically) connected in series. An increase in the electrode surface area from 360 to 1080 cm(2) increased the power output by 118% for C2, 151% for C3 and 264% for C1. COD and BOD(5) removal efficiencies also increased by 137% for C1, 279% for C2 and 182% for C3 and 63% for C1, 161% for C2 and 159% for C3, respectively. The system when configured into a loop was able to remove 79% of COD and 82% of BOD(5) after 4 days. These high levels of removal efficiency demonstrate the MFC system's ability to treat leachate with the added benefit of generating energy. PMID- 19559606 TI - Development of a pilot scale anaerobic digester for biogas production from cow manure and whey mix. AB - This paper presents results from anaerobic digestion of cow manure and whey mix. A pilot scale anaerobic digester, 128 l in volume, has been developed, to operate under batch and fed-batch conditions. The versatile and unique characteristics of the instrument allowed testing the methane production directly in the farm. The digester performance was evaluated with two calibration tests, the main for a period of 56 days. The study test was divided into three phases, one for each type of feeding operation (batch, fed-batch, batch). The initial phase of digestion resulted in 57 l-CH(4)/kg-VS, the second phase had a yield of 86.6 l CH(4)/kg-VS and the third one had a production of 67 l-CH(4)/kg-VS. The total methane yield was equal to 211.4 l-CH(4)/kg-VS. Using the obtained pilot plant results to a real scale diary production cycle, it was possible to evaluate an electricity production equal to 8.86 kwh per 1 t/d. The conducted tests did show that there is a good potential to the use of a cow manure and whey biomass mix for biogas production. PMID- 19559607 TI - An efficient synthesis and biological study of novel indolyl-1,3,4-oxadiazoles as potent anticancer agents. AB - A facile, convenient and high yielding synthesis of a series of novel 5-(3' indolyl)-2-(substituted)-1,3,4-oxadiazoles from readily available starting materials has been described. The key step of this protocol is oxidative cyclization of N-acylhydrazones 1 using [bis(trifluoroacetoxy)iodo]benzene under solvent-free condition. The 5-(3'-indolyl)-2-(substituted)-1,3,4-oxadiazoles were screened for their in vitro anticancer activity against various human cancer cell lines. Compounds 3c, 3d and 3j exhibited potent cytotoxicity (IC(50) approximately 1microM) and selectivity against human cancer cell lines. PMID- 19559608 TI - Functionalization of the A ring of pyridoacridine as a route toward greater structural diversity. Synthesis of an octacyclic analogue of eilatin. AB - In an effort to increase the structural diversity of pyrido[4,3,2-kl]acridines, compounds containing amino substituents on the A ring were synthesized. The key reactions involve regioselective electrophilic aromatic substitutions. The methodology was applied to the synthesis of the extended angular octacycle 8, which conjugates the physicochemical and spectroscopic properties of the pyridoacridine skeleton with the ability of [1,10]phenanthroline ring for metal complexation. The 9-aminopyridoacridine 4 displays significant cytostatic activities against two cancer cell lines, and may be considered as a new lead in the search of active derivatives. PMID- 19559610 TI - Positive sentinel lymph node biopsy predicts the number of metastatic axillary nodes of breast cancer. AB - It remains to be clarified whether a positive sentinel lymph node biopsy (SLNB) can predict the number of metastatic axillary nodes. This study examined a consecutive series of women with unilateral invasive breast cancer who underwent axillary lymph node dissection after an intra-operative positive SLNB. The numbers of positive and negative sentinel lymph nodes (SLNs) were analyzed for a likelihood of pN1a, pN2a, and pN3a diseases as per the UICC TNM classification. Of the 368 study patients, 165 (45%) had one positive SLN and one or more negative SLNs. This result represented the most common combination of positive and negative SLNs. It was also the most predictive indicator (93%) of pN1a disease and the least predictive indicator (7% or 0%) of pN2a or pN3a disease, respectively. The numbers of positive and negative SLNs can predict the number of metastatic axillary nodes in breast cancer patients. PMID- 19559609 TI - Peperomins as anti-inflammatory agents that inhibit the NF-kappaB signaling pathway. AB - The transcription factor nuclear factor kappaB (NF-kappaB) induces the expression of various inflammatory genes. In the common NF-kappaB signaling pathway, peperomin E and 2,6-didehydropeperomin B inhibited IkappaB degradation upon stimulation with TNF-alpha or interleukin-1. Consistent with these results, peperomin E and 2,6-didehydropeperomin B blocked the TNF-alpha-induced activation of IkappaB kinase, while they had no direct effect on the IkappaB kinase activity. Our present results clearly demonstrate that peperomins inhibit the NF kappaB signaling pathway by blocking IkappaB kinase activation. PMID- 19559611 TI - Localization of impalpable breast lesions: what are we aiming at? AB - The European published target for wire placement is 95% being within 10 mm of impalpable breast lesions. Other suggested targets may be more relevant. We assessed 500 procedures of which 411 could be measured. RESULTS: Fail 10 mm target: 3 (0.7%); Fail 5 mm target: 11 (2.7%); Fail 2 mm target: 21 (4.9%); within 2 mm: 11 (2.7%); and Fail to traverse lesion in one or more projection: 32 (7.8%). CONCLUSION: The European QA target has little supporting evidence and is easily met. Success in traversing lesions is a more measurable, achievable aim with clinical relevance. We suggest that this could be an improved target. PMID- 19559612 TI - EEG responses to visual landmarks in flying pigeons. AB - BACKGROUND: GPS analysis of flight trajectories of pigeons can reveal that topographic features influence their flight paths. Recording electrical brain activity that reflects attentional processing could indicate objects of interest that do not cause changes in the flight path. Therefore, we investigated whether crossing particular visual landmarks when homing from a familiar release site is associated with changes in EEG. RESULTS: Birds carried both data-loggers for recording GPS position and EEG during flight. First, we classified characteristic EEG frequencies of caged birds and found five main bands: A: 0-3, B: 3-12, C: 12 60, D: 60-130, and E: 130-200 Hz. We analyzed changes in these activity bands when pigeons were released over sea (a featureless environment) and over land. Passing over the coastline and other prominent landmarks produced a pattern of EEG alterations consisting of two phases: activation of EEG in the high-frequency bands (D and/or E), followed by activation of C. Overlaying the EEG activity with GPS tracks allowed us to identify topographical features of interest for the pigeons that were not recognizable by distinct changes of their flight path. CONCLUSIONS: We provide evidence that EEG analysis can identify landmarks and objects of interest during homing. Middle-frequency activity (C) reflects visual perception of prominent landmarks, whereas activation of higher frequencies (D and E) is linked with information processing at a higher level. Activation of E bands is likely to reflect an initial process of orientation and is not necessarily linked with processing of visual information. PMID- 19559613 TI - Thatcher effect in monkeys demonstrates conservation of face perception across primates. AB - Accurate recognition of individuals is a foundation of social cognition. The remarkable ability of humans to distinguish among thousands of similar faces depends on sensitivity to unique configurations of facial features, including subtle differences in the relative placement of the eyes and mouth. Determining whether similar perceptual processes underlie individual recognition in nonhuman primates is important for both the study of cognitive evolution and the appropriate use of primate models in social cognition research. In humans, some of the best evidence for a keen sensitivity to the configuration of features in faces comes from the "Thatcher effect." This effect shows that it is difficult to detect changes in the orientation of the eyes and mouth in an image of an inverted face, even though identical changes are unmistakable in an upright face. Here, we demonstrate for the first time that a nonhuman primate species also exhibits the Thatcher effect. This direct evidence of configural face perception in monkeys, collected under testing conditions that closely parallel those used with humans, indicates that perceptual mechanisms for individual recognition have been conserved through primate cognitive evolution. PMID- 19559614 TI - Dosage-dependent deregulation of an AGAMOUS-LIKE gene cluster contributes to interspecific incompatibility. AB - Postzygotic lethality of interspecies hybrids can result from differences in gene expression, copy number, or coding sequence and can be overcome by altering parental genome dosage. In crosses between Arabidopsis thaliana and A. arenosa, embryo arrest is associated with endosperm hyperproliferation and delayed development similar to paternal-excess interploidy crosses and polycomb repressive complex (PRC) mutants. Failure is accompanied by parent-specific loss of gene silencing including the dysregulation of three genes suppressed by PRC. Increasing the maternal genome dosage rescues seed development and gene silencing. A gene set upregulated in the failing seed transcriptome encoded putative AGAMOUS-LIKE MADS domain transcription factors (AGL) that were expressed in normal early endosperm and were shown to interact in a previous yeast 2-hybrid analysis. Suppression of these AGL's expression upon cellularization required PRC. Preceding seed failure, expression of the PRC member FIS2 decreased concomitant with overexpression of the AGL cluster. Inactivating two members, AGL62 and AGL90, attenuated the postzygotic barrier between A. thaliana and A. arenosa. We present a model where dosage-sensitive loss of PRC function results in a dysregulated AGL network, which is detrimental for early seed development. PMID- 19559615 TI - Centralization of the deuterostome nervous system predates chordates. AB - The origin of the chordate central nervous system (CNS) is unknown. One theory is that a CNS was present in the first bilaterian and that it gave rise to both the ventral cord of protostomes and the dorsal cord of deuterostomes. Another theory proposes that the chordate CNS arose by a dramatic process of dorsalization and internalization from a diffuse nerve net coextensive with the skin of the animal, such as enteropneust worms (Hemichordata, Ambulacraria) are supposed to have. We show here that juvenile and adult enteropneust worms in fact have a bona fide CNS, i.e., dense agglomerations of neurons associated with a neuropil, forming two cords, ventral and dorsal. The latter is internalized in the collar as a chordate-like neural tube. Contrary to previous assumptions, the greater part of the adult enteropneust skin is nonneural, although elements of the peripheral nervous system (PNS) are found there. We use molecular markers to show that several neuronal types are anatomically segregated in the CNS and PNS. These neuroanatomical features, whatever their homologies with the chordate CNS, imply that nervous system centralization predates the evolutionary separation of chordate and hemichordate lineages. PMID- 19559616 TI - RAN GTPase is a RASSF1A effector involved in controlling microtubule organization. AB - RASSF1A is a tumor suppressor gene that is inactivated by hypermethylation of its promoter region in most types of human cancers. The incidence of spontaneous or induced tumors is significantly higher in Rassf1a(-/-) mice than in wild-type mice, confirming the tumor suppressor function of RASSF1A. RASSF1A promotes apoptosis mainly through its interaction with the proapoptotic serine/threonine STE20-like kinases MST1 and 2. However, Rassf1a(-/-) mice do not show overt signs of deregulated apoptosis, suggesting that other RASSF1A effectors are also critical for tumor suppression. In a proteomics screen, we identified RAN GTPase, MST1 and 2 kinases, and alpha- and gamma-tubulin as RASSF1A-interacting proteins. We show that RASSF1A-induced microtubule hyperstability, a hallmark of RASSF1A expression, is RAN-GTP dependent. RASSF1A promotes the accumulation of the GTP bound form of RAN via the MST2-induced phosphorylation of RCC1. Depletion of RASSF1A results in mislocalization of RCC1 to the mitotic spindle and spindle poles, leading to mitotic spindle abnormalities and prometaphase block. A similar mitotic delay is also observed with MST2 depletion. These findings reveal a mechanism for how RASSF1A controls microtubule stability and for how its loss compromises the integrity of the mitotic spindle, leading to aneuploidy and tumorigenesis. PMID- 19559617 TI - Synergism of red and blue light in the control of Arabidopsis gene expression and development. AB - The synergism between red and blue light in the control of plant growth and development requires the coaction of the red light photoreceptor phytochrome B (phyB) and the blue light and UV-A receptor cryptochromes (cry). Here, we describe the mechanism of the coaction of these photoreceptors in controlling both development and physiology. In seedlings grown under red light, a transient supplement with blue light induced persistent changes in the transcriptome and growth patterns. Blue light enhanced the expression of the transcription factors LONG HYPOCOTYL 5 (HY5) and HOMOLOG OF HY5 (HYH) and of SUPPRESSOR OF PHYA 1 (SPA1) and SPA4. HY5 and HYH enhanced phyB signaling output beyond the duration of the blue light signal, and, contrary to their known role as repressors of phyA signaling, SPA1 and SPA4 also enhanced phyB signaling. These observations demonstrate that the mechanism of synergism involves the promotion by cry of positive regulators of phyB signaling. The persistence of the light-derived signal into the night commits the seedling to a morphogenetic and physiological program consistent with a photosynthetic lifestyle. PMID- 19559619 TI - Social cognition modulates the sensory coding of observed gaze direction. AB - Gaze direction is an important social signal in both human and nonhuman primates, providing information about conspecifics' attention, interests, and intentions. Single-unit recordings in macaques have revealed neurons selective for others' specific gaze direction. A parallel functional organization in the human brain is indicated by gaze-adaptation experiments, in which systematic distortions in gaze perception following prolonged exposure to static face images reveal dynamic interactions in local cortical circuitry. However, our understanding of the influence of high-level social cognition on these processes in monkeys and humans is still rudimentary. Here we show that the attribution of a mental state to another person determines the way in which the human brain codes observed gaze direction. Specifically, we convinced observers that prerecorded video sequences of an experimenter gazing left or right were a live video link to an adjacent room. The experimenter wore mirrored goggles that observers believed were either transparent such that the person could see, or opaque such that the person could not see. The effects of adaptation were enhanced under the former condition relative to the latter, indicating that high-level sociocognitive processes shape and modulate sensory coding of observed gaze direction. PMID- 19559618 TI - Allelic polymorphism within the TAS1R3 promoter is associated with human taste sensitivity to sucrose. AB - Human sweet taste perception is mediated by the heterodimeric G protein-coupled receptor encoded by the TAS1R2 and TAS1R3 genes. Variation in these genes has been characterized, but the functional consequences of such variation for sweet perception are unknown. We found that two C/T single-nucleotide polymorphisms (SNPs) located at positions -1572 (rs307355) and -1266 (rs35744813) upstream of the TAS1R3 coding sequence strongly correlate with human taste sensitivity to sucrose and explain 16% of population variability in perception. By using a luciferase reporter assay, we demonstrated that the T allele of each SNP results in reduced promoter activity in comparison to the C alleles, consistent with the phenotype observed in humans carrying T alleles. We also found that the distal region of the TAS1R3 promoter harbors a composite cis-acting element that has a strong silencing effect on promoter activity. We conclude that the rs307355 and rs35744813 SNPs affect gene transcription by altering the function of this regulatory element. A worldwide population survey reveals that the T alleles of rs307355 and rs35744813 occur at lowest frequencies in European populations. We propose that inherited differences in TAS1R3 transcription account for a substantial fraction of worldwide differences in human sweet taste perception. PMID- 19559621 TI - IF(1): setting the pace of the F(1)F(o)-ATP synthase. AB - When mitochondrial function is compromised and the mitochondrial membrane potential (Deltapsi(m)) falls below a threshold, the F(1)F(o)-ATP synthase can reverse, hydrolysing ATP to pump protons out of the mitochondrial matrix. Although this activity can deplete ATP and precipitate cell death, it is limited by the mitochondrial protein IF(1), an endogenous F(1)F(o)-ATPase inhibitor. IF(1), therefore, preserves ATP at the expense of Deltapsi(m). Despite a wealth of detailed knowledge on the biochemistry of the interaction of IF(1) and the F(1)F(o)-ATPase, little is known about its physiological activity. Emerging research suggests that IF(1) has a wider ranging impact on mitochondrial structure and function than previously thought. PMID- 19559620 TI - Mitochondrial DNA replication and repair: all a flap. AB - The mitochondrial genome is dwarfed by its neighbour in the nucleus, and, thus, it has been sensible for far more resources to be invested in the study of nuclear, rather than mitochondrial, DNA metabolism. Furthermore, few researchers have considered using mitochondrial DNA (mtDNA) as a model system for nuclear DNA metabolism. A quick look into the history of mtDNA provides ready answers as to why this was the case; however, recently mitochondria have been found to contain several nuclear replication and repair factors, so is there any potential to adopt the mitochondrion as a tool to unravel some of the intricacies of replication and repair in higher-order eukaryotes? Perhaps it is now time to invite the Cinderella genome to the ball. PMID- 19559622 TI - The tau of MARK: a polarized view of the cytoskeleton. AB - Microtubule-affinity regulating kinases (MARKs) were originally discovered by their ability to phosphorylate tau protein and related microtubule-associated proteins (MAPs), and thereby to regulate microtubule dynamics in neurons. Members of the MARK (also known as partition-defective [Par]-1 kinase) family were subsequently found to be highly conserved and to have key roles in cell processes such as determination of polarity, cell-cycle control, intracellular signal transduction, transport and cytoskeleton. This is important for neuronal differentiation, but is also prominent in neurodegenerative 'tauopathies' such as Alzheimer's disease. The identified functions of MARK/Par-1 are diverse and require accurate regulation. Recent discoveries including the x-ray structure of human MARKs contributed to an increased understanding of the mechanisms that control the kinase activity and, thus, the actin and microtubule cytoskeleton. PMID- 19559624 TI - Enaminones 10. Molecular modeling aspects of the 5-methylcyclohexenone derivatives. AB - This article expands upon our original submission to the Eddington, N. D.; Cox, D. S.; Khurana, M.; Salama, N. N.; Stables, J. P.; Harrison, S. J.; Negussie, A.; Taylor, R. S.; Tran, U. Q.; Moore, J. A.; Barrow, J. C.; Scott, K. R. Eur. J. Med. Chem.2003, 38, 49 on a series of twenty (20) compounds, all 5-methyl-3 [(substituted)-phenylamino]-cyclohex-2-enone derivatives. This article provides the reasons why the compounds are active/inactive. By use of computational methods, the reasons for activity/inactivity are explained. PMID- 19559623 TI - 'Click' D(1) receptor agonists with a 5-HT(1A) receptor pharmacophore producing D(2) receptor activity. AB - A series of new 1-aryl-3-benzazepine derivatives containing an arylpiperazinyl function as the N3 substituent were synthesized by combining a D(1) receptor agonistic pharmacophore and a 5-HT(1A) receptor pharmacophore through Click reaction. Interestingly, these compounds generally do not have good binding affinity at the D(1) receptor, but most compounds are potent at both D(2) and 5 HT(1A) receptors. Compound 8h, containing 1-m-tolyl-benzazepine scaffold and 2 methoxyphenylpiperazine core, displayed good affinity at all tested receptors, with K(i) values of 144, 80, and 133nM, for the D(1), D(2), and 5-HT(1A) receptors, respectively. Compound 13 with the triazole moiety formed differently from that in 8h showed the highest affinity at the D(2) receptor with K(i) value of 19nM. This compound also showed moderate affinity at the 5-HT(1A) (K(i), 105nM), and D(1) (K(i), 551nM) receptors. Functional assays indicated that both compounds 13 and 8h are antagonists at D(1) and D(2) receptors, whereas full agonistic activity at the 5-HT(1A) receptor was observed. In agreement with the binding affinity, compound 13 is a high efficacy D(2) antagonist and 5-HT(1A) agonist. PMID- 19559625 TI - A treatment algorithm for adult ameloblastomas according to the Pitie-Salpetriere Hospital experience. AB - During a 13-year period (from 1994 to 2007), in the Oral and Maxillofacial Surgery Department of the Pitie-Salpetriere Hospital, 116 new cases of adult ameloblastomas, were analyzed for treatment composed against radiographic presentation, size, histological type. Follow-up and recurrence were also analyzed. Treatment was surgical consisting of enucleations (82%), segmental mandibulectomy (8.3%) resections (24.7%) 85% of them underwent reconstruction. The follow-up was documented for 97%. More than two recurrences occurred in 21% of the patients after the first enucleation: 66% with a "follicular" histological diagnosis. Lenthly, a therapeutic algorithm is suggested for adult ameloblastomas that underlines the importance of the conservative enucleation treatment as far as possible. PMID- 19559626 TI - Exploring fluorescence and fragmentation of ions produced by electrospray ionization in ultrahigh vacuum. AB - Fluorescence spectroscopy and mass spectrometry have been extensively used for characterization of biomaterials, but usually separately. An instrument combining fluorescence spectroscopy and Fourier-transform ion cyclotron resonance mass spectrometry (FTICR-MS) has been developed to explore both fluorescence and mass spectrometric behavior of ions produced by electrospray ionization (ESI) in ultra high vacuum (<5 x 10(-9) mbar). Using rhodamine 6G (R6G) as a sample, the instrument was systematically characterized. Gas-phase fluorescence and mass spectral signal of the same ion population are detected immediately after each other. Effects of gas pressure, ion density, and excitation laser power on the fluorescence signal intensity and mass spectral fragmentation patterns are discussed. Characteristic times of ion photodissociation in ultra high vacuum were recorded for different irradiation powers. Photofragmentation patterns of rhodamine 6G ions in the Penning trap of an FTICR spectrometer obtained by photoinduced dissociation (PID) with visible light and sustained off-resonance irradiation collision-induced dissociation (SORI-CID) were compared. The lowest energy dissociation fragment of rhodamine 6G ions was identified by relating PID patterns of rhodamine 6G and rhodamine 575 dyes at various irradiation powers. The unique instrument provides a powerful platform for probing the intramolecular relaxation mechanisms of nonsolvated ions when interacting with light, which is of great fundamental interest for better understanding of their physical and chemical properties. PMID- 19559627 TI - Surface assisted laser desorption/ionization on two-layered amorphous silicon coated hybrid nanostructures. AB - Matrix-free laser desorption/ionization was studied on two-layered sample plates consisting of a substrate and a thin film coating. The effect of the substrate material was studied by depositing thin films of amorphous silicon on top of silicon, silica, polymeric photoresist SU-8, and an inorganic-organic hybrid. Des arg(9)-bradykinin signal intensity was used to evaluate the sample plates. Silica and hybrid substrates were found to give superior signals compared with silicon and SU-8 because of thermal insulation and compatibility with amorphous silicon deposition process. The effect of surface topography was studied by growing amorphous silicon on hybrid micro- and nanostructures, as well as planar hybrid. Compared with planar sample plates, micro- and nanostructures gave weaker and stronger signals, respectively. Different coating materials were tested by growing different thin film coatings on the same substrate. Good signals were obtained from titania and amorphous silicon coated sample plates, but not from alumina coated, silicon nitride coated, or uncoated sample plates. Overall, the strongest signals were obtained from oxygen plasma treated and amorphous silicon coated inorganic-organic hybrid, which was tested for peptide-, protein-, and drug molecule analysis. Peptides and drugs were analyzed with little interference at low masses, subfemtomole detection levels were achieved for des-arg(9) bradykinin, and the sample plates were also suitable for ionization of small proteins. PMID- 19559628 TI - Postdeployment hospitalizations among service members deployed in support of the operations in Iraq and Afghanistan. AB - PURPOSE: There is significant public and veteran concerns over the impact of military deployments to Iraq and Afghanistan on veterans' health. This study investigates morbidity among deployers by uniquely comparing after-deployment hospitalizations to before-deployment hospitalizations and hospitalizations among nondeployers. METHODS: To compare after-deployment with before-deployment rates of morbidity, we examined active-duty military personnel who deployed for the first time in support of the current conflicts in 2003 or 2004 and had at least 12 months of service before deployment. We also compared the after-deployment hospitalizations to hospitalizations of personnel serving on active duty from May 2002 through May 2004 without a deployment from September 2001 through August 2006. This historical prospective investigation utilized Cox's proportional hazards time-to-event modeling. Hospitalizations for any cause and hospitalizations based on 14 broad diagnostic categories were examined. RESULTS: After adjusting for demographic and occupational variables, the after-deployment risk for any-cause hospitalization was greater in comparison with before deployment (hazard ratio, 1.57; 95% CI, 1.48-1.66) but lower in comparison with nondeployers (HR, 0.95, 95% CI, 0.92-0.98). CONCLUSIONS: Active-duty service members have an increased risk of hospitalization after deployment compared with before deployment but a lower risk when compared with nondeployers. PMID- 19559629 TI - Infectious and thromboembolic complications of arthroscopic shoulder surgery. AB - HYPOTHESIS: This study investigates the rate of infectious and thromboembolic complications in shoulder arthroscopy and their association with pharmacologic prophylaxis. MATERIALS AND METHODS: On behalf of the Italian Society for Knee Surgery, Arthroscopy, Sport Traumatology, Cartilage and Orthopaedic Technologies (SIGASCOT), we asked the members to complete an on-line Web survey about their experiences and strategies of prophylaxis in shoulder arthroscopy. RESULTS: In the period 2005-2006, 9385 surgeries were performed. We report 15 infections and 6 DVTs. The overall rate of infections was 0.0016 (1.6/1000) and the rate of DVTs was 0.0006 (0.6/1000) CONCLUSION: The association between infection and antibiotic prophylaxis was significant (P=0.01); however, the risk of DVTs was not decreased with heparin prophylaxis. LEVEL OF EVIDENCE: Level 3. PMID- 19559630 TI - A systematic review of the psychometric properties of the Constant-Murley score. AB - HYPOTHESIS: The purpose of this study was to conduct a systematic review of the psychometric evidence relating to Constant-Murley score. MATERIALS AND METHODS: A search of 3 databases (Medline, CINAHL, and EMBASE) and a manual search yielded 35 relevant publications. Pairs of raters used structured tools to analyze these articles, through critical appraisal and data extraction. A descriptive synthesis of the psychometric evidence was then performed. RESULTS: Quality ratings of 23% of the studies reviewed reached a level of 75% or higher. Studies evaluating the content validity of the Constant-Murley score suggest that the description in the original publication is insufficient to accomplish standardization between centers and evaluators. Despite this limitation, the Constant-Murley score correlates strongly (>or= 0.70) with shoulder-specific questionnaires, reaches acceptable benchmarks (rho > 0.80) for its reliability coefficients, and is responsive (effect sizes and standardized response mean > 0.80) for detecting improvement after intervention in a variety of shoulder pathologies. DISCUSSION: This systematic review provides evidence to support the use of the Constant Murley score for specific clinical and research applications but underscores the need for greater standardization and precaution when interpreting scores. Methods to improve standardization and measurement precision are needed. Responsiveness has been shown to be excellent, but some properties still need be evaluated, particularly those related to the absolute errors of measurement and minimal clinically important difference. CONCLUSION: Given the widespread acceptance for usage of the Constant-Murley score in clinical studies and early indications that the measure is responsive, studies defining more rigid standardization of the tools/procedures are needed. LEVEL OF EVIDENCE: Level 1. PMID- 19559632 TI - Molecular heterogeneity of glucose-6-phosphate dehydrogenase deficiency among the tribals in Western India. PMID- 19559631 TI - Suppression of interleukin-33 bioactivity through proteolysis by apoptotic caspases. AB - Interleukin-33 (IL-33) is a member of the IL-1 family and is involved in polarization of T cells toward a T helper 2 (Th2) cell phenotype. IL-33 is thought to be activated via caspase-1-dependent proteolysis, similar to the proinflammatory cytokines IL-1 beta and IL-18, but this remains unproven. Here we showed that IL-33 was processed by caspases activated during apoptosis (caspase-3 and -7) but was not a physiological substrate for caspases associated with inflammation (caspase-1, -4, and -5). Furthermore, caspase-dependent processing of IL-33 was not required for ST2 receptor binding or ST2-dependent activation of the NF-kappaB transcription factor. Indeed, caspase-dependent proteolysis of IL 33 dramatically attenuated IL-33 bioactivity in vitro and in vivo. These data suggest that IL-33 does not require proteolysis for activation, but rather, that IL-33 bioactivity is diminished through caspase-dependent proteolysis within apoptotic cells. Thus, caspase-mediated proteolysis acts as a switch to dampen the proinflammatory properties of IL-33. PMID- 19559633 TI - Perisylvian polymicrogyria, infantile spasms and arthrogryposis: the severe end of the spectrum of congenital bilateral perisylvian polymicrogyria. AB - Congenital bilateral perisylvian polymicrogyria (CBPP) is the most frequent type of polymicrogyria in children. A 3-month-old male patient is described here with the combination of CBPP, infantile spasms and arthrogryposis. Only four patients have been reported earlier in the literature with this combination. Three of them had epilepsy. These patients represent the more severe phenotype of CBPP, characterized by early onset of symptoms, epilepsy, mental retardation, pseudobulbar palsy and arthrogryposis. PMID- 19559635 TI - Cutaneous carriage of Malassezia species in healthy and seborrhoeic Sphynx cats and a comparison to carriage in Devon Rex cats. AB - Cutaneous carriage of Malassezia species yeast was investigated in 32 Sphynx cats, and in 10 domestic shorthair (DSH) cats. Samples for mycological culture were taken using contact plates and swabs at seven sites in each cat (left and right axillae and groin, left ear, claw fold on left front paw and the interdigital palmar web of the left front paw). Malassezia species were isolated from 26/32 Sphynx cats (81%) and from 0/10 DSH control cats. In five cases Malassezia species yeasts were isolated at a single site, in the remaining 21 Sphynx cats at multiple sites. A total of 73 Malassezia species isolates were made, of which 68 were Malassezia pachydermatis and five were lipid-dependent Malassezia. Five out of the 32 Sphynx had greasy seborrhoea, and all seborrhoeic cats had M pachydermatis isolated from their skin, at multiple sites. None of the 32 Sphynx had Malassezia species isolated from the ears. The difference in population sizes between Sphynx and DSH cats was significant (Por= 3 neuropsychiatric symptoms present. The most often endosed symptoms were anxiety (62%), depression (36%), irritability (26%), as well as disinhibition and apathy (24% each). Twenty two patients (44%) had a score >or= 4 on at least one of the NPI items: again, most frequently anxiety (17 patients; 34%), depression (13 patients; 26%) and apathy (9 patients; 18%). Therefore, even among stable, long-term treated patients with WD approximately 70% experienced psychiatric symptoms. PMID- 19559639 TI - Selective hydrogenation by polymer-encapsulated platinum nanoparticles prepared by an easy single-step sonochemical synthesis. AB - Polypyrrole-encapsulated platinum nanoparticles (PPy/Pt-NPs) prepared by an easy single-step sonochemical synthesis were used as catalysts for the liquid phase hydrogenation of substituted alkenes in methanol or methanol/water mixtures. Polypyrrole (PPy) coatings on the nanoparticles were able to act as nanoscopic filters for substrates molecules, and consequently substrate selectivity could be controlled in the catalytic processes. PMID- 19559641 TI - Spinocerebellar ataxia 8: variable phenotype and unique pathogenesis. AB - Spinocerebellar ataxia 8 (SCA8), a triplet repeat expansion disorder, is genetically distinct from the other inherited ataxias, but its unusually variable phenotype can make its diagnosis difficult. In this review we describe 3 new cases of genetically verified SCA8 to highlight the broad clinical spectrum of symptoms observed with this disorder and to draw attention to the features of myoclonus and migraine headaches, which in the context of cerebellar ataxia warrants the clinician to consider SCA8 as a potential diagnosis. We also address the controversy surrounding the genetic testing approach for diagnosing SCA8. Finally, we evaluate the evidence that SCA8 may affect calcium channel function and that the presentation of episodic ataxia and migraines suggests a clinical and pathogenic overlap of SCA8 with the channelopathies. PMID- 19559642 TI - Arabidopsis lateral root development: an emerging story. AB - Lateral root formation is a major determinant of root systems architecture. The degree of root branching impacts the efficiency of water uptake, acquisition of nutrients and anchorage by plants. Understanding the regulation of lateral root development is therefore of vital agronomic importance. The molecular and cellular basis of lateral root formation has been most extensively studied in the plant model Arabidopsis thaliana (Arabidopsis). Significant progress has recently been made in identifying many new Arabidopsis genes that regulate lateral root initiation, patterning and emergence processes. We review how these studies have revealed that the plant hormone auxin represents a common signal that integrates these distinct yet interconnected developmental processes. PMID- 19559644 TI - The free-energy principle: a rough guide to the brain? AB - This article reviews a free-energy formulation that advances Helmholtz's agenda to find principles of brain function based on conservation laws and neuronal energy. It rests on advances in statistical physics, theoretical biology and machine learning to explain a remarkable range of facts about brain structure and function. We could have just scratched the surface of what this formulation offers; for example, it is becoming clear that the Bayesian brain is just one facet of the free-energy principle and that perception is an inevitable consequence of active exchange with the environment. Furthermore, one can see easily how constructs like memory, attention, value, reinforcement and salience might disclose their simple relationships within this framework. PMID- 19559643 TI - Linking development to defense: auxin in plant-pathogen interactions. AB - Although the plant growth hormone auxin has long been recognized as a regulator of plant defense, the molecular mechanisms involved are still largely unknown. Recent studies reviewed here reveal new insights into the role of auxin in plant defense. Similar to the signaling pathways of the defense-associated plant hormones salicylic acid (SA) and jasmonic acid (JA), auxin signaling differentially affects resistance to separate pathogen groups. Recent evidence suggests that the auxin and SA pathways act in a mutually antagonistic manner during plant defense, whereas auxin and JA signaling share many commonalities. Auxin also affects disease outcomes indirectly through effects on development. Here, we discuss the multiple ways in which auxin regulation of plant growth and development might be intimately linked to plant defense. PMID- 19559645 TI - Screening for polyomavirus associated nephropathy in renal transplantation with blood viral load measurement. AB - BACKGROUND: Polyomavirus associated nephropathy (PVAN) is an important cause of graft failure in the renal transplant population. It has been shown that viremia precedes PVAN, suggesting that measurement of blood viral load could be used for PVAN screening. OBJECTIVES: To verify the utility of BK virus (BKV) blood viral load measurement for PVAN screening in the renal transplant population, establish a threshold value, and determine the sensitivity and specificity of the test. STUDY DESIGN: We developed a real-time PCR assay for BKV blood viral load measurement and included this assay in the PVAN screening protocol of the renal transplant recipients of our institution. We report results for 60 patients who had a blood viral load measurement concomitantly with an allograft biopsy with immunohistochemistry for polyomavirus. RESULTS: 14 patients were found to have a PVAN on allograft biopsy together with a viral load above 3.0x10(3)copies/ml. None of the patients with a viral load under 3.0x10(3)copies/ml had a PVAN on allograft biopsy. The area under the receiver operating characteristic (ROC) curve was 0.95 (95% CI: 0.91-1.00) and using a threshold value of 3.0x10(3)copies/ml yielded a sensitivity of 100% (95% CI: 76.8-100%) and a specificity of 89.6% (95% CI: 77.3-96.5%) for PVAN screening. CONCLUSIONS: BKV blood viral load measurement is sensitive and specific for PVAN screening when a threshold value is precisely determined. PMID- 19559646 TI - Automated-parameterization of the motor evoked potential and cortical silent period induced by transcranial magnetic stimulation. AB - OBJECTIVE: To standardize the characterization of motor evoked potential (MEP) and cortical silent period (CSP) recordings elicited with transcranial magnetic stimulation (TMS). METHODS: A computer-based, automated-parameterization program (APP) was developed and tested which provides a comprehensive set of electromyography (EMG) magnitude and temporal measures. The APP was tested using MEP, CSP, and isolated CSP (iCSP) TMS stimulus-response data from a healthy adult population (N=13). RESULTS: The APP had the highest internal reliability (Cronbach's alpha=.98) for CSP offset time compared with two prominent automated methods. The immediate post-CSP EMG recovery level was 49% higher than the pre TMS EMG level. MEP size (peak amplitude, mean amplitude, peak-to-peak amplitude, and area) correlated higher with effective E-field (E(eff)) than other intensity measures (r approximately 0.5 vs. r approximately 0.3) suggesting that E(eff) is better suited for standardizing MEP stimulus-response relationships. CONCLUSIONS: The APP successfully characterized individual and mean epochs containing MEP, CSP, and iCSP responses. The APP provided common signal and temporal measures consistent with previous studies and novel additional parameters. SIGNIFICANCE: With the use of the APP modeling method and the E(eff), a standard approach for the analysis and reporting of MEP-CSP complex and iCSP measurements is achievable. PMID- 19559647 TI - The nature of individual differences in EEG parameters during homogeneous visual field in 5- and 6-year-old twins: a pilot study. AB - OBJECTIVE: To assess the genetic and environmental influences on the inter individual variability of spectral EEG parameters under two conditions of reduced visual input in preschoolers. METHODS: EEG was recorded in 21 monozygotic and 20 same-sex dizygotic twin pairs aged 5-6 years under the Closed eyes and Darkness conditions. The contributions of genetic and environmental influences to individual EEG parameters were estimated using structural equation modeling. RESULTS: The alpha rhythm parameters - both spectral amplitude and frequency - were influenced mainly by genetic factors. Theta spectral amplitude variance was presumably accounted for by both genetic and shared environmental factors. The difference in the nature of population variance between EEG alpha and theta band spectral amplitudes was reproduced for two identical functional loads: Closed eyes and Darkness. CONCLUSIONS: The majority of EEG spectral parameters in preschool children are determined by genetic factors. Tentative finding on systematic environmental influences on theta spectral amplitude under the condition of reduced visual input may point to the role of experience in the individual differences in the functioning of theta response system. SIGNIFICANCE: The findings contribute to the scanty knowledge about the heritability of EEG rhythms in preschool children. PMID- 19559649 TI - The role of monitoring mosquito infection in the Global Programme to Eliminate Lymphatic Filariasis. AB - In addition to monitoring infection in the human host, there is also a need to assess larval infection in the vector mosquito population to evaluate the success of interventions for eliminating lymphatic filariasis transmission from endemic communities. Here, we review the current status of the available tools for quantifying vector infection and existing knowledge and evidence regarding potential infection thresholds for determining transmission interruption, to assess the potential for using vector infection monitoring as a tool for evaluating the success of filariasis treatment programmes. PMID- 19559648 TI - Gene copy number and malaria biology. AB - Alteration in gene copy number provides a simple way to change expression levels and alter phenotype. This was fully appreciated by bacteriologists more than 25 years ago, but the extent and implications of copy number polymorphism (CNP) have only recently become apparent in other organisms. New methods demonstrate the ubiquity of CNPs in eukaryotes and their medical importance in humans. CNP is also widespread in the Plasmodium falciparum genome and has an important and underappreciated role in determining phenotype. In this review, we summarize the distribution of CNP, its evolutionary dynamics within populations, its functional importance and its mode of evolution. PMID- 19559650 TI - Development and evaluation of a novel multiple-antigen ELISA for serodiagnosis of tuberculosis. AB - In this study, we describe the development and evaluation of a novel multiple antigen ELISA for rapid diagnosis and screening of active tuberculosis (TB). The humoral immune responses of 136 active TB patients and 57 healthy subjects against antigens Rv3425, 38kDa and lipoarabinomannan (LAM) from Mycobacterium tuberculosis H37Rv were examined by ELISA. Three essential results were obtained. (i) Rv3425 antigen is a potential candidate for serodiagnosis of active TB. Of 136 active TB patients, Rv3425 antigen provided a sensitivity of 31.6%, lower than that of LAM antigen, but higher than that of 38kDa antigen, with an overall specificity of 100%. (ii) For 62 smear-negative pulmonary TB patients and 15 extra-pulmonary TB patients, the multiple-antigen test provided a sensitivity of 43.5% and 26.7%, respectively, representing an improvement over acid-fast bacilli (AFB) smear-based diagnosis. (iii) Compared with the single-antigen ELISA and the two available commercial kits, the multiple-antigen test offered the highest accuracy (71.0%). In conclusion, the multiple-antigen ELSIA test based on Rv3425, 38kDa, and LAM antigens is a potentially useful tool for the serodiagnosis and screening of active TB. Combinations of Rv3425 with other mycobacterial antigens may also be worthy of further investigation. PMID- 19559651 TI - Liquefaction and cavity formation in pulmonary TB: a simple method in rabbit skin to test inhibitors. AB - To control tuberculosis in the world today an additional approach would be most welcomed. Preventing (or reducing) pulmonary cavity formation is one such approach that has been almost completely neglected. Pulmonary cavity formation and the extracellular growth of tubercle bacilli in cavities cause bronchial spread of the disease in adult patients and spread of the bacillus to the environment where they infect other people. Therefore, cavity formation perpetuates tuberculosis in mankind. If no cavities form, the patient is much less infectious. Also, cavity formation often allows the tubercle bacillus to multiply (extracellularly) to tremendous numbers. Therefore, in humans almost all multidrug-resistant tubercle bacilli develop in cavities. This communication reviews the literature on liquefaction and cavity formation, and lists some of the responsible hydrolytic enzymes. It also describes a simple method to identify inhibitory pharmaceuticals, i.e., to observe their effect on the liquefaction and ulceration of skin lesions produced in rabbits by ascending concentrations of live or dead tubercle bacilli. PMID- 19559652 TI - 26 years of LDL--apheresis: a review of experience. AB - Since 1981, when LDL-Apheresis was introduced into the clinical routine at the University of Cologne as the first and so far only Apoprotein B specific LDL cholesterol elimination technique, considerable experience has since then accumulated and has changed not only the operational technique but also extended the indications, the optimation of the target values, the introduction of supportive cholesterol lowering drug therapy, considerations of the potential pleiotropic mechanisms and the introduction of a quality control supported electronic data processing. Mild to moderate side effects range between 3% and 4.5%, whereas serious undesired reactions did not occur within 26 years with more than 80,000 treatments performed at Cologne and considerably more world wide. As cholesterol can nowadays be widely eliminated in patients with familial hypercholesterolaemia (FH), the focus of consideration should be more directed to the treatment of additional risk factors. Thus, centres of competence, providing for more than the ability to technically reduce cholesterol may be desirable. Whereas numerous diagnostic procedures exist to demonstrate the value of cholesterol lowering therapies, the prolongation of survival as demonstrated in 7 homozygous and 29 heterozygous FH patients and in 5 patients with end stage disease appears to be the most convincing evidence for the value of LDL Apheresis. Due to the repetitive cycling and re-use LDL-Apheresis is furthermore not only the most efficient but also the most economic approach to extracorporeal LDL-elimination therapy. PMID- 19559653 TI - First-in-man clinical results of the treatment of patients with graft versus host disease with human ex vivo expanded CD4+CD25+CD127- T regulatory cells. AB - Here, we describe a procedure and first-in-man clinical effects of adoptive transfer of ex vivo expanded CD4+CD25+CD127- T regulatory cells (Tregs) in the treatment of graft versus host disease (GvHD). The cells were sorted from buffy coats taken from two family donors, expanded ex vivo and transferred to respective recipients who suffered from either acute or chronic GvHD. The therapy allowed for significant alleviation of the symptoms and reduction of pharmacologic immunosuppression in the case of chronic GvHD, while in the case of grade IV acute GvHD it only transiently improved the condition, for the longest time within all immunosuppressants used nonetheless. PMID- 19559655 TI - The lived experience of initial symptoms of and factors triggering epileptic seizures. AB - The aim of this study was to document the self-perception of initial symptoms of and factors triggering epileptic seizures in a sample of people with epilepsy (PWE) and their carers. Among 600 participants, questionnaires were returned by 309 (51.5%), of whom 72.8% were PWE and 27.2% were carers and others. Experiencing at least one symptom prior to a seizure was reported by 86.9% of PWE and 74% of carers. The most common symptoms were a funny feeling, confusion, and anxiety. Experiencing one trigger that resulted in a seizure was reported by 89.8% of PWE and 85.5% of carers. The most common triggers were tiredness, stress, and sleep deprivation. Among PWE and their carers, 63.6% and 51.3%, respectively, indicated that they can tell when a seizure is about to occur, and 26.7% and 15.4%, respectively, indicated that they felt they could stop a seizure. The most common techniques were resting, medication, and relaxation. PMID- 19559656 TI - Internal qigong for pain conditions: a systematic review. AB - The objective of this systematic review was to assess the evidence for the effectiveness of internal qigong as a treatment option for pain conditions. Nineteen databases were searched through to February 2009. Controlled clinical trials testing internal qigong in patients with pain of any origin assessing clinical outcome measures were considered. Trials using any type of internal qigong and control intervention were included. The selection of studies, data extraction, and validation were performed independently by 2 reviewers. Four randomized clinical trials (RCTs) and 3 controlled clinical trials met all inclusion criteria. One RCT suggested no significant difference for low back pain compared with electromyographic biofeedback. Two RCTs failed to show effects of internal qigong in neck pain compared with exercise therapy and waiting list control. One RCT suggested that qigong is inferior to aerobic exercise in patients with fibromyalgia. There are few RCTs testing the effectiveness of internal qigong in the management of pain conditions. Collectively, the existing trial evidence is not convincing enough to suggest that internal qigong is an effective modality for pain management. PERSPECTIVE: This review of controlled clinical trials focused on the effects of internal qigong, a self-directed energy healing intervention involving movement and meditation. Collectively, the existing trial evidence is not convincing enough to suggest that internal qigong is an effective modality for pain management. Future studies should be of high quality with particular emphasis on designing an adequate control intervention. PMID- 19559654 TI - Interferon-beta treatment in multiple sclerosis attenuates inflammatory gene expression through inducible activity of the phosphatase SHP-1. AB - Interferon-beta is a current treatment for multiple sclerosis (MS). Interferon beta is thought to exert its therapeutic effects on MS by down-modulating the immune response by multiple potential pathways. Here, we document that treatment of MS patients with interferon beta-1a (Rebif) results in a significant increase in the levels and function of the protein tyrosine phosphatase SHP-1 in PBMCs. SHP-1 is a crucial negative regulator of cytokine signaling, inflammatory gene expression, and CNS demyelination as evidenced in mice deficient in SHP-1. In order to examine the functional significance of SHP-1 induction in MS PBMCs, we analyzed the activity of proinflammatory signaling molecules STAT1, STAT6, and NF kappaB, which are known SHP-1 targets. Interferon-beta treatment in vivo resulted in decreased NF-kappaB and STAT6 activation and increased STAT1 activation. Further analysis in vitro showed that cultured PBMCs of MS patients and normal subjects had a significant SHP-1 induction following interferon-beta treatment that correlated with decreased NF-kappaB and STAT6 activation. Most importantly, experimental depletion of SHP-1 in cultured PBMCs abolished the anti-inflammatory effects of interferon-beta treatment, indicating that SHP-1 is a predominant mediator of interferon-beta activity. In conclusion, interferon-beta treatment upregulates SHP-1 expression resulting in decreased transcription factor activation and inflammatory gene expression important in MS pathogenesis. PMID- 19559657 TI - Adenosine modulates alpha2-adrenergic receptors through a phospholipase C pathway in brainstem cell culture of rats. AB - Adenosine acts in the nucleus tractus solitarii (NTS), one of the main brain sites related to cardiovascular control. In the present study we show that A(1) adenosine receptor (A(1R)) activation promotes an increase on alpha(2) adrenoceptor (Alpha(2R)) binding in brainstem cell culture from newborn rats. We investigated the intracellular cascade involved in such modulatory process using different intracellular signaling molecule inhibitors as well as calcium chelators. Phospholipase C, protein kinase Ca(2+)-dependent, IP(3) receptor and intracellular calcium were shown to participate in A(1R)/Alpha(2R) interaction. In conclusion, this result might be important to understand the role of adenosine within the NTS regarding autonomic cardiovascular control. PMID- 19559658 TI - Inhaled medication and inhalation devices for lung disease in patients with cystic fibrosis: A European consensus. AB - In cystic fibrosis inhalation of drugs for the treatment of CF related lung disease has been proven to be highly effective. Consequently, an increasing number of drugs and devices have been developed for CF lung disease or are currently under development. In this European consensus document we review the current status of inhaled medication in CF, including the mechanisms of action of the various drugs, their modes of administration and indications, their effects on lung function, exacerbation rates, survival and quality of life, as well as side effects. Specifically we address antibiotics, mucolytics/mucous mobilizers, anti-inflammatory drugs, bronchodilators and combinations of solutions. Additionally, we review the current knowledge on devices for inhalation therapy with regard to optimal particle sizes and characteristics of wet nebulisers, dry powder and metered dose inhalers. Finally, we address the subject of testing new devices before market introduction. PMID- 19559659 TI - Fully automated method for simultaneous determination of total cysteine, cysteinylglycine, glutathione and homocysteine in plasma by HPLC with UV absorbance detection. AB - A fully automated HPLC method for the simultaneous determination of total thiols in plasma samples has been developed. The method involves reductive conversion of disulfides to their reduced counterparts with the use of tris(2 carboxyethyl)phosphine. After reduction the newly formed sulfhydryl groups are reacted with 2-chloro-1-methylquinolinium tetrafluoroborate to form 2-S quinolinium derivatives followed by deproteinization by dialysis. The reaction products are separated by reversed-phase HPLC, detected and quantified by UV absorbance detection at 355nm. The recommended HPLC procedure enables measurement of four main plasma aminothiols cysteine, cysteinylglycine, glutathione, and homocysteine with low imprecision (mean relative standard deviations within calibration range, 3.47%, 5.34%, 4.25% and 3.26%, respectively) and good sensitivity. Accuracy, expressed as the mean measured amount as percentage of added amount, was within 97.5-103.0%, 98.3-102.5%, 96.3-99.5% and 97.1-99.1%, respectively. The lower limit of quantification for all thiols was 0.5microM. The whole unattended instrument acquisition time amounts 13min. PMID- 19559661 TI - Oncoplastic breast surgery: a review and systematic approach. AB - Oncoplastic breast surgery (OBS) is relatively new, but has made rapid progress from its tentative steps of infancy in the 1990s. The recent Milanese Consensus Conference on Breast Conservation concluded that, firstly, oncoplastic techniques are warranted to allow wide excision and clear margins without compromising cosmesis. Secondly, such surgery is ideally performed at the same time as oncological excision. Whilst technically more challenging than standard breast conserving therapy (BCT), OBS is well proven, if not yet widely practised, both oncologically and aesthetically and a review of the available techniques is perhaps timely. The roots of breast conserving therapy can be traced to the 1930s, actually due to advances made in radiotherapy, and the last 20 years have seen it become firmly established. This review aims to summarise the key historical developments and latest innovations in OBS. Not only are our patients, who expect not only safe cancer treatment but a satisfactory aesthetic outcome, increasingly informed and demanding, but longer follow up has stimulated surgeons to improve outcomes. In many cases, particularly with ptosis and macromastia, the cancer can be treated, usually with wider excision margins, simultaneously improving the aesthetic appearance. Present at the birth of OBS, the Institut Curie has continued to introduce innovative techniques over the last two decades and a systematic approach, comprising nine basic techniques, has evolved to allow high quality treatment of any and all breast cancers suitable for OBS. PMID- 19559662 TI - Guidelines for reconstruction of the eyelids and canthal regions. AB - A large number of reports are available on the options for reconstructing specific defects of the eyelids or (peri) orbital area, which may complicate decision making in choosing the most optimal technique for a particular defect. Based on more than 40 years' experience in reconstruction of eyelids and periorbital defects, combined with an extensive literature review, general principles on reconstruction are presented and illustrated. Surgical techniques are outlined with respect to anatomical layer, depth, size and location of the defect. Adherence to specific principles for eyelid and canthal reconstruction will lead to predictable, stable and functionally good results. PMID- 19559663 TI - Insufficient pain relief after surgical neuroma treatment: Prognostic factors and central sensitisation. AB - BACKGROUND: Treatment of patients with neuromatous pain is difficult. Numerous treatment methods have been described, but none has been completely effective in providing sufficient pain relief. Patient-specific prognostic factors, predicting pain after surgical neuroma treatment, can help clinicians in the process of patient treatment and care. METHODS: A computerised bibliographical database (PubMed Medline) was searched for articles concerning prognostic factors predicting the outcome of surgical neuroma treatment, and all the reference lists were checked. RESULTS: Evidence for predicting the outcome was found for neuromas of the radial sensory branch and digital nerves, discrete nerve syndrome, workers compensation, employment status, litigation involvement, duration of pain and number of previous operations. Psychosocial problems are often found in neuroma patients. In chronic neuropathic pain patients, changes in the central nervous system at the level of spinal cord and in the somatosensory cortex can be found. CONCLUSIONS: Neuromas of the radial sensory branch and digital nerves, discrete nerve syndrome, workers' compensation, employment status, litigation involvement, duration of pain and number of previous operations appear to predict the amount of pain after neuroma surgery. However, in a minority of patients, a bad outcome cannot be explained by these factors; in these patients, central sensitisation and psychosocial factors may play a role in maintaining pain. Research focussing on prognostic factors and the central changes induced by painful peripheral injury can lead to new and improved clinical treatment algorithms for the relief and prevention of chronic neuropathic pain. PMID- 19559660 TI - Autophagy in tumour suppression and promotion. AB - Autophagy, a well-described cellular mechanism for lysosomal degradation of cytoplasmic content, has emerged as a tumour suppression pathway. Recent evidence indicates that the tumour suppressor function of autophagy is mediated by scavenging of damaged oxidative organelles, thereby preventing accumulation of toxic oxygen radicals that would cause genome instability. Paradoxically, however, in some cases autophagy can also promote the survival of cancer cells once tumours have developed. This is attributed to the ability of autophagy to promote cell survival under conditions of poor nutrient supply, as often faced by solid tumours and metastasising cancer cells. In addition, autophagy is frequently upregulated in tumours as a response to therapy and may protect tumours against therapy-induced apoptosis. In this review we discuss the mechanisms that link autophagy to tumour suppression and promotion and provide examples of the dual functions of autophagy in cancer. PMID- 19559664 TI - Upper extremity kinematics analysis in obstetrical brachial plexus palsy. AB - INTRODUCTION: Several recent studies demonstrate that upper extremities kinematics analysis is in increasing use to assist clinical practice. We describe an upper limb kinematics analysis protocol that was first applied to a group of healthy children (to obtain normative data), and subsequently, to a child presenting with obstetrical brachial plexus palsy (OBPP) before and after surgical treatment. MATERIALS AND METHODS: The protocol is based on two very simple tasks. Reflective markers are placed on the studied segments, and optoelectronic cameras three-dimensionally record the position of the markers during the course of movement. The data, collected by a Vicon system (Oxford Metrics Ltd., Oxford, UK), are analyzed by a dedicated software; this software provides coefficient of multiple correlation (CMC) for the comparison of different kinematics curves and motion amplitudes. A CMC above 0.95 was considered to be excellent, between 0.85 and 0.95 was good, and below 0.85 was poor. Twelve healthy children, average age 9.7 years (from 7 to 14 years), were analyzed. A 7-year-old patient presenting left OBPP was similarly analyzed, pre- and postoperatively, after a lateral rotation osteotomy of the humerus. RESULTS: The analysis of the 12 healthy children established a kinematics corridor for each task and each angle considered. Analysis of the pathological patient revealed kinematics anomalies during movement which went undetected at simple clinical examination. CMC analysis after treatment showed improvement of all movements around the shoulder, going from "poor" preoperatively to "excellent" postoperatively. Amplitudes analysis similarly demonstrated postoperative improvement, which increased from 28 to 67% according to the rotations considered, around the shoulder and elbow. The interest in these results should be confirmed by studies in a larger number of patients. DISCUSSION: Upper extremity kinematics analysis is increasingly utilized in current clinical practice. Although many problems occur because of the non-cyclical and non automatic nature of movement, review of the literature and our preliminary results show that reproducibility is satisfactory. Interest in our work arises from helping develop a preoperative evaluation tool (providing a more global view of abnormalities) as well as a postoperative assessment one (for the quantification of movement gains obtained by surgery after humeral osteotomy). LEVEL OF EVIDENCE: Level IV. Diagnostic retrospective study. PMID- 19559665 TI - Design and validation of a high-throughput assay to detect codon 146 polymorphisms in the caprine prion protein gene. AB - In sheep, scrapie susceptibility is so strongly associated with single nucleotide polymorphisms (SNPs) in the gene encoding the prion protein (PrP) that this linkage constitutes the basis for selective breeding strategies directed toward controlling the disease. For goats, in contrast, the association between scrapie susceptibility/resistance and variations in the PrP gene is far weaker, with only a few identified SNPs showing an influence on scrapie susceptibility. A recent survey of PrP genotypes in Cypriot goats, however, revealed the existence of a robust association between polymorphisms at codon 146 of the caprine PrP gene and resistance/susceptibility to natural scrapie. Here we describe here a high throughput assay, based on homogeneous MassExtend technology coupled with mass spectrometry, for genotyping codon 146 of the caprine PrP gene. Our results demonstrate that this assay exhibits high accuracy, reproducibility, and repeatability, thereby making it suitable for large-scale SNP genotyping, as required for scrapie surveillance programs. PMID- 19559667 TI - Development of effective isolation method of ES cells for analysis of differentiation. AB - Neuroectoderm development is a milestone of vertebrate neurogenesis. However, the molecular mechanism underlying the differentiation of neuroectoderm is still unclear, especially in mammals. ES cells co-cultured with PA6 cells can differentiate to neuroectoderm by the stromal cell-derived inducing activity method (SDIA method), but contamination of PA6 cells is an obstacle to the analysis of molecular mechanisms of differentiation. Here we describe a novel method by which differentiated ES cells are easily isolated from PA6 cells. We attempted to induce the differentiation of ES cells using paraformaldehyde-fixed PA6 cells. RT-PCR and DNA microarray analysis revealed that the background noise derived from contaminated PA6 cells disappeared when fixed PA6 cells were used. Furthermore, genes up-regulated during the differentiation of ES cells were expressed in a developing mouse embryo. Thus, our newly developed method will be very useful for identifying novel genes associated with mouse neuroectoderm development in vitro and in vivo. PMID- 19559666 TI - Anti-inflammatory peptides grab on to the whiskers of atherogenic oxidized lipids. AB - The peptide 4F is known to have potent anti-atherogenic activity. 4F is an 18 residue peptide that has a sequence capable of forming a class A amphipathic helix. Several other class A amphipathic helical, 18 residue peptides with the same polar face but with increasing Phe residues on the nonpolar face have been synthesized with varying degrees of biological activity. In this work we compared the properties of the original 2F peptide, modeled on the consensus sequence of the amphipathic helical segments of the apolipoprotein A-I with the peptide 4F that has two Leu residues replaced with Phe. We demonstrate that the more biologically active 4F peptide has the greatest affinity for binding to several molecular species of oxidized lipids. Lipoprotein particles can be formed by solubilizing 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC) with peptides. These solubilized lipoprotein particles extract oxidized lipid from liposomes of POPC containing 5 mol% of oxidized lipid. The peptides with the strongest anti atherogenic activity interact most strongly with the oxidized lipid. The results show that there is a correlation between the biological potency of these peptides and their ability to interact with certain specific cytotoxic lipids, suggesting that this interaction may contribute favourably to their biological properties. PMID- 19559668 TI - Antigen-specific IL-23/17 pathway activation by murine semi-mature DC-like cells. AB - We analyzed the phenotype and function of bone marrow-derived dendritic cells (DCs) induced in vitro without using any serum during the late stage of cultivation. These 'serum-free' DCs (SF-DCs) possessed the ability to induce T cell proliferation as well as antibody responses, indicating that they were functional DCs. Surprisingly, the SF-DCs akin to semi-mature DCs in terms of both phenotypic and functional characteristics. The SF-DCs did not produce IL-12 but produced large amounts of IL-23 following lipopolysaccharide stimulation. The antigen-specific production of IL-17 by CD4(+) T cells co-cultured with OVA loaded SF-DCs was significantly higher than that with OVA-loaded conventional DCs. These results suggest that SF-DCs tend to produce IL-23 and can consequently induce the IL-17 producing CD4(+) T cells. The semi-mature DC-like cells reported here will be useful vehicles for DC immunization and might contribute to studies on the possible involvement of semi-mature DCs in Th17 cell differentiation. PMID- 19559669 TI - Ibuprofen modulates allosterically NO dissociation from ferrous nitrosylated human serum heme-albumin by binding to three sites. AB - Human serum albumin (HSA) is a monomeric allosteric protein. Here, the effect of ibuprofen on denitrosylation kinetics (k(off)) and spectroscopic properties of HSA-heme-Fe(II)-NO is reported. The k(off) value increases from (1.4+/-0.2)x10( 4)s(-1), in the absence of the drug, to (9.5+/-1.2)x10(-3)s(-1), in the presence of 1.0x10(-2)M ibuprofen, at pH 7.0 and 10.0 degrees C. From the dependence of k(off) on the drug concentration, values of the dissociation equilibrium constants for ibuprofen binding to HSA-heme-Fe(II)-NO (K(1)=(3.1+/-0.4)x10(-7)M, K(2)=(1.7+/-0.2)x10(-4)M, and K(3)=(2.2+/-0.2)x10(-3)M) were determined. The K(3) value corresponds to the value of the dissociation equilibrium constant for ibuprofen binding to HSA-heme-Fe(II)-NO determined by monitoring drug-dependent absorbance spectroscopic changes (H=(2.6+/-0.3)x10(-3)M). Present data indicate that ibuprofen binds to the FA3-FA4 cleft (Sudlow's site II), to the FA6 site, and possibly to the FA2 pocket, inducing the hexa-coordination of HSA-heme-Fe(II) NO and triggering the heme-ligand dissociation kinetics. PMID- 19559670 TI - Myocardial stunning is associated with impaired calcium uptake by sarcoplasmic reticulum. AB - Myocardial stunning (temporary post-ischaemic contractile dysfunction) may be caused by oxidative stress and/or impaired myocyte calcium homeostasis. Regional myocardial stunning was induced in open-chest pigs (segment shortening reduced to 68.3+/-4.7% of baseline) by repetitive brief circumflex coronary occlusion (I/R). Reduced glutathione was depleted in stunned myocardium (1.34+/-0.06 vs. 1.77+/ 0.11 nmol/mg, p=0.02 vs. remote myocardium) indicating regional oxidant stress, but no regional differences were observed in protein-bound 3-nitrotyrosine or S nitrosothiol content. Repetitive I/R did not affect myocardial quantities of the sarcolemmal sodium-calcium exchanger, L-type channel, SR calcium ATPase and phospholamban, or the kinetics of ligand binding to L-type channels and SR calcium release channels. However, initial rates of oxalate-supported (45)Ca uptake by SR were impaired in stunned myocardium (41.3+/-13.5 vs. 73.0+/-15.6 nmol/min/mg protein, p=0.03). The ability of SR calcium ATPase to sequester cytosolic calcium is impaired in stunned myocardium. This is a potential mechanism underlying contractile dysfunction. PMID- 19559671 TI - Basal nitric oxide release attenuates cell migration of HeLa and endothelial cells. AB - Nitric oxide (NO) generated by endothelial NO synthase (eNOS) is a key regulator of endothelial cell (EC) migration. Whereas the effects of acute NO generation are generally stimulatory, the role of chronic basal NO release has not been explored so far. Here, we addressed this question both in HeLa and in human endothelial cells. In stably transfected HeLa cells, inducibly expressing eNOS, expression of the enzyme per se blunted the phosphorylation of Akt/PKB in response to serum and strongly inhibited chemotaxis, an effect partially blocked by eNOS- and soluble guanylyl cyclase (sGC) inhibitors. Likewise, long-term pre treatment of non-transfected HeLa cells with nanomolar concentrations of an NO donor inhibited subsequent migration, an effect blocked by sGC inhibition and mimicked by a cGMP analog. Finally, EC migration was stimulated by chronic pre treatment with an eNOS inhibitor. Thus, in addition to its well-known stimulatory role, eNOS attenuates migration through basal long-term NO release. PMID- 19559672 TI - Tumor necrosis factor-alpha enhances IL-15-induced natural killer cell differentiation. AB - The differentiation of natural killer (NK) cells is regulated by various factors including soluble growth factors and transcription factors. Here, we have demonstrated that tumor necrosis factor-alpha (TNF-alpha) is a positive regulator of NK cell differentiation. TNF-alpha augmented the IL-15-induced expression of NK1.1 and CD122 in mature NK cells, and TNF-alpha alone also induced NK cell maturation as well as IL-15. TNF-alpha also increased IFN-gamma production in NK cells in the presence of IL-15. Meanwhile, mRNA expression of several transcription factors, including T-bet and GATA-3, was increased by the addition of TNF-alpha and IL-15. In addition, TNF-alpha increased nuclear factor-kappa B (NF-kappaB) activity in NK cells and inhibition of NF-kappaB impeded TNF-alpha enhanced NK cell maturation. Overall, these data suggest that TNF-alpha significantly increased IL-15-driven NK cell differentiation by increasing the expression of transcription factors that play crucial roles in NK cell maturation and inducing the NF-kappaB activity. PMID- 19559673 TI - CD47 promotes both phosphatidylserine-independent and phosphatidylserine dependent phagocytosis of apoptotic murine thymocytes by non-activated macrophages. AB - The ubiquitously expressed cell surface glycoprotein CD47 on host cells can inhibit phagocytosis of unopsonized or opsonized viable host target cells. Here we studied the role of target cell CD47 in macrophage uptake of viable or apoptotic murine thymocytes. As expected, IgG-opsonized viable CD47(-/-) thymocytes were taken up more efficiently than equally opsonized Wt thymocytes. However IgG-opsonized apoptotic thymocytes from Wt and CD47(-/-) mice were taken up equally. Although uptake of apoptotic thymocytes by non-activated bone marrow derived macrophages was phosphatidylserine (PS)-independent, while uptake by non activated resident peritoneal macrophages was PS-dependent, both macrophage populations showed a reduced uptake of non-opsonized apoptotic CD47(-/-) thymocytes, as compared with the uptake of apoptotic Wt thymocytes. This difference was only seen with non-activated macrophages, and not with beta-1,3 glucan-activated macrophages. CD47 promoted binding of thymocytes to macrophages, which did not require F-actin polymerization. CD47 became clustered on apoptotic thymocytes, both co-localized with or separated from, clustered PS and cholesterol-rich GM-1 domains. Thus, CD47 does not inhibit, but rather support, both PS-independent and PS-dependent uptake of apoptotic cells in the murine system. This mechanism only comes into play in non-activated macrophages. PMID- 19559674 TI - Identification of a small molecule SIRT2 inhibitor with selective tumor cytotoxicity. AB - As a member of the class III histone deacetylases, Sirtuin-2 (SIRT2) is critical in cell cycle regulation which makes it a potential target for cancer therapeutics. In this study, we identified a novel SIRT2 inhibitor, AC-93253, with IC(50) of 6 microM in vitro. The compound is selective, inhibiting SIRT2 7.5 and 4-fold more potently than the closely related SIRT1 and SIRT3, respectively. AC-93253 significantly enhanced acetylation of tubulin, p53, and histone H4, confirming SIRT2 and SIRT1 as its cellular targets. AC-93253 as a single agent exhibited submicromolar selective cytotoxicity towards all four tumor cell lines tested with a therapeutic window up to 200-fold, comparing to any of the three normal cell types tested. Results from high content analysis suggested that AC 93253 significantly triggered apoptosis. Taken together, SIRT2 selective inhibitor AC-93253 may serve as a novel chemical scaffold for structure-activity relationship study and future lead development. PMID- 19559675 TI - Telomerase reverse transcriptase activates the expression of vascular endothelial growth factor independent of telomerase activity. AB - Telomerase is almost universally required for cellular immortality and is permissive for tumorigenesis. Accumulating evidence suggests that telomerase may have roles in cellular processes independent of its role in telomere maintenance. However, the biochemical properties and the mechanisms of telomerase non telomeric functions are unknown. In this report, we show that the telomerase catalytic component hTERT is capable of activating transcription of VEGF. We further demonstrated that the activation of VEGF expression by hTERT was independent of telomerase activity and telomere maintenance. These results suggest that hTERT has non-telomeric activity in the regulation of gene expression and provide additional insights into the role of telomerase in aging and cancer. PMID- 19559677 TI - Regulator of G-protein signaling (RGS) proteins in cancer biology. AB - The regulator of G-protein signaling (RGS) family is a diverse group of multifunctional proteins that regulate cellular signaling events downstream of G protein coupled receptors (GPCRs). In recent years, GPCRs have been linked to the initiation and progression of multiple cancers; thus, regulators of GPCR signaling are also likely to be important to the pathophysiology of cancer. This review highlights recent studies detailing changes in RGS transcript expression during oncogenesis, single nucleotide polymorphisms in RGS proteins linked to lung and bladder cancers, and specific roles for RGS proteins in multiple cancer types. PMID- 19559676 TI - Knockout of the regulatory factor X1 gene leads to early embryonic lethality. AB - The biological function of regulatory factor X1 (RFX1), the prototype member of the transcription factor RFX family, is not clear. We have used gene trap technique to disrupt the expression of RFX1 in mice. Although, heterozygous RFX1(+/-) mice appear normal and fertile, homozygous RFX1(-/-) embryos died at an early stage (most likely before embryonic day 2.5). Our results indicate that RFX1 regulates expression of genes that are essential for early embryonic development/survival and that RFX1 function can not be compensated by other RFX1 family members. PMID- 19559678 TI - Odoroside A and ouabain inhibit Na+/K+-ATPase and prevent NF-kappaB-inducible protein expression by blocking Na+-dependent amino acid transport. AB - Inflammatory cytokines, such as tumor necrosis factor (TNF)-alpha and interleukin 1 (IL-1), trigger the activation of transcription factor NF-kappaB that induces the expression of a variety of genes, including intercellular adhesion molecule (ICAM)-1. Odoroside A [3beta-O-(beta-D-diginosyl)-14-hydroxy-5beta,14beta-card 20(22)-enolide] was found to inhibit the cell-surface expression of ICAM-1 induced by TNF-alpha and IL-1 at comparable concentrations in human lung carcinoma A549 cells. In this study, the molecular mechanism underlying the inhibition of TNF-alpha-induced cell-surface ICAM-1 expression by odoroside A together with the specific Na(+)/K(+)-ATPase inhibitor ouabain was further investigated. Odoroside A and ouabain neither prevented IkappaBalpha degradation nor NF-kappaB translocation to the nucleus upon TNF-alpha stimulation. While odoroside A and ouabain had no inhibitory effect on the induction of ICAM-1 mRNA, they inhibited the TNF-alpha-induced ICAM-1 expression at the protein level. Consistent with these results, odoroside A and ouabain potently reduced de novo protein synthesis, largely due to its ability to block Na(+)-dependent transport of amino acids across the plasma membrane, but not to interfering with the translation machinery. As a direct molecular target, odoroside A was found to inhibit the ATP-hydrolyzing activity of Na(+)/K(+)-ATPase as potently as ouabain. These results clearly demonstrate that odoroside A and ouabain prevent NF-kappaB inducible protein expression by blocking the Na(+)-dependent amino acid transport. PMID- 19559679 TI - The role of CC chemokine receptor 2 on microglia activation and blood-borne cell recruitment after transient focal cerebral ischemia in mice. AB - The chemokine receptor 2 (CCR2) is involved in inflammatory reactions following cerebral ischemia. Monocyte chemoattractant protein-1 (MCP-1) binds with high affinity to CCR2. MCP-1 is necessary for recruiting blood-borne cells to the injury site whereas it does not affect microglia activation and migration. MCP-1 deficient mice develop smaller infarcts and show a better functional outcome. CCR2-deficient mice also develop smaller infarcts and have a reduced expression of inflammatory cytokines during reperfusion. In the present study we investigated the differential role of inflammatory cells in CCR2-deficient mice, using green fluorescent protein (GFP)-transgenic bone marrow chimeras. After 30 min of transient middle cerebral artery occlusion (MCAO), activation of local microglia was similar in CCR2-deficient animals and their littermate controls over the study period, whereas an influx of GFP-positive cells was diminished in CCR2-deficient mice. Infiltrating macrophages were significantly reduced at day seven in the deficient animals (26.04+/-25.19 cells/mm(2)) compared to control mice (86.83+/-44.41 cells/mm(2), p<0.001). Neutrophils were also significantly reduced in CCR2-deficient mice (83% on day 2, 76% on day 4 and 89% on day 7, p<0.001). A significant reduction of infarct volume in CCR2-deficient animals could not be detected. In this study a clear differentiation of local and blood borne inflammatory cell reaction after cerebral ischemia could be shown, demonstrating that CCR2-deficiency attenuates hematogenous cell recruitment to the injury site whereas microglia activation and migration is not affected. PMID- 19559680 TI - Vestibular afferents to the motoneurons of glossopharyngeal and vagus nerves in the frog, Rana esculenta. AB - The aim of this work was to study whether the vestibular afferent fibers establish direct connections with the motoneurons of glossopharyngeal and vagus nerves of the frog, Rana esculenta. In anaesthetized animals the vestibulocochlear nerve and the common root of glossopharyngeal-vagus and accessory (IX-X-XI) nerves were simultaneously labeled with fluorescein dextran amine (vestibulocochlear nerve) and tetramethylrhodamine dextran amine (IX-X-XI). With a confocal laser scanning microscope we could detect close appositions between the vestibular afferent fibers and somatodendritic components of the general and special visceral motoneurons of the ambiguus nucleus of IX-X nerves. The direct impulse transmission may provide a quick and immediate response of cardiovascular and gastrointestinal system upon body displacement. PMID- 19559681 TI - Unilateral cortical spreading depression induced by sound in rats. AB - Cortical spreading depression (SD) is thought to underlie the migraine aura but the mechanisms of triggering SD in the human cortex remain unknown. Because growing evidence points to a key role of brainstem circuits in initiating migraine attacks, the present study examined whether recurrent episodes of brainstem activation in rats could induce cortical SD. Explosive running elicited by sounds in rodents with inherited hypersensitivity to acoustic stimuli (reflex audiogenic epilepsy), is known to reflect a transient aberrant activation of the brainstem. Repeated induction of such audiogenic responses enhances the excitability of the cortex, culminating in its epileptic activation (audiogenic kindling). In Wistar rats with inherited hypersensitivity to sounds, 15 brief episodes of running were induced by sound stimulation, and slow potential shifts as well as the EEG were recorded in the cortex. Single unilateral SD began to occur in the cortex following a running episode after the 5th to 15th test (mean 9.4+/-1.2). Once appeared, SD was regularly recorded in subsequent tests. The side of the SD initiation closely correlated with the direction of running. Triggering SD was not associated with epileptic activation of the cortex in most rats. The present findings suggest that the sensory-induced brainstem excitation could be a potent trigger of SD in the hyperexcitable cortex, providing an experimental evidence of a possible causative role of the brainstem activation in initiating the migraine aura. PMID- 19559682 TI - Heterogeneity of the infant stage of rat development: inflammatory pain response, depression-related behavior, and effects of prenatal stress. AB - The infant stage of rat development is a very important period for potential correction of adverse consequences produced by negative prenatal events. However the age limit for this correction needs to be investigated. The last prenatal and first two weeks after birth are "critical" for maturation of the nociceptive and emotional systems. Clinical observations suggest a correlation between persistent pain response and emotional behavior. In infant male rats of different ages, we studied indices of the inflammatory pain response (the number of flexes+shakes in the formalin test), depression-related behavior (immobility in the forced swim test) and the relations between them, as well as the effects of prenatal stress on these indices. Furthermore, we assessed the trend of body weight and the relations between body weight and the depression- and pain-related behaviors. We demonstrate heterogeneity of the infant stage: control prenatally non-stressed rat pups showed significantly lower immobility at 7 days of age than at 10 days; prenatal stress caused an increase of immobility and the number of flexes+shakes in 7-8-day-old pups but not in 10-11-day-olds. These findings should be taken into account in the treatment of abnormalities of emotional and inflammatory pain related behaviors produced by prenatal stressful events. The present data and our previous findings indicate that the deficiency of body weight in prenatally stressed newborns may predict the development of abnormalities in inflammatory pain-related responses during postnatal ontogeny. PMID- 19559683 TI - Presynaptic actions of propofol enhance inhibitory synaptic transmission in isolated solitary tract nucleus neurons. AB - General anesthetics variably enhance inhibitory synaptic transmission that relies on (-aminobutyric acid (GABA) and GABAA receptor function with distinct differences across brain regions. Activation of "extra-synaptic" GABAA receptors produces a tonic current considered the most sensitive target for general anesthetics, particularly in forebrain neurons. To evaluate the contribution of poor drug access to neurons in slices, we tested the intravenous anesthetic propofol in mechanically isolated neurons from the solitary tract nucleus (NTS). Setting chloride concentrations to ECl=-29 mV made GABA currents inward at holding potentials of -60 mV. Propofol triggered pronounced but slowly-developing tonic currents that reversed with 5 min washing. Effective concentrations in isolated cells were lower than in slices and propofol enhanced phasic IPSCs more potently than tonic currents (1 microM increased phasic decay-time constant vs. >3 microM tonic currents). Propofol increased IPSC frequency (>3 microM), a presynaptic action. Bicuculline blocked all propofol actions. Gabazine blocked only phasic IPSCs. IPSCs persisted in TTX and/or cadmium but these agents prevented propofol-induced increases in IPSC frequency. Furosemide (>1 mM) reversibly blocked propofol-evoked IPSC frequency changes without altering waveforms. We conclude that presynaptic actions of propofol depend on a depolarizing chloride gradient across presynaptic inhibitory terminals. Our results in isolated neurons indicate that propofol pharmacokinetics intrinsically trigger the tonic currents slowly and the time course is not related to slow permeation or delivery. Unlike forebrain, phasic NTS GABAA receptors are more sensitive to propofol than tonic receptors but that presynaptic GABAA receptor mechanisms regulate GABA release. PMID- 19559685 TI - Human brain responsivity to different intensities of masked fearful eye whites: an ERP study. AB - Previous studies have shown differential event-related potentials (ERPs) to intensities of fearful facial expressions. There are indications that the eyes may be particularly relevant for the recognition of fearful expressions, even the amount of white sclera exposed above and on sides of the dark pupil could activate the amygdala response. To investigate whether the ERP differences between intensities of fearful expressions are driven by the differential salience of the eyes in the fearful faces, ERPs were measured within a backward masking paradigm, where observers were asked to do a gender-decision task with male and female neutral faces. The emotional stimuli used were low-intensity (50%), prototypical (100%), and caricatured (150%) fearful eye whites that were derived from corresponding intensities of fearful faces respectively. Three groups of white squares that have the same pixels as the eye whites were created as control conditions. Analysis of the ERP data showed a linear increase in amplitudes of the parietal-occipital P120 by three intensities of fearful eye whites. These ERP effects were proved sensitive to intensities of negative emotions but not to the simple physical features as the same patterns of differences were not observed on white squares. Larger parietal-occipital P250 amplitudes were observed for caricatured 150% than low-intensity 50% fearful eye whites. It might reflect the subcortical pathway of emotion-specific, fearful processing. The results demonstrate that the human brain is sensitive to intensities of fear, even if just shown intensities of fearful eye white in the absence of awareness. PMID- 19559686 TI - Importance of NOX1 for angiotensin II-induced cerebrovascular superoxide production and cortical infarct volume following ischemic stroke. AB - Angiotensin II (Ang II) receptor blockade is beneficial in stroke, possibly due to attenuation of vascular oxidative stress. Mice genetically targeted for the superoxide-forming vascular NADPH oxidase subunit, NOX1, have a blunted hypertensive response to Ang II. We therefore hypothesised that NOX1 is mechanistically involved in Ang II-induced superoxide production by cerebral arteries, and potentially in stroke outcome. Superoxide production by cerebral arteries and brains from wild-type (WT) and NOX1 deficient (NOX1-KO) mice was measured using L-012-enhanced chemiluminescence. Ischemic stroke was induced by middle cerebral artery occlusion (MCAO; 0.5 h). Cerebral blood flow was measured using transcranial laser-Doppler flowmetry. After 24 h, neurological assessment was performed, mice were euthanised, and infarct and edema volumes were calculated. Basal superoxide was similar between WT and NOX1-KO in brain and cerebral artery homogenates, and in intact cerebral arteries. However, Ang II stimulated increases in superoxide were approximately 70% smaller in rings from NOX1-KO versus WT. During MCAO, rCBF decreased by approximately 75% in both WT and NOX1-KO, and increased to similar levels in each strain immediately following reperfusion. No difference in neurological score, total or subcortical cerebral infarct volume or edema volume was observed between WT and NOX1-KO mice. However, cortical infarct volume (which was very modest in WT) was approximately 4-fold greater in brains of NOX1-KO versus WT. Thus, NOX1 is essential for superoxide production in large cerebral arteries in response to Ang II but not under basal conditions. Furthermore, NOX1 does not appear to contribute to stroke size, and it may limit cortical infarct development following cerebral ischemia. PMID- 19559684 TI - Acupuncture mobilizes the brain's default mode and its anti-correlated network in healthy subjects. AB - Previous work has shown that acupuncture stimulation evokes deactivation of a limbic-paralimbic-neocortical network (LPNN) as well as activation of somatosensory brain regions. This study explores the activity and functional connectivity of these regions during acupuncture vs. tactile stimulation and vs. acupuncture associated with inadvertent sharp pain. Acupuncture during 201 scans and tactile stimulation during 74 scans for comparison at acupoints LI4, ST36 and LV3 was monitored with fMRI and psychophysical response in 48 healthy subjects. Clusters of deactivated regions in the medial prefrontal, medial parietal and medial temporal lobes as well as activated regions in the sensorimotor and a few paralimbic structures can be identified during acupuncture by general linear model analysis and seed-based cross correlation analysis. Importantly, these clusters showed virtual identity with the default mode network and the anti correlated task-positive network in response to stimulation. In addition, the amygdala and hypothalamus, structures not routinely reported in the default mode literature, were frequently involved in acupuncture. When acupuncture induced sharp pain, the deactivation was attenuated or became activated instead. Tactile stimulation induced greater activation of the somatosensory regions but less extensive deactivation of the LPNN. These results indicate that the deactivation of the LPNN during acupuncture cannot be completely explained by the demand of attention that is commonly proposed in the default mode literature. Our results suggest that acupuncture mobilizes the anti-correlated functional networks of the brain to mediate its actions, and that the effect is dependent on the psychophysical response. PMID- 19559687 TI - Ketogenic diet protects dopaminergic neurons against 6-OHDA neurotoxicity via up regulating glutathione in a rat model of Parkinson's disease. AB - The high-fat ketogenic diet (KD) leads to an increase of blood ketone bodies (KB) level and has been used to treat refractory childhood seizures for over 80 years. Recent reports show that KD, KB and their components (d-beta-hydroxybutyrate, acetoacetate and acetone) have neuroprotective for acute and chronic neurological disorders. In our present work, we examined whether KD protected dopaminergic neurons of substantia nigra (SN) against 6-hydroxydopamine (6-OHDA) neurotoxicity in a rat model of Parkinson's disease (PD) using Nissl staining and tyrosine hydroxylase (TH) immunohistochemistry. At the same time we measured dopamine (DA) and its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the striatum. To elucidate the mechanism, we also measured the level of glutathione (GSH) of striatum. Our data showed that Nissl and TH-positive neurons increased in rats fed with KD compared to rats with normal diet (ND) after intrastriatal 6-OHDA injection, so did DA and its metabolite DOPAC. While HVA had not changed significantly. The change of GSH was significantly similar to DA. We concluded that KD had neuroprotective against 6-OHDA neurotoxicity and in this period GSH played an important role. PMID- 19559688 TI - The medial hypothalamic defensive circuit and 2,5-dihydro-2,4,5 trimethylthiazoline (TMT) induced fear: comparison of electrolytic and neurotoxic lesions. AB - The neural circuits for unconditioned fear to predator odors (e.g., cat fur odor, trimethylthiazoline, TMT) are not well delineated. A putative neural circuit for predator odor fear, the medial hypothalamic defensive circuit (MHDC), consisting of the anterior hypothalamic (AHN), ventromedial hypothalamic (VMH) and dorsal premammillary nuclei (PMd), has been proposed. Electrolytic and ibotenic acid lesions of the PMd have been shown to reduce unconditioned fear in rats presented with either a cat or cat odor. Whether the PMd, AHN and VMH are involved in unconditioned fear to another predator odor derived from fox feces, 2,5-dihydro 2,4,5-trimethylthiazoline (TMT), has not been explored. The present study compared the effects of electrolytic and neurotoxic lesions of MHDC nuclei in rats on unconditioned fear to TMT and shock-induced contextually conditioned fear, as measured by freezing. Electrolytic lesions of the PMd did not reduce TMT induced freezing, but diminished post-shock and shock-induced contextually conditioned freezing, suggesting a role for the PMd in contextually conditioned fear. In contrast, electrolytic lesions of the AHN and VMH reduced freezing to TMT while not affecting conditioned fear. However, neither NMDA lesions of the AHN nor ibotenic acid lesions of the VMH reduced freezing in shock-induced conditioned or TMT-induced unconditioned fear paradigms. The data suggest that fibers passing through the AHN and VMH, and not cells in the MHDC, mediate unconditioned freezing to the predator odor TMT. PMID- 19559689 TI - Tissue persistence of fumonisin B1 in ducks and after exposure to a diet containing the maximum European tolerance for fumonisins in avian feeds. AB - Toxicity and persistence of fumonisin B1 (FB1) in liver, kidney and muscle were investigated in ducks fed 5, 10 and 20mg FB1+FB2/kg feed during force-feeding. Mortality and signs of toxicity were only obtained with 20mg/kg, whereas an increased Sa/So ratio was observed from 5mg/kg on. Persistence of FB1 was only found in liver (16 and 20 microg FB1/kg liver in ducks fed 10 and 20 mg FB1+FB2/kg feed, respectively). Toxicokinetic studies were conducted by the intravenous route (IV, single dose: 10mg FB1/kg body weight) and the oral route (single dose: 100mg FB1/kg body weight), in growing ducks and in ducks during force-feeding. After IV administration, serum concentration-time curves were described by a two-compartment open model. Elimination half-life and mean residence time of FB1 were 26 and 24 min, respectively, clearance was 19.3 ml/min/kg. After oral administration, bioavailability, elimination half-life, mean residence time and clearance varied during force-feeding and growth from 2 2.3%, 71-80 min, 200-188 min, 16.7-17 ml/min/kg, respectively. Taken together these results demonstrate that the risk of persistence of FB1 in ducks after force-feeding is very low, Sa/So being a good biomarker which increases before signs of toxicity and risk of persistence of FB1 in tissue (limit of detection 13 microg/kg). PMID- 19559690 TI - Lymphocyte transformation test is not helpful for the diagnosis of methotrexate induced pneumonitis in patients with rheumatoid arthritis. AB - BACKGROUND: The pathophysiology underlying methotrexate (MTX)-induced pneumonitis has been considered as a hypersensitive reaction. The lymphocyte transformation test (LTT) is frequently used to detect hypersensitivity. Whereas previous reports have proposed that the LTT is not ideal to detect hypersensitivity to MTX, it has not been directly confirmed. METHODS: Forty rheumatoid arthritis (RA) patients (24 patients currently taking MTX and 16 patients with a past history of MTX administration) and 13 healthy subjects were recruited. LTT with MTX was used to assess thymidine incorporation. An MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) assay was also performed. The mitogenic activity was expressed as the Stimulatory Index (SI). The activity of RA was assessed by the disease activity score 28 (DAS28). RESULTS: In the presence of MTX, the SI measured by the LTT and by the MTS assay showed an inverse correlation. The presence of MTX significantly elevated the SI values measured by the LTT. However, the SI values were significantly lower in RA patients currently taking MTX than those of patients not currently taking MTX, although DAS28 was not different. Furthermore, a past history of MTX-induced pneumonitis did not affect the SI values. CONCLUSION: LTT with MTX in RA patients is not appropriate to detect MTX-induced pneumonitis. PMID- 19559691 TI - Quantitative UPLC-MS/MS analysis of underivatised amino acids in body fluids is a reliable tool for the diagnosis and follow-up of patients with inborn errors of metabolism. AB - BACKGROUND: An electro-spray ionisation ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) application for the quantitative analysis of amino acids was developed. The suitability for the detection and follow-up of patients suffering from inborn errors of metabolism (IEM) was assessed by extensive cross-validation with ion-exchange liquid chromatography (IEX-LC) with post-column ninhydrin derivatisation, participation in external quality control (ERNDIM) and analysis of samples of patients with confirmed IEM. METHODS: Prior to analysis plasma and urine samples were merely diluted 150-fold in mobile phase. Amino acids were detected in the multiple reaction monitoring mode (MRM) in the ESI-positive mode. The analytical results were compared with IEX-LC. External quality control scheme performance is presented. RESULTS: Comprehensive analysis of amino acids in plasma and urine was achieved with a run-to-run time of 30min. Validation results were satisfactory and there was a very good correlation between UPLC-MS/MS and IEX-LC. Analytical results obtained in the external quality control scheme were essentially the same as those of the other participants. Patients suffering from IEM were readily identified. CONCLUSION: UPLC-MS/MS analysis of amino acids in body fluids is rapid, reliable and suitable for the diagnosis and follow-up patients with IEM. PMID- 19559692 TI - Isochoric preservation: a novel characterization method. AB - Isochoric (constant volume) preservation is an alternative to traditional cryopreservation methods because it requires less cryoprotectant and is simple to operate. In order to validate that this method automatically minimizes the pressure for a given temperature, pressure and temperature data were collected from a specially designed pressure vessel. This vessel was then used to examine the effect of an isochoric environment on freezing point nucleation in an aqueous antifreeze protein solution, and to generate pressure-temperature phase diagrams for various cryoprotectant solutions. Our results show that the isochoric pressure vessel follows the pressure-temperature phase diagram of water, thereby minimizing the pressure for the given temperature. We also show that the nucleation temperature of the antifreeze protein in an isochoric vessel is lower than that of the isobaric method. Furthermore, the nucleation temperature decreased with increasing concentration in the isochoric vessel while the isobaric nucleation temperature showed no change. These results indicate that the isochoric environment imposes additional constraints on ice formation and warrants further study as these results may change when a different type of cryoprotectant is used. Finally, all of the cryoprotectant phase diagrams exhibited a similar pressure-temperature slope indicating that, regardless of the cryoprotectant used or the mechanism by which it suppresses freezing, isochoric freezing affects the molecules in the same manner. Together, all of these results indicate that the isochoric method of preservation is a valuable tool for characterizing the thermodynamic properties of cryoprotectants and has great potential as a cryopreservation method in the field of cryobiology. PMID- 19559693 TI - The proprotein convertase amontillado (amon) is required during Drosophila pupal development. AB - Peptide hormones governing many developmental processes are generated via endoproteolysis of inactive precursor molecules by a family of subtilisin-like proprotein convertases (SPCs). We previously identified mutations in the Drosophila amontillado (amon) gene, a homolog of the vertebrate neuroendocrine specific Prohormone Convertase 2 (PC2) gene, and showed that amon is required during embryogenesis, early larval development, and larval molting. Here, we define amon requirements during later developmental stages using a conditional rescue system and find that amon is required during pupal development for head eversion, leg and wing disc extension, and abdominal differentiation. Immuno localization experiments show that amon protein is expressed in a subset of central nervous system cells but does not co-localize with peptide hormones known to elicit molting behavior, suggesting the involvement of novel regulatory peptides in this process. The amon protein is expressed in neuronal cells that innervate the corpus allatum and corpora cardiaca of the ring gland, an endocrine organ which is the release site for many key hormonal signals. Expression of amon in a subset of these cell types using the GAL4/UAS system in an amon mutant background partially rescues larval molting and growth. Our results show that amon is required for pupal development and identify a subset of neuronal cell types in which amon function is sufficient to rescue developmental progression and growth defects shown by amon mutants. The results are consistent with a model that the amon protein acts to proteolytically process a diverse suite of peptide hormones that coordinate larval and pupal growth and development. PMID- 19559695 TI - Sailing with the Wnt: charting the Wnt processing and secretion route. AB - Wnt proteins are members of a highly conserved family of signalling molecules that play a central role in development and disease. During the past years, the different signalling pathways that are triggered by Wnt proteins have been studied in detail, but it is still largely unknown how a functional Wnt protein is produced and secreted. The recent finding that Wnt proteins are post translationally modified and the discovery of the Wnt binding protein Wntless and its trafficking by the retromer complex show that Wnt secretion is a complex and highly regulated process. In this review, we will give an overview of the Wnt maturation and secretion pathway and discuss how this process may influence the spreading and signalling activity of Wnt. PMID- 19559694 TI - miR-17 family of microRNAs controls FGF10-mediated embryonic lung epithelial branching morphogenesis through MAPK14 and STAT3 regulation of E-Cadherin distribution. AB - The miR-17 family of microRNAs has recently been recognized for its importance during lung development. The transgenic overexpression of the entire miR-17-92 cluster in the lung epithelium led to elevated cellular proliferation and inhibition of differentiation, while targeted deletion of miR-17-92 and miR-106b 25 clusters showed embryonic or early post-natal lethality. Herein we demonstrate that miR-17 and its paralogs, miR-20a, and miR-106b, are highly expressed during the pseudoglandular stage and identify their critical functional role during embryonic lung development. Simultaneous downregulation of these three miRNAs in explants of isolated lung epithelium altered FGF10 induced budding morphogenesis, an effect that was rescued by synthetic miR-17. E-Cadherin levels were reduced, and its distribution was altered by miR-17, miR-20a and miR-106b downregulation, while conversely, beta-catenin activity was augmented, and expression of its downstream targets, including Bmp4 as well as Fgfr2b, increased. Finally, we identified Stat3 and Mapk14 as key direct targets of miR-17, miR-20a, and miR 106b and showed that simultaneous overexpression of Stat3 and Mapk14 mimics the alteration of E-Cadherin distribution observed after miR-17, miR-20a, and miR 106b downregulation. We conclude that the mir-17 family of miRNA modulates FGF10 FGFR2b downstream signaling by specifically targeting Stat3 and Mapk14, hence regulating E-Cadherin expression, which in turn modulates epithelial bud morphogenesis in response to FGF10 signaling. PMID- 19559697 TI - Hedgehog signal activation coordinates proliferation and differentiation of fetal liver progenitor cells. AB - Hedgehog (Hh) signaling plays crucial roles in development and homeostasis of various organs. In the adult liver, it regulates proliferation and/or viability of several types of cells, particularly under injured conditions, and is also implicated in stem/progenitor cell maintenance. However, the role of this signaling pathway during the normal developmental process of the liver remains elusive. Although Sonic hedgehog (Shh) is expressed in the ventral foregut endoderm from which the liver derives, the expression disappears at the onset of the liver bud formation, and its possible recurrence at the later stages has not been investigated. Here we analyzed the activation and functional relevance of Hh signaling during the mouse fetal liver development. At E11.5, Shh and an activation marker gene for Hh signaling, Gli1, were expressed in Dlk(+) hepatoblasts, the fetal liver progenitor cells, and the expression was rapidly decreased thereafter as the development proceeded. In the culture of Dlk(+) hepatoblasts isolated from the E11.5 liver, activation of Hh signaling stimulated their proliferation and this effect was cancelled by a chemical Hh signaling inhibitor, cyclopamine. In contrast, hepatocyte differentiation of Dlk(+) hepatoblasts in vitro as manifested by the marker gene expression and acquisition of ammonia clearance activity was significantly inhibited by forced activation of Hh signaling. Taken together, these results demonstrate the temporally restricted manner of Hh signal activation and its role in promoting the hepatoblast proliferation, and further suggest that the pathway needs to be shut off for the subsequent hepatic differentiation of hepatoblasts to proceed normally. PMID- 19559696 TI - Mesenchymal to embryonic incomplete transition of human cells by chimeric OCT4/3 (POU5F1) with physiological co-activator EWS. AB - POU5F1 (more commonly known as OCT4/3) is one of the stem cell markers, and affects direction of differentiation in embryonic stem cells. To investigate whether cells of mesenchymal origin acquire embryonic phenotypes, we generated human cells of mesodermal origin with overexpression of the chimeric OCT4/3 gene with physiological co-activator EWS (product of the EWSR1 gene), which is driven by the potent EWS promoter by translocation. The cells expressed embryonic stem cell genes such as NANOG, lost mesenchymal phenotypes, and exhibited embryonal stem cell-like alveolar structures when implanted into the subcutaneous tissue of immunodeficient mice. Hierarchical analysis by microchip analysis and cell surface analysis revealed that the cells are subcategorized into the group of human embryonic stem cells and embryonal carcinoma cells. These results imply that cells of mesenchymal origin can be traced back to cells of embryonic phenotype by the OCT4/3 gene in collaboration with the potent cis-regulatory element and the fused co-activator. The cells generated in this study with overexpression of chimeric OCT4/3 provide us with insight into cell plasticity involving OCT4/3 that is essential for embryonic cell maintenance, and the complexity required for changing cellular identity. PMID- 19559698 TI - Chemokines released from astrocytes promote chemokine receptor 5-mediated neuronal cell differentiation. AB - Astrocytes are one of major glial cell types in the central nervous system (CNS), and can support many functions of neuronal cells. In the present study, we demonstrated that the differentiation of rat embryonic neuronal cells was promoted by treatment with astrocyte and microglia-conditioned medium. Cytokine assays identified that the IL-4, MIP-1, KC, and RANTES as were released from astrocyte, and these chemokines promote differentiation of rat embryonic neuronal cells. However, chemokine-promoted neuronal cell differentiation was suppressed by antibodies of these chemokines and their receptor (CCR5). CCR5 and neuronal cell differentiation marker proteins were found to be colocalized, and their expressions were enhanced by chemokines. Furthermore, the differentiation of neuronal cells from CCR5 knock-out mice and of neuronal cells from mice knocked down with the CCR5 siRNA were significantly reduced and delayed. Bradykinin elevated calcium influx in the embryonic neuronal cells. These data suggest that specific chemokines derived from astrocytes may significantly have influence on the CCR5-mediated differentiation of embryonic neuronal cells. PMID- 19559701 TI - Muscle as a therapeutic target in amyotrophic lateral sclerosis. PMID- 19559700 TI - Somatostatin, tau, and beta-amyloid within the anterior olfactory nucleus in Alzheimer disease. AB - Impaired olfaction is an early symptom of Alzheimer disease (AD). This likely to reflect neurodegenerative processes taking place in basal telencephalic structures that mediate olfactory processing, including the anterior olfactory nucleus. Betaeta-amyloid (Abeta) accumulation in AD brain may relate to decline in somatostatin levels: somatostatin induces the expression of the Abeta degrading enzyme neprilysin and somatostatin deficiency in AD may therefore reduce Abeta clearance. We have investigated the expression of somatostatin in the anterior olfactory nucleus of AD and control brain. We report that somatostatin levels were reduced by approximately 50% in AD brain. Furthermore, triple-immunofluorescence revealed co-localization of somatostatin expression with Abeta (65.43%) with Abeta and tau (19.75%) and with tau (2.47%). These data indicate that somatostatin decreases in AD and its expression may be linked with Abeta deposition. PMID- 19559699 TI - Bilateral cervical contusion spinal cord injury in rats. AB - There is increasing motivation to develop clinically relevant experimental models for cervical SCI in rodents and techniques to assess deficits in forelimb function. Here we describe a bilateral cervical contusion model in rats. Female Sprague-Dawley rats received mild or moderate cervical contusion injuries (using the Infinite Horizons device) at C5, C6, or C7/8. Forelimb motor function was assessed using a grip strength meter (GSM); sensory function was assessed by the von Frey hair test; the integrity of the corticospinal tract (CST) was assessed by biotinylated dextran amine (BDA) tract tracing. Mild contusions caused primarily dorsal column (DC) and gray matter (GM) damage while moderate contusions produced additional damage to lateral and ventral tissue. Forelimb and hindlimb function was severely impaired immediately post-injury, but all rats regained the ability to use their hindlimbs for locomotion. Gripping ability was abolished immediately after injury but recovered partially, depending upon the spinal level and severity of the injury. Rats exhibited a loss of sensation in both fore- and hindlimbs that partially recovered, and did not exhibit allodynia. Tract tracing revealed that the main contingent of CST axons in the DC was completely interrupted in all but one animal whereas the dorsolateral CST (dlCST) was partially spared, and dlCST axons gave rise to axons that arborized in the GM caudal to the injury. Our data demonstrate that rats can survive significant bilateral cervical contusion injuries at or below C5 and that forepaw gripping function recovers after mild injuries even when the main component of CST axons in the dorsal column is completely interrupted. PMID- 19559702 TI - Identification of novel N-terminal fragments of amyloid precursor protein in cerebrospinal fluid. AB - Alzheimer's disease (AD) is a progressive neurodegenerative disorder of the central nervous system. Two pathological hallmarks in the brain of AD patients are neurofibrillary tangles and senile plaques. The plaques consist mainly of beta-amyloid (Abeta) peptides that are produced from the amyloid precursor protein (APP), by sequential cleavage by beta- and gamma-secretase. Most previous studies have been focused on the C-terminal fragments of APP, where the Abeta sequence is localized. The purpose of this study was to search for N-terminal fragments of APP in cerebrospinal fluid (CSF) using mass spectrometry (MS). By using immunoprecipitation (IP) combined with matrix-assisted laser desorption/ionization time-of-flight MS as well as nanoflow liquid chromatography coupled to high resolution tandem MS we were able to detect and identify six novel N-terminal APP fragments [APP((18-119)), APP((18-121)), APP((18-122)), APP((18-123)), APP((18-124)) and APP((18-126))], having molecular masses of approximately 12 kDa. The presence of these APP derivatives in CSF was also verified by Western blot analysis. Two pilot studies using either IP-MS or Western blot analysis indicated slightly elevated levels of N-terminal APP fragments in CSF from AD patients compared with controls, which are in need of replications in independent and larger patient materials. PMID- 19559703 TI - Mitochondrial control of autophagic lysosomal pathway in Alzheimer's disease. AB - When first described by Alois Alzheimer in 1907, AD was seen as a disorder that causes dementia and characterized by two defining neuropathological lesions, later associated with all forms of AD. While the etiology of AD remains largely unclear, there is accumulating evidence suggesting that mitochondrial dysfunction occurs prior to the onset of symptoms in AD. Mitochondria are exceptionally poised to play a crucial role in neuronal cell survival or death because they are regulators of both energy metabolism and apoptotic pathways. This review is mainly focused in the discussion of evidence suggesting a clear association between mitochondrial dysfunction, autophagy impairment and amyloid-beta accumulation in Alzheimer's disease pathophysiology. The knowledge that autophagic insufficiency may compromise the cellular degradation mechanisms that may culminate in the progressive accumulation of dysfunctional mitochondria, aberrant protein aggregates buildup and lysossomal burden shield new insights to the way we address Alzheimer's disease. In line with this knowledge an innovative window for new therapeutic strategies aimed to activate or ameliorate macroautophagy may be opened. PMID- 19559705 TI - The effect of environmental temperature on olfactory perception in Drosophila melanogaster. AB - Olfaction provides chemical information to an animal about its environment. When environmental conditions change, individuals should be able to adequately maintain function. Temperature may influence olfaction in a double manner, as it modifies the concentrations of gaseous compounds and affects biological processes. Here, we address acclimatization to environmental temperature in the olfactory system of Drosophila melanogaster using heat and cold treatments. Because the consequences of temperature shifts persist for some time after the treatment's end, comparison of olfactory behaviors at the same temperature in treated and untreated flies allows us to infer the biological effects of temperature in olfaction. At intermediate odorant concentrations heat always generates a reduction of olfactory sensitivity, as they would be expected to compensate for the increase of volatiles in the air. Cold produces the opposite effect. These changes are observed in both sexes and in natural populations as well as in standard laboratory stocks. Short applications suffice to cause detectable olfactory perception changes, but even prolonged temperature treatments have only a transitory effect. Together, these results suggest that olfaction in Drosophila underlies acclimatization to environmental temperature. However, sensitivity changes are not immediate and may cause imperfect adjustment of olfactory function for short time periods. PMID- 19559706 TI - Modulation of cyclic CO(2) release in response to endogenous changes of metabolism during pupal development of Zophobas rugipes (Coleoptera: Tenebrionidae). AB - Understanding the mechanisms of gas exchange regulation in insects currently is a hot topic of insect physiology. Endogenous variation of metabolism during pupal development offers a great opportunity to study the regulation of respiratory patterns in insects. Here we show that metabolic rates during pupal development of the tenebrionid beetle Zophobas rugipes reveal a typical U-shaped curve and that, with the exception of 9-day-old pupae, the time between two bursts of CO(2) (interburst phase) was the only parameter of cyclic CO(2) gas exchange patterns that was adjusted to changing metabolic rates. The volume of CO(2) released in a burst was kept constant, suggesting a regulation for accumulation and release of a fixed amount of CO(2) throughout pupal development. We detected a variety of discontinuous and cyclic gas exchange patterns, which were not correlated with any periods of pupal development, suggesting a high among individual variability. An occasional occurrence of continuous CO(2) release patterns at low metabolic rates was very likely caused by single defective non-occluding spiracles. PMID- 19559704 TI - Sexual experience changes sex hormones but not hypothalamic steroid hormone receptor expression in young and middle-aged male rats. AB - Testosterone is well known to regulate sexual behavior in males, but this is dependent upon prior sexual experience. Aging is associated with decreased libido and changes in testosterone, but the role of experience in these age-related processes has not been systematically studied. We examined effects of age and sexual experience on serum hormones (total testosterone, free testosterone, estradiol, LH) and on numbers of androgen receptor (AR) and estrogen receptor alpha (ERalpha) immunoreactive cells in the hypothalamus. Extensive sexual experience was given to male rats at 4 months of age. Rats were euthanized at either 4 months (young) or 12 months (middle-aged (MA)). Comparable sexually naive male rats were handled and placed into the testing arena but did not receive any sexual experience. Thus, we had four groups: young-naive, young experienced, MA-naive and MA-experienced. Serum hormone levels were assayed, and numbers of AR and ERalpha cells were quantified stereologically in the medial preoptic nucleus (MPN) and the anteroventral periventricular nucleus (AVPV). Sexually experienced males had significantly elevated serum testosterone and free testosterone in both age groups. Both total and free testosterone were higher, and estradiol lower, in middle-aged than young rats. Experience did not alter either AR or ERalpha expression in the preoptic brain regions studied. Aging was associated with increased expression of AR, but no change in ERalpha. These results show that sexual experience can induce short-term and long-term alterations in serum hormones but these effects are not manifested upon their receptors in the hypothalamus. PMID- 19559707 TI - Suppression of JH biosynthesis by JH analog treatment: mechanism of suppression and roles of allatostatins and nervous connections in the cockroach Diploptera punctata. AB - Juvenile hormone analogs are known to inhibit the production of juvenile hormone (JH) by the corpora allata (CA). However, the mechanism of this inhibition remains undefined. We have used two JH mimics, fenoxycarb and pyriproxyfen, to examine the mechanism of suppression in the cockroach, Diploptera punctata. Denervation experiments demonstrated the importance of nervous connections between the brain and CA for the inhibition of JH biosynthesis by fenoxycarb. Fenoxycarb treatment alters the sensitivity of CA to allatostatin treatment in vitro. Suppression of JH biosynthesis by fenoxycarb following denervation of the CA showed that innervation was in part responsible for the inhibition. Similarly, maximal inhibition by Dippu-AST7 requires intact nervous connections between the brain and CA, particularly during rapid vitellogenesis. qPCR analysis of brain, CA, ovary and midgut extracts revealed that both allatostatin and its receptor Dippu-ASTR2 show increased levels of expression following topical fenoxycarb treatment, particularly in brain tissue on days 4 and 5 of the first gonadotrophic cycle and in CA on day 4. The correlation between inhibition of JH biosynthesis and increased expression of AST and ASTR2 in brains and CA, together with increased sensitivity of CA to allatostatin in vitro, suggests that allatostatin may be one of the effectors by which fenoxycarb inhibits JH biosynthesis. PMID- 19559708 TI - IkB genes encoded in Cotesia plutellae bracovirus suppress an antiviral response and enhance baculovirus pathogenicity against the diamondback moth, Plutella xylostella. AB - An endoparasitoid wasp, Cotesia plutellae, parasitizes larvae of the diamondback moth, Plutella xylostella, with its symbiotic polydnavirus, C. plutellae bracovirus (CpBV). This study analyzed the role of Inhibitor-kB (IkB)-like genes encoded in CpBV in suppressing host antiviral response. Identified eight CpBV IkBs are scattered on different viral genome segments and showed high homologies with other bracoviral IkBs in their amino acid sequences. Compared to an insect ortholog (e.g., Cactus of Drosophila melanogaster), they possessed a shorter ankyrin repeat domain without any regulatory domains. The eight CpBV-IkBs are, however, different in their promoter components and expression patterns in the parasitized host. To test their inhibitory activity on host antiviral response, a midgut response of P. xylostella against baculovirus infection was used as a model reaction. When the larvae were orally fed the virus, they exhibited melanotic responses of midgut epithelium, which increased with baculovirus dose and incubation time. Parasitized larvae exhibited a significant reduction in the midgut melanotic response, compared to nonparasitized larvae. Micro-injection of each of the four CpBV genome segments containing CpBV-IkBs into the hemocoel of nonparasitized larvae showed the gene expressions of the encoded IkBs and suppressed the midgut melanotic response in response to the baculovirus treatment. When nonparasitized larvae were orally administered with a recombinant baculovirus containing CpBV-IkB, they showed a significant reduction in midgut melanotic response and an enhanced susceptibility to the baculovirus infectivity. PMID- 19559710 TI - Designing out disulfide bonds of leech carboxypeptidase inhibitor: implications for its folding, stability and function. AB - Leech carboxypeptidase inhibitor (LCI) is a 67-residue, tight-binding metallocarboxypeptidase inhibitor composed of a compact domain with a five stranded beta-sheet and a short alpha-helix that are strongly stabilized by four disulfide bonds. In this study, we investigated the contribution of each particular disulfide to the folding, stability and function of LCI by constructing a series of single and multiple mutants lacking one to four disulfide bonds. The results allow a better understanding of how individual disulfide bonds shape and restrict the conformational space that LCI must explore before attaining its native conformation. The work also dissected the role played by intramolecular rearrangements of disulfides during LCI folding, providing a new kinetic scheme in which the 2S ensemble suffers a non-specific oxidation into the 3S ensemble. These 3-disulfide-bonded species reshuffle to preferentially form III-A and III-B, two major native-like folding intermediates that need structural rearrangements through the formation of scrambled isomers to finally render native LCI. The designed multiple mutants of LCI are unable to fold correctly, displaying a highly unstructured conformation and a very low inhibitory capability, which indicates the importance of disulfide bonds in LCI for both correct folding and achievement of a functional structure. In contrast, the elimination of a single disulfide bond in LCI only results in a significant reduction of conformational stability, but the mutations have a rather moderate impact on carboxypeptidase inhibition, allowing the possibility to target the intrinsic stability and specific activity of LCI independently. In this way, the findings reported provide a basis for the design of novel variants of the molecule with improved therapeutic properties. PMID- 19559709 TI - Extracellular matrix turnover and signaling during cardiac remodeling following MI: causes and consequences. AB - The concept that extracellular matrix (ECM) turnover occurs during cardiac remodeling is a well-accepted paradigm. To date, a multitude of studies document that remodeling is accompanied by increases in the synthesis and deposition of ECM components as well as increases in extracellular proteases, especially matrix metalloproteinases (MMPs), which break down ECM components. Further, soluble ECM fragments generated from enzymatic action serve to stimulate cell behavior and have been proposed as candidate plasma biomarkers of cardiac remodeling. This review briefly summarizes our current knowledge base on cardiac ECM turnover following myocardial infarction (MI), but more importantly extends discussion by defining avenues that remain to be explored to drive the ECM remodeling field forward. Specifically, this review will discuss cause and effect roles for the ECM changes observed following MI and the potential role of the ECM changes that may serve as trigger points to regulate remodeling. While the pattern of remodeling following MI is qualitatively similar but quantitatively different from various types of injury, the basic theme in remodeling is repeated. Therefore, while we use the MI model as the prototype injury model, the themes discussed here are also relevant to cardiac remodeling due to other types of injury. PMID- 19559711 TI - The non-canonical Hop protein from Caenorhabditis elegans exerts essential functions and forms binary complexes with either Hsc70 or Hsp90. AB - Heat shock protein (Hsp) 70/Hsp90-organizing proteins (Hop/Sti1) are thought to function as adaptor proteins to link the two chaperone machineries Hsp70 and Hsp90 during the processing of substrate proteins in eukaryotes. Hop (Hsp70/Hsp90 organizing protein) is composed of three tetratricopeptide repeat (TPR) domains, of which the first (TPR1) binds to Hsp70, the second (TPR2A) binds to Hsp90, and the third (TPR2B) is of unknown function. Contrary to most other eukaryotes, the homologue closest to the Caenorhabditis elegans Hop homologue R09E12.3 (CeHop) lacks the TPR1 domain and the short linker region connecting it to TPR2A, questioning the reported function as an Hsp90/Hsp70 adaptor in vitro and in vivo. We observed high constitutive expression levels of CeHop and detected significant phenotypes upon knockdown, linking the protein to functions in gonad development. Interestingly, we observed physical interactions with both chaperones Hsp70 and Hsp90, albeit only the interaction with Hsp90 is strong and inhibition of the Hsp90 ATPase activity can be observed upon binding of CeHop. However, the formation of ternary complexes with both chaperone machineries is impaired, as Hsp70 and Hsp90 compete for CeHop interaction sites, in particular as Hsp90 binds to both TPR domains simultaneously and requires both TPR domains for ATPase regulation. These results imply that, at least in C. elegans, essential functions of Hop exist which apparently do not depend on the simultaneous binding of Hsp90 and Hsp70 to Hop. PMID- 19559712 TI - Structure of the murine unglycosylated IgG1 Fc fragment. AB - A prototypic IgG antibody can be divided into two major structural units: the antigen-binding fragment (Fab) and the Fc fragment that mediates effector functions. The IgG Fc fragment is a homodimer of the two C-terminal domains (C(H)2 and C(H)3) of the heavy chains. Characteristic of the Fc part is the presence of a sugar moiety at the inner face of the C(H)2 domains. The structure of this complex branched oligosaccharide is generally resolved in crystal structures of Fc fragments due to numerous well-defined sugar-protein interactions and a small number of sugar-sugar interactions. This suggested that sugars play an important role in the structure of the Fc fragment. To address this question directly, we determined the crystal structure of the unglycosylated Fc fragment of the murine IgG1 MAK33. The structures of the C(H)3 domains of the unglycosylated Fc fragment superimpose perfectly with the structure of the isolated MAK33 C(H)3 domain. The unglycosylated C(H)2 domains, in contrast, approach each other much more closely compared to known structures of partly deglycosylated Fc fragments with rigid-body motions between 10 and 14 A, leading to a strongly "closed" conformation of the unglycosylated Fc fragment. The glycosylation sites in the C'E loop and the BC and FG loops are well defined in the unglycosylated C(H)2 domain, however, with increased mobility and with a significant displacement of about 4.9 A for the unglycosylated Asn residue compared to the glycosylated structure. Thus, glycosylation both stabilizes the C'E-loop conformation within the C(H)2 domain and also helps to ensure an "open" conformation, as seen upon Fc receptor binding. These structural data provide a rationale for the observation that deglycosylation of antibodies often compromises their ability to bind and activate Fcgamma receptors. PMID- 19559713 TI - Memory effects on scale-free dynamics in foraging Drosophila. AB - The fruit fly, Drosophila melanogaster, displays a scale-free behavior in foraging, i.e., the dwell time on food exhibits a power law distribution. The scaling exponent is generally believed to be stable and the significance of the exponent itself with respect to the scale-free behavior remains elusive. We propose a model whereby the scaling exponent of the scale-free behavior of an animal depends on the memory of the individual. The proposed model is based on the premise that animal behaviors are associated with internal states of the animal. The changes in the scaling exponent are derived by considering losing memory as increasing uncertainty, which is expressed in terms of information entropy of the internal states. Predicted model behaviors agree with experimental results of foraging behavior in wild-type and learning/memory Drosophila mutants. The concept of changes in the scaling exponent due to the amount of memory provides a novel insight into the emergence of a scale-free behavior and the meaning of the scaling exponent. PMID- 19559714 TI - Effects of St. John's wort extract on indinavir pharmacokinetics in rats: differentiation of intestinal and hepatic impacts. AB - AIMS: To evaluate the possible herb-drug interaction of St. John's wort (SJW) extracts with indinavir in rats and to set up a model for characterizing pre systemic sites for the interactions between orally administered herbs and pharmaceuticals. MAIN METHODS: The in vivo pharmacokinetic study and in situ single-pass intestinal perfusion model were employed in the research. Plasma indinavir concentration and cytochrome P450 3A activities were measured by high pressure liquid chromatography and spectrophotometric assays, respectively. KEY FINDINGS: Oral administration of either 150 or 300 mg/day SJW for 15 days significantly reduced indinavir plasma levels with certain pharmacokinetic parameter changes. The cytochrome P450 3A analysis suggested that this interaction was attributable to the induction of indinavir metabolism. Further perfusion study demonstrated that both small intestine and the liver contributed significantly to the reduction of indinavir bioavailability and was flow rate dependent. Moreover, the small intestine was the major site for the pre-systemic metabolism of indinavir, whether with or without SJW pretreatment. SIGNIFICANCE: Herb-drug pharmacokinetic interactions between SJW and indinavir can be clearly observed in the Wistar rat model. Particularly, the respective first-pass effect contributed by the small intestine and the liver could be differentiated and quantified. The application of the animal model to investigating herb-drug interactions or other relevant research purposes is envisioned. PMID- 19559715 TI - Reproductive numbers, epidemic spread and control in a community of households. AB - Many of the studies on emerging epidemics (such as SARS and pandemic flu) use mass action models to estimate reproductive numbers and the needed control measures. In reality, transmission patterns are more complex due to the presence of various social networks. One level of complexity can be accommodated by considering a community of households. Our study of transmission dynamics in a community of households emphasizes five types of reproductive numbers for the epidemic spread: household-to-household reproductive number, leaky vaccine associated reproductive numbers, perfect vaccine reproductive number, growth rate reproductive number, and the individual reproductive number. Each of those carries different information about the transmission dynamics and the required control measures, and often some of those can be estimated from the data while others cannot. Simulations have shown that under certain scenarios there is an ordering for those reproductive numbers. We have proven a number of ordering inequalities under general assumptions about the individual infectiousness profiles. Those inequalities allow, for instance, to estimate the needed vaccine coverage and other control measures without knowing the various transmission parameters in the models. Along the way, we have also shown that in choosing between increasing vaccine efficacy and increasing coverage levels by the same factor, preference should go to efficacy. PMID- 19559716 TI - Habitat destruction and the extinction debt revisited: the Allee effect. AB - Habitat destruction, often caused by anthropogenic disturbance, can lead to the extinction of species at an unprecedented rate. It is important, therefore, to consider habitat destruction when assessing population viability. Another factor often ignored in population viability analysis, is the Allee effect that adds to the risk of populations already on the verge of extinction. Understanding the Allee effect on species dynamics and response to habitat destruction has intrinsic value in conservation prioritization. Here, the Allee effect was considered in a multi-species hierarchical competition model. Results showed that species persistence declines dramatically due to the Allee effect, and certain species become more susceptible to habitat destruction than others. Two extinction orders emerged under habitat destruction: either the best competitor becomes extinct first or the best colonizer first. The extinction debt and order, as well as the time lag between habitat destruction and species extinction, were found to be determined by species abundance and the intensity of the Allee effect. PMID- 19559717 TI - DNA damage and repair in age-related macular degeneration. AB - Age-related macular degeneration (AMD) is a retinal degenerative disease that is the main cause of vision loss in individuals over the age of 55 in the Western world. Clinically relevant AMD results from damage to the retinal pigment epithelial (RPE) cells thought to be mainly caused by oxidative stress. The stress also affects the DNA of RPE cells, which promotes genome instability in these cells. These effects may coincide with the decrease in the efficacy of DNA repair with age. Therefore individuals with DNA repair impaired more than average for a given age may be more susceptible to AMD if oxidative stress affects their RPE cells. This may be helpful in AMD risk assessment. In the present work we determined the level of basal (measured in the alkaline comet assay) endogenous and endogenous oxidative DNA damage, the susceptibility to exogenous mutagens and the efficacy of DNA repair in lymphocytes of 100 AMD patients and 110 age-matched individuals without visual disturbances. The cells taken from AMD patients displayed a higher extent of basal endogenous DNA damage without differences between patients of dry and wet forms of the disease. DNA double-strand breaks did not contribute to the observed DNA damage as checked by the neutral comet assay and pulsed field gel electrophoresis. The extent of oxidative modification to DNA bases was greater in AMD patients than in the controls, as probed by DNA repair enzymes NTH1 and Fpg. Lymphocytes from AMD patients displayed a higher sensitivity to hydrogen peroxide and UV radiation and repaired lesions induced by these factors less effectively than the cells from the control individuals. We postulate that the impaired efficacy of DNA repair may combine with enhanced sensitivity of RPE cells to blue and UV lights, contributing to the pathogenesis of AMD. PMID- 19559718 TI - Phylogenetic analysis of Bacillus P450 monooxygenases and evaluation of their activity towards steroids. AB - Cytochrome P450 (P450) open reading frames (ORFs) identified in genome sequences of Bacillus species are potential resources for new oxidation biocatalysts. Phylogenetic analysis of 29 Bacillus P450 ORFs revealed that the P450s consist of a limited number of P450 families, CYP102, CYP106, CYP107, CYP109, CYP134, CYP152, and CYP197. Previously, we identified the catalytic activities of three P450s of Bacillus subtilis towards steroids by rapid substrate screening using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). Here, we further applied this method to evaluate the activity of Bacillus cereus P450s towards steroids. Five P450 genes were cloned from B. cereus ATCC 10987 based on its genomic sequence and were expressed in Escherichia coli. These P450s were reacted with a mixture of 30 compounds that mainly included steroids, and the reaction mixtures were analyzed using FT-ICR/MS. We found that BCE_2659 (CYP106) catalyzed the monooxygenation of methyltestosterone, progesterone, 11 ketoprogesterone, medroxyprogesterone acetate, and chlormadinone acetate. BCE_2654 (CYP107) monooxygenated testosterone enanthate, and BCE_3250 (CYP109) monooxygenated testosterone and compactin. Based on the phylogenetic relationship and the known substrate specificities including ones identified in this study, we discuss the catalytic potential of Bacillus P450s towards steroids. PMID- 19559719 TI - Design and synthesis of carbon-11-labeled dual aromatase-steroid sulfatase inhibitors as new potential PET agents for imaging of aromatase and steroid sulfatase expression in breast cancer. AB - Aromatase and steroid sulfatase (STS) are particularly attractive targets in the treatment of estrogen-receptor-positive breast cancer and the development of enzyme-based cancer imaging agents for the biomedical imaging technique positron emission tomography (PET). New carbon-11-labeled sulfamate derivatives were first designed and synthesized as potential PET dual aromatase-steroid sulfatase inhibitor (DASSI) radiotracers for imaging of aromatase and STS expression in breast cancer. The target tracers 5-(((4-cyanophenyl)(4H-1,2,4-triazol-4 yl)amino)methyl)-2-[(11)C]methoxyphenyl sulfamate ([(11)C]8a) and 4-(((4 cyanophenyl)(4H-1,2,4-triazol-4-yl)amino)methyl)-2-[(11)C]methoxyphenyl sulfamate ([(11)C]8b) were prepared from their corresponding precursors 5-(((4 cyanophenyl)(4H-1,2,4-triazol-4-yl)amino)methyl)-2-hydroxyphenyl sulfamate (16) and 4-(((4-cyanophenyl)(4H-1,2,4-triazol-4-yl)amino)methyl)-2-hydroxyphenyl sulfamate (21) with [(11)C]CH(3)OTf under basic conditions through the O [(11)C]methylation and isolated by the reversed-phase high pressure liquid chromatography (HPLC) method in 30-45% radiochemical yields based on [(11)C]CO(2) and decay corrected to end of bombardment (EOB). The specific activity at end of synthesis (EOS) was 111-185GBq/micromol. PMID- 19559720 TI - Quantitative determination of the biological activity of botulinum toxin type A by measuring the compound muscle action potential (CMAP) in rats. AB - Quantitative determination of the biological activity of botulinum toxin type A usually depends on the LD(50) method after intraperitoneal injection into mice. This method requires a large number of mice to determine the toxic activity at a high level of precision and 3-4 days to obtain the results. Techniques to replace the LD(50) method have been attempted at various institutes. As a substitute for this method, by directly measuring the inhibition of neuromuscular transmission after the administration of a toxin, a method to quantitatively assess the toxin's activity by determining the compound muscle action potential (CMAP) was examined. Toxin solutions were injected into the rat gastrocnemius muscle, and that of the CMAP amplitude was determined over time. The CMAP amplitude decreased over 4 days after the injection of the toxin, and then slowly recovered. A dose response relationship was noted for each dose, and a linear relation was observed between 0.01 and 30 U on the 1st day. From these results, we propose the CMAP as a substitute for the LD(50) method to examine the activity of toxin products as it is simple and reliable, reduces the number of experimental animals required, and lowers pain levels. PMID- 19559721 TI - Interaction between visual status, driver age and distracters on daytime driving performance. AB - This study investigated the effects of visual status, driver age and the presence of secondary distracter tasks on driving performance. Twenty young (M=26.8years) and 19 old (M=70.2years) participants drove around a closed-road circuit under three visual (normal, simulated cataracts, blur) and three distracter conditions (none, visual, auditory). Simulated visual impairment, increased driver age and the presence of a distracter task detrimentally affected all measures of driving performance except gap judgments and lane keeping. Significant interaction effects were evident between visual status, age and distracters; simulated cataracts had the most negative impact on performance in the presence of visual distracters and a more negative impact for older drivers. The implications of these findings for driving behaviour and acquisition of driving-related information for people with common visual impairments are discussed. PMID- 19559722 TI - Resveratrol induces senescence-like growth inhibition of U-2 OS cells associated with the instability of telomeric DNA and upregulation of BRCA1. AB - Resveratrol decreases cancer risk and improves health of laboratory animals. However, it can also promote genomic instability. Part of the beneficial activity of resveratrol may result from the activation of SIRT1 deacetylase. We examined how resveratrol influenced the growth of human cancer cell lines of different origin: osteosarcoma (U-2 OS) and lung adenocarcinoma (A549) and how it modulated the expression as well as the localization of key proteins, involved in DNA repair and cell cycle regulation. Resveratrol-induced growth arrest was associated with signs of stress-induced senescence. Differential expression of BRCA1, cyclin B1, pRb and p21 in U-2 OS and A549 cells indicates that resveratrol can engage various molecular mechanisms to arrest cell cycle progression. In subset of U-2 OS cells, the upregulated BRCA1 formed foci closely associated with WRN and the telomeric protein (TRF1). Moreover, resveratrol induced telomeric instability in U-2 OS cells and the activation of DNA damage signaling in both cell lines, manifested as the phosphorylation of histone H2AX at serine 139 and of p53 at serines 15 and 37. Our data are consistent with the hypothesis that resveratrol inhibits cell growth and induces senescence by altering DNA metabolism. PMID- 19559725 TI - In vivo multimodal imaging of stem cell transplantation in a rodent model of Parkinson's disease. AB - Stem cell therapy in the nervous system aims to replace the lost neurons and provide functional recovery. However, it is imperative that we understand the in vivo behaviour of these cells post-implantation. We report visualisation of iron oxide labelled bone marrow-derived stem cells (BMSCs) implanted into the striatum of hemi-parkinsonian rats by magnetic resonance imaging (MRI). Functional efficacy of the donor cells was monitored in vivo using the positron emission tomography (PET) radioligand [11C]raclopride. The cells were visible for 28 days by in vivo MRI. BMSCs provided functional recovery demonstrated by a decreased binding of [11C]raclopride. Although, histology confirmed the persistence of donor cells, no tyrosine hydroxylase positive cells were present. This suggests that BMSCs may have a limited paracrine effect and influence functional recovery. We demonstrate, using multimodal imaging, that we can not only track BMSCs but also establish their effects in a pre-clinical model of Parkinson's disease. PMID- 19559723 TI - Age-related alterations in innate immune receptor expression and ability of macrophages to respond to pathogen challenge in vitro. AB - The impact of ageing in innate immunity is poorly understood. Studies in the mouse model have described altered innate immune functions in aged macrophages, although these were not generally linked to altered expression of receptors or regulatory molecules. Moreover, the influence of ageing in the expression of these molecules has not been systematically examined. We investigated age dependent expression differences in selected Toll-like and other pattern recognition receptors, receptors involved in inflammatory amplification, and in transmembrane and intracellular regulators of inflammatory signaling. Young and aged macrophages were examined under resting conditions or upon activation with Porphyromonas gingivalis, a major pathogen in periodontal disease, the prevalence and severity of which increase in old age. We detected a limited number of age dependent alterations, involving both reduction and increase of immune activity. Interestingly, surface expression of receptors that amplify inflammation (C5a anaphylatoxin receptor and triggering receptor expressed on myeloid cells [TREM] 1) was elevated in aged macrophages. No significant age-dependent differences were observed regarding the phagocytosis and intracellular killing of P. gingivalis, consistent with lack of significant changes in phagocytic receptor expression and induction of antimicrobial molecules. Therefore, at least at the cellular level, certain aspects of innate immune function may not necessarily decline with age. PMID- 19559724 TI - Comparison of the brain penetration injury associated with microdialysis and voltammetry. AB - Emerging evidence suggests that differences between microdialysis- and voltammetry-based estimates of extracellular dopamine in the brain might originate in the different penetration injury associated with each technique. To address this issue in a direct fashion, microdialysis probes and voltammetric microelectrodes were implanted in the rat striatum for 1, 4, or 24 h. Tissues were perfused with a suspension of fluorescently labeled nanobeads to assess blood vessels near the implant. Tissue sections (30 microm) were labeled with antibodies for PECAM, an endothelial cell marker, or GFAP, a glial marker. In non implanted control tissue, blood vessels were reliably double-labeled with nanobeads and antiPECAM. Tissue near microdialysis probe tracks exhibited ischemia in the form of PECAM immunoreactive blood vessels devoid of nanobeads. Ischemia was most apparent after the 4-h implants. Probe tracks were surrounded by endothelial cell debris, which appeared as a diffuse halo of PECAM immunoreactivity. The halo intensity decreased with implant duration, indicative of an active wound-healing process. Consistent with this, after 24-h implants, the probe tracks were surrounded by hyperplasic and hypertrophic glia and glial processes were extending towards, and engulfing, the track. Carbon fiber microelectrodes produced a diffuse disruption of nanobead labeling but no focal disruption of blood vessels, no PECAM immunoreactive halo, and no glial activation. These findings illuminate the differences between the extent and nature of the penetration injuries associated with microdialysis and voltammetry. PMID- 19559726 TI - Exogenous HIV-1 Vpr disrupts IFN-alpha response by plasmacytoid dendritic cells (pDCs) and subsequent pDC/NK interplay. AB - HIV Vpr is known for its immunomodulatory capacities including its impairment of NK cell functions. However, the role of pDCs in this context remains elusive. We show that synthetic Vpr substantially inhibits type I IFN production by pDCs without inducing apoptosis in pDCs. Furthermore, we found that exogenous Vpr compromises subsequent pDC/NK interplay as shown by diminished IFN-gamma production by NK cells. Thus, Vpr-mediated dysregulation of IFN-alpha and IFN gamma production affects key components of the innate immune response supporting an essential role of Vpr in HIV pathogenesis. PMID- 19559727 TI - Isolation and characterization of human neutralizing antibodies to rabies virus derived from a recombinant immune antibody library. AB - A human immune Fab library was constructed using RNAs from peripheral blood lymphocytes obtained from rabies virus hyperimmune volunteers on phagemid vector. The size of the constructed Fab library was 2 x 10(7) Escherichia coli transformants. After four rounds of panning on whole inactivated rabies virus (PV 11), phage clones displaying rabies virus-specific human Fab were selected. The specificity of soluble Fab antibody fragments, derived from positive phage clones was verified by ELISA. Among 20 specific Fab clones, the genetic sequence of 6 of them (FabRV01, FabRV02, FabRV03, FabRV04, FabRV05, and FabRV06) was analyzed. The variable heavy (VH) and variable light (VL) domains were found to share 90% and 93% homology with sequences encoded by the corresponding human germline genes, respectively. The soluble Fab fragments, expressed in Escherichia coli were purified by a single step Nickel-NTA affinity chromatography via a hexa-histidine tag and their binding specificities to rabies virus were confirmed. Three of the Fab antibodies, FabRV01, FabRV02 and FabRV03, showed binding characteristics to rabies virus glycoprotein antigenic site III with affinities in the K(D) range 7 x 10(-9) to 5 x 10(-8)M. The Fab fragments showed dose-dependent neutralization properties for the challenge virus standard (CVS-11). PMID- 19559728 TI - Detection and identification of U69 gene mutations encoded by ganciclovir resistant human herpesvirus 6 using denaturing high-performance liquid chromatography. AB - A denaturing high-performance liquid chromatography (dHPLC) assay was developed to detect antiviral drug-resistance mutations of human herpesvirus 6 (HHV-6). Recombinant baculoviruses were created that contained wild-type and mutant forms of the HHV-6 U69 gene, which determines sensitivity to the antiviral drug ganciclovir (GCV). The mutations causing GCV resistance in HHV-6 U69 were single base mutations adapted from known GCV-resistant DNA sequences of HCMV, and their ability to confer GCV resistance on recombinant baculoviruses was confirmed. Six characterized mutant sequences, including one reported previously that encodes a GCV-sensitive kinase-activity mutant, were used. DNA was extracted, and the levels of homoduplex and heteroduplex DNA in the PCR products from mixed wild type and mutant viral DNAs were determined using dHPLC. The optimized assay could distinguish the different mutants, and could detect mutants representing only 10% of the DNAs. The new assay with dHPLC readout permitted the rapid (4 h), objective, and reproducible detection of HHV-6 drug-resistance mutations. PMID- 19559729 TI - One-step real-time quantitative PCR assays for the detection and field study of Sacbrood honeybee and Acute bee paralysis viruses. AB - Two one-step real-time RT-PCR assays, based on SYBR Green (SG) chemistry, were developed or adapted respectively, for the detection, differentiation, and quantitation of two important honeybee viruses: Sacbrood virus (SBV) and Acute bee paralysis virus (ABPV). Both reactions were optimized to yield the highest sensitivity and specificity. The genome equivalent copies (GEC) detection limit per reaction was 389.3 for the ABPV RT-PCR. The GEC detection limit per reaction was 298.9 for the SBV RT-PCR. Viral detection and identification were confirmed by melting curve analysis and sequencing of the PCR products. Both techniques were used to evaluate Spanish field samples and establish the distribution of these viruses. Acute bee paralysis virus was not detected, and Sacbrood virus was present at low frequencies. The one-step real-time SG RT-PCR methods are fast, accurate, and useful for detecting and quantifying these honeybee viruses, which cause inapparent infections and contribute to the increasing depopulation of honeybee colonies. PMID- 19559730 TI - A combined genotypic and phenotypic human immunodeficiency virus type 1 recombinant virus assay for the reverse transcriptase and integrase genes. AB - With the approval of the first HIV-1 integrase inhibitor raltegravir and a second one in phase III clinical development (elvitegravir), genotypic and phenotypic resistance assays are required to guide antiretroviral therapy and to investigate treatment failure. In this study, a genotypic and phenotypic recombinant virus assay was validated for determining resistance against integrase inhibitors. The assays are based on the amplification of a region encompassing not only HIV-1 integrase, but also reverse transcriptase and RNAseH. The overall amplification success was 85% (433/513) and increased to 93% (120/129) for samples with a viral load above 3 log(10) copies/ml. Both B and non-B HIV-1 subtypes could be genotyped successfully (93%; 52/56 and 100%; 49/49, respectively) and reproducibly. The phenotypic assay showed a high success rate (96.5%; 139/144) for subtype B (100%; 19/19) and non-B subtypes (92%; 45/49), and was found to be accurate and reproducible as assessed using well-characterized integrase mutants. Using both assays, baseline resistance to raltegravir and elvitegravir in subtype B and non-B HIV-1 strains selected at random was not observed, although integrase polymorphisms were present at varying prevalence. Biological cutoff values were found to be 2.1 and 2.0 for raltegravir and elvitegravir, respectively. In summary, a genotypic and phenotypic integrase resistance assay was validated successfully for accuracy, reproducibility, analytical and clinical sensitivity, and dynamic range. PMID- 19559731 TI - Monitoring of HIV-1 envelope-mediated membrane fusion using modified split green fluorescent proteins. AB - A simple, cell-based, membrane fusion assay system that uses split green fluorescent proteins (spGFPs) as an indicator was developed. The attachment of the pleckstrin homology (PH) domain to the N-termini of each spGFP not only localized the reporter signal to the plasma membrane but also helped the stable expression of the smaller spGFP of seventeen amino acid residues. It was shown that this system allowed real-time monitoring of membrane fusion by HIV-1 envelope protein (Env) without the addition of external substrates. This method can be adapted to the analyses of other viral membrane fusion. PMID- 19559732 TI - CD4-BFFI: a novel, bifunctional HIV-1 entry inhibitor with high and broad antiviral potency. AB - Resistance to antiretroviral drugs is a common problem in the treatment of HIV-1 infected patients. To overcome resistance, we generated a novel, bifunctional HIV 1 entry inhibitor by combining the anti-CD4 monoclonal antibody (mAb) 6314 with a fusion inhibitor similar to T-651 (anti-CD4 mAb based BiFunctional Fusion Inhibitor, CD4-BFFI). CD4-BFFI has potent antiviral activity against a multitude of HIV-1 isolates independent of their co-receptor usage and genetic background. It has higher antiviral potency compared to the fusion inhibitor T-651 or the anti-CD4 mAb 6314 used independently. More importantly, every HIV-1 strain tested was fully inhibited by CD4-BFFI while many strains were only partially inhibited by 6314. CD4-BFFI also retained antiviral potency against virus strains resistant to two fusion inhibitors, a CCR5 antagonist and an anti-CCR5 mAb. Pre-incubation of cells with a saturating concentration of anti-CD4 mAbs reduced the antiviral potency of CD4-BFFI, suggesting that binding of CD4-BFFI to the cell surface via its CD4 mAb portion is required for the antiviral potency of its fusion inhibitor moiety. Collectively, we present a novel HIV-1 inhibitor with a dual mode of action and excellent antiviral potency against wildtype and entry-inhibitor resistant virus strains suggesting that CD4-BFFI may have a high barrier to resistance. PMID- 19559734 TI - Identification and quantification of sophorolipid analogs using ultra-fast liquid chromatography-mass spectrometry. AB - An ultra-fast liquid chromatographic method combined with atmospheric pressure chemical ionization mass detection (UHPLC/APCI-MS) has been developed for the separation and quantification of sophorolipid analogs produced by the yeast Candida bombicola. The sophorolipid mixture was produced by growing the yeast in the presence of glucose and oleic acid under higher aeration. It was found that more than 95% of the analogs are lactonic sophorolipids and all the produced sophorolipids produced were either mono- or di-acetylated. Also observed was a sophorolipid analog with a tri-unsaturated fatty acid, which has not been reported previously. PMID- 19559733 TI - Analysis of expressed sequence tags from the four main developmental stages of Trypanosoma congolense. AB - Trypanosoma congolense is one of the most economically important pathogens of livestock in Africa. Culture-derived parasites of each of the three main insect stages of the T. congolense life cycle, i.e., the procyclic, epimastigote and metacyclic stages, and bloodstream stage parasites isolated from infected mice, were used to construct stage-specific cDNA libraries and expressed sequence tags (ESTs or cDNA clones) in each library were sequenced. Thirteen EST clusters encoding different variant surface glycoproteins (VSGs) were detected in the metacyclic library and 26 VSG EST clusters were found in the bloodstream library, 6 of which are shared by the metacyclic library. Rare VSG ESTs are present in the epimastigote library, and none were detected in the procyclic library. ESTs encoding enzymes that catalyze oxidative phosphorylation and amino acid metabolism are about twice as abundant in the procyclic and epimastigote stages as in the metacyclic and bloodstream stages. In contrast, ESTs encoding enzymes involved in glycolysis, the citric acid cycle and nucleotide metabolism are about the same in all four developmental stages. Cysteine proteases, kinases and phosphatases are the most abundant enzyme groups represented by the ESTs. All four libraries contain T. congolense-specific expressed sequences not present in the Trypanosoma brucei and Trypanosoma cruzi genomes. Normalized cDNA libraries were constructed from the metacyclic and bloodstream stages, and found to be further enriched for T. congolense-specific ESTs. Given that cultured T. congolense offers an experimental advantage over other African trypanosome species, these ESTs provide a basis for further investigation of the molecular properties of these four developmental stages, especially the epimastigote and metacyclic stages for which it is difficult to obtain large quantities of organisms. The T. congolense EST databases are available at: http://www.sanger.ac.uk/Projects/T_congolense/EST_index.shtml. The sequence data have been submitted to EMBL under the following accession numbers: FN263376 FN292969. PMID- 19559735 TI - A comparison of fluorescent DNA binding dyes for flow cytometric analysis of sporulating Saccharomyces cerevisiae. AB - Flow cytometry is important tool for investigating DNA replication in sporulating Saccharomyces cerevisiae. However, flow cytometry data from maturing spores is often difficult to interpret due to extensive broadening of the fluorescence peaks. This problem is markedly improved by treatment of the spores with potassium hydroxide prior to staining. PMID- 19559736 TI - Physiological levels of HBB transgene expression from S/MAR element-based replicating episomal vectors. AB - Replicating episomal vectors (REV) are in principle able to provide long-term transgene expression in the absence of integration into the target cell genome. The scaffold/matrix attachment region (S/MAR) located 5' of the human beta interferon gene (IFNB1) has been shown to confer a stable episomal replication and retention function within plasmid vectors when stably transfected and selected in mammalian cells. The minimal requirement for the IFNB1 S/MAR to function in DNA replication and episomal retention is transcription through this element. We used the erythroid beta-globin locus control region-beta-globin gene (betaLCR-HBB) microlocus cassette as a model to assess tissue-specific expression from within an IFNB1 S/MAR-based plasmid REV. The betaLCR-HBB plus S/MAR combination constructs provided either high or low levels of transcription through the S/MAR element. Our results show that the betaLCR-HBB microlocus is able to reproducibly and stably express at full physiological levels on an episome copy number basis. In addition, our data show that even low levels of transcription from betaLCR-HBB through the S/MAR element are sufficient to allow efficient episomal replication and retention. These data provide the principles upon which generic and flexible expression cassette-S/MAR-based REVs can be designed for a wide range of applications. PMID- 19559737 TI - Physiological regulation of laccase and manganese peroxidase production by white rot Basidiomycetes. AB - This review integrates recent literature and our own data on the physiology of laccase and manganese peroxidase synthesis, focusing on the common characteristics and unique properties of individual fungi as well as on several approaches providing enhanced enzyme secretion. Firstly, the enzyme yield is species-dependent and strain-dependent and selection of new organisms with tremendous synthesis of these enzymes is possible. For example, in screening program the laccase activity of tested basidiomycetes varied from 0.5Uml(-1) to 75Uml(-1). Secondly, the carbon source and lignocellulosic substrate play a crucial role in enzyme production. Thus, laccase activity of Pseudotrametes gibbosa varied from 0.3Uml(-1) (Avicel) to 13.7Uml(-1) (lactose), while the substitution of wheat bran with walnut pericarp increased Cerrena unicolor manganese peroxidase yield from 0.7Uml(-1) to 8.3Uml(-1). Thirdly, aromatic compounds regulate the ligninolytic enzyme synthesis although their effect is very specific depending on fungi physiological peculiarities. 2,4,6 trinitrotoluene (TNT) supplemented to the medium at appropriate concentration significantly accelerated C. unicolor laccase production and 4-fold increased laccase specific activity. Fourthly, co-cultivation of appropriate fungi shows considerable promise as a strategy to highly enhance the enzyme production. For example, pairing of C. unicolor and Phellinus robustus 2-fold increased the total laccase yield. PMID- 19559738 TI - Generation of Tioman virus nucleocapsid-like particles in yeast Saccharomyces cerevisiae. AB - Tioman virus (TioV) was isolated from a number of pooled urine samples of Tioman Island flying foxes (Pteropus hypomelanus) during the search for the reservoir host of Nipah virus. Studies have established TioV as a new virus in the family Paramyxoviridae. This novel paramyxovirus is antigenically related to Menangle virus that was isolated in Australia in 1997 during disease outbreak in pigs. TioV causes mild disease in pigs and has a predilection for lymphoid tissues. Recent serosurvey showed that 1.8% of Tioman Islanders had neutralizing antibodies against TioV, indicating probable past infection. For the development of convenient serological tests for this virus, recombinant TioV nucleocapsid (N) protein was expressed in the yeast Saccharomyces cerevisiae. High yields of recombinant TioV N protein were obtained. Electron microscopy demonstrated that purified recombinant N protein self-assembled into nucleocapsid-like particles which were identical in density and morphology to authentic nucleocapsids from paramyxovirus-infected cells. Different size nucleocapsid-like particles were stable and readily purified by CsCl gradient ultracentrifugation. Polyclonal sera raised in rabbits after immunization with recombinant TioV N protein reacted reliably with TioV infected tissues in immunohistochemistry tests. It confirmed that the antigenic properties of yeast derived TioV N protein are identical to authentic viral protein. PMID- 19559740 TI - Oral strip technology: overview and future potential. AB - Over the recent past, many of the research groups are focusing their research on this technology. Amongst the plethora of avenues explored for rapid drug releasing products, Oral Strip Technology (OST) is gaining much attention. The advantages of OST are the administration to pediatric and geriatric patient population where the difficulty of swallowing larger oral dosage forms is eliminated. This technology has been used for local action, rapid release products and for buccoadhesive systems that are retained for longer period in the oral cavity to release drug in controlled fashion. OST offers an alternate platform for molecules that undergo first pass metabolism and for delivery of peptides. The review article is an overview of OST encompassing materials used in OST, critical manufacturing aspects, applications, commercial technologies and future business prospects of this technology. PMID- 19559741 TI - Confocal Raman spectroscopy to study in vivo skin penetration of retinol. PMID- 19559739 TI - Molecular variation analysis of porcine reproductive and respiratory syndrome virus in China. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) is characteristic of genetically extensive variation. The objective of the present study was to analyze the molecular variation and evolution of porcine reproductive and respiratory syndrome virus in China based on the complete genomic sequences of three highly pathogenic Chinese PRRSV strains isolated in 2006 and the sequences of the amplified Nsp2, ORF5 and ORF7 genes from clinical specimens during 2006 2008. Full-length genome sequencing and phylogenetic analysis showed that the three strains (JXwn06, BJsy06 and NX06) had a unique 30-amino-acid discontinuous deletion in Nsp2, and were classified into the same subgroup that consisted of the most Chinese strains isolated during 2006-2007, the pandemic period of atypical PRRS. The evolution analysis suggested that the emergence of the highly pathogenic PRRSV in China experienced a gradual variation and evolution accumulation progress from Chinese domestic virus. The variation analysis of the amplified 41 Nsp2, 59 ORF5 and 59 ORF7 genes indicated that the diversity of PRRSV strain existed in the field, and the highly pathogenic PRRSV strain with the 30-amino-acid deletion in Nsp2 was the dominating virus in China in recent years. Our data contribute to the understanding of molecular variation and epidemiology surveillance of PRRSV in China. PMID- 19559742 TI - Oral tolerance: can we make it work? AB - Mucosal tolerance remains an attractive approach for the treatment of autoimmune and inflammatory diseases. The agents used in these treatments lack toxicity, can be easily administered, and enable the promotion of antigen-specific immune responses. The limited success of clinical trials over the past 2 decades has led to the fear that the beneficial effect observed in animal models cannot be repeated in humans. Successful application of mucosal tolerance for the treatment of human diseases will depend on strategies that target the correct cells in the gut-liver axis, improve antigen presentation, alter the administered dose and formulations, utilize potent mucosal adjuvants, develop immune biomarkers enabling follow-up of the effect, utilize combination therapies with other immune modulatory agents, and target the right patient populations. Here, we discuss 12 of the major questions related to oral tolerance and its clinical application to humans with immune-mediated disorders. PMID- 19559743 TI - Comparison of degranulation of easily mobilizable intracellular granules by human phagocytes in healthy subjects and patients with infectious diseases. AB - The aim of this study was to compare degranulation of easily mobilizable secretory vesicles (SVs) or secretory vesicle-like granules (SVLGs) in neutrophils, monocytes, and eosinophils of healthy controls (n = 60) and febrile patients with microbiologically confirmed or clinically diagnosed bacterial (n = 89) and viral (n = 46) infections. For this purpose, flow cytometric immunophenotyping of isolated phagocytes was performed using monoclonal antibodies against the phagocytosis receptors CR1 (CD35) and CR3 (CD11b) that are predominantly stored in the SVs of resting neutrophils. Similar to neutrophils, monocytes contain easily mobilizable SVLGs that constitute the main intracellular reservoir of CD35 and CD11b. In both neutrophils and monocytes, activation mechanisms leading to degranulation of SV and SVLG appeared dependent on both intra- and extracellular calcium levels. The kinetics of degranulation of SVLGs in control monocytes was significantly faster than that of SVs of control neutrophils. We conclude that phagocytes in patients with bacterial infections can be arranged in order of decreasing magnitude of SV or SVLG degranulation as follows (from left to right): neutrophils > monocytes " eosinophils. However, in viral infections, the corresponding degranulation order is monocytes > neutrophils approximately eosinophils. PMID- 19559744 TI - 49A/G and CT60 polymorphisms of the cytotoxic T-lymphocyte-associated antigen 4 gene associated with autoimmune thyroid disease. AB - Single nucleotide polymorphisms in the CTLA-4 gene have been suggested as genetic factors in the susceptibility to autoimmune thyroid disease (AITD). In our case control study, patients with Graves' disease, Hashimoto's thyroiditis, and postpartum thyroiditis and control subjects have been genotyped for two A/G single nucleotide polymorphisms (49A/G and CT60) of the CTLA-4 gene. The 49A/G polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism method using the enzyme BseXI and the CT60 polymorphism by real-time polymerase chain reaction. Results were analyzed by chi(2) test and linkage disequilibrium analysis. In a comparison of frequencies of GG genotype, a significant association of 49A/G and CT60 polymorphism existed only for Graves' disease. In the 49A/G polymorphism, the frequency of GG genotype was significantly higher (p = 0.0408) compared with controls; the frequency of the CT60 polymorphism was significantly higher as well (p = 0.0213). The frequencies of AA and AG genotypes in control subjects did not significantly differ from frequencies in AITD patients for both polymorphisms. Our results may therefore lend support to the hypothesis that humoral autoimmunity is correlated with 49A/G and CT60 polymorphisms of the CTLA-4 gene. PMID- 19559745 TI - A sub-chronic (13 weeks) oral toxicity study in rats and an in vitro genotoxicity study with Korean pine nut oil (PinnoThin TG). AB - In this paper a sub-chronic (13 weeks) toxicity study in rats and an in vitro genotoxicity study with Korean pine (Pinus koraiensis Siebold & Zucc.) nut oil, KPNO (PinnoThin) are described. Both studies were performed in compliance with GLP, and in line with OECD guidelines applicable. In the sub-chronic toxicity study, no clinical signs, abnormalities in functional observation tests or ophthalmologic examinations or changes in body weight or food intake were noted at any of the doses of KPNO tested. Various changes in clinical biochemistry parameters were noted. Whilst these changes were not consistent in both sexes, and neither associated with any histopathological changes, nor dose-related, these were not considered to be toxicologically relevant. No toxicologically significant changes were noted in haematological parameters. There were a few histopathological observations such as a periportal vacuolation of the liver in all dose groups including the control, and renal tubular mineralisation in most females of the high dose group but also in all control female rats. These findings can be considered to be due to the high fat content of the diets, and are not related to the treatment with KPNO. Based on these findings a No Observable Adverse Effect Level (NOAEL) of 15% has been established for KPNO. This NOAEL corresponded to a mean of 8866 and 10,242 mg KPNO/kg bw/day for males and females, respectively. This dose level was the highest achievable oral dose for KPNO in rats. The in vitro reverse mutation test (Ames test), showed no significant dose-related increase in the number of revertants in two independently repeated mutation assays. The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly. Based on these results it has been concluded that KPNO is not mutagenic in the Escherichia coli and Salmonella typhimurium reverse mutation assays. In conclusion, KPNO can be considered to be non-genotoxic in the AMES test. A NOAEL of 8866 and 10,242 mg KPNO/kg bw/day has been established for male and female rats, respectively. For both sexes, the NOAEL was achieved at the highest dose tested. PMID- 19559746 TI - Tri-n-butyltin increases intracellular Zn(2+) concentration by decreasing cellular thiol content in rat thymocytes. AB - Effect of tri-n-butyltin (TBT), an environmental pollutant, on intracellular Zn(2+) concentration was tested in rat thymocytes to reveal one of cytotoxic profiles of TBT at nanomolar concentrations using a flow cytometer and appropriate fluorescent probes. TBT at concentrations of 30 nM or more (up to 300 nM) significantly increased the intensity of FluoZin-3 fluorescence, an indicator for intracellular Zn(2+) concentration, under external Ca(2+)- and Zn(2+)-free condition. Chelating intracellular Zn(2+) completely attenuated the TBT-induced augmentation of FluoZin-3 fluorescence. Result suggests that nanomolar TBT releases Zn(2+) from intracellular store site. Oxidative stress induced by hydrogen peroxide also increased the FluoZin-3 fluorescence intensity. The effects of TBT and hydrogen peroxide on the fluorescence were additive. TBT induced changes in the fluorescence of FluoZin-3 and 5-chloromethylfluorescein, an indicator for cellular thiol content, were correlated with a coefficient of 0.962. Result suggests that the intracellular Zn(2+) release by TBT is associated with TBT-induced reduction of cellular thiol content. However, chelating intracellular Zn(2+) potentiated the cytotoxicity of TBT. Therefore, the TBT induced increase in intracellular Zn(2+) concentration may be a type of stress responses to protect the cells. PMID- 19559747 TI - Deep brain stimulation in neurological diseases and experimental models: from molecule to complex behavior. AB - Deep brain stimulation (DBS) has proven to be capable of providing significant benefits for several neuropathologies. It is highly effective in reducing the motor symptoms of Parkinson's disease, essential tremor, and dystonia, and in alleviating chronic pain. Recently, also Tourette syndrome, obsessive-compulsive disorder and treatment-resistant depression have been treated by DBS with encouraging results. However, despite these clinical achievements, the precise action mechanisms of DBS still need to be fully characterized. For this reason, several animal models of DBS have been developed, bringing new insights on the effects of this treatment at molecular and cellular level, and providing new evidence on its physiological and behavioral consequences. In parallel, physiological and imaging studies in patients have contributed to better understanding DBS impact on the function of brain circuits. Here we review the clinical data and experimental work in vitro, ex vivo and in vivo (mostly arisen from studies on DBS of the subthalamic nucleus) in the treatment of PD, which led to the actual knowledge of DBS mechanisms, from molecular to complex behavioral levels. PMID- 19559749 TI - Simple scaling of cooperation in donor-recipient games. AB - We present a simple argument which proves a general version of the scaling phenomenon recently observed in donor-recipient games by Tanimoto [Tanimoto, J., 2009. A simple scaling of the effectiveness of supporting mutual cooperation in donor-recipient games by various reciprocity mechanisms. BioSystems 96, 29-34]. PMID- 19559748 TI - Breathing frequency bias in fractal analysis of heart rate variability. AB - Detrended Fluctuation Analysis (DFA) is an algorithm widely used to determine fractal long-range correlations in physiological signals. Its application to heart rate variability (HRV) has proven useful in distinguishing healthy subjects from patients with cardiovascular disease. In this study we examined the effect of respiratory sinus arrhythmia (RSA) on the performance of DFA applied to HRV. Predictions based on a mathematical model were compared with those obtained from a sample of 14 normal subjects at three breathing frequencies: 0.1Hz, 0.2Hz and 0.25Hz. Results revealed that: (1) the periodical properties of RSA produce a change of the correlation exponent in HRV at a scale corresponding to the respiratory period, (2) the short-term DFA exponent is significantly reduced when breathing frequency rises from 0.1Hz to 0.2Hz. These findings raise important methodological questions regarding the application of fractal measures to short term HRV. PMID- 19559750 TI - Regulation of preprosomatostatin 1 (PSS1) gene expression by 17beta-estradiol and identification of the PSS1 promoter region in orange-spotted grouper (Epinephelus coioides). AB - In the present paper the effects of 17beta-estradiol on the expression of the preprosomatostatin 1 (PSS1) in the orange-spotted grouper hypothalamus and ovary were investigated. Results from in vivo of intraperitoneal injection and in vitro static cultures showed that estradiol increased the mRNA expression of PSS1 gene in both hypothalamus and ovary. To investigate the molecular basis of the estrogen regulation on PSS1 gene expression, we cloned the upstream region of 848bp from the translation initiation codon of the grouper PSS1 gene. The TATA box and putative transcription factor binding sites were identified using computer analysis. Transient transfections with promoter-luciferase reporter constructs together with hER expression vector were carried out in MCF-7 cell line. The results suggest that the region from -848 to -373bp, containing five putative ERE half sites, may contribute to the promoter activity induced by estradiol. These results represent the first demonstration at the molecular level of the regulation of PSS1 gene by 17beta-estradiol in fish. PMID- 19559751 TI - Removing ECG noise from surface EMG signals using adaptive filtering. AB - Surface electromyograms (EMGs) are valuable in the pathophysiological study and clinical treatment for dystonia. These recordings are critically often contaminated by cardiac artefact. Our objective of this study was to evaluate the performance of an adaptive noise cancellation filter in removing electrocardiogram (ECG) interference from surface EMGs recorded from the trapezius muscles of patients with cervical dystonia. Performance of the proposed recursive-least-square adaptive filter was first quantified by coherence and signal-to-noise ratio measures in simulated noisy EMG signals. The influence of parameters such as the signal-to-noise ratio, forgetting factor, filter order and regularization factor were assessed. Fast convergence of the recursive-least square algorithm enabled the filter to track complex dystonic EMGs and effectively remove ECG noise. This adaptive filter procedure proved a reliable and efficient tool to remove ECG artefact from surface EMGs with mixed and varied patterns of transient, short and long lasting dystonic contractions. PMID- 19559753 TI - Functional analysis of novel KCNQ2 mutations found in patients with Benign Familial Neonatal Convulsions. AB - Benign Familial Neonatal Convulsions (BFNC) are a rare epilepsy disorder with an autosomal-dominant inheritance. It is linked to mutations in the potassium channel genes KCNQ2 and KCNQ3. These encode for Kv7.2 and Kv7.3 potassium ion channels, which produce an M-current that regulates the potential firing action in neurons through modulation of the membrane potential. We report on the biophysical and biochemical properties of V589X, T359K and P410fs12X mutant-KCNQ2 ion channels that were detected in three BFNC families. Mutant KCNQ2 cDNAs were co-expressed with WT-KCNQ2 and KCNQ3 cDNAs in HEK293 cells to mimic heterozygous expression of the KCNQ2 mutations in BFNC patients. The resulting potassium currents were measured using patch-clamp techniques and showed an approximately 75% reduction in current and a depolarized shift in the voltage dependence of activation. Furthermore, the time-constant of activation of M-currents in cells expressing T359K and P410fs12X was slower compared to cells expressing only wild type proteins. Immunofluorescent labeling of HEK293 cells stably expressing GFP tagged KCNQ2-WT or mutant alpha-subunits indicated cell surface expression of WT, V589X and T359K mutants, suggesting a loss-of-function, while P410fs12X was predominantly retained in the ER and sub-cellular compartments outside the ER suggesting an effectively haplo-insufficient effect. PMID- 19559752 TI - Rotenone and paraquat do not directly activate microglia or induce inflammatory cytokine release. AB - Both epidemiological and pathological data suggest an inflammatory response including microglia activation and neuro-inflammation in the Parkinsonian brain. Treatments with lipopolysaccharide (LPS), rotenone and paraquat have been used as models for Parkinson's disease, as they cause dopaminergic neuron degeneration in culture and in animals. Recent studies have suggested that rotenone and paraquat induce neuro-inflammation, however, it is not known if they can directly activate microglia. Here, we use primary cultured microglia to address this question. Microglia activation was analyzed by morphological changes and release of nitric oxide and inflammatory cytokines. Treatment with LPS was used as a positive control. While LPS induced morphological changes characteristic of microglial activation and release of nitric oxide and inflammatory cytokines, rotenone and paraquat did not. Our results suggest that paraquat and rotenone do not act directly on microglia and that neuro-inflammation and microglial activation in animals treated with these agents are likely non-cell autonomous, and may occur as a result of dopaminergic neuron damage or factors released by neurons and other cells. PMID- 19559754 TI - Difference of climbing fiber input sources between the primate oculomotor-related cerebellar vermis and hemisphere revealed by a retrograde tracing study. AB - The cerebellar flocculus-paraflocculus complex, vermal lobule VII (V-7) and hemispheric lobule VII (H-7) are involved in learning-dependent smooth pursuit eye movement control. To locate the sources of climbing fiber inputs to the H-7 and V-7, we injected retrograde tracers and examined the locations of retrogradely labeled neurons in the inferior olive in 4 monkeys. After the injection of cholera toxin B (CTB) into the H-7, retrogradely labeled neurons were observed abundantly in cell group d, i.e., dorsal cap, of the caudal medial accessory olive (MAO) and ventral lamella of principal olive (PO). After injections of fast blue (FB) into the V-7, retrogradely labeled neurons were observed mainly in cell group b of MAO, but rarely in cell group d or PO. Cell group d is known to receive inputs from the nucleus optic tract (NOT) and project climbing fibers to the flocculus and ventral paraflocculus, and cell group b is known to receive inputs from the superior colliculus. These results suggest that the three oculomotor cerebellar areas may use different visual signals for the control of smooth pursuit: the flocculus-paraflocculus complex and H-7 receive visual climbing fiber inputs derived mainly from the NOT via cell group d, while the V-7 receive visual climbing fiber inputs derived mainly from the superior colliculus via cell group b. PMID- 19559756 TI - Estrogens, MSI and Lynch syndrome-associated tumors. AB - Estrogens play a major role in the biology of hormone-responsive tissues but also in the normal physiology of various non-typical hormone-responsive tissues. In disease, estrogens have been associated with tumor development, in particular with tumors such as breast, endometrium, ovary and prostate. In this paper we will review the molecular mechanisms by which estrogens are involved in cancer development, with a special focus in Lynch syndrome related neoplasia. Further, we discuss the role estrogens might have on cell proliferation and apoptosis, how estrogens metabolites can induce DNA damage and we discuss a possible connection between estrogens and changes in DNA (hypo- and hyper) methylation. In this review we will also address the protective effect that high levels of estrogens have in MMR related neoplasias. PMID- 19559758 TI - New insights in cerebellar function. Introduction. PMID- 19559757 TI - Glutamate release upon purinergic action in the dorsal facial area of the medulla increases blood flow in the common carotid artery in cats. AB - Interactions of glutamatergic and purinergic actions in the medulla regulate important cardiovascular functions. The glutamatergic action in dorsal facial area (DFA) of the medulla increases blood flow of common carotid artery (CCA) in cats. We hypothesized that interactions of glutamatergic and purinergic actions in the DFA may regulate the CCA blood flow. Purinergic and glutamatergic agonists and antagonists were microinjected into the DFA through a four-barrel tubing in anesthetized cats. Drug effects were evaluated by changes in the CCA blood flow. Microinjection with 20 nmol ATP or alpha,beta-methyleneATP (alpha,beta-MeATP, a P2 purinergic receptor agonist) induced an increase of the CCA blood flow. This increase was dose-dependently reduced by prior administration with 1,3-dipropyl-8 p-sulfophenylxanthine (DPSPX, a specific P1 purinergic receptor antagonist), or pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS, a selective P2 purinergic receptor antagonist) as well as with MK-801 (a non-competitive NMDA receptor antagonist) or glutamate diethyl ester (GDEE, a competitive AMPA/kainate receptor antagonist). It was almost completely blocked by administrations with combined maximal doses of P1 and P2 receptor antagonists as well as NMDA and AMPA receptor antagonists. Nevertheless, P1 receptor agonist induced only mild and poorly reproducible increase in the CCA blood flow. In conclusion, prominent P2 and minor P1 purinergic receptors appear to be present in the DFA; the purinergic activation can mediate a release of glutamate that stimulates NMDA and AMPA to induce the increase of the CCA blood flows. These findings may provide important information for developing therapeutic strategy for diseases involving the CCA blood flow, such as hypertensive disease and cerebral ischemia. PMID- 19559759 TI - Genetics of emotion regulation. AB - Emotions can be powerful drivers of behavior that may be adaptive or maladaptive for the individual. Thus, the ability to alter one's emotions, to regulate them, should be beneficial to an individual's success of survival and fitness. What is the biological basis of this ability? And what are the biological mechanisms that impart individual differences in the ability to regulate emotion? In this article, we will first introduce readers to the construct of emotion regulation, and the various strategies that individuals may utilize to regulate their emotions. We will then point to evidence that suggests genetic contributions (alongside environmental contributions) to individual differences in emotion regulation. To date, efforts to identify specific genetic mechanisms involved in emotion regulation have focused on common gene variants (i.e. variants that exist in >1% of the population, referred to as polymorphisms) and their association with specific emotion regulation strategies or the neural substrate mediating these strategies. We will discuss these efforts, and conclude with a call to expand the set of experimental paradigms and putative molecular mechanisms, in order to significantly advance our understanding of the molecular mechanisms by which genes are involved in emotion regulation. PMID- 19559755 TI - Epigenetic mechanisms in schizophrenia. AB - Epidemiological research suggests that both an individual's genes and the environment underlie the pathophysiology of schizophrenia. Molecular mechanisms mediating the interplay between genes and the environment are likely to have a significant role in the onset of the disorder. Recent work indicates that epigenetic mechanisms, or the chemical markings of the DNA and the surrounding histone proteins, remain labile through the lifespan and can be altered by environmental factors. Thus, epigenetic mechanisms are an attractive molecular hypothesis for environmental contributions to schizophrenia. In this review, we first present an overview of schizophrenia and discuss the role of nature versus nurture in its pathology, where 'nature' is considered to be inherited or genetic vulnerability to schizophrenia, and 'nurture' is proposed to exert its effects through epigenetic mechanisms. Second, we define DNA methylation and discuss the evidence for its role in schizophrenia. Third, we define posttranslational histone modifications and discuss their place in schizophrenia. This research is likely to lead to the development of epigenetic therapy, which holds the promise of alleviating cognitive deficits associated with schizophrenia. PMID- 19559760 TI - Reduced expression and desensitization of adenosine A1 receptor/adenylyl cyclase pathway after chronic (-)N6-phenylisopropyladenosine intake during pregnancy. AB - Little is known about the G protein-coupled receptor desensitization process during pregnancy. Wistar pregnant rats were treated with (-)N(6) phenylisopropyladenosine (R-PIA), an adenosine A(1) receptor (A(1)R) agonist, in their drinking water during pregnancy, and the effect on A(1)R/adenylyl cyclase system was studied in both maternal and fetal brain. In maternal brain, binding assays revealed a significant decrease in total receptor numbers in plasma membranes (27%, P<0.05), with no significant changes in receptor affinity. The effect of R-PIA on plasma membranes from fetal brains was more marked, with approximately 42% (P<0.05) of the total receptors detected in control fetuses. Real time reverse transcriptase polymerase chain reaction (RT-PCR) analyses showed that chronic R-PIA treatment during the whole gestational period only decreased significantly mRNA level coding A(1)R in maternal brain (P<0.05). alpha Gi(1,2) and alpha Gi(3) subunits were not affected in mothers or fetuses as revealed by immunoblotting. mRNA levels coding these subunits were also unaffected in mothers and fetuses. On the other hand, forskolin- and forskolin plus guanosine-5'-O-(3-thiotriphosphate) (GTP gamma S)-stimulated adenylyl cyclase activity was decreased in maternal (P<.01) and fetal brain (P<.001). Furthermore, adenylyl cyclase inhibition elicited by N(6)-cyclohexyladenosine (CHA), a selective A(1)R agonist, was significantly decreased in both maternal (P<0.05) and fetal brain (P<.01), suggesting a desensitization of the A(1)R/adenylyl cyclase pathway. Therefore, these results suggest that R-PIA intake during pregnancy causes desensitization of the A(1)R-mediated inhibitory transduction pathway in both maternal and fetal brain, probably due to the decreased density of A(1)R at the cell surface. PMID- 19559761 TI - Interaction of elongation factor 1-alpha with leucine-rich repeat kinase 2 impairs kinase activity and microtubule bundling in vitro. AB - Autosomal dominant mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common genetic cause of late-onset Parkinson's disease. However, the regulators/effectors contributing to the (patho-)physiological functions of LRRK2 remain poorly defined. Here we show that human LRRK2 co-purifies/co immunoprecipitates with elongation factor 1-alpha (EF1A). Co-incubation of recombinant LRRK2 and EF1A significantly reduces the kinase activity of LRRK2, whereas its GTPase activity remains unchanged. In addition to its canonical role in mRNA translation, EF1A maintains stability of the microtubule cytoskeleton. In the present study, EF1A promotes microtubule assembly in an in vitro tubulin polymerization assay which is impaired by co-incubation with LRRK2 at sub stoichiometric concentrations. These findings suggest that the interaction between LRRK2 and EF1A may reciprocally modulate their physiological function. PMID- 19559762 TI - Gender and the use of hormonal contraception in women are not associated with cerebral cortical 5-HT 2A receptor binding. AB - Gender influences brain function including serotonergic neurotransmission, which may play a role in the well-known gender variations in vulnerability to mood and anxiety disorders. Even though hormonal replacement therapy in menopause is associated with globally increased cerebral 5-HT(2A) receptor binding it is not clear if gender or use of hormonal contraception exhibits associations with 5 HT(2A) receptor binding. We found no significant effect of gender on cortical 5 HT(2A) receptor binding (P=0.15, n=132). When adjusting for the personality trait neuroticism, known to be positively correlated to frontolimbic 5-HT(2A) receptor binding and to be more pronounced in women, again, the effect of gender was not significant (P=0.42, n=127). Also, the use of hormonal contraception (n=14) within the group of pre-menopausal women (total n=29) was not associated with cortical 5-HT(2A) receptor binding (P=0.31). In conclusion, neither gender, nor the use of hormonal contraception in premenopausal women was associated with cortical 5-HT(2A) receptor binding. PMID- 19559763 TI - Alterations in AMPA receptor phosphorylation in the rat striatum following acute and repeated cocaine administration. AB - Protein phosphorylation is an important mechanism for the posttranslational modulation of ionotropic glutamate receptors and is subject to regulation by changing synaptic inputs. In this study, we investigated the regulation of alpha amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor GluR1 subunit phosphorylation by cocaine exposure in the rat dorsal striatum in vivo. We found that acute cocaine challenge followed by 6 days of repeated systemic injections of cocaine (20 mg/kg once daily) enhanced the sensitivity of the GluR1 subunit in its phosphorylation at serine 831 (Ser831) in the dorsal striatum. This enhancement of the sensitivity of Ser831 phosphorylation was reduced, at the receptor and ion channel level, by blocking (1) group I metabotropic glutamate receptors (mGluRs), (2) N-methyl-D-aspartate receptors, and (3) L-type voltage operated Ca(2+) channels. Similar reduction of the enhancement was also induced, at the protein kinase level, by inhibiting (1) protein kinase C, (2) calcium/calmodulin-dependent protein kinases, and (3) c-Jun N-terminal kinases. In addition, inhibition of protein phosphatase 1/2A or calcineurin increased GluR1-Ser831 phosphorylation in the dorsal striatum of normal rats, whereas inhibition of these phosphatases did not further enhance the Ser831 phosphorylation in rats pretreated with 7 daily injections of cocaine. These data suggest that the phosphorylation of AMPA receptor GluR1 subunits at Ser831 is subject to upregulation by acute and repeated cocaine administration. Complex signaling integrations among glutamate receptors, Ca(2+) channels, protein kinases, and protein phosphatases participate in this upregulation. PMID- 19559764 TI - Dopamine D1 receptor-dependent modifications in the dopamine and cAMP-regulated phosphoprotein of Mr 32 kDa phosphorylation pattern in striatal areas of morphine sensitized rats. AB - Morphine sensitization is a model of latent, functionally inducible increase in dopamine D(1) receptor-mediated transmission, which may be unmasked by an external stimulus. Morphine-sensitized rats present dopamine D(1) receptor dependent stereotypies upon morphine challenge and resilience to unavoidable stress-induced behavioral deficits. This tonic increase in dopamine D(1) dopaminergic transmission is counter-adaptive to an enhanced mu-opioid receptor dependent signaling in striatal areas. Control and sensitized rats show a similar dopamine and cAMP-regulated phosphoprotein of M(r) 32 kDa (DARPP-32) phosphorylation pattern in striatal areas. Acute morphine administration induced an early increase and delayed decrease in phospho-threonine (Thr)34 DARPP-32 levels accompanied by a delayed increase in phospho-Thr75 DARPP-32 levels in the nucleus accumbens and caudate-putamen of sensitized rats, while it had no effects in control animals. The administration of a selective dopamine D(1) receptor antagonist (SCH 23390) before morphine challenge prevented the behavioral and neurochemical modifications in sensitized rats. 6-Methyl-2-(phenylethynyl) pyridine, a selective metabotropic glutamate receptor 5 (mGluR(5)) antagonist, administered 1 h after morphine challenge, prevented the delayed phosphorylation changes, but it had no effect when administered before challenge. Moreover, the DARPP-32 phosphorylation pattern in the caudate-putamen of sensitized rats after unavoidable stress exposure was studied. The stress-induced neurochemical modifications and their sensitivity to receptor antagonists were similar to those observed after acute morphine administration. In conclusion, these results suggest that in the experimental conditions used an increase in dopamine output in striatal areas is followed by a complex neurochemical pattern, in which the initial stimulation of dopamine D(1) receptors triggers a sequence of signaling events that lead to an mGluR(5)-mediated increase in phospho-Thr75 DARPP-32 levels. Since DARPP-32 phosphorylated in Thr75 inhibits cAMP-dependent protein kinase (PKA) activity, the final result is a decrease in the dopamine D(1) receptor-dependent phosphorylation events. PMID- 19559765 TI - Use of the Cognitive Performance Scale (CPS) to detect cognitive impairment in the acute care setting: concurrent and predictive validity. AB - The Cognitive Performance Scale (CPS) was initially designed to assess cognition in long term care residents. Subsequently, the CPS has also been used among in home, post-acute, and acute care populations even though CPS' clinimetric performance has not been studied in these settings. This study aimed to determine CPS agreement with the Mini Mental Status Exam (MMSE) and its predictive validity for institutionalization and death in a cohort (N=401) of elderly medical inpatients aged 75 years and over. Medical, physical and mental status were assessed upon admission. The same day, the patient's nurse completed the CPS by interview. Follow-up data were gathered from the central billing system (nursing home stay) and proxies (death). Cognitive impairment was present in 92 (23%) patients according to CPS (score >or= 2). Agreement with MMSE was moderate (kappa 0.52, P<.001). Analysis of discordant results suggested that cognitive impairment was overestimated by the CPS in dependent patients with comorbidities and depressive symptoms, and underestimated in older ones. During follow-up, subjects with abnormal CPS had increased risks of death (adjusted hazard ratio (adjHR) 1.7, 95% CI 1.0-2.8, P=.035) and institutionalization (adjHR 2.7, 95% CI 1.3-5.3, P=.006), independent of demographic, health and functional status. Interestingly, subjects with abnormal CPS were at increased risk of death only if they also had abnormal MMSE. The CPS predicted death and institutionalization during follow-up, but correlated moderately well with the MMSE. Combining CPS and MMSE provided additional predictive information, suggesting that domains other than cognition are assessed by professionals when using the CPS in elderly medical inpatients. PMID- 19559766 TI - Agroclavine potentiates hippocampal EEG effects of weak combined magnetic field in rats. AB - To learn whether the low flux density combined magnetic field (CMF) can affect rats pretreated with a dopamine agonist, the frequency spectra of cortical and hippocampal EEG were studied after intraperitoneal injection of agroclavine (50 microg/kg, Day 1) either alone (N=5) or combined with 30-min exposure to CMF (B(AC)=50 microT, f(AC)=20.9Hz, B(DC)=27.3 microT) (N=5) an hour post-injection. The CMF caused an immediate and robust increase in the hippocampal beta(2) activity (22.5-30.5Hz) vs. its value in the pre-exposure period. This effect persisted during 30-min post-exposure period. After vehicle injection (0.1% ethanol) on Day 10, the beta(2) activity increased gradually during the exposure to CMF and was at the pre-exposure level during the post-exposure period. In two rats, pretreated with the vehicle on Day 1, CMF increased a hippocampal beta(2) level. The effect was about half the amplitude and shorter in duration than that after agroclavine, with no effects in the post-exposure period. However, during the 2nd exposure to CMF, started 30 min later, cortical and hippocampal beta activity was significantly greater than that during the 1st. On Day 10, the 1st exposure was completely ineffective, while during the 2nd, the hippocampal beta(2) activity was increased gradually. These results demonstrate that dopaminergic activation can potentiate CMF-evoked EEG effects in the hippocampal beta(2) frequency band. PMID- 19559767 TI - Differential damage in the frontal cortex with aging, sporadic and familial Alzheimer's disease. AB - In order to understand relationships between executive and structural deficits in the frontal cortex of patients within normal aging or Alzheimer's disease, we studied frontal pathological changes in young and old controls compared to cases with sporadic (AD) or familial Alzheimer's disease (FAD). We performed a semi automatic computer assisted analysis of the distribution of beta-amyloid (Abeta) deposits revealed by Abeta immunostaining as well as of neurofibrillary tangles (NFT) revealed by Gallyas silver staining in Brodman areas 10 (frontal polar), 12 (ventro-infero-median) and 24 (anterior cingular), using tissue samples from 5 FAD, 6 sporadic AD and 10 control brains. We also performed densitometric measurements of glial fibrillary acidic protein, principal compound of intermediate filaments of astrocytes, and of phosphorylated neurofilament H and M epitopes in areas 10 and 24. All regions studied seem almost completely spared in normal old controls, with only the oldest ones exhibiting a weak percentage of beta-amyloid deposit and hardly any NFT. On the contrary, all AD and FAD cases were severely damaged as shown by statistically significant increased percentages of beta-amyloid deposit, as well as by a high number of NFT. FAD cases (all from the same family) had statistically more beta-amyloid and GFAP than sporadic AD cases in both areas 10 and 24 and statistically more NFT only in area 24. The correlation between the percentage of beta-amyloid and the number of NFT was significant only for area 24. Altogether, these data suggest that the frontal cortex can be spared by AD type lesions in normal aging, but is severely damaged in sporadic and still more in familial Alzheimer's disease. The frontal regions appear to be differentially vulnerable, with area 12 having the less amyloid burden, area 24 the less NFT and area 10 having both more amyloid and more NFT. This pattern of damage in frontal regions may represent a strong neuroanatomical support for the deterioration of attention and cognitive capacities as well as for the presence of emotional and behavioral troubles in AD patients. PMID- 19559768 TI - Metabolic clustering of a core collection of Indian ginseng Withania somnifera Dunal through DNA, isoenzyme, polypeptide and withanolide profile diversity. AB - Withania somnifera is one of the most important medicinal plants of Ayurveda and finds extensive uses in Indian traditional herbal preparations. In this investigation, selected accessions of the plant were examined for diversity through RAPDs, isoenzymes, polypeptide polymorphism and withanolide profiles. The accessions clustered together with respect to their characteristic profile of major withanolides and represented withaferin A, withanone, withanolide D or withanolide A rich groups. This level of phytochemical diversity as discrete chemotypes is widest and is being first ever documented to occur in Indian population of the plant. PMID- 19559769 TI - Simultaneous quantification of eight bioactive components of Houttuynia cordata and related Saururaceae medicinal plants by on-line high performance liquid chromatography-diode array detector-electrospray mass spectrometry. AB - An on-line high performance liquid chromatography (HPLC)-diode array detector (DAD)-electrospray ionization mass spectrometry (ESI-MS) method has been developed to quantify simultaneously eight bioactive chemical components in Houttuynia cordata Thunb and related Saururaceae medicinal plants. Simultaneous separation of these eight compounds was achieved on a C(18) analytical column with gradient elution of acetonitrile and 0.2% acetic acid (v/v) at a flow rate of 0.6 mL/min and being detected at 280 nm. These eight compounds were completely separated within 90 min. Good linear regression relationship (r(2)>0.9978) within test ranges was shown in all calibration curves. Good repeatabilty for the quantification of these eight compounds in H.cordata was also demonstrated in this method, with intra- and inter-day variations less than 3.0%. The method established was successfully applied to quantify eight bioactive compounds in closely related species of H.cordata, which provides a new basis for quality assessment of H.cordata. PMID- 19559770 TI - Effects of isopulegol on pentylenetetrazol-induced convulsions in mice: possible involvement of GABAergic system and antioxidant activity. AB - The present study investigated the effects of isopulegol, a monoterpene alcohol, in PTZ-induced convulsions and verified possible involved mechanisms. Saline, isopulegol or diazepam were intraperitonealy injected 30 min before PTZ. The latency for development of convulsions and mortality, as well as the mortality protection percentage was recorded. For investigating the involvement of GABAergic system, flumazenil was utilized. The activity of antioxidant enzyme catalase as well as the levels of reduced glutathione and lipid peroxidation were measured in brain hippocampus. Similarly to diazepam, isopulegol significantly prolonged the latency for convulsions and mortality of mice. All animals were protected against mortality at higher dose of isopulegol. Flumazenil pretreatment decreased the prolongation of seizure latency induced by both diazepam and isopulegol, although it was not able to reverse the latency and protection percent for mortality. Isopulegol also significantly prevented PTZ-induced increase in lipid peroxidation, preserved catalase activity in normal levels, and prevented the PTZ-induced loss of GSH in hippocampus of mice. These results suggest that the anticonvulsant and bioprotective effects of isopulegol against PTZ-induced convulsions are possibly related to positive modulation of benzodiazepine-sensitive GABA(A) receptors and to antioxidant properties. PMID- 19559771 TI - Acute restraint stress produces behavioral despair in weanling rats in the forced swim test. AB - Stressful experiences in the rat during early life increase the vulnerability to later signs of behavioral despair in adulthood, reflected in increased immobility in the forced swim test (FST). However, the possible immediate effects of stress in weanling rats have only been partially described. The present study tested whether a single session of mild restraint stress modifies immobility in the FST in 21-day-old Wistar rats. After evaluating any possible changes in locomotion using the open field test (OFT), the latency and total duration of immobility were assessed in a single FST session. Regardless of gender, mild restraint stress significantly reduced crossings in the OFT, shortened the latency to the first period of immobility, and increased immobility in the FST compared with a control group devoid of stress. We conclude that a single mild physical stress session, as early as postnatal day 21, produces signs of behavioral despair. PMID- 19559773 TI - Identification of GATA3 binding sites in Jurkat cells. AB - Determining binding sites of transcription factors is important for understanding the transcriptional control of target genes. Although a transcription factor GATA3 plays a pivotal role in Th2 lymphocyte development, its physiological role is not clearly defined because the target genes remain largely unknown. In this study, we modified chromatin immunoprecipitation (ChIP), and isolated 121 GATA3 binding sites and 83 different annotated target genes. Re-ChIP analysis using anti-GATA3 and anti-RNA polymerase II mAbs and chromosome conformation capture assay demonstrate that GATA3-bound fragments interact with basal transcriptional units of target genes. GATA3 regulation of target genes under the control of binding fragments was confirmed by reporter assay and quantification of target gene mRNA expression in the presence of GATA inhibitor or short interfering RNA against GATA3. These data demonstrate that GATA3 binds to regulatory elements and controls target gene expression through physical interaction with core promoter regions. PMID- 19559772 TI - Identification and characterisation of a novel antisense non-coding RNA from the RBM5 gene locus. AB - Previous work from our lab identified a 326 base-pair (bp) cDNA, termed Je2, which mapped to the antisense strand of intron 6 of the putative tumour suppressor gene RBM5/LUCA-15/H37, and functioned as an apoptosis suppressor. The purpose of the work described herein was to determine if Je2 is part of a larger transcript, to clone that transcript and to examine its ability to modulate RBM5 expression. Northern blot analyses in conjunction with strand-specific reverse transcription and PCR revealed two novel transcripts, one antisense and one sense, that included Je2 as well as RBM5 intron 4 sequence. Using rapid amplification of cDNA ends (RACE), a novel 1.4 kb product including Je2 and intron 4 was cloned. In vitro transcription/translation did not result in the production of any protein product, from either strand. Genomic DNA analysis revealed the presence of a putative promoter region 5' to Je2, suggesting that the cloned 1.4 kb RACE product represents an antisense transcript that initiates within intron 6 and terminates within intron 4 of the RBM5 gene. This novel antisense, non-coding RNA was termed LUST, for LUCA-15-specific transcript. Ectopic overexpression of LUST coincided with elevated expression of the full length RBM5+5+6 alternative RBM5 RNA splice variant, and reduced expression of the truncated, cytotoxic RBM5+5+6t/Clone 26 alternative RBM5 RNA splice variant. A model is proposed whereby LUST functions co-transcriptionally to mask a sense strand regulatory sequence, common to both RBM5+5+6 and RBM5+5+6t/Clone 26 transcripts, that when unmasked results in premature termination of RBM5+5+6, thereby generating the cytotoxic truncated product, RBM5+5+6t/Clone 26. These results suggest that LUST is a novel, functional, non-coding RNA that plays a role in determining the apoptotic fate of a cell by regulating the expression of RBM5 splice variants. PMID- 19559775 TI - Liposome coated with low molecular weight chitosan and its potential use in ocular drug delivery. AB - In this study liposome coated with low molecular weight chitosan (LCH) was proposed and investigated its in vitro and in vivo properties, and its potential use in ocular drug delivery was evaluated. LCH with a molecular weight of 8kDa was prepared and coated on liposome loaded with diclofenac sodium. The LCH coating changed the liposome surface charge and slightly increased its particle size, while the drug encapsulation was not affected. After coating, the liposome displayed a prolonged in vitro drug release profile. LCH coated liposome also demonstrated an improved physicochemical stability at 25 degrees C in a 30-day storage period. The ocular bioadhesion property was evaluated by rabbit in vivo precorneal retention, and LCH coated liposome achieved a significantly prolonged retention compared with non-coated liposome or drug solution. The LCH coating also displayed a potential penetration enhancing effect for transcorneal delivery of the drug. In the ocular tolerance study, no irritation or toxicity was caused by continual administration of LCH coated liposome in a total period of 7 days. In conclusion, the LCH coating significantly modified the properties of liposome and brought a series of notable advantages for ocular drug delivery. PMID- 19559774 TI - Alteration of transcriptional profile in human bronchial epithelial cells induced by cigarette smoke condensate. AB - Despite the significance of cigarette smoke for carcinogenesis, the molecular mechanisms that lead to increased susceptibility of human cancers are not well understood. In our present study, the oncogenic transforming effects of cigarette smoke condensate (CSC) were examined using papillomavirus-immortalized human bronchial epithelial cells (BEP2D). Growth kinetics, saturation density, resistance to serum-induced terminal differentiation, anchorage-independent growth and tumorigenicity in nude mice were used to investigate the various stages of transformation in BEP2D cells. Illumina microarray platforms were used to explore the CSC-induced alteration of global mRNA expression profiles of the earlier period and the advanced stage of CSC-treated BEP2D cells. We showed here that a series of sequential steps arose among CSC-treated immortalized human bronchial epithelial cells, including altered growth kinetics, resistance to serum-induced terminal differentiation, and anchorage-independence growth. In the earlier period of CSC treatment, 265 genes were down-regulated and 63 genes were up-regulated, respectively, and in the advanced stage of CSC treatment, 313 genes were down-regulated and 145 genes were up-regulated, respectively. Notably, among those genes, the expression of some of imprinted genes such as IGF2, NDN, H19 and MEG3 were all silenced or down-regulated in CSC-treated cells. These genes reactivated after 5 microM 5-aza-2-deoxycytidine (5-aza-dC) treatment. These results demonstrated that long-term treatment of human bronchial epithelial cells with CSC may adversely affect their genetic and epigenetic integrity and lead to further transformation. PMID- 19559776 TI - Pharmacopeial compliance of fish oil-containing parenteral lipid emulsion mixtures: Globule size distribution (GSD) and fatty acid analyses. AB - Recently, the United States Pharmacopeia (USP) has established Chapter 729 with GSD limits for all lipid emulsions where the mean droplet size (MDS) must be <500 nm and the percent of fat larger than 5 microm (PFAT(5)) must be <0.05%, irrespective of the final lipid concentration. As well, the European Pharmacopeia (EP) Monograph no. 1352 specifies n3-fatty acid (FA) limits (EPA+DHA> or =45%; total n3 or T-n3> or =60%) for fish oil. We assessed compliance with USP physical and EP chemical limits of two fish oil-containing lipid emulsion mixtures. All lipid emulsions passed USP 729 limits. No samples tested had an MDS >302 nm or a PFAT(5) value >0.011%. Only one product met EP limits while the other failed. All emulsions tested were extremely fine dispersions and easily met USP 729 GSD limits. The n3-FAs profiles were lower in one, despite being labeled to contain 50% more fish oil than the other product. This latter finding suggests the n3-FA content of the fish oil source and/or the applied manufacturing processes in these products is different. PMID- 19559777 TI - Antitumor and immunomodulatory activity of polysaccharides from the roots of Actinidia eriantha. AB - AIM OF THE STUDY: The roots of Actinidia eriantha Benth (Actinidiaceae) have been used for cancers in the Chinese folk medicine. The present study aimed at evaluating the antitumor potentials of the polysaccharides from the roots of Actinidia eriantha and elucidating their immunological mechanisms by determining the effects on the growth of tumor transplanted in mice and the immune response in tumor-bearing mice. MATERIALS AND METHODS: The total polysaccharide AEP and fours purified polysaccharides AEPA, AEPB, AEPC and AEPD were isolated and purified from the roots of Actinidia eriantha by hot water extraction, ethanol precipitation, dialysis and gel filtration. Their effects on the growth of mouse transplantable tumor, splenocyte proliferation, the activity of natural killer (NK) cells and cytotoxic T lymphocytes (CTL), production of cytokines from splenocytes, and serum antigen-specific antibody levels in tumor-bearing mice were measured. RESULTS: AEP and four purified polysaccharides could not only significantly inhibit the growth of mouse transplantable tumor, but also remarkably promote splenocytes proliferation, NK cell and CTL activity, IL-2 and IFN-gamma production from splenocytes, and serum antigen-specific antibody levels in tumor-bearing mice. CONCLUSIONS: The antitumor activity of AEP and four purified polysaccharides might be achieved by improving immune response, and the composition of the monosaccharides, uronic acid contents and molecular weight could affect their antitumor and immunomodulatory activity. PMID- 19559778 TI - Glycyrretinic acid blocks cardiac sodium channels expressed in Xenopus oocytes. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Licorice has been used to treat many ailments including cardiovascular disorders in China for long time. Recent studies have shown that the cardiac actions of licorice have been attributed to its active component, glycyrretinic acid (GA). However, its mechanism remains poorly understood. AIM OF THE STUDY: The effects of GA on the cardiac sodium currents (I(Na)), L-type calcium currents (I(Ca,L)) and hyperpolarization-activated inward currents (I(f)) were investigated. MATERIALS AND METHODS: Human isoforms of wild type and DeltaKPQ-mutant type sodium channels were expressed in Xenopus oocytes, and the resulting currents (peak and late I(Na)) were recorded using a two microelectrode voltage-clamp technique. A perforated patch clamp technique was employed to record I(Ca,L) and I(f) from isolated rabbit sinoatrial node pacemaker cells. RESULTS: GA inhibited peak I(Na) (33% at 90 microM) and late I(Na) (72% at 90 microM), but caused no significant effects on I(Ca,L) and I(f). CONCLUSION: GA blocked cardiac sodium currents, particularly late I(Na.) Our findings might help to understand the traditional use of licorice in the treatment of cardiovascular disorders, because reduction of sodium currents (particularly late I(Na)) would be expected to provide protection from Na(+) induced Ca(2+) overload and cell damage. PMID- 19559779 TI - Anti-inflammatory effects of the extract of indigo naturalis in human neutrophils. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Indigo naturalis is used by traditional Chinese medicine to treat various inflammatory diseases. AIM OF THE STUDY: Topical indigo naturalis ointment showed efficacy in treating psoriasis in our previous clinical studies. In this study, we investigated the anti-inflammatory effects of the extract of indigo naturalis (QD) and its main components indirubin, indigo, and tryptanthrin in human neutrophils. MATERIALS AND METHODS: Superoxide anion (O2(. )) generation and elastase release were measured by spectrophotometry. Some important signals including mitogen-activated protein kinase (MAPK), cAMP, and calcium were studied by Western blot analysis, an enzyme immunoassay, and spectrofluorometry. RESULTS: QD significantly inhibited O2(.-) generation and elastase release in formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP)-activated human neutrophils in a concentration-dependent fashion, while neither indirubin, indigo, nor tryptanthrin produced a comparable result. QD attenuated the FMLP induced phosphorylation of extracellular regulated kinase, p38 MAPK, and c-Jun N terminal kinase. Furthermore, QD inhibited calcium mobilization caused by FMLP. However, QD did not affect cellular cAMP levels. On the other hand, neither indirubin, indigo, nor tryptanthrin produced similar changes in human neutrophils. CONCLUSIONS: Taken collectively, these findings indicate that QD, but not indirubin, indigo, or tryptanthrin, inhibited O2(.-) generation and elastase release in FMLP-induced human neutrophils, which was at least partially mediated by the inhibition of MAPK activation and regulation of calcium mobilization. PMID- 19559780 TI - Adenosine deaminase activity, lipid peroxidation and astrocyte responses in the cerebral cortex of rats after neonatal hypoxia ischemia. AB - Hypoxia ischemia (HI) is a common cause of damage in the fetal and neonatal brain. Lifelong disabilities such as cerebral palsy, epilepsy, behavioral and learning disorders are some of the consequences of brain injury acquired in the perinatal periods. Inflammation and formation of free radicals appear to play key roles in neonatal HI. The aim of this study was to describe the chronological sequence of adenosine deaminase (ADA) activity, the oxidative damage changes and astrocyte response using the classic model of neonatal HI. We observed an increase in the activity of ADA and lipid peroxidation in the cerebral cortex 8 days after neonatal HI. This was accompanied by a GFAP-positive, and the degree of brain damage was determined histochemically by hematoxylin-eosin (HE). Taking into account the important anti-inflammatory role of adenosine, ADA may provide an efficient means for scavenging cell-surrounding adenosine and play an important part in subsequent events of neonatal HI in association with GFAP reactive gliosis. The present investigation showed that neonatal HI causes the increase of free radicals and significant damage in the cerebral cortex. The increase in ADA activity may reflect the activation of the immune system caused by HI because the morphological analysis exhibited a lymphocytic infiltration. PMID- 19559781 TI - Prenatal auditory stimulation alters the levels of CREB mRNA, p-CREB and BDNF expression in chick hippocampus. AB - Prenatal auditory stimulation influences the development of the chick auditory pathway and the hippocampus showing an increase in various morphological parameters as well as expression of calcium-binding proteins. Calcium regulates the activity of cyclic adenosine monophosphate-response element binding (CREB) protein. CREB is known to play a role in development, undergo phosphorylation with neural activity as well as regulate transcription of BDNF. BDNF is important for the survival of neurons and regulates synaptic strength. Hence in the present study, we have evaluated the levels of CREB mRNA and protein along with p-CREB protein as well as BDNF mRNA and protein levels in the chick hippocampus at embryonic days (E) 12, E16, E20 and post-hatch day (PH) 1 following activation by prenatal auditory stimulation. Fertilized eggs were exposed to species-specific sound or sitar music (frequency range: 100-6300Hz) at 65dB levels for 15min/h over 24h from E10 till hatching. The control chick hippocampus showed higher CREB mRNA and p-CREB protein in the early embryonic stages, which later decline whereas BDNF mRNA and BDNF protein levels increase until PH1. The CREB mRNA and p CREB protein were significantly increased at E12, E16 and PH1 in the auditory stimulated groups as compared to control group. A significant increase in the level of BDNF mRNA was observed from E12 and the protein expression from E16 onwards in both auditory stimulated groups. Therefore, enhanced phosphorylation of CREB during development following prenatal sound stimulation may be responsible for cell survival. Increased levels of p-CREB again at PH1 may trigger synthesis of proteins necessary for synaptic plasticity. Further, the increased levels of BDNF may also help in regulating synaptic plasticity. PMID- 19559782 TI - Expression levels of NF-Y target genes changed by CDKN1B correlate with clinical prognosis in multiple cancers. AB - CDK inhibitors CDKN1B (p27) and CDKN2A (p16) inhibit cell cycle progression. A lower expression level of only p27 has been correlated with poorer prognosis in various types of clinical cancers. The difference may be the result of distinct genes downstream of these CDK inhibitors. Here, we report that NF-Y transcription factor-targeted genes specifically down-regulated by p27 correlate with poor prognosis in multiple tumor types. We performed mRNA expression profiling in HCT116 cells over-expressing either p16 or p27 and identified their regulatory genes. In silico transcription factor prediction indicated that most of the genes specifically down-regulated by p27 are controlled by NF-Y. Under the hypothesis that NF-Y-targeted genes are responsible for poor prognosis, we predicted prognosis in four types of cancer based on genes with the NF-Y motif, and found a significant association between the expression of NF-Y-targeted genes and poor prognosis. PMID- 19559783 TI - A genome-wide survey of copy number variations in Han Chinese residing in Taiwan. AB - Copy number variation (CNV) is a form of DNA sequence variation in the human genome. CNVs can affect expression of nearby and distant genes, and some of them might cause certain phenotypic differences. CNVs vary slightly in location and frequency among different populations. Because currently-available CNV information from Asian population was limited to fewer small-scale studies with only dozens of subjects, a high-resolution CNV survey was conducted using a large number of Han Chinese in this study. The Illumina HumanMap550K single-nucleotide polymorphism array was used to identify CNVs from 813 unrelated Han Chinese residing in Taiwan. A total of 365 CNV regions were identified in this population, and the average size of the CNV regions was 235 kb (covering a total of 2.86% of the human genome), and 67 (18.4%) were newly-discovered CNV regions. Two hundred and seventy-nine CNV regions (76%) were verified from 304 randomly selected samples by Affymetrix 500K GeneChip and qPCR experiments. These regions contain 1029 genes, some of which are associated with diseases. Consistent with previous studies, most CNVs were rare structural variations in the human genome, and only 64 regions (17.5%) had a CNV allele frequency greater than 1%. Our discovery of 67 new CNV regions indicates that previous CNV coverage of the human genome is incomplete and there is diversity among different ethnic populations. The comprehensive knowledge of CNVs in the human genome is very important and useful in further genetic studies. PMID- 19559784 TI - Activation of microglia by neuronal activity: results from a new in vitro paradigm based on neuronal-silicon interfacing technology. AB - Cognition and behavior primarily arise from the communication that occurs between brain cells. By using photoconductive stimulation to trigger localized regions of neuronal action potentials and astrocyte Ca(2+) waves in dissociated rat hippocampal cultures, we can directly study microglia behavior in response to physiological and pathological levels of activity. Connections between neurons can be modified by microglia, which regulate gap junctions and synapses through secretion of proteins such as cytokines, proteases and neurotrophic factors. Activated microglia participate in bidirectional communication with the excitable tissues that they support. Through feedback from the many ion channels and surface receptors they express, microglia are informed of neuronal and astrocytic activity that may indicate disruption in the homeostasis of the CNS. Such disturbances alert microglia to locations of such activity and promote their transformation into a reactive state, in which they perform adaptive functions that can be either neuroprotective, neurotoxic, or neuromodulatory. Under physiological conditions, normal brain activity has the effect of suppressing microglia inflammatory responses. This report summarizes available data about the interaction of microglia and brain activity and presents a new in vitro paradigm to study the mechanisms involved. We propose that photoconductive stimulation is a powerful tool for studying the cellular and molecular mechanisms underlying the dynamic interactions between neurons, astrocytes and microglia. PMID- 19559786 TI - Rapid changes on nitrinergic system in female mouse hippocampus during the ovarian cycle. AB - Fluctuating levels of estradiol and progesterone during the estrous cycle may induce structural changes in several brain nuclei including the hippocampus, where some neurons express estrogen receptors. Nitric oxide plays a wide range of functions in the nervous system generally by acting as a neurotransmitter-like molecule. It has been demonstrated that long-term treatments with estradiol in ovariectomized females and with testosterone in castrated males induce neuronal nitric oxide synthase (nNOS) expression in rat hypothalamus, whereas changes in nNOS immunoreactivity or in associated NADPH-diaphorase activity were observed both in hypothalamus and in amygdala during different phases of estrous cycle. Estradiol could induce nNOS expression in several brain regions in rodents. Therefore, to clarify if the hippocampal NO producing system is a target for gonadal hormones in physiological conditions, we have investigated the effects of estrous cycle in the expression of nNOS immunoreactivity on two-month-old intact female mice. Immunoreactive cells were observed in all hippocampal subregions: the higher number was detected in the pyramidal layer of CA1 region and in polymorph layer of dentate gyrus. The number of nNOS positive neurons fluctuates during the estrous cycle, reaching its peak during proestrus and metaestrus, and these variations were statistically significant in CA1, CA2 and CA3 regions. These results suggest that the nitrinergic system is a target for estrogen action in the hippocampus, and that this action may take place in physiological conditions according to the short-term variations of gonadal hormones during the estrous cycle. PMID- 19559787 TI - Special relationship between sterols and oxygen: were sterols an adaptation to aerobic life? AB - A fascinating link between sterols and molecular oxygen (O(2)) has been a common thread running through the fundamental work of Konrad Bloch, who elucidated the biosynthetic pathway for cholesterol, to recent work supporting a role of sterols in the sensing of O(2). Synthesis of sterols by eukaryotes is an O(2)-intensive process. In this review, we argue that increased levels of O(2) in the atmosphere not only made the evolution of sterols possible, but that these sterols may in turn have provided the eukaryote with an early defence mechanism against O(2). The idea that nature crafted sterols as a feedback loop to adapt to, or help protect against, the hazards of O(2) is novel and enticing. We marshal several lines of evidence to support this thesis: (1) coincidence of atmospheric O(2) and sterol evolution; (2) sterols regulate O(2) entry into eukaryotic cells and organelles; (3) sterols act as O(2) sensors across eukaryotic life; (4) sterols serve as a primitive cellular defence against O(2) (including reactive oxygen species). Therefore, sterols may have evolved in eukaryotes partially as an adaptive response to the rise of terrestrial O(2), rather than merely as a consequence of it. PMID- 19559785 TI - Positive affect and inflammation during radiation treatment for breast and prostate cancer. AB - There is growing evidence that positive affect may influence health and immune function, although few studies have examined links between positive affect and immune processes in clinical populations. The purpose of this study was to examine whether positive affect is associated with changes in proinflammatory cytokines in cancer patients undergoing radiation treatment. Subjects were 50 individuals with early-stage breast and prostate cancer who completed psychosocial questionnaires and provided blood samples at seven time points before, during, and after radiation treatment. Positive affect was assessed before treatment onset using the CES-D (Center for Epidemiological Studies Depression Scale). Blood samples were assayed for serum levels of proinflammatory cytokines IL-1beta and IL-6. Patients with higher levels of positive affect before treatment exhibited higher mean levels of IL-1beta and IL-6 during radiation treatment (all ps<.05). Results suggest that positive affect enhances the acute inflammatory response to radiation treatment, perhaps facilitating tissue repair processes. PMID- 19559788 TI - Superoxide radical protects liposome-contained cytochrome c against oxidative damage promoted by peroxynitrite and free radicals. AB - The effects of nitrosative species on cyt c structure and peroxidase activity were investigated here in the presence of O(2)(*-) and anionic and zwitterionic vesicles. Nitrosative species were generated by 3-morpholinesydnonymine (SIN1) decomposition, using cyt c heme iron and/or molecular oxygen as electron acceptor. Far- and near-UV CD spectra of SIN1-treated cyt c revealed respectively a slight decrease of alpha-helix content (from 39 to 34%) and changes in the tryptophan structure accompanied by increased fluorescence. The Soret CD spectra displayed a significant decrease of the positive signal at 403 nm. EPR spectra revealed the presence of a low-spin cyt c form (S=1/2) with g(1)=2.736, g(2)=2.465, and g(3)=2.058 after incubation with SIN1. These data suggest that the concomitant presence of NO(*) and O(2)(*-) generated from dissolved oxygen, in a system containing cyt c and liposomes, promotes chemical and conformational modifications in cyt c, resulting in a hypothetical bis-histidine hexacoordinated heme iron. We also show that, paradoxically, O(2)(*-) prevents not only membrane lipoperoxidation by peroxide-derived radicals but also oxidation of cyt c itself due to the ability of O(2)(*-) to reduce heme iron. Finally, lipoperoxidation measurements showed that, although it is a more efficient peroxidase, SIN1 treated cyt c is not more effective than native cyt c in promoting damage to anionic liposomes in the presence of tert-ButylOOH, probably due to loss of affinity with negatively charged lipids. PMID- 19559789 TI - From formamide to RNA: the roles of formamide and water in the evolution of chemical information. AB - In pursuing the origin of informational polymers, we followed the assumption that their spontaneous formation could only have occurred: (i) if all the components were present at the same site and in the same reaction, and (ii) if the thermodynamics of the processes involved favored a polymerized over a monomeric state of the precursors. A plausible scenario satisfying both assumptions is provided. PMID- 19559790 TI - Caspase-3 is activated and rapidly released from human umbilical vein endothelial cells in response to lipopolysaccharide. AB - Endothelial cell injury/dysfunction is considered to play a critical role in the pathogenesis of severe sepsis and septic shock. Although it is considered that endothelial cell apoptosis is involved in endothelial injury/dysfunction, physiological involvement remains ambiguous since the induction of apoptosis requires the inhibition of endogenous apoptosis inhibitors. Here we show that caspase-3 activation, a biological indicator of apoptosis, is observed in response to lipopolysaccharide (LPS) stimulation even under the influence of endogenous apoptosis inhibitors, and that activated caspase-3 is rapidly released from human umbilical vein endothelial cells (HUVEC). In the presence of cycloheximide (CHX), an increase in intracellular caspase-3/7 activity in response to LPS was not detected in HUVEC up to 24 h following stimulation even in the presence of LPS-binding protein (LBP), soluble CD14 and soluble MD-2, whereas the decrease in cell viability and increase in release of the cellular enzyme lactate dehydrogenase (LDH) were observed in a soluble CD14/LBP-dependent manner. On the other hand, even in the absence of CHX, a significant increase in caspase-3/7 activity and a cleaved caspase-3 fragment with a slight increase in LDH release was observed in culture supernatants in response to LPS. This increase in caspase-3/7 activity was observed even when LDH release was undetected. These results indicate that caspase-3 is activated by LPS under physiological conditions and suggest that HUVEC escape from cell death by rapidly releasing activated caspase-3 into extracellular space. Failure of this escape mechanism may result in endothelial injury/dysfunction. PMID- 19559791 TI - Selective removal of stratum corneum by microdermabrasion to increase skin permeability. AB - This study sought to determine if microdermabrasion can selectively remove stratum corneum to increase skin permeability. Although, microdermabrasion has been used for cosmetic treatment of skin for decades, no study has assessed the detailed effects of microdermabrasion conditions on the degree of skin tissue removal. Therefore, we histologically characterized the skin of rhesus macaques and human volunteers after microdermabrasion at different conditions. Using mobile tip microdermabrasion, an increase in the number of treatment passes led to greater tissue removal ranging from minimal effects to extensive damage to deeper layers of the skin. Of note, these data showed for the first time that at moderate microdermabrasion conditions selective yet full-thickness removal of stratum corneum could be achieved with little damage to deeper skin tissues. In the stationary mode of microdermabrasion, selective stratum corneum removal was not observed, but micro-blisters could be seen. Similar tissue removal trends were observed in human volunteers. As proof of concept for drug delivery applications, a model fluorescent drug (fluorescein) was delivered through microdermabraded skin and antibodies were generated against vaccinia virus after its topical application in monkeys. In conclusion, microdermabrasion can selectively remove full-thickness stratum corneum with little damage to deeper tissues and thereby increase skin permeability. PMID- 19559792 TI - The effect of rosiglitazone on asymmetric dimethylarginine (ADMA) in critically ill patients. AB - Asymmetric dimethylarginine (ADMA) plays a crucial role in the arginine-nitric oxide pathway. Critically ill patients have elevated levels of ADMA which proved to be a strong and independent risk factor for ICU mortality. The aim of this study was to investigate the effect of the peroxisome proliferator-activated receptor (PPAR)-gamma agonist rosiglitazone on ADMA plasma levels in critically ill patients. In a randomized controlled pilot study, ADMA, arginine and symmetric dimethylarginine (SDMA) were measured in 21 critically ill patients on the intensive care unit (ICU). Twelve patients received 4mg rosiglitazone once a day for a maximum of 6 weeks or until discharge or death. Nine patients served as control patients. In addition, total sequential organ failure assessment (SOFA score), kidney function and liver function were determined. Compared to the ADMA levels of healthy individuals as specified in earlier studies, ADMA plasma levels of critically ill patients were significantly higher (0.42+/-0.06 versus 0.73+/ 0.2micromol/L, respectively; p<0.001). Both ADMA (B=3.5; 95% CI: 0.5-6.5; p=0.023) and SDMA (B=1.7; 95% CI: 0.7-2.7; p=0.001) were independently related to SOFA scores. Overall, rosiglitazone treatment had no effect on ADMA levels, which only significantly differed between the rosiglitazone and control groups at day 7 (p=0.028). The SOFA score in the rosiglitazone group was lower compared to the control group but the difference was only statistically significant at day 10 (p=0.01). In conclusion, in critically ill patients plasma ADMA levels were elevated and associated with the extent of multiple organ failure, but no significant ADMA-lowering effect of the PPAR-gamma agonist rosiglitazone was observed. PMID- 19559793 TI - Correlates of arterial stiffness in an ageing population: role of asymmetric dimethylarginine. AB - A number of previous investigators have demonstrated that arterial augmentation index (AIx), a measure of apparent arterial stiffness, reflects in part vascular endothelial function, and that AIx is modulated by nitric oxide (NO) responses. We evaluated AIx in a population of 253 ageing subjects (mean age 63.4+/-6 (standard deviation, SD) years) and its relationship to (i) plasma levels of asymmetric dimethylarginine (ADMA), a marker and mediator of vascular endothelial dysfunction and (ii) the ratio of ADMA to its non-metabolised enantiomer symmetric dimethylarginine (SDMA), an inverse index of ADMA metabolic clearance. Evaluation was performed by univariate followed by multivariate analyses. On multivariate analyses, both ADMA (beta=0.16, p=0.01) and ADMA:SDMA (beta=0.21, p<0.001) ratio were significant direct correlates of AIx. Other significant correlates of AIx on multivariate analysis were: use of angiotensin-converting enzyme inhibitors/angiotensin-receptor blockers (ACEi/ARB) (beta=-0.24, p=0.004), smoking history (beta=0.15, p=0.007), male gender (beta=-0.38, p<0.001), creatinine clearance (CrCL) (beta=-0.25, p<0.001), and history of hypertension (beta=0.17, p=0.04). We conclude that (1) endothelial dysfunction engendered by impairment of NO synthesis may represent the basis for increased arterial stiffness in ageing individuals and (2) the fundamental biochemical anomaly may be impairment of ADMA clearance. These pathophysiological factors are likely to be relevant to optimize therapy to ameliorate disorders of arterial compliance in the ageing population. PMID- 19559794 TI - 3D structure of phosphofructokinase from Pichia pastoris: Localization of the novel gamma-subunits. AB - The largest and one of the most complex ATP-dependent allosteric phosphofructokinase (Pfk) has been found in the methylotrophic yeast, Pichia pastoris. The enzyme is a hetero-oligomer ( approximately 1MDa) composed of three distinct subunits (alpha, beta and gamma) with molecular masses of 109, 104 and 41kDa, respectively. While the alpha- and beta-subunits show sequence similarities to other phosphofructokinase subunits, the gamma-subunit does not show high homology to any known protein in the databases. We have determined the first quaternary structure of P. pastoris phosphofructokinase by 3D electron microscopy. Random conical techniques and tomography have been instrumental to ascertain the quality of the sample preparations for structural studies and to obtain a reliable 3D structure. The final reconstruction of P. pastoris Pfk resembles its yeast counterparts with four additional densities, assigned to four gamma-subunits, bridging the N-terminal domains of the four pairs of alpha- and beta-subunits. Our data has evidenced novel interactions between the gamma- and the alpha-subunits comparable in intensity to the interactions, shown by cross linking and limited proteolytic degradation experiments, between the gamma- and beta-subunits. The structural data provides clear insights into the allosteric fine-tuned regulation of the enzyme by ATP and AMP observed in this yeast species. PMID- 19559795 TI - Change-driven cortical activation in multisensory environments: an MEG study. AB - The quick detection of dynamic changes in multisensory environments is essential to survive dangerous events and orient attention to informative events. Previous studies have identified multimodal cortical areas activated by changes of visual, auditory, and tactile stimuli. In the present study, we used magnetoencephalography (MEG) to examine time-varying cortical processes responsive to unexpected unimodal changes during continuous multisensory stimulation. The results showed that there were change-driven cortical responses in multimodal areas, such as the temporo-parietal junction and middle and inferior frontal gyri, regardless of the sensory modalities where the change occurred. These multimodal activations accompanied unimodal activations, both of which in general had some peaks within 300 ms after the changes. Thus, neural processes responsive to unimodal changes in the multisensory environment are distributed at different timing in these cortical areas. PMID- 19559796 TI - Improvement of brain segmentation accuracy by optimizing non-uniformity correction using N3. AB - Smoothly varying and multiplicative intensity variations within MR images that are artifactual, can reduce the accuracy of automated brain segmentation. Fortunately, these can be corrected. Among existing correction approaches, the nonparametric non-uniformity intensity normalization method N3 (Sled, J.G., Zijdenbos, A.P., Evans, A.C., 1998. Nonparametric method for automatic correction of intensity nonuniformity in MRI data. IEEE Trans. Med. Imag. 17, 87-97.) is one of the most frequently used. However, at least one recent study (Boyes, R.G., Gunter, J.L., Frost, C., Janke, A.L., Yeatman, T., Hill, D.L.G., Bernstein, M.A., Thompson, P.M., Weiner, M.W., Schuff, N., Alexander, G.E., Killiany, R.J., DeCarli, C., Jack, C.R., Fox, N.C., 2008. Intensity non-uniformity correction using N3 on 3-T scanners with multichannel phased array coils. NeuroImage 39, 1752-1762.) suggests that its performance on 3 T scanners with multichannel phased-array receiver coils can be improved by optimizing a parameter that controls the smoothness of the estimated bias field. The present study not only confirms this finding, but additionally demonstrates the benefit of reducing the relevant parameter values to 30-50 mm (default value is 200 mm), on white matter surface estimation as well as the measurement of cortical and subcortical structures using FreeSurfer (Martinos Imaging Centre, Boston, MA). This finding can help enhance precision in studies where estimation of cerebral cortex thickness is critical for making inferences. PMID- 19559797 TI - Spatial smoothing hurts localization but not information: pitfalls for brain mappers. AB - Op de Beeck (Op de Beeck, H., 2009. Against hyperacuity in brain reading: Spatial smoothing does not hurt multivariate fMRI analyses? Neuroimage) challenges the possibility of extracting information from subvoxel representations via random biases associated with voxel sampling, the hypothesis proposed by Kamitani and Tong (Kamitani, Y., Tong, F., 2005. Decoding the visual and subjective contents of the human brain. Nat. Neurosci. 8, 679-685). Here, we show that his results provide no evidence against the possibility, being consistent with both of the subvoxel and supravoxel representation models. Classification of spatially smoothed fMRI data is not an effective means to probe into information sources for multivoxel decoding, since smoothing does not hurt the information contents of multivoxel patterns. We point out the danger of interpreting multivoxel decoding results based on intuitions guided by the conventional brain mapping paradigm. PMID- 19559798 TI - Bayesian multi-modal model comparison: a case study on the generators of the spike and the wave in generalized spike-wave complexes. AB - We present a novel approach to assess the networks involved in the generation of spontaneous pathological brain activity based on multi-modal imaging data. We propose to use probabilistic fMRI-constrained EEG source reconstruction as a complement to EEG-correlated fMRI analysis to disambiguate between networks that co-occur at the fMRI time resolution. The method is based on Bayesian model comparison, where the different models correspond to different combinations of fMRI-activated (or deactivated) cortical clusters. By computing the model evidence (or marginal likelihood) of each and every candidate source space partition, we can infer the most probable set of fMRI regions that has generated a given EEG scalp data window. We illustrate the method using EEG-correlated fMRI data acquired in a patient with ictal generalized spike-wave (GSW) discharges, to examine whether different networks are involved in the generation of the spike and the wave components, respectively. To this effect, we compared a family of 128 EEG source models, based on the combinations of seven regions haemodynamically involved (deactivated) during a prolonged ictal GSW discharge, namely: bilateral precuneus, bilateral medial frontal gyrus, bilateral middle temporal gyrus, and right cuneus. Bayesian model comparison has revealed the most likely model associated with the spike component to consist of a prefrontal region and bilateral temporal-parietal regions and the most likely model associated with the wave component to comprise the same temporal-parietal regions only. The result supports the hypothesis of different neurophysiological mechanisms underlying the generation of the spike versus wave components of GSW discharges. PMID- 19559799 TI - Perturbation of the BOLD response by a contrast agent and interpretation through a modified balloon model. AB - This study used an infusion of a paramagnetic contrast agent to perturb intravascular blood susceptibility and investigate its effect on the BOLD hemodynamic response. A three compartment BOLD signal model combined with a modified balloon model was developed to interpret the MR signal. This model incorporated arterial blood volume in order to simulate signal changes resulting from the contrast agent. The BOLD signal model was fitted to the experimental data to test the hypothesis that arterial blood volume changes during activation. It was found that allowing arterial blood volume to change, rather than assuming this change is negligible as often assumed in the literature, provides a better fit to the experimental data, particularly during the BOLD overshoot. The post stimulus undershoot was fitted well, regardless of whether the arterial blood volume was allowed to change, by assuming that this feature is due to delayed venous compliance. However the resultant elevation in post-stimulus blood volume decays with an extremely long time constant, taking more than 55 s to recover to baseline following a 4.8 s stimulus. The post-stimulus signal changes measured here could alternatively be described by a post-stimulus elevation in metabolism. An alternative model of oxygen extraction, in place of the Oxygen Limitation model, would be required to test this hypothesis. PMID- 19559800 TI - Lost in localization? The focus is meta-analysis. AB - The recent commentary by Derrfuss J, Mar RA. (2009). Lost in localization: the need for a universal coordinate database. Neuroimage, In Press proposed a universal coordinate database to archive functional neuroimaging results. In this response, we discuss our strategy in developing the BrainMap database, which was created as a mechanism to promote coordinate-based meta-analysis methods. PMID- 19559801 TI - A comparison between voxel-based cortical thickness and voxel-based morphometry in normal aging. AB - The morphology of cortical grey matter is commonly assessed using T1-weighted MRI together with automated computerised methods such as voxel-based morphometry (VBM) and cortical thickness measures. In the presented study we investigate how grey matter changes identified using voxel-based cortical thickness (VBCT) measures compare with local grey matter volume changes identified using VBM. We use data from a healthy aging population to perform the comparison, focusing on brain regions where age-related changes have been observed in previous studies. Our results show that overall, in healthy aging, VBCT and VBM yield very consistent results but VBCT provides a more sensitive measure of age-associated decline in grey matter compared with VBM. Our findings suggest that while VBCT selectively investigates cortical thickness, VBM provides a mixed measure of grey matter including cortical surface area or cortical folding, as well as cortical thickness. We therefore propose that used together, these techniques can separate the underlying grey matter changes, highlighting the utility of combining these complementary methods. PMID- 19559804 TI - Innate immune recognition of, and response to, Clostridium sordellii. AB - Clostridium sordellii, an anaerobic pathogen, has recently been associated with rapidly fatal infections following medically induced abortions and injecting drug use. Patients with C. sordellii infection display few signs of inflammation such as fever, or redness and pain at the site of infection. We hypothesized that this could be due to reduced recognition of the organism by Toll-like receptors (TLRs) of the innate immune system. An ELAM-NF-kappaB luciferase reporter system in TLR transfected HEK cells was used to measure TLR-dependent recognition of washed, heat-killed C. sordellii and other pathogenic clostridial species. Results demonstrated that all clostridia were well recognized by TLR2 alone and that responses were greatest when TLR2 was co-expressed with TLR6. Further, isolated human monocytes produced the pro-inflammatory cytokine TNFalpha and the immunoregulator IL-10 in response to C. sordellii. In addition, C. sordellii stimulated monocytes produced 30% less TNFalpha following treatment with an anti TLR2 blocking antibody. These data demonstrate that innate immune recognition of, and response to, cell-associated components of C. sordellii and other clostridial pathogens are mediated by TLR2 in combination with TLR6. We conclude that the characteristic absence of inflammatory signs and symptoms in C. sordellii infection is not related to inadequate immune detection of the organism, but rather is attributable to a species-specific immune system dysfunction that remains to be elucidated. PMID- 19559803 TI - Effects of different extracellular matrices and co-cultures on human limbal stem cell expansion in vitro. AB - To elucidate the effect of extracellular matrices (ECMs) and related and nonrelated-limbal feeder cells as substitutes for the in vivo niche on the phenotype and genotype of the limbal stem cell (SC) expansion in vitro, human limbal SCs were used. The limbus explants were expanded on human amniotic membrane (AM), commercial ECMs including matrigel (MAT), collagen (COL), and control (no ECM) in presence and absence of feeder cells including human limbal fibroblasts (LFs), a limbus-specific cell and mouse embryonic fibroblasts (MEFs). Proliferation, cell death, immunocytochemistry, expression of specific genes, ultrastructural characteristics, and size and granularity of expanded human limbal SCs in different groups were evaluated. The growth, cell proliferation, and survival of limbal SCs were enhanced by AM and MAT matrices. Ultrastructure and expression of stemness markers revealed that there was no significance difference between AM and MAT. However, flow cytometric analysis showed that the size and granularity of cultured cells increased in the presence of MAT and COL as well as in no ECM group. Moreover, co-culturing of limbal explants with LFs and MEFs on AM and MAT groups, enhanced the expansion and survival of cultured cells in comparison with others. In conclusion, the cultivation of human limbal explants on AM co-culturing with human LFs promises to be a good model for preparing undifferentiated epithelial sheets suitable for transplantation. PMID- 19559802 TI - Category-specific activations during word generation reflect experiential sensorimotor modalities. AB - According to the sensorimotor theory of lexicosemantic organization, semantic representations are neurally distributed and anatomically linked to category specific sensory areas. Previous functional neuroimaging studies have demonstrated category specificity in lexicosemantic representations. However, little evidence is available from word generation paradigms, which provide access to semantic representations while minimizing confounds resulting from low-level perceptual features of stimulus presentation. In this study, 13 healthy young adults underwent fMRI scanning while performing a word generation task, generating exemplars to nine different semantic categories. Each semantic category was assigned to one of three superordinate category types, based upon sensorimotor modalities (visual, motor, somatosensory) presumed to predominate in lexical acquisition. For word generation overall, robust activation was seen in left inferior frontal cortex. Analyses by sensorimotor modality categories yielded activations in brain regions related to perceptual and motor processing: Visual categories activated extrastriate cortex, motor categories activated the intraparietal sulcus and posterior middle temporal cortex, and somatosensory categories activated postcentral and inferior parietal regions. Our results are consistent with the sensorimotor theory, according to which lexicosemantic representations are distributed across brain regions participating in sensorimotor processing associated with the experiential components of lexicosemantic acquisition. PMID- 19559805 TI - Effects of feeding and hypoxia on cardiac performance and gastrointestinal blood flow during critical speed swimming in the sea bass Dicentrarchus labrax. AB - Previous studies have shown that if European sea bass are exercised after feeding, they can achieve a significantly higher maximum metabolic rate (MMR) than when fasted. They can meet combined metabolic demands of digestion (specific dynamic action, SDA) and maximal aerobic exercise, with no decline in swimming performance. If, however, exposed to mild hypoxia (50% saturation), bass no longer achieve higher MMR after feeding but they swim as well fed as fasted, due to an apparent ability to defer the SDA response. This study explored patterns of cardiac output (Q(A)) and blood flow to the gastrointestinal tract (Q(GI)) associated with the higher MMR after feeding, and with the ability to prioritise swimming in hypoxia. Sea bass (mean mass approximately 325 g, forklength approximately 27 cm) were instrumented with flow probes to measure Q(A) and Q(GI) during an incremental critical swimming speed (U(crit)) protocol in a tunnel respirometer, to compare each animal either fasted or 6h after a meal of fish fillet equal to 3% body mass. Feeding raised oxygen uptake (M(O2)) prior to exercise, an SDA response associated with increased Q(A) (+30%) and Q(GI) (+100%) compared to fasted values. As expected, when exercised the fed bass maintained the SDA load throughout the protocol and achieved 14% higher MMR than when fasted, and the same U(crit) (approximately 100 cm s(-1)). Both fed and fasted bass showed pronounced increases in Q(A) and decreases in Q(GI) during exercise and the higher MMR of fed bass was not associated with higher maximum Q(A) relative to when fasted, or to any differences in Q(GI) at maximum Q(A). In hypoxia prior to exercise, metabolic and cardiac responses to feeding were similar compared to normoxia. Hypoxia caused an almost 60% reduction to MMR and 30% reduction to U(crit), but neither of these traits differed between fed or fasted bass. Despite hypoxic limitations to MMR and U(crit), maximum Q(A) and patterns of Q(GI) during exercise in fasted and fed bass were similar to normoxia. Estimating GI oxygen supply from Q(GI) indicated that the ability of bass to prioritise aerobic exercise over SDA when metabolically limited by hypoxia was linked to an ability to defer elements of the SDA response occurring outside the GI tract. PMID- 19559807 TI - Brain gonadotropin releasing hormone3 expression variation during oogenesis and sexual behavior and its effect on pituitary hormonal expression in the blue gourami. AB - To gain a better insight regarding the roles of gonadotropin releasing hormone3 (GnRH3) in the regulation of reproduction in the suborder Labyrinthici, GnRH3 expression was investigated during the reproduction cycle of the male and female blue gourami (Trichogaster trichopterus). The full-length blue gourami brain GnRH3 gene was sequenced (EMBL acc. no. EU107388) and was found to be expressed in both the brain and pituitary of the blue gourami. High mRNA levels were detected only in the brain of females with oocytes in the maturation stage. Correspondingly, significantly greater mRNA levels of GnRH3 were detected in mature males than in immature ones. In primary cultures of dispersed pituitary cells, GnRH3 significantly increased betaFSH and betaLH subunit mRNA levels in cells from both females and males, whereas GH gene transcription was affected differently by GnRH3 in females, as compared to males. Thus, we propose that GnRH3 can act differentially in the blue gourami females and males. In females, GnRH3 may be involved in the final maturation stage of the oocyte and induces betaFSH, betaLH and GH gene expression; in males, it is engaged in sexual behavior and spermatogenesis regulation via betaFSH and betaLH stimulation and dowregulation of GH transcription. PMID- 19559806 TI - Adaptive response of brain tissue oxygenation to environmental hypoxia in non sedated, non-anesthetized arctic ground squirrels. AB - The present study examined the physiological mechanisms of the responses of brain tissue oxygen partial pressure (P(t)O(2)), brain temperature (T(brain)), global oxygen consumption (V(O2)), and respiratory frequency (f(R)) to hypoxia in non sedated and non-anesthetized arctic ground squirrels (Spermophilus parryii, AGS) and rats. We found that (1) in contrast to oxygen partial pressure in blood (P(a)O(2)), the baseline value of P(t)O(2) in summer euthermic AGS is significantly higher than in rats; (2) both P(t)O(2) and P(a)O(2) are dramatically reduced by inspired 8% O(2) in AGS and rats, but AGS have a greater capacity in P(t)O(2) to cope with environmental hypoxia; (3) metabolic rate before, during, and after hypoxic exposure is consistently lower in AGS than in rats; (4) the respiratory responding patterns to hypoxia in the two species differ in that f(R) decreases in AGS but increases in rats. These results suggest that (1) AGS have special mechanisms to maintain higher P(t)O(2) and lower P(a)O(2,) and these levels in AGS represent a typical pattern of adaptation of heterothermic species to and a brain protection from hypoxia; (2) AGS brain responds to hypoxia through greater decreases in P(t)O(2) and decreased f(R) and ventilation. In contrast, rat brain responds to hypoxia by less reduction in P(t)O(2) and increased f(R) and ventilation. PMID- 19559808 TI - The relationship between O2 chemoreceptors, cardio-respiratory reflex and hypoxia tolerance in the neotropical fish Hoplias lacerdae. AB - The localization, distribution and orientation of O(2) chemoreceptors associated with the control of cardio-respiratory responses were investigated in the neotropical, Hoplias lacerdae. Selective denervation of the cranial nerves (IX and X) was combined with chemical stimulation (NaCN) to characterize the gill O(2) chemoreceptors, and the fish were then exposed to gradual hypoxia to examine the extent of each cardio-respiratory response. Changes in heart rate (f(H)) and ventilation amplitude (V(amp)) were allied with chemoreceptors distributed on both internal and external surfaces of all gill arches, while ventilation rate (f) was allied to the O(2) chemoreceptors located only in the internal surface of the first gill arch. H. lacerdae exposed to gradual hypoxia produced a marked bradycardia (45%) and 50% increase in V(amp), but only a relatively small change in f (32%). Thus, the low f(R) response yet high V(amp) were in accord with the characterization of the O(2) chemoreceptors. Comparing these results from H. lacerdae with hypoxia-tolerant species revealed a relationship existent between general oxygenation of the individual species environment, its cardio-respiratory response to hypoxia and the characterization of O(2) chemoreceptors. PMID- 19559809 TI - Phytoestrogens and avian reproduction: Exploring the evolution and function of phytoestrogens and possible role of plant compounds in the breeding ecology of wild birds. AB - Phytoestrogens are secondary plant compounds, which can act to mimic estrogen and cause the disruption of estrogenic responses in organisms. Although there is a substantial body of research studying phytoestrogens, including their mechanisms of estrogenic effects, evolution, and detection in biological systems, little is known about their ecological significance. There is evidence, however, that an ecological relationship involving phytoestrogens exists between plants and animals-plants may produce phytoestrogens to reduce fecundity of organisms that eat them. Birds and other vertebrates may also exploit phytoestrogens to regulate their own reproduction-there are well known examples of phytoestrogens inhibiting reproduction in higher vertebrates, including birds. Also, common plant stressors (e.g., high temperature) increase the production of secondary plant compounds, and, as evidence suggests, also induce phytoestrogen biosynthesis. These observations are consistent with the single study ever done on phytoestrogens and reproduction in wild birds [Leopold, A.S., Erwin, M., Oh, J., Browning, B., 1976. Phytoestrogens adverse effects on reproduction in California quail. Science 191, 98-100.], which found that drought stress correlated with increased levels of phytoestrogens in plants, and that increased phytoestrogen levels correlated with decreased young. This review discusses the hypothesis that plants may have an effect on the reproduction of avian species by producing phytoestrogens as a plant defense against herbivory, and that birds may "use" changing levels of phytoestrogens in the vegetation to ensure that food resources will support potential young produced. Evidence from our laboratory and others appear to support this hypothesis. PMID- 19559810 TI - Multiple urea transporter proteins in the kidney of holocephalan elephant fish (Callorhinchus milii). AB - Reabsorption of filtered urea by the kidney is essential for retaining high levels of urea in marine cartilaginous fish. Our previous studies on the shark facilitative urea transporter (UT) suggest that additional UT(s) comprising the urea reabsorption system could exist in the cartilaginous fish kidney. Here, we isolated three cDNAs encoding UTs from the kidney of elephant fish, Callorhinchus milii, and termed them efUT-1, efUT-2 and efUT-3. efUT-1 is orthologous to known elasmobranch UTs, while efUT-2 and efUT-3 are novel UTs in cartilaginous fish. Two variants were found for efUT-1 and efUT-2, in which the NH(2)-terminal intracellular domain was distinct between the variants. Differences in potential phosphorylation sites were found in the variant-specific NH(2)-terminal domains. When expressed in Xenopus oocytes, all five UT transcripts including the efUT-1 and efUT-2 variants induced more than a 10-fold increase in [(14)C] urea uptake. Phloretin inhibited dose-dependently the increase of urea uptake, suggesting that the identified UTs are facilitative UTs. Molecular phylogenetic analysis revealed that efUT-1 and efUT-2 had diverged in the cartilaginous fish lineage, while efUT 3 is distinct from efUT-1 and efUT-2. The present finding of multiple UTs in elephant fish provides a key to understanding the molecular mechanisms of urea reabsorption system in the cartilaginous fish kidney. PMID- 19559811 TI - Comparison of microsomal prostaglandin E synthase-1 deletion and COX-2 inhibition in acute cardiac ischemia in mice. AB - The aim of the present study was to compare the effects of genetic mPGES-1 loss and COX-2 inhibition on myocardial damage after coronary occlusion. mPGES-1(-/-) mice and their wild-type littermates were injected with vehicle or COX-2 inhibitor (celecoxib), and 30min later the left coronary artery was surgically occluded. At 24h, myocardial infarct (MI) volume was measured histologically. Post-MI survival was reduced in WT mice receiving celecoxib (12/20) compared with vehicle-treated controls (12/12) or the loss of mPGES-1 (13/13) together with increased phosphokinase (CPK) and cardiac troponin-I release. Endogenous mPGES-1 expression was unchanged by ischemia in WT mice and absent in mPGES-1(-/-) hearts. COX-2 expression was markedly increased at 24h after MI in WT hearts; this upregulation was largely attenuated in mPGES-1(-/-) mice. We conclude that loss of mPGES-1 prevents the upregulation of COX-2 after myocardial infarct, and in contrast to inhibition of COX-2, does not increase ischemic myocardial damage. PMID- 19559812 TI - Functional identity of the active sites of crustacean and viral thymidylate synthases. AB - Thymidylate synthase (TS) catalyzes the synthesis of deoxythymidine monophosphate (dTMP), which is an essential precursor for DNA synthesis. The rationale underlying drug design is to identify compounds that differentially inhibit a viral or parasite enzyme vs. the host homologue. We studied the TS of the white spot syndrome virus (WSSV TS) and the corresponding TS from the host, the marine invertebrate shrimp Litopenaeus vannamei. TS is the only de novo source of dTMP and is essential for host and viral DNA replication. To establish proof of principle, we cloned a full-length TS cDNA from the white shrimp L. vannamei (shrimp TS) that corresponds to a deduced sequence of 289 amino acids and over expressed it to study inhibition of both shrimp and viral TSs. Steady-state kinetic parameters for both TSs are similar, and dissociation (K(d)) or half maximal inhibitory concentration constants (IC(50)) did not show differential inhibition between the folate analogues. Differences in their amino acid sequence are not reflected in theoretical molecular models of both TSs, since both appear to have identical active sites. These results suggest that the eukaryotic TS active site is very constrained into the functional residues involved in reductive methylation of 2'-deoxyuridine-5'-monophosphate (dUMP). PMID- 19559813 TI - Accumulation of cadmium in the otoliths and tissues of juvenile pink snapper (Pagrus auratus Forster) following dietary and waterborne exposure. AB - Laboratory experiments were conducted to examine if incorporation of Cd into the otoliths of juvenile pink snapper (Pagrus auratus Forster) was related to levels in the food or water. In the first experiment, fish were fed a regular diet (control group) or a Cd-contaminated diet (500mgCdkg(-1) or 1500mgCdkg(-1)) for 35days. In the second experiment, fish were exposed to waterborne Cd concentrations of <0.002microgL(-1) (control), 50microgL(-1), 100microgL(-1) and 150microgL(-1) for 35days. The sagittal otoliths were analysed using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Juvenile fish exposed to higher concentrations of waterborne or dietary Cd showed increased Cd levels in their otoliths. This study clearly demonstrated that both aqueous and dietary Cd exposures can result in Cd incorporation into the otoliths of pink snapper. PMID- 19559814 TI - Induction of estrogen-responsive gene transcription in the embryo, larval, juvenile and adult life stages of zebrafish as biomarkers of short-term exposure to endocrine disrupting chemicals. AB - Real-time quantitative RT-PCR was adopted to investigate the transcription of the estrogen-responsive genes in different developmental stages of zebrafish exposed to endocrine disrupting chemicals (EDCs). The lowest observed effect concentrations (LOECs) of 17beta-estradiol for inducing vtg1 transcription were 0.25, 0.5, 0.25 and 14 days), and short length of stay (< 6 days and alive). Candidate predictor variables were selected based on their availability in versions 2.35, 2.41, and 2.52.1 of the STS NCD and their presence in (or ability to be mapped to) version 2.61. Potential predictor variables were screened for overall prevalence in the study population, missing data frequency, coding concerns, bivariate relationships with outcomes, and their presence in previous STS or other CABG risk models. Supervised backwards selection was then performed with input from an expert panel of cardiac surgeons and biostatisticians. After successfully validating the fit of the models, the development and validation samples were subsequently combined, and the final regression coefficients were estimated using the overall combined (development plus validation) sample. RESULTS: The c-index for the mortality model was 0.812, and the c-indices for other endpoints ranged from 0.653 for reoperation to 0.793 for renal failure in the validation sample. Plots of observed versus predicted event rates revealed acceptable calibration in the overall population and in numerous subgroups. When patients were grouped into categories of predicted risk, the absolute difference between the observed and expected event rates was less than 1.5% for each endpoint. The final model intercept and coefficients are provided. CONCLUSIONS: New STS risk models have been developed for CABG mortality and eight other endpoints. Detailed descriptions of model development and testing are provided, together with the final algorithm. Overall model performance is excellent. PMID- 19559824 TI - The Society of Thoracic Surgeons 2008 cardiac surgery risk models: part 3--valve plus coronary artery bypass grafting surgery. AB - BACKGROUND: Since 1999, The Society of Thoracic Surgeons (STS) has published two risk models that can be used to adjust the results of valve surgery combined with coronary artery bypass graft surgery (CABG). The most recent was developed from data for patients who had surgery between 1994 and 1997 using operative mortality as the only endpoint. Furthermore, this model did not specifically consider mitral valve repair plus CABG, an increasingly common procedure. Consistent with STS policy of periodically updating and improving its risk models, new models for valve surgery combined with CABG have been developed. These models specifically address both perioperative morbidity and mitral valve repair, and they are based on contemporary data. METHODS: The final study population consisted of 101,661 procedures, including aortic valve replacement (AVR) plus CABG, mitral valve replacement (MVR) plus CABG, or mitral valve repair (MVRepair) plus CABG between January 1, 2002, and December 31, 2006. Model outcomes included operative mortality, stroke, deep sternal wound infection, reoperation, prolonged ventilation, renal failure, composite major morbidity or mortality, prolonged postoperative length of stay, and short postoperative length of stay. Candidate variables were screened for frequency of missing data, and imputation techniques were used where appropriate. Stepwise variable selection was employed, supplemented by advice from an expert panel of cardiac surgeons and biostatisticians. Several variables were forced into models to insure face validity (eg, atrial fibrillation for the permanent stroke model, sex for all models). Based on preliminary analyses of the data, a single model was employed for valve plus CABG, with indicator variables for the specific type of procedure. Interaction terms were included to allow for differential impact of predictor variables depending on procedure type. After validating the model in the 40% validation sample, the development and validation samples were then combined, and the final model coefficients were estimated using the overall 100% combined sample. The final logistic regression model was estimated using generalized estimating equations to account for clustering of patients within institutions. RESULTS: The c-index for mortality prediction for the overall valve plus CABG population was 0.75. Morbidity model c-indices for specific complications (permanent stroke, renal failure, prolonged ventilation > 24 hours, deep sternal wound infection, reoperation for any reason, major morbidity or mortality composite, and prolonged postoperative length of stay) for the overall group of valve plus CABG procedures ranged from 0.622 to 0.724, and calibration was excellent. CONCLUSIONS: New STS risk models have been developed for heart valve surgery combined with CABG. These are the first valve plus CABG models that also include risk prediction for individual major morbidities, composite major morbidity or mortality, and short and prolonged length of stay. PMID- 19559823 TI - The Society of Thoracic Surgeons 2008 cardiac surgery risk models: part 2- isolated valve surgery. AB - BACKGROUND: Adjustment for case-mix is essential when using observational data to compare surgical techniques or providers. That is most often accomplished through the use of risk models that account for preoperative patient factors that may impact outcomes. The Society of Thoracic Surgeons (STS) uses such risk models to create risk-adjusted performance reports for participants in the STS National Adult Cardiac Surgery Database (NCD). Although risk models were initially developed for coronary artery bypass surgery, similar models have now been developed for use with heart valve surgery, particularly as the proportion of such procedures has increased. The last published STS model for isolated valve surgery was based on data from 1994 to 1997 and did not include patients undergoing mitral valve repair. STS has developed new valve surgery models using contemporary data that include both valve repair as well as replacement. Expanding upon existing valve models, the new STS models include several nonfatal complications in addition to mortality. METHODS: Using STS data from 2002 to 2006, isolated valve surgery risk models were developed for operative mortality, permanent stroke, renal failure, prolonged ventilation (> 24 hours), deep sternal wound infection, reoperation for any reason, a major morbidity or mortality composite endpoint, prolonged postoperative length of stay, and short postoperative length of stay. The study population consisted of adult patients who underwent one of three types of valve surgery: isolated aortic valve replacement (n = 67,292), isolated mitral valve replacement (n = 21,229), or isolated mitral valve repair (n = 21,238). The population was divided into a 60% development sample and a 40% validation sample. After an initial empirical investigation, the three surgery groups were combined into a single logistic regression model with numerous interactions to allow the covariate effects to differ across these groups. Variables were selected based on a combination of automated stepwise selection and expert panel review. RESULTS: Unadjusted operative mortality (in-hospital regardless of timing, and 30-day regardless of venue) for all isolated valve procedures was 3.4%, and unadjusted in-hospital morbidity rates ranged from 0.3% for deep sternal wound infection to 11.8% for prolonged ventilation. The number of predictors in each model ranged from 10 covariates in the sternal infection model to 24 covariates in the composite mortality plus morbidity model. Discrimination as measured by the c-index ranged from 0.639 for reoperation to 0.799 for mortality. When patients in the validation sample were grouped into 10 categories based on deciles of predicted risk, the average absolute difference between observed versus predicted events within these groups ranged from 0.06% for deep sternal wound infection to 1.06% for prolonged postoperative stay. CONCLUSIONS: The new STS risk models for valve surgery include mitral valve repair as well as multiple endpoints other than mortality. Model coefficients are provided and an online risk calculator is publicly available from The Society of Thoracic Surgeons website. PMID- 19559826 TI - [Aggressive and violent behavior in adolescents]. PMID- 19559827 TI - [Managing the suicidal crisis]. PMID- 19559828 TI - [Indications for crenotherapy]. PMID- 19559829 TI - [Electroconvulsive therapy today]. PMID- 19559830 TI - Search for premalignant mucosal lesions: does endoscopic measurement of oxygen saturation by differential path-length spectroscopy help? PMID- 19559831 TI - In praise of folly: the Erasmus score for intestinal metaplasia. PMID- 19559833 TI - ERCP complication rates: how low can we go? PMID- 19559832 TI - A comparison of the Niti-D biliary uncovered stent and the uncovered Wallstent in malignant biliary obstruction. AB - BACKGROUND: The conformability of uncovered self-expandable metal stents (SEMSs) plays an important role in maintaining stent patency. However, whether increased conformability can prolong the duration of SEMS patency remains to be proved. OBJECTIVE: The aim of this study was to examine the efficacy and complication rates of the Niti-D biliary uncovered metal stent (NDS), which is more conformable than the uncovered Wallstent. DESIGN: Nonrandomized, retrospective study. SETTING: Tertiary-care academic medical center. PATIENTS: From March 2005 to July 2007, 101 patients received an NDS (41 cases) or a Wallstent (60 cases) for malignant biliary obstruction. INTERVENTIONS: SEMS placement. RESULTS: Stent occlusion occurred in 11 patients (26.8%) with the NDS and 17 patients (28.3%) with the Wallstent. The median duration of stent patency tended to be longer for the NDS group (153 days) than for the Wallstent group (124 days); however, the difference was not statistically significant (P = .204). The median duration of overall survival of patients was 160 days for the NDS and 148 days for the Wallstent. The subgroup analysis showed that 27 patients had hilar obstruction (NDS 13, Wallstent 14). The median duration of stent patency was 249 days for the NDS group and 76 days for the Wallstent group; this difference was statistically significant (P = .006). The complications included pancreatitis in 3 NDS cases and 5 Wallstent cases. LIMITATION: The absence of prospective randomized recruitment. CONCLUSION: The results of this study showed no significant differences between the NDS and the Wallstent for the palliative endoscopic management of malignant biliary obstruction. There were no significant differences in patency, complication rates, and patient survival between the more conformable NDS and the conventional Wallstent. However, the NDS, which has good conformability, may be preferred for hilar obstruction. PMID- 19559834 TI - Customization and the cost of eliminating disparities in colon cancer. PMID- 19559835 TI - Parenteral nutrition 102: Complications, monitoring, and home use. PMID- 19559836 TI - Ergonomics and GI endoscopy. PMID- 19559837 TI - Management of transmesenteric tunnel jejunal strictures with endoscopic dilation by using achalasia balloons (with videos). AB - BACKGROUND: Roux-en-Y gastric bypass is the most commonly performed bariatric operation in the United States. Transmesenteric tunnel (TMT) or mesocolic jejunal stricture is an unusual postoperative complication that requires another operation. We hypothesize that endoscopic dilation by using achalasia dilatation balloons can be used to treat some TMT jejunal strictures. PATIENTS: This study involved 6 consecutive cases of TMT stricture. INTERVENTION: The TMT strictures were dilated by using achalasia balloons (30-40 mm) under fluoroscopic and endoscopic guidance. RESULTS: With endotherapy, 4 patients with late onset of symptoms (>3 weeks after Roux-en-Y gastric bypass) have not required another operation to date, with a follow-up of at least 5 to 12 months. Two patients with early onset of symptoms (<3 weeks after surgery) required operations, and 1 of these patients (symptoms onset <7 days after surgery) developed jejunal perforation within the stricture during dilation due to underlying jejunal ischemia within the stricture. LIMITATIONS: Small case number and limited follow up period. CONCLUSION: Endoscopic dilation by using achalasia balloons can be used to treat some TMT jejunal strictures without another operation. Surgery should be considered in patients with early onset of obstructive symptoms and/or with jejunal ischemia within the stricture. PMID- 19559838 TI - Colonoscopic full-thickness resection of the colon in a porcine model as a prelude to endoscopic surgery of difficult colon polyps: a novel technique (with videos). AB - BACKGROUND: Colonoscopic full-thickness resection (CFTR) of the colon may obviate the need for surgical resection of benign lesions. OBJECTIVE: To develop an animal model for CFTR of the colon followed by endoscopic suture closure with through-the-endoscope devices. DESIGN: Pilot study. SETTING: University medical center. ANIMALS: Twenty pigs. INTERVENTIONS: A 2-cm circular area was resected on the antimesenteric side of the colon (phase 1, n = 10) and on the mesenteric side (phase 2, n = 10) by using an insulated tip knife cut followed by the use of a grasping forceps and a snare to resect and retrieve the specimen. The tissue apposition system was used to close the defect. MAIN OUTCOME MEASUREMENTS: Resection and closure times were recorded. The animals were euthanized at 2 weeks and examined for peritonitis, adhesions, wound healing, and T-tag injury to adjacent viscera. RESULTS: The CFTR was successful in all 20 attempts. The median resection time was 6 minutes (range 2.5-35 minutes). Suture closure was successful in 19 animals. It took a median time of 41 minutes (range 21-125 minutes) and 4 sutures to close the defect. Eighteen animals survived without clinical signs of distress; there was a well-healed scar without peritonitis or distant adhesions on necropsy at 2 weeks. One animal failed to thrive, and necropsy revealed mild peritonitis, small abscesses, distant adhesions, and a 2 mm hole at the suture site. Two of the 132 T-tags were inserted in the adjacent viscera. LIMITATIONS: Colon resection in the proximal colon was not studied. CONCLUSIONS: In this animal model, CFTR of the colon followed by suture closure can be accomplished successfully by using through-the-endoscope devices. PMID- 19559839 TI - Ingestion of an unusual esophageal foreign body: what time is it? PMID- 19559840 TI - Isolated gastric mucosal hemorrhages in idiopathic thrombocytopenic purpura. PMID- 19559841 TI - An unusual cause of diarrhea: diffuse intestinal nodular lymphoid hyperplasia in association with selective immunoglobulin A deficiency (with video). PMID- 19559842 TI - Good stents gone bad: endoscopic treatment of proximally migrated pancreatic duct stents. AB - BACKGROUND: Pancreatic duct stents are used for a variety of endoscopic pancreatic manipulations, and small surgical stents are used prophylactically to bridge pancreatic-enteric anastomoses. With increasing use of pancreatic stents, many complications have been recognized. OBJECTIVE: To determine the complications and outcomes of pancreatic stent migration. DESIGN: Case series from a retrospective review of all cases of upstream or proximally migrated pancreatic duct stents, placed either endoscopically or surgically, identified between 2000 and 2007. SETTING: Tertiary referral center. PATIENTS: This study involved 33 patients; 23 postendoscopic and 10 postsurgical stents. MAIN OUTCOME MEASUREMENTS: Retrieval rates, minor/major complications. RESULTS: Endoscopic stents had a successful retrieval rate of 78%. Most patients were asymptomatic. The most common procedure was balloon extraction (8 of 18; 44%). Nine patients required multiple procedures (3 patients, 2 attempts; 5 patients, 3 attempts; 1 patient, 4 attempts). Five stents could not be retrieved. Of these, 4 were associated with downstream stenosis. Four patients underwent surgery, and 1 patient was treated with observation. Complications included pancreatic duct disruption (1 of 23), stent fragmentation (1 of 23), and postprocedure pancreatitis (1 of 23). Surgically placed stents had a successful retrieval rate of 80%. Most surgical stents had migrated into the biliary tree (8 of 10). All of these patients were symptomatic with pain or fever. Two stents could not be retrieved; 1 of those patients underwent surgery. LIMITATION: Retrospective study. CONCLUSION: The majority of upstream-migrated stents can be endoscopically removed. Despite manipulation of the pancreatic duct, pancreatitis was infrequent. Surgically placed pancreatic stents migrate downstream and into the open biliary anastomosis and are associated with pain, cholangitis, or liver abscesses. PMID- 19559844 TI - Pharmacotherapy for gastric antral vascular ectasia: dramatic response to tranexamic acid. PMID- 19559845 TI - Could we standardize the injection technique and regimen of N-butyl-2 cyanoacrylate injection for the treatment of gastric fundal varices? PMID- 19559847 TI - Transanal endoscopic microsurgical platform for natural orifice surgery. PMID- 19559849 TI - PillCam colon capsule endoscopy: the patient's perspective. PMID- 19559850 TI - Another cause of "segmental" inflammation in ulcerative colitis. PMID- 19559853 TI - Novel prediction of interactive mode between antibiotics and their DNA/protein targets based on the antibiotic structure parameters. AB - This paper focuses on investigating the structure parameters of antibiotics which have decisive influence on the interactive mode between antibiotics and DNAs. The analytical results of Best Prediction Set Support Vector Machine Method (BPSSVM) show that the number of N atoms, the number of double bonds, the number of lone pair electrons, the molar refractivity and the molecular radius are the main structure parameters which functionally decide their reactive modes. It is clear that the BPSSVM is potent in predicting the interactive modes between antibiotics and DNAs. PMID- 19559852 TI - Sample treatment and determination of pesticide residues in fatty vegetable matrices: a review. AB - A demanding task in pesticide residue analysis is yet the development of multi residue methods for the determination of pesticides in vegetables with relatively high fat content (i.e. edible oils and fatty vegetables). The separation of pesticides and other chemical contaminants from high-fat food samples prior to subsequent steps in the analytical process is yet a challenging issue to which much effort in method development has being applied. This review addresses the main sample treatment methodologies for pesticide residue analysis in fatty vegetable matrices. Even with the advent of advanced hyphenated techniques based on mass spectrometry these complex fatty matrices usually require extensive sample extraction and purification. Current methods involve the use of one or the combination of some of the following techniques for both the sample extraction and clean-up steps: liquid-liquid partitioning, solid-phase extraction (SPE), gel permeation chromatography (GPC), matrix solid-phase dispersion (MSPD), etc. An overview of methods developed for these contaminants in fatty vegetables matrices is presented. Sample extraction and purification techniques are discussed and their most recent applications are highlighted. This review emphasizes that sample preparation is a critical step, but also the determination method is, and cannot be treated separately from sample treatment. In recent years, the appearance and use of new, more polar pesticides has fostered the development of liquid chromatography/mass spectrometry (LC-MS) besides gas chromatography. The main features of LC-MS for the analysis of multi-class pesticides in fatty vegetable samples will be also underlined, with an emphasis on the multi-class, multi-residue strategy and the difficulties associated. PMID- 19559854 TI - Investigation on enhanced chemiluminescence reaction systems with bis(hydrogenperiodato) argentate(III) complex anion for fluoroquinolones synthetic antibiotics. AB - A novel chemiluminescence (CL) reaction system with bis(hydrogenperiodato) argentate(III) complex anion (Ag(III) complex, [Ag(HIO(6))(2)](5-)), for the first time, is developed for the determination of lomefloxacin (LMFX), enrofloxacin (ENLX) and pefloxacin (PFLX). The possible CL emission mechanism was discussed by comparing the fluorescence emission with CL spectra. The CL conditions of [Ag(HIO(6))(2)](5-)-H(2)SO(4)-LMFX/ENLX/PFLX systems were investigated and optimized. Under the optimized experimental conditions, the CL intensity is proportional to the concentration of the drugs in the range 0.2994 36.80x10(-7)g mL(-1) for LMFX, 4.00-30.0x10(-7)g mL(-1) for ENLX and 1.54 27.64x10(-7)g mL(-1) for PFLX. The limit of detection (s/n=3) was 9.1x10(-9)g mL( 1) for LMFX, 3.1x10(-9)g mL(-1) for ENLX and 4.4x10(-9)g mL(-1) for PFLX. The recovery of LMFX, ENLX and PELX from the spiked pharmaceutical preparations was in the range of 92.3-105% with the RSDs of 0.5-2.7%. For urine, serum and milk samples the recoveries of the three drugs were in the range of 85.1-107% for LMFX with the RSDs of 2.3-3.4%. 80.2-112% for ENLX with the RSDs of 1.4-2.8%, and 87.8 114% for PFLX with the RSDs of 1.6-2.7%. The proposed method was applied successfully to the determination of these compounds in real samples. PMID- 19559855 TI - Grafting of molecularly imprinted polymers from the surface of silica gel particles via reversible addition-fragmentation chain transfer polymerization: a selective sorbent for theophylline. AB - Molecularly imprinted polymers (MIPs) were grafted successfully from the surface of silica gel particles via surface initiated reversible addition-fragmentation chain transfer (RAFT) polymerization using RAFT agent functionalized silica gel as the chain transfer agent. The intrinsic characteristics of the controlled/living polymerization mechanism of RAFT allowed for the effective control of the grafting process. Thus the grafting copolymerization of methacrylic acid and divinyl benzene in the presence of template theophylline led to thin MIP film coating silica gel (MIP-Silica). The thickness of MIP film prepared in this study is about 1.98 nm, which was calculated from the nitrogen sorption analysis results. Measured binding kinetics for theophylline to the MIP Silica and MIPs prepared by conventional bulk polymerization demonstrated that MIP-Silica had improved mass-transfer properties. In addition, the theophylline imprinted MIP-Silica was used as the sorbent in solid-phase extraction to determine theophylline in blood serum with satisfactory recovery higher than 90%. Nonspecific adsorption of interfering compounds can be eliminated by a simple elution with acetonitrile, without sacrificing the selective binding of theophylline. PMID- 19559856 TI - A solid phase extraction using a chelate resin immobilizing carboxymethylated pentaethylenehexamine for separation and preconcentration of trace elements in water samples. AB - A chelate resin immobilizing carboxymethylated pentaethylenehexamine (CM-PEHA resin) was prepared, and the potential for the separation and preconcentration of trace elements in water samples was evaluated through the adsorption/elution test for 62 elements. The CM-PEHA resin could quantitatively recover various elements, including Ag, Cd, Co, Cu, Fe, Ni, Pb, Ti, U, and Zn, and rare earth elements over a wide pH range, and also Mn at pH above 5 and V and Mo at pH below 7. This resin could also effectively remove major elements, such as alkali and alkaline earth elements, under acidic and neutral conditions. Solid phase extraction using the CM-PEHA resin was applicable to the determination of 10 trace elements, Cd, Co, Cu, Fe, Mn, Mo, Ni, Pb, V, and Zn, in certified reference materials (EnviroMAT EU L-1 wastewater and ES-L-1 ground water) and treated wastewater and all elements except for Mn in surface seawater using inductively coupled plasma atomic emission spectrometry. The detection limits, defined as 3 times the standard deviation for the procedural blank using 500 mL of purified water (50-fold preconcentration, n=8), ranged from 0.003 microg L(-1) (Mn) to 0.28 microg L(-1) (Zn) as the concentration in 500 mL of solution. PMID- 19559857 TI - Detection of deletion mutations in DNA using water-soluble cationic fluorescent thiophene copolymer. AB - In this contribution, we designed a fluorescent thiophene copolymer to detect insertion/deletion mutation in DNA by doping aldehyde group in the main chain. The fluorescence of the copolymer could be dramatically quenched on the addition of single-stranded DNA (ssDNA) via strong electrostatic interactions and electronic/energy transfer. Although the complementary ssDNA made the fluorescence recover, the hydrogen bonds and chemical coupling also played a significant role between the unpaired bases and aldehyde group, which could differentiate the subtle differences in such mutant DNA. The influence of buffer pH, concentration of NaCl, heating time and the temperature was systemically investigated and the proposed method was then successfully applied to detect real sample. With the respect to the linearity, limit of detection precision, specificity, this procedure could provide sensitive methodologies for the rapid detection and identification of nucleic acids. PMID- 19559858 TI - Interdigitated array microelectrode based impedance immunosensor for detection of avian influenza virus H5N1. AB - Continuous outbreaks of avian influenza (AI) in recent years with increasing threat to animals and human health have warranted the urgent need for rapid detection of pathogenic AI viruses. In this study, an impedance immunosensor based on an interdigitated array (IDA) microelectrode was developed as a new application for sensitive, specific and rapid detection of avian influenza virus H5N1. Polyclonal antibodies against AI virus H5N1 surface antigen HA (Hemagglutinin) were oriented on the gold microelectrode surface through protein A. Target H5N1 viruses were then captured by the immobilized antibody, resulting in a change in the impedance of the IDA microelectrode surface. Red blood cells (RBCs) were used as biolabels for further amplification of the binding reaction of the antibody-antigen (virus). The binding of target AI H5N1 onto the antibody modified IDA microelectrode surface was further confirmed by atomic force microscopy. The impedance immunosensor could detect the target AI H5N1 virus at a titer higher than 10(3)EID(50)/ml (EID(50): 50% Egg Infective Dose) within 2h. The response of the antibody-antigen (virus) interaction was shown to be virus titer-dependent, and a linear range for the titer of H5N1 virus was found between 10(3) and 10(7)EID(50)/ml. Equivalent circuit analysis indicated that the electron transfer resistance of the redox probe [Fe(CN)(6)](3-/4-) and the double layer capacitance were responsible for the impedance change due to the protein A modification, antibody immobilization, BSA (bovine serum albumin) blocking, H5N1 viruses binding and RBCs amplification. No significant interference was observed from non-target RNA viruses such as Newcastle disease virus and Infectious Bronchitis disease virus. (The H5N1 used in the study was inactivated virus.). PMID- 19559860 TI - High-sensitivity detection of polysaccharide using phosphodiesters quaternary ammonium salt as probe by decreased resonance light scattering. AB - Phosphodiesters quaternary ammonium salt (PQAS) displayed quite intense light scattering in aqueous solution under the optimum condition. In addition, the resonance light scattering (RLS) signal of PQAS was remarkably decreased after adding trace amount polysaccharide with the maximum peak located at 391 nm. It was found that the decreased RLS intensity of the PQAS-PPGL system (DeltaI(RLS)) was in proportion to PPGL concentration in the range of 0.1-30 ng mL(-1), with a lower detection limit of 0.05 ng mL(-1). Based on this rare decreased RLS phenomenon, the novel method of the determination of purified polysaccharide of Gracilaria Lemaneiformis (PPGL) at nanogram level was proposed in this contribution. The proposed approach was used to determine purified polysaccharide extracted from Gracilaria Lemaneiformis with satisfactory results. Compared with the reported polysaccharide assays, this proposed method has good selectivity, high sensitivity and is especially simple and convenient. Moreover, the mechanism of the reaction between PQAS and polysaccharide was investigated by RLS, fluorescence, and fluorescence lifetime spectra. PMID- 19559859 TI - Electrochemiluminescence from tris(2,2'-bipyridyl)ruthenium(II)-graphene-Nafion modified electrode. AB - An electrochemiluminescence (ECL) sensor based on Ru(bpy)(3)(2+)-graphene-Nafion composite film was developed. The graphene sheet was produced by chemical conversion of graphite, and was characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), and Raman spectroscopy. The introduction of conductive graphene into Nafion not only greatly facilitates the electron transfer of Ru(bpy)(3)(2+), but also dramatically improves the long-term stability of the sensor by inhibiting the migration of Ru(bpy)(3)(2+) into the electrochemically inactive hydrophobic region of Nafion. The ECL sensor gives a good linear range over 1x10(-7) to 1x10(-4)M with a detection limit of 50 nM towards the determination of tripropylamine (TPA), comparable to that obtained by Nafion-CNT. The ECL sensor keeps over 80% and 85% activity towards 0.1 mM TPA after being stored in air and in 0.1M pH 7.5 phosphate buffer solution (PBS) for a month, respectively. The long-term stability of the modified electrode is better than electrodes modified with Nafion, Nafion-silica, Nafion-titania, or sol-gel films containing Ru(bpy)(3)(2+). Furthermore, the ECL sensor was successfully applied to the selective and sensitive determination of oxalate in urine samples. PMID- 19559861 TI - Admicelle-enhanced synchronous fluorescence spectrometry for the selective determination of polycyclic aromatic hydrocarbons in water. AB - A simple and rapid method for the highly sensitive determination of polycyclic aromatic hydrocarbons (PAHs) in water was developed. Benzo[a]pyrene, benzo[k]fluoranthene, perylene, and pyrene in water were concentrated into sodium dodecyl sulfate (SDS)-alumina admicelles. The collection was performed by adding SDS and alumina particles into the sample solution at pH 2. After gentle mixing, the resulting suspension was passed through a membrane filter to collect the SDS admicelles containing highly concentrated PAHs. The filter was placed on a slide glass and then covered admicellar layer with a fused silica glass plate before setting in a fluorescence spectrometer. Benzo[a]pyrene, benzo[k]fluoranthene, perylene, and pyrene were selectively determined by the synchronous fluorescence scan (SFS) analysis with keeping wavelength intervals between excitation and emission to 98, 35, 29, and 45 nm, respectively. Because of the minimum spectral overlapping, 1-40 ng l(-1) of benzo[a]pyrene, benzo[k]fluoranthene, and perylene as well as 10-150 ng l(-1) of pyrene were selectively determined with eliminating the interferences of other 12 PAHs. The detection limits were 0.3 ng l(-1) for benzo[a]pyrene, benzo[k]fluoranthene, and perylene, and 1 ng l(-1) for pyrene. They were 2-3 orders of magnitude lower than the detection limits in normal aqueous micellar solutions. The application to water analysis was studied. PMID- 19559862 TI - Lead determination in whole blood by laser ablation coupled with inductively coupled plasma mass spectrometry. AB - This work describes a simple procedure for blood lead level determination. The proposed method requires little sample pretreatment and subsequent direct analysis of a dried blood spot on a filter membrane using laser ablation coupled with inductively coupled plasma mass spectrometry (LA-ICP-MS). In general, LA-ICP MS studies are somewhat limited by the lack of matrix-matched standards for calibration purposes. Here we describe aqueous standard calibration and matrix matched calibration methods. This method was validated by analysis of the reference materials. With the matrix-matched calibration method, the recovery ranged from 97.8% to 112.8%, while the aqueous standard calibration method ranged 90.4% to 122.4%. The lower detection limit was estimated as 0.1 ng mL(-1). The determination precision, expressed as the relative standard deviation (RSD), was not worse than 10% for all results. A sample throughput of approximately 5 min per sample made it possible to rapidly screen a large number of samples. PMID- 19559863 TI - Determination of Prometryne in water and soil by HPLC-UV using cloud-point extraction. AB - A CPE-HPLC (UV) method has been developed for the determination of Prometryne. In this method, non-ionic surfactant Triton X-114 was first used to extract and pre concentrate Prometryne from water and soil samples. The separation and determination of Prometryne were then carried out in an HPLC-UV system with isocratic elution using a detector set at 254 nm wavelength. The parameters and variables that affected the extraction were also investigated and the optimal conditions were found to be 0.5% of Triton X-114 (w/v), 3% of NaCl (w/v) and heat assisted at 50 degrees C for 30 min. Using these conditions, the recovery rates of Prometryne ranged from 92.84% to 99.23% in water and 85.48% to 93.67% in soil, respectively, with all the relative standard deviations less than 3.05%. Limit of detection (LOD) and limit of quantification (LOQ) were 3.5 microg L(-1) and 11.0 microg L(-1) in water and 4.0 microg kg(-1) and 13.0 microg kg(-1) in soil, respectively. Thus, we developed a method that has proven to be an efficient, green, rapid and inexpensive approach for extraction and determination of Prometryne from soil samples. PMID- 19559864 TI - DNA hybridization electrochemical biosensor using a functionalized polythiophene. AB - A simple and label-free electrochemical sensor for recognition of the DNA sensor event was prepared by electrochemical polymerization of 4-hydroxyphenyl thiophene 3-carboxylate. Poly(4-hydroxyphenyl thiophene-3-carboxylate) (PHPT) was synthesized electrochemically onto glassy carbon electrode and characterized by cyclic voltammetry, FTIR and AFM measurements. An ODN-probe was physisorbed onto PHPT film and tested on hybridization with complementary ODN segments. A biological recognition can be monitored by comparison with electrochemical signal (cyclic voltammogram) of single and double strand state oligonucleotide. The oxidation current of double strand state oligonucleotide is lower than that of single strand, that is corresponding to the decrease of electroactivity of PHPT with the increase of stiffness of polymer structure. Physisorbed ODN-probe and its hybridization were observed morphologically onto ITO electrodes using AFM. The sensitivity of the electrochemical sensor is 0.02 microA/nmol, detection limit is 1.49 nmol and it has good selectivity. PMID- 19559865 TI - Detection of polycyclic aromatic hydrocarbon (PAH) compounds in artificial sea water using surface-enhanced Raman scattering (SERS). AB - This paper reports an accurate synthesis of surface-enhanced Raman scattering (SERS) active substrates, based on gold colloidal monolayer, suitable for in situ environmental analysis. Quartz substrates were functionalized by silanization with (3-mercaptopropyl)trimethoxysilane (MPMS) or (3-aminopropyl)trimethoxysilane (APTMS) and they subsequently reacted with colloidal suspension of gold metal nanoparticles: respectively, the functional groups SH and NH(2) bound gold nanoparticles. Gold nanoparticles were prepared by the chemical reduction of HAuCl(4) using sodium tricitrate and immobilized onto silanized quartz substrates. Active substrate surface morphology was characterized with scanning electron microscopy (SEM) measurements and gold nanoparticles presented a diameter in the range 40-100 nm. Colloidal hydrophobic films, allowing nonpolar molecule pre-concentration, were obtained. The surfaces exhibit strong enhancement of Raman scattering from molecules adsorbed on the films. Spectra were recorded for two PAHs, naphthalene and pyrene, in artificial sea-water (ASW) with limits of detection (LODs) of 10 ppb for both on MPMS silanized substrates. PMID- 19559866 TI - A single standard calibration module for flow analysis systems based on solenoid microdevices. AB - Only two computer-controlled microsolenoid devices, namely two micropumps or one micropump and one microvalve, are sufficient for the construction of on-line dilution modules useful in several flow analytical systems for the calibration using single standard. Three simple constructions of such modules were tested and compared. The most promising is the one based on the concept of a microvalve controlling dilution ratio of the standard and a solenoid micropump playing a double role: solution pumping device and mixing segments homogenizer. All investigated modules were tested with paired emitter detector diode (PEDD) as photometric flow-through detector and bromothymol blue as a model analyte. The best module was implemented into more advanced flow-injection system dedicated for optical detection of alkaline phosphatase activity using UV-PEDD-based flow through detector for the enzyme reaction product. PMID- 19559867 TI - Sugar determination via the homogeneous reduction of Au salts: a novel optical measurement. AB - A novel optical assay for sugar determination based on the catalytic and biocatalytic growth of gold nanoparticles (AuNPs) is presented. The reaction of carbohydrates with these Au salts in alkaline media generates AuNPs at room temperature (RT) without the need for Au seeds in the solution or fibrous mesh. The optical properties of the resulting AuNPs relates to the total reducing sugar content of the samples analyzed. The development of such inexpensive optical assay was evaluated qualitatively and quantitatively on food beverages and honey samples. Its application can be of help to control the glucose content of the diet or easily extended in a host of industrial, biomedical and clinical fields. PMID- 19559868 TI - Determination of indole-3-acetic acid and indole-3-butyric acid in mung bean sprouts using high performance liquid chromatography with immobilized Ru(bpy)3(2+)-KMnO4 chemiluminescence detection. AB - A novel method for determination of indole-3-acetic acid (IAA) and indole-3 butyric acid (IBA) in an extract from mung bean sprouts using high performance liquid chromatography (HPLC) with chemiluminescence (CL) detection is described. The method is based on the CL reaction of auxin (indole-3-acetic acid and indole 3-butyric acid) with acidic potassium permanganate (KMnO(4)) and tris(2,2' bipyridyl)ruthenium(II), which was immobilized on the cationic ion-exchange resin. The chromatographic separation was performed on a Nucleosil RP-C18 column (i.d.: 250 mm x 4.6 mm, particle size: 5 microm, pore size: 100) with an isocratic mobile phase consisting of methanol-water-acetic acid (45:55:1, v/v/v). At a flow rate of 1.0 mL min(-1), the total run time was 20 min. Under the optimal conditions, the linear ranges were 5.0x10(-8) to 5.0x10(-6)g mL(-1) and 5.0x10(-7) to 1.0x10(-5)g mL(-1) for IAA and IBA, respectively. The detection limits were 2.0x10(-8)g mL(-1) and 2.0x10(-7)g mL(-1) for IAA and IBA, respectively. The relative standard deviation (RSD) of intra-day were 3.1% and 2.3% (n=11) for 2x10(-6)g mL(-1) IAA and 2x10(-6)g mL(-1) IBA; The relative standard deviations of inter-day precision were 6.9% and 4.9% for 2x10(-6)g mL( 1) IAA and 2x10(-6)g mL(-1) IBA. The proposed method had been successfully applied to the determination of auxin in mung bean sprouts. PMID- 19559869 TI - Biosensor for chlorogenic acid based on an ionic liquid containing iridium nanoparticles and polyphenol oxidase. AB - A biosensor based on the ionic liquid, 1-n-butyl-3-methylimidazolium hexafluorophosphate containing dispersed iridium nanoparticles (Ir-BMI.PF(6)) and polyphenol oxidase was constructed. This enzyme was obtained from the sugar apple (Annona squamosa), immobilized in chitosan ionically crosslinked with oxalate. The biosensor was used for determination of chlorogenic acid by square wave voltammetry. The polyphenol oxidase catalyzes the oxidation of chlorogenic acid to the corresponding o-quinone, which is electrochemically reduced back to this substance at +0.25V vs. Ag/AgCl. Under optimized operational conditions the chlorogenic acid concentration was linear in the range of 3.48x10(-6) to 4.95x10( 5)mol L(-1) with a detection limit of 9.15x10(-7)mol L(-1). The biosensor was applied in the determination of chlorogenic acid in organic and decaffeinated coffee and the results compared with those obtained using the capillary electrophoresis method. The recovery study for chlorogenic acid in these samples gave values of 93.2-105.7%. PMID- 19559870 TI - Determination of thiosulfate in human urine by high performance liquid chromatography. AB - Thiosulfate is a sulfate analogue with a thiosulfur substituent and is found in human samples. Its concentration in urine is increased in some diseases and after exposure to hydrogen sulfide gas. We have developed a sensitive, simple and cheap method for thiosulfate determination in urine. The method is based on precolumn derivatization with 2-chloro-1-methylquinolinium tetrafluoroborate followed by reversed-phase liquid chromatography separation and ultraviolet detection of 1 methyl-2-thioquinolone at 375 nm. The calibration curve for thiosulfate was linear in the tested range 0.5-50 micromol L(-1) with correlation coefficient better than 0.999. The analytical recovery and relative standard deviation values for precision within the calibration range were from 90.1% to 104.2% and from 2.39% to 5.59%, respectively. The lower limit of detection and quantitation were 0.3 and 0.5 micromol L(-1), respectively. The mean (range) concentration of thiosulfate normalized against creatinine for apparently healthy seven women and six men was 2.21 (1.45-2.77) and 2.51 (1.36-4.89)mmol mol(-1) creatinine, respectively. We monitored thiosulfate in urine samples from one volunteer for 24 h. The urinary excretion of thiosulfate was 21.4 micromol per 24 h. This method can be used for routine clinical monitoring thiosulfate in urine. Cysteine and cysteinylglycine can be measured concurrently, if needed. PMID- 19559871 TI - Trace iodine quantitation in biological samples by mass spectrometric methods: the optimum internal standard. AB - Accurate quantitation of iodine in biological samples is essential for studies of nutrition and medicine, as well as for epidemiological studies for monitoring intake of this essential nutrient. Despite the importance of accurate measurement, a standardized method for iodine analysis of biological samples is yet to be established. We have evaluated the effectiveness of (72)Ge, (115)In, and (129)I as internal standards for measurement of iodine in milk and urine samples by induction coupled plasma mass spectrometry (ICP-MS) and of (35)Cl(18)O(4)(-), (129)I(-), and 2-chlorobenzenesulfonate (2-CBS) as internal standards for ion chromatography-tandem mass spectrometry (IC-MS/MS). We found recovery of iodine to be markedly low when IC-MS/MS was used without an internal standard. Percent recovery was similarly low using (35)Cl(18)O(4) as an internal standard for milk and unpredictable when used for urine. 2 Chlorobenzebenzenesulfonate provided accurate recovery of iodine from milk, but overestimated iodine in urine samples by as much as a factor of 2. Percent recovery of iodine from milk and urine using ICP-MS without an internal standard was approximately 120%. Use of (115)In predicted approximately 60% of known values for both milk and urine samples. (72)Ge provided reasonable and consistent percent recovery for iodine in milk samples (approximately 108%) but resulted in approximately 80% recovery of iodine from urine. Use of (129)I as an internal standard resulted in excellent recovery of iodine from both milk and urine samples using either IC-MS/MS and ICP-MS. PMID- 19559872 TI - Electrochemical determination of arsenite in neutral media on reusable gold nanostructured films. AB - We report a simple and novel method of stirring-only-driven accumulation and electrochemical determination of arsenite (As(III)) with both of the oxidation and reduction peaks associated with As(0)/As(III) using a gold nanofilm electrode in neutral solution. Under stirring, a large amount of As(III) was deposited on the modified electrode and the electrochemical response was greatly amplified. The accumulated As(III) on the electrode showed well-defined redox couple in 0.1M blank phosphate buffer solution (pH 7.0), which could be used for the measurement of As(III). Under optimal conditions, As(III) could be detected in the range from 0.20 to 375 ppb with a detection limit of 0.04 ppb. In particular, with the use of the reduction peak of As(III) the modified electrode exhibits excellent performance for As(III) determination even in the presence of abundant Cu(II). The regeneration of the electrodes is facile with good reproducibility. The electrochemical system was applied to analyze As(III) in lake water, As(III) spiked tap water and drinking water. PMID- 19559873 TI - Solid phase extraction of Co ions using L-tyrosine immobilized on multiwall carbon nanotubes. AB - A study was performed to assess the performance of aminoacids immobilized on carbon nanotubes (CNTs) for their employment as a sorbent for solid phase extraction systems. An immobilization method is introduced and the aminoacid L tyrosine was chosen as a case study. A spectrophotometric study revealed the amount of aminoacid immobilizated on CNTs surface, and it turned to be of 3174 micromol of L-tyrg(-1). The material was tested for Co retention using a minicolumn inserted in a flow system. At pH 7.0, the amount of Co retained by the column was of 37.58+/-3.06 micromol Co g(-1) of CNTs. A 10% (v/v) HNO(3) solution was chosen as eluent. The pH study revealed that Co binding increased at elevated pH values. The calculation of the mol ratio (moles of Co bound at pH 9 to moles of l-tyr) turned to be 3:1. The retention capacity was compared to other bivalent cations and showed the following tendency: Cu(2+)>Ni(2+)>Zn(2+)>>Co(2+). The analytical performance was evaluated and an enrichment factor of 180 was obtained when 10 mL of 11.37 microg L(-1)Co solution was loaded onto the column at pH 9.0; reaching a limit of detection (LoD) of 50 ng L(-1). The proposed system was successfully applied to Co determination in QC-LL2 standard reference material (metals in natural water). PMID- 19559874 TI - Metabolomic profiling of rat serum associated with isoproterenol-induced myocardial infarction using ultra-performance liquid chromatography/time-of flight mass spectrometry and multivariate analysis. AB - An ultra-performance liquid chromatography/time-of-flight mass spectrometry (UPLC/TOF-MS)-based metabolomic approach was developed to characterize the metabolic profile associated with isoproterenol (ISO)-induced myocardial infarction (MI). Analysis of the serum samples revealed distinct changes in the biochemical patterns of ISO-induced rats. A multivariate statistical method, supervised partial least squares-discriminant analysis (PLS-DA), was then used for screening of potential biomarkers. As a result, 13 lipid biomarkers, including lysophosphatidylcholines (Lyso-PCs) and fatty acids were identified by the accurate mass measurement of TOF-MS. The relationship between abnormal lipid metabolism and the formation of MI were also studied. This work demonstrates the utility of UPLC/TOF-MS-based metabolic profiling combined with multivariate analysis as a powerful tool to further investigate the pathogenesis of cardiovascular diseases. PMID- 19559875 TI - Variable selection using probability density function similarity for support vector machine classification of high-dimensional microarray data. AB - One problem with discriminant analysis of microarray data is representation of each sample by a large number of genes that are possibly irrelevant, insignificant or redundant. Methods of variable selection are, therefore, of great significance in microarray data analysis. To circumvent the problem, a new gene mining approach is proposed based on the similarity between probability density functions on each gene for the class of interest with respect to the others. This method allows the ascertainment of significant genes that are informative for discriminating each individual class rather than maximizing the separability of all classes. Then one can select genes containing important information about the particular subtypes of diseases. Based on the mined significant genes for individual classes, a support vector machine with local kernel transform is constructed for the classification of different diseases. The combination of the gene mining approach with support vector machine is demonstrated for cancer classification using two public data sets. The results reveal that significant genes are identified for each cancer, and the classification model shows satisfactory performance in training and prediction for both data sets. PMID- 19559876 TI - Self-modeling curve resolution method applied for the evaluation of dissolution testing data: a case study of meloxicam-mannitol binary systems. AB - This paper introduces some chemometric methods, i.e., self-modeling curve resolution (SMCR), multivariate curve resolution-alternating least squares (MCR ALS) and parallel factor analysis (PARAFAC and PARAFAC2), which are used to evaluate in vitro dissolution testing data detected by a UV-vis spectrophotometer on meloxicam-mannitol binary systems. These systems were chosen because of their relative simplicity to apply as part of the validation process illustrating the effectiveness of the developed and applied chemometric method. The paper illustrates the failure of PARAFAC methods used before for pharmaceutical data evaluations as well, and we suggest application of the feasible band form given by SMCR as a more general procedure. Steps to improve the dissolution behavior of drugs have become among the most interesting aspects of pharmaceutical technology, and our results show that a larger particle size of meloxicam is advantageous for dissolution. Instead of the use of only one characteristic wavelength, appropriate chemometric methods can furnish more information from dissolution testing data, i.e., the individual dissolution rate profiles and the individual spectra for all the components can be obtained without resorting to any separation techniques such as HPLC. PMID- 19559877 TI - Enzyme class identification in cleaning products by hydrolysis followed by derivatization with o-phthaldialdehyde, HPLC and linear discriminant analysis. AB - The enzymes present in raw materials of the cleaning industry (enzyme industrial concentrates) and in household cleaners were isolated by precipitation with acetone and hydrolyzed with HCl. The resulting amino acids were derivatized with o-phthaldialdehyde, and the derivatives were separated by HPLC. The peaks of 14 amino acids were observed using a C18 column and a multi-segmented gradient of acetonitrile-water in the presence of a 5 mM citric/citrate buffer of pH 6.5. Using either normalized peak areas (divided by the sum of the peak areas of the chromatogram) or ratios of pairs of peak areas as predictor variables, linear discriminant analysis models, capable of predicting the enzyme class, including proteases, lipases, amylases and cellulases, were constructed. For this purpose, both enzyme industrial concentrates and detergent bases spiked with them were included in the training set. In all cases, the enzymes of the evaluation set, including industrial concentrates, spiked detergent bases and commercial cleaners were correctly classified with assignment probabilities higher than 99%. PMID- 19559878 TI - Development of an automatic solid phase extraction and liquid chromatography mass spectrometry method by using a monolithic column for the analysis of Cyclosporin A in human plasma. AB - A sensitive and specific and automated liquid chromatography-electrospray mass spectrometric (LC-ESI-MS) assay for the quantification of Cyclosporin A in human plasma was developed. Following a simple protein precipitation step, the supernatant was extracted on-line and directly injected into the system LC-ESI MS. A relatively new type of monolithic column consisting of a silica rod with bimodal pore structure was used to achieve a retention time of 2.4 min with a very low backpressure at a flow rate of 1 ml/min. The assay was linear from 0.050 to 1.000 microg/ml. The mean recovery was 91%. The mean inter-day and intra-day precisions were 1.85% and 2.83%, respectively. The combination of the automated solid phase extraction and the low retention time achieved with this columns increase the throughput and decrease the time of analysis of each sample. This technology is useful in order to improve the efficiency of the bioanalytical studies. PMID- 19559879 TI - Supercritical fluid extraction--achiral liquid chromatography with circular dichroism detection for the determination of menthone enantiomers in natural peppermint oil samples. AB - A simple and enantioselective method for the determination of menthone enantiomers in peppermint essential oil samples is proposed. The method involves the initial supercritical fluid extraction (SFE) to clean-up and extraction of analytes and their preconcentration on C(18) adsorption cartridges followed by achiral liquid chromatographic separation and direct circular dichroism (CD) detection. The calibration curve of the anisotropy factor (g) versus the enantiomeric excess was linear, with a correlation coefficient (R(2)) of 0.9970. The precision evaluated by UV peak area and CD peak area was suitable both in terms of intra- and inter-day precision (RSD<5.1% in all cases). The usefulness of the proposed method was demonstrated by analyzing natural and spiked peppermint oil samples. This method has the advantages of being rapid and precise without using an expensive chiral column. It was demonstrated to be suitable for the simultaneous determination of both enantiomers and for assessing the chemical purity of menthone. PMID- 19559880 TI - Electrochemical characterization of a superoxide biosensor based on the co immobilization of cytochrome c and XOD on SAM-modified gold electrodes and application to garlic samples. AB - This paper describes the characterization and optimization of an amperometric cytochrome c (cyt c)-based sensor for the determination of the antioxidant capacity of pure substances and natural samples. The cyt c and the xanthine oxidase (XOD) enzyme were co-immobilized on the electrode using the combination of several long-chain thiols. The self-assembled monolayer (SAM) was optimized in terms of composition and ratio between thiols. The immobilization protocol for both cyt c and XOD and the SAM formation time were evaluated through electrochemical methods, such as cyclic voltammetry (CV), square wave voltammetry (SWV), chronoamperometry (CA) and impedance spectroscopy (IS). Finally, the biosensor was applied to the determination of the antioxidant capacity of pure alliin and two compounds extracted from garlic bulbs. PMID- 19559881 TI - A self-assembled fusion protein-based surface plasmon resonance biosensor for rapid diagnosis of severe acute respiratory syndrome. AB - A surface plasmon resonance (SPR)-based biosensor was developed for simple diagnosis of severe acute respiratory syndrome (SARS) using a protein created by genetically fusing gold binding polypeptides (GBPs) to a SARS coronaviral surface antigen (SCVme). The GBP domain of the fusion protein serves as an anchoring component onto the gold surface, exploiting the gold binding affinity of the domain, whereas the SCVme domain is a recognition element for anti-SCVme antibody, the target analyte in this study. SPR analysis indicated the fusion protein simply and strongly self-immobilized onto the gold surface, through GBP, without surface chemical modification, offering a stable and specific sensing platform for anti-SCVme detection. AFM and SPR imaging analyses demonstrated that anti-SCVme specifically bound to the fusion protein immobilized onto the gold micropatterned chip, implying that appropriate orientation of bound fusion protein by GBP resulted in optimal exposure of the SCVme domain to the assay solution, resulting in efficient capture of anti-SCVme antibody. The best packing density of the fusion protein onto the SPR chip was achieved at the concentration of 10 microg mL(-1); this density showed the highest detection response (906RU) for anti-SCVme. The fusion protein-coated SPR chip at the best packing density had a lower limit of detection of 200 ng mL(-1) anti-SCVme within 10 min and also allowed selective detection of anti-SCVme with significantly low responses for non-specific mouse IgG at all tested concentrations. The fusion protein provides a simple and effective method for construction of SPR sensing platforms permitting sensitive and selective detection of anti-SCVme antibody. PMID- 19559882 TI - Electrochemical hydride generation of thallium. AB - An electrochemical hydride generation (ECHG) technique was developed to improve the determination of thallium by atomic spectrometry. The technique is based on the catholyte variation system for production of thallium hydride. Using Pb-Sn alloy as cathode, a transient peak shaped signal was achieved and its height, the maximum absorbance value, was taken as an analytical parameter. Parameters that might affect the hydride generation efficiency were investigated and the analytical performance of the method under the optimized experimental conditions was assessed. The linear range was 1-250 ng mL(-1) for thallium and the relative standard deviation of the method was 4.2% (RSD, n=7). The LOD for thallium was found to be 0.8 ng mL(-1), showing a significant improvement relative to conventional chemical hydride generation techniques. The proposed method was applied to the determination of thallium in unalloyed zinc standard reference material. This method offers high sensitivity, simplicity, rapidness, freeness from reagent and low acid consumption. PMID- 19559883 TI - Simultaneous determination of silicate and phosphate in environmental waters using pre-column derivatization ion-pair liquid chromatography. AB - A highly sensitive HPLC method for the simultaneous determination of soluble silicate and phosphate in environmental waters was developed, using ion-pair liquid chromatography preceded by the formation of their yellow alpha heteropolymolybdates. The moderate-pH mobile phase enabled to use a highly efficient reversed-phase silica column. The pre-column coloring reactions at moderate-pH were reproducible for both silicate and phosphate in all quantification ranges with R.S.D.s less than 2% and 5%, respectively. The linear calibration lines between concentrations (mg-SiO(2)/L and mg-PO(4)/L) and peak area intensities were obtained for silicate and phosphate both with acceptable determination coefficients (r(2)) of 0.9999. The limits of determination for both analytes were 0.007 mg-SiO(2)/L and 0.003 mg-PO(4)/L, which were calculated theoretically using 10sigma/slope. The four-digit dynamic ranges were obtained for 0.007-10mg-SiO(2)/L and 0.003-20mg-PO(4)/L. The developed method was applied for the analysis of tap water, river water, coastal seawater, well water, hot spring water, commercial mineral water, and laboratory water. The results were very reasonable and acceptable from the environmental viewpoints, which were well correlated with those confirmed by the molybdenum-blue spectrophotometry. PMID- 19559884 TI - Design and application of a novel integrated electrochemical hydride generation cell for the determination of arsenic in seaweeds by atomic fluorescence spectrometry. AB - An integrated electrochemical hydride generation cell, mainly composed of three components (a gas liquid separator, a graphite tube cathode and a reticulate Pt wire anode), was laboratory constructed and employed for the detection of arsenic by coupling to atomic fluorescence spectrometry. This integrated cell was free of ion-exchange membrane and individual anolyte, with the virtues of low-cost, easy assembly and environmental-friendly. Using flow injection mode, the sample throughput could come to 120 h(-1) attributed to the small dimension of the cathode chamber. The operating conditions for the electrochemical hydride generation of arsenic were investigated in detail and the potential interferences from oxygen or various ions were also evaluated. Under the optimized conditions, no obvious oxygen quenching effects were observed. The limit of detection of As (III) for the sample blank solution was 0.2 ng mL(-1) (3sigma) and the relative standard deviation was 3.1% for nine consecutive measurements of 5 ng mL(-1) As (III) standard solution. The calibration curve was linear up to 100 ng mL(-1). The accuracy of the method was verified by the determination of arsenic in the reference materials GBW08517 (Laminaria Japonica Aresch) and GBW10023 (Porphyra crispata) and the developed method was successfully applied to determine trace amounts of arsenic in edible seaweeds. PMID- 19559885 TI - Ni(II)-baicalein complex modified multi-wall carbon nanotube paste electrode toward electrocatalytic oxidation of hydrazine. AB - A modified electrode Ni(II)-BA-MWCNT-PE has been fabricated by electrodepositing nickel(II)-baicalein [Ni(II)-BA] complex on the surface of multi-wall carbon nanotube paste electrode (MWCNT-PE) in alkaline solution. The Ni(II)-BA-MWCNT-PE exhibits the characteristic of improved reversibility and enhanced current responses of the Ni(III)/Ni(II) couple compared with Ni(II)-BA-CPE. It also shows good electrocatalytic activity toward the oxidation of hydrazine. Kinetic parameters such as the electron transfer coefficient alpha, rate constant k(s) of the electrode reaction, the diffusion coefficient D of hydrazine and the catalytic rate constant k(cat) of the catalytic reaction are determined. Moreover, the catalytic currents present linear dependence on the concentration of hydrazine from 2.5 microM to 0.2 mM by amperometry. The detection limit and sensitivity are 0.8 microM and 69.9 microA mM(-1), respectively. The modified electrode for hydrazine determination is of the property of simple preparation, good stability, fast response and high sensitivity. PMID- 19559886 TI - Salicylaldehyde hydrazones as fluorescent probes for zinc ion in aqueous solution of physiological pH. AB - Salicylaldehyde hydrazones of 1 and 2 were synthesized and their potential as fluorescent probes for zinc ion was investigated in this paper. Both of the probes were found to show fluorescence change upon binding with Zn(2+) in aqueous solutions, with good selectivity to Zn(2+) over other metal ions such as alkali/alkali earth metal ions and heavy metal ions of Pb(2+), Cd(2+) and Hg(2+). They showed 1:2 metal-to-ligand ratio when their Zn(2+) complex was formed. By introducing pyrene as fluorophore, 2 showed interesting ratiometric response to Zn(2+). Under optimal condition, 2 exhibited a linear range of 0-5.0 microM and detection limit of 0.08 microM Zn(2+) in aqueous buffer, respectively. The detection of Zn(2+) in drinking water samples using 2 as fluorescent probe was successful. PMID- 19559887 TI - Characterization of PDMS-modified glass from cast-and-peel fabrication. AB - In glass/poly(dimethylsiloxane) (PDMS) hybrid microfluidic chips, two different fabrication approaches are used: photolithographic or solid ink molds, or cast and-peel methods. In the latter, a thin slab of PDMS is laid down and fluid channels are cut manually or by machine. The cast-and-peel approach has been used successfully for low-shear culture devices, among other applications. The main drawback, not reported to date, of cast-and-peel methods is that removal of PDMS (exposing the glass substrate) results in nanoscopic domains of PDMS still attached to the surface. This residual PDMS is not observable by eye, but affects the hydrophobicity of the device. Using contact angle measurement, atomic force and fluorescence microscopy, the changes in glass surfaces from the cast-and-peel technique were elucidated. This study demonstrates the enhanced protein (NeutrAvidin) adsorption on PDMS treated glass surfaces, and the potential influence of altered glass properties on microfluidic applications has been discussed as well. PMID- 19559888 TI - An approach by using near-infrared diffuse reflectance spectroscopy and resin adsorption for the determination of copper, cobalt and nickel ions in dilute solution. AB - Near-infrared spectroscopy (NIRS) has been proved to be a powerful analytical tool and used in various fields, it is seldom, however, used in the analysis of metal ions in solutions. A method for quantitative determination of metal ions in solution is developed by using resin adsorption and near-infrared diffuse reflectance spectroscopy (NIRDRS). The method makes use of the resin adsorption for gathering the analytes from a dilute solution, and then NIRDRS of the adsorbate is measured. Because both the information of the metal ions and their interaction with the functional group of resin can be reflected in the spectrum, quantitative determination is achieved by using multivariate calibration technique. Taking copper (Cu(2+)), cobalt (Co(2+)) and nickel (Ni(2+)) as the analyzing targets and D401 resin as the adsorbent, partial least squares (PLS) model is built from the NIRDRS of the adsorbates. The results show that the concentrations that can be quantitatively detected are as low as 1.00, 1.98 and 1.00 mg L(-1) for Cu(2+), Co(2+) and Ni(2+), respectively, and the coexistent ions do not influence the determination. PMID- 19559889 TI - Modified cupric reducing antioxidant capacity (CUPRAC) assay for measuring the antioxidant capacities of thiol-containing proteins in admixture with polyphenols. AB - Proteins are not considered as true antioxidants but are known to protect antioxidants from oxidation in various antioxidant activity assays. This study aims to investigate the contribution of proteins, especially thiol-containing proteins, to the observed overall antioxidant capacity measured by known methods. To determine the antioxidant properties of thiol-containing proteins, the CUPRAC method of antioxidant assay using the oxidizing reagent Cu(II)-neocuproine previously used for simultaneous analysis of cystine and cysteine was adopted. While the CUPRAC method is capable of determining all antioxidant compounds including thiols in complex sample matrices, the Ellman method of thiol quantitation basically does not respond to other antioxidants. The antioxidant quantities in the selected samples were assayed with the ABTS and FRAP methods as well as with the CUPRAC method. In all applied methods, the dilutions were made with a standard pH 8 buffer used in the Ellman method by substituting the Na(2)EDTA component of the buffer with sodium citrate. On the other hand, the standard CUPRAC protocol was modified by substituting the pH 7 ammonium acetate buffer (at 1M concentration) with 8M urea buffer adjusted to pH 7 by neutralizing with 6M HCl. Urea helps to partly solubilize and denaturate proteins so that their buried thiols be oxidized more easily. All methods used in the estimation of antioxidant properties of proteins (i.e., CUPRAC, Ellman, ABTS, and FRAP) were first standardized with a simple thiol compound, cysteine, by constructing the calibration curves. The molar absorptivities of these methods for cysteine were: epsilon(CUPRAC)=7.71x10(3), epsilon(Ellman)=1.37x10(4), epsilon(ABTS)=2.06x10(4), and epsilon(FRAP)=2.98x10(3)L mol(-1)cm(-1). Then these methods were applied to various samples containing thiols, such as glutathione (reduced form:GSH), egg white, whey proteins, and gelatin. Additionally, known quantities of selected antioxidants were added to these samples to show the additivity of responses. PMID- 19559890 TI - Strategies for the identification of urinary calculus by laser induced breakdown spectroscopy. AB - The present work studies two different strategies to identify urinary calculus. On one hand, (linear or parametric and rank or non-parametric) correlation methods using a mu-LIBS system are studied. On the other hand, elemental ratios of reference materials are determined by using a higher-energy laser and an Echelle spectrograph with an ICCD camera, although without microscope. A data treatment method was applied for each system and real samples of kidney stones- previously analyzed by IR spectroscopy--were used for reliable evaluation of two identification strategies. PMID- 19559891 TI - Electro-oxidation and determination of trazodone at multi-walled carbon nanotube modified glassy carbon electrode. AB - A simple and rapid electrochemical method was developed for the determination of trace-level trazodone, based on the excellent properties of multi-walled carbon nanotubes (MWCNTs). The MWCNT-modified glassy carbon electrode was constructed and the electrochemical behavior of trazodone was investigated in detail. The cyclic voltammetric results indicate that MWCNT-modified glassy carbon electrode can remarkably enhance electrocatalytic activity towards the oxidation of trazodone in neutral solutions. It leads to a considerable improvement of the anodic peak current for trazodone, and allows the development of a highly sensitive voltammetric sensor for the determination of trazodone. Trazodone could effectively accumulate at this electrode and produce two anodic peaks at about 0.73 V and 1.00 V. The electrocatalytic behavior was further exploited as a sensitive detection scheme for the trazodone determination by differential-pulse voltammetry. Under optimized conditions, the concentration range and detection limit are 0.2-10 microM and 24 nM, respectively for trazodone. The proposed method was successfully applied to trazodone determination in pharmaceutical samples. The analytical performance of this sensor has been evaluated for detection of analyte in urine as a real sample. PMID- 19559892 TI - Arsenic speciation in marine product samples: comparison of extraction-HPLC method and digestion-cryogenic trap method. AB - For the arsenic speciation in marine product samples, two types of pretreatment analysis combination were compared. One is the combination of solvent extraction and high performance liquid chromatography (HPLC) followed by a highly sensitive arsenic detection, while the other is the combination of alkaline digestion and cryogenic trap (CT) method followed by a highly sensitive arsenic detection. For six certified reference materials (CRMs) of marine animal samples, the concentrations of arsenobetaine (AsB) obtained from the extraction-HPLC method were very consistent with those of trimethylated arsenic species measured by the digestion-CT method. For four seaweed samples, the determination of three arsenosugars (Sugar-1, Sugar-2, and Sugar-3) was favorably carried out by the extraction-HPLC method. Those seaweed samples were also subjected to the digestion-CT method, and the amounts of dimethylated arsenic species measured by the method were approximately equal to the sum of the amounts of dimethylarsinic acid (DMAA) and three arsenosugars (Sugar-1+Sugar-2+Sugar-3) obtained from the extraction-HPLC method. PMID- 19559894 TI - Label-free aptamer-based chemiluminescence detection of adenosine. AB - We have developed a novel sensitive chemiluminescence (CL) aptasensor for the target assay as exemplified by using adenosine as a model target. In this work, we have demonstrated the signaling mechanism to make detection based on magnetic separation and 3,4,5-trimethoxyl-phenylglyoxal (TMPG), a special CL reagent as the signaling molecule, which reacts instantaneously with guanine nucleobases (G) of adenosine-binding aptamer strands. Briefly, amino-functioned capture DNA sequences are immobilized on the surface of carboxyl-modified magnetic beads, and then hybridized with label-free G-rich (including 15 guanine nucleobases) adenosine-binding aptamer strands to form our CL aptasensor. Upon the introduction of adenosine, the aptamer on the surface of magnetic beads is triggered to make structure switching to the formation of the adenosine/aptamer complex. Consequently, G-rich aptamer strands are forced to dissociate from magnetic beads sensing interface, resulting in a decrease of CL signal. The decrement of peak signal is proportional to the amount of adenosine. The effects of the amounts of capture DNA, aptamer, magnetic beads are investigated and optimized. It was found that the CL intensity had a linear dependency on the concentration of adenosine in the range of 4x10(-7) to 1x10(-5)M. With a low detection limit of 8x10(-8)M and simplicity in CL detection, this novel technique will offer a great promise for future target/aptamer analysis. PMID- 19559893 TI - Copolypeptide-doped polyaniline nanofibers for electrochemical detection of ultratrace trinitrotoluene. AB - This paper demonstrates a new electrochemical method for the detection of ultratrace amount of 2,4,6-trinitrotoluene (TNT) with synthetic copolypeptide doped polyaniline nanofibers. The copolypeptide, comprising of glutamic acid (Glu) and lysine (Lys) units, is in situ doped into polyaniline through the protonation of the imine nitrogen atoms of polyaniline by the free carboxylic groups of Glu segments, resulting in the formation of polyaniline nanofibers of emeraldine salt. The free amino groups of Lys segments at the surface of nanofibers provide the receptor sites of TNT through the formation of charge transfer complex between the electron-rich amino groups and the electron deficient aromatic rings. Adsorptive stripping voltammetry results demonstrate that the poly(Glu-Lys)-doped nanofibers confined onto glassy carbon electrodes exhibit a remarkable enriching effect and thus sensitive electrochemical response to TNT with a linear dynamic range of 0.5-10 microM and a detection limit down to 100 nM. Moreover, other kinds of nitro compounds show different redox behaviors from TNT at the doped nanofibers, and thus do not interfere with the electrochemical detection of TNT. This study essentially offers a new and simple method for electrochemical detection of ultratrace TNT. PMID- 19559895 TI - Correction of self-absorption effect in calibration-free laser-induced breakdown spectroscopy by an internal reference method. AB - A simplified procedure for correcting self-absorption effect was proposed in calibration-free laser-induced breakdown spectroscopy (CF-LIBS). In typical LIBS measurement conditions, the plasma produced is often optically thick, especially for the strong lines of major elements. The selection of self-absorption lines destroys the performance of CF-LIBS, and the familiar correction method based on the curve of growth is complex in implementation. The procedure we proposed, named internal reference for self-absorption correction (IRSAC), first chose an internal reference line for each species, then compared other spectral line intensity of the same species with the reference line to estimate the self absorption degrees of other spectral lines, and finally achieved an optimal correction by a regressive algorithm. The self-absorption effect of the selected reference line can be ignored, since the reference line with high excitation energy of the upper level is slightly affected by the self-absorption. Through the IRSAC method, the points on the Boltzmann plot become more regular, and the evaluations of the plasma temperature and material composition are more accurate than the basic CF-LIBS. PMID- 19559896 TI - Microwave-assisted digestion procedures for biological samples with diluted nitric acid: identification of reaction products. AB - Microwave-assisted sample preparation using diluted nitric acid solutions is an alternative procedure for digesting organic samples. The efficiency of this procedure depends on the chemical properties of the samples and in this work it was evaluated by the determination of crude protein amount, fat and original carbon. Soybeans grains, bovine blood, bovine muscle and bovine viscera were digested in a cavity-microwave oven using oxidant mixtures in different acid concentrations. The digestion efficiency was evaluated based on the determination of residual carbon content and element recoveries using inductively coupled plasma optical emission spectrometry (ICP OES). In order to determine the main residual organic compounds, the digests were characterized by nuclear magnetic resonance ((1)H NMR). Subsequently, studies concerning separation of nitrobenzoic acid isomers were performed by ion pair reversed phase liquid chromatography using a C18 stationary phase, water:acetonitrile:methanol (75:20:5, v/v/v)+0.05% (v/v) TFA as mobile phase and ultraviolet detection at 254 nm. Sample preparation based on diluted acids proved to be feasible and a recommendable alternative for organic sample digestion, reducing both the reagent volumes and the variability of the residues as a result of the process of decomposition. It was shown that biological matrices containing amino acids, proteins and lipids in their composition produced nitrobenzoic acid isomers and other organic compounds after cleavage of chemical bonds. PMID- 19559898 TI - Theoretical prediction of the native fluorescence of pharmaceuticals. AB - At present, to search fluorescent compounds or to increase the native fluorescence is an active research line specially and not only with analytical purposes. On some analytical areas and from the early times of applications of fluorescence (mid-fifties) the fluorimeter was defined as the suitable detector for determination of pharmaceuticals and subsequently, this detection mode has been widely applied. Therefore, it is mandatory to develop new strategies to discover or to enhance in a simple way the native fluorescence of organic compounds to increase the number of analytes to be determined by direct fluorescence. In the present paper are studied further applications of a new tool suitable to increase the research in analytical field. Calculations on molecular connectivity and discriminant analysis are applied to a certain number of pharmaceuticals (and some pesticides) on which fluorescence behaviour was observed in an experimental screening or obtained from scientific literature. The screening tests were based on the on-line fluorimetric measurement by using a continuous-flow assembly. The screening comprised pre-selected compounds with different molecular structures. The theoretical predictions agree with the empirical results from the screening test. PMID- 19559897 TI - Automated nucleic acids isolation using paramagnetic microparticles coupled with electrochemical detection. AB - Easy, efficient and low demanding separation of mRNA from biological material is needed to study gene expression and to use in chip technologies. It is common knowledge that each mRNA molecule contains sequence of 25 adenines. This feature can be used for binding mRNA on the surface of the particles coated by thymine chains. The present work reports on suggesting and optimizing of mRNA separation and detection from biological material via paramagnetic microparticles coupled with electrochemical detection. Primarily we optimized cyclic and square wave voltammetric conditions to detect poly(A), which was used as standard to mimic behaviour of mRNA. Under the optimized square wave voltammetric conditions (frequency 280 Hz, accumulation time 200 s, supporting electrolyte and its temperature: acetate buffer 4.6 and 35 degrees C) we estimated detection limit down to 1 ng of poly(A) per ml. To enhance effectiveness and repeatability of isolation of nucleic acid automated approach for rinsing and hybridizing was proposed. We optimized the whole procedure and experimental conditions. Using automated way of isolation and under optimized conditions the yield of poly(A) (isolated concentration of poly(A)/given concentration of poly(A)*100) was approximately 75%. The suggested and optimized method for poly(A) isolation and detection was utilized for the analysis of brain tissues of patients with traumatic brain injury. The total amount of isolated mRNA varied from 40 to 760 g of mRNA per g of brain tissue. The isolation of mRNA from six samples per run was not longer than 2.5h. Moreover, we applied the optimized procedure on fully automated pipetting instrument to isolate mRNA. The instrument was successfully tested on the analysis of extracts from roots of maize plants treated with cadmium(II) ions. PMID- 19559899 TI - Identification of reaction compounds in micrometric layers from gothic paintings using combined SR-XRD and SR-FTIR. AB - Synchrotron radiation X-ray diffraction (micro-SR-XRD) and Fourier transform infrared spectroscopy (micro-SR-FTIR) are used in the non-destructive identification of reaction and aging compounds from micrometric ancient painting layers. The combination of the micrometer size and non-destructive nature of the techniques together with the high resolution and brilliance of the synchrotron radiation has proved to be a procedure most advantageous for the study of reaction, aging and degradation processes. Copper, lead and calcium carboxylates and oxalates are determined in the chromatic, preparation and alteration layers from 15th century egg tempera and oil paintings. Their nature and crystallinity have been assessed. Some hypothesis about the mechanisms of development of both carboxylates and oxalates are presented. PMID- 19559900 TI - Preconcentration and determination of trace metals by flow injection micelle mediated extraction using flame atomic absorption spectrometry. AB - Micelle-mediated extraction/preconcentration is incorporated on-line into a flow injection system used to determine low levels of Cd(II), Co(II), Cu(II), Mn(II), Ni(II), Pb(II) and Zn(II) present in various samples. The analyte is complexed with HBDAP (N,N'-bis(2-hydroxy-5-bromo-benzyl)1,2-diaminopropane). Under optimal conditions, a solution of 30% (m/v) NaCl and a sample solution containing 2.5 mL of 1% (m/v) sodium dodecyl sulfate (SDS), 0.5 mL of 1.8x10(-3)M HBDAP and 2.5 mL of pH 8.5 borate buffer solution in 25 mL were pumped through the cotton filled mini-column; onto which the surfactant-rich phase containing the complex is collected. A solution of 0.5M HNO(3) in 50% acetone is used as the eluent. The limits of detection are (ng mL(-1)) Cd=0.39, Cu=3.2, Co=7.5, Mn=3.0, Ni=3.4, Pb=17.9 and Zn=0.89 if the sample is allowed to flow for 30s, but improved for extended preconcentration periods. Analysis of liquid and solid reference materials showed good agreement with the certified values. Complex formation constants between HBDAP and these metal ions were also determined potentiometrically. PMID- 19559901 TI - Determination of free captopril in human plasma by liquid chromatography with mass spectrometry detection. AB - A new simple, sensitive and selective liquid chromatography coupled with mass spectrometry (LC/MS) method for quantification of captopril after precolumn derivatization with p-bromo-phenacyl-bromide in human plasma was validated. Plasma samples were analysed on a monolithic column (Cromolith Performance-RP 18e, 100 mm x 4.6 mm I.D., 3 microm) under isocratic conditions using a mobile phase of a 40:60 (v/v) mixture of acetonitrile and 0.1% (v/v) formic acid in water. The flow rate was 1 mL/min at the column temperature of 30 degrees C. In these chromatographic conditions, the retention time was 4.4 min for captopril derivative. The detection of the analyte was in MRM mode using an ion trap mass spectrometer with electrospray positive ionisation. The monitored ions were 216, 253, 255, 268, 270 m/z derived from 415 m/z for derivatized captopril. The sample preparation was very simple and consisted in plasma protein precipitation from 0.2 mL plasma using 0.3 mL methanol after the derivatization reaction was completed. Calibration curves were generated over the range of 10-3000 ng/mL with values for coefficient of correlation greater than 0.993 and by using a weighted (1/y(2)) quadratic regression. The values for precision (CV %) and accuracy (relative error %) at quantification limit were less than 9.9% and 3.9%, for within- and between-run, respectively. The mean recovery of the analyte was 99%. Derivatized samples demonstrated good short-term, long-term, post-preparative and freeze-thaw stability. This is the first reported LC-MS/MS method for analysis of captopril in human plasma that uses protein precipitation as sample processing procedure. The method is very simple and allows obtaining a very good recovery of the analyte. The validated LC-MS/MS method has been applied to a pharmacokinetic study of 50mg captopril tablets on healthy volunteers. PMID- 19559902 TI - Determination of perchlorate in river by ion-pair hollow-fiber liquid-phase microextraction coupled with electrospray ionization tandem mass spectrometry. AB - This paper describes the analysis of perchlorate (ClO(4)(-)) in surface water samples by a rapid and reliable ion-pair hollow-fiber liquid-phase microextraction (HF-LPME) method coupled with flow-injection electrospray ionization tandem mass spectrometry (ESI-MS-MS) technique. The effects of the type and concentration of ion-pairing reagents, extraction time, temperature and pH value on the quantitative extraction of perchlorate by ion-pair HF-LPME were investigated and optimized. Di-n-hexyl ammonium acetate (DHAA) was employed to form an extractable ion-pair complex with aqueous perchlorate. The characteristic ions [ClO(4)-ClO(4)-DHA](-) at m/z 384.6 and 386.7 were observed in the ESI negative-ionization mode. The predominant product ions [ClO(4)](-) at m/z 99 and 101 were used for quantitation and to maximize the detection selectivity and sensitivity. The limit of detection (LOD) was 0.5 microg/L. The reliability and precision of the standard addition method of ion-pair HF-LPME for the determination of trace levels of perchlorate in surface water were demonstrated. PMID- 19559903 TI - Determination of the phenolic-group capacities of humic substances by non-aqueous titration technique. AB - The phenolic-group capacities of five humic substances, such as, the Aldrich humic acid, the humic and fulvic acids extracted from a soil, the humic and fulvic acids extracted from a peat have been precisely determined by the non aqueous potentiometric titration technique. The titration by KOH in the mixed solvent of DMSO:2-propanol:water=80:19.3:0.7 at [K(+)]=0.02 M enabled to measure the potential change in a wide range of pOH (=-log[OH(-)]), and thus to determine the capacities of phenolic groups which could not be precisely determined in the aqueous titration. The results of the titration revealed that the mean protonation constants of the phenolic groups were nearly the same for all humic substances and close to that of phenol in the same medium, indicating that each phenolic-group in the humic substances is rather isolated and is not electronically affected by other affecting groups in the humic macromolecule. PMID- 19559904 TI - Solid-state electrochemiluminescence sensor based on the Nafion/poly(sodium 4 styrene sulfonate) composite film. AB - An effective electrochemiluminescence (ECL) sensor based on Nafion/poly(sodium 4 styrene sulfonate) (PSS) composite film-modified ITO electrode was developed. The Nafion/PSS/Ru composite film was characterized by atomic force microscopy, UV-vis absorbance spectroscopy and electrochemical experiments. The Nafion/PSS composite film could effectively immobilize tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)(3)(2+)) via ion-exchange and electrostatic interaction. The ECL behavior of Ru(bpy)(3)(2+) immobilized in Nafion/PSS composite film was investigated using tripropylamine (TPA) as an analyte. The detection limit (S/N=3) for TPA at the Nafion/PSS/Ru composite-modified electrode was estimated to be 3.0 nM, which is 3 orders of magnitude lower than that obtained at the Nafion/Ru modified electrode. The Nafion/PSS/Ru composite film-modified indium tin oxide (ITO) electrode also exhibited good ECL stability. In addition, this kind of immobilization approach was simple, effective, and timesaving. PMID- 19559905 TI - Electrophoretic separation with 2-mm inner diameter fused-silica microcolumn packed with quartz microncrystals and its application. AB - The feasibility of a microcolumn electrophoresis technique was investigated with a 100mm length, 2mm I.D. fused-silica microcolumn packed with uniform quartz microncrystals prepared by hydrothermal synthesis. To evaluate the separation technique, tryptophan, phenylalanine and tyrosine were primarily separated by the microcolumn electrophoresis and detected at 216 nm without derivatization by an ordinary spectrophotometer. The separation conditions of the amino acids were optimized. With 1.5 mmol/L disodium phosphate buffer solution (pH 11.5) containing 25% (v/v) methanol and 10% (v/v) acetonitrile, the three amino acids were separated and the separation efficiency of tryptophan was 4.5x10(4)plates/m. The limits of detection were 0.035, 0.22 and 0.20 micromol/L, respectively. The sample capacity of the electrophoretic microcolumn achieved 35 microL. The proposed method was used to determine these amino acids in compound amino acid injection samples without derivatization. For the simplicity and portability of the microcolumn electrophoresis, it is studied as one of the high-performance separation techniques for an in situ and real-time electrokinetic flow analysis system. For its high detection sensitivity and large sample capacity, it can be developed for preparative electrophoresis. PMID- 19559906 TI - Analysis of aqueous pyrethroid residuals by one-step microwave-assisted headspace solid-phase microextraction and gas chromatography with electron capture detection. AB - A one-step microwave-assisted headspace solid-phase microextraction (MA-HS-SPME) has been applied to be a pretreatment step in the analysis of aqueous pyrethroid residuals by gas chromatography (GC) with electron capture detection (ECD). Microwave heating was applied to accelerate the vaporization of pyrethroids (bioallenthrin, bifenthrin, fenpropathrin, cyhalothrin, permethrin, cyfluthrin, cypermethrin, fluvalinate, fenvalerate and deltamethrin) into the headspace, and then being absorbed directly on a SPME fiber under the controlled conditions. Optimal conditions for the SPME sampling, such as the selection of sampling fiber, sample pH, sampling temperature and time, microwave irradiation power, desorption temperature and time were investigated and then applied to real sample analysis. Experimental results indicated that the extraction of pyrethroids from a 20-mL aquatic sample (pH 4.0) was achieved with the best efficiency through the use of a 100-microm PDMS fiber, microwave irradiation of 157 W and sampling at 30 degrees C for 10 min. Under optimum conditions, the detections were linear in the range of 0.05-0.5 microg/L with the square of correlation coefficients (R(2)) of >0.9913 for pyrethroids except bifenthrin being 0.9812. Method detection limits (MDL) were found to be varied from 0.2 to 2.6 ng/L for different pyrethroids based on S/N (signal to noise)=3. The coefficients of variation (CVs) for repeatability were 7-21%. A field underground water sample was analyzed with recovery between 88.5% to 115.5%. This method was proven to be a very simple, rapid, and solvent-free process to achieve the sample pretreatment before the analysis of trace pyrethroids in aqueous samples by gas chromatography. PMID- 19559907 TI - A new high-speed hollow fiber based liquid phase microextraction method using volatile organic solvent for determination of aromatic amines in environmental water samples prior to high-performance liquid chromatography. AB - A new and fast hollow fiber based liquid phase microextraction (HF-LPME) method using volatile organic solvents coupled with high-performance liquid chromatography (HPLC) was developed for determination of aromatic amines in the environmental water samples. Analytes including 3-nitroaniline, 3-chloroaniline and 4-bromoaniline were extracted from 6 mL basic aqueous sample solution (donor phase, NaOH 1 mol L(-1)) into the thin film of organic solvent that surrounded and impregnated the pores of the polypropylene hollow fiber wall (toluene, 20 microL), then back-extracted into the 6 mL acidified aqueous solution (acceptor phase, HCl 0.5 mol L(-1)) in the lumen of the two-end sealed hollow fiber. After the extraction, 5 microL of the acceptor phase was withdrawn into the syringe and injected directly into the HPLC system for the analysis. The parameters influencing the extraction efficiency including the kind of organic solvent and its volume, composition of donor and acceptor phases and the volume ratio between them, extraction time, stirring rate, salt addition and the effect of the analyte complexation with 18-crown-6 ether were investigated and optimized. Under the optimal conditions (donor phase: 6 mL of 1 mol L(-1) NaOH with 10% NaCl; organic phase: 20 microL of toluene; acceptor phase: 6 microL of 0.5 mol L(-1) HCl and 600 mmol L(-1) 18-crown-6 ether; pre-extraction and back-extraction times: 75 s and 10 min, respectively; stirring rate: 800 rpm), the obtained EFs were between 259 and 674, dynamic linear ranges were 0.1-1000 microg L(-1) (R>0.9991), and also the limits of detection were in the range of 0.01-0.1 micro gL(-1). The proposed procedure worked very well for real environmental water samples with microgram per liter level of the analytes, and good relative recoveries (91-102%) were obtained for the spiked sample solutions. PMID- 19559908 TI - Rapid analysis of captopril in human plasma and pharmaceutical preparations by headspace solid phase microextraction based on polypyrrole film coupled to ion mobility spectrometry. AB - A rapid, simple, and sensitive headspace solid phase microextraction coupled to ion mobility spectrometry (HS-SPME-IMS) method is presented for analysis of the highly specific angiotensin-converting enzyme (ACE) inhibitor, captopril (CAP). Positive ion mobility spectra of CAP were acquired with an ion mobility spectrometer equipped with a corona discharge ionization source. Mass-to-mobility correlation equation was used to identify product ions. A dodecylsulfate-doped polypyrrole (PPy-DS) coating was used as a fiber for SPME. The results showed that PPy-DS based SPME fiber was suitable for successfully extracting CAP from human blood plasma and pharmaceutical samples. The HS-SPME-IMS method provided good repeatability (R.S.D.s<4%) for aqueous and spiked plasma samples. The calibration graphs were linear in the range of 10-300 ng mL(-1) (R(2)>0.99) and detection limits were 7.5 ng mL(-1) for aqueous and 6.3 ng mL(-1) for plasma blank samples. Finally, a standard addition calibration method was applied to HS SPME-IMS technique for the analysis of blood plasma samples and tablets. Purpose method seemed to be suitable for the analysis of CAP in plasma samples as it is not time consuming (state total time from sample preparation to analysis), it required only small quantities of the sample, and no derivatization was required. PMID- 19559909 TI - Polyaniline and poly(flavin adenine dinucleotide) doped multi-walled carbon nanotubes for p-acetamidophenol sensor. AB - A conductive biocomposite film (MWCNTs-PANIFAD) which contains multi-walled carbon nanotubes (MWCNTs) along with the incorporation of poly(aniline) and poly(flavin adenine dinucleotide) co-polymer (PANIFAD) has been synthesized on gold and screen printed carbon electrodes by potentiostatic methods. The presence of MWCNTs in the MWCNTs-PANIFAD biocomposite film enhances the surface coverage concentration (Gamma) of PANIFAD and increases the electron transfer rate constant (k(s)) to 89%. Electrochemical quartz crystal microbalance studies reveal the enhancements in the functional properties of MWCNTs and PANIFAD present in MWCNTs-PANIFAD biocomposite film. Surface morphology of the biocomposite film has been studied using scanning electron microscopy and atomic force microscopy. The surface morphology results reveal that PANIFAD incorporated on MWCNTs. The MWCNTs-PANIFAD biocomposite film exhibits promising enhanced electrocatalytic activity towards the oxidation of p-acetamidophenol. The cyclic voltammetry has been used for the measurement of electroanalytical properties of p-acetamidophenol by means of PANIFAD, MWCNTs and MWCNTs-PANIFAD biocomposite film modified gold electrodes. The sensitivity value of MWCNTs-PANIFAD film (88.5 mA mM(-1)cm(-2)) is higher than the values which are obtained for PANIFAD (28.7 mA mM(-1)cm(-2)) and MWCNTs films (60.7 mA mM(-1)cm(-2)). Finally, the flow injection analysis (FIA) has been used for the amperometric detection of p acetamidophenol at MWCNTs-PANIFAD film modified screen printed carbon electrode. The sensitivity value of MWCNTs-PANIFAD film (3.3 mA mM(-1)cm(-2)) in FIA is also higher than the value obtained for MWCNTs film (1.1 mA mM(-1)cm(-2)). PMID- 19559910 TI - Determination of cadmium and lead species and phytochelatins in pea (Pisum sativum) by HPLC-ICP-MS and HPLC-ESI-MSn. AB - An analytical approach based on hyphenated techniques was used for studying the speciation of cadmium and lead in Pisum sativum. Proper preservation conditions were employed to avoid the oxidation of -SH groups and corresponding decomposition of metal-binding complexes. SEC column was washed with 5 mM beta mercaptoethanol and then samples were analysed using ICP-MS as a detector. Results showed that cadmium is the inhibitor of lead uptake. HPLC-ESI-MS(n) assays revealed fragmentation pathways of phytochelatins. PMID- 19559911 TI - Analysis of parabens in cosmetics by low pressure liquid chromatography with monolithic column and chemiluminescent detection. AB - This paper presents an application of chromatographic separation based on an ultra-short monolithic column and chemiluminescent detection in an FIA type instrument manifold for the determination of four paraben mixtures: methylparaben (MP), ethylparaben (EP), propylparaben (PP) and butylparaben (BP). The separation is achieved in 150 s using two consecutive carriers: first 12% ACN:water that changes 75 s after injection to 27% ACN:water. The detection is based on the oxidation of the hydrolysis product of parabens, p-hydroxybenzoic acid, with Ce(IV) in the presence of Rhodamine 6G which evokes chemiluminescence of sufficient intensity to enable a sensitive determination of these species. After optimization of the variables involved, the analytical method is characterized, displaying the following values for concentration ranges, detection limits and precision, as relative standard deviation at low concentration (0.15 mg l(-1)) MP: from 9.9x10(-7) to 3.3x10(-4)M; 1.9x10(-8); 5.6%; EP: from 9.0x10(-7) to 3.3x10(-4)M; 2.8x10(-8); 3.5%; PP: from 8.3x10(-7) to 9.9x10(-5)M; 2.3x10(-8); 4.2%; and BP: from 7.7x10(-7) to 9.9x10(-5)M; 4.2x10(-8)M; 6.2%. The method was applied and validated satisfactorily for the determination of these parabens in cosmetic samples, comparing the results against a liquid chromatography reference method. PMID- 19559912 TI - The use of Artificial Neural Networks for the selective detection of two organophosphate insecticides: chlorpyrifos and chlorfenvinfos. AB - Amperometric acetylcholinesterase (AChE) biosensors have been developed to resolve mixtures of chlorpyrifos oxon (CPO) and chlorfenvinfos (CFV) pesticides. Three different biosensors were built using the wild type from electric eel (EE), the genetically modified Drosophila melanogaster AChE B394 and B394 co immobilized with a phosphotriesterase (PTE). Artificial Neural Networks (ANNs) were used to model the combined response of the two pesticides. Specifically two different ANNs were constructed. The first one was used to model the combined response of B394+PTE and EE biosensors and was applied when the concentration of CPO was high and the other, modelling the combined response of B394+PTE and B394 biosensors, was applied with low concentrations of CPO. In both cases, good prediction ability was obtained with correlation coefficients better than 0.986 when the obtained values were compared with those expected for a set of six external test samples not used for training. PMID- 19559913 TI - Inductively coupled plasma atomic emission spectrometric determination of 27 trace elements in table salts after coprecipitation with indium phosphate. AB - The coprecipitation method using indium phosphate as a new coprecipitant has been developed for the separation of trace elements in table salts prior to their determination using inductively coupled plasma atomic emission spectrometry (ICP AES). Indium phosphate could quantitatively coprecipitate 27 trace elements, namely, Be, Ti, Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd, Pb, Sc, Y, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, and Lu, in a table salt solution at pH 10. The rapid coprecipitation technique, in which complete recovery of the precipitate was not required in the precipitate-separation process, was completely applicable, and, therefore, the operation for the coprecipitation was quite simple. The coprecipitated elements could be determined accurately and precisely by ICP-AES using indium as an internal standard element after dissolution of the precipitate with 5 mL of 1 mol L(-1) nitric acid. The detection limits (three times the standard deviation of the blank values, n=10) ranged from 0.001 microg (Lu) to 0.11 microg (Zn) in 300 mL of a 10% (w/v) table salt solution. The method proposed here could be applied to the analyses of commercially available table salts. PMID- 19559914 TI - Automated stopped-in-dual-loop flow analysis system for catalytic determination of vanadium in drinking water. AB - An automated stopped-in-dual-loop flow analysis (SIDL-FA) system is proposed for the determination of vanadium in drinking water. The chemistry is based on the vanadium-catalyzed oxidation reaction of p-anisidine by bromate in the presence of Tiron as an activator to produce a dye (lambda(max)=510 nm). A SIDL-FA system basically consists of a selection valve, three pumps (one is for delivering of standard/sample, and others are for reagents), two six-way injection valves, a spectrophotometric detector and a data acquisition device. A 100-microL coiled loop around a heated device is fitted onto each six-way injection valve. A well mixed solution containing reagents and standard/sample is loaded into the first loop on a six-way valve, and then the same solution is loaded into the second loop on another six-way valve. The solutions are isolated by switching these two six-way valves, so that the catalytic reaction can be promoted. The net waste can be zero in this stage, because all pumps are turned off. Then each resulting solution is dispensed to the detector with suitable time lag. A touchscreen controller is developed to automatically carry out the original SIDL-FA protocol. The proposed SIDL-FA method allows vanadium to be quantified in the range of 0.1 2 microg L(-1) and is applied to the determination of vanadium in drinking water samples. PMID- 19559915 TI - Rapid IC-ICP/MS method for simultaneous analysis of iodoacetic acids, bromoacetic acids, bromate, and other related halogenated compounds in water. AB - Haloacetic acids (HAAs) and bromate are toxic water disinfection by-products (DBPs) that the U.S. Environmental Protection Agency has regulated in drinking water. Iodoacetic acids (IAAs) are the emerging DBPs that have been recently found in disinfected drinking waters with higher toxicity than their corresponding chloro- and bromo-acetic acids. This study has developed a new rapid and sensitive method for simultaneous analysis of six brominated and four iodinated acetic acids, bromate, iodate, bromide, and iodide using ion chromatography-inductively coupled plasma-mass spectrometry (IC-ICP-MS). Mono-, di- and tri-chloroacetic acids are not detected by this method because the sensitivity of ICP-MS analysis for chlorine is poor. Following IC separation, an Elan DRC-e ICP-MS was used for detection, with quantitation utilizing m/z of 79, 127, and 74 amu for Br, I, and Ge (optional internal standard) species, respectively. Although the primary method used was an external standard procedure, an internal standard method approach is discussed herein as well. Calibration and validation were done in a variety of natural and disinfection treated water samples. The method detection limits (MDLs) in natural water ranged from 0.33 to 0.72 microg L(-1) for iodine species, and from 1.36 to 3.28 microg L(-1) for bromine species. Spiked recoveries were between 67% and 123%, while relative standard deviations ranged from 0.2% to 12.8% for replicate samples. This method was applied to detect the bromine and iodine species in drinking water, groundwater, surface water, and swimming pool water. PMID- 19559916 TI - Comparative evaluation of Dy(III) selective poly(vinyl) chloride based membrane electrodes of macrocyclic tetraimine Schiff's bases. AB - Three different derivatives of macrocyclic tetraimine Schiff's base have been synthesized and explored as a neutral ionophores for preparing poly(vinyl chloride) based membrane sensors selective to Dy(3+). The addition of sodium tetraphenyl borate and various plasticizers, viz., o-NPOE, DBP, DBBP, DOP and CN has been found to substantially improve the performance of the sensors. The best performance was obtained with the sensor no. 1 having membrane of Schiff's base (SL-1) with composition (w/w) SL-1 (4.5%): PVC (30.5%): o-NPOE (59.5%): NaTPB (5.5%). This sensor exhibits Nernstian response with slope 19.4 mV/decade of activity in the concentration range of 10(-8) to 1.0x10(-2)M Dy(3+), performs satisfactorily over wide pH range of (2.8-7.2) with a fast response time (10s). The sensor was also found to work satisfactorily in partially non-aqueous media up to 20% (v/v) content of acetonitrile, methanol or ethanol. The proposed sensor can be used over a period of 1.5 months without significant drift in potentials. The sensor has been also utilized for the determination of Dy(3+) level in different soil samples. PMID- 19559917 TI - Simultaneous chemiluminescence determination of promazine and fluphenazine using support vector regression. AB - In this work, chemiluminescence (CL) behaviors of two selected phenothiazines, namely promazine and fluphenazine hydrochloride, were investigated for their simultaneous determination using oxidation of Ru(bipy)(3)(2+) by Ce(4+) ions in acidic media. This method is based on the kinetic distinction of the CL reactions of fluphenazine and promazine with Ru(bipy)(3)(2+) and Ce(4+) system in a sulfuric acid medium. Least square support vector regression models were constructed for relating concentrations of both compounds to their CL profiles. The parameters of the model consisting of sigma(2) and gamma were optimized using all possible combinations of sigma(2) and gamma to select the model with the minimum root mean square cross validation. Under optimized conditions, the univariate calibration curve was linear over the concentration ranges of 0.4-30.0 microg mL(-1) and 0.07-5.0 microg mL(-1) with detection limits of 0.1 microg mL( 1) and 0.04 microg mL(-1) for promazine and fluphenazine, respectively. The influence of potential interfering substances on the determination of promazine and fluphenazine were studied. The proposed method was used for simultaneous determination of both compounds in synthetic mixtures and in spiked human plasma. PMID- 19559918 TI - Determination of senkirkine and senecionine in Tussilago farfara using microwave assisted extraction and pressurized hot water extraction with liquid chromatography tandem mass spectrometry. AB - Tussilago farfara (Kuan Donghua) is an important Chinese herbal medicine which has been shown to contain many bioactive compounds and widely used to relieve cough and resolve phlegm. However, besides therapeutic bioactive compounds, this herb has been found to contain toxic pyrrolizidine alkaloids (PAs), mainly senkirkine and traces of senecionine. In this report, conditions for microwave assisted extraction (MAE) and pressurized hot water extraction (PHWE) were optimized for the extraction of the PAs. The results were compared against heating under reflux. It was found that the binary mixture of MeOH:H(2)O (1:1) acidified using HCl to pH 2-3 was the optimal solvent for the extraction of the PAs in the plant materials. Liquid chromatography (LC) with ultra-violet (UV) detection and electrospray ionization mass spectrometry (ESI-MS) in the positive mode was used for the determination and quantitation of senkirkine and senecionine in the botanical extract. The proposed extraction methods with LC/MS allow for the rapid detection of the major and the minor alkaloids in T. farfara in the presence of co-eluting peaks. With LC/MS, the quantitative analysis of PAs in the extract was done using internal standard calibration and the precision was found to vary from 0.6% to 5.4% on different days. The limits of detection (LODs) and limits of quantitation (LOQs) for MAE and PHWE were found to vary from 0.26 microg/g to 1.04 micro/g and 1.32 micro/g to 5.29 microg/g, respectively. The method precision of MAE and PHWE were found to vary from 3.7% to 10.4% on different days. The results showed that major and minor alkaloids extracted using MAE and PHWE were comparable to that by heating under reflux. Our data also showed that significant ion suppression was not observed in the analysis of senkirkine and senecionine in the botanical extracts with co-eluting peaks. PMID- 19559919 TI - Colorimetric and fluorescent sensing of biologically important fluoride in physiological pH condition based on a positive homotropic allosteric system. AB - A novel positive homotropic allosteric system 1 based on 3-methylpyrozole-5-one-4 one-2',4'-dinitrophenylhydrazone was designed, synthesized and characterized. Colorimetric and fluorescent sensing of anions was achieved in physiological condition (pH 7.4), resulting from the positive homotropic allosterism of 1 induced by anions tested. In particular, the compound 1 exhibited a two-step response to the strong basic anions such as F(-). In the first step, the hydrazone form of 1 interacted with anions through hydrogen bonding with an obvious color change from yellow to orange upon addition of 0.3 equiv. of anions. In the second step, with further addition of anions, the hydrazone form of 1 was shifted to the azophenol form, whose anion binding was accompanied with an orange to-purple color change. In addition, the receptor 1 exhibited a fluorescent enhancement response to anions exploiting two possible signaling transduction mechanisms: (1) inhibition of photoinduced electronic transfer (PET) and (2) binding-induced rigidity of the host molecule. PMID- 19559920 TI - Comments on "Determination of S-nitrosoglutathione and other nitrosothiols by p hydroxymercurybenzoate derivatization and reverse phase chromatography coupled with chemical vapor generation atomic fluorescence detection" by Bramanti et al. PMID- 19559922 TI - In this issue... PMID- 19559923 TI - Research news and notes. PMID- 19559926 TI - Clinical experience with Matrix2 360 degrees coils in the treatment of 100 intracranial aneurysms. AB - BACKGROUND: The M2-360 degrees is a recent class of aneurysm coil. This device combines the second generation of bioactive copolymer coating, which is intended to promote aneurysm fibrosis, with the "360 degrees " design, which is meant to improve uniformity and density of packing. This study evaluates the safety and angiographic stability of these devices. METHODS: This was a retrospective review of 86 consecutive patients with 100 intracranial aneurysms that were treated using M2-360 degrees s. Follow-up was done at 6 and 12 months. RESULTS: Seventy eight aneurysms were coiled solely with M2-360 degrees s, and 22 aneurysms were treated with a combination of coils. In mixed-coil cases, the average percentage of coil volume consisting of M2-360 degrees coils was 78%. Procedure-related neurologic complications occurred in 6 patients (7%). Initial complete occlusion was obtained in 80 aneurysms. Of 76 aneurysms with 6-month angiographic follow up, 4 (5.3%) revealed further occlusion, 54 (71.1%) were unchanged, and 18 (23.7%) showed recanalization. Of 38 aneurysms with 12-month follow-up, 1 (2.6%) revealed further occlusion, 23 (60.5%) were unchanged, and 14 (36.8%) showed recanalization. Six- and 12-month angiograms showed major recanalization (requiring further coiling) in 3.9% and 15.8% of cases, respectively. CONCLUSIONS: The risk of complications with M2-360 degrees -treated aneurysms is comparable with reports of other coils, indicating that M2-360 degrees s are relatively safe. Although the initial occlusion rate is higher than that in other coiling series, recanalization rates were similar to those obtained with other coil designs. This study does not demonstrate an advantage with M2-360 degrees s. PMID- 19559927 TI - The role of collagen type I alpha2 polymorphisms: intracranial aneurysms in Koreans. AB - BACKGROUND: The COL1A2 is located on chromosome 7q22.1, and mutations in this gene have been associated with the development of IAs. In this study, we investigated whether the rs42524 and rs2621215 polymorphisms of the COL1A2 gene are associated with the development of cerebral aneurysms in the Korean population. METHODS: This was a hospital-based case control study conducted at Chonnam University Hospital, Gwangju, Korea. The study population consisted of 320 patients who had been treated for IA and 189 healthy hospital-based controls (angiographically negative for an IA). Two polymorphic loci were amplified by polymerase chain reaction, namely, rs42524 in exon 28 and rs2621215 in intron 46 of the COL1A2 gene, and analyzed by RFLP using HhaI or BfaI restriction enzymes, respectively. RESULTS: The genotype frequencies of rs42524 in cases were 88.0%, 11.4%, and 0.6% for the GG, GC, and CC genotypes, respectively, and in controls were 88.9%, 10.0%, and 1.1%, respectively. Similarly, the genotype frequencies of rs2621515 in cases were 88.0%, 10.1%, and 0.2% for the TT, TG, and GG genotypes, respectively, and among controls were 92.1%, 7.9%, and 0%, respectively. CONCLUSION: The rs2621215 SNP in intron 46 of the COL1A2 gene was found to be marginally associated with an increased risk of IA development in the Korean population examined. In contrast, rs42524 showed no association with an increased risk of IA development. PMID- 19559924 TI - Association of Chiari malformation type I and tethered cord syndrome: preliminary results of sectioning filum terminale. AB - OBJECTIVE: The pathogenesis of CM-I is incompletely understood. We describe an association of CM-I and TCS that occurs in a subset of patients with normal size of the PCF. METHODS: The prevalence of TCS was determined in a consecutively accrued cohort of 2987 patients with CM-I and 289 patients with low-lying cerebellar tonsils (LLCT). Findings in 74 children and 244 adults undergoing SFT were reviewed retrospectively. Posterior cranial fossa size and volume were measured using reconstructed 2D computed tomographic scans and MR images. Results were compared to those in 155 age- and sex-matched healthy control individuals and 280 patients with generic CM-I. The relationships of neural and osseus structures at the CCJ and TLJ were investigated morphometrically on MR images. Intraoperative CDU was used to measure anatomical structures and CSF flow in the lumbar theca. RESULTS: Tethered cord syndrome was present in 408 patients with CM I (14%) and 182 patients with LLCT (63%). In 318 patients undergoing SFT, there were no significant differences in the size or volume of the PCF as compared to healthy control individuals. Morphometric measurements demonstrated elongation of the brain stem (mean, 8.3 mm; P < .001), downward displacement of the medulla (mean, 4.6 mm; P < .001), and normal position of the CMD except in very young patients. Compared to patients with generic CM-I, the FM was significantly enlarged (P < .001). The FT was typically thin and taut (mean transverse diameter, 0.8 mm). After SFT, the cut ends of the FT distracted widely (mean, 41.7 mm) and CSF flow in the lumbar theca increased from a mean of 0.7 cm/s to a mean of 3.7 cm/s (P < .001). Symptoms were improved or resolved in 69 children (93%) and 203 adults (83%) and unchanged in 5 children (7%) and 39 adults (16%) and, worse, in 2 adults (1%) over a follow-up period of 6 to 27 months (mean, 16.1 months +/- 4.6 SD). Magnetic resonance imaging 1 to 18 months after surgery (mean, 5.7 months +/- 3.8 SD) revealed upward migration of the CMD (mean, 5.1 mm, P < .001), ascent of the cerebellar tonsils (mean, 3.8 mm, P < .001), reduction of brain stem length (mean, 3.9 mm, P < .001), and improvement of scoliosis or syringomyelia in some cases. CONCLUSIONS: Chiari malformation type I/TCS appears to be a unique clinical entity that occurs as a continuum with LLCT/TCS and is distinguished from generic CM-I by enlargement of the FM and the absence of a small PCF. Distinctive features include elongation and downward displacement of the hindbrain, normal position of the CMD, tight FT, and reduced CSF flow in the lumbar theca. There is preliminary evidence that SFT can reverse moderate degrees of tonsillar ectopia and is appropriate treatment for cerebellar ptosis after Chiari surgery in this cohort. PMID- 19559929 TI - Platelet-derived growth factor expression correlates with tumor grade and proliferative activity in human oligodendrogliomas. AB - BACKGROUND: For the last one and a half decade, it has been found that platelet derived growth factor (PDGF) promotes glial tumor growth through autocrine and paracrine loops, by expression of PDGFalpha receptor (PDGFRalpha) on glioma cells and PDGFbeta receptor (PDGFRbeta) on proliferating endothelial cells. However, studies on oligodendrogliomas, correlating expression of PDGF and its receptor with tumor grade and proliferative activity, through MIB-1 labeling index (LI) are relatively few as compared to astroglial counterpart. METHODS: Formalin-fixed paraffin-embedded tissues from 55 cases of oligodendrogliomas (34 World Health Organization [WHO] grade II and 21 WHO grade III tumors) were subjected to immunohistochemistry. MIB-1 LI was calculated, and a semiquantitative scoring system for expression of PDGF and PDGFRalpha was used. RESULTS: MIB-1 LI and PDGF expression increased with histologic grades of malignancy ("t" test, P < .001 and Mann Whitney test, U = 109, P < .001 respectively). The PDGF expression scores had a positive correlation with MIB-1 LI, irrespective of tumor grade (Pearson's correlation coefficient, r = 0.566; P < .001). However, there was no significant difference of PDGFRalpha expression between 2 grades of tumors. CONCLUSIONS: The results of this study showed that MIB-1 LI is a rapid and cost-effective modality for predicting tumor grade in oligodendrogliomas. Immunohistochemistry for PDGF was found to be useful in differentiating various grades of oligodendroglioma, and therefore, it may be involved in tumor cell proliferation and malignant transformation. Platelet-derived growth factor receptor alpha, although expressed in oligodendroglial neoplasms, was not found to be useful in predicting tumor grade. PMID- 19559930 TI - Stereoscopic virtual reality simulation for microsurgical excision of cerebral arteriovenous malformation: case illustrations. AB - BACKGROUND: Use of virtual reality planning and rehearsal for planning and training of cerebral aneurysm clipping was recently reported. We aimed to illustrate its applications in preoperative planning and training for cerebral AVM excision. CASE DESCRIPTION: We illustrated the application of preoperative rehearsal for excision of cerebral AVM for 2 patients in a stereoscopic virtual reality environment. One patient was a 44-year-old lady with a right posterior temporal AVM, and 1 patient was a 23-year-old lady with a left frontal AVM. Through Dextroscope, an anatomical understanding of arterial feeders, nidus, and draining veins in relationship to surrounding cerebral cortex was obtained. It allowed one to see the exposure with the different angles of visualization, similar to what happened under the operative microscope. CONCLUSION: Dextroscopic virtual reality stimulation provided an illustrated preoperative planning and training for excision of cerebral AVM. PMID- 19559933 TI - Neurosurgical training: more hours needed or a new learning culture? PMID- 19559938 TI - Re: "Validity of percutaneous controlled radiofrequency thermocoagulation in the treatment of isolated third division trigeminal neuralgia" by Fraioli et al. PMID- 19559939 TI - Authorship. PMID- 19559941 TI - Response to the editorial "Why we misunderstand the young generations" by Dr James Ausman. PMID- 19559942 TI - Single-port laparoscopic surgery: is a single incision the next frontier in minimally invasive gynecologic surgery? PMID- 19559947 TI - Nature. PMID- 19559948 TI - Dr Ross McIntire, otolaryngologist, and his care of President Franklin D. Roosevelt. AB - The role that otolaryngologist Ross McIntire, MD, played in the care of Franklin Delano Roosevelt, 32nd President of the United States, was documented by reviewing primary source material pertaining to the relationship of McIntire and Roosevelt. This included material from various archives including the Franklin D. Roosevelt Presidential Library at Hyde Park, New York; United States National Archives; and numerous autobiographies and diaries. McIntire's belief in the value of confidentiality and to provide information only on a need-to-know basis is consistent with the strategy that he had devised earlier for protecting his patient's privacy. In the context of his time and his position, Dr McIntire served his patient and his country well by making appropriate medical and wise personal judgments. The career of Dr Ross T. McIntire, otolaryngologist and personal physician to the 32nd president of the United States, engenders a sense of honor to our profession. PMID- 19559943 TI - Development of a neutralization assay for Nipah virus using pseudotype particles. AB - Nipah virus (NiV) and Hendra virus (HeV) are zoonotic paramyxoviruses capable of causing severe disease in humans and animals. These viruses require biosafety level 4 (BSL-4) containment. Like other paramyxoviruses, the plaque reduction neutralization test (PRNT) can be used to detect antibodies to the surface glycoproteins, fusion (F) and attachment (G), and PRNT titers give an indication of protective immunity. Unfortunately, for NiV and HeV, the PRNT must be performed in BSL-4 containment and takes several days to complete. Thus, we have developed a neutralization assay using VSV pseudotype particles expressing the F and G proteins of NiV (pVSV-NiV-F/G) as target antigens. This rapid assay, which can be performed at BSL-2, was evaluated using serum samples from outbreak investigations and more than 300 serum samples from an experimental NiV vaccination study in swine. The results of the neutralization assays with pVSV NiV-F/G as antigen showed a good correlation with those of standard PRNT. Therefore, this new method has the potential to be a rapid and cost-effective diagnostic method, especially in locations that lack high containment facilities, and will provide a valuable tool for basic research and vaccine development. PMID- 19559949 TI - Role of vestibular testing in diagnosis of benign paroxysmal positional vertigo. AB - Diagnosis and treatment of benign paroxysmal positional vertigo (BPPV) is a mixture of empiricism of particle repositioning with the rationally-based knowledge obtained from clinical observations, histopathology, and neurophysiological experiments. The recently published clinical practice guideline on BPPV makes recommendations on the management of BPPV. One of the statements discourages the use of radiographic or vestibular testing, unless the diagnosis was uncertain or there were additional signs or symptoms unrelated to BPPV. The role of video-oculography in diagnosis and treatment of BPPV is argued, since vestibular testing has provided key relevant information to understand positional nystagmus in patients with BPPV. PMID- 19559951 TI - Effect of preoperative hearing level on success of stapes surgery. AB - OBJECTIVE: We evaluated functional results after stapedotomy in terms of the influence of preoperative hearing threshold. STUDY DESIGN: Case series and chart review. SUBJECTS AND METHODS: We classified 141 patients according to preoperative mean air-conduction threshold: 40 to 54 dB, group 1; 55 to 69 dB, group 2; and 70 to 89 dB, group 3. Speech discrimination scores also were noted. RESULTS: All groups showed significant improvement. The greatest increase in postoperative speech discrimination score was seen in group 3. Group 1 improved from moderate hearing loss (mean, 49 dB) to normal (mean, 24 dB); group 2, from moderately severe loss (mean, 61.2 dB) to mild (mean, 28 dB); and group 3, from severe loss (mean, 76.4 dB) to moderate (mean, 41.1 dB). CONCLUSION: Success in treating otosclerosis does not depend on extent of preoperative hearing loss. Patients with severe loss improved to moderate, increasing the benefit from use of a hearing aid. The patients attained good auditory function, improving their quality of life. PMID- 19559952 TI - Management of single-sided deafness with the bone-anchored hearing aid. AB - OBJECTIVES: The benefits of the bone-anchored hearing aid (BAHA) for rehabilitation of conductive and mixed hearing loss are well established. Recently, the BAHA was used to rehabilitate patients with single-sided deafness (SSD). In this study, the benefits of the BAHA in SSD are presented. STUDY DESIGN: Case series with planned data collection. SETTING: Tertiary referral center. SUBJECTS AND METHODS: Twenty-one consecutive adult patients with SSD underwent single-stage BAHA implantation on the side of deafness. Testing in sound field was performed using the hearing-in-noise test (HINT) in both unaided and aided conditions. Speech and noise signals were delivered through two speakers oriented in two test paradigms. The outcomes were expressed as signal-to noise (S/N) ratios. Subjective benefit analyses were determined through two questionnaires: the Abbreviated Profile of Hearing Aid Benefit (APHAB) and the Glasgow Hearing Aid Benefit Profile (GHABP). RESULTS: All subjects demonstrated significant improvement in speech reception thresholds with the HINT using the BAHA, especially with the 90/270 speaker paradigm, in which the mean improvement over the unaided condition was 5.5 dB SPL (range, 2.0-11.0 dB; P=0.00001). Qualitative subjective outcome measures demonstrated additional benefits. CONCLUSION: In SSD patients, the BAHA provides significant subjective benefits and improves speech understanding in noise. PMID- 19559953 TI - Cervical vestibular evoked myogenic potentials (cVEMPs) in patients with superior canal dehiscence syndrome (SCDS). AB - OBJECTIVE: To determine the usefulness of both amplitude and threshold data from tone-burst cervical vestibular evoked myogenic potential (cVEMP) testing for the evaluation of superior canal dehiscence syndrome (SCDS). STUDY DESIGN: Case series with chart review. SUBJECTS AND METHODS: Sixty-seven patients underwent cVEMP testing. We correlated mean tone burst cVEMP amplitude and threshold data with temporal bone CT findings. Patients were excluded for Meniere's disease, middle ear disease, or otologic surgery. RESULTS: Superior canal dehiscence patients had higher mean cVEMP amplitudes (SCDS 173.8 microV vs non-SCDS 69.7 microV, P=0.031) and lower mean thresholds (SCDS 72.8 dB nHL vs non-SCDS 80.9 dB nHL) at 500 Hz. CONCLUSION: Patients with SCDS have larger amplitudes and lower thresholds on cVEMP testing at 500 Hz. This study supports the utility of tone burst cVEMPs for the evaluation of SCDS and is one of few large single-center studies to establish normative data. PMID- 19559954 TI - Minimally invasive video-assisted thyroidectomy: a retrospective study over two years of experience. AB - OBJECTIVE: The minimally invasive video-assisted thyroidectomy technique has slowly gained acceptance. Previous studies have reported advantages of better cosmetic results and faster postoperative recovery. We report preliminary results from our single assistant technique over the initial two years of inception at an academic training center. METHODS: This study consists of a retrospective chart review of 172 cases between May 2005 and September 2007. All cases started as video-assisted thyroidectomy were included. Demographic, pre- and postoperative clinical data, imaging results, and hospital stay were collected. RESULTS: Acceptable data were available for 24 male and 148 female patients who underwent the video-assisted procedure. Five cases were converted to the conventional thyroidectomy. Of the 172 cases, 60 total thyroidectomies and 112 hemithyroidectomies were performed, with 37 cases of malignancy. The average hospitalization was 1.43 days with mean incision length was 3.51 cm, mean surgical time of 91.37 minutes, and mean blood loss of 31 cc. Transient voice problems were noted in 10 patients. CONCLUSION: We found statistically improved operative times, hospital stay, and blood loss the first 2 years with a low rate of temporary complications. It appears that minimally invasive video-assisted thyroidectomy is a safe and feasible option to standard thyroidectomy in selected patients. PMID- 19559955 TI - Minimally invasive radioguided parathyroidectomy performed for primary hyperparathyroidism. AB - OBJECTIVE: Parathyroid surgery is often challenging due to considerable variability in anatomy. Minimally invasive radioguided parathyroidectomy (MIRP) is a directed surgical approach that has been made possible by the advent of new imaging techniques, specifically sestamibi scanning. In this study, we review our experience and outcomes with MIRP in one of the largest case series reported in the literature. STUDY DESIGN: Case series with chart review. SETTING: Tertiary care center. SUBJECTS AND METHODS: Subjects were 305 patients who underwent MIRP in our institution between 1997 and 2007. Data including symptoms, preoperative and postoperative calcium levels, and PTH levels were collected. Analyses were performed using Excel AnalysisPak. RESULTS: MIRP in this series had a 100 percent rate of success in removing a hyperfunctional parathyroid gland. The mean preoperative calcium was 10.9 whereas the mean postoperative level was 9.8. There was a significant difference between preoperative and postoperative calcium levels (both ionized and total) (P<0.01). Mean preoperative and postoperative serum PTH levels were 138 and 50, respectively. PTH levels were therefore significantly lower postoperatively (P<0.01). Rapid PTH testing showed a similar pattern. There were no cases of recurrent laryngeal nerve palsy. CONCLUSION: MIRP is an effective method for removal of image-localized hyperfunctional parathyroid glands. PMID- 19559956 TI - Surgical treatment of dysthyroid orbitopathy. AB - OBJECTIVES: Orbital decompression for dysthyroid orbitopathy may be performed by open or transnasal endoscopic approaches; however, criteria for the selection of the appropriate surgical technique have not been well-defined. Our goal was to compare the surgical outcomes of orbital decompression techniques employed by the otolaryngology and ophthalmology services at a single institution, so as to clarify the indications and develop a rationale for optimal management. STUDY DESIGN: Case series with chart review. SETTING: Tertiary care academic medical center. SUBJECTS AND METHODS: Orbital decompressions on 112 orbits of 69 patients from 2001 to 2008 at a tertiary care academic medical center. RESULTS: The majority (83%) of orbital decompressions were performed primarily for proptosis, whereas the remainder was indicated for dysthyroid optic neuropathy. Maximum reduction of proptosis was achieved with an endoscopic medial and inferior orbital wall decompression combined with a lateral orbitotomy with fat removal. The average reduction in proptosis was 7.4 mm+/-2.3 mm (primary surgery for proptosis). The endoscopic approach alone provided the least average proptosis reduction but was highly successful in the treatment of dysthyroid optic neuropathy. CONCLUSIONS: A three-wall decompression using a combined endoscopic and external approach provides the greatest amount of decompression. The endoscopic approach as a single modality is best suited for patients with mild proptosis and for patients with dysthyroid optic neuropathy. Cooperation between otolaryngology and ophthalmology achieves the best care for patients with thyroid eye disease who require surgical treatment. PMID- 19559957 TI - Revision endoscopic orbital decompression in the management of Graves' orbitopathy. AB - OBJECTIVE: Endoscopic orbital decompression has proved to be an effective treatment for Graves' orbitopathy. In select patients, however, persistent or recurrent orbital symptoms necessitate additional therapy. The objective of this study is to determine the safety and effectiveness of revision endoscopic orbital decompression in patients with refractory Graves' orbitopathy. STUDY DESIGN: Case control series. SETTING: Academic medical center. METHODS: The study population consisted of 10 patients with Graves' orbitopathy who underwent 13 revision endoscopic orbital decompressions (three bilateral cases) between 1991 and 2008. Outcome measures, including reduction in proptosis, improvement in visual acuity, and complication rates, were compared with a control cohort of 10 consecutive patients (16 orbits) who underwent primary endoscopic decompression during the same time period. RESULTS: Indications for revision decompression included exposure keratopathy (n=8, 62%), optic neuropathy (n=3, 23%), and gaze restriction (n=2, 15%). There were no intraoperative complications. Mean reduction in proptosis was 1.4 mm less for patients who underwent revision decompression compared with primary cases (mean decompression 3.6+/-1.0 mm vs 5.0+/-2.1 mm, respectively), although this difference was not statistically significant (P=0.13). Visual acuity improved in 62 percent of revision cases, compared with 20 percent of primary cases (P=0.09). Rates for postoperative complications, which included sinusitis and frontal mucocele formation, were also similar between revision and primary decompression groups (38% vs 13% respectively, P=0.17). CONCLUSIONS: This report is the first to describe the endoscopic technique for revision orbital decompression. It appears to be a safe and effective procedure for the treatment of refractory orbitopathy in patients with Graves' disease. PMID- 19559958 TI - Molecular markers of micrometastasis in oral cavity carcinomas. AB - OBJECTIVE: To examine the expression of candidate markers for micrometastasis. STUDY DESIGN: Cross-sectional analysis of subjects with oral cavity carcinomas who underwent sentinel lymph node biopsy (SLNB) and subsequent immunohistochemical (IHC) analysis. SUBJECTS AND METHODS: Two groups were identified based on SLNB status: negative SLNB (19/30) and positive SLNB (11/30). Specimens underwent IHC using conjugated monoclonal antibodies for membrane type 1 matrix metalloproteinase (MT1-MMP), CD44, focal adhesion kinase-1, and E cadherin. Staining results were evaluated to determine if a particular marker was associated with SLNB status or other histopathologic prognosticators. RESULTS: For MT1-MMP, 21 percent (3/14) of evaluable specimens stained positively in the SLNB(-) group and 67 percent (4/6) stained positively in the SLNB(+) group (P=0.12). No statistically significant association was seen between any marker's staining pattern and SLNB status alone. When MT1-MMP staining was evaluated in tumors with SLNB(+) or perineural invasion (PNI) present on histopathology, six of nine specimens (67%) stained positively for MT1-MMP, vs one of 11 (9%) in specimens lacking either negative prognosticator (P=0.016, RR=7.33). CONCLUSION: Preliminary results suggest that MT1-MMP positivity in primary tumor specimens may identify aggressive tumor types, evidenced by the presence of micrometastasis or PNI. PMID- 19559960 TI - Functional and oncologic results following transoral laser microsurgical excision of base of tongue carcinoma. AB - OBJECTIVE: The study objective was to determine survival and functional results in patients who underwent transoral laser excision of base of tongue squamous cell carcinoma. STUDY DESIGN: This was a case series with chart review. SUBJECTS AND METHODS: Patients with biopsy-proven base of tongue squamous cell carcinoma primarily treated with transoral laser excision and followed for at least 24 months were included. Disease-specific and overall survival data were determined. Quality-of-life data were obtained using the University of Washington Quality of Life instrument. RESULTS: Of the 71 patients treated, one (1.4%), nine (12.7%), seven (9.9%), and 54 (76.1%) were stage I, II, III, and IV, respectively, at diagnosis. At 24 months, overall survival was 90 percent; disease-specific survival was 94 percent. Locoregional recurrence occurred in 10 percent. Given the low number of deaths, no significance was detectable between survival and gender, overall stage, tumor size, nodal status, or adjuvant therapy. Quality-of life data, obtained for 46 patients, revealed the majority of patients had mild or no pain, minimally impaired to normal swallowing, and normal speech. CONCLUSION: Transoral laser excision offers an acceptable treatment approach in terms of survival and quality of life for patients with base of tongue squamous cell carcinoma. PMID- 19559959 TI - Management of the N0 neck in moderately advanced squamous carcinoma of the larynx. AB - OBJECTIVE: To assess the oncological efficacy of selective neck dissection (SND) in patients with T3-4 N0 laryngeal squamous carcinoma. SUBJECTS AND METHODS: A total of 327 patients underwent 654 neck dissections; each side of the neck was individually evaluated. RESULTS: Three percent of patients who had SND developed regional recurrence (RR) in comparison with 11.7 percent of patients who underwent modified radical neck dissection (MRND) (P=0.005). Only 3 (0.9%) patients developed RR outside the field of SND. The presence of extracapsular extension (P=0.002) in node-positive (pN+) group and of microvascular invasion (P=0.007), together with the type of neck dissection (ND) (P=0.0003) in node negative (pN0) group had statistical impact on RR. The development of RR significantly affected disease-specific survival (P=0.0001). Equivalent rates of RR were found in pN+ (2.6%) or pN0 (3.2%) patients treated with SND (P=0.98) as well as in pN+ patients who underwent SND (2.6%) or MRND (4.7%) (P=0.85). CONCLUSION: This study confirmed the adequacy of SND as a satisfactory staging and therapeutic procedure, and suggests its use in the treatment of limited node positive (N+) neck. PMID- 19559961 TI - Modified frontolateral partial laryngectomy without tracheotomy. AB - OBJECTIVE: To investigate the feasibility of modified frontolateral partial laryngectomy without tracheotomy for patients with early-stage laryngeal cancer or dysplasia of the true vocal cord. STUDY DESIGN: After frontolateral partial laryngectomy, the inner side of the sternohyoid muscle was drawn into the laryngeal lumen to suture it to the incisal margin of the uninjured side and to the false vocal cord on the side of the lesion to ensure the safety without tracheotomy. A reverted sternohyoid fascial flap was used to cover the anterior area to form a new laryngeal lumen in the shape of a ladder. SUBJECTS AND METHODS: A total of 65 patients with early glottic carcinomas or severe dysplasia of the true vocal cord were treated with modified frontolateral partial laryngectomy without tracheotomy. Ipsilateral false vocal cord flaps and cervical skin flaps were used in 63 patients and two patients respectively to reconstruct the defect. RESULTS: Deglutition and phonation were fully recovered after 7 to 10 days postsurgery. The only postoperative complication was subcutaneous emphysema noted in nine patients. The 1-, 3-, and 5-year survival rates were all 100 percent. CONCLUSION: Modified frontolateral partial laryngectomy without tracheostomy is an effective surgical method for early glottic carcinomas and severe dysplasia of the true vocal cord. PMID- 19559962 TI - Histopathological changes of rat larynx mucosa with exposure to chronic thinner inhalation. AB - OBJECTIVE: Histopathological changes in nasal mucosa, trachea, and pulmonary system with exposure to chronic thinner inhalation have been studied in the literature. However, the possible changes in larynx mucosa, which is a part of the upper airway tract, have not been studied yet. The aim of this study is to determine the histopathological changes of rat larynx mucosa with exposure to chronic thinner inhalation. STUDY DESIGN AND SETTING: Randomized trial. The study was conducted at the animal care facility of Haydarpasa Numune Education and Research Hospital. SUBJECTS AND METHODS: Fifty-one Sprague-Dawley rats were used throughout the experiment. Four groups of rats inhaled thinner in a glass cage for 2, 4, 8, and 12 weeks respectively. Seven rats inhaled only the air in the room as the control group. RESULTS: The comparison of inflammation and exocytosis in the control and 2 week groups revealed no significant difference (P>0.05), but from the beginning of 4 weeks of thinner inhalation, statistically significant differences were observed (P<0.05). From the beginning of 8 weeks of thinner inhalation, statistically significant differences were observed in larynx mucosa when we assessed metaplasia and cilia loss distribution among groups (P<0.05). CONCLUSION: On the basis of histopathological evaluations, it was shown that the harmful effect of inhalation of thinner in high concentrations to larynx mucosa is similar to the effect on other organs of the respiratory system. PMID- 19559963 TI - Persistent pediatric obstructive sleep apnea and lingual tonsillectomy. AB - OBJECTIVE: To describe a new method and the indications for lingual tonsillectomy with endoscopy and coblation, and to document its utility for treating children with persistent obstructive sleep apnea after previous tonsillectomy and adenoidectomy. STUDY DESIGN AND SETTING: Case series with chart review in a tertiary pediatric medical center. SUBJECTS AND METHODS: Twenty-six patients aged 3 to 20 met the inclusion criteria of polysomnography-proven persistent obstructive sleep apnea after tonsillectomy and adenoidectomy, as well as diagnosis of lingual tonsillar hypertrophy made by flexible fiberoptic sleep endoscopy. Endoscopic-assisted coblation lingual tonsillectomies were performed between June 2005 and January 2008. Preoperative and postoperative nocturnal polysomnogram data were paired and analyzed statistically. RESULTS: Statistically significant reductions in the respiratory distress index (RDI) were seen when preoperative and postoperative data were compared (mean, 14.7 vs 8.1). There were similar reductions in the number of obstructive apneas and hypopneas. The mean minimum O2 saturation did not change. Two patients in this series developed adhesions between the epiglottis and tongue base; there appeared to be no consequence for airway or feeding issues. CONCLUSION: Endoscopic-assisted coblation lingual tonsillectomy is an effective technique for the treatment of lingual tonsillar hypertrophy causing persistent obstructive sleep apnea in some children. PMID- 19559964 TI - Sensorineural hearing loss in patients with cystic fibrosis. AB - OBJECTIVE: To determine the prevalence of sensorineural hearing loss (SNHL) in cystic fibrosis (CF) patients and its relationship to antibiotic use. STUDY DESIGN: Case series with chart review. SETTING: Tertiary care pediatric hospital. SUBJECTS AND METHODS: We reviewed the medical records of CF patients seen in our children's hospital between March 1994 and December 2007. Data collected included patient demographics, audiograms, tympanograms, genotype, and use of potentially ototoxic antibiotics. RESULTS: Seven of 50 (14%) patients had SNHL. Three percent of patients who received 10 courses (P<0.01). No patients who received five or fewer courses of nasal irrigation with aminoglycosides had SNHL versus 23 percent of those who received more than five courses (P<0.05). Nine percent of patients who received five or fewer courses of macrolides had SNHL versus 60 percent of those who received more than five courses (P=0.079). CONCLUSION: CF patients receiving aminoglycosides are at high risk for developing SNHL. PMID- 19559965 TI - Immunological investigation in the adenoid tissues from children with chronic rhinosinusitis. AB - OBJECTIVE: Chronic rhinosinusitis (CRS) is characterized by persistent inflammation and tissue remodeling of the nasal mucosa. Adenoidectomy is an effective surgical treatment in pediatric CRS. To evaluate the effect of pediatric CRS on the severity and characteristics of adenoid inflammation, the authors evaluated the expressions of inflammatory cell activation markers and tissue remodeling in adenoid tissues associated with cytokines tissue-remodeling associated cytokines in adenoid tissues. STUDY DESIGN AND SETTING: A prospective controlled study on 40 pediatric patients admitting for adenotonsillectomy. SUBJECTS AND METHODS: Immunoassays were performed on adenoid tissues homogenates from 16 children with CRS and from 24 children without CRS to quantify the levels of inflammatory cell activation markers, such as soluble interleukin (IL)-2 receptor (sIL-2R), soluble CD23 (sCD23), IL-6, eosinophilic cationic protein (ECP), and tryptase, and the levels of cytokines associated with tissue remodeling, such as transforming growth factor (TGF)-beta1, matrix metalloproteinase (MMP) 2 and 9, and tissue inhibitor of metalloproteinase (TIMP) 1. RESULTS: The mean levels (the ratio to albumin level) of sIL-2R, TGF-beta1, MMP-2, MMP-9, and TIMP-1 were significantly higher in adenoid tissues of patients with CRS (27.31+/-30.32, 4894.65+/-2388.77, 500.13+/-604.59, and 23.06+/-10.37, respectively) than those without it (16.27+/-10.93, 2635.51+/-1448.63, 120.87+/ 321.50, 16.74+/-11.10, and 7.39+/-3.12, respectively; all P<0.05). Regarding the severity of CRS, ECP level was significantly higher in patients with severe CRS than in those with mild to moderate CRS (P=0.033). CONCLUSIONS: Adenoid tissues in pediatric CRS patients had higher levels of tissue-remodeling-associated cytokines, which may explain the relationship between pediatric CRS and adenoid inflammation. PMID- 19559966 TI - Local production of antigen-specific IgE in different anatomic subsites of allergic fungal rhinosinusitis patients. AB - OBJECTIVE: Local production of antigen-specific IgE in allergic fungal rhinosinusitis (AFRS) is likely integral to the expression of allergy. This study examines if there are anatomic variations in local IgE expression or if variations among fungal and nonfungal IgE exist. STUDY DESIGN: Cross-sectional study. SETTING: Tertiary medical center. SUBJECTS AND METHODS: Specimens from 11 AFRS, 8 chronic rhinosinusitis without nasal polyps (CRSsNP), and 9 control patients underwent immunohistochemical localization for IgE and evaluation for antigen-specific IgE by ImmunoCAP testing. RESULTS: Inferior turbinate (IT) epithelium had greater IgE staining in AFRS than control (P=0.013) and CRSsNP (P=0.002). A significant difference was also found at the IT subepithelial level for AFRS compared with controls (P=0.001) and CRSsNP (P<0.001). Within AFRS, IgE staining was increased in the subepithelium compared to epithelium (P=0.003). ImmunoCAP analysis on IT tissue from AFRS and controls demonstrated increased antigen-specific IgE for 5 of 14 antigens (P<0.05) and total IgE (P<0.001). There were no significant anatomic differences between IT and sinus IgE staining. CONCLUSION: More fungal and nonfungal IgE is expressed in IT and sinus tissues of AFRS patients, as compared with control and CRSsNP patients. PMID- 19559967 TI - The prevalence of atopic symptoms in children with otitis media with effusion. AB - OBJECTIVE: To determine the prevalence of allergic symptoms in children with otitis media with effusion (OME). STUDY DESIGN: A validated questionnaire from the International Study of Asthma and Allergies in Childhood was used to determine the prevalence of allergic symptoms in children. The questionnaire was completed by the parents of children with OME undergoing ventilation tube insertion, and the results were compared with a large reference group of school children of the same age. SUBJECTS AND METHODS: Children aged 6 or 7 years old with OME confirmed intraoperatively during ventilation tube insertion between 2001 and 2005 (n=89). The prevalence of allergic symptoms and nasal symptoms in children with OME was compared with an age-matched reference group. RESULTS: There was no difference in the prevalence of allergic symptoms suggesting rhinoconjunctivitis, asthma, or eczema between the OME and reference group. The prevalence of nasal symptoms, however, was greater in the children with OME than in the reference group 38.2 percent versus 23.5 percent (odds ratio=2.01; 95% confidence interval, 1.30-3.10; P<0.001). CONCLUSION: The prevalence of allergic symptoms was similar in 6- to 7-year-old children with OME and the reference group, suggesting a limited effect of allergy in the pathogenesis of OME in this age group. Nasal symptoms were more common in the OME group, which may reflect a higher prevalence of adenoidal hyperplasia. PMID- 19559969 TI - Effectiveness of honey on Staphylococcus aureus and Pseudomonas aeruginosa biofilms. AB - OBJECTIVES: Biofilms formed by Pseudomonas aeruginosa (PA) and Staphylococcus aureus (SA) have been shown to be an important factor in the pathophysiology of chronic rhinosinusitis (CRS). As well, honey has been used as an effective topical antimicrobial agent for years. Our objective is to determine the in vitro effect of honey against biofilms produced by PA and SA. STUDY DESIGN: In vitro testing of honey against bacterial biofilms. METHODS: We used a previously established biofilm model to assess antibacterial activity of honey against 11 methicillin-susceptible SA (MSSA), 11 methicillin-resistant SA (MRSA), and 11 PA isolates. Honeys were tested against both planktonic and biofilm-grown bacteria. RESULTS: Honey was effective in killing 100 percent of the isolates in the planktonic form. The bactericidal rates for the Sidr and Manuka honeys against MSSA, MRSA, and PA biofilms were 63-82 percent, 73-63 percent, and 91-91 percent, respectively. These rates were significantly higher (P<0.001) than those seen with single antibiotics commonly used against SA. CONCLUSION: Honey, which is a natural, nontoxic, and inexpensive product, is effective in killing SA and PA bacterial biofilms. This intriguing observation may have important clinical implications and could lead to a new approach for treating refractory CRS. PMID- 19559968 TI - Estrogen receptor overexpression in malignant minor salivary gland tumors of the sinonasal tract. AB - OBJECTIVES: To identify the expression of estrogen receptors in malignant minor salivary gland tumors of the sinonasal tract. STUDY DESIGN: Case series with chart review. SUBJECTS AND METHODS: A retrospective review of a pathology database identified 17 patients with malignant salivary gland tumors between December 1987 and January 2006. Clinicopathologic data were collected, and immunohistochemical staining for estrogen receptor alpha and beta was performed. RESULTS: Among these malignant tumors, adenoid cystic carcinoma was the predominant histologic type. In addition to epistaxis and nasal obstruction, the headache, facial, or ocular symptoms were also commonly noted in this cohort. Seventy-five percent of cases of adenoid cystic carcinoma were positive for estrogen receptor alpha. In contrast, only 17 percent of cases of adenoid cystic carcinoma were positive for estrogen receptor beta. CONCLUSION: Malignant minor salivary gland tumors of the sinonasal tract are rare disease entities. In the present series, adenoid cystic carcinoma was the most common form of tumor, but the prognosis was poor. Most of the cases were positive for expression of estrogen receptor alpha, which suggests that hormone therapy may have a role in the management of certain minor salivary gland tumors of the paranasal sinus and nasal cavity. PMID- 19559970 TI - Effects of a tongue-holding maneuver during swallowing evaluated by high resolution manometry. AB - OBJECTIVE: This study aimed to investigate the effects of a tongue-holding maneuver (THM) during swallowing using a novel high-resolution manometry (HRM) system. STUDY DESIGN: Case series with planned data collection. MATERIALS AND METHODS: Thirty-three asymptomatic Japanese adults were studied. A solid-state HRM assembly with 36 circumferential sensors spaced 1-cm apart was positioned to record pressures from the velopharynx to the upper esophagus at rest and during swallowing. The maximum values of the dry swallowing pressures at the velopharynx, mesohypopharynx, upper esophageal sphincter (UES), and distance from the nostril to each point of maximum values with and without the THM were measured. RESULTS: The distance from the nostril to the UES was statistically shorter when swallowing with the THM than without the THM (paired t test, P=0.009). The maximum pressure at UES was greater when swallowing with the THM than without the THM, although there was no statistically significant difference (paired t test, P=0.051). There was no difference in the pressures and the distance between swallowing with or without the THM at any other site. CONCLUSIONS: These findings suggest that the THM may not have a potential to facilitate compensatory swallowing power when swallowing. PMID- 19559971 TI - Diagnosis and treatment of deep neck space abscesses. AB - OBJECTIVE: To review our experience with deep neck abscesses and identify unique trends in our patient population. STUDY DESIGN: Case series with chart review. SUBJECTS AND METHODS: Evaluation of patients with deep neck space abscesses between 2001 and 2006. Peritonsillar abscess, superficial craniocervical infection, and salivary gland infections were excluded from selection of study population. A total of 106 cases were reviewed. RESULTS: Dental infections were the most common cause of deep neck abscesses (49.1%). Comorbidities included substance abuse (53.7%), psychiatric illness (10.4%), hypertension (9.4%), head and neck cancer (6.6%), and diabetes mellitus (5.7%). All patients received systemic antibiotics, eight patients required tracheotomy, 85 patients underwent surgical drainage in the operating room, and 11 had bedside drainage. Median and lower quartile of time in hospital was 2 and 3 days, respectively, whereas upper quartile was 4 days (range, 1 to 27 days). Patients with comorbidities or concurrent illness tended to stay longer (P<0.05, Mann-Whitney test). There were six complications and no mortality. CONCLUSION: Substance abuse and poor orodental hygiene are important predisposing factors to deep neck abscesses. Appropriate management continues to favor a combination of early surgical drainage and systemic antibiotics. PMID- 19559972 TI - Real-time PCR detection of Helicobacter pylori and virulence-associated cagA in nasal polyps and laryngeal disorders. AB - OBJECTIVE: To identify Helicobacter pylori and major virulence factor, cagA, in patients with laryngeal diseases and nasal polyps. STUDY DESIGN: Cross-sectional study with planned data collection. SETTING: The study was performed on fresh tissue samples from patients with 32 nasal polyps, 29 normal nasal mucosa, and 27 laryngeal diseases presenting to the Otolaryngology-Head and Neck Surgery department of a major military hospital in Istanbul, Turkey. SUBJECTS AND METHODS: Tissue specimens were evaluated by in-house polymerase chain reaction (PCR) and real-time PCR for bacterial DNA and by real-time PCR for cagA. The impact of commercial and in-house DNA extraction methods was also evaluated. RESULTS: H pylori DNA was detected only by real-time PCR in 59.4 percent of nasal polyps, 70.4 percent of nasal mucosa samples, and 58.6 percent of larynx samples. cagA was identified in 78.9, 89.5, and 82.4 percent of positive polyp, nasal mucosa, and larynx samples, respectively. No statistically significant differences were observed between groups. DNA purification methods were equally effective. CONCLUSION: H pylori DNA is present in nasal polyp and larynx tissues as well as normal nasal mucosa, as detected by a sensitive real-time PCR assay. cagA-positive strains dominate in all groups. PMID- 19559973 TI - Prevalence of depression and antidepressant use in an otolaryngology patient population. AB - OBJECTIVE: To determine the prevalence of depressive diagnoses and antidepressant use in various subsets of otolaryngology patients. STUDY DESIGN: Academic practice EMR database query. SUBJECTS AND METHODS: Over 12,000 consecutive otolaryngology patients were analyzed by primary diagnosis and in three groups: Group 1 (tinnitus, vertigo), Group 2 (rhinosinusitis), and Group 3 (sleep apnea). The number with depressive diagnoses and/or those taking antidepressants was determined. RESULTS: The prevalence of any depressive diagnosis was 11 percent, and 18 percent had been prescribed antidepressants. The prevalence of depression was 13 percent, 10 percent, and 14 percent for Groups 1, 2, and 3, respectively. Antidepressants had been prescribed in 21 percent, 15 percent, and 28 percent, respectively. The primary-encounter diagnosis with the highest incidence of depression (21%) and antidepressant use (46%) was sleep apnea with insomnia. CONCLUSION: Otolaryngology patients may exhibit greater prevalence of depression than is observed in the general population (7.3%). Highest prevalence was found in patients with inner ear disease and sleep apnea. Depression may be a cause of significant comorbidity in patients with chronic otolaryngic conditions. PMID- 19559974 TI - Bilateral simultaneous free gracilis muscle transfer: a realistic option in management of bilateral facial paralysis. AB - OBJECTIVES: Bilateral facial paralysis patients have oral incompetence, poor articulation, and dental caries. This problem is frequently addressed by performing staged gracilis transplants without specific oral sphincter reconstruction. The purpose of this study is to describe the technique of bilateral simultaneous free gracilis muscle transfer with oral sphincter reconstruction, for one-stage facial reanimation in patients with bilateral facial paralysis. STUDY DESIGN: Case series. METHODS/RESULTS: One-stage bilateral gracilis transfer was performed in three patients with bilateral facial paralysis. Muscle transplants produced a meaningful smile in all transferred muscles. All patients reported improved speech and decreased drooling. CONCLUSIONS: Bilateral gracilis transplants with reconstruction of the oral commissure is a realistic option in management of the bilaterally paralyzed face. In this small series, it appears to improve oral competence, restore smiling, and contribute favorably to lower lip support. This technique has been made more feasible because the operative time for free tissue transfer has continuously declined with the use of two-team surgery, improved microsurgical techniques, and the advent of venous coupling devices. PMID- 19559975 TI - Use of an oral sensory feedback device in the management of jaw-opening dystonia. PMID- 19559976 TI - Bilateral hypopharyngeal diverticulum. PMID- 19559977 TI - Facial nerve schwannoma. PMID- 19559978 TI - A nose peg: a possible simple solution for musicians with velopharyngeal incompetence. PMID- 19559979 TI - Henoch-Schonlein purpura after adenotonsillectomy. PMID- 19559980 TI - A contraindication to routine neck manipulation examinations in patients with acute vertigo: vertebral artery dissection. PMID- 19559981 TI - Auricular tophi as the initial presentation of gout. PMID- 19559982 TI - The role of tonsillectomy in reducing recurrent pharyngitis: a systematic review. PMID- 19559984 TI - Distribution of tryptophan hydroxylase-immunoreactive neurons in the brainstem and diencephalon of the pigeon (Columba livia). AB - The distribution of tryptophan hydroxylase (TPH)-containing perikarya and processes in the brainstem and diencephalon of the pigeon (Columba livia) were investigated using single-labeling chromogenic and double-labeling fluorescence immunohistochemical methods for TPH and 5-HT. TPH-immunoreactive (TPH-ir) perikarya were seen extending from the caudal medulla to mid-hypothalamic levels, located in brainstem regions previously described as containing 5-HT-ir somata. Brainstem TPH-ir cell clusters (the midline raphe, and the dorsolateral and ventrolateral serotonergic cell groups) and the circumventricular cerebrospinal fluid-contacting neurons in the taenia choroidea (in the caudal brainstem), recessus infundibuli and paraventricular organ (in the hypothalamus) were shown to co-express 5-HT immunoreactivity. However, heavily labeled TPH-ir cell clusters were observed in the nucleus premamillaris (PMM), in the stratum cellulare internum (SCI), in the nucleus paraventricularis magnocellularis (PVN) and in the medial border of the nucleus dorsomedialis anterior thalami (DMA). Double-labeling experiments indicated that none of these medial hypothalamic TPH ir cells were immunoreactive to 5-HT. These cells correspond to dopamine- and melatonin-containing neurons previously found in the avian hypothalamus, and appear to be comparable to the mammalian TPH-ir hypothalamic A11-A13 catecholaminergic somata, suggesting that they may be a conserved attribute in the amniote medial hypothalamus. PMID- 19559985 TI - Pre-treatment with lidocaine suppresses ectopic discharges and attenuates neuropeptide Y and c-Fos expressions in the rat cuneate nucleus following median nerve transection. AB - Following peripheral nerve injury, lidocaine application has been demonstrated to suppress injury discharges. However, there is very little information about the effects of lidocaine pre-treatment. The aim of the present study was to examine the effects of pre-treatment with lidocaine on injury discharges of the nerve, and neuropeptide Y (NPY) and c-Fos expression in the cuneate nucleus (CN) after median nerve transection (MNT). Rats received either saline or 1%, 5%, or 10% lidocaine applied topically to the median nerve before nerve transection. Electrophysiological recording was used to examine the changes in injury discharges of the nerve at post-injection, transection, pre- and post-electrical stimulation stages in the different groups. Sequential immunohistochemistry was also used to identify the number of NPY-like immunoreactive (NPY-LI) fibers and c Fos-LI cells in the corresponding CN. An increasing frequency of injury discharges was observed at all stages in the pre-saline group, which were suppressed by lidocaine pre-treatment in a dose-dependent manner. Lidocaine pre treatment also attenuated the number of injury-induced NPY-LI fibers and c-Fos-LI neurons within the CN in a dose-dependent manner. Furthermore, expression of c Fos-LI neurons in the CN was significantly reduced by an NPY receptor antagonist, indicating that NPY modulated c-Fos expression following MNT. These data suggest that preventing injury discharges with lidocaine pre-treatment can effectively attenuate central sensitization following MNT. PMID- 19559986 TI - Nuclear organization and morphology of cholinergic, putative catecholaminergic and serotonergic neurons in the brain of the rock hyrax, Procavia capensis. AB - The nuclear subdivisions of the cholinergic, putative catecholaminergic and serotonergic systems within the brain of the rock hyrax (Procavia capensis) were identified following immunohistochemistry for acetylcholinesterase, tyrosine hydroxylase and serotonin. The aim of the present study was to investigate possible differences in the complement of nuclear subdivisions of these systems by comparing those of the rock hyrax to published studies of other mammals. The rock hyrax belongs to the order Hyracoidea and forms part of the Afroplacentalia mammalian cohort. For the most part, the nuclear organization of these three systems closely resembled that described for many other mammalian species. The nuclear organization of the serotonergic system was identical to that seen in all eutherian mammals. The nuclear organization of the putative catecholaminergic system was very similar to that seen in rodents except for the lack of a C3 nucleus and the compact division of the locus coeruleus (A6c). In addition, the diffuse locus coeruleus (A6d) appeared to contain very few tyrosine hydroxylase immunoreactive (TH+) neurons. The cholinergic system showed many features in common with that seen in both rodents and primates; however, there were three differences of note: (1) cholinergic neurons were observed in the anterior nuclei of the dorsal thalamus; (2) cholinergic parvocellular nerve cells, probably representing interneurons, forming subdivisions of the laterodorsal and pedunculopontine tegmental nuclei were observed at the midbrain/pons interface; and (3) a large number of cholinergic nerve cells in the periventricular grey of the medulla oblongata were observed. Thus, while there are many similarities to other mammalian species, the nuclear organization of these systems in the rock hyrax shows specific differences to what has been observed previously in other mammals. These differences are discussed in both a functional and phylogenetic perspective. PMID- 19559987 TI - Focus on overweight and obesity prevention and management. PMID- 19559988 TI - Pharmacotherapy and surgery treatment for the severely obese adolescent. PMID- 19559989 TI - Do parents accurately perceive their child's weight status? AB - INTRODUCTION: Few studies have evaluated the accuracy of parental perceptions of their child's weight status. METHODS: A cross-sectional sample of children aged 5 to 12 years and their parents (n = 576 parent-child pairs) was enrolled from four schools. Child height and weight were measured. The parents classified their child on Likert scales ranging from "extremely overweight" to "extremely underweight." Parental perceptions were compared with their child's weight status according to body mass index (BMI) age-gender percentiles. Fisher-Halton-Freeman tests, chi(2), and logistic regression were used to compare demographic factors between parents who inaccurately estimated and those who accurately estimated child weight status. RESULTS: Misclassification occurred 25% of the time (95% confidence interval: 21.4-28.5). All parents of children with a BMI greater than or equal to the 95th percentile classified their child in a category other than "extremely overweight," and 75% of children with a BMI from the 85th to less than the 95th percentile were misclassified as "about right" or "underweight." Boys were more likely to be misclassified than were girls (29% vs 21%, P = .03). CONCLUSIONS: The majority of parents of obese and overweight children underestimate their child's weight status. Parents of boys are more likely to perceive their child's weight incorrectly. PMID- 19559990 TI - Changes in nurse practitioners' knowledge and behaviors following brief training on the healthy eating and activity together (HEAT) guidelines. AB - INTRODUCTION: Primary care providers, particularly pediatric nurse practitioners, are an integral force involved in tackling the obesity epidemic among youth. The majority of nurse practitioners, however, report low proficiency regarding their ability to adequately prevent and treat pediatric overweight. In response, the National Association of Pediatric Nurse Practitioners (NAPNAP) developed the evidence-based Healthy Eating and Activity Together (HEAT) Clinical Practice Guideline (CPG) to improve provider behavior and efficacy. METHOD: Thirty-five nurse practitioners attending the NAPNAP Annual Conference participated in an intensive 4-hour HEAT CPG training session. Pre-training and post-training data were collected on provider knowledge, practice behaviors, and barriers in relation to the prevention of overweight among youth. RESULTS: Post-training results revealed significant improvements in (a) practitioner knowledge (assessment of patient growth, family history, psychosocial functioning, nutrition, and physical activity); (b) practitioners' intent to improve behavior (i.e., increased intent to use behavior modification and counseling aimed at patient and family behavior change); and (c) practitioners' report of increased confidence in ability to address barriers. DISCUSSION: Study findings demonstrate preliminary support for the HEAT CPG as an effective tool aimed at helping providers to improve their ability to maintain patients' healthy weight. Future research is needed to verify the effects of HEAT CPG on long-term improvements in care. PMID- 19559991 TI - Pediatric nurse practitioners' assessment and management of childhood overweight/obesity: results from 1999 and 2005 cohort surveys. AB - INTRODUCTION: Recently, public and professional emphasis has been placed on addressing the increasing prevalence of childhood overweight. METHOD: This survey study was conducted with two cohorts of pediatric nurse practitioners (N = 413) to explore differences in self-reported practice skills over time. RESULTS: Significant improvements in assessment, screening, and laboratory evaluations were reported, although reduced adherence to recommended psychosocial assessments was noted. DISCUSSION: This study outlines self-reported barriers to effective childhood weight management. One support that participants requested was evidence based guidelines. Motivational interviewing may be an additional strategy to enhance provider skills to assess and manage challenging patient behavior change (e.g., dietary and activity changes). PMID- 19559992 TI - Integrative review of school-based childhood obesity prevention programs. AB - Childhood and adolescent overweight and obesity in the United States are rapidly becoming a major pediatric health concern. Schools are a critical part of the social environment that shape children's eating and physical activity patterns. While school-based programs may affect a student's perception of physical activity and the influence of dietary practices on overall health, the current variability in the theoretical underpinnings and methodological approaches used to implement these programs makes them difficult to evaluate for quality and evidence of effectiveness of the outcomes achieved across programs. The purpose of this study is to conduct an integrative research review using Cooper's framework to provide an overview of the degree of variability in the methodological approaches and theoretical frameworks of school-based obesity prevention programs that utilize one or more of the following interventions: dietary, physical activity, healthy lifestyle education, and/or parental involvement. This review will examine the variations in the duration of the interventions; whether the interventions were guided by the use of a theoretical framework; the strength of evidence supporting the studies; and whether the interventions demonstrated a reduction in body mass index or weight loss. A total of 16 articles were found based on the inclusion criteria for this study. Eight of the 16 studies (50%) applied theoretical frameworks. Significant variability was found in the duration of intervention between the studies analyzed. The range of duration was 5 weeks to 8 years, with an average of 16.8 months. Fourteen of 16 studies (88%) implemented dietary habit interventions. Fourteen of 16 studies (88%) implemented physical education programs. All 16 studies applied healthy lifestyle education. Nine of 16 studies integrated family involvement into the obesity intervention. Nine of 16 studies (56%) evaluated the effect of their intervention on body mass index. PMID- 19559993 TI - Gastrointestinal comorbidities in an obese child. PMID- 19559994 TI - Adolescent with history of hypercholesterolemia and ADHD: evidence-based practice. PMID- 19559995 TI - State legislators lead fight against childhood obesity. PMID- 19559996 TI - Circumventing communication barriers with Spanish-speaking patients regarding pediatric obesity. PMID- 19559998 TI - Society of Interventional Radiology clinical practice guidelines. PMID- 19559999 TI - Quality improvement guidelines for uterine artery embolization for symptomatic leiomyomata. PMID- 19560000 TI - Quality improvement guidelines for recording patient radiation dose in the medical record. PMID- 19560001 TI - Quality improvement guidelines for percutaneous management of acute limb ischemia. PMID- 19560002 TI - Quality improvement guidelines for transhepatic arterial chemoembolization, embolization, and chemotherapeutic infusion for hepatic malignancy. PMID- 19560003 TI - Quality improvement guidelines for the treatment of lower extremity deep vein thrombosis with use of endovascular thrombus removal. PMID- 19560004 TI - Recommendations for the implementation of joint commission guidelines for labeling medications. PMID- 19560006 TI - Guidelines for patient radiation dose management. PMID- 19560005 TI - Quality improvement guidelines for preventing wrong site, wrong procedure, and wrong person errors: application of the joint commission "universal protocol for preventing wrong site, wrong procedure, wrong person surgery" to the practice of interventional radiology. PMID- 19560007 TI - Interventional fluoroscopy: reducing radiation risks for patients and staff. PMID- 19560008 TI - The IR Radlex Project: an interventional radiology lexicon--a collaborative project of the Radiological Society of North America and the Society of Interventional Radiology. PMID- 19560010 TI - SIR/RSNA/CIRSE joint medical simulation task force strategic plan executive summary. PMID- 19560009 TI - Occupational health hazards in the interventional laboratory: time for a safer environment. AB - This document is a consensus statement by the major American societies of physicians who work in the interventional laboratory environment. It reviews available data on the prevalence of occupational health risks and summarizes ongoing epidemiologic studies designed to further elucidate these risks. Its purpose is to affirm that the interventional laboratory poses workplace hazards that must be acknowledged, better understood, and mitigated to the greatest extent possible. Vigorous efforts are advocated to reduce these hazards. Interventional physicians and their professional societies, working together with industry, should strive toward minimizing operator radiation exposure, eliminating the need for personal protective apparel, and ending the orthopedic and ergonomic consequences of the interventional laboratory work environment. PMID- 19560011 TI - Consensus document copyright explanation. PMID- 19560012 TI - Credentials for uterine artery embolization. PMID- 19560013 TI - Training, competency, and credentialing standards for diagnostic cervicocerebral angiography, carotid stenting, and cerebrovascular intervention: a joint statement from the American Academy of Neurology, the American Association of Neurological Surgeons, the American Society of Interventional and Therapeutic Neuroradiology, the American Society of Neuroradiology, the Congress of Neurological Surgeons, the AANS/CNS Cerebrovascular Section, and the Society of Interventional Radiology. PMID- 19560014 TI - Carotid stenting, stroke prevention, and training. PMID- 19560015 TI - Patient care and uterine artery embolization for leiomyomata. PMID- 19560016 TI - Intracranial angioplasty and stenting for cerebral atherosclerosis: a position statement of the American Society of Interventional and Therapeutic Neuroradiology, Society of Interventional Radiology, and the American Society of Neuroradiology. PMID- 19560017 TI - Society of Interventional Radiology position statement on chemoembolization of hepatic malignancies. PMID- 19560018 TI - Simulation devices in interventional radiology: validation pending. PMID- 19560019 TI - Position statement on percutaneous vertebral augmentation: a consensus statement developed by the American Society of Interventional and Therapeutic Neuroradiology, Society of Interventional Radiology, American Association of Neurological Surgeons/Congress of Neurological Surgeons, and American Society of Spine Radiology. PMID- 19560020 TI - Society of Interventional Radiology position statement: treatment of acute iliofemoral deep vein thrombosis with use of adjunctive catheter-directed intrathrombus thrombolysis. PMID- 19560021 TI - The development process for the clinical associate series. PMID- 19560022 TI - Position statement: the role of physician assistants in interventional radiology. PMID- 19560023 TI - Society of Interventional Radiology position statement on percutaneous radiofrequency ablation for the treatment of liver tumors. AB - Focal tumor ablation--whether applied percutanously, laparoscopically, or by means of open surgery--is an effective therapy for selected liver tumors. The choice of liver ablation as well as the choice between percutaneous and surgical approaches is dependent on tumor factors, patient factors, and other viable treatment options. Currently, the largest cumulative reported experience is with radiofrequency (RF) ablation of hepatocellular carcinoma and colorectal metastases. This document is a position statement of the Interventional Oncology Task Force and the Standards Division of the Society of Interventional Radiology regarding the use of percutaneous RF ablation for the treatment of liver tumors. PMID- 19560024 TI - Reporting standards for carotid artery angioplasty and stent placement. PMID- 19560025 TI - Reporting standards for inferior vena caval filter placement and patient follow up: supplement for temporary and retrievable/optional filters. PMID- 19560026 TI - Image-guided tumor ablation: standardization of terminology and reporting criteria. AB - The field of interventional oncology with use of image-guided tumor ablation requires standardization of terminology and reporting criteria to facilitate effective communication of ideas and appropriate comparison between treatments that use different technologies, such as chemical (ethanol or acetic acid) ablation, and thermal therapies, such as radiofrequency (RF), laser, microwave, ultrasound, and cryoablation. This document provides a framework that will hopefully facilitate the clearest communication between investigators and will provide the greatest flexibility in comparison between the many new, exciting, and emerging technologies. An appropriate vehicle for reporting the various aspects of image-guided ablation therapy, including classification of therapies and procedure terms, appropriate descriptors of imaging guidance, and terminology to define imaging and pathologic findings, are outlined. Methods for standardizing the reporting of follow-up findings and complications and other important aspects that require attention when reporting clinical results are addressed. It is the group's intention that adherence to the recommendations will facilitate achievement of the group's main objective: improved precision and communication in this field that lead to more accurate comparison of technologies and results and, ultimately, to improved patient outcomes. The intent of this standardization of terminology is to provide an appropriate vehicle for reporting the various aspects of image-guided ablation therapy. PMID- 19560027 TI - Reporting standards for endovascular treatment of lower extremity deep vein thrombosis. PMID- 19560028 TI - Reporting standards for percutaneous thermal ablation of renal cell carcinoma. PMID- 19560030 TI - Transcatheter therapy for hepatic malignancy: standardization of terminology and reporting criteria. AB - The field of interventional oncology includes tumor ablation as well as the use of transcatheter therapies such as embolization, chemoembolization, and radioembolization. Terminology and reporting standards for tumor ablation have been developed. The development of standardization of terminology and reporting criteria for transcatheter therapies should provide a similar framework to facilitate the clearest communication among investigators and provide the greatest flexibility in comparing established and emerging technologies. An appropriate vehicle for reporting the various aspects of catheter directed therapy is outlined, including classification of therapies and procedure terms, appropriate descriptors of imaging guidance, and terminology to define imaging and pathologic findings. Methods for standardizing the reporting of outcomes toxicities, complications, and other important aspects that require attention when reporting clinical results are addressed. It is the intention of the group that adherence to the recommendations will facilitate achievement of the group's main objective: improved precision and communication for reporting the various aspects of transcatheter management of hepatic malignancy that will translate to more accurate comparison of technologies and results and, ultimately, to improved patient outcomes. PMID- 19560029 TI - Recommended reporting standards for endovenous ablation for the treatment of venous insufficiency: joint statement of the American Venous Forum and the Society of Interventional Radiology. PMID- 19560031 TI - Reporting standards for endovascular repair of saccular intracranial cerebral aneurysms. AB - BACKGROUND AND PURPOSE: The goal of this article is to provide consensus recommendations for reporting standards, terminology, and written definitions when reporting on the radiological evaluation and endovascular treatment of intracranial, cerebral aneurysms. These criteria can be used to design clinical trials, to provide uniformity of definitions for appropriate selection and stratification of patients, and to allow analysis and meta-analysis of reported data. METHODS: This article was written under the auspices of the Joint Writing Group of the Technology Assessment Committee, Society of NeuroInterventional Surgery, Society of Interventional Radiology; Joint Section on Cerebrovascular Neurosurgery of the American Association of Neurological Surgeons and Congress of Neurological Surgeons; and Section of Stroke and Interventional Neurology of the American Academy of Neurology. A computerized search of the National Library of Medicine database of literature (PubMed) from January 1991 to December 2007 was conducted with the goal to identify published endovascular cerebrovascular interventional data about the assessment and endovascular treatment of cerebral aneurysms useful as benchmarks for quality assessment. We sought to identify those risk adjustment variables that affect the likelihood of success and complications. This article offers the rationale for different clinical and technical considerations that may be important during the design of clinical trials for endovascular treatment of cerebral aneurysms. Included in this guidance article are suggestions for uniform reporting standards for such trials. These definitions and standards are primarily intended for research purposes; however, they should also be helpful in clinical practice and applicable to all publications. CONCLUSIONS: The evaluation and treatment of brain aneurysms often involve multiple medical specialties. Recent reviews by the American Heart Association have surveyed the medical literature to develop guidelines for the clinical management of ruptured and unruptured cerebral aneurysms. Despite efforts to synthesize existing knowledge on cerebral aneurysm evaluation and treatment, significant inconsistencies remain in nomenclature and definition for research and reporting purposes. These operational definitions were selected by consensus of a multidisciplinary writing group to provide consistency for reporting on imaging in clinical trials and observational studies involving cerebral aneurysms. These definitions should help different groups to publish results that are directly comparable. PMID- 19560032 TI - Reporting standards for angioplasty and stent-assisted angioplasty for intracranial atherosclerosis. AB - BACKGROUND AND PURPOSE: Intracranial cerebral atherosclerosis causes ischemic stroke in a significant number of patients. Technological advances over the past 10 years have enabled endovascular treatment of intracranial atherosclerotic stenosis. The number of patients treated with angioplasty or stent-assisted angioplasty for this condition is increasing. Given the lack of universally accepted definitions, the goal of this document is to provide consensus recommendations for reporting standards, terminology, and written definitions when reporting clinical and radiological evaluation, technique, and outcome of endovascular treatment using angioplasty or stent-assisted angioplasty for stenotic and occlusive intracranial atherosclerosis. SUMMARY OF REPORT: This article was written under the auspices of Joint Writing Group of the Technology Assessment Committee, Society of NeuroInterventional Surgery, Society of Interventional Radiology; Joint Section on Cerebrovascular Neurosurgery of the American Association of Neurological Surgeons and Congress of Neurological Surgeons; and the Section of Stroke and Interventional Neurology of the American Academy of Neurology. A computerized search of the National Library of Medicine database of literature (PubMed) from January 1997 to December 2007 was conducted with the goal to identify published endovascular cerebrovascular interventional data in stenotic intracranial atherosclerosis that could be used as benchmarks for quality assessment. We sought to identify those risk adjustment variables that affect the likelihood of success and complications. This document offers the rationale for different clinical and technical considerations that may be important during the design of clinical trials for endovascular treatment of intracranial stenotic and occlusive atherosclerosis. Included in this guidance document are suggestions for uniform reporting standards for such trials. These definitions and standards are primarily intended for research purposes; however, they should also be helpful in clinical practice and applicable to all publications. CONCLUSION: In summary, the definitions proposed represent recommendations for constructing useful research data sets. The intent is to facilitate production of scientifically rigorous results capable of reliable comparisons between and among similar studies. In some cases, the definitions contained here are recommended by consensus of a panel of experts in this writing group for consistency in reporting and publication. These definitions should allow different groups to publish results that are directly comparable. PMID- 19560033 TI - Emerging technologies articles. PMID- 19560035 TI - Exploring the human genome in cancer with genomic approaches. PMID- 19560034 TI - High-throughput biology in the postgenomic era. PMID- 19560036 TI - Molecular imaging: a primer for interventionalists and imagers. AB - The characterization of human diseases by their underlying molecular and genomic aberrations has been the hallmark of molecular medicine. From this, molecular imaging has emerged as a potentially revolutionary discipline that aims to visually characterize normal and pathologic processes at the cellular and molecular levels within the milieu of living organisms. Molecular imaging holds promise to provide earlier and more precise disease diagnosis, improved disease characterization, and timely assessment of therapeutic response. This primer is intended to provide a broad overview of molecular imaging with specific focus on future clinical applications relevant to interventional radiology. PMID- 19560037 TI - Three-dimensional C-arm cone-beam CT: applications in the interventional suite. AB - C-arm cone-beam computed tomography (CT) with a flat-panel detector represents the next generation of imaging technology available in the interventional radiology suite and is predicted to be the platform for many of the three dimensional (3D) roadmapping and navigational tools that will emerge in parallel with its integration. The combination of current and unappreciated capabilities may be the foundation on which improvements in both safety and effectiveness of complex vascular and nonvascular interventional procedures become possible. These improvements include multiplanar soft tissue imaging, enhanced pretreatment target lesion roadmapping and guidance, and the ability for immediate multiplanar posttreatment assessment. These key features alone may translate to a reduction in the use of iodinated contrast media, a decrease in the radiation dose to the patient and operator, and an increase in the therapeutic index (increase in safety-vs-benefit ratio). In routine practice, imaging information obtained with C-arm cone-beam CT provides a subjective level of confidence factor to the operator that has not yet been thoroughly quantified. PMID- 19560038 TI - C-arm cone-beam CT: general principles and technical considerations for use in interventional radiology. AB - Digital flat-panel detector cone-beam computed tomography (CBCT) has recently been adapted for use with C-arm systems. This configuration provides projection radiography, fluoroscopy, digital subtraction angiography, and volumetric computed tomography (CT) capabilities in a single patient setup, within the interventional suite. Such capabilities allow the interventionalist to perform intraprocedural volumetric imaging without the need for patient transportation. Proper use of this new technology requires an understanding of both its capabilities and limitations. This article provides an overview of C-arm CBCT with particular attention to trade-offs between C-arm CBCT systems and conventional multi-detector CT. PMID- 19560039 TI - Emerging technologies subcommittee. PMID- 19560040 TI - Conflicts of interest in the development of new interventional medical devices. PMID- 19560041 TI - Seminars in Cell & Developmental Biology. The molecular and cellular mechanisms in the development of addiction. Editorial. PMID- 19560042 TI - Genes and signaling events that establish regional patterning of the mammalian forebrain. AB - Embryonic development of the mammalian forebrain is guided by signals from four patterning centers. The concerted actions of these signals transform the anterior neural plate and prosencephalon into discrete forebrain structures including the telencephalon (cerebral cortex and basal ganglia) and hypothalamus. In this review, we describe the signaling, transcriptional, and regulatory events that lead to induction of the prospective telencephalon, and that instruct regional development of distinct telencephalic areas along the rostrocaudal and dorsoventral axes. PMID- 19560043 TI - Histone acetylation in drug addiction. AB - Regulation of chromatin structure through post-translational modifications of histones (e.g., acetylation) has emerged as an important mechanism to translate a variety of environmental stimuli, including drugs of abuse, into specific changes in gene expression. Since alterations in gene expression are thought to contribute to the development and maintenance of the addicted state, recent efforts are aimed at identifying how drugs of abuse alter chromatin structure and the enzymes which regulate it. This review discusses how drugs of abuse alter histone acetylation in brain reward regions, through which enzymes this occurs, and ultimately what role histone acetylation plays in addiction-related behaviors. PMID- 19560044 TI - Dopamine, cocaine and the development of cerebral cortical cytoarchitecture: a review of current concepts. AB - Exposure of the developing fetus to cocaine produces lasting adverse effects on brain structure and function. Animal models show that cocaine exerts its effects by interfering with monoamine neurotransmitter function and that dopamine is cocaine's principal monoamine target in the fetal brain. This review will examine the role of dopamine receptor signaling in the regulation of normal development of the cerebral cortex, the seat of higher cognitive functions, and discuss whether dopamine receptor signaling mechanisms are the principal mediators of cocaine's deleterious effects on the ontogeny of cerebral cortical cytoarchitecture. PMID- 19560045 TI - Dopamine receptors in the learning, memory and drug reward circuitry. AB - As primary targets of a variety of abused drugs G-protein-coupled dopamine receptors in the brain play an important role in mediating the various drug induced alterations in neural and psychological processes thought to underlie the transition from voluntary drug use to habitual and progressively compulsive drug taking. This review considers the functional involvement of the five major dopamine receptor subtypes in drug reinforcement and reward and discusses the development of addiction as a series of learning transitions from initial goal directed behaviour to pathological stimulus-response habits in which drug-seeking behaviours are automatically elicited and maintained by cues and stimuli associated with drug rewards. PMID- 19560046 TI - Trafficking of dopamine transporters in psychostimulant actions. AB - Brain dopamine (DA) plays a pivotal role in drug addiction. Since the plasma membrane DA transporter (DAT) is critical for terminating DA neurotransmission, it is important to understand how DATs are regulated and this regulation impacts drug addiction. The number of cell surface DATs is controlled by constitutive and regulated endocytic trafficking. Psychostimulants impact this trafficking. Amphetamines, DAT substrates, cause rapid up-regulation and slower down regulation of DAT whereas cocaine, a DAT inhibitor, increases surface DATs. Recent reports have begun to elucidate the molecular mechanisms of these psychostimulant effects and link changes in DAT trafficking to psychostimulant induced reward/reinforcement in animal models. PMID- 19560050 TI - Mechanisms of directional asymmetry in the zebrafish epithalamus. AB - The epithalamus of zebrafish presents the best-studied case of directional asymmetry in the vertebrate brain. Epithalamic asymmetries are coupled to visceral asymmetry and include left-sided migration of a single midline structure (the parapineal organ) and asymmetric differentiation of paired bilateral nuclei (habenulae). The mechanisms underlying the establishment of epithalamic asymmetry involve the interplay between anti-symmetry and laterality signals to guide asymmetric parapineal migration. This event triggers the amplification of habenular asymmetries and the subsequent organisation of lateralised circuits in the interpeduncular nucleus. This review will summarise our current understanding on these processes and propose a sequential modular organisation of the events controlling the development of asymmetry along the parapineal-habenular interpeduncular axis. PMID- 19560048 TI - Nicotinic modulation of synaptic transmission and plasticity in cortico-limbic circuits. AB - Nicotine is the principle addictive agent delivered via cigarette smoking. The addictive activity of nicotine is due to potent interactions with nicotinic acetylcholine receptors (nAChRs) on neurons in the reinforcement and reward circuits of the brain. Beyond its addictive actions, nicotine is thought to have positive effects on performance in working memory and short-term attention related tasks. The brain areas involved in such behaviors are part of an extensive cortico-limbic network that includes relays between prefrontal cortex (PFC) and cingulate cortex (CC), hippocampus, amygdala, ventral tegmental area (VTA) and the nucleus accumbens (nAcc). Nicotine activates a broad array of nAChRs subtypes that can be targeted to pre- as well as peri- and post-synaptic locations in these areas. Thereby, nicotine not only excites different types of neurons, but it also perturbs baseline neuronal communication, alters synaptic properties and modulates synaptic plasticity. In this review we focus on recent findings on nicotinic modulation of cortical circuits and their targets fields, which show that acute and transient activation of nicotinic receptors in cortico limbic circuits triggers a series of events that affects cognitive performance in a long lasting manner. Understanding how nicotine induces long-term changes in synapses and alters plasticity in the cortico-limbic circuits is essential to determining how these areas interact in decoding fundamental aspects of cognition and reward. PMID- 19560051 TI - Abatacept therapy and safety management. AB - OBJECTIVES: To elaborate a how-to-use abatacept material intended to help physicians in the management of patients with inflammatory diseases treated with this drug in routine practice. METHODS: 1) Selection of the relevant domains by a rheumatologists' panel; 2) Search for published evidence in each domain; 3) Elaboration of the clinical tool guide with a 3-level gradation of evidence (evidence-based medicine EBM, official recommendations and expert's opinion). The experts were 11 academic rheumatologists with a large experience in prescribing abatacept and in managing rheumatoid arthritis. They were all members of the CRI (Club Rhumatismes et Inflammation), a section of the French Rheumatology Society dedicated to the inflammatory rheumatic diseases. Each fact sheet was reviewed by two other experts; 4) Regular updating based on medical literature and postmarketing surveillance data. RESULTS: Four domains were considered relevant: abatacept contraindications, management of side effects or associated diseases appearing during abatacept treatment, management of "practical situations" such as surgery or pregnancy, physician and patient information. After the literature analysis and discussion during an experts' meeting, a consensus was reached on: a pre-treatment checklist aimed at searching abatacept contraindications; a what-to do document when facing side effects or associated diseases (autoimmune pathology, bacterial or viral infections, cardiovascular diseases, intolerance to abatacept, solid or haematological malignancy) or "practical situations" (surgery, pregnancy, vaccination, travel, drug-drug interactions); an example of standard information letter to be addressed to the attending physician (rheumatologist and general practitioner); an example of standard information letter to be addressed to the patient. CONCLUSION: Based on both an EBM approach and an expert's opinion approach, this abatacept clinical tool guide should provide assistance to all physicians attending patients treated with abatacept. For a better implementation in clinical practice, this tool guide will be available online at www.cri-net.com and regularly updated. PMID- 19560052 TI - Thoracoscopic anterior instrumentation for idiopathic scoliosis. AB - COMMENTARY ON: Lonner BS, Auerbach JD, Levin R, et al. Thoracoscopic anterior instrumented fusion for adolescent idiopathic scoliosis with emphasis on the sagittal plane. Spine J 2009;9:523-9 (this issue). PMID- 19560053 TI - Thromboembolic prophylaxis for spinal injuries: is consensus possible? AB - COMMENTARY ON: Ploumis A, Ponnappan RK, Bessey JT, et al. Thromboprophylaxis in spinal trauma surgery: consensus among spine trauma surgeons. Spine J 2009;9:530 6 (this issue). PMID- 19560049 TI - Neuroimaging of children following prenatal drug exposure. AB - Recent advances in MR-based brain imaging methods have provided unprecedented capabilities to visualize the brain. Application of these methods has allowed identification of brain structures and patterns of functional activation altered in offspring of mothers who used licit (e.g., alcohol and tobacco) and illicit (e.g., cocaine, methamphetamine, and marijuana) drugs during pregnancy. Here we review that literature, which though somewhat limited by the complexities of separating the specific effects of each drug from other confounding variables, points to sets of interconnected brain structures as being altered following prenatal exposure to drugs of abuse. In particular, dopamine-rich cortical (e.g., frontal cortex) and subcortical (e.g., basal ganglia) fetal brain structures show evidence of vulnerability to intrauterine drug exposure suggesting that during brain development drugs of abuse share a specific profile of developmental neurotoxicity. Such brain malformations may shed light on mechanisms underlying prenatal drug-induced brain injury, may serve as bio-markers of significant intrauterine drug exposure, and may additionally be predictors of subsequent neuro-developmental compromise. Wider clinical use of these research-based non invasive methods will allow for improved diagnosis and allocation of therapeutic resources for affected infants, children, and young adults. PMID- 19560047 TI - Cellular events in nicotine addiction. AB - In the 25 years since the observation that chronic exposure to nicotine could regulate the number and function of high affinity nicotine binding sites in the brain there has been a major effort to link alterations in nicotinic acetylcholine receptors (nAChRs) to nicotine-induced behaviors that drive the addiction to tobacco products. Here we review the proposed roles of various nAChR subtypes in the addiction process, with emphasis on how they are regulated by nicotine and the implications for understanding the cellular neurobiology of addiction to this drug. PMID- 19560054 TI - Spine surgeons survey on attitudes regarding osteoporosis and osteomalacia screening and treatment for fractures, fusion surgery, and pseudoarthrosis. AB - COMMENTARY ON: Dipaola CP, Bible JE, Biswas D, et al. Survey of spine surgeons on attitudes regarding osteoporosis and osteomalacia screening and treatment for fractures, fusion surgery, and pseudoarthrosis. Spine J 2009;9:537-44 (this issue). PMID- 19560055 TI - Degenerative lumbar spondylolisthesis: evolution of surgical management. AB - COMMENTARY ON: Watters III WC, Bono CM, Gilbert TJ, et al. An evidence-based clinical guideline for the diagnosis and treatment of degenerative lumbar spondylolisthesis. Spine J 2009;9:609-14 (this issue). PMID- 19560056 TI - Interbody device footprint and endplate engagement characteristics: biomechanical implications. AB - COMMENTARY ON: Buttermann GR, Beaubien BP, Freeman AL, et al. Interbody device endplate engagement effects on motion segment biomechanics. Spine J 2009;9:564 573 (this issue). PMID- 19560058 TI - Technological Innovations Impact All Aspects of AMCLC 2009. PMID- 19560059 TI - The goose that lays the golden eggs. PMID- 19560060 TI - The myriad faces of imaging today (and implications for tomorrow). PMID- 19560061 TI - www.X-rayRisk.com: an online calculator for cancer risk. PMID- 19560062 TI - P4P: pragmatic for practice. PMID- 19560063 TI - Beyond substance abuse: stress, burnout, and depression as causes of physician impairment and disruptive behavior. AB - Disruptive physician behavior may diminish productivity, lead to medical errors, and compromise patient safety. The purpose of this paper is to review how common psychological conditions such as depression, stress, and burnout may drive disruptive behavior in the workplace and result in impaired patterns of professional conduct similar to what is seen with substance abuse. Problems related to these psychological morbidities may be more effectively managed with improved understanding of the conditions and behaviors, their associated risk factors, and the barriers that exist to reporting them. Further research and educational programs are warranted to address how these conditions might affect radiology. PMID- 19560064 TI - Automating quality assurance for digital radiography. AB - The existing practice of quality assurance (QA) in medical imaging is problematic because of the subjective manner in which it is performed, the lack of community and industry-wide QA standards, a paucity of supporting technology, and an overall lack of accountability. The solution for optimizing QA lies in the creation of objective and reproducible QA metrics, whose analysis can be automated through the creation of computerized QA software algorithms. The QA data derived from these computerized programs would in turn create the infrastructure for a comprehensive QA database, which can serve as a valuable resource for QA education and training, research, decision support, and technology innovation. The ability to objectively track and analyze QA practice across the wide spectrum of imaging providers creates a mechanism for the creation and refinement of objective QA standards and "best practice" guidelines. PMID- 19560065 TI - Continuous quality improvement programs provide new opportunities to drive value innovation initiatives in hospital-based radiology practices. AB - Imaging services constitute a huge portion of the of the total dollar investment within the health care enterprise. Accordingly, this generates competition among medical specialties organized along service lines for their pieces of the pie and increased scrutiny from third-party payers and government regulators. These market and political forces create challenge and opportunity for a hospital-based radiology practice. Clearly, change that creates or builds greater value for patients also creates sustainable competitive advantage for a radiology practice. The somewhat amorphous concept of quality constitutes a significant value driver for innovation in this scenario. Quality initiatives and programs seek to define and manage this amorphous concept and provide tools for a radiology practice to create or build more value. Leadership and the early adoption of these inevitable programs by a radiology practice strengthens relationships with hospital partners and slows the attrition of imaging service lines to competitors. PMID- 19560066 TI - An evaluation of MRI safety and compatibility of a silver-impregnated antimicrobial wound dressing. AB - PURPOSE: Wound infections can slow healing, increase pain, and have negative effects on a patient's quality of life. The recent emergence of antibiotic resistant bacterial strains has led wound care specialists to revisit alternative topical agents such as silver to control wound bioburden. Aquacel Ag is an ionic silver-containing barrier dressing that is able to absorb large amounts of wound exudate. The aim of this study was to assess the magnetic resonance (MR) safety and compatibility of this dressing, according to the standard requirements of the American Society for Testing and Materials (ASTM). METHODS: Radiofrequency induced temperature changes associated with the test dressing were assessed using an ASTM phantom at 123 and 64 MHz. Whether the dressing caused any image distortion or magnetic deflection or if the electric resistance of the hydrated dressing differed significantly from that of tissue was also investigated. RESULTS: Similar radiofrequency-induced temperature changes were observed during 123 MHz (nominal 3 T) MR imaging of the phantom material alone (1.3 degrees C) and when the dressing was added (1.8 degrees C-2.0 degrees C). Similar increases in temperature were also observed at 64 MHz (1.5 T) in the phantom material alone (1.4 degrees C-1.9 degrees C) and with the dressing (1.6 degrees C-1.7 degrees C). The test dressing did not cause any discernible image distortion or magnetic deflection and had similar electric resistance to human body tissues. CONCLUSION: The wound dressing impregnated with ionic silver evaluated in this study has similar magnetic and electric characteristics to human tissues and is MR safe as defined in ASTM standard F2503-05. Therefore, the dressings can be left in place when a patient is undergoing MR imaging. PMID- 19560067 TI - Endovascular repair vs open surgical repair of abdominal aortic aneurysms: comparative utilization trends from 2001 to 2006. AB - PURPOSE: Within the past few years, endovascular aneurysm repair (EVAR) has come into use for the treatment of abdominal aortic aneurysms (AAAs). In many cases, EVAR has the potential to replace traditional open surgical repair (OSR), which is more invasive, risky, and expensive. The aim of this study was to determine to what extent EVAR is replacing OSR, whether the frequency of treatment is increasing with the advent of the less invasive approach, and which specialties are performing the procedures. MATERIALS AND METHODS: The Medicare Part B data sets for 2001 through 2006 were studied. Procedure volume and utilization rates per 100,000 Medicare beneficiaries were determined for the 7 Current Procedural Terminology, fourth edition, procedure codes that describe EVAR and the 4 codes that describe OSR for AAA. Medicare's physician specialty codes were used to ascertain the specialties of the physician providers. RESULTS: A total of 31,965 OSRs for AAA were performed in Medicare beneficiaries in 2001, dropping to 15,665 by 2006 (-51%). In contrast, EVAR was carried out in 11,028 instances in 2001, increasing to 28,937 by 2006 (+162%). The utilization rate per 100,000 for OSR dropped from 90 to 42 (a rate decrease of 48) during the study period, while the rate for EVAR increased from 31 to 77 (a rate increase of 46). The combined utilization rate per 100,000 of the two types of interventions for AAA (EVAR and OSR) decreased from 121 in 2001 to 119 in 2006. In performing EVAR, procedure volume and market share in 2006 by specialty were 1) 22,003 procedures by surgeons, a 76% share; 2) 3,287 procedures by radiologists, an 11% share; 3) 1,915 procedures by cardiologists, a 7% share; and 4) 1,732 procedures by all other physicians, a 6% share. CONCLUSIONS: Treatment for AAA seems to be an example of the responsible use of new technology by physicians. The newer, less invasive, and less risky procedure (EVAR) is replacing the older and more invasive procedure (OSR) to a considerable degree. However, the overall combined utilization rate of both types of AAA treatment has remained stable in the Medicare population. There is thus no evidence to suggest that the introduction of the newer approach has led to the overtreatment of patients. Although radiologists do have a role in EVAR, surgeons strongly predominate. PMID- 19560068 TI - MR-guided focused ultrasound: a potentially disruptive technology. AB - A disruptive technology is a technological innovation that overturns the existing dominant technologies in a market. Magnetic resonance (MR)-guided focused ultrasound (MRgFUS) is a noninvasive procedure based on the combination of real time MR anatomic guidance, MR thermometry, and high-intensity focused ultrasound. Several hundred transducer elements become convergent at a point under MR guidance, leading to heating and coagulation necrosis. Outside the focal point, there is no significant heating. There is no need to break the skin for procedures in the body or to perform a craniotomy for procedures in the brain. This lack of invasiveness is what makes MRgFUS so disruptive compared with surgery. At present, MRgFUS has been used for the ablation of uterine fibroids, breast tumors, painful bony metastases, and liver tumors. In the brain, it has been used for the ablation of glioblastomas and for functional neurosurgery. Phantom and animal studies suggest future applications for prostate cancer and acute stroke treatment. PMID- 19560069 TI - Analysis of treatment planning time among systems and planners for intensity modulated radiation therapy. AB - Radiation oncology is a technologically advanced health care specialty in which numerous innovations, such as intensity-modulated radiation therapy (IMRT), require significant manpower and resources. For 3 main disease sites (prostate, head and neck, and lung), the authors investigated IMRT planning time across the United States among commonly used treatment planning systems (TPS). Treatment planning time was investigated in different components of IMRT: data transfer, contouring, beam arrangements, optimization, dose calculation, and phantom plans. The results showed significant variability among the TPS depending on the functionality and efficiency of the TPS algorithm. This study provides a road map to quantify the manpower needed and the selection of proper tools for IMRT planning and could be a model for any health care task. PMID- 19560070 TI - Why planes crash: lessons for radiology. PMID- 19560071 TI - Leading in a crisis, part 1. PMID- 19560072 TI - Multiparametric magnetic resonance imaging of breast cancer. PMID- 19560073 TI - Percutaneous image-guided ablation of renal tumors. PMID- 19560074 TI - Laws and sausages. PMID- 19560075 TI - The Maxfield family. PMID- 19560076 TI - Re: "frequency and spectrum of errors in final radiology reports generated with automatic speech recognition technology". PMID- 19560078 TI - Re: "frequency and spectrum of errors in final radiology reports generated with automatic speech recognition technology". PMID- 19560080 TI - Re: Three-dimensional left atrial and esophagus reconstruction using cardiac C arm computed tomography with image integration into fluoroscopic views for ablation of atrial fibrillation: accuracy of a novel modality in comparison with multislice computed tomography. Heart Rhythm 2008;5:1651-1657. PMID- 19560081 TI - Reactive oxidative metabolites are associated with atrial conduction disturbance in patients with atrial fibrillation. AB - BACKGROUND: Oxidative stress is associated with atrial fibrillation (AF). However, little is known about the relationship between serum markers of oxidation and electrical activity in patients with AF. OBJECTIVE: The purpose of this study was to investigate the possible association between serum markers of reactive oxidative metabolism and atrial remodeling in paroxysmal and persistent AF. METHODS: Derivatives of reactive oxidative metabolites (DROM), an index of oxidative stress, were measured in 306 consecutive patients with AF (225 paroxysmal, 81 persistent) undergoing radiofrequency (RF) catheter ablation. Filtered P-wave duration by P-wave signal-averaged ECG and levels of high sensitivity C-reactive protein (CRP) as an inflammatory marker also were measured. RESULTS: Patients were followed up for 1.2 +/- 0.8 years. DROM levels in patients with persistent AF were significantly higher than in patients with paroxysmal AF (341.6 +/- 85.5 Carratelli [Carr] units vs 305.0 +/- 77.7 Carr units, P <.001). DROM levels showed a tighter, positive correlation with filtered P-wave duration in persistent AF patients (r = 0.56, P <.001) than in all AF patients (r = 0.13, P <.05). DROM levels also showed a weaker but significant correlation with high-sensitivity CRP in patients with AF. Kaplan-Meier analysis revealed that the highest quartile of basal DROM levels exhibited a significantly higher AF recurrence rate after RF catheter ablation in patients with paroxysmal AF (P <.01). CONCLUSION: Serum markers of oxidative stress reflect atrial conduction disturbance and predict AF recurrence after RF catheter ablation in paroxysmal AF patients. DROM could serve as a biomarker for predicting risk of AF recurrence after RF catheter ablation. PMID- 19560082 TI - Stress that causes palpitations: oxidative stress and atrial fibrillation. PMID- 19560083 TI - Vectorcardiographic determinants of cardiac memory during normal ventricular activation and continuous ventricular pacing. AB - BACKGROUND: Cardiac memory (CM) refers to persistent T-wave changes on resumption of normal conduction after a period of abnormal ventricular activation. Traditionally, to observe CM, normal ventricular activation had to be restored, limiting the exploration of this phenomenon in clinical practice. OBJECTIVE: This study sought to prove that CM can be detected during continuous aberrant activation and to establish factors affecting its magnitude using a vectorcardiographic technique. METHODS: Sixteen nonpacemaker-dependent patients (11 male, age 72 +/- 8 years, mean +/- SD) undergoing pacemaker/internal cardioverter-defibrillator implantation were paced in DDD mode with a short atrioventricular (AV) delay for 7 days to induce CM. Electrocardiograms were acquired during AAI and DDD pacing at a constant rate before and after CM induction. Dower transform-derived vectorcardiograms were reconstructed and analyzed. RESULTS: T vector during AAI pacing changed in both magnitude (baseline, 0.26 +/- 0.10 mV; Day 7, 0.39 +/- 0.13 mV, P < .01) and direction aligning with the paced QRS vector (baseline DDD QRS - AAI T angle 125 degrees +/ 36 degrees; Day 7, 39 degrees +/- 21 degrees, P < .01). During DDD pacing, there was no change in T-vector direction, but T amplitude decreased (baseline, 1.06 +/ 0.32 mV; Day 7, 0.71 +/- 0.26 mV, P < .01). CM measured as T-vector peak displacement (TPD) was identical in AAI and DDD mode (TPD 0.46 +/- .0.17 mV and 0.46 +/- 0.17 mV, respectively). Individual CM magnitude correlated with QRS/T vector amplitude ratio during DDD pacing at baseline (r = 0.90). CONCLUSION: CM can be reliably shown during continuous ventricular pacing, expanding its application to situations in which abnormal ventricular activation persists. Its magnitude is determined by the QRS/T-amplitude ratio of the ventricular paced beat. PMID- 19560084 TI - Prevention of phrenic nerve injury during epicardial ablation: comparison of methods for separating the phrenic nerve from the epicardial surface. AB - BACKGROUND: The proximity of the phrenic nerve (PN) to cardiac tissue relevant to arrhythmias may increase the risk of PN injury. Strategies for preventing PN injury in the pericardial space are limited. OBJECTIVE: The purpose of this study was to compare methods for separating the PN from the epicardial surface in order to prevent PN injury. METHODS: Eight patients referred for epicardial ablation of arrhythmias were enrolled in the study. All patients required ablation near the PN. Endocardial and epicardial access was obtained in all patients. A three dimensional mapping system was used to guide mapping and ablation. All patients underwent epicardial catheter ablation. Pacing via the ablation catheter identified the location of the PN. In order to prevent PN injury, four new strategies were tested in each patient. We sought to increase the distance between the epicardium and the PN by (1) placing a large-diameter balloon between the nerve and the myocardium, (2) introducing saline in steps of 20 ml until PN capture was lost or blood pressure dropped below 60 mmHg, (3) introducing air until PN capture was lost or blood pressure dropped below 60 mmHg, or (4) introducing a combination of saline and air until PN capture was lost or blood pressure dropped below 60 mmHg. RESULTS: At each step, epicardial pacing was performed to assess for PN stimulation. The combination of air and saline resulted in the greatest decrease of PN stimulation. Saline only failed in all cases. Air only and balloon placement were infrequently successful. CONCLUSION: Controlled and progressive inflation of air and saline together with careful monitoring of hemodynamic parameters appears to be the best strategy for preventing PN injury during epicardial ablation. Placement of a large balloon in the appropriate location can be difficult. PMID- 19560085 TI - The esophageal effects of cryoenergy during cryoablation for atrial fibrillation. AB - BACKGROUND: Cryoenergy is being increasingly used for atrial fibrillation (AF) ablation, but the thermal effect of cryoenergy on the esophagus remains undefined. OBJECTIVE: This study examines the esophageal effects of cryoenergy used during AF ablation. METHODS: Catheter ablation was performed using a cryoballoon catheter in 67 AF patients (Cryoballoon group), and a spot cryocatheter to complete irrigated radiofrequency lesion sets at segments in close proximity to the esophagus in 7 AF patients (Cryo-Focal group). A temperature probe monitored the luminal esophageal temperature (LET) in all patients; LET changes did not guide therapy. Post-procedural endoscopy was performed on 35 of 67 (52%) Cryoballoon and all Cryo-Focal patients. RESULTS: Significant LET decreases (>1 degrees C) occurred in 62 of 67 (93%) Cryoballoon patients. LET continued to decrease after termination of cryoablation before recovering to normal. Temperature decreases were more pronounced during ablation at the inferior (3.1 degrees C) than superior pulmonary veins (1.5 degrees C); the lowest observed temperature was 0 degrees C. Post-procedural endoscopy showed esophageal ulcerations in 6 of 35 (17%) patients. There were no atrial-esophageal fistulas, and all ulcers had healed on follow-up endoscopy. Patients with and without ulceration differed with respect to mean LET nadir, cumulative LET decrease, and number of LETs <30 degrees C. In the Cryo-Focal group, 6 +/- 2 spot cryolesions per patient resulted in 1.3 +/- 1 LET decreases per patient, and an absolute nadir of 32.5 degrees C. CONCLUSION: Cryoballoon ablation can cause significant LET decreases, resulting in reversible esophageal ulcerations in 17% of patients. No ulcerations occurred with adjunctive spot cryoablation at regions near the esophagus during radiofrequency ablation procedures. PMID- 19560086 TI - Echocardiographic study of the optimal atrioventricular delay at rest and during exercise in recipients of cardiac resynchronization therapy systems. AB - BACKGROUND: It is unclear whether, in recipients of cardiac resynchronization therapy (CRT) systems, the optimal AV delay should be the same, shorter, or longer during exercise than at rest. OBJECTIVE: This study sought to examine the effects of atrioventricular (AV) delay optimization at rest and during exercise in 50 recipients of CRT systems. METHODS: We measured left ventricular (LV) outflow tract velocity time integral (OT-VTI) and LV filling time (FT) echocardiographically, at rest and during exercise to 60% of the maximal predicted heart rate, with the sensed AV delay set at 40, 70, 100, 120, 150, and 200 ms. The measurements made at rest versus those made during exercise, and among the several programmed AV delays, were compared. RESULTS: The optimal AV delay based on LVOT-VTI was shorter during exercise than at rest in 37%, unchanged in 37%, and longer in 26% of patients. The optimal AV delay based on LVFT was shorter during exercise than at rest in 27%, unchanged in 23%, and longer in 50% of patients. Optimization of the AV delay during exercise increased LVFT and LVOT-VTI significantly (P < .05) compared with (1) any other arbitrarily chosen AV delay, (2) the optimal AV delay at rest, (3) an AV delay systematically shortened from rest to exercise. CONCLUSION: Optimization of the AV delay had a positive effect on echocardiographic indices of LV function. The systematic shortening of the AV delay during exercise is not recommended because, in a high proportion of patients, the optimal AV delay was longer during exercise than at rest. PMID- 19560087 TI - Transiliac ICD implantation: defibrillation vector flexibility produces consistent success. AB - BACKGROUND: The transiliac approach to implantable cardioverter-defibrillator (ICD) implantation is an alternative in patients for whom pectoral placements are contraindicated. The defibrillation vector is altered from the pectoral configuration because of pulse generator placement in one of the upper abdominal quadrants and separate single-coil, active-fixation defibrillation leads positioned in the high right atrium and right ventricular apex. OBJECTIVE: The feasibility, safety, and complications of this approach and the results of defibrillation testing (DFT) with this configuration are described. METHODS: Twenty-three patients (16 male and 7 female, mean age 65.7 +/- 13.2 years) required transiliac approach to ICD placement. The leads were inserted through the iliac vein immediately superior to the inguinal ligament. When required, a subcutaneous coil was tunneled posterior to the left ventricle from the left axilla. RESULTS: The right iliac vein entry was used in 17 patients, with placement of the pulse generator in the left upper quadrant in 16 patients. Atrial and ventricular lead pacing and sensing function were acceptable. Initial defibrillation success with a safety margin of 10 J was achieved in 15 patients. With the placement of an additional subcutaneous coil in the remaining 8 patients, defibrillation success with a safety margin of 10 J was increased to 19 patients, whereas defibrillation success with a safety margin of 5 J was achieved in all patients, although 1 patient required repeat testing 24 hours after implantation. There were no acute complications. Late complications occurred in 3 patients, comprised of atrial lead malfunction, device infection, and right ventricular defibrillation lead fracture. CONCLUSION: The iliac vein approach to ICD implantation is a safe and effective alternative technique. Flexibility in lead placement, defibrillation vectors, and careful DFT are required to produce a consistently effective system. PMID- 19560089 TI - Mechanisms of stretch-induced atrial fibrillation in the presence and the absence of adrenocholinergic stimulation: interplay between rotors and focal discharges. AB - BACKGROUND: Both atrial stretch and combined adrenocholinergic stimulation (ACS) have been shown to favor initiation and maintenance of atrial fibrillation (AF). Their respective contributions to the electrophysiological mechanism remains, however, incompletely understood. OBJECTIVE: This study endeavored to determine the mechanism of maintenance of stretch-related AF (SRAF) in the presence and absence of ACS and to assess how focal discharges interact with rotors to modify the level of complexity in the activation patterns to perpetuate AF. METHODS: Video imaging of AF dynamics was carried out using a SRAF model in isolated sheep hearts (n = 24). Pharmacological approaches were used to (1) mimic ACS with acetylcholine (1 microM) plus isoproterenol (0.03 microM), and (2) abolish triggered activity, in response to sarcoplasmic reticulum calcium release, with caffeine (5 mM, CA) or ryanodine (10 to 40 microM, RYA). RESULTS: In the absence of ACS, on perfusion of CA or RYA, focal discharges were abolished and SRAF was terminated in most of the cases (10 of 13 experiments). In the presence of ACS, multiple drifting rotors as well as a large number of focal discharges were identified and only 1 of 11 AF episodes was terminated. CONCLUSIONS: In the absence of ACS, SRAF is maintained by high-frequency focal discharges that generate fibrillatory conduction and wave breaks. In the presence of ACS, SRAF dynamics is characterized by multiple high frequency rotors that are rendered unstable by spatially distributed focal discharges. PMID- 19560088 TI - Arrhythmogenic right ventricular cardiomyopathy/dysplasia clinical presentation and diagnostic evaluation: results from the North American Multidisciplinary Study. AB - BACKGROUND: Prior reports on patients with arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVC/D) focused on individuals with advanced forms of the disease. Data on the diagnostic performance of various testing modalities in newly identified individuals suspected of having ARVC/D are limited. OBJECTIVE: The purpose of the Multidisciplinary Study of Arrhythmogenic Right Ventricular Cardiomyopathy/Dysplasia was to study the clinical characteristics and diagnostic evaluation of a large group of patients newly identified with ARVC/D. METHODS: A total of 108 newly diagnosed patients with suspected ARVC/D were prospectively enrolled in the United States and Canada. The patients underwent noninvasive and invasive tests using standardized protocols that initially were interpreted by the enrolling center and adjudicated by blind analysis in six core laboratories. Patients were followed for a mean of 27 +/- 16 months (range 0.2-63 months). RESULTS: The clinical profile of these newly diagnosed patients differs from the profile of reported patients with more advanced disease. There was considerable difference in the initial and final classification of the presence of ARVC/D after the diagnostic tests were evaluated by the core laboratories. Final clinical diagnosis was 73 affected, 28 borderline, and 7 unaffected. Individual tests agreed with the final diagnosis in 50% to 70% of the 73 patients with a final classification of affected. CONCLUSION: The clinical profile of 108 newly diagnosed probands with suspected ARVC/D indicates that a combination of diagnostic tests is needed to evaluate the presence of right ventricular structural, functional, and electrical abnormalities. Echocardiography, right ventricular angiography, signal-averaged ECG, and Holter monitoring provide optimal clinical evaluation of patients suspected of ARVC/D. PMID- 19560091 TI - There is no transmural heterogeneity in an index of action potential duration in the canine left ventricle. AB - BACKGROUND: Transmural heterogeneity in ventricular repolarization demonstrated in vitro has been difficult to confirm in vivo. Whether this discrepancy reflects a physiological phenomenon or a methodological problem remains a vivid matter of debate despite a plethora of experimental work. Therefore, we have measured the relevant electrophysiological parameters first in vivo and repeated these in the same heart and at identical sites in vitro. Methodological issues were tackled by using both unipolar and bipolar recordings. Physiological issues were explored by measuring both local and functional electrophysiological parameters. METHODS: In 10 healthy dogs, 2 high-resolution needle electrodes were inserted into the left ventricle. Effective refractory periods (ERP) as well as activation recovery intervals (ARI) were determined at each electrode along both needles at basic cycle lengths (BCL) of 850 and 300 ms, respectively. After excision of the heart, ERP and ARI measurements were repeated in the arterially perfused wedge preparations. RESULTS: First, we observed that ERPs and ARIs were significantly shorter in vivo than in vitro. Mean ERPs and ARIs of all muscle layers were relatively uniform throughout the ventricular wall in vivo. The transition from the in vivo to the in vitro preparation was associated with a significant albeit small increase of mean ARIs in the subendocardium, whereas interlayer differences in mean ERPs did not reach statistical significance as in vivo. CONCLUSION: In the intact canine left ventricular wall, a more or less homogeneous distribution in transmural ERP and ARI is present. PMID- 19560092 TI - Cardiac resynchronization therapy and the electrical substrate in heart failure: what does the QRS conceal? PMID- 19560090 TI - Multiple monophasic shocks improve electrotherapy of ventricular tachycardia in a rabbit model of chronic infarction. AB - BACKGROUND: We previously showed that the cardioversion threshold (CVT) for ventricular tachycardia (VT) is phase dependent when a single monophasic shock (1MP) is used. OBJECTIVE: The purpose of this study was to extend these findings to a biphasic shock (1BP) and to compare the efficacy of phase-independent multiple monophasic (5MP) and biphasic shocks (5BP). METHODS: Panoramic optical mapping with blebbistatin (5 microM) was performed in postmyocardial infarction rabbit hearts (n = 8). Flecainide (1.64 +/- 0.68 microM) was administered to promote sustained arrhythmias. 5MP and 5BP were applied within one VT cycle length (CL). Results were compared to 1BP and antitachycardia pacing. RESULTS: We observed monomorphic VT with CL = 149.6 +/- 18.0 ms. Similar to 1MP, CVTs of 1BP were found to be phase dependent, and the maximum versus minimum CVT was 8.6 +/- 1.7 V/cm versus 3.7 +/- 1.9 V/cm, respectively (P = .0013). Efficacy of 5MP was higher than that of 1BP and 5BP. CVT was 3.2 +/- 1.4 V/cm for 5MP versus 5.3 +/- 1.9 V/cm for 5BP (P = .00027). 5MP versus averaged 1BP CVT was 3.6 +/- 2.1 V/cm versus. 6.8 +/- 1.5 V/cm, respectively (P = .00024). Antitachycardia pacing was found to be completely ineffective in this model. CONCLUSION: Maintenance of shock-induced virtual electrode polarization by multiple monophasic shocks over a VT cycle is responsible for unpinning of reentry leading to self-termination. Elimination of virtual electrode polarization by shock polarity reversal during multiple biphasic shocks proved ineffective. A significant reduction in CVT can be achieved by applying multiple monophasic shocks within one VT CL or one single shock at the proper coupling interval. PMID- 19560093 TI - Inappropriate shock from nonphysiologic noise during implantation of a Boston Scientific COGNIS N119 biventricular implantable cardioverter-defibrillator (CRT D). PMID- 19560094 TI - Catheter ablation in tetralogy of Fallot. PMID- 19560095 TI - Cardiac electrophysiology in Bolivia. PMID- 19560096 TI - Remote monitoring of implantable devices: the European experience. PMID- 19560098 TI - Transvenous lead extraction: Heart Rhythm Society expert consensus on facilities, training, indications, and patient management: this document was endorsed by the American Heart Association (AHA). PMID- 19560100 TI - Development and validation of the Memory Performance Index: reducing measurement error in recall tests. AB - BACKGROUND: The Memory Performance Index (MPI) quantifies the pattern of recalled and nonrecalled words of the Consortium to Establish a Registry for Alzheimer's Disease Wordlist (CWL) onto a 0 to 100 scale and distinguishes normal from mild cognitive impairment with 96% to 97% accuracy. METHODS: In group A, 121,481 independently living individuals, 18 to 106 years old, were assessed with the CWL and classified as cognitively impaired (N = 5,971) or normal (N = 115,510). The MPI and CWL immediate free recall (IFR), delayed free recall (DFR), and total free recall (TFR) scores (the outcome measures) were each regressed against predictors of age, gender, race, education, test administration method (in-person or telephone), and wordlist used. Predictor effect sizes (Cohen's f(2)) were computed for each outcome. In addition, CWL plus Functional Assessment Staging Tests (FAST) were administered to 441 normal to moderately severely demented (FAST stages 1 to 6) patients (group B). Median MPI scores were tested for significant differences across FAST stage. RESULTS: For group A, the variance explained by all predictors combined was MPI = 55.0%, IFR = 24.9%, DFR = 23.4%, and TFR = 26.9%. The age effect size on MPI score was large, but it was small on IFR, DFR, and TFR. The other predictors all had negligible (<0.02) or small effect sizes (0.02 to 0.15). For group B, median MPI scores progressively declined across all FAST stages (P < .0002). CONCLUSIONS: MPI score progressively declines with increasing dementia severity. Also, MPI score explains 2 to 3 times more variance than total scores, which improves ability to detect treatment effects. PMID- 19560099 TI - Exercise improves cognition and hippocampal plasticity in APOE epsilon4 mice. AB - BACKGROUND: Human studies on exercise, cognition, and apolipoprotein E (APOE) genotype show that epsilon4 carriers may benefit from regular physical activity. METHODS: We examined voluntary wheel-running, memory, and hippocampal plasticity in APOE epsilon3 and APOE epsilon4 transgenic mice at 10-12 months of age. RESULTS: Sedentary epsilon4 mice exhibited deficits in cognition on the radial arm water maze (RAWM), a task dependent on the hippocampus. Six weeks of wheel running in epsilon4 mice resulted in improvements on the RAWM to the level of epsilon3 mice. Hippocampal brain-derived neurotrophic factor (BDNF) levels were similar in epsilon3 and epsilon4 mice, and after exercise BDNF was similarly increased in both epsilon3 and epsilon4 mice. In sedentary epsilon4 mice, tyrosine kinase B (Trk B) receptors were reduced by 50%. Exercise restored Trk B in epsilon4 mice to the level of epsilon3 mice, and in epsilon4 mice, exercise dramatically increased synaptophysin, a marker of synaptic function. CONCLUSIONS: Our results support the hypothesis that exercise can improve cognitive function, particularly in epsilon4 carriers. PMID- 19560101 TI - Markers of Alzheimer's disease in a population attending a memory clinic. AB - BACKGROUND: New marker-based criteria for the diagnosis of Alzheimer's disease (AD) were recently proposed. We describe their operational translation in 144 consecutive patients referred to our Memory Clinic. METHODS: Visual ratings of hippocampal atrophy and of cortical glucose hypometabolism in magnetic resonance imaging and positron emission tomography, and concentrations of total tau and Abeta1-42 in cerebrospinal fluid were assessed in 12 patients with subjective memory complaints (SMCs) (Mini-Mental State Examination [MMSE] score, 28.0 +/- 1.1 [mean +/- SD]), 37 with mild cognitive impairment (MCI) (MMSE, 25.1 +/- 3.6), 55 with AD (MMSE, 21.1 +/- 3.5), and 40 with non-AD dementia (MMSE, 21.6 +/- 5.5). RESULTS: The sensitivity for AD of each individual biomarker was higher (65% to 87%) than for MCI (18% to 50%). Each biomarker's specificity for SMC and non-AD dementias was good to moderate (83% and 53%). Positivity for at least one marker increased the probability 38 times of belonging to the AD group (P < 0.0001). CONCLUSION: The new diagnostic criteria can be operationalized in clinical routines, but longitudinal studies of MCI patients will need to assess the criteria's prognostic value. PMID- 19560102 TI - Elevated C-reactive protein levels are associated with prevalent dementia in the oldest-old. AB - BACKGROUND: C-reactive protein (CRP) is a nonspecific marker of inflammation that is increased in the brain and serum of patients with Alzheimer's disease (AD), and has been associated with increased risk of developing dementia. Inflammation increases with age, and the number of people reaching age 90 years and older is growing, making the association between inflammation and dementia increasingly relevant. Using a cross-sectional design, we examined whether high levels of serum CRP are associated with increased odds of prevalent dementia in the oldest old. METHODS: Serum CRP levels of 305 participants (mean age +/- standard deviation, 94.3 +/- 2.9 years) from the 90+ Study, a longitudinal cohort study of people aged 90 years and older, were evaluated with respect to all-cause dementia. Levels of CRP were divided into three categories: undetectable (<0.5 mg/dL), detectable (0.5-0.7 mg/dL), and elevated (> or =0.8 mg/dL). Odds ratios (ORs) were calculated using logistic regression, and were adjusted for covariates. RESULTS: Relative to participants with undetectable CRP levels, participants with detectable or elevated CRP levels had increased odds of all cause dementia (detectable: OR, 3.0; 95% confidence interval, 1.2-7.3; elevated: OR, 5.0; 95% confidence interval, 1.9-12.9). When participants were subdivided by gender, significantly increased ORs were seen only in women. CONCLUSIONS: In the oldest-old, high CRP levels are associated with increased odds of all-cause dementia, particularly in women. Prospective studies are necessary to confirm whether increased CRP levels are associated with an increased risk of developing dementia in this age group. PMID- 19560103 TI - Scales as outcome measures for Alzheimer's disease. AB - The assessment of patient outcomes in clinical trials of new therapeutics for Alzheimer's disease (AD) continues to evolve. In addition to assessing drugs for symptomatic relief, an increasing number of trials are focusing on potential disease-modifying agents. Moreover, participants with AD are being studied earlier in their course of disease. As a result, the limitations of current outcome measures have become more apparent, as has the need for better instruments. In recognition of the need to review and possibly revise current assessment measures, the Alzheimer's Association, in cooperation with industry leaders and academic investigators, convened a Research Roundtable meeting devoted to scales as outcome measures for AD clinical trials. The meeting included a discussion of methodological issues in the use of scales in AD clinical trials, including cross-cultural issues. Specific topics related to the use of cognitive, functional, global, and neuropsychiatric scales were also presented. Speakers also addressed academic and industry initiatives for pooling data from untreated and placebo-treated patients in clinical trials. A number of regulatory topics were also discussed with agency representatives. Panel discussions highlighted areas of controversy, in an effort to gain consensus on various topics. PMID- 19560105 TI - Alzheimer's disease and infection: do infectious agents contribute to progression of Alzheimer's disease? AB - Infection with several important pathogens could constitute risk factors for cognitive impairment, dementia, and Alzheimer's disease (AD) in particular. This review summarizes the data related to infectious agents that appear to have a relationship with AD. Infections with herpes simplex virus type 1, picornavirus, Borna disease virus, Chlamydia pneumoniae, Helicobacter pylori, and spirochete were reported to contribute to the pathophysiology of AD or to cognitive changes. Based on these reports, it may be hypothesized that central nervous system or systemic infections may contribute to the pathogenesis or pathophysiology of AD, and chronic infection with several pathogens should be considered a risk factor for sporadic AD. If this hypothesis holds true, early intervention against infection may delay or even prevent the future development of AD. PMID- 19560104 TI - Amyloid precursor protein transgenic mouse models and Alzheimer's disease: understanding the paradigms, limitations, and contributions. AB - Transgenic (Tg) mice that overexpress mutant familial Alzheimer's disease (AD) amyloid precursor protein (APP) genes have contributed to an understanding of dementia pathology, and support the amyloid cascade hypothesis. Although many sophisticated mice APP models exist, none recapitulates AD cellular and behavioral pathology. The morphological resemblance to AD amyloidosis is impressive, but fundamental biophysical and biochemical properties of the APP/Abeta produced in Tg mice differ substantially from those of humans. The greater resilience of Tg mice in the presence of substantial Abeta burdens suggests that levels and forms deleterious to human neurons are not as noxious in these models. Transgenic mice were widely used for testing AD therapeutic agents, and demonstrated promising results. Unfortunately, clinical trials resulted in unforeseen adverse events or negative therapeutic outcomes. The disparity between success and failure is in part attributable to evolutionary divergence between humans and rodents. These observations suggest that the pathogenesis of AD is by far more intricate than can be explained by a straightforward accumulation of Abeta. PMID- 19560107 TI - A feature-based inference model of numerical estimation: the split-seed effect. AB - Prior research has identified two modes of quantitative estimation: numerical retrieval and ordinal conversion. In this paper we introduce a third mode, which operates by a feature-based inference process. In contrast to prior research, the results of three experiments demonstrate that people estimate automobile prices by combining metric information associated with two critical features: product class and brand status. In addition, Experiments 2 and 3 demonstrated that when participants are seeded with the actual current base price of one of the to-be estimated vehicles, they respond by revising the general metric and splitting the information carried by the seed between the two critical features. As a result, the degree of post-seeding revision is directly related to the number of these features that the seed and the transfer items have in common. The paper concludes with a general discussion of the practical and theoretical implications of our findings. PMID- 19560106 TI - Creating a transatlantic research enterprise for preventing Alzheimer's disease. AB - In recognition of the global problem posed by Alzheimer's disease and other dementias, an international think-tank meeting was convened by Biocat, the Pasqual Maragall Foundation, and the Lou Ruvo Brain Institute in February 2009. The meeting initiated the planning of a European Union-North American collaborative research enterprise to expedite the delay and ultimate prevention of dementing disorders. The key aim is to build parallel and complementary research infrastructure that will support international standardization and inter operability among researchers in both continents. The meeting identified major challenges, opportunities for research resources and support, integration with ongoing efforts, and identification of key domains to influence the design and administration of the enterprise. PMID- 19560110 TI - Auricular acupuncture as a treatment for pregnant women who have low back and posterior pelvic pain: a pilot study. AB - OBJECTIVE: The primary aim of this study was to examine whether 1 week of continuous auricular acupuncture could reduce low back and posterior pelvic pain associated with pregnancy. STUDY DESIGN: A randomized controlled trial was conducted on pregnant women who have lower back and posterior pelvic pain. These women were randomly assigned into an acupuncture group, a sham acupuncture group, or a waiting list control group. All participants were monitored for 2 weeks. RESULTS: Baseline and day 7 showed significant group differences in pain (F = 15; P < .0001) and in the disability rating index score (F = 7; P < .0001). The participants in the acupuncture group reported a significant reduction of pain and improvement of functional status as compared with those in the sham acupuncture and control groups. CONCLUSION: One week of continuous auricular acupuncture decreases the pain and disability experienced by women with pregnancy related low back and posterior pelvic pain. PMID- 19560108 TI - Differential regulation of genes for fetoplacental lipid pathways in pregnancy with gestational and type 1 diabetes mellitus. AB - OBJECTIVE: Changes in metabolic homeostasis in pregnant diabetic women are potential determinants of increased adiposity of the fetus. The aim of this study was to characterize diabetes mellitus-induced changes in genes for fetoplacental energy metabolism in relation to fetal adiposity. STUDY DESIGN: Placentas of women with type 1 diabetes mellitus, gestational diabetes mellitus (GDM), or no complications were analyzed by microarray profiling. The pattern of gene expression was assessed in primary placental cell cultures. RESULTS: Diabetes mellitus was associated with 49 alterations in gene expression at key steps in placental energy metabolism, with 67% of the alterations related to lipid pathways and 9% of the alterations related to glucose pathways. Preferential activation of lipid genes was observed in pregnancy with GDM. Type 1 diabetes mellitus induced fewer lipid modifications but an enhancement of glycosylation and acylation pathways. Oleate enhanced expression of genes for fatty acid esterification and the formation of lipid droplets 3 times as much as glucose in cultured placental cells. CONCLUSION: These results point to fatty acids as preferential lipogenic substrates for placental cells and suggest that genes for fetoplacental lipid metabolism are enhanced selectively in GDM. The recruited genes may be instrumental in increasing transplacental lipid fluxes and the delivery of lipid substrates for fetal use. PMID- 19560111 TI - Comparison between lidocaine-prilocaine cream (EMLA) and mepivacaine infiltration for pain relief during perineal repair after childbirth: a randomized trial. AB - OBJECTIVE: The purpose of this study was to compare the effectiveness of topically applied lidocaine-prilocaine (EMLA) cream with local anesthetic infiltration in the reduction of pain during perineal suturing after childbirth. STUDY DESIGN: Sixty-one women with either an episiotomy or a perineal laceration after vaginal delivery were assigned randomly to receive either the application of EMLA cream (n = 31) or infiltration with mepivacaine (n = 30) before perineal suturing. Primary outcome was pain during perineal repair. RESULTS: Women in the EMLA group had lower pain scores than those in the mepivacaine group (1.7 +/- 2.4 vs 3.9 +/- 2.4; P = .0002). The proportion of women who needed additional anesthesia was similar in the 2 groups (3/30 vs 5/31; P = .71). A significantly higher proportion of women expressed satisfaction with anesthesia method in the EMLA group, compared with the mepivacaine group (83.8% vs 53.3%; P = .01) CONCLUSION: EMLA cream appears to be an effective and satisfactory alternative to local anesthetic infiltration for the relief of pain during perineal repair. PMID- 19560112 TI - Interval to spontaneous delivery after elective removal of cerclage. AB - OBJECTIVE: The purpose of this study was to estimate the time interval between elective cerclage removal and spontaneous delivery. METHODS: Singleton pregnancies with McDonald cerclage were evaluated for the interval between elective cerclage removal (36-37 weeks) and spontaneous delivery. We also compared spontaneous delivery within 48 hours after cerclage removal between women with ultrasound-indicated vs history-indicated cerclage. RESULTS: We identified 141 women with elective cerclage removal. The mean interval between removal and delivery was 14 days. Only 11% of women delivered within 48 hours. Women with ultrasound-indicated cerclage were more likely to deliver within 48 hours, compared with women with history-indicated cerclage (odds ratio, 5.14; 95% confidence interval, 1.10-24.05). CONCLUSION: The mean interval between elective cerclage removal and spontaneous delivery is 14 days. Women with cerclage who achieved 36-37 weeks should be counseled that their chance of spontaneous delivery within 48 hours after elective cerclage removal is only 11%. PMID- 19560116 TI - Adjustment to termination of pregnancy for fetal anomaly: a longitudinal study in women at 4, 8, and 16 months. AB - OBJECTIVE: We studied psychological outcomes and predictors for adverse outcome in 147 women 4, 8, and 16 months after termination of pregnancy for fetal anomaly. STUDY DESIGN: We conducted a longitudinal study with validated self completed questionnaires. RESULTS: Four months after termination 46% of women showed pathological levels of posttraumatic stress symptoms, decreasing to 20.5% after 16 months. As to depression, these figures were 28% and 13%, respectively. Late onset of problematic adaptation did not occur frequently. Outcome at 4 months was the most important predictor of persistent impaired psychological outcome. Other predictors were low self-efficacy, high level of doubt during decision making, lack of partner support, being religious, and advanced gestational age. Strong feelings of regret for the decision were mentioned by 2.7% of women. CONCLUSION: Termination of pregnancy for fetal anomaly has significant psychological consequences for 20% of women up to > 1 year. Only few women mention feelings of regret. PMID- 19560115 TI - Risk factors for failure of retropubic and transobturator midurethral slings. AB - OBJECTIVE: The aim of our study was to identify and compare risk factors for failure of retropubic and transobturator procedures. STUDY DESIGN: This was a retrospective cohort study. Women with predominant stress urinary incontinence who underwent a retropubic (n = 214) or transobturator tape procedure (n = 173) were included. Therapy was considered to have failed in women reporting any amount of urine leakage during stress after 2 and/or 12 months. RESULTS: Risk factors for failure were mixed urinary incontinence (MUI; odds ratio [OR], 3.7; 95% confidence interval [CI], 1.5-9.1) and the observation of detrusor overactivity (DO) at urodynamics (OR, 8.6; 95% CI, 1.9-39.4) in the retropubic group. Reporting a history of previous incontinence surgery (OR, 3.9; 95% CI, 1.3 11.7) and a low mean urethral closure pressure (MUCP) at urodynamics (OR, 14.5; 95% CI, 1.5-139.0) were risk factors for failure in the transobturator group. CONCLUSION: Women with previous incontinence surgery or a low MUCP might benefit more from a retropubic sling, whereas a transobturator procedure might be preferable in women with MUI or DO. PMID- 19560114 TI - Childbearing is associated with higher incidence of the metabolic syndrome among women of reproductive age controlling for measurements before pregnancy: the CARDIA study. AB - OBJECTIVE: We sought to prospectively examine whether childbearing is associated with higher incidence of the metabolic syndrome (MetS) after delivery among women of reproductive age. STUDY DESIGN: In 1451 nulliparas who were aged 18-30 years and free of the MetS at baseline (1985-1986) and reexamined up to 4 times during 20 years, we ascertained incident MetS defined by the National Cholesterol Education Program Adult Treatment Panel III criteria among time-dependent interim birth groups by gestational diabetes mellitus (GDM): (0 [referent], 1 non-GDM, 2+ non-GDM, 1+ GDM births). Complementary log-log models estimated relative hazards of the MetS among birth groups adjusted for race, age, and baseline and follow-up covariates. RESULTS: We identified 259 incident MetS cases in 25,246 person-years (10.3/1000 person-years). Compared with 0 births, adjusted relative hazards (95% confidence interval [CI]) were 1.33 (95% CI, 0.93-1.90) for 1 non-GDM, 1.62 (95% CI, 1.16-2.26) for 2+ non-GDM (P trend = .02), and 2.43 (95% CI, 1.53-3.86) for 1+ GDM births. CONCLUSION: Increasing parity is associated with future development of the MetS independent of prior obesity and pregnancy-related weight gain. Risk varies by GDM status. PMID- 19560117 TI - Risk of invasive cervical cancer in relation to management of abnormal Pap smear results. AB - OBJECTIVE: We sought to evaluate the management of women with abnormal cytology in terms of subsequent risk of invasive cervical cancer. STUDY DESIGN: The screening histories of all invasive cervical cancer cases diagnosed in Sweden 1999-2001 and of 5 population-based controls per case were reviewed. In all, 159 patients and 258 control subjects aged < 67 years had an abnormal smear result 0.5-6.5 years prior to cancer diagnosis. The cervical cancer risk was estimated in relation to management by calculating odds ratios. RESULTS: Histologic assessment of low-grade squamous abnormalities strongly reduced the risk compared to repeated cytology (odds ratio, 0.46; 95% confidence interval, 0.24-0.89). Delaying histologic assessment was also associated with a higher risk (odds ratio, 5.65; 95% confidence interval, 1.39-23.05). After high-grade squamous atypia, absence of any cytologic or histologic specimen was a major determinant of cancer risk (odds ratio, 12.52; 95% confidence interval, 1.42-infinitive). CONCLUSION: For adequate protection against invasive cervical cancer, further assessment with histology must be recommended also for women with low-grade squamous abnormalities. PMID- 19560119 TI - The more he looked inside, the more piglet wasn't there: is autism really blessed with visual hyperacuity? PMID- 19560118 TI - Periodontal histomorphometry and status of aged sheep subjected to ovariectomy, malnutrition and glucocorticoid application. AB - OBJECTIVE: Ageing, hypogonadism, malnutrition, and the application of glucocorticoids have adverse effects on skeletal homeostasis. Herein we determined to which extent the periodontium undergoes catabolic changes under these conditions in a sheep model. METHODS: Six old sheep with a mean age of 7.5+/-1.0 years were subjected to ovariectomy, calcium/vitamin D-restricted diet, and intramuscular administration of approximately 2g methylprednisolone. Six adult sheep with a mean age of 3.8+/-0.9 years remained untreated and served as controls. First and second premolars of both jaws were subjected to histological analysis. The distances from the gingival margin (GM) and from the alveolar bone crest (ABC) to the cemento-enamel junction (CEJ) were determined. Periodontal attachment was given as the ratio between the dimension of the periodontal ligament and the alveolar bone. Clinical data were collected by counting the number of teeth missing, teeth with gingival recession, and teeth with a probing depth > 4 mm. RESULTS: We report that distance between GM and CEJ (2.1+/-1.7 mm and 6.6+/-2.6mm maxilla; -0.4+/-1.4 mm and 3.2+/-1.5 mm mandible), and between ABC and CEJ (-3.4+/-1.3mm and 1.8+/-2.7 mm maxilla; -3.5+/-1.1mm and -0.1+/-1.4mm mandible) are significantly lower in test than in control animals. In line with these findings, periodontal attachment was 67% in the maxilla and 86% in the mandible of the test group and almost completely preserved in the control group. Clinical evaluation showed that the overall number of teeth with recessions was significantly higher in the test compared to the control group (4.9+/-2.4 and 2.3+/-3.6), but not the number of teeth missing and teeth with a probing depth>4mm. CONCLUSIONS: Together these findings suggest that in sheep, the cumulating effects of ageing, hypogonadism, malnutrition and glucocorticoid application can cause substantial catabolic changes of the periodontium. PMID- 19560121 TI - Dorsolateral prefrontal cortex activity predicts responsiveness to cognitive behavioral therapy in schizophrenia. AB - BACKGROUND: Given the variable response to cognitive-behavioral therapy (CBT) when added to antipsychotic medication in psychosis and the evidence for a role of pretherapy level of frontal lobe-based cognitive function in responsiveness to CBT in other disorders, this study examined whether pretherapy brain activity associated with working memory neural network predicts clinical responsiveness to CBT in schizophrenia. METHODS: Fifty-two outpatients stable on medication with at least one distressing symptom of schizophrenia and willing to receive CBT in addition to their usual treatment and 20 healthy participants underwent functional magnetic resonance imaging during a parametric n-back task. Subsequently, 26 patients received CBT for psychosis (CBT+treatment-as-usual [TAU], 19 completers) for 6-8 months, and 26 continued with TAU alone (17 completers). Symptoms in both patient groups were assessed (blindly) at entry and follow-up. RESULTS: The CBT+TAU and TAU-alone groups did not differ clinically or in performance at baseline. The CBT+TAU group showed significant improvement in relation to the TAU-alone group, which showed no change, at follow-up. Stronger dorsolateral prefrontal cortex (DLPFC) activity (within the normal range) and DLPFC-cerebellum connectivity during the highest memory load condition (2-back > 0-back) were associated with post-CBT clinical improvement. CONCLUSIONS: DLPFC activity and its connectivity with the cerebellum predict responsiveness to CBT for psychosis in schizophrenia. These effects may be mediated by PFC-cerebellum contributions to executive processing. PMID- 19560123 TI - Ventral striatal activation during reward anticipation correlates with impulsivity in alcoholics. AB - BACKGROUND: Alcohol dependence is often associated with impulsivity, which may be correlated with dysfunction of the brain reward system. We explored whether functional brain activation during anticipation of incentive stimuli is associated with impulsiveness in detoxified alcoholics and healthy control subjects. METHODS: Nineteen detoxified male alcoholics and 19 age-matched healthy men participated in a functional magnetic resonance imaging (fMRI) study using a monetary incentive delay (MID) task, in which visual cues predicted that a rapid response to a subsequent target stimulus would either result in monetary gain, avoidance of monetary loss, or no consequence. Impulsivity was assessed with the Barratt Impulsiveness Scale-Version 10 (BIS-10). RESULTS: Detoxified alcoholics showed reduced activation of the ventral striatum during anticipation of monetary gain relative to healthy control subjects. Low activation of the ventral striatum and anterior cingulate during gain anticipation was correlated with high impulsivity only in alcoholics, not in control subjects. CONCLUSIONS: This study suggests that reduced ventral striatal recruitment during anticipation of conventional rewards in alcoholics may be related to their increased impulsivity and indicate possibilities for enhanced treatment approaches in alcohol dependence. PMID- 19560124 TI - Ethylenesulfide as a useful agent for incorporation into the biopolymer chitosan in a solvent-free reaction for use in cation removal. AB - Chitosan (Ch) was chemically modified with ethylenesulfide (Es) under solvent free conditions to give (ChEs), displaying a high content of thiol groups due to opening of the three member cyclic reagent. Elemental analysis showed a decrease in nitrogen content. This result indicated the incorporation of two ethylenesulfide molecules for each unit of the polymeric structure of the precursor biopolymer. Infrared spectroscopy, thermogravimetry, and (13)C NMR in the solid state demonstrated the effectiveness of the reaction, with signals at 30 ppm for ChEs due to the change in the methylene group environment. Divalent metal uptake by chemically modified biopolymer gave the order Cu>Ni>Co>Zn, reflecting the corresponding acidity of these cations in bonding to the sulfur and the basic nitrogen atoms available on the pendant chains. The equilibrium data were fitted to Freundlich, Temkin, and Langmuir models. The maximum monolayer adsorption capacity for the cations was found to be 1.54+/-0.02, 1.25+/ 0.03, 1.13+/-0.01, and 0.83+/-0.03 mmol g(-1), respectively. The Langmuir model best explained the cation-sulfur bond interactions at the solid-liquid interface. The thermodynamics for these interactions gave exothermic enthalpic values of 43.02+/-0.03, -28.72+/-0.02, -26.27+/-0.04, and -17.32+/-0.02 kJ mol(-1), respectively. The spontaneity of the systems is given by negative Gibbs free energies of -31.2+/-0.1, -32.7+/-0.1, -31.7+/-0.1, and -32.2+/-0.1 kJ mol(-1), respectively, in spite of the unfavorable negative entropic values of -39+/-1, 13+/-1, -18+/-1, and -49+/-1 J K(-1)mol(-1) due to solvent ordering in the course of complexation. This newly synthesized biopolymer is presented as a chemically useful material for cation removal from aqueous solution. PMID- 19560122 TI - Childhood sexual abuse is associated with reduced gray matter volume in visual cortex of young women. AB - BACKGROUND: Childhood sexual abuse (CSA) has been associated with alterations in brain morphology using region of interest analyses that have focused on stress sensitive target regions. This study was designed to ascertain the effects on gray matter volume (GMV) of exposure to CSA in healthy young adult college students selected based on exposure history regardless of psychiatric outcome. Voxel-based morphometry (VBM) provided unbiased delineation of the most significantly affected brain regions. METHODS: High-resolution T1-weighted magnetic resonance imaging (MRI) datasets were obtained for 23 unmedicated female subjects with CSA and 14 healthy female control subjects of equivalent age and socioeconomic status with no history of trauma. Cortical surface-based analysis (FreeSurfer) was performed to verify VBM results. RESULTS: Gray matter volume was reduced by 12.6% and 18.1% in right and left primary visual (V1) and visual association cortices of abused subjects. This reduction was directly related to duration of CSA before age 12. Gray matter volume of left and right V1 correlated with measure of visual memory (r = .353, p = .032 and r = .448, p = .005). Cortical surface-based analysis indicated that GMV of abused subjects was reduced in the left fusiform (p = .004), left middle occipital (p = .04), and right lingual (p = .002) gyri. CONCLUSIONS: Early visual experience exerts a strong influence on the developing mammalian visual cortex. Present findings indicate that exposure to CSA may also affect the development of this region and are apparent even in a population of subjects who are sufficiently healthy to matriculate. PMID- 19560125 TI - Study on systematizing the synthesis of the a-series ganglioside glycans GT1a, GD1a, and GM1 using the newly developed N-Troc-protected GM3 and GalN intermediates. AB - A first systematic synthesis of the glycan parts of the a-series gangliosides (GT1a, GD1a, and GM1) utilizing the newly developed N-Troc-protected GM3 and galactosaminyl building blocks is described. The key processes, including the assembly of the GM2 sequence and its conversion into the 3-hydroxy acceptor, were facilitated mainly by the high degree of participation and chemoselective cleavability of the Troc group in the galactosaminyl unit. Furthermore, the novel GM2 acceptor served as a good coupling partner during glycosylation with galactosyl, sialyl galactosyl, and disialyl galactosyl donors, successfully producing the GM1, GD1a, and GT1a glycans. PMID- 19560126 TI - En route to deoxygenated N-acetyllactosamine analogues employing uridyl and galactosyl transferases. AB - All monodeoxygenated galactoses were treated with galactokinase, and for the 2-, 3-, and 4-deoxy compounds, transformation into the corresponding galactopyranosyl phosphates could be observed. In case of the 2-deoxy derivative, further reaction via UDP-2-deoxy-D-lyxo-hexose (UDP-2-deoxygalactose), which was also obtained chemically, the multiple enzymatic system could be employed to prepare 2'-deoxy-N acetyllactosamine. PMID- 19560127 TI - CpG oligodeoxynucleotides promote phospholipase D dependent phagolysosome maturation and intracellular mycobacterial killing in M. tuberculosis infected type II alveolar epithelial cells. AB - CpG oligodeoxynucleotides have been previously shown to enhance antimycobacterial response in human monocytes/macrophages. The present study reports evidences showing the capability of CpG oligodeoxynucleotides to induce (i) host phospholipase D (PLD) activation, (ii) PLD dependent reactive oxygen intermediate production, (iii) PLD dependent phagolysosome maturation and (iv) PLD dependent intracellular mycobacterial killing in type II alveolar epithelial cells. These are the first evidences showing that alveolar epithelial cells may represent efficient effecter cells during primary innate antimycobacterial immune response. PMID- 19560129 TI - [Cystocolic fistula secondary to peritoneal hydatidosis]. PMID- 19560128 TI - [Splenic abscess in an intravenous drug user: resolution with ultrasound-guided drainage]. PMID- 19560130 TI - [Vaginal tunic metastasis as a first sign of colorectal cancer]. PMID- 19560131 TI - On tickling brains to investigate minds. PMID- 19560132 TI - 'Normal' semantic-phonemic fluency discrepancy in Alzheimer's disease? A meta analytic study. AB - In a meta-analysis of 135 studies involving 6000 patients with Alzheimer's disease (AD) and 6057 healthy controls, we examined the relative degree of semantic and phonemic fluency impairment in AD patients. The effect size for semantic fluency (d=2.10: 95%CI 2.22-1.97) was significantly larger than for both phonemic fluency (d=1.46: 95%CI 1.56-1.36) and picture naming (d=1.54: 95%CI 1.66 1.40). In meta-regression analyses we found that studies with greater proportions of female patients and less severe dementia both led to better phonemic fluency; while perhaps surprisingly, increased patient education led to worse semantic fluency. Critically, in 50 studies measuring both semantic and phonemic fluency, the effect size for the semantic-phonemic discrepancy scores did not differ between AD patients and controls; and was unrelated to any of the moderator variables. The latter findings indicate that the semantic-phonemic fluency discrepancy measure often reported as an important distinguishing characteristic of AD patients may be an exaggerated normal tendency. PMID- 19560133 TI - Age-related differences in brain regions supporting successful encoding of emotional faces. AB - In an event-related functional Magnetic Resonance Imaging (fMRI) study, younger and older adults were presented with negative emotional (i.e., fearful) and neutral face pictures under incidental learning conditions. They were subsequently given a test of face recognition outside the scanner. Both age groups activated amygdala bilaterally as well as the right hippocampus during successful encoding of the fearful faces. Direct age comparisons revealed greater activation in right amygdala and bilateral hippocampus in the young, whereas older adults showed greater activation in the left insular and right prefrontal cortices. None of these brain areas was activated during successful encoding of neutral faces, suggesting specificity of these brain activation patterns. The results indicate an age-related shift in the neural underpinnings of negative emotional face processing from medial-temporal to neocortical regions. PMID- 19560134 TI - Cognitive reserve modulates task-induced activations and deactivations in healthy elders, amnestic mild cognitive impairment and mild Alzheimer's disease. AB - INTRODUCTION: Cognitive reserve (CR) reflects the capacity of the brain to endure neuropathology in order to minimize clinical manifestations. Previous studies showed that CR modulates the patterns of brain activity in both healthy and clinical populations. In the present study we sought to determine whether reorganizations of functional brain resources linked to CR could already be observed in amnestic mild cognitive impairment (a-MCI) and mild Alzheimer's disease (AD) patients when performing a task corresponding to an unaffected cognitive domain. We further investigated if activity in regions showing task induced deactivations, usually identified as pertaining to the default-mode network (DMN), was also influenced by CR. METHODS: Fifteen healthy elders, 15 a MCI and 15 AD patients underwent functional magnetic resonance imaging (fMRI) during a speech comprehension task. Differences in the regression of slopes between CR proxies and blood-oxygen-level dependent (BOLD) signals across clinical groups were investigated for activation and deactivation areas. Correlations between significant fMRI results and a language comprehension test were also computed. RESULTS: Among a-MCI and AD we observed positive correlations between CR measures and BOLD signals in task-induced activation areas directly processing speech, as well as greater deactivations in regions of the DMN. These relationships were inverted in healthy elders. We found no evidence that these results were mediated by gray matter volumes. Increased activity in left frontal areas and decreased activity in the anterior cingulate were related to better language comprehension in clinical evaluations. CONCLUSIONS: The present findings provide evidence that the neurofunctional reorganizations related to CR among a MCI and AD patients can be seen even when considering a preserved cognitive domain, being independent of gray matter atrophy. Areas showing both task-induced activations and deactivations are modulated by CR in an opposite manner when considering healthy elders versus patients. Brain reorganizations facilitated by CR may reflect behavioral compensatory mechanisms. PMID- 19560136 TI - Methods for defecting sperm apoptosis. PMID- 19560137 TI - Covered Wallstents for palliation of malignant biliary obstruction: primary stent placement versus reintervention. AB - BACKGROUND: Covered self-expandable metallic stents (SEMSs) are designed to prevent tumor ingrowth and can be removed if necessary. Only limited comparative data are available on the performance of covered SEMSs after primary placement versus reintervention. OBJECTIVE: To assess the effectiveness and safety of covered SEMS placement either as primary treatment or reintervention in patients requiring palliation of malignant biliary obstruction. DESIGN: Retrospective clinical cohort study. SETTING: Tertiary referral center. PATIENTS: This study involved 104 patients with unresectable malignant biliary strictures. INTERVENTION: Covered biliary SEMS placement. MAIN OUTCOME MEASUREMENTS: Stent patency, technical success, and patient survival. RESULTS: Covered SEMSs were placed as primary treatment in 48 patients (46%), and reintervention was performed in 56 patients (54%). At 3, 6, and 12 months thereafter, the Kaplan Meier estimated fractions of all patients with patent stents were 94%, 84%, and 58%, respectively. Covered SEMSs remained patent until the patient's death in 75 of 89 nonsurvivors (84%). Although patency rates 3, 6, and 12 months after primary placement (100%, 93%, and 82%, respectively) were higher than those after reintervention (90%, 78%, and 48%, respectively), the differences were not statistically significant (P = .057). Overall, the most frequent adverse events were cholangitis (7%) and stent migration (4%). LIMITATIONS: The distribution of stricture locations differed among the groups, and survival data suggested the presence of more extensive disease in the primary treatment group at baseline. CONCLUSION: The clinical utility and safety of primary covered SEMS placement were confirmed. This study provides the most extensive evidence to date that reintervention with a covered SEMS can provide a useful palliative option. PMID- 19560138 TI - EUS-guided radiofrequency ablation with a prototype electrode array system in an animal model (with video). AB - BACKGROUND: Although previously reported in an animal model, the development of EUS-guided radiofrequency ablation (EUS-RFA) has been impeded because of a lack of a retractable needle electrode array that could safely and effectively ablate large areas. OBJECTIVE: To evaluate the feasibility and safety of performing EUS RFA with a 19-gauge FNA needle fitted with an umbrella-shaped retractable needle electrode array. DESIGN AND SETTING: Endoscopic experimental study in a porcine survival model at a tertiary referral center animal laboratory. MAIN OUTCOME MEASURES: Evaluate the safety and efficacy of the retractable needle electrode array for performing EUS-RFA. INTERVENTIONS: A 19-gauge EUS-FNA needle was modified and fitted with a retractable echogenic umbrella-shaped monopolar electrode array at its tip. The FNA needle was connected to a 200-W generator that has an impedance-based feedback system. EUS-RFA of the liver was attempted on 5 Yorkshire pigs. Although 1 pig was euthanized immediately after RFA to assess for immediate complications and pathological examination, the 4 others were kept alive for 7 days. RESULTS: At EUS, the needle electrode was well visualized and could be deployed in the liver without technical difficulty. During ablation, a round hyperechoic focus gradually surrounded the electrode tip. Tissue ablation was attained within 7 minutes, and the electrode array could be easily withdrawn into the needle assembly. The vital signs of all pigs remained stable throughout the procedure and until they were euthanized. Histopathology in all pigs revealed a discrete, well-demarcated spherical focus of complete coagulation necrosis measuring 2.6 cm in diameter and without damage to the surrounding liver parenchyma or vasculature. CONCLUSIONS: In this experimental study, EUS-RFA of the liver was performed safely by using the retractable umbrella-shaped electrode array with effective coagulation necrosis of large areas. PMID- 19560139 TI - EUS-guided FNA aspiration of kidney masses: a multicenter U.S. experience. AB - BACKGROUND: Tissue sampling of renal lesions is traditionally performed with percutaneous US or CT guidance. To date, only 3 known cases of EUS-guided FNA (EUS-FNA) of a renal mass have been reported. OBJECTIVE: To describe a multicenter experience with the indications, yield, and complications from attempted EUS-FNA of a kidney mass. DESIGN: Retrospective case series. SETTING: Six tertiary referral hospitals in the United States. PATIENTS: Consecutive subjects undergoing attempted EUS-FNA of a kidney mass. Endosonographers at 15 other teaching hospitals were contacted regarding EUS findings and follow-up of any EUS-guided renal biopsies previously attempted or considered at that institution. INTERVENTIONS: EUS-FNA of a kidney mass. MAIN OUTCOME MEASUREMENTS: Biopsy indications, yield, diagnosis, and complications. RESULTS: Fifteen procedures in 15 patients (9 men; median age 67 years) were performed at 6 (37%) of 16 hospitals (Indiana University plus 15 other hospitals). Kidney masses (median diameter 32 mm; range 11-60 mm) were located in the upper (n = 12) and lower (n = 3) poles of the left (n = 10) and right (n = 5) kidneys, respectively. Initial mass detection was by previous imaging in 13 (87%) patients or by EUS in 2 (13%) patients. Results of EUS-FNA (median 3 passes; range 2-4 passes) in 13 (87%) procedures were diagnostic of (n = 7) or highly suspicious for (n = 1) renal cell carcinoma (RCC), atypical cells (n = 2), oncocytoma (n = 1), benign cyst (n = 1), and nondiagnostic (n = 1). No complications were encountered. Surgical resection confirmed RCC in 7 patients in whom preoperative EUS-FNA demonstrated RCC (n = 5) or oncocytoma (n = 1) or was not performed (n = 1). LIMITATIONS: Retrospective series, small number of patients. CONCLUSIONS: EUS-FNA of renal masses is rarely performed at the U.S. teaching hospitals surveyed. This technique appears safe and feasible and should be considered when results would affect patient management. PMID- 19560140 TI - EUS-guided fiducial placement before targeted radiation therapy for prostate cancer. AB - BACKGROUND: Image-guided radiation therapy allows the delivery of precisely aimed radiation beams to tumors while minimizing radiation to adjacent normal tissue. This is particularly important in the prostate, a moving target whose positioning depends on the dynamics of its neighboring bladder and rectum. Targeted radiation therapy can be achieved by using implantable radiographic markers, or fiducials, which serve as reference points to accurately delineate tumors. OBJECTIVE: To determine the feasibility and safety of placing fiducials in the prostate under linear array EUS guidance to facilitate targeted radiation therapy. DESIGN: Retrospective analysis of a prospective database. SETTING: University of Miami Hospital and Clinics, a tertiary cancer referral center. PATIENTS: Localized prostate cancer patients scheduled to undergo intensity-modulated radiation therapy. INTERVENTIONS: A total of 16 patients underwent EUS-guided fiducial placement to delineate the prostate before planned radiation therapy. RESULTS: Fiducial placement was successful in all patients (100%). A total of 71 gold markers were deployed in a 4-quadrant manner outlining the prostate. Seven of 16 patients had an additional fiducial placed to ensure adequate prostate delineation. Patients tolerated the procedure well with minimal discomfort. No complications developed from the procedure. LIMITATIONS: Single-center experience, small sample size. CONCLUSIONS: EUS-guided placement of fiducials to facilitate image-guided radiation therapy for prostate cancer is a feasible alternative to transperineal or transrectal US approaches, thereby adding to the expanding list of indications for linear EUS. This procedure can be safely performed by endosonographers familiar with perirectal anatomy and transrectal FNA technique. PMID- 19560141 TI - Placement of a self-expandable metal stent in a case of malignant stoma stenosis. PMID- 19560142 TI - A prospective evaluation of the feasibility of primary screening with unsedated colonoscopy. AB - BACKGROUND: Colonoscopy is the most effective screening tool for colorectal cancer. In Taiwan, colonoscopy is used much less than sigmoidoscopy for screening because sedation significantly increases the cost and is not readily available, and unsedated colonoscopy is considered to be poorly tolerated. However, unsedated colonoscopy has been shown to be well accepted and may improve the cost effectiveness and access to colonoscopic screening. OBJECTIVES: To compare the feasibility of unsedated colonoscopy and sigmoidoscopy for primary screening and to analyze factors associated with acceptance of the procedures and need for sedation. DESIGN: Single center, prospective. SETTING: National Taiwan University Medical Center. POPULATION AND INTERVENTIONS: A consecutive series of 261 subjects without history of colonoscopy or sigmoidoscopy who underwent unsedated colonoscopy (n = 176) or sigmoidoscopy (n = 85) for primary screening. MAIN OUTCOME MEASUREMENTS: Pain scores, acceptance, and need for sedation. RESULTS: No significant differences in pain, acceptance, and need for sedation were found between the colonoscopy and sigmoidoscopy groups. Only 9.6% in the colonoscopy group and 10.1% in the sigmoidoscopy group considered sedation necessary. Multivariate analyses revealed that the examinee's sex and the endoscopist, but not the type of endoscopic examination, were associated with the severity of pain and need for sedation. LIMITATIONS: Nonrandomized study design. CONCLUSIONS: Unsedated colonoscopy for primary screening is well accepted in nine tenths of examinees who accept this option and is similar to sigmoidoscopy in pain, acceptance, and need for sedation. Primary screening with unsedated colonoscopy is feasible, as with sigmoidoscopy. PMID- 19560143 TI - Linear enamel hypoplasia at Xcambo, Yucatan, during the Maya Classic period: An evaluation of coastal marshland impact on ancient human populations. AB - Non-specific stress markers such as linear enamel hypoplasia (LEH) have been associated in the literature with a large number of possible conditions disrupting the individual's homeostasis, though metabolic strain originating synergistically by disease and malnutrition has been held to be the main cause behind enamel disruption. The analysis of LEH in the Maya Classic period site of Xcambo, located along the northern coast of the Yucatan peninsula, reveals high exposure to stressful conditions during infancy regardless of age and sex. Yet, the inhabitants of the site were of a medium to high social and economic status, with access to balanced and protein-rich nutritional resources, which should have functioned as a cultural buffer to the impact of stress. In the light of this apparent contradiction, this paper discusses the impact of environmental conditions on the record of metabolic stress. Our conclusions pose a cautionary caveat for inferring nutrition and status in ancient pre-antibiotic populations solely from the occurrence of linear enamel hypoplasia. PMID- 19560144 TI - Adaptive NN output-feedback stabilization for a class of stochastic nonlinear strict-feedback systems. AB - In this paper, the adaptive neural network output-feedback stabilization problem is investigated for a class of stochastic nonlinear strict-feedback systems. The nonlinear terms, which only depend on the system output, are assumed to be completely unknown, and only an NN is employed to compensate for all unknown upper bounding functions, so that the designed controller is more simple than the existing results. It is shown that, based on the backstepping method and the technique of nonlinear observer design, the closed-loop system can be proved to be asymptotically stable in probability. The simulation results demonstrate the effectiveness of the proposed control scheme. PMID- 19560145 TI - Further validation of the Chinese version of the Level of Expressed Emotion Scale for research and clinical use. AB - BACKGROUND: Expressed emotion is a construct that has been used for the past three decades to describe family members' criticism, hostility and emotional involvement with a mentally ill relative within the context of family interactions and caregiving. In Western countries this construct is used as an outcome measure of interventions for families of people with schizophrenia or other psychotic disorders, but the use of this construct in Chinese populations is somewhat limited. OBJECTIVE: To test the reliability and validity of a refined Chinese version of the 52-item Level of Expressed Emotion Scale (LEE). METHODS: A convenience sample of 405 outpatients with psychotic disorders in Hong Kong and one of their family caregivers were recruited. Patients were asked to complete a set of questionnaires twice over a 6-month period, including the Chinese version of the LEE, the Specific Level of Functioning scale and the Brief Psychiatric Rating Scale; while their caregivers completed the Family Assessment Device twice and a demographic data sheet at recruitment. This study was aimed at establishing the internal consistency, reproducibility, responsiveness, and construct validity of the LEE. RESULTS: Results indicated that the refined 50-item Chinese version of the LEE and its subscales demonstrated a high internal consistency and satisfactory correlations with patient and family functioning scores. Principal component analysis revealed the presence of four factors, explaining 70.8% of total variance and indicating high factor loadings as well as item-factor inter correlations. The Chinese version not only indicated a satisfactory reproducibility in assessing change in patients' symptom severity and family functioning but also showed an adequate responsiveness to the changes in patients' symptoms over 6 months, especially for detecting symptom improvement. DISCUSSION: The findings of the psychometric evaluation of the Chinese version of the LEE established its potential as a research instrument in measuring the level of expressed emotion of family members as perceived by Chinese patients with psychotic disorders. Further testing of its psychometric properties is recommended, using larger samples from diverse socio-economic backgrounds and mental illnesses. PMID- 19560146 TI - Coronary heart disease and menopause management: the swinging pendulum of HRT. AB - The Women's Health Initiative comprised a randomized placebo-controlled clinical trial of therapeutic and dietary interventions influencing postmenopausal women's health. One arm evaluated hormone replacement therapy and its effects on major health outcomes. Initial publication of the preliminary results suggested overall harm from hormone replacement therapy, leading to a dramatic worldwide decrease in its use, and concerns from clinicians and regulatory authorities. Subsequent publications with more detailed analyses appear to have countermanded these initial concerns. Analyses of the studies have not been adherent to those specified in the original published protocol. Nominal confidence intervals should have been used only for the primary outcome, which was coronary heart disease. Initially reported as showing a significant increase in events with hormone replacement therapy, in a subsequent analysis of the full data the increase was no longer significant. Adjusted confidence intervals showed no significant increase for breast cancer, the primary adverse outcome. A major difference in the effects of hormones between younger and older women has emerged but this important finding has been minimized. For women under age 60 years or within 10 years of menopause, the final findings for all outcomes closely resemble those from observational cohorts. The raw data must be made available for independent assessment to obtain valid conclusions which may again change clinical practice. PMID- 19560147 TI - A response to the letter regarding the pleiotropic effects of fluvastatin on acute inflammatory response and the role of MMP-9 in plaque destabilization and intraplaque hemorrhage. PMID- 19560148 TI - Effects of body configuration on pelvic injury in backward fall simulation using 3D finite element models of pelvis-femur-soft tissue complex. AB - Injuries due to backward fall apart from sideways fall are a major health problem, particularly among the aged populations. The objectives of this study was to evaluate the responses to changing body configurations (angle between the trunk and impacting floor as 0 degrees, 15 degrees, 45 degrees and 80 degrees) during backward fall, based on a previously developed CT-scan-derived 3D non linear and non-homogeneous finite element (FE) model of pelvis-femur-soft tissue complex with simplified biomechanical representation of the whole body. Under constant impact energy, these FE models evaluated the pelvic injury situations on the basis of peak impact force (7.64-16.74 kN) and peak principal compressive strain (more than 1.5%), consistent with the clinically observed injuries (sacral insufficiency, coccydynia). Also the change in location of peak strain and increase in peak impact force for changing configurations from 0 degrees to 80 degrees indicated the effect of whole body inertia during backward fall. It was also concluded that the inclusion of sacro-iliac and acetabular cartilages in the above FE models will further reduce above findings marginally (9.2% for 15 degrees fall). These quantifications would also be helpful for a better design and development of safety structures such as safety floor for the nursing home or home for the aged persons. PMID- 19560149 TI - Analysis of pharmaceuticals in indirect potable reuse systems using solid-phase extraction and liquid chromatography-tandem mass spectrometry. AB - A solid-phase extraction (SPE) LC-MS/MS method for 18 commercial drugs in secondary wastewater and product water from water recycling plants using microfiltration (MF) and reverse osmosis (RO) has been developed, optimised and validated. The method incorporates a range of multi-class pharmaceuticals including lipid lowering agents, analgesics, antipyretics, non-steroidal anti inflammatory drugs, antidepressants, anticoagulants, tranquilizers, cytostatic agents, and antiepileptics. Method limits of quantitation (MLQs) in secondary wastewater ranged from 15 to 250 ng/L, while MLQs in post-RO water ranged from 1 to 25 ng/L. Results from analysis of secondary wastewater from Western Australia are presented, and represent the largest survey of non-antibiotic pharmaceuticals within Australia to date. Analysis of post-RO water from two MF/RO water recycling facilities also demonstrate that MF/RO treatment removes most pharmaceuticals to below the analytical limits of detection, and more importantly, up to seven orders of magnitude below health-based guideline values. PMID- 19560150 TI - Selectivity comparisons of monolithic silica capillary columns modified with poly(octadecyl methacrylate) and octadecyl moieties for halogenated compounds in reversed-phase liquid chromatography. AB - Stationary phase selectivities for halogenated compounds in reversed-phase HPLC were compared using C18 monolithic silica capillary columns modified with poly(octadecyl methacrylate) (ODM) and octadecyl moieties (ODS). The preferential retention of halogenated benzenes on ODM was observed in methanol/water and acetonitrile/water mobile phases. In selectivity comparison of selected analytes on ODM and ODS, greater selectivities for halogenated compounds were obtained with respect to alkylbenzenes on an ODM column, while similar selectivities were observed with a homologous series of alkylbenzenes on ODM and ODS columns. These data can be explained by greater dispersive interactions by more densely packed octadecyl groups on the ODM polymer coated column together with the contribution of carbonyl groups in ODM side chains. For the positional isomeric separation of dihalogenated benzenes (ortho-, meta-, para-), the ODM column also provided better separation of these isomers for the adjacently eluted isomers that cannot be completely separated on an ODS column in the same mobile phase. These results imply that the ODM column can be used as a better alternative to the ODS column for the separation of other halogenated compounds. PMID- 19560151 TI - Qualitative detection of diuretics and acidic metabolites of other doping agents in human urine by high-performance liquid chromatography-tandem mass spectrometry: comparison between liquid-liquid extraction and direct injection. AB - Direct injection of urine has gained interest in the field of analytical toxicology, including doping control analysis. However, implementation of a direct urinalysis method for the LC-MS/MS detection of 34 diuretics and 9 other doping agents yielded several analytical problems, which were not observed using a traditional liquid-liquid extraction. Therefore a comparative study was made between liquid-liquid extraction and direct injection. Comparison of validation results showed that the liquid-liquid extraction at pH 7 allows to analyze samples without major drawbacks regarding matrix effects. Hence, good sensitivity was observed and detection limits ranged between 1 and 250 ng/mL for all compounds. In the direct injection approach shifted retention times were observed for several acidic and basic compounds due to unwanted matrix effects. This shift was reduced by a 25-fold dilution of the urine samples. Besides the improved retention time stability the diluted samples also exhibited lower ion suppression than the undiluted ones. After 25-fold dilution, detection limits ranged between 10 and 250 ng/mL for all compounds. Since these detection limits are at or below the minimum required performance level, imposed by the World Anti-Doping Agency, the method could be applied to routine anti-doping analysis. Samples, previously declared positive, were reanalysed using both the liquid-liquid extraction and direct injection. With both techniques all 26 samples were found to be positive, showing the applicability of direct injection for the analysis of diuretics. PMID- 19560152 TI - Stability of binary and ternary model oil-field particle suspensions: a multivariate analysis approach. AB - The transmission profiles of one- to three-component particle suspension mixtures were analyzed by multivariate methods such as principal component analysis (PCA) and partial least-squares regression (PLS). The particles mimic the solids present in oil-field-produced water. Kaolin and silica represent solids of reservoir origin, whereas FeS is the product of bacterial metabolic activities, and Fe(3)O(4) corrosion product (e.g., from pipelines). All particles were coated with crude oil surface active components to imitate particles in real systems. The effects of different variables (concentration, temperature, and coating) on the suspension stability were studied with Turbiscan LAb(Expert). The transmission profiles over 75 min represent the overall water quality, while the transmission during the first 15.5 min gives information for suspension behavior during a representative time period for the hold time in the separator. The behavior of the mixed particle suspensions was compared to that of the single particle suspensions and models describing the systems were built. The findings are summarized as follows: silica seems to dominate the mixture properties in the binary suspensions toward enhanced separation. For 75 min, temperature and concentration are the most significant, while for 15.5 min, concentration is the only significant variable. Models for prediction of transmission spectra from run parameters as well as particle type from transmission profiles (inverse calibration) give a reasonable description of the relationships. In ternary particle mixtures, silica is not dominant and for 75 min, the significant variables for mixture (temperature and coating) are more similar to single kaolin and FeS/Fe(3)O(4). On the other hand, for 15.5 min, the coating is the most significant and this is similar to one for silica (at 15.5 min). The model for prediction of transmission spectra from run parameters gives good estimates of the transmission profiles. Although the model for prediction of particle type from transmission parameters is able to predict some particles, further improvement is required before all particles are consistently correctly classified. Cross-validation was done for both models and estimation errors are reported. PMID- 19560153 TI - Predicted models for phenanthrene sorption nonlinearity and capacity based on different HA/BC ratios in sediments. AB - Phenanthrene (Phen) sorption isotherms for synthetic sediment organic matter samples with nine different humic acid/black carbon (HA/BC) ratios were determined to provide predicted models for Phen sorption nonlinearity and affinity by sediments. The sorption data were fitted well by the Freundlich model. The nonlinear sorption of Phen was dominated by BC particles. According to BC/TOC content, Phen sorption nonlinearity can be predicted using the exponential model: n=0.410+0.454 *exp(-0.028 BC/TOC). As sorption is a combination of absorption into soft carbon and adsorption onto hard carbon, based on published K(HA), K(FBC), and n(bc) values and measured f(BC) and f(oc) values, Phen sorption affinity-related parameter K(oc) values can be predicted from the simplified dual sorption model: f(oc)K(oc) = (f(oc) - f(BC))K(HA) + f(BC)K(FBC)C(e)(n(bc) - 1). For sediments from the Song-Liao watershed (China), the exponential model and dual sorption model gave good predictions for n and K(oc) values, which support the availability of the two predicted models. PMID- 19560154 TI - Microstructures and rheological dynamics of viscoelastic solutions in a catanionic surfactant system. AB - Viscoelastic solutions formed in a catanionic surfactant system of dodecyltriethylammonium bromide (DTEAB)/sodium dodecylsulfate (SDS) at the molar ratio of 27/73 were systematically studied using a combination of rheology and dynamic light scattering (DLS). Wormlike micelles began to form above the total surfactant concentration (C(total)) of 120 mM by the growth of small cylindrical micelles. Subsequently the system was found to exhibit linear viscoelasticity with characteristic of a Maxwell fluid in the intermediate concentration range of 170-210 mM, which arose from a 3D entangled network of wormlike micelles. At higher surfactant concentrations, a transition from linear micelles to branched structures probably took place. Finally and significantly, the effect of the surfactant headgroup on the rheological property of catanionic surfactant mixtures was discussed. PMID- 19560155 TI - Cu-doped resorcinol-formaldehyde (RF) polymer and carbon aerogels. AB - Introduction of transition metal salt(s) onto the surface of porous carbons may increase the selectivity and/or efficiency of these adsorbents in catalysis or separation. Carbon aerogels with low pressure drop are particularly suited for these applications. Moreover the sol-gel process used in the synthesis of the resorcinol-formaldehyde polymer gel (RF) precursors offers an extra opportunity for introducing metal ions. Salts of different metals modify both the macroscopic texture and the porosity, depending on the synthesis protocol. In this paper we show, by means of low temperature nitrogen adsorption measurements and SEM, as well as small- and wide-angle X-ray scattering (SAXS and WAXS), how the addition of copper acetate at three different stages influences not only the specific surface area but also the resulting overall structure over a wide range of length scales. Posttreatment in either the polymer or the carbon aerogel stage provides a means of adjusting the copper content. While the Cu-containing carbon aerogels differ mainly in their micropore volume but not in the width of the distribution, their pore size window in the mesopore range can be tuned between 50 and 400 A by the protocol of Cu addition. The synthesis protocol also determines the chemical form of the copper. PMID- 19560156 TI - Comparison of gas-solid chromatography and MM2 force field molecular binding energies for greenhouse gases on a carbonaceous surface. AB - Gas-solid chromatography was used to determine B(2s) (gas-solid virial coefficient) values for eight molecular adsorbates interacting with a carbon powder (Carbopack B, Supelco). B(2s) values were determined by multiple size variant injections within the temperature range of 313-553 K. The molecular adsorbates included: carbon dioxide (CO(2)); tetrafluoromethane (CF(4)); hexafluoroethane (C(2)F(6)); 1,1-difluoroethane (C(2)H(4)F(2)); 1-chloro-1,1 difluoroethane (C(2)H(3)ClF(2)); dichlorodifluoromethane (CCl(2)F(2)); trichlorofluoromethane (CCl(3)F); and 1,1,1-trichloroethane (C(2)H(3)Cl(3)). Two of these molecules are of special interest because they are "super greenhouse gases". The global warming potential, GWP, for CF(4) is 6500 and for C(2)F(6) is 9200 relative to the reference value of 1 for CO(2). The GWP index considers both radiative blocking and molecular lifetime. For these and other industrial greenhouse gases, adsorptive trapping on a carbonaceous solid, which depends on molecule-surface binding energy, could avoid atmospheric release. The temperature variations of the gas-solid virial coefficients in conjunction with van't Hoff plots were used to find the experimental adsorption energy or binding energy values (E(*)) for each adsorbate. A molecular mechanics based, rough-surface model was used to calculate the molecule-surface binding energy (Ecal(*)) using augmented MM2 parameters. The surface model consisted of parallel graphene layers with two separated nanostructures each containing 17 benzene rings arranged in linear strips. The separation of the parallel nanostructures had been optimized in a prior study to appropriately represent molecule-surface interactions for Carbopack B. Linear regressions of E(*) versus Ecal(*) for the current data set of eight molecules and the same surface model gave E(*)=0.926 Ecal(*) and r(2)=0.956. A combined set of the current and prior Carbopack B adsorbates studied (linear alkanes, branched alkanes, cyclic alkanes, ethers, and halogenated hydrocarbons) gave a data set with 33 molecules and a regression of E(*)=0.991 Ecal(*) and r(2)=0.968. These results indicated a good correlation between the experimental and the MM2 computed molecule-surface binding energies. PMID- 19560157 TI - The role of abdominal radiography in the diagnosis of intussusception when interpreted by pediatric emergency physicians. AB - OBJECTIVE: To evaluate the sensitivity and specificity of abdominal x-rays in the diagnosis of intussusception when interpreted by pediatric emergency physicians. STUDY DESIGN: This was a prospective experimental study. Participants were board certified/eligible pediatric emergency physicians. They evaluated a module containing radiographs of 50 cases of intussusception and 50 controls, matched for age and sex. For each x-ray, the physicians stated whether the x-ray increased, decreased or did not affect suspicion of intussusception. The primary outcome was the percentage of cases for which physicians stated that the x-ray increased their level of suspicion (sensitivity). Secondary outcomes included the proportion of false-negative results and specificity. RESULTS: Fourteen of 15 eligible physicians participated in the study. Overall, abdominal radiography increased the index of suspicion of intussusception in 48% of cases (sensitivity) and 21% of controls; however, in 11% of cases, the abdominal x-rays were incorrectly interpreted as being reassuring. The specificity was 21%. The radiographs were deemed equivocal for 41% of cases and 58% of controls. CONCLUSIONS: Abdominal x-rays have a low sensitivity and specificity for diagnosing intussusception when interpreted by pediatric emergency physicians. PMID- 19560159 TI - The role of laparoscopy in the management of malrotation. AB - BACKGROUND: Malrotation is currently treated via the Ladd's procedure. Many surgeons feel this operation should be performed using the open approach to facilitate adhesion development, thus decreasing the risk for volvulus. However, little comparative data exist on the relative merits of laparoscopy for this operation. Therefore, we have analyzed our experience with the open and laparoscopic Ladd's procedure. METHODS: A retrospective analysis of our most recent 13-y experience with the Ladd's procedure was performed. Demographics, approach, diagnosis, hospital course, and outcomes were measured. Data are expressed as mean +/- standard deviation. Comparative analysis was performed using a t-test. RESULTS: A total of 284 Ladd's procedures were performed during this time, of which 43 were approached laparoscopically. Conversion rate was 33%, usually due to concern for volvulus/orientation. Volvulus after Ladd's procedure occurred in six patients, all of whom underwent an open approach (2.4%). Recovery data excluding patients who underwent bowel resection are displayed in Table 1. CONCLUSIONS: A laparoscopic Ladd's procedure should be the initial approach in patients with malrotation in the absence of volvulus. We encourage a low threshold for conversion to an open approach if there is any concern about volvulus/orientation. This may decrease morbidity for patients who are found at operation to have a low risk of recurrent volvulus. PMID- 19560158 TI - Mercury levels in premature and low birth weight newborn infants after receipt of thimerosal-containing vaccines. AB - OBJECTIVE: We conducted a population-based pharmacokinetic study to assess blood levels and elimination of mercury after vaccination of premature infants born at > or =32 and <37 weeks of gestation and with birth weight > or =2000 but <3000 g. STUDY DESIGN: Blood, stool, and urine samples were obtained before vaccination and 12 hours to 30 days after vaccination from 72 premature newborn infants. Total mercury levels were measured by atomic absorption. RESULTS: The mean +/- standard deviation (SD) birth weight was 2.4 +/- 0.3 kg for the study population. Maximal mean +/- SD blood mercury level was 3.6 +/- 2.1 ng/mL, occurring at 1 day after vaccination; maximal mean +/- SD stool mercury level was 35.4 +/- 38.0 ng/g, occurring on day 5 after vaccination; and urine mercury levels were mostly nondetectable. The blood mercury half-life was calculated to be 6.3 (95% CI, 3.85 to 8.77) days, and mercury levels returned to prevaccination levels by day 30. CONCLUSIONS: The blood half-life of intramuscular ethyl mercury from thimerosal in vaccines given to premature infants is substantially shorter than that of oral methyl mercury in adults. Because of the differing pharmacokinetics, exposure guidelines based on oral methyl mercury in adults may not be accurate for children who receive thimerosal-containing vaccines. PMID- 19560160 TI - Loss of airway pressure during HFOV results in an extended loss of oxygenation: a retrospective animal study. AB - BACKGROUND: Patients with acute respiratory distress syndrome (ARDS) are often ventilated with high airway pressure. Brief loss of airway pressure may lead to an extended loss of oxygenation. While using high frequency oscillatory ventilation (HFOV) in a porcine acute lung injury model, two animals became disconnected from the ventilator with subsequent loss of airway pressure. We compared the two disconnected animals to the two animals that remained connected to determine causes for the extended reduction in oxygenation. METHODS: ARDS was induced using 5% Tween. Thirty min of nonprotective ventilation (NPV) followed before placing the pigs on HFOV. Measurements were made at baseline, after lung injury, and every 30min during the 6-h study. Disconnections were treated by hand ventilation and a recruitment maneuver before being placed back on HFOV. The lungs were histologically analyzed and wet/dry weights were measured to determine lung edema. RESULTS: Hemodynamics and lung function were similar in all pigs at baseline, after injury, and following NPV. The animals that remained connected to the oscillator showed a continued improvement in PaO(2)/FiO(2) (P/F) ratio throughout the study. The animals that experienced the disconnection had a significant loss of lung function that never recovered. The disconnect animals had more diffuse alveolar disease on histologic analysis. CONCLUSIONS: A significant fall in lung function results following disconnection from HFOV, which remains depressed for a substantial period of time despite efforts to reopen the lung. Dispersion of edema fluid is a possible mechanism for the protracted loss of lung function. PMID- 19560161 TI - Sepsis and major abdominal surgery lead to flaking of the endothelial glycocalix. AB - BACKGROUND: Recent evidence suggests that the endothelial glycocalix plays an important role in lethal outcomes following sepsis. We therefore tested if the endothelial glycocalix is shed in patients with sepsis compared with patients after major abdominal surgery and healthy volunteers. MATERIAL AND METHODS: A total of 150 individuals were tested for levels of inflammatory markers (intercellular adhesion molecule-1 [ICAM-1], vascular cell adhesion molecule-1 [VCAM-1], interleukin-6 [IL-6]) and glycocalix markers (syndecan-1, heparan sulfate). Three groups consisted of patients with severe sepsis or septic shock, patients after major abdominal surgery without systemic inflammatory response syndrome, and healthy volunteers. Blood was drawn, at the time of diagnosis or surgery, and 6, 24, and 48h later. We correlated these markers to each other and to clinically used inflammation markers. RESULTS: Levels of inflammatory markers were markedly higher in patients with sepsis compared with patients after major abdominal surgery and healthy volunteers. After major abdominal surgery, glycocalix markers in human plasma were at levels comparable to patients with sepsis. In patients with sepsis, levels of IL-6 correlated with syndecan-1, ICAM 1, VCAM-1, and lactate, while ICAM-1 furthermore correlated with CRP and lactate levels. CONCLUSION: High levels of glycocalix markers indicated that significant flaking of the endothelial glycocalix occurred in patients with sepsis, and to a lesser extent in patients after major abdominal surgery. This novel finding could explain the nonspecific capillary leaking syndrome of patients with sepsis and after major abdominal surgery, and may identify new targets for treating those patient populations. PMID- 19560162 TI - Use of abdominal wall allotransplantation as an alternative for the management of end stage abdominal wall failure in a porcine model. AB - BACKGROUND: We describe a novel approach for treating end stage abdominal wall failure using isolated abdominal wall allotransplantation in a porcine model. METHODS: Full thickness abdominal wall transplants were performed in 13 pairs of genetically mismatched pigs. All recipients received daily immunosuppresion after transplantation. Rejection was assessed by visual inspection and skin biopsies. At the end of the 28 d study period, thickness, stiffness, and tensile strength of the transplanted rectus muscle was measured and compared with native rectus muscle. RESULTS: Eight grafts were viable and showed no signs of herniation. Four grafts failed within the first week secondary to vascular thrombosis. One animal had viable graft but was euthanized due to an incarcerated inguinal hernia. Rejection was minimal in six of the eight recipients. At necropsy, the gross thickness of the transplanted muscle flap was reduced compared with the native muscle (4.3 mm versus 7.7 mm, P < 0.001). Histologically, the diameter of the muscle fiber decreased from 0.15 mm to 0.09 mm (P < 0.0001). While the stiffness measurements between the transplanted and native muscles were comparable, the transplanted muscles had significantly lower tensile strength than the native muscles. CONCLUSION: This study demonstrates the feasibility of isolated abdominal wall allotransplantation to provide a potential solution for end stage abdominal wall failure. Based on the model set forth, future work will evaluate the biomechanical properties of the composite allograft to provide a suitable dynamic abdominal wall replacement. PMID- 19560163 TI - Pediatric intestinal foregut and small bowel solid tumors: a review of 105 cases. AB - BACKGROUND: The outcomes of pediatric intestinal foregut and small bowel solid tumors have never been studied on a population scale. MATERIALS AND METHODS: The Surveillance, Epidemiology, and End Results database (1973-2005) was queried for all patients under 20 y of age. RESULTS: A total of 105 cases of pediatric intestinal foregut and small bowel solid tumors were identified. Tumors occurred in the esophagus (8.6%), stomach (61%), and small bowel (30.5%). The most common histologies include sarcoma (43.8%), which consisted mostly of gastrointestinal stromal tumors (GIST), carcinoma (41.0%), which consisted mostly of adenocarcinomas, and neuroendocrine tumors (NET) (10.5%). Most tumors were poorly differentiated and presented with advanced disease. The overall median survival time was 207 mo. Gastric solid tumors had significantly worse 5- and 10-y survival compared with their small bowel counterparts, though this difference disappeared in those who received surgical resection. Patients with carcinoma had significantly worse survival compared with those with sarcoma or NET, regardless of site and surgical intervention. Univariate analysis identified race, differentiation, stage, and surgery as significant predictors of survival. Multivariate analysis revealed that African American race, advanced stage of disease, carcinoma histology, and failure to undergo surgical extirpation were all independent predictors of worse outcome. In patients with carcinoma, failure to undergo radiotherapy was also a predictor of worse outcome. CONCLUSION: Surgery is associated with a significantly improved survival for pediatric patients with solid tumors of the intestinal foregut and small bowel. Radiotherapy appears to be an important adjuvant therapy for patients with carcinoma. PMID- 19560164 TI - Clinical impact of segmentectomy compared with lobectomy under complete video assisted thoracic surgery in the treatment of stage I non-small cell lung cancer. AB - BACKGROUND: Segmentectomy for small or early stage non-small cell lung cancer (NSCLC) has been controversial. Further, video-assisted thoracic surgery (VATS) for lung cancer was accepted during the past decade. We here compared the outcome between VATS segmentectomy and VATS lobectomy for stage I NSCLC. METHODS: In the retrospective study, 109 consecutive patients in stage I underwent surgery at Oita University Hospital (Oita, Japan) between September 2003 and October 2008. VATS segmentectomy was performed in 38 patients and VATS lobectomy with systemic lymphadenectomy was performed in 71 patients. After clinicopathologic factors were compared in both groups, local recurrence rates and survivals were compared. RESULTS: Five of 38 VATS segmentectomy and eight of 71 VATS lobectomy patients relapsed during the follow-up period (median 27.5 mo). In the relapsed patients after VATS segmentectomy, three (7.9%) were local recurrences and two (5.3%) were distant metastases. On the other hand, four (5.6%) were local recurrence and four (5.6%) were distant metastases in the VATS lobectomy group. There was no significant difference between the two groups. Furthermore, there was no difference in recurrence-free and overall survival between segmentectomy and lobectomy. CONCLUSIONS: Although the sample size is small, VATS segmentectomy is one of the appropriate procedures for stage I NSCLC. PMID- 19560165 TI - Differential expression of procollagen genes between mid- and late-gestational fetal fibroblasts. AB - BACKGROUND: Previously, we have shown that cutaneous wounds in mid-gestational (E15) mice heal in a scarless manner with decreased procollagen 1 and increased procollagen 3 production compared with wounds in late-gestational (E18) mice, which heal with scars. The aim of the current work was to determine whether E15 and E18 fibroblasts respond to stimulation in culture with differential procollagen expression, suggesting they may preserve their phenotype in vitro. Further, we wanted to determine if fetal fibroblast gene expression patterns persisted in tissue culture. We measured expression of procollagen types 1alpha1 and 3 in response to TGF-beta1 stimulation. We theorized that E15 fibroblasts would respond with a pattern of procollagen that would contribute to a more easily remodeled collagen. METHODS: Mid- and late-gestational fetal fibroblasts were obtained from dorsal skin harvested from fetuses of time-dated CD-1 mice. Cells were grown to confluence in culture plates overnight. Cell monolayers were treated with 0.01% bovine serum albumin (BSA) plus 10 ng/well of TGF-beta1. Cells were harvested at 6 and 24 h following treatment. Additional groups were treated with BSA alone (vehicle controls) and collected at 6 and 24 h. Another group without treatment was harvested after reaching confluence (0 time point). In a separate experiment to determine if gene expression patterns persisted, cells were treated with 0.01% BSA plus 10 ng/well of TGF- beta1 for 24 h, then harvested. A second group of cells were treated again at 24 h and harvested at 48 h. Additional cells were treated with BSA alone for 24 and 48 h, and another group without treatment was harvested after reaching confluence (0 time point). Cells were processed to obtain mRNA, cDNA was made, and then samples analyzed by QPCR. Results were analyzed by ANOVA and Holm-Sidak method. RESULTS: Procollagen 1alpha1 gene expression was decreased in E15 cells at 6 and 24 h following TGF beta1 treatment, P<0.05. In contrast, procollagen 1alpha1 was increased in E18 cells, P<0.05. Procollagen 3 gene expression was decreased in E18 cells at 6 and 24 h following treatment with TGF-beta1, P<0.05, whereas levels in E15 cells were unchanged at 6 h, and only trended lower at 24 h. We evaluated whether this differential expression of procollagen 3 persisted at 24 and 48 h. At 24 and 48 h, E15 control groups had increased procollagen 3 expression compared with E18 groups, P<0.05. E15 and E18 cells in TGF-beta1-treated groups had decreased procollagen 3 at 48h compared with their respective BSA control groups, P<0.05. However, the degree of difference appeared to be greater in the E15 group than the E18 group. CONCLUSIONS: Our results from this in vitro work demonstrate a differential pattern of gene expression for procollagen 1alpha1 and 3 in E15 and E18 fibroblasts in response to TGF-beta1. E15 cells showed decreased expression of procollagen 1alpha1, while E18 cells showed increased procollagen 1alpha1 and decreased procollagen 3 expression. These patterns of expression in E15 cells are suggestive of increased type 3 to 1 collagen ratio seen in scarless fetal wounds. Interestingly, treatment of either E15 or E18 cells with TGF-beta1 significantly decreased procollagen 3 expression by 48 h, yet this was more profound in E15 groups. This suggests that after 24 h, E15 cells may transition towards an E18 phenotype and corresponding signaling. PMID- 19560166 TI - Evaluation of MRI for the diagnosis of appendicitis during pregnancy when ultrasound is inconclusive. AB - BACKGROUND: To retrospectively evaluate the diagnostic performance and clinical utility of magnetic resonance imaging (MRI) in pregnant patients suspected of having acute appendicitis, when an ultrasound study generated an inconclusive result. METHODS: The medical records of 19 consecutive women who underwent abdominal and pelvic MRI at a tertiary care referral center (St. Paul's Hospital, Vancouver, Canada), as part of the work up of clinically suspected acute appendicitis, were retrospectively reviewed. MRI was carried out when ultrasound findings were inconclusive. MRI findings were reviewed and compared with surgical findings and clinical follow-up data including pregnancy outcome. RESULTS: One of the 19 patients (5.3%) in the study cohort had an appendicitis diagnosed by MRI that was confirmed at operation and by specimen histology. The remaining study patients were diagnosed as not having appendicitis by MRI. These patients were followed until delivery, which was uneventful for all but one patient who was found to have appendicitis during Cesarean section. Overall, the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of MRI for the diagnosis of appendicitis during pregnancy was 50.0%, 100%, 100%, 94.4%, and 94.7%, respectively. In three patients (16.7%) with no MRI evidence of appendicitis, MRI identified an alternative etiology for their abdominal pain (two patients diagnosed with ovarian cysts, one patient diagnosed with a uterine fibroid). CONCLUSIONS: MRI represents a useful diagnostic test for acute appendicitis in pregnant women, and decreases the need for an emergency operation. Its high negative predictive value makes MRI useful for ruling out appendicitis in pregnant patients who have an inconclusive ultrasound. However, the low sensitivity observed in this study suggests that MRI, like other imaging modalities, is not perfect, and may miss an acute appendicitis diagnosis. Thus, future prospective clinical study of MRI as a diagnostic test for the evaluation of women who present with acute abdominal pain and possible appendicitis during pregnancy is warranted. PMID- 19560167 TI - Effect of oxygenated perfluorocarbon on isolated islets during transportation. AB - BACKGROUND: Previous studies demonstrated the efficacy of the two-layer method (TLM) using oxygenated perfluorochemicals (PFC) for pancreas preservation. The current study investigated the effect of oxygenated PFC on isolated islets during transportation. MATERIALS AND METHODS: Purified rat islets were stored in an airtight conical tube for 24h in RPMI culture medium at 22 degrees C or University of Wisconsin solution (UW) at 4 degrees C, either with or without oxygenated PFC. After storage, the islets were assessed for in vitro viability by static incubation (SI), FDA/PI staining, and energy status (ATP, energy charge, and ADP/ATP ratio) and for in vivo viability by a transplantation study. RESULTS: UW at 4 degrees C and RPMI medium at 22 degrees C maintained islet quality almost equally in both in vitro and in vivo assessments. The ATP levels and energy status in the groups with PFC were significantly lower than those without PFC. The groups with PFC showed a significantly higher ADP/ATP ratio than those without PFC. In the transplantation study, blood glucose levels and AUC in the UW+PFC group were significantly higher than those in UW group. CONCLUSIONS: UW at 4 degrees C and RPMI medium at 22 degrees C maintained islet quality equally under the conditions for islet transportation. The addition of oxygenated PFC, while advantageous for pancreas preservation, is not useful for islet transportation. PMID- 19560169 TI - Recent aquatic ecosystem response to environmental events revealed from 210Pb sediment profiles. AB - The (210)Pb dating method was first introduced by Goldberg (1963), and since then has been applied to study sediment from lakes, estuaries and coastal marine environments. Hundreds of studies around the world have used (210)Pb as a geochronological tool in aquatic ecosystems. However little attention has been paid to the potential of this naturally occurring isotope as an environmental tracer of ecological events. Here we report three instances in which (210)Pb profiles measured on undisturbed sediment cores from lakes, rivers and fjords show us the potential of (210)Pb profile as a tracer of natural and anthropogenic processes. The methodology used here is a suite of techniques combining biogeochemistry (micro-electrodes), paleomagnetism (susceptibility), sediment characteristics (LOI) and visualization (SPI and X-ray) applied to the interpretation of (210)Pb profiles. We measured (210)Pb profiles on sediments from a river, Cruces River (Chile), which recorded a clear shift in the water chemistry caused by a pulp mill effluent to the river. Here metal mobilization and remobilization of the tracer may be the cause of the observed profile. We also measured (210)Pb profiles in sediment from two fjords of Southern Chile (Pillan and Renihue), the sudden deposition change of fresh (210)Pb with depth observed could very well be the result of bioturbation but it occurred in a seafloor area deprived of bioturbators. In this case, (210)Pb recorded the onset of aquaculture activities (fish farming) that took place two decades ago. Finally, (210)Pb profiles measured in two lakes in the "pampa Argentina": Epecuen and Venado showed a particular shape with depth. These profiles apparently registered a sudden depositional event with recent (210)Pb material, probably related to strong shifts in precipitation and drought cycles in that part of the world. These three examples show that (210)Pb profiles provide valuable information not only on geochronology, but also related to natural and anthropogenic short term processes, as shown here, but these are not always reported and well understood. PMID- 19560168 TI - Multiple sclerosis associated fatigue during natalizumab treatment. AB - OBJECTIVE: To assess multiple sclerosis (MS) associated fatigue after the first 6 months of natalizumab treatment. METHODS: Prospective, open-label, uncontrolled study. Fatigue was measured before treatment initiation and at month 6 with the Modified Fatigue Impact Scale (MFIS) and the Fatigue Severity Scale (FSS) in 42 patients. Subjective impression of well-being was assessed with a visual analogue scale (1-10). RESULTS: 42 patients were followed (mean age 35.1 years, 60% female, mean EDSS 3.7). The mean total MFIS were 45.8 (17.5) at baseline and decreased to 40.1 (18.0) (p<0.01) at month 6. Mean scores on the VAS for subjective impression of well-being increased from 5.5 (1.9) to 6.1 (2.1) at month 6 (p<0.01). Pre-treatment annual relapse rate decreased from 2.2 to 0.2, gadolinium enhancing lesions were reduced by 96% with natalizumab. There was no correlation of gadolineum enhancing lesions and fatigue scores. CONCLUSION: Fatigue and well-being improved after treatment initiation with natalizumab. A randomized controlled trial is necessary to come to definite conclusions as to a potential effect of natalizumab on fatigue in MS. PMID- 19560170 TI - [Physical exercise and consumption of drugs for osteoarticular diseases in people older than sixty-five years]. PMID- 19560172 TI - [Development of the CAT-Health, the first adaptive computerized test (CAT) for the evaluation health-related quality of life in Spain]. AB - OBJECTIVES: To develop a Computerized Adaptive Test to obtain a generic measure of the patients' perceived health state. METHODS: An experts' panel created a pool of five-alternative Likert format items, chosen among Health Related Quality of Life questionnaires validated in our country. That pool was applied to a pilot sample and later to another general sample of patients belonging to 7 Primary Health Centers in Asturias. Two response options were offered to the general sample: 1) on paper 2) on a touch panel. The Item Response Theory psychometric properties (discrimination, reliability and validity) were evaluated by means of the factorial analysis and other methods. The information function was analyzed and a study of simulation was developed. RESULTS: A pool of 140 items was created. The requirements for the application of the IRT were verified by means of the factorial analysis. A selection of items was realized following the statistical criterion and the application of the IRT. The elimination of 35 articles resulted in a test composed by 105 items. Once the questionnaire was applied to the general sample, the parameters of the items were estimated again. The parameters invariance was verified once more and other 9 items were eliminated. The result was a definitive version of 96 items. CONCLUSIONS: The Computerized Adaptive Test developed proves to be a fast and reliable tool to evaluate the Health Related Quality of Life generically. This test may imply a very important advance for the measurement of the results reported by the patients in the clinical practice. PMID- 19560171 TI - [HIV-2 and HTLV-1/2 infections in Spain]. PMID- 19560173 TI - [Specific-type 2 diabetes mellitus health-related quality of life measurements available for Spanish population]. PMID- 19560174 TI - Diversity, regulation, and evolution of the gibberellin biosynthetic pathway in fungi compared to plants and bacteria. AB - Bioactive gibberellins (GAs) are diterpene plant hormones that are biosynthesized through complex pathways and control diverse aspects of growth and development. GAs were first isolated as metabolites of a fungal rice pathogen, Gibberella fujikuroi, since renamed Fusarium fujikuroi. Although higher plants and the fungus produce structurally identical GAs, significant differences in their GA pathways, enzymes involved and gene regulation became apparent with the identification of GA biosynthetic genes in Arabidopsis thaliana and F. fujikuroi. Recent identifications of GA biosynthetic gene clusters in two other fungi, Phaeosphaeria spp. and Sphaceloma manihoticola, and the high conservation of GA cluster organization in these distantly related fungal species indicate that fungi evolved GA and other diterpene biosynthetic pathways independently from plants. Furthermore, the occurrence of GAs and recent identification of the first GA biosynthetic genes in the bacterium Bradyrhizobium japonicum make it possible to study evolution of GA pathways in general. In this review, we summarize our current understanding of the GA biosynthesis pathway, specifically the genes and enzymes involved as well as gene regulation and localization in the genomes of different fungi and compare it with that in higher and lower plants and bacteria. PMID- 19560175 TI - Evolution of rosmarinic acid biosynthesis. AB - Rosmarinic acid and chlorogenic acid are caffeic acid esters widely found in the plant kingdom and presumably accumulated as defense compounds. In a survey, more than 240 plant species have been screened for the presence of rosmarinic and chlorogenic acids. Several rosmarinic acid-containing species have been detected. The rosmarinic acid accumulation in species of the Marantaceae has not been known before. Rosmarinic acid is found in hornworts, in the fern family Blechnaceae and in species of several orders of mono- and dicotyledonous angiosperms. The biosyntheses of caffeoylshikimate, chlorogenic acid and rosmarinic acid use 4 coumaroyl-CoA from the general phenylpropanoid pathway as hydroxycinnamoyl donor. The hydroxycinnamoyl acceptor substrate comes from the shikimate pathway: shikimic acid, quinic acid and hydroxyphenyllactic acid derived from l-tyrosine. Similar steps are involved in the biosyntheses of rosmarinic, chlorogenic and caffeoylshikimic acids: the transfer of the 4-coumaroyl moiety to an acceptor molecule by a hydroxycinnamoyltransferase from the BAHD acyltransferase family and the meta-hydroxylation of the 4-coumaroyl moiety in the ester by a cytochrome P450 monooxygenase from the CYP98A family. The hydroxycinnamoyltransferases as well as the meta-hydroxylases show high sequence similarities and thus seem to be closely related. The hydroxycinnamoyltransferase and CYP98A14 from Coleus blumei (Lamiaceae) are nevertheless specific for substrates involved in RA biosynthesis showing an evolutionary diversification in phenolic ester metabolism. Our current view is that only a few enzymes had to be "invented" for rosmarinic acid biosynthesis probably on the basis of genes needed for the formation of chlorogenic and caffeoylshikimic acid while further biosynthetic steps might have been recruited from phenylpropanoid metabolism, tocopherol/plastoquinone biosynthesis and photorespiration. PMID- 19560178 TI - [Radiologia and opinion]. PMID- 19560176 TI - Assessment of doctors' knowledge regarding tuberculosis management in Lucknow, India: a public-private sector comparison. AB - OBJECTIVE: India tops the list of 22 high-burden tuberculosis (TB) countries. India adopted directly observed treatment - short course (DOTS) under the Revised National Tuberculosis Control Programme (RNTCP) in 1992 and public-private mix DOTS in 2002. This study was conducted to assess the knowledge of doctors in the public and private sectors regarding TB control and management. STUDY DESIGN: Cross-sectional study. METHODS: This study used a self-reported questionnaire based on the RNTCP technical and operational guidelines. One hundred and forty one doctors were recruited through census sampling; all were registered with the Chest Physicians Association and treating TB using allopathic medicine. The list of doctors was obtained from Lucknow District TB Office, which annually updates and manages the members list of the Chest Physicians Association. This study was conducted in Lucknow, India in February-March 2007. RESULTS: Of 141 doctors, 71% had specialized medical education for treating TB, 60% had received RNTCP training and 69% reported that they follow DOTS methodology for TB treatment. Fifty-six percent of doctors worked in the public sector and 44% worked in the private sector. Forty-nine percent of doctors working in the public sector and 53% working in the private sector correctly reported all TB symptoms as per the RNTCP guidelines. Sixty-six percent of doctors in the public sector and 39% in the private sector reported the correct technique for sputum sampling. Public sector doctors demonstrated better knowledge of drug regimens for sputum smear positive and sputum smear-negative TB than private sector doctors. Statistical analysis indicated that doctors in the public sector had 2.1 times better knowledge than private sector doctors (odds ratio 2.1; P=0.05). CONCLUSION: Health policy managers and DOTS implementers should encourage all doctors, particularly private sector doctors, to receive RNTCP training and follow DOTS methodology. Improvement is needed in RNTCP training, and emphasis needs to be given to correct diagnosis, management and follow-up of TB patients. PMID- 19560179 TI - [Review of the neuroradiological diagnosis of cerebral venous thrombosis]. AB - Cerebral venous thrombosis is an uncommon entity with multiple causes. It must be diagnosed quickly to enable early treatment, avoid severe complications, and improve the patient's prognosis. MRI combined with MR venography is the best diagnostic approach. However, the clinical presentation is usually encountered in the emergency department, so cranial CT combined with CT venography is often the best way to establish the diagnosis because it has high sensitivity for the identification of the cerebral veins and sinuses. Although color-coded transcranial duplex sonography does not play a role in the initial diagnostic work-up, it is useful in the follow-up. We describe the distribution of the territories of venous drainage, analyze the usefulness of the different neuroradiological modalities, analyze the direct and indirect signs of central venous thrombosis, and discuss strategies to avoid the diagnostic pitfalls in patients with clinical suspicion of venous thrombosis. PMID- 19560180 TI - A new inorganic-organic composite coagulant, consisting of polyferric sulphate (PFS) and polyacrylamide (PAA). AB - Currently, research is focused on the synthesis of new composite coagulants, which are constituted of both inorganic and organic materials. In this paper, the development of relevant reagents was investigated, by combining the inorganic pre polymerised iron-based coagulant Polyferric Sulphate (PFS) with an organic, non ionic polymer (Polyacrylamide, PAA) under different PAA/Fe (mg/l) and OH/Fe molar ratios. Moreover, the new reagents were characterised in terms of typical properties, stability and morphological analysis (XRD, FTIR, SEM). Their coagulation performance, when treating low or high turbid kaolin-humic acid suspensions, was also investigated, whereas the applied coagulation mechanisms were discussed by using the Photometric Dispersion Analysis (PDA) analysis. The results show that the new coagulation reagents present improved properties, including increased effective polymer species concentration, and they exhibit very good stability. The respective tests using PDA confirmed that the predominant coagulation mechanism of PFS-PAA is the bridge formation mechanism. Coagulation experiments in low or high turbid kaolin-humic acid suspensions reveal that the novel composite reagent PFS-PAA exhibits better coagulation performance, when compared with simple PFS, in terms of zeta-potential reduction, turbidity and organic matter removal and residual iron concentration. PMID- 19560181 TI - Effect of pesticide concentration on the degradation process by combined solar photo-Fenton and biological treatment. AB - The influence of pesticide concentration, expressed as dissolved organic carbon (DOC), on combined solar photo-Fenton and biological oxidation treatment was studied using wastewater containing a mixture of five commercial pesticides, Vydate, Metomur, Couraze, Ditumur and Scala. Two initial DOC concentrations, 200 mg L(-1) and 500 mg L(-1) were assayed. Variation in biodegradability with photocatalytic treatment intensity was tested using Pseudomonas putida. Thus the mineralisation required for combining with biodegradation of intermediates by activated sludge was 33% and 55% at 200 mg L(-1) and 500 mg L(-1), respectively. Biotreatment was carried out in a stirred tank in sequencing batch reactor (SBR) mode. As revealed by the biodegradation kinetics, intermediates generated at the higher pesticide concentration caused lower carbon removal rates in spite of the longer photo-Fenton treatment time applied. One strategy for treating water with high concentrations of pesticides and overcoming the low biodegradability of photo-Fenton intermediates is to mix it with a biodegradable carbon source before biological oxidation. This combination of photo-Fenton and acclimatized activated sludge in several SBR cycles led to complete biodegradation of a concentrated pesticide solution of 500 mg L(-1) DOC in approximately 5h with a carbon removal efficiency of 90%. PMID- 19560183 TI - Enrichment of a K-strategist microbial population able to biodegrade p nitrophenol in a sequencing batch reactor. AB - The biological treatment of a high-strength p-nitrophenol (PNP) wastewater in an aerobic Sequencing Batch Reactor (SBR) has been studied. A specific operational strategy was applied with the main aim of developing a K-strategist PNP-degrading activated sludge. The enrichment of a K-strategist microbial population was performed using a non-acclimated biomass coming from a municipal WWTP as inoculum, and following a feeding strategy in which the PNP-degrading biomass was under endogenous conditions during more than 50% of the aerobic reaction phase. Hundred per cent of PNP removal was achieved in the whole operating period with a maximum specific PNP loading rate of 0.26 g PNP g(-1)VSS d(-1). A kinetic characterization of the obtained PNP-degrading population was carried out using respirometry assays in specifically designed batch tests. With the experimental data obtained a kinetic model including substrate inhibition has been used to describe the time-course of the PNP concentration and specific oxygen uptake rate (SOUR), simultaneously. The kinetic parameters obtained through optimization, validated with an additional respirometric test, were k(max)=1.02 mg PNP mg(-1) COD d(-1), K(s)=1.6 mg PNP L(-1) and K(i)=54 mg PNP L(-1). The values obtained for the K(s) and k(max) are lower than those reported in the literature for mixed populations, meaning that the biomass is a K-strategist type, and therefore demonstrating the success of the operational strategy imposed to obtain such a K strategist population. Moreover, our measured K(i) value is higher than those reported by most of the bibliographic references; therefore the acclimated activated sludge used in this work was evidently more adapted to PNP inhibition than the other reported cultures. PMID- 19560182 TI - Development of a TiO2 modified optical fiber electrode and its incorporation into a photoelectrochemical reactor for wastewater treatment. AB - Electrochemical advanced oxidation processes (EAOPs) are used to chemically burn non biodegradable complex organic compounds that are present in polluted effluents. A common approach involves the use of TiO2 semiconductor substrates as either photocatalytic or photoelectrocatalytic materials in reactors that produce a powerful oxidant (hydroxyl radical) that reacts with pollutant species. In this context, the purpose of this work is to develop a new TiO2 based photoanode using an optic fiber support. The novel arrangement of a TiO2 layer positioned on top of a surface modified optical fiber substrate, allowed the construction of a photoelectrochemical reactor that works on the basis of an internally illuminated approach. In this way, a semi-conductive optical fiber modified surface was prepared using 30 microm thickness SnO2:Sb films on which the photoactive TiO2 layer was electrophoretically deposited. UV light transmission experiments were conducted to evaluate the transmittance along the optical fiber covered with SnO2:Sb and TiO2 showing that 43% of UV light reached the optical fiber tip. With different illumination configurations (external or internal), it was possible to get an increase in the amount of photo-generated H(2)O(2) close to 50% as compared to different types of TiO2 films. Finally, the electro-Fenton photoelectrocatalytic Oxidation process studied in this work was able to achieve total color removal of Azo orange II dye (15 mg L(-1)) and a 57% removal of total organic carbon (TOC) within 60 min of degradation time. PMID- 19560184 TI - Occurrence of filamentous fungi and yeasts in three different drinking water sources. AB - In order to determine the occurrence of fungi in different drinking water sources and capture variability in terms of matrix composition and seasonal effects, surface water, spring water, and groundwater samples were collected in numerous sampling events. The occurrence and significance of fungi detected in the different water sources are reported and discussed in terms of colony-forming units per millilitre and by the identification of the most frequently detected isolates, at the species level, based on morphology and other phenotypic characters. All the samples were also analyzed in terms of total coliforms and Escherichia coli that are widely monitored bacteria considered as microbiology indicators of water quality. All the groundwater samples showed significantly lower levels of total coliforms, E. coli, and fungi compared to the surface and spring water samples. No significant correlations were found between the levels of fungi detected in all the matrices and the physico-chemical parameters and bacteria regularly monitored by drinking water utilities. Fifty-two fungi isolates were identified in this study, most of which have never been described to occur in water sources. The results obtained show that fungi occur widely in drinking water sources and that further studies should be conducted to address their biodegradation potential as well as if the drinking water treatment processes currently used are effective in removing these organisms and the potential secondary metabolites produced. PMID- 19560185 TI - Detecting points as developmental delay based on the life-history development and urosome deformity of the harpacticoid copepod, Tigriopus japonicus sensu lato, following exposure to benzo(a)pyrene. AB - To identify ecotoxicological responses to an endocrine disrupter, benzo(a)pyrene, we examined the life-history of the harpacticoid copepod, Tigriopus japonicus sensu lato. Based on the life-history of copepods, survival rate of nauplii (NSR) and copepodites (CSR), copepodite emergence day (CED) and adult male emergence day (AMED), sex ratio (MER), brooding success rate (BSR), and first brooding day of adult females (FBD) were measured. Significant differences were observed in the survival and development of nauplii (NSR and CED) and sex ratio (MER) of exposed and non-exposed copepods. Moreover, high concentration of BaP can be lethal to copepodite and exhibited a delay of growth. In this study, the CED and AMED among ecotoxicological response based on life-history developments were delayed and the body characteristics decreased in response to exposure to benzo(a)pyrene. The dwarfism and urosome deformity of the T. japonicus s.l. was exhibited in response to chemical exposure. Specifically, the body characteristics and biomass of dwarf copepods that had been exposed to benzo(a)pyrene were 30% and 50% lower than the control group, respectively. The incidence of abnormal urosomes was divided into two types. The first deformity type was signs of shrinkage in the middle of the urosome or the entire urosome was narrower than those of the control organisms. In the second type, the anal somite and the distal side of the urosome had abnormally swelled. Taken together, the nauplii and copepodid development of T. japonicus s.l. can be used as a useful biomaker for detecting developmental delay based on their entire life history. In addition, the urosome deformity was used a good potential monitoring tool invading various chemicals and environmental contamination into water system. PMID- 19560186 TI - Relative bioavailability of soil-bound polycyclic aromatic hydrocarbons in goats. AB - This study aimed at determining the relative bioavailability (RB) of three soil bound PAH model compounds (phenanthrene [PHE], pyrene [PYR] and benzo[a]pyrene [BaP]) in four lactating goats. RB was estimated by comparing the urinary or milk excretion of the major mono-hydroxylated metabolites of PAHs after ingestion of PAH spiked-soil and -oil feeds. A series of three increasing doses were orally administered in order to estimate the dose response of the two different matrices. The results of this study reveal that urinary excretion prevailed compared to milk excretion (30-fold higher). The recovery rate of mono hydroxylated metabolites of PAHs in urine and milk indicate that PYR was absorbed at a minimum level of 36%. 3-OH PHE excreted in urine suggests a minimal absorption of at least 5% for PHE. 3-OH BaP remained under the limits of detection and quantification and no RB could be calculated for this compound. RB of soil-bound PYR compared to PYR in oil was 61% and 50% in milk and urine, respectively. Thus, a significantly reduced RB of PYR in soil has been shown. On the other hand, no significant differences were observed between oil and soil for urinary 3-OH PHE (RB=100%). These results show that the soil matrix significantly reduces the bioavailability of certain PAHs. The decrease of bioavailability seems to be dependent on the compounds, i.e. higher for PYR than for PHE. This study also suggests that soil ingestion should be taken into account in risk assessment studies. PMID- 19560187 TI - Prothrombotic factors and the risk of acute onset non-cardioembolic stroke in young Asian Indians. AB - INTRODUCTION: Several prothrombotic factors--both hereditary and acquired--are known to cause stroke. Commonly investigated causes are activated protein C resistance, factor V Leiden mutation, factor VIII levels, prothrombin 20210 G-to A mutation, coagulation inhibitors such as proteins C and S, and antiphospholipid antibodies such as beta(2)-glycoprotein. OBJECTIVE: The literature on the prevalence of hematological defects pertaining to these variables in the Asian Indian stroke population is limited to a few isolated reports. In the current study we investigate the above-mentioned variables in 120 stroke patients (non cardioembolic acute-onset stroke) and compare their status with the hematological profile of an equal number of healthy age- and sex-matched controls. MATERIAL AND METHODS: Plasma and blood leukocytes were collected from all patients and controls for performing hematological assays and molecular tests respectively. The mutations were detected using standard polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) procedures. Statistical analysis was done using SPSS version 12.0. RESULTS: Factor V Leiden (prevalence 8.3% in patients) and activated protein C resistance (prevalence 19.6% in patients) both showed a high degree of association (P<0.01) with the disease condition. However, contrary to common expectations, factor V Leiden was observed much less frequently in patients showing activated protein C resistance (10 out of 23; 43.4%) than is commonly observed in the Caucasian population (almost 90%). Post-acute-phase factor VIII levels were also found to be significantly associated with stroke: 125.6+21.1% number of profitable positions (NPP) for controls and 136.2+28.8% NPP for patients (P=0.001). CONCLUSION: factor V mutations, such as factor V Leiden, may be important risk factors for stroke in an Asian Indian population. Activated protein C resistance has a stronger association with stroke than factor V Leiden and may be caused by other factors such as elevated factor VIII levels in the Asian Indian population apart from factor V Leiden itself. PMID- 19560188 TI - Follow-up for women after treatment for cervical cancer: a systematic review. AB - OBJECTIVE: To determine the optimal recommended program for the follow-up of patients who are disease free after completed primary therapy for cervical cancer. METHODS: Systematic search of MEDLINE, EMBASE and the Cochrane Library databases (1980-November 2007). RESULTS: Seventeen retrospective trials were identified. Most studies reported similar intervals for follow-up and ranged from a low of 9 visits to a high of 28 visits over 5 years. Follow-up visits typically occurred once every 3-4 months for the first 2 years, every 6 months for the next 3 years and then annually until year 10. All 17 trials reported that a physical exam was performed at each visit. Vaginal vault cytology was analyzed in 13 trials. Other routine surveillance tests included chest x-ray, ultrasound, CT scans, MRI, intravenous pyelography and tumour markers. Median time to recurrence ranged from 7-36 months after primary treatment. Rates of recurrence ranged from 8-26% with 14-57% of patients recurring in the pelvis, and 15-61% of patients recurring at distant or multiple sites. Of the 8-26% of patients who experienced disease recurrence, the vast majority, 89-99%, had recurred by year 5. Upon recurrence, median survival was 7-17 months. Asymptomatic recurrent disease was detected using physical exam in 29-71%, chest x-ray in 20-47%, CT in 0-34% and vaginal vault cytology in 0-17% of patients, respectively. CONCLUSION: There is modest low quality evidence to inform the most appropriate follow-up strategy for patients with cervical cancer who are clinically disease free after receiving primary treatment. Follow-up visits should include a complete physical examination whereas, frequent vaginal vault cytology does not add significantly to the detection of early disease recurrence. Patients should return to annual population-based screening after 5 years of recurrence-free follow-up. PMID- 19560189 TI - Outcomes associated with different intraperitoneal chemotherapy delivery systems in advanced ovarian carcinoma: a single institution's experience. AB - BACKGROUND: Despite increasing use of intraperitoneal chemotherapy the optimal delivery strategy and regimen remain undetermined. Catheter-related complications have been reported in 3-34% of cases across a number of platforms and port styles, but few data compare different catheters directly. We sought to evaluate the complication rate of two separate intraperitoneal chemotherapy port delivery systems used within a single practice. METHODS: We reviewed the medical records of all patients who underwent port placement in our practice (two surgical centers) from January, 2006 through October, 2008. Data extracted included: demographics, medical co-morbidities, port type, timing of placement, intraoperative procedures, reasons for discontinuation of IP chemotherapy, and number of completed cycles. RESULTS: We identified 85 patients who had intraperitoneal ports placed. Four patients were excluded from this analysis: 2 declined chemotherapy and 2 were treated at other institutions and follow-up data was insufficient. Fifty-two (64%) of the 81 patients analyzed had a fenestrated port placed, and 29 (36%) had single lumen ports. In 67 cases (83%) the port was placed at the time of initial cytoreductive surgery. In 14 patients (17%) it was placed as a secondary event. The groups were well matched for age, stage, BMI, and medical co-morbidities though the group with single lumen catheters had more antecedent surgeries. We observed no significant difference between patients with single lumen or fenestrated ports with regard to: number of intraperitoneal treatments, catheter-related complications, hematologic outcomes, and rates of discontinuation. CONCLUSIONS: A low rate of catheter-related complications is observed with both systems. The majority of discontinuations were due to hematologic complications and did not appear to be intrinsic to catheter choice. PMID- 19560190 TI - A case of multi-modal managed vulval aggressive angiomyxoma diagnosed before conception and monitored during pregnancy. PMID- 19560191 TI - Racial disparities in survival among patients with germ cell tumors of the ovary- United States. AB - OBJECTIVE(S): To compare the survival of African American (AA) and white (W) patients with malignant germ cell tumors of the ovary (OGCT). METHODS: Patients with a diagnosis of OGCT were identified from Surveillance, Epidemiology, and End Results Program (SEER) from 1988 to 2004, and were divided into African American (AA) and white (W) subgroups. Only surgically treated patients were included. Histology was grouped into dysgerminoma (D), malignant teratoma (MT), and mixed germ cell tumors with pure non-dysgerminoma cell tumors (MGCT/PNDCT). Statistical analysis using Chi-square, Fisher's Exact Test, Kaplan-Meier survival methods, and Cox regression proportional hazards were performed. RESULTS: In 1110 patients with OGCT, 970 (87.4%) were W and 140 (12.6%) were AA. MGCT/PNDCT histology was equally represented in AA and W. However, W were twice as likely to present with D (W 34% vs. AA 16%, p<0.01) and 1.5 times less likely to present with MT (W 41% vs. AA 59%, p<0.01). The majority (W 64%, AA 64%) of OGCT were stage I. Advanced stage (FIGO III and IV) tumors were more prominent in AA (24% vs. 18%, p>0.05). Complete surgical staging effort was utilized more frequently in W (49%) as compared to AA (38%; p=0.001). Overall 5-year survival was 92% for W and 86% for AA (p=0.02). In multivariate analysis race was not an independent predictor of survival when histology, stage and surgical staging were controlled. CONCLUSION(S): In our study, a higher prevalence of complete surgical staging and a favorable distribution of low risk histologic types may explain the improved survival observed in white patients with OGCT. However, race was not an independent predictor of survival. PMID- 19560192 TI - Estrogen and progesterone receptor status and outcome in epithelial ovarian cancers and low malignant potential tumors. AB - OBJECTIVE: Receptors for estrogen (ER) and progesterone (PR) are prognostic indicators for a variety of endocrine tumors including breast and endometrial. This study was conducted to determine if ER and PR expression patterns are predictive of outcome in patients with epithelial ovarian cancer (EOC) or ovarian low malignant potential (LMP) tumors. METHODS: ER and PR protein levels were assessed by immunohistochemistry in 45 LMP and 89 EOC samples. Patterns of ER/PR expression (individually and combinations of ER-/PR-, ER+/PR-, ER-/PR+, and ER+/PR+) were correlated with standard prognostic factors of overall survival (OS) in this patient population. RESULTS: For patients with EOC, the 5-year OS per ER-/PR+, ER+/PR-, ER+/PR+, and ER-/PR- expression was 83%, 79%, 61%, and 48%, respectively, and these differences were statistically significant. In multivariate analyses, ER/PR expression patterns were found to be independent predictors of OS, as were the classical prognostic factors of grade, stage, debulking, and chemotherapy response to treatment. In patients with mucinous LMP tumors, ER and PR were absent. Because no LMP patients died of disease during the studied period, no correlation analysis with OS could be performed. CONCLUSIONS: Patterns of ER/PR expression provide prognostic information in EOC. Additional studies evaluating hormonal inhibition may help personalize the therapy of patients with ovarian cancer. PMID- 19560193 TI - A multicenter evaluation of sequential multimodality therapy and clinical outcome for the treatment of advanced endometrial cancer. AB - OBJECTIVES: The appropriate sequencing of chemotherapy and radiation for the treatment of advanced endometrial cancer has not yet been determined. We sought to evaluate the outcome and adverse effects in patients with advanced stage endometrial cancer treated with postoperative chemotherapy and radiation to determine whether there was an advantage to a particular sequencing modality. METHODS: A multicenter retrospective analysis of patients with surgical stages III and IV endometrial cancer from 1993 to 2007 was conducted. Inclusion criteria were comprehensive staging procedure including hysterectomy, bilateral salpingo oophorectomy, +/- selective pelvic/aortic lymphadenectomy, and treatment with adjuvant chemotherapy and radiation. Differences in frequencies of adverse events were tested with Pearson's chi-square test for comparing proportions. OS and PFS rates were calculated using Kaplan-Meier estimates. Hazard Ratios (HR) were estimated from multivariate Cox proportional hazards models. RESULTS: One hundred and nine patients with advanced stage endometrial cancer were identified who received postoperative adjuvant therapies; 41% (n=45) chemotherapy followed by radiation and then further chemotherapy (CRC), 17% (n=18) radiation followed by chemotherapy (RC), and 42% (n=46) chemotherapy followed by radiation (CR). The median age was 62 years (range: 35-83); 48% had endometrioid tumors; and 90% underwent optimal cytoreduction. There was no difference in the frequency of adverse effects due to either chemotherapy (p=0.35) or radiotherapy (p=0.14); dose modifications (p=0.055); or delays (p=0.80) between the various sequencing modalities. There was a significant difference between adjuvant treatment groups for both OS (log rank p=0.011) and PFS (log rank p=0.025), with those receiving CRC having a superior 3-year OS (88%) and PFS (69%) compared to RC (54% and 47%) or CR (57% and 52%). After adjusting for stage, age, grade, race, histology and cytoreduction status the OS HR for therapy was 5.74 (95% CI, 1.96 to 16.77) for RC and 2.60 (95% CI, 1.01 to 6.71) for CR, compared to CRC, p=0.003. When the analysis was restricted to optimally cytoreduced patients, those who were treated with RC were at higher risk for disease progression [HR=3.53 (95% CI, 1.29 to 9.71)], p=0.024, and death [HR=7.24 (95% CI, 2.25 to 23.37)], p=0.001, than patients who received sequential CRC. CONCLUSIONS: Sequential CRC was associated with improved survival in women with advanced stage disease compared to other sequencing modalities with a similar adverse effect profile. Future clinical trials are needed to prospectively evaluate appropriate sequencing and types of adjuvant chemotherapy and radiotherapy for the treatment of advanced stage endometrial cancer. PMID- 19560194 TI - Prevalence of obstructive sleep apnea-hypopnea in severe versus moderate asthma. AB - BACKGROUND: Previous studies have suggested a link between obstructive sleep apnea and poor asthma control, which may be mediated through airway inflammation, obesity, and other mechanisms. OBJECTIVE: To test the hypothesis that the prevalence and severity of sleep apnea is greater among patients with severe compared with moderate asthma and controls without asthma. METHODS: Complete overnight home polysomnography was performed in 26 patients with severe asthma consecutively recruited to a difficult asthma program, 26 patients with moderate asthma, and 26 controls without asthma of similar age and body mass index. Flow rates and Juniper asthma control and quality of life questionnaires were also obtained. RESULTS: Obstructive sleep apnea-hypopnea, defined by an Apnea-Hypopnea Index > or = 15 events/h of sleep scored using Chicago criteria, was present in 23 of 26 (88%) patients with severe asthma, 15 of 26 (58%) patients with moderate asthma, and 8 of 26 (31%) controls without asthma (chi(2): P < .001). Using the more restrictive scoring criteria applied in the Wisconsin cohort study, Apnea Hypopnea Index > or = 5/h was present in 50% (severe), 23% (moderate), and 12% (control) of subjects (P = .007). Mean nocturnal arterial oxygen saturation was significantly lower in patients with severe asthma versus controls, and apnea hypopnea severity measures were significantly worse for both asthmatic groups compared with controls. Among subjects with asthma, no significant correlations were identified between the severity of sleep-disordered breathing and asthma severity or control measures (FEV(1), Juniper scores). CONCLUSIONS: Obstructive sleep apnea-hypopnea was significantly more prevalent among patients with severe compared with moderate asthma, and more prevalent for both asthma groups than controls without asthma. These observations suggest potential pathophysiologic interactions between obstructive sleep apnea-hypopnea and asthma severity and control. PMID- 19560195 TI - Effect of composition, viscosity and thickness of the opaquer on the adhesion of resin composite to titanium. AB - OBJECTIVES: The objectives of this study were to determine the bond strength of powder-liquid and paste opaquers with different chemical compositions and viscosity to a metal substructure when they were applied in two thicknesses and to evaluate the failure modes after the bond strength test. METHODS: Titanium plates (51 mm x 25 mm x 1mm) (n(plates)=25, N=80, n=10 per group) were conditioned with chairside silica coating (CoJet-Sand, 30 microm silica coated Al(2)O(3)) from a distance of approximately 10mm at a pressure of 2.8 bar for 15 s/cm(2) and silanized. Four types of opaquers, namely one powder-liquid (Sinfony, 3M ESPE), and three paste opaquers [(Cimara, Voco), (Monopaque, Ivoclar Vivadent), (Cavex Experimental, Cavex)] were applied either in 0.25 or 0.50 mm thicknesses using standard polyethylene molds and photo-polymerized. Resin composite (Quadrant Posterior Dense, Cavex) was applied incrementally and photo polymerized. The specimens were thermocycled (5-55 degrees C, 6000 cycles) prior to shear bond strength test (1mm/min). Failure types were analyzed using an optical microscope and scores were given according to the modified Adhesive Remnant Index (ARI) (Score 0=no opaquer on the surface, Score 1=<1/2 covered with opaquer, Score 2=>1/2 covered with opaquer, Score 3=completely covered with opaquer). RESULTS: While thickness did not significantly affect the bond strength results (p=0.523), type of opaquers had a significant influence on the results (p<0.01) (Univariate ANOVA, Tukey's test). Interaction terms between thickness and opaquer type were significant (p<0.01). Debonded specimens during thermocycling were considered as 0 MPa. At both 0.25 and 0.5mm thicknesses, powder-liquid based opaquer (Sinfony) showed significantly higher results (8.4+/ 5.6 and 8.4+/-4.9 MPa, respectively) than those of other opaquers (1.4+/-1 to 4.3+/-3.8 MPa) (p<0.05). Only when Cimara was applied in 0.25 mm (6.9+/-4.2 MPa), there were no significant differences with Sinfony (p>0.05). The lowest results in both thicknesses were obtained from Monopaque (4+/-3.8 to 1.6+/-1 MPa, respectively) and Cavex (1.4+/-1 to 4.2+/-2.9 MPa, respectively) paste opaquers. In all opaquers, the incidence of Score 0 (30) was more frequent followed by Score 1 (27) and Score 2 (20). SIGNIFICANCE: The use of powder-liquid opaquer in order to mask the metal in repair actions provided higher bond strength than those of the paste opaquers in both thin and thick applications. In all opaquers, the incidence of adhesive failure between the opaquer and the metal was more common implying inadequate adhesion. PMID- 19560196 TI - The short version of the Depression Anxiety Stress Scales (DASS-21): factor structure in a young adolescent sample. AB - This study explored the factor structure of the short form of the Depression Anxiety Stress Scales (DASS-21; Lovibond & Lovibond, 1995b) in a young adolescent sample. A group of 484 high school students (Mean age=13.62 years, Min=11.83, Max=15.67 years, 52 % boys) completed the DASS-21. Several models were tested using Confirmatory Factor Analysis. A model consistent with the factor structure of the adult DASS, with correlated error terms, provided good fit to the data. However, correlations among the factors were very high. A 'quadripartite' model involving a common 'Negative Affect' factor as well as the three specific factors of Depression, Anxiety and Tension/Stress was tested to explain these correlations and was supported by the data. This model suggests that the core symptoms of Depression and Anxiety are similar in adults and adolescents, but the conceptualisation and assessment of Tension/Stress in adolescents needs further refinement. PMID- 19560197 TI - Hemoglobin conjugated micelles based on triblock biodegradable polymers as artificial oxygen carriers. AB - An artificial oxygen carrier is constructed by conjugating hemoglobin molecules to biodegradable micelles. Firstly a series of triblock copolymers (PEG-PMPC-PLA) in which the middle block contains pendant propargyl groups were synthesized and characterized. After the amphiphilic copolymer was self-assembled into core-shell micelles in aqueous solution, azidized hemoglobin molecules protected by carbon monoxide (CO) were conjugated to the micelles via click reaction between the propargyl and azido groups. The conjugation causes an increase of the micelle's mean diameter. Maximum conjugation ratio is 250 wt% in the hemoglobin-conjugated micelles (HCMs). Oxygen-binding ability of the HCMs was demonstrated by converting the CO-binding state of the HCMs into O(2)-binding state. PMID- 19560198 TI - Protein adsorption and cell adhesion on cationic, neutral, and anionic 2 methacryloyloxyethyl phosphorylcholine copolymer surfaces. AB - Protein adsorption and cell adhesion on cationic, neutral, and anionic water soluble 2-methacryloyloxyethyl phosphorylcholine (MPC) copolymer surfaces were compared. These model MPC copolymers coated SiO(2) surfaces exhibited comparable surface zeta-potentials of 26.1 mV, near 0 mV, and -24.2 mV, respectively. X-ray photoelectron spectroscopy analyses indicated the similarities and the differences in the surface composition between the sample surfaces. Atomic force microscopy analyses revealed that the type of the charged moiety did not affect the surface roughness. Static contact angle measurements and dynamic contact angle analyses not only indicated that the surfaces were very hydrophilic in general, but also provided information on the surface mobility and the dominant role of MPC at the surface in aqueous conditions. Comparing with the SiO(2) substrates on which protein seriously adsorbed and cell heavily adhered, three MPC copolymers coated surfaces, despite their different charge properties, exhibited significantly low adsorbed amounts of different proteins having various electrical natures and totally no cell adhesion. This suggested that the incorporation of charged moieties in the MPC copolymers did not significantly inspire both the protein adsorption and cell adhesion. The MPC moieties were predominant at the surface when in contact with aqueous conditions and thereby dominated the bio-adsorptions, while the possible effect from electrostatic interactions would be too small and too limited to influence the overall situation. Therefore, these MPC copolymer surfaces can satisfy those biological applications requiring not only electrical but also non-biofouling properties. PMID- 19560199 TI - Folic acid-Pluronic F127 magnetic nanoparticle clusters for combined targeting, diagnosis, and therapy applications. AB - Superparamagnetic iron oxides possess specific magnetic properties in the presence of an external magnetic field, which make them an attractive platform as contrast agents for magnetic resonance imaging (MRI) and as carriers for drug delivery. In this study, we investigate the drug delivery and the MRI properties of folate-mediated water-soluble iron oxide incorporated into micelles. Pluronic F127 (PF127), which can be self-assembled into micelles upon increasing concentration or raising temperatures, is used to decorate water-soluble polyacrylic acid-bound iron oxides (PAAIO) via a chemical reaction. Next, the hydrophobic dye Nile red is encapsulated into the hydrophobic poly(propylene oxide) compartment of PF127 as a model drug and as a fluorescent agent. Upon encapsulation, PAAIO retains its superparamagnetic characteristics, and thus can be used for MR imaging. A tumor-specific targeting ligand, folic acid (FA), is conjugated onto PF127-PAAIO to produce a multifunctional superparamagnetic iron oxide, FA-PF127-PAAIO. FA-PF127-PAAIO can be simultaneously applied as a diagnostic and therapeutic agent that specifically targets cancer cells that overexpress folate receptors in their cell membranes. PF127-PAAIO is used as a reference group. Based on FTIR and UV-vis absorbance spectra, the successful synthesis of PF127-PAAIO and FA-PF127-PAAIO is realized. The magnetic nanoparticle clusters of PF127-PAAIO and FA-PF127-PAAIO are visualized by transmission electron microscope (TEM). FA-PF127-PAAIO, together with a targeting ligand, displays a higher intracellular uptake into KB cells. This result is confirmed by laser confocal scanning microscopy (CLSM), flow cytometry, and atomic absorption spectroscopy (AAS) studies. The hysteresis curves, generated by using a superconducting quantum interference device (SQUID) magnetometer analysis, demonstrate that the magnetic nanoparticles are superparamagnetic with insignificant hysteresis. The MTT assay explains the negligible cell cytotoxicity of PF127-PAAIO and FA-PF127-PAAIO. In KB cells, the in vitro MRI study indicates the better T(2)-weighted images in FA-PF127-PAAIO than in PF127-PAAIO. PMID- 19560200 TI - Whole proteome analysis of osteoprogenitor differentiation induced by disordered nanotopography and mediated by ERK signalling. AB - Topographic features can modulate cell behaviours such as proliferation, migration, differentiation and apoptosis. Biochemical mechanotransduction implies the conversion of mechanical forces (e.g. changes in cell spreading and morphology from changing surface topography) into biochemical signal via biomolecules. Still, little is known concerning which pathways may be directly involved in cell response to changes in the material surface. A number of pathways have been implicated using focused studies of 'selected' biomolecules rather than a global analysis of signal pathways. This study used a controlled disorder nanopit topography (NSQ50, fabricated by electron beam lithography) to direct osteoblast differentiation of progenitor cells. This topography is unique as it represents a middle route (from absolute order or random roughness) that allows osteoconversion with similar efficiency as dexamethasone and ascorbate treatment. Two direct-comparison proteomics techniques, firstly gel-based and then chromatography-based, were used to analyse progenitor proteome changes in response to the nanotopography. Many of the changed proteins form part of the Extracellular Signal-regulated Kinase (ERK1/2) pathway. PMID- 19560201 TI - Human decidual stromal cells protect lymphocytes from apoptosis. AB - Human decidual stromal cells (DSC) have been shown to be involved in different immune functions that may be relevant for the relationship between the mother and fetus and hence for successful pregnancy. The expression of death ligands by fetal trophoblast and maternal decidual cells has been proposed as a mechanism for the establishment of materno-fetal immunotolerance. This study intended to elucidate the interrelations between DSC and lymphocytes. We analyzed the expression and function of death receptors and ligands in DSC maintained in culture. These DSC lines expressed CD95 and TNF-related apoptosis-inducing ligand receptor-2 (TRAIL-R2), although they were resistant to death receptor-mediated apoptosis. Regarding the expression of CD95L and TRAIL, it was variable among DSC lines although none of them induced apoptosis in death ligand-sensitive Jurkat T cells. Interestingly, most of the DSC lines, as well as fresh DSC, reduced apoptosis in Jurkat cells induced by anti-CD95 antibody and recombinant TRAIL. The protective effect of DSC was observed when they were co-cultured with Jurkat cells in Transwell plates, indicating that DSC may produce soluble factors of importance for lymphocyte survival. Moreover, the viability of peripheral blood lymphocytes and decidual lymphocytes was improved when co-cultured with DSC. Our results suggest that DSC, far from inducing apoptosis, may be relevant in the regulation of lymphocyte survival at the materno-fetal interface. PMID- 19560202 TI - Does oxidative damage contribute to the generation of leukemia? PMID- 19560203 TI - An environmental risk assessment for oseltamivir (Tamiflu) for sewage works and surface waters under seasonal-influenza- and pandemic-use conditions. AB - In the event of an influenza pandemic, anti-viral medications such as oseltamivir (Tamiflu) are expected to be used in high amounts over a duration of several weeks. Oseltamivir has been predicted to reach high concentrations in surface waters and sewage works. New oseltamivir environmental fate and toxicity studies permit an environmental risk assessment (ERA) under seasonal- and pandemic-use scenarios. The environmental fate data for sewage works (no removal), surface waters (no significant degradation), and water/sediment systems (>50% primary degradation in 100 days) were used for the derivation of new predicted environmental concentrations (PECs) for western Europe and the River Lee catchment in the UK. Existing worst-case PECs for western Europe, the River Lee catchment in the UK and the Lower Colorado basin in the USA under pandemic conditions (< or =98.1 microg/L for surface waters, < or =348 microg/L for sewage works) were also considered for the ERA. PECs were compared with predicted no effect concentrations (PNECs) based on new chronic ecotoxicity data (no observed effect concentration for algae, daphnia, and fish > or =1 mg/L). Based on all PEC/PNEC risk ratios, no significant risk is evident to surface waters or sewage works during both regular seasonal-use and high pandemic-use of oseltamivir. PMID- 19560204 TI - Genomic organization and evolution of immunoglobulin kappa gene enhancers and kappa deleting element in mammals. AB - We have studied the genomic structure and evolutionary pattern of immunoglobulin kappa deleting element (KDE) and three kappa enhancers (KE5', KE3'P, and KE3'D) in eleven mammalian genomic sequences. Our results show that the relative positions and the genomic organization of the KDE and the kappa enhancers are conserved in all mammals studied and have not been affected by the local rearrangements in the immunoglobulin kappa (IGK) light chain locus over a long evolutionary time ( approximately 120 million years of mammalian evolution). Our observations suggest that the sequence motifs in these regulatory elements have been conserved by purifying selection to achieve proper regulation of the expression of the IGK light chain genes. The conservation of the three enhancers in all mammals indicates that these species may use similar mechanisms to regulate IGK gene expression. However, some activities of the IGK enhancers might have evolved in the eutherian lineage. The presence of the three IGK enhancers, KDE, and other recombining elements (REs) in all mammals (including platypus) suggest that these genomic elements were in place before the mammalian radiation. PMID- 19560206 TI - Intravitreal ranibizumab may induce retinal arteriolar vasoconstriction in patients with neovascular age-related macular degeneration. AB - OBJECTIVE: To study the effect of intravitreal (IVT) ranibizumab (Lucentis; Genentech, Inc, San Francisco, CA) on the retinal arteriolar diameter in patients with neovascular age-related macular degeneration (AMD). DESIGN: Prospective consecutive interventional case series. PARTICIPANTS: Eleven eyes of eleven patients with previously untreated neovascular AMD. METHODS: All eyes had 3 monthly IVT injections of ranibizumab. The diameter of the retinal arterioles was measured in vivo with a retinal vessel analyzer (RVA) before the first IVT injection and then 7 and 30 days after the first, second, and third injections. MAIN OUTCOME MEASURES: Primary end points were changes in retinal arteriolar diameter and mean arterial pressure (MAP) after IVT ranibizumab. Secondary end points were changes in best-corrected visual acuity (BCVA), central retinal thickness, and intraocular pressure after IVT ranibizumab, and appearance of adverse events during the follow-up period. RESULTS: A significant decrease of the retinal arteriolar diameter was observed after each IVT injection of ranibizumab. Thirty days after the first, second, and third injections, there was a mean decrease of 8.1+/-3.2%, 11.5+/-4.4%, and 17.6+/-7.4%, respectively, of the retinal arteriolar diameter compared with baseline values (P<0.01). There was no significant change in MAP during the period of follow-up (P>0.05). Thirty days after the third IVT injection of ranibizumab, mean BCVA improved by 6.5+/-4.9 Early Treatment Diabetic Retinopathy Study (ETDRS) letters, and central retinal thickness decreased by 91+/-122 microm (P = 0.03). CONCLUSIONS: These results suggest that IVT ranibizumab may induce retinal arteriolar vasoconstriction in patients with neovascular AMD after IVT ranibizumab. Further studies evaluating larger sample sizes are needed to confirm these results and potential adverse effects on the retinal circulation in patients with AMD and retinal vascular diseases. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article. PMID- 19560205 TI - Macular and nerve fiber layer thickness in amblyopia: the Sydney Childhood Eye Study. AB - PURPOSE: To examine macular and peripapillary retinal nerve fiber layer (RNFL) thickness in amblyopia. DESIGN: Population-based cross-sectional study. PARTICIPANTS: Of 4118 children examined in the Sydney Childhood Eye Study (incorporating the Sydney Myopia Study) from 34 randomly selected primary schools and 21 secondary schools from 2003 to 2005, 3529 (85.7%) were included in this analysis. The median age of the 2 samples was 6 years (n = 1395) and 12 years (n = 2134), respectively. METHODS: A detailed eye examination was conducted on all children, including determination of best-corrected visual acuity (logarithm of the minimum angle of resolution [logMAR]), autorefraction (RK-F1 autorefractor, Canon, Tokyo, Japan) after cyclopentolate (1%), cover testing to identify strabismus, and optical coherence tomography (StratusOCT, Carl Zeiss Meditec, Dublin, CA) through dilated pupils to obtain macula and peripapillary RNFL thickness. Amblyopia was defined as best visual acuity <0.3 logMAR units not explained by any obvious underlying eye or visual pathway abnormalities. Anisometropia was defined as an interocular difference of at least 1.0 diopter of the spherical equivalent refraction. MAIN OUTCOME MEASURES: Macular and peripapillary RNFL thickness. RESULTS: Amblyopic eyes had slightly greater foveal minimum thickness than the normal fellow eye (by 5.0 microm; 95% confidence interval 0.1-9.9) and right eyes of non-amblyopic children (by approximately 10 microm), both P<0.05. This was more pronounced in 6-year-old children (6.9 microm) than 12-year-old children (4.2 microm). Amblyopic eyes also had slightly thicker central macula (1 mm diameter region) in both comparisons, although these differences were not statistically significant. The inner macular ring (outer radius 1.5 mm) was thinner in amblyopic than normal fellow eyes. Peripapillary RNFL thickness was not significantly different between amblyopic and normal fellow eyes or normal eyes of non-amblyopic children. CONCLUSIONS: In children aged predominantly 6 and 12 years, central macular thickness may be increased in eyes with amblyopia, although it is uncertain if this precedes or follows the development of amblyopia. No differences in peripapillary RNFL thickness were found when compared with normal eyes. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article. PMID- 19560207 TI - Feasibility of saccadic vector optokinetic perimetry: a method of automated static perimetry for children using eye tracking. AB - PURPOSE: To determine the feasibility of a new technique for suprathreshold automated static perimetry in children. DESIGN: Evaluation of diagnostic test or technology. PARTICIPANTS: The study included 29 subjects comprising 4 groups: 12 adults with normal fields, 4 children aged less than 10 years with normal fields, 8 adults with visual field defect, and 5 children aged less than 10 years with suspected visual field defects. METHODS: The system comprises a personal computer, display, and eye tracker to monitor gaze position when stimuli are presented in the visual field. The natural saccadic eye movement to fixate on the stimuli, if seen, can be detected and measured to produce a visual field plot. Subjects performed 3 eye-tracking tests, unless unable to do so for any reason: a 40-point binocular test and two 41-point tests for each eye. The tests were based on the Humphrey Field Analyzer (HFA) Central-40 point screening test with a stimulus size of Goldmann III and intensity of 14 decibels (dB). Adults also performed the equivalent Humphrey screening test in each eye for comparison. MAIN OUTCOME MEASURES: Comparison of visual field plot results between the eye tracking tests and HFA tests in adults. Correlation between the eye-tracking tests and the clinical assessment in the children with suspected visual field defects. RESULTS: In the eyes of all normal adult and child subjects performing the eye-tracking test, the percentage of points in agreement with a healthy visual field was 99.2% and 99.1%, respectively. The percentage of points agreeing with the HFA's screening test in the adult eyes with visual field defects was 89.8%. Visual field defects were also correctly identified by the eye-tracking system in the eyes of children with suspected visual field defects. CONCLUSIONS: The results demonstrate that suprathreshold automated static perimetry using eye tracking is a promising method of perimetry for use with children. PMID- 19560208 TI - Visual function in working-age adults: early life influences and associations with health and social outcomes. AB - PURPOSE: To investigate how visual function in mid-adult life is associated with health and social outcomes and, using life-course epidemiology, whether it is influenced by early life biological and social factors. DESIGN: Population-based cohort study. PARTICIPANTS: Nine thousand three hundred thirty members of the 1958 British birth cohort at age 44 or 45 years. METHODS: Distance, near, and stereo vision were assessed as part of a broader biomedical examination. Logistic, multinomial, and proportional odds ordinal logistic regression were used, as appropriate, to assess the association between these vision functions and both key early life influences and health and social outcomes in mid-adult life. MAIN OUTCOME MEASURES: Distance, near, and stereo acuities and health and social outcomes. RESULTS: In mid-adult life, vision function (across the full spectrum of both type and level of function) is associated with unemployment resulting from permanent sickness, lower socioeconomic status, and poorer general health (for example, for blindness; odds ratios were 2.5, 2.6, and 1.2, respectively). Also, impaired visual functions in mid-adult life are associated with a low birthweight, being small for gestational age, maternal smoking in pregnancy, and markers of socioeconomic deprivation in childhood (for example, for impaired distance acuity; odds ratios were 1.4, 1.3, 1.02, and 1.1, respectively). CONCLUSIONS: Although relatively uncommon in working-age adults, impaired vision can have important adverse consequences, which highlights the value of investigating visual function in the broader context of health and social functioning. In addition, visual function in adult life may be influenced directly by key prenatal and childhood biological and social determinants of general health. Thus, application of life-course epidemiology to complex chronic ophthalmic diseases of adult life such as glaucoma or macular degeneration is likely to prove valuable in elucidating whether and how biological, social, and lifestyle factors contribute to the cause. PMID- 19560209 TI - Cytotoxicity of cis-platinum(II) cycloaliphatic amidine complexes: Ring size and solvent effects on the biological activity. AB - A series of new platinum(II) amidine derivatives of the type cis-[PtCl(2){Z NHC(NHR)Me}(2)] (R=cyclopropyl, 1; cyclopentyl, 2; cyclohexyl, 3) were prepared in high yield by addition of the corresponding cyclic aliphatic amine RNH(2) to the coordinated acetonitrile ligands in cis-[PtCl(2)(NCMe)(2)]. The solution behaviour of 1-3 has been studied in DMSO, PEG 400 (polyethylene glycol) and PEG DME 500 (polyethylene glycol dimethylether). The amidine complexes 1-3 were evaluated for their cytotoxic properties against a panel of human tumor cell lines containing examples of cervix (HeLa), breast (MCF7), lung (A549) and colon (HCT-15) cancer. Moreover, the amidine complexes were tested for their cytotoxicity against normal human fibroblasts (HFF-1). For comparison purposes, the cytotoxicity of cisplatin was examined under the same experimental conditions. The results obtained showed that PEG and PEG-DME behave as good solvents to carry out biological assays with platinum complexes which are water insoluble and unstable in DMSO. Complexes 2 and 3 exhibited a biological activity comparable to that of cisplatin. PMID- 19560211 TI - Immune response to Leishmania antigen in anthroponotic cutaneous leishmaniasis. AB - BACKGROUND: Leishmania (L.) tropica is the causative agent of anthroponotic cutaneous leishmaniasis (ACL) in Iran. The disease often heals within a year; however, the non-healing forms of disease are also known. The immunologic responses to L. major infection have been studied in depth, however little is known about the immune status of L. tropica-infected patients. MATERIALS AND METHODS: This study was conducted to evaluate T-cell responses to Leishmania antigen in non-healing patients, patients with acute lesion, and healthy donors. Peripheral blood mononuclear cells (PBMC) were cultured with antigen and lymphoproliferative responses were determined. Cytokine profile including gamma interferon (IFN-gamma), interleukin (IL)-5, and IL-13 in supernatants of stimulated cells was also determined. RESULTS: The results showed PBMC from both groups of patients proliferated vigorously in response to Leishmania antigens. The levels of IFN-gamma and IL-13 were comparable between patients with acute lesions and non-healing patients. Non-healing patients had significantly higher median levels of IL-5 than patients with acute lesions. The cells from healthy individuals did not respond to Leishmania antigens. CONCLUSIONS: High levels of IFN-gamma, IL-5, and IL-13 in non-healing patients suggest a mixed Th1/Th2 response, whereas patients with acute lesion respond to infection by Th1-type response. PMID- 19560210 TI - Severe invasive Panton-Valentine Leucocidin positive Staphylococcus aureus infections in children in London, UK. AB - OBJECTIVE: To describe the features of invasive Panton-Valentine Leucocidin positive Staphylococcus aureus (PVL-SA) in children at a London teaching hospital, from 2004 to 2008. METHODS: Retrospective case note review. RESULTS: Eleven previously healthy children, 7 male, median age 9 years (range 7 months-13 years), had invasive infections due to unrelated community-acquired meticillin sensitive PVL-SA. Possible risk factors were identified in 10 cases. Eight patients had complicated musculoskeletal infections, 2 had pneumonia, and 1 had a massive retropharyngeal abscess. At admission neutropenia was present in 2 patients, deep vein thrombosis in 3, and initial blood cultures were positive in 8. Patients with musculoskeletal involvement had a median of 3 (range 1-6) sites of infection, and required median 5 (range 1-11) operative procedures. Eight patients were admitted to PICU, 7 had septic shock. Median duration of hospital stay was 51 (range 14-255) days. One child died and 5 have long-term morbidity. CONCLUSIONS: The clinical features of invasive PVL-SA in this series were similar to those reported from USA and Europe. Musculoskeletal infection was the most common manifestation, frequently progressing to multiple sites and severe sepsis. Most cases had risk factors and clinical features which might have allowed earlier diagnosis, and possibly improved outcome. PMID- 19560212 TI - hCG and hyperglycosylated hCG in the establishment and evolution of hemochorial placentation. AB - The evolution of regular chorionic gonadotropin (CG) and hyperglycosylated CG are linked with the evolution of hemochorial placentation in primates. Recent research with humans shows that regular CG promotes spiral artery angiogenesis and hyperglycosylated CG controls invasion by implanting trophoblast cells. It is inferred that the evolution of regular CG and hyperglycosylated CG in early simian primates, the first species to produce these CG forms, established hemochorial placentation in this species. The circulating half-lives, and thus the circulating concentrations, of regular CG and hyperglycosylated CG increased in advanced simian primates and increased further in humans, seemingly causing greater myometrial invasion and superior angiogenesis in hemochorial placentation in advanced primates and humans. Advanced hemochorial placentation is associated with relatively high proportions of pregnancy failures in humans. This can be explained by considering human implantation inadequate in terms of invasion requirements. The demanding implantation required by the human embryo is seemingly dependent on adequate production of hyperglycosylated CG. Failures in hemochorial placentation invasion lead to anoxia and cause preeclampsia and eclampsia uniquely in humans, which can also be attributed to inadequate hyperglycosylated CG signaling. We propose here that inadequate regular CG and hyperglycosylated CG molecules are the evolutionary causes of these obstetric complications in humans. PMID- 19560213 TI - Effect of TLR3 and TLR7 activation in uterine NK cells from non-obese diabetic (NOD) mice. AB - Toll-like receptor (TLR)-TLR cross talk is thought to be important in TLR signaling. Herein, we investigated the effect of specific TLR3 and TLR7 agonists, poly (I:C) and R837, individually and in combination, on uterine immune cell function and their subsequent effects on pregnancy outcome. Allogeneic pregnancies in the non-obese diabetic (NOD) mousexC57BL/6 and wild-type BALB/cxC57BL/6 model were used. An additive increase in embryo resorption was observed after induction with both poly (I:C) and R837, and was associated with elevated numbers of both TNF-alpha- and IFN-gamma-producing CD45(+) cells in the uterus. Further examination showed that while cytokine expression was detected in both CD3(+) cells and CD49b(+) cells in BALB/c mice, NOD mouse cells behaved differently. In NOD mice, elevated cytokine expression was attributed to CD3(+) T cells, with no response detected in the CD49b(+) NK cells. The additive effect of combined agonists was partially inhibited by the Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) inhibitor SP600125 and almost completely abrogated by the extracellular signal-regulated kinase (ERK) MAPK inhibitor PD98059. These results suggest that increased TLR3 and TLR7 signals are transmitted via Th1-type T cells, rather than NK cells, in NOD mice. Furthermore, the ERK MAPK pathway may be critical in TLR3 and TLR7 signaling. PMID- 19560214 TI - Two cases of herpes zoster, occurring 4 and 6 weeks after the initiation of citalopram treatment. PMID- 19560215 TI - Acute neuroinflammation in Lewis rats - a model for acute multiple sclerosis relapses. AB - Animal models of experimental allergic encephalomyelitis (EAE) are the most commonly used animal models for studying the pathogenesis of human multiple sclerosis (MS) as well as for target validation and compound characterization in pharmacology. By using an EAE model in Lewis rats, we focus on its neuroimmunological characterization with special attention to disease-involved cytokines, chemokines, and adhesion molecules. Furthermore, we used MR imaging to investigate macrophage infiltration and ICAM-1 expression in lesional spinal cord. Overall, due to its inflammatory character, this model is suggested to be used in early drug discovery particularly directed against acute inflammatory processes. PMID- 19560216 TI - Ludwig's angina following frenuloplasty in an adolescent. AB - Ludwig's angina is a rapidly progressive cellulitis of the submandibular space and has the potential for significant upper airway obstruction. Most reported cases follow an odontogenic infection. We present the case of a 13-year-old girl who underwent a frenuloplasty to correct speech disturbances and subsequently developed a life-threatening infection of the floor of mouth. Immediate intubation, surgical decompression and antibiotic therapy successfully resolved the episode. To our knowledge, this is the first report of an iatrogenic Ludwig's angina attributable to a frenuloplasty performed for ankyloglossia. We briefly review the literature on ankyloglossia, pediatric Ludwig's angina and postoperative infections. PMID- 19560217 TI - Pattern of ear diseases in rural school children: experiences of free health camps in Nepal. AB - OBJECTIVE: This study was done to find out the prevalence of different types of ear disease in rural school children of Nepal. MATERIAL AND METHODS: This is a prospective cross-sectional study among 2000 children aged between 5 and 13 years. Children those attending free health camps in rural places of six districts of Nepal were included. These free health camps were organized either by Healthy Human Society (non-governmental organization run by doctors of Nepal) or in association with other organizations. All these children were interviewed and examined otoscopically from January 2006 to December 2008. Informed consent was taken from guardians to participate in this study. Results are expressed in numbers and percentages. RESULTS: Out of 2000 children, 64.2% were male and 35.8% female children. The most common ear diseases were wax (62.0%), followed by chronic suppurative otitis media (7.6%) and otitis media with effusion (4.7%). In CSOM, there was 83.0% tubotympanic type. Both the wax and CSOM were common in 5-7 years age group. Overall ear diseases were present in 81.6% children. CONCLUSION: Wax followed by chronic suppurative otitis media and otitis media with effusion were the most common ear diseases in rural school children of Nepal. Improvement of socioeconomic status and health care facilities, such as repetitions of free health camps will be helpful in reducing the prevalence of ear diseases. PMID- 19560218 TI - Age- and passage-dependent upregulation of fibroblast elastase-type endopeptidase activity. Role of advanced glycation endproducts, inhibition by fucose- and rhamnose-rich oligosaccharides. AB - It could be shown using the in vitro cell culture aging model, that elastase-type endopeptidase activity is progressively upregulated with successive passages (in vitro aging). Similar results were obtained previously by determining elastase type activity as a function of age in aorta extracts (human) and skin extracts (mouse). Among the possible mechanisms involved we tested the role of advanced glycation endproducts (AGEs) on this process. AGE-production was shown to increase with age, exemplified by the exponential age-dependent crosslinking of collagen, demonstrated by Fritz Verzar, already in 1963. Several AGEs significantly upregulated elastase-type activity when added to the culture medium of fibroblasts. This effect appears to be mediated by some AGE-receptors as shown previously, and could be inhibited by a 5 kDa rhamnose-rich oligosaccharide (RROP 3) as well as by a fucose-rich oligosaccharide (FROP-3). When present in the culture media, RROP-3 and FROP-3 efficiently inhibited the passage-dependent upregulation of elastase-type activity expressed by human skin fibroblasts. The use of specific inhibitors and zymography suggested that matrix metalloproteinases (MMP)-9 activation and expression are mainly involved. A detailed discussion is proposed for the interpretation of age-dependent modifications of tissues as vascular wall and skin in the light of these and related experiments, highlighting the role of several specific receptors in the mediation of the observed reactions. PMID- 19560219 TI - DNA repair in TUNEL-positive atrial cardiomyocytes of mitral and tricuspid valve diseases: potential mechanism for preserving cardiomyocytes. AB - BACKGROUND: The reported presence of DNA breaks, based on a positive reaction to the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP in situ nick end labelling (TUNEL) assay, in fibrillating human right atria of cardiac valve disease may suggest apoptotic myocyte death. However, TUNEL positivity may reflect conditions other than cell death. METHODS: This study comprised 27 adult patients (14 patients with persistent atrial fibrillation and 13 in sinus rhythm) with significant mitral and tricuspid valve diseases. Atrial tissues were obtained during surgery. RESULTS: Immunohistochemical study demonstrated that 31.1+/-12.2% of the myocytes had TUNEL-positive nuclei in the fibrillating right atria whereas 37.4+/-23.2% of the myocytes had TUNEL-positive nuclei in the right atrial myocardium in sinus rhythm (p=0.505). However, most nuclei of TUNEL positive myocytes in the right atria also expressed proliferating cell nuclear antigen (PCNA), an indicator of DNA replication and repair but never Ki-67, a replication-associated antigen (TUNEL(+)/PCNA(+) vs. TUNEL(+)/PCNA(-), 30.5+/ 10.8% vs. 1.2+/-1.5%, p=0.005, in the atrial fibrillation group and 32.8+/-18.6% vs. 4.6+/-8.1%, p=0.003, in the sinus group), suggesting that most TUNEL-positive myocytes were undergoing DNA repair. In addition, the incidence of TUNEL-positive myocytes significantly and positively correlated with the incidence of PCNA positive myocytes (r=0.5, p<0.03 in the right atria; r=0.661, p<0.04 in the left atria). CONCLUSIONS: Cell death by apoptosis occurs in a small percentage of atrial cardiomyocytes in mitral and tricuspid valve diseases and DNA repair is more important and preserves the cardiomyocytes. PMID- 19560220 TI - Optical frequency domain imaging guided crossing of a stumpless chronic total occlusion. PMID- 19560221 TI - The clinical consequences and challenges of hypertension in urban-dwelling black Africans: insights from the Heart of Soweto Study. AB - BACKGROUND: There is a paucity of data to describe advanced forms of cardiovascular disease (CVD) in urban black Africans with hypertension (HT). METHODS: Chris Hani Baragwanath Hospital services the black African community of 1.1 million people in Soweto, South Africa. We prospectively collected detailed demographic and clinical data from all de novo presentations to the hospital's Cardiology Unit in 2006. RESULTS: Overall, 761 black African patients (56% of de novo cases) presented with a diagnosis of HT with more women (63%, aged 58.5+/ 14.9 years) than men (aged 58.0+/-15.6 years). On presentation, 396 women (82%) versus 187 men (67%) had dizziness, palpitations and/or chest pain (OR 1.23, 95% 1.12-1.34: p<0.0001). HT was the primary diagnosis in 266 cases (35%). In the rest (n=495), non-ischaemic forms of heart failure were common (54% of total) while only 6.2% had coronary artery disease. Concurrent left ventricular hypertrophy, renal dysfunction and anaemia were present in 39%, 24% and 11% of cases, respectively, with a similar age-adjusted pattern of co-morbidity according to sex. However, men were more likely to present with impaired systolic function (OR 2.13, 95% CI 1.50 to 3.00; p<0.0001). CONCLUSIONS: In the absence of effective primary and secondary prevention strategies, these unique data highlight the potentially devastating impact of advanced forms of hypertensive heart disease in urban black African communities with more women than men affected. PMID- 19560222 TI - Radiation-induced lipid peroxidation activates src kinase and triggers nuclear EGFR transport. AB - PURPOSE: Elucidation of the molecular mechanism of radiation-induced activation of src kinase, which initiates EGFR internalization and nuclear transport. MATERIAL AND METHODS: Radiation-induced src activation was investigated in the bronchial carcinoma cell line A549. Proteins were Western blotted and quantified by the help of specific antibodies. Residual DNA-damage was quantified with gammaH(2)AX-foci analysis. Radiation-induced lipid peroxidation was prevented by acetyl-cysteine. RESULTS: The radiation-induced src activation and EGFR stabilization could be mimicked by addition of hydroxy-nonenal (HNE), one of the major lipid peroxidation products. Radiation-generated HNE is bound to EGFR and src and correlated with complex formation between both following radiation. Treatment with HNE activated src and stimulated radiation-associated EGFR and caveolin 1 phosphorylations resulting in increased nuclear transport of EGFR. Consequently, radiation-induced phosphorylation and activation of DNA-PK were increased. This phosphorylation was associated with improved removal of residual damage 24h after irradiation. Inhibition of radiation-induced HNE generation by acetyl-cysteine blocked radiation-induced src activation and EGFR phosphorylation. CONCLUSIONS: HNE generated in response to radiation exposure activates src kinase and is involved in regulation of radiation-stimulated DNA repair processes. PMID- 19560223 TI - The presence of pMRC01 promotes greater cell permeability and autolysis in lactococcal starter cultures. AB - Conjugative transfer of plasmid-associated properties is routinely used to generate food-grade derivatives of lactococcal starter strains with improved technological traits. However, the introduction of one or more plasmids in a single strain is likely to impose a burden on regular cell metabolism and may affect the growth characteristics of the transconjugant culture. The aim of this study was to evaluate the impact of the 60.2-kb plasmid pMRC01 (encoding for an abortive infection bacteriophage resistance system and production of the anti microbial, lacticin 3147) on starter performance. Five lactococcal strains (L. lactis HP, 255A, SK1, 712 and IL1403) and their pMRC01-containing derivatives were compared in terms of technological properties, including analysis of growth, acidification and autolysis rates. The transconjugants exhibited lower specific growth rates and higher generation times compared to the parental strains when grown at 30 degrees C in glucose-M17, but the presence of pMRC01 did not significantly affect the acidification capacity of strains in 11% reconstituted skimmed milk and synthetic media. Levels of lactate dehydrogenase were two-fold higher in supernatants of transconjugants than in those of parental strains, after 24 and 72 h of growth at 30 degrees C in glucose-M17, suggesting that the presence of pMRC01 somehow accelerates and promotes cellular autolysis. Analysis by flow cytometry following live/dead staining confirmed this result by showing larger populations of injured and dead cells in pMRC01-carrying cultures compared to the parental strains. The results of this study reveal that the plasmid pMRC01 places a burden on lactococcal host metabolism, which is associated with an increased cell permeability and autolysis, without significantly affecting the acidification capacity of the starter. While the magnitude of these effects appears to be strain dependent, the production of the bacteriocin lacticin 3147 may not be involved. PMID- 19560224 TI - NGSP and IFCC-derived NGSP HbA1c can be used interchangeably. AB - AIM: To evaluate comparability of NGSP-certified method and IFCC-calibrated method. METHODS: HbA1c was measured on two analyzers (n=50). Comparability was tested by Deming regression, bias estimation and Friedman test. RESULTS: A strong correlation and good agreement between two methods was demonstrated. CONCLUSION: NGSP and IFCC-derived NGSP HbA1c can be used interchangeably. PMID- 19560225 TI - The systemic inflammatory response syndrome in cirrhotic patients: relationship with their in-hospital outcome. AB - BACKGROUND/AIMS: Some evidence suggests that the systemic inflammatory response syndrome (SIRS) contributes to the poor outcome of cirrhotic patients. We studied 141 cirrhotic patients consecutively admitted to a tertiary referral centre assessing prevalence of SIRS and its relationship with in-hospital outcome. METHODS: Presence of SIRS was assessed on admission and during hospital stay. Main clinical outcomes were death and development of portal hypertension-related complications. RESULTS: Thirty-nine patients met SIRS criteria. SIRS was present on admission in 20 of 141 patients (14.1%), whereas it occurred during hospital stay in 19 of 121 (15.7%). SIRS was correlated with bacterial infection at admission (p=0.02), jaundice (p=0.011), high serum creatinine levels (p=0.04), high serum bilirubin levels (p=0.002), high international normalized ratio (p=0.046), high model of end-stage liver disease (MELD) score (p=0.001), and high SOFA score (p=0.003). During a follow-up of 14+/-8 days, 16 patients died (11%), 7 developed portal hypertension-related bleeding (5%), 16 hepatic encephalopathy (11%), and 5 hepatorenal syndrome type-1 (3.5%). SIRS was correlated both to death (p<0.001) and to portal hypertension-related complications (p<0.001). On multivariate analysis, SIRS and MELD were independently associated with death. CONCLUSIONS: SIRS frequently occurs in patients with advanced cirrhosis and is associated with a poor outcome. PMID- 19560226 TI - Non-alcoholic fatty liver disease in patients with chronic plaque psoriasis. AB - BACKGROUND/AIMS: Non-alcoholic fatty liver disease (NAFLD) and chronic plaque psoriasis are both associated with metabolic syndrome and increased risk of incident cardiovascular disease. We assessed the frequency and characteristics of NAFLD in patients with chronic plaque psoriasis. METHODS: One hundred and thirty consecutive patients with chronic plaque psoriasis and 260 apparently healthy controls matched for age, sex and body mass index were enrolled. NAFLD was diagnosed by abdominal ultrasound after excluding other secondary causes of chronic liver disease. RESULTS: The frequency of NAFLD was remarkably greater in psoriasis patients than in controls (47% vs. 28%; p<0.0001). Patients with psoriasis and NAFLD (n=61) were more likely to have metabolic syndrome and had higher serum C-reactive protein concentrations and greater severity of psoriasis according to the Psoriasis Area and Severity Index (PASI) score (14.2+/-12.6 vs. 9.6+/-7.4; p<0.01) than those with psoriasis alone (n=69). In a subgroup of psoriasis patients (n=43), those with NAFLD (n=21) also had significantly higher serum interleukin-6 and lower serum adiponectin levels. Notably, in multivariate regression analysis, NAFLD was associated with higher PASI score independently of age, gender, body mass index, psoriasis duration, and alcohol consumption. CONCLUSIONS: NAFLD is frequent in patients with chronic plaque psoriasis - affecting up to nearly half of these patients - and is strongly associated with psoriasis severity. Early recognition of NAFLD by radiological imaging tests in this group of patients is warranted. PMID- 19560227 TI - Health services, psychiatry and citizenship in a globalizing world: a perspective from Ireland. AB - Competing models of cultural integration present significant challenges and opportunities in the provision of public services to multi-ethnic, multi-cultural populations. In some countries, the mental health needs of refugees, voluntary migrants and ethnic minority groups are met by dedicated mental health services separate from generic services. For other countries, such developments present challenges in terms of integration objectives and public service provision in accordance with national policy. Nonetheless, given our profound, enduring attachments to culture, it appears reasonable that host societies should, at least in part, co-operate with refugees, voluntary migrants and/or ethnic minority groups in re-shaping elements of public services to optimize their appropriateness to the linguistic and historical traditions of such groups. This is particularly relevant to services that address complex states of psychological distress. In addition, it is apparent that globalization is revising received notions of citizenship and opening up political processes to groups who were previously excluded, including the mentally ill. While it is necessary to avoid excessive shifts away from the rights of the individual towards the rights of culturally defined sub-groups, it is important that public services recognize the myriad challenges and opportunities presented by evolving models of culture and citizenship in a globalizing world. PMID- 19560229 TI - Separation methods for isolation of human polymorphonuclear leukocytes affect their motile activity. AB - Five commonly used methods for the isolation of human polymorphonuclear leukocytes (PMNLs) from blood and their subsequent effect on cell motile activity were compared. Although all methods (isolation from blood clots, hemolysis of erythrocytes in hypotonic solutions, and sedimentation with the use of Percoll, Ficoll 400 or Dextran T 500 solutions) preserved cell viability, they demonstrated different effects on cell spreading and the speed of spontaneous cell movement. The highest motile activity was shown by PMNLs separated from blood clots and Percoll solutions. In the presence of formylated peptides, N-FMLP mediated movement was markedly stimulated in PMNLs separated by all five methods, but cells isolated with the use of Dextran T 500 or Ficoll 400 were relatively slower than those isolated with other methods. This suggests that the cells had preserved the sensitivity of their receptors specific for ligands stimulating chemokinesis (chemotaxis) after all five methods of separation. Immunofluorescence observations showed that PMNLs isolated in the presence of FITC-Dextran exhibited polysaccharide-coated surfaces with receptor proteins extended above that coat - an observation which explains why cell coating with polysaccharides does not disturb cell phenotyping with flow cytometry and FACS methods. PMID- 19560228 TI - Complex human chromosomal and genomic rearrangements. AB - Copy number variation (CNV) is a major source of genetic variation among humans. In addition to existing as benign polymorphisms, CNVs can also convey clinical phenotypes, including genomic disorders, sporadic diseases and complex human traits. CNV results from genomic rearrangements that can represent simple deletion or duplication of a genomic segment, or be more complex. Complex chromosomal rearrangements (CCRs) have been known for some time but their mechanisms have remained elusive. Recent technology advances and high-resolution human genome analyses have revealed that complex genomic rearrangements can account for a large fraction of non-recurrent rearrangements at a given locus. Various mechanisms, most of which are DNA-replication-based, for example fork stalling and template switching (FoSTeS) and microhomology-mediated break-induced replication (MMBIR), have been proposed for generating such complex genomic rearrangements and are probably responsible for CCR. PMID- 19560230 TI - The putative miR172 target gene InAPETALA2-like is involved in the photoperiodic flower induction of Ipomoea nil. AB - The miR172 gene is involved in the regulation of flowering time and floral organ identity in Arabidopsis thaliana through regulation of APETALA2 (AP2)-like genes' activity. AP2 plays critical roles in establishing meristem and organ identity during floral development. Additionally, the AP2-like genes including TARGET OF EAT1 (TOE1), TOE2, SMZ, SNZ are involved in the timing of flowering in Arabidopsis thaliana. In our study, a full-length cDNA encoding InAP2-like transcription factor was isolated from cotyledons of morning glory (Ipomoea nil named also Pharbitis nil), a model short day plant. The identified sequence shows significant similarity to the cDNA of TOE1 from Arabidopsis thaliana and contains nucleotides complementary to miR172. Semi-quantitative RT-PCR analysis and in situ hybridization showed that the accumulation of InAP2-like transcripts was high, especially in cotyledons of 5-d-old seedlings. During the 16h-long inductive night, an increase in the expression of InAP2-like and a decrease in the accumulation of miR172 were observed. Auxin and ethylene treatment, as well as a "night-break", which completely eliminated flowering induction of Ipomoea nil, caused a decrease in the InAP2-like mRNAs levels in cotyledons of Ipomoea nil. These results suggest the potential involvement of miR172 and InAP2-like in the mechanism of flowering induction in Ipomoea nil. PMID- 19560231 TI - The Spanish national health care-associated infection surveillance network (INCLIMECC): data summary January 1997 through December 2006 adapted to the new National Healthcare Safety Network Procedure-associated module codes. AB - BACKGROUND: In 1997, a national standardized surveillance system (designated INCLIMECC [Indicadores Clinicos de Mejora Continua de la Calidad]) was established in Spain for health care-associated infection (HAI) in surgery patients, based on the National Nosocomial Infection Surveillance (NNIS) system. In 2005, in its procedure-associated module, the National Healthcare Safety Network (NHSN) inherited the NNIS program for surveillance of HAI in surgery patients and reorganized all surgical procedures. METHODS: INCLIMECC actively monitors all patients referred to the surgical ward of each participating hospital. We present a summary of the data collected from January 1997 to December 2006 adapted to the new NHSN procedures. RESULTS: Surgical site infection (SSI) rates are provided by operative procedure and NNIS risk index category. Further quality indicators reported are surgical complications, length of stay, antimicrobial prophylaxis, mortality, readmission because of infection or other complication, and revision surgery. CONCLUSION: Because the ICD-9-CM surgery procedure code is included in each patient's record, we were able to reorganize our database avoiding the loss of extensive information, as has occurred with other systems. PMID- 19560234 TI - Nursing professional values: validation of a scale in a Spanish context. AB - PURPOSE: To validate culturally the "Nursing Professional Values Scale: NPVS-R" for use in Spain. METHODS: We used the forward and backward translation method with complementary qualitative methodology. We first formed discussion groups to detect cultural differences, then set up a multidisciplinary group to ascertain the equivalence or discrepancies between the English and Spanish versions and to decide on the final pre-test version. This was used in the pilot test given to 10 second-year nursing students of the School of Nursing of Vitoria, Spain. RESULTS: The majority of the questions were classified as equivalents (23 of 26). Some questions were modified by the discussion group. Following the backward translation and evaluation by the multidisciplinary group, two questions needed small changes and four needed significant changes. The differences were resolved by decentralization. CONCLUSIONS: The resulting Spanish version of the NPVS-R, is culturally and semantically equivalent to the original English version and is appropriate for measuring the perception of nursing professional values in nursing professionals and students in Spain. To verify reliability and validity of the scale, the final version after the pilot test has been passed to 960 nursing diploma students all over Spain. PMID- 19560233 TI - HFE polymorphisms affect cellular glutamate regulation. AB - HFE gene variants are relatively common genetic variants in Caucasians. The H63D HFE genetic variant has been repeatedly associated with a number of neurodegenerative diseases. We developed neuroblastoma cell lines expressing different HFE polymorphisms to explore the mechanisms behind these associations. Here we tested the hypothesis that cells with the H63D variant have a phenotype that promotes glutamate toxicity. In support of this hypothesis, expression of H63D HFE is associated with increased calcium-induced glutamate secretion and decreased cellular glutamate uptake. The polymorphism-associated changes in glutamate secretion were mimicked by altering cellular iron. Additionally, intracellular calcium is altered in a genotype-specific manner which could further impact glutamate secretion. HFE-dependent effects on glutamate uptake were confirmed in astrocytoma cell lines with endogenous expression of HFE. The ability of minocycline and the antioxidant Trolox to increase glutamate uptake differed by HFE genotype and implicate oxidative stress in glutamate regulation. This study demonstrates HFE cellular effects that extend beyond iron regulation, and suggests that H63D HFE may promote glutamate toxicity. PMID- 19560232 TI - Incidental Lewy body disease: do some cases represent a preclinical stage of dementia with Lewy bodies? AB - Lewy pathology occurs in 8-17% of neurologically normal people age >60, termed incidental Lewy body disease (iLBD). It is often assumed to represent preclinical Parkinson disease (PD). However, some iLBD cases have diffuse pathology inconsistent with preclinical PD. We analyzed iLBD cases (alpha-synuclein immunohistochemistry) using the Braak PD staging scheme and determined if some had a neuropathological pattern suggestive of preclinical dementia with Lewy bodies (DLB). Of the 235 brains examined, 34 had iLBD (14.5%) and all but one could be assigned a Braak PD stage. The distribution of alpha-synuclein pathology in the 33 cases fell into three patterns: (1) diffuse cortical and subcortical alpha-synuclein pathology; (2) no cortical alpha-synuclein pathology, but a caudal-to-rostral ascending pattern, primarily involving brainstem; and (3) intermediate between these two categories. Also, 6/33 cases failed to follow the pattern of contiguous spread proposed by Braak. These findings suggest dichotomy in the distribution of iLBD: some cases fit the Braak ascending scheme, conceptually consistent with preclinical PD, whereas others displayed prominent cortical involvement that might represent preclinical DLB. PMID- 19560235 TI - Nursing education in Turkey: from past to present. AB - Nursing education in Turkey first began in 1912 with the introduction of a 6 month course to train voluntary medical attendants, with Dr. Besim Omer Pasha's advice to the Red Crescent Association regarding the inadequacy of healthcare services, as a crucial need for nursing services resulted due to significant losses given during Tripoli (1911) and the Balkan (1912) wars. Carrying out their duties in battlefields and hospitals with great devotion, the first nurses graduated from the course played a significant role in promoting the nursing profession and its importance. Nursing education which used to continue at the secondary and high-school levels increased to bachelor's level in 1955. Master's program in nursing was opened in 1968, and Ph.D. programs was opened in 1972. Professional members of the practice, well-equipped in accordance with the requirements of the age, who conduct their studies at the national and international levels, are trained as the consequence of the recent developments in nursing education. The number of nurses at the universities who offered higher levels of academic degree, and especially the number of nurses who gained 'science expert' title at the inpatient medical establishments has increased. This situation and globalization, which ensures an easier access to nursing literature through internet, enable a more systematic and of a better quality healthcare. This article explains the nursing education in Turkey from past to present. The developments in nursing education which have taken place in Turkey are expressed in a chronological order, starting from the Ottoman Empire, until the present. Compared with other countries, nursing education is given on different levels in Turkey. Recently, however, the obstacles regarding the differences especially at the bachelor's degree level were overcome, appropriate changes were made, and education melioration efforts gained speed. PMID- 19560236 TI - Not choosing nursing: work experience and career choice of high academic achieving school leavers. AB - Work experience has been a feature of the secondary school curriculum in the United Kingdom for a number of years. Usually requested by the pupil, it aims to provide opportunities for school pupils to enhance their knowledge and understanding of an occupation. The main benefits are claimed to be that it can help pupils develop an insight into the skills and attitudes required for an occupation and an awareness of career opportunities. However the quality and choice of placements are considered to be of great importance in this process and in influencing career choice [Department for Education and Skills (DfES), 2002a. Work Experience: A Guide for Employers. Department for Education and Skills, London]. As university departments of nursing experience a decline in the number of school pupils entering student nurse education programmes, and with the competition for school leavers becoming even greater, it is important to consider whether school pupils have access to appropriate work placements in nursing and what influence their experience has on pursuing nursing as a career choice. This paper is based on interview data from 20 high academic achieving fifth and sixth year school pupils in Scotland, paradigmatic cases from a larger survey sample (n=1062), who had considered nursing as a possible career choice within their career preference cluster, but then later disregarded nursing and decided to pursue medicine or another health care profession. This was partly reported by Neilson and Lauder [Neilson, G.R., Lauder, W., 2008. What do high academic achieving school pupils really think about a career in nursing: analysis of the narrative from paradigmatic case interviews. Nurse Education Today 28(6), 680 690] which examined what high academic achieving school pupils really thought about a career in nursing. However, the data was particularly striking in revealing the poor quality of nursing work experience for the pupils, and also their proposal that there was a need for work experience which was more representative of the reality of nursing. Participants reported that proper work experience in nursing could make it more attractive as a career choice but that there were difficulties and barriers in obtaining an appropriate work experience in nursing. These included unhelpful attitudes of teachers towards work experience in nursing in general and the placements themselves which were typically in a nursing home or a care home. They felt that departments of nursing within universities should have an input into organising more realistic work placements and that their involvement could foster greater interest amongst pupils in nursing as a career. PMID- 19560237 TI - Involving the stakeholders in the curriculum process: a recipe for success? AB - The Department of Nursing and Health Studies at the Manukau Institute of Technology (MIT) in Auckland, New Zealand, decided to involve stakeholders from the health care sector in developing a new curriculum. After implementing the new curriculum, the process was evaluated using a content analysis as qualitative research design. Seven individual interviews and one Focus group interview were conducted with the stakeholders to determine their experiences during the process. Ethical permission was sought from the MIT ethical committee. The analyses of the collected data enabled the researchers to identify six main categories. The categories were: "Existing Programme", "The need to change", "The curriculum development process", "The stakeholders", "Personnel", and "Ethnic minorities". From the collected data, it was clear that a new curriculum was necessary to enable the graduates to meet the health care needs of the New Zealand population, especially after the primary health care policy was introduced in New Zealand. It was also clear that the curriculum development process could be a painful process for all concerned, but a strong leadership could cement a feeling of "collegiality" between stakeholders and teaching staff. The importance of considering the rights of ethnic minorities is clearly stated in the Treaty of Waitangi, safeguarding the rights of the Maori People, and therefore applied rigorously in the development process. In this project, the collaborative process was very successful, and the stakeholders actually expressed feelings of "Ownership" of the curriculum. PMID- 19560238 TI - Cutoff values for central obesity in Chinese based on mesenteric fat thickness. AB - AIMS: Sonographic measurement of mesenteric fat thickness (MFT) is a novel, accurate and simple tool to evaluate regional distribution of obesity. We used MFT to determine the optimal waist circumference (WC) values and associated risk factors for cardiovascular disease (CVD). METHODS: 282 healthy Chinese (age 41.8+/-7.4 years, BMI 23.8+/-3.3 kg/m(2)) was assessed. High MFT was defined as mean+1 SD of the cohort. We compared the CVD risks including fatty liver amongst subjects with normal waist, central pre-obesity and central obesity. RESULTS: WC of 84.6 cm in men and 75.7 cm in women were the optimal cutoff values to predict high MFT with ROC analysis. Using WC cutoff values > or =85-90 cm and > or =90 cm to define central pre-obesity and obesity in men (> or =75-80 cm and > or =80 cm in women), both central obesity and pre-obesity had higher MFT and CVD risk than those with normal waist. The frequencies of fatty liver in these 3 categories were 15.9%, 56.7% and 96.7% in men and 6.9%. 17.9% and 63.2% in women (p<0.001 for trend). CONCLUSION: In addition to central obesity, "central pre-obesity" identifies subjects who harbor high CVD risks, fatty liver and excess visceral fat. PMID- 19560239 TI - Aspergillosis, maxillary foreign body, and mercury amalgam. PMID- 19560240 TI - Re: The vertical component of the ground reaction force does not reflect horizontal braking or acceleration per se. PMID- 19560241 TI - Tibiofemoral contact mechanics with a femoral resurfacing prosthesis and a non functional meniscus. AB - BACKGROUND: Increased contact stress with a femoral resurfacing prosthesis implanted in the medial femoral condyle and a non-functional meniscus is of concern for potential deleterious effects on tibiofemoral contact mechanics. METHODS: Peak contact pressures were determined in seven fresh frozen human cadaveric specimens using a pressure sensitive sensor placed in the medial compartment above the menisci. A knee simulator was used to test each knee in static stance positions (5 degrees/15 degrees/30 degrees/45 degrees) and through 10 dynamic knee flexion cycles (5-45 degrees) with single body weight ground reaction force which was adjusted to the living body weight of the cadaver donor. All specimens were tested in three different conditions: untreated knee (A); flush implantation of a 20mm resurfacing prosthesis (HemiCAP) in the weight bearing area of the medial femoral condyle (B); complete radial tear at the posterior horn of the medial meniscus with the femoral resurfacing device in place (C). FINDINGS: On average, flush device implantation resulted in no statistically significant differences when compared to the untreated normal knee. The meniscal tear resulted in a significant increase of the mean maximum peak contact pressures by 63%, 57%, and 57% (all P< or =0.05) at 15 degrees , 30 degrees and 45 degrees static stance positions and 78% (P< or =0.05) through the dynamic knee flexion cycle. No significant different maximum peak contact pressures were observed at 5 degrees stance position. INTERPRETATION: Although the condition of a meniscal tear without the resurfacing device could not be compared, possible effects of reduced meniscal tissue and biomechanical integrity of the meniscus must be considered in an in vivo application. PMID- 19560242 TI - From "first" to "last" steps in life--pressure patterns of three generations. AB - BACKGROUND: The human foot has to bear loads during all kinds of bipedal locomotion throughout the whole life. Rapid developmental changes of foot morphology and foot function occur during the first years of walking. Furthermore, disease dependent modifications can also have an influence on plantar loading. Therefore, it is reasonable to assume that foot function will undergo changes in life. However, the main differences between the pressure patterns in young and elderly have not been well described. The aim of the study was to evaluate age-dependent pressure patterns in different age-related stages. METHODS: Hundred and four healthy humans of four different age groups were retrospectively analysed by means of plantar pressure measurements (toddlers: mean age 1.0 (SD 0.2) year; 7-year olds: 7.0 (SD 0.4) years; adults: 31.9 (SD 2.1) years; seniors: 68.7 (SD 3.2) years). The emed pressure platform was used to evaluate peak pressure, maximum force, contact time, contact area and arch index. FINDINGS: Significant differences were found for each parameter between almost every age group. The highest peak pressure values were observed for the seniors' (P<0.001). Peak pressures are low in toddlers (145 kPa), high in 7-year olds and adults (400-600 kPa) and even higher in elderly (> or =800 kPa). INTERPRETATION: Elderly adults can still be functionally mobile even if pressures are high. The results for the investigated age groups can be used as normative foot loading data to compare to pathological foot function. PMID- 19560243 TI - The algal toxicity of silver engineered nanoparticles and detoxification by exopolymeric substances. AB - In this study, we report that silver ions (Ag(+)) from the oxidative dissolution of silver engineered nanoparticles (Ag-ENs) determined the EN toxicity to the marine diatom Thalassiosira weissflogii. Most of the Ag-ENs formed non-toxic aggregates (>0.22 microm) in seawater. When the free Ag(+) concentration ([Ag(+)](F)) was greatly reduced by diafiltration or thiol complexation, no toxicity was observed, even though the Ag-ENs were better dispersed in the presence of thiols with up to 1.08 x 10(-5) M Ag-ENs found in the <0.22 microm fraction, which are orders of magnitude higher than predicted for the natural aquatic environment. The secretion of polysaccharide-rich algal exopolymeric substances (EPS) significantly increased at increasing [Ag(+)](F). Both dissolved and particulate polysaccharide concentrations were higher for nutrient-limited cells, coinciding with their higher Ag(+) tolerance, suggesting that EPS may be involved in Ag(+) detoxification. PMID- 19560244 TI - HIV disclosure and unsafe sex among HIV-infected women in Cameroon: results from the ANRS-EVAL study. AB - Encouraging seropositive people to voluntarily disclose their serostatus has been promoted as a key component of HIV prevention. Among other reasons, HIV disclosure to one's main partner is believed to be an incentive for serodiscordant couples to practice safe sex. The present article investigated this issue by conducting a cross-sectional survey of a large sample of HIV infected women attending HIV care centers in Cameroon (N=1014). Overall, 86.3% of these women had disclosed their serostatus to their main partner. With respect to sexual activity with their main partner during the previous three months, 35.0% had practiced abstinence and 47.4% only safe sex, whereas 17.6% had engaged in unsafe sex at least once. HIV disclosure to one's main partner was related to safe sexual practices in multivariate analysis. Some of the factors associated with disclosure and safe sex were illustrative of the positive roles of improved access to care and women's empowerment. On the contrary, beliefs overestimating the effectiveness of antiretroviral therapies were quite widespread among respondents and predictive of both concealment of HIV status and unsafe sex. PMID- 19560245 TI - Threats to safe motherhood in Honduran Miskito communities: local perceptions of factors that contribute to maternal mortality. AB - Despite global initiatives to lower rates of maternal death, barriers to safe motherhood persist, particularly in socially and economically marginalized communities. This article describes the risks that women in Honduran Miskito villages encounter during pregnancy and childbirth. Ethnographic data are used to examine emic understandings of the underlying causes of maternal death. Participant observation, four community discussions, individual interviews with 218 women and five midwives, and a maternal mortality survey were conducted during November 2004 through November 2005. Case studies are drawn from the 55 death histories collected during the survey to illustrate the factors that contribute to maternal mortality. Community members identified poverty, gender inequality, witchcraft, and sorcery as major threats to safe motherhood. All of these factors influence women's health-related behaviors; and therefore, each issue deserves attention from public health officials. Designing appropriate interventions to improve maternal health depends on understanding the forces that increase women's vulnerability during pregnancy and childbirth. Local perspectives of risk, even when they diverge from biomedical understandings, point to specific needs, issues to address, and avenues for effective intervention. PMID- 19560246 TI - Racialized identity and health in Canada: results from a nationally representative survey. AB - This article uses survey data to investigate health effects of racialization in Canada. The operative sample was comprised of 91,123 Canadians aged 25 and older who completed the 2003 Canadian Community Health Survey. A "racial and cultural background" survey question contributed a variable that differentiated respondents who identified with Aboriginal, Black, Chinese, Filipino, Latin American, South Asian, White, or jointly Aboriginal and White racial/cultural backgrounds. Indicators of diabetes, hypertension and self-rated health were used to assess health. The healthy immigrant effect suppressed some disparity in risk for diabetes by racial/cultural identification. In logistic regression models also containing gender, age, and immigrant status, no racial/cultural identifications corresponded with significantly better health outcomes than those reported by survey respondents identifying as White. Subsequent models indicated that residential locale did little to explain the associations between racial/cultural background and health and that socioeconomic status was only implicated in relatively poor health outcomes for respondents identifying as Aboriginal or Aboriginal/White. Sizable and statistically significant relative risks for poor health for respondents identifying as Aboriginal, Aboriginal/White, Black, Chinese, or South Asian remained unexplained by the models, suggesting that other explanations for health disparities by racialized identity in Canada - perhaps pertaining to experiences with institutional racism and/or the wear and tear of experiences of racism and discrimination in everyday life - also deserve empirical investigation in this context. PMID- 19560247 TI - "AIDS is rape!" gender and sexuality in children's responses to HIV and AIDS. AB - This paper examines young African school children's understanding of HIV and AIDS. Based on focus group interviews with children aged 7-8 in KwaZulu-Natal province, South Africa, it explores the ways in which gender and sexuality feature in their responses to the disease. Data were collected between 2003 and 2004 through 26 focus groups involving 55 boys and 64 girls. The paper argues that younger children are active agents in giving meaning to the disease. Their agency is negotiated within complex social processes involving sexual violence, highly unequal gender/age inequalities, but also sexual expression. Those expressions are subsumed however under a regime of violence and fear catapulting men, albeit with contestation, as chief vectors in the spread of the disease and a source of girls' anxieties. Children's responses to the disease are the effects of material, symbolic and discursive forces effectively constraining the opportunities available to them and creating patterns of vulnerability especially for young girls. Interventions aimed at scaling up efforts to address young children responses to the disease must be situated in parallel efforts to end poverty, sexual violence and pervasive gender inequalities in order to foster more comprehensively the exercise of young children's agency. PMID- 19560249 TI - Virtues of simple hydro-economic optimization: Baja California, Mexico. AB - This paper uses simple hydro-economic optimization to investigate a wide range of regional water system management options for northern Baja California, Mexico. Hydro-economic optimization models, even with parsimonious model formulations, enable investigation of promising water management portfolios for supplying water to agricultural, environmental and urban users. CALVIN, a generalized hydro economic model, is used in a case study of Baja California. This drought-prone region faces significant challenges to supply water to agriculture and its fast growing border cities. Water management portfolios include water markets, wastewater reuse, seawater desalination and infrastructure expansions. Water markets provide the flexibility to meet future urban demands; however conveyance capacity limits their use. Wastewater reuse and conveyance expansions are economically promising. At current costs desalination is currently uneconomical for Baja California compared to other alternatives. Even simple hydro-economic models suggest ways to increase efficiency of water management in water scarce areas, and provide an economic basis for evaluating long-term water management solutions. PMID- 19560250 TI - Alternative approaches to the construction of a composite indicator of agricultural sustainability: An application to irrigated agriculture in the Duero basin in Spain. AB - This paper describes a comparative analysis of alternative methods of constructing composite indicators to measure the sustainability of the agricultural sector. The three methods employed were Principal Component Analysis, the Analytic Hierarchy Process and a Multi-Criteria technique. The comparison focused on the irrigated agriculture of the Duero basin in Spain as a case study, using a dataset of indicators previously calculated for various farm types and policy scenarios. The results enabled us to establish a hierarchy of preferred policy scenarios on the basis of the level of sustainability achieved, and show that the most recent CAP reform is the most sustainable agricultural policy scenario. By analyzing the heterogeneity of different farms types in each scenario, we can also determine the main features of the most sustainable farms in each case. The analysis demonstrates that full-time farmers with small to medium-sized farms and sowing profitable crops are the most sustainable farm types in all the policy scenarios. All of this information is useful for the support of agricultural policy design and its implementation, as we attempt to improve the sustainability of this sector. PMID- 19560248 TI - Electrophysiological evidence for size invariance in masked picture repetition priming. AB - This experiment examined invariance in object representations through measuring event-related potentials (ERPs) to pictures in a masked repetition priming paradigm. Pairs of pictures were presented where the prime was either the same size or half the size of the target object and the target was either presented in a normal orientation or was a normal sized mirror reflection of the prime object. Previous masked repetition priming studies have found a cascade of priming effect sensitive to perceptual (N190/P190) and semantic (N400) properties of the stimulus. This experiment found that both early (N190/P190 effects) and later effects (N400) were invariant to size, whereas only the N190/P190 effect was invariant to mirror reflection. The combination of a small prime and a mirror reflected target led to no significant priming effects. Taken together, the results of this set of experiments suggests that object recognition, more specifically, activating an object representation, occurs in a hierarchical fashion where overlapping perceptual information between the prime and target is necessary, although not always sufficient, to activate a higher level semantic representation. PMID- 19560251 TI - Minimally invasive ultrasound method for intra-articular diagnostics of cartilage degeneration. AB - Quantitative ultrasound imaging (QUI) can be used to evaluate the integrity of articular cartilage and for diagnosing the early signs of osteoarthritis (OA). In this study, we applied a minimally invasive ultrasound imaging technique and investigated its ability to detect superficial degeneration of bovine knee articular cartilage. Intact (n=13), collagenase-digested (n=6) and mechanically degraded (n=7) osteochondral samples (dia.=25 mm) and custom-made phantoms with different degrees of surface roughness (n=8) were imaged using a high-frequency (40 MHz) QUI system. For each sample and phantom, the ultrasound reflection coefficient (R), integrated reflection coefficient (IRC) and ultrasound roughness index (URI) were determined. Furthermore, to evaluate the clinical applicability of intra-articular ultrasound (IAUS) in diagnostics, one intact bovine knee joint was investigated ex vivo using a simulated arthroscopic approach. Differences in the surface characteristics of the phantoms were detected by monitoring changes in the reflection and surface roughness parameters. Both mechanically- and enzymatically-induced degradation were sensitively diagnosed by decreased (p<0.05) reflection (R and IRC) at the cartilage surface. Furthermore, mechanical degradation was detected in the increased (p<0.05) surface roughness (URI). The intra-articular investigation of a bovine knee joint suggested that the IAUS technique may enable minimally invasive, straightforward diagnostics of the degenerative status of the articular surfaces. We conclude that quantitative IAUS imaging can be used for detecting collagen disruption and increased roughness of the articular surface. This quantitative in vivo ultrasound technique could have great clinical value in the diagnostics of joint diseases. PMID- 19560252 TI - The automated assessment of ultrasound scanner lateral and slice thickness resolution: use of the step response. AB - The imaging performance assessment of ultrasound scanners based on traditional phantoms is limited by repeatability, subjectivity and systematic errors giving low confidence in results. A new approach to the automated measurement of scanner resolution is described. The method utilises a step change in backscatter to derive resolution from the imaging system line spread function and has been used to calculate resolution in two dimensions as a continuous function of depth. Resolution data was used to calculate resolution integrals for both lateral and slice thickness independently. For resolution integral repeatability, analysis of variance showed no significant difference between operators (p=0.05) with intra and inter-operator repeatability (+/-1 standard deviation) of 1.5% and 1.5% for lateral resolution, respectively, and 2.6% and 3.3% for slice thickness, respectively. Low contrast penetration was also calculated automatically and the worst case operator repeatability was 1.3%. The acoustic properties of the phantom were validated. The possibility of extending the technique to axial resolution is discussed. PMID- 19560253 TI - Comparison of degree of stenosis and plaque volume for the assessment of carotid atherosclerosis using 2-D ultrasound. AB - The degree of carotid stenosis (%ST) remains the most frequently used parameter for identifying patients with high risk of stroke but the relationship between %ST and the occurrence of stroke remains controversial. The objectives of this study were to check (1) the relationship between the %ST and the plaque volume index (PVI) as measured by echography and Doppler, (2) the relationship between the intima media thickness (IMT), a vessel wall remodeling index and the PVI an atheromatous growth index. For each of the 128 patients, (165 carotid stenosis), we measured the % ST (section or diameter), the max stenosis velocity (V(max)), the PVI and the common carotid IMT. The %ST (section) ranged from 10% to 93% (mean 66+/-18), V(max) from 0.3m/s to 3m/s (mean 1.2+/-0.8), PVI from 0.61cm(3) to 1.17cm(3) (mean 0.41+/-0.21) and the IMT from 0.08cm up to 0.31cm (mean 0.12+/ 0.03). There was no significant correlation between either PVI and %ST (section or diameter), PVI and minimal stenosis section area (S1) or between PVI and V(max). There was no significant correlation between IMT and both %ST area and PVI. PVI was significantly correlated with the whole artery section area (S2) and the plaque length (L). The %ST (section or diameter) was significantly correlated with S1 but not with S2. The absence of correlation between the PVI and the %ST confirm that these two parameters describe two different processes of the atheromatous development. PMID- 19560254 TI - Therapeutic potential of Mdm2 inhibition in malignant germ cell tumours. AB - BACKGROUND: Inadequate response to cisplatin-based chemotherapy is associated with poor prognosis in patients with advanced malignant testicular germ cell tumours (TGCTs), especially of the nonseminomatous type. Novel chemotherapeutic agents have failed so far to significantly improve the outcome of such patients. The majority of these tumours express low levels of p53, and TP53 mutations are rarely observed. Murine double minute 2 (Mdm2) inhibitors enhance apoptosis in tumours harbouring wild-type p53. OBJECTIVE: We sought to investigate the potential therapeutic value of Mdm2 in TGCT-derived cell lines with the histology of nonseminoma. DESIGN, SETTING, AND PARTICIPANTS: The Mdm2 inhibitor nutlin-3 was evaluated alone and in combination with cisplatin in a panel of germ cell tumour (GCT)-derived cell lines (embryonal carcinomas, being the nonseminomatous stem-cell component) with wild-type (NT2 and 2102EP cells) and mutant (NCCIT cells) p53 status. MEASUREMENTS: Biological consequences of Mdm2 inhibition were determined by analysis of the p53 pathway, cell proliferation, and apoptosis. RESULTS AND LIMITATIONS: Nutlin-3 exhibited significant activity (IC50 2.8 MUM) in NT2 and 2102EP (wild-type p53) but not in p53-mutant NCCIT cells (<10% inhibition at 10 MUM). At concentrations beyond 500 nM, additive effects were seen for the combination of nutlin-3 and cisplatin in NT2 and 2102EP cells but not in NCCIT cells. This correlated with the induction of p53 and its target p21, suggesting an on-target effect of nutlin-3. Moreover, nutlin-3 (5 MUM) and cisplatin (0.5 MUM) additively induced caspase cleavage and apoptosis in NT2 cells and 2102-EP cells but not in p53-mutant NCCIT cells. CONCLUSIONS: These results provide strong evidence for further development of pharmacologic Mdm2 inhibition for the treatment of patients suffering from high-risk nonseminomatous TGCT with wild-type p53 status. PMID- 19560255 TI - A comparison of postoperative complications in open versus robotic cystectomy. AB - BACKGROUND: Robotic cystectomy is an emerging alternative for treatment of invasive bladder cancer (BCa). However, reduction in postoperative morbidity relative to the open approach has not been demonstrated. OBJECTIVE: To compare complication rates in patients undergoing robotic versus open radical cystectomy (RC). DESIGN, SETTING, AND PARTICIPANTS: A prospective cohort study of 187 consecutive patients undergoing RC at our institution-104 open RC, 83 robotic RC. INTERVENTION: Open or robotic RC with urinary diversion. MEASUREMENTS: Demographic, perioperative, and complication data were recorded prospectively. Thirty-day and 90-d complication rates were assessed using the modified Clavien complication scale. Data were evaluated using chi(2) and multivariate logistic regression analyses. RESULTS AND LIMITATIONS: At 30 d, the open group demonstrated a higher overall complication rate (59% vs 41%; p=0.04) as well as more major complications (30% vs 10%; p=0.007). At 90 d, the overall complication rate was greater in the open group, but this was not statistically significant (62% vs 48%; p=0.07). However, there was a significantly higher major complication rate in the open cohort (31% vs 17%; p=0.03). When subjected to logistic regression analysis, robotic cystectomy was an independent predictor of fewer overall and major complications at 30 and 90 d. High American Society of Anesthesiologists (ASA) score (3-4) and longer surgical time were independent predictors of major complications. Though this is one of the largest published RC series, the sample size is relatively small. Moreover, despite the two patient cohorts being similarly matched, the study was not performed in a randomized fashion. CONCLUSIONS: Patients undergoing robotic cystectomy experienced fewer postoperative complications than those undergoing open cystectomy. Robotic cystectomy is an independent predictor of fewer overall and major complications. Until long-term oncologic results are available, robotic cystectomy should still be considered investigational. PMID- 19560256 TI - Editorial comment on: a comparison of postoperative complications in open versus robotic cystectomy. PMID- 19560257 TI - Single postoperative instillation of gemcitabine in patients with non-muscle invasive transitional cell carcinoma of the bladder: a randomised, double-blind, placebo-controlled phase III multicentre study. AB - BACKGROUND: Recurrence prophylaxis with intravesical gemcitabine (GEM) was effective and safe in patients with non-muscle-invasive bladder cancer (NMIBC); efficacy as single-shot instillation remains to be proved. OBJECTIVE: To compare the efficacy of a single GEM instillation versus placebo (PBO) immediately after transurethral resection (TUR) of tumour in patients with histologically confirmed NMIBC (pTa/pT1,G1-3). DESIGN, SETTING, AND PARTICIPANTS: This was a double-blind, randomised, PBO-controlled study in patients with clinical evidence of primary or recurrent NMIBC (Ta/T1,G1-3). Of 355 patients randomised at 24 urologic centres, 328 underwent TUR and received instillation (92.4%; GEM/PBO: 166/162). In case of nonmalignancy, carcinoma in situ (CIS), > or = pT2 disease, or intraoperative complications, patients were discontinued. INTERVENTION: We used a single, postoperative 30-40-min instillation of GEM (2000 mg/100 ml of saline) or PBO (100 ml of saline) followed by continuous bladder irrigation for > or = 20 h. A second TUR (no instillation) and adjuvant bacillus Calmette-Guerin (BCG) instillations were allowed. MEASUREMENTS: Primary outcome was recurrence-free survival (RFS). Secondary outcomes included type of recurrence and adverse events. To detect a difference in RFS, 191 recurrences were required (80% power, log-rank-test, alpha = 0.050). RESULTS AND LIMITATIONS: Two hundred forty-eight patients (69.9%, GEM, PBO: 124, 124) had histologically confirmed pTa/pT1 G1-3 Gx tumour and were eligible for efficacy (GEM: 76.6% male; median age: 65 yr; PBO: 83.1% male; median age: 67 yr). Treatment groups were balanced (pTa: 75.0%, 71.0%; G1-G2: 85.5%, 87.9%; recurrent tumour: 24.2%, 21.0%; BCG: 10.5%, 16.9%). After a median follow-up of 24 mo, there were only 94 recurrences and 11 deaths. The study was terminated early based on predefined decision criteria. RFS was high in both groups (12-mo RFS [95% confidence interval (CI)]: GEM: 77.7% [68.8 84.3]; PBO: 75.3% [66.3-82.3]). There was no significant group difference (hazard ratio [HR]: 0.946 [0.64-1.39], log-rank test, p=0.777). CONCLUSIONS: In this study of NMIBC, the immediate single instillation of GEM 2000 mg/100 ml of saline after TUR was not superior to PBO in terms of RFS. Rigid continuous irrigation and improved TUR/cystoscopy techniques may have contributed to the high RFS in both groups. PMID- 19560258 TI - Long-term survival in patients undergoing radical nephrectomy and inferior vena cava thrombectomy: single-center experience. AB - BACKGROUND: Renal cell carcinoma (RCC) with a tumor thrombus extension into the inferior vena cava (IVC) demands aggressive surgical management. OBJECTIVE: To evaluate the long-term survival in patients undergoing radical nephrectomy and IVC thrombectomy. DESIGN, SETTING, AND PARTICIPANTS: We performed a retrospective analysis of 87 patients undergoing surgery between 1997 and 2008. The patients were grouped according to the extent of tumor thrombus, with level I involving the IVC at the level of the renal vein, level II being infrahepatic IVC, level III being intrahepatic IVC, and level IV being suprahepatic IVC or right atrium. Relevant clinical and pathologic data were analyzed. MEASUREMENTS: Disease-free survival (DFS) and disease-specific survival (DSS) were studied. RESULTS AND LIMITATIONS: The median follow-up was 22 mo, and 19, 14, 40, and 14 patients had level I, II, III, and IV IVC thrombus, respectively. Among patients with M0 disease, 22 developed metastases. The 5-yr DFS was 64% for all levels and 74%, 69.5%, 59.5%, and 58% for levels I, II, III, and IV, respectively. Of the level I group, 16% of patients died of disease compared to 57% of the level IV group. The 5-yr DSS for all levels was 46% and 71%, 48%, 40%, and 35% for levels I, II, III, and IV, respectively. Patients with level IV thrombus had a significantly lower 5 yr DSS compared to level I (p=0.03). However, when analyzed in two groups supradiaphragmatic and infradiaphragmatic-there was no significant difference in DSS (P=0.14). On univariate analysis, metastasis at presentation, non-clear-cell histology, lymph node metastases, and higher nuclear grade were statistically significant prognostic factors influencing DSS. Only higher nuclear grade (p=0.03), metastasis at presentation (p<0.01), and non-clear-cell histology (p=0.03) were independent prognostic factors on multivariate analysis. CONCLUSIONS: Radical nephrectomy and IVC thrombectomy offer reasonable long-term survival. The level of tumor thrombus is not an independent prognostic factor. Distant metastasis at presentation, higher nuclear grade, and non-clear-clear cell histology are significant prognostic factors influencing DSS. PMID- 19560259 TI - Laparoscopic versus open nephroureterectomy: perioperative and oncologic outcomes from a randomised prospective study. AB - BACKGROUND: Laparoscopic nephroureterectomy (LNU) is increasingly being used instead of open nephroureterectomy (ONU) for the treatment of urothelial carcinoma (UC) of the upper urinary tract (UUT), but the evidence of equal oncologic effectiveness is still lacking. OBJECTIVE: To present perioperative and oncologic results from a prospective randomised study comparing ONU and LNU. DESIGN, SETTING, AND PARTICIPANTS: Eighty patients with nonmetastatic UUT UC and without previous history of UC were enrolled. Of those, 40 patients (group A) randomly received ONU and 40 patients (group B) randomly received LNU. INTERVENTIONS: ONU was performed through a flank incision with a lower quadrant incision to allow excision of a bladder cuff. Transperitoneal LNU was performed with a four-trocar technique, and bladder cuff was detached with a 10-mm LigaSure device. MEASUREMENTS: Perioperative data were compared with the student t test. Bladder tumour-free survival (BTFS), metastasis-free survival (MFS), and cancer specific survival (CSS) curves for both groups were compared with the log-rank test before and after stratifying patients for pT category and tumour grade. RESULTS AND LIMITATIONS: Operative times were comparable, while mean blood loss and mean time to discharge were significantly lower in group B (both p values <0.001). At a median follow-up of 44 mo, BTFS, CSS, and MFS were not significantly different between the two groups (log rank test; BTFS: p=0.86; CSS: p=0.2; MFS: p=0.124). When matched for pT3 and high-grade tumours, CSS and MFS were significantly different between the two groups in favour of ONU (p=0.039 and p=0.004, respectively, for pT3 tumours; p=0.078 and p=0.014, respectively, for high-grade tumours). The limitations of our study include the small sample size, the single-centre experience, the personal choice of laparoscopic technique, and not performing lymphadenectomies. Perioperative data and preliminary oncologic results were presented at 22nd Congress of the European Association of Urology, Berlin, Germany. CONCLUSIONS: In patients with organ-confined UUT UCs, LNU has the advantages of minimal invasiveness and oncologic outcomes comparable to those of ONU, while its effectiveness in patients with advanced stage diseases remains to be proven. PMID- 19560260 TI - Periurethral suspension stitch during robot-assisted laparoscopic radical prostatectomy: description of the technique and continence outcomes. AB - BACKGROUND: Several studies have shown that robot-assisted laparoscopic radical prostatectomy (RALP) is feasible, with favorable complication rates and short hospital times. However, the early recovery of urinary continence remains a challenge to be overcome. OBJECTIVE: We describe our technique of periurethral retropubic suspension stitch during RALP and report its impact on early recovery of urinary continence. DESIGN, SETTING, AND PARTICIPANTS: We analyze prospectively 331 consecutive patients who underwent RALP, 94 without the placement of suspension stitch (group 1) and 237 with the application of the suspension stitch (group 2). SURGICAL PROCEDURE: The only difference between the groups was the placement of the puboperiurethral stitch after the ligation of the dorsal venous complex (DVC). The periurethral retropubic stitch was placed using a 12-in monofilament polyglytone suture on a CT-1 needle. The stitch was passed from right to left between the urethra and DVC, and then through the periostium on the pubic bone. The stitch was passed again through the DVC, and then through the pubic bone in a figure eight, and then tied. MEASUREMENTS: Continence rates were assessed with a self-administered validated questionnaire (Expanded Prostate Cancer Index Composite [EPIC]) at 1, 3, 6, and 12 mo after the procedure. Continence was defined as the use of no absorbent pads or no leakage of urine. RESULTS AND LIMITATIONS: In group 1, the continence rate at 1, 3, 6, and 12 mo postoperatively was 33%, 83%, 94.7%, and 95.7%, respectively; in group 2, the continence rate was 40%, 92.8%, 97.9%, and 97.9%, respectively. The suspension technique resulted in significantly greater continence rates at 3 mo after RALP (p=0.013). The median/mean interval to recovery of continence was also statistically significantly shorter in the suspension group (median: 6 wk; mean: 7.338 wk; 95% confidence interval [CI]: 6.387-8.288) compared to the nonsuspension group (median: 7 wk; mean: 9.585 wk; 95% CI: 7.558-11.612; log rank test, p=0.02). CONCLUSIONS: The suspension stitch during RALP resulted in a statistically significantly shorter interval to recovery of continence and higher continence rates at 3 mo after the procedure. PMID- 19560261 TI - Mechanisms of pelvic floor muscle function and the effect on the urethra during a cough. AB - BACKGROUND: Current measurement tools have difficulty identifying the automatic physiologic processes maintaining continence, and many questions still remain about pelvic floor muscle (PFM) function during automatic events. OBJECTIVE: To perform a feasibility study to characterise the displacement, velocity, and acceleration of the PFM and the urethra during a cough. DESIGN, SETTING, AND PARTICIPANTS: A volunteer convenience sample of 23 continent women and 9 women with stress urinary incontinence (SUI) from the general community of San Francisco Bay Area was studied. MEASUREMENTS: Methods included perineal ultrasound imaging, motion tracking of the urogenital structures, and digital vaginal examination. Statistical analysis used one-tailed unpaired student t tests, and Welch's correction was applied when variances were unequal. RESULTS AND LIMITATIONS: The cough reflex activated the PFM of continent women to compress the urogenital structures towards the pubic symphysis, which was absent in women with SUI. The maximum accelerations that acted on the PFM during a cough were generally more similar than the velocities and displacements. The urethras of women with SUI were exposed to uncontrolled transverse acceleration and were displaced more than twice as far (p=0.0002), with almost twice the velocity (p=0.0015) of the urethras of continent women. Caution regarding the generalisability of this study is warranted due to the small number of women in the SUI group and the significant difference in parity between groups. CONCLUSIONS: During a cough, normal PFM function produces timely compression of the pelvic floor and additional external support to the urethra, reducing displacement, velocity, and acceleration. In women with SUI, who have weaker urethral attachments, this shortening contraction does not occur; consequently, the urethras of women with SUI move further and faster for a longer duration. PMID- 19560262 TI - Interobserver variability in the detection of cerebral venous thrombosis using CT venography with matched mask bone elimination. AB - OBJECTIVES: Computed tomography venography (CTV) has proven to be a reliable imaging method in the evaluation of cerebral venous thrombosis with good correlation to magnetic resonance (MR) imaging and digital subtraction angiography (DSA). It is fast and widely accessible, especially in the emergency setting. For better visualization of vascular structures bone is often removed from the images. The purpose of this study was to evaluate the quality of a fully automatic bone removal method, matched mask bone elimination (MMBE), and to assess the interobserver variability of the CTV technique. PATIENTS AND METHODS: Fifty patients with clinical suspicion of cerebral venous thrombosis underwent multislice CTV with MMBE post-processing. Axial source images and maximum intensity projections were retrospectively evaluated by two neuroradiologists for quality of bone removal and for the presence or absence of thrombosis in nine dural sinuses and five deep cerebral veins. A per sinus/vein and a per patient analysis (thrombosis in at least one sinus or vein) was performed and interobserver agreement was assessed. RESULTS: Both observers considered bone removal good in all patients (100%). Interobserver agreement per patient was excellent (kappa=0.83), with a full agreement in 47 of 50 patients (94%). The interobserver agreement per sinus or vein was good (kappa=0.76), with a full agreement in 679 of 700 sinuses or veins (97%). CONCLUSION: CTV aided with MMBE is a robust technique for visualization of the intracranial venous circulation, removing bone effectively. CTV has high interobserver agreement for presence or absence of cerebral venous thrombosis. PMID- 19560263 TI - A small molecule inhibitor of NF-kappaB, dehydroxymethylepoxyquinomicin (DHMEQ), suppresses growth and invasion of nasopharyngeal carcinoma (NPC) cells. AB - Despite the demonstrated constitutive activation of NF-kappaB in nasopharyngeal carcinoma (NPC), the therapeutic potential of targeting this pathway has not been investigated. Here, we employed a small molecule inhibitor of NF-kappaB, DHMEQ (which mainly blocks nuclear translocation of activated NF-kappaB) and demonstrated significant inhibition of NPC cell proliferation, migration, invasion, as well as anchorage-independent growth. These antitumor effects were associated with induction of G(2)/M cell cycle arrest and apoptosis, and downregulation of NF-kappaB target genes (EGFR, cyclin D1 and survivin). This first demonstration of therapeutic benefits of NF-kappaB targeting in NPC implicates the importance of targeting this pathway in NPC. PMID- 19560264 TI - Rapamycin increases the p53/MDM2 protein ratio and p53-dependent apoptosis by translational inhibition of mdm2 in cancer cells. AB - Rapamycin, a potential anti-cancer agent, modulates activity of various factors functioning in translation, including eIF4E, an initiation factor selectively regulating expression of a subset of cellular transcripts. We show here that rapamycin suppresses levels of the p53-regulator MDM2 by translational inhibition without affecting mdm2 mRNA expression or protein stability. Rapamycin inhibits translation of mdm2 mRNA from the constitutive P1 promoter, which contains two upstream ORFs (uORFs) in the 5'UTR. Suppression is accompanied by increased hypo phosphorylation of 4EBP-1, an inhibitory eIF4E binding protein. Ectopic expression of eIF4E abrogates rapamycin-mediated MDM2 inhibition, suggesting that eIF4E is crucial in modulating MDM2 expression in rapamycin-treated cells. Rapamycin administration also results in elevated PUMA expression and PARP cleavage, which is reproduced by siRNA knockdown of eIF4E or MDM2, suggesting that MDM2 suppression by rapamycin stimulates p53-mediated apoptosis. Together, our results define translational regulation of MDM2 expression by eIF4E and provide a molecular mechanism underlying rapamycin-induced p53-dependent apoptosis. PMID- 19560265 TI - The investigation of catalytic ozonation and integrated catalytic ozonation/biological processes for the removal of phenol from saline wastewaters. AB - The effectiveness of the catalytic ozonation process (COP) with a GAC catalyst was assessed based on the degradation and COD removal of phenol from the saline wastewater, as compared with the single ozonation process (SOP). The COP attained a much higher level of phenol degradation compared to the SOP. The influence of several variables was investigated, including pH of solution, NaCl concentration, and dosage of GAC, for their effects on COP phenol degradation in a synthetic saline wastewater. The maximum degradation of phenol was achieved at pH 8 and 20 g/L GAC. NaCl had no adverse effect on phenol removal at ranges between 0.5 and 50 g/L. The activated carbon acted mostly as a catalyst for ozone decomposition, and the subsequent generation of hydroxyl radicals. Furthermore, the GAC preserved its catalytic properties after 5 times reuse. The capability of a biological process to treat COP effluent was also investigated. Results showed that a 10 min reaction time in COP under optimum conditions reduces the concentrations of phenol and COD to an acceptable level for efficient post treating in a suspended growth bioreactor at a short aeration time of 4h. Thus, the integration of COP with a biological process is proven to be a technically and economically effective method for treating saline wastewaters containing recalcitrant compounds. PMID- 19560267 TI - Application of natural zeolite for phosphorus and ammonium removal from aqueous solutions. AB - Removal of both nutrients ammonium and phosphorus by natural zeolite has been studied in lab scale by using a mechanically stirred batch system (1000 ml). Zeolite, a mean particle size of 13 microm, was used as an adsorbent for the removal of ammonium and then as a seed material for the precipitation of calcium phosphate. A relationship was established between the uptake of ammonium by zeolite and the ratio of initial ammonium concentration to zeolite dosage. Ammonium uptake of zeolite was almost completed within initial 5 min of adsorption period. There is no pronounced effect of zeolite and ammonium, neither positive nor negative on the amount of calcium phosphate precipitation. The extent of the precipitation of phosphate increased with rising pH. It was also observed that when the system was allowed to relax at constant pH (i.e. under relatively low super saturations), a certain lag time was noted to elapse at the onset of the precipitation. At the pH 7.2, the amount of initial fast precipitation within 5 min and total precipitation within 120 min were around 34% and 93%, respectively. Precipitation of calcium phosphate on to ammonium-loaded zeolite was achieved at low super saturations (< pH 7.5) through secondary nucleation and crystal growth, leading to an increase in particle size. PMID- 19560266 TI - Electrochemical determination of cadmium(II) at platinum electrode modified with kaolin by square wave voltammetry. AB - In this work, determination of cadmium(II) using square wave voltammetry (SWV) was described. The method is based on accumulation of these metal ions on kaolin platinum electrode (K/Pt). The K/Pt performance was optimized with respect to the surface modification and operating conditions. The optimized conditions were obtained in pH of 5.0 and accumulation time of 25 min. Under the optimal conditions, the relationship between the peak current versus concentration was linear over the range of 9 x 10(-8) to 8.3 x 10(-6) mol L(-1). The detection limit (DL, 3sigma) was 5.4 x 10(-9) mol L(-1). The analytical methodology was successfully applied to monitor the Cd(II) content in natural water. Interferences were also evaluated. PMID- 19560268 TI - Elaboration, characterization and application of polysulfone and polyacrylic acid blends as ultrafiltration membranes for removal of some heavy metals from water. AB - Polysulfone (PSf)/polyacrylic acid ultrafiltration (PSf/PAA) membranes were prepared from a polymer blend in dimethylformamide by coagulation in water according to the wet phase inversion method. Immobilization of water-soluble PAA within the non-soluble PSf matrix was proven by the increase of ion exchange capacity and the intensity of the carboxyl groups' peak with the increase of PAA content as shown by Fourier transform infrared spectra. These results lead to consider that PSf and PAA form a semi-interpenetrating polymer networks. The obtained membranes showed a decrease of mean surface-pore sizes, the overall porosity and the hydraulic permeability with the increase in PAA content. Such results were imputed to the morphologic modifications of PSf film with the immobilization of increasing PAA amount. PSf/PAA membranes showed high lead, cadmium and chromium rejection which reaches 100% at pH superior to 5.7 and a low rejection at low pH. Moreover, the heavy metal rejection decreases with feed solution concentration and applied pressure increases. These behaviors were attributed to the role of carboxylic groups in ion exchange or complexation. As a matter of fact, the strong lead ion-PAA interactions were revealed by the scanning electron microscopy with energy dispersive X-rays (SEM-EDX). PMID- 19560269 TI - Effect of the adsorbate (Bromacil) equilibrium concentration in water on its adsorption on powdered activated carbon. Part 2: Kinetic parameters. AB - The application of several monosolute equilibrium models has previously shown that Bromacil adsorption on SA-UF (Norit) powdered activated carbon (PAC) is probably effective on two types of sites. High reactivity sites were found to be 10-20 less present in a carbon surface than lower reactivity sites, according to the q(m) values calculated by isotherm models. The aims of this work were trying, primarily, to identify the kinetic-determinant stage of the sorption of Bromacil at a wide range of initial pesticide concentrations (approximately 5 to approximately 500 microg L(-1) at pH 7.8), and secondly, to specify the rate constants and other useful design parameters for the application in water treatment. It was therefore not possible to specify a priori whether the diffusion or surface reaction is the key step. It shows that many of the tested models which describe the stage of distribution or the surface reaction are correctly applied. However, the diffusivity values (D and D(0)) were found to be constant only constants for some specific experimental concentrations. The HSDM model of surface diffusion in pores was also applied but the values of the diffusion coefficient of surface (D(s)) were widely scattered and reduce significantly with the initial concentration or the equilibrium concentration in Bromacil. The model of surface reaction of pseudo-second order fitted particularly well and led to constant values which are independent of the equilibrium concentration, except for the low concentrations where the constants become significantly more important. This last observation confirms perfectly the hypothesis based on two types of sites as concluded by the equilibrium data (part 1). PMID- 19560271 TI - fMRI investigation of the effect of local and systemic lidocaine on noxious electrical stimulation-induced activation in spinal cord. AB - Spinal cord fMRI offers an excellent opportunity to quantify nociception using neuronal activation induced by painful stimuli. Measurement of the magnitude of stimulation-induced activation, and its suppression with analgesics can provide objective measures of pain and efficacy of analgesics. This study investigates the feasibility of using spinal cord fMRI in anesthetized rats as a pain assay to test the analgesic effect of locally and systemically administered lidocaine. Blood volume (BV)-weighted fMRI signal acquired after intravenous injection of ultrasmall superparamagnetic iron oxide (USPIO) particles was used as an indirect readout of the neuronal activity. Transcutaneous noxious electrical stimulation was used as the pain model. BV-weighted fMRI signal could be robustly quantified on a run-by-run basis, opening the possibility of measuring pharmacodynamics (PD) of the analgesics with a temporal resolution of approximately 2 min. Local administration of lidocaine was shown to ablate all stimulation-induced fMRI signals by the total blockage of peripheral nerve transmission, while the analgesic effect of systemically administered lidocaine was robustly detected after intravenous infusion of approximately 3mg/kg, which is similar to clinical dosage for human. This study establishes spinal cord fMRI as a viable assay for analgesics. With respect to the mode of action of lidocaine, this study suggests that systemic lidocaine, which is clinically used for the treatment of neuropathic pain, and believed to only block the peripheral nerve transmission of abnormal neural activity (ectopic discharge) originating from the damaged peripheral nerves, also blocks the peripheral nerve transmission of normal neural activity induced by transcutaneous noxious electrical stimulation. PMID- 19560270 TI - Sex differences in functional brain activation during noxious visceral stimulation in rats. AB - Studies in healthy human subjects and patients with irritable bowel syndrome suggest sex differences in cerebral nociceptive processing. Here we examine sex differences in functional brain activation in the rat during colorectal distention (CRD), a preclinical model of acute visceral pain. [(14)C] iodoantipyrine was injected intravenously in awake, non-restrained female rats during 60- or 0-mmHg CRD while electromyographic abdominal activity (EMG) and pain behavior were recorded. Regional cerebral blood flow-related tissue radioactivity was analyzed by statistical parametric mapping from autoradiographic images of three-dimensionally reconstructed brains. Sex differences were addressed by comparing the current data with our previously published data collected from male rats. While sex differences in EMG and pain scores were modest, significant differences were noted in functional brain activation. Females showed widespread changes in limbic (amygdala, hypothalamus) and paralimbic structures (ventral striatum, nucleus accumbens, raphe), while males demonstrated broad cortical changes. Sex differences were apparent in the homeostatic afferent network (parabrachial nucleus, thalamus, insular and dorsal anterior cingulate cortices), in an emotional-arousal network (amygdala, locus coeruleus complex), and in cortical areas modulating these networks (prefrontal cortex). Greater activation of the ventromedial prefrontal cortex and broader limbic/paralimbic changes in females suggest greater engagement of affective mechanisms during visceral pain. Greater cortical activation in males is consistent with the concept of greater cortical inhibitory effects on limbic structures in males, which may relate to differences in attentional and cognitive attribution to visceral stimuli. These findings show remarkable similarities to reported sex differences in brain responses to visceral stimuli in humans. PMID- 19560272 TI - Experimental arthritis causes tumor necrosis factor-alpha-dependent infiltration of macrophages into rat dorsal root ganglia which correlates with pain-related behavior. AB - After peripheral nerve damage macrophages infiltrate the dorsal root ganglia (DRG) in which cell bodies of lesioned neurons are located. However, infiltration of macrophages into the DRGs was also reported in complete Freund's adjuvant (CFA)-induced inflammation raising the question whether CFA inflammation induces nerve cell damage or whether peripheral inflammation may also trigger macrophage infiltration into DRGs. Related questions are, first, which signals trigger macrophage infiltration into DRGs and, second, is macrophage infiltration correlated with pain-related behavior. Using the rat model of unilateral antigen induced arthritis (AIA) in the knee we found a massive infiltration of ED1(+) macrophages into the ipsi- and contralateral lumbar DRGs but not into thoracic DRGs. At no time point of AIA DRG neurons showed expression of activating transcription factor-3 (ATF3) indicating that macrophage infiltration is not explainable by nerve cell lesions in this model. During AIA, lumbar but not thoracic DRGs exhibited a bilateral de novo expression of vascular cell adhesion molecule-1 (VCAM-1) which is known to be involved in macrophage infiltration. Tumor necrosis factor-alpha (TNF-alpha) neutralization with etanercept or infliximab treatment after induction of AIA significantly reduced both macrophage infiltration and VCAM-1 expression. It also decreased mechanical hyperalgesia at the inflamed joint although the joint inflammation itself was barely attenuated, and it reduced mechanical hyperalgesia at the non-inflamed contralateral knee joint. Thus, bilateral segment-specific infiltration of macrophages into DRGs is part of an unilateral inflammatory process in peripheral tissue and it may be involved in the generation of hyperalgesia in particular on the non-inflamed side. PMID- 19560273 TI - Comparative immunogenicity of Haemaphysalis longicornis and Rhipicephalus (Boophilus) microplus calreticulins. AB - The ticks Rhipicephalus (Boophilus) microplus and Haemaphysalis longicornis are blood-sucking ectoparasites of bovines, causing serious damages to the livestock production. The main control method for these ticks is based on acaricides. However, the use of vaccines has been studied as a promising control strategy. Calreticulin (CRT) is a multifunctional, predominantly intracellular protein present in almost all cells of animals. The secretion of CRT during feeding might be linked to the modulation of the parasite-host interaction. In the present study, recombinant CRTs of R. microplus (rBmCRT) and H. longicornis (rHlCRT) were expressed in Escherichia coli and purified by ion exchange chromatography and used for the immunization of bovines and mouse. ELISA demonstrated that both rCRTs are recognized by the sera of immunized bovines. In silico, despite the difference in amino acid sequences, antigenic index analysis of HlCRT and BmCRT using the Jameson-Wolf algorithm indicated that both proteins were very similar in antigenicity index, although six different epitopes between the tick CRTs have been inferred. These data were corroborated by competitive ELISA analyses, which suggest the presence of different epitopes within the proteins. Western blot analyses showed that anti-rBmCRT and anti-rHlCRT bovine sera also recognized the native proteins in larvae extracts and, moreover, sera of bovines immunized with saliva and extract of salivary glands recognized both recombinant CRTs. Thus, mouse and bovine immune system recognized rCRTs, resulting in the production of antibodies with similar specificity for both recombinant proteins, although different epitopes could be distinguished between rBmCRT and rHlCRT. PMID- 19560274 TI - The Arctic as a model for anticipating, preventing, and mitigating climate change impacts on host-parasite interactions. AB - Climate change is influencing the structure and function of natural ecosystems around the world, including host-parasite interactions and disease emergence. Understanding the influence of climate change on infectious disease at temperate and tropical latitudes can be challenging because of numerous complicating biological, social, and political factors. Arctic and Subarctic regions may be particularly good models for unraveling the impacts of climate change on parasite ecology because they are relatively simple systems with low biological diversity and few other complicating anthropogenic factors. We examine some changing dynamics of host-parasite interactions at high latitudes and use these to illustrate a framework for approaching understanding, preventing, and mitigating climate change impacts on infectious disease, including zoonoses, in wildlife. PMID- 19560275 TI - Influence of climatic factors on dynamics of questing Ixodes ricinus ticks in Slovenia. AB - Ixodes ricinus is a vector of pathogens that cause many diseases in Europe and Slovenia: tick-borne encephalitis (TBE), anaplasmosis, borreliosis, babesiosis and others. The risk for contracting these diseases depends strongly on the density of the infected questing ticks and many studies have investigated tick population dynamics and the parameters affecting them. They have shown a clear influence of climatic and landscape arrangements in the microhabitat on tick abundance and dynamics and therefore on transmission of pathogens important in human and veterinary medicine. In our study we assessed the influence of climatic factors on questing activity of ticks over a three-year period at 7 locations in Slovenia. Locations were selected in endemic foci of TBE with different intensity, which were identified according to the presence of human disease. Sites differ according to various abiotic and biotic factors, such as climate, amount of rain, height above sea level, vegetation and wildlife. All three stages of ticks were collected monthly over a three-year period (2005-2007). Temperature, humidity and precipitation data were collected for these years. The purpose of our study was to relate observed differences in I. ricinus ticks questing activity to local climate. We found a correlation between the decrease of questing ticks in the summer and the combination of air temperatures and humidity in the form of saturation deficit. PMID- 19560277 TI - From parasite genomes to one healthy world: Are we having fun yet? AB - In 1990, the Human Genome Sequencing Project was established. This laid the ground work for an explosion of sequence data that has since followed. As a result of this effort, the first complete genome of an animal, Caenorhabditis elegans was published in 1998. The sequence of Drosophila melanogaster was made available in March, 2000 and in the following year, working drafts of the human genome were generated with the completed sequence (92%) being released in 2003. Recent advancements and next-generation technologies have made sequencing common place and have infiltrated every aspect of biological research, including parasitology. To date, sequencing of 32 apicomplexa and 24 nematode genomes are either in progress or near completion, and over 600k nematode EST and 200k apicomplexa EST submissions fill the databases. However, the winds have shifted and efforts are now refocusing on how best to store, mine and apply these data to problem solving. Herein we tend not to summarize existing X-omics datasets or present new technological advances that promise future benefits. Rather, the information to follow condenses up-to-date-applications of existing technologies to problem solving as it relates to parasite research. Advancements in non parasite systems are also presented with the proviso that applications to parasite research are in the making. PMID- 19560276 TI - Comparative assessment of albendazole and triclabendazole ovicidal activity on Fasciola hepatica eggs. AB - The benzimidazole compounds albendazole (ABZ) and triclabendazole (TCBZ) are both effective against Fasciola hepatica, although ABZ is only effective against adult flukes. Additionally, ABZ is a broad-spectrum nematodicidal compound with well known ovicidal activity. However, no data on the ovicidal effect of TCBZ against F. hepatica eggs are available. The work reported here evaluated the comparative ovicidal effect of ABZ, TCBZ and their sulphoxide metabolites on F. hepatica eggs recovered from bile of sheep artificially infected with either a TCBZ-susceptible (Cullompton) or a TCBZ-resistant (Sligo) isolate of F. hepatica. Additionally, the effects of different non-flukicidal methylcarbamate benzimidazole compounds on the hatching of F. hepatica eggs were evaluated. Eggs (500 eggs/mL, n=4) were incubated for 12h either with TCBZ, TCBZ sulphoxide (TCBZ.SO), ABZ (5, 10 and 20 nmol/mL) or without drug (untreated control) (Experiment 1). Additionally, the effect of TCBZ and TCBZ.SO (5 nmol/mL) on egg hatchability was examined after a long (15 days) drug exposure (Experiment 2). Furthermore, the ovicidal effect of ABZ and ABZ.SO at different concentrations (5, 1, 0.5, 0.1 and 0.05 nmol/mL) (Experiment 3), and the effect of fenbendazole (FBZ), oxfendazole (OFZ), mebendazole (MBZ), flubendazole (FLBZ) (5 nmol/mL) and reduced-FLBZ (R-FLBZ) (2 microg/mL) on fluke eggs, were evaluated after a 12-h exposure (Experiment 4). Egg hatch was assessed by direct microscopic observation after incubation at 25 degrees C for 15 days. TCBZ and TCBZ.SO did not affect egg hatch after a 12-h incubation. A similar result was obtained after a much longer drug exposure (15 days) (Experiment 1 and 2). However, a significant (P<0.05) inhibition of egg hatch was observed in ABZ- and ABZ.SO-incubated eggs (Experiments 1 and 3). Additionally, the non-flukicidal compounds (Experiment 4) affected egg hatchability, particularly FLBZ and R-FLBZ. In conclusion, ABZ and ABZ.SO had a clear inhibitory effect on egg development of F. hepatica. However, the most extensively used flukicidal compound, TCBZ, and its main sulphoxide metabolite, did not affect egg hatch, even in TCBZ-susceptible flukes. PMID- 19560278 TI - Stress hormones mediate drug resistance to paclitaxel in human breast cancer cells through a CDK-1-dependent pathway. AB - Chemotherapy comprises part of successful treatment regimens for breast cancer, however, up to 50% of patients develop resistance. Stress in cancer patients can equate to poor chemotherapeutic responses. We hypothesize that drug resistance may be associated with stress hormone-induced alterations in breast cancer cells. To test this hypothesis, MDA-MB-231 cells were cultured with paclitaxel and/or cortisol, norepinephrine and epinephrine and cytotoxicity, cell cycle analyses, genomic and proteomic analyses were performed. Paclitaxel-mediated cytotoxicity and G2/M cell cycle arrest were reversed significantly by stress hormones. Genomic and proteomic analyses revealed that stress hormones modulated beta tubulin isotypes and significantly altered genes and proteins involved in regulation of the G2/M transition, including cyclin-dependent kinase-1 (CDK-1). Inhibition of CDK-1 abrogated stress hormone-mediated reversal of paclitaxel induced cytotoxicity, indicating that the protective effect of stress hormones act through a CDK-1-dependent mechanism. These data demonstrate that stress hormones interfere with paclitaxel efficacy and contribute significantly to drug resistance. PMID- 19560279 TI - The role of FKBP5, a co-chaperone of the glucocorticoid receptor in the pathogenesis and therapy of affective and anxiety disorders. AB - FK506 binding protein 51 or FKBP5 is a co-chaperone of hsp90 which regulates glucocorticoid receptor (GR) sensitivity. When it is bound to the receptor complex, cortisol binds with lower affinity and nuclear translocation of the receptor is less efficient. FKBP5 mRNA and protein expression are induced by GR activation via intronic hormone response elements and this provides an ultra short feedback loop for GR-sensitivity. Polymorphisms in the gene encoding this co-chaperone have been shown to associate with differential upregulation of FKBP5 following GR activation and differences in GR sensitivity and stress hormone system regulation. Alleles associated with enhanced expression of FKBP5 following GR activation, lead to an increased GR resistance and decreased efficiency of the negative feedback of the stress hormone axis in healthy controls. This results in a prolongation of stress hormone system activation following exposure to stress. This dysregulated stress response might be a risk factor for stress-related psychiatric disorders. In fact, the same alleles are over-represented in individuals with major depression, bipolar disorder and post-traumatic stress disorder. In addition, they are also associated with faster response to antidepressant treatment. FKBP5 might thus be an interesting therapeutic target for the prevention and treatment of stress-related psychiatric disorders. PMID- 19560281 TI - Early orthodontic treatment of skeletal Class II malocclusion may be effective to prevent the potential for OSAHS and snoring. AB - OSAHS or snoring is an important condition within our community with the potential of being a significant health burden. Although the precise pathogenesis of upper airway obstruction during sleep remains uncertain in OSAHS and snoring patients, craniofacial risk factors are said to be associated with OSAHS and snoring. Since a high number of OSAHS and snoring patients consist of skeletal Class II malocclusion patients characterized by deficient mandible, then we can make the hypotheses that early orthodontic treatment of skeletal Class II malocclusion patients to improve such discrepancies during the growth period may be effective to prevent the potential for OSAHS and snoring. PMID- 19560280 TI - Perceived support to stay quit: what happens after delivery? AB - OBJECTIVE: To assess whether perceived changes in postpartum support were associated with postpartum return to smoking. STUDY DESIGN: This is a prospective repeated measures, mixed methods observational study. Sixty-five women who smoked prior to pregnancy were recruited at delivery and surveyed at 2, 6, 12, and 24 weeks postpartum; in-depth interviews were conducted when participants reported smoking. RESULTS: Fifty-two percent self identified as White, non-Hispanic. Forty seven percent resumed smoking by 24 weeks postpartum. Women who had returned to smoking by 24 weeks had a significantly larger decrease in perceived smoking specific support than women who remained abstinent (p<0.001). By 24-week postpartum follow-up, only 24% of women reported that an obstetric clinician had discussed how to quit/stay quit. When qualitatively interviewed, more than half of women reported having no one to support them to stay quit or quit smoking. CONCLUSION: Following delivery, women lack needed smoking-specific support. Decline in perceived smoking-specific support from family and friends is associated with postpartum smoking resumption. PMID- 19560282 TI - Epigenetic regulation of glycosylation could be a mechanism used by complex organisms to compete with microbes on an evolutionary scale. AB - Glycosylation is the most diverse post-translational protein modification. It is essential for multicellular life and its complete absence is embryonically lethal. Hundreds of specific enzymes are involved in the synthesis of complex oligosaccharide structures that are covalently bound to protein backbones. This process is not template driven and thus results in a huge complexity of glycoproteome, estimated to be several orders of magnitude larger than proteome. Large structural variability provided by glycans represents a significant evolutionary advantage and nearly all proteins invented after the appearance of the multicellular life are glycosylated. Glycosylation represents a way how complex organisms could develop novel structural features without introducing probably deleterious changes in their genome. Intricate mechanisms by which the interplay of gene expression and intracellular localization of their products give rise to specific glycan structures is only starting to be understood, but some evidence suggests that epigenetic regulation of glycosylation might be used to create novel biological structures. Here we suggest a hypothesis that epigenetic regulation of genes involved in glycan synthesis might represent a way how newly developed structural advantages could be transmitted through generations, thus providing a tool for complex organisms to compete with high speed of evolution of unicellular organisms. PMID- 19560283 TI - Omental milky spots--highly efficient "natural filter" for screening gastric cancer stem cells. AB - Over the last decades, cancer stem cells (CSCs) have been identified in different solid tumors and three strategies have been employed to identify them as follows: (1) expression of cell surface markers, (2) ability to exclude fluorescent dye Hoescht 33342 in a cell population termed "side population" (SP), and (3) formation of tumorspheres in suspension culture. But these methods focus on a minor subset of tumor cells, initial CSCs are still hidden among them and genuine CSCs have not yet been isolated and purified. Here we propose a new strategy to identify and isolate gastric cancer stem cells--using omental milky spots to screen gastric cancer stem cells in peritoneal metastasis mouse models of gastric cancer. This strategy directly target cancer stem cells--which can cause a new tumor by itself. Omental milky spots and their macrophages become a highly efficient "natural filter" for screening gastric cancer stem cells. PMID- 19560284 TI - Impeded venous drainage: novel view of chronic venous disease pathophysiology. AB - Venous valve incompetence role as the primary pathology occurring in patients with chronic venous diseases (CVD) has long been exaggerated. Management strategies based on the current pathophysiological concepts are far from perfect. A novel concept is introduced describing the importance of patency of axial venous system with "Pressure gradient model" providing the venous system architecture required for efficient venous drainage. The roles of calf muscle pump, perforators and venous valves according to the novel concept are described. Impeded venous drainage (IVD) plays the key role in the pathophysiology of CVD. Management of CVD should be directed to correction of IVD instead of correction or ablation of valve incompetence. PMID- 19560285 TI - The mainstream hypothesis that LDL cholesterol drives atherosclerosis may have been falsified by non-invasive imaging of coronary artery plaque burden and progression. AB - That LDL cholesterol drives atherosclerosis is a widely if not almost universally held belief, and this belief strongly influences the mainstream approach to coronary heart disease. However heart disease has a number of stages, and in terms of primary prevention, the initiation and progression of silent or sub clinical atherosclerosis is clearly fundamental. However, studies that address the efficacy of interventions and practices aimed at the primary prevention of heart disease almost always use event-based endpoints such as fatal or non-fatal myocardial infarction or unstable angina. These endpoints do not directly relate to the primary prevention of silent atherosclerosis and to apply these results to asymptomatic individuals in this context involves an extrapolation. The advent of non-invasive imaging techniques which allow the determination of coronary artery plaque burden and progression of plaque has provided a unique opportunity to examine the relationship between both traditional and emerging risk factors and the extent of sub-clinical coronary artery disease and in particular allow the testing of the hypothesis that LDL cholesterol drives coronary atherosclerosis. Consistent with earlier autopsy studies, the use of electron beam tomography and contrast enhanced CT angiography techniques have created a large body of evidence which appears to falsify this hypothesis. The large number of null results for the association between serum LDL cholesterol levels and the prevalence or progression of both calcified and non-calcified plaque in the appropriate vascular bed and involving large numbers of men and women over a wide range of age, ethnic background, plaque burden and cholesterol levels cannot be easily dismissed. If the hypothesis is false, this has a significant impact on currently held views regarding risk factors and therapeutic interventions in the case of individuals who are asymptomatic, that is, issues associated with primary prevention. Also, if the hypothesis is false, then the use of changes in LDL as a surrogate marker for judging the importance of various risk factors for silent atherosclerosis and thus coronary artery disease can be called into question. PMID- 19560286 TI - Role of corticotropin-releasing hormone and receptor in the pathogenesis of psoriasis. AB - Psoriasis is a chronic inflammatory disease characterized by epidermal keratinocytic hyperproliferation and abnormal differentiation. It is one of the most illustrative examples of the close relation between exacerbation of disease and the psychopathologic burden of the patients. However, the mechanism remains poorly understood. In recent years, evidence has suggested that endocrine stress responses not only are under control of the central nervous system but also occur in peripheral tissue, outside of the classical HPA axis. Corticotrophin-releasing hormone (CRH) is a central component of the hypothalamic-pituitary-adrenal (HPA) axis and is an important coordinator of the systemic stress response. In peripheral sites, cutaneous CRH and CRH-receptor1 (CRH-R1) is believed to regulate various functions of the skin that are important for local homeostasis. These findings have shed new light on the role of peripheral CRH and CRH-R1 in cutaneous diseases, especially psoriasis. Many researchers focus on the pro inflammatory role of CRH, such as CRH-induced activation of mast cells in the phenomenon of stress related exacerbation of cutaneous inflammatory diseases, and some researches demonstrated that CRH protein expression was increased in the affected skin of psoriasis. Meanwhile, it is reported that CRH could downregulate pro-inflammatory factors, such as IL-18. Tagen found CRH-R1 mRNA expression in psoriasis skin lower than that in normal controls. Previous studies revealed that the functional role of the CRH/CRH-R1 system in pathological human skin conditions remains to be identified. Interestingly, we found that both CRH and CRH-R1 were presented in psoriatic lesion, perilesional skin and normal control skin by immunohistochemistry, and lesions from patients with psoriasis showed significantly lower CRH/CRH-R1 expression compared with psoriatic perilesional skin and normal control skin. Presumably a localized circuit regulates the peripheral functions of cutaneous CRH/CRH-R1, and the aberrant expression of CRH/CRH-R1 in the skin disturbs the local homeostasis and leads to abnormal differentiation and proliferation in keratinocytes. However, dysfunction of keratinocytes may decrease CRH/CRH-R1 expression because of disharmony in differentiation and proliferation of keratinocytes. Thus, we hypothesize that a cutaneous CRH/CRH-R1 system might be aberrant in lesions of psoriasis. The detuning of CRH/CRH-R1 regulation might contribute to the formation of plaque in psoriasis. What is more important, we hypothesize that the role of CRH/CRH-R1 system might play a protective role in the pathogenesis of psoriasis. This would provide a new treatment for psoriasis. Thus, further study in vitro and in vivo has to be done to test our hypothesis. PMID- 19560287 TI - In vivo self-expanding engineering of bone. AB - In vivo bone formation and vascularization are interdependent process with complex regulatory mechanism. In vitro tissue engineering has meet great difficulty to copy all internal environment conditions, and the technology worked well only for relatively simple, thin tissues such as skin and cartilage that do not depend on a well-formed network of blood vessels to deliver food and oxygen. For more complex tissue of bone segments shot through with vessels, we hypothesize that an altered strategy might be effective: starting from in vitro tissue engineering for a thin sheet of primary product, then transferring to in vivo cultivation with a self-expanding procedure activated by interweaving metal springs of shape memory alloy (SMA). It is easy for thin sheet of engineering tissues to survive after implantation in vivo. Super-elasticity of SMA spring can gradually expand and re-shape the engineering product. More important, the continuous mechano-transduction effect can activate internal environment as a bioreactor for bone regeneration and vascularization. Our hypothesis integrates the existing knowledge and technologies, and could be a hopeful strategy for more convenient and safe use of bone engineering in clinics. PMID- 19560288 TI - Platelets might mediate the increase of plasma glutamate in acute ischemic stroke: relevance for early neurological deterioration. AB - In this brief paper we would like to hypothesize that platelets might represent a "peripheral" contributory source for the elevation of plasma glutamate levels in the setting of acute ischemic stroke: available evidence and possible mechanisms will be discussed, especially drawing attention to the possible relevance for the pathophysiology of early neurological deterioration. PMID- 19560289 TI - Cortistatin vaccination--a solution to growth hormone deficiency. AB - Cortistatin and somatostatin are neuropeptides which have inhibitory effects on growth hormone through common five receptors. Although, both have inhibitory effects but, only cortistatin has direct inhibitory effects on growth hormone secretagogue and is more potent inhibitor of growth hormone than somatostatin. This control of growth hormone can be manipulated through immunoneutralization of cortistatin through cortistatin DNA vaccine rather than antibodies application. A DNA vaccine of cortistatin can be produced using recombinant DNA technology in a eukaryotic expression system and will serve as a tool not to only alleviate the growth hormone deficiency problems in human but, can also be used to improve growth rate in farm animals. PMID- 19560290 TI - Modafinil for the treatment of cocaine dependence. AB - AIM: Modafinil was tested for efficacy in facilitating abstinence in cocaine dependent patients, compared to placebo. METHODS: This was a double-blind placebo controlled study, with 12 weeks of treatment and a 4-week follow-up. Six outpatient substance abuse treatment clinics participated in the study. There were 210 treatment-seekers randomized, having a diagnosis of cocaine dependence; 72 participants were randomized to placebo, 69 to modafinil 200mg, and 69 to modafinil 400mg, taken once daily on awakening. Participants came to the clinic three times per week for assessments and urine drug screens, and had one hour of individual psychotherapy weekly. The primary outcome measure was the weekly percentage of cocaine non-use days. RESULTS: The GEE regression analysis showed that for the total sample, there was no significant difference between either modafinil group and placebo in the change in average weekly percent of cocaine non-use days over the 12-week treatment period (p>0.79). However, two secondary outcomes showed significant effects by modafinil 200mg: the maximum number of consecutive non-use days for cocaine (p=0.02), and a reduction in craving (p=0.04). Also, a post hoc analysis showed a significant effect of modafinil that increased the weekly percentage of non-use days in the subgroup of those cocaine patients who did not have a history of alcohol dependence (p<0.02). CONCLUSIONS: These data suggest that modafinil, in combination with individual behavioral therapy, was effective for increasing cocaine non-use days in participants without co-morbid alcohol dependence, and in reducing cocaine craving. PMID- 19560291 TI - Co-isolation of Bartonella henselae and Bartonella vinsonii subsp. berkhoffii from blood, joint and subcutaneous seroma fluids from two naturally infected dogs. AB - This report describes the clinical presentation, isolation and treatment of two dogs naturally infected with Bartonella henselae and Bartonella vinsonii subsp. berkhoffii. Chronic and progressive polyarthritis was the primary complaint for dog #1, from which B. henselae and B. vinsonii subsp. berkhoffii were cultured on three independent occasions from blood and joint fluid samples, despite administration of nearly 4 months of non-consecutive antibiotic therapy. A clinically atypical and progressively severe trauma-associated seroma was the primary complaint for dog #2, from which B. henselae and B. vinsonii subsp. berkhoffii were isolated from serum, blood and seroma fluid. Dogs can be co infected with two Bartonella spp. and infection with these organisms should not be ruled out if specific antibodies are not detected. Specialized culture techniques should be used for isolation and to assess antibiotic efficacy. PMID- 19560292 TI - High prevalence of co-infections with bovine herpesvirus 1 and 5 found in cattle in southern Brazil. AB - Based on small scale studies or on little sensitive serological tests, bovines in the south of the state of Rio Grande do Sul, Brazil, are known to be infected with either bovine herpesvirus 1 (BoHV-1) or 5 (BoHV-5). However, whether the prevalence of each of these viruses is high or low is currently still unknown. In order to determine the extent of BoHV (-1 and/or -5) infections in bovines in this region of Brazil, 200 bovines were studied for the presence of BoHV DNA. To this end, first a quantitative PCR was developed that amplified BoHV-1 DNA as well as BoHV-5 DNA. Using this PCR the number of BoHV genomes normally present in latently infected ganglia of naturally infected bovines was estimated. The new PCR was sensitive enough to detect most BoHV DNA in infected ganglia. The results of this first PCR showed that at least 87% of the bovines in the south of Rio Grande do Sul were (latently) infected with either BoHV-1 or BoHV-5. To determine the prevalence of BoHV-1 and BoHV-5 separately, two type-specific PCRs - one for each virus - were developed that used the products of the first PCR as a template. The results of these type-specific PCRs showed that 82.8% of the BoHV positive population was (latently) infected with BoHV-1, 93.1% with BoHV-5 and 75.9% with both BoHV-1 and BoHV-5. This is the first time that such a high frequency of co-infection of BoHV-1 and BoHV-5 in bovines has been demonstrated. PMID- 19560293 TI - Pain and distress reactivity and recovery as early predictors of temperament in toddlers born preterm. AB - BACKGROUND: Pain reactivity may reflect underlying mechanisms of constitutional aspects of temperament. AIM: To examine whether the neonatal biobehavioral reactivity and recovery responses from pain and distress, as well as the gestational age, the illness severity and the amount of painful procedures undergone the Neonatal Intensive Care Unit (NICU) stay, predict temperament later in toddlerhood, in vulnerable children born preterm. STUDY DESIGN: Prospective longitudinal study. SUBJECTS: Twenty-six preterm and very low birth weight infants followed from birth to toddlerhood. OUTCOME MEASURES: Illness severity was assessed with the Clinical Risk Index for Babies (CRIB) score. The medical charts were reviewed prospectively for obtaining the amount of pain exposure in NICU. For assessing the behavioral and cardiac reactivity and recovery from pain and distress, the neonates were evaluated during routine blood collection in the NICU in the first 10 days of life. Pain and distress reactivity and recovery was measured using the Neonatal Facial Coding System score, the duration of crying, and the magnitude of average heart rate. At toddlerhood, mothers answered the Early Childhood Behavior Questionnaire. RESULTS: Higher biobehavioral reactivity to pain and distress predicted higher temperamental Negative Affect, above and beyond gestational age, illness severity and amount of pain exposure in NICU. However, we did not find a predictive relation between gestational age, CRIB score and number of painful procedures undergone NICU and toddler's temperament. CONCLUSIONS: The findings highlight the relevance of the neonatal individual characteristics of reactivity for identifying more vulnerable infants for future problems in biobehavioral regulation. PMID- 19560294 TI - Quantitative colour analysis of beach and dune sediments for forensic applications: a Portuguese example. AB - In this study, the colour of 23 sediment samples collected in different Portuguese beaches in the North (Entre-Douro and Minho) and 33 in the South (Algarve) was tested. Spectral colour parameters (L*a*b*) were measured after several presentation/pre-treatment methods: drying at 40 degrees C, sieving (<150 and <63 microm) and ashing at 850 degrees C, using a Konica Minolta CM-2600d spectrophotometer. A descriptive statistical analysis with L*a*b* values for each presentation/pre-treatment method was performed to ascertain within-sample reproducibility followed by a hierarchical cluster analysis to obtain significant discrimination between samples using colour analysis. The samples analysed presented a colour closest to red and yellow continuums and a lightness close to the pale side of the L*a*b* system colour sphere with samples collected in the North presenting analysis on average darker L values than the samples collected in the South. The cluster analysis, dried, unsieved bulk samples L*a*b* values gave the best discrimination between samples. The clusters analysis dried, unsieved bulk samples-dried, sieved <150 microm fraction set allowed better discrimination between samples compared with the dried, sieved <63 microm fraction set or dried, unsieved bulk samples-ashed bulk, unsieved samples set and even with dried, unsieved bulk samples/dried, sieved <150 microm fraction/dried, sieved <63 microm fraction/ashed bulk, unsieved samples considered all together. PMID- 19560295 TI - Medical negligence in drug associated deaths. AB - According to epidemiological studies adverse drug events are one of the most frequently encountered complications during medical treatment, a leading cause of hospitalisation and frequent cause of death. However, medical malpractice claims due to medication errors seem to be relatively rare. Based on a retrospective multicentre study on medical malpractice cases with lethal outcome (n=4450), drug related cases (n=575) were further evaluated. In 50% of cases a causal connection between drug therapy and death could be ruled out already after autopsy. In 232 cases a causal connection between drug therapy and death could be approved (drug allergies, relative overdose, wrong application, mix-up of drugs and sepsis after injection abscess). However, within the legal context only in 70 cases a medication error was approved which was in 42 cases causal for death, in 28 not. Administration of contraindicated drugs, incorrect application and relative overdose in renal insufficiency are the prevalent mistakes. Concerning the frequency of ADE in epidemiological studies medication errors are underreported in all data sources on medical malpractice; this seems to be due to the fact that even doctors and attending physicians rarely recognize an ADE; furthermore approving the connection between drug effect and death is extremely difficult for the expert witness. PMID- 19560296 TI - Mast cell reactivity at the margin of human skin wounds: an early cell marker of wound survival? AB - Detecting the vitality of mechanical skin wounds (antemortem versus postmortem injury) in human cadavers remains a specifically forensic problem. To determine whether skin mast cells (MCs) are activated during the very early phase of human wound healing we performed a histomorphometric evaluation of the extent of MC enzyme loss as an indication of MC degranulation at the wound margins of skin wounds in 64 human cadavers. We compared the number of tryptase-reactive MCs, which are said not to loose all of their enzyme activity during degranulation process, with the number of naphthol AS-D chloroacetate esterase (NAS-DClAE) positive MCs, which loose their complete enzyme activity in the form of enzyme positive granula after activation. The enzyme activity was evaluated on sequential histological sections after autopsy as an indirect quantification of the number of degranulated MCs. Most of the victims had died within 10-60 min after injury (n=50), 12 survived between 60 min and 24h, and only 2 victims survived more than 24h (12 days each). The number of enzyme-positive MCs were counted in six successive visual fields (0.785 mm(2)) on the one hand located parallel to and--on the other hand--at increasing distances outward from the wound margins. In victims surviving the injury less than 60 min the average number of NAS-DClAE-reactive MCs next to the wound margin was significantly lower than the number of tryptase-reactive MCs. The extent of the reduction in NAS DClAE-reactive MC counts correlated inversely with the distance from the wound edges. Our findings show that MCs undergo very early loss of NAS-DClAE activity at wound margins, and thus appear to be an early cell marker of wound survival. However, definitive evidence that the enzyme loss (degranulation) represents a vital process can only be obtained by comparing MC enzyme loss induced by injury during intact circulation with the MC reaction to injury inflicted very shortly after cardiac arrest, a question that can only be answered by animal experiments. PMID- 19560297 TI - About the origin of the sinus node artery. PMID- 19560298 TI - DPOAE in estimation of the function of the cochlea in tinnitus patients with normal hearing. AB - OBJECTIVE: The most probable place generating tinnitus in the auditory pathway is the outer hair cells (OHCs) inside the cochlea. Otoacoustic emissions are used to assess their activity. The objective of the investigation was to measure the features of distortion product otoacoustic emissions (DPOAE) in a group of tinnitus patients without hearing loss, estimate the diagnostic value of the parameters for the analysis of cochlear function in the patients, emphasizing those most useful in localizing tinnitus generators, and determine the hypothetical influence of hyperacusis and misophony on DPOAE parameters in tinnitus patients. PATIENTS AND METHODS: The material consisted of 44 patients with tinnitus and without hearing loss. In the control group were 33 patients without tinnitus with the same state of hearing. The tinnitus patients were divided into three subgroups: those with hyperacusis, those with misophonia, and those with neither. After collecting medical history and performing clinical examination of all the patients, tonal and impedance audiometry, ABR, and discomfort level were evaluated. Then DPOAE were measured using three procedures. First the amplitudes of two points per octave were assessed, second the "fine structure" method with 16-20 points per octave (f2/f1=1.22, L1=L2=70 dB), and the third procedure included recording the growth function in three series for input tones of f2=2002, 4004, and 6006Hz (f2/f1=1.22) and L1=L2 levels increasing by increments of 5 dB in each series. RESULTS AND CONCLUSIONS: Hyperacusis was found in 63% and misophonia in 10% of the tinnitus patients with no hearing loss. DPOAE amplitudes in recordings with two points per octave and the fine structure method are very valuable parameters for estimating cochlear function in tinnitus patients with normal hearing. Function growth rate cannot be the only parameter in measuring DPOAE in tinnitus patients, including subjects with hyperacusis and misophonia. The markedly higher DPOAE amplitudes in the group of tinnitus patients without hearing loss suggest that tinnitus may be caused by increased motility of the OHCs induced by decreasing efferent fiber activity, and not by OHC failure. Hyperacusis significantly increases the amplitude of DPOAE in tinnitus patients with no hearing loss. PMID- 19560299 TI - Results after supracricoid horizontal partial laryngectomy. AB - OBJECTIVES: This study analyzed oncological and functional results of supracricoid horizontal partial laryngectomy. METHODS: A retrospective study was conducted involving 20 patients with squamous cell carcinoma (SCC) of the larynx who underwent SCPL between 1996 and 2005 in Faculty of Medical Sciences of Santa Casa Hospital of Sao Paulo, Brazil. There were 18 male and 2 female patients with ages ranging from 39 to 74 years (median=58 years), of whom 19 were smokers and 14 alcoholics. The tumors were present in the glottis in 16 cases and supraglottis in 4; 5 were stage I or II and 15 were stage III or IV. We analyzed treatment given when rehabilitation was unsuccessful, oncological results of SCPL, including local and regional recurrences, time to recurrence and treatment given, distal metastases, global survival, survival free of disease, and appearance of second primary tumors. We also calculated the index of functional preservation of the larynx. RESULTS: Rehabilitation of swallowing capabilities and speech was achieved in 18 patients. Removal of the tracheostomy varied between 1 and 9 months. Rehabilitation was unsuccessful in two patients. Three patients required a total laryngectomy, two for unsuccessful rehabilitation and one for recurrence. The preservation of a functional larynx was 85%, with 10% of patients requiring a total laryngectomy after failed rehabilitation. CONCLUSIONS: Supracricoid horizontal partial laryngectomy is an efficient surgical oncology technique that yields good functional results for the treatment of laryngeal cancer. PMID- 19560300 TI - Adenocarcinoma of the paranasal sinuses and nasal cavity with lung metastasis showing complete response to combination chemotherapy with docetaxel, cisplatin and 5-fluorouracil (TPF): a case report. AB - Malignant tumors in the paranasal sinuses and nasal cavity are mostly squamous cell carcinomas, with comparatively few adenocarcinomas. Adenocarcinoma developing in paranasal sinuses and nasal cavity generally has a low response to radiotherapy and low chemotherapeutic sensitivity, making surgery the most reliable treatment. However, advanced adenocarcinoma is often difficult to treat due to anatomical complexity, and the outcome may not be satisfactory. We report the case of a 62-year-old man who presented with nasal congestion and epistaxis. We were unable to observe an oncogenic lesion in the nasal cavity directly, but the tumor gave inhomogeneous low intensity signals on T1-weighted images and an isointense signal on T2-weighted images in MRI. Adenocarcinoma was diagnosed by open biopsy. Six courses of chemotherapy by super-selective intra-arterial infusion of cisplatin with concurrent intensity-modulated radiation therapy of 70Gy were performed at another hospital. However, the tumor enlarged and developed distant metastasis to the lung after this therapy. Therefore, TPF chemotherapy (docetaxel, cisplatin and 5-fluorouracil) was performed 5 times, after which the tumor gradually reduced in size. The patient is alive after a follow-up period of 43 months, indicating that TPF chemotherapy may be useful for adenocarcinoma of the paranasal sinuses and nasal cavity. PMID- 19560301 TI - Characterization of new types of stationary phases for fast liquid chromatographic applications. AB - The performance of a narrow bore silica based monolith column (5 cm x 2 mm) was compared to 5 cm long narrow bore (internal diameter < or = 2.1 mm) columns, packed with shell particles (2.7 microm) and totally porous sub-2 microm particles (1.5 microm, 1.7 microm and 1.9 microm) in gradient and isocratic elution separations of steroids. The highest peak capacity could be achieved with the column packed with 1.5 microm totally porous particles. The columns packed with porous 1.7 microm and shell 2.7 microm particles showed very similar capacity. The monolith column provided the lowest capacity during gradient elution. The plate height (HETP) of the 2.7 microm Ascentis Express column was very similar to the HETP obtained with 1.5 microm and 1.7 microm totally porous particles. The Chromolith monolithic column displayed an efficiency that is comparable to that of columns packed with spherical particles having their diameter between 3 microm and 4 microm. A kinetic plot analysis is presented to compare the theoretical analysis speed of different separation media. At 200 bar, the monolith column provided the highest performance when the required plate number was higher than 5000 (N>5000), however the efficiency drifted off faster in the range of N<5000 than in the case of packed columns. If the possibility of maximum performance was utilized (1000 bar for sub-2 microm particles, 600 bar for shell particles and 200 bar for monolith column) the monolith column would provide the poorest efficiency, while the column, packed with 1.5 microm particles offered the shortest impedance time. PMID- 19560302 TI - Quinoline alkaloids in honey: further analytical (HPLC-DAD-ESI-MS, multidimensional diffusion-ordered NMR spectroscopy), theoretical and chemometric studies. AB - The wound-healing properties of honey are well established and it has been suggested that, among its pharmaco-active constituents, kynurenic acid (KA) exerts antinociceptive action on injured tissue by antagonizing NMDA at peripheral GABA receptors. The aim of this study was to investigate the quantitative profile of KA and of two recently identified, structurally related derivatives, 3-pyrrolidinyl-kynurenic acid (3-PKA) and its gamma-lactamic derivative (gamma-LACT-3-PKA), by examining their mass spectrometric behavior, in honeys from different botanical sources. We used a combination of HPLC-DAD-ESI-MS and NMR techniques (one-dimensional (1)H NMR and diffusion-ordered spectroscopy NMR). Chestnut honey constantly contained KA (2114.9-23 g/kg), 3-PKA (482.8-80 mg/kg) and gamma-LACT-3-PKA (845.8-32 mg/kg), confirming their reliability as markers of origin. A new metabolite, 4-quinolone (4-QUIN), was identified for the first time in one chestnut honey sample (743.4 mg/kg). Small amounts of KA were found in honeydew, sunflower, multifloral, almond and eucalyptus honeys, in the range of 23.1-143 mg/kg, suggesting contamination with chestnut honey. Total phenol content (TPC) was in the range from 194.9 to 1636.3 mg(GAE)/kg and total antiradical activity (TAA) from 61 to 940 mg/(GAE)/kg), depending on the botanical origin. Principal component analysis (PCA) was then done on these data. The three different clusters depicted: (i) antinociceptive activity from KA and/or its derivatives, typical of chestnut honey; (ii) antioxidant/radical scavenging activity by antioxidants responsible for the antiinflammatory action (dark honeys); (iii) peroxide-dependent antibacterial activity due to H(2)O(2) production by glucose oxidase in honey. The PCA findings provide useful indications for the dermatologist for the treatment of topical diseases, and the profiling of KA and its derivatives may shed light on new aspects of the kynurenine pathway involved in tryptophan metabolism. PMID- 19560303 TI - Validation of a preparation for decision making scale. AB - OBJECTIVE: The Preparation for Decision Making (PrepDM) scale was developed to evaluate decision processes relating to the preparation of patients for decision making and dialoguing with their practitioners. The objective of this study was to evaluate the scale's psychometric properties. METHODS: From July 2005 to March 2006, after viewing a decision aid prescribed during routine clinical care, patients completed a questionnaire including: demographic information, treatment intention, decisional conflict, decision aid acceptability, and the PrepDM scale. RESULTS: Four hundred orthopaedic patients completed the questionnaire. The PrepDM scale showed significant correlation with the informed (r=-0.21, p<0.01) and support (r=-0.13, p=0.01) subscales (DCS); and discriminated significantly between patients who did and did not find the decision aid helpful (p<0.0001). Alpha coefficients for internal consistency ranged from 0.92 to 0.96. The scale is strongly unidimensional (principal components analysis) and Item Response Theory analyses demonstrated that all ten scale items function very well. CONCLUSION: The psychometric properties of the PrepDM scale are very good. PRACTICE IMPLICATIONS: The scale could allow more comprehensive evaluation of interventions designed to prepare patients for shared-decision making encounters regarding complex health care decisions. PMID- 19560304 TI - From patient talk to physician notes-Comparing the content of medical interviews with medical records in a sample of outpatients in Internal Medicine. AB - OBJECTIVES: An increasing number of consultations are delivered in group practices, where a stable 1:1 relationship between patient and physician cannot be guaranteed. Therefore, correct documentation of the content of a consultation is crucial to hand over information from one health care professional to the next. METHODS: We randomly selected 20 interviews from a series of 56 videotaped consultations with patients requesting a general check-up exam in the outpatient department of Internal Medicine at the University Hospital Basel. All patients actively denied having any symptoms or specific health concerns at the time they made their appointment. Videotapes were analysed with the Roter Interaction Analysis System (RIAS). Corresponding physician notes were analysed with a category check-list that contained the information related items from RIAS. RESULTS: Interviews contained a total of 9.002 utterances and lasted between 15 and 53min (mean duration: 37min). Patient-centred communication (Waiting, Echoing, Mirroring, Summarising) in the videos significantly correlated with the amount of information presented by patients: medical information (r=.57; p=.009), therapeutic information (r=.50; p=.03), psychosocial information (r=.41; p=.07), life style information (r=.52; p=.02), and with the sum of patient information (r=.64; p=.003). Even though there was a significant correlation between the amount of information from the video and information in physician's notes in some categories (patient gives medical information; Pearson's r=.45; p=.05, patient gives psychosocial information; Pearson's r=.49; p=.03), an inspection of the regression lines shows that a large extent of patient information is omitted from the charts. Physicians never discussed with patients whether information should be documented in the charts or omitted. CONCLUSIONS: The use of typical patient centred techniques increases information gathered from patients. Physicians document only a small percentage of patient information in the charts, their 'condensing heuristic' is not shared with patients. PRACTICE IMPLICATIONS: Patient involvement should be advocated not only to medical decision making but also to the way physicians document the content of a consultation. It is a joint responsibility of patient and health care professional to decide, which information should be kept and thus be communicated to another health care professional in future consultations. PMID- 19560305 TI - Will the introduction of non-invasive prenatal diagnostic testing erode informed choices? An experimental study of health care professionals. AB - OBJECTIVE: Informed choice is a fundamental concept within prenatal care. The present study assessed the extent to which the introduction of non-invasive prenatal diagnosis (NIPD) of Down's syndrome may undermine the process of making informed choices to undergo prenatal testing or screening for Down's syndrome by altering the quality and quantity of pre-test counselling. METHODS: 231 obstetricians and midwives were randomly allocated one of three vignettes, each describing a different type of test: (a) invasive prenatal diagnosis (IPD), (b) non-invasive prenatal diagnosis (NIPD) or (c) Down's syndrome screening (DSS). Participants were then asked to complete a questionnaire assessing (1) the information considered important to communicate to women, (2) whether test offer and uptake should take place on different days, and (3) whether signed consent forms should be obtained prior to testing. RESULTS: Across the three test types, five out of the seven presented topics were considered equally important to communicate, including the information that testing is the woman's choice. Compared with participants receiving the IPD vignette, those receiving the NIPD and DSS vignettes were less likely to report that counselling and testing should occur on different days (IPD 94.7% versus 74.1% and 73.9% for NIPD and DSS respectively, p=.001) and that written consent was a necessity (IPD 96.1% versus 68.3% and 75.4% for NIPD and DSS respectively, p<.001). CONCLUSION: This study provides the first empirical evidence to demonstrate that practitioners may view the consent process for NIPD differently to IPD. There is potential for the introduction of NIPD to undermine women making informed choices in the context of prenatal diagnostic testing for conditions like DS. PRACTICE IMPLICATIONS: Given the importance of informed choice in reproductive decision-making, implementation of any programme based on NIPD should be designed to facilitate this. PMID- 19560306 TI - Parents' contrasting views on diet versus activity of children: implications for health promotion and obesity prevention. AB - OBJECTIVE: To investigate parents' perceptions of, and contributions to, food and activity choices and maintenance of a healthy lifestyle in children. METHODS: Ethnographically informed qualitative study using in-depth semi-structured interviews with 47 parents (32 mothers and 15 fathers) of 5-7-year-old children. RESULTS: Parents were concerned about children's low intake of food, the development of eating disorders and children being 'too active'. Therefore, they promoted eating and were controlling of diet because they thought that children would not eat enough for healthy development. They did not promote, or even curbed, physical activity because they considered their children were already active. Their accounts suggested lack of awareness of the dangers of over consumption and inactivity. CONCLUSIONS: Parents' perceptions affected the way they raised their children and the choices that they made for them. Parents' concerns about under-consumption of food and over-activity contrast with the public health priorities to reduce intake and promote exercise. PRACTICE IMPLICATIONS: Health professionals need to take into account parents' perspectives on diet and physical activity when attempting to promote children's health and prevent obesity. PMID- 19560308 TI - Intravenous leiomyomatosis with inferior vena cava and heart extension. AB - BACKGROUND: Intravenous leiomyomatosis (IVL) is a rare tumor that is histologically benign but biologically malignant. Less than 200 cases of IVL have been reported, most of them were individual reports. METHODS: Six patients with IVL involving the inferior vena cava were analyzed. RESULTS: Three patients received one-stage operations and two received two-stage operations. All operations were successful. No perioperative death or other complications were observed. Primary tumors and intravenous tumorous emboli were completely resected from four patients. Residual tumor remained in one patient who had serious adhesions due to multiple previous surgeries; however, with antiestrogen therapy, the residual tumor significantly regressed. All patients had tumor relapse after the operation. CONCLUSIONS: IVL should be given more attention by vascular surgeons, although it is extremely rare. Many therapeutic methods are available for uterine leiomyomatosis involving inferior vena cava, among which operation is the best choice. PMID- 19560307 TI - Lessons learned from an HIV adherence pilot study in the Deep South. AB - OBJECTIVE: Adherence to treatment for chronic illnesses, including HIV disease, is a complex process, and needs practical interventions in poorly resourced clinic settings. METHODS: This study tested the feasibility of an adherence intervention in 73 HIV-infected individuals in a Deep South public clinic based on Fisher & Fisher's Information-Motivation-Behavioral Skills Model. RESULTS: There was high baseline adherence and unexpectedly high clinic attrition, and 27% of the intervention group received less than one-quarter of the planned intervention contacts. Refill rate was the adherence measure that correlated best with HIV viral load and CD4 count, and there was poor use of electronic adherence monitoring (MEMS). Interviewed individuals expressed positive feelings about audio-supported computer-assisted survey instruments (ACASI) and the intervention support. CONCLUSIONS: This process evaluation showed feasible study components in this population and setting. Lessons learned included: (1) clinic retention is an important part of adherence; (2) telephone interventions may need to add additional technology and flexibility to maximize dose; (3) ongoing fidelity monitoring is important with motivational interviewing; (4) refill rate was the most accurate adherence assessment; (5) MEMS was not well-accepted; (6) ACASI was easily used in this population; and (7) individuals appreciated adherence support from a consistent caring individual. PMID- 19560309 TI - Persistent sciatic artery aneurysm treated with concomitant tibial bypass and vascular plug embolization. AB - The presence of a persistent sciatic artery is a rare congenital vascular malformation. Complications associated with aneurysmal degeneration of this aberrant vessel include rupture, thrombosis, and embolization with obliteration of outflow vessels. We describe the case of an 82-year-old female presenting with critical limb ischemia due to embolization from a partially thrombosed persistent sciatic artery aneurysm. Successful treatment was achieved via a common femoral to posterior tibial artery bypass with the great saphenous vein and vascular plug embolization of the aneurysm. PMID- 19560310 TI - Perioperative asymptomatic cardiac damage after endovascular abdominal aneurysm repair is associated with poor long-term outcome. AB - BACKGROUND: Endovascular abdominal aortic aneurysm (AAA) repair (EVAR) is associated with a decreased incidence of perioperative cardiac complications compared with open repair. However, EVAR is not associated with long-term survival benefit. This study assessed the effect of perioperative asymptomatic cardiac damage after EVAR on long-term prognosis. METHODS: In 220 patients undergoing elective EVAR, routine sampling for levels of cardiac troponin T and electrocardiography (ECG) were performed on days 1, 3, and 7 during the patient's hospital stay. Elevated cardiac troponin T was defined as serum concentrations >or=0.01 ng/mL. Asymptomatic cardiac damage was defined as cardiac troponin T release without symptoms or ECG changes. The median follow-up was 2.9 years. Survival status was obtained by contacting the Office of Civil Registry. RESULTS: Release of cardiac troponin T (median, 0.08 ng/mL) occurred in 24 of 220 patients, of whom 20 (83%) were asymptomatic and without ECG changes. Patients with asymptomatic cardiac damage had a mortality rate of 49% [corrected] after 2.9 years vs 15% [corrected] for patients without perioperative cardiac damage (P < .001). Also after adjustment for clinical risk factors and medication use applying multivariate Cox regression analysis, asymptomatic cardiac damage was associated with a 2.3-fold increased risk for death (95% confidence interval, 1.1 5.1). Statin use was associated with a reduced long-term risk for death (hazard ratio, 0.5; 95% confidence interval, 0.3-0.9). CONCLUSION: Asymptomatic cardiac damage in patients undergoing EVAR is associated with poor long-term outcome. Routine perioperative cardiac screening after EVAR might be warranted. PMID- 19560311 TI - Long-term results of balloon-expandable LifePath endografts in abdominal aortic aneurysm: a single-center experience. AB - OBJECTIVE: This retrospective single-center study analyzed long-term results after LifePath (Edwards Lifesciences LLC, Irvine, Calif) endoprosthesis implantation for abdominal aortic aneurysm (AAA), primarily focusing on the wire form fracture issue and consecutive endoleak rate. METHODS: Between 1999 and 2004, all consecutive patients with LifePath AAA devices in our institution were included in the retrospective analysis. All patients had computed tomography angiography (CTA) imaging preoperatively and image postprocessing. The follow-up using CTA imaging specifically addressed material fatigue (wire form fractures) resulting in migrations and type I endoleaks. RESULTS: During the 6-year study period, which included the 1-year withdrawal and redesign of the device, 51 patients were treated with LifePath AAA endografts. The 30-day mortality was 0%. The perioperative 30-day morbidity was 9.8%. One patient required a primary conversion due to misdeployment of the iliac limbs within the graft main body. The primary endoleak rate was 20.56% (type I, 2%; type II, 19.6%). During the mean follow-up of 40.7 months, 12 patients died, six were lost to follow-up, and 32 underwent subsequent CTA imaging. Eight patients (25%) demonstrated a proximal type I endoleak, seven (22%) had a type II endoleak, and three had a type III endoleak (9%). In nine patients (28.1%), wire form fractures could be detected at image postprocessing. Four patients required a secondary conversion due to endoleak and aneurysm growth (2 type I endoleaks and 2 type III endoleaks). CONCLUSION: Wire form fracture is the major structural problem in the LifePath balloon-expandable endograft device, resulting in a significant endoleak rate. We must caution those patients with a LifePath device in-situ that careful follow-up must be performed due to material fatigue and they should consider secondary conversion. PMID- 19560312 TI - Screening of unrecognized peripheral arterial disease (PAD) using ankle-brachial index in high cardiovascular risk patients free from symptomatic PAD. AB - OBJECTIVE: To determine the utility of ankle-brachial index (ABI) in screening for unrecognized peripheral arterial disease (PAD). Although PAD is a consistent predictor of cardiovascular morbidity and mortality, it is often under-diagnosed and under-treated. METHODS: In this prospective, observational, real-life, epidemiologic study (ELLIPSE) the prevalence of PAD (ABI < 0.9) was calculated in 2146 asymptomatic patients > or =55 years of age who were at high cardiovascular risk and who were hospitalized in departments of cardiology, diabetology, geriatrics, internal medicine, or neurology in metropolitan France. Univariate and multivariate analyses were performed to identify PAD risk factors. The discriminatory power of the model was evaluated by calculating the area under the curve (AUC) of the receiver operating characteristic curve. RESULTS: The ABI was <0.9 in 41.1% of patients. In the multivariate analysis, absence of > or =1 pulse (odds ratio [OR], 2.18; 95% confidence interval [CI], 1.81 to 2.63; P < .0001), arterial bruit (OR, 1.92; 95%CI, 1.34 to 2.75; P < .0004), previous non-Q-wave myocardial infarction (OR, 1.50; 95%CI, 1.08 to 2.08; P = .02), regular smoking (OR, 1.49; 95%CI, 1.22 to 1.80; P < .0001), age > or =81 years (OR, 1.45; 95%CI, 1.15 to 1.82; P = .001), creatinine clearance <60 mL/min (OR, 1.33; 95%CI, 1.08 to 1.63; P = .008), and treated hypertension (OR, 1.28; 95%CI, 1.03 to 1.59; P = .03) were significantly associated with PAD. Although risk increased with the number of variables, the model, based on clinical symptoms and on medical history parameters, was not discriminatory (AUC = 0.66). On average, physicians took 15 minutes to perform the ABI test. CONCLUSIONS: The high prevalence of asymptomatic PAD in this patient population suggests that ABI should systematically be performed in high-risk hospitalized patients to ensure that appropriate secondary prevention programs are initiated. PMID- 19560313 TI - Age-related trends in utilization and outcome of open and endovascular repair for abdominal aortic aneurysm in the United States, 2001-2006. AB - OBJECTIVE: This study used a large national administrative in-hospital database to compare utilization and age-specific outcomes between open repair (OAR) and endovascular (EVAR) repair for the treatment of abdominal aortic aneurysm (AAA). METHODS: Discharges with the principal International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) procedure codes for EVAR and OAR and principal diagnosis code of intact AAAs were selected from the 2001 to 2006 Nationwide Inpatient Sample (NIS). Weighted least-square regression was used to test the trend of utilization by age. Multiple linear and logistic regression analyses were used to assess the risk-adjusted outcomes. RESULTS: Nationally, the estimated number of elective AAAs treated with EVAR increased from 11,171 in 2001 to 21,725 in 2006 (P = .003). The number of elective AAAs treated with OAR declined from 17,784 to 8451 during the same period (P < .001). By 2006, EVAR was more frequently used than OAR for patients of all ages. Compared with the younger age groups, patients aged >or=85 years had a significant increase in the total number of asymptomatic AAA repairs, driven almost entirely by an increase in the use of EVAR. Compared with open patients, EVAR patients had a significantly shorter length of hospitalization (adjusted mean, 2.99 days [95% confidence interval (CI), 2.97-3.01] vs 8.78 days [95% CI, 8.53-8.57]), less in-hospital mortality (odds ratio [OR], 0.23; 95% CI, 0.19-0.28), fewer in-hospital complications (OR, 0.27; 95% CI, 0.25-0.28), and a higher likelihood of being discharged to home (OR, 3.95; 95% CI, 3.62-4.31). The reduction of complications from the use of EVAR versus OAR was most dramatic for the oldest patients. CONCLUSIONS: As short-term surgical outcomes are consistently improving for patients undergoing AAA repair, elective EVAR has replaced OAR as the more common method of repair in the United States. The introduction of this technology has been rapidly adopted, particularly for the oldest-old surgical patients, aged >or=85 years, who previously may not have been offered surgical intervention for asymptomatic AAA. Further investigation is necessary to examine whether this trend improves the long-term survival and quality of life for this elderly population. PMID- 19560314 TI - Measuring health anxiety: moving past the dichotomous response option of the original Whiteley Index. AB - The Whiteley Index [WI; Pilowsky, I. (1967). Dimensions of hypochondriasis. British Journal of Psychiatry, 113, 89-93] is a widely used self-report measure of health anxiety, hypochondriasis, and illness phobia; however, the factor structure of the WI has proven unstable. Moreover, factorial investigations of the WI persist in the original true/false response format despite criticisms of dichotomous scales and the availability of a Likert scale version [Barsky, A. J., Cleary, P. D., Wyshak, G., Spitzer, R. L., Williams, J. B. W., & Klerman, G. L. (1992). A structured diagnostic interview for hypochondriasis: a proposed criterion standard. The Journal of Nervous and Mental Disease, 180, 20-27]. The current investigation explores the factor structure of the 5-point Likert scale version of the WI using a sample of 287 students (64 men, ages 18-34 [M=20.6; SD=3.2]; 223 women ages 18-45 [M=20.2; SD=3.2]) divided into two sex-matched groups. A 2-factor 8-item model was extracted from the first group using exploratory factor analysis. However, confirmatory factor analysis with the second group showed superior fit to the data using a precedent 2-factor 6-item model, demonstrating factorial invariance across response options. Comprehensive results, implications, and directions for future research are discussed. PMID- 19560315 TI - A multi-ethnic examination of socioevaluative fears. AB - Established and emerging cognitive models of social anxiety have provided researchers and clinicians with a solid foundation for understanding and treating this phenomenon. Much of the support for these models, however, has been derived from predominantly Caucasian samples. While the evidence supports the concept of socioevaluative fears as being universal, ethnic/cultural influences can dramatically alter the cognitive profile of the fears. The purpose of this study was to examine the cross-ethnic equivalence of the bivalent fear of evaluation model of social anxiety among an ethnically diverse sample of 799 undergraduate students from the United States. A series of confirmatory factor analyses indicated good model fit for each of the ethnic groups examined, and that holding factor loadings and latent variances and covariances equivalent did not alter model fit significantly. PMID- 19560316 TI - The concurrent validity of a hand-held versus a stationary dynamometer in testing isometric shoulder strength. AB - STUDY DESIGN: Clinical Measurement-Validity. INTRODUCTION: Validity of the JTech PowerTrack II hand-held dynamometer (JTech; JTech Medical, Salt Lake City, UT) for measuring shoulder strength has yet to be established. PURPOSE OF THE STUDY: To examine the concurrent validity of isometric strength scores obtained with the JTech PowerTrack II, and on a stationary dynamometer, the LIDO WorkSET (LIDO; LoredanBiomedical, West Sacramento, CA). METHODS: Thirty-eight subjects performed three maximal efforts of shoulder flexion, abduction, and external rotation on a single occasion on the two dynamometers. Two testers were randomly assigned to administer the tests. RESULTS: Pearson correlations between the scores on the two dynamometers (r.0.81) indicated a good concurrent validity. Correlations were similar when the results were subdivided by tester or gender. CONCLUSIONS: This study suggests that either the JTech PowerTrack II or LIDOWorkSET provide comparable scores for shoulder strength. Although not interchangeable because of the differences in units of measurement, the relative conclusions about strength should be similar, regardless of which instrument is used. LEVEL OF EVIDENCE: Not applicable. PMID- 19560318 TI - Upper limb neurodynamic test of the radial nerve: a study of responses in symptomatic and asymptomatic subjects. AB - STUDY DESIGN: Clinical measurement. INTRODUCTION: Nonspecific cervical pain is a common clinical presentation. The role of upper limb neurodynamic tests (ULNT), for evaluation and treatment intervention, is not well defined for this population. PURPOSE OF THE STUDY: This study's purpose was to determine if the radial-biased (RB)-ULNT discriminates any response differences between symptomatic subjects with a positive (+) RB-ULNT (n=36), symptomatic subjects with a negative (-) RB-ULNT (n=24), and asymptomatic subjects (n=60). METHODS: Sixty asymptomatic and 60 subjects presenting with nonspecific cervical and/or unilateral upper extremity pain were compared using the RB-ULNT. Symptomatic subjects were further divided in (+) and (-) RB-ULNT groups due to their response to the RB-ULNT. Within the symptomatic population, a positive response to the RB ULNT was defined by the symptomatic subject reporting their sensations were increased with contralateral cervical lateral flexion and decreased with ipsilateral cervical lateral flexion. Sensation provocation and location were evaluated using the RB-ULNT in all the subjects during each stage of the testing. RESULTS: Significant differences on stage of reproduction and type of sensations were identified between 1) the (+) RB-ULNT symptomatic subjects, 2) the (-) RB ULNT symptomatic subjects, and 3) the asymptomatic subjects. The (+) RB-ULNT group showed significantly increased pain responses during the first stage of the RB-ULNT compared with the (-) RB-ULNT group and the asymptomatic subjects. The (+) RB-ULNT also showed significantly decreased glenohumeral abduction passive range of motion when compared with the asymptomatic group. CONCLUSION: Clinically, the differences found between the groups in their response to the RB ULNT suggest heightened mechanosensitivity in the (+) RB-ULNT group. LEVEL OF EVIDENCE: 3a. PMID- 19560317 TI - Relationship between patient satisfaction and objective functional outcome after surgical treatment for distal radius fractures. AB - STUDY DESIGN: Prospective cohort. INTRODUCTION: Patient satisfaction is increasingly used as a metric of health care outcomes. The relationship between patient satisfaction and functional outcomes metrics is understudied. PURPOSE: To determine the minimum recovery needed in grip strength, key pinch strength, and arc of motion needed for patient satisfaction after treatment of distal radius fracture (DRF) with volar locking plating system placement. METHODS: A prospective cohort of 125 DRF patients was evaluated three months after surgery for grip strength, key pinch strength, wrist arc of motion, and satisfaction with hand strength and wrist arc of motion. Receiver operating characteristic curves were constructed using patient satisfaction items as the "gold standard" and each functional measure of outcome as a predictor. RESULTS: We found that the optimal cutpoints to distinguish satisfaction from dissatisfaction occurred when patients had recovered 65% of their grip strength, 87% of their key grip strength, and 95% of the wrist arc of motion, as measured as percents of their uninjured wrists. CONCLUSIONS: A much greater wrist range of motion must be recovered for patients to be satisfied than what is needed to perform activities of daily living. LEVEL OF EVIDENCE: Diagnosis level 2. PMID- 19560319 TI - Focal Cortical Dysplasia (FCD) lesion analysis with complex diffusion approach. AB - Identification of Focal Cortical Dysplasia (FCD) can be difficult due to the subtle MRI changes. Though sequences like FLAIR (fluid attenuated inversion recovery) can detect a large majority of these lesions, there are smaller lesions without signal changes that can easily go unnoticed by the naked eye. The aim of this study is to improve the visibility of focal cortical dysplasia lesions in the T1 weighted brain MRI images. In the proposed method, we used a complex diffusion based approach for calculating the FCD affected areas. Based on the diffused image and thickness map, a complex map is created. From this complex map; FCD areas can be easily identified. MRI brains of 48 subjects selected by neuroradiologists were given to computer scientists who developed the complex map for identifying the cortical dysplasia. The scientists were blinded to the MRI interpretation result of the neuroradiologist. The FCD could be identified in all the patients in whom surgery was done, however three patients had false positive lesions. More lesions were identified in patients in whom surgery was not performed and lesions were seen in few of the controls. These were considered as false positive. This computer aided detection technique using complex diffusion approach can help detect focal cortical dysplasia in patients with epilepsy. PMID- 19560320 TI - SPM analysis of ictal-interictal SPECT in mesial temporal lobe epilepsy: relationships between ictal semiology and perfusion changes. AB - A combination of temporo-limbic hyperperfusion and extratemporal hypoperfusion was observed during complex partial seizures (CPS) in temporal lobe epilepsy (TLE). To investigate the clinical correlate of perfusion changes in TLE, we analyzed focal seizures of increasing severity using voxel-based analysis of ictal SPECT. We selected 26 pre-operative pairs of ictal-interictal SPECTs from adult mesial TLE patients, seizure-free after surgery. Ictal SPECTs were classified in three groups: motionless seizures (group ML, n=8), seizures with motor automatisms (MA) without dystonic posturing (DP) (group MA, n=8), and seizures with DP with or without MA (DP, n=10). Patients of group ML had simple partial seizures (SPS), while others had CPS. Groups of ictal-interictal SPECT were compared to a control group using statistical parametric mapping (SPM). In ML group, SPM analysis failed to show significant changes. Hyperperfusion involved the anteromesial temporal region in MA group, and also the insula, posterior putamen and thalamus in DP group. Hypoperfusion was restricted to the posterior cingulate and prefrontal regions in MA group, and involved more widespread associative anterior and posterior regions in DP group. Temporal lobe seizures with DP show the most complex pattern of combined hyper-hypoperfusion, possibly related both to a larger spread and the recruitment of more powerful inhibitory processes. PMID- 19560321 TI - Relationship between psychotic and obsessive compulsive symptoms in schizophrenia. AB - The presence of obsessive compulsive symptoms (OCSs) in schizophrenia was recognized as early as the first descriptions of the illness. Studies investigating the association between OCSs and schizophrenia have defined their co-occurrence in terms of co-morbidity and compared schizophrenia patients separated into groups according to whether they presented OCSs or not. However, most of these studies did not take both the complexity of the schizophrenia phenomenology and that of OCSs into account. The present research investigates the relationship between schizophrenia symptoms and OCSs using a correlational approach with a dimensional perspective in order to determine how the OCSs contribute to symptom expression in schizophrenia. Fifty nine schizophrenia patients were rated for schizophrenia symptoms (SAPS-SANS) and OCSs (Y-BOCS). Schizophrenia symptoms scores were collapsed into four dimensional scores and OCSs into for other dimensional scores. The latter were entered as explanatory variables to determine their associations with schizophrenia dimension scores using series of stepwise regression models. The results showed a strong positive relationship between Delusions and Obsessions consistent with the view that they reflect manifestations of the similar mechanisms. Similar results indicate an association between Auditory hallucinations and Compulsions also suggesting that they share common mechanisms. On the other hand, there were inverse relationships between Somatic Obsessions and Disorganization and between Hoarding/Collecting Compulsions and Delusions or Auditory hallucinations. These results may reflect that these OCSs have a protective effect against disorganization and psychotic symptoms respectively. PMID- 19560322 TI - Paliperidone extended-release for schizophrenia: effects on symptoms and functioning in acutely ill patients with negative symptoms. AB - BACKGROUND: Most patients with schizophrenia exhibit negative symptoms, even during acute episodes. These difficult-to-treat symptoms are often associated with poor functioning and outcomes. METHODS: A post-hoc analysis of pooled data from three 6-week double-blind studies included patients in an acute episode of schizophrenia who received paliperidone extended-release (ER) (3, 6, 9, or 12 mg) or placebo. Based on criteria developed by the authors, patients were stratified by the presence or absence of predominant negative symptoms at baseline (>or=40% of the maximum negative factor score and <40% of the maximum positive factor score on the Positive and Negative Syndrome Scale [PANSS]). RESULTS: Although these studies were not designed to examine patients with predominant negative symptoms, the criteria identified 23% of acutely ill patients (270/1193). The mean (SD) baseline PANSS negative symptoms factor score, 27.4 (3.3), was 49% of the maximum; the positive symptoms factor score, 23.7 (2.8), was 33% of the maximum. Completion rates with paliperidone ER (n=195) and placebo (n=75) were 64.6% and 44.0%, respectively. Greater improvements occurred with paliperidone ER vs placebo on PANSS (total, negative and other factors), Clinical Global Impressions-Severity and Personal and Social Performance scores at endpoint (all P values <0.05). Adverse events reported in >or=10% of patients were (paliperidone ER vs placebo): headache (14.4% vs 6.7%), insomnia (13.8% vs 21.3%) and sinus tachycardia (10.3% vs 1.3%). Paliperidone ER treatment was associated with a similar response profile in patients without predominant negative symptoms (paliperidone ER, n=647; placebo, n=276). CONCLUSIONS: Schizophrenia patients with predominant negative symptoms were identified in a population of acutely ill patients. Findings of this post-hoc analysis suggest that acutely ill patients with or without predominant negative symptoms respond similarly to treatment with paliperidone ER. No unexpected tolerability findings were observed. PMID- 19560323 TI - Apolipoprotein E allele 4 is not a sufficient or a necessary predictor of the development of Mild Cognitive Impairment. AB - The presence of Mild Cognitive Impairment (MCI) and of an apolipoprotein E (apoE) varepsilon4 allele both predict the development of Alzheimer's disease. However, the extent to which this allele also predicts the development of MCI is unclear even though MCI is an early transitional stage in the development of Alzheimer's disease. The present study investigates the prevalence of the apoE varepsilon4 allele in incipient MCI. Participants were recruited from the population-based Leipzig Longitudinal Study of the Aged (LEILA75+). All subjects who were initially cognitively healthy, i.e. did not meet MCI criteria described by Petersen [Petersen RC. Mild cognitive impairment. J Intern Med 2004; 256(3): 183 94], and whose apoE status could be determined were followed-up. After 4.5 years, 15.5% of the cognitively healthy target population had developed MCI. The frequencies of the apoE varepsilon4 genotype did not differ between individuals with incipient MCI (12.9%) and individuals who remained cognitively healthy during the study (18.4%, p>0.5). Consequently, the apoE varepsilon4 genotype is not a necessary or sufficient risk factor for MCI. Further studies need to investigate the influence of the whole range of genetic and environmental risk factors on the course of Alzheimer's disease including the initial development of MCI and the later conversion to Alzheimer's disease. PMID- 19560324 TI - Calcein permeation across phosphatidylcholine bilayer membrane: effects of membrane fluidity, liposome size, and immobilization. AB - The permeation of calcein across the phospholipid bilayer membrane is a key phenomenon in the detection system using liposomes as a sensor unit. The behavior of the calcein release from the liposome was analyzed by a first-order kinetic to obtain the permeability coefficient, Ps [cm/s]. The Ps value for the neutral liposome, prepared by zwitterionic 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC), was found to depend on both the diameter of liposome and the temperature. The membrane fluidity of the POPC liposome, evaluated by the hydrophobic probe, 1-(4-trimethyl-aminophenyl)-6-diphenyl-1,3,5-hexatriene, was also dependent on the liposome diameter and the temperature. The Ps values for various neutral liposomes under gel phase or liquid-crystalline phase were correlated with their membrane fluidity, although some data were a little scattered, possibly due to the lamellarity. It is therefore considered that the membrane fluidity dominates the permeability of calcein across the neutral phospholipid membrane. Based on the above results, the Ps value for liposomes immobilized on the solid surface is discussed. PMID- 19560325 TI - Microanatomical and immunohistochemical study of the human radial nerve at the antecubital fossa. AB - BACKGROUND: Poor prognosis of radial nerve repair in elderly patients may be due to changes in intraneural anatomy with age. Also, chances of Complex Regional Pain Syndrome-Type I (CRPS-I) following radial nerve injury are comparatively high. The present study is to find the fascicular pattern of radial nerve (at antecubital fossa), microanatomic morphometric characteristics of its connective tissue components and changes with age and study of intraneural sympathetic fiber content. METHODS: Twenty human (21-87 years) cadaveric radial nerves have been collected from antecubital fossa and the study has been performed at magnifications (10x, 20x and 40x objective) after routine histological (hematoxylin & eosin stain) processing was done for morphometric analysis (total cross-sectional, fascicular and non-fascicular area) and immunohistochemical (tyrosine hydroxylase) processing for sympathetic fibers. RESULTS: The radial nerve is of a polyfascicular type with a grouped pattern of nerve fascicular distribution. The number of fascicles range from 9 to 17, whereas the number of fascicles per square millimeter of a cross-sectional area is 1.95. In elderly cases, there is significant increase in total radial nerve cross-sectional area due to an increase in its non-fascicular connective tissue area and excessive adipose tissue deposition in interfascicular domains. The average sympathetic fiber area is 0.046 mm(2) without definite relationship to age. CONCLUSION: There is an increase in total nerve cross-sectional area of the radial nerve in elderly cases. There is no relationship of sympathetic content to age. Our study makes an attempt to build a normal data base for radial nerve which might be helpful during the application of diagnostic procedures. PMID- 19560326 TI - Embryonic rotational behaviour in the pond snail Lymnaea stagnalis: influences of environmental oxygen and development stage. AB - Responses of freshwater organisms to environmental oxygen tensions (PO(2)) have focused on adult (i.e. late developmental) stages, yet responses of embryonic stages to changes in environmental PO(2) must also have implications for organismal biology. Here we assess how the rotational behaviour of the freshwater snail Lymnaea stagnalis changes during development in response to conditions of hypoxia and hyperoxia. As rotation rate is linked to gas mixing in the fluid surrounding the embryo, we predicted that it would increase under hypoxic conditions but decrease under hyperoxia. Contrary to predictions, early, veliger stage embryos showed no change in their rotation rate under hyperoxia, and later, hippo stage embryos showed only a marginally significant increase in rotation under these conditions. Predictions for hypoxia were broadly supported, however, with both veliger and hippo stages showing a marked hypoxia-related increase in their rotation rates. There were also subtle differences between developmental stages, with hippos responding at PO(2)s (50% air saturation) greater than those required to elicit a similar response in veligers (20% air saturation). Differences between developmental stages also occurred on return to normoxic conditions following hypoxia: rotation in veligers returned to pre-exposure levels, whereas there was a virtual cessation in embryos at the hippo stage, likely the result of overstimulation of oxygen sensors driving ciliary movement in later, more developed embryos. Together, these findings suggest that the spinning activity of L. stagnalis embryos varies depending on environmental PO(2)s and developmental stage, increasing during hypoxia to mix capsular contents and maintain a diffusive gradient for oxygen entry into the capsule from the external environment ("stir-bar" theory of embryonic rotational behaviour). PMID- 19560327 TI - D-camphor-crataegus berry extract combination increases blood pressure and cognitive functioning in the elderly - a randomized, placebo controlled double blind study. AB - The present study investigated whether the D-camphor-crataegus berry extract combination Korodin elevates blood pressure and enhances cognitive performance in the elderly population. Eighty women aged between 50 and 80 years were examined based on a randomized, placebo controlled double blind design. Blood pressure was measured sphygmomanometrically and through continuous noninvasive recording. Cognitive performance was assessed by means of two tests measuring general information processing capacity and visuomotor speed. The administration of the drug led to a short term increase in blood pressure as well as in cognitive performance as compared to placebo. Potential physiological mechanisms of action mediating this effect, including hemodynamic alterations, sympathetic stimulation as well as improvement of cerebral metabolism are discussed. PMID- 19560328 TI - No association between the PPARG gene and schizophrenia in a British population. AB - It has consistently been reported that patients with schizophrenia have an increased risk of type-2 diabetes. To investigate a genetic link between these two diseases, the combined effects of the PLA2G4A, PTGS2 and PPARG genes were tested among 221 British nuclear families consisting of fathers, mothers and affected offspring with schizophrenia. A total of 10 single nucleotide polymorphisms (SNPs) were tested and the likelihood-based association analysis for nuclear families was used to analyse the genotyping data. Eight SNPs detected across the PPARG gene did not show allelic association with schizophrenia; a weak association was detected at rs2745557 in the PTGS2 locus (chi2=4.19, p=0.041) and rs10798059 in the PLA2G4A locus (chi2=4.28, p=0.039) but these associations did not survive after 10,000 permutations to correct the p-value (global p=0.246). The gene-gene interaction test did not show any evidence of either cis-phase interactions for the PLA2G4A and PTGS2 combinations or a trans-phase interaction for the PLA2G4A and PPARG combinations. The PPARG gene has been reported to be strongly associated with type-2 diabetes, but the present study did not support the hypothesis that the PPARG gene may also play an important role in the development of schizophrenia. PMID- 19560329 TI - Long-chain n-3 fatty acid levels in baseline serum phospholipids do not predict later occurrence of depressive episodes: a nested case-control study within a cohort of middle-aged French men and women. AB - The goal of this study was to seek the relations between baseline n-3 PUFA status and the later occurrence of depressive episodes in a French cohort of middle-aged men and women, the SU.VI.MAX study. A nested case-control study was designed within the cohort: cases with at least two depressive episodes during the 8-year follow-up were paired to non-depressed controls, antidepressant prescriptions being taken as markers of depressive episodes. The fatty acid profiles of baseline serum phospholipids have been determined. Results were analyzed using logistic regression and principal component analysis, taking into account depression history and demographic and lifestyle confounders. There was no consistent association of depression risk with any serum fatty acid, and in particular there was no association of depression risk with the long-chain n-3 PUFA eicosapentaenoic, docosapentaenoic and docosahexaenoic acids. This study does not support the hypothesis of a predictive value of n-3 PUFA status for depression in population settings. PMID- 19560330 TI - Depression and its relationship with poor exercise capacity, BODE index and muscle wasting in COPD. AB - BACKGROUND: The prevalence of depression in stable COPD patients varies markedly, possibly because of use of different scales. We aimed to assess depression using 2 different depression scales and to examine the association between depression and poor exercise performance, BODE index and muscle wasting in clinically stable COPD patients. METHODS: 122 stable COPD patients were assessed with the Centre for Epidemiologic Studies Depression Scale (CES-D) and the Brief Assessment Schedule Depression Cards (BASDEC). We also assessed patients with spirometry, bioelectrical impedance analysis, 6-minute walk distance (6MWD), St George's Respiratory Questionnaire (SGRQ) and MRC dyspnoea and Borg scales. RESULTS: The CES-D and BASDEC scales detected almost similar prevalence rates of depression (21% vs 17%) with a Kappa coefficient of 0.68, p<0.0001. The BASDEC scale detected more depression in women and was more closely associated with dyspnoea than the CES-D. COPD severity was associated with depression when using BODE scores but not when GOLD categories were used. Each of the CES-D and BASDEC depression scores were associated with 6MWD after adjusting for FEV1% predicted, gender, age and pack-years (p = <0.0001 and 0.001, respectively). Also, patients with a 6MWD<350 scored significantly higher on both depression scales. Wasted patients appeared to have higher depression scores, but the difference was statistically insignificant. CONCLUSION: The administration of different depression scales may affect some of the characteristics of depressed patients rather than the prevalence rate of depression. Depression was associated with poor exercise performance and BODE index in COPD. PMID- 19560331 TI - How ILK and kindlins cooperate to orchestrate integrin signaling. AB - Integrin-mediated cell adhesion regulates multiple cellular processes crucial for development, physiology, and pathology. Since integrins lack enzymatic activity they need to recruit adaptor and signaling proteins to mediate their functions. The cytoplasmic proteins kindlins and integrin-linked kinase (ILK) associate with integrin tails and thereby link integrins with the actin cytoskeleton and various signaling pathways. In comparison to their role in regulating integrin function in cell-matrix adhesions, less is known about the functions of kindlins and ILK in other cellular compartments, such as cell-cell contacts and in the nucleus. PMID- 19560333 TI - Editors' introduction: policy change and policy analysis. PMID- 19560332 TI - Biotin increases glucokinase expression via soluble guanylate cyclase/protein kinase G, adenosine triphosphate production and autocrine action of insulin in pancreatic rat islets. AB - Besides its role as a carboxylase prosthetic group, biotin has important effects on gene expression. However, the molecular mechanisms through which biotin exerts these effects are largely unknown. We previously found that biotin increases pancreatic glucokinase expression. We have now explored the mechanisms underlying this effect. Pancreatic islets from Wistar rats were treated with biotin, in the presence or absence of different types of inhibitors. Glucokinase mRNA and 18s rRNA abundance were determined by real-time PCR. Adenosine triphosphate (ATP) content was analyzed by fluorometry. Biotin treatment increased glucokinase mRNA abundance approximately one fold after 2 h; the effect was sustained up to 24 h. Inhibition of soluble guanylate cyclase or protein kinase G (PKG) signalling suppressed biotin-induced glucokinase expression. The cascade of events downstream of PKG in biotin-mediated gene transcription is not known. We found that inhibition of insulin secretion with diazoxide or nifedipine prevented biotin-stimulated glucokinase mRNA increase. Biotin treatment increased islet ATP content (control: 4.68+/-0.28; biotin treated: 6.62+/-0.26 pmol/islet) at 30 min. Inhibition of PKG activity suppressed the effects of biotin on ATP content. Insulin antibodies or inhibitors of phosphoinositol-3-kinase/Akt insulin signalling pathway prevented biotin-induced glucokinase expression. The nucleotide 8-Br-cGMP mimicked the biotin effects. We propose that the induction of pancreatic glucokinase mRNA by biotin involves guanylate cyclase and PKG activation, which leads to an increase in ATP content. This induces insulin secretion via ATP-sensitive potassium channels. Autocrine insulin, in turn, activates phosphoinositol-3-kinase/Akt signalling. Our results offer new insights into the pathways that participate in biotin-mediated gene expression. PMID- 19560334 TI - Effect of industrial by-products containing electron acceptors on mitigating methane emission during rice cultivation. AB - Three industrial by-products (fly ash, phosphogypsum and blast furnace slag), were evaluated for their potential re-use as soil amendments to reduce methane (CH(4)) emission resulting from rice cultivation. In laboratory incubations, CH(4) production rates from anoxic soil slurries were significantly reduced at amendment levels of 0.5%, 1%, 2% and 5% (wt wt(-1)), while observed CO(2) production rates were enhanced. The level of suppression in methane production was the highest for phosphogypsum, followed by blast slag and then fly ash. In the greenhouse experiment, CH(4) emission rates from the rice planted potted soils significantly decreased with the increasing levels (2-20 Mg ha(-1)) of the selected amendments applied, while rice yield simultaneously increased compared to the control treatment. At 10 Mg ha(-1) application level of the amendments, total seasonal CH(4) emissions were reduced by 20%, 27% and 25%, while rice grain yields were increased by 17%, 15% and 23% over the control with fly ash, phosphogypsum, and blast slag amendments, respectively. The suppression of CH(4) production rates as well as total seasonal CH(4) flux could be due to the increased concentrations of active iron, free iron, manganese oxides, and sulfate in the amended soil, which acted as electron acceptors and controlled methanogens' activity by limiting substrates availability. Among the amendments, blast furnace slag and fly ash contributed mainly to improve the soil nutrients balance and increased the soil pH level towards neutral point, but soil acidity was developed with phosphogypsum application. Conclusively, blast slag among the selected amendments would be a suitable soil amendment for reducing CH(4) emissions as well as sustaining rice productivity. PMID- 19560335 TI - Electrochemical analysis of copper ion using a Gly-Gly-His tripeptide modified poly(3-thiopheneacetic acid) biosensor. AB - A novel biosensor harnessing a conducting polymer functionalized with a copper ion specific peptide proved to be highly effective for electrochemical analysis of copper ions. The developed sensor comprised a transducer based on a conducting polymer (poly(3-thiopheneacetic acid)) electrode and a probe (tripeptide, Gly-Gly His) selectively cognitive of copper ions. For functionalization of the electrode, the carboxylic group of the polymer was covalently coupled with the amine group of the tripeptide, and its structural features were confirmed by X ray photoelectron spectroscopy (XPS) and attenuated total reflection infrared (ATR-IR) spectroscopy. The peptide modified polythiophene biosensor was used for the electrochemical analysis of various trace metal ions by square wave voltammetry. The electrode was found to be highly sensitive and selective to Cu(2+) in the range of 0.02-20 microM with almost no cross binding to other metal ions such as Ni(2+) and Pb(2+). Furthermore, the developed sensor exhibited a high stability and reproducibility despite the repeated use of the sensor electrode and probe. With the advent of more diverse affinity bioprobes specific towards a broad range of analytes, the demonstrated strategy harnessing peptide modified polythiophene biosensor is likely to provide an excellent platform for the selective determination of trace amount of analytes whose detection is otherwise cumbersome. PMID- 19560337 TI - We march backwards into the future. PMID- 19560336 TI - Engineering algae for biohydrogen and biofuel production. AB - There is currently substantial interest in utilizing eukaryotic algae for the renewable production of several bioenergy carriers, including starches for alcohols, lipids for diesel fuel surrogates, and H2 for fuel cells. Relative to terrestrial biofuel feedstocks, algae can convert solar energy into fuels at higher photosynthetic efficiencies, and can thrive in salt water systems. Recently, there has been considerable progress in identifying relevant bioenergy genes and pathways in microalgae, and powerful genetic techniques have been developed to engineer some strains via the targeted disruption of endogenous genes and/or transgene expression. Collectively, the progress that has been realized in these areas is rapidly advancing our ability to genetically optimize the production of targeted biofuels. PMID- 19560338 TI - Biological kinetics evaluation of anaerobic stabilization pond treatment of palm oil mill effluent. AB - Biological kinetic (bio-kinetic) study of the anaerobic stabilization pond treatment of palm oil mill effluent (POME) was carried out in a laboratory anaerobic bench scale reactor (ABSR). The reactor was operated at different feed flow-rates of 0.63, 0.76, 0.95, 1.27, 1.9 and 3.8l of raw POME for a day. Chemical oxygen demand (COD) as influent substrates was selected for bio-kinetic study. The investigation showed that the growth yield (Y(G)), specific biomass decay (b), maximum specific biomass growth rate (mu(max)), saturation constant (K(s)) and critical retention time (Theta(c)) were in the range of 0.990 g VSS/g COD(removed) day, 0.024 day(-1), 0.524 day(-1), 203.433 g COD l(-1) and 1.908 day, respectively. PMID- 19560339 TI - Production of an acidic and thermostable lipase of the mesophilic fungus Penicillium simplicissimum by solid-state fermentation. AB - The production of a lipase by a wild-type Brazilian strain of Penicillium simplicissimum in solid-state fermentation of babassu cake, an abundant residue of the oil industry, was studied. The enzyme production reached about 90 U/g in 72 h, with a specific activity of 4.5 U/mg of total proteins. The crude lipase showed high activities at 35-60 degrees C and pH 4.0-6.0, with a maximum activity at 50 degrees C and pH 4.0-5.0. Enzyme stability was enhanced at pH 5.0 and 6.0, with a maximum half-life of 5.02 h at 50 degrees C and pH 5.0. Thus, this lipase shows a thermophilic and thermostable behavior, what is not common among lipases from mesophilic filamentous fungi. The crude enzyme catalysed the hydrolysis of triglycerides and p-nitrophenyl esters (C4:0-C18:0), preferably acting on substrates with medium-chain fatty acids. This non-purified lipase in addition to interesting properties showed a reduced production cost making feasible its applicability in many fields. PMID- 19560340 TI - Efficient metal removal and neutralization of acid mine drainage by crab-shell chitin under batch and continuous-flow conditions. AB - Crab-shell chitin was evaluated as a multifunctional substrate for treating acid mine drainage (AMD) in both batch-microcosms and continuous-flow column tests. In microcosms, crab-shell chitin was able to treat AMD from three different sites with similar results: pH increased from 3.5 to approximately 7.5 within 2 days; alkalinity increased at a rate of 37.9+/-2.2 mg CaCO(3)/L day; and sulfate was reduced at a rate of -13.6+/-2.6 mg SO(4)(2-)/L day. In columns, a hydraulic retention time of 11.2 h was enough to raise the pH from 3.5 to approximately 7.5. Alkalinity increased at a rate of 50+/-20 mg CaCO(3)/day, and lasted throughout the duration of the test (125 days, 268 pore volumes (PV)) without showing signs of exhaustion. Metals (Al, Fe, and Mn) were completely removed for 171 PV, and geochemical modeling indicates that they likely precipitated as insoluble hydr(oxides), sulfides, and carbonates. Manganese and iron breakthroughs occurred after 174 and 234 PV, respectively, whereas aluminum breakthrough was never observed. These results demonstrate for the first time that crab-shell chitin can completely remove metals and neutralize the pH of AMD under continuous-flow conditions. PMID- 19560341 TI - Effect of fiber treatments on tensile and thermal properties of starch/ethylene vinyl alcohol copolymers/coir biocomposites. AB - Coir fibers received three treatments, namely washing with water, alkali treatment (mercerization) and bleaching. Treated fibers were incorporated in starch/ethylene vinyl alcohol copolymers (EVOH) blends. Mechanical and thermal properties of starch/EVOH/coir biocomposites were evaluated. Fiber morphology and the fiber/matrix interface were further characterized by scanning electron microscopy (SEM). All treatments produced surface modifications and improved the thermal stability of the fibers and consequently of the composites. The best results were obtained for mercerized fibers where the tensile strength was increased by about 53% as compared to the composites with untreated fibers, and about 33.3% as compared to the composites without fibers. The mercerization improved fiber-matrix adhesion, allowing an efficient stress transfer from the matrix to the fibers. The increased adhesion between fiber and matrix was also observed by SEM. Treatment with water also improved values of Young's modulus which were increased by about 75% as compared to the blends without the fibers. Thus, starch/EVOH blends reinforced with the treated fibers exhibited superior properties than neat starch/EVOH. PMID- 19560343 TI - Improvement of yield of Pleurotus eryngii var. eryngii by substrate supplementation and use of a casing overlay. AB - Improved yield and biological efficiency (BE) of Pleurotus eryngii var. eryngii were achieved by supplementation of substrate with a commercial delayed-release nutrient and use of a casing overlay. Yield increases of 14% were achieved from cased substrates that were supplemented at time of casing with delayed-release nutrient (Remo's). Use of a casing layer enhanced yield by 141% over non-cased substrates. When casing and substrate supplementation were combined, yield increased 179% over non-cased/non-supplemented substrates. Mushrooms harvested from cased substrates were darker in color and solids contents were lower compared to non-cased substrates. An additional break of mushrooms was harvested from non-cased "spent" substrate by fragmenting and re-supplementing the substrate prior to the application of a casing overlay. Three production methods were compared for their effect on mushroom yield: "standard", "casing" and "casing after first break". Casing of the substrate before first break ("casing" production method) resulted in the highest yield and biological efficiency. PMID- 19560342 TI - Prediction of biogas potentials using quick laboratory analyses: upgrading previous models for application to heterogeneous organic matrices. AB - This study presents an upgrading of the mathematical models to predict anaerobic biogasification potential (ABP) through quick laboratory analyses that have been presented in an earlier study. The aim is to widen the applicability of the models to heterogeneous organic substrates and to improve their reliability through a deeper statistical approach. Three multiple-step linear regressions were obtained using biomass oxygen demand in 20 h (OD(20)) plus the volatile solids content (VS) of 23 new samples of heterogeneous organic matrices, of 46 samples presented in the earlier work and of the data set comprising all the 69 samples. The two variables chosen were found to be suitable for very heterogeneous materials. To judge the prediction quality, a validation procedure was performed with 12 new samples using model efficiency indexes. The proposed model had good prediction ability for a large variety of organic substrates, and allows the calculation of the ABP value within only 2-day's laboratory work instead of the 60-90 days required to obtain ABP by anaerobic test. PMID- 19560344 TI - Lessons from the cow: what the ruminant animal can teach us about consolidated bioprocessing of cellulosic biomass. AB - Consolidated bioprocessing (CBP) of cellulosic biomass is a promising source of ethanol. This process uses anaerobic bacteria, their own cellulolytic enzymes and fermentation pathways that convert the products of cellulose hydrolysis to ethanol in a single reactor. However, the engineering and economics of the process remain questionable. The ruminal fermentation is a very highly developed natural cellulose-degrading system. We propose that breakthroughs developed by cattle and other ruminant animals in cellulosic biomass conversion can guide future improvements in engineered CBP systems. These breakthroughs include, among others, an elegant and effective physical pretreatment; operation at high solids loading under non-aseptic conditions; minimal nutrient requirements beyond the plant biomass itself; efficient fermentation of nearly all plant components; efficient recovery of primary fermentation end-products; and production of useful co-products. Ruminal fermentation does not produce significant amounts of ethanol, but it produces volatile fatty acids and methane at a rapid rate. Because these alternative products have a high energy content, efforts should be made to recover these products and convert them to other organic compounds, particularly transportation fuels. PMID- 19560345 TI - Evaluation of chromium(VI) removal behaviour by two isolates of Synechocystis sp. in terms of exopolysaccharide (EPS) production and monomer composition. AB - Chromium(VI) removal and its association with exopolysaccharide (EPS) production in cyanobacteria were investigated. Synechocystis sp. BASO670 produced higher EPS (548 mg L(-1)) than Synechocystis sp. BASO672 (356 mg L(-1)). While the EC(50) of the Cr(VI) for Synechocystis sp. BASO670 and Synechocystis sp. BASO672 were determined as 11.5 mg L(-1), and 2.0 mg L(-1), respectively, there was no relation between Cr(VI) removal and EPS production. Synechocystis sp. BASO672, which has higher EPS value, removed (33%) more Cr(VI) than Synechocystis sp. BASO670. Monomer compositions of EPS of each of the isolates were determined differently. Synechocystis sp. BASO672 which removed higher Cr(VI), had higher values of uronic acid and glucuronic acid (192 microg/mg and 89%, respectively). Our results showed that EPS might play a role in Cr(VI) tolerance. Monomer composition, especially uronic acid and glucuronic acid content of EPS may have enhanced Cr(VI) removal. PMID- 19560346 TI - Roles of polyurethane foam in aerobic moving and fixed bed bioreactors. AB - The aim of this study was to investigate the performance of sponge as an active mobile carrier for attached-growth biomass in three typical types of aerobic bioreactors to treat a high strength synthetic wastewater. The results show that sponge thickness deteriorated the organic and nutrient removal and 1cm is the optimum thickness for fixed-bed sponge biofilter (SBF). The sponge volume had significant impact on phosphorus removal rather than organic or nitrogen removal, and 20% volume of sponge could achieve 100% T-P removal within 3h in a sponge batch reactor (SBR). When sponge coupled with submerged membrane bioreactor (SMBR), the single system show outstanding ammonium (100% at filtration flux of 10 and 15 L/m(2)h) and phosphorus (> 91% at all fluxes range) removal with optimum pH range of 6-7. PMID- 19560347 TI - Effect of anatomical fractionation on the enzymatic hydrolysis of acid and alkaline pretreated corn stover. AB - Due to concerns with biomass collection systems and soil sustainability there are opportunities to investigate the optimal plant fractions to collect for conversion. An ideal feedstock would require a low severity pretreatment to release a maximum amount of sugar during enzymatic hydrolysis. Corn stover fractions were separated manually and analyzed for glucan, xylan, acid soluble lignin, acid insoluble lignin, and ash composition. The stover fractions were also pretreated with either 0%, 0.4%, or 0.8% NaOH for 2 h at room temperature, washed, autoclaved and saccharified. In addition, dilute sulfuric acid pretreated samples underwent simultaneous saccharification and fermentation (SSF) to ethanol. In general, the two pretreatments produced similar trends with cobs, husks, and leaves responding best to the pretreatments, the tops of stalks responding slightly less, and the bottom of the stalks responding the least. For example, corn husks pretreated with 0.8% NaOH released over 90% (standard error of 3.8%) of the available glucan, while only 45% (standard error of 1.1%) of the glucan was produced from identically treated stalk bottoms. Estimates of the theoretical ethanol yield using acid pretreatment followed by SSF were 65% (standard error of 15.9%) for husks and 29% (standard error of 1.8%) for stalk bottoms. This suggests that integration of biomass collection systems to remove sustainable feedstocks could be integrated with the processes within a biorefinery to minimize overall ethanol production costs. PMID- 19560348 TI - Influence of bulking agent on sewage sludge composting process. AB - Four types of compost, consisting of mixtures of Acacia dealbata (A) with sewage sludge (SS) were studied in a laboratory reactor. Composting time was 80 days and parameters monitored over this period included temperature, organic matter, pH, CO(2), O(2), C/N ratio, Kjeldahl-N, as well as maturity indexes. All the studied parameters were influenced by the bulking amount used. The highest profile temperature measured was for the A/SS 1/2 (w/w) mixture that reached a maxima temperature of 67 degrees C and lower maximum temperatures of 52, 48 and 46 degrees C were observed for A/SS 1/3, 1/1 and 1/0 composts, respectively. The kinetic model used showed that a descent of sewage sludge in the composting mixtures favored the enzyme-substrate affinity. However, an increase in depending on the parameters of the process factors was observed when the sewage sludge ratio was increased in mixtures. The optimal amounts of sewage sludge for co composting with Acacia indicate that moderate amounts of sludge (1/1) would be the best compromise. PMID- 19560349 TI - Investigation of biomass and lipids production with Neochloris oleoabundans in photobioreactor. AB - The fresh water microalga Neochloris oleoabundans was investigated for its ability to accumulate lipids and especially triacylglycerols (TAG). A systematic study was conducted, from the determination of the growth medium to its characterization in an airlift photobioreactor. Without nutrient limitation, a maximal biomass areal productivity of 16.5 g m(-2) day(-1) was found. Effects of nitrogen starvation to induce lipids accumulation was next investigated. Due to initial N. oleoabundans total lipids high content (23% of dry weight), highest productivity was obtained without mineral limitation with a maximal total lipids productivity of 3.8 g m(-2) day(-1). Regarding TAG, an almost similar productivity was found whatever the protocol was: continuous production without mineral limitation (0.5 g m(-2) day(-1)) or batch production with either sudden or progressive nitrogen deprivation (0.7 g m(-2) day(-1)). The decrease in growth rate reduces the benefit of the important lipids and TAG accumulation as obtained in nitrogen starvation (37% and 18% of dry weight, respectively). PMID- 19560351 TI - Esters of betulin and betulinic acid with amino acids have improved water solubility and are selectively cytotoxic toward cancer cells. AB - Betulin and betulinic acid are naturally occurring pentacyclic triterpenes showing cytotoxicity towards a number of cancer cell lines. Unfortunately they are practically insoluble in aqueous media and therefore their overall absorption index is not satisfactory. We have modified structures of both compounds by simple transformation to mono- and disubstituted esters of l-amino acids. This allowed us to achieve better water solubility without loss of the observed earlier anticancer properties. Comet assay on various cancer cell lines demonstrate that these compounds act via an apoptotic mechanism. PMID- 19560350 TI - Synthesis of novel macrolide derivatives with imidazo[4,5-b]pyridinyl sulfur contained alkyl side chains and their antibacterial activity. AB - In an effort to find new antibiotics, a novel series of 14-membered macrolides with imidazo[4,5-b]pyridinyl sulfur contained alkyl side chains has been synthesized based on commercially available clarithromycin. Chemical transformation of hydroxy group at position C-3 afforded range of ketolides and acylides. Compared to telithromycin, compound 15a demonstrated improved in vitro activity against erythromycin-susceptible and -resistant strains. PMID- 19560352 TI - Synthesis and application of photoaffinity probe containing an intact isoprenoid chain. AB - Two novel chemical probes each carrying an intact isoprenoid chain, a biotin tag and a benzophenone moiety were synthesized. Photoaffinity labeling of the Saccharomyces cerevisiae cell lysate revealed that these probes could selectively trap some proteins, and proteins with molecular weight of approximately 70 KDa appeared as a major band upon Streptavidin blot analysis. PMID- 19560353 TI - Identification of new Hsp90 inhibitors by structure-based virtual screening. AB - Structure-based virtual screening identified pyrimidine-2,4,6-trione and 4H-1,2,4 triazole-3-thiol as novel scaffolds of Hsp90 ATPase inhibitors. Their binding modes in the ATP-binding pocket of Hsp90 were analyzed using AutoDoc program combined with molecular dynamics (MD) simulations. PMID- 19560355 TI - Discovery of a new class of catalytic topoisomerase II inhibitors targeting the ATP-binding site by structure based design. Part I. AB - Topoisomerase II is a validated target in oncology. Among the different ways of blocking the function of this enzyme, inhibiting its ATPase activity has been relatively less investigated. In an effort to identify topoisomerase II inhibitors of a novel type, exerting their action by this mechanism, we have designed a purine inhibitor scaffold targeting the ATP-binding site of the enzyme. Searching the Novartis compound collection for molecules containing this purine motif has allowed the identification of two micromolar hits providing access to a new class of catalytic topoisomerase II inhibitors. PMID- 19560354 TI - Evaluation of structurally diverse neuronal nicotinic receptor ligands for selectivity at the alpha6( *) subtype. AB - Direct comparison of pyridine versus pyrimidine substituents on a small but diverse set of ligands indicates that the pyrimidine substitution has the potential to enhance affinity and/or functional activity at alpha6 subunit containing neuronal nicotinic receptors (NNRs) and decrease activation of ganglionic nicotinic receptors, depending on the scaffold. The ramifications of this structure-activity relationship are discussed in the context of the design of small molecules targeting smoking cessation. PMID- 19560356 TI - Developing standard procedures for murine and canine efficacy studies of DMD therapeutics: report of two expert workshops on "Pre-clinical testing for Duchenne dystrophy": Washington DC, October 27th-28th 2007 and Zurich, June 30th July 1st 2008. PMID- 19560357 TI - Intraflagellar transport and the generation of dynamic, structurally and functionally diverse cilia. AB - Cilia are organelles that project from most eukaryotic organisms and cell types. Their pervasiveness stems from having remarkably versatile propulsive and sensory functions, which in humans are recognized to have essential roles in physiology and development. Under-appreciated, however, are their diverse ultrastructures and typically bipartite organization consisting of doublet and singlet microtubules. Moreover, the overall shapes of the membrane-ensheathed cilia are varied, as exemplified by differences between hair-like olfactory cilia and rod- or cone-shaped photoreceptor connecting cilia-outer segments. Although cell specific transcriptional programs are evidently crucial in establishing ciliary morphological specialization, few players directly involved in generating such diversity are known. Recent findings suggest that at least two molecular motors (kinesin-II and OSM-3/KIF17) can differentially mobilize the intraflagellar transport machinery required for ciliogenesis and, presumably, different cargo to help generate dynamic, structurally and functionally distinct cilia. PMID- 19560359 TI - Aberrant pelvis and hip kinematics impair hip loading before and after total hip replacement. AB - Musculoskeletal loading is an important factor affecting the development of osteoarthritis, bone remodelling and primary fixation of total hip replacement (THR). In this study, we analyzed the relation between muscular force, gait kinematics and kinetics and hip loading in 20 patients before and six weeks after THR. Hip contact forces were calculated from gait analysis data using musculoskeletal modelling, inverse dynamics and static optimization. We found aberrant pelvis and hip kinematics and kinetics before and six weeks after surgery, confirming previous findings in literature. Furthermore, we found a decrease in the total contact force and its vertical component. These changes result in a decrease of the associated inclination angles of the total hip contact force in the sagittal and transverse planes, changing the orientation towards more vertical implant loading after THR. These changes in hip loading were related to observed gait kinematics and kinetics. Most importantly, excessive pelvic obliquity and associated hip adduction related to impaired implant loading. We concluded, therefore, that physical therapy in the early post operative phase should primarily focus on stretching of anterior and medial structures and strengthening of hip flexors and abductors to achieve normalization of the hip and pelvis kinematics and consequently normalize hip loading. PMID- 19560360 TI - Visually evoked whole-body turning responses during stepping in place in a virtual environment. AB - Humans use a specific sequence of reorientation of the eyes, head and body to perform turning and redirections while walking. Gaze (eye and head) rotation in a new direction of travel precedes body rotation by as much as 1.5s and provides a stable reference frame that guides subsequent whole-body redirection. The purpose of the current study was to determine whether a visually presented rotation of the external environment can induce whole-body turning lead by gaze redirection in a new travel direction. Five healthy young adults performed a stepping in place task while watching a virtual scene that moved as if they were walking down a hallway, thus providing participants with a perception of forward self motion. While "forward" stepping, the virtual scene would gradually turn around a 90 degrees corner. As a result the turn could be anticipated by the participants. Significant horizontal eye movements and head and body rotation magnitudes were observed in response to the virtual visual turning cue. Onset of eye, head and body redirection revealed a sequenced order and timing of segment rotation that is characteristic of steering behaviour in real world turning situations. The results of this study provide support for the hypothesis that gaze redirection may be an essential subcomponent to steering behaviour. The link between visual redirection and coordinated body turning implies instability when turning may result from visual and/or oculomotor deficits. PMID- 19560358 TI - A quest for the mechanism regulating global planar cell polarity of tissues. AB - Most epithelial cells, besides their ubiquitous apical-basal polarity, are polarized within the plane of the epithelium, which is called planar cell polarity (PCP). Using Drosophila as a model, meaningful progress has been made in the identification of key PCP factors and the dissection of their intracellular molecular interactions. The long-range, global aspects of coordinated polarization and the overlying regulatory mechanisms that create the initial polarity direction have, however, remained elusive. Several recent publications have outlined potential mechanisms of how the global regulation of PCP might be controlled and how the distinct core factor groups might interact via frizzled, Van Gogh or flamingo. This review focuses on these exciting features and attempts to provide an integrated picture of these recent and novel insights. PMID- 19560361 TI - Modified orbitozygomatic craniotomy for large medial sphenoid wing meningiomas. AB - Modified orbitozygomatic craniotomy (MOZC) is an anterior lateral skull base approach characterized by simplicity and wide exposure. The approach was first introduced in 2003 and there are few clinical reports. This report details treatment of patients with large (>4 cm) sphenoid wing meningiomas via a MOZC approach, and to the authors' knowledge, the first published in English. Total resection was achieved in all 5 patients in this study. One patient experienced a postoperative epidural hematoma that was successfully treated. All patients returned to daily activity without neurological sequellae. The advantages of MOZC are sparing of the zygomatic arch and removal of the orbital rim; hence, the surgeon can plan a capacious operative field without excessive brain retraction and resect the tumor before opening the dura. The MOZC approach is a clinically feasible, low morbidity, surgical option for paraclinoid lesions, such as large sphenoid wing meningiomas. PMID- 19560362 TI - Regression of intracranial meningioma following intratumoral hemorrhage. AB - An intratumoral hemorrhage within a meningioma occurs infrequently; in less than 3% of all lesions. When hemorrhage does occur, however, it is associated with a poor prognosis and significant mortality rates. We report a 66-year-old woman with a 10-year history of multiple intracranial meningiomas managed conservatively who underwent surgical resection of a spheno-orbital lesion for decompression of the right optic nerve. Postoperatively, an intratumoral hemorrhage developed in a contralateral lesion, which was managed conservatively. During follow up, the hemorrhaged lesion became significantly smaller. To our knowledge there are no published reports of spontaneous resolution of a meningioma after intratumoral hemorrhage without surgical management. We review the literature on hemorrhage in meningiomas and postulate some pathophysiologic mechanisms for the bleeding and subsequent tumor resolution seen in this patient. PMID- 19560364 TI - Characterization and synthetic application of a novel beta1,3 galactosyltransferase from Escherichia coli O55:H7. AB - A beta1,3-galactosyltransferase (WbgO) was identified in Escherichia coli O55:H7. Its function was confirmed by radioactive activity assay and structure analysis of the disaccharide synthesized with the recombinant enzyme. WbgO requires a divalent metal ion, either Mn(2+) or Mg(2+), for its activity and is active between pH 6.0-8.0 with a pH optimum of 7.0. N-acetylglucosamine (GlcNAc) and oligosaccharides with GlcNAc at the non-reducing end were shown to be its preferred substrates and it can be used for the synthesis of type 1 glycan chains from these substrates. Together with a recombinant bacterial GlcNAc-transferase, benzyl beta-lacto-N-tetraoside was synthesized with the purified WbgO to demonstrate the synthetic utility of WbgO. PMID- 19560363 TI - In vitro biological activity and structural analysis of 2,4-diamino-5-(2' arylpropargyl)pyrimidine inhibitors of Candida albicans. AB - In order to develop new antifungal agents effective against two species of Candida, we have designed a series of dihydrofolate reductase (DHFR) inhibitors. Here, we explore the structure-activity relationships of these inhibitors toward Candida albicans DHFR by evaluating enzyme inhibition, antifungal activity and toxicity to mammalian cells. Analysis of docked complexes of the enzyme and inhibitors yields the structural basis of relative potency. The meta-biphenyl series of this class exhibits the greatest enzyme inhibition, selectivity and antifungal activity. PMID- 19560365 TI - Effects of static magnetic fields on Escherichia coli. AB - This paper represents the study of the biological effects of static magnetic fields (SMFs) on Escherichia coli (E. coli). The bacterial strain E. coli was exposed to SMFs in order to test its viability (evaluated by the number of colony forming units (CFU)). In this study, we measured the dependence of CFU on the duration of exposure, on the treatment temperature T and on the value of the magnetic field induction B. The results showed that the number of CFU decreased with longer exposure time and higher treatment temperature (from 25 degrees C to 40 degrees C), whereas multiple extreme values of number of CFU were obtained when the induction B changed. In order to explain the results, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were used to investigate the morphology of the cells. We found obvious cell surface damage when the cells were exposed to SMFs. PMID- 19560366 TI - A simplified surgical approach for aortic valve replacement after previous coronary artery bypass grafting. AB - Aortic valve replacement (AVR) in patients who have undergone previous coronary artery bypass grafting (CABG) is a challenging redo surgery. We undertook this study to evaluate the early and late outcomes of patients operated upon using a simplified surgical approach. Between January 2001 and December 2005, 2238 patients underwent AVR in our institution. We reviewed retrospectively the 57 patients who had AVR following previous CABG. All patients underwent cardiopulmonary bypass with a mild-to-moderate systemic hypothermia (mean temperature: 29.7 +/- 2.5 degrees C). Patent internal thoracic artery (IMA) grafts were never dissected, controlled or clamped. A mechanical or biological prosthesis was implanted considering the patient's age. The mean cardiopulmonary bypass (CPB) time was 93 +/- 29 min (median: 80 min, range: 43-244 min) and the mean aortic cross-clamp (AoX) time was 63 +/- 18 min (median: 59 min, range: 31 125 min). The early mortality was 10.5% and the late mortality was 9.8% (mean follow-up time: 38 months). The survival was 81% at 5 years and the freedom from major cardiac events was 77%. In conclusion, from our experience, the operating quickness and a simplified approach ('open IMA technique', anterograde cardioplegia, mild-to-moderate hypothermia and minimal dissection of the mediastinal structures) represent two fundamental choices to perform this type of surgery easily, safely and with optimal results. PMID- 19560367 TI - Comparation of liposomal formulations of ALA Undecanoyl ester for its use in photodynamic therapy. AB - ALA administration has been used to induce the endogenous photosensitiser Protoporphyrin IX for photodynamic therapy (PDT) of tumours. However, the hydrophilic nature of ALA limits its ability to penetrate through skin restricting the use of ALA-PDT to superficial diseases. Lipophilic derivatives of ALA such as ALA Undecanoyl ester (Und-ALA) were designed to have better diffusing properties. However, Und-ALA, applied topically on the skin over the tumour, induced low porphyrin content. To improve Und-ALA efficacy we tested the efficacy of Und-ALA as porphyrin inducer, delivered in phosphatidylcholine and phosphatidylglycerol (PC-PG) or phosphatidylcholine and phosphatidic acid (PC-PA) liposomal formulations. Entrapment of Und-ALA into PC-PA or PC-PG liposomes resulted in a dramatic impairment of toxicity in the mammary tumour LM3 cells. However, liposomal Und-ALA induced lower intracellular porphyrin content compared to free ALA, although total porphyrins content (intracellular+media) from free Und-ALA resulted equal compared to liposomal Und-ALA, due to induction of porphyrins release induced by the latter. Topical administration of Und-ALA in PC PG or PC-PA liposomes over the skin of LM3 subcutaneously injected mice, induced equal amount of tumour porphyrins as compared to free Und-ALA. The kinetics of porphyrins synthesis from Und-ALA is similar for free and liposomal formulations both in vivo and in vitro, showing that release of Und-ALA from liposomes is not gradual and suggesting that liposome membranes either fuses or binds to the cell membranes. To sum up, the incorporation of Und-ALA into liposomes of PC-PA or PC PG composition does not improve the rate of porphyrin synthesis either in vitro or in vivo, due to a massive release of extracellular porphyrins and a poor cytoplasmatic release of the liposome content. The design of new liposome compositions either favouring endocytosis or coated with natural polymers to prevent Und-ALA interaction with cellular membrane are desired to overcome intracellular porphyrin release after long-chained ALA esters treatment. PMID- 19560368 TI - Adjuvant chemotherapy in elderly patients with colorectal cancer. A retrospective analysis of the implementation of tumor board recommendations in a single institution. AB - BACKGROUND: A number of studies have shown that elderly cancer patients were denied optimal anticancer treatment because of age. Colorectal cancer is among the most frequent cancers in Western countries, and adjuvant chemotherapy has proven efficacy and tolerance in this condition. This study was undertaken to explore the current approaches to adjuvant chemotherapy in elderly cancer patients in a single institution. PATIENTS AND METHODS: We retrospectively analyzed all patients' files that were discussed in the gastro-intestinal tumor board of the Hopitaux Universitaires de Strasbourg during 3 years (2004-2006). The recorded variables included sex, age, tumor stage, cancer location colon vs rectum, number of comorbidities, occurrence of an oncogeriatric assessment, type and tolerance of chemotherapy. We investigated the reason to not administer adjuvant therapy in patients whom should have received this treatment if guidelines had to be applied. RESULTS: A total of 193 consecutive patients' files were extracted from colorectal cancer patients that had been discussed in the gastro-intestinal tumor board. Among these, we isolated patients over 70 years old who were proposed with either adjuvant chemotherapy (group A, n=65) or follow up (group B, n=128). The median age in group A was 75.3 years old. Tumor board recommendations were in accordance with guidelines in 91% of cases. Chemotherapy was delivered in 44 pts (76%) and completed in 42 (95%). The median age in group B was 78.6 years old, and in this group tumor board proposal met the guidelines in 83% of cases. In the logistic regression model, disease stage was the major variable leading to adjuvant treatment recommendation, age and comorbidities being of lesser importance. CONCLUSIONS: In our series, elderly colorectal cancer patients are not undertreated. Efforts should be maintained to educate physicians with regard to feasibility of adjuvant chemotherapy in elderly patients. PMID- 19560370 TI - The number of index components affects the diagnostic accuracy of a diet quality index: the role of intracorrelation and intercorrelation structure of the components. AB - PURPOSE: The aim of this work was to evaluate whether the number of components influences the diagnostic accuracy of a diet quality index and whether this association is affected by the intercorrelation structure of components and by the association of components with an outcome (intracorrelation). METHODS: Simulated data were used to develop theoretical indices with various intracorrelation and intercorrelation structures of the components and outcomes. Moreover, dietary intake data of 668 elderly people from the MEDIS (Mediterranean Islands) study were also used to develop a diet index and to test it toward obesity status (outcome). RESULTS: On the basis of 1,000 simulations, we observed that the diagnostic accuracy of an index increases as the number of components increases, only when the components are not intercorrelated or have low intercorrelation. Moreover, the diagnostic accuracy of an index developed with all components associated with an outcome is higher compared with an index developed by using only some components related to the outcome. Finally, the predictive ability of an isolated component is lower than that of an index developed by using non-intercorrelated or low-intercorrelated components. Real data confirmed the aforementioned findings. CONCLUSION: Low-intercorrelated or non-intercorrelated components, strongly associated with a particular outcome, should be used in order to obtain an accurate composite index. PMID- 19560369 TI - Cardiovascular risk factors among Asian Americans: results from a National Health Survey. AB - PURPOSE: We assessed the prevalence of major cardiovascular disease (CVD) risk factors among Chinese, Asian Indian, Filipino, and other Asian populations compared to non-Hispanic Whites in the United States. METHODS: We analyzed aggregated data from the National Health Interview Survey (NHIS) from 2003 to 2005. Bivariate analyses were used to determine differences in the prevalence of CVD risk factors among Asian subgroups and white adults. Logistic regression analyses were also conducted to compare each Asian subgroup with white adults after taking sociodemographic variables into account. RESULTS: The unadjusted prevalence of physical inactivity was highest among Asian Indians and other Asians. After we controlled for covariates, Asian Indians still had higher odds of physical inactivity than Whites (odds ratio [OR]=1.50, 95% confidence interval [CI]=1.22-1.84). All Asian ethnic groups were significantly less likely than Whites to report smoking, obesity, and binge drinking. Compared with Whites, Filipinos were more likely to have hypertension (OR=1.18, 95% CI=1.02-1.44) and Asian Indians were more likely to have diabetes (OR=2.27, 95% CI=1.63-3.20). CONCLUSION: Although Asian race was generally associated with lower risk for CVD, certain risk factors were particularly high among some Asian subgroups. Future interventions should specify the needs of specific subgroups and design culturally specific programs to reduce health risk behaviors in each Asian subpopulation. PMID- 19560371 TI - Tracking and decomposing health and disease inequality in Thailand. AB - PURPOSE: In middle-income countries, interest in the study of inequalities in health has focused on aggregate types of health outcomes, like rates of mortality. This work moves beyond such measures to focus on disease-specific health outcomes with the use of national health survey data. METHODS: Cross sectional data from the national Health and Welfare Survey 2003, covering 52,030 adult aged 15 or older, were analyzed. The health outcomes were the 20 most commonly reported diseases. The age-sex adjusted concentration index (C *) of ill health was used as a measure of socioeconomic health inequality (values ranging from -1 to +1). A negative (or positive) concentration index shows that a disease was more concentrated among the less well off (or better off). Crude concentration indices (C) for four of the most common diseases were also decomposed to quantify determinants of inequalities. RESULTS: Several diseases, such as malaria (C * = -0.462), goiter (C * = -0.352), kidney stone (C * = 0.261), and tuberculosis (C * = -0.233), were strongly concentrated among those with lower incomes, whereas allergic conditions (C * = 0.174) and migraine (C * = 0.085) were disproportionately reported by the better off. Inequalities were found to be associated with older age, low education, and residence in the rural Northeast and rural North of Thailand. CONCLUSIONS: Pro-equity health policy in Thailand and other middle-income countries with health surveys can now be informed by national data combining epidemiological, socioeconomic and health statistics in ways not previously possible. PMID- 19560372 TI - Electromyographic assessment of Gait function following limb salvage procedures for bone sarcoma. AB - Bone sarcomas are the fourth most common cancer in individuals under 25 years of age. Limb salvage procedures have become increasingly popular for the treatment of osteosarcomas as they have functional and physiological benefits over traditional amputative procedures. The purpose of this study was to assess locomotor patterns post lower limb salvage surgery via electromyographic and energetic measurement techniques on a group of intra-articular knee osteosarcoma patients greater than one year post surgery. A retrospective outcome study was undertaken on 20 limb salvage patients (10 female, 10 male) recruited from the Queensland Bone Tumour Registry. Results showed prolonged activation of rectus femoris and prolonged co-contraction of the rectus femoris and hamstring muscles (p>0.05) in the affected limb of the limb salvage group compared to a control group. Prolonged rectus femoris activation and co-contraction was also evident in the unaffected lower limb suggesting alterations in gait programming within higher neuronal centres. The results are important for the development of rehabilitation programs as they suggest an overall reprogramming of the gait pattern, thereby limiting the impact of conventional strength and stretching interventions. PMID- 19560373 TI - Percutaneous removal of a metallic foreign body from the wall of the thoracic aorta. AB - This report details the percutaneous removal of a metallic foreign body from the wall of the thoracic aorta. The foreign body was presumably swallowed and migrated from the esophagus into the aortic lumen progressively during a period of 10 months. The patient had no adverse consequences from the procedure immediately or in the subsequent 18 months of follow-up. PMID- 19560374 TI - Percutaneous hepatic abscess drainage: do multiple abscesses or multiloculated abscesses preclude drainage or affect outcome? AB - PURPOSE: To compare the effectiveness of percutaneous abscess drainage in patients with pyogenic liver abscesses of the following types: single, single multiloculated, multiple, and multiple multiloculated. MATERIALS AND METHODS: One hundred nine patients with 149 liver abscesses who underwent percutaneous drainage during an 11-year period were divided into a single abscess group and a multiple abscess group. Of the 109 patients, 54 had multiloculated abscesses and were divided into single and multiple multiloculated abscess groups. Technical success was defined as the ability to place the catheter within the abscess cavity and clinical success was defined as improvement in the patient's symptoms. Clinical findings, management strategy, complication rate, and success rate were analyzed. RESULTS: Technical success rates were 96% (82 of 85) for a single abscess and 96% (23 of 24) for multiple abscesses (P = 1.0). Clinical success was achieved in 74 of 85 patients (87%) with a single abscess and 22 of 24 patients (92%) with multiple abscesses (P = .729). Technical success rates were 94% (32 of 34) for a single multiloculated abscess and 95% (19 of 20) for multiple multiloculated abscesses (P = 1.0). Clinical success was achieved in 30 of 34 patients (88%) with a single multiloculated abscess and 18 of 20 patients (90%) with multiple multiloculated abscesses (P = 1). No significant difference in hospital stay was seen between single and multiple abscess groups (P = .373) or between single multiloculated and multiple multiloculated abscess groups (P = .180). There were no major complications or mortality related to the procedure. CONCLUSIONS: Percutaneous drainage is a safe and effective procedure in the treatment of pyogenic liver abscess, regardless of abscess complexity and/or multiplicity. PMID- 19560375 TI - Long-term survival after radiofrequency ablation for pulmonary metastasis from hepatocellular carcinoma: report of two cases. PMID- 19560376 TI - Screening for peripheral arterial disease. PMID- 19560377 TI - Biodistribution of canine leucocytes labelled with technetium-99m stannous fluoride colloid in whole blood and their ability to localise to sites of induced inflammation. AB - This study assessed the biodistribution of autologous leucocytes radiolabelled with technetium-99m stannous fluoride colloid (99mTcSnC) for detection of foci of induced inflammation in dogs. Venous blood was collected from seven healthy dogs and incubated with 99mTcSnC for 1h at room temperature. Radiolabelled samples were injected intravenously (IV) and the dogs were scanned using a gamma camera. Another seven healthy dogs were injected intradermally with tumour necrosis factor alpha and then IV with 99mTcSnC radiolabelled autologous blood 3h later before being scanned. The radiolabelled leucocytes localised to sites of inflammation by 30 min post-injection. IV injection of autologous leucocytes radiolabelled with 99mTcSnC appears to be a sensitive method for localisation of induced foci of inflammation in dogs. PMID- 19560378 TI - Effect of chronic intra-peritoneally administered chimeric peptide of met enkephalin and FMRFa-[D-Ala2]YFa-on antinociception and opioid receptor regulation. AB - The physiological role of NPFF/FMRFa family of peptides is complex and exact mechanism of action of these peptides is not yet completely understood. In same line of scrutiny, previously we reported an enzymatically stable chimeric analog of YGGFMKKKFMRFamide (YFa) i.e., [D-Ala(2)]YAGFMKKKFMRFamide ([D-Ala(2)]YFa) which have a role in antinociception and modulatory effect on opioid analgesia. In continuation, presently we investigated using tail-flick test whether [D Ala(2)]YFa on systemic administration induced any antinociception in rats and if so then which specific opioid receptor(s) mu, delta or kappa mediated it. Further, the antinociceptive effect of [D-Ala(2)]YFa on 6 days chronic intra peritoneal (i.p.) treatment in rats was examined and finally, effect of this chronic treatment on the differential expression of opioid receptors was assessed. [D-Ala(2)]YFa on i.p. administration induced dose dependent antinociception which was mainly mediated by delta (DOR) and partially by mu (MOR) and kappa (KOR) opioid receptors. Moreover, its antinociceptive effect remained comparable throughout the chronic treatment even during insufficient availability of DOR1. Importantly, during this treatment the mRNA expression of all three opioid receptors (MOR1, KOR1 and DOR1) was increased as assessed by real-time RTPCR though subsequent western blot analysis revealed a selective increase in the protein level of DOR1, only. Thus, pharmacological behavior of [d Ala(2)]YFa suggests that competency of an opioid agonist to bind with multiple opioid receptors may enhance its potency to induce tolerance free analgesia. PMID- 19560379 TI - Persistent deep mechanical hyperalgesia induced by repeated cold stress in rats. AB - Chronic muscle pain of the neck, shoulder and low back is quite common and often related to a stressed condition. In this study we tried to make a model of long lasting muscle mechanical hyperalgesia based on one type of stress, repeated cold stress (RCS) (Kita T, Hata T, Yoneda R, Okage T. Stress state caused by alternation of rhythm in environmental temperature, and the functional disorders in mice and rats. Folia Pharmacol Jpn 1975;71:195-210). We first validated a method of measuring the muscle mechanical nociceptive threshold through skin, with surface anesthesia of the skin covering the muscle. We found that a pressure test using a Randall-Selitto analgesiometer equipped with a larger probe (varphi 2.6 mm) can measure the deep mechanical withdrawal threshold even under the presence of cutaneous punctuate hyperalgesia. RCS was performed by changing the temperature from 22 degrees C to either 4 degrees C (RCS at 4 degrees C) or -3 degrees C (RCS at -3 degrees C) every 30 min, and then maintained at 4 degrees C/ 3 degrees C from 17:30 to 10:00 the next day. RCS at 4 degrees C for 5 days induced bilateral deep mechanical hyperalgesia lasting 2-3 weeks without cutaneous punctuate hyperalgesia. Deep mechanical hyperalgesia observed after RCS at -3 degrees C lasted longer ( approximately 6 weeks) and was severer than RCS at 4 degrees C. Bilateral cutaneous punctuate hyperalgesia was also observed with RCS at -3 degrees C. Intramuscular injection of lidocaine confirmed that the muscle was hyperalgesic. RCS might serve as a useful model for study of the mechanism of chronic muscle pain and its treatment. PMID- 19560380 TI - The integrity of the anterior pretectal nucleus and dorsolateral funiculus is necessary for electroacupuncture-induced analgesia in the rat tail-flick test. AB - Previous studies have indicated that the anterior pretectal nucleus (APtN) is implicated in pathways that descend through the dorsolateral funiculus (DLF) to modulate nociceptive inputs in the spinal dorsal horn. The activation of descending inhibitory mechanisms also seems to be involved in electroacupuncture (EA)-induced analgesia. This study utilized the tail-flick test to examine the changes produced by DLF lesion or injection of 2% lidocaine into the APtN in the analgesia induced by 2 or 100 Hz EA applied to the Zusanli (ST36) and Sanyinjiao (SP6) acupoints in lightly anesthetized rats. Tail-flick latency was significantly increased by EA, the effect of 2 Hz EA lasting longer than that produced by 100 Hz EA. The effect of either 2 or 100 Hz EA did not occur in DLF lesion rats. The effect of 2 Hz EA did not occur in rats with neural block of the whole or dorsal APtN. In contrast, the effect of 100 Hz EA was reduced in rats with neural block of the whole APtN, but remained unchanged in rats with neural block of the dorsal APtN. We thus conclude that the integrity of the APtN and DLF is necessary for EA-induced analgesia in the rat tail-flick test. In addition, the integrity of the dorsal APtN is necessary for the analgesic effect of 2 but not 100 Hz EA. PMID- 19560382 TI - In vivo regulation of phenylalanine hydroxylase in the genetic mutant hph-1 mouse model. AB - The hph-1 mouse has low liver activity of GTP cyclohydrolase 1, the rate limiting enzyme in the biosynthesis of tetrahydrobiopterin (BH(4)). BH(4) is the cofactor for phenylalanine hydroxylase (PAH) and in the early stages of life the hph-1 mouse is hyperphenylalaninemic. At approximately 15 days after birth the blood phenylalanine levels normalize. During this period the animals provide an in vivo model which can be used to study the regulatory effects of phenylalanine on PAH, and for related pediatric metabolic disease in humans; from birth to youth. We therefore, examined; liver PAH activity using BH(4) and 6-methyltetrahydropterin (6MPH(4)) as cofactor; PAH total enzyme concentration by Western blotting using the PH8 antibody, and PAH state of phosphorylation using the PH7 antibody from 4 to 18 days after birth. The findings were compared to the wild type animals that are not hyperphenylalaninemic during this period. PAH (6MPH(4)) activity and total protein (PH8 antibody) rose steadily in the hph-1 mice. In control mice, both activity and total protein fluctuated. The degree of phosphorylation of PAH in the mutants and the state of activation (as measured by the 6MPH(4)/BH(4) activity ratio) increased as phenylalanine levels rose, and decreased when they fell. Similar patterns were not seen in the control animals. These studies provide in vivo evidence that phenylalanine concentration regulates the activity of PAH in the hph-1 mouse and that this acts via a mechanism that includes phosphorylation of the PAH molecule. The kinetic values (K(m) and V(max)) for mouse PAH are also reported. PMID- 19560381 TI - Evaluation of IGF-2/ApaI polymorphism in pregnant women infected with human immunodeficiency virus type 1 taking antiretroviral drugs. AB - OBJECTIVE: Studies carried out to assess the effects of antiretroviral drugs (ARV) in HIV-1 infected pregnant women have demonstrated carbohydrate intolerance. Some reports also refer to the effect of disturbances in the expression of the insulin-like growth factor (IGF) system on pancreas beta-cell function in humans and IGF-2/ApaI polymorphisms have been associated with obesity and features of the metabolic syndromes. In the present study, we tested the association between IGF-2/ApaI genotype and hyperglycemia in HIV-1 infected pregnant women receiving ARV. DESIGN: We studied IGF-2/ApaI polymorphism in 87 healthy pregnant women, 43 HIV-1 infected pregnant women taking ARV with hyperglycemia during pregnancy, and 43 HIV-1-negative pregnant women with gestational diabetes. Blood samples were obtained for DNA extraction, PCR and genotyping. Data were analyzed statistically by the Kolmogorov-Smirnov normality, ANOVA and chi-square tests. RESULTS: There were no significant differences in genotype frequency among the three groups analyzed. Considering the HIV-1 infected pregnant women, there were no significant differences in genotype frequency between the zidovudine group and the triple antiretroviral treatment group. There were no significant differences in allele frequencies among the groups evaluated. Non-white pregnant women tended to present the GG genotypes compared to white pregnant women. CONCLUSION: These results contribute to a better understanding of metabolic glycemic disorders in HIV-1-infected pregnant women using ARV, showing that IGF-2/ApaI polymorphisms are not responsible as a single causative factor of glycemic alterations. These data indicate that other variables should be studied in order to explain these glycemic abnormalities. PMID- 19560383 TI - HER-2/neu (c-erbB-2) oncoprotein in hyperplastic endometrial polyps detected in two cats. AB - The presence of HER-2/neu (c-erbB-2) oncoprotein, oestrogen-alpha receptor (ER), and progesterone receptor (PR) in hyperplastic endometrial polyps (EPs) of two cats with cystic endometrial hyperplasia-pyometra (CEH-P) complex was investigated. Immunohistochemistry assay for ER, PR and c-erbB-2 oncoprotein in the glandular and stromal tissue of the EPs was performed. ER and c-erbB-2 immunoreactivity was observed in the glandular epithelium of the EPs whereas PR immunoreactivity was detected only in the stromal fibroblasts. The c-erbB-2 oncoprotein may play a role with the ER in the pathogenesis of the hyperplastic EPs, although the role of this oncoprotein in the pathogenesis of EPs has yet to be determined. PMID- 19560384 TI - Dermoid cysts presenting as enlarged thyroid glands in a cat. AB - A 5-year-old spayed female cat was evaluated for hyperthyroidism based on an elevated free thyroxine (T(4)) measurement and bilaterally enlarged symmetric subcutaneous masses in the area of the thyroid glands. Physical examination revealed bilateral subcutaneous masses on either side of the cervical trachea. Blood was obtained for serum biochemical profile and thyroid function analysis. Mild hyperalbuminemia, mild hypercalcemia, and mildly increased alanine aminotransferase activity were identified. Serum concentrations of total and free thyroxine were within the reference interval. Cytologic analysis of fine-needle aspirates from one of the masses was suspicious for neoplasia. Nuclear scintigraphy revealed no abnormalities. Surgically obtained excisional biopsies of both masses were submitted for histopathology and diagnosed as bilateral dermoid cysts. After excisional biopsy, the patient recovered without incident. The histopathologic diagnosis of completely excised bilateral dermoid cysts indicated that no further medical or surgical intervention was required. This is the first report of a cat presenting with bilateral dermoid cysts in the area of the thyroid glands. Histopathologic examination was necessary to make a definitive diagnosis. Practitioners should include cysts in their list of differential diagnoses for ventral neck masses in cats. PMID- 19560386 TI - Two cases of feline ectromelia: autopodium ectromelia associated with humero ulnar synostosis and zeugopodium ectromelia. AB - Congenital limb deformities are rarely reported in cats. This paper describes the radiographic findings of congenital forelimb malformations in two cats. The radiographic changes were suggestive of an autopodium ectromelia associated with humero-ulnar synostosis in one case and zeugopodium ectromelia in the other case. Congenital feline limb deformities are poorly documented and, to the authors' knowledge, this is the first time that humero-ulnar synostosis has been reported in cats. PMID- 19560385 TI - Serum feline pancreatic lipase immunoreactivity concentration and seroprevalences of antibodies against Toxoplasma gondii and Bartonella species in client-owned cats. AB - Feline pancreatitis is a commonly suspected illness and it has been proposed that some cases of feline pancreatitis may be caused by infection with Toxoplasma gondii or Bartonella species. Feline pancreatic lipase immunoreactivity (fPLI) is a test performed on serum that is commonly combined with other clinical findings as an indirect aid in the diagnosis of pancreatitis. The purpose of this study was to determine if there are associations between fPLI concentration and the presence of serum antibodies against T gondii or Bartonella species. Serum samples from 458 cats, for which serum fPLI concentrations had already been determined, were assayed by enzyme-linked immunosorbent assay for the presence of T gondii immunoglobulin (Ig) G (IgG) and IgM antibodies, and Bartonella species IgG antibodies. The association between fPLI concentration and T gondii or Bartonella species antibodies was determined. No statistically significant association was found between fPLI concentration and T gondii or Bartonella species antibodies, suggesting that serological tests for the organisms are not useful in cases with increased fPLI concentration. PMID- 19560387 TI - Prevalence of feline coronavirus antibodies in cats in Bursa province, Turkey, by an enzyme-linked immunosorbent assay. AB - Feline sera from Bursa province (Turkey) were assayed for coronavirus antibody using an enzyme-linked immunosorbent assay (ELISA). The study was performed on 100 sera collected from cats belonging to catteries or community shelters and to households. The serum samples were initially tested with the virus neutralisation (VN) test and the results were then compared with the ELISA. The VN yielded 79 negative and 21 positive sera but the ELISA confirmed only 74 as negative. The ELISA-negative sera were also found to be free of feline coronoviruses-specific antibodies by Western blotting. Using the VN as the gold standard test, ELISA had a sensitivity of 100% and a specificity of 93.6%, with an overall agreement of 95%. The Kappa (kappa) test indicated high association between the two tests (kappa=0.86, 95% confidence interval (CI) 0.743-0.980). The positive predictive value (PPV) was 0.8, and the negative predictive value (NPV) was 0.93. The prevalence of FCoV II antibodies in the sampled population based on the gold standard was 62% (95% CI 0.44-0.77) among multi-cat environments, and 4% (95% CI 0.01-0.11) among single cat households. PMID- 19560388 TI - Multicentre, randomised, controlled study of the impact of continuous sub cutaneous glucose monitoring (GlucoDay) on glycaemic control in type 1 and type 2 diabetes patients. AB - AIM: This randomised study was designed to investigate the impact of continuous glucose monitoring (CGM) for 48h on glycaemic control with a 3-month follow-up in patients with type 1 (T1D) or type 2 (T2D) diabetes. METHODS: A total of 48 patients with poor glycaemic control (HbA(1c): 8-10.5%) underwent CGM for 48h using the GlucoDay((R)) system (A. Menarini Diagnostics), after which they were randomly assigned to treatment adjustments based on either their CGM profile (CGM group) or their usual self-monitoring of blood glucose (SMBG group). HbA(1c) measurement and 48-h CGM were repeated 3 months later. RESULTS: Altogether, 34 patients with either T1D (n=9) or T2D (n=25) completed the study; seven patients chose to leave the study, and seven patients in the CGM group were excluded because their baseline CGM graphs were not interpretable. HbA(1c) levels decreased significantly in the CGM group (n=14, -0.63+/-0.27%; P=0.023), but not in the controls (n=20, -0.28+/-0.21%; P=0.30). In T2D patients, the improvement associated with CGM vs SMBG was due to HbA(1c) decreases (mean: -0.63+/-0.34%; P=0.05 vs -0.31+/-0.29%; P=0.18, respectively). However, HbA(1c) did not change significantly with CGM in T1D patients. Comparisons of CGM data at baseline and after 3 months showed no significant changes in glucose control, glucose variability or hypoglycaemia. No major adverse events related to the GlucoDay system were reported. CONCLUSION: This is the first randomised study showing that CGM improves glycaemic control in patients with T2D. PMID- 19560390 TI - Vasculitis working group: selected unanswered questions related to giant cell arteritis and anti-neutrophil cytoplasmic antibody-associated vasculitis. AB - Evidence to guide assessment and management of patients with vasculitis is lacking for many important clinical questions. The evidence surrounding several common questions about management of vasculitis was reviewed. Patients with giant cell arteritis (GCA) are at risk for developing extra-cranial large vessel inflammation. Clinicians should be aware of this complication and search for large vessel involvement in patients with GCA who have ischemic symptoms. Research is needed to define optimal strategies to identify patients with such complications. Because of the hazards of chronic corticosteroid use, alternative therapies for patients with GCA have been sought but thus far no clear alternatives have been identified. Anti-neutrophil cytoplasmic antibodies (ANCA) are associated with small-vessel vasculitis, including Wegener's granulomatosis and microscopic polyangiitis, but changes in ANCA titers should not be used as a surrogate biomarker for disease activity. Several immunosuppressive agents can be used for maintenance therapy after induction of remission in patients with ANCA associated vasculitis, with no firm evidence that one agent is superior to others. Collectively, this review shows that more research is needed to provide a firmer body of evidence to support clinical decision-making for patients with vasculitis. PMID- 19560389 TI - Building muscle. PMID- 19560391 TI - Ultrasound-assisted third-liquid phase-transfer catalyzed esterification of sodium salicylate in a continuous two-phase-flow reactor. AB - The esterification of sodium salicylate to synthesize butyl salicylate by third liquid phase-transfer catalysis under ultrasound irradiation was investigated in a continuous two-phase-flow reactor. The reactor was designed to keep the third liquid phase in the middle part and to have the aqueous and organic phases flowing through it in countercurrent. Using tetra-n-butylphosphonium bromide to prepare the third-liquid phase for this esterification, the product yield in the organic outlet (toluene) at 70 degrees C was 49.7% in silent condition, showing the reaction promoted simply by countercurrent mixing of the aqueous and organic phases. In the conditions of space time at 168 min, stirring at 150 rpm and ultrasound irradiation (28 kHz, 300 W), the product yield was greatly enhanced to 78.2%. As prepared, above 90% of the added catalyst existed in the third-liquid phase, and after 4-h on stream for a large excess of n-butyl bromide to sodium salicylate, the fraction of catalyst retaining in the reactor was reduced to around 80%. The distributions of catalysts between phases before and after reaction were analyzed. A kinetic model was proposed to estimate the apparent rate constants, and the feasibility for third-liquid phase-transfer catalysis assisted by ultrasound irradiation in a continuous flow reactor was demonstrated. PMID- 19560392 TI - Impaired leg extensor strength in individuals with Parkinson disease and relatedness to functional mobility. AB - Previous investigations have demonstrated leg strength deficits in persons with Parkinson's disease (PD) as compared to neurologically-normal adults. However, the exact mode of contraction by which strength is assessed may determine how closely such deficits are related to functional performance. The purpose of this study was to better understand the relationship of strength and functional mobility in persons with PD (n = 17, mean H&Y stage = 2.0) via comparison to a group of similar age healthy controls (n = 10) using a multi-joint isometric test of strength and various measures of functional mobility. Tests included isometric leg press maximum force relative to body mass, the Activities-specific Balance Confidence scale (ABC), postural sway under various unilateral stance and visual conditions, and the timed up and go (TUG). Relative force (p = 0.044) and ABC questionnaire mean scores (p < 0.001), showed controls performing better than PD subjects. The control group performed better than the PD group for length of path of the center of pressure except in the eyes closed positions (p < 0.05 for all). TUG time (p = 0.052) was not significantly different between the PD group and healthy controls. Leg press maximum force relative to body mass was however significantly correlated with TUG time (r = -0.68, p = 0.003) in persons with PD. There were no gender differences for any variables. These results suggest that some balance and functional mobility task performances are more worse for persons with mild-to-moderate PD than for neurologically-normal age-matched controls, which may be influenced by lessened lower extremity multi-joint strength. Strength training of the lower extremity utilizing such multi-joint actions may be beneficial for this population. PMID- 19560393 TI - Brain-skull contact boundary conditions in an inverse computational deformation model. AB - Biomechanical models simulating brain motion under loading and boundary conditions in the operating room (OR) are gaining attention as alternatives for brain shift compensation during open cranial neurosurgeries. Although the significance of brain-skull boundary conditions (BCs) in these models has been explored in dynamic simulations, it has not been fully investigated in models representing the quasi-static brain motion that prevails during neurosurgery. In this study, we extend the application of a brain-skull contact BC by incorporating it into an inversion estimation scheme for the deformation field using the steepest gradient descent (SGD) framework. The technique allows parenchymal surface motion normal to the skull while maintaining stress-free BCs at the craniotomy and minimizing the effect of measurement noise. Application of the algorithm in five clinical cases using sparse data generated at the tumor boundary confirms the significance of brain-skull BCs in the model response. Specifically, the results demonstrate that the contact BC enhances model flexibility and achieves improved or comparable performance at the tumor boundary (recovering about 85% of the deformation) relative to that obtained when normal motion of the parenchymal surface is not allowed. It also significantly improves model estimation accuracy at the craniotomy (1.6mm on average), especially when the normal motion is large. The importance of the method is that model performance significantly improves when brain-skull contact influences the deformation field but does not degrade when the contact is less critical and simpler BCs would suffice. The computational cost of the technique is currently 3.9 min on average, but may be further reduced by applying an iterative solver to the linear systems of equations involved and/or by local refinement of the mesh in regions of interest. PMID- 19560394 TI - Recent advances in carbohydrate-based vaccines. AB - Vaccinations provide an efficient and cost-effective way to combat devastating human diseases. Besides pathogenic protein markers, cell surface carbohydrates from biological sources are widely used as vaccines. Recently, synthetic immunogenic carbohydrate-protein conjugates have been advanced to vaccine candidates. Progress in the chemical synthesis of oligosaccharides and conjugation methods stimulated the development of novel carbohydrate-based vaccine candidates. PMID- 19560395 TI - Synthesis and characterization of toluene-3,4-dithiolatoantimony(III) derivatives with some oxygen and/or sulphur donor ligands. AB - Replacement reactions of toluene-3,4-dithiolatoantimony(III) chloride with oxygen and/or sulphur donor ligands like benzoic acid, thiobenzoic acid, thioacetic acid, phenol, thiophenol, sodium salicylate and thio glycolic acid in 1:1 molar ratio as well as disodium oxalate in 2:1 molar ratio in refluxing anhydrous benzene yielded toluene-3,4-dithiolatoantimony(III) mono oxo and/or thio carboxylic or phenolic derivatives of the general formula {R=OOCC(6)H(5), SOCC(6)H(5), SOCCH(3), OC(6)H(5), SC(6)H(5), OOCC(6)H(4)(OH) and SCH(2)COOH} and These newly synthesized derivatives are yellow and brown solids/liquids and are soluble in common organic solvents like benzene, chloroform, dichloromethane, etc. These derivatives have been characterized by melting point determination, molecular weight determination, elemental analysis (C, H, S and Sb), spectral {UV, IR and NMR ((1)H and (13)C)} and thermal (TGA, DTA and DSC) studies. PMID- 19560396 TI - Triplet behavior in weakly coupled chromophors in covalent pyridyl porphyrin polymer systems. AB - The photophysical properties of the 5-(4-pyridyl)-10,15,20-tri(4 methyloxyphenyl)porphyrins covalently linked to polyethylene glycol - PEG of different molecular weights (35000, 20000 and 8000) dissolved in dimethylsulfoxide were studied. The singlet and triplet states of the porphyrin species behavior were discussed in terms of fluorescence and thermal relaxation processes. The absorption, fluorescence and photothermal experiments showed that in the porphyrins linked to the PEG systems in dimethylsulfoxide the dye moieties occur in weakly interacting dimers. The triplet state enhancement in the 5-(4 pyridyl)-10,15,20-tri(4-methyloxyphenyl)porphyrins covalently linked to PEG was discussed. It was shown that even that the weak interaction of the porphyrin species in the covalent systems with PEG is not detectable by the absorption and only slightly by fluorescence, it is possible to be performed by the complementary spectroscopic methods like photoacoustics and photothermal time resolved spectroscopy. PMID- 19560397 TI - Hepatitis B virus reactivation in a large haemodialysis unit: virological and infection control issues. PMID- 19560398 TI - Measurements of HCV neutralizing antibodies and of HCV-specific CD4+ and CD8+ cells using hepatitis C virus pseudo-particles (HCVpp). AB - BACKGROUND: For the medical management, it would be of great relevance to get a diagnostic marker predicting the outcome of infection. OBJECTIVES: For this purpose, the envelope antigens of the individual HCV strain in a patient was tested for their capacity to induce neutralizing antibodies and cytotoxic T lymphocytes. STUDY DESIGN: A system for the measurement of neutralizing antibodies as well as for the stimulation of a HCV-specific T-cell response using pseudo-typed HCV particles (HCVpp) was established. A report on results of a pilot study conducted with blood specimens of 19 chronically infected patients is also presented. RESULTS: Neutralization of HCVpp could be measured in nearly all HCV sero-positive patient samples. Nevertheless, in more than half of the patient samples (11/19), no HCV-specific CD4+ response was detectable. In addition, HCV specific CD8+ response was measurable in most of the patients when HCVpp were used for T-cell stimulation. Although the same antigens (HCVpp) were used, there was no relevant correlation between neutralization titers and T-cell response. CONCLUSION: Our study shows that HCVpp are appropriate antigens for specific stimulation of lymphocytes as well as for the investigation of antibody neutralization activity. PMID- 19560399 TI - Safety of 1 Hz repetitive transcranial magnetic stimulation (rTMS) in patients with titanium skull plates. PMID- 19560400 TI - Maturation of somatosensory cortical processing from birth to adulthood revealed by magnetoencephalography. AB - OBJECTIVE: To evaluate the maturation of tactile processing by recording somatosensory evoked magnetic fields (SEFs) from healthy human subjects. METHODS: SEFs to tactile stimulation of the left index finger were measured from the contralateral somatosensory cortex with magnetoencephalography (MEG) in five age groups: newborns, 6- and 12-18-month-olds, 1.6-6-year-olds, and adults. The waveforms of the measured signals and equivalent current dipoles (ECDs) were analyzed in awake and sleep states in order to separate the effects of age and vigilance state on SEFs. RESULTS: There was an orderly, systematic change in the measured and ECD source waveforms of the initial cortical responses with age. The broad U-shaped response in newborns (M60) shifted to a W-shaped response with emergence of a notch by 6 months of age. The adult-type response with M30 and M50 components was present by 2 years. The ECDs of M60 and M30 were oriented anteriorly and that of M50 posteriorly. These maturational changes were independent of vigilance state. CONCLUSIONS: The most significant maturation of short latency cortical responses to tactile stimulation takes place during the first 2 years of life. SIGNIFICANCE: The maturational changes of somatosensory processing can noninvasively be evaluated with MEG already in infancy. PMID- 19560401 TI - Annotation an effective device for student feedback: a critical review of the literature. AB - The paper examines hand-written annotation, its many features, difficulties and strengths as a feedback tool. It extends and clarifies what modest evidence is in the public domain and offers an evaluation of how to use annotation effectively in the support of student feedback [Marshall, C.M., 1998a. The Future of Annotation in a Digital (paper) World. Presented at the 35th Annual GLSLIS Clinic: Successes and Failures of Digital Libraries, June 20-24, University of Illinois at Urbana-Champaign, March 24, pp. 1-20; Marshall, C.M., 1998b. Toward an ecology of hypertext annotation. Hypertext. In: Proceedings of the Ninth ACM Conference on Hypertext and Hypermedia, June 20-24, Pittsburgh Pennsylvania, US, pp. 40-49; Wolfe, J.L., Nuewirth, C.M., 2001. From the margins to the centre: the future of annotation. Journal of Business and Technical Communication, 15(3), 333 371; Diyanni, R., 2002. One Hundred Great Essays. Addison-Wesley, New York; Wolfe, J.L., 2002. Marginal pedagogy: how annotated texts affect writing-from source texts. Written Communication, 19(2), 297-333; Liu, K., 2006. Annotation as an index to critical writing. Urban Education, 41, 192-207; Feito, A., Donahue, P., 2008. Minding the gap annotation as preparation for discussion. Arts and Humanities in Higher Education, 7(3), 295-307; Ball, E., 2009. A participatory action research study on handwritten annotation feedback and its impact on staff and students. Systemic Practice and Action Research, 22(2), 111-124; Ball, E., Franks, H., McGrath, M., Leigh, J., 2009. Annotation is a valuable tool to enhance learning and assessment in student essays. Nurse Education Today, 29(3), 284-291]. Although a significant number of studies examine annotation, this is largely related to on-line tools and computer mediated communication and not hand written annotation as comment, phrase or sign written on the student essay to provide critique. Little systematic research has been conducted to consider how this latter form of annotation influences student learning and assessment or, indeed, helps tutors to employ better annotative practices [Juwah, C., Macfarlane Dick, D., Matthew, B., Nicol, D., Ross, D., Smith, B., 2004. Enhancing student learning through effective formative feedback. The Higher Education Academy, 1 40; Jewitt, C., Kress, G., 2005. English in classrooms: only write down what you need to know: annotation for what? English in Education, 39(1), 5-18]. There is little evidence on ways to heighten students' self-awareness when their essays are returned with annotated feedback [Storch, N., Tapper, J., 1997. Student annotations: what NNS and NS university students say about their own writing. Journal of Second Language Writing, 6(3), 245-265]. The literature review clarifies forms of annotation as feedback practice and offers a summary of the challenges and usefulness of annotation. PMID- 19560402 TI - Urethral ratio on voiding cystourethrogram: a comparative method to assess success of posterior urethral valve ablation. AB - OBJECTIVE: To develop a simple, objective and reproducible quantitative measurement to assess success of posterior urethral valve ablation. METHOD: In 30 patients with posterior urethral valves the diagnosis was confirmed by voiding cystourethrogram (VCUG). Our protocol was to perform valve ablation, and repeat VCUG at 12 weeks postoperatively. Urethral ratio was calculated by dividing the posterior urethral diameter by the anterior urethral diameter. Thirty males undergoing VCUG for urinary tract infections were evaluated as normative controls. RESULTS: Median age of controls was 12 months (2 days-6 years) and of study group was 13 months (1 day-11 years). Mean urethral ratio in pre fulguration group was 4.94 (+/-2.97) and in post-fulguration group was 2.134 (+/ 1.19) (P<0.001). The mean urethral ratio in the control group of 1.73 (+/-0.577) was significantly different from the pre-fulguration group result (P<0.001), but not significantly different in comparison to the post-fulguration group (P=0.104). CONCLUSION: Calculation of urethral ratio on VCUG as a method of assessment of outcome of fulguration is objective, reproducible, and allows preoperative and postoperative VCUG from different facilities to be compared. A post-fulguration urethral ratio of 2.5-3 represents an acceptable result postoperatively. PMID- 19560404 TI - Point of view: spinopelvic parameters in postfusion flatback deformity patients. PMID- 19560405 TI - Kv7.1 in atrial fibrillation. PMID- 19560403 TI - Neuroanatomical correlates of cognitive phenotypes in temporal lobe epilepsy. AB - OBJECTIVE: Previous research characterized three cognitive phenotypes in temporal lobe epilepsy, each associated with a different profile of clinical seizure and demographic characteristics, total cerebral (gray, white, cerebrospinal fluid) and hippocampal volumes, and prospective cognitive trajectories. The objective of this investigation was to characterize in detail the specific neuroanatomical abnormalities associated with each cognitive phenotype. METHODS: High-resolution MRI scans of healthy controls (n=53) and patients with temporal lobe epilepsy (n=55), grouped by cognitive phenotype (minimally impaired; memory impaired; memory, executive function, and speed impaired), were examined with respect to patterns of gray matter thickness throughout the cortical mantle, as well as volumes of subcortical structures, corpus callosum, and regions of the cerebellum. RESULTS: Increasing abnormalities in temporal and extratemporal cortical thickness, volumes of subcortical structures (hippocampus, thalamus, basal ganglia), all regions of the corpus callosum, and bilateral cerebellar gray matter distinguish the cognitive phenotypes in a generally stepwise fashion. The most intact anatomy is observed in the minimally impaired epilepsy group and the most abnormal anatomy is evident in the epilepsy group with impairments in memory, executive function, and speed. CONCLUSION: Empirically derived cognitive phenotypes are associated with the presence, severity, and distribution of anatomic abnormalities in widely distributed cortical, subcortical, callosal, and cerebellar networks. PMID- 19560406 TI - Compound heterozygous SCN5A mutations: does the sum of the parts equal the whole? PMID- 19560407 TI - Comment on: Patterns of readmission and reoperation within 90 days after Roux-en Y gastric bypass. PMID- 19560408 TI - Development and validation of an improved liquid chromatography-mass spectrometry method for the determination of pemetrexed in human plasma. AB - An improved liquid chromatography-mass spectrometry method for the determination of pemetrexed in human plasma was developed and validated using a simple quadrupole LC-MS and a new SPE cartridge (Plexa Bond Elut). The analysis was achieved with a C18 analytical column using a mobile phase consisting of formic acid/acetonitrile and isocratic flow for 7 min. The linear ranges (r(2)>0.99) were found from 5 to 5000 ng/mL. The lower limit of detection was 2.5 ng/mL. Within-day and between-day precisions were less than 7.2% and inaccuracy did not exceed 2.8%. This new method is suitable to support pharmacokinetic studies and drug monitoring. PMID- 19560409 TI - Production and characterization of a genetically engineered anti-caffeine camelid antibody and its use in immunoaffinity chromatography. AB - This work demonstrates the feasibility of using a camelid single domain antibody for immunoaffinity chromatographic separation of small molecules. An anti caffeine VHH antibody was produced by grafting the complementarity determining sequences of a previously generated antibody onto an anti-RNase A antibody scaffold, followed by expression in E. coli. Analysis of the binding properties of the antibody by ELISA and fluorescence-based thermal shift assays showed that it recognizes not only caffeine, but also theophylline, theobromine, and paraxanthine, albeit with lower affinity. Further investigation of the effect of environmental conditions, i.e., temperature, pH, and ionic strength, on the antibody using these methods provided useful information about potential elution conditions to be used in chromatographic applications. Immobilization of the VHH onto a high flow-through synthetic support material resulted in a stationary phase capable of separating caffeine and its metabolites. PMID- 19560410 TI - Development and validation of a capillary zone electrophoresis method for assessment of recombinant human granulocyte colony-stimulating factor in pharmaceutical formulations and its correlation with liquid chromatography methods and bioassay. AB - A capillary zone electrophoresis (CZE) method was validated for the analysis of recombinant human granulocyte colony-stimulating factor (rhG-CSF) and performed on a fused-silica capillary, with detection at 195 nm. The background electrolyte solution consisted of 50 mM sodium tetraborate solution at pH 9. The method was linear in the concentration range of 1-200 microg/mL and the limit of quantitation (LOQ) was 1 microg/mL, with acceptable validation parameters. The method was applied for the analysis of pharmaceutical formulations, and the results were correlated to the reversed-phase HPLC method (RP-HPLC), size exclusion HPLC method (SE-HPLC) and in vitro bioassay method. PMID- 19560411 TI - Casting weight goal as a function of weight status among a representative population-based sample of adolescents. AB - This paper pursues two objectives: (1) to estimate proportions of adolescents with a weight goal that matched and mismatched weight status, and (2) to identify correlates of a mismatched weight goal. Data were from a representative population-based sample of adolescents (n=2346, 51% female; 91.5% complete data). Results showed that 69% of adolescents had a weight goal that matched weight status whereas 31% had a weight goal that mismatched weight status. Body dissatisfaction was a significant predictor of having a mismatched weight goal for both sexes while elevated psychological distress was a predictor among girls. Being body dissatisfied mediated the association between psychological distress and having a mismatched weight goal among girls. Casting weight goal as a function of weight status may allow for a better understanding of overall weight management strategies. PMID- 19560412 TI - Experience with the Mentor Contour Profile Becker-35 expandable implants in reconstructive breast surgery. AB - INTRODUCTION: Round expander-implants (Beckers 25 and 50) and anatomical expander prostheses filled with firm cohesive gel (McGhan Style 150) are established choices for single-stage expander breast reconstruction. Because of their drawbacks we selectively adopted the anatomical Becker-35 expander-implant filled with soft cohesive gel from January 2005. PATIENTS AND METHODS: All patients undergoing reconstructive breast surgery using the Contour Profile Becker-35 expandable implant over a two-year period were retrospectively reviewed with respect to indication, implant sizes, inflation details, complications and outcomes. RESULTS: 36 patients, mean age 48.9 years (r=14-69), received 39 anatomical Becker-35 expanders (three bilaterally). Three quarters of these implants (29) were used for immediate breast reconstruction while the remainder were equally divided between delayed postmastectomy reconstruction (5) and correction of congenital breast asymmetry (5). Half of the patients had simultaneous latissimus dorsi myocutaneous flap coverage of the implants. The median numbers of inflations and deflations needed to achieve the target expansion size and shape were 3 (r=0-7) and 0 (r=0-4), respectively. The mean time from expander insertion to completion of reconstruction was 4.6 months (r=0 13 months). Four patients required surgical intervention for haematoma, implant infection, severe capsular contracture, and palpable rippling. Additionally there were three injection port adjustments, giving a 20% overall revisional surgery rate (8/39 breasts) after a median follow-up of 20 months (r=6-38 months). Four implants (10%) developed significant but asymptomatic rippling. The significant capsular contracture rate was 21% (8/39 breasts), which was related to chest wall radiotherapy. CONCLUSION: In this short-term study, the Becker-35 expander was successfully used for single-stage prosthetic breast reconstruction with an incidence of early complications comparable to alternative prostheses. Although it has expanded the range of implants available to the breast surgeon, its exact role in reconstructive breast surgery has yet to be established. PMID- 19560413 TI - [Indexing glomerular filtration rate for body surface area: myth and reality]. AB - Indexing glomerular filtration rate (GFR) for body surface area (BSA) is often realized without arising any questions. However, physiological basis for such an indexation are very poor. Indeed, indexing GFR for BSA to avoid variation due to differences in body size necessarily implies that GFR is a linear function of BSA and that the intercept of this linear function is zero. Moreover, when GFR is indexed for BSA, the relation indexed GFR-BSA must completely disappear. These physiological prerequisites are not found for BSA indexation. We will review the history of this indexation and will underline errors and defaults. Different equations to estimate BSA exist and will be discussed. The choice of "1.73m(2)" will be also criticized. Moreover, indexing GFR for BSA has little impact on GFR results in "normal" body size population. Nevertheless, this indexation will have strong consequences in very lean (such as anorexia) and in obese patients. We will discuss possible alternatives proposed to substitute for BSA indexation. PMID- 19560414 TI - The molecular dating game: an antibody heavy chain hangs loose with a chaperone while waiting for its life partner. AB - In a recent issue of Molecular Cell, Feige et al. (2009) utilize the murine immunoglobulin system to shed light on a long-standing puzzle: how do cells coordinate folding of different polypeptides that ultimately form a complex? PMID- 19560415 TI - The guardian recruits cops: the p53-p21 axis delegates prosurvival duties to the Keap1-Nrf2 stress pathway. AB - In this issue of Molecular Cell, Zhang and colleagues (Chen et al., 2009) report their identification of p21 as a positive regulator of Nrf2, which extends p53 prosurvival functions through recruitment of a master stress-response regulator. PMID- 19560416 TI - A histone code for regulating V(D)J recombination. AB - In a recent issue of Molecular Cell, Shimazaki et al. (2009) show that an interaction between RAG2 and a methylated histone might play a critical regulatory role in V(D)J recombination by enhancing DNA binding and enzymatic activity of the V(D)J recombinase. PMID- 19560417 TI - The juxtamembrane region of the EGF receptor functions as an activation domain. AB - In several growth factor receptors, the intracellular juxtamembrane (JM) region participates in autoinhibitory interactions that must be disrupted for tyrosine kinase activation. Using alanine scanning mutagenesis and crystallographic approaches, we define a domain within the JM region of the epidermal growth factor receptor (EGFR) that instead plays an activating--rather than autoinhibitory--role. Mutations in the C-terminal 19 residues of the EGFR JM region abolish EGFR activation. In a crystal structure of an asymmetric dimer of the tyrosine kinase domain, the JM region of an acceptor monomer makes extensive contacts with the C lobe of a donor monomer, thus stabilizing the dimer. We describe how an uncharacterized lung cancer mutation in this JM activation domain (V665M) constitutively activates EGFR by augmenting its capacity to act as an acceptor in the asymmetric dimer. This JM mutant promotes cellular transformation by EGFR in vitro and is tumorigenic in a xenograft assay. PMID- 19560418 TI - KSR2 is a calcineurin substrate that promotes ERK cascade activation in response to calcium signals. AB - Protein scaffolds have emerged as important regulators of MAPK cascades, facilitating kinase activation and providing crucial spatio/temporal control to their signaling outputs. Using a proteomics approach to compare the binding partners of the two mammalian KSR scaffolds, we find that both KSR1 and KSR2 interact with the kinase components of the ERK cascade and have a common function in promoting RTK-mediated ERK signaling. Strikingly, we find that the protein phosphatase calcineurin selectively interacts with KSR2 and that KSR2 uniquely contributes to Ca2+-mediated ERK signaling. Calcineurin dephosphorylates KSR2 on specific sites in response to Ca2+ signals, thus regulating KSR2 localization and activity. Moreover, we find that depletion of endogenous KSR2 impairs Ca2+ mediated ERK activation and ERK-dependent signaling responses in INS1 pancreatic beta-cells and NG108 neuroblastoma cells. These findings identify KSR2 as a Ca2+ regulated ERK scaffold and reveal a new mechanism whereby Ca2+ impacts Ras to ERK pathway signaling. PMID- 19560419 TI - Direct interaction between Nrf2 and p21(Cip1/WAF1) upregulates the Nrf2-mediated antioxidant response. AB - In response to oxidative stress, Nrf2 and p21(Cip1/WAF1) are both upregulated to protect cells from oxidative damage. Nrf2 is constantly ubiquitinated by a Keap1 dimer that interacts with a weak-binding (29)DLG motif and a strong-binding (79)ETGE motif in Nrf2, resulting in degradation of Nrf2. Modification of the redox-sensitive cysteine residues on Keap1 disrupts the Keap1-(29)DLG binding, leading to diminished Nrf2 ubiquitination and activation of the antioxidant response. However, the underlying mechanism by which p21 protects cells from oxidative damage remains unclear. Here we present molecular and genetic evidence suggesting that the antioxidant function of p21 is mediated through activation of Nrf2 by stabilizing the Nrf2 protein. The (154)KRR motif in p21 directly interacts with the (29)DLG and (79)ETGE motifs in Nrf2 and thus competes with Keap1 for Nrf2 binding, compromising ubiquitination of Nrf2. Furthermore, the physiological significance of our findings was demonstrated in vivo using p21 deficient mice. PMID- 19560420 TI - Allosteric activation of E2-RING finger-mediated ubiquitylation by a structurally defined specific E2-binding region of gp78. AB - The activity of RING finger ubiquitin ligases (E3) is dependent on their ability to facilitate transfer of ubiquitin from ubiquitin-conjugating enzymes (E2) to substrates. The G2BR domain within the E3 gp78 binds selectively and with high affinity to the E2 Ube2g2. Through structural and functional analyses, we determine that this occurs on a region of Ube2g2 distinct from binding sites for ubiquitin-activating enzyme (E1) and RING fingers. Binding to the G2BR results in conformational changes in Ube2g2 that affect ubiquitin loading. The Ube2g2:G2BR interaction also causes an approximately 50-fold increase in affinity between the E2 and RING finger. This results in markedly increased ubiquitylation by Ube2g2 and the gp78 RING finger. The significance of this G2BR effect is underscored by enhanced ubiquitylation observed when Ube2g2 is paired with other RING finger E3s. These findings uncover a mechanism whereby allosteric effects on an E2 enhance E2-RING finger interactions and, consequently, ubiquitylation. PMID- 19560421 TI - Glutamine-specific N-terminal amidase, a component of the N-end rule pathway. AB - Deamidation of N-terminal Gln by Nt(Q)-amidase, an N-terminal amidohydrolase, is a part of the N-end rule pathway of protein degradation. We detected the activity of Nt(Q)-amidase, termed Ntaq1, in mouse tissues, purified Ntaq1 from bovine brains, identified its gene, and began analyzing this enzyme. Ntaq1 is highly conserved among animals, plants, and some fungi, but its sequence is dissimilar to sequences of other amidases. An earlier mutant in the Drosophila Cg8253 gene that we show here to encode Nt(Q)-amidase has defective long-term memory. Other studies identified protein ligands of the uncharacterized human C8orf32 protein that we show here to be the Ntaq1 Nt(Q)-amidase. Remarkably, "high-throughput" studies have recently solved the crystal structure of C8orf32 (Ntaq1). Our site directed mutagenesis of Ntaq1 and its crystal structure indicate that the active site and catalytic mechanism of Nt(Q)-amidase are similar to those of transglutaminases. PMID- 19560422 TI - Cellular microRNA and P bodies modulate host-HIV-1 interactions. AB - MicroRNAs (miRNAs), approximately 22 nt noncoding RNAs, assemble into RNA-induced silencing complexes (RISCs) and localize to cytoplasmic substructures called P bodies. Dictated by base-pair complementarity between miRNA and a target mRNA, miRNAs specifically repress posttranscriptional expression of several mRNAs. Here we report that HIV-1 mRNA interacts with RISC proteins and that disrupting P body structures enhances viral production and infectivity. In HIV-1-infected human T lymphocytes, we identified a highly abundant miRNA, miR-29a, which specifically targets the HIV-1 3'UTR region. Inhibiting miR-29a enhanced HIV-1 viral production and infectivity, whereas expressing a miR-29 mimic suppressed viral replication. We also found that specific miR-29a-HIV-1 mRNA interactions enhance viral mRNA association with RISC and P body proteins. Thus we provide an example of a single host miRNA regulating HIV-1 production and infectivity. These studies highlight the significance of miRNAs and P bodies in modulating host cell interactions with HIV-1 and possibly other viruses. PMID- 19560423 TI - Structural basis of transcription: mismatch-specific fidelity mechanisms and paused RNA polymerase II with frayed RNA. AB - We show that RNA polymerase (Pol) II prevents erroneous transcription in vitro with different strategies that depend on the type of DNARNA base mismatch. Certain mismatches are efficiently formed but impair RNA extension. Other mismatches allow for RNA extension but are inefficiently formed and efficiently proofread by RNA cleavage. X-ray analysis reveals that a TU mismatch impairs RNA extension by forming a wobble base pair at the Pol II active center that dissociates the catalytic metal ion and misaligns the RNA 3' end. The mismatch can also stabilize a paused state of Pol II with a frayed RNA 3' nucleotide. The frayed nucleotide binds in the Pol II pore either parallel or perpendicular to the DNA-RNA hybrid axis (fraying sites I and II, respectively) and overlaps the nucleoside triphosphate (NTP) site, explaining how it halts transcription during proofreading, before backtracking and RNA cleavage. PMID- 19560424 TI - Highly transcribed RNA polymerase II genes are impediments to replication fork progression in Saccharomyces cerevisiae. AB - Replication forks face multiple obstacles that slow their progression. By two dimensional gel analysis, yeast forks pause at stable DNA protein complexes, and this pausing is greatly increased in the absence of the Rrm3 helicase. We used a genome-wide approach to identify 96 sites of very high DNA polymerase binding in wild-type cells. Most of these binding sites were not previously identified pause sites. Rather, the most highly represented genomic category among high DNA polymerase binding sites was the open reading frames (ORFs) of highly transcribed RNA polymerase II genes. Twice as many pause sites were identified in rrm3 compared with wild-type cells, as pausing in this strain occurred at both highly transcribed RNA polymerase II genes and the previously identified protein DNA complexes. ORFs of highly transcribed RNA polymerase II genes are a class of natural pause sites that are not exacerbated in rrm3 cells. PMID- 19560426 TI - The ordered transcription of RNA domains is not essential for ribosome biogenesis in Escherichia coli. AB - Ribosome biogenesis is coupled to the transcription of ribosomal (r)RNAs, which enables an ordered and hierarchical assembly of the ribosome starting with the 5' terminal domain. We constructed four circular permutants (CPs) of Escherichia coli rRNAs in which the original termini of 16S or 23S rRNAs were genetically connected, and new termini were created elsewhere within the same rRNAs (in helix 33 of 16S rRNA, or in helices 45, 63, and 78 of 23S rRNA). Unexpectedly, all CPs tested were able to rescue E. coli strain Delta7 prrn, which lacks all chromosomal rRNA operons. This result demonstrates that hierarchical assembly from the 5'-terminal domain of both 16S and 23S rRNAs is not essential for ribosomal formation in the cell. However, severe growth defects of all CPs were found in the absence of the DEAD box RNA helicase deaD, indicating that DeaD assists in the efficient assembly of each subunit in the cell. PMID- 19560425 TI - Mechanical constraints on Hin subunit rotation imposed by the Fis/enhancer system and DNA supercoiling during site-specific recombination. AB - Hin, a member of the serine family of site-specific recombinases, regulates gene expression by inverting a DNA segment. DNA inversion requires assembly of an invertasome complex in which a recombinational enhancer DNA segment bound by the Fis protein associates with the Hin synaptic complex at the base of a supercoiled DNA branch. Each of the four Hin subunits becomes covalently joined to the cleaved DNA ends, and DNA exchange occurs by translocation of a Hin subunit pair within the tetramer. We show here that, although the Hin tetramer forms a bidirectional molecular swivel, the Fis/enhancer system determines both the direction and number of subunit rotations. The chirality of supercoiling directs rotational direction, and the short DNA loop stabilized by Fis-Hin contacts limit rotational processivity, thereby ensuring that the DNA strands religate in the recombinant configuration. We identify multiple rotational conformers that are formed under different supercoiling and solution conditions. PMID- 19560427 TI - Shifts in replication timing actively affect histone acetylation during nucleosome reassembly. AB - The entire genome is replicated in a programmed manner, with specific regions undergoing DNA synthesis at different times in S phase. Active genes generally replicate in early S phase, while repressed genes replicate late, and for some loci this process is developmentally regulated. Using a nuclear microinjection system, we demonstrate that DNA sequences originally packaged into nucleosomes containing deacetylated histones during late S become reassembled with acetylated histones after undergoing replication in early S. Conversely, a change from early to late replication timing is accompanied by repackaging into nucleosomes containing deacetylated histones. This is carried out by differential cell-cycle controlled acetylation and deacetylation of histones H3 and H4. These studies provide strong evidence that switches in replication timing may play a role in the regulation of nucleosome structure during development. PMID- 19560429 TI - Apropos "vaccines in a hurry". PMID- 19560430 TI - The fight against rabies in Africa: From recognition to action. AB - As a follow-up to the first AfroREB meeting, held in Grand Bassam (Cote d'Ivoire) in March 2008, African rabies experts of the AfroREB network met a second time to complete the evaluation of the rabies situation in Africa and define specific action plans. About 40 French speaking rabies specialists from northern, western and central Africa and Madagascar met in Dakar (Senegal), from 16 to 19 March 2009. With the participation of delegates from Tunisia, who joined the AfroREB network this year, 15 French speaking African countries were represented. Experts from the Institut Pasteur in Paris, the Alliance for Rabies Control, and the Southern and Eastern African Rabies Group (SEARG, a network of rabies experts from 19 English speaking Southern and Eastern African countries) were in attendance, to participate in the discussion and share their experiences. It was unanimously agreed that the priority is to break the vicious cycle of indifference and lack of information which is the main barrier to human rabies prevention. PMID- 19560431 TI - Giardia intestinalis genotypes: Risk factors and correlation with clinical symptoms. AB - This study was conducted to identify genotypes related risk factors of Giardia intestinalis in an Orang Asli (aboriginal) community in Pahang, Malaysia. Stool samples were collected from 321 individuals aged between 2 and 76 years old, of whom 160 were males and 161 were females. Faecal samples were processed with trichrome staining technique for the primary identification of G. intestinalis. Molecular identification was carried out by the amplification of a partial SSU rRNA gene using nested PCR. PCR products were purified and genotyped. 42 samples successfully amplified from the 76 positive faecal samples, only 1 was Assemblage A, the rest were Assemblage B. Risk analysis based on the detected genotypes of Giardia using univariate analysis and logistic regression identified three significant risk factors of giardiasis caused by assemblage B which included children 0.05), but significant between ELISA and COPT assays (chi(2)=6.72, P<0.01). Our results also revealed significant difference in positive rate between ELISA and IHA (chi(2)=24.74, P<0.005), as well as between ELISA and COPT (chi(2)=58.14, P<0.005). These results suggest that the rSj14-3-3 and r26kDa SjGST would be effective diagnostic antigens for detection of antibodies to S. japonicum in human. Due to the easy production, high sensitivity and specificity, the recombinant proteins tested in this study can be considered as candidate reagent for immunological diagnosis of human schistosomiasis. PMID- 19560434 TI - Colorimetric microbial viability assay based on reduction of water-soluble tetrazolium salts for antimicrobial susceptibility testing and screening of antimicrobial substances. AB - The applicability of a colorimetric microbial viability assay based on reduction of a tetrazolium salt {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4 disulfophenyl)-2H-tetrazolium, monosodium salt [WST-8]} via 2-methyl-1,4 naphthoquinone (2-methyl-1,4-NQ) as an electron mediator for determining the susceptibility of various bacteria to antibiotics and screening antimicrobial substances was investigated. The measurement conditions, which include the effects of the concentration of 2-methyl-1,4-NQ, were optimized for proliferation assays of gram-negative bacteria, gram-positive bacteria, and pathogenic yeast. In antimicrobial susceptibility testing, there was excellent agreement between the minimum inhibitory concentrations determined after 8 h using the WST-8 colorimetric method and those obtained after 22 h using conventional methods. The results suggest that the WST-8 colorimetric assay is a useful method for rapid determination of the susceptibility of various bacteria to antibiotics. In addition, the current method was applied to the screening of bacteriocin producing lactic acid bacteria and its efficiency was demonstrated. PMID- 19560433 TI - A biotin-based protein tagging system. AB - Biotin protein ligase (BPL) mediates covalent attachment of biotin to a specific lysine residue of biotin carboxyl carrier protein (BCCP) of biotin-dependent enzymes. We recently found that the biotinylation reaction from thermophilic archaeon Sulfolobus tokodaii has a unique characteristic that the enzyme BPL forms a tight complex with the product, biotinylated BCCP (169 amino acid residues). In the current work, we attempted to apply this characteristic to a novel protein tagging system. Thus, the N terminus of S. tokodaii BCCP was truncated and the interaction of the resulting BCCP, BCCPDelta100 and BCCPDelta17 (with 69 and 152 residues, respectively), with BPL was investigated by surface plasmon resonance (SPR). It was found that the binding of BPL to the biotinylated BCCPDelta100 is extremely tight with a dissociation constant (K(D)) of 1.2 nM, whereas that to the unbiotinylated counterpart was moderate with a K(D) of 3.3 microM. Furthermore, chimeric proteins of glutathione S-transferase (GST) and green fluorescence protein (GFP) with BCCPDelta100 fused to their C terminus were prepared. The resulting fusion proteins were successfully biotinylated and captured on the BPL-modified SPR sensor chip or BPL-modified magnetic beads. The function of GST and GFP was hardly impaired on fusion with BCCPDelta100 and biotinylation of the latter. PMID- 19560435 TI - Precursor ion scanning and sequencing of arginine-ADP-ribosylated peptide by mass spectrometry. AB - Arginine (Arg)-specific ADP-ribosylation is one of the posttranslational modifications of proteins and is thought to play an important role in reversibly regulating functions of the target proteins in eukaryotes. However, the physiological target protein has not been established. We examined the fragmentation pattern of both ADP-ribosyl-Arg (ADP-R-Arg) and Arg-ADP-ribosylated peptides by quadrupole tandem mass spectrometry and found a specific cleavage of ADP-R-Arg into N-(ADP-ribosyl)-carbodiimide (ADP-R-carbodiimide) and ornithine. Based on this specific fragmentation pattern, we successfully identified the modification site and sequence of Arg-ADP-ribosylated peptide using a two-step collision and showed that ADP-R-carbodiimide is an excellent marker ion for precursor ion scanning of Arg-ADP-ribosylated peptide. We propose that a combination of the precursor ion scanning with ADP-R-carbodiimide as a marker ion and two-step collision is useful in searching for physiological target proteins of Arg-ADP-ribosylation. PMID- 19560437 TI - Bidirectional regulation of insulin receptor autophosphorylation and kinase activity by peroxynitrite. AB - Accumulating evidence suggests that enhanced peroxynitrite formation occurs during diabetes. This report describes the effect of peroxynitrite on insulin receptor (IR) function. Addition of peroxynitrite to purified IR resulted in concentration-dependent tyrosine nitration and thiol oxidation. Interestingly, the basal and insulin-stimulated IR autophosphorylation and tyrosine kinase activity were upregulated at low peroxynitrite concentrations, but downregulated at high peroxynitrite concentrations. Concomitantly, peroxynitrite dramatically reduced (125)I-insulin binding capacity and phosphotyrosine phosphatase activity of IR preparations. Moreover, SIN-1 administration decreased blood glucose levels in normal mice via upregulation of IR/IRS-1 tyrosine phosphorylation. In contrast, SIN-1 markedly increased blood glucose levels in diabetic mice concomitant with downregulation of IR/IRS-1 tyrosine phosphorylation. Taken together, these data provide new insights regarding how peroxynitrite influences IR function in vitro and in vivo, suggesting that peroxynitrite plays a dual role in regulation of IR autophosphorylation and tyrosine kinase activity, and SIN-1 has hyperglycemic effect in diabetic mice. PMID- 19560436 TI - Mixing apples and oranges: Analysis of heterotropic cooperativity in cytochrome P450 3A4. AB - Heterotropic cooperative phenomena have been documented in studies with cytochrome P450 3A4, with few attempts to quantify this behavior other than to show the apparent stimulatory effect of certain CYP3A4 substrates on the enzyme's catalytic activity for others. Here CYP3A4 solubilized in Nanodiscs is studied for its ability to interact with two substrates, alpha-naphthoflavone and testosterone, which produce transitions in the heme spin state with apparent spectral affinities (corrected for membrane partitioning) of 7 and 38 microM, respectively. Simultaneous addition of both substrates at fixed molar ratios allows for the separation of specific heterotropic cooperative interactions from the simple additive affinities for the given substrate ratios. The absence of any changes in the normalized spectral dissociation constant due to changes in substrate ratio reveals that the observed stimulatory effect is largely due to differences in the relative substrate affinities and the presence of additional substrate in the system, rather than any specific positive heterotropic interactions between the two substrates. PMID- 19560438 TI - Histidine 440 controls the opening of colicin E1 channels in a lipid-dependent manner. AB - The in vitro activity of many pore-forming toxins, in particular, the rate of increase in the membrane conductance induced by the channel-forming domain (P178) of colicin E1 is maximum at an acidic pH. However, after P178 binding at acidic conditions, a subsequent pH shift from 4 to 6 on both sides of the planar bilayer lipid membrane caused a large increase in the trans-membrane current which was solely due to an increase in the number of open channels. This effect required the presence of anionic lipid. Replacing the His440 residue of P178 by alanine eliminated the pH-shift effect thereby showing that it is associated with deprotonation of this histidine residue. It was concluded that alkalinization induced weakening of the electrostatic interactions between colicin and the membrane surface facilitates conformational changes required for the transition of membrane-bound colicin molecules to an active channel state. PMID- 19560439 TI - Na(+)-ATPase in spontaneous hypertensive rats: possible AT(1) receptor target in the development of hypertension. AB - Clinical and experimental data show an increase in sodium reabsorption on the proximal tubule (PT) in essential hypertension. It is well known that there is a link between essential hypertension and renal angiotensin II (Ang II). The present study was designed to examine ouabain-insensitive Na(+)-ATPase activity and its regulation by Ang II in spontaneously hypertensive rats (SHR). We observed that Na(+)-ATPase activity was enhanced in 14-week-old but not in 6-week old SHR. The addition of Ang II from 10(-12) to 10(-6) mol/L decreased the enzyme activity in SHR to a level similar to that obtained in WKY. The Ang II inhibitory effect was completely reversed by a specific antagonist of AT(2) receptor, PD123319 (10(-8) mol/L) indicating that a system leading to activation of the enzyme in SHR is inhibited by AT(2)-mediated Ang II. Treatment of SHR with losartan for 10 weeks (weeks 4-14) prevents the increase in Na(+)-ATPase activity observed in 14-week-old SHR. These results indicate a correlation between AT(1) receptor activation in SHR and increased ouabain-insensitive Na(+)-ATPase activity. Our results open new possibilities towards our understanding of the pathophysiological mechanisms involved in the increased sodium reabsorption in PT found in essential hypertension. PMID- 19560441 TI - A novel nitric oxide synthase expressed specifically in the olfactory center. AB - Nitric oxide (NO) plays important roles in the olfactory center of various animals. In the terrestrial slug, NO is indispensable for field potential oscillation in the higher olfactory center, the procerebrum (PC), and also for odor learning. Here we identify a novel NO synthase (NOS) gene, limNOS2, in the terrestrial slug. The mRNA (approximately 10kb) of limNOS2 encodes a protein consisting of 1616 amino acids, including a PDZ domain. The protein has 70.0% sequence identity with the previously identified limNOS1 gene. In contrast to limNOS1, however, limNOS2 is expressed specifically in the PC. Moreover, most of the cells in the PC contain limNOS2 mRNA, indicating that the nonbursting neurons, the major constituent of the PC, have this mRNA. The expression pattern of limNOS2 conforms well to the pattern of NOS enzymatic histochemical staining. Our present findings indicate that limNOS2 is responsible for most of the NO generation in the PC. PMID- 19560440 TI - Pore size of swelling-activated channels for organic osmolytes in Jurkat lymphocytes, probed by differential polymer exclusion. AB - The present study explores the impact of the molecular size on the permeation of low-molecular-weight polyethylene glycols (PEG200-1500) through the plasma membrane of Jurkat cells under iso- and hypotonic conditions. To this end, we analyzed the cell volume responses to PEG-substituted solutions of different osmolalities (100-300 mOsm) using video microscopy. In parallel experiments, the osmotically induced changes in the membrane capacitance and cytosolic conductivity were measured by electrorotation (ROT). Upon moderate swelling in slightly hypotonic solutions (200 mOsm), the lymphocyte membrane remained impermeable to PEG300-1500, which allowed the cells to accomplish regulatory volume decrease (RVD). During RVD, lymphocytes released intracellular electrolytes through the swelling-activated pathways, as proved by a decrease of the cytosolic conductivity measured by electrorotation. RVD also occurred in strongly hypotonic solutions (100 mOsm) of PEG600-1500, whereas 100 mOsm solutions of PEG300-400 inhibited RVD in Jurkat cells. These findings suggest that extensive hypotonic swelling rendered the cell membrane highly permeable to PEG300-400, but not to PEG600-1500. The swelling-activated channels conducting PEG300-400 were inserted into the plasma membrane from cytosolic vesicles via swelling-mediated exocytosis, as suggested by an increase of the whole cell capacitance. Using the hydrodynamic radii R(h) of PEGs (determined by viscosimetry), the observed size-selectivity of membrane permeation yielded an estimate of approximately 0.74 nm for the cut-off radius of the swelling activated channel for organic osmolytes. Unlike PEG300-1500, the smallest PEG (PEG200, R(h)=0.5 nm) permeated the lymphocyte membrane under isotonic conditions thus leading to a continuous isotonic swelling. The results are of interest for biotechnology and biomedicine, where PEGs are widely used for cryopreservation of cells and tissues. PMID- 19560442 TI - FATP1 mediates fatty acid-induced activation of AMPK in 3T3-L1 adipocytes. AB - Fatty acid transport proteins are integral membrane acyl-CoA synthetases implicated in adipocyte fatty acid influx and esterification. FATP-dependent production of AMP was evaluated using FATP4 proteoliposomes, and fatty acid dependent activation of AMP-activated protein kinase (AMPK) was assessed in 3T3 L1 adipocytes. Insulin-stimulated fatty acid influx (palmitate or arachidonate) into cultured adipocytes resulted in an increase in the phosphorylation of AMPK and its downstream target acetyl-CoA carboxylase. Consistent with the activation of AMPK, palmitate uptake into 3T3-L1 adipocytes resulted in an increase in intracellular [AMP]/[ATP]. The fatty acid-induced increase in AMPK activation was attenuated in a cell line expressing shRNA targeting FATP1. Taken together, these results demonstrate that, in adipocytes, insulin-stimulated fatty acid influx mediated by FATP1 regulates AMPK and provides a potential regulatory mechanism for balancing de novo production of fatty acids from glucose metabolism with influx of preformed fatty acids via phosphorylation of acetyl-CoA carboxylase. PMID- 19560444 TI - Interaction of porphyrins with human organic anion transporting polypeptide 1B1. AB - The existence of a porphyrin uptake transporter in hepatocytes has been hypothesized in recent years, but to date it has not been identified. While the linear tetrapyrrole bilirubin has been shown to be a substrate for the organic anion transporting polypeptide 1B1 (OATP1B1), similar studies have not been conducted for the cyclic tetrapyrroles (porphyrins). The aim of this study was to determine the structural features of linear and cyclic tetrapyroles necessary for interaction with OATP1B1. The interaction was quantified using HEK cells stably expressing OATP1B1 and measuring the inhibition of OATP1B1-mediated uptake of estradiol 17beta-d-glucuronide in the presence or absence of various linear and cyclic tetrapyrroles. Ditaurine-conjugated bilirubin was the most potent inhibitor of uptake, with an IC50 of 5 nM, while the substitution of the taurine side chains with methyl ester eliminated the inhibition of estradiol 17beta-d glucuronide uptake. Hematoporphyrin, a cyclic tetrapyrrole with carboxyalcohol side chains at positions C-3 and C-8 and carboxyethyl side chains at positions 13 and 17 had an IC50 of 60 nM, while porphyrins lacking charged side chains such as etioporphyrin I and phthalocyanine did not inhibit OATP1B1. Chlorin e6 and hematoporphyrin were shown to be competitive inhibitors of OATP1B1-mediated uptake of bromosulfophthalein with Kis of 5.8+/-0.3 and 1.6+/-0.3 microM, respectively. While these studies do not provide direct evidence, they do support the assumption that tetrapyrroles are transported by OATP1B1. Additionally, these findings offer a possible explanation for the clinical observation that patients suffering from certain porphyrietic diseases have a reduced ability to excrete organic anions. PMID- 19560443 TI - Aerobic fitness is associated with gray matter volume and white matter integrity in multiple sclerosis. AB - Alterations in gray and white matter have been well documented in individuals with multiple sclerosis. Severity and extent of such brain tissue damage have been associated with cognitive impairment, disease duration and neurological disability, making quantitative indices of tissue damage important markers of disease progression. In this study, we investigated the association between cardiorespiratory fitness and measures of gray matter atrophy and white matter integrity. Employing voxel-based approaches to analysis of gray matter and white matter, we specifically examined whether higher levels of fitness in multiple sclerosis participants were associated with preserved gray matter volume and integrity of white matter. We found a positive association between cardiorespiratory fitness and regional gray matter volumes and higher focal fractional anisotropy values. Statistical mapping revealed that higher levels of fitness were associated with greater gray matter volume in the midline cortical structures including the medial frontal gyrus, anterior cingulate cortex and the precuneus. Further, we also found that increasing levels of fitness were associated with higher fractional anisotropy in the left thalamic radiation and right anterior corona radiata. Both preserved gray matter volume and white matter tract integrity were associated with better performance on measures of processing speed. Taken together, these results suggest that fitness exerts a prophylactic influence on the structural decline observed early on, preserving neuronal integrity in multiple sclerosis, thereby reducing long-term disability. PMID- 19560445 TI - Miscibility and phase behavior of DPPG and perfluorocarboxylic acids at the air water interface. AB - The miscibility and phase behavior of two components of phospholipids and perfluorocarboxylic acids [FCn; perfluorododecanoic acid (FC12), perfluorotetradecanoic acid (FC14), perfluorohexadecanoic acid (FC16), and perfluorooctadecanoic acid (FC18)] have been systematically investigated using Langmuir monolayer technique. Dipalmitoylphosphatidylglycerol (DPPG) is utilized as a phospholipid component in biomembranes. Surface pressure (pi)-molecular area (A) and surface potential (DeltaV)-A isotherms have been measured for the DPPG/FCn systems on 0.15 M NaCl (pH 2.0) at 298.2K. From the isotherm results, two-dimensional phase diagrams are constructed and classified into miscible and immiscible patterns. Furthermore, the phase behavior of the DPPG/FCn systems has been morphologically examined using fluorescence microscopy (FM) and atomic force microscopy (AFM). These images indicate different phases among the four systems. In particular, specific phase morphology is observed in the middle molar fraction range for the DPPG/FC14 system; FC14 is selectively excluded from mixed DPPG-FC14 monolayers to be concentrated in the phase boundary as surface pressure increases. Then DPPG is refined as a patched film. Moreover, the data obtained here are compared to those in the previous systems in which different kinds of phospholipids were treated. Through a series of the miscibility investigations, it is proposed that combinations of hydrophobic chain lengths and of polar headgroups contribute to the monolayer miscibility between phospholipids and perfluorocarboxylic acids. PMID- 19560446 TI - Effects of NAT2 polymorphism on SASP pharmacokinetics in Chinese population. AB - BACKGROUND: Sulfasalazine (SASP) pharmacologic actions are widely applied in clinical therapy. The role of N-Acetyltransferase 2 (NAT2) in the pharmacokinetics of SASP and its metabolites has not been clarified. We investigated the effects of genetic polymorphism of NAT2 on pharmacokinetic profiles of SASP and its two metabolites, sulfapyridine (SP) and N acetylsufapyridine (AcSP). METHODS: Eighteen subjects were recruited and divided into 3 groups by NAT2 genotype: wild type (w/w), heterozygous variant (w/m), homozygous variant (m/m). After taking 1000mg SASP tablets, the plasma concentrations of SASP, SP and AcSP were measured with HPLC method and pharmacokinetic parameters were calculated by using the computing program 3P97. RESULTS: The AUC(0)(-)(72) and Cmax of SP in m/m subjects were significantly higher than those in w/m and w/w subjects, with the values of 172.57+/-49.42, 103.38+/-39.85, 71.37+/-17.52mg h/l, and 9.65+/-2.34, 6.10+/-1.79, 4.55+/ 1.38mg/l, respectively. In contrast, the AUC(0)(-)(72) of AcSP was significantly lower in m/m subjects. The Cmax of AcSP in w/w, w/m and m/m subjects was 12.67+/ 3.32, 9.07+/-2.29 and 4.22+/-0.93mg/l, respectively, with significant differences among groups. However, there was no significant difference in any pharmacokinetic parameter of SASP among groups. CONCLUSION: Different NAT2 genotypes, leading to functional heterogeneity of NAT2, may affect pharmacokinetics of SP and AcSP. Therefore, genotyping NAT2 gene before administration would be important in SASP therapy. PMID- 19560447 TI - BMI and hyperinsulinemia in children. AB - OBJECTIVES: Overweight is associated with hyperinsulinemia in adults and children. The aim of our study was to test if body mass index (BMI) alone or in combination with waist circumference (WC) predicts hyperinsulinemia in individual children. METHODS: In 466 healthy German schoolchildren aged 7 to 12 years enrolled in the population based PEP Family Heart Study, anthropometric data, standard laboratory data and fasting serum insulin concentrations were assessed. RESULTS: Among children with hyperinsulinemia (fasting serum insulin concentrations>85th age specific percentile), 56% were not overweight (BMI-for age<85th percentile). Among overweight children, 54% were normoinsulinemic. Increased waist circumference (WC for age and gender>85th percentile) was not a better predictor of hyperinsulinemia, neither alone nor in combination with BMI. CONCLUSION: Overweight and abdominal obesity are poor indicators of hyperinsulinemia in individual German school children aged 7 to 12 years. Insulin measurement seems to be necessary to reliably detect hyperinsulinemia. PMID- 19560448 TI - Antioxidant status and lipid peroxidation in small intestinal mucosa of children with celiac disease. AB - OBJECTIVE: To explain the role of oxidative stress in the pathology of celiac disease. DESIGN AND METHODS: The activities of antioxidant enzymes and the levels of glutathione and lipid hydroperoxides were measured in the samples of small intestinal biopsies from 39 children with different forms of the disease and in 19 control subjects. RESULTS: The activities of analyzed enzymes varied significantly between the examined groups. An increase in the activities of superoxide dismutase was observed in patients with active and silent celiac disease, while the activities of glutathione peroxidase and glutathione reductase and the glutathione content were significantly reduced. The level of lipid hydroperoxides was significantly elevated in these groups. CONCLUSIONS: Oxidative stress is an important factor in the pathogenesis of celiac disease. The antioxidant capacity of celiac patients is significantly reduced, mostly by a depletion of glutathione. Natural antioxidants and appropriate dietary supplements could be important complements to the classic therapy of celiac disease. PMID- 19560449 TI - The effect of anticoagulants on the quality and biological efficacy of platelet rich plasma. AB - OBJECTIVES: This study investigated the effect of anticoagulants on platelet-rich plasma (PRP) quality to determine the appropriate anticoagulants for PRP production. DESIGN AND METHODS: This study was carried out at the Plastic Surgery Hospital of Peking Union Medical College. The microstructure of platelets collected with heparin, citrate, acid citrate dextrose (ACD) and citrate theophylline-adenosine-dipyridamole (CTAD) was observed. The extent of spontaneous activation of platelets was detected by measuring sP-selectin in plasma. The amount of TGF-beta1 released from PRP and the effect of PRP on cell proliferation were also studied. RESULTS: ACD and CTAD were superior to heparin and citrate in maintaining the integrity of platelet structures and preventing the platelet spontaneous activation. ACD-PRP and CTAD-PRP released more TGF-beta1 and significantly enhanced the proliferation of human marrow stromal cells compared to heparin-PRP and citrate-PRP. CONCLUSIONS: The PRP quality was closely related to the type of anticoagulants. ACD and CTAD are appropriate anticoagulants for PRP production. PMID- 19560450 TI - Comparison of immunoturbidimetric and immunonephelometric assays for specific proteins. AB - Immunoturbidimetric assays for specific proteins are available on "open system" clinical chemistry analyzers. The analytical performance of nine immunoturbidimetric specific protein assays (C3, C4, CRP, Haptoglobin, IgA, IgG, IgM, RF, and Transferrin) was compared to immunonephelometry. Testing was performed on the Abbott ARCHITECT ci8200 and the Dade Behring BNII nephelometer and evaluated for precision, linearity, limit of detection, prozone phenomenon, method comparison, workflow, and proficiency testing survey comparison. Immunoturbidimetric assays performance was satisfactory for total precision, linearity, limit of detection and the prozone effect was not observed. Method comparison was acceptable for the immunoglobulins, CRP and transferrin but less favorable for the other assays, likely due to methodology and antibody specificity differences. Immunourbidimetric specific protein assays allow for efficient test consolidation on a general purpose clinical chemistry analyzer. PMID- 19560451 TI - A novel ultra-sensitive enzyme immunoassay for soluble human insulin receptor ectodomain and its measurement in urine from healthy subjects and patients with diabetes mellitus. AB - OBJECTIVE: For the early identification of patients at risk of developing diabetes mellitus, and to prevent the onset of diabetes by performing dietary counseling and exercise guidance, we have developed an ultra-sensitive immune complex transfer enzyme immunoassay (ICT-EIA) to measure soluble human insulin receptor ectodomain (sIRalpha) in urine which is collected non-invasively. DESIGN AND METHODS: We developed ICT-EIA for sIRalpha and measured urinary sIRalpha from 106 healthy volunteers, 35 obese volunteers and 42 patients with diabetes. RESULTS: The detection limit of ICT-EIA (0.04 pg/mL), using a urine sample of as little as 100 microL, was a few hundred-fold higher than that of conventional ELISA. Using ICT-EIA, the urinary sIRalpha level in patients with diabetes (9.7+/ 20.1 pg/mg creatinine) was significantly higher than those in healthy volunteers (1.4+/-0.9; P<0.001). CONCLUSION: ICT-EIA for sIRalpha may be useful as a good marker for evaluating diabetes risk. PMID- 19560453 TI - The kallikrein family of proteins as urinary biomarkers for the detection of prostate cancer. AB - BACKGROUND: Several urinary biomarkers have been assessed as showing a discriminatory ability to differentially diagnose prostate cancer, albeit with manipulation of the prostate. Here we examine the clinical utility of multiple members of the kallikrein family of proteins in non-manipulative urinary biomarker testing. METHODS: Forty urine samples were collected from patients admitted for urological examination. Twenty, with a confirmed benign diagnosis and 20 with prostate cancer. The levels of 14 kallikrein proteins were measured in patient's urine and normalized for creatinine. RESULTS: Ten of the 14 kallikreins tested had detectable levels in urine. However, none showed statistical significance in discriminating patients. Serum PSA was superior to urine PSA and other urinary kallikreins in separating patients with and without prostate cancer. CONCLUSIONS: We were unable to distinguish men with and without prostate cancer using multiple kallikreins as urinary biomarkers. These results highlight the difficulties in diagnosing prostate cancer via urine testing for soluble biomarkers. PMID- 19560452 TI - Prognostic significance of adipocytokines and extracellular matrix activity in heart failure patients with high B-type natriuretic peptide. AB - OBJECTIVES: To perform risk stratification by serum adipocytokines and serum markers of extracellular matrix in heart failure patients with high b-type natriuretic peptide (BNP). METHODS: Patients with heart failure were enrolled in this study. Serum adipocytokines and serum markers of extracellular matrix were analyzed. RESULTS: In total, 131 patients were enrolled and followed-up for 240+/ 174 days. Mortality was significantly associated with adiponectin, resistin, type III amioterminal propeptide of procollagen (PIIINP), matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase 1 (TIMP-1), and serum creatinine. Heart failure-related admission was significantly associated with apelin, and PIIINP. Cox regression analysis identified that mortality and heart failure related admissions were significantly associated with MMP-2 (P=0.008) and PIIINP (P=0.011), respectively. CONCLUSIONS: Serum markers of extracellular matrix rather than adipocytokines may warrant further risk stratification for impacting the prognosis of patients with heart failure with high BNP. PMID- 19560454 TI - Lipopolysaccharide-binding protein (LBP) gene polymorphisms: rapid genotyping by real-time PCR and association with infective endocarditis. AB - OBJECTIVES: Lipopolysaccharide-binding protein (LBP) was recently demonstrated to be a supportive biomarker for the diagnostic and therapeutic monitoring of infective endocarditis (IE) with a considerable variability in the individual LBP response of IE patients. DESIGN AND METHODS: LBP was measured by chemiluminescence assay in sera from 78 IE patients. Moreover, three new LightCycler PCR assays have been developed for sequence variation analysis and the distribution of the five LBP polymorphisms c.-1978C>T, c.-836T>C, c.291C>T, c.613A>G and c.1306C>T was determined in IE patients and healthy blood donors. RESULTS: A weak association with IE was determined for the two single nucleotide polymorphisms c.291C>T and c.613A>G; the frequencies of the other polymorphisms did not differ significantly between IE patients and controls. Elevated serum LBP concentrations of infective patients did not correlate with genotypes. CONCLUSIONS: The association of the polymorphisms c.291C>T and c.613A>G suggest a role of LBP in the disease manifestation of IE. PMID- 19560456 TI - Clarithromycin suppresses airway hyperresponsiveness and inflammation in mouse models of asthma. AB - Macrolide antibiotics, a class of potent antimicrobials, also possess immunomodulatory/anti-inflammatory properties. These properties are considered fundamental for the efficacy of macrolide antibiotics in the treatment of diffuse panbronchiolitis and cystic fibrosis. In patients with asthma, macrolide antibiotics have been reported to reduce airway hyperresponsiveness and improve pulmonary function. However, their beneficial actions in asthmatics possibly could be attributed to antimicrobial activity against atypical pathogens (e.g. Chlamydia pneumoniae), corticosteroid-sparing effect (inhibition of exogenous corticosteroid metabolism), and/or their anti-inflammatory/immunomodulatory effects. In order to investigate whether efficacy of macrolide antibiotics in asthma results from their immunomodulatory/anti-inflammatory activity, the influence of clarithromycin pretreatment (2 h before challenge) was examined on ovalbumin-induced airway hyperresponsiveness and airway inflammation in the mouse. Clarithromycin treatment (200 mg/kg intraperitoneally) decreased IL-4, IL 5, IL-13, CXCL2 and CCL2 concentrations in bronchoalveolar lavage fluid and markedly reduced inflammatory cell accumulation in bronchoalveolar lavage fluid and into the lungs, as revealed by histopathological examination. Furthermore, clarithromycin-induced reduction in inflammation was accompanied by normalization of airway hyperresponsiveness. In summary, in ovalbumin-induced mouse models, clarithromycin efficiently inhibited two important pathological characteristics of asthma, airway hyperresponsiveness and inflammation. These data suggest that the efficacy of clarithromycin, as well as of other macrolide antibiotics, in asthmatic patients could be attributed to their anti inflammatory/immunomodulatory properties, and not only to their antimicrobial activity or exogenous corticosteroid-sparing effects. PMID- 19560455 TI - Phosphorylation and consequent stimulation of the tyrosine kinase c-Abl by PKA in mouse spermatozoa; its implications during capacitation. AB - Upon ejaculation, spermatozoa undergo a series of post-translational modifications in a process known as capacitation in order to prepare for fertilization. In the absence of capacitation, fertilization cannot occur. Spermatozoa are unusual in that one of the hallmarks of capacitation is a global up-regulation in phosphotyrosine expression, which is known to be mediated upstream by PKA. Little is known about the signaling events downstream of PKA apart from the involvement of SRC, as a key mediator of PKA-induced tyrosine phosphorylation in the sperm tail. Here we describe the presence of c-Abl in mouse spermatozoa. In vitro analysis confirmed that PKA can up-regulate c-Abl kinase activity. In vivo, this tyrosine kinase was found to associate, and become threonine phosphorylated by PKA in the sperm flagellum. By treating spermatozoa with hemolysin we could demonstrate that a significant proportion of the tyrosine phosphorylation associated with capacitation could be suppressed by the c-Abl inhibitor, Gleevac. This is the first report of c-Abl being up-regulated by PKA for any cell type. We present a model, whereby these kinases may operate together with SRC to ensure optimal levels of tyrosine phosphorylation in the sperm flagellum during the attainment of a capacitated state. PMID- 19560457 TI - Interference RNA (RNAi)-based silencing of endogenous thrombopoietin receptor (Mpl) in Dami cells resulted in decreased hNUDC-mediated megakaryocyte proliferation and differentiation. AB - Recently our laboratory reported evidence showing that hNUDC acts as an additional cytokine for thrombopoietin receptor (Mpl). Previously known as the human homolog of a fungal nuclear migration protein, hNUDC plays a critical role in megakaryocyte differentiation and maturation. Here we sought to further clarify the hNUDC-Mpl ligand-receptor relationship by utilizing interference RNA (RNAi) to knockdown Mpl expression in a megakaryocyte cell line. We created U6 promoter driven constructs to express short hairpin RNAs (shRNA) with affinity for different sites on Mpl mRNA. By including Mpl-EGFP fusion protein in these constructs, we were able to effectively screen the shRNA that was most efficient in inhibiting Mpl mRNA expression. This shRNA was subsequently transferred into a lentivirus vector and transduced into Dami cells, a cell line which constitutively expresses endogenous Mpl. This lentiviral vector was also designed to simultaneously express EGFP to monitor transfection efficiency. Our results show that lentivirus can be used to effectively deliver shRNAs into Dami cells and cause specific inhibition of Mpl protein expression after transduction. Furthermore, we show the functional effects of shRNA-mediated Mpl silencing by demonstrating reduced hNUDC stimulated megakaryocyte proliferation and differentiation. Thus, the use of a RNAi knockdown strategy has allowed us to pinpoint the connection of hNUDC with Mpl in the regulation of megakaryocyte maturation. PMID- 19560458 TI - A novel Drosophila Girdin-like protein is involved in Akt pathway control of cell size. AB - The Akt signaling pathway is well known to regulate cell proliferation and growth. Girdin, a novel substrate of Akt, plays a crucial role in organization of the actin cytoskeleton and cell motility under the control of Akt. We here identified a novel Girdin-like protein in Drosophila (dGirdin), which has two isoforms, dGirdin PA and dGirdin PB. dGirdin shows high homology with human Girdin in the N-terminal and coiled-coil domains, while diverging at the C terminal domain. On establishment of transgenic fly lines, featuring knockdown or overexpression of dGirdin in vivo, overexpression in the wing disc cells induced ectopic apoptosis, implying a role in directing apoptosis. Knockdown of dGirdin in the Drosophila wing imaginal disc cells resulted in reduction of cell size. Furthermore, this was enhanced by half reduction of the Akt gene dose, suggesting that Akt positively regulates dGirdin. In the wing disc, cells in which dGirdin was knocked down exhibited disruption of actin filaments. From these in vivo analyses, we conclude that dGirdin is required for actin organization and regulation of appropriate cell size under control of the Akt signaling pathway. PMID- 19560459 TI - GM-CSF regulates the ERK1/2 pathways and protects injured retinal ganglion cells from induced death. AB - Granulocyte-macrophage-colony-stimulating-factor (GM-CSF) is a potent hematopoietic cytokine. In the present study, we examined whether GM-CSF is neuroprotective in retinal ganglion cells (RGCs). First, we studied the expression of GM-CSF and the GM-CSF-alpha-receptor in rat and human retina and in RGC-5 cells. Then, RGC-5 cells were incubated with apoptosis-inducing agents (e.g., staurosporine, glutamate and NOR3). The cell death was assessed by Live Death-Assays and apoptosis-related-proteins were examined by immunoblotting. In addition, the expression of phosphorylated ERK1/2-pathway-proteins after incubation with GM-CSF and after inhibiting MEK1/2 with U0126 was analyzed. To assess the in vivo-effect, first staurosporine or GM-CSF plus staurosporine was injected into the vitreous body of Sprague-Dawley rats. In a second axotomy model the optic nerve was cut and GM-CSF was injected into the vitreous body. In both models, the RGCs were labeled retrogradely with either Fluoro-Gold or 4-Di-10-Asp and counted. As a first result, we identified GM-CSF and the GM-CSF-alpha receptor in rat and human retina as well as in RGC-5 cells. Then, in the RGC-5 cells GM-CSF counteracts induced cell death in a dose-and time-dependent manner. With respect to apoptosis, Western blot analysis revealed a decreased Bad expression and an increased Bcl-2-expression after co-incubation with GM-CSF. Concerning signaling pathways, incubation with GM-CSF activates the ERK1/2 pathway, whereas inhibition of MEK1/2 with U0126 strongly decreased the phosphorylation downstream in the ERK1/2 pathway, and the antiapoptotic activity of GM-CSF in vitro. Like in vitro, GM-CSF counteracts the staurosporine-induced cell death in vivo and protects RGCs from axotomy-induced degeneration. Our data suggest that GM-CSF might be a novel therapeutic agent in neuropathic disease of the eye. PMID- 19560460 TI - Large-scale purification of ribosome-nascent chain complexes for biochemical and structural studies. AB - Here we present a method to purify large amounts of highly pure and stably arrested ribosome-nascent chain complexes (RNCs) from Escherichia coli cells. It relies on the combined use of translation-arrest sequences to generate nascent polypeptides of specified length and subsequent tag purification of the RNCs. Moreover, we adapted this method for the in vivo production of RNCs with specific isotope labeling of the nascent chains for nuclear magnetic resonance (NMR) studies. This method opens therefore possibilities for a wide range of biochemical and structural studies exploring conformations of nascent chains during the early steps of protein folding and targeting. PMID- 19560461 TI - Syndecan-4 promotes the retention of phosphatidylinositol 4,5-bisphosphate in the plasma membrane. AB - Although phosphatidylinositol 4,5-bisphosphate (PIP(2)) regulates syndecan-4 function, the potential influence of syndecan-4 on PIP(2) remains unknown. GFP containing PIP(2)-binding-PH domain of phospholipase Cdelta (GFP-PHdelta) was used to monitor PIP(2). Syndecan-4 overexpression in COS-7 cells enhanced membrane translocation of GFP-PHdelta, while the opposite was observed when syndecan-4 was knocked-down. PIP(2) levels were higher in total phospholipids extracted from rat embryo fibroblasts expressing syndecan-4. Syndecan-4-induced membrane targeting of GFP-PHdelta was further enhanced by phosphoinositide-3 kinase inhibitor, but not by phospholipase C (PLC) inhibitor. Besides, both ionomycin and epidermal growth factor caused dissociation of GFP-PHdelta from plasma membrane, an effect that was significantly delayed by syndecan-4 over expression. Collectively, these data suggest that syndecan-4 promotes plasma membrane retention of PIP(2) by negatively regulating PLC-dependent PIP(2) degradation. PMID- 19560462 TI - Methylene blue and dimebon inhibit aggregation of TDP-43 in cellular models. AB - Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitinated inclusions (FTLD-U) are major neurodegenerative diseases with TDP 43 pathology. Here we investigated the effects of methylene blue (MB) and dimebon, two compounds that have been reported to be beneficial in phase II clinical trials of Alzheimer's disease (AD), on the formation of TDP-43 aggregates in SH-SY5Y cells. Following treatment with 0.05 microM MB or 5 microM dimebon, the number of TDP-43 aggregates was reduced by 50% and 45%, respectively. The combined use of MB and dimebon resulted in a 80% reduction in the number. These findings were confirmed by immunoblot analysis. The results indicate that MB and dimebon may be useful for the treatment of ALS, FTLD-U and other TDP-43 proteinopathies. PMID- 19560463 TI - A contact-waiting-time metric and RNA folding rates. AB - Metrics for indirectly predicting the folding rates of RNA sequences are of interest. In this letter, we introduce a simple metric of RNA structural complexity, which accounts for differences in the energetic contributions of RNA base contacts toward RNA structure formation. We apply the metric to RNA sequences whose folding rates were previously determined experimentally. We find that the metric has good correlation (correlation coefficient: -0.95, p<<0.01) with the logarithmically transformed folding rates of those RNA sequences. This suggests that the metric can be useful for predicting RNA folding rates. We use the metric to predict the folding rates of bacterial and eukaryotic group II introns. Future applications of the metric (e.g., to predict structural RNAs) could prove fruitful. PMID- 19560464 TI - Stability of Helicobacter pylori CagA oncoprotein in human gastric epithelial cells. AB - Upon delivery into gastric epithelial cells, Helicobacter pylori cytotoxin associated gene A (CagA) binds and deregulates cellular proteins such as Src homology 2 domain-containing protein tyrosine phosphatase 2 and partitioning defective 1 (PAR1), thereby acting as an epigenetic oncoprotein that promotes early phases of gastric cancer development. To elucidate the spatial and temporal contribution of CagA to carcinogenesis, it is crucial to know the stability of CagA in host cells. Here we show that the biological half-life of CagA is about 200 min in gastric epithelial cells. Furthermore, deletion of the PAR1-binding sequence accelerates CagA degradation. Thus, CagA is a relatively short half-life protein whose stability may be modulated through complex formation with PAR1. PMID- 19560465 TI - FoxO4 inhibits NF-kappaB and protects mice against colonic injury and inflammation. AB - BACKGROUND & AIMS: FoxO4 is a member of the forkhead box transcription factor O (FoxO) subfamily. FoxO proteins are involved in diverse biological processes. In this study, we examine the role of FoxO4 in intestinal mucosal immunity and inflammatory bowel disease (IBD). METHODS: Foxo4-null mice were subjected to trinitrobenzene sulfonic acid (TNBS) treatment. Microarray analysis and quantitative reverse transcription polymerase chain reaction were used to identify the cytokine transcripts that were altered by Foxo4 deletion. The effects of Foxo4 deficiency on the intestinal epithelial permeability and levels of tight junction proteins were examined by permeable fluorescent dye and Western blot. The molecular and cellular mechanisms by which FoxO4 regulates the mucosal immunity were explored through immunologic and biochemical analyses. The expression level of FoxO4 in intestinal epithelial cells of patients with IBD was examined with immunohistochemistry. RESULTS: Foxo4-null mice were more susceptible to TNBS injury-induced colitis. The chemokine CCL5 is significantly up-regulated in the colonic epithelial cells of Foxo4-null mice, with increased recruitment of CD4(+) intraepithelial T cells and up-regulation of cytokines interferon-gamma and tumor necrosis factor-alpha in the colon. Foxo4 deficiency also resulted in an increase in intestinal epithelial permeability and down regulation of the tight junction proteins ZO-1 and claudin-1. Mechanistically, FoxO4 inhibited the transcriptional activity of nuclear factor-kappaB (NF kappaB), and Foxo4 deficiency is associated with increased NF-kappaB activity in vivo. FoxO4 transcription is transiently repressed in response to TNBS treatment and in patients with IBD. CONCLUSION: These results indicate that FoxO4 is an endogenous inhibitor of NF-kappaB and identify a novel function of FoxO4 in the regulation of NF-kappaB-mediated mucosal immunity. PMID- 19560467 TI - Multiplex cytokine profiling with highly pathogenic material: use of formalin solution in luminex analysis. AB - Work with highly pathogenic material mandates the use of biological containment facilities, involving microbiological safety cabinets and specialist laboratory engineering structures typified by containment level 3 (CL3) and CL4 laboratories. Consequences of working in high containment are the practical difficulties associated with containing specialist assays and equipment often essential for experimental analyses. In an era of increased interest in biodefence pathogens and emerging diseases, immunological analysis has developed rapidly alongside traditional techniques in virology and molecular biology. For example, in order to maximise the use of small sample volumes, multiplexing has become a more popular and widespread approach to quantify multiple analytes simultaneously, such as cytokines and chemokines. The luminex microsphere system allows for the detection of many cytokines and chemokines in a single sample, but the detection method of using aligned lasers and fluidics means that samples often have to be analysed in low containment facilities. In order to perform cytokine analysis in materials from high containment (CL3 and CL4 laboratories), we have developed an appropriate inactivation methodology after staining steps, which although results in a reduction of median fluorescent intensity, produces statistically comparable outcomes when judged against non-inactivated samples. This methodology thus extends the use of luminex technology for material that contains highly pathogenic biological agents. PMID- 19560466 TI - Chronic treatment with estrogen receptor agonists restores acquisition of a spatial learning task in young ovariectomized rats. AB - Previous work has shown that continuous estradiol replacement in young ovariectomized rats enhances acquisition of a delayed matching-to-position (DMP) T-maze task over that of ovariectomized controls. The mechanism by which estradiol confers this benefit has not been fully elucidated. This study examined the role of selective estrogen receptor agonists of ERalpha, ERbeta, and GPR30 in the enhancement of spatial learning on a DMP task by comparing continuous estradiol replacement with continuous administration of PPT (an agonist of ERalpha), DPN (an agonist of ERbeta), or G-1 (an agonist of GPR30) relative to gonadally intact and ovariectomized vehicle-treated controls. It was found that ovariectomy impaired acquisition on this task, whereas all ER selective agonists restored the rate of acquisition to that of gonadally intact controls. These data suggest that estradiol can work through any of several estrogen receptors to enhance the rate of acquisition on this task. PMID- 19560468 TI - Self-amplification as a source of interindividual variability: shelter selection in cockroaches. AB - Although group effect and collective decisions have been described in many insect species, the behavioral mechanisms involved in the process remain poorly documented at the individual level. We examined how individual behavior depends on the environmental context and we precisely characterized the behavioral rules leading to settlement of individual cockroaches in resting site. We focused on the spatial and temporal distribution of individuals in absence of conspecifics. Using isolated adult males of the cockroach Periplaneta americana, we showed that the quality of resting sites and the duration of the settlement exerted an influence on the individual decision-making: the probability of leaving a resting site decreased with the time spent under a shelter. A numerical model derived from experimental data suggested that this simple rule of self-amplification can also account for the interindividual variability. PMID- 19560470 TI - Effects of chloride ion binding on the photochemical properties of salinibacter sensory rhodopsin I. AB - Microbial organisms utilize light not only as energy sources but also as signals by which rhodopsins (containing retinal as a chromophore) work as photoreceptors. Sensory rhodopsin I (SRI) is a dual photoreceptor that regulates both negative and positive phototaxis in microbial organisms, such as the archaeon Halobacterium salinarum and the eubacterium Salinibacter ruber. These organisms live in highly halophilic environments, suggesting the possibility of the effects of salts on the function of SRI. However, such effects remain unclear because SRI proteins from H. salinarum (HsSRI) are unstable in dilute salt solutions. Recently, we characterized a new SRI protein (SrSRI) that is stable even in the absence of salts, thus allowing us to investigate the effects of salts on the photochemical properties of SRI. In this study, we report that the absorption maximum of SrSRI is shifted from 542 to 556 nm in a Cl(-)-dependent manner with a K(m) of 307+/-56 mM, showing that Cl(-)-binding sites exist in SRI. The bathochromic shift was caused not only by NaCl but also by other salts (NaI, NaBr, and NaNO(3)), implying that I(-), Br(-), and NO(3)(-) can also bind to SrSRI. In addition, the photochemical properties during the photocycle are also affected by chloride ion binding. Mutagenesis studies strongly suggested that a conserved residue, His131, is involved in the Cl(-)-binding site. In light of these results, we discuss the effects of the Cl(-) binding to SRI and the roles of Cl(-) binding in its function. PMID- 19560469 TI - Translational lessons from scarless healing of cutaneous wounds and regenerative repair of the myocardium. AB - Regenerative healing is the process by which injured tissues are restored to their original structure and function. Many species are capable of healing in this manner. However, in mammals the healing response in most tissues is marked by fibroblast proliferation and scar tissue deposition. While scarring contributes to efficient resolution of mammalian wounds and restoration of at least partial structural and functional support, the final result of scar formation can be more deleterious than the initial insult. This is especially true in the heart, which is sensitive to electrical heterogeneities and altered mechanical properties produced by scarring. Several therapeutic modalities promoting regeneration in skin wounds have been developed that modulate various aspects of the healing process. Targets include cytokine stimulation, control of fibroblast activation, modulation of gap junctions, and stem cell differentiation. Here, we review and compare mechanisms of injury, repair, and scarring in the skin and heart and discuss the promise and caveats of future therapies that may translate to improving repair of myocardial tissues. PMID- 19560471 TI - A model for hormonal control of the menstrual cycle: structural consistency but sensitivity with regard to data. AB - This study presents a 13-dimensional system of delayed differential equations which predicts serum concentrations of five hormones important for regulation of the menstrual cycle. Parameters for the system are fit to two different data sets for normally cycling women. For these best fit parameter sets, model simulations agree well with the two different data sets but one model also has an abnormal stable periodic solution, which may represent polycystic ovarian syndrome. This abnormal cycle occurs for the model in which the normal cycle has estradiol levels at the high end of the normal range. Differences in model behavior are explained by studying hysteresis curves in bifurcation diagrams with respect to sensitive model parameters. For instance, one sensitive parameter is indicative of the estradiol concentration that promotes pituitary synthesis of a large amount of luteinizing hormone, which is required for ovulation. Also, it is observed that models with greater early follicular growth rates may have a greater risk of cycling abnormally. PMID- 19560472 TI - Clinical factors such as B-type natriuretic peptide link to factor VII, endothelial NO synthase and estrogen receptor alpha polymorphism in elderly women. AB - AIMS: This study evaluated the presence of genetic mutations in relation to thrombosis or atherosclerosis in elderly women. MAIN METHODS: This is an observational study of 93 Japanese women with a mean age of 80.9 years recruited from outpatient clinics of Nagoya University and its related hospitals. Ten single nucleotide polymorphisms (SNPs) were studied. Each gene studied acts in or is related to either blood coagulation (factor V Leiden, prothrombin G20210A, factor XIII Val34Leu, factor VII Arg353Gln, MTHFR C677T, beta-fibrinogen G-455A, PAI-1 4G/5G), metabolic syndrome-related pathways (PPARalpha Leu162Val), or endothelium/estrogen system (eNOS Glu298Asp, ERalpha IVS1-401). SNPs were analyzed for their relation to clinical values including lipids, B-type natriuretic peptide (BNP), fasting plasma glucose, tumor necrosis factor-alpha, interleukin-6, cyclic GMP, and nitric oxide metabolites. KEY FINDINGS: Comparisons between the distributions of different genotypes and clinical values showed three relationships. First, factor VII Arg353Gln and HDL-cholesterol (HDL C) were linked to Arg/Arg carriers at higher levels (P=.049). The HDL-C to LDL cholesterol ratio supported this link (P=.027). Second, eNOS Glu298Asp and triglycerides were linked to Glu/Glu carriers at higher levels (P=.031). Third, ERalpha IVS1-401 and BNP were related to CC genotype at lower levels (P=.031). Additionally, the last two relations showed that genotype does not influence the demarcation line of biomarkers, but the plasma/serum levels of biomarkers instead. SIGNIFICANCE: Correlations of factor VII Arg353Gln with HDL-C and eNOS Glu298Asp with triglycerides are new findings. Polymorphisms in the endothelium/estrogen system and the heart failure marker BNP are also correlated, with ERalpha IVS1-401 being the first identified marker. SNPs may be helpful for understanding the pathophysiology of atherosclerotic diseases in elderly women. PMID- 19560473 TI - Oxytocin decreases methamphetamine self-administration, methamphetamine hyperactivity, and relapse to methamphetamine-seeking behaviour in rats. AB - There is emerging evidence that the neuropeptide oxytocin may be utilised as a treatment for various psychopathologies, including drug addictions. Here we used an animal model to assess whether oxytocin might be effective in the treatment of methamphetamine addiction. Sprague-Dawley rats were trained to lever press to intravenously self-administer methamphetamine under a progressive ratio schedule of reinforcement. Once responding had stabilised, one group of rats received escalating doses of oxytocin (0.001, 0.01, 0.1, 0.3, 1 mg/kg) administered intraperitoneally (IP) prior to daily self-administration tests, while other rats received vehicle. After these tests, lever-pressing was extinguished and the ability of methamphetamine primes (IP, 1 mg/kg) to reinstate responding was studied with and without co-administration of oxytocin (IP, 0.3 and 1 mg/kg). Results showed that oxytocin dose-dependently reduced responding for intravenous methamphetamine with an almost complete absence of responding at the highest oxytocin dose (1 mg/kg). Hyperactivity during methamphetamine self-administration was also dose-dependently reduced by oxytocin. Oxytocin (1 but not 0.3 mg/kg) also reduced the ability of methamphetamine to reinstate methamphetamine-seeking behaviour. In separate tests, oxytocin (IP, 0.3 and 1 mg/kg) robustly decreased the hyperactivity and rearing induced by methamphetamine challenge (IP, 1 mg/kg), producing activity levels similar to control animals. This study suggests that oxytocin may have a powerful inhibitory effect on the motivation to consume methamphetamine and on hyperactivity associated with acute methamphetamine intoxication. These results point to the potential utility of human trials of oxytocin as a therapeutic treatment for methamphetamine addiction. PMID- 19560474 TI - Swim stress excitation of nucleus incertus and rapid induction of relaxin-3 expression via CRF1 activation. AB - Relaxin-3 (RLX3), a newly identified member of the relaxin peptide family, is distinguished by its enriched expression in GABA projection neurons of the pontine nucleus incertus (NI), which are postulated to participate in forebrain neural circuits involved in behavioural activation and stress responses. In this regard, corticotrophin-releasing factor-1 receptor (CRF(1)) is abundantly expressed by NI neurons; central CRF administration activates c-fos expression in NI; and various stressors have been reported to increase NI neuron activity. In studies to determine whether a specific neurogenic stressor would activate RLX3 expression, we assessed the effect of a repeated forced swim (RFS) on levels of RLX3 mRNA and heteronuclear (hn) RNA in rat NI by in situ hybridization histochemistry of exon- and intron-directed oligonucleotide probes, respectively. Exposure of rats to an RFS (10 min at 23 degrees C, 24 h apart), markedly increased RLX3 mRNA levels in NI at 30-60 min after the second swim, before a gradual return to basal levels over 2-4 h, while RLX3 hnRNA levels were significantly up-regulated at 60-120 min post-RFS, following a transient decrease at 30 min. Systemic treatment of rats with a CRF(1) antagonist, antalarmin (20 mg/kg, i.p.) 30 min prior to the second swim, blunted the stress-induced effects on RLX3 transcripts. Relative levels of RLX3-immunostaining in NI neurons appeared elevated at 3 h post-swim, but not at earlier time points (30-60 min). These results suggest that acute stress-induced CRF secretion can rapidly alter RLX3 gene transcription by activation of CRF(1) present on NI neurons. More generally, these studies support a role for RLX3 neural networks in the normal neural and physiological response to neurogenic stressors in the rat. PMID- 19560475 TI - Maternal separation interferes with developmental changes in brain vasopressin and oxytocin receptor binding in male rats. AB - Brain vasopressin V(1A) receptors (V(1A)-R) and oxytocin receptors (OT-R) are important modulators of social behaviors. We recently showed that exposure to maternal separation (MS; 3 h daily, postnatal days 1-14) induces changes in social behaviors in juvenile and adult male rats. Here, we hypothesize that MS induces brain region-specific changes in V(1A)-R and OT-R across development, which in turn, may underlie MS-induced changes in social behaviors. We examined the effects of MS on V(1A)-R and OT-R binding in forebrain regions of juvenile (5 weeks), adolescent (8 weeks), and adult (16 weeks) male rats. Robust age-related changes were found for V(1A)-R and OT-R binding in several brain regions. For example, in the lateral septum V(1A)-R binding increased while OT-R binding decreased with age. Most notably, OT-R binding in the caudate putamen showed a 2 fold decrease while OT-R binding in the ventromedial hypothalamus showed a 4-fold increase with age. Importantly, exposure to MS interfered with these developmental changes in several brain regions. Specifically, MS significantly increased V(1A)-R binding in the piriform cortex (at adolescent and adult ages), the lateral septum (at juvenile age), the hypothalamic attack area (at adolescent age), and the dentate gyrus of the hippocampus (at adolescent age), and decreased V(1A)-R binding in the arcuate nucleus (at juvenile age). Moreover, OT-R binding was significantly lower in the agranular cortex (at juvenile and adolescent age), the lateral septum (at adult age) and the caudate putamen (at adult age), but higher in the medial preoptic area (at adolescent age) and ventromedial hypothalamus (at adult age) after exposure to MS. In conclusion, age-dependent changes in V(1A)-R and OT-R binding are likely associated with the maturation of behaviors, such as sexual and aggressive behaviors, while disruption of these changes by MS might contribute to previously observed changes in social behaviors after MS. PMID- 19560476 TI - Impaired control of the oculomotor reflexes in Parkinson's disease. AB - To investigate the role of the basal ganglia in integrating voluntary and reflexive behaviour, the current study examined the ability of patients with Parkinson's disease to voluntarily control oculomotor reflexes. We measured the size of the fixation offset effect (the reduction in saccadic reaction time when a fixation point is removed) during a block of pro- and a block of anti-saccades. Healthy controls showed the expected reduction of the FOE during the anti saccades, which results from efforts to suppress reflexive eye movements (a preparatory set characterized by increased internal control and reduced external control). However, there was no reduction of the FOE in the anti-saccade task in Parkinson's patients, indicating that they are impaired in exerting control over oculomotor reflexes. PMID- 19560478 TI - Ontogeny of fetal movements in the guinea pig. AB - Assessment of fetal motility is an approach to evaluate the development and function of the nervous system before birth. Reference values for the time of first occurrence and the incidence of normal fetal movements are indispensable for studies in which prenatal motor activity is applied as a model to study the central and peripheral nervous systems. Studies on fetal motility have been performed in a few species, particularly in the human. The aim of the present study is to describe the ontogeny of fetal motility in the guinea pig, a precocious polytocous species. After a pilot study to establish procedures for repeated ultasonographic scanning of guinea pigs, 10 domesticated animals were scanned (5.0 or 7.5 MHz convex transducer) at 2-4 day intervals between day 24 and 63 of gestation (term age 68 days). Per animal two selected fetuses were each scanned for 15 min. Images were stored on videotape and analyzed off-line for the first onset, presence and quality of fetal movement patterns, and quantity of sideway bendings, general movements, breathing movements and periods of fetal rest. Twenty-five different movement patterns could be characterized, 6 emerging at the onset of motor activity were performed only temporarily. The very first fetal movement was observed on day 24 gestational age, and subsequently most other movements developed during a period of only 5 days. Interfetal difference in onset of the frequently occurring sideway bendings, general movements, and front and hind limb movements was only 2 days. Sideway bendings and general movements co-existed during days 29 to 43. There were developmental trends in the course of pregnancy. Sideway bendings increased rapidly between 24 and 30 days and declined hereafter. General movements and fetal breathing increased during midpregnancy and declined towards parturition. Conversely, fetal rest was observed for approximately 60% of time at midgestation and a marked increase was found towards parturition. There were no significant differences in developmental trend of the various movement patterns between individual fetuses. Fetal motility in the guinea pig followed a specific temporal pattern, like in the human, but at a different time scale. The present quantitative data will enable functional investigations into the role of the neuromuscular system. They may also facilitate studies on the effect of environmental influences, such as stress, drugs, toxic substances, and food conditions, on fetal neurobehavioural development in this species. PMID- 19560477 TI - Executive cognitive functions and impulsivity as correlates of risk taking and problem behavior in preadolescents. AB - Initiation of drug use and other risky behavior in preadolescence is associated with poor developmental outcomes. In this research, we examine models that ascribe the trajectory to (a) weak executive cognitive function (ECF), (b) early manifestation of externalizing problems, or (c) heightened levels of trait impulsivity. We test the explanatory power of these factors in a structural equation model with a community sample of 387 preadolescents ages 10-12 years. Participants were tested with a computerized battery of tasks to assess three facets of ECF (working memory, cognitive control, and reward processing) as well as with an audio assisted computerized self-interview to obtain reports of impulsivity and risk behaviors (use of cigarettes and alcohol as well as engaging in fighting and gambling for money) and a self-administered questionnaire to assess externalizing and internalizing problems. The best fitting model explained both early risk taking and externalizing symptoms as the result of individual differences in impulsivity. Although no ECF was directly related to risk taking, working memory and one measure of reward processing performance (reversal learning) were inversely related to impulsivity. The results are discussed in regard to theories of early risk taking with particular focus on the potential relation between ECF and impulsive behavior tendencies and the implications for early intervention to prevent the dysfunctional trajectory associated with early risk behavior. PMID- 19560479 TI - Anxiety-induced cognitive bias in non-human animals. AB - As in humans, 'cognitive biases' in the way in which animals judge ambiguous stimuli may be influenced by emotional state and hence a valuable new indicator of animal emotion. There is increasing evidence that animals experiencing different emotional states following exposure to long-term environmental manipulations show contrasting biases in their judgement of ambiguous stimuli. However, the specific type of induced emotional state is usually unknown. We investigated whether a short-term manipulation of emotional state has a similar effect on cognitive bias, using changes in light intensity; a treatment specifically related to anxiety-induction. Twenty-four male rats were trained to discriminate between two different locations, in either high ('H') or low ('L') light levels. One location was rewarded with palatable food and the other with aversive food. Once the rats had shown spatial discrimination, by running significantly faster to the rewarded location, they were tested with three ambiguous locations intermediate between the rewarded and aversive locations, and their latency to approach each location recorded. Half the rats were tested in the same light levels as during training, the remainder were switched. Rats switched from high to low light levels (putatively the least negative emotional manipulation) ran significantly faster to all three ambiguous probes than those rats switched from low to high light levels (putatively the most negative manipulation). This suggests that the judgement bias technique might be useful as an indicator of short-term changes in anxiety for non-human animals. PMID- 19560480 TI - Social stress enhances IL-1beta and TNF-alpha production by Porphyromonas gingivalis lipopolysaccharide-stimulated CD11b+ cells. AB - Psychological stress is associated with an increased expression of markers of peripheral inflammation, and there is a growing literature describing a link between periodontal pathogens and systemic inflammation. The hypothesis of the present work is that exposing mice to the social stressor, called social disruption (SDR), would enhance the inflammatory response to lipopolysaccharide (LPS) derived from the oral pathogen, Porphyromonas gingivalis. Mice were exposed to SDR for 2h per day on 6 consecutive days. On the morning following the last cycle of SDR, mice were tested for anxiety-like behavior in the open field test and novel object test. The mice were sacrificed the following day and their spleens harvested. Spleen cells were stimulated with LPS derived from P. gingivalis in the absence or presence of increasing doses of corticosterone. Social disruption resulted in anxiety-like behavior, and the production of IL 1beta and TNF-alpha was significantly higher in spleen cells from mice exposed to SDR in comparison to levels from non-stressed control mice. In addition, the viability of spleen cells from mice exposed to SDR was significantly greater than the viability of cells from non-stressed control mice, even in the presence of high doses of corticosterone. The use of cultures enriched for CD11b+ cells indicated that the stressor was affecting the activity of splenic myeloid cells. This study demonstrates that social stress enhances the inflammatory response to an oral pathogen and could provide a critical clue in the reported associations between stress, inflammation, and oral pathogens. PMID- 19560481 TI - Neurobehavioral impairments, generation of oxidative stress and release of pro apoptotic factors after chronic exposure to sulphur mustard in mouse brain. AB - Recent global events have focused attention on the potential threat of international and domestic chemical terrorism, as well as the possibility of chemical warfare proliferation. Sulphur mustard (SM) is one of the potent chemical warfare agents (CWA), which initiates a cascade of events that converge on the redox mechanisms common to brain injury. The present study was designed to examine the effects of chronic SM exposure on neurobehavioral impairments, mitochondrial oxidative stress in male Swiss Albino mice and its role in inducing apoptotic neuronal cell death. The animals were divided into four groups (control, low, medium and high dose) of 5 animals each. Exposure to SM was given percutaneously daily for 12 weeks. The results demonstrated impairment in neurobehavioral indices viz. rota rod, passive avoidance and water maze tests in a dose dependent manner. There was a significant increase in lipid peroxidation and protein carbonyl content whereas, decrease in the activity of manganese superoxide dismutase (MnSOD), glutathione reductase and glutathione peroxidase suggesting impaired antioxidant defense system. Immunoblotting of cytochrome c, Bcl-2, Bax and activation of caspase-3 suggest induction of apoptosis in a dose dependent manner. Finally, increased p53 expression suggests that it may target the mitochondrial pathway for inducing apoptosis in response to DNA damage signals. In conclusion, chronic SM exposure may have the potential to generate oxidative stress which may trigger the release of cytochrome c as well as caspase 3 activation in neurons leading to cell death by apoptosis in a dose dependent manner which may in the end be responsible for the disruption of cognitive functions in mice. PMID- 19560482 TI - Cysteine-based regulation of the CUL3 adaptor protein Keap1. AB - Nrf2 (NF-E2-related factor 2) is a master transcription factor containing a powerful acidic transcriptional activation domain. Nrf2-dependent gene expression impacts cancer chemoprevention strategies, inflammatory responses, and progression of neurodegenerative diseases. Under basal conditions, association of Nrf2 with the CUL3 adaptor protein Keap1 results in the rapid Nrf2 ubiquitylation and proteasome-dependent degradation. Inhibition of Keap1 function blocks ubiquitylation of Nrf2, allowing newly synthesized Nrf2 to translocate into the nucleus, bind to ARE sites and direct target gene expression. Site-directed mutagenesis experiments coupled with proteomic analysis support a model in which Keap1 contains at least 2 distinct cysteine motifs. The first is located at Cys 151 in the BTB domain. The second is located in the intervening domain and centers around Cys 273 and 288. Adduction or oxidation at Cys151 has been shown to produce a conformational change in Keap1 that results in dissociation of Keap1 from CUL3, thereby inhibiting Nrf2 ubiquitylation. Thus, adduction captures specific chemical information and translates it into biochemical information via changes in structural conformation. PMID- 19560484 TI - Saxitoxins accumulation by freshwater tilapia (Oreochromis niloticus) for human consumption. AB - The accumulation of saxitoxins (STXs) in fish from freshwater aquaculture was investigated for the first time in the present study. Cyanotoxins have been monitored in liver and muscle samples of Oreochromis niloticus by chromatographic methods, both before and after the depuration process. The results show that tilapia can accumulate STXs. Our findings suggest that depuration with clean water is an alternative process to eliminate STXs from fish and, therefore, improve the safety of tilapia for consumers. PMID- 19560486 TI - Anthrax toxins: a weapon to systematically dismantle the host immune defenses. AB - Successful colonization of the host by bacterial pathogens relies on their capacity to evade the complex and powerful defenses opposed by the host immune system, at least in the initial phases of infection. The two toxins of Bacillus anthracis, lethal toxin and edema toxin, appear to have been shaped by evolution to assist the microorganism in this crucial function, in addition to act as general toxins acting on almost all cell types. Edema toxin causes a consistent elevation of cAMP, an important second messenger the production of which is normally strictly controlled in mammalian cells, whereas lethal toxin cleaves most isoforms of mitogen-activated protein kinase kinases. By disrupting or subverting central modules common to all the principal signaling networks which control immune cell activation, effector function and migration, the anthrax toxins effectively and systematically dismantle both the innate and the adaptive immune defenses of the host. Here, we review the specific effects of the lethal and edema toxins of B. anthracis on the activation and function of phagocytes, dendritic cells and lymphocytes. We also discuss some open issues which should be addressed to gain a comprehensive insight into the complex relationship that B. anthracis establishes with the host. PMID- 19560483 TI - Improving in vitro Sertoli cell/gonocyte co-culture model for assessing male reproductive toxicity: Lessons learned from comparisons of cytotoxicity versus genomic responses to phthalates. AB - Gonocytes exist in the neonatal testis and represent a transient population of male germ-line stem cells. It has been shown that stem cell self-renewal and progeny production is probably controlled by the neighboring differentiated cells and extracellular matrix (ECM) in vivo known as niches. Recently, we developed an in vitro three-dimensional (3D) Sertoli cell/gonocyte co-culture (SGC) model with ECM overlay, which creates an in vivo-like niche and supports germ-line stem cell functioning within a 3D environment. In this study, we applied morphological and cytotoxicity evaluations, as well as microarray-based gene expression to examine the effects of different phthalate esters (PE) on this model. Known in vivo male developmentally toxic PEs (DTPE) and developmentally non-toxic PEs (DNTPE) were evaluated. We observed that DTPE induced significantly greater dose-dependent morphological changes, a decrease in cell viability and an increase in cytotoxicity compared to those treated with DNTPE. Moreover, the gene expression was more greatly altered by DTPE than by DNTPE and non-supervised cluster analysis allowed the discrimination of DTPE from the DNTPE. Our systems-based GO Quant analysis showed significant alterations in the gene pathways involved in cell cycle, phosphate transport and apoptosis regulation with DTPE but not with DNTPE treatment. Disruptions of steroidogenesis related-gene expression such as Star, Cyp19a1, Hsd17b8, and Nr4a3 were observed in the DTPE group, but not in the DNTPE group. In summary, our observation on cell viability, cytotoxicity, and microarray-based gene expression analysis induced by PEs demonstrate that our in vitro 3D-SGC system mimicked in vivo responses for PEs and suggests that the 3D SGC system might be useful in identifying developmental reproductive toxicants. PMID- 19560485 TI - The adenylyl cyclase activity of anthrax edema factor. AB - Bacillus anthracis, the etiologic agent for anthrax, secretes edema factor (EF) to disrupt intracellular signaling pathways. Upon translocation into host cells and association with a calcium sensor, calmodulin (CaM), EF becomes a highly active adenylyl cyclase (AC) that raises the intracellular concentration of cyclic AMP (cAMP). Growing evidence shows that EF plays a key role in anthrax pathogenesis by affecting cellular functions vital for host defense. This strategy is also used by Bordetella pertussis, a bacterium that causes whooping cough. Pertussis bacteria secrete the bifunctional toxin CyaA which raises the intracellular cAMP. Here, we discuss recent advances from structural analyses that reveal the molecular basis of the conserved mechanism of activation and catalysis of EF and CyaA by CaM even though these two toxins use the completely different sequences to bind CaM. Comparison of the biochemical and structural characteristics of these two AC toxins with host ACs reveal that they have diverse strategies of catalytic activation, yet use the same two-metal-ion catalytic mechanism. PMID- 19560487 TI - Evolution of ASABF (Ascaris suum antibacterial factor)-type antimicrobial peptides in nematodes: putative rearrangement of disulfide bonding patterns. AB - ASABF (Ascaris suum antibacterial factor)-type antimicrobial peptides are defensin-like cysteine-rich peptides that are widely distributed in the phylum Nematoda. In known members of ASABF-type antimicrobial peptides, an array consisting of eight cysteine residues is completely conserved. In this study, we report a novel member ASABF-6Cys-alpha, which contains only six cysteine residues, in the pig round worm A. suum. The two cysteine residues deleted in ASABF-6Cys-alpha were not identical to a pair of half-cystine forming a disulfide bridge in ASABF-alpha, suggesting a rearrangement of disulfide bonding patterns. Gene organization and phylogenetic analyses suggested that ASABF-6Cys-alpha was generated from an ancestral ASABF gene after the divergence of Ascaridida from Rhabditida. ASABF-6Cys-alpha transcripts dramatically increased after bacterial injection, suggesting that ASABF-6Cys-alpha may contribute to immunity in nematodes. PMID- 19560488 TI - Gut hormones: implications for the treatment of obesity. AB - Bariatric surgery is the only effective treatment for patients with morbid obesity. This is no solution to the present obesity pandemic however. Currently licensed non-surgical pharmaceuticals are of limited efficacy and alternatives are needed. Harnessing the body's own appetite-regulating signals is a desirable pharmacological strategy. The gastrointestinal tract has a prime role in sensing and signalling food intake to the brain. Gut hormones are key mediators of this information, including: peptide YY (PYY), pancreatic polypeptide (PP), glucagon like peptide 1 (GLP-1), oxyntomodulin (OXM), ghrelin, amylin and cholecystokinin (CCK). This review summarises the latest knowledge regarding the physiological and pathophysiological role of gut hormones in regulating our food intake and how this knowledge could guide, or has guided, the development of weight-loss drugs. Up-to-date outcomes of clinical trials are evaluated and directions for the future suggested. PMID- 19560490 TI - Flavopiridol reduces the impedance of neural prostheses in vivo without affecting recording quality. AB - We hypothesized that re-entry into the cell cycle may be associated with reactive gliosis surrounding neural prostheses, and that administration of a cell cycle inhibitor (flavopiridol) at the time of surgery would reduce this effect. We investigated the effects of flavopiridol on recording quality and impedance over a 28-day time period and conducted histology at 3 and 28 days post-implantation. Flavopiridol reduced the expression of a cell cycle protein (cyclin D1) in microglia surrounding probes at the 3-day time point. Impedance at 1 kHz was decreased by drug administration across the study period compared to vehicle controls. Correlations between recording (SNR, units) and impedance metrics revealed a small, but statistically significant, inverse relationship between these variables. However, the relationship between impedance and recording quality was not sufficiently strong for flavopiridol to result in an improvement in SNR or the number of units detected. Our data indicate that flavopiridol is an effective, easily administered treatment for reducing impedance in vivo, potentially through inhibiting microglial encapsulation of implanted devices. This strategy may be useful in stimulation applications, where reduced impedance is desirable for achieving activation thresholds and prolonging the lifetime of the implanted power supply. While improvements in recording quality were not observed, a combination of flavopiridol with a second strategy which enhances neuronal signal detection may enhance these results in future studies. PMID- 19560491 TI - The impact of synaptic conductance on action potential waveform: evoking realistic action potentials with a simulated synaptic conductance. AB - Most current clamp studies trigger action potentials (APs) by step current injection through the recording electrode and assume that the resulting APs are essentially identical to those triggered by orthodromic synaptic inputs. However this assumption is not always valid, particularly when the synaptic conductance is of large magnitude and of close proximity to the axon initial segment. We addressed this question of similarity using the Calyx of Held/MNTB synapse; we compared APs evoked by long duration step current injections, short step current injections and orthodromic synaptic stimuli. Neither injected current protocol evoked APs that matched the evoked orthodromic AP waveform, showing differences in AP height, half-width and after-hyperpolarization. We postulated that this 'error' could arise from changes in the instantaneous conductance during the combined synaptic and AP waveforms, since the driving forces for the respective ionic currents are integrating and continually evolving over this time-course. We demonstrate that a simple Ohm's law manipulation of the EPSC waveform, which accounts for the evolving driving force on the synaptic conductance during the AP, produces waveforms that closely mimic those generated by physiological synaptic stimulation. This stimulation paradigm allows supra-threshold physiological stimulation (single stimuli or trains) without the variability caused by quantal fluctuation in transmitter release, and can be implemented without a specialised dynamic clamp system. Combined with pharmacological tools this method provides a reliable means to assess the physiological roles of postsynaptic ion channels without confounding affects from the presynaptic input. PMID- 19560489 TI - Laser-evoked synaptic transmission in cultured hippocampal neurons expressing channelrhodopsin-2 delivered by adeno-associated virus. AB - We present a method for studying synaptic transmission in mass cultures of dissociated hippocampal neurons based on patch clamp recording combined with laser stimulation of neurons expressing channelrhodopsin-2 (ChR2). Our goal was to use the high spatial resolution of laser illumination to come as close as possible to the ideal of identifying monosynaptically coupled pairs of neurons, which is conventionally done using microisland rather than mass cultures. Using recombinant adeno-associated virus (rAAV) to deliver the ChR2 gene, we focused on the time period between 14 and 20 days in vitro, during which expression levels are high, and spontaneous bursting activity has not yet started. Stimulation by wide-field illumination is sufficient to make the majority of ChR2-expressing neurons spike. Stimulation with a laser spot at least 10 microm in diameter also produces action potentials, but in a reduced fraction of neurons. We studied synaptic transmission by voltage-clamping a neuron with low expression of ChR2 and scanning a 40 microm laser spot at surrounding locations. Responses were observed to stimulation at a subset of locations in the culture, indicating spatial localization of stimulation. Pharmacological means were used to identify responses that were synaptic. Many responses were of smaller amplitude than those typically found in microisland cultures. We were unable to find an entirely reliable criterion for distinguishing between monosynaptic and polysynaptic responses. However, we propose that postsynaptic currents with small amplitudes, simple shapes, and latencies not much greater than 8 ms are reasonable candidates for monosynaptic interactions. PMID- 19560492 TI - Autophagy protein 8 mediating autophagosome in encysting Acanthamoeba. AB - Autophagy is an evolutionally conserved protein degradation pathway in eukaryotes. It plays essential roles during starvation, cellular differentiation, cell death, and aging by eliminating unwanted or unnecessary organelles and recycling the components for reuse. ATG8, a member of a novel ubiquitin-like protein family, is an essential component of the autophagic machinery. The present study identified and characterized autophagy protein 8 in Acanthamoeba castellanii an amphizoic amoeba causing granulomatous amoebic encephalitis and amoebic keratitis in humans. Real-time polymerase chain reaction demonstrated that the A. castellanii Atg8 (AcAtg8) gene encoding a 118 amino acid protein was highly expressed during encystation. Fluorescence microscopic analysis following transient transfection of enhanced green fluorescent protein-AcAtg8 revealed small or large vacuolar fluorescent structures in an encysting amoeba. The Atg8 fluorescent structures on the membrane were identified as autophagosomes by co localization analysis with LysoTracker. Chemically synthesized small interfering RNA against AcAtg8 reduced the encystation efficiency and inhibited autophagosome formation in Acanthamoeba. PMID- 19560493 TI - Maternal isobutyl-paraben exposure alters anxiety and passive avoidance test performance in adult male rats. AB - Isobutyl-paraben (IBP), one of the most widely used preservatives, exhibits estrogenic activity. In this study, we analyzed the effects of maternal IBP treatment on the emotional behavior and learning performance in mature offspring. Pregnant female Sprague-Dawley rats were treated with IBP via a subcutaneous Silastic capsule. Consequently, the offspring were exposed to IBP during gestation through the placentae, and before weaning through the milk. Male and female offspring were tested for emotional behavior in an open field and in an elevated plus maze at five and six weeks old, respectively. IBP-exposed male (but not female) rats spent less time in the open arms of the elevated plus maze. At 11 weeks old, all females were gonadectomized and treated chronically with 17beta estradiol or cholesterol by Silastic capsules; all males were kept intact. They were tested for learning performance in a passive avoidance test and a Morris water maze. IBP exposure impaired the performance of males in the passive avoidance test. These findings suggest that male rats are more affected by early exposure to IBP than female rats. IBP affects their adult behavior including anxiety and learning abilities. PMID- 19560494 TI - Metabolic flux analysis of a phenol producing mutant of Pseudomonas putida S12: verification and complementation of hypotheses derived from transcriptomics. AB - The physiological effects of genetic and transcriptional changes observed in a phenol producing mutant of the solvent-tolerant Pseudomonas putida S12 were assessed with metabolic flux analysis. The upregulation of a malate/lactate dehydrogenase encoding gene could be connected to a flux increase from malate to oxaloacetate. A mutation in the pykA gene decreased in vitro pyruvate kinase activity, which is consistent with a lower flux from phosphoenolpyruvate to pyruvate. Changes in the oprB-1, gntP and gnuK genes, encoding a glucose selective porin, gluconokinase and a gluconate transporter respectively, altered the substrate uptake profile. Metabolic flux analysis furthermore revealed cellular events not predicted by the transcriptome analysis. Gluconeogenic formation of glucose-6-phosphate from triose-3-phosphate was abolished, in favour of increased phosphoenolpyruvate production. An increased pentose phosphate pathway flux resulted in higher erythrose-4-phosphate production. Thus, the availability of these two central phenol precursors was improved. Furthermore, metabolic fluxes were redistributed such that the overall TCA cycle flux was unaffected and energy production increased. Engineering P. putida S12 for phenol production has yielded a strain that channels carbon fluxes to previously unfavourable routes to reconcile the drain on metabolites required for phenol production, while maintaining basal flux levels through central carbon metabolism. PMID- 19560495 TI - Crabtree-negative characteristics of recombinant xylose-utilizing Saccharomyces cerevisiae. AB - For recombinant xylose-utilizing Saccharomyces cerevisiae, ethanol yield and productivity is substantially lower on xylose than on glucose. In contrast to glucose, xylose is a novel substrate for S. cerevisiae and it is not known how this substrate is recognized on a molecular level. Failure to activate appropriate genes during xylose-utilization has the potential to result in sub optimal metabolism and decreased substrate uptake. Certain differences in fermentative performance between the two substrates have thus been ascribed to variations in regulatory response. In this study differences in substrate utilization of glucose and xylose was analyzed in the recombinant S. cerevisiae strain TMB3400. Continuous cultures were performed with glucose and xylose under carbon- and nitrogen-limited conditions. Whereas biomass yield and substrate uptake rate were similar during carbon-limited conditions, the metabolic profile was highly substrate dependent under nitrogen-limited conditions. While glycerol production occurred in both cases, ethanol production was only observed for glucose cultures. Addition of acetate and 2-deoxyglucose pulses to a xylose limited culture was able to stimulate transient overflow metabolism and ethanol production. Application of glucose pulses enhanced xylose uptake rate under restricted co-substrate concentrations. Results are discussed in relation to regulation of sugar metabolism in Crabtree-positive and -negative yeast. PMID- 19560496 TI - Characterization of short-lived intermediates produced during replication of baculovirus DNA. AB - In this report the short-lived DNA replication intermediates produced in both uninfected and Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infected Spodoptera frugiperda cells were characterized. The methods used included pulse-labeling of DNA in permiabilized cells, treatment of nascent DNA with Mung bean nuclease, and electrophoresis in neutral and alkaline agarose gels. In contrast to uninfected cells that produced a population of small DNA fragments of about 200bp, a population of heterogeneous fragments of up to 5kb with an average size of 1-2kb derived randomly from the virus genome was identified as the short-lived intermediates produced during AcMNPV replication. The intermediates likely include Okazaki fragments derived from the lagging strands in viral replication forks as well as fragments produced during the recombination-dependent replication. PMID- 19560497 TI - Hydrotropic oligomer-conjugated glycol chitosan as a carrier of paclitaxel: synthesis, characterization, and in vivo biodistribution. AB - Development of successful formulations for poorly water-soluble drugs remains a longstanding critical and challenging issue in cancer therapy. As a potential drug carrier of paclitaxel, hydrotropic oligomer-glycol chitosan (HO-GC) was synthesized by chemical conjugation of the N,N-diethylnicotinamide-based oligomer, uniquely designed for enhancing the aqueous solubility of paclitaxel, to the backbone of glycol chitosan. Owing to its amphiphilicity, the conjugate formed self-assembled nanoparticles with a mean diameter of 313+/-13nm in a phosphate-buffered saline (PBS, pH 7.4 at 37 degrees C). HO-GC nanoparticles maintained their structure for up to 50days in PBS. They could encapsulate a high quantity (20wt.%) of paclitaxel (PTX) with a maximum drug-loading efficiency of 97%, due to the presence of hydrotropic inner cores. When HO-GC-PTX particles were exposed to the 0.1M sodium salicylate solution in PBS (pH 7.4), PTX was released from nanoparticles in a sustained manner. From the cytotoxicity test, it was confirmed that HO-GC-PTX nanoparticles showed lower cytotoxicity than free PTX formulation in 50%/50% Cremophor EL/ethanol mixture. The optical imaging results indicated that near-infrared fluorescence dye (Cy5.5)-labeled HO-GC-PTX showed an excellent tumor specificity in SCC7 tumor-bearing mice, due to the enhanced permeation and retention effect. Overall, HO-GC-PTX nanoparticles might be a promising carrier for PTX delivery in cancer therapy. PMID- 19560498 TI - Effects of Manduca sexta allatostatin and an analog on the pea aphid Acyrthosiphon pisum (Hemiptera: Aphididae) and degradation by enzymes from the aphid gut. AB - The C-type allatostatin, Manduca sexta allatostatin (Manse-AS) and the analog delta R(3)delta R(5)Manse-AS, where R residues were replaced by their d-isomers, were tested for oral toxicity against the pea aphid Acyrthosiphon pisum (Harris) by incorporation into an artificial diet. Both peptides had significant dose dependent feeding suppression effects, resulting in mortality, reduced growth and fecundity compared with control insects. The delta R(3)delta R(5)Manse-AS analog had an estimated LC(50) of 0.18 microg/microl diet, and was more potent than Manse-AS. At a dose of 0.35 microg delta R(3)delta R(5)Manse-AS/microl diet, 98% of aphids were dead within 3 days, at a rate similar to those aphids that had been starved (no diet controls). On comparison, it required 13 days and three times the dose of Manse-AS fed to aphids to attain 96% mortality. It is possible that the feeding suppression effects of Manse-AS on aphids are due to the inhibition of gut motility. The estimated half-life of Manse-AS when incubated with a gut extract from A. pisum was 54 min. Degradation was most likely due to cathepsin L cysteine and/or trypsin-like proteases, by an unidentified glutamine specific protease and by a carboxypeptidase-like enzyme. The d-isomers of R in the Manse-AS analog appeared to prevent hydrolysis by cathepsin L cysteine and trypsin-like enzymes, and enhance its half-life (145 min). However delta R(3)delta R(5)Manse-AS was cleaved by enzymes with carboxypeptidase-like and chymotrypsin-like activity. The increased stability of the Manse-AS analog may explain its enhanced feeding suppression effects when continually fed to aphids, and demonstrates the potential use of Manse-AS in a strategy to control aphid pests. PMID- 19560499 TI - In vitro antimicrobial activity of alpha-melanocyte stimulating hormone against major human pathogen Staphylococcus aureus. AB - Alpha-melanocyte stimulating hormone (alpha-MSH) is an endogenous anti inflammatory peptide reported to possess antimicrobial properties, however their role as antibacterial peptides is yet to be established. In the present study, we examined in vitro antibacterial activity of alpha-MSH against S. aureus strain ISP479C and several methicillin-sensitive (MSSA) and methicillin-resistant (MRSA) S. aureus strains. Antibacterial activity was examined by varying several parameters, viz., bacterial cell densities, growth phase, pH, salt concentration, and temperature. Antibacterial activity was also examined in complex biomatrices of rat whole blood, plasma and serum as well as in biofilm form of bacteria. Our results showed that alpha-MSH possessed significant and rapid antibacterial activity against all the studied strains including MRSA (84% strains were killed on exposure to 12 microM of alpha-MSH for 2h). pH change from 7.4 to 4 increased alpha-MSH staphylocidal activity against ISP479C by 21%. Antibacterial activity of alpha-MSH was dependent on bacterial cell density and independent of growth phase. Moreover, antimicrobial activity was retained when alpha-MSH was placed into whole blood, plasma, and serum. Most importantly, alpha-MSH exhibited antibacterial activity against staphylococcal biofilms. Multiple membrane permeabilization assays suggested that membrane damage was, at least in part, a major mechanism of staphylocidal activity of alpha-MSH. Collectively the above findings suggest that alpha-MSH could be a promising candidate of a novel class of antimicrobial agents. PMID- 19560501 TI - Plasma nociceptin/orphanin FQ levels rise after spontaneous episodes of angina, but not during induced myocardial ischemia. AB - The aim of our study was to evaluate the effects of repeated episodes of angina and induced myocardial ischemia on plasma nociceptin/orphanin FQ (N/OFQ) levels. Patients with unstable angina (23 with new onset severe angina or accelerated angina and 18 with subacute angina at rest) who had had repeated spontaneous episodes of chest pain in the last week before the study underwent myocardial perfusion single-photon emission computed tomography using adenosine infusion. Twenty subjects without clinical symptoms of angina matched for age, sex and cardiac risk factors served as a control group. N/OFQ levels were significantly (P<0.01) higher in the patients (15.2+/-2.1 pg/ml) than in the control group (8.5+/-2.6 pg/ml). Blood pressure and heart rate did not significantly differ. All patients showed transient adenosine infusion myocardial ischemia that did not induce chest pain or significantly modify plasma N/OFQ levels or hemodynamic parameters. Our findings show that unstable angina is associated with a significant increase in circulating N/OFQ levels unrelated to intervening transient myocardial ischemia or hemodynamic changes. This increase is probably related to the chest pain repeatedly occurring in the course of coronary artery disease, but absent during transient adenosine-induced myocardial ischemia. PMID- 19560500 TI - Glucagon-like peptide-1 protects mesenteric endothelium from injury during inflammation. AB - Glucagon-like peptide-1 (GLP-1) is a proglucagon-derived hormone with cellular protective actions. We hypothesized that GLP-1 would protect the endothelium from injury during inflammation. Our aims were to determine the: (1) effect of GLP-1 on basal microvascular permeability, (2) effect of GLP-1 on increased microvascular permeability induced by lipopolysaccaride (LPS), (3) involvement of the GLP-1 receptor in GLP-1 activity, and (4) involvement of the cAMP/PKA pathway in GLP-1 activity. Microvascular permeability (L(p)) of rat mesenteric post capillary venules was measured in vivo. First, the effect of GLP-1 on basal L(p) was measured. Second, after systemic LPS injection, L(p) was measured after subsequent perfusion with GLP-1. Thirdly, L(p) was measured after LPS injection and perfusion with GLP-1+GLP-1 receptor antagonist. Lastly, L(p) was measured after LPS injection and perfusion with GLP-1+inhibitors of the cAMP/PKA pathway. Results are presented as mean area under the curve (AUC)+/-SEM. GLP-1 had no effect on L(p) (AUC: baseline=27+/-1.4, GLP-1=1+/-0.4, p=0.08). LPS increased L(p) two-fold (AUC: LPS=54+/-1.7, p<0.0001). GLP-1 reduced the LPS increase in L(p) by 75% (AUC: LPS+GLP-1=34+/-1.5, p<0.0001). GLP-1 antagonism reduced the effects of GLP-1 by 60% (AUC: LPS+GLP-1+antagonist=46+/-2.0, p<0.001). The cAMP synthesis inhibitor reduced the effects of GLP-1 by 60% (AUC: LPS+GLP-1+cAMP inhibitor=46+/-1.5, p<0.0001). The PKA inhibitor reduced the effects of GLP-1 by 100% (AUC: LPS+GLP-1+PKA inhibitor=56+/-1.5, p<0.0001). GLP-1 attenuates the increase in microvascular permeability induced by LPS. GLP-1 may protect the endothelium during inflammation, thus decreasing third-space fluid loss. PMID- 19560502 TI - Association of obestatin with blood pressure in the third trimesters of pregnancy. AB - Obestatin is a recently discovered 23-amino acid peptide encoded by the same gene that encodes ghrelin. It has been reported that there is a significant negative correlation between the plasma ghrelin concentration and systemic blood pressure in patients with pregnancy-induced hypertension. We investigated the plasma concentration of obestatin in 18 non-pregnant women, 18 normal pregnant women, and 15 patients with pregnancy-induced hypertension. The plasma concentrations of obestatin in these 3 groups of women were 63.4+/-9.5pg/ml, 38.1+/-6.3pg/ml, and 46.0+/-9.3pg/ml, respectively. In non-pregnant women, there was no correlation between the plasma obestatin concentration and the mean arterial pressure. However, there was a positive correlation between the plasma obestatin concentration and the mean arterial pressure in normal pregnant women and pregnant women with pregnancy-induced hypertension. These results suggest that obestatin may have some potential role in the regulation of blood pressure in normal pregnant women and women with pregnancy-induced hypertension. PMID- 19560503 TI - Axodendritic fibres of mouse cerebellar granule neurons exhibit a diversity of functional P2X receptors. AB - Distribution and functional expression of P2X receptors were analyzed in mouse cerebellum axodendritic fibres, using different experimental approaches such as RT-PCR, western blot, immunochemistry, microfluorimetric experiments and exocytotic studies. RT-PCR and western blot demonstrated the presence of P2X1-4,7 subunits in both whole cerebellum and mouse cerebellar granule cultured neurons. Immunochemistry analysis of tissular and cellular location of P2X1-4,7 receptors confirmed their presence and unequal distribution between somas and axodendritic prolongations. Microfluorimetric experiments using a variety of modulators of the P2X subunits revealed the presence of different functional P2X receptors in the axodendritic fibres. The use of the synthetic agonist alpha,beta-meATP and the antagonist Ip(5)I revealed the activation of functional P2X1 and P2X3 receptors. Responses mediated by P2X1 subunits were also confirmed by using ZnSO(4). Activation of functional P2X4 receptors is observed when stimulated in the presence of ivermectin. Exocytotic studies confirmed the role of most P2X subunits in the activation of neurotransmitter release in axodendritic fibres from mouse cerebellar granule neurons. PMID- 19560504 TI - Oxidative stress promotes JNK-dependent amyloidogenic processing of normally expressed human APP by differential modification of alpha-, beta- and gamma secretase expression. AB - The pathogenesis of Alzheimer disease (AD) is complex and is certain to involve diverse etiological factors, but a central role has been strongly suggested for amyloid beta-protein (Abeta), based on genetic, biochemical and neurotoxicological evidence. In contrast with the well-documented effect of genetic mutations in Abeta overproduction, not much is known about the mechanisms involved in sporadic AD (SAD) which account for more than 95% of cases. Extensive data from patients and in vivo animal models indicate that oxidative stress is one of the cardinal factors most frequently associated with this neurodegenerative disease. The aim of the present study was to explore the effect of oxidative stress on the normally expressed wild-type amyloid precursor protein (APP) in human neuroblastoma cells, which represents a more physiological model of neuronal Abeta generation. Since H(2)O(2) is the main source of the highly reactive hydroxyl radical in the brain, and FeCl(2) can stimulate oxidative stress, including the formation of the hydroxyl radical from H(2)O(2), in the present work we studied the effect of these two pro-oxidant molecules on the levels and processing of human APP by alpha-, beta- and gamma-secretase, and the role of the stress-activated kinase c-jun N-terminal kinase (JNK). We provide evidence for a dual modulation of amyloid precursor protein metabolism in differentiated human neuroblastoma cells related with a down-regulation of alpha secretase and up-regulation of gamma-secretase, and particularly of beta secretase and also a JNK depending Abeta generation. PMID- 19560505 TI - Alterations in excitotoxicity and prostaglandin metabolism in a transgenic mouse model of Alzheimer's disease. AB - To address the potential impact of presenilin mutations on the prostaglandin metabolism in a neurodegenerative model of glutamatergic excitotoxicity, we injected kainic acid intraperitoneally (30mg/kg body weight) into mice over expressing the human N141I mutation of presenilin-2, which is known to cause an early-onset form of Alzheimer's disease. We compared the seizure activity as well as seizure lethality in 2- and 6-month-old mice, transgenic for the above mentioned point mutation, and their wildtype littermates and found that mice harboring the hN141I mutation showed a relative resistance to excitotoxic treatment. This was associated with a constituitively reduced expression of the cyclooxygenases COX-1 and COX-2 in the hippocampus of N141I presenilin-2 mice and a reduced induction of COX-2 expression post-kainate injection. In the past, clinical trials have suggested that both non-steroidal anti-inflammatory drugs, which impact upon a cell's prostaglandin metabolism, and glutamatergic antagonists might be of benefit to patients suffering from Alzheimer's-type dementias. Yet, the exact mechanism by which these drugs are beneficial remains unclear, although it seems possible that presenilins might be implicated in the process, at least in the case of early-onset forms. The data presented here strongly support the notion of an implication of presenilins in the alterations in the prostaglandin system, which have been observed in Alzheimer's disease and may contribute to the underlying pathogenesis of the disease. PMID- 19560506 TI - The Flinders Sensitive Line of rats, a rat model of depression, has elevated brain glucose utilization when compared to normal rats and the Flinders Resistant Line of rats. AB - The Flinders Sensitive Line (FSL) of rats, developed as a line of rats with behavioral supersensitivity to diisopropyl fluorophosphates, a cholinesterase inhibitor, has been used as a rat model of depression. This line has many behavioral and neurochemical similarities to human depression. Because brain regional glucose utilization has been extensively studied in patients with depression with a rather heterogeneous outcome, it became necessary to evaluate this parameter in a well defined and homogeneous animal model. In the present study, the regional glucose utilization measured in thirty-six distinct brain regions and the pineal body was determined in FSL rats and the values were compared to those found in normal Sprague-Dawley (SPD) rats and the Flinders Resistant Line of rats (FRL: animals which have a high resistance to the above noted cholinesterase inhibitor). The measurements were performed by autoradiography with 2-[(14)C]deoxyglucose (2-DG) as the tracer. SPD and FRL groups consisted of six and FSL had 8 rats. The rats were sacrificed 45min following the start of the tracer injection. The results showed that the global, and, in many brain areas, regional, glucose utilization is higher in the FSL rats than in the control SPD rats. The synthesis in the FRL rats was very similar to that of the SPD rats, but it was different in many brain regions from the FSL rats. Because the FSL rats have many neurochemical and behavioral differences, in particular, those parameters having lower values, the results presented here suggest that the biochemical processes in the brain of FSL rats occur with lower efficiency than in either the normal SPD rats or FRL rats. PMID- 19560507 TI - Interaction of serotonin-related genes affects short-term antidepressant response in major depressive disorder. AB - BACKGROUND: Four serotonin-related genes including guanine nucleotide binding protein beta polypeptide 3 (GNB3), 5-hydroxytryptamine receptor 1A (HTR1A; serotonin receptor 1A), 5-hydroxytryptamine receptor 2A (HTR2A; serotonin receptor 2A), and solute carrier family 6 member 4 (SLC6A4; serotonin neurotransmitter transporter) have been suggested to be candidate genes for influencing antidepressant treatment outcome. The aim of this study was to explore whether interaction among these genes could contribute to the pharmacogenomics of short-term antidepressant response in a Taiwanese population with major depressive disorder (MDD). METHODS: Included in this study were 101 MDD patients who were treated with antidepressants, 35 of whom were rapid responders and 66 non-responders after 2weeks of treatment. We genotyped four single nucleotide polymorphisms (SNPs), including GNB3 rs5443 (C825T), HTR1A rs6295 (C-1019G), HTR2A rs6311 (T102C), and SLC6A4 rs25533, and employed the generalized multifactor dimensionality reduction (GMDR) method to investigate gene-gene interactions. RESULTS: Single-locus analyses showed the GNB3 rs5443 polymorphism to be associated with short-term antidepressant treatment outcome (P value=0.029). We did not correct for multiple testing in these multiple exploratory analyses. Finally, the GMDR approach identified a significant gene gene interaction (P-value=0.025) involving GNB3 and HTR2A, as well as a significant 3-locus model (P-value=0.015) among GNB3, HTR2A, and SLC6A4. CONCLUSIONS: These results support the hypothesis that GNB3, HTR2A, and SLC6A4 may play a role in the outcome of short-term antidepressant treatment for MDD in an interactive manner. Future research with independent replication using large sample sizes is needed to confirm the functions of the candidate genes identified in this study as being involved in short-term antidepressant treatment response. PMID- 19560508 TI - Pulmonary exposure to diesel exhaust particles promotes cerebral microvessel thrombosis: protective effect of a cysteine prodrug l-2-oxothiazolidine-4 carboxylic acid. AB - Inhaled particulate matter is associated with increased cerebro- and cardiovascular events. However, the systemic mechanisms underlying these effects remain unclear. In the present study, we investigated the mechanisms underlying the relationship between airway and systemic inflammation and pial cerebral venular thrombosis, 24h after intratracheal (i.t.) instillation of diesel exhaust particles (DEP; 15 or 30 microg/mouse) or saline (control). Doses of 15 and 30 microg/mouse induced a dose-dependent macrophage and neutrophil influx into the bronchoalveolar lavage (BAL) fluid with elevation of total proteins and Trolox equivalent antioxidant capacity (TEAC), but without IL-6 release. Similarly, in plasma, IL-6 concentrations did not increase but the TEAC was significantly and dose-dependently decreased. The number of platelets and the tail bleeding time were both significantly reduced after exposure to DEP (30 microg). Interestingly, the same dose showed platelet proaggregatory effect in mouse pial cerebral venules. Pretreatment with the cysteine prodrug l-2-oxothiazolidine-4-carboxylic acid (OTC, 80 mg/kg) 24 and 1h before i.t. DEP (30 microg), abolished the DEP induced macrophage and neutrophil influx, and the increase of TEAC in BAL. Lung histopathology confirmed the protective effect of OTC on DEP-induced lung inflammation. OTC also reversed the decrease of TEAC concentrations in plasma, the shortening of the bleeding time, and the thrombotic effect of DEP in pial cerebral venules. We conclude that pulmonary exposure to DEP cause oxidative stress responsible, at least partially, for the pulmonary and systemic inflammation and thrombotic events in the pial cerebral microvessels of mice. OTC pretreatment abrogated these effects through its ability to balance oxidant antioxidant status. PMID- 19560509 TI - Assessing neuronal networks: understanding Alzheimer's disease. AB - Findings derived from neuroimaging of the structural and functional organization of the human brain have led to the widely supported hypothesis that neuronal networks of temporally coordinated brain activity across different regional brain structures underpin cognitive function. Failure of integration within a network leads to cognitive dysfunction. The current discussion on Alzheimer's disease (AD) argues that it presents in part a disconnection syndrome. Studies using functional magnetic resonance imaging, positron emission tomography and electroencephalography demonstrate that synchronicity of brain activity is altered in AD and correlates with cognitive deficits. Moreover, recent advances in diffusion tensor imaging have made it possible to track axonal projections across the brain, revealing substantial regional impairment in fiber-tract integrity in AD. Accumulating evidence points towards a network breakdown reflecting disconnection at both the structural and functional system level. The exact relationship among these multiple mechanistic variables and their contribution to cognitive alterations and ultimately decline is yet unknown. Focused research efforts aimed at the integration of both function and structure hold great promise not only in improving our understanding of cognition but also of its characteristic progressive metamorphosis in complex chronic neurodegenerative disorders such as AD. PMID- 19560510 TI - Role and regulation of PI3K in sperm capacitation and the acrosome reaction. AB - Mammalian spermatozoa undergo several signaling and biochemical transformations in the female genital tract, collectively called capacitation. The capacitated spermatozoon binds to the egg zona pellucida, where it undergoes the acrosome reaction (AR), a process enabling it to penetrate and fertilize the egg. Actin polymerization occurs in sperm capacitation and depolymerization prior to the AR. In this review we describe the possible role and regulation of PI3K in sperm capacitation and the acrosome reaction. We claim that PI3K is activated by protein kinase A and suppressed by protein kinase C. Only partial activation of PI3K is seen during the capacitation time, however towards the end of incubation, full activation is observed. Actin polymerization during capacitation is independent on PI3K activity, suggesting that the enzyme is not involved in sperm capacitation. However, the full activation of PI3K towards the end of the capacitation suggests that it might mediate the AR, as indeed was found. PMID- 19560511 TI - Effects of 1-naphthyl acetyl spermine on dendrite formation by cultured cerebellar Purkinje cells. AB - Cerebellar Purkinje cells have the most elaborate dendritic trees among neurons in the brain. To date, the contributions of calcium-permeable AMPA receptors (CP AMPARs) in calcium signaling and dendrite formation of Purkinje cells remain to be elucidated. In the present study, therefore, we examined the effects of 1 naphthyl acetyl spermine (NAS), a blocker of CP-AMPARs, on dendrite formation by cultured Purkinje cells. NAS markedly inhibited elongation and branching of Purkinje cell dendrites. Calcium imaging experiments using caged glutamate demonstrated that NAS inhibits the increase of intracellular calcium concentration in Purkinje cells after glutamate release. These results suggest that calcium signaling mediated through CP-AMPARs plays an important role in Purkinje cell dendrite formation. PMID- 19560512 TI - Mental slowness and executive dysfunctions in patients with metabolic syndrome. AB - Metabolic Syndrome is a cluster of vascular risk factors which has been related to dementia and cognitive decline. The aim of this study was to describe the neuropsychological profile of metabolic syndrome patients. An extensive neuropsychological protocol was administered to 55 patients and 35 controls assessing memory, executive, visuoperceptual and visuoconstructive functions, language and speed of processing. There were differences between groups in speed of processing and some executive functions after controlling for the influences of education and gender. The results suggest that metabolic syndrome may be a prodromal state of vascular cognitive impairment. PMID- 19560514 TI - Evaluation of the vestibulo-ocular reflex using sinusoidal off-vertical axis rotation in patients with acoustic neurinoma. AB - The vestibulo-ocular reflex (VOR) was studied to examine the utility of off vertical axis rotation (OVAR) in the diagnosis of acoustic neurinoma. Subjects were sinusoidally rotated with eyes open in complete darkness at frequencies of 0.4 and 0.8 Hz with a maximum angular velocity of 60 degrees /s at either earth vertical axis rotation (EVAR) or OVAR. Thirteen patients with acoustic neurinomas were investigated. Results showed that VOR gain during OVAR at 0.8 Hz and in a 30 degrees nose-up position in patients with internal auditory canal tumors was significantly less than the gain measured during EVAR. The VOR gain measured from all patients (including those with tumors extending to the cerebellopontine angle) was not significantly different when the patients were subjected to EVAR and OVAR. These observations were possibly due to superior vestibular nerve dysfunction. We concluded that certain stimulating parameters--patient's nose tilted up 30 degrees; sinusoidal OVAR at 0.8 Hz and 60 degrees /s maximum angular head velocity--were useful for evaluating vestibular function in patients suffering from an acoustic neurinoma located within the internal auditory canal. PMID- 19560513 TI - Plasma levels of 24S-hydroxycholesterol reflect brain volumes in patients without objective cognitive impairment but not in those with Alzheimer's disease. AB - OBJECTIVES: Cholesterol has been linked to Alzheimer's disease (AD) and plasma 24S-hydroxycholesterol (24OHC) has been suggested as a surrogate marker for brain cholesterol metabolism. This study investigates the relation of 24OHC as well as markers of extracerebral cholesterol homeostasis (lanosterol, lathosterol, cholesterol, LDL-C, HDL-C and 27-hydroxycholesterol) with brain volumes in memory clinic patients. METHODS: 96 patients (33 with subjective cognitive impairment- SCI; 36 with mild cognitive impairment--MCI; 27 with AD) referred to the Memory Clinic at Karolinska University Hospital, Sweden. Plasma assessments were done by isotope dilution-mass spectrometry. MRI measurements were done using custom-made software BMAP (imaging laboratory, Karolinska Institutet), running on HERMES platform. RESULTS: Ratios of 24-hydroxycholesterol, 27-hydroxycholesterol, lanosterol and lathosterol to cholesterol (R_24OHC, R_27OHC, R_lanosterol and R_lathosterol) were significantly lower in patients with AD. In the whole population, after controlling for age, sex, APOE genotype and statins, R_24OHC was positively related to gray matter (GM) fraction. However, when groups were considered separately, the relation to GM volume, GM and parenchymal fractions was significant in the SCI group only (p<0.05). There was a significant positive association between cholesterol and white matter (WM) volume, WM and parenchymal fractions in patients with AD. CONCLUSIONS: Plasma R_24OHC was lower in patients with AD, but R_24OHC was significantly related to brain volumes in the control group only. One reason may be the previously demonstrated abnormal expression of cholesterol 24S-hydroxylase in astrocytes in AD, which may limit the usefulness of this plasma marker in this specific disease. The findings on cholesterol agree with previous reports of decreasing plasma cholesterol levels in AD patients, suggesting a CNS-mediated effect on extracerebral cholesterol homeostasis. PMID- 19560515 TI - Synchronized diurnal and circadian expressions of four subtypes of melatonin receptor genes in the diencephalon of a puffer fish with lunar-related spawning cycles. AB - Multiple subtypes of melatonin receptors are expressed in neural and peripheral tissues to mediate melatonin actions on the regulation of circadian rhythms in vertebrates. To elucidate molecular basis of "circa" rhythms in the grass puffer Takifugu niphobles, which spawns synchronously with semilunar cycles, tissue distribution of four melatonin receptor subtype mRNAs (Mel(1a) 1.4, Mel(1a) 1.7, Mel(1b), and Mel(1c)) were examined, and diurnal and circadian changes in their absolute amounts were examined in the retina, diencephalon, and optic tectum. Mel(1a) 1.4, Mel(1a) 1.7, and Mel(1b) mRNAs were widely distributed in various brain regions, retina, pituitary, and peripheral tissues, whereas Mel(1c) mRNA was mainly detected in the nervous tissues and pituitary. All subtype genes showed diurnal expressions with one or two peaks during nighttime. When the fish were reared under constant darkness, the retinal expressions of Mel(1a) 1.7, Mel(1b), and Mel(1c) genes were markedly diminished but still showed circadian variations. In contrast, increased and synchronized expressions of the four subtype genes were noticeable with one peak at circadian time 18 in the diencephalon. The circadian expression profiles in the optic tectum were different among the subtypes. The present results suggest that melatonin receptor gene expression is regulated by circadian clock and light, but the effects of light are different among the tissues. The synchronized expressions of the four subtype genes in the diencephalon may be related to the exertion of reproductive rhythmicity in this puffer species. PMID- 19560516 TI - Tyrosine phosphorylation of neuronal nitric oxide synthase (nNOS) during hypoxia in the cerebral cortex of newborn piglets: the role of nitric oxide. AB - The present study aims to investigate the mechanism of activation of nNOS during hypoxia and tests the hypothesis that the hypoxia-induced increased tyrosine phosphorylation of nNOS in the cerebral cortical membranes of newborn piglets is mediated by nNOS-derived nitric oxide (NO). Fifteen newborn piglets were divided into normoxic (Nx, n=5), hypoxic (Hx, n=5) and hypoxic-pretreated with nNOS inhibitor I (Hx-nNOSi) groups. Hypoxia was induced by an FiO(2) of 0.07 for 60 min. nNOS inhibitor I (selectivity>2500 vs endothelial NOS and >500 vs inducible NOS) was administered (0.4 mg/kg, i.v.) 30 min prior to hypoxia. Cortical membranes were isolated and tyrosine phosphorylation of nNOS determined by Western blot. Membrane protein was immunoprecipitated with nNOS antibody, separated on 12% SDS-PAGE and blotted with anti-phosphotyrosine antibody. Protein bands were detected by enhanced chemiluminescence, analyzed by densitometry and expressed as absorbance (OD x mm(2)). Density (OD x mm(2)) of tyrosine phosphorylated nNOS was 51.66+/-14.11 in Nx, 118.39+/-14.17 in Hx (p<0.05 vs Nx) and 45.56+/-10.34 in Hx-nNOSi (p<0.05 vs Hx, p=NS vs Nx). The results demonstrate that pretreatment with nNOS inhibitor prevents the hypoxia-induced increased tyrosine phosphorylation of nNOS. We conclude that the mechanism of hypoxia induced increased tyrosine phosphorylation of nNOS is mediated by nNOS-derived NO. PMID- 19560517 TI - Evidence for transmission disequilibrium at the DAOA gene locus in a schizophrenia family sample. AB - Recently, the DAOA gene locus on chromosome 13q32-q34 has been implicated in the etiology of schizophrenia. We genotyped three single-nucleotide polymorphisms (SNPs: rs778294, rs779293 and rs3918342) in this region in 126 Chinese family trios. In this study, we have identified statistically significant transmission disequilibrium in two markers rs778293 (P=0.01) and rs3918342 (P=0.02), and a highly significant under-transmission between haplotype CAT (P=0.0005) and schizophrenia. The results provide further evidence to support that DAOA gene locus is involved in conferring susceptibility to schizophrenia. PMID- 19560518 TI - Olfactory bulb implantation and methylprednisolone administration in the treatment of spinal cord injury in rats. AB - Estimate the effects of methylprednisolone (MP) administration and olfactory bulb (OB) implantation independently and in combination after a spinal trauma model in Wistar rats, evaluated with BBB scale and CBS with remark of inclined plane test and Tarlov scale. Thirty adult rats were divided into six different groups, evaluated before trauma, one day post-surgery and weekly up to six weeks post lesion. Group A (control); group B (sham) laminectomy without lesion; group C (SCI) lesion only; group D (MP) SCI and MP; group E (OB) SCI and OB implantation; group F (MP/OB) SCI and both therapeutics. Intragroup data at three weeks showed evident significance in groups D, E and F for Tarlov (p=0.001) and BBB (p<0.01); groups C, D, E and F for CBS (p<0.05); and only group D with inclined plane (p<0.05). On the sixth week differences were present in groups C, D, E, and F for Tarlov, BBB and CBS (p<0.001); and C and F for inclined plane (p<0.05). For intergroup analysis any treatment showed differences with Tarlov scale; for BBB and inclined plane, statistical differences were evident in groups E and F; and for CBS only in group F (p<0.05). Real effects of MP are obtained at immediate follow-up, without notorious augmentation after time. OB improvement is achieved only after weeks. None of these therapies used independently achieve a constant and sustained improvement. Combined treatments were more effective and reached higher functional levels for longer periods of time. PMID- 19560519 TI - Association study between the -1021C/T polymorphism of the dopamine-beta hydroxylase gene promoter and personality traits in healthy subjects. AB - Dopamine and norepinephrine are implicated in the characterization of personality traits. Dopamine-beta-hydroxylase (DBH) is the enzyme responsible for conversion of dopamine to norepinephrine, and thus plays an important role in controlling dispositions of these neurotransmitters. Previous studies have shown that the 1021C/T polymorphism of the DBH gene promoter influences plasma DBH activity. Therefore, we examined the association between the -1021C/T DBH polymorphism and personality traits in 627 Japanese healthy volunteers. The DBH genotypes were identified by a PCR-RFLP method, and personality traits were assessed by the Temperament and Character Inventory (TCI). In the two-factor analysis of covariance with the DBH genotype and sex as factors and with age as a covariate, there was no main effect of the DBH genotype on any TCI score, while the interaction between the factors was significant in harm avoidance. In the post hoc analysis, the group with the T allele predictive of lower DBH activity had higher scores of harm avoidance than that without the T allele in females (p=0.006), but not in males. The present study suggests that the -1021C/T DBH polymorphism affects the personality trait of harm avoidance in healthy females. PMID- 19560520 TI - Long-lasting FosB/DeltaFosB immunoreactivity in the rat brain after repeated cat odor exposure. AB - Cat odor elicits defensive responses in rats that are modified by repeated exposure to the odor. We explored the neural correlates of this adaptation using FosB/DeltaFosB immunohistochemistry. Rats were given 3 x60 min exposures to either a worn cat collar (repeated cat odor group) or unworn cat collar (control group). Each exposure was separated by 48h. Rats exposed to cat odor initially showed strong defensive responses (decreased contact, increased retreat, and increased fecal pellets), however these responses had habituated by the final exposure. Rats were perfused 24h or 7 days after the final exposure and the brains were processed for FosB/DeltaFosB protein expression. Up-regulation of FosB/DeltaFosB was observed 24h after repeated cat odor exposure in the nucleus accumbens core, caudate putamen, ventrolateral periaqueductal gray, and four hypothalamic regions: lateral hypothalamus, ventromedial hypothalamus, anterior hypothalamic nucleus, and dorsal premammillary nucleus. FosB/DeltaFosB expression remained elevated 7 days after repeated cat odor exposure in the nucleus accumbens core, caudate putamen and anterior hypothalamic nucleus. These findings provide initial evidence that behavioral changes occurring as a result of repeated predatory stress are associated with long-lasting regionally specific neuroadaptations in the brain. PMID- 19560521 TI - Interaction between neuropeptide Y immunoreactive neurons and galanin immunoreactive neurons in the brain of the masu salmon, Oncorhynchus masou. AB - We investigated the immunohistochemical localization of neuropeptide Y (NPY) and galanin (GAL) in the brain of the masu salmon Oncorhynchus masou in order to clarify the interaction between these neuropeptide hormones in the brain. NPY immunoreactive (ir) cell bodies were observed in the ventral and lateral regions of the ventral telencephalon (Vv and Vl, respectively), and in the dorsolateral midbrain tegmentum. NPY-ir fibers were observed throughout the brain, mainly in the ventral telencephalon, hypothalamus, optic tectum, and midbrain. GAL-ir cell bodies were observed in the nucleus preopticus parvicellularis anterioris (PPa) and the ventral zone of the periventricular hypothalamus (Hv). Both GAL-ir and NPY-ir fibers were observed throughout the brain. Furthermore, we examined the interaction between the NPY neurons and GAL neurons by performing double-staining immunohistochemistry. Some GAL-ir fibers were in close contact with the NPY-ir cell bodies in the Vv and Vl. In addition, some NPY-ir fibers were in close contact with the GAL-ir cell bodies in the PPa and Hv. These findings suggest that reciprocal connections exist between the NPY neurons and GAL neurons in the brain of the masu salmon. PMID- 19560522 TI - Impulse control disorders in Parkinson's disease in a Chinese population. AB - BACKGROUND: Dopamine agonists have been used as first-line treatments for patients with Parkinson's disease (PD) during its early stage, and several impulse control disorder (ICD) behaviors have been reported to be associated with their use. OBJECTIVE: To investigate the association between ICD behaviors and the use of agonists in Chinese patients with PD and associated risk factors. METHODS: Self-report screening questionnaires were mailed to 400 PD patients treated with anti-parkinsonian drugs in our clinical database and their spouses (served as control group). Those who screened positive for ICD behaviors by questionnaire were further interviewed over the telephone by a movement disorder specialist to confirm the diagnosis. RESULTS: A total of 11 (3.53%) patients were diagnosed with ICD behaviors as follows: lifetime pathological gambling (1, 0.32%); subclinical or clinical hypersexuality (6, 1.92%); binge eating (1, 0.32%); dopamine dysregulation syndrome (2, 0.64%); and compulsive internet browsing (1, 0.32%). ICD behaviors were associated with an increased mean levodopa equivalent daily dosage and alcohol use (p=0.005 and p=0.002, respectively). Patients using dopamine agonists were significantly (p=0.003) more likely to be diagnosed with an ICD (6.3%) as compared to those who were not (0.6%). CONCLUSION: PD patients who took dopamine agonists were more likely to report ICD behaviors in Chinese PD. PMID- 19560523 TI - Phenotypic diversity and expression of GABAergic inhibitory interneurons during postnatal development in lumbar spinal cord of glutamic acid decarboxylase 67 green fluorescent protein mice. AB - The synthesis enzyme glutamic acid decarboxylase (GAD65 or GAD67) identifies neurons as GABAergic. Recent studies have characterized the physiological properties of spinal cord GABAergic interneurons using lines of GAD67-green fluorescent protein (GFP) transgenic mice. A more complete characterization of their phenotype is required to better understand the role of this population of inhibitory neurons in spinal cord function. Here, we characterize the distribution of lumbar spinal cord GAD67-GFP neurons at postnatal days (P) 0, 7, and 14, and adult based on their co-expression with GABA and determine the molecular phenotype of GAD67-GFP neurons at P14 based on the expression of various neuropeptides, calcium binding proteins, and other markers. At all ages >67% of GFP(+) neurons were also GABA(+). With increasing age; (i) GFP(+) and GABA(+) cell numbers declined, (ii) ventral horn GFP(+) and GABA(+) neurons vanished, and (iii) somatic labeling was reduced while terminal labeling increased. At P14, vasoactive intestinal peptide and bombesin were expressed in approximately 63% and approximately 35% of GFP(+) cells, respectively. Somatostatin was found in a small number of neurons, whereas calcitonin gene related peptide never co-localized with GFP. Moderate co-expression was found for all the Ca(2+) binding proteins examined. Notably, most laminae I-II parvalbumin(+) neurons were also GFP(+). Neurogranin, a protein kinase C substrate, was found in approximately 1/2 of GFP(+) cells. Lastly, while only 7% of GFP(+) cells contain nitric oxide synthase (NOS), these cells represent a large fraction of all NOS(+) cells. We conclude that GAD67-GFP neurons represent the majority of spinal GABAergic neurons and that mouse dorsal horn GAD67-GFP(+) neurons comprise a phenotypically diverse population. PMID- 19560525 TI - A new cytotoxic casbane diterpene from Euphorbia pekinensis. AB - A new cytotoxic casbane diterpene, named pekinenal, was isolated from the roots of Euphorbia pekinensis. Its structure was elucidated as 5alpha-hydroxy 1betaH,2alphaH-casba-3Z,7E,11E-triene-18-al by a combination of 1D- and 2D-NMR techniques and confirmed by X-ray crystallography. Pekinenal showed cytotoxic activity against all four human cancer cell lines tested. PMID- 19560526 TI - A shotgun approach to discovering and reconstructing consensus retrotransposons ex novo from dense contigs of short sequences derived from Genbank Genome Survey Sequence database records. AB - Retrotransposons constitute the majority of pseudogenic protein coding regions of most eukaryotic genomes. Most genomes carry tens to thousands of retrotransposon copies derived from dozens of distinct families, but most if not all of these copies are non-functional and contain disabling mutations, including large numbers of indels. Until recently, most regions rich in these elements were virtually ignored in all but the most complete genome sequencing projects, and the full extent of their impact on the structure and function of the genomes of higher eukaryotes was under-appreciated. Even when new retrotransposons are encountered and annotated by automated gene finding programs and similarity searches, coding regions are treated as exons and invariably and not surprisingly mistranslated because of numerous frameshift mutations and large indels. Very few functional retrotransposons contain introns, as in silico annotations imply. While many repetitive DNA consensus sequences have been assembled from collections of largely full-length copies using full-length templates, we have shown that repetitive DNA consensus sequence contigs representing long, moderately high copy-number elements can also be generated ex novo in the absence of templates from very short overlapping sequences. We have devised an in silico strategy to recover and reconstruct consensus sequences of elements up to 20,000 bp by building dense contigs of hundreds of overlapping 400 to 900-bp records found in the Genbank Genome Survey Sequence database. The results are hypothetical ancestral sequences that encode elements that appear to be fully functional with intact open reading frames and other conserved features. PMID- 19560524 TI - Gene delivery to cornea. AB - This paper reviews the strategies of in vivo gene delivery to the cornea. A number of studies have demonstrated the feasibility of targeted delivery of oligonucleotides, small interfering RNA (siRNA), plasmid DNA, and viral vectors to the corneal cells in vivo, specifically stromal keratocytes and corneal epithelial cells, via intrastromal injection, iontophoresis, electroporation, and gene gun. Intrastromal injection of plasmid DNA and adenovirus each can result in efficient transgene expression to stromal keratocytes. The introduction of foreign genes into intact corneal epithelium specifically requires more invasive procedures such as gene gun to disrupt the tight junction barrier and/or cell membranes. The combination of iontophoresis and electroporation was found to be effective in delivering siRNA but not plasmid DNA into the corneal epithelium. Nanocarriers such as polymeric micelles are promising methods of corneal gene delivery. PMID- 19560527 TI - Bisphenol A levels in blood depend on age and exposure. AB - We present two approaches to estimate blood concentrations of Bisphenol A (BPA). Simple kinetic principles were applied to calculate steady state plasma concentrations. A physiologically based model was used to simulate the blood concentration time profile in several age groups exploring the influence of not yet fully developed metabolic capacity on the blood concentrations in the newborn. Both approaches gave concordant results and are in excellent agreement with experimental results [Volkel, W., Colnot, T., Csanady, G.A., Filser, J.G., Dekant, W., 2002. Metabolism and kinetics of bisphenol A in humans at low doses following oral administration. Chem. Res. Toxicol. 15, 1281-1287]. The predictions also agree with published results obtained with a different physiologically based model. According to model simulations, BPA is present in the blood of the normal population at concentrations several orders of magnitude lower than most measurements reported in the literature. At the same external exposure level, the newborn is predicted to have 3 times greater blood concentration than the adult. This is due to the not yet fully developed glucuronidation activity in the newborn, not fully compensated by the unimpaired sulfation pathway. For the highest measured external BPA exposure, the predicted blood concentrations of 2.6 pg/ml (steady state concentration) and 8.2 pg/ml (peak concentration) in the adult are lower than the in vitro concentrations at which inhibiting adiponectin release from human adipocytes and stimulation of beta-cell production and secretion were observed. PMID- 19560528 TI - Vocal singing by prelingually-deafened children with cochlear implants. AB - The coarse pitch information in cochlear implants might hinder the development of singing in prelingually-deafened pediatric users. In the present study, seven prelingually-deafened children with cochlear implants (5.4-12.3 years old) sang one song that was the most familiar to him or her. The control group consisted of 14 normal-hearing children (4.1-8.0 years old). The fundamental frequencies (F0) of each note in the recorded songs were extracted. The following five metrics were computed based on the reference music scores: (1) F0 contour direction of the adjacent notes, (2) F0 compression ratio of the entire song, (3) mean deviation of the normalized F0 across the notes, (4) mean deviation of the pitch intervals, and (5) standard deviation of the note duration differences. Children with cochlear implants showed significantly poorer performance in the pitch-based assessments than the normal-hearing children. No significant differences were seen between the two groups in the rhythm-based measure. Prelingually-deafened children with cochlear implants have significant deficits in singing due to their inability to manipulate pitch in the correct directions and to produce accurate pitch height. Future studies with a large sample size are warranted in order to account for the large variability in singing performance. PMID- 19560529 TI - Effect of lavender oil on motor function and dopamine receptor expression in the olfactory bulb of mice. AB - AIM OF THE STUDY: Although treatment with the essential oil of lavender induces neuroemotional changes, there is a lack of data regarding its specific effects on neurotransduction, especially dopaminergic neurotransduction. We investigated the relationship between altered motor activity and changes in the expression of dopamine receptors (DR), particularly the receptor subtypes D2 and D3, in lavender oil-treated mice. MATERIALS AND METHODS: After the administration of lavender oil (intraperitoneal injections of 10-1000 mg/kg lavender oil once per day for 5 days), motor coordination and dopamine receptor expression were examined in the olfactory bulb and the striatum of the mouse brain. RESULTS: After 5 days, mice treated with 1000 mg/kg lavender oil showed significantly increased rotarod activity when compared to controls. Although DRD2 expression showed no change in the olfactory bulb or striatum of lavender-treated mice, DRD3 expression increased significantly in the olfactory bulb; this increase was dose dependent and was observed at both the mRNA and protein levels. CONCLUSIONS: These data indicate that altered dopamine D3 receptor subtype homeostasis in the olfactory bulb may contribute to lavender oil-induced behavioral change. PMID- 19560530 TI - Triptolide promotes generation of FoxP3+ T regulatory cells in rats. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Triptolide (TPT), a component of the Chinese herb Triptergium wilfordii, has potent immunosuppressive and anti-inflammatory activity and is used clinically in recipients of kidney transplantation. AIM OF THE STUDY: This work aimed to investigate the effect of TPT on the differentiation of regulatory T lymphocytes (Tregs) from CD4+ cells in rats. MATERIALS AND METHODS: MACS-purified rat CD4+ cells were costimulated with anti CD3 and anti-CD28 in the presence of TGF-beta to induce the expression of FoxP3, which was detected by flow cytometry. TPT and cyclosporine A (CsA) were separately added into the cultures to observe the effect on the expression of FoxP3. Kidney transplantation was performed in rats that either received no treatment or were treated with TPT after transplantation. RESULTS: TPT treatment enhanced the expression of FoxP3 in CD4+ cells, whereas CsA inhibited the FoxP3 expression. In the rat kidney transplantation model, the recipient rats treated with TPT survived longer than the control rats (18-19.83 vs 6.83 days, P<0.05). Meanwhile, the FoxP3+ T cells in the spleens of treated rats were higher than those from the untreated rats (12.4% vs 4.7%, P<0.05). CONCLUSIONS: These data suggest that TPT may promote the differentiation of CD4+ cells to FoxP3+ Tregs. This would be at least one of the pathways responsible for the immunosuppressive activity of TPT. PMID- 19560531 TI - Experimental evidence of the anaphrodisiac activity of Humulus lupulus L. in naive male rats. AB - ETHNOPHARMACOLOGICAL RELEVANCE: In the folk medicine Humulus lupulus L. (hops) is mainly recommended as a mild sedative with antispasmodic and digestive properties. It is also reputed to exert an anaphrodisiac effect but it is still lacking the experimental evidence of this activity. AIM OF THE STUDY: To evaluate the influence of Humulus lupulus extract on sexual behavior of both naive and sexually potent male rats; thereafter to investigate the role of 8 prenylnarigenin (8-PN) in the effect displayed by the hop extract. MATERIALS AND METHODS: Sprague-Dawley male rats both naive and sexually potent were acutely administered with the hop extract dosed at 5, 10, 25 and 50 mg/kg. In addition the extract was administered daily for 10 consecutive days at the dose of 0.25 mg/kg/day in sexually potent animals. The pure compound 8-PN was acutely administered in naive rats at the dosages of 5, 12.5 and 25 microg/kg. All the animals were screened for their sexual behavior manifestation during the mating test. RESULTS: In naive rats the acute administration of Humulus lupulus extract at the doses of 25 and 50 mg/kg significantly reduced the percentage of mounting and ejaculating animals, in comparison to vehicle controls. The other parameters recorded during the mating test were not affected by the hop extract. In sexually potent rats nor the acute neither the repeated administration of the extract modified their copulatory behavior. The pure compound 8-PN failed to influence male sexual behavior of naive rats. CONCLUSION: Humulus lupulus extract exerted an anaphrodisiac effect only in naive rats by inhibiting their mounting and ejaculating behavior. The presence of 8-PN in the extract could be only partially involved in the observed anaphrodisiac effect. PMID- 19560532 TI - Antioxidant and lipoxygenase inhibitory activity of oligostilbenes from the leaf and stem of Vitis amurensis. AB - ETHNOPHARMACOLOGICAL RELEVANCE: The root and stem of Vitis amurensis (Vitaceae) have popularly used as traditional medicine for treatment of cancer and various pains in Korea and Japan. Recent studies, its root and stem possess anti inflammatory, anti-tumor activities, and protective effects against beta-amyloid induced oxidative stress. AIM OF THE STUDY: This study deals with the isolation, structural identification of the potent bioactive compounds from the leaf and stem, and their antioxidant capacity, as well as anti-inflammatory effect via lipoxygenase inhibitory assay. MATERIALS AND METHODS: All isolated compounds yielded after using column chromatography were identified base on the physico chemical properties and 1D, 2D NMR spectra. The scavenge ability against DPPH and ABTS(+) radicals, and to inhibit lipid peroxidation, as well as lipoxygenase type I inhibitory activity of all isolates were performed using in vitro assays. RESULTS: Eleven resveratrol derivatives (1-11), including a new oligostilbene cis amurensin B (9), whose structures were determined on the basis of extensively spectral analyses, were isolated from the leaf and stem of Vitis amurensis. The isolates (1-11) were examined for their antioxidant activities by evaluating scavenge ability against DPPH and ABTS(+) radicals, and to inhibit lipid peroxidation. Stilbenes 1 and 4, and oligostilbenes 5-10 displayed moderate anti lipid peroxidation activities, but all the isolates exhibited strong ABTS(+) radical scavenging activity in the dose-dependent manner. In addition, the isolates showed stronger inhibitory capacity against soybean lipoxygenase type I than that of baicalein, a positive control. Of the isolates, r-2-viniferin (8) exhibited the strongest scavenging activity against ABTS(+) radical with TEAC value of 5.57, and the most potential inhibitory effect on soybean lipoxygenase with the IC(50) value of 6.39 microM. CONCLUSION: This is the first report on the potential antioxidant and LOX-1 inhibitory effects of oligostilbenes isolated from the leaf and stem of Vitis amurensis. In addition, chemical compositions isolated from the leaf and stem are almost similar to those isolated from the root of Vitis amurensis. Therefore, the results may explain, in part, the uses of the leaf and stem, as well as the root of Vitis amurensis in the Korean traditional medicine. PMID- 19560533 TI - Altered levels of synapsin I, dopamine transporter, dynorphin A, and neuropeptide Y in the nucleus accumbens and striatum at post-puberty in rats treated neonatally with pregnenolone or DHEA. AB - It is well documented that neonatal neurosteroid administration influences brain development. In our previous studies, administration of pregnenolone, the precursor of neurosteroids, during the neonatal period altered the activity of dopamine (DA) in the striatum. Furthermore, neonatal treatment with pregnenolone or dehydroepiandrosterone (DHEA) increased synapse-related protein synapsin I as well as neuropeptide Y (NPY) in the hippocampus. The present study examined the effects of neonatal treatment with pregnenolone or DHEA on synapsin I, DA transporter (DAT), dynorphin A, and NPY in the striatum and the core and shell of the nucleus accumbens at post-puberty. Administration of pregnenolone or DHEA during the neonatal period increased immunodensity of synapsin I in the dorsomedial or ventrolateral striatum. DAT immunodensity in the striatum and the nucleus accumbens core as well as dynorphin A immunodensity in the nucleus accumbens core were increased in DHEA-treated but not in pregnenolone-treated rats. In addition, the size, but not numbers, of NPY-positive cells in the nucleus accumbens core was increased in pregnenolone- and DHEA-treated rats. The results suggest that neurosteroid levels during the neonatal period have larger impact on synaptic formation, development of DA and NPY systems in the nigrostriatal rather than the mesolimbic pathway. PMID- 19560534 TI - Gingko biloba leaf extract induces oxidative stress in carcinoma HSC-2 cells. AB - The antiproliferative effects of a Gingko biloba leaf extract to cells from tissues of the human oral cavity were studied. Toxicity to carcinoma HSC-2 cells was correlated with the prooxidative nature of the extract. G. biloba leaf extract generated reactive oxygen species (ROS) in cell culture medium and, albeit to a lesser extent, in buffer, with higher levels detected at alkaline pH. Lowered levels of ROS were detected in culture medium coamended with the extract and with either catalase or superoxide dismutase, indicating the generation of hydrogen peroxide and superoxide anion, respectively. Biological activity of the extract was through oxidative stress. Toxicity to the HSC-2 cells was lessened by the ROS scavengers, divalent cobalt and pyruvate, by catalase, and by the antioxidant, N-acetyl-L-cysteine, and was potentiated by the glutathione depleters, DL-buthionine-[S,R]-sulfoximine, 1-chloro-2,4-dinitrobenzene, and bis(2-chloroethyl)-N-nitrosourea. G. biloba reacted directly with authentic glutathione and lowered the intracellular glutathione content in HSC-2 cells. Induction of apoptosis upon exposure of HSC-2 cells to G. biloba extract was noted by apoptotic cell morphologies, by TUNEL staining, and by PARP cleavage. The data strongly suggest that the prooxidative nature of the G. biloba extract was the cause of apoptotic cell death. PMID- 19560535 TI - Fatigue in primary Sjogren's syndrome--a link to sickness behaviour in animals? AB - Interleukin-1beta (IL-1beta) is involved in the regulation of sickness behaviour in response to infection and inflammation in animals. Human fatigue can be considered an element of sickness behaviour and is a prominent and often disabling phenomenon in autoimmune diseases such as primary Sjogren's syndrome (PSS). The role of the IL-1 system in the fatigue of patients with PSS was explored. A cerebrospinal fluid (CSF) analysis of IL-1beta, IL-1Ra, and IL-1sRII was performed in 54 PSS patients and 53 control subjects. Fatigue was evaluated in the patients using the Fatigue Severity Scale (FSS) and a fatigue visual analogue scale (VAS); mood was evaluated using the Beck Depression Inventory (BDI). There were higher CSF levels of IL-1Ra pg/mL in PSS patients vs. controls (median 38.4: range 15.4-81.7 vs. 33.7: 7.3-163.1, p=0.026). Fatigue VAS scores were associated with increasing CSF levels of IL-1Ra in PSS patients (R(2)=0.11, p=0.015). In a subgroup analysis of the non-depressed PSS patients (N=37; 69%), the association between VAS scores and IL-1Ra was even stronger (R(2)=0.20, p=0.006). The positive association between VAS scores and IL-1Ra remained significant in a multiple regression analysis adjusting for age and BDI scores. Increased levels of IL-1Ra in the CSF are associated with increasing fatigue in PSS patients, indicating that the activated IL-1 system is a possible biological factor associated with fatigue. PMID- 19560537 TI - Improvement of cyclosporine A bioavailability by incorporating ethyl docosahexaenoate in the microemulsion as an oil excipient. AB - The aims of this study were to determine the effect of ethyl docosahexaenoate (DHA-EE) on cyclosporine A (CsA) bioavailability, while also examining the effect of DHA-EE on CsA when DHA-EE was incorporated into a microemulsion formulation as an oil ingredient. The oral co-administration of DHA-EE and CsA increased the blood CsA concentration in a dose-dependent manner, and the AUC and C(max) both increased by about 2-fold with 100mg/kg DHA-EE. The microemulsion formulation of CsA consisted of Tween-20, ethanol, water, and DHA-EE (53.3/6.5/35.9/3.3 w/w%) (namely DHA-ME) was transparent and stable with an average particle size of 50 nm, which was similar to that of the control formulation incorporating vitamin E instead of DHA-EE (namely VE-ME). The permeabilities of CsA from DHA-ME, VE-ME and Neoral formulations across an artificial membrane were not significantly different. The C(max) and AUC(0-infinity) of CsA in rats administered DHA-ME significantly increased by approximately 2-fold in comparison to that of VE-ME. The relative oral bioavailability (F(r)) of DHA-ME in comparison to Neoral was determined to be 114%, while the F(r) of VE-ME was only 60%. It was, therefore, suggested that the use of DHA-EE as an oil excipient may be promising for the development of a microemulsion formulation of CsA with an improved oral bioavailability. PMID- 19560538 TI - Lipid nanocapsules: ready-to-use nanovectors for the aerosol delivery of paclitaxel. AB - Aerosol drug delivery permits the development of dose-intensification strategies in severe, malignant lung diseases. The aim of the study was to demonstrate that the encapsulation of paclitaxel in lipid nanocapsules (LNCs), a novel drug nanocarrier for lipophilic components, allows one to provide pulmonary drug delivery of paclitaxel by nebulisation, thereby allowing preclinical and clinical studies. LNC dispersions are made into aerosols with commercial nebulisers. The structure, drug payload and cytotoxicity of nebulised LNCs were compared to fresh LNCs. The results demonstrated that LNC dispersions could be made into aerosols by using mesh nebulisers without altering the LNC structure. Only eFlow rapid produced aerosols are compatible with human use: the mean duration to nebulise 3 ml of LNC dispersion is less than 9 min, with an aerosol mass median aerodynamic diameter equal to 2.7+/-0.1 microm and a fine-particle fraction (between 1.0 and 5.0 microm) of 81.5+/-3.1%. No modifications of drug payload or cytotoxicity effects of paclitaxel-loaded LNC (PTX-LNC) were observed. In order to carry out preclinical studies, a scaled-up LNC formulation protocol was used. Chemical parameters, such as acidity and osmolarity, were optimised, and a storage procedure for PTX-LNC batches was set-up. Animal studies are now needed to determine the tolerance and therapeutic potential of LNC dispersion aerosols. PMID- 19560536 TI - A conserved hydrophobic surface of the LARG pleckstrin homology domain is critical for RhoA activation in cells. AB - Leukemia associated Rho guanine nucleotide exchange factor (LARG) activates RhoA in response to signals received by specific classes of cell surface receptors. The catalytic core of LARG is a Dbl homology (DH) domain whose activity is modulated by an adjacent pleckstrin homology (PH) domain. In this study, we used a transcriptional assay and confocal microscopy to examine the roles of several novel structural features of the LARG DH/PH domains, including a conserved and exposed hydrophobic patch on the PH domain that mediates protein-protein interactions in crystal structures of LARG and its close homolog PDZ-RhoGEF. Mutation of the hydrophobic patch has no effect on nucleotide exchange activity in vitro, but abolished the ability of LARG to activate RhoA and to induce stress fiber formation in cultured cells. The activity of these mutants could be rescued by fusion with exogenous membrane-targeting domains. However, because membrane recruitment by activated G alpha(13) subunits was not sufficient to rescue activity of a hydrophobic patch mutant, the LARG PH domain cannot solely contribute to membrane targeting. Instead, it seems likely that the domain is involved in regulatory interactions with other proteins near the membrane surface. We also show that the hydrophobic patch of the PH domain is likely important for the activity of all Lbc subfamily RhoGEFs. PMID- 19560539 TI - Xylan decomposition by Aspergillus clavatus endo-xylanase. AB - Agricultural and forest waste products are abundant and low-cost biomass sources useful in renewable fuel energy and feedstock preparation. Hydrolysis of a major biomass component, hemicellulose, is accomplished by the action of endo xylanases. Reaction products vary in composition and degree of polymerization as a function of both feedstock and the enzyme activities utilized, ranging from monomeric sugars to complex branched polysaccharides. The study herein describes heterologous expression in Aspergillus awamori of a betabeta-(1-4) endo-xylanase isolated from the whole-genome DNA sequence of A. clavatus along with a comprehensive biochemical and functional analysis of the enzyme, including substrate preference and hydrolysis patterns. The A. clavatus xylanase promotes incomplete hydrolysis of xylan substrates resulting in xylobiose, xylotriose and xylotetraose. Incomplete degradation resulting in xylo-oligomers is appealing for functional foods as the beneficial effect of oligosaccharides on gastrointestinal micro flora includes preventing proliferation of pathogenic intestinal bacteria and facilitating digestion and absorption of nutrients. PMID- 19560540 TI - A nucleoprotein-hairpin in transcription regulation. AB - Regulation of gene transcription in both prokaryotes and eukaryotes involves the formation of DNA-multiprotein complexes. These complexes build a precise three dimensional topology allowing communication between distal regions of DNA. The switch from early to late transcription in bacteriophage O29 involves binding of viral proteins, p4 and p6, to a region of the genome containing the early promoters A2c and A2b and the late promoter A3. Atomic force microscopy imaging under aqueous buffering conditions of complexes built after DNA incubation with proteins p4 and p6 shows the formation of a nucleoprotein arrangement with consistent morphology. These two low specificity DNA binding proteins are capable of bending 160 base pairs into a nucleoprotein-hairpin stable enough to be imaged by AFM. The functional implications of this nucleoprotein-hairpin in the coordinated regulation of early and late promoters are discussed. PMID- 19560541 TI - DNA-tethered membranes formed by giant vesicle rupture. AB - We have developed a strategy for preparing tethered lipid bilayer membrane patches on solid surfaces by DNA hybridization. In this way, the tethered membrane patch is held at a controllable distance from the surface by varying the length of the DNA used. Two basic strategies are described. In the first, single stranded DNA strands are immobilized by click chemistry to a silica surface, whose remaining surface is passivated to prevent direct assembly of a solid supported bilayer. Then giant unilamellar vesicles (GUVs) displaying the antisense strand, using a DNA-lipid conjugate developed in earlier work [Chan, Y. H.M., van Lengerich, B., et al., 2008. Lipid-anchored DNA mediates vesicle fusion as observed by lipid and content mixing. Biointerphases 3 (2), FA17-FA21], are allowed to tether, spread and rupture to form tethered bilayer patches. In the second, a supported lipid bilayer displaying DNA using the DNA-lipid conjugate is first assembled on the surface. Then GUVs displaying the antisense strand are allowed to tether, spread and rupture to form tethered bilayer patches. The essential difference between these methods is that the tethering hybrid DNA is immobile in the first, while it is mobile in the second. Both strategies are successful; however, with mobile DNA hybrids as tethers, the patches are unstable, while in the first strategy stable patches can be formed. In the case of mobile tethers, if different length DNA hybrids are present, lateral segregation by length occurs and can be visualized by fluorescence interference contrast microscopy making this an interesting model for interactions that occur in cell junctions. In both cases, lipid mobility is high and there is a negligible immobile fraction. Thus, these architectures offer a flexible platform for the assembly of lipid bilayers at a well-defined distance from a solid support. PMID- 19560542 TI - Characterization, expression, and response to stress of p8 gene in amphioxus. AB - The amphioxus Branchiostoma belcheri tsingtaunese homolog of p8, Bbp8, was identified from the gut cDNA library. The full-length Bbp8 cDNA consists of 1032 bp, which is clearly longer than those of p8 in human, mouse, rat, frog, zebrafish and fruit fly. The genomic DNA sequences of amphioxus p8 contain three exons and two introns, which is similar to the exon/intron organization of p8 homologues in vertebrates such as human, mouse and zebrafish, while it is sharply different to the organization of p8 gene in fruit fly, which has only one exon. Sequence alignment and phylogenetic analysis showed that the basic helix-loop helix (bHLH) region of p8 is well conserved during the long process of evolution, and Bbp8 is more close to its homologous proteins in the invertebrates than to those in the vertebrates. RT-PCR and In situ hybridization histochemistry demonstrated the expression of Bbp8 in all the tissues assayed, with relatively higher expression in the gut, gill and ovaries. Quantitative real-time PCR assay revealed quick up-regulation of Bbp8 transcripts on lipopolysaccharide (LPS) challenge and starvation, implying a stress-related function for Bbp8. PMID- 19560543 TI - Different roles of cytoarchitectonic BA 44 and BA 45 in phonological and semantic verbal fluency as revealed by dynamic causal modelling. AB - The interactions of left cytoarchitectonic BA 44 and BA 45 during semantic and phonological verbal fluency tasks were investigated using dynamic causal modelling (DCM). Three different models were tested, all of which featured BA 44 and BA 45 as top-down driven interconnected nodes projecting to the motor cortex as the final output region. Model #1 represents the hypothesis that BA 45 is involved in lexical retrieval including both semantic and phonological processes, while BA 44 supports other phonological processes. Model #2 reflects the notion of a clear-cut segregation of computational processes sustained by BA 44 (phonological processing) and BA 45 (semantic processing). Model #3 was based on the hypothesis that both BA 44 and BA 45 support semantic and phonological processing. When these models were compared against each other by Bayesian model selection, evidence emerged in favour of the first model, implying that BA 45 supports word retrieval processes whereas BA 44 is involved in processing phonological information during word generation. In a subsequent analysis of the derived model parameters for model #1, all connection strengths were significantly positive except for the inhibitory coupling between BA 44 and BA 45. This inhibition may reflect how the phonological analysis in BA 44 during word generation constrains lexical word retrieval in BA 45. To conclude, DCM provided additional insights into the roles of BA 44 and BA 45 during verbal fluency revealing the involvement of BA 45 in lexical retrieval and the relevance of BA 44 for phonological processing during word generation. PMID- 19560544 TI - Neural substrates of semantic relationships: common and distinct left-frontal activities for generation of synonyms vs. antonyms. AB - Synonymous and antonymous relationships among words may reflect the organization and/or processing in the mental lexicon and its implementation in the brain. In this study, functional magnetic resonance imaging (fMRI) is employed to compare brain activities during generation of synonyms (SYN) and antonyms (ANT) prompted by the same words. Both SYN and ANT, when compared with reading nonwords (NW), activated a region in the left middle frontal gyrus (BA 46). Neighboring this region, there was a dissociation observed in that the ANT activation extended more anteriorly and laterally to the SYN activation. The activations in the left middle frontal gyrus may be related to mental processes that are shared in the SYN and ANT generations, such as engaging semantically related parts of mental lexicon for the word search, whereas the distinct activations unique for either SYN or ANT generation may reflect the additional component of antonym retrieval, namely, reversing the polarity of semantic relationship in one crucial dimension. These findings suggest that specific components in the semantic processing, such as the polarity reversal for antonym generation and the similarity assessment for synonyms, are separately and systematically laid out in the left-frontal cortex. PMID- 19560545 TI - Different representations of relative and absolute subjective value in the human brain. AB - Relative reward value is important for the choice between a set of available rewards, and absolute reward value for stable and consistent economic choice. It is unclear whether in the human brain subjective absolute value representations can be dissociated from relative reward value representations. Using fMRI, we investigated how the subjective pleasantness of an odor is influenced by whether the odor is presented in the context of a relatively more pleasant or less pleasant odor. We delivered two of a set of four odors separated by a delay of 6 s, with the instruction to rate the pleasantness of the second odor, and searched for brain regions where the activations were correlated with the absolute pleasantness rating of the second odor, and for brain regions where the activations were correlated with the difference in pleasantness of the second from the first odor, that is, with relative pleasantness. Activations in the anterolateral orbitofrontal cortex tracked the relative subjective pleasantness, whereas activations in the anterior insula tracked the relative subjective unpleasantness. In contrast, in the medial and midorbitofrontal cortex activations tracked the absolute pleasantness of the stimuli. Thus, both relative and absolute subjective value signals which provide important inputs to decision making processes about which stimulus to choose are separately and simultaneously represented in the human brain. PMID- 19560546 TI - The influence of context valence in the neural coding of monetary outcomes. AB - The emotional significance of objects and events depends on the context in which they occur. Using functional magnetic resonance imaging, we examined the modulation of neural responses to monetary outcomes while subjects performed a decision-making task in a positive and a negative economic context. Neural responses indicated a relative regional specialization in the neural coding of outcome valence and followed three distinct patterns. The nucleus accumbens (NAc) and orbital frontal cortex (OFC) appeared to code the most extreme outcome in each context, with a potentiated response for favorable outcomes by a positive context. The amygdala and insula appeared to also code highly salient outcomes, but showed a potentiated response to unfavorable outcomes occurring in a negative context. The medial prefrontal cortex (medPFC), on the other hand, only coded favorable responses occurring in a positive context. Moreover, the medPFC showed large inter-individual variability when responding to outcomes in a negative context, suggesting that its role in a negative context may depend on a number of individual factors. The results of this work provide evidence of complex valence based regional dissociations that are influenced by contextual factors. PMID- 19560547 TI - Alterations in neural connectivity in preterm children at school age. AB - Converging data suggest recovery from injury in the preterm brain. We used functional magnetic resonance imaging (fMRI) to test the hypothesis that cerebral connectivity involving Wernicke's area and other important cortical language regions would differ between preterm (PT) and term (T) control school age children during performance of an auditory language task. Fifty-four PT children (600-1250 g birth weight) and 24 T controls were evaluated using an fMRI passive language task and neurodevelopmental assessments including: the Wechsler Intelligence Scale for Children - III (WISC-III), the Peabody Individual Achievement Test - Revised (PIAT-R) and the Peabody Picture Vocabulary Test - Revised (PPVT-R) at 8 years of age. Neural activity was assessed for language processing and the data were evaluated for connectivity and correlations to cognitive outcomes. We found that PT subjects scored significantly lower on all components of the WISC-III (p<0.009), the PIAT-R Reading Comprehension test (p=0.013), and the PPVT-R (p=0.001) compared to term subjects. Connectivity analyses revealed significantly stronger neural circuits in PT children between Wernicke's area and the right inferior frontal gyrus (R IFG, Broca's area homologue) and both the left and the right supramarginal gyri (SMG) components of the inferior parietal lobules (p15, 22% of patients were reclassified to a higher risk with an event rate of 44% compared with 10% when the score was 13GPa and a maximum fracture toughness of 6.9MPam(-1/2), with an average grain size of 0.4+/ 0.17microm. The observed crack deflection was an important mechanism in the improved fracture toughness of the composite. In addition, the grain size and residual porosity also seem to be factors in obtaining a wide range of hardness as well as fracture toughness by varying the Al(2)O(3) whiskers and ZrO(2) (TZ 3Y) content. The use of alumina-whisker-reinforced composites in dental applications could be promising for increasing hardness and fracture toughness compared with other materials. The reported values for these composites can compete with those of commercially available materials in different dental applications. PMID- 19560563 TI - Embryonic emergence of the respiratory rhythm generator. AB - Breathing is vital for life ex utero and therefore requires that the respiratory rhythm generator (RRG), the central neural network generating the continuous rhythmic motor command, be functional at birth. The RRG, located in the brainstem, appears to comprise two interacting respiratory oscillators: the parafacial respiratory group (pFRG), and the preBotzinger complex (preBotC). Data on the establishment of these respiratory oscillators during embryonic and foetal periods are beginning to be produced. The present paper provides a short review of the current knowledge regarding: (i) the emergence of activity in the two respiratory oscillators and (ii) their functional coupling during prenatal development in rodents. PMID- 19560565 TI - Surgical approach to hydrocarbon injection in upper extremities: case series. AB - BACKGROUND: Hydrocarbon injection, which mostly occurs in the upper extremity, is commonly seen in domestic and industrial accidents, or attempted suicide. Surgical approach to chemical cellulites and other complications in these patients is controversial. METHOD: This survey has been carried out on 21 patients admitted to general surgery unit of Loqman Hakim hospital, Tehran, Iran with the complaint of hydrocarbon injection in 2001-2005. RESULTS: The most common injected material was petroleum (in 8 patients). Anterior forearm was the most common injection site (in 16 patients). All of the patients had inflammation, swelling, and localized tenderness. Leukocytosis was present in 18 of 21 patients. Compartment syndrome happened in 13 patients; 5 with necrotizing fasciitis. All of the patients with compartment syndrome underwent fasciotomy and debridement. Split thickness skin graft was done for 10 patients after 2 weeks of admission. There was no mortality or limb loss during the study period. CONCLUSIONS: Prevention of the systemic complications, hospitalization, close monitoring and treatment of chemical cellulites are the main parts of the management of patients with hydrocarbon injection. Surgical intervention is recommended when compartment syndrome or other local complications occur. PMID- 19560566 TI - Should radiocontrast be diluted for operative cholangiography? PMID- 19560567 TI - Statistical illiteracy is damaging our health: doctors and patients need to understand numbers if meaningful dialogues are to occur. PMID- 19560568 TI - Characterization of the region of the aryl hydrocarbon receptor required for ligand dependency of transactivation using chimeric receptor between Drosophila and Mus musculus. AB - The aryl hydrocarbon receptor (AhR) is a ligand-activated transcriptional factor. Although 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is high affinity and toxic to many vertebrate animals, invertebrate AhRs including Drosophila melanogaster AhR (spineless) have no ability to bind exogenous chemicals as ligands. To analyze the ligand-binding domain (LBD) of AhR, we used chimeras between mouse and Drosophila AhR. The chimeric AhR revealed that the LBD determines constitutive transactivation in Drosophila AhR or ligand-dependent activation in mouse AhR. The LBD was further divided into three blocks that corresponded to amino acids 230-300, 301-361, and 361-420 of the mouse sequence. Six chimeric proteins clarified that amino acids 291-350 of the Drosophila LBD, i.e. the middle region, were required to keep the protein in the active form in the absence of ligand binding, whereas in the mouse AhR, this region was required to maintain the protein in the inactive form in the absence of ligand. Furthermore, Arg346 in the middle region of the mouse LBD, was identified as amino acids that were critical for AhR activation by site-directed mutagenesis. PMID- 19560569 TI - T-cell leukemia translocation-associated gene (TCTA) protein is required for human osteoclastogenesis. AB - Synovial tissues of patients with rheumatoid arthritis (RA) include factors regulating bone resorption, such as receptor activator NF-kappaB ligand (RANKL), TNFalpha, IL-6, IL-17 and IFNgamma. However, in addition to these cytokines, other factors expressed in synovial tissues may play a role in resorbing bone. Here, our objective was to identify novel proteins expressed in synovial tissues of RA that regulate human osteoclastogenesis. Proteins were purified from synovial tissues of patients with RA, using gel filtration chromatography, ion exchange chromatography, reverse-aspect HPLC, and mass spectrometry. We evaluated the effects of the purified fractions on human osteoclastogenesis induced by RANKL and M-CSF. We determined the amino acid sequences showing inhibitory activity on human osteoclastogenesis. In addition, we synthesized novel peptides from the molecule including the amino acid sequences. Then, we evaluated the effects of the peptides and antibodies against the molecule on human osteoclastogenesis from monocytes and mature osteoclasts, and on pit formation by mature osteoclasts using Osteologic discs. We examined the effect of the peptide on the expression of both mRNA and protein of NFATc1. We also examined the effect of RANKL on the expression of mRNA of the molecule on osteoclasts and macrophages. We identified a small peptide including Gly-Gln-Asn (GQN) with inhibitory activity on human osteoclastogenesis. We then found that GQN is included in the amino acid sequence of the extra-cellular domain of TCTA protein, which is expressed ubiquitously in normal human tissues, but whose function has not been clarified. We designed novel peptides, including GQN, from the sequence of TCTA protein. One of these peptides (29-mer), but not a scrambled peptide for the 29-mer peptide, potently inhibited RANKL-induced human osteoclastogenesis. The peptide also inhibited pit formation of mature human osteoclasts and suppressed the formation of large osteoclasts in the culture of mature osteoclasts. Furthermore, polyclonal antibodies against TCTA protein suppressed the formation of large osteoclasts in the cultures of both monocytes and mature osteoclasts, supporting our hypothesis. Peptide A did not significantly inhibit the expression of both mRNA and protein of NFATc1 in osteoclasts. Our novel peptide and polyclonal antibodies against the peptide inhibited human osteoclastogenesis and the function of mature osteoclasts, preventing cellular fusion by TCTA protein and a putative counterpart molecule. PMID- 19560571 TI - Reply of the authors to the letter by Broek et al. on June 9, 2008. PMID- 19560572 TI - The role of the endogenous opioid system in polycystic ovary syndrome. AB - OBJECTIVE: To review the complex role of the opioid system in reproduction and carbohydrate metabolism, abnormalities in the opioid system in women with polycystic ovary syndrome (PCOS), and the role of opioid antagonists in the management of PCOS-related infertility. DESIGN: Pertinent articles were identified through a computer PubMed search. References of selected articles were hand searched for additional citations. CONCLUSION(S): Endogenous opioids are generally considered inhibitory central neurotransmitters. Peripherally, opioids are involved in the regulation of pancreatic islet function, hepatic insulin clearance, and glucose metabolism, potentially contributing to the pathogenesis of hyperinsulinemia and insulin resistance in PCOS. The presence of sex steroids is required for normal function of the opioid system in both GnRH secretion and carbohydrate metabolism. In women with PCOS, growing evidence suggests dysregulation of the opioid system both centrally and peripherally, with complex interactions. The opioid system effects on carbohydrate metabolism appear to be modulated by obesity. Finally, naltrexone has been demonstrated to successfully augment traditional ovulation induction regimens, but has limited support as a single ovulation induction agent for PCOS. PMID- 19560570 TI - Beta-arrestin2 regulates parathyroid hormone effects on a p38 MAPK and NFkappaB gene expression network in osteoblasts. AB - Interaction of the cytoplasmic adaptor molecule beta-arrestin2 with the activated parathyroid hormone (PTH)/PTHrP receptor inhibits G protein mediated signaling and triggers MAPKs signaling. In turn, the effects of both intermittent (i.) and continuous (c.) PTH on bone are altered in beta-arrestin2-deficient (Arrb2(-/-)) mice. To elucidate the expression profile of bone genes responsive to PTH and targeted for regulation by beta-arrestin2, we performed microarray analysis using total RNA from primary osteoblastic cells isolated from wild-type (WT) and Arrb2( /-) mice. By comparing gene expression profiles in cells exposed to i.PTH, c.PTH or vehicle (Veh) for 2 weeks, we found that i.PTH specifically up-regulated 215 sequences (including beta-arrestin2) and down-regulated 200 sequences in WT cells, about two-thirds of them being under the control of beta-arrestin2. In addition, beta-arrestin2 appeared necessary to the down-regulation of a genomic cluster coding for small leucin-rich proteins (SLRPs) including osteoglycin, osteomodulin and asporin. Pathway analyses identified a main gene network centered on p38 MAPK and NFkappaB that requires beta-arrestin2 for up- or down regulation by i.PTH, and a smaller network of PTH-regulated genes centered on TGFB1, that is normally repressed by beta-arrestin2. In contrast the expression of some known PTH gene targets regulated by the cAMP/PKA pathway was not affected by the presence or absence of beta-arrestin2 in osteoblasts. These results indicate that beta-arrestin2 targets prominently p38 MAPK- and NFkappaB-dependent expression in osteoblasts exposed to i.PTH, and delineates new molecular mechanisms to explain the anabolic and catabolic effects of PTH on bone. PMID- 19560573 TI - Cervical intramural ectopic pregnancy. AB - OBJECTIVE: To describe a cervical intramural ectopic pregnancy. DESIGN: Case report. SETTING: Maternity hospital. PATIENT(S): A 28-year-old multiparous woman was seen with vaginal bleeding and a positive urinary pregnancy test after delayed menstruation. Ultrasonography and pelvic examination revealed an empty uterine cavity and a hyperechoic focus within an enlarged cyanotic anterior cervical lip. Vaginal bleeding was controlled with aspiration curettage; however, macroscopic and ultrasonographic views of the cervix remained the same. One week later the anterior cervical lip was found to be ruptured. INTERVENTION(S): Decidual tissue was removed from the ruptured area. MAIN OUTCOME MEASURE(S): Histopathologic confirmation of chorionic villi and presence of a connection between the endocervix and the ruptured area. RESULT(S): No connection was found with the endocervical channel when controlled with a 1-mm Hegar uterine dilator, and histopathologic examination of removed materials revealed chorionic villi. CONCLUSION(S): The cervical intramural ectopic pregnancy is an extraordinary clinical situation and might lead to diagnostic and therapeutic challenges. PMID- 19560574 TI - American Society for Reproductive Medicine on the move: 2008 President's Address. PMID- 19560576 TI - Breakdown in epithelial barrier function in patients with asthma: identification of novel therapeutic approaches. AB - The bronchial epithelium is pivotally involved in the provision of chemical, physical, and immunologic barriers to the inhaled environment. These barriers serve to maintain normal homeostasis, but when compromised, the immunologic barrier becomes activated to protect the internal milieu of the lung. We discuss what is currently understood about abnormalities in these barrier functions in patients with asthma and consider novel therapeutic opportunities that target this key structure. PMID- 19560579 TI - Sublingual immunotherapy, meta-analysis, and knowledge in the age of information. PMID- 19560577 TI - Alterations in epithelial barrier function and host defense responses in chronic rhinosinusitis. AB - Chronic rhinosinusitis (CRS) is characterized by a chronic symptomatic inflammation of the nasal and paranasal sinus mucosae and is one of the most frequently reported chronic diseases in the United States, with an estimated prevalence of greater than 10% of the general population. Although the pathogenesis of CRS remains poorly understood, there is evidence for a role of bacteria and fungi, as well as the presence of a robust adaptive immune response in the upper airways and sinuses. Recent studies of CRS, as well as several other diseases in the skin and respiratory epithelium, have uncovered evidence that deficiencies in epithelial immune barrier function might compromise the interaction between the host and external immune stimuli. Recent studies suggest the hypothesis that reduced expression of antimicrobial S100 proteins, particularly psoriasin and calprotectin, might lead to increased susceptibility to bacterial and fungal colonization in patients with CRS. The main emphasis of this review will be to highlight the current literature that suggests that a defect in the expression of a broad set of epithelially derived genes might lead to barrier compromise and subsequently a dysfunctional host immune response to environmental agents in patients with CRS. PMID- 19560580 TI - Alcohol and harm reduction in Russia. PMID- 19560581 TI - Cash transfers for children--investing into the future. PMID- 19560575 TI - Intestinal barrier function: molecular regulation and disease pathogenesis. AB - The intestinal epithelium is a single-cell layer that constitutes the largest and most important barrier against the external environment. It acts as a selectively permeable barrier, permitting the absorption of nutrients, electrolytes, and water while maintaining an effective defense against intraluminal toxins, antigens, and enteric flora. The epithelium maintains its selective barrier function through the formation of complex protein-protein networks that mechanically link adjacent cells and seal the intercellular space. The protein networks connecting epithelial cells form 3 adhesive complexes: desmosomes, adherens junctions, and tight junctions. These complexes consist of transmembrane proteins that interact extracellularly with adjacent cells and intracellularly with adaptor proteins that link to the cytoskeleton. Over the past decade, there has been increasing recognition of an association between disrupted intestinal barrier function and the development of autoimmune and inflammatory diseases. In this review we summarize the evolving understanding of the molecular composition and regulation of intestinal barrier function. We discuss the interactions between innate and adaptive immunity and intestinal epithelial barrier function, as well as the effect of exogenous factors on intestinal barrier function. Finally, we summarize clinical and experimental evidence demonstrating intestinal epithelial barrier dysfunction as a major factor contributing to the predisposition to inflammatory diseases, including food allergy, inflammatory bowel diseases, and celiac disease. PMID- 19560582 TI - Moving forward with maternal health and human rights. PMID- 19560583 TI - Alcohol: a global health priority. PMID- 19560584 TI - Action needed to tackle a global drink problem. PMID- 19560585 TI - A case study in how harmful alcohol consumption can be. PMID- 19560586 TI - Primary immunodeficiency diseases: the J Project. PMID- 19560588 TI - Save Somalia! PMID- 19560587 TI - Judicialisation of the right to health in Brazil. PMID- 19560589 TI - Sally Casswell: champion for communities tackling alcohol. Interview by Kelly Morris. PMID- 19560590 TI - Global health and the Bill & Melinda Gates Foundation. PMID- 19560591 TI - Global health and the Bill & Melinda Gates Foundation. PMID- 19560592 TI - Global health and the Bill & Melinda Gates Foundation. PMID- 19560593 TI - Preserving objectivity in medical education. PMID- 19560594 TI - Preserving objectivity in medical education. PMID- 19560595 TI - Preserving objectivity in medical education. PMID- 19560597 TI - Preserving objectivity in medical education. PMID- 19560596 TI - Moxifloxacin versus ethambutol in initial tuberculosis treatment. PMID- 19560598 TI - Moxifloxacin versus ethambutol in initial tuberculosis treatment. PMID- 19560600 TI - CABG versus PCI for multivessel coronary artery disease. PMID- 19560602 TI - Alcohol and cause-specific mortality in Russia: a retrospective case-control study of 48,557 adult deaths. AB - BACKGROUND: Alcohol is an important determinant of the high and fluctuating adult mortality rates in Russia, but cause-specific detail is lacking. Our case-control study investigated the effects of alcohol consumption on male and female cause specific mortality. METHODS: In three Russian industrial cities with typical 1990s mortality patterns (Tomsk, Barnaul, Biysk), the addresses of 60,416 residents who had died at ages 15-74 years in 1990-2001 were visited in 2001-05. Family members were present for 50,066 decedents; for 48,557 (97%), the family gave proxy information on the decedents' past alcohol use and on potentially confounding factors. Cases (n=43,082) were those certified as dying from causes we judged beforehand might be substantially affected by alcohol or tobacco; controls were the other 5475 decedents. Case versus control relative risks (RRs; calculated as odds ratios by confounder-adjusted logistic regression) were calculated in ever-drinkers, defining the reference category by two criteria: usual weekly consumption always less than 0.5 half-litre bottles of vodka (or equivalent in total alcohol content) and maximum consumption of spirits in 1 day always less than 0.5 half-litre bottles. Other ever-drinkers were classified by usual weekly consumption into three categories: less than one, one to less than three, and three or more (mean 5.4 [SD 1.4]) bottles of vodka or equivalent. FINDINGS: In men, the three causes accounting for the most alcohol-associated deaths were accidents and violence (RR 5.94, 95% CI 5.35-6.59, in the highest consumption category), alcohol poisoning (21.68, 17.94-26.20), and acute ischaemic heart disease other than myocardial infarction (3.04, 2.73-3.39), which includes some misclassified alcohol poisoning. There were significant excesses of upper aerodigestive tract cancer (3.48, 2.84-4.27) and liver cancer (2.11, 1.64 2.70). Another five disease groups had RRs of more than 3.00 in the highest alcohol category: tuberculosis (4.14, 3.44-4.98), pneumonia (3.29, 2.83-3.83), liver disease (6.21, 5.16-7.47), pancreatic disease (6.69, 4.98-9.00), and ill specified conditions (7.74, 6.48-9.25). Although drinking was less common in women, the RRs associated with it were generally more extreme. After correction for reporting errors, alcohol-associated excesses accounted for 52% of all study deaths at ages 15-54 years (men 8182 [59%] of 13968, women 1565 [33%] of 4751) and 18% of those at 55-74 years (men 3944 [22%] of 17,536, women 1493 [12%] of 12 302). Allowance for under-representation of extreme drinkers would further increase alcohol-associated proportions. Large fluctuations in mortality from these ten strongly alcohol-associated causes were the main determinants of recent fluctuations in overall mortality in the study region and in Russia as a whole. INTERPRETATION: Alcohol-attributable mortality varies by year; in several recent years, alcohol was a cause of more than half of all Russian deaths at ages 15-54 years. Alcohol accounts for most of the large fluctuations in Russian mortality, and alcohol and tobacco account for the large difference in adult mortality between Russia and western Europe. FUNDING: UK Medical Research Council, Cancer Research UK, British Heart Foundation, International Agency for Research on Cancer, and European Commission Directorate-General for Research. PMID- 19560603 TI - Status vasovagalis. PMID- 19560604 TI - Global burden of disease and injury and economic cost attributable to alcohol use and alcohol-use disorders. AB - Alcohol consumption has been identified as an important risk factor for chronic disease and injury. In the first paper in this Series, we quantify the burden of mortality and disease attributable to alcohol, both globally and for ten large countries. We assess alcohol exposure and prevalence of alcohol-use disorders on the basis of reviews of published work. After identification of other major disease categories causally linked to alcohol, we estimate attributable fractions by sex, age, and WHO region. Additionally, we compare social costs of alcohol in selected countries. The net effect of alcohol consumption on health is detrimental, with an estimated 3.8% of all global deaths and 4.6% of global disability-adjusted life-years attributable to alcohol. Disease burden is closely related to average volume of alcohol consumption, and, for every unit of exposure, is strongest in poor people and in those who are marginalised from society. The costs associated with alcohol amount to more than 1% of the gross national product in high-income and middle-income countries, with the costs of social harm constituting a major proportion in addition to health costs. Overall, we conclude that alcohol consumption is one of the major avoidable risk factors, and actions to reduce burden and costs associated with alcohol should be urgently increased. PMID- 19560605 TI - Effectiveness and cost-effectiveness of policies and programmes to reduce the harm caused by alcohol. AB - This paper reviews the evidence for the effectiveness and cost-effectiveness of policies and programmes to reduce the harm caused by alcohol, in the areas of education and information, the health sector, community action, driving while under the influence of alcohol (drink-driving), availability, marketing, pricing, harm reduction, and illegally and informally produced alcohol. Systematic reviews and meta-analyses show that policies regulating the environment in which alcohol is marketed (particularly its price and availability) are effective in reducing alcohol-related harm. Enforced legislative measures to reduce drink-driving and individually directed interventions to already at-risk drinkers are also effective. However, school-based education does not reduce alcohol-related harm, although public information and education-type programmes have a role in providing information and in increasing attention and acceptance of alcohol on political and public agendas. Making alcohol more expensive and less available, and banning alcohol advertising, are highly cost-effective strategies to reduce harm. In settings with high amounts of unrecorded production and consumption, increasing the proportion of alcohol that is taxed could be a more effective pricing policy than a simple increase in tax. PMID- 19560606 TI - Reducing harm from alcohol: call to action. AB - Despite clear evidence of the major contribution alcohol makes to the global burden of disease and to substantial economic costs, focus on alcohol control is inadequate internationally and in most countries. Expansion of industrial production and marketing of alcohol is driving alcohol use to rise, both in emerging markets and in young people in mature alcohol markets. Cost-effective and affordable interventions to restrict harm exist, and are in urgent need of scaling up. Most countries do not have adequate policies in place. Factors impeding progress include a failure of political will, unhelpful participation of the alcohol industry in the policy process, and increasing difficulty in free trade environments to respond adequately at a national level. An effective national and international response will need not only governments, but also non governmental organisations to support and hold government agencies to account. International health policy, in the form of a Framework Convention on Alcohol Control, is needed to counterbalance the global conditions promoting alcohol related harm and to support and encourage national action. PMID- 19560607 TI - Recurrent pneumothoraces, rash, and polyuria. PMID- 19560608 TI - [Aquagenic palmoplantar keratoderma and cystic fibrosis]. PMID- 19560609 TI - [Novel adverse effects of laser-assisted axillary hair removal]. AB - BACKGROUND: Several adverse effects of depilatory laser may now be commonly expected in relation to skin type and anatomic location. We report and analyse unusual events in patients undergoing laser-assisted axillary hair removal, including hyperhidrosis, bromhidrosis and leukotrichia. OBJECTIVE: The aim of this study was to examine a large patient population, assess the frequency of these adverse effects, and establish a correlation with different hair-removal laser protocols. PATIENTS AND METHODS: A retrospective case-control study of patients undergoing laser-assisted axillary hair removal was conducted to determine the incidence of these adverse effects in relation to skin type and types of laser used. RESULTS: Hyperhidrosis, bromhidrosis and leukotrichia were seen in 11, 4, and 2% of patients respectively. Hyperhidrosis was significantly less frequent in patients with skin types III and IV than in those with skin type II or V. Combined diode and alexandrite laser sessions were associated with a significantly higher incidence of hyperhidrosis compared to diode or alexandrite sessions alone. Regarding bromhidrosis and leukotrichia, no significant correlation with age, skin type or laser settings was revealed by the statistical analysis. CONCLUSION: Hyperhidrosis, bromhidrosis and leukotrichia are likely new adverse effects of laser-assisted axillary hair removal. PMID- 19560610 TI - [Desmoplastic trichoepithelioma: a clinicopathological study, including a comparison with morpheiform basal cell carcinoma]. AB - BACKGROUND: Desmoplastic trichoepithelioma (DTE) is an uncommon form of adnexal tumour that was described for the first time as a separate clinicopathological entity in 1976. We carried out a retrospective histopathological study of a large series of cases of DTE in order to better characterise this tumour and compare it with sclerodermiform basal cell carcinoma (BCC), which is in fact the most common as well as the most complex type of differential diagnosis. PATIENTS AND METHODS: We included in this study all cases of DTE diagnosed between 1979 and 2001 at our dermatopathology laboratory. The clinical features were taken from the patient files. Diagnosis was confirmed by two different examiners and all microscopic elements were reviewed. The same clinical and demographic data were collected for cases of BCC diagnosed over the same period. RESULTS: We included 68 cases of DTE in 67 patients in our study, of whom 83.5% were women, and the mean age was 42.8 years. Lesions were found primarily on the head (98.5% of cases), mainly on the cheeks (29.2% of cases) and forehead (23.1% of cases). The diagnosis was only made by the clinician in four cases; in 38 cases the diagnosis made was BCC. In all cases, histological examination revealed thin lines of basaloid epithelial cells associated with small keratinising cysts. A common finding was granulomas with foreign bodies and calcifications. In six cases (8.8%), a tumour combining DTE with an intradermal naevus was observed. Mean clinical follow-up of 8 years (1 to 23 years) in 29 patients showed absence of relapse or metastasis. Over the same period, 662 cases of BCC were recorded in 499 patients, 58.9% of whom were women. The mean age was 65.6 years. The principal location was the nose (34% of cases). No cases of associated naevus were recorded. COMMENTS: DTE is a firm plaque-like lesion, flesh coloured or yellowish, and generally with a depressed centre; it is seen primarily in middle-aged women and occurs principally on the cheeks and forehead. There is a significant association with naevus, a singular feature among adnexal tumours. On average, it is 10 times less common than BCC. The size of our study groups shows for the first time distinguishing features with regard to BCC, for which the age of onset is far higher and predominance among women less marked. Further, the sites are different, with BCC being seen predominantly on the nose while DTE is seen mainly on the cheeks. A suggested clinical diagnosis is thus possible. PMID- 19560611 TI - [Multiple Bowen disease of the lower limbs in elderly women: a rare clinical subset involving therapeutic difficulties]. AB - BACKGROUND: Cutaneous Bowen's disease (CBD) is a form of intraepithelial squamous cell carcinoma that usually presents as a solitary lesion. We report four similar cases of a peculiar and well-delimited clinical subset of multiple Bowen's disease seen in the lower limbs in elderly women and associated with specific therapeutic problems. OBSERVATIONS: Four women aged over 70 years presented with multiple CBD limited to the lower limbs associated with squamous cell and superficial basal cell carcinomas along with actinic keratosis. No significant aetiological factors were present apart from chronic sun exposure other than one case possibly involving immunodeficiency. The four patients were treated using photodynamic therapy, and partial clinical response and good tolerance were observed. DISCUSSION: These four cases share numerous clinical similarities: elderly women, markers of chronic sun exposure, lack of other aetiological factors such as arsenic or irradiation, localization of the lesions (multiple and/or continuous layer pattern, restricted to the lower limbs in all cases) and a chronic course. The frequency of this subset is probably underestimated due to absence of biopsies or to inconclusive histology reports. Photodynamic therapy yields valuable results with a good efficacy/safety ratio compared to imiquimod or 5-fluorouracil. However, while this treatment could be considered a first-line option in multiple CBD, its therapeutic value requires more detailed evaluation. PMID- 19560612 TI - [Epidermolysis bullosa acquisita of childhood]. AB - BACKGROUND: Epidermolysis bullosa acquisita (EBA) is a subepidermal autoimmune blistering disease characterized immunologically by autoantibodies to type VII collagen. Its occurrence in childhood is rare. Thirty-five cases have been described to date in the literature. PATIENTS AND METHODS: We report the case of an 8-year-old girl presenting blistering lesions on the cheeks, extremities and limb extension areas. The diagnosis of EBA was confirmed by histology, direct immunofluorescence of a perilesional skin biopsy specimen, indirect immunofluorescence on salt-split skin substrate and direct electron microscopy. The patient was controlled clinically under treatment with dapsone alone. DISCUSSION: This 36th childhood case of EBA presented typical clinical features, a similar prognosis and comparable treatment response to other paediatric cases. Clinical presentation is inflammatory and affects the face. As in our case, in childhood, prognosis is often better than in adults without the need for immunosuppressive agents. PMID- 19560613 TI - [A pseudotumoral cutaneous form of sarcoidosis associated with non-Hodgkin lymphoma]. AB - BACKGROUND: Sarcoidosis may be revealed by a variety of cutaneous signs, occasionally atypical. It may also be associated with a lymphoma. Herein we report an original case of pseudotumoral cutaneous sarcoidosis associated with malignant non-Hodgkin's lymphoma (MNHL). PATIENTS AND METHODS: A 64-year-old man was hospitalised for follicular and diffuse large B-cell MNHL localised in the cervical, subclavicular and inguinal lymph nodes. The skin examination revealed a bulky, erythematous, purple, infiltrated mass in the right lumbar region and two similar but smaller plaques in the dorsal region. The lesions were asymptomatic, present for six years, and stable. Histological examination of a skin biopsy sample revealed an epithelioid giant-cell granuloma without caseous necrosis. Bacteriological and mycobacteriological samples were sterile. Mediastinal adenopathies and pulmonary micronodules were found on CAT scan. A diagnosis of cutaneous and pulmonary sarcoidosis associated with the lymphoma was made. Polychemotherapy using CHOP combined with rituximab resulted in remission of the lymphoma but was ineffective against the sarcoidosis. COMMENTS: Pseudotumoral forms of sarcoidosis are rare and have a misleading clinical aspect. In our case, the cutaneous mass was evocative of a secondary lymphoma site. The diagnosis of sarcoidosis was based on clinical and laboratory findings and after elimination of other potential causes of granuloma. The appearance of a lymphoma in a patient with a history of sarcoidosis is rare but not fortuitous, since the notion of "sarcoidosis-lymphoma syndrome" exists. The incidence of lymphomas is 5.5-fold higher in patients presenting sarcoidosis than among the general population. Their time to onset is around 7 years after the discovery of sarcoidosis. The most common forms involve Hodgkin's disease. The efficacy of therapy on the blood disease underlying the sarcoidosis is variable. PMID- 19560614 TI - [Malignant pyoderma with cranial osteolysis]. AB - BACKGROUND: Malignant pyoderma or pyoderma gangrenosum with cephalic involvement is a rare disease characterized by chronic aggressive local progression. We report a case of malignant pyoderma with cranial osteolysis. PATIENTS AND METHODS: A 42-year-old man had been presenting chronic ulcerative dermatosis involving the entire scalp since the age of sixteen. Clinical examination, histopathological analysis of the skin sample and negative aetiological tests suggested a diagnosis of cephalic pyoderma gangrenosum. Different treatments were ineffective. The patient was subsequently admitted with convulsions revealing cranial osteolysis. A cranial scan confirmed the osteolysis and histopathological analysis of the bone sample revealed non-specific suppurative osteitis. Treatment with corticosteroids and thalidomide was effective. Skin and bone grafting was proposed but was not carried out due to the chronic progression and absence of stable wound healing. DISCUSSION: Malignant pyoderma is a rare destructive, ulcerating dermatosis. It is characterized by a chronic course and resistance to treatment. Our case report is original in terms of the involvement of underlying cranial bone revealed by convulsions. This bone involvement makes treatment of the dermatosis more difficult. PMID- 19560615 TI - [Sacral chordoma: a rare tumor of the buttocks]. AB - INTRODUCTION: Described and recognized for the first time as a pathological entity by Virchow in 1857, chordoma is a tumour of embryonic origin secondary to an attack of the notochord. In most cases it is asymptomatic, resulting in fairly late diagnosis. PATIENTS AND METHODS: We report the case of a 62-year-old patient presenting a bulky tumefaction, nodular in places, not very painful, and extending towards the anal area, scrotum and the posterior aspect of the upper left thigh. Histopathological examination of a macrobiopsy sample of this tumefaction pointed to chordoma. On magnetic resonance imaging (MRI), the tumour presented multiple ramifications extending towards the scrotal area, the sciatic area and the posterior aspect of the left thigh. Palliative tumorectomy was performed. Given the very slow progression of the tumour and the risk of adverse effects in such a large tumoral exposure field, radiotherapy was ruled out. DISCUSSION: This is a typical observation of a rare tumour that dermatologists may encounter. PMID- 19560617 TI - [Urachal sinus presenting as umbilical drainage in an adult male]. AB - BACKGROUND: Although urachal remnants are very rare, dermatologists must be aware of them in order to ensure suitable treatment by urologists. Lack of appropriate treatment exposes patients to the risks of recurrent infectious complications or degeneration to vesical adenocarcinoma. CASE REPORT: A 32-year-old man consulted for mild and recurring umbilical weeping for approximately 1 year. He had previously undergone unsuccessful treatment, comprising application of liquid nitrogen and of silver nitrate to an umbilical lesion measuring several millimetres regarded as pyogenic granuloma. The diagnosis of urachal sinus was suspected clinically (umbilical weeping or inflammation) and confirmed by urological scan. DISCUSSION: A defect involving partial or total obliteration of the urachal canal after the fifth month of gestation can result in four types of benign abnormalities. Patent urachus (50%) is diagnosed during the neonatal period, while in adults, the most frequent form is a urachal cyst (30%), with sinuses (15%) and the diverticula (5%) being very rare. The urachus and its implantation base on the bladder must be removed surgically. Laparoscopic surgery offers a valuable alternative to open-surgery and probably leads to less morbidity and shorter convalescence in active young subjects. PMID- 19560616 TI - [Chloroquine cardiotoxicity in long-term lupus therapy in two patients]. AB - BACKGROUND: The antimalarial compounds chloroquine and hydroxychloroquine are widely used in the treatment of connective tissue diseases and are usually well tolerated. We report two cases of chloroquine cardiotoxicity. PATIENTS AND METHODS: Two women (aged 43 and 48 years) were treated for 5 years for lupus. They developed severe conduction disturbances requiring a pacemaker. Plasma chloroquine concentrations were abnormally high in both cases. In one case, a genetic polymorphism modulating the activity of a cytochrome involved in chloroquine metabolism (CYP2C8) was identified. DISCUSSION: Since 1965, 60 cases of occasionally severe cardiotoxicity have been reported following long-term treatment with chloroquine in most cases, but also with hydroxychloroquine. This toxicity must be detected early and close cardiac assessment is required. PMID- 19560618 TI - [Amoxicillin-induced phototoxicity]. PMID- 19560619 TI - [A case of indigenous skin infection caused by methicillin-resistant Staphylococcus aureus USA300 in France]. PMID- 19560620 TI - [Mixed facial dermatitis: a common disorder meriting separate treatment. A study of 25 cases]. PMID- 19560621 TI - [Feverish skin eruption]. PMID- 19560622 TI - [A necrotic calf lesion]. PMID- 19560623 TI - [Pseudoxanthoma elasticum in childhood]. PMID- 19560624 TI - [Immunologic contact urticaria from cosmetics]. PMID- 19560625 TI - [Vascular patterns of dermal nevus]. PMID- 19560626 TI - [Ichthyosis: a rich field for research]. PMID- 19560627 TI - [Linear lichen]. PMID- 19560628 TI - Relation between plasma brain-derived neurotrophic factor and nerve growth factor in the male patients with alcohol dependence. AB - Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) are thought to be related to neuroprotection in cell culture and animal studies. Our aim was to verify the changes in human plasma BDNF and NGF concentrations induced by chronic alcohol use. Forty-one male patients with alcohol dependence were sampled the next morning of admission and compared with 41 healthy male subjects. Plasma BDNF and NGF were assayed using an enzyme-linked immunosorbent assay (ELISA). Mean plasma BDNF level was significantly higher in the patients with alcohol dependence (3502.21+/-1726.9 pg/mL) compared with the healthy subjects (861.75+/-478.9 pg/mL) (P=.000). Mean plasma NGF level was also significantly higher in patients with alcohol dependence (137.64+/-32.7 pg/mL) than in healthy subjects (112.61+/-90.2 pg/mL) (P=.012). Plasma BDNF and NGF levels showed significant negative correlation in alcohol dependence group (r=-0.388, P=.012). Increased plasma BDNF and NGF with negative correlation in alcohol-dependent patients may have some role in the regeneration of damage done by chronic alcohol use. PMID- 19560629 TI - Ethanol increases desensitization of recombinant GluR-D AMPA receptor and TARP combinations. AB - Glutamate receptors are important target molecules of the acute effect of ethanol. We studied ethanol sensitivity of homomeric GluR-D receptors expressed in human embryonic kidney 293 cells and examined whether recently discovered transmembrane alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor regulatory proteins (TARPs) affect ethanol sensitivity. Coexpression of the TARPs, stargazin, and gamma4 increased the time constant (tau-value) of current decay in the presence of agonist, thus slowing the onset of desensitization and increasing the steady-state current. Ethanol produced less inhibition of the peak current than the steady-state current for all types of the GluR-D receptors. In addition, ethanol concentration-dependently accelerated the rate of desensitization, measured as the tau-value of fast decay of peak current. This effect was enhanced with coexpression of TARPs. The recovery from desensitization was slowed down by coexpression of gamma4 but ethanol did not affect this process in any GluR-D combination. The results support the idea that increased desensitization is an important mechanism in the ethanol inhibition of AMPA receptors and indicate that coexpression of TARPs can alter this effect of ethanol. PMID- 19560631 TI - Proceedings of the 2008 annual meeting of the Fetal Alcohol Spectrum Disorders Study Group. AB - The annual meeting of the Fetal Alcohol Spectrum Disorders Study Group (FASDSG) was held on June 28, 2008 in Washington DC, as a satellite to the Research Society on Alcoholism meeting. The FASDSG membership includes clinical, basic, and social scientists who meet to discuss recent advances and issues in FASD research. The main theme of the meeting was "Factors that Influence Brain and Behavioral Development: Implications for Prevention and Intervention." Two keynote speakers, Dr. Stephen Suomi and Dr. Carl Keen addressed how early environment and nutrition may influence outcome after prenatal alcohol exposure. The final keynote speaker, Kathy Mitchell, addressed issues regarding the relationship between scientists and the families with children with FASD. Members of the FASDSG provided updates on new findings through brief (FASt) data reports and national agency representatives provided updates of activities and funding priorities. Presentations were also made by recipients of the Student Research Merit award and Rosett award. PMID- 19560630 TI - Activation of MEK 1/2 and p42/44 MAPK by angiotensin II in hepatocyte nucleus and their potentiation by ethanol. AB - Hepato-subcellular effect of angiotensin II (Ang II) and ethanol on the p42/44 mitogen-activated protein kinase (MAPK) and MAPK kinase (MEK 1/2) was investigated in the nucleus of rat hepatocytes. Hepatocytes were treated with ethanol (100 mM) for 24h and stimulated with Ang II (100 nM, 5 min). The levels of p42/44 MAPK and MEK 1/2 were monitored in the nuclear fraction using antibodies. Ang II itself caused significant accumulation of phosphorylated p42/44 MAPK (phospho-p42/44 MAPK) in the nucleus without any significant translocation of p42/44 MAPK protein thereby suggesting activation of p42/44 MAPK in the nucleus. Ang II caused marked accumulation of phosphorylated MEK 1/2 (phospho-MEK 1/2) in the nucleus without any significant accumulation of MEK 1/2 protein. Ratio of phospho-MEK 1/2 to MEK 1/2 protein in the nucleus after Ang II treatment was 2.4 times greater than control suggesting phosphorylation of MEK 1/2 inside the nucleus. Ethanol had no effect on the protein level or the activation of p42/44 MAPK in the nucleus. Ethanol treatment potentiated nuclear activation of p42/44 MAPK by Ang II but not translocation of p42/44 MAPK protein. This was accompanied by potentiation of Ang II-stimulated accumulation of phospho MEK 1/2 in the nucleus by ethanol. MEK 1/2 inhibitor, U-0126 inhibited Ang II response and its potentiation by ethanol. These results suggest that Ang II mediated accumulation of phospho-p42/44 MAPK in the hepatocyte nucleus involves MEK 1/2-dependent activation and this effect is potentiated by ethanol. PMID- 19560632 TI - Let's go to the video. PMID- 19560633 TI - Local anesthetics induce chondrocyte death in bovine articular cartilage disks in a dose- and duration-dependent manner. AB - PURPOSE: The purpose of this study was to evaluate the effect of various local anesthetics on chondrocyte viability in articular cartilage by use of a bovine disk model. METHODS: Full-thickness bovine cartilage disks were isolated from the condylar surfaces of the radial-carpal joint by use of a 4-mm biopsy punch and were incubated in various concentrations of local anesthetics (e.g., bupivacaine) for varying amounts of time and stained for membrane integrity by use of ethidium bromide and SYTO 13 stain (Molecular Probes, Carlsbad, CA). Cell and nuclear morphology was assessed by transmission electron microscopy. RESULTS: The addition of local anesthetics (i.e., 0.25% bupivacaine, 1% lidocaine, and 0.5% ropivacaine) to bovine articular cartilage disks had a negative effect on chondrocyte viability. Culturing bovine articular cartilage disks for increasing periods of time decreased chondrocyte viability for each of the local anesthetics, with significant negative correlations being shown between time of exposure to the drug and chondrocyte viability. These effects were also affected by the presence or absence of epinephrine in local anesthetic preparations. CONCLUSIONS: Our results suggest that local anesthetics (i.e., bupivacaine, lidocaine, or ropivacaine) can have a detrimental effect on chondrocyte viability in bovine articular cartilage disks in a dose- and duration-dependent manner. CLINICAL RELEVANCE: After arthroscopic surgery, it has been common practice to inject various local anesthetics into the joint for pain relief. Because adult chondrocytes have little or no capacity to regenerate, these results suggest that high-dose, long-term intra-articular administration of local anesthetics should be performed with caution. PMID- 19560634 TI - Arthroscopic treatment for pigmented villonodular synovitis of the shoulder associated with massive rotator cuff tear. AB - PURPOSE: Our purpose was to investigate arthroscopic treatment of patients diagnosed with pigmented villonodular synovitis (PVNS) of the shoulder and massive rotator cuff tear with the initial presentation of large, recurrent joint effusion. METHODS: From December 2005 to June 2007, 5 patients (3 males and 2 females) diagnosed with PVNS of the shoulder and massive rotator cuff tear were treated with arthroscopic synovectomy, partial cuff repair, or debridement if the cuff was irreparable. All 5 patients were followed-up for a mean of 22.4 months (range, 12 to 33 months). Outcomes were measured with use of the American Shoulder and Elbow Surgeons (ASES) and University of California at Los Angeles (UCLA) scoring systems. Two patients received partial rotator cuff repair by suture anchors and another 2 received suture repairs only. All of the patients had residual tear with variable sizes. RESULTS: With a mean follow-up of 22.4 months (range, 12 to 33 months), the mean ASES and UCLA scores improved from preoperative values of 48.2 and 7.8 to 80.0 and 29.6 points, respectively (P < .05). All patients were satisfied with the procedure, and no signs of recurrence were noted during the follow-up period. CONCLUSIONS: Five cases of PVNS of the shoulder and massive rotator cuff tears with the initial symptoms of shoulder effusion and function limitation were reported. After arthroscopic synovectomy and partial rotator cuff repair or debridement, all patients gained symptomatic and limited functional improvement at an average follow-up of 22 months. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 19560635 TI - Meniscal allograft size can be predicted by height, weight, and gender. AB - PURPOSE: Our purpose was to determine if height, weight, and gender can be used to accurately predict proper meniscal allograft dimensions. METHODS: Data were obtained from the Joint Restoration Foundation (AlloSource, Centennial, CO) regarding meniscal size and patient characteristics from meniscal donors. Donor height, weight, sex, age, and anatomic meniscal dimensions were recorded for 930 donor menisci in 664 patients. Multivariate regressions were completed using gender, height, and weight as independent variables and lateral meniscus length, lateral meniscus width, medial meniscus length, and medial meniscus width as dependent variables. The regression formulas were then reapplied to the data in order to produce estimated meniscus dimensions based on donor height, weight, and gender. A 90:10 split of the data was used to validate the regression models. Predicted meniscal size was then compared to actual meniscal size and the results compared to current measurement techniques. RESULTS: Regression formulas showed the ability to predict meniscal size based on gender, height, and weight with standard deviations (SDs) equal to or less than current radiographic techniques (SD, 6.4% to 8.2%). Average differences between predicted size and actual size ranged from 5.2% to 6.5% for length and 5.2% to 6.0% for width. Patient height was found to be a much more powerful predictor of meniscal size than patient weight. Data from the 90:10 split of data validated the model on an independent sample. These validated outputs were then compared to contemporary techniques and found to have lower SDs and average error rates in the majority of cases. CONCLUSIONS: We have proposed a validated regression model that uses height, weight, and gender variables to accurately predict required allograft meniscal size. We compared it against previously published data for radiographic and magnetic resonance imaging sizing techniques and found it to produce results that were, overall, slightly more accurate. CLINICAL RELEVANCE: This model provides a novel method for sizing meniscal allografts. PMID- 19560636 TI - Arthroscopic identification of isolated tear of the posterolateral bundle of the anterior cruciate ligament. AB - PURPOSE: The goal of this study was to arthroscopically identify and assess the progressive changes in isolated ruptures of the posterolateral bundle of the anterior cruciate ligament (ACL) over time. METHODS: This prospective study investigated 174 patients for isolated posterolateral bundle tears during arthroscopic ACL reconstruction. The preoperative side-to-side anterior laxity was measured in all patients. The torn ACLs were inspected and analyzed arthroscopically to determine the tear pattern. RESULTS: Complete ACL tears were identified in 78.7% of patients and partial ACL tears in 21.3%. Isolated anteromedial bundle tears were identified in 22 patients and posterolateral bundle tears in 15 patients. Statistical analysis showed a significant difference in preoperative differential knee laxity between the group with complete ACL rupture and the group with partial ACL rupture. On arthroscopic evaluation, the posterolateral bundle had retracted distally toward the tibial surface over time. The amount of retraction was correlated to the time period from injury to reconstruction. CONCLUSIONS: In this study the exact diagnosis of an ACL partial tear was made at arthroscopy in all cases. Our observations confirm the evolution of the ruptured posterolateral bundle, which shows a retraction toward the tibia over time. LEVEL OF EVIDENCE: Level II, development of diagnostic criteria based on consecutive patients with a universally applied gold standard. PMID- 19560637 TI - In vivo magnetic resonance imaging measurement of tibiofemoral relation with different knee flexion angles after single- and double-bundle anterior cruciate ligament reconstructions. AB - PURPOSE: Double-bundle anterior cruciate ligament (ACL) reconstruction has been shown to restore better kinematics in vitro, but it is uncertain whether this technique can achieve this in vivo. We investigated whether anatomic double bundle ACL reconstruction can restore a better tibiofemoral relation in the sagittal plane under static unloading conditions. METHODS: The tibiofemoral relation was assessed with an open magnetic resonance imaging scanner (0.5 T) in 15 patients with anatomic double-bundle reconstruction (double-bundle group) and 14 patients with single-bundle reconstruction (single-bundle group) by use of hamstring tendons. T1-weighted magnetic resonance imaging for both knees was obtained at 0 degrees, 45 degrees, 90 degrees, and 120 degrees of flexion without external force and muscle contraction 6 months after surgery. The position of the posterior femoral condyles relative to the tibia was measured in the midmedial and midlateral sagittal sections of the knee. Clinical evaluation (range of motion, KT-2000 measurement [MEDmetric, San Diego, CA], and pivot-shift test) was performed at the same time. RESULTS: KT-2000 testing showed that the mean side-to side difference in the double-bundle group (0.7 mm) was significantly smaller than that in the single-bundle group (1.7 mm). In the double-bundle group, the tibiofemoral relation in operated knees was not significantly different from that in the contralateral knees at 0 degrees, 45 degrees, 90 degrees, and 120 degrees of flexion, although at 0 degrees of flexion, the femoral condyles were positioned anteriorly relative to the tibia compared with that in the contralateral knees. Similar results were obtained in the single-bundle group. The difference in the tibiofemoral relation between reconstructed and contralateral knees was not significantly different between the 2 groups. CONCLUSIONS: According to KT-2000 measurement, this study found that anteroposterior stability was better with anatomic double-bundle ACL reconstruction than with single-bundle ACL reconstruction. However, under static conditions without an anterior drawer force, anatomic double-bundle ACL reconstruction did not show superiority in terms of restoring a better tibiofemoral relation compared with single-bundle ACL reconstruction. LEVEL OF EVIDENCE: Level III, comparative study. PMID- 19560638 TI - The effect of anterior cruciate ligament reconstruction on stride-to-stride variability. AB - PURPOSE: The purpose of our study was to investigate the functional outcome after anterior cruciate ligament (ACL) reconstruction using bone-patellar tendon-bone (BPTB) and quadrupled semitendinosus and gracilis tendon (ST/G) autografts by evaluating stride-to-stride variability. METHODS: Six patients with BPTB and 6 patients with STG ACL reconstruction, 2 years postoperatively, and 6 healthy control subjects walked on a treadmill at a self-selected pace while 2 minutes of continuous kinematic data were recorded with a 6-camera optoelectronic system. Stride-to-stride variability was calculated from the knee flexion/extension data using the nonlinear measure of approximate entropy, which estimates the regularity of movement patterns over time. RESULTS: ACL reconstruction affects stride-to-stride variability. Both the BPTB and the ST/G groups had significantly larger approximate entropy values than the healthy controls. No differences were found between the BPTB and the ST/G approximate entropy values. CONCLUSIONS: After ACL reconstruction using either BPTB or quadrupled ST/G, there is increased gait variability as compared to healthy individuals. This could be caused by the altered neuromuscular activity found in ACL-reconstructed limbs. LEVEL OF EVIDENCE: Level III, case control study. PMID- 19560639 TI - Anterior cruciate ligament reconstruction with bone-patellar tendon-bone autograft versus allograft. AB - PURPOSE: The purpose of this study was to analyze the clinical outcome of arthroscopic anterior cruciate ligament (ACL) reconstruction with bone-patellar tendon-bone autograft versus allograft. METHODS: Between May 2000 and June 2004, 172 patients undergoing arthroscopic bone-patellar tendon-bone ACL reconstruction were prospectively randomized into autograft (n = 86) or allograft (n = 86) groups. The senior surgeon performed all operations using the same surgical technique. Each fixation was performed by means of an interference screw. Patients were evaluated preoperatively and postoperatively at follow-up. Of the patients, 156 (76 in the autograft group and 80 in the allograft group) were available for full evaluation. Evaluations included a detailed history, physical examination, functional knee ligament testing, KT-2000 arthrometer testing (MEDmetric, San Diego, CA), Harner's vertical jump and Daniel's 1-leg hop tests, Lysholm score, Tegner score, International Knee Documentation Committee standard evaluation form, Cincinnati knee score, and radiograph. RESULTS: Demographic data were comparable between groups. The mean follow-up was 5.6 years for both groups. There were no statistically significant differences according to evaluations of outcome between the 2 groups except that patients in the allograft group had a shorter operation time and longer fever time postoperatively compared with the autograft group. The postoperative infection rates were 0% and 1.25% for the autograft group and allograft group, respectively. There was a significant difference (P < .05) in the development of osteoarthritis between the operated knee in comparison to the contralateral knee according to radiographs. However, no significant difference was found between the 2 groups at the final follow-up examination (P > .05). CONCLUSIONS: Both groups of patients achieved almost the same satisfactory outcomes after a mean of 5.6 years of follow-up. Allograft is a reasonable alternative to autograft for ACL reconstruction. LEVEL OF EVIDENCE: Level II, prospective comparative study. PMID- 19560640 TI - Magnetic resonance imaging of 3-dimensional in vivo tibiofemoral kinematics in anterior cruciate ligament-reconstructed knees. AB - PURPOSE: The purpose of this study was to use magnetic resonance imaging (MRI) to determine 3-dimensional knee kinematics after anterior cruciate ligament (ACL) reconstruction. METHODS: Nine ACL-reconstructed and contralateral knees were tested 12 +/- 8 months after surgery. MRI was performed at full extension and 40 degrees of knee flexion under simulated weight-bearing conditions. Femoral condyle positions, tibial rotation, contact area, and contact location were analyzed by use of MRI-based 3-dimensional models. RESULTS: When knees were fully extended, tibiae in ACL-reconstructed knees were externally rotated by 3.6 degrees +/- 4.2 degrees compared with contralateral knees. The external rotation was due to anterior subluxation of the medial side of the tibia. At 40 degrees of knee flexion, tibiae in ACL-reconstructed knees and contralateral knees were both internally rotated by 5.3 degrees. There were no significant differences in contact area or contact location between ACL-reconstructed and contralateral knees. When moving from extension to flexion, ACL-reconstructed knees exhibited 3.5 degrees +/- 5.9 degrees more internal tibial rotation than contralateral knees. CONCLUSIONS: Reconstruction of the ACL restored normal motion on the lateral side of the knee but not on the medial side, resulting in increased internal tibial rotation when moving from full extension to 40 degrees of flexion. These results suggest that ACL reconstruction does not restore normal kinematics on the medial side of the knee, which may lead to early cartilage degeneration. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 19560642 TI - Double-bundle posterior cruciate ligament reconstruction using a non-hardware suspension fixation technique and 8 strands of autogenous hamstring tendons. AB - PURPOSE: The purpose of this study was to evaluate the results of double-bundle posterior cruciate ligament (PCL) reconstruction using a non-hardware suspension fixation technique and 8 strands of autogenous hamstring tendons. METHODS: Twenty two cases of isolated chronic PCL rupture were reconstructed arthroscopically with 8 strands of autogenous hamstring tendons in a double-bundle and 4-tunnel manner. Quadruple-stranded semitendinosus tendon graft and quadruple-stranded gracilis tendon graft were used to reconstruct the anterolateral bundle and posteromedial bundle, respectively. The grafts were fixed by use of a non hardware suspension fixation technique. RESULTS: Nineteen patients were followed up for a minimum of 2 years. Before surgery, 15 patients had a 2+ posterior drawer test (PDT) and 4 patients had a 3+ PDT. At a minimum of 2 years after surgery, 17 patients (89.5%) had a negative PDT, 1 patient (5.3%) had a 1+ PDT, and 1 patient (5.3%) had a 2+ PDT. The mean KT-1000 examination results (MEDmetric, San Diego, CA) changed from 9.4 +/- 1.8 mm preoperatively to 1.0 +/- 1.0 mm postoperatively (P < .001). The stress radiography results changed from 10.6 +/- 2.0 mm preoperatively to 2.0 +/- 1.2 mm postoperatively (P < .001). According to the International Knee Documentation Committee knee examination form, the results were graded as normal in 15 patients (78.9%), nearly normal in 3 patients (15.8%), and abnormal in 1 patient (5.3%). The International Knee Documentation Committee subjective results increased from 65.6 +/- 5.1 to 92.1 +/ 3.7 (P < .001), and the Lysholm score increased from 63.5 +/- 4.9 to 92.5 +/- 4.1 (P < .001). The Tegner score was 5.1 before surgery and 6.3 at the last follow-up on average. CONCLUSIONS: The results of this study showed that arthroscopic double-bundle PCL reconstruction by use of a non-hardware suspension suture fixation technique and 8 strands of autogenous hamstring tendons can yield normal results in 78.9% of patients and nearly normal results in 15.8% at a minimum of 2 years. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 19560641 TI - Suspensory fixation of grafts in anterior cruciate ligament reconstruction: a biomechanical comparison of 3 implants. AB - PURPOSE: Our purpose was to evaluate the biomechanical difference between 2 new soft-tissue anterior cruciate ligament (ACL) fixation devices (RetroButton [Arthrex, Naples, FL] and ToggleLoc [Biomet, Warsaw, IN]) and a clinically established implant (EndoButton CL; Smith & Nephew, Andover, MA). METHODS: In test session 1 biomechanical testing was performed by use of 48 porcine femora fixed to a material testing machine. In session 2 the implants were tested alone. Cyclic loading was carried out for 1,000 cycles, followed by a load-to-failure test. The tested devices were as follows: EndoButton CL, 20-mm and 40-mm loops; RetroButton, 12/20-mm and 12/40-mm loops; and ToggleLoc, 20-mm and 40-mm loops. RESULTS: In test session 1 the EndoButton CL showed higher motion per cycle (0.30 +/- 0.09 mm for 20-mm loop and 0.43 +/- 0.03 mm for 40-mm loop) but lower plastic displacement values (0.66 +/- 0.19 mm for 20-mm loop and 0.50 +/- 0.13 mm for 40 mm loop) than the objects of comparison. The RetroButton devices were the stiffest (331.47 +/- 133.92 N/mm for 12/20-mm loop and 265.66 +/- 103.97 N/mm for 12/40-mm loop). The highest ultimate failure load, however, was shown by the EndoButton CL devices. In test session 2 the EndoButton CL oscillated the most. The ToggleLoc devices elongated more than the objects of comparison (0.66 +/- 0.12 mm for 20-mm loop and 0.76 +/- 0.06 mm for 40-mm loop). The RetroButton devices were both the stiffest among the respective implants (542.7 +/- 148.0 N/mm for 12/20-mm loop and 379.0 +/- 40.1 N/mm for 12/40-mm loop). The EndoButton CL showed the highest values for displacement to failure (3.6 +/- 0.4 mm for 20 mm loop and 6.4 +/- 0.4 mm for 40-mm loop). The ToggleLoc devices failed the latest on load-to-failure testing. CONCLUSIONS: All tested implants could provide adequate fixation strength. Despite advantages in the design of the EndoButton CL regarding its handling, the 2 newly released products showed superior material properties. CLINICAL RELEVANCE: Suspensory fixation of hamstring grafts in ACL reconstruction is frequently associated with bone tunnel enlargement. Material properties and implant design may limit graft-tunnel motion and result in enlargement of the femoral bone tunnel. With improved implant design, suspensory graft fixation may still be an attractive fixation technique in primary and revision ACL reconstruction. PMID- 19560643 TI - Arthroscopic subtalar fusion for post-traumatic subtalar arthritis. AB - PURPOSE: The purpose of this study was to determine the results of arthroscopic subtalar posterior facet fusion for subtalar arthritis after calcaneal fracture. METHODS: This is a retrospective case series on 10 patients (8 men and 2 women) who had isolated subtalar arthritis after calcaneal fracture. All patients were treated conservatively, but pain was not controlled. Only patients without hardware in the calcaneus were chosen for inclusion in the study. Patients were treated by posterior subtalar arthroscopic fusion. Fixation was done by percutaneous cannulated screws. Bone graft was not needed. All patients were evaluated preoperatively and postoperatively by the American Orthopaedic Foot & Ankle Society scoring system for the ankle and hind foot. The mean follow-up period was 28.4 months; only 1 case was missed during follow-up. RESULTS: There was statistically significant improvement in total American Orthopaedic Foot & Ankle Society score after 2 years' follow-up (P < .05). There was statistically significant improvement in pain and walking distance after follow-up (P < .05). The mean time for fusion was 11.44 weeks. Only 1 case had painful neuroma at the site of the anterolateral portal. CONCLUSIONS: Arthroscopically assisted posterior facet fusion of the subtalar joint is an effective method for management of subtalar arthritis after calcaneal fracture. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 19560644 TI - Bioabsorbable anchors in glenohumeral shoulder surgery. AB - The use of implants to provide glenohumeral soft tissue fixation has changed dramatically over the past few decades, from point tack fixation to metallic suture anchors to bioabsorbable suture anchors. Bioabsorbable suture anchors have largely replaced metallic anchors because of concerns of implant loosening, migration, and chondral injury. Although the safety and efficacy of bioabsorbable anchors has been well documented, there are numerous reports regarding the early failure related to implant bioabsorbable implant breakage or premature degradation. Patients with anchor-related complications generally present with pain and/or stiffness, and the surgeon should have a high index of suspicion if a patient does not progress as expected. Glenohumeral synovitis, glenoid osteolysis, loose bodies, and chondral injury are some of the notable complications that have been reported. Careful attention to proper anchor insertion techniques can limit the potential for complications. Newer materials, such as polyetheretherketone and other composites, have recently been introduced. These materials may address concerns of biocompatibility and material strength, but additional rigorous in vitro and in vivo trials need to be conducted before their use becomes widespread. PMID- 19560645 TI - Can the private practice of orthopaedic surgery survive the 21st century? PMID- 19560646 TI - Posterolateral sling reconstruction of the popliteus tendon: an all-arthroscopic technique. AB - Injuries to the posterolateral corner of the knee present with variable injury patterns that have produced a number of reconstructive procedures in the literature. We present an all-arthroscopic technique that reconstructs the popliteus tendon using either a semitendinosus autograft or an anterior tibialis allograft. After exposure of the posterior tibia by use of the posteromedial and trans-septal portals for visualization and posterolateral portal as a working portal, the popliteus musculotendinous junction is identified. In preparation for tibial tunnel drilling, a Kirschner wire is passed from the Gerdy tubercle to the popliteus musculotendinous junction on the posterior tibia, which is localized by use of an anterior cruciate ligament tibial tunnel guide brought in through the posterolateral portal. The 6-mm tibial tunnel is then created. The femoral insertion site is identified by use of the anterolateral portals for visualization and by use of the accessory superolateral portal for debriding the synovial fold at the insertion of the popliteus tendon. A K-wire is inserted, and a socket is established. The selected graft is then implanted by passing the graft through the tibial tunnel from anterior to posterior with a passing suture and pulling it up anteriorly and superiorly to the femoral socket. The graft is tensioned with the knee in 90 degrees of flexion and in neutral rotation before fixation of both ends of the graft with bio-interference screws. PMID- 19560647 TI - Arthroscopically assisted transfibular talar dome fixation with a headless screw. AB - We describe an articular talar dome fracture treated with the use of an arthroscopically assisted placement of a transfibular Acutrak screw (Accumed, Hillsboro, OR). This minimally invasive technique can be used to avoid the large incision used in the traditional open reduction and internal fixation of these fractures. Accurate reduction and stable fixation of fractures of the articular surface of the talus is a must, and this technique allows this while avoiding the morbidity of the open approach. PMID- 19560648 TI - Delayed incorporation of a TruFit plug: perseverance is recommended. AB - TruFit plugs (Smith & Nephew, Andover, MA) are synthetic polymer scaffolds that are inserted into an articular surface to provide a stable scaffold to encourage the regeneration of a full thickness of articular cartilage to repair chondral defects. Our unit has shown promising early results for the repair of small articular cartilage defects within the knee. Other series have reported "failures" in which patients have complained of persistent symptoms and joint effusion at 6 months after plug insertion and arthroplasty has been undertaken. We report a case of delayed incorporation of an articular cartilage defect of the lateral femoral condyle treated with 3 TruFit plugs. The patient eventually reported symptom alleviation and resumption of functional activity after 24 months of continued rehabilitation. We recommend that patients with continued symptoms persevere with rehabilitation and allow the regenerating articular cartilage time to mature fully before considering undertaking irreversible arthroplasty procedures. PMID- 19560649 TI - Bone graft substitute plug failure with giant cell reaction in the treatment of osteochondral lesions of the distal femur: a report of 2 cases with operative revision. AB - Although the treatment of symptomatic osteochondral lesions remains the focus of many orthopaedic clinicians, controversy remains regarding treatment choices and indications. Recently, the use of synthetic bone graft substitute composite plugs has been described to treat bone void lesions. We report 2 cases in which bone graft substitute plugs were used to graft bone defects in the knee and subsequently failed, requiring revision surgery. At revision arthroscopy, failure of incorporation was noted with foreign-body giant cells on histologic evaluation. Our technique for site debridement, revision bone grafting, and osteochondral resurfacing is presented. PMID- 19560650 TI - Arthroscopic hip surgery: Who can? Who should? PMID- 19560653 TI - Color pixels and grey speckles: new tools for intraoperative evaluation of left ventricular function. PMID- 19560654 TI - Echocardiographic imaging in clinical trials: American Society of Echocardiography Standards for echocardiography core laboratories: endorsed by the American College of Cardiology Foundation. PMID- 19560656 TI - Outcomes research: importance to (and of) echocardiography. PMID- 19560655 TI - Outcomes research in cardiovascular imaging: report of a workshop sponsored by the National Heart, Lung, and Blood Institute. AB - In July of 2008, the National Heart, Lung, and Blood Institute convened experts in noninvasive cardiovascular imaging, outcomes research, statistics, and clinical trials to develop recommendations for future randomized controlled trials of the use of imaging in: 1) screening the asymptomatic patient for coronary artery disease; 2) assessment of patients with stable angina; 3) identification of acute coronary syndromes in the emergency room; and 4) assessment of heart failure patients with chronic coronary artery disease with reduced left ventricular ejection fraction. This study highlights several possible trial designs for each clinical situation. PMID- 19560657 TI - Assessment of the right ventricle by echocardiography: a primer for cardiac sonographers. AB - The assessment of right ventricular (RV) structure and function by echocardiography has been improved by advancements that include Doppler tissue imaging, strain imaging, and three-dimensional imaging. Doppler tissue imaging and strain imaging can be useful for the assessment of regional RV systolic and diastolic function. Three-dimensional imaging has been reported to determine RV volumes and ejection fraction, which have previously been cumbersome to measure with conventional two-dimensional echocardiography. This article addresses the role of conventional and newer methods of echocardiography to assist sonographers in understanding the technical considerations, limitations, and pitfalls of image acquisition and analysis of RV structure and function. PMID- 19560658 TI - Two years of appropriateness criteria for echocardiography: what have we learned and what else do we need to do? PMID- 19560659 TI - Endurance exercise and the heart: multiple benefits but many unanswered questions. PMID- 19560660 TI - Stepping away from ritual right heart catheterization into the era of noninvasively measured pulmonary artery pressure. PMID- 19560661 TI - Mechanical dyssynchrony assessed by speckle tracking imaging as a reliable predictor of acute and chronic response to cardiac resynchronization therapy. AB - Speckle tracking echocardiography (STE) has the potential to detect the heterogeneous initiation of active myocardial contraction, which is the primary cause of left ventricular (LV) systolic dysfunction in patients with mechanical dyssynchrony. This study was designed to assess the ability to predict response to cardiac resynchronization therapy (CRT) of STE-derived dyssynchrony parameters in comparison with invasive hemodynamic assessments. Thirty patients referred for CRT were studied. The time difference of first peak (Td-(first peak)) and the maximum peak (Td-(max peak)) on the radial strain-time curves of the earliest and the latest segments were measured as the dyssynchrony parameter. Peak positive dP/dt (dP/dt(max)) was measured as the indicator of global LV systolic performance. CRT responders were defined as patients with LV end-systolic volume reduction greater than 15% at 3 months after CRT. CRT increased the dP/dt(max) compared with the baseline study (P < .001). Percent changes in the dP/dt(max) (dP/dt(max)) were significantly correlated with Td-(first peak) (R = 0.743, P < .001), but weakly correlated with Td-(max peak) (R = 0.390, P = .03). Twenty patients (66%) were identified as chronic CRT responders. Receiver operating characteristics analysis revealed that Td-(first peak) shared a similar ability with dP/dt(max) to detect chronic responders (Td-(first peak) >167.0 ms, area under the curve [AUC] 0.945; dP/dt(max) >16.2%, AUC 0.934) compared with Td-(max peak) (>194.5 ms, AUC 0.820). STE-derived Td-(first peak) showed a reliable ability to predict the acute and chronic response to CRT as well as dP/dt(max). PMID- 19560663 TI - Are we on the right way to calculate tau? PMID- 19560662 TI - Correlation between left ventricular end-diastolic pressure and peak left atrial wall strain during left ventricular systole. AB - OBJECTIVE: Left atrial (LA) reservoir function is determined by integration of LA relaxation and left ventricular (LV) systolic function, and LV diastolic dysfunction increases LA volume at end systole. This study investigates the effect of LV end-diastolic pressure on LA wall tension during LV systole. METHODS: A total of 101 stable patients with sinus rhythm undergoing cardiac catheterization were studied. LA wall extension during LV systole was evaluated as LA wall strain in the longitudinal direction obtained using two-dimensional ultrasound speckle tracking imaging. LV end-diastolic pressure and LV end systolic and end-diastolic volumes were obtained in cardiac catheterization, and LV ejection fraction was determined. RESULTS: Peak LA wall strain during LV systole had a significant inverse correlation with LV end-diastolic pressure (r = - 0.76, P < .0001). This correlation was also significant in patients with preserved LV systolic function (LV ejection fraction > or =50%) (r = - 0.64, P < .0001). In patients with peak LA wall strain during LV systole of less than 30%, 89% had elevated LV end-diastolic pressure (> or =16 mm Hg). CONCLUSION: Elevated LV end-diastolic pressure is associated with a decrease of peak LA wall strain in the longitudinal direction during LV systole. In patients with peak LA wall strain during LV systole of less than 30%, the majority had elevated LV end diastolic pressure, while most patients with peak LA wall strain during LV systole 45% or higher had normal LV end-diastolic pressures. In patients whose LV ejection fraction is 50% or more, when peak LA wall strain during LV systole is between 30% and 44%, it is not possible to predict LV end-diastolic pressure from peak LA wall strain measures. PMID- 19560665 TI - Fatal freely mobile left atrial thrombus: fallout of anticoagulation? AB - Rheumatic heart disease with moderate mitral stenosis and a large left atrial (LA) thrombus was diagnosed in a 45-year-old woman at a peripheral hospital. Oral anticoagulation was administered, and the patient was referred for further management. During the intervening period, sudden worsening of dyspnea developed in the patient and a repeat echocardiogram revealed a large thrombus freely vacillating within the left atrium. Urgent open mitral valvotomy and LA thrombus removal were planned. However, as the patient was being prepared for surgery, she had a fatal cardiorespiratory arrest and could not be revived. Warfarin is known to influence thrombus lysis, and it is possible that it may have severed the LA thrombus from the posterior LA wall by partially lysing it, leading to the formation of a ball-valve thrombus, which is a well-known risk factor for sudden death. PMID- 19560666 TI - Multimodal sonographic approach for assessing left ventricular outflow tract gradient. AB - Real-time three-dimensional echocardiography has now become an additional tool in the armamentarium of the perioperative echocardiographer. The use of all sonographic tools currently at the disposal of modern ultrasound technology enabled the correct diagnosis to be made in our patient. As a consequence, additional unnecessary surgery was avoided. PMID- 19560667 TI - Skin and soft tissue infections. Introduction. PMID- 19560668 TI - Future pharmacoeconomic criteria for the treatment of infections. AB - Skin and soft tissue infections (SSTI) are a common cause of antibiotic prescriptions and loss of time from school and work. Economic pressures on the delivery of healthcare have resulted in a focus on reducing resource utilization and costs, but not always on maintaining or improving the quality of patient care. The impact of treatment failure on the overall cost of therapy is not well reported for SSTI; however, older antibiotics, which can be perceived to be less expensive, may have a higher risk of treatment failure due to the development of resistance. This can increase overall direct and indirect treatment costs, including more office or emergency department visits for additional examinations/procedures, additional prescriptions for adjuvant therapies and supplementary antibiotics and, for some infections, hospitalization or increased length of hospital stay. Selection of antibiotics with an appropriate spectrum of activity is key, while the use of antibiotics with short and/or simple dosing regimens and a good tolerability profile, such as linezolid, which maximize patient adherence to the dosing regimen, can also play a role in ensuring that adequate antibiotic therapy is received. A further consideration is the time to symptom resolution, which can have a substantial impact on the patient's quality of life as well as affecting the indirect costs of the disease (e.g. loss of productivity and lost working days for the patient or parent). Thus, careful selection of antibiotic therapy, taking into account potential health outcomes and pharmacoeconomic benefits, is important to optimize clinical and bacteriological outcomes and thereby reduce overall costs to both the healthcare system and society. PMID- 19560669 TI - Why is community-associated MRSA spreading across the world and how will it change clinical practice? AB - Meticillin-resistant Staphylococcus aureus (MRSA) emerged in 1960 and over the following 40 years was a problem confined largely to the healthcare setting. In the late 1990s the first US reports of so-called community-associated MRSA (CA MRSA) infections appeared. CA-MRSA infections were defined as MRSA infections occurring in patients who had no identifiable predisposing risk factors, such as healthy children and young adults. CA-MRSA is associated with a novel genetic profile and phenotype; it is remarkably fit and capable of spreading within communities, it is virulent and is often susceptible to multiple narrow-spectrum antimicrobials other than beta-lactams. CA-MRSA infections involve predominantly skin and soft tissue; however, necrotizing pneumonia and necrotizing fasciitis have been described. At present, several reports suggest that CA-MRSA may be replacing the hospital-acquired MRSA strains (HA-MRSA), with potentially catastrophic consequences. Given the rapid spread and the high virulence of CA MRSA, global strategies are needed. Prompt, appropriate treatment, guided by the site and type of infection and risk factors for HA-MRSA or CA-MRSA, increases the chances of a successful outcome and is urgently needed. PMID- 19560670 TI - Skin and soft tissue infection: microbiology and epidemiology. AB - Skin and soft tissue infections (SSTIs) are common and range in severity from minor, self-limiting, superficial infections to life-threatening diseases requiring all the resources of modern medicine. The classification of SSTIs can be based on the anatomical site, clinical severity or microbial cause, but some classifications divide SSTIs into complicated and uncomplicated infections. Community-acquired SSTIs are most commonly caused by staphylococci or streptococci, but almost any organism is capable of causing inflammation within soft tissue. Recent epidemiological trends have shown an increase not only in healthcare-associated meticillin-resistant Staphylococcus aureus (MRSA), but also in MRSA acquired in the community. Many of the latter strains produce exotoxins and are epidemiologically distinct from healthcare-acquired strains. Factors that may affect the microbial cause include underlying disease such as diabetes or immune dysfunction; hospital attendance, injecting drug use, travel, animal contact and environmental contamination. PMID- 19560671 TI - Treatment options for skin and soft tissue infections: 'oldies but goldies'. AB - Although much is spoken and written about the requirement for novel antibiotics to treat serious infections, certain well-established agents are still valuable for the treatment of many infections, including those of the skin and soft tissue. These will be discussed in this paper. PMID- 19560672 TI - New antibiotics for the management of complicated skin and soft tissue infections: are they any better? AB - A large number of novel antibacterial agents have been or are being developed for the treatment of complicated skin and soft tissue infections (cSSTIs). Only linezolid, daptomycin and tigecycline are available for clinical use. The clinical decision-making process around antibiotic choice in the empirical setting or where cultures are available is dependent on a variety of factors. Important parameters that appear to determine the clinical effectiveness of an antibiotic for cSSTIs include the severity of the illness, patient co morbidities, whether the patient receives appropriate antimicrobial therapy at the onset of illness and if this should be a combination or single-agent approach to cover a broad range of likely causative organisms. The importance of these is discussed and the effectiveness of the new agents against current standard-of care comparators is measured using a number of outcomes. The data presented do not provide a clear and robust case for widespread empirical use of new agents, but suggest a more measured, patient-centred, risk-benefit approach to antibiotic choice. PMID- 19560673 TI - Treatment options for skin and soft tissue infections caused by meticillin resistant Staphylococcus aureus:oralvs.parenteral; home vs. hospital. AB - Although meticillin-resistant Staphylococcus aureus (MRSA) is recognized as a significant cause of nosocomial infections, it is also becoming an increasingly common cause of infections in the community. Overall, the most common infections caused by MRSA are those involving the skin and skin structures. These infections are difficult to treat and are associated with high morbidity and substantial cost. This article summarizes the current oral and parenteral therapeutic options, of which there are several, and the optimal site of care for the management of these infections. Defining the severity of the illness is central to improving the decision-making process about the route of administration and site of care. PMID- 19560674 TI - Review of question and answer session. AB - Prior to the oral presentations at the Skin and Soft Tissue Infections Workshop the audience was polled to assess the participants' knowledge of the epidemiology and aetiology of skin and soft tissue infections (SSTI) as well as to determine their approach to the treatment of complicated SSTIs (cSSTI). Twenty questions were asked before the presentations and audience members were allowed to answer anonymously using an electronic audience response system. The questions fell into four general categories: aetiology of cSSTI, terminology, epidemiology and treatment. This paper discusses the answers to the questions. PMID- 19560675 TI - Antibiotics, skin and soft tissue infection and meticillin-resistant Staphylococcus aureus: cause and effect. AB - Staphylococcus aureus is not a new cause of skin and soft tissue infection, but the significance of the Panton-Valentine leukocidin toxin and the spread of several clones carrying different staphylococcal cassette chromosome (SCC) mecA gene cassette types have given it a new lease of life. This is a clinical area with several epidemic strains causing major problems around the world, most notably in the USA. While most attention focuses on treatment, prevention should be the goal. Traditional infection control measures, such as good hand hygiene and barrier precautions, are usually emphasised. Most importantly, we should not forget the underlying cause of meticillin-resistant Staphylococcus aureus (MRSA), namely antibiotic use. In both community and hospital, exposure to beta-lactams, in particular cephalosporins, and also sometimes quinolones and macrolides, is likely to promote the transmission, colonisation and increased virulence of MRSA. Future antibiotic policies should consider this, particularly in an era of widespread MRSA screening. PMID- 19560676 TI - Aphasia among young patients with ischemic stroke on long-term follow-up. AB - BACKGROUND: The aim of this study was to evaluate characteristics and long-term outcome of young patients with ischemic stroke (15-49 years at stroke onset) and aphasia. METHODS: Aphasia was defined as less than 10 points in the speech subscale of the Scandinavian Stroke Scale on long-term follow-up. Risk factors, origin, complications, social factors, and the Nottingham Health Profile subscores were compared between aphasic and nonaphasic young patients with ischemic stroke. On long-term follow-up, patients with aphasia were invited for further assessment of severity and subtype of aphasia. RESULTS: The study comprised 195 patients still alive after a mean follow-up of 6 years. Twenty (10.3%) patients had aphasia. Aphasia was associated with cardiac embolism (P = .007), myocardial infarction (P = .005), epilepsy (P < .001), loss of employment (P = .021), and social isolation (P = .054). On follow-up, 13 patients with aphasia underwent further assessment. These patients had all improved into milder aphasia subtypes. CONCLUSION: Our study suggests that relatively few young patients with ischemic stroke have clinically significant aphasia on long-term follow-up. However, there are clinically significant differences between patients with and without aphasia that should be the focus of future research. PMID- 19560677 TI - Predictors of stroke and its significance in the outcome of tuberculous meningitis. AB - BACKGROUND AND AIM: We sought to study the frequency and predictors of stroke in tuberculous meningitis (TBM) and its prognostic significance. DESIGN: This was an observational study in a tertiary care teaching hospital. METHODS: In all, 122 patients with TBM aged 4 to 82 years diagnosed on the basis of clinical, cerebrospinal fluid, and magnetic resonance imaging criteria were prospectively evaluated. Severity of meningitis was graded into stage I to III. Magnetic resonance imaging was done at admission and 3 months after treatment. Outcome was defined at 3 and 6 months as complete, partial, or poor. Predictors of stroke and its significance in long- and short-term outcome were evaluated. FINDINGS: A total of 55 patients had stroke; 42 at admission and 13 developed within 3 months of 4 drug antitubercular treatment. Strokes were ischemic in 54 (hemorrhagic transformation in 7) and hemorrhagic in one. Basal ganglia infarctions were present in 30, thalamic in 9, brainstem in 10, cortical in 27, and cerebellar in 4 patients. Stoke was multiple in 29 patients. In all, 38 patients had infarctions in anterior circulation, 7 in posterior, and 10 in both. Stroke was significantly related to stage of meningitis, hydrocephalus, exudate, and hypertension. No difference was found in clinical or laboratory parameters in early and late strokes. At 6 months, 28 patients died. At 3 months there were 21 patients lost to follow up and at 6 months there were 30 patients lost to followup. Outcome is based on the rest of the patients, ie. 101 patients at 3 months and 92 patients at 6 months. CONCLUSION: Stroke occurs in 45% of patients with TBM both in early and later stage, mostly in basal ganglia region, and predicts poor outcome at 3 months. PMID- 19560678 TI - Stroke and newspapers: inattention or neglect? AB - BACKGROUND: Services for stroke are poorly developed in Europe, and research into stroke is underfunded compared with heart disease or cancer. This may arise from the low profile of stroke within the public domain. Our aim was to assess the coverage of stroke compared with heart disease in a popular form of mass media, newspapers in the United Kingdom and Ireland. METHODS: We searched the online databases of 4 major newspapers in the United Kingdom and Ireland using the terms "stroke" and "heart" during a 1-year period from May 1, 2006, to May 1, 2007. The resulting articles were then classified by two clinicians and those relating to the clinical conditions of stroke and heart disease were selected. RESULTS: The searches were refined by hand and those articles relating to stroke were: 105 in the Irish Times, 223 in the Daily Telegraph, 22 in The Observer, and 149 in The Guardian. The number of articles relating to heart disease was: 443 in the Irish Times, 653 in the Daily Telegraph, 81 in The Observer, and 688 in The Guardian. CONCLUSIONS: Newspaper coverage of stroke is underreported when compared with heart disease, despite the clinical and financial burden of this condition. Professional and advocacy organizations for those with stroke need to develop strategies to increase the profile of stroke in the public domain. PMID- 19560679 TI - Somatosensory evoked potentials as a predictor for functional recovery of the upper limb in patients with stroke. AB - BACKGROUND: Predicting motor recovery in the arm of patients with stroke is generally based on clinical examination. However, neurophysiologic measures may also have a predictive value. OBJECTIVE: We sought to assess the value of somatosensory evoked potentials (SSEPs) in predicting motor recovery of the upper limb and to determine whether any of the SSEPs components can predict the severity of the deficit so that it can document the size of the stroke (lacunar or large-vessel stroke). METHOD: In all, 22 patients who had had a first-ever stroke and presented with obvious motor deficit of the arm were examined in terms of 3 clinical variables (motor performance, muscle tone, and overall disability) and for SSEPs. Clinical (Medical Research Council [MRC] scale and Barthel index scores) and neurophysiologic examinations were done at entry to the study (first week poststroke) and 3 months after stroke. RESULTS: Significantly low mean MRC scale score was found at first week versus after 3 months of stroke and in patients with large-vessel as compared with lacunar stroke both at first week and after 3 months. The mean Barthel index score was significantly higher after 3 months than at first week, whereas it was significantly lower in large-vessel as compared with lacunar stroke both at first week and after 3 months of stroke. Significantly prolonged N(20) latency, low peak-to-peak amplitude (PPA), and low amplitude ratio were found in patients with stroke as compared with control subjects. None of the neurophysiologic parameters were different in the patients with stroke between first week and third month. The MRC score and PPA were correlated well with the outcome MRC and Barthel index scores after 3 months. N(20) latency correlates with the outcome MRC score but not with the outcome Barthel index score. Interestingly, the N(20) latency was significantly different in lacunar from large-vessel stroke. CONCLUSION: The muscle power (MRC score) is the main outcome predictor in patients with stroke. PPA is the main SSEPs component with high prognostic value in stroke. The SSEPs N(20) latency can predict (even roughly) the size of cerebral infarction (whether lacunar or large vessel stroke). PMID- 19560680 TI - Systematic review and stratified meta-analysis of the efficacy of interleukin-1 receptor antagonist in animal models of stroke. AB - BACKGROUND: Interleukin (IL)-1 receptor antagonist (RA) is an anti-inflammatory protein used to treat arthritis that has also been identified as a candidate stroke drug. METHODS: We conducted a systematic review and meta-analysis of reports of the efficacy of IL-1 RA in animal models of focal cerebral ischemia. RESULTS: We identified 16 published sources and one unpublished source of data. IL-1 RA reduced infarct volume by 38.2% (95% confidence interval 31.2%-45.1%). Efficacy was higher with higher doses, earlier treatment, and central administration of drug. No studies used animals with hypertension or diabetes or tested efficacy beyond 3 hours. CONCLUSIONS: The animal data supporting IL-1 RA as a candidate drug for stroke are limited, and further experiments are required before proceeding to clinical trial. PMID- 19560681 TI - High rate of inappropriate carotid endarterectomy in an urban medical center. AB - BACKGROUND: Some studies have reported that the appropriateness of carotid endarterectomy (CEA) has dramatically improved in the last two decades. However, these studies did not include the most recent study results for asymptomatic stenosis. METHODS: We sought to determine the appropriateness of CEA at a large, urban community hospital. A retrospective chart review of all CEA procedures during a 2-year period was conducted. CEA appropriateness was defined according to factors including age, degree of stenosis, symptomatic or asymptomatic status, and presence of high surgical risk comorbidities. RESULTS: During a 2-year period, 51 CEA procedures were performed. For patient characteristics, 73% were asymptomatic, 77% were African American, and 65% had high surgical risk comorbidities. Perioperative stroke or myocardial infarction occurred in 14.3% of symptomatic patients and 18.9% of asymptomatic patients. The rate of inappropriate CEA was 57%, and 14% of cases were of uncertain value. Patients with an inappropriate or uncertain CEA had a 19.4% rate of in-hospital stroke or myocardial infarction. CONCLUSIONS: In an urban hospital, the majority of CEA procedures were either inappropriate or of uncertain value. The periprocedure complication rate was high in these patients. Patient selection in urban hospitals is not adhering to clinical trial criteria. PMID- 19560682 TI - Experimental model of brainstem stroke in rabbits via endovascular occlusion of the basilar artery. AB - BACKGROUND: Basilar artery thrombosis remains a significant clinical problem, and no reproducible animal model has been established to study the stroke within the vertebrobasilar distribution. We report a study designed to pilot test a novel model of brainstem stroke in rabbits, created by selective endovascular occlusion of the basilar artery. METHODS: Basilar artery occlusion was induced in 8 New Zealand white rabbits by injection of the autologous clot through the microcatheter positioned within the distal vertebral artery. Animals were divided into subgroups (I and II) based on the length of produced ischemia (3 and 6 hours, respectively). Magnetic resonance (MR) imaging of the brain and MR angiography of the intracranial vessels were performed before the procedure, and at 3 hours after induced ischemia for groups I and II, with continued imaging up to 6 hours for group II, with diffusion-weighted images acquired approximately every 30 minutes. Animals were killed at the end of the 3-hour (group I) or 6 hour (group II) ischemia time. RESULTS: Brainstem stroke was successfully induced in all animals, with pathological changes documented in all cases. The earliest changes of ischemia on MR diffusion-weighted images were identified at only 4.5 hours of basilar artery occlusion. CONCLUSION: These results suggest that a reproducible model of brainstem stroke can be induced in rabbits using selective endovascular occlusion of the basilar artery. The availability of such a model, integrated with state-of-the-art imaging techniques, holds promise for preclinical investigations of emergent therapeutic approaches in stroke. PMID- 19560683 TI - The "GP" mechanical thrombectomy device. AB - We present data concerning the extraction of clots using the newly invented "GP" mechanical thrombectomy device (MTD). Artificial and porcine clots of various lengths were used in plastic tube models of an artery. We investigate the pressures and times taken for clot extraction together with the volumes of fluid extracted. We also investigate the impact of using a funnel structure mounted on the end of the device, on clot removal times and fluid removed. Finally, we present results involving clot extraction from the posterior popliteal artery of a cadaver. Our data indicated that: The embedded GP MTD is the most effective device regarding artificial and porcine blood clot removal. This result is consistent with previous published data on this device. The GP MTD was effective in removing clots positioned in the posterior popliteal artery of a cadaver. The embedded GP device removes less fluid compared with the end-mounted GP device. This confirms previous studies. There appears to be a relationship between funnel angle and pressure. Lower extraction pressures are required for larger funnel angles mounted on the GP device. Shorter times of clot extraction are required for larger funnel angles. PMID- 19560684 TI - Is insulin resistance related to recurrence of stroke or incident of ischemic heart disease in patients with stroke? A preliminary report. AB - Insulin resistance promotes atherosclerosis and has been recognized as an important risk factor for both stroke and ischemic heart disease (IHD). However, the precise relationship between insulin resistance and ischemic stroke recurrence or development of IHD is uncertain. The aim of this study was to investigate the relationship of insulin resistance to ischemic stroke recurrence or IHD in patients with a history of stroke. A total of 32 patients poststroke with hypertriglyceridemia but without a history of diabetes mellitus were asked whether they had experienced stroke recurrence or IHD after discharge. We found that 4 of 32 patients experienced recurrence. Three of these 4 patients had insulin resistance. We classified the subjects into two groups: one group with stroke recurrence and the other group without. We then compared the insulin profiles between these two groups. Although there was no significant relationship between recurrence and insulin resistance, fasting glucose level was higher in patients with recurrence than without. Insulin-resistant patients with recurrence had significantly lower levels of plasma insulin at 120 minutes in the oral glucose tolerance test compared with those without recurrence. This study suggests that insulin resistance in combination with decreased insulin secretion may be associated with ischemic stroke recurrence or IHD. PMID- 19560685 TI - Spinal cord infarction in congenital afibrinogenemia: a case report and review of the literature. AB - A 22-year-old woman with congenital afibrinogenemia presented with acute-onset rapidly progressive quadriparesis as a result of spinal cord infarction caused by vertebral artery dissection. Magnetic resonance imaging scans showed microhemorrhages in addition to edematous swelling suggesting acute ischemia throughout cervical and upper thoracic portions of the spinal cord. Fat-saturated T1-weighted magnetic resonance examination and digital subtraction angiography studies demonstrated cervical vertebral artery dissection on the right. This case provides an example of how a primary bleeding disorder could result in a severe ischemic complication caused by an occlusive vessel wall hematoma. Along with other reports, diagnostic and therapeutic aspects of this paradoxical situation were discussed in the particular setting of acute spinal cord ischemia. PMID- 19560686 TI - Young strokes in Sri Lanka: an unsolved problem. AB - Stroke in young adults is more common in India and Sri Lanka and the reasons for this are not well understood. The current study was conducted to elucidate the risk factors and radiologic features in young people (age < 45 years) with ischemic stroke. Sociodemographic data, stroke risk factor information, and laboratory investigations were recorded in 41 cases with first-ever ischemic stroke. Most common risk factors for stroke in the 15- to 45-year-old age group were: hypertension, 8 (21%); family history of stroke, 7 (18%); transient ischemic attack, 6 (16%); hyperlipidemia, 3 (8.0%); and diabetes, two (5%). Age group younger than 15 years included 3 girls and one had a mass attached to the posterior mitral valve leaflet. Our observations underscore the importance of the presence of hypertension, family history of stroke, and transient ischemic attack in young adults and thus to adopt preventative strategies. PMID- 19560688 TI - Bilateral subdural hematomas in an adult with osteogenesis imperfecta. AB - Osteogenesis imperfecta (OI) is a rare inherited disorder characterized by disruption of normal collagen formation, brittle bones, and low bone density, predisposing to multiple bone fractures and wide-spectrum symptomatology. Extradural and subdural hematomas and spinal cord injuries have been reported but mainly in children with OI. We cared for a 60-year old woman with OI and bilateral nontraumatic subdural hematomas possibly caused by spontaneous intracranial hypotension. PMID- 19560687 TI - Brainstem ischemia, steno-occlusive pathology of the vertebral arteries, and alterations in the circadian blood pressure pattern: a case report. AB - We report the case of a patient with vertebrobasilar steno-occlusive pathology associated with ischemic brainstem symptomatic lesions related to abnormal decrease in blood pressure (BP). A 63-year-old man presented vertigo and persistent gait impairment as a result of a posterior cerebellar infarction and, during hospitalization, experienced a new episode of cerebral ischemia in the territory of the anterior cerebellar artery. Cerebral angiography showed occlusion of the left vertebral artery and two severe stenoses of the right vertebral artery, and 24-hour dynamic BP monitoring evidenced severe postprandial hypotension. Medical treatment was not effective so he underwent an endovascular procedure. This report documents the importance of BP monitoring for a better understanding of the pathophysiology of posterior ischemic strokes and, above all, for a more reliable prognosis and therapy. PMID- 19560689 TI - Acute massive cerebral infarctions treated with hemodialysis. AB - We report here a 19-year-old woman with Down syndrome and end-stage renal disease who presented with left-sided weakness and fever. She had a massive pericardial effusion of unclear origin that required daily hemodialysis (HD) and cardiac intervention. She developed an acute right middle cerebral artery infarction with severe edema; her cerebral edema significantly improved with daily HD. Later in her hospitalization, she developed seizures and new onset of multiple acute embolic infarctions in left middle cerebral artery, left anterior cerebral artery (ACA), and right posterior cerebral artery (PCA) distributions with midline shift. However, we again noticed a dramatic decrease in cerebral edema with frequent HD. Although there is controversy about the use of dialysis in patients with stroke, our case suggests that daily HD may provide an alternate strategy for treating massive cerebral infarction. More studies are needed in these patients. PMID- 19560690 TI - Carotid artery ultrasound and echocardiography testing to lower the prevalence of Alzheimer's disease. AB - The use of two clinic office techniques, carotid artery ultrasound and echocardiography (CAUSE), to detect deficient brain blood flow delivery in the healthy, cognitively normal, older individual is proposed. Evidence indicates that persistent heart-to-brain blood flow deficit involving low cardiac output or low ejection fraction and carotid artery narrowing can promote cognitive impairment and that such impairment may lead to Alzheimer's disease (AD) or vascular dementia (VaD). A number of independent epidemiologic studies reported cardiovascular and cerebrovascular disease to be risk factors to AD and VaD. The clinical rationale for CAUSE is to detect and prevent progression of cognitive dysfunction in elderly persons and is based on the general understanding that mild cognitive impairment is a preclinical threshold to AD or VaD with high conversion rates to either dementia. The use of CAUSE is anticipated to prevent or attenuate, by appropriate clinical management, mild cognitive impairment arising from persistent brain hypoperfusion, a condition implicated in the promotion of cognitive impairment in the elderly and a common preclinical feature seen in AD and VaD. As detection of lowered cerebral perfusion from cardiac and carotid artery pathology is achieved using these cost-effective, noninvasive, and relatively accurate ultrasound procedures, a significant reduction in the number of new AD and VaD cases would be anticipated after appropriate patient treatment. In this context, a brief summary is presented outlining recent medical and surgical treatments that may improve cerebral blood flow insufficiency. The merit of CAUSE in screening and treating mentally healthy elderly persons who are identified as being at risk of cognitive decline is that it could blunt the soaring socioeconomic impact that will hammer the health care system of many nations by the mounting dementia prevalence rate expected in the next 25 years. A 5- year delay in the onset of AD could reduce the prevalence of AD by 50%. We are making preparations to test the merit of CAUSE in a clinical study of a cognitively healthy elderly population. PMID- 19560691 TI - Generic drug immunosuppression in thoracic transplantation: an ISHLT educational advisory. PMID- 19560692 TI - Quality of life in adult survivors greater than 10 years after pediatric heart transplantation. AB - BACKGROUND: This study assessed quality of life (QOL) in adult survivors of pediatric heart transplantation who survived > or = 10 years after transplantation. METHODS: Prospective data were collected from heart transplant recipients who were aged > or = 18 years and had survived > or = 10 years after transplantation (transplantation between July 3, 1986, and April 4, 1997). QOL data were collected from patients using the Medical Outcomes Study 36-Item Short Form (SF-36) Health Survey. Clinical data were collected from medical records. Statistical analyses included frequencies and measures of central tendency. RESULTS: Twenty-three patients (65% men, 91% white) completed the study. At the study initiation, they were a mean age of 9.0 +/- 7.1 years at transplantation, and were a mean age of 25.2 +/- 5.5 years (range, 18-34 years) and a mean of 16.2 +/- 3.0 years (range, 11-22 years) post-transplantation. Most were in school or working. Mean patient QOL scores from the SF-36v2 survey were 50.56 +/- 0.5 (range, 27.3-68.9) for physical health and 49.88 +/- 11.72 (range, 23.56-62.84) for mental health, similar to the general United States population. Late complications were frequent, including transplant coronary artery disease, 3; repeat heart transplantation, 2; post-transplantation lymphoproliferative disorder, 6; kidney transplantation, 5; acute late rejection, 5; and arrhythmias, 4. CONCLUSION: This report of QOL in adult survivors of pediatric heart transplantation shows patient perception of physical and mental health is similar to the general population despite serious late complications. A multicenter study is planned to further evaluate QOL in this unique cohort. PMID- 19560693 TI - Heart transplantation in human immunodeficiency virus-positive patients. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection is widely considered a contraindication for cardiac transplantation. However, with the newer anti retroviral drugs, the estimated 10-year survival after seroconversion is exceeds 90%. This case series describes the intermediate range outcome of HIV-positive cardiac transplant recipients. METHODS: A retrospective analysis of 1679 cardiac transplant patients was undertaken to identify HIV-positive recipients. RESULTS: Seven patients were identified. Five (4 men) were diagnosed with HIV before transplantation and 2 patients seroconverted after transplantation. Dilated cardiomyopathy was the indication for transplant in all patients. The 5 HIV recipients were aged 42 +/- 8 years, and time after HIV seroconversion averaged 9.5 years. All underwent cardiac transplantation as high-risk candidates. The CD4 count was 554 +/- 169 cells/microl, and viral load was undetectable in all patients at the time of transplantation. Two patients seroconverted to HIV positive status at 1 and 7 years after transplant. No AIDS-defining illness was observed in any patient before or after transplant. Six patients received highly active anti-retroviral therapy. Viral load remained low in the presence of immunosuppression. All patients are alive with a follow-up from transplant of 57 +/- 78.9 months. CONCLUSION: Excellent intermediate term outcome is noted in carefully selected HIV-positive patients. No significant AIDS-related infections or complications occurred. PMID- 19560694 TI - Cytomegalovirus prevention in high-risk lung transplant recipients: comparison of 3- vs 12-month valganciclovir therapy. AB - BACKGROUND: Cytomegalovirus (CMV) infections are common after lung transplantation (LuTx) and have an influence on acute rejection rates and chronic organ dysfunction. The objective of this study was to determine the incidence of CMV infections by comparing a prolonged valganciclovir prophylaxis with a standard regimen in high-risk LuTx recipients. METHODS: A retrospective, single center study was performed comparing two different CMV prophylactic regimens in high-risk LuTx recipients (D(+)/R(-)). The study population received either 3 months (Group A, 15 patients) or 12 months (Group B, 17 patients) of oral valganciclovir 900 mg/day in combination with CMV hyperimmune globulin in four doses (Days 1, 7, 14 and 21 post-transplant). RESULTS: CMV viremia was noted in 11 of 15 patients in Group A (75%) and 5 of 17 in Group B (33%) (p < 0.05) at 6 months after valganciclovir cessation. The incidence of symptomatic CMV disease/syndrome was 6 of 15 (44%) in Group A and 2 of 17 in Group B (13%) (p < 0.05). Histologically proven acute rejection episodes of ISHLT Grade > or =A2 were found in 4 patients in Group A and in 1 patient in Group B within the first year (p = 0.14). CONCLUSIONS: A 12-month CMV prophylaxis with oral valganciclovir is effective in significantly reducing CMV viremia and CMV disease/syndrome in high-risk lung transplant recipients. In addition, a reduction in acute and recurrent rejection episodes was observed, possibly due to less CMV viremia and subsequent immunomodulatory effects. PMID- 19560695 TI - Acute hemodynamic effects of intravenous sildenafil citrate in congestive heart failure: comparison of phosphodiesterase type-3 and -5 inhibition. AB - BACKGROUND: The reversibility of elevated pulmonary vascular resistance in heart failure bears an important relation to outcome after cardiac transplantation. The phosphodiesterase 3 (PDE3) and PDE5 inhibitors both increase levels of cyclic nucleotides in the vascular smooth muscle, causing vasodilatation. PDE3 inhibitors also have direct inotropic effects. We contrasted the acute hemodynamic responses to intravenous PDE3 and PDE5 inhibitors in patients with congestive cardiac failure to assess their relative suitability for reversibility testing in this setting. METHODS: Thirty patients undergoing assessment for cardiac transplantation underwent right heart catheterization. Patients were randomized to receive an intravenous bolus of milrinone (0.05 mg/kg) or sildenafil citrate at a high (0.43 mg/kg) or low dose (0.05 mg/kg). RESULTS: Differences between low- and high-dose sildenafil were not significant. Both agents caused similar reductions in systemic and pulmonary vascular resistance. Milrinone caused significantly greater reductions in pulmonary artery wedge and mean pulmonary artery pressure, and increases in heart rate. In all study groups, greater increases in cardiac index (>25%) were seen in patients with a higher pulmonary artery wedge pressure at baseline (29 +/- 1 vs 20 +/- 2 mm Hg; p < 0.001). CONCLUSIONS: In end-stage congestive cardiac failure, intravenous milrinone and sildenafil both cause similar reductions in systemic and pulmonary vascular resistance; however, milrinone has more cardiac selective effects on left ventricular filling and heart rate. Both agents appear to have a suitable hemodynamic profile for testing of reversibility of secondary pulmonary hypertension in congestive cardiac failure. Larger studies are needed to confirm these results. PMID- 19560696 TI - Clinical experience with a new removable tracheobronchial stent in the management of airway complications after lung transplantation. AB - BACKGROUND: Airway complications are among the most challenging problems after lung transplantation. This article describes the use of a new tracheobronchial stent that can be placed and removed easily by flexible bronchoscopy. METHODS: A retrospective review was done of 24 consecutive patients requiring tracheobronchial stent placement after lung transplantation. A new self-expanding hybrid nitinol stent was used, and changes in airway diameter and spirometry were assessed. Stent related complications were recorded. RESULTS: Between February 2007 and April 2008, 24 patients underwent stent placement, and 49 stents were placed for 36 anastomoses at risk. Indications included bronchial stenosis in 12, bronchomalacia in 12, bronchial stenosis plus bronchomalacia in 20, and partial bronchial dehiscence in 5. Adjunctive procedures included electrocautery in 1, balloon dilatation in 7, and electrocautery plus balloon dilatation in 4. The average degree of stenosis decreased from 80% to 20%. After stent placement, the average increase was 0.28 liters in forced vital capacity and 0.44 liters in forced expiratory volume in 1 second. Complications included granulation tissue formation in 10 stents, migration in 9, thick mucus formation in 2, and fracture in 3. CONCLUSION: Airway complications in lung transplant patients were effectively palliated. Our complication rate with this new stent is comparable with other airway stents. This stent has the advantage of easy removability during flexible bronchoscopy if complications from the stent outweigh the benefits of palliation. PMID- 19560697 TI - Microvolt T-wave alternans, peak oxygen consumption, and outcome in patients with severely impaired left ventricular systolic function. AB - BACKGROUND: Abnormal microvolt T-wave alternans (MTWA) and low peak oxygen consumption (VO2) both predict poor outcome in heart failure. However, their independent predictive properties have not been assessed in large-scale cohorts. METHODS: This was an observational prospective cohort study of 303 consecutive patients referred for metabolic stress testing. All had an ejection fraction < or = 40% and were considered candidates for transplantation. The exercise laboratory did not collect MTWA data from patients with implanted pacemakers or defibrillators. The primary end point was a composite of all-cause death or United Network for Organ Sharing status 1 transplantation. RESULTS: During a 2.8 year period, there were 34 deaths and 17 transplantations. Patients with abnormal MTWA had a higher event rate of 23% (31 of 136) vs 12% (20 of 167), with an unadjusted hazard ratio (HR) of 1.90 (95% confidence interval [CI], 1.90-3.33; p = 0.03). The association remained significant after adjustment for 3 clinical variables (HR, 1.89; 95% CI, 1.05-3.39; p = 0.03). After adding peak VO2 to the model, the association was no longer significant (adjusted HR, 1.18; 95% CI, 0.64 2.17, p = 0.60). After accounting for peak VO2 and 28 other confounders in a matched propensity analysis, MTWA was not predictive (propensity-matched HR, 0.79; 95% CI, 0.37-1.66; p = 0.53). CONCLUSIONS: These results confirm the association of abnormal MTWA with poor outcome amongst patients with impaired left ventricular systolic function. However, this association is markedly attenuated after accounting for peak VO2. PMID- 19560698 TI - Tacrolimus and azathioprine versus cyclosporine and mycophenolate mofetil after lung transplantation: a retrospective cohort study. AB - BACKGROUND: The efficacy and safety of different combinations of immunosuppressive regimens after lung transplantation are unknown. METHODS: We examined 120 consecutive transplant recipients between July 2001 and July 2005, of whom 37 received cyclosporine and mycophenolate mofetil (Cyc/MMF) and 83 received tacrolimus and azathioprine (Tac/Aza) as the initial immunosuppressive regimen along with an interleukin-2 antagonist induction therapy. The primary outcome was the rate of histologically confirmed acute rejection. RESULTS: The rate of acute rejection did not vary by treatment regimen (0.42 vs 0.34 episodes per 100 person-days in Cyc/MMF and Tac/Aza groups, respectively, p = 0.22). The mean cumulative lymphocytic bronchiolitis score was greater in the Cyc/MMF group (1.8 +/- 1.9) compared with the Tac/Aza group (1.2 +/- 2.0; p = 0.03). Pulmonary function at 1 year was better in the Tac/Aza group, even when adjusted for recipient age, gender, and transplant procedure. Survival and the rate of bronchiolitis obliterans syndrome did not vary by group. CONCLUSIONS: Outcomes after lung transplantation did not meaningfully vary between those assigned to Cyc/MMF compared with Tac/Aza combined with IL-2 inhibitor induction therapy. PMID- 19560699 TI - The change in B-type natriuretic peptide levels over time predicts significant rejection in cardiac transplant recipients. AB - BACKGROUND: B-type natriuretic peptide (BNP) correlates with cardiac filling pressures and outcomes in patients with heart failure. In heart transplant recipients, we hypothesize that a within-individual change in BNP over time would be more helpful than absolute BNP in detecting International Society of Heart and Lung Transplantation (ISHLT) grade 2R or greater rejection. METHODS: N-terminal pro-BNP (NT-proBNP) levels were measured in 146 consecutive transplant recipients undergoing routine endomyocardial biopsies. In the cross-sectional analysis, multiple observations per individual were accounted for using generalized estimation equations. RESULTS: A cross-sectional analysis demonstrated a weak association between NT-proBNP levels and rejection, with an odds ratio (OR) of 1.01 for every 100-pg/mL increase in NT-proBNP (p = 0.02). However, with a doubling of an individual's NT-proBNP level, the OR for significant rejection was 2.9 (95% confidence interval [CI] 1.2-7.0), the OR with a 5-fold increase was 9.1 (95% CI, 2.7-31.5), and the OR with a 10-fold increase was 27.7 (95% CI, 5.9 129). A 10-fold increase in NT-proBNP offered a negative predictive value of 95% for the diagnosis of rejection. The relationship between within-individual increases in NT-proBNP and rejection persisted after adjusting for a fall in ejection fraction and a rise pulmonary capillary wedge pressure, and was a stronger predictor than changes in these parameters. CONCLUSIONS: There is a strong, graded relationship between the within-individual increase in NT-proBNP and the odds of significant rejection independent of hemodynamic parameters. These results suggest that the change in NT-proBNP rather than absolute BNP levels may offer a non-invasive approach to detect rejection. PMID- 19560700 TI - Increased number of circulating progenitor cells after implantation of ventricular assist devices. AB - BACKGROUND: Bone marrow-derived circulating progenitor cells possess tissue repair potential, improving perfusion, left ventricular remodeling, and contractility in experimental models. We quantified and investigated the kinetics of 4 circulating progenitor cell sub-populations on the basis of CD34, CD133, and vascular endothelial growth factor receptor-2 (VEGFR-2) antigen expression. METHODS: CD34+, CD34+/CD133+/VEGFR-2-, CD34+/CD133+/VEGFR-2+, and CD34+/CD133 /VEGFR-2+ cells were counted in 10 male patients with end-stage congestive heart failure. Five underwent left ventricular/biventricular assist device (LVAD/BiVAD) implantation (VAD group), and 5 were ineligible for VAD implantation (no-VAD group). Peripheral blood was collected at 3 time points for each patient: before, 15, and 60 days after VAD placement in the VAD group and at the same time points in the no-VAD group. Purified CD34+ cells were stained with anti-CD34, anti CD133, and anti-VEGFR-2 monoclonal antibodies and analyzed by flow cytometry. Serum levels of granulocyte-colony stimulating factor (G-CSF), interleukin-8, vascular endothelial growth factor-alpha (VEGF-alpha), and B-type natriuretic peptide (BNP) were also measured. RESULTS: In the VAD group the number of CD34+ cells/ml of blood tended to increase, from 159.6 +/- 137.0 at baseline to 428.9 +/- 224.3 at 15 days, and decreased to 343.8 +/- 165.7 at 60 days (p = 0.05 vs no VAD group). In the other 3 cell populations, no significant differences occurred over time or between groups. A significant interaction between BNP levels and VAD status was observed (p = 0.005): BNP levels decreased over time in VAD patients vs no-VAD patients. G-CSF levels tended to decrease over time in both groups, but without a significant difference (p = 0.3). Serum levels of interleukin-8 and VEGF-alpha over time or between VAD and no-VAD patients were not significantly different. CONCLUSIONS: After VAD implantation, a transient increase occurs in the number of circulating CD34+ cells, in parallel to a reduction in BNP levels. Release of these cells from the bone marrow may contribute to the improvement of tissue perfusion and cardiac recovery occasionally seen after VAD placement. PMID- 19560701 TI - Methylprednisolone and tacrolimus prevent hypothermia-induced endothelial dysfunction. AB - BACKGROUND: Hypothermia is used to preserve organs for transplantation and is the oldest method to protect organs during complex pediatric cardiac surgery. Loss of tissue function and tissue edema are common complications in children undergoing corrective cardiac surgery and heart transplantation. The present study was designed to examine the effects of methylprednisolone and tacrolimus on endothelial cell function and morphology after deep hypothermia and rewarming. METHODS: Human umbilical vein endothelial cells were pre-treated with methylprednisolone or tacrolimus, or both, incubated within a specially designed bioreactor or in monolayers, and then exposed to a dynamic cooling and rewarming protocol. Immunocytochemistry, time-lapse video microscopy, cell permeability and adherence assays, and Western blot analysis were performed. RESULTS: Confluent endothelial cells exposed to hypothermia displayed elongated cell shapes with intercellular gap formation, increased endothelial cell-layer permeability, and loss in adherence. Upon rewarming, however, endothelial cell integrity was restored. Opening and closing of intercellular gaps was dependent on extracellular signal-regulated kinase 1 and 2 (ERK 1/2) activation and connexin 43 expression. The combined treatment with methylprednisolone and tacrolimus inhibited these hypothermia-induced changes. CONCLUSIONS: These results suggest that methylprednisolone and tacrolimus inhibit hypothermia-induced endothelial gap formation by phosphorylated ERK 1/2 inhibition and connexin 43 stabilization. Application of combined drugs that affect multiple targets may therefore be considered as a possible new therapeutic strategy to prevent endothelial dysfunction after hypothermia and rewarming. PMID- 19560702 TI - Alteration of neuropeptides in the lung tissue correlates brain death-induced neurogenic edema. AB - BACKGROUND: Increased intracranial pressure induces neurogenic pulmonary edema (NPE), potentially explaining why only lungs from less than 20% of brain dead organ donors can be used for transplantation. This study investigated the underlying mechanisms of NPE, focusing on neuropeptides, which potently induce vasoconstriction, vasodilatation, and neurogenic inflammation. METHODS: Brain death was induced in 10 pigs by increasing the intracranial pressure. Eight additional pigs served as controls. Neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), and substance P were analyzed in plasma, bronchoalveolar lavage (BAL) fluid, and homogenized lung tissue 6 hours after brain death. Pulmonary oxygen exchange was estimated using partial pressure of arterial oxygen (Pao2)/fraction of inspired oxygen (Fio2), and pulmonary edema by wet/dry weight ratio. RESULTS: Brain death induced a decrease in Pao(2)/Fio2 (p < 0.001) and increased the wet/dry weight of both apical (p = 0.01) and basal lobes (p = 0.03). NPY and CGRP concentrations were higher in the BAL fluid of brain-dead animals compared with controls (p = 0.02 and p = 0.02) and were positively correlated with the wet/dry weight ratio. NPY content in lung tissue was lower in brain-dead animals compared with controls (p = 0.04) and was negatively correlated with the wet/dry weight ratio. There were no differences in substance P concentrations between the groups. CONCLUSION: NPY was released from the lung tissue of brain-dead pigs, and its concentration was related to the extent of pulmonary edema. NPY may be one of several crucial mediators of neurogenic pulmonary edema, raising the possibility of treatment with NPY-antagonists to increase the number of available lung donors. PMID- 19560703 TI - Incidence of ventricular arrhythmias in patients on long-term support with a continuous-flow assist device (HeartMate II). AB - The incidence of ventricular tachycardia (VT) or ventricular fibrillation (VF) in patients supported with a continuous-flow left ventricular assist device (LVAD) has not been investigated in detail. In 23 consecutive recipients of a HeartMate II, we analyzed the incidence of VT/VF during a total of 266 months of follow-up. Sustained VT or VF occurred in 52% of the patients, with the majority of arrhythmias occurring in the first 4 weeks after LVAD implantation. VT/VF requiring implantable cardioverter-defibrillator (ICD) shock or external defibrillation occurred in 8 patients and significant hemodynamic instability ensued in 3 patients. There were no clear predictors of VT/VF, and it is argued that prophylactic ICD implantation should be considered in patients supported with a continuous-flow LVAD. PMID- 19560704 TI - Disseminated infection with Bartonella henselae in a lung transplant recipient. AB - We present the case of a lung transplant recipient with disseminated infection with Bartonella henselae. In non-immunosuppressed humans, the organism typically causes a local infection that manifests itself as regional lymphadenopathy. The role of the host immune response to B henselae is critical in preventing progression to systemic disease. Only rare cases of bartonellosis in transplant recipients have been reported. We discuss aspects and difficulties of diagnosis and treatment of bartonellosis in a lung transplant recipient who suffered from a severe multisystem involvement of this disease. In our case, the initial response to therapy was unsatisfying and necessitated an extended anti-infective combination therapy, which eventually was successful. PMID- 19560705 TI - Ventricular assist device application with the intermediate use of a membrane oxygenator as a bridge to pediatric heart transplantation. AB - Extracorporeal membrane oxygenation and ventricular assist devices are currently used for the treatment of severe heart failure as a bridge to transplantation. The use of ventricular assist devices is limited by respiratory failure. We report a patient with severe heart failure and respiratory failure who was successfully bridged to transplantation, initially with extracorporeal membrane oxygenation and afterwards with an EXCOR biventricular assist device (Berlin Heart AG, Berlin, Germany) and a membrane oxygenator (Jostra Quadrox D, Maquet Cardiopulmonary, AG Hirrlingen, Germany) intercalated in the outflow cannula of the left pump. PMID- 19560706 TI - Immune tolerance after total lymphoid irradiation for heart transplantation: immunosuppressant-free survival for 8 years. AB - A 51-year-old African American man underwent orthotopic heart transplantation in 1995 for post-viral cardiomyopathy. Refractory rejection occurred, and he subsequently required total lymphoid irradiation to prevent further rejection. Disseminated Mycobacterium avium complex developed in 2000, and the patient decided to discontinue all drugs after the antibiotics caused intolerable medication side effects. The patient did not subsequently die of rejection, and he was discovered to have profound suppression of several lymphocytes subsets, presumably from the previous total lymphoid irradiation. This induced immunotolerance appears to have enabled his prolonged immunosuppressant-free survival. PMID- 19560707 TI - Regression of cardiac amyloid after autologous stem-cell transplantation. AB - Primary cardiac amyloid has a dismal prognosis and most treatments are experimental with highly variable results. Although autologous stem-cell transplant in conjunction with high-dose chemotherapy has yielded regression of amyloid in other body tissues, the presence of cardiac involvement contraindicates stem-cell transfer due to high treatment mortality. We describe the successful treatment of cardiac amyloid using autologous stem-cell transplantation and the resultant regression of the cardiac amyloid. PMID- 19560708 TI - Pancytopenia with hypercellular bone marrow secondary to cytomegalovirus after cardiac transplant. AB - We describe a case of cytomegalovirus-induced pancytopenia in a heart transplant patient. The interesting finding of a hypercellular bone marrow indicates the possibility of a different mechanism for the pancytopenia other than bone marrow suppression. The patient was treated with ganciclovir, which resulted in the complete resolution of the viremia and pancytopenia. Cytomegalovirus infection should be suspected in heart transplant patients with pancytopenia, regardless of bone marrow results. The pathogenesis of cytomegalovirus-induced pancytopenia is likely multifactorial, with both a central and peripheral effect. PMID- 19560709 TI - Recovery from anthracycline cardiomyopathy--how can it be explained? PMID- 19560710 TI - Re: Lung transplantation in older patients with cystic fibrosis: analysis of UNOS data. PMID- 19560711 TI - Probiotics and the nursing home: should we give bacteria for breakfast? PMID- 19560712 TI - Are you on the health information technology bandwagon? PMID- 19560713 TI - Beyond the medical model: the culture change revolution in long-term care. AB - Culture change in long-term care facilities involves a shift in philosophy and practice from an overemphasis on safety, uniformity, and medical issues toward resident-directed, consumer-driven health promotion and quality of life. Fundamental to this shift is a focus on the importance of the relationships between residents and direct care staff. This review presents and discusses the key elements of culture change, including workforce redesign, resident-centered care, leadership, and the implementation process and evaluation. A case report describes how medical staff can participate in this grassroots movement and help foster the social, cultural, programmatic, and physical changes that can alter the culture of long-term care one home at a time. PMID- 19560714 TI - The development of a mental status vital sign for use across the spectrum of care. PMID- 19560715 TI - Apathy: a common psychiatric syndrome in the elderly. AB - Apathy, or a lack of motivation, has been increasingly recognized as a distinct psychiatric syndrome. Apathy is primarily a dysfunction of the frontal subcortical circuit and is associated with various neuropsychiatric disorders including Alzheimer's disease. Apathy is associated with a number of adverse outcomes, including apparent cognitive impairment, decreased daily function, poor insight into one's own functional and cognitive impairment, and poor outcome from rehabilitation treatment. Furthermore, the degree of caregiver's burden in these patients is significant. This article reviews the definition of apathy, prevalence and associated adverse outcomes, causation, the approach to patients with apathy, and available treatment options with particular attention to studies conducted in a nursing home setting. The purpose of this article is to increase the recognition of apathy by physicians working in the nursing home. PMID- 19560716 TI - Postprandial walking is better for lowering the glycemic effect of dinner than pre-dinner exercise in type 2 diabetic individuals. AB - OBJECTIVES: In prior studies of exercise done before or after breakfast and lunch, postprandial activity generally reduces glycemia more than pre-meal. This study sought to examine the effects of exercise before or after an evening meal. DESIGN: Examined the differing effects of a single bout of pre- or postprandial moderate exercise or no exercise on the glycemic response to an evening (dinner) meal in individuals with type 2 diabetes. SETTING: Community-dwelling participants tested at a research university in Virginia. PARTICIPANTS: Twelve men and women subjects (mean age of 61.4+/-2.7 years) with type 2 diabetes treated with diet and/or oral medications. INTERVENTION: Three trials conducted on separate days consisting of a rest day when subjects consumed a standardized dinner with a moderate glycemic effect and 2 exercise days when they undertook 20 minutes of self-paced treadmill walking immediately before or 15 to 20 minutes after eating. MEASUREMENTS: Blood samples taken every 30 minutes over a 4-hour period and later assayed for plasma glucose; from these data both absolute and relative changes in glucose levels were determined, as well as the total glucose area under the curve (AUC) of the 4-hour testing period. Initial samples were additionally assayed for glycated hemoglobin and lipid levels. RESULTS: Twenty minutes of self-paced walking done shortly after meal consumption resulted in lower plasma glucose levels at the end of exercise compared to values at the same time point when subjects had walked pre-dinner. Total glucose AUC over 4-hours was not significantly different among trials. CONCLUSION: Postprandial walking may be more effective at lowering the glycemic impact of the evening meal in individuals with type 2 diabetes compared with pre-meal or no exercise and may be an effective means to blunt postprandial glycemic excursions. PMID- 19560717 TI - Information technology sophistication in nursing homes. AB - OBJECTIVE: There is growing recognition that a more sophisticated information technology (IT) infrastructure is needed to improve the quality of nursing home care in the United States. The purpose of this study was to explore the concept of IT sophistication in nursing homes considering the level of technological diversity, maturity and level of integration in resident care, clinical support, and administration. METHODS: Twelve IT stakeholders were interviewed from 4 nursing homes considered to have high IT sophistication using focus groups and key informant interviews. Common themes were derived using qualitative analytics and axial coding from field notes collected during interviews and focus groups. RESULTS: Respondents echoed the diversity of the innovative IT systems being implemented; these included resident alerting mechanisms for clinical decision support, enhanced reporting capabilities of patient-provider interactions, remote monitoring, and networking among affiliated providers. DISCUSSION: Nursing home IT is in its early stages of adoption; early adopters are beginning to realize benefits across clinical domains including resident care, clinical support, and administrative activities. The most important thread emerging from these discussions was the need for further interface development between IT systems to enhance integrity and connectivity. CONCLUSION: The study shows that some early adopters of sophisticated IT systems in nursing homes are beginning to achieve added benefit for resident care, clinical support, and administrative activities. PMID- 19560718 TI - Attitudes toward information and communication technology (ICT) in residential aged care in Western Australia. AB - OBJECTIVES: Determine why introduction of health consulting services via Telehealth video conference consultations failed in residential aged care facilities (RACF). DESIGN: Semistructured interview groups and quantitative survey. SETTING: Two participating not-for-profit RACF. PARTICIPANTS: Managers, employed carers, physiotherapist, occupational therapist, registered nurses, and residents from RACF. MEASUREMENTS: A survey initially followed by focus groups that centered on 4 questions. How can computers help improve care? What kind of electronic services and products could help improve care? Who should have access to the technology and why was the technology not used? RESULTS: The survey revealed there was awareness of information and communication technology (ICT) in RACF. However, respondents were uncertain of potential benefits provided to their clients. Only 43% of respondents thought a minority of clients would receive the benefits of ICT use. The focus groups revealed several themes regarding the attitudes toward ICT in RACF. Positive attitudes to ICT included themes of saving time, easier doctor access, cost saving, and improved communications. Negative attitudes included themes of loss of human contact, inadequate training, security barriers, not user friendly, limited ability to comply with suggestions, privacy issues, and capital cost. Residents were also concerned about confidentiality and loss of human interaction with the use of Telehealth in residential aged facilities. CONCLUSIONS: More training for staff is required to enable them to use ICT efficiently. ICT hardware and software at the user interface must be designed to maintain confidentiality with ease of access. Access to Telehealth services should not impede the routine delivery of personal care and human contact for residents. Studies are required as to where human input to residents is unable to be replaced by Telehealth services. PMID- 19560719 TI - Nursing homes as reservoirs of MRSA: myth or reality? AB - OBJECTIVE: To determine the prevalence and risk factors associated with methicillin-resistant Staphylococcus aureus (MRSA) colonization among residents of a long-term care facility (LTCF) and to investigate the association of prior antibiotics use and MRSA colonization. DESIGN: Cross-sectional analysis. SETTING: A large, 320-bed suburban long-term care facility in New York. PARTICIPANTS: A representative sample of 160 nursing home residents. MEASUREMENTS: We obtained nasal swabs to screen for MRSA colonization and reviewed the medical charts for clinical and demographic data. RESULTS: A total of 160 residents participated. MRSA colonization was identified in 44 residents (27.5 %). Only 5 variables were statistically significantly associated with MRSA colonization, namely race, renal insufficiency, increased use of antibiotics, prior MRSA infection during the previous year, and prior hospitalization within 3 years. Sharing a room with a MRSA carrier did not increase the risk for colonization. CONCLUSION: This study found a large reservoir of MRSA within this LTCF population. Nursing home residents with renal insufficiency, prior MRSA infection, prior hospitalization, and higher use of antibiotics were found to be at risk for MRSA colonization . These findings demonstrate that LTCFs need to be proactive in implementing appropriate antibiotics restriction practices and should give high priority to the development of more effective infection control policies. PMID- 19560720 TI - Nursing home policies on items brought in from the outside for facility residents. AB - OBJECTIVE: To identify nursing home standards related to items brought in from the community for residents through a nationwide survey of directors of nursing. Specifically we examined the policies with regard to food, cigarettes, alcoholic beverages, and over-the-counter medications. METHODS: A national survey was distributed online and was completed by 299 directors of nursing of skilled nursing facilities. The directors of nursing were asked about policies regarding whether family and friends of residents are permitted to bring in items such as food, cigarettes, alcohol, and over-the-counter medications. Specifically, questions were related to monitoring, staff involvement, safety precautions, and policy implementation. RESULTS: The results of the survey demonstrated a consistent policy practiced among facilities. Items commonly restricted for all residents included over-the-counter medications, alcohol, and cigarettes. On the other hand, food was significantly less likely to be restricted. CONCLUSION: Despite overall strict policies regarding the monitoring of access to over-the counter medications, alcohol, and cigarettes by nursing home residents, ingestion of outside food remains fairly unrestricted. This is especially concerning given the growing number of residents with end-stage congestive heart failure, diabetes, dysphagia, or food allergies where access to outside food could result in an adverse event. Perhaps, facilities need to identify at-risk residents and better communicate to residents and their families regarding dietary restrictions on outside food. PMID- 19560721 TI - Tenure, certification, and education of nursing home administrators, medical directors, and directors of nursing in for-profit and not-for-profit nursing homes: United States 2004. AB - OBJECTIVES: To understand key characteristics of the leadership team, and to examine if differences in these factors exist between for-profit (FP) and not-for profit (NFP) nursing homes (NHs). DESIGN: Cross sectional. SETTING: US nursing homes. PARTICIPANTS: A nationally representative sample of 1174 US NHs conducted in 2004. INTERVENTION: N/A. MEASUREMENTS: Reported data on tenure, education, and certification of NH administrators (NHAs), medical directors (MeDs), and directors of nursing (DoNs) at FP and NFP facilities. RESULTS: NHAs, MeDs, and DoNs at NFP facilities all had significantly greater tenure at their current facilities compared with their FP counterparts. NHAs and MeDs at NFP facilities were also more likely to have more years of accumulated experience in those roles. MeD certifications differed substantially by specialty, with 23.3%, 37.6%, and 43.5% of MeDs having certification in geriatric, internal, and family medicine, respectively, and about 42% of MeDs were certified by AMDA. However, no differences in MeD certification were observed by facility ownership. Although 68% of all US nursing homes had a MeD who spent 4 or fewer days per month in the facility and only 14% spent 11 days or more per month in the facility, nearly twice as many NFP MeDs spent 11 days or more onsite in the facility compared with FP MeDs. Facility ownership was strongly associated with NHA educational attainment, with a significantly higher proportion of NFP NHAs having master's degrees or higher (41.4% versus 26.6%, P < .0001), and smaller proportions of NFP NHAs having a bachelor's degree or less. CONCLUSION: In 2004, members of the leadership teams of NFP NHs had more favorable profiles for several characteristics related to education and tenure compared with their FP counterparts. More research is needed to understand how variation in leadership skills and capacity affects quality of care and quality of work life outcomes, including the role of FP/NFP differences in explaining differential quality outcomes. PMID- 19560722 TI - Impact of medical director certification on nursing home quality of care. AB - OBJECTIVE: This study tests the research hypothesis that certified medical directors are able to use their training, education, and knowledge to positively influence quality of care in US nursing homes. DESIGN: F-tag numbers were identified within the State Operations Manual that reflect dimensions of quality thought to be impacted by the medical director. A weighting system was developed based on the "scope and severity" level at which the nursing homes were cited for these specific tag numbers. Then homes led by certified medical directors were compared with homes led by medical directors not known to be certified. DATA/PARTICIPANTS: Data were obtained from the Centers for Medicare & Medicaid Services' Online Survey Certification and Reporting database for nursing homes. Homes with a certified medical director (547) were identified from the database of the American Medical Directors Association. MEASUREMENTS: The national survey database was used to compute a "standardized quality score" (zero representing best possible score and 1.0 representing average score) for each home, and the homes with certified medical directors compared with the other homes in the database. Regression analysis was then used to attempt to identify the most important contributors to measured quality score differences between the homes. RESULTS: The standardized quality score of facilities with certified medical directors (n=547) was 0.8958 versus 1.0037 for facilities without certified medical directors (n=15,230) (lower number represents higher quality). When nursing facility characteristics were added to the regression equation, the presence of a certified medical director accounted for up to 15% improvement in quality. CONCLUSIONS: The presence of certified medical directors is an independent predictor of quality in US nursing homes. PMID- 19560724 TI - Natural light exposure improves subjective sleep quality in nursing home residents. PMID- 19560725 TI - Placebo-controlled studies and the need for expanded disclosure rules. PMID- 19560723 TI - Assessing the quality of prescribing and monitoring erythropoiesis-stimulating agents in the nursing home setting. AB - INTRODUCTION: As many as 50% of all nursing home (NH) residents meet the World Health Organization criteria for anemia. The objectives of this study were to determine the prevalence and appropriateness of prescribing and monitoring of erythropoiesis-stimulating agents (ESAs) used to treat anemia in the NH setting. METHODS: Cross-sectional, 1-month study of all NH residents in 4 community-based, university-affiliated NHs between January and February 2008. Residents were included in the analysis if they received at least 1 dose of an ESA during the study duration. Data collected through chart review included basic demographic information, ESA indication, ESA dosage, concurrent administration of iron supplements, hemoglobin (Hgb) monitoring, and blood pressure measurements. RESULTS: A total of 4.5% (22/485) of NH residents received at least 1 dose of an ESA. Residents who received ESAs had a mean age of 80.4 (+/- 14.5) years. Most residents who received ESAs were female (64% [14/22]), white (68% [15/22]), and had a mean weight of 72.0 (+/- 20.84) kg. Only 27% (6/22) of residents were prescribed an ESA for an FDA-approved indication. Darbepoetin alfa was the most commonly prescribed ESA (64% [14/22]) with a mean weekly dose of 70.8 (+/- 68.1) mug, followed by epoetin alfa (37% [8/22]) with a mean weekly dose of 22,625 (+/- 21,232) units. More than one quarter (27% [6/22]) of those who received an ESA had an Hgb value of 12g/dL or more, the maximum recommended threshold for use of these medications. Of the 18 residents who had blood pressure measurements, 11% (2/18) were hypertensive. CONCLUSION: Suboptimal prescribing and monitoring of ESAs were common in the NHs we studied. Future studies are needed to determine if the development and use of computerized decision support systems can improve prescribing and monitoring of ESAs in the NH setting. PMID- 19560726 TI - Aripiprazole for the treatment of agitation in persons with dementia? PMID- 19560729 TI - Chronic kidney disease, anemia, and the association between chronic kidney disease-related anemia and activities of daily living in older nursing home residents by Schnelle et al, February 2009. PMID- 19560730 TI - Predictors of 3-year mortality in institutionalized nonagenarians: the NonaSantfeliu study. PMID- 19560732 TI - Current perspectives on neuro-oncology. PMID- 19560733 TI - Molecular epidemiology of primary brain tumors. AB - Although primary brain tumors (PBTs) are generally considered to be a multifactorial disorder, understanding the genetic basis and etiology of the disease is essential for PBT risk assessment. Understanding of the genetic susceptibility for PBT has come from studies of rare genetic syndromes, linkage analysis, family aggregation, early-onset pediatric cases, and mutagen sensitivity. There are currently no effective markers to assess biological dose of exposures and genetic heterogeneity. The priorities recently recommended by the Brain Tumor Epidemiology Consortium emphasized the need for expanding research in genetics and molecular epidemiology. In this article, we review the literature to identify molecular epidemiologic case-control studies of PBTs that were hypothesis-driven and focused on four hypothesized candidate pathways: DNA repair, cell cycle, metabolism, and inflammation. We summarize the results in terms of genetic associations of single nucleotide polymorphisms of these pathways. We also discuss future research directions based on available evidence and technologies, and conclude that high resolution whole genome approach with significantly large sample size could rapidly advance our understanding of the genetic etiology of PBTs. Literature searches were done on PubMed in March 2009 with the terms glioma, glioblastoma, brain tumor, association, and polymorphism, and we only reviewed English language publications. PMID- 19560734 TI - Molecular epigenetics and genetics in neuro-oncology. AB - Gliomas arise through genetic and epigenetic alterations of normal brain cells, although the exact cell of origin for each glioma subtype is unknown. The alteration-induced changes in gene expression and protein function allow uncontrolled cell division, tumor expansion, and infiltration into surrounding normal brain parenchyma. The genetic and epigenetic alterations are tumor subtype and tumor-grade specific. Particular alterations predict tumor aggressiveness, tumor response to therapy, and patient survival. Genetic alterations include deletion, gain, amplification, mutation, and translocation, which result in oncogene activation and tumor suppressor gene inactivation, or in some instances the alterations may simply be a consequence of tumorigenesis. Epigenetic alterations in brain tumors include CpG island hypermethylation associated with tumor suppressor gene silencing, gene-specific hypomethylation associated with aberrant gene activation, and genome-wide hypomethylation potentially leading to loss of imprinting, chromosomal instability, and cellular hyperproliferation. Other epigenetic alterations, such as changes in the position of histone variants and changes in histone modifications are also likely to be important in the molecular pathology of brain tumors. Given that histone deacetylases are targets for drugs that are already in clinical trial, surprisingly little is known about histone acetylation in primary brain tumors. Although a majority of epigenetic alterations are independent of genetic alterations, there is interaction on specific genes, signaling pathways and within chromosomal domains. Next generation sequencing technology is now the method of choice for genomic and epigenome profiling, allowing more comprehensive understanding of genetic and epigenetic contributions to tumorigenesis in the brain. PMID- 19560735 TI - Biology of angiogenesis and invasion in glioma. AB - Treatment of adult brain tumors, in particular glioblastoma, remains a significant clinical challenge, despite modest advances in surgical technique, radiation, and chemotherapeutics. The formation of abnormal, dysfunctional tumor vasculature and glioma cell invasion along white matter tracts are believed to be major components of the inability to treat these tumors effectively. Recent insight into the fundamental processes governing glioma angiogenesis and invasion provide a renewed hope for development of novel strategies aimed at reducing the morbidity of this uniformly fatal disease. In this review, we discuss background biology of the blood brain barrier and its pertinence to blood vessel formation and tumor invasion. We will then focus our attention on the biology of glioma angiogenesis and invasion, and the key mediators of these processes. Last, we will briefly discuss recent and ongoing clinical trials targeting mediators of angiogenesis or invasion in glioma patients. The findings provide a renewed hope for those endeavoring to improve treatment of patients with glioma by providing a novel set of rational targets for translational drug discovery. PMID- 19560736 TI - Novel treatment strategies for malignant gliomas using neural stem cells. AB - Recent studies in stem cell biology have refined our understanding of the origin and progression of cancer. Identification and characterization of endogenous neural stem cells (NSCs), especially those in the adult human brain, have inspired new ideas for selectively targeting and destroying malignant gliomas. Gliomas consist of a heterogeneous population of cells, and some of these cells have characteristics of cancer stem cells. These brain tumor stem cells (BTSCs) share certain characteristics with normal NSCs. It is still unclear, however, whether malignant gliomas in human patients originate from these aberrant BTSCs. Nonetheless, the cellular and molecular similarities between BTSCs and normal NSCs suggest a common research landscape underlying both normal and cancer stem cell biology, wherein findings of one field are relevant to the other. Furthermore, the natural tropism of NSCs to gliomas has generated the idea that modified NSCs can deliver modified genes to selectively destroy malignant brain tumor cells, and even BTSCs, while leaving healthy surrounding neurons intact. These studies and others on the basic biology of both BTSCs and NSCs will be crucial to expanding our treatment strategies for malignant gliomas. PMID- 19560737 TI - Neuroimaging in neuro-oncology. AB - Neuroimaging of brain tumors has evolved from a strictly morphology-based discipline to one that encompasses function, physiology, and anatomy. This review outlines the current imaging standard for patients with brain tumor and summarizes the latest advances in physiology-based imaging methods that complement traditional brain tumor imaging protocols. Emphasis is on the strength and limitations of the current imaging standards and on an overview of several advanced imaging methods including diffusion-weighted magnetic resonance imaging (MRI), perfusion MRI, and proton magnetic resonance spectroscopic (MRS) imaging. Basic physical principles behind each imaging method are briefly presented, along with a more in-depth discussion of clinical application and potential pitfalls of each technique. PMID- 19560738 TI - Operative techniques for gliomas and the value of extent of resection. AB - Refinement of neurosurgical technique has enabled safer operations with more aggressive outcomes. One cornerstone of modern-day practice is the utilization of intraoperative stimulation mapping. In addition to identifying critical motor pathways, this technique can be adapted to reliably identify language pathways. Given the individual variability of cortical language localization, such awake language mapping is essential to minimize language deficits following tumor resection. Our experience suggests that cortical language mapping is a safe and efficient adjunct to optimize tumor resection while preserving essential language sites, even in the setting of negative mapping data. However, the value of maximizing glioma resections remains surprisingly unclear, as there is no general consensus in the literature regarding the efficacy of extent of glioma resection in improving patient outcome. While the importance of resection in obtaining tissue diagnosis and alleviating symptoms is clear, a lack of Class I evidence prevents similar certainty in assessing the influence of extent of resection. Beyond an analysis of modern intraoperative mapping techniques, we examine every major clinical publication since 1990 on the role of extent of resection in glioma outcome. The mounting evidence suggests that, despite persistent limitations in the quality of available studies, a more extensive surgical resection is associated with longer life expectancy for both low-grade and high grade gliomas. PMID- 19560739 TI - Radiation techniques in neuro-oncology. AB - Radiation therapy plays a critical role in the management of tumors of the brain. A variety of radiotherapy techniques have been used to treat these tumors. This review describes both classic and more recent and advanced techniques available to manage these tumors. Included is a discussion of standard two- and three dimensional radiation, as well as intensity-modulated radiotherapy, image-guided radiation therapy, stereotactic radiosurgery, and heavy particles. PMID- 19560740 TI - Targeted therapy for malignant glioma patients: lessons learned and the road ahead. AB - Molecularly targeted therapies are transforming the care of patients with malignant gliomas, including glioblastoma, the most common malignant primary brain tumor of adults. With an arsenal of small molecule inhibitors and antibodies that target key components of the signal transduction machinery that are commonly activated in gliomas, neuro-oncologists and neurosurgeons are poised to transform the care of these patients. Nonetheless, successful application of targeted therapies remains a challenge. Strategies are lacking for directing kinase inhibitor or other pathway-specific therapies to individual patients most likely to benefit. In addition, response to targeted agents is determined not only by the presence of the key mutant kinases, but also by other critical changes in the molecular circuitry of cancer cells, such as loss of key tumor suppressor proteins, the selection for kinase-resistant mutants, and the deregulation of feedback loops. Understanding these signaling networks, and studying them in patients, will be critical for developing rational combination therapies to suppress resistance for malignant glioma patients. Here we review the current status of molecular targeted therapies for malignant gliomas. We focus initially on identifying some of the insights gained to date from targeting the EGFR/PI3K/Akt/mTOR signaling pathway in patients and on how this has led toward a reconceptualization of some of the challenges and directions for targeted treatment. We describe how advances from the world of genomics have the potential to transform our approaches toward targeted therapy, and describe how a deeper understanding of the complex nature of cancer, and its adeptness at rewiring molecular circuitry to evade targeted agents, has raised new challenges and identified new leads. PMID- 19560741 TI - Antiangiogenic strategies for treatment of malignant gliomas. AB - Numerous antiangiogenic agents with diverse mechanisms of action are currently under investigation for the treatment of patients with glioblastoma (GBM), a diagnosis that continues to carry a poor prognosis despite maximal conventional therapy. Early clinical trials suggest that antiangiogenic drugs, which target the blood vessels of these highly angiogenic tumors, may have clinical benefit in GBM patients. Antiangiogenic agents have potent antiedema and steroid-sparing effects in patients, and emerging data suggest that these drugs may modestly improve progression-free survival. Although these early results are encouraging, several issues arise regarding the use and efficacy of these agents. Interpretation of the radiographic changes that occur after treatment with antiangiogenic agents presents a major challenge. Still lacking are reliable radiographic and biologic markers that can predict which patients will benefit from treatment and that accurately indicate response and progression during therapy. In addition, most patients treated with antiangiogenic drugs eventually progress, and the mechanisms by which tumors escape from therapy are only beginning to be understood. Larger prospective trials that incorporate correlative biomarker studies will be required to address these challenges. Here, we summarize the clinical experience with antiangiogenic therapy in patients with malignant gliomas (MG), review the major issues concerning the use and development of these agents, and discuss strategies that may build upon the initial gains observed with antiangiogenic agents. PMID- 19560743 TI - Novel drug delivery strategies in neuro-oncology. AB - Treatment of malignant gliomas represents one of the most formidable challenges in oncology. Despite treatment with surgery, radiation therapy, and chemotherapy, the prognosis remains poor, particularly for glioblastoma, which has a median survival of 12 to 15 months. An important impediment to finding effective treatments for malignant gliomas is the presence of the blood brain barrier, which serves to prevent delivery of potentially active therapeutic compounds. Multiple efforts are focused on developing strategies to effectively deliver active drugs to brain tumor cells. Blood brain barrier disruption and convection enhanced delivery have emerged as leading investigational delivery techniques for the treatment of malignant brain tumors. Clinical trials using these methods have been completed, with mixed results, and several more are being initiated. In this review, we describe the clinically available methods used to circumvent the blood brain barrier and summarize the results to date of ongoing and completed clinical trials. PMID- 19560742 TI - Toward effective immunotherapy for the treatment of malignant brain tumors. AB - The immunologic treatment of cancer has long been heralded as a targeted molecular therapeutic with the promise of eradicating tumor cells with minimal damage to surrounding normal tissues. However, a demonstrative example of the efficacy of immunotherapy in modulating cancer progression is still lacking for most human cancers. Recent breakthroughs in our understanding of the mechanisms leading to full T-cell activation, and recognition of the importance of overcoming tumor-induced immunosuppressive mechanisms, have shed new light on how to generate effective anti-tumor immune responses in humans, and sparked a renewed and enthusiastic effort to realize the full potential of cancer immunotherapy. The immunologic treatment of invasive malignant brain tumors has not escaped this re-invigorated endeavor, and promising therapies are currently under active investigation in dozens of clinical trials at several institutions worldwide. This review will focus on some of the most important breakthroughs in our understanding of how to generate potent anti-tumor immune responses, and some of the clear challenges that lie ahead in achieving effective immunotherapy for the majority of patients with malignant brain tumors. A review of immunotherapeutic strategies currently under clinical evaluation, as well as an outline of promising novel approaches on the horizon, is included to provide perspective on the active and stalwart progress toward effective immunotherapy for the treatment of malignant brain tumors. PMID- 19560744 TI - Gene- and viral-based therapies for brain tumors. AB - Advances in understanding and controlling genes and their expression have set the stage to alter genetic material to fight or prevent disease with brain tumors being among one of the first human malignancies to be targeted by gene therapy. All proteins are coded for by DNA and most neoplastic diseases ultimately result from the expression or lack thereof with one or more proteins (e.g., coded by oncogenes or tumor suppressor genes, respectively). In theory, therefore, diseases could be treated by expression of the appropriate protein in the affected cells. Gene therapy is an experimental treatment that involves introducing genetic material (DNA or RNA) into cells, and it has made important advances in the past decade. Within this short time span, it has moved from the conceptual laboratory research stage to clinical translational trials for brain tumors. The most efficient approaches for gene delivery are based on viral vectors, which have been proven relatively safe in the CNS, despite occasional cases of morbidity and death in non-neurosurgical trials. However, the human response to various viral vectors can not be predicted in a reliable manner from animal experimentation, nor can size, consistency, and extent of experimental brain tumors in mouse models reflect the large, necrotic, infiltrative nature of malignant gliomas. Furthermore, the problem of delivering genetic vectors into solid brain tumors and the efficiency in situ gene transfer remains one of the most significant hurdles in gene therapy. PMID- 19560745 TI - Oncolytic viral therapy of malignant glioma. AB - Novel approaches to treatment of malignant glioma, the most frequently occurring primary brain tumor, have included the use of a wide range of oncolytic viral vectors. These vectors, either naturally tumor-selective, or engineered as such, have shown promise in the handful of phase I and phase II clinical trials conducted in recent years. The strategies developed for each of the different viruses currently being studied and the history of their development are summarized here. In addition, the results of clinical trials in patients and their implication for future trials are also discussed. PMID- 19560747 TI - Biology and treatment of primary central nervous system lymphoma. AB - Primary central nervous system lymphoma (PCNSL) is a rare variant of extranodal non-Hodgkin lymphoma that is restricted in distribution to the brain, leptomeninges, spinal cord, and intraocular compartments. Although PCNSL shares overlapping features with systemic lymphoma, recent studies also reveal a unique pattern of gene and protein expression in PCNSL. These findings have yielded new insights into the pathophysiology of the disease, as well as the identification of novel prognostic biomarkers. Immune system compromise, such as is seen in acquired immune deficiency syndrome (AIDS), is the best established known risk factor for PCNSL. Like other lesions of the brain, meninges, and eye, the presenting symptoms associated with PCNSL typically include focal neurological deficits related to the site of disease or more global consequences of increased intracranial pressure. Diagnosis of PCNSL typically includes gadolinium-enhanced MRI and pathologic tissue analysis, as well as additional studies aimed at excluding concurrent systemic disease. PCNSL typically has a worse overall prognosis than systemic lymphoma. High-dose chemotherapy, particularly with methotrexate-based regimens, is the backbone of therapy for most patients, and chemotherapy is associated with much lower rates of treatment-related morbidity and mortality than whole-brain irradiation. Autologous stem cell transplantation is an emerging treatment modality, particularly in younger patients with relapsed disease, but high rates of treatment-related mortality are observed in older patients. Immunotherapy, including treatment with intrathecal rituximab, is another area of active research that may have promise in refractory or relapsed disease. Treatment options for intraocular lymphoma parallel those for PCNSL elsewhere in the brain: systemic chemotherapy, radiation, and local delivery of cytotoxic and immunologically active agents such as anti-CD20 antibody. PMID- 19560748 TI - Current management of metastatic brain disease. AB - Brain metastases are the most common intracranial tumor in adults. The incidence of metastases is thought to be rising due to better detection and treatment of systemic malignancy. More widespread use and improved quality of MRI may lead to early detection of brain metastases. Available evidence suggests that survival is longer and quality of life improved if brain metastases are treated aggressively. This article reviews current therapeutic management used for brain metastases. To select the appropriate therapy, the physician must consider the extent of the systemic disease, primary histology, and patient age and performance status, as well as the number, size, and location of the brain metastases. Available treatment options include whole-brain radiotherapy (WBRT), stereotactic radiosurgery (SRS), surgery, and chemotherapy. Multidisciplinary approaches such as the combination of WBRT with SRS or surgery have shown superior results in terms of survival time, neurocognitive function, and quality of life. The utility and optimal use of chemotherapy and radiosensitizing agents is less clear. It is hoped that further advances and multidisciplinary approaches currently under study will result in improved patient outcomes. PMID- 19560746 TI - Pediatric brain tumors: current treatment strategies and future therapeutic approaches. AB - Pediatric CNS tumors are the most common solid tumors of childhood and the second most common cancer after hematological malignancies accounting for approximate 20 to 25% of all primary pediatric tumors. With over 3,000 new cases per year in the United States, childhood CNS tumors are the leading cause of death related to cancer in this population. The prognosis for these patients has improved over the last few decades, but current therapies continue to carry a high risk of significant side effects, especially for the very young. Currently a combination of surgery, radiation, and chemotherapy is often used in children greater than 3 years of age. This article will outline current and future therapeutic strategies for the most common pediatric CNS tumors, including primitive neuroectodermal tumors such as medulloblastoma, as well as astrocytomas and ependymomas. PMID- 19560749 TI - Long-term medication adherence after myocardial infarction: experience of a community. AB - BACKGROUND: Adherence to evidence-based medications after myocardial infarction is associated with improved outcomes. However, long-term data on factors affecting medication adherence after myocardial infarction are lacking. METHODS: Olmsted County residents hospitalized with myocardial infarction from 1997-2006 were identified. Adherence to HMG-CoA reductase inhibitors (statins), beta blockers, angiotensin-converting enzyme inhibitors, and angiotensin II receptor blockers, were examined. Cox proportional hazard regression was used to determine the factors associated with medication adherence over time. RESULTS: Among 292 subjects with incident myocardial infarction (63% men, mean age 65 years), patients were followed for an average of 52+/-31 months. Adherence to guideline recommended medications decreased over time, with 3-year medication continuation rates of 44%, 48%, and 43% for statins, beta-blockers, and angiotensin-converting enzyme inhibitors/angiotensin II receptor blockers, respectively. Enrollment in a cardiac rehabilitation program was associated with an improved likelihood of continuing medications, with adjusted hazard ratio (95% confidence interval) for discontinuation of statins and beta-blockers among cardiac rehabilitation participants of 0.66 (0.45-0.92) and 0.70 (0.49-0.98), respectively. Smoking at the time of myocardial infarction was associated with a decreased likelihood of continuing medications, although results did not reach statistical significance. There were no observed associations between demographic characteristics, clinical characteristics of the myocardial infarction, and medication adherence. CONCLUSIONS: After myocardial infarction, a large proportion of patients discontinue use of medications over time. Enrollment in cardiac rehabilitation after myocardial infarction is associated with improved medication adherence. PMID- 19560752 TI - Single-crystal and powder X-ray diffraction and solid-state 13C NMR of p nitrophenyl glycopyranosides, the derivatives of D-galactose, D-glucose, and D mannose. AB - The X-ray diffraction patterns, (13)C CP MAS NMR spectra, and powder X-ray diffraction analyses were obtained for selected p-nitrophenyl glycosides: alpha- and beta-D-galactopyranosides (1 and 2), alpha- and beta-D-glucopyranosides (3 and 4), and alpha- and beta-D-mannopyranosides (5 and 6). In X-ray diffraction analysis of 1 and 2, characteristic shortening and lengthening of selected bonds were observed in the molecules of 1 due to anomeric effect, and in the crystal lattice of 1 and 2, hydrogen bonds of complex network were detected. In the crystal asymmetric unit of 1 there were two independent molecules, whereas in 2 there was one molecule. For 1 and 3-6 the number of resonances in solid-state (13)C NMR spectra exceeded the number of the carbon atoms in the molecules, while for 2 there were distinct singlet resonances in its solid-state NMR spectrum. Furthermore, the powder X-ray diffraction (PXRD) performed for 1-3 and 5 revealed that 1, 3, and 5 existed as single polymorphs proving that the doublets observed in appropriate solid-state NMR spectra were connected with two non-equivalent molecules in the crystal asymmetric unit. On the other hand 2 existed as a mixture of two polymorphs, one of them was almost in agreement with the calculated pattern obtained from XRD (the difference in volumes of the unit cells), and the subsequent unknown polymorph existed in small amounts and therefore it was not observed in solid-state NMR measurements. PMID- 19560750 TI - [Endostenting using a Y-stent for hematoma after thoracic aorta surgery]. AB - OBJECTIVE: To describe the diagnostic and therapeutic management of extrinsic tracheobronchial compression after thoracic aorta surgery. PATIENT AND METHODS: We report the case of a female patient with Marfan syndrome. RESULTS: A 27-year old woman with Marfan syndrome presented respiratory distress after type III dissecting aneurysm of the descending aorta from extrinsic compression (hematoma) of the lower third of the trachea and the left bronchial stump. We placed a Y stent (Y-Tracheobronxane, Novatech SA, France), thus restoring a satisfactory channel. A few days later, the patient developed right congestive heart failure from compression of the left pulmonary artery secondary to the progression of the hematoma. Medical treatment for this cardiological complication and the progressive resorption of the hematoma made it possible to remove the stent after 21 days with no dyspneic recurrence. CONCLUSION: Placing a Y-stent is possible and effective and has few iatrogenic consequences in cases of extrinsic tracheobronchial compression from a hematoma occurring during heavy thoracic vascular surgery. With surgical revision impossible given the risks, stent placement allows progressive resorption of the hematoma, thus ending the compression of the bronchial tree. PMID- 19560751 TI - Cyclic stretching force induces apoptosis in human periodontal ligament cells via caspase-9. AB - The response of periodontal ligament (PDL) cells to mechanical stimulation is important in the periodontal tissue remodelling. Our previous study showed that cyclic stretching force on PDL cells induced early apoptosis. However, the mechanism of stretching force-induced cell death is unclear. In the present study, we examined whether PDL cells undergo apoptosis by stretching force using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labellling method (TUNEL) and investigated the mechanism by which cyclic stretching force initiated apoptosis. We found that PDL cells became aligned regularly and the number of apoptotic cells increased significantly in a time-and force-dependent manner after the application of cyclic stretching force. Caspase 3 activity increased in proportion to the magnitude of the stretching force, and this effect was reduced significantly by a caspase-9 inhibitor, whereas a caspase 8 inhibitor had no such effect. We therefore concluded that the in vitro application of cyclic stretching force can induce apoptosis in PDL cells by activating the caspase-3 via the caspase-9 signalling cascade. Our findings may provide a novel insight into the mechanism of apoptosis induced by stretching force in PDL cells. PMID- 19560754 TI - [Influence of bariatric surgery on the non-alcoholic liver steatosis. A histological evaluation]. AB - INTRODUCTION: Non-alcoholic fatty liver (NAFL) is highly prevalent in obese patients. The aim of this study is to look at the development of the histological lesions in these patients that we treated using biliopancreatic diversion. MATERIAL AND METHODS: A liver biopsy was performed on 76 obese patients who were operated on using bariatric surgery. Another liver biopsy was performed on 39 of them between 12 and 24 months after the surgery. The clinical and analytical variables at the time of the surgery, and at 18 months were analysed. RESULTS: A total of 67 patients (88.1%) had NAFL at the time of the operation. Simple steatosis was seen in 41 (61.2%) patients and 26 (38.8%) had non-alcoholic steatohepatitis (NASH). An improvement was seen in both the associated pathology and the analytical evaluations. A significant improvement was observed in the degree of steatosis as well as the NASH in the second biopsy performed on 39 patients. There were no deteriorations in the lesions in any of the patients. CONCLUSIONS: There is a high prevalence of NAFL among obese patients. Bariatric surgery (Scopinaro) leads to weight loss which is associated to an improvement in the pathology associated with obesity, as well as a significant decrease in the liver function values. After weight loss, almost all our patients showed an improvement in the histological hepatic lesions, and in particular, that NASH disappeared in 85% of them. PMID- 19560753 TI - Heat shock protein 90 regulates the stability of MEKK3 in HEK293 cells. AB - Heat shock protein 90 (Hsp90) is a molecular chaperone required for the conformational maturation and function of certain signaling proteins. Hsp90 inhibitors cause the inactivation, destabilization and eventual degradation of Hsp90 client proteins through occupying the ATP/ADP binding pocket of Hsp90. In the present study, we found that Hsp90 interacted with MEKK3 in HEK293 cells. Hsp90 inhibitors reduced the level of endogenous MEKK3 in time- and dose dependent manners, and this decrease was reversed by Hsp90 overexpression. In addition, Hsp90 RNAi destabilized MEKK3. A selective inhibitor of Hsp90, geldanamycin (GA), shortened MEKK3 half-life, and induced ubiquitination and proteasomal degradation of MEKK3. These results strongly suggested that Hsp90 could work as the molecular chaperone of MEKK3. PMID- 19560756 TI - Fewer medications for in vitro fertilization can be better: thinking outside the box. AB - Minimal stimulation or mild stimulation protocols for in vitro fertilization offer many advantages for all patients, but especially for low and high responders. In the current practice of reducing the number of embryos transferred to a maximum of two for a significant number of patients, using aggressive and high-dose gonadotropins to obtain a large, and sometimes excessive, number of oocytes should be questioned. PMID- 19560755 TI - Leveling the playing field for grandfather's sperm. PMID- 19560757 TI - Fertilization rate is an independent predictor of implantation rate. AB - OBJECTIVE: To determine whether fertilization rate serves as a biological assay, reflects oocyte quality, and may be used to help predict patient implantation rate. DESIGN: Retrospective cohort study. SETTING: Academic center. PATIENT(S): Couples undergoing 3603 in vitro fertilization (IVF) cycles from 2001 to 2007. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): We compared the implantation rate among cycles with high versus low fertilization rate. Univariate analyses were performed to determine the association of implantation rate with potential confounding variables: age, day-3 follicle-stimulating hormone level, day-3 estradiol level, antral follicle count, oocyte number, cycle attempts, embryo grading, and number of embryos transferred. Multivariate analysis was then performed to determine whether the fertilization rate remained an independent predictor. RESULT(S): Cutoffs for fertilization rate were 50% for intracytoplasmic sperm injection (ICSI) and 75% for conventional insemination. Higher ICSI fertilization was statistically significantly associated with the implantation rate (25.2% vs. 17.8 %). After adjusting for variables associated with implantation rate, fertilization rate for ICSI remained a strong independent predictor of implantation. Higher conventional insemination fertilization was statistically significantly associated with implantation (32.1% vs. 25.7%) and remained a statistically significant predictor after adjustment. CONCLUSION(S): Fertilization is a strong, independent predictor of implantation rate and may be useful in modeling to guide decision making for the number of embryos to transfer. PMID- 19560758 TI - Effects of follicular phase and oocyte-cumulus complexes quality on the protein profile and in vitro oocyte meiosis competence in Cebus apella. AB - OBJECTIVE: To study the protein profile of oocytes and cumulus cells from different sized follicles throughout the follicular phase and to asses the ability of oocytes to progress from the dictyate to metaphase II (MII) stage. DESIGN: Animal model study. SETTING: Five academic basic research laboratories and the National Primate Centre. ANIMAL(S): Eleven normal, cycling capuchin monkey (Cebus apella) females. INTERVENTION(S): Cumulus-oocyte complexes and denuded oocytes were recovered by antral follicle aspiration. MAIN OUTCOME MEASURE(S): Protein profile analysis of denuded or intact oocytes. RESULT(S): The protein profiles of 25 denuded or intact oocytes recovered on days 5 (six denuded, five intact), 7 (four denuded, four intact), or 9 (one denuded, five intact) of the menstrual cycle were analyzed; in a second experiment, 40 intact oocytes were cultured for 24 (n = 20) or 36 hours (n = 20). The oocytes were denuded, fixed, stained, and microscopically examined to reveal the meiotic stage. The protein profile in each compartment within the cumulus-oocyte complex varied along the follicular development with a predominance of low-molecular weight proteins in both oocyte and cumulus cells at final stages. No differences were found in the protein profile among oocytes pertaining to different sized follicles that were in the same day of the follicular phase. Oocyte MII competence was achieved only after incubation for 36 hours, and the highest maturation rate occurred in those becoming from dominant follicles. CONCLUSION(S): Our study shows, for the first time in a New World primate species, that the proteins contained in oocytes and cumulus cells reach an identical profile in the late follicular phase. This phenomenon could be related to the oocyte's ability to progress to the MII stage. PMID- 19560759 TI - Does dilation and curettage versus expectant management for spontaneous abortion in patients undergoing in vitro fertilization affect subsequent endometrial development? AB - In in vitro fertilization patients, treatment of spontaneous abortion with dilation and curettage (D&C) versus expectant management has no long-term effect on subsequent endometrial development, as measured by change in endometrial thickness. A transient reduction in endometrial thickness was found within the first 6 months after D&C, which is a novel finding, but it is likely to have little or no effect on pregnancy rates given the small absolute effect on endometrial thickness. PMID- 19560760 TI - Higher hospital volume is associated with lower mortality in acute nonvariceal upper-GI hemorrhage. AB - BACKGROUND: Acute nonvariceal upper-GI hemorrhage (NVUGIH) is associated with significant morbidity and mortality. OBJECTIVE: To examine the relationship between hospital volume and outcomes of NVUGIH. DESIGN: A cross-sectional study. SETTING: Participating hospitals from the Nationwide Inpatient Sample 2004. PATIENTS: All discharged patients with a primary discharge diagnosis of NVUGIH based on the International Classification of Diseases, Clinical Modification, ninth edition codes. INTERVENTIONS: Patients were divided into 3 groups based on discharge from hospitals with annual discharge volumes of 1 to 125 (low), 126 to 250 (medium), and >250 (high). MAIN OUTCOME MEASUREMENTS: In-hospital mortality, length of stay, and hospitalization charges. RESULTS: The study included a total of 135,366, 132,746, and 123,007 discharges with NVUGIH occurred from low-volume, medium-volume, and high-volume hospitals, respectively. On multivariate analysis, when adjusting for age, comorbidity, and the presence of complications, patients at high-volume hospitals had significantly lower in-hospital mortality (odds ratio [OR] 0.85 [95% CI, 0.74-0.98]) than patients at low-volume hospitals. Patients at high-volume hospitals were also more likely to undergo upper-GI endoscopy (OR 1.52 [95% CI, 1.36-1.69]) or early endoscopy within 1 day of hospitalization compared with low-volume hospitals (60.5% vs 53.8%, adjusted OR 1.28 [95% CI, 1.02-1.61]). Undergoing endoscopy within day 1 was associated with shorter hospital stays (-1.08 days [95% CI, -1.24 to -0.92 days]) and lower hospitalization charges (-$1958 [95% CI, -$3227 to -$688]). LIMITATIONS: The study was based on an administrative data set. CONCLUSIONS: Higher hospital volume is associated with lower mortality and with higher rates of endoscopy and endoscopic intervention in patients with NVUGIH. PMID- 19560761 TI - Endoscopic resection of distal bile duct mass (with video). PMID- 19560762 TI - Endocytoscopy for the detection of microstructural features in adult patients with celiac sprue: a prospective, blinded endocytoscopy-conventional histology correlation study. AB - BACKGROUND: Endocytoscopy (EC) is a novel technique that allows magnified live inspection of the intestinal mucosa. OBJECTIVE: To evaluate EC for the detection of key pathological findings in patients with celiac sprue. DESIGN: A total of 166 EC recordings were prospectively acquired. Matched videos, images, and biopsy specimens were obtained by duodenal argon beamer labeling of the respective sites. SETTING: Academic tertiary referral center. PATIENTS: Forty patients (mean age 51.5 years, 70% women) with established (n = 32) or suspected (n = 8) celiac disease (CD). INTERVENTIONS: A validated scoring system (Marsh classification) was used to assess disease activity. EC criteria were independently evaluated by 2 gastroenterologists and 1 pathologist. MAIN OUTCOME MEASUREMENTS: The primary endpoint was to examine EC correlation with conventional CD histology. RESULTS: Of 166 duodenal biopsy sites, 23% were classified as Marsh III (moderate to severe), 10% as Marsh I (mild), and 67% as Marsh 0 (normal). Using the 450x magnification, we found that identification of crypts was diagnostic for celiac pathology. Four criteria were significant predictors of Marsh III pathology when adjusted by multivariate analysis: low number of villi per visual field (<3; odds ratio [OR] 9.1; 95% CI, 1.3-62.0), confluence of villi (OR 37.1; 95% CI, 1.3 1021.2), irregular epithelial lining (OR 10.9; 95% CI, 2.5-46.7), and inability to delineate loop capillaries (OR 14.9; 95% CI, 3.3-67.0). None was a good predictor of Marsh I pathology. LIMITATIONS: Single-center experience. No prospective validation of the criteria in an independent patient population. CONCLUSIONS: EC at 450x magnification accurately identifies mucosal histopathology of advanced CD, but not early morphological changes. PMID- 19560763 TI - Mechanisms of bariatric surgery and implications for the development of endoluminal therapies for obesity. PMID- 19560764 TI - Double-guidewire technique for difficult bile duct cannulation: a multicenter randomized, controlled trial. AB - BACKGROUND: ERCP can be associated with serious complications. Difficulty in common bile duct (CBD) cannulation is one of the main risk factors for post-ERCP pancreatitis. The double-guidewire technique (DGT) has been considered a promising alternative approach in difficult cannulation situations. OBJECTIVE: To compare the performance of DGT with the standard cannulation technique (SCT) in patients in whom CBD cannulation is difficult to perform. DESIGN: Multicenter randomized, controlled trial. SETTING: Six tertiary referral centers. PATIENTS: A total of 188 patients with difficult CBD cannulation defined by completion of 5 unsuccessful cannulation attempts were enrolled. INTERVENTIONS: Ninety-seven patients were assigned to the DGT group and 91 to the SCT group. Both techniques were compared for an extra 10 cannulation attempts. MAIN OUTCOME MEASUREMENTS: CBD cannulation rate, number of attempts required to cannulate, and ERCP-related complications. RESULTS: Successful CBD cannulation was achieved in 46 of 97 (47%) patients in the DGT group compared with 51 of 91 (56%) in the SCT group (OR 0.85; 95% CI, 0.64-1.12). The median number of attempts required for each group was 9 and 7, respectively (P = .128). The incidence of post-ERCP pancreatitis was 17% in the DGT group and 8% in the SCT group (OR 2.13; 95% CI, 0.89-5.05). LIMITATIONS: Reduced number of enrolled subjects and a lack of detailed information regarding the number and extent of pancreatic duct contrast injections. CONCLUSIONS: In patients with difficult CBD cannulation, DGT was not superior to SCT in achieving CBD cannulation. DGT might be associated with a higher risk of post-ERCP pancreatitis. PMID- 19560765 TI - Impact of obesity on endoscopy. PMID- 19560766 TI - Natural orifice versus conventional laparoscopic distal pancreatectomy in a porcine model: a randomized, controlled trial. AB - BACKGROUND: Natural orifice transluminal endoscopic surgery (NOTES) research has primarily involved case series reports of low-risk procedures. Distal pancreatectomy has significant postoperative morbidity and would permit rigorous examination in a controlled trial setting. OBJECTIVE: To compare endoscopic transgastric distal pancreatectomy (ETDP) and laparoscopic distal pancreatectomy (LDP). DESIGN: Prospective, randomized, controlled trial. SETTING: Academic hospital. SUBJECTS: Forty-one swine, 28 block randomized. INTERVENTIONS: LDP was performed with 3 trocars and stapled transection of the pancreas. ETDP was performed via a gastrotomy, with 1 trocar for visualization, by using endoloop placement, snare transection, and purse-string gastrotomy closure. MAIN OUTCOME MEASUREMENTS: Clinical examination, CT, serum chemistries, necropsy, peritoneal fluid analysis, and histologic examination. RESULTS: Swine were survived for 8 days. The procedure time for ETDP was significantly greater than for LDP (1:52 vs 0:33 [hours:minutes]; P = .00). Pancreatic specimen weight was similar (4.1 g vs 5.5 g; P = .108). Postoperatively, 26 of 28 animals thrived. In the LDP group, 1 death caused by pancreatic leak and renal failure occurred on day 1. In the ETDP group, 1 death caused by pneumothorax occurred intraoperatively. The necropsy, CT, and histologic examinations revealed focal resection-margin necrosis in 3 to 7 swine in the ETDP group with no proximal necrosis or pancreatitis. The groups were equivalent clinically, by survival, and by serum and peritoneal fluid analysis. The gastrotomy closure was associated with small serosal adhesions, but no gross abscess or necrosis. LIMITATION: Animal study. CONCLUSIONS: In the largest controlled trial of NOTES orifice surgery to date, there was no clinical or survival difference between NOTES and laparoscopic approaches. PMID- 19560767 TI - Esophageal ulceration: a complication of radiofrequency ablation treatment of atrial fibrillation. PMID- 19560769 TI - Actions needed to improve maternal health. AB - The health of mothers and their children is of critical importance, both as a reflection of the current health status of a large segment of the world's population and as a predictor of the health of the next generation. A range of indicators of maternal and neonatal health exist-those primarily affecting pregnant and postpartum women, and those affecting the health and survival of infants. Pregnancy outcome may be affected by toxicant exposure, maternal habits, occupational hazards, psychosocial factors, socioeconomic status, racial disparity, chronic stress, and infections. An increase in obstetric pathologies related to lifestyle, environment, aging, and diet has been seen in Western countries. Large segments of the population are obese and this factor is associated with a great number of adverse reproductive health outcomes. In other countries, the most important objective is to reduce the incidence of infectious diseases and their transmission from mother to fetus. AIDS remains the leading cause of death of children worldwide. PMID- 19560768 TI - Single-operator EUS-guided cholangiopancreatography for difficult pancreaticobiliary access (with video). AB - BACKGROUND: When conventional ERCP methods fail because of periampullary or ductal obstruction, EUS-guided cholangiopancreatography (EUS-CP) may aid in pancreaticobiliary access. OBJECTIVE: To report our experience when using single operator EUS-CP. SETTING: An academic tertiary-referral center. METHODS: Consecutive patients undergoing EUS-CP were prospectively identified. These patients had undergone failed attempt(s) at therapeutic ERCP. A data sheet was used to record indications, reasons for failed ERCP, EUS-CP visualization of the duct of interest, transpapillary or transenteric intervention, clinical follow up, and complications. MAIN OUTCOME MEASUREMENTS: Technical success was decompression of the duct of interest. Clinical success was resolution of jaundice or a > or = 50% reduction in pain or narcotics, as applicable. RESULTS: Between February 2003 and June 2007, EUS-CP was attempted in 20 patients (11 men, 9 women; mean [SD] age 58 +/- 14.9 years). Indications included jaundice (n = 8), biliary stones (n = 3), chronic pancreatitis (n = 6), acute pancreatitis (n = 2), and papillary stenosis (n = 1). Reasons for failed ERCP included periampullary mass (n = 8), intradiverticular papillae (n = 4), and pancreatic duct (PD) stricture (n = 7) or stone (n = 1). Technical success was achieved in 18 of 20 patients (90%). Biliary decompression was obtained in 11 of 12 patients (92%) (7 transpapillary and 4 transenteric-transcholedochal). Pancreatic decompression was obtained in 7 of 8 patients (88%) (3 transpapillary, 4 transgastric). On follow up, clinical improvement was noted in 15 of 20 patients (70%). For treatment of pain associated with chronic pancreatitis, pain scores decreased by a mean of 1.75 (P = .18). Complications (in 2 of 20 [10%]) included perforation (n = 1) and respiratory failure (n = 1). LIMITATIONS: A single-center nonrandomized observational study with a small patient population. CONCLUSIONS: At our academic referral center, single-operator EUS-CP provided decompression of obstructed ducts and may be performed after a failed attempt at conventional ERCP during the same endoscopic session. PMID- 19560770 TI - Protection of sexual and reproductive health rights: addressing violence against women. AB - Violence against women is recognized as a global public health and human rights problem in need of urgent attention. It affects women's health, including their sexual and reproductive health, and their human rights. While progress has been made in the last 15 years, there is still a long way to go. International human rights law and public health provide tools to governments and non-governmental actors to ensure women a life free from violence and its consequences. Health policies and services need to address violence more systematically and health providers must take action. At a minimum, they should be informed and able to respond appropriately to violence, providing appropriate care and referral to other services. Equally, if not more important, is to provide support to interventions that prevent violence against women from happening in the first place. PMID- 19560771 TI - Sensitivity and specificity of a self-administered questionnaire of tobacco use; including the Fagerstrom test. AB - BACKGROUND: Preventing tobacco consumption and promoting cessation among health professionals are of great significance as their habits can influence their patients' attitudes. Knowledge of the prevalence and characteristics of tobacco use in a specific population is important for the design of efficient strategies for preventing people from acquiring the habit and persuading them to stop. Self administered questionnaires are a very common method for determining tobacco use, but assessment is needed of their validity for specific groups, such as occasional smokers or students of health sciences. OBJECTIVES: The aim of this study was to determine the sensitivity and specificity of a self-administered questionnaire for determining tobacco use among a population of young female students of health sciences. As a gold standard, we used a measure of the concentration of cotinine in saliva with different cut-off points. We also analysed the influence on sensitivity and specificity of the questionnaire of nicotine dependence and exposure to secondary smoke. DESIGN: This is an internal validity study (sensitivity and specificity) of a self-administered questionnaire. SETTINGS: The study was carried out in the School of Health Sciences of the University of Leon, at its centres in Leon and Ponferrada. PARTICIPANTS: Data were collected on 432 of a total of 480 women aged under 25 who were studying Health Sciences at the University of Leon in 2007. METHODS: The self-administered questionnaire included data concerning demography, education, tobacco habits and exposure to environmental smoke. Saliva cotinine concentration was determined with an EIA kit. RESULTS: The questionnaire used showed high values of sensitivity and specificity (85.3% and 95.3%) and a very good correlation (Kappa=81.0%) with cotinine values of 10ng/ml or higher. Discrepancies in the test results for those who declared themselves to be smokers only occurred in women with a low nicotine dependency. Among those who did not declare themselves smokers, those exposed to second-hand smoke were 7 times more frequently classified erroneously as smokers than those who had not been exposed. CONCLUSION: The self-administered questionnaire used showed a very good internal validity and a good correlation with cotinine levels of 10ng/mg, and therefore seems to be a good instrument for measuring tobacco use in this population. The availability of information on passive smoking and nicotine dependence is essential for the correct interpretation of the discrepancies. PMID- 19560772 TI - Endothelial activation, inflammation and premature atherosclerosis in children with familial dyslipidemia. AB - OBJECTIVES: Prospective studies demonstrated an increased cardiovascular risk in subjects with high levels of either the endothelial-platelet activation marker P selectin or high-sensitivity C-reactive protein (hs-CRP). Both children with heterozygous familial hypercholesterolemia (FH) and those with familial combined hyperlipidemia (FCHL) are prone to premature atherosclerosis. Our objective was to investigate in children with either FH or FCHL whether P-selectin and hs-CRP contribute to carotid intima-media thickness (IMT), along with increased plasma lipid levels. METHODS: Carotid IMT, serum lipids and soluble P-selectin and hs CRP levels were measured in 88 children (mean age 10.5+/-4.3 years) including 44 dyslipidemic children (25 with FH and 19 with FCHL) and 44 non-dyslipidemic controls. RESULTS: Carotid IMT was significantly higher among dyslipidemic than in control children (0.46+/-0.06mm vs 0.43+/-0.06mm, p=0.003) and serum P selectin levels as well [129(50-254)ng/mL vs 50(24.5-130)ng/mL, p<0.001]. FH but not FCHL children had higher hs-CRP levels than controls [0.7(0.01-6.9)mg/L vs 0.3(0.1-1.2)mg/L, p=0.006]. In the entire sample of dyslipidemic children, carotid IMT was positively associated with soluble P-selectin levels (rho=0.30, p=0.049), but not with hs-CRP. The association between P-selectin and carotid IMT was independent from confounders, including plasma lipid levels. CONCLUSION: Endothelial-platelet activation, more than low-grade systemic inflammation, correlates with premature atherosclerosis among children with familial dyslipidemia, this association being independent from plasma lipid levels. PMID- 19560773 TI - Uncertainty assessment of polychlorinated dibenzo-p-dioxins and dibenzofuran and dioxin-like polychlorinated biphenyl analysis in stationary source sample emissions in accordance with the impending European standard EN-1948 using fly ashes. AB - The analysis of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) present in stack gas emissions and solid residues from incinerators will be mandatory in the foreseeable future. European standard EN-1948 is in the process of being updated through the addition of a new Part 4 related to the analysis of the 12 dl-PCBs. Therefore, either a comprehensive and reliable method capable of analyzing all of these 29 compounds (12 dl-PCBs and 17 2,3,7,8-PCDD/Fs) needs to be developed, or the existing PCDD/F analytical procedure must be adapted to include the dl-PCBs. This study has taken the latter approach of modifying PCDD/F methodology and in particular the fractionation step, by isolating dioxins and dl-PCBs into separate fractions ready for high resolution gas chromatography coupled to high resolution mass spectrometry (HRGC/HRMS) analysis. Results obtained from the analysis of Certified Reference Materials (CRM-490 and CRM-615) and fly ashes from the European Committee for Standardization (CEN) intercalibration study demonstrated that the proposed methodology is appropriate to determine the dl-PCBs in accordance with the impending European standard EN-1948. Uncertainty values obtained during the validation of the analytical methodology were 13% total I-TEQ (International Toxic Equivalent) for PCDD/Fs and 31% total WHO-TEQ (World Health Organization Toxic Equivalent) in the case of dl-PCBs. In addition, 'real' samples such as emissions and fly ashes were successfully analyzed following the proposed analytical method. PMID- 19560774 TI - Effects of extra-column band spreading, liquid chromatography system operating pressure, and column temperature on the performance of sub-2-microm porous particles. AB - The effects of extra-column band spreading, LC system operating pressure, and separation temperature were investigated for sub-2-microm particle columns using both a conventional HPLC system as well as a UPLC system. The contributions of both volume- and time-based extra-column effects were analyzed in detail. In addition, the performance difference between columns containing 2.5 and 1.7 microm particles (same stationary phase) was studied. The performance of these columns was compared using a conventional HPLC system and a low dead volume UPLC system capable of routine operation up to 1000 bar. The system contribution to band spreading and the pressure limitations of the conventional HPLC system were found to be the main difficulties that prevented acceptable performance of the sub-2-microm particle columns. Finally, an increase in operating temperature needs to be accompanied by an increase in flow rate to prevent a loss of separation performance. Thus, at a fixed column length, an increase in temperature is not a substitute for the need for very high operating pressures. PMID- 19560775 TI - Rapid analysis of melamine in infant formula by sweeping-micellar electrokinetic chromatography. AB - In the present study, an analytical method using capillary electrophoresis with on-line preconcentration technique was developed for rapid determination of melamine in infant formula. Both stacking and sweeping preconcentration techniques had been investigated for the comparison of their effectiveness in melamine analysis. The limit of detection of melamine standard was 0.5 ng/mL for the field amplified sample stacking (FASS) technique and 9.2 ng/mL for the sweeping technique. Although the FASS technique provided better concentration efficacy than the sweeping technique, the matrix effect was more profound with the former. Matrix effect was evaluated by comparing the enhancement factor (EF) of melamine standard and post-extraction spiked infant formula solution. The EF was changed from 429.86 +/- 9.81 to the level less than 133.31 with significant peak distortion in the FASS system, and it was remained unchanged in the sweeping system. Sweeping-micellar electrokinetic chromatography (sweeping-MEKC) was demonstrated to be most suitable for real sample analysis. Under optimum sweeping MEKC conditions, melamine content in infant formulas could be determined within 6 min. The developed solid phase extraction (SPE) procedures coupled with the sweeping-MEKC method was subjected to method validation. Run-to-run repeatability (n = 3) and day-to-day reproducibility (n = 3) of peak area were within 3.6% and 4.8% RSD, respectively. The accuracy was tested by spiking 0.5 and 2 microg/mL of melamine standard in the melamine contaminated milk powder provided by the European Commission, and the recoveries were 93.4 +/- 0.5% and 98.7 +/- 0.4%, respectively. Results of this study show a great potential for the sweeping-MEKC method as a tool for the fast screening of melamine in infant formulas. PMID- 19560776 TI - Determination of multi-class pesticides in wines by solid-phase extraction and liquid chromatography-tandem mass spectrometry. AB - This work reports a new sensitive multi-residue liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for detection, confirmation and quantification of forty-six pesticides and transformation products belonging to different chemical classes in wines. The proposed method makes use of a solid-phase extraction (SPE) procedure with Oasis HLB cartridges that combines isolation of the pesticides and sample clean-up in a single step. Analysis is performed by liquid chromatography electrospray ionisation-tandem mass spectrometry (LC-MS/MS) operated in the selected reaction monitoring (SRM) mode, acquiring two specific precursor-product ion transitions per target compound. An investigation of matrix effects has been performed during method validation showing medium to low effects for the majority of the compounds. Limits of detection (LODs) were in the range 0.0003-0.003 mg L( 1) and limits of quantification (LOQs) were in the range 0.001-0.01 mg L(-1). The average recoveries, measured at two concentration levels (0.010 and 0.050 mg L( 1)), were in the range 70-110% for most of the compounds tested with % relative standard deviations below 20%, while a value of 0.010 mg L(-1) has been established as the method limit of quantification (MLOQ) for all target species. Expanded uncertainty values were in the range 10-40% while the Horrat ratios were below 1. The method has been successfully applied to the analysis of 60 wine samples in the course of an annual monitoring study with carbendazim-benomyl, thiophanate-methyl and carbaryl being the most frequently determined pesticides. PMID- 19560777 TI - Quantitative analysis of monoclonal antibodies by cation-exchange chromatofocusing. AB - A robust cation-exchange chromatofocusing method was developed for the routine analysis of a recombinant humanized monoclonal IgG antibody. We compare the chromatofocusing method to the conventional cation-exchange chromatography (CEX) employing a salt gradient and show clear advantages of chromatofocusing over CEX. We demonstrate the suitability of the present chromatofocusing method for its intended purpose by testing the validation characteristics. To our knowledge, this is the first chromatofocusing method developed for the routine analysis of monoclonal antibody charge species. PMID- 19560778 TI - Ionic liquids as superior solvents for headspace gas chromatography of residual solvents with very low vapor pressure, relevant for pharmaceutical final dosage forms. AB - 1-n-Butyl-3-methylimidazolium dimethyl phosphate (BMIM DMP) was identified as the most suitable ionic liquid as solvent for the headspace gas chromatographic analysis of solvents with very low vapor pressure such as dimethylsulfoxide, N methylpyrrolidone, sulfolane, tetralin, and ethylene glycol in a realistic matrix of commonly used excipients (carboxymethylcellulose, magnesium stearate, guar flour, and corn starch) in pharmaceutical products. Limits of quantification and limits of detection were in the low microgram per gram range. The detection of traces of sulfolane in a real sample of tablets containing the drug cefpodoxim proxetil demonstrated the applicability of the method. PMID- 19560779 TI - Mono-distributed single-walled carbon nanotube channel in field effect transistors (FETs) using electrostatic atomization deposition. AB - This communication reports on the novel work of creating a transistor channel based on functionalized single-walled carbon nanotubes (SWNTs) via electrostatic atomization deposition. The current method of drop-cast though convenient was unable to produce replicable transistor device due to its inherent inability in controlling the volume of liquid being drop-cast. Hence, this method of electrostatic atomization was introduced to consistently obtain a uniformly distributed SWNT channel resulting in a good transistor device. PMID- 19560780 TI - Ferrihydrite reactivity toward carbon dioxide. AB - The reaction of ferrihydrite with gaseous CO(2) was investigated with attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and density functional theory (DFT) calculations. ATR-FTIR results show that CO(2) reacts with ferrihydrite resulting in surface adsorbed carbonate species. The carbonate species experimentally observed in view of theoretical calculations are shown to be in large part monodentate binuclear complexes. These carbonate complexes exist as both inner-sphere and outer-sphere hydrogen-bonded complexes. Under "dry" conditions CO(2) reacts with free OH sites on the ferrihydrite surface resulting in a metastable bent CO(2) (bicarbonate-like) complex. Removal of the gaseous reactant leads to the loss of this metastable surface complex. The reaction of CO(2) with hydrated ferrihydrite results in only carbonate formation (no bicarbonate). In this circumstance, experiments and theoretical calculations suggest that hydrogen bound water on surface OH sites prevents the formation of the metastable bicarbonate species. Ferrihydrite that was allowed to react with atmospheric levels of CO(2) and water vapor resulted in the formation of surface carbonate coordinated as both inner and outer-sphere complexes. PMID- 19560782 TI - Location representation in enclosed spaces: what types of information afford young children an advantage? AB - It has been suggested that young children can only reorient, locating a target object, when the geometry of an enclosed space provides distinctive shape information [e.g., Hermer, L., & Spelke, E. (1994). A geometric process for spatial reorientation in young children. Nature, 370, 57-59]. Recently, however, young children were shown to specify location in a square-shaped space, where geometry is uninformative, so long as scale-like information was available on the walls of the space [Huttenlocher, J., & Lourenco, S. F. (2007a). Coding location in enclosed spaces: Is geometry the principle? Developmental Science, 10, 741 746]. Here we build on this work by examining more closely what types of cues afford 18- to 24-month-olds an advantage in locating a target object following disorientation. Their performance was assessed when linear scale-like information was presented either in isolation or in composite form. It was found that, even in isolation, young children searched at the appropriate locations, with added benefit when presented as a composite. We suggest that linear scale-like dimensions, especially when available in composite form, play a critical role in supporting location representation in young children. PMID- 19560781 TI - Immunohistochemical expression of vascular endothelial growth factor and vascular endothelial growth factor receptor in canine cutaneous fibrosarcomas. AB - The expression of five markers associated with tumour angiogenesis, proliferation and apoptosis was studied in 24 canine cutaneous fibrosarcomas. Tumours were assigned histological grades and were immunohistochemically evaluated for the expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2). Additionally, intra-tumour microvessel density (iMVD) was assessed by immunohistochemical labelling for expression of von Willebrand factor (vWf) and tumour proliferation index (PI) was measured following labelling of Ki-67 antigen. Finally, tumour apoptotic index (AI) was determined by application of the terminal deoxynucleotidyl transferase (TdT) mediated dUTP end-labelling method (TUNEL). VEGF and VEGFR-2 expression were detected in 22/24 (92%) and 24/24 (100%) of fibrosarcomas, respectively. There was correlation between VEGF and VEGFR-2 expression (r = 0.51) and between histological grade and PI (r = 0.82). A significant difference in PI between tumours of different histological grade was found (P < 0.05). The median PI in grade 2 and 3 tumours (30.6 and 54.7, respectively) was significantly higher than in grade 1 tumours (6.4). Therefore, only PI correlates significantly with the histological grade of canine cutaneous fibrosarcomas. The potential for autocrine activity for VEGF exists in canine cutaneous fibrosarcomas, as VEGF and VEGFR-2 expression was found in most tumours. PMID- 19560783 TI - Toddlers' referential understanding of pictures. AB - Pictures are referential in that they can represent objects in the real world. Here we explore the emergence of understanding of the referential potential of pictures during the second year of life. In Study 1, 15-, 18-, and 24-month-olds learned a word for a picture of a novel object (e.g., "blicket") in the context of a picture book interaction. Later they were presented with the picture of a blicket along with the real object it depicted and asked to indicate the blicket. Many of the 24-, 18-, and even 15-month-olds indicated the real object as an instance of a blicket, consistent with an understanding of the referential relation between pictures and objects. In Study 2, children were tested with an exemplar object that differed in color from the depicted object to determine whether they would extend the label they had learned for the depicted object to a slightly different category member. The 15-, 18-, and 24-month-old participants failed to make a consistent referential response. The results are discussed in terms of whether pictorial understanding at this age is associative or symbolic. PMID- 19560784 TI - Treatment of neonatal hemochromatosis with exchange transfusion and intravenous immunoglobulin. AB - OBJECTIVE: To determine if immunomodulatory treatment including intravenous immunoglobulin (IVIG) can favorably affect survival in neontatal hemochromatosis (NH) diagnosed postnatally because it can effectively prevent occurrence of NH when applied during gestations at risk. STUDY DESIGN: We treated 16 newborn infants with liver failure due to NH with high-dose IVIG, in combination with exchange transfusion in 13 (ET/IVIG), and compared the outcome with 131 historical controls treated conventionally. RESULTS: The severity of liver disease as estimated by prothrombin time was similar in the subjects receiving ET/IVIG and the historical controls, and the medical therapy was equivalent with the exception of the ET/IVIG therapy. Twelve subjects (75%) had good outcome, defined as survival without liver transplantation, whereas good outcome was achieved in only 17% (23/131) of historical control patients (P < .001). Four subjects died, 2 without and 2 after liver transplant. Survivors were discharged 6 to 90 days after receiving ET/IVIG therapy, and those followed for more than 1 year are within normal measures for growth, development, and liver function. CONCLUSIONS: Immune therapy with ET/IVIG appears to improve the outcome and reduce the need for liver transplantation in patients with NH. PMID- 19560785 TI - IGF-1 mediates PTEN suppression and enhances cell invasion and proliferation via activation of the IGF-1/PI3K/Akt signaling pathway in pancreatic cancer cells. AB - BACKGROUND: Type-1 insulin-like growth factor (IGF-1) up-regulates cell proliferation and invasiveness through activation of PI3K/Akt signaling pathway. IGF-1 also down-regulates the tumor suppressor chromosome 10 (PTEN). We investigated the mechanism by which IGF-1 affects cell proliferation and invasion by suppression of PTEN phosphorylation and interaction with PI3K/PTEN/Akt/NF small ka, CyrillicB signaling pathway in pancreatic cancer. MATERIALS AND METHODS: The expression of IGF-1 receptor (IGF-1R) and PTEN in five pancreatic cancer cell lines was determined by RT-PCR and Western blot. Proliferation and invasion were investigated by WST-1 assay and Matrigel-double chamber assay. Pancreatic cancer cells were transfected with PTEN siRNA to investigate which signaling pathway correlates in regulation of cancer cell proliferation and invasion. RESULTS: Five pancreatic cancer cell lines expressed PTEN and IGF-1R in mRNA and protein levels. Suppression of PTEN phosphorylation strongly enhanced cell proliferation and invasion stimulated with IGF-1 via activation of PI3K/Akt/NF-small ka, CyrillicB signaling pathway. In addition, knockdown of PTEN by siRNA transfection also enhanced activation of PI3K/Akt/NF-small ka, CyrillicB pathway, subsequently up-regulating cell invasiveness and proliferation. CONCLUSIONS: The IGF-1/PI3K/PTEN/Akt/NF-small ka, CyrillicB cascade may be a key pathway stimulating metastasis of pancreatic cancer cells. We suggest that interfering with the functions of IGF-1/PI3K/Akt/NF-small ka, CyrillicB might be a novel therapeutic approach to inhibit aggressive spread of pancreatic cancer. PMID- 19560786 TI - Regional antibiotic delivery for the treatment of experimental prosthetic graft infections. AB - OBJECTIVES: To evaluate the efficacy of antibiotic-impregnated polymethylmethacrylate (PMMA) beads in eradication of an arterial prosthetic graft methicillin-resistant Staphylococcus aureus (MRSA) biofilm in an experimental animal model. METHODS: Forty rats underwent subcutaneous implantation of a MRSA-colonized arterial polytetrafluoroethylene (PTFE) 1 x 1 cm wafer on the back. The effect of regional antibiosis produced by antibiotic PMMA bead placement adjacent to the infected PTFE wafer was determined using four 10 animal study groups: control (no antibiotic), PMMA bead with no antibiotic, PMMA bead with 10% vancomycin, and PMMA bead with 10% daptomycin. After 3 d, the PTFE wafers were explanted and quantitative biofilm cultures, expressed as colony forming units (CFU) per graft wafer, performed using real-time polymerase chain reaction to assess MRSA eradication. No systemic antibiotic was administered. Bioassays of antibiotic bead bacteriocidal were performed by measuring zone of inhibition diameters on MRSA colonized agar culture plates prior to and following graft explantation. RESULTS: All animal tolerated implantation of the MRSA infected PTFE wafer and survived the 3 d until graft explantation. Quantitative biofilm cultures demonstrated a significant decrease (P < 0.01) in MRSA CFUs present on the PTFE wafer surfaces in the presence of both the vancomycin- and daptomycin-impregnated beads compared to controls and plain PMMA beads. Both vancomycin and daptomycin PMMA beads retained antibacterial activity after 3 d of implantation with decrease in zones of inhibition of 15% and 45%, respectively. CONCLUSIONS: Regional antibiotic delivery using an antibiotic-impregnated PMMA bead reduced the bacterial biofilm concentration in experimental subcutaneous pocket model of vascular surgical site infection. The delivery of antibiotics via a PMMA bead may be a useful adjunct in the treatment of vascular surgical site infection. PMID- 19560787 TI - Letter to the editor on a paper by Kimura A, Yoshiro H, Yuasa T. Chronic inflammatory demyelinating polyneuropathy in a patient with hyperIgEemia. J Neurol Sci 2005;231:89-93. PMID- 19560789 TI - Environmental records of anthropogenic impacts on coastal ecosystems: an introduction. PMID- 19560788 TI - Reversible metronidazole-induced cerebellar toxicity in a multiple transplant recipient. AB - Metronidazole-induced central nervous system (CNS) toxicity causes a spectrum of neurological symptoms including ataxia, encephalopathy and peripheral neuropathy. It is associated with characteristic MRI changes of high signal intensity in the dentate nuclei. Given the increasing use of metronidazole, it is import to recognise this drug as a cause of ataxia, as it is entirely reversible on drug withdrawal. PMID- 19560790 TI - [Advances in nonalcoholic fatty liver disease]. PMID- 19560791 TI - Genetic variation and drought response in two Populus x euramericana genotypes through 2-DE proteomic analysis of leaves from field and glasshouse cultivated plants. AB - Genotype and water deficit effects on leaf 2-DE protein profiles of two Populus deltoides x Populus nigra, cv. 'Agathe_F' and 'Cima', were analysed over a short term period of 18 days in glasshouse using 4-month-old rooted cuttings and over a long-lasting period of 86 days in open field using 4-year-old rooted cuttings. Leaf proteomes were analyzed using two-dimensional gel electrophoresis, and proteins were identified after database searching from MS peptide spectra. A reliable genotype effect was observed in the leaf proteome over experiment locations, water regimes and sampling dates. Quantitative differences between genotypes were found. Most of them corresponded to proteins matching isoforms or post-translational modification variants. However, 'Cima' displayed the highest abundance of antioxidant enzymes. In response to water deficit, about 10% of the reproducible spots significantly varied regardless of the experiment location, among which about 25% also displayed genotype-dependent variations. As a whole, while 'Cima' differed from 'Agathe_F' by increased abundance of enzymes involved in photorespiration and in oxidative stress, 'Agathe_F' was mainly differentiated by increased abundance of enzymes involved in photosynthesis. PMID- 19560792 TI - The biosynthetic pathway of crucifer phytoalexins and phytoanticipins: de novo incorporation of deuterated tryptophans and quasi-natural compounds. AB - Although several biosynthetic intermediates in pathways to cruciferous phytoalexins and phytoanticipins are common, questions regarding the introduction of substituents at N-1 of the indole moiety remain unanswered. Toward this end, we investigated the potential incorporations of several perdeuterated d- and l-1' methoxytryptophans, d- and l-tryptophans and other indol-3-yl derivatives into pertinent phytoalexins and phytoanticipins (indolyl glucosinolates) produced in rutabaga (Brassica napus L. ssp. rapifera) roots. In addition, we probed the potential transformations of quasi-natural compounds, these being analogues of biosynthetic intermediates that might lead to "quasi-natural" products (products similar to natural products but not produced under natural conditions). No detectable incorporations of deuterium labeled 1'-methoxytryptophans into phytoalexins or glucobrassicin were detected. l-tryptophan was incorporated in a higher percentage than d-tryptophan into both phytoalexins and phytoanticipins. However, in the case of the phytoalexin rapalexin A, both d- and l-tryptophan were incorporated to the same extent. Furthermore, the transformations of both 1' methylindolyl-3'-acetaldoxime and 1'-methylindolyl-3'-acetothiohydroxamic acid (quasi-natural products) into 1'-methylglucobrassicin but not into phytoalexins suggested that post-aldoxime enzymes in the biosynthetic pathway of indolyl glucosinolates are not substrate-specific. Hence, it would appear that the 1 methoxy substituent of the indole moiety is introduced downstream from tryptophan and that the post-aldoxime enzymes of the glucosinolate pathway are different from the enzymes of the phytoalexin pathway. A higher substrate specificity of some enzymes of the phytoalexin pathway might explain the relatively lower structural diversity among phytoalexins than among glucosinolates. PMID- 19560794 TI - A 2D strain estimator with numerical optimization method for soft-tissue elastography. AB - Elastography is a bioelasticity-based imaging modality which has been proved to be a potential evaluation tool to detect the tissue abnormalities. Conventional method for elastography is to estimate the displacement based on cross correlation technique firstly, then strain profile is calculated as the gradient of the displacement. The main problem of this method arises from the fact that the cross-correlation between pre- and post-compression signals will be decreased because of the signal's compression-to-deformation. It may constrain the estimation of the displacement. Numerical optimization, as an efficient tool to estimate the non-rigid deformation in image registration, has its potential to achieve the elastogram. This paper incorporates the idea of image registration into elastography and proposes a radio frequency (RF) signal registration strain estimator based on the minimization of a cost function using numerical optimization method with Powell algorithm (NOMPA). To evaluate the proposed scheme, the simulation data with a hard inclusion embedded in the homogeneous background is produced for analysis. NOMPA can obtain the displacement profiles and strain profiles simultaneously. When compared with the cross-correlation based method, NOMPA presents better signal-to-noise ratio (SNR, 32.6+/-1.5 dB vs. 23.8+/-1.1 dB) and contrast-to-noise ratio (CNR, 28.8+/-1.8 dB vs. 21.7+/-0.9 dB) in axial normal strain estimation. The in vitro experiment of porcine liver with ethanol-induced lesion is also studied. The statistic results of SNR and CNR indicate that strain profiles by NOMPA performs better anti-noise and target detectability than that by cross-correlation based method. Though NOMPA carry a heavier computational burden than cross-correlation based method, it may be an useful method to obtain 2D strains in elastography. PMID- 19560793 TI - Laser acupuncture for acute inflammatory, visceral and neuropathic pain relief: An experimental study in the laboratory rat. AB - Laser acupuncture is defined as the stimulation of traditional acupuncture points with low-intensity, non-thermal laser irradiation. We explored the clinical efficacy of a very low level diode laser wavelength 670 nm (Biolite LP020, RGM, Genoa, Italy), used to stimulate acupoints ST36 Zu San Li and TH5 Waiguan, on well-established experimental models of acute and persistent pain in the rat, e.g. acute inflammatory pain, muscle pain, visceral pain and neuropathic pain. We report the anti-edema and anti-hyperalgesia effects of laser acupuncture in models of acute inflammatory pain, e.g. CFA-induced inflammation and myofascial pain. We also indicate that spontaneous pain and thermal hyperalgesia are reduced in a neuropathic pain model, e.g. axotomy. On the contrary, no effects due to laser-acupuncture were observed on discomfort indices in a model of visceral pain, e.g. cystitis due to cyclophosphamide. We thus provide evidences that acupoints stimulation using a very low intensity laser irradiation can control pain and edema in specific experimental conditions. PMID- 19560796 TI - Remediation of trichloroethylene by bio-precipitated and encapsulated palladium nanoparticles in a fixed bed reactor. AB - Trichloroethylene is a toxic and recalcitrant groundwater pollutant. Palladium nanoparticles bio-precipitated on Shewanella oneidensis were encapsulated in polyurethane, polyacrylamide, alginate, silica or coated on zeolites. The reactivity of these bio-Pd beads and zeolites was tested in batch experiments and trichloroethylene dechlorination followed first order reaction kinetics. The calculated k-values of the encapsulated catalysts were a factor of six lower compared to non-encapsulated bio-Pd. Bio-Pd, used as a catalyst, was able to dechlorinate 100 mgL(-1) trichloroethylene within a time period of 1h. The main reaction product was ethane; yet small levels of chlorinated intermediates were detected. Subsequently polyurethane cubes empowered with bio-Pd were implemented in a fixed bed reactor for the treatment of water containing trichloroethylene. The influent recycle configuration resulted in a cumulative removal of 98% after 22 h. The same reactor in a flow through configuration achieved removal rates up to 1059 mg trichloroethylene g Pd(-1)d(-1). This work showed that fixed bed reactors with bio-Pd polyurethane cubes can be instrumental for remediation of water contaminated with trichloroethylene. PMID- 19560795 TI - Pilot-scale electrokinetic movement of HCB and Zn in real contaminated sediments enhanced with hydroxypropyl-beta-cyclodextrin. AB - This study deals with the efficiency of a pilot-scale electrokinetic (EK) treatment on real aged sediments contaminated with hexachlorobenzene (HCB) and Zn. A total of 0.5m(3) of sediments were treated under a constant voltage in a polyvinyl chloride reactor. The changes of sediment pH, electrical conductivity (EC), organic content (OC), the transport of contaminants in sediments and the consumption of electric energy were evaluated. After 100 d processing, sediment pH slightly increased compared with the initial values, particularly in the bottom layer close to cathodic section, while sediment EC in most sections significantly decreased. Sediment OC in all sections increased, which implied that hydroxypropyl-beta-cyclodextrin (HPCD) was successfully penetrated across sediments by electroosmosis. Significant movement of contaminants was observed across sediments with negligible removals. Both HCB and Zn generally moved from sections near anode and accumulated near cathode. Upon the completion of treatment, the electric energy consumption was calculated as 563 kWhm(-3). This pilot-scale EK test indicates that it is difficult to achieve great removal of hydrophobic organic compounds (HOCs), or HOCs and heavy metal mixed contaminants, by EK treatment in large scale with the use of HPCD. PMID- 19560798 TI - Hexachlorocyclohexanes (HCH) in ringed seal (Phoca hispida) from Ulukhaktok (Holman), NT: trends from 1978 to 2006. AB - Trends in alpha-, beta-, and gamma-hexachlorocyclohexane (HCH) concentrations were examined in blubber lipid of ringed seals (Phoca hispida) from Ulukhaktok (Holman), NT (Canada) sampled at intervals between 1978 and 2006. alpha-HCH usually represented approximately 90% of the total HCH isomers. alpha-HCH and gamma-HCH concentrations showed no change over the sampling interval, but beta HCH concentrations increased significantly, about 8-10-fold in females and 4-5 fold in males. Residue concentrations showed no dependence on age. Concentrations (all data as ng/g lipid, GM (range)) of alpha-HCH were significantly higher (P<0.001 by t-test) in males (217 (93.9-517), n=37) than those in females (138 (40.9-402), n=38). beta-HCH concentrations did not differ between the sexes. Concentrations of gamma-HCH were significantly higher (P<0.05) in males (6.74 (0 46.7)) than in females (4.35 (0-19.0)). Although global emissions of both alpha HCH and beta-HCH have declined since the early 1980's, the "signal" of HCH emission changes has not yet resulted in a "response" in ringed seal residue concentrations. In the light of our current understanding of the dynamics of HCH in the Arctic, we conclude that any such response may not be detected by retrospective analyses of the sort describe here at least for another decade or so, because of the longevity of the seals. PMID- 19560797 TI - Method for the determination of selected organochlorine pesticides and polychlorinated biphenyls in human serum based on a gel permeation chromatographic clean-up. AB - A gel permeation chromatographic (GPC) clean-up based method was developed for determination of selected organochlorine pesticides and polychlorinated biphenyls in human serum. The method permits automation of the sample extract clean-up stage and is designed to work with small amounts of sample. Different working variables were studied in its development, including injection volume, flow rate, and fat amount as the most representative coextract. The method provides solvent and time savings. Initial extraction was performed using 96-well solid-phase disk extraction plates, and quantification was performed by gas chromatography with electron capture detection and mass spectrometry. Recoveries of PCB congeners 28, 52, 101, 118, 138, 153, and 180 and organochlorine pesticides HCB, beta-HCH, gamma-HCH, heptachlor epoxide, p,p'-DDE, and p,p'-DDT at two spiking levels ranged from 55% to 115% with relative standard deviations ranging from 2.0% to 14.6%. Limits of quantification ranged from 0.06 to 0.16 ng mL(-1) for PCBs and from 0.12 to 0.36 ng mL(-1) for organochlorine pesticides. Finally, the method was applied to the analysis of 38 serum samples and the results were compared with those of another procedure validated at the laboratory. PMID- 19560799 TI - Topical application of dressing with amino acids improves cutaneous wound healing in aged rats. AB - The principal goal in treating surgical and non-surgical wounds, in particular for aged skin, is the need for rapid closure of the lesion. Cutaneous wound healing processes involve four phases including an inflammatory response with the induction of pro-inflammatory cytokines. If inflammation develops in response to bacterial infection, it can create a problem for wound closure. Reduced inflammation accelerates wound closure with subsequent increased fibroblast function and collagen synthesis. On the contrary, prolonged chronic inflammation results in very limited wound healing. Using histological and immunohistochemical techniques, we investigated the effects of a new wound dressing called Vulnamin that contains four essential amino acids for collagen and elastin synthesis plus sodium ialuronate (Na-Ial), compared with Na-Ial alone, in closure of experimental cutaneous wounds of aged rats. Our results showed that the application of Vulnamin dressings modulated the inflammatory response with a reduction in the number of inflammatory cells and inducible nitric oxide synthase (iNOS) immunolocalisation, while increasing endothelial nitric oxide synthase (eNOS) and transforming growth factor-beta1 (TGF-beta1) immunolocalisation. Furthermore, the dressing increased the distribution density of fibroblasts and aided the synthesis of thin collagen fibers resulting in a reduction in healing time. The nutritive approach using this new wound dressing can provide an efficacious and safe strategy to accelerate wound healing in elderly subjects, simplifying therapeutic procedures and leading to an improved quality of life. PMID- 19560800 TI - Effects of mineral trioxide aggregate on the differentiation of rat dental pulp cells. AB - The effect of mineral trioxide aggregate (MTA) on the odontoblast-like differentiation of pulp cells was evaluated using heat-shock protein 25 (hsp25) as a marker for odontoblast differentiation. The cells were cultured with tooth colored MTA or calcium hydroxide-containing cement (Dycal). The effects of the materials on the pulp cells were observed using a confocal laser scanning microscope. The cells were labelled immunocytochemically using polyclonal antibodies against hsp25 and actin. The mRNA expression of hsp25 and dspp in the pulp cells at 2 days were examined by quantitative reverse transcription polymerase chain reaction (RT-PCR). Most of the cells cultured with MTA showed an intense immunolabelling for hsp25 and the mRNA expressions of hsp25 and dspp at 2 days were higher than those cultured with Dycal. These findings indicate that MTA is an effective pulp capping material and is able to induce the differentiation of odontoblast-like cells and the formation of reparative tertiary dentin with minimum apoptosis. PMID- 19560801 TI - Young patients with endometrial cancer: how many could be eligible for fertility sparing treatment? AB - OBJECTIVES: To assess the characteristics of young women with endometrial carcinoma, and evaluate those potentially eligible for conservative therapy. METHODS: We identified women diagnosed with endometrial cancer between 1970 and 2005 at the population-based Geneva Cancer Registry (n=1365). We classified patients into two age groups (< or =45 and >45 years old). Differences in demographic, tumor, diagnostic and treatment characteristics were tested with chi square. Kaplan-Meier analysis was used to calculate survival from endometrial cancer and the log-rank test to analyze differences in survival between the two groups. RESULTS: The young group comprised 44 (3.2%) women and the old group 1321 (96.8%) women. Synchronous ovarian malignancies were found in six patients (14%) in the young group, compared with 23 (2%) in the old group (P<0.001). Tumor stage was also different between the two groups, principally because of more stage II among the young (P=0.012). Histological tumor type, grade and specific endometrial cancer 5-year survival did not significantly differ between the two groups. According to final histopathologic evaluation, eight patients from the young group had FIGO stage IA, grade I disease, i.e. may have been eligible for fertility-sparing treatment, corresponding to an incidence rate of 0.3/100,000. CONCLUSION: No significant difference regarding tumor characteristics and survival between young and older patients was observed, except stage of disease and rate of synchronous ovarian malignancy. Conservative approach is a meaningful quality of life goal for patients with cancer, but only suitable for a limited number of patients. PMID- 19560802 TI - Food allergy and food allergy attitudes among college students. AB - BACKGROUND: Little information is known about food allergy among college students. OBJECTIVE: We sought to assess food allergy trends and behavioral attitudes on a large university campus. METHODS: An online survey was distributed by e-mail to local university undergraduate students. Symptom severity was determined based on previously published criteria for anaphylaxis. RESULTS: A total of 513 individuals responded, with 57% reporting an allergic reaction to food. Of this group, 36.2% reported symptoms consistent with anaphylaxis, and these reactions frequently occurred while enrolled. Allergy to milk (P = .032), tree nut (P < .0001), shellfish (P < .0001), and peanut (P < .0001) was significantly associated with having symptoms of anaphylaxis. Some form of emergency medication was reportedly maintained in 47.7%, including self injectable epinephrine (SIE; 21%), although only 6.6% reported always carrying this device. Medication maintenance was significantly lower among students who had not had a reaction while enrolled (P < .0001). Only 39.7% reported always avoiding foods to which they were allergic. Within the group that reported intentionally consuming known allergens, there were significantly lower numbers of individuals who reported carrying SIE (P < .0001) and significantly higher numbers of individuals with a history of a reaction that had not resulted in symptoms of anaphylaxis (P = .026). CONCLUSION: Potentially life-threatening anaphylactic reactions to foods are occurring on college campuses. Only 39.7% of students with food allergy avoided a self-identified food allergen, and more than three fourths did not maintain SIE. Such behaviors might place these students at increased risk for adverse events. PMID- 19560803 TI - Summary of the 2008 National Institute of Allergy and Infectious Diseases-US Food and Drug Administration Workshop on Food Allergy Clinical Trial Design. AB - This article summarizes the proceedings of a 2008 Workshop on Food Allergy Clinical Trials Design co-organized by the National Institute of Allergy and Infectious Diseases and the US Food and Drug Administration. The use of food allergens both as therapy and for oral food challenges is associated with a risk of anaphylaxis. Investigators are strongly encouraged to address regulatory considerations by discussing proposed studies with the US Food and Drug Administration. Food allergen administration through the oral or sublingual routes might be less risky than through the subcutaneous route, but this hypothesis has not been proved, and subjects with food allergy might still be at high risk of allergic reactions to such allergen administration. Two distinct mechanisms might lead to beneficial clinical outcomes: desensitization (reversible when food allergen therapy is stopped) and tolerance (persistent benefit even after allergen therapy is stopped). There are important clinical distinctions between desensitization and tolerance. The efficacy of a therapy for food allergy can be evaluated by assessing changes in the dose response to double blind, placebo-controlled oral food challenges before and after therapy and also by assessing changes in the number of allergic episodes during a longitudinal natural history/exposure study; both approaches have strengths and limitations. PMID- 19560804 TI - The safety of initiating Hymenoptera immunotherapy at 1 microg of venom extract. PMID- 19560805 TI - Frozen-thawed rhinoceros sperm exhibit DNA damage shortly after thawing when assessed by the sperm chromatin dispersion assay. AB - This study reports on the successful validation (via in situ nick translation and neutral comet assay) of the equine Sperm-Halomax kit as an appropriate methodology for the assessment of sperm DNA fragmentation in three species of rhinoceros. Rhinoceros sperm nuclei with fragmented DNA (validated using in situ nick translation) were evident as large halos with dispersed DNA fragments, whereas those with nonfragmented DNA displayed small halos of nondispersed DNA within the microgel. There was a high correlation (r) of 0.974 (R(2) value=0.949; P<0.01; n=16) between the respective assessments of the Sperm Chromatin Dispersion test (SCDt) and the neutral comet assay. Application of the SCDt to determine the DNA fragmentation dynamics of rhinoceros (n=6) sperm frozen in liquid nitrogen vapor and incubated postthaw at 37 degrees C for up to 48 h to mimic in vitro conditions in the female reproductive tract, revealed an increase (P=0.001) in DNA damage, as soon as 4h after the start of incubation. Linear regression equations were calculated for all six rhinoceroses over the first 6h of incubation and revealed individual animal variation. Freshly collected and incubated (37 degrees C) rhinoceros (n=3) sperm had no increase in the basal level of DNA fragmentation for up to 48 h, indicating that the cryopreservation of rhinoceros sperm in liquid nitrogen vapor, as used in this study, appeared to result in freeze-thaw DNA damage. PMID- 19560806 TI - Use of random amplified polymorphic DNA to identify several novel markers for sex identification in the crested serpent eagle and crested goshawk. AB - The crested serpent eagle (Spilornis cheela hoya) has no distinct sexual dimorphic traits. In the current study, we report the results of an EE0.6 (EcoRI 0.6-kb fragment) sequence applied to S. cheela hoya and a novel random amplified polymorphic DNA (RAPD) marker that can be used to sex individuals within the species S. cheela hoya and Accipiter trivigatus formosae (crested goshawk). We used sex-specific primers for the avian CHD1 (chromo-helicase-DNA-binding 1) gene and the EE0.6 sequence in PCR assays to determine sex. In addition, 120 random primers were used for RAPD fingerprinting to search for novel sex-specific fragments of S. cheela hoya. The OPBB08 random primer generated a 1241-bp sex specific fragment in all female S. cheela hoya. From the nucleotide sequence, PCR primers were designed to amplify 553-, 895-, and 194-bp sex-specific fragments present in all female S. cheela hoya. One of these primer pairs (ScBB08-7F/R) also amplified a male/female common fragment that can be used as an internal control (543bp). Moreover, one of the primer pairs (ScBB08-5aF/5bR) could be used to identify genders of A. trivigatus formosae. In conclusion, we identified novel sex-specific DNA markers of S. cheela hoya and A. trivigatus formosae that can be used for rapid and accurate sex identification. PMID- 19560807 TI - Deformation analysis of the periodontium considering the viscoelasticity of the periodontal ligament. AB - OBJECTIVES: The aim of the present work was, by means of a combined experimental and numerical approach, to investigate the full-field distributions of displacement, stress and strain, and their evolution with loading in the entire fresh periodontium under an externally applied force. METHODS: In situ intrusion tests were performed to identify the nonlinear, viscoelastic behavior of the periodontal ligament (PDL) of a pig mandible; a digital image correlation method was applied to examine the full-field deformation patterns in the entire periodontium. The finite element (FE) model was created based on the actual anatomic profiles of individual constituents of the tooth structure; the nonlinear and time-dependent viscoelastic properties of the PDL were input into the FE model to fit the numerical computations with the experimental measurements. RESULTS: The nonlinear, viscoelastic behavior of the PDL was identified and characterized quantitatively. The simulation results were validated by the experiments. The results showed the tilting of tooth and the movement of cervical bone toward the mid-tooth in the studied periodontium under vertical compressive loading. Major strain was concentrated in the PDL, with the maxima near to the tooth apexes, at the tooth-root bifurcation and also at the sides of the tooth roots, and maintained a slight rise during holding of the applied displacement. High stress in the tooth was located mainly at the sides of tooth roots, in the bone it was concentrated near the apexes and the root bifurcation, and these stresses decreased gradually during the holding period. SIGNIFICANCE: The combined approach of experiments that apply the digital image correlation method and FE analyses that take into account the nonlinear and time dependent viscoelasticity of the PDL enables the acquisition of a full picture of detailed, realistic stress/strain fields and deformation patterns of the entire fresh periodontium, being of essence in orthodontics and dentistry. PMID- 19560808 TI - Systematic review and meta-analysis of interventions relevant for young offenders with mood disorders, anxiety disorders, or self-harm. AB - Background Mood and anxiety disorders, and problems with self-harm are significant and serious issues that are common in young people in the Criminal Justice System. Aims To examine whether interventions relevant to young offenders with mood or anxiety disorders, or problems with self-harm are effective. Method Systematic review and meta-analysis of data from randomised controlled trials relevant to young offenders experiencing these problems. Results An exhaustive search of the worldwide literature (published and unpublished) yielded 10 studies suitable for inclusion in this review. Meta-analysis of data from three studies (with a total population of 171 individuals) revealed that group-based Cognitive Behaviour Therapy (CBT) may help to reduce symptoms of depression in young offenders. Conclusions These preliminary findings suggest that group-based CBT may be useful for young offenders with such mental health problems, but larger high quality RCTs are now needed to bolster the evidence-base. PMID- 19560809 TI - Treatment of rheumatoid arthritis: we are getting there. PMID- 19560811 TI - Colonisation of fish and crabs of wave energy foundations and the effects of manufactured holes - a field experiment. AB - Several Western European countries are planning for a significant development of offshore renewable energy along the European Atlantic Ocean coast, including many thousands of wave energy devices and wind turbines. There is an increasing interest in articulating the added values of the creation of artificial hard bottom habitats through the construction of offshore renewable energy devices, for the benefit of fisheries management and conservation. The Lysekil Project is a test park for wave power located about 100 km north of Gothenburg at the Swedish west coast. A wave energy device consists of a linear wave power generator attached to a foundation on the seabed, and connected by a wire to a buoy at the surface. Our field experiment examined the function of wave energy foundations as artificial reefs. In addition, potentials for enhancing the abundance of associated fish and crustaceans through manufactured holes of the foundations were also investigated. Assemblages of mobile organisms were examined by visual censuses in July and August 2007, 3 months after deployment of the foundations. Results generally show low densities of mobile organisms, but a significantly higher abundance of fish and crabs on the foundations compared to surrounding soft bottoms. Further, while fish numbers were not influenced by increased habitat complexity (holes), it had a significantly positive effect on quantities of edible crab (Cancer pagurus), on average leading to an almost five fold increase in densities of this species. Densities of spiny starfish (Marthasterias glacialis) were negatively affected by the presence of holes, potentially due to increased predator abundance (e.g. C. pagurus). These results suggest a species-specific response to enhanced habitat complexity. PMID- 19560812 TI - Chemical conjugation of urokinase to magnetic nanoparticles for targeted thrombolysis. AB - Thrombolytic therapy is an important treatment for thrombosis, a life-threatening condition in cardiovascular diseases. However, the traditional thrombolytic therapies have often been associated with the risk of severe bleeding. By conjugating urokinase with magnetic nanoparticles (MNPs), we have presented a strategy to control thrombolysis within a specific site. The covalent bioconjugate of urokinase and dextran-coated MNPs was synthesized and isolated. Thrombolysis by the conjugate was studied under a magnetic field in a rat arteriovenous shunt thrombosis model. The magnetic field was generated by two AlNiCo permanent magnets around the site of thrombus. The magnetic field enhanced the thrombolytic efficacy of the conjugate by 5-fold over urokinase with no reduction in plasma fibrinogen and little prolonged bleeding time. It suggested that thrombolysis had been specifically directed to the desired site by the magnetic carrier under the magnetic field. Additionally, the conjugate had a longer half-life than urokinase in circulation. PMID- 19560810 TI - Golimumab in patients with active rheumatoid arthritis after treatment with tumour necrosis factor alpha inhibitors (GO-AFTER study): a multicentre, randomised, double-blind, placebo-controlled, phase III trial. AB - BACKGROUND: Tumour necrosis factor alpha (TNFalpha) inhibitors are frequently used to treat rheumatoid arthritis, but whether use of a different TNFalpha inhibitor can improve patient response is unknown. We assess the efficacy and safety of the TNFalpha inhibitor golimumab in patients with active rheumatoid arthritis who had previously received one or more TNFalpha inhibitors. METHODS: 461 patients with active rheumatoid arthritis from 82 sites in 10 countries were randomly allocated by interactive voice response system, stratified by study site and methotrexate use, to receive subcutaneous injections of placebo (n=155), 50 mg golimumab (n=153), or 100 mg golimumab (n=153) every 4 weeks between Feb 21, 2006, and Sept 26, 2007. Allocation was double-blind. Eligible patients had been treated with at least one dose of a TNFalpha inhibitor previously. Patients continued stable doses of methotrexate, sulfasalazine, hydroxychloroquine, oral corticosteroids, and non-steroidal anti-inflammatory drugs. The primary endpoint was achievement at week 14 of 20% or higher improvement in American College of Rheumatology criteria for assessment of rheumatoid arthritis (ACR20). At week 16, patients who had less than 20% improvement in tender and swollen joint counts were given rescue therapy and changed treatment from placebo to 50 mg golimumab, or from 50 mg to 100 mg golimumab. Drug efficacy was assessed by intention to treat and safety was assessed according to the study drug given. This study is registered with ClinicalTrials.gov, number NCT00299546. FINDINGS: Patients had discontinued previous TNFalpha inhibitors because of lack of effectiveness (269 [58%] patients) or reasons unrelated to effectiveness (246 [53%] patients), such as intolerance and accessibility issues. Patients had active disease, which was indicated by a median of 14.0 (IQR 9.0-22.0) swollen and 26.0 (16.0-41.0) tender joints for the whole group. 28 (18%) patients on placebo, 54 (35%) patients on 50 mg golimumab (odds ratio 2.5 [95% CI 1.5-4.2], p=0.0006), and 58 (38%) patients on 100 mg golimumab (2.8 [1.6-4.7], p=0.0001) achieved ACR20 at week 14. Two patients were never treated, and 57 patients did not complete the study because of adverse events, unsatisfactory treatment effect, loss to follow-up, death, or other reasons. 155 patients on placebo, 153 on 50 mg golimumab, and 153 on 100 mg golimumab were assessed for drug efficacy. For weeks 1-16, serious adverse events were recorded in 11 (7%) patients on placebo, 8 (5%) on 50 mg golimumab, and 4 (3%) on 100 mg golimumab. For weeks 1-24, after some patients were given rescue therapy, serious adverse events were recorded in 15 (10%) patients on placebo, 14 (5%) on 50 mg golimumab, and 8 (4%) on 100 mg golimumab. INTERPRETATION: Golimumab reduced the signs and symptoms of rheumatoid arthritis in patients with active disease who had previously received one or more TNFalpha inhibitors. FUNDING: Centocor Research and Development and Schering-Plough Research Institute. PMID- 19560813 TI - Functional PEG-peptide hydrogels to modulate local inflammation induced by the pro-inflammatory cytokine TNFalpha. AB - Hydrogels are an important class of biomaterials for cell encapsulation and delivery, providing a physical barrier or "immuno-isolation" between the host tissue and encapsulated cells. The semi-permeable gel protects the encapsulated cells from host immune cells and/or antibody recognition while allowing facile diffusion of nutrients. However, a previously un-addressed problem is that highly permissive hydrogels cannot exclude the infiltration of soluble immune-mediators, such as pro-inflammatory cytokines that are highly expressed in wounded environments in vivo. When encountered with pro-inflammatory cytokines, encapsulated cells fail to perform their desired functions. Here, we report the synthesis, characterization, and application of peptide-functionalized, cytokine antagonizing poly(ethylene glycol) (PEG) hydrogels capable of sequestering the pro-inflammatory cytokine, tumor necrosis factor alpha (TNFalpha). Results demonstrate that the survival, function, and differentiation of encapsulated cells (e.g., rat adrenal pheochromocytoma cells--PC12s, mouse pancreatic islets, and human mesenchymal stem cells or hMSCs) are significantly hindered in un modified PEG hydrogels under in vitro TNFalpha treatments. In contrast, cells encapsulated in TNFalpha-antagonizing hydrogels are un-affected by the infiltrated TNFalpha. This study demonstrates the importance of controlling the availability of pro-inflammatory cytokines in highly permissive hydrogels. PMID- 19560814 TI - In vitro cellular responses to scaffolds containing two microencapulated growth factors. AB - Growth factors play an important role in the complex cascade of tissue events in periodontal regeneration, although optimal methods of delivery remain to be identified. We hypothesize that multiple delivery of growth factors, particularly via a microparticle-containing scaffold, will enhance cellular events leading to periodontal regeneration. In this study, cellular responses of periodontal ligament fibroblasts (PDLFs) in scaffolds containing microparticles (MPs) loaded with either bone morphogenetic protein (BMP)-2, insulin-like growth factor (IGF) 1, or a mixture of both MPs were evaluated, and the dual-MP-containing scaffold exhibited the release of different proteins in a sustained and independent fashion. When PDLF-seeded scaffolds were cultured in a flow perfusion bioreactor, cell metabolism and proliferation of PDLFs were significantly increased within 3 days in all IGF-1-containing scaffolds compared with those in groups lacking IGF 1 and particulate delivery enhanced these effects between 3 and 7 days. The dual MP-containing group showed the most positive results. Both the BMP-2-in-MP and IGF-1-in-MP groups showed greater effects of alkaline phosphatase activity, more osteocalcin and osteopontin production, and more calcium deposition compared with matched GF-adsorbed groups. All osteoblastic markers were at their highest in the dual-MP-containing group at all detected time points. The combined results suggest that our dual-MP-containing scaffold can be used as a cell vehicle to positively affect cell behavior, thus exhibiting the potential to be a candidate scaffold for future periodontal tissue engineering. PMID- 19560815 TI - In-situ crosslinking hydrogels for combinatorial delivery of chemokines and siRNA DNA carrying microparticles to dendritic cells. AB - Polymer-based, injectable systems that can simultaneously deliver multiple bioactive agents in a controlled manner could significantly enhance the efficacy of next generation therapeutics. For immunotherapies to be effective, both prophylactically or therapeutically, it is not only critical to drive the antigen (Ag)-specific immune response strongly towards either T helper type 1 (Th1) or Th2 phenotype, but also to promote recruitment of a high number of antigen presenting cells (APCs) at the site of immunization. We have recently reported a microparticle-based system capable of simultaneously delivering siRNA and DNA to APCs. Here we present an in-situ crosslinkable, injectable formulation containing dendritic cell (DC)-chemo-attractants and dual-mode DNA-siRNA loaded microparticles to attract immature DCs and simultaneously deliver, to the migrated cells, immunomodulatory siRNA and plasmid DNA antigens. These low crosslink density hydrogels were designed to degrade within 2-7 days in vitro and released chemokines in a sustained manner. Chemokine carrying gels attracted 4-6 folds more DCs over a sustained period in vitro, compared to an equivalent bolus dose. Interestingly, migrated DCs were able to infiltrate the hydrogels and efficiently phagocytose the siRNA-DNA carrying microparticles. Hydrogel embedded microparticles co-delivering Interleukin-10 siRNA and plasmid DNA antigens exhibited efficient Interleukin-10 gene knockdown in migrated primary DCs in vitro. PMID- 19560817 TI - The application of poly (glycerol-sebacate) as biodegradable drug carrier. AB - Poly (glycerol-sebacate) (PGS) is an elastomeric biodegradable polymer which possesses the ideal properties of drug carriers. In the present study, we prepared a series of PGS implants (5-FU-PGSs) loaded with different weight percent of 5-fluorouracil (2, 5, 7.5 and 10%). We studied the infrared spectrum properties, in vitro degradation and drug release, in vivo degradation and tissue biocompatibility of 5-FU-PGSs, in order to provide detailed information for the application of PGS as biodegradable drug carrier in cancer therapy. Macroscopically, all 5-FU-PGS wafers in phosphate buffer solution (PBS) kept their geometries during the degradation period of 30 days. The in vitro degradation rates of 5-FU-PGSs were accelerated when higher concentration of 5-FU was doped. Scanning electron microscopy observation showed that the surfaces of 5 FU-PGSs with higher concentration of 5-FU had irregular pits. The cumulative drug release profiles of 5-FU-PGSs exhibited a biphasic release with an initial burst release in the first day. After 7 days, almost 100% cumulative release of 5-FU was found for all 5-FU-PGSs.The degradation rate of 5-FU-PGSs in vivo was much quicker than that in vitro. Hematoxylin and eosin staining showed that no remarkable inflammations were observed in the tissue surrounding 5-FU-PGS implants, suggesting 5-FU-PGSs had good biocompatibility and no tissue toxicity. In vitro anti-tumor activity assay suggested that 5-FU-PGSs exhibited anti-tumor activity through sustained-release drug mode. These results demonstrate that PGS is a candidate of biodegradable drug carriers. PMID- 19560816 TI - A star-PEG-heparin hydrogel platform to aid cell replacement therapies for neurodegenerative diseases. AB - Biofunctional matrices for in vivo tissue engineering strategies must be modifiable in both biomolecular composition and mechanical characteristics. To address this challenge, we present a modular system of biohybrid hydrogels based on covalently cross-linked heparin and star-shaped poly(ethylene glycols) (star PEG) in which network characteristics can be gradually varied while heparin contents remain constant. Mesh size, swelling and elastic moduli were shown to correlate well with the degree of gel component cross-linking. Additionally, secondary conversion of heparin within the biohybrid gels allowed the covalent attachment of cell adhesion mediating RGD peptides and the non-covalent binding of soluble mitogens such as FGF-2. We applied the biohybrid gels to demonstrate the impact of mechanical and biomolecular cues on primary nerve cells and neural stem cells. The results demonstrate the cell type-specific interplay of synergistic signaling events and the potential of biohybrid materials to selectively stimulate cell fate decisions. These findings suggest important future uses for this material in cell replacement based-therapies for neurodegenerative diseases. PMID- 19560818 TI - Controlled drug release from multilayered phospholipid polymer hydrogel on titanium alloy surface. AB - Here we describe the functionalization of a multilayered hydrogel layer on a Ti alloy with an antineoplastic agent, paclitaxel (PTX). The multilayered hydrogel was synthesized via layer-by-layer self-assembly (LbL) using selective intermolecular reactions between two water-soluble polymers, phospholipid polymer (PMBV) containing a phenylboronic acid unit and poly(vinyl alcohol) (PVA). Reversible covalent bonding between phenylboronic acid and the polyol provided the driving force for self-assembly. Poorly water-soluble PTX dissolves in PMBV aqueous solutions because PMBV is amphiphilic. Therefore, our multilayered hydrogel could be loaded with PTX at different locations to control the release profile and act as a drug reservoir. The amount of PTX incorporated in the hydrogel samples increased with the number of layers but was not directly proportional to the number of layers. However, as the step for making layers was repeated, the concentration of PTX in the PMBV layers increased. The different solubilities of PTX in PMBV and PVA aqueous solutions allow for the production of multilayered hydrogels loaded with PTX at different locations. In vitro experiments demonstrated that the location of PTX in the multilayered hydrogel influences the start and profile of PTX release. We expect that this rapid and facile LbL synthesis of multilayered hydrogels and technique for in situ loading with PTX, where the location of loading controls the release pattern, will find applications in biomedicine and pharmaceutics as a promising new technique. PMID- 19560819 TI - Anandamide-induced cell death: dual effects in primary rat decidual cell cultures. AB - Anandamide (AEA) belongs to an emerging class of lipid mediators collectively termed "endocannabinoids". This endogenously synthesized compound has been implicated in multiple processes, mainly related to the regulation of cell growth/death. During pregnancy endometrial fibroblast-like stromal cells proliferate and differentiate into decidual cells, forming the decidua. After reaching its maximum development, the decidua undergoes regression, which appears to be associated with apoptosis. In order to study the role of this endocannabinoid in this process, the effects of AEA upon cell viability and cell death in primary rat decidual cell cultures was investigated. The results obtained demonstrated that AEA induces cell death, in a dose-dependent manner which is associated with morphological alterations, such as nuclear condensation, DNA fragmentation and upregulation of caspase-3/7 activities. Moreover, these effects were attenuated by AM251, a selective antagonist for the cannabinoid receptor CB1. High concentrations induced a dramatic effect in cell viability and morphology, though methyl-beta-cyclodextrin (MCD), a membrane cholesterol depletor completely reversed the cytotoxic effect. These findings suggest that AEA in the uterine environment may play an important role in regulating apoptosis through CB1 and thereby modulate decidual stability and regression during pregnancy. However, it cannot be discarded the hypothesis that AEA, in high concentrations, represent a deleterious factor during this complex process. PMID- 19560820 TI - Effects of pulse duration and post-exposure period on the nitrite toxicity to a freshwater amphipod. AB - This research assesses the effects of nitrite pulses and post-exposure periods after nitrite exposures on the survival of the freshwater amphipod Eulimnogammarus toletanus. A toxicity bioassay was performed using three different nitrite concentrations (0.5, 5.0 and 10.0 mg/L NO(2)-N), four pulse exposures (1, 8, 24 and 48 h) for each nitrite concentration, and four post exposure times until to complete 96 h (i.e., 95, 88, 72 and 48 h, respectively). Our results showed a significant effect of nitrite concentrations, pulses and post-exposure times on the mortality of E. toletanus. The cumulative mortality at the end of pulse and that at the end of post-exposure time (delayed mortality) were different. We conclude that due to the high frequency of intermittent pollution in aquatic ecosystems it is necessary to incorporate the post-exposure effects into the traditional toxicological parameters to achieve a more realistic assessment of toxicants, especially at very short-term exposures. PMID- 19560821 TI - Revisiting the definition of a system of care: a sign of health or confusion? PMID- 19560822 TI - Textiloma: an uncommon complication of posterior lumbar surgery. AB - A surgical sponge or cotton swab that is inadvertently left behind in a surgical wound eventually becomes a "textiloma". Such foreign material (also called "gossypiboma") can cause a foreign-body reaction in the surrounding tissue. Textiloma is mostly asymptomatic in chronic cases, but can be confused with other soft-tissue masses. Therefore, it is important to be aware of patients who present with a paraspinal soft-tissue mass and unusual or atypical symptoms. Imaging is helpful for arriving at the correct diagnosis. Here, we describe a case of textiloma in which the patient presented with low-back pain 6 years after laminectomy and lumbar discectomy. Spinal computed tomography (CT) and magnetic resonance imaging (MRI) revealed a mass lesion in the posterior paravertebral region. PMID- 19560823 TI - [An unusual MRI observation of neurosarcoidosis]. AB - Sarcoidosis is a multisystemic granulomatous disease, characterized by the formation of an inflammatory lesion, the noncaseating giant-cell granuloma. Neurosarcoidosis represents only 5% of cases, but this figure is probably an underestimation. Diagnosis is difficult because of its clinical and radiological polymorphism. Magnetic resonance imaging (MRI) represents the "gold standard" of central nervous system exploration. We report here on an atypical example of primary neurosarcoidosis, according to MRI semiology, with preferential involvement of the Virchow-Robin perivascular spaces. PMID- 19560824 TI - Proteins accessible to immune surveillance show significant T-cell epitope depletion: Implications for vaccine design. AB - T cell activation is the final step in a complex pathway through which pathogen derived peptide fragments can elicit an immune response. For it to occur, peptides must form stable complexes with Major Histocompatibility Complex (MHC) molecules and be presented on the cell surface. Computational predictors of MHC binding are often used within in silico vaccine design pathways. We have previously shown that, paradoxically, most bacterial proteins known experimentally to elicit an immune response in disease models are depleted in peptides predicted to bind to human MHC alleles. The results presented here, derived using software proven through benchmarking to be the most accurate currently available, show that vaccine antigens contain fewer predicted MHC binding peptides than control bacterial proteins from almost all subcellular locations with the exception of cell wall and some cytoplasmic proteins. This effect is too large to be explained from the undoubted lack of precision of the software or from the amino acid composition of the antigens. Instead, we propose that pathogens have evolved under the influence of the host immune system so that surface proteins are depleted in potential MHC-binding peptides, and suggest that identification of a protein likely to contain a single immuno-dominant epitope is likely to be a productive strategy for vaccine design. PMID- 19560825 TI - Cost-effectiveness analysis of intracameral cefuroxime use for prophylaxis of endophthalmitis after cataract surgery. AB - OBJECTIVE: To evaluate the cost-effectiveness of intracameral cefuroxime for postoperative endophthalmitis prophylaxis, and to determine the efficacy threshold necessary for alternative antibiotics to attain cost-effective equivalence with intracameral cefuroxime. DESIGN: Cost-effectiveness analysis. PARTICIPANTS: We study a hypothetical cohort of 100,000 patients undergoing cataract surgery as a part of the cost analysis. METHODS: A cost-effectiveness model was constructed to analyze different antibiotic prophylactic regimens for postoperative endophthalmitis with intracameral cefuroxime as our base case. Efficacy was defined as the absolute reduction in rate of infection from background rate of infection, which was sourced from the literature. Antibiotic cost data were derived from the Red Book 2007 edition, and salary data were taken from the United States Bureau of Labor Statistics. Multivariate sensitivity analysis assessed the performance of antibiotic options under different scenarios. MAIN OUTCOME MEASURES: Cost per case of endophthalmitis prevented; theoretical maximal cost-effectiveness; efficacy threshold necessary to achieve cost-effective equivalence with intracameral cefuroxime; ratio indicating how many times more effective or less expensive alternative antibiotics would have to be to achieve cost-effective equivalence with intracameral cefuroxime. RESULTS: The cost-effectiveness ratio for intracameral cefuroxime is $1403 per case of postoperative endophthalmitis prevented. By comparison, the least expensive topical fluoroquinolone in our study, ciprofloxacin, would have to be >8 times more effective than intracameral cefuroxime to achieve cost-effective equivalence. The most expensive topical fluoroquinolones studied, gatifloxacin and moxifloxacin, would have to be > or =19 times more effective than intracameral cefuroxime to achieve cost-effective equivalence. A sensitivity analysis reveals that even in the worst case scenario for intracameral cefuroxime efficacy and with a 50% reduction in the cost of 4th-generation fluoroquinolones, gatifloxacin and moxifloxacin would have to be > or =9 times more effective than intracameral cefuroxime to achieve cost-effective equivalence. CONCLUSIONS: Administration of intracameral cefuroxime is relatively cost-effective in preventing endophthalmitis after cataract surgery. Owing to their high costs, many commonly used topical antibiotics are not cost-effective compared with intracameral cefuroxime, even under optimistic assumptions about their efficacy. PMID- 19560826 TI - Do infants influence their quality of care? Infants' communicative gestures predict caregivers' responsiveness. AB - Infants' effects on adults are a little studied but important aspect of development. What do infants do that increases caregiver responsiveness in childcare environments? Infants' communicative behaviors (i.e. smiling, crying) affect mothers' responsiveness; and preschool children's language abilities affect teachers' responses in the classroom setting. However, the effects of infants' intentional communications on either parents' or non-parental caregivers' responsiveness have not been examined. Using longitudinal video data from an infant classroom where infant signing was used along with conventional gestures (i.e. pointing), this study examines whether infants' use of gestures and signs elicited greater responsiveness from caregivers during daily interactions. Controlling child age and individual child effects, infants' gestures and signs used specifically to respond to caregivers elicited more responsiveness from caregivers during routine interactions. Understanding the effects of infants' behaviors on caregivers is critical for helping caregivers understand and improve their own behavior towards children in their care. PMID- 19560827 TI - Onset-age of bipolar disorders at six international sites. AB - BACKGROUND: Onset-age is a stable characteristic of bipolar disorder (BPD) patients of clinical and probable psychobiological importance, but large pooled clinical samples from multiple sites employing modern diagnostic criteria to quantify onset-age remain rare. METHODS: We pooled diagnostic, demographic, and clinical data from 1566 BPD patients from six international sites (5 European, 1 US) to compare onset-ages in subgroups. RESULTS: Median+/-IQR onset in 1090 BP-I patients was 5.8 years younger than 476 BP-II cases (24.3+/-18.3 vs. 30.1+/-13.8 years; p<0.0001). Onset-age ranked: [a] BP-I men (23.0+/-12.8); [b] BP-I women (26.0+/-14.2); [c] BP-II men (29.7+/-19.1); and [d] BP-II women (30.1+/-17.5 years. Juvenile-onset (or=10% within 3 months is the use of very-low-calorie diets (VLCDs). In patients with diabetes, VLCDs are associated with rapid improvement in glycaemia and cardiovascular risk factors. The present review analyses the evidence from available trials on the effects of VLCDs on body weight, glycaemic control and complications, and their potential for clinical use in diabetes management. PMID- 19560837 TI - Evolution is intelligent design. PMID- 19560836 TI - Expression of Bcl-2 predicts outcome in locally advanced non-small cell lung cancer patients treated with cisplatin-based concurrent chemoradiotherapy. AB - BACKGROUND: Platinum-based concurrent chemoradiotherapy (CCRT) is a standard treatment for locally advanced unresectable non-small cell lung cancer (NSCLC). The determination of parameters that may predict the result of the treatment has strong clinical implications. PATIENTS AND METHODS: Pretreatment tumor biopsy specimens from 39 patients with locally advanced NSCLC (stage IIIA: 5, stage IIIB: 34) were analyzed for p53, Bcl-2, Bax and ERCC1 expression by immunohistochemistry. All patients were treated with cisplatin-based CCRT. Twenty four patients received induction chemotherapy followed by CCRT (60Gy/30 fractions, 6mg/m(2) of cisplatin daily). The most commonly administered induction chemotherapy regimen was VIP (etoposide, ifosfamide, cisplatin; 20 patients). Fifteen patients received the same CCRT without induction chemotherapy. RESULTS: High expression of p53, Bcl-2, Bax and ERCC1 was observed in 15 (38%), 19 (49%), 17 (44%) and 12 (31%) patients, respectively. High expression of Bcl-2 was significantly associated with longer survival duration (20 months vs. 9 months, P=0.008) and better response to the treatment (74% vs. 30%, P=0.01). In multivariate analysis, Bcl-2 expression was the only significant independent prognostic factor of overall survival (P=0.007) among the pretreatment patients characteristics. CONCLUSIONS: High expression of Bcl-2 may be a useful prognostic factor in locally advanced NSCLC patients treated with cisplatin-based CCRT. PMID- 19560838 TI - Randomized clinical trial comparing a patient-driven titration protocol of intravenous hydromorphone with traditional physician-driven management of emergency department patients with acute severe pain. AB - STUDY OBJECTIVE: We test the null hypothesis that the "1+1" hydromorphone patient driven protocol is clinically and statistically equivalent in safety and efficacy to that of traditional physician-driven administration of opioids for emergency department (ED) treatment of acute severe pain. METHODS: This was a prospective randomized clinical trial of nonelderly adults presenting to an urban academic ED with acute pain of sufficient severity to warrant intravenous (IV) opioids in the judgment of the attending physician. Patients randomized to the 1+1 hydromorphone patient-driven protocol received 1 mg IV hydromorphone followed by a second 1-mg dose 15 minutes later if the patient responded affirmatively to the question, "Do you want more pain medication?" Patients in the physician-driven group received any IV opioid in the dose chosen by the ED attending physician, with any additional analgesia provided at the discretion of that physician. The primary outcome was the difference in improvement in pain between the 2 groups at 60 minutes, as measured by a validated and reproducible numeric rating scale. Secondary outcomes included incidence of oxygen desaturation, hypoventilation, hypotension, bradycardia, nausea, vomiting, pruritus, and use of naloxone. RESULTS: The mean decrease in numeric rating scale pain scores for the 1+1 hydromorphone patient-driven group was 5.6 versus 4.5 in the physician-driven group. The difference of 1.1 numeric rating scale units (95% confidence interval 0.3 to 1.9) was statistically significant but fell 0.2 numeric rating scale units short of the 1.3 numeric rating scale unit threshold required to attain clinically significant efficacy. Safety profiles were similarly satisfactory in both groups. Ninety-four percent of the 1+1 hydromorphone patient-driven group achieved adequate analgesia (as defined by the patient) within 60 minutes of protocol initiation. CONCLUSION: The 1+1 hydromorphone patient-driven protocol is statistically superior and at least as clinically efficacious and safe as traditional physician-driven treatment of ED patients with acute severe pain. More than 9 of 10 patients randomized to the study protocol achieved satisfactory pain control, as defined by the patient, within an hour or less. PMID- 19560840 TI - [Community medicine and mental health]. PMID- 19560841 TI - [Repeat sick leave procedures and diagnosis]. AB - OBJECTIVE: To find out the recurrence of absence due to sick leave from the perspective of diagnosing the disease-causing absenteeism. DESIGN: Case-control study. CONTROLS: workers who only had only been on sick leave once. CASES: workers with repeated sick leave. CASES with 2, 3, 4 or more absences due to sick leave were analysed separately. SETTING: Guipuzcoa, from 2001 to 2005. PARTICIPANTS: There were 7,313 sick leave procedures collected from a Mutual Company with the medical reason, age and gender were duly registered and coded. Of the 6,030 workers who took some time off work, 5,016 had a single sick leave procedure, and were considered as controls and there were 1,014 cases that had two or more sick leave procedures. MEASUREMENTS: We calculated the odds ratio (OR) and 95% confidence intervals, adjusting for age and stratified by gender, using a logistic regression model. The slope of sick leave repeats was evaluated. RESULTS: An increase in the risk of recurrent sick leave was observed for both genders over the age of 45, with a significant slope. There was an increase in the risk of recurrent sick leave in the most common diseases, musculoskeletal, external causes, mental and gastrointestinal in males, and pregnancy, musculoskeletal, mental and external causes in females. CONCLUSIONS: An alternative method for verifying the correct prescribing of sick leave would be to just study diseases that occur most often. This would focus on those workers who had recurrent absences due to sick leave. PMID- 19560839 TI - Regional age-related effects in the monkey brain measured with 1H magnetic resonance spectroscopy. AB - The rhesus monkey is a useful model for examining age-related effects on the brain, because of the extensive neuroanatomical homology between the monkey and the human brain, the tight control for neurological diseases as well as the possibility of obtaining relevant behavioral data and post-mortem tissue for histological analyses. Here, proton magnetic resonance spectroscopy ((1)H-MRS) was used together with high-resolution anatomical MRI images to carefully assess regional concentrations of brain metabolites in a group of 20 rhesus monkeys. In an anterior volume of interest (VOI) that covered frontal and prefrontal areas, significant positive correlations of myo-inositol and of total creatine concentrations with age were detected, whereas N-acetyl aspartate (NAA) and choline compounds (Cho) were not significantly correlated with age. In an occipito-parietal VOI, all metabolites showed no statistically significant age dependent trend. Strong correlations were found between NAA concentration and gray matter fraction in the VOIs as well as between choline compounds and white matter fraction. PMID- 19560842 TI - Synthesis of substituted-phenyl-1,2,4-triazol-3-thione analogues with modified D glucopyranosyl residues and their antiproliferative activities. AB - A series of D-glucopyranosyl-1,2,4-triazole-3-thione derivatives 1a-1d were synthesized by the reaction of 1,2,4-triazole-3-thione Schiff bases 5a-5d with 2,3,4,6-tetra-O-acetyl-sigma-D-glucopyranosyl bromide. We demonstrate the conversion of 2 to 5, without the necessity of purification of both oxadiazole and triazole intermediates to afford the compounds 5. Their structures were confirmed by standard studies of (1)H NMR, IR, MS and elemental analysis. Analogues 5 and 1 have shown cytotoxic activity against human MCF-7 and Bel-7402 malignant cell lines. PMID- 19560843 TI - Design, synthesis and in vitro antibacterial/antifungal evaluation of novel 1 ethyl-6-fluoro-1,4-dihydro-4-oxo-7(1-piperazinyl)quinoline-3-carboxylic acid derivatives. AB - A series of 1-ethyl-6-fluoro-1,4-dihydro-4-oxo-7(1-piperazinyl)quinoline-3 carboxylic acid (norfloxacin) derivatives were prepared according to the principle of combinating bioactive substructures and tested for their activities against five plant pathogenic bacteria and three fungi in vitro. The preliminary bioassays indicated that almost all synthesized target compounds retained the antibacterial activities of norfloxacin and had some antifungal activities as carboxylic acid amide compounds. The activities of compounds 1 and 22 against Xanthomonas oryzae were better than norfloxacin and all tested compounds had better antibacterial activities as compared to the agricultural streptomycin sulfate (a commercial bactericide) against X. oryzae, Xanthomonas axonopodis and Erwinia aroideae. Additionally, compounds 2 and 20 displayed good antifungal activities against Rhizoctonia solani and their inhibition of growth reached 83% and 94% respectively at the concentration of 200mg/L. PMID- 19560844 TI - How to be a professor: what the books don't tell you. PMID- 19560845 TI - Radiological risk assessment and biosphere modelling for radioactive waste disposal in Switzerland. AB - Long-term safety assessments for geological disposal of radioactive waste in Switzerland involve the demonstration that the annual radiation dose to humans due to the potential release of radionuclides from the waste repository into the biosphere will not exceed the regulatory limit of 0.1 mSv. Here, we describe the simple but robust approach used by Nagra (Swiss National Cooperative for the Disposal of Radioactive Waste) to quantify the dose to humans as a result to time dependent release of radionuclides from the geosphere into the biosphere. The model calculates the concentrations of radionuclides in different terrestrial and aquatic compartments of the surface environment. The fluxes of water and solids within the environment are the drivers for the exchange of radionuclides between these compartments. The calculated radionuclide concentrations in the biosphere are then used to estimate the radiation doses to humans due to various exposure paths (e.g. ingestion of radionuclides via drinking water and food, inhalation of radionuclides, external irradiation from radionuclides in soils). In this paper we also discuss recent new achievements and planned future work. PMID- 19560846 TI - Pteris vittata - revisited: uptake of As and its speciation, impact of P, role of phytochelatins and S. AB - Pteris vittata is known to hyperaccumulate As but the mechanism is poorly understood. We found an increase of As concentration with increasing soil solution As concentrations, but P application had no impact, although plant P concentrations responded to different rates of P supply. As in fronds was dominantly (82-89%) present in the form of AsIII. In roots we detected 45% as AsIII which is higher than reported in previous studies and supports substantial As-reduction to take place in roots. We detected PC2/3GS-AsIII, PC2-GS-AsIII and (PC2)2-AsIII in increasing amounts with application of As. The total amount of PC was in the range reported previously and far too small to assign a significant role in As detoxification to PCs. The close correlation between S and As in fronds and the lack of data on sulphur uptake and metabolism indicates the need for a detailed investigation on sulphur nutritional status and As metabolism in P. vittata. PMID- 19560847 TI - Environmental pollution by antibiotics and by antibiotic resistance determinants. AB - Antibiotics are among the most successful drugs used for human therapy. However, since they can challenge microbial populations, they must be considered as important pollutants as well. Besides being used for human therapy, antibiotics are extensively used for animal farming and for agricultural purposes. Residues from human environments and from farms may contain antibiotics and antibiotic resistance genes that can contaminate natural environments. The clearest consequence of antibiotic release in natural environments is the selection of resistant bacteria. The same resistance genes found at clinical settings are currently disseminated among pristine ecosystems without any record of antibiotic contamination. Nevertheless, the effect of antibiotics on the biosphere is wider than this and can impact the structure and activity of environmental microbiota. Along the article, we review the impact that pollution by antibiotics or by antibiotic resistance genes may have for both human health and for the evolution of environmental microbial populations. PMID- 19560848 TI - Renal resistance index in renal allograft recipients: a role for ADMA. AB - BACKGROUND: Renal resistance index, a predictor of kidney allograft function and patient survival, seems to depend on renal and peripheral vascular compliance and resistance. Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase and therefore influences vascular resistance. STUDY DESIGN: We investigated the relationship between renal resistance index, ADMA, and risk factors for cardiovascular diseases and kidney function in a cross-sectional study. SETTING & PARTICIPANTS: 200 stable renal allograft recipients (133 men and 67 women with a mean age of 52.8 years). PREDICTORS: Serum ADMA concentration, pulse pressure, estimated glomerular filtration rate and recipient age. OUTCOME: Renal resistance index. MEASUREMENTS: Renal resistance index measured by color coded duplex ultrasound, serum ADMA concentration measured by liquid chromatography-tandem mass spectrometry, estimated glomerular filtration rate (Nankivell equation), arterial stiffness measured by digital volume pulse, Framingham and other cardiovascular risk factors, and evaluation of concomitant antihypertensive and immunosuppressive medication. RESULTS: Mean serum ADMA concentration was 0.72 +/- 0.21 (+/-SD) micromol/L and mean renal resistance index was 0.71 +/- 0.07. Multiple stepwise regression analysis showed that recipient age (P < 0.001), pulse pressure (P < 0.001), diabetes (P < 0.01) and ADMA concentration (P < 0.01) were independently associated with resistance index. ADMA concentrations were correlated with estimated glomerular filtration rate (P < 0.01). LIMITATIONS: The cross-sectional nature of this study precludes cause-effect conclusions. CONCLUSIONS: In addition to established cardiovascular risk factors, ADMA appears to be a relevant determinant of renal resistance index and allograft function and deserves consideration in prospective outcome trials in renal transplantation. PMID- 19560850 TI - Anion and osmolal gaps after alcohol intoxication. PMID- 19560851 TI - The value of machine perfusion in deceased donor kidney transplantation. PMID- 19560849 TI - Cinacalcet-induced leukocytoclastic vasculitis. AB - An 80-year-old woman on maintenance hemodialysis therapy developed severe hypercalcemia under vitamin D treatment for secondary hyperparathyroidism. To avoid the toxic calcemic effects, cinacalcet was introduced and the dose of vitamin D was substantially decreased. Cinacalcet targets the calcium-sensing receptor and decreases parathyroid hormone levels without increasing calcium and phosphorus levels. Three days after starting cinacalcet therapy, the patient developed palpable purpura on both upper and lower extremities that resolved after discontinuation of cinacalcet and administration of steroids. Skin biopsy of the initial eruption showed leukocytoclastic vasculitis. According to the Naranjo adverse drug reaction probability scale, leukocytoclastic vasculitis probably was caused by cinacalcet introduction. Drug-induced vasculitis is a poorly defined disorder, and, in most cases, no pathogenetic mechanism can be described. An idiosyncratic reaction to the agent often is proposed. Cinacalcet should be considered a causative agent of cutaneous leukocytoclastic vasculitis, and although this is the result of only a clinical observation, further attention is required in the future because cinacalcet recently has been introduced in the treatment of secondary hyperparathyroidism in patients on long-term hemodialysis therapy. PMID- 19560852 TI - Association of level of kidney function and platelet aggregation in acute myocardial infarction. AB - BACKGROUND: Decreased kidney function has been established as an important risk factor in patients presenting with acute coronary syndrome. In acute coronary syndrome, increased platelet aggregation is associated with vascular complications. The aim of this study is to examine whether decreased kidney function is associated with altered platelet function in patients presenting with acute myocardial infarction. STUDY DESIGN: Prospective cohort. SETTING & PARTICIPANTS: 413 patients presenting with acute myocardial infarction admitted to the cardiac intensive care unit at Ostersund Hospital, Ostersund, Sweden. PREDICTORS: Glomerular filtration rate less than 60 mL/min/1.73 m(2) estimated from serum cystatin C level, comorbidity, medications, and markers of inflammation and hemostasis. OUTCOMES & MEASUREMENTS: Platelet aggregation was assessed by measuring the formation of small platelet aggregates (SPAs) by using a laser light scattering method. A greater SPA level indicates greater platelet aggregation. Platelet aggregation analysis was performed on days 1, 2, 3, and 5 in-hospital. RESULTS: We observed a significant increase in platelet aggregation during the first 3 days in the hospital regardless of kidney function (P < 0.001). Platelet aggregation was more pronounced in patients with estimated glomerular filtration rate less than 60 mL/min/1.73 m(2) on day 2 (SPA count, 65,000 versus 47,000; P = 0.01) and day 3 (SPA count, 77,000 versus 52,000; P = 0.02). In a multiple linear regression analysis, decreased kidney function was no longer significantly associated with increased platelet aggregation. Older age, greater plasma fibrinogen level, and diabetes mellitus were associated with increased platelet aggregation in the multivariable model. LIMITATIONS: During the study period, 78 patients presenting with acute myocardial infarction were not eligible for inclusion. Differences in treatment with antiplatelet medication between the 2 groups might have affected our findings. CONCLUSIONS: Platelet aggregation increases during the first days after acute myocardial infarction regardless of kidney function. There is no difference in platelet aggregation in patients according to level of kidney function. PMID- 19560853 TI - In utero tobacco exposure: the effects of heavy and very heavy smoking on the rate of SGA infants in the Federal State of Saarland, Germany. AB - OBJECTIVE: To assess the effects of heavy and very heavy smoking on the rate of small for gestational age (SGA) infants, and to assess socio-economic and regional differences in smoking patterns in pregnant women in Germany. STUDY DESIGN: The Neonatal and Perinatal database of the federal state of Saarland, Germany was used to perform a population-based analysis of preterm (>32 weeks of gestation) and term (>36 weeks of gestation) newborns in 2004-2006. The rate of SGA babies dependent on the amount of tobacco exposure among self-identified smokers and non-smokers were assessed, and distinct maternal risk factors for smoking were evaluated. Our data were compared with the German National Perinatal database. RESULTS: 14,593 paired data sets (peripartum/perinatal) were included in this study. The overall rate of smoking during pregnancy was 11.8% with a high percentage of pregnant women smoking 11-20 cigarettes/day (heavy smoker; 4.0%), and >20 cigarettes/day (very heavy smoker; 0.6%). Self-identified heavy tobacco use significantly increased the risk for SGA infants (p<0.01) in women without uteroplacental insufficiency. Risk factors for smoking included ethnicity (German/Caucasian), socio-economic parameters (single vs. non-single households, status of employment) and age. Smoking pattern and the rate of SGA babies in our cohort differed substantially from the national average. CONCLUSIONS: Although the overall rate of smoking appears comparable to previously published data, heavy and very heavy smoking was high in our cohort. Heavy smoking was disproportionately associated with SGA. Preventative measures and strategies should take into consideration socio-economic risk factors as well as regional differences, and should be targeted at distinct subgroups that are especially prone to smoking during pregnancy. PMID- 19560854 TI - Poly-l-lysines and poly-l-arginines induce leakage of negatively charged phospholipid vesicles and translocate through the lipid bilayer upon electrostatic binding to the membrane. AB - Poly-l-lysines (PLL) and poly-l-arginines (PLA) of different polymer chain lengths interact strongly with negatively charged phospholipid vesicles mainly due to their different electrical charges. 1-Palmitoyl-2-oleoyl-sn-glycero-3 phosphoglycerol (POPG), 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) and their mixtures (1/1 mol/mol) with the respective phosphatidylcholines of equivalent chain length were chosen as model membrane systems that form at room temperature either the fluid L(alpha) or the gel phase L(beta) lipid bilayer membranes, respectively. Leakage experiments revealed that the fluid POPG membranes are more perturbed compared to the gel phase DPPG membranes upon peptide binding. Furthermore, it was found that pure PG membranes are more prone to release the vesicle contents as a result of pore formation than the lipid mixtures POPG/POPC and DPPG/DPPC. For the longer polymers (>or=44 amino acids) maximal dye-release was observed when the molar ratio of the concentrations of amino acid residues to charged lipid molecules reached a value of R(P)=0.5, i.e. when the outer membrane layer was theoretically entirely covered by the polymer. At ratios lower or higher than 0.5 leakage dropped significantly. Furthermore, PLL and PLA insertions and/or translocations through lipid membranes were analyzed by using FITC-labeled polymers by monitoring their fluorescence intensity upon membrane binding. Short PLL molecules and PLA molecules of all lengths seemed to translocate through both fluid and gel phase lipid bilayers. Comparison of the PLL and PLA fluorescence assay results showed that PLA interacts stronger with phospholipid membranes compared to PLL. Isothermal titration calorimetry (ITC) measurements were performed to give further insight into these mechanisms and to support the findings obtained by fluorescence assays. Cryo-transmission electron microscopy (cryo-TEM) was used to visualize changes in the vesicles' morphology after addition of the polypeptides. PMID- 19560856 TI - An ex vivo study of the correlation between acoustic emission and microvascular damage. AB - The objective of this study was to conduct an ex vivo examination of correlation between acoustic emission and tissue damage. Intravital microscopy was employed in conjunction with ultrasound exposure in cremaster muscle of male Wistar rats. Definity microbubbles were administered intravenously through the tail vein (80microL.kg(-1).min(-1)infusion rate) with the aid of a syringe pump. For the pulse repetition frequency (PRF) study, exposures were performed at four locations of the cremaster at a PRF of 1000, 500, 100 and 10Hz (one location per PRF per rat). The 100-pulse exposures were implemented at a peak rarefactional pressure (P(r)) of 2MPa, frequency of 2.25MHz with 46 cycle pulses. For the pressure amplitude threshold study, 100-pulse exposures (46 cycle pulses) were conducted at various peak rarefactional pressures from 0.5MPa to 2MPa at a frequency of 2.25MHz and PRF of 100Hz. Photomicrographs were captured before and 2-min postexposure. On a pulse-to-pulse basis, the 10Hz acoustic emission was considerably higher and more sustained than those at other PRFs (1000, 500, and 100Hz) (p<0.05). Damage, measured as area of extravasation of red blood cells (RBCs), was also significantly higher at 10Hz PRF than at 1000, 500 and 100Hz (p<0.01). The correlation of acoustic emission to tissue damage showed a trend of increasing damage with increasing cumulative function of the relative integrated power spectrum (CRIPS; R(2)=0.75). No visible damage was present at P(r)< or =0.85MPa. Damage, however, was observed at P(r)> or =1.0MPa and it increased with increasing acoustic pressure. PMID- 19560857 TI - Castration-resistant prostate cancer: from new pathophysiology to new treatment targets. AB - CONTEXT: Castration-resistant prostate cancer (CRPC) refers to patients who no longer respond to surgical or medical castration. Standard treatment options are limited. OBJECTIVE: To review the concepts and rationale behind targeted agents currently in late-stage clinical testing for patients with CRPC. EVIDENCE ACQUISITION: Novel targeted therapies in clinical trials were identified from registries. The MEDLINE database was searched for all relevant reports published from 1996 to October 2009. Bibliographies of the retrieved articles and major international meeting abstracts were hand-searched to identify additional studies. EVIDENCE SYNTHESIS: Advances in our understanding of the molecular mechanisms underlying prostate cancer (PCa) progression has translated into a variety of treatment approaches. Agents targeting androgen receptor (AR) activation and local steroidogenesis, angiogenesis, immunotherapy, apoptosis, chaperone proteins, the insulin-like growth factor (IGF) pathway, RANK-ligand, endothelin receptors, and the Src family kinases are entering or have recently completed accrual to phase 3 trials for patients with CRPC. CONCLUSIONS: A number of new agents targeting mechanisms of PCa progression with early promising results are in clinical trials and have the potential to provide novel treatment options for CRPC in the near future. PMID- 19560855 TI - Activation of RXR and RAR signaling promotes myogenic differentiation of myoblastic C2C12 cells. AB - Differentiation of embryonic and adult myogenic progenitors undergoes a complex series of cell rearrangements and specification events which are controlled by distinct gene regulatory networks. Delineation of the molecular mechanisms that regulate skeletal muscle specification and formation should be important for understanding congenital myopathies and muscular degenerative diseases. Retinoic acid (RA) signaling plays an important role in development. However, the role of RA signaling in adult myogenic progenitors is poorly understood. Here, we investigate the role of RA signaling in regulating myogenic differentiation of myoblastic progenitor cells. Using the mouse myoblast progenitor C2C12 line as a model, we have found that the endogenous expression of most RAR and RXR isotypes is readily detected. While the nuclear receptor co-repressors are highly expressed, two of the three nuclear receptor co-activators and the enzymes involved in RA synthesis are expressed at low level or undetectable, suggesting that the RA signaling pathway may be repressed in myogenic progenitors. Using the alpha-myosin heavy chain promoter-driven reporter (MyHC-GLuc), we have demonstrated that either ATRA or 9CRA is able to effectively induce myogenic differentiation, which can be synergistically enhanced when both ATRA and 9CRA are used. Upon ATRA and 9CRA treatment of C2C12 cells the expression of late myogenic markers significantly increases. We have further shown that adenovirus mediated exogenous expression of RARalpha and/or RXRalpha is able to effectively induce myogenic differentiation in a ligand-independent fashion. Morphologically, ATRA- and 9CRA-treated C2C12 cells exhibit elongated cell body and become multi nucleated myoblasts, and even form myoblast fusion. Ultrastructural analysis under transmission electron microscope reveals that RA-treated myogenic progenitor cells exhibit an abundant presence of muscle fibers. Therefore, our results strongly suggest that RA signaling may play an important role in regulating myogenic differentiation. PMID- 19560858 TI - Refractory hematuria in an oliguric patient after pancreas transplantation with exocrine pancreas bladder drainage. AB - A 52-yr-old man presented with hematuria and clot retention. He had undergone simultaneous pancreas-kidney transplantation with exocrine pancreas bladder drainage 16 yr ago. The patient suffered from progressive transplant kidney failure with gradually decreasing urine output and needed hemodialysis every other day. Gross hematuria persisted after removal of all blood clots. Cystoscopy showed multiple small, flat ulcers of the bladder mucosa. Some bled discretely and were coagulated cautiously. However, hematuria was refractory to multiple urological interventions, which eventually necessitated an enteric diversion of the exocrine pancreas. Hematuria ceased following an uneventful postoperative course. PMID- 19560860 TI - Medical therapy to facilitate the passage of stones: what is the evidence? AB - CONTEXT: Medical expulsive therapy (MET) for urolithiasis has gained increasing attention in the last years. It has been suggested that the administration of alpha-adrenoreceptor antagonists (alpha-blockers) or calcium channel blockers augments stone expulsion rates and reduces colic events. OBJECTIVE: To evaluate the efficacy and safety of MET with alpha-blockers and calcium channel blockers for upper urinary tract stones with and without prior extracorporeal shock wave lithotripsy (ESWL). EVIDENCE ACQUISITION: A systematic review of the literature was performed in Medline, Embase, the Cochrane Central Register of Controlled Trials, and the Cochrane Database of Systematic Reviews searched through 31 December 2008 without time limit. Efficacy and safety end points were evaluated in 47 randomised, controlled trials assessing the role of MET. Meta-analysis was conducted using Review Manager (RevMan) v.5.0 (The Nordic Cochrane Centre, The Cochrane Collaboration, Copenhagen, Denmark). EVIDENCE SYNTHESIS: Pooling of alpha-blocker and calcium channel blocker studies demonstrated a higher and faster expulsion rate compared to a control group (risk ratio [RR]: 1.45 vs 1.49; 95% confidence interval [CI]: 1.34-1.57 vs 1.33-1.66). Similar results have been obtained after ESWL (RR: 1.29 vs 1.57; 95% CI: 1.16-1.43 vs 1.21-2.04). Additionally, lower analgesic requirements, fewer colic episodes, and fewer hospitalisations were observed within treatment groups. CONCLUSIONS: Pooled analyses suggest that MET with alpha-blockers or calcium channel blockers augments stone expulsion rates, reduces the time to stone expulsion, and lowers analgesia requirements for ureteral stones with and without ESWL for stones < or = 10 mm. There is some evidence that a combination of alpha-blockers and corticosteroids might be more effective than treatment with alpha-blockers alone. Renal stones after ESWL also seem to profit from MET. The vast majority of randomised studies incorporated into the present systematic review are small, single-centre studies, limiting the strength of our conclusions. Therefore, multicentre, randomised, placebo-controlled trials are needed. PMID- 19560859 TI - Open to debate. The motion: Robotic partial nephrectomy is better than open partial nephrectomy. PMID- 19560861 TI - Hurdles in modern trial management: as overbureaucracy and commercialization of clinical trials suffocate independent academic research, it's time for a change. PMID- 19560862 TI - Butein suppresses c-Myc-dependent transcription and Akt-dependent phosphorylation of hTERT in human leukemia cells. AB - Telomerase, a ribonucleoprotein that plays an important role in neoplastic immortality, is up-regulated in approximately 85% of cancers, especially in leukemia. The polyphenol, butein, has potent effects against various types of cancer cells, but its effects on telomerase activity have not been well characterized. In this study, we show that butein causes a down-regulation of hTERT gene expression and a concomitant decrease of telomerase activity. Butein also suppresses expression of c-Myc at the transcriptional level and down regulates DNA-binding activity, regardless of cell type specificity, in leukemia cells. DNA-binding activities of c-Myc to the hTERT core promoter were decreased in butein-treated cells, as seen by chromatin immunoprecipitation assay. Treatment with butein also suppressed the activation of Akt, thereby inhibiting hTERT phosphorylation and translocation into the nucleus. In this process, butein also up-regulated the surface expression of CD11b in leukemia cells. Inhibition of telomerase activity by butein was followed by loss of proliferative capacity, induction of apoptosis, and differentiation. These findings demonstrate the effectiveness of butein at inhibiting telomerase activity by down-regulating hTERT gene expression in human leukemia cells. PMID- 19560863 TI - Collection of SiO2, Al2O3 and Fe2O3 particles using a gas-solid fluidized bed filter. AB - The filtration of SiO(2), Al(2)O(3) and Fe(2)O(3) particles with average sizes of 4 and 40 microm using a fluidized bed filter at 40 and 300 degrees C was studied. The collection mechanisms, interparticle forces and bounce-off effect between filtered particles and collectors were analyzed to determine their effect on particle filtration. Experimental results showed that the collection efficiency of 4 microm SiO(2) and Al(2)O(3) particles exceeded that of 40 microm particles. Contrarily, the 40 microm Fe(2)O(3) particles were collected more efficiently than the 4 microm particles, because of the differences between the microstructures of SiO(2), Al(2)O(3,) and Fe(2)O(3) particles. The interaction between the particles affected the removal of mixed SiO(2), Al(2)O(3) and Fe(2)O(3). The particle size distribution (PSD) of the particles in the exit was governed by the operating temperature, the original size of the filtered particles, the interparticle force and the hardness of the particles and the collectors. The smallest particles were not those most easily elutriated from the fluidized bed filter because they agglomerated with each other or with large particles. The van der Waal's force dominated the forces between 4 and 40 microm particles. The main collection mechanism for 4 and 40 microm particles was direct interception. The effect of impaction increased with particle size above 40 microm. The strong impaction and bounce-off effect reduced the collection efficiency of 40 microm SiO(2) and Al(2)O(3) particles. However, the strong interparticle force between Fe(2)O(3) particles and collectors contributed to the high collection efficiency of the Fe(2)O(3) particles. PMID- 19560864 TI - A life cycle assessment of destruction of ammunition. AB - The Swedish Armed Forces have large stocks of ammunition that were produced at a time when decommissioning was not considered. This ammunition will eventually become obsolete and must be destroyed, preferably with minimal impact on the environment and in a safe way for personnel. The aim of this paper is to make a comparison of the environmental impacts in a life cycle perspective of three different methods of decommissioning/destruction of ammunition, and to identify the environmental advantages and disadvantages of each of these destruction methods: open detonation; static kiln incineration with air pollution control combined with metal recycling, and a combination of incineration with air pollution control, open burning, recovery of some energetic material and metal recycling. Data used are for the specific processes and from established LCA databases. Recycling the materials in the ammunition and minimising the spread of airborne pollutants during incineration were found to be the most important factors affecting the life cycle environmental performance of the compared destruction methods. Open detonation with or without metal recycling proved to be the overall worst alternative from a life cycle perspective. The results for the static kiln and combination treatment indicate that the kind of ammunition and location of the destruction plant might determine the choice of method, since the environmental impacts from these methods are of little difference in the case of this specific grenade. Different methods for destruction of ammunition have previously been discussed from a risk and safety perspective. This is however to our knowledge the first study looking specifically on environmentally aspect in a life cycle perspective. PMID- 19560865 TI - Alpha-2-delta ligands in fibromyalgia: is the glass half empty or full? PMID- 19560866 TI - Activation of the prostaglandin system in response to sleep loss in healthy humans: potential mediator of increased spontaneous pain. AB - Insufficient duration of sleep is a highly prevalent behavioral pattern in society that has been shown to cause an increase in spontaneous pain and sensitivity to noxious stimuli. Prostaglandins (PGs), in particular PGE2, are key mediators of inflammation and pain, and we investigated whether PGE2 is a potential mediator in sleep-loss-induced changes in nociceptive processing. Twenty-four participants (7 females, age 35.1+/-7.1 years) stayed for 7 days in the Clinical Research Center. After two baseline days, participants were randomly assigned to either 3 days of 88 h of sleep deprivation (TSD, N=15) or 8h of sleep per night (N=9), followed by a night of recovery sleep. Participants rated the intensity of various pain-related symptoms every 2h across waking periods on computerized visual analog scales. PGE2 was measured in 24-h-urine collections during baseline and third sleep deprivation day. Spontaneous pain, including headache, muscle pain, stomach pain, generalized body pain, and physical discomfort significantly increased by 5-14 units on a 100-unit scale during TSD, compared to the sleep condition. Urinary PGE2 metabolite significantly increased by about 30% in TSD over sleep condition. TSD-induced increase in spontaneous pain, in particular headache and muscle pain, was significantly correlated with increase in PGE2 metabolite. Activation of the PGE2 system appears to be a potential mediator of increased spontaneous pain in response to insufficient sleep. PMID- 19560867 TI - Fabrication of various tip-size AFM probes for evaluating single-molecular retraction force between actin and anti-actin. AB - The authors fabricated a probe tip with various sizes and examined the size dependency of the probe tip on the distribution of retraction forces between actin and anti-actin. Probe tips of various sizes were fabricated by two-photon polymerization methods on a micro cantilever of an atomic force microscope (AFM). The authors succeeded in fabricating a spherical tip having a smooth surface and the tip size varied between phi 0.8 and 5.5 microm. Anti-actin was immobilized on the fabricated probe tips and force curves were measured against an actin immobilized mica substrate by AFM to analyze the retraction forces. The histograms of retraction forces showed that the single-molecular retraction force between actin and anti-actin was ca. 350-400 pN. It was observed that the average retraction forces for each tip size correlated with the square of the tip radius. PMID- 19560868 TI - Canada and veterinary parasitology. AB - A World Association for the Advancement of Veterinary Parasitology tradition for its conference is to present some highlights of the country hosting the event, and with an emphasis on the history of, and research in, veterinary parasitology. A review of Canada's peoples, physiography, climate, natural resources, agriculture, animal populations, pioneers in veterinary parasitology, research accomplishments by other veterinary parasitologists, centres for research in veterinary parasitology, and major current research had been presented at a World Association for the Advancement of Veterinary Parasitology Conference in Canada in 1987, and was published. The present paper updates the information on the above topics for the 22 years since this conference was last held in Canada. PMID- 19560869 TI - Parasite control in the age of drug resistance and changing agricultural practices. AB - The benefits of using antiparasitic drugs in farm animals are unquestionable. However, despite anthelmintic use as the predominant control strategy, extreme parasite infection cases are appearing in sheep and goat production; these impact productivity and have show mortality rates reaching pre-drug use levels. This was a predictable situation resulting from the loss of efficacy by all available products, particularly when some products were used as the sole intervention. The concepts of agroecology and holistic agriculture, which advocate the use of integrated management strategies, such as target selected treatment, herbal medicine, and the application of other parasite control alternatives, are not completely new, but are undergoing a resurgence because of their more sustainable appeal. The objective of this review article is to examine the problem of parasite control in the face of parasite drug resistance and to outline some strategies that may be used in parasite control programmes. Before they are accepted and recommended by the WAAVP, agroecological methods such as those listed above and described in detail herein should be validated based on scientific evidence of their efficacy for parasite control and should be tested for both host and environmental safety. PMID- 19560870 TI - Comments on the article of Ming-Hsien Li and Hong-Kean Ooi "Fecal occult blood manifestation of intestinal Eimeria spp. infection in rabbit" [Vet. Parasitol. 161 (2009) 327-329]. PMID- 19560871 TI - Assessing the safety and compatibility of silver based wound dressings in a magnetic resonance environment. AB - INTRODUCTION: Silver dressings are an integral part of the management of burn patients. Package inserts assert a lack of compatibility and safety with magnetic resonance imaging (MRI) and recommend removal prior to any MRI procedure, although there is no clear evidence to support this recommendation. Dressing removal is associated with increased pain, anxiety, stress, and analgesia use. This study was to determine whether these products produce MRI image distortion or if the agitation of the silver particles generates enough heat which might produce further skin damage. METHODS: Hind limbs from euthanized pigs were used in a 7T MRI scanner with three standard silver wound dressings. Images were obtained with both dry and wet dressings. Temperature was assessed before and during MRI by probes inserted between the dressing and skin. Images were independently reviewed by a radiologist and MR physicist for distortion. RESULTS: None of the dressings exhibited significant temperature increases nor produced significant distortion that influenced imaging quality. CONCLUSION: Our data suggests silver containing wound dressings do not cause a significant increase in dressing temperature or image distortion and thus their removal is not warranted for clinical MRI examinations. PMID- 19560872 TI - Action of estrogen on survival of basal forebrain cholinergic neurons: promoting amelioration. AB - Extensive studies during the past two decades provide compelling evidence that the gonadal steroid, estrogen, has the potential to affect the viability of basal forebrain cholinergic neurons. These observations reflect a unique ameliorative feature of estrogen as it restores and protects the cholinergic neurons against noxious stimuli or neurodegenerative processes. Hence, we first address the ameliorative function of estrogen on basal forebrain cholinergic neurons such as the actions of estrogen on neuronal plasticity of cholinergic neurons, estrogen induced memory enhancement and the ameliorative role of estrogen on cholinergic neuron related neurodegenerative processes such as Alzheimer's disease. Second, we survey recent data as to possible mechanisms underlying the ameliorative actions of estrogen; influencing the amyloid precursor protein processing, enhancement in neurotrophin receptor signaling and estrogen-induced non-classical actions on second messenger systems. In addition, clinical relevance, pitfalls and future aspects of estrogen therapy on basal forebrain cholinergic neurons will be discussed. PMID- 19560873 TI - Longitudinal changes in drug use severity and physical health-related quality of life among untreated stimulant users. AB - The primary objective of this study was to investigate whether drug use severity is associated with physical health-related quality of life (HRQL) over time. Data are from a longitudinal, multi-state, natural history community study of users of cocaine and/or methamphetamine who were interviewed at 6-month intervals over 2 years with a 79% follow-up participation rate. Physical HRQL was assessed with the physical component summary (PCS) of the SF-8 Health Survey and drug, alcohol, and psychiatric severity were all assessed with the Addiction Severity Index (ASI). Random coefficient regression analyses were conducted to test for longitudinal associations between the independent variables and SF-8 PCS scores. Reductions in drug use severity over time were accompanied by only minor improvements in SF-8 PCS scores, underscoring the potential long-term harm of illicit drug use on physical health. Greater psychiatric severity was strongly associated with lower SF-8 PCS scores, suggesting that clinical attention to mental health issues could potentially lead to improvements in perceived physical health as well as among stimulant users. PMID- 19560874 TI - Headache associated with visceral disorders is "parasympathetic referred pain". AB - Referred pain from visceral organs tends to be expressed on the specific area of body surface, called as Head's zone. Although it is well known that sympathetic referred pains of viscera appear on the body trunk, the fact that parasympathetic referred pains exist and are expressed on the head, sacrum and posterior thigh is not appreciated properly. Functional gastrointestinal diseases accompany frequently headache, and cyclic vomiting and recurrent abdominal pains in childhood progress to migraine later. Such clinical observations on relationship between headache and viscera suggest that longstanding disease processes of viscera could induce central sensitization of trigeminocervical nuclear complex, and express "parasympathetic referred pain" on the head, like sympathetic referred pain on the body trunk, that is headache. PMID- 19560875 TI - Hypothetical hormonal mechanism by which potassium-rich diets benefit patients with rheumatoid arthritis. AB - Patients with rheumatoid arthritis (RA) have significantly lower salivary and serum potassium (K) concentration, reduced total body K, and lower dietary K intake than healthy subjects. There may also be a subtle impairment in the hypothalamic-pituitary-adrenal (HPA) axis in patients with RA with both a poor cortisol secretion response as well as a lower adrenocorticotropin hormone (ACTH) response in relation to involved inflammatory factors. Patients with RA also exhibit an impaired Na+, K+-ATPase (NKA) activity which might promote the pro inflammatory cytokine secretion seen in RA. I will use these facts to support the mechanism I propose. There are no qualitative differences between the effects of endogenous cortisol and exogenously applied synthetic glucocorticoids (GCs), which are widely used to treat RA. All effects are transmitted via the same receptor. The GC, cortisol, plays a role in normal K homeostasis and the reverse is also seen with higher K intake leading to higher cortisol secretion and biosynthesis. Results of a recent clinical trial showed elevated serum cortisol followed K supplementation. I suggest that this is what alleviated RA symptoms. I would like to suggest a "Cortisol-K" theory as a mechanism for De Coti-Marsh's proposed "K theory" while not precluding the possibility of eventual proof of a cure, possibly from effects of K inside cells other than the adrenal glands. PMID- 19560876 TI - Henna (Lawsonia inermis) might be used to prevent mycotic infection. PMID- 19560877 TI - Is it the time for metformin to take place in adjuvant treatment of Her-2 positive breast cancer? Teaching new tricks to old dogs. AB - Breast cancer is the most common malignancy diagnosed among women. According to the new molecular subclassification, basal like and Her-2 positive breast cancers have the worst outcome and these are the ones in which chemotherapy is a must as a part of adjuvant treatment. New treatment options that could be used as an adjuvant maintenance treatment are still being investigated. Insulin hormone is one of the reasons of breast cancer recurrence and death in breast cancer survivors. Targeting insulin as a therapeutic modality in breast cancer could be an option in the adjuvant treatment of breast cancer. It seems that insulin may signal to activate a cascade of proliferative and anti-apoptotic events in the cancer cell. Metformin, an oral anti-diabetic known for 50 years, may also have direct effects on cancer cells. Metformin causes Her-2 suppression via the inhibition of mTOR in breast cancer cells. Thus, we believe that the time has arrived both to target insulin reduction and to alter Her-2 oncogene based molecular pathogenetic steps in breast cancer by using metformin as an adjuvant therapy in breast cancer patients. PMID- 19560878 TI - Self-reinforcing loop mechanism in epilepsy. AB - The evidence for a mutual dependence between convulsive activity and duration of the paradoxical sleep (PS) can be found from the literature. Convulsions diminish the PS time, on the other hand, PS restriction increases the sensitivity to convulsion-inducing stimuli. In epilepsy, the operation of this positive feedback loop mechanism is possibly promoted by elevated levels of enkephalin and norepinephrine that are characteristic of epileptics. The above supports the convulsions-PS interplay representing a relevant therapeutic target in epilepsy. In this connection, usage of PS duration increasing hypnotics like thalidomide, in combination with enkephalin and norepinephrine antagonists is thought to be of special importance. PMID- 19560879 TI - Anti-protein aggregation is a potential target for preventing delayed neuronal death after transient ischemia. AB - Brain ischemia has been an important risk factor for human being health, there is no effective medicine can be used to protect delayed neuronal injury or death secondary to blood reperfusion following ischemia. Recent discovery shows protein aggregation is an important factor resulting in ischemia-induced neuron death. Therefore, we propose the hypothesis that inhibiting protein aggregation may be an effective way to prevent delayed neuronal death after transient ischemia. At present, in vitro studies show some chemicals such as 4PBA (sodium 4 phenylbutyrate) and trehalose have the features of antagonizing protein aggregation in vitro. Moreover, polyQ-binding peptide (QBP1), geldanamycin, amino acids and amino acid derivatives have been also used in vitro to decrease aggregation and to increase protein stability. Although in vivo and systematical study should be performed to evaluate their effects of anti-protein aggregation, this enlightening us on using them to protect ischemic-induced neuronal death, and find new potential chemicals or methods which could be effective in keeping protein stable and prevent forming aggregates. PMID- 19560880 TI - A comparison of volumetric modulated arc therapy and conventional intensity modulated radiotherapy for frontal and temporal high-grade gliomas. AB - PURPOSE: Volumetric modulated arc therapy (VMAT), the predecessor to Varian's RapidArc, is a novel extension of intensity-modulated radiotherapy (IMRT) wherein the dose is delivered in a single gantry rotation while the multileaf collimator leaves are in motion. Leaf positions and the weights of field samples along the arc are directly optimized, and a variable dose rate is used. This planning study compared seven-field coplanar IMRT (cIMRT) with VMAT for high-grade gliomas that had planning target volumes (PTVs) overlapping organs at risk (OARs). METHODS AND MATERIALS: 10 previously treated patients were replanned to 60 Gy in 30 fractions with cIMRT and VMAT using the following planning objectives: 98% of PTV covered by 95% isodose without violating OAR and hotspot dose constraints. Mean OAR doses were maximally decreased without reducing PTV coverage or violating hotspot constraints. We compared dose-volume histogram data, monitor units, and treatment times. RESULTS: There was equivalent PTV coverage, homogeneity, and conformality. VMAT significantly reduced maximum and mean retinal, lens, and contralateral optic nerve doses compared with IMRT (p < 0.05). Brainstem, chiasm, and ipsilateral optic nerve doses were similar. For 2-Gy fractions, mean monitor units were as follows: cIMRT = 789 +/- 112 and VMAT = 363 +/- 45 (relative reduction 54%, p = 0.002), and mean treatment times (min) were as follows: cIMRT = 5.1 +/- 0.4 and VMAT = 1.8 +/- 0.1 (relative reduction 65%, p = 0.002). CONCLUSIONS: Compared with cIMRT, VMAT achieved equal or better PTV coverage and OAR sparing while using fewer monitor units and less time to treat high-grade gliomas. PMID- 19560881 TI - Variations in target volume definition for postoperative radiotherapy in stage III non-small-cell lung cancer: analysis of an international contouring study. AB - PURPOSE: Postoperative radiotherapy (PORT) in patients with completely resected non-small-cell lung cancer with mediastinal involvement is controversial because of the failure of earlier trials to demonstrate a survival benefit. Improved techniques may reduce toxicity, but the treatment fields used in routine practice have not been well studied. We studied routine target volumes used by international experts and evaluated the impact of a contouring protocol developed for a new prospective study, the Lung Adjuvant Radiotherapy Trial (Lung ART). METHODS AND MATERIALS: Seventeen thoracic radiation oncologists were invited to contour their routine clinical target volumes (CTV) for 2 representative patients using a validated CD-ROM-based contouring program. Subsequently, the Lung ART study protocol was provided, and both cases were contoured again. Variations in target volumes and their dosimetric impact were analyzed. RESULTS: Routine CTVs were received for each case from 10 clinicians, whereas six provided both routine and protocol CTVs for each case. Routine CTVs varied up to threefold between clinicians, but use of the Lung ART protocol significantly decreased variations. Routine CTVs in a postlobectomy patient resulted in V(20) values ranging from 12.7% to 54.0%, and Lung ART protocol CTVs resulted in values of 20.6% to 29.2%. Similar results were seen for other toxicity parameters and in the postpneumectomy patient. With the exception of upper paratracheal nodes, protocol contouring improved coverage of the required nodal stations. CONCLUSION: Even among experts, significant interclinician variations are observed in PORT fields. Inasmuch as contouring variations can confound the interpretation of PORT results, mandatory quality assurance procedures have been incorporated into the current Lung ART study. PMID- 19560882 TI - High-dose-rate intraoperative radiation therapy for recurrent head-and-neck cancer. AB - PURPOSE: To report the use of high-dose-rate intraoperative radiation therapy (HDR-IORT) for recurrent head-and-neck cancer (HNC) at a single institution. METHODS AND MATERIALS: Between July 1998 and February 2007, 34 patients with recurrent HNC received 38 HDR-IORT treatments using a Harrison-Anderson-Mick applicator with Iridium-192. A single fraction (median, 15 Gy; range, 10-20 Gy) was delivered intraoperatively after surgical resection to the region considered at risk for close or positive margins. In all patients, the target region was previously treated with external beam radiation therapy (median dose, 63 Gy; range, 24-74 Gy). The 1- and 2-year estimates for in-field local progression-free survival (LPFS), locoregional progression-free survival (LRPFS), distant metastases-free survival (DMFS), and overall survival (OS) were calculated. RESULTS: With a median follow-up for surviving patients of 23 months (range, 6-54 months), 8 patients (24%) are alive and without evidence of disease. The 1- and 2 year LPFS rates are 66% and 56%, respectively, with 13 (34%) in-field recurrences. The 1- and 2-year DMFS rates are 81% and 62%, respectively, with 10 patients (29%) developing distant failure. The 1- and 2-year OS rates are 73% and 55%, respectively, with a median time to OS of 24 months. Severe complications included cellulitis (5 patients), fistula or wound complications (3 patients), osteoradionecrosis (1 patient), and radiation-induced trigeminal neuralgia (1 patient). CONCLUSIONS: HDR-IORT has shown encouraging local control outcomes in patients with recurrent HNC with acceptable rates of treatment-related morbidity. Longer follow-up with a larger cohort of patients is needed to fully assess the benefit of this procedure. PMID- 19560884 TI - Early expansion of the intracranial CSF volume after palliative whole-brain radiotherapy: results of a longitudinal CT segmentation analysis. AB - PURPOSE: To assess cerebral atrophy after radiotherapy, we measured intracranial cerebrospinal fluid volume (ICSFV) over time after whole-brain radiotherapy (WBRT) and compared it with published normal-population data. METHODS AND MATERIALS: We identified 9 patients receiving a single course of WBRT (30 Gy in 10 fractions over 2 weeks) for ipsilateral brain metastases with at least 3 years of computed tomography follow-up. Segmentation analysis was confined to the tumor free hemi-cranium. The technique was semiautomated by use of thresholds based on scanned image intensity. The ICSFV percentage (ratio of ICSFV to brain volume) was used for modeling purposes. Published normal-population ICSFV percentages as a function of age were used as a control. A repeated-measures model with cross sectional (between individuals) and longitudinal (within individuals) quadratic components was fitted to the collected data. The influence of clinical factors including the use of subependymal plate shielding was studied. RESULTS: The median imaging follow-up was 6.25 years. There was an immediate increase (p < 0.0001) in ICSFV percentage, which decelerated over time. The clinical factors studied had no significant effect on the model. CONCLUSIONS: WBRT immediately accelerates the rate of brain atrophy. This longitudinal study in patients with brain metastases provides a baseline against which the potential benefits of more localized radiotherapeutic techniques such as radiosurgery may be compared. PMID- 19560883 TI - Radiosensitization of chemotherapy-refractory, locally advanced or locally recurrent breast cancer with trastuzumab: a phase II trial. AB - PURPOSE: Trastuzumab (Herceptin), an anti-human epidermal growth factor receptor 2 (HER2) antibody, has been shown to be an effective radiosensitizer in preclinical studies. The present Phase II trial evaluated trastuzumab plus radiotherapy in patients with HER2-positive, chemotherapy-refractory, locally advanced or locoregionally recurrent breast cancer. METHODS AND MATERIALS: Eligible patients had measurable disease, normal cardiac function, and biopsy confirmed residual HER2-positive disease. Patients received weekly trastuzumab (2 mg/kg intravenously), concurrent with radiotherapy (50 Gy) to the breast and regional lymph nodes for 5 weeks. If feasible, surgery followed radiotherapy. The primary endpoint was safety, and the secondary endpoint was efficacy (pathologic response and interval to symptomatic local progression). RESULTS: Of the 19 patients enrolled, 7 were ineligible and received radiotherapy alone and 12 received therapy per protocol. Of these 12 patients, 11 had a Stage T4 diagnosis. Grade 3 toxicities included skin (n = 2) and lymphopenia (n = 1). One patient experienced delayed wound healing after surgery. No patients developed symptomatic cardiac dysfunction. Of the 7 patients who had undergone mastectomy, 3 (43%) had a substantial pathologic response (complete response or microscopic residual disease), significantly more than a comparison cohort (2 of 38 or 5%, p = .02). The median interval to symptomatic local progression was not reached. The median overall survival was 39 months. CONCLUSION: This is the first prospective trial providing evidence for a radiosensitizing effect of trastuzumab in breast cancer. The combination of trastuzumab and radiotherapy was well tolerated. PMID- 19560886 TI - Detection and characterization of porcine sapoviruses from asymptomatic animals in Irish farms. AB - Caliciviruses are an important cause of gastroenteritis in humans and animals. Molecular analysis of the polymerase and capsid genes of porcine caliciviruses, sapoviruses (SaVs) and noroviruses (NoVs), has demonstrated a broad range of genetic diversity but information on their epidemiology and pathogenic role in pigs is limited. In this study, 292 faecal samples were obtained from 4-5 to 8-9 week old asymptomatic pigs from four porcine herds in Ireland during 2005-2007 and were screened by RT-PCR using calicivirus-specific primers. Only seven samples from two porcine herds tested positive for porcine calicivirus. By sequence analysis of the partial RNA-dependent RNA polymerase (RdRp) fragment, six samples from one such herd were closely related to each other (>98% nucleotide identity) and were characterised as genogroup (GG) III (Cowden-like) porcine SaVs. These viruses demonstrated an amino acid (aa) identity of 81.3 98.6% to GGIII SaVs. Conversely, one calicivirus strain, 9/07/Ire (identified from a different herd in 2007), was distantly related to GIII SaVs and displayed 94.6-98.6% aa identity to rare K7-like porcine caliciviruses, representatives of a potential novel SaV genogroup (GGVII), described previously in Japan and the USA. Circulation of SaVs in asymptomatic animals might be a mechanism of virus persistence in porcine populations and should be considered with respect to understanding the epidemiology of these viruses in porcine herds. PMID- 19560885 TI - Chronic alcohol consumption from adolescence-to-adulthood in mice--effect on growth and social behavior. AB - Experimentation with alcohol is common during adolescence. However the long-term consequences from moderate alcohol use during adolescence development are not clear. Using a two-bottle free-choice paradigm in the home-cage setting, we studied adolescent mice (4 weeks old) across a 6-week time span of the adolescence-to-adulthood development period. Adolescent mice readily reached a steady level of alcohol consumption and maintained this level throughout the 6 week period. Chronic alcohol consumption resulted in reduced growth in adolescent mice, as well as accelerated acclimation to a novel environment. During a social interaction test, similar levels of initial social investigation and subsequent habituation were observed in both the chronic alcohol and the water-only control groups. However, chronic alcohol self-administration resulted in impaired social recognition and decreased social play/fight behavior. Taken together, these results indicated that chronic alcohol consumption across adolescence development negatively impacted both physical growth and social behavior in mice, highlighting the detrimental consequences from prolonged alcohol drinking in adolescence. PMID- 19560887 TI - [Differentiating bacterial from viral meningitis: contribution of nonmicrobiological laboratory tests]. AB - In most cases, differentiating viral from bacterial meningitis is relatively easy, based on clinical examination, CSF appearance and results of CSF examination (cytology, biochemistry and Gram stain). However, in about 20% of cases, this diagnosis may be difficult. For such cases, additional non microbiological tests may be helpful. CSF lactate level is a good predictor of bacterial meningitis for values greater than 3.5 mmol/l. Serum procalcitonin is effective to discriminate between bacterial and viral meningitis, using a threshold between 1 and 2 ng/ml, although this parameter may fail in individual situations. Accurate diagnosis scores or models have been validated and may be used in routine clinical practice, especially in emergency rooms, both for adults and children to help identify patients with a very low probability of bacterial meningitis in whom antibiotic may thus be avoided. PMID- 19560888 TI - [Treatment of hepatitis B: new perspectives]. AB - Despite the development of new anitiviral agents, the treatment of chronic hepatitis B remains a major clinical challenge. Major achievements have been made with the rationale use of antivirals exhibiting a complementary cross resistance profile to prevent antiviral drug resistance. The current concept of modern antiviral therapy of chronic hepatitis B relies on a precise virologic monitoring and early treatment adaptation to prevent drug resistance. The difficulty of achieving viral clearance and the risk of drug resistance development are major arguments to continue research in the field of antivirals and to identify new targets for therapy. The development of true combination therapy is highly desirable to fulfil the objective of long-term viral suppression, clearance of viral cccDNA and infected cells and ultimately cure of the disease. PMID- 19560889 TI - [Liver fibrosis: from pathophysiology to therapeutic openings]. AB - Understanding of liver fibrosis pathogenesis has undergone tremendous advances over the past twenty years. In this respect, demonstration of the reversibility of fibrosis was a major turnpoint. The panel of therapeutic targets is continuously expanding. Clinical development has however remained limited, heretofore, but should rapidly progress owing to the availability of accurate non invasive methods for assessment of fibrosis, to improvement in the selection patients included in therapeutic trials, and to the development of cell specific targeting devices for agents at risk of adverse effects. PMID- 19560890 TI - Diffusion weighted imaging and estimation of prognosis using apparent diffusion coefficient measurements in ischemic stroke. AB - OBJECTIVE: Estimation of the prognosis of infarction by using diffusion weighted imaging (DWI) and quantitative apparent diffusion coefficient (ADC) measurements. METHODS: 23 patients having acute stroke symptoms with verified infarction in magnetic resonance imaging (MRI) were included in this study. Their MRI studies were performed between 6 and 12h after the onset of their symptoms and were repeated on the fifth day. The infarction volumes were calculated by using DWI and the patients were divided into two groups as the ones having an expansion in the infarction area (group 1, n=16) and the others having no expansion in the infarction area (group 2, n=7). Quantitative ADC values were estimated. The groups were compared in terms of the ADC values on ADC maps obtained from DWI, performed during the between 6 and 12h from the onset of the symptoms, referring to the core of the infarction (ADC(IC)), ischemic penumbra (ADC(P)) and the nonischemic parenchymal tissue (ADC(N)). P values<0.05 were accepted to be statistically significant. RESULTS: During the between 6 and 12h mean infarction volume calculated by DWI was 23.3cm(3) for group 1 patients (ranging from 1.1 to 68.6) and this was found to be 40.3cm(3) (ranging from 1.8 to 91.5) on the fifth day. For the group 2 patients these values were found to be 42.1cm(3) (ranging from 1 to 94.7) and 41.9 (ranging from 1 to 94.7) for the same intervals respectively. A significant statistical result was failed to be demonstrated between the mean ADC(IC) and ADC(N) values (p=0.350 and p=0.229 respectively). However the comparison of the ADC(P) values between the groups was found to be highly significant (p<0.001). When the differences between the ADC(P) and ADC(IC) and ADC(N) and ADC(P) were compared the results proved to be statistically significant (p=0.038 and p<0.001 respectively). CONCLUSIONS: We believe that ADC results that would be obtained from the core and the penumbra of the infarction area will be beneficial in the estimation of the infarction prognosis and in the planning of a treatment protocol. PMID- 19560892 TI - Identification and characterization of potential impurities of valsartan, AT1 receptor antagonist. AB - Five impurities (related substances) were detected during the impurity profile study of an antihypertensive drug substance, valsartan. A simple gradient high performance liquid chromatographic method (HPLC) and liquid chromatography-mass spectrometry (LC-MS) were used for the detection. Based on the spectral data (IR, NMR and MS) followed by synthesis, these impurities were characterized as (S)-N (1-carboxy-2-methylprop-1-yl)-N-[2'-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]amine (impurity I); (S)-N-(1-carboxy-2-methylprop-1-yl)-N-(5-phenylthio)pentanoyl-N-[2' (1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]amine (impurity II); (S)-N-(1-carboxy-2 methylprop-1-yl)-N-(5-phenyl)pentanoyl-N-[2'-(1H-tetrazol-5-yl)-biphenyl-4 ylmethyl]amine (impurity III); (S)-N-(1-carboxy-2-methylprop-1-yl)-N-4-pentenoyl N-[2'-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]amine (impurity IV); (S)-N-(1 carboxy-2-methylprop-1-yl)-N-(5-hydroxy)pentanoyl-N-[2'-(1H-tetrazol-5-yl) biphenyl-4-ylmethyl]amine (impurity V). PMID- 19560891 TI - Community structure of microorganisms associated with reddish-brown iron-rich snow. AB - Reddish-brown colored snow, containing spherical brown particles, has been observed in several mires in Japan. In order to characterize this remarkable phenomenon, the microbial community and chemical species in snow were analyzed. A core sample of snow which had a colored region was investigated and it revealed vertical shifts in physicochemical characteristics and the microbial community structure. The abundance of particles peaked within the colored layer, and correlated with the amount of reducible Fe(III). The interstitial water of the colored layer was enriched with Fe(II), and characterized by reduced concentration of dissolved methane. The bacterial community in the colored region was characterized by higher relative abundance of iron-reducing bacteria and methanotrophs. Aggregates of the brown particles were found as precipitates in snow melt pools, and were subjected to cloning analyses targeting several different genes. The majority of bacterial 16S rRNA gene clones belonged to the class Betaproteobacteria or the phylum Bacteroidetes. No snow algae were detected in the eukaryotic small subunit rRNA gene clone library. As a possible carbon source to sustain the community in the snow, involvements of carbon dioxide and methane were investigated by analyzing the genes involved in their assimilation. In the analyses of genes for ribulose-1,5-biphosphate carboxylase/oxygenase, clones related to sulfur oxidizers were obtained. The analysis of particulate methane monooxygenase genes indicated dominance of Methylobacter species. These results emphasized the uniqueness of this phenomenon, and iron reducers of the genus Geobacter are suggested to be the key organisms that could be investigated in order to understand the mechanism of this phenomenon. PMID- 19560893 TI - Patients' understanding of the concepts of health and quality of life. AB - OBJECTIVE: The aim of this study was to identify how persons' with rheumatoid arthritis (RA) understand the concepts of health and quality of life (QoL). METHODS: A phenomenograpic approach was used to gauge variations in understanding of health and QoL. Semistructured interviews were conducted with 22 persons having RA. The interviews were audiotaped, transcribed verbatim, and analyzed, resulting in a descriptive system consisting of categories and subcategories. RESULTS: Health was primarily associated with being healthy/being free from disease, being able to function normally, experiencing well-being, and having a healthy lifestyle. Two above-mentioned domains, "being healthy" and "being able to function normally" overlapped with respondents' understanding of QoL. Additional perceived domains included attitudes towards life and, social and physical environments. CONCLUSION: The results show that patients' understanding of the two concepts of health and QoL partially overlap and partially differ. PRACTICE IMPLICATIONS: Patients' understanding of the concepts is individual and diverse, which needs to be taken into account in patient-physician consultations to ensure good communication. Furthermore, the interpretation of results based on patient-reported outcomes instruments measuring health status and/or QoL needs to be further studied over time to identify possible changes in these conceptions. PMID- 19560894 TI - Limits to truth-telling: neurologists' communication in conversion disorder. AB - OBJECTIVE: Neurologists face a dilemma when communicating with their conversion disorder patients - whether to be frank, and risk losing the patient's trust, or to disclose less, in the hope of building a therapeutic relationship. This study reports how neurologists in the UK described dealing with this dilemma in their practice. METHODS: Practicing consultant neurologists from an NHS region were recruited by snowball sampling. Twenty-two of 35 consultants in the region were interviewed in depth, and the interviews qualitatively analysed. RESULTS: The neurologists were reluctant to disclose conversion disorder as a differential diagnosis until they were certain. They were guided by the receptivity of their patients as to how psychological to make their eventual explanations, but they did not discuss their suspicions about feigning. They described their communications as much easier now than they had seen in training. CONCLUSION: Neurologists adapt their disclosure to their patients, which facilitates communication, but imposes some limits on truth-telling. In particular, it may sometimes result in a changed diagnosis. PRACTICE IMPLICATIONS: An optimum strategy for communicating diagnoses will need to balance ethical considerations with demonstrated therapeutic benefit. PMID- 19560895 TI - Goal-setting for behavior change in primary care: an exploration and status report. AB - OBJECTIVE: This paper explores the behavior change method of goal-setting and reviews the literature on goal-setting in primary care for patients with chronic conditions. METHODS: A literature search was conducted resulting in eight articles meeting the criteria of goal-setting interventions in primary care for adults or adolescents with chronic conditions. RESULTS: Hypotheses are advanced that goal-setting is generally conducted by collaboratively working with patients to set short-term and specific goals, with follow-up to provide feedback to patients. The articles reviewed generally confirmed these hypotheses. This review did not focus on clinical outcomes, but on the processes of engaging patients in goal-setting discussions. CONCLUSION: Evidence that goal-setting is superior to other behavior change methods has not been shown. Since goal-setting is being utilized as a behavior change technique in many primary care sites, primary care practices can benefit from information on how best to implement this innovation. PRACTICE IMPLICATIONS: Generally, clinicians are minimally involved in goal setting discussions with their patients. Engaging patients in goal-setting can be done with interactive computer programs and non-clinical members of the primary care team. PMID- 19560897 TI - [Liver transplantation: 20 years of progress]. PMID- 19560896 TI - [Renal failure following liver transplantation]. AB - The cumulative incidence of chronic kidney disease after liver transplantation (LT) is on the order of 40 to 50% at 1 year and over 50% at 5 years, and that of pre-end-stage renal failure 5-10% at 1 year and 10-20% at 5 and 10 years. Several variables appear to be independently associated with onset of kidney failure: age, sex (male), ethnicity (non-Asian), low glomerular filtration rate (GFR) before transplantation, use of renal replacement therapy before LT, diabetes before LT, HCV carriage, postoperative onset of acute renal failure, the year of transplantation (before or after 1994). Various factors cause chronic kidney disease after LT. Calcineurin inhibitors, specifically cyclosporine and tacrolimus, but also diabetes, nephroangiosclerosis, previous use of hydroxyethylstarch, play a major role in the onset of postgraft kidney failure. It is generally agreed that the nephrotoxicity of calcineurin inhibitors is in part dose-dependent and that a reduction in the dose can improve renal function. Nonetheless, the lesions are in large part irreversible. Trials are required to test interventions early after the LT, as soon as the first signs of kidney failure appear. Moreover, although the effect is dose-dependent, the relation with blood concentration of the drug is very imperfect, so any intervention must reduce the dose and not just the concentration to improve renal function. The introduction of new immunosuppressive drugs that are not nephrotoxic, such as mycophenolate mofetil, mTOR inhibitors, and anti-CD25 monoclonal antibodies [basiliximab and daclizumab (withdrawn from the market)], allow primary or secondary prevention of nephrotoxicity, with a partial or complete reduction in calcineurin inhibitors. Other interventions useful to limiting kidney failure after LT are the correction of hypertension, diabetes, and hyperlipidemia. PMID- 19560898 TI - Subject-independent mental state classification in single trials. AB - Current state-of-the-art in Brain Computer Interfacing (BCI) involves tuning classifiers to subject-specific training data acquired from calibration sessions prior to functional BCI use. Using a large database of EEG recordings from 45 subjects, who took part in movement imagination task experiments, we construct an ensemble of classifiers derived from subject-specific temporal and spatial filters. The ensemble is then sparsified using quadratic regression with l(1) regularization such that the final classifier generalizes reliably to data of subjects not included in the ensemble. Our offline results indicate that BCI naive users could start real-time BCI use without any prior calibration at only very limited loss of performance. PMID- 19560899 TI - The impact of Le Fort I advancement and bilateral sagittal split osteotomy setback on ventilation during sleep. AB - The impact of orthognathic surgery for class III malocclusion on ventilation during sleep was examined using a comparison of pre- and post-surgical respiratory parameters. 21 patients with both maxillary hypoplasia and mandibular excess underwent Le Fort I osteotomy and advancement together with bilateral sagittal split osteotomy (BSSO) setback. Respiratory parameters, ECG and position of the body were monitored before surgery and postoperatively after the fixation removal (mean 8.5 months). Average Le Fort I advancement was 4.44 mm, BSSO setback was 4.96 mm. Generally, the orthognathic procedure worsened breathing function during sleep, as reflected in significant increase of index of flow limitations and decrease in oxygen saturation. The posterior airways space decreased to 75% of its original volume, the distance between mandibular plane and hyoid bone increased to 133%. The results indicate that bimaxillary surgery for class III malocclusion increased upper airway resistance. A young person would probably be able to balance such a decline in respiratory function using different adaptive mechanisms, but the potential impact of orthognathic surgery on the upper airways should be incorporated in a treatment plan. PMID- 19560900 TI - Assessing the impact of cannabis use on trends in diagnosed schizophrenia in the United Kingdom from 1996 to 2005. AB - A recent systematic review concluded that cannabis use increases risk of psychotic outcomes independently of confounding and transient intoxication effects. Furthermore, a model of the association between cannabis use and schizophrenia indicated that the incidence and prevalence of schizophrenia would increase from 1990 onwards. The model is based on three factors: a) increased relative risk of psychotic outcomes for frequent cannabis users compared to those who have never used cannabis between 1.8 and 3.1, b) a substantial rise in UK cannabis use from the mid-1970s and c) elevated risk of 20 years from first use of cannabis. This paper investigates whether this has occurred in the UK by examining trends in the annual prevalence and incidence of schizophrenia and psychoses, as measured by diagnosed cases from 1996 to 2005. Retrospective analysis of the General Practice Research Database (GPRD) was conducted for 183 practices in England, Wales, Scotland and Northern Ireland. The study cohort comprised almost 600,000 patients each year, representing approximately 2.3% of the UK population aged 16 to 44. Between 1996 and 2005 the incidence and prevalence of schizophrenia and psychoses were either stable or declining. Explanations other than a genuine stability or decline were considered, but appeared less plausible. In conclusion, this study did not find any evidence of increasing schizophrenia or psychoses in the general population from 1996 to 2005. PMID- 19560902 TI - Annual macrolide prescription rates and the emergence of macrolide resistance among Streptococcus pneumoniae in Canada from 1995 to 2005. AB - Over the last 20 years, Canadian pneumococcal surveillance studies have documented a steady rise in macrolide resistance. In the current study, we probed the nature of associations between the emergence of macrolide-resistant Streptococcus pneumoniae in Canada and changes in macrolide (azithromycin, clarithromycin and erythromycin) prescription rates. Macrolide susceptibility testing data for respiratory tract isolates of S. pneumoniae (n=15109) were acquired from two published national Canadian surveillance databases, and dispensed outpatient macrolide prescription data were acquired from the proprietary Intercontinental Medical Statistics (IMS) Health Canada CompuScript database. Nationally, macrolide resistance increased from 3.7% in 1995 to 19.0% in 2005 (P=0.003) as the annual macrolide prescription rate increased from 106.7 to 123.2 prescriptions/1000 persons per year (P=0.003). From 1995 to 2005, azithromycin and clarithromycin prescriptions increased from 4.8 to 52.5 prescriptions/1000 persons per year (P<0.0001) and from 24.7 to 58.4 prescriptions/1000 persons per year (P=0.005), respectively, whilst erythromycin prescriptions decreased from 77.2 to 12.3 prescriptions/1000 persons per year (P<0.0001). By univariate regression analysis, increasing rates of azithromycin (R(2)=0.931; P<0.0001) and clarithromycin (R(2)=0.725; P=0.0009) prescriptions and a decreasing rate of erythromycin prescriptions (R(2)=-0.963; P<0.0001) were all associated with increasing macrolide resistance from 1995 to 2005. Multivariate regression analysis showed that a model including all three macrolide prescription rates provided the best fit to the trend of increasing macrolide resistance. When the data were analysed by provincial origin, no statistically significant associations were found between prescription rates of any macrolide and macrolide resistance rates by univariate and multivariate regression analyses. We conclude that increasing macrolide resistance among respiratory isolates of pneumococci in Canada from 1995 to 2005 was associated both with decreasing prescriptions for erythromycin and concurrent increases in prescriptions for azithromycin and clarithromycin (azithromycin>clarithromycin by univariate regression analysis). Resistance development is complex and factors other than macrolide use may also be associated with observed increases in macrolide resistance in Canada from 1995 to 2005. PMID- 19560901 TI - Defining and predicting functional outcome in schizophrenia and schizophrenia spectrum disorders. AB - BACKGROUND: To assess criteria and to identify predictive factors for functional outcome. The criteria should cover all domains proposed by the Remission in Schizophrenia Working Group. METHOD: PANSS ratings were used to evaluate the symptomatic treatment outcome of 262 inpatients with schizophrenia spectrum disorders within a naturalistic multicenter trial. Functional remission was defined as a GAF score >61 (Global Assessment of Functioning Scale), SOFAS score >61 (Social and Occupational Functioning Scale) and a SF-36 mental health subscore >40 (Medical Outcomes Study-Short Form Health Survey). Multivariate logistic regression and CART analyses were used to determine valid clinical and sociodemographic predictors. RESULTS: In total, 52 patients (20%) fulfilled the criteria for functional remission, 125 patients (48%) achieved symptomatic resolution and when criteria for functional remission and symptomatic resolution were combined 33 patients (13%) achieved complete remission. Younger age, employment, a shorter duration of illness, a shorter length of current episode, less suicidality, and a lower PANSS negative and global subscore at admission were predictive of functional remission. The regression model showed a predictive value of more than 80%. CONCLUSIONS: A significant association was found between functional remission and symptomatic resolution, indicating reasonable validity of the proposed definition for functional outcome. The revealed predictors for functional treatment outcome emphasize the need for psychosocial and vocational rehabilitation in schizophrenic patients. PMID- 19560903 TI - Analysis of plasmid-mediated multidrug resistance in Escherichia coli and Klebsiella oxytoca isolates from clinical specimens in Japan. AB - This study investigated the relationship of plasmid-mediated quinolone resistance (PMQR) and aminoglycoside resistance among oxyimino-cephalosporin-resistant Escherichia coli (n=46) and Klebsiella oxytoca (n=28) clinical isolates in Japan. Seventy-three isolates appeared to produce an extended-spectrum beta-lactamase (ESBL) and one K. oxytoca isolate produced IMP-1 metallo-beta-lactamase (MBL). Polymerase chain reaction (PCR) and sequencing confirmed that eight CTX-M-9/SHV 12-producing isolates, one IMP-1-producing K. oxytoca isolate, and six ESBL positive E. coli isolates respectively possessed PMQR genes qnrA1, qnrB6, and aac(6')-Ib-cr. All qnr-positive isolates also carried either aac(6')-Ib or aac(6')-IIc aminoglycoside acetyltransferase genes. Resistance determinants to beta-lactams, quinolones and aminoglycosides were co-transferred with a plasmid of ca. 140 kb. The qnrA1 gene was located downstream of insertion sequence ISCR1 in complex class 1 integrons. A novel qnrA1-carrying class 1 integron with the cassette arrangement aac(6')-IIc-aadA2 as well as a unique class 1 integron with bla(IMP-1)-aac(6')-IIc cassettes on the plasmid carrying qnrB6 were found in K. oxytoca isolates. We describe the identification of qnrB6 and aac(6')-Ib-cr and the close association of qnr with aac(6')-Ib and aac(6')-IIc for the first time in clinical isolates producing ESBL or MBL in Japan. PMID- 19560904 TI - Klebsiellapneumoniae isolates possessing KPC beta-lactamase in Israel, Puerto Rico, Colombia and Greece. PMID- 19560905 TI - Neural response to lidocaine in healthy subjects. AB - Recent studies suggest that some of cocaine's central nervous system (CNS) effects may be mediated through its sodium channel inhibiting local anesthetic properties. Local anesthetics that lack cocaine's strong affinity for the dopamine transporter (DAT) also produce sensory and mood effects, further suggesting a role for this neural pathway. Due to an absence of affinity at the DAT, the local anesthetic lidocaine may offer the potential to assess sodium channel activity in vivo in humans. To assess the utility of lidocaine as a CNS probe, we determined regional cerebral blood flow (rCBF) with single photon emission computed tomography (SPECT) following the intravenous administration of lidocaine (0.5 mg/kg) and compared this response to procaine (0.5 mg/kg and 1.0 mg/kg), a local anesthetic with partial affinity for the DAT, and saline. Infusions were administered in nine healthy female controls over a 10-day period with at least 2 days between each scan. Increased rCBF was observed following lidocaine, relative to saline, in the insula, caudate, thalamus, and posterior cingulate. Decreased rCBF was detected in a different region of the posterior cingulate. In general, increases in rCBF were more marked following lidocaine relative to procaine. Mood and sensory changes following lidocaine were limited and significantly less than those induced by either dose of procaine. There were no significant changes in blood pressure or heart rate following either medication. These findings suggest that lidocaine can be safely used to assess sodium channel function in persons with addictive and other psychiatric disorders. PMID- 19560907 TI - Sexually dimorphic gray matter volume reduction in patients with panic disorder. AB - While clinical features of panic disorder show significant sexual dimorphism, previous structural MRI studies have not sufficiently controlled for sex when looking at regional brain abnormalities in panic disorder. Using optimized voxel based morphometry (VBM), regional gray matter volume was compared between 24 patients (male/female: 9/15) with panic disorder and 24 healthy subjects matched for age and sex. Significant gray matter volume reductions were found in the bilateral dorsomedial and right ventromedial prefrontal cortices, right amygdala, anterior cingulate cortex, bilateral insular cortex, occipitotemporal gyrus and left cerebellar vermis in the patients compared with the controls. Among these regions, the VBM revealed significant sexual dimorphism: volume reduction in the right amygdala and the bilateral insular cortex was significantly greater in the males, while reduction in the right superior temporal gyrus was greater in females. Furthermore, a significant reduction in the dorsolateral and ventrolateral prefrontal cortices, thalamus, and parietal cortex was specific to the female patients. The present study demonstrated the morphological changes in extensive brain regions of patients with panic disorder compared with the sex matched controls. The current results further suggested that the sexually dimorphic clinical phenotypes of panic disorder might have a neurobiological background even at the structural level of the brain. PMID- 19560906 TI - Brain responses to repeated visual experience among low and high sensation seekers: role of boredom susceptibility. AB - To better understand individual differences in sensation seeking and its components, including boredom susceptibility and experience seeking, we examined brain responses of high and low sensation seekers during repeated visual experience. Individuals scoring in the top and bottom quartiles from a college aged population on the Brief Sensation-Seeking Scale (BSSS) participated in an event-related potentials (ERPs) experiment. Line drawings of common objects were randomly intermixed and presented 1-3 times. Sixty-four channel ERP responses were recorded while participants classified items as "man-made" or "not man-made" in a repetition priming task. The two groups showed different ERP responses at frontal electrode sites after seeing a visual stimulus for 400-800 ms. The frontal late positive components (LPC) showed different habituation of ERP responses to new and studied repeated objects between high and low sensation seekers. Source localization analysis (LORETA) indicated that during visual stimulus adaptation the ventral pre-frontal cortex showed lack of frontal involvement among high sensation seekers. Furthermore, frontal LPC latencies during repeated visual exposure correlated with boredom susceptibility and experience seeking subscales. The distinct profiles of brain responses to repeated visual experience in high and low sensation seekers provide evidence that individual differences in neural adaptation can be linked to personality dimensions. PMID- 19560908 TI - Geographical variation in radiotherapy services across the UK in 2007 and the effect of deprivation. AB - AIMS: Modelling of demand has shown substantial underprovision of radiotherapy in the UK. We used national audit data to study geographical differences in radiotherapy waiting times, access and dose fractionation across the four countries of the UK and between English strategic health authorities. MATERIALS AND METHODS: We used a web-based tool to collect data on diagnosis, dose fractionation and waiting times on all National Health Service patients in the UK starting a course of radiotherapy in the week commencing 24 September 2007. Cancer incidence for the four countries of the UK and for England by primary care trust was used to model demand for radiotherapy aggregated by country and by strategic health authority. RESULTS: Across the UK, excluding skin cancer, 2504 patients were prescribed 33 454 fractions in the audit week. Waits for radical radiotherapy exceeded the recommended 4 week maximum for 31% of patients (range 0 62%). Fractions per million per year ranged from 17 678 to 36 426 and radical fractions per incident cancer ranged from 3.0 to 6.7. Patients who were treated received similar treatment in terms of fractions per radical course of radiotherapy (18.2-23.0). Access rates ranged from 25.2 to 48.8%, nearing the modelled optimum of 50.7% in three regions. Fractions per million prescribed as a first course of treatment varied from 43.9 to 90.3% of modelled demand. The percentage of patients failing to meet the 4 week Joint Council for Clinical Oncology target for radical radiotherapy rose as activity rates increased (r=0.834), indicating a mismatch of demand and capacity. In England, a comparison between strategic health authorities showed that increasing deprivation was correlated with lower rates of access to radiotherapy (r=-0.820). CONCLUSIONS: There are substantial differences across the UK in the radiotherapy provided to patients and its timeliness. Radiotherapy capacity does not reflect regional variations in cancer incidence across the UK (3618-5800 cases per million per year). In addition, deprivation is a major unrecognised influence on radiotherapy access rates. In regions with higher levels of deprivation, fewer patients with cancer receive radiotherapy and the proportion treated radically is lower. This probably reflects late presentation with advanced disease, poor performance status and co-morbid illness. To provide an equitable, evidence-based service, the needs of the local population should be assessed using demand modelling based on local cancer incidence. Ideally this should include data on deprivation, performance status and stage at presentation. The results should be compared with local radiotherapy activity data to understand waits, access and dose fractionation in order to plan adequate provision for the future. The development of a mandatory radiotherapy data set in England will facilitate this, but to assist change it is essential that the results are analysed and fed back to clinicians and commissioners. PMID- 19560909 TI - In vitro micronucleus assay for cigarette smoke using a whole smoke exposure system: a comparison of smoking regimens. AB - Previous studies on the biological assessment of cigarette smoke (CS) mainly focused on the total particulate matter (TPM) collected using a Cambridge filter or gas vapor phase (GVP) bubbled through phosphate-buffered saline (PBS). To study the effects of native CS in vitro, direct exposure methods have been developed. Meanwhile, in vitro micronucleus (MN) assays have been reported to evaluate the mutagenicity of CS. The objective of this research is to investigate the MN-inducing activity of whole smoke (WS) and GVP using a whole smoke exposure system, CULTEX((R)), which allows direct exposure of cultured cells to native CS at the air/liquid interface (ALI). CS was generated according to the International Organization for Standardization (ISO; 35ml, 2s, once per 60s) or the Health Canada Intensive (HCI; 55ml, 2s, once per 30s, with complete ventilation block) regimens and Chinese hamster lung (CHL/IU) cells were then exposed to this smoke. Dosages were adjusted according to the amount of smoke entering the actual exposure position. Under both smoking regimens, WS and GVP from 2R4F reference cigarettes induced MN responses. The concept of the dosage and similar dose-response relationships between theoretical and monitored dosage values under the two regimens enabled us to compare the MN-inducing activities of cigarettes in the direct exposure assay, even in the case of various experimental settings or different TPM amounts. MN-inducing activities of 2R4F under the ISO regimen seemed to be higher than those under HCI estimated by the TPM equivalent calculated values. PMID- 19560910 TI - Increased biological hydrogen production by deletion of hydrogen-uptake system in photosynthetic bacteria. AB - Hydrogenases are the key enzymes for the biological hydrogen production, which can be classified as H(2)-uptake hydrogenase and H(2)-production hydrogenase. The genes encoding a membrane-bound [NiFe]-hydrogenase (MBH), which is mainly responsible for hydrogen uptake, from the photosynthetic bacterium Allochromatium vinosum was cloned and sequenced. It consist of two structural genes (hydS, hydL) and two intergenic genes (isp1, isp2), which are therefore organized as hydS-isp1 isp2-hydL. This is different from the arrangement of other typical hydrogenase gene clusters. A deletion mutant-strain PhihydSL, lacking isp1, isp2, partial hydS and hydL genes, was constructed by marker-exchange mutagenesis. Under dark fermentative conditions, the hydrogen production yield by this mutant increased by 62%. The result suggests that the disruption of MBH could greatly improve the hydrogen production in the cells by decreasing the hydrogen uptake. PMID- 19560912 TI - Multi-criteria decision analysis for waste management in Saharawi refugee camps. AB - The aim of this paper is to compare different waste management solutions in Saharawi refugee camps (Algeria) and to test the feasibility of a decision-making method developed to be applied in particular conditions in which environmental and social aspects must be considered. It is based on multi criteria analysis, and in particular on the analytic hierarchy process (AHP), a mathematical technique for multi-criteria decision making (Saaty, T.L., 1980. The Analytic Hierarchy Process. McGraw-Hill, New York, USA; Saaty, T.L., 1990. How to Make a Decision: The Analytic Hierarchy Process. European Journal of Operational Research; Saaty, T.L., 1994. Decision Making for Leaders: The Analytic Hierarchy Process in a Complex World. RWS Publications, Pittsburgh, PA), and on participatory approach, focusing on local community's concerns. The research compares four different waste collection and management alternatives: waste collection by using three tipper trucks, disposal and burning in an open area; waste collection by using seven dumpers and disposal in a landfill; waste collection by using seven dumpers and three tipper trucks and disposal in a landfill; waste collection by using three tipper trucks and disposal in a landfill. The results show that the second and the third solutions provide better scenarios for waste management. Furthermore, the discussion of the results points out the multidisciplinarity of the approach, and the equilibrium between social, environmental and technical impacts. This is a very important aspect in a humanitarian and environmental project, confirming the appropriateness of the chosen method. PMID- 19560911 TI - Cell biology of the ESCRT machinery. AB - The ESCRT (endosomal sorting complex required for transport) machinery comprises a set of protein complexes that regulate sorting and trafficking into multivesicular bodies en route to the lysosome. The physical mechanism responsible for generating lumenal vesicles in this pathway is unknown. Here we review recent studies suggesting that components of the ESCRT-III complex drive lumenal vesicle formation and consider possible mechanisms for this reaction. PMID- 19560913 TI - Robotic and laparoscopic surgery: cost and training. AB - Robotic prostatectomy training as part of mainstream surgical training will be difficult. The primary problems revolve around the inconsistencies of standard sugery. Many surgeons are still in the learning curve, as is the understanding of the true capabilities of the robot. The important elements of robotic surgery actually enhance basic laparoscopic techniques. The prostate has been shown to be an organ where this new technology has a niche. As we move toward cross specialty use the robot although extremely expensive, may be the best way to train the laparoscopic surgeon of the future. PMID- 19560914 TI - Using porphyritic andesite as a new additive for improving hydrolysis and acidogenesis of solid organic wastes. AB - The effects of porphyritic andesite on the hydrolysis and acidogenesis of solid organic wastes were investigated by batch and continuous experiments using a rotational drum fermentation system. The results of the batch experiment show that if porphyritic andesite (1%, 3%, and 5% reactants) is added initially, the pH level increases and hydrolysis and acidogenesis are accelerated. The highest surface based hydrolysis constant (26.4x10(-3) kgm(-2) d(-1)) and volatile solid degradation ratio (43.3%) were obtained at a 1% porphyritic andesite addition. In the continuous experiment, porphyritic andesite elevated the first order hydrolysis constant from 13.10x10(-3) d(-1) to 18.82x10(-3) d(-1). A particle mean diameter reduction rate of 33.05 microm/d and a volatile solid degradation rate of 3.53 g/L d(-1) were obtained under the hydraulic retention time of 4, 8, 12 and 16 d. PMID- 19560915 TI - Bio-oil production from fast pyrolysis of waste furniture sawdust in a fluidized bed. AB - The amount of waste furniture generated in Korea was over 2.4 million tons in the past 3 years, which can be used for renewable energy or fuel feedstock production. Fast pyrolysis is available for thermo-chemical conversion of the waste wood mostly into bio-oil. In this work, fast pyrolysis of waste furniture sawdust was investigated under various reaction conditions (pyrolysis temperature, particle size, feed rate and flow rate of fluidizing medium) in a fluidized-bed reactor. The optimal pyrolysis temperature for increased yields of bio-oil was 450 degrees C. Excessively smaller or larger feed size negatively affected the production of bio-oil. Higher flow and feeding rates were more effective for the production of bio-oil, but did not greatly affect the bio-oil yields within the tested ranges. The use of product gas as the fluidizing medium had a potential for increased bio-oil yields. PMID- 19560916 TI - Sulfonamides with the N-alkyl-N'-dialkylguanidine moiety as 5-HT7 receptor ligands. AB - A series of arylsulfonamides containing guanidine incorporated in the structure of secondary amines (piperidine, piperazine) was synthesized on SynPhase Lanterns and evaluated for 5-HT(1A), 5-HT(2A), and 5-HT(7) receptors. The results demonstrated that N-alkyl-N'-dialkylguanidines displayed good 5-HT(7)/5-HT(1A) selectivity and may be regarded as promising structural core for development of 5 HT(7) ligands. PMID- 19560917 TI - Tricyclic indole and dihydroindole derivatives as new inhibitors of soluble guanylate cyclase. AB - The synthesis of new tricyclic fused indole and dihydroindole derivatives and preliminary results from their in vitro inhibitory activity against soluble guanylate cyclase (sGC) are presented. PMID- 19560918 TI - Dopamine D3 receptor antagonists: the quest for a potentially selective PET ligand. Part one: lead identification. AB - The synthesis and SAR of a new series of potent and selective dopamine D(3) receptor antagonists is reported. PMID- 19560920 TI - Assessment of new 2'-O-acetalester protecting groups for regular RNA synthesis and original 2'-modified proRNA. AB - New base-labile acyloxymethyl groups were evaluated to protect 2'-OH functions of ribonucleotides for regular RNA synthesis in order to shorten the deprotection procedure upon ammonia. These same acetalester groups were assessed in 2' modified proRNA as biolabile 2'-protections removable by cell enzymes to generate parent RNA. Demasking of 2'-modified pro-uridylates was studied in cell extracts. PMID- 19560921 TI - Structure-Activity Relationship of novel phenylacetic CXCR1 inhibitors. AB - We reported recently the Structure-Activity Relationship (SAR) of a class of CXCL8 allosteric modulators. They invariably share a 2-arylpropionic moiety so far considered a key structural determinant of the biological activity. We show the results of recent SAR studies on a novel series of phenylacetic derivatives supported by a combined approach of mutagenesis experiments and conformational analysis. The results suggest novel insights on the fine role of the propionic/acetic chain in the modulation of CXCL8 receptors. PMID- 19560919 TI - Synthesis and evaluation of new endomorphin analogues modified at the Pro(2) residue. AB - Six new endomorphin analogues, incorporating constrained amino acids in place of native proline have been synthesized. Residues of (S)-azetidine-2-carboxylic acid (Aze), 3,4-dehydro-(S)-proline (Delta(3)Pro), azetidine-3-carboxylic acid (3Aze) and dehydro-alanine (DeltaAla) have been used to prepare [Delta(3)Pro(2)]EM-2 (1), [Aze(2)]EM-1 (2), [Aze(2)]EM-2 (3), [3Aze(2)]EM-1 (4), [3Aze(2)]EM-2 (5) and [DeltaAla(2)]EM-2 (6). Binding assays and functional bioactivities for mu- and delta-receptors are reported. The highest affinity, bioactivity and selectivity are shown by peptides 2 and 3 containing the Aze residue. PMID- 19560922 TI - Bis-anthracenyl isoxazolyl amides have enhanced anticancer activity. AB - Dimeric analogs of Anthracenyl Isoxazole Amides (AIMs) (the designation AIM is in honor of the memory of Professor Albert I. Meyers) were prepared and dimer 6 exhibited the highest efficacy to date for this class of anti-tumor compounds against the human glioma Central Nervous System cell line SNB-19. PMID- 19560923 TI - Conformationally restricted homotryptamines. Part 5: 3-(trans-2 aminomethylcyclopentyl)indoles as potent selective serotonin reuptake inhibitors. AB - A series of racemic 3-(trans-2-aminomethylcyclopentyl)indoles was synthesized and found to have potent binding to the human serotonin transporter (hSERT). The most active analog was synthesized stereospecifically and the active enantiomer was shown to have high affinity binding to hSERT. PMID- 19560924 TI - Functionalized 3-amino-imidazo[1,2-a]pyridines: a novel class of drug-like Mycobacterium tuberculosis glutamine synthetase inhibitors. AB - 3-Amino-imidazo[1,2-a]pyridines have been identified as a novel class of Mycobacterium tuberculosis glutamine synthetase inhibitors. Moreover, these compounds represent the first drug-like inhibitors of this enzyme. A series of compounds exploring structural diversity in the pyridine and phenyl rings have been synthesized and biologically evaluated. Compound 4n was found to be the most potent inhibitor (IC(50)=0.38+/-0.02 microM). This compound was significantly more potent than the known inhibitors, l-methionine-SR-sulfoximine and phosphinothricin. PMID- 19560925 TI - Visual deprivation leads to gait adaptations that are age- and context-specific: II. Kinematic parameters. AB - Significant differences exist between eyes open (EO) and eyes closed (EC) conditions in postural sway, self-selected dimensionless walking speed and duration of double support in both children and adults. The decrease in speed could be attributed to a decrease in dimensionless stride length. The question remains whether the slower gait is a reflection of fundamental changes in movement control in the absence of vision or it results from uncertainty or fear of falling. Studying the differences in foot and ankle kinematics between the two conditions may provide further information. In this study we compare the impact of visual deprivation on joint kinematics during locomotion in adults and children. Visual deprivation had a significant effect on the gait pattern of healthy children and adults. Differences included a more backward leaning position of the trunk, limited movement in the pelvis, reduced hip adduction during stance, increased flexion of the knee related to flat foot contact and reduced ankle plantar flexion at push-off. These differences indicate a more cautious walking strategy in the absence of vision, probably resulting from postural control problems. Although age also had a significant effect on gait in both EO and EC conditions, adults and children show little differences in their kinematic response to blindfolding. PMID- 19560926 TI - Intracranial infiltration by recurrent scalp dermatofibrosarcoma protuberans. AB - Dermatofibrosarcoma protuberans (DFSP) of the scalp is a rare soft tissue sarcoma. A 22-year-old male with DFSP, who had undergone several surgical excisions and radiotherapy, presented with a mild left lower-limb paresis. An MRI scan demonstrated a huge mass extending throughout the frontoparietal convexity. A macroscopically complete removal of the mass was performed. Despite this, there were several recurrences; thereafter, the patient underwent surgery, stereotactic radiosurgery and chemotherapy. The histological findings demonstrated a DFSP that both abutted and occasionally invaded the adjacent brain cortex, while extending along the Virchow-Robin spaces. Due to the highly infiltrating characteristics of DFSP, another approach, such as the use of molecular-targeted agents, will be required to significantly improve the clinical outcome. PMID- 19560927 TI - Urinary dysfunction in multiple sclerosis. AB - We aimed to determine the prevalence of urinary symptoms and complications in multiple sclerosis (MS) and to investigate the correlation of these symptoms and complications with urodynamic findings and disease characteristics (primary progressive, relapsing-remitting and secondary progressive). Fifty-two patients with MS were enrolled in the study. Patients were divided into three clinical types: primary progressive, relapsing-remitting and secondary progressive. Urological symptoms and complications were recorded. Each patient underwent a urodynamic investigation. Patients were evaluated for overall disability using the Expanded Disability Status Scale (EDSS). We found that 22 patients had primary progressive, 18 patients had relapsing-remitting and 12 patients had secondary progressive MS. Urgency was the most frequent urinary symptom (65%). Urinary infection was the most frequent urinary complication (15%). Detrusor hyperreflexia was found to be the main bladder dysfunction (27%). Detrusor sphincter dyssynergia was found in 25% of patients, and detrusor hyporeflexia was detected in 6%. No relationship was found between urinary symptoms and urodynamic abnormalities (p>0.05) and between urinary complications and urodynamic findings (p>0.05). No significant correlation was found between disease characteristics and urinary symptoms, urinary complications or urodynamic findings (p>0.05). We suggest that the assessment of urological symptoms and urodynamic evaluation is critical for evaluating quality of life in MS. PMID- 19560928 TI - Assessment of a clinical checklist in the diagnosis of fragile X syndrome in India. AB - Fragile X syndrome (FRAXA) is one of the most common forms of mental retardation. It is caused by the expansion of cytosine-guanine-guanine (CGG) repeats in the 5' untranslated region of the fragile X mental retardation 1 (FMR1) gene, located at Xq27.3. The number of CGG repeats in the FMR1 gene occurs in four distinct ranges: 2-50 (normal), 50-60 (gray zone), 60-200 (premutation), and > 200 (full mutation). When the number of CGG repeats exceeds 200, the gene becomes hypermethylated and transcriptionally silenced, which results in the loss of FMR protein and causes FRAXA. The key clinical features of FRAXA are mental retardation, macro-orchidism, long face, prominent jaw, connective tissue abnormalities, and behavioral problems. A modified 15-item checklist was used to assess the clinical features in 337 individuals (316 males and 21 females) who have mental retardation of unknown etiology. These patients were in institutions. Molecular diagnosis was performed using polymerase chain reaction and Southern blot analysis and revealed that 14 males were positive for FRAXA. Studies of the families of the affected males revealed an additional 11 affected males and 20 carrier females. Retrospective analysis of clinical features was performed in a total of 327 males and 41 females. Six clinical features were statistically significant in FRAXA individuals when compared to non-FRAXA individuals. These features were hyperactivity (p<0.05), poor eye contact (p<0.001), hyper extensibility of joints (p<0.001), large ears (p<0.001), macro-orchidism (p<0.001), and a family history of mental retardation (p<0.001). When a total score of 5 out of 15 was used as the threshold clinical score, 73.18% of the patients with total scores < 5 could be eliminated as FRAXA-negative patients, thereby improving the reliability of FRAXA testing using the clinical checklist. PMID- 19560929 TI - Postoperative intracerebral and intraventricular hemorrhages following removal of a chronic subdural hematoma. AB - We report a patient with multiple intracerebral and intraventricular hemorrhages after removal of bilateral chronic subdural hematomas (CSDH). Possible mechanisms include a sudden increase in cerebral blood flow within fragile cerebral vessels following decompression of CSDH, defective vascular autoregulation, and damage to the cerebral vascular tree. Clinical awareness of this complication and slow decompression of CSDH are stressed. PMID- 19560930 TI - An extended study of dimeric phenyl tropanes. AB - A series of dimeric phenyl tropanes consisting of two molecules of 4-chloro, 4 iodo or 4-(3-thiopheno)-phenyl tropane tethered together at the carboxylic acid moiety by a diamine or diol linker were prepared. The diamines used were a variety of linear, cyclic and aromatic diamines, while the diol tethered compounds were prepared by 'click' chemistry and contained a triazole in the linker. The new compounds were tested for binding to hDAT, hSERT and hNET. Amide linked chlorophenyl tropanes with an aromatic linker was found to be potent and selective DAT inhibitors with the best K(i) value for hDAT being 6nM. The ester linked halophenyl tropanes were more potent but displayed little selectivity in inhibition of monoamine transporter binding. Among the studied compounds an ester linker of 10 atoms between the tropane moieties gave the highest affinity. One monomeric phenyl tropane was made for comparison and was found to be less potent than the dimeric counterparts towards SERT and NET but remain highly active against DAT. Dimeric thiophenophenyl tropanes were in general found to be comparatively poor monoamine transporter binders, but significant gains of affinity of up to 45-fold could be achieved with selected dimeric chlorophenyl tropanes compared to the parent monomer. This observation implies that a secondary binding site that has affinity for phenyl tropanes, most likely the putative S2 site, is located within 13A of the primary central S1 binding site. PMID- 19560931 TI - Synthesis and biological evaluation of new 4-carboxyl quinoline derivatives as cyclooxygenase-2 inhibitors. AB - A group of 4-carboxyl quinoline derivatives possessing a methylsulfonyl COX-2 pharmacophore at the para position of the C-2 phenyl ring were designed and synthesized as selective COX-2 inhibitors. In vitro COX-1/COX-2 structure activity relationships were determined by varying the substituents on the C-7 and C-8 quinoline ring. Among the 4-carboxyl quinolines, 7,8,9,10-tetrahydro-2-(4 (methyl sulfonyl)phenyl)benzo[h]quinoline-4-carboxylic acid (9e) was identified as potent and high selective COX-2 inhibitor (COX-2 IC(50)=0.043microM; selectivity index>513) that was more potent than the reference drug celecoxib (COX-2 IC(50)=0.060microM; SI=405). A molecular modeling study where 9e was docked in the binding site of COX-2 showed that the p-MeSO(2) substituent on the C-2 phenyl ring is oriented in the vicinity of the COX-2 secondary pocket (Arg513, Phe518 and Val523) and the carboxyl group can interact with Arg120. The structure activity data acquired indicate that the presence of lipophilic substituents on the C-7 and C-8 quinoline ring is important for COX-2 inhibitory activity. PMID- 19560932 TI - Activation analysis: a method for the future. PMID- 19560933 TI - Unexpected stent thrombus after minimally invasive direct coronary artery bypass in hybrid re-vascularisation. AB - The progress in anti-platelet therapy and percutaneous coronary intervention led to reconfigure indications of hybrid re-vascularisation. However, there are still some controversies over indication, timing and patient management during the procedure. The case discussed here is a patient who was diagnosed with myocardial infarction and treated with hybrid re-vascularisation. The patient underwent stent insertion followed by bypass surgery. After the hybrid procedure, the patient was stable but eventually died on the 12th day after the surgery owing to unexpected stent thrombosis. We discuss the current controversy over hybrid re vascularisation, variables that can affect the outcome and the requirement for establishing accurate logistics based on our case. PMID- 19560934 TI - Hyperchloraemic metabolic acidosis slows recovery in children with diabetic ketoacidosis: a retrospective audit. AB - INTRODUCTION: Hyperchloraemic metabolic acidosis (HMA) can occur in diabetic ketoacidosis (DKA), from urinary loss of bicarbonate precursors as ketones, or iatrogenically from chloride administration. OBJECTIVE: To determine whether children with DKA given normal saline developed HMA, and whether HMA delayed their recovery. SETTING: 13 Bed combined Paediatric Intensive Care/High Dependency Unit. METHODS: Retrospective analysis of the venous biochemistry of 59 admissions with DKA, recording the times to recovery from acidosis and normalisation of anion gap, and total intravenous chloride load. RESULTS: Twenty nine (49%) were newly diagnosed diabetics. The median age was 12 (interquartile range, IQR 8.2-15.4) years. The initial pH in 23 (39%) was <7.1. The median times to achieve pH>7.3, bicarbonate>15mmol/l and anion gap<16.1 were 14.2h (IQR 8.6 20.1), 12.9h (IQR 8.6-20.0) and 10.7h (IQR 8.2-15.0) respectively. For individual patients, the median difference between recovery times for bicarbonate and anion gap was 0.18h (IQR 0-5.3), p=0.0005. However, in 14 patients (24%), the difference was >6h. These patients did not differ significantly in age or initial pH but had a lower initial bicarbonate (median 5 versus 7.8mmol/l, p=0.002), narrower anion gap (median 29.5 versus 31.6mmol/l, p=0.038), and took longer to normalise the bicarbonate: median 26.1 versus 10.5h, p<0.0001. They tended to be newly diagnosed presentations. CONCLUSION: The anion gap (AG) normalises earlier than bicarbonate in children with DKA treated with normal saline, and children with persisting HMA recover from acidosis more slowly. PMID- 19560935 TI - Grape-seed procyanidins modulate inflammation on human differentiated adipocytes in vitro. AB - Flavonoids are functional constituents of many fruits and vegetables. Procyanidins are flavonoids with an oligomeric structure, and it has been shown that they can improve the pathological oxidative state of a diabetic situation. To evaluate whether procyanidins can modulate inflammation, an event strongly associated with obesity, diabetes and insulin resistance states, we used human adipocytes (SGBS) and macrophage-like (THP-1) cell lines and administered an extract of grape-seed procyanidins (GSPE). THP-1 and SGBS cells pre-treated with GSPE showed a reduction of IL-6 and MCP-1 expression after an inflammatory stimulus. GSPE stimuli alone modulate adipokine (APM1 and LEP) and cytokine (IL-6 and MCP-1) gene expression. GSPE partially inhibited NF-kappaB translocation to the nucleus in both cell lines. These preliminary findings demonstrate that GSPE reduces the expression of IL-6 and MCP-1 and enhances the production of the anti inflammatory adipokine adiponectin suggesting that may have a beneficial effect on low-grade inflammatory diseases such obesity and type 2 diabetes. PMID- 19560937 TI - Importance of angiographic visualization of round ligament arteries in women evaluated for intractable vaginal bleeding after uterine artery embolization. AB - PURPOSE: To determine the incidence of angiographic visualization and the clinical significance of round ligament arteries in patients who present with intractable vaginal bleeding. MATERIALS AND METHODS: A review of 113 patients (age range, 20-67 years) who underwent pelvic angiography for intractable vaginal bleeding between June 1992 and May 2008 was retrospectively performed. It was recorded whether round ligament artery was visualized on pelvic aortography after uterine artery embolization (UAE). The medical records of the patients were reviewed to analyze the final clinical outcome. The Fisher exact test was used to correlate persistent vaginal bleeding after UAE with visualization of round ligament arteries. RESULTS: Of 111 patients who underwent UAE, 42 patients (postpartum bleeding, n = 40; postabortion bleeding, n = 2) had at least one visible round ligament artery on postembolization pelvic aortography. Ten patients received round ligament artery embolization. Persistent vaginal bleeding after adequate UAE was observed more commonly in patients whose round ligament artery was seen on postembolization pelvic aortography (P = .007). CONCLUSIONS: Round ligament arteries are commonly visualized in patients who present with postpartum bleeding and should be investigated when there is persistent bleeding, even after adequate UAE. PMID- 19560936 TI - Fragmentation mechanisms of oxidized peptides elucidated by SID, RRKM modeling, and molecular dynamics. AB - The gas-phase fragmentation reactions of singly charged angiotensin II (AngII, DR(+)VYIHPF) and the ozonolysis products AngII+O (DR(+)VY*IHPF), AngII+3O (DR(+)VYIH*PF), and AngII+4O (DR(+)VY*IH*PF) were studied using SID FT-ICR mass spectrometry, RRKM modeling, and molecular dynamics. Oxidation of Tyr (AngII+O) leads to a low-energy charge-remote selective fragmentation channel resulting in the b(4)+O fragment ion. Modification of His (AngII+3O and AngII+4O) leads to a series of new selective dissociation channels. For AngII+3O and AngII+4O, the formation of [MH+3O](+)-45 and [MH+3O](+)-71 are driven by charge-remote processes while it is suggested that b(5) and [MH+3O](+)-88 fragments are a result of charge-directed reactions. Energy-resolved SID experiments and RRKM modeling provide threshold energies and activation entropies for the lowest energy fragmentation channel for each of the parent ions. Fragmentation of the ozonolysis products was found to be controlled by entropic effects. Mechanisms are proposed for each of the new dissociation pathways based on the energies and entropies of activation and parent ion conformations sampled using molecular dynamics. PMID- 19560938 TI - G2 inferior vena cava filter: retrievability and safety. AB - PURPOSE: To assess the retrievability of the G2 inferior vena cava (IVC) filter and factors influencing the safety and technical success of retrieval. MATERIALS AND METHODS: From October 2006 through June 2008, G2 IVC filters were placed in 140 consecutive patients who needed prophylaxis against pulmonary embolism (PE). General indications for filter placement included history of thromboembolic disease (n = 98) and high risk for PE (n = 42); specific indications included contraindication to anticoagulation (n = 120), prophylaxis in addition to anticoagulation (n = 16), and failure of anticoagulation (n = 4). Filter dwell time, technical success of filter retrieval, and complications related to placement or retrieval were retrospectively evaluated in patients who underwent filter removal. RESULTS: Twenty-seven attempts at G2 filter removal were made in 26 patients (12 men; age range, 24-88 years; mean age, 55.4 y) after a mean period of 122 days (range, 11-260 d). Data were collected retrospectively with institutional review board approval. Filter removal was successful in all 27 attempts (100%). Tilting of the filter (> or =15 degrees ) occurred in five cases (18.5%), with probable filter incorporation into the right lateral wall of the IVC in one. Other complications of retrieval such as filter thrombosis, significant filter migration, filter fracture, and caval occlusion were not observed. CONCLUSIONS: G2 IVC filter retrieval has a high technical success rate and a low complication rate. Technical success appears to be unaffected by the dwell time within the reported range. PMID- 19560939 TI - Penetration of Gunther Tulip filter struts through an introducer sheath: case report and safety concerns. AB - The authors present a patient who had a Gunther-Tulip inferior vena cava filter placed under fluoroscopic guidance. The filter struts were seen to penetrate through the introducer sheath at deployment. This was believed to be secondary to a kink in the sheath at the site of venous entry and was due to the patient's nuchal obesity as well as his inability to turn his head to the opposite side. The introducer sheath was then placed through a long reinforced metal sheath through which the filter was then placed without complication. PMID- 19560940 TI - Efficacy of intrapleural tissue-type plasminogen activator in the treatment of loculated parapneumonic effusions. AB - PURPOSE: To assess the feasibility and effectiveness of intrapleural recombinant tissue-type plasminogen activator (r-tPA) in the treatment of loculated parapneumonic effusions (PPEs). MATERIALS AND METHODS: A single-arm prospective study of 25 consecutive patients with loculated PPEs was analyzed. All patients received 6-mg doses of intrapleural r-tPA on a defined schedule via a thoracostomy tube. The volume of output from the tubes was recorded and analysis of the fluid composition performed. Follow-up was both clinical and radiographic, with all patients undergoing pre- and postprocedural computed tomography. RESULTS: Eighteen of the 25 patients (72%) required no additional intervention and had a complete clinical and radiographic response with the fibrinolytic therapy. Seven patients (28%) were treated with video-assisted thoracoscopic surgery, but no patient required thoracotomy for total decortication. There were no hemorrhagic complications. CONCLUSIONS: Intrapleural r-tPA is effective in the treatment of loculated PPEs. It can be performed safely and in some patients may avoid the need for additional surgical intervention. PMID- 19560942 TI - Renal artery aneurysm and arteriovenous fistula associated with fibromuscular dysplasia: successful treatment with detachable coils. AB - Renal artery aneurysms are more common in patients with fibromuscular dysplasia (FMD). Although these aneurysms have traditionally been managed surgically, endovascular techniques are playing a larger role. Many treatment methods involve the use of stent-grafts to exclude the aneurysm or bare stents to protect the main renal artery while coil embolization is performed through the interstices. Herein, the authors present a patient with an arteriovenous fistula secondary to FMD that was managed entirely with use of detachable coils, which allowed preservation of the uninvolved renal parenchyma. PMID- 19560941 TI - Development of a VX2 pancreatic cancer model in rabbits: a pilot study. AB - PURPOSE: An animal model of pancreatic cancer that is large enough to permit imaging and catheterization would be desirable for interventional radiologists to develop novel therapies for pancreatic cancer. The purpose of this study was to test the hypothesis that the VX2 rabbit model of pancreatic cancer could be developed as a suitable platform to test future interventional therapies. MATERIALS AND METHODS: The authors implanted and grew three pancreatic VX2 tumors per rabbit in six rabbits. Magnetic resonance (MR) imaging was performed at 2 weeks to confirm tumor growth. At 3 weeks, the authors selectively catheterized the gastroduodenal artery under guidance of x-ray digital subtraction angiography (DSA). T2-weighted anatomic imaging, diffusion-weighted MR imaging, and transcatheter intraarterial perfusion (TRIP) MR imaging were then performed. After imaging, tumors were confirmed at necropsy and histopathologically. Tumor sizes at 2 and 3 weeks were compared with a paired t test (P = .05). RESULTS: VX2 pancreatic tumors were grown in all six rabbits. The difference between tumor sizes at 2 and 3 weeks (1.29 cm +/- 0.39 vs 1.91 cm +/- 0.50, respectively) was significant (P < .001). All tumors were confirmed to be located within pancreatic tissue via histopathologic analysis. DSA and TRIP MR imaging were successful in five rabbits. Diffusion-weighted and anatomic MR imaging were successful in all six rabbits. CONCLUSIONS: The VX2 rabbit model of pancreatic cancer is feasible, as verified by imaging and pathologic correlation, and may be a suitable platform to test future interventional therapies. PMID- 19560944 TI - Suture anchor loading after rotator cuff repair: effects of an additional lateral row. AB - HYPOTHESIS: Our initial hypothesis was that the medial row of double-row rotator cuff repair techniques would bear most of the load on the repaired cuff. MATERIALS AND METHODS: Six cadaver shoulders underwent simulated rotator cuff repairs using sequential single row, double-row, and suture-bridge repair techniques. Suture tensions at each anchor were measured for several static, simulated shoulder positions by specially designed, instrumented anchors. RESULTS: Significantly greater suture tensions were measured in the anchors in a single row repair construct than either the double row repair or suture bridge repair construct (P < .001). In the double-row and suture bridge techniques, there was no apparent difference in the loads born by the medial and lateral row anchors. Shoulder abduction from 45 degrees to 60 degrees had little effect on anchor tensions; 45 degrees internal and external rotation significantly (P = .032) increased loads on the anterior and posterior anchors by at least 125%. DISCUSSION: Forces are transmitted through the entire portion of the tendon at its humeral fixation, loading the lateral anchors as well as the medial row for the techniques studied. This "load sharing" can explain the higher fixation strengths of double row techniques seen experimentally. CONCLUSION: The magnitude and distribution of anchor suture tensions could have important implications for lateral row fixation devices and post-operative positioning and activity. LEVEL OF EVIDENCE: Basic Science. PMID- 19560945 TI - Endothelial progenitor cells as a therapeutic option in peripheral arterial disease. AB - BACKGROUND: Endothelial progenitor cells (EPC) are a subpopulation of bone-marrow mononuclear cells that are capable of generating new blood vessels in areas of ischaemia or infarction. This review examines the regenerative potential of EPC to ameliorate peripheral ischaemia. METHODS: An online search was done using OVID Medline Search, PubMed, and Cochrane Review Database, for all reviews and original articles in English concerning progenitor or bone-marrow mononuclear cells. RESULTS AND CONCLUSION: There are many controversies in EPC research, especially in the areas of identification, characterization, and therapeutic use. Both animal and human studies have shown benefits from using EPC to combat peripheral arterial and cerebrovascular disease. To bring EPC into wider clinical use, larger controlled clinical trials and better methods of augmenting EPC function and lifespan are required. Until then EPC should be used under robust trial conditions with ethical approval. PMID- 19560946 TI - Plasma fibrinogen and D-dimer concentrations are associated with the presence of abdominal aortic aneurysm: a systematic review and meta-analysis. AB - OBJECTIVES: To summarise the present evidence for an association between circulating fibrinogen or D-dimer and presence of abdominal aortic aneurysm (AAA) presence. DESIGN: MEDLINE database was searched to identify all case-control studies that compared plasma fibrinogen or D-dimer concentrations between patients with AAA and subjects without AAA. For each study, data regarding fibrinogen or D-dimer concentrations in both the AAA and control groups were used to generate mean differences (MDs) and 95% confidence intervals (CIs). Study specific estimates were combined using inverse variance-weighted average of logarithmic MDs in both fixed- and random-effects models. RESULTS: Our search identified 10 eligible studies including 834 cases with AAA and 6971 controls without AAA for fibrinogen and six studies including 264 patients with AAA and 403 subjects without AAA for D-dimer. Pooled analysis demonstrated significantly higher fibrinogen (fixed-effects MD, 0.37gl(-1); 95% CI: 0.30-0.44gl(-1)) and D dimer (random-effects MD: 415.36ngml(-1); 95% CI: 128.97-701.76ngml(-1)) concentrations in the AAA group than those in the control group. CONCLUSIONS: We found that plasma fibrinogen and D-dimer concentrations are likely to be higher in cases with AAA than control subjects. Higher plasma fibrinogen and D-dimer concentrations may be associated with the presence of AAA. PMID- 19560947 TI - Visceral venous aneurysms: clinical presentation, natural history and their management: a systematic review. AB - AIM: Aneurysms of the visceral veins are considered rare clinical entities. The aim is to assess their clinical presentation, natural history and management. METHODS: An electronic search of the pertinent English and French literature was undertaken. All studies reporting on aneurysms of visceral veins were considered. Cases describing patients with arterial-venous fistulae and extrahepatic or intra hepatic portosystemic venous shunts were excluded. RESULTS: Ninety-three reports were identified, including 176 patients with 198 visceral venous aneurysms. Patients' age ranges from 0 to 87 years, and there is no apparent male/female preponderance. The commonest location of visceral venous aneurysms is the portal venous system (87 of 93 reports, 170 of 176 patients, 191 of 198 aneurysms). Aneurysms of the renal veins and inferior mesenteric vein are also described. Portal system venous aneurysms were present with abdominal pain in 44.7% of the patients, gastrointestinal bleeding in 7.3%, and are asymptomatic in 38.2%. Portal hypertension is reported in 30.8% and liver cirrhosis in 28.3%. Thrombosis occurred in 13.6% and rupture in 2.2% of the patients. Adjacent organ compression is reported in 2.2% (organs compressed: common bile duct, duodenum, inferior vena cava). The management ranged from watchful waiting to intervention. In 94% of the cases, aneurysm diameter remained stable and no complications occurred during follow-up. In most of the cases, indications for operation were symptoms and complications. Six cases of renal vein aneurysm are reported; three of them were asymptomatic. Three of these patients were treated surgically. CONCLUSION: The most frequent location of visceral venous aneurysms is the portal venous system. They are often associated with cirrhosis and portal hypertension. They may be asymptomatic or present with abdominal pain and other symptoms. Watchful waiting is an appropriate treatment, except when complications occur. Most common complications are aneurysm thrombosis and rupture. Other visceral venous aneurysms are extremely rare. PMID- 19560948 TI - The prevalence of polyvascular disease in patients referred for peripheral arterial disease. AB - OBJECTIVE: To objectively assess the presence of polyvascular disease in patients with peripheral arterial disease and its relation to inflammation and clinical risk factors. METHODS: A total of 431 vascular surgery patients (mean age 68 years, men 77%) with atherosclerotic disease were enrolled. The presence of atherosclerosis was assessed using ultrasonography. Affected territories were defined as: (1) carotid, stenosis of common or internal carotid artery of >or=50%, (2) cardiac, left ventricular wall motion abnormalities, (3) abdominal aorta, diameter >or=30 mm and (4) lower limb, ankle-brachial pressure index <0.9. Cardiovascular risk factors and high-sensitivity C-reactive protein (hs-CRP) levels were noted in all. RESULTS: One vascular territory was affected in 29% of the patients, whereas polyvascular disease was found in 71%: two affected territories in 45%, three in 23% and four in 3% of patients. Levels of hs-CRP increased with the number of affected vascular territories (p<0.001). Multivariable logistic regression analysis showed age >or=70 years, male gender, body mass index (BMI)>or=25 kg m(-2), and hs-CRP to be independently associated with polyvascular disease. CONCLUSION: Polyvascular disease is a common condition in patients who have undergone vascular surgery. The level of systemic inflammation, reflected by hs-CRP levels, is moderately associated with the extent of polyvascular disease. PMID- 19560949 TI - Comment on "Early carotid endarterectomy after intravenous thrombolysis for acute ischaemic stroke, Bartolia MA et al. May 2009". PMID- 19560950 TI - Regulation of matrix contraction in chronic venous disease. AB - OBJECTIVE: The role of TGF-beta(1) in venous ulcer healing and the signalling cascades regulating dermal fibroblast function are poorly understood. To elucidate these processes, we hypothesized that TGF-beta(1) facilitates wound healing by increasing chronic venous insufficiency (CVI) induced matrix contraction via intracellular cross-talk between TGF-beta(1) and the ERK-1/2 MAP kinase signalling cascades. METHODS: Fibroblasts isolated from calf biopsies (LC) of patients with different severity of CVI (CEAP, Clinical Etiological Anatomical Pathological classes) were seeded into 200 microl collagen gels under isometric conditions. Fibroblasts from neonatal foreskins (HS68), non-CVI patients (NC), and the ipsilateral normal thigh of each CVI patient (LT) served as controls. Thirteen patients with CVI (class 2, n=5; class 4, n=5; class 6, n=3) and 2 non CVI controls (NC, n=2) were included in the study. All experimental conditions were determined by dose-response and time-course experiments. Gels were cultured with/without 0.1 ng/ml TGF-beta(1) and with/without 50 microM PD98059 (MEK and downstream-MAPK inhibitor). Additional patient fibroblasts were transfected with constitutively active Ras (pCMV-Ras) or an empty vector (pCMV-beta) with/without 0.1 ng/ml TGF-beta(1) and with/without 50 microm PD98059. The collagen gels were released after 4 days and the percent contraction was determined by area measurements using image analysis. Differences in alpha-smooth muscle actin (alpha-SMA) and ERK-1/2 MAPK (phosphorylated and total) protein levels were analyzed with western blotting. RESULTS: Gels seeded with CVI fibroblasts contracted more than HS68, NC and LT fibroblasts. Inhibition of MAPK and/or stimulation with TGF-beta(1) increased the contraction of LC gels compared to unstimulated controls. Agonist induced gel contraction correlated with CVI disease severity. alpha-SMA protein expression in LC fibroblasts increased with MAPK inhibition with/without TGF-beta(1) stimulation, and correlated with the degree of gel contraction. Transfection with pCMV-Ras (activator of ERK-1/2) inhibited gel contraction; this inhibition was not reversed by addition of TGF beta(1). Transfection with the pCMV-beta empty vector had no effect on gel contraction. CONCLUSIONS: TGF-beta1 stimulation of CVI patient fibroblasts grown in 3D collagen gels results in conversion to a contractile phenotype through upregulation of alpha-SMA, and in enhanced gel contraction. Inhibition of MAPK further increases gel contraction, while Ras activation of ERK-1/2 inhibits TGF beta1-induced gel contraction. These responses correlate with increasing CEAP severity. CVI fibroblast mediated gel contraction is therefore regulated through cross-talk between the ERK-1/2 MAPK and TGF-beta(1) signalling cascades. These data identify potentially clinically relevant therapeutic molecular targets that could enhance matrix contraction and thereby improve venous ulcer wound healing. PMID- 19560951 TI - Homocysteine and peripheral arterial disease: systematic review and meta analysis. AB - OBJECTIVE: To evaluate homocysteine (Hcy) levels in patients with peripheral arterial disease (PAD) as compared to unaffected controls, and to review the clinical effects of therapy aimed at lowering homocysteine in PAD patients. METHODS: MEDLINE, EMBASE and Cochrane databases were searched from 1950 to December 2007. We selected observational studies and trials that evaluated Hcy levels in patients with PAD compared to unaffected controls. We also included trials on the effect of Hcy-lowering therapy (folate supplementation) in PAD patients. Continuous outcomes were pooled in a random effects meta-analysis of the weighted mean difference between comparator groups. RESULTS: We retrieved 33 potentially suitable articles from our search. Meta-analysis of 14 relevant studies showed that Hcy was significantly elevated (pooled mean difference +4.31micromoll; 95% C.I. 1.71, 6.31, p<0.0001 with significant heterogeneity) in patients with PAD compared to controls. As all 14 studies consistently demonstrated raised plasma Hcy levels in PAD patients, the significant heterogeneity in this meta-analysis probably arises from differences in the degree of Hcy elevation. The effect of folate supplementation on PAD was tested in eight clinical trials but clinically important end points were inconsistently reported. CONCLUSION: Patients with PAD have significantly higher Hcy levels than unaffected controls. However, we did not find any robust evidence on clinically beneficial effects of folate supplementation in PAD. PMID- 19560952 TI - Haemostatic abnormalities and clinical findings in Vipera palaestinae-envenomed dogs. AB - The venomous viper Vipera palaestinae (Vp) is responsible for most envenomations in humans and animals in Israel. Its venom contains proteases, haemorrhagins, L amino acid oxidase and phospholipase A2 but its effects on haemostasis have yet to be characterised. This prospective study aimed to characterise haemostatic abnormalities in Vp-envenomed dogs from presentation to discharge or death, and their association with mortality. Samples from 39 Vp-envenomed dogs were collected periodically and examined for haematology, prothrombin time (PT), activated partial thromboplastin time (aPTT), antithrombin activity (ATA), fibrinogen level and D-dimer concentration. All dogs presented with severe local signs and most (79%) had systemic signs. Six dogs (15%) died. Haemostatic abnormalities were present in 37/39 dogs. Increased D-dimer concentrations were detected in 28/31 dogs. Disseminated intravascular coagulation was diagnosed in 10 dogs and in all non-survivors. Platelet and leucocyte counts at presentation, maximum PT and aPTT, and minimum ATA during hospitalisation were significantly different between survivors and non-survivors and were good predictors of the outcome. The results show that hypercoagulability, consumption and derangement of haemostasis are common in Vp-envenomed dogs and are associated with mortality. Haemostasis should be closely monitored in such dogs. PMID- 19560953 TI - Immunoproteomic analysis of whole proteins from male and female adult Haemonchus contortus. AB - Whole proteins of male and female adult Haemonchus contortus were analysed by immunoproteomic techniques. Approximately 662 and 680 spots were detected on proteome maps of male and female nematodes, respectively, stained with Coomassie brilliant blue G-250. There were 609 shared spots. Approximately 193 and 196 spots were recognised on Western blot maps of male and female nematodes, respectively, using antiserum from naturally infected goats as the source of primary antibodies. There were 129 gender-specific spots in male nematodes and 132 in females. Twenty-three shared immunogenic spots were identified by MALDI TOF or MALDI-TOF-TOF mass spectrometry. These proteins included glutamate dehydrogenase (GDH), homologues of Dim-1, actin, globin-like excretory/secretory protein F6, glutathione S-transferase (GST), ATPase and glyceraldehyde-3 phosphate dehydrogenase. GDH and GST have been identified as immunogenic proteins of H. contortus previously, whereas the other proteins are newly recognised immunogenic proteins in this nematode. PMID- 19560955 TI - Clear cell sarcoma: a rare entity within melanoma. PMID- 19560954 TI - Hyperhomocysteinemia is associated with hypertriglyceridemia in mice with methylenetetrahydrofolate reductase deficiency. AB - Hyperhomocysteinemia (HHcy) can result from genetic or nutritional disturbances in folate metabolism. The most common genetic cause of mild HHcy is methylenetetrahydrofolate reductase (MTHFR) deficiency. To explore interactions between HHcy and lipid metabolism in atherogenesis, we measured plasma homocysteine (Hcy), triglycerides and cholesterol in Mthfr(+/+) and Mthfr(+/-) mice on C57BL/6 and BALB/c backgrounds, fed control or folate-deficient diets. We also crossed ApoE(C57)(-/-) mice with Mthfr(C57)(+/-) and Mthfr(BALB/c)(+/-) mice, and examined the same plasma variables as well as lipid accumulation in aortic sinus and whole aorta. Mthfr(+/-) mice had significantly higher plasma Hcy and plasma triglycerides relative to Mthfr(+/+) mice. A significant positive correlation was observed between plasma Hcy and plasma triglycerides in all mice. Mthfr(+/-) mice had lower plasma ApoA-IV protein levels which could reduce clearance of triglyceride-rich lipoproteins from the circulation. In the double mutant experiments, plasma Hcy was higher in Mthfr(+/-) compared with Mthfr(+/+) in ApoE(C57)(-/-)/Mthfr(C57) and ApoE(C57)(-/-)/Mthfr(BALB/c) mice. Triglycerides in female ApoE(C57)(-/-)/Mthfr(BALB/c)(+/-) mice were higher than those in ApoE(C57)(-/-)/Mthfr(BALB/c)(+/-) mice and correlated positively with Hcy. ApoE(C57)(-/-)/Mthfr(C57)(+/-) male mice had more lipid deposition in aortic sinus and whole aorta compared with ApoE(C57)(-/-)/Mthfr(c57)(+/+) mice. Our results suggest that HHcy is associated with hypertriglyceridemia and that MTHFR deficiency may exacerbate lipid accumulation in ApoE deficiency. PMID- 19560956 TI - Methotrexate: still the anchor drug in RA treatment. PMID- 19560957 TI - Degradation behavior and products of malathion and chlorpyrifos spiked in apple juice by ultrasonic treatment. AB - Apple juice (13 degrees Brix) spiked with malathion and chlorpyrifos (2-3 mg l( 1) of each compound) was treated under different ultrasonic irradiations. Results showed that ultrasonic treatment was effective for the degradation of malathion and chlorpyrifos in apple juice, and the output power and treatment time significantly influenced the degradation of both pesticides (p<0.05). The maximum degradations were achieved for malathion (41.7%) and chlorpyrifos (82.0%) after the ultrasonic treatment at 500 W for 120 min. The degradation kinetics of both pesticides were fitted to the first-order kinetics model well (R(2)>or=0.90). The kinetics parameters indicated that chlorpyrifos was much more labile to ultrasonic treatment than malathion. Furthermore, malaoxon and chlorpyrifos oxon were identified as the degradation products of malathion and chlorpyrifos by gas chromatography-mass spectrometry (GC-MS), respectively. The oxidation pathway through the hydroxyl radical attack on the P=S bond of pesticide molecules was proposed. PMID- 19560958 TI - Regional frequency variation during human ventricular fibrillation. AB - Quantifying the regional frequency variation in ventricular fibrillation (VF) may lead to focal strategies in treating human VF. We hypothesized that during human VF there are quantifiable regional frequency variations in the ventricles and they relate to underlying fixed myocardial substrate. In eight myopathic human hearts, we studied 35 VF episodes. The electrograms during VF were acquired simultaneously from the epicardium and endocardium using 2 electrode arrays each consisting of 112 electrodes. Regional characterization was performed using a ratio parameter derived from the dominant frequency analysis of the electrograms. The findings were related to the anatomical substrate using bipolar voltage maps. The results of the analysis indicate that LV had a larger dominant frequency (DF) span than RV (p=0.0111) while there was no significant difference (p=0.1488) in the DF span between LV freewall (FW) and septum (SE). Correlation of areas of abnormal myocardium with the dominant frequency feature matched only in 50% of the cases indicating that ion channel heterogeneity and time-varying physiological factors may play an important role in maintaining VF. PMID- 19560959 TI - Rivers and mountains may change, human nature does not! (traditional Chinese saying). PMID- 19560961 TI - Revised vibrational band assignments for the experimental IR and Raman spectra of 2,3,4-trifluorobenzonitrile based on ab initio, DFT and normal coordinate calculations. AB - In the present study, a systematic vibrational spectroscopic investigation for the experimental IR and Raman spectra of 2,3,4-trifluorobenzonitrile (TFB), aided by electronic structure calculations has been carried out. The electronic structure calculations -ab initio (RHF) and hybrid density functional methods (B3LYP) - have been performed with 6-31G* basis set. Molecular equilibrium geometries, electronic energies, IR intensities, harmonic vibrational frequencies, depolarization ratios and Raman activities have been computed. The results of the calculations have been used to simulate IR and Raman spectra for TFB that showed excellent agreement with the observed spectra. Potential energy distribution (PED) and normal mode analysis have also been performed. The assignments proposed based on the experimental IR and Raman spectra have been reviewed. A complete assignment of the observed spectra has been proposed. PMID- 19560960 TI - Coordination of Ce(III) and Nd(III) with pentaethylene glycol in the presence of picrate anion: spectroscopic and X-ray structural studies. AB - (1)H NMR evidence for direct coordination between the Ln(III) ion and the oxygen atoms of the pentaethylene glycol (EO5) ligand and the picrate anion (Pic) in [Ln(Pic)(2)(EO5)][Pic] {Ln=Ce and Nd} complexes are confirmed by single X-ray diffraction. No dissociation of Ln-O bonds in dimethyl sulfoxide-d solution was observed in NMR studies conducted at different temperatures ranging 25-100 degrees C. The Ln(III) ion was chelated to nine oxygen atoms from the EO5 ligand in a hexadentate manner and the two Pic anions in each bidentate and monodentate modes. Both compounds are isostructural and crystallized in monoclinic with space group P2(1)/c. Coordination environment around the Ce1 and Nd1 atoms can be described as tricapped trigonal prismatic and monocapped square antiprismatic geometries, respectively. The crystal packing of the complexes have stabilized by one dimensional (1D) chains along the [001] direction to form intermolecular O Hcdots, three dots, centeredO and C-Hcdots, three dots, centeredO hydrogen bonding. The molar conductance of the complexes in DMSO solution indicated that both compounds are ionic. The complexes had a good thermal stability. Under the UV-excitation, these complexes exhibited the red-shift emission. PMID- 19560962 TI - Enhanced identification of eligibility for depression research using an electronic medical record search engine. AB - PURPOSE: Electronic medical records (EMRs) have become part of daily practice for many physicians. Attempts have been made to apply electronic search engine technology to speed EMR review. This was a prospective, observational study to compare the speed and clinical accuracy of a medical record search engine vs. manual review of the EMR. METHODS: Three raters reviewed 49 cases in the EMR to screen for eligibility in a depression study using the electronic medical record search engine (EMERSE). One week later raters received a scrambled set of the same patients including 9 distractor cases, and used manual EMR review to determine eligibility. For both methods, accuracy was assessed for the original 49 cases by comparison with a gold standard rater. RESULTS: Use of EMERSE resulted in considerable time savings; chart reviews using EMERSE were significantly faster than traditional manual review (p=0.03). The percent agreement of raters with the gold standard (e.g. concurrent validity) using either EMERSE or manual review was not significantly different. CONCLUSIONS: Using a search engine optimized for finding clinical information in the free-text sections of the EMR can provide significant time savings while preserving clinical accuracy. The major power of this search engine is not from a more advanced and sophisticated search algorithm, but rather from a user interface designed explicitly to help users search the entire medical record in a way that protects health information. PMID- 19560963 TI - An algorithm for the typing of enteroviruses and correlation to serotyping by viral neutralization. AB - BACKGROUND: Human enteroviruses (HEVs) are common pathogens which cause a broad spectrum of illnesses ranging from asymptomatic infection to acute myocarditis and aseptic meningitis. The neutralization assay for serotype determination is labor-intensive and time-consuming. There is a need for a methodology that is more rapid and widely accessible. OBJECTIVES: Our goals were to develop an algorithm to type enteroviruses which combines both serologic typing, based on indirect immunofluorescence assay (IFA) using type-specific monoclonal antibodies (mAbs) and genotyping, by DNA sequence analysis and to assess the correlation of both IFA and genotyping to traditional viral neutralization by type-specific antisera. STUDY DESIGN: Clinical specimens initially determined to be enterovirus positive by nucleic acid detection were grown in cell culture and typed using mAbs. Specimens that could not be typed by mAbs were subject to molecular analysis. Genotyping was performed by a combination of either a primary or semi nested RT-PCR for a region within VP3/VP1 and followed by direct DNA sequencing of PCR products. Database homology comparisons and phylogenetic analysis were performed based on a defined region (303 nt) within the VP1 gene. RESULTS: We inoculated 134 enterovirus nucleic acid amplification-positive specimens into culture and 115 (86%) of these isolates were successfully typed by this algorithm. We have demonstrated a strong correlation between serotyping by viral neutralization to both IFA by type-specific mAbs and genotyping. CONCLUSIONS: Typing of human enteroviruses can be effectively performed using an integration of antibody-based and molecular methods. PMID- 19560964 TI - Human KI and WU polyomavirus infection in immunocompromised subjects. PMID- 19560965 TI - Visual stimuli for the P300 brain-computer interface: a comparison of white/gray and green/blue flicker matrices. AB - OBJECTIVE: The white/gray flicker matrix has been used as a visual stimulus for the so-called P300 brain-computer interface (BCI), but the white/gray flash stimuli might induce discomfort. In this study, we investigated the effectiveness of green/blue flicker matrices as visual stimuli. METHODS: Ten able-bodied, non trained subjects performed Alphabet Spelling (Japanese Alphabet: Hiragana) using an 8 x 10 matrix with three types of intensification/rest flicker combinations (L, luminance; C, chromatic; LC, luminance and chromatic); both online and offline performances were evaluated. RESULTS: The accuracy rate under the online LC condition was 80.6%. Offline analysis showed that the LC condition was associated with significantly higher accuracy than was the L or C condition (Tukey-Kramer, p < 0.05). No significant difference was observed between L and C conditions. CONCLUSIONS: The LC condition, which used the green/blue flicker matrix was associated with better performances in the P300 BCI. SIGNIFICANCE: The green/blue chromatic flicker matrix can be an efficient tool for practical BCI application. PMID- 19560966 TI - Development of a PCR assay for typing and subtyping of Brucella species. AB - In the course of this study, examinations were carried out to develop a PCR-based test which allows discrimination of Brucella species and biovars not targeted by the currently established gel-based PCR assays. Appropriate primers were designed based on specific deletions and insertions in the different Brucella genomes as determined by RAPD-PCR and whole-genome comparisons. After testing the specificity of the primers with a set of 22 Brucella reference strains of all species and biovars, they were used to supplement the existing PCR assays resulting in a 19-primer multiplex PCR. In addition to the commonly used PCR assays, the developed assay specifically identified B. neotomae, B. pinnipedialis, B. ceti, and B. microti. Furthermore, it differentiated B. abortus biovars 1, 2, 4 from biovars 3, 5, 6, 9, as well as between B. suis biovar 1, biovars 3, 4, and biovars 2 and 5. When tested in the multiplex assay, all Brucella type and reference strains and the majority of 118 field strains examined could be accurately identified by their respective banding patterns according to their previous typing. B. canis strains were subdivided into 2 groups, one exhibiting a unique pattern and the other one a banding pattern shared with B. suis biovars 3 and 4. Species of the closely related genus Ochrobactrum and several other clinically relevant bacteria showed no amplification product. Hence, the developed PCR assay is useful for rapid identification of Brucella at the species and at the biovar level. PMID- 19560967 TI - Modelling the black death. A historical case study and implications for the epidemiology of bubonic plague. AB - We analysed a plague outbreak in the mining town of Freiberg in Saxony which started in May 1613 and ended in February 1614. This epidemic was selected for study because of the high quality of contemporary sources. It was possible to identify 1400 individual victims meaning that more than 10% of the population of the city perished. The outbreak was modelled by 9 differential equations describing flea, rat, and human populations. This resulted in a close fit to the historical records of this outbreak. An interesting implication of the model is that the introduction of even a small number of immune rats into an otherwise unchanged setting results in an abortive outbreak with very few human victims. Hence, the percentage of immune rats directly influences the magnitude of a human epidemic by diverting search activities of the fleas. Thus, we conclude that the spread of Rattus norvegicus, which might acquire partial herd immunity by exposure to soil- or water-borne Yersinia species due to its preference for wet habitats, contributed to the disappearance of Black Death epidemics from Europe in the 18th century. In order to prove whether or not the parameter values obtained by fitting a given outbreak are also applicable to other cases, we modelled the plague outbreak in Bombay 1905/06 using the same parameter values except for the number of humans as well as of immune and susceptible rats. PMID- 19560968 TI - Prediction of VO2max from a new field test based on portable indirect calorimetry. AB - We assessed the validity and reliability of the new 15m square shuttle run test (SST) for predicting laboratory treadmill test (TT) maximal oxygen uptake (VO(2 max)) compared to the 20 m multistage shuttle run test (MST) in 45 adult males. Thirty participants performed a TT and a SST once to develop a VO( 2max) prediction model. The remaining 15 participants performed the TT and MST once and the SST twice for cross-validation purposes. Throughout testing V O(2max) was determined via portable indirect calorimetry while blood lactate concentration was assessed at the fifth recovery minute. Comparisons of TT V O(2 max) (51.3+/ 3.1 ml kg(-1)min(-1)) with SST measured (51.2+/-3.2 ml kg(-1)min(-1)) and predicted (50.9+/-3.3 ml kg(-1)min(-1)) V O(2 max) showed no differences while TT blood lactate was higher compared to SST (10.3+/-1.7 mmol vs. 9.7+/-1.7 mmol, respectively). In contrast, MST measured (53.4+/-3.5 ml kg(-1)min(-1)) and predicted (57.0+/-4.5 ml kg(-1)min(-1)) V O(2 max) and blood lactate (11.2+/-2.0 mmol) were significantly higher compared to TT. No test-retest differences were detected for SST measured and predicted V O(2 max) and blood lactate. It is concluded that the SST is a highly valid and reliable predictive test for V O(2 max). PMID- 19560969 TI - Work-rate of substitutes in elite soccer: a preliminary study. AB - The aim of this study was to investigate the work-rate of substitutes in professional soccer. A computerised player tracking system was used to assess the work-rates of second-half substitutes (11 midfielders and 14 forwards) in a French Ligue 1 club. Total distance, distance covered in five categories of movement intensity and recovery time between high-intensity efforts were evaluated. First- and second-half work-rates of the replaced players were compared. The performance of substitutes was compared to that of the players they replaced, to team-mates in the same position who remained on the pitch after the substitution and in relation to their habitual performances when starting games. No differences in work-rate between first- and second-halves were observed in all players who were substituted. In the second-half, a non-significant trend was observed in midfield substitutes who covered greater distances than the player they replaced whereas no differences were observed in forwards. Midfield substitutes covered a greater overall distance and distance at high-intensities (p<0.01) and had a lower recovery time between high-intensity efforts (p<0.01) compared to other midfield team-mates who remained on the pitch. Forwards covered less distance (p<0.01) in their first 10-min as a substitute compared to their habitual work-rate profile in the opening 10-min when starting matches while this finding was not observed in midfielders. These findings suggest that compared to midfield substitutes, forward substitutes did not utilise their full physical potential. Further investigation is warranted into the reasons behind this finding in order to optimise the work-rate contributions of forward substitutes. PMID- 19560970 TI - Counting organised sport injury cases: evidence of incomplete capture from routine hospital collections. AB - Organised sports are a popular form of physical activity, but unfortunately, participation can result in injury. Despite this, there have been surprisingly few studies that have reported the population rate of sports injury. Data from the 2005 New South Wales (NSW, Australia) Population Health Survey were analysed to describe self-reported injury experiences during participation in organised sports activities and the source of treatment for such injuries during a 12-month period in a population representative sample of adults aged 16+ years. At interview, 2414 respondents stated that they had participated in organised sport in the previous 12 months and just under one-third (30.9%) reported that they had been injured during this participation. Half of all injuries required formal treatment from a health or medical practitioner. Physiotherapists most commonly provided treatment for sports injury (26.6% of cases) followed by general practitioners (15.6%). Only 2.8% of all injured sports participants were admitted to hospital for their injury and a further 6.1% received treatment in an emergency department. This corresponds to at most only 8.9% of all treated sports injuries receiving treatment in a hospital setting. Population-based estimates of the rate and burden of sports injuries that rely solely on routine hospital data collections are likely to grossly underestimate the size of the problem, as very few cases are treated in a hospital setting. PMID- 19560971 TI - Normative values for three clinical measures of motor performance used in the neurological assessment of sports concussion. AB - Postural control and motor coordination are essential components of normal athletic activity. Tasks involving balance and coordination are used to determine neurological function in sports-related concussion. Determining normative values for these tasks is therefore essential to provide sports medicine professionals with a frame of reference with which to interpret clinical measures obtained from players suspected of sustaining a concussion. One hundred and seventytwo healthy subjects (16-37 yrs) performed three timed tests: Tandem Gait (TG); Finger-to Nose (FTN); Single-Leg-Stance (SLS) on firm and foam surfaces. Unadjusted geometric means (+/-SD) for each measure were averaged across three trials. Time to complete TG was 11.2+/-1.2s. FTN for the dominant and non-dominant arm were 2.9+/-1.1s and 3.0+/-1.2s, respectively. SLS values for dominant and non-dominant leg were 20.4+/-3.0s (firm), 3.4+/-1.6s (foam), and 21.0+/-2.9s (firm), 3.3+/ 1.6s (foam), respectively. For TG, there was an order effect (P<.001) but no age, sex or BMI effects. FTN demonstrated a dominant arm preference (P<.001), sex (P=.006), BMI (P=.043) and order effects (P<.001). SLS demonstrated an order effect on the firm surface (P=.009) and an order (P<.001) and BMI (P=.001) effect on foam. Intra-rater reliability, as measured by ICC (3,3), demonstrated that TG and FTN had excellent reliability compared to SLS. FTN and TG should continue to be used in test batteries to determine neurological function in sports-related concussion. PMID- 19560972 TI - Effects of active vs. passive recovery on repeated rugby-specific exercises. AB - The aim of this study was to determine the effects of active vs. passive recovery on performance of a rugby-specific intermittent test in rugby union players. Seven male rugby players (20.6+/-0.5 yrs; 181.9+/-10.0 cm; 94.5+/-12.8 kg) performed in random order, over two separate sessions, a specific repeated-sprint rugby test, the Narbonne test (6 x 4 consecutive actions: 1, scrummaging; 2, agility sprinting; 3, tackling; 4, straight sprinting) with 30s of passive or active recovery (running at 50% of maximal aerobic speed). The Narbonne tests were completed before (pre-test) and after (post-test) a 30-min rugby match. During the Narbonne test, scrum forces, agility and sprint times, heart rate and rate of perceived exertion were measured. Scrum forces were lower in active (74.9+/-13.4 kg) than in passive recovery (90.4+/-20.9 kg), only during the post test (p<0.05). Fatigue index (%) (p<0.05) and total sprint time (s) (p<0.01) were significantly greater in active than in passive recovery, both during the pre test (11.5+/-5.7% vs. 6.7+/-4.5% and 18.1+/-1.3s vs. 16.9+/-0.9s) and the post test (7.3+/-3.3% vs. 4.3+/-1.5% and 18.3+/-1.6s vs. 16.9+/-1.1s). Consequently, the results indicated that passive recovery enabled better performance during the Narbonne test. However, it is obviously impractical to suggest that players should stand still during and following repeated-sprint bouts: the players have to move to ensure they have taken an optimal position. PMID- 19560973 TI - The impact of socioeconomic position on sport participation among South Australian youth. AB - Organised sport among youth makes a substantial contribution to daily energy expenditure. This study investigated socioeconomic gradients in sport participation and predictors of participation. A representative sample of young South Australians (10-15 y; n=1737) was surveyed on organised sport participation in the previous 12 months, and predictors derived from the Children's Physical Activity Correlates scale and a parent survey. Four constructs were derived: 'is it worth it?' (perceived outcomes); 'am I able?' (perceived competency); 'reinforcing' (parental support); and 'enabling' (perceived barriers, from the parent survey). Socioeconomic position (SEP) was operationalized by an area-level indicator, the Socioeconomic Indicator for Advantage (SEIFA), split into tertiles. Sport participation was higher among high (highest SEIFA tertile) compared with low (lowest SEIFA tertile) SEP children. All predictors except 'am I able?' were positively associated with sport participation among boys and girls. Of these predictors, the 'enabling' construct varied by SEP among both boys and girls, with high SEP children reporting fewer barriers to participation. High SEP girls reported higher scores on 'reinforcing' and 'is it worth it?' than their low SEP counterparts. Low SEP girls reported lower levels of both instrumental and affective support from parents to play sport. There are distinct SEP gradients in sport participation, as well as its psychosocial and environmental predictors among South Australian youth. Low SEP girls are the most disadvantaged in terms of parental support to participate in sport. Interventions targeting this vulnerable group are urgently required. PMID- 19560974 TI - Organelle interplay in peroxisomal disorders. AB - Peroxisomes are no longer regarded as autonomous organelles because evidence for their interplay with other cellular organelles is emerging. Peroxisomes interact with mitochondria in several metabolic pathways, including beta-oxidation of fatty acids and the metabolism of reactive oxygen species. Both organelles are in close contact with the endoplasmic reticulum (ER) and share several proteins, including organelle fission factors. Today, the study of peroxisome biogenesis disorders mainly focuses on metabolic defects such as accumulation of very long chain fatty acids or plasmalogen deficiency. In addition to metabolic dysregulation, mitochondria and ER abnormalities have also been observed. Whether these contribute to disease pathology is not yet known, but recent findings suggest that this possibility should be considered. Here, we discuss the potential involvement of organelle interplay in peroxisomal disorders. PMID- 19560975 TI - Anticonvulsant, anxiolytic, and sedative properties of the roots of Nauclea latifolia Smith in mice. AB - Root bark of Nauclea latifolia Smith (Rubiaceae) was evaluated for its anticonvulsant, anxiolytic, and sedative activity in mice. Animal models (maximal electroshock-, pentylenetetrazol-, and strychnine-induced convulsions; N-methyl-D aspartate-induced turning behavior; elevated plus maze; stress-induced hyperthermia; open field; and diazepam-induced sleep) were used. The decoction from the bark of the roots of N. latifolia strongly increased the total sleep time induced by diazepam. It also protected mice against maximal electroshock-, pentylenetetrazol-, and strychnine-induced seizures. In addition, turning behavior induced by N-methyl-D-aspartate was inhibited. N. latifolia antagonized, in a dose-dependent manner, stress-induced hyperthermia and reduced body temperature. In the elevated plus maze, N. latifolia increased the number of entries into, percentage of entries into, and percentage of time in open arms, and reduced rearing, head dipping, and percentage of time in closed arms. In the open field test, N. latifolia increased crossing and reduced rearing and defecation. It could be concluded that the decoction of N. latifolia, used in traditional medicine in Cameroon in the treatment of fever, malaria, insomnia, anxiety and epilepsy seemed to possess, sedative, anticonvulsant, anxiolytic and antipyretic properties in mice. PMID- 19560976 TI - Sex differences in prior pain experience. AB - Sex differences in clinical and experimental pain experiences are well documented. However, there has been little work investigating men's and women's experiences with common painful events. This study examined sex differences in the nature and intensity of common pain experiences. Participants (102 women and 85 men) completed the Prior Pain Experience Questionnaire, which is a 79-item assessment of an individual's pain experience, recalled pain ratings, and imagined pain ratings. Analyses of variance were conducted to assess for sex differences in overall pain experience and pain ratings. Men and women did not have significant differences in the overall number of reported pain experiences or in the overall mean pain rating of those experiences. However, they differed in specific pain events experienced (eg, men experienced concussions more than women) and pain ratings (eg, women rated minor surgery as significantly more painful than men). Individuals who imagined pain events tended to rate them as equally or more painful than individuals who experienced those pain events. PERSPECTIVE: Results of this study demonstrate that men and women have varying types of pain experiences without evidence that the overall pain experience differs between sexes. It was also found that imagined pain ratings are often worse than experienced pain ratings, lending support for the tendency of individuals to catastrophize. PMID- 19560977 TI - Life-threatening postoperative hypoventilation after bariatric surgery. PMID- 19560978 TI - Correlations between intra-abdominal pressure and obesity-related co-morbidities. AB - BACKGROUND: Obesity is associated with chronic increases in intra-abdominal pressure (IAP). The aim of the present study was to examine the correlation between the IAP and the number of obesity-related co-morbidities. METHODS: A total of 63 morbidly obese patients who were undergoing bariatric surgery had their IAP measured intraoperatively while in a supine position and under general anesthesia. The IAP readings were obtained through an indwelling urinary bladder catheter. The correlation of obesity-related co-morbidities, including systemic hypertension, type 2 diabetes mellitus, gastroesophageal reflux disease, urinary stress incontinence, lower extremity edema, obstructive sleep apnea, and abdominal wall hernia, and the level of IAP were examined using a stepwise regression analysis model. RESULTS: Of the 62 patients, 57 were women. The mean age was 44 + or - 11 years, and the body mass index was 49 + or - 10 kg/m(2). Of the 62 patients, 48 (77%) had an elevated IAP (> or = 9 cm H(2)O). A significant and positive correlation was found between the IAP level and the number of obesity-related co-morbidities (Pearson's r = .8; P <.05). Stepwise logistic regression analysis revealed that systemic hypertension, American Society of Anesthesiologists score, and body mass index were predictors of elevated IAP. A normal IAP appeared to offer a protective effect against systemic hypertension. CONCLUSION: In this cohort of mainly obese women, the baseline IAP of morbidly obese patients was abnormally elevated. A greater IAP correlated with the presence of a greater number of obesity-related co-morbid conditions. Systemic hypertension was significantly associated with an elevated IAP. Chronic increases in IAP might, in part, be responsible for the pathogenesis of systemic hypertension in the morbidly obese. PMID- 19560980 TI - Laparoscopic single-site surgery for placement of adjustable gastric band--a series of 22 cases. AB - BACKGROUND: We present a series of 22 patients who underwent laparoendoscopic single-site (LESS) surgery for placement of an adjustable gastric band at a U.S. university hospital. METHODS: From December 2007 to December 2008, LESS surgery, through a transumbilical incision, to place an adjustable gastric band was performed on 22 patients under institutional review board approval. Multiple ports were placed through a single incision in the umbilicus to allow for liver retraction, visualization, and the working instruments. None of the critical steps of the standard pars flaccida technique were altered. RESULTS: A total of 22 patients were carefully selected and included 20 women and 2 men, with an age range of 18-67 years (mean 42). The mean body mass index was 42 kg/m(2) (range 35 45). The exclusion criteria included hepatomegaly, central obesity, previous abdominal surgery, and super-obesity. The mean operative time was 84 minutes (range 53-111). All patients were discharged home within the 23-hour admission, and no perioperative complications were noted. In addition, no wound-related complications developed. One patient required conversion to conventional laparoscopy. No intraoperative or postoperative complications occurred. CONCLUSION: In our experience, LESS surgery for adjustable gastric band placement shows this technique to be both feasible and safe in selected patients to date. Although technical limitations exist that will be improved on, additional studies are needed to compare LESS surgery for placement of an adjustable gastric band with traditional laparoscopic techniques. PMID- 19560979 TI - Peroral endoscopic anastomotic reduction improves intractable dumping syndrome in Roux-en-Y gastric bypass patients. AB - BACKGROUND: Dumping syndrome is a well-described consequence of Roux-en-Y gastric bypass. Although the condition can benefit some patients with morbid obesity, a subset will develop intractable dumping syndrome characterized by symptomatic episodes with most meals. We describe the first series of patients successfully treated endoscopically for intractable dumping syndrome. METHODS: Endoscopic gastrojejunal anastomotic reduction was performed in patients with intractable dumping syndrome after Roux-en-Y gastric bypass using a combination of argon plasma coagulation, endoscopic suturing, and fibrin glue. The technical feasibility of endoscopic anastomotic reduction and the clinical improvement in dumping symptoms were assessed by clinical follow-up. RESULTS: Endoscopic anastomotic reduction was technically successful in 6 consecutive patients with a dilated gastrojejunal anastomosis and intractable dumping syndrome. One patient reported hematemesis 2 days after the procedure that was treated endoscopically. No other significant complications occurred. Complete and persistent resolution of the dumping symptoms was achieved in all patients, with a median follow-up of 636 days. CONCLUSION: Endoscopic anastomotic reduction appears technically feasible and safe and might be a minimally invasive treatment option for patients who experience intractable dumping symptoms after Roux-en-Y gastric bypass. Additional studies are needed to determine the long-term efficacy of this procedure. PMID- 19560981 TI - One size does not fit all: CMS payment suspensions do not consider the unique risks and challenges of the morbidly obese. PMID- 19560982 TI - Bariatric surgery and hypertension. PMID- 19560984 TI - Micronutrient-responsive cerebral dysfunction other than Wernicke's encephalopathy after malabsorptive surgery. AB - BACKGROUND: Micronutrient-responsive cerebral dysfunction (MRCD) is known to occur after gastric restrictive and malabsorptive weight loss surgery. Wernicke's encephalopathy (WE) is often considered the etiology, but many cases without thiamine deficiency have been reported. It is important to recognize that other micronutrient-responsive cerebral dysfunctions exist in this high-risk group. We evaluated the published data for evidence of MRCD after gastric restrictive and malabsorptive weight loss surgery. METHODS: A search of PubMed and OVID was conducted to identify all published cases of cerebral dysfunction after gastric restrictive and malabsorptive weight loss surgery. The cases were compared using the presenting clinical signs and symptoms and objective confirmatory findings. The cases were stratified according to the type of surgery and the known or suspected etiology of the cerebral dysfunction. RESULTS: A total of 65 cases of cerebral dysfunction after gastric restrictive and/or malabsorptive weight loss surgery were identified. A careful analysis of all cases revealed that 48 (73.8%) could be attributed to WE. The remaining cases represented a subset with a pattern of cerebral dysfunction incompatible with WE. This condition most likely resulted from multiple micronutrient deficiencies (MRCD). Patients with MRCD were more likely to have undergone a malabsorptive surgical procedure (P = .001), to present with dysarthria (P <.001), and less likely to have ophthalmoplegia (P <.001) or nystagmus (P = .02) compared with those with WE. CONCLUSION: Current evidence supports the identification of MRCD after malabsorptive weight loss surgery that is not compatible with WE. Clinicians should be aware of this clinical entity and be prepared to treat it appropriately. PMID- 19560983 TI - Comparison of weight loss and body composition changes with four surgical procedures. AB - BACKGROUND: A paucity of information is available on the comparative body composition changes after bariatric procedures. The present study reports on the body mass index (BMI) and body composition changes after 4 procedures by a single group. METHODS: At the initial consultation, the weight and body composition of the patients undergoing 4 different bariatric procedures were measured by bioimpedance (Tanita 310). Follow-up examinations were performed at 1 year and at subsequent visits after surgery. Analysis of variance was used to compare the postprocedure BMI and body composition. Analysis of covariance was used to adjust for baseline differences. RESULTS: A total of 101 gastric bypass (GB) patients were evaluated at 19.1 + or - 10.6 months, 49 biliopancreatic diversion with the duodenal switch (BPD/DS) patients at 27.5 + or - 16.3 months, 41 adjustable gastric band (AGB) patients at 21.4 + or - 9.2 months, and 30 sleeve gastrectomy (SG) patients at 16.7 + or - 5.6 months (P <.0001). No differences were found in patient age or gender among the 4 groups. The mean preoperative BMI was significantly different among the 4 groups (P <.0001): 61.4 kg/m(2), 53.2, 46.7, and 44.3 kg/m(2) for the SG, BPD/DS, GB, and AGB group, respectively. The postoperative BMI adjusted for baseline differences was 27.8 (difference 23.6 + or - 8.3), 32.5 (difference 15.6 + or - 5.0), 37.2 (difference 18.2 + or - 8.2), and 39.5 kg/m(2) (difference 7.5 + or - 4.3) for the BPD/DS, GB, SG, and AGB groups, respectively (P <.0001). The percentage of excess weight loss was 84%, 70%, 49%, and 38% for the BPD/DS, GB, SG, and AGB groups, respectively (P <.0001). The postoperative percentage of body fat adjusted for baseline differences was 25.7% (23.9% + or - 7.0%) 32.7% (16.1% + or - 10.5%) 37.7% (16.7% + or - 5.6%), and 42% (6.0% + or - 6.8%) for the BPD/DS, GB, SG, and AGB groups, respectively (P <.0001). The lean body mass changes were reciprocal. CONCLUSION: Although the BPD/DS procedure reduced the BMI the most effectively and promoted fat loss, all the procedures produced weight loss. The AGB procedure resulted in less body fat loss within 21.5 months than SG within 16.7 months. Longer term observation is indicated. PMID- 19560985 TI - Comparative determination of methyl mercury in whole blood samples using GC-ICP MS and GC-MS techniques. AB - Two methods for the determination of methyl mercury (MeHg) in whole blood samples based on different mass spectrometric detection techniques are compared. The methods were employed in two studies in which the internal exposure of a group of mercury-exposed workers to total mercury and MeHg was investigated. Blood samples of these workers were analysed for MeHg independently from each other in two laboratories using similar extraction procedures but different detection techniques, viz. coupled GC-EI-MS/ICP-MS and GC-MS using D(3)-MeHg as internal standard. MeHg was detected in all blood samples in concentrations ranging from 0.3 to 9.0 microg/L. Though different detection techniques were employed, the results obtained by the two laboratories were in relatively good agreement. PMID- 19560986 TI - Analysis of neurosterols by GC-MS and LC-MS/MS. AB - The term neurosteroid was coined by Baulieu and colleagues in Paris towards the end of the last century to describe steroids which are synthesised in the central or peripheral nervous system [E.E. Baulieu, Psychoneuroendocrinology 23 (1998) 963-87]. This definition was restricted to side-chain "shortened" steroids with 21 carbon atoms or less, and excluded sterols and their carboxylic acids with an intact side-chain. By analogy, we now use the term neurosterol to describe C(27) sterols synthesised in the nervous system. In this review we discuss the biological importance of neurosterols, and how they are extracted, isolated, and analysed by GC-MS and LC-MS/MS, from brain and relevant body fluids. We present applications of methodology employed for analysis of specific sterols and comment on the relative merits of the methods employed. Finally, the importance of future in-depth "sterolomic" investigations of brain is highlighted. PMID- 19560987 TI - Determination of glutathione and glutathione disulfide in biological samples: an in-depth review. AB - Glutathione (GSH) is a thiol-containing tripeptide, which plays central roles in the defence against oxidative damage and in signaling pathways. Upon oxidation, GSH is transformed to glutathione disulfide (GSSG). The concentrations of GSH and GSSG and their molar ratio are indicators of cell functionality and oxidative stress. Assessment of redox homeostasis in various clinical states and medical applications for restoration of the glutathione status are of growing importance. This review is intended to provide a state-of-the-art overview of issues relating to sample pretreatment and choices for the separation and detection of GSH and GSSG. High-performance liquid chromatography, capillary electrophoresis and gas chromatography (as techniques with a separation step) with photometric, fluorimetric, electrochemical and mass spectrometric detection are discussed, stress being laid on novel approaches. PMID- 19560988 TI - Childhood overweight in the United States: a quantile regression approach. AB - The prevalence of overweight children in the United States has increased dramatically over the past two decades, and is creating well-known public health problems. Moreover, there is also evidence that children who are not overweight are becoming heavier. We use quantile regression models along with standard ordinary least squares (OLS) models to explore the correlates of childhood weight status and overweight as measured by the Body Mass Index (BMI). This approach allows the effects of covariates to vary depending on where in the BMI distribution a child is located. Our results indicate that OLS masks some of the important correlates of child BMI at the upper and lower tails of the weight distribution. For example, mother's education has no effect on black children, but is associated with improvements in BMI for overweight white boys and underweight white girls. Conversely, mother's cognitive aptitude has no effect on white boys, but is associated with BMI improvements for underweight black children and overweight white girls. Further, we find that underweight white children and black girls experience similar improvements in BMI as they get older, but that for black boys there is little if any association between age and BMI anywhere in the BMI distribution. PMID- 19560989 TI - Urbanization and the spread of diseases of affluence in China. AB - We quantify, track and explain the distribution of overweight and of hypertension across Chinese provinces differentiated by their degree of urbanicity over the period 1991-2004. We construct an index of urbanicity from longitudinal data on community characteristics from the China Health and Nutrition Survey and compute, for the first time, a rank-based measure of inequality in disease risk factors by degree of urbanicity. Prevalence rates of overweight and hypertension almost doubled between 1991 and 2004 and these disease risk factors became less concentrated in more urbanized areas. Decomposition analysis reveals that one half of the urbanicity-related inequality in overweight is directly attributable to community level characteristics, while for hypertension the contribution of such characteristics increased from 20% in 1991 to 62% in 2004. At the individual level, lower engagement in physical activity and farming explain more than half of the urban concentration of overweight and a rising share (28%) of the greater prevalence of hypertension in more urbanized areas. Higher incomes explain around one-tenth of the urban concentration of both overweight and hypertension, while the education advantage of urban populations has a similar sized offsetting effect. PMID- 19560991 TI - Improvement of mouse cardiac function by hESC-derived cardiomyocytes correlates with vascularity but not graft size. AB - Transplantation of human embryonic stem cell-derived cardiomyocytes (hESC-CM) has been shown to improve the function of the rodent heart 1 month after myocardial infarction (MI). However, the mechanistic basis and optimal delivery strategies are unclear. We investigated the influence of the number of injected cells, resulting graft size, and possible paracrine mechanisms in this process. MI was induced in NOD-SCID mice (n=84) followed by injection of enriched hESC-CM at different dosages, hESC-non-CM derivatives, culture medium, or no injection. Cardiac function was monitored for 12 weeks with 9.4 T MRI (n=70). Grafts were identified by epifluorescence of a transgenic GFP marker and characterized by immunofluorescence. Vascularity and paracrine effects were investigated immunohistochemically. Transplantation of differentiated hESCs improved short, mid-, and long-term cardiac performance and survival, although only cardiomyocytes formed grafts. A mid-term (4 weeks) cardiomyocyte-specific enhancement was associated with elevated vascular density around the graft and attenuated compensatory remodeling. However, increasing the number of hESC-CM for injection did not enhance heart function further. Moreover, we observed that small graft size was associated with a better functional outcome. HESC-CM increased myocardial vascularization and enhanced heart function in mice after MI, but larger graft size was associated with reduced functional improvement. Future studies should focus on advanced delivery strategies and mechanisms of action rather than increasing graft size. PMID- 19560990 TI - At the crossroads of polarity, proliferation and apoptosis: the use of Drosophila to unravel the multifaceted role of endocytosis in tumor suppression. AB - Endocytosis is an important regulator of cell-cell signaling and endocytic trafficking has been increasingly implicated in control of tumor suppression. Recent insights from Drosophila indicate that impairment of multiple trafficking steps which lead to receptor degradation can cause tumor formation in epithelial organs. These tumors are characterized by sustained activation of a number of mitogenic signaling pathways, and by subversion of epithelial polarity and the apoptotic response. Cooperation between such alterations, as well as tumor-host interactions, is also observed. The recapitulation of several hallmarks of human cancers in fly tumors provides a framework to understand the role of defective endocytosis in cancer. PMID- 19560992 TI - Childhood obesity and neighborhood food-store availability in an inner-city community. AB - OBJECTIVE: Prior studies have shown an association between fast-food restaurants and adolescent body size. Less is known about the influence of neighborhood food stores on a child's body size. We hypothesized that in the inner-city, minority community of East Harlem, New York, the presence of convenience stores and fast food restaurants near a child's home is associated with increased risk for childhood obesity as measured by body mass index (BMI). DESIGN: Baseline data of 6- to 8-year-old East Harlem boys and girls (N=323) were used. Anthropometry (height and weight) was conducted with a standardized protocol. Food-store data were collected via a walking survey. Stores located within the same census block as the child's home address were identified by using ArcGIS 8.3. We computed age- and sex-specific BMI percentiles by using national norms of the Centers for Disease Control and Prevention. Using odds ratios, we estimated risk of a child's BMI percentile being in the top tertile based on number and types of food stores on their census blocks. RESULTS: Convenience stores were present in 55% of the surveyed blocks in which a study particpant lived and fast-food restaurants were present in 41%. Children (n=177) living on a block with 1 or more convenience stores (range, 1-6) were more likely to have a BMI percentile in the top tertile (odds ratio 1.90, 95% confidence interval, 1.15-3.15) compared with children having no convenience stores (n=146). CONCLUSIONS: The presence of convenience stores near a child's residence was associated with a higher BMI percentile. This has potential implications for both child- and neighborhood-level childhood obesity interventions. PMID- 19560993 TI - Prevalence and trends of severe obesity among US children and adolescents. AB - OBJECTIVE: To determine the extent to which the 2007 definitions for severe obesity (body mass index [BMI] > or = 99th percentile for age and gender) and morbid obesity (BMI > or = 40kg/m(2)) affects different groups of American children and adolescents and has increased over time. METHODS: Analysis of nationally representative data from the National Health and Nutrition Examination Survey (NHANES) II, III, and 1999-2004; 12 384 US children and adolescents ages 2 to 19 years were included in the analysis. Outcome measures were the proportion of subjects with severe and morbid obesity, with age, gender, race, and poverty income ratio (PIR) as key variables. RESULTS: In 1999-2004, 3.8% of children 2 to 19 years old had a BMI in the > or = 99th percentile, with higher prevalence among boys than girls (4.6% vs 2.9%; P < .001). Prevalence was highest among blacks, 5.7% and Mexican Americans, 5.2%, compared with whites, 3.1% (P < .001). The prevalence differed by the PIR category as well (4.3% for those with PIR < or = 3 vs 2.5% for those with PIR>3; P=.002). BMI > or = 40kg/m(2) was found in 1.3% of adolescents ages 12 to 19 years, with similar associations with race and poverty. The overall prevalence of BMI > or = 99th percentile has increased by more than 300% since NHANES II (1976), and over 70% since NHANES III (1994) in children 2 to 19 years of age. CONCLUSIONS: Rates of severe childhood obesity have tripled in the last 25 years, with significant differences by race, gender, and poverty. This places demands on health care and community services, especially because the highest rates are among children who are frequently underserved by the health care system. PMID- 19560995 TI - Effects of different stretching techniques on the outcomes of isokinetic exercise in patients with knee osteoarthritis. AB - We recruited 132 subjects with bilateral knee osteoarthritis (Altman Grade II) to compare the effects of different stretching techniques on the outcomes of isokinetic muscle strengthening exercises. Patients were randomly divided into four groups (I-IV). The patients in Group I received isokinetic muscular strengthening exercises, Group II received bilateral knee static stretching and isokinetic exercises, Group III received proprioceptive neuromuscular facilitation (PNF) stretching and isokinetic exercises, and Group IV acted as controls. Outcomes were measured by changes in Lequesne's index, range of knee motion, visual analog pain scale, and peak muscle torques during knee flexion and extension. Patients in all the treated groups experienced significant reductions in knee pain and disability, and increased peak muscle torques after treatment and at follow-up. However, only patients in Groups II and III had significant improvements in range of motion and muscle strength gain during 60 degrees/second angular velocity peak torques. Group III demonstrated the greatest increase in muscle strength gain during 180 degrees/second angular velocity peak torques. In conclusion, stretching therapy could increase the effectiveness of isokinetic exercise in terms of functional improvement in patients with knee osteoarthritis. PNF techniques were more effective than static stretching. PMID- 19560994 TI - Hepatitis C virus infection and metabolic syndrome---a community-based study in an endemic area of Taiwan. AB - Metabolic syndrome (MS) is a complicated disorder associated with a high risk of future development of micro- and macrovascular complications. The extrahepatic manifestations of hepatitis C virus (HCV) infection can include multiple metabolic abnormalities. However, the extent, severity, and characteristics of MS in HCV-infected patients have rarely been investigated in community-based settings. This study aimed to determine the difference in prevalence and distribution of the components of MS between HCV-infected patients and healthy controls. Multipurpose mass screening of adults was conducted in an HCV-endemic area of Southern Taiwan. Clinical profiles in terms of anthropometric data and MS components, as well as viral hepatitis markers, were assessed. Two hundred and thirty-seven adults (94 males; mean age, 55.5 +/- 10.8 years) were recruited. The prevalence of anti-HCV seropositivity was 39.2% (93/237). The prevalence of MS was higher in the HCV-infected individuals (24.7%, 23/93) than in the control, uninfected subjects (13.2%, 19/144, p = 0.02). In terms of MS components, HCV infected subjects had a higher prevalence of high waist circumference (51.6% vs. 25.7%, p < 0.001) and hypertension (58.1% vs. 36.8%, p = 0.001) than controls. Multivariate logistic regression analysis demonstrated that anti-HCV positivity was significantly associated with MS (odds ratio, 6.4; 95% confidence interval, 1.82-22.84; p = 0.004). HCV infection was associated with a higher prevalence of MS. Determination of MS in patients with HCV infection could therefore be indicated. PMID- 19560996 TI - Adverse situations encountered by adolescent students who return to school following suspension. AB - This study aimed to investigate the adverse personal, family, peer and school situations encountered by adolescent students who had returned to school after being suspended. This was a large-scale study involving a representative population of Taiwanese adolescents. A total of 8,494 adolescent students in Southern Taiwan were recruited in the study and completed the questionnaires. The relationships between their experiences of suspension from school and adverse personal, family, peer, and school situations were examined. The results indicated that 178 (2.1%) participants had been suspended from school at some time. Compared with students who had never been suspended, those who had experienced suspension were more likely to report depression, low self-esteem, insomnia, alcohol consumption, illicit drug use, low family support, low family monitoring, high family conflict, habitual alcohol consumption, illicit drug use by family members, low rank and decreased satisfaction in their peer group, having peers with substance use and deviant behaviors, low connectedness to school, and poor academic achievement. These results indicate that adolescent students who have returned to school after suspension encounter numerous adverse situations. The psychological conditions and social contexts of these individuals need to be understood in depth, and intervention programs should be developed to help them to adjust when they return to school and to prevent school dropouts in the future. PMID- 19560998 TI - Urethral stripping caused by stent removal and its successful treatment with hyperbaric oxygen therapy: a case report. AB - Urethral stents usually provide initial, dramatic relief from obstructive voiding symptoms. However, complications such as recurrent urinary tract infections, stent migration, encrustation and recurrent urethral strictures are not rare, and stents should be removed when complications occur. Urethral stent removal is associated with potential risks of urethral injury, bleeding and external sphincter trauma. Hyperbaric oxygen therapy (HBO) accelerates wound healing by increasing tissue microcirculation, decreasing capillary pressure and resolving tissue edema. Although HBO has been used in various urologic applications, there is no report of HBO therapy being used to treat isolated urethral injuries. Here we present a case of urethral stripping caused by endoscopic stent removal, and its conservative treatment with HBO therapy. PMID- 19560997 TI - Measurement of the functional status of patients with different types of muscular dystrophy. AB - Muscular dystrophy (MD) comprises a group of diseases characterized by progressive muscle weakness that induces functional deterioration. Clinical management requires the use of a well-designed scale to measure patients' functional status. This study aimed to investigate the quality of the functional scales used to assess patients with different types of MD. The Brooke scale and the Vignos scale were used to grade arm and leg function, respectively. The Barthel Index was used to evaluate the function of daily living activity. We performed tests to assess the acceptability of these scales. The characteristics of the different types of MD are discussed. This was a multicenter study and included patients diagnosed with Duchenne muscular dystrophy (DMD) (classified as severely progressive MD), Becker muscular dystrophy (BMD), limb girdle muscular dystrophy (LGMD) and facioscapulohumeral muscular dystrophy (FSHD). BMD, LGMD, and FSHD were classified as slowly progressive MD. The results demonstrated that the Brooke scale was acceptable for grading arm function in DMD, but was unable to discriminate between differing levels of severity in slowly progressive MD. The floor effect was large for all types of slowly progressive MD (range, 20.0 61.9), and was especially high for BMD. The floor effect was also large for BMD (23.8%) and FSHD (50.0%) using the Vignos scale. Grades 6-8 of the Vignos scale were inapplicable because they included items involving the use of long leg braces for walking or standing, and some patients did not use long leg braces. In the Barthel Index, a ceiling effect was prominent for slowly progressive MD (58.9%), while a floor effect existed for DMD (17.9%). Among the slowly progressive MDs, FSHD patients had the best level of functioning; they had better leg function and their daily living activities were less affected than patients with other forms of slowly progressive MD. The results of this study demonstrate the acceptability of the different applications used for measuring functional status in patients with different types of MD. Some of the limitations of these measures as applied to MD should be carefully considered, especially in patients with slowly progressive MD. We suggest that these applications be used in combination with other measures, or that a complicated instrument capable of evaluating the various levels of functional status be used. PMID- 19560999 TI - Synchronous gastrointestinal stromal tumor and adenocarcinoma at the gastroesophageal junction. AB - The synchronous existence of two different tumors in the gastrointestinal tract is uncommon. We report the case of a 75-year-old man who had a concurrent gastrointestinal stromal tumor and adenocarcinoma at the gastroesophageal junction. The two tumors arose at the same site but had distinct morphologies. The etiology of synchronous tumors is still unclear and their coexistence causes problems for the surgeon, oncologist and pathologist in terms of their diagnosis, treatment, and follow-up. We report a rare case of synchronous tumors and a review of the literature. PMID- 19561000 TI - A rare occipital condyle fracture in a patient with a minor head injury. AB - Occipital condyle fracture (OCF) is an uncommon but potentially fatal disease entity. It is most commonly identified in patients suffering from severe craniocerebral trauma. The advent of computed tomography has made early detection possible. Traditional treatment using a hard neck collar is sufficient to produce solid fusion in most OCF patients. Delayed diagnosis, however, may result in neurologic deterioration due to potential displacement of fractured condylar fragments. Here we report a case of isolated, stable OCF in a patient with a minor head injury. A high level of clinical awareness of this rare disease entity is imperative for the management of traumatized patients, especially for those who have minor head injuries but persistent neck pain. PMID- 19561001 TI - Adult intussusception secondary to lymphangioma of the cecum: a case report. AB - We report the case of a patient with ileocolic intussusception caused by cecal lymphangioma. A 45-year-old man visited our hospital with a 2-month history of frequent episodes of watery diarrhea (>/= 5 times/day) and intermittent abdominal pain. A cecal submucosal tumor with mucosal ulceration and partial obstruction of the colonic lumen was identified by colonoscopy. He was admitted to our hospital 2 days later due to aggravation of his abdominal pain. Physical examination revealed tenderness over the right abdomen with no peritoneal signs. A double contrast lower gastrointestinal series showed a right-side colonic lesion with indentation and a peripheral, beak-like sign. Abdominal computed tomography scanning revealed an intra-abdominal mass with the characteristic sausage sign, highly suggestive of intussusception. The patient subsequently underwent right hemicolectomy. The final diagnosis was ileocolic intussusception with cecal lymphangioma, which was confirmed by histopathology. He had an uneventful recovery with follow-up in our hospital. This case highlights the possibility of colonic lymphangioma as the leading point of adult intussusception, and this should be taken into consideration as a possible diagnosis in this uncommon clinical condition. PMID- 19561003 TI - Maintaining bone density in patients undergoing treatment for breast cancer: is there an adjuvant benefit? AB - Women undergoing treatment for breast cancer often experience a marked decrease in bone mineral density. This decrease is observed with chemotherapy as well as endocrine therapy and is more pronounced and rapid than normal postmenopausal bone loss. Pharmacologic intervention is, therefore, necessary in many cases to preserve bone health and prevent fractures. Many small studies have demonstrated that cancer therapy-induced bone loss (CTIBL) is effectively prevented by bone targeted therapies, such as bisphosphonates and other inhibitors of bone resorption. Recently, several trials have confirmed the efficacy of bisphosphonates in the prevention of CTIBL in both premenopausal and postmenopausal women with early-stage breast cancer. In addition, concomitant treatment with zoledronic acid 4 mg every 6 months and standard adjuvant endocrine therapy has been reported to significantly improve disease-free survival and decrease disease recurrence in bone as well as other sites compared with standard therapy alone. Zoledronic acid treatment has also decreased residual tumor volume in the neoadjuvant setting. Furthermore, long-term follow up of a single study in patients with bone marrow micrometastases from breast cancer revealed overall survival benefits for patients receiving clodronate 1600 mg/day compared with placebo; however, combined results from several trials of clodronate are inconclusive. Overall, a large body of evidence is accumulating to support the potential adjuvant benefits of bisphosphonates in the treatment of early-stage breast cancer. Results from ongoing studies are expected to further elucidate the benefits of bisphosphonates in maintaining bone health and improving clinical outcomes in patients with breast cancer. PMID- 19561004 TI - Enhancing the efficacy of hormonal agents with selected targeted agents. AB - Several selected targeted agents are being investigated in combination with endocrine therapy for patients with breast cancer in an attempt to overcome or prevent endocrine resistance. The role of type I growth factor receptors epidermal growth factor receptor (EGFR) and HER2 in cross-talk with estrogen receptor (ER) signaling has been confirmed in preclinical studies in which various inhibitors have yielded additive or synergistic effects when combined with endocrine agents. Recently, several results from clinical trials investigating this concept have been reported. In ER-positive/HER-positive advanced breast cancer, the addition of trastuzumab to the aromatase inhibitor anastrozole, or the tyrosine kinase inhibitor (TKI) lapatinib to letrozole, both have significantly improved progression-free survival (PFS). The EGFR TKI gefitinib combined with tamoxifen as first-line therapy for ER-positive metastatic disease improved PFS (but not objective response rate) for patients with no previous endocrine therapy or completion of previous adjuvant therapy. A second study in a similar setting showed significant improvement in PFS for gefitinib plus anastrozole. Although it is encouraging that this approach could delay resistance, only a small proportion of patients benefit. Attempts to identify likely responders have been made in the neoadjuvant setting, with pre- and post-treatment biopsies being used to study biomarker changes. A recent preoperative study of letrozole with or without the mammalian target of rapamycin (mTOR) inhibitor everolimus reported greater tumor shrinkage for the combination, with changes in proliferation being predictive for response together with strong expression of protein S6 kinase, a downstream marker of activated mTOR. Key aspects that need to be addressed in future trials include understanding the mechanisms of action for each novel agent, designing the best trial and endpoints to demonstrate added benefit, and ensuring appropriately stratified populations based on previous endocrine exposure and/or sensitivity. PMID- 19561005 TI - A question of duration: do patients with early-stage breast cancer need more than five years of adjuvant endocrine therapy? AB - Women with hormone receptor-positive breast cancer continue to be at risk for recurrence and mortality for many years after diagnosis. Previous clinical trials established 5 years of endocrine therapy as a standard of care for both premenopausal and postmenopausal women, resulting in long-lasting benefit over shorter durations of treatment. Until recently, trials testing durations of tamoxifen longer than 5 years have not shown additional benefit, but the ATLAS (Adjuvant Tamoxifen, Longer Against Shorter) trial, reported in 2007, showed a small but significant reduction in risk of recurrence with 10 compared with 5 years of tamoxifen therapy. Aromatase inhibitors (AIs) improve relapse-free survival (RFS) in postmenopausal women when they are used sequentially with, or replace, tamoxifen for a total of 5 years of therapy. Extension of endocrine therapy to 10 years in the National Cancer Institute of Canada Clinical Trials Group MA.17 trial demonstrated that 5 years of letrozole therapy following 5 years of tamoxifen therapy results in an improvement in RFS, but not overall survival, in postmenopausal women. Trials testing durations of AI therapy for longer than 5 years are ongoing. Selection of candidates for extended endocrine therapy should balance recurrence risk, toxicity of treatment, and comorbidities that might impact life expectancy and risk of side effects. PMID- 19561006 TI - A 2009 update on the treatment of patients with hormone receptor-positive breast cancer. AB - In up to 75% of breast cancers, estrogen receptor (ER) signaling is a key promoter of tumor proliferation, and inhibition of this pathway has clear therapeutic efficacy. The principal clinical means of inhibiting ER signaling comprise selective ER modulators, such as tamoxifen, that act as partial receptor agonists; measures to reduce the circulating level of estrogen, including ovarian ablation, gonadotropin-releasing hormone analogues, and aromatase inhibition; and antagonism and downregulation of ER by the antiestrogen fulvestrant. Each of these therapies is effective in a proportion of ER-positive breast cancers, but de novo and acquired resistance remain significant problems. Emerging knowledge of the biology of ER signaling will provide insights into the mechanisms of resistance and help guide development of therapeutic strategies to maximize response. This review summarizes the contemporary treatment of early-stage and advanced ER-positive breast cancer in premenopausal and postmenopausal women, with an emphasis on recently published or presented data. Mechanisms of resistance to endocrine interventions and trials exploring strategies to overcome them will also be discussed. PMID- 19561007 TI - An effective method for hemoglobin E detection: DEAE sepharose microcolumn. AB - Hemoglobin E (Hb E) is a variant hemoglobin that can lead to considerable morbidity when it occurs in a compound heterozygous state with beta-thalassemia. Therefore, its detection is important because it permits antenatal counseling. Hb E is prevalent in economically weaker regions of the world. Thus, its recognition is often hindered by the high costs of sophisticated Hb E-detection assays. We developed a simple visual test based on identifying Hb E in blood samples with DEAE Sepharose microcolumn chromatography. The diagnostic accuracy of our new Hb E microcolumn assay was 100% for sensitivity, specificity, and positive and negative predictive values. Furthermore, the Hb E microcolumn assay is rapid, reproducible, and economical; hence, it offers affordable screening for Hb E in less well-equipped laboratories. PMID- 19561008 TI - Usefulness of immature platelet fraction for the clinical evaluation of myelodysplastic syndromes. AB - Ratios of young platelets or reticulated platelets can be routinely obtained as an immature platelet fraction (IPF) with the XE-2100 automated hematology analyzer (Sysmex, Kobe, Japan). We combined IPF analysis of 31 patients with myelodysplastic syndrome (MDS) with a complete blood count, a bone marrow examination, and a chromosome analysis. The patients with >40 x 10(9)/L platelets were classified as group A, and those with > or =40 x 10(9)/L were placed in group B. The 2 groups were subclassified as A1 or B1 for patients with an IPF of <10% and as A2 or B2 for those with an IPF > or =10%. Categories A1, A2, B1, and B2 comprised 12 patients, 6 patients, 7 patients, and 6 patients, respectively. Patients with a relatively high IPF (>10%) (category A2 or B2) showed distinctive characteristics. Group B2 showed a higher frequency of chromosomal abnormalities than B1 (P = .029), and group A2 tended to show a higher incidence of clinical improvement than A1 (P = .08). IPF determination may be clinically useful for the assessment of prognosis for MDS patients. PMID- 19561010 TI - Medical adherence in young adolescents with spina bifida: longitudinal associations with family functioning. AB - OBJECTIVE: The purpose of this study was 2-fold: (1) to explore the transfer of responsibility of medical tasks from parent to child during the transition to adolescence, and (2) to examine the associations between family functioning and medical adherence in youth with spina bifida. METHODS: Seventy families of children with spina bifida participated in this study. Data were collected during family interaction sessions by using questionnaires completed by mothers, fathers, youth, teachers and health professionals. RESULTS: Findings suggest that responsibility for medical regimens transfers gradually from parent to child over time. Additionally, family conflict and cohesion were correlated with medical adherence. Finally, family conflict over medical issues was related to a decrease in medical adherence over time. CONCLUSIONS: Results suggest that as youth take more responsibility over their medical regimens, family conflict regarding medical issues becomes a contributor to their adherence behaviors. Interventions that target family conflict may facilitate adherence behaviors. PMID- 19561011 TI - Functional and genetic diversity of mycorrhizal fungi from single plants of Caladenia formosa (Orchidaceae). AB - BACKGROUND AND AIMS: Mycorrhizal associations are essential to the plant kingdom. The largest flowering plant family, the Orchidaceae, relies on mycorrhizal fungi for germination, growth and survival. Evidence suggests varying degrees of fungal host specificity based on a single fungal isolate from a single plant. This paper shows for the first time the diversity of endophytes colonizing in a single plant over consecutive years and the functional significance of this diversity. METHODS: Stem-collars of Caladenia formosa were collected in different seasons and years. Mycorrhizal fungi isolated were tested for their efficacy to induce leafing and genetically determined using ITS-RFLP and sequencing. RESULTS: Multiple mycorrhizal fungi were repeatedly isolated from a single collar that displayed varying effectiveness in germination percentages and adult leaf length. Additional factors contributed to the isolation of effective mycorrhizal fungi; fungal collection season, year of collection and individual isolates. Surface sterilization only improved the number of isolated mycorrhizal fungi. Dual inoculation did not increase germination. All 59 mycorrhizal fungi effective in germinating seed belonged to one clearly defined ITS (internal transcribed spacer) clade and clustered close to Sebacina vermifera (79-89 % homology). Isolates resulting in the greatest germination were not necessarily those resulting in the greatest survival and growth 1 year after germination. CONCLUSION: Single orchid plants contained multiple mycorrhizal fungal strains of one species that had diverse functional differences. These results suggest that our current knowledge of fungal-host specificity may be incomplete due to experimental and analytical limitations. It also suggests that the long-term effectiveness of a mycorrhizal fungus or fungi could only be found by germination and longer-term growth tests rather than genetically. PMID- 19561012 TI - Mental health policy in South Africa: development process and content. AB - INTRODUCTION: Mental health is increasingly acknowledged as a crucial public health issue in South Africa (SA). However, it is not given the priority it deserves on policy agendas in this and many other low- and middle-income countries. The aim of this analysis is to describe the content of mental health policy and the process of its development in SA. METHODS: Quantitative data regarding SA's mental health system were gathered using the World Health Organization (WHO) Assessment Instrument for Mental Health Systems. The WHO Checklist for Mental Health Policy and Plans was completed for SA's 1997 mental health policy guidelines. Semi-structured interviews provided understanding of processes, underlying issues and interactions between key stakeholders in mental health policy development. RESULTS: There is uncertainty at provincial level regarding whether the 1997 policy guidelines should be considered national policy. At national level the guidelines are not recognized as policy, and a new policy is currently being developed. Although the guidelines were developed through wide consultation and had approval through national policy development processes, difficulties were encountered with dissemination and implementation at provincial level. The principles of these policy guidelines conform to international recommendations for mental health care and services but lack clear objectives. DISCUSSION: The process of mental health policy implementation has been hindered by the low priority given to mental health, varying levels of seniority of provincial mental health coordinators, limited staff for policy and planning, varying technical capacity at provincial and national levels, and reluctance by some provincial authorities to accept responsibility for driving implementation. CONCLUSION: These findings highlight the importance of national leadership in the development of new mental health policy, communication between national and provincial levels, the need for provincial structures to take responsibility for implementation, and capacity building to enable policy makers and planners to develop, monitor and implement policy. PMID- 19561013 TI - Cardiopulmonary arrest in pregnancy: two case reports of successful outcomes in association with perimortem Caesarean delivery. AB - Cardiac arrest in pregnancy is a rare event in which the speed of the response and attention to a number of pregnancy-specific interventions is crucial to the outcome. The commencement of a perimortem Caesarean delivery within 4 min of the onset of the arrest has been recommended as a technique to potentially improve survival in both the mother and the fetus but presents significant logistical challenges to the health-care facility. In this report, we describe two cases of cardiac arrest in pregnancy in which a perimortem Caesarean was performed as part of the resuscitation process and was associated with excellent maternal and neonatal outcomes. We discuss some of the issues surrounding the performance of a perimortem Caesarean delivery that were relevant to this case, including experience from the training that is provided in our institution. PMID- 19561014 TI - Analgesic efficacy of ultrasound-guided transversus abdominis plane block in patients undergoing open appendicectomy. AB - BACKGROUND: Transversus abdominis plane (TAP) block is a new regional anaesthetic technique that blocks abdominal neural afferents by introducing local anaesthetic into the neuro-fascial plane between the internal oblique and the transversus abdominis muscles. We evaluated its analgesic efficacy in patients undergoing open appendicectomy in a randomized controlled double-blinded clinical trial. METHODS: Fifty-two adult patients undergoing open appendicectomy were randomized to undergo standard care (n=26) or to undergo a right-sided TAP block with bupivacaine (n=26). In addition, all patients received patient-controlled i.v. morphine analgesia, regular acetaminophen, and non-steroidal anti-inflammatory drug, as required, in the postoperative period. All patients received standard anaesthetic, and after induction of anaesthesia, the TAP group received an ultrasound-guided unilateral TAP block. Each patient was assessed after operation by a blinded investigator at 30 min and 24 h after surgery. RESULTS: Ultrasound guided TAP block significantly reduced postoperative morphine consumption in the first 24 h [mean (sd) 28 (18) vs 50 (19) mg, P<0.002]. Postoperative visual analogue scale pain scores were also reduced in the TAP block group soon after surgery [median (IQR) 4.5 (3-5.3) vs 8.5 (7.5-10), P<0.001] and at 24 h [5.2 (4 6.2) vs 8 (7-8.5), P<0.001]. There were no complications attributable to the TAP block. CONCLUSIONS: Ultrasound-guided TAP block holds considerable promise as a part of a balanced postoperative analgesic regimen for patients undergoing open appendicectomy. PMID- 19561015 TI - The impact of incomplete knowledge on evaluation: an experimental benchmark for protein function prediction. AB - MOTIVATION: Rapidly expanding repositories of highly informative genomic data have generated increasing interest in methods for protein function prediction and inference of biological networks. The successful application of supervised machine learning to these tasks requires a gold standard for protein function: a trusted set of correct examples, which can be used to assess performance through cross-validation or other statistical approaches. Since gene annotation is incomplete for even the best studied model organisms, the biological reliability of such evaluations may be called into question. RESULTS: We address this concern by constructing and analyzing an experimentally based gold standard through comprehensive validation of protein function predictions for mitochondrion biogenesis in Saccharomyces cerevisiae. Specifically, we determine that (i) current machine learning approaches are able to generalize and predict novel biology from an incomplete gold standard and (ii) incomplete functional annotations adversely affect the evaluation of machine learning performance. While computational approaches performed better than predicted in the face of incomplete data, relative comparison of competing approaches-even those employing the same training data-is problematic with a sparse gold standard. Incomplete knowledge causes individual methods' performances to be differentially underestimated, resulting in misleading performance evaluations. We provide a benchmark gold standard for yeast mitochondria to complement current databases and an analysis of our experimental results in the hopes of mitigating these effects in future comparative evaluations. AVAILABILITY: The mitochondrial benchmark gold standard, as well as experimental results and additional data, is available at http://function.princeton.edu/mitochondria. PMID- 19561016 TI - Transcriptional landscape estimation from tiling array data using a model of signal shift and drift. AB - MOTIVATION: High-density oligonucleotide tiling array technology holds the promise of a better description of the complexity and the dynamics of transcriptional landscapes. In organisms such as bacteria and yeasts, transcription can be measured on a genome-wide scale with a resolution >25 bp. The statistical models currently used to handle these data remain however very simple, the most popular being the piecewise constant Gaussian model with a fixed number of breakpoints. RESULTS: This article describes a new methodology based on a hidden Markov model that embeds the segmentation of a continuous-valued signal in a probabilistic setting. For a computationally affordable cost, this framework (i) alleviates the difficulty of choosing a fixed number of breakpoints, and (ii) permits retrieving more information than a unique segmentation by giving access to the whole probability distribution of the transcription profile. Importantly, the model is also enriched and accounts for subtle effects such as signal 'drift' and covariates. Relevance of this framework is demonstrated on a Bacillus subtilis dataset. AVAILABILITY: A software is distributed under the GPL. PMID- 19561017 TI - libAnnotationSBML: a library for exploiting SBML annotations. AB - SUMMARY: The Systems Biology Markup Language (SBML) is an established community XML format for the markup of biochemical models. With the introduction of SBML level 2 version 3, specific model entities, such as species or reactions, can now be annotated using ontological terms. These annotations, which are encoded using the resource description framework (RDF), provide the facility to specify definite terms to individual components, allowing software to unambiguously identify such components and thus link the models to existing data resources. libSBML is an application programming interface library for the manipulation of SBML files. While libSBML provides the facilities for reading and writing such annotations from and to models, it is beyond the scope of libSBML to provide interpretation of these terms. The libAnnotationSBML library introduced here acts as a layer on top of libSBML linking SBML annotations to the web services that describe these ontological terms. Two applications that use this library are described: SbmlSynonymExtractor finds name synonyms of SBML model entities and SbmlReactionBalancer checks SBML files to determine whether specifed reactions are elementally balanced. PMID- 19561018 TI - Pindel: a pattern growth approach to detect break points of large deletions and medium sized insertions from paired-end short reads. AB - MOTIVATION: There is a strong demand in the genomic community to develop effective algorithms to reliably identify genomic variants. Indel detection using next-gen data is difficult and identification of long structural variations is extremely challenging. RESULTS: We present Pindel, a pattern growth approach, to detect breakpoints of large deletions and medium-sized insertions from paired-end short reads. We use both simulated reads and real data to demonstrate the efficiency of the computer program and accuracy of the results. AVAILABILITY: The binary code and a short user manual can be freely downloaded from http://www.ebi.ac.uk/ approximately kye/pindel/. CONTACT: k.ye@lumc.nl; zn1@sanger.ac.uk. PMID- 19561020 TI - Error control variability in pathway-based microarray analysis. AB - MOTIVATION: The decision to commit some or many false positives in practice rests with the investigator. Unfortunately, not all error control procedures perform the same. Our problem is to choose an error control procedure to determine a P value threshold for identifying differentially expressed pathways in high throughput gene expression studies. Pathway analysis involves fewer tests than differential gene expression analysis, on the order of a few hundred. We discuss and compare methods for error control for pathway analysis with gene expression data. RESULTS: In consideration of the variability in test results, we find that the widely used Benjamini and Hochberg's (BH) false discovery rate (FDR) analysis is less robust than alternative procedures. BH's error control requires a large number of hypothesis tests, a reasonable assumption for differential gene expression analysis, though not the case with pathway-based analysis. Therefore, we advocate through a series of simulations and applications to real gene expression data that researchers control the number of false positives rather than the FDR. PMID- 19561019 TI - A pipeline for the quantitative analysis of CG dinucleotide methylation using mass spectrometry. AB - MOTIVATION: DNA cytosine methylation is an important epigenetic regulator, critical for mammalian development and the control of gene expression. Numerous techniques using either restriction enzyme or affinity-based approaches have been developed to interrogate cytosine methylation status genome-wide, however these assays must be validated by a more quantitative approach, such as MALDI-TOF mass spectrometry of bisulphite-converted DNA (commercialized as Sequenom's EpiTYPER assay using the MassArray system). Here, we present an R package ('MassArray') that assists in assay design and uses the standard Sequenom output file as the input to a pipeline of analyses not available as part of the commercial software. The tools in this package include bisulphite conversion efficiency calculation, sequence polymorphism flagging and visualization tools that combine multiple experimental replicates and create tracks for genome browser viewing. PMID- 19561021 TI - A genetic programming approach for Burkholderia pseudomallei diagnostic pattern discovery. AB - MOTIVATION: Finding diagnostic patterns for fighting diseases like Burkholderia pseudomallei using biomarkers involves two key issues. First, exhausting all subsets of testable biomarkers (antigens in this context) to find a best one is computationally infeasible. Therefore, a proper optimization approach like evolutionary computation should be investigated. Second, a properly selected function of the antigens as the diagnostic pattern which is commonly unknown is a key to the diagnostic accuracy and the diagnostic effectiveness in clinical use. RESULTS: A conversion function is proposed to convert serum tests of antigens on patients to binary values based on which Boolean functions as the diagnostic patterns are developed. A genetic programming approach is designed for optimizing the diagnostic patterns in terms of their accuracy and effectiveness. During optimization, it is aimed to maximize the coverage (the rate of positive response to antigens) in the infected patients and minimize the coverage in the non infected patients while maintaining the fewest number of testable antigens used in the Boolean functions as possible. The final coverage in the infected patients is 96.55% using 17 of 215 (7.4%) antigens with zero coverage in the non-infected patients. Among these 17 antigens, BPSL2697 is the most frequently selected one for the diagnosis of Burkholderia Pseudomallei. The approach has been evaluated using both the cross-validation and the Jack-knife simulation methods with the prediction accuracy as 93% and 92%, respectively. A novel approach is also proposed in this study to evaluate a model with binary data using ROC analysis. PMID- 19561022 TI - Comparative study on ChIP-seq data: normalization and binding pattern characterization. AB - MOTIVATION: Antibody-based Chromatin Immunoprecipitation assay followed by high throughput sequencing technology (ChIP-seq) is a relatively new method to study the binding patterns of specific protein molecules over the entire genome. ChIP seq technology allows scientist to get more comprehensive results in shorter time. Here, we present a non-linear normalization algorithm and a mixture modeling method for comparing ChIP-seq data from multiple samples and characterizing genes based on their RNA polymerase II (Pol II) binding patterns. RESULTS: We apply a two-step non-linear normalization method based on locally weighted regression (LOESS) approach to compare ChIP-seq data across multiple samples and model the difference using an Exponential-Normal(K) mixture model. Fitted model is used to identify genes associated with differential binding sites based on local false discovery rate (fdr). These genes are then standardized and hierarchically clustered to characterize their Pol II binding patterns. As a case study, we apply the analysis procedure comparing normal breast cancer (MCF7) to tamoxifen-resistant (OHT) cell line. We find enriched regions that are associated with cancer (P < 0.0001). Our findings also imply that there may be a dysregulation of cell cycle and gene expression control pathways in the tamoxifen resistant cells. These results show that the non-linear normalization method can be used to analyze ChIP-seq data across multiple samples. AVAILABILITY: Data are available at http://www.bmi.osu.edu/~khuang/Data/ChIP/RNAPII/. PMID- 19561023 TI - Growth-differentiation factor-15 is an independent marker of cardiovascular dysfunction and disease in the elderly: results from the Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) Study. AB - AIMS: Growth-differentiation factor-15 (GDF-15) is emerging as an independent prognostic biomarker in patients with cardiovascular (CV) disease. Little is known about the pathophysiological basis for the close association of GDF-15 to future CV events. We hypothesized that GDF-15 is related to underlying CV pathologies. METHODS AND RESULTS: To relate the levels of GDF-15 to indices of CV dysfunction and disease in elderly individuals, serum levels of GDF-15 were measured in 1004 subjects aged 70 years from the Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) study. Carotid intima-media thickness and plaque burden, and left ventricular (LV) geometry and function were assessed by ultrasound. Endothelial function was evaluated in forearm resistance vessels and in the brachial artery by venous occlusion plethysmography and ultrasound imaging, respectively. Elevated levels of GDF-15 were related to several CV risk factors (male gender, current smoking, body mass index, waist circumference, diabetes, fasting glucose, triglycerides, and low HDL cholesterol). After adjustment for CV risk factors, increased levels of GDF-15 were associated with reduced endothelium-dependent vasodilation in resistance vessels, plaque burden, LV mass and concentric LV hypertrophy, reduced LV ejection fraction, and clinical manifestations of coronary artery disease and heart failure. CONCLUSION: GDF-15 carries information on CV dysfunction and disease that is not captured by traditional CV risk factors in elderly individuals. PMID- 19561024 TI - Randomized controlled trial on the cardioprotective effect of bone marrow cells in patients undergoing coronary bypass graft surgery. AB - AIMS: This randomized study investigates whether bone marrow cells (BMCs) can reduce ischaemic injury during cardiac surgery. METHODS AND RESULTS: Forty-four elective coronary artery bypass grafting patients were randomized to control group or BMCs group (whereby autologous BMCs were administered with each dose of cardioplegia antegradely into the coronaries). Troponin I and CK-MB were measured during the first 48 h after surgery and were not significantly different between the control and BMCs groups. The role of cardiopulmonary bypass (CPB) on the cardioprotective effects of BMCs was also studied using an in vitro model of stimulated ischaemia and reoxygenation on right atrial appendages obtained from controls either before or 10 min after the initiation of CPB. Bone marrow cells significantly reduced myocardial injury in muscles obtained prior to CPB. This effect was comparable with ischaemic preconditioning (IP), although their combination did not afford additional benefit. However, when muscles were harvested after CPB, myocardial injury in the ischaemic group alone was less, and BMCs or IP did not exert further protection. CONCLUSION: Bone marrow cells did not afford additional benefit when used as an additive to cardioplegia during CPB. However, BMCs offer cardioprotection as potent as IP, when the heart is not subjected to stress, such as CPB, that per se can precondition the myocardium. PMID- 19561025 TI - Magnetic resonance investigations in Brugada syndrome reveal unexpectedly high rate of structural abnormalities. AB - AIMS: Recent data suggest that sub-clinical structural abnormalities may be part of the Brugada syndrome (BrS) phenotype, a disease traditionally thought to occur in the structurally normal heart. In this study, we carried out detailed assessment of cardiac morphology and function using cardiac magnetic resonance imaging (CMRI). METHODS AND RESULTS: Thirty consecutive patients with BrS were compared with 30 sex- (26/4 male/female), body surface area- (+/-0.2 m(2)), and age-matched (+/-5 years) normal volunteers. CMRI exam included long- and short axis ECG-gated breath-hold morphological T1-TSE sequences for fatty infiltration and cine-SSFP sequences for kinetic assessment. Fatty infiltration was not found in any subject. Patients with BrS compared with normal subjects showed higher incidence of mild right ventricle (RV) wall-motion abnormalities [15 (50%) vs. 5 (17%) subjects (P = 0.006) with reduced radial fractional shortening in more than two segments], reduction of outflow tract ejection fraction (49 +/- 11% vs. 55 +/ 10%; P = 0.032), enlargement of the inflow tract diameter (46 +/- 4 vs. 41 +/- 5 mm, P < 0.001 in short-axis; 46 +/- 4 vs. 42 +/- 5 mm, P = 0.001 in four-chamber long-axis view) and area (22 +/- 2 vs. 20 +/- 3 cm(2); P = 0.050), and of global RV end-systolic volume (34 +/- 10 vs. 30 +/- 6 mL/m(2); P = 0.031) but comparable outflow tract dimensions, global RV end-diastolic volume, left ventricle parameters, and atria areas. CONCLUSION: CMRI detects a high prevalence of mild structural changes of the RV, and suggests further pathophysiological complexity in BrS. Prospective studies to assess the long-term evolution of such abnormalities are warranted. PMID- 19561026 TI - Intra-aortic phased-array imaging: new guiding tool for transcatheter aortic valve implantation. PMID- 19561027 TI - Improvement in left ventricular filling properties after relief of right ventricle to pulmonary artery conduit obstruction: contribution of septal motion and interventricular mechanical delay. AB - AIMS: To investigate the impact of relief of right ventricle (RV) to pulmonary artery (PA) conduit obstruction on septal motion and ventricular interaction and its functional implications for left ventricular (LV) filling properties. METHODS AND RESULTS: In 20 consecutive patients with congenital heart disease and RV to PA conduit obstruction, the following were prospectively assessed before and after percutaneous pulmonary valve implantation (PPVI): the septal curvature and LV volumes throughout the cardiac cycle by magnetic resonance imaging; RV to LV mechanical delay by 2D-echocardiographic strain imaging; and objective exercise capacity. Percutaneous pulmonary valve implantation led to a reduction in RV to LV mechanical delay (127.9 +/- 50.9 vs. 37.7 +/- 35.6 ms; P < 0.001) and less LV septal bowing in early LV diastole (septal curvature: -0.11 +/- 0.11 vs. 0.07 +/- 0.13 cm(-1); P < 0.001). Early LV diastolic filling (first one-third of diastole) increased significantly (17.5 +/- 9.4 to 30.4 +/- 9.4 mL/m(2); P < 0.001). The increase in early LV diastolic filling correlated with the reduction in RV to LV mechanical delay (r = -0.68; P = 0.001) and change in septal curvature (r = 0.71; P < 0.001). In addition, the improvement in peak oxygen uptake (56.0 +/- 16.0 vs. 64.1 +/- 13.7% of predicted; P < 0.001) was associated with the increase in early LV diastolic filling (r = 0.69; P = 0.001). CONCLUSION: Relief of RV to PA conduit obstruction significantly improves early LV filling properties. This is attributed to more favourable septal motion and reduction in interventricular mechanical delay. PMID- 19561028 TI - The heat is off: immunosuppression for myocarditis revisited. PMID- 19561029 TI - Peak exercise responses in heart failure: back to basics. PMID- 19561030 TI - Galectin-1 stimulates monocyte chemotaxis via the p44/42 MAP kinase pathway and a pertussis toxin-sensitive pathway. AB - Galectin-1, the prototype of a family of beta-galactoside-binding proteins, has been implicated in a wide variety of biological processes. Data presented herein show that galectin-1 stimulates monocyte migration in a dose-dependent manner but is not chemotactic for macrophages. Galectin-1-induced monocyte chemotaxis is blocked by lactose and inhibited by an anti-galectin-1 antibody but not by nonspecific antibodies. Furthermore, galectin-1-mediated monocyte migration was significantly inhibited by MEK inhibitors in a rapid, time-dependent manner suggesting that MAP kinase pathways are involved in galectin-1. Migration was also almost completely blocked by pertussis toxin implying G-protein involvement in the galectin-1-induced chemotaxis. These results demonstrate a role for galectin-1 in monocyte chemotaxis which differs from galectin-3 in that macrophages are nonresponsive. Furthermore, our observations suggest that galectin-1 may be involved in chemoattraction at sites of inflammation in vivo and may contribute to disease processes such as atherosclerosis. PMID- 19561031 TI - Role of N-glycosylation of the SEA module of rodent Muc3 in posttranslational processing of its carboxy-terminal domain. AB - A prominent feature of the rodent Muc3 SEA module is the precursor cleavage event that segregates the O-glycosylated N-terminal fragment and transmembrane domain into the noncovalently attached heterodimer. There are seven potential N glycosylation sites that occur in a cluster in the SEA module of Muc3. However, it is unknown if these sites are modified or what the function of these N-glycans may be in the SEA module. Our data show that the proteolytic cleavage of the rodent Muc3 SEA module was partially prevented by treatment with tunicamycin, an inhibitor of N-glycosylation. Each single mutant of the seven N-glycosylation sites (N1A, N2A, N3A, N4A, N5A, N6A, and N7A) and multiple mutants, including double (N34A) and triple (N345A) mutants, and mutants with four (N3457A), five (N34567A), six (N134567A and N234567A), seven (N1234567A) mutations, confirmed that all seven of these potential sites are N-glycosylated simultaneously. The proteolytic cleavage of the SEA module was not affected when it lacked only one, two, or three N-glycans, but was partially inhibited when lacking four, five, and six N-glycans. In all, 2%, 48%, 85%, and 73% of the products from N3457A, N34567A, N134567A, and N234567A transfectants, respectively, remained uncleaved. The proteolytic cleavage was completely prevented in the N1234567A transfectant, which eliminated all seven N-glycans in the SEA module. The interaction of the heterodimer was independent of the N-glycans within the rodent Muc3 SEA module. Thus, the N-glycosylation pattern constituted a control point for the modulation of the proteolytic cleavage of the SEA module. PMID- 19561032 TI - Exposure to inhalable dust, endotoxins, beta(1->3)-glucans, and airborne microorganisms in horse stables. AB - OBJECTIVES: Workers in horse stables are likely exposed to high levels of organic dust. Organic dusts play a role in increased risk of inflammatory reactions and are associated with respiratory diseases. The aim of this study was to investigate dust, endotoxin, beta(1-->3)-glucan, and culturable microorganisms exposure levels in horse stables. METHODS: Ambient (n = 38) and personal (n = 42) inhalable dust samples were collected using PAS-6 sampling heads. As a special measurement, we included sampling near the horses' heads. Samples were analyzed for endotoxin and beta(1-->3)-glucan by Limulus amebocyte lysate assay and an inhibition enzyme immunoassay, respectively. Culturable bacteria and fungi were collected with an Anderson impactor. RESULTS: Geometric means (GMs) of personal exposure to dust, endotoxin, and beta(1-->3)-glucan were 1.4 mg m(-3) (range 0.2 9.5), 608 EU m(-3) (20-9846), and 9.5 microg m(-3) (0.4-631 microg m(-3)), respectively. Exposure levels in the morning shift were higher compared to other shifts. The GMs (ranges) of culturable bacteria and fungi were 3.1 x 10(3) colony forming unit (CFU) m(-3) (6.7 x 10 to 1.9 x 10(4)) and 1.9 x 10(3) CFU m(-3) (7.4 x 10 to 2.4 x 10(4)), respectively. Variance components for endotoxin and beta(1- >3)-glucan were considerably higher than for dust. Based on dummy variable in a mixed regression analysis, the predominant task explaining exposure levels of dust, endotoxin, and beta(1-->3)-glucan was sweeping the floor. For beta(1-->3) glucan, feeding the horse was also an important determinant. CONCLUSION: Dust, endotoxin, and beta(1-->3)-glucan exposure are considerable in horse stables. Bacterial and fungal exposure levels were moderate. Endotoxin exposures were above the Dutch proposed standard limits, suggesting workers in horse stables to be at risk of adverse health effects. PMID- 19561033 TI - The prevalence of depression in hereditary spastic paraplegia. AB - OBJECTIVE: To evaluate the prevalence of depression and sensitivity and specificity of the single-item interview 'Are you depressed?' for people with hereditary spastic paraplegia in Estonia. DESIGN: Single-item interview 'Are you depressed?' was used as a screening question for depression; all participants then completed the Beck Depression Inventory. SETTING: People with hereditary spastic paraplegia identified from the epidemiological database who agreed to participate in the study. MAIN MEASURES: Beck Depression Inventory, clinical interview. RESULTS: The epidemiological database consisted of 59 patients with clinically confirmed diagnosis of hereditary spastic paraplegia. Forty-eight of these consented to participate in the study. The Beck Depression Inventory score was higher than cut-off point in 58% (28/48) and lower in 42% (20/48). Of the study group, 44% (21/48) had mild, 13% (6/48) moderate and one person revealed severe depression. There was a statistically significant correlation between Beck Depression Inventory score and level of mobility; no other significant correlations with other measures were detected. Of the participants, 54% (26/48) had subjective complaints about depression and answered 'Yes' to the single-item interview 'Are you depressed?'. The sensitivity of the one-item interview in the hereditary spastic paraplegia group was 75% and specificity 75%. CONCLUSIONS: Our results show that mild depression is prevalent among people with hereditary spastic paraplegia. Although the single question may be helpful, it cannot be relied upon entirely when assessing a person for depression. PMID- 19561034 TI - The effect of an intensive exercise programme on leg function in chronic stroke patients: a pilot study with one-year follow-up. AB - OBJECTIVE: To investigate the effect of two weeks of intensive exercise on leg function in chronic stroke patients and to evaluate the feasibility of an intensive exercise programme in a group setting. DESIGN: Pilot study with one group pre-test post-test design with two pre-tests and one-year follow-up. SETTING: Inpatient rehabilitation hospital. SUBJECTS: Twelve hemiparetic patients completed the intervention. Ten patients participated at one-year follow-up. INTERVENTION: Six hours of daily intensive exercise for two weeks with focus on weight-shifting towards the affected side and increased use of the affected extremity during functional activities. An insole with nubs in the shoe of the non-paretic limb was used to reinforce weight-shift toward the affected side. MAIN MEASURES: Timed Up and Go, Four Square Step Test, gait velocity, gait symmetry and muscle strength in knee and ankle muscles. RESULTS: Maximal gait velocity (P = 0.002) and performance time (seconds) on Timed Up and Go (mean, SD; 12.2, 3.8 vs. 9.4, 3.2) and Four Square Step Test improved from pre- to post-test (P = 0.005). Improvements remained significant at follow-up. Preferred gait velocity and gait symmetry remained unchanged. Knee extensor (P<50.009) and flexor (P<50.001) strength increased bilaterally from pre- to post-test but only knee flexor strength remained significant at follow-up. Ankle dorsi flexor (P = 0.02) and plantar flexor (P<0.001) strength increased on paretic side only (not tested at follow-up). CONCLUSION: Intensive exercise for lower extremity is feasible in a group setting and was effective in improving ambulatory function, maximal gait velocity and muscle strength in chronic stroke patients. Most improvements persisted at the one-year follow-up. PMID- 19561035 TI - Vibration therapy in multiple sclerosis: a pilot study exploring its effects on tone, muscle force, sensation and functional performance. AB - OBJECTIVE: To examine the effectiveness of whole body vibration (WBV) on tone, muscle force, sensation and functional performance in people with multiple sclerosis. DESIGN: A randomized cross-over pilot study. SETTING: Revive MS Support Therapy Centre. Glasgow, UK. SUBJECTS: Sixteen people with multiple sclerosis were randomly allocated to one of two groups. INTERVENTION: Group 1 received four weeks of whole body vibration plus exercise three times per week, two weeks of no intervention and then four weeks of exercise alone three times per week. Group 2 were given the two treatment interventions in the reverse order to group 1. MAIN MEASURES: Ten-metre walk, Timed Up and Go Test, Modified Ashworth Scale, Multiple Sclerosis Spasticity Scale (MSSS-88), lower limb muscle force, Nottingham Sensory Assessment and Multiple Sclerosis Impact Scale (MSIS 29) were used before and after intervention. RESULTS: The exercise programme had positive effects on muscle force and well-being, but there was insufficient evidence that the addition of whole body vibration provided any further benefit. The Modified Ashworth Scale was generally unaffected by either intervention, although, for each group, results from the MSSS-88 showed whole body vibration and exercises reduced muscle spasms (P = 0.02). Although results for the 10-m walk and Timed Up and Go Test improved, this did not reach statistical significance (P = 0.56; P = 0.70, respectively). For most subjects sensation was unaffected by whole body vibration. CONCLUSION: Exercise may be beneficial to those with multiple sclerosis, but there is limited evidence that the addition of whole body vibration provides any additional improvements. Further larger scale studies into the effects of whole body vibration in people with multiple sclerosis are essential. PMID- 19561036 TI - Retrospective and prospective studies of hepatitis B virus reactivation in malignant lymphoma with occult HBV carrier. AB - BACKGROUND: In surface antigen of hepatitis B virus (HBsAg)-positive carrier for anticancer treatment of malignant lymphoma, it is well recognized that reactivation of hepatitis B virus (HBV) occasionally occurs. However, there have been only a few studies of HBV reactivation in serum HBsAg-negative and hepatitis B core antigen (HBcAb)-positive occult HBV carriers. We looked at both retrospective and prospective studies to determine the prevalence, clinical course and risk factor of HBV reactivation during chemotherapy in lymphoma patients. PATIENTS AND METHODS: Forty-eight of 127 (37.8%) lymphoma patients were HBsAg negative and HBcAb positive, and 24 of these patients were then given liver function tests and HBsAg tests monthly and serum HBV DNA every 3 months. RESULTS: HBV reactivation was observed in two patients (4.1%) who had received intensive chemotherapy including steroid and rituximab. Immediate administration of entecavir therapy after elevation of HBV DNA level was conducted, and this resulted in reduction of it and improvement of liver function test. CONCLUSIONS: Rituximab plus steroid-containing regimens may increase the risk of HBV reactivation in HBsAg-negative and HBcAb-positive lymphoma patients. More ambitious prospective studies are required to establish clinically useful or cost effective follow-up methods for control of HBV reactivation in lymphoma patients with occult HBV infection. PMID- 19561037 TI - A phase II study of palonosetron combined with dexamethasone to prevent nausea and vomiting induced by highly emetogenic chemotherapy. AB - BACKGROUND: This is a randomized, double-blind, dose-ranging study in patients receiving highly emetogenic chemotherapy (HEC) to evaluate the safety, efficacy, and pharmacokinetics of palonosetron, in combination with dexamethasone. MATERIALS AND METHODS: We randomized 233 patients to receive palonosetron as a single i.v. bolus dose of 0.075, 0.25, or 0.75 mg before administration of HEC. Dexamethasone (12-16 mg i.v. on day 1, 8 mg i.v. on day 2, and 4-8 mg i.v. on day 3) was administered for prophylactic antiemesis. Pharmacokinetics of palonosetron was analyzed in 24 patients. RESULTS: In this study, all patients were given > or =50 mg/m(2) cisplatin, which was considered to be HEC. No significant differences in complete response (CR: no emesis and no rescue medication) rates were found in the first 24 h between the 0.075-, 0.25-, and 0.75-mg groups (77.6%, 81.8%, and 79.5%, respectively). In the 120-h period of overall observation, CR rates increased in a dose-dependent manner. In the 0.75-mg group, we observed a significantly longer time to treatment failure than in the 0.075-mg group (median time >120 versus 82.0 h, P = 0.038). Palonosetron was tolerated well and did not show any dose-related increase in adverse effects. CONCLUSIONS: Palonosetron at doses of 0.25 and 0.75 mg was shown to be effective in preventing chemotherapy induced nausea and vomiting with high CR rates of patients treated with HEC in Japan. All tested doses of palonosetron were tolerated well. PMID- 19561039 TI - Targeting of cardiac autonomic plexus for modulation of intracardiac neural tone. AB - AIMS: Ventricular rate control is considered as an initial choice of therapy in many patients with atrial fibrillation (AF). We could previously show that electrostimulation of the right inferior ganglionated plexus (RIGP), which supplies the AV node, instantly decreases ventricular rate during AF. This study describes the development of a technique to reliably implant a chronic lead inside the RIGP. METHODS AND RESULTS: In nine mongrel dogs with AF, the RIGP was identified by neuromapping with probatory high-frequency stimulation (20 Hz) over steerable electrode catheters until a significant ventricular rate slowing was achieved. Then an active fixation, permanent pacemaker lead was fixed closed to the mapping catheter left in place as anatomical marker. Initially (n = 4) available guiding catheters and steerable lead stylets were employed to navigate and anchor the lead, which resulted in repetitive screw-in attempts. Therefore, a guiding catheter was developed, which allowed angiography, lead advancement through its lumen, and probatory neurostimulation over its tip. This tool allowed lead delivery within 40 min (n = 5). Neurostimulation via the permanent lead elicited negative dromotropic effects with stimulation frequency, voltage, and impulse duration as determinants of stimulation efficacy. CONCLUSION: Active fixation of a permanent pacing lead inside the RIGP is feasible without thoracotomy. Thereby, ventricular rate control during AF can be achieved with stimulus voltages applied for myocardial electrostimulation. PMID- 19561038 TI - Cancer and thrombosis: implications of published guidelines for clinical practice. AB - Cancer is a frequent finding in patients with thrombosis, and thrombosis is much more prevalent in patients with cancer, with important clinical consequences. Thrombosis is the second most common cause of death in cancer patients. Venous thromboembolism (VTE) in cancer is also associated with a high rate of recurrence, bleeding, a requirement for long-term anticoagulation, and worsened quality of life. Risk factors for cancer-associated VTE include particular cancer types, chemotherapy (with or without antiangiogenic agents), the use of erythropoietin-stimulating agents, the presence of central venous catheters, and surgery. Novel risk factors include platelet and leukocyte counts and tissue factor. A risk model for identifying cancer patients at highest risk for VTE has recently been developed. Anticoagulant therapy is safe and efficacious for prophylaxis and treatment of VTE in patients with cancer. Available anticoagulants include warfarin, heparin, and low-molecular weight heparins (LMWHs). LMWHs represent the preferred therapeutic option for VTE prophylaxis and treatment. Their use may be associated with improved survival in cancer, although this issue requires further study. Despite the significant burden imposed by VTE and the availability of effective anticoagulant therapies, many oncology patients do not receive appropriate VTE prophylaxis as recommended by practice guidelines. Improved adherence to guidelines could substantially reduce morbidity, decrease resource use, enhance quality of life, and improve survival in these patients. PMID- 19561040 TI - Cause of recurrent syncope revealed. AB - Implantable loop recorders (ILRs) allow long-time rhythm monitoring in order to correlate unexplained symptoms with possible arrhythmias. In this case report, we describe a patient with unexplained syncope in which the ILR revealed the cause. PMID- 19561041 TI - Risk of contamination of germplasm during cryopreservation and cryobanking in IVF units. AB - Cryopreservation of sperm, embryos and, more recently, oocytes plays an important and increasing role in assisted reproduction, due to improvements of old, and introduction of new technologies. In parallel, concerns are increasing about the technical and biological safety of these procedures. However, published data regarding the confirmed or theoretical hazards of these procedures are sparse and sometimes contradictory. The purpose of this review will summarize data and opinions about one of the most disputed risks, the potential hazard of contamination and disease transmission through cryopreservation. Special attention is concentrated on the weak points of the technology including open vitrification systems, sterilization of liquid nitrogen and safety of commonly used storage tanks including straws and cryovials. Suggestions are also made for practical measures to avoid these dangers while preserving the benefits and perspectives of new cryopreservation technologies. PMID- 19561042 TI - Under-reporting of tobacco use among Bangladeshi women in England. AB - BACKGROUND: This study investigates the prevalence of under-reported use of tobacco among Bangladeshi women and the characteristics of this group. METHODS: The 1999 and 2004 Health Survey for England included 996 Bangladeshi women aged 16 years and above, 302 with a valid saliva sample and 694 without. The main outcome measure was the prevalence of under-reported tobacco use. RESULTS: Fifteen per cent of Bangladeshi women with a saliva sample under-reported their personal tobacco use. Under-reporters were very similar to self-reported users except for being much more likely to report chewing paan without tobacco (47% versus 9%, P < 0.001). Under-reporters differed significantly from cotinine validated non-users in most respects. Regression analyses confirmed that under reporters and self-reported users were similar in age, education level and exposure to passive smoking. Under-reporters were older and less educated than cotinine-validated non-users. Both self-reported users [odds ratio (OR): 0.11, 95% confidence interval (CI): 0.04-0.30] and cotinine-validated non-users (OR: 0.42, 95% CI: 0.20-0.89) were far less likely to report chewing paan without tobacco compared with under-reporters. CONCLUSIONS: Contrary to our a priori hypothesis, under-reporters were not young, British-born, English-speaking women likely to be concealing smoking but resembled self-reported tobacco users except for being much more likely to report chewing paan without tobacco. PMID- 19561043 TI - Restricted thyroglobulin antibody epitope specificities in subjects with type 1 diabetes mellitus. AB - OBJECTIVES: Following iodisation in Sri Lanka we observed a high prevalence of thyroglobulin antibodies (TgAbs) in type 1 diabetic (T1DM) patients. The clinical significance of these TgAbs is uncertain. We sought to obtain a detailed epitope analysis of TgAbs in T1DM patients recruited from diabetes clinics and to compare these with TgAb epitope specificities in patients with autoimmune thyroid disease (AITD) and healthy individuals in that country. DESIGN AND METHODS: We used a panel of 10 Tg-MAbs in competitive ELISA reactions in a prospective study of subjects recruited from Colombo, to determine the epitopes recognised by TgAb positive patients with T1DM (n=58, 34F:24M, median age 16 years), AITD patients (n=42, 33F:9M, median age 37 years) and healthy subjects (n=50, 39F:11M, median age 27 years). The outcomes were a comparison of reactivity with six Tg clusters (I-VI) in these subjects, and the relation of epitope specificity patterns with free thyroxine and TSH. RESULTS: Patients with T1DM and AITD but not healthy control subjects preferentially recognised the immunodominant clusters, I, III and IV. Patients with these narrow epitope specificities had higher median TSH levels (1.60 vs 1.06; P=0.01), and were more frequently positive for antibodies to thyroid peroxidase than those with broad specificities (52.3 vs 7.1%; P=0.004). CONCLUSIONS: The TgAb epitope specificities in euthyroid Sri Lankans with T1DM are similar to AITD patients. TgAb epitope studies may potentially identify T1DM patients at risk of thyroid dysfunction. PMID- 19561044 TI - Influence of prematurity and growth restriction on the adipokine profile, IGF1, and ghrelin levels in cord blood: relationship with glucose metabolism. AB - OBJECTIVE: To determine the influence of gestational age and fetal growth restriction on the cord blood adipokine profile, IGF1, and ghrelin levels, and their relationship with glucose metabolism. STUDY DESIGN: One hundred and ninety newborns (99 preterm and 91 full term) were studied and, according to their anthropometry at birth, classified as small (SGA) or adequate for gestational age (AGA). METHODS: Venous cord blood serum levels of IGF1, IGF binding protein 3 (IGFBP-3), adiponectin, resistin, leptin, soluble leptin receptor (sOB-R), tumoral necrosis factor-alpha, interleukin 6 (IL-6), total ghrelin, and acylated ghrelin were determined and compared between preterm and full-term, as well as between SGA and AGA, newborns. Correlations with newborn weight, gestational age, and homeostatic model assessment (HOMA) index, as an index of insulin resistance, were determined. RESULTS: Preterm newborns had higher HOMA, sOB-R, resistin, and IL-6 and lower IGF1, IGFBP-3, leptin, and adiponectin levels than full-term newborns. SGA had lower IGF1, IGFBP-3, leptin, IL-6, and adiponectin and higher sOB-R and total ghrelin than AGA newborns. Adiponectin and HOMA showed independent positive and negative correlations with gestational age respectively, but not with neonatal weight. Birth weight was correlated positively with IGF1 and leptin levels and negatively with total ghrelin ones. CONCLUSIONS: Our findings suggest that the lack of proper acquisition of adipose tissue by the fetus either due to prematurity or to fetal growth restriction is associated with changes in the cord blood adipokine profile that may contribute to the impairment of glucose metabolism. PMID- 19561045 TI - Light chain-deficient mice produce novel multimeric heavy-chain-only IgA by faulty class switching. AB - Recently, we identified that diverse heavy chain (H-chain)-only IgG is spontaneously produced in light chain (L-chain)-deficient mice (L(-/-) with silenced kappa and lambda loci) despite a block in B cell development. In murine H-chain IgG, the first Cgamma exon, C(H)1, is removed after DNA rearrangement and secreted polypeptides are comparable with camelid-type H-chain IgG. Here we show that L(-/-) mice generate a novel class of H-chain Ig with covalently linked alpha chains, not identified in any other healthy mammal. Surprisingly, diverse H chain-only IgA can be released from B cells at levels similar to conventional IgA and is found in serum and sometimes in milk and saliva. Surface IgA without L chain is expressed in B220(+) spleen cells, which exhibited a novel B cell receptor, suggesting that associated conventional differentiation events occur. To facilitate the cellular transport and release of H-chain-only IgA, chaperoning via BiP association seems to be prevented as only alpha chains lacking C(H)1 are released from the cell. This appears to be accomplished by imprecise class-switch recombination (CSR) from Smu into the alpha constant region, which removes all or part of the Calpha1 exon at the genomic level. PMID- 19561046 TI - Bacillus subtilis-specific poly-gamma-glutamic acid regulates development pathways of naive CD4(+) T cells through antigen-presenting cell-dependent and independent mechanisms. AB - Peripheral naive CD4(+) T cells selectively differentiate to type 1 T(h), type 2 T(h) and IL-17-producing T(h) (T(h)17) cells, depending on the priming conditions. Since these subsets develop antagonistically to each other to elicit subset-specific adaptive immune responses, balance between these subsets can regulate the susceptibility to diverse immune diseases. The present study was undertaken to determine whether poly-gamma-glutamic acid (gamma-PGA), an edible and safe exopolymer that is generated by microorganisms such as Bacillus subtilis, could modulate the development pathways of T(h) subsets. The presence of gamma-PGA during priming promoted the development of T(h)1 and T(h)17 cells but inhibited development of T(h)2 cells. gamma-PGA up-regulated the expression of T-bet and ROR-gammat, the master genes of T(h)1 and T(h)17 cells, respectively, whereas down-regulating the level of GATA-3, the master gene of T(h)2 cells. gamma-PGA induced the expression of IL-12p40, CD80 and CD86 in dendritic cells (DC) and macrophages in a Toll-like receptor-4-dependent manner, and the effect of gamma-PGA on T(h)1/T(h)2 development was dependent on the presence of antigen-presenting cells (APC). Furthermore, gamma-PGA-stimulated DC favored the polarization of naive CD4(+) T cells toward T(h)1 cells rather than T(h)2 cells. In contrast, gamma-PGA affected T(h)17 cell development, regardless of the presence or absence of APC. Thus, these data demonstrate that gamma-PGA has the potential to regulate the development pathways of naive CD4(+) T cells through APC-dependent and -independent mechanisms and to be applicable to treating T(h)2-dominated diseases. PMID- 19561047 TI - Exploring the importance of within-canopy spatial temperature variation on transpiration predictions. AB - Models seldom consider the effect of leaf-level biochemical acclimation to temperature when scaling forest water use. Therefore, the dependence of transpiration on temperature acclimation was investigated at the within-crown scale in climatically contrasting genotypes of Acer rubrum L., cv. October Glory (OG) and Summer Red (SR). The effects of temperature acclimation on intracanopy gradients in transpiration over a range of realistic forest growth temperatures were also assessed by simulation. Physiological parameters were applied, with or without adjustment for temperature acclimation, to account for transpiration responses to growth temperature. Both types of parameterization were scaled up to stand transpiration (expressed per unit leaf area) with an individual tree model (MAESTRA) to assess how transpiration might be affected by spatial and temporal distributions of foliage properties. The MAESTRA model performed well, but its reproducibility was dependent on physiological parameters acclimated to daytime temperature. Concordance correlation coefficients between measured and predicted transpiration were higher (0.95 and 0.98 versus 0.87 and 0.96) when model parameters reflected acclimated growth temperature. In response to temperature increases, the southern genotype (SR) transpiration responded more than the northern (OG). Conditions of elevated long-term temperature acclimation further separate their transpiration differences. Results demonstrate the importance of accounting for leaf-level physiological adjustments that are sensitive to microclimate changes and the use of provenance-, ecotype-, and/or genotype specific parameter sets, two components likely to improve the accuracy of site level and ecosystem-level estimates of transpiration flux. PMID- 19561049 TI - Elevated CO2 significantly delays reproductive development of soybean under Free Air Concentration Enrichment (FACE). AB - The effect of rising atmospheric concentration of carbon dioxide [CO(2)] on the reproductive development of soybean (Glycine max. Merr) has not been evaluated under open-air field conditions. Soybeans grown under Free-Air CO(2) Enrichment (FACE) exhibit warmer canopies due to decreased latent heat loss because of decreased stomatal conductance. According to development models based on accumulated thermal time, or growing degree days ( degrees Cd), increased canopy temperature should accelerate development. The SoyFACE research facility (Champaign, Illinois, USA) was used to test the hypothesis that development is accelerated in soybean when grown in [CO(2)] elevated to 548 micromol mol(-1). Canopy temperature was measured continuously with infrared thermometry, and used in turn to calculate GDD. Opposite to expectation, elevated [CO(2)], while increasing canopy temperature, delayed reproductive development by up to 3 days (P <0.05). Soybean grown in elevated [CO(2)] required approximately 49 degrees Cd more GDD (P <0.05) to complete full bloom stage (R2) and approximately 52 degrees Cd more GDD (P <0.05) to complete the beginning seed (R5) stage, but needed approximately 46 degrees Cd fewer GDD (P <0.05) to complete seed filling (R6). Soybeans grown in elevated [CO(2)] produced significantly more nodes (P <0.01) on the main stem than those grown under current [CO(2)]. This may explain the delay in completion of reproductive development and final maturation of the crop under elevated [CO(2)]. These results show a direct effect of rising [CO(2)] on plant development that will affect both projections of grain supply and may be significant to other species including those in natural communities. PMID- 19561048 TI - Differentially expressed genes between drought-tolerant and drought-sensitive barley genotypes in response to drought stress during the reproductive stage. AB - Drought tolerance is a key trait for increasing and stabilizing barley productivity in dry areas worldwide. Identification of the genes responsible for drought tolerance in barley (Hordeum vulgare L.) will facilitate understanding of the molecular mechanisms of drought tolerance, and also facilitate the genetic improvement of barley through marker-assisted selection or gene transformation. To monitor the changes in gene expression at the transcriptional level in barley leaves during the reproductive stage under drought conditions, the 22K Affymetrix Barley 1 microarray was used to screen two drought-tolerant barley genotypes, Martin and Hordeum spontaneum 41-1 (HS41-1), and one drought-sensitive genotype Moroc9-75. Seventeen genes were expressed exclusively in the two drought-tolerant genotypes under drought stress, and their encoded proteins may play significant roles in enhancing drought tolerance through controlling stomatal closure via carbon metabolism (NADP malic enzyme, NADP-ME, and pyruvate dehydrogenase, PDH), synthesizing the osmoprotectant glycine-betaine (C-4 sterol methyl oxidase, CSMO), generating protectants against reactive-oxygen-species scavenging (aldehyde dehydrogenase,ALDH, ascorbate-dependent oxidoreductase, ADOR), and stabilizing membranes and proteins (heat-shock protein 17.8, HSP17.8, and dehydrin 3, DHN3). Moreover, 17 genes were abundantly expressed in Martin and HS41-1 compared with Moroc9-75 under both drought and control conditions. These genes were possibly constitutively expressed in drought-tolerant genotypes. Among them, seven known annotated genes might enhance drought tolerance through signalling [such as calcium-dependent protein kinase (CDPK) and membrane steroid binding protein (MSBP)], anti-senescence (G2 pea dark accumulated protein, GDA2), and detoxification (glutathione S-transferase, GST) pathways. In addition, 18 genes, including those encoding Delta(l)-pyrroline-5-carboxylate synthetase (P5CS), protein phosphatase 2C-like protein (PP2C), and several chaperones, were differentially expressed in all genotypes under drought; thus they were more likely to be general drought-responsive genes in barley. These results could provide new insights into further understanding of drought-tolerance mechanisms in barley. PMID- 19561050 TI - Over the falls? Rapid evolution of ecotypic differentiation in steelhead/rainbow trout (Oncorhynchus mykiss). AB - Adaptation to novel habitats and phenotypic plasticity can be counteracting forces in evolution, but both are key characteristics of the life history of steelhead/rainbow trout (Oncorhynchus mykiss). Anadromous steelhead reproduce in freshwater river systems and small coastal streams but grow and mature in the ocean. Resident rainbow trout, either sympatric with steelhead or isolated above barrier dams or waterfalls, represent an alternative life-history form that lives entirely in freshwater. We analyzed population genetic data from 1486 anadromous and resident O. mykiss from a small stream in coastal California with multiple barrier waterfalls. Based on data from 18 highly variable microsatellite loci (He = 0.68), we conclude that the resident population above one barrier, Big Creek Falls, is the result of a recent anthropogenic introduction from the anadromous population of O. mykiss below the falls. Furthermore, fish from this above barrier population occasionally descend over the falls and have established a genetically differentiated below-barrier subpopulation at the base of the falls, which appears to remain reproductively isolated from their now-sympatric anadromous ancestors. These results support a hypothesis of rapid evolution of a purely resident life history in the above-barrier population in response to strong selection against downstream movement. PMID- 19561051 TI - Managing hepatitis C virus infection. PMID- 19561052 TI - Government claims measures to cut MRSA and C difficile rates will help reduce other infections. PMID- 19561053 TI - Age for starting cervical cancer screening in England will not be lowered. PMID- 19561054 TI - Wastewater workers and hepatitis A virus infection. AB - BACKGROUND: The main occupational hazard of wastewater workers (WWs) is the direct exposure to the variety of infectious agents present in sewage material, with hepatitis A virus (HAV) being the most frequent one. Most epidemiological studies have shown a higher risk of hepatitis A among WWs, although some studies have produced conflicting evidence. AIMS: To evaluate the hypothesis of increased risk of HAV infection in WWs. METHODS: The prevalence of antibodies to HAV in 869 WWs was compared to 311 other subjects and analysed to detect the main potentially confounding variables. RESULTS: Univariate analysis demonstrated that occupational exposure to sewage was not significantly associated with the prevalence of anti-HAV(+). The anti-HAV(+) prevalence was strongly associated with age and shellfish consumption (P < 0.05) when the subcategories of workers were examined separately (WWs and control group) and jointly. In the logistic regression model, a significant association between anti-HAV(+) prevalence and duration of employment (P < 0.05) was found. The interaction term (age x duration of employment) was significant (P < 0.001) when included in the logistic model. CONCLUSIONS: This study shows that working in a wastewater treatment plant does not seem to be related to a greater prevalence of antibodies to hepatitis A. Moreover, the relative risk of HAV infection among WWs seems to be correlated with low anti-HAV(+) prevalence in the general population. PMID- 19561055 TI - Imaging in patients with acute abdominal pain. PMID- 19561057 TI - Survey of rice proteins interacting with OsFCA and OsFY proteins which are homologous to the Arabidopsis flowering time proteins, FCA and FY. AB - The FCA protein is involved in controlling flowering time and plays more general roles in RNA-mediated chromatin silencing in Arabidopsis. It contains two RNA binding domains and a WW domain. The FCA protein interacts with FY, a polyadenylation factor, via its WW domain. We previously characterized a rice gene, OsFCA, which was homologous to FCA. Here, we found that the OsFCA protein could interact through its WW domain with the following proteins: OsFY, a protein containing a CID domain present in RNA-processing factors such as Pcf11 and Nrd1; a protein similar to splicing factor SF1; a protein similar to FUSE splicing factor; and OsMADS8. The FY protein is associated with the 3' end processing machinery in Arabidopsis. Thus, we examined interactions between OsFY and the rice homologs (OsCstF-50, -64 and -77) of the AtCstF-50, -64 and -77 proteins. We found that OsFY could bind OsCstF50, whereas the OsCstF77 protein could bridge the interaction between OsCstF50 and OsCstF64. Taken together, our data suggest that OsFCA could interact with several proteins other than OsFY through its WW domain and may play several roles in rice. PMID- 19561056 TI - Imaging strategies for detection of urgent conditions in patients with acute abdominal pain: diagnostic accuracy study. AB - OBJECTIVE: To identify an optimal imaging strategy for the accurate detection of urgent conditions in patients with acute abdominal pain. DESIGN: Fully paired multicentre diagnostic accuracy study with prospective data collection. SETTING: Emergency departments of two university hospitals and four large teaching hospitals in the Netherlands. PARTICIPANTS: 1021 patients with non-traumatic abdominal pain of >2 hours' and <5 days' duration. Exclusion criteria were discharge from the emergency department with no imaging considered warranted by the treating physician, pregnancy, and haemorrhagic shock. INTERVENTION: All patients had plain radiographs (upright chest and supine abdominal), ultrasonography, and computed tomography (CT) after clinical and laboratory examination. A panel of experienced physicians assigned a final diagnosis after six months and classified the condition as urgent or non-urgent. MAIN OUTCOME MEASURES: Sensitivity and specificity for urgent conditions, percentage of missed cases and false positives, and exposure to radiation for single imaging strategies, conditional imaging strategies (CT after initial ultrasonography), and strategies driven by body mass index and age or by location of pain. RESULTS: 661 (65%) patients had a final diagnosis classified as urgent. The initial clinical diagnosis resulted in many false positive urgent diagnoses, which were significantly reduced after ultrasonography or CT. CT detected more urgent diagnoses than did ultrasonography: sensitivity was 89% (95% confidence interval 87% to 92%) for CT and 70% (67% to 74%) for ultrasonography (P<0.001). A conditional strategy with CT only after negative or inconclusive ultrasonography yielded the highest sensitivity, missing only 6% of urgent cases. With this strategy, only 49% (46% to 52%) of patients would have CT. Alternative strategies guided by body mass index, age, or location of the pain would all result in a loss of sensitivity. CONCLUSION: Although CT is the most sensitive imaging investigation for detecting urgent conditions in patients with abdominal pain, using ultrasonography first and CT only in those with negative or inconclusive ultrasonography results in the best sensitivity and lowers exposure to radiation. PMID- 19561059 TI - The spectrum of sanity and insanity. PMID- 19561058 TI - Gray matter deficits, mismatch negativity, and outcomes in schizophrenia. AB - Reduced mismatch negativity (MMN) in response to auditory change is a well established finding in schizophrenia and has been shown to be correlated with impaired daily functioning, rather than with hallmark signs and symptoms of the disorder. In this study, we investigated (1) whether the relationship between reduced MMN and impaired daily functioning is mediated by cortical volume loss in temporal and frontal brain regions in schizophrenia and (2) whether this relationship varies with the type of auditory deviant generating MMN. MMN in response to duration, frequency, and intensity deviants was recorded from 18 schizophrenia subjects and 18 pairwise age- and gender-matched healthy subjects. Patients' levels of global functioning were rated on the Social and Occupational Functioning Assessment Scale. High-resolution structural magnetic resonance scans were acquired to generate average cerebral cortex and temporal lobe models using cortical pattern matching. This technique allows accurate statistical comparison and averaging of cortical measures across subjects, despite wide variations in gyral patterns. MMN amplitude was reduced in schizophrenia patients and correlated with their impaired day-to-day function level. Only in patients, bilateral gray matter reduction in Heschl's gyrus, as well as motor and executive regions of the frontal cortex, correlated with reduced MMN amplitude in response to frequency deviants, while reduced gray matter in right Heschl's gyrus also correlated with reduced MMN to duration deviants. Our findings further support the importance of MMN reduction in schizophrenia by linking frontotemporal cerebral gray matter pathology to an automatically generated event-related potential index of daily functioning. PMID- 19561060 TI - Spatial distribution of deep sulcal landmarks and hemispherical asymmetry on the cortical surface. AB - The locally deepest regions of major sulci, the sulcal pits, are thought to be the first cortical folds to develop and are closely related to functional areas. We examined the spatial distribution of sulcal pits across the entire cortical region, and assessed the hemispheric asymmetry in their frequency and distribution in a large group of normal adult brains. We automatically extracted sulcal pits from magnetic resonance imaging data using surface-based methods and constructed a group map from 148 subjects. The spatial distribution of the sulcal pits was relatively invariant between individuals, showing high frequency and density in specific focal areas. The left and right sulcal pits were spatially covariant in the regions of the earliest developed sulci. The sulcal pits with great spatial invariance appear to be useful as stable anatomical landmarks. We showed the most significant asymmetry in the frequency and spatial variance of sulcal pits in the superior temporal sulcus, which might be related to the lateralization of language function to the left hemisphere, developing more consistently and strongly than for the right. Our analyses support previous empirical and theoretical studies, and provide additional insights concerning the anatomical and functional development of the brain. PMID- 19561061 TI - Expectancy constraints in degraded speech modulate the language comprehension network. AB - In speech comprehension, the processing of auditory information and linguistic context are mutually dependent. This functional magnetic resonance imaging study examines how semantic expectancy ("cloze probability") in variably intelligible sentences ("noise vocoding") modulates the brain bases of comprehension. First, intelligibility-modulated activation along the superior temporal sulci (STS) was extended anteriorly and posteriorly in low-cloze sentences (e.g., "she weighs the flour") but restricted to a mid-superior temporal gyrus/STS area in more predictable high-cloze sentences (e.g., "she sifts the flour"). Second, the degree of left inferior frontal gyrus (IFG) (Brodmann's area 44) involvement in processing low-cloze constructions was proportional to increasing intelligibility. Left inferior parietal cortex (IPC; angular gyrus) activation accompanied successful speech comprehension that derived either from increased signal quality or from semantic facilitation. The results show that successful decoding of speech in auditory cortex areas regulates language-specific computation (left IFG and IPC). In return, semantic expectancy can constrain these speech-decoding processes, with fewer neural resources being allocated to highly predictable sentences. These findings offer an important contribution toward the understanding of the functional neuroanatomy in speech comprehension. PMID- 19561062 TI - Reading aloud boosts connectivity through the putamen. AB - Functional neuroimaging and lesion studies have frequently reported thalamic and putamen activation during reading and speech production. However, it is currently unknown how activity in these structures interacts with that in other reading and speech production areas. This study investigates how reading aloud modulates the neuronal interactions between visual recognition and articulatory areas, when both the putamen and thalamus are explicitly included. Using dynamic causal modeling in skilled readers who were reading regularly spelled English words, we compared 27 possible pathways that might connect the ventral anterior occipito temporal sulcus (aOT) to articulatory areas in the precentral cortex (PrC). We focused on whether the neuronal interactions within these pathways were increased by reading relative to picture naming and other visual and articulatory control conditions. The results provide strong evidence that reading boosts the aOT-PrC pathway via the putamen but not the thalamus. However, the putamen pathway was not exclusive because there was also evidence for another reading pathway that did not involve either the putamen or the thalamus. We conclude that the putamen plays a special role in reading but this is likely to vary with individual reading preferences and strategies. PMID- 19561063 TI - Output properties and organization of the forelimb representation of motor areas on the lateral aspect of the hemisphere in rhesus macaques. AB - Motor output capabilities of the forelimb representation of dorsal motor area (PMd) and ventral motor area (PMv) were compared with primary motor cortex (M1) in terms of latency, strength, sign, and distribution of effects. Stimulus triggered averages (60 microA) of electromyographic activity collected from 24 forelimb muscles were computed at 314 tracks in 2 monkeys trained to perform a reach-to-grasp task. The onset latency and magnitude of facilitation effects from PMd and PMv were significantly longer and 7- to 9-fold weaker than those from M1. Proximal muscles were predominantly represented in PMd and PMv. A joint-dependent flexor or extensor preference was also present. Distal and proximal muscle representations were intermingled in PMd and PMv. A gradual increase in latency and decrease in magnitude of effects were observed in moving from M1 surface sites toward more anterior sites in PMd. For many muscles, segregated areas producing suppression effects were found along the medial portion of PMd and adjacent M1. Although some facilitation effects from PMd and PMv had onset latencies as short as those from M1 in the same muscle, suggesting equal direct linkage, the vast majority had properties consistent with a more indirect linkage to motoneurons either through corticocortical connections with M1 and/or interneuronal linkages in the spinal cord. PMID- 19561064 TI - Cigarette smoking and pancreatic cancer: a pooled analysis from the pancreatic cancer cohort consortium. AB - Smoking is an established risk factor for pancreatic cancer; however, detailed examination of the association of smoking intensity, smoking duration, and cumulative smoking dose with pancreatic cancer is limited. The authors analyzed pooled data from the international Pancreatic Cancer Cohort Consortium nested case-control study (1,481 cases, 1,539 controls). Odds ratios and 95% confidence intervals were calculated by using unconditional logistic regression. Smoking intensity effects were examined with an excess odds ratio model that was linear in pack-years and exponential in cigarettes smoked per day and its square. When compared with never smokers, current smokers had a significantly elevated risk (odds ratio (OR) = 1.77, 95% confidence interval (CI): 1.38, 2.26). Risk increased significantly with greater intensity (> or =30 cigarettes/day: OR = 1.75, 95% CI: 1.27, 2.42), duration (> or =50 years: OR = 2.13, 95% CI: 1.25, 3.62), and cumulative smoking dose (> or =40 pack-years: OR = 1.78, 95% CI: 1.35, 2.34). Risk more than 15 years after smoking cessation was similar to that for never smokers. Estimates of excess odds ratio per pack-year declined with increasing intensity, suggesting greater risk for total exposure delivered at lower intensity for longer duration than for higher intensity for shorter duration. This finding and the decline in risk after smoking cessation suggest that smoking has a late-stage effect on pancreatic carcinogenesis. PMID- 19561065 TI - Mortality rates among trichlorophenol workers with exposure to 2,3,7,8 tetrachlorodibenzo-p-dioxin. AB - The authors examined 1,615 workers exposed to dioxins in trichlorophenol production in Midland, Michigan, to determine if there were increased mortality rates from exposure. Historical dioxin levels were estimated by a serum survey of workers. Vital status was followed from 1942 to 2003, and cause-specific death rates and trends with exposure were evaluated. All cancers combined (standardized mortality ratio (SMR) = 1.0, 95% confidence interval (CI): 0.8, 1.1), lung cancers (SMR = 0.7, 95% CI: 0.5, 0.9), and nonmalignant respiratory disease (SMR = 0.8, 95% CI: 0.6, 1.0) were at or below expected levels. Observed deaths for leukemia (SMR = 1.9, 95% CI: 1.0, 3.2), non-Hodgkin lymphoma (SMR = 1.3, 95% CI: 0.6, 2.5), diabetes (SMR = 1.1, 95% CI: 0.6, 1.8), and ischemic heart disease (SMR = 1.1, 95% CI: 0.9, 1.2) were slightly greater than expected. No trend was observed with exposure for these causes of death. However, for 4 deaths of soft tissue sarcoma (SMR = 4.1, 95% CI: 1.1, 10.5), the mortality rates increased with exposure. The small number of deaths and the uncertainty in both diagnosis and nosology coding make interpretation of this finding tenuous. With the exception of soft tissue sarcoma, the authors found little evidence of increased disease risk from exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin. PMID- 19561066 TI - Direct interdomain interactions can mediate allosterism in the thyroid receptor. AB - The thyroid (TR) and retinoid X receptors (RXR) belong to the nuclear receptor (NR) superfamily of ligand-mediated transcription factors. At the molecular level, TR activity is specifically modulated by interactions with the ligand 3,3',5 triiodo-l-thyronine (T3), RXR, DNA, and co-activators such as SRC1, occurring in concert or sequentially. Although binding sites for DNA and coregulators such as SRC1 are distinct and at distal regions of these receptors, cell-based and EMSA studies have suggested that these molecules can regulate binding of each other to the receptor. We present evidence of direct, DNA dependent, communication between the DNA and ligand binding domains (DBD and LBD) that can allosterically regulate interactions with SRC1 and DNA, respectively, using isothermal titration calorimetry (ITC) and cell-based assays. Additionally, we note that interdomain communication is affected by RXR in RXR:TR. We also noticed a DNA-dependent cross-talk between RXR and TR within RXR:TR. Finally, we suggest that differences in transactivation on different TRE may be the consequence of different affinities between TRE and RXR:TR. PMID- 19561067 TI - Palmitoylation of interferon-alpha (IFN-alpha) receptor subunit IFNAR1 is required for the activation of Stat1 and Stat2 by IFN-alpha. AB - Type I interferons (IFNs) bind IFNAR receptors and activate Jak kinases and Stat transcription factors to stimulate the transcription of genes downstream from IFN stimulated response elements. In this study, we analyze the role of protein palmitoylation, a reversible post-translational lipid modification, in the functional properties of IFNAR. We report that pharmacological inhibition of protein palmitoylation results in severe defects of IFN receptor endocytosis and signaling. We generated mutants of the IFNAR1 subunit of the type I IFN receptor, in which each or both of the two cysteines present in the cytoplasmic domain are replaced by alanines. We show that cysteine 463 of IFNAR1, the more proximal of the two cytoplasmic cysteines, is palmitoylated. A thorough microscopic and biochemical analysis of the palmitoylation-deficient IFNAR1 mutant revealed that IFNAR1 palmitoylation is not required for receptor endocytosis, intracellular distribution, or stability at the cell surface. However, the lack of IFNAR1 palmitoylation affects selectively the activation of Stat2, which results in a lack of efficient Stat1 activation and nuclear translocation and IFN-alpha activated gene transcription. Thus, receptor palmitoylation is a previously undescribed mechanism of regulating signaling activity by type I IFNs in the Jak/Stat pathway. PMID- 19561068 TI - Deorphanization of GPR109B as a receptor for the beta-oxidation intermediate 3-OH octanoic acid and its role in the regulation of lipolysis. AB - The orphan G-protein-coupled receptor GPR109B is the result of a recent gene duplication of the nicotinic acid and ketone body receptor GPR109A being found in humans but not in rodents. Like GPR109A, GPR109B is predominantly expressed in adipocytes and is supposed to mediate antilipolytic effects. Here we show that GPR109B serves as a receptor for the beta-oxidation intermediate 3-OH-octanoic acid, which has antilipolytic activity on human but not on murine adipocytes. GPR109B is coupled to Gi-type G-proteins and is activated by 2- and 3-OH-octanoic acid with EC50 values of about 4 and 8 microM, respectively. Interestingly, 3-OH octanoic acid plasma concentrations reach micromolar concentrations under conditions of increased beta-oxidation rates, like in diabetic ketoacidosis or under a ketogenic diet. These data suggest that the ligand receptor pair 3-OH octanoic acid/GPR109B mediates in humans a negative feedback regulation of adipocyte lipolysis to counteract prolipolytic influences under conditions of physiological or pathological increases in beta-oxidation rates. PMID- 19561069 TI - Mammalian cytochrome P450 enzymes catalyze the phenol-coupling step in endogenous morphine biosynthesis. AB - A cytochrome P450 (P450) enzyme in porcine liver that catalyzed the phenol coupling reaction of the substrate (R)-reticuline to salutaridine was previously purified to homogeneity (Amann, T., Roos, P. H., Huh, H., and Zenk, M. H. (1995) Heterocycles 40, 425-440). This reaction was found to be catalyzed by human P450s 2D6 and 3A4 in the presence of (R)-reticuline and NADPH to yield not a single product, but rather (-)-isoboldine, (-)-corytuberine, (+)-pallidine, and salutaridine, the para-ortho coupled established precursor of morphine in the poppy plant and most likely also in mammals. (S)-Reticuline, a substrate of both P450 enzymes, yielded the phenol-coupled alkaloids (+)-isoboldine, (+) corytuberine, (-)-pallidine, and sinoacutine; none of these serve as a morphine precursor. Catalytic efficiencies were similar for P450 2D6 and P450 3A4 in the presence of cytochrome b(5) with (R)-reticuline as substrate. The mechanism of phenol coupling is not yet established; however, we favor a single cycle of iron oxidation to yield salutaridine and the three other alkaloids from (R) reticuline. The total yield of salutaridine formed can supply the 10 nm concentration of morphine found in human neuroblastoma cell cultures and in brain tissues of mice. PMID- 19561070 TI - PRC1 cooperates with CLASP1 to organize central spindle plasticity in mitosis. AB - During cell division, chromosome segregation is governed by the interaction of spindle microtubules with the kinetochore. A dramatic remodeling of interpolar microtubules into an organized central spindle between the separating chromatids is required for the initiation and execution of cytokinesis. Central spindle organization requires mitotic kinesins, microtubule-bundling protein PRC1, and Aurora B kinase complex. However, the precise role of PRC1 in central spindle organization has remained elusive. Here we show that PRC1 recruits CLASP1 to the central spindle at early anaphase onset. CLASP1 belongs to a conserved microtubule-binding protein family that mediates the stabilization of overlapping microtubules of the central spindle. PRC1 physically interacts with CLASP1 and specifies its localization to the central spindle. Repression of CLASP1 leads to sister-chromatid bridges and depolymerization of spindle midzone microtubules. Disruption of PRC1-CLASP1 interaction by a membrane-permeable peptide abrogates accurate chromosome segregation, resulting in sister chromatid bridges. These findings reveal a key role for the PRC1-CLASP1 interaction in achieving a stable anti-parallel microtubule organization essential for faithful chromosome segregation. We propose that PRC1 forms a link between stabilization of CLASP1 association with central spindle microtubules and anti-parallel microtubule elongation. PMID- 19561071 TI - Formation of the stable structural analog of ADP-sensitive phosphoenzyme of Ca2+ ATPase with occluded Ca2+ by beryllium fluoride: structural changes during phosphorylation and isomerization. AB - As a stable analog for ADP-sensitive phosphorylated intermediate of sarcoplasmic reticulum Ca(2+)-ATPase E1PCa(2).Mg, a complex of E1Ca(2).BeF(x), was successfully developed by addition of beryllium fluoride and Mg(2+) to the Ca(2+) bound state, E1Ca(2). In E1Ca(2).BeF(x), most probably E1Ca(2).BeF(3)(-), two Ca(2+) are occluded at high affinity transport sites, its formation required Mg(2+) binding at the catalytic site, and ADP decomposed it to E1Ca(2), as in E1PCa(2).Mg. Organization of cytoplasmic domains in E1Ca(2).BeF(x) was revealed to be intermediate between those in E1Ca(2).AlF(4)(-) ADP (transition state of E1PCa(2) formation) and E2.BeF(3)(-).(ADP-insensitive phosphorylated intermediate E2P.Mg). Trinitrophenyl-AMP (TNP-AMP) formed a very fluorescent (superfluorescent) complex with E1Ca(2).BeF(x) in contrast to no superfluorescence of TNP-AMP bound to E1Ca(2).AlF(x). E1Ca(2).BeF(x) with bound TNP-AMP slowly decayed to E1Ca(2), being distinct from the superfluorescent complex of TNP-AMP with E2.BeF(3)(-), which was stable. Tryptophan fluorescence revealed that the transmembrane structure of E1Ca(2).BeF(x) mimics E1PCa(2).Mg, and between those of E1Ca(2).AlF(4)(-).ADP and E2.BeF(3)(-). E1Ca(2).BeF(x) at low 50-100 microm Ca(2+) was converted slowly to E2.BeF(3)(-) releasing Ca(2+), mimicking E1PCa(2).Mg --> E2P.Mg + 2Ca(2+). Ca(2+) replacement of Mg(2+) at the catalytic site at approximately millimolar high Ca(2+) decomposed E1Ca(2).BeF(x) to E1Ca(2). Notably, E1Ca(2).BeF(x) was perfectly stabilized for at least 12 days by 0.7 mm lumenal Ca(2+) with 15 mm Mg(2+). Also, stable E1Ca(2).BeF(x) was produced from E2.BeF(3)(-) at 0.7 mm lumenal Ca(2+) by binding two Ca(2+) to lumenally oriented low affinity transport sites, as mimicking the reverse conversion E2P. Mg + 2Ca(2+) --> E1PCa(2).Mg. PMID- 19561072 TI - alpha-Helical domains promote translocation of intrinsically disordered polypeptides into the endoplasmic reticulum. AB - Co-translational import into the endoplasmic reticulum (ER) is primarily controlled by N-terminal signal sequences that mediate targeting of the ribosome nascent chain complex to the Sec61/translocon and initiate the translocation process. Here we show that after targeting to the translocon the secondary structure of the nascent polypeptide chain can significantly modulate translocation efficiency. ER-targeted polypeptides dominated by unstructured domains failed to efficiently translocate into the ER lumen and were subjected to proteasomal degradation via a co-translocational/preemptive pathway. Productive ER import could be reinstated by increasing the amount of alpha-helical domains, whereas more effective ER signal sequences had only a minor effect on ER import efficiency of unstructured polypeptides. ER stress and overexpression of p58(IPK) promoted the co-translocational degradation pathway. Moreover polypeptides with unstructured domains at their N terminus were specifically targeted to proteasomal degradation under these conditions. Our study indicates that extended unstructured domains are signals to dispose ER-targeted proteins via a co translocational, preemptive quality control pathway. PMID- 19561074 TI - Regulation of phosphatidylinositol kinases and metabolism by Wnt3a and Dvl. AB - Wnt signaling plays important roles in various physiological and pathophysiological processes. The pathway that leads to beta-catenin stabilization is initiated by Wnt binding to its cell surface receptors, which induces the formation of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)) via activation of phosphatidylinositol 4-phosphate 5-kinase (PIP5K) type I. Here, we show that Wnt also stimulated the production of phosphatidylinositol 4 phosphate (PtdIns(4)P), which depended on Frizzled (Fz), Dishevelled (Dvl), and phosphatidylinositol 4-kinase (PI4K) type II alpha in HEK293T cells. Dvl directly interacted with and activated PI4KII alpha by increasing its V(max) for ATP and PtdIns. In addition, Dvl regulated PI4KII alpha and PIP5KI via different domains. Moreover, Dvl, PI4KII alpha, and PIP5KI appeared to form a ternary complex upon Wnt3a stimulation. This complex may allow efficient production of PtdIns(4,5)P(2) from PtdIns, which is far more abundant than PtdIns(4)P in cells. Therefore, this study provides new insights into the mechanism by which Wnt3a regulates the production of PtdIns(4,5)P(2). PMID- 19561073 TI - A novel family of Apicomplexan glideosome-associated proteins with an inner membrane-anchoring role. AB - The phylum Apicomplexa are a group of obligate intracellular parasites responsible for a wide range of important diseases. Central to the lifecycle of these unicellular parasites is their ability to migrate through animal tissue and invade target host cells. Apicomplexan movement is generated by a unique system of gliding motility in which substrate adhesins and invasion-related proteins are pulled across the plasma membrane by an underlying actin-myosin motor. The myosins of this motor are inserted into a dual membrane layer called the inner membrane complex (IMC) that is sandwiched between the plasma membrane and an underlying cytoskeletal basket. Central to our understanding of gliding motility is the characterization of proteins residing within the IMC, but to date only a few proteins are known. We report here a novel family of six-pass transmembrane proteins, termed the GAPM family, which are highly conserved and specific to Apicomplexa. In Plasmodium falciparum and Toxoplasma gondii the GAPMs localize to the IMC where they form highly SDS-resistant oligomeric complexes. The GAPMs co purify with the cytoskeletal alveolin proteins and also to some degree with the actin-myosin motor itself. Hence, these proteins are strong candidates for an IMC anchoring role, either directly or indirectly tethering the motor to the cytoskeleton. PMID- 19561075 TI - Smad6 inhibits the transcriptional activity of Tbx6 by mediating its degradation. AB - Members of the bone morphogenetic protein (BMP) and T-box gene families play several critical roles in the early embryonic development and tissue homeostasis. Although BMP proteins are the upstream regulators of T-box genes, few studies have investigated the molecular mechanisms between these two protein families. Here, we report that Tbx6 interacts directly with Smad6, an inhibitory Smad that antagonizes the BMP signal. This interaction is mediated through the Mad homology 2 (MH2) domain of Smad6 and residues 90-180 of Tbx6. We demonstrate that Smad6 facilitates the degradation of Tbx6 protein through recruitment of Smurf1, a ubiquitin E3 ligase. Consequently, Smad6 reduces Tbx6-mediated Myf-5 gene activation. Furthermore, specific knockdown of endogenous Smad6 and Smurf1 by small interfering RNA increases the protein levels of Tbx6 and enhance the expression of Tbx6 target genes. Collectively, these findings reveal that Smad6 serves as a critical mediator of BMP signal via a functional interaction with Tbx6, thus regulating the activation of Tbx6 downstream genes during cell differentiation. PMID- 19561076 TI - Single-molecule study on the temperature-sensitive reaction of F1-ATPase with a hybrid F1 carrying a single beta(E190D). AB - F(1)-ATPase is a rotary molecular motor in which the gamma-subunit rotates against the alpha(3)beta(3) cylinder. The unitary gamma-rotation is a 120 degrees step comprising 80 and 40 degrees substeps, each of these initiated by ATP binding and ADP release and by ATP hydrolysis and inorganic phosphate release, respectively. In our previous study on gamma-rotation at low temperatures, a highly temperature-sensitive (TS) reaction step of F(1)-ATPase from thermophilic Bacillus PS3 was found below 9 degrees C as an intervening pause before the 80 degrees substep at the same angle for ATP binding and ADP release. However, it remains unclear as to which reaction step the TS reaction corresponds. In this study, we found that the mutant F(1)(beta E190D) from thermophilic Bacillus PS3 showed a clear pause of the TS reaction below 18 degrees C. In an attempt to identify the catalytic state of the TS reaction, the rotation of the hybrid F(1), carrying a single copy of beta E190D, was observed at 18 degrees C. The hybrid F(1) showed a pause of the TS reaction at the same angle as for the ATP binding of the incorporated beta E190D, although kinetic analysis revealed that the TS reaction is not the ATP binding step. These findings suggest that the TS reaction is a structural rearrangement of beta before or after ATP binding. PMID- 19561077 TI - Identification of the determinant conferring permissive substrate usage in the telomere resolvase, ResT. AB - Linear genome stability requires specialized telomere replication and protection mechanisms. A common solution to this problem in non-eukaryotes is the formation of hairpin telomeres by telomere resolvases (also known as protelomerases). These enzymes perform a two-step transesterification on replication intermediates to generate hairpin telomeres using an active site similar to that of tyrosine recombinases and type IB topoisomerases. Unlike phage telomere resolvases, the telomere resolvase from the Lyme disease pathogen Borrelia burgdorferi (ResT) is a permissive enzyme that resolves several types of telomere in vitro. However, the ResT region and residues mediating permissive substrate usage have not been identified. The relapsing fever Borrelia hermsii ResT exhibits a more restricted substrate usage pattern than B. burgdorferi ResT and cannot efficiently resolve a Type 2 telomere. In this study, we determined that all relapsing fever ResTs process Type 2 telomeres inefficiently. Using a library of chimeric and mutant B. hermsii/B. burgdorferi ResTs, we mapped the determinants in B. burgdorferi ResT conferring the ability to resolve multiple Type 2 telomeres. Type 2 telomere resolution was dependent on a single proline in the ResT catalytic region that was conserved in all Lyme disease but not relapsing fever ResTs and that is part of a 2-amino acid insertion absent from phage telomere resolvase sequences. The identification of a permissive substrate usage determinant explains the ability of B. burgdorferi ResT to process the 19 unique telomeres found in its segmented genome and will aid further studies on the structure and function of this essential enzyme. PMID- 19561078 TI - Knockdown of ASIC1 and epithelial sodium channel subunits inhibits glioblastoma whole cell current and cell migration. AB - High grade gliomas such as glioblastoma multiforme express multiple members of the epithelial sodium channel (ENaC)/Degenerin family, characteristically displaying a basally active amiloride-sensitive cation current not seen in normal human astrocytes or lower grade gliomas. Using quantitative real time PCR, we have shown higher expression of ASIC1, alphaENaC, and gammaENaC in D54-MG human glioblastoma multiforme cells compared with primary human astrocytes. We hypothesize that this glioma current is mediated by a hybrid channel composed of a mixture of ENaC and acid-sensing ion channel (ASIC) subunits. To test this hypothesis we made dominant negative cDNAs for ASIC1, alphaENaC, gammaENaC, and deltaENaC. D54-MG cells transfected with the dominant negative constructs for ASIC1, alphaENaC, or gammaENaC showed reduced protein expression and a significant reduction in the amiloride-sensitive whole cell current as compared with untransfected D54-MG cells. Knocking down alphaENaC or gammaENaC also abolished the high P(K)(+)/P(Na)(+) of D54-MG cells. Knocking down deltaENaC in D54-MG cells reduced deltaENaC protein expression but had no effect on either the whole cell current or K(+) permeability. Using co-immunoprecipitation we show interactions between ASIC1, alphaENaC, and gammaENaC, consistent with these subunits interacting with each other to form an ion channel in glioma cells. We also found a significant inhibition of D54-MG cell migration after ASIC1, alphaENaC, or gammaENaC knockdown, consistent with the hypothesis that ENaC/Degenerin subunits play an important role in glioma cell biology. PMID- 19561079 TI - The thioxotriazole copper(II) complex A0 induces endoplasmic reticulum stress and paraptotic death in human cancer cells. AB - The copper(II) complex A0 induces a type of non-apoptotic cell death also known as paraptosis. Paraptosis involves extensive endoplasmic reticulum vacuolization in the absence of caspase activation. A wide panel of human cancer cell lines was used to demonstrate differences in cytotoxicity by the paraptosis-inducing drug A0 and the metal-based pro-apoptotic drug cisplatin. Gene expression profiling of the human fibrosarcoma HT1080 cells showed that, while cisplatin induced p53 targets, A0 up-regulated genes involved in the unfolded protein response (UPR) and response to heavy metals. The cytotoxic effects of A0 were associated with inhibition of the ubiquitin-proteasome system and accumulation of ubiquitinylated proteins, in a manner dependent on protein synthesis. Cycloheximide inhibited the accumulation of ubiquitinylated proteins and hampered A0-induced cell death process. The occurrence of the UPR during A0-induced death process was shown by the increased abundance of spliced XBP1 mRNA, transient eIF2alpha phosphorylation, and a series of downstream events, including attenuation of global protein synthesis and increased expression of ATF4, CHOP, BIP, and GADD34. Mouse embryonic fibroblasts expressing a mutant eIF2alpha, which could not be phosphorylated, were more resistant to A0 than wild type cells, pointing to a pro death role of eIF2alpha phosphorylation. A0 may thus represent the prototypical member of a new class of compounds that cause paraptotic cell death via mechanisms involving eIF2alpha phosphorylation and the UPR. PMID- 19561080 TI - Role of conserved salt bridges in homeodomain stability and DNA binding. AB - The sequence information available for homeodomains reveals that salt bridges connecting pairs 19/30, 31/42, and 17/52 are frequent, whereas aliphatic residues at these sites are rare and mainly restricted to proteins from homeotherms. We have analyzed the influence of salt and hydrophobic bridges at these sites on the stability and DNA binding properties of human Hesx-1 homeodomain. Regarding the protein stability, our analysis shows that hydrophobic side chains are clearly preferred at positions 19/30 and 31/42. This stabilizing influence results from the more favorable packing of the aliphatic side chains with the protein core, as illustrated by the three-dimensional solution structure of a thermostable variant, herein reported. In contrast only polar side chains seem to be tolerated at positions 17/52. Interestingly, despite the significant influence of pairs 19/30 and 31/42 on the stability of the homeodomain, their effect on DNA binding ranges from modest to negligible. The observed lack of correlation between binding strength and conformational stability in the analyzed variants suggests that salt/hydrophobic bridges at these specific positions might have been employed by evolution to independently modulate both properties. PMID- 19561081 TI - Suicidal membrane repair regulates phosphatidylserine externalization during apoptosis. AB - One of the hallmarks of apoptosis is the redistribution of phosphatidylserine (PS) from the inner-to-outer plasma membrane (PM) leaflet, where it functions as a ligand for phagocyte recognition and the suppression of inflammatory responses. The mechanism by which apoptotic cells externalize PS has been assumed to involve "scramblases" that randomize phospholipids across the PM bilayer. These putative activities, however, have not been unequivocally proven to be responsible for the redistribution of lipids. Because elevated cytosolic Ca(2+) is critical to this process and is also required for activation of lysosome-PM fusion during membrane repair, we hypothesized that apoptosis could activate a "pseudo"-membrane repair response that results in the fusion of lysosomes with the PM. Using a membrane specific probe that labels endosomes and lysosomes and fluorescein-labeled annexin 5 that labels PS, we show that the appearance of PS at the cell surface during apoptosis is dependent on the fusion of lysosomes with the PM, a process that is inhibited with the lysosomotrophe, chloroquine. We demonstrate that apoptotic cells evoke a persistent pseudo-membrane repair response that likely redistributes lysosomal-derived PS to the PM outer leaflet that leads to membrane expansion and the formation of apoptotic blebs. Our data suggest that inhibition of lysosome-PM fusion-dependent redistribution of PS that occurs as a result of chemotherapy- and radiotherapy-induced apoptosis will prevent PS-dependent anti inflammatory responses that preclude the development of tumor- and patient specific immune responses. PMID- 19561082 TI - Mycobacterial phosphatidylinositol mannosides negatively regulate host Toll-like receptor 4, MyD88-dependent proinflammatory cytokines, and TRIF-dependent co stimulatory molecule expression. AB - Mycobacterium tuberculosis modulates host immune responses through proteins and complex glycolipids. Here, we report that the glycosylphosphatidylinositol anchor phosphatidyl-myo-inositol hexamannosides PIM(6) or PIM(2) exert potent anti inflammatory activities. PIM strongly inhibited the Toll-like receptor (TLR4) and myeloid differentiation protein 88 (MyD88)-mediated release of NO, cytokines, and chemokines, including tumor necrosis factor (TNF), interleukin 12 (IL-12) p40, IL 6, keratinocyte-derived chemokine, and also IL-10 by lipopolysaccharide (LPS) activated macrophages. This effect was independent of the presence of TLR2. PIM also reduced the LPS-induced MyD88-independent, TIR domain-containing adaptor protein inducing interferon beta (TRIF)-mediated expression of co-stimulatory receptors. PIM inhibited LPS/TLR4-induced NFkappaB translocation. Synthetic PIM(1) and a PIM(2) mimetic recapitulated these in vitro activities and inhibited endotoxin-induced airway inflammation, TNF and keratinocyte-derived chemokine secretion, and neutrophil recruitment in vivo. Mannosyl, two acyl chains, and phosphatidyl residues are essential for PIM anti-inflammatory activity, whereas the inosityl moiety is dispensable. Therefore, PIM exert potent antiinflammatory effects both in vitro and in vivo that may contribute to the strategy developed by mycobacteria for repressing the host innate immunity, and synthetic PIM analogs represent powerful anti-inflammatory leads. PMID- 19561083 TI - The mechanism of CSF-1-induced Wiskott-Aldrich syndrome protein activation in vivo: a role for phosphatidylinositol 3-kinase and Cdc42. AB - A role for Wiskott-Aldrich syndrome protein (WASP) in chemotaxis to various agents has been demonstrated in monocyte-derived cell types. Although WASP has been shown to be activated by multiple mechanisms in vitro, it is unclear how WASP is regulated in vivo. A WASP biosensor (WASPbs), which uses intramolecular fluorescence resonance energy transfer to report WASP activation in vivo, was constructed, and following transfection of macrophages, activation of WASPbs upon treatment with colony-stimulating factor-1 (CSF-1) was detected globally as early as 30 s and remained localized to protrusive regions at later time points. Similar results were obtained when endogenous WASP activation was determined using conformation-sensitive antibodies. In vivo CSF-1-induced WASP activation was fully Cdc42-dependent. Activation of WASP in response to treatment with CSF-1 was also shown to be phosphatidylinositol 3-kinase-dependent. However, treatment with the Src family kinase inhibitors PP2 or SU6656 or disruption of the major tyrosine phosphorylation site of WASPbs (Y291F mutation) did not reduce the level of CSF-1-induced WASP activation. Our results indicate that WASP activation downstream of CSF-1R is phosphatidylinositol 3-kinase- and Cdc42-dependent consistent with an involvement of these molecules in macrophage migration. However, although tyrosine phosphorylation of WASP has been proposed to stimulate WASP activity, we found no evidence to indicate that this occurs in vivo. PMID- 19561084 TI - Raptor binds the SAIN (Shc and IRS-1 NPXY binding) domain of insulin receptor substrate-1 (IRS-1) and regulates the phosphorylation of IRS-1 at Ser-636/639 by mTOR. AB - In normal physiological states mTOR phosphorylates and activates Akt. However, under diabetic-mimicking conditions mTOR inhibits phosphatidylinositol (PI) 3 kinase/Akt signaling by phosphorylating insulin receptor substrate-1 (IRS-1) at Ser-636/639. The molecular basis for the differential effect of mTOR signaling on Akt is poorly understood. Here, it has been shown that knockdown of mTOR, Raptor, and mLST8, but not Rictor and mSin1, suppresses insulin-stimulated phosphorylation of IRS-1 at Ser-636/639 and stabilizes IRS-1 after long term insulin stimulation. This phosphorylation depends on the PI 3-kinase/PDK1 axis but is Akt-independent. At the molecular level, Raptor binds the SAIN (Shc and IRS-1 NPXY binding) domain of IRS-1 and regulates the phosphorylation of IRS-1 at Ser-636/639 by mTOR. IRS-1 lacking the SAIN domain does not interact with Raptor, is not phosphorylated at Ser-636/639, and favorably interacts with PI 3-kinase. Overall, these data provide new insights in the molecular mechanisms by which mTORC1 inhibits PI 3-kinase/Akt signaling at the level of IRS-1 and suggest that mTOR signaling toward Akt is scaffold-dependent. PMID- 19561085 TI - Silencing of ErbB3/ErbB2 signaling by immunoglobulin-like Necl-2. AB - ErbB2 and ErbB3, members of the EGF receptor/ErbB family, form a heterodimer upon binding of a ligand, inducing the activation of Rac small G protein and Akt protein kinase for cell movement and survival, respectively. The enhanced ErbB3/ErbB2 signaling causes tumorigenesis, invasion, and metastasis. We found here that the ErbB3/ErbB2 signaling is regulated by immunoglobulin-like Necl-2, which is down-regulated in various cancer cells and serves as a tumor suppressor. The extracellular region of ErbB3, but not ErbB2, interacted in cis with that of Necl-2. This interaction reduced the ligand-induced, ErbB2-catalyzed tyrosine phosphorylation of ErbB3 and inhibited the consequent ErbB3-mediated activation of Rac and Akt, resulting in the inhibition of cancer cell movement and survival. These inhibitory effects of Necl-2 were mediated by the protein-tyrosine phosphatase PTPN13 which interacted with the cytoplasmic tail of Necl-2. We describe here this novel mechanism for silencing of the ErbB3/ErbB2 signaling by Necl-2. PMID- 19561086 TI - The small terminase, gp16, of bacteriophage T4 is a regulator of the DNA packaging motor. AB - Tailed bacteriophages and herpes viruses use powerful molecular motors to translocate DNA into a preassembled prohead and compact the DNA to near crystalline density. The phage T4 motor, a pentamer of 70-kDa large terminase, gp17, is the fastest and most powerful motor reported to date. gp17 has an ATPase activity that powers DNA translocation and a nuclease activity that cuts concatemeric DNA and generates the termini of viral genome. An 18-kDa small terminase, gp16, is also essential, but its role in DNA packaging is poorly understood. gp16 forms oligomers, most likely octamers, exhibits no enzymatic activities, but stimulates the gp17-ATPase activity, and inhibits the nuclease activity. Extensive mutational and biochemical analyses show that gp16 contains three domains, a central oligomerization domain, and N- and C-terminal domains that are essential for ATPase stimulation. Stimulation occurs not by nucleotide exchange or enhanced ATP binding but by triggering hydrolysis of gp17-bound ATP, a mechanism reminiscent of GTPase-activating proteins. gp16 does not have an arginine finger but its interaction with gp17 seems to position a gp17 arginine finger into the catalytic pocket. gp16 inhibits DNA translocation when gp17 is associated with the prohead. gp16 restricts gp17-nuclease such that the putative packaging initiation cut is made but random cutting is inhibited. These results suggest that the phage T4 packaging machine consists of a motor (gp17) and a regulator (gp16). The gp16 regulator is essential to coordinate the gp17 motor ATPase, translocase, and nuclease activities, otherwise it could be suicidal to the virus. PMID- 19561087 TI - A portable hot spot recognition loop transfers sequence preferences from APOBEC family members to activation-induced cytidine deaminase. AB - Enzymes of the AID/APOBEC family, characterized by the targeted deamination of cytosine to generate uracil within DNA, mediate numerous critical immune responses. One family member, activation-induced cytidine deaminase (AID), selectively introduces uracil into antibody variable and switch regions, promoting antibody diversity through somatic hypermutation or class switching. Other family members, including APOBEC3F and APOBEC3G, play an important role in retroviral defense by acting on viral reverse transcripts. These enzymes are distinguished from one another by targeting cytosine within different DNA sequence contexts; however, the reason for these differences is not known. Here, we report the identification of a recognition loop of 9-11 amino acids that contributes significantly to the distinct sequence motifs of individual family members. When this recognition loop is grafted from the donor APOBEC3F or 3G proteins into the acceptor scaffold of AID, the mutational signature of AID changes toward that of the donor proteins. These loop-graft mutants of AID provide useful tools for dissecting the biological impact of DNA sequence preferences upon generation of antibody diversity, and the results have implications for the evolution and specialization of the AID/APOBEC family. PMID- 19561088 TI - Intersubunit coupling in the pore of BK channels. AB - The structural basis underlying the gating of large conductance Ca(2+)-activated K(+) (BK) channels remains elusive. We found that substitution of Leu-312 in the S6 transmembrane segment of mSlo1 BK channels with hydrophilic amino acids of smaller side-chain volume favored the open state. The sensitivities of channels to calcium and voltage were modified by some mutations and completely abolished by others. Interpretation of the results in terms of an allosteric model suggests that the calcium-insensitive mutants greatly destabilize the closed relative to the open conformation and may also disrupt the allosteric coupling between Ca(2+) or voltage sensors and the gate. Some Phe-315 mutations also favor the open state, suggesting that Leu-312 and Phe-315 may interact in the closed state, forming a major energy barrier that the channel has to overcome to open. Homology modeling and molecular dynamic simulations further support that the side chain of Leu-312 can couple strongly with the aromatic ring of Phe-315 in neighboring subunits (L-F coupling) to maintain the channel closed. Additionally, single channel recordings indicate that the calcium-insensitive mutants, whose kinetics can be approximately characterized by a two-state closed-open (C-O) model, exhibit nearly 100% open probability under physiological conditions without alterations in single-channel conductance. These findings provide a basis for understanding the structure and gating of the BK channel pore. PMID- 19561090 TI - CGIN1: a retroviral contribution to mammalian genomes. AB - This study describes the origin and structural features of a mammalian gene, CGIN1 (Cousin of GIN1). CGIN1 proteins contain an NYN domain, retroviral RNase H and integrase domains, and a domain of unknown function (CGIN1 domain) that is also present in two other genes (N4BP1 and KIAA0323). We suggest that CGIN1 derives from the fusion of a KIAA0323-like gene with retroviral sequences, which occurred prior to the marsupial-eutherian split. Sequence and structural analyses indicate that the CGIN1 integrase domain is inactive but still retains the 3D folding observed in retroviral integrases. We hypothesize that CGIN1 may contribute to retroviral resistance in mammals by regulating the ubiquitination of viral proteins. PMID- 19561089 TI - B cell receptor-induced phosphorylation of Pyk2 and focal adhesion kinase involves integrins and the Rap GTPases and is required for B cell spreading. AB - Signaling by the B cell receptor (BCR) promotes integrin-mediated adhesion and cytoskeletal reorganization. This results in B cell spreading, which enhances the ability of B cells to bind antigens and become activated. Proline-rich tyrosine kinase (Pyk2) and focal adhesion kinase (FAK) are related cytoplasmic tyrosine kinases that regulate cell adhesion, cell morphology, and cell migration. In this report we show that BCR signaling and integrin signaling collaborate to induce the phosphorylation of Pyk2 and FAK on key tyrosine residues, a modification that increases the kinase activity of Pyk2 and FAK. Activation of the Rap GTPases is critical for BCR-induced integrin activation as well as for BCR- and integrin induced reorganization of the actin cytoskeleton. We now show that Rap activation is essential for BCR-induced phosphorylation of Pyk2 and for integrin-induced phosphorylation of Pyk2 and FAK. Moreover Rap-dependent phosphorylation of Pyk2 and FAK required an intact actin cytoskeleton as well as actin dynamics, suggesting that Rap regulates Pyk2 and FAK via its effects on the actin cytoskeleton. Importantly B cell spreading induced by BCR/integrin co-stimulation or by integrin engagement was inhibited by short hairpin RNA-mediated knockdown of either Pyk2 or FAK expression and by treatment with PF-431396, a chemical inhibitor that blocks the kinase activities of both Pyk2 and FAK. Thus Pyk2 and FAK are downstream targets of the Rap GTPases that play a key role in regulating B cell morphology. PMID- 19561091 TI - Lack of character displacement in the male recognition molecule, bindin, in Altantic sea urchins of the genus Echinometra. AB - Bindin, a protein involved in sea urchin sperm-egg recognition and adhesion, is under positive selection in genera with sympatric species but evolves neutrally in genera in which all species are allopatric. This pattern has led to suggestions that reinforcement may be the source of the observed selection. Reproductive character displacement, or increased divergence of reproductive characters in areas where closely related species overlap, is often a consequence of reinforcement and has been shown to be present in one Indo-Pacific species of the genus Echinometra. In the Atlantic species of the same genus, positive selection has been shown to act on bindin of Echinometra lucunter. To examine whether the source of this selection is reinforcement, we determined variation on the first exon of bindin in E. lucunter in the Caribbean, where it is sympatric with Echinometra viridis, and in the rest of the Atlantic, where E. viridis is absent. There was no differentiation between bindin sequences from the two geographic regions; similar levels of positive selection were found to be acting in both areas. The similarities were not due to gene flow; mitochondrial DNA from the two regions indicates that E. lucunter populations most likely originated in the Atlantic and have not exchanged genes with Caribbean populations for approximately 200,000 years. The lack of evidence of stronger selection on bindin of E. lucunter in areas of sympatry with its sister species suggests that the source of selection is not reinforcement. Processes acting within species, such as sexual selection, sperm competition, or sexual conflict, are more likely to be involved in the evolution of this molecule. PMID- 19561092 TI - Assessing computational methods for predicting protein stability upon mutation: good on average but not in the details. AB - Methods for protein modeling and design advanced rapidly in recent years. At the heart of these computational methods is an energy function that calculates the free energy of the system. Many of these functions were also developed to estimate the consequence of mutation on protein stability or binding affinity. In the current study, we chose six different methods that were previously reported as being able to predict the change in protein stability (DeltaDeltaG) upon mutation: CC/PBSA, EGAD, FoldX, I-Mutant2.0, Rosetta and Hunter. We evaluated their performance on a large set of 2156 single mutations, avoiding for each program the mutations used for training. The correlation coefficients between experimental and predicted DeltaDeltaG values were in the range of 0.59 for the best and 0.26 for the worst performing method. All the tested computational methods showed a correct trend in their predictions, but failed in providing the precise values. This is not due to lack in precision of the experimental data, which showed a correlation coefficient of 0.86 between different measurements. Combining the methods did not significantly improve prediction accuracy compared to a single method. These results suggest that there is still room for improvement, which is crucial if we want forcefields to perform better in their various tasks. PMID- 19561094 TI - Mycobacterium bovis bacillus Calmette-Guerin infection induces TLR2-dependent peroxisome proliferator-activated receptor gamma expression and activation: functions in inflammation, lipid metabolism, and pathogenesis. AB - Macrophages have important roles in both lipid metabolism and inflammation and are central to immunity to intracellular pathogens. Foam-like, lipid-laden macrophages are present during the course of mycobacterial infection and have recently been implicated in mycobacterial pathogenesis. In this study, we analyzed the molecular mechanisms underlying the formation of macrophage lipid bodies (lipid droplets) during Mycobacterium bovis bacillus Calmette-Guerin (BCG) infection, focusing on the role of the lipid-activated nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma). We found that BCG infection induced increased expression of PPARgamma that paralleled the augmented lipid body formation and PGE(2) synthesis in mouse peritoneal macrophages. BCG induced PPARgamma expression and lipid body formation were diminished in macrophages from TLR2-deficient mice, suggesting a key role for TLR2. The function of PPARgamma in modulating BCG infection was demonstrated by the capacity of the PPARgamma agonist BRL49653 to potentiate lipid body formation and PGE(2) production; furthermore, pretreatment with the PPARgamma antagonist GW9662 inhibited BCG-induced lipid body formation and PGE(2) production. BCG-induced MIP 1alpha, IL12p70, TNF-alpha, and IL6 production was not inhibited by GW9662 treatment. Nonpathogenic Mycobacterium smegmatis failed to induce PPARgamma expression or lipid body formation. Moreover, inhibition of PPARgamma by GW9662 enhanced the mycobacterial killing capacity of macrophages. Our findings show that PPARgamma is involved in lipid body biogenesis, unravels a cross-talk between the innate immune receptor TLR2 and the lipid-activated nuclear receptor PPARgamma that coordinates lipid metabolism and inflammation in BCG-infected macrophages, thereby potentially affecting mycobacterial pathogenesis. PMID- 19561093 TI - Immunomodulatory function of bone marrow-derived mesenchymal stem cells in experimental autoimmune type 1 diabetes. AB - Human clinical trials in type 1 diabetes (T1D) patients using mesenchymal stem cells (MSC) are presently underway without prior validation in a mouse model for the disease. In response to this void, we characterized bone marrow-derived murine MSC for their ability to modulate immune responses in the context of T1D, as represented in NOD mice. In comparison to NOD mice, BALB/c-MSC mice were found to express higher levels of the negative costimulatory molecule PD-L1 and to promote a shift toward Th2-like responses in treated NOD mice. In addition, transfer of MSC from resistant strains (i.e., nonobese resistant mice or BALB/c), but not from NOD mice, delayed the onset of diabetes when administered to prediabetic NOD mice. The number of BALB/c-MSC trafficking to the pancreatic lymph nodes of NOD mice was higher than in NOD mice provided autologous NOD-MSC. Administration of BALB/c-MSC temporarily resulted in reversal of hyperglycemia in 90% of NOD mice (p = 0.002). Transfer of autologous NOD-MSC imparted no such therapeutic benefit. We also noted soft tissue and visceral tumors in NOD-MSC treated mice, which were uniquely observed in this setting (i.e., no tumors were present with BALB/c- or nonobese resistant mice-MSC transfer). The importance of this observation remains to be explored in humans, as inbred mice such as NOD may be more susceptible to tumor formation. These data provide important preclinical data supporting the basis for further development of allogeneic MSC-based therapies for T1D and, potentially, for other autoimmune disorders. PMID- 19561095 TI - TLR9 activation coupled to IL-10 deficiency induces adverse pregnancy outcomes. AB - Pregnancy outcome is severely compromised by intrauterine infections and inflammation. Although the pregnant uterine microenvironment is replete with innate immune cells and TLR expression, the mechanisms that facilitate adverse effects of their activation are largely unknown. In this study, we mimic the activation of TLR9 with its pathogenic ligand hypomethylated CpG and demonstrate that IL-10 proficiency protects against CpG-induced pregnancy complications. We show that fetal resorption and preterm birth are rapidly induced in IL-10(-/-) mice by low doses of CpG (approximately 25 microg/mouse) when injected i.p. on gestational day 6 or gestational day 14, respectively. In contrast, wild-type mice failed to experience such effects at comparable doses, but pups born at term displayed craniofacial/limb defects in response to higher doses (approximately 400 microg/mouse). Pregnancy complications in IL-10(-/-) mice were associated with unexpected and robust TLR9-triggered activation and amplification of uterine neutrophil and macrophage subpopulations followed by their migration to the placental zone. Furthermore, a dramatic increase in serum levels of mouse KC and TNF-alpha production by uterine F4/80(+) cells, but not uterine NK or Gr 1(+)CD11b(+) cells, was observed. Depletion of F4/80(+) macrophages or neutralization of TNF-alpha rescued pregnancy to term. Our results have important implications for IL-10-mediated "uterine tolerance" against CpG-driven innate immune activation. PMID- 19561096 TI - Role of MAPK kinase 6 in arthritis: distinct mechanism of action in inflammation and cytokine expression. AB - Development of p38alpha inhibitors for rheumatoid arthritis has been hindered by toxicity and limited efficacy. Therefore, we evaluated whether MKK6, an upstream kinase that regulates multiple p38 isoforms, might be an alternative therapeutic target in inflammatory arthritis. Wild-type (WT), MKK6(-/-), and MKK3(-/-) mice were administered K/BxN serum to induce arthritis. Articular expression of activated kinases and cytokines was determined by Western blot, qPCR, ELISA, and multiplex analysis. Immunoprecipitation and confocal microscopy experiments were performed to determine the subcellular location of MKK6, P-p38, and MAPKAPK2 (MK2). Arthritis scores were significantly lower in MKK6(-/-) mice compared with WT mice. Joint destruction and osteoclast differentiation were lower in MKK6(-/ ), as were articular IL-6 and matrix metalloproteinase-3 expression. Phospho-p38 levels were modestly decreased in the joints of arthritic MKK6(-/-) mice compared with WT but were significantly higher than MKK3(-/-) mice. P-MK2 was low in MKK6( /-) and MKK3(-/-) mice. Uncoupled p38 and MK2 activation was also observed in cultured, MKK6(-/-) FLS and confirmed using kinase assays. Immunoprecipitation assays and confocal microscopy showed that P-p38 and MK2 colocalized in activated WT but not MKK6(-/-) FLS. Distinct patterns of cytokine production were observed in MKK6(-/-) and MKK3(-/-) cells. MKK6 deficiency suppresses inflammatory arthritis and joint destruction, suggesting it might be a therapeutic target for inflammation. Although MKK3 and MKK6 activate the p38 pathway, they regulate distinct subsets of proinflammatory cytokines. MKK6 appears mainly to facilitate p38 and MK2 colocalization in the nucleus rather than to phosphorylate p38. PMID- 19561097 TI - Lethal effect of CD3-specific antibody in mice deficient in TGF-beta1 by uncontrolled flu-like syndrome. AB - CD3-specific Ab therapy results in a transient, self-limiting, cytokine associated, flu-like syndrome in experimental animals and in patients, but the underlying mechanism for this spontaneous resolution remains elusive. By using an in vivo model of CD3-specific Ab-induced flu-like syndrome, we show in this paper that a single injection of sublethal dose of the Ab killed all TGF-beta1(-/-) mice. The death of TGF-beta1(-/-) mice was associated with occurrence of this uncontrolled flu-like syndrome, as demonstrated by a sustained storm of systemic inflammatory TNF and IFN-gamma cytokines. We present evidence that deficiency of professional phagocytes to produce TGF-beta1 after apoptotic T cell clearance may be responsible, together with hypersensitivity of T cells to both activation and apoptosis, for the uncontrolled inflammation. These findings indicate a key role for TGF-beta1 and phagocytes in protecting the recipients from lethal inflammation and resolving the flu-like syndrome after CD3-specific Ab treatment. The study may also provide a novel molecular mechanism explaining the early death in TGF-beta1(-/-) mice. PMID- 19561098 TI - Treatment with a C5aR antagonist decreases pathology and enhances behavioral performance in murine models of Alzheimer's disease. AB - Alzheimer's disease (AD) is an age-related dementia, characterized by amyloid plaques, neurofibrillary tangles, neuroinflammation, and neuronal loss in the brain. Components of the complement system, known to produce a local inflammatory reaction, are associated with the plaques and tangles in AD brain, and thus a role for complement-mediated inflammation in the acceleration or progression of disease has been proposed. A complement activation product, C5a, is known to recruit and activate microglia and astrocytes in vitro by activation of a G protein-coupled cell-surface C5aR. Here, oral delivery of a cyclic hexapeptide C5a receptor antagonist (PMX205) for 2-3 mo resulted in substantial reduction of pathological markers such as fibrillar amyloid deposits (49-62%) and activated glia (42-68%) in two mouse models of AD. The reduction in pathology was correlated with improvements in a passive avoidance behavioral task in Tg2576 mice. In 3xTg mice, PMX205 also significantly reduced hyperphosphorylated tau (69%). These data provide the first evidence that inhibition of a proinflammatory receptor-mediated function of the complement cascade (i.e., C5aR) can interfere with neuroinflammation and neurodegeneration in AD rodent models, suggesting a novel therapeutic target for reducing pathology and improving cognitive function in human AD patients. PMID- 19561099 TI - Pulmonary inflammation triggered by ricin toxin requires macrophages and IL-1 signaling. AB - Ricin is a potent ribotoxin considered to be a potentially dangerous bioterrorist agent due to its wide availability and the possibility of aerosol delivery to human populations. Studies in rodents and nonhuman primates have demonstrated that ricin delivered to the pulmonary system leads to acute lung injury and symptoms resembling acute respiratory distress syndrome. Increasing evidence suggests that the inflammatory effects triggered by ricin are responsible for its lethality. We demonstrated previously that ricin administered to the lungs of mice causes death of pulmonary macrophages and the release of proinflammatory cytokines, suggesting macrophages may be a primary target of ricin. Here we examined the requirement for macrophages in the development of ricin-mediated pulmonary inflammation by employing transgenic (MAFIA) mice that express an inducible gene driven by the c-fms promoter for Fas-mediated apoptosis of macrophages upon injection of a synthetic dimerizer, AP20187. Administration of aerosolized ricin to macrophage-depleted mice led to reduced inflammatory responses, including recruitment of neutrophils, expression of proinflammatory transcripts, and microvascular permeability. When compared with control mice treated with ricin, macrophage-depleted mice treated with ricin displayed a reduction in pulmonary IL-1beta. Employing mice deficient in IL-1, we found that ricin-induced inflammatory responses were suppressed, including neutrophilia. Neutrophilia could be restored by co-administering ricin and exogenous IL-1beta to IL-1alpha/beta(-/-) mice. Furthermore, IL1Ra/anakinra cotreatment inhibited ricin-mediated inflammatory responses, including recruitment of neutrophils, expression of proinflammatory genes, and histopathology. These data suggest a central role for macrophages and IL-1 signaling in the inflammatory process triggered by ricin. PMID- 19561101 TI - An activating and inhibitory signal from an inhibitory receptor LMIR3/CLM-1: LMIR3 augments lipopolysaccharide response through association with FcRgamma in mast cells. AB - Leukocyte mono-Ig-like receptor 3 (LMIR3) is an inhibitory receptor mainly expressed in myeloid cells. Coengagement of Fc epsilonRI and LMIR3 impaired cytokine production in bone marrow-derived mast cells (BMMCs) induced by Fc epsilonRI crosslinking alone. Mouse LMIR3 possesses five cytoplasmic tyrosine residues (Y241, Y276, Y289, Y303, Y325), among which Y241 and Y289 (Y241/289) or Y325 fit the consensus sequence of ITIM or immunotyrosine-based switch motif (ITSM), respectively. The inhibitory effect was abolished by the replacement of Y325 in addition to Y241/289 with phenylalanine (Y241/189/325/F) in accordance with the potential of Y241/289/325 to cooperatively recruit Src homology region 2 domain-containing phosphatase 1 (SHP)-1 or SHP-2. Intriguingly, LMIR3 crosslinking alone induced cytokine production in BMMCs expressing LMIR3 (Y241/276/289/303/325F) mutant as well as LMIR3 (Y241/289/325F). Moreover, coimmunoprecipitation experiments revealed that LMIR3 associated with ITAM containing FcRgamma. Analysis of FcRgamma-deficient BMMCs demonstrated that both Y276/303 and FcRgamma played a critical role in the activating function of this inhibitory receptor. Importantly, LMIR3 crosslinking enhanced cytokine production of BMMCs stimulated by LPS, while suppressing production stimulated by other TLR agonists or stem cell factor. Thus, an inhibitory receptor LMIR3 has a unique property to associate with FcRgamma and thereby functions as an activating receptor in concert with TLR4 stimulation. PMID- 19561100 TI - Autocrine IL-10 induces hallmarks of alternative activation in macrophages and suppresses antituberculosis effector mechanisms without compromising T cell immunity. AB - Elevated IL-10 has been implicated in reactivation tuberculosis (TB). Since macrophages rather than T cells were reported to be the major source of IL-10 in TB, we analyzed the consequences of a macrophage-specific overexpression of IL-10 in transgenic mice (macIL-10-transgenic) after aerosol infection with Mycobacterium tuberculosis (Mtb). MacIL-10 transgenic mice were more susceptible to chronic Mtb infection than nontransgenic littermates, exhibiting higher bacterial loads in the lung after 12 wk of infection and dying significantly earlier than controls. The differentiation, recruitment, and activation of Th1 cells as well as the induction of IFN-gamma-dependent effector genes against Mtb were not affected by macrophage-derived IL-10. However, microarray analysis of pulmonary gene expression revealed patterns characteristic of alternative macrophage activation that were overrepresented in Mtb-infected macIL-10 transgenic mice. Importantly, arginase-1 gene expression and activity were strikingly enhanced in transgenic mice accompanied by a reduced production of reactive nitrogen intermediates. Moreover, IL-10-dependent arginase-1 induction diminished antimycobacterial effector mechanisms in macrophages. Taken together, macrophage-derived IL-10 triggers aspects of alternative macrophage activation and promotes Mtb recrudescence independent of overt effects on anti-TB T cell immunity. PMID- 19561102 TI - Heat shock protein 60, via MyD88 innate signaling, protects B cells from apoptosis, spontaneous and induced. AB - We recently reported that heat shock protein 60 (HSP60) via TLR4 signaling activates B cells and induces them to proliferate and secrete IL-10. We now report that HSP60 inhibits mouse B cell apoptosis, spontaneous or induced by dexamethasone or anti-IgM activation. Unlike HSP60 enhancement of B cell proliferation and IL-10 secretion, TLR4 signaling was not required for the inhibition of apoptosis by HSP60; nevertheless, MyD88 was essential. Inhibition of apoptosis by HSP60 was associated with up-regulation of the antiapoptotic molecules Bcl-2, Bcl-x(L), and survivin, maintenance of the mitochondrial transmembrane potential, and inhibition of caspase-3 activation. Moreover, B cells incubated with HSP60 manifested prolonged survival following transfer into recipient mice. These results extend the varied role of HSP60 in the innate regulation of the adaptive immune response. PMID- 19561103 TI - Activation of the Akt-NF-kappaB pathway by subtilase cytotoxin through the ATF6 branch of the unfolded protein response. AB - Shiga toxin has the potential to induce expression of inflammation-associated genes, although the underlying mechanisms are not well understood. We examined the effects of subtilase cytotoxin (SubAB), an AB(5) toxin produced by some Shiga toxigenic Escherichia coli, on the activation of NF-kappaB. SubAB is known to be a protease which selectively degrades GRP78/Bip. Treatment of NRK-52E cells with SubAB caused rapid cleavage of GRP78. Following the degradation of GRP78, transient activation of NF-kappaB was observed with a peak at 6-12 h; the activation subsided within 24 h despite the continuous absence of intact GRP78. The activation of NF-kappaB was preceded by transient phosphorylation of Akt. Treatment of the cells with a selective inhibitor of Akt1/2 or an inhibitor of PI3K attenuated SubAB-induced NF-kappaB activation, suggesting that activation of Akt is an event upstream of NF-kappaB. Degradation of GRP78 caused the unfolded protein response (UPR), and inducers of the UPR mimicked the stimulatory effects of SubAB on Akt and NF-kappaB. SubAB triggered the three major branches of the UPR including the IRE1-XBP1, PERK, and ATF6 pathways. Dominant-negative inhibition of IRE1alpha, XBP1, or PERK did not attenuate activation of NF-kappaB by SubAB. In contrast, genetic and pharmacological inhibition of ATF6 significantly suppressed SubAB-triggered Akt phosphorylation and NF-kappaB activation. These results suggested that loss of GRP78 by SubAB leads to transient phosphorylation of Akt and consequent activation of NF-kappaB through the ATF6 branch of the UPR. PMID- 19561104 TI - The opposite effects of acute and chronic alcohol on lipopolysaccharide-induced inflammation are linked to IRAK-M in human monocytes. AB - Impaired host defense after alcohol use is linked to altered cytokine production, however, acute and chronic alcohol differently modulate monocyte/macrophage activation. We hypothesized that in human monocytes, acute alcohol induces hyporesponsiveness to LPS, resulting in decreased TNF-alpha, whereas chronic alcohol increases TNF-alpha by sensitization to LPS. We found that acute alcohol increased IL-1R-associated kinase-monocyte (IRAK-M), a negative regulator of IRAK 1, in human monocytes. This was associated with decreased IkappaB alpha kinase activity, NFkappaB DNA binding, and NFkappaB-driven reporter activity after LPS stimulation. In contrast, chronic alcohol decreased IRAK-M expression but increased IRAK-1 and IKK kinase activities, NFkappaB DNA binding, and NFkappaB reporter activity. Inhibition of IRAK-M in acute alcohol-exposed monocytes using small interfering RNA restored the LPS-induced TNF-alpha production whereas over expression of IRAK-M in chronic alcohol macrophages prevented the increase in TNF alpha production. Addition of inhibitors of alcohol metabolism did not alter LPS signaling and TNF-alpha production during chronic alcohol exposure. IRAK-1 activation induces MAPKs that play an important role in TNF-alpha induction. We determined that acute alcohol decreased but chronic alcohol increased activation of ERK in monocytes and ERK inhibitor, PD98059, prevented the chronic alcohol induced increase in TNF-alpha. In summary, inhibition of LPS-induced NFkappaB and ERK activation by acute alcohol leads to hyporesponsiveness of monocytes to LPS due to increased IRAK-M. In contrast, chronic alcohol sensitizes monocytes to LPS through decreased IRAK-M expression and activation of NFkappaB and ERK kinases. Our data indicate that IRAK-M is a central player in the opposite regulation of LPS signaling by different lengths of alcohol exposure in monocytes. PMID- 19561105 TI - Patent filarial infection modulates malaria-specific type 1 cytokine responses in an IL-10-dependent manner in a filaria/malaria-coinfected population. AB - The effect of filarial infections on malaria-specific immune responses was investigated in Malian villages coendemic for filariasis (Fil) and malaria. Cytokines were measured from plasma and Ag-stimulated whole blood from individuals with Wuchereria bancrofti and/or Mansonella perstans infections (Fil(+); n = 19) and those without evidence of filarial infection (Fil(-); n = 19). Plasma levels of IL-10 (geometric mean [GM], 22.8 vs 10.4) were higher in Fil(+) compared with Fil(-), whereas levels of IFN-inducible protein (IP)-10 were lower in Fil(+) (GM, 66.3 vs 110.0). Fil(+) had higher levels of spontaneously secreted IL-10 (GM, 59.3 vs 6.8 pg/ml) and lower levels of IL-2 (1.0 vs 1.2 pg/ml) than did Fil(-). Although there were no differences in levels of Staphylococcus aureus enterotoxin B-induced cytokines between the two groups, Fil(+) mounted lower IL-12p70 (GM, 1.11 vs 3.83 pg/ml; p = 0.007), IFN-gamma (GM, 5.44 vs 23.41 pg/ml; p = 0.009), and IP-10 (GM, 29.43 vs 281.7 pg/ml; p = 0.007) responses following malaria Ag (MalAg) stimulation compared with Fil(-). In contrast, Fil(+) individuals had a higher MalAg-specific IL-10 response (GM, 7318 pg/ml vs 3029 pg/ml; p = 0.006) compared with those without filarial infection. Neutralizing Ab to IL-10 (but not to TGFbeta) reversed the down-regulated MalAg specific IFN-gamma and IP-10 (p < 0.001) responses in Fil(+). Together, these data demonstrate that filarial infections modulate the Plasmodium falciparum specific IL-12p70/IFN-gamma secretion pathways known to play a key role in resistance to malaria and that they do so in an IL-10-dependent manner. PMID- 19561107 TI - Pathogen-induced interleukin-1beta processing and secretion is regulated by a biphasic redox response. AB - In this study, we show that IL-1beta processing and secretion induced by pathogen associated molecular pattern (PAMP) molecules in human monocytes is regulated by a biphasic redox event including a prompt oxidative stress and a delayed antioxidant response. Namely, PAMPs induce an early generation of reactive oxygen species (ROS) followed by increase of intracellular thioredoxin and release of reduced cysteine: this antioxidant phase is paralleled by secretion of mature IL 1beta. ROS production and antioxidant response are both required, because either inhibitors of NADPH oxidase and of thioredoxin reductase impair IL-1beta secretion. These inhibitors also hinder cysteine release and consequently prevent reduction of the extracellular medium: addition of exogenous reducing agents restores IL-1beta secretion. Not only silencing of thioredoxin, but also of the ROS scavenger superoxide dismutase 1 results in inhibition of IL-1beta secretion. Thus, PAMP-induced ROS trigger an antioxidant response involving intracellular redox enzymes and release of cysteine, ultimately required for IL-1beta processing and secretion. PMID- 19561106 TI - T cell responses in the absence of IFN-gamma exacerbate uterine infection with Chlamydia trachomatis. AB - Infection with the obligate intracellular bacterium Chlamydia trachomatis is controlled primarily by IFN-gamma and Th1 immunity. In this study, we used cells from a Chlamydia-specific CD4(+) TCR-transgenic mouse to assess the role of IFN gamma in development of Th1 immunity. We show that secretion of host IFN-gamma or the ability of host cells to respond to secreted IFN-gamma is not required to initiate a Th1 immune response. Additionally, we found that Ag-specific CD4(+) cells that were preskewed toward Th1 confer protection, whereas cells preskewed toward Th2 cause a previously unreported exacerbation of disease leading to higher bacterial load. Chlamydia-specific Th1 cells transferred into an IFN gamma(-/-) recipient mouse demonstrate protective effects, but the same cells exacerbate bacterial burden when transferred into IFN-gammaR(-/-) mice. Thus, we demonstrate that the secretion of IFN-gamma is necessary for protection against C. trachomatis and that in the absence of host cell IFN-gammaR expression, both Th1 and Th2 cells lead to increased burden of C. trachomatis. PMID- 19561108 TI - A matter of timing: unsynchronized antigen expression and antigen presentation diminish secondary T cell responses. AB - Despite the low and short expression of secondary Ag, prime-boost immunizations using homologous or heterologous vectors are capable of amplifying memory CD8(+) T cells. This is mainly attributed to the rapid presentation of Ag by APCs and the high proliferative capacity of memory CD8(+) T cells. Nevertheless, certain viruses and vectors often require prolonged Ag presentation for optimal T cell priming, and the influence of such a prolonged presentation during secondary immune induction is not clear. To address this issue, we primed and boosted mice intradermally (i.d.) with plasmid DNA that was recently reported to require prolonged Ag presentation for maximal CD8(+) T cell priming. Although functional memory CD8(+) T cells were present in the mice after i.d. priming, the secondary CD8(+) T cell response elicited was limited and reached a similar level of that observed during priming. The initial levels of secondary Ag expressed in the boosted mice were sufficient to prime CD8(+) T cell response in naive hosts, suggesting that lower Ag load alone does not explain the limited secondary immune responses observed. Removal of the injection site 5 or 10 days after i.d. boosting immunization resulted in diminished Ag presentation and no expansion of memory CD8(+) T cells. In fact, Ag-presenting activity following boost occurred mainly two weeks postimmunization, a time when the Ag was no longer expressed in situ. These findings suggest that when the boosting vector triggers prolonged Ag presentation, the lack of synchronicity between Ag accessibility and Ag presentation limits secondary immune responses. PMID- 19561109 TI - Proteases induce production of thymic stromal lymphopoietin by airway epithelial cells through protease-activated receptor-2. AB - Thymic stromal lymphopoietin (TSLP) is produced by epithelial cells and triggers dendritic cell-mediated Th2-type inflammation. Although TSLP is up-regulated in epithelium of patients with asthma, the factors that control TSLP production have not been studied extensively. Because mouse models suggest roles for protease(s) in Th2-type immune responses, we hypothesized that proteases from airborne allergens may induce TSLP production in a human airway epithelial cell line, BEAS 2B. TSLP mRNA and protein were induced when BEAS-2B cells were exposed to prototypic proteases, namely, trypsin and papain. TSLP induction by trypsin required intact protease activity and also a protease-sensing G protein-coupled receptor, protease-activated receptor (PAR)-2; TSLP induction by papain was partially dependent on PAR-2. In humans, exposure to ubiquitous airborne fungi, such as Alternaria, is implicated in the development and exacerbation of asthma. When BEAS-2B cells or normal human bronchial epithelial cells were exposed to Alternaria extract, TSLP was potently induced. The TSLP-inducing activity of Alternaria was partially blocked by treating the extract with a cysteine protease inhibitor, E-64, or by infecting BEAS-2B cells with small interfering RNA for PAR 2. Protease-induced TSLP production by BEAS-2B cells was enhanced synergistically by IL-4 and abolished by IFN-gamma. These findings demonstrate that TSLP expression is induced in airway epithelial cells by exposure to allergen-derived proteases and that PAR-2 is involved in the process. By promoting TSLP production in the airways, proteases associated with airborne allergens may facilitate the development and/or exacerbation of Th2-type airway inflammation, particularly in allergic individuals. PMID- 19561110 TI - The role for T cell repertoire/antigen-specific interactions in experimental kidney ischemia reperfusion injury. AB - T cells have been implicated in the early pathogenesis of ischemia reperfusion injury (IRI) of kidney, liver, lung, and brain. It is not known whether Ag-TCR engagement followed by Ag-specific T cell activation participates in IRI. T cell deficient nu/nu mice are moderately resistant to renal IRI, which can be reversed upon reconstitution with syngeneic T cells. In this study, we found that nu/nu mice reconstituted with DO11.10 T cells, limited in their TCR repertoire, have significantly less kidney dysfunction and tubular injury after renal IRI compared with that in nu/nu mice reconstituted with wild-type T cells having a diverse TCR repertoire. CD4(+) T cells infiltrating ischemic kidneys of nu/nu mice reconstituted with DO11.10 T cells exhibited lower IFN-gamma production than that of wild-type controls. Frequency of regulatory T cells in kidneys of these mice was similar in both DO11.10 T cells and wild-type T cell recipient groups. DO11.10 mice immunized with OVA-CFA had significantly worse kidney function at 24 h after ischemia than those immunized with CFA alone. Thus, without T cell activation, diverse TCR repertoire was important for renal IRI in naive mice. However, once T cells were activated in an Ag-specific manner through TCR in DO11.10 mice, a restricted TCR repertoire no longer limited the extent of kidney injury. Thus, both TCR repertoire-dependent and -independent factors mediate T cell functions in kidney IRI. PMID- 19561111 TI - Targeted liposomal delivery of TLR9 ligands activates spontaneous antitumor immunity in an autochthonous cancer model. AB - Accessibility of tumors for highly effective local treatment represents a major challenge for anticancer therapy. Immunostimulatory oligodeoxynucleotides (ODN) with CpG motifs are ligands of TLR9, which prime spontaneous antitumor immunity, but are less effective when applied systemically. We therefore developed a liposome-based agent for selective delivery of CpG-ODN into the tumor environment. A peptide that specifically targets angiogenic endothelial cells in a transgenic tumor model for islet cell carcinogenesis was engrafted into CpG-ODN containing liposomes. Intravenous injection of these liposomes resulted in specific accumulation around tumor vessels, increased uptake by tumor-resident macrophages, and retention over time. In contrast, nontargeted liposomes did not localize to the tumor vasculature. Consequently, only vascular targeting of CpG ODN liposomes provoked a marked inflammatory response at vessel walls with enhanced CD8(+) and CD4(+) T cell infiltration and, importantly, activation of spontaneous, tumor-specific cytotoxicity. In a therapeutic setting, 40% of tumor bearing, transgenic mice survived beyond week 45 after systemic administration of vascular-directed CpG-ODN liposomes. In contrast, control mice survived up to 30 wk. Therapeutic efficacy was further improved by increasing the frequency of tumor-specific effector cells through adoptive transfers. NK cells and CD8(+) T cells were major effectors which induced tumor cell death and acted in conjunction with antivascular effects. Thus, tumor homing with CpG-ODN-loaded liposomes is as potent as direct injection of free CpG-ODN and has the potential to overcome some major limitations of conventional CpG-ODN monotherapy. PMID- 19561113 TI - Mechanisms of opioid-mediated inhibition of human T cell receptor signaling. AB - Opioids are widely used for the treatment of severe pain. However, it is also known that opioids, in particular morphine, cause immunosuppression. Therefore, their use may complicate treatment of persons with an already impaired immune system, e.g., patients suffering from cancer or AIDS. We investigated the mechanisms of opioid-induced immunosuppression in primary human T lymphocytes and the human T cell line Jurkat. We demonstrated that morphine and the endogenous opioid beta-endorphin inhibited the transcription of IL-2 in activated human T lymphocytes as well as the activation of the transcription factors AP-1, NFAT, and NF-kappaB, which transactivate IL-2. In addition, the TCR-induced calcium flux and MAPK activation were inhibited by the opioids, as well as proximal signaling events, such as the phosphorylation of the linker for activation of T cells and Zap70. A more detailed characterization of the mechanism revealed that incubation of T cells with the opioids caused a marked increase in cAMP. This in turn activated protein kinase A, which augmented the kinase activity of C terminal Src kinase bound to phosphoprotein associated with glycosphingolipid enrich microdomains, resulting in a further enhancement of the tonic inhibition of the leukocyte-specific protein tyrosine kinase Lck, thereby blocking the initiation of TCR signaling. These effects were mediated by mu opioid receptors. Our findings contribute to the understanding of immunosuppressive side effects of morphine. Since beta-endorphin is expressed and secreted by immune effector cells, including T cells, and up-regulated in these cells by various stimuli, our data also suggest an inhibitory role for beta-endorphin in the physiological regulation of T cell activation. PMID- 19561114 TI - Incomplete Carney triad--a review of two cases. AB - A curious association of three rare tumours was described by Carney in 1977. 'Carney's triad' characteristically includes multifocal pulmonary chondroma, gastric stromal sarcoma and extra-adrenal paraganglioma. Patients may exhibit complete or incomplete expression of the triad. Carney acknowledged that, of 79 patients, only 17 possessed all three tumours. We report here two patients with incomplete expression of Carney's triad. PMID- 19561112 TI - Focal adhesion kinase regulates pathogen-killing capability and life span of neutrophils via mediating both adhesion-dependent and -independent cellular signals. AB - Various neutrophil functions such as phagocytosis, superoxide production, and survival are regulated by integrin signaling. Despite the essential role of focal adhesion kinase (FAK) in mediating this signaling pathway, its exact function in neutrophils is ill defined. In this study, we investigated the role of FAK in neutrophils using a myeloid-specific conditional FAK knockout mouse. As reported in many other cell types, FAK is required for regulation of focal adhesion dynamics when neutrophils adhere to fibronectin or ICAM-1. Adhesion on VCAM-1 coated surfaces and chemotaxis after adhesion were not altered in FAK null neutrophils. In addition, we observed significant reduction in NADPH oxidase mediated superoxide production and complement-mediated phagocytosis in FAK null neutrophils. As a result, these neutrophils displayed decreased pathogen killing capability both in vitro and in vivo in a mouse peritonitis model. In adherent cells, the defects associated with FAK deficiency are likely due to suppression of phosphatidylinositol (3,4,5)-trisphosphate (PtdIns(3,4,5)P3) signaling and chemoattractant-elicited calcium signaling. Disruption of FAK also reduced chemoattractant-elicited superoxide production in suspended neutrophils in the absence of cell adhesion. This may be solely caused by suppression of PtdIns(3,4,5)P3 signaling in these cells, because the fMLP-elicited calcium signal was not altered. Consistent with decreased PtdIns(3,4,5)P3/Akt signaling in FAK null neutrophils, we also observed accelerated spontaneous death in these cells. Taken together, our results revealed previously unrecognized roles of FAK in neutrophil function and provided a potential therapeutic target for treatment of a variety of infectious and inflammatory diseases. PMID- 19561116 TI - Pedunculated polyp of early sigmoid colon cancer with invasive micropapillary carcinoma. AB - A 64-year-old man was admitted to Dongo Hospital (Nara, Japan) with colonic cancer, following the onset of abdominal pain, diarrhea and fever. A pedunculated polyp was detected in the sigmoid colon by colonoscopy, and laparoscopy-assisted sigmoidectomy with regional lymph node resection was performed. Histopathologically, the tumor exhibited massive invasion of the submucosa, and multiple lymph node metastases were detected. The tumor mainly consisted of a micropapillary component. Immunohistochemically, MUC1 was expressed at the stromal edge of the micropapillary component and showed the characteristic 'inside-out' pattern of a micropapillary carcinoma. The multiple lymph node metastases were predominantly composed of carcinoma with a micropapillary pattern. Our case suggests that when a micropapillary component is identified in a pre-operative biopsy specimen, even for a pedunculated early colorectal cancer, the extent of surgical resection should be carefully considered due to the high potential for nodal metastasis. PMID- 19561115 TI - QJM. Elements: in this month's issue. PMID- 19561117 TI - High blood pressure in Latin America: a call to action. AB - High blood pressure (BP) is an enormous global problem, and is especially challenging for low- and middle-income countries such as those of Latin America. Although developed countries have benefited from significant reductions in cardiovascular and cerebrovascular disease in recent decades, comparable reductions have not been achieved in Latin America. In fact, the prevalence of high BP is increasing in many Latin American countries, and the situation will worsen without definitive efforts to correct it. The growing preponderance of hypertension and chronic diseases, coupled with expected increases in population growth, present a mounting threat to Latin American economies. This report provides a comprehensive overview of the burden of high BP throughout Latin America, and presents recommendations for change. The dismal observations warrant a call to action for improved control of high BP and other cardiovascular risk factors across Latin America. Achieving these ambitious goals will require collaborative efforts by many groups, including policymakers, international organizations, healthcare providers, schools and society as a whole. PMID- 19561118 TI - The cap binding complex influences H2B ubiquitination by facilitating splicing of the SUS1 pre-mRNA. AB - Pre-messenger RNA splicing is carried out by a large ribonucleoprotein complex called the spliceosome. Despite the striking evolutionary conservation of the spliceosomal components and their functions, controversy persists about the relative importance of splicing in Saccharomyces cerevisiae-particularly given the paucity of intron-containing genes in yeast. Here we show that splicing of one pre-messenger RNA, SUS1, a component of the histone H2B ubiquitin protease machinery, is essential for establishing the proper modification state of chromatin. One protein complex that is intimately involved in pre-mRNA splicing, the yeast cap-binding complex, appears to be particularly important, as evidenced by its extensive and unique genetic interactions with enzymes that catalyze histone H2B ubiquitination. Microarray studies show that cap binding complex (CBC) deletion has a global effect on gene expression, and for approximately 20% of these genes, this effect is suppressed when ubiquitination of histone H2B is eliminated. Consistent with this finding of histone H2B dependent effects on gene expression, deletion of the yeast cap binding complex leads to overubiquitination of histone H2B. A key component of the ubiquitin-protease module of the SAGA complex, Sus1, is encoded by a gene that contains two introns and is misspliced when the CBC is deleted, leading to destabilization of the ubiquitin protease complex and defective modulation of cellular H2B levels. These data demonstrate that pre-mRNA splicing plays a critical role in histone H2B ubiquitination and that the CBC in particular helps to establish the proper state of chromatin and proper expression of genes that are regulated at the level of histone H2B ubiquitination. PMID- 19561121 TI - Turnover of fatty acids during natural senescence of Arabidopsis, Brachypodium, and switchgrass and in Arabidopsis beta-oxidation mutants. AB - During leaf senescence, macromolecule breakdown occurs and nutrients are translocated to support growth of new vegetative tissues, seeds, or other storage organs. In this study, we determined the fatty acid levels and profiles in Arabidopsis (Arabidopsis thaliana), Brachypodium distachyon, and switchgrass (Panicum virgatum) leaves during natural senescence. In young leaves, fatty acids represent 4% to 5% of dry weight and approximately 10% of the chemical energy content of the leaf tissues. In all three species, fatty acid levels in leaves began to decline at the onset of leaf senescence and progressively decreased as senescence advanced, resulting in a greater than 80% decline in fatty acids on a dry weight basis. During senescence, Arabidopsis leaves lost 1.6% of fatty acids per day at a rate of 2.1 mug per leaf (0.6 mug mg(-1) dry weight). Triacylglycerol levels remained less than 1% of total lipids at all stages. In contrast to glycerolipids, aliphatic surface waxes of Arabidopsis leaves were much more stable, showing only minor reduction during senescence. We also examined three Arabidopsis mutants, acx1acx2, lacs6lacs7, and kat2, which are blocked in enzyme activities of beta-oxidation and are defective in lipid mobilization during seed germination. In each case, no major differences in the fatty acid contents of leaves were observed between these mutants and the wild type, indicating that several mutations in beta-oxidation that cause reduced breakdown of reserve oil in seeds do not substantially reduce the degradation of fatty acids during leaf senescence. PMID- 19561120 TI - The mechanism of iron homeostasis in the unicellular cyanobacterium synechocystis sp. PCC 6803 and its relationship to oxidative stress. AB - In this article, we demonstrate the connection between intracellular iron storage and oxidative stress response in cyanobacteria. Iron is essential for the survival of all organisms. However, the redox properties that make iron a valuable cofactor also lead to oxidative interactions, resulting in the formation of harmful radicals. Therefore, iron accumulation in cells should be tightly regulated, a process in which ferritin family proteins play an important role. Synechocystis sp. PCC 6803 contains two ferritin-type storage complexes, bacterioferritin and MrgA. Previous studies demonstrated the role of bacterioferritin and MrgA in iron storage. In addition, MrgA was found to play a key role in oxidative stress response. Here, we examined the dual role of the ferritin family proteins using physiological and transcriptomic approaches. Microarray analysis of iron-limited wild-type and DeltamrgA cultures revealed a substantial up-regulation of oxidative stress-related genes in mutant cells. The PerR regulator was found to play an important role in that process. Furthermore, we were able to demonstrate the connection between internal iron quota, the presence of the two storage complexes, and the sensitivity to externally applied oxidative stress. These data suggest a pivotal role for the ferritin-type proteins of Synechocystis sp. PCC 6803 in coordinating iron homeostasis and in oxidative stress response. The combined action of the two complexes allows for the safe accumulation and release of iron from storage by minimizing damage resulting from interactions between reduced iron and the oxygen radicals that are produced in abundance by the photosynthetic apparatus. PMID- 19561122 TI - Underexplored niches in research on plant pathogenic bacteria. PMID- 19561119 TI - Emerging roles of microRNAs as molecular switches in the integrated circuit of the cancer cell. AB - Transformation of normal cells into malignant tumors requires the acquisition of six hallmark traits, e.g., self-sufficiency in growth signals, insensitivity to antigrowth signals and self-renewal, evasion of apoptosis, limitless replication potential, angiogenesis, invasion, and metastasis, which are common to all cancers (Hanahan and Weinberg 2000). These new cellular traits evolve from defects in major regulatory microcircuits that are fundamental for normal homeostasis. The discovery of microRNAs (miRNAs) as a new class of small non protein-coding RNAs that control gene expression post-transcriptionally by binding to various mRNA targets suggests that these tiny RNA molecules likely act as molecular switches in the extensive regulatory web that involves thousands of transcripts. Most importantly, accumulating evidence suggests that numerous microRNAs are aberrantly expressed in human cancers. In this review, we discuss the emergent roles of microRNAs as switches that function to turn on/off known cellular microcircuits. We outline recent compelling evidence that deregulated microRNA-mediated control of cellular microcircuits cooperates with other well established regulatory mechanisms to confer the hallmark traits of the cancer cell. Furthermore, these exciting insights into aberrant microRNA control in cancer-associated circuits may be exploited for cancer therapies that will target deregulated miRNA switches. PMID- 19561124 TI - A novel domain in histone deacetylase 1 and 2 mediates repression of cartilage specific genes in human chondrocytes. AB - The role of histone deacetylase 1 and 2 (HDAC1 and HDAC2) in regulating cartilage specific gene expression was explored in primary human chondrocytes. HDAC1 and HDAC2 protein levels were elevated in chondrocytes from osteoarthritic patients, consistent with a down-regulation of some cartilage marker genes. When expressed in these cells, HDAC1 and HDAC2 repressed aggrecan and collagen 2(alpha1) expression but differed in their repression of collagen 9(alpha1), collagen 11(alpha1), dermatopontin, and cartilage oligomeric matrix protein (COMP). To identify the basis of these differences between HDAC1 and HDAC2, their carboxy terminal domains (CTDs) were deleted, which led to proteins that retained enzymatic activity but were unable to repress cartilage gene expression. Further, exchange of the CTDs between HDAC1 and HDAC2 led to proteins that were enzymatically active but displayed altered target gene specificity, indicating that these CTDs can function independently of HDAC enzymatic activity, to target the HDACs to specific genes. The Snail transcription factor was identified as a mediator of HDAC1 and HDAC2 repression of the collagen 2(alpha1) gene, via its interaction with the HDAC1 and 2 CTDs. The data indicate that the CTD serves a novel function within HDAC1 and HDAC2, to mediate repression of cartilage specific gene expression in human chondrocytes. PMID- 19561123 TI - Recent advances in PAMP-triggered immunity against bacteria: pattern recognition receptors watch over and raise the alarm. PMID- 19561125 TI - Dual mechanisms of sHA 14-1 in inducing cell death through endoplasmic reticulum and mitochondria. AB - HA 14-1 is a small-molecule Bcl-2 antagonist that promotes apoptosis in malignant cells, but its mechanism of action is not well defined. We recently reported that HA 14-1 has a half-life of only 15 min in vitro, which led us to develop a stable analog of HA 14-1 (sHA 14-1). The current study characterizes its mode of action. Because of the antiapoptotic function of Bcl-2 family proteins on the endoplasmic reticulum (ER) and mitochondria, the effect of sHA 14-1 on both organelles was evaluated. sHA 14-1 induced ER calcium release in human leukemic cells within 1 min, followed by induction of the ER stress-inducible transcription factor ATF4. Similar kinetics and stronger intensity of ER calcium release were induced by the sarcoendoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin, accompanied by similar kinetics and intensity of ATF4 induction. sHA 14-1 directly inhibited SERCA enzymatic activity but had no effect on the inositol triphosphate receptor. Evaluation of the mitochondrial pathway showed that sHA 14 1 triggered a loss of mitochondrial transmembrane potential (Delta psi m) and weak caspase-9 activation, whereas thapsigargin had no effect. (R)-4-(3 Dimethylamino-1-phenylsulfanylmethyl-propylamino)-N-{4-[4-(4'-chloro-biphenyl-2 ylmethyl)-piperazin-1-yl]-benzoyl}-3-nitrobenzenesulfonamide (ABT-737), a well established small-molecule Bcl-2 antagonist, rapidly induced loss of Delta psi m and caspase-9 activation but had no effect on the ER. The pan-caspase inhibitor N benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone had some protective effect on sHA 14-1-induced cell death. These collective results suggest a unique dual targeting mechanism of sHA 14-1 on the apoptotic resistance machinery of tumor cells that includes antiapoptotic Bcl-2 family proteins and SERCA proteins. PMID- 19561126 TI - A nondesensitizing kainate receptor point mutant. AB - Ionotropic glutamate receptor (iGluR) desensitization can be modulated by mutations that change the stability of a dimer formed by the agonist binding domain. Desensitization of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors can be blocked by a single point mutation (e.g., GluR2 L483Y) that stabilizes this dimer in an active conformation. In contrast, desensitization of kainate receptors can be slowed, but not blocked, by similar dimer interface mutations. Only covalent cross-linking via introduced disulfides has been previously shown to block kainate receptor desensitization completely. We have now identified an apparently nondesensitizing GluR6 point mutant (D776K) located at the apex of the ligand binding (S1S2) domain dimer interface. Asp776 is one of a cluster of four charged residues in this region that together mediate direct dimer interactions and contribute to the binding sites for one chloride and two sodium ions. Despite the localized +4 change in the net charge of the S1S2 dimer, the D776K mutation actually increased the thermodynamic stability of the dimer. Unlike GluR6 wild type, the D776K mutant is insensitive to external cations but retains sensitivity to external anions. We therefore hypothesize that the unexpected phenotype of this charge reversal mutation results from the substitution of the sodium ions bound within the dimer interface by the introduced lysine NH(3)(+) groups. The nondesensitizing D776K mutant provides insights into kainate receptor gating and represents a potentially useful new tool for dissecting kainate receptor function. PMID- 19561127 TI - The rolex and the hourglass: a simplified circadian clock in prochlorococcus? PMID- 19561128 TI - Production of 3-nitrosoindole derivatives by Escherichia coli during anaerobic growth. AB - When Escherichia coli K-12 is grown anaerobically in medium containing tryptophan and sodium nitrate, it produces red compounds. The reaction requires functional genes for trytophanase (tnaA), a tryptophan permease (tnaB), and a nitrate reductase (narG), as well as a natural drop in the pH of the culture. Mass spectrometry revealed that the purified chromophores had mass/charge ratios that closely match those for indole red, indoxyl red, and an indole trimer. These compounds are known products of chemical reactions between indole and nitrous acid. They are derived from an initial reaction of 3-nitrosoindole with indole. Apparently, nitrite that is produced from the metabolic reduction of nitrate is converted in the acid medium to nitrous acid, which leads to the nitrosation of the indole that is generated by tryptophanase. An nfi (endonuclease V) mutant and a recA mutant were selectively killed during the period of chromophore production, and a uvrA strain displayed reduced growth. These effects depended on the addition of nitrate to the medium and on tryptophanase activity in the cells. Unexpectedly, the killing of a tnaA(+) nfi mutant was not accompanied by marked increases in mutation frequencies for several traits tested. The vulnerability of three DNA repair mutants indicates that a nitrosoindole or a derivative of a nitrosoindole produces lethal DNA damage. PMID- 19561130 TI - The Acinetobacter baylyi Hfq gene encodes a large protein with an unusual C terminus. AB - In gammaproteobacteria the Hfq protein shows a great variation in size, especially in its C-terminal part. Extremely large Hfq proteins consisting of almost 200 amino acid residues and more are found within the gammaproteobacterial family Moraxellaceae. The difference in size compared to other Hfq proteins is due to a glycine-rich domain near the C-terminal end of the protein. Acinetobacter baylyi, a nonpathogenic soil bacterium and member of the Moraxellaceae encodes a large 174-amino-acid Hfq homologue containing the unique and repetitive amino acid pattern GGGFGGQ within the glycine-rich domain. Despite the presence of the C-terminal extension, A. baylyi Hfq complemented an Escherichia coli hfq mutant in vivo. By using polyclonal anti-Hfq antibodies, we detected the large A. baylyi Hfq that corresponds to its annotated size indicating the expression and stability of the full protein. Deletion of the complete A. baylyi hfq open reading frame resulted in severe reduction of growth. In addition, a deletion or overexpression of Hfq was accompanied by the loss of cell chain assembly. The glycine-rich domain was not responsible for growth and cell phenotypes. hfq gene localization in A. baylyi is strictly conserved within the mutL-miaA-hfq operon, and we show that hfq expression starts within the preceding miaA gene or further upstream. PMID- 19561129 TI - Metabolic flux analysis of Escherichia coli creB and arcA mutants reveals shared control of carbon catabolism under microaerobic growth conditions. AB - Escherichia coli has several elaborate sensing mechanisms for response to availability of oxygen and other electron acceptors, as well as the carbon source in the surrounding environment. Among them, the CreBC and ArcAB two-component signal transduction systems are responsible for regulation of carbon source utilization and redox control in response to oxygen availability, respectively. We assessed the role of CreBC and ArcAB in regulating the central carbon metabolism of E. coli under microaerobic conditions by means of (13)C-labeling experiments in chemostat cultures of a wild-type strain, DeltacreB and DeltaarcA single mutants, and a DeltacreB DeltaarcA double mutant. Continuous cultures were conducted at D = 0.1 h(-1) under carbon-limited conditions with restricted oxygen supply. Although all experimental strains metabolized glucose mainly through the Embden-Meyerhof-Parnas pathway, mutant strains had significantly lower fluxes in both the oxidative and the nonoxidative pentose phosphate pathways. Significant differences were also found at the pyruvate branching point. Both pyruvate formate lyase and the pyruvate dehydrogenase complex contributed to acetyl coenzyme A synthesis from pyruvate, and their activity seemed to be modulated by both ArcAB and CreBC. Strains carrying the creB deletion showed a higher biomass yield on glucose compared to the wild-type strain and its DeltaarcA derivative, which also correlated with higher fluxes from building blocks to biomass. Glyoxylate shunt and lactate dehydrogenase were active mainly in the DeltaarcA strain. Finally, it was observed that the tricarboxylic acid cycle reactions operated in a rather cyclic fashion under our experimental conditions, with reduced activity in the mutant strains. PMID- 19561131 TI - In vivo domain-based functional analysis of the major sporulation sensor kinase, KinA, in Bacillus subtilis. AB - Sensor histidine kinases are widely used by bacteria to detect and respond to environmental signals. In Bacillus subtilis, KinA is a major kinase providing phosphate input to the phosphorelay that activates the sporulation pathway upon starvation via the phosphorylated Spo0A transcription factor. KinA contains three PAS domains in its amino-terminal sensor domain, which appear to be involved in the sensing of an unidentified sporulation signal(s) produced upon starvation. Prior biochemical studies have suggested that KinA forms a homodimer as a functional enzyme and that the most amino-terminal PAS domain (PAS-A) plays an important role in sensing the signal(s) to activate an ATP-dependent autophosphorylation reaction to a histidine residue. To analyze the structure and function of the kinase in vivo, we have used a strain in which the synthesis of KinA is under the control of an isopropyl-beta-d-thiogalactopyranoside (IPTG) inducible promoter. In vivo functional studies in combination with domain-based deletion analysis show that the cytosolic KinA forms a homo-oligomer as an active form under both nutrient-rich and nutrient-depleted conditions via its amino- and carboxyl-terminal domains independently. Furthermore, we found that a mutant in which the PAS-A domain was deleted was still able to induce sporulation at a wild type level irrespective of nutrient availability, suggesting that PAS-BC domains are sufficient to maintain the kinase activity. Based on these results, we propose that the primary role of the amino-terminal sensor domain is to form a stable complex as a functional kinase, but possibly not for the binding of an unidentified sporulation signal(s). PMID- 19561133 TI - Electron microscopy of octacalcium phosphate in the dental calculus. AB - The purpose of this study was to morphologically demonstrate the presence of octacalcium phosphate in the dental calculus by judging from the crystal lattice image and its rapid transformation into apatite crystal, as part of our serial studies on biomineral products. We also aimed to confirm whether the physical properties of octacalcium phosphate are identical with those of the central dark lines observed in crystals of ordinary calcifying hard tissues. Electron micrographs showed that crystals of various sizes form in the dental calculus. The formation of each crystal seemed to be closely associated with the organic substance, possibly originating from degenerated microorganisms at the calcification front. Many crystals had an 8.2-A lattice interval, similar to that of an apatite crystal. Furthermore, some crystals clearly revealed an 18.7-A lattice interval and were vulnerable to electron bombardment. After electron beam exposure, this lattice interval was quickly altered to about half (i.e. 8.2 A), indicating structural conversion. Consequently, a number of apatite crystals in the dental calculus are possibly created by a conversion mechanism involving an octacalcium phosphate intermediate. However, we also concluded that the calcification process in the dental calculus is not similar to that of ordinary calcifying hard tissues. PMID- 19561134 TI - Criminal profiling as a plotting activity based on abductive processes. AB - In this article the authors analyze the nature and aims of criminal profiling from a theoretical point of view. The need to become increasingly "scientific" has given rise to the modern approaches of profiling, which have been particularly successful in cases of serial homicides and sex crimes, given that compulsive (perverse) acts, because of their ritual nature, have been described as being more easily foreseeable and presumably linkable to the psychological and even personal characteristics of a given criminal. On this basis, the authors analyze profiling from an epistemological point of view and show how, in the concrete activity of profiling, profilers depart from the "certainty" of the scientific models (those that are based on deductive-inductive processes); the epistemological basis of reasoning changes as there is no longer an induction deduction model but rather an abductive model (as conceived and explained by Peirce) in which the importance of plotting (the weaving of a narrative) becomes greater. PMID- 19561132 TI - The day-to-day process of stopping or reducing smoking: a prospective study of self-changers. AB - INTRODUCTION: Almost all descriptions of attempts to quit smoking have focused on what happens after an abrupt quit attempt and end once a smoker relapses. The current study examined the day-to-day process preceding a quit or reduction attempt in addition to the daily process after a failure to quit or reduce. METHODS: We recruited 220 adult daily cigarette smokers who planned to quit abruptly, to quit gradually, to reduce only, or to not change on their own. Participants called a voice mail system each night for 28 days to report cigarette use for that day and their intentions for smoking for the next day. No treatment was provided. RESULTS: Three main findings emerged: (a) The large majority of participants did not show a simple pattern of change but rather showed a pattern of multiple transitions among smoking, abstinence, and reduction over a short period of time; (b) most of those who reported an initial goal to quit abruptly actually reduced; and (c) daily intentions to quit strongly predicted abstinence, while daily intentions to reduce weakly predicted reduction. DISCUSSION: We conclude that the day-to-day process of attempts to change smoking among nontreatment seekers is much more dynamic than previously thought. This suggests that extended treatment beyond initial lapses and relapses and during postcessation reduction may be helpful. PMID- 19561135 TI - Perceptions and coping with punishment: how registered sex offenders respond to stress, internet restrictions, and the collateral consequences of registration. AB - This study examines how registered sex offenders (RSOs) experience, respond to, and attribute stress regarding sex offender registration and notification process and policies. In addition, the frequency and reasons for Internet access is assessed, with a focus on how new legislation in New Jersey (P.L. 2007, C.219) limits or blocks such access. Drawing on survey data from a random sample of 1,000 RSOs in New Jersey, responses from 107 RSOs show significant levels of stress, significant losses due to restrictions on Internet access, and coping methods associated with higher and lower stress levels. The most significant loss reported by the sample related to employment search difficulties. Factors associated with increased levels of stress include using self-distraction for coping and not accepting the situation. Surprisingly, being forced to move because of financial reasons is associated with lower levels of stress. PMID- 19561136 TI - Addition of growth hormone to gonadotrophins in ovarian stimulation of poor responders treated by in-vitro fertilization: a systematic review and meta analysis. AB - BACKGROUND: Whether the addition of growth hormone (GH) can improve the probability of pregnancy in poor responders undergoing ovarian stimulation for in vitro fertilization (IVF) has been examined to date by several underpowered studies, which have not provided solid conclusions. METHODS: A computerized literature search in MEDLINE, EMBASE, CENTRAL and randomized controlled trial (RCT) registries was performed independently by two reviewers, aiming to identify RCTs that evaluated the following research question: does GH addition increase the probability of pregnancy in poor responders undergoing ovarian stimulation with gonadotrophin releasing hormone (GnRH) analogues and gonadotrophins for IVF? RESULTS: Six relevant RCTs were identified, including a total of 169 patients. GH addition significantly increased clinical pregnancy (rate difference: +16%, 95% CI: +4 to +28; fixed effects model) (number-needed-to-treat (NNT) = 6, 95% CI: 4 25) and live birth rates (rate difference: +17%, 95% CI: +5 to +30; fixed effects model) (NNT = 6; 95% CI: 3-20). Furthermore, GH addition was associated with a significantly higher proportion of patients reaching embryo transfer (rate difference: +22%, 95% CI: +7 to +36; fixed effects model). CONCLUSIONS: The present meta-analysis provides evidence that GH addition increases the probability of clinical pregnancy and live birth in poor responders undergoing ovarian stimulation with GnRH analogues and gonadotrophins for IVF. However, the total number of patients analyzed is small and thus further RCTs are warranted to prove or disprove this finding. PMID- 19561137 TI - Adiponectin deficiency: a model of pulmonary hypertension associated with pulmonary vascular disease. AB - Adiponectin (APN) is an adipocyte-derived factor that exists at high concentrations in serum and has anti-inflammatory and systemic vascular protective properties. In this study, we investigated the role of APN in pulmonary vascular homeostasis. We found that APN localizes to the luminal side of blood vessels in lung and acts in vitro to block TNF-alpha-induced E-selectin upregulation in pulmonary artery endothelial cells. Targeted deletion of the APN gene in mice leads to a vascular phenotype in lung characterized by E-selectin upregulation and age-dependent increases in perivascular inflammatory cell infiltration and pulmonary arterial pressures. Taken together, these findings demonstrate an important role for APN in lung vascular homeostasis and suggest that APN-deficient states may contribute to the pathogenesis of inflammatory pulmonary vascular disease and to the development of pulmonary hypertension. PMID- 19561138 TI - Dexamethasone and mifepristone increase retroviral infectivity through different mechanisms. AB - Using adapted retroviruses for gene delivery is a modern and powerful tool in biological research as well as a promising approach for gene therapy. An important limitation for the extensive use of retroviral vectors is the low infection rate in target cells such as pulmonary vascular endothelial cells due to the insufficient infectivity of standard retrovirus supernatants that can only be overcome by complicated methods of virus concentration. This paper describes two easy methods to augment target cell infectivity, first by increasing the retroviral titer in the medium collected from packaging cells by stimulation of viral propagation with dexamethasone, and second, by increasing target cell sensitivity to retroviral infection by the glucocorticoid receptor antagonist, mifepristone. Using this method, we increased the infectivity of pulmonary microvascular endothelial cells from 16% to 85%. We demonstrate that mifepristone increased the susceptibility of target cells to retroviruses without increasing the viral titer of the supernatant. Dexamethasone, but not mifepristone, increased expression of delivered proteins such as GFP that are important for early identification of infected cells. Each, or both step(s), may be included in a standard protocol for retrovirus propagation and infection of target cells. PMID- 19561139 TI - Animal models of asthma. AB - Studies in animal models form the basis for much of our current understanding of the pathophysiology of asthma, and are central to the preclinical development of drug therapies. No animal model completely recapitulates all features of the human disease, however. Research has focused primarily on ways to generate allergic inflammation by sensitizing and challenging animals with a variety of foreign proteins, leading to an increased understanding of the immunological factors that mediate the inflammatory response and its physiological expression in the form of airways hyperresponsiveness. Animal models of exaggerated airway narrowing are also lending support to the notion that asthma may represent an abnormality of the airway smooth muscle. The mouse is now the species of choice for asthma research involving animals. This presents practical challenges for physiological study because the mouse is so small, but modern imaging methodologies, coupled with the forced oscillation technique for measuring lung mechanics, have allowed the asthma phenotype in mice to be precisely characterized. PMID- 19561141 TI - Effects of pre- and periconceptional undernutrition on arterial function in adult female sheep are vascular bed dependent. AB - The nutritional environment during development and even prior to conception may contribute to cardiovascular risk. In mature adult female sheep, we investigated the effect of preconceptional and periconceptional maternal nutritional restriction on the vascular reactivity of arteries from four vascular beds supplying the heart, thorax, kidney and hindlimb. Welsh Mountain ewes received 100% of nutrient requirements throughout gestation (control group, C, n = 18), or 50% of nutrient requirements for 30 days prior to conception (preconceptional group, PRE, n = 20) or for 15 days either side of conception (periconceptional group, PERI, n = 31) and 100% thereafter. In 3.5-year-old female offspring, the left anterior descending coronary (LAD), left internal thoracic (LITA), right renal and second and third order femoral arteries were dissected and their reactivity was assessed by organ bath or wire myography. Vasoconstrictor responses were greater in both LAD and LITA from PERI offspring compared with C (P < 0.01), while vasoconstriction was unaffected by maternal diet in arteries from the renal and femoral circulations (P = n.s.). Endothelium-dependent and independent vasodilatation was attenuated in third order femoral arteries of PRE and PERI groups compared with C (P < 0.05). Endothelium-independent vasodilatation was attenuated in both the LAD and renal arteries in the PERI group compared with C (P < 0.05). These data show that moderate maternal undernutrition either prior to or around conception affects vascular function of adult offspring. The effect depends on the timing of the insult, but also on the vascular bed studied and vessel hierarchy in the vascular tree. PMID- 19561142 TI - Voluntary activation of human knee extensors measured using transcranial magnetic stimulation. AB - The aim of this study was to determine the applicability and reliability of a transcranial magnetic stimulation twitch interpolation technique for measuring voluntary activation of a lower limb muscle group. Cortical voluntary activation of the knee extensors was determined in nine healthy men on two separate visits by measuring superimposed twitch torques evoked by transcranial magnetic stimulation during isometric knee extensions of varying intensity. Superimposed twitch amplitude decreased linearly with increasing voluntary torque between 50 and 100% of mean maximal torque, allowing estimation of resting twitch amplitude and subsequent calculation of voluntary activation. There were no systematic differences for maximal voluntary activation within day (mean +/- s.d. 90.9 +/- 6.2 versus 90.7 +/- 5.9%; P = 0.98) or between days (90.8 +/- 6.0 versus 91.2 +/- 5.7%; P = 0.92). Systematic bias and random error components of the 95% limits of agreement were 0.23 and 9.3% within day versus 0.38 and 7.5% between days. Voluntary activation was also determined immediately after a 2 min maximal voluntary isometric contraction; in four of these subjects, voluntary activation was determined 30 min after the sustained contraction. Immediately after the sustained isometric contraction, maximal voluntary activation was reduced from 91.2 +/- 5.7 to 74.2 +/- 12.0% (P < 0.001), indicating supraspinal fatigue. After 30 min, voluntary activation had recovered to 85.4 +/- 8.8% (P = 0.39 versus baseline). These results demonstrate that transcranial magnetic stimulation enables reliable measurement of maximal voluntary activation and assessment of supraspinal fatigue of the knee extensors. PMID- 19561140 TI - The protease inhibitor UCF-101 ameliorates streptozotocin-induced mouse cardiomyocyte contractile dysfunction in vitro: role of AMP-activated protein kinase. AB - 5-[5-(2-Nitrophenyl) furfuryliodine]-1,3-diphenyl-2-thiobarbituric acid (UCF-101) is a protease inhibitor which was reported to protect against ischaemic heart damage and apoptosis. This study evaluated the impact of UCF-101 on steptozotocin (STZ)-induced diabetic cardiomyocyte dysfunction. Adult FVB mice were made diabetic with a single injection of STZ (200 mg kg(1)). Two weeks after STZ injection, cardiomyocytes from control and STZ-treated mice were isolated and treated with UCF-101 (20 mum for 1 h). Cardiomyocyte contractile properties were analysed, including peak shortening (PS), maximal velocity of shortening/relengthening (+/-dL/dt), time to PS (TPS) and time to 90% relengthening (TR(90)). Steptozotocin-induced diabetes depressed PS and +/-dL/dt and prolonged TPS and TR(90) in cardiomyocytes, all of which were significantly alleviated by UCF-101. Immunoblotting analysis showed that UCF-101 significantly alleviated STZ-induced loss of phospholamban phosphorylation without affecting sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a) and phospholamban. Steptozotocin reduced AMP-activated protein kinase (AMPK) phosphorylation at Thr172 of the catalytic subunit without affecting total AMPK expression, which was restored by UCF-101. Short-term exposure to UCF-101 did not change the expression of X-linked inhibitor of apoptosis protein (XIAP) and Omi stress regulated endoprotease, high temperature requirement protein A2 (Omi/HtrA2), favouring an apoptosis-independent mechanism. Both the AMPK activator resveratrol and the antioxidant N-acetylcysteine mimicked the UCF-101-induced beneficial effect in STZ-induced diabetic cardiomyocytes. In addition, UCF-101 promoted the phosphorylation of p38 mitogen-activated protein kinases and c-Jun N-terminal kinase (JNK) after 15 min of incubation, while it failed to affect the phosphorylation of extracellular signal-regulated kinase (ERK) and glycogen synthase kinase-3beta (GSK-3beta) within 120 min in H9C2 myoblasts. Taken together, these results indicate that UCF-101 protects against STZ-induced cardiomyocyte contractile dysfunction, possibly via an AMPK-associated mechanism. PMID- 19561143 TI - Associations of the limb fat to trunk fat ratio with markers of cardiometabolic risk in elderly men and women. AB - BACKGROUND: The ratio of limb fat to trunk fat (LF/TF) is associated with markers of cardiometabolic risk in elderly men and women. It is unknown if LF/TF is associated with cardiometabolic risk beyond that explained by LF and TF independently. METHODS: Participants included abdominally obese men (n = 58) and women (n = 78) between 60 and 80 years of age. Regional adiposity was quantified using magnetic resonance imaging. Insulin resistance, fasting glucose, high density lipoprotein (HDL) cholesterol, plasma triglycerides, and adiponectin were determined using standard procedures. RESULTS: After control for potential confounders, TF was positively associated with fasting glucose, insulin resistance, and plasma triglycerides and negatively associated with HDL cholesterol and adiponectin (p < or = .05). These associations were strengthened after further control for LF (p < .05), with the exception of adiponectin in men (p > .05). After control for potential confounders, LF was negatively associated with adiponectin in men (p < .05) but not with any other marker of cardiometabolic risk (p > .05). After further control for TF, LF was negatively associated with plasma triglycerides and positively associated with HDL cholesterol in both genders combined (p < .05) and with adiponectin in women (p < .05) but not in men (p > .05). LF/TF was not associated with any marker of cardiometabolic risk after control for LF and TF. CONCLUSION: These results suggest that it is the absolute, rather than relative, amounts of LF and TF that have the greatest influence on cardiometabolic risk in elderly men and women. PMID- 19561144 TI - Previous hepatitis a virus infection is related to slower psychomotor speed in elderly adults. AB - BACKGROUND: Patients with chronic viral hepatitis are at a higher risk for cognitive dysfunction. Little is known about the association between hepatitis A virus (HAV) infection and cognitive function. METHODS: From the National Health and Nutrition Examination Survey, 1999-2002, we selected study participants (> or =60 years, n = 1,529) without hepatitis B, C, or D virus infection; without previous hepatitis A vaccination; and without abnormal liver function. HAV seropositive participants represented people with previous HAV infection. Psychomotor speed and executive functioning domain of cognitive function were measured by the Digit Symbol Substitution Test (DSST). RESULTS: HAV-seropositive participants had lower DSST scores than HAV-seronegative participants (weighted mean, 44.4 vs 53.9, p < .001). We designated HAV-seronegative participants as the reference group. Univariate analysis demonstrated that the weighted beta coefficient of DSST score was -9.55 (95% confidence interval [CI] -9.57 to -9.54, p < .001) for the HAV-seropositive participants. In a multivariable model, the weighted adjusted beta coefficient of DSST score was -2.48 (95% CI -2.49 to 2.46, p < .001) for the HAV-seropositive participants. CONCLUSION: HAV seropositivity is associated with slower psychomotor speed among the U.S. community-dwelling elders. PMID- 19561146 TI - A cross-sectional and longitudinal study of the relationship between walking speed and cognitive function in community-dwelling elderly people. AB - BACKGROUND: Previous reports have shown links between cognitive function and physical performance in the elderly people, but it is unclear whether some specific cognitive domains are more strongly associated with measures of physical function such as walking speed. We investigated cross-sectional and longitudinal relationships between performance in five cognitive tests and walking speed among community-dwelling elderly people in the Dijon center (France) of the Three-City Study. METHODS: At baseline, 3,769 participants aged 65-85 years had measurements of 6-m walking speed, global cognition (Mini-Mental State Examination), verbal fluency (Isaacs Set Test [IST]), psychomotor speed (Trail Making Test part A [TMT A]), executive function (TMT part B), and memory (Benton Visual Retention Test). After a mean follow-up of 7 years, walking speed was again measured in 1,732 of these participants. RESULTS: In cross-sectional analyses, slower maximum walking speed (MWS) at baseline was significantly associated with poorer performance in each cognitive test. The association was stronger with TMT-A (beta [SE] = -.127 [0.014], p < .0001) and IST (beta [SE] = .120 [0.014], p < .0001) than with the other tests. Only TMT-A (beta [SE] = -.053 [0.021], p = .01) and IST (beta [SE] = .063 [0.022], p = .004) were associated with the degree of MWS decline over time. CONCLUSIONS: This study shows both cross-sectional and longitudinal associations between cognition and walking speed among community-dwelling elderly people. Poorer verbal fluency and slower psychomotor speed were more specifically associated with slower baseline MWS and with a stronger decline in MWS over time. PMID- 19561147 TI - A comparison of the activities of lacticin 3147 and nisin against drug-resistant Staphylococcus aureus and Enterococcus species. AB - OBJECTIVES: Our goal was to compare the activities of lacticin 3147 and nisin, two of the most well characterized lantibiotics, against antibiotic-resistant staphylococci and enterococci. METHODS: We determined the MICs of lacticin 3147 and nisin for 20 strains of methicillin-resistant Staphylococcus aureus (MRSA), 20 strains of vancomycin-resistant enterococci (VRE), 6 strains of S. aureus with intermediate resistance to vancomycin (VISA), 5 strains of heterogeneous vancomycin-intermediate S. aureus (hVISA) and 4 strains of S. aureus that are susceptible to methicillin. RESULTS: Lacticin 3147 displayed potent activity against VRE with MIC values between 1.9 and 7.7 mg/L, and varying levels of activity against S. aureus strains (MRSA, 1.9-15.4 mg/L; laboratory strains, >or=15.4 mg/L; hVISA, 15.4-30.9 mg/L; VISA, >or=61.8 mg/L). Nisin was more active against the S. aureus strains in general (MRSA and laboratory strains, 0.5-4.1 mg/L; VISA and hVISA, 2 to >or=8.3 mg/L), but was less effective than lacticin 3147 against VRE (2 to >or=8.3 mg/L). CONCLUSIONS: Nisin is more effective against S. aureus whereas lacticin 3147 possesses greater potency against VRE. The modifications responsible for the vancomycin-resistant phenotypes of hVISA and VISA strains also provide protection against the two lantibiotics. PMID- 19561145 TI - Hormone therapy and skeletal muscle strength: a meta-analysis. AB - BACKGROUND: Our objective was to perform a systematic review and meta-analysis of the research literature that compared muscle strength in postmenopausal women who were and were not on estrogen-based hormone therapy (HT). METHODS: Twenty-three relevant studies were found. Effect sizes (ESs) were calculated as the standardized mean difference, and meta-analyses were completed using a random effects model. RESULTS: HT was found to result in a small beneficial effect on muscle strength in postmenopausal women (overall ES = 0.23; p = .003) that equated to an approximately 5% greater strength for women on HT. Among the 23 studies, various muscle groups were assessed for strength, and those that benefitted the most were the thumb adductors (ES = 1.14; p < .001). Ten studies that compared muscle strength in rodents that were and were not estradiol deficient were also analyzed. The ES for absolute strength was moderate but not statistically significant (ES = 0.44; p = .12), whereas estradiol had a large effect on strength normalized to muscle size (ES = 0.66; p = .03). CONCLUSION: Overall, estrogen-based treatments were found to beneficially affect strength. PMID- 19561148 TI - Synergistic effects of aminoglycosides and fosfomycin on Pseudomonas aeruginosa in vitro and biofilm infections in a rat model. AB - OBJECTIVES: To study the in vitro and in vivo efficacy of aminoglycosides against Pseudomonas aeruginosa, either alone or in combination with fosfomycin. METHODS: Using an in vitro study to assess inhibition of the growth of P. aeruginosa, MIC(90) and MIC(50) values of amikacin, gentamicin, netilmicin, tobramycin and isepamicin were determined, either alone or in combination with fosfomycin, and then the fractional inhibitory concentration index was calculated. In the biofilm infected rat model, the efficacy and effects of treatment with isepamicin and fosfomycin on infection were studied. RESULTS: The combinations of amikacin and fosfomycin or isepamicin and fosfomycin showed the most significant synergistic effects against P. aeruginosa as compared with other treatments. In the biofilm infected rat model, as a single agent, neither isepamicin nor fosfomycin reduced C-reactive protein level and numbers of white blood cells, or reduced the colony counts of the bacteria from both tissue and silica gel tubes. However, the combination of these two agents resulted in a good therapeutic effect. CONCLUSIONS: Combination of aminoglycosides and fosfomycin not only showed a positive effect in vitro but also improved the therapeutic effect in a biofilm infected rat model. This offers an effective treatment strategy against some therapy-resistant infections. PMID- 19561149 TI - Genetic interactions between the donor and the recipient for susceptibility to acute rejection in kidney transplantation: polymorphisms of CCR5. AB - BACKGROUND: Acute rejection (AR) contributes to the development of chronic allograft nephropathy that is the major cause of graft failure. We analyzed the 59029G>A polymorphism and an internal 32 bp deletion (CCR5 32) of CCR chemokine receptor 5 (CCR5) Delta and tried to prove the hypothesis that genetic interactions between the donor and the recipient influence the development of AR. METHODS: We detected genetic polymorphisms by the TaqMan(R) method and by sizing PCR amplicons (n = 486). The primary outcomes were early acute rejection (EAR) and repeated early acute rejection (RR). We defined EAR as the occurrence of a biopsy-proven AR within 3 months after transplantation. RESULTS: The development of EAR was dependent on the number of A alleles in recipients and showed a dose response relationship (P = 0.002). When we combined the number of A alleles in both donor and recipient, episodes of EAR and RR were more prevalent as the allelic number increased (A allelic number 0 & 1, 2 versus 3 & 4, P = 0.048; 0 & 1 versus 3 & 4, P = 0.006). Statistical significance was preserved after multivariate analysis of sex, HLA mismatch and type of donor with the recipient's age as the continuous term. Also, graft survival was different according to the presence of the A allele, i.e. recipients carrying A allele (+) grafts showed poor graft survival (P = 0.008 by a log-rank test). Again, the number of A alleles affected graft survival as the recipients who carried more A alleles had poor graft survival (A allele number 0 & 1 versus 2 versus 3 & 4, P = 0.011; 0 & 1 versus 3 & 4, P = 0.08; 0 & 1 versus 2, P = 0.002; by a log-rank test). All of the participants were wild-type homozygotes for CCR5Delta32. CONCLUSIONS: The A allele of CCR5 59029G>A was a risk factor for EAR and RR. As the number of A alleles increased, episodes of EAR were more frequently observed. PMID- 19561150 TI - Post-dilution haemodiafiltration and low-flux haemodialysis have dissimilar effects on platelets: a side study of CONTRAST. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death in patients with end-stage renal disease (ESRD). Platelet (PLT) dysfunction, which is a well-known phenomenon in advanced chronic renal failure, corresponds positively with CVD in these patients. The accumulation of retained uraemic toxins might play an important role in this respect. During haemodialysis (HD), both an increase in the expression of the platelet (PLT) cell surface molecule P selectin (CD62p) and the release of intra-granular substances, such as platelet factor 4 (PF4) and ss-thromboglobulin (BTG), have been described. As the removal of uraemic toxins is superior during haemodiafiltration (HDF), this form of treatment may have quite another impact on PLTs than HD. METHODS: Nineteen chronic HD patients who were treated with low-flux HD for at least 2 months were included in the Dutch CONvective TRAnsport STudy (CONTRAST). After randomization, 10 patients continued low-flux HD and 9 patients switched to post-dilution HDF. The present study describes various parameters of PLT activation and degranulation at baseline (during HD) and after 3 months (during HDF) in the latter group of patients. At both time points, multiple blood samples were drawn. During the first 30 min of treatment, differences over the extracorporeal circuit (ECC) were calculated by taking samples from both afferent (arterial) and efferent (venous) lines. Correlations between various parameters were calculated in the total group of patients after 3 months. RESULTS: Immediately after the start of HD, PLT counts dropped over the ECC. During HDF, PLT counts decreased even more and reached a nadir at t30. CD62p expression increased early during HD and returned to baseline thereafter. During HDF, these changes were more pronounced and more protracted. With respect to degranulation, rather dissimilar results were obtained. During HD, both PF4 and BTG increased over time, whereas during HDF, PF4 increased but BTG did not change. Haemoconcentration and transmembrane pressure (TMP) within the dialyser were, respectively, approximately 10 and 3x higher during HDF than during HD. There was a striking correlation between the changes in haemoconcentration and the changes in both PLT counts and CD62p over the ECC. SUMMARY AND CONCLUSIONS: PLT activation, as measured by the expression of CD62p, was more pronounced and more protracted during HDF than during HD. During HDF, PLTs were trapped abundantly within the ECC, not only after first passage, but also thereafter. The degranulation product BTG increased during HD, but did not change during HDF. These observations may well be explained by the greater haemoconcentration and/or higher TMP during HDF on the one hand, and superior convective transport at the other. Whether the potential harmful effects of enhanced PLT activation are counterbalanced by the beneficial effects of an increased convective transport of degranulation products remains to be established. PMID- 19561151 TI - Associations of VEGF and its receptors sVEGFR-1 and -2 with cardiovascular disease and survival in prevalent haemodialysis patients. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) was recently shown to predict survival in prevalent haemodialysis patients. Soluble VEGF receptors (sVEGFR)-1 and -2 are circulating endogenous modulators of VEGF activity. We thus studied the relationship between sVEGFR-1 and -2 and survival in a cohort of prevalent haemodialysis (HD) patients. METHODS: Components of the VEGF system were measured (ELISAs) in 185 prevalent HD patients and levels related to clinical characteristics, biochemical markers and survival. The patients were followed up prospectively for a median 31 (20-37) months. RESULTS: While ischaemic heart disease was independently associated with a lower sVEGFR-2 (OR = 2.75, P = 0.02), sVEGFR-1 was positively associated with IL-6 (rho = 0.22, P = 0.003) and white blood cell count (rho = 0.22, P = 0.002). In survival analysis, the patients with a high sVEGFR-1 level had a higher all-cause mortality (Kaplan Meier Chi-Square = 5.6, P = 0.02) and a higher adjusted mortality risk (Cox HR = 1.93, P = 0.009) than those with low levels. CONCLUSION: In the first clinical study of sVEGFR-1 and -2 in CKD, we found novel associations between the sVEGFRs and cardiac disease. This may be of clinical importance, as a high sVEGFR-1 was an independent risk factor for all-cause mortality. PMID- 19561152 TI - Autoregulation of glomerular filtration rate during spironolactone treatment in hypertensive patients with type 1 diabetes: a randomized crossover trial. AB - BACKGROUND: Autoregulation of GFR, i.e. maintenance of relative constancy of GFR despite variations in mean arterial pressure (MAP) >80 mmHg, is impaired in diabetic kidney disease; furthermore, some antihypertensive drugs may jeopardize autoregulation. The aim of our study was to establish if spironolactone affects the ability to autoregulate GFR. METHODS: Sixteen hypertensive type 1 diabetic patients with persistent normoalbuminuria (presumed normal autoregulation) completed this randomized, double-masked, crossover trial. After a 4-week wash out period, patients received spironolactone 25 mg o.d. and matched placebo for 4 weeks in random order. After each treatment period, the ability to autoregulate GFR was determined by measuring GFR ((51)Cr-EDTA clearance) before (basal) and after acute blood pressure reduction by intravenous injection of clonidine. RESULTS: During placebo, the mean (SE) basal GFR was 115 (5) ml/min/1.73 m(2) and the BP was 146 (4)/81 (2) mmHg corresponding to a MAP of 103 (2) mmHg. Spironolactone did not significantly reduce GFR or BP. Injection of clonidine induced a significant reduction in the MAP of 17 (2) and 19 (1) mmHg during placebo and spironolactone treatment, respectively, and an overall reduction in GFR of 11 and 15 ml/min/1.73 m(2) (both comparisons NS between treatment periods). Signs of impaired autoregulation were present in nine patients during placebo and in nine patients during spironolactone treatment. Relative changes in GFR on placebo treatment correlated with diabetes duration (R = 0.67, P < 0.01) but were not related to duration of hypertension, baseline BP, GFR, HbA1c or to changes in BP. CONCLUSION: Spironolactone did not change the overall ability to autoregulate GFR in 16 hypertensive type 1 diabetic patients with normoalbuminuria. Our data are suggestive that the ability to autoregulate GFR is gradually impaired with increasing diabetes duration, a phenomenon not previously described in normoalbuminuric patients. PMID- 19561154 TI - Inter-comparison study of the population dose due to gamma-radiation in the coast of North Sinai between Rafah and Bir El-Abd areas. AB - This study compares the external dose due to the gamma-ray emitting radionuclides in various areas in North Sinai, Egypt. The whole area was divided into 10 regions. The average absorbed dose rates were evaluated for each region. It was found that Zaranik-protected area and Al-Massaid have the highest values of 72.7 and 57.2 nGy h(-1), respectively. The corresponding values of the remaining regions were <23 nGy h(-1). The mean annual effective dose equivalents for the four largest cities Rafah, El-Sheikh Zuwaied, Al-Arish and Bir El-Abd were 20.8, 18.8, 57.4 and 14.0 microSv, respectively. The results are compared with those from different areas in Egypt and in various countries. PMID- 19561153 TI - In vivo up-regulation of kinin B1 receptors after treatment with Porphyromonas gingivalis lipopolysaccharide in rat paw. AB - It has been demonstrated that kinin B(1) receptors are highly up-regulated under several stressful stimuli, such as infection. However, there is no evidence indicating whether Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) might lead to B(1) receptor up-regulation. In this study, we demonstrate that Pg-LPS injection into the rat paw resulted in a marked functional up-regulation of B(1) receptors (as measured by an increase of B(1) receptor-induced edema), which was preceded by a rapid rise in B(1) receptor mRNA expression. The local administration of Pg-LPS also resulted in a prominent production of the proinflammatory cytokine tumor necrosis factor alpha (TNF-alpha), followed by an increase of neutrophil influx; both events were observed at periods before B(1) receptor induction. The functional and molecular Pg-LPS-elicited B(1) receptor up regulation was significantly reduced by the glucocorticoid dexamethasone (0.5 mg/kg s.c.), and to a lesser extent by the chimeric anti-TNF-alpha antibody infliximab (1 mg/kg s.c.). Of high relevance, we show for the first time that a single administration of the proresolution lipid mediator (5S,12R,18R)-trihydroxy 6Z,8E,10E,14Z,16E-eicosapentaenoic acid (resolvin E1; 300 ng/rat i.p.) was able to markedly down-regulate Pg-LPS-driven B(1) receptor expression, probably by inhibiting TNF-alpha production and neutrophil migration. Collectively, the present findings clearly suggest that Pg-LPS is able to induce the up-regulation of B(1) receptors through mechanisms involving TNF-alpha release and neutrophil influx, which are largely sensitive to resolvin E1. It is tempting to suggest that kinin B(1) receptors might well represent a pivotal pathway for the inflammatory responses evoked by P. gingivalis and its virulence factors. PMID- 19561155 TI - Rod phantom dimensions--does size matter? AB - The international standard ISO 4037-3:1999 recommends a cylindrical polymethyl methacrylate rod phantom of 300 mm length and 19 mm diameter for the calibration of extremity dosimeters worn on the finger. No tolerances are stated. This paper investigates the influence of the actual rod phantom dimensions on the calibration results of finger dosimeters. For this purpose, thermoluminescence detectors were attached to rod phantoms of different size and irradiated at the PTB reference X-ray fields. Rod phantoms with a diameter of (19 +/- 1) mm and a length of at least 100 mm can be considered to be equivalent for the calibration of finger dosimeters, since the variation in the calibration results is below the significance level of 1 %. The decrease in the response towards the edge of the phantom is negligible as long as the distance from the nearest edge exceeds 10 mm. PMID- 19561156 TI - Ultrasonographic evaluation of joint involvement in early rheumatoid arthritis in clinical remission: power Doppler signal predicts short-term relapse. AB - OBJECTIVES: This study aimed to evaluate the usefulness of a systematic musculoskeletal ultrasonographic (US) assessment in the detection of residual disease activity in patients with early RA who achieved clinical remission. METHODS: We prospectively studied 106 early RA patients receiving conventional DMARDs according to a disease activity score (DAS)-steered therapeutic protocol over a 24-month period. Standard evaluation included clinical, laboratory, functional and systematic (44 joints) US assessment. US indexes of grey scale (GS) and power Doppler (PD) synovitis were correlated with clinical evaluation, laboratory indexes and clinical outcome. Clinical remission was defined when DAS was <1.6 at two consecutive visits 3 months apart. RESULTS: US examination was significantly more sensitive than clinical examination, both in active disease and in remission. In patients with an active disease, both clinical and US indexes correlated with CRP, whereas in remission only PD still remained significantly correlated. In clinical remission, 95% of the patients showed residual GS synovitis, and 41% of them showed a positive PD signal. Positive PD signal, even in a single joint, resulted the main predictor of relapse within 6 months, both in univariable and multivariable logistic regression analysis. CONCLUSIONS: In a cohort of early RA patients treated with conventional DMARDs, US-GS can detect residual disease activity more sensitively than clinical examination both in active disease and in remission. Moreover, PD-positive synovial hypertrophy identifies an ongoing inflammation even during remission and predicts short-term relapse. PMID- 19561157 TI - Combination of TNF-RII, CYP1A1 and GSTM1 polymorphisms and the risk of Japanese SLE: findings from the KYSS study. AB - OBJECTIVES: Association of the polymorphisms of the genes, TNF receptor type II gene (TNF-RII), cytochrome P4501A1 gene (CYP1A1) and glutathione S-transferase M1 gene (GSTM1), with SLE was investigated. TNF-RII mediates inflammatory and immune response, whereas CYP1A1 and GSTM1 are involved in the metabolism of xenobiotics. These three genes are involved in the generation of reactive oxygen species (ROS), which play a critical role for autoimmune diseases. METHODS: A total of 152 SLE patients and 427 healthy individuals in a female Japanese population were enrolled in the study. Case-control studies were performed for the polymorphisms of these three genes. RESULTS: The carriers of TNF-RII 196R were at a significantly increased risk for SLE with odds ratio (OR) of 1.59 (95% CI = 1.01, 2.52). CYP1A1 3801C homozygotes had a significantly increased risk of SLE (OR = 2.47, 95% CI = 1.28, 4.78). On the other hand, GSTM1 null genotype was not associated with SLE risk. As for combination action of two loci, CYP1A1 3801C/GSTM1 null combination was more strongly associated with an increased risk of SLE (OR = 4.35; 95% CI = 1.76, 10.73). Moreover, TNF-RII 196M/CYP1A1 3801C/GSTM1 null genotype combination was most significantly associated with SLE (OR = 5.83; 95% CI = 2, 17.04). CONCLUSIONS: The individuals carrying two or more 'at-risk' genotypes of TNF-RII, CYP1A1 and GSTM1 had a significantly more increased risk for SLE compared with those having each 'at-risk' genotype. Combination of the risk genotypes will be important to more clearly identify the population at risk for SLE. PMID- 19561158 TI - Treatment of ankylosing spondylitis and extra-articular manifestations in everyday rheumatology practice. AB - The SpAs are a group of overlapping, chronic, inflammatory rheumatic diseases including AS, a chronic inflammatory disease primarily affecting the SI joints. In addition to inflammatory back pain, AS patients are also more likely to experience extra-articular manifestations belonging to the SpA concept which can affect the eyes, the gastrointestinal tract and the skin and other related inflammatory conditions. This review focuses on current progress in treatment options in SpA with special emphasis on extra-articular features. TNF inhibition has demonstrated effectiveness in the treatment of AS symptoms and all currently available anti-TNF agents appear to have similar efficacy. However, the efficacy of anti-TNF agents varies in the treatment of extra-articular manifestations and comorbidities. Analyses of trials of anti-TNF agents in patients with AS have revealed significant reductions in the incidence of flares of uveitis and IBD with infliximab and adalimumab (uveitis only) treatment but not with etanercept. All three anti-TNF agents (infliximab, adalimumab, etanercept) have demonstrated efficacy in psoriasis (not associated with AS). When evaluating as to which agent to use in the treatment of AS, an important consideration is the overall well being of the patient. This should include any additional inflammatory burden that manifests in other parts of the body, which may currently be subclinical. Based on current evidence, among TNF inhibitors, the monoclonal antibodies (infliximab and adalimumab) are more appropriate than etanercept if extra-articular manifestations or comorbid conditions are present or suspected. To date, infliximab appears to be the best studied agent with a wide spectrum of proven efficacy. PMID- 19561159 TI - Multipotent mesenchymal stromal cells and rheumatoid arthritis: risk or benefit? AB - Multipotent mesenchymal stromal cells (MSCs) have raised interest mainly because of cartilage/bone differentiation potential which is now partly eclipsed by their capacity to counteract inflammation and suppress host immune responses as well as to prevent fibrosis. MSCs have been identified within joint tissues including synovium, cartilage, subchondral bone, periosteum or adipose tissue. They are characterized by their phenotype and their ability to differentiate into three lineages, chondrocytes, osteoblasts and adipocytes. MSCs have also paracrine effects through the secretion of a number of cytokines and growth factors. This may explain the trophic effects that may be of therapeutic value for rheumatic diseases including OA and RA. On the other hand, MSCs have been associated with tumour growth. MSCs migrate to the tumour stroma, express chemokines involved in the attraction of carcinoma cells in metastasis. Indeed, the aim of this review is not only to focus on new potential therapeutic applications in osteo-articular diseases, but also to assess the potential risk of MSC-based cell therapy. PMID- 19561160 TI - Prevalence and prognostic implications of dysphagia in elderly patients with pneumonia. AB - BACKGROUND: oropharyngeal dysphagia is a common condition among the elderly but not systematically explored. OBJECTIVE: to assess the prevalence and the prognostic significance of oropharyngeal dysphagia among elderly patients with pneumonia. DESIGN: a prospective cohort study. SETTING: an acute geriatric unit in a general hospital. SUBJECTS: a total of 134 elderly patients (>70 years) consecutively admitted with pneumonia. METHODS: clinical bedside assessment of oropharyngeal dysphagia and aspiration with the water swallow test were performed. Demographic and clinical data, Barthel Index, Mini Nutritional Assessment, Charlson Comorbidity Index, Fine's Pneumonia Severity Index and mortality at 30 days and 1 year after admission were registered. RESULTS: of the 134 patients, 53% were over 84 years and 55% presented clinical signs of oropharyngeal dysphagia; the mean Barthel score was 61 points indicating a frail population. Patients with dysphagia were older, showed lower functional status, higher prevalence of malnutrition and comorbidities and higher Fine's pneumonia severity scores. They had a higher mortality at 30 days (22.9% vs. 8.3%, P = 0.033) and at 1 year of follow-up (55.4% vs. 26.7%, P = 0.001). CONCLUSIONS: oropharyngeal dysphagia is a highly prevalent clinical finding in elderly patients with pneumonia and is an indicator of disease severity in older patients with pneumonia. PMID- 19561161 TI - Abnormal Savda Munziq, an Herbal Preparation of Traditional Uighur Medicine, May Prevent 1,2-dimethylhydrazine-Induced Rat Colon Carcinogenesis. AB - The study tried to assess the chemoprotective effect of abnormal Savda Munziq (ASMq) on 1,2-dimethylhydrazine (DMH)-induced rat colon carcinogenesis. Male F344 rats were randomized into eight groups: Group 1 was served as control, no DMH injection was given and treated daily with normal saline. Rats in Groups 2-8 were given a single intraperitoneal injection of DMH (20 mg/kg body weight) at the beginning of the study. Group 2 was served as negative control, administered with normal saline until the end of the experiment after the single DMH injection. Groups 3-5 were served as pretreatment group, administered with ASMq ethanol extract at 400, 800 and 1600 mg/kg body weight, respectively, until the 45th day, continued by normal saline administration for another 45 days. Groups 6-8 were served as the treatment group, administered with normal saline for the first 45 days from the day of DMH injection, ASMq ethanol extract at three different doses to be administered until the end of the second 45th day. All rats were sacrificed at 91st day and the colons were analyzed for aberrant crypt foci (ACF) formation and crypt multiplicity. Results showed that ASMq ethanol extract reduced the number of ACF, AC and crypt multiplicity significantly (P < .05). It suggested that ASMq ethanol extract had chemoprotective effects on DMH-induced colon carcinogenesis, by suppressing the development of preneoplastic lesions, and probably exerted protection against the initiation and promotion steps of colon carcinogenesis. PMID- 19561162 TI - Hostility and minimal model of glucose kinetics in African American women. AB - OBJECTIVE: To explore the underlying physiology of hostility (HOST) and to test the hypothesis that HOST has a greater impact on fasting glucose in African American (AA) women than it does on AA men or white men or women, using an intravenous glucose tolerance test (IVGTT) and the minimal model of glucose kinetics. METHODS: A total of 115 healthy subjects selected for high or low scores on the 27 item Cook Medley HOST Scale underwent an IVGTT. Fasting nonesterified fatty acids (NEFA) levels were measured before the IVGTT. Catecholamine levels were measured 10 minutes into the IVGTT. RESULTS: Moderation by group (AA women versus others) of HOST was found for glucose effectiveness (Sg, p = .02), acute insulin response (AIRg, p = .02), and disposition index (DI, p = .02). AA women showed a negative association between HOST and both Sg (beta = -0.45, p = .04) and DI (beta = -0.49, p = .02), controlling for age and body mass index. HOST was also associated with changes in epinephrine (beta = 0.39, p = .05) and fasting NEFA (beta = 0.44, p = .02) in the AA women. Controlling for fasting NEFA reduced the effect of HOST on both Sg and DI. CONCLUSIONS: This study shows that HOST is related to decreased DI, a measure of pancreatic compensation for increased insulin resistance as well as decreased Sg, a measure of noninsulin-mediated glucose transport compared in AA women. These effects are partly mediated by the relationship of HOST to fasting NEFA. PMID- 19561164 TI - Chronic obstructive pulmonary disease patients with psychiatric disorders are at greater risk of exacerbations. AB - OBJECTIVE: To assess prospectively the impact of psychiatric disorders on risk for exacerbations. The course of chronic obstructive pulmonary disease (COPD) is punctuated by acute exacerbations. Although anxiety and mood disorders are common in patients with COPD, no studies have assessed prospectively the association between these disorders and exacerbations. METHODS: Psychiatric disorders were evaluated by a structured psychiatric interview in 110 patients (51% women, age (mean +/- standard deviation) = 66 +/- 8 years) with stable COPD and previous admission for exacerbations recruited from two outpatient clinics. Patients were followed for a mean of 2 years and both inpatient-treated (i.e., treated in the emergency department or hospital) and outpatient-treated (i.e., treated with medication in the patient's own environment) exacerbations were recorded. RESULTS: Independent of covariates, patients with psychiatric disorders exhibited a significantly higher weighted annual rate of exacerbations treated in an outpatient setting after adjustment for covariates (3 versus 2, p = .003) than patients without psychiatric disorders, but no difference in exacerbations treated in the inpatient setting. They were also at a higher risk for any (relative risk (RR) = 1.56, 95% Confidence Interval (CI) = 1.02-2.37) and outpatient (RR = 1.68, 95% CI = 1.08-2.59) exacerbations, but not inpatient exacerbations (RR = 1.36, 95% CI = 0.82-2.25). CONCLUSIONS: Patients with psychiatric disorders are at greater risk of exacerbations treated in an outpatient setting but not those treated in an inpatient setting. These outpatient-managed exacerbations account for a significant proportion of the healthcare burden for COPD, so interventions should target patients with psychiatric disorders to improve management of COPD. PMID- 19561165 TI - Brain activation associated with changes in heart rate, heart rate variability, and plasma catecholamines during rectal distention. AB - OBJECTIVE: To test the hypothesis that gut stimulation provokes autonomic arousal via activation of regional cerebral cortices. How the human brain processes interoceptive signals and forms initial autonomic arousal is one of the key questions to be answered in research on emotion. METHODS: Twelve healthy males participated in this study. A barostat bag was inserted in the rectum and intermittently inflated with 0, 20, or 40 mm Hg at random for 80 seconds. H(2)(15)O positron emission tomography (PET) of the brain, electrocardiography, and blood sampling for catecholamines were performed. Changes in regional cerebral blood flow were interpreted using statistical parametric mapping. RESULTS: Rectal distention with 40 mm Hg induced a significant increase in heart rate, low frequency (LF)/high frequency (HF) ratio of heart rate variability, and plasma adrenaline. Activated brain areas that were associated with increased heart rate during rectal distention were the right insula, right operculum, right dorsolateral prefrontal cortex, putamen, thalamus, periaqueductal gray, and cerebellum (p < .001, uncorrected), whereas those that were associated with an increased LF/HF ratio were the bilateral insula, putamen, thalamus, midbrain, pons, and cerebellum (p < .001, uncorrected). Activated brain areas that were associated with increased plasma adrenaline were the right insula, right orbitofrontal cortex, right parahippocampal gyrus, putamen, thalamus, periaqueductal gray, pons, and cerebellum (p < .001, uncorrected). CONCLUSION: Our results suggest that the right insula and the related body mapping regions may form the functional module of sympathetic arousal in response to gut stimulation. PMID- 19561163 TI - Manic/hypomanic symptom burden and cardiovascular mortality in bipolar disorder. AB - OBJECTIVES: To compare the risk for cardiovascular mortality between bipolar I and bipolar II subtypes and determine correlates of cardiovascular mortality. Bipolar disorder conveys an increased risk of cardiovascular mortality. METHODS: Participants with major affective disorders were recruited for the National Institute of Mental Health Collaborative Depression Study and followed prospectively for up to 25 years. A total of 435 participants met the diagnostic criteria for bipolar I (n = 288) or bipolar II (n = 147) disorder based on Research Diagnostic Criteria at intake and measures of psychiatric symptoms during follow-up. Diagnostic subtypes were contrasted by cardiovascular mortality risk using Cox proportional hazards regression. Affective symptom burden (the proportion of time with clinically significant manic/hypomanic or depressive symptoms) and treatment exposure were additionally included in the models. RESULTS: Thirty-three participants died from cardiovascular causes. Participants with bipolar I disorder had more than double the cardiovascular mortality risk of those with bipolar II disorder, after controlling for age and gender (hazard ratio = 2.35, 95% Confidence Interval = 1.04-5.33; p = .04). The observed difference in cardiovascular mortality between these subtypes was at least partially confounded by the burden of clinically significant manic/hypomanic symptoms which predicted cardiovascular mortality independent of diagnosis, treatment exposure, age, gender, and cardiovascular risk factors at intake. Selective serotonin uptake inhibitors seemed protective although they were introduced late in follow-up. Depressive symptom burden was not related to cardiovascular mortality. CONCLUSIONS: Participants with bipolar I disorder may face a greater risk of cardiovascular mortality than those with bipolar II disorder. This difference in cardiovascular mortality risk may reflect manic/hypomanic symptom burden. PMID- 19561166 TI - Licensure tests for special education teachers: how well they assess knowledge of reading instruction and mathematics. AB - To determine the extent to which knowledge of evidence-based reading instruction and mathematics is assessed on licensure tests for prospective special education teachers, this study drew on information provided by Educational Testing Service (ETS), the American Board for Certification of Teacher Excellence, and National Evaluation Systems (now Evaluation Systems group of Pearson). It estimated the percentage of test items on phonemic awareness, phonics, and vocabulary knowledge and on mathematics content. It also analyzed descriptions of ETS's tests of "principles of teaching and learning." Findings imply that prospective special education teachers should be required to take both a dedicated test of evidence based reading instructional knowledge, as in California, Massachusetts, and Virginia, and a test of mathematical knowledge, as in Massachusetts. States must design their own tests of teaching principles to assess knowledge of evidence based educational theories. PMID- 19561167 TI - Validating the location of fluorescent protein fusions in the endomembrane system. AB - Assessment of gene function generally requires knowledge of the sites of action of gene products. Several experimental approaches can provide relevant information, but all have their limitations and the potential for experimental artifact. In this article we focus on the endomembrane organelles and on the methods that can be used to validate the location of fluorescent protein fusions. We discuss the utility of redundant localization techniques, complementation of mutant phenotypes, and integration of localization data with expected biological function as methods to achieve consensus. We argue that no single piece of evidence is sufficient to address the issue, and that all approaches can reveal useful information about the true steady state location of a protein or about other aspects of its transport and dynamics. As ever, the critical point is the subjective interpretation one puts on each observation in light of the experimental conditions and other pertinent data. We illustrate these points with some successes and failures in our own work. PMID- 19561169 TI - Fancm-deficient mice reveal unique features of Fanconi anemia complementation group M. AB - The Fanconi anemia (FA) core complex member FANCM remodels synthetic replication forks and recombination intermediates. Thus far, only one FA patient with FANCM mutations has been described, but the relevance of these mutations for the FA phenotype is uncertain. To provide further experimental access to the FA-M complementation group we have generated Fancm-deficient mice by deleting exon 2. FANCM deficiency caused hypogonadism in mice and hypersensitivity to cross linking agents in mouse embryonic fibroblasts (MEFs), thus phenocopying other FA mouse models. However, Fancm(Delta2/Delta2) mice also showed unique features atypical for FA mice, including underrepresentation of female Fancm(Delta2/Delta2) mice and decreased overall and tumor-free survival. This increased cancer incidence may be correlated to the role of FANCM in the suppression of spontaneous sister chromatid exchanges as observed in MEFs. In addition, FANCM appeared to have a stimulatory rather than essential role in FANCD2 monoubiquitination. The FA-M mouse model presented here suggests that FANCM functions both inside and outside the FA core complex to maintain genome stability and to prevent tumorigenesis. PMID- 19561168 TI - Exploring the function-location nexus: using multiple lines of evidence in defining the subcellular location of plant proteins. AB - Defining the function of all proteins in an organism is one of the major objectives for biology in the coming decades. Here, we assess approaches used to determine subcellular protein location and discuss the relationship between protein location and function. It is important to recognize that targeting, accumulation, and the site of function are not necessarily interchangeable terms with respect to defining the location of a protein. Some proteins have tightly defined locations, whereas others have low specificity targeting and complex accumulation patterns. Location may be essential for function in some cases, but it may be much less important for other proteins. There is no single approach that can be considered entirely adequate for defining the in vivo location of all proteins. By combining approaches that assess targeting and accumulation of proteins, more confidence can be gained about localization. The strengths and weaknesses of different localization technologies are summarized, and some guidelines for performing combined targeting and accumulation assays are outlined. PMID- 19561170 TI - Protein kinase C gamma, a protein causative for dominant ataxia, negatively regulates nuclear import of recessive-ataxia-related aprataxin. AB - Spinocerebellar ataxia type 14 (SCA14) is an autosomal dominant disease caused by mutations in the gene encoding protein kinase C gamma (PKC gamma). We report an SCA14 family with a novel deletion of a termination-codon-containing region, resulting in a missense change and a C-terminal 13-amino-acid extension with increased kinase activity. Notably, one patient with a severe phenotype is the first homozygote for the mutation causing SCA14. We show the novel molecular consequences of increased kinase activities of mutants: aprataxin (APTX), a DNA repair protein causative for autosomal recessive ataxia, was found to be a preferential substrate of mutant PKC gamma, and phosphorylation inhibited its nuclear entry. The phosphorylated residue was Thr111, located adjacent to the nuclear localization signal, and disturbed interactions with importin alpha, a nuclear import adaptor. Decreased nuclear APTX increased oxidative stress-induced DNA damage and cell death. Phosphorylation-resistant APTX, kinase inhibitors, and antioxidants may be therapeutic options for SCA14. PMID- 19561171 TI - Genomic regulatory blocks in vertebrates and implications in human disease. AB - Despite a recent explosion in the production of vertebrate genome sequence data and large-scale efforts to completely annotate the human genome, we still have scant knowledge of the principles that built vertebrate genomes in evolution, and of genome architecture and its functional significance. We review approaches using bioinformatics, zebrafish transgenesis, and recent findings in the molecular basis of gene regulation and tie these in with mechanisms for the maintenance of long-range conserved synteny across all vertebrate genomes. Specifically, we discuss the recently discovered genomic regulatory blocks which we argue are principal units of vertebrate genome evolution and serve as the foundations onto which evolutionary innovations are built through sequence evolution and insertion of new cis-regulatory elements. We subsequently discuss how these arrangements relate to common human heritable diseases and their significance in disease causality. PMID- 19561172 TI - The contribution of qualitative research to the development of tailor-made community-based interventions in primary care: a review. AB - BACKGROUND: In recent years, a trend in the use of tailor-made approaches and pragmatic trial methodology for evaluating effectiveness has been visible in programs ranging from large-scale national health prevention campaigns to community-based initiatives. Qualitative research is used more often for tailoring interventions towards communities and/or local care practices. This article systematically reviews the contribution of qualitative research in developing tailor-made community-based interventions in primary care evaluated by means of the pragmatic trial methodology. METHODS: A systematic search of Pubmed/Medline and Embase revealed 33 articles. Using a literature mapping process, the articles were arranged according to the development phases identified in the MRC framework for the development of complex interventions to improve health. RESULTS: The review showed qualitative research is mainly used to provide insight into the contextual circumstances of the interventions' implementation, delivery and evaluation. To a lesser extent, qualitative research findings are used for tailoring and improving the design of the interventions for a better fit with daily primary care practice. Moreover, most qualitative findings are used for tailoring the interventions' contextual circumstances so that the interventions are performed in practice as planned, rather than adjusted to local circumstances. CONCLUSIONS: Pragmatic trials seem to be oxymoronic. Although the pragmatic trial methodology establishes the effectiveness of interventions under natural, non-experimental conditions, no pragmatic fit is allowed. Qualitative research's contribution to the development of tailor-made community-based interventions lies in providing ongoing evaluations of the dilemmas faced in pragmatic trials and allowing for the development of true tailor-made interventions. PMID- 19561173 TI - Changes in alcohol consumption and drinking patterns during 11 years of follow-up among ageing men: the FinDrink study. AB - BACKGROUND: Alcohol consumption is often reported to decrease with ageing. We investigated alcohol consumption and drinking patterns in an ageing population based male sample during an 11-year follow-up period. METHODS: This study with baseline and two follow-up examinations (at 4 and 11 years) included 1516 randomly selected participants, aged 42, 48, 54 and 60 years from Eastern Finland. Alcohol consumption and drinking patterns during the year preceding the examination were assessed. Data were analysed using Generalized Estimating Equations and Mixed Models. RESULTS: Over the 11-year study period, the amount of alcohol consumed weekly increased among the 42-year-olds (P < 0.001) and remained constant among the older cohorts. The risk of frequent drinking (alcohol consumption at least twice weekly) increased among all cohorts (OR = 2.04, 95% CI = 1.50-2.79 for 42-year-olds; OR = 1.71, 95% CI = 1.13-2.58 for 48-year-olds; OR = 1.67, 95% CI = 1.16-2.39 for 54-year-olds and OR = 1.67, 95% CI = 1.21-2.29 for 60-year-olds). There was also an increasing probability of heavy consumption (more than 14 weekly drinks) among the 42-year-olds (OR = 1.47, 95% CI = 1.09 2.00). The risk of binging (six-plus drinks at one occasion) decreased among the older participants (OR = 0.65, 95% CI = 0.47-0.89 for 54-year-olds, and OR = 0.56, 95% CI = 0.39-0.81 for 60-year-olds). CONCLUSION: Finnish men born in 1926 1946 do not seem to decrease drinking while ageing. In contrast those born in 1944-1946 increase drinking until their 60's. This should be taken into consideration in planning health services for aged men in the near future. PMID- 19561174 TI - Effects of sports injury prevention training on the biomechanical risk factors of anterior cruciate ligament injury in high school female basketball players. AB - BACKGROUND: Female athletes have a higher risk of anterior cruciate ligament injury than their male counterparts who play at similar levels in sports involving pivoting and landing. HYPOTHESIS: The competitive female basketball players who participated in a sports injury prevention training program would show better muscle strength and flexibility and improved biomechanical properties associated with anterior cruciate ligament injury than during the pretraining period and than posttraining parameters in a control group. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 22 high school female basketball players were recruited and randomly divided into 2 groups (the experimental group and the control group, 11 participants each). The experimental group was instructed in the 6 parts of the sports injury prevention training program and performed it during the first 20 minutes of team practice for the next 8 weeks, while the control group performed their regular training program. Both groups were tested with a rebound-jump task before and after the 8-week period. A total of 21 reflective markers were placed in preassigned positions. In this controlled laboratory study, a 2-way analysis of variance (2 x 2) experimental design was used for the statistical analysis (P < .05) using the experimental group and a testing session as within and between factors, respectively. Post hoc tests with Sidak correction were used when significant factor effects and/or interactions were observed. RESULTS: A comparison of the experimental group's pretraining and posttraining results identified training effects on all strength parameters (P = .004 to .043) and on knee flexion, which reflects increased flexibility (P = .022). The experimental group showed higher knee flexion angles (P = .024), greater interknee distances (P = .004), lower hamstring-quadriceps ratios (P = .023), and lower maximum knee extension torques (P = .043) after training. In the control group, no statistical differences were observed between pretraining and posttraining findings (P = .084 to .873). At pretraining, no significant differences were observed between the 2 groups for any parameter (P = .067 to .784). However, a comparison of the 2 groups after training revealed that the experimental group had significantly higher knee flexion angles (P = .023), greater knee distances (P = .005), lower hamstring-quadriceps ratios (P = .021), lower maximum knee extension torques (P = .124), and higher maximum knee abduction torques P (= .043) than the control group. CONCLUSION: The sports injury prevention training program improved the strength and flexibility of the competitive female basketball players tested and biomechanical properties associated with anterior cruciate ligament injury as compared with pretraining parameters and with posttraining parameters in the control group. CLINICAL RELEVANCE: This injury prevention program could potentially modify the flexibility, strength, and biomechanical properties associated with ACL injury and lower the athlete's risk for injury. PMID- 19561175 TI - Current concepts in the diagnosis and treatment of osteochondral lesions of the ankle. AB - Osteochondral lesions of the ankle are a more common source of ankle pain than previously recognized. Although the exact pathophysiology of the condition has not been clearly established, it is likely that a variety of etiological factors play a role, with trauma, typically from ankle sprains, being the most common. Technological advancements in ankle arthroscopy and radiologic imaging, most importantly magnetic resonance imaging, have improved diagnostic capabilities for detecting osteochondral lesions of the ankle. Moreover, these technologies have allowed for the development of more sophisticated classification systems that may, in due course, direct specific future treatment strategies. Nonoperative treatment yields best results when employed in select pediatric and adolescent patients with osteochondritis dissecans. However, operative treatment, which is dependent on the size and site of the lesion, as well as the presence or absence of cartilage damage, is frequently warranted in both children and adults with osteochondral lesions. Arthroscopic microdrilling, micropicking, and open procedures, such as osteochondral autograft transfer system and matrix-induced autologous chondrocyte implantation, are frequently employed. The purpose of this article is to review the history, etiology, and classification systems for osteochondral lesions of the ankle, as well as to describe current approaches to diagnosis and management. PMID- 19561176 TI - Comparison of landing biomechanics between male and female professional dancers. AB - BACKGROUND: The incidence of anterior cruciate ligament injuries among dancers is much lower than that among team sport athletes and no clear gender disparity has been reported in the dance population. Although numerous studies have observed differences in lower extremity landing biomechanics between male and female athletes, there is currently little research examining the landing biomechanics of male and female dancers. Comparing landing biomechanics within this population may help explain the lower overall anterior cruciate ligament injury rates and the lack of gender disparity. HYPOTHESIS: Due to the fact that dancers receive jump-specific and balance-specific training from a very young age, we hypothesized that there would be no gender differences in drop-landing biomechanics in professional dancers. STUDY DESIGN: Controlled laboratory study. METHODS: Kinematics and ground-reaction forces were recorded as 33 professional modern and ballet dancers (12 men and 21 women) performed single-legged drop landings from a 30-cm platform. Joint kinematics and kinetics were compared between genders. RESULTS: No gender differences in joint kinematics or kinetics were found during landings (multivariate analysis of variance: P = .490 and P = .175, respectively). A significant relationship was found between the age at which the dancers began training and the peak hip adduction angle during landing (r = .358, P = .041). CONCLUSION: In executing a 30-cm drop landing, male and female dancers exhibited similar landing strategies and avoided landing patterns previously associated with increased injury rates. CLINICAL RELEVANCE: Commonly reported biomechanical differences between men and women, as well as the gender disparity among athletes in the incidence of ACL injuries, may be the result of inadequate experience in proper balance and landing technique rather than intrinsic gender factors. Beginning jump-specific and balance-specific training at an early age may counteract the potentially harmful adaptations in landing biomechanics observed in female athletes after maturity. PMID- 19561177 TI - Metabolic capacity of Sinorhizobium (Ensifer) meliloti strains as determined by phenotype MicroArray analysis. AB - Sinorhizobium meliloti is a soil bacterium that fixes atmospheric nitrogen in plant roots. The high genetic diversity of its natural populations has been the subject of extensive analysis. Recent genomic studies of several isolates revealed a high content of variable genes, suggesting a correspondingly large phenotypic differentiation among strains of S. meliloti. Here, using the Phenotype MicroArray (PM) system, hundreds of different growth conditions were tested in order to compare the metabolic capabilities of the laboratory reference strain Rm1021 with those of four natural S. meliloti isolates previously analyzed by comparative genomic hybridization (CGH). The results of PM analysis showed that most phenotypic differences involved carbon source utilization and tolerance to osmolytes and pH, while fewer differences were scored for nitrogen, phosphorus, and sulfur source utilization. Only the variability of the tested strain in tolerance to sodium nitrite and ammonium sulfate of pH 8 was hypothesized to be associated with the genetic polymorphisms detected by CGH analysis. Colony and cell morphologies and the ability to nodulate Medicago truncatula plants were also compared, revealing further phenotypic diversity. Overall, our results suggest that the study of functional (phenotypic) variability of S. meliloti populations is an important and complementary step in the investigation of genetic polymorphism of rhizobia and may help to elucidate rhizobial evolutionary dynamics, including adaptation to diverse environments. PMID- 19561179 TI - Monochloramine disinfection kinetics of Nitrosomonas europaea by propidium monoazide quantitative PCR and Live/dead BacLight methods. AB - Monochloramine disinfection kinetics were determined for the pure-culture ammonia oxidizing bacterium Nitrosomonas europaea (ATCC 19718) by two culture-independent methods, namely, Live/Dead BacLight (LD) and propidium monoazide quantitative PCR (PMA-qPCR). Both methods were first verified with mixtures of heat-killed (nonviable) and non-heat-killed (viable) cells before a series of batch disinfection experiments with stationary-phase cultures (batch grown for 7 days) at pH 8.0, 25 degrees C, and 5, 10, and 20 mg Cl(2)/liter monochloramine. Two data sets were generated based on the viability method used, either (i) LD or (ii) PMA-qPCR. These two data sets were used to estimate kinetic parameters for the delayed Chick-Watson disinfection model through a Bayesian analysis implemented in WinBUGS. This analysis provided parameter estimates of 490 mg Cl(2)-min/liter for the lag coefficient (b) and 1.6 x 10(-3) to 4.0 x 10(-3) liter/mg Cl(2)-min for the Chick-Watson disinfection rate constant (k). While estimates of b were similar for both data sets, the LD data set resulted in a greater k estimate than that obtained with the PMA-qPCR data set, implying that the PMA-qPCR viability measure was more conservative than LD. For N. europaea, the lag phase was not previously reported for culture-independent methods and may have implications for nitrification in drinking water distribution systems. This is the first published application of a PMA-qPCR method for disinfection kinetic model parameter estimation as well as its application to N. europaea or monochloramine. Ultimately, this PMA-qPCR method will allow evaluation of monochloramine disinfection kinetics for mixed-culture bacteria in drinking water distribution systems. PMID- 19561180 TI - Sulfur isotope enrichment during maintenance metabolism in the thermophilic sulfate-reducing bacterium Desulfotomaculum putei. AB - Values of Delta(34)S (=delta(34)S(HS)-delta(34)S(SO(4)), where delta(34)S(HS) and delta(34)S(SO(4)) indicate the differences in the isotopic compositions of the HS(-) and SO(4)(2-) in the eluent, respectively) for many modern marine sediments are in the range of -55 to -75 per thousand, much greater than the -2 to -46 per thousand epsilon(34)S (kinetic isotope enrichment) values commonly observed for microbial sulfate reduction in laboratory batch culture and chemostat experiments. It has been proposed that at extremely low sulfate reduction rates under hypersulfidic conditions with a nonlimited supply of sulfate, isotopic enrichment in laboratory culture experiments should increase to the levels recorded in nature. We examined the effect of extremely low sulfate reduction rates and electron donor limitation on S isotope fractionation by culturing a thermophilic, sulfate-reducing bacterium, Desulfotomaculum putei, in a biomass recycling culture vessel, or "retentostat." The cell-specific rate of sulfate reduction and the specific growth rate decreased progressively from the exponential phase to the maintenance phase, yielding average maintenance coefficients of 10(-16) to 10(-18) mol of SO(4) cell(-1) h(-1) toward the end of the experiments. Overall S mass and isotopic balance were conserved during the experiment. The differences in the delta(34)S values of the sulfate and sulfide eluting from the retentostat were significantly larger, attaining a maximum Delta(34)S of -20.9 per thousand, than the -9.7 per thousand observed during the batch culture experiment, but differences did not attain the values observed in marine sediments. PMID- 19561181 TI - Widespread distribution of poribacteria in demospongiae. AB - Poribacteria were found in nine sponge species belonging to six orders of Porifera from three oceans. Phylogenetic analysis revealed four distinct poribacterial clades, which contained organisms obtained from several different geographic regions, indicating that the distribution of poribacteria is cosmopolitan. Members of divergent poribacterial clades were also found in the same sponge species in three different sponge genera. PMID- 19561182 TI - RNA colony blot hybridization method for enumeration of culturable Vibrio cholerae and Vibrio mimicus bacteria. AB - A species-specific RNA colony blot hybridization protocol was developed for enumeration of culturable Vibrio cholerae and Vibrio mimicus bacteria in environmental water samples. Bacterial colonies on selective or nonselective plates were lysed by sodium dodecyl sulfate, and the lysates were immobilized on nylon membranes. A fluorescently labeled oligonucleotide probe targeting a phylogenetic signature sequence of 16S rRNA of V. cholerae and V. mimicus was hybridized to rRNA molecules immobilized on the nylon colony lift blots. The protocol produced strong positive signals for all colonies of the 15 diverse V. cholerae-V. mimicus strains tested, indicating 100% sensitivity of the probe for the targeted species. For visible colonies of 10 nontarget species, the specificity of the probe was calculated to be 90% because of a weak positive signal produced by Grimontia (Vibrio) hollisae, a marine bacterium. When both the sensitivity and specificity of the assay were evaluated using lake water samples amended with a bioluminescent V. cholerae strain, no false-negative or false positive results were found, indicating 100% sensitivity and specificity for culturable bacterial populations in freshwater samples when G. hollisae was not present. When the protocol was applied to laboratory microcosms containing V. cholerae attached to live copepods, copepods were found to carry approximately 10,000 to 50,000 CFU of V. cholerae per copepod. The protocol was also used to analyze pond water samples collected in an area of cholera endemicity in Bangladesh over a 9-month period. Water samples collected from six ponds demonstrated a peak in abundance of total culturable V. cholerae bacteria 1 to 2 months prior to observed increases in pathogenic V. cholerae and in clinical cases recorded by the area health clinic. The method provides a highly specific and sensitive tool for monitoring the dynamics of V. cholerae in the environment. The RNA blot hybridization protocol can also be applied to detection of other gram-negative bacteria for taxon-specific enumeration. PMID- 19561183 TI - Dead-end hollow-fiber ultrafiltration for recovery of diverse microbes from water. AB - Dead-end ultrafiltration (DEUF) is an alternative approach to tangential-flow hollow-fiber ultrafiltration that can be readily employed under field conditions to recover microbes from water. The hydraulics of DEUF and microbe recovery for a new DEUF method were investigated using 100-liter tap water samples. Pressure, flow rate, and temperature were investigated using four hollow-fiber ultrafilter types. Based on hydraulic performance, the Asahi Kasei REXEED 25S ultrafilter was selected for microbe recovery experiments. Microbe recovery experiments were performed using MS2 bacteriophage, Enterococcus faecalis, Clostridium perfringens spores, and Cryptosporidium parvum oocysts. Microbes were recovered from ultrafilters by backflushing using a surfactant solution. Average flow rates were 2.1 liters/min for 100-liter water samples having turbidities of 0.28 to 4.3 nephelometric turbidity units (NTU), and no evidence of appreciable filter clogging was observed. The DEUF average recovery efficiencies for each study analyte in tap water were as follows: for E. faecalis, 93% +/- 16%; for MS2, 57% +/- 7.7%; for C. perfringens spores, 94% +/- 22%; and for C. parvum, 87% +/- 18%. Average microbe recoveries for tap water amended with surface water (average turbidity = 4.3 NTU) were as follows: for E. faecalis, 78% +/- 12%; for MS2, 73% +/- 13%; for C. perfringens, 57% +/- 21%; and for C. parvum, 83% +/- 21%. These data demonstrate that DEUF is an effective method for recovering diverse microbes from water and should be a useful tool for field-based environmental investigations. PMID- 19561184 TI - Identification of a novel two-peptide lantibiotic, lichenicidin, following rational genome mining for LanM proteins. AB - Lantibiotics are ribosomally synthesized peptide antimicrobials which contain considerable posttranslational modifications. Given their usually broad host range and their highly stable structures, there have been renewed attempts to identify and characterize novel members of the lantibiotic family in recent years. The increasing availability of bacterial genome sequences means that in addition to traditional microbiological approaches, in silico screening strategies may now be employed to the same end. Taking advantage of the highly conserved nature of lantibiotic biosynthetic enzymes, we screened publicly available microbial genome sequences for genes encoding LanM proteins, which are required for the posttranslational modification of type 2 lantibiotics. By using this approach, 89 LanM homologs, including 61 in strains not known to be lantibiotic producers, were identified. Of these strains, five (Streptococcus pneumoniae SP23-BS72, Bacillus licheniformis ATCC 14580, Anabaena variabilis ATCC 29413, Geobacillus thermodenitrificans NG80-2, and Herpetosiphon aurantiacus ATCC 23779) were subjected to a more detailed bioinformatic analysis. Four of the strains possessed genes potentially encoding a structural peptide in close proximity to the lanM determinants, while two, S. pneumoniae SP23-BS72 and B. licheniformis ATCC 14580, possess two potential structural genes. The B. licheniformis strain was selected for a proof-of-concept exercise, which established that a two-peptide lantibiotic, lichenicidin, which exhibits antimicrobial activity against all Listeria monocytogenes, methicillin-resistant Staphylococcus aureus, and vancomycin-resistant enterococcus strains tested, was indeed produced, thereby confirming the benefits of such a bioinformatic approach when screening for novel lantibiotic producers. PMID- 19561185 TI - Rhodococcus rhodochrous DSM 43269 3-ketosteroid 9alpha-hydroxylase, a two component iron-sulfur-containing monooxygenase with subtle steroid substrate specificity. AB - This paper reports the biochemical characterization of a purified and reconstituted two-component 3-ketosteroid 9alpha-hydroxylase (KSH). KSH of Rhodococcus rhodochrous DSM 43269, consisting of a ferredoxin reductase (KshB) and a terminal oxygenase (KshA), was heterologously expressed in Escherichia coli. E. coli cell cultures, expressing both KshA and KshB, converted 4 androstene-3,17-dione (AD) into 9alpha-hydroxy-4-AD (9OHAD) with a >60% molar yield over 48 h of incubation. Coexpression and copurification were critical to successfully obtain pure and active KSH. Biochemical analysis revealed that the flavoprotein KshB is an NADH-dependent reductase using flavin adenine dinucleotide as a cofactor. Reconstitution experiments confirmed that KshA, KshB, and NADH are essential for KSH activity with steroid substrates. KSH hydroxylation activity was inhibited by several divalent metal ions, especially by zinc. The reconstituted KSH displayed subtle steroid substrate specificity; a range of 3-ketosteroids, i.e., 5alpha-Eta, 5beta-Eta, Delta1, and Delta4 steroids, could act as KSH substrates, provided that they had a short side chain. The formation of 9OHAD from AD by KSH was confirmed by liquid chromatography-mass spectrometry analysis and by the specific enzymatic conversion of 9OHAD into 3 hydroxy-9,10-secoandrost-1,3,5(10)-triene-9,17-dione using 3-ketosteroid Delta1 dehydrogenase. Only a single KSH is encoded in the genome of the human pathogen Mycobacterium tuberculosis H37Rv, shown to be important for survival in macrophages. Since no human KSH homolog exists, the M. tuberculosis enzyme may provide a novel target for treatment of tuberculosis. Detailed knowledge about the biochemical properties of KSH thus is highly relevant in the research fields of biotechnology and medicine. PMID- 19561178 TI - Comparison of species richness estimates obtained using nearly complete fragments and simulated pyrosequencing-generated fragments in 16S rRNA gene-based environmental surveys. AB - Pyrosequencing-based 16S rRNA gene surveys are increasingly utilized to study highly diverse bacterial communities, with special emphasis on utilizing the large number of sequences obtained (tens to hundreds of thousands) for species richness estimation. However, it is not yet clear how the number of operational taxonomic units (OTUs) and, hence, species richness estimates determined using shorter fragments at different taxonomic cutoffs correlates with the number of OTUs assigned using longer, nearly complete 16S rRNA gene fragments. We constructed a 16S rRNA clone library from an undisturbed tallgrass prairie soil (1,132 clones) and used it to compare species richness estimates obtained using eight pyrosequencing candidate fragments (99 to 361 bp in length) and the nearly full-length fragment. Fragments encompassing the V1 and V2 (V1+V2) region and the V6 region (generated using primer pairs 8F-338R and 967F-1046R) overestimated species richness; fragments encompassing the V3, V7, and V7+V8 hypervariable regions (generated using primer pairs 338F-530R, 1046F-1220R, and 1046F-1392R) underestimated species richness; and fragments encompassing the V4, V5+V6, and V6+V7 regions (generated using primer pairs 530F-805R, 805F-1046R, and 967F 1220R) provided estimates comparable to those obtained with the nearly full length fragment. These patterns were observed regardless of the alignment method utilized or the parameter used to gauge comparative levels of species richness (number of OTUs observed, slope of scatter plots of pairwise distance values for short and nearly complete fragments, and nonparametric and parametric species richness estimates). Similar results were obtained when analyzing three other datasets derived from soil, adult Zebrafish gut, and basaltic formations in the East Pacific Rise. Regression analysis indicated that these observed discrepancies in species richness estimates within various regions could readily be explained by the proportions of hypervariable, variable, and conserved base pairs within an examined fragment. PMID- 19561186 TI - Culture-independent characterization of archaeal biodiversity in swine confinement building bioaerosols. AB - It was previously demonstrated that microbial communities of pig manure were composed of both bacteria and archaea. Recent studies have shown that bacteria are aerosolized from pig manure, but none have ever focused on the airborne archaeal burden. We sought here to develop and apply molecular ecology approaches to thoroughly characterize airborne archaea from swine confinement buildings (SCBs). Eight swine operations were visited, twice in winter and once during summer. Institute of Occupational Medicine cassettes loaded with 25-mm gelatin filters were used to capture the inhalable microbial biomass. The total genomic DNA was extracted and used as a template for PCR amplification of the archaeal 16S rRNA gene. High concentrations of archaea were found in SCB bioaerosols, being as high as 10(8) 16S rRNA gene copies per cubic meter of air. Construction and sequencing of 16S rRNA gene libraries revealed that all sequences were closely related to methanogenic archaea, such as Methanosphaera stadtmanae (94.7% of the archaeal biodiversity). Archaeal community profiles were compared by 16S rRNA gene denaturing gradient gel electrophoresis. This analysis showed similar fingerprints in each SCB and confirmed the predominance of methanogenic archaea in the bioaerosols. This study sheds new light on the nature of bioaerosols in SCBs and suggests that archaea are also aerosolized from pig manure. PMID- 19561187 TI - Adsorption of Mycobacterium avium subsp. paratuberculosis to soil particles. AB - Attachment of Mycobacterium avium subsp. paratuberculosis to soil particles could increase their availability to farm animals, as well as influence the transportation of M. avium subsp. paratuberculosis to water sources. To investigate the possibility of such attachment, we passed a known quantity of M. avium subsp. paratuberculosis through chromatography columns packed with clay soil, sandy soil, pure silica, clay-silica mixture, or clay-silica complexes and measured the organisms recovered in the eluent using culture or quantitative PCR. Experiments were repeated using buffer at a range of pH levels with pure silica to investigate the effect of pH on M. avium subsp. paratuberculosis attachment. Linear mixed-model analyses were conducted to compare the proportional recovery of M. avium subsp. paratuberculosis in the eluent between different substrates and pH levels. Of the organisms added to the columns, 83 to 100% were estimated to be retained in the columns after adjustment for those retained in empty control columns. The proportions recovered were significantly different across different substrates, with the retention being significantly greater (P < 0.05) in pure substrates (silica and clay-silica complexes) than in soil substrates (clay soil and sandy soil). However, there were no significant differences in the retention of M. avium subsp. paratuberculosis between silica and clay-silica complexes or between clay soil and sandy soil. The proportion retained decreased with increasing pH in one of the experiments, indicating greater adsorption of M. avium subsp. paratuberculosis to soil particles at an acidic pH (P < 0.05). The results suggest that under experimental conditions M. avium subsp. paratuberculosis adsorbs to a range of soil particles, and this attachment is influenced by soil pH. PMID- 19561188 TI - Development of a cell culture method to isolate and enrich Salmonella enterica serotype enteritidis from shell eggs for subsequent detection by real-time PCR. AB - Salmonella enterica serotype Enteritidis is a major cause of nontyphoidal salmonellosis from ingestion of contaminated raw or undercooked shell eggs. Current techniques used to identify Salmonella serotype Enteritidis in eggs are extremely laborious and time-consuming. In this study, a novel eukaryotic cell culture system was combined with real-time PCR analysis to rapidly identify Salmonella serotype Enteritidis in raw shell eggs. The system was compared to the standard microbiological method of the International Organization for Standardization (Anonymous, Microbiology of food and animal feeding stuffs horizontal method for the detection of Salmonella, 2002). The novel technique utilizes a mouse macrophage cell line (RAW 264.7) as the host for the isolation and intracellular replication of Salmonella serotype Enteritidis. Exposure of macrophages to Salmonella serotype Enteritidis-contaminated eggs results in uptake and intracellular replication of the bacterium, which can subsequently be detected by real-time PCR analysis of the DNA released after disruption of infected macrophages. Macrophage monolayers were exposed to eggs contaminated with various quantities of Salmonella serotype Enteritidis. As few as 10 CFU/ml was detected in cell lysates from infected macrophages after 10 h by real-time PCR using primer and probe sets specific for DNA segments located on the Salmonella serotype Enteritidis genes sefA and orgC. Salmonella serotype Enteritidis could also be distinguished from other non-serogroup D Salmonella serotypes by using the sefA- and orgC-specific primer and probe sets. Confirmatory identification of Salmonella serotype Enteritidis in eggs was also achieved by isolation of intracellular bacteria from lysates of infected macrophages on xylose lysine deoxycholate medium. This method identifies Salmonella serotype Enteritidis from eggs in less than 10 h compared to the more than 5 days required for the standard reference microbiological method of the International Organization for Standardization (Microbiology of food and animal feeding stuffs-horizontal method for the detection of Salmonella, 2002). PMID- 19561190 TI - Identification and characterization of a novel intracellular poly(3 hydroxybutyrate) depolymerase from Bacillus megaterium. AB - A gene that codes for a novel intracellular poly(3-hydroxybutyrate) (PHB) depolymerase, designated PhaZ1, has been identified in the genome of Bacillus megaterium. A native PHB (nPHB) granule-binding assay showed that purified soluble PhaZ1 had strong affinity for nPHB granules. Turbidimetric analyses revealed that PhaZ1 could rapidly degrade nPHB granules in vitro without the need for protease pretreatment of the granules to remove surface proteins. Notably, almost all the final hydrolytic products produced from the in vitro degradation of nPHB granules by PhaZ1 were 3-hydroxybutyric acid (3HB) monomers. Unexpectedly, PhaZ1 could also hydrolyze denatured semicrystalline PHB, with the generation of 3HB monomers. The disruption of the phaZ1 gene significantly affected intracellular PHB mobilization during the PHB-degrading stage in B. megaterium, as demonstrated by transmission electron microscopy and the measurement of the PHB content. These results indicate that PhaZ1 is functional in intracellular PHB mobilization in vivo. Some of these features, which are in striking contrast with those of other known nPHB granule-degrading PhaZs, may provide an advantage for B. megaterium PhaZ1 in fermentative production of the biotechnologically valuable chiral compound (R)-3HB. PMID- 19561191 TI - Evaluation of different methods for extracting extracellular DNA from the biofilm matrix. AB - The occurrence of high concentrations of extracellular DNA (eDNA) in the extracellular matrices of biofilms plays an important role in biofilm formation and development and possibly in horizontal gene transfer through natural transformation. Studies have been conducted to characterize the nature of eDNA and its potential function in biofilm development, but it is difficult to extract eDNA from the extracellular matrices of biofilms without any contamination from genomic DNA released by cell lysis during the extraction process. In this report, we compared several different extraction methods in order to obtain highly pure eDNA from different biofilm samples. After different extraction methods were explored, it was concluded that using chemical treatment or enzymatic treatment of biofilm samples may obtain larger amounts of eDNA than using the simple filtration method. There was no detectable cell lysis when the enzymatic treatment methods were used, but substantial cell lysis was observed when the chemical treatment methods were used. These data suggest that eDNA may bind to other extracellular polymers in the biofilm matrix and that enzymatic treatment methods are effective and favorable for extracting eDNA from biofilm samples. Moreover, randomly amplified polymorphic DNA analysis of eDNA in Acinetobacter sp. biofilms and Acinetobacter sp. genomic DNA and DNA sequencing analysis revealed that eDNA originated from genomic DNA but was not structurally identical to the genomic DNA. PMID- 19561192 TI - Relative diversity and community structure of ciliates in stream biofilms according to molecular and microscopy methods. AB - Ciliates are an important component of aquatic ecosystems, acting as predators of bacteria and protozoa and providing nutrition for organisms at higher trophic levels. Understanding of the diversity and ecological role of ciliates in stream biofilms is limited, however. Ciliate diversity in biofilm samples from four streams subject to different impacts by human activity was assessed using microscopy and terminal restriction fragment length polymorphism (T-RFLP) analysis of 18S rRNA sequences. Analysis of 3' and 5' terminal fragments yielded very similar estimates of ciliate diversity. The diversity detected using microscopy was consistently lower than that suggested by T-RFLP analysis, indicating the existence of genetic diversity not apparent by morphological examination. Microscopy and T-RFLP analyses revealed similar relative trends in diversity between different streams, with the lowest level of biofilm-associated ciliate diversity found in samples from the least-impacted stream and the highest diversity in samples from moderately to highly impacted streams. Multivariate analysis provided evidence of significantly different ciliate communities in biofilm samples from different streams and seasons, particularly between a highly degraded urban stream and less impacted streams. Microscopy and T-RFLP data both suggested the existence of widely distributed, resilient biofilm-associated ciliates as well as ciliate taxa restricted to sites with particular environmental conditions, with cosmopolitan taxa being more abundant than those with restricted distributions. Differences between ciliate assemblages were associated with water quality characteristics typical of urban stream degradation and may be related to factors such as nutrient availability and macroinvertebrate communities. Microscopic and molecular techniques were considered to be useful complementary approaches for investigation of biofilm ciliate communities. PMID- 19561193 TI - Position of eukaryotic translation initiation factor eIF1A on the 40S ribosomal subunit mapped by directed hydroxyl radical probing. AB - The universally conserved eukaryotic initiation factor (eIF), eIF1A, plays multiple roles throughout initiation: it stimulates eIF2/GTP/Met-tRNA(i)(Met) attachment to 40S ribosomal subunits, scanning, start codon selection and subunit joining. Its bacterial ortholog IF1 consists of an oligonucleotide/oligosaccharide-binding (OB) domain, whereas eIF1A additionally contains a helical subdomain, N-terminal tail (NTT) and C-terminal tail (CTT). The NTT and CTT both enhance ribosomal recruitment of eIF2/GTP/Met-tRNA(i)(Met), but have opposite effects on the stringency of start codon selection: the CTT increases, whereas the NTT decreases it. Here, we determined the position of eIF1A on the 40S subunit by directed hydroxyl radical cleavage. eIF1A's OB domain binds in the A site, similar to IF1, whereas the helical subdomain contacts the head, forming a bridge over the mRNA channel. The NTT and CTT both thread under Met-tRNA(i)(Met) reaching into the P-site. The NTT threads closer to the mRNA channel. In the proposed model, the NTT does not clash with either mRNA or Met tRNA(i)(Met), consistent with its suggested role in promoting the 'closed' conformation of ribosomal complexes upon start codon recognition. In contrast, eIF1A-CTT appears to interfere with the P-site tRNA-head interaction in the 'closed' complex and is likely ejected from the P-site upon start codon recognition. PMID- 19561194 TI - JunB mediates enhancer/promoter activity of COL1A2 following TGF-beta induction. AB - Transcriptional control of the genes coding for collagen type I is regulated by a complex interaction between a distal enhancer and a proximal promoter. In this study, we have dissected the molecular mechanism of this interaction by defining a specific sequence within the enhancer that respond in fibroblasts to transforming growth factor-beta (TGF-beta). We show that TGF-beta activates COL1A2 gene via a non-canonical (Smad-independent) signalling pathway, which requires enhancer/promoter co-operation. This interaction involves exchange of cJun/Jun B transcription factor occupancy of a critical enhancer site resulting in the stabilization of enhancer/promoter coalescence. Moreover, using transgenesis, we show that interference in this mechanism results in the abolition of COL1A2 fibroblast expression in vivo. These data are therefore relevant to the control of collagen type I in vivo both in embryonic development, in adult connective tissue homeostasis, and in tissue repair and scarring pathologies. PMID- 19561189 TI - Assessment of bias associated with incomplete extraction of microbial DNA from soil. AB - DNA extraction bias is a frequently cited but poorly understood limitation of molecular characterizations of environmental microbial communities. To assess the bias of a commonly used soil DNA extraction kit, we varied the cell lysis protocol and conducted multiple extractions on subsamples of clay, sand, and organic soils. DNA, as well as bacterial and fungal ribosomal gene copies as measured by quantitative PCR, continued to be isolated in successive extractions. When terminal restriction fragment length polymorphism was used, a significant shift in community composition due to extraction bias was detected for bacteria but not for fungi. Pyrosequencing indicated that the relative abundances of sequences from rarely cultivated groups such as Acidobacteria, Gemmatimonades, and Verrucomicrobia were higher in the first extraction than in the sixth but that the reverse was true for Proteobacteria and Actinobacteria. This suggests that the well-known phylum-level bacterial cultivation bias may be partially exaggerated by DNA extraction bias. We conclude that bias can be adequately reduced in many situations by pooling three successive extractions, and additional measures should be considered when divergent soil types are compared or when comprehensive community analysis is necessary. PMID- 19561195 TI - A FRET-based assay for characterization of alternative splicing events using peptide nucleic acid fluorescence in situ hybridization. AB - We describe a quantitative method for detecting RNA alternative splicing variants that combines in situ hybridization of fluorescently labeled peptide nucleic acid (PNA) probes with confocal microscopy Forster resonance energy transfer (FRET). The use of PNA probes complementary to sequences flanking a given splice junction allows to specifically quantify, within the cell, the RNA isoform generating such splice junction by FRET measure. As a proof of concept we analyzed two alternative splicing events originating from lymphocyte antigen 6 (LY6) complex, locus G5B (LY6G5B) pre-mRNA. These are characterized by the removal of the first intron (Fully Spliced Isoform, FSI) or by retention of such intron (Intron Retained Isoform, IRI). The use of PNA probe pairs labeled with donor (Cy3) and acceptor (Cy5) fluorophores, suitable to FRET, flanking FSI and IRI specific splice junctions specifically detected both mRNA isoforms in HeLa cells. We have observed that the method works efficiently with probes 5-11 nt apart. The data supports that this FRET-based PNA fluorescence in situ hybridization (FP-FISH) method offers a conceptually new approach for characterizing at the subcellular level not only splice variant isoform structure, location and dynamics but also potentially a wide variety of close range RNA-RNA interactions. PMID- 19561197 TI - Discovering ligands for a microRNA precursor with peptoid microarrays. AB - We have screened peptoid microarrays to identify specific ligands for the RNA hairpin precursor of miR-21, a microRNA involved in cancer and heart disease. Microarrays were printed by spotting a library of 7680 N-substituted oligoglycines (peptoids) onto glass slides. Two compounds on the array specifically bind RNA having the sequence and predicted secondary structure of the miR-21 precursor hairpin and have specific affinity for the target in solution. Their binding induces a conformational change around the hairpin loop, and the most specific compound recognizes the loop sequence and a bulged uridine in the proximal duplex. Functional groups contributing affinity and specificity were identified, and by varying a critical methylpyridine group, a compound with a dissociation constant of 1.9 microM for the miR-21 precursor hairpin and a 20 fold discrimination against a closely-related hairpin was created. This work describes a systematic approach to discovery of ligands for specific pre-defined novel RNA structures. It demonstrates discovery of new ligands for an RNA for which no specific lead compounds were previously known by screening a microarray of small molecules. PMID- 19561196 TI - Senescence delay and repression of p16INK4a by Lsh via recruitment of histone deacetylases in human diploid fibroblasts. AB - Lymphoid specific helicase (Lsh) belongs to the family of SNF2/helicases. Disruption of Lsh leads to developmental growth retardation and premature aging in mice. However, the specific effect of Lsh on human cellular senescence remains unknown. Herein, we report that Lsh overexpression delays cell senescence by silencing p16(INK4a) in human fibroblasts. The patterns of p16(INK4a) and Lsh expression during cell senescence present the inverse correlation. We also find that Lsh requires histone deacetylase (HDAC) activity to repress p16(INK4a) and treatment with trichostatin A (TSA) is sufficient to block the repressor effect of Lsh. Moreover, overexpression of Lsh is correlated with deacetylation of histone H3 at the p16 promoter, and TSA treatment in Lsh-expressing cells reverses the acetylation status of histones. Additionally, we demonstrate an interaction between Lsh, histone deacetylase 1 (HDAC1) and HDAC2 in vivo. Furthermore, we demonstrate that Lsh interacts in vivo with the p16 promoter and recruits HDAC1. Our data suggest that Lsh represses endogenous p16(INK4a) expression by recruiting HDAC to establish a repressive chromatin structure at the p16(INK4a) promoter, which in turn delays cell senescence. PMID- 19561198 TI - Fibroblast growth factor 1 induced during myogenesis by a transcription translation coupling mechanism. AB - Fibroblast growth factor 1 (FGF1) is involved in muscle development and regeneration. The FGF1 gene contains four tissue-specific promoters allowing synthesis of four transcripts with distinct leader regions. Two of these transcripts contain internal ribosome entry sites (IRESs), which are RNA elements allowing mRNA translation to occur in conditions of blockade of the classical cap dependent mechanism. Here, we investigated the function and the regulation of FGF1 during muscle differentiation and regeneration. Our data show that FGF1 protein expression is induced in differentiating myoblasts and regenerating mouse muscle, whereas siRNA knock-down demonstrated FGF1 requirement for myoblast differentiation. FGF1 induction occurred at both transcriptional and translational levels, involving specific activation of both promoter A and IRES A, whereas global cap-dependent translation was inhibited. Furthermore, we identified, in the FGF1 promoter A distal region, a cis-acting element able to activate the IRES A-driven translation. These data revealed a mechanism of molecular coupling of mRNA transcription and translation, involving a unique process of IRES activation by a promoter element. The crucial role of FGF1 in myoblast differentiation provides physiological relevance to this novel mechanism. This finding also provides a new insight into the molecular mechanisms linking different levels of gene expression regulation. PMID- 19561199 TI - Structural characterization of cationic lipid-tRNA complexes. AB - Despite considerable interest and investigations on cationic lipid-DNA complexes, reports on lipid-RNA interaction are very limited. In contrast to lipid-DNA complexes where lipid binding induces partial B to A and B to C conformational changes, lipid-tRNA complexation preserves tRNA folded state. This study is the first attempt to investigate the binding of cationic lipid with transfer RNA and the effect of lipid complexation on tRNA aggregation and condensation. We examine the interaction of tRNA with cholesterol (Chol), 1,2-dioleoyl-3-trimethylammonium propane (DOTAP), dioctadecyldimethylammoniumbromide (DDAB) and dioleoylphosphatidylethanolamine (DOPE), at physiological condition, using constant tRNA concentration and various lipid contents. FTIR, UV-visible, CD spectroscopic methods and atomic force microscopy (AFM) were used to analyze lipid binding site, the binding constant and the effects of lipid interaction on tRNA stability, conformation and condensation. Structural analysis showed lipid tRNA interactions with G-C and A-U base pairs as well as the backbone phosphate group with overall binding constants of K(Chol) = 5.94 (+/- 0.8) x 10(4) M(-1), K(DDAB) = 8.33 (+/- 0.90) x 10(5) M(-1), K(DOTAP) = 1.05 (+/- 0.30) x 10(5) M(-1) and K(DOPE) = 2.75 (+/- 0.50) x 10(4) M(-1). The order of stability of lipid-tRNA complexation is DDAB > DOTAP > Chol > DOPE. Hydrophobic interactions between lipid aliphatic tails and tRNA were observed. RNA remains in A-family structure, while biopolymer aggregation and condensation occurred at high lipid concentrations. PMID- 19561200 TI - Conserved principles of mammalian transcriptional regulation revealed by RNA half life. AB - RNA levels in a cell are regulated by the relative rates of RNA synthesis and decay. We recently developed a new approach for measuring both RNA synthesis and decay in a single experimental setting by biosynthetic labeling of newly transcribed RNA. Here, we show that this provides measurements of RNA half-lives from microarray data with a so far unreached accuracy. Based on such measurements of RNA half-lives for human B-cells and mouse fibroblasts, we identified conserved regulatory principles for a large number of biological processes. We show that different regulatory patterns between functionally similar proteins are characterized by differences in the half-life of the corresponding transcripts and can be identified by measuring RNA half-life. We identify more than 100 protein families which show such differential regulatory patterns in both species. Additionally, we provide strong evidence that the activity of protein complexes consisting of subunits with overall long transcript half-lives can be regulated by transcriptional regulation of individual key subunits with short lived transcripts. Based on this observation, we predict more than 100 key regulatory subunits for human complexes of which 28% could be confirmed in mice (P < 10(-9)). Therefore, this atlas of transcript half-lives provides new fundamental insights into many cellular processes. PMID- 19561201 TI - One small leap for the jumping spider but a giant step for vision science. PMID- 19561202 TI - Do cephalopods communicate using polarized light reflections from their skin? AB - Cephalopods (squid, cuttlefish and octopus) are probably best known for their ability to change color and pattern for camouflage and communication. This is made possible by their complex skin, which contains pigmented chromatophore organs and structural light reflectors (iridophores and leucophores). Iridophores create colorful and linearly polarized reflective patterns. Equally interesting, the photoreceptors of cephalopod eyes are arranged in a way to give these animals the ability to detect the linear polarization of incoming light. The capacity to detect polarized light may have a variety of functions, such as prey detection, navigation, orientation and contrast enhancement. Because the skin of cephalopods can produce polarized reflective patterns, it has been postulated that cephalopods could communicate intraspecifically through this visual system. The term 'hidden' or 'private' communication channel has been given to this concept because many cephalopod predators may not be able to see their polarized reflective patterns. We review the evidence for polarization vision as well as polarization signaling in some cephalopod species and provide examples that tend to support the notion--currently unproven--that some cephalopods communicate using polarized light signals. PMID- 19561203 TI - Naive bats discriminate arctiid moth warning sounds but generalize their aposematic meaning. AB - Naive red (Lasiurus borealis Muller) and big brown (Eptesicus fuscus Beauvois) bats quickly learn to avoid noxious sound-producing tiger moths. After this experience with a model tiger moth, bats generalize the meaning of these prey generated sounds to a second tiger moth species producing a different call. Here we describe the three-dimensional kinematic and bioacoustic details of this behaviour, first, as naive bats learn to deal with an unpalatable model tiger moth and subsequently, as they avoid acoustic mimics. The tiger moths' first clicks influenced the bats' echolocation behaviour and the percentage of interactions that included terminal buzzes was associated with capture and investigatory behaviour. When the mimic was introduced, the bats decreased both their minimum distance to the tiger moth and the time at which they broke off their attack compared with their exposure to the model on the night before. These kinematic signatures closely match the bats' behaviour on their first night of experience with the model. Minimum distances and time of pursuit cessation increased again by the last night of the mimic's presentation. These kinematic and bioacoustic results show that although naive bats generalize the meaning of aposematic tiger moth calls, they discriminate the prey-generated signals as different and investigate. Extrapolating to experienced bats, these results suggest that acoustic predators probably exert potent and fine-scaled selective forces on acoustic mimicry complexes. PMID- 19561204 TI - Lift and drag performance of odontocete cetacean flippers. AB - Cetaceans (whales, dolphins and porpoises) have evolved flippers that aid in effective locomotion through their aquatic environments. Differing evolutionary pressures upon cetaceans, including hunting and feeding requirements, and other factors such as animal mass and size have resulted in flippers that are unique among each species. Cetacean flippers may be viewed as being analogous to modern engineered hydrofoils, which have hydrodynamic properties such as lift coefficient, drag coefficient and associated efficiency. Field observations and the collection of biological samples have resulted in flipper geometry being known for most cetacean species. However, the hydrodynamic properties of cetacean flippers have not been rigorously examined and thus their performance properties are unknown. By conducting water tunnel testing using scale models of cetacean flippers derived via computed tomography (CT) scans, as well as computational fluid dynamic (CFD) simulations, we present a baseline work to describe the hydrodynamic properties of several cetacean flippers. We found that flippers of similar planform shape had similar hydrodynamic performance properties. Furthermore, one group of flippers of planform shape similar to modern swept wings was found to have lift coefficients that increased with angle of attack nonlinearly, which was caused by the onset of vortex-dominated lift. Drag coefficient versus angle of attack curves were found to be less dependent on planform shape. Our work represents a step towards the understanding of the association between performance, ecology, morphology and fluid mechanics based on the three-dimensional geometry of cetacean flippers. PMID- 19561205 TI - Post-prandial metabolic alkalosis in the seawater-acclimated trout: the alkaline tide comes in. AB - The consequences of feeding and digestion on acid-base balance and regulation in a marine teleost (seawater-acclimated steelhead trout; Oncorhynchus mykiss) were investigated by tracking changes in blood pH and [HCO3-], as well as alterations in net acid or base excretion to the water following feeding. Additionally the role of the intestine in the regulation of acid-base balance during feeding was investigated with an in vitro gut sac technique. Feeding did not affect plasma glucose or urea concentrations, however, total plasma ammonia rose during feeding, peaking between 3 and 24 h following the ingestion of a meal, three-fold above resting control values (approximately 300 micromol ml(-1)). This increase in plasma ammonia was accompanied by an increase in net ammonia flux to the water (approximately twofold higher in fed fish versus unfed fish). The arterial blood also became alkaline with increases in pH and plasma [HCO3-] between 3 and 12 h following feeding, representing the first measurement of an alkaline tide in a marine teleost. There was no evidence of respiratory compensation for the measured metabolic alkalosis, as Pa CO2 remained unchanged throughout the post feeding period. However, in contrast to an earlier study on freshwater-acclimated trout, fed fish did not exhibit a compensating increase in net base excretion, but rather took in additional base from the external seawater, amounting to approximately 8490 micromol kg(-1) over 48 h. In vitro experiments suggest that at least a portion of the alkaline tide was eliminated through increased HCO3- secretion coupled to Cl- absorption in the intestinal tract. This did not occur in the intestine of freshwater-acclimated trout. The marked effects of the external salinity (seawater versus freshwater) on different post-feeding patterns of acid-base balance are discussed. PMID- 19561206 TI - The effect of short- and long-term fasting on digestive and metabolic flexibility in the Andean toad, Bufo spinulosus. AB - Hibernation in ectothermic animals was historically considered as a simple cold induced torpor state resulting from the inability to maintain a high body temperature at low ambient temperatures. During the last decades this vision changed and nowadays there is a myriad of studies showing that hibernation implies different adjustments at the genetic, molecular, biochemical and cellular levels. However, studies oriented to evaluate changes of whole organism structure and physiology still are scarce, which is particularly true for amphibians that hibernate on land. Accordingly, in the Andean toad (Bufo spinulosus), we investigated the effect of short-term fasting and hibernation on the hydrolytic activity of digestive enzymes, histology of the small intestine, gross morphology of digestive and other internal organs and standard metabolic rate. Based on the pattern of size variation, internal organs may be grouped into those that were affected by both season and feeding condition (small intestine, stomach and liver), those that were only affected by season (fat bodies), those that were only affected by feeding condition (kidneys) and, finally, those that did not change between the three groups (large intestine, heart and lungs). Hydrolytic activity of maltase, trehalase and aminopeptidase-N followed the same pattern of variation (feeding > fasting > hibernating toads), although the change for the latter enzyme was less noticeable than for the disaccharidases. Enzymatic adjustments were correlated with changes in small intestine histology: villus and enterocyte height increased from hibernating to fasting and more markedly from fasting to feeding toads. Metabolic rate decreased during hibernation to 7.8% (at 5 degrees C) and 13.6% (at 15 degrees C) of summer values, which is one of the highest metabolic depressions reported for any ectothermic vertebrate. Our results suggest that amphibian persistence in highly seasonal environments is related to a large capacity of phenotypic flexibility at different organisational levels; an ability that may be related to the extensive ranges of temporal existence and geographic distribution of these vertebrates. PMID- 19561207 TI - Rapid clearance of circulating protein by early chicken embryo blood cells. AB - It has been speculated that free amino acids digested from proteins in bird eggs are transported to the circulation for the nourishment of the embryo. In the present study, we found that early chicken embryo protein in the serum might be utilized efficiently as a nutrient. Proteins injected into the blood of embryonic day 3 (E3) embryos were partially degraded and rapidly cleared. The rapid clearance of the injected proteins might be the result of efficient pinocytosis by blood cells, which then efficiently digested the intracellular proteins. An evaluation of the fluorescence intensity of injected fluorescein isothiocyanate labeled bovine serum albumin (FITC-BSA) indicated that about half was taken up by the blood cells 80 min after injection. About 4 h after injection, most of the FITC-BSA was digested and the products were released into the serum, which implies that circulating blood cells may serve as a digestive system in early chick embryos. However, the endocytic activity of blood cells decreased after E5, and BSA may reside in the circulation with a longer half-life after E5. These results imply that blood cells would serve as a digestive system only in early embryos. In summary, the mechanism revealed here gives the early embryo the ability to make use of protein as a nutrient without prior digestion outside the embryo. PMID- 19561208 TI - African lungfish, Protopterus annectens, possess an arginine vasotocin receptor homologous to the tetrapod V2-type receptor. AB - In tetrapods, arginine vasopressin and its counterpart, arginine vasotocin (AVT), are involved in renal water conservation through vascular V1a-type and tubular V2 type receptors, and only the former has thus far been cloned in fish. We successfully cloned the V1a-type and V2-type AVT receptor from the kidney of the African lungfish, Protopterus annectens, and the deduced amino acid sequences exhibited high homology with amphibian V1a- and V2-type receptors, respectively. Functional analysis showed that AVT addition to CHO cells transfected with lungfish V1a-type receptor increased [Ca2+]i in a concentration-dependent manner, whereas CHO cells transfected with lungfish V2-type receptor responded with cAMP accumulation after AVT stimulation. Lungfish V2-type receptor mRNA was strongly expressed in the heart and kidney, while V1a-type receptor mRNA was ubiquitously expressed in all the tissues examined. In the kidney, immunohistochemistry using a specific antibody to lungfish V2-type receptor showed localization in the basolateral area of the cells in the late part of the distal tubules. Artificial estivation (EST) for 90 days significantly increased plasma osmolality and sodium and urea concentrations. There was no significant difference in the V2-type receptor mRNA and protein expression levels in the kidney between the freshwater and EST lungfish, while the AVT precursor mRNA level in the hypothalamus was remarkably higher in the EST lungfish. Our results indicate that African lungfish possess a functional V2-type receptor similar to that in tetrapods, suggesting that elevated plasma AVT during estivation exerts a renal tubular antidiuretic effect through the V2-type receptor expressed in the distal segments of lungfish kidney. PMID- 19561209 TI - Cleaner gobies evolve advertising stripes of higher contrast. AB - Elacatinus gobies of the Caribbean have undergone rapid speciation along ecological axes, and particular species from this genus act as 'cleaners' that remove ectoparasites from larger coral reef fish, termed 'clients'. Evolutionary shifts in habitat use, behavior and lateral body stripe colors differentiate cleaners from ancestral sponge-dwelling lineages. High-contrast stripe colors associated with cleaning behavior on coral reefs may have evolved as a signal of cleaning status. We asked whether cleaner gobies with blue stripes are more conspicuous than ancestral yellow- and green-stripe phenotypes to a diverse set of potential client visual systems in the tropical reef environment where cleaning stations are commonly observed. Using spectrophotometric measurements of cleaners with blue and yellow stripes and their F1 hybrid, we tested the contrast of each color stripe to both potential dichromatic and trichromatic reef fish visual systems, against typical coral and sponge microhabitat background colors. Blue stripes provide the highest average chromatic contrast across a range of possible microhabitat colors to the majority of fish visual systems tested. The contrast provided by yellow and hybrid green stripes are comparable across habitats to dichromatic visual systems. The green stripe is less contrasting than both blue and yellow to many potential trichromatic visual systems. We suggest that the evolution of blue stripes in Elacatinus gobies could be a result of natural selection for signals of high color contrast, driven by the sensory biases and visual systems of diverse reef fish clients. PMID- 19561210 TI - Function of the sexually dimorphic ear of the American bullfrog, Rana catesbeiana: brief review and new insight. AB - The dimorphic ear of the bullfrog, Rana catesbeiana, has long been enigmatic. The male's tympanic membrane (TM) area approximates twice the area of the female's; however, similar size differences in the area of the columellar footplate were not observed between the sexes. Hence, the male's hearing is expected to be more sensitive than the female's but this is not the case. Asking what offsets the advantage of the large TM, we applied a series of experiments to the auditory system. Male and female audiograms based on stimulation with airborne sound and on both multi-unit responses from the brain and alternating cochlear potentials ('microphonics') showed equal sensitivity and a small difference in frequency response; at low frequencies the male was more sensitive than the female. Amputating the columella and stimulating the stump with mechanical vibration showed that for an equal microphonic response, the male's footplate vibrated with lower amplitude than the female's footplate. Mechanically stimulating the TM of the intact ear replicated this result, excluding the involvement of the mechanical lever. The TM of the male weighs five times the TM of the female, and artificial loading of the TM of either sex greatly reduced the ear's sensitivity. Hence, the male's excessive area ratio (TM to columellar footplate) is offset by the heavier cartilage cushion on the male's TM, damping the TM's response to sound. This is corroborated by experimentally artificially loading the TM. The product of area ratio and footplate vibration amplitude would result in similar stimulation of the inner ear in the two sexes. PMID- 19561211 TI - A study of synchronisation between the flagella of bull spermatozoa, with related observations. AB - Flagellar synchronisation has been observed between bull spermatozoa as they swam in a viscous medium, confined to a glass surface. This process is of interest in understanding the regulation of flagellar oscillation in general. Exact and persisting synchrony between bull spermatozoa occurred only when the spermatozoan heads were tightly coupled mechanically. For these cells, viscous coupling between the flagella was not by itself sufficient to establish synchronisation. Immediately on synchronisation, with the spermatozoan heads superposed, the paired spermatozoa showed rises in conjoint beat frequency, wave velocity and swimming velocity, i.e. in nearly all cases, the new conjoint values were greater than those shown by either of the two singleton spermatozoa. In our interpretation of these results, we put forward hydrodynamic arguments for seeing the primary change as a rise in wave velocity, via a decreased viscous resistance to bend propagation. Mechanistically, the rise in beat frequency is mysterious unless, as we suggest, it is consequential to the rise in wave velocity, and mediated by an as-yet-unknown mechanical feedback process. The rise in swimming velocity is not surprising given the rise in wave velocity but there is evidence for an additional influence due to a subtle re-orientation of the conjoint spermatozoan heads, such that they experienced less frictional drag. PMID- 19561212 TI - Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis. AB - The marine mussel Mytilus galloprovincialis is tethered to rocks in the intertidal zone by a holdfast known as the byssus. Functioning as a shock absorber, the byssus is composed of threads, the primary molecular components of which are collagen-containing proteins (preCOLs) that largely dictate the higher order self-assembly and mechanical properties of byssal threads. The threads contain additional matrix components that separate and perhaps lubricate the collagenous microfibrils during deformation in tension. In this study, the thread matrix proteins (TMPs), a glycine-, tyrosine- and asparagine-rich protein family, were shown to possess unique repeated sequence motifs, significant transcriptional heterogeneity and were distributed throughout the byssal thread. Deamidation was shown to occur at a significant rate in a recombinant TMP and in the byssal thread as a function of time. Furthermore, charge heterogeneity presumably due to deamidation was observed in TMPs extracted from threads. The TMPs were localized to the preCOL-containing secretory granules in the collagen gland of the foot and are assumed to provide a viscoelastic matrix around the collagenous fibers in byssal threads. PMID- 19561213 TI - 'Different strokes for different folks': geographically isolated strains of Lymnaea stagnalis only respond to sympatric predators and have different memory forming capabilities. AB - Gaining insight into how natural trait variation is manifest in populations shaped by differential environmental factors is crucial to understanding the evolution, ecology and sensory biology of natural populations. We have demonstrated that lab-reared Lymnaea detect and respond to the scent of a crayfish predator with specific, appropriate anti-predator behavioral responses, including enhanced long-term memory (LTM) formation, and that such predator detection significantly alters the electrophysiological activity of RPeD1, a neuron that is a necessary site for LTM formation. Here we ask: (1) do distinct populations of wild Lymnaea stagnalis respond only to sympatric predators and if so, can these traits be quantified at both the behavioral and neurophysiological levels, and (2) does the presence of a non-sympatric predator elicit anti predator behaviors including augmentation of LTM? We tested three different populations of wild (i.e. not lab-reared) snails freshly collected from their natural habitat: (1) polders near Utrecht in The Netherlands, (2) six seasonally isolated ponds in the Belly River drainage in southern Alberta, Canada and (3) a 20-year-old human-made dugout pond in southern Alberta. We found strain-specific variations in the ability to form LTM and that only a sympatric predator evoked anti-predatory behaviors, including enhanced LTM formation and changes in RPeD1 activity. PMID- 19561214 TI - Surviving the drought: burrowing frogs save energy by increasing mitochondrial coupling. AB - During dormancy energy conservation is a key priority and as such dormant animals undergo a major metabolic depression to conserve their limited endogenous fuel supplies. Mitochondrial coupling efficiency, the efficiency with which mitochondria convert oxygen into ATP, significantly affects aerobic metabolism and thus to maximise energy savings during dormancy it has been hypothesised that coupling efficiency should increase. However, previous studies have shown coupling efficiency to be maintained or even to decrease. In this study we measured state 3 and state 4 mitochondrial respiration in the muscle of the burrowing frog, Cyclorana alboguttata and calculated the respiratory control ratio as a measure of coupling efficiency. After 7 months in aestivation, C. alboguttata significantly reduced oxygen consumption of isolated mitochondria by 83% and, unlike other dormant animals, the frogs appeared to decrease rates of proton leak to a greater extent than ATP synthesis, consistent with an increase in mitochondrial coupling efficiency. The significant energy savings observed at the mitochondrial level were reflected at higher levels of biological organisation, with tissue oxygen consumption depressed by as much as 81% and whole animal metabolic rate by 82%. Cyclorana alboguttata can survive in a dormant state for several years and we propose the hypothesis that energy efficiency is increased during aestivation. PMID- 19561215 TI - Targets for TNFalpha-induced lipolysis in gilthead sea bream (Sparus aurata L.) adipocytes isolated from lean and fat juvenile fish. AB - The present study aimed to analyze adiposity heterogeneity and the role of liver X receptor (LXRalpha) and peroxisome proliferator-activated receptors (PPARs) as targets of tumour necrosis factor-alpha (TNFalpha) in gilthead sea bream (Sparus aurata L.). The screening of 20 fish at the beginning of the warm season identified two major groups with fat and lean phenotypes. Fat fish showed increased liver and mesenteric fat depots. This increased adiposity was concurrent in the adipose tissue to enhanced expression of lipoprotein lipase (LPL) whereas mRNA levels of the hormone-sensitive lipase (HSL) remained almost unchanged. The resulting LPL/HSL ratio was thereby highest in fat fish, which suggests that this group of fish has not reached its peak fat storage capacity. This is not surprising given the increased expression of PPARgamma in the absence of a counter-regulatory raise of TNFalpha. However, this lipolytic cytokine exerted dual effects in primary adipocyte cultures that differ within and between lean and fat fish. One set of fat fish did not respond to TNFalpha treatment whereas a second set exhibited a lipolytic response (increased glycerol release) that was apparently mediated by the downregulated expression of PPARbeta. In lean fish, TNFalpha exerted a strong and non-transcriptionally mediated lipolytic action. Alternatively, TNFalpha would inhibit lipid deposition via the downregulated expression of adipogenic nuclear factors (PPARgamma and LXRalpha). TNFalpha targets are therefore different in fish with lean and fat phenotypes, which is indicative of the complex network involved in the regulation of fish lipid metabolism. PMID- 19561216 TI - A supracellular system of actin-lined canals controls biogenesis and release of virulence factors in parasitoid venom glands. AB - Parasitoid wasps produce virulence factors that bear significant resemblance to viruses and have the ability to block host defense responses. The function of these virulence factors, produced predominantly in wasp venom glands, and the ways in which they interfere with host development and physiology remain mysterious. Here, we report the discovery of a specialized system of canals in venom glands of five parasitoid wasps that differ in their infection strategies. This supracellular canal system is made up of individual secretory units, one per secretory cell. Individual units merge into the canal lumen. The membrane surface of the proximal end of each canal within the secretory cell assumes brush border morphology, lined with bundles of F-actin. Systemic administration of cytochalasin D compromises the integrity of the secretory unit. We show a dynamic and continuous association of p40, a protein of virus-like particles from a Drosophila parasitoid, L. heterotoma, with the canal and venom gland lumen. Similar structures in three Leptopilina species and Ganaspis xanthopoda, parasitoids of Drosophila spp., and Campoletis sonorenesis, a parasitoid of Heliothis virescens, suggest that this novel supracellular canal system is likely to be a common trait of parasitoid venom glands that is essential for efficient biogenesis and delivery of virulence factors. PMID- 19561217 TI - Visualization of the spatial and spectral signals of orb-weaving spiders, Nephila pilipes, through the eyes of a honeybee. AB - It is well known that the honeybee has good color vision. However, the spectral range in which the bee can see is different from that of the human eye. To study how bees view their world of colors, one has to see through the eyes of the bee, not the eyes of a human. A conventional way to examine the color signals that animals can detect is to measure the surface reflectance spectra and compute the quantum catches of each photoreceptor type based on its known spectral sensitivity. Color signal and color contrast are then determined from the loci of these quantum catches in the color space. While the point-by-point measurements of the reflectance spectra using a standard spectrometer have yielded a significant amount of data for analyzing color signals, the lack of spatial information and low sampling efficiency constrain their applications. Using a special filter coating technique, a set of filters with transmission spectra that were closely matched to the bee's sensitivity spectra of three photoreceptor types (UV, blue, and green) was custom made. By placing these filters in front of a UV/VIS-sensitive CCD camera and acquiring images sequentially, we could collect images of a bee's receptor with only three shots. This allowed a direct visualization of how bees view their world in a pseudo-color RGB display. With this imaging system, spatial and spectral signals of the orb-weaving spider, Nephila pilipes, were recorded, and color contrast images corresponding to the bee's spatial resolution were constructed and analyzed. The result not only confirmed that the color markings of N. pilipes are of high chromatic contrast to the eyes of a bee, but it also indicated that the spatial arrangement of these markings resemble flower patterns which may attract bees to visit them. Thus, it is likely that the orb-weaving spider (N. pilipes) deploys a similar strategy to that of the Australian crab spider (Thomisus spectabilis) to exploit the bee's pre-existing preference for flowers with color patterning. PMID- 19561218 TI - Mudskipper pectoral fin kinematics in aquatic and terrestrial environments. AB - Mudskippers use pectoral fins for their primary mode of locomotion on land and pectoral fins in conjunction with the axial musculature and caudal fin to move in water. We hypothesized that distinct pectoral fin movements enable effective locomotion in each environment. Additionally, we made three functional predictions about fin movements during locomotion on land versus water: the pectoral fin is depressed more on land than in water; the pectoral fin will have greater changes in fin area between propulsive and recovery phases in water versus land; anterior and posterior excursions will be greater on land than in water. Locomotion was recorded in each environment using a high-speed digital imaging system and kinematic variables were calculated from digitized landmark points. Variables were analyzed using principal components analysis and matched pairs t-tests. Mudskippers produce distinct kinematic patterns across environments (P < 0.003), although only some of our predictions were supported. The magnitude of fin depression is the same across habitats. However, depression occurs during the propulsive phase on land (by -0.60 cm), whereas during the propulsive phase in water the fin is elevated (by +0.13 cm). We were unable to support the hypothesis that fin orientation differs between environments. Lastly, anterior extension of the fin is greater on land (1.8 cm, versus 1.3 cm in water), creating a larger stride length in this environment. We posit that the mudskipper pectoral fin may facilitate stability in water and thrust production on land, and suggest that the robust fin morphology of the goby lineage may predispose species within this group to terrestrial locomotion. PMID- 19561219 TI - Limiting factors to encapsulation: the combined effects of dissolved protein and oxygen availability on embryonic growth and survival of species with contrasting feeding strategies. AB - Encapsulation is a common strategy among marine invertebrate species. It has been shown that oxygen and food availability independently constrain embryo development during intracapsular development. However, it is unclear how embryos of species with different feeding strategies perceive these two constraints when operating jointly. In the present study, we examined the relative importance of dissolved albumen, as a food source, oxygen condition and their interaction on embryonic growth and the survival of two calyptraeid species, Crepidula coquimbensis and Crepidula fornicata, exhibiting different embryo feeding behaviours (i.e. presence vs absence of intracapsular cannibalism). Two oxygen condition treatments (normoxia and hypoxia) and three albumen concentrations (0, 1 and 2 mg l(-1)) were studied. In addition, albumen intake by embryos was observed using fluorescence microscopy. Our study shows that embryos of both species incorporated dissolved albumen but used a different set of embryonic organs. We observed that embryo growth rates in C. coquimbensis were negatively affected only by hypoxic conditions. Conversely, a combination of low albumen concentration and oxygen availability slowed embryo growth in C. fornicata. These findings suggest that oxygen availability is a limiting factor for the normal embryo development of encapsulated gastropod species, regardless of feeding behaviour or developmental mode. By contrast, the effect of dissolved albumen as an alternative food source on embryo performance may depend on the feeding strategy of the embryos. PMID- 19561220 TI - The behavioural, digestive and metabolic characteristics of fishes with different foraging strategies. AB - To test the hypothesis that digestion has a more notable physiological effect on ambush foragers than on active foragers, we investigated the behavioural, digestive and metabolic characteristics, as well as the postprandial locomotory capacity, of four species of juvenile fish distributed along the Yangtze River, China, with distinct foraging strategies. The ambush foraging southern catfish (Silurus meridionlis) had the fewest movements per minute (MPM), lowest per cent time spent moving (PTM), slowest critical swimming speed (Ucrit), lowest maintenance metabolism (VO2rest) and lowest maximum locomotory metabolism (VO2max)). However, the southern catfish had the highest feeding level and maximum feeding metabolism (VO2peak) and the greatest decrease in Ucrit after consumption of a large meal. Thus, this fish is highly adapted to its ambush behavioural strategy and sedentary life style. In the herbivorous grass carp (Ctenopharyngodon idellus), a low digestive capacity led to little change in postprandial locomotory performance, which benefits its frequent grazing behaviour. In this species, the greater amount of energy spent on routine activity and avoiding predators versus Ucrit might be related to its herbivorous life style and high predation risk. The active foraging crucian carp (Carassius auratus) adopts a unique high energy cost strategy that allows for high capacity in both routine activity and digestion, and the great flexibility of its cardio respiratory capacity (increased VO2max after feeding) guarantees a small decrease in Ucrit even after maximum feeding. Finally, the sluggish foraging darkbarbel catfish (Pelteobagrus vachelli) has low digestive and locomotory capacity, but its energy-efficient venomous defence strategy may be related to its abundance. These results show that the digestive, behavioural and metabolic strategies differ among these fish species. The locomotory capacity in the sedentary fishes decreased profoundly after feeding, whereas it decreased little or not at all in the active fishes. The maintenance of high locomotory capacity after eating in the active fishes is probably related to a large metabolic capacity, a lower digestive capacity or an improvement in cardio-respiratory capacity after feeding. PMID- 19561227 TI - Cardiac pathology of systemic lupus erythematosus. AB - Systemic lupus erythematosus is a common chronic autoimmune disorder causing injury to many organ systems. Cardiac complications of lupus affect most parts of the heart. These include pericarditis, myocarditis, endocarditis and coronary artery disease. While many histopathological findings in lupus-related cardiac diseases are non-specific, there are a few important findings which pathologists should be aware of. This review provides pathological descriptions of these entities. PMID- 19561221 TI - A patient with a leg rash, pedal edema, renal failure, and thrombocytopenia. PMID- 19561228 TI - Clinical audit in the laboratory. AB - Audits are part of the continuous quality improvement process and one of the key elements of clinical governance. Laboratory-based clinical audits are concerned primarily with the everyday aspects of laboratory services and are a means of providing feedback to the users of the laboratory and its staff. They involve measuring the performance of laboratory services against established standards. These standards have ideally been established using the principles of evidence based medicine. If necessary, changes are implemented and then a re-audit is performed after a certain time period to ensure that the changes have been implemented and maintained. Areas of audit in the laboratory include the preanalytical, analytical and postanalytical phases. This review article examines the basis of clinical audits in the laboratory and then proceeds to describe in detail how a laboratory-based clinical audit should be performed and monitored, with special reference to the chemical pathology laboratory. PMID- 19561229 TI - Genomic profile of a secretory breast cancer with an ETV6-NTRK3 duplication. AB - BACKGROUND: Secretory breast cancer (SBC) is a rare entity characterised by indolent clinical behaviour, distinctive histological features and the presence of a recurrent chromosomal translocation t(12;15)(p13;q25), leading to the formation of the ETV6-NTRK3 fusion gene. AIM: To describe the molecular genetic features of a case of SBC which harbours a duplication of the t(12;15) translocation. METHODS: Tiling path array comparative genomic hybridisation (aCGH) analysis and fluorescence in situ hybridisation (FISH) using in-house generated probes for ETV6, NTRK3 and the fusion genes, centromeric probes for chromosomes 12 and 15, and a commercially available split-apart ETV6/NTRK3 probe. RESULTS: FISH revealed the presence of a duplication of the translocation t(12;15), which resulted from the gain of one copy of the derivative chromosome der(15)t(12;15), retention of one normal copy of both ETV6 and NTRK3 genes and deletion of the derivative chromosome der(12)t(12;15). Consistent with FISH findings, aCGH revealed copy number gains of ETV6 and NTRK3 and deletions encompassing the regions centromeric to ETV6 and telomeric to NTRK3. Additional regions of copy number changes included gains of 10q21, 10q26.3, 12p13.3-p13.31 15q11-q25.3 and 16pq and losses of 6q24.1-q27, 12p13.2-q12 and 15q25.3-q26.3. CONCLUSIONS: To the best of our knowledge, this is the first time a carcinoma has been shown to harbour a duplication of the ETV6-NTRK3 translocation. The presence of an additional copy of the derivative chromosome der(15)t(12;15) coupled with deletion of the other derivative der(12)t(12;15) in the modal population of cancer cells suggests that this was either an early phenomenon or conferred additional growth advantage on neoplastic cells. PMID- 19561230 TI - V600E BRAF mutations are alternative early molecular events in a subset of KIT/PDGFRA wild-type gastrointestinal stromal tumours. AB - BACKGROUND: A small subset (10-15%) of gastrointestinal stromal tumours (GISTs) lack mutations in KIT and PDGFRA (wild-type GIST). Recently, a novel BRAF exon 15 mutation (V600E) was detected in imatinib-naive wild-type high-risk intestinal GISTs (4%). However, the frequency and distribution of BRAF mutations within the spectrum of GISTs, and whether they might represent secondary events acquired during tumour progression, remain unknown. METHODS: 69 GISTs (39 KIT mutants, 2 PDGFRA mutants and 28 wild-type) were analysed for mutations in BRAF exon 15 and KRAS exon 2. To assess the stage at which these mutations might occur in GIST, a considerable number of incidental gastric (n = 23) and intestinal (n = 2) tumours were included. RESULTS: BRAF mutations (V600E) were detected in 2 of 28 wild-type GISTs (7%), but in none of the 41 KIT/PDGFRA mutants. No KRAS mutation was detected. The two BRAF-mutated GISTs measured 4 mm in diameter and originated in the gastric body and the jejunum in two men (mean age, 76 years). Both tumours were mitotically inactive KIT-positive spindle-cell GISTs that were indistinguishable histologically from their more common KIT-mutated counterparts. CONCLUSION: BRAF mutations represent an alternative molecular pathway in the early tumorigenesis of a subset of KIT/PDGFRA wild-type GISTs and are per se not associated with a high risk of malignancy. Mutations in KIT, PDGFRA and BRAF were mutually exclusive in this study. Results from this and a previous study indicate that BRAF-mutated GISTs show a predilection for the small bowel (four of five tumours), but this needs further evaluation in larger studies. PMID- 19561231 TI - Triple negative breast cancer: a study from the point of view of basal CK5/6 and HER-1. AB - AIM: Basal-like breast tumours, as defined by microarrays, carry a poor prognosis and therapeutic options are limited to date. Often, these tumours are defined as oestrogen receptor (ER) negative/progesterone receptor (PR) negative/human epidermal growth factor receptor 2 (HER-2) negative (triple negative) by immunohistochemistry (IHC), but a more complete definition should include expression of basal cytokeratins (CK5/6, CK14 or CK17) and/or human epidermal growth factor receptor 1 (HER-1). The aim of this study was to investigate to what extent CK5/6 and HER-1 characterise the group of triple negative breast cancers. METHODS: Expression of CK5/6 and HER-1 was studied by IHC in 25 triple negative breast carcinomas and 32 grade-matched, non-triple-negative controls. All 57 cases were further subjected to fluorescence in situ hybridisation to investigate HER-1 gene copy number. RESULTS: CK5/6 and HER-1 expression was most frequent in triple negative tumours: 22 out of 25 cases (88.0%) expressed at least one of these markers (60.0% CK5/6 positive and 52.0% HER-1 positive). In the control group, CK5/6 and HER-1 expression was found in ER-negative but not in ER-positive tumours (ER negative/PR negative/HER-2 positive tumours: 20.0% CK5/6 positive and 46.7% HER-1 positive). HER-1 gene amplification was found in five cases only: four triple negative (16.0%) and one ER-negative control (ER negative/PR negative/HER-2 positive, 6.7%). Of interest, all five HER-1 amplified cases showed a remarkably homogeneous HER-1 expression pattern. CONCLUSION: Expression of CK5/6 and HER-1 is frequent in ER-negative breast cancers, in triple negative and in non-triple negative tumours. In a minority of cases, HER-1 overexpression may be caused by HER-1 gene amplification. Further studies are needed to investigate whether such cases might benefit from anti-HER-1 therapy. PMID- 19561232 TI - Cytological features of melanoma in exfoliative fluid specimens. AB - AIMS: To describe the cytological appearances of melanoma in fluid specimens and potential diagnostic pitfalls in interpreting such specimens, which have been infrequently reported in the literature. METHODS: Cases of melanoma diagnosed at a single institution between 1993 and 2008 in cytology specimens of fluids (pleural, ascitic, cerebrospinal and other fluids), but excluding fine needle biopsy specimens, were identified and reviewed. RESULTS: 32 fluid specimens containing metastatic melanoma (from 26 patients) were identified. Most of the specimens were moderately cellular and showed moderate to marked nuclear pleomorphism. Mitotic figures and intranuclear cytoplasmic invaginations were identified in 11 (34.4%) and seven (21.9%) cases, respectively. Melanin pigment was seen in eight (25.0%) cases. Variable numbers of histiocytes were present, and mesothelial cells were present in body cavity fluid specimens. CONCLUSIONS: In fluid specimens, reactive mesothelial cells and histiocytes may mimic epithelioid melanoma cells. Awareness of the morphological features and diagnostic pitfalls of melanoma in fluids is necessary to avoid the potentially serious consequences of misdiagnosis. PMID- 19561233 TI - Vulvar plasmablastic lymphoma in a HIV-positive child: a novel extraoral localisation. AB - Plasmablastic lymphoma (PBL) has been characterised by the World Health Organization as a new entity. This report describes an unusual case of PBL in a 3 year-old HIV-infected patient showing a cutaneous vulvar lesion with 9 months of evolution and prolapsed vulvovaginal mucosa. Histopathological examination of a biopsy sample showed diffuse submucosal infiltration by large cells with a cohesive growth pattern, and round and vesicular nuclei with fine chromatin centrally or eccentrically placed with one or more prominent nucleoli. Immunohistochemical staining in neoplastic cells was positive for multiple melanoma oncogene (MUM1), CD138, CD45 and epithelial membrane antigen (EMA). The diagnosis was PBL, stage III. Epstein-Barr virus (EBV) expression was positive by EBV encoded RNAs in situ hybridisation. This is believed to be the third case of paediatric HIV-associated PBL reported in the literature, and the first with vulvar localisation, which is a new anatomical location for this entity. PMID- 19561234 TI - Spontaneous regression of natural killer cell lymphoma. AB - Spontaneous tumour regression is extremely rare in aggressive lymphoma. A case of natural killer (NK) cell lymphoma with cutaneous manifestation showed an indolent clinical course, and the relapsed nodular lesion disappeared spontaneously without any treatment. Although only small number of T cells were present in the primary skin lesion, there was massive CD8-positive cytotoxic T cell infiltration in the relapsed lesion. This is believed to be the first report of an abscopal effect on NK cell lymphoma. Infiltration of cytotoxic T cells strongly suggests immunological attack against the lymphoma cells. PMID- 19561235 TI - An EMA negative, desmin positive malignant mesothelioma: limitations of immunohistochemistry? AB - Histopathologists in the current environment of medical negligence and litigation are more likely to use immunohistochemical investigations in their day-to-day practice to support their diagnosis and avoid future litigation. The caveat is that relying on immunohistochemistry is a double-edged sword and pathologists should be familiar with its limitations. We present a case of primary malignant peritoneal mesothelioma with an unusual immunohistochemical profile-desmin positive, EMA negative-and wish to highlight the importance of cautiously interpreting immunohistochemistry profiles when they do not fit the clinical history and histological appearance. PMID- 19561236 TI - Malignant myoepithelioma arising in adenomyoepithelioma of the breast and coincident multiple gastrointestinal stromal tumours in a patient with neurofibromatosis type 1. AB - A 41-year-old female patient with neurofibromatosis type 1 (NF-1) presented with a breast lump and anaemia related to gastrointestinal bleeding. She was found to have malignant myoepithelioma of the breast and simultaneously multiple gastrointestinal stromal tumours (GISTs) of the small bowel. Molecular studies showed a silent germline mutation in exon 9 of the KIT gene of both tumours. The common gene mutations characteristic of sporadic GISTs were not identified in these tumours, consistent with the literature, suggesting that gene mutations in GISTs are either absent or late events in patients with NF-1. PMID- 19561237 TI - An unusual case of Epstein-Barr virus driven lymphoproliferative disorder of the conjunctiva which mimicked a high grade lymphoma: a sheep in wolf's clothing. AB - Lymphoid proliferations represent 25-33% of acquired sub-epithelial conjunctival lesions which are excised or biopsied in patients over 15 years of age. These lesions are reported in association with Epstein-Barr virus (EBV). One such case of EBV associated spontaneously regressed monoclonal B cell infiltrate in conjunctiva that mimicked a large B cell lymphoma is reported. PMID- 19561238 TI - Composite phaeochromocytoma with malignant peripheral nerve sheath tumour and rhabdomyosarcomatous differentiation in a patient without von Recklinghausen disease. AB - The coexistence of adrenal phaeochromocytoma with non-chromaffin tumours is a rare fascinating occurrence. This category of tumours is subdivided into "composite" and "mixed". The coexistence of adrenal phaeochromocytoma with a malignant Triton tumour does not appear to have been described in the available literature so far. A unique case of composite phaeochromocytoma in a 26-year-old male patient, where the non-chromaffin component was a malignant Triton tumour composed of peripheral nerve sheath tumour and skeletal muscle differentiation, is reported. This admixture was confirmed with immunohistochemical pattern of expression. This is the first case of such a phenomenon in a composite phaeochromocytoma. The present case further widens the histomorphological range of composite phaeochromocytoma of the adrenal gland, which the histopathologist should be aware of. Since the prognosis of composite phaeochromocytoma with malignant nerve sheath tumour would be determined by the nerve sheath component, recognition of this tumour is imperative. PMID- 19561239 TI - Soluble CD30 as a prognostic factor for outcome following renal transplantation. AB - AIMS: To determine whether measurement of soluble CD30 (sCD30) levels predicts for early rejection in a cohort of first deceased kidney transplant recipients. METHODS: Pre-transplant serum samples were analysed for sCD30 levels using a commercial ELISA kit (Biotest). A 100 U/ml cut-off for "high sCD30" was applied. Clinical outcome parameters were biopsy-proven rejection episodes, creatinine levels and glomerular filtration rate. RESULTS: In the cohort of patients who experienced at least one episode of rejection in the first 6 months post transplant, levels of pre-transplant sCD30 were significantly higher than in those who did not experience rejection. Despite this association, the occurrence of a high sCD30 level did not predict for rejection on an individual basis. CONCLUSIONS: The prognostic value of pre-transplant sCD30 testing is diminished by the large number of patients with high sCD30 levels who do not develop rejection. Although this limits the utility of the test in informing clinical management of individual patients, a high pre-transplant sCD30 level should still be considered a risk factor for poorer outcome. PMID- 19561240 TI - Quality of teaching in chemical pathology: ability of interns to order and interpret laboratory tests. AB - BACKGROUND AND OBJECTIVE: There has been a steady decline in the overt teaching of many basic and pathology sciences in the medical curriculum worldwide. As interns are the doctors most likely to request and act on tests, an assessment of their confidence in dealing with laboratory investigations was undertaken. METHODS: Interns at two hospitals in Cape Town, South Africa, were asked to complete a structured questionnaire designed to assess their confidence in ordering and interpreting a number of tests. The questionnaire also probed their desire for further teaching and the preferred delivery vehicle for such teaching. RESULTS AND CONCLUSIONS: 61 out of 117 questionnaires were returned. Interns were confident in the use of common tests, but 23% were not confident in interpreting a test that they were confident in ordering. All respondents felt they would benefit from teaching in at least one area and lectures were the preferred method, although the majority felt it very likely that they would complete an online tutorial if available. The results suggest that institutions need to devise strategies to fulfil the learning needs of new graduates in the area of chemical pathology and clinical biochemistry. PMID- 19561242 TI - Average serum triglyceride concentration in relation to day of the week in Burton on-Trent. PMID- 19561243 TI - Oesophageal rupture due to tuberculous pseudoaneurysm of the aorta. PMID- 19561244 TI - Can imaging gene expression in human mesenchymal stem cells be successful in large animals? PMID- 19561246 TI - Radiation, CT, and children: the simple answer is ... it's complicated. PMID- 19561247 TI - Tracheobronchomalacia in infants and children: multidetector CT evaluation. AB - Tracheobronchomalacia (TBM) is the most common congenital central airway anomaly, but it frequently goes unrecognized or is misdiagnosed as other respiratory conditions such as asthma. Recent advances in multidetector computed tomography (CT) have enhanced the ability to noninvasively diagnose TBM with the potential to reduce the morbidity and mortality associated with this condition. Precise indications are evolving but may include symptomatic pediatric patients with known risk factors for TBM and patients with otherwise unexplained impaired exercise tolerance; recurrent lower airways infection; and therapy-resistant, irreversible, and/or atypical asthma. With multidetector CT, radiologists can now perform objective and quantitative assessment of TBM with accuracy similar to that of bronchoscopy, the reference standard for diagnosing this condition. Multidetector CT enables a comprehensive evaluation of pediatric patients suspected of having TBM by facilitating accurate diagnosis, determining the extent and degree of disease, identifying predisposing conditions, and providing objective pre- and postoperative assessments. In this article, the authors present a step-by-step primer of multidetector CT imaging for evaluating infants and children with suspected TBM, including clinical indications, patient preparation, multidetector CT techniques and protocols, two- and three dimensional processing of multidetector CT data, and image interpretation. The major aim of this article is to facilitate the reader's ability to successfully employ multidetector CT imaging protocols for evaluation of TBM in infants and children in daily clinical practice. PMID- 19561248 TI - Net Assets: The social Web for radiology. Part I. Social bookmarking to social citation. PMID- 19561249 TI - Gerbode ventricular septal defect diagnosed at cardiac MR imaging: case report. AB - This report describes the ability of cardiac magnetic resonance (MR) imaging to depict an unusual cardiac defect. A type of ventricular septal defect called the Gerbode defect, which results in a communication between the left ventricle and the right atrium, is presented. To the authors' knowledge, this is the first time cardiac MR imaging has been utilized to characterize this defect. PMID- 19561250 TI - Pulmonary embolism detection with dual-energy CT: experimental study of dual source CT in rabbits. AB - PURPOSE: To evaluate feasibility and added value of dual-energy computed tomography (CT) in diagnosis of pulmonary embolism (PE). MATERIALS AND METHODS: This institutional animal experimental committee-approved study was performed in accordance with animal care guidelines. Eight New Zealand rabbits underwent standard unenhanced and contrast material-enhanced dual-source CT. Gelatin sponge particles were injected into the pulmonary artery, and rabbits underwent contrast enhanced dual-source CT pulmonary angiography, from which blood-flow (BF) and fusion images were created. Immediately after dual-source CT, rabbits were sacrificed, their lungs were removed and fixed in 10% formalin, and detailed pathologic determination of location and number of lung lobes with PE was performed. Two rabbits were excluded: One died during the procedure. In the other, the catheter tip was retained in the left inferior pulmonary artery. This caused marked postembolization CT image artifacts in adjacent regions. Six rabbits were included in final analysis. Two radiologists without knowledge of pathologic results evaluated five pulmonary lobes in each rabbit and recorded whether PE was present. Pathologic results served as the reference standard. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the techniques were calculated. Weighted kappa values were calculated to evaluate agreement between modalities. RESULTS: Pathologic analysis revealed PE in 18 of 30 pulmonary lobes. Conventional CT angiography was used to correctly identify PE in 12 lobes and absence of emboli in 18 lobes, which corresponded to sensitivity, specificity, PPV, and NPV of 67%, 100%, 100%, and 67%, respectively. A kappa value of 0.65 indicated good correlation with pathologic findings. On BF images, segments with an embolic region showed low perfusion compared with segments with a normal pulmonary region. BF images and fused images correctly showed PE in 16 of 18 pulmonary lobes and absence of emboli in 11 of 12 lobes, which corresponded to sensitivity, specificity, PPV, and NPV of 89%, 92%, 94%, and 85%, respectively, in detection of PE. A kappa value of 0.80 indicated good correlation with pathologic findings. CONCLUSION: Dual-source CT can depict normal and abnormal blood perfusion distribution in a rabbit's lung. Abnormal pulmonary blood distribution, as shown at dual-source CT, improves detection of acute PE in rabbits. PMID- 19561251 TI - Ectopic pancreas: CT findings with emphasis on differentiation from small gastrointestinal stromal tumor and leiomyoma. AB - PURPOSE: To describe the computed tomographic (CT) findings of ectopic pancreas and to identify the features that differentiate it from other similarly manifesting gastric submucosal tumors such as gastrointestinal stromal tumor (GIST) and leiomyoma, which are the most common gastrointestinal submucosal tumors. MATERIALS AND METHODS: This retrospective study was approved by the institutional review board and did not require informed consent. CT images of pathologically proved ectopic pancreases (n = 14), GISTs (n = 33), and leiomyomas (n = 7) in the stomach and duodenum were retrospectively reviewed. Analysis of the CT findings included evaluation of the location, contour, growth pattern, border, enhancement pattern, and enhancement grade of the tumor, as well as the presence of surface dimpling, prominent enhancement of overlying mucosa, and low intralesional attenuation. The attenuation of each lesion, the long diameter (LD), the short diameter (SD), and the LD/SD ratio were measured. Among these findings, statistically significant variables were determined by using the chi(2) test (to compare the categoric variables), the Student t test (for quantitative analysis), and the receiver operating characteristic curve (to determine the optimal cutoff of the LD/SD ratio). RESULTS: The typical location (prepyloric antrum and duodenum), endoluminal growth pattern, ill-defined border, prominent enhancement of overlying mucosa, and an LD/SD ratio of greater than 1.4 were found to be significant for differentiating ectopic pancreas from other tumors (P < .05 for each finding). When at least two of these five criteria were used in combination, the sensitivity and specificity for diagnosing ectopic pancreas were 100% (14 of 14) and 82.5% (33 of 40), respectively. When four of these criteria were used, a sensitivity of 42.9% and a specificity of 100% were achieved. CONCLUSION: By using specific CT criteria, ectopic pancreas can be differentiated from small GIST or leiomyoma with a high degree of accuracy. PMID- 19561252 TI - Prostate MR imaging: tissue characterization with pharmacokinetic volume and blood flow parameters and correlation with histologic parameters. AB - PURPOSE: To prospectively determine whether pharmacokinetic magnetic resonance (MR) imaging parameters correlate with histologic mean vessel density (MVD), mean vessel area (MVA), and mean interstitial area (MIA) and whether these parameters enable differentiation of prostate cancer, chronic prostatitis, and normal prostate tissue. MATERIALS AND METHODS: This study was approved by the institutional review board, and informed consent was obtained from all patients. Thirty-five patients with biopsy-proved prostate cancer were examined with a dynamic contrast material-enhanced inversion-prepared dual-contrast gradient-echo sequence (temporal resolution, 1.65 seconds) at 1.5 T to calculate blood volume, interstitial volume, and blood flow. These parameters were correlated with MVD, MVA, and MIA in 95 areas (prostate cancer, n = 36; chronic prostatitis, n = 27; normal prostate tissue, n = 32). For each MR area, five 1-mm(2) squares (original magnification, x100) of the matching histologic area were analyzed. The Wilcoxon signed-rank test was used for statistical analysis. RESULTS: Blood volume correlated poorly with MVD (Spearman correlation coefficient, 0.252; P = .014) but did not correlate at all with MVA (P = .759). Interstitial volume did not correlate with MIA (P = .507). Blood volume differed between patients with prostate cancer and those with a normal prostate (1.49% vs 0.84%, respectively; P < .001). Interstitial volume differed between patients with chronic prostatitis and those with a normal prostate (39.00% vs 22.59%, respectively; P = .022). Blood flow differed between patients with prostate cancer and those with a normal prostate (0.97 mL/[cm(3) x min(-1)] vs 0.34 mL/[cm(3) x min(-1)], respectively; P < .001), between patients with prostate cancer and those with chronic prostatitis (0.97 mL/[cm(3) x min(-1)] vs 0.60 mL/[cm(3) x min(-1)], respectively; P = .026), and between patients with chronic prostatitis and those with a normal prostate (0.60 mL/[cm(3) x min(-1)] vs 0.34 mL/[cm(3) x min(-1)], respectively; P = .023). CONCLUSION: Blood volume and interstitial volume did not reliably correlate with the histologic parameters. Only blood flow enabled differentiation of prostate cancer, chronic prostatitis, and normal prostate tissue. PMID- 19561253 TI - Effects of adaptive section collimation on patient radiation dose in multisection spiral CT. AB - PURPOSE: To evaluate the potential effectiveness of adaptive collimation in reducing computed tomographic (CT) radiation dose owing to z-overscanning by using dose measurements and Monte Carlo (MC) dose simulations. MATERIALS AND METHODS: Institutional review board approval was not necessary. Dose profiles were measured with thermoluminescent dosimeters in CT dose index phantoms and in an Alderson-Rando phantom without and with adaptive section collimation for spiral cardiac and chest CT protocols and were compared with the MC simulated dose profiles. Additional dose measurements were performed with an ionization chamber for scan ranges of 5-50 cm and pitch factors of 0.5-1.5. RESULTS: The measured and simulated dose profiles agreed to within 3%. By using adaptive section collimation, a substantial dose reduction of up to 10% was achieved for cardiac and chest CT when measurements were performed free in air and of 7% on average when measurements were performed in phantoms. For scan ranges smaller than 12 cm, ionization chamber measurements and simulations indicated a dose reduction of up to 38%. CONCLUSION: Adaptive section collimation allows substantial reduction of unnecessary exposure owing to z-overscanning in spiral CT. It can be combined in synergy with other means of dose reduction, such as spectral optimization and automatic exposure control. PMID- 19561254 TI - Rotator cuff calcific tendonitis: short-term and 10-year outcomes after two needle us-guided percutaneous treatment--nonrandomized controlled trial. AB - PURPOSE: To compare short- and long-term outcomes of patients with rotator cuff calcific tendonitis who did and did not undergo ultrasonographically (US)-guided percutaneous treatment. MATERIALS AND METHODS: Institutional review board approval and informed patient consent were obtained. Of patients referred for US guided treatment of rotator cuff calcific tendonitis, 219 (86 men, 133 women; mean age, 40.3 years +/- 10.9 [standard deviation]) were treated; 68 (31 men, 37 women; mean age, 40.2 years +/- 11.3) patients refused treatment and served as control subjects. After local anesthesia was induced, two 16-gauge needles were inserted into the calcific deposit. Saline solution was injected through one needle, and the dissolved calcium was extracted through the other needle. Shoulder joint function was assessed by using Constant scores, and pain was assessed by using visual analogue scale (VAS) scores. Mann-Whitney U and chi(2) tests were performed. RESULTS: At baseline, no significant difference in age or sex distribution, Constant score, or VAS score was detected between treated and nontreated (control) patients. Compared with control subjects, treated patients reported a significant decrease in symptoms at 1 month (mean Constant score, 73.2 +/- 6.2 vs 57.5 +/- 3.9; mean VAS score, 4.8 +/- 0.6 vs 9.1 +/- 0.5), 3 months (mean Constant score, 90.2 +/- 2.6 vs 62.6 +/- 7.2; mean VAS score, 3.3 +/- 0.4 vs 7.3 +/- 1.8), and 1 year (mean Constant score, 91.7 +/- 3.1 vs 78.4 +/- 9.5; mean VAS score, 2.7 +/- 0.5 vs 4.5 +/- 0.9) (P < .001). Symptom scores were not significantly different between the groups at 5 years (mean Constant score, 90.9 +/- 3.6 vs 90.5 +/- 4.8; mean VAS score, 2.6 +/- 0.5 vs 2.8 +/- 0.7) (P >or= .795) and 10 years (mean Constant score, 91.8 +/- 5.0 vs 91.3 +/- 9.6; mean VAS score, 2.5 +/- 0.6 vs 2.7 +/- 0.6) (P >or= .413). CONCLUSION: US-guided percutaneous treatment facilitated prompt shoulder function recovery and pain relief. Treated patients had better outcomes than did nontreated patients at 1 year. However, 5 and 10 years after the procedure, the nontreated group reported outcomes similar to those of the treated group. PMID- 19561255 TI - Age-related iron deposition in the basal ganglia: quantitative analysis in healthy subjects. AB - PURPOSE: To determine the values of iron accumulation in the basal ganglia of healthy volunteers of different ages with R2* and raw signal intensity measurements from T1-weighted magnetic resonance (MR) images, supported by voxel based relaxometry (VBR), and to compare them with previously reported iron concentrations found in autopsy material. MATERIALS AND METHODS: The ethics committee approved the study, and the participants or their parents gave written informed consent. Eighty subjects (41 female and 39 male subjects; age range, 1 80 years) were examined at 1.5 T. For each subject, R2* values were calculated. Curves for R2* versus age were obtained for globus pallidus (GP), putamen, caudate nucleus, substantia nigra (SN), and frontal white matter (FWM). To highlight possible differences in iron concentration among the age decades, VBR was applied. Signal intensity values were estimated on T1-weighted fast low-angle shot images, and regions of interest were drawn in each nucleus. R2* values were also compared with iron concentrations reported in a postmortem study. Statistical analysis was performed (t test), and a difference with P < .05 (FDR corrected) was significant. RESULTS: The curves for R2* versus age showed an exponential increase with increasing age in all the basal ganglia. VBR demonstrated significant differences (P < .05, corrected) in the comparison between the 2nd and the following decades for lenticular nuclei. Good correlation coefficients were found for GP (R(2) = 0.64), putamen (R(2) = 0.51), and SN (R(2) = 0.53) when compared with findings in the postmortem study. Signal intensity curves were similar to the R2* curves. CONCLUSION: R2* measurements can be used to quantify brain iron accumulation and thus may allow better evaluation of neurodegenerative diseases associated with iron deposition. PMID- 19561256 TI - Recurrent glioblastoma multiforme: ADC histogram analysis predicts response to bevacizumab treatment. AB - PURPOSE: To determine if apparent diffusion coefficient (ADC) histogram analysis can stratify progression-free survival in patients with recurrent glioblastoma multiforme (GBM) prior to bevacizumab treatment. MATERIALS AND METHODS: The study was approved by the institutional review board and was HIPAA compliant; informed consent was obtained. Bevacizumab-treated and control patients (41 per cohort) diagnosed with recurrent GBM were analyzed by using whole enhancing tumor ADC histograms with a two normal distribution mixture fitting curve on baseline (pretreatment) magnetic resonance (MR) images to generate ADC classifiers, including the overall mean ADC as well as the mean ADC from the lower curve (ADC(L)). Overall and 6-month progression-free survival (as defined by the Macdonald criteria) was determined by using Cox proportional hazard ratios and the Kaplan-Meier method with log-rank test. RESULTS: For bevacizumab-treated patients, the hazard ratio for progression by 6 months in patients with less than versus greater than mean ADC(L) was 4.1 (95% confidence interval: 1.6, 10.4), and there was a 2.75-fold reduction in the median time to progression. For the control patients, there was no significant difference in median time to progression for the patients with low versus high ADC(L) (hazard ratio, 1.8; 95% confidence interval: 0.9, 3.7). For bevacizumab-treated patients, pretreatment ADC more accurately stratified 6-month progression-free survival than did change in enhancing tumor volume at first follow-up (73% vs 58% accuracy, P = .034). CONCLUSION: Pretreatment ADC histogram analysis can stratify progression-free survival in bevacizumab-treated patients with recurrent GBM. PMID- 19561257 TI - T2 at MR imaging is an objective quantitative measure of cerebral white matter signal intensity abnormality in preterm infants at term-equivalent age. AB - PURPOSE: To compare quantitative T2 relaxometry of cerebral white matter (WM) with qualitative assessment of conventional T2-weighted magnetic resonance (MR) images, to assess the relationship between cerebral WM T2 and region-specific apparent diffusion coefficient (ADC), and to examine WM T2 regional variation in preterm infants at term. MATERIALS AND METHODS: The local ethical committee granted ethical permission for this study; informed parental consent was obtained for each infant. Sixty-two preterm infants born at less than 32 weeks gestation and nine control infants were examined at 1.5 T; T2-weighted fast spin-echo MR images, T2 relaxometry data, and diffusion-weighted MR images were acquired. Conventional T2-weighted MR images were assessed by a pediatric neuroradiologist for diffuse excessive high signal intensity (DEHSI) in WM. Regions of interest were positioned in frontal WM, central WM, and posterior WM at the level of the centrum semiovale. RESULTS: In preterm infants at term, T2 was longer in all WM regions than in control infants; in infants with DEHSI, T2 was longer than in infants without DEHSI and control infants, with posterior WM T2 being longer than central or frontal WM T2. In control infants, T2 was similar in all WM regions. Frontal and posterior WM ADCs were higher in preterm infants at term than in control infants. CONCLUSION: Cerebral WM T2 is an objective quantitative measurement that can easily and rapidly be obtained during clinical MR imaging in preterm infants at term. PMID- 19561258 TI - Feasibility of ex vivo FDG PET of the colon. AB - To facilitate future direct correlations between fluorine 18 fluorodeoxyglucose (FDG)-avid colonic lesions and immunohistochemical assay findings, the authors tested the feasibility of ex vivo FDG positron emission tomography (PET) of the colon resected from humans. In this institutional review board-approved, HIPAA compliant study, the authors, after obtaining informed patient consent, injected FDG intraoperatively in five patients with neoplasms and imaged their resected colons approximately 3 hours later. The colon could be imaged during this fairly limited time interval, and polyps and cancers could be identified. No biologic tissue degradation occurred. The authors concluded that ex vivo FDG PET of the colon is feasible and, when combined with careful histologic and immunohistochemical analyses, may serve as a research tool to determine the mechanisms of the normal colonic uptake of FDG and the localization of FDG in polyps and cancers. PMID- 19561259 TI - Urethrography in men: conventional technique versus clamp method. AB - PURPOSE: To compare examination adequacy and patient discomfort during retrograde urethrography (RUG) performed by using the conventional balloon method versus those of RUG and voiding cystourethrography (VCUG) performed with the clamp method of using drip infusion for the administration of contrast material. MATERIALS AND METHODS: This prospective study was approved by the institutional review board; written informed consent was obtained from all patients. Eighty men (mean age, 64.3 years +/- 16 [standard deviation]; range, 18-85 years) suspected of having urethral stenosis were randomly distributed into two groups for urethrography: a control group (n = 36) and a clamp group (n = 44). In 11 of the 36 patients in the control group, the conventional balloon method could not be used, so these patients were transferred to the clamp group. Drip infusion was used to administer contrast material for RUG, and, except in cases where a suprapubic catheter was used (n = 8), for VCUG. The pain levels reported by patients were recorded by using a verbal descriptor scale (VDS) and a visual analogue scale (VAS). RESULTS: In the control group, RUG was successfully performed in 69% of patients (25 of 36), and mean pain levels recorded on inflation of the balloon were distressing according to the VDS and 4.8 +/- 1.4 (range, 2.3-7.5) according to the VAS. In the clamp group, RUG was successfully performed in all cases; in 69% of patients in this group (38 of 55), the pain level recorded at external compression was no pain according to the VDS and 0 according to the VAS, while mean values in the remaining 31% of patients (17 of 55) were mild pain on the VDS and 0.6 +/- 0.3 (range, 0.3-1.2) on the VAS. Bladder filling for VCUG was achieved with drip infusion in 96% of patients (69 of 72) in an average time of 11 minutes. CONCLUSION: The conventional balloon method of performing RUG is painful and, in some cases, not effective. The clamp method is a simple, well-tolerated procedure that allowed diagnostic evaluation in all cases. Drip infusion enables RUG and VCUG to be performed without the need for syringes or bladder catheters, thus increasing patient comfort. PMID- 19561260 TI - Evaluating variability in tumor measurements from same-day repeat CT scans of patients with non-small cell lung cancer. AB - PURPOSE: To evaluate the variability of tumor unidimensional, bidimensional, and volumetric measurements on same-day repeat computed tomographic (CT) scans in patients with non-small cell lung cancer. MATERIALS AND METHODS: This HIPAA compliant study was approved by the institutional review board, with informed patient consent. Thirty-two patients with non-small cell lung cancer, each of whom underwent two CT scans of the chest within 15 minutes by using the same imaging protocol, were included in this study. Three radiologists independently measured the two greatest diameters of each lesion on both scans and, during another session, measured the same tumors on the first scan. In a separate analysis, computer software was applied to assist in the calculation of the two greatest diameters and the volume of each lesion on both scans. Concordance correlation coefficients (CCCs) and Bland-Altman plots were used to assess the agreements between the measurements of the two repeat scans (reproducibility) and between the two repeat readings of the same scan (repeatability). RESULTS: The reproducibility and repeatability of the three radiologists' measurements were high (all CCCs, >or=0.96). The reproducibility of the computer-aided measurements was even higher (all CCCs, 1.00). The 95% limits of agreements for the computer aided unidimensional, bidimensional, and volumetric measurements on two repeat scans were (-7.3%, 6.2%), (-17.6%, 19.8%), and (-12.1%, 13.4%), respectively. CONCLUSION: Chest CT scans are well reproducible. Changes in unidimensional lesion size of 8% or greater exceed the measurement variability of the computer method and can be considered significant when estimating the outcome of therapy in a patient. PMID- 19561261 TI - Use of a computer-aided detection system to detect missed lung cancer at chest radiography. AB - PURPOSE: To study the ability of a computer-aided detection (CAD) system to detect lung cancer overlooked at initial interpretation by the radiologist. MATERIALS AND METHODS: Institutional review board approval was given for this study. Patient consent was not required; a HIPAA waiver was granted because of the retrospective nature of the data collection. In patients with lung cancer diagnosed from 1995 to 2006 at two institutions, each chest radiograph obtained prior to tumor discovery was evaluated by two radiologists for an overlooked lesion. The size and location of the nodules were documented and graded for subtlety (grades 1-4, 1 = very subtle). Each radiograph with a missed lesion was analyzed by a commercial CAD system, as was the follow-up image at diagnosis. An age- and sex-matched control group was used to assess CAD false-positive rates. RESULTS: Missed lung cancer was found in 89 patients (age range, 51-86 years; mean age, 65 years; 80 men, nine women) on 114 radiographs. Lesion size ranged from 0.4 to 5.5 cm (mean, 1.8 cm). Lesions were most commonly peripheral (n = 63, 71%) and in upper lobes (n = 67, 75%). Lesion subtlety score was 1, 2, 3, or 4 on 43, 49, 17, and five radiographs, respectively. CAD identified 53 (47%) and 46 (52%) undetected lesions on a per-image and per-patient basis, respectively. The average size of lesions detected with CAD was 1.73 cm compared with 1.85 cm for lesions that were undetected (P = .47). A significant difference (P = .017) was found in the average subtlety score between detected lesions (score, 2.06) and undetected lesions (score, 1.68). An average of 3.9 false-positive results occurred per radiograph; an average of 2.4 false-positive results occurred per radiograph for the control group. CONCLUSION: CAD has the potential to detect approximately half of the lesions overlooked by human readers at chest radiography. PMID- 19561262 TI - Biliary atresia: color doppler US findings in neonates and infants. AB - PURPOSE: To describe color Doppler ultrasonographic (US) findings in livers of neonates with biliary atresia (BA) and to compare them with US findings in livers of neonates with non-BA and control subjects. MATERIALS AND METHODS: Institutional review board approval was obtained; acquisition of informed consent was exempted. US and color Doppler US findings were retrospectively reviewed in 64 patients with neonatal cholestasis and 19 control subjects. BA and non-BA were confirmed in 29 and 35 patients, respectively. Three pediatric radiologists assessed US and color Doppler US images, independently documented their findings, and resolved discrepancies by consensus. Triangular cord (TC) sign, gallbladder length, and hepatic artery and portal vein diameters were evaluated on US images. The presence of hepatic subcapsular flow was evaluated on color Doppler US images. Diagnostic value of TC sign and hepatic subcapsular flow in the diagnosis of BA were evaluated. Significance of hepatic artery and portal vein diameters in each group was assessed. RESULTS: In the diagnosis of BA, sensitivity and specificity of the TC sign on US images were 62% and 100%, respectively. On color Doppler US images, hepatic subcapsular flow was detected in all patients with BA and in five patients with non-BA. At the first review, there was a discrepancy between radiologists in interpretation of hepatic subcapsular flow in patients with non-BA. However, consensus was reached at the second review. There was no hepatic subcapsular flow in control subjects. Sensitivity and specificity of hepatic subcapsular flow on color Doppler US images were 100% and 80%-86%, respectively, on the basis of individual interpretations of reviewers. Sensitivity and specificity of hepatic subcapsular flow on color Doppler US images were 100% and 86%, respectively, on the basis of consensus reading. Mean diameter of the hepatic artery in patients with BA (2.1 mm +/- 0.7 [standard deviation]) was significantly larger than that in patients with non-BA (1.5 mm +/ 0.4, P < .001) and control subjects (1.5 mm +/- 0.4, P = .001). CONCLUSION: The presence of hepatic subcapsular flow is useful for differentiating between BA and other causes of neonatal jaundice. PMID- 19561263 TI - High-dose immunoembolization: survival benefit in patients with hepatic metastases from uveal melanoma. AB - PURPOSE: To retrospectively evaluate prognostic factors for survival in patients with uveal melanoma who received chemoembolization (CE) with 1,3-bis (2 chloroethyl)-1-nitrosourea or immunoembolization (IE) with granulocyte-macrophage colony-stimulating factor (GM-CSF) for hepatic metastases. MATERIALS AND METHODS: Fifty-three consecutive patients with uveal melanoma were treated by using CE or IE in clinical trials approved by the Institutional Review Board. Prognostic factors associated with overall survival (OS) and progression-free survival (PFS) in the liver and extrahepatic (systemic) organs were retrospectively evaluated. Covariates of age, sex, preexisting extrahepatic metastases, liver enzyme levels, tumor volume, radiologic response in hepatic metastases, and treatment type were analyzed. RESULTS: Compared with CE, high-dose (>or=1500 microg of GM-CSF) IE resulted in significantly better OS (20.4 vs 9.8 months, P = .005) and systemic PFS (12.4 vs 4.8 months, P = .001) at univariate analysis. Overall, women outlived men (14.4 vs 9.8 months, P = .01). Patients who achieved regression of hepatic metastases after embolization lived much longer than did those who did not achieve regression (27.2 vs 9.9 months, P < .001). At multivariate analysis, prolonged OS was confirmed for women, patients who underwent high-dose IE, younger patients (age < 60 years), and patients with regression of hepatic metastases. Independent predictors of longer systemic PFS included high-dose IE, younger age, and regression of hepatic metastases. No covariate predicted liver PFS except for hepatic response. CONCLUSION: Treatment with high-dose IE prolonged survival of patients with uveal melanoma who received embolization of hepatic metastases and possibly delayed progression of extrahepatic metastases. PMID- 19561264 TI - 64-Detector CT angiography in renal artery stent evaluation: prospective comparison with selective catheter angiography. AB - PURPOSE: To prospectively assess the diagnostic accuracy of 64-detector computed tomographic (CT) renal artery (RA) angiography for the evaluation of RA in-stent restenosis (ISR) by using selective catheter RA angiography as the reference standard. MATERIALS AND METHODS: Institutional review board approval and written informed consent were obtained. Eighty-six patients (46 men, 40 women; mean age +/- standard deviation, 71 years +/- 9) with 95 RA stents were examined with 64 detector CT RA angiography by experienced radiologists 1 day before selective catheter RA angiography was performed by experienced cardiologists. Each group was blinded to the results of the other imaging method. For image reconstruction, a standard (B25f) and an edge-enhancing (B46f) kernel were chosen. An ISR of more than 50% was considered as hemodynamically significant. Sensitivity, specificity, and positive and negative predictive values for the detection of ISR by using CT RA angiography were calculated. RESULTS: At CT RA angiography, 93 (98%) of 95 stents were assessable for diagnosis. Two stents could not be evaluated owing to hardening artifacts affected by vessel calcifications. All nine cases of significant ISR seen at selective catheter RA angiography were correctly diagnosed by using CT RA angiography, giving a sensitivity of 100% and a negative predictive value of 100%. One case of nonsignificant ISR seen at selective catheter RA angiography was interpreted as significant by using CT RA angiography, giving a specificity of 99% and a positive predictive value of 90%. In four of 78 patients without ISR seen at selective catheter RA angiography, CT RA angiography showed nonsignificant ISR, giving a specificity of 95% and a positive predictive value of 56%. CONCLUSION: Sixty-four-detector CT RA angiography can provide an excellent noninvasive technique to help detect and evaluate ISR within the RA stents used in our study. PMID- 19561266 TI - Case 147: langerhans cell histiocytosis of the femur. PMID- 19561267 TI - Comments on "Distribution of renal tumor growth rates determined by using serial volumetric CT measurements". PMID- 19561268 TI - Effective dose to dose-length product coefficients for calculation of CT effective dose. PMID- 19561269 TI - Comment on meta-analyses of trials comparing contrast media. PMID- 19561270 TI - Chronic pancreatitis and diffusion-weighted MR imaging. PMID- 19561271 TI - Expression of concern from the editor. PMID- 19561274 TI - Domiciliary non-invasive ventilation in stable COPD? PMID- 19561275 TI - Biomarkers and community-acquired pneumonia. PMID- 19561276 TI - Breast feeding and childhood asthma. PMID- 19561277 TI - Nutritional effects on asthma aetiology and progression. PMID- 19561279 TI - Beware the pregnant woman with breathlessness. PMID- 19561280 TI - Breast feeding, parental allergy and asthma in children followed for 8 years. The PIAMA birth cohort study. AB - BACKGROUND: It is unclear how the association between breast feeding and asthma develops with age of the child and how this association over time is influenced by maternal or paternal allergy. These factors--the age of the child and maternal or paternal allergy--might partly explain the conflicting results observed in cross-sectional studies. METHODS: The study population consisted of 3115 Dutch children born in 1996/1997 who participated in the PIAMA (Prevention and Incidence of Asthma and Mite Allergy) birth cohort study. Data on breast feeding and asthma (based on wheeze, dyspnoea and prescription of inhaled steroids) were collected by yearly questionnaires. At 8 years, specific immunoglobulin E (IgE) to airborne allergens and bronchial responsiveness were measured. Data were analysed by logistic regression and generalised estimating equations (GEEs), and stratified by maternal and paternal allergic status. RESULTS: 35% (n = 1081) of the children were breast fed for >16 weeks. At 8 years of age, 12.6% (n = 392) had asthma. Breast feeding (>16 weeks vs no breast feeding) was significantly associated with a lower asthma prevalence from 3 to 8 years of age, in children of both non-allergic and allergic mothers. The inverse association between breast feeding and sensitisation to airborne allergens at 8 years was non-significant. Breast feeding was not associated with bronchial hyper-responsiveness. No interaction between breast feeding and gender, maternal allergy or paternal allergy was observed in any of the associations. CONCLUSIONS: Breast feeding is associated with a lower asthma risk in children until 8 years of age without evidence of attenuation and regardless of the family history of allergy. PMID- 19561283 TI - Systemic inflammation: a key factor in the pathogenesis of cardiovascular complications in obstructive sleep apnoea syndrome? AB - Obstructive sleep apnoea syndrome (OSAS) is a highly prevalent disease and is recognised as a major public health burden. Large-scale epidemiological studies have demonstrated an independent relationship between OSAS and various cardiovascular disorders. The pathogenesis of cardiovascular complications in OSAS is not completely understood but a multifactorial aetiology is likely. Inflammatory processes have emerged as critical in the pathogenesis of atherosclerosis at all stages of atheroma formation. Increased levels of various circulating markers of inflammation including tumour necrosis factor alpha (TNFalpha), interleukin 6 (IL6), IL-8 and C-reactive protein (CRP) have been reported as associated with future cardiovascular risk. There is increasing evidence of elevated inflammatory markers in OSAS with a significant fall after effective treatment with continuous positive airway pressure. This evidence is particularly strong for TNFalpha, whereas studies on IL6 and CRP have yielded conflicting results possibly due to the confounding effects of obesity. Cell culture and animal studies have significantly contributed to our understanding of the underlying mechanisms of the association between OSAS and inflammation. Intermittent hypoxia, the hallmark of OSAS, results in activation of pro inflammatory transcription factors such as nuclear factor kappa B (NF-kappaB) and activator protein (AP)-1. These promote activation of various inflammatory cells, particularly lymphocytes and monocytes, with the downstream consequence of expression of pro-inflammatory mediators that may lead to endothelial dysfunction. This review provides a critical analysis of the current evidence for an association between OSAS, inflammation and cardiovascular disease, discusses basic mechanisms that may be responsible for this association and proposes future research possibilities. PMID- 19561284 TI - A case of small cell lung cancer treated with chemoradiotherapy followed by photodynamic therapy. AB - Here, we present the case of a 51-year-old man with limited-stage small cell lung cancer (LS-SCLC) who received concurrent chemoradiotherapy and photodynamic therapy (PDT). The patient was diagnosed as having LS-SCLC with an endobronchial mass in the left main bronchus. Following concurrent chemoradiotherapy, a mass remaining in the left lingular division was treated with PDT. Clinical and histological data indicate that the patient has remained in complete response for 2 years without further treatment. This patient represents a rare case of complete response in LS-SCLC treated with PDT. PMID- 19561286 TI - Obesity in patients with COPD, an undervalued problem? PMID- 19561287 TI - Validation of two activity monitors in patients with COPD. PMID- 19561288 TI - Do all occupational respiratory sensitisers follow the united airways disease model? PMID- 19561289 TI - Epoprostenol use in a National Pulmonary Hypertension Centre from 1997 to 2007. PMID- 19561290 TI - A woman in her mid 30s with a dry cough and breathlessness. PMID- 19561291 TI - Anthracyclines in early-stage breast cancer: is it the end of an era? AB - Anthracycline regimens have been the mainstay of adjuvant care in breast cancer for >20 years. A growing body of clinical experience has uncovered an unacceptable rate of significant cardiac and leukomogenic toxicities. A systematic review of the literature was performed highlighting anthracycline- and nonanthracycline-based adjuvant regimens. The published data suggest that nonanthracycline alternatives are less toxic than anthracycline-containing regimens and equally, if not more, efficacious. Molecular predictors, such as human epidermal growth factor receptor 2 and topoisomerase II alpha, are further refining the optimal role of anthracyclines. With these new advances, the current role of anthracycline-based chemotherapy in early-stage breast cancer demands re examination. PMID- 19561292 TI - Role of hematopoietic stem cell transplant in the management of follicular lymphoma. AB - Despite decades of published data regarding the application of autologous and allogeneic stem cell transplant in patients with follicular lymphoma, there remain no uniform indications for its use in this disease. Autologous transplant has been shown to lead to longer progression-free survival times in randomized trials when compared with postremission interferon-based chemoimmunotherapy. However, the development of rituximab and its use in frontline, salvage, and maintenance therapy complicates the decision to pursue autologous transplant, a modality developed prior to the advent of anti-CD20 monoclonal antibodies. Allogeneic transplant offers the advantages of lymphoma-free grafts and the immunologic graft-versus-lymphoma effect. These factors may confer the possibility of long-term remission, though historically they have been accompanied by high rates of upfront morbidity and mortality, especially in heavily pretreated patients with a poor performance status or chemotherapy refractory disease. Advances in patient selection, human leukocyte antigen (HLA) matching, conditioning regimens, and supportive care have reduced transplant related mortality and the incidence of graft-versus-host disease. Recently published data focus on the incorporation of rituximab and radioimmunoconjugates prior to, during, and following autologous transplant. Furthermore, reduced intensity allogeneic stem cell transplantation has increasingly been used for relapsed follicular lymphoma patients with comorbidities or advanced age. Several recent reports suggest that reduced-intensity regimens may provide a high likelihood of long-term disease-free survival for patients up to 70 years of age with a good performance status, chemotherapy-sensitive disease, and HLA-matched sibling donors. Such patients with relapsed disease should be referred to a transplant center that can enroll them in one of the forthcoming clinical trials that aim to confirm these outcomes. PMID- 19561293 TI - Dominant mutations in the tyrosyl-tRNA synthetase gene recapitulate in Drosophila features of human Charcot-Marie-Tooth neuropathy. AB - Dominant-intermediate Charcot-Marie-Tooth neuropathy (DI-CMT) is characterized by axonal degeneration and demyelination of peripheral motor and sensory neurons. Three dominant mutations in the YARS gene, encoding tyrosyl-tRNA synthetase (TyrRS), have so far been associated with DI-CMT type C. The molecular mechanisms through which mutations in YARS lead to peripheral neuropathy are currently unknown, and animal models for DI-CMTC are not yet available. Here, we report the generation of a Drosophila model of DI-CMTC: expression of the 3 mutant--but not wild type--TyrRS in Drosophila recapitulates several hallmarks of the human disease, including a progressive deficit in motor performance, electrophysiological evidence of neuronal dysfunction and morphological signs of axonal degeneration. Not only ubiquitous, but also neuron-specific expression of mutant TyrRS, induces these phenotypes, indicating that the mutant enzyme has cell-autonomous effects in neurons. Furthermore, biochemical and genetic complementation experiments revealed that loss of enzymatic activity is not a common feature of DI-CMTC-associated mutations. Thus, the DI-CMTC phenotype is not due to haploinsufficiency of aminoacylation activity, but most likely to a gain-of-function alteration of the mutant TyrRS or interference with an unknown function of the WT protein. Our results also suggest that the molecular pathways leading to mutant TyrRS-associated neurodegeneration are conserved from flies to humans. PMID- 19561294 TI - Scaling of strength and lifetime probability distributions of quasibrittle structures based on atomistic fracture mechanics. AB - The failure probability of engineering structures such as aircraft, bridges, dams, nuclear structures, and ships, as well as microelectronic components and medical implants, must be kept extremely low, typically <10(-6). The safety factors needed to ensure it have so far been assessed empirically. For perfectly ductile and perfectly brittle structures, the empirical approach is sufficient because the cumulative distribution function (cdf) of random material strength is known and fixed. However, such an approach is insufficient for structures consisting of quasibrittle materials, which are brittle materials with inhomogeneities that are not negligible compared with the structure size. The reason is that the strength cdf of quasibrittle structure varies from Gaussian to Weibullian as the structure size increases. In this article, a recently proposed theory for the strength cdf of quasibrittle structure is refined by deriving it from fracture mechanics of nanocracks propagating by small, activation-energy controlled, random jumps through the atomic lattice. This refinement also provides a plausible physical justification of the power law for subcritical creep crack growth, hitherto considered empirical. The theory is further extended to predict the cdf of structural lifetime at constant load, which is shown to be size- and geometry-dependent. The size effects on structure strength and lifetime are shown to be related and the latter to be much stronger. The theory fits previously unexplained deviations of experimental strength and lifetime histograms from the Weibull distribution. Finally, a boundary layer method for numerical calculation of the cdf of structural strength and lifetime is outlined. PMID- 19561295 TI - Using adult mosquitoes to transfer insecticides to Aedes aegypti larval habitats. AB - Vector control is a key means of combating mosquito-borne diseases and the only tool available for tackling the transmission of dengue, a disease for which no vaccine, prophylaxis, or therapeutant currently exists. The most effective mosquito control methods include a variety of insecticidal tools that target adults or juveniles. Their successful implementation depends on impacting the largest proportion of the vector population possible. We demonstrate a control strategy that dramatically improves the efficiency with which high coverage of aquatic mosquito habitats can be achieved. The method exploits adult mosquitoes as vehicles of insecticide transfer by harnessing their fundamental behaviors to disseminate a juvenile hormone analogue (JHA) between resting and oviposition sites. A series of field trials undertaken in an Amazon city (Iquitos, Peru) showed that the placement of JHA dissemination stations in just 3-5% of the available resting area resulted in almost complete coverage of sentinel aquatic habitats. More than control mortality occurred in 95-100% of the larval cohorts of Aedes aegypti developing at those sites. Overall reductions in adult emergence of 42-98% were achieved during the trials. A deterministic simulation model predicts amplifications in coverage consistent with our observations and highlights the importance of the residual activity of the insecticide for this technique. PMID- 19561296 TI - Chemistry of fast electrons. AB - A chemicurrent is a flux of fast (kinetic energy approximately > 0.5-1.3 eV) metal electrons caused by moderately exothermic (1-3 eV) chemical reactions over high work function (4-6 eV) metal surfaces. In this report, the relation between chemicurrent and surface chemistry is elucidated with a combination of top-down phenomenology and bottom-up atomic-scale modeling. Examination of catalytic CO oxidation, an example which exhibits a chemicurrent, reveals 3 constituents of this relation: The localization of some conduction electrons to the surface via a reduction reaction, 0.5 O(2) + deltae(-) --> O(delta(-)) (Red); the delocalization of some surface electrons into a conduction band in an oxidation reaction, O(delta(-)) + CO --> CO(2)(delta-) --> CO(2) + deltae(-) (Ox); and relaxation without charge transfer (Rel). Juxtaposition of Red, Ox, and Rel produces a daunting variety of metal electronic excitations, but only those that originate from CO(2) reactive desorption are long-range and fast enough to dominate the chemicurrent. The chemicurrent yield depends on the universality class of the desorption process and the distribution of the desorption thresholds. This analysis implies a power-law relation with exponent 2.66 between the chemicurrent and the heat of adsorption, which is consistent with experimental findings for a range of systems. This picture also applies to other oxidation-reduction reactions over high work function metal surfaces. PMID- 19561297 TI - Genomic-anatomic evidence for distinct functional domains in hippocampal field CA1. AB - Functional heterogeneity has been investigated for decades in the hippocampal region of the mammalian cerebral cortex, and evidence for vaguely defined "dorsal" and "ventral" regions is emerging. Direct evidence that hippocampal field CA1 displays clear regional, laminar, and pyramidal neuron differentiation is presented here, based on a systematic high-resolution analysis of a publicly accessible, genome-wide expression digital library (Allen Brain Atlas) [Lein et al. (2007) Genome-wide atlas of gene expression in the adult mouse brain. Nature 445:168-176]. First, genetic markers reveal distinct spatial expression domains and subdomains along the longitudinal (dorsal/septal/posterior to ventral/temporal/anterior) axis of field CA1. Second, genetic markers divide field CA1 pyramidal neurons into multiple subtypes with characteristic laminar distributions. And third, subcortical brain regions receiving axonal projections from molecularly distinct spatial domains of field CA1 display distinct global gene expression patterns, suggesting that field CA1 spatial domains may be genetically wired independently to form distinct functional networks related to cognition and emotion. Insights emerging from this genomic-anatomic approach provide a starting point for a detailed analysis of differential hippocampal structure-function organization. PMID- 19561298 TI - GPR17 is a negative regulator of the cysteinyl leukotriene 1 receptor response to leukotriene D4. AB - The cysteinyl leukotrienes (cys-LTs) are proinflammatory lipid mediators acting on the type 1 cys-LT receptor (CysLT(1)R) to mediate smooth muscle constriction and vascular permeability. GPR17, a G protein-coupled orphan receptor with homology to the P2Y and cys-LT receptors, failed to mediate calcium flux in response to leukotriene (LT) D(4) with stable transfectants. However, in stable cotransfections of 6xHis-tagged GPR17 with Myc-tagged CysLT(1)R, the robust CysLT(1)R-mediated calcium response to LTD(4) was abolished. The membrane expression of the CysLT(1)R analyzed by FACS with anti-Myc Ab was not reduced by the cotransfection, yet both LTD(4)-elicited ERK phosphorylation and the specific binding of [(3)H]LTD(4) to microsomal membranes were fully inhibited. CysLT(1)R and GPR17 expressed in transfected cells were coimmunoprecipitated and identified by Western blots, and confocal immunofluorescence microscopy revealed that GPR17 and CysLT(1)R colocalize on the cell surface of human peripheral blood monocytes. Lentiviral knockdown of GPR17 in mouse bone marrow-derived macrophages (BMMPhis) increased both the membrane expression of CysLT(1)R protein by FACS analysis and the LTD(4)-elicited calcium flux in a dose-dependent manner as compared with control BMMPhis, indicating a negative regulatory function of GPR17 for CysLT(1)R in a primary cell. In IgE-dependent passive cutaneous anaphylaxis, GPR17 deficient mice showed a marked and significant increase in vascular permeability as compared with WT littermates, and this vascular leak was significantly blocked by pretreatment of the mice with the CysLT(1)R antagonist, MK-571. Taken together, our findings suggest that GPR17 is a ligand-independent, constitutive negative regulator for the CysLT(1)R that suppresses CysLT(1)R-mediated function at the cell membrane. PMID- 19561299 TI - Programmed loss of millions of base pairs from a vertebrate genome. AB - In general, the strict preservation of broad-scale structure is thought to be critical for maintaining the precisely tuned functionality of vertebrate genomes, although nearly all vertebrate species undergo a small number of programmed local rearrangements during development (e.g., remodeling of adaptive immune receptor loci). However, a limited number of metazoan species undergo much more extensive reorganizations as a normal feature of their development. Here, we show that the sea lamprey (Petromyzon marinus), a jawless vertebrate, undergoes a dramatic remodeling of its genome, resulting in the elimination of hundreds of millions of base pairs (and at least one transcribed locus) from many somatic cell lineages during embryonic development. These studies reveal the highly dynamic nature of the lamprey genome and provide the first example of broad-scale programmed rearrangement of a definitively vertebrate genome. Understanding the mechanisms by which this vertebrate species regulates such extensive remodeling of its genome will provide invaluable insight into factors that can promote stability and change in vertebrate genomes. PMID- 19561300 TI - Solution structure of the silkworm betaGRP/GNBP3 N-terminal domain reveals the mechanism for beta-1,3-glucan-specific recognition. AB - The beta-1,3-glucan recognition protein (betaGRP)/Gram-negative bacteria-binding protein 3 (GNBP3) is a crucial pattern-recognition receptor that specifically binds beta-1,3-glucan, a component of fungal cell walls. It evokes innate immunity against fungi through activation of the prophenoloxidase (proPO) cascade and Toll pathway in invertebrates. The betaGRP consists of an N-terminal beta-1,3 glucan-recognition domain and a C-terminal glucanase-like domain, with the former reported to be responsible for the proPO cascade activation. This report shows the solution structure of the N-terminal beta-1,3-glucan recognition domain of silkworm betaGRP. Although the N-terminal domain of betaGRP has a beta-sandwich fold, often seen in carbohydrate-binding modules, both NMR titration experiments and mutational analysis showed that betaGRP has a binding mechanism which is distinct from those observed in previously reported carbohydarate-binding domains. Our results suggest that betaGRP is a beta-1,3-glucan-recognition protein that specifically recognizes a triple-helical structure of beta-1,3 glucan. PMID- 19561302 TI - ANO2 is the cilial calcium-activated chloride channel that may mediate olfactory amplification. AB - For vertebrate olfactory signal transduction, a calcium-activated chloride conductance serves as a major amplification step. However, the molecular identity of the olfactory calcium-activated chloride channel (CaCC) is unknown. Here we report a proteomic screen for cilial membrane proteins of mouse olfactory sensory neurons (OSNs) that identified all the known olfactory transduction components as well as Anoctamin 2 (ANO2). Ano2 transcripts were expressed specifically in OSNs in the olfactory epithelium, and ANO2::EGFP fusion protein localized to the OSN cilia when expressed in vivo using an adenoviral vector. Patch-clamp analysis revealed that ANO2, when expressed in HEK-293 cells, forms a CaCC and exhibits channel properties closely resembling the native olfactory CaCC. Considering these findings together, we propose that ANO2 constitutes the olfactory calcium activated chloride channel. PMID- 19561303 TI - Functional implications of multistage copper binding to the prion protein. AB - The prion protein (PrP) is responsible for a group of neurodegenerative diseases called the transmissible spongiform encephalopathies. The normal function of PrP has not yet been discovered, but indirect evidence suggests a linkage to its ability to bind copper. In this article, low-copper-concentration bindings of Cu(2+) to PrP are investigated by using a recently developed hybrid density functional theory (DFT)/DFT method. It is found that at the lowest copper concentrations, the binding site consists of 4 histidine residues coordinating the copper through epsilon imidazole nitrogens. At higher concentrations, 2 histidines are involved in the binding, one of them in the axial position. These results are in good agreement with existing experimental data. Comparison of free energies for all modes of coordination shows that when enough copper is available, the binding sites will spontaneously rearrange to accommodate more copper ions, despite the fact that binding energy per copper ion decreases with concentration. These findings support the hypothesis that PrP acts as a copper buffer in vivo, protecting other proteins from the attachment of copper ions. Using large-scale classical molecular dynamics, we also probe the structure of full-length copper-bound PrP, including its unfolded N-terminal domain. The results show that copper attachment leads to rearrangement of the structure of the Cu-bonded octarepeat region and to development of turns in areas separating copper-bound residues. These turns make the flexible N-terminal domain more rigid and thus more resistant to misfolding. The last result suggests that copper binding plays a beneficial role in the initial stages of prion diseases. PMID- 19561304 TI - Subseafloor sedimentary life in the South Pacific Gyre. AB - The low-productivity South Pacific Gyre (SPG) is Earth's largest oceanic province. Its sediment accumulates extraordinarily slowly (0.1-1 m per million years). This sediment contains a living community that is characterized by very low biomass and very low metabolic activity. At every depth in cored SPG sediment, mean cell abundances are 3 to 4 orders of magnitude lower than at the same depths in all previously explored subseafloor communities. The net rate of respiration by the subseafloor sedimentary community at each SPG site is 1 to 3 orders of magnitude lower than the rates at previously explored sites. Because of the low respiration rates and the thinness of the sediment, interstitial waters are oxic throughout the sediment column in most of this region. Consequently, the sedimentary community of the SPG is predominantly aerobic, unlike previously explored subseafloor communities. Generation of H(2) by radiolysis of water is a significant electron-donor source for this community. The per-cell respiration rates of this community are about 2 orders of magnitude higher (in oxidation/reduction equivalents) than in previously explored anaerobic subseafloor communities. Respiration rates and cell concentrations in subseafloor sediment throughout almost half of the world ocean may approach those in SPG sediment. PMID- 19561305 TI - Genetic and antibody-mediated reprogramming of natural killer cell missing-self recognition in vivo. AB - Natural killer (NK) cells are lymphocytes of the innate immune system able to recognize and kill tumors lacking self-MHC class I molecules. This "missing-self" recognition is mediated by the lack of engagement of MHC class I-specific inhibitory NK cell receptors that include the killer cell Ig-like receptors (KIR) in humans and Ly49 molecules in mice. A promising immunotherapeutic strategy against MHC class I(+) cancer cells is to block NK cell inhibitory receptors using monoclonal antibodies (mAb). However, interactions between MHC class I molecules and their inhibitory receptors are also required for the acquisition of NK cell functional competence, a process referred as to "education." In addition, inhibitory receptors are involved in self-tolerance on educated NK cells. Here, we developed a preclinical mouse model in which all NK cells are educated by a single transgenic inhibitory receptor, human KIR2DL3, through the engagement with its HLA-Cw3 ligand. This approach revealed that NK cells could be reprogrammed to control the development of mouse syngenic tumors in vivo. Moreover, in vivo anti KIR mAb treatment induced the killing of HLA(+) target cells without breaking self-tolerance. Finally, the long-term infusion of anti-KIR mAb neither abolished NK cell education nor tumor cell recognition. Therefore, these results strongly support the use of inhibitory receptor blockade in cancer patients. PMID- 19561307 TI - Novel strategy for measuring creatine kinase reaction rate in the in vivo heart. AB - In the heart, the creatine kinase (CK) system plays an important role in the cascade of ATP production, transportation, and utilization. The forward pseudo first-order rate constant for the CK reaction can be measured noninvasively by the (31)P-magnetic resonance (MR) spectroscopy magnetization saturation transfer (MST) techniques. However, the measurement of MST in the in vivo heart is limited by the lengthy data acquisition time, especially for studies requiring spatial localization. This technical report presents a new method for measuring ATP production rate via CK that can reduce the MST data acquisition time by 82%. This method is validated using an in vivo pig model to evaluate the forward pseudo first-order rate constant of myocardial CK reaction noninvasively. PMID- 19561306 TI - Apolipoprotein A-I mimetic peptide treatment inhibits inflammatory responses and improves survival in septic rats. AB - Systemic inflammation induces a multiple organ dysfunction syndrome that contributes to morbidity and mortality in septic patients. Since increasing plasma apolipoprotein A-I (apoA-I) and HDL may reduce the complications of sepsis, we tested the hypothesis that the apoA-I mimetic peptide 4F confers similar protective effects in rats undergoing cecal ligation and puncture (CLP) injury. Male Sprague-Dawley rats were randomized to undergo CLP or sham surgery. IL-6 levels were significantly elevated in plasma by 6 h after CLP surgery compared with shams. In subsequent studies, CLP rats were further subdivided to receive vehicle or 4F (10 mg/kg) by intraperitoneal injection, 6 h after sepsis induction. Sham-operated rats received saline. Echocardiographic studies showed a reduction in left ventricular end-diastolic volume, stroke volume, and cardiac output (CO) 24 h after CLP surgery. These changes were associated with reduced blood volume and left ventricular filling pressure. 4F treatment improved blood volume status, increased CO, and reduced plasma IL-6 in CLP rats. Total cholesterol (TC) and HDL were 79 +/- 5 and 61 +/- 4 mg/dl, respectively, in sham rats. TC was significantly reduced in CLP rats (54 +/- 3 mg/dl) due to a reduction in HDL (26 +/- 3 mg/dl). 4F administration to CLP rats attenuated the reduction in TC (69 +/- 4 mg/dl) and HDL (41 +/- 3 mg/dl) and prevented sepsis induced changes in HDL protein composition. Increased plasma HDL in 4F-treated CLP rats was associated with an improvement in CO and reduced mortality. It is proposed that protective effects of 4F are related to its ability to prevent the sepsis-induced reduction in plasma HDL. PMID- 19561308 TI - Parent-of-origin effects on cardiac response to pressure overload in mice. AB - Left ventricular (LV) hypertrophy (LVH) is an independent risk factor for cardiovascular mortality and is commonly caused by hypertension. In rodents, transverse aortic constriction (TAC) is a model regularly employed in mechanistic studies of the response of the LV to pressure overload. We previously reported that inbred strains of male mice manifest different cardiac responses to TAC, with C57BL/6J (B6) developing LV dilatation and impaired contractility and 129S1/SvImJ (129) males displaying concentric LVH. In the present study, we investigated sex and parent-of-origin effects on the response to TAC by comparing cardiac function, organ weights, expression of cardiac hypertrophy markers, and histology in female B6 and female 129 mice and in F1 progeny of reciprocal crosses between B6 and 129 mice (B6129F1 and 129B6F1). Five weeks after TAC, heart weight increased to the greatest extent in 129B6F1 mice and the least extent in 129 and B6129F1 mice. Female 129B6F1 and B6 mice were relatively protected from the increase in heart weight that occurs in their male counterparts with pressure overload. The response to TAC in 129 consomic mice bearing the B6 Y chromosome resembled that of 129 rather than 129B6F1 mice, indicating that the B6 Y chromosome does not account for the differences in the reciprocal cross. Our results suggest that susceptibility to LVH is more complex than simple Mendelian inheritance and that parental origin effects strongly impact the LV response to TAC in these commonly used inbred strains. PMID- 19561309 TI - Arrhythmogenic consequences of intracellular calcium waves. AB - Intracellular Ca(2+) (Ca(i)(2+)) waves are known to cause delayed afterdepolarizations (DADs), which have been associated with arrhythmias in cardiac disease states such as heart failure, catecholaminergic polymorphic ventricular tachycardia, and digitalis toxicity. Here we show that, in addition to DADs, Ca(i)(2+) waves also have other consequences relevant to arrhythmogenesis, including subcellular spatially discordant alternans (SDA, in which the amplitude of the local Ca(i)(2+) transient alternates out of phase in different regions of the same cell), sudden repolarization changes promoting the dispersion of refractoriness, and early afterdepolarizations (EADs). Ca(i)(2+) was imaged using a charge-coupled device-based system in fluo-4 AM-loaded isolated rabbit ventricular myocytes paced at constant or incrementally increasing rates, using either field stimulation, current clamp, or action potential (AP) clamp. Ca(i)(2+) waves were induced by Bay K 8644 (50 nM) + isoproterenol (100 nM), or low temperature. When pacing was initiated during a spontaneous Ca(i)(2+) wave, SDA occurred abruptly and persisted during pacing. Similarly, during rapid pacing, SDA typically arose suddenly from spatially concordant alternans, due to an abrupt phase reversal of the subcellular Ca(i)(2+) transient in a region of the myocyte. Ca(i)(2+) waves could be visualized interspersed with AP-triggered Ca(i)(2+) transients, producing a rich variety of subcellular Ca(i)(2+) transient patterns. With free-running APs, complex Ca(i)(2+) release patterns were associated with DADs, EADs, and sudden changes in AP duration. These findings link Ca(i)(2+) waves directly to a variety of arrhythmogenic phenomena relevant to the intact heart. PMID- 19561310 TI - Speckle tracking echocardiography in the assessment of mouse models of cardiac dysfunction. AB - Two-dimensional (2-D) speckle tracking echocardiography (STE) accurately quantifies circumferential strain (S(circ)) and radial strain (S(rad)) in humans and in large and small animals. This study was performed to assess sensitivity of S(circ) and S(rad) to left ventricular (LV) dysfunction in mouse models. We performed 2-D and M-mode echocardiography 1) in 6 mice during superficial and profound isoflurane anesthesia, 2) serially in 12 mice to monitor the development of heart failure induced by transverse aortic constriction (TAC) and in 8 corresponding control mice, and 3) in 26 mice with varying degrees of TAC-induced heart failure and 12 corresponding control mice immediately before euthanasia. Fractional shortening (FS) and LV mass were measured from standard M-mode tracings, whereas S(circ) and S(rad) were derived by STE. Percent fibrosis and myocyte diameters were assessed from whole heart cross-sectional specimens stained by Masson trichrome. Profound isoflurane anesthesia decreased S(circ) (P = 0.027) but not S(rad) (P > 0.05). Mice subjected to TAC showed an immediate and sustained decrease in FS (P = 0.035), S(circ) (P = 0.016), and S(rad) (P = 0.012). S(circ) showed better correlation with FS (r = 0.56 and P < 0.0001) and LV mass (r = 0.42 and P = 0.0003) than S(rad) (r = 0.54 and P < 0.0001 for FS and r = 0.37 and P = 0.014 for LV mass, respectively). Percent fibrosis correlated better with S(circ) (r = 0.46 and P = 0.004) than with S(rad) (r = -0.32 and P = 0.05), whereas myocyte diameter showed similar correlation with both strains (r = 0.45 and r = -0.44, respectively, and P = 0.006 for both). STE correctly identifies LV dysfunction and histological changes in mice and can be used for the serial assessment of cardiac remodeling in murine models. PMID- 19561312 TI - CORM-3-derived CO modulates polymorphonuclear leukocyte migration across the vascular endothelium by reducing levels of cell surface-bound elastase. AB - Recently, it has been shown that carbon monoxide (CO)-releasing molecule (CORM) released CO can suppress inflammation. In this study, we assessed the effects and potential mechanisms of a ruthenium-based water-soluble CO carrier [tricarbonylchloroglycinate-ruthenium(II) (CORM-3)] in the modulation of polymorphonuclear leukocyte (PMN) inflammatory responses in an experimental model of sepsis. Sepsis in mice was induced by cecal ligation and puncture. CORM-3 (3 mg/kg iv) was administered 15 min after the induction of cecal ligation and puncture. PMN accumulation in the lung (myeloperoxidase assay), bronchoalveolar lavage (BAL) fluid, and lung vascular permeability (protein content in BAL fluid) were assessed 6 h later. In in vitro experiments, human PMNs were primed with LPS (10 ng/ml) and subsequently stimulated with formyl-methionyl-leucylphenylalanine (fMLP; 100 nM). PMN production of ROS (L-012/dihydrorhodamine-123 oxidation), degranulation (release of elastase), and PMN rolling, adhesion, and migration to/across human umbilical vein endothelial cells (HUVECs) were assessed in the presence or absence of CORM-3 (1-100 muM). The obtained results indicated that systemically administered CORM-3 attenuates PMN accumulation and vascular permeability in the septic lung. Surprisingly, in in vitro experiments, treatment of PMNs with CORM-3 further augmented LPS/fMLP-induced ROS production and the release of elastase. The latter effects, however, were accompanied by an inability of PMNs to mobilize elastase to the cell surface (plasma membrane), an event required for efficient PMN transendothelial migration. The CORM-3-induced decrease in cell surface levels of elastase was followed by decreased PMN rolling/adhesion to HUVECs and complete prevention of PMN migration across HUVECs. In contrast, treatment of HUVECs with CORM-3 had no effect on PMN transendothelial migration. Taken together, these findings indicate that, in sepsis, CORM3-released CO, while further amplifying ROS production and degranulation of PMNs, concurrently reduces the levels of cell surface-bound elastase, which contributes to suppressed PMN transendothelial migration. PMID- 19561311 TI - Resveratrol blocks interleukin-18-EMMPRIN cross-regulation and smooth muscle cell migration. AB - Vascular smooth muscle cell (SMC) migration is an important mechanism in atherogenesis and postangioplasty arterial remodeling. Previously, we demonstrated that the proinflammatory cytokine interleukin (IL)-18 is a potent inducer of SMC migration. Since extracellular matrix metalloproteinase inducer (EMMPRIN) stimulates ECM degradation and facilitates cell migration, we investigated whether IL-18 and EMMPRIN regulate each other's expression, whether their cross talk induces SMC migration, and whether the phytoalexin resveratrol inhibits IL-18-EMMPRIN signaling and SMC migration. Our studies demonstrate that 1) IL-18 induces EMMPRIN mRNA and protein expressions and stimulates EMMPRIN secretion from human aortic SMCs; 2) IL-18 stimulates EMMPRIN expression via oxidative stress and phosphatidylinositol 3-kinase (PI3K)-Akt-ERK signaling; 3) IL-18-stimulated SMC migration is significantly blunted by EMMPRIN knockdown, EMMPRIN function-blocking antibodies, or adenoviral transduction of mutant EMMPRIN; 4) conversely, EMMPRIN stimulates IL-18 expression and secretion via PI3K, Akt, and ERK; and 5) resveratrol attenuates IL-18- and EMMPRIN-mediated PI3K, Akt, and ERK activations; blunts IL-18-mediated oxidative stress; blocks IL 18-EMMPRIN cross-regulation; and inhibits SMC migration. Collectively, our results demonstrate that the coexpression and regulation of IL-18 and EMMPRIN in the vessel wall may amplify the inflammatory cascade and promote atherosclerosis and remodeling. Resveratrol, via its antioxidant and anti-inflammatory properties, has the potential to inhibit the progression of atherosclerosis by blocking IL-18 and EMMPRIN cross-regulation and SMC migration. PMID- 19561313 TI - Decreased cardiac expression of vascular endothelial growth factor and redox imbalance in murine diabetic cardiomyopathy. AB - Type 1 diabetes is associated with a unique form of cardiomyopathy that is present without atherosclerosis. Redox imbalance and/or changes in vascular endothelial growth factor (VEGF) expression have been associated with diabetes related cardiomyopathy. However, the mechanisms of these changes and their interrelationships remain unclear. Using a murine type 1 diabetes model, we tested the hypothesis that alterations in cardiac performance are associated with decreased cardiac microvascular prevalence, as well as downregulation of VEGF isoforms. We also investigated oxidative stress as a contributor to regulate individual VEGF isoforms and microvascular rarefaction. Significant and rapid hyperglycemia was observed at 1 wk post-streptozotocin (STZ) and persisted throughout the 5-wk study. Left ventricular (LV) fractional shortening was reduced at week 1 and 5 post-STZ insult relative to age-matched controls. We also observed the early reduction in E/A ratio at 1 wk. Immunostaining for CD31 and digital image analysis demonstrated a 35% reduction in microvessels/myocardial area, indicative of rarefaction, which was highly correlated with fractional shortening. Furthermore, a significant increase in the prevalence of protein 3 nitrotyrosine was observed in the diabetic cardiac tissue, which was inversely associated with microvascular rarefaction. The expressions of three VEGF isoforms were significantly reduced to different extents. The reduction of VEGF(164) was associated with GSSG accumulation. These data demonstrate that the mouse model of STZ-induced diabetes has hallmark features observed in humans with respect to nonischemic systolic and diastolic performance and microvascular rarefaction, which are associated with changes in VEGF isoform expression and redox imbalance in the myocardium. PMID- 19561314 TI - Spinal nociceptin mediates electroacupuncture-related modulation of visceral sympathoexcitatory reflex responses in rats. AB - The role of nociceptin and its spinal cord neural pathways in electroacupuncture (EA)-related inhibition of visceral excitatory reflexes is not clear. Nociceptin/orphanin FQ (N/OFQ) is an endogenous ligand for a G protein-coupled receptor, called the N/OFQ peptide (NOP) receptor, which has been found to be distributed in the spinal cord. The present study investigated the importance of this system in visceral-cardiovascular reflex modulation during EA. Cardiovascular pressor reflex responses were induced by gastric distension in Sprague-Dawley rats anesthetized by ketamine and xylazine. An intrathecal injection of nociceptin (10 nM) at T1-2 attenuated the pressor responses by 35%, similar to the influence of EA at P 5-6 (42% decrease). An intrathecal injection of the NOP antagonist, [N-Phe(1)]nociceptin(1-13) NH(2), partially reversed the EA response. Pretreatment with the opioid receptor antagonist naloxone did not alter the EA-like inhibitory effect of nociceptin on the pressor reflex, whereas a combination of nociceptin receptor antagonist with naloxone completely abolished the EA response. An intrathecal injection of nociceptin attenuated the pressor responses to the electrical stimulation of the rostral ventrolateral medulla by 46%, suggesting that nociceptin can regulate sympathetic outflow. Furthermore, a bilateral microinjection of NOP antagonist into either the dorsal horn or the intermediolateral column at T1 partially reversed the EA inhibitory effect. These results suggest that nociceptin in the spinal cord mediates part of the EA-related modulation of visceral reflex responses. PMID- 19561315 TI - Reconstruction of myocardial tissue motion and strain fields from displacement encoded MR imaging. AB - A quantitative analysis of myocardial mechanics is fundamental to understanding cardiac function, diagnosis of heart disease, and assessment of therapeutic intervention. Displacement encoding with stimulated-echo (DENSE) magnetic resonance imaging (MRI) technique was developed to track the three-dimensional (3D) displacement vector of discrete material grid points in the myocardial tissue. Despite the wealth of information gained from DENSE images, the current software only provides two-dimensional in-plane deformation. The objective of this study is to introduce a postprocessing method to reconstruct and visualize continuous dynamic 3D displacement and strain fields in the ventricular wall from DENSE data. An anatomically accurate hexagonal finite-element model of the left ventricle (LV) is reconstructed by fitting a prolate spheroidal primitive to contour points of the epi- and endocardial surfaces. The continuous displacement field in the model is described mathematically based on the discrete DENSE vectors using a minimization method with smoothness regularization. Based on the displacement, heart motion and myocardial stretch (or strain) are analyzed. Illustratory computations were conducted with DENSE data of three infarcted and one normal sheep ventricles. The full 3D results show stronger overall axial shortening, wall thickening, and twisting of the normal LV compared with the infarcted hearts. Local myocardial stretches show a dyskinetic LV in the apical region, dilation of apex in systole, and a compensatory increase in strain in the healthy basal region as a compensatory mechanism. We conclude that the proposed postprocessing method significantly extends the utility of DENSE MRI, which may provide a patient-specific 3D model of cardiac mechanics. PMID- 19561316 TI - Dewey defeats Truman and cancer statistics. PMID- 19561317 TI - Defining determinants of pancreatic cancer risk: are we making progress? PMID- 19561318 TI - Dietary fatty acids and pancreatic cancer in the NIH-AARP diet and health study. AB - BACKGROUND: Previous research relating dietary fat, a modifiable risk factor, to pancreatic cancer has been inconclusive. METHODS: We prospectively analyzed the association between intakes of fat, fat subtypes, and fat food sources and exocrine pancreatic cancer in the National Institutes of Health-AARP Diet and Health Study, a US cohort of 308 736 men and 216 737 women who completed a 124 item food frequency questionnaire in 1995-1996. Hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated using Cox proportional hazards regression models, with adjustment for energy intake, smoking history, body mass index, and diabetes. Statistical tests were two-sided. RESULTS: Over an average follow-up of 6.3 years, 865 men and 472 women were diagnosed with exocrine pancreatic cancer (45.0 and 34.5 cases per 100 000 person-years, respectively). After multivariable adjustment and combination of data for men and women, pancreatic cancer risk was directly related to the intakes of total fat (highest vs lowest quintile, 46.8 vs 33.2 cases per 100 000 person-years, HR = 1.23, 95% CI = 1.03 to 1.46; P(trend) = .03), saturated fat (51.5 vs 33.1 cases per 100 000 person-years, HR = 1.36, 95% CI = 1.14 to 1.62; P(trend) < .001), and monounsaturated fat (46.2 vs 32.9 cases per 100 000 person-years, HR = 1.22, 95% CI = 1.02 to 1.46; P(trend) = .05) but not polyunsaturated fat. The associations were strongest for saturated fat from animal food sources (52.0 vs 32.2 cases per 100 000 person-years, HR = 1.43, 95% CI = 1.20 to 1.70; P(trend) < .001); specifically, intakes from red meat and dairy products were both statistically significantly associated with increased pancreatic cancer risk (HR = 1.27 and 1.19, respectively). CONCLUSION: In this large prospective cohort with a wide range of intakes, dietary fat of animal origin was associated with increased pancreatic cancer risk. PMID- 19561319 TI - Functional analysis of cis- and trans-acting elements of the Candida albicans CDR2 promoter with a novel promoter reporter system. AB - Azole resistance in Candida albicans can be mediated by the upregulation of the ATP binding cassette transporter genes CDR1 and CDR2. Both genes are regulated by a cis-acting element called the drug-responsive element (DRE), with the consensus sequence 5'-CGGAWATCGGATATTTTTTT-3', and the transcription factor Tac1p. In order to analyze in detail the DRE sequence necessary for the regulation of CDR1 and CDR2 and properties of TAC1 alleles, a one-hybrid system was designed. This system is based on a P((CDR2))-HIS3 reporter system in which complementation of histidine auxotrophy can be monitored by activation of the reporter system by CDR2-inducing drugs such as estradiol. Our results show that most of the modifications within the DRE, but especially at the level of CGG triplets, strongly reduce CDR2 expression. The CDR2 DRE was replaced by putative DREs deduced from promoters of coregulated genes (CDR1, RTA3, and IFU5). Surprisingly, even if Tac1p was able to bind these putative DREs, as shown by chromatin immunoprecipitation, those from RTA3 and IFU5 did not functionally replace the CDR2 DRE. The one-hybrid system was also used for the identification of gain-of function (GOF) mutations either in TAC1 alleles from clinical C. albicans isolates or inserted in TAC1 wild-type alleles by random mutagenesis. In all, 17 different GOF mutations were identified at 13 distinct positions. Five of them (G980E, N972D, A736V, T225A, and N977D) have already been described in clinical isolates, and four others (G980W, A736T, N972S, and N972I) occurred at already described positions, thus suggesting that GOF mutations can occur in a limited number of positions in Tac1p. In conclusion, the one-hybrid system developed here is rapid and powerful and can be used for characterization of cis- and trans acting elements in C. albicans. PMID- 19561320 TI - Programming tumor-reactive effector memory CD8+ T cells in vitro obviates the requirement for in vivo vaccination. AB - Naive and memory CD8(+) T cells can undergo programmed activation and expansion in response to a short T-cell receptor stimulus, but the extent to which in vitro programming can qualitatively substitute for an in vivo antigen stimulation remains unknown. We show that self-/tumor-reactive effector memory CD8(+) T cells (T(EM)) programmed in vitro either with peptide-pulsed antigen-presenting cells or plate-bound anti-CD3/anti-CD28 embark on a highly stereotyped response of in vivo clonal expansion and tumor destruction nearly identical to that of vaccine stimulated T(EM) cells. This programmed response was associated with an interval of antigen-independent interferon-gamma (IFN-gamma) release that facilitated the dynamic expression of the major histocompatibility complex class I restriction element H-2D(b) on responding tumor cells, leading to recognition and subsequent tumor lysis. Delaying cell transfer for more than 24 hours after stimulation or infusion of cells deficient in IFN-gamma entirely abrogated the benefit of the programmed response, whereas transfer of cells unable to respond to IFN-gamma had no detriment to antitumor immunity. These findings extend the phenomenon of a programmable effector response to memory CD8(+) T cells and have major implications for the design of current adoptive-cell transfer trials. PMID- 19561321 TI - Lymphopenia-induced spontaneous T-cell proliferation as a cofactor for autoimmune disease development. AB - Lymphopenia is thought to be a major cause of tolerance breakdown. In a lymphopenic environment, self-recognition events induce some T cells to expand strongly (a mechanism known as spontaneous proliferation). In this study, we show that in C57BL/6 mice, the repertoire resulting from lymphopenia-induced spontaneous CD4(+) T-cell proliferation included a proportion of regulatory T cells as large as that observed in a normal mouse, and no autoimmune disorder was observed. By contrast, in nonobese diabetic mice, differences in the ability of conventional and regulatory T cells to expand in response to lymphopenia led to an unbalance between these 2 T-cell compartments at the expense of regulatory T cells, resulting in the onset of autoimmune diseases. Notably, this accounted for the rapid transfer of diabetes with small numbers of BDC2.5 CD4(+) T cells. Thus, lymphopenia does not itself induce autoimmunity, but it should be considered as a cofactor for the development of autoimmune disorders. PMID- 19561322 TI - Sex hormones, acting on the TERT gene, increase telomerase activity in human primary hematopoietic cells. AB - Androgens have been used in the treatment of bone marrow failure syndromes without a clear understanding of their mechanism of action. Blood counts of patients with dyskeratosis congenita or aplastic anemia with mutations in telomerase genes can improve with androgen therapy. Here we observed that exposure in vitro of normal peripheral blood lymphocytes and human bone marrow derived CD34(+) cells to androgens increased telomerase activity, coincident with higher TERT mRNA levels. Cells from patients who were heterozygous for telomerase mutations had low baseline telomerase activity, which was restored to normal levels by exposure to androgens. Estradiol had an effect similar to androgens on TERT gene expression and telomerase enzymatic activity. Tamoxifen abolished the effects of both estradiol and androgens on telomerase function, and letrozole, an aromatase inhibitor, blocked androgen effects on telomerase activity. Conversely, flutamide, an androgen receptor antagonist, did not affect androgen stimulation of telomerase. Down-regulation by siRNA of estrogen receptor-alpha (ER alpha), but not ER beta, inhibited estrogen-stimulated telomerase function. Our results provide a mechanism for androgen therapy in bone marrow failure: androgens appear to regulate telomerase expression and activity mainly by aromatization and through ER alpha. These findings have potential implications for the choice of current androgenic compounds and the development of future agents for clinical use. PMID- 19561323 TI - Mobilization in myeloma revisited: IMWG consensus perspectives on stem cell collection following initial therapy with thalidomide-, lenalidomide-, or bortezomib-containing regimens. AB - The past decade has witnessed a paradigm shift in the initial treatment of multiple myeloma with the introduction of novel agents such as thalidomide, lenalidomide, and bortezomib, leading to improved outcomes. High-dose therapy and autologous stem cell transplantation remains an important therapeutic option for patients with multiple myeloma eligible for the procedure. Before the advent of the novel agents, patients underwent stem cell collection prior to significant alkylating agent exposure, given its potential deleterious effect on stem cell collection. With increasing use of the novel agents in the upfront setting, several reports have emerged raising concerns about their impact on the ability to collect stem cells. An expert panel of the International Myeloma Working Group (IMWG) was convened to examine the implications of these therapies on stem collection in patients with myeloma and to develop recommendations for addressing these issues. Here we summarize the currently available data and present our perspective on the problem and potential options to overcome this problem. Specifically, we recommend early mobilization of stem cells, preferably within the first 4 cycles of initial therapy, in patients treated with novel agents and encourage participation in clinical trials evaluating novel approaches to stem cell mobilization. PMID- 19561324 TI - Reintroduction of CEBPA in MN1-overexpressing hematopoietic cells prevents their hyperproliferation and restores myeloid differentiation. AB - Forced expression of MN1 in primitive mouse hematopoietic cells causes acute myeloid leukemia and impairs all-trans retinoic acid-induced granulocytic differentiation. Here, we studied the effects of MN1 on myeloid differentiation and proliferation using primary human CD34(+) hematopoietic cells, lineage depleted mouse bone marrow cells, and bipotential (granulocytic/monocytic) human acute myeloid leukemia cell lines. We show that exogenous MN1 stimulated the growth of CD34(+) cells, which was accompanied by enhanced survival and increased cell cycle traverse in cultures supporting progenitor cell growth. Forced MN1 expression impaired both granulocytic and monocytic differentiation in vitro in primary hematopoietic cells and acute myeloid leukemia cell lines. Endogenous MN1 expression was higher in human CD34(+) cells compared with both primary and in vitro-differentiated monocytes and granulocytes. Microarray and real-time reverse transcribed polymerase chain reaction analysis of MN1-overexpressing CD34(+) cells showed down-regulation of CEBPA and its downstream target genes. Reintroduction of conditional and constitutive CEBPA overcame the effects of MN1 on myeloid differentiation and inhibited MN1-induced proliferation in vitro. These results indicate that down-regulation of CEBPA activity contributes to MN1 modulated proliferation and impaired myeloid differentiation of hematopoietic cells. PMID- 19561325 TI - Rodent and nonrodent malaria parasites differ in their phospholipid metabolic pathways. AB - Malaria, a disease affecting humans and other animals, is caused by a protist of the genus Plasmodium. At the intraerythrocytic stage, the parasite synthesizes a high amount of phospholipids through a bewildering number of pathways. In the human Plasmodium falciparum species, a plant-like pathway that relies on serine decarboxylase and phosphoethanolamine N-methyltransferase activities diverts host serine to provide additional phosphatidylcholine and phosphatidylethanolamine to the parasite. This feature of parasitic dependence toward its host was investigated in other Plasmodium species. In silico analyses led to the identification of phosphoethanolamine N-methyltransferase gene orthologs in primate and bird parasite genomes. However, the gene was not detected in the rodent P. berghei, P. yoelii, and P. chabaudi species. Biochemical experiments with labeled choline, ethanolamine, and serine showed marked differences in biosynthetic pathways when comparing rodent P. berghei and P. vinckei, and human P. falciparum species. Notably, in both rodent parasites, ethanolamine and serine were not significantly incorporated into phosphatidylcholine, indicating the absence of phosphoethanolamine N-methyltransferase activity. To our knowledge, this is the first study to highlight a crucial difference in phospholipid metabolism between Plasmodium species. The findings should facilitate efforts to develop more rational approaches to identify and evaluate new targets for antimalarial therapy. PMID- 19561326 TI - Cerebrospinal immunoglobulin level changes and clinical response to treatment of Hashimoto's encephalopathy. AB - We describe a 64-year-old male who presented with a 2-year history of behavioral and cognitive decline. Brain imaging showed nonenhancing hemispheric white matter lesions. Blood work revealed elevated thyroglobulin and thyroperoxidase antibody levels. Cerebrospinal fluid (CSF) analysis was largely negative, except for an elevated protein and immunoglobulin G (IgG) level. Because of the absence of stroke, central nervous system (CNS) tumor, or infection, this patient fits into criteria of Hashimoto's encephalopathy. His Mini-Mental State Examination score improved from 10 to 29 after initial immunotherapy. The patient remained stable over 6 months with monthly outpatient total plasma exchange, but symptoms recurred within 3 months when the outpatient therapy was discontinued. A follow up CSF IgG level was found to be increased and the treatment was repeated with partial clinical improvement and decline in CSF IgG level. He then underwent high dose steroid treatment after which patients' clinical condition stabilized and CSF analysis showed even further IgG decline. PMID- 19561327 TI - Does exposure to perfluoroalkyl acids present a risk to human health? PMID- 19561328 TI - Commentary: Why diets need to change to avert harm from global warming. PMID- 19561329 TI - Birth cohort studies: past, present and future. PMID- 19561330 TI - Inhibition of mitochondrial fission favours mutant over wild-type mitochondrial DNA. AB - Biased segregation of mitochondrial DNA variants has been widely documented, but little was known about its molecular basis. We set out to test the hypothesis that altering the balance between mitochondrial fusion and fission could influence the segregation of mutant and wild-type mtDNA variants, because it would modify the number of organelles per cell. Therefore human cells heteroplasmic for the pathological A3243G mitochondrial DNA mutation were transfected with constructs designed to silence Drp1 or hFis1, whose gene products are required for mitochondrial fission. Drp1 and hFis1 gene silencing were both associated with increased levels of mutant mitochondrial DNA. Thus, the extent of the mitochondrial reticular network appears to be an important factor in determining mutant load. The fact that the level of mutant and wild-type mitochondrial DNA can be manipulated by altering the expression of nuclear encoded factors involved in mitochondrial fission suggests new interventions for mitochondrial DNA disorders. PMID- 19561331 TI - A novel carotenoid derivative, lutein 3-acetate, accumulates in senescent leaves of rice. AB - The biosynthetic pathway of lutein, the most abundant carotenoid in leaves, has been extensively studied, while its degradation pathway during senescence is poorly understood. We found that a novel carotenoid derivative, lutein 3-acetate, accumulates in senescent leaves of rice (Oryza sativa L.). The change in contents of lutein and lutein 3-acetate suggests that lutein is converted to lutein 3 acetate during senescence. Analysis of mutants involved in light-harvesting complex II (LHCII) accumulation and degradation suggests that the converting activity is induced during senescence and that only free lutein, not lutein bound to LHCII, is converted into lutein 3-acetate. PMID- 19561332 TI - Functional and structural characterization of a flavonoid glucoside 1,6 glucosyltransferase from Catharanthus roseus. AB - Sugar-sugar glycosyltransferases play an important role in structural diversity of small molecule glycosides in higher plants. We isolated a cDNA clone encoding a sugar-sugar glucosyltransferase (CaUGT3) catalyzing 1,6-glucosylation of flavonol and flavone glucosides for the first time from Catharanthus roseus. CaUGT3 exhibited a unique glucosyl chain elongation activity forming not only gentiobioside but also gentiotrioside and gentiotetroside in a sequential manner. We investigated the functional properties of CaUGT3 using homology modeling and site-directed mutagenesis, and identified amino acids positioned in the acceptor binding pocket as crucial for providing enough space to accommodate flavonoid glucosides instead of flavonoid aglycones. These results provide basic information for understanding and engineering the catalytic functions of sugar sugar glycosyltransferases involved in biosynthesis of plant glycosides. PMID- 19561333 TI - Light-dependent and light-independent protochlorophyllide oxidoreductases in the chromatically adapting cyanobacterium Fremyella diplosiphon UTEX 481. AB - The cyanobacterium Fremyella diplosiphon can alternate its light-harvesting pigments, a process called comple-mentary chromatic adaptation (CCA), allowing it to photosynthesize in green light (GL) and in fluctuating light conditions. Nevertheless, F. diplosiphon requires chlorophylls for photosynthesis under all light conditions. Two alternative enzymes catalyze the penultimate step of chlorophyll synthesis, light-dependent protochlorophyllide oxidoreductase (LPOR) and dark-operative protochlo-rophyllide oxidoreductase (DPOR). DPOR enzymatic activity is light independent, while LPOR requires light. Therefore, we hypothesize that F. diplosiphon up-regulates DPOR gene expression in GL, so that DPOR is more abundant when LPOR is less functional. We cloned the genes encoding the three subunits of DPOR, chlL, chlN and chlB, and the LPOR gene, por, to determine the abundance of the transcripts under red light (RL), GL and dark conditions. We found that F. diplosiphon chlL and chlN genes are transcribed as parts of a single operon, a gene structure that is conserved within cyanobacteria. Tran-scripts levels of all DPOR genes are up-regulated approximately 2-fold in GL relative to levels in RL, whereas LPOR transcript levels are reduced in GL. Moreover, mutations in CCA regulators, RcaE and CpeR, modify DPOR and LPOR transcript levels under specific light conditions. Finally, both DPOR and LPOR transcripts are down-regulated 2- to 5-fold in the dark. These results provide the first evidence that light quality and CCA affect the genetic regulation of chlorophyll biosynthesis in freshwater cyanobacteria, ecologically important photosynthetic organisms. PMID- 19561334 TI - Dynamic bridges--a calponin-domain kinesin from rice links actin filaments and microtubules in both cycling and non-cycling cells. AB - Interaction and cross-talk between microtubules and actin microfilaments are important for the cell axis and polarity during plant cell growth and development, but little is known about the molecular components of this interaction. Plant kinesins with a calponin-homology domain (KCHs) were recently identified and associated with a putative role in microtubule-microfilament cross linking. KCHs belong to a distinct branch of the minus end-directed kinesin subfamily and so far have only been identified in land plants including the mosses. Here we report the identification of a new KCH from rice (Oryza sativa), OsKCH1, and show that OsKCH1 is associated with cortical microtubules and actin microfilaments in vivo. Furthermore, OsKCH1 is shown to bind to micro-tubules and actin microfilaments in vitro in a domain-dependent way. Additionally, this unique type of kinesin is shown to oligomerize both in vivo and in vitro. These findings are discussed with respect to a general role for KCHs as linkers between actin filaments and microtubules in both cell elongation and division. PMID- 19561335 TI - ITM Probe: analyzing information flow in protein networks. AB - SUMMARY: Founded upon diffusion with damping, ITM Probe is an application for modeling information flow in protein interaction networks without prior restriction to the sub-network of interest. Given a context consisting of desired origins and destinations of information, ITM Probe returns the set of most relevant proteins with weights and a graphical representation of the corresponding sub-network. With a click, the user may send the resulting protein list for enrichment analysis to facilitate hypothesis formation or confirmation. AVAILABILITY: ITM Probe web service and documentation can be found at www.ncbi.nlm.nih.gov/CBBresearch/qmbp/mn/itm_probe. PMID- 19561336 TI - IMG ER: a system for microbial genome annotation expert review and curation. AB - MOTIVATION: A rapidly increasing number of microbial genomes are sequenced by organizations worldwide and are eventually included into various public genome data resources. The quality of the annotations depends largely on the original dataset providers, with erroneous or incomplete annotations often carried over into the public resources and difficult to correct. RESULTS: We have developed an Expert Review (ER) version of the Integrated Microbial Genomes (IMG) system, with the goal of supporting systematic and efficient revision of microbial genome annotations. IMG ER provides tools for the review and curation of annotations of both new and publicly available microbial genomes within IMG's rich integrated genome framework. New genome datasets are included into IMG ER prior to their public release either with their native annotations or with annotations generated by IMG ER's annotation pipeline. IMG ER tools allow addressing annotation problems detected with IMG's comparative analysis tools, such as genes missed by gene prediction pipelines or genes without an associated function. Over the past year, IMG ER was used for improving the annotations of about 150 microbial genomes. PMID- 19561337 TI - HAPLOWSER: a whole-genome haplotype browser for personal genome and metagenome. AB - SUMMARY: Haplotype assembly is becoming a very important tool in genome sequencing of human and other organisms. Although haplotypes were previously inferred from genome assemblies, there has never been a comparative haplotype browser that depicts a global picture of whole-genome alignments among haplotypes of different organisms. We introduce a whole-genome HAPLotype brOWSER (HAPLOWSER), providing evolutionary perspectives from multiple aligned haplotypes and functional annotations. Haplowser enables the comparison of haplotypes from metagenomes, and associates conserved regions or the bases at the conserved regions with functional annotations and custom tracks. The associations are quantified for further analysis and presented as pie charts. Functional annotations and custom tracks that are projected onto haplotypes are saved as multiple files in FASTA format. Haplowser provides a user-friendly interface, and can display alignments of haplotypes with functional annotations at any resolution. AVAILABILITY: Haplowser, written in Java, supports multiple platforms including Windows and Linux. Haplowser is publicly available at http://embio.yonsei.ac.kr/haplowser . PMID- 19561338 TI - Effects of tolvaptan on dyspnoea relief from the EVEREST trials. AB - AIMS: To describe the effects of tolvaptan therapy on dyspnoea relief based on timing of delivery, influence of concomitant therapies, and baseline patient and clinical characteristics. Also, the influence of clinical trial design on dyspnoea measurement, from the Efficacy of Vasopressin Antagonism in Heart Failure Outcome Study with Tolvaptan (EVEREST) trials. METHODS AND RESULTS: Post hoc analysis was performed based on the endpoint of patient-assessed dyspnoea. Changes from baseline at inpatient Day 1 were compared between treatment groups by the van Elteren test. Pre-determined subgroup analyses were also performed. Tolvaptan's effects are greatest within 12 h after first dose with an additional, but modest dyspnoea improvement benefit irrespective of time after admission. Overall, patients continue to report dyspnoea improvement up to 60 h after admission. The window of enrolment, up to 48 h after admission, combined with measurement on 'Day 1' led to a wide range over when dyspnoea was assessed. CONCLUSION: Post hoc analysis suggests that tolvaptan modestly improves dyspnoea compared with standard therapy alone, regardless if given early or relatively late after hospitalization, and also across major pre-specified subgroups, despite ongoing background therapy aimed at relieving signs and symptoms. Significant variability around when dyspnoea was assessed, in addition to the persistence of dyspnoea despite ongoing background therapy, may influence how future clinical trials assess dyspnoea in acute heart failure syndromes. PMID- 19561339 TI - Cardiomyopathy is associated with structural remodelling of heart valve extracellular matrix. AB - AIMS: To increase the supply, many countries harvest allograft valves from explanted hearts of transplant recipients with ischaemic (ICM) or dilated cardiomyopathy (DCM). This study determines the structural integrity of valves from cardiomyopathic hearts. METHODS AND RESULTS: Extracellular matrix (ECM) was examined in human valves obtained from normal, ICM, and DCM hearts. To confirm if ECM changes were directly related to the cardiomyopathy, we developed a porcine model of chronic ICM. Histology and immunohistostaining, as well as non-invasive multiphoton and second harmonic generation (SHG) imaging revealed marked disruption of ECM structures in human valves from ICM and DCM hearts. The ECM was unaffected in valves from normal and acute ICM pigs, whereas chronic ICM specimens showed ECM alterations similar to those seen in ICM and DCM patients. Proteins and proteinases implicated in ECM remodelling, including Tenascin C, TGFbeta1, Cathepsin B, MMP2, were upregulated in human ICM and DCM, and porcine chronic ICM specimens. CONCLUSION: Valves from cardiomyopathic hearts showed significant ECM deterioration with a disrupted collagen and elastic fibre network. It will be important to determine the impact of this ECM damage on valve durability and calcification in vivo if allografts are to be used from these donors. PMID- 19561340 TI - Three-dimensional imaging of the aortic valve and aortic root with computed tomography: new standards in an era of transcatheter valve repair/implantation. AB - Valvular heart disease is a significant, global cause of morbidity and mortality. In the ageing population of industrialized nations, degenerative aortic stenosis has become the most common native valve disorder. Although surgical aortic valve replacement is associated with excellent clinical outcomes, a large number of patients with severe aortic stenosis are not considered surgical candidates. These patients have poor prognosis with continued medical therapy. In this population, catheter-based approaches for valve replacement/insertion show promising initial results. In an era of expanding catheter-based approaches to valve disease, the requirements for peri-operative imaging are evolving. Because of the lack of direct exposure during the procedure, the operator has to rely increasingly on imaging data rather than direct inspection. Novel three dimensional (3D) imaging modalities, including computed tomography, rapidly acquire volumetric data sets and allow subsequent 3D display and reconstruction in unlimited planes. Procedural planning based on 3D imaging has already become routine for other endovascular procedures including aortic stent grafts, but is in its infancy in the context of transcatheter valve insertion. PMID- 19561341 TI - The unstable plaque: a broken balance. PMID- 19561342 TI - New horizons for in vitro spermatogenesis? An update on novel three-dimensional culture systems as tools for meiotic and post-meiotic differentiation of testicular germ cells. AB - Culture and differentiation of male germ cells has been performed for various purposes in the past. To date, none of the studies aimed at in vitro spermatogenesis has resulted in a sufficient number of mature gametes. Numerous studies have revealed worthy pieces of information, building up a body of information on conditions that are required to maintain and mature male germ cells in vitro. In this review, we report on previously published and unpublished experiments addressing murine germ cell differentiation in three-dimensional (3D) in vitro culture systems. In a systematic set of experiments, we examined the influence of two different matrices (soft agar and methylcellulose) as well as the need for gonadotrophin support. For the first time, we demonstrate that pre meiotic male germ cells [revealed by the absence of meiotic marker expression (e.g. Boule)] obtained from immature mice pass through meiosis in vitro. After several weeks of culture, we obtained morphologically normal spermatozoa embedded in the matrix substance. Complete maturation relied on support from somatic testicular cells and the presence of gonadotrophins but appeared independent from the matrix in a 3D culture environment. Further research efforts are required to reveal the applicability of this culture technique for human germ cells and the functionality of the spermatozoa for generating offspring. PMID- 19561343 TI - The expression of MUC1 in human Fallopian tube during the menstrual cycle and in ectopic pregnancy. AB - BACKGROUND: Ectopic pregnancy is a major cause of maternal morbidity and mortality with increasing incidence worldwide. We have investigated whether epithelia from Fallopian tubes (FTs) bearing an ectopic pregnancy differ from normal tubes in expression of MUC1. METHODS: Since it is not possible to collect FTs from women carrying a healthy pregnancy, we studied tissue collected at the time of hysterectomy for benign disease. Women were injected with hCG in the days leading up to hysterectomy, and pseudopregnancy confirmed by the presence of high serum progesterone levels and the decidualization of the endometrium. FTs from the different stages of the menstrual cycle (n = 24), tubes bearing an ectopic pregnancy (n = 15) and pseudo-pregnant tubes (n = 6) were collected and examined using immunohistochemistry and quantitative RT-PCR. RESULTS: MUC1 was present at the apical surface of the tubal epithelial cells throughout the menstrual cycle, but intracellular localization was minimal in the follicular phase, increasing to a maximum in the luteal phase. MUC1, including the glycoform recognized by antibody 214D4, was found at the apical surface of tubal epithelium in both the ectopic and pseudo-pregnant groups and the intracellular expression was much stronger in the pseudo-pregnant group than in the ectopic group. The 214D4 epitope was absent from tubal tissue adjacent to ectopic implants. CONCLUSIONS: The decrease in MUC1 expression and altered glycosylation in tubal epithelium from ectopic pregnancy may reflect an increase in receptivity. PMID- 19561344 TI - Comparative methylation of ERVWE1/syncytin-1 and other human endogenous retrovirus LTRs in placenta tissues. AB - Human endogenous retroviruses (HERVs) are globally silent in somatic cells. However, some HERVs display high transcription in physiological conditions. In particular, ERVWE1, ERVFRDE1 and ERV3, three proviruses of distinct families, are highly transcribed in placenta and produce envelope proteins associated with placenta development. As silencing of repeated elements is thought to occur mainly by DNA methylation, we compared the methylation of ERVWE1 and related HERVs to appreciate whether HERV methylation relies upon the family, the integration site, the tissue, the long terminal repeat (LTR) function or the associated gene function. CpG methylation of HERV-W LTRs in placenta-associated tissues was heterogeneous but a joint epigenetic control was found for ERVWE1 5'LTR and its juxtaposed enhancer, a mammalian apparent LTR retrotransposon. Additionally, ERVWE1, ERVFRDE1 and ERV3 5'LTRs were all essentially hypomethylated in cytotrophoblasts during pregnancy, but showed distinct and stage-dependent methylation profiles. In non-cytotrophoblastic cells, they also exhibited different methylation profiles, compatible with their respective transcriptional activities. Comparative analyses of transcriptional activity and LTR methylation in cell lines further sustained a role for methylation in the control of functional LTRs. These results suggest that HERV methylation might not be family related but copy-specific, and related to the LTR function and the tissue. In particular, ERVWE1 and ERV3 could be developmentally epigenetically regulated HERVs. PMID- 19561345 TI - A randomized prospective study comparing two flexible epidural catheters for labour analgesia. AB - BACKGROUND: Previous studies evaluating stiff epidural catheters found that the three-holed design provided superior labour analgesia compared with an end-holed design. This was believed due to improved medication distribution. Recently, flexible epidural catheters with both designs have been shown to be superior to the stiff epidural catheters. We investigated the success of labour analgesia comparing the flexible three-holed with the flexible end-holed epidural catheter. METHODS: This was a prospective, single-blinded randomized study. We enrolled 500 parturients in active labour. The primary outcome was complete relief of labour pain assessed at 30 min. We also assessed the occurrence of paresthesias, intravascular and intrathecal placement, catheter replacement, and treatment of breakthrough pain during labour. Comparisons were made using Pearson's chi(2), with significance determined at the 0.05 level. RESULTS: Four hundred and ninety three subjects completed the study. Initial analgesia was similar (complete labour analgesia: end-holed=85% vs 80% 95% CI of difference: 13% to -3%; P=NS). The incidence of paresthesia was similar (end-holed=3.6% vs 5.3%; P=NS). There was one intrathecal and three intravascular catheters in the three-holed group and two intravascular catheters in the end-holed group. The number of supplemental boluses and catheter replacements required during labour was similar between the groups. CONCLUSIONS: There were no differences in the initial analgesia success rate, complications, or labour analgesia between end-hole vs multi-hole flexible epidural catheters. PMID- 19561346 TI - Developmental changes in spatial distribution of in vivo fluorescence and epidermal UV absorbance over Quercus petraea leaves. AB - BACKGROUND AND AIMS: Epidermal phenolic compounds (mainly flavonoids) constitute a vital screen that protects the leaf from damage by natural ultraviolet (UV) radiation. The effectiveness of epidermal UV-screening depends on leaf anatomy, the content of UV-screening compounds and their spatial uniformity over the leaf area. To investigate in vivo the spatial pattern of the epidermal UV-screen during leaf development, a fluorescence imaging method was developed to map the epidermal UV-absorbance at a microscopic scale. This study was done on oak (Quercus petraea) leaves that were used as a model of woody dicotyledonous leaves. METHODS: The leaf development of 2-year-old trees, grown outdoors, was monitored, at a macroscopic scale, by in vivo measurements of chlorophyll content per unit area and epidermal UV-absorbance using two optical leaf-clip meters. The distribution of pigments within leaves was assessed in vivo spectroscopically. The microscopic images of UV-induced fluorescence and UV-absorbance acquired in vivo during leaf development were interpreted from spectral characteristics of leaves. KEY RESULTS: At a macroscopic scale, epidermal UV-absorbance was high on the upper leaf side during leaf development, while it increased on the lower leaf side during leaf expansion and reached the adaxial value at maturity. At a microscopic scale, in immature leaves, for both leaf sides, the spatial distribution of epidermal UV-absorbance was heterogeneous, with a pattern depending on the flavonoid content of vacuoles in developing epidermal cells. At maturity, epidermal UV-absorbance was uniform. CONCLUSIONS: The spatial pattern of epidermal UV-screen over the area of oak leaves is related to leaf anatomy during development. In vivo spectroscopy and fluorescence imaging of the leaf surface showed the distribution of pigments within the leaf and hence can provide a tool to monitor optically the leaf development in nature. PMID- 19561347 TI - Progress with the European Society for Paediatric Nephrology (ESPN)/ERA-EDTA Registry for children with established renal failure (ERF). PMID- 19561349 TI - Induction of anaesthesia with either midazolam or propofol in rabbits premedicated with fentanyl/fluanisone. AB - Fifty-four rabbits that were to be neutered were premedicated with 0.1 ml/kg fentanyl/fluanisone and then randomly allocated to be anaesthetised with either midazolam or propofol. Anaesthesia was then maintained with isoflurane. The ease of orotracheal intubation, the rabbits' cardiorespiratory variables, and the speed and quality of recovery from anaesthesia were assessed by the same anaesthetist who was unaware of the induction agent used. Hypotension was common in both groups. The mean (sd) respiratory rates were 30 (12) breaths per minute in the midazolam group and 43 (15) breaths per minute in the propofol group. The mean (sd) time to first head lift was 36 (21) minutes in the midazolam group and 14 (11) minutes in the propofol group, and the mean (sd) times to the return of the righting reflex were 35 (19) minutes and 15 (eight) minutes, respectively. The quality of recovery was better in the propofol group than in the midazolam group. PMID- 19561350 TI - Beneficial effects on the reproductive performance of sows of administering prostaglandin analogues after farrowing. AB - Twenty-four to 36 hours after farrowing, 192 sows were treated with a single intramuscular injection (2 ml per animal) of a prostaglandin analogue; 102 were treated with cloprostenol racemate and 97 with dinoprost tromethamine, and 90 were left untreated. There were no statistically significant differences between the groups in the percentages of sows that came into oestrus by eight days after weaning or conceived by eight days after weaning. Significantly more piglets were born per litter (10.71 and 11.00 piglets in the cloprostenol and dinoprost groups, respectively) and born alive (10.22 and 10.41, respectively) than in the controls (9.24 piglets born per litter and 8.66 piglets born alive). PMID- 19561351 TI - Scintigraphic pattern of uptake of 99mTechnetium by the cervical vertebrae of sound horses. AB - Lateral scintigraphic images of the necks of 12 show jumping horses, 12 thoroughbreds and 12 French trotters were evaluated. Custom-written automatic analysis software was used to segment the regions of the articular processes and vertebral symphyses from C3 to C7 and to calculate their mean radiopharmaceutical uptakes. The uptakes by the two sides of these sound horses were not significantly different, and the uptakes by the vertebral symphyses and articular processes were also similar in the three groups of horses, considering each vertebral level separately. In the thoroughbred race horses, the mean uptake by the C6/C7 symphyses was significantly higher than by C5/C6, and the mean uptake by the C6/C7 articular processes was significantly higher than by C3/C4 and C4/C5. These differences were not observed in the French trotters and show jumping horses, but they were observed when the data for the three groups of horses were combined. PMID- 19561352 TI - Prevalence of equine gastric ulceration syndrome in standardbreds. PMID- 19561353 TI - Clostridium perfringens type D epsilon intoxication in one-day-old calves. PMID- 19561354 TI - Use of antimicrobials. PMID- 19561355 TI - Proposed changes to veterinary legislation. PMID- 19561356 TI - Predator attacks on sheep in Northumberland. PMID- 19561357 TI - Deglutathionylation of 2-Cys peroxiredoxin is specifically catalyzed by sulfiredoxin. AB - Reversible protein glutathionylation plays a key role in cellular regulation and cell signaling and protects protein thiols from hyperoxidation. Sulfiredoxin (Srx), an enzyme that catalyzes the reduction of Cys-sulfinic acid derivatives of 2-Cys peroxiredoxins (2-Cys Prxs), has been shown to catalyze the deglutathionylation of actin. We show that deglutathionylation of 2-Cys Prx, a family of peroxidases, is specifically catalyzed by Srx. Using the ubiquitously expressed member of 2-Cys Prx, Prx I, we revealed the following. (i) Among its four Cys residues, Cys(52), Cys(83), and Cys(173) can be glutathionylated in vitro. Deglutathionylation with Cys mutants showed that Cys(83) and Cys(173) were preferentially catalyzed by Srx, with glutathionylated Srx as the reaction intermediate, whereas glutaredoxin I was more favorable for deglutathionylating Cys(52). (ii) Studies using site-directed mutagenesis coupled with binding and deglutathionylation activities revealed that Pro(174) and Pro(179) of Prx I and Tyr(92) of Srx are essential for both activities. Furthermore, relative to glutaredoxin I, Srx exhibited negligible deglutathionylation activity for glutathionylated cysteine and glutathionylated BSA. These results indicate that Srx is specific for deglutathionylating Prx I due to its favorable affinity for Prx I. To assess the biological relevance of these observations, we showed that Prx I is glutathionylated in A549 and HeLa cells under modest levels of H(2)O(2). In addition, the level of glutathionylated Prx I was substantially elevated in small interfering RNA-mediated Srx-knocked down cells, whereas the reverse was observed in Srx-overexpressing cells. However, glutathionylation of Prx V, not known to bind to Srx, was not affected by the change in Srx expression levels. PMID- 19561358 TI - FANCI protein binds to DNA and interacts with FANCD2 to recognize branched structures. AB - In this study, we report that the purified wild-type FANCI (Fanconi anemia complementation group I) protein directly binds to a variety of DNA substrates. The DNA binding domain roughly encompasses residues 200-1000, as suggested by the truncation study. When co-expressed in insect cells, a small fraction of FANCI forms a stable complex with FANCD2 (Fanconi anemia complementation group D2). Intriguingly, the purified FANCI-FANCD2 complex preferentially binds to the branched DNA structures when compared with either FANCI or FANCD2 alone. Co immunoprecipitation with purified proteins indicates that FANCI interacts with FANCD2 through its C-terminal amino acid 1001-1328 fragment. Although the C terminus of FANCI is dispensable for direct DNA binding, it seems to be involved in the regulation of DNA binding activity. This notion is further enhanced by two C-terminal point mutations, R1285Q and D1301A, which showed differentiated DNA binding activity. We also demonstrate that FANCI forms discrete nuclear foci in HeLa cells in the absence or presence of exogenous DNA damage. The FANCI foci are colocalized perfectly with FANCD2 and partially with proliferating cell nuclear antigen irrespective of mitomycin C treatment. An increased number of FANCI foci form and become resistant to Triton X extraction in response to mitomycin C treatment. Our data suggest that the FANCI-FANCD2 complex may participate in repair of damaged replication forks through its preferential recognition of branched structures. PMID- 19561359 TI - The interaction of mitochondrial iron with manganese superoxide dismutase. AB - Superoxide dismutase 2 (SOD2) is one of the rare mitochondrial enzymes evolved to use manganese as a cofactor over the more abundant element iron. Although mitochondrial iron does not normally bind SOD2, iron will misincorporate into Saccharomyces cerevisiae Sod2p when cells are starved for manganese or when mitochondrial iron homeostasis is disrupted by mutations in yeast grx5, ssq1, and mtm1. We report here that such changes in mitochondrial manganese and iron similarly affect cofactor selection in a heterologously expressed Escherichia coli Mn-SOD, but not a highly homologous Fe-SOD. By x-ray absorption near edge structure and extended x-ray absorption fine structure analyses of isolated mitochondria, we find that misincorporation of iron into yeast Sod2p does not correlate with significant changes in the average oxidation state or coordination chemistry of bulk mitochondrial iron. Instead, small changes in mitochondrial iron are likely to promote iron-SOD2 interactions. Iron binds Sod2p in yeast mutants blocking late stages of iron-sulfur cluster biogenesis (grx5, ssq1, and atm1), but not in mutants defective in the upstream Isu proteins that serve as scaffolds for iron-sulfur biosynthesis. In fact, we observed a requirement for the Isu proteins in iron inactivation of yeast Sod2p. Sod2p activity was restored in mtm1 and grx5 mutants by depleting cells of Isu proteins or using a dominant negative Isu1p predicted to stabilize iron binding to Isu1p. In all cases where disruptions in iron homeostasis inactivated Sod2p, we observed an increase in mitochondrial Isu proteins. These studies indicate that the Isu proteins and the iron-sulfur pathway can donate iron to Sod2p. PMID- 19561360 TI - Docosahexaenoic acid reduces suppressive and migratory functions of CD4+CD25+ regulatory T-cells. AB - Immunological tolerance is one of the fundamental aspects of the immune system. The CD4(+)CD25(+) regulatory T (Treg) cells have emerged as key players in the development of tolerance to self and foreign antigens. However, little is known about the endogenous factors and mechanisms controlling their suppressive capacity on immune response. In this study, we observed that docosahexaenoic acid (DHA), an n-3 polyunsaturated fatty acid, diminished, in a dose-dependent manner, the capacity of Treg cells to inhibit the CD4(+)CD25(-) effector T-cell proliferation. DHA not only reduced the migration of Treg cells toward chemokines but also downregulated the mRNA expression of CCR-4 and CXCR-4 in Treg cells. DHA also curtailed ERK1/2 and Akt phosphorylation and downregulated the Smad7 levels in these cells. Contradictorily, DHA upregulated the mRNA expression of Foxp3, CTLA-4, TGF-beta, and IL-10; nonetheless, this fatty acid increased the expression of p27(KIP1) mRNA, known to be involved in Treg cell unresponsiveness. In Foxp3-immunoprepitated nuclear proteins, DHA upregulated histone desacetylase 7 levels that would again participate in the unresposnsiveness of these cells. Finally, a DHA-enriched diet also diminished, ex vivo, the suppressive capacity of Treg cells. Altogether, these results suggest that DHA, by diminishing Treg cell functions, may play a key role in health and disease. PMID- 19561361 TI - EULAR Sjogren's syndrome disease activity index: development of a consensus systemic disease activity index for primary Sjogren's syndrome. AB - OBJECTIVE: To develop a disease activity index for patients with primary Sjogren's syndrome (SS): the European League Against Rheumatism (EULAR) Sjogren's syndrome disease activity index (ESSDAI). METHODS: Thirty-nine SS experts participated in an international collaboration, promoted by EULAR, to develop the ESSDAI. Experts identified 12 organ-specific 'domains' contributing to disease activity. For each domain, features of disease activity were classified in three or four levels according to their severity. Data abstracted from 96 patients with systemic complications of primary SS were used to generate 702 realistic vignettes for which all possible systemic complications were represented. Using the 0-10 physician global assessment (PhGA) scale, each expert scored the disease activity of five patient profiles and 20 realistic vignettes. Multiple regression modelling, with PhGA used as the dependent variable, was used to estimate the weight of each domain. RESULTS: All 12 domains were significantly associated with disease activity in the multivariate model, domain weights ranged from 1 to 6. The ESSDAI scores varied from 2 to 47 and were significantly correlated with PhGA for both real patient profiles and realistic vignettes (r=0.61 and r=0.58, respectively, p<0.001). Compared with 57 (59.4%) of the real patient profiles, 468 (66.7%) of the realistic vignettes were considered likely or very likely to be true. CONCLUSIONS: The ESSDAI is a clinical index designed to measure disease activity in patients with primary SS. Once validated, such a standardised evaluation of primary SS should facilitate clinical research and be helpful as an outcome measure in clinical trials. PMID- 19561362 TI - Impact of graphic and text warnings on cigarette packs: findings from four countries over five years. AB - OBJECTIVES: To examine the impact of health warnings on smokers by comparing the short-term impact of new graphic (2006) Australian warnings with: (i) earlier (2003) United Kingdom larger text-based warnings; (ii) and Canadian graphic warnings (late 2000); and also to extend our understanding of warning wear-out. METHODS: The International Tobacco Control Policy Evaluation Survey (ITC Project) follows prospective cohorts (with replenishment) of adult smokers annually (five waves: 2002-2006), in Canada, United States, UK and Australia (around 2000 per country per wave; total n = 17 773). Measures were of pack warning salience (reading and noticing); cognitive responses (thoughts of harm and quitting); and two behavioural responses: forgoing cigarettes and avoiding the warnings. RESULTS: All four indicators of impact increased markedly among Australian smokers following the introduction of graphic warnings. Controlling for date of introduction, they stimulated more cognitive responses than the UK (text-only) changes, and were avoided more, did not significantly increase forgoing cigarettes, but were read and noticed less. The findings also extend previous work showing partial wear-out of both graphic and text-only warnings, but the Canadian warnings have more sustained effects than UK ones. CONCLUSIONS: Australia's new health warnings increased reactions that are prospectively predictive of cessation activity. Warning size increases warning effectiveness and graphic warnings may be superior to text-based warnings. While there is partial wear-out in the initial impact associated with all warnings, stronger warnings tend to sustain their effects for longer. These findings support arguments for governments to exceed minimum FCTC requirements on warnings. PMID- 19561363 TI - Preoperative 6-minute walk test adds prognostic information to Euroscore in patients undergoing aortic valve replacement. AB - AIMS: The authors investigated the additive prognostic value of the 6-minute walk test (6MWT) to Euroscore in patients with severe aortic stenosis undergoing aortic valve replacement (AVR) METHODS AND RESULTS: 208 patients with severe AS underwent the 6MWT before AVR, as part of a randomised trial (ASSERT) comparing stented and stentless aortic valves. Clinical follow-up was available for 200 patients up to 12 months. The rate of death, myocardial infarction (MI) or stroke (time to first event) was 13% (n = 14) in patients walking <300 metres compared to 4% (n = 4) in those who walked > or =300 metres (p = 0.017). When rate of death, MI or stroke by Euroscore risk was stratified by 6-minute walking distance, the 6MWT added prognostic information. In a Cox regression analysis 6MWT distance was the only variable retained as an independent predictor of the composite outcome of death, MI or stroke at 12 months (HR 0.28 95% CI 0.09 to 0.85, p = 0.025). CONCLUSIONS: The 6MWT is safe and feasible to carry out in patients with severe aortic stenosis before AVR, and provides potentially important functional and prognostic information to clinical assessment and the Euroscore risk score. PMID- 19561364 TI - Anthropometric assessment of abdominal obesity and coronary heart disease risk in men: the PRIME study. AB - OBJECTIVE: Waist-to-height ratio is an anthropometric indicator of abdominal obesity that accounts for stature. Earlier studies have reported marked associations between the waist-to-height ratio and cardiovascular risk factors. The goal of this study was to compare the associations of waist-to-height ratio, waist girth, waist-to-hip ratio or body mass index (BMI) with incidence of coronary events. DESIGN: Prospective study with 10 602 men, aged 50-59 years, recruited between 1991 and 1993 in three centres in France and one centre in Northern Ireland. Clinical and biological data were obtained at interview by trained staff. During the 10 years of follow-up 659 incident coronary events (CHD) were recorded. The relations between anthropometric markers and coronary events were estimated by Cox proportional hazards models. RESULTS: Waist circumference, waist-to-hip ratio, waist-to-height ratios and BMI were positively associated with blood pressure (p<0.0001), diabetes (p<0.0001), low-density lipoprotein (LDL)-cholesterol (p<0.0001), triglycerides (p<0.0001) and inversely correlated to high-density lipoprotein (HDL)-cholesterol (p<0.0001). There was a linear association between waist circumference, waist-to-hip ratio, waist-to height ratio, BMI and CHD events. The age-adjusted and centre-adjusted relative risks (95% CI) for CHD were 1.57 (1.22 to 2.01), 1.75 (1.34 to 2.87), 2.3 (1.79 to 2.99) and 1.99 (1.54 to 2.56) in the 5th quintile vs the first quintile of waist circumference, waist-to-hip ratio, waist-to-height ratio and BMI distribution, respectively. After further adjustment for school duration, physical activity, tobacco and alcohol consumption, hypertension, diabetes, HDL cholesterol and triglycerides, the relative risks for CHD were 0.99 (0.76 to 1.30) for waist circumference (p = 0.5), 1.22 (0.93 to 1.60) for waist-to-hip ratio (p = 0.1), 1.53 (1.16 to 2.01) for waist-to-height ratio (p = 0.03) and 1.30 (0.99 to 1.71) for BMI (p = 0.06). CONCLUSION: In middle-aged European men, waist-to-height ratio identifies coronary risk more strongly than waist circumference, waist-to-hip ratio or BMI, though the difference is marginal. PMID- 19561365 TI - Acute-exercise-induced alterations in calpain and calpastatin expression in rat muscle. AB - CONTEXT: Calpains and calpastatin can degrade muscle proteins, but no research has investigated the expression pattern of calpains and calpastatin after exhaustive exercise. OBJECTIVE: To investigate the alterations in expression of micro-, m-, and n-calpain and calpastatin after exhaustive exercise and its association with muscle injury. METHOD: 64 rats divided into 2 groups, a nonexercise control group and an acute-exhaustive-exercise (AEE) group. Biopsies in the AEE group were taken at different times after exercise. RESULTS: Calpastatin protein expression and m-calpain activity increased early after exercise, but both n-calpain protein expression and micro-calpain activity generally decreased with time. n-Calpain mRNA expression was down-regulated from late after exercise. CONCLUSIONS: The increased m-calpain activity might promote muscle-protein degradation and muscle injury. On the contrary, calpastatin might execute a protective function against muscle injury. The change in micro-calpain activity was found earlier than muscle injury and therefore might serve as a useful predictor of muscle injury. PMID- 19561366 TI - The effect of scapular-retractor fatigue on external and internal rotation in patients with internal impingement. AB - CONTEXT: Scapular strengthening is thought to be an important component of the rehabilitation of patients with internal impingement. OBJECTIVE: To determine the effect of scapular-retractor-muscle fatigue on internal- and external-rotation torque production in patients with internal impingement. DESIGN: Case control study. SETTING: Outpatient clinic. PARTICIPANTS: 15 patients and 18 healthy subjects. INTERVENTION: A scapular-retractor-fatigue protocol. MAIN OUTCOME MEASURE: Shoulder-rotation-torque production. RESULTS: After the scapular retractor-fatigue protocol external-rotation strength was reduced in patients (involved 25%, noninvolved 19%; P < .001). CONCLUSION: Fatigue in the scapular retractors resulted in lower shoulder-rotation-torque production. These findings emphasize the importance of the scapular retractors for proper function of the shoulder rotators with the arm in an abducted position in patients with internal impingement. PMID- 19561367 TI - The effects of gender and fatigue on dynamic postural control. AB - CONTEXT: Deficits in static postural control related to fatigue have been investigated previously, but there is little evidence to link fatigue to performance measures of dynamic postural control. OBJECTIVE: To investigate the effects of fatigue and gender on performance measures of the Star Excursion Balance Test (SEBT). DESIGN: Mixed-model design. SETTING: Research laboratory. PARTICIPANTS: 16 healthy young adults. INTERVENTION: Subjects performed the SEBT before and after 4 different fatiguing conditions. MAIN OUTCOME MEASURES: The normalized reach distances and sagittal-plane kinematics of the knee and hip were recorded. RESULTS: Fatigue produced deficits in normalized reach distances and decreased knee flexion in all 3 reaching directions. Overall, women were able to reach farther than men while simultaneously demonstrating a greater amount of knee flexion. CONCLUSIONS: Gender differences were observed during performance of the SEBT, with women demonstrating greater reach distances and knee flexion, and fatigue amplified these differences. PMID- 19561368 TI - Psychological benefits of virtual reality for patients in rehabilitation therapy. AB - CONTEXT: Whether virtual rehabilitation is beneficial has not been determined. OBJECTIVE: To investigate the psychological benefits of virtual reality in rehabilitation. DESIGN: An experimental group underwent therapy with a virtual reality-based exercise bike, and a control group underwent the therapy without virtual-reality equipment. SETTING: Hospital laboratory. PATIENTS: 30 patients suffering from spinal-cord injury. INTERVENTION: A designed rehabilitation therapy. MAIN OUTCOME MEASURES: Endurance, Borg's rating-of-perceived-exertion scale, the Activation-Deactivation Adjective Check List (AD-ACL), and the Simulator Sickness Questionnaire. RESULTS: The differences between the experimental and control groups were significant for AD-ACL calmness and tension. CONCLUSION: A virtual-reality-based rehabilitation program can ease patients' tension and induce calm. PMID- 19561369 TI - Laboratory gait analysis in patients with low back pain before and after a pilates intervention. AB - OBJECTIVE: To evaluate the influence of pain on vertical ground-reaction force (VGRF) in patients with low back problems and the effect of the Pilates method on the gait of these patients. DESIGN: A single-blind randomized controlled trial. PARTICIPANTS: 28 individuals assigned to a control group (n = 11) and a low-back group (n = 17), the latter of which was subdivided into a Pilates group (n = 8) and a no-Pilates group (n = 9). INTERVENTION: The Pilates group undertook 15 sessions of Pilates. MAIN OUTCOME MEASURES: The VGRF parameters were recorded during preferred and faster walking speeds. The data were collected before and after the intervention. RESULTS: The weight-acceptance rate and push-off rate were significantly less in the right lower limb of low-back group than of the control group at preferred speed. Improvements were seen in the Pilates group postintervention, with increased middle-support force for the left lower limb at faster walking speed and decreased pain; this did not occur in the no-Pilates group. CONCLUSIONS: These results suggest that patients with low back pain use strategies to attenuate the amount of force imposed on their body. The Pilates method can improve weight discharge in gait and reduce pain compared with no intervention. PMID- 19561370 TI - Temporal values and plantar pressures during normal walking and racewalking in a group of racewalkers. AB - OBJECTIVES: To compare temporal and pressure values between racewalking and normal walking (freely selected speed) and evaluate the impact of racewalking on normal walking. DESIGN AND PARTICIPANTS: Temporal and plantar-pressure values were recorded (xPression system) during normal walking and racewalking in 8 high level racewalkers. The Wilcoxon test was used for comparisons. MEASUREMENTS: Duration of walking and racewalking cycle phases (seconds and percentage of the cycle), peak and average pressures under the hind foot and metatarsal heads, and pressure distribution (%) among metatarsal heads. RESULTS AND CONCLUSIONS: Normal walking: temporal parameters similar to those described in normal gait; peak pressures higher than those described in nonracewalkers with displacement toward lateral forefoot. Racewalking: shorter cycles (important decrease of midstance phase); higher peak pressures than during normal walking in the hind foot and 4th and 5th metatarsal heads; average pressures similar to normal walking in hind foot and lower in forefoot; pressure displacement toward lateral forefoot greater than in normal walking. PMID- 19561371 TI - Reliability of a digital-photographic-goniometric method for coronal-plane lower limb measurements. AB - CONTEXT: Structural and coronal-plane-alignment characteristics of the lower limb are frequently cited as factors contributing to knee pathologies. OBJECTIVE: The purpose of this study was to determine the accuracy and reliability characteristics of a digital-photographic-goniometric method (DPGM) of measurement for 2-dimensional (2D) coronal-plane lower limb measurements of the quadriceps (Q) angle, tibiofemoral (TF) angle, and femur length in human participants adopting a self-selected- or Romberg-stance position. DESIGN: Reliability study. SETTING: University motion-analysis laboratory. PARTICIPANTS: A convenience sample of 20 healthy young adult men and women. MAIN OUTCOME MEASURES: Intraclass correlation coefficients (ICCs), 95% confidence intervals, and standard error of the measurements. RESULTS: Intratester- and intertester reliability coefficients for the Q angle (ICCs .458-.845 and .257-.737) were consistently lower than those for the TF angle (ICCs .627-.904 and .700-.839) or femur length (ICCs .867-.958 and .866-.944). Q angles were also significantly larger (13.4%) in the Romberg- vs self-selected-stance position (P < .001) and larger (20.2%) in the left limb than the right limb. CONCLUSIONS: The DPGM has the potential to produce accurate and reliable measurements of selected 2D lower limb measures. However, the reliability characteristics depend on the ability of the testers to correctly and repeatably landmark the anatomical sites used to define the measurements of interest and might be influenced by other factors such as the stance position adopted, the complexity of the variable (ie, number of anatomical landmarks and segments), and the size of the captured image. Further investigation of these latter factors is warranted. PMID- 19561372 TI - Comparison of training between 2 cycle ergometers on dynamic balance for middle aged women. AB - CONTEXT: Interval cycle training could positively influence dynamic balance in middle-aged women. OBJECTIVE: To compare training effects of a strength ergometer and a standard ergometer on 3 dynamic balance tests. DESIGN: Repeated measures. SETTING: Laboratory. PARTICIPANTS: Seventeen women were randomly assigned to standard (n = 10) or strength cycle ergometry (n = 7). A control group consisted of 7 women. INTERVENTION: Ergometry interval training (3 sessions/wk for 4 wk). MAIN OUTCOME MEASURES: Three balance tests-the Star Excursion Balance Test (SEBT), timed up-and-go (TUG), and four-square step test (FSST)-were performed at pretraining and 4 wk posttraining. RESULTS: Four SEBT directions improved and faster scores for FSST and TUG tests for the standard-cycle group were found, whereas the strength-cycle group only improved their TUG scores. No changes posttraining for the control group. CONCLUSIONS: Stationary cycle training should be included in the dynamic balance-rehabilitation protocol for middle-aged women. PMID- 19561373 TI - How back-muscle fatigue influences lumbar curvature. AB - CONTEXT: Back-muscle fatigue and lumbar curvature are related to low back pain, but the relation between them is unknown. OBJECTIVE: To clarify whether lumbar curvature changes with back-muscle fatigue. DESIGN: Prospective pseudorandomized. SETTING: Laboratory. PARTICIPANTS: 13 healthy men without low back pain. INTERVENTION: To induce back-muscle fatigue, subjects performed a repetitive prone back-extension exercise from 45 degrees to 0 degrees until exhaustion and then rested 10 min. They performed this exercise 3 times. MAIN OUTCOME MEASURES: Before the trial and after each exercise, lumbar curvature was measured with a Spinal Mouse. RESULTS: After back-muscle fatigue, lumbar curvature decreased significantly (P < .01). This decrease occurred at L4-S1, which was defined as the lower spine. On the contrary, curvature of the upper lumbar spine, at Th12 L4, did not decrease. CONCLUSION: Lumbar curvature, especially the lower portion, decreased with back-muscle fatigue induced by prone back-extension exercise. PMID- 19561374 TI - The effect of beta interferon on dendritic cells and cytokine synthesis by CD4+ T cells. AB - BACKGROUND: Dendritic cells (DC) are a key regulator of the immune response, and interferon-beta (IFN-beta) is considered an immunomodulatory molecule for DC. OBJECTIVE: The purpose of this study was to evaluate the ability of IFN-beta treated DC to induce cytokine secretion by CD4+ T cells. METHODS: Dendritic cells were generated from blood monocytes with granulocyte-monocyte colony-stimulating factor and interleukin-4 with or without IFN-beta. We analyzed the production of CD4+ T helper cytokines (IL-17, IFN-gamma and IL-10) in the supernatant of the dendritic cell-T cell co- cultures by ELISA. We also studied the effects of HLA-G and costimulatory molecules on immature and mature DC. RESULTS: IFN-gamma and IL 17 decreased significantly in the presence of HLA-G-bearing DC compared to control cultures (p<0.05). CONCLUSION: Using the mixed leukocyte reaction, we found that DC treated with IFN-beta mediated the inhibition of T cell activation via cytokine production. We conclude that this is important for preventing overactivation of the immune system. PMID- 19561375 TI - The effects of Candida albicans cell wall protein fraction on dendritic cell maturation. AB - BACKGROUND: Candida albicans is a member of the normal human microflora. C. albicans cell wall is composed of several protein and carbohydrate components which have been shown to play a crucial role in C. albicans interaction with the host immune system. Major components of C. albican cell wall are carbohydrates such as mannans, beta glucans and chitins, and proteins that partially modulate the host immune responses. Dendritic cells (DC), as the most important antigen presenting cells of the immune system, play a critical role in inducing immune responses against different pathogens. OBJECTIVE: We investigated the effect of the cell wall protein fraction (CPF) of C. albicans on DC maturation. METHODS: The CPF of C. albicans cells was extracted by a lysis buffer containing sodium dodecyl sulphate, 2-mercaptoethanol and phosphate-buffered saline. The extract was dialyzed and its protein pattern was evaluated by electrophoresis. Dendritic cells were purified from Balb/c mice spleens through a three-step method including mononuclear cell separation, as well as 2-h and overnight cultures. The purified CPF was added at different concentrations to DC. The purity and maturation status of DC were determined by flow cytometry using monoclonal antibodies against CD11c, MHC-II, CD40 and CD86. RESULTS: Treatment of DC with 10 microg/ml of CPF increased the expression of maturation markers including MHC-II, CD86 and CD40 on DC compared to the control group. CONCLUSION: In this study we used C. albicans CPF with the molecular weight of 40-45 kDa for pulsing and maturation of dendritic cells. Since according to our results CPF significantly increased the expression of maturation markers on DC, we suggest that CPF may act as an efficient immunomodulator, or may be used as a potential adjuvant to boost the host immune system against infections. PMID- 19561376 TI - Expression of recombinant heat-shock protein 70 of MCAN/IR/96/LON-49, a tool for diagnosis and future vaccine research. AB - BACKGROUND: Heat shock protein 70 (HSP70) is present in all organisms studied so far, and is a major immunogen in infections caused by pathogens including Leishmania spp. OBJECTIVE: The aim of this study was to clone and express HSP70 from L. infantum strain MCAN/IR/96/LON-49 and evaluate antibody response against HSP70 in visceral leishmaniasis (VL). METHODS: The L. infantum HSP70 gene segment was amplified by specific primers. It was cloned into pTZ57R vector and subcloned into pET32a (+) expression vector. The new construct was transformed in the E.coli Rosetta strain, and HSP70 protein was expressed in the presence of 1 mM IPTG and purified using a HiTrap chelating column. Antibody responses against HSP70 were determined by ELISA in 37 patients with visceral leishmaniasis and 63 healthy controls. RESULTS: Expression of HSP70 protein was confirmed using SDS PAGE electrophoresis and dot blot with an anti-His tag antibody. There was no difference between the sequence of nucleotides of the HSP70 gene in the present study and other reported sequences. The ELISA results indicated that the sera of 81.1% (30/37) of the patients and 6.3% (5/63) of controls reacted to L. infantum HSP70. CONCLUSION: The conservative nature of the HSP70 molecule is an advantage in vaccine studies, because of minor differences (6%) between the nucleotide sequences and consequently the similarity in amino acid sequences in various strains of L. infantum. It could therefore be used in vaccine research against leishmaniasis and also as a tool for serodiagnosis. PMID- 19561377 TI - Ganoderma lucidum induces the expression of CD40/CD86 on peripheral blood monocytes. AB - BACKGROUND: The major immuno-modulating effects of Ganoderma lucidum include mitogenicity and activation of immune effector cells such as T cells, macrophages and natural killer cells resulting in the production of cytokines. OBJECTIVE: The purpose of this study was to evaluate the expression of CD40 and CD80 by G. lucidum-treated human peripheral blood mononuclear cells. METHODS: Monocytes were isolated and incubated at 37 C and 5% CO2 for 24 h and 48 h in the presence or absence of different concentrations of G. lucidum. Cells were then incubated with labelled monoclonal antibodies against CD14, CD40 and B7-1(CD80) molecules utilizing standard protocols, and analyzed by flow cytometry. RESULTS: The results showed that incubation of monocytes with G. lucidum led to marked enhancement of CD40 and B7-1 expression in a dose- and time- dependent manner (p<0.001). G. lucidum was more effective in enhancing the expression of CD80 and CD40 molecules of cells obtained from females than male donors (p<0.001). CONCLUSION: G. lucidum enhanced the expression of CD40 and CD80 molecules on peripheral blood monocytic cells derived from both sexes in a dose-dependent manner, with a preferential higher effect on cells obtained from female donors. PMID- 19561378 TI - Evaluation of plasma interleukin-8 concentration in patients with prostate cancer and benign prostate hyperplasia. AB - BACKGROUND: Prostate specific antigen (PSA) has been used as a screening test for the early detection of prostate cancer (PC) for many years. Although the introduction of PSA test led to a considerable increase in reported prostate cancer cases, there is still some controversy over the sensitivity and specificity of this marker in distinguishing PC patients from those with benign prostate hyperplasia (BPH), the most common benign prostate condition. OBJECTIVE: An attempt is made to elucidate if the plasma level of Interleukin 8 (IL-8) could be used effectively as a marker for the detection of prostate cancer. METHODS: Plasma levels of IL-8 and PSA were measured in two groups of 40 BPH and PC patients using enzyme-linked immunosorbent (ELISA) and radioimmunoassay (RIA) techniques, respectively. In addition IL-8 levels in PC3 and DU145 cell line supernatants were measured by ELISA technique. RESULTS: The concentration of IL-8 in the plasma of PC patients was not significantly higher than the BPH subjects. Although, a correlation between plasma IL-8 concentration and the Gleason score of PC patients was found, no indicated correlation was detected between the concentration of IL-8 or PSA and age of the patients in both groups. DU145 and PC3 cell lines produced and secreted IL-8 in the media. CONCLUSION: Data of this investigation collectively conclude no correlation between IL-8 concentration in PC and BPH patients. PMID- 19561379 TI - Association of HLA-DQA1*0101/2 and DQB1*0502 with myasthenia gravis in southern Iranian patients. AB - BACKGROUND: Myasthenia gravis is an autoimmune disorder of neuromuscular junction characterized by skeletal muscle weakness and fatigability. Different genes may control the induction and clinical presentation of this disease. Various HLA alleles are reported as predisposing or protective genetic elements in myasthenia gravis. OBJECTIVE: The aim of this study was to investigate the probable association between HLA-DQ alleles and myasthenia gravis in southern Iranian patients. METHODS: HLA-DQA1 and DQB1 alleles were determined in 104 sporadic patients with myasthenia gravis using polymerase chain reaction - restriction fragment length polymorphism method and the results were compared to 816 healthy controls. RESULTS: HLA-DQA1*0101/2 (39.4%) and DQB1*0502 (21.6%) were the most frequent alleles in southern Iranian patients with myasthenia gravis. These alleles revealed positive associations with the disease with relative risks of 1.69 and 2.41, respectively. The most common haplotype was DQA1*0101/2-DQB1*0502 in these patients. CONCLUSION: According to the results of this study, DQA1*0101/2 and DQB1*0502 alleles might be considered as predisposing genetic factors to myasthenia gravis while DQA1*0501, DQB1*0301 and *0602/3 show protective roles against this disease. PMID- 19561380 TI - Antiphospholipid syndrome presenting with superior vena cava thrombosis. PMID- 19561381 TI - Hydrodynamics-based transfection of pancreatic duodenal homeobox 1 DNA improves hyperglycemia and is associated with limited complications in diabetic mice. AB - The biohazards caused by the viral delivery of pancreatic duodenal homeobox gene 1 (Pdx1) to the murine liver limits its application. We aimed to evaluate the feasibility of hydrodynamics-based transfection (HBT) with Pdx1 in improving hyperglycemia. Murine hepatocellular carcinoma (Hepa1-6) cells were transfected with the Pdx1-expressing plasmid, pcDNA3.1/V5-His A (pcDNA)-Pdx1. Hepatic delivery of pcDNA-Pdx1 or pcDNA in streptozocin- induced diabetic mice was achieved by HBT. The sequential serum glucose and alanine aminotransferase (ALT) levels were assessed. On the 3(rd) day after transfection, the transfection efficiency in the Hepa1-6 cells and the mice livers was 5% and 0.35 %, respectively. At 1 wk after HBT, asides from hepatic expression of insulin, the diabetic mice transfected with pcDNA-Pdx1 had a significantly lower sugar (211 +/ 61.6 vs. 413 +/- 62 mg/dL; p = 0.002) level than those transfected with pcDNA; however, the difference diminished afterward. No significant difference in the ALT levels was observed between the 2 groups. No mortality was noted in the mice transfected with pcDNA-Pdx1. The hypoglycemic effect of Pdx1 delivered by HBT was transient and associated with negligible complications. In studies on the short term biological effects of Pdx1 in vivo, HBT is a potential alternative to viral delivery of Pdx1 to the murine liver. PMID- 19561382 TI - The sensitivity of the method used to detect atrial fibrillation in population studies affects group-specific prevalence estimates: ethnic and regional distribution of atrial fibrillation in the REGARDS study. AB - BACKGROUND: Among African-Americans, and in southern US states, the rates of stroke are high but the reported prevalences of atrial fibrillation (AF) are low. We hypothesized that the reported ethnic and regional distributions of AF are affected by the sensitivity of the methods that were used to detect AF in previous reports. METHODS: A total of 18,833 black and white participants from the US national REasons For Geographic And Racial Differences In Stroke (REGARDS) study were included in this analysis. Levels of sensitivity to detect AF, from least to most sensitive, were created for combinations of self-report (SR) and ECG methods, as follows: (1) SR plus ECG, (2) ECG alone, (3) SR alone, and (4) SR or ECG. Geographic regions were dichotomized as Stroke Belt (the southern US states) and non-Stroke Belt. Logistic regression analysis estimated the odd ratios of AF associated with the Stroke Belt and black ethnicity for each diagnostic combination. RESULTS: Residence in the Stroke Belt was significantly associated with AF when diagnosed by SR plus ECG (multivariable-adjusted OR, 0.66; 95% CI, 0.47 to 0.92), but not when diagnosed with SR or ECG (OR, 0.95; 95% CI, 0.85 to 1.06). Similarly, for the 4 methods used to detect AF, the strength of the association between black ethnicity and AF progressively decreased with increasing test sensitivity (ORs: 0.20, 0.40, 0.70, 0.71, respectively). CONCLUSIONS: The association of AF with residence in the Stroke Belt and black ethnicity was inversely related to the sensitivity of the method used to detect AF: as test sensitivity increased, the association became attenuated. This may partially explain the lower reported prevalence of AF in populations and regions with higher stroke rates. PMID- 19561383 TI - Hepatitis A surveillance and vaccine use in China from 1990 through 2007. AB - BACKGROUND: Hepatitis A vaccines have been highly effective in preventing hepatitis A. To investigate the epidemiology of hepatitis A in China after hepatitis A vaccine became available, we reviewed reported cases of hepatitis A and the use of hepatitis A vaccine in China during the period from 1990 through 2007. METHODS: Data from the National Notifiable Disease Reporting System from 1990 to 2007 and the Emergency Events Reporting System from 2004 to 2007 were reviewed and epidemiologic characteristics analyzed. Hepatitis A vaccine distribution between 1992 and 2007 was also reviewed. RESULTS: The incidence of hepatitis A has declined by 90% since 1990, from 56 to 5.9 per 10(5)/year. Declines in age-specific incidence were seen in all age groups, most dramatically among children younger than 10 years. Disease incidence still varies substantially: poorer western provinces have had the highest incidences since 2000. In high-incidence provinces, children younger than 10 years continue to have a high disease incidence. Only 50% of cases were laboratory-confirmed, and only 3% occurred in reported local outbreaks. Over 156 million doses of hepatitis A vaccine have been distributed since 1992, and use has continued to increase since 2003. CONCLUSIONS: Incidence of hepatitis A has decreased in all age groups, likely due to changing socioeconomic conditions and increasing hepatitis A vaccine use. Nevertheless, western populations remain at high risk, with transmission predominantly occurring among children. The epidemiology of hepatitis A transmission is not well understood. Improved surveillance with better laboratory confirmation is needed to monitor the impact of universal hepatitis A vaccination of young children; this strategy began to be implemented in 2008. PMID- 19561384 TI - Plantago ovata consumption and colorectal mortality in Spain, 1995-2000. AB - BACKGROUND: Consumption of Plantago ovata may protect against colorectal cancer. To test this hypothesis, an ecological study was performed to determine mortality rates and distribution of colorectal cancer, and the consumption and distribution of P ovata, in different provinces in Spain. The putative association between P ovata consumption and mortality from colorectal cancer was then evaluated. METHODS: We conducted a comparative ecological study of Spanish provinces, with colorectal cancer mortality as the dependent variable and per capita consumption of P ovata by province and year as the independent variable. Associations were analyzed by calculating Spearman's correlation coefficients and a Poisson multiple regression model. RESULTS: Consumption of P ovata tended to be inversely correlated with mortality from colorectal cancer. In the Poisson regression analysis this tendency remained and reached statistical significance for the top quintile of P ovata consumption in the adjusted analysis (P = 0.042). CONCLUSIONS: Our results show an inverse trend between the consumption of P ovata and colorectal cancer mortality. We recommend additional observational studies of individuals, in order to better control confounding factors. PMID- 19561385 TI - The Upcoming 90(th) anniversary of the Tohoku Journal of Experimental Medicine. PMID- 19561386 TI - Identification of amino acid variants in the hepatitis C virus non-structural protein 4A. AB - Chronic infections by hepatitis C virus (HCV) are a major cause of cirrhosis and hepatic cancer. The replication of HCV involves translation and proteolytic processing of polyproteins. The HCV single-stranded RNA encodes a single polyprotein of C/E1/E2/p7/NS2/NS3/NS4A/NS4B/NS5A/NS5B. The structural proteins, C, E1, E2, and p7, arise from the viral polyprotein by host proteases. Cleavage at the non-structural NS2/NS3 junction is performed by the NS2 protease. NS3 forms a complex with NS4A to cleave the rest of the viral polyprotein. The central 12-amino-acid sequence of NS4A, 21-GSVVIVGRIILS-32 (NS4Awt) is a determinant to enhance the NS3 protease activity at the NS5A/5B junction. We found that, from 13 blood donors infected with HCV, one sample showed five amino acid changes in the NS4A central region at V23I, I25C, I30S, L31T, and S32L, and another sample showed three changes at V23I, I25C, and I30V in this region. The other 11 samples showed the NS4Awt sequence. The effect of such amino acid variations on the NS3 proteolytic activity was evaluated in vitro using the central 12-amino-acid NS4Awt sequence with specific changes joined to NS3, and NS5A/5B as a substrate. Our results indicate that the amino acid changes of NS4A at V23I and I25C do not enhance the protease activity of NS3, whereas the amino acid changes at I30S, L31T, and S32L, as well as the NS4Awt sequence, enhance NS3 activity. Our results confirm that protease cofactor, encoded in NS4A, is of major regulatory relevance for the replication cycles of HCV. PMID- 19561387 TI - Diffusion-weighted imaging in the detection of lymph node metastasis in colorectal cancer. AB - Diffusion-weighted imaging (DWI) is a functional technique based on the ability to depict movement of water molecules. The magnitude of water molecule movement is expressed as apparent diffusion coefficient (ADC) value. Its usefulness in the diagnosis of malignant tumors has gained interest. The purpose of this study was to evaluate the usefulness of DWI in detecting lymph node metastases of colorectal cancer. The subjects were 46 consecutive patients (mean age 71.4 +/- 8.7 years) with colorectal cancer, treated by radical surgery from 2006 to 2008. The size of metastatic lymph nodes on DWI was significantly larger than non metastatic lymph nodes (10.3 vs. 7.6 mm). The mean ADC value was significantly lower for metastatic lymph nodes than non-metastatic lymph nodes (1.36 vs. 1.85 x 10(-3) mm(2)/sec). In addition, for evaluation of lymph node metastasis that reflects the primary tumor characteristics, the LN/T ratio (defined as the ratio of lymph node ADC value to the primary tumor ADC value) was calculated. It was significantly lower for metastatic lymph nodes than non-metastatic lymph nodes (1.41 vs. 1.59). Receiver operating characteristic curve analysis revealed that the best performing cutoffs were 8.5 mm for lymph node size, 1.44 x 10(-3) mm(2)/sec for ADC value, and 1.495 for LN/T ratio. Accuracy was significantly greater for lymph nodes with LN/T ratio (78.5%) than for lymph node size (62.0%) or ADC value (74.8%). In conclusion, preoperative DWI, especially the LN/T ratio, is recommended for evaluation of lymph node metastasis in patients with colorectal cancer. PMID- 19561388 TI - Common polymorphisms in the XPD and hOGG1 genes are not associated with the risk of colorectal cancer in a Polish population. AB - Mutations in the DNA repair genes may contribute to the increased risk of cancer, including colorectal cancer. Xeroderma pigmentosum group D (XPD) protein and human homolog of the 8-oxoguanine glycosylase 1 (hOGG1) are involved in nucleotide excision repair and base excision repair, respectively. The XPD and the hOGG1 genes are highly polymorphic, and some of their polymorphisms are associated with several types of cancers. However, there is controversy as to the relationship between their polymorphisms and the risk of colorectal cancer. In the present study, we therefore searched for the association in a Polish population between colorectal cancer and two common polymorphisms: an A --> C transversion in the XPD gene that produces a Lys-to-Gln substitution at codon 751 (the Lys751Gln polymorphism; rs28365048) and a C --> G transversion in the hOGG1 gene resulting in a Ser-to-Cys change at codon 326 (the Ser326Cys polymorphism; rs1052133). Genotypes were determined using peripheral blood lymphocytes of 100 colorectal cancer patients and 100 age-, sex- and ethnicity-matched cancer-free controls by PCR and restriction fragment-length polymorphism analysis. We did not find statistically significant association between each polymorphism and the occurrence of colorectal cancer, and did not observe any relationship between each polymorphism and colorectal cancer progression assessed by node metastasis, tumor size and Duke's stage. Moreover, there was no correlation between combined genotypes of the two polymorphisms and colorectal cancer. Therefore, the Lys751Gln polymorphism of the XPD gene and the Ser326Cys polymorphism of the hOGG1 gene are not associated with colorectal cancer in a Polish population. PMID- 19561389 TI - Improving gait stability in stroke hemiplegic patients with a plastic ankle-foot orthosis. AB - Stroke is the leading cause of long-term disability, and many stroke patients have hemiparesis. Hemiparesis induces ankle-control disturbances and equinovarus deformity, leading to difficulty in walking and an increased risk for falling. Plastic ankle-foot orthosis (PAFO) is frequently prescribed to correct ankle joint alignment and increase walking speed and stride length during ambulation. While several studies have shown that PAFO improves gait parameters, such as stride length and walking speed, in hemiplegic patients, the effect of PAFO on gait stability remains unclear. We quantitatively assessed the effect of PAFO on gait stability in 16 hemiplegic stroke patients (mean age 55.9 +/- 11.8 years; 5 female and 11 male subjects; and 11 hemorrhagic and 5 ischemic stroke) using an ink footprint record. Wearing PAFO significantly improved the stride length, step length on the unaffected and affected sides, step width, walking speed, step frequency and functional ambulation ability. The coefficient of variation (CV), as an index of stability of movement from trial to trial, provides a measure that defines motor skills for a given task. Unaffected-side step-length CV and step width CV were significantly decreased, when using PAFO. Furthermore, the correlation was found only between unaffected-side step length and its CV. The decrease in CV indicates that PAFO improved gait stability. We concluded that in addition to providing a faster gait, PAFO improves gait stability during walking. Gait stability and gait efficiency need to be considered separately in evaluating the effects of ankle-foot orthosis on gait performance in hemiplegic patients. PMID- 19561391 TI - Sequence-based spa typing as a rapid screening method for the areal and nosocomial outbreaks of MRSA. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is the leading cause of nosocomial infection and MRSA outbreaks have become a major problem. Therefore, the rapid and accurate typing of MRSA isolates is important for epidemiological surveys and nosocomial infection control. Pulsed-field gel electrophoresis (PFGE) is considered as the gold standard technique for MRSA typing, because of its high discriminatory power, but its procedure is rather complicated and time-consuming. The spa gene encodes a cell wall component of Staphylococcus aureus protein A, and exhibits polymorphism. Sequencing the spa gene is expected superior to PFGE in speed and data interpretation. In the present study, we evaluated whether spa typing of MRSA is useful for nosocomial outbreak analysis and epidemiological investigations. We analyzed 19 nosocomial outbreak isolates from 4 separate hospitals and 26 isolates from outpatients of Toyama University Hospital. Either PFGE or spa typing revealed a single nosocomial strain that appears unique to each hospital. Indeed, spa typing confirmed the four different strains, but PFGE demonstrated only 3 strains. With the total 45 isolates, PFGE showed 16 different patterns and spa typing showed 12 patterns. Moreover, we were able to analyze the spa gene in about 2 days, from sampling to obtaining the results, whereas it took about 7 days with PFGE. In conclusion, sequence-based spa typing shows comparable sensitivities to PFGE, and is a rapid and easy handling method. The sequence based spa typing can be used as the rapid screening test when MRSA outbreak is suspected in areas and hospitals. PMID- 19561390 TI - Low serum levels of undercarboxylated osteocalcin in postmenopausal osteoporotic women receiving an inhibitor of bone resorption. AB - Osteocalcin, a bone-specific protein synthesized by osteoblasts, undergoes vitamin K-dependent gamma-carboxylation. Undercarboxylated osteocalcin (ucOC) represents inadequately carboxylated osteocalcin, and this fraction increases with vitamin K insufficiency. Alendronate is a bisphosphonate that inhibits bone resorption, thereby increasing bone mineral density (BMD), while also reducing bone formation closely coupled with bone resorption. The aim of this cross sectional study was to evaluate the influence of alendronate on serum levels of ucOC, cross-linked N-telopeptide of type 1 collagen (NTx), a marker of bone resorption, and bone alkaline phosphatase (BAP), a marker of bone formation. Forty-six postmenopausal osteoporotic women were divided into three groups: patients receiving alendronate (5 mg/day or 35 mg/week) for >or= 6 months (n = 29) or < 6 months (n = 7), and patients receiving no medication related to bone metabolism (n = 10). Serum ucOC levels were significantly lower in patients with long-term treatment (p < 0.0001) or short-term treatment (p = 0.0223) than in untreated patients. Serum ucOC levels correlated positively with both BAP (r = 0.695, p < 0.0001) and NTx (r = 0.494, p = 0.0004) in all participants. Since low serum levels of BAP and NTx are associated with decreased levels of bone formation and bone resorption, respectively, these findings suggest that low serum ucOC levels may reflect the suppression of bone turnover. In conclusion, low serum ucOC levels reflect suppressed bone turnover and/or adequate levels of vitamin K in patients receiving an inhibitor of bone resorption. PMID- 19561392 TI - Effective home-based pulmonary rehabilitation in patients with restrictive lung diseases. AB - Patients with chronic obstructive pulmonary disease (COPD) are commonly referred for pulmonary rehabilitation (PR), but the use of PR is not common for patients with restrictive lung disease, neuromuscular diseases, and those who have sustained a severe respiratory illness or undergone thoracic surgery. We investigated the effects of PR in patients with restrictive lung diseases in comparison with COPD patients using a home-based setting. Twenty-six restrictive lung diseases patients and 40 COPD patients who had a Medical Research Council (MRC) dyspnea score >or= 2, a clinically stable condition, and who had completed a 6-month PR program, were enrolled in the present study. The definition of restrictive lung disease was a forced vital capacity (FVC) of 70%. Our PR consisted of breathing retraining, exercise training, respiratory muscle stretching calisthenics, level walking, inspiratory and expiratory muscle exercises, and a monthly education program. Patients were strongly instructed to practice this program daily at home, and were supervised by a respiratory therapist every 2 weeks in our hospital. Patients with restrictive lung diseases showed the significant increases in inspiratory and expiratory muscle forces, the 6-minute walking distance, the Chronic Respiratory Disease Questionnaire and the Short-Form 36, and decreased MRC scores after 6 months. In conclusion, our home based PR improves respiratory muscle forces, exercise tolerance, health-related quality of life, and the perception of dyspnea in patients with restrictive lung disease to the same extent as in COPD patients. PMID- 19561393 TI - Impaired postnatal growth of infants prenatally exposed to cigarette smoking. AB - Most of the previous studies have shown a significant inverse relationship between smoking during pregnancy and weight, height and head circumference of infants at birth, but there is limited literature that assesses the head circumference measures of infants of smoker mothers in postnatal follow-up. The aim of this study was to assess the effects of maternal smoking and passive smoking during pregnancy on postnatal anthropometric measures of infants. Infants were divided into 3 groups: infants of smokers (n = 48), passive smokers (n = 57) and nonsmokers (n = 54), and were evaluated for their weight, height and head circumference at birth, 3 months and 6 months of age. Infants of smokers showed significant weight and head circumference deficits at birth compared to nonsmokers' infants (p < 0.05 and p < 0.001, respectively). At 6 months of age, infants of smokers continued to show significant deficits in all 3 measures compared to nonsmokers' infants (p < 0.001 for each), and infants of passive smokers showed only marginal decreases. Moreover, the weight and height growth velocities of the smokers' infants remained deficient, whereas their growth velocity of the head circumferences increased from birth up to 6 months and reached the growth velocity of the nonsmokers' infants. Infants of passive smokers showed a complete catch-up growth at 6 months. This study indicates that smoking during pregnancy results in serious deficits in infants' growth even after birth. Therefore, it is essential to inform smoker women before pregnancy the possible growth retardation of infants. PMID- 19561394 TI - Inequity in dental care utilization in the Indonesian population with a self assessed need for dental treatment. AB - Dental caries is one of the most prevalent infectious diseases in Indonesia. More than half of all cases are left untreated. This may be due to inequity in the use of dental care that is caused by economic and geographic barriers. Therefore, the objective of this study was to provide evidence of socioeconomic-related inequality and horizontal inequity in dental care utilization among Indonesian adults who reported having had dental problems and to describe the sources of any identified inequality. We used secondary cross-sectional data from the Indonesian National Socio-Economic Survey 2004 and from the Indonesian Medical Council. Respondents included individuals at least 15 years old who reported having had dental care needs within a one-month recall period (N = 20,718). A concentration curve and a concentration index were employed to describe the extent of inequality. A horizontal inequity index was applied to identify inequity. A decomposition method was used to describe the sources of inequality. The concentration curve indicated a slightly pro-rich inequality in dental care utilization. The concentration index showed a significant concentration of dental care utilization among groups with higher socioeconomic status (SES). The horizontal inequity index illustrated higher unmet dental care needs among lower SES groups. Decomposition revealed that higher SES, urban Java Island residency, and insurance coverage were positively associated with the likelihood of dental care utilization. This study concludes that the expansion of health insurance, especially targeted at low SES groups, and a regionally equitable distribution of dentists may reduce economic and geographic barriers to dental care in the future. PMID- 19561395 TI - Determinants of the utilization of dental services in a community-dwelling elderly Japanese population. AB - Oral health care is not only an effective strategy for the prevention, early diagnosis, and treatment of orofacial disease and disorders, but also an essential component of general health promotion programs. The current study aimed to identify the determinants of the utilization of dental services in an elderly population in Japan. A community-based comprehensive geriatric assessment, including the measurements of physical, mental, and social functioning, was conducted among elderly people aged >or= 70 years residing in a suburban area of Sendai, Japan. Oral health status and functioning, their impact on the quality of life, and dental utilization were also surveyed. Of the 1,170 participants, 418 subjects who had specific treatment needs for dental problems and reported irregular dental attendance were recommended a dental visit, and 1 year later, their compliance with the recommendation was assessed by using questionnaire. Multiple logistic regression analysis demonstrated that a higher number of remaining teeth and the use of removable dentures were significant predictors of dental utilization within 1 year. Regular utilization was associated with a higher number of remaining teeth, younger age, presence of systemic disease, absence of depressive symptoms, and higher educational attainment. Conversely, non-compliance with the treatment recommendations was associated with fewer remaining teeth, smoking, and non-utilization of dental services during the previous year. The differences in the determinants of dental attendance behavior, which may be partially associated with the insurance coverage for dental services, suggest the need for specific strategies for oral health promotion for different behavior of dental utilization. PMID- 19561396 TI - Resveratrol, a natural antioxidant, attenuates intestinal ischemia/reperfusion injury in rats. AB - The intestine is highly susceptible to ischemia/reperfusion (I/R) injury. Splanchnic ischemia is the initial event that releases injurious factors, leading to systemic disorders with high morbidity and mortality. Oxidative stress mediators are believed to contribute to the intestinal I/R injury. Resveratrol, a polyphenol found in grapes, is shown to be a strong antioxidant in various tissues, with a property of an estrogen-receptor agonist. Therefore, we investigated the effects of resveratrol on oxidative injury in the intestine. Female Wistar rats were randomly allocated into four groups (n = 8, each). The sham group was only subjected to surgical procedures, while other animals were subjected to intestinal ischemia (60 min) and subsequent reperfusion (60 min). One group received resveratrol (15 mg/kg, 0.3 ml/day intraperitoneally) for both 5 days before surgery and 15 min before ischemia, while the other was treated intraperitoneally with 0.5% ethyl alcohol as vehicle (0.3 ml/day). In the I/R rat intestines, we detected severe tissue injuries (p < 0.001), the significant increases in the tissue levels of malondialdehyde (MDA), nitric oxide (NO), and myeloperoxidase (MPO) (p < 0.001), and the decrease in superoxide dismutase (SOD) activity (p < 0.001), compared to the sham control. Resveratrol significantly ameliorated the intestinal injury, decreased MDA, NO and MPO levels to the sham control levels, and decreased bacterial translocation in mesentery lymph nodes, liver and spleen (p < 0.001). Resveratrol also restored the SOD activity. These results suggest that resveratrol could protect intestinal tissue against I/R injury with its potent antioxidant properties. PMID- 19561397 TI - Interleukin-1beta promotes the expression of monocyte chemoattractant protein-1 in human aorta smooth muscle cells via multiple signaling pathways. AB - Monocyte chemoattractant protein-1 (MCP1) plays a key role in monocyte/macrophage infiltration to the sub-endothelial space of the blood vessel wall, which is a critical initial step in atherosclerosis. In this study, we examined the intracellular signaling pathway of IL-1beta-induced MCP1 expression using various chemical inhibitors. The pretreatment of a phosphatidylcholine (PC)-specific PLC (PC-PLC) inhibitor (D609), PKC inhibitors, or an NF-kapaB inhibitor completely suppressed the IL-1beta-induced MCP1 expression through blocking NF-gammaB translocation to the nucleus. Pretreatment with inhibitors of tyrosine kinase or PLD partially suppressed MCP1 expression and failed to block nuclear NF-kappaB translocation. These results suggest that IL-1beta induces MCP1 expression through activation of NF-kappaB via the PC-PLC/PKC signaling pathway. PMID- 19561399 TI - Alpha-tocopheryl succinate potentiates the paclitaxel-induced apoptosis through enforced caspase 8 activation in human H460 lung cancer cells. AB - Paclitaxel is one of the chemotherapeutic drugs widely used for the treatment of nonsmall cell lung cancer (NSCLC) patients. Here, we tested the ability of alpha tocopheryl succinate (TOS), another promising anticancer agent, to enhance the paclitaxel response in NSCLC cells. We found that sub-apoptotic doses of TOS greatly enhanced paclitaxel-induced growth suppression and apoptosis in the human H460 NSCLC cell lines. Our data revealed that this was accounted for primarily by an augmented cleavage of poly(ADP-ribose) polymerase (PARP) and enhanced activation of caspase-8. Pretreatment with z-VAD-FMK (a pan-caspase inhibitor) or z-IETD-FMK (a caspase-8 inhibitor) blocked TOS/paclitaxel cotreatment-induced PARP cleavage and apoptosis, suggesting that TOS potentiates the paclitaxel induced apoptosis through enforced caspase 8 activation in H460 cells. Furthermore, the growth suppression effect of TOS/paclitaxel combination on human H460, A549 and H358 NSCLC cell lines were synergistic. Our observations indicate that combination of paclitaxel and TOS may offer a novel therapeutic strategy for improving paclitaxel drug efficacy in NSCLC patient therapy as well as for potentially lowering the toxic side effects of paclitaxel through reduced drug dosage. PMID- 19561398 TI - Comparative study of synonymous codon usage variations between the nucleocapsid and spike genes of coronavirus, and C-type lectin domain genes of human and mouse. AB - Coronaviruses (CoVs) are single-stranded RNA viruses which contain the largest RNA genomes, and severe acute respiratory syndrome coronavirus (SARS-CoV), a newly found group 2 CoV, emerged as infectious disease with high mortality rate. In this study, we compared the synonymous codon usage patterns between the nucleocapsid and spike genes of CoVs, and C-type lectin domain (CTLD) genes of human and mouse on the codon basis. Findings indicate that the nucleocapsid genes of CoVs were affected from the synonymous codon usage bias than spike genes, and the CTLDs of human and mouse partially overlapped with the nucleocapsid genes of CoVs. In addition, we observed that CTLDs which showed the similar relative synonymous codon usage (RSCU) patterns with CoVs were commonly derived from the human chromosome 12, and mouse chromosome 6 and 12, suggesting that there might be a specific genomic region or chromosomes which show a more similar synonymous codon usage pattern with viral genes. Our findings contribute to developing the codon-optimization method in DNA vaccines, and further study is needed to determine a specific correlation between the codon usage patterns and the chromosomal locations in higher organisms. PMID- 19561400 TI - The adaptor protein LAD/TSAd mediates laminin-dependent T cell migration via association with the 67 kDa laminin binding protein. AB - The adaptor protein, LAD/TSAd, plays essential roles in T cell activation. To further understand the functions of this protein, we performed yeast two-hybrid screening using TSAd as bait and identified 67 kDa laminin binding protein (LBP) as the interacting partner. Subsequently, TSAd-LBP interaction was confirmed in D1.1 T cell line. Upon costimulation by T cell receptor (TCR) plus laminin crosslinking or TCR plus integrin alpha6 crosslinking, LBP was coimmunoprecipitated with TSAd. Moreover, TCR plus laminin costimulation dependent T cell migration was enhanced in D1.1 T cells overexpressing TSAd but was disrupted in D1.1 cells overexpressing dominant negative form of TSAd or TSAd shRNA. These data show that, upon TCR plus integrin costimulation, TSAd associates with LBP and mediates T lymphocyte migration. PMID- 19561401 TI - Triptolide downregulates Rac1 and the JAK/STAT3 pathway and inhibits colitis related colon cancer progression. AB - Triptolide, a diterpenoid triepoxide from the traditional Chinese medicinal herb Tripterygium wilfordii Hook. f., is a potential treatment for autoimmune diseases as well a possible anti-tumor agent. It inhibits proliferation of colorectal cancer cells in vitro and in vivo. In this study, its ability to block progress of colitis to colon cancer, and its molecular mechanism of action are investigated. A mouse model for colitis-induced colorectal cancer was used to test the effect of triptolide on cancer progression. Treatment of mice with triptolide decreased the incidence of colon cancer formation, and increased survival rate. Moreover, triptolide decreased the incidence of tumors in nude mice inoculated with cultured colon cancer cells dose-dependently. In vitro, triptolide inhibited the proliferation, migration and colony formation of colon cancer cells. Secretion of IL6 and levels of JAK1, IL6R and phosphorylated STAT3 were all reduced by triptolide treatment. Triptolide prohibited Rac1 activity and blocked cyclin D1 and CDK4 expression, leading to G1 arrest. Triptolide interrupted the IL6R-JAK/STAT pathway that is crucial for cell proliferation, survival, and inflammation. This suggests that triptolide might be a candidate for prevention of colitis induced colon cancer because it reduces inflammation and prevents tumor formation and development. PMID- 19561402 TI - Sequential evolution of IL-17 responses in the early period of allograft rejection. AB - In addition to CD4+CD25+Foxp3+ regulatory T (T(reg)) cells which protect against autoimmune tissue injury, IL-17-producing CD4+ T (T(h)17) cells have been recently described and shown to play a crucial role in autoimmune injury. It appears that there is a reciprocal developmental pathway between T(h)17 and T(reg) cells. Although IL-17 is known to be associated with allograft rejection, the cellular source of IL-17 and the nature of T(h)17 in the context of allograft rejection remain unknown. In the current study, the dynamics of T(reg) and IL-17 producing cells after syngeneic and allogeneic transplantation were examined using a wild-type murine cardiac transplantation model. Ly6G+ cells were found to produce IL-17 during the early postoperative period and CD8+ as well as CD4+ T cells were also found to produce IL-17 during alloimmune response. Graft infiltrating Ly6G+, CD4+, and even CD8+ cells were found to express IL-17 highly compared to those in spleen. Although the frequencies of T(h)17 and T(reg) were found to gradually increase in both syngeneic and allogeneic recipients, T(h)17/T(reg) ratios were significantly higher in recipients with allograft rejection than in syngeneic recipients. In conclusion, IL-17 is produced by neutrophils during the early postoperative period and subsequently by T(h)17 and CD8+ T cells during allograft rejection. T(h)17/T(reg) imbalance is associated with the development of allograft rejection. This study would provide basic information on T(h)17 biology for future investigation in the field of transplantation. PMID- 19561403 TI - Negative feedback regulation of Wnt signaling by Gbetagamma-mediated reduction of Dishevelled. AB - Wnt signaling is known to be important for diverse embryonic and post-natal cellular events and be regulated by the proteins Dishevelled and Axin. Although Dishevelled is activated by Wnt and involved in signal transduction, it is not clear how Dishevelled-mediated signaling is turned off. We report that guanine nucleotide binding protein beta 2 (Gnb2; Gbeta2) bound to Axin and Gbeta2 inhibited Wnt mediated reporter activity. The inhibition involved reduction of the level of Dishevelled, and the Gbeta2gamma2 mediated reduction of Dishevelled was countered by increased expression of Axin. Consistent with these effects in HEK293T cells, injection of Gbeta2gamma2 into Xenopus embryos inhibited the formation of secondary axes induced either by XWnt8 or Dishevelled, but not by beta-catenin. The DEP domain of Dishevelled is necessary for both interaction with Gbeta2gamma2 and subsequent degradation of Dishevelled via the lysosomal pathway. Signaling induced by Gbeta2gamma2 is required because a mutant of Gbeta2, Gbeta2 (W332A) with lower signaling activity, had reduced ability to downregulate the level of Dishevelled. Activation of Wnt signaling by either of two methods, increased Frizzled signaling or transient transfection of Wnt, also led to increased degradation of Dishevelled and the induced Dishevelled loss is dependent on Gbeta1 and Gbeta2. Other studies with agents that interfere with PLC action and calcium signaling suggested that loss of Dishevelled is mediated through the following pathway: Wnt/Frizzled-->Gbetagamma-->PLC-->Ca(+2)/PKC signaling. Together the evidence suggests a novel negative feedback mechanism in which Gbeta2gamma2 inhibits Wnt signaling by degradation of Dishevelled. PMID- 19561405 TI - Reduced-intensity conditioning for allogeneic stem cell transplantation: 10 years later. PMID- 19561404 TI - Legionella lipoprotein activates toll-like receptor 2 and induces cytokine production and expression of costimulatory molecules in peritoneal macrophages. AB - Legionella bacterium, an intracellular pathogen of mononuclear phagocytes, causes acute fatal pneumonia, especially in patients with impaired cellular immune responses. Until recently, however, the toll-like receptor (TLR) engagement of bacterial proteins derived from Legionella is uncertain. We previously showed that a 19-kDa highly conserved peptidoglycan-associated lipoprotein (PAL) of Legionella pneumophila induced the PAL-specific B cell and T cell responses in mice. In this study, we observed that the rPAL antigen of L. pneumophila, as an effector molecule, activated murine macrophages via TLR2 and produced proinflammatory cytokines such as IL-6 and TNF-alpha. In both BALB/c and TLR4 deficient C3H/HeJ mice, pretreatment of macrophages with anti-TLR2 mAb showed severely impaired cytokine production in response to the rPAL. In addition, in vitro the rPAL treatment increased the cell surface expression of CD40, CD80, CD86 and MHC I/II molecules. We further showed that the synthetic CpG oligodeoxynucleotides (CpG ODN) coadministered with the rPAL enhanced IL-12 and IL-6 production and expression of CD40, CD80 and MHC II compared to the rPAL treatment alone. In conclusions, these results indicate that Legionella PAL might activate macrophages via a TLR2-dependent mechanism which thus induce cytokine production and expression of costimulatory and MHC molecules. PMID- 19561407 TI - Low-dose total body irradiation-based regimens as a preparative regimen for allogeneic haematopoietic cell transplantation in acute myelogenous leukaemia. AB - Although much progress has been made in understanding the molecular basis of acute myeloid leukaemia (AML), this has not yet led to major improvements in the overall survival of patients. In particular, the treatment of elderly patients with AML remains one of the major challenges in haematology. Despite increases in complete remission rates, relapse remains a major obstacle and the major determinant of overall survival. Allogeneic stem cell transplantation (SCT) is the most efficient antileukaemic treatment for patients with AML, but eligibility for the treatment was confined for a long time to younger patients. More than 10 years ago, SCT protocols were initiated with reduced-intensity conditioning (RIC) rather than ablative chemoradiotherapy, with the intention of inducing a graft versus-leukaemia effect also in elderly patients and patients with concomitant diseases. Operationally, all protocols below the conventional preparative regimens are referred to as RIC. Low-dose total body irradiation-based protocols result in minimal myelosuppression and are among the most popular. After extensive phase I and phase II studies, associated problems of graft rejection have been largely resolved and transplant-related mortality (TRM) evaluated in more than 3000 patients. TRM does not currently exceed 10-12% in related and 20% in unrelated SCT even in patients up to the age of 75 years, so that relapse after transplantation remains the major problem. A number of strategies for decreasing relapse rates has been developed. The most promising approach consists of monitoring CD34+ donor cell chimerism after transplantation. This has led to decreases in the relapse rate over the past few years. Randomized studies are now being initiated to define the role of SCT in the treatment of elderly patients with AML. PMID- 19561406 TI - Reduced-toxicity conditioning therapy with allogeneic stem cell transplantation for acute leukemia. AB - We hypothesized that standardized systemic drug delivery would improve treatment safety and provide better leukemia control. We therefore developed an intravenous busulfan formulation and combined it with fludarabine instead of cyclophosphamide in preparation for allogeneic stem cell transplantation (alloSCT). We used a Bayesian method to compare the outcomes of 67 acute myeloid leukemia (AML)/myelodysplastic syndrome (MDS) patients who received intravenous busulfan cyclophosphamide (BuCy2) with 148 subsequent AML/MDS patients who received busulfan-fludarabine (Bu-Flu). The groups had comparable pretreatment characteristics, except that the Bu-Flu patients were older, more often had unrelated donors and had a shorter follow-up. A greatly improved outcome in the Bu-Flu group is unlikely to be explained by improved supportive care during this time period. Overall, the data support replacing BuCy2 with or without antithymocyte globulin (ATG) with Bu-Flu with or without rabbit-ATG for AML or MDS. We further suggest that the high level of safety and fast recovery from conditioning therapy-related side effects after the Bu-Flu regimen makes it a suitable platform technology for testing additional adjuncts for improved posttransplant immune recovery and long-term disease control in patients who are at high risk of rapidly recurrent disease after alloSCT. The extremely low one year treatment-related mortality as well as high overall and event-free survival of patients in the Bu-Flu group indicate that it is time to revisit the value of alloSCT compared with conventional maintenance chemotherapy for patients in first complete remission of AML/MDS. PMID- 19561408 TI - Reduced-intensity conditioning allogeneic stem cell transplantation with donor T cell depletion using alemtuzumab added to the graft ('Campath in the bag'). AB - Reduced-intensity conditioning (RIC) has allowed the use of allogeneic stem cell transplantation (alloSCT) for haematological malignancies in elderly patients. A major problem of this type of transplantation is the high incidence of persisting chronic graft-versus-host disease (GvHD), leading to increased morbidity and mortality. The inclusion of alemtuzumab added to the graft ('Campath in the bag') for donor T-cell depletion offers an easy procedure to diminish the incidence of GvHD. Good engraftment is observed in most patients, whereas almost no GvHD is observed after transplantation. Most patients become mixed chimeric after transplantation, requiring donor lymphocyte infusion for conversion to full donor chimerism. Although subsequent acute and chronic GvHD is observed in 50-60% of patients, it is responsive to therapy in many patients, resulting in a low incidence of persisting chronic GvHD. AlloSCT with RIC and alemtuzumab-induced T cell depletion offers a suitable platform for the investigation of novel cellular immunotherapy. PMID- 19561409 TI - Total lymphoid irradiation for graft-versus-host disease protection. AB - PURPOSE: The immunosuppressive effects of total lymphoid irradiation (TLI) for protection against graft-versus-host disease (GvHD) after allogeneic bone marrow transplantation have been studied for years in animal models. In preclinical models of bone marrow transplantation non-myeloablative TLI conditioning protects against GvHD by skewing host T-cell subsets to favor regulatory natural killer T cells that suppress GvHD by polarizing donor T cells towards secretion of non inflammatory cytokines such as IL-4. These preclinical models have recently been adapted to human transplantation. RECENT FINDINGS: Patients receiving allogeneic hematopoietic cell transplantation for hematological malignancies conditioned with TLI and depletive T-cell antibodies showed sustained donor chimerism, a reduced incidence of acute GvHD yet retained anti-tumor activity. As in the preclinical models, the low incidence of GvHD is associated with increased IL-4 secretion by chimeric donor T cells. SUMMARY: This review summarizes the evolution of the preclinical TLI protocols and their recent translation to clinical trials, and discusses the mechanisms involved in protection from GvHD and the induction of tolerance following mixed chimerism. PMID- 19561410 TI - Reduced-intensity conditioning transplantation in myeloid malignancies. AB - The development of non-myeloablative and reduced-intensity conditioning regimens has enabled older or medically infirm patients with myeloid malignancies to be treated with allogeneic hematopoietic cell transplantation (HCT). The regimens are sufficiently immunosuppressive to allow engraftment of allogeneic cells and they rely largely on graft-versus-leukemia effects rather than high-dose cytotoxic therapy to eliminate malignant cells. Overall 2-5-year survivals after allogeneic HCT in older patients with acute myeloid leukemia (AML) or myelodysplastic syndromes (MDS) have ranged from 25% to 64%. Outcomes were comparable for human leukocyte antigen-matched related and unrelated grafts. The best results were seen in patients transplanted in the first or second remission. Relapse and progressive disease continue to be problems, particularly in patients with large tumor burdens at the time of HCT. Reduction of the tumor burden before HCT with targeted therapy such as radiolabelled anti-CD45 antibody may improve the outcome. Despite still existing problems, early results in elderly patients with AML/MDS have been encouraging. PMID- 19561411 TI - Reduced-intensity conditioning regimens before unrelated cord blood transplantation in adults with acute leukaemia and other haematological malignancies. AB - Cord blood is an unlimited source of haematopoietic stem cells for allogeneic haematopoietic stem cell transplants. During the past 5 years, the number of adults transplanted with cord blood cells from unrelated donors has exceeded the number of transplants in children, as a result of better definitions of cord blood unit choice, an increased number of cord blood units available for transplantation worldwide, comparable results of unrelated cord blood transplantation (UCBT) with human leukocyte antigen-matched unrelated bone marrow transplantation, the use of double cord blood transplantation and the use of UCBT after a reduced-intensity conditioning (RIC) regimen. In spite of the encouraging results of RIC UCBT in single-centre studies, the number of patients given this strategy is still limited and follow-up is still too short to draw definitive conclusions. Moreover, many questions remain to be answered such as: (1) the type of patients and disease populations that may benefit most from this strategy; (2) the best conditioning regimen to use; (3) the criteria of cord blood choice in this setting; and (4) factors predictive of outcomes after RIC UCBT. This paper will summarize some recent results of RIC UCBT for adults with haematological malignancies. PMID- 19561412 TI - Allogeneic stem cell transplantation after reduced-intensity conditioning for acute myeloid leukaemia: impact of chronic graft-versus-host disease. AB - The antineoplastic effect of allogeneic stem cell transplantation after reduced intensity conditioning (RIC) relies on the graft-versus-tumour (GvT) reaction. GvT is closely linked to the development of graft-versus-host disease (GvHD). The incidence of acute GvHD after RIC seems lower than after myeloablative conditioning (MAC), whereas the incidence of chronic GvHD after RIC seems similar to after MAC. The results of RIC for acute myeloid leukaemia show a non-relapse mortality of approximately 15% at one year, a relapse incidence of approximately 40% after a median of 4-6 months, translating into overall and disease-free survival rates of 40-60%. The factors associated with improved outcome in most studies are the stage of the disease at transplantation, age and the development of chronic GvHD (and thus GvT). In a recent report, chronic GvHD was the most important factor associated with prolonged survival. Future efforts should be directed at aiming to decrease relapse rates. For this purpose, an adequate identification of high-risk patients, close monitoring of minimal residual disease after the procedure, and the use of antineoplastic drugs or immunotherapy may be of help. PMID- 19561414 TI - First-line treatment of Philadelphia chromosome-positive acute lymphoblastic leukaemia in adults. AB - The tyrosine kinase inhibitor (TKI) imatinib has become an integral part of front line therapy for Philadelphia chromosome-positive acute lymphoblastic leukaemia, with remission rates exceeding 90% irrespective of whether imatinib is given alone or combined with chemotherapy. Treatment outcome with imatinib-based regimens has improved compared with historic controls, but most patients who do not undergo allogeneic stem cell transplantation (SCT) eventually relapse. Second generation TKI, e.g. dasatinib and nilotinib, show activity against most of the bcr-abl tyrosine kinase domain mutations involved in acquired imatinib resistance, but clinical benefit is generally short lived. Accordingly, SCT in first complete response is considered to be the best curative option. Strategies to improve outcome in patients ineligible for transplantation as well as after SCT include front-line treatment with more effective TKI to increase molecular response rates. Following SCT, the pre-emptive use of imatinib appears to reduce the relapse rate. Novel immunotherapeutic interventions and combinations of TKI are also being explored. PMID- 19561413 TI - Pathophysiology and management of graft-versus-host disease in the era of reduced intensity conditioning. AB - Both indirect and direct effects contribute to changes in the clinical presentation of graft-versus-host disease (GvHD) after reduced-intensity conditioning (RIC) compared with standard intensity regimens. A delay in acute GvHD after RIC may be explained by reduced conditioning-related inflammation and by altered allostimulatory capacities of recipient antigen-presenting cells (APC). A higher frequency of chronic GvHD results from the almost exclusive use of peripheral blood stem cells, the absence of tolerance induction by current regimens for prophylactic immunosuppression and modified kinetics in the replacement of recipient APC. Established acute and chronic GvHD requires standard treatment irrespective of the type of conditioning. To improve long-term outcome in GvHD, pre-emptive strategies or more selective approaches aimed at immunomodulation rather than immunosuppression are needed. PMID- 19561415 TI - Maximum supportive care, standard conditioning and allogeneic stem cell transplantation for elderly patients with acute myelogenous leukemia. AB - Dose-intense conditioning (DIC) (myeloablative) regimens for allogeneic stem cell transplantation (alloSCT) were previously avoided in patients with acute myelogenous leukemia aged more than 55 years because of the fear of excessive morbidity and mortality. The introduction of reduced-intensity conditioning (RIC) alloSCT led to their increasing use in these patients because of their reduced toxicity. Significant disadvantages remain, however, including the late establishment of a posttransplant graft-versus-leukemia effect and an overrepresentation of poor prognostic factors in elderly patients, resulting in the risk of early relapse/progression before the graft-versus-leukemia effect being disproportionally large. Preliminary results suggest that DIC with maximum prophylaxis and support is safe for elderly patients up to age 70 years. We hypothesize that DIC may be important for the early control of leukemia in elderly patients, and that prospective, randomized trials comparing DIC and RIC based transplants should be carried out, with the expectation that early transplant-related mortality will be no different. PMID- 19561421 TI - Meta-analysis of trials of streptococcal throat treatment programs to prevent rheumatic fever. AB - BACKGROUND: Rheumatic fever (RF) is the commonest cause of pediatric heart disease globally. Penicillin for streptococcal pharyngitis prevents RF. Inequitable access to health care persists. PURPOSE: To investigate RF prevention by treating streptococcal pharyngitis in school- and/or community-based programs. DATA SOURCES: Medline, Old Medline, the Cochrane Library, DARE, Central, NHS, EED, NICE, NRMC, Clinical Evidence, CDC website, PubMed, and reference lists of retrieved articles. Known researchers in the field were contacted where possible. METHODS: Randomized, controlled trials or trials of before/after design examining treatment of sore throats in schools or communities with RF as an outcome where data were able to be pooled for analysis. Two authors examined titles, abstracts, selected articles, and extracted data. Disagreements were resolved by consensus. QUANTITATIVE ANALYSIS TOOL: Review Manager version 4.2 to assess pooled relative risks and 95% confidence intervals. DATA SYNTHESIS: Six studies (of 677 screened) which met the criteria and could be pooled were included. Meta-analysis of these trials for RF control produced a relative risk of 0.41 (95% CI: 0.23-0.70). There was statistical heterogeneity (I = 70.5%). Hence a random effects analysis was conducted. LIMITATIONS: Many studies were poor quality. Title and available abstracts of non-English studies were checked. There may be publication bias. This is the best available evidence in an area with imperfect information. DISCUSSION: It is expected acute RF cases would diminish by about 60% using a school or community clinic to treat streptococcal pharyngitis. This should be considered in high-risk populations. PMID- 19561422 TI - Enhanced immunogenicity of seasonal influenza vaccines in young children using MF59 adjuvant. AB - BACKGROUND: Children have high morbidity and hospitalization rates from seasonal influenza. Meta-analyses suggest that conventional inactivated influenza vaccines are of low efficacy in young children, making vaccines that induce greater and broader immune protection in this vulnerable population a medical priority. Adjuvanted influenza vaccines may offer a solution. SUBJECTS AND METHODS: Unprimed healthy children (6 to <36 months) were enrolled in an observer-blinded study and randomly assigned to receive 2 doses of MF59-adjuvanted vaccine (Sub/MF59, n = 130) or nonadjuvanted split vaccine (split, n = 139); subgroups of these (n = 43 and 46, respectively) received a booster dose 1 year later. Safety and clinical tolerability were assessed after each dose. Hemagglutination inhibition antibody titers were measured against influenza A and B strains included in the formulation of the vaccines and against mismatched strains. RESULTS: Clinical tolerability and safety were generally comparable between vaccine groups, though some transient, mild solicited reactions were more frequent in the Sub/MF59 group. Postvaccination hemagglutination inhibition antibody titers to all 3 vaccine strains were significantly higher with Sub/MF59 than with split vaccine (all comparisons P < 0.001) after each of the 3 vaccine doses. In addition, Sub/MF59 induced significantly higher cross-reactivity against A/H3N2 and A/H1N1 mismatched strains. CONCLUSION: MF59-adjuvanted influenza vaccine was well tolerated in healthy young children after each of 3 doses and induced greater, longer-lasting, and broader immune responses than a nonadjuvanted split vaccine. The enhanced immunogenicity of the adjuvanted vaccine was most evident in very young children and for the B vaccine strain. PMID- 19561423 TI - Outbreak of atypical pertussis detected by polymerase chain reaction in immunized preschool-aged children. AB - BACKGROUND: From October 2005 to March 2006, a laboratory-confirmed outbreak of pertussis occurred in preschool-aged children (1-4 years) in Toronto, Canada. A case-control study in children was done to identify the risk factors for being positive for Bordetella pertussis by polymerase chain reaction (PCR). METHODS: A case was defined as an individual 0 to 18 years of age who tested positive for B. pertussis by PCR in Toronto from October 2005 to March 2006. Each case was matched to 2 controls (negative for B. pertussis by PCR) by: (1) age, (2) geographic region of testing, and (3) date of testing. RESULTS: One hundred eighty-nine cases and 296 controls were enrolled. Only 42% of cases (vs. 25% controls, P < 0.0001) met a clinical definition of pertussis, 3 cases (4 controls) were hospitalized, and the secondary household attack rate was 8%. One third of cases were positive for another respiratory pathogen and >90% of cases and controls were up-to-date with pertussis immunization. Children attending school (matched odds ratio [ORm] = 5.2; 95% confidence intervals [CI]: 1.4-19.0), day care (ORm = 2.2; 95% CI: 1.2-4.0), visiting a doctor's office (ORm = 1.6; 95% CI: 1.0-2.5), or exposed to a household member (median age: 4-5 years) with a pertussis-like cough (ORm = 1.9; 95% CI: 1.0-3.6) were significantly more likely to have a positive PCR test for B. pertussis. CONCLUSIONS: The main risk factors for PCR positivity for B. pertussis were school or day care attendance. Atypical symptoms were likely moderated by high immunization rates or may have been caused by other respiratory pathogens. In some cases, a positive PCR result might simply have reflected transient nasopharyngeal carriage of B. pertussis. PMID- 19561424 TI - Prevalence and clinical characteristics of norovirus gastroenteritis among hospitalized children in Spain. AB - BACKGROUND: The importance of norovirus as a cause of gastroenteritis outbreaks is well documented, but the role of norovirus in sporadic acute severe gastroenteritis is not so well established. The aim of this study was to determine the prevalence and clinical characteristics of norovirus gastroenteritis among hospitalized children. METHODS: A prospective study was conducted in children less than 5 years old, admitted with acute gastroenteritis between January 2005 and January 2008 to the Pediatrics Department of the Universitary Hospital, Albacete, Spain. Demographic and clinical data were collected. A stool sample from each child was screened for enteropathogenic bacteria and tested by reverse transcription polymerase chain reaction for rotavirus, astrovirus, norovirus, and sapovirus and by immunochromatographic method for enteric adenoviruses. RESULTS: Norovirus was the second most frequent pathogen after rotavirus, being detected in 61 (17.3%) of the 352 children enrolled, in 29 of them (8.2%) as single agent. Mixed infections involving other viruses or bacteria were present in 52.4% of norovirus positive samples, a nosocomial source of infection was demonstrated in 17.2%. Norovirus infection was more prevalent in winter and affected mainly children less than 2 years of age. Vomiting was present in 68% and fever in 48.3% of cases, 3 children had nonfebrile seizures. Compared with rotavirus enteritis, norovirus infection was slightly less severe (in terms of severity score and need of intravenous rehydration) and fever was less frequent. CONCLUSIONS: Norovirus was a frequent cause of acute severe sporadic gastroenteritis in children representing the second etiologic agent after rotavirus. PMID- 19561426 TI - Rising rates of HIV infection among young US men who have sex with men. PMID- 19561425 TI - Incidence of persistent renal dysfunction in human immunodeficiency virus infected children: associations with the use of antiretrovirals, and other nephrotoxic medications and risk factors. AB - BACKGROUND: Survival of HIV-infected children continues to increase and the use of antiretrovirals (ARVs) is expanding; however there are few data regarding the incidence of renal dysfunction and associated risk factors among HIV-infected children and youth. METHODS: A total of 2102 children enrolled in Pediatric AIDS Clinical Trials Group Study 219/219C, were followed and assessed prospectively for >30 months. Occurrence of clinical events and laboratory abnormalities were recorded using standardized criteria and forms. Therapeutic decisions were made by clinicians at each site. Occurrence of persistent renal laboratory abnormalities was the main outcome measure. RESULTS: Four hundred forty-six (22%) enrollees exhibited at least one persistent renal laboratory abnormality. Elevated serum creatinine (Cr) was more common than persistent proteinuria (15% vs. 8%). The incidence of new renal laboratory abnormalities was 3.7 events per 100 person-years with rates increasing between 1993 and 2005. Older age (>or=6 years vs. <6 years), Hispanic ethnicity, and Black non-Hispanic race were associated with increased risk of renal dysfunction, but CDC clinical class and plasma HIV RNA levels were not. Subjects exposed to ARV regimens containing tenofovir and/or indinavir had approximately twice the risk of developing renal dysfunction compared with persons exposed to other ARVs. The risk of renal dysfunction was also elevated for other antivirals (hazard ratio = 5.4) and amphotericin B (hazard ratio = 28). CONCLUSIONS: Persistent renal function abnormalities occur frequently in HIV-infected children. Improved survival, Black race and Hispanic ethnicity, and exposure to tenofovir, indinavir, and other antimicrobial agents increase the risk for renal dysfunction. All HIV-infected children should be monitored closely for evidence of renal disease. PMID- 19561427 TI - Dengue fever and dengue hemorrhagic fever. PMID- 19561428 TI - Bacteriology of spontaneously draining acute otitis media in children before and after the introduction of pneumococcal vaccination. AB - Bacterial growth from spontaneously draining middle ear fluid was compared for periods before (1993-1998) and after (2001-2006) introduction of routine pneumococcal vaccination Staphylococcus aureus, including MRSA, was more common (P < .05) in the latter period. PMID- 19561429 TI - Incidence rate of nontuberculous mycobacterial disease in immunocompetent children: a prospective nationwide surveillance study in Germany. AB - An increasing incidence in disease caused by nontuberculous mycobacteria is being reported. We investigated the burden of disease in immunocompetent German children in a prospective nationwide study from April 2003 to September 2005. Ninety-seven percent of children presented with lymphadenitis; median age was 2.5 years. Using the capture-recapture method, we estimated a cumulative incidence rate of 3.1/100000 children. PMID- 19561430 TI - Pediatric and adolescent imported malaria in Cape Town. AB - We reviewed 42 cases of pediatric and adolescent imported malaria in Cape Town. Patients were predominantly new and returned immigrants from other African countries. Rapid diagnosis occurred in most cases. Eleven of 42 (26%) had severe malaria. Management issues included delay to and inappropriate treatment, inadequate monitoring for hypoglycemia, and under notification to health authorities. PMID- 19561432 TI - Congenital toxoplasmosis after a preconceptional or periconceptional maternal infection. AB - We report a case of asymptomatic congenital toxoplasmosis following a periconceptional infection of the mother. Fetal infection is very uncommon in such a situation, and mostly associated with fetal damage. We recommend that newborns undergo postnatal screening, even after maternal periconceptional infection, and receive specific therapy to reduce long-term sequelae. PMID- 19561431 TI - A phase I-II study of live attenuated varicella-zoster virus vaccine to boost immunity in human immunodeficiency virus-infected children with previous varicella. AB - Herpes zoster, may be severe and recurrent in HIV-infected children. We determined the safety and immunogenicity of live attenuated varicella-zoster virus (VZV) vaccine in 46 HIV-infected children who had experienced varicella. There were no serious adverse events. Two years after vaccination 82% of subjects remained VZV-antibody positive and 60% had VZV-specific cell-mediated immunity. No child developed herpes zoster. PMID- 19561433 TI - Parvovirus B19 infection associated with acute hepatitis in infant. PMID- 19561434 TI - Stability of 5% permethrin cream used for scabies treatment. PMID- 19561436 TI - Pancreatic pseudocyst rupture into the portal vein. AB - A patient with a pancreatic pseudocyst rupture into the portal vein with a resultant noninfectious systemic inflammatory response syndrome and subsequent portal vein thrombosis diagnosed by computed tomography and ultrasonography is reported. A review of the existing English literature on this rare complication is also provided. PMID- 19561437 TI - Validity and reliability of the Korean version of the AD8 informant interview (K AD8) in dementia. AB - The Alzheimer disease 8 (AD8) is a brief informant-based measure that distinguishes individuals with very mild dementia and mild cognitive impairment from those with normal cognition. The aim of this study was to establish the validity, reliability, and discriminative properties of the Korean version of the AD8 (K-AD8). Evaluation was made on 155 patient-informant dyads. The K-AD8 scores with the Clinical Dementia Rating (CDR) and performance on neuropsychologic tests were compared. Internal consistency of the K-AD8 was significant (Cronbach alpha=0.88). The K-AD8 had strong correlation with CDR (Spearman rho=0.76). Concurrent validity was strong with the K-AD8 scores correlating with CDR domains and performance on neuropsychologic tests. The K-AD8 was found to have excellent test-retest reliability (weighted kappa=0.81) and good interrater reliability (intraclass correlation coefficient=0.89). The area under the receiver operator characteristic curve was 0.88, suggesting good discrimination between nondemented individuals and those with cognitive impairment. To distinguish CDR 0.5 from CDR 0, in particular, the K-AD8 (cutoff score of 2) had a sensitivity of 68% and a specificity of 90%. In conclusion, the K-AD8 is a sensitive screening tool in detecting very early dementia, indicating that the AD8 could work very well in a variety of cultural entities. PMID- 19561438 TI - Predictors of physician referral for patient recruitment to Alzheimer disease clinical trials. AB - BACKGROUND: Inadequate recruitment into Alzheimer disease clinical trials is an important threat to the validity and generalizability of the studies. The majority of dementia patients are first evaluated by community-based physicians; however, physician perceptions of clinical research are largely unknown. METHODS: A survey was distributed to 3123 physicians in 3 states; 370 were returned. Survey items assessed attitudes, perceived benefits of and barriers to referral to clinical research, and physicians use of the internet for medical information. RESULTS: The mean age of the respondents was 50.6+/-10.8 years; 70% were male, 78% white, 61% were primary care providers; 63% used the internet > or =3 times/week. No demographic or medical specialty differences existed between those who were likely (n=193) and unlikely (n=162) to refer patients to clinical trials. Differences were discovered in perceived benefits reported by physicians who were more likely to refer, whereas differences in perceived barriers existed in primary care compared with specialists. Referral to clinical trials is predicted by close proximity to a research center [odds ratio (OR): 4.0; 95% confidence interval (CI), 1.1-15.6] and availability of internet information regarding diagnostic evaluation (OR: 2.3; 95% CI, 1.1-4.7). Primary barriers included concerns about exposure of patients to uncomfortable procedures (OR: 4.7; 95% CI, 1.2-18.7) and lack of time to discuss research participation (OR: 6.8; 95% CI, 1.4-32.3). CONCLUSIONS: Proximity to a research center and availability of diagnostic clinical tools are strong predictors of clinical trial referral. Concern over risks to patients and lack of time are strong barriers. These results suggest that dementia outreach education targeted to physicians should emphasize the importance of clinical trials with a focus on discussing research participation in a time-efficient manner and increasing awareness of risk reduction and the safety of research protocols. Providing easy access to up to-date, user-friendly educational materials on dementia diagnosis and research via the internet are likely to improve referrals of patients to Alzheimer disease clinical trials from community physicians. PMID- 19561439 TI - A battery of tests for assessing cognitive function in older Latino persons. AB - With the proportion of older Latinos in the United States rapidly growing, dementia is expected to be an increasing public health problem in this segment of the population. Yet relatively few cognitive test batteries have been developed for evaluating older Spanish-speaking persons. We selected a battery of cognitive tests used in cognitive aging studies of English speakers, adapted it for Spanish speakers, and administered it to 66 older Latinos (mean age=71.1, SD=8.1). The results of a factor analysis supported grouping the tests into the same 5 functional domains identified for English speakers. Composite measures of performance in each domain were positively related to education and, with some exceptions, inversely related to age. The results suggest that this battery may be useful in epidemiologic research on cognition in older Latinos. PMID- 19561440 TI - Rapidly progressive dementia: causes found in a Greek tertiary referral center in Athens. AB - Dementia is generally considered as rapidly progressive [rapidly progressive dementia (RPD)], in cases with overt cognitive impairment, established within months. Data about the relative frequency of underlying diseases in cases of RPD are few and extremely variable, depending on the clinical setting. We examined the relative frequency of the underlying causes of RPD, in a university tertiary referral center, in Athens. A series of consecutive patients presenting with RPD in a 3-year period was included. All patients received a comprehensive clinical, imaging, and laboratory evaluation. Of a total of 279 patients hospitalized for dementia diagnosis, 68 patients had RPD (37 males and 31 females). Mean age at onset +/-SD was 65.5+/-10.0. The most common cause of RPD was secondary dementias, accounting for 18 cases (26.5%). Alzheimer disease and frontotemporal dementia were almost equally represented, accounting for 12 (17.6%) and 11 (16.2%) cases, respectively. Vascular dementia, Creutzfeldt-Jakob disease, and various neurodegenerative diseases accounted for 9 cases each (13.2%). In a tertiary referral center, secondary dementias represented the most frequent cause of cases presenting with RPD. As a substantial number of these cases are potentially treatable, our finding reconfirms and underscores the importance of an exhaustive evaluation in any case presenting with RPD. PMID- 19561441 TI - Polypharmacy and potentially inappropriate medication use among community dwelling elders with dementia. AB - This cross-sectional study examines the association between total prescription medication use and potentially inappropriate medication use (PIRx) among community-dwelling elderly patients with and without dementia. Data (September 2005 to September 2007) were from the National Institute on Aging-funded National Alzheimer's Coordinating Center Uniform Data Set. The study analyzed the Uniform Data Set initial visits of 4518 community-dwelling subjects aged 65 years and above with and without dementia (2665 and 1853, respectively). PIRx was defined using a partial list of the 2003 Beers criteria. Generalized linear mixed models were applied to estimate the association between PIRx and polypharmacy. In both groups (with and without dementia), subjects who received PIRx on average took more medications than those taking no PIRx. As the total number of medications used increased, the odds of having PIRx also increased, controlling for dementia diagnosis and other subject characteristics. Our key findings were consistent after considering 2 definitions of PIRx (with or without oral estrogens) and accounting for missing data. In summary, the total number of medications used is associated with PIRx among Alzheimer's Disease Centers community-dwelling elderly patients with and without dementia, with polypharmacy increasing the risk of PIRx. Ensuring appropriate medication use in this population is clinically important because of the significant risks for institutionalization. PMID- 19561442 TI - Increasing hippocampal atrophy and cerebrovascular damage is differently associated with functional cortical coupling in MCI patients. AB - The aim of this study was to assess the changes of intrahemispheric and interhemispheric linear spectral electroencephalography (EEG) coherence associated with increasing hippocampal atrophy (HA) and white matter hyperintensities (WMHs) in patients with mild cognitive impairment (MCI). Eighty five MCI patients underwent clinical and neuropsychologic assessment, EEG recordings, and magnetic resonance imaging. Intrahemispheric (fronto-temporal, temporo-parietal, and fronto-parietal) and interhemispheric (frontal, temporal, and parietal) linear EEG coherence was computed. MCI patients were categorized into classes of increasing HA and WMHs on the basis of tertile distribution of volume loss (high, mid, and low). Neuropsychologic tests were also gathered and compared in MCI subgroups. As expected, MCI patients with high WMHs had poorer performance on visuospatial and cognitive flexibility, whereas those with high HA failed on memory tests. Significant differences of EEG functional coupling were present in the fronto-temporal network in patients with high WMHs and high HA, but without a different pattern. In high WMHs the EEG coherence in low frequencies was increased (with the exception of alpha1 band), whereas coherence in the fast frequencies was decreased proportional to increasing damage. In high HA a change of coherence was present in the delta and alpha2 frequency bands that was not proportional to HA, fast frequencies being unaffected. Our results show a lateralization (right hemisphere for cerebrovascular disease and left hemisphere for hippocampal atrophy) of the pathologic modifications of functional coupling. PMID- 19561444 TI - Rates of cognitive change in Alzheimer disease: observations across a decade of placebo-controlled clinical trials with donepezil. AB - Treatment success in Alzheimer disease (AD) trials is generally based on benefits over placebo-treated controls. Consequently, variation in rates of decline among placebo-treated patients could impact outcomes from AD trials. In the present analyses, individual patient data [baseline Mini-Mental State Examination (MMSE): 10 to 26] were pooled from randomized, placebo-controlled studies of donepezil for AD conducted during the 1990s, and grouped by initiation year-group 1: 1990 to 1994; group 2: 1996 to 1999. Changes in MMSE and Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog) were compared between groups 1 and 2 for placebo, and then between donepezil and placebo. Data were available from 3403 patients in 13 trials. Group 2 (post-1995) included patients with lower baseline MMSE scores, older patients, fewer males, more comorbidity, and more concomitant medications. MMSE decline by week 24 was significantly greater among group 1 (pre-1995) placebo patients versus group 2; a similar trend was observed with the ADAS-cog. Nevertheless, donepezil-mediated treatment effects were consistent over the decade of enrollment. These analyses suggest that patients are showing slower rates of cognitive decline in more recent trials compared with older trials, although having more comorbidities. This finding may have important potential implications for future clinical trial design. PMID- 19561443 TI - Gelsolin levels are increased in the brain as a function of age during normal development in children that are further increased in Down syndrome. AB - Neuronal dysfunctions in several neurodegenerative diseases such as Down syndrome (DS) have been linked to oxidative stress. In this study, we observed that lipid peroxidation, a marker of oxidative stress, is significantly increased in the frontal cortex of brains of individuals with DS as compared with control subjects. We report here that gelsolin levels are increased in the frontal cortex of individuals with DS as compared with controls during early developmental ages (5 to 13 y). Interestingly, the levels of gelsolin in the frontal cortex were increased as a function of age in both DS and control subjects. Because cytoplasmic gelsolin has 5 free thiol groups (cysteine), and its levels are increased in response to oxidative stress, we propose that gelsolin may serve as an antioxidant protein. PMID- 19561445 TI - Testicular germ cell tumors with sarcomatous components: an analysis of 33 cases. AB - The development of sarcomatous component (SC) in testicular germ cell tumor (GCT) is an uncommon phenomenon. We searched our surgical pathology files from 1985 to 2007 and identified 33 cases of testicular GCTs with SC. The average age of patients was 31 years. All patients underwent radical orchiectomy, which demonstrated a GCT in all patients except for 3 patients who had received neoadjuvant chemotherapy. All testicular GCTs contained a teratomatous component. The GCTs were pure teratomas in 3 cases, and were mixed GCTs in the other cases. The SC was observed in primary testicular tumor (n=19), in metastasis (n=11), or in both primary testicular tumor and metastasis (n=3). The average percentage of the SC in the primary testicular GCT was 32% (range: 5% to 99%). The most common histologic type of SC was rhabdomyosarcoma (n=24), followed by high-grade unclassified sarcoma (n=5), rhabdomyosarcoma admixed with high-grade unclassified sarcoma (n=2), angiosarcoma (n=1), and low-grade myxoid sarcoma (n=1). Clinical follow-up information was available for 27 patients. Of the 13 patients whose SC was limited to the testicular GCT, 2 patients died of GCT not otherwise specified at 37 and 68 months, respectively; and 11 patients were free of disease at a mean of 46 months. Of the 14 patients with a SC in the metastasis, 7 patients died of GCT not otherwise specified at a mean of 95 months, and 7 patients were free of disease at a mean of 104 months. These results suggest that patients with a SC confined to the primary testicular GCT may not have a higher risk of mortality than those at a comparable stage without a SC. However, patients with a SC in the metastasis have an increased risk of mortality. PMID- 19561447 TI - IgG4-related sclerosing pachymeningitis: a previously unrecognized form of central nervous system involvement in IgG4-related sclerosing disease. AB - IgG4-related sclerosing disease is a distinctive mass-forming lesion with frequent systemic involvement, most frequently the pancreas, salivary glands, and lacrimal glands. This report describes a case manifesting with a previously unrecognized form of central nervous system involvement. The 37-year-old man presented with signs and symptoms of spinal cord compression at the thoracic level 9. Magnetic resonance imaging revealed an elongated dural mass extending from the fifth to tenth thoracic vertebra. Laminectomy and excision of the mass revealed dura expanded by a dense lymphoplasmacytic infiltrate accompanied by stromal fibrosis and phlebitis. IgG4+ plasma cells were increased and the proportion of IgG4+/IgG+ plasma cells was 85%. The patient also had a 1-year history of bilateral submandibular swelling due to chronic sialadenitis. Thus, IgG4-related sclerosing pachymeningitis represents a new member of the IgG4 related sclerosing disease family affecting the central nervous system. It seems that at least a proportion of cases described in the literature as idiopathic hypertrophic pachymeningitis belong to this disease, especially as some patients have other clinical manifestations compatible with IgG4-related sclerosing disease, such as cholangitis and orbital pseudotumor. PMID- 19561446 TI - NUT midline carcinoma of the parotid gland with mesenchymal differentiation. AB - Nuclear protein in testis midline carcinomas (NMC) are highly aggressive carcinomas typically arising in midline structures in young individuals. These carcinomas are characterized by the presence of a chromosomal rearrangement of nuclear protein in testis the (NUT) gene on chromosome 15 (15q14), resulting from a chromosomal translocation most commonly involving the BRD4 gene on chromosome 19p13. Rarely, in about 1/3 of cases, other translocation partners are involved (termed NUT-variants). Most cases have involved midline structures and with few exceptions were located in the upper aerodigestive tract and the mediastinum. Except for a single case, all reported NMC have been fatal, proving resistant to multimodality treatment. We report an exceptional case of a NMC presenting outside of midline structures in the parotid gland and showing mesenchymal chondroid differentiation in a 15-year-old male. The presence of the t(15;19) chromosomal translocation in the chondroid component was confirmed by fluorescence in situ hybridization analysis and immunohistochemical staining, indicating mesenchymal transdifferentation of the tumor. The findings demonstrate the first case of NMC arising within salivary gland, and the first example of mesenchymal differentiation in this group of tumors. PMID- 19561448 TI - Prevalence and origin of amyloid in kidney biopsies. AB - We aimed to reassess renal amyloidosis in kidney biopsies with a focus on possibly misclassified or unclassified cases and changes in the prevalence of different amyloid types. Two hundred thirty-three kidney biopsies obtained from 231 patients diagnosed with amyloid during the period from 1990 to 2007 years were included in this retrospective study. Amyloid was identified by Congo red staining and polarization microscopy. Immunohistochemical classification was made with antibodies directed against AA amyloid, apolipoprotein A1, fibrinogen, lysozyme, lambda-light chain, kappa-light chain, beta2-microglobulin, transthyretin, and amyloid P-component. Amyloid was present in each biopsy as vascular, tubulo-interstitial and/or glomerular deposits. Immunoglobulin light chain-derived (AL) amyloidosis was most prevalent and diagnosed in 123 (53.2%) patients. It was categorized into AL amyloid of lambda-light chain (ALlambda) [105 (85.4%) patients] and kappa-light chain origin (ALkappa) [10 (8.1%)]. The amyloid deposits of 8 (6.5%) patients were not clearly distinguishable into ALlambda amyloid or ALkappa amyloid and categorized as AL amyloid, not otherwise specified. Reactive systemic amyloid A (AA) amyloidosis was the second most common type and was found in 93 patients (40.3%). Overall 7 patients were found to suffer from fibrinogen A alpha-chain-[amyloid of fibrinogen (AFib); 4 (1.7%) patients], transthyretin-[amyloid of transthyretin (ATTR); 2 (0.9%)], or apolipoprotein A1-derived (AApoAI) amyloidosis [1 (0.4%)]. In 8 patients (3.4%) the amyloid deposits remained unclassifiable. After additional immunostaining and further clinical information the diagnoses of 12 patients (5.1%) were modified (2 ALlambda amyloid, 4 ALkappa amyloid, 1 amyloid unclassified, 3 mixed-type amyloidosis, AA+ATTR, ALlambda+ATTR, and ALkappa+ATTR, 1 AFib, and 1 AApoAI). Although the histologic and immunohistochemical reevaluation confirmed the classifications in 221 (95.7%) patients. Renal amyloidosis is most commonly of ALlambda-origin, followed by AA amyloidosis. AFib amyloidosis was found to be the most prevalent type of hereditary renal amyloidosis, illustrating the necessity of a thorough classification of the amyloid proteins. PMID- 19561449 TI - NK-cell lineage predicts poor survival in primary intestinal NK-cell and T-cell lymphomas. AB - Most primary intestinal natural killer (NK)-cell and T-cell lymphomas (PINKTL) in the Northern Europe are enteropathy-associated T-cell lymphomas, a complication of celiac disease, which is rare in the East. Primary intestinal NK-cell lymphoma is extremely rare and is poorly characterized. We investigated 30 cases of PINKTL from Taiwan with male: female at 2:1, median age at 55.5, 80% with jejunal/ileal involvement, 77% with perforation, 27% with multicentric tumors, and 67% at stage IE. All 7 cases tested for serum IgA anti-tissue transglutaminase were negative. Only 3 (10%) tumors showed enteropathy. Six (20%) were NK-cell lymphoma and 24 (80%) were T-cell lymphoma. The tumor cells in 21/30 (70%) cases were small to medium sized, which correlated with the coexpression of both CD8 and CD56. All tumors expressed at least 1 cytotoxic marker. All 6 NK-cell lymphomas were negative for betaF1, diffusely positive for Epstein-Barr virus-encoded mRNA (EBER), and polyclonal for T-cell receptor gene rearrangement. Five (22%) of the 24 T-cell tumors expressed betaF1, 8 (35%) of the 23 tumors were positive for EBER, and 20 (95%) of the 21 tumors were clonal for T-cell receptor. The overall 1-year survival was 36%. Univariate regression analysis showed that NK-cell lineage, multicentricity, and perforation were associated with poor prognosis. NK cell lineage (P=0.037) was a poor prognostic factor by multivariate Cox proportional hazard regression analysis. PINKTL in Taiwan is predominantly not enteropathic with a high frequency of perforation, small to medium tumor cell size and cytotoxic phenotype. Primary intestinal NK-cell lymphoma carries a very poor prognosis, and is probably a distinct entity. PMID- 19561451 TI - Rifampin-associated pseudomembranous colitis. AB - Pseudomembranous colitis (PMC) is known to develop after antibiotic treatment, but is rarely associated with antituberculosis (anti-TB) agents. We report a 28 year-old woman without underlying diseases developing PMC after 126 days of anti TB treatment. Severe diarrhea and abdominal cramping pain were experienced. Colonoscopic biopsy proved the diagnosis of PMC. Her symptoms improved after discontinuing the anti-TB agents but recurred shortly after challenging with rifampin and isoniazid. Metronidazole administration and replacement of rifampin with levofloxacin successfully cured the PMC. Our report supports the notion that rifampin can induce PMC. PMID- 19561450 TI - Fluorescence in situ hybridization (FISH) as an ancillary diagnostic tool in the diagnosis of melanoma. AB - Although the clinical and pathologic diagnosis of some melanomas is clear-cut, there are many histopathologic simulators of melanoma that pose problems. Over diagnosis of melanoma can lead to inappropriate therapy and psychologic burdens, whereas under-diagnosis can lead to inadequate treatment of a deadly cancer. We used existing data on DNA copy number alterations in melanoma to assemble panels of fluorescence in situ hybridization (FISH) probes suitable for the analysis of paraffin-embedded tissue. Using FISH data from a training set of 301 tumors, we established a discriminatory algorithm and validated it on an independent set of 169 unequivocal nevi and melanomas as well as 27 cases with ambiguous pathology, for which we had long-term follow-up data. An algorithm-using signal counts from a combination of 4 probes targeting chromosome 6p25, 6 centromere, 6q23, and 11q13 provided the highest diagnostic discrimination. This algorithm correctly classified melanoma with 86.7% sensitivity and 95.4% specificity in the validation cohort. The test also correctly identified as melanoma all 6 of 6 cases with ambiguous pathology that later metastasized. There was a significant difference in the metastasis free survival between test-positive and negative cases with ambiguous pathology (P=0.003). Sufficient chromosomal alterations are present in melanoma that a limited panel of FISH probes can distinguish most melanomas from most nevi, providing useful diagnostic information in cases that cannot be classified reliably by current methods. As a diagnostic aid to traditional histologic evaluation, this assay can have significant clinical impact and improve classification of melanocytic neoplasms with conflicting morphologic criteria. PMID- 19561453 TI - Mitral valve prolapse with a late-systolic regurgitant murmur may be associated with significant hemodynamic consequences. AB - The late-systolic murmur of mitral regurgitation (MR) in degenerative mitral valve disease is widely believed to represent regurgitation of a degree that is not associated with hemodynamic significance. However, the extent of left ventricular (LV) remodeling associated with the late-systolic murmur has not been systematically assessed. Accordingly, we studied 82 patients sent for evaluation of at least moderate isolated MR by echocardiography/Doppler examination. All patients had a physical examination and cardiac magnetic resonance imaging to measure LV volumes by summation of serial short-axis slices. Forty-five patients had a pan-systolic murmur and 37 had a late-systolic murmur on auscultation that was verified by timing of onset of regurgitant turbulence by cine magnetic resonance imaging. Systolic blood pressures (124 +/- 3 versus 124 +/- 3 mm Hg) and LV ejection fraction (61 +/- 1% versus 61 +/- 1%) did not differ significantly between pan-systolic and late-systolic murmur groups. Although LV end-diastolic volume index was greater in the pan-systolic versus late-systolic murmur (108 +/- 4 versus 95 +/- 4 mL/m2, P = 0.007), both groups were significantly greater than normals (68 +/- 2 mL/m2, P < 0.0001). However, LV end systolic volume index (42 +/- 2 versus 38 +/- 2 mL/m2) and LV end-systolic dimension (38 +/- 1 versus 37 +/- 1 mm), critical markers of adverse LV remodeling in isolated MR, did not differ significantly between pan-systolic and late-systolic murmur groups. In conclusion, the late systolic isolated MR murmur may be associated with significant adverse LV remodeling, and should not be considered evidence of hemodynamically unimportant MR. PMID- 19561452 TI - Increased frequency of prediabetes in patients with irritable bowel syndrome. AB - The aim of this study was to compare the occurrence of prediabetes [impaired fasting glucose and/or impaired glucose tolerance are considered to be precursors to type 2 diabetes mellitus (DM)] in irritable bowel syndrome (IBS) cases and matched controls. Ninety-two patients with IBS and 104 healthy matched controls were included in this study. Type 2 DM was considered an exclusion criterion in both groups. Fasting blood glucose, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and triglyceride levels were examined; after 1 night of fasting, an oral glucose tolerance test with 75 g glucose was administered, and the blood glucose levels after 2 hours were examined. Although there were no significant differences in the triglyceride levels, significant differences were found for total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels (P < 0.001, 0.001, and <0.001, respectively). These measures were found to be elevated in the IBS group compared with the control group. The frequency of prediabetes, which is regarded as the first stage of type 2 DM, was also found to be significantly higher in the IBS group (P < 0.001). After adjusting for potential confounders, such as age, lipid levels, and anthropometric measures in the analysis of covariance models, prediabetes was significantly more frequent in the IBS group than in the control group (P < 0.001). Thus, given the higher prediabetes occurrence in IBS, IBS may indirectly indicate a higher risk of DM. Further investigations will be necessary to fully elucidate the mechanisms behind these observations. PMID- 19561454 TI - Anesthesia and pharmacogenetics. PMID- 19561455 TI - Modification of cysteinyl leukotriene receptors expression in capsular contracture: follow-up study and definitive results. AB - Periprosthetic capsular contracture represents a specific iatrogenic phenomenon with different side effects. Recently, interesting data have disclosed a potential role of leukotrienes as important mediators of inflammation in the reactivation process of capsular contracture. Some preliminary studies have assessed the efficacy of leukotriene antagonists in the prevention and treatment of capsular contracture. These clinical data are still lacking of a potential biomolecular basis. The aim of our present study was to evaluate the expression of the protein receptor cysteinyl leukotriene receptors (CysLTR). We included 50 patients with severe capsular contracture (Baker III-IV) and a control group consisting of healthy patients who underwent an implant replacement. In both groups, we performed the protein extraction and semiquantitative analysis for the determination of protein concentration on myofibroblasts and macrophages. Western Blot analysis of protein levels shows a significant increase in the expression of CysLTR in patients with capsular contracture. Our final results show that the increase in the levels of mRNA coding for CysLTR actually translates into an effective increase in protein levels of these mRNA transcripts. These findings could at least partially provide a biomolecular basis that justifies the use of specific antileukotriene drugs in the treatment of this disease. PMID- 19561456 TI - The obesity paradox: body mass index and outcomes in patients undergoing nonbariatric general surgery. AB - OBJECTIVE: We sought to examine the effect of body mass index (BMI) on 30-day morbidity and mortality in a large cohort of patients undergoing nonbariatric general surgery. SUMMARY BACKGROUND DATA: Obesity has long been considered a risk factor for poor outcomes from a variety of surgical procedures, yet recent studies of critically and chronically ill patients suggest that overweight and obese patients may paradoxically have better outcomes than "normal" weight patients. METHODS: A prospective, multi-institutional, risk-adjusted cohort study of 118,707 patients undergoing nonbariatric general surgery who were included in the National Surgical Quality Improvement Program Participant Use database in 2005 and 2006 was performed. Outcomes and risk variables were compared across NIH defined BMI class using analysis of variance, Bonferroni multiple comparisons of means tests, and multivariable logistic regression. RESULTS: After adjusting for all significant perioperative risk factors, the risk of death according to BMI exhibited a reverse J-shaped relationship, with the highest rates in the underweight and morbidly obese extremes and the lowest rates in the overweight and moderately obese. Overweight (odds ratio, 0.85; 95% CI, 0.75-0.99) and moderately obese (odds ratio, 0.73; 95% CI, 0.57-0.94) patients had a significantly lower risk of death than normal weight patients. There was a progressive increase in the likelihood of a complication with increasing BMI class, almost entirely due to increasing rates of wound infection. CONCLUSIONS: Overweight and moderately obese patients undergoing nonbariatric general surgery have paradoxically "lower" crude and adjusted risks of mortality compared with patients at a "normal" weight. This finding is in contrast to observations from the general population, confirming the existence of an "obesity paradox" in this patient population. PMID- 19561457 TI - Evolution of the surgeon-volume, patient-outcome relationship. AB - OBJECTIVE: Higher surgeon volume is associated with improved patient outcomes. This finding has prompted recommendations for increasing specialization and referrals to high-volume surgeons, yet their implementation in clinical practice has not been measured. METHODS: We performed cross-sectional analyses using 1999 and 2005 discharge information from the Health Care Utilization Project National Inpatient Sample to measure whether the number of procedures performed by high volume surgeons increased over time. Procedures included those demonstrated to have strong surgeon volume-outcome associations in the literature. International Classification of Diseases, Ninth Revision codes were employed for colorectal procedures, esophagectomy, gastrectomy, pancreatectomy, thyroidectomy, coronary artery bypass graft surgery, and carotid endarterectomy. Bivariate analyses and hierarchical generalized linear models were employed to measure association between surgeon volume and length of stay (LOS) and mortality or complications. RESULTS: There was a significant increase in the proportion of procedures performed by high-volume surgeons over time, with the most dramatic increases seen for gastrectomy (54%), pancreatectomy (31%), and thyroidectomy (23%). Having a procedure performed by a high-volume surgeon was associated with patient race and insurance status. Overall, unadjusted mortality and LOS were significantly lower for high-volume surgeons compared with low-volume surgeons in 1999 and 2005. In multivariable hierarchical generalized linear models, only differences in LOS by surgeon volume remained significant in both years. CONCLUSIONS: The proportion of procedures performed by high-volume surgeons increased over a 6 year period, as evidence mounted in support of a surgeon volume-outcome association. Efforts are still needed to improve access among underserved subsets of the population and eliminate apparent disparities based on patient race and insurance status. PMID- 19561458 TI - Negative impact of neutrophil-lymphocyte ratio on outcome after liver transplantation for hepatocellular carcinoma. AB - BACKGROUND: The Milan criteria have been adopted by United Network for Organ Sharing (UNOS) to preoperatively assess outcome in patients with hepatocellular carcinoma (HCC) who receive orthotopic liver transplantation (OLT). These criteria rely solely on radiographic appearances of the tumor, providing no measure of tumor biology. Recurrence rates, therefore, remain around 20% for patients within the criteria. The neutrophil-lymphocyte ratio (NLR) is an indicator of inflammatory status previously established as a prognostic indicator in colorectal liver metastases. We aimed to determine whether NLR predicts outcome in patients undergoing OLT for HCC. DESIGN: Analysis of patients undergoing OLT for HCC between 2001 and 2007 at our institution. A NLR > or =5 was considered to be elevated. RESULTS: : A total of 150 patients were identified, with 13 patients having an elevated NLR. Of these, 62% developed recurrence compared with 14% with normal NLR (P < 0.0001). The disease-free survival for patients with high NLR was significantly worse than that for patients with normal NLR (1-, 3-, and 5-year survivals of 38%, 25%, and 25% vs. 92%, 85%, and 75%, P < 0.0001). Patients with high NLR also had poorer overall survival (5-year survival, 28% vs. 64%, P = 0.001). Patients within Milan with an elevated NLR had significantly poorer disease-free survival than those with normal NLR within Milan (5-year survival, 30% vs. 81%, P < 0.0001). On univariate analysis, 9 factors including an NLR > or =5 were significant predictors of poor disease-free survival. However, only a raised NLR remained significant on multivariate analysis (P = 0.005, HR: 19.98). CONCLUSION: Elevated NLR significantly increases the risk for tumor recurrence and recipient death. Preoperative NLR measurement may provide a simple method of identifying patients with poorer prognosis and act as an adjunct to Milan in determining, which patients benefit most from OLT. PMID- 19561459 TI - Saline-linked surface radiofrequency ablation: a safe and effective method of surface ablation of hepatic metastatic colorectal cancer. AB - OBJECTIVE: To determine the safety and efficacy of saline-linked surface radiofrequency ablation (SLSRFA) in a clinical setting. SUMMARY BACKGROUND DATA: We have previously identified safe and effective parameters for use of SLSRFA in a porcine model. METHODS: An initial study was conducted to determine if parameters defined in the porcine model were safe and effective in human livers. In 16 patients undergoing liver resection, normal areas of liver were treated with SLSRFA using various power/diameter combinations (10 W/1 cm; 15 W/2 cm; 45 W/4 cm) for 9 minutes with and without inflow occlusion. In a second study, superficial hepatic colorectal cancer (CRC) metastases were treated at 45 W/4 cm for 9 minutes without inflow occlusion in 11 patients. Ablation depth was measured and samples were examined for cell viability by nicotine adenine dinucleotide stain. This study was registered in the ClinicalTrials.gov database and has the following ID number, NCT00869843. RESULTS: Ablation depth in normal liver varied from 3 to 20 mm. Depth was significantly dependent on power, lesion size, and inflow occlusion. Nicotine adenine dinucleotide stains showed total cell necrosis to the full depth of ablation. In the second study, large hepatic CRC metastases showed total cell necrosis to a mean depth of 12 mm. Two tumors less than 7 mm in depth showed complete necrosis. Metastases were more susceptible to SLSRFA than normal liver. CONCLUSION: SLSRFA completely and safely ablates normal liver to a depth of at least 4 mm at 45 W/4 cm treatment parameters. Remarkably, it is even more effective in ablating metastatic CRC. SLSRFA is an effective tool for extending resection margins and for ablating superficial small tumors or superficial parts of large tumors. PMID- 19561460 TI - Early endoscopic retrograde cholangiopancreatography in predicted severe acute biliary pancreatitis: a prospective multicenter study. AB - SUMMARY BACKGROUND DATA: The role of early endoscopic retrograde cholangiopancreatography (ERCP) in acute biliary pancreatitis (ABP) remains controversial. Previous studies have included only a relatively small number of patients with predicted severe ABP. We investigated the clinical effects of early ERCP in these patients. METHODS: We performed a prospective, observational multicenter study in 8 university medical centers and 7 major teaching hospitals. One hundred fifty-three patients with predicted severe ABP without cholangitis enrolled in a randomized multicenter trial on probiotic prophylaxis in acute pancreatitis were prospectively followed. Conservative treatment or ERCP within 72 hours after symptom onset (at discretion of the treating physician) were compared for complications and mortality. Patients without and with cholestasis (bilirubin: >2.3 mg/dL [40 mumol/L] and/or dilated common bile duct) were analyzed separately. RESULTS: Of the 153 patients, 81 (53%) underwent ERCP and 72 (47%) conservative treatment. Groups were highly comparable at baseline. Seventy eight patients (51%) had cholestasis. In patients with cholestasis, ERCP (52/78 patients: 67%), as compared with conservative treatment, was associated with fewer complications (25% vs. 54%, P = 0.020, multivariate adjusted odds ratio [OR]: 0.35, 95% confidence interval [CI]: 0.13-0.99, P= 0.049). This included fewer patients with >30% pancreatic necrosis (8% vs. 31%, P = 0.010). Mortality was nonsignificantly lower after ERCP (6% vs. 15%, P = 0.213, multivariate adjusted OR: 0.44, 95% CI: 0.08-2.28, P = 0.330). In patients without cholestasis, ERCP (29/75 patients: 39%) was not associated with reduced complications (45% vs. 41%, P = 0.814, multivariate adjusted OR: 1.36; 95% CI: 0.49-3.76; P = 0.554) or mortality (14% vs. 17%, P = 0.754, multivariate adjusted OR: 0.78; 95% CI: 0.19-3.12, P = 0.734). CONCLUSIONS: Early ERCP is associated with fewer complications in predicted severe ABP if cholestasis is present. PMID- 19561461 TI - Stem cells in sepsis. AB - OBJECTIVE: To review the characteristics of stem cells that may qualify them to be useful as therapeutic agents in sepsis. SUMMARY BACKGROUND DATA: Sepsis is a devastating syndrome and is the leading cause of death among critically ill surgical patients in the United States. Despite decades of research and numerous clinical trials, little progress has been made in the development of new treatments and mortality rates are much the same as they have been for the last 20 to 30 years. As such, sepsis remains a formidable adversary for surgeons and their patients and new therapeutic modalities must continue to be explored. METHODS: Recent literature regarding sepsis and the use of stem cells in the treatment of various inflammatory conditions including sepsis was reviewed. Our experience with the use of stem cells in our own laboratory was included. RESULTS: Stem cells have recently emerged as a promising therapy for a variety of commonly encountered surgical pathologies including cardiovascular disease, neurodegenerative disease, peripheral vascular disease, renal disease, and several others. Their beneficial effects are owed chiefly to their abilities to home to sites of injury and inflammation, to attenuate the inflammatory response, and to accelerate tissue healing and neoangiogenesis in the face of noxious stimuli. CONCLUSIONS: Experimental evidence indicates that stem cells are immunologically responsive cells that home to sites of inflammation and tissue injury. Stem cells also secrete growth factors in response to lipopolysaccharide and tumor necrosis factor that may limit apoptosis and organ injury. Stem cells represent an endogenous therapeutic strategy that may be enhanced for maximum clinical benefit. PMID- 19561462 TI - Normothermic liver preservation: is it time to come in from the cold? PMID- 19561463 TI - Normothermic perfusion: a new paradigm for organ preservation. AB - OBJECTIVE: Transplantation of organs retrieved after cardiac arrest could increase the donor organ supply. However, the combination of warm ischemia and cold preservation is highly detrimental to the reperfused organ. Our objective was to maintain physiological temperature and organ function during preservation and thereby alleviate this injury and allow successful transplantation. BACKGROUND DATA: We have developed a liver perfusion device that maintains physiological temperature with provision of oxygen and nutrition. Reperfusion experiments suggested that this allows recovery of ischemic damage. METHODS: In a pig liver transplant model, we compared the outcome following either conventional cold preservation or warm preservation. Preservation periods of 5 and 20 hours and durations of warm ischemia of 40 and 60 minutes were tested. RESULTS: After 20 hours preservation without warm ischemia, post-transplant survival was improved (27%-86%, P = 0.026), with corresponding differences in transaminase levels and histological analysis. With the addition of 40 minutes warm ischemia, the differences were even more marked (cold vs. warm groups 0% vs. 83%, P = 0.001). However, with 60 minutes warm ischemia and 20 hours preservation, there were no survivors. Analysis of hemodynamic and liver function data during perfusion showed several factors to be predictive of posttransplant survival, including bile production, base excess, portal vein flow, and hepatocellular enzymes. CONCLUSIONS: Organ preservation by warm perfusion, maintaining physiological pressure and flow parameters, has enabled prolonged preservation and successful transplantation of both normal livers and those with substantial ischemic damage. This technique has the potential to address the shortage of organs for transplantation. PMID- 19561464 TI - Anterior perineal plane for ultra-low anterior resection of the rectum (The APPEAR Technique): a prospective clinical trial of a new procedure. PMID- 19561466 TI - Reporting randomized trials. PMID- 19561467 TI - Impact of laparoscopic colorectal surgery on oncological outcomes. PMID- 19561469 TI - Laparoscopic resection for diverticular disease: follow-up of 500 consecutive patients. PMID- 19561471 TI - Evaluating the surgery literature: can standardizing peer-review today predict manuscript impact tomorrow? AB - OBJECTIVE: Evidence-based surgery is predicated on the quality of published literature. We measured the quality of surgery manuscripts selected by peer review and identified predictors of excellence. METHODS: One hundred twenty clinical surgery manuscripts were randomly selected from 1998 in 5 eminent peer reviewed surgery and medical journals. Manuscripts were blinded for author, institution, and journal of origin. Four surgeons and 4 methodologists evaluated the quality using novel instruments based on subject selection, study protocol, statistical analysis/inference, intervention description, outcome assessments, and results presentation. Predictors of quality and impact factor were identified using bivariate and multivariate regression. RESULTS: Oncology was the most common subject (26%), followed by general surgery/gastrointestinal (24%). The average number of study subjects was 417; the majority of manuscripts were American (53%), from a single institution (59%). Eighteen percent had a statistician author. Mean number of citations was 128. Surgery manuscripts from medical, compared with surgery journals, had better total quality scores (3.8 vs. 5.2, P < 0.001). They had more subjects and were more likely to have a statistician as coauthor (43% vs. 10%, P < 0.001), multi-institutional, international collaboration (30% vs. 8%, P < 0.001), and higher citation index (mean: 350 vs. 54, P < 0.001). They were more often foreign (70% vs. 40%, P < 0.001). Independent predictors of quality were having a statistician coauthor, study funding, European origin, and more study subjects. Quality assessment using our instruments predicted the number of citations after 10 years (P < 0.01), along with having a statistician coauthor, international multi-institutional collaboration, and more subjects. CONCLUSION: The quality of surgery manuscripts can be improved by including a statistician as coauthor, with efforts directed toward implementing multi-institutional/interdisciplinary trials. Peer-review across journals can be standardized through the use of instruments measuring methodologic and clinical quality. PMID- 19561472 TI - Hypothermia and surgery: immunologic mechanisms for current practice. AB - OBJECTIVE: To examine cellular and immunologic mechanisms by which intraoperative hypothermia affects surgical patients. SUMMARY BACKGROUND DATA: Avoidance of perioperative hypothermia has recently become a focus of attention as an important quality performance measure, aimed at optimizing the care of surgical patients. Anesthetized surgical patients are particularly at risk for hypothermia, which has been directly linked to the development of sequelae, such as coagulopathy, infection, morbid myocardial events, and death after surgery. However, many of the underlying immunologic mechanisms remain unclear. METHODS: Venous blood samples from healthy volunteers were exposed for up to 4 hours to various temperatures following the addition of a 1 ng/mL lipopolysaccharide challenge. Innate immune function, assessed by the ability of monocytes to present antigen and coordinate cytokine release, was determined by qualitative and quantitative measurements of HLA-DR surface expression 2 hours following incubation, and proinflammatory tumor necrosis factor-alpha (TNF-alpha) and anti inflammatory (IL-10) cytokine release in the first 4 hours. RESULTS: Monocyte incubation at hypothermic temperatures (34 degrees C) reduced HLA-DR surface expression, delayed TNF-alpha clearance, and increased IL-10 release. Conversely, hyperthermia (40 degrees C) increased monocyte antigen presentation and resulted in rapid decay of TNF-alpha. However, IL-10 release was also increased. Normothermia (37 degrees C) attenuated IL-10 release following the initial proinflammatory surge. CONCLUSION: Hypothermia exerts multiple effects at the cellular level, which impair innate immune function, and are associated with increased septic complications and mortality. These findings provide a physiological basis for perioperative temperature monitoring, which is a valid surgical performance measure that can be used to reduce surgical complications associated with avoidable hypothermia. PMID- 19561473 TI - Ghrelin hyporesponsiveness contributes to age-related hyperinflammation in septic shock. AB - OBJECTIVE: To test the hypothesis that hyporesponsiveness to ghrelin due to reduced growth hormone (GH) contributes to the aging-related hyperinflammatory state in sepsis. SUMMARY BACKGROUND DATA: Sepsis and septic shock are a serious problem, particularly in the geriatric population. Ghrelin is an endogenous ligand for the GH secretagogue receptor 1a (GHSR1a, ie, ghrelin receptor). The decline in GH with age is directly associated with many adverse changes that occur with aging. However, the role of GH, ghrelin, and GHSR1a in the age associated vulnerability to sepsis remains unknown. METHODS: Male Fischer 344 rats (young: 3 months; aged: 24 months) were used. Plasma GH levels, ghrelin receptor expression, and neuronal activity in the parasympathostimulatory nuclei of the brain stem in normal young and aged animals were measured. Endotoxemia was induced by intravenous injection of lipopolysaccharide (LPS, 15 mg/kg BW). RESULTS: While LPS-induced release of proinflammatory cytokines from macrophages isolated from aged rats decreased, LPS injection resulted in an in vivo hyperinflammatory state. GH levels were lower in aged rats, which was associated with lower expression of GHSR1a in the dorsal vagal complex and a decrease in parasympathostimulatory neuronal activity. GHSR1a antagonist elevated LPS-induced cytokine release in young rats. GH increased GHSR-1a expression in the dorsal vagal complex in aged rats. Coadministration of ghrelin and GH, but not ghrelin alone or GH alone, markedly reduced cytokine levels and organ injury after endotoxemia in aged rats, which was associated with significantly elevated parasympathostimulatory neuronal activity. CONCLUSIONS: These findings suggest that the reduced central (brain) responsiveness to ghrelin due to the decreased GH, plays a major role in producing the hyperinflammatory state, resulting in severe organ injuries and high mortality after endotoxemia in aged animals. Ghrelin and GH can be developed as a novel therapy for sepsis in the geriatric population. PMID- 19561474 TI - Prediction of postoperative outcome after hepatectomy with a new bedside test for maximal liver function capacity. AB - OBJECTIVE: To validate the LiMAx test, a new bedside test for the determination of maximal liver function capacity based on C-methacetin kinetics. To investigate the diagnostic performance of different liver function tests and scores including the LiMAx test for the prediction of postoperative outcome after hepatectomy. SUMMARY BACKGROUND DATA: Liver failure is a major cause of mortality after hepatectomy. Preoperative prediction of residual liver function has been limited so far. METHODS: Sixty-four patients undergoing hepatectomy were analyzed in a prospective observational study. Volumetric analysis of the liver was carried out using preoperative computed tomography and intraoperative measurements. Perioperative factors associated with morbidity and mortality were analyzed. Cutoff values of the LiMAx test were evaluated by receiver operating characteristic. RESULTS: Residual LiMAx demonstrated an excellent linear correlation with residual liver volume (r = 0.94, P < 0.001) after hepatectomy. The multivariate analysis revealed LiMAx on postoperative day 1 as the only predictor of liver failure (P = 0.003) and mortality (P = 0.004). AUROC for the prediction of liver failure and liver failure related death by the LiMAx test was both 0.99. Preoperative volume/function analysis combining CT volumetry and LiMAx allowed an accurate calculation of the remnant liver function capacity prior to surgery (r = 0.85, P < 0.001). CONCLUSIONS: Residual liver function is the major factor influencing the outcome of patients after hepatectomy and can be predicted preoperatively by a combination of LiMAx and CT volumetry. PMID- 19561475 TI - Polycystic liver disease: a critical appraisal of hepatic resection, cyst fenestration, and liver transplantation. AB - OBJECTIVE: To identify operative morbidity, mortality, and long-term outcome after operative treatment for symptomatic polycystic liver disease (PLD) and develop a treatment algorithm for patients with PLD. BACKGROUND: PLD represents a challenging clinical problem that can result in massive hepatomegaly and various complications, leading to significant decline in health status and quality of life. The optimal surgical treatment for this disease is still evolving. METHODS: All patients who underwent hepatic resection, cyst fenestration, or liver transplantation for PLD from 1985 to 2006 were identified retrospectively. Long term outcomes were evaluated by patient survey. Mean follow-up was 8 +/- 0.5 years. RESULTS: Of 141 patients (122 women; age: 51 +/- 1 years) with PLD, 117 had concomitant polycystic kidney disease. All patients suffered from symptomatic hepatomegaly with 85% being functionally impaired (Eastern Cooperative Oncology Group Performance Status: 1-3). Despite significant inferior vena cava or hepatic venous compression in 65%, hepatic function was commonly preserved. A total of 124 patients underwent partial hepatectomy with cyst fenestration, 10 underwent cyst fenestration alone, and 7 underwent liver transplantation for primary treatment of PLD. Overall operative morbidity and mortality was 58% and 4%, respectively, with major complications (Clavien grade: III-V) in 30%. Five- and 10-year survival was 90% and 78%, respectively. Eastern Cooperative Oncology Group Performance Status performance status normalized or improved in 75% of patients and 73% returned to work full-time. At follow-up, health survey scores were similar to the general population despite subsequent recurrence of symptoms in 73% of patients. CONCLUSION: Selective patients with massive hepatomegaly from PLD benefit from operative intervention. The type of operation performed is mainly dependent on the distribution of the cysts, coincident sectoral vascular patency and parenchymal preservation, and hepatic reserve. Hepatic resection can be performed with acceptable morbidity and mortality, prompt and durable relief of symptoms, and maintenance of liver function. Cyst fenestration and liver transplantation, though effective in selected patients, are less broadly applicable. PMID- 19561476 TI - Laparoscopic liver resection-understanding its role in current practice: the Henri Mondor Hospital experience. AB - OBJECTIVE: To report our complete experience with laparoscopic liver resection (LLR) to understand what role it may play in the broader context of liver surgery. BACKGROUND: The goal of LLR is to extend the benefits of the laparoscopic approach without compromising the fundamental principles of open liver surgery. LLR, however, presents unique technical challenges and its evaluation is made difficult by the restricted indications for this approach, the few centers world-wide experienced in the technique, and the heterogeneity of procedures and pathologies involved. METHODS: Retrospective analysis of a prospectively maintained database of liver resections from a unit with a comprehensive liver program, including resection and transplantation. RESULTS: There were 166 laparoscopic liver resections between May 23, 1996 and December 31, 2007, including 100 (60%) for malignant pathology (64 HCC, 3 cholangiocarcinoma, 33 hepatic metastases) and 66 for benign pathology (adenoma, 23; FNH, 19; cystic, 17; other, 7). Numbers of resections for benign indications remained stable over time whereas those for malignant indications increased. There were 31 major resections, 56 left lateral sectionectomies, 28 segmentectomies, and 51 tumorectomies. There was 0% mortality and 15.1% morbidity. Median blood loss was 200 mL, 9 patients (5.4%) required transfusion, and median operating time was 180 minutes. Left lateral sectionectomies demonstrated reduced bleeding (median, 175 vs. 300 mL, P = 0.0015) and faster operating time (median, 170 vs. 180 minutes, P = 0.0265). In the second half of the experience, there was reduced bleeding (median, 200 vs. 300 mL, P = 0.0022) and a lower conversion rate (2.4% vs. 16.9%, P = 0.0015). CONCLUSIONS: Good patient selection and refined surgical technique are the keys to successful LLR. The indications for resection of asymptomatic benign lesions should not be increased because the laparoscopic approach is available. Hepatocellular carcinomas (HCCs) are more likely to be suitable to a laparoscopic approach than colorectal liver metastases. Left lateral sectionectomy and limited resection of solitary peripheral lesions are particularly suitable while hemihepatectomies remain challenging procedures. LLR requires an ongoing robust audit to identify any emerging problems. PMID- 19561477 TI - Feasibility and efficacy of combination therapy with preoperative full-dose gemcitabine, concurrent three-dimensional conformal radiation, surgery, and postoperative liver perfusion chemotherapy for T3-pancreatic cancer. AB - OBJECTIVE: To evaluate both the feasibility and efficacy of our combined therapy, which consisted of preoperative chemoradiation, surgery, and postoperative liver perfusion chemotherapy (LPC) for patients with T3 (extended beyond the pancreatic confines) cancer of the pancreas. SUMMARY BACKGROUND DATA: Because of the high incidence of local recurrence and liver metastasis, long-term outcomes for patients after resection of T3-pancreatic cancer are extremely poor. METHODS: During the period from 2002 to 2007, 38 patients with T3-pancreatic cancers consented to receive a combination of preoperative chemoradiation, surgery, and postoperative LPC. With the aid of 3D radiation planning, irradiation fields were constructed that included both the primary pancreatic tumor and retropancreatic tissues while taking care to exclude any section of the gastrointestinal tract. The total dose of radiation was 50 Gy (2 Gy x 25 fractions/5 weeks) and was administered in combination with gemcitabine treatments (1000 mg/m/week x 9/3 months). Preoperative restaging via computerized tomography and intraoperative inspection were used to determine if pancreatectomy was indicated. For respected cases, one catheter was placed into the gastroduodenal artery and another one into the superior mesenteric vein. Postoperatively, 5-FU (125 mg/day x 28 days) was infused via each of these 2 routes. RESULTS: Preoperative chemoradiation was completed for all 38 patients, including 3 patients who required gemcitabine-dose reduction. Seven patients (18%) did not undergo surgical resection because either distant metastases or progressive local tumors had been detected after chemoradiation. The remaining 31 patients (82%) underwent pancreatectomy plus postoperative LPC, without postoperative or in-hospital mortality. The 5-year survival rate after pancreatectomy was 53%, with low incidences of both local recurrence (9%) and liver metastasis (7%). Postoperative histopathologic study revealed a marked degenerative change in cancer tissue, showing negative surgical margins (R0) for 30 patients (96%) and negative nodal involvement for 28 patients (90%). CONCLUSION: Results of this trial suggest that a combination of preoperative full-dose gemcitabine, concurrent 3D-conformal radiation, surgery, and postoperative LPC is feasible for the treatment of T3-pancreatic cancer. Using the method described in this article, we were able to effectively reduce the incidence of both local and liver recurrence. Therefore, this type of combination therapy seems promising for improving long-term outcomes for patients with T3-cancers of the pancreas. This study is registered with University hospital Medical information Network clinical trials Registry number, UMIN000001804. PMID- 19561478 TI - Multivisceral resection for pancreatic malignancies: risk-analysis and long-term outcome. AB - OBJECTIVE: To evaluate the safety and outcome of multivisceral pancreatic resections for primary pancreatic malignancies. BACKGROUND: Curative resection is the only potential cure for patients with pancreatic cancer, but some patients present with advanced tumors that are not resectable by a standard pancreatic resection. Data on risk and survival analysis of extended pancreatic resections is limited. METHODS: One hundred one patients who had a multivisceral pancreatic resection between 10/2001 and 12/2007 were identified from a prospective database, and perioperative and long-term results were compared with those of 202 matched patients with a standard pancreatic resection. Uni- and multivariate regression analysis were performed to identify parameters that are associated with perioperative morbidity. Long-term survival was evaluated. RESULTS: Colon, stomach, adrenal gland, liver, hepatic or celiac artery, kidney, or small intestine were resected in 37.6%, 33.7%, 27.7%, 18.8%, 16.8%, 11.9%, and 6.9% of the 101 patients with multivisceral resection, respectively. Additional portal vein resection was performed in 20.8% of patients. Overall and surgical morbidity but not mortality was significantly increased compared with standard pancreatic resections (55.5% vs. 42.8%, 37.6 vs. 25.3%, and 3.0% vs. 1.5%, respectively). Uni- and multivariate analysis identified a long operative time and the extended multivisceral resection of 2 or more additional organs as independent risk factors for intraabdominal complications or need for relaparotomy. Median survival was comparable to that of standard pancreatic resections. CONCLUSIONS: Multivisceral resections can be performed with increased morbidity but comparable mortality and long-term prognosis as compared with standard pancreatic resections at high volume centers. Increased morbidity is related to extended multivisceral resections with a long operative time. PMID- 19561479 TI - Survival after pancreaticoduodenectomy is not improved by extending resections to achieve negative margins. AB - OBJECTIVE: This study was undertaken to determine the survival benefit of extending resections to obtain microscopically negative margins after positive intraoperative frozen sections. SUMMARY BACKGROUND DATA: The impact of residual microscopic disease after pancreaticoduodenectomy is currently a point of controversy. It is, however, generally believed that microscopically positive margins negatively impact survival and this may be improved by ultimately achieving negative margins. METHODS: Since 1995, patients undergoing pancreaticoduodenectomy for pancreatic adenocarcinoma have been prospectively followed. Margin status has been codified as macro/microscopically negative (R0) or macroscopically negative/microscopically positive (R1). The impact of margin status on survival was evaluated utilizing survival curve analysis. Data are presented as median, mean +/- SD where appropriate. RESULTS: For pancreatic adenocarcinoma, 202 patients underwent pancreaticoduodenectomy. R0 resections were achieved in 158 patients, 17 of whom required extended resections to achieve complete tumor extirpation after an initially positive intraoperative frozen section (R1 --> R0). R1 resections were undertaken in 44 patients. Median survival for patients undergoing R0 resections was 21 months, 26 +/- 23.4 months versus 13 months, 17 +/- 21.0 months for patients undergoing R1 resections (P = 0.02). Median survival for patients undergoing R1 --> R0 resections was 11 months, 16 +/- 17.3, (P = 0.001). Margin status had a significant correlation with "N" stage and AJCC stage but not "T" stage. CONCLUSION: Survival after pancreaticoduodenectomy is not improved by extending pancreatic resections to achieve negative margins after initially positive intraoperative frozen sections. Tumor-specific factors beyond the presence of disease at a surgical margin are responsible for the abbreviated survival seen in patients undergoing R1 resections. PMID- 19561480 TI - Career decisions and the structure of training: an American Board Of Colon and Rectal Surgery survey of colorectal residents. AB - OBJECTIVE: To investigate potential impacts of restructuring general surgery training on colorectal (CR) surgery recruitment and expertise. SUMMARY BACKGROUND DATA: In response to the American Surgical Association Blue Ribbon Committee report on surgical education (2004), the American Board of Colon and Rectal Surgery, working with the Accreditation Council for Graduate Medical Education and American Board of Surgery, established a committee (2006) to review residency training curricula and study new pathways to certification as a CR surgeon. To address concerns related to shortened general surgery residency, the American Board of Colon and Rectal Surgery committee surveyed recent, current, and entering CR residents on the timing and factors associated with their career choice and opinions regarding restructuring. METHODS: A 10-item, online survey of 189 CR surgeons enrolled in the class years of 2005, 2006, and 2007 was administered and analyzed May to July 2007. RESULTS: One hundred forty-five CR residents responded (77%); results were consistent across class years and types of general surgery training program. Seventy percent of respondents had rotated onto a CR service by the end of their PGY-2 year. Most identified CR as a career interest in their PGY-3 or PGY-4 year. Overall interest in CR surgery, the influence of CR mentors and teachers, and positive exposure to CR as PGY-3, PGY 4, or PGY-5 residents were the top cited factors influencing choice decisions. Respondents were opposed to restructuring by a 2:1 ratio, primarily because of concerns about inadequate training and lack of time to develop technical expertise. CONCLUSIONS: Shortening general surgery residency would not necessarily limit exposure to CR rotations and mentors unless such rotations are cut. The details of proposed restructuring are critical. PMID- 19561481 TI - Laparoscopic versus open surgery for rectal cancer: long-term oncologic results. AB - OBJECTIVE: The goal was to assess long-term oncologic outcome after laparoscopic versus open surgery for rectal cancer and to evaluate the impact of conversion. SUMMARY BACKGROUND DATA: Laparoscopic resection of rectal cancer is technically feasible, but there are no data to evaluate the long-term outcome between laparoscopic and open approach. Moreover, the long-term impact of conversion is not known. METHODS: Between 1994 and 2006, patients treated by open (1994-1999) and laparoscopic (2000-2006) curative resection for rectal cancer were included in a retrospective comparative study. Patients with fixed tumors or metastatic disease were excluded. Those with T3-T4 or N+ disease received long course preoperative radiotherapy. Surgical technique and follow-up were standardized. Survival were analyzed by Kaplan Meier method and compared with the Log Rank test. RESULTS: Some 471 patients had rectal excision for invasive rectal carcinoma: 238 were treated by laparoscopy and 233 by open procedure. Postoperative mortality (0.8% vs. 2.6%; P = 0.17), morbidity (22.7% vs. 20.2%; P = 0.51), and quality of surgery (92.0% vs. 94.8% R0 resection; P = 0.22) were similar in the 2 groups. At 5 years, there was no difference of local recurrence (3.9% vs. 5.5%; P = 0.371) and cancer-free survival (82% vs. 79%; P = 0.52) between laparoscopic and open surgery. Multivariate analysis confirmed that type of surgery did not influence cancer outcome. Conversion (36/238, 15%) had no negative impact on postoperative mortality, local recurrence, and survival. CONCLUSIONS: The efficacy of laparoscopic surgery in a team specialized in rectal excision for cancer (open and laparoscopic surgery) is suggested with similar long-term local control and cancer-free survival than open surgery. Moreover, conversion had no negative impact on survival. PMID- 19561482 TI - Bowel obstruction following appendectomy: what is the true incidence? AB - INTRODUCTION: Appendicitis is a common problem that is typically treated with an appendectomy. Following abdominal surgery, adhesions may form and may cause a subsequent small bowel obstruction (SBO). The purpose of our study was to determine the rate of post-appendectomy SBO in an adult population, and to observe any difference in SBO rates between open versus laparoscopic appendectomies. METHODS: All patients who underwent an appendectomy at an adult hospital in the Calgary Health Region between 1999 and 2002 were identified by using the administrative discharge database. Pathology and operative technique (laparoscopic, McBurney incision, midline laparotomy) were reviewed. Using those regional health numbers, any further admissions with a diagnostic code for bowel obstruction were identified. Medical charts (n = 1777) were reviewed to confirm the rate of post-appendectomy SBO. A logistic regression was performed to identify risk factors of post-appendectomy SBO and expressed as odds ratios (95% confidence interval). RESULTS: The overall SBO rate was 2.8% over an average 4.1 year follow-up period. The risk factors for developing SBO following appendectomy for appendicitis included, perforated appendicitis (odds ratio [OR] = 3.1, 95% confidence interval [CI]: 1.5-6.6), and midline incisions (OR = 5.4, 95% CI: 2.8 10.4). Those with pathology of cancer or chronic appendicitis conferred the greatest overall risk of SBO (OR = 7.4, 95% CI: 2.7-20.3). CONCLUSIONS: The rate of SBO following appendectomy in adults was 2.8%, or 0.0069 cases per person year. The greatest risk factors for developing SBO were midline incision and nonappendicitis pathology. There is no statistically significant difference in SBO rates following laparoscopic appendectomy compared with open approaches. PMID- 19561483 TI - Different pathological features and prognosis in gastric cancer patients coming from high-risk and low-risk areas of Italy. AB - OBJECTIVE: To compare clinicopathological features and long-term outcome in gastric cancer patients coming from high-risk and low-risk areas of Italy. SUMMARY BACKGROUND DATA: Better survival rates have been reported from countries with higher incidence of gastric cancer. METHODS: Data regarding 829 patients coming from Tuscany (group A) and 143 patients coming from Southern Italy (group B) were analyzed. Mean follow-up time was 56 +/- 57 months; it was 85 +/- 63 months in surviving patients or not tumor-related deaths. Prognostic factors were investigated by multivariate analysis with Cox proportional hazard model after verifying the assumption of proportionality of the risk associated with covariates. RESULTS: Lauren diffuse-mixed histotype, younger age, extended lymphadenectomy, and advanced stages were more common in group B. Gastric cancer related 10-year survival probability was 48% in group A versus 29% in group B (log-rank test: P < 0.001). By multivariate analysis, geographic area was confirmed as a significant prognostic factor (hazard ratio for group B vs. group A: 1.52, 95% confidence interval: 1.12-2.06, P = 0.006). The influence of this factor on long-term survival was independent from other clinical, surgical, and pathologic factors, and was notable in neoplasms involving the serosa (10-year survival probability: 15% in group A vs. 3% in group B, log-rank test: P = 0.005). CONCLUSIONS: Patients coming from low-risk area of Italy showed distinct pathologic features, more advanced stage, and worse prognosis when compared with patients coming from high-risk area. These findings may be indicative of different tumor biology, and may contribute to partly explain worldwide geographic variability in prognosis reported in different series. PMID- 19561484 TI - Open mesh techniques for inguinal hernia repair: a meta-analysis of randomized controlled trials. AB - OBJECTIVE: The aim of this report is to evaluate the efficacy of the 3 most common open mesh techniques used in practice. BACKGROUND: The open mesh technique of choice, inguinal hernia repair, remains controversial. Some randomized controlled trials (RCTs) have been published assessing its effectiveness; however, available data regarding the outcomes were noncoincident. Therefore, it is necessary to carry out a meta-analysis of the evidence on outcomes for the open mesh technique. METHODS: : A fully recursive literature search was conducted in the Cochrane Controlled Trials Register Databases, Medline, Embase, ISI databases, and Chinese Biomedical Literature Database in any language. RCTs between 1989 and 2008 of open mesh techniques were considered for inclusion. Statistical analyses were carried out using RevMan software. RESULTS: Ten RCTs with 2708 patients were included. There was no significant difference in recurrence when comparing Lichtenstein's operation to mesh plug repair or prolene hernia system (PHS) repair (risk ratio: 0.71; 95% confidence intervals: 0.32 1.56), (risk ratio: 2.19; 95% confidence intervals: 0.63-7.62), respectively. There was insufficient evidence to determine the greater effectiveness between mesh plug repair and PHS repair. CONCLUSIONS: Lichtenstein's operation was similar to mesh plug or PHS repair in terms of time to return to work, complications, chronic pain, and hernia recurrence in the short- to mid-term, although mesh plug and PHS repair was associated with shorter surgical time. PMID- 19561485 TI - Preliminary results of a prospective randomized trial of restrictive versus standard fluid regime in elective open abdominal aortic aneurysm repair. AB - BACKGROUND: Open abdominal aortic aneurysm (AAA) repair is associated with a significant morbidity (primarily respiratory and cardiac complications) and an overall mortality rate of 4% to 10%. We tested the hypothesis that perioperative fluid restriction would reduce complications and improve outcome after elective open AAA repair. METHODS: In a prospective randomized control trial, patients undergoing elective open infra-renal AAA repair were randomized to a "standard" or "restricted" perioperative fluid administration group. Primary outcome measure was rate of major complications (MC) after AAA repair and secondary outcome measures included: Sequential Organ Failure Assessment Score; FiO2/PO2 ratio; Urinary Albumin/Creatinine Ratio; Length-of-stay in, intensive care unit, high dependency unit, in-hospital. This prospective Randomized Controlled Trial was registered in a publicly accessible database and has the following ID number ISRCTN27753612. RESULTS: Overall 22 patients were randomized, 1 was excluded on a priori criteria, leaving standard group (11) and restricted group (10) for analysis. No significant difference was noted between groups in respect to age, gender, American Society Anesthesiology class, Physiological and Operative Severity Score for the Enumeration of Mortality and Morbidity scores, operation time, and operation blood loss. There were no in-hospital deaths and no 30-day mortality. The cumulative fluid balance on day 5 postoperative was for standard group, 8242 +/- 714 mL, compared with restricted group, 2570 +/- 977 mL, P < 0.01. MC were significantly reduced in the restricted group (n = 10), 1 MC, compared with standard group (n = 11), 14 MC, P < 0.024. Total and postoperative length-of-stay in-hospital was significantly reduced in the restricted group, 9 +/- 1 and 8 +/- 1 days, compared with standard group, 18 +/- 5 and 16 +/- 5 days, P < 0.01 and P < 0.025, respectively. CONCLUSIONS: Serious complications are common after elective open AAA repair, and we have shown for the first time that a restricted perioperative fluid regimen can prevent MC and significantly reduce overall hospital stay. PMID- 19561487 TI - Timing is everything. PMID- 19561489 TI - Relative contributions of collagen and tissue factor to thrombus formation on damaged vascular vessels: in-vitro studies with circulating blood. AB - BACKGROUND AND OBJECTIVE: Thrombogenicity of ruptured atherosclerotic plaques is mainly attributed to the exposure of collagen (Col) and tissue factor (TF). Using in-vitro approaches, we explored the relative contribution of Col and TF to local thrombogenesis. METHODS: Surfaces coated with Col and human TF, alone or in combinations (diluted Col on TF, and diluted TF on Col), were exposed to flowing blood at a shear rate of 600/s. Platelet and fibrin deposition were analyzed morphometrically. Generation of prothrombin fragments (F1+2) was determined as a measure of global activation of coagulation. RESULTS: Col and TF alone behaved as adhesive substrata to platelets supporting similar platelet coverage around 22%. Col induced tight aggregates, whereas TF promoted adhesive events. In studies with combinations of Col and TF, platelet aggregation was always enhanced with statistical elevations of the thrombus area (P<0.05). Generation of F1+2 was surface-dependent and was at its highest levels when both proteins were combined (P<0.05). However, local fibrin formation was statistically increased in surfaces containing lower concentrations of TF on Col (P<0.01 vs. overall surfaces assessed). CONCLUSION: Our studies show that combinations of Col and TF always enhance thrombogenesis, but small amounts of TF on Col surfaces resulted in the most procoagulant combination. The present results suggest that plaques exposing high contents of Col, with small amounts of TF, would provide the most occlusive combination. PMID- 19561488 TI - Involvement of N-acetyltransferase human in the cytotoxic activity of 5 fluorouracil. AB - N-acetyltransferase human (NATH) participates in a posttranslational modification of the proteins and has been reported to play a role in apoptosis. In this study, the involvement of NATH in the cytotoxic action of 5-fluorouracil (5-FU) in human squamous cell carcinoma HEp-2 cells was examined. We found that 5-FU decreased NATH expression in a dose-dependent and time-dependent manner. No change was observed after treatment with bleomycin, nedaplatin, mitomycin C, or methotrexate. Interestingly, knockdown of NATH by small interfering RNA resulted in the downregulation of thymidylate synthase mRNA expression and induced apoptosis. Conversely, NATH overexpression facilitated cell proliferation independent of the presence of 5-FU. The effect of NATH knockdown on the expression of proteins in HEp-2 cells was examined using two-dimensional gel electrophoresis and mass spectrometry. Profilin 1, CutA, ras-related nuclear protein, annexin A5, enolase 1, and elongation factor 1 alpha 1 were found to be upregulated and 14-3-3eta, tublin, nuclear auto antigenic sperm protein, heat shock protein 70, and heat shock protein 90 were downregulated by knockdown of NATH. The results of this study suggest that NATH plays an important role in the cytotoxic activity of 5-FU. PMID- 19561486 TI - Timing of antimicrobial prophylaxis and the risk of surgical site infections: results from the Trial to Reduce Antimicrobial Prophylaxis Errors. AB - OBJECTIVE: The objective of this study is to determine the optimal timing for surgical antimicrobial prophylaxis (AMP). SUMMARY BACKGROUND DATA: National AMP guidelines should be supported by evidence from large contemporary data sets. METHODS: Twenty-nine hospitals prospectively obtained information on AMP from 4472 randomly selected cardiac, hip/knee arthroplasty, and hysterectomy cases. Surgical site infections (SSIs) were ascertained through routine surveillance, using National Nosocomial Infections Surveillance system methodology. The association between the prophylaxis timing and the occurrence of SSI was assessed using conditional logistic regression (conditioning on hospital). RESULTS: One hundred thirteen SSI were detected in 109 patients. SSI risk increased incrementally as the interval of time between antibiotic infusion and the incision increased (overall association between timing and infection risk P = 0.04). When antibiotics requiring long infusion times (vancomycin and fluoroquinolones) were excluded, the infection risk following administration of antibiotic within 30 minutes prior to incision was 1.6% compared with 2.4% associated with administration of antibiotic between 31 to 60 minutes prior to surgery (OR: 1.74; 95% confidence interval, 0.98-3.04). The infection risk increased as the time interval between preoperative antibiotic and incision increased or if the antibiotic was first infused after incision. Intraoperative redosing (performed in only 21% of long operations) appeared to reduce SSI risk in operations lasting more than 4 hours (OR of 3.08 with no redosing; 95% confidence interval 0.74-12.90), but only when the preoperative dose was given correctly. CONCLUSIONS: These data from a large multicenter collaborative study confirm and extend previous observations and show a consistent relationship between the timing of AMP and SSI risk with a trend toward lower risk occurring when AMP with cephalosporins and other antibiotics with short infusion times were given within 30 minutes prior to incision. PMID- 19561490 TI - Selective immediate hypersensitivity reactions to NSAIDs. AB - PURPOSE OF REVIEW: Selective immediate reactions to NSAIDs imply that patients develop a urticarial/anaphylactic response to a single drug with good tolerance to other compounds. No systematic review of these reactions has yet been made. RECENT FINDINGS: With the increase in consumption of NSAIDs, these have become one of the most common drugs inducing hypersensitivity reactions. Although cross intolerance reactions are the most common, a significant proportion is selective responses. As specific IgE antibodies are not always found, there is only indirect evidence supporting an IgE-mediated mechanism in selective NSAID reactors. SUMMARY: Selective immediate reactions to NSAIDs must be considered when a patient develops urticaria or anaphylaxis after intake of one drug with good tolerance to drugs from other groups or even a drug from the same group with a slightly different chemical structure. Further research is required to identify the antigenic determinant structures recognized. PMID- 19561492 TI - New treatments for methicillin-resistant Staphylococcus aureus. AB - PURPOSE OF REVIEW: Methicillin-resistant Staphylococcus aureus (MRSA) is a dynamic pathogen. Rates of MRSA are increasing worldwide. In some centers, MRSA is becoming less susceptible to vancomycin, and these strains have been associated with worse clinical outcomes. Intermediate or fully resistant vancomycin strains of MRSA have emerged clinically, whereas MRSA acquired in the community has become epidemic. The purpose of this manuscript is to provide clinicians with an evidence-based review on new treatments for MRSA. RECENT FINDINGS: Linezolid, daptomycin and tigecycline have been approved during the last decade to treat infections due to MRSA. Although these agents are extremely valuable in the fight against MRSA, each one has limitations. New lypoglycopeptides (telavancin, dalbavancin and oritavancin) are in advanced phase of clinical development. Similarly, new broad-spectrum cephalosporins active against MRSA (e.g. ceftobiprole and ceftaroline) and a new dihydrofolate reductase inhibitor (iclaprim) are in or have completed phase 3 studies. SUMMARY: Here, we review the most relevant information on new drugs to treat MRSA. New studies with available agents and upcoming studies with investigational drugs will help to better understand the role of each compound in the treatment of patients infected with MRSA and assist the clinician in keeping pace with this challenging pathogen. PMID- 19561493 TI - A systems approach to the early recognition and rapid administration of best practice therapy in sepsis and septic shock. AB - PURPOSE OF REVIEW: The early recognition and treatment of sepsis is paramount to reducing the mortality of this disease. However, unlike trauma, stroke or acute myocardial infarction, the initial signs of sepsis are subtle and easily missed by clinicians. Thus, hospital-based systems are needed to identify and triage patients who might be septic. This review focuses on the early diagnosis of sepsis and the implementation of a systems-based approach to help coordinate the identification and treatment of patients with this disease. RECENT FINDINGS: Alterations in traditional hemodynamic parameters, such as blood pressure and heart rate, are poor predictors of the presence of septic shock. Other more subtle findings (such as the 10 signs of vitality) are stronger determinants of poor tissue perfusion in a patient who may be septic. Early detection of a patient who is 'in trouble' on the ward by bedside nurses or physicians and activation of a medical emergency team has been shown to improve outcome. By coupling the medical emergency team with early goal-directed therapy, patients with sepsis can be discovered earlier and have therapy instituted within the so called 'golden hour', first appreciated with trauma care. SUMMARY: The institution of a rapid response system for the detection and treatment of septic shock requires a multidisciplinary approach. The infrastructure to create such a system must be facilitated by administrators and implemented by front-line healthcare providers. Continuous assessment of the outcome benefit of such a system by a quality assurance team is the final part of a truly integrated approach to sepsis treatment. PMID- 19561494 TI - Should proteinuria reduction be the criterion for antihypertensive drug selection for patients with kidney disease? AB - PURPOSE OF REVIEW: Proteinuria, that is, more than 200 mg/day of urinary albumin, is associated with the presence of kidney disease. Its increase over time is strongly correlated with progression of nephropathy. Retrospective analyses of nephropathy outcome trials show that proteinuria reduction of 30% or more after initiation of blood pressure (BP)-lowering therapy is associated with slower nephropathy progression than lowering BP without its reduction. RECENT FINDINGS: Retrospective analyses of five large nephropathy outcome trials demonstrate that nephropathy progression slowed by an additional 28-39% over the control or placebo group when proteinuria was reduced in concert with BP. Two separate trials demonstrate that nephropathy progression was slowed to a lesser degree when BP was reduced to a similar degree, but proteinuria reduced less than 30%. These associations do not hold for those with microalbuminuria, in which BP reduction is the key element to slowing nephropathy progression. Recent cardiovascular outcome trials fail to show a relationship between reductions in proteinuria and nephropathy outcomes. This large cardiovascular endpoint trial, however, was not only powered for nephropathy outcomes but also failed to show a benefit between proteinuria reduction and cardiovascular events, a previously established observation. SUMMARY: All patients with a history of hypertension and either kidney disease or diabetes should have an annual check for albuminuria. If albumin is present in amounts of more than 200 mg/day, strategies for BP-lowering therapy should also focus on a reduction of more than 30% of urinary protein. PMID- 19561495 TI - Should diuretics always be included as initial antihypertensive management in early-stage CKD? AB - PURPOSE OF REVIEW: This review focuses on the need for combined antihypertensive therapy drugs in patients with chronic kidney disease and the relevance of diuretics. RECENT FINDINGS: Chronic kidney disease is a high-risk situation characterized by the presence of volume overload-related hypertension, micro/macroalbuminuria and other traditional and nontraditional risk factors. To achieve the blood pressure goal in these patients, combined antihypertensive therapy (including diuretics) is usually required. SUMMARY: Extracellular volume expansion is an important, if not the most important, contributing factor to hypertension seen in chronic kidney disease. Protection against progression of renal dysfunction has two main requirements: strict blood pressure control and lowering proteinuria to values as near to normal as possible. Diuretics have been a useful tool to manage volume overload and to achieve strict blood pressure control in patients with chronic kidney disease. Albeit other blood pressure lowering agents offer additional favorable effects independently of blood pressure changes, diuretics will continue to be used in these patients. PMID- 19561496 TI - Cellular physiology of the renal H+ATPase. AB - PURPOSE OF REVIEW: Vacuolar-type H+ATPases are multisubunit macromolecules that play an essential role in renal acid-base homeostasis. Other cellular processes also rely on the proton pumping ability of H+ATPases to acidify organellar or lumenal spaces. Several diseases, including distal renal tubular acidosis, osteoporosis and wrinkly skin syndrome, are due to mutations in genes encoding alternate subunits that make up the H+ATPase. This review highlights recent key articles in this research area. RECENT FINDINGS: Further insights into the structure, expression and regulation of H+ATPases have been elucidated, within the kidney and elsewhere. This knowledge may enhance the potential for future drug targeting. SUMMARY: Novel findings concerning tissue-specific subunits of the H+ATPase that are important in the kidney and more general lessons of H+ATPase function and regulation are slowly emerging, though the paucity of cellular tools available has to date limited progress. PMID- 19561497 TI - A paradigm shift: volume augmentation or 'inflation' to obtain optimal cosmetic results. AB - PURPOSE OF REVIEW: In the last several years, surgeons have increasingly realized the importance of volume loss as a central mechanism of facial aging. New, innovative techniques incorporate volume augmentation, or inflation, to improve cosmetic outcomes. RECENT FINDINGS: Previous rejuvenative facial surgical techniques focused on the removal of excess, lax tissues. In addition to tissue laxity and descent, volume loss significantly contributes to the appearance of facial aging in most patients. Volume loss, or deflation, affects all facial layers, including the skin, deep soft tissue, and bone. A variety of new, innovative surgical and nonsurgical techniques specifically address volume deficiencies in the brow, periorbita, and midface. Volume augmentation directly counteracts volume loss, a key feature of facial aging, and offers surgeons a powerful tool to improve cosmetic results. SUMMARY: In addition to the excision or repositioning of lax or prolapsed tissues, surgeons must assess and treat facial volume loss to optimize cosmetic outcomes. PMID- 19561498 TI - Surgical management of acquired socket contracture. AB - PURPOSE OF REVIEW: In a cosmetically oriented world, the inability to retain an ocular prosthesis remains a psychologically devastating problem for patients. This study reviews the recent advances in the management of acquired socket contracture. RECENT FINDINGS: For mild contracture, several recent studies describe the beneficial use of amniotic membrane as a substrate graft to allow the conjunctival epithelium to migrate and multiply. To correct moderately severe socket contracture, quilting sutures are typically used to ensure survival of mucous membrane grafts, which are required to provide adequate lining to the socket. In cases of severe socket contraction, irradiated sockets, and recurrent socket contraction, radial-free forearm flaps or a variety of other flaps in association with bone expansion osteotomies are on the rise. SUMMARY: Techniques to treat contracted sockets continue to evolve; however, large multicenter trials as well as an updated classification scheme are needed to devise standardized treatment protocols. PMID- 19561499 TI - The use of osseointegration and rare earth magnetic coupling for oculofacial prosthesis retention in the exenterated orbit. AB - PURPOSE OF REVIEW: The goal of the present review is to update ophthalmologists and oculoplastic surgeons with the role of osseointegration for oculofacial prosthesis retention in patients requiring orbital exenteration surgery. RECENT FINDINGS: The biophysiology of osseointegration has been well established by our dentistry colleagues and is reviewed here. Using this technology, orbital surgeons can offer their patients an adhesive-free method of prosthesis retention. Recent literature discusses the difficulties of implant success in the irradiated orbit. SUMMARY: Osseointegration with rare earth magnetic coupling provides patients with proper oculofacial prosthesis alignment and retention assisting in the rehabilitation of the exenterated orbit. PMID- 19561500 TI - Facial nerve schwannoma. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the current literature on facial nerve schwannoma and make practical recommendations based on best practices for the management of this difficult but benign neoplasm. RECENT FINDINGS: Facial nerve schwannoma can be asymptomatic or can present with progressive or acute facial nerve palsy. Associated otological symptoms such as conductive and/or sensorineural hearing loss can occur. The tumor is usually slow growing and can involve multiple segments of the nerve. Radiographic imaging and facial nerve electrical testing can be helpful in treatment planning. Options for management can include observation, decompression, stripping, resection with grafting, and possibly radiotherapy. Future adjunctive therapies to improve facial nerve function may include electrical stimulation, steroid hormones, and possibly stem cell therapy. SUMMARY: Treatment of facial schwannoma is individualized based on patient symptoms, history, and clinicoradiographic evaluation. Not all patients require surgery. As the tumor can involve multiple segments of the nerve, the surgeon attempting removal should be familiar with modern neurotological surgical techniques. Ongoing translational research will hopefully allow us to decrease facial nerve morbidity in these patients. PMID- 19561501 TI - Managing the psychological aspects of plastic surgery patients. AB - PURPOSE OF REVIEW: The success of any physician-patient interaction is based on good communication. This requires the physician to be a careful listener with clinical acumen that extends beyond the typical borders of medical illness. For the plastic surgeon, the skill set must include an ability to understand the psychological motivations of the patient for the given treatment and must allow the surgeon to put himself into the position of the patient for this acceptance. Additionally, the surgeon must decide whether the patient is a good physiological candidate for surgery. RECENT FINDINGS: There are many personality types that suggest problems postoperatively. These include patients who are overly demanding, highly critical, have poor self-image, and those with body dysmorphisms. SUMMARY: In order to have a successful result in plastic surgery, the surgeon must select his patients well, perform meticulous operative procedures, and give empathetic postoperative care. It is this combination of care that extends beyond aesthetic or functional disorders that will allow success in plastic surgery. PMID- 19561502 TI - Perspectives of intellectual disability in Latin American countries: epidemiology, policy, and services for children and adults. AB - PURPOSE OF REVIEW: The prevalence of intellectual disability is an estimated 1-4% worldwide. Etiological factors such as malnutrition, lack of perinatal care, and exposure to toxic and infectious agents, which are more common in low-income and middle-income (LAMI) countries, may contribute to a higher prevalence of intellectual disability in Latin America. This review summarizes the data on intellectual disability coming from Latin America, which is published in scientific journals and is available from official websites and discusses potential health policy and services implications of these studies. RECENT FINDINGS: Methodologically rigorous studies on intellectual disability in Latin America are lacking. This paucity of basic epidemiological information is a barrier to policy and services development and evaluation around intellectual disability. Only two studies, one from Chile and another from Jamaica, allow for adequate population estimates of intellectual disability. Interestingly, the countries with the highest scientific production in Latin America, Brazil and Mexico, did not produce the most informative research in epidemiology, policy or services related to intellectual disability. SUMMARY: The main conclusion of this review is that a lack of scientific evidence makes it difficult to properly characterize the context of intellectual disability in Latin America. Insufficient data is also a barrier to policy and services development for governments in Latin America. Although recently there have been efforts to develop government programs to meet the needs of the intellectual disability population in Latin America, the effectiveness of these programs is questionable without proper evaluation. There is a need for studies that characterize the needs of people with intellectual disability specifically in Latin America, and future research in this area should emphasize how it can inform current and future policies and services for people with intellectual disability. PMID- 19561503 TI - Special education for intellectual disability: current trends and perspectives. AB - PURPOSE OF REVIEW: To inform readers of current issues in special education for individuals with intellectual disabilities and summarize recent research and opinion. RECENT FINDINGS: Two issues dominate special education for students with intellectual disabilities in the early 21st century. First, what should be taught to such students and who should teach them? Second, where should such students be taught - in 'inclusive' settings alongside normal peers or in special settings dedicated to their special needs? Research on teaching reading, arithmetic, and functional daily living skills to students with disabilities suggests the superiority of direct, systematic instruction. Universal design is often seen as supportive of inclusion. Inclusion has been seen as the central issue in special education but is gradually giving way to concern for what students learn. SUMMARY: Direct, systematic instruction in reading, arithmetic, and daily living skills is the most effective approach to teaching students with intellectual disabilities. Basic concepts and logic suggest that special and general education cannot be equivalent. We conclude that what students are taught should be put ahead of where they are taught. Our fundamental concern is that students with intellectual disabilities be respected and be taught all they can learn. PMID- 19561504 TI - Perspectives of intellectual disability in Africa: epidemiology and policy services for children and adults. AB - PURPOSE OF REVIEW: To establish the nature of publications emanating from or relating to intellectual disability in Africa in the previous 12-18 months, and to critically analyse the nature and content of any new knowledge arising from them. RECENT FINDINGS: Three papers satisfied the criteria set, one from Nigeria on epilepsy and intellectual disability, the other two were from South Africa. Kromberg's paper puts intellectual disability in context for South Africa, emphasizing the role potentially played by traditional healers. Pillay's paper tackles emerging issues in the psycholegal arena in South Africa, many of which are relevant to the rest of the continent. SUMMARY: Of the many challenges facing persons with intellectual disability in Africa, high prevalence, discrimination, and access to justice and education are key considerations, in all countries, and without exception are coupled with poor resource allocation. The challenges posed by HIV, wars and internal displacement are significant and demand examination by researchers. Opportunities for beneficial research exist between Africa and the centres of academic excellence around the rest of the world. Those centres with the skill and will to collaborate with Africa will be handsomely rewarded by the wide scope of challenges in intellectual disability that are available for scientific enquiry. Opportunities for skills transfer are many and available. PMID- 19561506 TI - Alveolar proteinosis syndrome: pathogenesis, diagnosis, and management. AB - PURPOSE OF REVIEW: This review discusses the most recent clinical and basic research literature on pulmonary alveolar proteinosis (PAP) as it relates to pathogenesis, diagnosis, and management. RECENT FINDINGS: The discovery of Granulocyte macrophage-colony stimulating factor (GM-CSF) and the alveolar macrophage as critical regulators of surfactant protein and lipid homeostasis has led to significant advances in PAP. Adults affected by PAP have circulating neutralizing anti-GM-CSF antibodies. Reduced localized GM-CSF activity in the lung (from neutralizing anti-GM-CSF antibodies), decreases alveolar macrophage surfactant degradation with surfactant excess and accumulation. Cause, source of antibodies or downstream effects of GM-CSF deficiency is speculative. GM-CSF antibodies above a threshold level have proved to be a useful diagnostic test. Research towards therapy has focused on improving the technique for therapeutic whole lung lavage as well as overcoming effects of neutralizing anti-GM-CSF, which include GM-CSF therapy (systemic and inhaled) and anecdotal reports of anti B cell therapy. Whereas this approach has been somewhat successful for primary PAP, other causes of PAP (i.e. alveolar macrophage dysfunction, surfactant protein alterations) are still without therapy. SUMMARY: Understanding of the pathogenesis of PAP has greatly increased in the last decade; study has brought better comprehension of lung biology and recognition of the critical role for GM CSF and alveolar macrophage in surfactant clearance. Balance between resident immune cell population and normal lung function still needs further study. Resident alveolar macrophages have an essential role in surfactant homeostasis. With this knowledge more effective diagnostic tests (e.g. anti-GM-CSF antibody) and therapies for PAP are under investigation. PMID- 19561507 TI - New results from the use of bisphosphonates in cancer patients. AB - PURPOSE OF REVIEW: Metastatic bone disease results from cancer-bone cell interactions in the bone marrow microenvironment rather than direct destruction by cancer cells. Additionally, cancer treatments may have adverse effects on bone health. Therefore, a clear rationale exists for bone-targeted therapies to prevent skeletal complications and potentially modify the underlying disease. RECENT FINDINGS: Studies in metastatic disease have focused on refining treatment, largely through the use of bone metabolism markers. Use of bisphosphonates in early cancer has become increasingly important to prevent bone loss. Recent results, notably with zoledronic acid, indicate that bone mineral density can be maintained and increased bone turnover normalized. Bisphosphonates are potent inhibitors of bone osteolysis and may impede both the development of bone metastases and survival of dormant cells in the marrow microenvironment. Additionally, bisphosphonates may have direct effects on tumour cells, especially in combination with chemotherapy. Clinical trial results in breast cancer with clodronate were inconclusive, but recent data with zoledronic acid suggest that bisphosphonates may indeed modify the course of the disease. SUMMARY: The roles of bisphosphonates in metastatic bone disease and the prevention of cancer treatment induced bone loss have been defined. However, results from ongoing metastasis prevention trials are required before routine adjuvant bisphosphonates can be recommended. PMID- 19561505 TI - Chronic kidney disease and venous thromboembolism: epidemiology and mechanisms. AB - PURPOSE OF REVIEW: An estimated 13% of Americans have kidney disease. We sought to describe the association of kidney disease with risk of venous thromboembolism and discuss possible mechanisms explaining this association. RECENT FINDINGS: All severities of kidney disease appear to increase the risk of venous thromboembolism. In the general population the risk associated with mild to moderate kidney disease is 1.3-2-fold increased, and present even for microalbuminuria, although stage 1 chronic kidney disease itself has not been studied. End-stage renal disease is also associated with a 2.3-fold increased risk, compared to the general population. Although data are limited, risk increases after kidney transplant and with nephrotic syndrome as well. SUMMARY: Rates of kidney disease are increasing rapidly in the population and kidney disease is a risk factor for venous thromboembolism. An improved understanding of mechanisms linking kidney disease with venous thromboembolism will allow further study of best prevention efforts. PMID- 19561508 TI - Management of depression in the last month of life. AB - PURPOSE OF REVIEW: In clinical practice, assessment and adequate treatment of depression in the last month of life remain a difficult task. Although assessment instruments and criteria for depression are available for use in the palliative care population, few have been applied for the last weeks of life. The same is true for clinical management including antidepressants. RECENT FINDINGS: Assessment aids such as Hospital Anxiety and Depression Scale or Hamilton Depression Rating Scale are available and validated for the last days or weeks of life, but its clinical impact remains unclear. The wish to die may symbolize more the acceptance that life comes to an end instead of reflecting a depressive or anxiety disorder. Communication about concrete preparation for death, life completion discussions, expressing religious struggle and giving back a sense of coherence seem to be promising non-pharmacological strategies and may lead to improved social function and better management of physical symptoms. Good news for researchers comes from a recent study highlighting the fact that psychosocial research seems to be of more potential benefit than burden for the patient. SUMMARY: No recent literature can be found on the pharmacological management of depression in the last weeks of life. Few studies demonstrate the usefulness of concrete end of life preparation discussions that underpins the importance and impact of best carer-patient relationship during this last phase of life. A Cochrane protocol on antidepressants in severely physically ill patients has been undertaken recently to highlight this important clinical question, but results may reflect the lack of studies focusing on the last weeks or days of life. PMID- 19561509 TI - The benefits of remote microphone technology for adults with cochlear implants. AB - OBJECTIVE: Cochlear implantation has become a standard practice for adults with severe to profound hearing loss who demonstrate limited benefit from hearing aids. Despite the substantial auditory benefits provided by cochlear implants, many adults experience difficulty understanding speech in noisy environments and in other challenging listening conditions such as television. Remote microphone technology may provide some benefit in these situations; however, little is known about whether these systems are effective in improving speech understanding in difficult acoustic environments for this population. This study was undertaken with adult cochlear implant recipients to assess the potential benefits of remote microphone technology. The objectives were to examine the measurable and perceived benefit of remote microphone devices during television viewing and to assess the benefits of a frequency-modulated system for speech understanding in noise. DESIGN: Fifteen adult unilateral cochlear implant users were fit with remote microphone devices in a clinical environment. The study used a combination of direct measurements and patient perceptions to assess speech understanding with and without remote microphone technology. The direct measures involved a within-subject repeated-measures design. Direct measures of patients' speech understanding during television viewing were collected using their cochlear implant alone and with their implant device coupled to an assistive listening device. Questionnaires were administered to document patients' perceptions of benefits during the television-listening tasks. Speech recognition tests of open set sentences in noise with and without remote microphone technology were also administered. RESULTS: Participants showed improved speech understanding for television listening when using remote microphone devices coupled to their cochlear implant compared with a cochlear implant alone. This benefit was documented both when listening to news and talk show recordings. Questionnaire results also showed statistically significant differences between listening with a cochlear implant alone and listening with a remote microphone device. Participants judged that remote microphone technology provided them with better comprehension, more confidence, and greater ease of listening. Use of a frequency modulated system coupled to a cochlear implant also showed significant improvement over a cochlear implant alone for open-set sentence recognition in +10 and +5 dB signal to noise ratios. CONCLUSIONS: Benefits were measured during remote microphone use in focused-listening situations in a clinical setting, for both television viewing and speech understanding in noise in the audiometric sound suite. The results suggest that adult cochlear implant users should be counseled regarding the potential for enhanced speech understanding in difficult listening environments through the use of remote microphone technology. PMID- 19561510 TI - Smoking as an independent risk factor for myocardial infarction or stroke in type 2 diabetes: a report from the Swedish National Diabetes Register. AB - BACKGROUND: Few earlier studies have analysed smoking as a risk factor for myocardial infarction (MI) or stroke in type 2 diabetic patients. DESIGN AND METHODS: A longitudinal study involved 13 087 female and male patients with type 2 diabetes from the Swedish National Diabetes Register with no previous MI or stroke at baseline, aged 30-74 years, and with data available for all analysed variables, followed up for mean 5.7 years. RESULTS: Adjusted hazard ratios (HRs) for smoking and first-incident fatal/nonfatal MI, stroke and total mortality were 1.7 [95% confidence interval (CI): 1.4-2.0; P<0.001], 1.3 (95% CI: 1.1-1.6; P = 0.006) and 1.8 (95% CI: 1.5-2.2; P<0.001), respectively, by Cox regression analysis, adjusted for age, sex, diabetes duration, hypoglycaemic treatment, haemoglobin A1c, blood pressure, body mass index, microalbuminuria, antihypertensive and lipid-lowering drugs. Adjusted HR was higher for fatal MI, 2.1 (95% CI: 1.7-2.7; P<0.001), than for nonfatal MI, 1.4 (95% CI: 1.2-1.7; P<0.001). The highest HRs were observed in more frequently smoking (22%), middle aged patients (age <60 years) for fatal/nonfatal MI, 2.3 (95% CI: 1.8-3.1; P<0.001) and for total mortality, 2.5 (95% CI: 1.6-3.8, P<0.001), whereas lower HRs were observed in older and less smoking patients. With predicted cessation of smoking in patients aged below 60 years, 24% (95% CI: 15-33%) of cases of fatal/nonfatal MI and 24% (11-37%) of cases of total mortality may have been prevented. CONCLUSION: The risk for MI and total mortality associated with smoking is high in type 2 diabetes, especially in more frequently smoking, middle aged patients, and was higher for MI than for stroke, and also higher for fatal than for nonfatal events. Smoking cessation would strongly affect risk reduction. PMID- 19561511 TI - 5-HTTLPR and STin2 polymorphisms in the serotonin transporter gene and irritable bowel syndrome: effect of bowel habit and sex. AB - BACKGROUND: Conflicting data exist on the association between functional polymorphisms in the serotonin reuptake transporter (SERT) gene (SLC6A4) and irritable bowel syndrome (IBS). This may be partly because of small participant numbers and varying ethnic origin and sex within the cohorts studied. AIM: To reassess the potential association between the SERT polymorphisms 5-HTTLPR and STin2 in both male and female IBS patients with diarrhoea (IBS-D) and constipation (IBS-C) compared with healthy volunteers. METHODS: In this case control study, 196 Caucasian Rome II IBS patients [97 IBS-D (aged 18-66 years; 67 female) and 99 IBS-C (aged 18-65 years; 95 female)] and 92 Caucasian healthy volunteers (aged 18-63 years; 60 female) from the UK had genomic DNA extracted from peripheral blood and the 5-HTTLPR and STin2 polymorphisms genotyped. RESULTS: The frequency of the 5-HTTLPR (ss) genotype was slightly lower in both IBS-D (16.5%) and IBS-C (14.3%) patients compared with controls (23.9%), although not significantly (P or =55%) and reduced EF (<55%) based on the results of the manual analysis. RESULTS: Thirty-two percent of patients had a reduced left ventricular EF of <55%. Volumetric results of the automated inline analysis for EDV (r = 0.96), ESV (r = 0.95), EF (r = 0.89), and MM (r = 0.96) showed high correlation with the results of manual segmentation (all P < 0.001). Head-to-head comparison did not show significant differences between automated and manual evaluation for EDV (153.6 +/- 52.7 mL vs. 149.1 +/- 48.3 mL; P = 0.05), ESV (61.6 +/- 31.0 mL vs. 64.1 +/- 31.7 mL; P = 0.08), and EF (58.0 +/- 11.6% vs. 58.6 +/- 11.6%; P = 0.5). However, differences were significant for MM (150.0 +/- 61.3 g vs. 142.4 +/- 59.0 g; P < 0.01). The standard error was 15.6 (EDV), 9.7 (ESV), 5.0 (EF), and 17.1 (mass). The mean time for manual analysis was 15 minutes. CONCLUSIONS: Unsupervised fully automated segmentation and contouring during image reconstruction enables an accurate evaluation of global systolic cardiac function. PMID- 19561515 TI - Comparison of manual, semi- and fully automated heart segmentation for assessing global left ventricular function in multidetector computed tomography. AB - PURPOSE: To evaluate the reliability of global left ventricular (LV) function and mass measurements with the aid of a semi-automated (Circulation; Siemens, Forchheim, Germany) and a new fully automated software (Philips Research Europe, Aachen, Germany) versus an established manual segmentation method (Argus; Siemens). MATERIAL AND METHODS: Forty-one patients (31 men, 10 women; mean age: 62 +/- 5 years) with known or suspected coronary heart disease underwent contrast enhanced Dual-Source computed tomography of the heart (120 kV, 410 mAs/rotation, collimation 2 x 32 x 0.6 mm, gantry rotation time 0.33 milliseconds). Global LV function measurements of end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume, ejection fraction (EF), and LV mass were each assessed with a manual, a semi- and fully automated method. The latter were compared with the manual contour tracing method, which was considered as standard of reference. Postprocessing time for each method was recorded. For statistical analysis, repeated-measures analysis of variance, post hoc t test, and concordance correlation coefficients were calculated. Bland-Altman plots were generated. RESULTS: In general, ESV and EF assessed with the semi-automated and with the fully automated prototype version agreed well with the manual contour tracing method. The mean ESV (+/-SD) calculated from the manual, the semi-automated, and the fully automated method was 67 +/- 43 mL, 74 +/- 54 mL, and 75 +/- 48 mL, respectively. No statistically significant differences between the methods were found for ESV and EF. In contrast, significant variations (P < 0.05) among the different segmentation methods were shown for EDV, stroke volume, and LV mass. This variation was predominantly due to variation in endocardial delineations among the different techniques. Concordance correlation coefficients demonstrated a better accuracy for the fully automated method than for the semi-automated technique when compared with the manual drawing method. Furthermore, fully automated postprocessing heart segmentation yielded time savings of approximately 80% compared with the manual segmentation tool and 63% compared with the semi automated technique. Mean postprocessing time (+/-SD) for the manual, the semi automated, and the fully automated method was 345 +/- 75 seconds, 192 +/- 58 seconds, and 72 +/- 58 seconds, respectively. CONCLUSION: LV function and mass analyses using semi- or fully automated segmentation algorithms are feasible even if significant differences in EDV assessment are observed. The fully automated method results in better accuracy and time savings when compared with manual and semi-automated data analysis. PMID- 19561516 TI - Three-dimensional imaging of pulmonary veins by a novel steady-state free precession magnetic resonance angiography technique without the use of intravenous contrast agent: initial experience. AB - PURPOSE: To evaluate the feasibility of 3-dimensional (3D) steady-state free precession (SSFP) magnetic resonance angiography (MRA) using nonselective radiofrequency excitation for imaging of pulmonary veins (PVs) without intravenous gadolinium chelate and to correlate the results with conventional contrast-enhanced MRA (CE-MRA). MATERIAL AND METHODS: Forty consecutive patients with history of atrial fibrillation underwent free-breathing respiratory navigator-gated electrocardiogram-triggered SSFP MRA without contrast administration and conventional high-resolution 3D CE-MRA of the thorax at 1.5 T. Two readers assessed both datasets for vascular definition (from 0, not visualized, to 3, excellent definition), artifacts, and ostial diameters. Statistical analysis was performed using Wilcoxon, paired t test, and kappa coefficient. RESULTS: On SSFP MRA, readers 1 and 2 graded 96.4% (160/166) and 97% (161/166) of the segments as having diagnostic visibility and sharpness, respectively (k = 0.82). On CE-MRA datasets, all segments were graded as having diagnostic visibility and sharpness by both readers (k = 0.86). No significant difference existed for visibility and sharpness of pulmonary venous segments between the datasets for each reader (P[r] > 0.05). Reader 1 (2) identified 27 (28) and 35 (32) motion artifacts on SSFP and CE-MRA datasets, respectively. No significant difference was found to exist between ostial diameters on CE-MRA and SSFP datasets (P > 0.05). CONCLUSION: Our study shows that 3D depiction of PVs without intravenous contrast is feasible with nonslice-selective SSFP MRA. This novel MRA technique may be used in certain patients with atrial fibrillation to assess the number and size of PV ostia draining to the left atrium prior to radiofrequency ablation. PMID- 19561517 TI - Regulation of collagen turnover in human skin fibroblasts exposed to a gadolinium based contrast agent. AB - OBJECTIVE: Nephrogenic systemic fibrosis is a clinical syndrome linked with exposure in renal failure patients to gadolinium-based contrast agents (GBCAs) during magnetic resonance imaging. Recently, we demonstrated that GBCA exposure led to increased matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinases-1 (TIMP-1) levels in human skin fibroblasts. The goals of the present work were to assess the relationship between altered MMP-1/TIMP-1 expression and collagen production/deposition, and the intracellular signaling events that lead from GBCA stimulation to altered MMP-1 and TIMP-1 production. MATERIALS AND METHODS: Human dermal fibroblasts were treated with one of the currently used GBCAs (Omniscan). Proliferation was quantified as were levels of MMP-1, TIMP-1, procollagen type I, and collagen type I. Signaling events were concomitantly assessed, and signaling inhibitors were used. RESULTS: Fibroblasts exposed to Omniscan had increases in both MMP-1 and TIMP-1 levels. Omniscan treatment interfered with collagen turnover, leading to increased type I collagen deposition without an increase in type I procollagen production. U0126, an inhibitor of mitogen-activated protein kinase signaling, and LY294002, a phosphatidylinositol-3 kinase inhibitor, reduced MMP-1 levels. U0126 also reduced TIMP-1 levels, but LY294002 increased TIMP-1. CONCLUSION: These data provide evidence for complex regulation of collagen deposition in Omniscan-treated skin. They suggest that the major effect of Omniscan exposure is on an enzyme/inhibitor system that regulates collagen breakdown rather than on collagen production, per se. PMID- 19561518 TI - Improving outcomes in state AIDS drug assistance programs. AB - BACKGROUND: State AIDS Drug Assistance Programs (ADAPs) provide antiretroviral medications to patients with no access to medications. Resource constraints limit the ability of many ADAPs to meet demand for services. OBJECTIVE: To determine ADAP eligibility criteria that minimize morbidity and mortality and contain costs. METHODS: We used Discrete Event Simulation to model the progression of HIV infected patients and track the utilization of an ADAP. Outcomes included 5-year mortality and incidence of first opportunistic infection or death and time to starting antiretroviral therapy (ART). We compared expected outcomes for 2 policies: (1) first-come first-served (FCFS) eligibility for all with CD4 count or = 65 years) for baseline characteristics, changes in selected measures during CR, and the proportion of patients at secondary prevention treatment goals before and after CR. Subanalyses were conducted between "young-old" (65-74 years) and "old-old" (> or = 75 years) patients. RESULTS: At baseline, older patients had lower body mass indexes, better lipid profiles, lower hemoglobin A1c levels (when diabetes was present), and lower Beck Depression Inventory scores (all P < .05) but had higher blood pressures, more comorbidities, and poorer functional capacity as demonstrated by shorter 6-minute walk distance (all P < .05). At CR completion, improvement (P < .05) was achieved among younger patients for all measures except for high-density lipoprotein cholesterol and among older patients for all measures except for diastolic blood pressure and high-density lipoprotein cholesterol. Similar improvements from baseline to CR completion were evident among the "old-old." While the degree of improvement in individual outcomes varied by age, age group was not a significant predictor of achieving secondary prevention goals at CR completion. CONCLUSIONS: Older patients with CHD entered CR with less adverse risk factors but had a higher comorbidity burden than did younger patients. Both groups exhibited significant improvements by CR completion, and these improvements extended to the oldest patients. PMID- 19561519 TI - A randomized comparative trial of continued zidovudine/lamivudine or replacement with tenofovir disoproxil fumarate/emtricitabine in efavirenz-treated HIV-1 infected individuals. AB - BACKGROUND: Long-term antiretroviral therapy dramatically reduces HIV-related morbidity and mortality but is also associated with metabolic and morphological changes and requires high levels of adherence. METHODS: A randomized, 48-week, open-label, comparative study of continuation of twice-daily zidovudine/lamivudine or replacement with once-daily tenofovir disoproxil fumarate/emtricitabine in individuals on successful efavirenz-based antiretroviral therapy. Limb fat mass was assessed by dual x-ray absorptiometry in a subset of participants through week 48. RESULTS: Two hundred thirty-four individuals were randomized and treated (117 each to continue or switch) with 206 subjects completing 48 weeks of study. Five percent subjects in the continue group and 3% subjects in the switch group discontinued due to adverse events. By intent-to-treat, missing = failure analysis, no differences in virological efficacy were observed at any time point (week 48 <50 copies/mL continue 85%, switch 88%). At week 24, switching was associated with significant increases in hemoglobin (mean difference 0.37 g/dL, 95% confidence interval: 0.15 to 0.58 g/dL, P < 0.001) and significant declines in total cholesterol and triglycerides. In the dual x-ray absorptiometry substudy (n = 100), fat was preserved or increased in the switch group but declined in the continue group (mean difference 448 g, 95% confidence interval: 57 to 839 g, P = 0.025). Individuals with longer exposure to zidovudine and lower baseline limb fat experienced less limb fat increase after switching. No differences in renal adverse events were observed between groups. CONCLUSIONS: Switching from zidovudine/lamivudine to tenofovir disoproxil fumarate/emtricitabine in persons on efavirenz therapy maintains virological control, establishes a once-daily regimen, results in improvements in hemoglobin and key lipid parameters, and preserves and restores limb fat relative to continuation of zidovudine/lamivudine. PMID- 19561521 TI - Resistance exercise training improves heart function and physical fitness in stable patients with heart failure. AB - PURPOSE: This study determined the effect of a structured isotonic strength training (ST) program on left ventricular (LV) function (ejection fraction, stroke volume, and end-diastolic and end-systolic volumes) and physical fitness (6-minute walk test, upper body strength, lower body strength, and body composition) in patients with New York Heart Association class II and III heart failure. METHODS: Sixteen patients were randomized into 2 groups, ST and usual care. The ST group (10 patients) performed 24 ST exercise sessions (3 per week, 8 weeks), while the usual care (6 patients) group followed routine medical care. The structured isotonic ST program involved 12 different exercises on circuit weight machines. LV function (3D echocardiography) and physical fitness were assessed at baseline and 8 weeks. RESULTS: Modest improvements (P < .05) in resting ejection fraction (0.32-0.37) and stroke volume (46 to 53 mL/beat), as well as in muscular strength and 6-minute walk distance, were found after training. CONCLUSIONS: A short-term structured isotonic ST program appears to improve selected measures of resting LV function and fitness in patients with mild congestive heart failure. Additional studies utilizing larger numbers of subjects, including women, are needed. PMID- 19561523 TI - Pulmonary rehabilitation after acute exacerbation of chronic obstructive pulmonary disease in patients who previously completed a pulmonary rehabilitation program. AB - PURPOSE: Acute exacerbations of chronic obstructive pulmonary disease (AECOPD) impair health-related quality of life (HRQL). We evaluated the effect of an abbreviated repeat pulmonary rehabilitation (PR) program on HRQL after an AECOPD. METHODS: Patients who had completed PR were followed for up to 12 months to identify an AECOPD and then placed in randomized groups to receive a 3-week repeat-PR intervention or usual care. Measures of HRQL (Chronic Respiratory Disease Questionnaire, CRQ) and functional exercise capacity (6-minute walk distance, 6MWD) were collected at 2 (T(1)), 5 (T(2)), and 12 weeks (T(3)) post AECOPD. The repeat-PR program was undertaken between T(1) and T(2). Between-group differences were examined using repeated- measures analysis of variance or covariance. RESULTS: Of the 60 patients (30 men, age 69+/-8 years, forced expiratory volume in 1 second 0.86+/-0.40 L, 6MWD 367+/-99 m) followed, 41 experienced an AECOPD 14 +/- 11 weeks after completion of the initial PR program and 33 completed the study. Of these, 16 and 17 were randomized to the intervention and control groups, respectively. No between-group differences were demonstrated at T(2) or T(3). With the exclusion of 5 subjects who experienced a second AECOPD between T(1) and T(3), the participants in the intervention group demonstrated greater reduction in dyspnea when compared to those in the control group at T(3) (0.8+/-1.6 vs -0.4+/-1.3 points per item, P = .04). CONCLUSIONS: The reduction in dyspnea in those who did not experience a second AECOPD provides preliminary evidence for the role of repeat programs. The application of repeat PR should be refined in larger trials. PMID- 19561522 TI - Relationship of resting B-type natriuretic peptide level to cardiac work and total physical work capacity in heart failure patients. AB - PURPOSE: Plasma B-type natriuretic peptide (BNP) levels obtained at rest have been previously shown to be correlated with the global functional capacity measures of peak oxygen uptake (V(O(2peak))) and the minute ventilation/carbon dioxide (VE/V(O(2))) slope. The purpose of this study was to assess the relationship of the plasma BNP level to the rate-pressure product (RPP) as an indicator of central or cardiac work capacity. METHODS: Twenty-two subjects (12 men), mean age 57 +/- 12 years, diagnosed with heart failure (8 ischemic/14 nonischemic) were recruited. All subjects were stable on optimal medical therapy for at least 1 month. Blood samples for BNP level analysis were obtained at rest. Subjects underwent a symptom-limited treadmill exercise test using a ramping protocol while V(O(2)), heart rate (HR), and blood pressure (BP) were monitored. Correlation analyses were conducted to assess the relationship of BNP level to RPP level, V(O(2peak), VE/V(O(2)) slope, end-tidal CO(2) pressure (P(ET)CO(2)), and left ventricular ejection fraction (LVEF). RESULTS: Resting BNP levels were significantly correlated with RPP levels (r = -0.69). The BNP level and the RPP level were correlated with V(O(2peak)) (r = -0.63 and r = 0.66, respectively) and VE/V(O(2)) slope (r = 0.53 and r = -0.54, respectively). The RPP level but not the BNP level was correlated with P(ET)CO(2) (r = 0.57). Neither BNP nor RPP levels were well correlated with LVEF (r = -0.26 and r = 0.14, respectively). DISCUSSION: The results of this study suggest that resting plasma BNP level may be a useful clinical measure for evaluating both global functional capacity and myocardial specific work capacity in individuals with heart failure. PMID- 19561524 TI - Home-based pulmonary rehabilitation in very severe COPD: is it safe and useful? AB - OBJECTIVE: To determine the safety and usefulness of a home-based pulmonary rehabilitation program for patients with very severe chronic obstructive pulmonary disease (COPD) who receive long-term treatment with oxygen. METHODS: A randomized prospective study that included 50 patients with very severe COPD undergoing long-term treatment with oxygen. At the start of the program and after 12 months, the following evaluations were performed: pulmonary function tests, submaximal exercise capability, and quality of life. The rehabilitation program lasted for 1 year, with periodic home visits by a physiotherapist and included respiratory reeducation training of the respiratory muscles and muscular training. RESULTS: Forty-two patients completed the study (27 in the rehabilitation group and 15 in the control group). There were no initial differences between the 2 groups. No complications arising from the performance of the exercises were observed. The rehabilitation group showed a clinically significant increase in the distance covered in the 6-minute walk test (313 +/- 72 m vs 392 +/- 82 m, P = .0001) and a clinically significant reduction in the St George's Respiratory Questionnaire score (55.3 +/- 15.0 vs 40.5 +/- 13.8, P = .0001). No significant changes were found in any of the parameters studied in the control group. CONCLUSION: A home-based pulmonary rehabilitation program for patients with very severe COPD under long-term oxygen treatment is safe and useful, as it achieves an improvement in exercise tolerance, reduces dyspnea after effort, and improves quality of life without causing any complication arising from the performance of the exercises. PMID- 19561526 TI - Novel anticoagulants in clinical development: focus on factor Xa and direct thrombin inhibitors. AB - Vitamin K antagonists are the mainstay in the prevention and treatment of thromboembolic diseases. Although effective under optimal conditions, several drawbacks are imminent to the long-term application of these drugs due to their narrow therapeutic window, interactions with other drugs as well as the need for regular monitoring and the risk of a recurrent event versus the risk of bleeding. To overcome these downsides, novel anticoagulants are being developed; in contrast to vitamin K antagonists, these novel agents specifically and selectively block central elements of the coagulation cascade. Several clinical trials have demonstrated the efficacy and safety of selective FXa inhibitors (such as fondaparinux, rivaroxaban, apixaban) and direct thrombin inhibitors (such as lepirudin, bivalirudin, dabigatran etexilate) in the treatment of typical indications for conventional vitamin K antagonists, in particular, the prevention and treatment of venous thromboembolism. This review summarizes the results and designs of recently published and ongoing clinical trials of novel anticoagulants. PMID- 19561525 TI - Does stress reduction change the levels of cortisol secretion in patients with coronary artery disease? AB - PURPOSE: It has been shown that behavioral therapy has effects on stress behavior in patients with coronary artery disease. Salivary cortisol measurements are widely used to assess psychological stress, stress reactivity, or both. The aim of this study was to investigate whether improved stress behavior in type A patients with coronary artery disease involved changes in cortisol secretion pattern. METHODS: Twenty-four male patients were identified as type A individuals and completed a 12-month cognitive-behavioral stress management program. Stress behavior was evaluated by using a validated questionnaire. Morning and evening salivary cortisol levels were measured over 3 consecutive days at baseline and after 12 months. RESULTS: Although the patients showed a significant improvement in psychosocial well-being after 12 months, their basal cortisol levels or diurnal rhythm of cortisol did not change. There was no correlation between stress score and cortisol levels. CONCLUSIONS: The value of salivary cortisol as both a stress marker and a new cardiovascular risk factor has been discussed but the data from this small pilot study raise the question of its utility as a stress marker in cardiac rehabilitation. PMID- 19561527 TI - 5-Fluorouracil-induced coronary vasospasm. AB - Cardiotoxicity due to 5-fluorouracil involves rare and life-threatening cardiotoxic events occurring in less than 1% of the patients. We describe a case of coronary vasospasm due to parenteral 5-fluorouracil, which did not recur with oral capecitabine. We also give a brief review of the cardiotoxicity of 5 fluorouracil, its treatment and prevention. PMID- 19561528 TI - Undiagnosed maternal celiac disease in pregnancy and an increased risk of fetal growth restriction. PMID- 19561529 TI - Stomach cancer risk in gastric cancer relatives: interaction between Helicobacter pylori infection and family history of gastric cancer for the risk of stomach cancer. AB - GOALS: To identify the risk of gastric cancer in first-degree relatives of gastric cancer patients, and to determine if there is an interaction between Helicobacter pylori (H. pylori) infection and family history of gastric cancer in gastric carcinogenesis. BACKGROUND: It is unclear to what degree a family history of gastric cancer is associated with stomach cancer risk in Korea. STUDY: From May 2003 to July 2008, 428 gastric cancer patients and 368 controls were included in the analyses. Logistic regression models including age, sex, family history of gastric cancer, residency during childhood, smoking, monthly income, spicy food diet and H. pylori status were evaluated to estimate the odds ratios (ORs) of developing gastric cancer. RESULTS: Adjusted OR for gastric cancer increased 3 fold for subjects reporting first-degree relatives with gastric cancer [OR 2.85, 95% confidence interval (CI): 1.83-4.46]. The association was strong in the 40 to 59 years age group (OR 4.00, 95% CI: 2.06-7.76), and became weaker in subjects older than 60 years of age (OR 1.81, 95% CI: 0.95-3.46). Compared with the uninfected subjects without a family history, subjects with both a family history and H. pylori infection had a 5-fold increased risk (OR 5.32, 95% CI: 2.76 10.25). CONCLUSIONS: After adjusting for environmental factors and H. pylori infection, a family history of gastric cancer remained independently associated with gastric cancer. The interaction between H. pylori infection and family history of gastric cancer might be a rationale for H. pylori eradication in the gastric cancer relatives as a strategy to prevent gastric cancer. PMID- 19561530 TI - Efficacy of 5-ASA in the treatment of colonic diverticular disease. AB - OBJECTIVE: To assess the therapeutic efficacy of 5-aminosalicylic acid (5-ASA) in patients with colonic diverticular disease performing a systematic review of the literature. DATA SOURCES: Cochrane Trial Register (until Issue 4, 2008), Medical Literature Analysis and Retrieval System Online (1966 to October 1, 2008), Excerpta Medica Database (1980 to October 1, 2008), and abstracts from the major US, European, and Asian gastroenterology conferences. Expert opinions sought and reference lists of identified studies and any relevant published reviews checked. STUDY SELECTION: Randomized controlled trial (RCT) or a controlled clinical trial with a parallel group design using 5-ASA as 1 treatment arm. RESULTS: Six RCTs enrolling 818 patients were found: 3 were performed in patients with uncomplicated diverticulitis, and the remaining in patients with symptomatic uncomplicated diverticular disease. The results of these studies showed that patients treated with 5-ASA had significantly better outcomes and that also mesalazine scheduled daily was superior to cyclic administration to prevent relapse of diverticular disease. However, several of these studies have not included an endoscopy at the start of the study and when patients had recurrences. CONCLUSIONS: From this systematic review, it seems that 5-ASAs may have a role in patients with colonic diverticular disease. However, high-quality well-designed RCTs are necessary to confirm these initial observations. PMID- 19561531 TI - The prevalence of colorectal neoplasia in patients with gastric cancer: a Korean Association for the Study of Intestinal Disease (KASID) Study. AB - GOALS: The goal of this study was to determine the prevalence of colorectal neoplasia, using colonoscopy surveillance, in a cohort of patients with gastric cancers. BACKGROUND: The association between gastric cancer and colorectal cancer has been conflicting. STUDY: A total of 543 patients (males, 362; females, 181) with gastric cancer were matched with 2 persons from the population without a diagnosis of gastric cancer as confirmed by endoscopy according to age (+/-2 y), sex, date of colonoscopy examination (+/-2 wk), and endoscopist. Main Outcome was the prevalence of colorectal neoplasia diagnosed by colonoscopy. RESULTS: A significantly higher colorectal cancer prevalence was found in the gastric cancer group, that is, 19 of 543 (3.5%) versus 14 of 1086 (1.3%; P<0.001). The odds of developing colorectal cancer were higher in the presence of gastric cancer (odds ratios, 3.46; 95% confidence interval: 1.51-7.91). Four of the 119 (3.4%) gastric cancer patients below 50 years of age had colorectal cancer in contrast with no cases in the matched controls. The prevalence of colorectal adenoma was higher in the gastric cancer group, with a prevalence of 215 in 543 (39.6%) versus 311 in 1086 (28.6%; P<0.001). The risk of adenoma was also greater among gastric cancer patients (odds ratios, 1.76; 95% confidence interval: 1.34-2.25). CONCLUSIONS: Our data reveal a higher prevalence and risk of colorectal cancer in patients diagnosed with gastric cancer, particularly in patients below 50 years of age. Additional studies are needed to explore the geographical differences in the association between gastric cancer and colon cancer. PMID- 19561532 TI - Vaccination of renal cell cancer patients with modified vaccinia Ankara delivering the tumor antigen 5T4 (TroVax) alone or administered in combination with interferon-alpha (IFN-alpha): a phase 2 trial. AB - Attenuated vaccinia virus, modified vaccinia Ankara (MVA) has been engineered to deliver the tumor antigen 5T4 (TroVax). MVA-5T4 has been evaluated in an open label phase 2 trial in metastatic renal cell cancer patients in which the vaccine was administered alone or in combination with interferon-alpha-2b (IFN-alpha). The safety, immunologic, and clinical efficacy of MVA-5T4 with or without IFN alpha was determined. Twenty-eight patients with metastatic renal cell cancer were treated with MVA-5T4 alone (13) or plus IFN-alpha (15). The 5T4-specific cellular and humoral responses were monitored throughout the study. Clinical responses were assessed by measuring changes in tumor burden by computed tomography or magnetic resonance imaging scan. MVA-5T4 was well tolerated with no serious adverse event attributed to vaccination. Of 23 intent-to-treat patients tested for immune responses postvaccination, 22 (96%) mounted 5T4-specific antibody and/or cellular responses. One patient treated with MVA-5T4 plus IFN alpha showed a partial response for >7 months, whereas an additional 14 patients (7 receiving MVA-5T4 plus IFN and 7 receiving MVA-5T4 alone) showed periods of disease stabilization ranging from 1.73 to 9.60 months. Median progression free survival and overall survival for all intent-to-treat patients was 3.8 months (range: 1 to 11.47 mo) and 12.1 months (range: 1 to 27 mo), respectively. MVA-5T4 administered alone or in combination with IFN-alpha was well tolerated in all patients. Despite the high frequency of 5T4-specific immune responses, it is not possible to conclude that patients are receiving clinical benefit. The results are encouraging and warrant further investigation. PMID- 19561534 TI - A novel mouse model for evaluation and prediction of HLA-A2-restricted CEA cancer vaccine responses. AB - Human leukocyte antigen (HLA)-A2.1 transgenic mice (HHD) represent a valuable model to study and predict the immunogenicity of vaccines against pathogens. However, HHD mice are unsuitable for in vivo studies of cancer vaccines against human tumor-associated antigens because they lack T-cell tolerance that is key to define the potency of the treatment. In this study, we developed HHD/carcinoembryonic antigen P(CEA) hybrid mice by breeding transgenic mice homozygous for CEA with HHD. These mice express human CEA, present epitopes solely through HLA-A2.1 molecules and constitute a unique in vivo animal model to study HLA-A2.1-restricted immune response of a human CEA-based vaccine. Owing to the immune tolerance, HHD/CEA mice show a limited immune response and expansion of a different and restricted T-cell receptor repertoire after antigen-specific stimulation. Our data show that genetic vectors expressing CEA and peptide-based vaccines are able to efficiently break immune tolerance against CEA and to elicit strong immune response against HLA-A2.1-restricted CEA epitopes. Most importantly, efficient lysis of human CEA+/HLA-A2.1+ tumor cells was observed and significant protection against HHD/CEA tumor cells was achieved in HHD/CEA vaccinated mice. Hence, HHD/CEA provides a relevant model for the evaluation of the potential efficacy of human CEA-based vaccines. PMID- 19561533 TI - HSCT recipients have specific tolerance to MSC but not to the MSC donor. AB - Multipotent mesenchymal stromal cells (MSC) are increasingly used to treat refractory graft-versus-host-disease and other complications after hematopoietic stem cell transplantation (HSCT). We evaluated immunogenicity of HLA-mismatched MSC infused posttransplant to HSCT recipients. Recipient lymphocyte response to MSC and peripheral blood lymphocytes (PBL) from the MSC or third party donors was measured before and after infusion. In vitro primary and rechallenge lymphocyte responses of healthy individuals to MSC and to PBL from the MSC donor were similarly studied. HSCT recipients given MSC responded to third party allostimuli, but showed no response to infused MSC before and upto 6 months after infusion, whereas maintaining an alloresponse to the MSC donor. This indicates immune unresponsiveness restricted to MSC, as the HSCT recipient was not tolerized to the MSC donor. In vitro, we confirmed that MSC failed to prime responder lymphocytes to rechallenge with PBL from the MSC donor, and lymphocytes primed with MSC donor and rechallenged with MSC only showed weak responses at high stimulator-responder ratios. Although MSC up-regulated lymphocyte gene expression of CD25, IFN-gamma, FoxP3, CTLA-4, and IL-10, they failed in both unprimed and primed responders to induce CD25+ (activated) or CD57+ (effector) CD4+ or CD8+ T-lymphocyte subsets and only inconsistently induced FoxP3+ regulatory T-lymphocytes. These results show for the first time that infused MSC are only weakly immunogenic in humans and validate the clinical use of MSC from HLA-mismatched donors. PMID- 19561535 TI - Activated T-cell-mediated immunotherapy with a chimeric receptor against CD38 in B-cell non-Hodgkin lymphoma. AB - T-cell-mediated immunotherapy with a chimeric antigen receptor (CAR) is expected to become a powerful treatment for cancer. CD38, highly expressed in B-cell non Hodgkin lymphoma (B-NHL) cells, is an attractive target in immunotherapy for B NHL. We retrovirally transduced a T-cell line, Hut78, expressing little CD38, with an anti-CD38-CAR. Hut78 cells with the anti-CD38-CAR were cocultured with B NHL cell lines bearing CD38 and also B-NHL cells from patients. Four days later most of the lymphoma cells were killed (the level of cytotoxicity was >95%). By contrast, there was undetectable cytotoxicity against CD38-negative cell lines. Then, we introduced the anti-CD38-CAR into human peripheral T cells. However, the recovery of viable cells was very low, presumably because of an autolytic reaction caused by the association of the anti-CD38-CAR with CD38 on the cell surface. The addition of an anti-CD38 antibody increased the yield of viable transduced T cell probably by blocking the autolytic reaction. We cocultured human peripheral T cells bearing anti-CD38-CAR with B-NHL cells. The median specific cytotoxicity was greater than 90%. These cells were injected 4 times into NOD/SCID mice, which were inoculated with B-NHL cells luciferase. Luciferase activity was not detectable even 30 days after the inoculation in 5 of 6 mice injected. By contrast, it increased in all of the mice injected with the mock vector-transduced T cell. In conclusion, T cell with the anti-CD38-CAR showed powerful cytotoxicity against B-NHL cells in vitro and in vivo. These findings may provide an important clue for improving the methodology of T-cell-mediated immunotherapy. PMID- 19561537 TI - Inhibition of tumor growth by targeted toxins in mice is dramatically improved by saponinum album in a synergistic way. AB - The application of targeted toxins in cancer therapy remains a challenge due to the severe side effects as a consequence of the high systemic doses required. Here, we describe the combined application of a glycosylated triterpenoid (Spn) and epidermal growth factor receptor (EGFR)-targeted chimeric toxins (SA2E). The cytotoxicity of SA2E on murine TSA tumor cells transfected with human EGFR was enhanced 20,000-fold by low nonpermeabilizing Spn concentrations in a synergistic manner. Subcutaneous application of Spn and SA2E in BALB/c mice bearing a solid TSA cells transfected with epidermal growth factor receptor tumor resulted in 94% tumor volume reduction with a 50-fold lower chimeric toxin concentration compared with pure SA2E treatment. Side effects as monitored by observable complications, body weight, blood parameters; histologic analyses and antibody responses were only moderate and usually reversible. PMID- 19561538 TI - Effect of yeast-derived beta-glucan in conjunction with bevacizumab for the treatment of human lung adenocarcinoma in subcutaneous and orthotopic xenograft models. AB - Human lung cancer is the leading cause of cancer death worldwide. Bevacizumab, a monoclonal antibody against vascular endothelial growth factor (VEGF), in combination with chemotherapy showed significant therapeutic efficacy in human lung cancer patients. However, increased adverse effects limit its clinical utilization. Previous studies demonstrated that polysaccharide beta-glucan significantly augments antitumor monoclonal antibody-mediated efficacy via stimulation of the innate effector neutrophil complement receptor 3. Here, we explored combined beta-glucan with bevacizumab therapy for human lung cancer using murine xenograft models. To that end, human lung adenocarcinomas were screened for membrane-bound VEGF expression. Both subcutaneous and orthotopic lung cancer xenograft models were used to evaluate the combination therapy. We found that PC14PE6 adenocarcinoma cells express membrane-bound VEGF both in vitro and in vivo. Bevacizumab bound to surface VEGF on PC14PE6 cells and activated complement. In the subcutaneous PC14PE6 tumor model, beta-glucan plus bevacizumab showed augmented efficacy in terms of tumor progression and long-term survival compared with bevacizumab-treated alone. These effects were accompanied with massive complement deposition and neutrophil infiltration within tumors. However, this effect was not observed in surface-bound VEGF-negative human lung tumors. Therapeutic efficacy of beta-glucan with bevacizumab was further demonstrated in an orthotopic lung cancer model. Thus, our data suggest that beta-glucan enhances bevacizumab-mediated efficacy and may provide therapeutic benefits for lung cancers with membrane-bound VEGF expression. PMID- 19561536 TI - Genetic modification of T cells with IL-21 enhances antigen presentation and generation of central memory tumor-specific cytotoxic T-lymphocytes. AB - An optimized antigen-presenting cell for tumor immunotherapy should produce a robust antigen specific cytotoxic T lymphocytes (CTL) response to tumor associated antigens, which can persist in vivo and expand on antigen reencounter. Interleukin (IL)-21 synergizes with other gamma-chain cytokines to enhance the frequency and cytotoxicity of antigen-specific CTL. As T cells themselves may serve as effective antigen-presenting cells (T antigen-presenting cells; TAPC) and may be useful in vivo as cellular vaccines, we examined whether CD8(+) T cells genetically modified to produce IL-21 could induce immune responses to tumor associated antigen peptides in healthy human leukocyte antigen-A2(+) donors. We found that IL-21 modified TAPC enhanced both the proliferation and survival of MART-1 specific CD8(+) T cells, which were enriched by >8-fold over cultures with control nontransgenic TAPC. MART-1-specific CTL produced interferon gamma in response to cognate peptide antigen and killed primary tumor cells expressing MART-1 in a major histocompatibility complex restricted manner. IL-21 modified TAPC similarly enhanced generation of functional CTL against melanoma antigen gp100 and the B-cell chronic lymphocytic leukemia associated RHAMM antigen. Antigen-specific CTL generated using IL-21 gene-modified TAPC had a central memory phenotype characterized by CD45RA(-), CD44(high), CD27(high), CD28(high), CD62L(high), and IL-7 receptor-alpha(high), contrasting with the terminal effector phenotype of CTL generated in the absence of IL-21. Thus, TAPC stimulation in the presences of IL-21 enhances proliferation of tumor antigen specific T cells and favors induction of a central memory phenotype, which may improve proliferation, survival, and efficacy of T-cell based therapies for the treatment of cancer. PMID- 19561539 TI - Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor. AB - T cells can be engineered to express the genes of chimeric antigen receptors (CARs) that recognize tumor-associated antigens. We constructed and compared 2 CARs that contained a single chain variable region moiety that recognized CD19. One CAR contained the signaling moiety of the 4-1BB molecule and the other did not. We selected the CAR that did not contain the 4-1BB moiety for further preclinical development. We demonstrated that gammaretroviruses encoding this receptor could transduce human T cells. Anti-CD19-CAR-transduced CD8+ and CD4+ T cells produced interferon-gamma and interleukin-2 specifically in response to CD19+ target cells. The transduced T cells specifically killed primary chronic lymphocytic leukemia (CLL) cells. We transduced T cells from CLL patients that had been previously treated with chemotherapy. We induced these T cells to proliferate sufficiently to provide enough cells for clinical adoptive T cell transfer with a protocol consisting of an initial stimulation with an anti-CD3 monoclonal antibody (OKT3) before transduction followed by a second OKT3 stimulation 7 days after transduction. This protocol was successfully adapted for use in CLL patients with high peripheral blood leukemia cell counts by depleting CD19+ cells before the initial OKT3 stimulation. In preparation for a clinical trial that will enroll patients with advanced B cell malignancies, we generated a producer cell clone that produces retroviruses encoding the anti-CD19 CAR, and we produced sufficient retroviral supernatant for the proposed clinical trial under good manufacturing practice conditions. PMID- 19561541 TI - Nodular collagenous gastritis. PMID- 19561542 TI - Infliximab therapy in children with concurrent perianal Crohn disease: observations from REACH. AB - OBJECTIVE: Post hoc analyses evaluated the effect of infliximab upon concurrent perianal Crohn disease (CD) in a subpopulation of 31 patients from REACH, a randomized trial of 112 children with moderately to severely active luminal CD. MATERIALS AND METHODS: The Pediatric Crohn Disease Activity Index perirectal subscore was used to assess perianal symptom activity and therapeutic response. Patients with no symptoms or asymptomatic tags received a score of 0; those with "1-2 indolent fistula, scant drainage, no tenderness" received a score of 5; and those with "active fistula, drainage, tenderness or abscess" received a score of 10. Initial perirectal subscores of 10 or 5 decreasing to 0 were considered complete response. Subscores of 10 decreasing to 5 were considered partial response. All patients were followed for efficacy and safety through week 54. RESULTS: Twenty-two patients with baseline perianal disease were randomized at week 10 following a 3-dose infliximab induction regimen. At week 2, 40.9% (9/22) of patients with signs and symptoms of perianal disease at baseline attained response (4 partial and 5 complete). At week 54, 72.7% (16/22) of patients with signs and symptoms of perianal disease attained response (1 partial and 15 complete). Nine patients developed perianal signs and symptoms during treatment; 7 had complete response and 2 had no response at week 54. The incidence of adverse events for patients with perianal symptoms at baseline and for those in the overall REACH population was similar (95.7% vs 94.6%). CONCLUSIONS: Infliximab rapidly reduced concurrent perianal disease signs and symptoms in this REACH cohort. PMID- 19561543 TI - Autoimmune hepatitis. AB - Autoimmune hepatitis is characterized by inflammatory liver histology, circulating nonorgan-specific autoantibodies, and increased levels of immunoglobulin G, in the absence of a known etiology. Two types of juvenile autoimmune hepatitis (AIH) are recognized according to seropositivity for smooth muscle and/or anti-nuclear antibody (AIH type 1) or liver kidney microsomal antibody (AIH type 2). There is a female predominance in both. AIH type 2 presents more acutely, at a younger age and commonly with immunoglobulin A deficiency, whereas duration of symptoms before diagnosis, clinical signs, family history of autoimmunity, presence of associated autoimmune disorders, response to treatment, and long-term prognosis are similar in the 2 groups. Immunosuppressive treatment with steroids and azathioprine, which should be instituted promptly to avoid progression to cirrhosis, induces remission in 80% of cases. Relapses are common, often due to nonadherence. Drugs effective in refractory cases include cyclosporine and mycophenolate mofetil. Long-term treatment is usually required, with only some 20% of AIH type 1 patients able to discontinue therapy successfully. In childhood, sclerosing cholangitis with strong autoimmune features, including interface hepatitis and serological features identical to AIH type 1, is as prevalent as AIH, but it affects boys and girls equally. Differential diagnosis relies on cholangiographic studies. In autoimmune sclerosing cholangitis liver parenchymal damage responds satisfactorily to immunosuppressive treatment, whereas bile duct disease tends to progress. In this article we review the state of the art of diagnosis, monitoring, and treatment for children with AIH. PMID- 19561544 TI - A biodegradable esophageal stent in the treatment of a corrosive esophageal stenosis in a child. PMID- 19561540 TI - High-avidity autoreactive CD4+ T cells induce host CTL, overcome T(regs) and mediate tumor destruction. AB - Despite progress made over the past 25 years, existing immunotherapies have limited clinical effectiveness in patients with cancer. Immune tolerance consistently blunts the generated immune response, and the largely solitary focus on CD8+ T cell immunity has proven ineffective in the absence of CD4+ T cell help. To address these twin-tier deficiencies, we developed a translational model of melanoma immunotherapy focused on the exploitation of high-avidity CD4+ T cells that become generated in germline antigen-deficient mice. We had previously identified a tyrosinase-related protein-1 specific HLA-DRB1*0401-restricted epitope. Using this epitope in conjunction with a newly described tyrosinase related protein-1 germline-knockout, we demonstrate that endogenous tyrosinase related protein-1 expression alters the functionality of the autoreactive T cell repertoire. More importantly, we show, by using major histocompatibility complex mismatched combinations, that CD4+ T cells derived from the self-antigen deficient host indirectly triggers the eradication of established B16 lung metastases. We demonstrate that the treatment effect is mediated entirely by endogenous CD8+ T cells and is not affected by the depletion of host regulatory T cells. These findings suggest that high-avidity CD4+ T cells can overcome endogenous conditions and mediate their antitumor effects exclusively through the elicitation of CD8+ T cell immunity. PMID- 19561545 TI - Partial external biliary diversion in children with progressive familial intrahepatic cholestasis and Alagille disease. AB - BACKGROUND: Partial external biliary diversion (PEBD) is a promising treatment for children with progressive familial intrahepatic cholestasis (PFIC) and Alagille disease. Little is known about long-term outcomes. PATIENTS AND METHODS: A retrospective chart review of all patients undergoing PEBD in the University Medical Centre of Groningen (UMCG). RESULTS: Between 2000 and 2005, PEBD was performed on 14 children with severe pruritus (PFIC 11, mean age 5.3 +/- 4.4 years; Alagille 3, mean age 7.4 +/- 4.2 years). Stature was <-2 standard deviation score (SDS) in 50%. Median preoperative serum bile salt concentration was 318 micromol/L (range 23-527 micromol/L). Twenty-nine percent had severe liver fibrosis and 71% had mild or moderate fibrosis. Median follow-up was 3.1 years (range 2.0-5.7 years). One patient (7%) underwent a liver transplantation at 3.2 years post-PEBD. Two years postoperatively, 50% were without pruritus and 21% had mild pruritus. In 29%, pruritus had not diminished; 3 of them had severe fibrosis preoperatively. In patients with mild or moderate fibrosis, PEBD decreased serum bile salts (105 micromol/L [range 8-269 micromol/L] 2 years postoperatively). Bile salts did not decrease in the patients with severe fibrosis. Two years after PEBD, 27% had a stature below -2 SDS. CONCLUSIONS: At median follow-up of 3.1 years after PEBD, pruritus has been relieved in 75%. Bile salts level and growth are improved in most patients. Longer follow-up is needed to determine whether PEBD can postpone or avoid the demand for liver transplantation. PMID- 19561546 TI - Perianal disease as the initial presentation of autoimmune neutropenia. PMID- 19561547 TI - Risk of capsule endoscope retention in pediatric patients: a large single-center experience and review of the literature. AB - OBJECTIVES: Capsule retention is a potential complication of capsule endoscopy (CE). The aims of our study were to determine the incidence of capsule retention in pediatric patients undergoing CE and to identify potential risk factors for capsule retention. MATERIALS AND METHODS: We performed an institutional review board-approved retrospective chart review of pediatric patients undergoing CE studies at a single center. Data collected included patient age, sex, prior diagnosis of inflammatory bowel disease (IBD), CE indication, prior small bowel series results, study result, and complications. RESULTS: Two hundred seven CE procedures were performed in pediatric patients during the study period. Capsule retention occurred in 3 (1.4%) of the 207 studies. All 3 patients had known Crohn disease (CD). The risk of capsule retention in pediatric patients with known IBD was 5.2% (3/58). The risk of capsule retention for patients with suspected IBD and all other indications was 0%. If small bowel disease was identified on upper gastrointestinal series in patients with known CD, then the risk of capsule retention was 37.5% (3/8). Only 7 patients with known IBD had a body mass index (BMI) below the 5th percentile. Of these 7 patients, 3 (43%) had capsule retention. CONCLUSIONS: Red flags for potential CE retention identified in our study include known IBD (5.2% retention risk), previous small bowel follow through demonstrating small bowel CD (37.5% retention risk), and BMI <5th percentile with known IBD (43% retention risk). Caution is advised in these pediatric patients before capsule ingestion. PMID- 19561548 TI - Prebiotics improve gastric motility and gastric electrical activity in preterm newborns. AB - The aim of this double-blind, randomized, placebo-controlled study was to evaluate the effect of a prebiotic mixture on gastric motility in preterm newborns. After a feeding period of 15 days, gastric electrical activity was measured by electrogastrography, and the gastric emptying time was studied by ultrasound technique. No difference was seen in the daily increase of body weight, and no adverse events have been reported. The percentage of time in which propagation was detected in the electrogastrography signal was twice in newborns receiving formula with prebiotics with respect to placebo, and the gastric half emptying time was 30% faster in the prebiotic group than the placebo group. Prebiotic oligosaccharides can modulate the electrical activity and the gastric emptying and may improve the intestinal tolerance of enteral feeding in preterm infants. PMID- 19561549 TI - Autoimmune hepatitis in a child with chronic hepatitis B virus infection. PMID- 19561550 TI - Herpes simplex and eosinophilic oesophagitis: the chicken or the egg? PMID- 19561551 TI - Impact of the ASCO 2007 presentation of HOG Lun 01-24/USO-023 on the prescribing plans of American medical oncologists for patients with stage IIIB non-small cell lung cancer. AB - INTRODUCTION: Nonoperative treatment of stage III non-small cell lung cancer has evolved over the past 30 years. The current approach in the Unites States most often includes concurrent chemoradiotherapy. METHODS: We have used live, case based research events to document prescribing plans among American medical oncologists for first-line therapy in patients with N3 stage IIIB non-small cell lung cancer. Changes in prescribing plans documented before and after the 2007 American Society of Clinical Oncology (ASCO) presentation of a Hoosier Oncology Group trial testing the role of consolidation docetaxel chemotherapy in this setting are presented. RESULTS: Data from 2007 show a post-ASCO shift away from plans for docetaxel consolidation, increased use of concurrent chemoradiotherapy alone, and stable to increased plans for concurrent chemoradiation followed by additional cycles of the chemotherapy used during concurrent management (20%). Preliminary data from 2008 confirm the durability of these changes. CONCLUSIONS: The findings of the Hoosier Oncology Group trial support a transition away from docetaxel consolidation. A trend in this direction among American medical oncologists is clear from our data. However, nearly 20% of oncologists studied in 2008 still plan to use docetaxel consolidation. Furthermore, a majority of those studied after ASCO 2007 continue to report plans to use more than two cycles of chemotherapy as part of their preferred treatment recommendation despite no level I evidence to support this approach. PMID- 19561553 TI - Systematic review: prevalence of malignant incidental thyroid nodules identified on fluorine-18 fluorodeoxyglucose positron emission tomography. AB - PURPOSE: To review current literature and determine the prevalence of malignant incidental focal hypermetabolic thyroid lesions detected by fluorine-18 fluorodeoxyglucose positron emission tomography (F-FDG-PET). MATERIALS AND METHODS: Studies evaluating thyroid carcinomas discovered incidentally on F-FDG PET were systematically searched in the MEDLINE, CINAHL, and Evidence-Based Medicine (EBM) Reviews from 1998 to 2007. Studies were eligible for inclusion with the following criteria: F-FDG-PET studies carried out on humans, long-term clinical follow-up or presence of histological confirmation of thyroid disease, and if studies reported the prevalence of abnormal F-FDG-PET scans with thyroid lesions and confirmed pathological thyroid disease. Studies were excluded if (1) there was no confirmed diagnosis, (2) there was a history of thyroid cancer, (3) they were carried out for diagnoses of thyroid abnormalities, or (4) they were case series. Two reviewers independently reviewed each study's eligibility and abstracted the data. RESULTS: Eighteen articles met criteria resulting in a total of 55 160 patients with 571 patients (1%) having an unexpected focal abnormality in the thyroid gland. Diagnostic confirmations were obtained in 322 patients. Among the confirmed diagnoses, 200 (62.1%) were benign, 107 (33.2%) were malignant, and 15 (4.7%) were indeterminate or a patient without a clear diagnosis. Papillary thyroid carcinoma was the most prevalent thyroid malignancy (82.2%). Eight studies reporting individual maximum standard uptake values were included in a subanalysis. The mean maximum standard uptake value for 73 benign lesions was 4.6+/-2.1, and for the 52 malignant lesions was 6.8+/-4.6 (P<0.001). CONCLUSION: The high prevalence of malignancy associated with focal hypermetabolic thyroid nodules found on F-FDG-PET warrants further evaluation when detected. PMID- 19561552 TI - Predicting the myelotoxicity of chemotherapy: the use of pretreatment O6 methylguanine-DNA methyltransferase determination in peripheral blood mononuclear cells. AB - To assess the value of pretreatment O6-methylguanine-DNA methyltransferase (MGMT) expression in peripheral blood mononuclear cells (PBMCs) in predicting haematological toxicity with O6-alkylating agent chemotherapy, we explored this relationship retrospectively in melanoma patients. Ninety-three patients treated with temozolomide or dacarbazine in four clinical trials were assessed, and a model of the interaction between MGMT expression and haematological toxicity was constructed. Nadir white-cell and platelet counts were related to, and hence could be predicted from, pretreatment MGMT. Leucopenia and thrombocytopenia were more prevalent amongst patients with low pretreatment MGMT, according to the highest grades of toxicity experienced and/or the dose intensity patients could sustain. Addition of interferon to chemotherapy or compression of the temozolomide schedule increased the toxicity. The model also predicts significant myelotoxicity where PBMC MGMT is inactivated, consistent with the experience in the clinic with lomeguatrib and O6-benzylguanine. Determination of MGMT in PBMC can identify patients at greatest risk of toxicity or who are suitable for dose intensification. PMID- 19561554 TI - Presurgical (99m)Tc-sestamibi brain SPET/CT versus SPET: a comparison with MRI and histological data in 33 patients with brain tumours. AB - PURPOSE: A morphofunctional approach to the management of brain tumours has been claimed to increase diagnostic accuracy. Among the proposed single-photon emission tomography (SPET) tracers, (99m)Tc-sestamibi is able to distinguish recurrent tumour from radio-necrosis and to identify early response or resistance to chemotherapy. Major drawbacks of sestamibi, that is, poor morphological resolution and the sites of physiological uptake, could be overcome by dual modality, integrated systems. The purpose of this study was to investigate the real usefulness of (99m)Tc-sestamibi SPET/computed tomography (CT) and to establish a semiquantitative index. METHODS: Charts from 33 consecutive patients selected for surgery, who underwent preoperative SPET/CT and magnetic resonance imaging (MRI), were reviewed. Tumours were confirmed histologically after the surgery in all patients and classified according to WHO recommendations. Semiquantitative indexes were obtained on images (maximum likelihood expectation maximization reconstructed) with and without attenuation correction and visual analysis of SPET versus SPET/CT was performed. RESULTS: A significant statistical difference was shown between SPET and SPET/CT in terms of the delineation of medial shift, oedema and the ability to distinguish tumour from the skull meninges complex and plexus. With regard to semiquantitative indexes, a ratio obtained comparing counts/pixel derived from a region of interest in the tumour area with mirrored region of interest in the contralateral site revealed a sensitivity of 90.9% and specificity of 71.45% in discriminating WHO grade 4 gliomas from a lower grade. CONCLUSION: SPET/CT can distinguish tumour from the skull and other sites of physiological uptake better than SPET alone (as confirmed by MRI in all cases) and affords a morphological map. The proposed semiquantitative index also seems promising in identifying higher-grade disease. SPET/CT thus seems a useful additional tool in brain tumour management, especially when MRI is not feasible or PET/CT is not available. PMID- 19561556 TI - Defining acute lung disease in children with the oxygenation saturation index. AB - OBJECTIVE: To evaluate whether a formula could be derived using oxygen saturation (Spo2) to replace Pao2 that would allow identification of children with acute lung injury and acute respiratory distress syndrome. Definitions of acute lung injury and acute respiratory distress syndrome require arterial blood gases to determine the Pao2/Fio2 ratio of 300 (acute lung injury) and 200 (acute respiratory distress syndrome). DESIGN: Post hoc data analysis of measurements abstracted from two prospective databases of randomized controlled trials. SETTING: Academic pediatric intensive care units. PATIENTS: A total of 255 children enrolled in two large prospective trials of therapeutic intervention for acute lung disease: calfactant and prone positioning. INTERVENTIONS: Data were abstracted including Pao2, Paco2, pH, Fio2, and mean airway pressure. Repeated measures analyses, using linear mixed-effects models, were used to build separate prediction equations for the Spo2/Fio2 ratio, oxygenation index [(Fio2 x Mean Airway Pressure)/Pao2], and oxygen saturation index [(Fio2 x Mean Airway Pressure)/Spo2 ]. A generalization of R was used to measure goodness-of-fit. Generalized estimating equations with a logit link were used to calculate the sensitivity and specificity for the cutoffs of Pao2/Fio2 ratio of 200 and 300 and equivalent values of Spo2/Fio2 ratio, oxygenation index, and oxygen saturation index. MEASUREMENTS AND MAIN RESULTS: An Spo2/Fio2 ratio of 253 and 212 would equal criteria for acute lung injury and acute respiratory distress syndrome, respectively. An oxygenation index of 5.3 would equal acute lung injury criteria, and an oxygenation index of 8.1 would qualify for acute respiratory distress syndrome. An oxygen saturation index, which includes the mean airway pressure and the noninvasive measure of oxygenation, of 6.5 would be equivalent to the acute lung injury criteria, and an oxygen saturation index of 7.8 would equal acute respiratory distress syndrome criteria. CONCLUSIONS: Noninvasive methods of assessing oxygenation may be utilized with reasonable sensitivity and specificity to define acute lung injury and acute respiratory distress syndrome, and, with prospective validation, have the potential to increase the number of children enrolled into clinical trials. PMID- 19561555 TI - A new self-report measure of self-management of type 1 diabetes for adolescents. AB - BACKGROUND: The development of instruments to measure self-management in youth with type 1 diabetes has not kept up with current understanding of the concept. OBJECTIVE: This study aimed to report the development and the testing of a new self-report measure to assess the Self-Management of Type 1 Diabetes in Adolescents (SMOD-A). METHODS: Following a qualitative study, items were identified and reviewed by experts for content validity. A total of 515 adolescents, 13 to 21 years old, participated in a field study by completing the SMOD-A (either once or twice) and additional measures of diabetes-related self efficacy (Self-Efficacy for Diabetes Scale), quality of life (Diabetes Quality of Life for Youth Questionnaire), self-management (Diabetes Self-Management Profile), and adherence (Self-Care Inventory). Data were collected also on metabolic control (glycosylated hemoglobin [HbA1c]). RESULTS: The content validity index was .93. Exploratory alpha factor analyses revealed five subscales: Collaboration With Parents, Diabetes Care Activities, Diabetes Problem Solving, Diabetes Communication, and Goals (alpha = .71 to .85). The stability of the SMOD-A ranged from .60 to .88 at 2 weeks (test-retest) to .59 to .85 at 3 months. Correlations of SMOD-A subscales with Self-Efficacy for Diabetes Scale Diabetes; Diabetes Quality of Life for Youth Questionnaire satisfaction, impact, and worry; Diabetes Self-Management Profile; and Self-Care Inventory were generally significant and in the expected direction. Collaboration with parents and HbA1c values were related significantly and positively (r = .11); all other SMOD-A subscales were related significantly and negatively to HbA1c (r = -.10 to .26), demonstrating that better self-management is associated somewhat with better metabolic control and supporting construct validity of the new measure. DISCUSSION: The SMOD-A has been found to be a reliable, stable, and valid measure of SMOD-A. PMID- 19561557 TI - Activated recombinant factor VII for refractory bleeding during extracorporeal membrane oxygenation. AB - OBJECTIVE: To determine the frequency of adverse events with the use of activated recombinant factor VII during extracorporeal membrane oxygenation support and to quantify the effect on bleeding parameters. DESIGN: Retrospective case series from January 1999 to August 2006. SETTING: Pediatric intensive care unit at a tertiary academic children's hospital. PATIENTS: Seventeen patients received a total of 26 doses of activated recombinant factor VII while supported on extracorporeal membrane oxygenation or within 3 hrs of initiation of extracorporeal membrane oxygenation support from February 2003 to August 2006, and 23 historical controls from January 1999 to December 2002 with bleeding complications reported to the Extracorporeal Life Support Organization database while supported on extracorporeal membrane oxygenation before the use of activated recombinant factor VII. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: No significant difference in the rate of thromboembolic complications, extracorporeal membrane oxygenation circuit failures, or mortality was found between the patients and historical controls. No trend toward increased survival was found, and a significant number of circuit complications was seen in both groups. In patients treated with activated recombinant factor VII, a significant reduction in chest tube output and blood product transfusion rates was seen within 5 hrs of activated recombinant factor VII administration. CONCLUSION: Activated recombinant factor VII administration during extracorporeal membrane oxygenation support was associated with a decrease in bleeding severity (indicated by chest tube output and blood product transfusion rates) and was not associated with an increased rate of thromboembolic complications. PMID- 19561558 TI - Failure to recover somatotropic axis function is associated with mortality from pediatric sepsis-induced multiple organ dysfunction syndrome. AB - OBJECTIVES: To describe the associations between mediators of the somatotropic axis and mortality from sepsis-induced multiple organ dysfunction syndrome in children; and to examine the relationship between immune function and the somatotropic axis in this setting. DESIGN: Retrospective study using banked plasma. SETTING: Single mixed surgical/medical intensive care unit at a quaternary level children's hospital. PATIENTS: A total of 24 children (n = 17 survivors, 7 nonsurvivors) with severe sepsis or septic shock and dysfunction of >or=2 organ systems. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Plasma samples were available from days 3, 7, and 14 of multiple organ dysfunction syndrome. Insulin-like growth factor 1 and insulin-like growth factor binding protein 3 levels were measured by chemiluminescence. Immune function was quantified using previously determined ex vivo lipopolysaccharide-induced tumor necrosis factor-alpha production levels and absolute lymphocyte counts. Insulin like growth factor 1 levels were lower in nonsurvivors compared with survivors (p = .004) with the greatest difference seen on day 14 (25 [25-69] ng/mL vs. 314 [123-582] ng/mL; p = .038). insulin-like growth factor binding protein 3 levels were reduced similarly over time (p = .019). A drop in plasma insulin-like growth factor binding protein 3 level at any time after day 3 of illness resulted in a 35-fold increased odds of death (odds ratio, 35 [1.6-750]; p = .004). Both ex vivo tumor necrosis factor-alpha and absolute lymphocyte count were reduced in nonsurvivors compared with survivors, but these differences occurred earlier (days 3 and 7). CONCLUSIONS: These data suggest that prolonged reduction of somatotropic axis function is associated with mortality from pediatric sepsis induced multiple organ dysfunction syndrome. Reductions in innate and adaptive immune function are common in this population and are associated with failure of recovery of the somatotropic axis, although the nature of these relationships remains incompletely understood. PMID- 19561560 TI - Massage for low back pain: an updated systematic review within the framework of the Cochrane Back Review Group. AB - STUDY DESIGN: Systematic Review. OBJECTIVES: To assess the effects of massage therapy for nonspecific low back pain. SUMMARY OF BACKGROUND DATA: Low back pain is one of the most common and costly musculoskeletal problems in modern society. Proponents of massage therapy claim it can minimize pain and disability, and speed return to normal function. METHODS: We searched MEDLINE, EMBASE, CINAHL from their beginning to May 2008. We also searched the Cochrane Central Register of Controlled Trials (The Cochrane Library 2006, issue 3), HealthSTAR and Dissertation abstracts up to 2006. There were no language restrictions. References in the included studies and in reviews of the literature were screened. The studies had to be randomized or quasi-randomized trials investigating the use of any type of massage (using the hands or a mechanical device) as a treatment for nonspecific low back pain. Two review authors selected the studies, assessed the risk of bias using the criteria recommended by the Cochrane Back Review Group, and extracted the data using standardized forms. Both qualitative and meta-analyses were performed. RESULTS: Thirteen randomized trials were included. Eight had a high risk and 5 had a low risk of bias. One study was published in German and the rest in English. Massage was compared to an inert therapy (sham treatment) in 2 studies that showed that massage was superior for pain and function on both short- and long-term follow-ups. In 8 studies, massage was compared to other active treatments. They showed that massage was similar to exercises, and massage was superior to joint mobilization, relaxation therapy, physical therapy, acupuncture, and self-care education. One study showed that reflexology on the feet had no effect on pain and functioning. The beneficial effects of massage in patients with chronic low back pain lasted at least 1 year after the end of the treatment. Two studies compared 2 different techniques of massage. One concluded that acupuncture massage produces better results than classic (Swedish) massage and another concluded that Thai massage produces similar results to classic (Swedish) massage. CONCLUSION: Massage might be beneficial for patients with subacute and chronic nonspecific low back pain, especially when combined with exercises and education. The evidence suggests that acupuncture massage is more effective than classic massage, but this need confirmation. More studies are needed to confirm these conclusions, to assess the impact of massage on return-to-work, and to determine cost-effectiveness of massage as an intervention for low back pain. PMID- 19561561 TI - Bone graft volumetric changes and clinical outcomes after instrumented lumbar or lumbosacral fusion: a prospective cohort study with a five-year follow-up. AB - STUDY DESIGN: Prospective cohort study. OBJECTIVE: To determine: (1) patterns of bone graft volumetric changes, (2) whether different fusion methods, numbers of fusion segments, and range of fusion affect bone graft volumetric changes, and (3) whether these changes affect clinical and functional outcomes after surgery. SUMMARY OF BACKGROUND DATA: Bone graft volumetric change is likely to importantly determine successful fusion. However, despite the importance of graft volumetric changes, little information is available on bone graft volumetric changes after instrumented fusion. METHODS: Thirty-one patients with lumbosacral disease who underwent decompression and instrumented spinal fusion using autologous iliac bone graft were included in this study. Patients were classified into 3 major categories, based on fusion methods, the number of fusion segments, and range of fusion, namely, (1) the posterolateral fusion and the posterior lumbar interbody fusion groups, (2) the one-segment fusion and 2-segment fusion groups, and (3) the floating fusion (fusion to L5) and fixed fusion (fusion to S1) groups. To evaluate bone graft volumetric changes, computed tomography scans were performed. Group graft volumes were compared, and Oswestry Disability Index and visual analogue scales scores were obtained to evaluate clinical and functional outcomes. RESULTS: Initial graft volumes had decreased by 8% at 6 months and by 26.3% between 6 and 12 months after surgery (P = 0.0025, P = 0.0001, respectively). Mean group bone graft volume losses at 5 years after surgery were not significantly different between the posterolateral fusion and posterior lumbar interbody fusion groups or the floating fusion and fixed fusion groups (P = 0.722, P = 0.076, respectively). However, mean bone graft volume loss was significant greater in the 2-segment fusion group than in the one-segment fusion group (P = 0.007). No statistical difference in visual analogue scales and Oswestry Disability Index score decreases were observed between the groups. Mean graft bone volume loss during 5 years increased with initial graft volume (r = 0.525, P = 0.003). However, ratios of volume loss over 5 years versus initial graft volume decreased with initial graft volume (r = -0.459, P = 0.011). CONCLUSION: One-third of initial bone grafts were found to be resorbed over 5 years and the majority of this resorption was found to occur between 6 months and 1 year after instrumented lumbar/lumbosacral fusion. Furthermore, bone graft volumetric changes were not affected by fusion methods or ranges of fusion, and did not affect clinical or functional outcomes. Although bone graft volume loss was found to increase with initial graft volume, larger bone grafts were found to result in a larger fusion mass. PMID- 19561566 TI - Bisphosphonates and dental implants: current problems. AB - Osteonecrosis of the jaw has been described in patients taking bisphosphonates after oral surgery procedures, including the placement of dental implants. This review is an update of the relationship between bisphosphonates and dental implants. Results obtained by different authors are compared, contrasting earlier studies where an improvement in implant osseointegration using bisphosphonates was observed, with ones where statistically significant differences were found, and more recent studies disagreeing with the use of bisphosphonates for causing necrosis of the jaw. The differing results obtained between animal studies and the situation observed in humans may be due to a short medication and follow-up period, as well as to the existence of few research studies where dental implants are placed in the oral cavity. Currently, dental implants are contraindicated in patients being treated with intravenous bisphosphonates. In 2007, the American Association of Oral and Maxillofacial Surgeons suggested guidelines for patients treated with oral bisphosphonates, based on the clinical situation of the patient and the length of treatment with the drug, and that greater caution prior and subsequent to surgery should be taken for three years after treatment. All patients treated with bisphosphonates must have the risk of possible loss of implants and the risk of suffering a bony necrosis of the operated jaw explained to them, and give their informed consent prior to dental implant surgery. PMID- 19561562 TI - Patient-level minimal clinically important difference based on clinical judgment and minimally detectable measurement difference: a rationale for the SF-36 physical function scale in the SPORT intervertebral disc herniation cohort. AB - STUDY DESIGN: A proof of concept case study. OBJECTIVE: To introduce and evaluate a method for identifying what constitutes a minimal clinically important difference (MCID) in the SF-36 Physical Function scale at the patient level. SUMMARY OF BACKGROUND DATA: MCID has become increasingly important to researchers interested in evaluating patient care. Over the last 30 years, an array of approaches for assessing MCID has evolved with little consensus on which approach applies in any given situation. METHODS: Three approaches for estimating standard errors of measurement (se) and a 30% change approach for establishing MCID were evaluated for the physical function (PF) scale with SPORT patients in the intervertebral disc herniations cohort. MCIDs for each se approach were then developed based on (1) these standard errors and (2) clinically relevant factors including: (a) baseline PF score and (b) acceptable risk for type I error. RESULTS.: Intervertebral disc herniations patients (N = 996) identified from the SPORT database met inclusion criteria. The se for the classic test theory (CTT) based test level approach was 9.66. CTT-score-level and IRT-pattern-level standard errors varied depending on the score, and ranged between (2.73-7.17) and (5.96-16.2), respectively. As predicted, CTT-score-level se values were much smaller than IRT-pattern-level se values at the extreme scores and IRT-pattern level se values were slightly smaller than CTT score-level se values in the middle of the distribution. Across follow-up intervals, the CTT-score-based approach consistently demonstrated greater sensitivity for identifying patients who were improved or worsened. Comparisons of CTT-based-score-level se and 30% improvement rule MCID estimates were as hypothesized: MCID values for 30% gains demonstrated substantially lower sensitivity to change for baseline PF scores in the 0 to 50 range but were similar to CTT-score-level-based MCIDs when baseline scores were above 50. CONCLUSION: The CTT-based-score-level approach for establishing MCID based on the clinical relevance of the baseline PF score and the tolerance for erroneously accepting an observed change as reliable provided the more sensitive and theoretical compelling approach for estimating MCID at the patient level, which in turn will provide fundamentally important to the clinician regarding treatment efficacy at the patient level. PMID- 19561567 TI - An interview with Nobel laureate Roy Glauber, Physics 2005. AB - The field of quantum optics rests on the work of Roy Glauber, who helped elucidate the nature of light as both particles and waves. According to Glauber, quantum optics allowed "all sorts of experiments...that never could have been done before." He suggests that it was not his "small revelation" that the Nobel Committee awarded, but rather the decades of research that followed his own. Nonetheless, Glauber received one-half of the 2005 Nobel Prize in Physics "for his contribution to the quantum theory of optical coherence" while the other half was shared by John Hall and Theodor Hansch for their work on laser-based precision spectroscopy. Glauber admits that the behavior of light seems strange and unintuitive--yet the phenomena that Einstein called "spooky action at a distance" may have many practical applications. In this candid interview, Glauber shares his thoughts about working at Los Alamos National Laboratory--his shock to learn that he was helping to build The Bomb, and his dismay about how it was used. At Los Alamos, Glauber met two of his major influences: Julian Schwinger, who was Glauber's thesis advisor at Harvard, and Los Alamos scientific director Robert Oppenheimer, who facilitated his early post-doctoral research. Glauber also tells a poignant account of how his marriage fell victim to the social upheaval of the 1960's, and how he was left to raise two children alone. Despite the difficulties of reconciling academia with family, Glauber is amused to find himself revered by women as "someone who has raised children and nonetheless had a successful academic career." PMID- 19561568 TI - Detection and characterization of EWSR1/ATF1 and EWSR1/CREB1 chimeric transcripts in clear cell sarcoma (melanoma of soft parts). AB - Unlike melanoma, clear cell sarcoma harbors either a t(12;22)(q13;q12) recurrent translocation, resulting in an EWSR1/ATF1 chimeric gene, or less commonly a t(2;22)(q34;q12) translocation fusing EWSR1 and CREB1. Few studies have examined the prevalence of all chimeric types and variants to assess the usage of ancillary genetic testing in routine diagnosis. We investigated rearrangement prevalence in 17 clear cell sarcomas, two positive control cell lines, and two melanomas (negative controls). Fluorescence in situ hybridization (FISH) analysis using the LSI EWSR1 break-apart probe and a reverse transcription polymerase chain reaction (RT-PCR) assay optimized for formalin-fixed paraffin-embedded tissue to detect all four reported EWSR1/ATF1 clear cell sarcoma chimeric types and the EWSR1/CREB1 variant was performed. All 15 cases available for testing by FISH were positive for EWSR1 rearrangement including two cases with insufficient RNA for RT-PCR. Thirteen of 15 cases successfully tested by RT-PCR harbored a type 1 chimeric transcript (EWSR1 exon 8/ATF1 exon 4), of which five tumors simultaneously carried a type 2 chimeric transcript (EWSR1 exon 7/ATF1 exon 5). One case carried a type 2 transcript alone and one case contained an EWSR1/CREB1 transcript. Both control cases were positive by both techniques with one case carrying both types 1 and 2 chimeric transcripts and the other types 2 and 3 (EWSR1 exon 10/ATF1 exon 5). Consequently, both techniques are equally effective in assessing for an EWSR1 rearrangement and are useful ancillary diagnostic tests for clear cell sarcoma. They also reinforce the prevalence of this translocation in these tumors. In addition, EWSR1-CREB1 was identified in a clear cell sarcoma of soft tissue providing further evidence that this chimeric variant is not exclusive to gastrointestinal clear cell sarcomas and should be included in RT PCR assays of soft tissue clear cell sarcomas. PMID- 19561569 TI - New readers start here. PMID- 19561570 TI - Polyol-containing chewing gums: flawed evidence. PMID- 19561571 TI - Early childhood caries--risk factors. AB - DESIGN: This was an Australian case-control study. CASE-CONTROL SELECTION: Cases were recruited from children referred for dental treatment under general anaesthesia at free public hospitals in eight health service districts in the state of Queensland, Australia [early childhood caries (ECC) public cases], and three private specialist paediatric dental clinics (ECC private cases). Controls were selected from a full list of all childcare facilities in the area using a selection ratio of one in seven children. As dental health status of the children was unknown prior to recruitment, a subgroup of 62 children with ECC was recruited in the control cohort (ECC childcare) and formed the third source of ECC cases. ASCERTAINMENT: The teeth of children in dental clinics or childcare facilities were examined using lighting from an examiner's head-lamp, with the child placed on the laps of the mother and examiner. A child was considered to have ECC if at least one cavity was present. Caries was charted using the World Health Organization oral health survey basic methods criteria(1) and enamel hypoplasia using the modified Developmental Defects of Enamel index. Presence of Streptococcus mutans was also assessed. Mothers were interviewed and screened to determine their social, medical and dental histories; dental caries experience; absence or presence of plaque and gingival inflammation; and presence of S. mutans. Validated questionnaires were used to obtain social, medical, dental, dietary and toothbrushing histories of the mothers. DATA ANALYSIS: Group comparisons of continuous variables (such as age and birthweight) were compared for statistical significance using analysis of variance. Categorical variables were compared for statistical difference across groups using contingency chi2 tests together with multinomial logistic regression modelling. RESULTS: A large proportion of children tested positive for S. mutans if their mothers also tested positive. A common risk indicator found in ECC children from childcare facilities and public hospitals was visible plaque [odds ratio (OR), 4.1; 95% confidence interval (CI), 1.0-15.9; and OR, 8.7; 95% CI, 2.3-32.9, respectively). Compared with ECC-free controls, the risk indicators specific to childcare cases were enamel hypoplasia (OR, 4.2; 95% CI, 1.0-18.3), difficulty in cleaning the child's teeth (OR 6.6; 95% CI, 2.2-19.8), presence of S. mutans (OR, 4.8; 95% CI, 0.7 32.6), sweetened drinks (OR, 4.0; 95% CI, 1.2-13.6) and maternal anxiety (OR, 5.1; 95% CI, 1.1-25.0). Risk indicators specific to public hospital cases were presence of S. mutans in the child (OR, 7.7; 95% CI, 1.3-44.6) or mother (OR, 8.1; 95% CI, 0.9-72.4), ethnicity (OR, 5.6; 95% CI, 1.4-22.1), and access of mother to pension or healthcare card (OR, 20.5; 95% CI, 3.5-119.9). By contrast, a history of chronic ear infections was found to be protective for ECC in childcare children (OR, 0.28; 95% CI, 0.09-0.82). CONCLUSIONS: This case-control study showed that children of different socioeconomic backgrounds who have ECC share the common risk indicators of visible plaque, consumption of sugary snacks and presence of S. mutans. Additional risk indicators in children from childcare facilities were enamel hypoplasia, difficulty in cleaning the child's teeth, sweetened drinks and maternal anxiety, whereas ethnicity and mothers' access to pension or healthcare cards were specific to the public hospital cases. PMID- 19561572 TI - Manual and electronic probes have similar reliability in the measurement of untreated periodontitis. AB - DATA SOURCES: The Cochrane Central Register of Controlled Trials, PubMed Medline and Latin American and Caribbean Health Science literature databases were searched, and searches made by hand of identified papers, to source relevant data. (BIOSIS Previews) were searched. There were no language restrictions. STUDY SELECTION: Studies were evaluated by two reviewers independently. Only clinical trials were included that were published in the English, Spanish and Portuguese languages and were performed on humans. Articles were excluded when the examiners were not calibrated; when they did not compare manual and electronic probing techniques; when they did not measure the clinical attachment level (CAL); and when the subjects did not present destructive periodontal disease or had already received periodontal treatment. DATA EXTRACTION AND SYNTHESIS: The quality of the identified studies was assessed and standardised data extracted. Only two studies met all the selection criteria so no meta-analysis was performed. RESULTS: Only two of the 37 identified articles were included in the review. The results of these two studies showed that the mean variance and the absolute mean difference between CAL measurements for the two types of probes were not statistically different. CONCLUSIONS: Manual and electronic probes were of similar reliability when used to measure CAL in individuals who had untreated periodontitis and when used by a calibrated examiner, but this finding is not supported by strong evidence. PMID- 19561573 TI - Is oral irrigation beneficial to gingival health as an adjunct to toothbrushing? AB - DATA SOURCES: Relevant papers were identified by using Medline-PubMed and the Cochrane Central Register of Controlled Trials. STUDY SELECTION: Papers were assessed for inclusion independently by two reviewers and only those published in the English language were chosen. Randomised controlled clinical trials (RCT) or controlled clinical trials (CCT) conducted in adults with good general health were selected. OUTCOME MEASURES: Evaluation parameters included changes in dental plaque, gingival bleeding/ inflammation and probing pocket depth. Study quality was assessed based upon study design, evaluation period, profiles of subjects, method of randomisation, blindness of examiners and completeness of followup assessment. DATA EXTRACTION AND SYNTHESIS: Data were extracted by three reviewers. As the selected studies showed considerable heterogeneity in the study designs, characteristics, outcome variables and results, a descriptive review of the data was presented instead of performing a meta-analysis. RESULTS: Seven papers (four RCT and three CCT) were included. Mean values and standard deviations were collected from the data. Data showed that oral irrigation does not reduce plaque scores more than toothbrushing or regular oral hygiene measures. CONCLUSIONS: This systematic review suggests that oral irrigation as an adjunct to toothbrushing does not have a beneficial effect in reducing dental plaque, but it may improve gingival health. PMID- 19561574 TI - Positive relationship between stress and periodontal disease? AB - DATA SOURCES: Medline and the Cochrane Oral Health Group Specialist Trials Register were utilised to find relevant studies. STUDY SELECTION: Articles were examined independently by two reviewers, and all review articles and animal studies were excluded. Studies published in the English language were included if they controlled for the potential effect of confounding factors, had adequate criteria to define periodontal disease, and it was possible to establish evaluate stress levels and the methodological quality of the study. DATA EXTRACTION AND SYNTHESIS: Data were extracted by two reviewers independently and verified by a third. A qualitative summary of the findings was presented. RESULTS: One prospective clinical trial, seven case-control studies and six cross-sectional studies were included. Eight studies found a positive outcome between psychosocial factors or stress and periodontal disease; four studies observed a positive outcome for some characteristics and a negative outcome for others; whereas two studies found a negative outcome between psychosocial factors or stress and periodontal disease. CONCLUSIONS: Within the limitations of this systematic review, the majority of studies showed a positive relationship between stress or psychological factors and periodontal disease. In the future, however, well-designed and more representative studies should be considered to confirm these factors as a risk for periodontal disease. PMID- 19561575 TI - Do woodsticks work? AB - DATA SOURCES: Medline and the Cochrane Central Register of Controlled Trials were used to find relevant studies. STUDY SELECTION: Studies were screened independently by two reviewers. Randomised controlled trials and controlled clinical trials were selected if they were conducted in individuals of over 18 years of age who were in good general health, and which used plaque, bleeding or gingivitis as outcome measures. Case reports, letters, and narrative or historical reviews were excluded and only English-language papers were considered. DATA EXTRACTION AND SYNTHESIS: Because of the heterogeneity of the studies' designs, a qualitative summary was presented. RESULTS: Seven publications describing eight clinical experiments met the inclusion criteria. The improvement in gingival health observed in the studies represented a significant reduction of bleeding realised by the use of triangular woodsticks. None of the studies that scored plaque demonstrated any significant advantage of the use of woodsticks over alternative methods of plaque removal in people who had gingivitis. CONCLUSIONS: The evidence from clinical controlled trials shows that woodsticks do not have an additional effect on visible interdental plaque, but do, however, help improve interdental gingival inflammation. PMID- 19561576 TI - Cetylpyridinium chloride-containing mouth rinses and plaque control. AB - DATA SOURCES: Medline and the Cochrane Central register of controlled trials were used to identify relevant reports. STUDY SELECTION: Randomised controlled trials and controlled clinical trials were included if they were of >/= 4 weeks' duration, were conducted in people of >/= 18 years of age and in good general health, and used plaque/ bleeding/ gingivitis or pocket depth as outcome measures. Case reports, letters and narrative or historical reviews were excluded and only English language papers were considered. DATA EXTRACTION AND SYNTHESIS: Descriptive comparisons are presented for brushing only or for brushing and rinsing and, where appropriate, a meta-analysis was performed using a random effects model. RESULTS: Eight studies met the inclusion criteria and they showed a small but significant additional benefit of cetylpyridinium chloride (CPC) use. CONCLUSIONS: The existing evidence supports the use of CPC-containing mouthrinses as adjuncts to either supervised or unsupervised oral hygiene as they provide a small but significant additional benefit in reducing plaque accumulation and gingival inflammation. PMID- 19561577 TI - Which type of scaling system is best? AB - DATA SOURCES: Medline and the Cochrane Central register of Controlled Trials were searched for relevant studies. STUDY SELECTION: Randomised controlled trials and controlled clinical trials conducted in vitro or in vivo on human teeth, which used plaque, bleeding, gingivitis or pocket depth, clinical attachment level, microbiological findings, patient perception and treatment time as outcome measures, were included. Case reports, letters and narrative or historical reviews were excluded and only English-language papers were considered. DATA EXTRACTION AND SYNTHESIS: The studies included differed in design and outcome so a qualitative summary was presented. RESULTS: Fifteen studies met the inclusion criteria. The Vector (Durr Dental, Bietigheim-Bissingen, Germany) ultrasonic scaler (VUS) provided clinical and microbiological periodontal healing results comparable to scaling and root planing and a conventional ultrasonic system in moderately deep pockets. CONCLUSIONS: The VUS may be used as a gentle root debridement device for supportive periodontal therapy, as an alternative to other conventional ultrasonic systems. The operator should consider, however, the extra time needed for instrumentation. PMID- 19561578 TI - Oral hygiene in elderly people in hospitals and nursing homes. AB - DATA SOURCES: Searches were made using Medline, the Cochrane Library, the National Health Service Economic Evaluation database and, by hand, of relevant articles' reference lists. STUDY SELECTION: The search was limited to studies conducted in humans only. The predetermined inclusion criteria were: clinical studies and randomised controlled trials (RCT) that linked oral hygiene with healthcare-associated pneumonia or respiratory tract infection in elderly people. Publications in Dutch, English, German and any of the Nordic languages (Danish, Finnish, Icelandic, Norwegian, Swedish) were included. Articles about authority opinions and reports of expert committees were excluded, as were studies on subjects who required mechanical ventilation or tube feeding. DATA EXTRACTION AND SYNTHESIS: Data extraction from RCT was focused on the outcomes. Assessments were made of the quality and validity of the studies, using statistical methods to test these. To ensure the consistency of the assessments throughout the study, two authors (EN, PS) performed the data extraction independently, and any disagreements were resolved in consensus meetings. Pooling data from individual studies (meta-analysis) was not deemed appropriate because of heterogeneous study designs, quality of reporting methodological aspects and trial conduct. To provide an overview of additional clinical studies in this research area, the non RCT studies that were identified were scrutinised for the authors' main conclusion(s). RESULTS: Fifteen publications, of which five were RCT, fulfilled the inclusion criteria and were considered throughout all the assessments. All of the RCT revealed positive preventive effects of oral care on pneumonia or respiratory tract infection in nursing home residents, with numbers needed to treat ranging from 8.6-15.3. CONCLUSIONS: Available results from RCT provide evidence that mechanical oral hygiene decreases mortality risk from pneumonia and seems to have a clinically relevant preventive effect on nonfatal pneumonia in independent elderly individuals. The data show that providing mechanical oral hygiene may prevent approximately 1 in 10 cases of death from pneumonia in dependent elderly people and indicate a largely similar effect for prevention of pneumonia. PMID- 19561579 TI - Vacuum-formed retainers more effective than Hawley retainers. AB - DESIGN: A randomised controlled trial (RCT) was performed in a single orthodontic practice. INTERVENTION: The aim was to compare the clinical effectiveness of Hawley and vacuum-formed retainers (VFR) over a 6-month period of retention. Three hundred and ninety-seven patients were treated by a specialist orthodontist and randomly allocated to wear either Hawley retainers (n=196) or VFR (n=201). OUTCOME MEASURE: A blinded examiner analysed the records of maxillary and mandibular dental casts at debond and at 6 months into retention, assessing tooth rotation mesial to the first permanent molar, intercanine and intermolar widths, and Little's Index of Irregularity. RESULTS: Three hundred and fifty-five individuals participated through to the conclusion of the trial, giving a completion rate of 89%. The results showed significantly greater changes in irregularity of the incisors in the Hawley group than in the VFR group at 6 months. CONCLUSIONS: VRF are more effective than Hawley retainers at holding the correction of the maxillary and mandibular labial segments. PMID- 19561580 TI - Do lip bumpers work? AB - DATA SOURCES: Searches were made using PubMed, Embase, the Cochrane Central Register of Controlled Trials and the Cochrane database of systematic reviews, along with searches by hand of the following journals: Angle Orthodontist, American Journal of Orthodontics and Dentofacial Orthopedics, Journal of Orthodontics and European Journal of Orthodontics. STUDY SELECTION: The first step was to identify eligible reports based on their titles and abstracts: 52 articles were found. Inclusion and exclusion criteria were then applied. Studies were included if subjects were human; they were randomised controlled trials and prospective or retrospective studies; they discussed the effect of the lip bumper on the arch and teeth; and they were reported in the English language. Articles excluded were mainly animal studies, case reports, case series, review articles, abstracts, in-vitro studies, discussions and interviews; were published in a language other than English; or did not follow the objective of this review. The selection was made by two researchers. Their results were compared to identify discrepancies and reach mutual agreement. DATA EXTRACTION AND SYNTHESIS: If two or more studies had evaluated the same technique, a meta-analysis was planned. RESULTS: A total of 16 RCT were identified of which 13 underwent detailed evaluation; of these, only one study was considered to have usable outcome information. The others were excluded because of control group, sex, age, study casts and cephalometric analysis, and measurement error. This single small study showed that lip bumper can increase arch dimensions and contribute to crowding relief in mixed dentition. CONCLUSIONS: The results showed increases in arch dimensions, including a greater arch length. This was attributed to incisor proclination, distalisation, and distal tipping of the molars. There were also increases in arch width and intercanine and deciduous intermolar or premolar distances. The long-term stability of the effects of the lip bumper need to be elucidated. PMID- 19561581 TI - Medication and tooth movement. AB - DATA SOURCES: PubMed, Embase, Web of Science and Biosis were used to search for relevant material, along with the reference lists of retrieved articles. STUDY SELECTION: Both animal and clinical studies were included if they met the following criteria: they had study and control groups of at least five subjects each; used drugs or supplements with known effects on bone physiology; recorded drug dose and administration; gave details of the application of the forces used for tooth movement; described the technique used to measure the rate of tooth movement; and presented a statistical analysis of the results of the study. DATA EXTRACTION AND SYNTHESIS: A formal data extraction process is not described. RESULTS: Forty-nine studies were included in this review. Numerous problems were found in the information presented in the reviewed literature from the almost exclusively animal studies. Comparison of the data from these studies was difficult because of the variability in experimental design, animal models, administration regimens, application and duration of forces to the teeth involved. CONCLUSIONS: The authors identified a need for more well designed studies on the effects of various types of medication on orthodontic tooth movement. PMID- 19561582 TI - Continuing warfarin therapy does not increased risk of bleeding for patients undergoing minor dental procedures. AB - DATA SOURCES: Medline, Embase, Cochrane Central Register of Controlled Trials, International Association for Dental Research abstracts and reference lists of retrieved articles were searched, and contact made with content experts. STUDY SELECTION: Studies were selected independently by two reviewers. Randomised controlled trials (RCT) were selected if: they compared the effects of continuing a regular dose of warfarin with the effects of discontinuing (or modifying) the dose on the incidence of bleeding; the study group participants were people undergoing dental procedures who also had thromboembolism (arterial or venous); and the outcome assessed was postoperative bleeding (major, clinically significant nonmajor, or minor). Study quality was assessed using the Jadad scale. DATA EXTRACTION AND SYNTHESIS: Data extraction was carried out by three reviewers independently. Meta-analysis was conducted using a random-effects model. RESULTS: Five RCT (553 patients) met the inclusion criteria. Compared with interrupting warfarin therapy (either partial or complete), peri-operative continuation of warfarin at the patient's usual dose was not associated with an increased risk for clinically significant nonmajor bleeding [relative risk (RR), 0.71; 95% confidence interval (CI), 0.39-1.28; P 0.65; I(2) 0%) or an increased risk for minor bleeding (RR, 1.19; 95% CI, 0.90-1.58; P 0.22; I(2) 0%). CONCLUSIONS: Continuing the regular dose of warfarin therapy does not seem to confer an increased risk of bleeding compared with discontinuing or modifying the warfarin dose in people undergoing minor dental procedures. PMID- 19561583 TI - The therapeutic use of botulinum toxin in cervical and maxillofacial conditions. AB - DATA SOURCES: Medline, Cochrane Library and bibliographies of identified articles. STUDY SELECTION: Studies of high methodological quality (systematic reviews or randomised controlled trials; RCT) were selected if considered use of botulinum toxin (BTX) either prophylactically or therapeutically, as an adjunct to dental implant therapy for temporomandibular disorders or for other maxillofacial conditions such as bruxism, masseteric hypertrophy, oromandibular dystonia or cervical dystonia. DATA EXTRACTION AND SYNTHESIS: Only studies with high levels of evidence were evaluated. Four RCT met the search criteria in the area of cervical dystonia and chronic facial pain. RESULTS: No RCT were identified that evaluated dental implant therapy along with use of BTX. Four RCT did meet the search criteria in the area of cervical dystonia and chronic facial pain. People who had cervical dystonia exhibited significant improvements in baseline functional, pain and global assessments compared with placebo, whereas individuals with chronic facial pain improved significantly compared with placebo in terms of pain. CONCLUSIONS: During the period of study, no references were found evaluating use of BTX in dental implantology. Nevertheless, it appears relatively safe and effective in treating cervical dystonia and chronic facial pain associated with masticatory hyperactivity. PMID- 19561584 TI - Chlorhexidine gel reduces incidence of alveolar osteitis after extraction of the mandibular third molars. AB - DESIGN: A randomised, prospective clinical trial with parallel groups was carried out in a single centre. INTERVENTION: The experimental (gel) group (n=41) applied a bioadhesive 0.2% chlorhexidine gel to the wound during the first postoperative week and a control (rinse) group (n=32) used a 0.12% (v/v) chlorhexidine mouthrinse during the first week following third molar extraction. Patients were evaluated on the third and seventh postoperative day. OUTCOME MEASURE: Alveolar osteitis was evaluated according to Blum's criteria.. RESULTS: A 70% decrease in postoperative alveolar osteitis in the gel group (P 0.04) was observed. The rinse group had 25% incidence of postoperative alveolar osteitis, whereas the gel group had 7.5%. T equates to a number needed to treat of six (95% confidence interval, 3-144). CONCLUSIONS: It was concluded that the topical application of bioadhesive chlorhexidine gel to the surgical wound during the postoperative week may decrease the incidence of alveolar osteitis after extraction of the mandibular third molars. PMID- 19561585 TI - Inconsistent evidence for the use of tricyclic antidepressants in the treatment of temporomandibular joint disorders. AB - DATA SOURCES: Medline and the Cochrane Library were consulted to find evidence in the literature. STUDY SELECTION: The English language studies identified were considered and stratified according to the Strength of Recommendation Taxonomy (SORT) criteria. DATA EXTRACTION AND SYNTHESIS: A qualitative review was conducted. RESULTS: 11 articles were identified, seven of which were reviews. CONCLUSIONS: There is evidence, based on inconsistent or limited-quality patient oriented data (SORT level B recommendation), for the use of tricyclic antidepressants (TCA) in the treatment of temporomandibular joint disorders (TMD). PMID- 19561586 TI - All right, SQUIRE? PMID- 19561587 TI - American Dental Association evidence-based dentistry website. AB - In March 2009, the American Dental Association (ADA) launched a new evidence based dentistry (EBD) website (ebd.ada.org). The site has been established to provide the dental profession with access to evidence from high-quality systematic reviews and is supported by a grant from the National Library of Medicine and the National Institute for Dental and Craniofacial Research(Grant Number G08 LM008956). PMID- 19561589 TI - Conditional mouse lung cancer models using adenoviral or lentiviral delivery of Cre recombinase. AB - The development of animal models of lung cancer is critical to our understanding and treatment of the human disease. Conditional mouse models provide new opportunities for testing novel chemopreventatives, therapeutics and screening methods that are not possible with cultured cell lines or xenograft models. This protocol describes how to initiate tumors in two conditional genetic models of human non-small cell lung cancer (NSCLC) using the activation of oncogenic K-ras alone or in combination with the loss of function of p53. We discuss methods for sporadic expression of Cre in the lungs through engineered adenovirus or lentivirus, and provide a detailed protocol for the administration of the virus by intranasal inhalation or intratracheal intubation. The protocol requires 1-5 min per mouse with an additional 30-45 min to set-up and allow for the recovery of mice from anesthesia. Mice may be analyzed for tumor formation and progression starting 2-3 weeks after infection. PMID- 19561590 TI - Predicting the effects of coding non-synonymous variants on protein function using the SIFT algorithm. AB - The effect of genetic mutation on phenotype is of significant interest in genetics. The type of genetic mutation that causes a single amino acid substitution (AAS) in a protein sequence is called a non-synonymous single nucleotide polymorphism (nsSNP). An nsSNP could potentially affect the function of the protein, subsequently altering the carrier's phenotype. This protocol describes the use of the 'Sorting Tolerant From Intolerant' (SIFT) algorithm in predicting whether an AAS affects protein function. To assess the effect of a substitution, SIFT assumes that important positions in a protein sequence have been conserved throughout evolution and therefore substitutions at these positions may affect protein function. Thus, by using sequence homology, SIFT predicts the effects of all possible substitutions at each position in the protein sequence. The protocol typically takes 5-20 min, depending on the input. SIFT is available as an online tool (http://sift.jcvi.org). PMID- 19561591 TI - MicroRNA-mediated switching of chromatin-remodelling complexes in neural development. AB - One of the most distinctive steps in the development of the vertebrate nervous system occurs at mitotic exit when cells lose multipotency and begin to develop stable connections that will persist for a lifetime. This transition is accompanied by a switch in ATP-dependent chromatin-remodelling mechanisms that appears to coincide with the final mitotic division of neurons. This switch involves the exchange of the BAF53a (also known as ACTL6a) and BAF45a (PHF10) subunits within Swi/Snf-like neural-progenitor-specific BAF (npBAF) complexes for the homologous BAF53b (ACTL6b) and BAF45b (DPF1) subunits within neuron-specific BAF (nBAF) complexes in post-mitotic neurons. The subunits of the npBAF complex are essential for neural-progenitor proliferation, and mice with reduced dosage for the genes encoding its subunits have defects in neural-tube closure similar to those in human spina bifida, one of the most serious congenital birth defects. In contrast, BAF53b and the nBAF complex are essential for an evolutionarily conserved program of post-mitotic neural development and dendritic morphogenesis. Here we show that this essential transition is mediated by repression of BAF53a by miR-9* and miR-124. We find that BAF53a repression is mediated by sequences in the 3' untranslated region corresponding to the recognition sites for miR-9* and miR-124, which are selectively expressed in post-mitotic neurons. Mutation of these sites led to persistent expression of BAF53a and defective activity dependent dendritic outgrowth in neurons. In addition, overexpression of miR-9* and miR-124 in neural progenitors caused reduced proliferation. Previous studies have indicated that miR-9* and miR-124 are repressed by the repressor-element-1 silencing transcription factor (REST, also known as NRSF). Indeed, expression of REST in post-mitotic neurons led to derepression of BAF53a, indicating that REST mediated repression of microRNAs directs the essential switch of chromatin regulatory complexes. PMID- 19561592 TI - Demonstration of two-qubit algorithms with a superconducting quantum processor. AB - Quantum computers, which harness the superposition and entanglement of physical states, could outperform their classical counterparts in solving problems with technological impact-such as factoring large numbers and searching databases. A quantum processor executes algorithms by applying a programmable sequence of gates to an initialized register of qubits, which coherently evolves into a final state containing the result of the computation. Building a quantum processor is challenging because of the need to meet simultaneously requirements that are in conflict: state preparation, long coherence times, universal gate operations and qubit readout. Processors based on a few qubits have been demonstrated using nuclear magnetic resonance, cold ion trap and optical systems, but a solid-state realization has remained an outstanding challenge. Here we demonstrate a two qubit superconducting processor and the implementation of the Grover search and Deutsch-Jozsa quantum algorithms. We use a two-qubit interaction, tunable in strength by two orders of magnitude on nanosecond timescales, which is mediated by a cavity bus in a circuit quantum electrodynamics architecture. This interaction allows the generation of highly entangled states with concurrence up to 94 per cent. Although this processor constitutes an important step in quantum computing with integrated circuits, continuing efforts to increase qubit coherence times, gate performance and register size will be required to fulfil the promise of a scalable technology. PMID- 19561593 TI - Regulation of the innate immune response by threonine-phosphatase of Eyes absent. AB - Innate immunity is stimulated not only by viral or bacterial components, but also by non-microbial danger signals (damage-associated molecular patterns). One of the damage-associated molecular patterns is chromosomal DNA that escapes degradation. In programmed cell death and erythropoiesis, DNA from dead cells or nuclei expelled from erythroblasts is digested by DNase II in the macrophages after they are engulfed. DNase II(-/-) (also known as Dnase2a(-/-)) mice suffer from severe anaemia or chronic arthritis due to interferon-beta (IFN-beta) and tumour necrosis factor-alpha (TNF-alpha) produced from the macrophages carrying undigested DNA in a Toll-like receptor (TLR)-independent mechanism. Here we show that Eyes absent 4 (EYA4), originally identified as a co-transcription factor, stimulates the expression of IFN-beta and CXCL10 in response to the undigested DNA of apoptotic cells. EYA4 enhanced the innate immune response against viruses (Newcastle disease virus and vesicular stomatitis virus), and could associate with signalling molecules (IPS-1 (also known as MAVS), STING (TMEM173) and NLRX1). Three groups have previously shown that EYA has phosphatase activity. We found that mouse EYA family members act as a phosphatase for both phosphotyrosine and phosphothreonine. The haloacid dehalogenase domain at the carboxy terminus contained the tyrosine-phosphatase, and the amino-terminal half carried the threonine-phosphatase. Mutations of the threonine-phosphatase, but not the tyrosine-phosphatase, abolished the ability of EYA4 to enhance the innate immune response, suggesting that EYA regulates the innate immune response by modulating the phosphorylation state of signal transducers for the intracellular pathogens. PMID- 19561594 TI - Rapid and systematic analysis of the RNA recognition specificities of RNA-binding proteins. AB - Metazoan genomes encode hundreds of RNA-binding proteins (RBPs) but RNA-binding preferences for relatively few RBPs have been well defined. Current techniques for determining RNA targets, including in vitro selection and RNA co immunoprecipitation, require significant time and labor investment. Here we introduce RNAcompete, a method for the systematic analysis of RNA binding specificities that uses a single binding reaction to determine the relative preferences of RBPs for short RNAs that contain a complete range of k-mers in structured and unstructured RNA contexts. We tested RNAcompete by analyzing nine diverse RBPs (HuR, Vts1, FUSIP1, PTB, U1A, SF2/ASF, SLM2, RBM4 and YB1). RNAcompete identified expected and previously unknown RNA binding preferences. Using in vitro and in vivo binding data, we demonstrate that preferences for individual 7-mers identified by RNAcompete are a more accurate representation of binding activity than are conventional motif models. We anticipate that RNAcompete will be a valuable tool for the study of RNA-protein interactions. PMID- 19561597 TI - Periodic rotation of magnetization in a non-centrosymmetric soft magnet induced by an electric field. AB - The control of magnetism with an electric field is a challenging area with the potential to affect fields related to magnetic data storage, sensors and magnetic random access memory. Although there are some successful examples of such control based on the use of magnetic metals and semiconductors, energy loss caused by current flow is a problem that needs to be addressed. In particular, the repeatable control of magnetization with an electric field can be disturbed by joule heat loss. In this regard, non-centrosymmetric insulating magnets are good candidates for controlling magnetization without energy loss, in which the linear magnetoelectric effect has an essential role. Moreover, such magnets exhibit an unconventional magneto-optical effect, which allows the time-resolved detection of the magnetization direction. Here, we show a periodic oscillation of the magnetization direction by +/-20 degrees in a non-centrosymmetric soft magnet (Cu,Ni)B(2)O(4), which is induced by an a.c. electric field of 2 kHz. The present study provides a strategy for identifying materials in which the magnetization direction can be modulated at high speed with an electric field. PMID- 19561595 TI - Sequential treatment of drug-resistant tumors with targeted minicells containing siRNA or a cytotoxic drug. AB - The dose-limiting toxicity of chemotherapeutics, heterogeneity and drug resistance of cancer cells, and difficulties of targeted delivery to tumors all pose daunting challenges to effective cancer therapy. We report that small interfering RNA (siRNA) duplexes readily penetrate intact bacterially derived minicells previously shown to cause tumor stabilization and regression when packaged with chemotherapeutics. When targeted via antibodies to tumor-cell surface receptors, minicells can specifically and sequentially deliver to tumor xenografts first siRNAs or short hairpin RNA (shRNA)-encoding plasmids to compromise drug resistance by knocking down a multidrug resistance protein. Subsequent administration of targeted minicells containing cytotoxic drugs eliminate formerly drug-resistant tumors. The two waves of treatment, involving minicells loaded with both types of payload, enable complete survival without toxicity in mice with tumor xenografts, while involving several thousandfold less drug, siRNA and antibody than needed for conventional systemic administration of cancer therapies. PMID- 19561596 TI - Multi-site assessment of the precision and reproducibility of multiple reaction monitoring-based measurements of proteins in plasma. AB - Verification of candidate biomarkers relies upon specific, quantitative assays optimized for selective detection of target proteins, and is increasingly viewed as a critical step in the discovery pipeline that bridges unbiased biomarker discovery to preclinical validation. Although individual laboratories have demonstrated that multiple reaction monitoring (MRM) coupled with isotope dilution mass spectrometry can quantify candidate protein biomarkers in plasma, reproducibility and transferability of these assays between laboratories have not been demonstrated. We describe a multilaboratory study to assess reproducibility, recovery, linear dynamic range and limits of detection and quantification of multiplexed, MRM-based assays, conducted by NCI-CPTAC. Using common materials and standardized protocols, we demonstrate that these assays can be highly reproducible within and across laboratories and instrument platforms, and are sensitive to low mug/ml protein concentrations in unfractionated plasma. We provide data and benchmarks against which individual laboratories can compare their performance and evaluate new technologies for biomarker verification in plasma. PMID- 19561598 TI - An omnidirectional retroreflector based on the transmutation of dielectric singularities. AB - Transformation optics is a concept used in some metamaterials to guide light on a predetermined path. In this approach, the materials implement coordinate transformations on electromagnetic waves to create the illusion that the waves are propagating through a virtual space. Transforming space by appropriately designed materials makes devices possible that have been deemed impossible. In particular, transformation optics has led to the demonstration of invisibility cloaking for microwaves, surface plasmons and infrared light. Here, on the basis of transformation optics, we implement a microwave device that would normally require a dielectric singularity, an infinity in the refractive index. To fabricate such a device, we transmute a dielectric singularity in virtual space into a mere topological defect in a real metamaterial. In particular, we demonstrate an omnidirectional retroreflector, a device for faithfully reflecting images and for creating high visibility from all directions. Our method is robust, potentially broadband and could also be applied to visible light using similar techniques. PMID- 19561599 TI - Printed artificial cilia from liquid-crystal network actuators modularly driven by light. AB - Polymeric microactuators are potentially useful in micromechanical systems and lab-on-a-chip systems. However, manufacturing of miniature polymeric actuators has been complicated owing to the necessity of including electrodes for actuation or using lithographic techniques for patterning. Here, we demonstrate that all polymer microdevices can be fabricated using inkjet printing technology in combination with self-organizing liquid-crystal network actuators. We exploit the self-assembling properties of the liquid crystal to create large strain gradients, and light-driven actuation is chosen to allow simple and remote addressing. By using multiple inks, microactuators with different subunits are created that can be selectively addressed by changing the wavelength of the light. The actuators mimic the motion of natural cilia. These artificial cilia have the potential to create flow and mixing in wet environments such as lab-on-a chip applications. The process is easily adapted for roll-to-roll fabrication, allowing for large-scale and low-cost production of miniaturized active polymer systems. PMID- 19561600 TI - Selective positioning of organic dyes in a mesoporous inorganic oxide film. AB - Although sequential adsorption of dyes in a single TiO(2) electrode is ideal to extend the range of light absorption in dye-sensitized solar cells, high temperature processing has so far limited its application. We report a method for selective positioning of organic dye molecules with different absorption ranges in a mesoporous TiO(2) film by mimicking the concept of the stationary phase and the mobile phase in column chromatography, where polystyrene-filled mesoporous TiO(2) film is explored for use as a stationary phase and a Bronsted-base containing polymer solution is developed for use as a mobile phase for selective desorption of the adsorbed dye. By controlling the desorption and adsorption depth, yellow, red and green dyes were vertically aligned within a TiO(2) film, which is confirmed by an electron probe micro-analyser. The external quantum efficiency (EQE) spectrum from a solar cell with three selectively positioned dyes reveals the EQE characteristics of each single-dye cell. PMID- 19561601 TI - A discrete alcohol pocket involved in GIRK channel activation. AB - Ethanol modifies neural activity in the brain by modulating ion channels. Ethanol activates G protein-gated inwardly rectifying K(+) channels, but the molecular mechanism is not well understood. Here, we used a crystal structure of a mouse inward rectifier containing a bound alcohol and structure-based mutagenesis to probe a putative alcohol-binding pocket located in the cytoplasmic domains of GIRK channels. Substitutions with bulkier side-chains in the alcohol-binding pocket reduced or eliminated activation by alcohols. By contrast, alcohols inhibited constitutively open channels, such as IRK1 or GIRK2 engineered to strongly bind PIP(2). Mutations in the hydrophobic alcohol-binding pocket of these channels had no effect on alcohol-dependent inhibition, suggesting an alternate site is involved in inhibition. Comparison of high-resolution structures of inwardly rectifying K(+) channels suggests a model for activation of GIRK channels using this hydrophobic alcohol-binding pocket. These results provide a tool for developing therapeutic compounds that could mitigate the effects of alcohol. PMID- 19561602 TI - Resolving single cone inputs to visual receptive fields. AB - With the current techniques available for mapping receptive fields, it is impossible to resolve the contribution of single cone photoreceptors to the response of central visual neurons. Using adaptive optics to correct for ocular aberrations, we delivered micron-scale spots of light to the receptive field centers of neurons in the macaque lateral geniculate nucleus. Parvocellular LGN neurons mapped in this manner responded with high reliability to stimulation of single cones. PMID- 19561603 TI - A trophic role for Wnt-Ror kinase signaling during developmental pruning in Caenorhabditis elegans. AB - The molecular mechanism by which neurites are selected for elimination or incorporation into the mature circuit during developmental pruning remains unknown. The trophic theory postulates that local cues provided by target or surrounding cells act to inhibit neurite elimination. However, no widely conserved factor mediating this trophic function has been identified. We found that the developmental survival of specific neurites in Caenorhabditis elegans largely depends on detection of the morphogen Wnt by the Ror kinase CAM-1, which is a transmembrane tyrosine kinase with a Frizzled domain. Mutations in Wnt genes or in cam-1 enhanced neurite elimination, whereas overexpression of cam-1 inhibited neurite elimination in a Wnt-dependent manner. Moreover, mutations in these genes counteracted the effect of a mutation in mbr-1, which encodes a transcription factor that promotes neurite elimination. These results reveal the trophic role of an atypical Wnt pathway and reinforce the classical model of developmental pruning. PMID- 19561604 TI - The common colorectal cancer predisposition SNP rs6983267 at chromosome 8q24 confers potential to enhanced Wnt signaling. AB - Homozygosity for the G allele of rs6983267 at 8q24 increases colorectal cancer (CRC) risk approximately 1.5 fold. We report here that the risk allele G shows copy number increase during CRC development. Our computer algorithm, Enhancer Element Locator (EEL), identified an enhancer element that contains rs6983267. The element drove expression of a reporter gene in a pattern that is consistent with regulation by the key CRC pathway Wnt. rs6983267 affects a binding site for the Wnt-regulated transcription factor TCF4, with the risk allele G showing stronger binding in vitro and in vivo. Genome-wide ChIP assay revealed the element as the strongest TCF4 binding site within 1 Mb of MYC. An unambiguous correlation between rs6983267 genotype and MYC expression was not detected, and additional work is required to scrutinize all possible targets of the enhancer. Our work provides evidence that the common CRC predisposition associated with 8q24 arises from enhanced responsiveness to Wnt signaling. PMID- 19561605 TI - Mutations affecting the secretory COPII coat component SEC23B cause congenital dyserythropoietic anemia type II. AB - Congenital dyserythropoietic anemias (CDAs) are phenotypically and genotypically heterogeneous diseases. CDA type II (CDAII) is the most frequent CDA. It is characterized by ineffective erythropoiesis and by the presence of bi- and multinucleated erythroblasts in bone marrow, with nuclei of equal size and DNA content, suggesting a cytokinesis disturbance. Other features of the peripheral red blood cells are protein and lipid dysglycosylation and endoplasmic reticulum double-membrane remnants. Development of other hematopoietic lineages is normal. Individuals with CDAII show progressive splenomegaly, gallstones and iron overload potentially with liver cirrhosis or cardiac failure. Here we show that the gene encoding the secretory COPII component SEC23B is mutated in CDAII. Short hairpin RNA (shRNA)-mediated suppression of SEC23B expression recapitulates the cytokinesis defect. Knockdown of zebrafish sec23b also leads to aberrant erythrocyte development. Our results provide in vivo evidence for SEC23B selectivity in erythroid differentiation and show that SEC23A and SEC23B, although highly related paralogous secretory COPII components, are nonredundant in erythrocyte maturation. PMID- 19561606 TI - Sequence variants in the CLDN14 gene associate with kidney stones and bone mineral density. AB - Kidney stone disease is a common condition. To search for sequence variants conferring risk of kidney stones, we conducted a genome-wide association study in 3,773 cases and 42,510 controls from Iceland and The Netherlands. We discovered common, synonymous variants in the CLDN14 gene that associate with kidney stones (OR = 1.25 and P = 4.0 x 10(-12) for rs219780[C]). Approximately 62% of the general population is homozygous for rs219780[C] and is estimated to have 1.64 times greater risk of developing the disease compared to noncarriers. The CLDN14 gene is expressed in the kidney and regulates paracellular permeability at epithelial tight junctions. The same variants were also found to associate with reduced bone mineral density at the hip (P = 0.00039) and spine (P = 0.0077). PMID- 19561607 TI - The 8q24 cancer risk variant rs6983267 shows long-range interaction with MYC in colorectal cancer. AB - An inherited variant on chromosome 8q24, rs6983267, is significantly associated with cancer pathogenesis. We present evidence that the region harboring this variant is a transcriptional enhancer, that the alleles of rs6983267 differentially bind transcription factor 7-like 2 (TCF7L2) and that the risk region physically interacts with the MYC proto-oncogene. These data provide strong support for a biological mechanism underlying this non-protein-coding risk variant. PMID- 19561608 TI - Structural determinants of gating in the TRPV1 channel. AB - Transient receptor potential vanilloid 1 (TRPV1) channels mediate several types of physiological responses. Despite the importance of these channels in pain detection and inflammation, little is known about how their structural components convert different types of stimuli into channel activity. To localize the activation gate of these channels, we inserted cysteines along the S6 segment of mutant TRPV1 channels and assessed their accessibility to thiol-modifying agents. We show that access to the pore of TRPV1 is gated by S6 in response to both capsaicin binding and increases in temperature, that the pore-forming S6 segments are helical structures and that two constrictions are present in the pore: one that impedes the access of large molecules and the other that hampers the access of smaller ions and constitutes an activation gate of these channels. PMID- 19561609 TI - The structure of SHH in complex with HHIP reveals a recognition role for the Shh pseudo active site in signaling. AB - Hedgehog (Hh) signaling is crucial for many aspects of embryonic development, whereas dysregulation of this pathway is associated with several types of cancer. Hedgehog-interacting protein (Hhip) is a surface receptor antagonist that is equipotent against all three mammalian Hh homologs. The crystal structures of human HHIP alone and bound to Sonic hedgehog (SHH) now reveal that HHIP is comprised of two EGF domains and a six-bladed beta-propeller domain. In the complex structure, a critical loop from HHIP binds the pseudo active site groove of SHH and directly coordinates its Zn2+ cation. Notably, sequence comparisons of this SHH binding loop with the Hh receptor Patched (Ptc1) ectodomains and HHIP- and PTC1-peptide binding studies suggest a 'patch for Patched' at the Shh pseudo active site; thus, we propose a role for Hhip as a structural decoy receptor for vertebrate Hh. PMID- 19561610 TI - H3K64 trimethylation marks heterochromatin and is dynamically remodeled during developmental reprogramming. AB - Histone modifications are central to the regulation of all DNA-dependent processes. Lys64 of histone H3 (H3K64) lies within the globular domain at a structurally important position. We identify trimethylation of H3K64 (H3K64me3) as a modification that is enriched at pericentric heterochromatin and associated with repeat sequences and transcriptionally inactive genomic regions. We show that this new mark is dynamic during the two main epigenetic reprogramming events in mammals. In primordial germ cells, H3K64me3 is present at the time of specification, but it disappears transiently during reprogramming. In early mouse embryos, it is inherited exclusively maternally; subsequently, the modification is rapidly removed, suggesting an important role for H3K64me3 turnover in development. Taken together, our findings establish H3K64me3 as a previously uncharacterized histone modification that is preferentially localized to repressive chromatin. We hypothesize that H3K64me3 helps to 'secure' nucleosomes, and perhaps the surrounding chromatin, in an appropriately repressed state during development. PMID- 19561611 TI - Structural insights into hedgehog ligand sequestration by the human hedgehog interacting protein HHIP. AB - Hedgehog (Hh) morphogens have fundamental roles in development, whereas dysregulation of Hh signaling leads to disease. Multiple cell-surface receptors are responsible for transducing and/or regulating Hh signals. Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. Zn2+ makes a key contribution to the Hh-HHIP interface, whereas Ca2+ is likely to prevent electrostatic repulsion between the two proteins, suggesting an important modulatory role. This interplay of several metal binding sites suggests a tuneable mechanism for regulation of Hh signaling. PMID- 19561612 TI - Mycobacterium tuberculosis evades macrophage defenses by inhibiting plasma membrane repair. AB - Induction of macrophage necrosis is a strategy used by virulent Mycobacterium tuberculosis (Mtb) to avoid innate host defense. In contrast, attenuated Mtb causes apoptosis, which limits bacterial replication and promotes T cell cross priming by antigen-presenting cells. Here we show that Mtb infection causes plasma membrane microdisruptions. Resealing of these lesions, a process crucial for preventing necrosis and promoting apoptosis, required translocation of lysosomal and Golgi apparatus-derived vesicles to the plasma membrane. Plasma membrane repair depended on prostaglandin E(2) (PGE(2)), which regulates synaptotagmin 7 (Syt-7), the calcium sensor involved in the lysosome-mediated repair mechanism. By inducing production of lipoxin A(4) (LXA(4)), which blocks PGE(2) biosynthesis, virulent Mtb prevented membrane repair and induced necrosis. Thus, virulent Mtb impairs macrophage plasma membrane repair to evade host defenses. PMID- 19561613 TI - Essential function for the GTPase TC21 in homeostatic antigen receptor signaling. AB - T cell antigen receptors (TCRs) and B cell antigen receptors (BCRs) transmit low grade signals necessary for the survival and maintenance of mature cell pools. We show here that TC21, a small GTPase encoded by Rras2, interacted constitutively with both kinds of receptors. Expression of a dominant negative TC21 mutant in T cells produced a rapid decrease in cell viability, and Rras2(-/-) mice were lymphopenic, possibly as a result of diminished homeostatic proliferation and impaired T cell and B cell survival. In contrast, TC21 was overexpressed in several human lymphoid malignancies. Finally, the p110delta catalytic subunit of phosphatidylinositol-3-OH kinase (PI(3)K) was recruited to the TCR and BCR in a TC21-dependent way. Consequently, we propose TC21 directly links antigen receptors to PI(3)K-mediated survival pathways. PMID- 19561615 TI - Mina, an Il4 repressor, controls T helper type 2 bias. AB - T helper type 2 (T(H)2) bias, which is the propensity of naive CD4(+) T cells to differentiate into interleukin 4 (IL-4)-secreting T(H)2 cells, is a genetic trait that affects susceptibility to infectious, autoimmune and allergic diseases. T(H)2 bias correlates with the amount of IL-4 initially secreted by newly activated helper T cells that feeds back positively through the pathway of the IL 4 receptor and the transcription factors STAT6 and GATA-3 to drive T(H)2 development. Here we identify Mina, a member of the jumonji C (JmjC) protein family, as a genetic determinant of T(H)2 bias. Mina specifically bound to and repressed the Il4 promoter. Mina overexpression in transgenic mice impaired Il4 expression, whereas its knockdown in primary CD4(+) T cells led to Il4 derepression. Our findings collectively provide mechanistic insight into an Il4 regulatory pathway that controls helper T cell differentiation and genetic variation in T(H)2 bias. PMID- 19561614 TI - Immunoglobulin D enhances immune surveillance by activating antimicrobial, proinflammatory and B cell-stimulating programs in basophils. AB - Immunoglobulin D (IgD) is an enigmatic antibody isotype that mature B cells express together with IgM through alternative RNA splicing. Here we report active T cell-dependent and T cell-independent IgM-to-IgD class switching in B cells of the human upper respiratory mucosa. This process required activation-induced cytidine deaminase (AID) and generated local and circulating IgD-producing plasmablasts reactive to respiratory bacteria. Circulating IgD bound to basophils through a calcium-mobilizing receptor that induced antimicrobial, opsonizing, inflammatory and B cell-stimulating factors, including cathelicidin, interleukin 1 (IL-1), IL-4 and B cell-activating factor (BAFF), after IgD crosslinking. By showing dysregulation of IgD class-switched B cells and 'IgD-armed' basophils in autoinflammatory syndromes with periodic fever, our data indicate that IgD orchestrates an ancestral surveillance system at the interface between immunity and inflammation. PMID- 19561616 TI - Activation of kinin receptor B1 limits encephalitogenic T lymphocyte recruitment to the central nervous system. AB - Previous proteomic and transcriptional analyses of multiple sclerosis lesions revealed modulation of the renin-angiotensin and the opposing kallikrein-kinin pathways. Here we identify kinin receptor B1 (Bdkrb1) as a specific modulator of immune cell entry into the central nervous system (CNS). We demonstrate that the Bdkrb1 agonist R838 (Sar-[D-Phe]des-Arg(9)-bradykinin) markedly decreases the clinical symptoms of experimental autoimmune encephalomyelitis (EAE) in SJL mice, whereas the Bdkrb1 antagonist R715 (Ac-Lys-[D-betaNal(7), Ile(8)]des-Arg(9) bradykinin) resulted in earlier onset and greater severity of the disease. Bdkrb1 deficient (Bdkrb1(-/-)) C57BL/6 mice immunized with a myelin oligodendrocyte glycoprotein fragment, MOG(35-55), showed more severe disease with enhanced CNS immune cell infiltration. The same held true for mixed bone marrow-chimeric mice reconstituted with Bdkrb1(-/-) T lymphocytes, which showed enhanced T helper type 17 (T(H)17) cell invasion into the CNS. Pharmacological modulation of Bdkrb1 revealed that in vitro migration of human T(H)17 lymphocytes across blood-brain barrier endothelium is regulated by this receptor. Taken together, these results suggest that the kallikrein-kinin system is involved in the regulation of CNS inflammation, limiting encephalitogenic T lymphocyte infiltration into the CNS, and provide evidence that Bdkrb1 could be a new target for the treatment of chronic inflammatory diseases such as multiple sclerosis. PMID- 19561617 TI - Tenascin-C is an endogenous activator of Toll-like receptor 4 that is essential for maintaining inflammation in arthritic joint disease. AB - Although there have been major advances in the treatment of rheumatoid arthritis with the advent of biological agents, the mechanisms that drive cytokine production and sustain disease chronicity remain unknown. Tenascin-C (encoded by Tnc) is an extracellular matrix glycoprotein specifically expressed at areas of inflammation and tissue damage in inflamed rheumatoid joints. Here we show that mice that do not express tenascin-C show rapid resolution of acute joint inflammation and are protected from erosive arthritis. Intra-articular injection of tenascin-C promotes joint inflammation in vivo in mice, and addition of exogenous tenascin-C induces cytokine synthesis in explant cultures from inflamed synovia of individuals with rheumatoid arthritis. Moreover, in human macrophages and fibroblasts from synovia of individuals with rheumatoid arthritis, tenascin-C induces synthesis of proinflammatory cytokines via activation of Toll-like receptor 4 (TLR4). Thus, we have identified tenascin-C as a novel endogenous activator of TLR4-mediated immunity that mediates persistent synovial inflammation and tissue destruction in arthritic joint disease. PMID- 19561619 TI - Bioactivity-guided mapping and navigation of chemical space. AB - The structure- and chemistry-based hierarchical organization of library scaffolds in tree-like arrangements provides a valid, intuitive means to map and navigate chemical space. We demonstrate that scaffold trees built using bioactivity as the key selection criterion for structural simplification during tree construction allow efficient and intuitive mapping, visualization and navigation of the chemical space defined by a given library, which in turn allows correlation of this chemical space with the investigated bioactivity and further compound design. Brachiation along the branches of such trees from structurally complex to simple scaffolds with retained yet varying bioactivity is feasible at high frequency for the five major pharmaceutically relevant target classes and allows for the identification of new inhibitor types for a given target. We provide proof of principle by identifying new active scaffolds for 5-lipoxygenase and the estrogen receptor ERalpha. PMID- 19561618 TI - Targeted depletion of lymphotoxin-alpha-expressing TH1 and TH17 cells inhibits autoimmune disease. AB - Uncontrolled T helper type 1 (T(H)1) and T(H)17 cells are associated with autoimmune responses. We identify surface lymphotoxin-alpha (LT-alpha) as common to T(H)0, T(H)1 and T(H)17 cells and employ a unique strategy to target these subsets using a depleting monoclonal antibody (mAb) directed to surface LT-alpha. Depleting LT-alpha-specific mAb inhibited T cell-mediated models of delayed-type hypersensitivity and experimental autoimmune encephalomyelitis. In collagen induced arthritis (CIA), preventive and therapeutic administration of LT-alpha specific mAb inhibited disease, and immunoablated T cells expressing interleukin 17 (IL-17), interferon-gamma and tumor necrosis factor-alpha (TNF-alpha), whereas decoy lymphotoxin-beta receptor (LT-betaR) fusion protein had no effect. A mutation in the Fc tail, rendering the antibody incapable of Fcgamma receptor binding and antibody-dependent cellular cytotoxicity activity, abolished all in vivo effects. Efficacy in CIA was preceded by a loss of rheumatoid-associated cytokines IL-6, IL-1beta and TNF-alpha within joints. These data indicate that depleting LT-alpha-expressing lymphocytes with LT-alpha-specific mAb may be beneficial in the treatment of autoimmune disease. PMID- 19561620 TI - Interactive exploration of chemical space with Scaffold Hunter. AB - We describe Scaffold Hunter, a highly interactive computer-based tool for navigation in chemical space that fosters intuitive recognition of complex structural relationships associated with bioactivity. The program reads compound structures and bioactivity data, generates compound scaffolds, correlates them in a hierarchical tree-like arrangement, and annotates them with bioactivity. Brachiation along tree branches from structurally complex to simple scaffolds allows identification of new ligand types. We provide proof of concept for pyruvate kinase. PMID- 19561623 TI - There is no such thing as a tree of life (and of course viruses are out!). PMID- 19561621 TI - Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli. AB - Absolute metabolite concentrations are critical to a quantitative understanding of cellular metabolism, as concentrations impact both the free energies and rates of metabolic reactions. Here we use LC-MS/MS to quantify more than 100 metabolite concentrations in aerobic, exponentially growing Escherichia coli with glucose, glycerol or acetate as the carbon source. The total observed intracellular metabolite pool was approximately 300 mM. A small number of metabolites dominate the metabolome on a molar basis, with glutamate being the most abundant. Metabolite concentration exceeds K(m) for most substrate-enzyme pairs. An exception is lower glycolysis, where concentrations of intermediates are near the K(m) of their consuming enzymes and all reactions are near equilibrium. This may facilitate efficient flux reversibility given thermodynamic and osmotic constraints. The data and analyses presented here highlight the ability to identify organizing metabolic principles from systems-level absolute metabolite concentration data. PMID- 19561624 TI - Compelling reasons why viruses are relevant for the origin of cells. PMID- 19561625 TI - Viral genomes are part of the phylogenetic tree of life. PMID- 19561626 TI - Ten good reasons not to exclude giruses from the evolutionary picture. PMID- 19561627 TI - Six comments on the ten reasons for the demotion of viruses. PMID- 19561628 TI - Reasons to include viruses in the tree of life. PMID- 19561629 TI - The most important news at ASCO 2009. PMID- 19561630 TI - Stereotactic radiosurgery alone to treat brain metastases. PMID- 19561631 TI - Cetuximab, chemotherapy and KRAS status in mCRC. PMID- 19561632 TI - ASCT in follicular lymphoma. PMID- 19561633 TI - Role of lymphadenectomy in the staging of endometrial cancer. PMID- 19561634 TI - Cervical cancer in rural India. PMID- 19561635 TI - Apples and oranges: the low and mid versus the upper rectum. PMID- 19561636 TI - A stepwise approach to the management of metastatic midgut carcinoid tumor. AB - BACKGROUND: A 48-year-old man presented with diarrhea, flushing, abdominal pain and weight loss of 10 kg over a 6-month period. He subsequently developed dyspnea on exertion. INVESTIGATIONS: Physical examination, laboratory tests, CT of the abdomen, liver biopsy, echocardiography, immunohistochemistry staining of the biopsy specimen for neuroendocrine markers including chromogranin A, synaptophysin and protein gene product 9.5, and (111)In-pentetreotide scintigraphy (Octreoscan). DIAGNOSIS: Carcinoid tumor of midgut origin with large segment 3 liver metastasis. Carcinoid syndrome and carcinoid heart disease. MANAGEMENT: Symptomatic relief with somatostatin analog therapy and subsequent resection of the segment 3 liver metastasis. Tricuspid and pulmonary valve replacement. PMID- 19561637 TI - Stereological study of the effects of vitamin E on testis structure in rats treated with para-nonylphenol. AB - This study was organized to see whether vitamin E, as a strong antioxidant, could affect the abnormalities of testis structure caused by para-nonylphenol (p-NP) during its development. A total of 32 female Wistar rats after mating were divided into four groups (n = 8): control, vitamin E (100 mg kg(-1) per day), p NP (250 mg kg(-1) per day) and p-NP + vitamin E. The rats were treated from the seventh day of pregnancy till the twenty-first day. After weaning, the male pups were divided into the same groups and were treated orally for 90 days. Finally, the right testis was fixed, processed, stained and studied using stereological methods. The weight and volume of testis, volume of seminiferous tubules and its diameter, thickness of the basement membrane, height of the germinal epithelium, total number of types A and B spermatogonia, spermatocyte, spermatid and Sertoli cells were significantly reduced in p-NP group when compared with other groups. Co-administration of vitamin E and p-NP compensated for the adverse effects of p NP on the above parameters. In addition, treatment with only vitamin E caused a significant increase in diameter, basement membrane thickness and height of germinal epithelium, number of spermatogonia and spermatocytes. Co-administration of vitamin E with p-NP could prevent the adverse effects of p-NP on the testis structure during its development. PMID- 19561639 TI - SOCS1 protects protein tyrosine phosphatases by thioredoxin upregulation and attenuates Jaks to suppress ROS-mediated apoptosis. AB - Suppressors of cytokine signaling (SOCS) are negative regulators of cytokine induced signal transduction, which play multiple roles in cell growth, differentiation and apoptosis. In this study, the regulatory role of SOCS in oxidative stress-induced apoptosis was investigated. In Jurkat T cells and mouse splenocytes, we have found that SOCS1 is induced in response to tumor necrosis factor-alpha or H(2)O(2), concomitant with the activation of Jaks which act as important mediators of reactive oxygen species (ROS)-induced apoptosis upstream of p38 mitogen-activated protein kinase. Using SOCS1 overexpressing or knockdown Jurkat T-cell systems we clearly demonstrate that, SOCS1 inhibits the ROS mediated apoptosis. The antiapoptotic action of SOCS1 was exerted not only by suppressing Jaks, but also by sustaining protein tyrosine phosphatase (PTP) activities. Notably, SOCS1-transduced cells displayed increase in thioredoxin levels and decrease in ROS generation induced by oxidative stress. In addition, the Jak-inhibiting and PTP-sustaining effect of SOCS1 was significantly reduced on thioredoxin ablation. Moreover, coimmunoprecipitation data revealed molecular interaction of SHP1 or CD45 with thioredoxin, which was promoted in SOCS1 transfected cells. Together, our data strongly suggest that both the protection of PTPs by thioredoxin from ROS attack and the attenuation of Jaks account for the antiapoptotic function of SOCS1 in immune cells under oxidative stress. PMID- 19561638 TI - Nucleophosmin protein expression level, but not threonine 198 phosphorylation, is essential in growth and proliferation. AB - Nucleophosmin (NPM), an oligomeric phosphoprotein and nucleolar target of the ARF tumor suppressor, contributes to several critical cellular processes. Previous studies have shown that the human NPM's phosphorylation by cyclin E-cyclin dependent kinase 2 (cdk2) on threonine (Thr) 199 regulates its translocation from the centrosome during cell cycle progression. Given our previous finding that ARF directly binds NPM, impeding its transit to the cytoplasm and arresting cells before S-phase entry, we hypothesized that ARF might also inhibit NPM phosphorylation. However, ARF induction did not impair phosphorylation of the cdk2 target residue in murine NPM, Thr198. Furthermore, phosphorylation of Thr198 occurred throughout the cell cycle and was concomitant with increases in overall NPM expression. To investigate the cell's presumed requirement for NPM-Thr198 phosphorylation in promoting the processes of growth and proliferation, we examined the effects of a non-phosphorylatable NPM mutant, T198A, in a clean cell system in which endogenous NPM had been removed by RNA interference. Here, we show that the T198A mutant is fully capable of executing NPM's described roles in nucleocytoplasmic shuttling, ribosome export and cell cycle progression. Moreover, the proliferative defects observed with stable NPM knockdown were restored by mutant NPM-T198A expression. Thus, we demonstrate that the reduction in NPM protein expression blocks cellular growth and proliferation, whereas phosphorylation of NPM-Thr198 is not essential for NPM's capacity to drive cell cycle progression and proliferation. PMID- 19561640 TI - WW domain containing E3 ubiquitin protein ligase 1 targets the full-length ErbB4 for ubiquitin-mediated degradation in breast cancer. AB - ErbB4, a member of the epidermal growth factor receptor family, plays a role in normal breast and breast cancer development by regulating mammary epithelial cell proliferation, survival and differentiation. In this study, we show that WWP1, a C2-WW-HECT type E3 ubiquitin ligase, binds, ubiquitinates and destructs ErbB4 CYT1, but much less efficiently for CYT2, isoforms (both JMa and JMb). The protein-protein interaction occurs primarily between the first and third WW domains of WWP1 and the second PY motif of ErbB4. Knockdown of WWP1 by two different small interfering RNAs increases the endogenous ErbB4 protein levels in both MCF7 and T47D breast cancer cell lines. In addition, overexpression of the wild type, but not the catalytic inactive WWP1, dramatically decreases the endogenous ErbB4 protein levels in MCF7. Importantly, we found that WWP1 negatively regulates the heregulin-beta1-stimulated ErbB4 activity as measured by the serum response element report assay and the BRCA1 mRNA expression. After a systematic screening of all WWP1 family members by small interfering RNA, we found that AIP4/Itch and HECW1/NEDL1 also negatively regulate the ErbB4 protein expression in T47D. Interestingly, the protein expression levels of both WWP1 and ErbB4 are higher in estrogen receptor-alpha-positive than in estrogen receptor alpha-negative breast cancer cell lines. These data suggest that WWP1 and its family members suppress the ErbB4 expression and function in breast cancer. PMID- 19561641 TI - Post-translational modification and stability of low molecular weight cyclin E. AB - Our laboratory has previously described the presence of five tumor-specific low molecular weight isoforms of cyclin E in both tumor cell lines and breast cancer patient biopsies. We have also shown that one of these low forms arises from an alternate start site, whereas the other four appear as two sets of doublets following cleavage through an elastase-like enzyme. However, the origin of both sets of doublets was unknown. Here, we demonstrate that the larger isoform of each doublet is the result of phosphorylation at a key degradation site. Through site-directed mutagenesis of different phosphorylation sites within the cyclin E protein, we discovered that phosphorylation of threonine 395 is responsible for generating the larger isoform of each doublet. Because phosphorylation of threonine 395 has been linked to the proteasome-mediated degradation of full length cyclin E, we examined the stability of T395A phospho-mutants in both non tumorigenic mammary epithelial cells and tumor cells. The results revealed that the low molecular weight isoforms appear to be stable in both a tumor cell line and a non-tumor forming cell line regardless of the presence of this critical phosphorylation site. The stability of low molecular weight cyclin E may have implications for both tumorigenesis and treatment of tumors expressing them. PMID- 19561642 TI - Involvement of mitochondria and recruitment of Fas/CD95 signaling in lipid rafts in resveratrol-mediated antimyeloma and antileukemia actions. AB - We have found that resveratrol (trans-3,4',5-trihydroxystilbene) induced apoptosis in multiple myeloma (MM) and T-cell leukemia cells through coclustering of Fas/CD95 death receptor and lipid rafts, whereas normal lymphocytes were spared. Tumor necrosis factor-related apoptosis-inducing ligand receptors, Fas associated death domain-containing protein (FADD), procaspase-8, procaspase-10, c Jun amino-terminal kinase and Bid were also recruited into lipid rafts on resveratrol incubation with MM and T-cell leukemia cells. Raft disruption inhibited resveratrol-induced apoptosis. Bcl-XL overexpression prevented resveratrol-induced disruption of mitochondrial transmembrane potential (DeltaPsi(m)) and apoptosis. A FADD dominant-negative mutant, that blocked Fas/CD95 downstream signaling, precluded resveratrol-induced DeltaPsi(m) loss and apoptosis, indicating a sequence of Fas/CD95 signaling-->mitochondrion in the apoptotic response triggered by resveratrol. Cells deficient in Fas/CD95 did not undergo resveratrol-induced apoptosis. Pretreatment of MM cells with interferon gamma upregulated Fas/CD95 and caspase-8, and potentiated resveratrol-induced apoptosis. Our data indicate that recruitment of Fas/CD95 death receptor and downstream signaling molecules into lipid rafts, followed by DeltaPsi(m) disruption, underlies the apoptotic action of resveratrol in MM and T-cell leukemic cells. Combination of resveratrol with perifosine or bortezomib potentiated the apoptotic response induced by each single drug. These results also highlight the role of recruitment of Fas/CD95 signaling in lipid rafts in antimyeloma and antileukemia chemotherapy. PMID- 19561643 TI - Autonomous regulation of osteosarcoma cell invasiveness by Wnt5a/Ror2 signaling. AB - The receptor tyrosine kinase Ror2 regulates cell migration by acting as a receptor or co-receptor for Wnt5a. Although Wnt5a has been implicated in the invasiveness of several types of tumors, the role of Ror2 in tumor invasion remains elusive. Here we show that osteosarcoma cell lines SaOS-2 and U2OS show invasive properties in vitro by activating Wnt5a/Ror2 signaling in a cell autonomous manner. The suppressed expression of either Wnt5a or Ror2 in osteosarcoma cells inhibits cell invasiveness accompanying decreased invadopodia formation. Gene-expression profiling identified matrix metalloproteinase 13 (MMP 13) as one of the genes whose expression is downregulated in SaOS-2 cells following suppression of Ror2 expression. Reduced expression or activity of MMP 13 suppresses invasiveness of SaOS-2 cells. Moreover, expression of MMP-13 and cell invasiveness by Wnt5a/Ror2 signaling can be abrogated by an inhibitor of the Src-family protein tyrosine kinases (SFKs), suggesting the role of the SFKs in MMP-13 expression through Wnt5a/Ror2 signaling. We further show that activation of an SFK is inhibited by the suppressed expression of Ror2. Collectively, these results indicate that Wnt5a/Ror2 signaling involves the activation of a SFK, leading to MMP-13 expression, and that constitutively active Wnt5a/Ror2 signaling confers invasive properties on osteosarcoma cells in a cell-autonomous manner. PMID- 19561645 TI - Mammalian cell-cycle regulation: several Cdks, numerous cyclins and diverse compensatory mechanisms. AB - After a decade of extensive work on gene knockout mouse models of cell-cycle regulators, the classical model of cell-cycle regulation was seriously challenged. Several unexpected compensatory mechanisms were uncovered among cyclins and Cdks in these studies. The most astonishing observation is that Cdk2 is dispensable for the regulation of the mitotic cell cycle with both Cdk4 and Cdk1 covering for Cdk2's functions. Similar to yeast, it was recently discovered that Cdk1 alone can drive the mammalian cell cycle, indicating that the regulation of the mammalian cell cycle is highly conserved. Nevertheless, cell cycle-independent functions of Cdks and cyclins such as in DNA damage repair are still under investigation. Here we review the compensatory mechanisms among major cyclins and Cdks in mammalian cell-cycle regulation. PMID- 19561644 TI - Accelerated mammary maturation and differentiation, and delayed MMTVneu-induced tumorigenesis of K303R mutant ERalpha transgenic mice. AB - We identified a somatic mutation in estrogen receptor-alpha (ERalpha) in breast cancer causing a lysine to arginine transition (K303R) resulting in hypersensitivity to estrogen, altered associations with coactivators and corepressors and altered posttranslational modifications of ERalpha. We have developed a transgenic mouse expressing the K303R mutant ERalpha under control of the mouse mammary tumor virus (MMTV) promoter. At 4 months of age, K303R ERalpha transgenic animals demonstrate precocious alveolar budding compared with wild type ERalpha transgenic mice or nontransgenic littermates. Despite these morphologic differences, K303R ERalpha transgenic mice displayed no differences in levels of ERalpha, progesterone receptor or proliferation at this time-point. Pregnancy or chronic estrogen plus progesterone exposure in K303R ERalpha transgenic mice also resulted in significantly more alveolar budding, increased beta-casein production and dilated ducts when compared with nontransgenic littermates. To examine the effects of mutant expression on tumorigenesis, mutant ERalpha mice were crossed with FVB-MMTVneu mice and significantly delayed time to neu-mediated tumorigenesis in bigenic animals. In contrast, mutant expression did not affect carcinogen-induced tumorigenesis. Collectively, these data demonstrate that aberrant estrogenic signaling through the K303R ERalpha mutation may lead to precocious alveolar budding in virgin mice, and to an expedited maturation and differentiation phenotype in the mammary glands of hormonally stimulated animals. PMID- 19561647 TI - Cytotoxic crossmatch analysis before allo-SCT is a poor diagnostic tool for prediction of rejection. AB - The predictive value of cytotoxic crossmatch analysis before allo-SCT remains unclear. We retrospectively analyzed the clinical impact of cytotoxic T- and B cell crossmatch testing before allo-SCT between January 2000 and June 2005. Cytotoxic crossmatches were performed in 157 patients receiving stem cells from matched unrelated donors or an HLA-A, -B or -DRB1 allele mismatched graft. Ninety patients are still alive. Eleven patients rejected their grafts. One of 11 patients with rejection was positive in a T-cell crossmatch before allo-SCT and 4 of 11 in B-cell crossmatches. T-cell crossmatches showed a sensitivity of 9% and a specificity of 97% compared with 36 and 86% for B-cell crossmatches. Positive T and/or B-crossmatch before SCT had no predictive value for survival in this study as compared with patients with a negative crossmatch. In conclusion, the pretransplant cytotoxic T- and/or B-crossmatch is a poor predictor of rejection after allo-SCT. PMID- 19561646 TI - Functional RET G691S polymorphism in cutaneous malignant melanoma. AB - RET proto-oncogene encodes a receptor tyrosine kinase whose ligand is glial cell line-derived neurotrophic factor (GDNF), and its polymorphism at G691S juxtamembrane region (RETp) is a germline polymorphism. Cutaneous melanomas, particularly the desmoplastic subtype, are highly neurotropic; thus we sought to determine the frequency of RETp in cutaneous melanoma and its functional responsiveness to GDNF. RETp was assessed in 71 non-desmoplastic cutaneous melanomas (non-DMs) and 70 desmoplastic melanomas (DMs). Melanoma cell lines with RETp, RET wild type (RETwt), BRAF V600E mutation (BRAFmt) or BRAF wild type (BRAFwt) were assessed for functional activity. RETp frequency was significantly higher in DMs (61%) than in non-DMs (31%, P<0.001). BRAFmt was detected in only 11% of DMs. GDNF stimulation significantly amplified cell proliferation, migration and invasion in RETp, but not in RETwt melanoma cells. GDNF stimulation of RETp cell lines enhanced phosphorylation of extracellular signal-regulated kinase (ERK) and Akt of the RET-RAS-RAF-ERK and RET-phosphatidylinositol 3-kinase (PI3K)-Akt pathways, respectively. GDNF response of RETp cells in signal transduction and other functional studies were not affected by BRAFmt. The study demonstrates that RETp is frequently found in cutaneous melanoma, particularly desmoplastic subtypes, and responds to GDNF inducing events favorable for tumor progression. PMID- 19561648 TI - Infections in non-myeloablative hematopoietic stem cell transplantation patients with lymphoid malignancies: spectrum of infections, predictors of outcome and proposed guidelines for fungal infection prevention. AB - The overall risk of infections is lower in patients undergoing non-myeloablative allogeneic stem cell transplantation (NST) than in conventional stem cell transplant recipients. We sought to evaluate conditions associated with increased risk of infections after NST. In 81 patients, 187 infection episodes were noted; chronic lymphocytic leukemia (138 episodes/100 person-years) and recipients of matched unrelated donor graft (128 episodes/100 person-years) had higher risk of infection. Only half of the cytomegalovirus (CMV) infections occurred 31-100 days after transplantation. Most patients with CMV infection were non-neutropenic (100%), had lymphoma (76%), were younger (<55 years; 72%) and had received matched related donor (MRD) graft (72%). However, graft-versus-host disease (GVHD) was present in only 15% of these patients. Seven (78%) of nine invasive fungal infections (IFI) were diagnosed >100 days after NST and were associated with high mortality (78%). Most patients with IFI were also not neutropenic (100%), had received MRD graft (100%), had lymphoma (78%) and were given systemic steroids (78%); unlike CMV infection, 67% of these patients also had GVHD. On the basis of our results, we propose that NST recipients with lymphoma treated with high-dose corticosteroids for GVHD be considered for antifungal prophylaxis or pre-emptive antifungal therapy. PMID- 19561650 TI - Dasatinib may not suppress the GVL effect of donor lymphocyte infusions for CML. PMID- 19561651 TI - Double allogeneic hematopoietic SCT as a rescue therapy for poor-risk hematological malignancies. AB - The prognosis of patients with poor-risk or relapsed hematological malignancies is dismal. The dose intensification necessary to achieve subsequent CR is limited by the toxicity of chemotherapy. Treatment intensification with double allogeneic HSCT (dHSCT) may enhance the antileukemic effect and reduces treatment-related toxicity associated with prolonged aplasia during reinduction. We evaluated this approach in 23 patients, nine with primary refractory disease or relapse after conventional chemotherapy (group I) and 14 with relapses after allogeneic HSCT (group II). Double HSCT was feasible in all patients. At the end of the observation period, 6 of 23 (26%) patients were still alive and in remission with a median observation time of 60 months (1-153). The overall survival probability at 1 year was 41% (95% confidence interval (CI), 21-62%), transplant-related mortality (TRM) 28% (9-47%) and the incidence of relapse 42% (18-66%). The TRM in groups I and II were 22 and 36% and the relapse rate 33 and 50%, respectively. In conclusion, we have shown the feasibility of dHSCT with an acceptable TRM, irrespective of a previous allogeneic HSCT. Whether this approach offers a survival benefit for patients with poor-risk leukemias has to be tested in larger prospective trials. PMID- 19561652 TI - [Free-floating literature]. PMID- 19561649 TI - Influence of oral beclomethasone dipropionate on early non-infectious pulmonary outcomes after allogeneic hematopoietic cell transplantation: results from two randomized trials. AB - Early non-infectious pulmonary complications represent a significant cause of mortality after hematopoietic cell transplantation (HCT). We tested the hypothesis that oral beclomethasone dipropionate (BDP) is effective for preventing early non-infectious pulmonary complications after allogeneic HCT. We retrospectively reviewed the medical records of 120 patients, 60 in each treatment arm, to identify non-infectious and infectious pulmonary events and pulmonary function test results from all patients who participated in two randomized trials of oral BDP for treatment of acute gastrointestinal GVHD. 17 Beclomethasone monopropionate (17-BMP), the active metabolite of BDP, was evaluated in blood from the right atrium in four patients. Thirty-three of 42 (79%) placebo-treated patients experienced a decrease of the DL(CO) from pretransplant to day 80 after transplant, compared with 27 of 49 (55%) BDP treated patients (P=0.02). In the first 200 days after randomization, there were no cases of non-infectious pulmonary complications in BDP-treated patients, vs four cases among placebo-treated patients (P=0.04). Levels of 17-BMP were detected in atrial blood at steady state. Delivery of a potent glucocorticoid such as 17-BMP to the pulmonary artery after oral dosing of BDP may be useful in modulating pulmonary inflammation and preventing the development of non infectious pulmonary complications after allogeneic HCT. PMID- 19561653 TI - [Health and medical research law]. PMID- 19561654 TI - [Cesarean section--for benefit or damage?]. PMID- 19561655 TI - [Between Scylla and Charybdis]. PMID- 19561656 TI - [General practioners and patients' general life problems]. AB - BACKGROUND: Many contact general practitioners (GPs) about non-medical problems in their lives; a situation which may be frustrating for GPs. MATERIAL AND METHODS: Semi-structured in-depth interviews with six Norwegian GPs by e-mail. The interviews were analysed according to a discourse analysis of which role the GPs considered they had, whether they saw themselves as therapists and how they perceived the personal strain related to working in general medicine. RESULTS: The majority of GPs did not distinguish clearly between somatic and more general life-related problems. Nevertheless, they defined medical problems as quite strictly somatic and saw themselves mainly as experts in somatic medicine. They described themselves more as spiritual advisers (pastor-like) than as psychotherapists. Some meant that their work had given them more self- confidence and made them more tolerant, but many said they felt helpless in their role as doctors. INTERPRETATION: GPs are doctors for all people. They have to relate to healthy and sick individuals with various medical and general life problems, and often experience a discrepancy between their medical competence and the challenges they meet. PMID- 19561657 TI - [Risk of infections among orienteers]. AB - BACKGROUND: Research on orienteers is useful for assessing the risk of infections associated with physical activity in the forest. In this paper four types of infections are reviewed, and the efficacy of preventive initiatives is discussed. MATERIAL AND METHODS: The paper is based on literature retrieved from a non systemic search in PubMed. RESULTS: Hepatitis B infection was more prevalent among orienteers before they were obliged to use protective clothing. In the 1980s, there was an increase of sudden unexpected death among young Swedish orienteers. Bartonella infection was later suggested as an underlying cause. No unexpected deaths have occurred among young orienteers after 1992 when specific advice was given regarding training and competitions. Orienteers do not seem to be affected by lyme borreliosis or tick-borne encephalitis (TBE) more often than others, but only two old studies have been performed. INTERPRETATION: Orienteers may be at risk of acquiring infection from lyme borreliosis and TBE in Norway in the future, as the incidence of these contagions is increasing. Norwegian medical personnel should consider TBE vaccination of orienteers and others who wander in areas with a high prevalence of infected ticks. PMID- 19561658 TI - [Increased rate of Caesarean sections--causes and consequences]. AB - BACKGROUND: A rise in the rate of caesarean sections has been observed in most parts of the developed world during the last decades. Causes and consequences are much debated. MATERIAL AND METHODS: Non-systematic literature search in PubMed. RESULTS: The increased rate of caesarean sections can be explained by both medical non-medical factors. Among the medical factors are increases in maternal age and body mass index, as well as changes in obstetric practise and technology. Some non-medical factors are caesarean section requested by the mother, fear of litigation among caregivers and inappropriate organization of maternity care. Caesarean section is associated with maternal postpartum morbidity, reduced fertility and placental complications in a subsequent pregnancy. For the child, caesarean section is associated with postpartum respiratory morbidity, less breast-feeding and possibly more atopic disease. For society, caesarean section is more costly than vaginal delivery. INTERPRETATION: There are many and complex causes of the rise in caesarean section rates in industrialized countries. The procedure has inherent negative consequences (short- and long-term) for mother and child, as well as being an economic burden to society. There is every reason to attempt prevention of a further increase in caesarean section rates. PMID- 19561659 TI - [New international guidelines on antithrombotic therapy in atrial fibrillation]. AB - BACKGROUND: The American College of Cardiology/American Heart Association/European Society of Cardiology and American College of Chest Physicians have recently revised international guidelines on antithrombotic therapy in atrial fibrillation. This may influence clinical practice in Norway. MATERIAL AND METHODS: Potential impact on Norwegian clinical practice is discussed in light of the guidelines mentioned above and other relevant literature. RESULTS: Several studies have indicated that the risk of stroke associated with atrial fibrillation is lower than previously anticipated, and the revised international guidelines have taken this into account. The new guidelines emphasize the CHADS2 score as a tool to decide which patients should receive warfarin. Points are assigned based on simple clinical characteristics of the patients: Congestive heart failure 1 point; Hypertension 1 point; Age > 75 years 1 point; Diabetes 1 point; Prior stroke or transitory ischemic attack 2 points. Aspirin is recommended to patients with a CHADS2 score of 0 points; warfarin or aspirin to patients with 1 point, and warfarin to patients with >or= 2 points. INTERPRETATION: The new international guidelines give a slightly higher threshold for recommending warfarin to patients with atrial fibrillation. The CHADS2 score, which is based on simple clinical characteristics, has been shown to be reliable and may contribute to improved risk stratification in patients with atrial fibrillation. PMID- 19561660 TI - [Medication errors in hospitalised patients]. AB - BACKGROUND: Medication errors can arise both during prescription and administration (dispensing and distribution) of drugs. Little is known about types of medication errors in Norwegian hospitals. MATERIAL AND METHOD: All medication errors reported at St. Olav's Hospital from 1 July 2002 to 30 June 2006 were reviewed and analysed. RESULTS: 610 reports were identified. The most common cause of reporting (39 %) was prescription of a different dose from the one prescribed. Other frequent causes were administration of a different drug than the one prescribed (17 %), inadvertent subcutaneous infusion of an intravenous drug (15 %), and that the drug was given to another patient (12 %). The errors were almost exclusively reported by nurses. In 107 cases (18 %), precautions had been taken to reduce the extent of injury after the error had been identified. The causes of errors could be classified in three main categories: Nonvigilance caused by stress, lack of appropriate routines or violation of them, and lack of appropriate skills/negligence. INTERPRETATION: Changes of routines, improved education in existing routines, and increased pharmacological competence may contribute to prevention of medication errors. PMID- 19561661 TI - [Oral antibiotic treatment of urinary tract infections in children]. AB - Urinary tract infection (UTI) is one of the most common bacterial infections in childhood. Empiric antibiotic therapy is guided by the clinical presentation, the patient's ability to take oral agents and the local resistance pattern of Escherichia coli (E. coli), the most common pathogen. Most children (with both upper and lower UTI) can safely be treated with oral antibiotics. We recommend pivmecillinam or amoxicillin-clavulanic acid as first-line empiric therapy for upper UTI. Amoxicillin and trimethoprim-sulfamethoxazole are no longer first-line oral agents due to increasing E. coli resistance to both drugs. For lower UTI nitrofurantoin is an excellent first choice. In Norway, lack of paediatric antibiotic syrups is a great challenge and we recommend that such formulations are introduced to the Norwegian market. PMID- 19561662 TI - [Informed choice--important also for Norwegian pregnant women]. PMID- 19561663 TI - [Jellyfish sting injuries]. PMID- 19561664 TI - [Heart arrest--definition and occurrence]. PMID- 19561665 TI - [Fine needle aspiration cytology--complement to diagnostics of hypothyroidism]. PMID- 19561666 TI - [Incomplete survey of the Epley maneuver]. PMID- 19561668 TI - [The dialogue between general practitioners and specialists care must be improved!]. PMID- 19561669 TI - [Compensation claims following meningococcal vaccine B trial]. PMID- 19561671 TI - [Elderly physicians and authorization]. PMID- 19561670 TI - [Unfounded accusations against the Norwegian Registration Authority for Health Personnel]. PMID- 19561672 TI - [Individual primary prevention of cardiovascular diseases]. PMID- 19561673 TI - [Children of drug addicts must get a broader care]. PMID- 19561675 TI - [Determination of disability compensation--a difficult matter]. PMID- 19561676 TI - [Drugs against pollen allergy, pregnancy and breast feeding]. PMID- 19561677 TI - [Difficult--but important--with critical assessment of new technology]. PMID- 19561678 TI - [Avalanche research in Norway]. PMID- 19561683 TI - [Nutritional situation for mothers and children in South Africa]. AB - BACKGROUND: Even 15 years after the downfall of the apartheid regime, there are significant inequalities regarding resources, welfare and social benefits among the people of South Africa. Poverty prevails and conditions are bad with respect to sanitation, health and living conditions; a situation which is likely to affect nutritional health. We have reviewed the nutritional status in South Africa. MATERIAL AND METHODS: This article is based on literature retrieved from non-systematic reviews of the databases PubMed and High Wire Press, in addition to information from official documents and African journals. The author's recent field work in South Africa was also a source of information. RESULTS: Nationwide surveys, as well as smaller studies have documented high rates of stunting, malnutrition and deficiency of micronutrients among South African children. Daily hunger is reported from more than half of South African households. There is also a rise in the number of overweight and obese children and mothers. HIV/aids is highly prevalent and often coexists with tuberculosis. Many of these patients have a poor nutritional status, which in turn reduces the effect of antiviral treatment. INTERPRETATION: The high rate of malnutrition and HIV/aids among mothers and children in South Africa will delay improvement of general health in this population with generations. PMID- 19561685 TI - The Birth of the Western Journal of Emergency Medicine: WestJEM. PMID- 19561686 TI - Asthma: Effect of genotype on Response to Therapy in the Emergency Department. AB - OBJECTIVE: We examined the effect of two beta2-adrenoreceptor (beta2AR) polymorphisms (A46G and C79G) in asthmatics presenting to the Emergency Department (ED) in relation to their response to standard therapy measured by change in Forced Expiratory Volume at one second (FEV1). Our hypothesis was that the polymorphisms in the beta2AR gene would predict clinical response to therapy with 46G and 79C displaying decreased response to inhaled therapy. METHODS: This was a pilot feasibility study of a convenience sample of patients seen in the ED for acute exacerbation of asthma. Baseline data collected included: age, gender, ethnicity, vital signs, baseline FEV1, body mass index (BMI), smoking history and medications taken prior to arrival to the ED. Patients received standard ED care and FEV1 was measured after each treatment. Blood was taken and genotyped. RESULTS: Fifty-three patients were enrolled over a three-month period. Using mean improvement in FEV1 from baseline to the first treatment as the primary outcome of interest, we performed multivariable linear regression analyses, with the FEV1 change as the dependent variable. When modeled as an ordinal covariate representing the number of G alleles present, there was a significant positive trend for the C79G locus (p=0.035). Those who were GG homozygotes had a 0.284 L/min improvement in FEV1 (31%) after their initial albuterol treatment compared to 0.123 L/min (12%) in those who were CC homozygotes. This represents a 2.5 times relative difference and a 19% actual difference. Genotypes at the A46G locus were not associated with FEV1 change. CONCLUSION: In this pilot study of ED patients with acute asthma exacerbation, there was a significant effect of genotype on response to therapy. PMID- 19561687 TI - The Holy grail, genes and Number Needed to Treat. PMID- 19561688 TI - A Prospective, Randomized Trial in the Emergency Department of Suggestive Audio Therapy under Deep Sedation for Smoking Cessation. AB - OBJECTIVES: In a sample of patients undergoing procedural deep sedation in the emergency department (ED), we conducted a prospective, randomized, single-blinded trial of audio-therapy for smoking cessation. METHODS: We asked subjects about their smoking, including desire to quit (0-10 numerical scale) and number of cigarettes smoked per day. Subjects were randomized to either a control tape (music alone) or a tape with repeated smoking-cessation messages over music. Tapes were started with first doses of sedation and stopped with patient arousal. Telephone follow-up occurred between two weeks and three months to assess the number of cigarettes smoked per day. Study endpoints were self-reported complete cessation and decrease of half or more in total cigarettes smoked per day. RESULTS: One hundred eleven patients were enrolled in the study, 54 to intervention and 57 to control. Mean desire to quit was 7.15 +/- 2.6 and mean cigarettes per day was 17.5 +/- 12.1. We successfully contacted 69 (62%) patients. Twenty-seven percent of intervention and 26% of control patients quit (mean difference = 1%; 95% CI: -22.0% to 18.8%). Thirty-seven percent of intervention and 51% of control patients decreased smoking by half or more (mean difference = 14.6%; 95% CI: -8.7% to 35.6%). CONCLUSION: Suggestive audio-therapy delivered during deep sedation in the ED did not significantly decrease self reported smoking behavior. PMID- 19561689 TI - Educational assessment of medical student rotation in emergency ultrasound. AB - BACKGROUND: Medical student ultrasound education is sparse. In 2002, we began the first medical student rotation in emergency ultrasound. OBJECTIVE: To evaluate if medical students can learn and retain sonographic skills during a two- or four week elective. METHODS: We gave students an exam on the first and last days of the rotation. Six months later, students took the exam a third time. A control group was used for comparison. RESULTS: Over a 19-month period, we enrolled 45 students (25 on the two-week and 20 on the four-week elective). The four-week student post-test score was significantly better than the two- week post-test score (81% vs 72%, p=0.003). On the six-month exam, the four-week student post test score was significantly better than the two-week post-test score (77% vs 69%, p=0.008). The control group did not statistically improve. CONCLUSION: Medical students can learn bedside ultrasound interpretation with clinical integration and retain the knowledge six months later. PMID- 19561691 TI - Recovery from Severe Hyperthermia (45 degrees C) and Rhabdomyolysis Induced by Methamphetamine Body-Stuffing. PMID- 19561690 TI - The basics of alcohol screening, brief intervention and referral to treatment in the emergency department. AB - Nearly eight million emergency department (ED) visits are attributed to alcohol every year in the United States. A substantial proportion is due to trauma. In 2005, 16,885 people were killed as a result of alcohol-related motor vehicle crashes. Patients with alcohol-use problems (AUPs) are not only more likely to drive after drinking but are also at greater risk for serious alcohol-related illness and injury. Emergency departments have an important and unique opportunity to identify these patients and intervene during the "teachable moment" of an ED visit. The American College of Emergency Physicians, Emergency Nurses Association, American College of Surgeons-Committee on Trauma, American Public Health Association, and the National Highway Traffic Safety Administration, have identified Alcohol Screening, Brief Intervention and Referral to Treatment (SBIRT) as a pivotal injury- and illness-prevention strategy to improve the health and well-being of ED patients. We provide a general overview of the basis and need for integrating SBIRT into EDs. Models of SBIRT, as well as benefits and challenges to its implementation, are also discussed. PMID- 19561692 TI - Images in Emergency Medicine: CSF Hydrothorax. PMID- 19561693 TI - New onset thyrotoxicosis presenting as vomiting, abdominal pain and transaminitis in the emergency department. AB - This case report describes an unusual presentation of an emergency department (ED) patient with nausea, vomiting, and epigastric pain, who was initially suspected of having viral hepatitis. The patient returned to the ED seven days later with persistent tachycardia and was diagnosed with new onset thyrotoxicosis. PMID- 19561694 TI - Accuracy of ED Bedside Ultrasound for Identification of gallstones: retrospective analysis of 575 studies. AB - STUDY OBJECTIVE: To determine the ability of emergency department (ED) physicians to diagnose cholelithiasis with bedside ultrasound. METHODS: ED gallbladder ultrasounds recorded over 37 months were compared to radiology ultrasound interpretation. RESULTS: Of 1,690 ED gallbladder ultrasound scans performed during this period, radiology ultrasound was performed in 575/1690 (34%) cases. ED physician bedside interpretation was 88% sensitive [95% CI, 84-91] and 87% specific [95% CI, 82-91], while positive predictive value (PPV) was 91% [88-94%] and negative predictive value (NPV) was 83% [78-87%], using radiology interpretation as the criterion reference. CONCLUSION: ED physician ultrasound of the gallbladder for cholelithiasis is both sensitive and specific. PMID- 19561695 TI - Validation of the Social Security Death Index (SSDI): An Important Readily Available Outcomes Database for Researchers. AB - STUDY OBJECTIVE: To determine the accuracy of the online Social Security Death Index (SSDI) for determining death outcomes. METHODS: We selected 30 patients who were determined to be dead and 90 patients thought to be alive after an ED visit as determined by a web-based searched of the SSDI. For those thought to be dead we requested death certificates. We then had a research coordinator blinded to the results of the SSDI search, complete direct follow-up by contacting the patients, family or primary care physicians to determine vital status. To determine the sensitivity and specificity of the SSDI for death at six months in this cohort, we used direct follow-up as the criterion reference and calculated 95% confidence intervals. RESULTS: Direct follow-up was completed for 90% (108 of 120) of the patients. For those patients 20 were determined to be dead and 88 alive. The dead were more likely to be male (57%) and older [(mean age 83.9 (95% CI 79.1 - 88.7) vs. 60.9 (95% CI 56.4 - 65.4) for those alive]. The sensitivity of the SSDI for those with completed direct follow-up was 100% (95% CI 91 -100%) with specificity of 100% (95% CI 98-100%). Of the 12 patients who were not able to be contacted through direct follow-up, the SSDI indicated that 10 were dead and two were alive. CONCLUSIONS: SSDI is an accurate measure of death outcomes and appears to have the advantage of finding deaths among patients lost to follow up. PMID- 19561696 TI - Under Utilization of local anesthetics in infant lumbar punctures. AB - BACKGROUND: Lumbar Puncture (LP) is an invasive procedure frequently used to diagnose meningitis among the pediatric population. Neonates and infants have not routinely received local anesthesia prior to LP. STUDY OBJECTIVE: To determine whether emergency medicine physicians and pediatricians use local analgesics on neonates and infants prior to performing an LP and to identify which local anesthetics, if any, were used. METHODS: Prospective, cohort study of all infants, six months of age or less, that received an LP in the emergency department (ED) or inpatient pediatric units for suspected meningitis during a period of year at a university tertiary care hospital. RESULTS: A total sample population of 111 infants that received an LP within the study period. A control population of 42 adults received an LP. Only 40.4% (45/111) of the infants received local analgesia prior to LP: either 1% lidocaine, EMLA or a combination of the two. Infants were less likely to receive lidocaine or EMLA prior to LP compared to adult subjects (OR= 0.27; 95% CI0.12 to 0.62). No neonates that were less than one month of age received local procedural anesthesia by emergency medicine or pediatric physicians. ED physicians' use of local anesthesia prior to LP increased with increasing age of the infant. The pediatricians in this study used local anesthesia prior to LP when the infant was at least five months of age. DISCUSSION: The data objectively support recent literature regarding the under use or lack of use of analgesia prior to LP among neonates and infants. Local anesthetics should be used routinely without exception prior to performing an LP in the pediatric population. PMID- 19561697 TI - Adaptation of Predictive Models to PDA Hand-Held Devices. AB - Prediction models using multiple logistic regression are appearing with increasing frequency in the medical literature. Problems associated with these models include the complexity of computations when applied in their pure form, and lack of availability at the bedside. Personal digital assistant (PDA) hand held devices equipped with spreadsheet software offer the clinician a readily available and easily applied means of applying predictive models at the bedside. The purposes of this article are to briefly review regression as a means of creating predictive models and to describe a method of choosing and adapting logistic regression models to emergency department (ED) clinical practice. PMID- 19561698 TI - Combining ketamine and propofol ("ketofol") for emergency department procedural sedation and analgesia: a review. PMID- 19561699 TI - Ten solutions for emergency department crowding. PMID- 19561701 TI - EKG Criteria for Fibrinolysis: What's Up with the J Point? PMID- 19561702 TI - Loss of digits after trauma in a patient with systemic lupus erythematosus. AB - A 19-year-old female with Systemic Lupus Erythematosus (SLE) presented with ischemia of her left hand following trauma. Medical therapy was initiated but failed to improve her symptoms, and revision amputation was ultimately performed. The patient's final diagnosis was digital ischemia due to secondary Raynaud's Phenomenon (RP). The authors discuss diagnosis, complications, and treatment of this relatively uncommon disorder. The authors report this case in order to discuss how secondary RP can be complicated by ischemia and the multidisciplinary approach that needs to take place to prevent the latter from occurring. PMID- 19561703 TI - Welcome to the Resident/Student/Fellow Section of WestJEM! PMID- 19561700 TI - Wide Complex Tachycardias: Understanding this Complex Condition: Part 1 - Epidemiology and Electrophysiology. PMID- 19561704 TI - Clinical Emergency Medicine Algorithms: Vaginal Bleeding in Early Pregnancy (Less than 20 weeks). PMID- 19561705 TI - Images in emergency medicine: aortic dissection. PMID- 19561706 TI - Images in emergency medicine: retinal detachment. PMID- 19561707 TI - Images in emergency medicine: retropharyngeal abscess. PMID- 19561708 TI - Images in emergency medicine : lisfranc fracture-dislocation. PMID- 19561709 TI - Interpretation of traumatic lumbar puncture in the setting of possible subarachnoid hemorrhage: who can be safely discharged? PMID- 19561710 TI - In reply. PMID- 19561711 TI - Comparison of the evaluations of a case-based reasoning decision support tool by specialist expert reviewers with those of end users. AB - BACKGROUND: Decision-support tools (DST) are typically developed by computer engineers for use by clinicians. Prototype testing DSTs may be performed relatively easily by one or two clinical experts. The costly alternative is to test each prototype on a larger number of diverse clinicians, based on the untested assumption that these evaluations would more accurately reflect those of actual end users. HYPOTHESIS: We hypothesized substantial or better agreement (as defined by a kappa statistic greater than 0.6) between the evaluations of a case based reasoning (CBR) DST predicting ED admission for bronchiolitis performed by the clinically diverse end users, to those of two clinical experts who evaluated the same DST output. METHODS: Three outputs from a previously described DST were evaluated by the emergency physicians (EP) who originally saw the patients and by two pediatric EPs with an interest in bronchiolitis. The DST outputs were as follows: predicted disposition, an example of another previously seen patient to explain the prediction, and explanatory dialog. Each was rated using the scale Definitely Not, No, Maybe, Yes, and Absolutely. This was converted to a Likert scale for analysis. Agreement was measured using the kappa statistic. RESULTS: Agreement with the DST predicted disposition was moderate between end users and the expert reviewers, but was only fair or poor for value of the explanatory case and dialog. CONCLUSION: Agreement between expert evaluators and end users on the value of a CBR DST predicted dispositions was moderate. For the more subjective explicative components, agreement was fair, poor, or worse. PMID- 19561712 TI - Leukocytosis as a predictor of severe injury in blunt trauma. AB - OBJECTIVE: The objective of this study was to determine if the white blood cell count can predict severity of injury in blunt trauma victims. METHODS: This was a retrospective study comparing two groups of blunt trauma victims by severity of injury, one with significant injury and one without significant injury, and comparing their initial WBC in the emergency department (ED). We also examined if WBC correlates with degree of injury using Injury Severity Score (ISS) in both groups combined. Further, we examined the WBC as a predictor of serious injury. RESULTS: Our study showed a difference in mean WBC between the two groups that was statistically significant (p<0.001). A positive relationship between ISS and WBC was found, although the association was weak (correlation coefficient = 0.369). While the WBC had moderate discriminatory capability for serious injury, it could not, in isolation, reliably rule in or out serious injury. Nevertheless, this study supports using WBC on presentation to the ED as an adjunct for making disposition decisions. CONCLUSION: A significant elevation in WBC in a blunt trauma patient, even with minimal initial signs of severe injury, should heighten suspicion for occult injury. PMID- 19561713 TI - Patient satisfaction and return to daily activities using etomidate procedural sedation for orthopedic injuries. AB - OBJECTIVES: With regard to sedative agents used in procedural sedation and analgesia (PSA), such as etomidate, the focus has been on variables usually related to side effect profile and the success rates of various procedures, with both variables specifically taking place during the patients' stay in the emergency department (ED). There have been no extensive data on the functional status of patients after they leave the ED following PSA. METHODS: Prospective questionnaire evaluating functional status among consecutive adult patients discharged from the ED after undergoing etomidate PSA for orthopedic procedures. RESULTS: The study sample consisted of 26 cases using only etomidate for closed orthopedic reductions. The mean age was 50.1 years (SD: 20.5), mean weight 86.3 kg (SD: 17.2), and 61.5% were males. The average dose of etomidate given was 0.14 mg/kg with 26.9% requiring a second dose of 0.11 mg/kg. The average dose of analgesic given was 0.11mg/kg in morphine equianalgesic units. The median time between procedural sedation and return to normal sleep was 36 hours, while return to operating a motor vehicle or return to work was 72 hours. Overall, 80% to 100% of respondents felt that any temporary dysfunction was secondary to the orthopedic problems and not to the procedural sedation. CONCLUSION: In this small follow-up study, adult patients undergoing PSA with etomidate for orthopedic closed reduction attribute post-discharge functional disability to the injury sustained and not to the PSA itself. PMID- 19561714 TI - The imminent healthcare and emergency care crisis in Japan. AB - OBJECTIVES: Japan has a universal healthcare system, and this paper describes the reality of the healthcare services provided, as well as current issues with the system. METHODS: Academic, government, and press reports on Japanese healthcare systems and healthcare guidelines were reviewed. RESULTS: The universal healthcare system of Japan is considered internationally to be both low-cost and effective because the Japanese population enjoys good health status with a long life expectancy, while healthcare spending in Japan is below the average given by the Organization for Economic Corporation and Development (OECD). However, in many regions of Japan the existing healthcare resources are seriously inadequate, especially with regard to the number of physicians and other health professionals. Because healthcare is traditionally viewed as "sacred" work in Japan, healthcare professionals are expected to make large personal sacrifices. Also, public attitudes toward medical malpractice have changed in recent decades, and medical professionals are facing legal issues without experienced support of the government or legal professionals. Administrative response to the lack of resources and collaboration among communities are beginning, and more efficient control and management of the healthcare system is under consideration. CONCLUSION: The Japanese healthcare system needs to adopt an efficient medical control organization to ease the strain on existing healthcare professionals and to increase the number of physicians and other healthcare resources. Rather than continuing to depend on healthcare professionals being able and willing to make personal sacrifices, the government, the public and medical societies must cooperate and support changes in the healthcare system. PMID- 19561715 TI - Wide Complex Tachycardias: Understanding this Complex Condition Part 2 - Management, Miscellaneous Causes, and Pitfalls. PMID- 19561716 TI - Takotsubo cardiomyopathy: a case series and review of the literature. AB - Takotsubo cardiomyopathy (TCM) is an unusual form of acute cardiomyopathy showing left ventricular apical ballooning. It is often triggered by intense physical or emotional distress. We report here four cases of TCM and a review of the literature on the topic. PMID- 19561717 TI - Chest swelling and Fever in an intravenous drug user. AB - This case report describes a sternoclavicular infection in an IV drug user. The history and physical exam suggested an abscess. In the emergency department (ED) the patient refused incision and drainage but did consent to simple needle aspiration. Subsequent culture of the aspirate revealed Pseudomonas aeruginosa. He was admitted for IV antibiotics. After admission, a bone scan suggested the presence of osteomyelitis. The patient refused operative debridement, but ultimately did consent to bedside incision and drainage. By day five, the fever had resolved and the patient signed out AMA. He was given a prescription for Ciprofloxacin. The patient had an unscheduled follow up in the ED five months later for an unrelated heroin overdose. Physical examination demonstrated complete resolution of the infection. PMID- 19561718 TI - Subchorionic hemorrhage appearing as twin gestation on endovaginal ultrasound. AB - This case study describes a pregnant patient with vaginal bleeding who had a bedside endovaginal ultrasound in the emergency department (ED). The emergency physician identified a live intra-uterine pregnancy (IUP) with another structure that appeared to be a second gestational sac. The patient subsequently had an endovaginal ultrasound in the radiology department 46 minutes later. The attending radiologist described one live IUP and a subchorionic hemorrhage. Comparison of the ED and radiology ultrasound showed that the second structure, identified as a subchorionic hemorrhage, had significantly decreased in size. Endovaginal ultrasound in the evaluation of possible ectopic pregnancy is a useful bedside tool in the ED. We discuss a pitfall that can occur with endocavitary ultrasound when a twin gestation is presumed. PMID- 19561719 TI - Point: diagnostic radiation: why aren't we stopping (or at least slowing down)? PMID- 19561720 TI - Counter-Point: are we really ordering too many CT scans? PMID- 19561721 TI - Images in emergency medicine: traumatic pneumocephalus. PMID- 19561722 TI - Images in emergency medicine: pediatric spinal cord injury without radiographic abnormality. PMID- 19561723 TI - Images in emergency medicine: left atrial myxoma. PMID- 19561724 TI - Images in emergency medicine: pacemaker extrusion causing chest pain. PMID- 19561725 TI - Images in emergency medicine: dermatomyositis. PMID- 19561727 TI - Health Literacy among Parents of Pediatric Patients. AB - BACKGROUND: Health literacy is an important predictor of healthcare outcomes, but research on this topic has largely been absent from the emergency medicine literature. OBJECTIVE: We measured the prevalence of health literacy in parents or guardians of pediatric patients seen in the emergency department (ED). METHODS: This was an observational study conducted in a Midwestern urban, university-based, tertiary, Level 1 trauma center ED with 33,000 visits/year. Using convenience sampling during a three-month period, English-speaking parents or guardians of pediatric patients (< 19 yrs.) were asked to complete the short version of the Test of Functional Health Literacy for Adults (s-TOFHLA). Parents/guardians were excluded if they had uncorrected visual impairment, required an interpreter, had altered mental status, or if the patients they accompanied were the subjects of a medical or trauma activation. RESULTS: Of the 188 parents or guardians approached, six did not consent or withdrew, one was excluded, leaving 181 (96.3%) in the study. Of these, 19 (10.5%) had either "marginal" or "inadequate" health literacy, while 162 (89.5%, 95% CI: 84.1%, 93.6%) had "adequate" health literacy. CONCLUSION: A large majority (89.5%) of English-speaking parents or guardians of pediatric patients evaluated in the ED have adequate health literacy. This data may prompt ED professionals to adjust their communication styles in the evaluation of children. Future multi-center studies are needed to confirm the findings in this pilot study. PMID- 19561728 TI - Prevalence of Bordetella pertussis and Bordetella parapertussis in Samples Submitted for RSV Screening. AB - BACKGROUND: The clinical presentation of Bordetella pertussis can overlap with that of respiratory syncytial virus (RSV); however, management differs. HYPOTHESIS: First, the prevalence of B. pertussis is less than 2% among patients screened for RSV, and second the prevalence of B. parapertussis is also less than 2% among these patients. METHODS: Nasal washings submitted to a clinical laboratory for RSV screening were tested for B. pertussis and B. parapertussis, using species-specific real-time polymerase chain reaction (PCR) assays. These were optimized to target conserved regions within a complement gene and the CarB gene, respectively. A Bordetella spp. genus-specific real-time PCR assay was designed to detect the Bhur gene of B. pertussis, B. parapertussis, and B. bronchiseptica. RSV A and B subtypes were tested by reverse transcription-PCR. RESULTS: Four hundred and eighty-nine clinical samples were tested. There was insufficient material to complete testing for one B. pertussis, 10 RSV subtype A, and four RSV subtype B assays. Bordetella pertussis was detected in 3/488 (0.6%) (95% CI 0.1% to 1.8%), while B. parapertussis was detected in 5/489 (1.0%) (95% CI 0.3% to 2.4%). Dual infection of B. pertussis with RSV and of B. parapertussis with RSV occurred in two and in three cases respectively. RSV was detected by PCR in 127 (26.5%). CONCLUSION: The prevalence of B. pertussis in nasal washings submitted for RSV screening was less than 2%. The prevalence of parapertussis may be higher than 2%. RSV with B. pertussis and RSV with B. parapertussis coinfection do occur. PMID- 19561729 TI - Self-reported emergency medicine residency applicant attitudes towards a procedural cadaver laboratory curriculum. AB - OBJECTIVE: Residency applicants consider a variety of factors when ranking emergency medicine (EM) programs for their NRMP match list. A human cadaver emergency procedure lab curriculum is uncommon. We hypothesized that the presence this curriculum would positively impact the ranking of an EM residency program. METHODS: The EM residency at Nebraska Medical Center is an urban, university based program with a PGY I-III format. Residency applicants during the interview for a position in the PGY I class of 2006 were surveyed by three weekly electronic mailings. The survey was distributed in March 2006 after the final NRMP match results were released. The survey explored learner preferences and methodological commonality of models of emergency procedural training, as well as the impact of a procedural cadaver lab curriculum on residency ranking. ANOVA of ranks was used to compare responses to ranking questions. RESULTS: Of the 73 potential subjects, 54 (74%) completed the survey. Respondents ranked methods of procedural instruction from 1 (most preferred or most common technique) to 4 (least preferred or least common technique). Response averages and 95% confidence intervals for the preferred means of learning a new procedure are as follows: textbook (3.69; 3.51-3.87), mannequin (2.83; 2.64-3.02), human cadaver (1.93; 1.72-2.14), and living patient (1.56; 1.33-1.79). Response averages for the commonality of means used to teach a new procedure are as follows: human cadaver (3.63; 3.46-3.80), mannequin (2.70; 2.50-2.90), living patient (2.09; 1.85-2.33), and textbook (1.57; 1.32-1.82). When asked if the University of Nebraska Medical Center residency ranked higher in the individual's match list because of its procedural cadaver lab, 14.8% strongly disagreed, 14.8% disagreed, 40.7% were neutral, 14.8% agreed, and 14.8% strongly agreed. CONCLUSION: We conclude that, although cadaveric procedural training is viewed by senior medical student learners as a desirable means of learning a procedure, its use is uncommon during medical school, and its presence as part of a residency curriculum does not influence ranking of the residency program. PMID- 19561730 TI - Tinnitus as a measure of salicylate toxicity in the overdose setting. AB - INTRODUCTION: The development of tinnitus and/or hearing loss (THL) in patients receiving chronic salicylate therapy has been demonstrated. However, to date, little scientific data validates this relationship in the large single overdose setting. OBJECTIVE: To correlate salicylate levels in patients with the subjective complaint of THL, following an acute salicylate overdose. METHODS: A retrospective chart review of cases of acute salicylate toxicity and THL reported to the Illinois Poison Control Center (IPC) from 2001-2002 was performed. Data abstracted included age, gender, ingestion time, salicylate levels, and arterial blood gases. RESULTS: Ninety-nine cases of THL were reviewed and analyzed with mean age of 23.7 years (SD: 10.9), 30.3% male, and 82.2% intentional overdoses. The average dose ingested was 20.0 grams (SD:20.2) and the mean time from ingestion to medical care was 12.4 hours (SD: 11.1). The mean initial ASA level was 48.3 mg/dl (SD: 16.4) with 86.9% having initial level >/=30mg/dl and 40.4% >/=50 mg/dl. 85.9% of cases presented to the hospital with their ASA level at or past peak. The mean pH was 7.45, pO2 = 108, pCO2 = 28.0, and HCO3 = 19.9. CONCLUSION: In this limited study, 85.9% of patients presenting with tinnitus and/or hearing loss following a single salicylate ingestion had initial salicylate levels at or past their peak and 86.9% were in the toxic range. PMID- 19561731 TI - Comparison of methanol exposure routes reported to Texas poison control centers. AB - OBJECTIVE: Methanol poisoning by ingestion is well represented in current emergency medicine literature. Much less described, however, is poisoning via intentional inhalation of methanol-containing products such as carburetor cleaner. This study intends to explore the exposure routes and treatment patterns of methanol cases reported to Texas Poison Centers. METHODS: All cases of methanol exposures from January 2003 to May 2005 were collected from the Texas Poison Center Network database "Toxicall." Inclusion criteria were 1) methanol as primary exposure, and 2) documented route of exposure. Exclusion criteria were unknown, dermal, and eye exposures. Data was extracted from documented calls to Texas Poison Centers and analyzed using descriptive statistics. RESULTS: A total of 203 cases were collected from 6 regional Poison Centers. Eighty seven cases had inhalation as the route of exposure, while 81 were methanol ingestions. Carburetor cleaner was responsible for nearly all the inhalational cases (79/87) while ingestions involved mostly windshield washer fluid (39/81) and carburetor cleaner (20/81). Seventy-eight percent of the inhalational exposures were intentional while most of the ingestions were accidental (49/75) and suicidal (18/75). An anion gap was documented in 31 of the inhalational cases and in 10 of the ingestions. Dialysis, use of fomepizole, and vision loss were documented for both types of exposure. Fifty-six percent of the inhalational group was admitted compared to 46% of the ingestion group. CONCLUSION: We propose that the results obtained from our review show inhalational exposure involving methanol (e.g., "huffing") represents a significant source of toxicity in the studied population. This is in contrast to previous literature that proposed inhalational toxicity was rare and aggressive treatment usually not necessary in cases of inhalation of methanol-containing carburetor cleaners. PMID- 19561732 TI - Fluoxetine overdose-induced seizure. AB - A 37-year-old woman experienced a witnessed generalized seizure in the Emergency Department three hours after ingesting approximately 1400 mg of fluoxetine in a suicide attempt. Although the majority of fluoxetine ingestions are benign, seizures may occur after large intentional overdoses. PMID- 19561733 TI - Survival after an intentional ingestion of crushed abrus seeds. AB - Abrus precatorius seeds contain one of the most potent toxins known to man. However, because of the seed's outer hard coat the vast majority of ingestions cause only mild symptoms and typically results in complete recovery. If the seeds are crushed and then ingested, more serious toxicity, including death, can occur.We present a case of a man who survived an intentional ingestion of crushed Abrus seeds after he was treated with aggressive gastric decontamination and supportive care. PMID- 19561734 TI - Fatal metformin overdose presenting with progressive hyperglycemia. AB - A 29-year-old man with no history of diabetes ingested over 60 grams of metformin in a suicide attempt. He presented to the emergency department with acute renal insufficiency, severe lactic acidosis, and rapidly-progressive hyperglycemia. The patient's peak serum glucose level of 707 mg/dL is the highest yet reported in a case of metformin toxicity. Treatment included sodium bicarbonate infusion and hemodialysis, but the patient suffered several cardiac arrests with pulseless electrical activity and ultimately expired 25 hours after the ingestion. PMID- 19561735 TI - Chemical pneumonitis from hydrocarbon aspiration. PMID- 19561736 TI - Issues and Solutions in Introducing Western Systems to the Pre-hospital Care System in Japan. AB - OBJECTIVE: This report aims to illustrate the history and current status of Japanese emergency medical services (EMS), including development of the specialty and characteristics adapted from the U.S. and European models. In addition, recommendations are made for improvement of the current systems. METHODS: Government reports and academic papers were reviewed, along with the collective experiences of the authors. Literature searches were performed in PubMed (English) and Ichushi (Japanese), using keywords such as emergency medicine and pre-hospital care. More recent and peer-reviewed articles were given priority in the selection process. RESULTS: The pre-hospital care system in Japan has developed as a mixture of U.S. and European systems. Other countries undergoing economic and industrial development similar to Japan may benefit from emulating the Japanese EMS model. DISCUSSION: Currently, the Japanese system is in transition, searching for the most suitable and efficient way of providing quality pre-hospital care. CONCLUSION: Japan has the potential to enhance its current pre-hospital care system, but this will require greater collaboration between physicians and paramedics, increased paramedic scope of medical practice, and greater Japanese societal recognition and support of paramedics. PMID- 19561737 TI - A Case of Complicated Urinary Tract Infection: Klebsiella pneumoniae Emphysematous Cystitis Presenting as Abdominal Pain in the Emergency Department. AB - THIS CASE REPORT DESCRIBES AN ATYPICAL PRESENTATION OF AN ATYPICAL DISEASE ENTITY: Emphysematous Cystitis, a rapidly progressive, ascending urinary tract infection, in an emergency department (ED) patient whose chief complaint was abdominal pain and who had a urinalysis not consistent with the diagnosis of cystitis. PMID- 19561738 TI - Herpes zoster ophthalmicus. PMID- 19561739 TI - Complications of MRSA Treatment: Linezolid-induced Myelosuppression Presenting with Pancytopenia. AB - Methicillin-resistant Staphylococcus aureus (MRSA) infections have grown to epidemic proportions in the United States. With the development of increasing drug resistance of MRSA to traditional antimicrobials, there has been a search for a more effective antibiotic treatment. Linezolid is one of the most effective oral medications used for outpatient treatment of MRSA infections. We present a case of pancytopenia after outpatient treatment with linezolid. Myelosuppression is a rare but serious side effect of linezolid of which emergency physicians need to be aware in order to provide early intervention. PMID- 19561740 TI - Images in emergency medicine: splenic infarction due to sickle cell trait after climbing mt. Fuji. PMID- 19561741 TI - Images in emergency medicine: pelvic digit. PMID- 19561742 TI - Emergent endotracheal intubation and mortality in traumatic brain injury. AB - OBJECTIVES: To determine the relationship between emergent intubation (emergency department and field intubation cases combined) and mortality in patients with traumatic brain injury while controlling for injury severity. METHODS: Retrospective observational study of 981 (35.2% intubated, 64.8% not intubated) patients with TBI evaluating the association between intubation status and mortality. Logistic regression was used to analyze the data. Injury severity measures included Head/Neck Abbreviated Injury Scale (H-AIS), systolic blood pressure, type of head injury (blunt vs. penetrating), and a propensity score combining the effects of several other potential confounding variables. Age was also included in the model. RESULTS: The simple association of emergent endotracheal intubation with death had an odds ratio (OR) of 14.3 (95% CI = 9.4 21.9). The logistic regression model including relevant covariates and a propensity score that adjusted for injury severity and age yielded an OR of 5.9 (95% CI = 3.2-10.9). CONCLUSIONS: This study indicates that emergent intubation is associated with increased risk of death after controlling for a number of injury severity indicators. We discuss the need for optimal paramedic training, and an understanding of the factors that guide patient selection and the decision to intubate in the field. PMID- 19561743 TI - Delayed complications of emergency airway management: a study of 533 emergency department intubations. AB - OBJECTIVES: Airway management is a critical procedure performed frequently in emergency departments (EDs). Previous studies have evaluated the complications associated with this procedure but have focused only on the immediate complications. The purpose of this study is to determine the incidence and nature of delayed complications of tracheal intubation performed in the ED at an academic center where intubations are performed by emergency physicians (EPs). METHODS: All tracheal intubations performed in the ED over a one-year period were identified; 540 tracheal intubations were performed during the study period. Of these, 523 charts (96.9%) were available for review and were retrospectively examined. Using a structured datasheet, delayed complications occurring within seven days of intubation were abstracted from the medical record. Charts were scrutinized for the following complications: acute myocardial infarction (MI), stroke, airway trauma from the intubation, and new respiratory infections. An additional 30 consecutive intubations were examined for the same complications in a prospective arm over a 29-day period. RESULTS: The overall success rate for tracheal intubation in the entire study group was 99.3% (549/553). Three patients who could not be orally intubated underwent emergent cricothyrotomy. Thus, the airway was successfully secured in 99.8% (552/553) of the patients requiring intubation. One patient, a seven-month-old infant, had unanticipated subglottic stenosis and could not be intubated by the emergency medicine attending or the anesthesiology attending. The patient was mask ventilated and was transported to the operating room for an emergent tracheotomy. Thirty-four patients (6.2% [95% CI 4.3 - 8.5%]) developed a new respiratory infection within seven days of intubation. Only 18 patients (3.3% [95% CI 1.9 - 5.1%]) had evidence of a new respiratory infection within 48 hours, indicating possible aspiration pneumonia secondary to airway management. Three patients (0.5% [95% CI 0.1 - 1.6%]) suffered an acute MI, but none appeared to be related to the intubation. One patient was having an acute MI at the time of intubation and the other two patients had MIs more than 24 hours after the intubation. No patient suffered a stroke (0% [95% CI 0 - 0.6%]). No patients suffered any serious airway trauma such as a laryngeal or vocal cord injury. CONCLUSIONS: Emergency tracheal intubation in the ED is associated with an extremely high success rate and a very low rate of delayed complications. Complication rates identified in this study compare favorably to reports of emergency intubations in other hospital settings. Tracheal intubation can safely be performed by trained EPs. PMID- 19561744 TI - The effect of single-bolus etomidate on septic patient mortality: a retrospective review. AB - BACKGROUND: Because of its many desirable properties, etomidate is widely used as an induction agent for endotracheal intubation. However, some have recently called into question the safety of etomidate for even single-bolus use due to its known effects on adrenal suppression. OBJECTIVES: We sought to compare the in hospital mortality between septic patients given etomidate and those given alternative induction agents for intubation. METHODS: We performed a retrospective chart review of intubated septic patients treated in our hospital. We collected data from patients over the age of 18 with sepsis who were intubated in the pre-hospital setting, in our emergency department, or on the wards of our hospital, and calculated the in-hospital mortality of each group. RESULTS: We identified 181 patients with sepsis who were intubated over the study period; 135 received etomidate and 46 received alternative agents or no induction agent. Baseline characteristics, vital signs, and laboratory values were similar between the two groups. Of the 46 patients receiving alternative agents or no agent, 18 died, yielding an unadjusted mortality of 39.1% (95% CI 25.5% to 54.6%), while of the 135 patients receiving etomidate, 63 died, for an unadjusted mortality of 46.7% (95% CI 38.1% to 55.4%), P=0.38. CONCLUSION: We found a non-statistically significant 7.6% absolute increase in mortality in patients given etomidate in our small-sized study population. PMID- 19561745 TI - Non-invasive method for the rapid assessment of central venous pressure: description and validation by a single examiner. AB - OBJECTIVES: This study describes a means of assessing the external jugular venous pressure (JVP) as an indicator of normal or elevated central venous pressure (CVP). METHODS: Intensive care unit patients having CVP monitoring were examined. With patients in bed, the external jugular vein (EJV) was occluded at the base of the neck and observed to distend. The occlusion was then removed and the vein observed for collapse. Complete collapse was hypothesized to indicate a non elevated CVP (8cm of water). RESULTS: In 12 of the 40 patients examined, the EJV could not be assessed (EJV not seen at all: 5, and difficult to visualize: 7). For the remaining 28 patients 11 had a CVP > 8 cm, while 17 had a CVP patients, of 2 mm and in locoregional metastasis. If distant metastasis is present and R0 surgery is not an option, the treatment should primarily comprise monochemotherapy or alternatively the patient should be enrolled in a clinical trial. CONCLUSION: The recommendations presented here are based predominantly on the results of prospective randomized trials. PMID- 19561814 TI - Hypermobility syndrome. PMID- 19561816 TI - Patients should be referred to centers. PMID- 19561812 TI - Tonsillectomy in children. AB - INTRODUCTION: Tonsillectomy is one of the most frequently performed surgical interventions in children. In the following, indications, preoperative evaluation, surgical techniques and postoperative complications will be discussed. METHODS: Literature search in PubMed (National Library of Medicine) focusing on publications in German or English up to June 2008. RESULTS: Indications are selected infectious diseases, upper airway obstruction for example due to tonsillar hypertrophy, and a suspected malignancy. Viral infections of the tonsils without upper airway obstruction are not an indication for surgery; in the case of acute bacterial tonsillitis, tonsillectomy is no longer recommended. In recurrent tonsillitis, tonsillectomy is only effective in specific and narrow indications. The indication for tonsillectomy in sleep disordered breathing due to adenotonsillar hypertrophy has to be based on clinical assessment, medical history, and a sleep history. The most relevant risk factors are obstructive sleep apnea and coagulation disorders. A standardized history regarding hemostasis and bleeding is mandatory, and is superior to routine coagulation tests. Postoperative bleeding is still the most relevant complication of tonsillectomy and is always an emergency situation. CONCLUSION: Tonsillectomy is one of the most frequently performed interventions in children but should be considered with care, as life-threatening complications can occur. PMID- 19561817 TI - Comprehensive literature. PMID- 19561819 TI - Painful red eye. PMID- 19561820 TI - Crash injury prediction and vehicle damage reporting by paramedics. AB - OBJECTIVE: The accuracy of pre-hospital crash scene details and crash victim assessment has important implications for initial trauma care assessment and management. Similarly, it is known to influence physician perception of crash victim injury severity. The goal of this feasibility study was to examine paramedic accuracy in predicting crash victim injury profile, disability outcome at hospital discharge, and reporting vehicle damage with other crash variables. METHODS: This prospective case series study was undertaken at a Southern California, Level I trauma center certified by the American College of Surgeons. Paramedics transporting crash injured motor vehicle occupants to our emergency department (ED)/trauma center were surveyed. We abstracted ED and in-patient records of injured vehicle occupants. Vehicle and crash scene data were obtained from a professional crash reconstruction, which included the assessment of deformation, crash forces, change in velocity, and the source of each injury. RESULTS: We used survey, injury, and crash reconstruction data from 22 collision cases in the final analysis. The median Injury Severity Score (ISS) was five (range 1-24). No enrolled patients died, and none were severely disabled at the time of discharge from the hospital. The paramedic crash injury severity predictions were sensitive for an Abbreviated Injury Scale (AIS) of 2-4. Paramedics often agreed with the crash reconstruction on restraint use, ejection, and other fatalities at the scene, and had lower levels of agreement for front airbag deployment, steering wheel damage, and window/windshield impact. Paramedics had 80% accuracy in predicting any disability at the time of hospital discharge. CONCLUSION: Paramedic prediction of injury profile was sensitive, and prediction of disability outcome at discharge was accurate when compared to discharge diagnosis. Their reporting of vehicle specific crash variables was less accurate. Further study should be undertaken to assess the benefits of crash biomechanics education for paramedics and other pre-hospital care providers. PMID- 19561821 TI - Unexpected arrest-related deaths in america: 12 months of open source surveillance. AB - INTRODUCTION: Sudden, unexpected arrest-related death (ARD) has been associated with drug abuse, extreme delirium or certain police practices. There is insufficient surveillance and causation data available. We report 12 months of surveillance data using a novel data collection methodology. METHODS: We used an open-source, prospective method to collect 12 consecutive months of data, including demographics, behavior, illicit substance use, control methods used, and time of collapse after law enforcement contact. Descriptive analysis and chi square testing were applied. RESULTS: There were 162 ARD events reported that met inclusion criteria. The majority were male with mean age 36 years, and involved bizarre, agitated behavior and reports of drug abuse just prior to death. Law enforcement control techniques included none (14%); empty-hand techniques (69%); intermediate weapons such as TASER((R)) device, impact weapon or chemical irritant spray (52%); and deadly force (12%). Time from contact to subject collapse included instantaneous (13%), within the first hour (53%) and 1-48 hours (35%). Significant collapse time associations occurred with the use of certain intermediate weapons. CONCLUSION: This surveillance report can be a foundation for discussing ARD. These data support the premise that ARDs primarily occur in persons with a certain demographic and behavior profile that includes middle-aged males exhibiting agitated, bizarre behavior generally following illicit drug abuse. Collapse time associations were demonstrated with the use of TASER devices and impact weapons. We recommend further study in this area to validate our data collection method and findings. PMID- 19561822 TI - Cervical spine motion during extrication: a pilot study. AB - Spinal immobilization is one of the most commonly performed pre-hospital procedures. Little research has been done on the movement of the neck during immobilization and extrication. In this study we used a sophisticated infrared six-camera motion-capture system (Motion Analysis Corporation, Santa Rosa, CA), to study the motion of the neck and head during extrication. A mock automobile was constructed to scale, and volunteer patients, with infrared markers on bony prominences, were extricated by experienced paramedics. We found in this pilot study that allowing an individual to exit the car under his own volition with cervical collar in place may result in the least amount of motion of the cervical spine. Further research should be conducted to verify these findings. In addition, this system could be utilized to study a variety of methods of extrication from automobile accidents. PMID- 19561823 TI - Factors associated with complications in older adults with isolated blunt chest trauma. AB - OBJECTIVE: To determine the prevalence of adverse events in elderly trauma patients with isolated blunt thoracic trauma, and to identify variables associated with these adverse events. METHODS: We performed a chart review of 160 trauma patients age 65 and older with significant blunt thoracic trauma, drawn from an American College of Surgeons Level I Trauma Center registry. Patients with serious injury to other body areas were excluded to prevent confounding the cause of adverse events. Adverse events were defined as acute respiratory distress syndrome or pneumonia, unanticipated intubation, transfer to the intensive care unit for hypoxemia, or death. Data collected included history, physical examination, radiographic findings, length of hospital stay, and clinical outcomes. RESULTS: Ninety-nine patients had isolated chest injury, while 61 others had other organ systems injured and were excluded. Sixteen patients developed adverse events [16.2% 95% confidence interval (CI) 9.5-24.9%], including two deaths. Adverse events were experienced by 19.2%, 6.1%, and 28.6% of those patients 65-74, 75-84, and >/=85 years old, respectively. The mean length of stay was 14.6 days in patients with an adverse event and 5.8 days in patients without. Post hoc analysis revealed that all 16 patients with an adverse event had one or more of the following: age >/=85, initial systolic blood pressure <90 mmHg, hemothorax, pneumothorax, three or more unilateral rib fractures, or pulmonary contusion (sensitivity 100%, CI 79.4-100%; specificity 38.6%, CI 28.1-49.9%). CONCLUSION: Adverse events from isolated thoracic trauma in elderly patients complicate 16% of our sample. These criteria were 100% sensitive and 38.5% specific for these adverse events. This study is a first step to identifying variables that might aid in identifying patients at high risk for serious adverse events. PMID- 19561825 TI - Pseudoaneurysm of the radial artery diagnosed by bedside ultrasound. AB - A 42-year-old male presented to the emergency department with pain and swelling of his distal right wrist. Bedside ultrasound placed over the swelling revealed a pseudoaneurysm of the radial artery. The patient received percutaneous thrombin injection of the aneurysm sac followed by direct ultrasound compression therapy of the pseudoaneurysm neck, resulting in thrombosis of the sac. The use of bedside ultrasound by the emergency physician led to appropriate care and proper disposition for definitive management. PMID- 19561824 TI - Analysis of urobilinogen and urine bilirubin for intra-abdominal injury in blunt trauma patients. AB - OBJECTIVE: To determine the point prevalence of urine bilirubin, urine hemoglobin and urobilinogen in blunt trauma patients, and to evaluate its utility as a screening tool for intra-abdominal injury. METHODS: Data analysis of 986 consecutive trauma patients of which 698 were adult blunt trauma patients. Five hundred sixteen subjects had a urinalysis and a CT scan of the abdomen/pelvis or exploratory laparotomy. We reviewed initial urinalysis results from trauma patients in the emergency department (ED) for the presence of urine hemoglobin, uroblinogen and urine bilirubin. Computed tomography (CT) scan results and operative reports were reviewed from the trauma registry for evidence of liver laceration, spleen laceration, bowel or mesenteric injuries. RESULTS: There were 73 injuries and 57/516 patients (11%) with intra-abdominal injury. Urinalysis was positive for urobilinogen in 28/516 (5.4%) patients, urine bilirubin in 15/516 (2.9%) patients and urine hemoglobin in 313/516 (61%) patients. Nineteen/forty seven (4%) subjects had liver lacerations, 28/56 (5%) splenic lacerations, and 15/5 (3%) bowel or mesenteric injury. Comparing the proportion of patients that had urobilinogen detected in the group with and without intra-abdominal injury, 8/28 (29%) subjects with urobilinogen, 5/15 (33%) subjects with bilirubin and 47/313 (15%) subjects with urine hemoglobin were found to have liver lacerations, spleen lacerations, or bowel/mesenteric injuries. Preexisting liver or biliary conditions were not statistically associated with elevation of urine bilirubin, urine hemoglobin or urobilinogen on initial urinalysis after blunt abdominal trauma. Point prevalence for urobilinogen, urine bilirubin and urine hemoglobin are 5.43% (28/516), 2.91% (15/516) and 60.7% (313/516) respectively. CONCLUSIONS: The utility of the initial routine urinalysis in the ED for adult blunt abdominal trauma patients should not be used as a screening tool for the evaluation of intra-abdominal injury. PMID- 19561826 TI - Subtalar dislocation. PMID- 19561827 TI - Higher inpatient medical surgical bed occupancy extends admitted patients' stay. AB - OBJECTIVE: Determine the effect that increased medical surgical (med/surg) bed occupancy has on the time interval from admission order to arrival in the bed for the patients admitted from the emergency department (ED). METHODS: This retrospective observational study compares the total hospital bed occupancy rate and the medical surgical inpatient bed occupancy rate to daily averages for the time interval from admission order (patient posting for admission) to the patient's arrival in the inpatient bed. Medical surgical inpatient bed occupancy of 92% was chosen because beyond that rate we observed more frequent extended daily transfer times. The data is from a single large tertiary care institute with 590 beds and an annual ED census of 80,000. RESULTS: Group 1 includes 38 days with (med/surg) inpatient bed occupancy rate of less than 92%, with an average ED daily wait of 2.5 hrs (95% confidence interval 2.23-2.96) for transfer from the ED to the appropriate hospital bed. Group 2 includes 68 days with med/surg census greater than 92% with an average ED daily wait of 4.1 hours (95% confidence interval 3.7-4.5). Minimum daily average for the two groups was 1.2 hrs and 1.3 hrs, respectively. The maximum average was 5.6 hrs for group 1 and 8.6 hrs for group 2. Comparison of group 1 to 2 for wait time to hospital bed yielded p <0.01. Total reported hospital occupied capacity shows a correlation coefficient of 0.16 to transfer time interval, which indicates a weak relationship between total occupancy and transfer time into the hospital. Med/surg occupancy, the beds typically used by ED patients, has a 0.62 correlation coefficient for a moderately strong relationship. CONCLUSIONS: Med/surg bed occupancy has a better correlation to extended transfer times, and occupancy over 92% at 5 AM in our institution corresponds to an increased frequency of extended transfer times from the ED. The process of ED evaluation, hospital admission, and subsequent transfer into the hospital are all complex processes. This study begins to demonstrate one variable, med/surg occupancy, as one of the intervals that can be followed to evaluate the process of ED admission and hospital flow. PMID- 19561828 TI - Routine laboratory testing to evaluate for medical illness in psychiatric patients in the emergency department is largely unrevealing. AB - OBJECTIVES: This is a prospective study of psychiatric patients presenting to the emergency department (ED) to determine the value of routine laboratory studies used to attempt to exclude concomitant medical illness. METHODS: Physical exams and laboratory tests were performed on 375 psychiatric patients presenting for "medical clearance" in the ED. Upon completion of these tests, the percentage and impact of abnormal physical exams and laboratory results were assessed. RESULTS: Fifty-six of 375 patients (14.9%) had a non-substance-induced laboratory abnormality. Forty-two of these 56 patients (75.0%) also had abnormal history or physical exam findings indicating laboratory screening. Ten had normal history and physical exams with insignificant laboratory abnormalities. The four (1.1% [95% CI 0.3-2.7%]) remaining patients with normal history and physical exams had abnormal urinalyses which did not affect final disposition or contribute to altered behavior. CONCLUSION: Patients presenting to the ED with psychiatric chief complaints, benign histories and normal physical exams have a low likelihood of clinically significant laboratory findings. PMID- 19561829 TI - Male patient visits to the emergency department decline during the play of major sporting events. AB - OBJECTIVES: To study whether emergency department (ED) visits by male patients wane simultaneously with the play of scheduled professional and college sports events. METHODS: Retrospective cohort analysis looked at ED male patient registration rates during a time block lasting from two hours before, during, and two hours after the play of professional football games (Monday night, Sundays, post-season play), major league baseball, and a Division I college football and basketball team, respectively. These registration rates were compared to rates at similar times on similar days of the week during the year devoid of a major sporting contest. Games were assumed to have a play time of three hours. Data was collected from April 2000 through March 2003 at an urban academic ED seeing 33,000 male patients above the age of 18 years annually. RESULTS: A total of 782 games were identified and used for purposes of the study. Professional football game dates had a mean of 17.9 males (95% confidence interval [CI] 17.4-18.4) registering vs. 26.8 males (95% CI 25.9-27.6) on non-game days. A registration rate for major league baseball was 18.4 patients (95% CI 17.6-18.4). The mean for registration on comparable non-game days was 23.9 patients (95% CI 22.8-24.3). For the regional Division I college football team, the mean number of patients registering on game days and non-game days was 21.7 (95% CI 20.9-22.4) and 23.4 (95% CI 22.9-23.7), respectively. Division I college basketball play for game and non-game days had mean rates of registration of 14.5 (95% CI 13.9-15.1) and 15.5 (95% CI 15.1-15.9) patients, respectively. For all sports dates collectively, a comparison of two means yielded a mean of 18.2 patients (95% CI 17.4-18.8) registering during the study hours on game days vs. 23.3 patients (95% CI 22.0 23.7) on non-game days. The mean difference was 5.1 patients (95% CI 3.7 to 7.0) with p < .000074. CONCLUSION: Male patient visits to the ED decline during major sporting events. PMID- 19561830 TI - Analysis of the literature on emergency department throughput. AB - INTRODUCTION: The purpose of this paper was to review and analyze all the literature concerning ED patient throughput. The secondary goal was to determine if certain factors would significantly alter patients' ED throughput. METHODS: A MEDLINE search was performed from 1966 to 2007 using the terms "turnaround," "emergency departments," "emergency medicine," "efficiency," "throughput," "overcrowding" and "crowding." Studies were graded using a scale of one to four based on the ACEP paper quality criteria. Inclusion criteria were English language and at least a level four or better on the quality scale. An analysis of successful procedures and techniques was performed. RESULTS: Literature search using the key terms found 29 articles on turnaround times, 129 on ED efficiency, 3 on throughput, 64 on overcrowding and 52 on crowding. Twenty-six articles were found to meet the inclusion criteria. There were three level I studies, thirteen level II studies, five level III studies and five level IV studies. The studies were categorized into five areas: determinants (7), laboratories processes (4), triage process (3), academic responsibilities (2), and techniques (10). Few papers used the same techniques or process to examine or reduce patient throughput precluding a meta-analysis. CONCLUSIONS: An analysis of the literature was difficult because of varying study methodologies and less than ideal quality. EDs with combinations of low inpatient census, in-room registration, point of care testing and an urgent care area demonstrated increased patient throughput. PMID- 19561831 TI - Supraclavicular subclavian vein catheterization: the forgotten central line. AB - While the supraclavicular approach to the subclavian vein has been described since 1965, it is generally employed much less often than the "traditional" infraclavicular approach. Although randomized trials are lacking, the best evidence suggests that the supraclavicular approach has a number of important advantages to the infraclavicular approach. The landmarks and relative merits of the procedure are described in this paper. PMID- 19561832 TI - Hemopericardium and cardiac tamponade in a patient with an elevated international normalized ratio. AB - This case report describes a 54-year-old male on warfarin for atrial fibrillation who presented to the emergency department (ED) following a syncopal episode with persistent hypotension. The patient's International Normalized Ratio (INR) returned elevated at 6.0, and a rapid bedside cardiac ultrasound revealed a large pericardial effusion consistent with cardiac tamponade. The anticoagulation was reversed and the patient underwent successful pericardiocentesis with removal of 1,100 mL of blood. PMID- 19561833 TI - Turning your abstract into a paper:academic writing made simpler. AB - Academic writing is a critical skill distinct from creative writing. While brevity is vital, clarity in writing reflects clarity of thought. This paper is a primer for novice academic writers. PMID- 19561835 TI - National priorities and goals. PMID- 19561834 TI - Challenging the cost effectiveness of medi-cal managed care. PMID- 19561836 TI - End the "ice age": is glacial acetic Acid really needed? PMID- 19561838 TI - New drugs drug news. PMID- 19561837 TI - Pharmacy distribution of consumer drug information emerges as a problem: FDA to seek solutions. PMID- 19561840 TI - Pushing an Expanded Role for Pharmacists: Medicare, SCHIP, and Health Insurance Debates Present Ample Opportunities. PMID- 19561839 TI - Bendamustine (treanda) for chronic lymphocytic leukemia: a brief overview. PMID- 19561841 TI - Better Asthma Management with Advanced Technology: Creation of an Asthma Utilization Rx Analyzer (AURA) Tool. AB - With nearly 23 million people affected by asthma each year, optimizing care among patients with persistent disease is a constant challenge for health care providers. The Asthma Utilization Rx Analyzer (AURA) tool enables health plan managers to evaluate quality and resource utilization for its members with asthma by analyzing medical and pharmacy claims. Customizable quality measures allow users of the tool to generate results from specific plans in order to optimize asthma disease management. PMID- 19561842 TI - Pharmaceutical approval update. PMID- 19561843 TI - 58th annual meeting american society of human genetics, 2008. AB - The 2008 ASHG meeting, which took place from November 11 to 15, brought almost 6,500 attendees to Philadelphia to learn about the latest advances in human genetics and genomics research. This conference is considered the world's largest gathering of human genetics researchers, academicians, clinicians, genetic counselors, and nurses. This article reviews the 1,000 Genomes Project, Gaucher's disease, warfarin dosing, and Huntington's disease. PMID- 19561844 TI - American society of hematology, 50th annual meeting and exposition. PMID- 19561845 TI - Evidence-informed case rates. PMID- 19561846 TI - An exhausted workforce increases the risk of errors. PMID- 19561847 TI - New FDA guidance on off-label promotion falls short for everyone: obama administration is likely to revisit it. PMID- 19561848 TI - New drugs drug news. PMID- 19561849 TI - Rilonacept (arcalyst), an interleukin-1 trap for the treatment of cryopyrin associated periodic syndromes. PMID- 19561850 TI - Administration of proton pump inhibitors in patients requiring enteral nutrition. PMID- 19561852 TI - FDA advisory committee meeting on prasugrel for acute coronary syndromes. PMID- 19561851 TI - Health care blog watch: emerging trends in the blogosphere. PMID- 19561853 TI - Pharmaceutical approval update. PMID- 19561854 TI - R&D redux. PMID- 19561855 TI - Preventing serious tissue injury with intravenous promethazine (phenergan). PMID- 19561856 TI - Comparative effectiveness research produces contentious debate-to what end? PMID- 19561857 TI - New drugs drug news. PMID- 19561858 TI - Nebivolol (bystolic), a novel Beta blocker for hypertension. PMID- 19561859 TI - Physician rights to privacy of data prevail in two major court tests, but new questions lie ahead. PMID- 19561860 TI - Pharmacoeconomic Analysis of Micafungin (Mycamine) 100 mg and 150 mg Daily In the Treatment of Candidemia. AB - We conducted a retrospective study to assess pharmacoeconomic outcomes of patients who received a daily dose of micafungin 100 mg or 150 mg to treat candidemia. The once-daily 100-mg dose resulted in clinical and mycological outcomes similar to those achieved with 150 mg daily and succeeded in reducing drug-acquisition costs for treating hospitalized patients with candidemia. PMID- 19561861 TI - Pharmaceutical approval update. PMID- 19561862 TI - Integrative healthcare symposium: cancer and chronic lyme disease. PMID- 19561863 TI - Spontaneous bacterial peritonitis: a review of treatment options. PMID- 19561864 TI - Double decade drivers. PMID- 19561865 TI - For best quality, send only original orders to pharmacies. PMID- 19561866 TI - Spotlight shines once again on medicaid payments to pharmacies: report and lawsuit settlement might lead to new formula. PMID- 19561867 TI - New drugs drug news. PMID- 19561868 TI - Rivaroxaban (xarelto) for the prevention of thromboembolic disease: an inside look at the oral direct factor xa inhibitor. PMID- 19561869 TI - When can patients sue drug companies?: supreme court finds that FDA approval does not stand in the way. PMID- 19561870 TI - Stimulus bill promises a new era in digital health care: but american recovery and reinvestment act gives pharmacies short shrift. PMID- 19561872 TI - American college of cardiology, 58th annual scientific session. PMID- 19561871 TI - H.p. Acthar gel and cosyntropin review: clinical and financial implications. PMID- 19561873 TI - Pharmaceutical approval update. PMID- 19561874 TI - Pulmonary hypertension: past, present and future. PMID- 19561875 TI - The presence of atypical mycobacteria in the mouthwashes of normal subjects: role of tap water and oral hygiene. AB - BACKGROUND: The nontuberculous mycobacteria (NTM) have been found in different environmental sources. They tend to colonize different body surfaces and secretions. The purpose of this study is to evaluate the presence of NTM in the oral cavity of healthy individuals and its relationship to tap water or oral hygiene. MATERIALS AND METHODS: One hundred sixty-seven healthy subjects were recruited. Three consecutive early morning mouthwashes using tap water were performed and examined for the presence of Mycobacterium tuberculosis (MTB) and NTM. In addition we obtained mouthwashes from 30 control healthy individuals with good oral hygiene using sterile water and examined these for the presence of MTB and NTM. RESULTS: NTM was isolated from the mouthwash of 44 (26.3%) subjects that used tap water. On the other hand, NTM was isolated from the mouthwash of 10 (33%) subjects that used sterile water. Age, gender, social class oral hygiene and the regular use of toothbrush made no statistically significant differences in the isolation rate of NTM. CONCLUSION: The rate of isolation of NTM from mouthwash is high in normal subjects. It is independent of oral hygiene, the use of tap water or teeth brushing. Smear-positive sputum could be NTM rather than M. tuberculosis. Tuberculosis polymerase chain reaction or culture confirmation is essential in developing countries to avoid the unnecessary use of antituberculosis therapy when the clinical suspicion is very low. PMID- 19561876 TI - Comparison of the efficacy and safety of video-assisted thoracoscopic surgery with the open method for the treatment of primary pneumothorax in adults. AB - OBJECTIVE: To determine whether video-assisted thoracoscopic surgery is as effective as the traditional open method through axillary thoracotomy for the management of primary spontaneous pneumothorax in adults. MATERIALS AND METHODS: This retrospective study was conducted at King Hussein Medical Center in the period between March 2002 and March 2007. Eighty-two patients were included in this study. The patients were divided in two groups: group A, which included patients who underwent video-assisted thoracoscopic surgery; and group B, which included patients who underwent open technique through axillary thoracotomy. Efficiency of the procedure, operative time, postoperative complications, length of hospital stay, time to return to work and recurrence were compared between the two groups. RESULTS: There were 79 males (96.3%) and 3 females (3.7%) with a mean age of 23.7 +/- 4.2 years for group A patients and 24.2 +/- 4.6 years for group B patients (range 16-37 years). Forty-one patients (50%) underwent video-assisted thoracoscopic surgery (group A), and 41 patients (50%) underwent open surgical technique through axillary thoracotomy (group B). Postoperative complication occurred in 8 patients (19.3%) from among those who underwent open technique and 6 patients (14.6%) from among those who underwent thoracoscopic technique. There was no perioperative mortality in both groups. Postoperative pain, volume of blood loss, period of air leak and hospital stay were less in group A, although the operative time was less in group B. CONCLUSION: Video-assisted thoracoscopic surgery is an efficient and safe method for the treatment of patients with primary pneumothorax in the adults. PMID- 19561877 TI - Percutaneous computed tomography-guided core biopsy for the diagnosis of mediastinal masses. AB - AIM: To describe various approaches of computed tomography (CT)-guided core biopsy and evaluate its ability to obtain adequate tissue for the assessment of mediastinal masses. MATERIALS AND METHODS: Between February 2004 and October 2006, 83 percutaneous CT-guided biopsies of mediastinal lesions were performed on 82 patients under local anesthesia. Coaxial needles were used and minimum of 3-4 cores were obtained. Post-biopsy CT scan was performed and patients observed for any complications. Tissue samples were taken to Pathology Department in formalin solution. RESULTS: From the 83 biopsies, adequate tissue for histological diagnosis was obtained in 80 (96%), and the biopsy was considered diagnostic. Of the 80 diagnostic biopsies, 74 biopsy samples were definitive for neoplastic pathology and 6 biopsy samples revealed no evidence of malignancy. There were no major complications. Minor complications were recorded in 5 patients. CONCLUSION: Percutaneous image-guided core biopsy of mediastinal lesions is an accurate, safe and cost-effective tool for the initial assessment of patients with mediastinal masses. PMID- 19561878 TI - Experience with 224 percutaneous dilatational tracheostomies at an adult intensive care unit in Bahrain: a descriptive study. AB - Tracheostomy is one of the most commonly performed procedures in critically ill patients. Over the past 15 years, many large university hospitals have reported their experience with percutaneous dilatational tracheostomy (PDT). We have described and compared our experience with 224 PDTs that we performed in the last four and a half years. We have also compared PDT performed with and without bronchoscopic guidance at our setting and PDT verses surgical tracheostomy. OBJECTIVE: The purposes of this study are to evaluate the safety of PDT in our hospital setting and to compare our results with those published in the literature. DESIGN: A retrospective study for our experiences about safety and efficacy of 224 PDTs in an intensive care unit (ICU) setting. SETTING: A 11 bedded adult medical, surgical, neuro-trauma ICU at Salmaniya Medical Complex, Bahrain. MATERIALS AND METHODS: This is based on our experiences about complications/timings of all PDTs performed from October 2002 to February 2007. A retrospective chart analysis. RESULTS: There were 15 mechanical complications in total, including nine patients developing bleeding during or post-procedure, three patients developing pneumothorax and two patients developing cellulitis; in one procedure, a tracheostomy tube was misplaced. The proportion of total complications was 6.6% and no death. CONCLUSIONS: From our experience, bronchoscope can be used during PDTs performed in ICU by inexperienced intensivists who do not have good exposure to procedures, but after gaining adequate experience, PDT can be performed safely without using bronchoscope. PMID- 19561879 TI - Acquired tracheoesophageal fistula due to high intracuff pressure. AB - High-compliance endotracheal tube cuffs are used to prevent gas leak and also pulmonary aspiration in mechanically ventilated patients. However, the use of the usual cuff inflation volumes may cause tracheal damage and lead to tracheoesophageal fistula.Tracheostomy tube cuffs seal against the tracheal wall and prevent leakage of air around the tube, assuring that the tidal volume is delivered to the lungs. In the past, high-pressure cuffs were used, but these contributed to tracheal injury and have been replaced by high-volume, low pressure cuffs. For long-term applications, some newer tubes have low-profile (tight to shaft) cuffs that facilitate the tracheostomy tube changes by eliminating the lip that forms when standard cuffs are deflated. PMID- 19561880 TI - Successful pregnancy outcome in Swyer-James-Macleod syndrome. AB - Swyer-James-MacLeod (SJM) syndrome is a chronic, progressive lung disease as a result of infection and bronchial obstruction that ultimately leads to emphysema. It is associated with chronic cough, sputum production and recurrent chest infections and is occasionally seen in women of reproductive age. The radiological finding of unilateral hyperlucent lung is considered synonymous with the disease entity. PMID- 19561881 TI - A young male with shortness of breath. AB - We report a case of primary mediastinal seminoma, which presented initially with shortness of breath and a swelling in upper part of anterior chest wall. The diagnosis of primary mediastinal seminoma was established on the basis of histologic findings and was confirmed by immunohistochemical analysis. Abdominal, pelvis and cerebral CT scan, testicular ultrasound and TC-99 MDP bone scintigraphy were negative. Chemotherapy was initiated with B.E.P. protocol (Bleomycin, Etoposide, Cisplatinum); the patient received four cycles of chemotherapy. After 8 months, the patient was seen in the clinic; he was well. PMID- 19561882 TI - Persistent cough in a 61-year-old male. PMID- 19561883 TI - Thoracic oncology multidisciplinary teams: between the promises and challenges. AB - The thoracic oncology multidisciplinary teams are playing an increasing role in the management of thoracic malignancies. These teams have a great potential to improve the patient care and the health care system, however, they are faced by many challenges. To realize the full potential of these teams, a better understanding of their functions, roles, benefits and challenges from all involved including teams members and leadership is crucial. PMID- 19561884 TI - International cooperation to promote advances in medicine. PMID- 19561885 TI - HRCT score in bronchiectasis: correlation with pulmonary function tests and pulmonary artery pressure. AB - BACKGROUND: High-resolution CT scan (HRCT) and its score have an important role in delineating pathological changes and pulmonary functional impairment in patients with bronchiectasis. AIMS: To assess pulmonary function tests (PFTs) in patients with cystic and cylindrical bronchiectasis. To correlate HRCT score with PFTs and systolic pulmonary artery pressure (SPAP) in both radiological types. MATERIALS AND METHODS: A cross-sectional study of patients with bronchiectasis diagnosed by HRCT was conducted at King Khalid University Hospital, Riyadh, Saudi Arabia. PFTs, HRCT score and SPAP were measured in both types. RESULTS: We studied 94 patients with bronchiectasis; 62 were cystic and 32 were cylindrical. Their mean age was 53.4 +/- 17.5 SD years. Forced vital capacity (FVC%) and forced expiratory volume in 1 second (FEV1%) were significantly lower in cystic patients (P < 0.0001) as compared with cylindrical patients; and diffusion capacity of carbon monoxide (DLCO%) was also significantly lower (P < 0.01). In the cystic group, PaO(2) was significantly lower; and PaCO(2), higher (P < 0.0001). HRCT score was correlated with FEV1% (r = -0.51). HRCT score was significantly lower in the cystic group (P = 0.002) and correlated with SPAP (r = 0.23). Global HTCT score of 10.3 +/- 2.5 was associated with SPAP > or = 40 mm Hg (P = 0.011). CONCLUSION: Patients with cystic bronchiectasis have significantly higher impairment of pulmonary physiology as compared with those with cylindrical bronchiectasis patients. HRCT score correlated with PFTs and SPAP. PMID- 19561886 TI - Correlation between high-resolution computed tomography features and patients' characteristics in chronic obstructive pulmonary disease. AB - BACKGROUND: During the last few decades, high-resolution computed tomography (HRCT) has come up as a new diagnostic modality to diagnose emphysematous and chronic bronchitis components of chronic obstructive pulmonary disease (COPD). The present study was undertaken to evaluate for various quantitative and qualitative HRCT features in patients with COPD, and to detect patients' characteristics that correlate with these HRCT features. MATERIALS AND METHODS: Forty male patients with COPD attending the COPD clinic at a tertiary referral hospital and postgraduate medical institute were included in the study. Various HRCT features, including tracheal index, thoracic cage ratio, sterno-aortic distance, thoracic cross-sectional area, vascular attenuation, vascular distortion, mosaic attenuation pattern, and directly visible small airways, were evaluated and correlated with patients' characteristics, including age, duration of illness, quantum of smoking, dyspnea score, quality-of-life index, and various spirometric indices. RESULTS: We found significant correlations of various quantitative and qualitative HRCT features with age, duration of illness, quantum of smoking, quality-of-life index, and the spirometric indices showing the extent of airways obstruction. CONCLUSIONS: Various quantitative and qualitative HRCT features were found to correlate with patients' characteristics, spirometric indices, and health-related quality-of-life score, suggesting that HRCT is useful not only in detecting emphysema and its various subtypes but also in predicting the extent and severity of COPD. PMID- 19561887 TI - Using arm span to derive height: impact of three estimates of height on interpretation of spirometry. AB - BACKGROUND: When standing height required to calculate forced vital capacity (FVC) cannot be measured, it can be derived from arm span using different methods. OBJECTIVES: To compare three different estimates of height derived from arm span and investigate their impact on interpretation of spirometric data. METHODS: In a cross-sectional study, 517 subjects aged 7 to 76 years, with various respiratory diseases underwent spirometry. Three estimates of height were obtained from arm span: (a) by direct substitution (Ht(AS)); (b) estimated height (Ht(est)), obtained from the mean arm span:standing height ratio; and (c) predicted height (Ht(pred)), obtained from arm span by linear regression analysis. Predicted values of forced vital capacity (FVC) obtained from these estimates were compared with those obtained from actual standing height (Ht(act)), followed by Bland Altman analysis of agreement in the patterns of ventilatory impairment. RESULTS: The arm span was 5%-6% greater than the height. The difference increased with increasing height. Ht(AS) and the FVC predicted from it were significantly greater than the other measures of height and the related predicted FVCs respectively. Compared to Ht(act), Ht(AS) gave a misclassification rate of 23.7% in taller subjects (Ht(act) > 150 cm) and 14.2% in shorter subjects in the patterns of ventilatory impairment. Misclassification rates were 6%-8% with Ht(est) and Ht(pred). Agreement analysis showed that FVCs predicted from Ht(pred) had the best agreement with the FVC predicted from Ht(act). CONCLUSIONS: Among several methods of estimating height from the arm span, prediction by regression is most appropriate as it gives least errors in interpretation of spirometric data. PMID- 19561888 TI - Role of nitric oxide in management of acute respiratory distress syndrome. AB - The current mortality rate of patients suffering from acute respiratory distress syndrome (ARDS) is between 45% and 92%, with most dying within the first two weeks of the illness. In an effort to combat such an alarmingly high mortality rate, various treatment therapies such as low tidal volume ventilation strategies, corticosteroid therapy, and use of nitric oxide (NO) have been attempted in the management of patients with ARDS. Three cases which were admitted to the ICU and confirmed to have ARDS were unable to be weaned from ventilatory support, and nitric oxide therapy was initiated. It improved patients' oxygenation for short periods of time but did not affect the mortality. The patients could not be weaned from the ventilator and expired. PMID- 19561889 TI - Spontaneous pneumothorax: an unusual complication of pregnancy--a case report and review of literature. AB - Spontaneous pneumothorax complicating pregnancy is rare. Only 55 cases have been reported till now. We describe a case of a 30-year-old Indian woman with spontaneous pneumothorax during her 28(th) week of pregnancy. PMID- 19561890 TI - Unusual variant of Cantrell's pentalogy? AB - A 12-hour-old male infant presented with prolapsed abdominal content through a defect on left side of chest wall with respiratory distress. A thorough clinical examination suggested absence of ectopia cordis, abdominal wall defect, and any bony anomaly. The child expired after 6 hours of admission because of respiratory distress and electrolyte imbalance. Is congenital defect of chest wall associated with diaphragmatic hernia without ectopia cordis and omphalocele, an unusual variant of Cantrell's pentalogy? PMID- 19561891 TI - Colopleural fistula: case report and review of the literature. AB - We report a 28-year-old woman, pregnant, at 24 weeks, with 3-day history of right sided chest pain and shortness of breath. Few hours after admission, she delivered a dead baby. She had a history of right partial hepatic lobotomy and cholecystectomy at UK on May 2004 because of multiple pyogenic liver abscesses. Chest examination revealed signs of hydrothorax on the right side. Chest X-ray showed pleural effusion on the right side. Pleural fluid was exudative with high neutrophils. Gram stain and culture showed multiple organisms. CT scan chest and abdomen with contrast, combined with barium enema, revealed right colothorax communication. Colothorax fistula was closed surgically. On the following days, the patient's symptoms resolved, and she was consequently discharged. PMID- 19561893 TI - The first Saudi lung cancer guidelines. PMID- 19561892 TI - Tuberculosis of the breast. AB - Tuberculosis of the breast is an uncommon disease even in countries where the incidence of pulmonary and extrapulmonary tuberculosis is high. Clinical presentation is usually of a solitary, ill-defined, unilateral hard lump situated in the upper outer quadrant of the breast. This disease can present a diagnostic problem on radiological and microbiological investigations, and thus a high index of suspicion is needed. Incorporating a highly sensitive technique like polymerase chain reaction (PCR) may be helpful in establishing the usefulness of such technology and can aid in conforming the diagnosis early. The disease is curable with antitubercular drugs, and surgery is rarely required. PMID- 19561894 TI - Evaluation of brain stem auditory evoked potentials in stable patients with chronic obstructive pulmonary disease. AB - Though there are few studies addressing brainstem auditory evoked potentials (BAEP) in patients with chronic obstructive pulmonary disease (COPD), subclinical BAEP abnormalities in stable COPD patients have not been studied. The present study aimed to evaluate the BAEP abnormalities in this study group. MATERIALS AND METHODS: In the present study, 80 male subjects were included: COPD group comprised 40 smokers with stable COPD with no clinical neuropathy; 40 age-matched healthy volunteers served as the control group. Latencies of BAEP waves I, II, III, IV, and V, together with interpeak latencies (IPLs) of I-III, I-V, and III V, and amplitudes of waves I-Ia and V-Va were studied in both the groups to compare the BAEP abnormalities in COPD group; the latter were correlated with patient characteristics and Mini-Mental Status Examination Questionnaire (MMSEQ) scores to seek any significant correlation. RESULTS: Twenty-six (65%) of the 40 COPD patients had BAEP abnormalities. We observed significantly prolonged latencies of waves I, III, V over left ear and waves III, IV, V over right ear; increased IPLs of I-V, III-V over left ear and of I-III, I-V, III-V over right side. Amplitudes of waves I-Ia and V-Va were decreased bilaterally. Over left ear, the latencies of wave I and III were significantly correlated with FEV(1); and amplitude of wave I-Ia, with smoking pack years. A weak positive correlation between amplitude of wave I-Ia and duration of illness; and a weak negative correlation between amplitude of wave V-Va and MMSEQ scores were seen over right side. CONCLUSIONS: We observed significant subclinical BAEP abnormalities on electrophysiological evaluation in studied stable COPD male patients having mild to-moderate airflow obstruction. PMID- 19561895 TI - Congenital bronchopulmonary malformations: a single-center experience and a review of literature. AB - PURPOSE: To present a single-center experience with 25 cases of bronchopulmonary malformations and the review the literature. MATERIALS AND METHODS: We conducted a retrospective analysis of the medical records of patients with congenital bronchopulmonary malformations who were operated between July 1997 and July 2007 in our institute; we examined the modes of presentations, management, and outcome. Outcome of all patients was assessed over a short follow-up period (average 1.8 months). RESULTS: Out of 25 patients, 18 (72%) were male and 7 (28%) were female. Age of patients ranged from 1 day to 11 years. The histopathological diagnosis was congenital cystic adenomatoid malformations [CCAM; n = 14 (56%)], congenital lobar emphysema [CLE; n = 5 (20%)], pulmonary sequestrations [PS; n = 3 (12%)], and bronchogenic cysts [BC; n = 3 (12%)]. Antenatal diagnosis was available in only 2 (8%) patients. The common presenting symptoms were respiratory distress and chest infections. Lobectomy was the procedure of choice. Mortality was 16% (n = 4; M: F = 3: 1). Two patients died because of overwhelming sepsis, one from compromised cardiac function, and one from aspiration which might possibly have been prevented. CONCLUSION: Patients with progressive respiratory distress due to these anomalies may require urgent surgical intervention regardless of age. The surgical outcome is favorable, with manageable complications. Plain x-ray chest and CT of thorax are usually sufficient for diagnosis and planning of treatment. Pathological diagnosis may differ from the imaging diagnosis. Mortality is found to be more in neonates. Apart from initial stabilization, resection of lesion and careful postoperative care is necessary to reduce mortality and morbidity. PMID- 19561896 TI - Comparison of clinicoradiologic manifestation of nonspecific interstitial pneumonia and usual interstitial pneumonia/idiopathic pulmonary fibrosis: a report from NRITLD. AB - BACKGROUND: Ever since Katzenstein and Fiorelli introduced the term nonspecific interstitial pneumonia (NSIP) to denote those cases of interstitial pneumonia that cannot be categorized as any of the other types of idiopathic interstitial pneumonias (IIP), there has been continuing debate on whether it is a real clinical entity or not. The American Thoracic Society/European Respiratory Symposium task group tried to identify idiopathic NSIP as a separate disease and exclude it from the category of IIP. However, it appears that the clinical presentation of NSIP and usual interstitial pneumonia (UIP) are the same. OBJECTIVE: To show that the radiologic features of NSIP and UIP should be relied upon, instead of clinical presentation and pathologic findings, to differentiate between the two. MATERIALS AND METHODS: Consecutive patients who had received a diagnosis of either NSIP or UIP on the basis of open lung biopsy between January 2001 and December 2007 were identified for inclusion in this retrospective review. The study included 61 subjects: 32 men and 29 women with a mean age of 59.39 +/- 14.5 years. Chest computed tomography images of all the cases were collected for a review. High resolution computed tomography (HRCT) and all pathologic specimens were also evaluated. A weighted kappa coefficient was used to evaluate whether radiology can be used instead of biopsy for the diagnosis of NSIP and UIP. Comparison of the mean ages and the time intervals (i.e., interval between symptom onset and the time of diagnosis) in the UIP and NSIP groups was done using the Mann-Whitney U test. Association between gender and biopsy result was evaluated by the Fisher exact test. Data were evaluated using SPSS, v.13. RESULTS: Sixty-one patients were included in this study, 32 were male and 29 were female. On the basis of biopsy findings, 50 (82%) patients had UIP and 11 (18%) had NSIP. Thirty (60%) of the 50 patients who had UIP were male and 20 (40%) were female; 2 (18.2%) of the 11 patients who suffered from NSIP were male and 9 (81.8%) were female. Based on HRCT findings, 36 (60%) patients were diagnosed to have UIP and 24 (40%) were diagnosed with NSIP. When diagnosis was based on biopsy findings, the time interval in the UIP group was 13.59 +/- 8.29 months and in the NSIP group it was 7.90 +/- 4.18 months. When diagnosed on the basis of HRCT findings, the time interval in the UIP group was 14.22 +/- 8.94 months and in the NSIP group it was 10.54 +/- 5.78 months. When diagnosis was on the basis of biopsy, the mean age in the UIP group was 61.30 +/- 14.18 years and in the NSIP group it was 50.73 +/- 13.14 years. CONCLUSION: HRCT can be used instead of invasive methods like lung biopsy to differentiate between UIP and NSIP. PMID- 19561898 TI - Isoniazid- and ethambutol-induced psychosis. AB - Most cases of antituberculous agent-associated psychoses were caused by isoniazid (INH), with ethambutol (EMB)-induced psychosis being rare. The concomitant occurrence of INH- and EMB- induced psychosis and in a single individual is extremely uncommon. We report a case of 28-year-old male who developed psychotic symptoms on start of EMB initially and later on INH also. He was prescribed rifampicin, pyrazinamide, and ofloxacin and had no further psychotic symptoms. PMID- 19561897 TI - Diagnosis of peripheral pulmonary carcinoid tumor using endobronchial ultrasound. AB - A 51-year-old woman with severe asthma underwent bronchoscopy and endobronchial ultrasound (EBUS) for investigation of a 15-mm peripheral lung nodule. Histology demonstrated a typical carcinoid tumor. Pulmonary location is the second commonest site for carcinoid tumors. Diagnosis of peripheral carcinoid tumor of the lung is difficult due to its small size, poor accuracy of cytologic diagnosis, and low sensitivity of positron emission tomography in detecting it. EBUS has a high diagnostic yield and a low complication rate in the evaluation of small solitary pulmonary nodules. The ultrasound appearance of carcinoid tumors is identical to that of lung carcinomas. Prompt diagnosis of carcinoid tumor is desirable as regional lymph node metastasis is seen in 10% of patients and is associated with a reduced 5-year survival. We feel that, where possible, all patients presenting with solitary pulmonary nodules should be investigated initially using EBUS due to its high diagnostic rate and the very low incidence of adverse events. PMID- 19561899 TI - Ectopia cordis associated with Cantrell's pentalogy. AB - Cantrell's pentalogy with ectopia cordis is an extremely rare and lethal congenital anomaly, with a reported incidence of 1:100,000 births in developed countries. We report a neonate who presented with ectopia cordis along with cleft lower sternum, upper abdominal wall defect, ectopic umbilicus, diaphragmatic defect, and interventricular septal defect. The neonate had respiratory distress with peripheral cyanosis and died because of acidosis and electrolyte imbalance before surgical intervention could be undertaken. We discuss the case and present a brief review of literature and of embryogenesis. PMID- 19561900 TI - Avian influenza: the tip of the iceberg. AB - For some years now, we have been living with the fear of an impending pandemic of avian influenza (AI). Despite the recognition, in 1996, of the global threat posed by the highly pathogenic H5N1 influenza virus found in farmed geese in Guangdong Province, China, planning for the anticipated epidemic remains woefully inadequate; this is especially true in developing countries such as Saudi Arabia. These deficiencies became obvious in 1997, with the outbreak of AI in the live animal markets in Hong Kong that led to the transmission of infection to 18 humans with close contact with diseased birds; there were six reported deaths. In 2003, with the reemergence of H5N1 (considered the most likely AI virus) in the Republic of Korea and its subsequent spread to Thailand, Vietnam, Hong Kong and China. Many countries started aggressively making preparations to meet the threat. The pressure for real action from governments has increased. Most developed countries have requested increased funding for the search for a more effective vaccine, for stockpiling possibly helpful antiviral drugs, and for intensifying domestic and global surveillance. Most countries, however, continue to be inadequately prepared for such an epidemic, especially with regard to animal surveillance in the farm market and surveillance among migratory birds. Even now, most countries do not have the ability to detect disease among humans in the early stages of an outbreak nor do most hospitals comply with effective infection control measures that could curtail the spread of the virus in the early stages of an epidemic. In Saudi Arabia we are rapidly implementing many of these measures. PMID- 19561901 TI - A female with fever and shortness of breath. PMID- 19561902 TI - Bronchial asthma in developing countries: a major social and economic burden. PMID- 19561903 TI - Does consanguinity increase the risk of bronchial asthma in children? AB - There is a high prevalence of consanguinity and bronchial asthma in Saudi Arabia. The objective of this study is to explore the effect of parental consanguinity on the occurrence of bronchial asthma in children. The study sample was determined by multistage random probability sampling of Saudi households. The families with at least one child with asthma were matched with an equal number of families randomly selected from a list of families with healthy children, the latter families being designated as controls. There were 103 families with children having physician-diagnosed bronchial asthma, matched with an equal number of families with no children with asthma. This resulted in 140 children with bronchial asthma and 295 children from controls. The age and gender distribution of the children with bronchial asthma and children from controls were similar. There were 54/103 (52.4%) and 61/103 (59.2%) cases of positive parental consanguinity in asthmatic children and children from controls respectively (P = 0.40). Analysis of consanguinity status of the parents of children with asthma and parents among controls indicates that 71/140 (51%) of the children with asthma and 163/295 (55.3%) of the children from controls had positive parental overall consanguinity (P = 0.43). The results of this study suggest that parental consanguinity does not increase the risk of bronchial asthma in children. PMID- 19561904 TI - Validation of the Arabic version of the asthma control test. AB - PURPOSE: Asthma control test (ACT) has been devised to assess the degree of asthma control in out-patients setting. The aim of this study is to validate the Arabic version of ACT. MATERIALS AND METHODS: Patients completed the Arabic version of ACT during regular visit to one of two asthma specialists. Spirometry was obtained. The asthma specialist rated asthma control using a 5-point scale and indicated modification in management as step up, same or step down of asthma treatment. RESULTS: 40 patients completed the study, the mean age was 32.6 + 14.0 years, mean FEV1 was 2.7 + 1.0 L (89.2% + 23.6% of predicted). The mean ACT score was 15.9 + 5.8; mean of specialist asthma control rating was 3.4 + 1.0. The internal consistency reliability of the 5-item ACT survey was alpha = 0.92. The correlation was moderate between ACT and specialists rating (r = 0.482, P = 0.002) and between ACT and treatment modification (r = -0.350, P = 0.027). The correlation between FEV1 and ACT was low (r = 0.185, P = 0.259). ACT distinguished between patients with different specialist rating (F = 3.37, P = 0.02) and the need to change therapy (F = 3.62, P = 0.037). The areas under the curve (ROC) for ACT, FEV1, and ACT and FEV1 as independent variables were 0.720, 0.721, and 0.766 respectively. All results were comparable to the initial work for development of ACT. CONCLUSION: The Arabic version of the ACT is a valid tool to assess asthma control. ACT correlates better with asthma specialist rating of asthma control than with FEV1. PMID- 19561905 TI - Unconventional therapy use among asthma patients in a tertiary care center in Riyadh, Saudi Arabia. AB - OBJECTIVES: Unconventional therapy (UT) is a therapeutic practice of alternative and complementary medicine that is not currently considered an integral part of modern medical practice. The aim of this article is to investigate the experience of Saudi patients with UT modalities in the treatment of asthma. MATERIALS AND METHODS: We carried out a cross-sectional study of asthma patients referred to King Abdulaziz Medical City, Riyadh, Saudi Arabia, during the year 2004. Information was collected using a pre-designed questionnaire administered through interviews. RESULTS: Two hundred consecutive patients with a mean age of 52.3 years (+/-18.7) were included in this study. Sixty-nine (34.5%) of those patients used some form of UT in the previous year. There was a tendency to use UT among the older age group (P = 0.029) and among those with longer duration of disease (P = 0.009). However, there was no significant correlation observed between the use of UT and gender, FEV(1), or disease control. The most commonly used form of UT was recitation of Holy Quran (9%), honey (24.5%), herbs (23.5%), cautery (12%), and blackseed (10%). There was no significant correlation between disease control and the use of modalities. CONCLUSION: Unconventional therapy is frequently practiced by asthma patients in Saudi Arabia, who commonly believe that UT will lead to improvement. The lack of evidence necessitates the fostering of a national project to address the practice of UT. PMID- 19561906 TI - Knowledge and practice of spirometry among pediatricians in Riyadh, Kingdom of Saudi Arabia. AB - BACKGROUND: Spirometry is the most basic, widely used and effort-dependent pulmonary function test. It assesses the lung volumes and flows, and is ideally suited to describe the effects of restriction or obstruction on lung function. Therefore, keeping in view the clinical applications of spirometry, this study attempts to explore the knowledge and practice about spirometry among pediatricians. MATERIALS AND METHODS: A questionnaire-based study was conducted across multiple centers in various hospitals in Riyadh, Saudi Arabia. The structured questionnaire, based upon knowledge and practice of spirometry, was distributed to 150 pediatricians in the various tertiary care hospitals in the metropolitan area of Riyadh. RESULTS: Ninety-four percent of 113 pediatricians agreed that spirometry is a valuable tool in pediatric clinical practice. However, knowledge relating to spirometry was lacking among pediatricians, and about 86% of the study population did not demonstrate up-to-date knowledge of spirometry in pediatrics. Only 11% of pediatricians were very confident in interpreting spirometry results. No statistically significant association was observed between the distribution of responses relating to knowledge and practice of spirometry and the study variables including academic position, duration of practicing experience and number of patients attended daily. CONCLUSION: The results indicated that pediatricians in Riyadh were lacking adequate knowledge about the clinical applications of spirometry in their daily clinical practice. Hence, it was suggested that pediatricians should attend periodical training, workshops and continuous medical education programmes to enhance their knowledge. This should especially be performed during their pediatric residency training programmes, as spirometry is one of the essential components in clinical practice. PMID- 19561907 TI - Hypocalcemia in a Saudi intensive care unit. AB - OBJECTIVE: Hypocalcemia has been a common abnormality in the West, seen in patients admitted to the intensive care unit (ICU). It has also been linked with disease severity. We undertook this study to determine the frequency of hypocalcemia in patients admitted to the intensive care unit. MATERIALS AND METHODS: In a retrospective chart review from January 2004 till December 2004, patients admitted to our ICU were reviewed. Patients' age, sex, diagnosis, acute physiology and chronic health score APACHE II and ionized calcium were recorded. Patients were divided into three groups based on disease severity as measured by APACHE II. Hypocalcemia was defined as ionized Ca level less than 1.18 mmol/L. Frequency was determined in each group and correlation of hypocalcemia with disease severity was explored. RESULTS: Hypocalcemia was seen in 22.2% in group A (APACHE II < 10), 40.4% in group B (APACHE II 10-19) and 53.9% in group C (APACHE II > 19). Hypocalcemia and disease severity (APACHE II scores) were negatively correlated (P = 0.02). Mean ionized Ca levels in groups A, B and C were 1.22 mmol/L (+/-0.10), 1.19 mmol/L (+/-0.11) and 1.25 mmol/L (+/-0.24) respectively. CONCLUSIONS: Hypocalcemia is a common finding in critically ill patients. It is correlated with worsening disease severity. Mechanisms underlying hypocalcemia and the possible relationship of hypocalcemia with mortality need further consideration. PMID- 19561908 TI - Hypogenetic lung syndrome in an adolescent: imaging findings with short review. AB - Hypogenetic lung syndrome is more popularly known as a scimitar syndrome (SS). It is a rare developmental lung malformation which almost always occurs on the right side. The two most constant features of this syndrome are anomalous pulmonary venous return into systemic circulation, most frequently via inferior vena cava (IVC), and lung hypoplasia.We are reporting such a case illustrating most typical and some uncommon features on chest radiograph and multislice computer tomography (MSCT) of chest. Focal herniation of liver through a diaphragmatic defect presenting as an ovoid soft tissue mass in right lower paraspinal region on chest X ray mimicking sequestration is an interesting but rare finding. PMID- 19561909 TI - Extramedullary paraspinal hematopoiesis in hereditary spherocytosis. AB - Hereditary spherocytosis (HS) is a common inherited hemolytic anemia due to red cell membrane defects. Extramedullary hematopoiesis is a compensatory response to insufficient bone marrow blood cell production. The preferred sites of extramedullary hematopoietic involvement are the spleen, liver and lymph nodes; but in HS, the posterior paravertebral mediastinum is also commonly involved. We report a case of a 50-year-old male who presented to us in respiratory distress and with bilateral paravertebral posterior mediastinal masses, which on trucut biopsy were found to be extra-hematopoietic masses; and the patient was found to have hereditary spherocytosis. PMID- 19561912 TI - Annals of Thoracic Medicine ... a three-year journey. PMID- 19561913 TI - Fayez never saw sunlight: There is a need for more specialized respiratory units. PMID- 19561910 TI - Bronchiolitis obliterans organizing pneumonia: pathogenesis, clinical features, imaging and therapy review. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) was first described in the early 1980s as a clinicopathologic syndrome characterized symptomatically by subacute or chronic respiratory illness and histopathologically by the presence of granulation tissue in the bronchiolar lumen, alveolar ducts and some alveoli, associated with a variable degree of interstitial and airspace infiltration by mononuclear cells and foamy macrophages. Persons of all ages can be affected. Dry cough and shortness of breath of 2 weeks to 2 months in duration usually characterizes BOOP. Symptoms persist despite antibiotic therapy. On imaging, air space consolidation can be indistinguishable from chronic eosinophilic pneumonia (CEP), interstitial pneumonitis (acute, nonspecific and usual interstitial pneumonitis, neoplasm, inflammation and infection). The definitive diagnosis is achieved by tissue biopsy. Patients with BOOP respond favorably to treatment with steroids. PMID- 19561914 TI - Diagnosis of latent tuberculosis: Can we do better? AB - Latent tuberculosis infection (LTBI) is often diagnosed by the tuberculin skin test (TST). The latter has several limitations with regard to its sensitivity and specificity. It may be positive in people with prior bacille Calmette-Guerin (BCG) vaccination or exposure to nontuberculous mycobacteria. False negative TST results frequently occur in patients with impaired T-cell function. Therefore TST results have to be interpreted taking into consideration the pretest risk of TB infection or reactivation. Recently, interferon gamma release assays (IGRA) were introduced for the diagnosis of LTBI. These include the T-SPOT-TB and the QuantiFERON((R))-TB Gold tests.These tests measure interferon gamma released in response to T-cell stimulation by specific Mycobacterium tuberculosis antigens. These tests have been shown to be more specific than the TST as they are not affected by BCG vaccination. Their sensitivity was similar to that of the TST and in some studies they correlated better with the degree of exposure. In immune compromised patients their sensitivity was better than that of the TST. IGRA tests were shown to have better predictive value for the development of active disease among individuals with LTBI. These tests are expensive. Their most cost effective utilization is as confirmatory tests in patients with positive TST results, particularly in areas with high rates of BCG vaccination. PMID- 19561915 TI - Lessons from patients with hemoptysis attending a chest clinic in India. AB - OBJECTIVE: To evaluate the various etiologies of hemoptysis. MATERIALS AND METHODS: Four hundred and seventy-six consecutive patients of hemoptysis who were admitted to the Department of Pulmonary Medicine between January 1996 and December 2002 were included in this study. Hemoptysis was categorized as mild (< 100 ml/day), moderate (100-400 ml/day), and massive (>400 ml/day). We also categorized the patients according to the primary etiology of the hemoptysis. RESULTS: Of the 476 patients with hemoptysis included in this study, 352 were males and 124 were females. Pulmonary tuberculosis was the leading cause of hemoptysis. There were 377 (79.2%) patients in the pulmonary tuberculosis group, 25 (5.7%) in the neoplasm group, 19 (4.0%) in the chronic bronchitis group, 18 (3.8%) in the bronchiectasis group, and 35 (7.3%) patients with hemoptysis due to other causes. About one-third of the patients with hemoptysis had been misdiagnosed by the referring doctor as having active pulmonary tuberculosis. CONCLUSION: Although pulmonary tuberculosis is the most important cause of hemoptysis in India, it may also occur due to a variety of other causes. Awareness should be increased among general physicians about the various etiologies of hemoptysis in pulmonary tuberculosis patients. PMID- 19561916 TI - Outcome of patients with pulmonary embolism admitted to the intensive care unit. AB - BACKGROUND: Pulmonary embolism (PE) is an important cause of in-hospital mortality. Many patients are admitted to the intensive care unit (ICU) either due to hemodynamic instability or severe hypoxemia. Few reports have addressed the outcome of patients with PE; however, none were from ICUs in the Middle East. OBJECTIVES: To describe the demographics, clinical presentation, risk factors and outcome of patients with PE admitted to the medical ICU and to identify possible factors associated with poor prognosis. MATERIALS AND METHODS: Data were collected retrospectively by reviewing the records of patients admitted to the medical ICU with primary diagnosis of PE between January 2001 and June 2007. Demographic, clinical, radiological and therapeutic data were collected on admission to ICU. RESULTS: Fifty-six patients (43% females) with PE were admitted to the ICU during the study period. Their mean age was 40.6 +/- 10.6 years. Seven patients (12.5%) had massive PE with hemodynamic instability and 15 (26.8%) had submassive PE. The remaining patients were admitted due to severe hypoxemia. Recent surgery followed by obesity were the most common risk factors (55.4 and 28.6%, respectively). Four patients with massive PE received thrombolysis because the remaining three had absolute contraindications. Fatal gastrointestinal bleeding occurred in one patient post thrombolysis. Additionally, two patients with massive PE and five with submassive PE died within 72 h of admission to the ICU, resulting in an overall mortality rate of 14%. Nonsurvivors were older and had a higher prevalence of immobility and cerebrovascular diseases compared with survivors. CONCLUSIONS: The mortality rate of patients with PE admitted to the ICU in our center was comparable to other published studies. Older age, immobility as well as coexistent cerebrovascular diseases were associated with a worse outcome. PMID- 19561917 TI - Noninvasive positive pressure ventilation in unplanned extubation. AB - BACKGROUND: Unplanned extubation is quite common in intensive care unit (ICU) patients receiving mechanical ventilatory support. The present study aimed to investigate the effectiveness of noninvasive positive pressure ventilation (NPPV) in patients with unplanned extubation. MATERIALS AND METHODS: A total of 15 patients (12 male, age: 57 +/- 24 years, APACHE II score: 19 +/- 7) monitored at the medical ICU during the year 2004 who developed unplanned extubation were included in the study. NPPV was tried in all of them following unplanned extubation. Indications for admission to the ICU were as follows: nine patients with pneumonia, three with status epilepticus, one with gastrointestinal bleeding, one with cardiogenic pulmonary edema and one with diffuse alveolar bleeding. RESULTS: Eleven of the patients (74%) were at the weaning period at the time of unplanned extubation. Among these 11 patients, NPPV was successful in 10 (91%) and only one (9%) was reintubated due to the failure of NPPV. The remaining four patients (26%) had pneumonia and none of them were at the weaning period at the time of extubation, but their requirement for mechanical ventilation was gradually decreasing. Unfortunately, an NPPV attempt for 6-8 h failed and these patients were reintubated. CONCLUSIONS: Patients with unplanned extubation before the weaning criteria are met should be intubated immediately. On the other hand, when extubation develops during the weaning period, NPPV may be an alternative. The present study was conducted with a small number of patients, and larger studies on the effectiveness of NPPV in unplanned extubation are warranted for firm conclusions. PMID- 19561918 TI - A neglected problem of developing countries: Noncystic fibrosis bronchiectasis. AB - BACKGROUND: Bronchiectasis has been defined as the abnormal and permanent dilation of the bronchi. It is still an important problem in many developing countries. AIM: The aim of this study was to identify the chacteristics and underlying etiology of children followed with the diagnosis of noncystic fibrosis bronchiectasis. MATERIALS AND METHODS: Children with bronchiectasis confirmed with high-resolution computed tomography were enrolled into the study. The data of the patients, including symptoms of the disease, age at the onset of symptoms, findings of physical examination, labrotory investigations performed in order to identify the etiology of bronchiectasis, etiology of bronchiectasis if found, radiologic findings and treatment modalities were noted. RESULTS: Sixty-six children between 1 and 17 years were included in the study retrospectively. Forty four of them were males (66.7%) and 22 (33.3%) were females. The most common presenting symptoms were cough (100%) and sputum expectoration (50%). An underlying etiology was identified in 44 (66.7%) of the study subjects. The four most common underlying causes were found as infections (21.2%), asthma (16.7%), aspiration syndromes and/or gastroesophageal reflux disease (9.1%) and immunodeficiency syndromes (7.6%), respectively. CONCLUSION: Identifying an underlying etiology will have a significant effect on the management of noncystic fibrosis bronchiectasis. Defining the cause of bronchiectasis may also decrease its incidence, progression and complications. PMID- 19561919 TI - Rates of thoracic trauma and mortality due to accidents in Brazil. AB - AIM: To report on the causes of trauma, indexes of trauma, and mortality related to thoracic trauma in one region of Brazil. MATERIALS AND METHODS: This prospective study was performed at the Regional Trauma Center in Sao Jose do Rio Preto over a 1-year period, from 1(st) July 2004 to 30(th) June 2005. We included all patients attending the center's emergency room with thoracic trauma and an anatomic injury scale (AIS) >= 2. We collected data using a protocol completed on arrival in hospital utilizing the AIS. We studied the types of accidents as well as the mortality and the AIS scores. Prevalence rates were calculated and the paired t-test and logistic regression were employed for the statistical analysis. RESULTS: There were a total of 373 casualties with AIS >= 2 and there were 45 (12%) deaths. The causes of thoracic trauma among the 373 casualties were as follows: 91 (24.4%) car crashes, 75 (20.1%) falls, 46 (12.3%) motorbike accidents, 40 (10.7%) stabbings, 22 (5.9%) accidents involving pedestrians, 21 (5.6%) bicycle accidents, 17 (4.6%) shootings, and 54 (14.5%) other types of accident. The severity of the injuries was classified according to the AIS: 224 (60%) were grade 2, 101 (27%) were grade 3, 27 (7.2%) were grade 4, 18 (4.9%) were grade 5, and 3 were (0.8%) grade 6. With respect to thoracic trauma, pedestrians involved in accidents and victims of shootings had mortality rates that were significantly higher than that of those involved in other types of accidents. CONCLUSION: Road accidents are the main cause of thoracic injury, with accidents involving pedestrians and shootings being associated with a greater death rate. PMID- 19561920 TI - Hemorrhagic pleural effusion secondary to sarcoidosis: A brief review. AB - Pleural effusion is considered to be a rare manifestation of pulmonary sarcoidosis, but hemorrhagic effusion secondary to it is a very uncommon clinical presentation. This case is reported due to the rare manifestation in pulmonary sarcoidosis presenting clinically as hemorrhagic pleural effusion. PMID- 19561921 TI - Skin lesions in a 30-year-old male having smear-positive pulmonary tuberculosis. PMID- 19561924 TI - Respiratory tract infection during Hajj. AB - Respiratory tract infection during Hajj (pilgrimage to Mecca) is a common illness, and it is responsible for most of the hospital admissions. Influenza virus is the leading cause of upper respiratory tract infection during Hajj, and pneumonia can be serious. Taking into account the close contacts among the pilgrims, as well as the crowding, the potential for transmission of M. tuberculosis is expected to be high. These pilgrims can be a source for spreading infection on their return home. Although vaccination program for influenza is implemented, its efficacy is uncertain in this religious season. Future studies should concentrate on prevention and mitigation of these infections. PMID- 19561925 TI - Lung function changes and complications after lobectomy for lung cancer in septuagenarians. AB - BACKGROUND: In septuagenarians, lobectomy is the preferable operation, with lower morbidity than for pneumonectomy. However, the 1-year impact of lobectomy on lung function has not been well studied in elderly patients. MATERIALS AND METHODS: Retrospective study including 30 patients 70 years or older (study group), 25 patients with chronic obstructive pulmonary disease (COPD) under 70 years (control group 1), and 22 patients under 70 years with normal lung function (control group 2) operated for lung cancer in a 2-year period. The study and control groups were compared related to lung function changes after lobectomy, operative morbidity, and mortality. RESULTS: Postoperative lung function changes in the elderly followed the similar trend as in patients with COPD. There were no significant differences between these two groups related to changes in forced expiratory volume in the first second (FEV1) and vital capacity (VC). Unlike that, the pattern of the lung function changes in the elderly was significantly different compared with patients with normal lung function. The mean postoperative decrease in FEV1 was 14.16% in the elderly, compared with a 29.23% decrease in patients with normal lung function (P < 0.05). In the study and control groups, no patients died within the first 30 postoperative days. The operative morbidity in the elderly group was significantly lower than in patients with COPD (23.3% vs. 60%). CONCLUSIONS: The lung function changes after lobectomy in the elderly are similar to those in patients with COPD. The explanation for such a finding needs further investigation. Despite a high proportion of concomitant diseases, the age itself does not carry a prohibitively high risk of operative mortality and morbidity. PMID- 19561923 TI - Obesity hypoventilation syndrome. AB - Obesity is becoming a major medical concern in several parts of the world, with huge economic impacts on health- care systems, resulting mainly from increased cardiovascular risks. At the same time, obesity leads to a number of sleep disordered breathing patterns like obstructive sleep apnea and obesity hypoventilation syndrome (OHS), leading to increased morbidity and mortality with reduced quality of life. OHS is distinct from other sleep- related breathing disorders although overlap may exist. OHS patients may have obstructive sleep apnea/hypopnea with hypercapnia and sleep hypoventilation, or an isolated sleep hypoventilation. Despite its major impact on health, this disorder is under recognized and under-diagnosed. Available management options include aggressive weight reduction, oxygen therapy and using positive airway pressure techniques. In this review, we will go over the epidemiology, pathophysiology, presentation and diagnosis and management of OHS. PMID- 19561926 TI - The six-minute walk test in patients with pulmonary sarcoidosis. AB - BACKGROUND: The 6-min walk test (6MWT) is a useful tool to assess prognosis and functional impairment in various pulmonary diseases. AIMS: To evaluate functional capacity during various stages of pulmonary sarcoidosis and develop a scoring system clinical radiological physiological score (CRP) that can potentially be used to assess the functional status among patients with sarcoidosis. MATERIALS AND METHODS: We performed a retrospective study on 26 patients diagnosed with pulmonary sarcoidosis from 2001 to 2007. All patients completed the 6MWT. The parameters assessed during the test included spirometry, arterial blood gas, 6 min walk distance (6MWD), Borg dyspnea score, and initial and end oxygen saturation. RESULTS: Females covered a significantly shorter distance than males (343 m (223-389) vs. 416.5 m (352-500); P < 0.0001). In addition, females had a significantly lower SpO2 at the end of the 6MWT than males (90.5 (61-99) vs. 96 (75-98); P < 0.03). The 6MWD was inversely correlated with the final Borg score (rho = -0.603, P = 0.004) and the CRP score (rho = -0.364, P = 0.047) and positively correlated with forced expiratory volume in 1 s (FEV1) % ( rho = 0.524, P = 0.006) and forced vital capacity (FVC) % (rho = 0.407, P = 0.039). CONCLUSIONS: Female gender, FEV1%, final Borg score, FVC%, CRP score, and SpO2 at the end of the 6MWT are associated with reduced 6MWD. It appears that Saudi patients diagnosed with sarcoidosis have a markedly reduced walking distance compared with other races. The effect of race and ethnicity and the utility of the CRP score as a potential marker to assess functional status require further exploration. PMID- 19561927 TI - Exhaled nitric oxide in stable chronic obstructive pulmonary disease. AB - STUDY OBJECTIVE: The objective of the study was to test the hypothesis that fraction of exhaled nitric oxide (FENO) is elevated in nonsmoking subjects with stable chronic obstructive pulmonary disease (COPD) and compare it with the results in patients with asthma and a control population. DESIGN: Cross-sectional study. MATERIALS AND METHODS: Pulmonology Clinic at a University Hospital. Twenty five control subjects, 25 steroid naive asthmatics and 14 COPD patients were studied. All the patients were nonsmokers and stable at the time of the study. All subjects completed a questionnaire and underwent spirometry. Exhaled nitric oxide was measured online by chemiluminescence, using single-breath technique. RESULTS: All the study subjects were males. Subjects with stable COPD had significantly higher values of FENO than controls (56.54+/-28.01 vs 22.00+/-6.69; P=0.0001) but lower than the subjects with asthma (56.54+/-28.01 vs 84.78+/-39.32 P=0.0285).The FENO values in COPD subjects were inversely related to the FEV/1FVC ratio. There was a significant overlap between the FENO values in COPD and the control subjects. CONCLUSION: There is a significant elevation in FENO in patients with stable COPD, but the elevation is less than in asthmatic subjects. Its value in clinical practice may be limited by the significant overlap with control subjects. PMID- 19561928 TI - The effect of age on the outcome of esophageal cancer surgery. AB - BACKGROUND: Surgery is still the best way for treatment of esophageal cancer. The increase in life expectancy and the rising incidence of esophageal tumors have led to a great number of elderly candidates for complex surgery. The purpose of this study was to evaluate the effects of advanced age (70 years or more) on the surgical outcome of esophagectomy for esophageal cancer at a single high-volume center. MATERIALS AND METHODS: Between January 2000 and April 2006, 480 cases with esophageal cancer underwent esophagectomy in the referral cancer institute. One hundred sixty-five patients in the elderly group (70 years old or more) were compared with 315 patients in the younger group (< 70 years). All in-hospital morbidity and mortality were studied. RESULTS: The range of age was 38-84 years, with a mean of 58.7. The mean age of the elderly and younger groups was 74 and 53.2, respectively. In the younger group, 70 patients (22.2%) and in the elderly group, 39 patients (23.6%) were complicated (P 0.72).The most common complications in the two groups were pulmonary complications (9.8% in younger and 10.3% in elderly) (P 0.87). Rates of anastomotic leakage and cardiac complications were also similar between the two groups. Hospital mortality rates in younger and elderly patients were 2.8% and 3%, respectively. There was no significant difference between the two groups in morbidities and mortality (P value > 0.05). CONCLUSIONS: With increased experience and care, the outcomes of esophagectomy in elderly patients are comparable to young patients. Advanced age alone is not a contraindication for esophagectomy. PMID- 19561929 TI - Reading chest radiographs in the critically ill (Part I): Normal chest radiographic appearance, instrumentation and complications from instrumentation. PMID- 19561930 TI - A 70 year old male with difficulty in breathing. PMID- 19561931 TI - Prevalence of chronic obstructive pulmonary disease in patients with catheter diagnosed coronary artery disease. PMID- 19561932 TI - Can we change the way we look at BCG vaccine? PMID- 19561934 TI - What's new in emergencies, trauma, and shock? Role of simulation and ultrasound in acute care. PMID- 19561935 TI - The "state of the nation" in trauma critical care: Where are we? PMID- 19561936 TI - Assessing the utility of ultrasound in Liberia. AB - Sub-Saharan Africa has sparse imaging capacity, and data on ultrasound (US) use is limited. We collected prospective data on consecutive patients undergoing US to assess disease spectrum and US utility in Liberia. A total of 102 patients were prospectively enrolled. Average age was 33 years (0-84), 80% were female. US indications were: 53% Obstetrics/Gynecology (OB/GYN) (24% gynecologic, 17% second/third trimester, 12% first trimester), 14% hepatobiliary, 10% intraperitoneal/intrathoracic fluid, 8% cardiac, 5% focused assessment of sonography in trauma, and 4% renal. US changed management in 62% of cases. Greatest impact was in first trimester OB (86%), FAST (83%), ECHO (80%), and second/third trimester OB (77%). US changed management in 47% of right upper quadrant and 33% of gynecologic studies. Curvilinear probe addressed over 80% of need. The primary role for US in developing countries is in management of obstetrics, with a secondary role for traumatic and a-traumatic abdominal processes. Most needs can be met with the curvilinear probe. Training should begin with obstetrics and should be a primary focus for curriculum. PMID- 19561937 TI - Introduction of hi-fidelity simulation techniques as an ideal teaching tool for upcoming emergency medicine and trauma residency programs in India. AB - Emergency medicine (EM) residency programs are a new concept to India. As these programs develop in India the need for effective teaching tools for skills education will rise. A high fidelity simulation workshop was conducted with a intent to expose current residents posted in emergency departments (EDs) to the concept of simulation technology. The participants were subjected to scenarios which tested their core competencies, medical knowledge, and procedural skills using simulation technology. 50 residents were tested over 5 days and an overall satisfaction score and personal comments were assessed to rate the performance of this study. A pre- and post simulation survey was done. Results showed that participants felt that their understanding of communication of expectations increased from 38% fair or good to 76% very good or best. The frequency in which they thought they would ask for help increased from 36% fair or good to 88% very good or best. It was found that students had increased their confidence to challenge a questionable order from a superior from 48% occasionally or half of the time to 76% who would do it the majority of the time or always. In the post survey, 80% would the majority of the time or always admit that they did not know something from 46% who stated they would only do it occasionally or half of the time. We concluded that simulation as a tool for teaching unknown and stressful conditions of ED naturally pair. Resident core competencies can be taught and evaluated more effectively in the simulation lab in a controlled, safe, and collegial manner. PMID- 19561938 TI - Ultrasound diagnosis of traumatic pneumothorax. PMID- 19561939 TI - Leptospirosis: The "mysterious" mimic. AB - Leptospirosis is a potentially fatal bacterial disease that can display a wide array of clinical presentations thus mimicking better-known illnesses. Although, leptospirosis is primarily a zoonotic disease, it frequently inflicts severe illness and death on communities around the globe. A comprehensive overview of the disease in wake of the 2006 outbreaks in India is hereby presented and discussed. PMID- 19561940 TI - Principles of diagnosis and management of traumatic pneumothorax. AB - Presence of air and fluid with in the chest might have been documented as early as Fifth Century B.C. by a physician in ancient Greece, who practiced the so called Hippocratic succession of the chest. This is due to a development of communication between intrapulmonary air space and pleural space, or through the chest wall between the atmosphere and pleural space. Air enters the pleural space until the pressure gradient is eliminated or the communication is closed. Increasing incidence of road traffic accidents, increasing awareness of healthcare leading to more advanced diagnostic procedures, and increasing number of admissions in intensive care units are responsible for traumatic (noniatrogenic and iatrogenic) pneumothorax. Clinical spectrum of pneumothorax varies from asymptomatic patient to life-threatening situations. Diagnosis is usually made by clinical examination. Simple erect chest radiograph is sufficient though; many investigations are useful in accessing the future line of action. However, in certain life-threatening conditions obtaining imaging studies can causes an unnecessary and potential lethal delay in treatment. PMID- 19561941 TI - Policies and programs for road safety in developing India. PMID- 19561942 TI - An unusual presentation of Bell's palsy: A case report and review of literature. AB - In clinical medicine there may be times when clinical conditions manifest differently both when they present individually or concomitantly. Such scenarios warrant a broader differential diagnosis with thorough investigations. We present one such case of a patient of Bell's palsy with unexplained eye pain on the ipsilateral side. The patient had a chronic retinal detachment which became worse due to the concomitant Bell's palsy. PMID- 19561943 TI - Biliary peritonitis due to "fallen" hydatid cyst after abdominal trauma. AB - Hepatic hydatid cysts may cause serious complications. Intraperitoneal rupture of hepatic hydatid cyst is rarely seen and the prognosis can be fatal. By experience, we know that it might be difficult to diagnose an unruptured cyst expulsed into the peritoneal cavity. In this report, we present the case of a 54 year-old man with an intraperitoneal cystic mass of 10 cm of diameter which had extruded out from the liver due to a blunt abdominal trauma. PMID- 19561945 TI - iPhone now. PMID- 19561944 TI - A 19-year-old male with palpitations. AB - A 19-year-old male presented to the emergency department (ED) following intermittent episodes of palpitations. Classical "epsilon waves" noted on his initial electrocardiogram prompted an evaluation for arrhythmogenic right ventricular dysplasia (ARVD). The diagnosis was confirmed with magnetic resonance imaging of the heart and stress test. A prompt recognition and management of this condition in the ED helped prevent significant mortality that may be associated with ARVD. PMID- 19561946 TI - What's new in emergencies, trauma and shock? Publishing clinical trials in JETS from countries around the globe. PMID- 19561947 TI - Use of antiemetics in children with acute gastroenteritis: Are they safe and effective? AB - The use of antiemetics is a controversial topic in treatment of pediatric gastroenteritis. Although not recommended by the American Academy of Pediatrics, antiemetics are commonly prescribed by physicians. A review of the literature shows side effects of promethazine, prochlorperazine, and metoclopramide are common and potentially dangerous. Ondansetron has recently been studied as an adjunct to oral rehydration therapy in treatment of acute gastroenteritis with mild to moderate dehydration. Although studies are limited, early research suggests the medication is safe when used in a single dose and can be effective to prevent vomiting, the need for intravenous fluids, and hospital admission. PMID- 19561948 TI - Vascular endothelial growth factor and acute mountain sickness. AB - STUDY OBJECTIVE: Despite causing significant morbidity throughout the mountainous regions of the world, the pathophysiology of acute mountain sickness (AMS) remains poorly understood. This study aims to improve the understanding of altitude illness by determining if vascular endothelial growth factor (VEGF) plays a role in the development of AMS. The purpose of this study was to determine if elevated plasma VEGF correlates with increased symptoms of AMS at high altitude. PATIENTS AND METHODS: This is a prospective study of a cohort of healthy climbers on Denali (Mount McKinley) in Alaska at 14, 200 feet. Baseline demographics, medications, rates of ascent, and AMS scores were recorded. Pulse oximetry measurements and venous blood samples were obtained. Comparisons were made between mountaineers with and without AMS. RESULTS: Seventy-two climbers were approached for participation in the study; 21 (29%) refused. Of the 51 climbers participating in the study, 14 subjects (27.5%) had symptoms of AMS and 37 subjects (72.5%) were free of symptoms of AMS. Plasma VEGF levels were 79.14 pg/dl (SD: 121.44) and 57.57pg/dl (SD: 102.71) in the AMS and non-AMS groups, respectively. These results were nonsignificant. Similarly, comparison of sex, age, rate of ascent, pulse oximetry values, or history of altitude illness did not reveal significant differences between the AMS and non-AMS groups. CONCLUSION: This study does not provide evidence in support of the theory that plasma VEGF correlates with symptoms of AMS. PMID- 19561949 TI - Management of Ocular Trauma in Emergency (MOTE) Trial: A pilot randomized double blinded trial comparing topical amethocaine with saline in the outpatient management of corneal trauma. AB - BACKGROUND: It is unclear whether local anesthetic eye drops can be safely used for the topical anesthesia of patients with minor corneal injury who are discharged from the emergency department (ED). OBJECTIVES: To assess whether topical 0.4% amethocaine self-administered to a maximum recommended frequency of once every hour for 36-48 h is safe in the management of uncomplicated corneal injury in patients discharged from the ED. PATIENTS AND METHODS: A pilot randomized double-blinded trial comparing topical 0.4% amethocaine with topical normal saline. RESULTS: Forty-seven subjects were recruited, with 22 randomized to receive amethocaine and 25 to receive placebo (normal saline). Baseline characteristics, including corneal injury type, were similar in both groups. There were no significant functional or clinical adverse sequelae in the majority of enrolled patients who could be contacted at 2 weeks (17/22 for amethocaine and 21/25 for placebo). Follow-up for the primary study outcome was suboptimal, with only 7/22 from the amethocaine group and 9/25 from the saline group presenting for 36-48 h review; there was a statistically nonsignificant trend for persistence of the corneal defect in the amethocaine group as compared with the saline group (2/7 and 1/9, respectively). CONCLUSION: Compared with saline drops, amethocaine eye drops are not definitely safe but they are effective for topical analgesia in minor corneal injury. Until further definitive studies, topical nonsteroidal agents or long-lasting artificial tears may be preferred for the topical analgesia of minor corneal injury. Return for corneal re-evaluation will necessarily remain suboptimal in an otherwise self-limiting condition, leading to a bias even if study recruitment is good. PMID- 19561950 TI - On-call emergency workload of a general surgical team. AB - BACKGROUND: To examine the on-call emergency workload of a general surgical team at a tertiary care teaching hospital to guide planning and provision of better surgical services. PATIENTS AND METHODS: During six months period from August to January 2007; all emergency calls attended by general surgical team of Surgical Unit II in Accident and Emergency department (A and E) and in other units of Civil, Hospital Karachi, Pakistan were prospectively recorded. Data recorded includes timing of call, diagnosis, operation performed and outcome apart from demography. RESULTS: Total 456 patients (326 males and 130 females) were attended by on-call general surgery team during 30 emergency days. Most of the calls, 191 (41.9%) were received from 8 am to 5 pm. 224 (49.1%) calls were of abdominal pain, with acute appendicitis being the most common specific pathology in 41 (9.0%) patients. Total 73 (16.0%) calls were received for trauma. Total 131 (28.7%) patients were admitted in the surgical unit for urgent operation or observation while 212 (46.5%) patients were discharged from A and E. 92 (20.1%) patients were referred to other units with medical referral accounts for 45 (9.8%) patients. Total 104 (22.8%) emergency surgeries were done and the most common procedure performed was appendicectomy in 34 (32.7%) patients. CONCLUSION: Major workload of on-call surgical emergency team is dealing with the acute conditions of abdomen. However, significant proportion of patients are suffering from other conditions including trauma that require a holistic approach to care and a wide range of skills and experience. These results have important implications in future healthcare planning and for the better training of general surgical residents. PMID- 19561951 TI - Using continuous renal replacement therapy to manage patients of shock and acute renal failure. AB - BACKGROUND: The incidence of acute renal failure (ARF) in the hospital setting is increasing. It portends excessive morbidity and mortality and a considerable burden on hospital resources. Extracorporeal therapies show promise in the management of patients with shock and ARF. It is said that the potential of such therapy goes beyond just providing renal support. The aim of our study was to analyze the clinical setting and outcomes of critically ill ARF patients managed with continuous renal replacement therapy (CRRT). PATIENTS AND METHODS: Ours was a retrospective study of 50 patients treated between January 2004 and November 2005. These 50 patients were in clinical shock and had concomitant ARF. All of these patients underwent CVVHDF (continuous veno-venous hemodiafiltration) in the intensive care unit. For the purpose of this study, shock was defined as systolic BP < 100 mm Hg in spite of administration of one or more inotropic agents. SOFA (Sequential Organ Failure Assessment) score before initiation of dialysis support was recorded in all cases. CVVHDF was performed using the Diapact((R)) (Braun) CRRT machine. The vascular access used was as follows: femoral in 32, internal jugular in 8, arteriovenous fistula (AVF) in 4, and subclavian in 6 patients. We used 0.9% or 0.45% (half-normal) saline as a prefilter replacement, with addition of 10% calcium gluconate, magnesium sulphate, sodium bicarbonate, and potassium chloride in separate units, while maintaining careful monitoring of electrolytes. Anticoagulation of the extracorporeal circuit was achieved with systemic heparin in 26 patients; frequent saline flushes were used in the other 24 patients. RESULTS: Of the 50 patients studied, 29 were males and 21 females (1.4:1). The average age was 52.88 years (range: 20-75 years). Causes of ARF included sepsis in 24 (48%), hemodynamically mediated renal failure (HMRF) in 18 (36%), and acute over chronic kidney disease in 8 (16%) patients. The overall mortality was 74%. The average SOFA score was 14.31. The variables influencing mortality on multivariate analysis were: age [odds ratio (OR):1.65; 95% CI: 1.35 to 1.92; P = 0.04], serum creatinine (OR:1.68; 95% CI: 1.44 to 1.86; P = 0.03), and serum bicarbonate (OR: 0.76; 95% CI: 0.55 to 0.94; P = 0.01). On univariate analysis the SOFA score was found to be a useful predictor of mortality. CONCLUSIONS: Despite advances in treating critically ill patients with newer extracorporeal therapies, mortality is dismally high. Multiorgan dysfunction adversely affects outcome of CRRT. Older age, level of azotemia, and severity of metabolic acidosis are important predictors of adverse outcome. PMID- 19561952 TI - Injuries, negative consequences, and risk behaviors among both injured and uninjured emergency department patients who report using alcohol and marijuana. AB - BACKGROUND: Brief intervention (BI) to reduce hazardous drinking and negative consequences such as injury has been effective when given in the emergency department (ED). The effectiveness and effect of BI has varied between injured and uninjured ED patients. This study compares injured and uninjured ED patients who admit to alcohol and marijuana use to determine their need and their readiness for BI. PATIENTS AND METHODS: Participants volunteered to enter a randomized controlled trial of BI to reduce hazardous alcohol and marijuana use. Adult ED patients who had had alcohol in the last month and smoked marijuana in the last year were recruited. Those patients who were admitted to hospital, were under police custody, or were seeking treatment for substance use or psychiatric disorder were excluded. Research assistants interviewed participants using a validated questionnaire. Data were analyzed using SAS (version 9.1). Binominal tests of proportions, t-test analyses, and transformations were conducted as appropriate. RESULTS: Injured (n = 249) and uninjured (n = 266) study participants reported very high, statistically equivalent (P > 0.05), rates of binge drinking (4-5 days/month), marijuana use (13 days/month), driving under the influence of marijuana or alcohol (>49% in the last 3 months), injury (>83% in the last year), and other negative consequences (>64% in the last 3 months) prior to their ED visit. These behaviors and the consequences demonstrate a need for change. Both injured and uninjured subjects were ready to change (>56%) and confident they could change (>91%) alcohol and marijuana use. DISCUSSION: ED patients who admit to alcohol and marijuana use also use other hazardous substances and participate in high-risk behaviors. In both injured and uninjured patients who admit using alcohol and marijuana, the ED visit is an opportunity to deliver BI to reduce alcohol and marijuana use and associated risk behaviors and the subsequent injury and negative consequences. Given their risk behaviors and experience of negative consequences, members of both injured and uninjured groups have an equal need for BI. Fortunately, in both groups, a high number of members express motivation to change. PMID- 19561953 TI - Emergency management of fat embolism syndrome. AB - Fat emboli occur in all patients with long-bone fractures, but only few patients develop systemic dysfunction, particularly the triad of skin, brain, and lung dysfunction known as the fat embolism syndrome (FES). Here we review the FES literature under different subheadings.The incidence of FES varies from 1-29%. The etiology may be traumatic or, rarely, nontraumatic. Various factors increase the incidence of FES. Mechanical and biochemical theories have been proposed for the pathophysiology of FES. The clinical manifestations include respiratory and cerebral dysfunction and a petechial rash. Diagnosis of FES is difficult. The other causes for the above-mentioned organ dysfunction have to be excluded. The clinical criteria along with imaging studies help in diagnosis. FES can be detected early by continuous pulse oximetry in high-risk patients. Treatment of FES is essentially supportive. Medications, including steroids, heparin, alcohol, and dextran, have been found to be ineffective. PMID- 19561954 TI - Cardiac arrest and pregnancy. AB - Cardiopulmonary arrest in pregnancy is rare occurring in 1 in 30,000 pregnancies. When it does occur, it is important for a clinician to be familiar with the features peculiar to the pregnant state. Knowledge of the anatomic and physiologic changes of pregnancy is helpful in the treatment and diagnosis. Although the main focus should be on the mother, it should not be forgotten that there is another potential life at stake. Resuscitation of the mother is performed in the same manner as in any other patient, except for a few minor adjustments because of the changes of pregnancy. The specialties of obstetrics and neonatology should be involved early in the process to ensure appropriate treatment of both mother and the newborn. This article will explore the changes that occur in pregnancy and their impact on treatment. The common causes of maternal cardiac arrest will be discussed briefly. PMID- 19561955 TI - Cutaneous absorption of Oleander: Fact or fiction. AB - Cardiac conduction disorders following oral ingestion of Oleander plant materials were documented earlier. Transcutaneous absorption of yellow oleander (Thevetia peruviana) leaf extract applied over non intact skin (raw wound) resulting in reversible cardiac conduction disorder observed in four healthy males who were free from any other systemic or electrolyte or metabolic disorders or exposure to pesticide or toxins is reported for the first time. Their hematological, biochemical, clinical, and echocardiogram status were within normal limits and free of any abnormalities. One among the four, presented for weakness and breathlessness (class II). He had bradycardia with Mobitz II block and hypotension without any other demonstrable localizing signs. The other three were identified in the community and without any symptoms. However, their ECG revealed bradycardia with Mobitz I block in two and complete heart block in the other. All of the four recovered well without any untoward events. Hence, it is suggested that physicians and practitioners have to elicit history and route of administration of unconventional therapy, whenever they are confronted with clinical challenges and during medical emergencies before embarking final decision. PMID- 19561956 TI - Simultaneous acute appendicitis and ectopic pregnancy. AB - The acute abdomen in pregnancy is a surgical emergency. Ectopic pregnancy and appendicitis are two causes of acute abdomen in pregnancy. Difficulties in correctly identifying the cause of the pain can be hazardous to the patient and care needs to be taken in obtaining a prompt and accurate diagnosis enabling the most appropriate management. The case presented here underlies the pathogenesis of the simultaneous existence of these two conditions in a patient. PMID- 19561957 TI - Chronic subdural hematoma leading to fatal cavernous sinus thrombosis. AB - Presented is a case of cavernous sinus thrombosis in a young female with fatal outcome. There were not any septic focus, no history of head trauma, no relation with pregnancy. Computed tomography scan of brain showed chronic subdural hematoma. An attempt is made to correlate the aetiopathology with the clinical features of this rare case presentation. PMID- 19561958 TI - Emergency intubation using a light wand in patients with facial trauma. AB - Airway management in the operating room is the responsibility of anesthesiologists, although a variety of personnel may be responsible for airway management outside the operating room. Emergency department physicians are prominently involved in airway management in the emergency room both independently and with anesthesiologists. Airway management in trauma patients remains the domain of anesthesiologists. An 18-year old male patient was brought to our emergency room after an alleged history of suicidal attempt with gunshot under the chin. He was scheduled to undergo emergency tracheotomy, debridement, and closure of facial laceration under general anaesthesia, presenting a challenge for. He had to undergo emergency tracheotomy, debridement, and closure of facial lacerations under general anesthesia. The injuries made the patient's airway management a complex issue. We present the use of the light wand to manage the difficult airway of this patient with complex facial trauma. PMID- 19561960 TI - Early tracheostomy in severe head injuries at a rural center. PMID- 19561959 TI - Emergency battlefield cricothyrotomy complicated by tube occlusion. AB - Emergency cricothyrotomy is a technique used to secure an otherwise compromised or inaccessible airway and has been recommended for use in the battlefield under certain circumstances. This case reports an acute complication of emergency cricothyrotomy. An Iraqi soldier, injured in an improvised explosive device blast received an emergency battlefield cricothyrotomy. At the Combat Support Hospital, the patient became more difficult to ventilate and was taken to the operating room for tracheostomy. The cricothyrotomy tube was found to be occluded with blood. PMID- 19561961 TI - Safe involvement of first year residents in patients care. PMID- 19561962 TI - What's new in emergencies, trauma, and shock? JETS policy for publishing animal studies. PMID- 19561963 TI - Role of pyruvate dehydrogenase complex in traumatic brain injury and Measurement of pyruvate dehydrogenase enzyme by dipstick test. AB - OBJECTIVES: The present study was designed to investigate the role of a mitochondrial enzyme pyruvate dehydrogenase (PDH) on the severity of brain injury, and the effects of pyruvate treatment in rats with traumatic brain injury (TBI). METHODS: We examined rats subjected to closed head injury using a fluid percussion device, and treated with sodium pyruvate (antioxidant and substrate for PDH enzyme). At 72 h post injury, blood was analyzed for blood gases, acid base status, total PDH enzyme using a dipstick test and malondialdehyde (MDA) levels as a marker of oxidative stress. Brain homogenates from right hippocampus (injured area) were analyzed for PDH content, and immunostained hippocampus sections were used to determine the severity of gliosis and PDH E1-infinity subunit. RESULTS: Our data demonstrate that TBI causes a significant reduction in PDH enzyme, disrupt-acid-base balance and increase oxidative stress in blood. Also, lower PDH enzyme in blood is related to the increased gliosis and loss of its PDH E1-infinity subunit PDH in brain tissue, and these effects of TBI were prevented by pyruvate treatment. CONCLUSION: Lower PDH enzyme levels in blood are related to the global oxidative stress, increased gliosis in brain, and severity of brain injury following TBI. These effects can be prevented by pyruvate through the protection of PDH enzyme and its subunit E-1. PMID- 19561964 TI - Evaluating the validity of multiple imputation for missing physiological data in the national trauma data bank. AB - BACKGROUND: The National Trauma Data Bank (NTDB) is plagued by the problem of missing physiological data. The Glasgow Coma Scale score, Respiratory Rate and Systolic Blood Pressure are an essential part of risk adjustment strategies for trauma system evaluation and clinical research. Missing data on these variables may compromise the feasibility and the validity of trauma group comparisons. AIMS: To evaluate the validity of Multiple Imputation (MI) for completing missing physiological data in the National Trauma Data Bank (NTDB), by assessing the impact of MI on 1) frequency distributions, 2) associations with mortality, and 3) risk adjustment. METHODS: Analyses were based on 170,956 NTDB observations with complete physiological data (observed data set). Missing physiological data were artificially imposed on this data set and then imputed using MI (MI data set). To assess the impact of MI on risk adjustment, 100 pairs of hospitals were randomly selected with replacement and compared using adjusted Odds Ratios (OR) of mortality. OR generated by the observed data set were then compared to those generated by the MI data set. RESULTS: Frequency distributions and associations with mortality were preserved following MI. The median absolute difference between adjusted OR of mortality generated by the observed data set and by the MI data set was 3.6% (inter-quartile range: 2.4%-6.1%). CONCLUSIONS: This study suggests that, provided it is implemented with care, MI of missing physiological data in the NTDB leads to valid frequency distributions, preserves associations with mortality, and does not compromise risk adjustment in inter-hospital comparisons of mortality. PMID- 19561965 TI - Medical errors and consequent adverse events in critically ill surgical patients in a tertiary care teaching hospital in Delhi. AB - BACKGROUND: Medical errors and adverse events (AE), though common worldwide, have never been studied in India. We believe that though common these are under reported. AIM: The aim of this study was to study medical errors and consequent AE in patients presenting with trauma and bowel perforation peritonitis. METHODS: Five hundred and eighty-six consecutive patients with trauma or peritonitis, presenting to surgery emergency of UCMS-GTBH, were prospectively studied using review form (RF) 1 and 2. AE was defined as an outcome not expected to be part of the illness. RF 1 was filled for all and indicated if AE was present or not. RF2 was filled when RF 1 indicated presence of AE; it further confirmed the occurrence of AE and pointed to the type of medical error and resultant disability. All results were expressed as percentage. RESULTS: There were 500 (85%) males. Mean age of the patients was 31 years. There were 332 patients with peritonitis and 254 with trauma. AE and its consequences were present in 185 (31.5%) and 183 (31.2%) patients, respectively. Consequences were as follows: disability - 157 (85%), increased hospital stay and/or increased visits in the OPD - 28 (15.3%) and both-101 (55.2%) patients. Disabilities were: death - 62 (40%), temporary disability - 90 (58%) and permanent disability - 05 (3.1%) patients. AE in 133 (71.8%) patients was definitely (level of confidence 6) due to error in healthcare management. All AE were considered preventable. Error of omission accounted for AE in 122 (65.9%) patients. System and operative errors were the commonest, 84.3% and 82.7%, respectively. One hundred and sixty-seven (90%) patients had multiple errors. CONCLUSIONS: The study proves that medical errors and AE are a serious problem in our set-up and calls for immediate system improvement. PMID- 19561966 TI - Simulation in a high stakes clinical performance exam. AB - BACKGROUND: Hi-fidelity simulation is becoming accepted as a teaching tool for medical providers. Advanced simulations allow educators to test difficult clinical scenarios. The goal of this study was to test the diagnostic and treatment skills of a third-year medical student faced with a simulated patient having evidence of a stable pneumothorax. Students are then expected to evaluate the teaching simulation in comparison to traditional methods. METHODS: The case was one of a 12 cases in the "high stakes" Clinical Performance Exam. The patient with evidence of a stable pneumothorax was chosen to evaluate both diagnostic abilities and decision making in therapeutic options. Students were assessed using a university-wide standardized checklist: diagnosis, management, and interaction with the simulator. Immediately following the simulation, the students evaluated the experience. RESULTS: The exam was given to 117 students. The correct diagnosis was made by 115/117 (98%). Treatment was considered acceptable in a majority of students, Send patient to the Emergency Department 77%, Oxygen 26% and Analgesia 39%. The follow-up survey completed by 78% of the students revealed the students felt comfortable with the simulators, but had concerns about the exam. Students liked the simulator as an educational tool 88% of the time. CONCLUSIONS: Simulation was used in a year-end exam and majority of students chose the correct diagnosis and treatment plan. It was also found that a significant percentage of students performed an unnecessary and potentially harmful procedure. The survey revealed that students were concerned about distractions and realism, but overall expressed desire for more education using simulation. PMID- 19561968 TI - Connecting care competencies and culture during disasters. AB - Connecting care Competencies and Culture are core fundamentals in responding to disasters. Thick coordination between professionals, communities and agencies in different geographical areas is crucial to the happening of appropriate preparedness and thus efficient response and mitigation of a disaster. In the next few articles, we present diverse examples related to the preparedness and recovery process to adverse disasters across the globe. PMID- 19561967 TI - Characteristics of associated craniofacial trauma in patients with head injuries: An experience with 100 cases. AB - BACKGROUND: Facial fractures and concomitant cranial injuries carry the significant potential for mortality and neurological morbidity mainly in young adults. AIMS AND OBJECTIVES: To analyze the characteristics of head injuries and associated facial injuries, the management options and outcome following cranio facial trauma. METHODS: This retrospective review was performed at Justice K. S. Hegde Charitable Hospital, and associated A. B. Shetty Memorial Institute of Dental sciences, Deralakatte, Mangalore. Following Ethical Committee approval, hospital charts and radiographs of 100 consecutive patients of cranio-facial trauma managed at the Department of Oral and Maxillofacial Surgery and Neurosurgery between January 2004 and December 2004 were reviewed. RESULTS: Majority of the patients were in the 2nd to 4th decade (79%) with a male to female ratio of -8.09:1. Road traffic accidents were the common cause of craniofacial trauma in present study (54%) followed by fall from height (30%). Loss of consciousness was the most common clinical symptom (62%) followed by headache (33%). Zygoma was the most commonly fractured facial bone 48.2% (alone 21.2%, in combination 27.2%). Majority of patients had mild head injury and managed conservatively in present series. Causes of surgical intervention for intracranial lesions were compound depressed fracture, contusion and intracranial hematoma. Operative indications for facial fractures were displaced facial bone fractures. Major causes of mortality were associated systemic injuries. CONCLUSION: Adult males are the most common victims in craniofacial trauma, and road traffic accidents were responsible for the majority. Most of the patients sustained mild head injuries and were managed conservatively. Open reduction and internal fixation with miniplates was used for displaced facial bone fractures. PMID- 19561969 TI - Swine influenza A (H1N1) strikes a potential for global disaster. AB - As of April 25(th)2009, 11.00 AM, eight human cases of swine influenza A virus infection have been identified in the United States in California and Texas. There is also established evidence of similar cases across the United States border in Mexico. Experts from the Centers for Disease Control and Prevention in cooperation with World Health Organization and public health experts from Canada and Mexico are leading an exhaustive investigation to find the source of infection and infected people. We present a profile of this illness from the available literature. PMID- 19561970 TI - Cyclone Nargis and Myanmar: A wake up call. AB - In early May 2008, Cyclone Nargis (CN) tore across the southern coastal regions of Myanmar, pushing a tidal surge through villages and rice paddies. The almost 12 foot wall of water and wind speed of over 200 km/hr killed tens of thousands of people and left hundreds of thousands homeless and vulnerable to injury and disease. Out of the 7.35 million living in the affected townships of Labutta, Bogale, Pyinsalu, Yangon, and many more, approximately 2.4 million were affected. Overall, more than 50 townships were affected by this most devastating cyclone in Asia since 1991. The Delta region, Myanmar's Rice Bowl, was severely damaged. The low-lying villages were submerged. There was widespread destruction of homes, critical infrastructure of the villages, roads, ferries, water, fuel, and electricity supplies. Our team from Singapore (called Team Singapore) reached out to at least 10 different villages during the time we were there. We ran mobile clinics daily at several locations and these operated from warehouses, temples, schools or any make shift buildings. The journey to the remote villages may take between 1 and 2 hours by road or by boat. We also ran mobile clinics at the township hospital, the rural healthcare centers, and an orphanage. PMID- 19561971 TI - Using a joint triage model for multi-hospital response to a mass casualty incident in New York city. AB - This paper defines a specific plan which allows two separate institutions, with different capabilities, to function as a single receiving entity in the event of a mass casualty incident. The street between the two institutions will be closed to traffic and a two-phase process initiated. Arriving ambulances will first be quickly screened to expedite the most critical patients followed by formal triage and directing patients to one of the two facilities. Preparation for this plan requires prior coordination between local authorities and the administrations of both institutions. This plan can serve as a general model for disaster preparedness when two or more institutions with different capabilities are located in close proximity. PMID- 19561972 TI - Health implications of radiological terrorism: Perspectives from Israel. AB - September 11(th) events taught us, members of the medical community, that we need to prepared for the worst. Nuclear terror is no longer science fiction. Radiological weapons of mass terror come in three flavors: The first one is nuclear. Since 1992, there have been six known cases of highly enriched uranium or plutonium being intercepted by authorities as it passed in or out of the former Soviet Union. Constructing a nuclear fission weapon requires high-level expertise, substantial facilities, and lots of money. All three of which would be difficult, although not impossible, for a terrorist group to pull off without state support. However, terrorists could carry out potential mass destruction without sophisticated weaponry by targeting nuclear facilities using conventional bombs or hijacked aircrafts. Terror attacks could also carry out mass panic and radioactive contamination of people and environment by dispersal of radioactive materials with or without the use of conventional explosive devices. Most medical and para-medical personnel are not familiar with CBRN terror and radiation casualties. To lessen the impact of those potential attacks and provide care for the greatest number of potential survivors, the community as a whole - and the medical community in particular - must acquire the knowledge of the various signs and symptoms of exposure to irradiation and radioactive contamination as well as have a planned response once such an attack has occurred. Based on knowledge of radiation hazards, medical emergency planers should analyze the risks of each scenario, offer feasible solutions and translate them into internationally accepted plans that would be simple to carry out once such an attack took place. The planned response should be questioned and tested by drills. Those drills should check the triage, evacuation routes, decontamination posts, evacuation centers and receiving hospitals. It is crucial that the drill will consist of simulated casualties that will follow the evacuation route from point zero to the ED. Knowledge and exercise will reduce terror (fear) from radiation and help the community as a whole better cope with such an event. This article will review the general information of radiation types, their biological damage, clinical appearance and general concepts of nuclear event planning, focusing on medical response and focus on the Israeli perspective. PMID- 19561973 TI - Emergency management of decompensated peripartum cardiomyopathy. AB - Peripartum cardiomyopathy (PPCM) is a rare life-threatening cardiomyopathy of unknown cause that occurs in the peripartum period in previously healthy women.[1] the symptomatic patients should receive standard therapy for heart failure, managed by a multidisciplinary team. The diagnosis of PPCM rests on the echocardiographic identification of new left ventricular systolic dysfunction during a limited period surrounding parturition. Diagnostic criteria include an ejection fraction of less than 45%, fractional shortening of less than 30%, or both, and end-diastolic dimension of greater than 2.7 cm/m(2) body surface-area. This entity presents a diagnostic challenge because many women in the last month of a normal pregnancy experience dyspnea, fatigue, and pedal edema, symptoms identical to early congestive heart failure. There are no specific criteria for differentiating subtle symptoms of heart failure from normal late pregnancy. Therefore, it is important that a high index of suspicion be maintained to identify the rare case of PPCM as general examination showing symptoms of heart failure with pulmonary edema. PPCM remains a diagnosis of exclusion. No additional specific criteria have been identified to allow distinction between a peripartum patient with new onset heart failure and left ventricular systolic dysfunction as PPCM and another form of dilated cardiomyopathy. Therefore, all other causes of dilated cardiomyopathy with heart failure must be systematically excluded before accepting the designation of PPCM. Recent observations from Haiti[2] suggest that a latent form of PPCM without clinical symptoms might exist. The investigators identified four clinically normal postpartum women with asymptomatic systolic dysfunction on echocardiography, who subsequently either developed clinically detectable dilated cardiomyopathy or improved and completely recovered heart function. PMID- 19561974 TI - Hair dye poisoning and the developing world. AB - Hair dye poisoning has been emerging as one of the important causes of intentional self harm in the developing world. Hair dyes contain paraphenylene diamine and a host of other chemicals that can cause rhabdomyolysis, laryngeal edema, severe metabolic acidosis and acute renal failure. Intervention at the right time has been shown to improve the outcome. In this article, we review the various manifestations, clinical features and treatment modalities for hair dye poisoning. PMID- 19561975 TI - Difficult weaning in delayed onset diaphragmatic hernia. AB - Diaphragmatic injuries are relatively rare and result from either blunt or penetrating trauma. Regardless of the mechanism, diagnosis is often missed and high index of suspicion is vital. The clinical signs associated with a diaphragmatic hernia can range from no outward signs to immediately life threatening respiratory compromise. Establishing the clinical diagnosis of diaphragmatic injuries (DI) can be challenging as it is often clinically occult. Accurate diagnosis is critical since missed DI may result in grave sequelae due to herniation and strangulation of displaced intra-abdominal organs. We present a case of polytrauma with rib fracture and delayed appearance of diaphragmatic hernia manifesting as difficult weaning from ventilatory support. PMID- 19561976 TI - Fatal Klebsiella pneumoniae meningitis and concomitant disseminated intravascular coagulation in a patient with diabetes mellitus. AB - Bacterial meningitis remains a major cause of death and long-term neurologic sequelae worldwide. We present a case of fatal Klebsiella pneumoniae meningitis and concomitant disseminated intravascular coagulation (DIC) in a 72-year-old woman with diabetes mellitus (DM). Both blood and cerebrospinal fluid cultures grew Klebsiella pneumoniae. Due to advanced age, newly recognized DM, K. pneumoniae bacteremia, and DIC, the prognosis of our patient was poor. Eight hours after arrival to the emergency department, cardiopulmonary resuscitation was necessary in this patient, but she died despite an early diagnosis and appropriate antibiotic therapy. PMID- 19561977 TI - Retroperitoneal hemorrhagic shock in a patient on warfarin therapy. AB - Oral anticoagulants are an established treatment modality in the prophylaxis of thromboembolic events in various clinical scenarios. Needless to say that, bleeding is a natural adverse effect of this drug. Most of the times bleeding is inconsequential. But nevertheless massive and fatal bleeding can occur occasionally. The case reported here is rare, as the patient presented with massive hemoperitoneum due to mesenteric hemorrhage and hemorrhagic infarction of small bowel necessitating Laparotomy resection. PMID- 19561978 TI - Delayed presentation of shock due to retroperitoneal hemorrhage following a fall. AB - During trauma the abdomen is one region which cannot be ignored. Due to its Complex anatomy it is very important that all the areas in the abdomen be examined both clinically and radiologicaly to rule out any abdominal bleeding as a cause of Hemorrhagic Shock Following Trauma. Our case justifies the above. PMID- 19561979 TI - The four killers of Meckel's diverticulum. PMID- 19561980 TI - Unexpected retraction of distal cut end of flexor pollicis longus tendon. PMID- 19561981 TI - What's new in emergencies, trauma, and shock? Snake envenomation and organophosphate poisoning in the emergency department. PMID- 19561983 TI - Dermal absorption of a dilute aqueous solution of malathion. AB - Malathion is an organophosphate pesticide commonly used on field crops, fruit trees, livestock, agriculture, and for mosquito and medfly control. Aerial applications can result in solubilized malathion in swimming pools and other recreational waters that may come into contact with human skin. To evaluate the human skin absorption of malathion for the assessment of risk associated with human exposures to aqueous solutions, human volunteers were selected and exposed to aqueous solutions of malathion. Participants submerged their arms and hands in twenty liters of dilute malathion solution in either a stagnant or stirred state. The "disappearance method" was applied by measuring malathion concentrations in the water before and after human exposure for various periods of time. No measurable skin absorption was detected in 42% of the participants; the remaining 58% of participants measured minimal absorbed doses of malathion. Analyzing these results through the Hazard Index model for recreational swimmer and bather exposure levels typically measured in contaminated swimming pools and surface waters after bait application indicated that these exposures are an order of magnitude less than a minimal dose known to result in a measurable change in acetylcholinesterase activity. It is concluded that exposure to aqueous malathion in recreational waters following aerial bait applications is not appreciably absorbed, does not result in an effective dose, and therefore is not a public health hazard. PMID- 19561982 TI - Military anti-shock garment: Historical relic or a device with unrealized potential? AB - Military anti-shock trousers represents a medical device which has engendered very divergent views, even up to today. From the time the concept was formulated in 1903 by surgeon George W Crile, there have been significant swings in opinion and evidence. The guidelines, where available, are often kept relatively general and cautious. As a spin-off to the mechanism and technology, several alternative devices have been proposed or developed over the years. This include the auto transfusion torniquet, the non pneumatic anti-shock garment (Life Wrap) and the non inflatable antishock garment, which are discussed in this paper. PMID- 19561984 TI - Are trauma patients better off in a trauma ICU? AB - There is very little data on the value of specialized intensive care unit (ICU) care in the literature. To determine if specialize ICU care for the trauma patient improved outcomes in this patient population. Level I Trauma Center Compared outcomes of trauma patients treated in a surgical trauma ICU (STICU) to those treated in non- trauma ICUs (non-STICU). Retrospective review of trauma registry data. STATISTICAL ANALYSIS: Wilcoxon Rank Test, Fischer's Exact test, logistic regression. There were 1146 STICU patients compared to 1475 non-STICU. In all ISS groups there were more penetrating trauma patients in the STICU (32.54% STICU vs. 18.15% non-STICU, P<0.0001 (ISS< 15)), (21.03% STICU vs. 12.98% non-STICU, P=0.0074 (ISS between 15-25)), and (19.42% STICU vs. 11.35% non-STICU, P=0.0026 (ISS> 25)). All groups had similar lengths of stay. The blunt trauma patients were sicker in the STICU (20.8 ISS +/- 12.2 STICU vs. 19.7 ISS +/- 11.9 non-STICU, P=0.03) yet had similar outcomes to the non-STICU group. Logistic regression identified penetrating trauma and not ICU location as a predictor of mortality. Sicker STICU patients do as well as less injured non-STICU patients. Severely injured patients should be preferentially treated in a STICU where they are better equipped to care for the complex multi-trauma patient. All patients, regardless of location, do well when their management is guided by a surgical critical care team. PMID- 19561985 TI - Clinical profile of venomous snake bites in north Indian Military Hospital. AB - Snakebite is an environmental hazard associated with significant morbidity and mortality. We report a case series of venomous snakebites in a military operational area of north India. Of 33 cases of snake bites presenting to the military hospital, 21 patients were envenomated. The median age of patients was 24 years; all were men. All of the envenomations were neurotoxic in nature. Abdominal pain (91%), headache (86%), dysphagia (86%), ptosis (77%), diplopia (72%), blurred vision (72%), dyspnea (67%), and vomiting (62%) were the predominant clinical presentation. Polyvalent AntiSnakeVenom (ASV) [mean 180 ml; range 90-320 ml] was given to all patients with systemic manifestations, and repeated as needed. Eleven (52%) patients received neostigmine with glycopyrrolate to counter cholinergic effects. Two patients were given ventilatory support. The average time of recovery from envenomation was 16 hours after administration of ASV. All patients recovered without sequelae. Soldiers during military exercise are vulnerable to snakebites. Neurotoxic snakebites predominate in our study and usually present with autonomic features along with headache, abdominal pain, ptosis, diplopia and dysphasia. Preventive measures to minimize snake bites and planned treatment regimens should be emphasized among medical and military personnel deployed in the field operations. PMID- 19561986 TI - Alcohol, burn injury, and the intestine. AB - A significant number of burn and other traumatic injuries are reported to occur under the influence of alcohol (EtOH) intoxication. Despite this overwhelming association between EtOH intoxication and injury, relatively little attention has been paid to determining the role of EtOH in post-injury pathogenesis. This article reviews studies which have evaluated the impact of EtOH on post-burn intestinal immunity and barrier functions. The findings from these studies suggest that while a smaller burn injury by itself may not have an adverse effect on host defense, when combined with prior EtOH intoxication it may become detrimental. Experimental data from our laboratory further supports the notion that EtOH intoxication before burn injury suppresses intestinal immune defense, impairs gut barrier functions, and increases bacterial growth. This results in increased bacterial translocation which may contribute to post injury pathogenesis. Altogether, the studies reviewed in this article suggest that EtOH intoxication at the time of injury is a risk factor, and therefore blood EtOH should be checked in burn/trauma patients at the time of hospital admission. PMID- 19561987 TI - Pediatric procedural sedation and analgesia. AB - Procedural sedation and analgesia (PSA) is an evolving field in pediatric emergency medicine. As new drugs breach the boundaries of anesthesia in the Pediatric Emergency Department, parents, patients, and physicians are finding new and more satisfactory methods of sedation. Short acting, rapid onset agents with little or no lingering effects and improved safety profiles are replacing archaic regimens. This article discusses the warning signs and areas of a patient's medical history that are particularly pertinent to procedural sedation and the drugs used. The necessary equipment is detailed to provide the groundwork for implementing safe sedation in children. It is important for practitioners to familiarize themselves with a select few of the PSA drugs, rather than the entire list of sedatives. Those agents most relevant to PSA in the pediatric emergency department are presented. PMID- 19561988 TI - Emergency treatment of a snake bite: Pearls from literature. AB - Snake bite is a well-known occupational hazard amongst farmers, plantation workers, and other outdoor workers and results in much morbidity and mortality throughout the world. This occupational hazard is no more an issue restricted to a particular part of the world; it has become a global issue. Accurate statistics of the incidence of snakebite and its morbidity and mortality throughout the world does not exist; however, it is certain to be higher than what is reported. This is because even today most of the victims initially approach traditional healers for treatment and many are not even registered in the hospital. Hence, registering such patients is an important goal if we are to have accurate statistics and reduce the morbidity and mortality due to snakebite. World Health Organization/South East Asian Region Organisation (WHO/SEARO) has published guidelines, specific for the South East Asian region, for the clinical management of snakebites. The same guidelines may be applied for managing snakebite patients in other parts of the world also, since no other professional body has come up with any other evidence-based guidelines. In this article we highlight the incidence and clinical features of different types of snakebite and the management guidelines as per the WHO/SEARO recommendation. PMID- 19561990 TI - Minor trauma triggering cervicofacial necrotizing fasciitis from odontogenic abscess. AB - Necrotizing fasciitis (NF) of the face and neck is a very rare complication of dental infection. Otolaryngologists and dentists should be familiar with this condition because of its similarity to odontogenic deep neck space infection in the initial stages, its rapid spread, and its life-threatening potential. Trauma has been reported to be an important predisposing factor for NF of the face. In this paper, we describe the presentation and treatment of a 62-year-old man who developed NF of the face and neck following bilateral odontogenic deep neck space abscesses. The disease progressed rapidly, with necrosis of the skin, after the patient inflicted minor trauma in the form of application of heated medicinal leaves. The organism isolated in culture from pus was Acinetobacter sp. The comorbid conditions in our patient were anemia and chronic alcoholism. The patient was managed by immediate and repeated extensive debridements and split skin grafting. PMID- 19561991 TI - Epstein-Barr virus-associated hemophagocytic syndrome mimicking severe sepsis. AB - Severe sepsis is amongst the most common reasons for admission to the intensive care unit (ICU) throughout the world and is a common cause of death. The diagnosis of sepsis is usually straightforward, being based on a constellation of clinical and laboratory features. Noninfectious disorders, including pancreatitis, drug reactions, and autoimmune disorders, may cause a systemic inflammatory response that mimics sepsis. We present the case of a 32-year-old male with Epstein-Barr virus-associated hemophagocytic syndrome who presented to the ICU with features of severe sepsis which progressed to multisystem organ failure and death despite aggressive supportive measures. PMID- 19561989 TI - Mucormycosis in immunochallenged patients. AB - Mucorales species are deadly opportunistic fungi with a rapidly invasive nature. A rare disease, mucormycosis is most commonly reported in patients with diabetes mellitus, because the favorable carbohydrate-rich environment allows the Mucorales fungi to flourish, especially in the setting of ketoacidosis. However, case reports over the past 20 years show that a growing number of cases of mucormycosis are occurring during treatment following bone marrow transplants (BMT) and hematological malignancies (HM) such as leukemia and lymphoma. This is due to the prolonged treatment of these patients with steroids and immunosuppressive agents. Liposomal amphotericin B treatment and posaconazole are two pharmacologic agents that seem to be effective against mucormycosis, but the inherently rapid onset and course of the disease, in conjunction with the difficulty in correctly identifying it, hinder prompt institution of appropriate antifungal therapy. This review of the literature discusses the clinical presentation, diagnosis, and treatment of mucormycosis among the BMT and HM populations. PMID- 19561992 TI - Complication following primary repair of a penetrating bull horn injury to the trachea. AB - A 22-year-old male patient was admitted to the casualty with a bull horn injury in the lower zone of the neck in the midline. The patient was conscious and distressed but hemodynamically stable. Local examination revealed a lacerated wound. He underwent emergency primary repair of the wound under halothane anesthesia; intubation was done keeping in readiness all preparations for difficult airway management. Postoperatively, elective controlled ventilation was performed with continuous infusion of muscle relaxant. After approximately 8 hours of controlled ventilation, the syringe pump failed; this initially went unnoticed and made the patient cough and buck on the tube. Infusion was restarted after a bolus dose of vecuronium bromide intravenously but, meanwhile, the patient developed subcutaneous emphysema in the neck. He was immediately transferred to the operating room, where exploration of the surgical site revealed dehiscence of the tracheal wound; this had led to the subcutaneous emphysema. Repair of the tracheal wound dehiscence was not possible due to both lack of space and lack of tissue for apposition. Hence, a tracheostomy tube was inserted through the tracheal wound and the patient was transferred to the intensive care unit for elective controlled ventilation. The patient was weaned off the ventilator within 24 h and transferred to the surgical ward on spontaneous ventilation with the tracheostomy tube in situ. The size of the patient's tracheostomy tube was reduced gradually by the serial exchange method. The wound ultimately healed with minimal scarring. PMID- 19561993 TI - The value of MR imaging in posttraumatic diffuse axonal injury. PMID- 19561994 TI - ECG J waves. PMID- 19561995 TI - Head injuries in a rural setup: Challenges and potential solutions. PMID- 19561997 TI - Whither CME. PMID- 19561998 TI - Measuring up to Medication Safety In Hospitals. PMID- 19561996 TI - The potential adverse patient effects of ambulance ramping, a relatively new problem at the interface between prehospital and ED care. PMID- 19561999 TI - Pharmacy product tracing likely to go national: costs to pharmacies worrisome. PMID- 19562000 TI - New drugs drug news. PMID- 19562001 TI - Economic instability and its impact on decision making in health care. PMID- 19562002 TI - Pharmaceutical approval update. PMID- 19562003 TI - American heart association 2008 scientific sessions United European gastroenterology week american college of rheumatology scientific meeting. PMID- 19562004 TI - Rosacea: a review. AB - Rosacea is a chronic inflammatory condition of the facial skin affecting the blood vessels and pilosebaceous units. Rosacea is more common in persons of northern and western European descent with a fair complexion, but it can affect skin of any color. Although symptoms may wax and wane during the short term, rosacea can progress with time. Patients usually present with complaints of flushing and blushing and sensitive skin, and their skin may be especially irritated by topical preparations. Rosacea has a variety of triggers; however, they may be unnoticed by the patient.Standard treatments approved by the FDA include azelaic acid, topical metronidazole, and oral tetracyclines, in particular minocycline and doxycycline. Other topical treatments include topical clindamycin, subantimicrobial-dose doxycycline, and sulfur products. Azithromycin and controlled-release minocycline are possible options for treating rosacea, but the FDA has not approved either agent for this indication. PMID- 19562006 TI - Enforcing medical licensing in Illinois: 1877-1890. PMID- 19562007 TI - Endless Forms: Charles Darwin, Natural Science and the Visual Arts. PMID- 19562005 TI - Use with caution: developmental systems divergence and potential pitfalls of animal models. AB - Transgenic animal models have played an important role in elucidating gene functions and the molecular basis development, physiology, behavior, and pathogenesis. Transgenic models have been so successful that they have become a standard tool in molecular genetics and biomedical studies and are being used to fulfill one of the main goals of the post-genomic era: to assign functions to each gene in the genome. However, the assumption that gene functions and genetic systems are conserved between models and humans is taken for granted, often in spite of evidence that gene functions and networks diverge during evolution. In this review, I discuss some mechanisms that generate functional divergence and highlight recent examples demonstrating that gene functions and regulatory networks diverge through time. These examples suggest that annotation of gene functions based solely on mutant phenotypes in animal models, as well as assumptions of conserved functions between species, can be wrong. Therefore, animal models of gene function and human disease may not provide appropriate information, particularly for rapidly evolving genes and systems. PMID- 19562008 TI - Experiences with obstetric fistula in rural Uganda. PMID- 19562009 TI - What happens when special needs children grow up? PMID- 19562011 TI - Asperger's syndrome in adulthood. AB - INTRODUCTION: Asperger's syndrome is one of the autism spectrum disorders. Affected individuals display considerably impaired capacity for social interaction, unusual special interests, and a tendency towards ritualized behavior. METHODS: The etiology, symptoms, diagnosis, and treatment of Asperger's syndrome in adulthood are outlined on the basis of a selective literature review via Medline and information in relevant reference books. Furthermore, the authors report their personal experience at a special clinic for adults. RESULTS: Asperger's syndrome in adulthood can be diagnosed by thorough anamnesis, heteroanamnesis-with emphasis on childhood-and painstaking clinical examination. The considerable psychosocial impairments affect the patients' professional, social, and private lives. The precise etiology is still unknown, but a multifactorial origin with genetic, neurobiological, and psychosocial components appears probable. Although no specific, empirically tested treatment concepts have yet been established, psychotherapeutic elements (structuring and directive interventions) seem to be helpful, together with pharmacotherapy-if indicated-in the presence of comorbidity. CONCLUSIONS: Asperger's syndrome should be included in the differential diagnosis of adults who display the corresponding symptoms. The etiopathogenesis and treatment of Asperger's syndrome in adulthood should be further investigated. PMID- 19562010 TI - What is stochastic resonance? Definitions, misconceptions, debates, and its relevance to biology. AB - Stochastic resonance is said to be observed when increases in levels of unpredictable fluctuations--e.g., random noise--cause an increase in a metric of the quality of signal transmission or detection performance, rather than a decrease. This counterintuitive effect relies on system nonlinearities and on some parameter ranges being "suboptimal". Stochastic resonance has been observed, quantified, and described in a plethora of physical and biological systems, including neurons. Being a topic of widespread multidisciplinary interest, the definition of stochastic resonance has evolved significantly over the last decade or so, leading to a number of debates, misunderstandings, and controversies. Perhaps the most important debate is whether the brain has evolved to utilize random noise in vivo, as part of the "neural code". Surprisingly, this debate has been for the most part ignored by neuroscientists, despite much indirect evidence of a positive role for noise in the brain. We explore some of the reasons for this and argue why it would be more surprising if the brain did not exploit randomness provided by noise--via stochastic resonance or otherwise--than if it did. We also challenge neuroscientists and biologists, both computational and experimental, to embrace a very broad definition of stochastic resonance in terms of signal-processing "noise benefits", and to devise experiments aimed at verifying that random variability can play a functional role in the brain, nervous system, or other areas of biology. PMID- 19562013 TI - Noise research is hampered. PMID- 19562012 TI - Cardiac arrest: composition of resuscitation teams and training techniques: results of a hospital survey in German-speaking countries. AB - BACKGROUND: The long-term outcome of patients requiring cardiopulmonary resuscitation depends heavily on swift and appropriate care. The aim of this study was to obtain data on the composition and training of resuscitation teams in specialist departments for internal medicine and anesthesiology. METHODS: Between October 2006 and February 2007, 440 questionnaires were sent to departments for anesthesiology and internal medicine in Germany (hospitals with more than 300 beds) and to university hospitals in Switzerland and Austria. RESULTS: The response rate was 38%. Of 166 participating hospitals, 152 have an emergency team. Resuscitation training (RT) takes place in 111 hospitals. Ninety two hospitals (55%) hold a course more than once a year. Of those hospitals with RT, 86% use a simulation dummy, 77% conduct theoretical tutorials, and 65% follow a fixed algorithm. CONCLUSION: The majority of hospitals that participated in this survey have an emergency team in place and organize resuscitation training for their medical personnel. The training varies greatly, however, in frequency, size of group, and qualification of the trainer. Implementation of standardized training for and management of in-hospital resuscitation measures might further hone staff skills and therefore improve the long-term outcome for the patients concerned. PMID- 19562015 TI - Lyme disease--current state of knowledge. AB - BACKGROUND: Lyme disease is the most frequent tick-borne infectious disease in Europe. The discovery of the causative pathogen Borrelia burgdorferi in 1982 opened the way for the firm diagnosis of diseases in several clinical disciplines and for causal antibiotic therapy. At the same time, speculation regarding links between Borrelia infection and a variety of nonspecific symptoms and disorders resulted in overdiagnosis and overtreatment of suspected Lyme disease. METHOD: The authors conducted a selective review of the literature, including various national and international guidelines. RESULTS: The spirochete Borrelia burgdorferi sensu lato is present in approximately 5% to 35% of sheep ticks (Ixodes ricinus) in Germany, depending on the region. In contrast to North America, different genospecies are found in Europe. The most frequent clinical manifestation of Borrelia infection is erythema migrans, followed by neuroborreliosis, arthritis, acrodermatitis chronica atrophicans, and lymphocytosis benigna cutis. Diagnosis is made on the basis of the clinical symptoms, and in stages II and III by detection of Borrelia-specific antibodies. In adults erythema migrans is treated with doxycycline, in children with amoxicillin. The standard treatment of neuroborreliosis is third-generation cephalosporins. CONCLUSIONS: After appropriate antibiotic therapy, the outcome is favorable. In approximately 95% of cases neuroborreliosis is cured without long term sequelae. When chronic borreliosis is suspected, other potential causes of the clinical syndrome must be painstakingly excluded. PMID- 19562017 TI - The assessment of environmental and occupational exposure to hazardous substances by biomonitoring. AB - BACKGROUND: Modern biomonitoring has expanded beyond its origins in occupational medicine to cover a wide variety of diagnostic procedures and assessments of environmental pollution, ranging from exposure to heavy metals and chemicals to the effects of pesticides and environmental tobacco smoke. In biomonitoring, the overall pollutant load and hazardous exposure of an organism is quantitatively determined, by monitoring the pollutants themselves, their metabolic products and/or conjugates with protein or DNA, in either serum, urine or other body fluids, as well as tissue samples in exceptional circumstances. METHODS: A selective survey of the current literature is exemplified by our recent scientific and clinical experience. RESULTS: Biomonitoring is an excellent way to monitor potentially hazardous substances, especially for the assessment of systemic uptake from chronic exposure and the evaluation of subsequent health risks. Investigative biomonitoring can highlight incidental/accidental intoxication in individuals and provide new categories of problematic pollutants relevant to the general population. CONCLUSIONS: In combination with diagnostic procedures, biomonitoring provides an important contribution to the rational assessment of currently recorded pollutants and resulting health risks. It is as an evaluation tool available in daily practice as well as in health and environmental research applications. Legislation is already implemented that incorporates biomonitoring within the remit of specified occupational health screening. PMID- 19562016 TI - Obstetric brachial plexus palsy: treatment strategy, long-term results, and prognosis. AB - BACKGROUND: Obstetric brachial plexus palsy is rare, but the limb impairments are manifold and often long-lasting. Physiotherapy, microsurgical nerve reconstruction, secondary joint corrections, and muscle transpositions are employed with success. The role of conservative and operative treatment options should be regularly reviewed. METHODS: Selective literature review (evidence levels 3 and 4) and analysis of personal clinical operative and scientific experience over the past 15 years. RESULTS: Children with upper and total plexus palsy displaying nerve root avulsions and/or -ruptures are treated today by early primary nerve reconstruction in the first few months of life followed by secondary corrections, with good functional results. The late complications, with muscle weakness, impaired motion patterns, and joint dysplasia, are often underrated. CONCLUSIONS: The potential for scientific analysis is limited, due to the rarity and interindividual variability of the lesions and the varying effects on function and growth. Expectations and compliance are different in every patient. Surgical techniques are not yet standardized. Knowledge of the consequences for joint growth and congruence is inadequate. Today, functional improvement can be achieved by surgery in most clinical manifestations of obstetric brachial plexus palsy, within the framework of an interdisciplinary treatment concept. PMID- 19562018 TI - Re: Allogeneic stem cell transplantation in acute myeloid leukemia--establishment of indications on the basis of individual risk stratification. Statement was not sufficiently supported. PMID- 19562021 TI - Critical appraisal of scientific articles: part 1 of a series on evaluation of scientific publications. AB - INTRODUCTION: In the era of evidence-based medicine, one of the most important skills a physician needs is the ability to analyze scientific literature critically. This is necessary to keep medical knowledge up to date and to ensure optimal patient care. The aim of this paper is to present an accessible introduction into critical appraisal of scientific articles. METHODS: Using a selection of international literature, the reader is introduced to the principles of critical reading of scientific articles in medicine. For the sake of conciseness, detailed description of statistical methods is omitted. RESULTS: Widely accepted principles for critically appraising scientific articles are outlined. Basic knowledge of study design, structuring of an article, the role of different sections, of statistical presentations as well as sources of error and limitation are presented. The reader does not require extensive methodological knowledge. As far as necessary for critical appraisal of scientific articles, differences in research areas like epidemiology, clinical, and basic research are outlined. Further useful references are presented. CONCLUSION: Basic methodological knowledge is required to select and interpret scientific articles correctly. PMID- 19562022 TI - Radiosurgery for spinal malignant tumors. AB - BACKGROUND: Radiosurgery is a special treatment method that employs highly focused radiation to destroy tumors with high precision in a single session. A broad base of scientific evidence already exists for the radiosurgical treatment of brain metastases. Recent advances in medical technology now allow radiosurgery to be extended to the spine as well. METHODS: Selective literature review based on a PubMed search using the search terms stereotaxis, radiosurgery, stereotactic radiotherapy, accuracy, quality assurance, spine, spine metastasis, pain, Novalis, CyberKnife, Synergy, and robotics. We also present and analyze our own data as an illustration of the application of spinal radiosurgery. RESULTS: The literature search identified 20 scientific original publications and one recent review. The data indicate that, within the specific constraints of the method, radiosurgery can arrest the growth of up to 96% of spinal metastases. Durable pain relief can be achieved in patients with tumor-associated pain syndromes. The morbidity of spinal radiosurgery is low, with a less than 1% risk of myelopathy. CONCLUSIONS: Spinal radiosurgery is an independent, essentially noninvasive method of treatment. Different types of radiosurgical treatment apparatus are available. For properly selected patients, radiosurgery offers a good chance of therapeutic success with relatively rare complications. PMID- 19562023 TI - Different Products. PMID- 19562024 TI - Cardiovascular side effects. PMID- 19562027 TI - Stringent elimination diet. PMID- 19562029 TI - Growth of the human corpus callosum: modular and laminar morphogenetic zones. AB - The purpose of this focused review is to present and discuss recent data on the changing organization of cerebral midline structures that support the growth and development of the largest commissure in humans, the corpus callosum. We will put an emphasis on the callosal growth during the period between 20 and 45 postconceptual weeks (PCW) and focus on the advantages of a correlated histological/magnetic resonance imaging (MRI) approach. The midline structures that mediate development of the corpus callosum in rodents, also mediate its early growth in humans. However, later phases of callosal growth in humans show additional medial transient structures: grooves made up of callosal septa and the subcallosal zone. These modular (septa) and laminar (subcallosal zone) structures enable the growth of axons along the ventral callosal tier after 18 PCW, during the rapid increase in size of the callosal midsagittal cross-section area. Glial fibrillary acidic protein positive cells, neurons, guidance molecule semaphorin3A in cells and extracellular matrix (ECM), and chondroitin sulfate proteoglycan in the ECM have been identified along the ventral callosal tier in the protruding septa and subcallosal zone. Postmortem MRI at 3 T can demonstrate transient structures based on higher water content in ECM, and give us the possibility to follow the growth of the corpus callosum in vivo, due to the characteristic MR signal. Knowledge about structural properties of midline morphogenetic structures may facilitate analysis of the development of interhemispheric connections in the normal and abnormal fetal human brain. PMID- 19562030 TI - A novel in vitro multiple-stress dormancy model for Mycobacterium tuberculosis generates a lipid-loaded, drug-tolerant, dormant pathogen. AB - BACKGROUND: Mycobacterium tuberculosis (Mtb) becomes dormant and phenotypically drug resistant when it encounters multiple stresses within the host. Inability of currently available drugs to kill latent Mtb is a major impediment to curing and possibly eradicating tuberculosis (TB). Most in vitro dormancy models, using single stress factors, fail to generate a truly dormant Mtb population. An in vitro model that generates truly dormant Mtb cells is needed to elucidate the metabolic requirements that allow Mtb to successfully go through dormancy, identify new drug targets, and to screen drug candidates to discover novel drugs that can kill dormant pathogen. METHODOLOGY/PRINCIPAL FINDINGS: We developed a novel in vitro multiple-stress dormancy model for Mtb by applying combined stresses of low oxygen (5%), high CO(2) (10%), low nutrient (10% Dubos medium) and acidic pH (5.0), conditions Mtb is thought to encounter in the host. Under this condition, Mtb stopped replicating, lost acid-fastness, accumulated triacylglycerol (TG) and wax ester (WE), and concomitantly acquired phenotypic antibiotic-resistance. Putative neutral lipid biosynthetic genes were up regulated. These genes may serve as potential targets for new antilatency drugs. The triacylglycerol synthase1 (tgs1) deletion mutant, with impaired ability to accumulate TG, exhibited a lesser degree of antibiotic tolerance and complementation restored antibiotic tolerance. Transcriptome analysis with microarray revealed the achievement of dormant state showing repression of energy generation, transcription and translation machineries and induction of stress responsive genes. We adapted this model for drug screening using the Alamar Blue dye to quantify the antibiotic tolerant dormant cells. CONCLUSIONS/SIGNIFICANCE: The new in vitro multiple stress dormancy model efficiently generates Mtb cells meeting all criteria of dormancy, and this method is adaptable to high-throughput screening for drugs that can kill dormant Mtb. A critical link between storage lipid accumulation and development of phenotypic drug-resistance in Mtb was established. Storage lipid biosynthetic genes may be appropriate targets for novel drugs that can kill latent Mtb. PMID- 19562031 TI - Low-abundance HIV drug-resistant viral variants in treatment-experienced persons correlate with historical antiretroviral use. AB - BACKGROUND: It is largely unknown how frequently low-abundance HIV drug-resistant variants at levels under limit of detection of conventional genotyping (<20% of quasi-species) are present in antiretroviral-experienced persons experiencing virologic failure. Further, the clinical implications of low-abundance drug resistant variants at time of virologic failure are unknown. METHODOLOGY/PRINCIPAL FINDINGS: Plasma samples from 22 antiretroviral-experienced subjects collected at time of virologic failure (viral load 1380 to 304,000 copies/mL) were obtained from a specimen bank (from 2004-2007). The prevalence and profile of drug-resistant mutations were determined using Sanger sequencing and ultra-deep pyrosequencing. Genotypes were interpreted using Stanford HIV database algorithm. Antiretroviral treatment histories were obtained by chart review and correlated with drug-resistant mutations. Low-abundance drug-resistant mutations were detected in all 22 subjects by deep sequencing and only in 3 subjects by Sanger sequencing. In total they accounted for 90 of 247 mutations (36%) detected by deep sequencing; the majority of these (95%) were not detected by standard genotyping. A mean of 4 additional mutations per subject were detected by deep sequencing (p<0.0001, 95%CI: 2.85-5.53). The additional low abundance drug-resistant mutations increased a subject's genotypic resistance to one or more antiretrovirals in 17 of 22 subjects (77%). When correlated with subjects' antiretroviral treatment histories, the additional low-abundance drug resistant mutations correlated with the failing antiretroviral drugs in 21% subjects and correlated with historical antiretroviral use in 79% subjects (OR, 13.73; 95% CI, 2.5-74.3, p = 0.0016). CONCLUSIONS/SIGNIFICANCE: Low-abundance HIV drug-resistant mutations in antiretroviral-experienced subjects at time of virologic failure can increase a subject's overall burden of resistance, yet commonly go unrecognized by conventional genotyping. The majority of unrecognized resistant mutations correlate with historical antiretroviral use. Ultra-deep sequencing can provide important historical resistance information for clinicians when planning subsequent antiretroviral regimens for highly treatment-experienced patients, particularly when their prior treatment histories and longitudinal genotypes are not available. PMID- 19562032 TI - Rapid assessment of malaria transmission using age-specific sero-conversion rates. AB - BACKGROUND: Malaria transmission intensity is a crucial determinant of malarial disease burden and its measurement can help to define health priorities. Rapid, local estimates of transmission are required to focus resources better but current entomological and parasitological methods for estimating transmission intensity are limited in this respect. An alternative is determination of antimalarial antibody age-specific sero-prevalence to estimate sero-conversion rates (SCR), which have been shown to correlate with transmission intensity. This study evaluated SCR generated from samples collected from health facility attendees as a tool for a rapid assessment of malaria transmission intensity. METHODOLOGY AND PRINCIPAL FINDINGS: The study was conducted in north east Tanzania. Antibodies to Plasmodium falciparum merozoite antigens MSP-1(19) and AMA-1 were measured by indirect ELISA. Age-specific antibody prevalence was analysed using a catalytic conversion model based on maximum likelihood to generate SCR. A pilot study, conducted near Moshi, found SCRs for AMA-1 were highly comparable between samples collected from individuals in a conventional cross-sectional survey and those collected from attendees at a local health facility. For the main study, 3885 individuals attending village health facilities in Korogwe and Same districts were recruited. Both malaria parasite prevalence and sero-positivity were higher in Korogwe than in Same. MSP-1(19) and AMA-1 SCR rates for Korogwe villages ranged from 0.03 to 0.06 and 0.07 to 0.21 respectively. In Same district there was evidence of a recent reduction in transmission, with SCR among those born since 1998 [MSP-1(19) 0.002 to 0.008 and AMA-1 0.005 to 0.014 ] being 5 to 10 fold lower than among individuals born prior to 1998 [MSP-1(19) 0.02 to 0.04 and AMA-1 0.04 to 0.13]. Current health facility specific estimates of SCR showed good correlations with malaria incidence rates in infants in a contemporaneous clinical trial (MSP-1(19) r(2) = 0.78, p<0.01 & AMA-1 r(2) = 0.91, p<0.001). CONCLUSIONS: SCRs generated from age-specific anti malarial antibody prevalence data collected via health facility surveys were robust and credible. Analysis of SCR allowed detection of a recent drop in malaria transmission in line with recent data from other areas in the region. This health facility-based approach represents a potential tool for rapid assessment of recent trends in malaria transmission intensity, generating valuable data for local and national malaria control programs to target, monitor and evaluate their control strategies. PMID- 19562033 TI - Temporal and spatial analysis of clinical and molecular parameters in dextran sodium sulfate induced colitis. AB - BACKGROUND: Inflammatory bowel diseases (IBD), including mainly ulcerative colitis (UC) and Crohn's disease (CD), are inflammatory disorders of the gastrointestinal tract caused by an interplay of genetic and environmental factors. Murine colitis model induced by Dextran Sulfate Sodium (DSS) is an animal model of IBD that is commonly used to address the pathogenesis of IBD as well as to test efficacy of therapies. In this study we systematically analyzed clinical parameters, histological changes, intestinal barrier properties and cytokine profile during the colitic and recovery phase. METHODS: C57BL/6 mice were administered with 3.5% of DSS in drinking water for various times. Clinical and histological features were determined using standard criteria. Myeloperoxidase (MPO) activity, transepithelial permeability and proinflammatory mediators were determined in whole colon or proximal and distal parts of colon. RESULTS: As expected after administration of DSS, mice manifest loss of body weight, shortening of colon length and bloody feces. Histological manifestations included shortening and loss of crypts, infiltration of lymphocytes and neutrophil, symptoms attenuated after DSS withdrawal. The MPO value, as inflammation indicator, also increases significantly at all periods of DSS treatment, and even after DSS withdrawal, it still held at very high levels. Trans-mucosal permeability increased during DSS treatment, but recovered to almost control level after DSS withdrawal. The production of proinflammatory mediators by colonic mucosa were enhanced during DSS treatment, and then recovered to pre-treated level after DSS withdrawal. Finally, enhanced expression of proinflammatory mediators also revealed a different profile feature in proximal and distal parts of the colon. CONCLUSION: Experimental colitis induced by DSS is a good animal model to study the mechanisms underlying the pathogenesis and intervention against IBD, especially UC. PMID- 19562034 TI - A genetic screen in Drosophila reveals novel cytoprotective functions of the autophagy-lysosome pathway. AB - The highly conserved autophagy-lysosome pathway is the primary mechanism for breakdown and recycling of macromolecular and organellar cargo in the eukaryotic cell. Autophagy has recently been implicated in protection against cancer, neurodegeneration, and infection, and interest is increasing in additional roles of autophagy in human health, disease, and aging. To search for novel cytoprotective features of this pathway, we carried out a genetic mosaic screen for mutations causing increased lysosomal and/or autophagic activity in the Drosophila melanogaster larval fat body. By combining Drosophila genetics with live-cell imaging of the fluorescent dye LysoTracker Red and fixed-cell imaging of autophagy-specific fluorescent protein markers, the screen was designed to identify essential metazoan genes whose disruption causes increased flux through the autophagy-lysosome pathway. The screen identified a large number of genes associated with the protein synthesis and ER-secretory pathways (e.g. aminoacyl tRNA synthetases, Oligosaccharyl transferase, Sec61alpha), and with mitochondrial function and dynamics (e.g. Rieske iron-sulfur protein, Dynamin-related protein 1). We also observed that increased lysosomal and autophagic activity were consistently associated with decreased cell size. Our work demonstrates that disruption of the synthesis, transport, folding, or glycosylation of ER-targeted proteins at any of multiple steps leads to autophagy induction. In addition to illuminating cytoprotective features of autophagy in response to cellular damage, this screen establishes a genetic methodology for investigating cell biological phenotypes in live cells, in the context of viable wild type organisms. PMID- 19562035 TI - Nestin modulates glucocorticoid receptor function by cytoplasmic anchoring. AB - Nestin is the characteristic intermediate filament (IF) protein of rapidly proliferating progenitor cells and regenerating tissue. Nestin copolymerizes with class III IF-proteins, mostly vimentin, into heteromeric filaments. Its expression is downregulated with differentiation. Here we show that a strong nestin expression in mouse embryo tissue coincides with a strong accumulation of the glucocorticoid receptor (GR), a key regulator of growth and differentiation in embryonic development. Microscopic studies on cultured cells show an association of GR with IFs composed of vimentin and nestin. Cells lacking nestin, but expressing vimentin, or cells expressing vimentin, but lacking nestin accumulate GR in the nucleus. Completing these networks with an exogenous nestin, respectively an exogenous vimentin restores cytoplasmic anchoring of GR to the IF system. Thus, heteromeric filaments provide the basis for anchoring of GR. The reaction pattern with phospho-GR specific antibodies and the presence of the chaperone HSC70 suggest that specifically the unliganded receptor is anchored to the IF system. Ligand addition releases GR from IFs and shifts the receptor into the nucleus. Suppression of nestin by specific shRNA abolishes anchoring of GR, induces its accumulation in the nucleus and provokes an irreversible G1/S cell cycle arrest. Suppression of GR prior to that of nestin prevents entry into the arrest. The data give evidence that nestin/vimentin specific anchoring modulates growth suppression by GR. We hypothesize that expression of nestin is a major determinant in suppression of anti-proliferative activity of GR in undifferentiated tissue and facilitates activation of this growth control in a precise tissue and differentiation dependent manner. PMID- 19562036 TI - Rosiglitazone and myocardial infarction in patients previously prescribed metformin. AB - OBJECTIVE: Rosiglitazone was found associated with approximately a 43% increase in risk of acute myocardial infarction (AMI) in a two meta-analyses of clinical trials. Our objective is to estimate the magnitude of the association in real world patients previously treated with metformin. RESEARCH DESIGN AND METHODS: We conducted a nested case control study in British Columbia using health care databases on 4.3 million people. Our cohort consisted of 158,578 patients with Type 2 diabetes who used metformin as first-line drug treatment. We matched 2,244 cases of myocardial infarction (AMI) with up to 4 controls. Conditional logistic regression models were used to estimate matched odds ratios for AMI associated with treatment with rosiglitazone, pioglitazone and sulfonylureas. RESULTS: In our cohort of prior metformin users, adding rosiglitazone for up to 6 months was not associated with an increased risk of AMI compared to adding a sulfonylurea (odds ratio [OR] 1.38; 95% confidence interval [CI], 0.91-2.10), or compared to adding pioglitazone (OR for rosi versus pio 1.41; 95% CI, 0.74-2.66). There were also no significant differences between rosiglitazone, pioglitazone and sulfonylureas for longer durations of treatment. Though not significantly different from sulfonylureas, there was a transient increase in AMI risk associated with the first 6 months of treatment with a glitazone compared to not using the treatment (OR 1.53; 95% CI, 1.13-2.07) CONCLUSIONS: In our British Columbia cohort of patients who received metformin as first-line pharmacotherapy for Type 2 diabetes mellitus, further treatment with rosiglitazone did not increase the risk of AMI compared to patients who were treated with pioglitazone or a sulfonylurea. Though not statistically significantly different compared from each other, an increased risk of AMI observed after starting rosiglitazone or sulfonylureas is a matter of concern that requires more research. PMID- 19562037 TI - A comprehensive approach to identification of surface-exposed, outer membrane spanning proteins of Leptospira interrogans. AB - Leptospirosis is a zoonosis with worldwide distribution caused by pathogenic spirochetes belonging to the genus Leptospira. The leptospiral life cycle involves transmission via fresh water and colonization of the renal tubules of their reservoir hosts or infection of accidental hosts, including humans. Bacterial outer membrane proteins (OMPs), particularly those with surface-exposed regions, play crucial roles in virulence mechanisms of pathogens and the adaptation to various environmental conditions, including those of the mammalian host. Little is known about the surface-exposed OMPs in Leptospira, particularly those with outer membrane-spanning domains. Herein, we describe a comprehensive strategy for identification and characterization of leptospiral transmembrane OMPs. The genomic sequence of L. interrogans serovar Copenhageni strain Fiocruz L1-130 allowed us to employ the beta-barrel prediction programs, PRED-TMBB and TMBETA-NET, to identify potential transmembrane OMPs. Several complementary methods were used to characterize four novel OMPs, designated OmpL36, OmpL37, OmpL47 and OmpL54. In addition to surface immunofluorescence and surface biotinylation, we describe surface proteolysis of intact leptospires as an improved method for determining the surface exposure of leptospiral proteins. Membrane integration was confirmed using techniques for removal of peripheral membrane proteins. We also demonstrate deficiencies in the Triton X-114 fractionation method for assessing the outer membrane localization of transmembrane OMPs. Our results establish a broadly applicable strategy for the elucidation of novel surface-exposed outer membrane-spanning proteins of Leptospira, an essential step in the discovery of potential virulence factors, diagnostic antigens and vaccine candidates. PMID- 19562040 TI - On Linear Infeasibility Arising in Intensity-Modulated Radiation Therapy Inverse Planning. AB - Intensity-modulated radiation therapy (IMRT) gives rise to systems of linear inequalities, representing the effects of radiation on the irradiated body. These systems are often infeasible, in which case one settles for an approximate solution, such as an {alpha, beta}-relaxation, meaning that no more than alpha percent of the inequalities are violated by no more than beta percent. For real world IMRT problems, there is a feasible {alpha, beta}-relaxation for sufficiently large alpha, beta > 0, however large values of these parameters may be unacceptable medically.The {alpha, beta}-relaxation problem is combinatorial, and for given values of the parameters can be solved exactly by Mixed Integer Programming (MIP), but this may be impractical because of problem size, and the need for repeated solutions as the treatment progresses.As a practical alternative to the MIP approach we present a heuristic non-combinatorial method for finding an approximate relaxation. The method solves a Linear Program (LP) for each pair of values of the parameters {alpha, beta} and progresses through successively increasing values until an acceptable solution is found, or is determined non-existent. The method is fast and reliable, since it consists of solving a sequence of LP's. PMID- 19562041 TI - A new method for eliciting three speaking styles in the laboratory. AB - In this study, a method was developed to elicit three different speaking styles, reduced, citation, and hyperarticulated, using controlled sentence materials in a laboratory setting. In the first set of experiments, the reduced style was elicited by having twelve talkers read a sentence while carrying out a distractor task that involved recalling from short-term memory an individually-calibrated number of digits. The citation style corresponded to read speech in the laboratory. The hyperarticulated style was elicited by prompting talkers (twice) to reread the sentences more carefully. The results of perceptual tests with naive listeners and an acoustic analysis showed that six of the twelve talkers produced a reduced style of speech for the test sentences in the distractor task relative to the same sentences in the citation style condition. In addition, all talkers consistently produced sentences in the citation and hyperarticulated styles. In the second set of experiments, the reduced style was elicited by increasing the number of digits in the distractor task by one (a heavier cognitive load). The procedures for eliciting citation and hyperarticulated sentences remained unchanged. Ten talkers were recorded in the second experiment. The results showed that six out of ten talkers differentiated all three styles as predicted (70% of all sentences recorded). In addition, all talkers consistently produced sentences in the citation and hyperarticulated styles. Overall, the results demonstrate that it is possible to elicit controlled sentence stimulus materials varying in speaking style in a laboratory setting, although the method requires further refinement to elicit these styles more consistently from individual participants. PMID- 19562038 TI - Peroxiredoxin 6 fails to limit phospholipid peroxidation in lung from Cftr knockout mice subjected to oxidative challenge. AB - Oxidative stress plays a prominent role in the pathophysiology of cystic fibrosis (CF). Despite the presence of oxidative stress markers and a decreased antioxidant capacity in CF airway lining fluid, few studies have focused on the oxidant/antioxidant balance in CF cells. The aim of the current study was to investigate the cellular levels of reactive oxygen species (ROS), oxidative damage and enzymatic antioxidant defenses in the lung of Cftr-knockout mice in basal conditions and as a response to oxidative insult.The results show that endogenous ROS and lipid peroxidation levels are higher in Cftr(-/-) lung when compared to wild-type (Cftr(+/+)) in basal conditions, despite a strong enzymatic antioxidant response involving superoxide dismutases, glutathione peroxidases and peroxiredoxin 6 (Prdx6). The latter has the unique capacity to directly reduce membrane phospholipid hydroperoxides (PL-OOH). A dramatic increase in PL-OOH levels in Cftr(-/-) lung consecutive to in vivo oxidative challenge by paraquat (PQ) unmasks a susceptibility to phospholipid peroxidation. PQ strongly decreases Prdx6 expression in Cftr(-/-) mice compared to Cftr(+/+). Similar results were obtained after P. aeruginosa LPS challenge. Two-dimensional gel analysis of Prdx6 revealed one main molecular form in basal conditions and a PQ-induced form only detected in Cftr(+/+) lung. Mass spectrometry experiments suggested that, as opposed to the main basal form, the one induced by PQ is devoid of overoxidized catalytic Cys47 and could correspond to a fully active form that is not induced in Cftr(-/-) lung. These results highlight a constitutive redox imbalance and a vulnerability to oxidative insult in Cftr(-/-) lung and present Prdx6 as a key component in CF antioxidant failure. This impaired PL-OOH detoxification mechanism may enhance oxidative damage and stress-related signaling, contributing to an exaggerated inflammatory response in CF lung. PMID- 19562039 TI - Targeting 160 candidate genes for blood pressure regulation with a genome-wide genotyping array. AB - The outcome of Genome-Wide Association Studies (GWAS) has challenged the field of blood pressure (BP) genetics as previous candidate genes have not been among the top loci in these scans. We used Affymetrix 500K genotyping data of KORA S3 cohort (n = 1,644; Southern-Germany) to address (i) SNP coverage in 160 BP candidate genes; (ii) the evidence for associations with BP traits in genome-wide and replication data, and haplotype analysis. In total, 160 gene regions (genic region+/-10 kb) covered 2,411 SNPs across 11.4 Mb. Marker densities in genes varied from 0 (n = 11) to 0.6 SNPs/kb. On average 52.5% of the HAPMAP SNPs per gene were captured. No evidence for association with BP was obtained for 1,449 tested SNPs. Considerable associations (P<10(-3)) were detected for the genes, where >50% of HAPMAP SNPs were tagged. In general, genes with higher marker density (>0.2 SNPs/kb) revealed a better chance to reach close to significance associations. Although, none of the detected P-values remained significant after Bonferroni correction (P<0.05/2319, P<2.15 x 10(-5)), the strength of some detected associations was close to this level: rs10889553 (LEPR) and systolic BP (SBP) (P = 4.5 x 10(-5)) as well as rs10954174 (LEP) and diastolic BP (DBP) (P = 5.20 x 10(-5)). In total, 12 markers in 7 genes (ADRA2A, LEP, LEPR, PTGER3, SLC2A1, SLC4A2, SLC8A1) revealed considerable association (P<10(-3)) either with SBP, DBP, and/or hypertension (HYP). None of these were confirmed in replication samples (KORA S4, HYPEST, BRIGHT). However, supportive evidence for the association of rs10889553 (LEPR) and rs11195419 (ADRA2A) with BP was obtained in meta-analysis across samples stratified either by body mass index, smoking or alcohol consumption. Haplotype analysis highlighted LEPR and PTGER3. In conclusion, the lack of associations in BP candidate genes may be attributed to inadequate marker coverage on the genome-wide arrays, small phenotypic effects of the loci and/or complex interaction with life-style and metabolic parameters. PMID- 19562042 TI - Convenient and Efficient Synthesis of a Lanthanide-Coordinated, Diethylene Triamine Pentaacetic Acid Labeled Biopolymer as an Assay for the Cholecystokinin B Receptor. AB - To develop an assay for the cholecystokinin B receptor with an Eu(3+)-labeled cholecystokinin peptide via a diethylene triamine pentaacetic acid chelating linker, a commercial dianhydride diethylene triamine pentaacetic acid precursor was directly attached to the N-terminus of cholecystokinin peptides by a solid phase synthesis method with a satisfactory yield and purity after reverse-phase high-performance liquid chromatography separation. Lanthanide was then coordinated to the peptide via a diethylene triamine pentaacetic acid bifunctional agent. This method is a useful approach to the large-scale synthesis of lanthanide(3+)-coordinated, diethylene triamine pentaacetic acid labeled biopolymers. This research provides not only a simple and convenient method for the preparation of lanthanide-based peptide ligand libraries but also possible lanthanide-based high-throughput screening of peptide receptors with a timeresolved fluorescence assay system. Five biopolymers were synthesized and characterized with high-resolution electrospray ionization in this study. PMID- 19562043 TI - A Review of Challenges in the Use of fMRI for Disease Classification / Characterization and A Projection Pursuit Application from Multi-site fMRI Schizophrenia Study. AB - Functional magnetic resonance imaging (fMRI) is a fairly new technique that has the potential to characterize and classify brain disorders such as schizophrenia. It has the possibility of playing a crucial role in designing objective prognostic/diagnostic tools, but also presents numerous challenges to analysis and interpretation. Classification provides results for individual subjects, rather than results related to group differences. This is a more complicated endeavor that must be approached more carefully and efficient methods should be developed to draw generalized and valid conclusions out of high dimensional data with a limited number of subjects, especially for heterogeneous disorders whose pathophysiology is unknown. Numerous research efforts have been reported in the field using fMRI activation of schizophrenia patients and healthy controls. However, the results are usually not generalizable to larger data sets and require careful definition of the techniques used both in designing algorithms and reporting prediction accuracies. In this review paper, we survey a number of previous reports and also identify possible biases (cross-validation, class size, e.g.) in class comparison/prediction problems. Some suggestions to improve the effectiveness of the presentation of the prediction accuracy results are provided. We also present our own results using a projection pursuit algorithm followed by an application of independent component analysis proposed in an earlier study. We classify schizophrenia versus healthy controls using fMRI data of 155 subjects from two sites obtained during three different tasks. The results are compared in order to investigate the effectiveness of each task and differences between patients with schizophrenia and healthy controls were investigated. PMID- 19562045 TI - From the Editor's desk. PMID- 19562044 TI - Random Effects Models in a Meta-Analysis of the Accuracy of Two Diagnostic Tests Without a Gold Standard. AB - In studies of the accuracy of diagnostic tests, it is common that both the diagnostic test itself and the reference test are imperfect. This is the case for the microsatellite instability test, which is routinely used as a prescreening procedure to identify individuals with Lynch syndrome, the most common hereditary colorectal cancer syndrome. The microsatellite instability test is known to have imperfect sensitivity and specificity. Meanwhile, the reference test, mutation analysis, is also imperfect. We evaluate this test via a random effects meta analysis of 17 studies. Study-specific random effects account for between-study heterogeneity in mutation prevalence, test sensitivities and specificities under a nonlinear mixed effects model and a Bayesian hierarchical model. Using model selection techniques, we explore a range of random effects models to identify a best-fitting model. We also evaluate sensitivity to the conditional independence assumption between the microsatellite instability test and the mutation analysis by allowing for correlation between them. Finally, we use simulations to illustrate the importance of including appropriate random effects and the impact of overfitting, underfitting, and misfitting on model performance. Our approach can be used to estimate the accuracy of two imperfect diagnostic tests from a meta-analysis of multiple studies or a multicenter study when the prevalence of disease, test sensitivities and/or specificities may be heterogeneous among studies or centers. PMID- 19562047 TI - Recognising, understanding and managing endometriosis. AB - Endometriosis is defined as the presence of tissue lesions or nodules, histologically similar to the endometrium, at sites outside the uterus. It is a highly variable condition that has a wide spectrum of symptoms. The aetiology of endometriosis is probably multifactorial, with a strong familial component recognised. Women with endometriosis have multiple disturbances of function in the eutopic endometrium that women without the disease do not have. A firm diagnosis of endometriosis is rarely possible in general practice. The 'gold standard' for the diagnosis of pelvic endometriosis is currently a diagnostic laparoscopy. PMID- 19562046 TI - Ovarian tissue cryopreservation: An update. AB - Ovarian tissue cryopreservation and transplantation have been considered as promising means of fertility preservation for women who have survived cancer, with livebirths being reported from this technique. Ovarian tissue cryopreservation can be offered to patients with different types of cancer. Among the cryoprotectants, glycerol appears to give the poorest results. The techniques of cryopreserving ovarian tissue and alternative approaches have been reviewed in this article. The readers are reminded that this technique is still experimental and informed consent to be obtained from patients after counseling with medical information on the risks involved. PMID- 19562048 TI - Polycystic ovaries and infertility: Our experience. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is one of the most common (15-20%) endocrine disorders in women of childbearing age. Although it is a major cause of infertility, its etiology remains unknown and its treatment difficult. AIM: To evaluate the incidence, treatment and outcome of patients with PCOS. DESIGN: Retrospective analysis. MATERIALS AND METHODS: PCOS patients (914 of the 1057) attending the outpatient department (OPD) from June 2003 to February 2008 were evaluated for this study. Of the 914 patients investigated, 814 came for treatment and these patients were studied for hormonal disturbances and their response to various modalities of treatment. RESULTS: Of the 2270 infertility patients, 46.50% (1057) had PCOS, out of these, 86.47% (914) were investigated and 77% (814) came for treatment. Our overall pregnancy rate was 48.40% (394/814). The pregnancy rate per cycle with timed intercourse (TI) was 44.77% (47/105), 17.09% (286/1673) with intrauterine insemination (IUI), 29.82% (51/171) with in vitro fertilization (IVF) and 22.22% (10/45) with frozen embryo transfer (FET). The maximum number of pregnancies (85.29%, 284/333) were achieved in the first three treatment cycles. The abortion rate was 19.01% (73/384) and the incidence of ectopic pregnancy was 5.47% (21/384). Complications seen were in the form of ovarian hyperstimulation (OHSS), retention cyst on day two and multiple pregnancies in 11.71% (228/1946) of the total treatment cycles. CONCLUSION: Most PCOS symptoms could be adequately controlled or eliminated with proper diagnosis and treatment. Thus, ovulation induction (OI) protocols and treatment modalities must be balanced for optimal results. PMID- 19562049 TI - The outcome of ART in males with impaired spermatogenesis. AB - AIMS: This study was conducted to evaluate the outcomes of assisted reproductive technology (ART) procedures, viz., intrauterine insemination (IUI), in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in males with impaired spermatogenesis. SETTINGS AND DESIGN: The subjects of the study were infertile couples who were undergoing ART treatment due to male factor indications. The project was designed to correlate the outcome of the ART treatment and its efficacy in different study groups. METHODS: MALES WERE GROUPED AS: 1. oligozoospermia (n = 153), 2. asthenoteratozoospermia (n = 158), 3. obstructive azoospermia (n = 110) and 4. nonobstructive azoospermia (n = 58). Patients from groups 1 and 2 were considered for IUI, IVF and ICSI. Those from group 3 were considered for IVF and ICSI and the 4th group underwent only ICSI treatment. RESULTS: Oligozoospermia showed lower pregnancy rates with IUI than with both IVF and ICSI. An average minimum native and postharvest count was obtained to get an acceptable IUI outcome. Asthenoteratozoospermia had the lowest pregnancy rate with IUI as compared to IVF, whereas ICSI showed significantly higher pregnancy rates in this group. Obstructive azoospermia showed significant improvement with ICSI over IVF. In nonobstructive azoospermia, ICSI resulted in a 27.58% pregnancy rate. CONCLUSION: The IUI outcome was impressive though less effective whereas there was no difference between the IVF and ICSI outcomes in oligozoospermia. In asthenoteratozoospermia, ICSI showed a significant advantage over IUI and IVF, with IUI resulting in poor outcome in this group. In obstructive azoospermia, ICSI had a distinct advantage over IVF whereas in nonobstructive azoospermia, ICSI, the only option, was found to be effective and helpful in achieving an acceptable pregnancy rate. PMID- 19562050 TI - Changes in serum calcium, magnesium and inorganic phosphorus levels during different phases of the menstrual cycle. AB - BACKGROUND: The menstrual cycle is a complex process involving the interaction of the hypothalamus, the anterior pituitary, the ovaries and the uterus. The hormonal changes occurring during this cyclic process not only affect oocyte maturation and the endometrial and vaginal environments but can also have an effect on a number of other physiological and biochemical phenomena. AIM AND METHOD: We investigated the changes in serum calcium, magnesium and inorganic phosphorus levels during different phases of the menstrual cycle in fifty healthy young women. We found subtle but significant variations in these levels in the menstrual, follicular and luteal phases. RESULT: The serum calcium level was highest in the follicular phase whereas the serum magnesium level was lowest in the follicular phase; the serum inorganic phosphorus levels was highest in the menstrual phase. CONCLUSION: These variations could be due to the impact of the changing estrogen and progesterone secretion on the parathyroid glands. PMID- 19562051 TI - Interstitial ectopic pregnancy: A rare and difficult clinicosonographic diagnosis. AB - Ectopic pregnancy in the interstitial part of the fallopian tube is a rare event. This condition presents a challenge for clinical as well as radiological diagnosis. Although routine two-dimensional ultrasound can be suggestive, three dimensional ultrasound is highly accurate in diagnosis. Hence, the authors report a rare case of interstitial ectopic pregnancy diagnosed preoperatively by three dimensional ultrasound and managed laparoscopically. PMID- 19562052 TI - Magnetic resonance imaging diagnosis of Mayer-Rokitansky-Kuster-Hauser syndrome. AB - Magnetic resonance imaging (MRI) is the mainstay in the imaging evaluation of Mullerian agenesis, but is not routinely being utilized, particularly in India. Though sagittal MRI clearly demonstrates the absence or hypoplasia of the uterus and the axial images demonstrate the normal ovaries, it is the ability to identify and objectively evaluate other associated anomalies that makes MRI a unique diagnostic modality. It is also noninvasive and has multiplanar capabilities at the same time having a very high soft tissue resolution. We presume it can be used as a comprehensive imaging package for evaluating these patients at one sitting. We report a case of Mullarian agenesis presenting as primary amenorrhea stressing the role and benefits of MRI. PMID- 19562053 TI - Empty follicle syndrome-Still an enigma. AB - Empty follicle syndrome (EFS), although rare with an incidence of 0.2-7%, is a frustrating condition where no oocytes are retrieved in in vitro fertilization (IVF), even though ultrasound and estradiol measurements show the presence of many potential follicles. It is a complex phenomenon that cannot be explained by low bioavailability of human chorionic gonadotrophin alone; neither can it be reliably diagnosed by the measurement of serum beta-human chorionic gonadotrophin (bhCG) on the day of oocyte retrieval (OR), except possibly when the bhCG concentration is very low. Here we report a case who underwent intracytoplasmic sperm injection (ICSI) for her partner's severe oligoasthenozoospermia. Controlled ovarian hyperstimulation (COH) was done in her first cycle of ICSI, using a gonadotrophin-releasing hormone (GnRH) agonist long protocol with follicle-stimulating hormone (FSH) and human menopausal gonadotrophin (HMG). However, as we were unable to retrieve any oocytes, her COH protocol was changed in the subsequent cycle with a successful outcome. PMID- 19562054 TI - Successful pregnancy in a patient with pseudomyxoma peritonei following in-vitro fertilization using donor eggs. AB - Pseudomyxoma peritonei is a rare, chronic relapsing disease with a guarded prognosis. Here, we describe such a case of a young patient presenting with primary infertility, who conceived following in-vitro fertilization with donor egg and had a successful pregnancy outcome. Literature regarding fertility and pregnancy outcome in this condition is reviewed. PMID- 19562055 TI - From the Editor's Desk. PMID- 19562056 TI - Screening of 'Y' chromosome microdeletions in Iranian infertile males. AB - BACKGROUND: It has been hypothesized that microdeletions of Yq may account for a significant proportion of men with infertility. Three nonoverlapping regions, referred to as "azoospermia factors" (AZFa, b, c from proximal to distal Yq) have been defined as spermatogenesis loci and deletions in these regions have been shown to be pathogenically involved in male infertility associated with azoospermia or severe oligospermia. AIMS: Evaluation the frequency of Y chromosome microdeletions in Iranian population. MATERIALS AND METHODS: Fifty infertile men were selected. Semen analysis was done and on the basis of the mean sperm count, all patients were categorized into azoospermia and oligozoospermia, groups. Blood samples were obtained for DNA extraction and chromosomal analysis. Genomic DNA was extracted from blood lymphocytes and amplified by sequence tagged sites-polymerase chain reaction (STS-PCR) method to determine the presence of microdeletions in AZF locus. A total of 34 STS primers including two controls were selected to identify microdeletions of Y chromosome on each subject. RESULTS AND CONCLUSION: 26/50 cases (52%) showed deletion of at least one of the STS Marker. Totally 41 microdeletions was observed. A total of 17 cases (34%) had deletion in one STS. Four oligospermia cases (8%) had deletion in 2 STS site. Three azoospermia cases (6%) had again deletion in 2 STS site, but in different STSs. One case had three deletions in three STS site and finally one individual had seven deletions in AZF locus. The overall frequency of Y chromosome microdeletions observed in the present study was found to be 26/50 (52%). Comparison of our data with the result of other investigators world wide shows that the incidence of Yq microdeletions in Iranian population is much higher than international frequency. Our data agree with other studies regarding microdeletions of AZFc, but for microdeletions of AZFa (14.6%) our results is much higher and differ significantly with many studies. PMID- 19562057 TI - Selective single blastocyst transfer study: 604 cases in 6 years. AB - AIM: To evaluate the credibility of single blastocyst transfer (SBT) method in selected group of patients. SETTINGS AND DESIGN: Retrospective analysis of SBT cases based on computerized data in a private Fertility research centre. MATERIALS AND METHODS: A total of 604 cases of SBTs, done during June 2000 to June 2006, have been analyzed retrospectively to assess the credibility of the method as a method of choice in selective high fertile group of patients. Women between 28 and 42 years have been included in the retrospective analysis, who had adequate number of eggs for fertilization, between 6 and 12. RESULTS AND CONCLUSIONS: Grade I blastocyst transfer resulted in 46.6% of clinical pregnancy and grade II blastocyst transfer resulted in 17.4% of clinical pregnancy rates. Overall pregnancy rate was 64%. Pregnancy loss, as early and late fetal wastages, was 11.06%. PMID- 19562058 TI - Declining semen quality among south Indian infertile men: A retrospective study. AB - BACKGROUND: Male reproductive function has recently attracted increasing attention due to reports on time-related decline in semen quality. Furthermore, regional differences in the semen quality have also been reported. AIM: To investigate the semen quality among large cohort of infertile individuals at a regional level, in terms of the sperm concentration, total sperm motility, sperm morphology and incidence of azoospermia over a period of 13 years. SETTING: University infertility clinic at Kasturba Hospital, Manipal which is a tertiary healthcare centre serving the general population. DESIGN: Retrospective analysis. MATERIALS AND METHODS: This includes a total of 7770 subjects who presented for semen analysis from 1993 to 2005. The data regarding ejaculate volume, sperm density, motility, morphology and the incidence of azoospermia were collected. STATISTICAL ANALYSIS USED: One way analysis of variance (ANOVA), regression analysis and Chi square analysis. RESULTS: The average sperm density among infertile men during 2004-2005 was 26.61 +/- 0.71 millions/mL which was significantly lower than the average sperm density observed in 1993-1994 (38.18 +/- 1.46 millions/mL). Similar trend was also observed for sperm motility (47.14% motile sperms vs. 61.16%) and normal sperm morphology (19.75% vs. 40.51%). Interestingly, the incidence of severe oligospermia (mean sperm density <10 millions/mL) observed in 2002-2005 and 1993-1997 demonstrated a significant inverse relationship (P < 0.001). CONCLUSION: Our study provides the first evidence that the quality of human semen evaluated for infertility is deteriorating in the southern part of the India over the years, probably due to environmental, nutritional, life style or socioeconomic causes. PMID- 19562060 TI - Outcome of twin pregnancies conceived after assisted reproductive techniques. AB - CONTEXT: There is a continuous controversy regarding the obstetric perinatal outcome of twin pregnancies conceived after assisted reproductive techniques (ART). There is an ongoing discussion whether theses parameters may show poorer results as compared to spontaneous conception. AIMS: To evaluate the outcome of multifetal pregnancies and to compare maternal and neonatal complications between spontaneously conceived and assisted reproductive therapy. SETTINGS AND DESIGN: Prospective case-control study. MATERIALS AND METHODS: In this prospective case control study of 2-year duration, obstetric and perinatal outcomes were compared in 36 ART twin pregnancies (Group A) with 138 twins who conceived naturally (Group B). The outcomes were analyzed and used for a comparison between spontaneous and assisted multifetal pregnancies. STATISTICAL ANALYSIS: The continuous variables were analyzed by Student's t-test and categorical variables were analyzed with Fisher's exact test. RESULTS: Pregnancy-related complications like pregnancy-induced hypertension, antepartum hemorrhage, were similar in both groups. Incidence of cesarean section, preterm delivery, and hospital stay was significantly more in Group A vs. Group B, P < 0.001. The newborns in the assisted group had more complications than the spontaneous group; most notable were respiratory distress syndrome, newborn intensive care admission, sepsis, and longer hospital stay (4.8 days vs. 1.6 days, P < 0.001). CONCLUSIONS: Increased rates of cesarean section and preterm delivery are the main reasons for increased obstetric risk in pregnancies conceived through ART. Preterm birth and neonatal prematurity-related complications were the main cause for longer stay in hospital in ART-conceived twins. PMID- 19562059 TI - HLA allele associations in idiopathic recurrent spontaneous abortion patients from India. AB - BACKGROUND: Rejection of semiallogenic foetus in recurrent spontaneous abortion (RSA) has been postulated to be a consequence of genetic and immunological phenomena. AIM: To evaluate the role of human leukocyte antigen (HLA) alleles in RSA in Indian couples. SETTINGS AND DESIGN: A case-control study. MATERIALS AND METHODS: Eighty-one randomly selected couples with unexplained three or more RSAs and a control group of 97 couples with live birth belonging to the same ethnic background, referred to the Gynaecology Department, KEM Hospital were included in the case-control study. Serological HLA A and B typing was done followed by molecular subtypes, defined using PCR-SSOP technique for HLA A, B, and C in 40 couples and DRB1* and DQB1* in 28 couples which were then compared with appropriate case 46 and 88 controls. RESULTS: Serologically A3 (15.43% vs. 4.43%; odds ratio (OR) = 4.34; P = 0.0002) and B17 (25.3% vs. 11.34%; OR = 3.49; P = 0.0001) were increased. Haplotype A1-B17 was significantly increased. Molecular subtyping revealed that A*030102 (11.25% vs. 4.34%; OR = 3.00; P = 0.07), B*5701 (11.25% vs. 1.08%; OR = 13.10; P = 0.003), Cw*120201 (25% vs. 4.34%; OR = 10.50; P = 2.05E-05), HLA DRB1*030101 (17.85% vs. 3.40%; OR = 7.6; P = 0.0001), DRB1*150101 (32.14% vs. 13.63%; OR = 4.8; P = 0.0003), and DQB1*060101 (35.71% vs. 29.34%; OR = 2.3; P = 0.004) were significantly increased in patients. A differential association was noticed when compared with reported world RSA patients. CONCLUSION: The HLA alleles A*030101, B*5701, Cw*120201, DRB1*030101, and DRB1*150101 as well as their associated ancestral haplotype may play a significant role in development of RSA in India. PMID- 19562061 TI - Gonadotrophin releasing hormone antagonist in IVF/ICSI. AB - OBJECTIVE: To study the efficacy of gonadotrophin releasing hormone (GnRH) antagonist in In-vitro-fertilization/Intracytoplasmic sperm injection (IVF/ICSI) cycles. TYPE OF STUDY: Observational study. SETTING: Reproductive Medicine Unit, Christian Medical College Hospital, Vellore, Tamil Nadu. MATERIALS AND METHODS: GnRH antagonists were introduced into our practice in November 2005. Fifty-two women undergoing the antagonist protocol were studied and information gathered regarding patient profile, treatment parameters (total gonadotrophin dosage, duration of treatment, and oocyte yield), and outcomes in terms of embryological parameters (cleavage rates, implantation rates) and clinical pregnancy. These parameters were compared with 121 women undergoing the standard long protocol. The costs between the two groups were also compared. MAIN OUTCOME: Clinical pregnancy rate. RESULTS: The clinical pregnancy rate per embryo transfer in the antagonist group was 31.7% which was comparable to the clinical pregnancy rate in women undergoing the standard long protocol (30.63%). The costs between the two groups were comparable. CONCLUSIONS: GnRH antagonist protocol was found to be effective and comparable to the standard long protocol regimen. In addition it was simple, convenient, and patient friendly. PMID- 19562062 TI - Angular ectopic pregnancy presenting as rupture of lateral wall of the uterus. AB - This is a case report of a 32-year-old woman, being treated for secondary infertility, with history of previous ectopic pregnancy, who presented to the emergency obstetrical room in a state of hypovolemic shock. A diagnosis of ruptured ectopic pregnancy was confirmed in view of history of 14 weeks amenorrhea with a positive urine pregnancy test and positive colpopunture. She was immediately shifted for an emergency exploratory laparotomy. Intraoperatively, the authors were surprised to encounter a right lateral wall rupture uterus and ~14 weeks foetus with the placenta lying freely in the peritoneal cavity. That was suggestive of a right interstitial ectopic which had grown up to 14 weeks, invaded the uterine cavity thus forming an angular ectopic, which ended up as the catastrophic event. The authors here wish to highlight that angular pregnancy is rare but it has catastrophic consequences including maternal mortality. Had the patient presented early, in view of history of previous ectopic, an ultrasonography and color Doppler would have been useful in early detection. And a fertility conserving management in the form of Methotrexate therapy or Selective Uterine artery embolization could have been done. PMID- 19562063 TI - Duplication of the fallopian tube. AB - Hysterosalpingography accurately delineates the uterine and tubal lumen, and hence is routinely performed for the evaluation of infertility.We observed a case of infertility where uterine cavity was normal but fallopian tubes were bifurcated at the ampullary region. Mullerian duct anomalies are reported in literature, but maldevelopment of fallopian tube in isolation is rare. This abnormality can present as infertility, ectopic pregnancy, in association with urinary tract anomalies or as failure of sterilisation method. PMID- 19562064 TI - Heterotopic pregnancy in natural conception. AB - Heterotopic gestation, although common with assisted reproductive techniques, is very rare in natural conception. A high index of suspicion can help in timely diagnosis and appropriate intervention. We report a case of heterotopic pregnancy in a 22-year-old woman presented with hemoperitoneum from ruptured tubal pregnancy with live intrauterine gestation at 10 weeks of amenorrhea, diagnosed on ultrasound examination. PMID- 19562066 TI - From the Editor's Desk. PMID- 19562065 TI - Unruptured ovarian pregnancy following in-vitro fertilization: Missed diagnosis followed by successful laparoscopic management. AB - Ovarian pregnancy after in-vitro fertilization is rare and can be easily missed unless there is a high index of suspicion. Here we present such a case which was missed initially but was later successfully managed laparoscopically. PMID- 19562067 TI - Assessing tubal damage. AB - The fallopian tube plays an important role in the mechanical transport and physiological sustenance of the gametes and early conceptus. Complex and coordinated neuromuscular activity, cilial action and endocrine secretions are required for successful tubal function. Compromised tubal damage can occur after external or internal injury, inhibiting the normal transport of gametes. The overall prognosis for fertility depends principally on the insult and the severity of the tissue damage; hence, assessment of tubal damage plays a major role in predicting occurrence of pregnancy and the likelihood of developing ectopic pregnancy. PMID- 19562068 TI - Comparison of clinical features and health manifestations in lean vs. obese Indian women with polycystic ovarian syndrome. AB - AIMS: To study the prevalence of clinical manifestations in obese and lean polycystic ovarian syndrome (PCOS) women and their health hazards. SETTINGS AND DESIGN: This prospective study was carried out in a tertiary care infertility clinic from 1.7.2005 till 31.12.2007. MATERIALS AND METHODS: These women were diagnosed to have PCOS by the European Society of Human Reproduction and Embryology and the American Society of Reproductive Medicine, Rotterdam 2003 criteria. They were further divided into two groups according to their body mass index (BMI): Group A (n = 300), overweight and obese with BMI >23 and Group B (n = 150), normal weight and lean with BMI 0.85) was similar in both groups, irrespective of body weight (47.7% vs. 38%, P = 0.056, 95% CI=0.06 to +0.18). Comparative data of various health manifestations in lean vs. obese women with POCS [Table 4]. Of the health risk manifestations, hypertension occurred in both groups with a similar frequency (41% vs. 35.5%, P = 0.261, 95% CI=0.03 to +0.15). Group A showed an increased prevalence of IGT (25% vs. 10%, P = 0.000, 95% CI= 0.13-0.29) and type two diabetes mellitus (11.7% vs. 6%, P = 0.000, 95% CI= 0.13-0.29) as compared with group B. endometrial hyperplasia (EH) also showed an increase prevalence in Group A compared with Group B (5.6% vs. 2%, P = 0.055, 95% CI= 0.01-0.08), although not statistically significant. CONCLUSION: PCOS emerges as a clinically heterogeneous condition with increased prevalence of health risks such as hypertension, diabetes and EH. Of these, diabetes and EH appear to be more prevalent in the obese, putting them at a greater risk of morbid problems at a much younger age than the lean ones. PMID- 19562069 TI - The efficacy of metformin and clomiphene citrate combination compared with clomiphene citrate alone for ovulation induction in infertile patients with PCOS. AB - CONTEXT: Low ovulatory and pregnancy rates with clomiphene citrate (CC) in anovulatory polycystic ovarian syndrome (PCOS). AIM: To find out the ovulatory and pregnancy rates in infertile PCOS subjects who receive CC alone and a combination of metformin and CC. SETTING AND DESIGN: A prospective controlled clinical trial conducted in the outpatient department from August 2003 to August 2005. MATERIALS AND METHODS: Twenty-four infertile PCOS women received CC alone at incremental doses of 50 mg up to 150 mg for three cycles and then at a dose of 150 mg for another three cycles (control group). The study group (16 PCOS) received the same dose of CC along with 1500mg of metformin. Ovulation was monitored by transvaginal sonography up to six cycles or till pregnancy occurred. STATISTICAL ANALYSIS: This was carried out using software SSPS, version 10. Fisher's exact test was used to calculate the ovulatory rates. Nine subjects of the control group who failed to conceive with CC had opted for CC and metformin and their ovulatory rate was calculated using statistical software, namely SPSS 15.0, Stata 8.0, MedCalc 9.0.1 and Systat 11.0 using Fischer's exact test. RESULTS: The metformin and clomiphene combination resulted in a significantly higher rate of ovulation (P = 0.0016). The pregnancy rate was 8% with CC and 24% with metformin and CC. The CC failure group also ovulated at a similar rate as that of the study group. CONCLUSIONS: The ovulatory rate and the pregnancy rate with the metformin-CC combination was found to be higher when compared with CC alone. Metformin increased the ovulatory rate in CC failures, also implying increased sensitivity to CC. PMID- 19562070 TI - Blastocyst stage transfer vs cleavage stage embryo transfer. AB - OBJECTIVE: To evaluate the efficacy of blastocyst transfer in comparison with cleavage stage embryo in a similar cohort of women. DESIGN: Retrospective analysis. SETTING: University teaching hospital. MATERIALS AND METHODS: Women aged 35 or less undergoing in vitro fertilization/intracytoplasmic sperm injection between January 2005 and December 2006 were included in the study. When four or more grade 1 embryos were observed on day 3, extended culture till day 5 was undertaken. This policy was compared with a cohort of women who had at least three grade 1 embryos on day 3 and who had undergone a cleavage stage embryo transfer during the time period of January 2002-December 2004. Primary outcome evaluated was implantation rate and clinical pregnancy rate. RESULTS: Group 1 consisted of 50 women who underwent extended culture and blastocyst transfer. Group 2 comprised of 85 women who had cleavage transfer. The implantation rate for embryos transferred in group 1 was significantly higher than that for embryos transferred on day 3 (40.16% vs 11.43%). The clinical pregnancy rate was also significantly better with blastocyst transfer as compared with cleavage stage transfer (62% vs 29.76%). Significantly fewer embryos were required for transfer at the blastocyst stage compared with day 3 transfer (2.54 vs 3.45). CONCLUSION: In selected cases, blastocyst transfer with fewer embryos can be performed with high implantation and clinical pregnancy rates. This policy could lead to a reduction in the incidence of higher-order pregnancies. PMID- 19562071 TI - Plasma protein thiols, ceruloplasmin, C-reactive protein and red blood cell acetylcholinesterase in patients undergoing intrauterine insemination. AB - OBJECTIVE: To estimate acetylcholinesterase (AChE), protein thiols (PT), ceruloplasmin (CP) and C-reactive proteins (CRPs) to assess any change in their levels following intrauterine insemination (IUI). MATERIALS AND METHODS: Forty two patients aged 31 +/- 4.65 years (mean +/- SD) with primary infertility selected for IUI. All of them had induced ovulation with clomiphene citrate 50 mg from day 2 to day 6. After taking the consent, 2 ml of blood was withdrawn before and after 24 h of IUI for biochemical estimations. RESULTS: We observed a significant decrease in plasma CP, PT and RBC AChE (P < 0.001) following IUI compared with the respective pre-procedure levels. Highly sensitive CRP showed a marginal increase after IUI. CONCLUSION: Fluctuations in levels of the above parameters point to their role in the female reproductive system and in the outcome of the IUI. PMID- 19562073 TI - Successful pregnancy following medical management of heterotopic pregnancy. AB - We present a case of sonographic demonstration of quadruplet heterotopic pregnancy consisting of twin intrauterine (IU) pregnancy and a twin adnexal pregnancy after ovulation induction (OI) with clomiphene citrate (CC) and timed intercourse (TI). Both heterotopic pregnancy and spontaneous twinning are frequent after OI, this combination although extremely rare must be kept in mind. The role of early transvaginal sonography and serum beta human chorionic gonadotrophin after missed periods helps in early diagnosis. It gives us an opportunity for medical management, saving the patient the agony of surgery along with loss of pregnancy. The management of heterotopic pregnancy is controversial. This patient did not have a viable IU pregnancy and both the sacs in the adnexa were small. Thus, we treated her successfully by medical management with systemic methotrexate, with regular follow-up. This patient successfully conceived after 6 months with OI and TI, with ovulation occurring from the same side of the previous ectopic. She had a viable IU gestation corresponding to 12 weeks. PMID- 19562072 TI - Tomato (Lycopersicon esculentum) prevents lead-induced testicular toxicity. AB - BACKGROUND: Lead, an example of heavy metals, has, for decades, being known for its adverse effects on various body organs and systems such that their functions are compromised. AIM: In the present study, the ability of lead to adversely affect the male reproductive system was investigated and tomato (Lycopersicon esculentum: Source of antioxidants) paste (TP) was administered orally to prevent the adverse effects of Pb. MATERIALS AND METHODS: Fifteen Sprague Dawley rats, randomised into three groups (n = 5), were used for this study. Animals in Group A served as the control and were drinking distilled water. Animals in Groups B and C were drinking 1% Pb (II) acetate (LA). Group C animals were, in addition to drinking LA, treated with 1.5 ml of TP/day. All treatments were for 8 weeks. STATISTICAL ANALYSIS USED: A Mann-Whitney U-test was used to analyse the results obtained. RESULTS: The obtained results showed that Pb caused a significant reduction in the testicular weight, sperm count, life-death ratio, sperm motility, normal sperm morphology, and plasma and tissue superoxide dismutase and catalase activity, but a significant increase in plasma and tissue malondialdehyde concentration. But, Pb did not cause any significant change in the serum testosterone level. TP, however, significantly reduced these adverse effects of Pb. CONCLUSION: These findings lead to the conclusion that TP significantly lowered the adverse effects of Pb exposure on the kidney as well as Pb-induced oxidative stress. PMID- 19562074 TI - Successful birth of the first frozen oocyte baby in India. AB - We report the first pregnancy and birth in India after the transfer of embryos generated from frozen- thawed oocytes. A 29-year-old woman with previous bad obstetric history and an abnormal karyotype, necessitating donor oocyte programme. Embryos were generated by microinjection of frozen-thawed sperms into thawed human oocytes (intracytoplasmic sperm injection). This resulted in an healthy male baby with a birth weight of 2.54 kg which was born by cesarean section at 35-36 weeks of gestation with normal follow-up. Thus oocyte cryopreservation can be performed with reproducible success leading to a viable offspring. PMID- 19562075 TI - Successful management of post-in-vitro fertilization cervical heterotropic pregnancy. AB - A case of post-in-vitro fertilization (IVF) cervical and intrauterine heterotropic pregnancy with cardiac activity in both embryos is presented. It was diagnosed in the 7(th) week of gestation by ultrasonography and cervical pregnancy was treated conservatively with intrathoracic administration of potassium chloride under transvaginal ultrasound guidance with regression of trophoblastic tissue. The intrauterine pregnancy continued and cesarean section was performed in the 31(st) week due to absent end diastolic umbilical artery fetal blood flow. A live male baby was delivered. PMID- 19562076 TI - Gene order phylogeny and the evolution of methanogens. AB - Methanogens are a phylogenetically diverse group belonging to Euryarchaeota. Previously, phylogenetic approaches using large datasets revealed that methanogens can be grouped into two classes, "Class I" and "Class II". However, some deep relationships were not resolved. For instance, the monophyly of "Class I" methanogens, which consist of Methanopyrales, Methanobacteriales and Methanococcales, is disputable due to weak statistical support. In this study, we use MSOAR to identify common orthologous genes from eight methanogen species and a Thermococcale species (outgroup), and apply GRAPPA and FastME to compute distance-based gene order phylogeny. The gene order phylogeny supports two classes of methanogens, but it differs from the original classification of methanogens by placing Methanopyrales and Methanobacteriales together with Methanosarcinales in Class II rather than with Methanococcales. This study suggests a new classification scheme for methanogens. In addition, it indicates that gene order phylogeny can complement traditional sequence-based methods in addressing taxonomic questions for deep relationships. PMID- 19562078 TI - A principal component analysis of 39 scientific impact measures. AB - BACKGROUND: The impact of scientific publications has traditionally been expressed in terms of citation counts. However, scientific activity has moved online over the past decade. To better capture scientific impact in the digital era, a variety of new impact measures has been proposed on the basis of social network analysis and usage log data. Here we investigate how these new measures relate to each other, and how accurately and completely they express scientific impact. METHODOLOGY: We performed a principal component analysis of the rankings produced by 39 existing and proposed measures of scholarly impact that were calculated on the basis of both citation and usage log data. CONCLUSIONS: Our results indicate that the notion of scientific impact is a multi-dimensional construct that can not be adequately measured by any single indicator, although some measures are more suitable than others. The commonly used citation Impact Factor is not positioned at the core of this construct, but at its periphery, and should thus be used with caution. PMID- 19562077 TI - Dll1 haploinsufficiency in adult mice leads to a complex phenotype affecting metabolic and immunological processes. AB - BACKGROUND: The Notch signaling pathway is an evolutionary conserved signal transduction pathway involved in embryonic patterning and regulation of cell fates during development and self-renewal. Recent studies have demonstrated that this pathway is integral to a complex system of interactions, involving as well other signal transduction pathways, and implicated in distinct human diseases. Delta-like 1 (Dll1) is one of the known ligands of the Notch receptors. The role of the Notch ligands is less well understood. Loss-of-function of Dll1 leads to embryonic lethality, but reduction of Delta-like 1 protein levels has not been studied in adult stage. METHODOLOGY/PRINCIPAL FINDINGS: Here we present the haploinsufficient phenotype of Dll1 and a missense mutant Dll1 allele (Dll1(C413Y)). Haploinsufficiency leads to a complex phenotype with several biological processes altered. These alterations reveal the importance of Dll1 mainly in metabolism, energy balance and in immunology. The animals are smaller, lighter, with altered fat to lean ratio and have increased blood pressure and a slight bradycardia. The animals have reduced cholesterol and triglyceride levels in blood. At the immunological level a subtle phenotype is observed due to the effect and fine-tuning of the signaling network at the different levels of differentiation, proliferation and function of lymphocytes. Moreover, the importance of the proteolytic regulation of the Notch signaling network emphasized. CONCLUSIONS/SIGNIFICANCE: In conclusion, slight alterations in one player of Notch signaling alter the entire organism, emphasizing the fine-tuning character of this pathway in a high number of processes. PMID- 19562079 TI - More than 9,000,000 unique genes in human gut bacterial community: estimating gene numbers inside a human body. AB - BACKGROUND: Estimating the number of genes in human genome has been long an important problem in computational biology. With the new conception of considering human as a super-organism, it is also interesting to estimate the number of genes in this human super-organism. PRINCIPAL FINDINGS: We presented our estimation of gene numbers in the human gut bacterial community, the largest microbial community inside the human super-organism. We got 552,700 unique genes from 202 complete human gut bacteria genomes. Then, a novel gene counting model was built to check the total number of genes by combining culture-independent sequence data and those complete genomes. 16S rRNAs were used to construct a three-level tree and different counting methods were introduced for the three levels: strain-to-species, species-to-genus, and genus-and-up. The model estimates that the total number of genes is about 9,000,000 after those with identity percentage of 97% or up were merged. CONCLUSION: By combining completed genomes currently available and culture-independent sequencing data, we built a model to estimate the number of genes in human gut bacterial community. The total number of genes is estimated to be about 9 million. Although this number is huge, we believe it is underestimated. This is an initial step to tackle this gene counting problem for the human super-organism. It will still be an open problem in the near future. The list of genomes used in this paper can be found in the supplementary table. PMID- 19562080 TI - Cool sex? Hibernation and reproduction overlap in the echidna. AB - During hibernation there is a slowing of all metabolic processes, and thus it is normally considered to be incompatible with reproduction. In Tasmania the egg laying mammal, the echidna (Tachyglossus aculeatus) hibernates for several months before mating in mid-winter, and in previous studies we observed males with females that were still hibernating. We monitored the reproductive activity of radio-tracked echidnas by swabbing the reproductive tract for sperm while external temperature loggers provided information on the timing of hibernation. Additional information was provided by camera traps and ultrasound imaging. More than a third of the females found in mating groups were torpid, and the majority of these had mated. Some females re-entered deep torpor for extended periods after mating. Ultrasound examination showed a developing egg in the uterus of a female that had repeatedly re-entered torpor. The presence of fresh sperm in cloacal swabs taken from this female on three occasions after her presumed date of fertilization indicated she mated several times after being fertilized. The mating of males with torpid females is the result of extreme competition between promiscuous males, while re-entry into hibernation by pregnant females could improve the possibility of mating with a better quality male. PMID- 19562082 TI - Uses and misuses of progress curve analysis in enzyme kinetics. AB - Progress curve analysis is a convenient tool for the characterization of enzyme action: a single reaction mixture provides multiple experimental measured points for continuously varying amounts of substrates and products with exactly the same enzyme and modulator concentrations. The determination of kinetic parameters from the progress curves, however, requires complex mathematical evaluation of the time-course data. Some freely available programs (e.g. FITSIM, DYNAFIT) are widely applied to fit kinetic parameters to user-defined enzymatic mechanisms, but users often overlook the stringent requirements of the analytic procedures for appropriate design of the input experiments. Flaws in the experimental setup result in unreliable parameters with consequent misinterpretation of the biological phenomenon under study. The present commentary suggests some helpful mathematical tools to improve the analytic procedure in order to diagnose major errors in concept and design of kinetic experiments. PMID- 19562081 TI - Intragraft selection of the T cell receptor repertoire by class I MHC sequences in tolerant recipients. AB - BACKGROUND: Allograft tolerance of ACI (RT1(a)) recipients to WF (RT1(u)) hearts can be induced by allochimeric class I MHC molecules containing donor-type (RT1A(u)) immunogenic epitopes displayed on recipient-type (RT1A(a)) sequences. Here, we sought the mechanisms by which allochimeric sequences may affect responding T cells through T cell receptor (TCA) repertoire restriction. METHODOLOGY/PRINCIPAL FINDINGS: The soluble [alpha(1h) (u)]-RT1.A(a) allochimeric molecule was delivered into ACI recipients of WF hearts in the presence of sub therapeutic dose of cyclosporine (CsA). The TCR Vbeta spectrotyping of the splenocytes and cardiac allografts showed that the Vbeta gene families were differentially expressed within the TCR repertoire in allochimeric- or high-dose CsA-treated tolerant recipients at day +5 and +7 of post-transplantation. However, at day 30 of post-transplantation the allochimeric molecule-treated rats showed the restriction of TCR repertoire with altered dominant size peaks representing preferential clonal expansion of Vbeta7, Vbeta11, Vbeta13, Vbeta 14, and Vbeta15 genes. Moreover, we found a positive correlation between the alteration of Vbeta profile, restriction of TCR repertoire, and the establishment of allograft tolerance. CONCLUSIONS: Our findings indicate that presentation of allochimeric MHC class I sequences that partially mimic donor and recipient epitopes may induce unique tolerant state by selecting alloresponsive Vbeta genes. PMID- 19562084 TI - Characterizing the dynamic frequency structure of fast oscillations in the rodent hippocampus. AB - Fast oscillations or "ripples" are found in the local field potential (LFP) of the rodent hippocampus during awake and sleep states. Ripples have been found to correlate with memory related neural processing, however, the functional role of the ripple has yet to be fully established. We applied a Kalman smoother based estimator of instantaneous frequency (iFreq) and frequency modulation (FM) to ripple oscillations recorded in-vivo from region CA1 of the rat and mouse hippocampus during slow wave sleep. We found that (1) ripples exhibit stereotypical frequency dynamics that are consistent in the rat and mouse, (2) instantaneous frequency information may be used as an additional dimension in the classification of ripple events, and (3) the instantaneous frequency structure of ripples may be used to improve the detection of ripple events by reducing Type I and Type II errors. Based on our results, we propose that high temporal and spectral resolution estimates of frequency dynamics may be used to help elucidate the mechanisms of ripple generation and memory related processing. PMID- 19562085 TI - Establishing a novel modeling tool: a python-based interface for a neuromorphic hardware system. AB - Neuromorphic hardware systems provide new possibilities for the neuroscience modeling community. Due to the intrinsic parallelism of the micro-electronic emulation of neural computation, such models are highly scalable without a loss of speed. However, the communities of software simulator users and neuromorphic engineering in neuroscience are rather disjoint. We present a software concept that provides the possibility to establish such hardware devices as valuable modeling tools. It is based on the integration of the hardware interface into a simulator-independent language which allows for unified experiment descriptions that can be run on various simulation platforms without modification, implying experiment portability and a huge simplification of the quantitative comparison of hardware and simulator results. We introduce an accelerated neuromorphic hardware device and describe the implementation of the proposed concept for this system. An example setup and results acquired by utilizing both the hardware system and a software simulator are demonstrated. PMID- 19562083 TI - EGFR Inhibition in the Treatment of Non-Small Cell Lung Cancer. AB - Epidermal growth factor receptor (EGFR) inhibitors have introduced the concept of targeted therapy to the treatment of non-small cell lung cancer (NSCLC). These agents appear most effective in patients with tumors that are highly dependent on EGFR signaling pathways, a population that disproportionately includes females, nonsmokers, individuals of East Asian origin, and patients with adenocarcinoma histology. Currently available EGFR-inhibiting drugs include the tyrosine kinase inhibitors (TKIs) erlotinib, gefitinib, and lapatinib, which are administered orally and interfere with the intracellular tyrosine kinase domain, and the monoclonal antibodies (mAbs) cetuximab and panitumumab, which are administered intravenously and interfere with extracellular ligand binding. While the use of EGFR TKIs as monotherapy prolongs survival in metastatic NSCLC, they have demonstrated no benefit when added to conventional, cytotoxic chemotherapy. In contrast, the anti-EGFR mAb cetuximab appears most effective when combined with chemotherapy or radiation. Despite dramatic initial responses to treatment in some cases, NSCLC eventually becomes resistant to EGFR inhibition. Possible mechanisms include secondary mutations that interfere with drug binding, oncogenic pathways driven by other receptor tyrosine kinases, and independent activity of downstream signaling molecules. Efforts to overcome such resistance include irreversibly binding EGFR TKIs, multi-targeted TKIs, and combinations with chemotherapy, radiation, and other targeted therapies. PMID- 19562086 TI - Daily rhythm of melanopsin-expressing cells in the mouse retina. AB - In addition to some other functions, melanopsin-expressing retinal ganglion cells (RGCs) constitute the principal mediators of the circadian photoentrainment, a process by which the suprachiasmatic nucleus (the central clock of mammals), adjusts daily to the external day/night cycle. In the present study these RGCs were immunohistochemically labelled using a specific polyclonal antiserum raised against mouse melanopsin. A daily oscillation in the number of immunostained cells was detected in mice kept under a light / dark (LD) cycle. One hour before the lights were on (i.e., the end of the night period) the highest number of immunopositive cells was detected while the lowest was seen 4 h later (i.e., within the first hours of the light period). This finding suggests that some of the melanopsin-expressing RGCs "turn on" and "off" during the day/night cycle. We have also detected that these daily variations already occur in the early postnatal development, when the rod/cone photoreceptor system is not yet functional. Two main melanopsin-expressing cell subpopulations could be found within the retina: M1 cells showed robust dendritic arborization within the OFF sublamina of the inner plexiform layer (IPL), whilst M2 cells had fine dendritic processes within the ON sublamina of the IPL. These two cell subpopulations also showed different daily oscillations throughout the LD cycle. In order to find out whether or not the melanopsin rhythm was endogenous, other mice were maintained in constant darkness for 6 days. Under these conditions, no defined rhythm was detected, which suggests that the daily oscillation detected either is light dependent or is gradually lost under constant conditions. This is the first study to analyze immunohistochemically the daily oscillation of the number of melanopsin-expressing cells in the mouse retina. PMID- 19562087 TI - Cannabinoid receptor activation reverses kainate-induced synchronized population burst firing in rat hippocampus. AB - Cannabinoids have been shown to possess anticonvulsant properties in whole animal models of epilepsy. The present investigation sought to examine the effects of cannabinoid receptor activation on kainic acid (KA)-induced epileptiform neuronal excitability. Under urethane anesthesia, acute KA treatment (10 mg kg(-1), i.p.) entrained the spiking mode of simultaneously recorded neurons from random firing to synchronous bursting (% change in burst rate). Injection of the high-affinity cannabinoid agonist (-)-11-hydroxy-8-tetrahydrocannabinol-dimethyl-heptyl (HU210, 100 mug kg(-1), i.p.) following KA markedly reduced the burst frequency (% decrease in burst frequency) and reversed synchronized firing patterns back to baseline levels. Pre-treatment with the central cannabinoid receptor (CB1) antagonist N-piperidino-5-(4-clorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-3 pyrazole-carboxamide (rimonabant, SR141716A 3 mg kg(-1), i.p.) completely prevented the actions of HU210. The present results indicate that cannabinoids exert their antiepileptic effects by impeding pathological synchronization of neuronal networks in the hippocampus. PMID- 19562088 TI - Inter- and intra-individual covariations of hemodynamic and oscillatory gamma responses in the human cortex. AB - The time course of local field potentials (LFPs) displaying typical discharge frequencies in the gamma frequency range highly correlates with the blood oxygen level dependent (BOLD) signal in response to rotating checkerboard stimuli in animals. In humans, oscillatory gamma-band responses (GBRs) show strong inter individual variations in frequency and amplitude but considerable intra individual reliability indicating that individual gamma activity reflects a personal trait. While the functional role of these GBRs is still debated, investigations combining electroencephalography-functional magnetic resonance imaging (EEG-fMRI) measurements provide a tool to obtain further insights into the underlying functional architecture of the human brain and will shed light onto the understanding of the dynamic relation between the BOLD signal and the properties of the electrical activity recorded on the scalp. We investigated the relation between the hemodynamic response and evoked gamma-band response (eGBR) to visual stimulation. We tested the hypothesis that the amplitude of human eGBRs and BOLD responses covary intra-individually as a function of stimulation as well as inter-individually as a function of gamma-trait. Seventeen participants performed visual discrimination tasks during separate EEG and fMRI recordings. Results revealed that visual stimuli that evoked high GBRs also elicited strong BOLD responses in the human V1/V2 complex. Furthermore, inter-individual variations of BOLD responses to visual stimuli in the bilateral primary (Area 17) and secondary (Area V5/MT) visual cortex and the right hippocampal formation were correlated with the individual gamma-trait of the subjects. The present study further supports the notion that neural oscillations in the gamma frequency range are involved in the cascade of neural processes that underlie the hemodynamic responses measured with fMRI. PMID- 19562089 TI - Comparison of the efficacy of two anticonvulsants, phenytoin and valproate to improve PCP and d-amphetamine induced deficits in a reversal learning task in the rat. AB - Recent studies in our laboratory have shown that PCP (phencyclidine) and d amphetamine induce a cognitive deficit in rats, in a paradigm of potential relevance for the pathology of schizophrenia. Atypical, but not classical antipsychotics and the anticonvulsant, lamotrigine have been shown to prevent a selective reversal learning deficit induced by PCP. In contrast, only haloperidol reversed the d-amphetamine-induced deficit. The present study aimed to explore the ability of two anticonvulsants with differing mechanism of action, valproate and phenytoin to attenuate the cognitive deficits induced by PCP and d amphetamine in the reversal learning paradigm. PCP at 1.5 mg/kg and d-amphetamine at 0.5 mg/kg both produced a selective and significant reduction in performance of the reversal phase with no effect on the initial phase of the task in female hooded Lister rats. Valproate (25-200 mg/kg) and phenytoin (25-50 mg/kg) had no effect on performance when administered alone. Valproate (100-200 mg/kg), whose principle action is thought to be the enhancement of GABA transmission, was unable to prevent the cognitive deficit induced by either PCP or d-amphetamine. Conversely, phenytoin (50 mg/kg), a use-dependent sodium channel inhibitor, significantly prevented the deficit induced by PCP, but not d-amphetamine. These results add to our earlier work with lamotrigine, and suggest that sodium channel blockade may be a mechanism by which some anticonvulsant drugs can prevent the PCP-induced deficit. These data have implications for the use of anticonvulsant drugs in the treatment of cognitive or psychotic disorders. PMID- 19562090 TI - Design and adsorption of modular engineered proteins to prepare customized, neuron-compatible coatings. AB - Neural prosthetic implants are currently being developed for the treatment and study of both peripheral and central nervous system disorders. Effective integration of these devices upon implantation is a critical hurdle to achieving function. As a result, much attention has been directed towards the development of biocompatible coatings that prolong their in vivo lifespan. In this work, we present a novel approach to fabricate such coatings, which specifically involves the use of surface-adsorbed, nanoscale-designed protein polymers to prepare reproducible, customized surfaces. A nanoscale modular design strategy was employed to synthesize six engineered, recombinant proteins intended to mimic aspects of the extracellular matrix proteins fibronectin, laminin, and elastin as well as the cell-cell adhesive protein neural cell adhesion molecule. Physical adsorption isotherms were experimentally determined for these engineered proteins, allowing for direct calculation of the available ligand density present on coated surfaces. As confirmation that ligand density in these engineered systems impacts neuronal cell behavior, we demonstrate that increasing the density of fibronectin-derived RGD ligands on coated surfaces while maintaining uniform protein surface coverage results in enhanced neurite extension of PC-12 cells. Therefore, this engineered protein adsorption approach allows for the facile preparation of tunable, quantifiable, and reproducible surfaces for in vitro studies of cell-ligand interactions and for potential application as coatings on neural implants. PMID- 19562091 TI - Temporal discounting and inter-temporal choice in rhesus monkeys. AB - Humans and animals are more likely to take an action leading to an immediate reward than actions with delayed rewards of similar magnitudes. Although such devaluation of delayed rewards has been almost universally described by hyperbolic discount functions, the rate of this temporal discounting varies substantially among different animal species. This might be in part due to the differences in how the information about reward is presented to decision makers. In previous animal studies, reward delays or magnitudes were gradually adjusted across trials, so the animals learned the properties of future rewards from the rewards they waited for and consumed previously. In contrast, verbal cues have been used commonly in human studies. In the present study, rhesus monkeys were trained in a novel inter-temporal choice task in which the magnitude and delay of reward were indicated symbolically using visual cues and varied randomly across trials. We found that monkeys could extract the information about reward delays from visual symbols regardless of the number of symbols used to indicate the delay. The rate of temporal discounting observed in the present study was comparable to the previous estimates in other mammals, and the animal's choice behavior was largely consistent with hyperbolic discounting. Our results also suggest that the rate of temporal discounting might be influenced by contextual factors, such as the novelty of the task. The flexibility furnished by this new inter-temporal choice task might be useful for future neurobiological investigations on inter-temporal choice in non-human primates. PMID- 19562093 TI - Intramolecular thermal allenyne [2 + 2] cycloadditions; facile construction of the 5-6-4 ring core of sterpurene. AB - A variety of 1-allenyl-2-propargyl-substituted cyclopentanol derivatives were found to undergo facile intramolecular microwave-assisted 2+2 alleneyne cycloaddition reactions to generate tricyclic 5-6-4 ring systems present in the sterpurenes. PMID- 19562094 TI - The azido acid approach to beta-peptides: parallel synthesis of a tri-beta peptide library by fluorous tagging. AB - A small tri-beta-peptide library was prepared starting from three enantio- and diastereopure azido acids. Fluorous tagging followed by two cycles of azide reduction, fluorous solid phase extraction (f-SPE), peptide coupling with the original azido acids and f-SPE provided 27 protected azido peptides. Reduction and HPLC purification provided 25 of the 27 targeted tri-beta-peptides in acceptable yields and excellent purities. PMID- 19562092 TI - Low and High Gamma Oscillations in Rat Ventral Striatum have Distinct Relationships to Behavior, Reward, and Spiking Activity on a Learned Spatial Decision Task. AB - Local field potential (LFP) oscillations in the brain reflect organization thought to be important for perception, attention, movement, and memory. In the basal ganglia, including dorsal striatum, dysfunctional LFP states are associated with Parkinson's disease, while in healthy subjects, dorsal striatal LFPs have been linked to decision-making processes. However, LFPs in ventral striatum have been less studied. We report that in rats running a spatial decision task, prominent gamma-50 (45-55 Hz) and gamma-80 (70-85 Hz) oscillations in ventral striatum had distinct relationships to behavior, task events, and spiking activity. Gamma-50 power increased sharply following reward delivery and before movement initiation, while in contrast, gamma-80 power ramped up gradually to reward locations. Gamma-50 power was low and contained little structure during early learning, but rapidly developed a stable pattern, while gamma-80 power was initially high before returning to a stable level within a similar timeframe. Putative fast-spiking interneurons (FSIs) showed phase, firing rate, and coherence relationships with gamma-50 and gamma-80, indicating that the observed LFP patterns are locally relevant. Furthermore, in a number of FSIs such relationships were specific to gamma-50 or gamma-80, suggesting that partially distinct FSI populations mediate the effects of gamma-50 and gamma-80. PMID- 19562095 TI - Design maps for failure of all-ceramic layer structures in concentrated cyclic loading. AB - A study is made of the competition between failure modes in ceramic-based bilayer structures joined to polymer-based substrates, in simulation of dental crown-like structures with a functional but weak "veneer" layer bonded onto a strong "core" layer. Cyclic contact fatigue tests are conducted in water on model flat systems consisting of glass plates joined to glass, sapphire, alumina or zirconia support layers glued onto polycarbonate bases. Critical numbers of cycles to take each crack mode to failure are plotted as a function of peak contact load on failure maps showing regions in which each fracture mode dominates. In low-cycle conditions, radial and outer cone cracks are competitive in specimens with alumina cores, and outer cone cracks prevail in specimens with zirconia cores; in high-cycle conditions, inner cone cracks prevail in all cases. The roles of other factors, e.g. substrate modulus, layer thickness, indenter radius and residual stresses from specimen preparation, are briefly considered. PMID- 19562096 TI - Characterization of a High Efficiency, Ultrashort Pulse Shaper Incorporating a Reflective 4096-Element Spatial Light Modulator. AB - We demonstrate pulse shaping via arbitrary phase modulation with a reflective, 1*4096 element, liquid crystal spatial light modulator (SLM). The unique construction of this device provides a very high efficiency when the device is used for phase modulation only in a prism based pulse shaper, namely 85%. We also present a single shot characterization of the SLM in the spatial domain and a single shot characterization of the pulse shaper in the spectral domain. These characterization methods provide a detailed picture of how the SLM modifies the spectral phase of an ultrashort pulse. PMID- 19562097 TI - Decision Making in Fuzzy Discrete Event Systems1. AB - The primary goal of the study presented in this paper is to develop a novel and comprehensive approach to decision making using fuzzy discrete event systems (FDES) and to apply such an approach to real-world problems. At the theoretical front, we develop a new control architecture of FDES as a way of decision making, which includes a FDES decision model, a fuzzy objective generator for generating optimal control objectives, and a control scheme using both disablement and enforcement. We develop an online approach to dealing with the optimal control problem efficiently. As an application, we apply the approach to HIV/AIDS treatment planning, a technical challenge since AIDS is one of the most complex diseases to treat. We build a FDES decision model for HIV/AIDS treatment based on expert's knowledge, treatment guidelines, clinic trials, patient database statistics, and other available information. Our preliminary retrospective evaluation shows that the approach is capable of generating optimal control objectives for real patients in our AIDS clinic database and is able to apply our online approach to deciding an optimal treatment regimen for each patient. In the process, we have developed methods to resolve the following two new theoretical issues that have not been addressed in the literature: (1) the optimal control problem has state dependent performance index and hence it is not monotonic, (2) the state space of a FDES is infinite. PMID- 19562098 TI - Segmentation of Textures Defined on Flat vs. Layered Surfaces using Neural Networks: Comparison of 2D vs. 3D Representations. AB - Texture boundary detection (or segmentation) is an important capability in human vision. Usually, texture segmentation is viewed as a 2D problem, as the definition of the problem itself assumes a 2D substrate. However, an interesting hypothesis emerges when we ask a question regarding the nature of textures: What are textures, and why did the ability to discriminate texture evolve or develop? A possible answer to this question is that textures naturally define physically distinct (i.e., occluded) surfaces. Hence, we can hypothesize that 2D texture segmentation may be an outgrowth of the ability to discriminate surfaces in 3D. In this paper, we conducted computational experiments with artificial neural networks to investigate the relative difficulty of learning to segment textures defined on flat 2D surfaces vs. those in 3D configurations where the boundaries are defined by occluding surfaces and their change over time due to the observer's motion. It turns out that learning is faster and more accurate in 3D, very much in line with our expectation. Furthermore, our results showed that the neural network's learned ability to segment texture in 3D transfers well into 2D texture segmentation, bolstering our initial hypothesis, and providing insights on the possible developmental origin of 2D texture segmentation function in human vision. PMID- 19562099 TI - Surface Plasmon Coupled Phosphorescence (SPCP). AB - We report the first observation of surface plasmon coupled phosphorescence (SPCP) for PtOEP (2,3,7,8,12,17,18-octaethyl-21H,23H-porphyrin platinum II) immobilized in polyvinyl chloride (PVC) films on continuous gold films of 47 nm thickness. We confirm the observation of SPCP by comparing the spectra and lifetimes for plasmon coupled phosphorescence with the free space isotropic phosphorescence emission. Due to the directional emission inherent to plasmon coupled phosphorescence, we believe that SPCP can facilitate the study of phosphorescence in biological applications, such as protein dynamics and alkaline phosphatase studies, whose signal intensities are inherently weak and nearly always isotropic. PMID- 19562100 TI - Synthesis of protected norcysteines for SPPS compatible with Fmoc-strategy. AB - We report the synthesis of racemic Alloc-Ncy(Tmob)-OH, the resolution of its methyl ester, and demonstrate its application to form a norcystine bridge in octreotide-amide using the Fmoc-strategy on solid phase. N-Alloc and S-Tmob protections of norcysteine (Ncy) were found to be a preferred choice for Fmoc strategy over three other protected norcysteines synthesized i.e. Fmoc-Ncy(tBu) OH, Alloc-Ncy(tBu)-OH and Alloc-Ncy(Trt)-OH. PMID- 19562101 TI - Implications of National Suicide Trends for Social Work Practice with Black Youth. AB - Although homicide is the leading cause of death for African-Americans aged 15-24, suicide is silently claiming the lives of many African-American youth, males in particular. Given the disproportionate number of African-American adolescents in many of the primary human service institutions, it is important to increase social workers' understanding of the nature and trends in self-destructive behaviors of this population. This paper presents the descriptive epidemiological trend data on African-American adolescent suicide completion and parasuicidal behavior, reviews current explanatory hypotheses, highlights important risk and protective factors, and outlines several culturally-congruent practice guidelines for working with suicidal African-Americans adolescents. PMID- 19562102 TI - Experimental Evidence for Space-Charge Effects between Ions of the Same Mass-to Charge in Fourier-Transform Ion Cyclotron Resonance Mass Spectrometry. AB - It is often stated that ions of the same mass-to-charge do not induce space charge frequency shifts among themselves in an ion cyclotron resonance mass spectrometry measurement. Here, we demonstrate space-charge induced frequency shifts for ions of a single mass-to-charge. The monoisotopic atomic ion, Cs(+), was used for this study. The measured frequency is observed to decrease linearly with an increase in the number of ions, as has been reported previously for space charge effects between ions of different mass-to-charge. The frequency shift between ions of the same m/z value are compared to that induced between ions of different m/z value, and is found to be 7.5 times smaller. Control experiments were performed to ensure that the observed space-charge effects are not artifacts of the measurement or of experimental design. The results can be rationalized by recognizing that the electric forces between ions in a magnetic field conform to the weak form of the Newton's third law, where the action and reaction forces do not cancel exactly. PMID- 19562107 TI - Darwinian chemistry: towards the synthesis of a simple cell. AB - The total synthesis of a simple cell is in many ways the ultimate challenge in synthetic biology. Outlined eight years ago in a visionary article by Szostak et al. (J. W. Szostak, D. P. Bartel and P. L. Luisi, Nature, 2001, 409, 387), the chances of success seemed remote. However, recent progress in nucleic acid chemistry, directed evolution and membrane biophysics have brought the prospect of a simple synthetic cell with life-like properties such as growth, division, heredity and evolution within reach. Success in this area will not only revolutionize our understanding of abiogenesis but provide a fertile test-bed for models of prebiotic chemistry and early evolution. Last but not least, a robust "living" protocell may provide a versatile and safe chassis for embedding synthetic devices and systems. PMID- 19562108 TI - Decoding biological principles using gene circuits. AB - A major flavor of synthetic biology is the creation of artificial gene circuits to perform user-defined tasks. One aspect of this area is to realize ever increasingly more complicated circuit behavior. Such efforts have led to the identification and evaluation of design strategies that enable robust control of dynamics in single cells and in cell populations. On the other hand, there is increasing emphasis on using artificial systems programmed by simple circuits to explore fundamental biological questions of broad significance. PMID- 19562106 TI - Biocomputers: from test tubes to live cells. AB - Biocomputers are man-made biological networks whose goal is to probe and control biological hosts--cells and organisms--in which they operate. Their key design features, informed by computer science and engineering, are programmability, modularity and versatility. While still a work in progress, biocomputers will eventually enable disease diagnosis and treatment with single-cell precision, lead to "designer" cell functions for biotechnology, and bring about a new generation of biological measurement tools. This review describes the intellectual foundation of the "biocomputer" concept as well as surveys the state of the art in the field. PMID- 19562109 TI - Synthetic biology: exploring and exploiting genetic modularity through the design of novel biological networks. AB - Synthetic biology has been used to describe many biological endeavors over the past thirty years--from designing enzymes and in vitro systems, to manipulating existing metabolisms and gene expression, to creating entirely synthetic replicating life forms. What separates the current incarnation of synthetic biology from the recombinant DNA technology or metabolic engineering of the past is an emphasis on principles from engineering such as modularity, standardization, and rigorously predictive models. As such, synthetic biology represents a new paradigm for learning about and using biological molecules and data, with applications in basic science, biotechnology, and medicine. This review covers the canonical examples as well as some recent advances in synthetic biology in terms of what we know and what we can learn about the networks underlying biology, and how this endeavor may shape our understanding of living systems. PMID- 19562110 TI - Advancing high-throughput gene synthesis technology. AB - The emerging field of synthetic biology is generating insatiable demands for synthetic genes, which far exceed existing gene synthesis capabilities. This review discusses the current methods of chemical DNA synthesis and gene assembly, as well as the latest engineering tools, technologies and trends which could potentially lead to breakthroughs in the development of accurate, low-cost and high-throughput gene synthesis technology. The capability of generating unlimited supplies of DNA molecules of any sequence or size will transform biomedical research in the near future. PMID- 19562111 TI - Engineering and exploiting protein assemblies in synthetic biology. AB - Many biologically relevant structures are formed by the self-assembly of identical protein units. Examples include virus capsids or cytoskeleton components. Synthetic biology can harness these bottom-up assemblies and expand their scope for applications in cell biology and biomedicine. Nanobiotechnology and materials science also stand to gain from assemblies with unique nanoscale periodicity. In these disciplines, the soft scaffolds can serve as templates to produce new metallic or inorganic materials of predefined dimensions. This review article describes how the structure and function of biological assemblies has inspired researchers to develop engineered systems with designed properties for new biomolecular applications. PMID- 19562112 TI - Towards the automated engineering of a synthetic genome. AB - The development of the technology to synthesize new genomes and to introduce them into hosts with inactivated wild-type chromosome opens the door to new horizons in synthetic biology. Here it is of outmost importance to harness the ability of using computational design to predict and optimize a synthetic genome before attempting its synthesis. The methodology to computationally design a genome is based on an optimization that computationally mimics genome evolution. The biggest bottleneck lies on the use of an appropriate fitness function. This fitness function, usually cell growth, relies on the ability to quantitatively model the biochemical networks of the cell at the genome scale using parameters inferred from high-throughput data. Computational methods integrating such models in a common multilayer design platform can be used to automatically engineer synthetic genomes under physiological specifications. We describe the current state-of-the-art on automated methods for engineering or re-engineering synthetic genomes. We restrict ourselves to global models of metabolism, transcription and DNA structure. Although we are still far from the de novo computational genome design, it is important to collect all relevant work towards this goal. Finally, we discuss future perspectives about the practicability of an automated methodology for such computational design of synthetic genomes. PMID- 19562113 TI - Immobilisation and encapsulation of functional protein-inorganic constructs. AB - Self-assembly methods for the immobilisation or encapsulation of the positively charged redox protein, cytochrome c (cyt c), in layered organoclays or silica nanoparticles, respectively, are described and contrasted. Protein-polymer organoclay nanocomposites are produced by spontaneous restacking of delaminated aminopropyl-functionalised magnesium phyllosilicate sheets in the presence of an aqueous solution of poly(sodium 4-styrene sulfonate) (PSS) and cyt c. In contrast, single molecules of cyt c are encapsulated in silica nanoparticles by sol-gel reactions at the oil-water interface of microemulsion water droplets. In both cases, the protein molecules remain structurally intact after entrapment, are accessible to small molecule redox agents, exhibit excellent peroxidase activity in the presence of hydrogen peroxide, and show enhanced stability and catalytic properties under adverse conditions of pH. The ability to prepare functional protein-inorganic conjugates in general could significantly extend the technological scope of biological products and processes, and should therefore be an important adjunct in the translation of synthetic biology to real-life applications. PMID- 19562114 TI - Role of a salt bridge in the model protein crambin explored by chemical protein synthesis: X-ray structure of a unique protein analogue, [V15A]crambin-alpha carboxamide. AB - We have used total chemical synthesis to prepare [V15A]crambin-alpha-carboxamide, a unique protein analogue that eliminates a salt bridge between the delta guanidinium of the Arg(10) side chain and the alpha-carboxylate of Asn(46) at the C-terminus of the polypeptide chain. This salt bridge is thought to be important for the folding and stability of the crambin protein molecule. Folding, with concomitant disulfide bond formation, of the fully reduced [V15A]crambin-alpha carboxamide polypeptide was less efficient than folding/disulfide formation for the [V15A]crambin polypeptide under a standard set of conditions. To probe the origin of this less efficient folding/disulfide bond formation, we separately crystallized purified synthetic [V15A]crambin-alpha-carboxamide and chemically synthesized [V15A]crambin and solved their X-ray structures. The crystal structure of [V15A]crambin-alpha-carboxamide showed that elimination of the Arg(10)-Asn(46) salt bridge caused disorder of the C-terminal region of the polypeptide chain and affected the overall 'tightness' of the structure of the protein molecule. These studies, enabled by chemical protein synthesis, strongly suggest that in native crambin the Arg(10)-Asn(46) salt bridge contributes to efficient formation of correct disulfide bonds and also to the well-ordered structure of the protein molecule. PMID- 19562115 TI - A synthetic metabolite-based mammalian inter-cell signaling system. AB - Functionally well-characterized modular transcription units represent the genetic repertoire for the design of synthetic gene networks operating inside individual mammalian cells. Interconnection of specialized cells to multicellular assemblies that could execute complex computational functions requires synthetic signaling systems, which process and synchronize metabolic information between mammalian cells. In this study we have designed a metabolite-controlled inter-cellular signaling device consisting of a human sender cell line stably engineered for constitutive expression of the human liver-type arginase and a transgenic receiver cell line harboring a synthetic circuit, which produced a human glycoprotein in response to L-arginine levels in the culture medium. Quantitative characterization of the system components enabled precise prediction of l arginine degradation and product gene expression kinetics and showed that two independent transgenic cell lines could functionally inter-operate to form a metabolite-controlled device which is able to precisely time desired target gene expression. Synthetic gene circuits modulating the transfer of metabolic information from a sender to a receiver cell line may enable the design of synthetic hormone systems supporting communication across multicellular assemblies. PMID- 19562116 TI - Expanded chemical diversity sampling through whole protein evolution. AB - A directed evolution method has been developed that allows random substitution of a contiguous trinucleotide sequence for TAG throughout a target gene for use in conjunction with an expanded genetic code. Using TEM-1 beta-lactamase and enhanced green fluorescent protein as targets, protein variants were identified whose functional phenotype was rescued in vivo when co-expressed with orthogonal tRNA-aminoacyl-tRNA synthase pairs that insert p-iodophenylalanine in response to UAG. Sequencing of the selected clones that retained the target protein function revealed that >90% of the variants contained in-frame TAG codons distributed throughout the target gene. Such an approach will allow broader sampling of new chemical diversity by proteins, so opening new avenues for studying biological systems and for adapting proteins for biotechnological applications. A common set of reagents allows the method to be used on different protein systems and in combination with an array of different unnatural amino acids, so helping to reveal the true potential for engineering proteins through expanded chemical diversity sampling. PMID- 19562121 TI - Ultrasound-assisted synthesis of carbon materials. AB - In this Perspective, the use of ultrasound in the synthesis and modification of carbon materials is surveyed. Ultrasound is a common laboratory tool used to nebulize solutions into fine mists, emulsify mixtures, and drive chemical reactions. Given society's continued use of carbon materials (e.g., carbon black and activated carbon) as pigments, adsorbents, and composite components as well as the exciting new applications being explored for carbon nanotubes, graphene, and meso- and macroporous carbons, the use of ultrasound in the synthesis and modification of carbon materials is of general interest. Here, carbon materials prepared by both ultrasonic spray pyrolysis and high intensity ultrasound will be discussed, with the properties and applications enabled by their preparation highlighted within the individual examples. This article is concluded with some personal perspectives on the directions toward which future research may be aimed. PMID- 19562122 TI - Capacitance of KOH activated carbide-derived carbons. AB - Chemical activation of CDC leads to capacitance values in an organic electrolyte as high as 180 F/g, 30% larger than those of as-produced samples, due to the surface area and microporosity development occurring as a consequence of the activation. PMID- 19562123 TI - Size effect on UV-Vis absorption properties of colloidal C(60) particles in water. AB - An obvious size effect of colloidal C(60) particles on UV-Vis absorption spectra was found after size fractionation of colloidal C(60) particles in water using the Field Flow Fractionation method. PMID- 19562124 TI - Synthesis of tripodal anchor units bearing selenium functional groups and their adsorption behaviour on gold. AB - The synthesis of new selenium-functionalized tripodal anchor units composed by a tetraphenylmethane core with three selenocyanate or selenol arms has been successfully accomplished and CV, XPS and UPS measurements of their monolayers on a gold surface were investigated. PMID- 19562125 TI - Oxidation of glycerol using gold-palladium alloy-supported nanocrystals. AB - The use of bio-renewable resources for the generation of materials and chemicals continues to attract significant research attention. Glycerol, a by-product from biodiesel manufacture, is a highly functionalised renewable raw material, and in this paper the oxidation of glycerol in the presence of base using supported gold, palladium and gold-palladium alloys is described and discussed. Two supports, TiO(2) and carbon, and two preparation methods, wet impregnation and sol-immobilisation, are compared and contrasted. For the monometallic catalysts prepared by impregnation similar activities are observed for Au and Pd, but the carbon-supported monometallic catalysts are more active than those on TiO(2). Glycerate is the major product and lesser amounts of tartronate, glycolate, oxalate and formate are observed, suggesting a sequential oxidation pathway. Combining the gold and palladium as supported alloy nanocrystals leads to a significant enhancement in catalyst activity and the TiO(2)-supported catalysts are significantly more active for the impregnated catalysts. The use of a sol immobilisation preparation method as compared to impregnation leads to the highest activity alloy catalysts and the origins of these activity trends are discussed. PMID- 19562126 TI - On the concept of ionicity in ionic liquids. AB - Ionic liquids are liquids comprised totally of ions. However, not all of the ions present appear to be available to participate in conduction processes, to a degree that is dependent on the nature of the ionic liquid and its structure. There is much interest in quantifying and understanding this 'degree of ionicity' phenomenon. In this paper we present transport data for a range of ionic liquids and evaluate the data firstly in terms of the Walden plot as an approximate and readily accessible approach to estimating ionicity. An adjusted Walden plot that makes explicit allowance for differences in ion sizes is shown to be an improvement to this approach for the series of ionic liquids described. In some cases, where diffusion measurements are possible, it is feasible to directly quantify ionicity via the Nernst-Einstein equation, confirming the validity of the adjusted Walden plot approach. Some of the ionic liquids studied exhibit ionicity values very close to ideal; this is discussed in terms of a model of a highly associated liquid in which the ion correlations have similar impact on both the diffusive and conductive motions. Ionicity, as defined, is thus a useful measure of adherence to the Nernst-Einstein equation, but is not necessarily a measure of ion availability in the chemical sense. PMID- 19562127 TI - Insights into affinity and specificity in the complexes of alpha-lytic protease and its inhibitor proteins: binding free energy from molecular dynamics simulation. AB - We report the binding free energy calculation and its decomposition for the complexes of alpha-lytic protease and its protein inhibitors using molecular dynamics simulation. Standard mechanism serine protease inhibitors eglin C and OMTKY3 are known to have strong binding affinity for many serine proteases. Their binding loops have significant similarities, including a common P1 Leu as the main anchor in the binding interface. However, recent experiments demonstrate that the two inhibitors have vastly different affinity towards alpha-lytic protease (ALP), a bacterial serine protease. OMTKY3 inhibits the enzyme much more weakly (by approximately 10(6) times) than eglin C. Moreover, a variant of OMTKY3 with five mutations, OMTKY3M, has been shown to inhibit 10(4) times more strongly than the wild-type inhibitor. The underlying mechanisms for the unusually large difference in binding affinities and the effect of mutation are not well understood. Here we use molecular dynamics simulation with molecular mechanics Poisson Boltzmann/surface area method (MM-PB/SA) to investigate quantitatively the binding specificity. The calculated absolute binding free energies correctly differentiate the thermodynamic stabilities of these protein complexes, but the magnitudes of the binding affinities are systematically overestimated. Analysis of the binding free energy components provides insights into the molecular mechanism of binding specificity. The large DeltaDeltaG(bind) between eglin C and wild type OMTKY3 towards ALP is mainly attributable to the stronger nonpolar interactions in the ALP-eglin C complex, arising from a higher degree of structural complementarity. Here the electrostatic interaction contributes to a lesser extent. The enhanced inhibition in the penta-mutant OMTKY3M over its wild type is entirely due to an overall improvement in the solvent-mediated electrostatic interactions in the ALP-OMTKY3M complex. The results suggest that for these protein-complexes and similar enzyme-inhibitor systems (1) the binding is driven by nonpolar interactions, opposed by overall electrostatic and solute entropy contributions; (2) binding specificity can be tuned by improving the complementarity in electrostatics between two associating proteins. Binding free energy decomposition into contributions from individual protein residues provides additional detailed information on the structural determinants and subtle conformational changes responsible for the binding specificity. PMID- 19562128 TI - Ar, CCl(4) and C(6)H(6) adsorption outside and inside of the bundles of multi walled carbon nanotubes-simulation study. AB - This is the first paper reporting the results of systematic study of the adsorption of Ar, C(6)H(6) and CCl(4) on the bundles of closed and opened multi walled carbon nanotubes. Using grand canonical Monte Carlo (GCMC) and molecular dynamics (MD) simulations, we also study the effect of the introducing defects in the external and internal walls of osculating and separated nanotubes on Ar diffusion and on adsorption of all three adsorbates. The Ar diffusion coefficients obtained are very sensitive to the presence of defects. Simulated isotherms are discussed to show the relation between the shapes of the high resolution alpha(s)-plots and the mechanisms of adsorption. From obtained data, as well as from geometric considerations, from the VEGA ZZ package, and from simulations (ASA), the values of surface areas of all nanotubes are calculated and compared with those obtained using the most popular adsorption methods (BET, alpha(s) and the A,B,C-points). We show that the adsorption value for the C-point of the isotherm should be taken for the calculation of the specific surface area of carbon nanotubes to obtain a value which approaches the absolute geometric surface area. A fully packed monolayer is not created at the A-, B- or C-points of the isotherm; however, the number of molecules adsorbed at the latter point is closest to the number of molecules in the monolayer as calculated via the ASA method, the VEGA ZZ package or from geometric considerations. PMID- 19562129 TI - Dynamics of guest molecules in PHTP inclusion compounds as probed by solid-state NMR and fluorescence spectroscopy. AB - Partially deuterated 1,4-distyrylbenzene () is included into the pseudohexagonal nanochannels of perhydrotriphenylene (PHTP). The overall and intramolecular mobility of is investigated over a wide temperature range by (13)C, (2)H NMR as well as fluorescence spectroscopy. Simulations of the (2)H NMR spectral shapes reveal an overall wobble motion of in the channels with an amplitude of about 4 degrees at T = 220 K and 10 degrees at T = 410 K. Above T = 320 K the wobble motion is superimposed by localized 180 degrees flips of the terminal phenyl rings with a frequency of 10(6) Hz at T = 340 K. The activation energies of both types of motions are around 40 kJ mol(-1) which imply a strong sterical hindrance by the surrounding PHTP channels. The experimental vibrational structure of the fluorescence excitation spectra of is analyzed in terms of small amplitude ring torsional motions, which provide information about the spatial constraints on by the surrounding PHTP host matrix. Combining the results from NMR and fluorescence spectroscopy as well as of time-dependent density functional calculations yields the complete potential surfaces of the phenyl ring torsions. These results, which suggest that intramolecular mobility of is only reduced but not completely suppressed by the matrix, are corroborated by MD simulations. Unrealistically high potential barriers for phenyl ring flips are obtained from MD simulations using rigid PHTP matrices which demonstrate the importance of large amplitude motions of the PHTP host lattice for the mobility of the guest molecules. PMID- 19562130 TI - Characterising the phase behaviour of stearic acid and its triethanolamine soap and acid-soap by infrared spectroscopy. AB - The behaviour of stearic acid neutralised by triethanolamine to form soap and its acid-soap has been examined by infrared spectroscopy. It was found that not only could the neutralisation behaviour be characterised, but the thermotropic behaviour could also be followed. The neutralisation confirmed the formation of a fixed stoichiometeric ratio, 2 : 1, acid-soap. When following the thermotropic behaviour the break up of the acid-soap could be followed along with various disordering and melting transitions of the alkyl chain tail. This allowed all the thermal transitions that have been observed to be characterised in terms of the type of molecular rearrangement that was occurring and also the transition temperature at which they occurred. This allowed the binary phase diagram to be plotted and understood for this system. This is the first time IR has been used to measure a whole phase diagram of this type. The nature of the acid-soap complex itself was also characterised, with very short hydrogen bonds being present as well as a free, non-hydrogen bonded, hydroxyl group. PMID- 19562131 TI - Ring-opening reaction of a trifluorinated indolylfulgide: mode-specific photochemistry after pre-excitation. AB - The ring-opening reaction of a trifluorinated indolylfulgide has been studied as a function of temperature and optical pre-excitation where it was found that reaction times decreased as temperature increased from 10.3 ps at 12 degrees C to 7.6 ps at 60 degrees C. Simultaneously, the quantum yields for the ring-opening reaction grew from 3.1% (12 degrees C) to 5.0% (60 degrees C). When the reaction was started from a non-equilibrium state generated by a directly preceding ring closure process, the ring-opening reaction became faster and the quantum efficiency increased by more than a factor of three. Analysis of the experimental results points to mode-specific photochemistry in that the promoting, photochemically active modes of the photoreaction are efficiently excited by the directly preceding ring-closure reaction. PMID- 19562132 TI - Intermolecular forces in lipid monolayers. Two-dimensional virial coefficients for pentadecanoic acid from micromanometry on spread monolayers at the air/water interface. AB - The lateral intermolecular forces between surfactant or lipid molecules in monolayers at interfaces are fundamental to understanding the phenomena of surface activity and the interactions of lipids in two-dimensional structures such as smectic phases and biomembranes. The classical approach to these forces is via the two-dimensional virial coefficients, which requires precise micromanometry on monolayer isotherms in the dilute gaseous region. Low pressure isotherms out to high surface areas in the two-dimensional gas range have been measured at 15, 25 and 30 degrees C for insoluble monolayers of n-pentadecanoic acid spread at the interface between water-vapour saturated air and a dilute aqueous solution of HCl. The data allow estimates of virial coefficients up to the third term. The second virial coefficients are compared with those predicted from a statistical mechanical model for monolayers of n-alkylcarboxylic acids treated as side-by-side parallel chains extended at the surface with the carboxyl head groups shielded in the water phase. The two sets coincide at approximately 26 degrees C, but the experimental estimates show a much larger dependence on temperature than the model predicts. Chain conformation effects, head group interactions and surface field polarization are discussed as possible temperature dependent contributions to the lateral potentials of mean force. PMID- 19562133 TI - Thermal expansivity of amyloid beta(16-22) peptides and their aggregates in water. AB - Temperature dependence of the volumetric and structural properties of Abeta(16 22) peptides (wild type and pathogenic forms) and their aggregates in water was studied by simulations. The intrinsic thermal expansion coefficient alpha(p) of peptides was evaluated by taking into account the difference between the volumetric properties of hydration and bulk water. Single peptides show mainly positive values of alpha(p) that correlates with the increasing number of intrapeptide hydrogen bonds upon heating. Negative values of alpha(p) found for large peptide aggregates may be attributed to the shrinking of voids inside aggregates with increasing temperature or to their rubber-like elasticity. The peptide surface exposed to water becomes more hydrophobic with increasing aggregate size that appears in decreasing density of hydration water and evidences a hydrophilic character of aggregation. PMID- 19562134 TI - Development of a new analysis method evaluating adsorption energies for the respective ion-exchanged sites on alkali-metal ion-exchanged ZSM-5 utilizing CO as a probe molecule: IR-spectroscopic and calorimetric studies combined with a DFT method. AB - For alkali-metal ion-exchanged ZSM-5 zeolites (MZSM-5; M: Li, Na, K, Rb, Cs) the analysis of ion-exchangeable sites was performed by means of a combined method based on IR spectroscopic and calorimetric measurements using CO as the probe molecule. The heat of adsorption of CO was found to be correlated with an IR frequency of stretching vibration of C-O in the adsorbed species. It was revealed that there exists at least two types of sites capable of ion-exchanging; for the lithium ion-exchanged ZSM-5 (LiZSM-5) CO adsorption on each type of site is evaluated to give a set of IR bands and heats of adsorption, 2195 cm(-1) and 49 kJ mol(-1), 2185 cm(-1) and 39 kJ mol(-1) with the aid of the newly developed method utilizing the data obtained from a combined microcalorimetric and IR spectroscopic study. Such types of data were also obtained for Na- and K-ion exchanged ZSM-5 samples. Furthermore, a linear relationship between the differential heat of adsorption (q(diff)) evaluated and the shift of wavenumber of the C-O stretching vibration from that of a gaseous CO molecule (Deltanu) was established for the systems of MZSM-5-CO, and the bonding nature of the CO molecule with each site can be explained in terms of the electrostatic force. The model of each adsorption site was also examined by the quantum calculation method (density functional theory: DFT). The trends obtained from the experimental data may be substantially supported by the calculation method even adopting a model as simple as the ZSM-5-type zeolite: the composition of MAlSi(4)O(4)H(12). PMID- 19562135 TI - Substituent effect on the interaction of aromatic primary amines and diamines with supercritical CO(2) from infrared spectroscopy and quantum calculations. AB - The nature and the strength of the interactions occurring between aromatic primary amines and CO(2) have been investigated by combining infrared spectroscopy with molecular modelling. A series of infrared absorption experiments on various aromatic mono- and diamines in supercritical CO(2) have been performed at constant temperature (T = 40 degrees C) for various CO(2) pressures varying from 6 to 30 MPa. Then, we carried out a theoretical analysis based on quantum calculations using both density functional (B3LYP) and ab initio (MP2) computational methods. Whatever the amine considered, CO(2) is found to be on average above the nitrogen atom of the NH(2) group for which the donating lone pair interacts with the carbon atom of CO(2). Several types of interactions have been identified, namely, electron donor-acceptor (EDA), hydrogen bonds as well as dispersion forces. Contrary to aliphatic amines, the dispersion interaction is significant in the aromatic amine-CO(2) complexes because of the presence of the aromatic ring. The substituents are found to influence the stability and structure of the amine-CO(2) complexes, directly by electrostatic and steric effects of the substituent, and indirectly through the change in the partial charge on the nitrogen atom. Finally, a good correlation has been put in evidence between the partial charge on the nitrogen atom and the EDA interaction occurring between the aromatic amines and CO(2). PMID- 19562136 TI - Photoluminescence of CdTe nanocrystals modulated by methylene blue and DNA. A label-free luminescent signaling nanohybrid platform. AB - A nanohybrid consisting of water-soluble thioglycolic acid (TGA)-capped CdTe nanocrystals (NCs) and methylene blue (MB) was designed as a label-free luminescent signaling platform for DNA. This sensing system was identified to operate under the photoinduced electron transfer (PET) mechanism in which MB is the electron acceptor and the binding site for the designated target molecule DNA. We showed that MB bound with TGA-capped CdTe NCs via strong electrostatic interactions resulted in an efficient quenching of the photoluminescence (PL) of NCs. Steady-state and time-resolved PL, and electron paramagnetic resonance (EPR) experiments established the quenching pathway of PET from the conduction band (CB) of NCs to the ground state of MB. In the presence of the target molecule DNA, the MB-quenched PL of NCs could be reversibly restored by double-stranded DNA as the PET pathway is blocked when MB is taken away from the NCs surface due to its intercalation into, and electrostatic interaction with, DNA. The platform was successfully applied for sensing DNA and signaling DNA hybridization by switching the PET process. Such a nanohybrid represents a robust PET luminescent nanosensor that is, in principle, applicable for other species by employing suitable electron acceptors as binding sites. PMID- 19562137 TI - Thermodynamics and structure of inclusion compounds of tauro- and glyco conjugated bile salts and beta-cyclodextrin. AB - The interaction between natural beta-cyclodextrin and bile salts common in rat, dog and man, taurocholate, tauro-beta-muricholate, taurodeoxycholate, taurochenodeoxycholate, glycocholate, glycodeoxycholate and glycochenodeoxycholate, was studied using isothermal titration calorimetry, and the structural differences in the interaction were investigated by (1)H-ROESY NMR and molecular modeling. The beta-cyclodextrin was selected based upon its frequent use in preformulation and drug formulation as oral excipients for the solubilization of drug substances with low aqueous solubility. All the investigated bile salts possessed affinity for the cyclodextrin, though with large variations in the stability constants. The variations in the enthalpic and entropic contributions to the overall Gibbs free energy and consequently the stability constants revealed differences in the binding mode between the investigated bile salts, i.e. the bile salts with a hydroxyl group on C12 interacted differently from the bile salts without this hydroxyl group. These observations were supported by both (1)H-ROESY NMR and molecular modeling, which suggested binding on the D-ring in the steroid structure for the former and on the C-ring for the latter bile salts. PMID- 19562138 TI - Phase diagram of Ag-Pd bimetallic nanoclusters by molecular dynamics simulations: solid-to-liquid transition and size-dependent behavior. AB - This report on the solid-to-liquid transition region of an Ag-Pd bimetallic nanocluster is based on a constant energy microcanonical ensemble molecular dynamics simulation combined with a collision method. By varying the size and composition of an Ag-Pd bimetallic cluster, we obtained a complete solid-solution type of binary phase diagram of the Ag-Pd system. Irrespective of the size and composition of the cluster, the melting temperature of Ag-Pd bimetallic clusters is lower than that of the bulk state and rises as the cluster size and the Pd composition increase. Additionally, the slope of the phase boundaries (even though not exactly linear) is lowered when the cluster size is reduced on account of the complex relations of the surface tension, the bulk melting temperature, and the heat of fusion. The melting of the cluster initially starts at the surface layer. The initiation and propagation of a five-fold icosahedron symmetry is related to the sequential melting of the cluster. PMID- 19562139 TI - Synergistic interaction between gold nanoparticles and nickel phthalocyanine in layer-by-layer (LbL) films: evidence of constitutional dynamic chemistry (CDC). AB - The concept of constitutional dynamic chemistry (CDC) based on the control of non covalent interactions in supramolecular structures is promising for having a large impact on nanoscience and nanotechnology if adequate nanoscale manipulation methods are used. In this study, we demonstrate that the layer-by-layer (LbL) technique may be used to produce electroactive electrodes with ITO coated by tetrasulfonated nickel phthalocyanine (NiTsPc) alternated with poly(allylamine hydrochloride) (PAH) incorporating gold nanoparticles (AuNP), in which synergy has been achieved in the interaction between the nanoparticles and NiTsPc. The catalytic activity toward hydrogen peroxide (H(2)O(2)) in multilayer films was investigated using cyclic voltammetry, where oxidation of H(2)O(2) led to increased currents in the PAH-AuNP/NiTsPc films for the electrochemical processes associated with the phthalocyanine ring and nickel at 0.52 and 0.81 V vs. SCE, respectively, while for PAH/NiTsPc films (without AuNP) only the first redox process was affected. In control experiments we found out that the catalytic activity was not solely due to the presence of AuNP, but rather to the nanoparticles inducing NiTsPc supramolecular structures that favored access to their redox sites, thus yielding strong charge transfer. The combined effects of NiTsPc and AuNP, which could only be observed in nanostructured LbL films, point to another avenue to pursue within the CDC paradigm. PMID- 19562140 TI - The role of water in silicate oligomerization reaction. AB - The silicate oligomerization reaction is key to sol-gel chemistry and zeolite synthesis. Numerous experimental and theoretical studies have been devoted to investigating the physical chemistry of silicate oligomers in the prenucleation stage of siliceous zeolite formation. Most of the previous quantum chemical computational work has used gas phase models or continuous solvent models for silica oligomerization. Here we apply Car-Parrinello molecular dynamics simulations with explicit inclusion of water molecules to investigate the reaction pathway for the anionic bond formation of siliceous oligomers. The rates of SiO-Si bond formation of linear or ring containing silicate oligomers become substantially enhanced, compared to gas phase results. The formation of 3-ring oligomer is more favorable than the formation of higher branched and ring silica oligomers. PMID- 19562141 TI - Theoretical study of proton encircling modes in proton sponges with tetraamido/diamino quaternized macrocycles: the role of pi-conjugated and aliphatic bridges. AB - Theoretical studies of the proton encapsulation within a series of tetraamido/diamino quaternized macrocycles consisting of -NH-CO- functionalities linked by different pi-conjugated (phenyl, pyridine, furan, thiophene, butadienyl, and ethylene) or aliphatic (propyl and butyl) bridges are carried out. In macrocycles containing pi-conjugated spacers, the protons are held by asymmetrical hydrogen bonds O-HO right harpoon over left harpoon OH-O with very small barriers for the proton transfer. In the case of macrocycles with aliphatic spacers, a symmetrical hydrogen bond (OHO) is formed in the propyl analogue, in contrast to the asymmetrical hydrogen bond in the butyl analogue. The energy based fragmentation quantum chemistry method is employed to calculate the hydrogen bond energies of macrocycles. Taking advantage of the fragment-based calculations, contributions from individual bridges to the strength of hydrogen bonds are also revealed. A qualitative agreement is shown between fragmentation MP2/6-31G(d,p) calculation results, and the hydrogen bond energies estimated from the empirical equation on the basis of O-H stretching frequency. The close correlation between the hydrogen bond energy with electrostatic interactions within the hydrogen bond framework, and the proton affinity of macrocycles, indicate that the intramolecular hydrogen bonds are mostly electrostatic. The calculations show that the modifications of the bridges linking the -NH-CO- functionalities and hence the OO distance between the proton acceptor and donor determine the proton binding modes. PMID- 19562142 TI - Dielectric and calorimetric investigation of an unusual two-component plastic crystal: cyclohexanol-neopentylglycol. AB - In the present article, investigations of an unusual two-component (H-) bonded pair, i.e. the cyclohexanol-neopentylglycol system, are reported. The phase I of cyclohexanol (CHXOL) forms a continuous solid solution with the phase I of neopentylglycol (NPGOL). This binary solid solution (S(I)) has been investigated at low temperatures and several concentrations, by means of dielectric spectroscopy and differential scanning calorimetry (DSC). Depending upon the concentration, this phase reveals a glass transition in the temperature range 150 180 K and a pronounced relaxation process identifiable with the so-called primary relaxation process, or alpha-process. The analysis of the various parameters obtained shows an isomorphic relationship between the face-centered cubic phases of both the pure components through a continuous change of parameters. In addition, two sub-T(g) processes (designated as beta-and gamma-) are found. The present observation suggests that the beta-process is probably a Johari-Goldstein relaxation process and the gamma-process is intramolecular in nature. The kinetic freezing of the various dielectric processes has been examined in relation to the T(g) found in the DSC experiments. PMID- 19562143 TI - A novel europium(III) complex with versatility in excitation ranging from infrared to ultraviolet. AB - One- and two-photon absorption properties of a novel europium(III) complex containing a hemicyanine cation are investigated in this paper; in which the sensitization wavelength of europium(III), induced by beta-diketonate ligands, is in the ultraviolet range, and the hemicyanine(aminostyrylpyridinium) cation extends the sensitization wavelength of Eu(III) to visible region. Furthermore, under 1.06 microm ultrashort pulse laser excitation, the complex exhibits effective europium(III) luminescence induced by the two-photon absorption of the hemicyanine cation. Therefore, this new europium(III) complex with excitation bands in the near infrared, visible and ultraviolet ranges. Since the 1.06-microm ultrafast laser is one of the most frequently used laser sources and 1.06 microm is an 'optical window' for cells and tissues, the results suggest that this kind of complex has promising applications in deep-penetrating and high quality bioimaging in vivo. PMID- 19562145 TI - Ion beam degradation analysis of poly(3-hexylthiophene) (P3HT): can cryo-FIB minimize irradiation damage? AB - In this study, to assess the influence of the temperature on the ion beam degradation, irradiation experiments on organic semiconductor materials were performed for both cryogenic and room temperature conditions. Thin P3HT films on silicon substrates were exposed to increasing ion doses in dual beam FIB. The degradation behaviour by means of a decrease in the C[double bond, length as m dash]C band which corresponds to a loss of conjugation was investigated by means of Raman spectroscopy. In addition, atomic force microscopy (AFM) and Kelvin probe force microscopy (KPFM) were used for a characterization of morphology and surface potential which provide information on temperature and ion dose dependent degradation behaviour. PMID- 19562144 TI - Improving the accuracy of low level quantum chemical calculation for absorption energies: the genetic algorithm and neural network approach. AB - The combination of genetic algorithm and back-propagation neural network correction approaches (GABP) has successfully improved the calculation accuracy of absorption energies. In this paper, the absorption energies of 160 organic molecules are corrected to test this method. Firstly, the GABP1 is introduced to determine the quantitative relationship between the experimental results and calculations obtained by using quantum chemical methods. After GABP1 correction, the root-mean-square (RMS) deviations of the calculated absorption energies reduce from 0.32, 0.95 and 0.46 eV to 0.14, 0.19 and 0.18 eV for B3LYP/6-31G(d), B3LYP/STO-3G and ZINDO methods, respectively. The corrected results of B3LYP/6 31G(d)-GABP1 are in good agreement with experimental results. Then, the GABP2 is introduced to determine the quantitative relationship between the results of B3LYP/6-31G(d)-GABP1 method and calculations of the low accuracy methods (B3LYP/STO-3G and ZINDO). After GABP2 correction, the RMS deviations of the calculated absorption energies reduce to 0.20 and 0.19 eV for B3LYP/STO-3G and ZINDO methods, respectively. The results show that the RMS deviations after GABP1 and GABP2 correction are similar for B3LYP/STO-3G and ZINDO methods. Thus, the B3LYP/6-31G(d)-GABP1 is a better method to predict absorption energies and can be used as the approximation of experimental results where the experimental results are unknown or uncertain by experimental method. This method may be used for predicting absorption energies of larger organic molecules that are unavailable by experimental methods and by high-accuracy theoretical methods with larger basis sets. The performance of this method was demonstrated by application to the absorption energy of the aldehyde carbazole precursor. PMID- 19562146 TI - Study of the Beckmann rearrangement of acetophenone oxime over porous solids by means of solid state NMR spectroscopy. AB - The Beckmann rearrangement of acetophenone oxime using zeolite H-beta and silicalite-N as catalysts has been investigated by means of (15)N and (13)C solid state NMR spectroscopy in combination with theoretical calculations. The results obtained show that the oxime is N-protonated at room temperature on the acid sites of zeolite H-beta. At reaction temperatures of 423 K or above, the two isomeric amides, acetanilide and N-methyl benzamide (NMB) are formed, and interact with the Bronsted acid sites of zeolite H-beta through hydrogen bonds. The presence of residual water hydrolyzes the two amides, while larger amounts inhibit the formation of NMB and cause the total hydrolysis of the acetanilide. Over siliceous zeolite silicalite-N, containing silanol nests as active sites, the oxime is adsorbed through hydrogen bonds and only acetanilide is formed at reaction temperatures of 423 K or above. In the presence of water, the reaction starts at 473 K, still being very selective up to 573 K, and the amide is partially hydrolyzed only above this temperature . PMID- 19562147 TI - Solvent-free oxidation of benzyl alcohol using Au-Pd catalysts prepared by sol immobilisation. AB - We report the preparation of Au-Pd nanocrystalline catalysts supported on TiO(2) and carbon prepared via a sol-immobilisation technique using three different preparation strategies; namely, simultaneous formation of the sols for both metals or initial formation of a seed sol of one of the metals followed by a separate step in which a coating sol of the second metal is added. The catalysts have been structurally characterised using a combination of transmission electron microscopy and X-ray photoelectron spectroscopy. The catalysts have been evaluated for the oxidation of benzyl alcohol under solvent-free conditions. The catalysts prepared using the sol immobilisation technique show higher activity when compared with catalysts prepared by impregnation, particularly as lower metal concentrations can be used. The Au-Pd catalysts were all more active than the corresponding monometallic supported Au or Pd catalysts. For 1 wt% Au Pd/TiO(2) the order of metal addition in the preparation was not observed to be significant with respect to selectivity or activity. However, the 1 wt% Au Pd/carbon catalysts are more active but less selective to benzaldehyde than the TiO(2)-supported catalysts when compared at iso-conversion. Furthermore, for the carbon-supported catalyst the order of metal addition has a very marked affect on activity. The carbon-supported catalysts are also more significantly affected by heat treatment, e.g. calcination at 400 degrees C leads to the activity being decreased by an order of magnitude, whereas the TiO(2)-supported catalysts show a 50% decrease in activity. Toluene is observed as a by-product of the reaction and conditions have been identified that minimise its formation. It is proposed that toluene and benzaldehyde are formed by competing parallel reactions of the initial benzyl intermediate via an adsorbed benzylidene species that can either be hydrogenated or oxidised. Hence, conditions that maximise the availability of oxygen on the catalyst surface favour the synthesis of benzaldehyde. PMID- 19562148 TI - H/D isotope effects on NMR chemical shifts of nuclei involved in a hydrogen bridge of hydrogen isocyanide complexes with fluoride anion. AB - (1)H, (2)H, (19)F and (15)N NMR spectra of a strongly hydrogen-bonded anionic cluster, CNHF(-), as an ion pair with a tetrabutylammonium cation dissolved in CDF(3)-CDF(2)Cl mixture were recorded in the slow exchange regime at temperatures down to 110 K. The fine structure due to spin-spin coupling of all nuclei involved in the hydrogen bridge was resolved. H/D isotope effects on the chemical shifts were measured. The results were compared with those obtained earlier for a similar anion, FHF(-), and interpreted via ab initio calculations of magnetic shielding as functions of internal vibrational coordinates, namely an anti symmetric proton stretching and a doubly-degenerate bending. The values of primary and secondary isotope effects on NMR chemical shifts were estimated using a power expansion of the shielding surface as a function of vibrational coordinates. A positive primary isotope effect was explained as a result of the decrease of the hydron stretching amplitude upon deuteration. We show that the proton shielding surface has a minimum close to the equilibrium geometry of the CNHF(-) anion, leading to the positive primary H/D isotope effect in a rather asymmetric hydrogen bond. We conclude that caution should be used when making geometric estimations on the basis of NMR data, since the shapes of the shielding functions of the internal vibrational coordinates can be rather exclusive for each complex. PMID- 19562149 TI - Chirality-dependent hydrogen bond direction in jet-cooled (S)-1,2,3,4-tetrahydro 3-isoquinoline methanol (THIQM): IR-ion dip vibrational spectroscopy of the neutral and the ion. AB - The structural modifications of (S)-1,2,3,4-tetrahydro-3-isoquinoline methanol (THIQM) upon ionisation have been investigated in jet-cooled conditions, by means of laser-induced fluorescence, REMPI, and IR-UV ion-dip spectroscopy of the neutral ground state and the ion. These results combined with ab initio calculations, support the presence in the jet of two low-energy conformers of THIQM. In the most stable Conformer I, the CH(2)OH substituent acts as a hydrogen bond donor to the nitrogen lone pair in the equatorial position. In this case, the nitrogen atom is in (S) configuration. Conformer II shows the opposite NHO hydrogen bond from the hydrogen atom in the equatorial position of nitrogen to the OH group. In this case, the nitrogen atom is in (R) configuration. This chirality dependence of the hydrogen bond direction is lost upon ionisation. While ionisation of Conformer II reinforces the NHO hydrogen bond, ionisation of Conformer I induces its isomerisation to the same ion as Conformer II, i.e. a change in hydrogen bond direction. PMID- 19562150 TI - Dynamic solvent effects on the thermal isomerization of zinc dithizonate. AB - The pressure dependence of the rate constant for the thermal isomerization of zinc dithizonate was measured at various temperatures in highly viscous solvents, i.e., glycerol triacetate and 2,4-dicyclohexyl-2-methylpentane, as well as in their chemically similar nonviscous solvents, such as methyl acetate and methylcyclohexane. In the nonviscous solvents, the reaction was slightly retarded upon external pressure suggesting the validity of the transition state theory. On the other hand, in the viscous solvents, the reaction, even at ambient pressure, was strongly retarded by an increase in external pressure suggesting the slow thermal fluctuation of the solvent molecules upon external pressure in highly viscous systems. This made it possible to observe the dynamic solvent effects on the isomerization of zinc dithizonate in viscous solvents. The viscosity dependence of the rate constants was rationalized by the two-dimensional reaction coordinate model. PMID- 19562151 TI - Theoretical prediction of the solubility of fluorinated C(60). AB - Although extensive theoretical and experimental research has been conducted on fluorinated fullerenes, little detailed information exists on their solubility in different solvents. However, this solubility is crucial for their processability and possible application. In this work, we predict the solubility of fluorinated C(60) in various polar and non-polar solvents, based on a correlation between experimentally measured solubilities for C(60) from the literature and theoretically predicted solubilisation energies. These solubilisation energies are predicted using the polarizable continuum model (PCM) at the ab initio Hartree-Fock 6-31+G* level of theory. In particular, the solubilities are predicted for C(60)F(2)(n) (n = 1-10) isomers, part of the addition route to saturnene C(60)F(20). An increasing solubility is found for more polar solvents with higher degree of fluorination. With these results, we can determine the minimal fluorination degree necessary for possible solubilisation in a given solvent, and offer new perspectives to separate and purify species with different degrees of fluorination. PMID- 19562152 TI - Electromechanical actuation of macroscopic carbon nanotube structures: mats and aligned ribbons. AB - The electromechanical actuation of macroscopic carbon nanotube (CNT) structures, including single and multi walled CNT mats and aligned ribbons, is analyzed and compared. From experimental evidence, actuation due to quantum chemical and electrostatic effects can be distinguished. Their respective contribution to the total actuation depends on two key parameters, namely Young's modulus and charge density. While mechanical energy densities of actuated structures are appreciable, electromechanical conversion efficiencies are found to be impractically low. PMID- 19562153 TI - Electron transfer from A to A(1) in Photosystem I from Chlamydomonas reinhardtii occurs in both the A and B branch with 25-30-ps lifetime. AB - We have recorded transient absorption kinetics at 390 nm with picosecond resolution in order to observe electron transfer from the reduced primary acceptor, A, to the secondary acceptor, A(1), in wild type and mutated Photosystem I from Chlamydomonas reinhardtii. In the mutants, the methionine axial ligand to the primary electron acceptor in either the A- or B-branch of electron transfer cofactors, was replaced with histidine. Both of the mutations reduced the formation of a positive signal at 390 nm, characteristic of A to a level approximately half of that observed in wild type Photosystem I. It is concluded that in the mutated branch of Photosystem I, electron transfer from A to A(1) does not occur. The absorption kinetics resulting from subtraction of either of the mutants' traces from that of wild type is interpreted to reflect the kinetics of A- or B-side electron transfer from A to A(1) in the the wild type Photosystem I. Each of these traces could be fitted with a monoexpoenential decay characterized by the same amplitude and 25-30-ps lifetime. The almost identical effect of both mutations on A formation confirm a similar engagement of both the A- ad B-branches in electron transfer to A(1) in Photosystem I from C. reinhardtii. This observation is in contrast to the unidirectional electron transfer concluded from the studies on similar mutants of cyanobacterial Photosystem I.(1) Thus, this contribution provides further evidence for functional differences between these two model Photosystems. PMID- 19562154 TI - Preparation and characterization TiO(x)-Pt/C catalyst for hydrogen oxidation reaction. AB - The hydrogen oxidation reaction (HOR) was studied at the home made TiO(x)-Pt/C nanocatalysts in 0.5 mol dm(-3) HClO(4) at 25 degrees C. Pt/C catalyst was first synthesized by modified ethylene glycol method (EG) on commercially used carbon support (Vulcan XC-72). Then TiO(x)-Pt/C catalyst was prepared by the polyole method followed by TiO(x) post-deposition. The synthesized catalyst was characterized by XRD, TEM and EDX techniques. It was found that Pt/C catalyst nanoparticles were homogenously distributed over carbon support with the mean particle size of about 2.4 nm. The quite similar, homogenous distribution and particle size were obtained for Pt/C doped by TiO(x) catalyst which was the confirmation that TiO(x) post-deposition did not lead to significant growth of the Pt nanoparticles. The electrochemically active surface area of the catalyst was determined by using the cyclic voltammetry technique.The kinetics of hydrogen oxidation was investigated by the linear sweep voltammetry technique at the rotating disc electrode (RDE). The kinetic equations used for the analysis were derived considering the reversible or irreversible nature of the kinetics of the HOR. It was found that the hydrogen oxidation reaction for an investigated catalyst proceeded as an electrochemically reversible reaction. The values determined for the kinetic parameters-Tafel slope of 28 mV dec(-1) and exchange current density about 0.4 mA cm(-2)(Pt) are in good agreement with usually reported values for a hydrogen oxidation reaction with platinum catalysts in acid solutions. PMID- 19562160 TI - Synthetic applications of cationic iron and cobalt carbonyl complexes. AB - Metal carbonyl stabilized cationic species react with a wide range of nucleophiles under mild conditions, and have thus found many synthetic applications. In this Perspective, we describe the utility of iron carbonyl dienyl cations in solution and solid phase parallel synthesis, and in the development of a new synthetic route towards oseltamivir phosphate (Tamiflu). We also discuss the solid phase version of the Nicholas reaction, employing cobalt carbonyl stabilized propargylic cations, and giving access to substituted alkynes. PMID- 19562155 TI - A molecular wire incorporating a robust hexanuclear platinum cluster. AB - Reaction of [Pt(6)(CO)(4)(P(t)Bu(2))(4)Cl(2)] with excess HS(CH(2))(4)SH in Et(2)NH gave highly stable [Pt(6)(CO)(4)(P(t)Bu(2))(4){S(CH(2))(4)SH}(2)], which adsorbs unchanged onto gold surfaces. This permitted the fabrication and electrical characterisation of gold|molecule|gold junctions involving a well defined metal carbonyl cluster compound. PMID- 19562161 TI - A planar Cu2+ (S=1/2) kagome network pillared by 1,2-bis(4-pyridyl) ethane with interesting magnetic properties. AB - The reaction of Cu(ClO4)(2).6H2O and 1,2-bis(4-pyridyl)ethane (bpe) in basic aqueous solution yields a 3D compound, {[Cu3(CO3)2(bpe)3].2ClO4}n (1) through the atmospheric fixation of CO2 composing Cu(CO3) kagome layers pillared by the bpe linker that exhibit weak antiferromagnetic interaction in the kagome layer and interlayer ferromagnetic coupling at low temperature. PMID- 19562162 TI - Polynuclear titanocene complexes with antimony ligands: [(Cp2Ti)2(SbR2)2] (R=Et), [(Cp2Ti)3(SbR)3Sb] [R=2-(Me2NCH2)C6H4] and [(Cp2Ti)5(SbR)2Sb7] (R=Me3SiCH2). AB - The formation and crystal structure of [(Cp2Ti)2(SbR2)2] (1, R=Et), [(Cp2Ti)3(SbR)3Sb] [2, R=2-(Me2NCH2)C6H4], and [(Cp2Ti)5(SbR)2Sb7] (3, R=Me3SiCH2) is reported. PMID- 19562163 TI - The [1,2,3]triazolo[1,5-a]pyridine ring: a sensitive sensor for the electronic profile of phosphorus substituents. AB - The unique nature of the [1,2,3]triazolo[1,5-a]pyridine reveals without any external perturbation the electronic contribution of various substituents to the phosphorus atom in phosphines, based on the equilibrium of two possible ring chain isomers. PMID- 19562164 TI - A novel ruthenium(II) arene based intercalator with potent anticancer activity. AB - The new ruthenium(II) compound [(eta6-p-cymene)Ru(N insertion mark C)Cl] (HN insertion mark C=9-aminoacridine (9-HAA)) shows a potent in vitro anticancer activity. PMID- 19562165 TI - Para-hydrogen induced polarisation effects in liquid phase hydrogenations catalysed by supported metal nanoparticles. AB - PHIP signals have been observed, for the first time, in the heterogeneously catalysed liquid-phase hydrogenation of alkenes and alkynes using a range of supported metal nanoparticles on porous materials. PMID- 19562166 TI - Intramolecular apical metal-H-Csp3 interaction in molybdenum and silver complexes. AB - The reaction of HTIMP3 (HTIMP3=tris[1-diphenylphosphino)-3-methyl-1H-indol-2 yl]methane) with AgBF4 and Mo(CO)3(NCCH3)3 leads to Ag(HTIMP3)BF4 and Mo(CO)3(HTIMP3), respectively. The metal centre is coordinated to the three phosphorus atoms of the HTIMP3 ligand, which adopts a facial coordination mode, placing a H-Csp3 hydrogen atom at the apical position close to the metal centre. The solid-state structure of Mo(CO)3(HTIMP3) has been determined by X-ray crystallography, and the data have been used as input parameters for obtaining the optimised geometry of the complex using the B3PW91 functional. The silver structure has been modelled from the X-ray parameters of the molybdenum structure. In addition, theoretical calculations on the H-Csp3 downfield shift upon metal coordination has also been performed. They reproduce the experimental H-Csp3 chemical shifts well and supports that proton deshielding is mainly due to the presence of the metal, since the hydrogen is already located in the cone created by the aromatic-phosphino arms in the free ligand. PMID- 19562167 TI - Structural regularity and diversity in hybrids of aromatic thioethers and BiBr3: from discrete complexes to layers and 3D nets. AB - This paper aims to explore the regular structural patterns and the associated electronic properties of a group of hybrid networks based on BiBr3 and aromatic thioethers, with emphasis on structurally correlating the organic molecules with the BiBr3 aggregates as well as the overall hybrid networks. It was found that extended BiBr3 chains tend to be associated with the slender 4 methylthiophenylalkynyl units, whereas biaryl-based molecules with 4-MeSPh groups directly linked to aromatic cores form hybrids with discrete BiBr3 clusters of variable nuclearities and connectivities. By using multidentate ligands with open geometries, this exploratory study also achieves hybrid BiBr3-aromatic thioether networks with open framework and higher dimensional (e.g., 3D) features, which are potentially amenable to further study on guest exchange experiments. Diffuse reflectance measurements on the powder samples of the hybrids and the organic molecules reveal significant electronic interactions between the inorganic and organic components, with the absorption edges of the hybrids uniformly shifted to lower energies relative to the organic samples. In systems with similar network connectivity and local bonding features, the shifts appear to be proportional to the absorption energy of the organic molecules, suggesting that, in a first order approximation, the absorption edge in the hybrids involves substantially the electronic transitions from the HOMOs of the organic molecules to the LUMOs (or conduction bands) of the inorganic components. PMID- 19562168 TI - Molybdenum-oxide based unique polyprotic nanoacids showing different deprotonations and related assembly processes in solution. AB - We report the self-assembly processes in solution of three Keplerate-type molybdenum-oxide based clusters {Mo72V30}, {Mo72Cr30} and {Mo72Fe30} (all with diameters of approximately 2.5 nm). These clusters behave as unique weak polyprotic acids owing to the external water ligands attached to the non-Mo metal centers. Whereas the Cr and Fe clusters have 30 water ligands attached at the 30 M3+ centers pointing outside, {Mo72V30} has 20 water ligands coordinated to vanadium atoms, of which only 10 are pointing outside. The self-assembly processes of the Keplerates leading to supramolecular blackberry-type structures are influenced by the effective charge densities on the cluster surfaces, which can be tuned by the pH values and solvent properties. As expected, {Mo72Cr30} and {Mo72Fe30} behave similarly in aqueous solution due to their analogous structures and in both cases the self-assembly follows the partial deprotonation of the external water ligands attached to the non-Mo metal centers. However, the M-OH2 functionalities differ not only in acidity but also lability, i.e. in different residence times of the H2O ligands. In contrast to {Mo72Cr30} and {Mo72Fe30}, the {Mo72V30} clusters carry a rather large number of negative charges so that their solution properties are different. They exist as discrete macroions in dilute aqueous solution, and form only in mixed water/organic solvent (like acetone) blackberry-type structures whose size increases with acetone content. The comparison of the properties of the clusters allows more general information about the interesting self-assembly phenomenon to be unveiled. PMID- 19562169 TI - Chemoselective and biomimetic hydroxylation of hydrocarbons by non-heme micro-oxo bridged diiron(III) catalysts using m-CPBA as oxidant. AB - Three novel non-heme micro-oxo-bridged diiron(III) complexes [Fe2(micro-O)(L1)2] 2, where H2(L1) is N,N'-o-phenylenebis(salicylideneimine), [Fe2(micro O)(L2)2].2H2O 4, where H2(L2) is N,N'-o-phenylenebis(3,5-di-tert butylsalicylideneimine), and [Fe2(micro-O)(L3)2] 6, where H2(L3)=1,4-bis(2 hydroxybenzyl)-1,4-diazepane, have been isolated and studied as catalysts for the selective oxidative transformation of alkanes into alcohols using m choloroperbenzoic acid (m-CPBA) as co-oxidant. The mononuclear iron(III) complexes [Fe(L1)Cl] 1 and [Fe(L4)Cl] 7, where H2(L4)=1,4-bis(2-hydroxy-3,5-di tert-butylbenzyl)-1,4-diazepane, have been also isolated and those corresponding to the dimeric complexes 4 and 6 have been generated in CH3CN solution and characterized as [Fe(L2)Cl] 3 and [Fe(L3)Cl] 5 by using ESI-MS, absorption and EPR spectral and electrochemical methods. The molecular structures of 4 and 6 have been successfully determined by single crystal X-ray diffraction. Both 4 and 6 possess the Fe-O-Fe structural motif with each iron atom possessing a distorted square pyramidal coordination geometry. The steric constraint at the iron(III) center in 6 is higher than that in 4 as understood from the values of the trigonality structural index (tau: 4, 0.226, 0.273; 6, 0.449) due to the higher steric congestion built by the diazapane back bone. The micro-oxo-to-Fe(III) LMCT band for 4 is observed around 622 nm (epsilon, 1830 M(-1) cm(-1)) in methanol but is not observed in CH3CN solution and it is blue-shifted to around 485 nm (epsilon, 5760 M(-1) cm(-1)) in 6, possibly due to the higher Fe-O-Fe bond angle in the latter (4, 177.4; 6, 180 degrees). The Fe(III)/Fe(II) redox potentials of the dinuclear complexes (E1/2: 2, -0.606; 4, -0.329; 6, -0.889 V) are more negative than those for their corresponding mononuclear complexes (E1/2: 1, 0.300 V; 3, -0.269; 5, -0.289 V) due to O2- coordination. Interestingly, upon addition of peroxides (H2O2, t-BuOOH) and the peracid m-CPBA, the intensity of the phenolate-to-Fe(III) LMCT band for 2 and 6 decreases but does not exhibit any appreciable change for 4. In the presence of m-CPBA cyclohexane is selectively (A/K, 12.2) oxidized by the dimeric complex to cyclohexanol (A, CyOH) and a small amount of the further oxidized product cyclohexanone (K, CyO). However, interestingly, the corresponding monomeric complex affords enhanced yields of both CyOH and CyO but with a lower selectivity (A/K=1.7) and also 1 chlorocyclohexane via oxidative ligand transfer (OLT). The oxidation of adamantane by 4 affords exclusively 1-adamantanol (50.5%) and 2-adamantanol (9.5%) with enhanced yields over 12 h. In contrast, 3 provides 1-adamantanol (32.4%) and 2-adamantanol (14.8%) and adamantanone (14.6%) in addition to 1 chloroadamantane (14.1%) as the OLT product. The secondary C-H bond of ethylbenzene is randomly activated by both 3 and 4 to give 1-phenylethanol and acetophenone. Also, oxidation of cumene with tertiary C-H bonds to give 2-phenyl 2-propanol and the further oxidized product acetophenone is illustrated by invoking the iron-phenoxyl radical species as invoked for metalloporphyrin catalyzed systems. The strong chemoselectivity in C-H bond activation of alkanes by 4has been illustrated by invoking the involvement of a high-valent iron-oxo intermediate generated by using m-CPBA rather than the conventional oxidants H2O2 and t-BuOOH. In contrast to 4, the complexes 2 and 6 fail to effect the oxidation of hydrocarbons in the presence of H2O2, t-BuOOH and m-CPBA as the co-oxidant. PMID- 19562170 TI - Manganese complexes of phosphino-micro-phosphido ligands. AB - The reaction of the manganese precursors [MnOTf(CO)3{1,2-(PH2)2C6H4}], 5, or [MnOTf(CO)3{1,2-(PH2)2C2H4}], 6, with either the silver complex of 1,3 diallylbenzimidazol-2-ylidiene or the free carbene, resulted in the formation of the new, dimeric phosphino-micro-phosphido manganese species 7 and 8. Complex 7 crystallises in two isomeric forms (cis and trans), depending upon the choice of solvent. The structures of both isomers have been determined by single-crystal X ray techniques. PMID- 19562171 TI - Rational design of manganese-micro-azido 3D compounds by using diazines as co ligands: synthesis, structure and magnetic behaviour. AB - The syntheses, structural characterization, and magnetic behaviour of three new 3D manganese(II) complexes with the empirical formulae [Mn(N3)2(5-Brpym)]n (1), [Mn(N3)2(Mepyz)]n (2) and [Mn(N3)2(2-acpy)]n (3), (5-Brpym=5-bromopyrimidine, Mepyz=methylpyrazine, 2-acpy=2-acetylpyridine), are reported. In 1, each manganese atom is linked to the four nearest neighbours by only end-to-end azido bridges, forming square layers. These layers are further connected to 3D networks by the N,N'-bridging ligand. In 2, Mn2(micro(1,1)-N3)2 subunits are connected to the four nearest neighbours by end-to-end azido bridges to form a 3D Mn-azido sublattice. The Mn2(micro(1,1)-N3)2 subunits are further linked by pairs of N,N' bridging Mepyz ligands to form 1D ribbons extended along the c-axis of the unit cell. The Mepyz pairs show pi-pi-interactions. In 3, the only end-to-end azido bridges form a diamondoid-like 3D Mn-azido-sublattice. The magnetic properties of 1-3 are reported. At high temperatures, the plots of chiM or chiMTvs.T for can be fitted as a homogeneous 2D system with J=-5.4 cm(-1), g=2.05. Also, at high temperatures the plots of chiM or chiMTvs.T for compound can be fitted with the simple cubic expansion series with J=-1.6 cm(-1), g=2.04. Compound 1 shows spontaneous magnetization below Tc=45 K, which corresponds to the presence of spin-canted antiferromagnetism, whereas 2 and 3 do not show spontaneous magnetization up to 2 K. PMID- 19562172 TI - 182tungsten Mossbauer spectroscopy of heteropolytungstates. AB - The tungsten-182 Mossbauer spectra of a series of Keggin structure heteropolytungstates, [EW12O40]n- are reported. There is a very considerable variation in quadrupole coupling at the tungsten nucleus indicating considerable asymmetry in the electron distribution for the more electronegative elements E. The quadrupole coupling correlates well with the structural data, in particular with the distance between the tungsten and the oxygen atom of the EO4 group. These compounds may be regarded as rigid W12O36 cages interacting more or less strongly with an EO4n- host. The spectra of salts of metatungstate [H2W12O40]6- and [W6O19]2- are also given. PMID- 19562173 TI - Synthesis, characterization, in vivo antitumor properties of the cluster rhenium compound with GABA ligands and its synergism with cisplatin. AB - A new dirhenium(III) complex cis-[Re2(GABA)2Cl5(H2O)]Cl.2H2O with zwitterionic gamma-aminobutyrate ligands was prepared and characterized by spectral methods and crystallography. The structure of the compound is comprised of dinuclear complex cations (Re-Re 2.2437(3) A) involving cis-oriented double carboxylate bridges, four equatorial chloride ions and two weakly bonded aqua and chloride ligands in the axial positions at two rhenium centers (Re-O 2.363(3), Re-Cl 2.6735(12) A). Antitumor properties of the complex were studied in the model of tumor growth with the use of Wistar rats inoculated by tumor carcinoma Guerink cells. The introduction of the compound in dosage according to the scheme of antioxidant therapy, inhibited the tumor growth by ca. 60% and led to stabilization of red blood cells in the tumor-bearing organisms. The combined introduction of the compound and cisplatin had a significant impact on the tumor growth and the disappearance of the tumors in most of the animals. PMID- 19562174 TI - Synthesis, structures and characterisations of truly homoleptic acetonitrile Ln3+ complexes in solid state and in solution. AB - Total halide abstraction from LnCl3 by Ag[Al(OC(CF3)3)4]/CH3CN has been confirmed for a series of lanthanide metal ions by the structural characterization of [Ln(CH3CN)n][Al(OC(CF3)3)4]3 (n=9, Ln3+=Nd, Eu, Gd, Dy; n=8, Ln3+=Tm) complexes. Evidence for the very low coordinating ability of the [Al(OC(CF3)3)4]- anion towards Ln3+ ions is provided in the solid state (X-ray, IR and Raman spectroscopy) and in anhydrous acetonitrile solution (conductivity, EPR and NMR measurements). In the solid state homoleptic nine-coordinated acetonitrile species are characteristic for lanthanides for the beginning (Nd) and the middle (Eu, Gd, Dy) of the Ln series, with a mono-capped square antiprismatic arrangement of the N donor atoms around the metal centres; while for those from the end of the series (Tm) eight-coordinated species are representative with a square antiprismatic arrangement. In anhydrous acetonitrile solution, conductivity measurements revealed 3:1 electrolyte types for all compounds. EPR and 19F NMR line broadening measurements attest lanthanide complexes free of any coordinating [Al(OC(CF3)3)4]- anion. PMID- 19562175 TI - Rhenium and 99m-technetium complexes of monosaccharide based tripodal triamines as potential radio imaging agents. AB - A synthetic pathway to new sugar containing tripodal triamines of the TAME type (1,1,1-tris(aminomethyl)ethane) is presented. The tripodal bromo substituted precursors Ac3Xyl-O-CH2C(CH2Br)3, Ac4Glc-O-CH2C(CH2Br)3 and Ac4Gal-O-CH2C(CH2Br)3 (2a-c) were obtained by glycosidation reaction of the fully acetylated glycopyranosides with pentaerythritol tribromide. Nucleophilic substitution to the corresponding azides with sodium azide and deprotection of the sugars, followed by hydrogenation reaction in the presence of PtO2 leads to the triamines Xyl-O-CH2C(CH2NH2)3, Glc-O-CH2C(CH2NH2)3 and Gal-O-CH2C(CH2NH2)3 (5a-c). The triamines form complexes of the type [Re(CO)3L]Cl (6a-c). The precursors as well as the final ligands and complexes were characterized by elemental analysis, IR and NMR spectroscopy as well as mass spectrometry. The synthesis of the analogous radiolabelled 99mTc complex with galactosyl appendage 7c (LGal-O-CH2C(CH2NH2)3) was achieved for 5c and its stability over a period of 24 h could be verified by HPLC analysis, confirming the significant stability of 7c against histidine exchange. PMID- 19562176 TI - Molecules based on M(v) (M=Mo, W) and Ni(II) ions: a new class of trigonal bipyramidal cluster and confirmation of SMM behavior for the pentadecanuclear molecule {NiII[NiII(tmphen)(MeOH)]6[Ni(H2O)3]2[micro-CN]30[WV(CN)3]6}. AB - The preparation, single crystal X-ray crystallography, and magnetic properties are reported for four new clusters based on [M'V(CN)8]3- octacyanometallates (M'=Mo, W). Reactions of [M'V(CN)8]3- with mononuclear NiII ions in the presence of the tmphen blocking ligand (tmphen=3,4,7,8-tetramethyl-1,10-phenanthroline) in a 2:3:6 ratio, respectively, lead to the formation of the trigonal bipyramidal clusters [NiII(tmphen)2]3[M'V(CN)8]2. Analogous reactions with the same starting materials performed in a 2:3:2 ratio, respectively, produce pentadecanuclear clusters of the type {NiII[NiII(tmphen)(MeOH)]6[Ni(H2O)3]2[micro CN]30[WV(CN)3]6}. The W2Ni3 (1) and Mo2Ni3(2) pentanuclear clusters and the W6Ni9 (3) and Mo6Ni9 (4) pentadecanuclear molecules are isostructural to each other and crystallize in the space groups P2(1)/c and R3 respectively. Magnetic measurements indicate that the ground states for the trigonal bipyamidal clusters are S=4 as a consequence of ferromagnetic coupling with JW-Ni=9.5 cm(-1), JMo Ni=10 cm(-1). The pentadecanuclear clusters exhibit ferromagnetic coupling as well, which leads to S=12 ground states (JW-Ni=12 cm(-1), JMo-Ni=12.2 cm(-1)). Reduced magnetization studies on the W-Ni analogues support the conclusion that they exhibit a negative axial anisotropy term; the fits give D values of -0.24 cm(-1) for the W2Ni3 cluster and D=-0.04 cm(-1)for the W6Ni9 cluster. AC susceptibility measurements indicate the beginning of an out-of-phase signal for the W2Ni3 and the W6Ni9 compounds, but detailed low temperature studies on small crystals by the microSQUID technique indicate that only the pentadecanuclear cluster exhibits hysteresis in accord with SMM behavior. Neither Mo cluster reveals any evidence for slow paramagnetic relaxation at low temperatures. PMID- 19562177 TI - Geometrical isomers of [TEAH][Co(LSe)2].xH2O: synthesis, structural, spectroscopic and computational studies. AB - Trans-1, [HNEt3][CoIII(LSe)2].H2O and cis-1, [HNEt3][CoIII(LSe)2].3H2O have been synthesized and characterized by single-crystal X-ray studies. The counter ion Et3NH+ plays a crucial role in the crystal packing leading to the formation of two distinctly different supramolecular assemblies in the two complexes. In trans 1, Co-bisphenolate units and triethylamine molecules are arranged in a linear fashion leading to a supramolecular columnar assembly along the crystallographic a-axis. In this assembly, triethylammonium ions are sandwiched between successive Co-bisphenolate units and act as gluing agents joining Co-bisphenolate units on either side through C-Hpi interactions. In sharp contrast to trans-1, Co bisphenolate units and triethylammonium ions in cis-1 are arranged in a helical supramolecular assembly through similar C-Hpi interactions along the crystallographic b-axis. The SeSe van der Waals interactions may be responsible for the predominant occurrence of the cis- isomer. The cyclic voltammetric studies showed quasi-reversible waves for the cobalt(III)-->cobalt(II) reductions with E1/2=0.635 and 0.628 V vs. Ag/AgCl for cis-1 (at approximately 5 degrees C) and trans-1 (at approximately 25 degrees C), respectively. DFT calculations show that the trans-form is the thermodynamic product with higher stability than the cis- one, which is consistent with the variable temperature 1H NMR studies. PMID- 19562178 TI - A dipalladium complex with a single hydroxo bridge and its methylpalladium precursor. AB - The reaction of [PdClMe(Me2NCH2CH2NMe2)] with bis(2-pyridylmethyl)amine, bpma, gave the methylpalladium complex [PdMe(bpma)]Cl, which could be converted to the triflate or tetrafluoroborate salt by reaction with AgOTf or AgBF4. Further reaction of [PdMe(bpma)]OTf with B(C6F5)3 or HOTf gave the complex [Pd2(micro OH)(bpma)2](OTf)3, which contains a bridging hydroxo ligand and no other bridging ligand, and whose structure contains an unexpectedly large Pd-O-Pd angle of 141.2(3) degrees. This complex acts as a model for complexes proposed to be formed during hydrolysis in aqueous solution of biologically active compounds such as cisplatin. PMID- 19562179 TI - Cation ordering in Li containing garnets: synthesis and structural characterisation of the tetragonal system, Li7La3Sn2O12. AB - In this paper we report synthesis, conductivity and structural data for the novel tetragonal garnet-related system, Li7La3Sn2O12. Neutron diffraction data shows that the tetragonal distortion is related to ordering of Li in three sites within the structure to ensure no short Li-Li interactions. Consistent with the ordered nature of the Li ions, the conductivity is low, with a high activation energy. The results are relevant to related highly conducting cubic garnets, Li5+xLn3 xAxM2O12 (Ln=rare earth, A=alkaline earth; M=Nb, Ta, Sb), showing how a high Li content can be accommodated by Li ordering within the garnet structure, supporting previous suggestions by Cussen for the cubic garnets, who proposed the presence of local ordering/clustering of Li in tetrahedral and "octahedral" sites to limit unfavourable short Li-Li interactions. PMID- 19562180 TI - The intramolecular sp2 and sp3 C-H bond activation of (p-cymene)ruthenium(II) N heterocyclic carbene complexes. AB - The intramolecular sp2 and sp3 C-H activated products, as well as the monometalated products, based on the "(p-cymene)Ru(NHC)" framework were synthesised by treatment of a series of NHCs (1-R-3-methylimidazol-2-ylidene [R=Ph (1), Bn (2), t-Bu (3), i-Pr (4), Mes (5), Cy (6)] and 1,3 bis(isopropyl)imidazol-2-ylidene (7)) with [(p-cymene)RuCl2]2 under mild conditions. A new NHC precursor (1-tert-butyl-3-phenyl-4,5-dihydro-imidazol-2 ylidene) was also designed to compare the reactivity of sp2 C-H and sp3 C-H bonds upon cyclometalation, and only the sp3 C-H activated product (8) was observed. The factors that possibly determine the selectivity of intramolecular sp2 or sp3 C-H activation are elucidated by a series of experiments. In the cases where activation of both sp2 C-H and sp3 C-H is possible, steric factors overrode the others to dominate the regioselectivity of activation. All complexes were characterised by 1H NMR, 13C NMR and HRMS spectra. The molecular structures of 1, 3, 5, 6, 7, and 8 were confirmed by X-ray diffraction. PMID- 19562181 TI - Synthesis, cytotoxicity and cucurbituril binding of triamine linked dinuclear platinum complexes. AB - The platinum complexes trans-[{PtCl(NH3)2}2(micro-NH2(CH2)3NH2(CH2)3NH2)]3+ (CT033) and the corresponding N4-dimethyl linked analogue trans [{PtCl(NH3)2}2(micro-NH2(CH2)3N(Me)2(CH2)3NH2)]3+ (CT233) have been synthesised, and their cytotoxicity, ability to bind cucurbit[7,8]uril (Q[7,8]) and reaction with cysteine studied. Both platinum complexes show good activity in the L1210 cell line and maintain their activity in the corresponding cisplatin L1210/DDP cell line. However, the N4-dimethyl analogue CT233 is approximately 50-times less active than the CT033 complex. This suggests that the insertion of a positive charge into the linking ligand may not, per se, be responsible for the higher cytotoxicity generally observed for dinuclear platinum complexes linked by polyamines. The upfield shifts of the resonances from the methylene protons in the linking triamine ligand observed in the 1H NMR spectra of either CT033 and CT233 upon addition of either Q[7] or Q[8] indicate that the cucurbituril is positioned over the linking ligand. However, the results show that the protonated secondary amine in CT033 acts as a barrier to encapsulation, with the Q[7,8] being positioned over only one propyl-arm at a time. Alternatively, the entire triamine linking ligand of CT233 is fully encapsulated within the Q[7,8] cavity. Encapsulation by Q[7,8] was found to reduce the rate of reaction of CT033 and CT233 with the thiol containing amino acid cysteine, with a greater rate reduction observed for CT233. These results are consistent with the NMR results of the Q[7,8] binding studies of the two platinum complexes. For CT033 encapsulated in Q[7,8], one of the two platinum centres is completely exposed to the solvent, whereas, for CT233 both platinum centres are simultaneously positioned within the portals of the cucurbit[n]uril, thereby, affording greater protection. PMID- 19562182 TI - Bimetallic Schiff base complexes: models for conjugated shape-persistent metallopolymers. AB - New Schiff base ligands with two metal binding sites have been prepared. Copper and zinc complexes of the ligands, which serve as models for rigid, conjugated metallopolymers, were synthesized and characterized. The copper complexes display only weak intramolecular antiferromagnetic interactions, suggesting that the polymer structure is not useful for developing magnetic materials. Preliminary investigations of the novel polymers, including the preparation of a conjugated zinc-containing polymer, are reported. PMID- 19562183 TI - Synthesis, X-ray structures and reactivity of calix[5]arene bismuth(iii) and antimony(III) complexes. AB - A series of calix[5]arene bismuth(III) and antimony(III) mono- and bimetallic complexes were synthesized and fully characterized by NMR, X-ray, IR, mp, UV-Vis and elemental analysis. Reaction of p-tert-butylcalix[5]arene (tBuC5(H)5) trianionic salts M'3.tBuC5(H)2 (M'=Li, Na, K) with MCl3 (M=Bi, Sb) yielded monometallic complexes [Bi{tBuC5(H)2}] 1 and [Sb{tBuC5(H)2}] 2, respectively. 1H NMR spectra of both complexes showed two remaining OH groups available for further reactivity. Alternatively, complexes 1 and 2 can be obtained by reacting tBuC5(H)5 in a 1:1 ratio with M(OtBu)3, but the yields are lower. When the tBuC5(H)5 lower rim monobenzyl ether [tBuC5(Bn)(H)4] is treated in a 1:1 ratio with Bi(OtBu)3, the monometallic complex [Bi{tBuC5(Bn)(H)}]2 3 is prepared. If, however, [tBuC5(Bn)(H)4] reacts with Sb(NMe2)3 or Sb(OtBu)3 in a 1:2 ratio the production of the bimetallic complex [Sb2O{tBuC5(Bn)}] 4 is observed. p Benzylcalix[5]arene (BnC5(H)5) reacts with excess Bi(OtBu)3 to produce the bimetallic complex [Bi2O{BnC5(H)}]2 5. 1H NMR spectra of 5 display patterns characteristic for a cone conformer in solution. Treatment of calix[5]arene [HC5(H)5] with one equivalent of Bi[N(SiMe3)2]3 or with 0.75 equivalents of Sb(NMe2)3 yields bimetallic complexes [Bi2O{HC5(H)}] 6 and [Sb2O{HC5(H)}] 7, respectively. The reactivity of monometallic complexes 1 and 2 was tested in order to investigate the availability of their remaining OH groups. Treatment of 1 with Bi(OtBu)3 at ambient temperature yields bimetallic complex [Bi2O{tBuC5(H)}]2 8 while the reaction of complex 2 with Sb(OtBu)3 in a 1:1 ratio produces complex [Sb2O{tBuC5(H)}] 9. The crystal structures of the monometallic bismuth complexes 1 and 3 display dimeric units with the calixarene ligands in distorted cone and "paco-in" conformations, respectively. Complexes 4, 7 and 9 are all monomeric units, displaying [(RO)2Sb]2(micro-O) cores and the calixarene ligands in 1,2-alternate conformation. The dimeric units of bimetallic complexes 5 and 8 contain Bi4O2(OR)8 core structures that force the calixarene ligands to adopt a flattened cone conformation. PMID- 19562184 TI - Synthesis of bismuth and antimony complexes of the "larger" calix[n]arenes (n=6 8); from mononuclear to tetranuclear complexes. AB - A series of calix[n]arene (n=6-8) bismuth and antimony complexes were synthesized and fully characterized by NMR, X-ray, IR, UV-Vis and elemental analysis. The monobismuth calix[6]arene complex [Bi{tBuC6(H)3}]2 1 was prepared by the reaction of para-tert-butylcalix[6]arene (tBuC6(H)6) with one equivalent of Bi[N(SiMe3)2]3. Complex 1 featured a Bi2(micro-O)2 central core similar to other bismuth calixarene complexes prepared by our group. Reaction of calix[6]arene (HC6(H)6) with two equivalents of Bi(OtBu)3 yielded different outcomes depending on the reaction solvent. If THF was used, complex [Bi{HC6(H)3}] 2 was obtained in 72% yield; however, when toluene was used, complexes 2 and [Bi2{HC6}] 3 were isolated in 23 and 57% yields, respectively. Mononuclear complexes 1 and 2 displayed dimeric structures in the solid state with cone-like conformations for the calixarene ligands. The 1H NMR spectrum of complex 2 displays patterns for an asymmetric structure with two signals in a 2:1 ratio for the unreacted OH groups. Treatment of calix[6]arenes RC6(H)6 (R=H, tBu) with two equivalents of SbR3 (R=OtBu, NMe2) produced dinuclear complexes [Sb2{HC6}] 4, and [Sb2{tBuC6}] 5, respectively. The 1H NMR spectra for the dinuclear complexes 3, 4, and 5 showed the characteristic calixarene pattern for a 1,2,3-alternate conformer. In the process of recrystallization of complex 4 an unexpected trimetallic complex with composition [Sb3O2{HC6(H)}] 4a was obtained in low yield. Treatment of para-tert butylcalix[7]arene (tBuC7(H)7) with two equivalents of Bi(OtBu)3 produced the bimetallic complex [Bi2O{tBuC7(H)3}]2 6. Complex 6 contains an overall Bi4O2(OAr)8 core system with a structural resemblance to other bimetallic bismuth calixarene complexes reported by our group. The larger para-benzylcalix[8]arene (BnC8(H)8) and calix[8]arene (HC8(H)8) reacted with excess Bi(OtBu)3 to produce the tetranuclear complexes [Bi4O2{HC8}] 7 and [Bi4O2{BnC8}] 8, respectively. The solid state structure of complex 8 featured a dimeric unit with the calixarene ligands in pinched-cone conformation and displaying an overall Bi8O4(OAr)16.H2O core. No metal pi-arene interactions were observed. PMID- 19562188 TI - ps-TRIR covers all the bases--recent advances in the use of transient IR for the detection of short-lived species in nucleic acids. AB - Recent developments of the picosecond transient absorption infrared technique and its ability to elucidate the nature and kinetic behaviour of transient species formed upon pulsed laser excitation of nucleic acids are described. PMID- 19562189 TI - Micro-electromechanical sensors in the analytical field. AB - Micro- and nano-electromechanical systems (MEMS and NEMS) for use as sensors represent one of the most exciting new fields in analytical chemistry today. These systems are advantageous over currently available non-miniaturized sensors, such as quartz crystal microbalances, thickness shear mode resonators, and flexural plate wave oscillators, because of their high sensitivity, low cost and easy integration into automated systems. In this article, we present and discuss the evolution in the use of MEMS and NEMS, which are basically cantilever-type sensors, as good analytical tools for a wide variety of applications. We discuss the analytical features and the practical potential of micro(nano)-cantilever sensors, which combine the synergetic advantages of selectivity, provided by their functionalization, and the high sensitivity, which is attributed largely to the extremely small size of the sensing element. An insight is given into the different types of functionalization and detection strategies and a critical discussion is presented on the existing state of the art concerning the applications reported for mechanical microsensors. New developments and the possibilities for routine work in the near future are also covered. PMID- 19562190 TI - An operationally simple colorimetric assay of hyaluronidase activity using cationic gold nanoparticles. AB - An operationally simple colorimetric way for measuring hyaluronidase activity was developed using cysteamine-bound gold nanoparticles. The addition of gold nanoparticles into hyaluronidase-containing solutions resulted in color changes, which could easily be observed with the naked eye or a UV/Vis spectrophotometer. PMID- 19562191 TI - An immunoassay using biotinylated single-walled carbon nanotubes as Raman biomarkers. AB - A new immunoassay with biotinylated single-walled carbon nanotubes as persistent, non-photobleaching Raman biomarkers demonstrated excellent sensitivity and specificity. PMID- 19562192 TI - Chemiluminescence from reactions with bis-cyclometalated iridium complexes in acidic aqueous solution. AB - Chemical reactions between certain bis-cyclometalated iridium complexes, cerium(IV) and organic reducing agents in aqueous solution produce an emission of light which in some cases is more intense than that from analogous reactions with conventional ruthenium-based reagents, thus providing a new avenue for chemically initiated luminescence detection. PMID- 19562193 TI - Separation of oligonucleotides with single-base mutation by capillary electrophoresis using specific interaction of metal ion with nucleotide. AB - A unique tactic for the separation of single-base sequential isomers of oligomeric single-stranded DNA by a CE separation system employing the specific interaction of metal ion with nucleotide was demonstrated, enabling the separation of the mixture of a 12-mer oligonucleotide and its single-base mutants, as well as their positional isomers. PMID- 19562194 TI - Scanning electrochemical microscopy of genomic DNA microarrays--study of adsorption and subsequent interactions. AB - The adsorption of genomic DNA and subsequent interactions between adsorbed and solvated DNA have been studied using scanning electrochemical microscopy (SECM). Microarrays of polyethylenimine (PEI) films could be deposited on screen-printed carbon substrates using the SECM. Single stranded herring DNA was electrostatically adsorbed at the surface of the polyethylenimine. The further adsorption of complementary single stranded DNA on the surface was observed to give rise to substantial decreases in interfacial impedance at the surface as measured by increases of tip current of the order of 1-2 nA (6%). Conversely adsorption of DNA from alternate species, i.e. salmon ssDNA on herring ssDNA, yielded much smaller changes in tip current of 0.2 nA. The significance of this work is that the approach opens up the possibility for direct label-free electrochemical interrogation of DNA microarrays as an alternative to other existing optical techniques. PMID- 19562195 TI - Interaction of metal ions and DNA films on gold surfaces: an electrochemical impedance study. AB - Electrochemical impedance spectroscopy (EIS) has been used to investigate the effects of a number of metal ions with DNA films on gold surfaces exploiting [Fe(CN)6](3-/4-) as a solution-based redox probe. Alkaline earth metal ions Mg2+, Ca2+, trivalent Al3+, La3+ and divalent transition metal ions Ni2+, Cu2+, Cd2+ and Hg2+ have been selected in this study and the results are compared with previous studies on the effects of Zn2+ on the EIS of DNA films. All experimental results were evaluated with the help of equivalent circuits which allowed the extraction of resistive and capacitive components. For all metal ions studied here, addition of the metal ions causes a decrease in the charge transfer resistance. The difference of charge transfer resistance (DeltaR(ct)) of ds-DNA films in the presence and absence of the various metal ions is different and particular to any given metal ion. In addition, we studied the EIS of ds-DNA films containing a single A-C mismatch in the presence and absence of Ca2+, Zn2+, Cd2+ and Hg2+. DeltaR(ct) values for ds-DNA films with a single A-C mismatch is smaller than those of fully matched ds-DNA films. PMID- 19562196 TI - Using scanning contactless conductivity to optimise photografting procedures and capacity in the production of polymer ion-exchange monoliths. AB - Capacitively coupled contactless conductivity detection (C4D) is utilised as a simple, rapid and non-invasive technique for the quantitative evaluation of the ion-exchange capacity of charged polymer monoliths in capillary format. A charged monomer, 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) was photografted onto a 100 microm i.d. butyl methacrylate-co-ethylenedimethacrylate monolith in a number of discrete 10 mm zones. By varying the energy dose (J/cm2) during grafting of each zone, the grafting density and thus ion-exchange capacity could be precisely controlled. Ion-exchange capacity could be correlated with energy dose by measuring the conductive response of each grafted region using scanning C4D techniques. Repeatability of the scanning C4D method was excellent with % RSD values of 0.7% and 2.4% obtained for three replicate scans of the ungrafted and grafted regions of a single monolith, respectively. Repeatability of the photografting process on separate monoliths was also examined by comparison of C4D profiles. The spatial accuracy of photografting was probed using scanning C4D which could measure the conductive response of the monolith at measurement intervals as low as 1 mm along its entire length. Scanning C4D was also used for the real time visualisation of the equilibration of grafted zones to permit the optimisation of monolith washing procedures. Finally, scanning C4D was applied to the measurement of the ion-exchange capacity of butyl methacrylate-co-AMPS-co ethylenedimethacrylate copolymers with a direct correlation between monolith conductive response and concentration of charged monomer in the polymerisation mixture. The longitudinal homogeneity of charge along the monolith was 0.3% RSD, demonstrating that the charged functional monomer was evenly dispersed throughout the bulk of the monolith. Ion-exchange capacity was cross validated chromatographically using breakthrough studies and found to closely correlate to within 1% of the measurements made by scanning C4D. PMID- 19562197 TI - Mass spectrometry tools and metabolite-specific databases for molecular identification in metabolomics. AB - The chemical identification of mass spectrometric signals in metabolomic applications is important to provide conversion of analytical data to biological knowledge about metabolic pathways. The complexity of electrospray mass spectrometric data acquired from a range of samples (serum, urine, yeast intracellular extracts, yeast metabolic footprints, placental tissue metabolic footprints) has been investigated and has defined the frequency of different ion types routinely detected. Although some ion types were expected (protonated and deprotonated peaks, isotope peaks, multiply charged peaks) others were not expected (sodium formate adduct ions). In parallel, the Manchester Metabolomics Database (MMD) has been constructed with data from genome scale metabolic reconstructions, HMDB, KEGG, Lipid Maps, BioCyc and DrugBank to provide knowledge on 42,687 endogenous and exogenous metabolite species. The combination of accurate mass data for a large collection of metabolites, theoretical isotope abundance data and knowledge of the different ion types detected provided a greater number of electrospray mass spectrometric signals which were putatively identified and with greater confidence in the samples studied. To provide definitive identification metabolite-specific mass spectral libraries for UPLC-MS and GC-MS have been constructed for 1,065 commercially available authentic standards. The MMD data are available at http://dbkgroup.org/MMD/. PMID- 19562198 TI - UV-PEDD photometry dedicated for bioanalytical uses. AB - Detection of p-nitrophenol, a product of several enzymatic reactions, is useful and important in many fields of modern bioanalytics. Here, we have developed and applied a complete photometer dedicated for such measurements that is extremely economic and easily customized for specific experimental setups. The instrument consists only of paired emitter-detector diodes (PEDDs) coupled with an extremely simple and low-cost signal transduction system. The experimental data confirmed that this photometric system adapted for cuvette as well as flow measurements can be applied for fast and reproducible determination of alkaline phosphatase (ALP) activity. PMID- 19562199 TI - Circular arrays of polymer-based miniature rectilinear ion traps. AB - The design and operation of an annular array of parallel, miniature rectilinear ion traps (RITs) is discussed. Stereolithography apparatus (SLA), a previously validated method for ion trap fabrication, was applied here to construct an array of mass analyzers and their mounting hardware. Two versions of the array were tested, using either six or twelve stretched RITs (x0 = 1.66 mm, y0 = 1.33 mm, z = 16.66 mm) mounted in parallel about the circumference of a circle with the interior and exterior x-electrode planes oriented tangential to the inner and outer annulus rings, respectively. The arrangement of the ion traps is such that the ions are radially ejected just above the throat of a centrally located electron multiplier detector into which they are accelerated. The mass analyzer array was mounted in a custom vacuum manifold. The resolution, mass-to-charge ratio (m/z) range, and MS/MS capabilities were tested using electrospray ionization (ESI). The devices were tested in two configurations: (i) separate ion sources for each trap, and (ii) a single ion source for the entire array. PMID- 19562200 TI - Ultrasensitive Pb2+ detection based on fluorescence resonance energy transfer (FRET) between quantum dots and gold nanoparticles. AB - Positively charged CdTe-QDs capped with cysteamine (CA-CdTe-QDs) and negatively charged AuNPs capped with 11-mercaptoundecanoic acid (MUA-AuNPs) have been prepared. They are water-soluble and biocompatible. An assay for the determination of Pb2+ has been proposed based on the modulation in FRET efficiency between QDs and AuNPs in the presence of Pb2+, which inhibits the interaction of the QD-AuNP assembly. This method is easy to operate and with remarkably high sensitivity. Under the optimum conditions, the response is linearly proportional to the concentration of Pb2+ in the range 0.22-4.51 ppm, and the detection limit is found to be 30 ppb of Pb2+ due to the superior fluorescence properties of QDs. The mechanism of this strategy is also discussed. PMID- 19562201 TI - Adenosine detection by using gold nanoparticles and designed aptamer sequences. AB - Based on gold nanoparticles (AuNPs) and engineered DNA aptamers, we designed a novel bioassay strategy for the detection of adenosine as a small target molecule. In this design, an aptamer is engineered to consist of two pieces of random-coil like ssDNA which are respectively attached to AuNPs through their 5' thiol-modified end. They can reassemble into the intact aptamer tertiary structure and induce nanoparticle aggregation in the presence of the specific target. Results have demonstrated that gold nanoparticles can effectively differentiate these two different DNA structures via their characteristic surface plasmon resonance-based color change. With this method, adenosine can be selectively detected in the low micromolar range, which means that the strategy reported here can be applicable to the detection of several other small target molecules. PMID- 19562202 TI - Sensitive and selective detection of cysteine using gold nanoparticles as colorimetric probes. AB - We report herein the development of a highly sensitive and selective colorimetric detection method for cysteine using gold nanoparticles probes. This assay relies upon the distance-dependent optical properties of gold nanoparticles, the self assembly of cysteine on gold nanoparticles, and the interaction of a 2:1 cysteine/Cu2+ complex. In the presence of Cu2+, cysteine could rapidly induce the aggregation of gold nanoparticles, thereby resulting in red-to-blue (or purple) color change. The concentration of cysteine can be determined by monitoring with the naked eye or a UV-vis spectrometer. The present limit of detection for cysteine is 10 nM. This method exhibits excellent selectivity for cysteine over other alpha-amino acids, glutathione, thioglycolic acid and mercaptoethyl alcohol. PMID- 19562203 TI - Semi-quantitative analysis of the monomer composition in co-polymer microgels using solid state Raman and NMR spectroscopy. AB - A series of colloidal microgels have been prepared by surfactant-free emulsion polymerisation (SFEP) based on the N-isopropylacrylamide (NIPAM) monomer. 4 Vinylpyridine (4-VP) and butylacrylate (BuA) have been used as co-monomers. Co polymer poly(NIPAM/4-VP) and poly(NIPAM/BuA) have been prepared with various monomer ratios, ranging from pure poly(NIPAM) to pure poly(BuA)/poly(4-VP). Freeze-dried samples of the microgels have been analysed by solid state (ss) Raman and NMR (Nuclear Magnetic Resonance) spectroscopy to investigate the monomer composition in the co-polymer microgels. Spectral data have been analysed graphically and also statistically. Spectroscopic measurements have shown that co polymerization has occurred. The graphical and statistical analysis of the spectroscopic data for both co-polymer microgels, enables the semi-quantitative measurement of the percentage incorporation of co-monomers (4-VP/BuA) in the co polymer microgels. A good correlation exists between the Raman and NMR results, however, Raman spectroscopy is much less time consuming (Raman spectral acquisition time is less than 10 minutes) and the measurements are easy to make and very small quantities (less than 1 mg) of the sample are required. This compares with the experimental measurements of approximately 72 hours and 100-200 mg of sample that are required for the NMR experiments. PMID- 19562204 TI - Targeting and detecting cancer cells using spontaneously formed multifunctional dendrimer-stabilized gold nanoparticles. AB - We develop a facile approach to fabricating multifunctional dendrimer-stabilized gold nanoparticles (Au DSNPs) for cancer cell targeting and imaging. In this work, amine-terminated generation 5 (G5) poly(amidoamine) (PAMAM) dendrimers pre functionalized with folic acid (FA) and fluorescein isothiocyanate (FI) are complexed with Au(III) ions, followed by acetylation of the amine groups on the dendrimer surfaces. This one-step process leads to the spontaneous formation of 6 nm-sized Au nanoparticles stabilized by multifunctional dendrimers bearing both targeting and imaging functionalities. The multifunctional Au DSNPs are characterized by UV-Vis spectrometry, 1H NMR, and transmission electron microscopy (TEM). The formed Au DSNPs are water-soluble, stable, and biocompatible. Combined flow cytometry, confocal microscopy, silver staining, and inductively coupled plasma-mass spectrometry (ICP-MS) analyses show that the FA- and FI-functionalized Au DSNPs can specifically target to cancer cells expressing high-affinity FA receptors in vitro. This approach to functionalizing Au DSNPs may be extended to other targeting molecules, providing a unique nanoplatform for targeting and imaging of a variety of biological systems. PMID- 19562206 TI - Optofluidic ring resonator sensors for rapid DNT vapor detection. AB - We demonstrated rapid 2,4-dinitrotoluene (DNT) vapor detection at room temperature based on an optofluidic ring resonator (OFRR) sensor. With the unique on-column separation and detection features of OFRR vapor sensors, DNT can be identified from other interferences coexisting in the analyte sample mixture, which is especially useful in the detection of explosives from practical complicated vapor samples usually containing more volatile analytes. The DNT detection limit is approximately 200 pg, which corresponds to a solid phase microextraction (SPME) sampling time of only 1 second at room temperature from equilibrium headspace. A theoretical analysis was also performed to account for the experimental results. Our study shows that the OFRR vapor sensor is a promising platform for the development of a rapid, low-cost, and portable analytical device for explosive detection and monitoring. PMID- 19562205 TI - Dithizone nanofiber-coated membrane for filtration-enrichment and colorimetric detection of trace Hg(II) ion. AB - Dithizone nanofiber-coated membranes (dithizone membranes), which are useful for sensitive and selective determination of Hg(II), were fabricated. Simply by filtration of the aqueous dispersion of dithizone nanofiber through a cellulose ester membrane filter, a dithizone nanofiber layer of less than 500 nm thickness was coated firmly and uniformly over the membrane filter surface. The steel blue color of the membrane remained unchanged for more than three months when fabricated in the presence of ascorbic acid and stored with an oxygen absorber in an evacuated aluminium bag. Determination at the parts per billion level of Hg(II) was achieved by filtration-enrichment of a sample solution and simultaneous colorimetric analysis using a TLC scanner (500 nm). Consequently, Hg(II) ion was concentrated in the dithizone layer as reddish brown complexes by filtration of a sample solution at pH 2.7. More than 90% of 10 ppb Hg(II) was retained in the dithizone layer at the filtration rate of 1.3-9.3 ml min(-1). The presence of Na+ (10,000 ppm), K+ (5000 ppm), Ca(II) (5000 ppm), Cu(II) (6.4 ppm), Fe(II) (100 ppm), Zn(II) (100 ppm), Pb(II) (100 ppm) and Cd(II) (10 ppm) by using 2.5 x 10(-4) M of ethylenediamine tetraacetic acid (EDTA) as a masking reagent did not interfere with the detection of Hg(II) (10 ppb). Most of anions did not interfere with the determination of Hg(II). The present method was tested for the detection of simulated wastewater, river water and seawater spiked with 10 ppb of Hg(II). PMID- 19562207 TI - Electrochemical behavior of lactate dehydrogenase immobilized on "silica sol gel/nanometre-sized tridecameric aluminum polycation" modified gold electrode and its application. AB - This paper reports the electrochemical behavior of lactate dehydrogenase (LDH) immobilized in the silica sol-gel film on gold electrode after adding nanometre sized tridecameric aluminium polycation (nano-Al13, also called nanopolynuclear Al13) as a promoter. A pair of surface controlled quasi-reversible cyclic voltammetry peaks with the formal potential (E0') of 154 mV (vs. SCE) was found in the presence of nano-Al13. A potential application of the nano-Al13-LDH electrode for the determination of resorcinol and p-xylene was also investigated. The experimental results showed that both resorcinol and p-xylene inhibited LDH activity, and the calibration ranges were 5.0 x 10(-6)-3.0 x 10(-4) mol L(-1) for resorcinol and 1.0 x 10(-6)-1.0 x 10(-5) mol L(-1) for p-xylene, respectively. The nano-Al13-LDH electrode can be anticipated to be applied to environmental toxic assessments. PMID- 19562208 TI - The investigation of photo-induced chemiluminescence on Co2+-doped TiO2 nanoparticles and its analytical application. AB - A novel space- and time-resolved photo-induced chemiluminescence (PICL) analytical method was developed based on the photocatalysis of the Co2+-doped TiO2 nanoparticles. The PICL reaction procedure under the photocatalysis of Co2+ doped TiO2 nanoparticles was investigated using cyclic voltammetry and potentiometry. Meanwhile, the effect of the electrical double layer outside the Co2+-doped TiO2 nanoparticles on the PICL was investigated by contrasting with the Co2+-doped TiO2-SiO2 core-shell nanoparticles. Significantly, the CL intensity increased apparently and the time of the CL was prolonged in the presence of procaterol hydrochloride because the mechanism of the enhanced PICL reaction may be modified. The route of the PICL was changed due to the participation of the procaterol hydrochloride enriched at the surface of the Co2+ doped TiO2-SiO2 in the PICL reaction, which prolonged the time of the CL reaction and resulted in the long-term PICL. The analytical characteristics of the proposed in-situ PICL method were investigated using the procaterol hydrochloride as the model analyte. The investigation results showed that this new PICL analytical method offered higher sensitivity to the analysis of the procaterol hydrochloride and the PICL intensity was linear with the concentration of the procaterol hydrochloride in the range from ca. 2.0 x 10(-10) to 1.0 x 10(-8) g mL(-1). PMID- 19562209 TI - Development of robust quantitative methods by near-infrared spectroscopy for rapid pharmaceutical determination of content uniformity in complex tablet matrix. AB - Robust NIR transmission spectroscopic methods have been developed for determination of content uniformity (CU) of pharmaceutical products with a complex tablet matrix. The tablets of interest, formulated with eight components with active drug load of approximately 30% (w/w), are non-film coated, embossed, and round with thickness values of 3.6 and 5.6 mm, for the 125 and 500 mg dosage strength, respectively. The calibration data set contained seven laboratory scale batches of tablets with concentration range of active pharmaceutical ingredients (API) varying from 85 to 115% relative to label claim (LC) as well as four full scale production batches of tablets that included the natural physical variability of tablets. The reference concentration values were established by a high performance liquid chromatographic method. Partial least-squares (PLS) regression method was used to generate the calibration models. The root mean squared error of calibration for 125 and 500 mg was 1.6 and 1.5% in LC, respectively. The calibration models were validated in terms of measurement accuracy, repeatability, precision, robustness and transferability. Robustness assessment involved challenging the model with tablets incorporating variations in hardness, excipient vendors, excipient content and excipient particle size. The methods exhibited excellent measurement accuracy based on 87 batches (ten tablets for each batch) evaluated. The transferability of the developed NIR methods was demonstrated by comparing the NIR CU results associated with the same set of tablets scanned at the development site with those scanned at the production site. The result indicates that the NIR method can be used as a suitable alternative to the HPLC method for rapid tablet CU release test in pharmaceuticals. PMID- 19562210 TI - Ultrasound-enhanced enzymatic hydrolysis of conjugated female steroids as pretreatment for their analysis by LC-MS/MS in urine. AB - A fast, selective and sensitive method is here proposed for the analysis of female steroid hormones as conjugated forms (mainly, glucuronides and sulfates). The method has been applied to female urine samples to assess the metabolism of these compounds. The method implements an enzymatic hydrolysis (beta glucuronidase with sulfatase activity) kinetically enhanced by ultrasonic energy in order to generate the free steroid forms. This enables a drastic shortening of the time required for this step as compared with conventional protocols (from 12 18 hours to 30 min). The reaction kinetics of the ultrasound-enhanced hydrolysis was characterized by comparison to that of the conventional protocol. After hydrolysis, the free steroid hormones were isolated and preconcentrated by automated solid-phase extraction and the eluate was subsequently analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The target analytes were confirmed and quantified by multiple reaction monitoring (MRM). The detection and quantification limits were within 0.06-0.8 ng mL(-1) and 0.19-2.69 ng mL(-1), respectively. The precision of the method, expressed as intra-day and inter-day variability, ranged between 2.1 and 5.2% and between 4.9 and 8.0%, respectively. A complementary study was carried out to assess the storage conditions of urine samples. This study is crucial in those applications involving metabolic processes as the integrity of the sample has to be preserved. PMID- 19562211 TI - Disposable electrochemiluminescent biosensor for lactate determination in saliva. AB - An electrochemiluminescence-based disposable biosensor for lactate is characterized. The lactate recognition system is based on lactate oxidase (LOx) and the transduction system consists of luminol. All the needed reagents, luminol, LOx, BSA, electrolyte and buffer have been immobilized by a Methocel membrane placed on the working electrode of the screen-printed electrochemical cell. The measurement of the electrochemiluminescence (ECL) is made possible via a photocounting head when 50 microl of sample is placed into the screen-printed cell with a circular container containing the disposable sensing membrane. The compositions of the membrane and reaction conditions have been optimized to obtain adequate sensitivity. The disposable biosensor responds to lactate after 20 s when two 1 s pulses at 0.5 V are applied to obtain the analytical parameter, the ECL initial rate. The linearized double logarithmic dependence for lactate shows a dynamic range from 10(-5) to 5 x 10(-4) M with a detection limit of 5 x 10(-6) M and a sensor-to-sensor repeatability, as relative standard deviation, RSD, of 3.30% at the medium level of the range. The ECL disposable biosensor was applied to the analysis of lactate in human saliva as an alternative procedure for obtaining the lactate level in a non-invasive way. Interferences coming from components of saliva were studied and eliminated in a simple way that was easy to handle. The procedure was validated for use in human saliva, comparing the results against an enzymatic reference procedure. The proposed method is quick, inexpensive, selective and sensitive and uses conventional ECL instrumentation. PMID- 19562212 TI - A regenerable immunochip for the rapid determination of 13 different antibiotics in raw milk. AB - Access to high-quality and safe food is a basic need in our community and, consequently, the European Union has defined maximum residue levels (MRLs) for a number of antibacterial compounds. However, despite the obvious demand for quantitative multi-residue detection methods that can be carried out on a routine basis, there is currently a lack in the development of such systems. In particular, an automated multianalyte detection instrument is needed that is capable of quantifying several antibiotics simultaneously within minutes. The newly developed hapten microarrays are designed for the parallel analysis of 13 different antibiotics in milk within six minutes by applying an indirect competitive chemiluminescence microarray immunoassay (CL-MIA). To allow multiple analyses, a regenerable microarray chip was developed based on epoxy-activated PEG chip surfaces, onto which microspotted antibiotic derivatives like sulfonamides, beta-lactams, aminoglycosides, fluorquinolones and polyketides are coupled directly without further use of linking agents. Using the chip reader platform MCR 3, this antigen solid phase is stable for at least 50 consecutive analyses. PMID- 19562213 TI - Detection of drug-induced cellular changes using confocal Raman spectroscopy on patterned single-cell biosensors. AB - We report on a cell-based biosensor application that utilizes patterned single cell arrays combined with confocal Raman spectroscopy to observe the time dependent drug response of individual cells in real time. The patterned single cell platform enables individual cells to be easily located and continuously addressable for Raman spectroscopy characterization of biochemical compositional changes in a non-destructive, quantitative manner so that discrete cellular behavior and cell-to-cell variations are preserved. In this study, human medulloblastoma (DAOY) cells were exposed to the common chemotherapeutic agent etoposide, and Raman spectra from patterned cells were recorded over 48 hours. It was found that 87.5% of the cells monitored exhibited a sharp decrease in DNA and protein associated peaks 48 hours after drug exposure, corresponding to cell death. The remaining 12.5% of the cells showed little to no reduction in key Raman biomarkers, indicating their drug resistance. Furthermore, the patterned cell population showed a very similar response to etoposide as confluent cell cultures, as confirmed by flow cytometry. Finally, patterned cells were assessed with TUNEL assay for apoptosis due to DNA fragmentation after etoposide exposure. The results agree well with those from the Raman spectroscopy analysis. This combined biosensor-Raman platform provides a quick, simple way to assess cell responses to chemical and biological agents with high throughput and can be potentially used for a wide variety of biomedical applications such as pharmaceutical drug discovery, toxin tests, and biothreat detection. PMID- 19562214 TI - Probing inhibitory effects of destruxins from Metarhizium anisopliae using insect cell based impedance spectroscopy: inhibition vs chemical structure. AB - A noninvasive technique based on electric cell-substrate impedance sensing (ECIS) was demonstrated for on-line probing inhibitory effects of five destruxins on Spodoptera frugiperda Sf9 insect cells. Such chemically structurally similar cyclic hexadepsipeptides, were isolated and purified from the fungus Metarhizium anisopliae. Based on a response function, the inhibitory effect of the destruxins was established from determining the half-inhibition concentration (ECIS50), i.e., the level at which 50% inhibition of the cell response was obtained. Probing by cell based impedance spectroscopy indicated that only a slight change in their chemical structures provoked a significant effect on inhibition. Destruxin B was most inhibitory but replacement of a single methyl group with hydrogen (destruxin B2) or addition of a hydroxyl group (destruxin C) significantly reduced the inhibition. The removal of one methyl group and one hydrogen (destruxin A) lowered the inhibitory effect even more whereas the formation of an epoxy ring (destruxin E) in the structure nullified the inhibitory effect. PMID- 19562215 TI - Selective UV-filter detection with sensors based on stainless steel electrodes modified with polyaniline doped with metal tetrasulfonated phthalocyanine films. AB - This work describes the construction and application of two amperometric sensors for sensitive UV-filter determination. The sensors were prepared using stainless steel electrodes in which polyaniline (PANI) was electrochemically polymerized in the presence of nickel (NiPcTS) or iron (FePcTS) tetrasulfonated phthalocyanines. The sensor surface characterizations were carried out using atomic force microscopy (AFM). The PANI/NiPcTS sensor was selective for the chemical UV-filter p-aminobenzoic acid (PABA) and the PANI/FePcTS sensor was selective for octyldimethyl-PABA (ODP), both in a mixture of tetrahydrofuran (THF) and 0.1 mol L(-1) H2SO4 at a volume ratio of 30 : 70, and with an applied potential of 0.0 mV vs. Ag|AgCl. A detailed investigation of the selectivity was carried out for both sensors, in order to determine their responses for ten different UV filters. Finally, each sensor was successfully applied to PABA or ODP quantification in sunscreen formulations and water from swimming pools. PMID- 19562216 TI - Viral infection of cells in culture detected using infrared microscopy. AB - FTIR microscopy has been used to collect spectra for uninfected (mock) Vero cells, and cells that have been infected with herpes simplex virus type 1 (HSV-1) and human adenovirus type 5 (Ad-5). Cells were infected at a multiplicity of infection of 10, and studied at 24 hours post exposure. The spectra for infected samples display many differences compared to the spectra for uninfected samples. To estimate how well the spectra for uninfected and infected samples could be discriminated, we used logistic and partial least squares regression methods. We show that the spectra for HSV-1 and mock infected samples are well differentiated and, for a sensitivity of 95%, we calculate a specificity of 0.999 using partial least squares regression. Spectra for Ad-5 and mock infected samples are also well differentiated. We find that applying our regression models constructed with one data set to a new validating data set still gives very high levels of specificity for a given sensitivity. Spectra for Ad-5 and HSV-1 infected samples are also differentiable. Applying our constructed regression models to new validating data, however, leads to a decrease in the discrimination capability in this instance. If one is simply interested in differentiating spectra associated with uninfected and infected cells, without distinguishing the type of infection, then we show that logistic regression models can break down whereas partial least squares regression models perform well. PMID- 19562217 TI - Selectivity enhancement in neutral-carrier-type ion sensors based on a liquid crystalline polymer. AB - A liquid-crystalline polymer was applied to a neutral-carrier-type ion sensor as a membrane material for enhancement of ion selectivity. An ion-sensitive field effect transistor (ISFET) based on the membranes containing the polymer showed high sensitivity and selectivity for K+ by adding a low-molecular-weight liquid crystal as the plasticizer. The ion-sensor properties of the ISFETs based on the membranes containing a liquid-crystalline polymer were similar to those of the corresponding ion-selective electrodes (ISEs) based on the same polymer. The ion sensor with a liquid-crystalline polymer was more durable than that without any liquid-crystalline polymer. The use of liquid-crystalline polymer as the membrane material improves not only the ion-sensor properties but also the durability. PMID- 19562218 TI - Turn-on fluorescent detection of cyanide based on the inner filter effect of silver nanoparticles. AB - A simple, sensitive fluorescent method for detecting cyanide has been developed based on the inner filter effect (IFE) of silver nanoparticles (Ag NPs). With a high extinction coefficient and tunable plasmon absorption feature, Ag NPs are expected to be a powerful absorber to tune the emission of the fluorophore in the IFE-based fluorescent assays. In the present work, we developed a turn-on fluorescent assay for cyanide based on the strong absorption of Ag NPs to both excitation and emission light of an isolated fluorescence indicator. In the presence of cyanide, the absorber Ag NPs will dissolve gradually, which then leads to recovery of the IFE-decreased emission of the fluorophore. The concentration of Ag NPs in the detection system was found to affect the fluorescence response toward cyanide greatly. Under the optimum conditions, the present IFE-based approach can detect cyanide ranging from 5.0 x 10(-7) to 6.0 x 10(-4) M with a detection limit of 2.5 x 10(-7) M, which is much lower than the corresponding absorbance-based approach and compares favorably with other reported fluorescent methods. In addition, the present method possesses a good selectivity for cyanide over other common anions and further application in cyanide-spiked water samples suggested a recovery between 98.2 and 101.4%. Therefore, our proposed IFE-based fluorescent method is expected to be applied for cyanide determination in practical applications. PMID- 19562220 TI - New technology for in situ visualization, monitoring and controlling microwave chemical reaction progress using a focused microwave high pressure-temperature closed-vessel digestion system. AB - An experimental model has been developed for studying microwave energy transfer to a sample/solution placed in a high-pressure closed-vessel focused microwave heated digestion system. The absorption of microwave radiation with a frequency of 2.45 GHz in sample solutions was studied as a function of the efficiency of sample decomposition by measuring the incident and reflected power, temperature and pressure. Experimental measurements of incident/reflected power were carried out with a microwave detector for one type of reagent classically used in wet digestion techniques, namely nitric acid. HNO3 had been chosen as a digesting reagent to reduce the complexity of the observations to a single power-absorbing component. Results obtained by examining pressure, temperature, incident and reflected power graphs were compared with treatment efficiencies determined by measuring total organic carbon variation, arsenobetaine and arsenic content as well as trace element (Cu, Fe, Mn, Zn) recovery in the digests. The reflected power of a sample after microwave-assisted digestion was successfully used as an indicator to evaluate the digestion efficiency. Methodology was developed using the powdered biological certified reference material (DORM-2). PMID- 19562219 TI - A methodology for quantitation and characterization of oligonucleotides in albumin microspheres. AB - A fluorescence assay was developed to quantify oligonucleotides (ODNs) encapsulated in bovine serum albumin (BSA) microspheres using antisense to Nuclear Factor-kappaB (NF-kappaB) as a model ODN and employing Oligreen as the fluorescent dye. Methodologies were optimized for the suspension of the microspheres as well as release of the encapsulated ODN using protease digestion. This was followed by the detection and quantitation of the ODN using the Oligreen dye. The Oligreen fluorescence assay gave a concentration-dependent fluorescent interaction with the ODN. Further characterization of the ODN with respect to their structural integrity in non-irradiated and gamma-irradiated antisense encapsulated in BSA microspheres was performed using HPLC, infrared spectroscopy and polyacrylamide gel electrophoresis. Results showed no structural modification of antisense in the BSA microspheres as determined by HPLC retention times for the pure antisense and microsphere-encapsulated ODN. The migration pattern of the antisense in polyacrylamide gels confirmed the absence of significant alterations as a result of the encapsulation process or due to gamma-irradiation. The infrared spectra of non-irradiated and gamma-irradiated antisense to NF-kappaB microspheres also displayed peaks characteristic of the functional groups. The fluorescence assay could also detect NF-kappaB antisense in the serum of rats administered with encapsulated antisense by oral and intravenous routes. This methodology should be valuable for the analysis of BSA-encapsulated antisense ODN and for pharmacokinetic studies during antisense therapy. PMID- 19562221 TI - Two-parameter monitoring in a lab-on-valve manifold, applied to intracellular H2O2 measurements. AB - This work introduces, for the first time, simultaneous monitoring of fluorescence and absorbance using Bead Injection in a Lab-on-valve format. The aim of the paper is to show that when the target species, cells immobilized on a stationary phase, are exposed to reagents under well-controlled reaction conditions, dual monitoring yields valuable information. The applicability of this technique is demonstrated by the development of a Bead Injection method for automated measurement of cell density and intracellular hydrogen peroxide. PMID- 19562229 TI - Intuitive and axiomatic arguments for quantifying diagnostic test performance in units of information. AB - OBJECTIVES: Mutual information is a fundamental concept of information theory that quantifies the expected value of the amount of information that diagnostic testing provides about a patient's disease state. The purpose of this report is to provide both intuitive and axiomatic descriptions of mutual information and, thereby, promote the use of this statistic as a measure of diagnostic test performance. METHODS: We derive the mathematical expression for mutual information from the intuitive assumption that diagnostic information is the average amount that diagnostic testing reduces our surprise upon ultimately learning a patient's diagnosis. This concept is formalized by defining "surprise" as the surprisal, a function that quantifies the unlikelihood of an event. Mutual information is also shown to be the only function that conforms to a set of axioms which are reasonable requirements of a measure of diagnostic information. These axioms are related to the axioms of information theory used to derive the expression for entropy. RESULTS: Both approaches to defining mutual information lead to the known relationship that mutual information is equal to the pretest uncertainty of the disease state minus the expected value of the posttest uncertainty of the disease state. Mutual information also has the property of being additive when a test provides information about independent health problems. CONCLUSION: Mutual information is the best single measure of the ability of a diagnostic test to discriminate among the possible disease states. PMID- 19562228 TI - Computation of a probabilistic statistical shape model in a maximum-a-posteriori framework. AB - OBJECTIVES: When analyzing shapes and shape variabilities, the first step is bringing those shapes into correspondence. This is a fundamental problem even when solved by manually determining exact correspondences such as landmarks. We developed a method to represent a mean shape and a variability model for a training data set based on probabilistic correspondence computed between the observations. METHODS: First, the observations are matched on each other with an affine transformation found by the Expectation-Maximization Iterative-Closest Points (EM-ICP) registration. We then propose a maximum-a-posteriori (MAP) framework in order to compute the statistical shape model (SSM) parameters which result in an optimal adaptation of the model to the observations. The optimization of the MAP explanation is realized with respect to the observation parameters and the generative model parameters in a global criterion and leads to very efficient and closed-form solutions for (almost) all parameters. RESULTS: We compared our probabilistic SSM to a SSM based on one-to-one correspondences and the PCA (classical SSM). Experiments on synthetic data served to test the performances on non-convex shapes (15 training shapes) which have proved difficult in terms of proper correspondence determination. We then computed the SSMs for real putamen data (21 training shapes). The evaluation was done by measuring the generalization ability as well as the specificity of both SSMs and showed that especially shape detail differences are better modeled by the probabilistic SSM (Hausdorff distance in generalization ability Re approximately 25% smaller). CONCLUSIONS: The experimental outcome shows the efficiency and advantages of the new approach as the probabilistic SSM performs better in modeling shape details and differences. PMID- 19562230 TI - Computer-assisted diagnosis for precancerous lesions in the esophagus. AB - OBJECTIVES: The interpretation of endoscopic findings by gastroenterologists is still a difficult and highly subjective task. Despite important developments such as chromo-endoscopy, pit pattern analysis, fluorescence imaging as well as narrow band imaging it still requires lots of experience and training with a certain tentativeness until the final biopsy. By the development of computer-assisted diagnosis (CAD) systems this process can be supported. METHODS: This paper presents a new approach to CAD for precancerous lesions in the esophagus based on color-texture analysis in a content-based image retrieval (CBIR) framework. The novelty of our approach lies in the combination of newly developed color-texture features with the interactive feedback loop provided by a relevance feedback algorithm. This allows the expert to steer the query and is still robust against accidental false decisions. RESULTS: We reached an inter-rater reliability of kappa = 0.71 on a database of 390 endoscopic images. The retrieval accuracy didn't change significantly until a wrong decision rate of 20%. CONCLUSIONS: Thus, the system could be able to support practitioners with less experience or in private practice. In combination with a connected case database it can also support case-based reasoning for the diagnostic decision process. PMID- 19562231 TI - Impact of vaccination against varicella on the reduction of the disease incidence in children and adolescents from Florianopolis, Brazil. AB - OBJECTIVE: To evaluate the impact of universal early childhood vaccination against varicella on the reduction of its incidence in a state capital in southern Brazil. METHODS: Official epidemiologic surveillance data from the SINAN database for the 1997-2007 period were used to evaluate the impact of varicella vaccination targeting all children < 2 years of age in Florianopolis (the capital city of the state of Santa Catarina), Brazil, since 2002, thus comprising 5 years before and 6 years after the vaccination. Varicella incidence in Florianopolis was compared with the incidence in the rest of the state for four age groups (< 1, 1-4, 5-9, and 10-14 years). RESULTS: Among the 135,311 cases of varicella in the state of Santa Catarina during the 1997-2007 period, 70% were children under 10 years of age. The effectiveness of varicella vaccine ranged from 27 to 38% among the age groups but reached statistical significance only for children 1-4 years old. CONCLUSION: The vaccine was effective in reducing varicella incidence in Florianopolis for children 1-4 years old. PMID- 19562232 TI - Women, patients with severe asthma, and patients attended by primary care physicians, are at higher risk of suffering from poorly controlled asthma. AB - AIM: The aim of this exploratory study was to identify those factors associated with asthma control assessed according to GINA Guidelines. METHOD: 809 (56% female) subjects with asthma were recruited consecutively from both specialist and primary care centres. Asthma control was assessed over a 4-week follow-up period using a composite measure. A multivariate analysis was performed, in which asthma control was included as the dependent variable and several explanatory variables were included as independent variables. RESULTS: Analysis performed on the whole population rendered gender (p=0.003), the type of physician (p<0.001), and age group (p<0.001), as significant factors associated with asthma control. In adults, gender (p=0.001), asthma severity (p<0.001), and type of physician (p<0.001) were significant, and only asthma severity was significant (p=0.043) in children. CONCLUSIONS: After model adjustment, we suggest that being female, suffering from more severe asthma, or being attended by a primary care physician, could pose a significantly higher risk of having poorly controlled asthma in adults. PMID- 19562233 TI - The value of self-report assessment of adherence, rhinitis and smoking in relation to asthma control. AB - AIMS: To explore the utility of self-report measures of inhaled corticosteroid (ICS) adherence, degree of rhinitis and smoking status and their association with asthma control. METHODS: Patients prescribed ICS for asthma at 85 UK practices were sent validated questionnaire measures of control (Asthma Control Questionnaire; ACQ) and adherence (Medication Adherence Report Scale), a two-item measure of smoking status, and a single-item measure of rhinitis. RESULTS: Complete anonymised questionnaires were available for 3916 participants. Poor asthma control (ACQ >1.5) was associated with reported rhinitis (OR = 4.62; 95% CI: 3.71-5.77), smoking (OR = 4.33; 95% CI: 3.58-5.23) and low adherence to ICS (OR = 1.35; 95% CI: 1.18-1.55). The degree of rhinitis was important, with those reporting severe rhinitis exhibiting the worst asthma control, followed by those reporting mild rhinitis and then those reporting no rhinitis symptoms (F(2, 3913)=128.7, p<.001). There was a relationship between the number of cigarettes smoked each day and asthma control (F(5,655)=6.08, p<.001). CONCLUSIONS: Poor asthma control is associated with self-reported rhinitis, smoking and low medication adherence. These potentially modifiable predictors of poor asthma control can be identified through a brief self-report questionnaire, used routinely as part of an asthma review. PMID- 19562234 TI - The importance of the psychosocial work environment for employee well-being and work motivation. PMID- 19562236 TI - Common prognostic factors of work disability among employees with a chronic somatic disease: a systematic review of cohort studies. AB - OBJECTIVE: Based on prospective and retrospective disease cohort studies, the aim of this review was to determine common prognostic factors for work disability among employees with rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, diabetes mellitus, and ischemic heart disease (IHD). METHODS: A systematic literature search in Medline (1990-2008) and Embase (1990-2008) was carried out to identify relevant cohort studies using a well-defined list of inclusion and quality criteria. RESULTS: We identified 43 relevant cohort studies with sufficient methodological quality (20 for rheumatoid arthritis, 3 for asthma and 20 for IHD). The common prognostic factors for work disability found in all the diseases were: perceived health complaints, limitation in daily physical activities caused by the disease (high versus low), heavy manual work, and female gender. The common positive prognostic factors for rheumatoid arthritis and IHD were age (high versus low) and sickness absence. The common negative factors for rheumatoid arthritis and IHD were education (high versus low) and ethnic origin (white versus non-white). CONCLUSIONS: As many prognostic factors for work disability are similar for employees with various chronic diseases, it is possible to detect high risk groups. This information supports the development and implementation of a general disability management intervention for employees suffering from a chronic disease to overcome health-related limitations at work. PMID- 19562235 TI - Validity of self-reported mechanical demands for occupational epidemiologic research of musculoskeletal disorders. AB - OBJECTIVES: To describe the relation of the measured validity of self-reported mechanical demands (self-reports) with the quality of validity assessments and the variability of the assessed exposure in the study population. METHODS: We searched for original articles, published between 1990 and 2008, reporting the validity of self-reports in three major databases: EBSCOhost, Web of Science and PubMed. Identified assessments were classified by methodological characteristics (eg, type of self-report and reference method) and exposure dimension was measured. We also classified assessments by the degree of comparability between the self-report and the employed reference method, and the variability of the assessed exposure in the study population. Finally, we examined the association of the published validity (r) with this degree of comparability, as well as with the variability of the exposure variable in the study population. RESULTS: Of the 490 assessments identified, 75% used observation-based reference measures and 55% tested self-reports of posture duration and movement frequency. Frequently, validity studies did not report demographic information (eg, education, age, and gender distribution). Among assessments reporting correlations as measure of validity, studies with a better match between the self-report and the reference method, and studies conducted in more heterogeneous populations tended to report higher correlations [odds ratio (OR) 2.03, 95% confidence interval (95% CI) 0.89 4.65 and OR 1.60, 95% CI 0.96-2.61, respectively]. CONCLUSIONS: The reported data support the hypothesis that validity depends on study-specific factors often not examined. Experimentally manipulating the testing setting could lead to a better understanding of the capabilities and limitations of self-reported information. PMID- 19562237 TI - [Laparoscopy for suspected appendicitis. Should an appendix that appears normal be removed?]. AB - The question whether an appendix found to be macroscopically normal at laparoscopy for suspected appendicitis should be removed remains open to debate. Potential advantages of appendicectomy in all cases include early diagnosis of neoplastic lesions that cannot be detected macroscopically, diagnosis and cure of neurogenic appendicectomy, avoidance of diagnostic confusion in later episodes of abdominal pain, and prevention of appendicitis developing later in life. Therefore, adopting a strategy of always removing the appendix even if it is found to be uninflamed at laparoscopy seems justified as long as it does not imply an increase in postoperative morbidity. We retrospectively studied all patients undergoing laparoscopic appendicectomy in which a "normal appendix" was found and all patients undergoing diagnostic laparoscopy in our hospital during a 7-year period. Our data as well as a critical review of the literature show that removal of the appendix does not increase morbidity compared to simple diagnostic laparoscopy and should always be done when performing laparoscopy for suspected acute appendicitis. PMID- 19562238 TI - [Acute appendicitis: current surgical value of diagnostics and therapy standards]. PMID- 19562239 TI - [Appendicitis in the elderly. CRP value as decision support for diagnostic laparoscopy]. AB - BACKGROUND: The diagnosis of acute appendicitis in the elderly bears many pitfalls due to a broad range of differential diagnoses and uncommon clinical presentation. This may result in late detection of appendicitis leading to poor outcome. The aim of this study was to examine the characteristics of appendicitis in elderly patients in order to facilitate early diagnosis. MATERIALS AND METHODS: All patients who underwent appendectomy in our institution were prospectively recorded over a 30 month period. Data on patient's age (>60 years versus < or =60 years), clinical findings, the inflammatory parameters leucocytes and C-reactive protein (CRP) and histological-findings (perforated versus non perforated) were collected. Statistical analysis was carried out by ROC analysis, chi(2) and t-tests. RESULTS: In the examination period 403 patients underwent appendectomy and 11.2% (n=45) were older than 60 years. These patients were characterized by significantly more frequent perforations compared to those patients < or =60 years (35.6% versus 7.0%, p< or =0.05), peritonitis (42.2% versus 9.5%, p< or =0.05), conversion to open surgery (23% versus 5%, p< or =0.005), longer postoperative hospital stay (9.2 days versus 4.3 days, p< or =0.05) and a higher complication rate (28.9% versus 3.6%, p< or =0.005). CRP values in patients >60 years were on average 123.2 mg/l and significantly higher than in patients < or =60 years (35.5 mg/l, p< or =0.005). The ROC analysis resulted in a CRP cut-off value of 101.9 mg/l for patients >60 years for the existence of a perforation with a specificity of 72.4% and a sensitivity of 81.3% (AUC 0.811). CONCLUSIONS: The CRP value showed a strong correlation with respect to the grade of inflammation and perforation. In conclusion, elderly patients with symptoms of appendicitis and a CRP value higher than 102 mg/l should undergo early diagnostic laparoscopy. PMID- 19562240 TI - [PET-CT studies of metastasizing cancer of the colon and rectum. Variability of tumor aggressiveness as a micro-evolutionary process of cancer stem cells with predetermined prognosis]. AB - Formation of cancer stem cells which are both rare and variably therapy-resistant marks the beginning of a new disease without precursors. Based on molecular changes, these cells are derived from normal cells and exhibit pre-programmed malignant behaviour. In vitro studies have shown that hybrid cancers which behave in a similar way to Dukes A, B or C cancers in vivo can be produce by horizontal gene transfer. The level of aggressiveness follows a Galton curve in the probability distribution. In the current paper we analyzed colorectal cancers by PET-CT in follow-up studies which extended over several years. We conclude that the primary tumors behave differently from distant metastases. Radical exstirpation of the primary tumor is able to cure the malignant process if the homing area is resected. The primary tumor acts as the supplier of cancer stem cells for metastases which appear in different organs. When chemotherapy is administered the distribution of metastases in different organs appears dependent of the response or non-response of cancer stem cells to this therapy. Large numbers of colorectal carcinomas existed for the same time duration before death (15 years) independent of the malignancy grade. The tumor metastasizes immediately after formation. The primary tumor and the metastases appear variably quickly depending on the malignancy grade and are autonomic processes. PMID- 19562241 TI - Alendronate-related femoral diaphysis fracture--what should be done to predict and prevent subsequent fracture of the contralateral side? AB - INTRODUCTION: Atypical fractures of the femoral diaphysis have recently been associated with alendronate therapy (Neviaser et al. J Orthop Trauma 22(5):346 350, 2008; Kwek et al. Injury 39:224-231, 2008; Lenart et al. N Engl J Med 358:1304-1306, 2008). METHODS: In many cases, fractures have occurred bilaterally prompting debate regarding appropriate screening of the unaffected side (Kwek et al. N Engl J Med 359(3):316-317, 2008). CASE REPORT: We report a case of sequential, bilateral, femoral diaphysis fractures associated with prolonged alendronate therapy and the failure to predict the subsequent fracture of the contralateral side despite radiological imaging. DISCUSSION: We review the current literature and discuss potential management strategies. PMID- 19562242 TI - Age-related changes in biochemical markers of bone turnover and gonadotropin levels and their relationship among Chinese adult women. AB - The relationship between the levels of gonadotropic hormones and bone metabolism in Chinese adult women is unclear. Our research shows that a significant positive correlation exists between the levels of gonadotropic hormones and various bone turnover indicators. Follicle-stimulating hormone (FSH) has been found to have a greater influence on all types of bone turnover indicator than luteinizing hormone (LH). Further, FSH has a greater influence on bone formation indicators than on bone resorption indicators. INTRODUCTION: The aim of this study was to investigate the relationship between serum FSH and LH and biochemical markers of bone turnover in native Chinese adult women. METHODS: We conducted a cross sectional study of 694 healthy Chinese women aged between 20 and 82 years. Serum FSH, LH, bone-specific alkaline phosphatase (BAP), osteocalcin (OC), N-terminal telopeptides of type I collagen, C-terminal telopeptides of type I collagen, urinary NTX, urinary CTX, and urinary deoxypyridinoline (uDPD) were determined. RESULTS: All types of bone turnover indicator were significantly positively correlated with FSH (r = 0.164-0.626, all P = 0.000) and LH (r = 0.130-0.618, all P = 0.013-0.000). The correlation coefficient between serum FSH and BAP was the highest (r = 0.626), and that between serum FSH and uDPD was the lowest (r = 0.164). The serum gonadotropic hormone levels were higher; concentrations of bone turnover indicators were higher. The extent of the influence of FSH on various bone turnover indicators was approximately seven to 20 times greater than that of LH on these indicators. FSH could explain 43% and 22% of the changes in BAP and OC, respectively; whereas, LH could explain only 2.1% and 1.1%, respectively. FSH could explain approximately 1.9-11.8% of the changes in bone resorption indicators; however, LH had almost no effect on them. CONCLUSIONS: Gonadotropic hormone levels are correlated with the rate of bone turnover in Chinese women: the higher the serum gonadotropic hormone levels in circulation, the higher the levels of bone turnover indicators. FSH has a greater influence on all types of bone turnover indicator than LH; moreover, it has a greater influence on bone formation indicators than on bone resorption indicators. PMID- 19562243 TI - Progressive load training for the quadriceps muscle associated with proprioception exercises for the prevention of falls in postmenopausal women with osteoporosis: a randomized controlled trial. AB - This study aims to evaluate the effect of 18-week progressive muscular strength and proprioception training program on the muscle strength of the quadriceps, in prevention of falls in postmenopausal women with osteoporosis. The incidence of falls in the intervention group was significantly lower than in the control group (incidence rate ratio (IRR) = 0.263, 95% CI 0.10-0.68). INTRODUCTION: This study aims to evaluate the effect of a progressive muscular strength and proprioception training program on the muscle strength of the quadriceps, balance, quality of life, and reduction in the risk of falls in postmenopausal women with osteoporosis. METHODS: One hundred sedentary postmenopausal women with osteoporosis, ages ranging from 55 to 75, were selected and randomized into two groups: the intervention group comprised of 50 patients who underwent a 18-week of progressive load training for the quadriceps muscle (50% up to 80% of 1-RM-one maximum repetition) and proprioception training associated to a drug treatment of osteoporosis and the control group that included 50 patients who only underwent a drug treatment of osteoporosis. The muscular strength, balance, functional mobility, and quality of life were evaluated in the beginning and end of the research. The number of falls was evaluated 24 weeks post-treatment. RESULTS: Eighty-five patients concluded the research. The program promoted a significant difference among the groups for SF-36 in the eight sub-scales (p or = 2 adverse events were rash (59%), mucositis (34%) and diarrhea (31%). Grade 3 or higher events were rare. Best radiographic response included stable disease in 15 patients (47%); no patients achieved either a CR or PR. The estimated 6-month progression-free survival was 3.1% for all patients. Progression-free survival was better for patients not on EIAEDs (P = 0.03). Tumor markers failed to show an association with PFS except for increased pAKT expression which achieved borderline significance (P = 0.045). Although neither rash nor diarrhea had an association with outcome, hyperlipidemia was associated with longer PFS (P = 0.029). Erlotinib plus sirolimus was well tolerated but had negligible activity among unselected recurrent GBM patients. (ClinicalTrials.gov number: NCT0062243). PMID- 19562255 TI - Phase II trials of erlotinib or gefitinib in patients with recurrent meningioma. AB - There are no established treatments for recurrent meningioma when surgical and radiation options are exhausted. The epidermal growth factor receptor (EGFR) is often over-expressed in meningiomas and may promote tumor growth. In open label, single arm phase II studies of the EGFR inhibitors gefitinib (NABTC 00-01) and erlotinib (NABTC 01-03) for recurrent malignant gliomas, we included exploratory subsets of recurrent meningioma patients. We have pooled the data and report the results here. Patients with recurrent histologically confirmed meningiomas with no more than 2 previous chemotherapy regimens were treated with gefitinib 500 mg/day or erlotinib 150 mg/day until tumor progression or unacceptable toxicity. Twenty-five eligible patients were enrolled with median age 57 years (range 29 81) and median Karnofsky performance status (KPS) score 90 (range 60-100). Sixteen patients (64%) received gefitinib and 9 (36%) erlotinib. Eight patients (32%) had benign tumors, 9 (36%) atypical, and 8 (32%) malignant. For benign tumors, the 6-month progression-free survival (PFS6) was 25%, 12-month PFS (PFS12) 13%, 6-month overall survival (OS6) 63%, and 12-month OS (OS12) 50%. For atypical and malignant tumors, PFS6 was 29%, PFS12 18%, OS6 71%, and OS12 65%. The PFS and OS were not significantly different by histology. There were no objective imaging responses, but 8 patients (32%) maintained stable disease. Although treatment was well-tolerated, neither gefitinib nor erlotinib appear to have significant activity against recurrent meningioma. The role of EGFR inhibitors in meningiomas is unclear. Evaluation of multi-targeted inhibitors and EGFR inhibitors in combination with other targeted molecular agents may be warranted. PMID- 19562256 TI - Utility of administrative claims data for the study of brain metastases: a validation study. AB - In this study, we sought to determine the accuracy with which the International Classification of Diseases, 9th Edition, Clinical Modification (ICD-9-CM) diagnosis code for "secondary neoplasm of brain and spinal cord" in health insurance claims measures clinically evident central nervous system (CNS) metastases in patients with non-small cell lung cancer (NSCLC). For 241 consecutive patients with newly diagnosed NSCLC, we compared ICD-9-CM "secondary neoplasm" codes indicating tumor spread to the CNS from institutional billing records to gold-standard chart review to determine: (1) sensitivity, specificity and positive predictive value (PPV) of the site-specific secondary neoplasm code and (2) the accuracy in time of its appearance within billing records compared with the gold standard date of CNS relapse. The occurrence of at least one ICD-9 CM code for brain metastasis (Algorithm 1) had a sensitivity of 100% (95% CI: 100 100%) and PPV of 91% (95% CI: 87-94%). By requiring >or= 2 codes (Algorithm 2) or >or= 3 codes (Algorithm 3) for the diagnosis of brain metastasis in claims, specificity and PPV improved, while sensitivity did not drop substantially. The claims-based date of diagnosis was also accurate, with 92% of dates falling within 30 days of the gold standard. ICD-9-CM codes in institutional billing claims reliably documented NSCLC metastases to the CNS. These results suggest that Medicare claims data may be used to evaluate clinical and epidemiological issues related to brain metastases in elderly cancer patients. PMID- 19562257 TI - The Austrian Brain Tumour Registry: a cooperative way to establish a population based brain tumour registry. AB - In Austria, registration of malignant brain tumours is legally mandatory, whereas benign and borderline tumours are not reported. The Austrian Brain Tumour Registry (ABTR) was initiated under the auspices of the Austrian Society of Neuropathology for the registration of malignant and non-malignant brain tumours. All Austrian neuropathology units involved in brain tumour diagnostics contribute data on primary brain tumours. Non-microscopically verified cases are added by the Austrian National Cancer Registry to ensure a population-based dataset. In 2005, we registered a total of 1,688 newly diagnosed primary brain tumours in a population of 8.2 million inhabitants with an overall age-adjusted incidence rate of 18.1/100,000 person-years. Non-malignant cases constituted 866 cases (51.3%). The incidence rate was higher in females (18.6/100,000) as compared to males (17.8/100,000), while 95/1,688 (5.6%) cases were diagnosed in children (<18 years). The most common histology was meningioma (n = 504, 29.9%) followed by glioblastoma (n = 340, 20.1%) and pituitary adenoma (n = 151, 8.9%). Comparison with the Central Brain Tumor Registry of the United States (CBTRUS) database showed high congruency of findings. The ABTR model led by neuropathologists in collaboration with epidemiologists and the Austrian National Cancer Registry presents a cooperative way to establish a population-based brain tumour registry with high quality data. This setting links cancer registration to the mission of medical practice and research as defined by the World Medical Association in the Declaration of Helsinki. The continued operation of ABTR will aid in monitoring changes in incidence and in identifying regional disease clusters or geographic variations in brain tumour morbidity/mortality. PMID- 19562258 TI - WHO grade II and III meningiomas: a study of prognostic factors. AB - Meningiomas represent one of the largest subgroups of intracranial tumors. They are generally benign, but may show a histological progression to malignancy. Grades II and III meningiomas have been less well studied and are not well controlled because of their aggressive behaviour and recurrences. There is no consensus on therapeutic strategies and no prognostic factors are known. In order to determine these parameters, a multi-institutional retrospective analysis was performed in France with the support of the Neuro-Oncology Club of the French Neurosurgical Society. This study was performed on 199 adults treated for WHO grade II (166 patients) or grade III (33 patients) meningiomas between 1990 and 2004 in the Neurosurgery Departments of five French University Hospitals. Data on epidemiology, clinical behaviour and therapy were collected. Overall survival and progression-free survival were analysed as a function of each possible prognostic factor. For patients with grade II meningiomas, the 5- and 10-year OS rates were 78.4 and 53.3%, respectively, while, for patients with grade III meningiomas, the corresponding values were 44.0 and 14.2%. For patients with grade II meningiomas, the 5- and 10-year PFS rates were 48.4 and 22.6%, respectively, the corresponding values for patients with grade III meningiomas being 8.4 and 0%. For the grade II meningiomas, univariate analysis showed that age < 60 years (P < 0.0001) and Simpson 1 resection (P = 0.055) were associated with a longer OS. For the grade III meningiomas, univariate analysis showed that age < 60 years (P < 0.0001) and RT (P = 0.036) were associated with a longer OS. Histological grade II was found to be associated with a longer PFS (P = 0.0032) and RT reduced the PFS in grade II meningiomas (P = 0.0006) There were no other prognostic factors in terms of PFS for grades II and III meningiomas in univariate analysis. Multivariate analysis confirmed that age (< 60 years), Simpson 1 and histological grade II were independent prognostic factors for survival. This retrospective study might improve the management of grades II and III meningiomas. Prospective trials should delineate strong therapeutic guidelines for high-grade meningiomas. PMID- 19562259 TI - Human platelet alloantigens HPA-1, HPA-2, and HPA-3 polymorphisms associated with extent of severe coronary artery disease. AB - The contribution of human platelet antigen (HPA)-1 (GPIIb/IIIa), HPA-2 (GPIb/IX), and HPA-3 (GPIIb/IIIa) polymorphisms to the risk of coronary artery disease (CAD) was investigated in 341 CAD patients and 316 matched control subjects. HPA genotyping was performed by PCR-SSP. Regression analysis was employed in assessing the contribution of these variants to CAD risk. The frequency of HPA-1b (P = .009) and HPA-3b (P = .004) alleles, and HPA-1a/1b (P = .045), HPA-1b/1b (P = .007), and HPA-3b/3b (P = .008) genotypes were higher in patients than control subjects. No significant association was demonstrated between the HPA variants and 1-, 2- and 3-vessel disease. HPA-1b/2a/3b (Pc = .021) and HPA-1b/2b/3a (Pc = .002) haplotypes were positively associated with CAD, thereby conferring a disease susceptibility nature to these haplotypes. Multivariate analysis confirmed the positive association of HPA-1b/2a/3b (aOR = 3.72; 95% CI = 1.49 9.28), and in addition identified HPA-1b/2a/3a (aOR = 2.49; 95% CI = 1.06-5.86) to be positively associated with CAD, after adjusting for a number of covariates. Our results demonstrate positive association of HPA variants and specific HPA 1/HPA-2/HPA-3 haplotypes with CAD in Tunisians. PMID- 19562260 TI - Second-generation de novo design: a view from a medicinal chemist perspective. AB - For computational de novo design, a general retrospective validation work is a very challenging task. Here we propose a comprehensive workflow to de novo design driven by the needs of computational and medicinal chemists and, at the same time, we propose a general validation scheme for this technique. The study was conducted combining a suite of already published programs developed within the framework of the NovoBench project, which involved three different pharmaceutical companies and four groups of developers. Based on 188 PDB protein-ligand complexes with diverse functions, the study involved the ligand reconstruction by means of a fragment-based de-novo design approach. The structure-based de novo search engine FlexNovo showed in five out of eight total cases the ability to reconstruct native ligands and to rank them in four cases out of five within the first five candidates. The generated structures were ranked according to their synthetic accessibilities evaluated by the program SYLVIA. This investigation showed that the final candidate molecules have about the same synthetic complexity as the respective reference ligands. Furthermore, the plausibility of being true actives was assessed through literature searches. PMID- 19562261 TI - [Reconstruction of the acetabulum with structured bone graft in press-fit technique]. AB - OBJECTIVE: Reconstruction of defects of the superior acetabular rim with structured bone grafts in press-fit technique before total hip replacement. INDICATIONS: Defects of the superior acetabular rim following hip dysplasia Crowe type II-IV, avascular necrosis of the femoral head Ficat stage IV, or aseptic loosening of the cup with acetabular defects Paprosky type 2a and 2b. CONTRAINDICATIONS: Acetabular defects Paprosky type 2c, 3a and 3b, septic loosening, severe osteoporosis. SURGICAL TECHNIQUE: Exposure of the acetabular defect and debridement with a spherical reamer to create a concave bleeding graft bed. Shaping of the bone graft with an inverted reamer of corresponding size and oscillating saw. Press-fit insertion of the bone graft into the defect and temporary fixation with Kirschner wires. Rereaming of the acetabulum with the spherical reamer. Implantation of a cemented cup of corresponding size. Removal of Kirschner wires after setting of cement. POSTOPERATIVE MANAGEMENT: Full weight bearing in case of small or medium graft, 6-week non-weight bearing in case of big load-bearing graft. Mobilization on 1st day postoperatively. Stair climbing on 7th day postoperatively. Suture removal after 10 days. RESULTS: In 46 patients with 55 total hip replacements with structured grafts in press-fit technique, the Harris Hip Score improved from 38.9 points to 92.3 points after 29.4 months (12.0 84.4 months). There were two cases of delayed wound healing and one cup exchange because of aseptic loosening. Radiologically, one graft sintering by 5 mm was detected. PMID- 19562262 TI - ["Malignant" ARDS]. AB - Acute respiratory failure and the "acute respiratory distress syndrome" (ARDS) are frequent medical conditions in critically ill patients. Various causes can potentially result in the development of ARDS. Two cases are presented, in which malignant diseases were identified as causes of the respiratory failure. The first patient was diagnosed with an acute myeloic leukemia M5 (FAB). In the second patient, lung histology revealed an adenocarcinoma of the lung. These case reports show that in addition to the classical causes of ARDS, specific disease entities can mimic this form of respiratory failure. Beside solid cancers and lymphomas, acute and progressive forms of inflammatory, parenchymal lung diseases (such as acute interstitial pneumonitis, acute eosinophilic pneumonia, diffuse alveolar hemorrhagia, and acute hypersensitivity pneumonitis) can manifest with this picture. As a consequence, the diagnostic workup of respiratory failure of unknown cause should include these entities. PMID- 19562263 TI - [Cerebrovascular diseases]. AB - In the majority of the cases cerebrovascular disease is caused by atherosclerosis. Duplexsonography is the diagnostic tool of first choice. Management of cardiovascular risk factors is of paramount importance in secondary prevention of atherosclerotic vascular complications. Patients with a symptomatic internal carotid artery stenosis >70% have a clear indication for revascularization. Asymptomatic patients with >60% stenosis benefit from revascularisation if the perioperative risk for death or stroke is below 3%. The optimal revascularization strategy highly depends on the expertise of the local surgeon or endovascular specialist. In younger patients with cerebrovascular disease rare causes such as dissection, large vessel arteritis, fibromuscular dysplasia or vasospasms have to be considered. PMID- 19562264 TI - [Large vessel vasculitis]. AB - Large vessel vasculitis is characterized by local inflammation of the vessel wall of the aorta and its first and second degree branches. Giant cells are found regularly. The process leads to stenoses and occlusions as well as to aneurysmatic dilatation. Recent progress in vascular imaging (colour Doppler ultrasound, MRI and PET) allows to detect Takayasu arteritis at an earlier stage and to reveal the involvement of larger arteries in temporal arteritis (giant cell arteritis, GCA). With increasing frequency, a subtype of predominantly female and less aged GCA patients without cranial symptoms is noticed in whom obstructions are prevailing in the proximal arm vessels or in the aorta. The latter may be affected exclusively. Initial therapy is based on corticosteroids. In GCA, methotrexate hitherto is the only other immunosuppressive drug that has proved to be helpful. Disease activity has to be controlled both by laboratory parameters and vascular imaging. PMID- 19562265 TI - Biomechanical stability of an arthroscopic anterior capsular shift and suture anchor repair in anterior shoulder instability: a human cadaveric shoulder model. AB - It was hypothesized that an arthroscopic Bankart repair with suture anchors supplies sufficient anterior shoulder stability, which cannot be improved by an additional capsular shift. In an experimental biomechanical human cadaver study, we tested ten fresh human cadaver shoulders in a robot-assisted shoulder simulator. External rotation and glenohumeral translation were measured at 0 degrees and 80 degrees of glenohumeral abduction. All measurements were performed under the following conditions: on the non-operated shoulder; following the setting of three arthroscopic portals; following an arthroscopic anterior capsular shift; following a simulated Bankart lesion; and following an arthroscopic Bankart repair. The application of three arthroscopic portals resulted in a significant increase of the anterior (P = 0.01) and antero-inferior translation (P = 0.03) at 0 degrees and 80 degrees abduction, as well as an increase in external rotation at 80 degrees abduction (P = 0.03). Capsular shift reduced external rotation (P = 0.03), but did not significantly decrease translation. Simulating anterior shoulder instability, glenohumeral translation significantly increased, ranging from 50 to 279% of physiological translation. Arthroscopic shoulder stabilization resulted in a decrease of translation in all tested directions to approximately physiologic levels. External rotation in 0 degrees abduction was thus decreased significantly (P = 0.003) to an average of 19 degrees . The study proved that an arthroscopic anterior capsular shift in a cadaveric model decreases external rotation without a significant influence on glenohumeral translation. Arthroscopic shoulder stabilization with suture anchors thus sufficiently restores increased glenohumeral translation, but also decreases external rotation in neutral abduction. An anatomic reconstruction of the Bankart lesion without overconstraining of the antero-inferior capsule should therefore be the aim in arthroscopic anterior shoulder stabilization. PMID- 19562267 TI - Videofluorography swallow study of patients with systemic sclerosis. AB - PURPOSE: This study was undertaken to evaluate the role of the videofluorographic (VFG) swallow study in patients with systemic sclerosis. MATERIALS AND METHODS: Over a 23-month period, 45 women (mean age 58 years, range 27-76 years) with a known diagnosis of systemic sclerosis and a history of dysphagia underwent a dynamic and morphological study of the oral, pharyngeal and oesophageal phases of swallowing with videofluorography. All examinations were performed with a remote controlled digital C-arm device with 16-in image intensifier, 0.6- to 1.2-mm focal spot range and maximum tube voltage of 150 kVp in fluorography and 120 kVp in fluoroscopy. Cineradiographic sequences were acquired for the swallow study with 12 images per second and matrix 512 x 512 after the ingestion of boluses of high-density (250% weight/volume) barium. The evaluation of oesophageal peristalsis was documented with digital cineradiographic sequences with six images per second in the upright and supine positions during the swallowing of barium (60% weight/volume), and the water siphon test was performed with the patient in the supine position to evaluate the presence of gastro-oesophageal reflux disease (GORD). All patients subsequently underwent laryngoscopy, endoscopy and pH monitoring, and the data thus obtained were processed and compared. RESULTS: The VFG swallow study identified alterations of epiglottal tilting associated with intraswallowing laryngeal penetration in 26 patients (57.8%), pooling of contrast agent in the valleculae and pyriform sinuses in 23 (51.1%) and radiographic signs of nonspecific hypertrophy of the lingual and/or palatine tonsils in 18 (40%). The study of the oesophageal phase revealed the presence of altered peristalsis in all patients, and in particular, 36 patients (80%) showed signs of atony. Altered oesophageal clearing mechanisms were evident in all 45 patients, sliding hiatus hernia in 43 (93%) and GORD in 44 (97%). CONCLUSIONS: Our study demonstrated that in patients with systemic sclerosis, there is no primary alteration of the oral or pharyngeal phase of swallowing. In addition, alterations of epiglottal tilting associated with laryngeal penetration of contrast agent were found to be secondary to chronic GORD. Indeed, in 40% of patients, radiographic signs were found that indicated nonspecific hypertrophy of the lingual tonsil and/or palatine tonsils and nonspecific signs of chronic pharyngeal inflammation, and GORD was identified in 93% of patients, which in 40% of cases extended to the proximal third of the oesophagus. The data obtained were confirmed in 85% of cases with pH monitoring and in all cases with laryngoscopy. PMID- 19562266 TI - Magnetic resonance imaging of primary breast lymphoma. AB - PURPOSE: Primary lymphomas of the breast (PBNHL) are uncommon. Magnetic resonance imaging (MRI) features of these malignancies can be relevant in establishing the extent of disease and planning the appropriate therapeutic strategy, usually represented by chemo- and radiotherapy, rather than surgery. The purpose of this study was to assess MRI features of PBNHL. MATERIALS AND METHODS: MRI examinations performed on seven patients with known PBNHL were retrospectively evaluated. Lesions were analysed for both morphology and kinetics and classified according to the Breast Imaging Reporting and Data System (BI-RADS) categories. RESULTS: The mean MRI maximum diameter was 44 mm (range 12-69). Six lesions showed a mass-like enhancement; one lesion showed a non-mass-like enhancement. For mass-like lesions, kinetic curve assessment of initial rise showed slow enhancement in one lesion, rapid enhancement in four lesions and medium enhancement in one lesion. Assessment of delayed enhancement showed plateau in five lesions and washout in one lesion. MRI BI-RADS categories were distributed as follows: one BI-RADS II, one BI-RADS III, three BI-RADS IV and two BI-RADS V. CONCLUSIONS: MRI features of primary breast lymphomas in this study cohort suggest that the occurrence of a PBNHL should be considered in the presence of large enhancing lesions of the breast, especially if associated with skin thickening. MRI may also have an important role in the assessment of response to therapy and diagnosis of recurrence. PMID- 19562269 TI - Molecular characterization of the niaD and pyrG genes from Penicillium camemberti, and their use as transformation markers. AB - Genetic manipulation of the filamentous fungus Penicillium camemberti has been limited by a lack of suitable genetics tools for this fungus. In particular, there is no available homologous transformation system. In this study, the nitrate reductase (niaD) and orotidine-5'-monophosphate decarboxylase (pyrG) genes from Penicillium camemberti were characterized, and their suitability as metabolic molecular markers for transformation was evaluated. The genes were amplified using PCR-related techniques, and sequenced. The niaD gene is flanked by the nitrite reductase (niiA) gene in a divergent arrangement, being part of the putative nitrate assimilation cluster in P. camemberti. pyrG presents several polymorphisms compared with a previously sequenced pyrG gene from another P. camemberti strain, but almost all are silent mutations. Southern blot assays indicate that one copy of each gene is present in P. camemberti. Northern blot assays showed that the pyrG gene is expressed in minimal and rich media, and the niaD gene is expressed in nitrate, but not in reduced nitrogen sources. The functionality of the two genes as transformation markers was established by transforming A. nidulans pyrG- and niaD-deficient strains. Higher transformation efficiencies were obtained with a pyrG-containing plasmid. This is the first study yielding a molecular and functional characterization of P. camemberti genes that would be useful as molecular markers for transformation, opening the way for the future development of a non-antibiotic genetic transformation system for this fungus. PMID- 19562268 TI - Computation of brain metabolite ratios in single-voxel proton MR spectroscopy: comparison between semiautomatic and automatic software. AB - PURPOSE: Metabolite ratios are the measurements most commonly utilised for clinical applications of brain proton magnetic resonance spectroscopy ((1)H-MRS) [1]. We evaluated the agreement between the metabolite ratios calculated with semiautomatic and automatic software. MATERIALS AND METHODS: Two single-voxel spectra (3.375 ml) localised in the frontal grey matter (GM) and peritrigonal white matter (WM) were obtained in 20 healthy subjects by using a point-resolved proton spectroscopy sequence (PRESS, TE=144 ms). The spectra were processed using the semiautomatic software J-Magnetic Resonance User Interface (JMRUI) and the automatic software SpectroView. Agreement of the N-acetyl-aspartate (NAA)/creatine (Cr), NAA/choline (Cho) and Cho/Cr ratios calculated with the two methods was assessed by estimating the 95% limits of agreement (LAs) of the differences of the values obtained with the two software packages. RESULTS: Mean values and standard deviations of NAA/Cr, Cho/Cr and NAA/Cho (semiautomatic//automatic software) were 1.99+/-0.53//1.73+/-0.36, 1.13+/ 0.40//1.04+/-0.33, 1.85+/-0.62//1.89+/-0.69 for the GM and 2.24+/-0.41//2.37+/ 0.27, 0.96+/-0.17//1.13+/-0.15, 2.37+/-0.43//2.11+/-0.23 for the WM. The 95% LAs were wider for GM spectra and ranged between -0.51, 0.17 for Cho/Cr in the WM and -1.54, 1.47 for NAA/Cho in the GM. CONCLUSIONS: The difference between brain metabolite ratios calculated with the two software packages is not negligible and reflects spectral quality. PMID- 19562270 TI - Overexpression of human osteopontin increases cell proliferation and migration in human embryo kidney-293 cells. AB - Malignant tumors are characterized by dysregulated cell growth and the metastasis of secondary tumors. Numerous studies have documented that osteopontin (OPN) plays a key role in regulating tumor progression and metastasis. Here, we show that the overexpression of OPN in human embryo kidney-293 cells significantly increases both the level of cell proliferation, by provoking the G(1)/S transition, and the level of cell migration in vitro. These findings suggest that augmented OPN contributes to cell growth and motility. Inhibiting OPN or the pathway it stimulates may therefore represent a novel approach for the treatment of primary tumors and associated metastases. PMID- 19562271 TI - The transcriptional regulation of podocin (NPHS2) by Lmx1b and a promoter single nucleotide polymorphism. AB - Podocin (NPHS2) is a component of the glomerular slit membrane with major regulatory functions in the renal permeability of proteins. A loss of podocin and a decrease in its resynthesis can influence the outcome of renal diseases with nephrotic syndrome, such as minimal change glomerulonephritis, focal segmental glomerulosclerosis (FSGS) and membranous nephropathy. The transcriptional regulation of podocin may play a major role in these processes. We defined the transcriptional regulation of the human podocin gene and the influence of single nucleotide polymorphisms (SNPs) within its promoter region in the podocytes using reporter gene constructs and gel shift analysis. In addition, we took genomic DNA from healthy Caucasian blood donors and from biopsies of kidneys with defined renal diseases and screened it for podocin promoter SNPs. Our data shows that the transcription of podocin is mainly regulated by the transcription factor Lmx1b, which binds to a FLAT-F element and displays enhancer function. With the SNP variant -116T, there was a significant reduction in luciferase activity, and nuclear protein binding was observed, while the SNP -670C/T did not display functionality. The allelic distribution of -116C/T in patients with kidney diseases leading to nephrotic syndrome was not significantly different from that in the control group. Our data indicates that among other factors, podocin is specifically regulated by the transcription factor Lmx1b and by the functional polymorphism -116C/T. However, there is no association between -116C/T and susceptibility to minimal change glomerulonephritis, focal segmental glomerulosclerosis or membranous nephropathy. PMID- 19562272 TI - Copper and zinc in the serum, urine, and hair of patients with Wilson's disease treated with penicillamine and zinc. AB - The purpose of this study was to determine the different levels of copper and zinc in the serum, urine, and scalp hair of patients with Wilson's disease receiving different, currently accepted methods of treatment to reduce the copper load (penicillamine-group 1, n = 8; zinc-group 2, n = 8; penicillamine+zinc-group 3, n = 8). Blood, urine, and hair samples were collected from the patients. All three treatments resulted in a significant decrease of the serum copper levels. Significantly increased levels of zinc in the serum were detected in the patients in groups 2 and 3 (19.1 and 18.8 micromol/l, respectively; p < 0.05). Copper excretion in the urine significantly increased during its administration to groups 1 and 3 (11.5 and 7.94 micromol/24 h respectively; p < 0.001) due to the effect of penicillamine. The administration of zinc as monotherapy (group 2) or in combination with penicillamine (group 3) led to an increase of its excretion (25.3 and 22.4 micromol/24 h, respectively; p < 0.01). Only an insignificant rise of the copper content in the hair was found in all three groups of patients. The content of zinc in the hair did not differ significantly in any of the groups in comparison with the control group. PMID- 19562273 TI - Staphylococcus aureus colonization of anterior nares of school going children. AB - OBJECTIVE: To determine the colonization rates of S. aureus in anterior nares of school going children, evaluate the antimicrobial resistance of such isolates against various antibiotics. METHODS: Nasal swabs taken for S. aureus in 392 healthy school going children aged between 5 and 15 yr belonging to three schools surrounding Kamineni Institute of Medical Sciences, Narketpally, Andhra Pradesh, India. Swabs were inoculated in to Mannitol Salt Agar (MSA) and incubation at 37 degrees C for 24hr, Staphylococcus aureus isolates were identified by standard microbiological methods such as Gram's stain, catalase and coagulase. MICs were determined by Agar dilution technique against Vancomycin, Ciprofloxacin, Sparfloxacin, Sparfloxacin beta Cyclodextrin. Antimicrobial resistance patterns of all the isolates against Oxacillin (1 microg) Penicillin (10 units), Ampicillin (10 microg), Co-trimoxazole (23.75 microg), Erythromycin (15 microg), Tetracycline (30 microg) and Gentamicin (10 microg) were tested using Kirby-Baeur disk diffusion method in accordance with CLSI standards. RESULTS: Of the 392 samples screened 63(16%) showed the growth of Staphylococcus aureus. 12(19%) isolates were found to be MRSA. Antimicrobial susceptibility testing using Agar dilution method against Vancomycin, ciprofloxacin, Sparfloxacin, and Sparfloxacin beta Cyclodextrin revealed MICs in the range of 0.52 microg/ml, 0.51 microg/ml, 0.5 microg/ml and <0.0312-0.250 microg/ml respectively. Disk diffusion method showed that all the isolates were resistant to Penicillin and Ampicillin. A resistance of 14.3%, 25% and 22.2% was observed against Co-trimoxazole, Erythromycin and Tetracycline respectively. Gentamiicin was the only antibiotic against which most of the isolates were sensitive. CONCLUSION: Our results suggests that healthy school going children under 16 yr of age are potential carriers of S. aureus and in particular MRSA and multi-drug resistant strains. PMID- 19562274 TI - Long-acting risperidone: a review of its role in the treatment of bipolar disorder. AB - Bipolar disorder is a multidimensional illness typified by fluctuating periods of depression and mania, cognitive dysfunction, abnormal circadian rhythms, and multiple comorbid psychiatric and general medical conditions. Indefinite pharmacological treatment is often required, yet the modest effects of available treatments and frequent difficulties with tolerability and adherence present complex challenges to patients. Long-acting injectable medications offer a therapeutic alternative to oral mood stabilizers and may help facilitate long term treatment adherence. This article will provide a succinct review of the latest data on the use of long-acting injectable risperidone (LAR) during the maintenance-phase treatment of bipolar disorder. The specific role of LAR in comparison to other atypical antipsychotics, and the limitations of available studies will be discussed from the perspectives of efficacy, tolerability, and sequential positioning in treatment guidelines. PMID- 19562275 TI - Montelukast in guidelines and beyond. AB - In all asthma guidelines, preventive anti-inflammatory treatment is essential in all patients with persistent asthma. Inhaled corticosteroids are the mainstay of treatment in the control of asthma, but other treatments may be used as a monotherapy in patients with mild asthma or as an add-on treatment in those with moderate-to-severe asthma. Leukotriene modifiers are the only validated preventive treatment for all age groups. This review discusses the place of montelukast, a leukotriene receptor antagonist, using guidelines and consensus reports on asthma and rhinitis: the US National Asthma Education and Prevention Program (NAEPP); the British Guideline on the Management of Asthma; the Global Initiative on Asthma (GINA); and Allergic Rhinitis and its Impact on Asthma (ARIA). This review includes new studies that have not yet been considered in guidelines. PMID- 19562277 TI - ABO-incompatible kidney transplantation. PMID- 19562278 TI - [Spirituality, religion and culture on the intensive care unit--how are these compatible?]. PMID- 19562280 TI - Intrathoracal displacement of the humeral head after fracture. PMID- 19562279 TI - Parathyroid hormone-dependent hypercalcemia. AB - The past fifteen years have resulted in great progress in our understanding of the pathogenesis and pathophysiology of hypercalcemic disorders occurring either sporadically or in a familial setting. This paper briefly reviews the clinically most important new knowledge on sporadic and hereditary forms of parathyroid hormone-dependent hypercalcemic disorders, with special emphasis on familial syndromes such as multiple endocrine neoplasia type 1 and type 2A, hyperparathyroidism-jaw tumor syndrome, familial isolated hyperparathyroidism, familial hypocalciuric hypercalcemia and neonatal severe primary hyperparathyroidism. In addition, the authors briefly present the most important clinical characteristics of 141 patients with parathyroid hormone-dependent hypercalcemia, including index patients of 18 families with hereditary disorders, diagnosed in a Hungarian endocrine center between 1997 and 2007. PMID- 19562282 TI - Medical teachers' attitudes towards science and motivational orientation for medical research. AB - BACKGROUND: Research is an important motivating factor for pursuing a career in academic medicine, but the relation between motivation and other factors involved in scientific research are not clear. PURPOSE: To explore the motivational orientation for doing research and its relation with attitudes towards science and publication practice among members of faculty at a medical school. METHODS: We used a Science Attitude Survey and the Work Preference Inventory (intrinsic and extrinsic motivational orientation using 4 Likert-type scales of motivation, possible range 1-5) to survey two groups of teachers at the Zagreb University School of Medicine (n = 327, 66% response rate): professors, elected to tenure track positions (n = 150), and instructor/research fellows working on or just completing their thesis (n = 177). RESULTS: Overall, teachers scored highest on the Enjoyment subscale of intrinsic motivational orientation (mean score +/- standard deviation 4.3 +/- 0.42 for professors vs 4.1 +/- 0.42 for instructors/research fellows, P = 0.001, t-test). Professors also scored higher than instructors/research fellows on the Challenge subscale of intrinsic motivational orientation (3.8 +/- 0.55 vs. 3.5 +/- 0.64, P < 0.001, t-test), whereas instructors/research fellows scored higher on the Compensation subscale of extrinsic motivational orientation (3.5 +/- 0.74 vs. 3.1 +/- 0.71, P < 0.001, t-test). Multiple linear regression analysis showed that the number of publications was positively associated with scores on the Science Attitude Survey and the Challenge subscale of intrinsic motivation, and negatively associated with scores on the Compensation subscale of extrinsic motivation. CONCLUSIONS: Members of the medical faculty differ in motivational orientation for research depending on their academic status, and their motivation is associated more with requirements for academic advancement than with research. These findings have important implications for developing strategies for enhancing academic research production. PMID- 19562281 TI - Vienna experience of ABO-incompatible living-donor kidney transplantation. AB - ABO-incompatible kidney transplantation is a promising strategy for enlargement of living-donor pools. In recent years, recipient desensitization by blood group antigen-specific immunoadsorption, together with rituximab and intravenous immunoglobulin, has allowed excellent graft performance after ABO-incompatible transplantation. Adopting this protocol, originally described by Tyden and coworkers, we performed four living-donor renal transplants across the ABO barrier (A1-->0, A1-->B, B-->A1, A2-->0) between July 2007 and August 2008. Recipients were aged 25-66 years, donors 49-69 years. A protocol of on-demand immunoadsorption was followed, based on serial post-transplant antibody monitoring. Substantial and sustained decrease of blood group antibody levels was achieved in all four recipients, therefore post-transplant immunoadsorption was not needed. Graft and patient survival after 4-18 months' follow-up was 100%. Current serum creatinine was 1.3-2.0 mg/dl. Two grafts showed C4d deposits in peritubular capillaries in the complete absence of typical morphological features of antibody-mediated rejection. One recipient experienced early graft dysfunction, diagnosed as Banff borderline lesion, which responded well to steroid pulse therapy. The same recipient developed de novo interstitial fibrosis/tubular atrophy and arteriolar hyalinosis, presumably the result of suboptimal control of blood pressure and/or calcineurin inhibitor therapy. Two of the four recipients developed lymphoceles necessitating surgical revision. Apart from urinary tract infection in three patients and subclinical CMV in one, no major infectious complications were reported. Notably, two stable recipients developed polyoma BK viremia without clinical or morphological manifestations of polyomavirus-associated nephropathy. The results obtained in our small series support the earlier reported high efficiency of desensitization based on antigen specific immunoadsorption. Nevertheless, the lack of long-term data will necessitate continuous and prudent consideration of the benefits and risks of this strategy. PMID- 19562284 TI - Cholesterol metabolism in active Crohn's disease. AB - Hypocholesterolemia has been investigated as a typical feature of critical illness and is connected with poor prognosis. Crohn's disease is an inflammatory process and is associated with several metabolic disturbances. In recent decades clinical studies have established a link between lipid metabolism and systemic inflammation. In our study we examined the serum profile of cholesterol (total cholesterol, LDL- and HDL-cholesterol) and changes in the cholesterol absorption/synthesis process by determination of plasma non-cholesterol sterol (squalene, lathosterol, campesterol, sitosterol) concentrations. Serum concentrations of total cholesterol, LDL- and HDL-cholesterol and non-cholesterol sterols were evaluated in 24 patients with active Crohn's disease during a period of 28 days. We detected lower serum levels of total cholesterol (P < 0.001), LDL- and HDL-cholesterol (P < 0.05) in the patients with active Crohn's disease than in the control group. In addition, the patients had significantly lower plasma levels of lathosterol (P < 0.001) and higher concentrations of squalene, although without significant differences. A significant decrease of campesterol plasma levels (P < 0.001) was detected, but lower plasma concentrations of sitosterol were without statistical significance. The active phase of Crohn's disease is characterized by altered metabolism of lipids, mainly of cholesterol. Our results show abnormalities in plasma concentrations of non-cholesterol sterols and provide evidence that the process of cholesterol synthesis and absorption is altered in active Crohn's disease. PMID- 19562283 TI - Subcutaneous adipose tissue topography and metabolic disturbances in polycystic ovary syndrome. AB - OBJECTIVE: Polycystic ovary syndrome (PCOS) is the most common endocrine disorder affecting women of reproductive age. Central obesity plays a major role in the pathophysiology of PCOS. However, there is little information on the impact of subcutaneous adipose tissue (SAT) on metabolic disturbances in PCOS. The aim of this study was to investigate whether SAT topography influences insulin resistance, impaired glucose tolerance and metabolic parameters in women with PCOS. METHODS: 36 women aged 16-41 years with PCOS and 87 healthy women aged 20 34 years were examined using lipometry, metabolic and hormonal measurements, oral glucose tolerance tests, hirsutism scores, and questionnaires. The homeostasis model assessment (HOMA) index was used for determination of insulin resistance. RESULTS: SAT measurement points on the trunk showed significant positive correlation with the HOMA index. A negative correlation between calf SAT and the HOMA index was seen. Multiple regression analysis detected a positive association between the HOMA index and lower-abdomen SAT and upper-back SAT, whereas hip SAT showed a negative association with the HOMA index. In overweight/obese patients with PCOS, lower-abdomen and upper-back SAT showed significant positive correlations with insulin resistance. There was no correlation of SAT topography with insulin resistance in lean women with PCOS. Compared with PCOS women with normal glucose tolerance, patients with glucose intolerance had significantly increased trunk obesity and decreased leg fat. Increased SAT layers on the trunk were related to an unfavorable serum lipid profile, whereas increased leg fat correlated positively with HDL cholesterol. CONCLUSIONS: Increased SAT layers on the trunk are associated with insulin resistance, impaired glucose tolerance and an unfavorable lipid profile in women suffering from PCOS. Increased thickness of leg SAT emerges as being protective against metabolic disturbances in PCOS. PMID- 19562285 TI - Prevalence and risk factors for voice problems in priests. AB - PURPOSE: Voice problems are common in occupations with a large voice loading. Clergy are usually classified as professional voice users. The aim of our study was to investigate the prevalence of voice problems in Catholic priests and to identify some of the risk factors for their voice disorders. METHODS: Questionnaires were sent to 600 randomly chosen Catholic parish priests and vicars in Slovenia to collect information on the prevalence and causes of voice disorders, vocal load, vocal habits, and certain diseases influencing voice quality. Data were analyzed for a total of 340 priests and in subgroups of priests with frequent voice disorders, priests without voice disorders, and priests over the age of 64. RESULTS: Overall, 85.6% of the priests reported having voice problems during their career, 15.9 % of them experiencing frequent voice problems. Respiratory-tract infection was the most common cause of the voice problems. The following significant risk factors for frequent voice problems were identified: frequent throat clearing, vocal load during spare time, voice disorders during training and not receiving instruction on correct vocal technique. The priests without voice problems had experienced significantly fewer voice problems during their training and had less frequent throat clearing, less frequent asthma and fewer allergies than the other priests. Being over the age of 64 and teaching more than 10 hours each week were not identified as risk factors for voice disorders. CONCLUSIONS: The prevalence of voice disorders in Catholic priests in Slovenia is high. Respiratory-tract infection is the most common cause of their voice problems. Allergies, frequent throat clearing, not receiving instruction on correct vocal use and vocal hygiene, and a history of voice problems during training were stated as risk factors for voice disorders. In order to decrease the prevalence of voice disorders in priests and in other occupations with a large vocal load, we suggest that an ENT examination before the start of training and the inclusion of information on correct use of the voice and on diseases influencing voice quality should be included as a standard part of the educational process. PMID- 19562286 TI - Mycobacterium microti--pulmonary tuberculosis in an immunocompetent patient. AB - Human tuberculosis is caused by members of the Mycobacterium tuberculosis complex, which includes M. tuberculosis, M. bovis, M. africanum and M. bovis BCG. However there are increasing reports of rarely occurring genetic variants such as M. canettii, M. microti and M. pinipedii. The natural reservoir, mode of transmission and potential modification of host interaction of these species is not yet fully elucidated. We report a rare case of extensive cavitary smear positive tuberculosis of the left lung caused by M. microti in an immunocompetent tuberculin-negative 68-year-old man. Transmission by a raccoon dog or raccoon as a novel M. microti reservoir was suspected. Spoligotyping of the isolate revealed the llama subtype. The strain exhibited no detectable drug resistance. Response to standard tuberculosis treatment, initially comprising isoniacid, rifampicin, pyrazinamide and ethambutol, was excellent. Delayed growth on solid media, specific phenotypic features and contact with animals should raise suspicion for this rare mycobacterial infection. PMID- 19562288 TI - COPD is not COPD is not allergy. AB - Chronic obstructive pulmonary disease (COPD) is still a poorly understood disease. Its pathogenesis is excitingly complex and has systemic consequences caused not only by increased production of certain cytokines but also by neurohumoral activation, chronic bacterial infection, muscle wasting and cachexia. Asthma and COPD have many overlapping clinical features so it should not be surprising that in the pathogenesis of COPD mediators such as leukotrienes, complement activation, atopic or even autoimmune processes are possibly involved. The pathogenesis of cardiovascular system involvement in COPD is also multifaceted and includes chronic heart hypoxia, damage by smoking and pulmonary hypertension; it must also be viewed as a consequence of systemic inflammation and neurohormonal activation. COPD is among the leading causes of morbidity and mortality worldwide and therefore it should be studied intensively beyond the lung itself. Treatments directed at neurohumoral activation in COPD have not been fully addressed; this aspect of COPD should be better understood, as it may direct novel therapeutic approaches. PMID- 19562289 TI - Neurohormonal activation and inflammation in chronic cardiopulmonary disease: a brief systematic review. AB - Chronic cardiopulmonary disease typically induces and maintains (over)activation of several phylogenetically old adaptational and defensive mechanisms. Activation was usually needed for a limited period during acute danger or injury. In chronic disease conditions, however, those mechanisms are kept activated for longer periods. Eventually, irreversible damage is done and this contributes to impaired function and worse prognosis in a variety of chronic disease. Landmark trials in chronic heart failure have provided robust evidence for prognostic benefit for neurohormonal antagonists. Retrospective and epidemiological data for their beneficial effect in chronic obstructive pulmonary disease begin to accumulate and new fields (e.g. cancer and stroke) could be pending in the future. PMID- 19562290 TI - Distribution of self-rated health and association with clinical parameters in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Data on self-rated health (SRH) in patients with chronic obstructive pulmonary disease (COPD) are very limited; we therefore initiated this study to investigate the distribution of SRH and association with established parameters of disease severity. PATIENTS AND METHODS: We included 135 clinically stable patients with COPD (64 +/- 8 years, 71% men, GOLD stage: II -59; III -55; IV -21) and 25 healthy control persons. SRH was evaluated using the 5-grade Likert scale (1-very poor to 5-very good). RESULTS: Patients with COPD had poorer SRH when compared with controls (3.0 +/- 0.7 vs. 3.8 +/- 0.6, P < 0.001). SRH decreased over GOLD stage (P = 0.016) and 27 (20%) patients reported poor or very poor SRH. In univariate analysis, GOLD stage (P = 0.022), Center for Epidemiologic Studies Depression (CES-D) score (P = 0.001), BODE index score (P < 0.001), score on the modified Medical Research Council (MMRC) dyspnea scale (P < 0.001) and 6-minute walk test (6MWT) distance (P < 0.001) determined poor or very poor SRH. In a multivariate model which included BODE index score, a CES-D score > or = 16 (P = 0.013) and BODE index score (P = 0.012) determined poor or very poor SRH. In the model with individual components of the BODE index, a CES-D score > or = 16 (P = 0.012), MMRC score of 3 or 4 (P = 0.019) and 6MWT distance < or = 249 m (P = 0.019) determined poor or very poor SRH. CONCLUSION: In patients with COPD, SRH is worse than in healthy control persons and deteriorates over GOLD stage. Perception of health as poor or very poor is associated with psychological components (CES-D score) and disease severity (BODE index score, 6MWT distance and MMRC dyspnea score). PMID- 19562291 TI - Elevated TNFalpha production in whole blood in patients with severe COPD: the potential link to disease severity. AB - OBJECTIVE: The relationship between tumour necrosis factor-alpha (TNFalpha), severity of pulmonary disease and nutritional depletion in chronic obstructive pulmonary disease (COPD) remains unclear. We aimed to clarify the role of lipopolysaccharide (LPS) as a potential stimulus of cytokine production and the role of these cytokines in the alteration of body composition in patients with different degrees of COPD. PATIENTS AND METHODS: We studied 29 weight-stable out patients with different severites of COPD who had no evidence of recent infection or significant co-morbidity. Baseline serum TNFalpha levels and TNFalpha response to LPS in whole blood were measured in patients and 20 aged matched controls. RESULTS: Serum TNFalpha was significantly elevated in patients versus controls (2.1 +/- 0.3 vs. 1.1 +/- 0.1 pg/ml, mean +/- SEM, P = 0.007). In patients with COPD, we found a significant correlation between serum TNFalpha levels and disease severity, assessed as FEV(1) %predicted (r = 0.49, P = 0.02). Response to lipopolysaccharide did not differ significantly between patients and controls. However, within the patient group those with more severe disease (FEV(1) < or = 30% predicted, n = 12) had an enhanced response compared to patients with mild-to moderate disease (all P < 0.05 for LPS > 1 ng/ml). Spontaneous TNFalpha production was 5.0 times higher in patients with severe COPD compared to mild-to moderate COPD (P = 0.02). There was no relation between body composition and serum TNFalpha or TNFalpha response to LPS. CONCLUSION: Increasing airflow obstruction and hypercapnia are associated with an enhanced TNFalpha response in COPD. PMID- 19562293 TI - Preoperative total serum cholesterol as a prognostic factor for survival in patients with resectable non-small-cell lung cancer. AB - BACKGROUND: Coexisting hypocholesterolemia in cancer has been known for a very long time but its relationship to cancer is still controversial. Hypocholesterolemia has been reported in patients with lung cancer, although its association with survival has not been explored. OBJECTIVES: The purpose of the study was to determine whether preoperative total serum cholesterol is a prognostic factor for survival after lung cancer resection. METHODS: The retrospective study comprised 198 patients (162 men, 36 women) operated upon for resectable non-small-cell lung cancer (clinical stages I-IIIB) between January 1992 and April 1994. Total serum cholesterol concentration was determined preoperatively in each patient. The effects of sex, age, stage, histological type and preoperative total serum cholesterol concentration on survival were tested in univariate and multivariate analysis. RESULTS: Preoperative total serum cholesterol was a significant prognostic factor in both univariate and multivariate analysis. The median value for total serum cholesterol was 5.3 mmol/l and patients below that cut-off had significantly shorter overall survival times than patients in the high cholesterol group (5-year survival 41% vs. 56%, P < 0.05). In a multivariate Cox proportional-hazard regression model, only stage and preoperative total serum cholesterol were found to be of significance for survival (relative risk 0.84 for each mmol/l increase in concentration, CI 0.71 1.00, P < 0.05). CONCLUSIONS: Our results suggest that preoperative total serum cholesterol may be an important prognostic factor for overall survival after lung cancer resection. It may prove to be a valuable tool in the follow-up of patients with lung cancer and in detection of high-risk cases. PMID- 19562292 TI - The burden of chronic obstructive pulmonary disease in patients hospitalized with heart failure. AB - OBJECTIVES: Like chronic heart failure, chronic obstructive pulmonary disease (COPD) is an enormous public health problem in industrialized countries. Our aim was to determine the prevalence and clinical impact of COPD among patients hospitalized for heart failure in a community hospital serving a population of 125,000 people. METHODS: Between 2001 and 2003 a total of 638 patients (73 +/- 10 years, 48% men, 74% NYHA class III) were identified with a discharge diagnosis of heart failure. Medical charts were reviewed and vital status was obtained from a Central Population Registry. RESULTS: COPD was diagnosed in 106 (17%) patients whose age was similar to those without COPD (73 +/- 9 vs. 73 +/- 11 years, P = 0.35). Patients with COPD were more often males (65% vs. 45%, P < 0.001). There were no differences in arterial hypertension, atrial fibrillation, diabetes mellitus and most laboratory markers except hemoglobin (141 +/- 20 vs. 132 +/- 20 g/l, P < 0.001) and uric acid (453 +/- 136 vs. 414 +/- 139 mmol/l, P = 0.013). At discharge, patients with COPD were less likely to receive beta-blockers (12% vs. 28%, odds ratio 0.35, 95% CI0.19-0.64). During follow-up, patients with COPD had higher mortality (73% vs. 60%, P = 0.016, hazard ratio 1.48, 95% CI 1.15-1.90). Kaplan-Meier (log-rank test, P = 0.002) and Cox proportional hazard analysis, adjusted for age, sex, hemoglobin, uric acid, and treatment with beta-blockers and furosemide (hazard ratio 1.38, 95% CI1.04-1.83, P = 0.024) demonstrated the prognostic importance of COPD. CONCLUSIONS: COPD is frequent among hospitalized patients with heart failure. Beta-blockers are largely underused, which is probably a major reason for the higher mortality observed in patients with concomitant chronic heart failure and COPD. PMID- 19562294 TI - Venous thromboembolism prophylaxis in hospitalized patients with pneumonia: a prospective survey. AB - BACKGROUND: Guidelines for prevention of venous thromboembolism recognize pneumonia and changes in respiratory status as risk factors. There is little information on the preventive use of low-molecular-weight heparin (LMWH) in hospitalized patients with pneumonia. METHODS: We prospectively screened 1067 admissions to our hospital for preventive use of LMWH according to the American College of Chest Physicians (ACCP) guidelines. The analysis included 168 patients with pneumonia (age 74 +/- 16 years, 56% men). The primary and secondary outcomes were treatment with LMWH in eligible patients and LMWH use according to guidelines (daily dose, duration of treatment). RESULTS: LMWH use was indicated in 126 (75%) patients and 119 (94%) were actually treated. In 41% of patients treatment was according to the ACCP guidelines. The dose and duration of LMWH treatment were appropriate in 61% and 66% of patients, respectively. Non-use of LMWHs was not associated with clinical and demographic characteristics. Adverse effects included bleeding (N = 7) and thrombocytopenia (N = 2) but were not associated with fatality. Prolonged treatment with LMWH was associated with adverse effects (P < 0.05). CONCLUSIONS: Implementation of LMWH prophylaxis for venous thromboembolism in hospitalized patients with pneumonia reached 94%. Adherence to ACCP guidelines was complete in 41% of patients. Prolonged treatment with LMWH was associated with non-fatal adverse effects, which calls for timely withdrawal of LMWH once no longer indicated. PMID- 19562295 TI - Ventricular arrhythmic disturbances and autonomic modulation after beating-heart revascularization in patients with pulmonary normotension. AB - BACKGROUND: De-novo ventricular arrhythmias are potentially life-threatening complications after beating-heart revascularization (off-pump CABG). Whether pulmonary hypertension can influence initiation of ventricular arrhythmias through increased sympathetic activity is controversial. In order to determine the influence of pulmonary hypertension on its relative contribution to ventricular arrhythmia, we first had to define the role of cardiac autonomic modulation in patients with pulmonary normotension. We aimed to observe how parameters of linear and nonlinear heart rate variability are changed pre- and postoperatively in patients with pulmonary normotension undergoing off-pump CABG. METHODS: Fifteen-minute ECG recordings were collected before and after off-pump CABG in 54 patients with multivessel coronary artery disease and pulmonary normotension to determine linear (TP, HF, LF, LF:HF ratio) and nonlinear detrended fluctuation analysis (alpha1, alpha2) and fractal dimension (average, high and low) parameters of heart rate variability. Arrhythmia was monitored preoperatively in 24-hour Holter recordings and postoperatively by continuous monitoring and clinical assessment. RESULTS: Deterioration from simple (Lown I II) to complex (Lown III-V) ventricular arrhythmia was observed in 19 patients, and improvement from complex to simple arrhythmia in five patients (P = 0.022). Patients with postoperative deterioration of ventricular arrhythmia had preoperatively significantly lower values of TP, HF and LF (P = 0.024-0.043) and postoperatively significantly higher values on the low fractal dimension index (P = 0.031) than patients with postoperative improvement of arrhythmia. CONCLUSION: Patients experiencing postoperative deterioration of ventricular arrhythmia already have impaired autonomic regulation before surgery. Higher postoperative values on the low fractal dimension index indicate that sympathetic predominance with or without concomitant vagal withdrawal is the underlying neurogenic mechanism contributing to ventricular arrhythmia. PMID- 19562296 TI - Heart rate-corrected QT interval and QT dispersion in patients with pulmonary hypertension. AB - OBJECTIVE: Heart rate-corrected QT interval (QTc) and QTc dispersion (QTcd) are increased and associated with ventricular arrhythmia and an increase in sudden death in a variety of diseases. This study aimed to examine QTc and QTcd in pulmonary hypertension and assess their relationship with pulmonary arterial pressure. PATIENTS AND METHODS: A total of 201 patients who had undergone right heart catheterization for a preliminary diagnosis of pulmonary hypertension between December 2003 and July 2008 were included in the study. Resting 12-lead electrocardiogram was recorded. QT interval was measured manually and corrected using Bazett's formula. Patients were divided into groups with mild-to-moderate and severe pulmonary hypertension and a control group according to mean pulmonary arterial pressure. RESULTS: In all observed cases, mean QTc was higher in severe pulmonary hypertension than in controls (428.6 +/- 32.8 ms vs. 411.1 +/- 28.4 ms, P = 0.018) and QTcd was higher in mild-to-moderate (60.1 +/- 17.2 ms) and severe pulmonary hypertension (63.9 +/- 20.5 ms) than in controls (47.3 +/- 10.6 ms) (P = 0.031; P = 0.004). In men, there was no significant difference in mean QTc and QTcd. In women, mean QTc was higher in severe pulmonary hypertension than in controls (436.1 +/- 39.4 ms vs. 407.6 +/- 24.8 ms, P = 0.037) and QTcd was higher in severe pulmonary hypertension (68.5 +/- 20.9 ms) than in both the controls (45.1 +/- 12.6 ms) and patients with mild-to-moderate pulmonary hypertension (58.6 +/- 14.7 ms) (P = 0.002; P = 0.003). In addition, in women with pulmonary hypertension, mean QTc and QTcd were positively correlated to mean pulmonary arterial pressure (r = 0.207, P = 0.03; r = 0.236, P = 0.012). CONCLUSIONS: In women with pulmonary hypertension, mean QTc and QTcd are positively correlated to mean pulmonary arterial pressure and are significantly increased in patients with severe pulmonary hypertension. PMID- 19562297 TI - Sonographic evaluation of unexplained pleural exudate: a prospective case series. AB - BACKGROUND: Thoracic ultrasound may be helpful in differentiating between malignant and tuberculosis-associated pleural exudate. This study aimed to evaluate its utility in patients with unexplained pleural exudate. METHODS: Consecutive patients were screened and pleural effusion was found in 278 patients. Pleural exudate was present in 106 patients and remained undiagnosed after biochemical and cytological evaluation in 40 patients (median age 58 years, 67% men) who then underwent detailed thoracic ultrasound for the presence of complex (septated or fibrous) or anechoic patterns. Pleural needle biopsy or thoracoscopy with histological evaluation were used for definitive diagnosis. RESULTS: History, clinical characteristics and routine procedures including cytology were not helpful in differential diagnosis. Pleural specimens for histological evaluation were obtained from all 40 patients and confirmed tuberculosis in 12 patients, cancer in nine and nonspecific pleuritis in 19. Sonographic finding of a complex septal pattern was present only in patients with tuberculosis (positive predictive value 100%); anechoic appearance was suggestive of nonspecific pleuritis (positive predictive value 65%). CONCLUSIONS: Thoracic ultrasound is a useful bedside method for differentiation of the etiology of pleural exudate. When a complex septal pattern is found, pleural needle biopsy should be the next diagnostic procedure, whereas with less complex pleural sonography findings other methods should be pursued. PMID- 19562298 TI - C5a-induced in vitro basophil activation in patients with chronic urticaria: a pilot study. AB - BACKGROUND: Complement component 5a (C5a) might be involved in the formation of wheals in patients with chronic urticaria (CU). We sought to compare the in vitro responsiveness of basophils to C5a in patients with CU and in a control group. METHODS: Basophil surface expression of activation marker CD63 induced by C5a, anti-FcepsilonRI mAb or anti-IgE pAb was measured using flow cytometry in 17 patients with CU and in 10 healthy controls. RESULTS: Patients with CU showed significantly greater basophil CD63 surface expression induced by C5a (median [interquartile range]; 16.4% [13-25.1]; P = 0.011) than the group of healthy controls (10.7% [7.2-16.8]). In contrast, basophil CD63 response to anti-IgE and anti-FcepsilonRI was lower in the CU group (12.3% [6-36.3]; 25.9% [12.5-60.5]) than in the control group (51.7% [6.7-84.3]; 62.1% [9.7-89.2]), although not statistically significant. CONCLUSION: Results of this pilot study suggest that patients with CU might have an enhanced basophil response to stimulation with C5a, indicating that further studies in CU basophil responsiveness are needed. PMID- 19562299 TI - Diagnostic value of the basophil activation test in evaluating Hymenoptera venom sensitization. AB - BACKGROUND: Diagnosis of allergy to Hymenoptera venom is usually confirmed with skin testing and measurement of specific serum IgE antibody, tests which are sometimes inconclusive. In these cases, additional in vitro tests are necessary. The aim of this study was to show the applicability of the basophil activation test in detecting sensitization to Hymenoptera venom and to compare the test sensitivity and clinical positive-predictive value with skin prick tests and measurement of allergen-specific serum IgE. METHODS: This prospective study was conducted between June 2004 and December 2007 and included a large group of 204 patients. All patients had a history of at least one systemic allergic reaction of Muller grades II-IV after a Hymenoptera sting. We compared results of the basophil activation test, specific serum IgE and skin prick tests with patients' clinical history and data on culprit insects. RESULTS: The overall clinical sensitivities of the basophil activation test, specific serum IgE and skin prick tests were 90%, 76% and 64%, respectively; the clinical positive-predictive values of the three tests were 79%, 73% and 78% for bee venom, 86%, 59% and 43% for wasp venom; and 84%, 77% and 22% for both venoms. CONCLUSIONS: Our results revealed a higher clinical sensitivity and comparable or better clinical positive predictive value of basophil activation tests than skin prick tests and allergen specific serum IgE in the detection of allergy to Hymenoptera venom. PMID- 19562300 TI - Carbohydrate epitopes as a cause of cross-reactivity in patients allergic to Hymenoptera venom. AB - BACKGROUND: Among patients with allergy to insect stings, double positivity in tests for IgE antibodies specific to honey bee and wasp venoms is a frequent diagnostic problem. True double sensitization and possible cross-reactivity of venom hyaluronidases and with carbohydrate determinants must be considered in such patients. We studied the frequency of sensitization to carbohydrate determinants and the role of these in double positivity in tests for specific IgE antibodies. MATERIALS AND METHODS: A group of 66 patients (41 men, 25 women; 16 66 years) with double positivity for wasp and bee venoms were tested in the FEIA inhibition test in order to distinguish true double sensitization from cross reactivity. Patients were tested for the presence of IgE antibodies specific to oilseed rape (OSR) pollen and MUXF3 allergens, both of which are rich in cross reacting carbohydrate epitopes. RESULTS: Inhibition tests revealed true double sensitization in 37 patients (56.1%) and cross-reactivity in 29. Among those showing cross-reactivity, five were sensitized to honey bee venom and 24 to wasp venom. The median value of IgE specific for OSR pollen in patients sensitized to honey bee venom was 4.350 IU/ml, in patients sensitized to wasp venom 0.61 IU/ml, and in patients with double sensitization 0.25 IU/ml (P = 0.028, Kruskal-Wallis test). Findings for IgE specific for MUXF3 were similar. Discordance between OSR pollen positivity and MUXF3 positivity was found in 11.1% of the patients. CONCLUSION: The values of IgE specific for OSR pollen and MUXF3 in patients with primary sensitization to either honey bee venom or wasp venom were significantly higher than in patients with double sensitization. These results confirm that IgE antibodies against carbohydrates epitopes are a frequent cause of double positivity in tests for anti-venom IgE antibodies. PMID- 19562301 TI - The specificity of tests for anti-beta-lactam IgE antibodies declines progressively with increase of total serum IgE. AB - BACKGROUND: Immediate allergic reactions to beta-lactam antibiotics are mediated by specific IgE antibodies. The Phadia CAP System FEIA is a commercial method for quantification of specific IgE. We wished to determine anti-beta-lactam IgE antibodies in patients without penicillin allergy but with high levels of total IgE. METHODS: Sera from 41 patients (31 with high total IgE, 10 with low total IgE) were analyzed for IgE antibodies specific to penicilloyl G, penicilloyl V, amoxicilloyl and ampicilloyl using the CAP FEIA((R)) method that was available up to 2006. Seven sera that tested positive were rechecked in a new improved system available after 2006. RESULTS: In patients without a history of penicillin allergy, the specificities of commercial tests for anti-beta-lactam IgE antibodies were 100%, 60%, 27% and 20% at total IgE levels of 8-263 kU/l, 500-664 kU/l, 1000-2000 kU/l and > 2000 kU/l, respectively. In seven retested sera, only 2 (28%) were still positive for penicillin-specific IgE antibody. CONCLUSION: Before 2006, tests for anti-beta-lactam IgE antibody in patients with total IgE > 500 kU/l were probably often false positive. Patients who were diagnosed as penicillin allergic before 2006 solely on the basis of a positive CAP FEIA test for specific IgE should be considered for diagnostic reevaluation. PMID- 19562302 TI - The local and systemic side-effects of venom and inhaled-allergen subcutaneous immunotherapy. AB - BACKGROUND: Although immunotherapy is effective in allergic rhinitis, conjunctivitis, asthma and stinging insect hypersensitivity, it carries a risk of anaphylactic reactions. METHODS: In a 4-year retrospective survey, we investigated 1257 adult patients who had received venom or inhaled-allergen subcutaneous immunotherapy. The dose-increase phase was performed as the 2-day rush protocol for venom immunotherapy and the 6-week protocol for inhaled allergen immunotherapy. RESULTS: A total of 904 patients received venom immunotherapy and 353 patients inhaled-allergen immunotherapy. The prevalence of systemic reactions was 13.6%. The frequency of systemic reactions was higher during the maintenance phase than in the dose-increase phase (9.6% vs. 5.9%) and was highest in both phases of treatment with honeybee venom (P < 0.001). The majority of systemic reactions were mild. Five (0.4%) patients had reaction with a fall of blood pressure and were treated with adrenaline. There was no fatal outcome. The systemic side-effects during the dose-increase phase of venom immunotherapy occurred at a median dose of 46 microg (range 2-100 microg). Large local reactions occurred in 13.9% of patients without any significant difference between the allergens. CONCLUSIONS: We have shown that systemic reactions are not rare even during maintenance phase in patients with a well tolerated dose increase phase of treatment. The most prominent risk factor for systemic reactions was immunotherapy with honeybee extract. PMID- 19562316 TI - The interaction between ischemia-reperfusion and immune responses in the kidney. AB - Kidney ischemia-reperfusion injury (IRI) engages both the innate and adaptive immune responses. Cellular mediators of immunity, such as dendritic cells, neutrophils, macrophages, natural killer T, T, and B cells, contribute to the pathogenesis of renal injury after IRI. Postischemic kidneys express increased levels of adhesion molecules on endothelial cells and toll-like receptors on tubular epithelial cells. Soluble components of the immune system, such as complement activation proteins and cytokines, also participate in injury/repair of postischemic kidneys. Experimental studies on the immune response in kidney IRI have resulted in better understanding of the mechanisms underlying IRI and led to the discovery of novel therapeutic and diagnostic targets. PMID- 19562317 TI - Bumblebees exhibit the memory spacing effect. AB - Associative learning is key to how bees recognize and return to rewarding floral resources. It thus plays a major role in pollinator floral constancy and plant gene flow. Honeybees are the primary model for pollinator associative learning, but bumblebees play an important ecological role in a wider range of habitats, and their associative learning abilities are less well understood. We assayed learning with the proboscis extension reflex (PER), using a novel method for restraining bees (capsules) designed to improve bumblebee learning. We present the first results demonstrating that bumblebees exhibit the memory spacing effect. They improve their associative learning of odor and nectar reward by exhibiting increased memory acquisition, a component of long-term memory formation, when the time interval between rewarding trials is increased. Bombus impatiens forager memory acquisition (average discrimination index values) improved by 129% and 65% at inter-trial intervals (ITI) of 5 and 3 min, respectively, as compared to an ITI of 1 min. Memory acquisition rate also increased with increasing ITI. Encapsulation significantly increases olfactory memory acquisition. Ten times more foragers exhibited at least one PER response during training in capsules as compared to traditional PER harnesses. Thus, a novel conditioning assay, encapsulation, enabled us to improve bumblebee-learning acquisition and demonstrate that spaced learning results in better memory consolidation. Such spaced learning likely plays a role in forming long-term memories of rewarding floral resources. PMID- 19562318 TI - A morphological and life history comparison between desert populations of a sit and-pursue antlion, in reference to a co-occurring pit-building antlion. AB - Although most antlion species do not construct pits, the vast majority of studies on antlions focused on pit-building species. We report here on a transplant experiment aiming to test for morphological and life history differences between two desert populations of a sit-and-pursue antlion species, Lopezus fedtschenkoi (Neuroptera: Myrmeleontidae), originating from habitats, which mainly differ in plant cover and productivity. We raised the antlion larvae in environmental chambers simulating either hyper-arid or Mediterranean climate. We found significant differences in the morphology and life history of L. fedtschenkoi larvae between the two populations. For example, the larvae originating from the more productive habitat pupated faster and had a higher growth rate. In agreement with the temperature-size rule, antlions reached higher final mass in the colder Mediterranean climate and exhibited a higher growth rate, but there was no difference in their developmental time. Observed differences in morphology between populations as well as those triggered by climate growing conditions could be explained by differences in size allometry. We also provide a quantitative description of the allometric growth axis, based on 12 morphological traits. Comparing the responses of L. fedtschenkoi with those observed in a co occurring pit-building antlion indicated that there were neither shape differences that are independent of size nor was there a difference in the plasticity level between the two species. PMID- 19562319 TI - [The exact representation of skull base tumors as the basis for therapy]. PMID- 19562320 TI - [Adjuvant autologous tumour cell vaccination in patients with renal cell carcinoma. Overall survival analysis with a follow-up period in excess of more than 10 years]. AB - BACKGROUND: Organ-confined renal cell carcinoma (RCC) is associated with tumour progression after surgical therapy in approximately 30% of cases. However, of all recently available adjuvant treatment options, only the autologous tumour cell lysate vaccination therapy (Reniale) has been able to demonstrate a significant positive impact on progression-free survival in a phase III trial. Nevertheless, this therapeutic option has not yet been established as a standard adjuvant treatment. MATERIALS AND METHODS: Between August 1993 and December 1996, a total of 1,267 patients who underwent radical tumour nephrectomy at 84 German centres received Reniale outside a controlled trial. Of these patients, 692 presented at stage pT2-3, pNx-2, M0 (based on the 4th version of TNM classification). These patients were matched with a cohort of 861 patients not receiving any adjuvant treatment who underwent surgical therapy for RCC in a 15-year period in the Carl Thiem-Klinikum in Cottbus, Germany. Matching criteria included age, gender, pT stage, pN stage, grading, histological cell type, and UICC stage. This resulted in 495 matched pairs (study group n=990) that were comparable regarding demographic and tumour-specific criteria. Statistical analyses included univariate and multivariate analyses of overall survival (OS). Median follow-up time of all patients still alive at the end of the trial (n=667) was 11 years. RESULTS: In the vaccine group, OS after 5 and 10 years was 80.6% and 68.9%, respectively, whereas control patients had an OS of 79.2% and 62.1%, respectively (p=0.066). The 5-year OS of patients with pT3 RCC was 71.3% after vaccination therapy and 65.4% for control patients. After 10 years, 53.6% of the patients in the vaccine group and 36.2% in the control group were still alive (p=0.022). Median survival of patients with pT3 RCC was 81 months (SD 7.8) in the control group. This period was not achieved in the vaccine group. Multivariate Cox analysis revealed a significant positive impact of Reniale on OS among the whole study group [hazard ratio (HR) 1.28, p=0.030]. The analysis of patient subgroups showed a significant positive influence of Reniale for patients presenting with pT3 tumours (HR 1.67, p=0.001). CONCLUSION: Adjuvant postsurgical treatment with Reniale in patients presenting with stage pT3 RCC results in a significant enhancement of OS and should be considered especially in this group of patients. Further clinical trials integrating the recent TNM classification and comprising different risk constellations should follow in order to ultimately assess the value of adjuvant treatment with vaccination immunotherapy. PMID- 19562321 TI - Candida pneumonia in the ICU: myth or reality? PMID- 19562322 TI - Tight glycemic control and computerized decision-support systems: a systematic review. AB - OBJECTIVE: To identify and summarize characteristics of computerized decision support systems (CDSS) for tight glycemic control (TGC) and to review their effects on the quality of the TGC process in critically ill patients. METHODS: We searched Medline (1950-2008) and included studies on critically ill adult patients that reported original data from a clinical trial or observational study with a main objective of evaluating a given TGC protocol with a CDSS. RESULTS: Seventeen articles met the inclusion criteria. Eleven out of seventeen studies evaluated the effect of a new TGC protocol that was introduced simultaneously with a CDSS implementation. Most of the reported CDSSs were stand-alone, were not integrated in any other clinical information systems and used the "passive" mode requiring the clinician to ask for advice. Different implementation sites, target users, and time of advice were used, depending on local circumstances. All controlled studies reported on at least one quality indicator of the blood glucose regulatory process that was improved by introducing the CDSS. Nine out of ten controlled studies either did not report on the number of hypoglycemia events (one study), or reported on no change (six studies) or even a reduction in this number (two studies). CONCLUSIONS: While most studies evaluating the effect of CDSS on the quality of the TGC process found improvement when evaluated on the basis of the quality indicators used, it is impossible to define the exact success factors, because of simultaneous implementation of the CDSS with a new or modified TGC protocol and the hybrid solutions used to integrate the CDSS into the clinical workflow. PMID- 19562324 TI - Identification of a novel M-superfamily conotoxin with the ability to enhance tetrodotoxin sensitive sodium currents. AB - In this work, a novel M-superfamily conotoxin, designated lt3a, was purified from the crude venom of Conus litteratus. Combined with peptide sequencing, MALDI-TOF mass spectrometry and cDNA cloning techniques, the amino acid sequence of lt3a was supposed to be DgammaCCgamma OQWCDGACDCCS, where O is hydroxyproline and gamma is carboxyglutamate. The Cys framework of lt3a (-CC-C-C-CC-) is similar to that of psi-, mu-, kappaM-conotoxins, which are representatives of M-conotoxins. Peptide lt3a is categorized into M1 branch based on the number of residues in the last Cys loop. Whole cell patch-clamp study on adult rat dorsal root ganglion neurons indicated that lt3a could enhance tetrodotoxin-sensitive sodium currents. This is a previously unknown function of M-superfamily conotoxins. PMID- 19562323 TI - Safety and efficacy of ketorolac in children after cardiac surgery. AB - OBJECTIVE: To evaluate the nephrotoxic and opioid-sparing effects of ketorolac in children after cardiac surgery. DESIGN: A retrospective cohort study. SETTING: A Cardiac Critical Care Unit in a university-affiliated children's hospital. SUBJECTS: Children less than 18 years of age who underwent low-risk cardiac surgery from July 2002 to December 2005. RESULTS: Among 248 children studied, 108 received ketorolac and 140 did not. The ketorolac group was older, included a larger proportion of atrial septum defect repairs and a smaller proportion of ventricular septum defect repairs compared to the control group. The median change in serum creatinine did not differ between the ketorolac group and the control group (% change [IQR]); 12% [1-25] increase versus 12% [-3 to 31] increase, P = 0.86. On postoperative day 0 or 1, the ketorolac group received less opioids than control group. There was no difference in duration of mechanical ventilation or in length of stay between groups. CONCLUSION: Ketorolac started in the first 12 h after a low-risk cardiac surgery in children is not associated with a measurable difference in renal function. The data suggest that ketorolac may be effective in reducing the exposure to opioids. Further studies are required to define subsets of children after cardiac surgery who could safely benefit from ketorolac therapy to reduce pain. PMID- 19562325 TI - In vitro amyloid Abeta(1-42) peptide aggregation monitoring by asymmetrical flow field-flow fractionation with multi-angle light scattering detection. AB - Self-assembly of the 42-amino-acid-long amyloid peptide Abeta(1-42) into insoluble fibrillar deposits in the brain is a crucial event in the pathogenesis of Alzheimer's disease. The fibril deposition occurs through an aggregation process during which transient and metastable oligomeric intermediates are intrinsically difficult to be accurately monitored and characterised. In this work, the time-dependent Abeta(1-42) aggregation pattern is studied by asymmetrical flow field-flow fractionation with on-line multi-angle light scattering detection. This technique allows separating and obtaining information on the molar mass (M(r)) and size distribution of both the early-forming soluble aggregates and the late prefibrillar and fibrillar species, the latter having very high M(r). Preliminary results demonstrate that unique information on the dynamic aggregation process can be obtained, namely on the M(r) and size of the forming aggregates as well as on their formation kinetics. PMID- 19562326 TI - Ion mobility-mass spectrometry analysis of isomeric carbohydrate precursor ions. AB - The rapid separation of isomeric precursor ions of oligosaccharides prior to their analysis by mass spectrometry to the nth power (MS(n)) was demonstrated using an ambient pressure ion mobility spectrometer (IMS) interfaced with a quadrupole ion trap. Separations were not limited to specific types of isomers; representative isomers differing solely in the stereochemistry of sugars, in their anomeric configurations, and in their overall branching patterns and linkage positions could be resolved in the millisecond time frame. Physical separation of precursor ions permitted independent mass spectra of individual oligosaccharide isomers to be acquired to at least MS(3), the number of stages of dissociation limited only practically by the abundance of specific product ions. IMS-MS(n) analysis was particularly valuable in the evaluation of isomeric oligosaccharides that yielded identical sets of product ions in tandem mass spectrometry experiments, revealing pairs of isomers that would otherwise not be known to be present in a mixture if evaluated solely by MS dissociation methods alone. A practical example of IMS-MS(n) analysis of a set of isomers included within a single high-performance liquid chromatography fraction of oligosaccharides released from bovine submaxillary mucin is described. PMID- 19562328 TI - Disentangling perceptual from motor implicit sequence learning with a serial color-matching task. AB - This paper contributes to the domain of implicit sequence learning by presenting a new version of the serial reaction time (SRT) task that allows unambiguously separating perceptual from motor learning. Participants matched the colors of three small squares with the color of a subsequently presented large target square. An identical sequential structure was tied to the colors of the target square (perceptual version, Experiment 1) or to the manual responses (motor version, Experiment 2). Short blocks of sequenced and randomized trials alternated and hence provided a continuous monitoring of the learning process. Reaction time measurements demonstrated clear evidence of independently learning perceptual and motor serial information, though revealed different time courses between both learning processes. No explicit awareness of the serial structure was needed for either of the two types of learning to occur. The paradigm introduced in this paper evidenced that perceptual learning can occur with SRT measurements and opens important perspectives for future imaging studies to answer the ongoing question, which brain areas are involved in the implicit learning of modality specific (motor vs. perceptual) or general serial order. PMID- 19562329 TI - Effect of ritonavir on the pharmacokinetics of the benzimidazoles albendazole and mebendazole: an interaction study in healthy volunteers. AB - BACKGROUND: Benzimidazoles are often used concomitantly with protease inhibitors in patients with helminthic disease and HIV infection. Low bioavailability and extensive first-pass metabolism make benzimidazoles prone to pharmacokinetic drug interactions. The aim of the present study was to investigate potential drug interactions between the benzimidazoles albendazole and mebendazole and the potent CYP3A4 inhibitor ritonavir. METHODS: Sixteen healthy volunteers were administered a single oral dose of 1,000 mg mebendazole or 400 mg albendazole (2 x n = 8). AUC, C(max), and t(1/2) of mebendazole, albendazole, and albendazole sulfoxide were studied in absence and after short-term (2 doses) and long-term (8 days) treatment with ritonavir 200 mg bid. RESULTS: Pharmacokinetic parameters of albendazole and mebendazole were not changed by short-term administration of ritonavir. However, long-term administration of ritonavir resulted in significant changes in albendazole and mebendazole disposition, with a significant decrease in AUC(0-24) (27 and 43% of baseline for albendazole and mebendazole, respectively) and C(max) (26 and 41% of baseline, respectively). CONCLUSION: The AUC(0-24) of benzimidazoles decreased after long-term use of ritonavir, while no changes in pharmacokinetic profiles were observed under short-term administration. These findings might help to optimize benzimidazole efficacy when used in combination with protease inhibitors. PMID- 19562332 TI - Lambda exonuclease digestion of CGG trinucleotide repeats. AB - Fragile X syndrome and other trinucleotide diseases are characterized by an elongation of a repeating DNA triplet. The ensemble-averaged lambda exonuclease digestion rate of different substrates, including one with an elongated FMR1 gene containing 120 CGG repeats, was measured using absorption and fluorescence spectroscopy. By use of magnetic tweezers sequence-dependent digestion rates and pausing was measured for individual lambda exonucleases. Within the triplet repeats a lower average and narrower distribution of rates and a higher frequency of pausing was observed. PMID- 19562330 TI - Endovascular transplantation of stem cells to the injured rat CNS. AB - INTRODUCTION: Transplantation procedures using intraparenchymal injection of stem cells result in tissue injury in addition to associated surgical risks. Intravenous injection of mesenchymal stem cells gives engraftment to lesions, but the method has low efficiency and specificity. In traumatic brain injuries (TBI), there is a transient breakdown of the blood-brain barrier and an inflammatory response, which increase migration of cells from blood to parenchyma. The aim of this investigation was to analyze the effect of intra-arterial administration on cellular engraftment. METHODS: Experimental TBI was produced in a rat model. Endovascular technique was used to administer human mesenchymal stem cells in the ipsilateral internal carotid artery. Evaluation of engraftment and side effects were performed by immunohistochemical analysis of the brain and several other organs. The results were compared to intravenous administration of stem cells. RESULTS: Intra-arterial transplantation of mesenchymal stem cells resulted in central nervous system (CNS) engraftment without thromboembolic ischemia. We observed a significantly higher number of transplanted cells in the injured hemisphere after intra-arterial compared to intravenous administration both 1 day (p < 0.01) and 5 days (p < 0.05) after the transplantation. Some cells were also detected in the spleen but not in the other organs analyzed. CONCLUSION: Selective intra-arterial administration of mesenchymal stem cells to the injured CNS is a minimally invasive method for transplantation. The method is significantly more efficient than the intravenous route and causes no side effects in the current model. The technique can potentially be used for repeated transplantation to the CNS after TBI and in other diseases. PMID- 19562333 TI - Development of an enzyme activity screening system for beta-glucosidase displaying yeasts using calcium alginate micro-beads and flow sorting. AB - Recent reports on high-speed affinity screening systems for yeast cells using flow cytometry have not been adapted to screening yeast cells that display hydrolyzing enzymes, since the fluorescent molecules which are released from fluoresceinated substrate diffuse into solution after enzymatic reaction. In this research, yeast cells displaying beta-glycosidase were individually captured in micro-sized calcium alginate beads by using the newly developed reverse micelle method to prevent diffusion of hydrolyzed fluorescent substrates. By adopting flow sorting to these captured cells, active cells were successfully enriched about 82-fold from a mixed suspension with negative controls. This system should be a useful method for high-speed screening of yeast cells that display various hydrolyzing enzymes and has potential application to screening randomized libraries of enzyme-displayed yeast cells with higher activities. PMID- 19562334 TI - Stimulation of artemisinin synthesis by combined cerebroside and nitric oxide elicitation in Artemisia annua hairy roots. AB - This work examined the accumulation of artemisinin and related secondary metabolism pathways in hairy root cultures of Artemisia annua L. induced by a fungal-derived cerebroside (2S,2'R,3R,3'E,4E,8E)-1-O-beta-D-glucopyranosyl-2-N (2'-hydroxy-3'-octadecenoyl)-3-hydroxy-9-methyl-4,8-sphingadienine. The presence of the cerebroside induced nitric oxide (NO) burst and artemisinin biosynthesis in the hairy roots. The endogenous NO generation was examined to be involved in the cerebroside-induced biosynthesis of artemisinin by using NO inhibitors, N (omega)-nitro-L-arginine methyl ester and 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide. The gene expression and activity of 3 hydroxy-3-methylglutaryl CoA reductase and 1-deoxy-D-xylulose 5-phosphate synthase were stimulated by the cerebroside, but more strongly by the potentiation of NO. While the mevalonate pathway inhibitor, mevinolin, only partially inhibited the induced artemisinin accumulation, the plastidic 2-C methyl-D-erythritol 4-phosphate pathway inhibitor, fosmidomycin, nearly arrested artemisinin accumulation induced by cerebroside and the combination elicitation with an NO donor, sodium nitroprusside (SNP). With the potentiation by SNP at 10 microM, the cerebroside elicitor stimulated artemisinin production in 20-day-old hairy root cultures up to 22.4 mg/l, a 2.3-fold increase over the control. These results suggest that cerebroside plays as a novel elicitor and the involvement of NO in the signaling pathway of the elicitor activity for artemisinin biosynthesis. PMID- 19562335 TI - Generation of chromosomal DNA during alkaline lysis and removal by reverse micellar extraction. AB - The separation of structurally related impurities from pharmaceutical plasmid DNA by highly scalable purification techniques is a challenge for biochemical engineering. Next to RNA, proteins, and lipopolysaccharides, the chromosomal DNA of the plasmid replicating host has to be removed. Here, we describe the application of reverse micellar extraction for the separation of chromosomal from plasmid DNA. By applying different procedures for alkaline lysis, bacterial lysates with different amounts of chromosomal DNA were generated. A reverse micellar extraction step enabled us to deplete the concentration of this impurity below the required level of 50 mg g(-1) of plasmid DNA with almost complete plasmid recovery. PMID- 19562337 TI - Focal fat mimicking multiple hepatic metastases on FDG PET/CT imaging. PMID- 19562336 TI - EANM guidelines for ventilation/perfusion scintigraphy : Part 1. Pulmonary imaging with ventilation/perfusion single photon emission tomography. AB - Pulmonary embolism (PE) can only be diagnosed with imaging techniques, which in practice is performed using ventilation/perfusion scintigraphy (V/P(SCAN)) or multidetector computed tomography of the pulmonary arteries (MDCT). The epidemiology, natural history, pathophysiology and clinical presentation of PE are briefly reviewed. The primary objective of Part 1 of the Task Group's report was to develop a methodological approach to and interpretation criteria for PE. The basic principle for the diagnosis of PE based upon V/P(SCAN) is to recognize lung segments or subsegments without perfusion but preserved ventilation, i.e. mismatch. Ventilation studies are in general performed after inhalation of Krypton or technetium-labelled aerosol of diethylene triamine pentaacetic acid (DTPA) or Technegas. Perfusion studies are performed after intravenous injection of macroaggregated human albumin. Radiation exposure using documented isotope doses is 1.2-2 mSv. Planar and tomographic techniques (V/P(PLANAR) and V/P(SPECT)) are analysed. V/P(SPECT) has higher sensitivity and specificity than V/P(PLANAR). The interpretation of either V/P(PLANAR) or V/P(SPECT) should follow holistic principles rather than obsolete probabilistic rules. PE should be reported when mismatch of more than one subsegment is found. For the diagnosis of chronic PE, V/P(SCAN) is of value. The additional diagnostic yield from V/P(SCAN) includes chronic obstructive lung disease (COPD), heart failure and pneumonia. Pitfalls in V/P(SCAN) interpretation are considered. V/P(SPECT) is strongly preferred to V/P(PLANAR) as the former permits the accurate diagnosis of PE even in the presence of comorbid diseases such as COPD and pneumonia. Technegas is preferred to DTPA in patients with COPD. PMID- 19562339 TI - Expression of CD13/aminopeptidase N in precursor B-cell leukemia: role in growth regulation of B cells. AB - Expression of cell surface CD13 in acute B-cell leukemia (ALL-B) is often viewed, as an aberrant expression of a myeloid lineage marker. Here, we attempted to study the stage specific expression of CD13 on ALL-B blasts and understand its role in leukemogenesis as pertaining to stage of B-cell ontogeny. A total of 355 cases of different hematological malignancies were diagnosed by immunophenotyping. Among 68 cases of early B-cell ALL, 22 cases with distinct immunophenotype was identified as immature B-cell ALL. Blasts from these ALL-B patients demonstrated prominent expression of CD10, CD19, CD22, but neither cytoplasmic nor surface IgM receptors. This strongly indicates leukemogenesis at an early stage of B-cell development. We also identified, the existence of a subpopulation of cells with remarkably similar phenotype in non-leukemic marrow from healthy subjects (expressing CD10, CD19, CD22, CD24, Tdt together with the co-expression of CD13). This sub-population of B cells concomitantly expressing CD13 appeared to be a highly proliferating group. By blocking their cell surface CD13 in leukemic blasts with monoclonal antibody we were able to inhibit their proliferation. We hypothesized that neoplastic transformation at this stage may be facilitated by CD13. CD13 may thus be an important target for novel molecular therapy of early stage acute B-cell leukemia. PMID- 19562338 TI - A systematic review of humoral immune responses against tumor antigens. AB - This review summarizes studies on humoral immune responses against tumor associated antigens (TAAs) with a focus on antibody frequencies and the potential diagnostic, prognostic, and etiologic relevance of antibodies against TAAs. We performed a systematic literature search in Medline and identified 3,619 articles on humoral immune responses and TAAs. In 145 studies, meeting the inclusion criteria, humoral immune responses in cancer patients have been analyzed against over 100 different TAAs. The most frequently analyzed antigens were p53, MUC1, NY ESO-1, c-myc, survivin, p62, cyclin B1, and Her2/neu. Antibodies against these TAAs were detected in 0-69% (median 14%) of analyzed tumor patients. Antibody frequencies were generally very low in healthy individuals, with the exception of few TAAs, especially MUC1. For several TAAs, including p53, Her2/neu, and NY-ESO 1, higher antibody frequencies were reported when tumors expressed the respective TAA. Antibodies against MUC1 were associated with a favorable prognosis while antibodies against p53 were associated with poor disease outcome. These data suggest different functional roles of endogenous antibodies against TAAs. Although data on prediagnostic antibody levels are scarce and antibody frequencies for most TAAs are at levels precluding use in diagnostic assays for cancer early detection, there is some promising data on achieving higher sensitivity for cancer detection using panels of TAAs. PMID- 19562340 TI - Identification of a 17beta-hydroxysteroid dehydrogenase type 12 pseudogene as the source of a highly restricted BALB/c Meth A tumor rejection peptide. AB - Mass spectrometric analysis identified the peptide recognized by a cytotoxic T lymphocyte (CTL) specific for the chemically induced BALB/c Meth A sarcoma as derived from a 17beta-hydroxysteroid dehydrogenase type 12 (Hsd17b12) pseudogene present in the BALB/c genome, but only expressed in Meth A sarcoma. The sequence of the peptide is TYDKIKTGL and corresponds to Hsd17b12(114-122) with threonine instead of isoleucine at codon 114 and is designated Hsd17b12(114T). Immunization of mice with an Hsd17b12(114T) peptide-pulsed dendritic cell-based vaccine or a non-viral plasmid construct expressing the Hsd17b12(114T) peptide protected the mice from lethal Meth A tumor challenge in tumor rejection assays. A Hsd17b12(114 122) peptide-pulsed vaccine was ineffective in inducing resistance in mice to Meth A sarcoma. These results confirm the immunogenicity of the identified tumor peptide, as well as demonstrate the efficacies of these vaccine vehicles. These findings suggest that the role of the human homolog of Hsd17b12, HSD17B12, as a potential human tumor antigen be explored. PMID- 19562341 TI - Lenalidomide in 5q minus myelodysplastic syndrome: how long is enough? PMID- 19562343 TI - A comparison of quality of life, demographic and clinical characteristics of Brazilian men with fibromyalgia syndrome with male patients with depression. AB - The primary aim of this study was to assess the quality of life (Qol) in men with fibromyalgia (FM) as compared to the Qol of depressive patients, using the SF-36 questionnaire. The secondary objectives were as follows: to describe demographic and clinical characteristics of male patients with FM; to evaluate basal levels of dehydroepiandrosterone sulfate (DHEAS) and total testosterone in both groups; and to investigate the relationship among pain, tender points, anxiety, and depression in these patients. Fifty men with FM and 20 depressed males, matched by age and body mass index entered the study. All participants answered the SF 36, the Beck Depression Inventory and the State-Trait Anxiety Inventory questionnaire. Radioimmunoassay was used to determine serum concentration of DHEAS and total testosterone. Patients and controls had similar demographic characteristics. The scores were significantly lower in all domains of the SF-36 in patients with FM as compared with the depressive controls. No significant differences were observed among patient and control in the mean concentration of either DHEAS or total testosterone. Male patients with FM experience worse Qol than depressive men. Depression was influential on mental health, whereas FM impacted on both physical and mental health. PMID- 19562342 TI - Ccm1p/Ygr150cp, a pentatricopeptide repeat protein, is essential to remove the fourth intron of both COB and COX1 pre-mRNAs in Saccharomyces cerevisiae. AB - YGR150C gene product (Ygr150cp) is one of the three mitochondrially located Saccharomyces cerevisiae proteins with pentatricopeptide repeat (PPR) motifs. Ygr150cp is essential for mitochondrial functionality but its molecular targets are still unknown. This study was undertaken to define the role of Ygr150cp in mitochondria biogenesis. Repression of Ygr150cp expression in complemented mutants prevented their use of glycerol or lactate, but allowed limited growth on ethanol-containing medium. RNA hybridization studies showed that Deltaygr150c meiotic segregants produced COB and COX1 transcripts but failed to process them into the mature forms. Detailed RT-PCR assays revealed that Deltaygr150c specifically failed to remove the fourth intron of both COB and COX1 pre-mRNAs while all other group I introns were excised. Expression of Ygr150cp mutants without any of the PPR motifs did not complement the growth phenotype. Accordingly, we designate YGR150C as CCM1 (COB and COX1 mRNA maturation). This report provides the first evidence of PPR protein involvement in the specific removal of group I introns in mitochondria of S. cerevisiae. PMID- 19562345 TI - Analysis of gene expression profile in pollen development of recessive genic male sterile Brassica napus L. line S45A. AB - Male sterility in a near-isogenic line S45AB after 25 generations of subcrossing is controlled by two pairs of duplicate genes. The genotype of S45A is Bnms1Bnms1Bnms2Bnms2, and that of S45B is BnMs1Bnms1Bnms2Bnms2, respectively. Histological observations revealed that abnormal anther development appeared in the tapetum and pollen exine during the tetrad stage. This male sterility was characterized by hypertrophy of the tapetal cells at the tetrad stage and a complete lack of microspore exine after the release of microspores from the tetrads. To elucidate the mechanism of this recessive genic male sterility, the flower bud expression profiles of the S45A and S45B lines were analyzed using an Arabidopsis thaliana ATH1 oligonucleotide array. When compared with the S45B line, 69 genes were significantly downregulated, and 46 genes were significantly upregulated in the S45A line. Real-time polymerase chain reaction (PCR) was then used to verify the results of the microarray analysis, and the majority of the downregulated genes in the S45A line were abundantly and specifically expressed in the anther. The results of the real-time PCR suggest that Bnms1 might be involved in the metabolism of lipid/fatty acids, and the homologous mutation of Bnms1 may either block the biosynthesis of sporopollenin or block sporopollenin from being deposited on the microspore surface, thus, preventing pollen exine formation. The role of Bnms1 in the regulatory network of exine formation is also discussed as well. PMID- 19562346 TI - The value of an artificial neural network in the decision-making for prostate biopsies. AB - PURPOSE: In majority of patients who are subjected to prostate biopsies, no prostate cancer (PCa) is found. It is important to prevent unnecessary biopsies since serious complications may occur. An artificial neural network (ANN) may be able to predict the risk of the presence of PCa. METHODS: Included were all patients, who underwent transrectal ultrasound-guided prostate biopsies between June 2006 and June 2007 with a total PSA (tPSA) level between 2 and 20 microg/l. The patients were divided into two groups according to their tPSA level (2-10 microg/l and 10-20 microg/l). The ANN Prostataclass of the Universitatsklinikum Charite in Berlin was used. The predictions of the ANN were compared to the pathology results of the biopsies. RESULTS: Overall 165 patients were included. PCa was diagnosed in 53 patients, whereas the ANN predicted "no risk" in 19 of these patients (36%). The ANN output receiver operator characteristic (ROC) plots for the range of tPSA 2-10 microg/l and tPSA 10-20 microg/l showed an area under the curve (AUC) of 63 and 88% for the initial biopsy group, versus 69 and 57%, respectively, for the repeat biopsy group. CONCLUSIONS: The ANN resulted in a false negative rate of 36%, missing PCa in 19 patients. For use in an outpatient clinical setting, this ANN is insufficient to predict the risk of presence of PCa reliably. PMID- 19562344 TI - A systematic MEDLINE analysis of therapeutic approaches in ankylosing spondylitis. AB - Ankylosing spondylitis (AS) is a chronic inflammatory disorder involving the sacroiliac joints (SIJs), spine and less frequently the peripheral joints. Traditionally, it is well recognised that AS is a challenging disease to manage due to the lack of effective therapeutic options. Current evidence would suggest this has changed and there are now a number of therapies available that provide persistent control of inflammatory symptoms with improvement in daily function. NSAIDs remain the first step in patient treatment. Sulphasalazine may be effective in peripheral arthritis and there are emerging data to support its use in early inflammatory back pain. Studies have shown that pamidronate and steroid injection into SIJ have a symptom-modifying effect in AS. Current data suggest that anti-TNF treatment promises early benefit which is likely to continue in the longer term. Treatment with biologics should be considered sooner rather than later in the management of AS. PMID- 19562347 TI - The f/t-PSA ratio in diagnosis of in-patients and out-patients: a unitary cutoff value is not useful! AB - PURPOSE: In the prostate specific antigen (PSA) range of 4-10 ng/ml after a negative digital rectal examination, the PSA value indicates a lack of specificity with a carcinoma detection rate of roughly 20%. To improve the biopsy/carcinoma ratio, the interdisciplinary consensus recommends free PSA (fPSA) measurement. This does not take into account the pre-analysis when the cutoff value is established for biopsy indication. METHODS: In the present study, an in-patient cohort whose blood samples were immediately analysed was compared with an out-patient cohort whose sample processing was delayed by between 24 and 48 h. RESULTS: The in-patient cohort comprises 382 patients with 99 prostate carcinomas, the out-patient cohort 987 patients with 235 carcinomas. In the in patient cohort a sensitivity of 90% with a cutoff value of 25% for the f/t-PSA ratios is achieved with the theoretical possibility of reducing the number of punch biopsies by 34.6%. A sensitivity of 90% in the out-patient cohort necessitates a cutoff value of 18% for the f/t-PSA ratios. The specificity is 35.3% with a possible biopsy reduction of 29.1%. CONCLUSIONS: The cutoff values from cohorts with an immediate fPSA measurement cannot be adopted for a typical out-patient cohort whose analyses are delayed. On the contrary, an individual adjustment is necessary which corresponds to the pre-analysis. PMID- 19562348 TI - Vascular risk factors and their association to serum androgen levels in a population-based cohort of 75-year-old men over 5 years: results of the VITA study. AB - PURPOSE: In contrast to the high prevalence of late onset hypogonadism, little is known regarding correlates for low androgen levels in aging men. METHODS: We investigated participants of the Vienna Transdanube Aging study and assessed the relationship between vascular risk factors and hormonal status over 5 years. RESULTS: A total of 247 men with a mean age of 75.8 years were analyzed. Despite a tendency for lower testosterone levels in men with vascular risk factors and vascular diseases, none of these associations reached statistical significance. Men with low DHEA-S levels had a lower risk of hypercholesterinemia (-55.2%; P = 0.01) yet an increased prevalence of diabetes (+95.7%; P = 0.02) and coronary heart disease (+47.6%; P = 0.05). Testosterone and DHEA-S remained stable over 5 years of follow-up. CONCLUSION: While reduced levels of total testosterone did not show an association to vascular disease, low DHEA-S was linked to hypercholesterinemia, diabetes, and coronary heart disease. PMID- 19562349 TI - The difference in oxidative stress of the blood between using 5% glucose water and distilled water as the irrigant for BPH patients undergoing transurethral resection of the prostate. AB - INTRODUCTION: To evaluate the difference in oxidative stress in the blood between using 5% glucose water and using distilled water as the irrigant for BPH patients receiving transurethral resection of the prostate (TURP), we conducted this prospective study, on a total of 38 patients with symptomatic BPH. MATERIALS AND METHODS: All the patients were randomly divided into 2 groups. Distilled water in group A and 5% glucose water in group B were used as the irrigant. The oxidative stress and antioxidant capacity were evaluated by measuring malondialdehyde (MDA), mitochondrial DNA (mtDNA) copy number and total capacity of antioxidants (TOA) in the blood. The data were correlated with serum creatinine and sodium, which were measured before and immediately after TURP in all patients. RESULTS: Serum creatinine increased from 0.86 +/- 0.17 to 1.02 +/- 0.20 mg/dL while serum sodium decreased from 139.7 +/- 2.3 to 136.4 +/- 4.0 mEq/L immediately after surgery in patients of group A (p < 0.05). MDA was increased and TOA decreased in all the patients immediately after TURP. Patients in group A had significantly higher MDA, lower mtDNA copy number, higher degree of oxidative mtDNA damage and lower TOA than those in group B immediately after TURP. CONCLUSION: In conclusion, we observed an increase of MDA and decrease of TOA in blood in all the patients immediately after TURP, which might induce renal damage and decrease serum sodium. Moreover, the oxidative stress levels in blood of patients in group A were significantly higher than those in group B immediately after TURP. PMID- 19562350 TI - Inside the guts of wood-eating catfishes: can they digest wood? AB - To better understand the structure and function of the gastrointestinal (GI) tracts of wood-eating catfishes, the gross morphology, length, and microvilli surface area (MVSA) of the intestines of wild-caught Panaque nocturnus, P. cf. nigrolineatus "Maranon", and Hypostomus pyrineusi were measured, and contrasted against these same metrics of a closely related detritivore, Pterygoplichthys disjunctivus. All four species had anatomically unspecialized intestines with no kinks, valves, or ceca of any kind. The wood-eating catfishes had body size corrected intestinal lengths that were 35% shorter than the detritivore. The MVSA of all four species decreased distally in the intestine, indicating that nutrient absorption preferentially takes place in the proximal and mid-intestine, consistent with digestive enzyme activity and luminal carbohydrate profiles for these same species. Wild-caught Pt. disjunctivus, and P. nigrolineatus obtained via the aquarium trade, poorly digested wood cellulose (<33% digestibility) in laboratory feeding trials, lost weight when consuming wood, and passed stained wood through their digestive tracts in less than 4 h. Furthermore, no selective retention of small particles was observed in either species in any region of the gut. Collectively, these results corroborate digestive enzyme activity profiles and gastrointestinal fermentation levels in the fishes' GI tracts, suggesting that the wood-eating catfishes are not true xylivores such as beavers and termites, but rather, are detritivores like so many other fishes from the family Loricariidae. PMID- 19562351 TI - Ependymoma: lessons from the past, prospects for the future. PMID- 19562353 TI - Our impact factor of 5.31: why it increased and what it means to readers and authors. PMID- 19562352 TI - Risk management in pediatric surgery. AB - PURPOSE: To present the experience documented over 1 year of analysis and quality control on surgical complications and organizational accidents. METHODS: All children admitted during the study period at our Institution were included in the analysis, which consisted of four phases: (1) definition and standardization of perioperative diagnostic and therapeutic tracks; (2) staff education; (3) documentation and data implementation, and (4) "Morbidity and Mortality" audit. RESULTS: Over a 1-year study period, 3,116 children were admitted to our Institution: 2,222 out of 3,116 (71.3%) children underwent a surgical procedure. A total number of 184 complications were recorded in 149 patients. One hundred and seventy-one (92.9%) complications occurred following a surgical procedure. Fifty-six out of 149 complicated patients (37.6%) required a re-operation. Thirty five out of 184 (19%) complications were classified as organizational. Infection represented the most common complication. All cases of anastomotic dehiscence and perforation, bowel obstruction, and stoma malfunction required reintervention. None of the postoperative bleedings required a second surgical procedure. CONCLUSION: Although a proper statistical comparison with literature complication rates is not feasible, our experience confirms the importance of quality-control audit in health care systems. Prolonged observation, long-term follow up, and comparison with previous results will represent our future goal. PMID- 19562354 TI - Primary leptomeningeal oligodendroglioma with documented progression to anaplasia and t(1;19)(q10;p10) in a child. PMID- 19562356 TI - Early results in the treatment of proximal humeral fractures with a polyaxial locking plate. AB - OBJECTIVES: We report early results using a second generation locking plate, non contact bridging plate (NCB PH((R)), Zimmer Inc. Warsaw, IN, USA), for the treatment of proximal humeral fractures. The NCB PH combines conventional plating technique with polyaxial screw placement and angular stability. DESIGN: Prospective case series. SETTING: A single level-1 trauma center. PATIENTS: A total of 50 patients with proximal humeral fractures were treated from May 2004 to December 2005. INTERVENTION: Surgery was performed in open technique in all cases. MAIN OUTCOME MEASURES: Implant-related complications, clinical parameters (duration of surgery, range of motion, Constant-Murley Score, subjective patient satisfaction, complications) and radiographic evaluation [union, implant loosening, implant-related complications and avascular necrosis (AVN) of the humeral head] at 6, 12 and 24 weeks. RESULTS: All fractures available to follow up (48 of 50) went to union within the follow-up period of 6 months. One patient was lost to follow-up, one patient died of a cause unrelated to the trauma, four patients developed AVN with cutout, one patient had implant loosening, three patients experienced cutout and one patient had an axillary nerve lesion (onset unknown). The average age- and gender-related Constant Score (n = 35) was 76. CONCLUSIONS: The NCB PH combines conventional plating technique with polyaxial screw placement and angular stability. Although the complication rate was 19%, with a reoperation rate of 12%, the early results show that the NCB PH is a safe implant for the treatment of proximal humeral fractures. PMID- 19562357 TI - Spondylodiscitis of the lumbar spine in a non-immunocompromised host caused by Yersinia enterocolitica O:9. AB - Here presented is an extremely rare case of a spinal osteomyelitis (L5-S1) with epidural empyema in a non-immunocompromised 62-year-old man caused by Yersinia enterocolitica O:9. The infection occurred acutely and required immediate surgical treatment. Y. enterocolitica was cultured from the empyema fluid, wound swabs of the intervertebral disc L5-S1 and stool cultures. Following the surgical decompression and antibiotic treatment, the patient recovered completely, without neurological deficits. A review of the literature revealed only sparse cases of spondylodiscitis due to other Y. enterocolitica serogroups. To our knowledge, we report here the first case of a spondylodiscitis of the lumbar spine caused by Y. enterocolitica serovar O:9 in a non-immunocompromised patient. PMID- 19562355 TI - Neuronal loss in Pelizaeus-Merzbacher disease differs in various mutations of the proteolipid protein 1. AB - Mutations affecting proteolipid protein 1 (PLP1), the major protein in central nervous system myelin, cause the X-linked leukodystrophy Pelizaeus-Merzbacher disease (PMD). We describe the neuropathologic findings in a series of eight male PMD subjects with confirmed PLP1 mutations, including duplications, complete gene deletion, missense and exon-skipping. While PLP1 mutations have effects on oligodendrocytes that result in mutation-specific degrees of dysmyelination, our findings indicate that there are also unexpected effects in the central nervous system resulting in neuronal loss. Although length-dependent axonal degeneration has been described in PLP1 null mutations, there have been no reports on neuronal degeneration in PMD patients. We now demonstrate widespread neuronal loss in PMD. The patterns of neuronal loss appear to be dependent on the mutation type, suggesting selective vulnerability of neuronal populations that depends on the nature of the PLP1 disturbance. Nigral neurons, which were not affected in patients with either null or severe misfolding mutations, and thalamic neurons appear particularly vulnerable in PLP1 duplication and deletion patients, while hippocampal neuronal loss was prominent in a patient with complete PLP1 gene deletion. All subjects showed cerebellar neuronal loss. The patterns of neuronal involvement may explain some clinical findings, such as ataxia, being more prominent in PMD than in other leukodystrophies. While the precise pathogenetic mechanisms are not known, these observations suggest that defective glial functions contribute to neuronal pathology. PMID- 19562358 TI - Splenic artery: peak systolic velocity of normal fetuses. AB - OBJECTIVE: To establish a reference range of splenic artery peak systolic velocity 1(SpA-PSV) in the normal singleton pregnancies (14-40 weeks). METHODS: A prospective descriptive study was conducted on uncomplicated singleton pregnancies with normal fetuses and accurate gestational age were recruited into the study. The Doppler measurements of SpA-PSV were performed by the experienced sonographers with the high-resolution machine (Aloka Prosound alpha-10, Tokyo, Japan, or Voluson E8, GE Healthcare, USA). RESULTS: A total of 540 measurements were performed, ranging from 15 to 30 for each gestational week (GA). The best regression model between GA and SpA-PSV was observed to be linear function with an equation as follows: SpA-PSV (cm/s)=-1.433+1.186 (GA, weeks) (r2=0.573). The table of nomogram for various percentile ranges was constructed. The results show a continuous increase in SpA-PSV over the period from 14 to 40 weeks. CONCLUSION: A nomogram for SpA-PSV for each GA during 14-40 weeks was constructed. This reference range may be a useful non-invasive tool in risk assessment for fetal anemia, especially due to homozygous alpha-thalassemia-1 or fetal isoimmunization. PMID- 19562359 TI - Topographical correlations of lateral medullary infarction with caloric- and vestibular-evoked myogenic potential results. AB - This study investigated the correlation of caloric- and vestibular-evoked myogenic potential (VEMP) results with topographical lesions of lateral medullary infarction. Five patients with lateral medullary infarction were enrolled in this study. Each patient underwent a battery of tests, including audiometry, caloric test, VEMP test, and magnetic resonance imaging (MRI) study. Gaze nystagmus was observed in four patients (80%), while abnormal pursuit, saccade, and optokinetic nystagmus tests were noted in all patients (100%). MRI demonstrated infarction at the ponto-medullary junction in one patient and upper medulla in one patient. Both patients revealed caloric areflexia and normal VEMPs. In contrast, another three patients with infarction at the middle inferior olive level, all displayed abnormal (including absent or delayed) VEMPs, and one patient showed caloric areflexia. Topographical correlations of lateral medullary infarction with caloric and VEMP tests reveal that caloric areflexia is possibly linked with rostrally located infarction, while absent or delayed VEMPs relate to caudally located infarction. PMID- 19562360 TI - Robotic surgery in ear nose and throat. AB - The arrival of a commercial surgical robotic platform at our institution has raised the question of its application and usefulness within the department of otolaryngology head and neck surgery. In order to answer this question, we sought to perform a qualitative review to examine the evolution of commercial surgical robotics and examine present and future applications of this emerging technology within our specialty. The main objective of this study is to examine the development and application of robotic surgery in otolaryngology, head and neck surgery. The study includes a qualitative systematic review. We have reviewed research papers and studies that specifically relate to the use of robots in otorhinolaryngology. More specifically, we have attempted to review those studies that have significantly added to the development of this field. In summary, we have examined eight animal studies, six cadaveric studies, nine human trials. Robotic surgery in ENT is a safe and feasible option. In certain procedures, it offers significant benefits over conventional surgery. Instrument and robotic arm size, and costs are limiting factors that prevent the use of robots being applied to many additional ENT procedures. We feel the development of new speciality specific robots will yield a new era in the common use of robotics in ENT. PMID- 19562361 TI - Does dictating the letter to the GP in front of a follow-up patient improve satisfaction with the consult? A randomised controlled trial. AB - Various studies have shown that receiving a copy of the letter to the General Practitioner (GP) improves patient satisfaction with the consult. We aimed to establish whether dictating the letter to the GP in front of a listening patient does likewise. Follow-up patients have shorter allotted consultation times. This may contribute to dissatisfaction, hence the choice of our target group. One hundred consecutive follow-up patients who met the eligibility criteria were randomised to listen to the GP letter or not. Immediately after the consult, they were asked to fill in a questionnaire which, in addition to enquiring about various aspects of the consult asked them to quantify their overall satisfaction by means of a ten-point graded visual analogue score. Forty-nine patients received dictation. The mean age and sex distribution of the two groups were matched. The median overall satisfaction in the dictation and non-dictation groups were ten and eight, respectively, this was statistically significant. There was no significant difference between patients' rating of whether the consult had addressed their ailment adequately, explanation(s) given or the length of consult. Sixty-one percent of patients in the non-dictation group would like to have listened to the dictation, whilst all patients in the dictation group found it useful. This study is the first of its kind in the ENT population. Dictating a letter to the GP in front of a listening patient led to a statistically significant improvement in satisfaction independent of possible confounding aspects of the consult. PMID- 19562362 TI - The effect of a Heat and Moisture Exchanger (Provox HME) on pulmonary protection after total laryngectomy: a randomized controlled study. AB - The goal of this randomized controlled study was to investigate the effect of Heat and Moisture Exchanger use on pulmonary symptoms and quality of life aspects in laryngectomized patients. Eighty laryngectomized patients were included and randomized into an HME and Control group. The effect of the HME was evaluated by means of Tally Sheets and Structured Questionnaires. The results showed a significant decrease in the frequency of coughing, forced expectoration, and stoma cleaning in the HME group. There were trends for the prosthetic speakers to report more fluent speech with the HME and for the HME group to report fewer sleeping problems. In conclusion, this study, performed in Poland, confirms the results of previous studies performed in other countries, showing that pulmonary symptoms decrease significantly with HME use and that related aspects such as speech and sleeping tend to improve, regardless of country or climate. PMID- 19562363 TI - Magnet displacement: a rare complication following cochlear implantation. AB - The purpose of this paper is to describe cases which reported complication after cochlear implantation in children: displacement of magnet from the receiver pocket, possibly aided by the use of magnetic toys. We observed magnet displacement in two female children from the same family and in one male child. Age at implantation was 23, 51, and 24 months, respectively. Magnet displacement occurred at 37, 16, and 32 months, respectively after the initial surgery. The magnets were replaced under general anaesthesia and we did not observe recurrent magnet dislodgement. Measurements indicated that forces required to remove the magnet from its pocket were not greater than those exerted by magnetic toys or the magnet used in the external sender coil. Although magnet displacement is not common after cochlear implantation, it is a major complication in children where subsequent general anaesthesia and surgery are necessary to replace the magnet. Therefore, we propose that pockets for removable magnets of cochlear implants used in children should be redesigned to increase forces to remove the magnet or that removable magnets not be used at all. PMID- 19562364 TI - Lipomas of the oral cavity: clinical and histopathologic study of 41 cases in a Brazilian population. AB - This study evaluated the clinical and histopathologic aspects of 41 cases of oral lipomas diagnosed in a Brazilian population. All records from patients diagnosed with oral lipoma between 1970 and 2008 were reviewed. Histological sections were evaluated by light microscopy. There was a predominance of females (2.4:1), with a peak incidence between the sixth and seventh decade. The buccal mucosa was the most affected site (53.7%), followed by the buccal sulcus (14.6%) and tongue (9.8%). Tumor size ranged from 0.5 to 10 cm and the mean reported duration was 48 months. Histologically, the following variants were identified: lipoma (41.5%), fibrolipoma (34.1%), spindle cell lipoma (9.8%), sialolipoma (9.8%), osteolipoma (2.4%), and chondrolipoma (2.4%). Most tumors were well delimited, irrespective of the variant. Lipomas are rare tumors of the oral cavity. The characterization of new variants, such as sialolipomas, and the identification of histological subtypes in already known variants, such as low-fat and fat-free spindle cell lipomas, highlight the importance for careful microscopic evaluation of these tumors, which might be combined with immunohistochemistry in some cases. PMID- 19562365 TI - Intraoperative real-time (99m)Tc-sestamibi scintigraphy with miniature gamma camera allows minimally invasive parathyroidectomy without ioPTH determination in primary hyperparathyroidism. AB - BACKGROUND: Minimally invasive parathyroidectomy (MIP) is being widely accepted as the procedure of choice for the treatment of primary hyperparathyroidism (PHPT), which is caused by a parathyroid single adenoma in more than 80% of cases in some series. Preoperative location studies, like sestamibi scans, allow the proper identification of pathologic gland and intraoperative parathormone (ioPTH) assay is used to confirm the removal of the adenoma. We have studied the feasibility of a new miniature gamma camera (MGC) used intraoperatively to locate parathyroid adenomas and confirm its correct excision. MATERIALS AND METHODS: Twenty patients with PHPT positively diagnosed by preoperative sestamibi scans underwent a MIP. In the first five patients, both ioPTH assay and the new hand held MGC were used consecutively to locate and confirm the excision of the pathologic gland. For the next 15 cases, PTH was measured but not used intraoperatively for diagnosis and the MGC was the only diagnostic tool employed to perform the operation. Concordance between preoperative and intraoperative scintigraphy, surgical time, success rate, and complications are analyzed. RESULTS: All cases were operated on successfully by a MIP. After 1 year follow up, the drop of PTH and the normalization of calcium levels confirmed the excision of all pathologic tissue. The MGC proved its usefulness in all patients offering accurate real-time intraoperative images for location and confirming the success of the procedure. CONCLUSIONS: The MGC is a useful instrument in MIP for PHPT. It may be used as complementary to the standard tools used to date, or even replace them, at least in selected cases of single adenomas. PMID- 19562366 TI - The glycocalyx maintains a cell surface pH nanoenvironment crucial for integrin mediated migration of human melanoma cells. AB - The glycocalyx consists of proteoglycans, glycoproteins, glycosaminoglycans, associated plasma proteins, and soluble glycosaminoglycans and covers the surface of all eukaryotic cells. It mediates specific recognition events, modulates biological processes such as ligand-receptor interactions, and has been proposed to affect tumor metastasis. Here, we studied whether the glycocalyx is required for melanoma cell migration. We diminished the glycocalyx of human melanoma cells by inhibiting posttranslational N-glycosylation or by enzymatic digestion of the N-glycosides. This partial destruction of the glycocalyx reduced melanoma cell migration by up to 60%. It was accompanied by the disintegration of a characteristic pH nanoenvironment typically surrounding migrating cells. Restoring this pH profile by stimulating the activity of the Na(+)/H(+) exchanger NHE1 rescued cell migration even in the absence of an intact glycocalyx. The effects of partially removing the glycocalyx compared to those of knocking down beta(1)-integrin expression points to a close functional correlation between glycocalyx, integrins, and cell surface pH nanoenvironment. We conclude that the glycocalyx is required for tumor cell migration. It stabilizes the cell surface pH nanoenvironment allowing a concerted pH-dependent interaction of adhesion receptors and extracellular matrix. PMID- 19562367 TI - Arginine transport in human erythroid cells: discrimination of CAT1 and 4F2hc/y+LAT2 roles. AB - Since arginine metabolites, such as nitric oxide and polyamines, influence the expression of genes involved in erythroid differentiation, the transport of the cationic amino acid may play an important role in erythroid cells. However, available data only concern the presence in these cells of CAT1 transporter (system y(+)), while no information exists on the role of the heterodimeric transporters of system y(+)L (4F2hc/y(+)LAT1 and 4F2hc/y(+)LAT2) which operates transmembrane arginine fluxes cis-inhibited by neutral amino acids in the presence of sodium. Using erythroleukemia K562 cells and normal erythroid precursors, we demonstrate here that arginine transport in human erythroid cells is due to the additive contributions of a leucine-sensitive and leucine insensitive component. In both cell types, leucine inhibition of arginine influx is much less evident in the absence of sodium, a hallmark of system y(+)L. In K562 cells, N-ethylmaleimide, a known inhibitor of CAT transporters (system y(+)), suppresses only a fraction of arginine influx corresponding to leucine insensitive uptake. Moreover, in Xenopus oocytes coexpressing 4F2hc and y(+)LAT2, leucine exerts a marked inhibition of arginine transport, partially dependent on sodium, while no inhibition is seen in oocytes expressing CAT1. Lastly, silencing of SLC7A6, the gene for y(+)LAT2, lowers arginine transport and doubles the intracellular content of the cationic amino acid in K562 cells. We conclude that arginine transport in human erythroid cells is due to both system y(+) (CAT1 transporter) and system y(+)L (4F2hc/y(+)LAT2 isoform), which mainly contribute, respectively, to the influx and to the efflux of the cationic amino acid. PMID- 19562368 TI - RT patterns and chunks in SRT tasks: a reply to Jimenez (2008). AB - Previous research has shown that the formation of units or chunks contributes to sequence learning in serial reaction time (SRT) tasks (Koch & Hoffmann, Psychological Research 63:22-35, 2000). However, some of these results were assumed to be unrelated to sequence learning and to reflect preexistent response tendencies (Jimenez, Psychological Research 72:387-396, 2008). In the Experiment of this study, we aimed to evaluate this issue. One group of participants responded to a strongly structured sequence of digits by pressing one out of six response keys depending on digit identity. In a second experimental group, a weakly structured sequence was presented, which contained comparable transitions among the single items, but did not have series of ascending and descending triplets of successive digits. The results indicated that serial learning in general as well as response tendencies to certain fragments of the sequence were modulated by the manipulation of the strength of relational patterns. The data are consistent with the notion that relational patterns contribute to the formation of chunks as suggested in the original study. PMID- 19562369 TI - Pulmonary histiocytic sarcoma mimicking pulmonary Langerhans cell histiocytosis in a young adult presenting with spontaneous pneumothorax: a potential diagnostic pitfall. AB - We present a case of a histiocytic sarcoma incidentally detected in peripheral lung tissue resected for a spontaneous pneumothorax. Furthermore, we discuss the practical approach to pulmonary Langerhans cell histiocytosis, the main differential diagnosis of this lesion in the lung, based on morphological and immunohistochemical features. A 23-year-old male patient presented with recurrent pneumothoraces. The pulmonary tissue showed a single round granuloma-like lesion measuring 4 mm in diameter in close neighbourhood to a bronchial wall. The granuloma consisted of histiocytic cells with enlarged pale nuclei, plasma cells, lymphocytes and scanty eosinophilic granulocytes giving the impression of a granuloma of pulmonary Langerhans cell histiocytosis on haematoxylin and eosin (H&E) stains. Immunohistochemically, the histiocytic cells were negative for CD1a and S-100. They were positive for CD68, HLA-DR, CD14, CD4, CD11c, CD45LCA and lysozyme. MIB1 (Ki67) showed a nuclear staining of approximately 10% of the histiocytic cells. In summary, these findings were in keeping with a histiocytic sarcoma, a rare haematopoetic neoplasm. By demonstrating this particular case, we emphasise the importance of proving the diagnosis of pulmonary Langerhans cell histiocytosis by means of immunohistochemistry. In case of a negative CD1a reaction in a histiocytic lesion, further immunohistochemical studies have to be performed in order not to misdiagnose a malignant haematopoetic lesion. PMID- 19562371 TI - Determinants of obesity in the Ulm Research on Metabolism, Exercise and Lifestyle in Children (URMEL-ICE). AB - We investigated the prevalence of overweight and obesity in German schoolchildren and analyzed determinants of overweight. In the context of a randomized intervention study, a baseline cross-sectional assessment was carried out in 2006. During a physical examination, height, weight, skin fold thickness, and upper arm and waist circumferences were measured according to a standardized protocol among 1.079 children aged 6-9 years. Overweight and obesity were classified according to the definitions of the International Obesity Task Force. Parents completed a questionnaire on potential determinants of overweight. Logistic regression models were calculated for determinants of overweight and obesity. The prevalence of overweight was 16.5% in boys and 17.3% in girls and of obesity 3.5% and 3.6%, respectively. Migration (29.4 %) was correlated with overweight and obesity. In particular, among boys with migration background, overweight (24.0%) and obesity (6.6%) were highly prevalent. Higher obesity prevalence was associated with maternal smoking during pregnancy, parental overweight, and low parental education. Indicators for physical inactivity such as watching television more than 1 h per weekday, participation in club sports less than once a week, consumption of sweetened drinks (>or=3 times per week), and skipping breakfast before school were associated with childhood obesity. Our results provide further evidence that parental factors such as migration background and education are strongly associated with body mass of the offspring. Physically inactive children with regular consumption of sweetened drinks and no breakfast were prone to be overweight or obese. Changes of these lifestyle factors as targets of interventions are promising to prevent childhood obesity. PMID- 19562372 TI - Maternal bariatric surgery: adverse outcomes in neonates. AB - BACKGROUND: The obesity epidemic in developed countries has led to an increased prevalence of obese women of reproductive age. As maternal obesity has far reaching consequences for both mother and child, the consensus is that weight loss before pregnancy will reduce obesity-related morbidity and mortality. Therefore, an increasing number of women become pregnant after undergoing obesity surgery. RESULTS AND DISCUSSION: From the literature, data shows that perinatal outcome after bariatric surgery is generally considered as favourable for both mother and child. Only a few case reports highlight the possibility of side effects on the foetus and neonate. We report on five cases with severe intracranial bleeding, all possibly related to vitamin K deficiency following maternal bariatric surgery. CONCLUSION: These reports indicate that careful nutritional follow-up during pregnancy after obesity surgery is mandatory, because nutritional deficiencies such as vitamin K deficiency can lead to life threatening bleeding. PMID- 19562373 TI - Relative toxicity of neem fruit, bitter gourd, and castor seed extracts against the larvae of filaria vector, Culex quinquefasciatus (Say). AB - In search of a natural larvicide, petroleum ether, carbon tetrachloride, and methanol extracts of Azadirachta indica fruits and seed extracts of bitter gourd (Momordica charantia) and castor (Ricinus communis) were tested for larvicidal activity against Culex quinquefasciatus. Among the extracts tested, the methanol extract of Az. indica was observed the most potent with LC(50) at 74.04 and 58.52 ppm and LC(-90) at 201.83 and 171.70 ppm as compared to methanol extract of M. charantia with LC(50) at 101.18 and 93.58 ppm and LC(90) at 322.81 and 302.62 ppm carbon tetrachloride extract of R. communis with LC(50) at 144.11 and 92.44 ppm and LC(90) at 432.42 and 352.89 ppm after 24 and 48 h, respectively. The methanol extract of Az. indica exhibited potential results and can be exploited as a preferred natural larvicide for the control of filarial vector, Cx. quinquefasciatus. PMID- 19562370 TI - Molecular genetic analysis of podocyte genes in focal segmental glomerulosclerosis--a review. AB - This review deals with podocyte proteins that play a significant role in the structure and function of the glomerular filter. Genetic linkage studies has identified several genes involved in the development of nephrotic syndrome and contributed to the understanding of the pathophysiology of glomerular proteinuria and/or focal segmental glomerulosclerosis. Here, we describe already well characterized genetic diseases due to mutations in nephrin, podocin, CD2AP, alpha actinin-4, WT1, and laminin beta2 chain, as well as more recently identified genetic abnormalities in TRPC6, phospholipase C epsilon, and the proteins encoded by the mitochondrial genome. In addition, the role of the proteins which have shown to be important for the structure and functions by gene knockout studies in mice, are also discussed. Furthermore, some rare syndromes with glomerular involvement, in which molecular defects have been recently identified, are briefly described. In summary, this review updates the current knowledge of genetic causes of congenital and childhood nephrotic syndrome and provides new insights into mechanisms of glomerular dysfunction. PMID- 19562374 TI - In vitro and in vivo efficacies of ivermectin and cypermethrin against the cattle tick Hyalomma anatolicum anatolicum (Acari: Ixodidae). AB - This study investigated the comparative efficacy of ivermectin and cypermethrin pour-on, for the treatment of Hyalomma anatolicum (a.) anatolicum infestations in bovines. For examining acaricidal efficacy, 480 ticks were exposed in vitro to graded doses of both the acaricides and in vivo efficacy was examined in 360 tick infested bovines treated at the recommended doses of ivermectin (IVM) and cypermethrin (CYM) pour-on. The comparative quantitative assessment of tick burden was done on days 0, 5, 10, 15, and 20 after treatment using "finger counting." The results of the tick survival assay indicated both compounds were effective in vitro against H. a. anatolicum. The arc transformed mean surviving ticks, 24 h post immersion, was 2.66 and zero in groups treated with the highest dilutions of IVM and CYM, respectively. At 15 days post-treatment, the CYM pour on showed a higher in vivo efficacy (no surviving ticks) compared to IVM (mean of 20 surviving ticks). A single dose of CYM and IVM was found effective for 20 and 15 days post-treatment, respectively. Additionally, a questionnaire was used to gather information from 30 small holder dairy farms on the farmer's approach toward the control of ticks. The majority (90%) of respondents were using acaricides incorrectly along with poor husbandry practices on their farms. Overuse of IVM in the tested area of Pakistan may be the reason the IVM is not as effective as expected. These results provide useful tools for the decision making in tick control, as well as providing the basis for testing the findings on provincial and national levels in future studies. PMID- 19562375 TI - Anti-juvenile activity of Azadirachta indica extract on the development and morphometry of filaria vector, Culex quinquefasciatus (Diptera: Culicidae) Say. AB - Efficacy of extracts of Ageratum conyzoides, Argemone mexicana, and Azadirachta indica were tested against Culex quinquefasciatus larvae. Petroleum ether extracts of A. conyzoides and A. mexicana with LC(50) values 425.60 and 267.90 ppm and 140.15 and 137.70 ppm after a period of 24 and 48 h, respectively, were found effective. However, the methanol extract of A. indica with LC(50) values 21.95 and 11.30 ppm after 24 and 48 h of exposure, respectively, was the most competent larvicide. The methanol extract of A. indica, being the most potent extract was therefore studied for its impact on the developmental scenario of the mosquito. It had a significant effect on the life cycle of the vector, C. quinquefasciatus. Hatching was delayed and its rate was reduced compared to control. Larval and pupal development was arrested resulting in decreased pupal transformation and adult emergence. Larval and pupal periods were prolonged with appearance of larval-pupal and pupal-adult intermediates, with an overall increase in the developmental period. Disrupted egg shells, dechitinized body walls, and distorted intestines were observed, indicating clearly the anti juvenile potential of the extract. The growth index was considerably reduced. PMID- 19562376 TI - In vitro antifilarial activity of glutathione S-transferase inhibitors. AB - Female adult bovine filarial worms Setaria digitata were extracted with phosphate buffered saline (pH 7.4) and glutathione S-transferase (GST) activity and protein content were determined. The protein content, GST enzyme activity, and specific activity were 10.61 +/- 3.41 mg ml(-1), 0.09 +/- 0.019 micromol min(-1) ml(-1), and 0.009 +/- 0.002 micromol min(-1) mg(-1) protein, respectively. The GST inhibition studies were performed with and without the inhibitors resulted from earlier molecular docking studies viz., ethacrynic acid, plumbagin, and curcumin for which the IC(50) values were 19.42, 51.41, and 114.86 microM, respectively. The in vitro macrofilaricidal activity of these molecules was studied by worm motility and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay at 24- and 48-h incubation. Plumbagin and ethacrynic acid showed 100% inhibition in worm motility at lower concentrations of 3.19 and 6.6 microM, respectively, at 48-h incubation while curcumin was effective at 54.29 microM. In MTT reduction assay, the ED(50) values (50% inhibition in formazan formation) for plumbagin, ethacrynic acid, and curcumin at 48-h incubation were 1.20, 2.48, and 19.86 microM, respectively. MTT reduction assay showed that plumbagin was the most effective in killing the adult S. digitata worms followed by ethacrynic acid and curcumin. In conclusion, all the three molecules selected by molecular modeling and docking studies inhibited the GST enzyme isolated from S. digitata and exhibited macrofilaricidal activity in vitro. PMID- 19562377 TI - In vitro effects of resveratrol on Trichinella spiralis. AB - Resveratrol, a natural phytoalexin found mainly in grapes, possesses antibacterial, antifungal, and antiviral properties. However, there is no information about its effects on helminths such as Trichinella sp. In the present study, we investigated the effects of resveratrol on the viability of Trichinella spiralis life stages in vitro. Adult forms, newborn larvae (NBL), and muscle larvae (ML) were incubated with resveratrol at concentrations varying from 12.5 to 200 microg/ml. Resveratrol showed significant anthelmintic activity against NBL and adult forms of Trichinella, but not against ML. Our results suggest that resveratrol may be useful as a therapeutic agent to treat trichinellosis in early stages and warrant its further assessment in animal models of disease. PMID- 19562378 TI - Genetic evidence for conspecificity between Dermacentor marginatus and Dermacentor niveus. AB - Ticks are hematophagous arthropods transmitting several harmful human and animal pathogens like viruses, Rickettsia, bacteria, and protozoa. The identification and speciation of ticks were normally performed in Iran using identification key of Arthur (1960) and Kaiser and Hoogstraal (J Parasitol 49:130-139, 1963) or on the basis of morphological characteristic keys recommended by Walker et al. (2003). Although these identification keys are well prepared, but there are in some cases due to the strong overlapping characteristics between species like Dermacentor marginatus and Dermacentor niveus accompanied with serious problems. D. marginatus and D. niveus have been intermittently used synonymously and there is no a generally agreement with the specification of these species. To find out more about these two species, we have analyzed the complete nucleotide sequence of ITS-2 region. Interestingly, we found indeed a sequence homology of 99% between nucleotide sequence of ITS-2 region of D. marginatus and D. niveus. Since the nucleotide sequence of ITS-2 region of D. marginatus in Iran has 98% sequence homology to the other in GenBank registered ITS-2 sequence of D. marginatus, and the morphological characteristics between both examined species showed minimal differences, therefore we believe that the D. marginatus and D. niveus could belong to the same species and 1% differences in nucleotide sequence of ITS-2 region between these two species can be understand as an intra-species polymorphism. PMID- 19562379 TI - 18S ribosomal DNA genotypes of Acanthamoeba species isolated from contact lens cases in the Philippines. AB - This study was carried out to document the genotypes of Acanthamoeba present in contact lens cases from 50 randomly selected contact lens wearers living in Quezon City, Metro Manila, Philippines. Acanthamoeba species were isolated from eight (16%) in 50 contact lens cases examined. We analyzed partial 18S ribosomal DNA (Rns) sequences of the eight isolates and found that the sequence differences were sufficient to distinguish the genotypes. After the isolates were genotyped, using the Basic Local Alignment Search Tool program, their phylogenetic positions relative to known Acanthamoeba isolates were determined. The model-based (GTR+Gamma+Iota) neighbor-joining, maximum likelihood, and Bayesian inference analyses, as well as the non-model-based maximum parsimony analysis were used. Results showed that of the eight isolates, six were Rns genotype T5 while two were Rns genotype T4. This present study indicates that genotype T5 is also a common contaminant in contact lens storage cases. PMID- 19562380 TI - Acaricidal efficacy of thymol on engorged nymphs and females of Rhipicephalus sanguineus (Latreille, 1808) (Acari: Ixodidae). AB - The present work was aimed at evaluating the "in vitro" efficacy of different concentrations of thymol on engorged nymphs and females of Rhipicephalus sanguineus. The nymphs were separated in seven groups and immersed in different concentrations of thymol (0.25%, 0.5%, 1.0%, 1.5%, and 2.0%) for 5 min. A control group was established (water + dimethylsulfoxide) together with a positive control group (Amitraz*), and mortality was evaluated after 15 days. In order to perform the experiment with females, the same methodology was used, and the following concentrations were tested: 1.0%, 1.5% and 2.0%. After 5 min of immersion, the engorged females were fixed on Petri dishes with the aid of a double-sided adhesive tape and kept in a climatized room regulated at 27 +/- 1 degrees C and UR >80 +/- 10%, and different parameters referring to the biology of the non-parasitary phase were evaluated daily. The values found for thymol efficacy on nymphs were 0.0%, 100%, 100%, 100%, and 100% in concentrations of 0.25%, 0.5%, 1.0%, 1.5%, and 2.0%, respectively. In the experiment with engorged females, thymol did not induce any significant alterations (p < 0.05) in the parameters of weight alteration, egg mass weight, pre-oviposition period, hatching percentage, egg production index and nutritional index; however, it affected the engorged females final weight in all treatments (p > 0.05). The concentration of thymol 2% was the one that showed a better efficacy (41%). It was concluded that thymol had a more accentuated deleterious effect on engorged nymphs, and it might be a promising supporting agent for the control of this ixodid. PMID- 19562381 TI - Variably male-biased sex ratio in a marine bird with females larger than males. PMID- 19562383 TI - Modelling the wind damage probability in forests in Southwestern Germany for the 1999 winter storm 'Lothar'. AB - The wind damage probability (P (DAM)) in the forests in the federal state of Baden-Wuerttemberg (Southwestern Germany) was calculated using weights of evidence (WofE) methodology and a logistic regression model (LRM) after the winter storm 'Lothar' in December 1999. A geographic information system (GIS) was used for the area-wide spatial prediction and mapping of P (DAM). The combination of the six evidential themes forest type, soil type, geology, soil moisture, soil acidification, and the 'Lothar' maximum gust field predicted wind damage best and was used to map P (DAM) in a 50 x 50 m resolution grid. GIS software was utilised to produce probability maps, which allowed the identification of areas of low, moderate, and high P (DAM) across the study area. The highest P (DAM) values were calculated for coniferous forest growing on acidic, fresh to moist soils on bunter sandstone formations-provided that 'Lothar' maximum gust speed exceeded 35 m s(-1) in the areas in question. One of the most significant benefits associated with the results of this study is that, for the first time, there is a GIS-based area-wide quantification of P (DAM) in the forests in Southwestern Germany. In combination with the experience and expert knowledge of local foresters, the probability maps produced can be used as an important tool for decision support with respect to future silvicultural activities aimed at reducing wind damage. One limitation of the P (DAM)-predictions is that they are based on only one major storm event. At the moment it is not possible to relate storm event intensity to the amount of wind damage in forests due to the lack of comprehensive long-term tree and stand damage data across the study area. PMID- 19562382 TI - [Impact of physiotherapy, massages and lymphatic drainage in migraine therapy]. AB - Physical treatment, spinal manipulative therapy and massages are often recommended to treat migraine as a prophylactic therapy. Clinical experience and theoretical concepts support their usefulness. However, data on these therapies are scarce. On the basis of the available studies, it is impossible to determine whether or not these therapies are effective. There is a lack of well-designed prospective, randomized controlled trials with a sufficiently long follow-up to observe these therapies. Due to the high acceptance of physical treatment on the one hand and preconceptions about drug treatment on the other, these types of therapies may be an alternative option for some patients if their efficacy is established. A cost-benefit analysis of theses therapies should consider the long amount of time required for them compared with drug intake. PMID- 19562384 TI - Prevalence and influence of malnutrition on quality of life and performance status in patients with locally advanced head and neck cancer before treatment. AB - GOALS OF WORK: The aim of this study was to evaluate the prevalence and influence of malnutrition (unintentional weight loss >or=5% in the last 3 months) on quality of life (QoL) and performance status (PS) in head and neck cancer patients (HNC) before treatment. PATIENTS AND METHODS: Sixty-one consecutive outpatients affected by locally advanced HNC (III-IVA stage) were enrolled. In all patients, nutritional intake (by diet history), nutritional status (Patient Generated Subjective Global Assessment), unintentional weight loss (UWL), serum prealbumin, hemoglobin level (Hb), C-reactive protein, QoL (European Organization for Research and Treatment of Cancer Quality of Life Questionnaire C-30 v. 3.0), and PS (Eastern Cooperative Oncology Group (ECOG) PS) were assessed before radio or concomitant chemoradiotherapy. MAIN RESULTS: Thirty-six percent of HNC were malnourished before treatment. The median ECOG PS in malnourished patients was 1 (0-2), whereas in nonmalnourished was 0 (0-2; p = 0.018). Physical (p = 0.043), role (p = 0.047), and social functions (p = 0.024) scores were significantly worse in malnourished than in nonmalnourished HNC. Fatigue (p < 0.001), appetite loss (p < 0.001), and nausea and vomiting (p = 0.002) scores were worse in malnourished patients than in nonmalnourished. In the multivariate analysis, UWL and Hb level independently influenced physical (p = 0.002; p = 0.005), role (p = 0.004; p = 0.001), and social functions (p = 0.024; p = 0.009). CONCLUSION: Our data suggest that an early and intensive nutritional support might reduce weight loss before, during, and after treatment completion, improving outcome, QoL, and PS. PMID- 19562385 TI - Postural control in patients with lumbar disc herniation in the early postoperative period. AB - Chronic spinal disc disease leads to disorders in postural movement coordination. An incorrect asymmetrical movement pattern for the lower limbs loading impairs proprioception and deteriorates postural stability, particularly when the vision is occluded. The standard surgical treatment improves biomechanical conditions in the lumbar spine, reduces pain, yet does it reduce the stability deficit in the upright position? An answer to the latter question would help work out targeted therapy to improve postural stability. We hypothesized that the standard surgical treatment would improve postural stability reflected by decreased sway variability accounting for better use of proprioceptive inputs postoperatively. Thirty-nine patients with lumbar disc herniation participated in the study. Their postural sway was recorded in anterior/posterior and medial/lateral planes with their eyes open or closed (EC) before and after surgery. The variability, range, mean velocity of the recorded time series and the area of the ellipse enclosed by the statokinesiogram were used as measures of postural stability. Preoperatively, EC condition resulted in an increased variability and mean velocity of postural sway, while postoperatively it caused an increase in sway mean velocity and sway area only with no effect on sway variability and range. The comparison of the balance before and after the surgery in the EC condition showed significant decrease in all parameters. In the early postoperative period, the patients recover the ability to control their postural sway in EC within normal limits, however, at the expense of significantly increased frequency of corrective torques. It is probably a transient short-term strategy needed to compensate for the recovery phase when the normal weighting factors for all afferents are being reestablished. We propose that this transient postoperative period may be the best timing of therapeutic intervention targeted at facilitating and reinforcing the acquisition of correct motor patterns. PMID- 19562386 TI - Interspinous implants (X Stop, Wallis, Diam) for the treatment of LSS: is there a correlation between radiological parameters and clinical outcome? AB - Neurogenic intermittent claudication, caused by lumbar spinal stenosis (LSS), usually occurs after the age of 50 and is one of the most common degenerative spinal diseases in the elderly. Among patients over the age of 65 with LSS, open decompression is the most frequently performed spinal operation. The recently introduced interspinous spacers are a new alternative under discussion. In this retrospective study, we reviewed medical records and radiographs of patients with LSS and NIC treated from June 2003 to June 2007. All included patients (n = 129) were treated with interspinous implants (X Stop Wallis, or Diam). Evaluations of pain, using a visual analog scale (VAS), and radiographic signs, using two-plane X-rays of the lumbar spine, were performed preoperatively (preop), postoperatively (postop) and after discharge (FU 2-3). Gender ratio (m:w) was 1.1:1. Mean age of the patients was 60.8 +/- 16.3 years. Foraminal height, foraminal width, foraminal cross-sectional area, intervertebral angle, as well as anterior and posterior disc height changed significantly (P < 0.0001) after implantation of the interspinous device. Postoperatively, symptom relief (VAS) was significant (P < 0.0001). The X Stop implant improved (in some cases significantly) the radiographic parameters of foraminal height, width, and cross sectional area, more than the Diam and Wallis implants; however, there was no significant difference among the three regarding symptom relief. FU 1 was on average 202.3 +/- 231.9 and FU 2 527.2 +/- 377.0 days postoperatively. During FU, the radiological improvements seemed to revert toward initial values. Pain (VAS) did not increase despite this "loss of correction." There was no correlation between age and symptom improvement. There was only very weak correlation between the magnitude of radiographic improvement and the extent of pain relief (VAS). The interspinous implant did not worsen low-grade spondylolisthesis. Provided there is a strict indication and fusion is not required, implantation of an interspinous spacer is a good alternative to treat LSS. The interspinous implant offers significant, longlasting symptom control, even if initially significant radiological changes seem to revert toward the initial values ("loss of correction"). PMID- 19562387 TI - Effects of gender and cognitive-behavioral management of depressive symptoms on rehabilitation outcome among inpatient orthopedic patients with chronic low back pain: a 1 year longitudinal study. AB - Psychological factors have been found to be of major importance for the transition from acute to chronic low back pain (CLBP). Although some evidence has been provided that depressive symptoms occur secondarily to CLBP, psychological treatment modules that specifically address depressive symptoms are not yet included in German inpatient rehabilitation programs. In this study, a standard rehabilitation program for patients with CLBP and depressive symptoms was compared to a standard rehabilitation, into which a cognitive-behavioral management training of depressive symptoms was integrated. Moreover, treatment effects of this multidisciplinary standard rehabilitation program delivered to patients with either no or only mild depressive symptoms were investigated. As a further aim of the present study, gender effects on rehabilitation outcomes were examined. Short-, mid-, and long-term effects on individual global improvement as well as pain-related, psychological, and work-related measures were evaluated among N = 199 consecutively admitted patients with CLBP, aged from 24 to 62 years. The standard rehabilitation program had no persisting effects on psychological outcomes among patients with no and mild depressive symptoms. Patients with moderate and severe depressive symptoms in the standard rehabilitation program did not benefit with clinical importance at the 6- and 12 month follow-up assessments, but did show clinically significant improvements in psychological outcome measures at the 6-month follow-up assessment when the supplemental psychological component was applied. Additionally, days of sick leave decreased in the intervention group in the mid-term when compared to the baseline. Females benefited more in mental health than males. However, due to regression effects at the 12-month follow-up assessment, booster sessions are highly recommended. The results presented here support the notion that a more adequately tailored rehabilitation program seems to improve rehabilitation success and prevent further development of CLBP among this high-risk subpopulation. PMID- 19562389 TI - How to Tango: a manual for implementing Spine Tango. AB - The generic approach of the Spine Tango documentation system, which uses web based technologies, is a necessity for reaching a maximum number of participants. This, in turn, reduces the potential for customising the Tango according to the individual needs of each user. However, a number of possibilities still exist for tailoring the data collection processes to the user's own hospital workflow. One can choose between a purely paper-based set-up (with in-house scanning, data punching or mailing of forms to the data centre at the University of Bern) and completely paper-free online data entry. Many users work in a hybrid mode with online entry of surgical data and paper-based recording of the patients' perspectives using the Core Outcome Measures Index (COMI) questionnaires. Preoperatively, patients can complete their questionnaires in the outpatient clinic at the time of taking the decision about surgery or simply at the time of hospitalisation. Postoperative administration of patient data can involve questionnaire completion in the outpatient clinic, the handing over the forms at the time of discharge for their mailing back to the hospital later, sending out of questionnaires by post with a stamped addressed envelope for their return or, in exceptional circumstances, conducting telephone interviews. Eurospine encourages documentation of patient-based information before the hospitalisation period and surgeon-based information both before and during hospitalisation; both patient and surgeon data should be acquired for at least one follow-up, at a minimum of three to six months after surgery. In addition, all complications that occur after discharge, and their consequences should be recorded. PMID- 19562390 TI - An analysis of hydrophobic interactions of thymidylate synthase with methotrexate: free energy calculations involving mutant and native structures bound to methotrexate. AB - Since the human body for many reasons can adapt and become resistant to drugs, it is important to develop and validate computer aided drug design (CADD) methods that could help predict binding affinity changes that can result from these resistant enzymes. The free energy perturbation (FEP) methodology is the most accurate means of estimating relative binding affinities between inhibitors and protein variants. In this paper, we describe the role played by hydrophobic residues lining the active site region, particularly (79)Ile and (176)Phe, in the binding of methotrexate to the Escherichia coli (E. coli) thymidylate synthase (TS) enzyme, using the thermodynamic cycle perturbation (TCP) approach. The computed binding free energy differences on the binding of methotrexate to the native and some mutant E. coli TS structures have been compared with experimental results. Computationally, four different 'mutations' have been simulated on the TS enzyme with methotrexate (MTX): (79)Ile --> (79)Val; (79)Ile --> (79)Ala; (79)Ile --> (79)Leu; and (176)Phe --> (176)Ile. The calculated results indicate that in each of these cases, the native residues ((79) Ile and (176) Phe) interact more favorably with methotrexate than the mutant residues and these results are corroborated by experimental measurements. Binding preference to wild type residues can be rationalized in terms of their better hydrophobic contacts with the phenyl ring of methotrexate. PMID- 19562391 TI - Comparison of two PCR-based methods and conventional culture for the detection of nasal carriage of Staphylococcus aureus in pre-operative patients. AB - Nasal carriage is an important risk factor for the development of post-operative infections with Staphylococcus aureus and pre-operative treatment with mupirocin in carriers reduces the post-operative infection rate. Therefore, it is important to identify nasal carriage rapidly. Two polymerase chain reaction (PCR) techniques were compared to conventional culture in surgical patients. In 404 consecutive patients, nasal swabs were taken for pre-operative screening for the nasal carriage of S. aureus. The performance of the Roche Staphylococcus Kit on Lightcycler (Roche; RSA) and the Becton Dickinson (San Diego, CA) GeneOhm StaphSR assay on Smartcycler (Cepheid; BDSA) were compared with semi-quantitative culture. The sensitivity for culture, RSA and BDSA compared to the gold standard was 98.2, 82.0 and 85.6%, respectively, and the specificity was 100, 98.3 and 99.3%, respectively. The lower sensitivity of both PCR techniques was associated with samples with low bacterial loads. The RSA and BDSA were similar in performance and are suitable for the pre-operative identification of nasal carriers of S. aureus. PMID- 19562388 TI - Prognostic factors in intramedullary astrocytomas: a literature review. AB - Astrocytomas affect a significant portion of patients with intramedullary tumors. These infiltratively growing tumors are treated by a variety of methods -- biopsy and decompressive surgery, maximal safe resection, adjuvant oncological therapy. Also, numerous prognostic factors are reported in the literature. Better understanding of factors that influence prognosis may help in treatment planning with the goal of prolonging survival. We have thus undertaken an extensive literature review in order to define factors affecting prognosis. A total of 38 articles were studied. Only tumor grade was consistently reported as the major factor affecting prognosis. The influence of other clinical factors (age, gender, history length, functional status, tumor location or extent, syrinx or cyst presence) can be speculated upon, but cannot be assessed adequately from the available literature. For both low- and high-grade (HG) astrocytomas, maximal safe tumor resection should be the primary treatment objective but is often not feasible in contrast to other intramedullary and spinal neoplasms. Since the biological nature of spinal cord HG glioma is identical to that of the brain, the same treatment algorithm of maximal safe resection followed by concomitant radio- and chemotherapy would be sensible to implement. PMID- 19562392 TI - Retrospective analysis of the genetic diversity of Klebsiella oxytoca isolated in Poland over a 50-year period. AB - Population genetics analyses and determination of the phylogenetic relationships between strains have proven to be extremely useful approaches, enabling deeper insights into the epidemiological pattern of bacterial species. There is no longitudinal data describing the molecular epidemiology of Klebsiella oxytoca strains that are opportunistic pathogens responsible for an increasing number of multi-resistant infections in hospitals. The aim of the present study was to assess the genetic diversity of K. oxytoca strains over a 50-year period using internal transcribed spacer polymerase chain reaction (ITS-PCR) and PCR MP (ang. PCR melting profiles) genotyping methods on a large collection of strains isolated from the patients of several hospitals in Poland. The phylogenetic analysis based on ITS-PCR exhibited six distinct branches. Two main groups, KoX and KoY, with four and two sub-groups within KoX and KoY, respectively, have been identified. Typing by the PCR MP method showed a higher level of genetic diversity. However, all K. oxytoca strains were also divided into six genotype groups (KoA, KoB, KoC, KoD, KoE and KoF). In conclusion, we found that the ITS PCR and PCR MP methods are useful for the phylogenetic delineation of genetic groups in K. oxytoca. PMID- 19562393 TI - The added value of thorough economic evaluation of telemedicine networks. AB - This paper proposes a thorough framework for the economic evaluation of telemedicine networks. A standard cost analysis methodology was used as the initial base, similar to the evaluation method currently being applied to telemedicine, and to which we suggest adding subsequent stages that enhance the scope and sophistication of the analytical methodology. We completed the methodology with a longitudinal and stakeholder analysis, followed by the calculation of a break-even threshold, a calculation of the economic outcome based on net present value (NPV), an estimate of the social gain through external effects, and an assessment of the probability of social benefits. In order to illustrate the advantages, constraints and limitations of the proposed framework, we tested it in a paediatric cardiology tele-expertise network. The results demonstrate that the project threshold was not reached after the 4 years of the study. Also, the calculation of the project's NPV remained negative. However, the additional analytical steps of the proposed framework allowed us to highlight alternatives that can make this service economically viable. These included: use over an extended period of time, extending the network to other telemedicine specialties, or including it in the services offered by other community hospitals. In sum, the results presented here demonstrate the usefulness of an economic evaluation framework as a way of offering decision makers the tools they need to make comprehensive evaluations of telemedicine networks. PMID- 19562394 TI - Problems with the microbial production of butanol. AB - With the incessant fluctuations in oil prices and increasing stress from environmental pollution, renewed attention is being paid to the microbial production of biofuels from renewable sources. As a gasoline substitute, butanol has advantages over traditional fuel ethanol in terms of energy density and hygroscopicity. A variety of cheap substrates have been successfully applied in the production of biobutanol, highlighting the commercial potential of biobutanol development. In this review, in order to better understand the process of acetone butanol-ethanol production, traditional clostridia fermentation is discussed. Sporulation is probably induced by solvent formation, and the molecular mechanism leading to the initiation of sporulation and solventogenesis is also investigated. Different strategies are employed in the metabolic engineering of clostridia that aim to enhancing solvent production, improve selectivity for butanol production, and increase the tolerance of clostridia to solvents. However, it will be hard to make breakthroughs in the metabolic engineering of clostridia for butanol production without gaining a deeper understanding of the genetic background of clostridia and developing more efficient genetic tools for clostridia. Therefore, increasing attention has been paid to the metabolic engineering of E. coli for butanol production. The importation and expression of a non-clostridial butanol-producing pathway in E. coli is probably the most promising strategy for butanol biosynthesis. Due to the lower butanol titers in the fermentation broth, simultaneous fermentation and product removal techniques have been developed to reduce the cost of butanol recovery. Gas stripping is the best technique for butanol recovery found so far. PMID- 19562395 TI - The tight coupling between category and causal learning. AB - The main goal of the present research was to demonstrate the interaction between category and causal induction in causal model learning. We used a two-phase learning procedure in which learners were presented with learning input referring to two interconnected causal relations forming a causal chain (Experiment 1) or a common-cause model (Experiments 2a, b). One of the three events (i.e., the intermediate event of the chain, or the common cause) was presented as a set of uncategorized exemplars. Although participants were not provided with any feedback about category labels, they tended to induce categories in the first phase that maximized the predictability of their causes or effects. In the second causal learning phase, participants had the choice between transferring the newly learned categories from the first phase at the cost of suboptimal predictions, or they could induce a new set of optimally predictive categories for the second causal relation, but at the cost of proliferating different category schemes for the same set of events. It turned out that in all three experiments learners tended to transfer the categories entailed by the first causal relation to the second causal relation. PMID- 19562396 TI - [Hyperbaric oxygen for carbon monoxide poisoning: case report of severe poisoning due to exhaust fumes]. AB - After rescue and transport a patient with severe carbon monoxide poisoning (CO-Hb concentration 82%) was treated by hyperbaric oxygen (HBO) therapy in a pressure chamber. During decompression the patient suffered cardiac arrest due to massive acidosis, hyperpotassaemia, haemoconcentration and elevated CO-Hb level. After successful resuscitation and cardiopulmonary diagnostics, HBO therapy was continued for prophylactic treatment of delayed neurological sequelae. Three months after completing treatment and discharge from hospital there were no neurological sequelae. PMID- 19562397 TI - [Tension pneumothorax after acute airway displacement due to pulmonary aspergillosis]. AB - Acute lymphoblastic leukaemia is the most common malignancy in childhood. This disease and its associated therapy may lead to specific life-threatening complications if general anaesthesia has to be carried out. The case of a 14-year old boy suffering from aspergillosis because of immunosuppression in the course of chemotherapy is reported. Due to a cerebral round lesion an open biopsy was required. After induction of anaesthesia, severe pulmonary obstruction developed. After exchange of the endotracheal tube a coagulum-like foreign body interspersed with Aspergillus hyphae obstructing the distal aperture in a valve-like manner could be recovered. The resulting unilateral tension pneumothorax had to be relieved with a closed pleural drainage. With reference to this as yet unreported life-threatening complication of pulmonary aspergillosis, the appropriate preparation and conduction of general anaesthesia are discussed. PMID- 19562398 TI - [Importance of abdominal compartment syndrome in Germany: a questionnaire]. AB - BACKGROUND: The abdominal compartment syndrome (ACS) is a life-threatening condition and may affect any critically ill patient. Little is known about the recognition and management of the ACS in Germany. METHODS: A postal questionnaire was sent to departments of surgery and anesthesia of German hospitals with more than 450 beds. RESULTS: From the 222 eligible hospitals a total of 113 replies were received. Most respondents (95%) indicated that the ACS plays a role in their clinical practice. Measurement of intra-abdominal pressure (IAP) is not performed by 26% while it is routinely done by 30%. Intra-abdominal pressure is mostly (94%) assessed via the bladder pressure. Of the respondents 41% measure IAP only in those patients thought likely to develop ACS. Risk factors of the ACS would lead to IAP monitoring in 10-23% of cases. The majority (86%) would require signs of organ dysfunction together with exceeding the IAD threshold in order to opt for a surgical decompression. The attitude towards the critical threshold (>20 mmHg or >25 mmhg) divided respondents into two groups of similar size (39% compared to 47%, respectively). CONCLUSIONS: German anesthesiologists and surgeons are familiar with the ACS. However, about one-quarter never measure IAP and there is a considerable variance as to which patients are at risk to develop ACS and how often IAP should be measured in these patients. This could indicate a lack of acceptance or simply a persisting need for more data concerning the avoidance and treatment of the ACS. PMID- 19562399 TI - [Economic benefits of overlapping induction: investigation using a computer simulation model]. AB - The aim of this study was to investigate the potential economic benefit of overlapping anaesthesia induction given that all patient diagnosis-related groups (AP DRG) are used as the model for hospital reimbursement. A computer simulation model was used for this purpose. Due to the resource-intensive production process, the operating room (OR) environment is the most expensive part of the supply chain for surgical disciplines. The economical benefit of a parallel production process (additional personnel, adaptation of the process) as compared to a conventional serial layout was assessed. A computer-based simulation method was used with commercially available simulation software. Assumptions for revenues were made by reimbursement based on AP DRG. Based on a system analysis a model for the computer simulation was designed on a step-by-step abstraction process. In the model two operating rooms were used for parallel processing and two operating rooms for a serial production process. Six different types of surgical procedures based on historical case durations were investigated. The contribution margin was calculated based on the increased revenues minus the cost for the additional anaesthesia personnel. Over a period of 5 weeks 41 additional surgical cases were operated under the assumption of duration of surgery of 89+/ 4 min (mean+/-SD). The additional contribution margin was CHF 104,588. In the case of longer surgical procedures with 103+/-25 min duration (mean+/-SD), an increase of 36 cases was possible in the same time period and the contribution margin was increased by CHF 384,836. When surgical cases with a mean procedural time of 243+/-55 min were simulated, 15 additional cases were possible. Therefore, the additional contribution margin was CHF 321,278. Although costs increased in this simulation when a serial production process was changed to a parallel system layout due to more personnel, an increase of the contribution margin was possible, especially with procedures of shorter duration (<120 min). For longer surgical times, the additional costs for the workforce result in a reduced contribution margin depending on the models chosen to handle overtime of the technical OR personnel. Important advantages of this approach for simulation are the use of the historical production data and the reflection of the specificities of the local situation. Computer simulation is an ideal tool to support operation room management, particularly regarding the planning of resource allocation and the coordination of workflow. PMID- 19562400 TI - [Mobile learning with podcasts]. PMID- 19562402 TI - [Analgesia as an important component of emergency care]. AB - Besides protection of vital functions, pain treatment is one of the fundamental tasks in emergency care. However, in emergency situations patients frequently do not receive any pain treatment at all or, despite high pain intensities, treatment remains insufficient. There are various reasons for inadequate prehospital analgesia: fear of side effects, underestimation of the duration of emergency care, and concern about impeding diagnostic analysis are the most frequently stated motives. Thereby one should not disregard that pain has several negative effects on the patient that should be avoided. In this article the basics of preclinical analgesia are shown and concepts presented that should help the emergency physician to accomplish adequate analgesia in an emergency situation. PMID- 19562403 TI - Patients' perceptions of health-related quality of life in rheumatoid arthritis. AB - The purpose of this study is to determine how health-related quality of life (HRQL) is perceived by patients with rheumatoid arthritis using textual analysis and to identify associations between sociodemographic and clinical variables and patients' perceptions. Multicentre, cross-sectional study, including 781 patients (78.8% females, mean age 60.65 years, standard deviation 14.22). Sociodemographic and clinical variables were collected. A questionnaire was used which included an open question, "What does HRQL mean to you?" Cluster analysis was used, and answers to the open question were analysed using textual analysis. Cluster analysis showed three typologies differentiated by gender, age, and socioeconomic level and by the terms used to describe perceptions of HRQL with no marked clinical differences. Characteristic phrases were "to be totally independent, to have no pain, and to do things without thinking about the negative effects" (typology 1); "to have no pain and control my moods and my illness has made me a little excitable" (typology 2); and "to feel well enough to do things for myself" (typology 3). Three representative typologies of patients were identified who differed clearly in their perceptions of HRQL. Textual analysis may be considered as a valid tool for the analysis of complex issues such as quality of life. PMID- 19562404 TI - In vitro evaluation of microleakage under orthodontic brackets using two different laser etching, self etching and acid etching methods. AB - This study evaluated the microleakage of brackets bonded by four different enamel etching techniques. Forty freshly extracted human premolars were divided randomly into four equal groups and received the following treatment: group 1, acid etching; group 2, self-etching primer (SEP); group 3, erbium:yttrium-aluminum garnet (Er:YAG) laser etching; and group 4, erbium, chromium:yttrium-scandium gallium-garnet (Er,Cr:YSGG) laser etching. After photopolymerization, the teeth were kept in distilled water for 1 month and then subjected to 500 thermal cycles. Then, the specimens were sealed with nail varnish, stained with 0.5% basic fuchsin for 24 h, sectioned, and examined under a stereomicroscope. In addition, they were scored for marginal microleakage at the adhesive-enamel and bracket-adhesive interfaces from the incisal and gingival margins. Statistical analyses consisted of the Kruskal-Wallis test and the Mann-Whitney U test with Bonferroni correction. Microleakage occurred between the adhesive-enamel and bracket-adhesive interfaces in all groups. For the adhesive-enamel surface, a significant difference was observed between group 1 and groups 2 (P = 0.011), 3 (P = 0.002), and 4 (P = 0.000) on the gingival side. Overall, significant differences were observed between group 1 and groups 3 (P = 0.003) and 4 (P = 0.000). In dental bonding procedures, acid etching was found to result in the least microleakage. Since etching with a laser decreases the risk of caries and is time-saving, it may serve as an alternative to acid etching. PMID- 19562405 TI - Intraoral metal laser welding: a case report. AB - The possibility of laser welding of dental prostheses offers great advantages: first, the operator has the possibility of welding on the master model, which decreases the number of passages and thus the possibility of errors and damage, and secondly, the patient attends only a few sessions, and, due to the possibility of fixing the damaged prostheses, there is no need to resort to the technician's laboratory. In a previous study we described the experimental phases of intraoral welding, from the in vitro model on animal jaws with evaluations of the temperature variations during welding through thermal chamber and type K thermocouples. In this study we describe the intraoral welding in vivo on human subjects by using, as in the previous study, a fibre-delivered neodymium:yttrium aluminum-garnet (Nd:YAG) laser. The in vivo phase allowed a restored prosthesis to be positioned and intraorally welded in the upper central sector with optimal results both in patient's comfort and in aesthetic effects. This first in vivo test confirmed that the use of a laser technique for the intraoral welding of metal prostheses is possible, with no particular problems and risks for the biological structures close to the welding zone. PMID- 19562406 TI - Laparoscopic right hemicolectomy for intestinal intussuception. AB - INTRODUCTION: Adult intussusception is rare and usually associated with carcinoma in 50% of the cases. These have traditionally been managed using an open technique. We herein describe a laparoscopic extended right hemicolectomy in a 62 year-old lady with an intussuception secondary to a transverse colonic tumor. METHODS: The patient presented with 6 weeks of crampy, colicky, abdominal pain. Her CT scan reported intussuception of the proximal large bowel. She underwent an extended laparoscopic right hemicolectomy with primary anastomosis. RESULTS: Her post-operative recovery was uneventful and the histology reported a large bowel adenocarcinoma (pT4, N0, M0) with none out of 25 nodes involved. CONCLUSION: When operative intervention is required, intussuception may be managed using a minimally invasive technique. However, large bowel intussuception in adults may have a malignant cause thus laparoscopic resection should only be performed by surgeons experienced in laparoscopic resections for colorectal malignancies as oncological safety must be the primary concern. This laparoscopic approach facilitates rapid recovery and earlier time to adjuvant therapy if required. PMID- 19562407 TI - Trends in prostate specific antigen testing in Ireland: lessons from a country without guidelines. AB - BACKGROUND: Prostate specific antigen (PSA) testing is associated with increased prostate cancer (PCa) incidence. Ireland has no national guidelines on PCa screening and had the highest PCa incidence in Europe, 2006. AIMS: To investigate trends in PSA testing in Ireland. METHODS: Data on PSA tests, 1994-2005, was collated. RESULTS: Age-standardised rates of PSA testing increased 39 and 25% annually in men <50 and >or=50 years, respectively. Most tests were performed in men 50-69 years; 26 and 22% were performed in men <50 and >or=70 years, respectively. Baseline PSA tests peaked in 2004, at 16% of men. 83% of baseline tests were <4.0 ng/ml. Repeat testing increased with age and PSA level (P < 0.001); men <50 years and with levels <4.0 ng/ml had >1 tests in <21 months. PCa incidence increased 9% annually, 1994-2005. CONCLUSIONS: Uptake of PSA testing was rapid: increased use was simultaneous with increased PCa incidence. National guidelines are needed to manage this important public health issue. PMID- 19562408 TI - Establishing a temporal bone laboratory: considerations for ENT specialist training. AB - Cadaveric temporal bone dissection in a temporal bone laboratory is a vital component in training safe, competent otorhinolaryngologists. Recent controversies pertaining to organ retention have resulted in a more limited supply of temporal bones. Consequently, current trainees are dissecting far fewer bones than their consultants. We discuss the establishment of a temporal bone laboratory in the Department of Anatomy in the University College Cork, from the timely preparation and preservation of the tissue to its disposal. Comparisons are drawn between our experience and that of the United States training schemes. The temporal bone laboratory in Cork is the only one in existence in Ireland. The exposure and experience obtained by registrars rotating through Cork, has resulted in noticeable improvements in their operative abilities. The temporal bone laboratory remains a core component to training. It is hoped that this article may facilitate other units overcoming obstacles to establish a temporal bone laboratory. PMID- 19562409 TI - Psychiatric disorder treated in acute general hospitals: a comparison with psychiatric units and hospitals. AB - BACKGROUND: The extent to which psychiatric disorders are treated in acute general hospitals is unclear. AIMS: This study surveyed patients with psychiatric illness as their principal diagnosis treated in Irish Acute General Hospitals and compared their clinical characteristics with those in psychiatric settings. METHODS: The Hospital Inpatient Enquiry was used to determine the extent to which patients with psychiatric disorders were treated in the non-psychiatric wards of acute public general hospitals and the National Psychiatric Inpatient Reporting System employed to compare their diagnostic characteristics with those treated in psychiatric units and hospitals. CONCLUSIONS: There were substantial numbers of discharges for alcohol disorders from general hospitals wards and fewer, but not non-negligible, numbers for neurotic disorders, whereas schizophrenia and mania were almost exclusive to psychiatric units and hospitals. A significant number of eating disorders were treated in general hospitals with no increase over the period surveyed. PMID- 19562410 TI - Reversible acute cortical blindness associated with eclampsia in complete hydatidiform mole. AB - BACKGROUND: Eclampsia is rare in molar pregnancy. Sudden cortical blindness in eclampsia is an uncommon but very dramatic experience for the patient. Because of its rarity, blindness associated with eclampsia may pose a significant problem for the obstetrician. CASE REPORT: We describe cortical blindness and posterior reversible encephalopathy syndrome (PRES) complicating molar pregnancy-related eclampsia. The clinical presentation in our patient was consistent with PRES associated with eclampsia together with magnetic resonance imaging (MRI) findings. CONCLUSION: Preeclampsia and eclampsia are regarded as common causes of PRES, which is considered to be the result of vasogenic brain edema caused by a rapid raise in blood pressure. Clinical and imaging findings are usually reversible. Early diagnosis and elimination of possible causes are important in order to avoid permanent visual or brain injury. Imaging (especially MRI) should be carried out in eclamptic patients with visual disturbance in order to exclude other causes of blindness. PMID- 19562411 TI - An evaluation of the range and availability of intensive smoking cessation services in Ireland. AB - BACKGROUND: A review of smoking cessation (SC) services in Ireland is a necessary step in improving service planning and provision. AIMS: To assess the range and availability of intensive SC services in Ireland in 2006. METHODS: A survey of SC service providers in Ireland was conducted. Descriptive analysis and simple linear regression analysis was used. RESULTS: Response rate was 86.3% (63/73). All service providers surveyed are employing evidence-based interventions; the most common form of support is individual counselling with initial sessions averaging 40 min and weekly review sessions 20 min in duration. Reaching the recommended target of treating 5.0% of smokers does not seem feasible given the current distribution of resources and there appears to be regional differences in resource allocation. CONCLUSIONS: While intensive SC services are available in all four Health Service Executive Areas, it would appear that there is little uniformity or consistency countrywide in the scope and structure of these services. PMID- 19562412 TI - Detection of colo-rectal liver metastases: prospective comparison of contrast enhanced US, multidetector CT, PET/CT, and 1.5 Tesla MR with extracellular and reticulo-endothelial cell specific contrast agents. AB - BACKGROUND: To compare contrast-enhanced US (CE-US), multidetector-CT (MDCT), 1.5 Tesla MR with extra-cellular (Gd-enhanced) and intracellular (SPIO-enhanced) contrast agents and PET/CT, in the detection of hepatic metastases from colorectal cancer. MATERIALS AND METHODS: A total of 34 patients with colo-rectal adenocarcinoma underwent preoperatively CE-US, MDCT, Gd- and SPIO-enhanced MR imaging (MRI), and PET/CT. Each set of images was reviewed independently by two blinded observers. The ROC method was used to analyze the results, which were correlated with surgical findings, intraoperative US, histopathology, and MDCT follow-up. RESULTS: A total of 57 hepatic lesions were identified: 11 hemangiomas, 29 cysts, 1 focal fatty liver, 16 metastases (dimensional distribution: 5/16 < 5 mm; 3/16 between 5 mm and <10 mm; 8/16 >= 10 mm). Six of 34 patients were classified as positive for the presence of at least one metastasis. Considering all the metastases and those >= 10 mm, ROC areas showed no significant differences between Gd- and SPIO-enhanced MRI, which performed significantly better than the other modalities (P < 0.05). Considering the lesions <10 mm, ROC areas showed no significant differences between all modalities; however MRI presented a trend to perform better than the other techniques. Considering the patients, ROC areas showed no significant differences between all the modalities; however PET/CT seemed to perform better than the others. CONCLUSIONS: Gd- and SPIO-enhanced MRI seem to be the most accurate modality in the identification of liver metastases from colo-rectal carcinoma. PET/CT shows a trend to perform better than the other modalities in the identification of patients with liver metastases. PMID- 19562413 TI - Excretory urography: trends in clinical use and diagnostic yield. AB - OBJECTIVE: To assess for changes in clinical usage and diagnostic yield of excretory urography (EU) following the introduction of CT urography (CTU). MATERIALS AND METHODS: We retrospectively reviewed reports from 6313 EUs performed between July 1995 and February 2006. The specialty of the ordering physician and clinical indication for the study were recorded, as were any collecting system, ureter, or bladder abnormalities suspicious for urothelial malignancy identified on EU. The proportion of EUs ordered for each indication and the positivity rate for each finding were compared prior to and after 2000, when CTU was introduced. RESULTS: Demand for EU by all physicians has decreased threefold. Since 2000, there has been a decrease in the proportion of EUs performed for all indications: obstruction (P < 0.0001), stones (P < 0.0001), urothelial malignancy (P < 0.0001), pain (P < 0.0001), post-operative (P < 0.0001), trauma (P < 0.03), hematuria (P < 0.0001), and urinary tract infection (P < 0.0001). Proportional demand by urologists has not changed significantly (P = 0.105). For exams ordered by urologists, the positivity rate for intraluminal defects within the collecting system and bladder has decreased (P < 0.0001). For exams ordered by non-urologists, there has been no significant change in the positivity rate of urinary tract filling defects (P > 0.05). CONCLUSION: The number of excretory urograms has decreased dramatically, although the proportion of these studies ordered by urologists is unchanged. The positivity rate of EU findings suggesting urothelial malignancy in the collecting system and bladder has decreased, likely because, with the advent of CTU, urologists have changed their ordering patterns for some clinical indications. PMID- 19562414 TI - Evaluation of posttreatment response of hepatocellular carcinoma: comparison of ultrasonography with second-generation ultrasound contrast agent and multidetector CT. AB - We evaluated the ability of one-month follow-up contrast-enhanced ultrasound (CEUS) with second-generation contrast agent in monitoring radio frequency ablation (RFA) and transcatheter arterial chemoembolization (TACE) treatments of hepatocellular carcinoma (HCC). One-hundred forty-eight HCCs were studied using CEUS: 110 nodules were treated with RFA [41/110 RFA were performed using a pretreatment and an immediate postablation evaluation using CEUS (group 1); 69/110 using only US guidance (group 2)] and 38 nodules treated with TACE. For statistical analysis, McNemar test was used. Overall complete response was observed in 107/148 nodules (92/110 treated with RFA and 15/38 with TACE). A better rate of complete response was found in group 1 compared to group 2 (92.7% vs. 78.3%). In RFA treatment, CEUS showed a sensitivity of 83.3% and a specificity of 100% (diagnostic accuracy of 97%) using MDCT as reference standard with no statistical difference (p > 0.05). CEUS detected all cases of incomplete response in HCC treated with TACE using angiography as reference standard (diagnostic accuracy 100%). We recommend assessing residual intratumoral flow on CEUS during RFA procedure to determine the necessity of immediate additional treatment. In case of positive CEUS results, HCC treated with TACE should be considered still viable. PMID- 19562416 TI - Effects of predator and prey dispersal on success or failure of biological control. AB - Biological control, defined as the reduction of pest populations by natural enemies, is often a component of integrated pest management strategies. Augmentation of natural enemy numbers by planned releases is a common biological control method, the successes and failures of which have been extensively reviewed. The effectiveness of biological control is influenced by how populations of predators and prey (or hosts and parasitoids) disperse in patchy environments. Here, we address the question of whether such dispersal leads to beneficial or detrimental pest control outcomes by developing a simple predator prey model with constant releases of natural enemies in a two-patch environment. Theoretical and numerical results for all possible cases indicate that population dispersal has significant effects on the persistence of pests. For some ranges of dispersal rates or parameter space, dispersal is beneficial for pest control measures but this is not so for other ranges when it is detrimental. Therefore, knowledge of pest and natural enemy dispersal is crucial for understanding the effectiveness of biological control in a patchy environment. Finally, the model is generalised for multi-patch systems. PMID- 19562417 TI - Simplified negative pressure wound therapy in pediatric hand wounds. AB - Negative pressure wound therapy (NPWT) is commonly used as a bolster for skin grafts. The technique offers the benefit of negative pressure as well as reduced dressing changes. Skin grafting of the hand provides a unique challenge, and currently, the only commercially available NPWT hand dressings are adult-sized, precluding their use in small children. We present our custom NPWT "mitten" technique for use with skin grafts on the pediatric hand. PMID- 19562420 TI - Laparoscopic sleeve gastrectomy as revisional procedure for failed gastric banding and vertical banded gastroplasty. AB - BACKGROUND: The problem of revision of failed gastric banding (GB) and vertical banded gastroplasty (VBG) procedures has become a common situation in bariatric surgery. Laparoscopic sleeve gastrectomy (LSG) has been recently used to revise failed restrictive procedures. The objective of this study is to evaluate the results of LSG as revisional procedure for failed GB and VBG. METHODS: A prospective held database was questioned regarding patients' demographic, indication for revision, conversion to open surgery, morbidity, percentage of excess weight loss (%EWL), evolution of comorbidities, and need for a second procedure after LSG. RESULTS: Forty-one patients, 34 women and seven men with a mean age of 42 years (range 19 to 63 years) and a mean body mass index at 49.9 kg/m(2) (range 35.9-63 kg/m(2)), underwent laparoscopic conversion of GB (36 patients) and VBG (five patients) into LSG. Indication for revisional surgery was insufficient weight loss in all the cases. All procedures were completed laparoscopically. There was no mortality and five patients (12.2%) developed complications (high leak, one patient; intra-abdominal abscess, three patients; and complicated incisional hernia, one patient). At a mean follow-up of 13.4 months, %EWL is on average 42.7% (range 4-76.1%). Six patients had a second procedure (four had laparoscopic duodenal switch, one had laparoscopic Roux-en-Y gastric bypass, and one had laparoscopic biliopancreatic diversion). CONCLUSION: Conversion of GB and VBG into LSG is feasible and safe. LSG is effective in the short term with a mean %EWL of 42.7% at 13.4 months. Long-term results of LSG as revisional procedure are awaited to establish its efficacy in the long term. PMID- 19562421 TI - Adipose tissue of control and ex-obese patients exhibit differences in blood vessel content and resident mesenchymal stem cell population. AB - BACKGROUND: The normal function of white adipose tissue is disturbed in obesity. After weight loss that follows bariatric surgery, ex-obese patients undergo plastic surgery to remove residual tissues and it is not known whether their adipose tissue returns to its original state. The aim of this study was to compare the white adipose tissue composition of ex-obese with control patients with regard to blood vessels and resident mesenchymal stem cells (MSC). METHODS: Quantification of blood vessels was performed on histological sections of adipose tissue stained with hematoxylin and eosin and for von Willebrand antigen. MSC were induced to the adipogenic and osteogenic lineages by specific inductive culture media. Expression of PPARgamma2 was analyzed by reverse transcription polymerase chain reaction. RESULTS: Ex-obese adipose tissue showed a higher number (p = 0.0286) of small (107.3 +/- 22.0) and large (22.5 +/- 6.4) blood vessels, when compared to control patients (42.0 +/- 24.4 and 7.2 +/- 2.2, respectively) and they also occupied a larger area (control versus ex-obese, p = 0.0286). Adipose tissue MSC from both groups of patients expressed PPARgamma2 and were equally able to differentiate to the osteogenic lineage, but ex-obese MSC showed a higher adipogenic potential when induced in vitro (p < 0.05). CONCLUSIONS: The higher number of adipose tissue blood vessels in ex-obese patients explains the excessive bleeding observed during their plastic surgery. The presence of more committed cells to the adipogenic lineage may favor the easy weight regain that occurs in ex-obese patients. These results show that, after extensive weight loss, adipose tissue cell composition was not totally restored. PMID- 19562419 TI - Treatment of obesity in primary care practice in the United States: a systematic review. AB - OBJECTIVES: This review examines the results of randomized controlled trials in which behavioral weight loss interventions, used alone or with pharmacotherapy, were provided in primary care settings. DATA SOURCES: Literature search of MEDLINE, PubMed, Cochrane Systematic Reviews, CINAHL, and EMBASE (1950-present). Inclusion criteria for studies were: (1) randomized trial, (2) obesity intervention in US adults, and (3) conducted in primary care or explicitly intended to model a primary care setting. REVIEW METHODS: Both authors reviewed each study to extract treatment modality, provider, setting, weight change, and attrition. The CONSORT criteria were used to assess study quality. Due to the small number and heterogeneity of studies, results were summarized but not pooled quantitatively. RESULTS: Ten trials met the inclusion criteria. Studies were classified as: (1) PCP counseling alone, (2) PCP counseling + pharmacotherapy, and (3) "collaborative" obesity care (treatment delivered by a non-physician provider). Weight losses in the active treatment arms of these categories of studies ranged from 0.1 to 2.3 kg, 1.7 to 7.5 kg, and 0.4 to 7.7 kg, respectively. Most studies provided low- or moderate-intensity counseling, as defined by the US Preventive Services Task Force. CONCLUSIONS: Current evidence does not support the use of low- to moderate-intensity physician counseling for obesity, by itself, to achieve clinically meaningful weight loss. PCP counseling plus pharmacotherapy, or intensive counseling (from a dietitian or nurse) plus meal replacements may help patients achieve this goal. Further research is needed on different models of managing obesity in primary care practice. PMID- 19562423 TI - Implications of randomized studies of medical therapy vs revascularization for reducing rising costs of health care. PMID- 19562422 TI - Mortality and hospital stay after bariatric surgery in 2,167 patients: influence of the surgeon expertise. AB - BACKGROUND: Postoperative mortality is a rare event after bariatric surgery. The main goal of this study was to calculate the 30-day mortality rate postbariatric "open" surgery and the length of hospital stay of patients assisted by a health maintenance organization in Brazil. We also investigate their association with sex, age, BMI, preexisting comorbidities, and volume of procedures performed by surgeons. METHODS: A total of 2,167 patients who underwent RYGB between 01/2004 and 12/2007 were analyzed. The deaths and hospital stay were identified in the healthcare transactional database and the morbidity data in the preoperative medical audit records. Factors contributing to adverse outcomes were determined by multiple logistic regression analysis. RESULTS: The overall mortality rate was 0.64%, with a decreasing trend over the years. The median hospital stay was 3.1 days. In the multivariate analysis, both mortality and longer hospital stay were positively and significantly associated with age > 50 years, BMI > 50 kg/m(2), and surgeon volume of less than 20 bariatric surgeries/year. Presence of hypertension also increased the risk of longer hospital stay. Multivariate analysis showed that the 30-day mortality was six times higher in patients operated by professionals who performed less surgeries/year and longer hospital stay, four times more frequent. CONCLUSIONS: The 30-day mortality post-RYGB is similar to the rates found in developed countries and much lower than the rates found for patients assisted by the public health system in Brazil. In addition to age and clinical factors, the results suggest that mortality and longer hospital stay are strong and inversely related to surgeon's experience. PMID- 19562424 TI - Management of a patient with a large airway stent in situ. PMID- 19562425 TI - [Road traffic and the elderly]. PMID- 19562426 TI - [Alcohol, the aged and traffic]. AB - The roadworthiness of older people is affected by internal, neurological or orthopedic diseases and by the medication. These factors have an effect on alcohol tolerance. Therefore, alcoholization in elderly drivers needs specific attention. A retrospective evaluation of all alcoholized traffic participants > 60 years of age between 2003-2008 in Hamburg is described. The data analysis gives information about the alcohol level in various age groups, the resulting neurological/physiological deficits, chronic illnesses and the medications taken. The proportion of elderly people involved in a traffic delinquency has increased. However, the absolute number of old drunk drivers has decreased. Seniors are represented significantly less often concerning alcohol in traffic than other age groups. PMID- 19562427 TI - [Use of psychotropic substances by the elderly and driving accidents]. AB - In the context of a study for the Federal Highway Research Institute (Bundesanstalt fur Strassenwesen), a database containing almost 40,000 toxicological blood analysis results from drivers with various traffic offences (time frame 1998-2001) from throughout Germany was evaluated. The database contains the results of 25 laboratories in Germany performing toxicological blood analysis of driving offences. Of these laboratories, 23 gave information about their methodology for toxicological analysis by questionnaire. This database and the results from 10,000 toxicological blood analyses of traffic offences in Hamburg (time frame 2003-2008) were evaluated. The number of elderly driver cases, the spectrum of detectable substances, the type of offence and the medical findings were compared to the results of the total sample in the corresponding period. The number of traffic offences with elderly drivers has risen in recent years but to a much smaller degree than the total number of traffic offences. The relative frequency of detection decreased from 2% in 1998 to 1.1% in 2001 (Germany wide) and from 2.3% (2003) to 1.4% (2008) in Hamburg. In the group of elderly drivers, only 39% (43% in Hamburg) of the sample showed positive results for medical drugs or drugs of abuse (73.9% of all samples). The medical diagnosis on the symptomatic level of intoxication, which had been assigned by medical doctors after blood sampling, did not correlate with the actual presence of therapeutic drugs in the blood. This demonstrates the interactions of senso-motor and physical frailties with potential additive, substance-related effects when driving skills appear to be compromised in the elderly. Drugs of abuse were detected in 5.6% (10.6% in Hamburg) among the senior drivers. Benzodiazepines were detected in 24.5% (23% in Hamburg) of the cases with a high frequency of detection in traffic accidents compared to traffic offences without accident. The type of offence was given in 87 of benzodiazepine-positive cases (20 in Hamburg), 59 of them (11 in Hamburg) were traffic accidents. Keeping the increasing percentage of senior drivers in mind, the results show that most of the elderly drivers are very careful with medical therapy and driving. However traffic related safety advice including pharmaceutical side effects on driving ability given by general practitioners is important and necessary. PMID- 19562428 TI - [Driving ability and dementia]. AB - Physiological changes in cognitive competence in the elderly are clearly different to pathological changes such as dementia. Diagnosing dementia is not to be equated with absolute driving inability. During the course of dementia, the risk of an accident increases significantly. Detailed observation and regular checks serve to determine the appropriate time to hand in the driving licence. The basis of each decision should be a detailed history of the person and his or her immediate family. Attention should be paid to incorrect driving practise, unusual behaviour and uncertainty whilst driving as well as involvement in any accidents. An MMSE score below 24 or a CCT score up to 3 should result in an additional assessment being carried out, e.g. neuro-psychological tests, driving simulator or, if necessary on-road tests performed with specialists. Also helpful is the determination of functional status. Limitations in ADL or IADL function normally indicate an advanced stage of dementia. Due to the progression of the illness, the possibility of handing in the driving licence should be addressed as soon as possible. In this context, it is helpful to consider alternative forms of transportation. Individual wishes and needs must to be balanced against general road safety requirements. PMID- 19562429 TI - [Fitness Check for elder drivers]. AB - There is no compulsory and regular medical/psychological checkup for elderly drivers in Germany. In the near future, a growing danger to traffic safety is expected because of the increasing percentage of elderly people and increasing prevalence of geriatric diseases. To close this safety gap, TUV SUD Life Service GmbH has put into practice a voluntary consulting concept for elderly drivers (Fitness Check). The consultation is based on medical anamnesis and examination, psychological exploration of driving behaviour and achievement testing. If necessary, driving ability is tested during a car ride. Conditions, the course of and results of 51 checkups from 2006 to 2008 in Munich are described and conclusions discussed. PMID- 19562430 TI - [What roll does the sense of coherence in coping with Morbus Parkinson play?]. AB - PURPOSE OF THE STUDY: In this study the relevance of sense of coherence (SOC) for coping with an illness was examined in subjects with Parkinson's disease. According to Antonovsky's model (1997) the sense of coherence is an important resource when it comes to dealing with stressors. To take into consideration the integrated view of Parkinson patients, severity of the illness (UPDRS) was determined by the neurologist and tendency toward depression was recorded. METHOD: 51 patients with PD (mean age: 67.7; 43.1% female; 56.9% male) and 59 volunteers without any neurological illness (mean age: 65.7; 54.2% female; 45.8% male) took part in this study. The sample was recruited from the Neurological Department of the Medical University of Vienna. This quasi-experimental sample was assessed with standardized self-assessment questionnaires: FKV-LIS-SE, SOC Scale and GDS. Correlations, t-tests, U-tests, multivariate analyses of variance and linear regressions were used for calculation. RESULTS: Persons with PD were characterized by lower SOC (p<.01) and higher scores on depression (p<.01), compared to persons of the control group. Parkinson patients tend to use depressive and minimizing coping strategies (p<.01). In addition the study indicates an influence of SOC and tendency toward depression on coping (R(2)=0.43). Sense of coherence and coping strategies are independent of severity of illness, but there is a significant association between the duration of illness and active-problem focused coping. CONCLUSION: In general, sense of coherence correlates only with psychological variables, and not with physical variables. Results indicate the importance of SOC on effective coping. Therefore strengthening of SOC is important, especially in context with chronic neurological illness. Individual orientated analysis of resources should be implemented in every counselling interview, so that possibilities for activities of daily living and leisure can be developed. PMID- 19562431 TI - [Improvement of knowledge and attitudes towards depression and suicidality in geriatric caregivers: evaluation of an advanced training program]. AB - Depressive disorders rank among the most frequent mental disorders in the elderly. Especially in the elderly, depressive disorders are still underrecognized and not sufficiently treated. In view of the associated risks and complications, early and timely recognition of depressive disorders is essential to avoid exacerbation and chronic manifestation of the depressive symptoms and to antagonize the increased suicide risk as well as negative effects on concomitant somatic diseases. More than any other occupational group, geriatric caregivers have a key position in the contact with clients, relatives and treating primary care providers. Therefore, 3487 geriatric caregivers from 448 outpatient nursing services throughout Bavaria participated in 196 four-hour training courses about depressive disorders and suicidality in the elderly between March 2006 and August 2007. Aim of this activity was to reduce attitudes enclosed in a wall of prejudice and to teach specific knowledge about depression and suicidality in the elderly. This training was evaluated using specially tailored questionnaires which were completed immediately before and after the continuing education as well as three months later (at follow-up). The analysis included the individual changes of 473 participants in longitudinal sections. Pronounced knowledge deficits and reservations were found regarding pharmacotherapy with antidepressants and suicidality. In this context, significant changes in all categories were present. Especially pronounced effects were found for the attitudes towards depression and suicidality as well as treatability with antidepressants. These changes were stable, as indicated by the follow-up evaluation. PMID- 19562432 TI - [Body image and physical well-being in old age]. AB - The present paper examined the connection between a negative body image and body complaints in N = 788 persons above 60 years of age from the general population. From a psychosomatic view, it is to be expected that psychological adjustment problems to the aging body are accompanied by physical complaints. A cluster analysis resulted in a group of older people with a strongly negative body image and a high degree of body complaints. This group is mainly formed by women, persons in high age, elderly alone living and persons with little social support. A second cluster includes persons with a positive body image and few body complaints. These are mainly younger persons, men, persons with a high degree of social support and older people living in a partnership. A further cluster includes persons with a very negative body image, however, with few body complaints. These persons are younger and more often live in a partnership, compared to the elderly with negative body image and a high degree of body complaints. A fourth cluster represents persons with a low degree of a negative body image and few body complaints. Their vitality and physical well-being, however, are also low. The study confirms that body complaints are partly determined by the body image. Sex, life in partnership and social support are risk or protective factors, respectively. PMID- 19562433 TI - [Active and healthy living in old age--results from a representative survey of community-dwelling senior citizens in Hamburg]. AB - BACKGROUND: The majority of community-dwelling people 60 years and older are independent and live actively. However, there is little information about elderly persons' views on aging, health and health promotion. METHODS: Therefore, an anonymous, written questionnaire survey was performed in a representative sample of inhabitants from a section of the city of Hamburg, 60 years and older; 5 year intervals, 14 subsamples according to 7 age groups of females and males. RESULTS: Questionnaires from 950 participants (29% response) could be evaluated: mean age 71.5 years, 58% women, 34% living alone, 5% with professional healthcare needs as indicated by status according to German nursing care insurance. Senior citizens' positive attitudes towards aging and health were predominant: 69% of respondents felt young, 85% worried about loss of autonomy in old age. CONCLUSIONS: The results provide evidence indicating potential for improving health-promoting lifestyles in parts of the older population by evaluating and strengthening older persons' competencies and by considering their concerns seriously. These results provide valuable information for future plans in the public-health sector in the city of Hamburg where particular health-promoting actions for elderly persons will be considered. PMID- 19562436 TI - Pirouettes: the rotational play of wild chimpanzees. AB - A pirouette is a locomotor-rotational movement in which a young chimpanzee spins around in a (mostly) quadruped posture while advancing forward in a straight line. We addressed whether this behavior evolved as a practice of general athletic ability or of sexual selection. The former hypothesis would predict no sex differences in skill or the developmental process, while the latter would predict the opposite. Chimpanzees most likely master the pirouette around the time of weaning. We found no conspicuous sex differences in the developmental process or the number of rotations per bout of pirouettes, so the pirouette's main function may be to facilitate general athletic ability. Infants pirouetted regardless of the context of rest or travel, whereas juveniles and adolescents pirouetted primarily during travel. This is consistent with the survival strategy hypothesis, because juveniles and adolescents would be expected to display pirouettes to many watchers if this practice were sexually selected. However, the fact that males tend to pirouette faster than females and to pirouette even during adolescence suggests that sexual selection has some influence in shaping the evolution of the practice. Despite this, no conspicuous tendency was found for juveniles or adolescent chimpanzees to display pirouettes to opposite-sex individuals. More data on adolescent individuals are needed to definitively determine the role of sex differences in pirouetting behavior. PMID- 19562437 TI - Evaluation of dynamic row-action maximum likelihood algorithm reconstruction for quantitative 15O brain PET. AB - OBJECTIVE: A modified version of row-action maximum likelihood algorithm (RAMLA) using a 'subset-dependent' relaxation parameter for noise suppression, or dynamic RAMLA (DRAMA), has been proposed. The aim of this study was to assess the capability of DRAMA reconstruction for quantitative (15)O brain positron emission tomography (PET). METHODS: Seventeen healthy volunteers were studied using a 3D PET scanner. The PET study included 3 sequential PET scans for C(15)O, (15)O(2) and H (2) (15) O. First, the number of main iterations (N (it)) in DRAMA was optimized in relation to image convergence and statistical image noise. To estimate the statistical variance of reconstructed images on a pixel-by-pixel basis, a sinogram bootstrap method was applied using list-mode PET data. Once the optimal N (it) was determined, statistical image noise and quantitative parameters, i.e., cerebral blood flow (CBF), cerebral blood volume (CBV), cerebral metabolic rate of oxygen (CMRO(2)) and oxygen extraction fraction (OEF) were compared between DRAMA and conventional FBP. DRAMA images were post-filtered so that their spatial resolutions were matched with FBP images with a 6-mm FWHM Gaussian filter. RESULTS: Based on the count recovery data, N (it) = 3 was determined as an optimal parameter for (15)O PET data. The sinogram bootstrap analysis revealed that DRAMA reconstruction resulted in less statistical noise, especially in a low-activity region compared to FBP. Agreement of quantitative values between FBP and DRAMA was excellent. For DRAMA images, average gray matter values of CBF, CBV, CMRO(2) and OEF were 46.1 +/- 4.5 (mL/100 mL/min), 3.35 +/- 0.40 (mL/100 mL), 3.42 +/- 0.35 (mL/100 mL/min) and 42.1 +/- 3.8 (%), respectively. These values were comparable to corresponding values with FBP images: 46.6 +/- 4.6 (mL/100 mL/min), 3.34 +/- 0.39 (mL/100 mL), 3.48 +/- 0.34 (mL/100 mL/min) and 42.4 +/- 3.8 (%), respectively. CONCLUSION: DRAMA reconstruction is applicable to quantitative (15)O PET study and is superior to conventional FBP in terms of image quality. PMID- 19562438 TI - 11C-acetate for positron emission tomography imaging of clinical stage IA lung adenocarcinoma: comparison with 18F-fluorodeoxyglucose for imaging and evaluation of tumor aggressiveness. AB - BACKGROUND: To determine the usefulness of positron emission tomography (PET) with (11)C-acetate (AC) for imaging lung adenocarcinoma and evaluating its tumor aggressiveness, AC- and (18)F-fluorodeoxyglucose (FDG)-PET were compared. METHODS: One hundred and sixty-nine adenocarcinomas with clinical stage IA and 53 benign nodules were examined by both AC- and FDG-PET before surgery. The sensitivity and specificity for discriminating benign/adenocarcinoma were compared between AC- and FDG-PET. The AC and FDG uptakes were examined to determine the relationship with tumor aggressiveness, i.e., pathological tumor stage, lymphatic, vascular, or pleural involvement, and proliferative activity determined by Ki-67 staining score. RESULTS: While the sensitivity of AC-PET was significantly higher than FDG-PET for bronchioloalveolar carcinoma (BAC) and well differentiated (W/D) adenocarcinoma (p < 0.001 and 0.006, respectively), there was no significant difference for moderately or poorly differentiated adenocarcinoma. The specificity was not different between them. While FDG uptakes were significantly higher in tumors with pathological advanced stages or those with lymphatic, vascular and/or pleural involvements than in tumors with pathological stage IA or those without these tumor involvements (p = 0.04 to p < 0.001), AC uptake did not show significant differences between the respective sub groups except according to the tumor stage. While both AC and FDG uptakes showed a significant correlation with Ki-67 staining scores (p = 0.03 and p < 0.001, respectively), the correlation coefficient of former was lower than that of latter (p = 0.07). CONCLUSIONS: While AC-PET can image BAC and W/D adenocarcinoma with a higher sensitivity than FDG-PET, it cannot evaluate tumor aggressiveness of clinical stage IA lung adenocarcinoma as well as FDG-PET. PMID- 19562439 TI - Is thermal nociception only sensed by the capsaicin receptor, TRPV1? AB - Mammalian heat pain perception is well documented as a molecular event in the primary afferent neurons expressing TRPV1. Six types of thermo-TRPs were found, i.e., TRPV1-4, TRPM8 and TRPA1. The former TRPV1, 2 and TRPV3, 4 are sensitive to noxious heat and warmth, and the latter two are sensitive to cool or cold, respectively. We attempted to provide a hypothesis to explain the paradox in which TRPV1 knockout mice and capsaicin-pretreated mice with severe loss of small dorsal root ganglion (DRG) neurons behave normally to noxious heat. From the general view that TRPV1 is preferentially expressed in C-fibers responding to a moderate thermal threshold (>43 degrees C) and TRPV2 in Adelta-fibers to high threshold temperatures (>52 degrees C), the above phenomenon is perplexing. Woodbury et al. (J Neurosci 24:6410-6415, 2004) offered two pain transduction mechanisms, one being TRPV1/2-independent and the other TRPV1-dependent. The former detects noxious heat under normal conditions without the presence of TRPV1 or TRPV2, and the latter requires TRPV1 under pathophysiological conditions. Unidentified isolectin B4 (IB4)-positive but TRPV1-negative small neurons with a higher noxious heat threshold are feasible, because a spliced isoform of TRPV1 responsive to noxious heat (47 degrees C) but not responsive to either proton or capsaicin is present in human and rat sensory neurons. Thus, the IB4-positive but TRPV1-negative small sensory neurons must have a crucial role in the noxious heat response. PMID- 19562440 TI - Long-term effect of a thoracoscopic stapled bullectomy alone for preventing the recurrence of primary spontaneous pneumothorax. AB - PURPOSE: The purpose of this retrospective study was to determine the long-term efficacy of a stapled bullectomy without any symphysial procedures under videoassisted thoracoscopic surgery (VATS). METHODS: A total of 121 sides of 112 patients who underwent a stapled bullectomy alone for primary spontaneous pneumothorax were retrospectively reviewed. There were 48 sides of 45 patients who underwent VATS (VATS group) and 73 sides of 67 patients who underwent open surgery (thoracotomy group). RESULTS: There were 12 recurrences that occurred during the follow-up periods in the VATS group (24.5%), and 3 in the thoracotomy group (4.1%). The cumulative recurrence rates in the VATS group at 2 and 10 years after a bullectomy were 16.3% and 27.5%, whereas in the thoracotomy group the recurrence rates were 2.9% and 4.9%, respectively (P < 0.001). CONCLUSIONS: The long-term outcome of a VATS stapled bullectomy was unsatisfactory as a radical therapy for primary spontaneous pneumothorax. A symphysial procedure should therefore be added to VATS stapled bullectomy in order to prevent long-term postoperative recurrence. PMID- 19562441 TI - Crush syndrome due to drug-induced compartment syndrome: a rare condition not to be overlooked. AB - PURPOSE: The crush syndrome caused by drug-induced compartment syndrome (DCS) is a challenge for surgeons because it is regularly associated with potentially fatal complications. Drug-induced compartment syndrome can often be distinguished from other forms of compartment syndrome by the presence of severe rhabdomyolysis with kidney crush and severe postoperative complications such as local and generalized infection, persistent nerve damage, coagulopathy, and multiorgan failure. METHODS: In the past 15 years, eight prospectively documented, operatively managed, DCS with subsequent crush syndrome cases were recorded. RESULTS: All of the patients required renal replacement therapy. The creatine kinase (CK) values in the context of rhabdomyolysis reached an average of 86 (range 47-144) kU/l. The renal function recovered in all surviving patients. The analysis showed that the diagnosis of a DCS is usually made after an average of 13 h. It then took an average of an additional 7 h before a fasciotomy was performed. Six operational revisions were necessary. In three out of eight patients the extremities had to be amputated. CONCLUSIONS: In DCS the decision to open the compartment should be made immediately upon the clinical diagnosis. A protracted intensive phase is expected. The benefit to patients is closely associated with surgical wound debridement along with rigorous intensive therapy. PMID- 19562443 TI - A hidden danger of childhood trauma: bicycle handlebar injuries. AB - PURPOSE: To evaluate childhood bicycle handlebar injuries. METHODS: We evaluated retrospectively 14 patients who presented with bicycle handlebar injuries within a 3-year period. Bicycle injuries not caused by the handlebar were excluded. RESULTS: The mean age of the patients was 8.8 +/- 2 years (range, 5-12 years) and 79% were boys. The injuries comprised gastrointestinal perforation in 21%, traumatic abdominal hernia in 21%, and spleen laceration in 14%. The three children with intestinal perforation and the one with a penetrating abdominal injury underwent surgery, whereas the others were treated medically. An isolated traumatic abdominal hernia resolved spontaneously. There was no mortality. CONCLUSIONS: Although bicycle handlebar injuries occur at relatively low speeds, the transfer of energy from the end of the handlebar, with a small cross sectional area, to a small field leads to intra-abdominal injuries that are more severe than predicted. Thus, bicycle handlebar injuries should be considered as a serious intraabdominal injury until proven otherwise. PMID- 19562442 TI - Effects of hospital and surgeon case-volumes on postoperative complications and length of stay after esophagectomy in Japan. AB - PURPOSE: The exact relationship between larger caseload volume and lower morbidity following esophagectomy has not been established. This study investigates the effect of surgical volumes on reducing postoperative complications and length of stay after esophagectomy. METHODS: Patient and hospital data were collected electronically via a web-based questionnaire sent to surgeons in the Japan Surgical Society. Data were based on 642 patients treated with esophagectomy at 183 hospitals between November 1, 2006 and February 28, 2007. Multivariate analysis revealed that postoperative morbidity and length of stay regressed against hospital and surgeon volumes, patient characteristics, and details of the procedures. RESULTS: In a logistic regression model, esophagectomies by surgeons performing a high volume of operations (>100 cases; "high case-volume surgeons") were followed by a significantly lower rate of postoperative complications (odds ratio [OR], 0.49; 95% confidence interval (CI), 0.24-0.98, P = 0.04). In a proportional hazard model, high-volume surgeons reduced the length of stay significantly: the hazard ratio for medium casevolume surgeons (50-99 cases) was 1.53 [95% CI, 1.14-2.06, P = 0.00], whereas that for the highest case-volume surgeons was 1.34 [95% CI, 1.00-1.79, P = 0.05] vs the lowest case-volume surgeons. Neither postoperative complications nor length of stay were significantly associated with hospital volume. CONCLUSION: These findings indicate that morbidity after esophagectomy is more dependent on individual surgeon-specific skill than on hospital-based factors. PMID- 19562444 TI - Emergency colorectal resections in Asian octogenarians: factors impacting surgical outcome. AB - PURPOSE: The population in developing countries is aging and the number of octogenarians is expected to increase in the future at an alarmingly fast rate. This paper evaluates the surgical outcomes of emergency colorectal resections among Asian octogenarians. METHODS: We conducted a retrospective review of all octogenarians and older, who had an emergency colorectal resection performed between February 1996 to December 2001. RESULTS: Fifty-eight emergency colorectal resections were performed in patients with a median age of 83 years. The indications for surgery were colorectal cancer (74%) and complicated diverticular disease (12%). The most common presentation was an intestinal obstruction (72%). Fifty-one (88%) of the patients had comorbidities. Forty-five percent of patients had an American Society of Anesthesiologists (ASA) score of I and II, while 55% had a score of III and IV. Consultants performed 53% of the procedures. The mean surgical time was 156 min. Hartmann's procedure was the most common procedure performed (43%). There were 16 (28%) mortalities. The postoperative morbidity was 81%. The only factor impacting outcome was a high ASA score of III and IV. The median length of stay was 17.5 (range 3-108) days. CONCLUSIONS: The mortality and morbidity of emergency colorectal resections among Asian octogenarians are high and can be predicted by their ASA status. PMID- 19562445 TI - Comparison of Karydakis flap reconstruction versus primary midline closure in sacrococcygeal pilonidal disease: results of 200 military service members. AB - PURPOSE: There is a high incidence of sacrococcygeal pilonidal disease (SPD) among young males. Despite increasing data on the treatment of SPD, no standard treatment has yet been established. The goal of this study was to compare the Karydakis flap reconstruction with a primary midline closure (PMC) in the treatment of patients with SPD. METHODS: The study included 200 patients treated over a period of 30 months. After a power analysis, the study was run on a two directional basis (both retrospective reviews of patient charts and a telephone interview for prospective evaluation). The surgical findings, complications, recurrence rates, and degree of satisfaction based on a questionnaire were assessed. RESULTS: The duration of the surgery was longer in the Karydakis group. Complications were more frequently seen in the PMC group. The recurrence rate was 4.6% for the Karydakis group and 18.4% for the PMC group. Of patients who underwent the Karydakis operation, 70.8% were completely satisfied with the procedure, while only 32.6% of patients who underwent the PMC reported excellent satisfaction. A significantly higher number of patients in the Karydakis group recommended the same procedure to other patients with SPD. CONCLUSION: The Karydakis method may be a preferable option in the treatment of SPD, due to the low rate of recurrence and the promising satisfaction rates. PMID- 19562446 TI - Aloe versus silver sulfadiazine creams for second-degree burns: a randomized controlled study. AB - PURPOSE: Burn injury is associated with a high incidence of death and disability; yet its management remains problematic and costly. We conducted this clinical study to evaluate the efficacy of aloe vera cream for partial thickness burn wounds and compare its results with those of silver sulfadiazine (SSD). METHODS: Thirty patients with similar types of second-degree burns at two sites on different parts of the body were included in this study. Each patient had one burn treated with topical SSD and one treated with aloe cream, randomly. RESULTS: The rate of re-epithelialization and healing of the partial thickness burns was significantly faster in the site treated with aloe than in the site treated with SSD (15.9 +/- 2 vs 18.73 +/- 2.65 days, respectively; P < 0.0001). The sites treated with aloe were completely healed in less than 16 days vs 19 days for the sites treated with SSD. CONCLUSION: These results clearly demonstrated the greater efficacy of aloe cream over SSD cream for treating second-degree burns. PMID- 19562447 TI - Effects of N-acetylcysteine on regeneration following partial hepatectomy in rats with nonalcoholic fatty liver disease. AB - PURPOSE: To investigate the effect of N-acetylcysteine on regeneration following partial hepatectomy in rats with nonalcoholic fatty liver disease (NAFLD). METHODS: N-Acetylcysteine was given to seven rats with NAFLD (group 1); physiological saline was given to seven rats with NAFLD (group 2); and physiological saline was given to seven rats with a normal liver (group 3). We performed two-thirds hepatectomy in all rats and removed the remnant liver tissue 48 h later to measure the mitotic index (MI), proliferating cell nuclear antigen, glutathione (GSH), and malondialdehyde (MDA) levels. RESULTS: Mitotic index values were significantly higher in group 1 than in groups 2 and 3, and higher in group 3 than in group 2. Proliferating cell nuclear antigen values were significantly higher in group 1 than in group 2, but no significant difference was found in comparison with group 3. Glutathione values in group 1 were significantly higher than in group 2 and MDA values in group 1 were lower than in group 2. There was no significant difference between groups 1 and 3 in GSH and MDA values, in both the two-thirds hepatectomy and 48-h tissues. CONCLUSIONS: N Acetylcysteine enhanced regeneration after partial hepatectomy in rats with NAFLD. We believe that it exerted this effect through its influence on oxidative stress. PMID- 19562448 TI - Gefitinib and a ventriculo-peritoneal shunt to manage carcinomatous meningitis from non-small-cell lung cancer: report of two cases. AB - The prognosis of patients with carcinomatous meningitis from non-small-cell lung cancer (NSCLC) remains poor, and the available treatment options for the lung cancer do not relieve the severe symptoms of this sequela. We report the successful treatment of two cases of carcinomatous meningitis caused by NSCLC, using gefitinib and a ventriculo-peritoneal (V-P) shunt. The first patient was a 43-year-old woman with pT1N0M0 adenocarcinoma. Multiple brain and vertebral metastases were found 13 months after surgery. She had undergone gamma-knife radiosurgery for the brain metastases, radiotherapy for the vertebral metastases, and two regimens of systemic chemotherapy, before carcinomatous meningitis was diagnosed. She was given gefitinib, and then a V-P shunt was placed. She continued to take gefitinib and was free of subjected symptoms for 5 months until she died. The second patient was a 64-year-old woman with cT4N0M0 adenocarcinoma. After local chemotherapy using cisplatin and OK-432 for carcinomatosis pleuritis and two regimens of systemic chemotherapy, carcinomatous meningitis was detected. A V-P shunt was placed, and she was sequentially given gefitinib. At her 15-month follow-up, she was free of symptoms of carcinomatous meningitis. No adverse effects or shunt problems were detected in either patient. This therapeutic modality may liberate carcinomatous meningitis patients with severe symptoms from hospitalization and improve their quality of life. PMID- 19562449 TI - Paraplegia following the emergency surgical repair of a nonruptured symptomatic abdominal aortic aneurysm: report of a case. AB - This report presents an extremely rare case of paraplegia following emergency surgery for a nonruptured symptomatic abdominal aortic aneurysm. A 62-year-old man underwent an emergency surgical repair for a symptomatic nonruptured infrarenal abdominal aortic aneurysm. On postoperative day 2 paraplegia following spinal cord ischemia occurred at the T8 level. The site of the ischemia was situated too high for clamping to have caused this condition, unless the patient had a congenital anomaly in the blood supply to the spinal cord or it had been caused by the previously occluded great radicular artery, which was maintained by the collateral blood supply from the iliac circulation. PMID- 19562450 TI - Laparoscopic management of an obstructing granulocytic sarcoma of the jejunum causing intussusception in a nonleukemic patient: report of a case. AB - Granulocytic sarcoma is an extramedullary tumor of immature myeloid cells which is often a forerunner to the development of acute myelogenous leukemia. Granulocytic sarcoma of the gastrointestinal tract frequently involves the small intestine and often presents with abdominal pain and obstruction. Our patient presented with a proximal jejunal mass causing intussusception and obstruction. This type of manifestation has never before been reported. A laparoscopy-assisted resection of the affected portion of jejunum was performed for him. The initial pathological findings were high-grade non-Hodgkin's lymphoma; immunohistochemistry confirmed a diagnosis of granulocytic sarcoma. After a follow-up of 14 months, there was no evidence of leukemia. This condition is often mistaken for lymphoma and confirmation is necessary by immunohistochemistry. Chemotherapy is the treatment of choice and surgery is indicated only in the event of complications, such as bowel obstruction, bleeding, or perforation. The prognosis of granulocytic sarcoma is similar to that of myeloid leukemia. PMID- 19562451 TI - Idiopathic sclerosing encapsulating peritonitis--is a preoperative diagnosis possible? Report of three cases. AB - Sclerosing encapsulating peritonitis (SEP) is a rare cause of intestinal obstruction that is usually diagnosed during a laparotomy; however, a preoperative diagnosis is possible. Sclerosing encapsulating peritonitis can be classified as primary or idiopathic and secondary types and only about 70 cases of idiopathic type have been reported since it was first described. It is characterized by a total or partial encasement of the small bowel by a thick fibrotic membrane. This report presents a series of three cases in which a diagnosis of idiopathic SEP was made preoperatively based on the clinical features and radiological findings, which were confirmed by a laparotomy and histopathology. All of the cases were successfully managed by the excision of the membrane. This report demonstrates that based on the clinical features and radiological investigations, in the absence of other plausible etiologies for intestinal obstruction, it is possible to suspect a preoperative diagnosis of SEP, thereby preventing a "surprise" finding during a laparotomy and allowing for better management. PMID- 19562452 TI - Repair of a recurrent rectovaginal fistula using gluteal-fold flap: report of a case. AB - Various surgical procedures have been reported for the repair of a rectovaginal fistula, but a failure of the initial repair leads to difficulty in the second and later operations. This report presents the case of a 58-year-old woman with a recurrent rectovaginal fistula after a low anterior resection for rectal cancer who achieved a good outcome following a repair using a gluteal-fold flap. We therefore conclude that the transfer of a well-vascularized gluteal-fold flap may be a safe and reliable method to repair a rectovaginal fistula with a large diameter following repeated recurrence. PMID- 19562453 TI - Primary adenosquamous carcinoma of the colon: report of five cases. AB - Primary adenosquamous cell carcinomas (Ad-SCCs) of the colon and rectum are rare malignancies with a poor prognosis, in comparison to adenocarcinoma alone. Different roles of human papilloma virus (HPV) in its pathogenesis have been reported and the role of P16 in Ad-SCCs has not been explored. This report presents five cases of Ad-SCC of the colon to explore the clinicopathological features and the roles of P16, HPV 6/11, and 16/18. There was no confirmed evidence to support the relationship between the infection of HPV 6/11, 16/18, and pathogenesis of Ad-SCC of the colon. P16 overexpression was not related to HPV carcinogenesis and there might be another mechanism of P16 upregulation in Ad SCC of the colon. PMID- 19562454 TI - Intestinal malrotation associated with colon cancer in an adult: report of a case. AB - Intestinal malrotation is a congenital anomaly of intestinal rotation and fixation that generally becomes symptomatic during the neonatal period. However, it rarely occurs in adulthood, and synchronous colon cancer is even rarer. This report presents the case of a 45-year old man who suffered from intestinal malrotation associated with colon cancer. PMID- 19562455 TI - A left hepatectomy and caudate lobectomy combined resection of the ventral segment of the right anterior sector for hilar cholangiocarcinoma--the efficacy of PVE (portal vein embolization) in identifying the hepatic subsegment: report of a case. AB - This report presents a case of a left hepatectomy and a caudate lobectomy combined resection of the ventral segment of the right anterior sector for hilar cholangiocarcinoma using percutaneous transhepatic portal vein embolization (PVE). The patient was a 44-year-old man admitted to a local hospital with obstructive jaundice. He was diagnosed to have hilar cholangiocarcinoma and was referred to the hospital for further treatment. Cholangiography revealed stenosis of the left hepatic duct and the hilar bile ducts. The dorsal branch of the right anterior sector joined the right posterior branch and the tumor did not invade to the confluence of these branches. Arteriography and portography reconstructed by multidetector-raw computed tomography revealed the ventral branches of the right anterior sector, which separately diverged from the other right anterior branches. It was therefore necessary to perform a left hepatectomy and caudate lobectomy combined resection of the ventral segment of the right anterior sector to completely remove the tumor. Portal vein embolization was thus performed on the left portal vein and the ventral branches of the right anterior sector. Intraoperatively, when the hepatic artery was temporally clamped, the demarcation between the ventral segment and the dorsal segment of the right anterior sector could be clearly visualized. The planned surgery was performed safely. This case demonstrates that the utilization of PVE is useful for a difficult and intricate hepatectomy, which requires an accurate identification of a hepatic subsegment. PMID- 19562456 TI - Primary large gastrointestinal stromal tumor of the liver: report of a case. AB - A 30-year-old man was diagnosed to have a large intra-abdominal tumor in 1999. A histological examination of a resected liver specimen from an operation in 2002 revealed a gastrointestinal stromal tumor (GIST), diagnosed based on positive immunostaining for CD34 and c-kit. Two years after the operation, new lesions developed in the residual liver and the lesser curvature of the stomach. An immunohistological examination of both specimens showed the features of a GIST, thus matching those of the first histological examination of the liver GIST. While there were no mutations at exon 11 of c-kit in the liver GISTs resected in 2002 and 2004, the gastric lesion had a mutation at P577L (CCT to CTT) at exon 11. Therefore, the liver GIST and the gastric lesion were diagnosed to be independent. The patient already has survived for more than 9 years after receiving surgery three times and with administration of imatinib. PMID- 19562457 TI - How to handle a huge portosystemic shunt in adult living donor liver transplantation with a small-for-size graft: report of a case. AB - Little attention has been paid to a ligation of the spontaneous portosystemic shunt in adult living donor liver transplantation (LDLT). A 33-year-old Japanese man with cryptogenic liver cirrhosis accompanied by a huge splenorenal shunt underwent LDLT. Acute cellular rejection produced "to and fro" portal venous flow on postoperative day (POD) 10. Steroid bolus therapy reversed the rejection, but the recovery of the portal venous flow was incomplete and the recipient subsequently started to have episodes of encephalopathy. Angiography showed portal hypoperfusion and portal flow steal via a huge splenorenal shunt. The patient underwent a shunt occlusion on POD 58. Portography showed marked improvement of the portal hypoperfusion. The encephalopathy thereafter dramatically reversed and the patient was discharged with no complications related to shunt ligation on POD 110. This case suggested that a ligation of a huge portosystemic shunt should therefore be considered at the time of transplantation, even when a relatively small graft is implanted. PMID- 19562459 TI - What technique is suitable for laparoscopic suprapancreatic lymph node dissection? PMID- 19562458 TI - Experimental orthotopic lung transplantation model in rats with cold storage. AB - This report describes a new experimental procedure, a rat unilateral, orthotopic lung transplantation with cold storage, and evaluates its relevancy and reliability to study the early events during cold ischemia/reperfusion (I/R) injury. This model, using the cuff technique, does not require extensive training and is relatively easy to be established. The model can induce reproducible degrees of pulmonary graft injury including impaired gas exchange, proinflammatory cytokine upregulation, or inflammatory infiltrates, depending on the preservation time. The results are consistent with the previous clinical evidence, thus suggesting that this model is a valid and reliable animal model of cold I/R injury. PMID- 19562461 TI - Helicobacter pylori and gastric cancer. AB - Helicobacter pylori is now well known as an important pathogen related to the development of gastric cancer. However, some clinicians still doubt the causal association of H. pylori with the development of gastric cancer. To summarize the recent clinical data on the link between H. pylori and gastric cancer, we reviewed related articles published over the past 3 years, after the award of the Nobel Prize for Physiology or Medicine to Drs. J.R. Warren and B.J. Marshall for the first culture and isolation of H. pylori and the investigation of their relevance to peptic ulcer disease. This updated summary of the relationship between H. pylori and gastric cancer highlights the strong link between the organism and the development of gastric cancer, and suggests eradication of this bacterial infection as a possible prophylactic measure against the development of this lethal malignancy. At present, clinicians and researchers in the field emphasize the strong need for H. pylori eradication from the human stomach. PMID- 19562462 TI - A clinicopathological study of gastric stump carcinoma following proximal gastrectomy. AB - BACKGROUND: We aimed to clarify the frequency and clinicopathological characteristics of gastric stump carcinoma following proximal gastrectomy. METHODS: Three-hundred and sixteen patients who had undergone curative proximal gastrectomy over a 21-year period from January 1984 through December 2004 were reviewed. RESULTS: Gastric stump carcinoma was observed in 17 patients (5.4%). The time interval between the initial gastrectomy and the treatment of gastric stump cancer was within 5 years in 3 patients, within 5-10 years in 8, and after 10 years in 6. Treatment included endoscopic resection (n = 4), completion total gastrectomy of the remnant stomach (n = 11), pancreatoduodenectomy (n = 1), and nonsurgical resection (n = 1). Pathologically, 9 carcinomas were differentiated and 8 were undifferentiated. In a review of reconstruction methods associated with disease stage, stage I was found in 6 of the 7 patients with esophagogastrostomy or short-segment jejunal interposition. On the other hand, stage I was found in only 3, but stage II-IV was found in 7 of the 10 patients with reconstruction by double-tract or long-segment jejunal interposition; thus, the tumor was more likely to be detected at an advanced stage after long-segment interposition (P = 0.049). CONCLUSION: Gastric stump carcinoma following proximal gastrectomy occurred at a high frequency of 5.4% of initial resections. It is necessary to select a reconstruction method that facilitates postoperative endoscopic examination, as well as to follow up the patients after proximal gastrectomy in the long term for the early detection and early treatment of gastric stump carcinoma. PMID- 19562460 TI - Paclitaxel chemotherapy for the treatment of gastric cancer. AB - A comprehensive review of phase I and phase II clinical trials of paclitaxel and paclitaxel-containing chemotherapy regimens for advanced gastric cancer was performed. Response rates, median progression-free survivals, and median overall survivals were examined, together with the treatment regimens and the numbers of patients registered in each trial. Although paclitaxel monotherapy produced considerable improvement in tumor response and prognosis, combination doublet or triplet chemotherapy with fluoropyrimidines and/or platinum compounds showed better results than the paclitaxel monotherapy. With regard to the schedule of paclitaxel administration, weekly injection seemed to show less toxicity and better results than administration every 3 weeks. Adjuvant therapies, chemoradiation therapies, and paclitaxel treatment for gastric ascites were also investigated and are discussed. PMID- 19562463 TI - Serum interleukin-6 and -10 levels in patients with gastric cancer. AB - BACKGROUND: Interleukin-6 (IL-6) is known to be a multifunctional cytokine and IL 10 is an immunosuppressive factor. Both have been reported to be related to the disease prognosis in some human solid tumors. In the present study, we evaluated the clinical significance of preoperative serum IL-6 and IL-10 levels as new tumor markers in patients with gastric cancer (GC). METHODS: Preoperative serum samples from 90 patients with GC and 9 normal healthy volunteers were assayed. Levels of IL-6 and IL-10 were determined by enzyme-linked immunosorbent assay (ELISA). The clinical significance of serum IL-6 and IL-10 levels was evaluated and compared with serum carcinoembryonic antigen (CEA) levels and serum C reactive protein (CRP) levels in these patients. RESULTS: The serum level of IL-6 was significantly higher in the GC patients than in the healthy subjects. Serum IL-6 levels were strongly correlated with CRP levels, but did not correlate with CEA or carbohydrate antigen (CA) 19-9 levels. Serum IL-10 levels did not correlate with CEA, CA19-9, or CRP. Strong positive correlations between serum IL 6 levels and tumor size and tumor stage were observed. On the other hand, IL-10 did not correlate with such clinicopathological findings of tumors. However, high serum IL-10 levels were associated with a worse prognosis in the GC patients, independently of their tumor stage. CONCLUSION: These findings indicate that serum IL-6 may suggest gastric cancer progression. On the other hand, IL-10 may play an important role in host immunity and the prognosis of GC patients. PMID- 19562464 TI - New double-stapling technique for esophagojejunostomy and esophagogastrostomy in gastric cancer surgery, using a peroral intraluminal approach with a digital stapling system. AB - In the abdominal-transhiatal approach for resection of adenocarcinoma of the cardia or subcardia, and in laparoscopy-assisted total gastrectomy (LATG), the use of a circular stapling device has potential problems with the placement of the purse-string suture and insertion of the anvil of the instrument. We describe a new double-stapling technique for esophagojejunostomy and esophagogastrostomy, using a peroral intraluminal approach with a digital stapling system, a flexible shaft remote-control stapler - the Surg-ASSIST and Power Circular Stapler 21 mm (PCS). The overtube of the flexible shaft of the PCS is prepared with a nylon tie and secured to a nasogastric (NG) tube. The flexible shaft is manually advanced down the esophagus with guidance by pulling the NG tube from the abdominal cavity side. The trocar of the flexible shaft is removed from the stump of the abdominal esophagus and connected to the anvil and they are approximated; the stapler device is then fired to form a double-stapled esophagojejunostomy and esophagogastrostomy. Our peroral intraluminal approach does not require a suturing technique, and it can make anastomosis after resection for carcinoma of the esophagogastric junction and after LATG safe and simple. PMID- 19562465 TI - Left-sided approach for suprapancreatic lymph node dissection in laparoscopy assisted distal gastrectomy without duodenal transection. AB - Laparoscopy-assisted distal gastrectomy (LADG) with extended lymph node dissection has not yet been widely adopted for the treatment of gastric cancers because of the perceived complexity of the procedure. Suprapancreatic lymph node dissection is one of the most important and demanding procedures in this approach. The techniques of duodenal transection within the abdominal cavity or taping of the common hepatic or splenic artery had traditionally been adopted for suprapancreatic nodal dissection during open surgery. In 2005, we developed a new laparoscopic procedure to safely and simply perform suprapancreatic lymph node dissection in LADG. We introduced a left-sided approach for the dissection of lymph nodes in the left gastropancreatic fold, where the body of the stomach is turned over and lifted ventrally to expose the left gastropancreatic fold through the opened lesser sac, without duodenal transection, and the suprapancreatic lymph nodes are resected en bloc in reverse order, i.e., including the lymph nodes along the proximal splenic artery (station 11p), around the celiac artery (station 9), and along the common hepatic artery (station 8a). Between April 2005 and December 2007, a total of 391 patients with cT1,2 gastric cancer underwent this surgical approach. In all patients, surgery was completed safely with favorable outcomes; mean operating time was 239 min and mean blood loss was 63 ml. The complication rate was 4.6% (18/391); there were ten conversions (2.6%) and no mortality. The aim of the present study was to describe the surgical technique of our new approach for LADG with extended lymph node dissection and to evaluate the treatment outcomes achieved by this technique. PMID- 19562467 TI - Duodenal stump cancer after Billroth-II distal gastrectomy for gastric cancer. AB - Duodenal cancer is an uncommon neoplasm and it mostly arises from the periampullary area. However, metachronous or even recurrent cancer at the duodenal stump following Billroth II type distal gastrec tomy for gastric cancer is extremely rare and, to our knowledge, has not yet been reported. A 68-year-old man underwent Billroth II distal gastrectomy with D2 lymph node dissection for an advanced gastric cancer. At that time the tumor stage was T2bN3M0, with 44 of 78 retrieved lymph nodes showing metastasis. He was well without recurrence for 3 years; however, he visited our hospital because of the abrupt onset of dizziness and tarry stool. A polypoid tumor that bled easily when touched was found at the end of the afferent loop of the duodenal stump by gastrofiberscopic examination, and it was proven to be an adenocarcinoma by endoscopic biopsy. Fortunately, additional studies, including abdominal computed tomography and positron emission tomography-computed tomography, showed no other sites of recurrence. The patient underwent pancreaticoduodenectomy for local control of the recurrent tumor at the duodenal stump, and the pathologic findings, based on immunohistochemical staining, strongly suggested that the duodenal tumor was metachronous in nature rather than recurrent. PMID- 19562466 TI - Cronkhite-Canada syndrome complicated with huge intramucosal gastric cancer. AB - Cronkhite-Canada syndrome (CCS) is a rare nonhereditary disorder with gastrointestinal polyposis and associated ectodermal changes. This report documents a 59-year-old Japanese man with CCS who underwent a total gastrectomy for gastric tumors. The resected specimen showed a huge gastric adenocarcinoma with numerous polyps throughout the stomach. The cancer was pathologically limited to within the mucosa and its histological structure resembled that of hyperplasia in CCS polyps, which led us to suppose that the carcinoma had arisen from hyperplastic CCS polyps. These results urged us to study the phenotypic expression of mucins, which revealed MUC2(-) and MUC5AC(+) and supported the diagnosis of the tumor as a gastric-type well-differentiate adenocarcinoma. A literature search revealed that 32 gastric carcinomas which developed in patients with CCS were mostly limited to within the submucosa in spite of their huge sizes, and such cancer development in CCS polyposis is therefore not considered to be unusual. PMID- 19562468 TI - pH- and temperature-sensitive polymeric microspheres for drug delivery: the dissolution of copolymers modulates drug release. AB - Most pH-/temperature-responsive polymers for controlled release of drugs are used as cross-linked hydrogels. However, the solubility properties of the linear polymers below and above the lower critical solution temperature (LCST) are not exploited. Here, the preparation and characterization of poly (N isopropylacrylamide-co-methacrylic acid-co-methyl methacrylate) (poly (NIPAAm-co MA-co-MM)) and poly (N-isopropylacrylamide-co-acrylamide) (poly (NIPAAm-co-AAm)), known as "smart" polymers (SP), is reported. Both poly (NIPAAm-co-MA-co-MM) and poly (NIPAAm-co-AAm) display pH- and temperature-responsive properties. Poly (NIPAAm-co-MA-co-MM) was designed to be insoluble in the gastric fluid (pH = 1.2), but soluble in the intestinal fluid (pH = 6.8 and 7.4), at the body temperature (37 degrees C). Poly (NIPAAm-co-AAm) was designed to have a lower critical solution temperature (LCST) corresponding to 37 degrees C at pH = 7.4, therefore it is not soluble above the LCST. The solubility characteristics of these copolymers were exploited to modulate the rate of release of drugs by changing pH and/or temperature. These copolymers were solubilized with hydrophobic cellulose acetate butyrate (CAB) and vitamin B(12) (taken as a water soluble drug model system) in an acetone/methanol mixture and dispersed in mineral oil. By a progressive evaporation of the solvent, the liquid droplets were transformed into loaded CAB/SP microspheres. Differential scanning calorimetric studies and scanning electron microscopy analysis demonstrated that the polymeric components of the microspheres precipitated separately during solvent evaporation forming small microdomains. Moreover, vitamin B(12) was found to be molecularly dispersed in both microdomains with no specific affinity for any polymeric component of microspheres. The release of vitamin B(12) was investigated as a function of temperature, pH, and the CAB/SP ratio. PMID- 19562469 TI - Pharmacogenetics and population pharmacokinetics: impact of the design on three tests using the SAEM algorithm. AB - Pharmacogenetics is now widely investigated and health institutions acknowledge its place in clinical pharmacokinetics. Our objective is to assess through a simulation study, the impact of design on the statistical performances of three different tests used for analysis of pharmacogenetic information with nonlinear mixed effects models: (i) an ANOVA to test the relationship between the empirical Bayes estimates of the model parameter of interest and the genetic covariate, (ii) a global Wald test to assess whether estimates for the gene effect are significant, and (iii) a likelihood ratio test (LRT) between the model with and without the genetic covariate. We use the stochastic EM algorithm (SAEM) implemented in MONOLIX 2.1 software. The simulation setting is inspired from a real pharmacokinetic study. We investigate four designs with N the number of subjects and n the number of samples per subject: (i) N = 40/n = 4, similar to the original study, (ii) N = 80/n = 2 sorted in 4 groups, a design optimized using the PFIM software, (iii) a combined design, N = 20/n = 4 plus N = 80 with only a trough concentration and (iv) N = 200/n = 4, to approach asymptotic conditions. We find that the ANOVA has a correct type I error estimate regardless of design, however the sparser design was optimized. The type I error of the Wald test and LRT are moderatly inflated in the designs far from the asymptotic (<10%). For each design, the corrected power is analogous for the three tests. Among the three designs with a total of 160 observations, the design N = 80/n = 2 optimized with PFIM provides both the lowest standard error on the effect coefficients and the best power for the Wald test and the LRT while a high shrinkage decreases the power of the ANOVA. In conclusion, a correction method should be used for model-based tests in pharmacogenetic studies with reduced sample size and/or sparse sampling and, for the same amount of samples, some designs have better power than others. PMID- 19562470 TI - Comparison of spectroscopic and lasing properties of different types of sol-gel glass matrices containing Rh-6G. AB - Rhodamine-6G (Rh-6G) is embedded in sol-gel glass samples which have been prepared by three different methods namely: 1) using HCl as catalyst and glycerol as Drying Control Chemical Additive (DCCA), 2) using HCl as catalyst at 60 degrees C and drying at room temperature and 3) using HCl as catalyst at 60 degrees C and heated at 600 degrees C for 3 h. Comparative studies of spectroscopic and lasing properties of the three types of Rh-6G containing samples were carried out with the lapse of time upto 8 months. Photostability of Rh-6G containing sol-gel samples is measured in terms of half life under Nitrogen laser pumping as number of pulses of N2 laser necessary to reduce the dye laser intensity to 50% of the original value and value is 7500 pulses at 1.67 Hz rate. The best performance of Rh-6G, as far as its spectroscopic and lasing properties are concerned was found in third type of host matrices using HCl as catalyst at 60 degrees C and heated at 600 degrees C for 3 h. PMID- 19562471 TI - Tissue transglutaminase can be involved in airway inflammation of toluene diisocyanate-induced occupational asthma. AB - BACKGROUND: This study was conducted to evaluate whether tissue transglutaminase (tTG) may be involved in airway inflammation of toluene diisocyanate-induced occupational asthma (TDI-OA). METHODS: We enrolled 93 patients with TDI-OA, 177 asymptomatic exposed subjects, 43 patients with allergic asthma, and 70 unexposed normal controls. The prevalence of serum immunoglobulin G (IgG) to tTG in the TDI OA group (20.2%) was significantly higher than that in the three other groups (P < 0.001). RESULTS: TDI-OA patients with serum IgG to tTG had significantly lower methacholine PC(20) values (P < 0.02) and significantly higher prevalence of specific immunoglobulin E to vapor type TDI-human serum albumin conjugate (P < 0.01; r(2) = 0.411, P < 0.05). TDI exposure could increase tTG activity via reactive oxygen species (ROS) production, which was found to cross-link with cytokeratin 19 on immunoblot analysis. CONCLUSION: Therefore, TDI exposure may activate tTG via ROS-mediated mechanism in the airway epithelium leading to persistent airway inflammation in TDI-OA patients. PMID- 19562472 TI - siRNA knockdown of PD-L1 and PD-L2 in monocyte-derived dendritic cells only modestly improves proliferative responses to Gag by CD8(+) T cells from HIV-1 infected individuals. AB - INTRODUCTION: Due to their capacity to elicit and regulate immunity, dendritic cells (DCs) are important targets to improve vaccination. Knowing that programmed death-1 (PD-1) high virus-specific T cells become functionally exhausted during chronic exposure to human immunodeficiency virus-1 (HIV-1), the development of a therapeutic DC-based HIV-1 vaccine might include strategies that downregulate PD L1 and PD-L2 counter-receptors. METHODS: After showing that monocyte-derived DCs rapidly upregulated PD-L1 and PD-L2 expression upon maturation with a variety of stimuli, e.g., Toll-like receptor ligands and cytokines, we determined that PD-L1 and PD-L2 expression could be knocked down by electroporation of a single small interfering RNA (siRNA) sequence twice at the monocyte and immature stages of DC development. This knockdown approached completion and was specific and lasting for several days. RESULTS: We then added the PD-L1 and PD-L2 silenced monocyte derived DCs to peripheral blood mononuclear cells from HIV-1-infected individuals along with pools of 15-mer HIV-1 Gag p24 peptides. However, in cultures from six patients, there was only a modest enhancing effect of PD-L1 and PD-L2 silencing on CD8(+) T cell proliferative responses to the DCs. DISCUSSION: These findings suggest that, in monocyte-derived DCs, additional strategies than PD-L1 or PD-L2 blockade will be needed to improve the function of PD-1 high T cells. PMID- 19562473 TI - An integrated model of legal and moral reasoning and rule-violating behavior: the role of legal attitudes. AB - Legal socialization theory predicts that attitudes mediate the relation between legal reasoning and rule-violating behavior [Cohn, E. S., & White, S. O. (1990). Legal Socialization: A Study of Norms and Rules. New York: Springer-Verlag]. Moral development theory predicts that moral reasoning predicts rule-violating behavior directly as well as indirectly [Blasi, A. (1980). Bridging moral cognition and moral action: A critical review of the literature. Psychological Bulletin, 88, 1-45]. We present and test an integrated model of rule-violating behavior drawing on both theories in a longitudinal study of middle school and high school students. Students completed questionnaires three times during the course of 1 year at 6-month intervals. Legal and moral reasoning, legal attitudes, and rule-violating behavior were measured at times one, two, and three respectively. Structural equation models revealed that while moral and legal reasoning were directly and indirectly related to rule-violating behavior among high school students, legal reasoning bore no direct relation to rule-violating behavior among middle school students. The implications for an integrated model of reasoning and rule-violating behavior are discussed. PMID- 19562475 TI - Brief report: effects of cognitive behavioral therapy on parent-reported autism symptoms in school-age children with high-functioning autism. AB - This pilot study tested the effect of cognitive behavioral therapy (CBT) on parent-reported autism symptoms. Nineteen children with autism spectrum disorders and an anxiety disorder (7-11 years old) were randomly assigned to 16 sessions of CBT or a waitlist condition. The CBT program emphasized in vivo exposure supported by parent training and school consultation to promote social communication and emotion regulation skills. Parents completed a standardized autism symptom checklist at baseline and posttreatment/postwaitlist and 3-month follow-up assessments. CBT outperformed the waitlist condition at posttreatment/postwaitlist on total parent-reported autism symptoms (Cohen's d effect size = .77). Treatment gains were maintained at 3-month follow-up. Further investigation of this intervention modality with larger samples and broader outcome measures appears to be indicated. PMID- 19562474 TI - An ecological approach to understanding black-white disparities in perinatal mortality. AB - Despite appreciable improvement in the overall reduction of infant mortality in the United States, black infants are twice as likely to die within the first year of life as white infants, even after controlling for socioeconomic factors. There is consensus in the literature that a complex web of factors contributes to racial health disparities. This paper presents these factors utilizing the socioecological framework to underscore the importance of their interaction and its impact on birth outcomes of Black women. Based on a review of evidence-based research on Black-White disparities in infant mortality, we describe in this paper a missing potent ingredient in the application of the ecological model to understanding Black-White disparities in infant mortality: the historical context of the Black woman in the United States. The ecological model suggests that birth outcomes are impacted by maternal and family characteristics, which are in turn strongly influenced by the larger community and society. In addition to infant, maternal, family, community and societal characteristics, we present research linking racism to negative birth outcomes and describe how it permeates and is embedded in every aspect of the lives of African American women. Understanding the contribution of history to the various factors of life of Black women in the United States will aid in developing more effective policies and programs to reduce Black infant mortality. PMID- 19562478 TI - Examining temporal associations between perceived maternal psychological control and early adolescent internalizing problems. AB - The present study examined a) the associations between adolescent-reported maternal psychological control and self-reported internalizing problems one year later, while simultaneously examining the opposite direction of effects and b) the equivalence of these associations across gender. Participants were 479 10-to 14-year old adolescents (55% female) participating in two waves of a study. Results from multi-group, cross-lagged panel analyses indicated that although early adolescents' depressive symptoms and social anxiety predicted change in perceived maternal psychological control one year later, perceived maternal psychological control did not predict change in early adolescents' internalizing problems. Elevated levels of depressive symptoms were associated with increased levels of perceived maternal psychological control one year later, whereas elevated levels of social anxiety were associated with decreased levels. Aforementioned findings were consistent across boys' and girls' data. Study findings extend limited research on adolescent effects, demonstrating that early adolescents' internalizing problems play a role in perceived maternal psychological control. PMID- 19562477 TI - The unity and diversity of inattention and hyperactivity/impulsivity in ADHD: evidence for a general factor with separable dimensions. AB - To examine the unity and diversity of inattention and hyperactivity/impulsivity symptom domains of Attention-Deficit/Hyperactivity Disorder (ADHD) in a clinical sample of adolescents with ADHD. Parents and adolescents were administered a semi structured diagnostic interview, the Schedule for Affective Disorders and Schizophrenia for School-Age Children - Present and Lifetime Version (K-SADS-PL), to assess adolescent ADHD. Data from 201 parent interviews and 189 adolescent interviews were examined. Four potential factor structures for the 18 ADHD symptoms were tested using confirmatory factor analysis: two models with correlated factors and two bifactor models. A bifactor model with two specific factors best accounted for adolescent symptoms, according to both parent and adolescents' reports. Replication of these findings from behavioral rating scales completed for this sample by parents and teachers indicates that the findings are not method- or informant-specific. The results suggest that there is an important unitary component to ADHD symptoms and separable dimensional traits of Inattention and Hyperactivity/Impulsivity. PMID- 19562480 TI - Minimal axillary lymph node involvement in breast cancer has different prognostic implications according to the staging procedure. AB - It is still controversial whether the identification of micrometastases and isolated tumor cells in the axillary lymph nodes of patients with breast cancer has any prognostic value. We evaluated the prognostic role of isolated tumor cells and micrometastases in the axillary lymph nodes in 3,158 consecutive patients pT1-2 pN0-N1mi (with a single involved lymph node) and M0, referred to the Division of Medical Oncology after surgery performed at the European Institute of Oncology from April 1997 to December 2002. Median follow-up was 6.3 years (range 0.1-11 years). Sentinel lymph node biopsy (SLNB) and axillary lymph node dissection (ALND) were performed in 2,087 and 1,071 patients, respectively. A worse metastasis-free survival was observed for patients with micrometastatic disease compared to node-negative patients, if staged with ALND (log-rank P < .0001; HR: 3.17; 95% CI 1.72-5.83 at multivariate analysis), but not for patients who underwent SLNB (log-rank P = 0.36). The presence of a single micrometastatic lymph node is associated with a higher risk of distant recurrence as compared to node-negative disease only for patients undergoing ALND for staging purposes. Treatment recommendations for systemic therapy should not take into account the presence of a single micrometastatic lymph node identified during complete serial sectioning of sentinel node(s). PMID- 19562479 TI - Family and neighborhood fit or misfit and the adaptation of Mexican Americans. AB - In this study, a person-environment fit model was used to understand the independent and combined roles of family and neighborhood characteristics on the adjustment of adults and children in a sample of 750 Mexican American families. Latent class analysis was used to identify six qualitatively distinct family types and three quantitatively distinct neighborhood types using socioeconomic and cultural indicators at each level. The results showed that members of single parent Mexican American families may be particularly at-risk, members of the lowest-income immigrant families reported fewer adaptation problems if they lived in low-income neighborhoods dominated by immigrants, members of economically successful immigrant families may be more at-risk in integrated middle class neighborhoods than in low-income neighborhoods dominated by immigrants, and members of two-parent immigrant families appear to be rather resilient in most settings despite their low socioeconomic status. PMID- 19562482 TI - Light acclimation and HSO(3) (-) damage on photosynthetic apparatus of three subtropical forest species. AB - The effects of long-term (33 months) sun/shade acclimation and short-term (within 10 h) HSO(3) (-) treatment on leaf photosynthetic apparatus were investigated in three subtropical forest plants, Pinus massoniana, Schima superba, and Acmena acuminatissima. After 33 months' growth in two light environments (100 and 12% sunlight), rapid light curves (RLC), chlorophyll fluorescence imaging and chloroplast ultrastructures of three tested species were changed to different degrees. When leaf sections were immersed in 50 mM NaHSO(3) for 10 h, all the RLCs were lowered; chlorophyll fluorescence imaging was inclined to present warmer colors and imaging areas were decreased. However, changes in chloroplast ultrastructures differed from three species. Our results showed that the photosynthetic apparatus of a dominant species, A. acuminatissima, in the late succession stage of a subtropical forest in South China, was less sensitive to NaHSO(3) under both growing light intensities. Conversely, the chloroplasts of P. massoniana, the pioneer heliophyte species, were most susceptible to NaHSO(3). It is deduced that, SO(2) pollution may become as a factor to accelerate the succession of subtropical forest. PMID- 19562483 TI - Federal environmental legislation in the U.S. for protection of wildlife and regulation of environmental contaminants. AB - The U.S. has a long history of legislation to protect wildlife, beginning with the Lacey Act of 1900. There are now over 170 Federal laws that regulate environmental activities which may affect wildlife. Two important laws are the Pittman-Robertson Act enacted in 1937 that authorizes a tax for wildlife management and the Fish and Wildlife Coordination Act passed in 1958 whose primary purpose is conservation of fish and wildlife, both of which continue to provide significant funding for wildlife management. Modern environmental regulations began by passage of the National Environmental Policy Act in 1969, followed by the Clean Water Act, Superfund, and other laws to regulate pesticides and toxics and clean up contaminated sites. International conventions regulate sale, use and disposal of toxics and ocean dumping. These laws and conventions should protect wildlife from unintended consequences of global industrialization. PMID- 19562481 TI - Syntheses of mucin-type O-glycopeptides and oligosaccharides using transglycosylation and reverse-hydrolysis activities of Bifidobacterium endo alpha-N-acetylgalactosaminidase. AB - Endo-alpha-N-acetylgalactosaminidase catalyzes the release of Galbeta1-3GalNAc from the core 1-type O-glycan (Galbeta1-3GalNAcalpha1-Ser/Thr) of mucin glycoproteins and synthetic p-nitrophenyl (pNP) alpha-linked substrates. Here, we report the enzymatic syntheses of core 1 disaccharide-containing glycopeptides using the transglycosylation activity of endo-alpha-N-acetylgalactosaminidase (EngBF) from Bifidobacterium longum. The enzyme directly transferred Galbeta1 3GalNAc to serine or threonine residues of bioactive peptides such as PAMP-12, bradykinin, peptide-T and MUC1a when Galbeta1-3GalNAcalpha1-pNP was used as a donor substrate. The enzyme was also found to catalyze the reverse-hydrolysis reaction. EngBF synthesized the core 1 disaccharide-containing oligosaccharides when the enzyme was incubated with either glucose or lactose and Galbeta1-3GalNAc prepared from porcine gastric mucin using bifidobacterial cells expressing endo alpha-N-acetylgalactosaminidase. Synthesized oligosaccharides are promising prebiotics for bifidobacteria. PMID- 19562485 TI - Clozapine administration modifies neurotensin effect on synaptosomal membrane Na+, K+ -ATPase activity. AB - Na+, K+-ATPase is inhibited by neurotensin, an effect which involves the peptide high affinity receptor (NTS1). Neurotensin effect on cerebral cortex synaptosomal membrane Na+, K+-ATPase activity of rats injected i.p. with antipsychotic clozapine was studied. Whereas 3.5 x 10(-6) M neurotensin decreased 44% Na+, K+ ATPase activity in the controls, the peptide failed to modify enzyme activity 30 min after a single 3.0, 10.0 and 30.0 mg/kg clozapine dose. Neurotensin decreased Na+, K+-ATPase activity 40 or 20% 18 h after 3.0 or 5.6 mg/kg clozapine administration, respectively, and lacked inhibitory effect 18 h after 17.8 and 30.0 mg/kg clozapine doses. Results indicated that the clozapine treatment differentially modifies the further effect of neurotensin on synaptosomal membrane Na+, K+-ATPase activity according to time and dose conditions employed. Taken into account that clozapine blocks the dopaminergic D2 receptor, findings obtained favor the view of an interplay among neurotensinergic receptor, dopaminergic D2 receptor and Na+, K+-ATPase at synaptic membranes. PMID- 19562484 TI - Differential proteomics analysis of specific carbonylated proteins in the temporal cortex of aged rats: the deterioration of antioxidant system. AB - Oxidative stress plays a pivotal role in normal brain aging and various neurodegenerative diseases, including Alzheimer's disease (AD). Irreversible protein carbonylation, a widely used marker for oxidative stress, rises during aging. The temporal cortex is essential for learning and memory and particularly susceptible to oxidative stress during aging and in AD patients. In this study, we used 2-DE, MALDI-TOF/TOF MS, and Western blotting to analyze the differentially carbonylated proteins in the rat temporal cortex between 1-month old and 24-month-old. We showed that the carbonyl levels of ten protein spots corresponding to six gene products: SOD1, SOD2, peroxiredoxin 1, peptidylprolyl isomerase A, cofilin 1, and adenylate kinase 1, significantly increased in the temporal cortex of aged rats. These proteins are associated with antioxidant defense, the cytoskeleton, and energy metabolism. Several oxidized proteins identified in aged rat brain are known to be involved in neurodegenerative disorders as well. Our findings indicate that these carbonylated proteins may be implicated in the decline of normal brain aging process and provide insights into the mechanisms underlying age-associated dysfunction of temporal cortex. PMID- 19562486 TI - Factors associated with use of mental health services for depressed and/or suicidal youth aged 15-24. AB - Using data from a national survey, this study examined different factors and their association with mental health service use in youth aged 15-24 with depression and/or suicidality. Our sample consisted of 1,252 youth with depression and/or suicidality. In youth aged 15-18 years old, two factors influenced service use: (1) co-occurrence of depression and suicidality (OR 3.23, 95% CI 1.43, 7.32) and (2) chronic health condition (OR 2.44, 95% CI 1.22, 4.84). Several factors were associated with service use in youth aged 19-24 including gender (female) (OR 1.77, 95% CI 1.14, 2.76), and low income (OR 1.89, 95% CI 1.11, 3.23). The findings suggest differences in the factors associated with mental health service use between youth aged 15-18 and 19-24 with depression and/or suicidality. PMID- 19562488 TI - A microperfused incubator for tissue mimetic 3D cultures. AB - High density, three-dimensional (3D) cultures present physical similarities to in vivo tissue and are invaluable tools for pre-clinical therapeutic discoveries and development of tissue engineered constructs. Unfortunately, the use of dense cultures is hindered by intra-culture transport limits allowing just a few layer thick cultures for reproducible studies. In order to overcome diffusion limits in intra-culture nutrient and gas availability, a simple scalable microfluidic perfusion platform was developed and validated. A novel perfusion approach maintained laminar flow of nutrients through the culture to meet metabolic need, while removing depleted medium and catabolites. Velocity distributions and 3D flow patterns were measured using microscopic particle image velocimetry. The effectiveness of forced convection laminar perfusion was confirmed by culturing 700 microm thick neural-astrocytic (1:1) constructs at cell density approaching that of the brain (50,000 cells/mm(3)). At the optimized flow rate of the nutrient medium, the culture viability reached 90% through the full construct thickness at 2 days of perfusion while unperfused controls exhibited widespread cell death. The membrane aerated perfusion platform was integrated within a miniature, imaging accessible enclosure enabling temperature and gas control of the culture environment. Temperature measurements demonstrated fast feedback response to environmental changes resulting in the maintenance of the physiological temperature within 37 +/- 0.2 degrees C. Reproducible culturing of tissue equivalents within dynamically controlled environments will provide higher fidelity to in vivo function in an in vitro accessible format for cell-based assays and regenerative medicine. PMID- 19562487 TI - Non-invasive imaging of cardiac activation and recovery. AB - The sequences of activation and recovery of the heart have physiological and clinical relevance. We report on progress made over the last years in the method that images these timings based on an equivalent double layer on the myocardial surface serving as the equivalent source of cardiac activity, with local transmembrane potentials (TMP) acting as their strength. The TMP wave forms were described analytically by timing parameters, found by minimizing the difference between observed body surface potentials and those based on the source description. The parameter estimation procedure involved is non-linear, and consequently requires the specification of initial estimates of its solution. Those of the timing of depolarization were based on the fastest route algorithm, taking into account properties of anisotropic propagation inside the myocardium. Those of recovery were based on electrotonic effects. Body surface potentials and individual geometry were recorded on: a healthy subject, a WPW patient and a Brugada patient during an Ajmaline provocation test. In all three cases, the inversely estimated timing agreed entirely with available physiological knowledge. The improvements to the inverse procedure made are attributed to our use of initial estimates based on the general electrophysiology of propagation. The quality of the results and the required computation time permit the application of this inverse procedure in a clinical setting. PMID- 19562489 TI - Studies on the effect of water-soluble polymers on drug-cyclodextrin complex solubility. AB - The effect of complexation of irbesartan (IRB), a practically water-insoluble drug, with cyclodextrins in presence of different concentrations of water-soluble polymers (PEG 4000 and PVP K-90) on the dissolution rate of the drug has been investigated. Phase solubility studies were carried out to evaluate the solubilizing power of betaCD in association with water-soluble polymers towards IRB and to determine the apparent stability constant (K (S)) of the complexes. Improvement in K(S) value for ternary complexes (IRB-betaCD-polymers) clearly proved the benefit on the addition of water-soluble polymer to increase complexation efficiency. The dissolution rate of the drug from ternary systems containing PEG 4000 and PVP K-90 was higher as compared to the binary system. An optimum increase in the dissolution rate of the drug was observed at a polymer concentration of 5% w/w for PVP K-90 and 10% w/w for PEG 4000. DSC, FTIR, SEM, and XRD studies were carried out to characterize the complexes. PMID- 19562490 TI - Predictors of locating children participants in epidemiological studies 20 years after last contact: Internet resources and longitudinal research. AB - This study examines predictors of locating participants that were last contacted 20 years ago using public web-search directories, in order to facilitate longitudinal environmental health research. Participants (n = 3,202) resided in four distinct geographical neighborhoods in Hamilton, Ontario during childhood; they were between 15 and 17 years old when they were last contacted in 1986. Data used for tracing included available addresses, telephone numbers, given names, and parental names. Reverse and forward search strategies were used to retrieve updated contact details. 43% of the sample was traced using online directories. Following ethical approval, participants were contacted using traced data and 29% of the original cohort was located. Predictors of locating participants were: availability of paternal names, being traced to original addresses or telephone numbers, gender (male), relatively higher socioeconomic status in childhood, and not being exposed to smoking in childhood. Where participants resided in childhood was not a significant predictor of locating participants. Although 13% of the sample was traced using forward search by name, only 4% were located. For participants traced to available addresses or telephone numbers, the difference between the proportions of traced and located participants was <3%. Prospective studies on children may benefit from including the listed names that pertain to each child's telephone number and full parental names at recruitment, thereby increasing the likelihood of locating participants using Internet resources. Integrating the use of Internet-based public directories for cohort reconstruction can reduce financial costs related to follow-up for longitudinal research. PMID- 19562491 TI - Characterization of ochratoxin A-induced apoptosis in primary rat hepatocytes. AB - The main target organ of the mycotoxin ochratoxin A (OTA) in mammals is the kidney but OTA has also been shown to be hepatotoxic in rats and to induce tumors in mouse liver. Even at very low concentrations, OTA causes perturbations of cellular signaling pathways as well as enhanced apoptosis. OTA has been extensively studied in kidney cell systems. Since this substance also affects liver health, we focused our work on apoptosis-related events induced by OTA in primary rat hepatocytes. We performed pathway-specific polymerase chain reaction arrays to assess the expression of genes involved in apoptosis. Treatment with 1 microM OTA for 24 h caused marked changes in apoptosis-related gene expression. Genes as apaf1, bad, caspase 7, polb (DNA polymerase beta, performs base excision repair), and p53, which are marker genes for DNA damage, were upregulated. FAS and faslg were also markedly induced by treatment with OTA. Treatment of hepatocytes with OTA led to a concentration-dependent inhibition of protein biosynthesis. Apoptosis-inducing factor was released from mitochondria following OTA treatment; the mycotoxin induced the activity of caspases 8, 9, and 3/7 and caused chromatin condensation and fragmentation. Caspase inhibition led to a significant but not complete reduction of OTA-induced apoptosis. Our data suggest that not only OTA leads to p53-dependent apoptosis in rat hepatocytes but it also hints to other mechanisms, independent of caspase activation or protein biosynthesis, being involved. PMID- 19562492 TI - Smoking and aggressive prostate cancer: a review of the epidemiologic evidence. AB - Although tobacco use has been recognized as one of the leading causes of cancer morbidity and mortality, a role of smoking in the occurrence of prostate cancer has not been established. However, evidence indicates that factors that influence the incidence of prostate cancer may differ from those that influence progression and fatality from the disease. Thus, we reviewed and summarized results from prospective cohort studies that assessed the relation between smoking and fatal prostate cancer risk, as well as epidemiological and clinical studies that focused on aggressive behavior in prostate cancer, such as poorer survival, advanced stage, or poorer differentiation at diagnosis. The majority of the prospective cohort studies showed that current smoking is associated with a moderate increase of ~30% in fatal prostate cancer risk compared to never/non smokers. This association is likely to be an underestimate of the effect of smoking because most studies had a single assessment of smoking at baseline and long follow-up times, and the association was considerably stronger in some sub groups of heaviest smokers, or when smoking was assessed in a relatively short period (within 10 years) prior to cancer mortality. Using aggressive behavior of prostate cancer as outcome, current smoking was associated with significantly elevated risk, ranging from around twofold to threefold or higher. Although alternative explanations, such as publication bias, residual confounding, screening bias, and the influence of smoking-related comorbidities cannot be ruled out entirely, these findings suggest that smoking is associated with aggressive behavior of prostate cancers or with a sub-group of rapidly progressing prostate cancer. Based on evidence presented in this review, cigarette smoking is likely to be a risk factor for prostate cancer progression and should be considered as a relevant exposure in prostate cancer research and prevention of mortality from this cancer. PMID- 19562493 TI - Influence of tumor stage, symptoms, and time of blood draw on serum concentrations of organochlorine compounds in exocrine pancreatic cancer. AB - BACKGROUND: Knowledge is scant on the relationships between pathophysiologic processes common during cancer progression and changes in blood concentrations of organochlorine compounds (OCs). OBJECTIVE: To analyze the influence of tumor stage, cancer symptoms, and time of blood extraction on serum concentrations of OCs in exocrine pancreatic cancer (EPC). METHODS: Subjects were 144 incident cases of EPC prospectively recruited in eastern Spain. Blood was drawn and face to-face interviews with patients were conducted during hospital admission. Information on signs and symptoms was obtained from medical records and patient interviews. OCs were analyzed by high-resolution gas chromatography with electron capture detection. General linear models were applied to analyze log-transformed OCs corrected for total lipids. RESULTS: Lower concentrations of six of the seven OCs analyzed (p,p'-DDE, three polychlorinated biphenyls, hexachlorobenzene, and beta-hexachlorocyclohexane) were observed in patients with cholestatic syndrome (jaundice, hypocholia, and choluria). The constitutional syndrome increased only p,p'-DDT. The lowering effect of the cholestatic syndrome was stronger than the increasing effect of the constitutional syndrome (fatigue, anorexia, and weight loss), except for p,p'-DDT. When symptoms were considered, stage had only weakly inverse relationships with OC levels. The effects of symptoms on p,p'-DDE, p,p' DDT, and the three PCBs remained significant after adjusting by the interval from blood extraction to first symptom of EPC, and even when further adjusting by stage. CONCLUSIONS: Restriction or adjustment by stage and timing of blood draw may be insufficient to prevent biases associated with cancer progression. Symptoms may enable investigators to assess disease-induced changes in lipophilic exposure biomarkers. PMID- 19562494 TI - Smoking attenuates the negative association between carotenoids consumption and colorectal cancer risk. AB - OBJECTIVES: Consumption of vegetables and fruits, physical activity, obesity and caloric intake are all strongly related to the risk of colorectal cancer (CRC). The association between dietary intake of carotenoids from vegetables/fruits and risk of CRC in the context of cigarette smoking was studied in a nutritionally diverse population. METHODS: The study included 1,817 age sex residence-matched case-control pairs from a population-based study in Northern Israel. Data were acquired by food-frequency questionnaire. Individual intake of carotenoid isomers was calculated using an Israeli food content database. Odds ratios (ORs) were calculated using conditional logistic regression models adjusted for known risk factors. RESULTS: Strong inverse associations were found with consumption of 9 cis-beta-carotene (OR = 0.35, 0.26-0.47), all-trans-beta-carotene (OR = 0.58, 0.44-0.76), cis-beta-cryptoxanthin (OR = 0.67, 0.50-0.90), all-trans-zeaxanthin (OR = 0.64, 0.48-0.86), and lutein (OR = 0.74, 0.57-0.96). Lycopene (OR = 2.22, 1.71-2.89) and all-trans-beta-cryptoxanthin (OR = 2.01, 1.48-2.73) were associated with increased risk of CRC. Inverse associations of most carotenoids with CRC, demonstrated in non-smokers, were much attenuated or reversed in past or current smokers with a highly significant interaction term. CONCLUSIONS: Consumption of most dietary carotenoids was found to be strongly associated with reduced risk of CRC. However, smoking significantly attenuated or reversed this observed protective effect on CRC occurrence. Smokers should be advised that smoking also hampers the potential health promoting effects of high fruit and vegetable consumption. PMID- 19562495 TI - Land use patterns and urbanization in the holy city of Varanasi, India: a scenario. AB - Rapid urbanization and increasing land use changes due to population and economic growth in selected landscapes is being witnessed of late in India and other developing countries. The cities are expanding in all directions resulting in large-scale urban sprawl and changes in urban land use. The spatial pattern of such changes is clearly noticed on the urban fringes or city peripheral rural areas than in the city center. In fact, this is reflected in changing urban land use patterns. There is an urgent need to accurately describe land use changes for planning and sustainable management. In the recent times, remote sensing is gaining importance as vital tool in the analysis and integration of spatial data. This study intends to estimate land use pattern in a planned and unplanned urban setup and also to analyze the impact of change in land use pattern in the Varanasi urban environment. The results indicate that the planned urban setup had a higher tree cover to that of unplanned area in the Varanasi City, although a considerable disparity existed within the planned urban setups. The results emphasize the need to critically review concepts of urban planning and give more consideration to the preservation and management of urban tree cover/greenspace. PMID- 19562496 TI - Temporal evolution of the main processes that control indoor pollution in an office microenvironment: a case study. AB - The aim of this study is to examine the relative contribution of the outdoor concentration, the ventilation rate, the geometric characteristics of the indoor environment (i.e., extent of indoor surfaces and indoor volume), the deposition, and chemical reactions to the indoor air quality of the office microenvironment. For this case study, the NO, NO2, and O3 concentrations indoors and outdoors and TVOCs and CO2 concentrations indoors were measured in an office microenvironment in Athens, Greece, that was ventilated both naturally and mechanically. The calculated ventilation and loss rates and the measured outdoor concentrations of NO, NO2, and O3 were set as input to Multi-chamber Indoor Air Quality Model in order to study the temporal variation of the indoor NO, NO2, and O3 concentrations. Results showed that when the ventilation rate and outdoor concentration are high, the relative contribution of the transport process contributes significantly, while the chemical process depends on the contemporary interplay between the indoor O3, NO, and NO2 concentrations and lighting levels. The significance of each process was further examined by performing sensitivity tests, and it was found that the most important parameters were the deposition velocities, the UV infiltration rates (which determines the indoor chemical reaction rates), the ventilation rates, and the filtration (when a mechanical ventilation system is used). The effect of the hydrocarbon chemistry was not significant. PMID- 19562497 TI - Hematology of great sturgeon (Huso huso Linnaeus, 1758) juvenile exposed to brackish water environment. AB - The effect of environmental salinity on hematological parameters of great sturgeon Huso huso juveniles was studied. Five-month-old juveniles (mean body weight 28.3 +/- 2.1 g) were subjected to 0, 3, 6, 9, and 12 ppt salinities. The hematological parameters were assessed after a period of 20 days rearing at these salinities. After transfer from fresh water to brackish water, red blood cells, hematocrit, haemoglobin and mean corpuscular haemoglobin decreased, but mean corpuscular volume increased. Mean corpuscular haemoglobin concentration, white blood cells, monocyte counts, and eosinophil counts showed no significant variations with increase in environmental salinity. An increase was found in lymphocyte counts according to the increase of salinity from 0 to 12 ppt, while the fresh water control group maintained basal levels. Decrease in neutrophil counts was observed in great sturgeon with increase in environmental salinity. These data show significant effect of salinity on the blood parameters of great sturgeon. PMID- 19562498 TI - Plastid transformation in eggplant (Solanum melongena L.). AB - We have developed a method for plastid transformation in eggplant (Solanum melongena L.), a solanaceous plant species. Plastid transformation in eggplant was achieved by bombardment of green stem segments with pPRV111A plastid expression vector carrying the aadA gene encoding aminoglycoside 3'' adenylyltransferase. Biolistic delivery of the pPRV111A plasmid yielded transplastomic plants at a frequency of two per 21 bombarded plates containing 25 stem explants each. Integration of the aadA gene in the plastome was verified by PCR analysis and also by Southern blotting using 16S rDNA (targeting sequence) and the aadA gene as a probe. Transplastomic expression of the aadA gene was verified by RT-PCR. The development of transplastomic technology in eggplant may open up exciting possibilities for novel gene introduction and expression in the engineered plastome for agronomic or pharmaceutical traits. PMID- 19562499 TI - Prevalence and antibiotic susceptibility of thermophilic Campylobacter isolates from free range domestic duck (Cairina moschata) in Morogoro municipality, Tanzania. AB - Prevalence and antibiotic susceptibility of thermophilic Campylobacter isolated from free-ranging ducks was determined in Morogoro Municipality, Tanzania. Ninety intestinal contents from ducks were screened for thermophilic Campylobacter using Skirrow's protocol. Of the Campylobacter jejuni isolates, 50 were tested for sensitivity to 12 antibiotics. Overall prevalence of thermophilic Campylobacter was 80%. The prevalence of Campylobacter in adult ducks (91.3%) was significantly (p < 0.05) higher than ducklings (68.2%). The isolation rate of C. jejuni (81.9%) was significantly (P < 0.001) higher than C. coli (18.1%). All C. jejuni isolates were susceptible to streptomycin, nitrofurantoin and amikacin. Forty eight percent, 74% and 82% of isolates were resistant to cefuroxime sodium, tetracycline and ampicillin respectively. Between 20-50% of isolates were resistant to erythromycin, gentamicin, cloxacillin and amoxicillin. Norfloxacin and ciprofloxacin had lower C. jejuni resistance of 10% and 16% respectively. C. jejuni isolates from adult ducks showed significantly higher rates of resistance (p < 0.05) to most antibiotics than did duckling isolates. High prevalence of thermophilic Campylobacter in ducks could be of public health significance in Morogoro municipality. The observed multidrug resistance in this study poses a threat of transfer of antibiotic resistance to human pathogens because of the close contact between ducks and human. PMID- 19562501 TI - IgM monoclonal component associated with type I Gaucher disease resolved after enzyme replacement therapy: a case report. AB - The frequency of monoclonal gammopathy of undetermined significance (MGUS) is higher in patients with type I Gaucher disease (GD I) than in the general population. Although enzyme replacement therapy is effective in the control of the disease, its effect on MGUS is still controversial. We present the case of a 65-year-old woman with extensive GD I associated with IgM MGUS, in whom enzyme replacement therapy succeeded in eradicating the monoclonal component. This observation further supports the idea that enzyme replacement therapy decreases the chronic antigenic stimulation responsible for gammopathies in Gaucher disease. PMID- 19562500 TI - Current issues around the pharmacotherapy of ADHD in children and adults. AB - BACKGROUND: New drugs and new formulations enter the growing market for ADHD medication. The growing awareness of possible persistence of ADHD impairment beyond childhood and adolescence resulting in increased pharmacotherapy of ADHD in adults, is also a good reason for making an inventory of the what is generally known about pharmacotherapy in ADHD. AIM: To discuss current issues in the possible pharmacotherapy treatment of ADHD in children, adolescents and adults with respect to the position of pharmacotherapy in ADHD treatment guidelines, the pharmacoepidemiological trends, and current concerns about the drugs used. METHODS: A search of the literature with an emphasis on the position of pharmacotherapy in ADHD treatment guidelines, the pharmacoepidemiological trends, and current concerns about the drugs used in pharmacotherapy. RESULTS: According to the guidelines, the treatment of ADHD in children consists of psychosocial interventions in combination with pharmacotherapy when needed. Stimulants are the first-choice drugs in the pharmacological treatment of ADHD in children despite a number of well known and frequently reported side effects like sleep disorders and loss of appetite. With regard to the treatment of adults, stimulant treatment was recommended as the first-choice pharmacotherapy in the single guideline available. Both in children and adults, there appears to be an additional though limited role for the nonadrenergic drug atomoxetine. The increase of ADHD medication use, in children, adolescents and in adults, can not only be interpreted as a sign of overdiagnosis of ADHD. Despite the frequent use of stimulants, there is still a lack of clarity on the effects of long-term use on growth and nutritional status of children. Cardiovascular effects of both stimulants and atomoxetine are rare but can be severe. The literature suggests that atomoxetine may be associated with suicidal ideation in children. CONCLUSION: Although pharmacotherapy is increasing common in the treatment of ADHD in both children and adults, there are still a lot of questions about side effects and how best to counter them. This suggests an important role for close monitoring of children and adults treated with stimulants or atomoxetine. PMID- 19562502 TI - Continuous infusion of enzyme replacement therapy is inferior to weekly infusions in MPS I dogs. AB - Intravenous enzyme replacement therapy with recombinant human alpha-L-iduronidase (rhIDU) is used weekly to treat mucopolysaccharidosis (MPS) I. We tested continuous administration of rhIDU at two dosing levels (0.58 mg/kg per week and 2 mg/kg per week) in MPS I dogs, and compared the efficacy of continuous infusion with the clinically used 0.58 mg/kg weekly three-hour infusion. Peak plasma concentrations of rhIDU were much higher in weekly-treated dogs (mean 256 units/ml) than steady-state concentrations in dogs treated with continuous infusion (mean 1.97 units/ml at 0.58 mg/kg per week; 8.44 units/ml at 2 mg/kg per week). Dogs receiving continuous IV rhIDU, even at a higher (2 mg/kg per week) dose, had consistently lower iduronidase levels in tissues than dogs receiving a weekly (0.58 mg/kg per week) dose. GAG storage was also less improved by continuous intravenous infusion. Adverse events were similar in all dosing groups. We found that continuous administration of 2 mg/kg per week rhIDU to MPS I dogs was insufficient to achieve GAG storage reduction comparable to 0.58 mg/kg weekly dosing. PMID- 19562503 TI - Orotic aciduria and uridine monophosphate synthase: a reappraisal. AB - Three subtypes of hereditary orotic aciduria are described in the literature, all related to deficiencies in uridine monophosphate synthase, the multifunctional enzyme that contains both orotate: pyrophosphoryl transferase and orotidine monophosphate decarboxylase activities. The type of enzyme defect present in the subtypes has been re-examined by steady-state modelling of the relative outputs of the three enzymic products, uridine monophosphate, urinary orotic acid and urinary orotidine. It is shown that the ratio of urinary outputs of orotidine to orotate provides a means of testing for particular forms of enzyme defect. It is confirmed that the type I defect is caused by loss of uridine monophosphate synthase activity. Cells and tissue of type I cases have a residual amount of activity that is qualitatively unchanged: the relative rates of the transferase and decarboxylase do not differ from those of wild-type enzyme. The single claimed case of type II, thought to be due to specific inactivation of orotidine monophosphate decarboxylase, is shown to have a product spectrum inconsistent with that claim. It is proposed that this type II form does not differ sufficiently to be accepted as separate from type I. The third subtype, hereditary orotic aciduria without megaloblastic anaemia, occurs in two cases. It has the product spectrum expected of a defect in orotidine monophosphate decarboxylase. This form is the only one that appears to have a qualitatively different uridine monophosphate synthase. The possibility that orotidine monophosphate may control flux through the pyrimidine biosynthesis pathway in hereditary orotic aciduria is discussed. PMID- 19562505 TI - Long-term neurodevelopmental effects of early detection and treatment in a 6-year old patient with argininaemia diagnosed by newborn screening. AB - Newborn screening makes possible the early identification and treatment of asymptomatic ARG1-deficient patients; however, it is unknown whether early intervention prevents neurological insults. We identified a full-term Hispanic male infant with argininaemia by newborn screening with a serum arginine of 327 umol/L (reference values 0-140); ARG1 was undetectable on enzyme assay. Sequence analysis of ARG1 revealed a heterozygous nonsense mutation, c.223A>T (p.K75X), and a novel heterozygous missense variant, c.425G>A (p.G142E). Dietary protein restriction began from age 3 months, with addition of sodium benzoate at 4 months, and carnitine from 14 months. For the past 6 years, his serum arginine concentrations were maintained between 268 and 763 umol/L (reference values 10 140). He has normal development without spastic paraplegia, but with mild hepatomegaly and stable hepatic dysfunction. A full neurodevelopmental assessment was conducted at age 5 years. The BASC-2 rated the patient's behaviours as age appropriate. The Leiter-R assessed his 'Fundamental Visualization', 'Sequential Order', and 'Picture Concept' at 'Average', 'Form Completion' and 'Matching' at 'Low Average', and 'Figure Ground' and 'Repeated Patterns' in the 'Deficit' range. The full-scale IQ and the functioning ability presented in the 'Borderline' range and in the 'Low Average' range, respectively. The VABS/Survey Spanish Version showed difficulty in receptive and written language and fine and gross motor skills, and his performance to be at younger than his chronological age. The Short Sensory Profile showed some difficulty with taste and smell sensitivity. Long-term observation over 6 years in a patient with early treated argininaemia shows promising neurodevelopmental results. PMID- 19562504 TI - Sleep-related breathing in children with mucopolysaccharidosis. AB - BACKGROUND: The mucopolysaccharidoses (MPSs), a group of genetic lysosomal storage disorders, are associated with significant morbidity. Secondarily to specific associated anatomical abnormalities, MPS is associated with sleep disordered breathing (SDB), specifically obstructive sleep apnoea (OSA) that may confer additional morbidity. Few studies have examined SDB in children with MPS using full polysomnography (PSG) and thus the exact prevalence and severity of SDB is unknown. Further, successful treatments for SDB in this population have not been explored. OBJECTIVES: This study evaluated both SDB and the efficacy of treatments offered to children with MPS using PSG data. PATIENTS AND METHODS: A retrospective chart review was conducted on all children with MPS and a history of suspected OSA who were referred to the Hospital for Sick Children, Toronto. Both baseline and follow up treatment PSG data were analysed. PSG data recorded included obstructive apnoea-hypopnoea index (OAHI) and central apnoea index (CAI). RESULTS: Fourteen patients (10 male) underwent a baseline PSG. Three of 14 children on ERT were excluded from the main analyses. The median (range) baseline parameters of the population (n = 11) were recorded. The age was 5.2 years (0.8 17.8) and the body mass index (BMI) was 19.9 (13.7-22.2). The OAHI was 6.6 (0.0 54.8); the CAI was 0.6 (0.0-2.6). Seven of 11 (64%) had evidence for OSA and 3/7 children were classified as having severe OSA (OAHI > 10). Of these, 5/7 children underwent treatment for OSA with 3/5 children showing a significant reduction in their OAHI. Further, the 2 patients on ERT therapy with OSA were also both successfully treated. CONCLUSIONS: Children with MPS have a high prevalence of significant OSA and thus should be carefully screened for OSA using full polysomnography and treated accordingly. PMID- 19562506 TI - Primary cutaneous mucormycosis caused by Rhizomucor variabilis in an immunocompetent patient. AB - Primary cutaneous mucormycosis is an uncommon disease and occurs mainly in patients with immunocompromised disorders. We report a case of cutaneous mucormycosis in an immunocompetent man in whom no definite precipitating factors were noted. The isolate was identified as Rhizomucor variabilis according to the fungus morphology and DNA sequencing results. The lesion was successfully treated with oral Itraconazole, although the in vitro drug-susceptibility test showed resistance. PMID- 19562507 TI - Molecular cloning, sequence and expression analysis of ZmArf2, a maize ADP ribosylation factor. AB - A full-length cDNA encoding a maize GTP-binding protein of the ADP-ribosylation factor family was cloned by suppression subtractive hybridization and an in silico cloning approach. The cDNA was 938 bp in length and contained a complete ORF of 612 bp, which encodes a protein of 203 amino acid residues. Its deduced amino acids sequence had an 83% identity with that of a GTP-binding protein in rice. The gene was designated ZmArf2. The ZmArf2 gene consists of G1, G2, G3, G4 and G5 boxes, and Switch I and Switch II regions. Eight nucleotides differed and five amino acids changed between the popcorn inbred N04 and the dent corn inbred Dan232. One changed amino acid was in the G1 box. RT-PCR analysis showed that ZmArf2 expression increased in the early stages of endosperm development and was not tissue-specific. PMID- 19562508 TI - Expression of biologically active recombinant antifreeze protein His-MpAFP149 from the desert beetle (Microdera punctipennis dzungarica) in Escherichia coli. AB - An insect antifreeze protein gene Mpafp149 was cloned by the RT-PCR approach from the desert beetle Microdera punctipennis dzungarica. Sequence analysis revealed that this gene encoding a protein of 120 amid acids and this protein showed 65 76% homology with other insect antifreeze proteins, the deduced amino acid sequence displays very high similarities in those regions that contain tandem the 12-residue repeats (TCTxSxxCxxAx) domain and the TCT motif. Mpafp149 gene was cloned into pET-28a vector and expressed in Escherichia coli. A single-step purification based on specific binding of histidine residues was achieved. The purified His-MpAFP149 was SDS-PAGE analyzed, showing an atypical migration with molecular weight of about 24 kDa. The expression of His-MpAFP149 was confirmed by Western blot with specific binding to anti-GST-MpAFP149 antibody. The thermal hysteresis activity of the purified recombinant protein was 0.915 degrees C at 0.09 mg/ml, and the supercooling point was -9.6 degrees C at 0.03 mg/ml. In vitro antifreeze activity assay by measuring the survival rate of bacteria at -7 and 20 degrees C respectively, with the protection of His-MpAFP149 showed that the His-MpAFP149 fusion protein was able to enhance the freeze resistance of bacteria. PMID- 19562509 TI - Association of functional polymorphisms in MMPs genes with gastric cardia adenocarcinoma and esophageal squamous cell carcinoma in high incidence region of North China. AB - The aim of the present study was to investigate the association of single nucleotide polymorphisms (SNPs) in matrix metalloproteinase (MMPs) with the risk of gastric cardia adenocarcinoma (GCA) and esophageal squamous cell carcinoma (ESCC). Genotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism method in 592 patients and 624 healthy individuals. Significant differences in allele and genotype distributions of MMP-2 -1306C --> T SNP were observed between ESCC and controls (P = 0.02 and 0.01, respectively). Compared with the C/T + T/T genotypes, C/C genotype significantly increased the risk of ESCC (OR = 1.57, 95% CI = 1.10-2.23), especially in individuals in smoker group and in the group with positive family history. The stratification analysis showed there were risk changes of GCA for -735C/C genotype carrier in nonsmoker, for MMP-12 -82G allele and MMP-13 -77A/G genotype carrier in smoker. Our study indicated that these four functional polymorphisms might play roles in developing ESCC and GCA in high incidence region of North China. PMID- 19562510 TI - Pre and post cloning characterization of a beta-1,4-endoglucanase from Bacillus sp. AB - Consistent with its precloning characterization from the cellulolytic Bacillus sp., beta-1,4-endoglucanase purified from the recombinant E. coli exhibited maximum activity at 60 degrees C and pH 7.0. It was highly specific for CMC hydrolysis, with stability up to 70 degrees C and over a pH range of 6.0-8.0. The K(m) and V(max) values for CMCase activity of the enzyme were 4.1 mg/ml and 25 micromole/ml min(-1), respectively. The purified enzyme was a monomer of 65 kDa, as determined by SDS-PAGE. The presence of sucrose and IPTG in fermentation media increased the endoglucanase activity of the recombinant enzyme to 5.2-folds as compared with that of the actual one. PMID- 19562511 TI - Evaluation of a reverse-hybridization StripAssay for the detection of genetic polymorphisms leading to acenocoumarol sensitivity. AB - Acenocoumarol is mainly catabolized by CYP2C9 isoform of cytochrome P450 (CYP) liver complex and exerts its anticoagulant effect through the inhibition of Vitamin K Epoxide Reductase (VKOR). The most important genetic polymorphisms which lead to an impaired enzymatic activity and therefore predispose to acenocoumarol sensitivity, are considered to be CYP2C9*2 (Arg144Cys), CYP2C9*3 (Ile359Leu) and VKORC1-1639G>A, respectively. In this study we compared the results of the PGXThrombo StripAssay kit (ViennaLab Diagnostics,Vienna, Austria) with direct DNA sequencing and in house Restriction Fragment Length Polymorphisms (RFLP) for the detection of the aforementioned Single Nucleotide Polymorphisms (SNPs). The reverse hybridization StripAssay was found to be equally effective with RFLP and direct DNA sequencing for the detection of CYP2C9*2 and CYP2C9*3 polymorphisms, respectively. The comparison of the RFLP reference method with the reverse hybridization StripAssay for the detection of VKORC1-1639 G>A polymorphism showed that the reverse hybridization StripAsssay might misclassify some A/A homozygotes as heterozygotes. Optimization of the hybridization procedures may eliminate the extra low signal band observed in some samples at the reverse hybridization StripAssay and improve its diagnostic value. PMID- 19562512 TI - Isolation and heterologous transformation analysis of a pollen-specific promoter from wheat (Triticum aestivum L.). AB - The promoter of a pollen-specific gene TaPSG719 was isolated from wheat (Triticum aestivum L.) by inverse-PCR (IPCR). Sequence analysis revealed that the promoter contains two cis-acting elements (AGAAA and GTGA) known to confer anther/pollen specific gene expression which suggests that the promoter of TaPSG719 gene is a pollen-specific one. To ascertain the regulatory function of TaPSG719 promoter, two deleted fragments (-1,776 to -1 bp and -1,019 to -1 bp) were fused to the beta-glucuronidase (GUS) gene and transformed into tobacco plants. Similar GUS expression patterns were observed in all transformed plants and its activity was detected exclusively in pollen. No GUS activity in any other floral or vegetative tissue was observed. The results confirm that TaPSG719 promoter is pollen specific and active during the middle stages of pollen development till anther matured, and it can drive pollen-specific gene expression across the species. PMID- 19562513 TI - Psychosocial adaptation status and health-related quality of life among older Chinese adults with visual disorders. AB - PURPOSE: To examine the association of psychosocial adaptation status with vision specific health-related quality of life (HRQOL) and the role of psychosocial adaptation in the linkage between visual impairment and vision-specific HRQOL outcomes among older adults with visual disorders. DESIGN AND METHODS: In this cross-sectional study, older urban adults with visual problems (N = 167) were interviewed using a structured questionnaire to assess their self-reported visual function, general health, psychosocial adaptation status, and vision-specific HRQOL. Performance-based measure of visual function marked by distance visual acuity was clinically conducted by ophthalmologists. RESULTS: It was found in the study that psychosocial adaptation status was significantly associated with vision-specific HRQOL, including the domains of mental health symptoms due to vision and dependency on others due to vision. The results also showed that psychosocial adaptation status could buffer the effect of visual impairment on vision-specific HRQOL, including the domains of social function, mental health, and dependency. CONCLUSION: Psychosocial adaptation status is significantly associated with multiple domains of vision-specific HRQOL. The findings have significant implications for health education and psychosocial intervention for older adults with age-related vision loss. PMID- 19562515 TI - Quality of life in Serbian patients with Parkinson's disease. AB - BACKGROUND: The Parkinson's Disease Questionnaire (PDQ-39) is a well-validated British scale for the assessment of health-related quality of life (QoL) in Parkinson's disease (PD). OBJECTIVE: To validate the Serbian version of the PDQ 39, while also providing additional information on the characteristics of this instrument. PATIENT AND METHODS: A total of 102 Serbian PD patients were asked to complete the PDQ-39, a disease-specific QoL questionnaire, as well as the generic, health status questionnaire (SF-36-version 1), and the 21-item Beck Depression Inventory. Neurological examination included the Hoehn and Yahr staging, Unified Parkinson's disease rating scale (UPDRS)-part III, Schwab and England scale, and the Mini-Mental State Examination. RESULTS: Internal consistency analysis yielded a Cronbach's alpha of 0.83. Cronbach's alpha was above 0.70 for seven out of eight subscales (range from 0.73 to 0.91). A hierarchical structure of the PDQ-39 was revealed, with one global higher-order factor and two lower-order factors. The strongest predictor of the QoL in PD was the presence of depression, while motor disability (UPDRS-part III score) additionally contributed to poor QoL. Cognitive impairment has not been correlated with poor QoL. Also, QoL measures were not different between young- (< or =50 years) and older-onset PD patients. CONCLUSIONS: The PDQ-39 is a reliable and valid instrument for the assessment of QoL in Serbian PD patients. PMID- 19562514 TI - The concurrent validity and responsiveness of the health utilities index (HUI 3) among patients with advanced HIV/AIDS. AB - OBJECTIVES: To assess the concurrent validity and responsiveness of the Health Utility Index 3 (HUI3) in patients with advanced HIV/AIDS, and to determine the responsiveness of this measure, the MOS-HIV and EQ-5D to HIV-related clinical events. METHODS: Data from the OPTIMA (OPTions In Management with Antiretrovirals) trial was analyzed. Two aspects of the validity of the HUI3 were considered: concurrent validity was evaluated using Spearman correlations with MOS-HIV component and summary scores. Responsiveness to AIDS-defining events (ADE) and all adverse events (our external change criterion) was assessed using area under the receiver operating characteristic (AUROC) curves. RESULTS: The study enrolled 368 patients (mean follow-up: 3.66 years); 82% had at least one severe adverse event and 27% had at least one ADE. The HUI3 scale and items showed good concurrent validity, with 85% of the expected relationships with the MOS-HIV subscales verified. The HUI3 was responsive to both adverse events (AUROC [95%CI]: 0.68 [0.57, 0.80]) and ADEs (0.62 [0.51, 0.74]). The EQ-5D was responsive to ADEs (0.66 [0.56, 0.76]), but not responsive to adverse events (0.56 [0.46, 0.68]). CONCLUSION: The HUI3 is a valid and responsive measure of the change in HRQoL associated with clinical events in an advanced HIV/AIDS population. PMID- 19562516 TI - Infections after bone allograft surgery: a prospective study by a hospital bone bank using frozen femoral heads from living donors. AB - In the advent of the EU guidelines 2004/23/EG and 2006/17/EG requiring extensive safety and quality steps in bone banking, the prevalence and risk of infection disease transmission from bone allograft needs to be reconsidered. Therefore, we prospectively reviewed the screening process of bone donations and the outcome of surgeries utilizing bone allografts from our internal hospital bone bank with regard to infections according to CDC criteria. One-hundred and eighty-eight allogenic bone transplantation procedures in 160 patients were followed-up for 12 64 months (mean 32 months). Bacterial infection occurred in 11 patients, the overall infection rate therefore was 6.9%. After review of the clinical and intraoperative findings, none of the infections were likely to have been caused by the bone graft. Although no follow-up serologic testing was performed, no HIV of hepatitis infections were observed. Frozen bone allografts derived from live donors and provided by hospitals can generally be considered safe. However, without new and relevant clinical expertise, continuing this technique will be impeded by the new EU guidelines and their national implementations. PMID- 19562518 TI - Editors' overview: forbidding science? PMID- 19562517 TI - Distributed processing and temporal codes in neuronal networks. AB - The cerebral cortex presents itself as a distributed dynamical system with the characteristics of a small world network. The neuronal correlates of cognitive and executive processes often appear to consist of the coordinated activity of large assemblies of widely distributed neurons. These features require mechanisms for the selective routing of signals across densely interconnected networks, the flexible and context dependent binding of neuronal groups into functionally coherent assemblies and the task and attention dependent integration of subsystems. In order to implement these mechanisms, it is proposed that neuronal responses should convey two orthogonal messages in parallel. They should indicate (1) the presence of the feature to which they are tuned and (2) with which other neurons (specific target cells or members of a coherent assembly) they are communicating. The first message is encoded in the discharge frequency of the neurons (rate code) and it is proposed that the second message is contained in the precise timing relationships between individual spikes of distributed neurons (temporal code). It is further proposed that these precise timing relations are established either by the timing of external events (stimulus locking) or by internal timing mechanisms. The latter are assumed to consist of an oscillatory modulation of neuronal responses in different frequency bands that cover a broad frequency range from <2 Hz (delta) to >40 Hz (gamma) and ripples. These oscillations limit the communication of cells to short temporal windows whereby the duration of these windows decreases with oscillation frequency. Thus, by varying the phase relationship between oscillating groups, networks of functionally cooperating neurons can be flexibly configurated within hard wired networks. Moreover, by synchronizing the spikes emitted by neuronal populations, the saliency of their responses can be enhanced due to the coincidence sensitivity of receiving neurons in very much the same way as can be achieved by increasing the discharge rate. Experimental evidence will be reviewed in support of the coexistence of rate and temporal codes. Evidence will also be provided that disturbances of temporal coding mechanisms are likely to be one of the pathophysiological mechanisms in schizophrenia. PMID- 19562519 TI - Abstracts of the 7th European Cytogenetics Conference. July, 4-7, 2009. Stockholm, Sweden. PMID- 19562522 TI - A closed concept of extractive whole cell microbial transformation of benzaldehyde into L-phenylacetylcarbinol by Saccharomyces cerevisiae in novel polyethylene-glycol-induced cloud-point system. AB - Extractive microbial transformation of benzaldehyde into L-phenylacetylcarbinol (L-PAC) by Saccharomyces cerevisiae (Baker's yeast) has been carried out in a novel polyethylene-glycol-induced cloud-point system (PEG-CPS). The extractive microbial transformation in the PEG-CPS and a downstream process for stripping of the product from the microbial transformation broth with microemulsion extraction are demonstrated. The results indicate that the PEG-CPS maintains the advantage of CPS for in situ extraction of polar product in the microbial transformation. At the same time, the utilization of hydrophilic nonionic surfactant in the PEG CPS is favorable for stripping of product from the nonionic surfactant in the microbial transformation broth by Winsor I microemulsion extraction. Thus, a closed concept of in situ extraction of polar product in microbial transformation and its downstream process of product recovery are fulfilled at the same time. PMID- 19562523 TI - Treatment of sanitary landfill leachates in a lab-scale gradual concentric chamber (GCC) reactor. AB - Sanitary landfill leachates are a major environmental problem in South American countries where sanitary landfills are still constructed and appropriate designs for the treatment of these leachates remain problematic. The performance of a lab scale Gradual Concentric Chamber (GCC) reactor for leachates treatment is presented in this study. Two types of sanitary landfill residuals were evaluated, one directly collected from the garbage trucks (JGL), with high organic strength (84 g COD/l) and the second one, a 6-month-generated leachate (YL) collected from the lagoon of the sanitary landfill in Quito, Ecuador, with an organic strength of 66 g COD/l. Different operational parameters, such as organic loading rate (OLR), temperature, recycling and aeration, were tested. The GCC reactor was found to be a robust technology to treat these high-strength streams with organic matter removal efficiencies higher than 65%. The best performance of the reactors (COD removal efficiencies of 75-80%) was obtained at a Hydraulic Retention Time (HRT) of around 20 h and at 35 degrees C, with an applied OLR up to 70 and 100 g COD/l per day. Overall, the GCC reactor concept appears worth to be further developed for the treatment of leachates in low-income countries. PMID- 19562524 TI - Antiphospholipid antibody-mediated reproductive failure in antiphospholipid syndrome. AB - The association of elevated titers of circulating antiphospholipid (anti-PL) Abs in antiphospholipid syndrome (APS) and reproductive failure is well established in the literature. The clinical features include recurrent abortions at various stages, including implantation, placentation in the first trimester, miscarriages in the second and third trimesters, intrauterine growth retardation, preeclampsia with placental insufficiency and growth restrictions, arterial and venous thrombosis, and possibly also infertility. APS-mediated recurrent pregnancy loss and other features of reproductive failure might result from diverse autoimmune factors, inflammation, involving different mechanisms, which encompass pathogenic anti-PL Abs. Herein, we discuss the association of anti-PL Abs with reproductive failure with special emphasis on antiphospholipid autoantibodies characterizing APS. This association is evident from either human studies or murine models. PMID- 19562525 TI - Amyloid-beta induces a caspase-mediated cleavage of P2X4 to promote purinotoxicity. AB - Overproduction of the beta-amyloid fragment 1-42 (A beta(1-42)) is thought to contribute to synaptic dysfunction and neuronal death in Alzheimer's disease. Mounting evidence suggests that purinergic receptors play critical roles in synaptic plasticity and neuronal survival, but the potential involvement of these receptors in A beta(1-42)-induced synaptic dysfunction and neuronal death has not been addressed. Here we report that A beta(1-42) promoted accumulation of the calcium-permeable purinergic receptor P2X4 in neurons. We also report evidence that A beta(1-42) induced a caspase-3-mediated cleavage of the receptor that slowed channel closure times and prevented agonist-induced internalization of the receptor. Molecular interference to reduce the expression of P2X4 in primary rodent neurons attenuated A beta(1-42)-induced neuronal death while induced expression of P2X4 in a neuronal cell line that does not normally express P2 receptors enhanced the toxic effect of A beta(1-42). Together these findings suggest that A beta(1-42)-induced synaptic dysfunction and neuronal death may involve perturbations in P2X4 purinergic receptors. PMID- 19562526 TI - Nucleic acids electrotransfer-based gene therapy (electrogenetherapy): past, current, and future. AB - About 25 years after the publication of the first report on gene transfer in vitro in cultured cells by the means of electric pulses delivery, reversible cell electroporation for gene transfer and gene therapy (DNA electrotransfer) is at a cross in its development. Present knowledge on the effects of cell exposure to appropriate electric field pulses, particularly at the level of the cell membrane, is reported here. The importance of the models of electric field distribution in tissues and of the correct choice of electrodes and applied voltages is highlighted. The mechanisms involved in DNA electrotransfer, which include cell electropermeabilization and DNA electrophoresis, are also surveyed. This knowledge has allowed developing new nucleic acids electrotransfer conditions using combinations of permeabilizing pulses of high voltage and short duration, and of electrophoretic pulses of low voltage and long duration, which are very efficient and safer. Feasibility of electric pulses delivery for gene transfer in humans is discussed taking into account that electric pulses delivery is already regularly used for localized drug delivery in the treatment of cutaneous and subcutaneous solid tumors by electrochemotherapy. Because recent technological developments made DNA electrotransfer more and more efficient and safer, this non-viral gene therapy approach is now ready to reach the clinical stage. A good understanding of DNA electrotransfer principles and the respect of safe procedures will be key elements for a successful future transfer DNA electrotransfer into the clinics. PMID- 19562528 TI - Epidemiology of atrial fibrillation. AB - Atrial fibrillation is the most common sustained cardiac arrhythmia in the general population. Unfortunately, current treatment strategies aiming at the elimination of atrial fibrillation have limited long term success rates and significant risks. In this context, recent publications have provided many insights on potentially treatable risk factors for the occurrence of atrial fibrillation, such as alcohol, blood pressure, obesity, inflammation and nutritional factors. In this review, we summarise the current evidence on these risk factors and indicate areas in need of further investigation. The current evidence shows that blood pressure, hypertension and obesity seem to play a key role in the pathogenesis of atrial fibrillation. Preliminary evidence also suggests that inflammation is an important mediator of these associations. Knowledge of these interrelationships may eventually help to develop new treatment strategies and decrease the burden of atrial fibrillation in the general population. PMID- 19562530 TI - Oscillometric measurement of ankle-brachial index in patients with suspected peripheral disease: comparison with Doppler method. AB - QUESTION UNDER STUDY: Purpose was to validate accuracy and reliability of automated oscillometric ankle-brachial (ABI) measurement prospectively against the current gold standard of Doppler-assisted ABI determination. METHODS: Oscillometric ABI was measured in 50 consecutive patients with peripheral arterial disease (n = 100 limbs, mean age 65 +/- 6 years, 31 men, 19 diabetics) after both high and low ABI had been determined conventionally by Doppler under standardised conditions. Correlation was assessed by linear regression and Pearson product moment correlation. Degree of inter-modality agreement was quantified by use of Bland and Altman method. RESULTS: Oscillometry was performed significantly faster than Doppler-assisted ABI (3.9 +/- 1.3 vs 11.4 +/- 3.8 minutes, P <0.001). Mean readings were 0.62 +/- 0.25, 0.70 +/- 0.22 and 0.63 +/- 0.39 for low, high and oscillometric ABI, respectively. Correlation between oscillometry and Doppler ABI was good overall (r = 0.76 for both low and high ABI) and excellent in oligo-symptomatic, non-diabetic patients (r = 0.81; 0.07 +/ 0.23); it was, however, limited in diabetic patients and in patients with critical limb ischaemia. In general, oscillometric ABI readings were slightly higher (+0.06), but linear regression analysis showed that correlation was sustained over the whole range of measurements. CONCLUSIONS: Results of automated oscillometric ABI determination correlated well with Doppler-assisted measurements and could be obtained in shorter time. Agreement was particularly high in oligo-symptomatic non-diabetic patients. PMID- 19562529 TI - Management of transitional-cell carcinoma of the renal pelvis and ureter. AB - Transitional-cell carcinoma of the renal pelvis or ureter is a relatively rare disease. Several risk factors are smoking, occupational carcinogens, analgesic abuse or Balkan nephropathy. The grade and stage of the disease have the most significant impact on the outcome. The treatment of renal pelvis and ureter tumours is open or laparoscopic surgery varying from conservative to more extensive surgical procedures, i.e. radical nephroureterectomy including removal of the contents of Gerota's fascia with ipsilateral ureter and a cuff of bladder at its distal extent. Most available data are from retrospective studies and surgery is the mainstay of treatment. Chemotherapy and/or radiation therapy are possible adjuvant or primary treatment for selected patients; however, prospective studies are needed to confirm their use. PMID- 19562532 TI - [An integrated psychosocial-spiritual model for cancer pain management]. AB - Cancer pain is known to be a multidimensional and complex experience that can cause severe suffering and can lessen the quality of life. The psychosocial and spiritual aspects of cancer pain play an important role in this phenomenon. This article describes a multidisciplinary model for cancer pain management that focuses on the psychosocial and spiritual aspects of cancer pain and the needs of patients. PMID- 19562531 TI - Sperm mediated human coagulation factor VIII gene transfer and expression in transgenic mice. AB - PRINCIPLES: B-domain deleted human coagulation factor VIII cDNA (BDD-hFVIIIcDNA) transgenic mice were produced by using sperm mediated gene transfer (SMGT). The transcription and expression of human FVIII in transgenic mice were also investigated. METHODS: Sperm were isolated from caudae epididymides of male C57BL/6 mice and transfected with linearized RC/RSV-BDDhFVIIIcDNA plasmid, and subsequently used to fertilize female mice via artificial insemination in vivo. After birth, F0 progeny were identified by PCR and Southern blotting for BDD hFVIIIcDNA transgenic mice. F1 progeny were subsequently derived from a male transgenic F0 mouse and a normal C57BL/6 female mouse. The F1 progeny were then identified as BDD-hFVIIIcDNA transgenic mice by Southern blotting. The transcription and expression of BDDhFVIIIcDNA in transgenic mice were determined by northern blotting, western blotting and immunohistochemical staining. Blood was also collected from both F0 and F1 progeny to detect hFVIII:Ag and anti hFVIII inhibitors. RESULTS: A total of 9 F0 and 8 F1 progeny were delivered, in which 3 F0 and 2 F1 progeny were identified to have BDD-hFVIIIcDNA. The transcription and expression of BDD-hFVIIIcDNA were found to exist in the liver and kidneys of all transgenic mice. HFVIII:Ag in plasma of the transgenic F0 progeny was 31.95 ng/ml, 23.52 ng/ml and 26.36 ng/ml respectively, whilst the F1 transgenic mice showed results of 18.82 ng/ml and 12.16 ng/ml. Anti-hFVIII inhibitors were negative in both F0 and F1 progeny. CONCLUSIONS: Human FVIII gene transgenic mice can be produced by the SMGT technique and express human FVIII protein in their bodies. PMID- 19562527 TI - Role of membrane potential in the regulation of cell proliferation and differentiation. AB - Biophysical signaling, an integral regulator of long-term cell behavior in both excitable and non-excitable cell types, offers enormous potential for modulation of important cell functions. Of particular interest to current regenerative medicine efforts, we review several examples that support the functional role of transmembrane potential (V(mem)) in the regulation of proliferation and differentiation. Interestingly, distinct V(mem) controls are found in many cancer cell and precursor cell systems, which are known for their proliferative and differentiation capacities, respectively. Collectively, the data demonstrate that bioelectric properties can serve as markers for cell characterization and can control cell mitotic activity, cell cycle progression, and differentiation. The ability to control cell functions by modulating bioelectric properties such as V(mem) would be an invaluable tool for directing stem cell behavior toward therapeutic goals. Biophysical properties of stem cells have only recently begun to be studied and are thus in need of further characterization. Understanding the molecular and mechanistic basis of biophysical regulation will point the way toward novel ways to rationally direct cell functions, allowing us to capitalize upon the potential of biophysical signaling for regenerative medicine and tissue engineering. PMID- 19562533 TI - Preemptive analgesic effects of intravenous paracetamol in total abdominal hysterectomy. AB - OBJECTIVES: Paracetamol is primarily thought to be a cyclooxygenase inhibitor acting through the central nervous system. Indirect effects of paracetamol are through the serotoninergic system as a non-opioid analgesic. In this study, total abdominal hysterectomy patients were given intravenous (iv) paracetamol 1 g preoperatively or intraoperatively to assess its postoperative analgesic effects. METHODS: 90 patients undergoing total abdominal hysterectomy were enrolled into the study. Patients were randomized into three groups: in Group I, iv paracetamol 1 g was given 30 minutes prior to induction. In Group II, iv paracetamol 1 g was given prior to skin closure. Group III served as the control group and received saline as placebo. Postoperatively, all patients received morphine via patient controlled analgesia pump. Postoperatively, rest and activity pain scores, sedation scores, hemodynamic parameters, postoperative morphine consumption, side effects, patient satisfaction, and total hospital stay were recorded. RESULTS: In the control group, at rest and movement pain scores and total morphine consumption via patient-controlled analgesia were higher than in Groups I and II. When Groups I and II were compared, total morphine consumption was much greater in Group II. Intravenous paracetamol intraoperatively and postoperatively did not result in any hemodynamic effects. CONCLUSION: In total abdominal hysterectomy, preemptive iv paracetamol 1 g provided good quality postoperative analgesia, with decreased consumption of morphine and minimal side effects. PMID- 19562534 TI - [The effects of lumbar plexus block and epidural block on total blood loss and postoperative analgesia in total hip arthroplasty]. AB - OBJECTIVES: In this study, the effects of lumbar plexus block and epidural block on total blood loss and postoperative analgesia in patients undergoing total hip arthroplasty (THA) under general anesthesia was documented. METHODS: The study included 45 patients undergoing THA under general anesthesia. Group GA (n=15) received general anesthesia, Group GA+E (n=15) received general anesthesia + epidural catheter and Group GA+LPB (n=15) received general anesthesia + lumbar plexus catheterization. Intraoperative blood transfusion (IOBT) requirements and intraoperative bleeding (IOB) were documented. Postoperative hemoglobin, hematocrit and total blood loss through the drains were also documented. All patients received patient-controlled analgesia through the IV route. Time to first analgesic requirement (TFAR), total IV morphine consumption (MORPH 24) and 24-hour visual analogue scale (VAS) values were evaluated. RESULTS: TFAR was the lowest (8.7+/-4.0 min.) in the GA group and highest (42.7+/-14.4 min) in the GA+LPB group, and the GA+E group had lower values than the GA+LPB (32.3+/-16.0 min) group (p<0.05). IOBT, IOB, and MORPH 24 average values were the lowest in the GA+LPB group and highest in the GA group, with GA+E in between (p<0.05). Postoperative first VAS values were significantly different between the GA group and the others (p<0.001). There was also a significant difference between the GA group and the others regarding postoperative average Hb values (p<0.02). CONCLUSION: THA using either regional technique provides less blood loss and better analgesia. We will consider LPB in our future cases in view of the lower intraoperative blood loss and better analgesia. PMID- 19562535 TI - [The effects of two different multimodal analgesic regimens in total hip replacement surgery]. AB - OBJECTIVES: This study evaluated the effectiveness of two different multimodal analgesia protocols in terms of postoperative pain relief, tramadol consumption during patient-controlled analgesia (PCA) and side effects after total hip replacement surgery. METHODS: Group F (n=18) received general anesthesia after a 3-in-1 femoral nerve block (FNB) was applied using 40 ml of bupivacaine 0.25%. Group FD (n=18) received general anesthesia after the same block and dexketoprofen p.o. was given. All patients received intravenous tramadol at the end of surgery via a PCA device. Group FD was given dexketoprofen 75 mg/day for 48 hours postoperatively. Pain scores were evaluated at 0, 1/2, 1, 4, 8, 12, 24 and 48h at rest and on movement of the hip. Side effects and global satisfaction scores in both groups were also evaluated in this setting. RESULTS: Total tramadol consumption was lower in Group FD (377.7+/-137.4) than in Group F (593.9+/-132.3) (p<0.05). Visual analogue scale (VAS) scores were 0.05). Similarly, there were no significant differences between the incision groups according to upper or lower abdominal wall site of incision (p>0.05). CONCLUSION: Midline abdominal wall incisions of different lengths and placements (upper or lower abdomen) have similar dehiscence pressures against the pneumoperitoneum in the early postoperative period (p>0.05). PMID- 19562540 TI - [Evaluation of the process and effectiveness of consultation system in the Department of Emergency Medicine]. AB - BACKGROUND: Triage, consultations, and radiological and laboratory test processes have different effects on the total waiting time in the emergency department (ED). Under these circumstances, the importance of the consultation system process and effectiveness of consultation becomes very clear. Our aim in this study was to verify the process of the consultation system. METHODS: This prospective and defining study was performed with 276 patients admitted to the ED. A total of 342 consultations were requested. These patients were classified as very urgent, urgent and non-urgent according to their problems, and a survey form was completed by the ED resident. RESULTS: The most frequently requested consultation was to the Department of Internal Medicine (72%). Mean time for reply to the consultation was 29+/-43 minutes. The earliest reply to the consultation was from Cardiology while the latest responders were the general surgeons. Timeline for replying to the consultation was shorter depending on the urgency of the case. CONCLUSION: In our study, we determined that the most important factor for the effectiveness of consultation was the definition of the urgency of the patients by the residents in the ED. Since the number of patients admitting to the ED will continue to increase in the future, more detailed prospective studies are needed about the efficiency of consultation in the ED. PMID- 19562541 TI - [Factors affecting mortality in acute mesenteric obstruction]. AB - BACKGROUND: Acute mesenteric obstruction (AMO) is usually fatal. This study was designed to demonstrate the demographic characteristics and prognostic factors of affected patients. METHODS: The patients admitted to our emergency department and diagnosed as having AMO between January 2000 and December 2004 were investigated retrospectively. Their demographic characteristics, laboratory results, per operative findings and mortality were investigated retrospectively. RESULTS: Thirty patients (18 males, 12 females; mean age: 67 [26-92]) were evaluated. 43.3% of patients survived. Surviving patients had statistically significantly lower alanine aminotransferase (ALT) but also higher pH and creatinine levels (p=0.0027, 0.0004, 0.02). Colonic involvement also increased mortality (p<0.001). Papaverine infusion, embolectomy and second-look operations had no effect on outcome. CONCLUSION: Preoperatively increased liver enzymes, acidosis, and colonic involvement indicated poor prognosis. Papaverine infusion, embolectomy and second-look operations showed no advantages with respect to survival. PMID- 19562542 TI - [Esophageal foreign bodies: analysis of 188 cases]. AB - BACKGROUND: Esophageal foreign bodies (EFBs) represent an urgent clinical condition that can be seen in all ages, especially in children, and sometimes cause important morbidity and mortality. Rigid endoscopy is the most important diagnostic and treatment tool in EFBs, although there are some risks. METHODS: Between 1996-2006, the records of 188 inpatient cases (111 males [59%], 77 females [41%]; mean age 19+/-22.63; range 4 months to 96 years) who underwent rigid endoscopy were evaluated retrospectively. RESULTS: There was a history of foreign body in 158 cases (84%), and foreign body was shown by chest X-ray in 145 cases (77.1%). Of the foreign bodies, 137 (79.2%) were located in the hypopharynx and cervical esophagus. While the most commonly detected foreign bodies were coins (72 cases) and bones (42 cases), no foreign body was detected in 15 cases (8%) in rigid esophagoscopy. Foreign body was removed via surgery in 5 cases. Esophageal rupture as a complication during esophagoscopy occurred in 2 cases (1.06%). Mean hospital stay was 1.82+/-1.60 days. Mortality occurred in 1 case (.53%). CONCLUSION: Complications may be prevented with early diagnosis and accurate treatment. Rigid endoscopy is an effective and safe procedure for foreign body removal. PMID- 19562543 TI - [Traumatic extracranial carotico-juguler fistulation]. AB - BACKGROUND: Extracranial caroticovenous fistulae, if left untreated, may cause stroke, cerebral edema, and high output cardiac failure and may present with oculofacial signs. In this study, 5 cases with extracranial arteriovenous fistulae who were diagnosed and surgically treated promptly after trauma are presented. METHODS: Five patients with extracranial traumatic caroti-cojuguler fistulation underwent urgent surgery in our department. Their mean age was 32.4 (range: 27-38). Mean door to operating room time was 2.4 hours. One patient underwent arcus aortography due to a large hematoma in the mediastinum. One patient underwent arterial Doppler ultrasound examination. The remaining three patients were diagnosed via physical examination. RESULTS: There were three gunshot and two stab wounds. The right common carotid artery was injured in three cases and left common carotid artery in two. There was no early or late mortality. Cross-clamp time was 14.3+/-4.7 minutes. There was no major bleeding. One patient developed ischemic stroke secondary to hypotension due to massive bleeding before surgery and was transferred to the Department of Neurology on the 5th postoperative day. CONCLUSION: We suggest that as soon as the diagnosis of traumatic caroti-cojuguler fistula is made, a surgical approach is effective and safe and may prevent possible complications due to delayed diagnosis and treatment. PMID- 19562544 TI - Independent risk factors of morbidity in penetrating colon injuries. AB - BACKGROUND: The present study explored the factors effective on colon-related morbidity in patients with penetrating injury of the colon. METHODS: The medical records of 196 patients were reviewed for variables including age, gender, factor of trauma, time between injury and operation, shock, duration of operation, Penetrating Abdominal Trauma Index (PATI), Injury Severity Score (ISS), site of colon injury, Colon Injury Score, fecal contamination, number of associated intra and extraabdominal organ injuries, units of transfused blood within the first 24 hours, and type of surgery. In order to determine the independent risk factors, multivariate logistic regression analysis was performed. RESULTS: Gunshot wounds, interval between injury and operation > or =6 hours, shock, duration of the operation > or =6 hours, PATI > or =25, ISS > or =20, Colon Injury Score > or = grade 3, major fecal contamination, number of associated intraabdominal organ injuries >2, number of associated extraabdominal organ injuries >2, multiple blood transfusions, and diversion were significantly associated with morbidity. Multivariate logistic regression analysis showed diversion and transfusion of > or =4 units in the first 24 hours as independent risk factors affecting colon related morbidity. CONCLUSION: Diversion and transfusion of > or =4 units in the first 24 hours were determined to be independent risk factors for colon-related morbidity. PMID- 19562545 TI - Dangers faced by emergency staff: experience in urban centers in southern Turkey. AB - BACKGROUND: The aim of this study was to investigate the incidence and characteristics of aggression, threat and physical violence directed towards the staff in emergency departments. METHODS: A questionnaire was completed by the emergency staff. The individualized data collected included the pattern and incidence of violence, sex, age, profession, and years of experience of the emergency staff, and the behavioral characteristics of the assailants, together with outcome of incidents. Data regarding incidences occurring between May 1-31, 2006 were abstracted. RESULTS: A total of 109 staff were evaluated. There was a statistically significant relationship between aggression and profession (p=0.000), but no relation was determined with sex, age or years of experience (p values 0.464, 0.692, and 0.298, respectively). The relationship of incidences of threat with sex, age, profession, and experience was insignificant (p values 0.311, 0.278, 0.326, 0.994, respectively). On the other hand, significant relationships were identified between physical assault and sex, age, profession, and experience (p values 0.042, 0.000, 0.000, 0.011). CONCLUSION: Violence directed towards the emergency staff is common. Aggression occurs towards the emergency physician distinctively. Otherwise, there is no significant relationship between aggression or threat and personal characteristics. However, male sex, > or =31 years of age, being an emergency physician, and having worked for longer than five years in the emergency department are the risk factors for physical violence. PMID- 19562546 TI - How should open tibia fractures be treated? A retrospective comparative study between intramedullary nailing and biologic plating. AB - BACKGROUND: In this retrospective study, our purpose was to compare two treatment alternatives clinically. METHODS: Forty-five patients who had grade I or II open tibia fractures were included. Twenty-five of them, treated via minimally invasive plate osteosynthesis (MIPO), comprised group I. The latter 20 cases, treated via partial reamed intramedullary nailing (PR-IMN), comprised group II. Clinical evaluation was made on the basis of modified Ketenjian's criteria. RESULTS: Full weight-bearing periods in groups I and II were 21 and 22.4 weeks, respectively. Non-union in one case of group I was revised with circular fixator. In another case, implant removal was needed due to chronic osteomyelitis. Mal union was detected in another. In group II, two cases needed implant revision with intramedullary nail in one and circular fixator in another for non-union. Mal-union in one case and chronic osteomyelitis in another were the late complications in group II. At the last follow-up, satisfaction rates were: 21/25 in group I and 18/20 in group II. There was no significant difference between groups (p>0.05). CONCLUSION: The clinical results of both groups were similar. Although intramedullary nailing is the first choice, MIPO is an alternative method for open tibia fractures. PMID- 19562547 TI - Open reduction and internal fixation of radial head fractures. AB - BACKGROUND: We evaluated the radiologic and functional outcomes of patients with radial head fractures managed with open reduction and internal fixation. METHODS: Between 1998-2003, 15 patients (7 males, 8 females; mean age 34.1; range 18 to 49 years) with radial head fracture were treated with open reduction and internal fixation. Follow-up time was 54.6 months (42-78). Three fractures were Mason type II, 8 were III and 4 were IV. They were evaluated by anteroposterior and lateral radiographs and functionally by Broberg and Morrey criteria. RESULTS: All the fractures except in 1 patient with Mason type III had united. The mean range of motion of the elbow was 20 degrees to 145 degrees with 71.9 degrees of pronation and 83.2 degrees of supination. According to Broberg and Morrey criteria, the outcome was excellent in 8, good in 4, fair in 1 and bad in 2. Excision and prosthetic replacement were performed in 1 patient because of implant failure. CONCLUSION: We suggest open reduction and internal fixation even in comminuted cases because it gives satisfactory elbow function and avoids radial shortening, loss of motion and wrist joint dysfunction as a result of radial head excision. When it fails, excision and prosthetic replacement can be done later. PMID- 19562548 TI - Leg length discrepancies in adult femoral shaft fractures treated with intramedullary nailing. AB - BACKGROUND: To evaluate the leg length discrepancy (LLD) retrospectively in adult femoral shaft fractures treated with intramedullary nailing (IMN). METHODS: Sixty three patients (58 male, 5 female; mean age 29.9+/-12.4; range 15 to 77 years) were included in the study. Fractures were identified according to the Winquist Hansen (W) system and AO classification. 16 W0, 18 WI, 16 WII, 7 WIII, and 6 WIV fractures and 35 type A, 22 type B, and 6 type C fractures were repaired. Thirty one (49.2%) patients had multiple injuries. Fourteen patients sustained an open fracture. LLDs were measured on physical examination and using orthoroentgenography. RESULTS: The mean follow-up was 90.2+/-29.9 (39-193) months. The mean LLD was 12.3+/-15.2 [12-(-60)] mm using orthoroentgenography and 12.9+/-13.7 [10-(-60)] mm according to manual measurement. In seven cases, no LLD was observed. Twenty-seven shortenings and one lengthening were observed in the 28 femurs with a discrepancy greater than 10 mm (44.4%). There was no statistical correlation between LLD and open or closed fracture (r=0.02, p=0.86), polytrauma (r=-0.09, p=0.47), or delayed surgery (p=0.31), but there was a tendency to a greater discrepancy in comminuted fractures (WIII, IV) (r=0.33, p=0.007). CONCLUSION: LLD may be seen in high rates in adult femoral shaft fracture cases treated with IMN. Static IMN following absolute restoration of the length may prevent this problem in femoral diaphysis fractures, especially comminuted WIII and IV types. PMID- 19562549 TI - A retrospective study on the epidemiology and treatment of maxillofacial fractures. AB - BACKGROUND: Maxillofacial injuries constitute a substantial proportion of cases of trauma. This descriptive analytical study assesses the cause, type, incidence, and demographic and treatment data of maxillofacial fractures. METHODS: A retrospective study on maxillofacial traumas was carried out in the Department of Plastic and Reconstructive Surgery at Sisli Etfal Hospital (Istanbul, Turkey) between January 1, 2000 and December 31, 2005. The study included 216 patients with a mean age of 29.8 years. Sex and age distribution of patients, etiology of trauma, localization of the fractures, treatment modalities, time to treatment after the trauma, and postoperative complications were recorded. RESULTS: The male predilection was 75.5%. Road traffic accident was the most common causative factor (67.1%), followed by interpersonal violence (19.4%), falls (12.5%), and work- and sport-related accidents (0.9%). A total of 50% of the patients suffered isolated mandibular fractures, 23.6% had isolated midface fractures, and 26.3% had combined midface and mandibular fractures. Regarding distribution of mandibular fractures, the majority (26.8%) occurred in the parasymphysis, 14.8% in the angulus, and 11.1% each in the symphysis and corpus. Complications occurred in 6% of patients, and the most common was malocclusion followed by infection and nonunion. CONCLUSION: The causes and pattern of maxillofacial fractures reflect trauma patterns within the community and, as such, can provide a guide for the design of programs geared toward prevention and treatment. PMID- 19562550 TI - Treatment of type I capitellar fractures in adolescents. AB - BACKGROUND: Because fractures of the capitellum are rare in childhood and the young adolescent period, their treatment is still debatable and there appears to be no established treatment protocol. In the present study, we evaluated the results obtained in adolescents with type 1 capitellar fractures who were treated with open reduction and internal fixation with a 3.5 mm lag screw, directed from posterior to anterior. METHODS: Twelve type I capitellar fractures in adolescents were treated with open reduction and internal fixation with a single 3.5 mm cortical lag screw directed from the posterior to the anterior and the results were evaluated by an objective evaluation score (Broberg and Morrey's functional rating index). Mean age of the patients was 13.5. RESULTS: At final examination (24 to 90 months follow-up), mean Broberg and Morrey's functional rating index was 96.7 points (91 to 100 points). All fractures had healed in anatomic position and no avascular necrosis or heterotrophic ossification was observed. CONCLUSION: Since it is essential to obtain the full range of motion at the elbow, accurate open reduction and stable internal fixation are best to manage displaced type I capitellar fracture in children and adolescents. Single cortical lag screw directed from posterior to the anterior without penetrating the joint surface is suitable for this purpose. PMID- 19562552 TI - [Falls from height in childhood in Diyarbakir province: a questionnaire study combined with clinical data]. AB - BACKGROUND: In Diyarbakir, the rate of falls from height increases during summer months since people sleep on the unprotected balconies and roofs of their houses. We aimed to determine the frequency of falls from height and the related risk factors. METHODS: We used questionnaire method and clinical data in this study. The questionnaire was administered in 1445 residences. Clinical data were obtained from files of patients hospitalized due to trauma in the Neurosurgery Clinic of Dicle University over the last six years. RESULTS: In this questionnaire study, it was determined that 246 people had fallen, their average age was 15.4 years, 98% of them had fallen accidentally (mostly from 3.6 m height in summer months), mortality was 6.9%, and the prevalence of falls from height over the previous six years was 472/100,000. Of 464 patients who were hospitalized during the last six years, 326 were cases of fall from height and 59% of these patients were falls from a roof. The average age of these patients was 8.9 years, and average height of the fall was 4.2 m. CONCLUSION: Falls from height, particularly from roofs in Diyarbakir, remain a serious problem in terms of public health. PMID- 19562551 TI - Prognostic factors in severely traumatized eyes with posterior segment involvement. AB - BACKGROUND: To determine the prognostic indicators of functional outcome in eyes with severe posterior segment trauma managed with pars plana vitrectomy. METHODS: One hundred and six eyes of 101 patients were retrospectively reviewed to determine the accuracy of a number of factors in predicting functional outcome after surgery. These potential prognostic indicators included initial visual acuity (VA), retinal detachment (RD), type of trauma, presence of intraocular foreign body (IOFB), type of IOFB, posttraumatic endophthalmitis, hyphema, choroidal detachment, initial hypotonia, accompanying lens subluxation/dislocation, and severe vitreous hemorrhage. In our study, functional success was defined as VA > or =5/200. Fisher's exact and chi-square tests were used for statistical analysis. RESULTS: The mean follow-up time was 12.8+/-0.52 (8-18) months. Thirty-three eyes (31.13%) had functional success. Forty-four (68.7%) of 64 eyes with preoperative RD had anatomical success (total retinal reattachment). Predictors of poor visual outcome (VA < or =5/200) were found to be poor initial VA (p<0.0001), presence of RD (p<0.001), and presence of endophthalmitis (p<0.05). No statistically significant correlation was found between the other predictors surveyed and visual outcome (p>0.05). CONCLUSION: Vitreoretinal surgery can improve anatomical and functional success in eyes with severe posterior segment trauma. Poor initial VA, RD, and posttraumatic endophthalmitis are poor predictors of visual outcome. PMID- 19562553 TI - [Role of anatomic and physiologic trauma scoring systems in forensic cases]. AB - BACKGROUND: The Turkish Penalty Law has recently been changed. The novel law asks the practitioner to report if there is any 'Situation Placing a Life in Danger '. Herein, we evaluate the anatomic (ISS, TRISS) and physiologic (RTS) trauma scores assessing mortality. METHODS: Study data were obtained from a retrospective chart screening of cases who were accepted to the emergency department in GATA Faculty of Medicine in 2007 and from archived forensic reports. Demographic features and the time period of admittance were recorded. Trauma scores were calculated. All parameters were evaluated with the reported condition of "life threat" and mortality. RESULTS: Forensic reports were completed for 373 patients and 6.16% of them were noted as being in a life-threatening condition. Mortality rate was 1.34%. A significant rate of trauma patients suffered from firearm injury and stab wounds (p<0.001). There was no statistical difference between ISS, TRISS and RTS with respect to predictive value of a 'life-threatening condition' (Area under curve [AUC] in the receiver operating characteristic [ROC] curve analysis: ISS: 0.968, TRISS: 0.922, RTS: 0.196). There was also no statistical difference between ISS, TRISS and RTS scores regarding mortality prediction (AUC in the ROC analysis: ISS: 0.992, TRISS: 0.0995, RTS: 0.005). CONCLUSION: We assume that there is no difference between physiologic and anatomic scoring systems to predict mortality for deciding a life-threatening condition. PMID- 19562554 TI - Synchronous perforation of sigmoid diverticula: a rare presentation. AB - Diverticular disease affects more than 50% of the population over the age of 60 years in the west and becomes even more common as the population ages. Diverticulitis is one of the complications of diverticular disease and can culminate into colonic perforation. Though perforated diverticular disease is not uncommon, synchronous colonic perforations in diverticulitis is rare. Our patient was admitted with acute abdomen and exploratory laparotomy revealed two side-by side perforations of the sigmoid colon. A Hartmann's procedure was performed. Macro- and microscopic evaluation confirmed the presence of two perforated sigmoid diverticula due to diverticulitis. Simultaneous perforation of two abreast sigmoid diverticula is uncommon; thus, a cautious surgeon should always take into account such a probable diagnosis. PMID- 19562555 TI - Shrapnel injury due to a firecracker causing gastric and gallbladder perforation. AB - A 14-year-old boy presented with a penetrating injury to the abdomen after trying to light a firecracker. A piece of metal from the tin box in which he had ignited the firecracker had penetrated his abdomen. The patient, who was in a state of shock, underwent ultrasonography (USG) and radiography of the abdomen. USG revealed free fluid in the abdomen, while abdominal radiographs demonstrated free gas and a radiopaque object. Exploration revealed gastric and gallbladder perforations for which repair and cholecystectomy were done, respectively. Visceral injury following a firecracker explosion has not been reported previously. We would like to stress the importance of parental supervision to prevent irresponsible use of firecrackers, which can cause potentially life threatening visceral injuries. PMID- 19562556 TI - [Unusual migration of K-wire following fixation of clavicle fracture: a case report]. AB - The migration of various internal fixation devices, especially Kirshner (K) wires, is well established. The wires usually follow a retrograde path, protruding near the entry point. When they migrate in the other direction, serious problems may occur. Migration of K-wires to the lung, heart, spleen, subclavian artery, pulmonary artery and aorta have been reported in a few cases. A 26-year-old male with chest pain was seen in our clinic. The patient had been operated for left distal clavicle fracture two years before. No abnormality was noted on the physical examination. Radiographs showed migration of the wire outside the clavicle across the sternum to the opposite hemithorax. The pin was removed through the incision over the sternum under direct vision with thoracoscope. The pin was extrapleurally located. There was no additional morbidity attributed to thoracoscopy or chest tube. In conclusion, K-wires can easily migrate, resulting in serious complications. Close follow-up should be done after internal fixation. PMID- 19562557 TI - Management algorithm for intestinal obstruction due to ascariasis: a case report and review of the literature. AB - Ascaris lumbricoides is the largest and most prevalent of the human helminths. Ascariasis causes a unique type of intestinal obstruction with specific problems, having high morbidity and mortality. There is no management algorithm available in the literature to treat such cases. We aimed in this study to develop an algorithm for the management of intestinal obstruction due to ascariasis. A medline search of the English literature on the management of intestinal obstruction due to ascariasis was performed. Management of one of our patients with intestinal obstruction due to ascariasis is presented herein as an example of such cases. Intestinal obstruction due to ascariasis is a serious disease with a high morbidity and mortality, especially postoperatively. A management algorithm for the treatment was developed. The management of intestinal obstruction due to ascariasis was challenging and required difficult decisions. We think that this algorithm will be of great assistance in facilitating a rapid and successful treatment. This algorithm may prove to be of a great value for physicians treating such cases. PMID- 19562558 TI - Subtotal ear amputation with a very narrow pedicle: a case report and review of the literature. AB - Total ear amputation is common, and management can necessitate different procedures, especially microsurgical anastomosis. Partial ear amputations supplied by narrow pedicles, however, have been reported rarely. In a subtotally amputated auricle, the chance of survival depends on the vascularization within the pedicle. In our case, the right ear of a 36-year-old male patient was subtotally amputated following a traffic accident, leaving only a 6-mm skin pedicle on the cranial side. The subtotally amputated segment was bleeding from the wound margins. The ear was reattached with primary suture without using microsurgical techniques after optimal debridement. Postoperatively, we administrated dextran 40 for 5 days to improve the microcirculation and increase blood volume and antibiotic to control the infection. No signs of edema, venous congestion or arterial insufficiency were observed immediately after the operation or subsequently. The replanted auricle healed completely with 100% survival, resulting in an essentially normal contour and appearance. This successful result without microvascular anastomoses also points out the anatomical features of the auricular vascular networks. PMID- 19562559 TI - Is there any relation between development of persistent non-reassuring fetal heart rate pattern and acutely increased uterine artery vascular flow resistance during dinoprostone use in prolonged pregnancies? AB - OBJECTIVE: To investigate the relation between changes in Doppler parameters of fetal and uterine arteries and development of persistent non-reassuring fetal heart rate (FHR) pattern during induction of labor with dinoprostone (Propess) in pregnancies at >or=41 weeks gestation. DESIGN: Prospective cohort study. SETTING: Etlik Zubeyde Hanim Women's Hospital, Turkey. SAMPLE: One hundred forty-one prolonged pregnancies. METHODS: Doppler parameters of umbilical, middle cerebral, and uterine arteries were measured before and 4-6 hours after dinoprostone application between uterine contractions. Non-reassuring FHR pattern and persistent non-reassuring FHR pattern criteria were defined based on NICE 2007 guidelines. Women with successful spontaneous vaginal delivery were recruited as a control group (n=108), while women who underwent cesarean delivery due to persistent non-reassuring FHR pattern were recruited as a study group (n=15). MAIN OUTCOME MEASURES: Prediction of non-reassuring FHR pattern with Doppler analysis of uterine and fetal arteries. RESULTS: After dinoprostone application there was significant enhancement in uterine artery resistance index (RI) in the study group compared to the control group (p=0.002). Receiver operating characteristics curve analysis identified a uterine artery RI increase value of 0.11 as the optimal threshold for prediction of persistent non-reassuring FHR pattern with 73.3% sensitivity and 69.4% specificity. Logistic regression analysis demonstrated that an increase in the uterine artery RI was predictive for persistent non-reassuring FHR pattern (odds ratio (OR) 4.97; 95% CI 1.5 16.8). CONCLUSION: Acute increase in uterine artery RI due to dinoprostone use may end with persistent non-reassuring FHR pattern in prolonged pregnancies. This may allow earlier prediction of persistent non-reassuring FHR pattern development and risk assessment. PMID- 19562560 TI - Cryopreserved blastocysts have a lower implantation but an equal live birth rate as compared to fresh blastocysts of the same quality - a case-control study. AB - The purpose of this case-control study was to compare implantation, pregnancy, and live birth rate of cryopreserved embryos with that of fresh embryos. A total of 1,488 assisted reproductive technology (ART) trials including 188 cryopreserved embryo replacement trials were reviewed. After proofing the exclusion criteria, 112 patients undergoing a cryopreserved embryo replacement (study group) were matched with 112 patients undergoing a fresh embryo transfer (control group). Matching was done for age of patients, number, and quality of embryos (Day 5 embryos). Data concerning maternal characteristics, survival rate of warming procedure, implantation, pregnancy, ongoing pregnancy, and live birth rates were collected. A lower implantation rate was found for cryopreserved embryo replacement compared to transferring fresh embryos. Live birth rate was found to be equal in both groups. PMID- 19562561 TI - Association between aerobic fitness in late pregnancy and duration of labor in nulliparous women. AB - It is established that the level of physical activity in the population generally is too low, and data indicate that pregnant women are no exception. Studies of the effects of aerobic fitness on delivery outcomes are limited. In this observational study, we investigated the effect of aerobic fitness on duration of labor in nulliparous women who started labor spontaneously. Maximal oxygen uptake was measured in 40 nulliparous women at 35-37 weeks of gestation. Duration of labor was defined as the time between 3 cm cervical dilation with regular uterine contractions and delivery. The mean (SD) maximal oxygen uptake was 2.1 (0.3) L/min and duration of labor 583 (317) minutes. Duration of labor was inversely associated with maximal oxygen uptake after adjusting for birthweight (p = 0.034). We conclude that measurement of maximal oxygen uptake is safe in pregnancy at 35-37 weeks and that increased aerobic fitness was associated with shorter labor in nulliparous women who started labor spontaneously. PMID- 19562562 TI - Does repeat tibial tubercle osteotomy or intramedullary extension affect the union rate in revision total knee arthroplasty? A retrospective study of 74 patients. AB - BACKGROUND AND PURPOSE: Tibial tubercle osteotomy (TTO) is an established surgical technique for exposing the stiff kneein revision total knee arthroplasty (RTKA). The osteotomy is usually performed through the anterior metaphyseal cancellous bone of the tibia but it can be extended into the intramedullary canal if tibial stem and cement removal are necessary. Furthermore, repeat osteotomy may be required in another RTKA. We assessed whether intramedullary extension of TTO or repeat osteotomy affected the healing rate in RTKA. METHODS: We retrospectively evaluated 74 consecutive patients (39 women) with an average age of 60 (29-89) years who underwent 87 TTOs during RTKA. 1 patient had bilateral TTO.10 patients had repeat TTO and 1 patient received 3 TTOs in the same knee. The osteotomy was extramedullary in 57 knees and intramedullary in 30 knees. Osteotomy repair was performed with bicortical screws and/or wires. RESULTS: Bone healing occurred in all the cases. The median time to union was 15 (6-47) weeks. The median healing time for the extramedullary osteotomy group was 12 weeks and for the intramedullary osteotomy group it was 21 weeks (p = 0.002). Repeat osteotomy was not associated with delayed union. Neither intramedullary nor repeat osteotomy was found to increase the complication rate of the procedure. INTERPRETATION: Reliable bone healing can be expected with intramedullary extension or repeat TTO in RTKA. However, intramedullary extension of the osteotomy prolongs the union time of the tibial tubercle. PMID- 19562563 TI - Total elbow arthroplasty in rheumatoid arthritis: a population-based study from the Finnish Arthroplasty Register. AB - BACKGROUND AND PURPOSE: Although total elbow arthroplasty (TEA) is a recognized procedure for the treatment of the painful arthritic elbow, the choice of implant is still obscure. We evaluated the survival of different TEA designs and factors associated with survival using data from a nationwide arthroplasty register. METHODS: 1,457 primary TEAs for rheumatoid elbow destruction were performed during 1982 to 2006 in one hospital specialized in the treatment of rheumatoid arthritis (n = 776) and in 19 other hospitals (n = 681). The mean age of the patients was 59 years and 87% of the TEAs were performed in women. We selected different contemporary TEA designs, each used in more than 40 operations including the Souter-Strathclyde (n = 912), i.B.P./Kudo (n = 218), Coonrad-Morrey (n = 164), and NESimplavit/Norway (n = 63) to assess their individual survival rates. Kaplan-Meier analysis and the Cox regression model were used for survival analysis. RESULTS: The most frequent reason for revision was aseptic loosening (47%). We found no differences in survival rates between different TEA designs. We did, however, find a 1.5-fold (95% CI: 1.1-2.1) elevated risk of revision in unspecialized hospitals as compared to the one hospital specialized in treatment of rheumatoid arthritis. In the Souter-Strathclyde subgroup, there was a reduced risk of revision (RR 0.6, p = 0.001) in TEAs implanted over 1994-2006 as compared to those implanted earlier (1982-1993). The 10-year survivorship for the whole TEA cohort was 83% (95% CI: 81-86), which agrees with earlier reports. INTERPRETATION: The influence of implant choice on the survival of TEA is minor compared to hip and knee arthroplasties. Inferior survival rates of the TEAs performed in the unspecialized hospitals demonstrates the importance of proper indications, surgical technique, and postoperative follow-up, and endorses the need for centralization of these operations at specialized units. PMID- 19562564 TI - Additive fiber-cerclages in proximal humeral fractures stabilized by locking plates: no effect on fracture stabilization and rotator cuff function in human shoulder specimens. AB - BACKGROUND AND PURPOSE: The effect of additive fiber-cerclages in proximal humeral fractures stabilized by locking plates on fracture stabilization and rotator cuff function is unclear. Here it was assessed in a human cadaver study. METHODS: 24 paired human shoulder specimens were harvested from median 77-year old (range 66-85) female donors. An unstable 3-part fracture model with an intact rotator cuff was developed. 1 specimen of each pair received an additive fiber cerclage of the rotator cuff after plate fixation, and the other one received a plate fixation without an additive fiber-cerclage. Force-controlled hydraulic cylinders were used to simulate physiological rotator cuff tension, while a robot assisted shoulder simulator performed 4 relevant cases of load: (1) axial loading at 0 degrees, (2) glenohumeral abduction at 60 degrees, (3) internal rotation at 0 degrees abduction, and (4) external rotation at 0 degrees abduction, and imitated hanging arm weight during loading without affecting joint kinematics. A 3-dimensional real-time interfragmentary motion analysis was done in fracture gaps between the greater tuberosity and the head, as well as subcapital. The capacity of the rotator cuff to strain was analyzed with an optical system. RESULTS: Interfragmentary motion was similar between the groups with and without fiber-cerclages, in both fracture gaps and in any of the cases of load. Cerclages did not impair the capacity of the rotator cuff to strain. INTERPRETATION; Provided that unstable 3-part fractures are reduced and stabilized anatomically by a locking plate, additive fiber-cerclages do not reduce interfragmentary motion. Additive fiber-cerclages may be necessary in locking plate osteosyntheses of multiple-fractured greater tuberosities or lesser tuberosity fractures that cannot be fixed sufficiently by the plate. PMID- 19562565 TI - Periprosthetic DXA after total hip arthroplasty with short vs. ultra-short custom made femoral stems: 37 patients followed for 3 years. AB - BACKGROUND AND PURPOSE: Dual-energy X-ray absorptiometry (DXA) analysis of the 7 periprosthetic Gruen zones is the most commonly used protocol to evaluate bone remodeling after the implantation of conventional femoral stems. We assessed the value of DXA after cementless primary total hip arthroplasty (THA) by comparing the effect of progressive shortening of the stem of two femoral implants on periprosthetic bone remodeling using a specifically developed protocol of analysis with 5 periprosthetic regions of interest (ROIs). PATIENTS AND METHODS: Bone mineral density (BMD) was evaluated in 37 patients in the plateau stage, 3 years after THA. Two femoral implants featuring conceptually new designs and surgical technique were tested: types 1 and 2, characterized by extremely short stem and virtual absence of distal stem, respectively. RESULTS: We found that progressive shortening of the femoral stem produces more proximal loading, which effectively preserves metaphyseal bone stock and increases periprosthetic BMD in the medial ROIs over time. In the type 2 group, higher absolute BMD values were observed in medial ROIs 4 and 5. No differences were found in ROIs 1, 2, and 3. INTERPRETATION: This study shows the flexibility of DXA in adapting the protocol of periprosthetic analysis to the specific requirements of new implant designs, and it shows its high sensitivity in evaluation of the biological response of bone to changes in implant shape. PMID- 19562566 TI - Obstructive left-sided prosthetic valve thrombosis. AB - Obstructive prosthetic valve thrombosis is a serious complication in patients with prosthetic heart valves. It should be suspected in patients with worsening functional class, embolic phenomena, and inadequate anticoagulation. TTE is very informative and the most frequent modality to begin with. TEE is extremely important, with its unique role in excluding high-risk thrombi. Fluoroscopy is currently the best modality for the assessment of leaflet motion, especially for the aortic position, while cardiac CT may be of added value due to its unique post-processing features. Efforts to discriminate thrombosis from pannus should be made, although current methods are frequently inconclusive. The application of 3-dimensional TEE is hoped to improve the diagnostic accuracy further. Once the diagnosis is established, therapy should be offered according to the local expertise, considering the risk of surgery, the risk of thrombolysis (mainly- bleeding and embolism), the patients' functional class, and the likelihood of achieving valve reopening. Guidelines are numerous and puzzling. Thrombus size is probably the most important determinant of complications, and if it is small, thrombolysis is probably advised across all degrees of functional class, as suggested by American College of Chest Physicians. PMID- 19562567 TI - Growth and chronic disease: findings in the Helsinki Birth Cohort. AB - There is now clear evidence that the pace and pathway of early growth is a major risk factor for the development of a group of chronic diseases that include coronary heart disease, stroke, type 2 diabetes and hypertension. This has led to a new 'developmental' model for these disorders. The so-called 'fetal origins hypothesis' proposes that the disorders originate through developmental plasticity, whereby malnutrition during fetal life, infancy and early childhood permanently change the structure and function of the body, a phenomenon known as 'programming'. This paper reviews recent findings in the Helsinki Birth Cohort, which comprises 13 345 men and women born in the city during 1934-1944. There is also an older cohort comprising 7086 people born during 1924-1933. We review the paths of pre- and postnatal growth that lead to later disease. Children who later develop coronary heart disease and type 2 diabetes grow slowly during fetal life and infancy but thereafter increase their body mass indices rapidly. Those who later develop stroke grow slowly in fetal life, infancy and during childhood. We also review how the growth of girls during infancy, childhood and at puberty influences chronic disease in the next generation. PMID- 19562568 TI - Comparisons of collagen gel and collagen matrix as cell growth microenvironment: the physical-chemical properties. AB - To evaluate the changes of gel scaffold and spongy scaffold in fibroblast culture in vitro, two kinds of collagen dermis were constructed in this research. Type I collagenase and Dispase were used to isolate dermis fibroblast from neonate prepuce. The gel dermis was constructed by mixing the fibroblast and collagen swelling solution, and the collagen spongy dermis was also constructed. After histological and immunohistochemical staining, these two dermis' properties were compared. Compared with gel dermis, the spongy dermis possessed more superior tension strength, but it decreased significantly during the culture. The transition temperature in DSC reduced also. The rupture length of gel dermis increased, on the contrary. Gel dermis contained more moisture than spongy dermis. It had been observed that the collagen gel was more suitable for fibroblast in maintaining its morphology than spongy scaffold, though with less cell number. The reason might be due to the special structure of collagen gel. Water in gel is sub-bound water, which is different from free water, and it is suitable for fibroblasts to secrete extracellular matrix. It could be concluded that the collagen gel dermis is better than collagen spongy dermis in the biological property. PMID- 19562570 TI - Hematoxylin shortages: their causes and duration, and other dyes that can replace hemalum in routine hematoxylin and eosin staining. AB - The origins of repeated hematoxylin shortages are outlined. Lack of integration in the hematoxylin trade exacerbates the problems inherent in using a natural product. Separate corporations are engaged in tree growth and harvesting, dye extraction, processing of extracts to yield hematoxylin, and formulation and sale of hematoxylin staining solutions to the end users in biomedical laboratories. Hematoxylin has many uses in biological staining and no single dye can replace it for all applications. Probably, the most satisfactory substitutes for aluminum hematoxylin (hemalum) are the ferric complexes of celestine blue (CI 51050; mordant blue 14) and eriochrome cyanine R (CI 43820; mordant blue 3, also known as chromoxane cyanine R and solochrome cyanine R). The iron-celestine blue complex is a cationic dye that binds to nucleic acids and other polyanions, such as those of cartilage matrix and mast cell granules. Complexes of iron with eriochrome cyanine R are anionic and give selective nuclear staining similar to that obtained with acidic hemalum solutions. Iron complexes of gallein (CI 45445; mordant violet 25), a hydroxyxanthene dye, can replace iron-hematoxylin in formulations for staining nuclei, myelin, and protozoa. PMID- 19562571 TI - Expression of Notch family members in cultured murine articular chondrocytes. AB - The Notch family is involved in cell differentiation during embryogenesis. Osteoarthritic chondrocytes undergo morphological and biochemical changes leading to the de-differentiation process. In the study reported here, we were interested in the involvement of the Notch pathway in murine articular chondrocyte de differentiation. Articular chondrocytes were subjected to several cell culture passages and treated with or without a Notch inhibitor, N-[N-(3, 5 difluorophenacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester (DAPT). Chondrocyte morphology was studied using optical microscopy. Immunocytochemistry and immunoblot were performed to study the expression of collagens and Notch family members. Without DAPT treatment, chondrocyte de-differentiation resulted in fibroblast-like morphology. This was confirmed by immunocytochemical staining and immunoblot analysis, which showed an increase in collagen type I (col I) and a decrease in collagen type II (col II) expression. With DAPT treatment, de differentiation was delayed. Immunocytochemistry and immunoblot analysis showed during the first passages inhibition of col II expression, which then was re instituted during the last passage, suggesting chondrocyte re-differentiation. In the study reported here, we showed that inhibition of the Notch receptor not only delayed the de-differentiation process, but also chondrocyte re-differentiation, which confirms the involvement of the Notch pathway in chondrocyte de differentiation. PMID- 19562573 TI - Pulse pressure, left ventricular function and cardiovascular events during antihypertensive treatment (the LIFE study). AB - BACKGROUND: Pulse pressure (PP) has been related to risk of cardiovascular events in hypertension. However, less is known about modification of this risk marker during antihypertensive treatment in patients with left ventricular (LV) hypertrophy. METHODS: Associations of in-treatment PP with LV systolic function and cardiovascular events was assessed in 883 patients with electrocardiographic LV hypertrophy during 4.8 years of randomized losartan- or atenolol-based treatment within the echocardiographic substudy of the Losartan Intervention For Endpoint reduction in hypertension (LIFE) study. RESULTS: PP was similarly reduced by both treatments. In different multiple regression models, lower in treatment PP was independently associated with lower in-treatment LV ejection fraction (beta=0.16), stress-corrected midwall shortening (beta=0.20), stroke volume (beta=0.11) and cardiac index (beta=0.07, all p<0.05). In time-varying Cox regression models, 10 mmHg lower in-treatment PP was associated with a 28% higher rate of cardiovascular events [hazard ratio, HR = 1.28 (1.09 - 1.52), p<0.01] independent of in-treatment LV mass and ejection fraction, history of ischemic heart disease, Framingham risk score and study treatment allocation. CONCLUSION: During systematic antihypertensive treatment in hypertensive patients with electrocardiographic LV hypertrophy, lower in-treatment PP was associated with lower in-treatment LV function and cardiac output as well as higher rate of cardiovascular events. PMID- 19562574 TI - Effects of low-dose liquorice alone or in combination with hydrochlorothiazide on the plasma potassium in healthy volunteers. AB - Both liquorice and thiazide diuretics have hypokalaemic effects. In spite of their prevalent use, there are no studies on hypokalaemia caused by the combination of liquorice and thiazides. We recruited 10 healthy volunteers in order to study the effects of 32 g liquorice alone or in combination with 25mg hydrochlorothiazide a day for 2 weeks. The trial had a randomized, open and crossover design. During the liquorice phase, there were no changes in plasma potassium, sodium, creatinine, renin activity, serum aldosterone, blood pressure or heart rate. Weight tended to increase by 0.4 kg (70.2 to 70.6 kg; p=0.056). During the liquorice-hydrochlorothiazide phase, the plasma potassium decreased by 0.32 mmol/l (p=0.0015), plasma renin activity increased by 1.6 microg/l/h (p=0.0064) and the weight decreased by 0.9 kg (70.5 to 69.6 kg, p=0.0065). Twenty per cent of the subjects (2/10) became hypokalaemic during the combined liquorice hydrochlorothiazide treatment. Furthermore, both subjects developed hypokalaemia within the first week of the combined treatment leading to premature discontinuation. The evaluation of liquorice consumption habits is warranted when initiating thiazide medications to avoid the excessive risk of hypokalaemia associated with the combined use of low-dose liquorice and thiazide diuretics. PMID- 19562575 TI - Natriuretic peptides as indicators of cardiac remodeling in hypertensive patients. AB - AIMS: This study was performed to evaluate the relationship between three different natriuretic peptides and left ventricular mass, function and diameter, and kidney function in patients with hypertension. METHODS: One hundred and thirty-nine patients with moderate hypertension were consecutively included. N terminal brain natriuretic peptide (Nt-BNP), brain natriuretic peptide (BNP) and N-terminal pro-atrial natriuretic peptide (Nt-ANP) were analyzed. Cardiac remodeling was assessed by echocardiography (UCG) and glomerular filtration was estimated by cystatin C. RESULTS: Patients were stratified into four groups with regard to the extent of cardiac remodeling: (1) no remodeling; (2) one of left ventricular hypertrophy, left ventricular dysfunction or left ventricular dilatation; (3) two of above and (4) all three parameters. All peptides differed significantly between the groups (all p<0.001), with a continuous stepwise increase from groups 1 through 4. Receiver operating characteristic analysis showed equal diagnostic performances for the detection of any cardiac abnormalities for Nt-BNP [area under curve, AUC=0.63 (0.52-0.75), p=0.026] and BNP [AUC=0.64 (0.53-0.76), p=0.019], both, however superior to Nt-ANP [AUC=0.59 (0.47-0.70), p=0.139]. In multivariable linear regression analysis, all three indicators of cardiac remodeling were independently correlated with ln Nt-BNP and ln BNP, whereas only left ventricular diameter was independently correlated with ln Nt-ANP. CONCLUSIONS: Natriuretic peptide levels increased with increasing number of markers of cardiac remodeling. Nt-BNP and BNP are useful to discriminate between patients with regard to cardiac remodeling and might be considered a screening tool in order select patients eligible for further examination with UCG examination. PMID- 19562576 TI - Mesenchymal stem cells as a gene therapy carrier for treatment of fibrosarcoma. AB - BACKGROUND AIMS: Cell-based gene therapy is an alternative to viral and non-viral gene therapy. Emerging evidence suggests that mesenchymal stem cells (MSC) are able to migrate to sites of tissue injury and have immunosuppressive properties that may be useful in targeted gene therapy for sustained specific tissue engraftment. METHODS: In this study, we injected intravenously (i.v.) 1x10(6) MSC, isolated from green fluorescent protein (GFP) transgenic rats, into Rif-1 fibrosarcoma-bearing C3H/HeN mice. The MSC had been infected using a lentiviral vector to express stably the luciferase reporter gene (MSC-GFP-luci). An in vivo imaging system (IVIS 200) and Western blotting techniques were used to detect the distribution of MSC-GFP-luci in tumor-bearing animals. RESULTS: We observed that xenogenic MSC selectively migrated to the tumor site, proliferated and expressed the exogenous gene in subcutaneous fibrosarcoma transplants. No MSC distribution was detected in other organs, such as the liver, spleen, colon and kidney. We further showed that the FGF2/FGFR pathways may play a role in the directional movement of MSC to the Rif-1 fibrosarcoma. We performed in vitro co-culture and in vivo tumor growth analysis, showing that MSC did not affect the proliferation of Rif-1 cells and fibrosarcoma growth compared with an untreated control group. Finally, we demonstrated that the xenogenic MSC stably expressing inducible nitric oxide synthase (iNOS) protein transferred by a lentivirus-based system had a significant inhibitory effect on the growth of Rif-1 tumors compared with MSC alone and the non-treatment control group. CONCLUSIONS: iNOS delivered by genetically modified iNOS-MSC showed a significant anti-tumor effect both in vitro and in vivo. MSC may be used as a target gene delivery vehicle for the treatment of fibrosarcoma and other tumors. PMID- 19562579 TI - Patient centered goal-setting in a subacute rehabilitation setting. AB - PURPOSE: The purpose of this research is to describe current practices in goal setting within a subacute rehabilitation setting from the perspective of therapists representing the disciplines of occupational therapy, speech pathology and physiotherapy. METHOD: Qualitative semi-structured email interviews were conducted with therapists from the Geriatric Assessment and Rehabilitation Unit of an Australian hospital. Therapists were required to respond to questioning with reference to identified rehabilitation patients with stroke. RESULTS: Three approaches to goal-setting were identified: therapist controlled, therapist led and patient centred. Goals aimed at the ICF levels of impairment and activity limitations were predominant. Barriers to a patient centered goal-setting approach largely outweighed facilitators. Potential successful resolutions were offered to overcome these barriers. CONCLUSIONS: The inability of patients to participate fully in the goal-setting process largely determines the approach taken by therapists. This influences the level of patient centeredness incorporated into the goal-setting process. Goals expressed at the level of impairment, by therapists, may be stepping stones to perceived patient goals at the levels of activity and participation. Barriers to a patient centered approach can be overcome through education of the patient and family regarding the nature of the injury and modification of communication between therapist and patient. PMID- 19562584 TI - The break remains - elderly people's experiences of a hip fracture 1 year after discharge. AB - PURPOSE: To explore experienced long-term consequences of a hip fracture and conceptions of what influences hip fracture recovery among community-living elderly people 1 year after discharge. METHOD: Fifteen subjects (13 females and 2 males), aged 66-94, were interviewed. The phenomenographic method was used for analysis. RESULTS: Experiences of insecurity and restricted life dominated the interviews. The descriptive categories within experienced consequences of a hip fracture were: (1) isolated life with more restricted activity and fewer social contacts, with the two sub-categories (a) more insecure and afraid and (b) more limited ability to move, (2) disappointed and sad that identity and life have changed and (3) satisfied with the situation or feeling even better than before the fracture. The categories within conceptions of what influences hip fracture recovery were: (4) own mind and actions influence recovery, (5) treatment and actions from others influence recovery and (6) you cannot influence recovery. CONCLUSION: The findings accentuate that the negative consequences of a hip fracture are substantial and long-lasting. As it strikes mostly elderly people, who may have experienced earlier losses and growing disabilities, a hip fracture could add to the risk of losing important life values. Furthermore, the findings indicate that all health care professionals who meet the patients need to consider the patients' own experiences and possible fear and not merely focus on the physical injury and disabilities. PMID- 19562586 TI - Medicalising disability? Regulation and practice around fitness assessment of disabled students and professionals in nursing, social work and teaching professions in Great Britain. AB - PURPOSE: The reliance on medical information and on occupational health (OH) professionals in ascertaining fitness of applicants and registrants within the educational and employment contexts may lead to the medicalisation of disability. The Disability Rights Commission's Formal Investigation into the regulation of three public sector professions of nursing, social work and teaching in Britain sheds light on the nature of regulatory fitness requirements and how these are implemented in practice. METHOD: The multi-pronged investigation included a review of relevant statutory and regulatory frameworks, formal written and oral evidence submitted by key stakeholder organisations and research into formal and informal fitness assessments within the education and employment contexts. RESULTS: There are varied and vague fitness requirements in all three professions. OH professionals figure prominently in formal and informal decision making around fitness within education and employment settings, regardless of regulatory prescriptions. There is a multitude of approaches. There are, however, particular issues in the deployment of OH expertise within the employment setting. CONCLUSIONS: The determination of fitness should not rely solely on medical information. Blanket fitness requirements that are not contextualized against specific competencies for particular jobs are inappropriate. More collaborative and integrated working is necessary, particularly in exploring how reasonable adjustments may be provided to enable safe and effective practice. The positive spirit of the disability equality duty should be embraced. PMID- 19562587 TI - Women with disabilities' experiences of government employment assistance in Canada. AB - PURPOSE: The purpose of this article is to explore women with disabilities' experiences of government employment assistance in Canada. METHOD: The article draws on the results of an online survey conducted in 2006. Data were coded and analysed according to key themes. RESULTS: The results indicate that many of the women with disabilities who responded to the survey regarded the employment assistance they have received as of very limited importance to their abilities to find and keep paid work. Their comments suggest that existing forms of government employment assistance, such as the provision of wage subsidies to employers, may be of limited effectiveness in enabling at least some women with disabilities to find and keep paid work. The women's comments point to the limitations of existing employment assistance strategies including the need to shift program emphases away from individualistic models aimed at 'reforming the worker' and toward systemic barriers to employment. CONCLUSIONS: The article concludes by discussing the implications of survey results for future research and strategies for improving the types of employment assistance available to women with disabilities. PMID- 19562588 TI - Workshop-based methodology to understand the risks in grain export inspection and certification. AB - Much of the human factors contribution in risk assessment and risk management has been focused on systems or product safety; the profession has a much smaller research base regarding risks to do with regulation, certification and public policy, for example. This paper discusses an explicitly human factors contribution to understanding and managing risk for the inspection and export certification of grain and plant products in Australia. Training and awareness workshops, incorporating elements of focus groups, were run for 12 groups of staff and managers from the government department concerned. As well as training in risk management the workshops were used to come to an understanding of the work of the inspectors and other staff, to identify the sources of risk to the successful completion of their work and to develop the basis for a risk assessment framework and tool. The paper is methodological in focus and describes the development and running of the workshops and explains how a human factors oriented risk register was developed on the basis of identification of potential threats and errors in the system. Whilst the contribution of ergonomics is increasingly important as regards safety risk assessment, professionals have been less active as regards business, public policy and even engineering risk. This paper describes an approach within which a new domain was studied and the risks of all kinds identified, preparatory to development of a risk assessment tool. PMID- 19562589 TI - Understanding safety and production risks in rail engineering planning and protection. AB - Much of the published human factors work on risk is to do with safety and within this is concerned with prediction and analysis of human error and with human reliability assessment. Less has been published on human factors contributions to understanding and managing project, business, engineering and other forms of risk and still less jointly assessing risk to do with broad issues of 'safety' and broad issues of 'production' or 'performance'. This paper contains a general commentary on human factors and assessment of risk of various kinds, in the context of the aims of ergonomics and concerns about being too risk averse. The paper then describes a specific project, in rail engineering, where the notion of a human factors case has been employed to analyse engineering functions and related human factors issues. A human factors issues register for potential system disturbances has been developed, prior to a human factors risk assessment, which jointly covers safety and production (engineering delivery) concerns. The paper concludes with a commentary on the potential relevance of a resilience engineering perspective to understanding rail engineering systems risk. Design, planning and management of complex systems will increasingly have to address the issue of making trade-offs between safety and production, and ergonomics should be central to this. The paper addresses the relevant issues and does so in an under-published domain - rail systems engineering work. PMID- 19562590 TI - Predicting vigilance: a fresh look at an old problem. AB - Individual differences in vigilance are ubiquitous and relevant to a variety of work environments in industrial, transportation, medical and security settings. Despite much previous work, mostly on personality traits, it remains difficult to identify vigilant operators. This paper reviews recent research that may point towards practically useful predictor variables for vigilance. Theoretical approaches to identifying predictors that accommodate the heterogeneous nature of vigilance tasks are compared. The article surveys recent empirical studies using personality measures, ability tests and scales for stress and coping as predictors of vigilance. Promising new constructs include trait scales linked to fatigue, abnormal personality and the stress state of task engagement. Implications of the data reviewed for occupational selection are discussed. Selection should be based on a multivariate assessment strategy, cognitive task analysis of the operational vigilance task and use of work sample measures to capture typical stress responses to the task. This review paper surveys recent research that may point towards practically useful predictor variables for vigilance. The article surveys recent empirical studies using personality measures, ability tests and scales for stress and coping as predictors of vigilance. Selection should be based on a multivariate assessment strategy. PMID- 19562591 TI - Prospective risk analysis of health care processes: a systematic evaluation of the use of HFMEA in Dutch health care. AB - The aim of this study was to evaluate the use of Healthcare Failure Mode and Effect Analysis (HFMEA) in Dutch health care by means of user feedback. Thirteen HFMEA analyses of various health care processes were successfully concluded and on average took 69 person-hours (excluding reporting). These results show that HFMEA can successfully be applied in Dutch health care. However, the user feedback also uncovered several perceived drawbacks, such as the fact that HFMEA is very time-consuming and that, particularly, the risk assessment part of HFMEA is difficult to carry out. Moreover, a lack of guidance with regard to the identification of failure mode causes and effective actions might influence the quality of the outcomes of an HFMEA analysis. Several suggestions are put forward to improve the perceived utility and acceptance of HFMEA. Nevertheless, future research is necessary to evaluate the actual effects of these recommendations. Error modelling and risk analysis, and their contribution to explaining human performance in socio-technical systems, traditionally belong to the field of ergonomics. The user feedback on HFMEA and the suggestions that are put forward may also be useful for (H)FMEA and hazard analysis and critical control point applications in sectors other than health care. PMID- 19562592 TI - Inter-rater reliability of PATH observations for assessment of ergonomic risk factors in hospital work. AB - This study examined the inter-rater reliability of expert observations of ergonomic risk factors by four analysts. Ten jobs were observed at a hospital using a newly expanded version of the PATH method (Buchholz et al. 1996), to which selected upper extremity exposures had been added. Two of the four raters simultaneously observed each worker onsite for a total of 443 observation pairs containing 18 categorical exposure items each. For most exposure items, kappa coefficients were 0.4 or higher. For some items, agreement was higher both for the jobs with less rapid hand activity and for the analysts with a higher level of ergonomic job analysis experience. These upper extremity exposures could be characterised reliably with real-time observation, given adequate experience and training of the observers. The revised version of PATH is applicable to the analysis of jobs where upper extremity musculoskeletal strain is of concern. PMID- 19562593 TI - Car driving with and without a movable back support: Effect on transmission of vibration through the trunk and on its consequences for muscle activation and spinal shrinkage. AB - The aim of this study was to test the effect of a movable backrest on vibration transmission through the trunk during driving and on the physiological consequences thereof. Eleven healthy male subjects drove for about 1 h on normal roads with a movable and with a fixed backrest while surface electromyography (EMG) was measured at the level of the fifth lumbar vertebra (L5) and vertical accelerations were measured at the seat, backrest and at the spine at the levels of the second sacral vertebra (S2) and seventh cervical vertebra (C7). The movable backrest significantly reduced accelerations at C7 by up to 11.9% at the 5 Hz frequency band. The movable backrest also significantly reduced the coherence and transmission between S2 and C7 accelerations, but not the differential motion between these sensors. EMG at both sides of L5 was on average 28% lower when using the movable backrest. Spinal shrinkage was unaffected by backrest type. It is concluded that a movable backrest reduces the transmission of vibration through the trunk and that it reduces low back EMG. Car driving is associated with the risk of developing low back pain and this may be related to exposure to whole body vibration. This study found an effect of a simple ergonomics measure on the transmission of vibration through the trunk as well as on back muscle activation. PMID- 19562594 TI - Dependence of safety margins in grip force on isometric push force levels in lateral pinch. AB - This study examined the relationship between safety margin and force level during an isometric push task in a lateral pinch posture. Ten participants grasped an object with an aluminium- or rubber-finished grip surface using a lateral pinch posture and exerted 20%, 40%, 60%, 80% and 100% of maximum push force while voluntary grip force was recorded. Then minimum required grip force was measured for each push force level. Mean safety margin, the difference between voluntary and minimum required grip forces, was 25% maximum voluntary contraction (MVC) when averaged for all push levels. Safety margin significantly increased with increasing push force for both grip surfaces. Grip force used during maximum push exertion was only 74% lateral pinch grip MVC. Possible underlying mechanisms for increasing safety margin with increasing push force are discussed as well as the implication of this finding for ergonomic analysis. This study demonstrates that ergonomic analyses of push tasks that involve friction force should account for safety margin and reduced grip strength during the push. Failure to consider these can result in overestimation of people's push capability. PMID- 19562595 TI - Effects of strap support in a hand-held device on the muscular activity in female workers assessed by electromyography and subjective rating. AB - The present study evaluates the potential mitigation of physical workload when using strap support for a portable device. The experiments were designed as consecutive sessions over a 2-h period. Electromyogram signals were recorded from four muscles of six subjects. The perceived level of fatigue on the whole body as well as in the shoulder, arm, lower back and legs was assessed using Borg's CR-10 scale. All subjects were tested under eight experimental conditions. Results indicated that the biceps brachii muscle displayed significantly lower activity with strap support than without a strap. In the experiments with and without a strap, different levels of force were imposed on the various muscles, which caused changes in the distribution of the physical load. Although the role of the strap might seem evident, using strap support did not always decrease the sensation of fatigue. However, for short-term tasks, using a strap may be recommended. PMID- 19562596 TI - Reliability of different thresholds for defining muscular rest of the trapezius muscles in computer office workers. AB - This study aimed at documenting the reliability of different thresholds used for defining the muscular rest of the trapezius muscles of 27 computer office workers, using surface electromyography (EMG) signals collected in the field. Measurement strategies for increasing the reliability of the results were also explored. Ten different thresholds to define muscular rest were compared: 1) five normalised (individualised) thresholds; 2) three absolute thresholds (in muV); 3) two absolute but individualised thresholds. The reliability was assessed using both a 15-min standardised computer task and 45 min of regular computer work. The main findings were: 1) overall, in a repeated measures study design, muscular rest variables were more reliable with the use of absolute thresholds when compared to normalised and individualised thresholds; 2) excellent reliability (index of dependability >0.75) can be reached when averaging the scores over 2 days; 3) using a standardised task instead of regular work does not necessarily lead to more reliable results. PMID- 19562597 TI - The effect of idle time thresholds on computer use time estimations by electronic monitoring. AB - This study examined the effect of idle time setting on the estimation of computer use times by electronic activity monitoring and validated its use by comparing it with video record observations. Twenty-four study subjects were recruited and their work with computers was monitored for 1 h. With the estimates by video record observation as references, the best idle time settings for electronic activity monitoring with the least relative errors were 25, 2.5 and 2.5 s, respectively, for total computer, keyboard and mouse use time estimations. These estimates were highly correlated with the corresponding references (r = 0.918 0.964, p < 0.0001), accompanied by limited mean estimate differences ranging from -3.0 +/- 2.8% to 1.3 +/- 1.6%. The estimates by self-report were moderately correlated with the corresponding references (r = 0.387-0.678), with greater mean estimate differences. This study concluded that, for electronic activity monitoring methods, the most appropriate thresholds for idle time setting are 25 s, 2.5 s and 2.5 s for total computer, keyboard and mouse use time estimates, respectively. This method may help evaluate physical work-loading with computer works through a large-scale epidemiological study. PMID- 19562598 TI - The advantage of positive text-background polarity is due to high display luminance. AB - Reading text from computer screens is better when text is printed in dark letters on light background (positive polarity) than when it is printed in light letters on dark background (negative polarity). An experiment is presented that tests whether this positive polarity advantage is due to the fact that overall display luminance is typically higher for positive than for negative polarity displays. To this end, text-background polarity and display luminance were manipulated independently. No positive polarity advantage was observed when overall display luminance of positive and negative polarity displays was equivalent. There was only an effect of display luminance, with better performance for the higher luminance displays. This suggests that the positive polarity advantage is in fact due to the typically higher luminance of positive polarity displays. Readability of text presented on computer screens (e.g. on websites) is better when the overall display luminance level is high, as in positive polarity displays (dark letters on light background). Display polarity per se does not affect readability. PMID- 19562601 TI - Direct, real-time monitoring of superoxide generation in isolated mitochondria. AB - Mitochondria are one of the major sources of reactive oxygen species (ROS) in mammalian cells. The generation of ROS underlies many physiological and pathophysiological processes that occur within cellular systems. Superoxide ([image omitted] ) is the proximal ROS generated during electron 'leakage' from the mitochondrial electron transport chain (mETC) and is known to be released at mitochondrial complex I and complex III. Monitoring mitochondrial [image omitted] production directly and in real-time offers the potential to improve understanding of the complex mechanisms involved during mitochondrial [image omitted] generation. This study reports the novel application of a cytochrome c functionalized amperometric sensor for monitoring [image omitted] generation in isolated mitochondrial fractions. The non-invasive sensor system described allowed a comparison of [image omitted] production following specific inhibition of complex I and complex III of the mETC to be made directly and in real-time. PMID- 19562603 TI - The levonorgestrel intrauterine system as an alternative to hysterectomy for the treatment of idiopathic menorrhagia. AB - AIM OF THE STUDY: Prospective, observational study to determine the percentage of hysterectomies cancelled after a year of treatment with levonorgestrel intrauterine system (LNG-IUS) among women diagnosed with idiopathic menorrhagia. MAIN FINDINGS: Eighty-two women with a mean age of 44.3 + or - 4.9 were enrolled. Throughout 1-year follow-up, progressive and significant reduction was observed in number of days of bleeding (8.9 + or - 4.0 vs. 5.0 + or - 5.4), number of sanitary measures (29.3 + or - 19.4 vs. 8.1 + or - 10.8) and percentage of patients having intense/very intense bleeding (98.8%vs. 6.4%). Duration of menstrual cycle significantly increased from 26.9 + or - 5.5 to 52.6 + or - 33.6 days. Significant improvement in overall health-related quality of life was achieved. Patient satisfaction was good/very good in 70.7%. Considering only women who attended 12-month visit satisfaction reached 91.2%. 75.6% of scheduled hysterectomies were cancelled. Adverse effects were recorded in less than 40% of patients with no significant differences between visits. Adverse effects led to premature discontinuation of treatment in seven cases. No serious adverse effects were encountered. INTERPRETATION OF RESULTS: LNG-IUS meets the effectiveness and tolerability criteria for being considered as a first choice treatment option for women with idiopathic menorrhagia. Its use may contribute to decrease the large number of hysterectomies scheduled in Spain. PMID- 19562604 TI - The fertility-sparing treatment in patients with endometrial atypical hyperplasia and early endometrial cancer: a debated therapeutic option. AB - Fertility-sparing treatment may represent a realist option for accurately selected young patients with endometrial atypical hyperplasia or well differentiated, early endometrial cancer. Oral progestins, and especially medroxyprogesterone acetate (MPA) and megestrol acetate with different doses and schedules, represent the most commonly used hormone agents in this clinical setting. Approximately three fourths of the women achieve a histologically documented complete response, with an mean response time of 12 weeks, but about one third of these subsequently developed a recurrence after a mean time of 20 months. The expression of receptor for progesterone receptor (PR), PTEN gene, DNA mismatch repair gene MLH1 and phospho-AKT on tissue specimens may be useful for selecting patients fit for a conservative management. Several successful pregnancies have occurred after a fertility-sparing treatment of endometrial atypical hyperplasia or endometrial cancer, more frequently with assisted reproductive technologies. The implementation of in vitro fertilisation techniques not only increases the chance of conception, but it may also decrease the interval to conception. The opportunity of a demolitive surgery after delivery or after childbearing being no longer required is a still debated issue. Large multicenter trials are strongly warranted to better define the selection criteria for a conservative treatment, endocrine regimen of choice, the optimal dosing, the duration of treatment and follow-up protocols. In any case, the patient should be accurately informed about the relatively high recurrence rates after complete response to hormone treatment and expectations for pregnancy. PMID- 19562607 TI - Studies on effect of multiple heating/cooling cycles on the resistant starch formation in cereals, legumes and tubers. AB - 'Resistant starch' (RS) is defined as starch and starch degradation products that resist the action of amylolytic enzymes. The effect of multiple heating/cooling treatments on the RS content of legumes, cereals and tubers was studied. The mean RS contents of the freshly cooked legumes, cereals and tubers (4.18%, 1.86% and 1.51% dry matter basis, respectively) increased to 8.16%, 3.25% and 2.51%, respectively, after three heating/cooling cycles (P< or =0.05) with a maximum increase of 114.8% in pea and a minimum of 62.1% in sweet potato (P< or =0.05). Significant positive correlations were observed between the RS content and amylose (y=0.443x-5.993, r=0.829, P< or =0.05, n=9) as well as between the percentage increase in RS and insoluble dietary fiber content (y=2.149x-24.787, r=0.962, P< or =0.05, n=9). A differential scanning calorimeter study showed an increase in the T(0), T(p), T(c) and DeltaH values of the repeatedly autoclaved/cooled starches. The intact granular structure was also observed disappear, as studied using scanning electron microscopy. PMID- 19562608 TI - Associations between objectively measured habitual physical activity and adiposity in children and adolescents: Systematic review. AB - This review examined recent evidence on associations between objectively measured habitual physical activity and adiposity. A search for observational studies was carried out using several electronic databases from June 2004-June 2008. Of 1 255 potentially eligible papers, 47 papers were included, which described 48 studies. Most studies (41/48; 85%) were cross-sectional and 31/48 (65%) used proxies for adiposity, such as body mass index (BMI) or BMI z-score as the outcome measure. Few studies (10%; 5/48) focused on pre-school children. There was consistent evidence of negative associations between objectively measured physical activity and adiposity: significant negative associations were observed in 38/48 (79%) of studies overall. The present review supports the hypothesis that higher levels of habitual physical activity are protective against child and adolescent obesity. However, prospective longitudinal studies are warranted; there is a need for more research on younger children, and for more 'dose-response' evidence. PMID- 19562609 TI - Do children perceive features of real and cartoon faces in the same way? Evidence from typical development and autism. AB - In the current study, typically developing children and children with autism spectrum disorders (ASD) were presented with a facial-feature discrimination task including both real and cartoon faces, displayed either upright or inverted. Results demonstrated that typically developing children were more accurate at discriminating facial features from upright than from inverted faces and that this effect was specific to real faces. By contrast, children with ASD failed to show such a specific pattern of performance for processing facial features displayed in real faces. Findings of the current study suggest that face type (real vs. cartoon) does not affect perceptual ability in children with ASD as it does in typically developing children. PMID- 19562610 TI - Non-bullous lesions as the first manifestation of bullous pemphigoid: a retrospective analysis of 24 cases. AB - OBJECTIVE: To explore the clinical features, laboratory findings and therapy of the patients with bullous pemphigoid initially manifested as non-bullous lesions. METHODS: The medical records of 24 cases of bullous pemphigoid initially manifested as non-bullous lesions were retrospectively analyzed. RESULTS: The male-female ratio was 2:1. The mean age of first onset was 61.71 +/- 15.82 years old. The patients presented with erythema, papules, papulovesicles, plaques, wheals, nodules or erythema multiform-like lesions before typical bullae appeared. The majority of the initial lesions were concurrent erythema, papules and plaques (11/24, 45.83%). The first blisters were located on the extremities in eight patients (33.33%). A limitation of the study was not being able to analyze the histopathological and immunopathological features of the initial non bullous lesions as it was a retrospective study. CONCLUSIONS: Nearly one-third of patients with bullous pemphigoid presented with multiform initial lesions, mainly erythema, papules and plaques simultaneously before the first blisters appeared. The extremities are the favorite sites of the first blisters. The diagnosis was confirmed by histopathology and immunofluorescence. PMID- 19562612 TI - Elevated cancer mortality in a German cohort of bitumen workers: extended follow up through 2004. AB - The mortality follow-up in a cohort of German asphalt workers covered an additional 6 years. Male workers (N = 7919) were classified into four exposure categories: (1) exposure to bitumen only, (2) to bitumen and coal tar, (3) neither to tar nor to bitumen, and (4) unknown exposure. Exposure-specific standardized mortality ratios (SMR) and associated 95% confidence intervals (CI) based on age- and calendar period-specific national mortality rates were calculated. To compare exposed and unexposed workers, relative risks were estimated by Poisson regression. By the end of 2004, 835 workers had died. The SMR for lung cancer was 1.77 (95% CI 1.46-2.16). Head and neck cancer showed an SMR of 2.36 (95% CI 1.78-3.07). Bladder cancer mortality was elevated threefold. Significantly elevated cancer-related SMRs were also found for all malignant tumors. In addition, elevated mortality rates of nonmalignant causes such as alcoholism, liver cirrhosis, and unnatural causes of deaths including accidents were observed. After stratification by exposure group, cancer mortality was elevated among bitumen-exposed and unexposed subjects. In the internal analysis, the association between lung cancer and bitumen exposure was weakened as compared with the previous follow-up (relative risk [RR] = 1.15; 95% CI 0.72-1.84). The follow-up demonstrated an excess of cancer in this cohort of asphalt workers. However, the observed mortality patterns were not clear. Although exposure to bitumen cannot be ruled out as being responsible for the observed results, a higher prevalence of alcohol and tobacco consumption may partially explain the observed risk increases. Exposure assessment in future studies should account for multiple occupational agents and nonoccupational factors to rule out that the observed differences in SMR are not due simply to random variation. PMID- 19562611 TI - Role of metabolic syndrome components in human immunodeficiency virus-associated stroke. AB - Metabolic syndrome (MetS) is a cluster of risk factors, including elevated mean arterial pressure (MAP), atherogenic dyslipidemia (elevated triglycerides [TRG]), abdominal obesity (increased body mass index [BMI]), glucose intolerance (elevated glucose [GLU]), and prothrombotic/inflammatory state (increases in uric acid [UA]), that are associated with increased risk of cerebrovascular disease. We studied if an association existed between MetS components and human immunodeficiency virus (HIV)-associated cryptogenic strokes-those not caused by HIV complications, endocarditis, or stimulant abuse. We performed a retrospective case-control study. Eleven cryptogenic strokes were identified from 2346 HIV infected (HIV+) participants. Each case was matched by age, sex, and date of stroke diagnosis to five HIV+ controls without stroke. Nonparametric stratified Wilcoxon ranked sum tests with subsequent mixed effect logistic regression determined the influence of each MetS component on HIV-associated cryptogenic stroke. Although each MetS component appeared higher for HIV+ cases with cryptogenic strokes than HIV+ controls, only MAP (odds ratio [OR] = 5.70, 95% confidence interval [CI] = 1.15-28.3) and UA (OR = 1.88, 95% CI = 1.06-3.32) were statistically different. A significantly higher percentage of HIV-associated cryptogenic stroke cases met criteria for MetS (4/11 = 36%) compared to HIV+ controls (6/55 = 11%). This observational study suggests a possible role for MetS components in HIV+ cryptogenic stroke cases. Although MetS is defined as a constellation of disorders, elevated hypertension and hyperuricemia may be involved in stroke pathogenesis. Reducing MetS component levels in HIV+ patients could therefore protect them from subsequent stroke. PMID- 19562613 TI - A rare case of non-Hodgkin lymphoma in a pacemaker pocket. PMID- 19562614 TI - pY-STAT3 and p53 expression predict outcome for poor prognosis diffuse large B cell lymphoma treated with high dose chemotherapy and autologous stem cell transplantation. AB - The purpose of this study was to evaluate the ability of biomarkers to predict 5 year event-free survival (EFS) of poor prognosis patients with diffuse large B cell lymphoma (DLBCL) who were treated on a prospective clinical trial with upfront high dose chemotherapy (HDCT) and autologous stem cell transplantation (ASCT). We previously reported 51 patients with DLBCL treated with one cycle each of cyclophosphamide, doxorubicin, vincristine, prednisone (CHOP), dose-intensive cyclophosphamide, etoposide, cisplatin (DICEP), and carmustine, etoposide, Ara-C, and melphalan (BEAM)/ASCT. Of these patients, 33 had DLBCL and suitable tissue for immunohistochemical (IHC) biomarker evaluation. We found no statistically significant association between EFS and age adjusted International Prognostic Index (IPI) score, bulk over 10 cm, germinal center B-cell phenotype, or expression of BCL-2 or BCL-6 biomarkers. However, the detection of pY-STAT3 expression was associated with improved 5-year EFS (93%versus 47%, p = 0.006), and p53 expression was associated with lower 5-year EFS (47%versus 83%, p = 0.025). Predictive ability was improved by combining pY-STAT3 and p53 expression. Specifically, 5-year EFS rates were 93% for pY-STAT3+, 77% for pY-STAT3-/p53-, and 20% for pY-STAT3-/p53+ patients (p = 0.0002). In conclusion, pY-STAT3 and p53 expression may help predict outcome of HDCT for DLBCL, and further study of these biomarkers is warranted. PMID- 19562615 TI - Concurrent herpes simplex viral lymphadenitis and chronic lymphocytic leukemia/small lymphocytic lymphoma, mimicking large-cell (Richter) transformation. PMID- 19562617 TI - A fertile XY/XX chimeric male with chronic myeloid leukemia in a minor 46,XX cell line and a history of polycythemia vera and trisomy 9 in the major 46,XY cell line. PMID- 19562616 TI - Variable contribution of monoclonal antibodies to ADCC in patients with chronic lymphocytic leukemia. AB - Monoclonal antibodies (mAbs) are increasingly used in treatment protocols for chronic lymphocytic leukemia (CLL). Here we determined (i) the extent of antibody dependent cellular cytotoxicity (ADCC) of four different mAbs against primary CLL cells, (ii) whether ADCC correlates with antigen density on CLL cells, and (iii) whether allogeneic natural killer (NK) cells display superior ADCC than autologous. Effector cells for ADCC were (i) NK-92 cells not expressing FcR, (ii) NK-92 cells transfected with a high-affinity Fc receptor, (iii) autologous NK cells from patients with CLL, (iv) allogeneic NK cells. Results suggest that ADCC contributes to killing of CLL cells by anti-CD20 antibodies (rituximab and veltuzumab), whereas mAbs against CD22 (epratuzumab) and CD23 (lumiliximab) showed minimal ADCC. The magnitude of anti-CD20 mediated ADCC did not correlate with antigen density of CD20. ADCC was not influenced by the FcR genotype expressed by autologous NK cells. Allogeneic NK cells were superior to autologous NK cells in killing primary CLL cells. PMID- 19562618 TI - JAK2 V617F mutation is associated with 5q- syndrome in Chinese. AB - JAK2 V617F mutation is mostly seen in BCR-ABLI negative myeloproliferative neoplasms. Among other myeloid neoplasms, it occurs with remarkably high frequency in refractory anemia with ring sideroblasts associated with marked thrombocytosis, a group of myeloid neoplasms with both dysplastic and proliferative features. It has also been reported in occasional cases of myelodysplastic syndrome with isolated del(5q), often with a diagnosis of refractory cytopenia with multilineage dysplasia. We performed a retrospective analysis of JAK2 V617F mutation in Chinese patients with myeloid neoplasms and isolated del(5q), and were able to demonstrate the frequent occurrence of JAK2 V617F mutation in 5q- syndrome. PMID- 19562619 TI - Mantle cell lymphoma - does primary intensive immunochemotherapy improve overall survival for younger patients? AB - MCL is a rare entity of non-Hodgkin lymphoma, hitherto considered incurable. There is no standard therapy, but the current treatment results do seem to have led to a prolongation of the median survival from 3 to 5 years. Following CHOP like induction, high-dose radiochemotherapy, and autologous stem cell transplantation (ASCT) chemotherapy has been shown in a controlled trial to be superior in younger patients, but does not, however, lead to long-term freedom from disease. Results of recent prospective but uncontrolled trials of more intensive frontline immunochemotherapy containing cytarabine and rituximab followed by ASCT, however, now for the first time indicate plateaus of the curves of event-free, progression-free and overall survival, suggesting cure, but more studies and longer follow-up is needed. Following relapse, autologous stem-cell transplantation does not seem to be of value, but graft-versus-lymphoma effect has been documented, and allogeneic stem cell transplantation with reduced intensity conditioning is emerging as the treatment of choice in this setting. PMID- 19562620 TI - JAK2 V617F mutation in myeloid neoplasms with 3q21 and 3q26 abnormalities in Chinese patients. PMID- 19562621 TI - Ultra structure of camel platelets. PMID- 19562622 TI - To eat or not to eat? Effect of fasting prior to abdominal sonography examinations on the quality of imaging under routine conditions: A randomized, examiner-blinded trial. AB - OBJECTIVE: Although often recommended, it is unclear whether fasting enhances the imaging quality of abdominal sonography examinations. The aim of this study was to produce experimental evidence of the effect of fasting on the imaging quality of abdominal organs. MATERIAL AND METHODS: Formally consenting medical inpatients who underwent elective abdominal sonography examinations at a university medical center were randomized to either a fasting or a non-fasting preparation. Blinded examiners evaluated the imaging quality of 11 anatomical regions. The primary end point was the proportion of completely evaluable patients for each region. In secondary analyses, values of an imaging index reflecting the mean imaging quality of all regions (range 0-1) were compared. RESULTS. Of 280 screened patients, 102 (36%) met the exclusion criteria and 35 (13%) declined participation. Of the 143 randomized patients, 130 (91%) were included in the primary analyses (66 fasting, 64 non-fasting). The proportion of completely evaluable patients did not differ significantly for any of the 11 regions, but a large nominal difference occurred for the gallbladder (45/66 (73%) fasting versus 34/64 (56%) non-fasting patients, p=0.051). The median (range) imaging index was 0.57 (0.14-0.95) for fasting and 0.43 (0.00-1.00) for non-fasting subjects (p =0.078). A significant (p=0.002) difference favoring fasting was detected in the post-hoc subgroup analyses for male patients. CONCLUSIONS: For examinations of the gallbladder and for male patients, fasting might improve the sonographic imaging quality to some extent. Overall, no significant improvement in the imaging quality of abdominal organs was reached with a fasting preparation. PMID- 19562623 TI - Assessment of liver stiffness in patients after living donor liver transplantation by transient elastography. AB - OBJECTIVE: Recurrence of hepatitis and progression of fibrosis are major problems in liver transplantation (LT) for patients with hepatitis C. Liver stiffness measurement (LSM) by transient elastography correlates well with histologic liver fibrosis stages in chronic liver diseases. The aim of this study was to evaluate the usefulness of transient elastography for the assessment of fibrosis in patients after living donor LT. MATERIAL AND METHODS: Seventy-nine patients who visited our institution, and in whom LSM was successfully evaluated, were enrolled in the study. The patients were divided into three groups according to positivity for hepatitis C antibody and hepatitis B surface antigen as the hepatitis C virus (HCV) group (n=37), the hepatitis B virus (HBV) group (n=10), and the NBNC (negative for both hepatitis B and C) group (n=32). The correlation between LSM and histologic fibrosis stage was assessed in 36 patients. LSM was also compared with regard to the effect of interferon therapy in HCV patients. RESULTS: The median value for liver stiffness was 6.8 kPa and the median time from LT was 3.1 years. In patients who underwent liver biopsy, stiffness was significantly correlated with the stages of fibrosis (p<0.001, rho = 0.848). In patients who received interferon therapy after LT, the LSM decreased over time in those with a sustained virological response, whereas LSM increased in patients without a response. CONCLUSION: Transient elastography may be an appropriate non invasive procedure to sequentially assess the progression of liver fibrosis in patients after LT. PMID- 19562624 TI - Diagnostic yield of blood cultures from antibiotic-naive and antibiotically treated patients with haematological malignancies and high-risk neutropenia. AB - In patients with prolonged episodes of neutropenia, infections are associated with significant mortality. The exact diagnostic yield of blood cultures in this high-risk population is still unclear. To assess the yield of blood cultures, the spectrum of pathogenic organisms and the influence of blood culture results on the therapeutic management, we retrospectively evaluated the results from 2520 blood cultures obtained from 126 consecutive patients with high-risk neutropenia. Bacterial pathogens were detected in 219 blood culture samples (8.7%) of which 172 were Gram-positive and 47 were Gram-negative bacteria. Fungal pathogens were found in 13 blood cultures. A higher rate of Gram-positive pathogens and of fungi was found in patients with central venous catheters. Pathogens were detected in 14.3% of blood cultures obtained before the institution of antibiotic treatment and in 7% of blood cultures obtained under antibiotic treatment. Treatment was modified in 116/232 (50%) of positive blood culture findings. In patients with high-risk neutropenia, blood cultures are a valid diagnostic tool, both in antibiotic-naive patients and in patients receiving antibiotic treatment, and provide important information for clinical decision making. The epidemiological data obtained are helpful for selecting empirical antibiotic treatment regimens. PMID- 19562625 TI - Client perceptions of the Tree Theme Method™: a structured intervention based on storytelling and creative activities. AB - The Tree Theme Method (TTM) is an intervention based on sessions involving creative activities and life storytelling/story-making, in which the client paints trees representing various periods of his/her life. The aim of this study was to investigate clients' experiences of participating in a TTM intervention and their perceptions of the therapeutic relationship. Thematic interviews were undertaken. Twenty clients attending general outpatient mental healthcare units were recruited after having participated in the intervention. A qualitative content analysis resulted in six categories: "From feeling a pressure to perform to becoming focused and expressive", "Expressing oneself and one's life situation led to awakening of memories and feelings", "New perspectives of self-image, everyday life and relations to others", "Story-making led to shaping and reconstructing one's life story", "Interaction was of importance when reconstructing one's life story" and, finally, "The attitude of the occupational therapist was of importance for the development of the therapeutic relationship". There seemed to be a close association between the intervention and the therapeutic relationship in starting a process of opening up new perspectives on everyday life, but there is a need for further studies including therapists' experience of using the TTM and their perception of the client-therapist relationship. PMID- 19562626 TI - Contacts related to psychiatry and substance abuse in Norwegian casualty clinics. A cross-sectional study. AB - OBJECTIVE: To provide quantitative measurement and analysis of the frequency with which patients contact emergency primary healthcare services in Norway for psychiatric illness, including substance misuse. Characteristics of the patient group and their contact times were also addressed. DESIGN: Cross-sectional observational study. SETTING: Data were collected from one district-based and one city-based casualty clinic in Norway. SUBJECTS: Patients seeking medical care during the whole of 2006. MAIN OUTCOME MEASURES: Patients' diagnoses, age, gender, and time of contact. RESULTS: Diagnoses related to psychiatric illness were found in 2.7% of all events at the casualty clinics, but were relatively more frequent at night (5.6%) and for home visits and out-of-office emergency responses combined (8.4%). Prevalence was almost doubled during the July holiday month. Prevalence remained relatively constant between ages 15 and 59. The most frequently diagnosed subgroups were depression/suicidal behaviour, anxiety, and substance abuse (21.3%) of which 76.8% was alcohol-related. Gender and age differences within diagnostic subgroups were identified. For example, substance abuse was more prevalent for men, while anxiety was more prevalent for women. CONCLUSION: Psychiatric illness and substance misuse have relatively low presentation rates at Norwegian casualty clinics, compared with established daytime attendance at general practitioners. However, the prevalence increases during periods with lowered availability of primary and specialist psychiatric healthcare. These data have implications for the allocation of resources to patient treatment and provide a foundation for future research into provision of emergency healthcare services for this group of patients. PMID- 19562628 TI - The relation between race-related implicit associations and scalp-recorded neural activity evoked by faces from different races. AB - The neural correlates of the perception of faces from different races were investigated. White participants performed a gender identification task in which Asian, Black, and White faces were presented while event-related potentials (ERPs) were recorded. Participants also completed an implicit association task for Black (IAT-Black) and Asian (IAT-Asian) faces. ERPs evoked by Black and White faces differed, with Black faces evoking a larger positive ERP that peaked at 168 ms over the frontal scalp, and White faces evoking a larger negative ERP that peaked at 244 ms. These Black/White ERP differences significantly correlated with participants' scores on the IAT-Black. ERPs also differentiated White from Asian faces and a significant correlation was obtained between the White-Asian ERP difference waves at approximately 500 ms and the IAT-Asian. A positive ERP at 116 ms over occipital scalp differentiated all three races, but was not correlated with either IAT. In addition, a late positive component (around 592 ms) was greater for the same race compared to either other race faces, suggesting potentially more extended or deeper processing of the same race faces. Taken together, the ERP/IAT correlations observed for both other races indicate the influence of a race-sensitive evaluative process that may include early more automatic and/or implicit processes and relatively later more controlled processes. PMID- 19562629 TI - Individualism, conservatism, and radicalism as criteria for processing political beliefs: a parametric fMRI study. AB - Politics is a manifestation of the uniquely human ability to debate, decide, and reach consensus on decisions affecting large groups over long durations of time. Recent neuroimaging studies on politics have focused on the association between brain regions and specific political behaviors by adopting party or ideological affiliation as a criterion to classify either experimental stimuli or subjects. However, it is unlikely that complex political beliefs (i.e., "the government should protect freedom of speech") are evaluated only on a liberal-to conservative criterion. Here we used multidimensional scaling and parametric functional magnetic resonance imaging to identify which criteria/dimensions people use to structure complex political beliefs and which brain regions are concurrently activated. We found that three independent dimensions explained the variability of a set of statements expressing political beliefs and that each dimension was reflected in a distinctive pattern of neural activation: individualism (medial prefrontal cortex and temporoparietal junction), conservatism (dorsolateral prefrontal cortex), and radicalism (ventral striatum and posterior cingulate). The structures we identified are also known to be important in self-other processing, social decision-making in ambivalent situations, and reward prediction. Our results extend current knowledge on the neural correlates of the structure of political beliefs, a fundamental aspect of the human ability to coalesce into social entities. PMID- 19562630 TI - Infants' neural processing of positive emotion and eye gaze. AB - Previous research demonstrated that young infants' neural processing of novel objects is enhanced by a fearful face gazing toward the object. The current event related potential (ERP) study addresses the question of whether this effect is driven by the particular threat-value of a fearful expression or whether a positive emotion could elicit a similar response. Three-month-old infants' brain responses were measured while infants were presented with happy and neutral faces gazing toward simultaneously presented objects (Experiment 1) or happy and neutral faces gazing away from objects (Experiment 2). Then the objects were presented again without the face. While infants showed an increased neural response for happy relative to neutral faces looking towards objects, infants did not differentiate between happy and neutral faces gazing away from the objects. Furthermore, infants showed no different response to objects alone in Experiment 1. However, infants responded with an increased negative central component (Nc) indicating increased attention for objects in the neutral face condition in Experiment 2. The current results confirm previous findings showing that infants allocate increased attention to an emotional face if it directs eye gaze toward an object in the environment. However, a happy expression does not increase subsequent processing of the gaze-cued object. The findings are discussed in terms of early social cognitive development. PMID- 19562631 TI - Decreased ventral anterior cingulate cortex activity is associated with reduced social pain during emotional support. AB - People feel psychological pain when they are excluded, and this pain is often attenuated when emotional support is received. It is therefore likely that a specific neural mechanism underlies the detection of social exclusion. Similarly, specific neural mechanisms may underlie the beneficial effects of emotional support. Although neuroimaging researchers have recently examined the neural basis of social pain, there is presently no agreement as to which part of the anterior cingulate cortex (ACC) is involved in the perception and modulation of social pain. We hypothesized that activity in those brain regions that are associated with social pain would be correlated with decrements in social pain induced by emotional support. To examine the effects of emotional support on social pain caused by exclusion, we conducted an fMRI study in which participants played a virtual ball-tossing game. Participants were initially included and later excluded from the game. In the latter half of the session from which participants were excluded, participants received emotionally supportive text messages. We found that emotional support led to increased activity in the left lateral/medial prefrontal cortices and some temporal regions. Those individuals who experienced greater attenuation of social pain exhibited lower ventral ACC and higher left lateral prefrontal cortex activation. These results suggest that the ventral ACC underlies social pain, and that emotional support enhances prefrontal cortex activity, which in turn may lead to a weakened affective response. PMID- 19562632 TI - Increased oxytocin concentrations and prosocial feelings in humans after ecstasy (3,4-methylenedioxymethamphetamine) administration. AB - MDMA (3,4-methylenedioxymethamphetamine or "ecstasy") is a recreationally used drug with remarkable and characteristic prosocial effects. In spite of abundant attention in the scientific literature, the mechanism of its prosocial effects has not been elucidated in humans. Recently, research in animals has suggested that the neuropeptide oxytocin may induce these effects. In a double blind, randomized, crossover, and placebo-controlled study in 15 healthy volunteers we assessed blood oxytocin and MDMA concentrations and subjective prosocial effects after oral administration of 100 mg MDMA or placebo. MDMA induced a robust increase of blood oxytocin concentrations and an increase of subjective prosocial feelings. Within subjects, the variations in these feelings were significantly and positively correlated with variation in oxytocin levels, and the correlations between these feelings and oxytocin were significantly stronger than those between these feelings and blood MDMA levels. MDMA induces oxytocin release in humans, which may be involved in the characteristic prosocial effects of ecstasy. PMID- 19562634 TI - Sonographic features of anorectal atresia at 12 weeks. AB - We present the sonographic features of a fetus with anal atresia at 12 weeks of gestation. Follow-up ultrasound examination at 17 week revealed apparently normal bowel. Spontaneous miscarriage occurred at 18 weeks and postmortem examination showed anorectal atresia and arthrogryposis multiplex. It seems that dilatation of the bowel in the early pregnancy is a possible marker for anorectal atresia, and the abnormality may be overlooked if a mid-trimester scan alone is performed. PMID- 19562633 TI - Factors associated with phosphodiesterase type 5 inhibitor treatment satisfactions: results of patient interrogation. AB - INTRODUCTION: Phosphodiesterase type 5 (PDE5) inhibitor therapy is an efficacious means of treatment for erectile dysfunction (ED). PDE5 inhibitors supply penile erection by inhibiting the hydrolysis of cGMP and therefore relaxing the corpus cavernosum. In this study, retrospective evaluation of those patients who were admitted to our clinic with the complaint of ED and who were recommended on PDE5 inhibitor treatment in terms of follow-up results and patient satisfaction were aimed. METHOD: The patients were called by phone and after informing about the study and taking the informed consent, patient satisfaction with the treatment, purposes of withdrawal, treatment alterations and partner satisfaction were investigated. RESULTS: Interviews were made with 345 patients, who accepted to enroll in the study and the mean patient age was 56 +/- 11.2 years. Of the patients 66.4% were learned to be satisfied with the treatment. It was determined that 10.7% of the patients have never used the medication and 50% could not continue because of high drug cost. It was recognised that 50.2% of the patients who are not satisfied with the treatment tried another PDE5 inhibitor. The success rate of the treatment was found to be higher in the followed-up group than those losses to follow-up. CONCLUSION: Therapy with PDE5 inhibitors is an effective means of ED treatment. The importance of doctor-patient communication should be considered, and the patient should be advised for adaptation to follow up program. High drug cost is a significant predictor of patient compliance to treatment continuation. PMID- 19562635 TI - Good perinatal outcome of the pump twin in the twin reversed-arterial- perfusion sequence diagnosed at the ninth week of pregnancy and managed conservatively. PMID- 19562637 TI - Monochorionic dizygotic twins in a spontaneous pregnancy: a rare case report. AB - Traditionally, monochorionicity in multiple pregnancies is associated with monozygocity. We present a case of a spontaneous, monochorionic dizygotic, sex discordant twin pregnancy. The diagnosis of monochorionicity was initially done during first-trimester ultrasound evaluation and then confirmed by postnatal placental pathology. Furthermore, both twins were found to have blood chimerism. We also review the literature on dizygotic-monochorionic twins and blood chimerism. We suggest that further prospective postnatal genetic studies are needed to define the reliability of prenatal diagnosis of identical twins in cases of monochorionicity. PMID- 19562638 TI - SIL-TAL1 fusion gene negative impact in T-cell acute lymphoblastic leukemia outcome. AB - SIL-TAL1 fusion gene and the ectopic expression of HOX11L2 are common molecular abnormalities in T-cell acute lymphoblastic leukemia (T-ALL). To verify their influence on outcome, we analyzed a Brazilian pediatric T-ALL series of cases. One hundred and ninety two children, age ranged 0-21 years old, were consecutively diagnosed and treated. Reverse transcriptase-polymerase chain reaction (RT-PCR) technique was used to identify the molecular alterations. Kaplan-Meyer method was applied to estimate overall survival. The most frequent maturation stage was T-IV (40.1%), and 30.7% of cases were CD10(+). SIL-TAL1(+) and HOX11L2(+) accounted for 26.7% and 10.3% of the cases, respectively. The overall survival (OS) was 74% in 80-month follow-up. HOX11L2(+) was not predictive factor for outcome. Considering patients younger than nine years-old, those with SIL-TAL1(+) presented a poorer outcome (p = 0.02). The results of this study suggest that in the Brazilian population only the presence of SIL-TAL1 can predict outcome in a restricted group of patients. PMID- 19562639 TI - Autologous and allogeneic stem cell transplantation: rising therapeutic promise for mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is currently an incurable neoplasm with a median survival duration of 3-5 years. The clinical results of therapy with cyclophosphamide, doxorubicin, vincristine, and prednisone or similar regimens are inadequate,leading to widespread exploration of the use of autologous stem cell transplantation (ASCT) during the first remission. In the pre-rituximab era, early ASCT extended the median remission duration by 1-2 years, but most patients eventually experienced relapse. With the advent of rituximab and its incorporation into stem cell mobilization and conditioning regimens, several research groups have reported improved outcomes, including the emergence of early survival curve plateaus that suggest a cured fraction. Intensive chemoimmunotherapy with rituximab and hyperfractionated cyclophosphamide,vincristine, doxorubicin, and dexamethasone has been reported to have similarly favorable results. Therefore, the addition of rituximab to intensive chemotherapy or ASCT regimens may be curative in patients undergoing frontline treatment for MCL. In the relapsed or refractory disease setting, the clinical results of ASCT remain inadequate. However, the increasing safety and high efficacy of non-myeloablative stem cell transplantation (SCT) suggests that it is the most appropriate transplantation modality in patients with relapsed or refractory MCL when a suitable donor is available. PMID- 19562640 TI - Personalized medicine: a paradigm for a sustainable pharmaceutical industry? AB - Personalized medicine is a custom-tailored approach to patient treatment based on individual genetic traits. In personalized medicine, a patient group is characterized by a clinical biomarker that has been correlated to a differential response to drug treatment. During the past decade, several developments in the understanding of the structure and function of the human genome have occurred that bring personalized medicine closer to becoming a reality. The promise of personalized medicine lies in a clinical biomarker-driven patient stratification, and focused smaller-sized clinical trials that result in a shorter development time and reduced overall development cost. Personalized medicine has the potential to offer a new business model for the pharmaceutical industry by providing a more efficient drug discovery process with reduced cost. PMID- 19562641 TI - Recent progress in the discovery and development of negative allosteric modulators of mGluR5. AB - Negative allosteric modulators (NAMs) of metabotropic glutamate receptor subtype 5 (mGluR5) have attracted significant interest in the pharmaceutical industry as potential therapies for a variety of diseases of the CNS. Well-characterized small-molecule mGluR5 NAMs have demonstrated efficacy in several preclinical models of disease, and examples of the clinical efficacy of such NAMs have been reported in recent years. Advances, both in the discovery and development of new small molecules and in the clinical evaluation of known compounds, have been significant in the past 2 years. Recent progress in preclinical drug discovery efforts has used multiple strategies for the discovery of new chemical scaffolds, including the structural modification of known compounds, the rational design of new chemical scaffolds based on known compounds, and the development of screening approaches for new, structurally distinct scaffolds. Progress in the clinical setting with mGluR5 NAMs has also resulted in important recent advances, including the completion of proof-of-concept studies. PMID- 19562642 TI - PDE10A inhibitors: an assessment of the current CNS drug discovery landscape. AB - PDE10A is a dual substrate PDE that is highly expressed in medium spiny neurons of the striatal complex. The inhibition of PDE10A produces effects that modulate basal ganglia function in ways that suggest a particular therapeutic utility in the treatment of psychosis in schizophrenia. Significant understanding of PDE10A at the molecular level has helped to guide efforts in inhibitor design, and many different inhibitor classes have now been discovered. At least one PDE10A inhibitor has been advanced into clinical trials to begin to test the hypothesis that such agents may be useful in the treatment of psychosis. PMID- 19562643 TI - Glycine transport inhibitors for the treatment of schizophrenia: symptom and disease modification. AB - Schizophrenia is a severe neuropsychiatric disorder for which there is no adequate current treatment. Recent theories about the molecular basis of schizophrenia focus on disturbances of glutamatergic neurotransmission, particularly at NMDA-type glutamate receptors (NMDARs). NMDARs are regulated in vivo by the amino acids glycine and D-serine. Glycine levels, in turn, are regulated by glycine transporter type 1 (GlyT1), which serves to maintain low subsaturating glycine levels in the vicinity of the NMDAR. Therefore, one proposed approach to the treatment of schizophrenia is via the inhibition of GlyT1-mediated transport. During the past decade, several well-tolerated, high affinity glycine transport inhibitors (GTIs) have been developed that demonstrate the ability to potentiate NMDAR-mediated neurotransmission in animal models relevant to schizophrenia. In addition, clinical trials have been conducted with sarcosine (N-methylglycine), a naturally occurring GTI. Issues related to clinical proof-of-concept studies with high-affinity GTIs in schizophrenia are discussed in this review. PMID- 19562644 TI - Prospects for the development of fusion inhibitors to treat human respiratory syncytial virus infection. AB - Human respiratory syncytial virus (hRSV) is a significant cause of respiratory illness in at-risk pediatric patients, immunocompromised adults and the elderly. No vaccine is currently available for the virus and treatment options are limited to the prophylactic treatment of at-risk infants with the mAb palivizumab (Synagis) and to therapeutic intervention with the nucleoside analog ribavirin (Rebetol). The clinical use of these agents is limited and a need exists for more effective treatment for the at-risk population. The merging of viral and cellular membranes is a crucial event in the hRSV life cycle that enables the virus to enter a host cell. The multistep fusion process is facilitated by the substantial refolding of a trimeric class I fusion protein (F protein), which is the main target of fusion inhibitors. Several small-molecule fusion inhibitors have been discovered, of which some have progressed significantly in the drug development process. BTA-9881 (Biota Holdings Ltd/MedImmune) and TMC-353121 (Johnson & Johnson) are the most advanced of this drug class. In addition, progress has been made in the development of next-generation antibodies such as motavizumab (Numax; MedImmune). This review will discuss the status and latest developments of compounds and antibodies that inhibit hRSV fusion. PMID- 19562645 TI - The importance of drug-target residence time. AB - The importance of kinetics in drug-target interactions, and particularly the residence time of a drug with its target, is increasingly recognized to play a pivotal role in determining both the efficacy and toxicity of a drug. Drug residence time can often be demonstrated to be a key differentiating factor between drugs that act upon a common target. Drug-target residence time can result in either favorable or unfavorable outcomes, and the use of such information could lead to the more efficient design of best-in-class drugs. This review highlights several key concepts and observations related to drug-target residence time, and suggests the use of a kinetics-perceptive and energetics informed approach to address the challenges facing current drug discovery efforts. PMID- 19562646 TI - Direct Factor Xa and direct thrombin inhibitors: a clinical trial update. AB - Anticoagulant use is recommended for the prevention and treatment of several thromboembolic disorders including venous thromboembolism. However, anticoagulants are often underprescribed because of the disadvantages associated with conventional agents, including the requirement for routine clinical monitoring and parenteral administration. Recent research has focused on the development of agents that target specific factors in the coagulation cascade, primarily Factor (F)Xa and thrombin. There are several new, targeted oral anticoagulants in development that could potentially provide more effective and more convenient anticoagulation than conventional agents. This review provides an update on clinical trials of the direct FXa and thrombin inhibitors. PMID- 19562647 TI - Physiologically based pharmacokinetic (PBPK) modeling and simulation: applications in lead optimization. AB - Physiologically based pharmacokinetics (PBPK) models are increasingly being used in the lead optimization (LO) process. Although there are currently few literature reports of the application of PBPK, the scope of PBPK modeling is expanding and there is a steady increase in the number of publications in this field. Recent publications covering four important areas of the application of PBPK modeling in LO have been reviewed. PMID- 19562648 TI - GPR119 agonists: a promising new approach for the treatment of type 2 diabetes and related metabolic disorders. AB - Type 2 diabetes (T2D) and associated obesity have reached epidemic proportions, and there is an increasing need for orally effective agents that regulate glucose homeostasis with a concurrent reduction in body weight. GPR119, a class-A (rhodopsin-like) G protein-coupled receptor, expressed primarily in the human pancreas and gastrointestinal tract, has attracted considerable interest as a T2D drug target in the last three to five years. The activation of GPR119 increases the intracellular accumulation of cAMP, leading to enhanced glucose-dependent insulin secretion and increased levels of the incretin hormones GLP-1 (glucagon like peptide 1) and GIP (glucose-dependent insulinotropic peptide). In rodent models, orally available GPR119-specific agonists have been shown to attenuate blood glucose levels with a simultaneous body weight loss. This review summarizes the research leading to the identification of GPR119 as a potential drug target for T2D and related metabolic disorders. In addition, an overview of the recent progress made in the discovery of orally active GPR119 agonists is provided. PMID- 19562649 TI - Is there a future for Aurora kinase inhibitors for anticancer therapy? AB - The development of Aurora kinase inhibitors is a competitive research field, with many inhibitors currently being evaluated in preclinical and clinical studies. Progress during the past few years, both preclinically and clinically, has increased the evidence supporting Aurora kinases as promising molecular targets for the treatment of cancer. Aurora kinase inhibitors differ based on their selectivity within the Aurora kinase family and their cross-reactivities with other kinases. Additional factors that will contribute to the success or failure of the Aurora kinase inhibitors include: routes of administration, drug-like properties, workable combinations with approved drugs, adequate clinical development paths, and the identification of the appropriate patient population. The clinical trial results that are emerging for the most advanced inhibitors are promising, and it is probable that clinical proof of concept will be achieved, and that Aurora kinase inhibitors will be part of treatment for cancer in the future. PMID- 19562650 TI - Small molecules targeting sodium and calcium channels for neuropathic pain. AB - Neuropathic pain is a chronic disease, which impacts millions of individuals worldwide. The condition is currently treated with several drugs that provide pain relief that is inconsistent and complicated by CNS or cardiovascular (CV) side effects. Voltage-gated sodium channels (VGSCs) and voltage-gated calcium channels (VGCCs) are of particular interest as targets for neuropathic pain because they control electrical signals in both the central and peripheral nervous system. Recent research has demonstrated that the expression of voltage gated ion channels changes significantly under conditions of neuropathic pain in rodents and humans. Selective modulation of the channels involved in the pathology of the disease, while sparing the channels that are essential for normal nociception, offers promising opportunities for therapeutic intervention. This review summarizes recent developments of small molecules that target VGSCs and VGCCs. PMID- 19562652 TI - Comparison of urinary cytokines after ingestion of cranberry juice cocktail in pregnant subjects: a pilot study. AB - Our objective was to evaluate urinary cytokine excretion after daily cranberry or placebo exposure in pregnant women. Four-hour urine samples were collected from 27 pregnant women subjects who were randomized to cranberry juice cocktail or placebo in three treatment arms: A: Cranberry (C) two times daily (C, C; n = 10 pregnant); B: cranberry in the AM, then placebo (P) in the PM (C, P; n = 9 pregnant); and C: placebo two times daily (P, P; n = 8 pregnant). Urinary cytokines were measured using commercially available kits. There was a statistically significant difference in interleukin (IL)-6 of the urinary cytokines between the multiple daily cranberry dosing group (group A [C, C]): median, 3.16 (range, 0.01 to 7.34) and the placebo group (group C [P, P]): 9.32 (0.53 to 29.61 pg/mL; p = 0.038, Kruskal-Wallis test). We concluded that a difference in IL-6 was found in the multiple daily cranberry dosing groups compared with placebo. Lack of differences based on treatment allocation in the other cytokines may be due to beta error. Further studies are planned to evaluate these assays for the assessment of clinical effect. PMID- 19562653 TI - The effect of excess weight gain in teenage pregnancies. AB - Excess weight gain in pregnancy, as defined by the Institute of Medicine (IOM), has been linked to adverse obstetrical outcomes. However, this relationship has not been examined in the younger maternal population. Our aim was to study excess weight gain in our inner-city teenage population. In this retrospective cohort study, we reviewed all nulliparous teenage deliveries between 2000 and 2004. The groups were divided by IOM criteria into "underweight" (body mass index [BMI] <20 kg/m(2); n = 58), "normal" (BMI, 20 to 26.0 kg/m(2); n = 255), "overweight" (BMI, 26.1 to 29.0 kg/m(2); n = 54), and "obese" (BMI > 29.0 kg/m(2); n = 89). The groups were then compared according to normal (control, n = 257) and excess weight gain (n = 199). Frequencies and odds ratios (ORs) for adverse outcomes were calculated. Excess weight gain was associated with an increased risk for cesarean delivery (OR 1.96, 95% confidence interval [CI], 1.28 to 3.01) and postpartum fever (OR, 2.46; 95% CI, 1.13 to 5.35). Significant neonatal findings included higher birthweight (3199 g versus 2864 g; p < 0.0001) and increased risk of macrosomia (OR, 8.18; 95% CI, 2.02 to 32.99) in the excess weight gain group. We concluded that excess weight gain places teen mothers at increased risk for cesarean delivery, postpartum febrile morbidity, and macrosomia. Interventions aimed at optimal weight gain in teen pregnancies are warranted. PMID- 19562654 TI - [Short and long term results of endolymphatic sac surgery: a patient questionnaire based study]. AB - INTRODUCTION: The endolymphatic sac surgery for the treatment of Meniere's disease has been described since the 1920s. The success rate of this technique in terms of vertigo control has been reported to be 50-80%. However, the value of this treatment method remained controversial. Furthermore, the reliable identification of the endolymphatic sac intraoperatively can be challenging in some cases. This study examines the short-, middle- and long-term results in a larger cohort of patients. MATERIALS AND METHODS: In 74 patients, vertigo control, tinnitus and degree of satisfaction was evaluated by means of a questionnaire retrospectively. Additionally, the diagnostic value of the electrocochleography (EcochG) was determined. RESULTS: The overall vertigo control rate was more than 70% in patients followed up for two years and has reached 81% in patients followed up for more than two years. Hearing preservation rate was 61%. Tinnitus has disappeared in 11% and improved in 23% of the patients. In 47% of the patients it was unchanged and in 19% worsened. The difference in EcochG results pre- versus postoperative was highly significant. CONCLUSIONS: ELSS is a useful tool in the management of Meniere's disease, in particular in patients that do not benefit sufficiently from conservative therapy. PMID- 19562656 TI - [Self-desensitization with aspirin in a patient with aspirin intolerance]. PMID- 19562655 TI - [Nasal bone fracture: etiology, diagnostics, treatment and complications]. AB - BACKGROUND: The aim of the present evaluation was to analyse a large patient population with nasal bone fractures leading to surgical reposition of the nasal bone. Special attention was paid to age summit, the distribution of the seasons, the rate of open compared to closed nasal bone fractures, accompanying injuries, diagnosis and therapy as well as to the postoperative course. METHODS: A total of 300 patients was evaluated retrospectively. All patients had been treated between 1999 and 2004 regarding simple and complex nasal bone fractures. Data were analysed with regard to history, age, gender, diagnosis, therapy, results, and complications. RESULTS: The average age of the patients was 29.6+/-15.6 years at the time of fracture with clear predominance of the male gender (77%). Another age summit could be observed in patients of >60 years. The main reasons for nasal bone fracture were falls (30%), in elderly patients often caused by cardiac syncopes, and rows (28%). The most frequent findings were deviation of the longitudinal axis of the nose (59%) and traumatic deviation of the nasal septum (50%). 65 patients (22%) had open nasal bone fractures, in 5% of the cases septal hematoma could be observed. CONCLUSION: Closed reposition of the nasal bone is the therapy of choice in uncomplicated nasal bone fractures. Special attention has to be paid to the group of patients aged 60 years and older who often suffer from treatment requiring comorbidities. In these cases inpatient treatment must be considered. PMID- 19562657 TI - [Life-threatening epistaxis as the first symptom of an arteriovenous malformation]. PMID- 19562658 TI - Characterization and immunolocalization of arabinogalactan-proteins in roots of Echinacea purpurea. AB - From the high molecular weight fraction of an aqueous extract from roots of Echinacea purpurea L. Moench, arabinogalactan-proteins (AGPs), a class of proteoglycans proposed to be involved in cell differentiation and plant growth, were purified and characterized with regard to amino acid composition and structure of the polysaccharide moiety. The protein content of the AGP was 5.0 % (w/w) with the dominating amino acids Glx, Hyp, Asx, Ser, Thr and Ala. The highly branched polysaccharide moiety shows a linkage composition typical of AGPs with 1,3-, 1,6- and 1,3,6-linked galactopyranosyl residues and arabinofuranosyl residues predominantly as terminal and 1,5-linked residues. Terminal units of glucuronopyranose acid were also detected. Furthermore, a new method for the localization of AGPs in plant tissue has been developed. The synthetic (beta- D Glc)(3) Yariv phenylgycoside (betaGlcY) is known to specifically bind to AGPs. For immunolocalization, polyclonal betaGlcY-antibodies have been generated and were used to label Yariv-treated thin sections of roots from E. purpurea. After addition of the FITC-conjugated secondary antibody, the sections were analyzed by confocal laser scanning microscopy. AGPs are detected mainly in the central cylinder in the area of the xylem. Cell walls of vessels and tracheids are strongly labelled, especially at the inner area of the wall. Furthermore, there is intense labelling of the pit canals. PMID- 19562660 TI - Non-word repetition and literacy in Dutch children at-risk of dyslexia and children with SLI: results of the follow-up study. AB - This study related the non-word repetition (NWR) abilities of 4-year-old children at-risk of dyslexia and children with specific language impairment (SLI) to their reading abilities at age eight. The results show that the SLI group obtained the lowest NWR score and the at-risk group performed in-between the control and SLI group. Approximately half of the at-risk and SLI group showed reading difficulties. Literacy and NWR abilities were correlated for the at-risk group, but not for the SLI group. The findings point toward differences between the groups and suggest that dyslexia and SLI should not be treated as a similar disorder. PMID- 19562659 TI - Antiproliferative activity of the diterpenes jatrophone and jatropholone and their derivatives. AB - The antiproliferative activity of the diterpenes jatropholone A and B, 16 semi synthetic derivatives thereof, and that of jatrophone and its three derivatives was assessed on human cell cultures. The cells used comprised normal lung fibroblasts (MRC-5), gastric adenocarcinoma (AGS), leukemia (HL-60), lung cancer (SK-MES-1), and bladder carcinoma (J82). Jatropholone A ( 1) was inactive against all the tumor cell lines; however, its acetylation rendered a compound with antiproliferative activity. The epimeric jatropholone B ( 8) was active against all the cancer cell lines, and its derivatives presented different effects on the selected cell lines. While jatrophone ( 19) showed strong anticancer activity, its derivatives 9beta,13alpha-dihydroxyisabellione and 13alpha-hydroxy-9 beta acetoxyisabellione were less active. PMID- 19562661 TI - Can solving of wordchains be explained by phonological skills alone? AB - The present study focussed on the determinants for effective solving of the Wordchains Test (WCT) in a normative sample of Norwegian junior high-school students. Forty voluntary participants from a rural school district in Western Norway completed the WCT along with tests of general intellectual capacity, single word and non-word reading, auditory working memory, and visual scanning. All measures correlated significantly with each other except for general non verbal abilities were not correlated with visual scanning. A stepwise multiple regression analysis, using the WCT as the dependent variable, yielded a model that included single word reading, letter recognition, and working memory as independent variables. This model accounted for 75% of the variance in WCT performance. This finding suggests that phonological skills only have an indirect influence on WCT performance. Thus, the core deficit in dyslexia, i.e. impaired phonological skills, may be related to the development of word recognition skills, but have no direct effect on the WCT performance in a normative sample. PMID- 19562662 TI - Cross-resistance, inheritance and biochemical mechanisms of imidacloprid resistance in B-biotype Bemisia tabaci. AB - BACKGROUND: The B-type Bemisia tabaci (Gennadius) has become established in many regions in China, and neonicotinoids are extensively used to control this pest. Imidacloprid resistance in a laboratory-selected strain of B-type B. tabaci was characterised in order to provide the basis for recommending resistance management tactics. RESULTS: The NJ-Imi strain of B-type B. tabaci was selected from the NJ strain with imidacloprid for 30 generations. The NJ-Imi strain exhibited 490-fold resistance to imidacloprid, high levels of cross-resistance to three other neonicotinoids, low levels of cross-resistance to monosultap, cartap and spinosad, but no cross-resistance to abamectin and cypermethrin. Imidacloprid resistance in the NJ-Imi strain was autosomal and semi-dominant. It is shown that enhanced detoxification mediated by cytochrome-P450-dependent monooxygenases contributes to imidacloprid resistance to some extent in the NJ-Imi strain. Results from synergist bioassays and cross-resistance patterns indicated that target-site insensitivity may be involved in imidacloprid resistance in the NJ Imi strain of B. tabaci. CONCLUSION: Although oxidative detoxification mediated by P450 monooxygenases is involved in imidacloprid resistance in the NJ-Imi strain of B-type B. tabaci, target-site modification as an additional resistance mechanism cannot be ruled out. Considering the high risk of cross-resistance, neonicotinoids should be regarded as a single group when implementing an insecticide rotation scheme in B. tabaci control. PMID- 19562663 TI - Biological control of Lycoriella ingenua (Diptera: Sciaridae) in commercial mushroom (Agaricus bisporus) cultivation: a comparison between Hypoaspis miles and Steinernema feltiae. AB - BACKGROUND: Mushroom cultivation may be adversely affected by insect pests, including sciarids (Lycoriella spp.), which were previously controlled by application of chemical pesticides. However, owing to food safety and environmental concerns, availability of pesticides for use during mushroom cultivation has diminished. Consequently, it is imperative to investigate alternative control strategies, not reliant on chemical pesticides, which may be used in an integrated pest management system. RESULTS: Application of the predatory mite Hypoaspis miles Berlese to commercial mushroom-growing beds at the beginning of spawn run or just prior to casing (830 mites m(-2)) significantly reduced immature sciarids, Lycoriella ingenua (Dufour), in the growing substrate and also adult activity towards the conclusion of cropping. A trend towards lower sciarid emergence from substrates and reduced adult sciarid activity was observed following the application of Steinernema feltiae (Filipjev) (1.5 x 10(6) nematodes m(-2)) at casing. No significant treatment effects on mushroom yield were observed. However, contamination of the mushroom crop by adult sciarids increased in untreated controls. Application of H. miles required a 12-fold increase in labour when compared with application of S. feltiae. CONCLUSION: Contingent upon the development of an effective application system, H. miles has potential for the biological control of sciarids in commercial mushroom production. PMID- 19562664 TI - Quality of nuchal translucency measurements in The Netherlands: a quantitative analysis. AB - OBJECTIVE: The Dutch Centre for Population Research has specified quality demands for nuchal translucency (NT) measurement in The Netherlands. We performed an analysis of the quality of NT measurement in 2005-2006 and its influence on screening performance. METHODS: This was a retrospective study of records of NT measurements (n = 27,738) obtained between January 2005 and December 2006 retrieved from the Dutch National Institute for Public Health and the Environment (RIVM). The performance of each individual operator was analyzed with regard to the quality standards, which involved calculation of operator-specific median NT multiples of the median (MoM) values. For the entire population of operators, a curve was determined describing the relationship between crown-rump length and NT. Detection rates (DR) and false-positive rates (FPR) for Down syndrome were modeled with this new curve and compared to those originally obtained using previously published reference data. RESULTS: Only 22% of all operators met the quality requirement of performing more than 150 NT measurements per year. However, no relationship was found between the number of measurements per operator and their median NT-MoM. The mean of all operator-specific median NT-MoM values was 0.94 (target value 1.0). Overall, operators with The Fetal Medicine Foundation certificate measured a significantly higher median NT-MoM (mean of operator-specific medians, 0.98) as compared to the non-certified operators (0.92). During the study period, the monthly median NT-MoM of all operators rose steadily, from 0.86 in January 2005 to 0.96 in December 2006. Recalculation of the risk for Down syndrome after adjusting the reference NT medians using our own data led to a modeled 4% increase in DR at a 5% FPR. CONCLUSION: Improved monitoring of NT measurement put into effect during the study period seems to have led to an improvement in the accuracy of measurements. Strict quality demands, continued monitoring and scrupulous evaluation of individual operators is likely to lead to an even better performance. PMID- 19562665 TI - Relationship of intertwin crown-rump length discrepancy to chorionicity, fetal demise and birth-weight discordance. AB - OBJECTIVES: To study the frequency and clinical significance of crown-rump length (CRL) discrepancy at 11-14 weeks of gestation in twin pregnancies from an unselected population. METHODS: This was a retrospective analysis of all twin pregnancies that underwent a routine 11-14-week scan at a large teaching hospital. Fetal loss was defined as fetal demise of one or both twins after 14 weeks. RESULTS: A total of 507 twin pregnancies were studied; 382 (75.3%) were dichorionic and 125 (24.7%) were monochorionic twins. The discrepancy in CRL was expressed as a percentage of the CRL of the larger twin. The 95(th) and 99(th) centile for CRL discrepancy in twins was 12.2% and 19.3%, respectively. The discrepancy in CRLs in monochorionic and dichorionic twins was not significantly different (Mann-Whitney U = 22,406, P = 0.302). In 39 twin pairs, there was subsequent intrauterine death of one or both twins. Fetal loss was more common in monochorionic twins (24/125) than in dichorionic twins (15/382) (chi-square = 30.9, P < 0.001). In monochorionic twins, the discrepancy in CRLs in the 24 cases with subsequent loss was significantly greater than in the 101 twin pairs with no subsequent loss (Mann-Whitney U = 896, P = 0.048). The discrepancy in CRLs in 15 dichorionic twins with subsequent loss was not different from that in the 367 twins with no loss (Mann-Whitney U = 2116.5, P = 0.129). The CRL discrepancy was significantly correlated with birth-weight discordance in twins (Spearman's rho = 0.128, P = 0.006). However, this was due to a significant correlation in dichorionic twins (Spearman's rho = 0.127, P = 0.016) but not in monochorionic twins (Spearman's rho = 0.145, P = 0.14). CONCLUSIONS: Fetal loss is significantly associated with discrepancy in CRL at the 11-14-week scan in monochorionic twins and discordance in birth weights is significantly associated with discrepancy in CRL in dichorionic twins. However, intertwin CRL discrepancy is of limited value in screening for these adverse events. PMID- 19562667 TI - PA-MSHA inhibits proliferation and induces apoptosis through the up-regulation and activation of caspases in the human breast cancer cell lines. AB - To investigate the effects of PA-MSHA (Pseudomonas aeruginosa-mannose sensitive hemagglutinin) on inhibiting proliferation of breast cancer cell lines and to explore its mechanisms of action in human breast cancer cells. MCF-10A, MCF-7, MDA-MB-468, and MDA-MB-231HM cells were treated with PA-MSHA or PA (Heat-killed P. aeruginosa) at different concentrations and different times. Changes of cell super-microstructure were observed by transmission electron microscopy. Cell cycle distribution and apoptosis induced by PA-MSHA were measured by flow cytometry (FCM) with PI staining, ANNEXIN V-FITC staining and Hoechst33258 staining under fluorescence microscopy. Western blot was used to evaluate the expression level of apoptosis-related molecules. A time-dependent and concentration-dependent cytotoxic effect of PA-MSHA was observed in MDA-MB-468 and MDA-MB-231HM cells but not in MCF-10A or MCF-7 cells. The advent of PA-MSHA changed cell morphology, that is to say, increases in autophagosomes, and vacuoles in the cytoplasm could also be observed. FCM with PI staining, ANNEXIN V FITC and Hoechst33258 staining showed that the different concentrations of PA MSHA could all induce the apoptosis and G(0)-G(1) cell cycle arrest of breast cancer cells. Cleaved caspase 3, 8, 9, and Fas protein expression levels were strongly associated with an increase in apoptosis of the breast cancer cells. There was a direct relationship with increased concentrations of PA-MSHA but not of PA. Completely different from PA, PA-MSHA may impart antiproliferative effects against breast cancer cells by inducing apoptosis mediated by at least a death receptor-related cell apoptosis signal pathway, and affecting the cell cycle regulation machinery. PMID- 19562668 TI - On the road to understanding of the osteoblast adhesion: cytoskeleton organization is rearranged by distinct signaling pathways. AB - Pre-osteoblast adhesion attracts increasing interest in both medicine and dentistry. However, how this physiological event alters osteoblast phenotype is poorly understood. We therefore attempted to address this question by investigating key biochemical mechanism that governs pre-osteoblast adhesion on polystyrene surface. Importantly, we found that cofilin activity was strongly modulated by PP2A (Ser/Thr phosphatase), while cell-cycle was arrested. Accordingly, we observed that the profile of cofilin phosphorylation (at Ser03) was similar to phospho-PP2A (at Tyr307). Also, it is plausible to suggest during pre-osteoblast adhesion that PP2A phosphorylation at Y307 was executed by phospho Src (Y416). In addition, it was observed that MAPKp38, but not MAPK-erk, played a key role on pre-osteoblast adhesion by phosphorylating MAPKAPK-2 and ATF-2 (also called CRE-BP1). Also, the up-modulation of RhoA reported here suggests its involvement at the beginning of osteoblast attachment, while Akt remained active during all periods. Altogether, our results clearly showed that osteoblast adhesion is under an intricate network of signaling molecules, which are responsible to guide their interaction with substrate mainly via cytoskeleton rearrangement. PMID- 19562669 TI - Matrin 3: chromosomal distribution and protein interactions. AB - Matrin 3 (matr3), an abundant protein of the internal nuclear matrix, has been linked to a variety of functional events. As a step toward defining its multifunctional nature, we have studied the association of matr3 with chromosome territories and identified potential interacting proteins. A similar staining pattern of matr3 was observed in fixed WI38 fibroblast cells and in live HeLa cells using a matr3-GFP construct. Matr3 was detected throughout autosomal and the active X chromosome territories. Conversely, matr3 was strikingly excluded from the inactive X chromosome as well as within both the perinuclear and perinucleolar heterochromatin. Yeast two hybrid analysis identified matr3 interactions with 33 unique nuclear localized proteins and also revealed its propensity for self association. A majority of these proteins are involved in RNA metabolism and chromatin remodeling while others function in protein translation, DNA replication/repair and apoptosis. Further analysis of a selection of these proteins and scaffold attachment factor A (SAFA) by co-localization and co immunoprecipitation experiments using HeLa cells confirmed their interactions with matr3. PMID- 19562670 TI - Celecoxib enhanced the sensitivity of cancer cells to anticancer drugs by inhibition of the expression of P-glycoprotein through a COX-2-independent manner. AB - The P-glycoprotein (p170, P-gp) encoded by human MDR1 gene functions as a pump to extrude anticancer drugs from cancer cells. Over-expression of p170 is closely related to primary and induced drug resistance phenotype of tumor cells. Recent studies have demonstrated that expression of cyclooxygenase-2 (COX-2) is positively correlated with the p170 level, suggesting a potential of COX-2 specific inhibitors in regulation of cytotoxicity of anticancer agents. Celecoxib is one of the specific inhibitors of COX-2 and has been widely used in clinic. However, its function in the response of cancer cells to anticancer drugs and the related mechanism are still waiting to be investigated. To explore the correlation of celecoxib and the p170-mediated drug resistance, the role of celecoxib in drug response of cancer cells was analyzed with flow cytometry, high performance liquid chromatography (HPLC), and colony formation experiments. Celecoxib (50 microM) was found to significantly enhance the sensitivity of MCF-7 and JAR/VP16 cells to tamoxifen and etoposide, respectively, by inhibition of p170 expression and increase in intracellular accumulation of the drugs. However, celecoxib did not affect pump function of p170. Enzyme activity and methylation analyses demonstrated that the inhibitory effect of celecoxib on p170 was independent on COX-2 but closely related to hypermethylation of MDR1 gene promoter. Our study suggested that celecoxib was a potential agent for enhancement of the sensitivity of cancer cells to anticancer drugs. It also provided a links between epigenetic change of MDR1 and drug response of cancer cells. PMID- 19562671 TI - Wnt antagonist SFRP3 inhibits the differentiation of mouse hepatic progenitor cells. AB - Wnt/beta-catenin pathway plays an important role in regulating embryonic development. Hepatocytes differentiate from endoderm during development. Hepatic progenitor cells (HPCs) have been isolated from fetal liver and extrahepatic tissues. Most current studies in liver development and hepatic differentiation have been focused on Wnts, beta-catenin, and their receptors. Here, we sought to determine the role of Wnt antagonists in regulating hepatic differentiation of fetal liver-derived HPCs. Using mouse liver tissues derived from embryonic day E12.5 to postnatal day (PD) 28, we found that 13 of the 19 Wnt genes and almost all of Wnt receptors/co-receptors were expressed in most stages. However, Wnt antagonists SFRP2, SFRP3, and Dkk2 were only detected in the early stages. We established and characterized the reversible stable HPCs derived from E14.5 mouse fetal liver (HP14.5). HP14.5 cells were shown to express high levels of early liver progenitor cell markers, but low levels or none of late liver markers. HP14.5 cells were shown to differentiate into mature hepatocytes upon dexamethasone (Dex) stimulation. Dex-induced late marker expression and albumin promoter activity in HP14.5 cells were inhibited by exogenous expression of SFRP3. Furthermore, Dex-induced glycogen synthesis of PAS-positive HP14.5 cells was significantly inhibited by SFRP3. Therefore, our results have demonstrated that the expression of Wnt antagonists decreases as hepatic differentiation progresses, suggesting that a balanced Wnt signaling may be critical during mouse liver development and hepatic differentiation. PMID- 19562672 TI - A new mechanism of gastric epithelial injury induced by acid exposure: the role of Egr-1 and ERK signaling pathways. AB - The molecular mechanisms by which gastric acid causes epithelial injury in the stomach and initiates an inflammatory reaction are poorly understood. We aimed in the present study to investigate the role of the early growth response gene Egr-1 and ERK in gastric epithelial cells following acid exposure, and the signaling pathways involved. Western blotting was used to assess Egr-1 protein levels in AGS cells. A quantitative measurement of acid-induced Egr-1 and ERK translocation was performed using a high content analysis approach. Egr-1 functionality was assessed by transient transfection with Egr-1 antisense oligonucleotide. Exposure of AGS cells to acidic conditions induced Egr-1 protein expression in a pH- and time-dependent manner. Egr-1 expression was markedly increased as the pH was reduced from pH 7.4 to 6.4. High content analysis of Egr-1 activation showed acid induced Egr-1 nuclear translocation; a maximum observed at 1-2 h followed by a decline to basal levels beyond 4 h. Acidic pH also activated ERK1/2 phosphorylation, whereas ERK1/2 inhibitors PD98059 and U0216 blocked both acid induced Egr-1 and ERK translocation and expression. Moreover, acid exposure up regulated VEGF expression, which was inhibited by the Egr-1 antisense oligonucleotide. Our results also demonstrate that exposure to acid induces Egr-1 via MEK-ERK1/2 pathway. These data suggest that Egr-1 activation might play a crucial role in gastric mucosal inflammation and associated epithelial injury. PMID- 19562673 TI - NF-kappa B is required for the development of tumor spheroids. AB - Tumor cells cultured in three-dimensional models provide a more realistic and biologically meaningful analysis of the initial phases of cancer development and drug resistance. Several studies have demonstrated that culture of cancer cells in three dimensions induces cellular resistance to a variety of anti-neoplastic drugs by poorly understood mechanisms. The role of the transcription factor NF kappaB and inhibitors of apoptosis proteins (IAPs) in the onset and development of drug resistance during tumor spheroid growth has not been established. In this work, we found a significant increase in the activity and expression of NF-kappaB and its downstream target XIAP (X-linked IAP) in cancer cells grown as multi cellular tumor spheroids. Blocking XIAP expression with RNA interference markedly increased the sensitivity of cancer tumor spheroid cells toward anti-neoplastic drugs, indicating a role for IAPs in establishing drug resistance. In turn, inhibition of NF-kappaB by negative dominants suppressed spheroid formation, whereas overexpression of the upstream kinase IkappaBKbeta increased their growth and resistance. The present data suggested that NF-kappaB and its downstream target XIAP were essential for the growth and drug resistance of small avascular tumor. PMID- 19562674 TI - Polyamines affect histamine synthesis during early stages of IL-3-induced bone marrow cell differentiation. AB - Mast cells synthesize and store histamine, a key immunomodulatory mediator. Polyamines are essential for every living cell. Previously, we detected an antagonistic relationship between the metabolisms of these amines in established mast cell and basophilic cell lines. Here, we used the IL-3-driven mouse bone marrow-derived mast cell (BMMC) culture system to further investigate this antagonism in a mast cell model of deeper physiological significance. Polyamines and histamine levels followed opposite profiles along the bone marrow cell cultures leading to BMMCs. alpha-Difluoromethylornithine (DFMO)-induced polyamine depletion resulted in an upregulation of histidine decarboxylase (HDC, the histamine-synthesizing enzyme) expression and activity, accompanied by increased histamine levels, specifically during early stages of these cell cultures, where an active histamine synthesis process occurs. In contrast, DFMO did not induce any effect in either HDC activity or histamine levels of differentiated BMMCs or C57.1 mast cells, that exhibit a nearly inactive histamine synthesis rate. Sequence-specific DNA methylation analysis revealed that the DFMO-induced HDC mRNA upregulation observed in early bone marrow cell cultures is not attributable to a demethylation of the gene promoter caused by the pharmacological polyamine depletion. Taken together, the results support an inverse relationship between histamine and polyamine metabolisms during the bone marrow cell cultures leading to BMMCs and, moreover, suggest that the regulation of the histamine synthesis occurring during the early stages of these cultures depends on the concentrations of polyamines. PMID- 19562675 TI - Thyroid hormones induce cell proliferation and survival in ovarian granulosa cells COV434. AB - Numerous evidences indicate that thyroid hormones exert an important role in the regulation of the reproductive system in the adult female. Although a clear demonstration of the thyroid-ovarian interaction is still lacking, it is conceivable that thyroid hormones might have a direct role in ovarian physiology via receptors in granulosa cells. In this study we analyzed if thyroid hormone treatment could affect cell proliferation and survival of COV434 cells. To this aim cell growth experiments and cell cycle analyses by flow cytometry were performed. Secondly the T(3) survival action was tested by TUNEL assay and MD30 cleavage analysis. We showed that T(3), and not T(4), can protect ovarian granulosa cells COV434 from apoptosis, regulating cell cycle and growth in the same cells. The increase in cell growth resulted in an augmented percentage of the cells in the S phase and, in a reduction of the doubling time (18%). Subsequently apoptotic pathway induced by serum deprivation has been evaluated in the cells exposed or not to thyroid hormone treatment. The T(3) treatment was able to remarkably counteract the apoptotic process. Even at the ultrastructural level there was an evident protective effect of T(3) in the cells that, besides the maintenance of the original morphology and, the absence of basophilic cytoplasm, conserved normal junctional areas. Furthermore, the protective T(3) effect evaluated by FACS analysis in the presence of a PI3K inhibitor revealed, as also confirmed by Western Blot on pAkt, that the PI3K pathway is crucial in T(3) survival action. PMID- 19562677 TI - Hyperglycemia and mechanical stress: targeting the renal podocyte. AB - Hyperglycemia and deriving from glomerular hypertension mechanical stress are the key factors underlying pathogenesis of diabetic nephropathy (DN). Multiple direct and secondary effects of both these factors are mediated by complex signaling pathways with extensive interactions. The common signaling pathways stimulated by high glucose and mechanical insult may act in an additive manner, thereby accelerating the cell damage. Podocytes, the cells covering the outer aspect of glomerular basement membrane (GBM), are subjected not only to the load of filtered glucose but also to diverse mechanical forces. Bulging into the Bowman's space, they have no support from the apical side, which makes them particularly susceptible to the effects of mechanical strain. Both high glucose and mechanical stress may impair the protein systems anchoring the podocyte foot processes in GBM, therefore blunting resistance of these cells to mechanical forces. Modulation by these factors of expression and activity of numerous structural and functional proteins results in the (auto)inflammatory responses, dysfunction, apoptosis or necrosis of the podocytes. Loss of the podocytes is irreversible due to their inability to proliferate and to replenish damaged cells. Podocytes are injured early in the course of DN, which, most likely, underlies further glomerular and renal damage in diabetes. This review summarizes the effects of elevated glucose and mechanical stress that seem to be involved in podocyte impairment in diabetes, with particular focus on the possible interactions between these factors. PMID- 19562678 TI - E2F4 expression is required for cell cycle progression of normal intestinal crypt cells and colorectal cancer cells. AB - The generation of knock-out mice for E2F4 gene expression has suggested a role for this transcription factor in establishing and/or maintaining the intestinal crypt compartment. Having previously demonstrated that E2F4 is cytoplasmic in quiescent-differentiated cells but nuclear in growth factor-stimulated proliferative cells, the present study was aimed at determining the role of E2F4 in the control of human intestinal epithelial proliferation. Results herein demonstrate that lentiviral infection of an shRNA which specifically knocked-down E2F4 expression slowed down G1/S phase transition and the proliferation rate of normal human intestinal epithelial cells (HIEC) and of colon cancer cells. Protein expression of Cdk2, cyclins D1 and A, Cdc25A and c-myc was markedly down regulated in shE2F4-expressing cells; by contrast, expression of the cell cycle inhibitors p21(Cip/Waf) and p27(Kip1) was increased. In addition, the expression of many genes involved in DNA synthesis was down-regulated in shE2F4-expressing cells, whereas no modulation in E2F1 expression was observed. A decrease in E2F4 in colon cancer cell lines also resulted in a reduction in soft-agar growth capacity. Immunofluorescence experiments in human fetal intestine revealed that cells expressing high nuclear levels of E2F4 also expressed cyclin A protein. Lastly, E2F4 and its target cyclin A were up-regulated and mostly nuclear in human colorectal tumor cells in comparison to the corresponding benign epithelium. These results indicate that nuclear E2F4 may be determinant in the promotion of proliferation of human intestinal epithelial crypt cells and colorectal cancer cells. PMID- 19562676 TI - Neural stem cells in the developing and adult brains. AB - Neural stem cells exist in the mammalian developing and adult nervous system. Recently, tremendous interest in the potential of neural stem cells for the treatment of neurodegenerative diseases and brain injuries has substantially promoted research on neural stem cell self-renewal and differentiation. Multiple cell-intrinsic regulators coordinate with the microenvironment through various signaling pathways to regulate neural stem cell maintenance, self-renewal, and fate determination. This review focuses on essential intracellular regulators that control neural stem cell maintenance and self-renewal in both embryonic brains and adult nervous system. These factors include the orphan nuclear receptor TLX, the high-mobility-group DNA binding protein Sox2, the basic helix loop-helix transcription factor Hes, the tumor suppressor gene Pten, the membrane associated protein Numb, and its cytoplasmic homolog Numblike. The aim of this review is to summarize our current understanding of neural stem cell regulation through these important stem cell regulators. PMID- 19562679 TI - Artemisinin--a possible CYP2B6 probe substrate? AB - AIM: To compare in vitro metabolism rates for artemisinin and the CYP2B6 substrates, bupropion, propofol and efavirenz in human liver microsomes. METHODS: Rate constants of artemisinin, bupropion, propofol and efavirenz metabolism by human liver microsomes from a panel of 12 donors, with different levels of CYP2B6 activity, were estimated in WinNonlin. Correlations between the metabolic rate constant for artemisinin and the other CYP2B6 substrates were examined. RESULTS: Artemisinin and propofol depletion data in human liver microsomes were described by first order kinetic models. For bupropion and efavirenz, metabolite formation data were incorporated in the model. Rate constants varied considerably for all substrates. There was a high degree of correlation of rate constants between substrates (r> or =0.87, p<0.001). CONCLUSIONS: The rate of in vitro metabolism of artemisinin was correlated significantly to that of bupropion, propofol and efavirenz, suggesting artemisinin to be a potential alternative marker to assess CYP2B6 activity. Further studies characterizing the metabolic fate of artemisinin are needed in order to evaluate its utility as an in vitro and in vivo CYP2B6 probe substrate, since CYP2B6 might not be the only CYP isoform involved in the depletion of artemisinin. PMID- 19562680 TI - Transport characteristics of candesartan in human intestinal Caco-2 cell line. AB - The intestinal absorptive characteristics and the efflux mechanisms of candesartan (CDS), a novel angiotensin II type 1 receptor blocker, were investigated. The Caco-2 cells were used as models of the intestinal mucosa to assess uptake and transport of CDS. The determination of CDS was performed by HPLC-Flu. In the Caco-2 cells, the uptake and absorptive transport of CDS were pH independent (in the pH range 6.0-8.0). Passive membrane diffusion dominates the absorptive transport behavior of CDS across Caco-2 cells, while secretory transport was a concentration-dependent and saturable process. In the presence of cyclosporin A and verapamil, potent inhibitors of P-glycoprotein (P-gp), the P(ratio) decreased from 3.8 to 2.3 and 1.8, respectively, and permeation of apical to basolateral was enhanced. Overall, the current study suggests that efflux transporters are capable of mediating the absorption and secretion of CDS, and they may play significant roles in limiting the oral absorption of CDS. PMID- 19562681 TI - Expression and regulation of the bile acid transporter, OSTalpha-OSTbeta in rat and human intestine and liver. AB - The regulation of the OSTalpha and OSTbeta expression was studied in the rat jejunum, ileum, colon and liver and in human ileum and liver by ligands for the farnesoid X receptor (FXR), pregnane X receptor (PXR), vitamin D receptor (VDR) and glucocorticoid receptor (GR) using precision cut tissue slices. The gradient of protein and mRNA expression in segments of the intestine for rOSTalpha and rOSTbeta paralleled that of rASBT. OSTalpha and OSTbeta mRNA expression, quantified by qRT-PCR, in rat jejunum, ileum, colon and liver, and in human ileum and liver was positively regulated by FXR and GR ligands. In contrast, the VDR ligand, 1,25(OH)2D3 decreased the expression of rOSTalpha-rOSTbeta in rat intestine, but had no effect on human ileum, and rat and human liver slices. Lithocholic acid (LCA) decreased the expression of rOSTalpha and rOSTbeta in rat ileum but induced OSTalpha-OSTbeta expression in rat liver slices, and human ileum and liver slices. The PXR ligand, pregnenolone-16alpha carbonitrile (PCN) had no effect. This study suggest that, apart from FXR ligands, the OSTalpha and OSTbeta genes are also regulated by VDR and GR ligands and not by PXR ligands. This study show that VDR ligands exerted different effects on OSTalpha-OSTbeta in the rat and human intestine and liver compared with other nuclear receptors, FXR, PXR, and GR, pointing to species- and organ-specific differences in the regulation of OSTalpha-OSTbeta genes. PMID- 19562682 TI - Tissue distribution of the novel DPP-4 inhibitor BI 1356 is dominated by saturable binding to its target in rats. AB - BI 1356 (INN: linagliptin) is an inhibitor of dipeptidyl peptidase-4 (DPP-4). This study investigated whether saturable binding of BI 1356 to its target DPP-4 occurs in tissues and whether drug accumulation occurs at these sites in vivo. In order to test these hypotheses, the tissue distribution of BI 1356 was determined in wild-type and DPP-4 deficient rats at different dose levels by means of whole body autoradiography and measurement of tissue radioactivity concentrations after single i.v. dosing of [(14)C]-radio labeled BI 1356. The accumulation behavior of drug-related radioactivity in tissues was further explored in an oral repeat dose study. Tissue levels of [(14)C]BI 1356 related radioactivity were markedly lower in all investigated tissues of the DPP-4 deficient rats and the difference of the dose-dependent increase of radioactivity tissue levels between both rat strains indicates that tissue distribution at low doses of BI 1356 is dominated by binding of BI 1356 to DPP-4 in tissues. As the binding to DPP-4 is strong but reversible, the tissue binding results in a long terminal half-life in several tissues including plasma. The binding capacity to DPP-4 is, however, limited. In the rat, saturation of DPP-4 binding is suggested at an intravenous dose above 0.01-0.1 mg/kg [(14)C]BI 1356. As the DPP-4 binding capacity is saturated already at low doses, accumulation of BI 1356 in tissues is unlikely, despite the long persistence of low amounts in the body. PMID- 19562683 TI - A critical appraisal of factors affecting the accuracy of results obtained when using flow cytometry in stem cell investigations: where do you put your gates? AB - Flow cytometry is used extensively in stem cell investigations but there is wide variation in the methods used for data analysis between laboratories. Data analysis can be challenging in stem cell biology as there is often no clear distinction between positive and negative populations. We have undertaken a critical appraisal of factors that affect the accuracy of results in stem cell applications. We used mouse embryonic stem cells and determined the expression of three common antigens in stem cell investigations, namely CD15, CD184, and c-kit. We have compared different cell preparation methods and gating strategies and also evaluated the use of isotype controls and unstained cells as controls for the identification of positive populations. The use of a "doublet discriminator" using a side scatter area signal versus side scatter height signal dot plot to identify single cells for analysis increases the accuracy of results regardless of the method used to dissociate cells. Isotype controls can be helpful in mimicking cellular nonspecific binding of the experimental antibody reaction. Isotype controls behave differently on stem cells at different stages of differentiation. Analysis of a viable single cell population with careful selection of control cell populations increases the accuracy of results. PMID- 19562684 TI - Cl-IB-MECA enhances TRAIL-induced apoptosis via the modulation of NF-kappaB signalling pathway in thyroid cancer cells. AB - Apoptosis is an endogenous process that can be a useful anti-cancer tool. This study aimed to investigate the effect of Cl-IB-MECA, adenosine receptor A3 agonist, on TRAIL-induced apoptosis of thyroid carcinoma cells. Cl-IB-MECA enhanced TRAIL-mediated apoptosis in FRO but not in ARO cells. This effect was correlated to higher expression levels of DR5 on FRO than ARO cells, that instead presented higher levels of decoy receptors, DcR1 and DcR2. To understand the cross-talk between the effect of Cl-IB-MECA and TRAIL, we evaluated the nuclear translocation of p65 and c-Rel. Since the dependency by NF-kappaB, TRAIL promoted the nuclear translocation of both p65 and c-Rel subunits. However, the addition of Cl-IB-MECA led to the predominant translocation of c-Rel after TRAIL addition. Furthermore, Bcl-2, cFLIP and pAkt were lower induced than caspase-3 and -9 in FRO cells. To discriminate a specific effect of TRAIL, we used tumour necrosis factor-alpha (TNF-alpha) with Cl-IB-MECA. In this case, no synergism was observed. In addition, the effect of Cl-IB-MECA was not A3 receptor-dependent since its antagonists, MRS1191 and FA385, failed to block Cl-IB-MECA activity on TRAIL-treated FRO cells. In conclusion, Cl-IB-MECA enhanced TRAIL-mediated apoptosis via NF-kappaB/c-Rel activation and DR5-dependent manner. This study may shed light on a potential drug cocktail that may prove useful as anti-cancer in an in vivo animal model. PMID- 19562685 TI - Non-selective cation channel activity is required for lysophosphatidylcholine induced monocyte migration. AB - Lysophosphatidylcholine (LPC) is a major atherogenic lipid which stimulates the recruitment of monocytes to atherosclerotic lesions. The physiological mechanisms underlying LPC-induced monocyte migration are poorly understood. Here we demonstrate that LPC activates non-selective cation channels, which are significantly involved in LPC-induced chemotaxis of monocytes. External LPC elicited the activation of non-selective cation currents in THP-1 monocytes, which occurred in a G protein and phospholipase C-independent manner. LPC activated currents were almost completely inhibited by Gd(3+), La(3+), and TRAM 34. Furthermore, currents were partially reduced by either 2-aminoethoxydiphenyl borate (2-APB) or ruthenium red, while combined application of 2-APB and ruthenium red abolished LPC-activated currents. The 2-APB-sensitive current component was potentiated by flufenamic acid and Ca(2+)-free extracellular solution, while the ruthenium red-sensitive current component was abolished by capsazepine. This pharmacological profile suggests that LPC simultaneously activates TRPC6 and TRPV1 channels in monocytes. Furthermore, in the presence of Gd(3+), La(3+), TRAM-34, 2-APB, ruthenium red or capsazepine, LPC-induced chemotaxis of monocytes was substantially inhibited, indicating that activation of both channel types is required for optimal migration of LPC-stimulated monocytes. Thus, ion channel inhibition may represent a powerful strategy to attenuate the progression of atherosclerosis by reducing monocyte infiltration. PMID- 19562687 TI - Orderly hematopoietic development of induced pluripotent stem cells via Flk-1(+) hemoangiogenic progenitors. AB - Induced pluripotent stem (iPS) cells, reprogrammed somatic cells with embryonic stem (ES) cell-like characteristics, are generated by the introduction of combinations of specific transcription factors. Little is known about the differentiation of iPS cells in vitro. Here we demonstrate that murine iPS cells produce various hematopoietic cell lineages when incubated on a layer of OP9 stromal cells. During this differentiation, iPS cells went through an intermediate stage consisting of progenitor cells that were positive for the early mesodermal marker Flk-1 and for the sequential expression of other genes that are associated with hematopoietic and endothelial development. Flk-1(+) cells differentiated into primitive and definitive hematopoietic cells, as well as into endothelial cells. Furthermore, Flk-1(+) populations contained common bilineage progenitors that could generate both hematopoietic and endothelial lineages from single cells. Our results demonstrate that iPS cell-derived cells, like ES cells, can follow a similar hematopoietic route to that seen in normal embryogenesis. This finding highlights the potential use of iPS cells in clinical areas such as regenerative medicine, disease investigation, and drug screening. PMID- 19562688 TI - Upregulation of peroxisome proliferator-activated receptor-gamma and NADPH oxidases are involved in restenosis after balloon injury. AB - Restenosis is a major complication of percutaneous transluminal coronary angioplasty (PTCA) and is characterized by increased superoxide formation and accumulation of smooth muscle cells (SMCs). The mechanisms through which peroxisome proliferator-activated receptor-gamma (PPAR-gamma) modulates the pathological process are incompletely defined. In this study, balloon injury of porcine coronary arteries in vivo and cell scraping model in vitro were used to elucidate the pathway via this molecule. PPAR-gamma and NADPH oxidase expression significantly increased both in neointimal hyperplasia after balloon injury or in the cultured SMCs after scraping injury. In vitro, PPAR-gamma agonist 15-deoxy Delta(12,14)-prostagladlin J(2) (15d-PGJ2) decreased cell-scraping-induced superoxide generation through suppression of NADPH oxidase activity via down regulation of p22(phox) and gp91(phox). Furthermore, 15d-PGJ2 could suppress scraping-stimulated proliferation of SMCs. These data demonstrate that upregulation of PPAR-gamma and NADPH oxidases are involved in restenosis and activation of PPAR-gamma can inhibit the NADPH oxidase-dependent superoxide generation in SMCs after injury. These findings will provide a new potential drug target for restenosis after balloon injury. PMID- 19562686 TI - Stem cell pluripotency: a cellular trait that depends on transcription factors, chromatin state and a checkpoint deficient cell cycle. AB - Embryonic stem (ES) and induced pluripotent stem (iPS) cells self-renew and are pluripotent. Differentiation of these cells can yield over 200 somatic cell types, making pluripotent cells an obvious source for regenerative medicine. Before the potential of these cells can be maximally harnessed for clinical applications, it will be necessary to understand the processes that maintain pluripotentiality and signal differentiation. Currently, three unique molecular properties distinguish pluripotent stem cells from somatic cells. These include a unique transcriptional hierarchy that sustains the pluripotent state during the process of self-renewal; a poised epigenetic state that maintains chromatin in a form ready for rapid cell fate decisions; and a cell cycle characterized by an extremely short gap 1 (G1) phase and the near absence of normal somatic cell checkpoint controls. Recently, B-MYB (MYBL2) was implicated in the gene regulation of two pluripotency factors and normal cell cycle progression. In this article, the three pluripotency properties and the potential role of B-Myb to regulate these processes will be discussed. PMID- 19562691 TI - EMRS positively impact neurology practice, but implementation challenges remain. PMID- 19562693 TI - Crowdsourcing scientific innovation. PMID- 19562690 TI - Thioredoxin interacting protein (TXNIP) induces inflammation through chromatin modification in retinal capillary endothelial cells under diabetic conditions. AB - Chronic hyperglycemia and activation of receptor for advanced glycation end products (RAGE) are known risk factors for microvascular disease development in diabetic retinopathy. Thioredoxin-interacting protein (TXNIP), an endogenous inhibitor of antioxidant thioredoxin (TRX), plays a causative role in diabetes and its vascular complications. Herein we investigate whether HG and RAGE induce inflammation in rat retinal endothelial cells (EC) under diabetic conditions in culture through TXNIP activation and whether epigenetic mechanisms play a role in inflammatory gene expression. We show that RAGE activation by its ligand S100B or HG treatment of retinal EC induces the expression of TXNIP and inflammatory genes such as Cox2, VEGF-A, and ICAM1. TXNIP silencing by siRNA impedes RAGE and HG effects while stable over-expression of a cDNA for human TXNIP in EC elevates inflammation. p38 MAPK-NF-kappaB signaling pathway and histone H3 lysine (K) nine modifications are involved in TXNIP-induced inflammation. Chromatin immunoprecipitation (ChIP) assays reveal that TXNIP over-expression in EC abolishes H3K9 tri-methylation, a marker for gene inactivation, and increases H3K9 acetylation, an indicator of gene induction, at proximal Cox2 promoter bearing the NF-kappaB-binding site. These findings have important implications toward understanding the molecular mechanisms of ocular inflammation and endothelial dysfunction in diabetic retinopathy. PMID- 19562689 TI - Mutations and polymorphisms of the skeletal muscle alpha-actin gene (ACTA1). AB - The ACTA1 gene encodes skeletal muscle alpha-actin, which is the predominant actin isoform in the sarcomeric thin filaments of adult skeletal muscle, and essential, along with myosin, for muscle contraction. ACTA1 disease-causing mutations were first described in 1999, when a total of 15 mutations were known. In this article we describe 177 different disease-causing ACTA1 mutations, including 85 that have not been described before. ACTA1 mutations result in five overlapping congenital myopathies: nemaline myopathy; intranuclear rod myopathy; actin filament aggregate myopathy; congenital fiber type disproportion; and myopathy with core-like areas. Mixtures of these histopathological phenotypes may be seen in a single biopsy from one patient. Irrespective of the histopathology, the disease is frequently clinically severe, with many patients dying within the first year of life. Most mutations are dominant and most patients have de novo mutations not present in the peripheral blood DNA of either parent. Only 10% of mutations are recessive and they are genetic or functional null mutations. To aid molecular diagnosis and establishing genotype-phenotype correlations, we have developed a locus-specific database for ACTA1 variations (http://waimr.uwa.edu.au). PMID- 19562694 TI - Progress in our understanding of severe drug-induced liver injury. PMID- 19562695 TI - Older donors: mounting risks for the hepatitis C-infected liver transplant recipient? PMID- 19562696 TI - N-type calcium channels mediate a GABA(B) presynaptic modulation in the corticostriatal synapse in turtle's paleostriatum augmentatum. AB - Spikes population evoked by a paired pulse protocol were used to assess the influence of GABA(A) and GABA(B) receptors agonists and antagonists on the synaptic potentials and in the S2/S1 ratio in a paired pulse (PP) protocol in the cortico-paleostriatum augmentatum synapses of the turtle. GABA(A) agonist, muscimol, decreased the amplitude of synaptic responses whereas the facilitation produced with the PP protocol did not change, suggesting a postsynaptic action for GABA(A) receptors. GABA(B) agonist, baclofen, enhanced paired pulse ratio indicating a presynaptic modulation through the GABA(B) receptor. Selective antagonists for N- and P/Q-type Ca(2+)-channels also enhanced paired pulse ratio, suggesting that any of these channel types may be involved in neurotransmitter release. However, the strong paired pulse facilitation produced by baclofen was occluded by blocking the N-type Ca2+ channels with omega-conotoxin GVIA (1 microM), but not by the blockage of P/Q-type Ca2+ channels with omega-agatoxin TK (400 nM). These data suggest that N and P/Q channels participate in the neurotransmitter release, whereas only N-type Ca2+ channels are involved in the presynaptic modulation of GABA(B) in the corticostriatal synapse of the turtle. PMID- 19562697 TI - Regulation of dynamin 2 and G protein-coupled receptor kinase 2 in rat nucleus accumbens during acute and repeated cocaine administration. AB - Exposure to cocaine causes many neuroadaptations including alterations in several neurotransmitter receptors and transporters. This study investigated potential mechanisms of cocaine-induced receptor and transporter regulation by measuring levels of two proteins involved in receptor and transporter trafficking, dynamin 2 and G protein-coupled receptor kinase 2 (GRK2). Male Fischer rats received three daily injections of cocaine, 15 mg/kg, in a binge-pattern (at 1 h intervals) for 1, 3, or 14 days. Brain regions of interest were collected 30 min after the last injection and proteins measured by Western blot. Acute binge pattern cocaine administration produced a significant increase in both dynamin 2- and GRK2-immunoreactivity (227% and 358% of control) in the nucleus accumbens and GKR2 (150% of control) in the caudate putamen. Tolerance to this effect occurred, as levels of both proteins returned to baseline after 3 days of cocaine. In contrast, dynamin 2 and GRK2 were significantly decreased in the nucleus accumbens after chronic cocaine. This pattern of regulation was unique to the nucleus accumbens and not seen in the frontal cortex or substantia nigra. Pretreatment with either the dopamine (DA) D1 receptor antagonist SCH 23390 or D2 receptor antagonist eticlopride prior to acute cocaine blocked the upregulation of dynamin 2 and GRK2 in the nucleus accumbens. However, only eticlopride was effective in attenuating the decrease in these proteins following chronic cocaine exposure. These results demonstrate that two proteins involved in receptor and transporter trafficking are selectively regulated in the nucleus accumbens following acute versus chronic cocaine exposure, and dopamine receptor activation is required for this regulation. PMID- 19562699 TI - Time-dependent exposure of nicotine and smoke modulate ultrasubcellular organelle localization of dopamine D1 and D2 receptors in the rat caudate-putamen. AB - The correlation of the subcellular localization of dopamine D(1) and D(2) receptors (DA D(1) R, DA D(2) R) with nicotine addiction has not been studied. We demonstrated the ultrasubcellular organelle localization of DA D(1) and D(2) Rs in the caudate-putamen (CPu) area of rat brain in vivo exposed to nicotine (3 mg/day; oral) and passive cigarette smoking (500 ml each; 3 times/day) for 1, 4, and 12 weeks, respectively. Our results revealed DA D(1) R localization in the presynaptic and postsynaptic dendrites, endocytic vesicles, and secretory granules, and DA D(2) R localization in the presynaptic dendrites and vesicles. DA D(1) R immunogold particles were highly decreased in the secretory granules of CPu, and increased in the postsynaptic area and vesicles after prolonged nicotine and smoking exposures, suggesting the strong influence of long time smoking and nicotine exposures on DA D(1) R subcellular organelle localization. DA D(2) R immunoreactivity was comparatively less changed than that of the DA D(1) R. Western blot analysis also showed the differential expression of DA D(1) and D(2) R proteins upon nicotine and smoking exposures as compared to the untreated controls. Taken together, the results for the first time suggests the execution of addictive behavior of nicotine through modulation of mesolimbic dopaminergic system targeting subcellular organelle of DA D(1) and D(2) Rs in the CPu of adult rat brain that may lead to novel therapeutic approaches related to nicotine's neuropsychological disorders including drug addiction. PMID- 19562698 TI - In vitro autoradiography and in vivo evaluation in cynomolgus monkey of [18F]FE PE2I, a new dopamine transporter PET radioligand. AB - This study evaluated the in vitro and in vivo characteristics of a new dopamine transporter (DAT) radioligand, [(18)F]fluoroethyl(FE)PE2I, by autoradiography from postmortem human brain and by positron emission tomography (PET) in three cynomolgus monkeys. In the autoradiography experiments, high [18F]FE-PE2I accumulation was observed in caudate and putamen that was selectively abolished by GBR12909 or beta-CIT but not by maprotiline. High doses of citalopram (>5 microM) also inhibited [18F]FE-PE2I binding in the striatum. In vitro Ki of the radioligand was 12 nM at rodent dopamine transporter. [18F]FE-PE2I brain uptake measured by PET was approximately 4-5% of the injected dose, with highest uptake in striatum followed by midbrain and thalamus, lower uptake in neocortex, and lowest in cerebellum. Peak specific binding in striatum was reached approximately 40 min and in midbrain 20-30 min postinjection. The ratio-to-cerebellum was 7-10 in striatum and 1.5-2.3 in midbrain. BP(ND) measured with simplified reference tissue method using the cerebellum as reference region was 4.5 in striatum and 0.6 in midbrain. No displacement was shown after citalopram or maprotiline administration, while GBR12909 decreased the binding in striatum and midbrain to the level of cerebellum. [18F]FE-PE2I showed relatively fast elimination and metabolism with the presence of two metabolite peaks with similar retention time as the labeled metabolites of [11C]PE2I. [18F]FE-PE2I showed in vivo selectivity for the DAT and compared with [11C]PE2I, it showed faster kinetics and earlier peak equilibrium. The potential influence of the two radiometabolites on PET quantification requires further evaluation. PMID- 19562700 TI - Infliximab treatment for steroid-refractory acute graft-versus-host disease after orthotopic liver transplantation: a case report. AB - Acute graft-versus-host disease (GVHD) following orthotopic liver transplantation is a rare but severe disease with a 75% death rate in adults. Various therapeutic strategies have been proposed for steroid-refractory GVHD, but there is still no consensus. Tumor necrosis factor-alpha is a key inflammatory cytokine involved in acute GVHD physiopathology, and infliximab has shown encouraging results for the treatment of acute GVHD following hematopoietic stem cell transplantation. We report the first case of acute GVHD following liver transplantation that was refractory to steroids and anti-lymphocyte globulin but was successfully treated with infliximab. PMID- 19562701 TI - Hepatosplenic gammadelta T-cell lymphoma after liver transplantation: report of the first 2 cases and review of the literature. AB - Hepatosplenic gammadelta T-cell lymphoma is a rare lymphoproliferative disorder originating from natural killer-like Vdelta1-lymphocytes. This subtype has been described after different types of solid organ transplants. In this article, we describe the first 2 cases after liver transplantation. Both patients had thrombocytopenia with (hepato)splenomegaly but without peripheral lymphadenopathies and sinusoidal infiltration of the liver and spleen by monomorphic gammadelta-lymphocytes on pathological examination. The clinical and pathological findings, immunophenotypical profile, prognosis, and treatment are highlighted. In order to make an early diagnosis, physicians who take care of liver transplant recipients should be aware of the characteristic features of this posttransplant lymphoproliferative disorder. Therefore, a diagnostic algorithm is proposed. PMID- 19562702 TI - Quantification of liver perfusion by dynamic magnetic resonance imaging: experimental evaluation and clinical pilot study. AB - Changes in liver microcirculation are considered essential in assessing ischemia reperfusion injury, which in turn has an impact on liver graft function and outcome following liver transplantation (LTx). The aim of this study was to introduce dynamic magnetic resonance imaging (dMRI) as a new technique for overall quantification of hepatic microcirculation and compare it to perfusion measured by laser Doppler flowmetry (LDF; hepatic artery/portal vein) and thermal diffusion (TD). The study included 3 groups, measuring hepatic blood flow and microcirculation with the help of TD, LDF, and dMRI. In group I (9 landrace pigs; 26 +/- 5 kg), the native liver before and after partial portal occlusion was studied; in group II (6 landrace pigs; 25.5 +/- 4.4 kg), the liver 24 hours after LTx was studied; and in group III (14 patients), the liver on days 4 to 7 following LTx was studied. A close correlation was found between dMRI measurements and TD (r = 0.7-0.9, P < 0.01) in 4 defined regions of interest. Portal blood flow and partial occlusion of the portal vein were accurately detected by LDF flowmetry and correlated well with dMRI (r = 0.95, P < 0.01). In the clinical setting, representative TD measurements in segment 4b of the transplanted liver correlated well with dMRI analysis in other segments. Quantification of the portal blood flow and imaging of the whole liver could be performed simultaneously by dMRI. In conclusion, dMRI has been proved to be a sensitive modality for the quantification of liver microcirculation and hepatic blood flow in experimental and clinical LTx. It allows for a synchronous, noninvasive assessment of macrocirculation and microcirculation of the liver and could become a valuable diagnostic tool in advanced liver surgery and transplantation. PMID- 19562703 TI - Unexplained and prolonged perioperative hypotension after orthotopic liver transplantation: undiagnosed systemic mastocytosis. AB - Arterial vasodilation is common in end-stage liver disease, and systemic hypotension often may develop, despite an increase in cardiac output. During the preparation for and the performance of orthotopic liver transplantation, expected and transient hypotension may be caused by induction agents, anesthetic agents, liver mobilization, or venous clamping. A mild decrease of the already low systemic vascular resistance is often observed, and intermittent use of short acting agents for vasopressor support is not uncommon. In this report, we describe a patient with unexpected and prolonged hypotension due to vasodilation during and after orthotopic liver transplantation. The preoperative end-stage liver disease evaluation, intraoperative events, and intensive care unit course were reviewed, and no cause for the vasodilation and prolonged hypotension was evident. The explant pathology report was later available and showed systemic mastocytosis. We hypothesize that the unexpected hypotension and vasodilation were caused by mast cell degranulation and its systemic effects on arterial tone. PMID- 19562704 TI - Early high peak hepatitis C viral load levels independently predict hepatitis C related liver failure post-liver transplantation. AB - The aim of this study was to examine the importance of the serum hepatitis C viral load within the first year post-liver transplant in determining posttransplant survival. A retrospective analysis of 118 consecutive hepatitis C virus-positive liver transplant recipients who received an allograft from January 1997 to September 2005 was undertaken with a median duration of follow-up of 32.4 months. Univariate and multivariate analyses were used to examine the effects of recipient, donor, surgical, and viral factors on posttransplant outcomes. A total of 620 viral load estimations were undertaken in the first 12 months following transplantation. Patient and graft survival rates at 1, 3, and 5 years were 87.8%, 79.9%, and 70.1% and 87.0%, 79.2%, and 68.2%, respectively. According to multivariate analysis, a peak viral load > or = 10(7) IU/mL (P = 0.004; hazard ratio, 8.68; 95% confidence interval, 2.04-37.02) and exposure to antirejection therapy (P = 0.05; hazard ratio, 2.26; 95% confidence interval, 1.01-5.38) were both independent predictors of diminished patient and graft survival and hepatitis C-related allograft failure. The only other independent predictor of hepatitis C virus-related outcome after transplant was azathioprine use, which was associated with improved outcomes (P = 0.04; hazard ratio, 0.25; 95% confidence interval, 0.07-0.91). A peak viral load in the first year after transplant of >10(8), 10(7) to 10(8), and <10(7) IU/mL was associated with a mean survival of 11.8, 70.6, and 89.1 months respectively (P < or = 0.03). The results emphasize the importance of high viral loads in the early posttransplant period as an independent predictor of recipient outcomes. PMID- 19562705 TI - Outcome of liver transplantation for drug-induced acute liver failure in the United States: analysis of the United Network for Organ Sharing database. AB - Acute liver failure (ALF) is an uncommon but potentially lethal drug-related adverse effect that often leads to liver transplantation (LT) or death. A retrospective cohort study was performed with the United Network for Organ Sharing Standard Transplant Analysis and Research files. Recipients who underwent LT for drug-induced acute liver failure (DIALF) from 1987 through 2006 were analyzed. A total of 661 patients transplanted for DIALF were included in the analysis. The 4 leading implicated drug groups were acetaminophen (n = 265; 40%), antituberculosis drugs (n = 50; 8%), antiepileptics (n = 46; 7%), and antibiotics (n = 39; 6%). One-year estimated survival probabilities were 76%, 82%, 52%, 82%, and 79% for acetaminophen, antituberculosis drugs, antiepileptics, antibiotics, and others, respectively. The lower rate of survival among those exposed to antiepileptics was observed mainly in children. Of the 22 patients less than 18 years old who had ALF due to antiepileptics, 73% died within the first year. The difference in overall survival between acetaminophen-related and non acetaminophen-related ALF was not statistically significant. Patients with acetaminophen-related ALF required dialysis prior to LT at a significantly higher rate than all other drug groups (27% versus 3%-10%, P < 0.0001). According to Cox proportional hazards regression analysis, the independent pretransplant predictors of death after LT were being on life support, DIALF due to antiepileptic drugs at age less than 18, and elevated serum creatinine. In conclusion, the leading drug groups causing LT due to DIALF in the United States were acetaminophen, antituberculosis drugs, antiepileptics, and antibiotics. Children who had ALF due to antiepileptics had a substantially higher risk of death after LT in comparison with other drugs. Patients transplanted for acetaminophen-related ALF required dialysis at a significantly higher rate. Being on life support, DIALF due to antiepileptics (at age less than 18), and elevated serum creatinine were independent pretransplant predictors of poor survival after LT for DIALF. PMID- 19562706 TI - Extended right liver grafts obtained by an ex situ split can be used safely for primary and secondary transplantation with acceptable biliary morbidity. AB - Split liver transplantation (SLT) is clearly beneficial for pediatric recipients. However, the increased risk of biliary complications in adult recipients of SLT in comparison with whole liver transplantation (WLT) remains controversial. The objective of this study was to investigate the incidence and clinical outcome of biliary complications in an SLT group using split extended right grafts (ERGs) after ex situ splitting in comparison with WLT in adults. The retrospectively collected data for 80 consecutive liver transplants using ERGs after ex situ splitting between 1998 and 2007 were compared with the data for 80 liver transplants using whole liver grafts in a matched-pair analysis paired by the donor age, recipient age, indications, Model for End-Stage Liver Disease score, and high-urgency status. The cold ischemic time was significantly longer in the SLT group (P = 0.006). As expected, bile leakage from the transected surface occurred only in the SLT group (15%) without any mortality or graft loss. The incidence of all other early or late biliary complications (eg, anastomotic leakage and stenosis) was not different between SLT and WLT. The 1- and 5-year patient and graft survival rates showed no statistical difference between SLT and WLT [83.2% and 82.0% versus 88.5% and 79.8% (P = 0.92) and 70.8% and 67.5% versus 83.6% and 70.0% (P = 0.16), respectively]. In conclusion, ERGs can be used safely without any increased mortality and with acceptable morbidity, and they should also be considered for retransplantation. The significantly longer cold ischemic time in the SLT group indicates the potential for improved results and should thus be considered in the design of allocation policies. PMID- 19562708 TI - Aprotinin and the risk of thrombotic complications after liver transplantation: a retrospective analysis of 1492 patients. AB - Aprotinin is an antifibrinolytic drug that reduces blood loss during orthotopic liver transplantation (OLT). Case reports have suggested that aprotinin may be associated with an increased risk of thromboembolic complications. Recent studies in cardiac surgery also have suggested a higher risk of renal failure and postoperative mortality. Despite these concerns, no large-scale safety assessment has been performed in OLT. In a retrospective observational study involving 1492 liver transplants, we studied the occurrence of postoperative thromboembolic or thrombotic events and mortality in patients who received aprotinin (n = 907) and patients who did not (n = 585). The overall incidence of hepatic artery thrombosis and central venous complications (pulmonary embolism or inferior vena cava thrombosis) was 3.2% and 0.9%, respectively. In propensity score-adjusted analyses (C-index = 0.79), aprotinin was not associated with an increased risk of hepatic artery thrombosis [odds ratio (OR) = 1.00, 95% confidence interval (CI) = 0.50-2.01, P = 0.86]. Although central venous complications were found more frequently in patients receiving aprotinin, the difference was not statistically significant (OR = 2.95, 95% CI = 0.54-16.23, P = 0.32). In addition, no significant differences were found in 1-year mortality (OR = 1.21, 95% CI = 0.86 1.71, P = 0.32). In conclusion, this study did not demonstrate an increased risk of thrombotic complications or mortality when aprotinin is used during OLT. PMID- 19562707 TI - Worse recent efficacy of antiviral therapy in liver transplant recipients with recurrent hepatitis C: impact of donor age and baseline cirrhosis. AB - We hypothesized that antiviral efficacy [sustained virologic response (SVR)] has improved in recent years in the transplant setting. Our aim was to assess whether the efficacy of pegylated interferon (PegIFN)-ribavirin (Rbv) has improved over time. One hundred seven liver transplant patients [74% men, 55.5 years old (range: 37.5-69.5), 86% genotype 1a or 1b] were treated with PegIFN-Rbv for 355 (16-623) days at 20.1 (1.7-132.6) months after transplantation. Tacrolimus was used in 61%. Sixty-seven percent had baseline F3-F4 (cirrhosis: 20.5%). Donor age was 49 (12-78) years. SVR was achieved in 39 (36.5%) patients, with worse results achieved in recent years (2001-2003: n = 27, 46.5%; 2004: n = 23, 43.5%; 2005: n = 21, 35%; 2006 to January 2007: n = 36, 24%; P = 0.043). Variables associated with SVR in the univariate analysis included donor age, baseline viremia and cirrhosis, bilirubin levels, rapid virologic response and early virologic response (EVR), premature discontinuation of PegIFN or Rbv, and accumulated Rbv dose. In the multivariate analysis, the variables in the model were EVR [odds ratio (OR): 0.08, 95% confidence interval (CI): 0.016-0.414, P = 0.002] and donor age (OR: 1.039, 95% CI: 1.008-1.071, P = 0.01). Variables that had changed over time included donor age, baseline viremia, disease severity (cirrhosis, baseline bilirubin, and leukocyte and platelet counts), interval between transplantation and therapy, and use of growth factors. In the multivariate analysis, variables independently changing were donor age (OR: 1.041, 95% CI: 1.013-1.071, P = 0.004), duration from transplantation to antiviral therapy (OR: 1.001, 95% CI: 1.000-1.001, P = 0.013), and baseline leukocyte count (OR: 1.000, 95% CI: 1.000 1.000, P = 0.034). In conclusion, the efficacy of antiviral therapy with PegIFN Rbv has worsened over time, at least in our center. The increase in donor age and greater proportion of patients treated at advanced stages of disease are potential causes. PMID- 19562709 TI - Survival after liver transplantation for hepatocellular carcinoma in the model for end-stage liver disease and pre-model for end-stage liver disease eras and the independent impact of hepatitis C virus. AB - It has been suggested that hepatitis C virus (HCV) patients with hepatocellular carcinoma (HCC) may have worse outcomes after liver transplantation (LT) because of more aggressive tumor biology. In this study, we determined the post-LT survival of HCC patients with and without HCV using United Network for Organ Sharing data from January 1994 to March 2008. Patients with HCC were stratified into HCV (HCC-HCV) and non-HCV (HCC-non-HCV) groups. In the era before the Model for End-Stage Liver Disease (MELD), there were 1237 HCC patients (780, HCV; 373, non-HCV; 84, unknown HCV status), and during the MELD era, there were 4933 HCC patients (3272, HCV; 1348, non-HCV; 313, unknown). In the pre-MELD era, 5-year graft (58.6% versus 53.7%) and patient (61.7% versus 59.3%) survival rates were marginally higher for HCC-non-HCV patients than for HCC-HCV patients. In the MELD era also, 5-year graft (61.2% versus 55.5%) and patient (63.7% versus 58.2%) survival rates were marginally higher for HCC-non-HCV patients than for HCC-HCV patients. In patients without HCC, pre-MELD and MELD era graft/patient survival rates for non-HCV patients were higher than those for HCV patients. The differences in survival rates for HCC patients with and without HCV were lower than those for non-HCC patients stratified by their HCV status. HCV had no additional negative impact on the post-LT survival of patients with HCC, and this was further confirmed by multivariate analysis. In conclusion, the survival of HCC patients has remained unchanged in the past 2 decades. HCV patients have a lower survival rate than non-HCV patients, regardless of their HCC status, but HCV has no additional negative impact on survival in patients with HCC. PMID- 19562710 TI - Amiodarone pretreatment of organ donors exerts anti-oxidative protection but induces excretory dysfunction in liver preservation and reperfusion. AB - The continuous shortage of organs necessitates the use of marginal organs from donors with various diseases, including arrhythmia-associated cardiac failure. One of the most frequently used anti-arrhythmic drugs is amiodarone (AM), which is given in particular in emergency situations. Apart from its anti-arrhythmic actions, AM provides anti-oxidative properties in cardiomyocytes. Thus, we were interested in whether AM donor pretreatment affects the organ quality and function of livers procured for preservation and transplantation. Donor rats were pretreated with AM (5 mg/kg of body weight) 10 minutes before flush-out of the liver with a cold (4 degrees C) histidine-tryptophan-ketoglutarate solution (n = 8). Livers were then stored for 24 hours at 4 degrees C before ex situ reperfusion with a 37 degrees C Krebs-Henseleit solution for 60 minutes in a nonrecirculating system. At the end of reperfusion, tissue samples were taken for histology and Western blot analysis. Animals with vehicle only (0.9% NaCl) served as ischemia/reperfusion controls (n = 8). Additionally, livers of untreated animals (n = 8) not subjected to 24 hours of cold ischemia served as sham controls. AM pretreatment effectively attenuated lipid peroxidation, stress protein expression, and apoptotic cell death. This was indicated by an AM mediated reduction of malondialdehyde, heme oxygenase-1, and caspase-3 activation. However, AM treatment also induced mitochondrial damage and hepatocellular excretory dysfunction, as indicated by a significantly increased glutamate dehydrogenase concentration in the effluate and decreased bile production. In conclusion, AM donor pretreatment exerts anti-oxidative actions in liver preservation and reperfusion. However, these protective AM actions are counteracted by an induction of mitochondrial damage and hepatocellular dysfunction. Accordingly, AM pretreatment of donors for anti-arrhythmic therapy should be performed with caution. PMID- 19562711 TI - Volumes of liver transplant and partial hepatectomy procedures are independently associated with lower postoperative mortality following resection for hepatocellular carcinoma. AB - Partial hepatectomy for hepatocellular carcinoma (HCC) is a high-risk procedure, especially in the presence of portal hypertension. We assessed whether the volume of hospital liver transplant procedures was associated with lower in-hospital mortality independently of the volume of partial hepatectomy procedures. We queried the Nationwide Inpatient Sample (1998-2005) to identify patients who had undergone partial hepatectomy for HCC and used logistic regression to assess the independent effect of volumes of hospital liver transplant and partial hepatectomy procedures on mortality while adjusting for demographic, clinical, and hospital factors. Overall in-hospital mortality was 7.7%. Patients with portal hypertension experienced higher mortality than those who did not (24.5% versus 5.8%, P < 0.0001). Postoperative mortality benefited from a higher volume of hospital liver transplants (>12 per year) and partial hepatectomy procedures (>5 resections per year). Undergoing partial hepatectomy at a center that performed an effective liver transplant volume (eLTV; >12 transplants per year) was associated with lower mortality in both the portal hypertensive group (16.4% versus 33.7%, P = 0.004) and non-portal hypertensive group (4% versus 8%, P = 0.0002). After multivariate adjustment, the odds ratio (OR) of in-hospital death for those with portal hypertension was 4.5 [95% confidence interval (CI), 2.98 6.81]. The lower mortality observed with eLTV (OR, 0.59; 95% CI, 0.37-0.93) was independent of the mortality benefit from an effective partial hepatectomy volume (>5 hepatectomies per year; OR, 0.54; 95% CI, 0.31-0.94). Postoperative complications were also fewer at centers with eLTV compared to those without eLTV (39.2% versus 29.3%, P < 0.0001). In conclusion, given the postoperative mortality benefit independent of the volume of partial hepatectomy procedures, referral to a center with eLTV should be considered for HCC resection, especially in the presence of portal hypertension. PMID- 19562712 TI - Effects of EGFR tyrosine kinase inhibitor erlotinib in prostate cancer cells in vitro. AB - BACKGROUND: Erlotinib is a small-molecule tyrosine kinase inhibitor targeted EGFR, known to be overexpressed in a variety of cancers, including prostate cancer. Clinical trials showed insignificant clinical benefit in patients with castration resistant prostate cancer both when EGFR inhibitors were administered as monotherapy or in association with antiandrogens or chemotherapeutics. Why, differently to other tumors, have EGFR inhibitors been so ineffective in human prostate cancer? This is the question that we have set in this report. METHODS: For this purpose, the effectiveness of erlotinib, a selective EGFR inhibitor, in a wide range of prostate cancer cells (wild type or engineered to overexpress peculiar proteins including androgen receptor and PTEN). RESULTS: We demonstrated that the effectiveness of erlotinib was inversely correlated to the EGFR/Her2 ratio rather than EGFR/p-EGFR or Her2/p-Her2 levels. Chronic treatment with bicalutamide induced overexpression of Her2 and reduction of EGFR/Her2ratio and this was associated with increased Akt and Erk activity. In these conditions of treatment a reduced efficacy of erlotinib was observed. At the same time, an increased efficacy versus erlotinib was documented in cancer cells chronically exposed to DHT. In these culture conditions low levels of Her2 and increased EGFR/Her2 ratio were evidenced. CONCLUSIONS: Taken together, our results seem to suggest that a low EGFR/Her2 ratio and PTEN absence are the main factors responsible of erlotinib inefficacy. Therefore the inhibition of EGFR could have important antitumor effects in hormone-naive rather than in hormonally treated patients. PMID- 19562713 TI - Chemokine-derived peptides as carriers for gene delivery to CXCR4 expressing cells. AB - BACKGROUND: Cell and tissue-specific DNA delivery can be achieved by derivatizing vehicles with a targeting ligand for certain receptor. CXCR4 is a receptor of chemokine stromal cell-derived factor (SDF)-1 and viral protein viral macrophage inflammatory protein (vMIP)-II. It is expressed on some types of stem and cancer cells. The present study aimed to design and characterize the group of CXCR4 targeted peptides for receptor-mediated gene delivery. We focused on bifunctional peptide carriers: two derived from N-terminal sequences of SDF-1 and one from vMIP-II. METHODS: Three synthetic chemokine-derived peptides, designated long CDP (KPVSLSYRSPSRFFESH-K9-biotin), short CDP (KPVSLSYR-K9-biotin) and viral CDP (D LGASWHRPDK-K9-biotin), were evaluated for gene delivery to CXCR4 positive and negative cells. Oligolysine K9-biotin was used as a control. The Lys 8 moiety binds DNA electrostatically, whereas C-terminal lysine was modified with biotin to study intracellular uptake of the peptides. Complex formation with DNA was monitored by ethidium bromide fluorescence quenching. RESULTS: All peptides condensed plasmid DNA. Gene delivery by CDP/DNA complexes is glycerol-dependent and the level of luciferase expression with signal modified carriers was comparable with the efficacy of polyethylenimine (PEI) in CXCR4 expressing cell lines (A172, HeLa) and was ten- to 50-fold higher compared to unmodified peptide. By contrast, CDP transfection efficacy on CXCR4-negative cells (chinese hamster ovary) was much lower than in PEI. Intracellular uptake analysis of biotin labeled peptides indicated that CDPs entered cells more efficiently than oligolysine. CONCLUSIONS: The small, bifunctional peptides reported in the present study may be useful in gene delivery to (and gene therapy of) the different tumors and other cells expressing CXCR4. PMID- 19562715 TI - Predictors of sustained virological response after antiviral treatment for hepatitis C recurrence following liver transplantation. AB - Factors associated with sustained virological response (SVR) in patients treated for hepatitis C virus (HCV) recurrence after liver transplantation (LT) are unclear. Ninety-nine HCV-positive/hepatitis B surface antigen-negative patients received antiviral treatment (AVT) with interferon/peginterferon plus ribavirin for HCV recurrence after LT. Cyclosporine (CyA) or tacrolimus (TAC) was used as the main immunosuppressor in 37 (37%) and 62 (63%) patients, respectively. Twenty five patients (25%) achieved an SVR. Twenty-seven donor-related, recipient related, HCV-related, and immunosuppression-related variables were investigated for their association with SVR. In logistic regression analysis, donor age < 60 years (odds ratio = 4.45, 95% confidence interval = 1.39-14.19, P = 0.01), viral genotype other than 1 (odds ratio = 4.97, 95% confidence interval = 1.59-15.48, P = 0.006), and the use of CyA during treatment (odds ratio = 6.85, 95% confidence interval = 2.15-21.73, P = 0.001) were predictors of SVR. Patients treated with CyA (SVR rate: 43%) and those treated with TAC (SVR rate: 14%) were comparable for all variables, except for a shorter ischemia time and shorter timing of AVT initiation in the TAC group (P = 0.02 and P = 0.005, respectively) and a greater use of anti-CD25 antibodies, azathioprine, and mycophenolate mofetil in the CyA group (P = 0.03, P < 0.001, and P = 0.001, respectively). The rate of AVT discontinuation due to side effects was similar between groups (16% versus 8%, P = 0.3). In conclusion, the type of immunosuppression during AVT may predict SVR in patients treated for HCV recurrence after LT. PMID- 19562714 TI - Variation in novel exons (RACEfrags) of the MECP2 gene in Rett syndrome patients and controls. AB - The study of transcription using genomic tiling arrays has lead to the identification of numerous additional exons. One example is the MECP2 gene on the X chromosome; using 5'RACE and RT-PCR in human tissues and cell lines, we have found more than 70 novel exons (RACEfrags) connecting to at least one annotated exon.. We sequenced all MECP2-connected exons and flanking sequences in 3 groups: 46 patients with the Rett syndrome and without mutations in the currently annotated exons of the MECP2 and CDKL5 genes; 32 patients with the Rett syndrome and identified mutations in the MECP2 gene; 100 control individuals from the same geoethnic group. Approximately 13 kb were sequenced per sample, (2.4 Mb of DNA resequencing). A total of 75 individuals had novel rare variants (mostly private variants) but no statistically significant difference was found among the 3 groups. These results suggest that variants in the newly discovered exons may not contribute to Rett syndrome. Interestingly however, there are about twice more variants in the novel exons than in the flanking sequences (44 vs. 21 for approximately 1.3 Mb sequenced for each class of sequences, p=0.0025). Thus the evolutionary forces that shape these novel exons may be different than those of neighboring sequences. PMID- 19562716 TI - A randomized pilot trial of oral branched-chain amino acids in early cirrhosis: validation using prognostic markers for pre-liver transplant status. AB - Because of the chronic shortage of liver donors, hepatologists are required to prolong the liver transplant waiting period by preserving the hepatic reserve of scheduled recipients. This study examined the effectiveness of oral branched chain amino acids (BCAAs), using outcome markers indicating pretransplant hepatic reserve. Fifty-six consecutive eligible patients with Child class A cirrhosis without major complications were randomly assigned to receive oral BCAA granules (12.45 g/day) for least 1 year or no BCAAs. Differences between groups in the Model for End-Stage Liver Disease (MELD) score, Child-Turcotte-Pugh (CTP) score, asialoscintigraphic clearance index (CI), and complications were examined. Of 50 remaining patients, 27 received BCAAs, and 23 received no BCAAs (mean duration, 3.2 years). The mean annual changes in the MELD score, CTP score, and asialoscintigraphic CI were smaller in the BCAA group than in the control group ( 0.06 +/- 0.23 versus 0.10 +/- 0.40, P = 0.024, 0.06 +/- 0.30 versus 0.30 +/- 0.48, P = 0.037, and 0.00 +/- 0.02 versus 0.02 +/- 0.04, P = 0.040, respectively). The mean annual changes in the serum total bilirubin and the serum albumin in the BCAA group were better preserved than those in the control group ( 0.07 +/- 0.20 versus 0.12 +/- 0.18 mg/dL, P < 0.001, and 0.07 +/- 0.13 versus 0.02 +/- 0.19 g/dL, P = 0.005, respectively); other laboratory variables were not significant. The incidence of overall major cirrhotic complications was lower in the BCAA group than in the control group [14.8% (4 of 27 patients) versus 30.4% (7 of 23 patients) at 3 years, P = 0.043]; only ascites was significant individually. In conclusion, early interventional oral BCAAs might prolong the liver transplant waiting period by preserving hepatic reserve in cirrhosis. PMID- 19562717 TI - Impaired autophagic clearance after cold preservation of fatty livers correlates with tissue necrosis upon reperfusion and is reversed by hypothermic reconditioning. AB - Fatty livers are particularly susceptible to mitochondrial alterations after cold preservation. We thus aimed to improve graft integrity by brief hypothermic oxygenation prior to warm reperfusion. Macrovesicular steatosis was induced in rat livers by fasting and subsequent feeding of a fat-free diet enriched with carbohydrates. Fatty livers were retrieved and stored ischemically at 4 degrees C for 20 hours in histidine-tryptophan-ketoglutarate solution. Hypothermic reconditioning (HR) was performed in some livers by insufflation of gaseous oxygen via the caval vein during the last 90 minutes of preservation. Viability was assessed upon isolated reperfusion. HR resulted in a significant (approximately 5-fold) reduction of parenchymal (alanine aminotransferase and lactate dehydrogenase) and mitochondrial (glutamate dehydrogenase) enzyme release. Functional recovery (bile production, oxygen consumption, and tissue levels of adenosine triphosphate) was significantly improved by HR. In untreated grafts, cellular autophagy (cleavage of LC3B and protein expression of beclin-1) was significantly impaired (<50% of baseline) after preservation/reperfusion but was restored to normal values by HR. HR also increased cleavage of caspase 9 (P < 0.5) and caspase 3 enzyme activity (by a factor of 1.5). In contrast, histological signs of tissue necrosis were abundant after reperfusion in untreated livers and largely abrogated in reconditioned livers. In conclusion, HR limits mitochondrial defects and restores basal rates of cellular autophagy. This may represent a rescue mechanism for maintaining cellular homeostasis and tissue survival. PMID- 19562718 TI - Recurrent familial hypobetalipoproteinemia-induced nonalcoholic fatty liver disease after living donor liver transplantation. AB - Familial hypobetalipoproteinemia (FHBL) is one of the causes of nonalcoholic steatohepatitis (NASH) and a codominant disorder. Patients heterozygous for FHBL may be asymptomatic, although they demonstrate low plasma levels of low-density lipoprotein (LDL) cholesterol and apolipoprotein B. Here we report a nonobese 54 year-old man with decompensated liver cirrhosis who underwent living donor liver transplantation with his son as the donor. Low albuminemia and refractory ascites persisted after transplantation. A biopsy specimen obtained 11 months after liver transplantation revealed severe steatosis and fibrosis, and recurrent NASH was diagnosed on the basis of pathological findings. Both the patient's and donor's laboratory tests demonstrated low LDL cholesterol and apolipoprotein levels. Because mutations in messenger RNAs of microsomal triglyceride transfer protein and apolipoprotein B genes were excluded neither in the recipient nor in the donor, both were clinically diagnosed as being heterozygous for FHBL. We successfully treated the recipient with heterozygous FHBL-induced recurrent NASH after liver transplantation using our diet and exercise programs. PMID- 19562719 TI - Transthoracic open window hepatostomy: a salvage approach to right lobe abscesses after liver transplantation. PMID- 19562720 TI - Use of transjugular intrahepatic portosystemic shunts after liver transplantation. PMID- 19562721 TI - Common hepatic duct with five tributaries: a dangerous anomaly in hepatic resection and living donor liver transplantation. PMID- 19562722 TI - Are the Hangzhou criteria adaptable to hepatocellular carcinoma patients for liver transplantation in Western countries? PMID- 19562724 TI - The FOXF2 pathway in the human prostate stroma. AB - BACKGROUND: Forkhead box 2 (FOXF2) is a member of the large family of forkhead transcription factors and its expression pattern suggests a role in prostate cancer development. FOXF2 expression is stroma-specific and higher expressed in the prostate transition zone than the prostate peripheral zone. Moreover, expression of FOXF2 is decreased in prostate cancer. METHODS: To identify the genes and pathways regulated by FOXF2, we compared microarray expression profiles of primary prostate stromal cells (PrSC) treated with control or small interfering RNA (siRNA) directed against FOXF2. RESULTS: From our microarray analyses, we selected 190 differentially expressed genes, of which 104 genes were higher expressed in PrSC cells treated with FOXF2 siRNA and 86 were higher expressed in PrSC cells treated with negative control siRNA. Eight of the strongest differentially expressed genes were validated by RT-PCR. Genes down regulated by FOXF2 included MT1E, MT1F, PDGFA, ITGB1, and PSG7 and genes up regulated by FOXF2 included WASF2, BAMBI, and CXCL12. Ingenuity pathway analysis showed several pathways significantly regulated by FOXF2, including PPAR signaling, PDGF signaling, and extracellular matrix (ECM) signaling. GSEA analysis revealed that FOXF2 up-regulated genes were down-regulated in the same PrSC cells treated with transforming growth factor 3 (TGFbeta3). CONCLUSIONS: The distinct expression pattern of FOXF2 in the prostate, its effect on expression of ECM signaling, and its opposing role in the TGFbeta3 pathway, suggests a role for FOXF2 in prostate homeostasis and stroma-epithelial interactions. PMID- 19562725 TI - Using a partnership between academic faculty and a physical therapist liaison to develop a framework for an evidence-based journal club: a discussion. AB - Evidence-based practice (EBP) in rehabilitation is increasingly recognized as important. Despite the importance of EBP, physical therapists' knowledge of EBP varies. Journal clubs have been used to educate clinicians about EBP. This discussion paper describes the partnership between academic faculty members and a physical therapist at a community hospital, and the process used to develop a framework to implement an evidence-based journal club. The partnership blended the expertise of academic faculty members and a physical therapist with knowledge of EBP who served as the liaison between members of the partnership team and the clinicians at the community hospital. The three-step framework developed enabled the clinicians to learn about critical appraisal, participate in guided practice of critical appraisal with the liaison, and lead critical appraisal of a paper with the assistance of the liaison as needed. This process could be easily replicated by other partnerships between academic faculty members and clinicians. Developing partnerships like the one described enables academicians to provide service to the profession, may enhance physical therapists' knowledge of the principles of EBP and may encourage EBP. PMID- 19562727 TI - Transfer of occupational health problems from a developed to a developing country: lessons from the Japan-South Korea experience. AB - Many corporations move their manufacturing facilities or technologies from developed to developing countries. Stringent regulations have made it costly for industries to operate in developed, industrialized countries. In addition, labor costs are high in these countries, and there is increasing awareness among the general public of the health risks associated with industry. The relocation of hazardous industries to developing countries is driven by economic considerations: high unemployment, a cheaper labor force, lack of regulation, and poor enforcement of any existing regulations make certain countries attractive to business. The transfer of certain industries from Japan to Korea has also brought both documented occupational diseases and a new occupational disease caused by chemicals without established toxicities. Typical examples of documented occupational diseases are carbon disulfide poisoning in the rayon manufacturing industry, bladder cancer in the benzidine industry, and mesothelioma in the asbestos industry. A new occupational disease due to a chemical without established toxicities is 2-bromopropane poisoning. These examples suggest that counter-measures are needed to prevent the transfer of occupational health problems from a developed to a developing country. Corporate social responsibility should be emphasized, close inter-governmental collaboration is necessary and cooperation among non-governmental organizations is helpful. PMID- 19562726 TI - Inhibition of Abeta42 aggregation using peptides selected from combinatorial libraries. AB - Increasing evidence suggests that the aggregation of the small peptide Abeta42 plays an important role in the development of Alzheimer's disease. Inhibiting the initial aggregation of Abeta42 may be an effective treatment for preventing, or slowing, the onset of the disease. Using an in vivo screen based on the enzyme EGFP, we have searched through two combinatorially diverse peptide libraries to identify peptides capable of inhibiting Abeta42 aggregation. From this initial screen, three candidate peptides were selected and characterized. ThT studies indicated that the selected peptides were capable of inhibiting amyloid aggregation. Additional ThT studies showed that one of the selected peptides was capable of disaggregating preformed Abeta42 fibers. PMID- 19562728 TI - Non-invasive determination of tissue thermal parameters from high intensity focused ultrasound treatment monitored by volumetric MRI thermometry. AB - A method is proposed for estimating the perfusion rate, thermal diffusivity, and the absorption coefficient that influence the local temperature during high intensity focused ultrasound (HIFU) thermotherapy procedures. For this purpose, HIFU heating experiments (N = 100) were performed ex vivo on perfused porcine kidney (N = 5) under different flow conditions. The resulting spatio-temporal temperature variations were measured non-invasively by rapid volumetric MR temperature imaging. The bio-heat transfer (BHT) model was adapted to describe the spatio-temporal evolution of tissue temperature in the cortex. Absorption and perfusion coefficients were determined by fitting the integrated thermal load (spatial integration of the thermal maps) curves in time with an analytical solution of the BHT equation proposed for single point HIFU heating. Thermal diffusivity was determined independently by analyzing the spatial spread of the temperature in time during the cooling period. Absorption coefficient and thermal diffusivity were found to be independent of flow, with mean and average values of 11.0 +/- 1.85 mm(3) x K x J(-1) and 0.172 +/- 0.003 mm(2) x s(-1), respectively. A linear dependence of the calculated perfusion rate with flow was observed with a slope of 9.20 +/- 0.75 mm(-3). The perfusion was found to act as a scaling term with respect to temperature but with no effect on the spatial spread of temperature which only depends on the thermal diffusivity. All results were in excellent agreement with the BHT model, indicating that this model is suitable to predict the evolution of temperature in perfused organs. This quantitative approach allows for determination of tissue thermal parameters with excellent precision (within 10%) and may thus help in quantifying the influence of perfusion during MR guided high intensity focused ultrasound (MRgHIFU). PMID- 19562730 TI - Mortality among sheet metal workers participating in a medical screening program. AB - BACKGROUND: The Sheet Metal Occupational Health Institute Trust (SMOHIT) was formed in 1985 to examine the health hazards of the sheet metal industry in the U.S. and Canada through an asbestos disease screening program. A study of mortality patterns among screening program participants was undertaken. METHODS: A cohort of 17,345 individuals with 20 or more years in the trade and who participated in the asbestos disease screening program were followed for vital status and causes of death between 1986 and 2004. Data from the screening program included chest X-ray results by International Labour Office (ILO) criteria and smoking history. Standardized mortality ratios (SMRs) by cause were generated using U.S. death rates and Cox proportional hazards models were used to investigate lung cancer risk relative to chest X-ray changes while controlling for smoking. RESULTS: A significantly reduced SMR of 0.83 (95% CI = 0.80-0.85) was observed for all causes combined. Statistically significant excess mortality was observed for pleural cancers, mesothelioma, and asbestosis in the SMR analyses. Both lung cancer and COPD SMRs increased consistently and strongly with increasing ILO profusion score. In Cox models, which controlled for smoking, increased lung cancer risk was observed among workers with ILO scores of 0/1 (RR = 1.17, 95% CI = 0.89-1.54), with a strong trend for increasing lung cancer risk with increasing ILO profusion score >0/0. CONCLUSIONS: Sheet metal workers are at increased risk for asbestos-related diseases. This study contributes to the literature demonstrating asbestos-related diseases among workers with largely indirect exposures and supports an increased lung cancer risk among workers with low ILO profusion scores. PMID- 19562729 TI - Common variants in 8q24 are associated with risk for prostate cancer and tumor aggressiveness in men of European ancestry. AB - BACKGROUND: Recent whole genome association studies have independently identified multiple prostate cancer (PC) risk variants on 8q24. We have evaluated association of common variants in this region with PC susceptibility and tumor aggressiveness in a sample of European American men. METHODS: Forty-nine tagging SNPs including three previously reported significant variants (rs1447295, rs6983267, rs16901979) and seven variants in the 5' upstream region of the MYC proto-oncogene were tested for association with susceptibility to PC and tumor aggressiveness in 596 histologically verified PC cases and 567 ethnically matched controls. RESULTS: Significant associations with susceptibility to PC were found at 17 SNPs, four of which (rs1016342, rs1378897, rs871135, and rs6470517) remained significant after adjusting for multiple corrections. One of the associated SNPs, rs871135, is located in the putative gene POU5F1P1 within the 8q24 region. An in slico analysis showed that the associated variant of this SNP alters a transcription factor implicating a plausible regulatory role. Additionally, one of the significantly associated SNPs, rs6470517, with PC susceptibility showed a significant over-representation of the G allele in cases with aggressive tumor. CONCLUSIONS: Although this study does not directly confirm associations of the three specific SNPs (cited above), it corroborates reported signals of association in 8q24 reaffirming that genetic variation on 8q24 influences susceptibility to PC in men of European ancestry. Although our study did not confirm the allelic association of rs1447295, meta-analysis of this SNP provided support to previous reported associations. Further, this study implicates the 8q24 region with aggressive forms of PC. PMID- 19562731 TI - Synthesis and assembly of a full-length human monoclonal antibody in algal chloroplasts. AB - Monoclonal antibodies can be effective therapeutics against a variety of human diseases, but currently marketed antibody-based drugs are very expensive compared to other therapeutic options. Here, we show that the eukaryotic green algae Chlamydomonas reinhardtii is capable of synthesizing and assembling a full-length IgG1 human monoclonal antibody (mAb) in transgenic chloroplasts. This antibody, 83K7C, is derived from a human IgG1 directed against anthrax protective antigen 83 (PA83), and has been shown to block the effects of anthrax toxin in animal models. Here we show that 83K7C heavy and light chain proteins expressed in the chloroplast accumulate as soluble proteins that assemble into complexes containing two heavy and two light chain proteins. The algal-expressed 83K7C binds PA83 in vitro with similar affinity to the mammalian-expressed 83K7C antibody. In addition, a second human IgG1 and a mouse IgG1 were also expressed and shown to properly assemble in algal chloroplast. These results show that chloroplasts have the ability to fold and assemble full-length human mAbs, and suggest the potential of algae as a platform for the cost effective production of complex human therapeutic proteins. PMID- 19562732 TI - Dual-label centromere and telomere FISH identifies human, rat, and mouse cell contribution to Multispecies recombinant urogenital sinus xenografts. AB - BACKGROUND: Recombinant xenografts of human cells growing in immunocompromised rodents are widely used for studying stem cell biology, tumor biology, and epithelial to mesenchyme transitions. Of critical importance is the correct interpretation of the cellular composition of such xenografts. METHODS: Here we present a rapid and robust method employing protein nucleic acid (PNA) FISH probes to dual-label centromeres and telomeres (Cen/Tel FISH). Such labeling allows unambiguous discrimination between human, mouse, and rat cells in paraffin embedded tissue sections, providing significant advantages over current methods used to discern human versus rodent cell types. RESULTS: Using an in vivo prostatic developmental system where rat embryonic urogenital sinus mesenchyme is recombined with human prostate epithelial organoids and grown in an immunocompromised mouse, Cen/Tel FISH documents that all three species contribute to the development of glandular structures. CONCLUSIONS: The method is an indispensable tool to analyze xenograft/host interactions and prevent misinterpretation of data using tissue recombination approaches. PMID- 19562733 TI - A multipurpose capacitive biosensor for assay and quality control of human immunoglobulin G. AB - We report a flow-injection biosensor system with a capacitive transducer for assay and quality control of human immunoglobulin G (hIgG). The sensing platform is based on self-assembled monolayers (SAMs) of carboxylic acid terminated alkyl thiols with covalently attached concanavalin A. The electrochemical characteristics of the sensor surface were assessed by cyclic voltammetry using a permeable redox couple (potassium ferricyanide). The developed biosensor proved capable of performing a sensitive label-free assay of hIgG with a detection limit of 1.0 microg mL(-1). The capacitance response depended linearly on hIgG concentration over the range from 5.0 to 100 microg mL(-1), in a logarithmic plot. Typical measurements were performed in 15 min and up to 18 successive assays were achieved without significant loss of sensitivity using a single electrode. In addition, the biosensor can detect hIgG aggregates with concentrations as low as 0.01% of the total hIgG content (5.0 microg mL(-1)). Hence, it represents a potential post-size-exclusion chromatography-UV (post-SEC UV) binding assay for in-process quality control of hIgG, which cannot be detected by SEC-UV singly at concentrations below 0.3% of the total hIgG content. PMID- 19562734 TI - Modeling the effects of obesity and weight gain on PSA velocity. AB - BACKGROUND: Hemodilution theory states that higher blood volume in obese men effectively dilutes circulating PSA levels resulting in lower PSA test results. Here we apply hemodilution theory to model the effect of weight gain on PSA velocity. METHODS: Hemodilution formulas were used to model PSA velocity for a series of plausible scenarios in which initial weight and weight gain were varied. The formulas were also applied to published summary data on weight, weight change and PSA velocity from the Prostate Cancer Prevention Trial (PCPT). RESULTS: Under hemodilution theory, PSA velocity is understood to be influenced by total circulating PSA mass (ng of PSA) and BMI at the initial test and total circulating PSA mass and BMI at the subsequent test. PSA velocity in a man with a stable BMI of 35 is estimated to be 13% lower than in a man with a stable BMI of 25. A gain of 4 BMI units is predicted to attenuate PSA velocity by as much as 25%. When applied to summary data from the PCPT, the formulas estimate that a 10 pound weight gain causes a -0.028 ng/ml change in PSA, which closely matches PCPT results where a 10 pound weight gain caused a -0.024 ng/ml change in PSA. We provide software to implement the hemodilution formulas to model PSA velocity for different weights and changes in weight through time (http://www.eheintl.com/psa.jsp). CONCLUSION: Stable obesity and weight gain both independently attenuate PSA velocity, potentially obscuring clinically relevant changes in circulating PSA. PMID- 19562735 TI - Genome-wide transcriptomic profiling of microdissected human breast tissue reveals differential expression of KIT (c-Kit, CD117) and oestrogen receptor alpha (ERalpha) in response to therapeutic radiation. AB - The pathogenesis of late normal tissue fibrosis after high-dose ionizing radiation involves multiple cell types and signalling pathways but is not well understood. To identify the molecular changes occurring after radiotherapy, paired normal tissue samples were collected from the non-irradiated breast and from the treated breast of women who had undergone curative radiotherapy for early breast cancer months or years previously. As radiation may induce distinct transcriptional changes in the different components of the breast, laser capture microdissection and gene expression microarray profiling were performed separately for epithelial and stromal components and selected genes were validated using immunohistochemistry. In the epithelial compartment, a reduction of KIT (c-Kit; CD117) and a reciprocal increase in ESR1 (oestrogen receptor alpha, ERalpha) mRNA and protein levels were seen in irradiated compared to non irradiated samples. In the stromal compartment, extracellular matrix genes including FN1 (fibronectin 1) and CTGF (connective tissue growth factor; CCN2) were increased. Further investigation revealed that c-Kit and ERalpha were expressed in distinct subpopulations of luminal epithelial cells. Interlobular c Kit-positive mast cells were also increased in irradiated cases not showing features of post-radiation atrophy. Pathway analysis revealed 'cancer, reproductive system disease and tumour morphology' as the most significantly enriched network in the epithelial compartment, whereas in the stromal component, a significant enrichment for 'connective tissue disorders, dermatological diseases and conditions, genetic disorder' and 'cancer, tumour morphology, infection mechanism' networks was observed. These data identify previously unreported changes in the epithelial compartment and show altered expression of genes implicated in late normal tissue injury in the stromal compartment of normal breast tissue. The findings are relevant to both fibrosis and atrophy occurring after radiotherapy for early breast cancer. PMID- 19562736 TI - Estimation of absolute risk for prostate cancer using genetic markers and family history. AB - BACKGROUND: Multiple DNA sequence variants in the form of single-nucleotide polymorphisms (SNPs) have been found to be reproducibly associated with prostate cancer (PCa) risk. METHODS: Absolute risk for PCa among men with various numbers of inherited risk alleles and family history of PCa was estimated in a population based case-control study in Sweden (2,893 cases and 1,781 controls), and a nested case-control study from the Prostate, Lung, Colon and Ovarian (PLCO) Cancer Screening Trial in the U.S. (1,172 cases and 1,157 controls). RESULTS: Increased number of risk alleles and positive family history were independently associated with PCa risk. Considering men with 11 risk alleles (mode) and negative family history as having baseline risk, men who had >or=14 risk alleles and positive family history had an odds ratio (OR) of 4.92 [95% confidence interval (CI): 3.64 6.64] in the Swedish study. These associations were confirmed in the U.S. population. Once a man's SNP genotypes and family history are known, his absolute risk for PCa can be readily calculated and easily interpreted. For example, 55 year-old men with a family history and >or=14 risk alleles have a 52% and 41% risk of being diagnosed with PCa in the next 20 years in the Swedish and U.S. populations, respectively. In comparison, without knowledge of genotype and family history, these men had an average population absolute risk of 13%. CONCLUSION: This risk prediction model may be used to identify men at considerably elevated PCa risk who may be selected for chemoprevention. PMID- 19562737 TI - Dynein-2 and ciliogenesis in Tetrahymena. AB - Dynein-2 is the motor responsible for retrograde intraflagellar transport. In situ, dynein-2 comprises four subunits: the dynein-2 heavy chain (DYH2); the dynein-2 intermediate chain; the dynein-2 light-intermediate chain (D2LIC); and dynein light chain 8 (Rompolas et al. 2007. Chlamydomonas FAP133 is a dynein intermediate chain associated with the retrograde intraflagellar transport motor. J Cell Sci 120:3653-3665). In contrast to what has been reported in other model organisms, when the DYH2 gene or the D2LIC gene was disrupted in Tetrahymena, the cells continued to produce motile cilia that were not swollen or filled with material [Rajagopalan et al.2009. Dynein-2 affects the regulation of ciliary length but is not required for ciliogenesis in Tetrahymena thermophila. Mol Biol Cell 20:708-720]. When compared to wildtype cells, the dynein-2 mutants were found to have cilia that were at a lower density, shorter, and much more variable in length. One possible explanation for these effects is that the dynein-2 knockout cells grow cilia too slowly to enable them to achieve normal length and density before the cell divides. In the present study, dynein-2 knockout cells were deciliated and then allowed to regrow their cilia for 22 hr under conditions in which the cells did not divide. When dynein-2 was disabled, three effects were observed: (1) a decreased rate of cilia growth; (2) a lower cilia density that did not change over time; and (3) a wide distribution of cilia lengths that increased over time. These results confirm the importance of dynein-2 in regulating ciliary length in Tetrahymena. Cell Motil. Cytoskeleton, 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19562738 TI - Tumour suppression by p53: the importance of apoptosis and cellular senescence. AB - p53 is regarded as a central player in tumour suppression, as it controls programmed cell death (apoptosis) as well as cellular senescence. While apoptosis eliminates cells at high risk for oncogenic transformation, senescence acts as a barrier to tumourigenesis by imposing irreversible cell cycle arrest. p53 can act directly or indirectly at multiple levels of the tumour suppression network by invoking a myriad of mechanisms. p53 induces the extrinsic and intrinsic apoptotic pathways at multiple steps to ensure an efficient death response. This response involves transcriptional activation or repression of target genes, as well as the recently identified microRNAs, and transcription-independent functions. Importantly, p53 loss of function is required for tumour maintenance. Therefore, therapeutic strategies aimed at reactivation of p53 in tumours emerge as a promising approach for the treatment of cancer patients. PMID- 19562739 TI - RUNX genes find a niche in stem cell biology. AB - The RUNX family of transcriptional regulators are well conserved throughout the animal kingdom, from the simple nematode worm Caenorhabditis elegans to vertebrates. Interest in the RUNX genes emerged principally as a result of the finding that chromosomal translocations disrupting RUNX protein function are observed in a large number of patients suffering with acute myeloid leukemia (AML). In the 20 years that RUNX genes have been under investigation, they have emerged as central players in the control of developmental decisions between proliferation and differentiation in a wide variety of biological situations. This review focuses on recent data highlighting the roles of RUNX genes in stem cells and illustrates the diversity of processes in which the RUNX proteins play a critical role. In particular, we focus on the role of RUNX1 in hematopoietic stem cells (HSCs) and hair follicle stem cells (HFSCs) and the importance of the solo C. elegans RUNX factor rnt-1 in stem cell proliferation in the worm. Observations in a variety of stem cell systems have developed to the point where useful comparisons can be made, from which guiding principles may emerge. PMID- 19562740 TI - SIRT1: a novel target to prevent atherosclerosis. AB - Atherosclerosis is a chronic immuno-inflammatory disease associated with blood lipids disorder. Many studies have demonstrated that caloric restriction (CR) can prevent atherosclerosis and extend lifespan. Sir2 protein, mammal's SIRT1, has been reported to at least partly contribute to the protective effect of CR. Hence, we hypothesize that SIRT1 is a key regulator in the pathogenesis of atherosclerosis and that upregulation of SIRT1 in endothelial cells may mimic CR's beneficial effect on vascular health. The recent studies have demonstrated that endothelial SIRT1 is an anti-atherosclerosis factor and the possible mechanism may be related to inhibit oxidized low-density lipoprotein (oxLDL) induced apoptosis, upregulate endothelial nitric oxide synthase (eNOS) expression, and improve endothelium relaxation function. We infer that SIRT1 may be a novel target for atherosclerosis prevention and treatment. PMID- 19562741 TI - Magnetic force microscopy of iron oxide nanoparticles and their cellular uptake. AB - Magnetic force microscopy has the capability to detect magnetic domains from a close distance, which can provide the magnetic force gradient image of the scanned samples and also simultaneously obtain atomic force microscope (AFM) topography image as well as AFM phase image. In this work, we demonstrate the use of magnetic force microscopy together with AFM topography and phase imaging for the characterization of magnetic iron oxide nanoparticles and their cellular uptake behavior with the MCF7 carcinoma breast epithelial cells. This method can provide useful information such as the magnetic responses of nanoparticles, nanoparticle spatial localization, cell morphology, and cell surface domains at the same time for better understanding magnetic nanoparticle-cell interaction. It would help to design magnetic-related new imaging, diagnostic and therapeutic methods. PMID- 19562742 TI - A novel electrospinning target to improve the yield of uniaxially aligned fibers. AB - Electrospinning is a useful technique that can generate micro and nanometer-sized fibers. Modification of the electrospinning parameters, such as deposition target geometry, can generate uniaxially aligned fibers for use in diverse applications ranging from tissue engineering to material fabrication. For example, meshes of fibers have been shown to mimic the extracellular matrix networks for use in smooth muscle cell proliferation. Further, aligned fibers can guide neurites to grow along the direction of the fibers. Here we present a novel electrospinning deposition target that combines the benefits of two previously reported electrodes: the standard parallel electrodes and the spinning wheel with a sharpened edge. This new target design significantly improves aligned fiber yield. Specifically, the target consists of two parallel aluminum plates with sharpened edges containing a bifurcating angle of 26 degrees. Electric field computations show a larger probable area of aligned electric field vectors. This new deposition target allows fibers to deposit on a larger cross-sectional area relative to the existing parallel electrode and at least doubles the yield of uniaxially aligned fibers. Further, fiber alignment and morphology are preserved after collection from the deposition target. PMID- 19562743 TI - Cytogenotoxicity induced by PBDE-47 combined with PCB153 treatment in SH-SY5Y cells. AB - Polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) are important recalcitrant halogenated compounds that have been regarded as major environmental pollutants. Recently, their concurrent appearance in the environment and humans and their structural and toxicological profile similarities have sparked interest in the potential toxicologic consequences of their coexposure. The aim of the current study was to evaluate the cytogenotoxic effects induced by 2,2',4,4'-tetrabromodiphenyl ether (PBDE-47) combined with 2,2',4,4',5,5'-hexachlorobiphenyl (PCB153) treatment in human neuroblastoma cells (SH-SY5Y) in vitro. SH-SY5Y cells were exposed to different concentrations of PBDE-47 (0, 2, 4, 8 MUM) with or without PCB153 (5 MUM) for 24 h. Thereafter, the cell viability, DNA damage, chromosomal abnormalities, and DNA-protein crosslinks (DPC) were determined. The results show that PBDE-47 and PCB153 alone and in combination induce DNA damage, with an increase in the frequency of micronuclei (MN) and DPC formation with increasing PBDE-47 concentration. In cells coexposed to PBDE-47 and PCB153, the cell viability significantly decreased while the MN frequency, DNA damage and DPC formation were all obviously increased compared to those of cells treated with the corresponding concentrations of PBDE-47 or PCB153 alone. Factorial analysis suggests that there were interactions between PBDE-47 and PCB153. The results imply that PBDE-47 interacts with PCB153 to inhibit cell viability and induce DNA damage, DPC formation, and chromosome abnormalities. PMID- 19562744 TI - The folliculin mutation database: an online database of mutations associated with Birt-Hogg-Dube syndrome. AB - The folliculin gene (FLCN), also known as BHD, is the only known susceptibility gene for Birt-Hogg-Dube syndrome. BHDS is the autosomal dominant predisposition to the development of follicular hamartomas, lung cysts, spontaneous pneumothorax, and/or kidney neoplasms. To date, 53 unique germline mutations have been reported. FLCN mutation detection rate is 88%. FLCN encodes a predicted 579 amino acid protein, designated folliculin that is highly conserved between humans and homologs in mice, Drosophila, and C. elegans. We developed the first online database detailing all FLCN variants identified in our laboratory and reported in the literature. The FLCN database applies, and assists researchers in applying HGVS nomenclature guidelines. To date, the FCLN database includes 84 variants: 53 unique germline mutations and 31 SNPs. The majority of FLCN germline mutations are predicted to produce a truncated folliculin, resulting in loss of function. The FLCN mutations consist of: 45% (24/53) deletions, 32% (17/53) substitutions (10 putative-splice site, 5 nonsense, and 2 missense), 15% (8/53) duplications, 6% (3/53) insertion/deletions and 2% (1/53) insertions. The database strives to systematically unify current knowledge of FLCN variants and will be useful to geneticists and genetic counselors while also providing a rapid and systematic resource for investigators. PMID- 19562745 TI - Liquid-based preparation for endometrial cytology--usefulness for predicting the prognosis of endometrial carcinoma preoperatively. AB - BACKGROUND: The authors evaluated the applicability and usefulness of immunocytochemical staining for cyclin A, p53, estrogen receptor alpha (ER alpha), and progesterone receptor B (PR-B) as a preoperative prognostic indicators for endometrial carcinoma using endometrial cytology with the liquid based cytology (LBC) method. METHODS: Cytologic specimens from 44 patients who had endometrial carcinoma were prepared with the LBC method. The results of immunocytochemical and immunohistochemical staining for cyclin A, p53, ER-alpha, and PR-B were compared with clinicopathologic parameters and prognosis. RESULTS: Patients who had positive results for cyclin A and p53 and negative results for ER-alpha and PR-B appeared to have unfavorable clinicopathologic characteristics, such as high-grade histology, advanced clinical stage, lymphovascular space involvement (LVSI), and deeper myometrial invasion (MI), and had a poor prognosis. In contrast, patients who had positive results for ER-alpha and PR-B, and negative results for cyclin A and p53 had favorable characteristics, such well differentiated tumor, early clinical stage, negative LVSI, and less MI, and had a good prognosis. Immunostaining results from cytologic specimens obtained in the clinic and at surgery and from histologic specimens obtained at surgery were correlated positively. CONCLUSIONS: Consistent specimens that were prepared using the LBC method facilitated multiple immunocytochemical analyses. Endometrial cytology with the LBC method was useful for predicting the prognosis of patients with endometrial carcinoma before therapy. PMID- 19562746 TI - Arg-85 and Thr-430 in murine 5-aminolevulinate synthase coordinate acyl-CoA binding and contribute to substrate specificity. AB - 5-Aminolevulinate synthase (ALAS) controls the rate-limiting step of heme biosynthesis in mammals by catalyzing the condensation of succinyl-coenzyme A and glycine to produce 5-aminolevulinate, coenzyme-A (CoA), and carbon dioxide. ALAS is a member of the alpha-oxoamine synthase family of pyridoxal 5'-phosphate (PLP) dependent enzymes and shares high degree of structural similarity and reaction mechanism with the other members of the family. The X-ray crystal structure of ALAS from Rhodobacter capsulatus reveals that the alkanoate component of succinyl CoA is coordinated by a conserved arginine and a threonine. The functions of the corresponding acyl-CoA-binding residues in murine erthyroid ALAS (R85 and T430) in relation to acyl-CoA binding and substrate discrimination were examined using site-directed mutagenesis and a series of CoA-derivatives. The catalytic efficiency of the R85L variant with octanoyl-CoA was 66-fold higher than that of the wild-type protein, supporting the proposal of this residue as key in discriminating substrate binding. Substitution of the acyl-CoA-binding residues with hydrophobic amino acids caused a ligand-induced negative dichroic band at 420 nm in the CD spectra, suggesting that these residues affect substrate mediated changes to the PLP microenvironment. Transient kinetic analyses of the R85K variant-catalyzed reactions confirm that this substitution decreases microscopic rates associated with formation and decay of a key reaction intermediate and show that the nature of the acyl-CoA tail seriously affect product binding. These results show that the bifurcate interaction of the carboxylate moiety of succinyl-CoA with R85 and T430 is an important determinant in ALAS function and may play a role in substrate specificity. PMID- 19562747 TI - In vitro response of monocyte-derived macrophages to a decellularized pericardial biomaterial. AB - Decellularized tissue-derived heart valves are an example of biomaterials derived from natural scaffolds. These types of implants are increasing in popularity although their in vivo performance is still only poorly understood and has, at times, been catastrophic. It is apparent that better understanding is required before these biomaterials can be used safely. In this study, the human monocyte derived macrophage (MDM) response to decellularized bovine pericardium (DBP) was used as a model to predict the biological performance of these materials on implantation. Human monocytes differentiated on tissue culture polystyrene (TCPS) for 14 days were trypsinized and reseeded onto DBP, TCPS, and polydimethylsiloxane (PDMS) for 48 h. The MDMs on DBP contained less intracellular and extracellular esterase activity compared with MDMs on TCPS and PDMS, as well as less acid phosphatase activity than on TCPS. As well, morphologically, MDMs on DBP were less spread, less multinucleated and did not display many lamellipodia. Taken together, these data represent the first evidence of the MDM response to intact, native extracellular matrix, demonstrating that these cells reacted with an altered, possibly reduced foreign body response on this natural scaffold compared with the two control surfaces. This in vitro MDM cell model may provide a novel method for predicting and elucidating the biological performance of tissue-derived biomaterials, thereby directing a more rational design of biomaterials for tissue regeneration purposes. PMID- 19562748 TI - FLT3 mutations have no prognostic impact in elderly patients with acute myeloid leukemia and normal karyotype. PMID- 19562749 TI - In vitro and in vivo evaluation on the bioactivity of ZnO containing nano hydroxyapatite/chitosan cement. AB - A ZnO containing nano-hydroxyapatite/chitosan (n-HA/CS) cement was developed and its bone formation ability was investigated in vitro and in vivo. The physico chemical properties of the cement were determined in terms of pH variation during and after setting, injectability and wettability. The results indicated that, the pH varied from 7.04 to 7.12 throughout the soaking of the cement in distilled water. The injectability was excellent during the first 4 min, but the cement became less injectable or even not injectable at all after 7 min setting. The static contact angle of the cement against water was 53.5 +/- 2.7 degrees . The results of immersion tests in simulated body fluid (SBF) indicated that the cement exhibited excellent bone-like apatite forming ability. In vivo studies, involving the installation of the cement of tibial-bone defects in rabbit tibia revealed an inflammatory response around the cement at 3 days of implantation. After 4 weeks, the inflammation began to disappear and the cement had bound to the surrounding host bone. Radiological examination also confirmed that the ZnO containing n-HA/CS cement significantly induced new bone formation. These results suggest that the ZnO containing n-HA/CS cement may be beneficial to enhance bone regeneration in osseous defect sites. PMID- 19562750 TI - The influence of surface energy on early adherent events of osteoblast on titanium substrates. AB - Surface energy of implant material is one of the important factors in the process of osseointegration. How surface energy regulates the signaling pathway of osteoblasts, however, is not well understood. Cell adhesion is one of the first steps essential to subsequent proliferation and differentiation of bone cells before tissue formation. Our present study was designed to investigate how surface energy may influence the early adhesion of human alveolar osteoblasts (AOBs). Substrates applied were two groups of titanium disks: (1) hydrophobic sandblasted and acid-etched (SLA) surfaces; (2) chemically modified hydrophilic SLA (modSLA) ones. Cell morphology and cell attachment were examined by scanning electron microscopy (SEM). Defined cytoskeletal actin organization was immunohistochemically examined using confocal laser scanning microscopy. RT-PCR was applied to detect and to compare the expression of focal adhesion kinase (FAK) of osteoblasts cultured on the two groups of substrates. The attachment rates of AOBs cultured on modSLA substrates were significantly higher than the cells on SLA ones within 3 h. AOBs on modSLA developed more defined actin stress fibers after 6 h of attachment. FAK expression was comparably higher on modSLA after 6 h. Within the limitation of the current study, higher surface energy of titanium surfaces enhanced the cell adhesion in the early stage of cell response and may work through influencing the expression of adhesion-associated molecules. PMID- 19562752 TI - Effect of CaTiO(3)-CaCO(3) prepared by alkoxide method on cell response. AB - In recent years, calcium titanate (CaTiO(3)) and carbon-containing materials have gained much attention in a number of biomedical material researches. To maximize the advantages of both materials, we developed a novel alkoxide method to get "calcium titanate with calcium carbonate" (CaTiO(3)-CaCO(3)). The objective was to evaluate the crystallinity and elemental composition of CaTiO(3)-CaCO(3) prepared by alkoxide method, CaTiO(3)-aC elaborated by modified thermal decomposition method, commercially-prepared CaTiO(3), and the effect of these materials on the bone marrow stromal cell. Hydroxyapatite was used as positive control material. We examined the cellular proliferation, osteoblastic differentiation, and mineralization of KUSA/A1 cells cultured with the materials. The results showed that CaTiO(3)-CaCO(3) and CaTiO(3)-aC contained evidence of calcium carbonate enhancing cell proliferation, osteoblastic differentiation, and mineralization. On the contrary, the commercially-prepared CaTiO(3) revealed absence of calcium carbonate with lower cell response than the other groups. The results indicated that calcium carbonate could play a key role in the cell response of CaTiO(3) material. In conclusion, our findings suggest that CaTiO(3) CaCO(3) could be considered an important candidate as a biomaterial for medical and dental applications. PMID- 19562755 TI - N-terminal truncated pyroglutamyl beta amyloid peptide Abetapy3-42 shows a faster aggregation kinetics than the full-length Abeta1-42. AB - We tested directly the differences in the aggregation kinetics of three important beta amyloid peptides, the full-length Abeta1-42, and the two N-terminal truncated and pyroglutamil modified Abetapy3-42 and Abetapy11-42 found in different relative concentrations in the brains in normal aging and in Alzheimer disease. By following the circular dichroism signal and the ThT fluorescence of the solution in phosphate buffer, we found substantially faster aggregation kinetics for Abetapy3-42. This behavior is due to the particular sequence of this peptide, which is also responsible for the specific oligomeric aggregation states, found by TEM, during the fibrillization process, which are very different from those of Abeta1-42, more prone to fibril formation. In addition, Abetapy3-42 is found here to have an inhibitory effect on Abeta1-42 fibrillogenesis, coherently with its known greater infective power. This is an indication of the important role of this peptide in the aggregation process of beta-peptides in Alzheimer disease. PMID- 19562754 TI - Centrosome amplification induced by the antiretroviral nucleoside reverse transcriptase inhibitors lamivudine, stavudine, and didanosine. AB - In cultured cells, exposure to the nucleoside reverse transcriptase inhibitor (NRTI) zidovudine (AZT) induces genomic instability, cell cycle arrest, micronuclei, sister chromatid exchanges, and shortened telomeres. In previous studies, we demonstrated AZT-induced centrosome amplification (>2 centrosomes/cell). Here, we investigate centrosome amplification in cells exposed to other commonly used NRTIs. Experiments were performed using Chinese Hamster ovary (CHO) cells, and two normal human mammary epithelial cell (NHMEC) strains: M99005 and M98040, which are high and low incorporators of AZT into DNA, respectively. Cells were exposed for 24 hr to lamivudine (3TC), stavudine (d4T), didanosine (ddI), and thymidine, and stained with anti-pericentrin antibody. Dose response curves were performed to determine cytotoxicity and a lower concentration at near plasma levels and a 10 fold higher concentration were chosen for the experiments. In CHO cells, there was a concentration-dependent, significant (P < 0.05) increase in centrosome amplification for each of the NRTIs. In NHMEC strain M99005, an NRTI-induced increase (P < 0.05) in centrosome amplification was observed for the high concentrations of each NRTI and the low doses of 3TC and ddI. In NHMEC strain M98040, the high doses of ddI and d4T showed significant increases in centrosome amplification. Functional viability of amplified centrosomes was assessed by arresting microtubule nucleation with nocodazole. In cells with more than two centrosomes, the ability to recover microtubule nucleation was similar to that of unexposed cells. We conclude that centrosome amplification is a consequence of exposure to NRTIs and that cells with centrosome amplification are able to accomplish cell division. PMID- 19562756 TI - Natural abundant solid state NMR studies in designed tripeptides for differentiation of multiple conformers. AB - Solid state NMR (SSNMR) experiments on heteronuclei in natural abundance are described for three synthetically designed tripeptides Piv-(L)Pro-(L)Pro-(L)Phe OMe (1), Piv-(D)Pro-(L)Pro-(L)Phe-OMe (2), and Piv-(D)Pro-(L)Pro-(L)Phe-NHMe (3). These peptides exist in different conformation as shown by solution state NMR and single crystal X-ray analysis (Chatterjee et al., Chem Eur J 2008, 14, 6192). In this study, SSNMR has been used to probe the conformations of these peptides in their powder form. The (13)C spectrum of peptide (1) showed doubling of resonances corresponding to cis/cis form, unlike in solution where the similar doubling is attributed to cis/trans form. This has been confirmed by the chemical shift differences of C(beta) and C(gamma) carbon of Proline in peptide (1) both in solution and SSNMR. Peptide (2) and (3) provided single set of resonances which represented all trans form across the di-Proline segment. The results are in agreement with the X-ray analysis. Solid state (15)N resonances, especially from Proline residues provided additional information, which is normally not observable in solution state NMR. (1)H chemical shifts are also obtained from a two-dimensional heteronuclear correlation experiment between (1)H--(13)C. The results confirm the utility of NMR as a useful tool for identifying different conformers in peptides in the solid state. PMID- 19562753 TI - Evolution of developmental regulation in the vertebrate FgfD subfamily. AB - Fibroblast growth factors (Fgfs) encode small signaling proteins that help regulate embryo patterning. Fgfs fall into seven families, including FgfD. Nonvertebrate chordates have a single FgfD gene; mammals have three (Fgf8, Fgf17, and Fgf18); and teleosts have six (fgf8a, fgf8b, fgf17, fgf18a, fgf18b, and fgf24). What are the evolutionary processes that led to the structural duplication and functional diversification of FgfD genes during vertebrate phylogeny? To study this question, we investigated conserved syntenies, patterns of gene expression, and the distribution of conserved noncoding elements (CNEs) in FgfD genes of stickleback and zebrafish, and compared them with data from cephalochordates, urochordates, and mammals. Genomic analysis suggests that Fgf8, Fgf17, Fgf18, and Fgf24 arose in two rounds of whole genome duplication at the base of the vertebrate radiation; that fgf8 and fgf18 duplications occurred at the base of the teleost radiation; and that Fgf24 is an ohnolog that was lost in the mammalian lineage. Expression analysis suggests that ancestral subfunctions partitioned between gene duplicates and points to the evolution of novel expression domains. Analysis of CNEs, at least some of which are candidate regulatory elements, suggests that ancestral CNEs partitioned between gene duplicates. These results help explain the evolutionary pathways by which the developmentally important family of FgfD molecules arose and the deduced principles that guided FgfD evolution are likely applicable to the evolution of developmental regulation in many vertebrate multigene families. PMID- 19562758 TI - Clonazepam is an effective treatment for hyperekplexia due to a SLC6A5 (GlyT2) mutation. PMID- 19562759 TI - Are current recommendations to diagnose orthostatic hypotension in Parkinson's disease satisfactory? AB - We interviewed 50 Parkinson's disease (PD) patients using a questionnaire to verify the reliability of orthostatic symptoms in warning the presence of orthostatic hypotension (OH). OH is defined as 20 mm Hg systolic or 10 mm Hg diastolic BP fall within 3 min of tilting or standing but if this fall occurs after 3 min we called it 'late OH' (L-OH). We compared if OH in Parkinson's disease (PD) was more frequent after head-up tilt or on standing and if the period of postural challenge matters in detecting OH. Twenty-one (42%) patients had OH that occurred twice more often after tilting (n = 20) than on standing (n = 10). OH occurred within 3 min of tilting in 9 patients (18%) and appeared beyond the currently recommended 3 min in 11 patients (55%) (L-OH). Ten of the 20 patients developing OH on tilting were symptomatic. The 10 patients who had OH on standing were asymptomatic. Reporting of symptoms was independent of age or severity of BP fall. Most (90%) patients reporting orthostatic symptoms on standing had OH on tilting for 3 min. Orthostatic symptoms in PD have a high specificity but low sensitivity in predicting OH. In Parkinson's disease OH occurs often after tilting than on standing and is delayed (after 3 min). As OH in PD is often asymptomatic and delayed it could contribute to falls and increase morbidity. We suggest routine evaluation of OH in PD by tilting them longer than the recommended 3 minutes to detect delayed OH. PMID- 19562760 TI - Embouchure dystonia--Portrait of a task-specific cranial dystonia. AB - Focal task-specific dystonia (FTSD) is an unusual disorder of motor control, which typically affects the hand but may also involve the face, jaw, and tongue. We report 89 musicians with dystonia of the embouchure (ED), the muscles of the lower face, jaw, and tongue used to control the flow of air into the mouthpiece of a woodwind or brass instrument. Symptoms of ED began at an average age of 36, were typically painless and only rarely were preceded by trauma. Specific musical techniques commonly triggered dystonia, often in one instrumental register. Task specific embouchure tremor and lip-pulling ED phenotypes were common among high register brass players (trumpet and French horn), whereas lip-locking occurred exclusively in low-register brass players (trombone and tuba). Jaw and tongue ED phenotypes occurred predominantly in woodwind players, and once present, frequently spread to speaking or eating. Six percent of all ED patients had coincident writer's cramp, suggesting a possible genetic predisposition to develop dystonia. We assessed two-point sensory discrimination in the upper lip, lower lip, and hand in ED patients, normal musicians, and nonmusician age-matched controls--there were no differences between groups. Once present, symptoms of ED did not remit and often disrupted careers and livelihoods. Better treatments are urgently needed for this unusual disorder of oral motor control. PMID- 19562762 TI - Norepinephrine and cardiovascular responses to maximal exercise in Parkinson's disease on and off medication. AB - The aim of this experiment is to understand how Parkinson's disease (PD) medication affects the autonomic responses of individuals during an acute exercise stress test. Fourteen people with PD and fifteen healthy individuals age matched between 50 and 80 years performed a modified Bruce protocol. Subjects with PD performed the test once off medication (PD-off) and then 1 week later on medication (PD-on). Heart rate (HR), blood pressure (BP), VO(2), and norepinephrine (NE) levels were taken at rest and at peak exercise. At peak exercise HR, BP, and NE values for the PD-on and PD-off group were all significantly lower than healthy controls, regardless of whether subjects were on their medication. Autonomic abnormalities during exercise in this population appear to be disease manifested and not impacted by medications used to treat PD. We can assume, both on and off medication, this population will show markedly lower BP, HR, and NE responses. PMID- 19562763 TI - GIGYF2 variants are not associated with Parkinson's disease in Italy. PMID- 19562765 TI - Paraneoplastic movement disorders. AB - Paraneoplastic movement disorders are rare autoimmune nonmetastatic complications of cancer. Common paraneoplastic movement disorders include cerebellar syndrome, opsoclonus myoclonus, basal ganglia disorders, stiff person syndrome, and neuromyotonia. Syndromes usually present before cancer diagnosis and are commonly associated with one or more serum antibodies. Increasing numbers of antibodies have been identified (Hu, Yo, Ri, CV2, amphiphysin, Ma, Ta, Tr, NMDA, mGluR1, PCA2, ANNA-3, VGCCA). Antibodies are highly correlated with the likelihood of an underlying cancer and are closely associated with certain tumors. Clinical clues to paraneoplastic aetiology include speed of onset, severity, speed of progression, resistance to treatment, and more widespread neurological signs than one would expect from nonparaneoplastic aetiologies. Cancer should be sought in those with classical presentations and those with possible presentations who have paraneoplastic antibodies. If no tumor is found on initial investigation, interval screening is advisable. The most common associated cancers found are small cell lung cancer, breast, gynaecological, testicular, lymphoma, and thymoma. Early identification and treatment sometimes leads to neurological improvement and may improve cancer prognosis. Prognosis is dependent on the tumor type and its likely response to treatment. PMID- 19562767 TI - Paraneoplastic cataplexy: clinical presentation and imaging findings in a case. PMID- 19562764 TI - The morphology and spatial arrangement of astrocytes in the optic nerve head of the mouse. AB - We evaluated the shapes, numbers, and spatial distribution of astrocytes within the glial lamina, an astrocyte-rich region at the junction of the retina and optic nerve. A primary aim was to determine how the population of astrocytes, collectively, partitions the axonal space in this region. Astrocyte processes labeled with glial fibrillary acidic protein (GFAP) compartmentalize ganglion cell axons into bundles, forming "glial tubes," and giving the glial architecture of the optic nerve head in transverse section a honeycomb appearance. The shapes of individual astrocytes were studied by using transgenic mice that express enhanced green fluorescent protein in isolated astrocytes (hGFAPpr-EGFP). Within the glial lamina the astrocytes were transverse in orientation, with thick, smooth primary processes emanating from a cytoplasmic expansion of the soma. Spaces between the processes of neighboring astrocytes were spatially aligned, to form the apertures through which the bundles of optic axons pass. The processes of individual astrocytes were far-reaching-they could span most of the width of the nerve-and overlapped the anatomical domains of other near and distant astrocytes. Thus, astrocytes in the glial lamina do not tile: each astrocyte participates in ensheathing approximately one-quarter of all of the axon bundles in the nerve, and each glial tube contains the processes of about nine astrocytes. This raises the mechanistic question of how, in glaucoma or other cases of nerve damage, the glial response can be confined to a circumscribed region where damage to axons has occurred. PMID- 19562766 TI - Comparison of three clinical rating scales in Friedreich ataxia (FRDA). AB - To test the validity and reliability of the scale for the assessment and rating of ataxia (SARA) in Friedreich ataxia (FRDA). SARA is limited to eight items and can be performed rapidly. Ninety-six patients with a molecular genetic diagnosis of FRDA were rated using three different clinical scales, the FRDA Rating Scale (FARS), the International Cooperative Ataxia Rating Scale (ICARS), and SARA. Despite considerable discrepancies in scale size and subscale structure, SARA total scores were significantly correlated with ICARS (r = 0.953, P < 0.0001) and FARS (r = 0.938, P < 0.0001) total scores. SARA total scores also correlated with the activities of daily living (ADL, r = 0.929, P < 0.0001). Although originally developed for the use in dominantly inherited ataxias, which are primarily ataxias of the cerebellar type, SARA can also be used successfully to assess afferent ataxia, which is the predominant form in FRDA. Because SARA is characterized by high interrater reliability and practicability, SARA is applicable and well suited forclinical trials of FRDA. PMID- 19562761 TI - Motor abnormalities in premanifest persons with Huntington's disease: the PREDICT HD study. AB - The PREDICT-HD study seeks to identify clinical and biological markers of Huntington's disease in premanifest individuals who have undergone predictive genetic testing. We compared baseline motor data between gene-expansion carriers (cases) and nongene-expansion carriers (controls) using t-tests and Chi-square. Cases were categorized as near, mid, or far from diagnosis using a CAG-based formula. Striatal volumes were calculated using volumetric magnetic resonance imaging measurements. Multiple linear regression associated total motor score, motor domains, and individual motor items with estimated diagnosis and striatal volumes. Elevated total motor scores at baseline were associated with higher genetic probability of disease diagnosis in the near future (partial R(2) 0.14, P < 0.0001) and smaller striatal volumes (partial R(2) 0.15, P < 0.0001). Nearly all motor domain scores showed greater abnormality with increasing proximity to diagnosis, although bradykinesia and chorea were most highly associated with diagnostic immediacy. Among individual motor items, worse scores on finger tapping, tandem gait, Luria, saccade initiation, and chorea show unique association with diagnosis probability. Even in this premanifest population, subtle motor abnormalities were associated with a higher probability of disease diagnosis and smaller striatal volumes. Longitudinal assessment will help inform whether motor items will be useful measures in preventive clinical trials. PMID- 19562768 TI - The W cell pathway to cat primary visual cortex. AB - The thalamic input to area 17 in the cat can be divided into at least three parallel pathways, the W, X, and Y. Although the latter two are some of the best studied synaptic connections in the brain, the former remains poorly understood both in structure and in function. By combining light and electron microscopy, we have reconstructed in 3-D single W axons and described quantitatively the synapses that they form. We have also made a structural comparison of reconstructed synapses from the three visual pathways. Thalamic axons were labeled in vivo by injections of biotinylated dextran amine into the dLGN. W axons originating from C laminae injections arborized in layers 1, 2/3, and 5. Axons that traversed layer 1 supplied a few descending collaterals to layer 2/3, but the most extensive innervation in layer 2/3 was provided by axons ascending from the white matter. Most W boutons formed a single synapse, dendritic spines being the most common target, with dendritic shafts forming the remaining targets. In layer 1, the area of the postsynaptic density of spine synapses (0.16 microm(2)) was significantly larger than that of layers 2/3 (0.11 microm(2)) and 5 (0.09 microm(2)). Synapses from X and Y axons in layer 4 were similar in size to synapses formed by W boutons in layer 1. In layer 1, the main targets of the W axons are likely the apical dendrites of pyramidal cells, so that both proximal and distal regions of pyramidal cell dendritic trees can be excited by the W pathway. PMID- 19562769 TI - Association of DRD3 and GRIN2B with impulse control and related behaviors in Parkinson's disease. AB - We aimed to assess whether allelic variants of dopamine receptor, glutamate receptor, and serotonin transporter genes are associated with the appearance of impulse control and related behaviors (ICRB) in Parkinson's disease (PD) with dopamine replacement therapy (DRT). We surveyed ICRB in consecutive Korean patients with PD who were treated with stable DRT using modified Minnesota Impulsive Disorders Interview over a period of 4 months. In the 404 patients who completed the interview and the 559 Korean healthy normal controls, genotyping was performed for variants of the DRD3 p.S9G, DRD2 Taq1A, GRIN2B c.366C>G, c.2664C>T and c.-200T>G, and the promoter region of the serotonin transporter gene (5-HTTLPR). Behavioral abnormalities suggestive of ICRB including compulsive buying, gambling, sexual behavior and eating, and punding, were present in 14.4% of the patients. Variants of DRD2 and 5-HTTLPR were not associated with the risk of developing ICRB. However, the AA genotype of DRD3 p.S9G and the CC genotype of GRIN2B c.366C>G were more frequent in patients with ICRB than in nonaffected patients (odds ratio [OR] = 2.21, P = 0.0094; and 2.14, P = 0.0087, after adjusting for age and sex). After controlling for clinical variables in the multivariate analysis, carriage of either AA genotype of DRD3 or CC genotype of GRIN2B was identified as an independent risk factor for ICRB (adjusted OR: 2.57, P = 0.0087). Variants of DRD3 p.S9G and GRIN2B c.366C>G may be associated with the appearance of ICRB in PD. PMID- 19562771 TI - Cervical dystonia and Constantin Brancusi. PMID- 19562770 TI - Expanding the clinical phenotype of SNCA duplication carriers. AB - SNCA duplication is a recognized cause of familial Parkinson's disease (PD). We aimed to explore the genetic and clinical variability in the disease manifestation. Molecular characterization was performed using real-time PCR, SNP arrays, and haplotype analysis. We further studied those patients who were found to harbor SNCA duplication with olfactory function tests, polysomnography, and PET. We identified four new families and one sporadic patient with SNCA duplication. Eleven symptomatic patients from these four families presented with parkinsonism, of which three subsequently developed dementia. The lifetime estimate of overall penetrance was 43.8%. FDG-PET study of symptomatic patients showed hypometabolism in the occipital lobe, whereas asymptomatic carriers of SNCA duplication demonstrated normal glucose metabolism. Symptomatic patients showed abnormal olfactory function and polysomnography and asymptomatic carriers showed normal results. The clinical features of SNCA duplication include parkinsonism with or without dementia. Asymptomatic carriers displayed normal test results with the eldest individual aged 79 years; thus, even a carrier of SNCA duplication may escape the development of PD. This difference in age associated penetrance may be due to the genetic background or environmental exposures. Further studies of SNCA duplication carriers will help identify disease-modifiers and may open novel avenues for future treatment. PMID- 19562772 TI - Pulmonary neuroendocrine/carcinoid tumors: a review article. AB - Neuroendocrine tumors are a unique malignant neoplasm that can arise from the respiratory tree. Although well-differentiated bronchial neuroendocrine tumors (also called carcinoid tumors) are reported to account for approximately 25% of all neuroendocrine tumors, they represent only 1% to 2% of all lung cancers. The epidemiology, clinical behavior, and treatment of neuroendocrine carcinoid tumors differ significantly from other lung malignancies. In this article, the recent data regarding these tumors were reviewed with attention to the treatment modalities used. Although conventional cytotoxic therapy has not been reported to demonstrate much promise in this entity over the past 4 decades, newer molecular targeted agents including those that targeted angiogenesis and the mammalian target of rapamycin (mTOR) pathway have shown encouraging results in early phase trials for advanced carcinoid tumors. PMID- 19562773 TI - Development of a brief ataxia rating scale (BARS) based on a modified form of the ICARS. AB - To develop a brief ataxia rating scale (BARS) for use by movement disorder specialists and general neurologists. Current ataxia rating scales are cumbersome and not designed for clinical practice. We first modified the International Cooperative Ataxia Rating Scale (ICARS) by adding seven ataxia tests (modified ICARS, or MICARS), and observed only minimally increased scores. We then used the statistics package R to find a five-test subset in MICARS that would correlate best with the total MICARS score. This was accomplished first without constraints and then with the clinical constraint requiring one test each of Gait, Kinetic Function-Arm, Kinetic Function-Leg, Speech, and Eye Movements. We validated these clinical constraints by factor analysis. We then validated the results in a second cohort of patients; evaluated inter-rater reliability in a third cohort; and used the same data set to compare BARS with the Scale for the Assessment and Rating of Ataxia (SARA). Correlation of ICARS with the seven additional tests that when added to ICARS form MICARS was 0.88. There were 31,481 five-test subtests (48% of possible combinations) that had a correlation with total MICARS score of > or =0.90. The strongest correlation of an unconstrained five-test subset was 0.963. The clinically constrained subtest validated by factor analysis, BARS, had a correlation with MICARS-minus-BARS of 0.952. Cronbach alpha for BARS and SARA was 0.90 and 0.92 respectively; and inter-rater reliability (intraclass correlation coefficient) was 0.91 and 0.93 respectively. BARS is valid, reliable, and sufficiently fast and accurate for clinical purposes. PMID- 19562774 TI - Pallidal stimulation for segmental dystonia: long term follow up of 11 consecutive patients. AB - Pallidal stimulation is a convincing and valid alternative for primary generalized dystonia refractory to medical therapy or botulinum toxin. However, the clinical outcome reported in literature is variable most likely because of heterogeneity DBS techniques employed and /or to clinical dystonic pattern of the patients who undergo surgery. In this study, we report the long term follow up of a homogeneous group of eleven subjects affected by segmental dystonia who were treated with bilateral stimulation of the Globus Pallidus pars interna (GPi) from the years 2000 to 2008. All the patients were evaluated, before surgery and at 6 12-24-36 months after the treatment, in accordance with the Burke Fahn Marsden Dystonia Rating Scale (BFMDRS). Our study indicates that DBS promotes an early and significant improvement at 6 months with an even and a better outcome later on. The analysis of specific sub items of the BFMDRS revealed an earlier and striking benefit not only as far as segmental motor function of the limbs but also for the complex cranial functions like face, (eyes and mouth), speech and swallowing, differently from results reported in primary generalized dystonia. Deep Brain Stimulation of GPi should be considered a valid indication for both generalized and segmental dystonia when other therapies appear ineffective. PMID- 19562775 TI - PINK1 mutations in a Brazilian cohort of early-onset Parkinson's disease patients. AB - Data on the frequency of PINK1 mutations in Brazilian patients with early-onset Parkinson's disease (EOPD) are lacking. The aim of this report was to investigate mutations of the PINK1 gene in a cohort of Brazilian patients with EOPD. Sixty consecutive familial or sporadic EOPD patients were included. All eight PINK1 exons and exon-intron boundaries were analyzed. We did not find any pathogenic mutation of PINK1 in our cohort. Single Nucleotide Polymorphisms (SNP) were identified in 46.7% of the patients and in 45.9% of controls (P = 0.9). The SNPs identified in our patients had already been described in previous reports. The results of our study support the hypothesis that mutations in PINK1 may not be a relevant cause of EOPD. In Brazil, if we consider only EOPD patients, it seems that parkin and LRRK2 mutations are more common. PMID- 19562776 TI - A comprehensive geriatric assessment is more effective than clinical judgment to identify elderly diffuse large cell lymphoma patients who benefit from aggressive therapy. AB - BACKGROUND: The authors set out to analyze if a simple comprehensive geriatric assessment (CGA) could objectively identify elderly patients with diffuse large cell lymphoma (DLCL) who can be effectively treated with anthracycline-containing immunochemotherapy. METHODS: CGA was performed in 84 consecutive patients with DLCL aged >65 years and diagnosed at a single institution. Treatment with curative versus palliative intent was chosen according to clinical judgment. Cyclophosphamide, hydroxydaunomycin, Oncovin (vincristine), and prednisone (CHOP) or CHOP-like regimens were given to 62 (74%) patients. The outcome of patients was analyzed according to both the treatment received and the results of CGA. RESULTS: According to CGA, 42 (50%) patients were classified as "fit." They were younger (P < .0001) and had less frequent systemic symptoms (P = .03). These patients received curative treatment by clinical judgment. Their response rate (92.5% vs 48.8%; P < .0001) and median survival (not reached vs 8 months; P < .0001) were significantly better than those of 42 patients considered "unfit" by CGA. Among unfit patients, 20 had actually received curative and 22 palliative therapy. These subgroups did not differ in any geriatric or lymphoma-related characteristic. Their outcome was similar irrespectively of the type of treatment received (median survival, 8 vs 7 months; P = nonsignificant). Lymphoma rather than toxicity was the main cause of failure/death also among unfit patients treated aggressively. CONCLUSIONS: CGA is an efficient method to identify elderly DLCL patients who can benefit from a curative approach with anthracycline containing immunochemotherapy. Further study is needed to discern why unfit patients seem to have poor outcomes because of poor tolerance but also because of lymphoma refractoriness to intensive therapy. PMID- 19562777 TI - Clinical trial enrollment, patient characteristics, and survival differences in prospectively registered metastatic colorectal cancer patients. AB - BACKGROUND: Trial accrual patterns were examined to determine whether metastatic colorectal cancer (mCRC) patients enrolled in trials are representative of a general cancer population concerning patient characteristics and survival. METHODS: A total of 760 mCRC patients referred for their first oncological consideration at 3 hospitals in Scandinavia covering defined populations were registered consecutively during 2003 to 2006. Clinical trial enrollment, patient characteristics, and treatment were recorded prospectively, and the follow-up was complete. RESULTS: Palliative chemotherapy was initiated in 61% of the patients. Approximately one-third (36%) of patients receiving chemotherapy were included in a trial. The main reason for nonparticipation was failed eligibility criteria (69%). The median survival after chemotherapy was 15.8 months for all patients, and 18 months after combination chemotherapy. Trial patients had better prognostic characteristics and significantly longer survival than nontrial patients: 21.3 months versus 15.2 months when receiving combination chemotherapy. Poor performance status was the main reason for giving best supportive care only, and the median survival was then only 2.1 months. The median survival for all 760 nonresectable mCRC patients was 10.7 months. CONCLUSIONS: mCRC patients enrolled into clinical trials differ in characteristics from patients receiving chemotherapy outside protocol and have better survival, even when given the same treatment. Although trial patients have a median survival close to 2 years, survival is lower for all patients receiving chemotherapy and much lower for all patients diagnosed with mCRC. Studies that better accept the heterogeneity of the population with mCRC are needed. PMID- 19562779 TI - Cognitive behavioral therapy for psychogenic movement disorder. PMID- 19562780 TI - Association of age and survival in patients with gastroesophageal cancer undergoing surgery with or without preoperative therapy. AB - BACKGROUND: Meticulous selection of patients for esophageal cancer surgery is critical, because major surgical intervention can cause considerable consequences. For this study, the authors explored their institution's large surgical experience to examine the impact of age on long-term patient survival and surgical complications. METHOD: Six hundred consecutive patients with esophageal cancer who underwent surgery (409 patients received preoperative therapy, and 191 patients underwent surgery first) were analyzed. All demographic information (including American Society of Anesthesiology risk scores) and therapy-related information was collected retrospectively. Multiple statistical methods were used to assess survival rates and surgical complications and their correlation with patient age. Twenty-one patients (30-day mortality) first were excluded (n = 600) and then were included (n = 621) in the analysis. RESULTS: By using the median age (70 years) in patients older than the median age, univariate analysis demonstrated a higher risk of death with increasing age (P = .019). In multivariate analysis, increasing age was an independent prognosticator of poor overall survival (P = .041). The inclusion of 30-day mortality did not alter the results. Surgical complications were statistically significantly higher in older patients compared with younger patients in the following categories: aspiration pneumonia, adult respiratory distress syndrome, cardiovascular, neurologic, and miscellaneous complications. CONCLUSIONS: The data in this study demonstrated that patients aged 70 years. Age was identified as an important variable in the selection of patients for esophageal cancer surgery. PMID- 19562778 TI - Wnt antagonist gene polymorphisms and renal cancer. AB - BACKGROUND: Epigenetic silencing of several wingless-type mouse mammary tumor virus integration site (Wnt) pathway-related genes has been reported in renal cancer. Except for the T-cell factor 4 gene TCF4, there are no reports regarding Wnt pathway gene polymorphisms in renal cancer. Therefore, the authors of this report hypothesized that the polymorphisms in Wnt signaling genes may be risk factors for renal cancer. METHODS: In total, 210 patients (145 men and 65 women) with pathologically confirmed renal cell carcinoma (RCC) and 200 age-matched and sex-matched control individuals were enrolled in this study. We genotyped 14 single nucleotide polymorphisms (SNPs) in 6 genes. including Dickkopf 2 (DKK2) (reference SNP identification number 17037102 [rs17037102], rs419558, and rs447372), DKK3 (rs3206824, rs11022095, rs1472189, rs7396187, and rs2291599), DKK4 (rs2073664), secreted frizzled-related protein 4 (sFRP4) (rs1802073 and rs1802074), mothers against decapentaplegic homolog (SMAD) family member 7 or SMAD7 (rs12953717), and disheveled associated activator of morphogenesis 2 or DAAM2 (rs6937133 and rs2504106) using polymerase chain reaction-restriction fragment length polymorphism analysis and direct sequencing in the patients with RCC and in the healthy, age-matched control group. The relations also were tested between these polymorphisms and clinicopathologic data, including sex, tumor grade, tumor stage, lymph node involvement, distant metastasis, and overall survival. RESULTS: A significant decrease in the frequency of the guanine/adenine (G/A) + A/A genotypes in the DKK3 codon 335 rs3206824 was observed in the patients with RCC compared with the control group. The frequency of the rs3206824 (G/A) A-rs7396187 (guanine/cytosine [G/C]) C haplotype was significantly lower in patients with RCC compared with other haplotypes. In addition, DKK3 rs1472189 cytosine/thymine (C/T) was associated with distant metastasis, and, DKK2 rs17037102 G-homozygous patients had a decreased risk for death in multivariate Cox regression analysis. CONCLUSIONS: To the authors' knowledge, this is the first report documenting that DKK3 polymorphisms are associated with RCC and that the DKK2 rs17037102 polymorphism may be a predictor for survival in patients with RCC after radical nephrectomy. PMID- 19562781 TI - Release of toxic Gd3+ ions to tumour cells by vitamin B12 bioconjugates. AB - Two probes consisting of vitamin B(12) (CNCbl) conjugated to Gd chelates by esterification of the ribose 5'-OH moiety, Gd-DTPA-CNCbl (1; DTPA = diethylenetriamine-N,N,N',N'',N''-pentaacetic acid) and Gd-TTHA-CNCbl (2; TTHA = triethylenetetramine-N,N,N',N'',N''',N'''-hexaacetic acid), have been synthesised and characterised. The crystal structure of a dimeric form of 1, obtained by crystallisation with an excess of GdCl(3), has been determined. The kinetics of binding to and dissociation from transcobalamin II show that 1 and 2 maintain high-affinity binding to the vitamin B(12) transport protein. Complex 2 is very stable with respect to Gd(3+) release owing to the saturated co-ordination of the Gd(3+) ion by four amino and five carboxylate groups. Hydrolysis of the ester functionality occurs on the time scale of several hours. The lack of saturation and the possible involvement of the ester functionality in co-ordination result in lower stability of 1 towards hydrolysis and in a considerable release of Gd(3+) in vitro. Gd(3+) ions released from 1 are avidly taken up by the K562 tumour cells to an extent corresponding to approximately 10(10) Gd(3+) per cell. The internalisation of toxic Gd(3+) ions causes a marked decrease in cell viability as assessed by Trypan blue and WST-1 tests. On the contrary, the experiments with the more stable 2 did not show any significant cell internalisation of Gd(3+) ions and any influence on cell viability. The results point to new avenues of in situ generation of cytotoxic pathways based on the release of toxic Gd(3+) ions by vitamin B(12) bioconjugates. PMID- 19562782 TI - The controlled formation and cleavage of an intramolecular d8-d8 Pt-Pt interaction in a dinuclear cycloplatinated molecular "pivot-hinge". AB - The bis(diphenylphosphino)methane (dppm)-bridged dinuclear cycloplatinated complex {[Pt(L)](2)(mu-dppm)}(2+) (Pt(2)dppm; HL: 2-phenyl-6-(1H-pyrazol-3-yl) pyridine) demonstrates interesting reversible "pivot-hinge"-like intramolecular motions in response to the protonation/deprotonation of L. In its protonated "closed" configuration, the two platinum(II) centers are held in position by intramolecular d(8)-d(8) Pt-Pt interaction. In its deprotonated "open" configuration, such Pt-Pt interaction is cleaved. To further understand the mechanism behind this hingelike motion, an analogous dinuclear cycloplatinated complex, {[Pt(L)](2)(mu-dchpm)}(2+) (Pt(2)dchpm) with bis(dicyclohexylphosphino)methane (dchpm) as the bridging ligand, was synthesized. From its protonation/deprotonation responses, it was revealed that aromatic pi-pi interactions between the phenyl moieties of the mu-dppm and the deprotonated pyrazolyl rings of L was essential to the reversible cleavage of the intramolecular Pt-Pt interaction in Pt(2)dppm. In the case of Pt(2)dchpm, spectroscopic and spectrofluorometric titrations as well as X-ray crystallography indicated that the distance between the two platinum(II) centers shrank upon deprotonation, thus causing a redshift in its room-temperature triplet metal metal-to-ligand charge-transfer emission from 614 to 625 nm. Ab initio calculations revealed the presence of intramolecular hydrogen bonding between the deprotonated and negatively charged 1-pyrazolyl-N moiety and the methylene CH and phenyl C-H of the mu-dppm. The "open" configuration of the deprotonated Pt(2)dppm was estimated to be 19 kcal mol(-1) more stable than its alternative "closed" configuration. On the other hand, the open configuration of the deprotonated Pt(2)dchpm was 6 kcal mol(-1) less stable than its alternative closed configuration. PMID- 19562783 TI - A study of the influence of ionic liquids properties on the Kemp elimination reaction. AB - The morpholino-induced elimination of 5-nitrobenzisoxazole into the relevant 2 cyano-4-nitrophenolate has been used as a sample reaction in order to investigate molecular properties of some room temperature ionic liquids. The kinetic study was carried out at 298 K by means of spectrophotometric measurements. Ionic liquids, which differ in both their cation and anion properties, were used as solvent systems. In particular, aliphatic (pyrrolidinium, piperidinium, and ammonium) and aromatic (imidazolium and pyridinium) ionic liquids were used. For aromatic cations, imidazolium ions having different hydrogen-bond donor ability or a different alkyl-chain length were taken into account. The anions chosen ([BF(4)(-)], [PF(6)(-)], [SbF(6)(-)], and [NTf(2)(-)]; where NTf(2) = bis(trifluoromethansulfonyl)imide) showed different shape, size, and coordination ability. Solvent parameters of all ionic liquids used were determined by using spectroscopic probes, such as 4-nitroaniline, N,N-diethyl-4-nitroaniline, Nile Red, and Reichardt's dye. Finally, in order to obtain information on the structural organization of the solvent systems, resonance light-scattering measurements were carried out. The collected data provide evidence that ionic liquids are solvent media which exhibit peculiar features, whose effects can be rationalized only considering all parameters affecting their three-dimensional structure. PMID- 19562784 TI - Is cyclopropane really the sigma-aromatic paradigm? AB - Dewar proposed the sigma-aromaticity concept to explain the seemingly anomalous energetic and magnetic behavior of cyclopropane in 1979. While a detailed, but indirect energetic evaluation in 1986 raised doubts-"There is no need to involve 'sigma-aromaticity',"-other analyses, also indirect, resulted in wide-ranging estimates of the sigma-aromatic stabilization energy. Moreover, the aromatic character of "in-plane", "double", and cyclically delocalized sigma-electron systems now seems well established in many types of molecules. Nevertheless, the most recent analysis of the magnetic properties of cyclopropane (S. Pelloni, P. Lazzeretti, R. Zanasi, J. Phys. Chem. A 2007, 111, 8163-8169) challenged the existence of an induced sigma-ring current, and provided alternative explanations for the abnormal magnetic behavior. Likewise, the present study, which evaluates the sigma-aromatic stabilization of cyclopropane directly for the first time, fails to find evidence for a significant energetic effect. According to ab initio valence bond (VB) computations at the VBSCF/cc-PVTZ level, the sigma-aromatic stabilization energy of cyclopropane is, at most, 3.5 kcal mol(-1) relative to propane, and is close to zero when n-butane is used as reference. Trisilacyclopropane also has very little sigma-aromatic stabilization, compared to Si(3)H(8) (6.3 kcal mol(-1)) and Si(4)H(10) (4.2 kcal mol(-1)). Alternative interpretations of the energetic behavior of cyclopropane (and of cyclobutane, as well as their silicon counterparts) are supported. PMID- 19562785 TI - Lanthanide(III) complexes with two hexapeptides incorporating unnatural chelating amino acids: secondary structure and stability. AB - Unnatural metal-chelating amino acids bearing aminodiacetate side-chains have been introduced into two hexapeptides to obtain efficient lanthanide-binding peptides. The synthesis of the enantiopure Fmoc-Ada(n)(tBu)2-OH synthons is described with overall yields of 32 and 50% for n=2 and n=3 side-chain carbon atoms, respectively. The two peptides AcWAda(n)PGAda(n)GNH2 (Pn) were synthesized from the protected synthons by standard solid-phase peptide synthesis. Studies of the lanthanide complexes of the two peptides Pn by luminescence titrations, mass spectrometry, circular dichroism, and solution NMR spectroscopy demonstrate that the Ada(n) chain length has a dramatic effect on the complexation properties. Indeed, the flexible compound P3 forms a mononuclear complex of moderate stability (beta11=10(9.9)), which tends to transform into a binuclear species in the presence of excess of the metal ion. Interestingly, the more compact peptide P2 provides stable Ln3+ complexes with the exclusive formation of the mononuclear LnP2 adduct. The stability constant of TbP2 is two orders of magnitude higher (beta11=10(12.1)) than that measured for P3. The 800 MHz NMR spectrum of the La3+ complex of P2 evidences a well-defined type II beta-turn as well as a hydrophobic Trp(indole)-Pro interaction. These interactions exemplify the non-innocent character of the peptide spacer in the complex LaP2 as well as the role of a peptide secondary structure in the stabilization of metal complexes. PMID- 19562786 TI - Probing lanthanide anisotropy in Fe-Ln aggregates by using magnetic susceptibility measurements and 57Fe Mossbauer spectroscopy. PMID- 19562787 TI - A straightforward total synthesis of (-)-chaetominine. AB - A total synthesis of the tripeptide alkaloid (-)-chaetominine (1) was achieved in 9.3% overall yield starting from commercially available D-tryptophan methyl ester, based on a short and straightforward (nine steps) sequence. The early stage introduction (first step) of the quinazolinone moiety and the late stage introduction (penultimate step) of the hydroxy group allowed a synthetic strategy devoid of protective groups. The key step of the process is the a-c tricyclic ring construction via an unprecedented NCS-mediated N-acyl cyclization on an indole ring to give tetrahydro-1H-pyrido[2,3-b]indole 11. In the penultimate step, oxidation of the tetracyclic intermediate 14 with oxaziridine 15 gave only one of the four possible diastereoisomers, the cis-diastereoisomer 16 resulting from the attack of the oxaziridine to the double bond face opposite to the c-d ring substituents. In the last step, the complete stereocontrol of the Et(3)SiH/TFA reduction of compound 16, probably involving a N-acyliminium ion, can be attributed to ring constrain, which forces the b-c ring junction in the more stable cis-orientation. (-)-Chaetominine (1) showed a negligible inhibitory activity on several cancer cell lines. PMID- 19562788 TI - The Synechocystis sp. PCC6803 Sfp-type phosphopantetheinyl transferase does not possess characteristic broad-range activity. AB - The cyanobacterium Synechocystis sp. PCC6803 harbours one phosphopantetheinyl transferase (PPTase), Sppt. Protein modelling supported previous bioinformatics analyses, which suggested that Sppt is a Sfp-type PPTase with the potential to phosphopantetheinylate a broad range of carrier proteins from both primary and secondary metabolism. However, no natural products are synthesised by this species, which raises interesting evolutionary and functional questions. Phosphopantetheinylation assays and kinetic data demonstrate that Sppt was able to activate its cognate fatty acid synthesis carrier protein, SACP, but was unable to effectively activate various cyanobacterial carrier proteins from secondary metabolism or glycolipid biosynthesis pathways. To our knowledge, this is the first example of a PPTase with a Sfp-type structure, but with activity more closely resembling AcpS-type enzymes. The broad-range PPTase from Nodularia spumigena NSOR10 was introduced into Synechocystis sp. PCC6803 and was shown to activate a noncognate carrier protein, in vivo. This engineered strain could provide a future biotechnological platform for the heterologous expression of cyanobacterial biosynthetic gene clusters. PMID- 19562790 TI - Zeolite-catalyzed isomerization of triose sugars. PMID- 19562791 TI - Oxygen-ion transport in a dual-phase scandia-yttria-stabilized zirconia solid electrolyte: a molecular dynamics simulation. AB - A molecular dynamics (MD) simulation is adopted to investigate oxygen-ion diffusion mechanisms in dual-phase (cubic/monoclinic phase) zirconia-based solid electrolytes. XRD analysis is performed to validate the phase structure composition after an MD duration of 500 ps. The radial distribution function of the ion pairs is used to analyze the microstructure and mobility of the oxygen ions inside the electrolyte. The mean-squared displacement, displacement distribution, and moving path of oxygen ions indicate that ion mobility increases with Sc(2)O(3) content. Furthermore, the mobility of oxygen ions is relatively lower in the dual-phase solid-electrolyte models than in the pure cubic models. This is due to their intrinsically unfavorable configuration for ion penetration. Comparison of the diffusion coefficient and ionic conductivity of the oxygen ions in various phase compositions shows an obvious effect of phase composition complexity in the systems with high Sc(2)O(3) concentration. The introduction of the dual-phase composition concept for the MD simulation of oxygen-ion conduction behavior is a promising approach to obtaining a more realistic understanding of ion transport in a solid electrolyte. PMID- 19562789 TI - Precisely programmed and robust 2D streptavidin nanoarrays by using periodical nanometer-scale wells embedded in DNA origami assembly. AB - A new punched DNA origami assembly with periodic nanometer-scale wells has been successfully designed and constructed. Through the attachment of two biotins at the two edges of each well, just one streptavidin (SA) tetramer (d = 5 nm) was size-selectively captured in each 6.8 x 12 x 2.0 nm well; this allowed formation of a 28 nm-period SA nanoarray of individual molecules. The position of SA capture can be fully controlled by placement of biotins in the nanoarray well. Moreover, construction of a 2D nanoarray of individual SA tetramers through selective positioning of SA tetramers in any desired wells in a complex of such punched origami motifs is also possible. The stability of the SA captured by this fixation strategy (DNA wells and two biotin linkers) was directly compared on the same molecule with the stability of SA captured with other possible strategies that do not employ wells or two linkers. In this way, the robustness of this means of fixation was clearly established. PMID- 19562792 TI - Jiggling of coherent excitons along a polymer chain. PMID- 19562793 TI - MS binding assays-with MALDI toward high throughput. AB - A novel type of MS binding assay, a substitute for radioligand binding, in which the quantification of the MS marker is performed by MALDI-MS-MS (FlashQuant) has been established. Because conventional MS binding assays can only be carried out by LC-ESI-MS-MS, the use of the FlashQuant system substantially increases the throughput capacity of this method. The study was performed for mGAT1 as a model system. First, a method was developed to quantify NO 711 as a marker for mGAT1 in a range from 208 pM to 16.7 nM using [2H10]NO 711 as an internal standard. On this basis, MS binding assays for mGAT1 could be implemented. Affinity constants determined in both saturation and competition experiments were in excellent agreement with those obtained in MS binding assays based on LC-ESI-MS-MS quantification. As the MALDI-MS system takes only a few seconds for quantification per sample, and the whole assay procedure is executed in a 96-well format, this technique is amenable to high-throughput screening. PMID- 19562794 TI - Discovery of 2-naphthoic acid monooxygenases by genome mining and their use as biocatalysts. AB - The large pool of cytochrome P450 (P450) open-reading frames identified in genome sequences has attracted much attention as a resource for new oxidation biocatalysts. P450 genes were cloned from genome-sequenced bacteria and coexpressed with putidaredoxin and its reductase genes to provide the redox partners of P450 in Escherichia coli. Whole-cell assays were performed with 2 naphthoic acid as a substrate. Hydroxylated naphthoic acid products were rapidly detected with two reagents showing different colors in the presence of the products. Two P450s, CYP199A1 and CYP199A2, were found to hydroxylate the substrate to 7- and 8-hydroxy-2-naphthoic acids. The CYP199A1 whole-cell biocatalyst converted 1 mM 2-naphthoic acid to 0.27 mM 7-hydroxy-2-naphthoic acid and 0.53 mM 8-hydroxy-2-naphthoic acid. CYP199A2 exhibited similar regioselectivity to CYP199A1. Furthermore, we found that 8-hydroxy-2-naphthoic acid emits near-white fluorescence when exposed to UV light. These P450s will provide a facile and environmentally friendly synthetic approach to the hydroxynaphthoic acids. PMID- 19562796 TI - A proteomics study of barley powdery mildew haustoria. AB - A number of fungal and oomycete plant pathogens of major economic importance feed on their hosts by means of haustoria, which they place inside living plant cells. The underlying mechanisms are poorly understood, partly due to difficulty in preparing haustoria. We have therefore developed a procedure for isolating haustoria from the barley powdery mildew fungus (Blumeria graminis f.sp. hordei, Bgh). We subsequently aimed to understand the molecular mechanisms of haustoria through a study of their proteome. Extracted proteins were digested using trypsin, separated by LC, and analysed by MS/MS. Searches of a custom Bgh EST sequence database and the NCBI-NR fungal protein database, using the MS/MS data, identified 204 haustoria proteins. The majority of the proteins appear to have roles in protein metabolic pathways and biological energy production. Surprisingly, pyruvate decarboxylase (PDC), involved in alcoholic fermentation and commonly abundant in fungi and plants, was absent in our Bgh proteome data set. A sequence encoding this enzyme was also absent in our EST sequence database. Significantly, BLAST searches of the recently available Bgh genome sequence data also failed to identify a sequence encoding this enzyme, strongly indicating that Bgh does not have a gene for PDC. PMID- 19562797 TI - Validating divergent ORF annotation of the Mycobacterium leprae genome through a full translation data set and peptide identification by tandem mass spectrometry. AB - Mycobacterium leprae has undergone extensive degenerative evolution, with a large number of pseudogenes. It is also the organism with the greatest divergence between gene annotations from independent institutes. Therefore, M. leprae is a good model to verify the currently predicted coding sequence regions between different annotations, to identify new ones and to investigate the expression of pseudogenes. We submitted a total extract of the bacteria isolated from Armadillo to Gel-LC-MS/MS using a linear quadrupole ion trap-Orbitrap mass spectrometer. Spectra were analyzed using the Leproma (1614 genes and 1133 pseudogenes) and TIGR (5446 genes) databases and a database containing the full genome translation. We identified a total of 1046 proteins, including five proteins encoded by previously predicted pseudogenes, which upon closer inspection appeared to be proper genes. Only 11 of the additional annotations by TIGR were verified. We also identified six tryptic peptides from five proteins from regions not considered to be coding sequences, in addition to peptides from two unannotated gene candidates that overlap with other genes. Our data show that the Leproma annotation of M. leprae is quite accurate, and there were no peptide observations corresponding to true pseudogenes, except for a new gene candidate, overlapping with an essential enolase on the complementary strand. PMID- 19562798 TI - Attenuation of inflammation and cellular stress-related pathways maintains insulin sensitivity in obese type I interleukin-1 receptor knockout mice on a high-fat diet. AB - The development of insulin resistance in the obese is associated with chronic, low-grade inflammation. We aimed to identify novel links between obesity, insulin resistance and the inflammatory response by comparing C57BL/6 with type I interleukin-1 receptor knockout (IL-1RI(-/-)) mice, which are protected against diet-induced insulin resistance. Mice were fed a high-fat diet for 16 wk. Insulin sensitivity was measured and proteomic analysis was performed on adipose, hepatic and skeletal muscle tissues. Despite an equal weight gain, IL-1RI(-/-) mice had lower plasma glucose, insulin and triacylglycerol concentrations, compared with controls, following dietary treatment. The higher insulin sensitivity in IL-1RI( /-) mice was associated with down-regulation of antioxidant proteins and proteasomes in adipose tissue and hepatic soluble epoxide hydrolase, consistent with a compromised inflammatory response as well as increased glycolysis and decreased fatty acid beta-oxidation in their muscle. Their lower hepatic triacylglycerol concentrations may reflect decreased flux of free fatty acids to the liver, decreased hepatic fatty acid-binding protein expression and decreased lipogenesis. Correlation analysis revealed down-regulation of classical biomarkers of ER stress in their adipose tissue, suggesting that disruption of the IL-1RI-mediated inflammatory response may attenuate cellular stress, which was associated with significant protection from diet-induced insulin resistance, independent of obesity. PMID- 19562799 TI - Comprehensive proteomic and transcriptomic analysis reveals early induction of a protective anti-oxidative stress response by low-dose proteasome inhibition. AB - Effective inhibition of the proteasome by high doses of proteasome inhibitors induces apoptotic cell death. In contrast, partial proteasome inhibition by low inhibitor doses mediates a protective cellular stress response. The early targets and mediators of these dose-dependent effects of proteasome inhibitors are unknown. Primary human umbilical cord vein endothelial cells were treated with low and high doses of the proteasome inhibitor MG132 for 2 h. In a combined 2-DE and MS approach, we identified more than 20 new targets of proteasome inhibition. These proteins are involved in cell cycle regulation, signaling, cytoskeletal rearrangement, and cellular stress response. Accompanying Affymetrix analysis revealed that these proteins are not regulated on the transcriptional level but are mainly stabilized by proteasome inhibition. The proteasome-dependent accumulation of the anti-oxidative sensor proteins DJ-1, peroxiredoxin-1 and -6 was accompanied by dose-dependent induction of oxidative stress after 2 h of proteasome inhibition and contributed to the differential transcriptional stress response to low- and high-dose proteasome inhibition: Whereas low-dose proteasome inhibition induces a transcriptional profile reminiscent of a physiological stress response that preconditions and protects endothelial cells from oxidative stress, high inhibitor doses induce massive transcriptional dysregulation and pronounced oxidative stress triggering apoptosis. PMID- 19562800 TI - Protein composition of oil bodies from mature Brassica napus seeds. AB - Seed oil bodies (OBs) are intracellular particles storing lipids as food or biofuel reserves in oleaginous plants. Since Brassica napus OBs could be easily contaminated with protein bodies and/or myrosin cells, they must be purified step by step using floatation technique in order to remove non-specifically trapped proteins. An exhaustive description of the protein composition of rapeseed OBs from two double-zero varieties was achieved by a combination of proteomic and genomic tools. Genomic analysis led to the identification of sequences coding for major seed oil body proteins, including 19 oleosins, 5 steroleosins and 9 caleosins. Most of these proteins were also identified through proteomic analysis and displayed a high level of sequence conservation with their Arabidopsis thaliana counterparts. Two rapeseed oleosin orthologs appeared acetylated on their N-terminal alanine residue and both caleosins and steroleosins displayed a low level of phosphorylation. PMID- 19562801 TI - Proteomics analysis of two mice hepatocarcinoma ascites syngeneic cell lines with high and low lymph node metastasis rates provide potential protein markers for tumor malignancy attributes to lymphatic metastasis. AB - Lymph node metastasis (LNM) is recognized as an important factor involved in the tumor malignancy progression. Our previous study has indicated that the hepatocarcinoma cell line with 75% of LNM (Hca-F)-cell-induced neoplasia and the hepatocarcinoma cell line with 25% of LNM-induced neoplasia are accompanied with high (75%) and low (25%) incidences of LNM. In the current study, 62 and 54 protein spots were observed up-regulated and down-regulated in Hca-F cell relative to the hepatocarcinoma cell line with 25% of LNM by 2-D DIGE. Totally, 113 unique proteins were identified by HPLC-nano ESI-MS/MS analysis. The expression levels of Annexin A7, Ulch3, and ER protein 29 were validated by Western blotting analyses. The abnormally regulated proteins were categorized and annotated by protein analysis through evolutionary relationships analysis with the aid of the database for annotation, visualization and integrated discovery tool. Seventeen gene candidates concordantly expressed both at mRNA and protein levels. By making a challenge, we detected expression levels of Annexin A7 in primary gastric cancer (GC) and primary GC cancer tissues with LNMs by immunohistochemisty. Higher ratio of positive and strong expressions Annexin A7 in GC might correlate with the tumor progression. The repression of Annexin A7 inhibits the mobility and invasion abilities of Hca-F cell, increases the apoptosis rate of Hca-F cell. Current study narrows and provides certain specific protein candidates potentially playing important roles in LNM-associated cancers. PMID- 19562803 TI - 2-D DIGE analysis implicates cytoskeletal abnormalities in psychiatric disease. AB - The mechanisms underlying white matter changes in psychiatric disease are not known. We aimed to characterise the differential protein expression in deep white matter from the dorsolateral prefrontal cortex from 35 schizophrenia, 35 bipolar disorder, and 35 control subjects, from the Stanley Array Collection. We used 2-D DIGE to profile for protein expression changes in the brain. We found 70 protein spots to be significantly differentially expressed between disease and control subjects (ANCOVA, p<0.05), 46 of which were subsequently identified by LC-MS/MS. The proteins identified included novel disease candidates as well as proteins that have previously been reported as abnormal in schizophrenia, thus reinforcing their association with the disease. Furthermore, we confirmed the direction of change for three proteins using ELISA, namely neurofilament-light, amphiphysin II, and Rab-GDP-alpha, in a subset of the Stanley Array Collection. In addition, altered expression of neurofilament-light, amphiphysin II, and Rab-GDP-alpha was not observed in the cortex of mice chronically treated with haloperidol, making it less likely that these alterations are a consequence of neuroleptic medication. The data presented here strongly suggest disruption of the cytoskeleton and its associated signal transduction proteins in schizophrenia, and to a lesser extent in bipolar disorder. PMID- 19562804 TI - Effect of phosphorylated hsp27 on proliferation of human endothelial and smooth muscle cells. AB - Recent studies have suggested a protective role of hsp27 against atherosclerosis and transplant graft vasculopathy. Here we have investigated the effects of over expression of wild-type hsp27 and its phosphorylation mimics on proliferation of human endothelial cells (ECs) and smooth muscle cells (SMCs). ECs and SMCs cultured from human blood vessels or cells lines (human microvascular endothelial cell line and human telomerase reverse transcriptase subunit SMC) were infected with adenovirus containing DNA from wild-type hsp27, hyper-phosphorylated hsp27 mimic (3D hsp27), hypo-phosphorylated hsp27 mimic (3A hsp27) or anti-sense hsp27, and proliferation measured over the next 5 days. Protein extracts from infected cells were subjected to proteomic analysis using 2-D DIGE. Over-expression of 3D hsp27 and anti-sense hsp27 but not 3A hsp27 mimic caused significant inhibition of proliferation of ECs and SMCs. Proteomic analysis focussed on proteins that were significantly down-regulated by the 3D hsp27 mutant. The cell cycling proteins stathmin, cofilin and ubiquitination enzymes fullfilled these criteria. 1-D Western blots of infected human microvascular endothelial cell line and human telomerase reverse transcriptase subunit SMC confirmed down-regulation of stathmin, cofilin and ubiquitination enzymes by 3D hsp27. The phosphorylation status of hsp27 is an important regulator of proliferation of human vascular ECs and SMCs; possibly contributing to cardiovascular protection. PMID- 19562802 TI - Systems approach to explore components and interactions in the presynapse. AB - The application of proteomic techniques to neuroscientific research provides an opportunity for a greater understanding of nervous system structure and function. As increasing amounts of neuroproteomic data become available, it is necessary to formulate methods to integrate these data in a meaningful way to obtain a more comprehensive picture of neuronal subcompartments. Furthermore, computational methods can be used to make biologically relevant predictions from large proteomic data sets. Here, we applied an integrated proteomics and systems biology approach to characterize the presynaptic (PRE) nerve terminal. For this, we carried out proteomic analyses of presynaptically enriched fractions, and generated a PRE literature-based protein-protein interaction network. We combined these with other proteomic analyses to generate a core list of 117 PRE proteins, and used graph theory-inspired algorithms to predict 92 additional components and a PRE complex containing 17 proteins. Some of these predictions were validated experimentally, indicating that the computational analyses can identify novel proteins and complexes in a subcellular compartment. We conclude that the combination of techniques (proteomics, data integration, and computational analyses) used in this study are useful in obtaining a comprehensive understanding of functional components, especially low-abundance entities and/or interactions in the PRE nerve terminal. PMID- 19562805 TI - Quantitative analysis of phosphopeptides in search of the disease biomarker from the hepatocellular carcinoma specimen. AB - Reversible phosphorylation of proteins is the most common PTM in cell-signaling pathways. Despite this, high-throughput methods for the systematic detection, identification, and quantification of phosphorylated peptides have yet to be developed. In this paper, we describe the establishment of an efficient online titaniuim dioxide (TiO2)-based 3-D LC (strong cationic exchange/TiO2/C18)-MS(3) linear ion trap system, which provides fully automatic and highly efficient identification of phosphorylation sites in complex peptide mixtures. Using this system, low-abundance phosphopeptides were isolated from cell lines, plasma, and tissue of healthy and hepatocellular carcinoma (HCC) patients. Furthermore, the phosphorylation sites were identified and the differences in phosphorylation levels between healthy and HCC patient specimens were quantified by labeling the phosphopeptides with isotopic analogs of amino acids (stable isotope labeling with amino acids in cell culture for HepG2 cells) or water (H(2) (18)O for tissues and plasma). Two examples of potential HCC phospho-biomarkers including plectin-1(phopho-Ser-4253) and alpha-HS-glycoprotein (phospho-Ser 138 and 312) were identified by this analysis. Our results suggest that this comprehensive TiO2-based online-3-D LC-MS(3)-linear ion trap system with high-throughput potential will be useful for the global profiling and quantification of the phosphoproteome and the identification of disease biomarkers. PMID- 19562806 TI - Giant cyclo[n]thiophenes with extended pi conjugation. PMID- 19562808 TI - Gold catalysis: switching the pathway of the furan-yne cyclization. PMID- 19562809 TI - A single alpha-helical turn stabilized by replacement of an internal hydrogen bond with a covalent ethylene bridge. PMID- 19562811 TI - Stereospecific C-H oxidation with H2O2 catalyzed by a chemically robust site isolated iron catalyst. PMID- 19562807 TI - Nanogels as pharmaceutical carriers: finite networks of infinite capabilities. AB - Nanogels are swollen nanosized networks composed of hydrophilic or amphiphilic polymer chains. They are developed as carriers for the transport of drugs, and can be designed to spontaneously incorporate biologically active molecules through formation of salt bonds, hydrogen bonds, or hydrophobic interactions. Polyelectrolyte nanogels can readily incorporate oppositely charged low-molecular mass drugs and biomacromolecules such as oligo- and polynucleotides (siRNA, DNA) as well as proteins. The guest molecules interact electrostatically with the ionic polymer chains of the gel and become bound within the finite nanogel. Multiple chemical functionalities can be employed in the nanogels to introduce imaging labels and to allow targeted drug delivery. The latter can be achieved, for example, with degradable or cleavable cross-links. Recent studies suggest that nanogels have a very promising future in biomedical applications. PMID- 19562812 TI - HIV: a discovery opening the road to novel scientific knowledge and global health improvement (Nobel lecture). PMID- 19562810 TI - Long-range correlations between aliphatic 13C nuclei in protein MAS NMR spectroscopy. PMID- 19562813 TI - Bis(allyl)calcium. PMID- 19562814 TI - The physical origin of Saytzeff's rule. PMID- 19562815 TI - Hydrogen-bond control in axially chiral styrenes: selective synthesis of enantiomerically pure C2-symmetric paracyclophanes. PMID- 19562816 TI - The true structures of the vannusals, part 1: Initial forays into suspected structures and intelligence gathering. PMID- 19562817 TI - The true structures of the vannusals, part 2: Total synthesis and revised structure of vannusal B. PMID- 19562818 TI - Long-range, high-resolution magnetic imaging of nanoparticles. PMID- 19562819 TI - Dendritic DNA building blocks for amplified detection assays and biomaterials. PMID- 19562820 TI - Exchange-coupled bimagnetic wustite/metal ferrite core/shell nanocrystals: size, shape, and compositional control. PMID- 19562821 TI - Mechanistic surprises in the gold(I)-catalyzed intramolecular hydroarylation of allenes. PMID- 19562822 TI - A self-assembled protein nanotube with high aspect ratio. AB - Production of a self-assembled protein nanotube achieved through engineering of the 11mer ring protein trp RNA-binding attenuation protein is described. The produced mutant protein is able to stack in solution to produce an extremely narrow, uniform nanotube apparently stabilized by a mixture of disulfide bonds and hydrophobic interactions. Assembly is reversible and the length of tube can potentially be controlled. Large quantities of hollow tubes 8.5 nm in overall diameter with lengths varying from 7 nm to over 1 microm are produced. The structure is analyzed using transmission electron microscopy, atomic force microscopy, mass spectrometry, and single-particle analysis and it is found that component rings stack in a head-to-head fashion. The internal diameter of the tube is 2.5 nm, and the amino acid residues lining the central cavity can be mutated, raising the possibility that the tube can be filled with a variety of conducting or semiconducting materials. PMID- 19562823 TI - Cysteine-capped ZnSe quantum dots as affinity and accelerating probes for microwave enzymatic digestion of proteins via direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis. AB - Fluorescent semiconductor quantum dots (QDs) exhibit great potential and capability for many biological and biochemical applications. We report a simple strategy for the synthesis of aqueous stable ZnSe QDs by using cysteine as the capping agent (ZnSe-Cys QDs). The ZnSe QDs can act as affinity probes to enrich peptides and proteins via direct matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOFMS) analysis. This nanoprobe could significantly enhance protein signals (insulin, ubiquitin, cytochrome c, myoglobin and lysozyme) in MALDI-TOFMS by 2.5-12 times compared with the traditional method. Additionally, the ZnSe-Cys QDs can be applied as heat absorbers (as accelerating probes) to speed up microwave-assisted enzymatic digestion reactions and also as affinity probes to enrich lysozyme-digested products in MALDI-TOFMS. Furthermore, after the enrichment experiments, the solutions of ZnSe-Cys QDs mixed with proteins can be directly deposited onto the MALDI plates for rapid analysis. This approach shows a simple, rapid, efficient and straightforward method for direct analysis of proteins or peptides by MALDI TOFMS without the requirement for further time-consuming separation processes, tedious washing steps or laborious purification procedures. The present study has demonstrated that ZnSe-Cys QDs are reliable and potential materials for rapid, selective separation and enrichment of proteins as well as accelerating probes for microwave-digested reactions for proteins than the regular MALDI-MS tools. Additionally, we also believe that this work may also inspire investigations for applications of QDs in the field of MALDI-MS for proteomics. PMID- 19562824 TI - Virulence development and genetic polymorphism in Meloidogyne incognita (Kofoid & White) Chitwood after prolonged exposure to sublethal concentrations of nematicides and continuous growing of resistant tomato cultivars. AB - BACKGROUND: The root-knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood, is an important plant pathogen damaging to tomato. Continuous use of resistant tomato cultivars and nematicides for its effective management might lead to resistance break-up or nematicide failure. Genetic variability and virulence in M. incognita on susceptible Pusa Ruby tomato were analysed by bioassay, esterase and DNA polymorphism after a 5 year weekly exposure to carbofuran, carbosulfan, cadusafos and triazophos at 0.0125, 0.0250 and 0.0500 microg g(-1). Virulence in M. incognita after a 5 year multiplication on resistant tomatoes was assessed. RESULTS: The nematicidal treatments resulted in the development of virulent M. incognita populations. Their invasion potential increased significantly after continuous exposure to low concentrations of the nematicides. Also, growing resistant tomato cultivars for ten successive seasons resulted in a 6.6% increase in the invasion potential. These virulent populations exhibited 1-3 additional esterase and DNA bands compared with untreated populations. CONCLUSION: A 5 year exposure of M. incognita to sublethal concentrations of nematicides or resistant tomato cultivars exerted enough selection pressure to cause genomic alterations for virulence development. Isozyme markers can be used for rapid and precise diagnostics of field populations by advisory services, enabling judicious remedial management decisions. PMID- 19562825 TI - Menarche in girls and headache--a longitudinal analysis. AB - BACKGROUND: Puberty is assumed to influence the occurrence of headache, increasing the risk for recurrent episodes, especially in girls. The increase of headache, in particular recurrent headache, in girls from around the age of 12 on, is often ascribed to the occurrence of menarche as the most stringent indicator of completed puberty. OBJECTIVES: We examined the hypothesis that the occurrence of menarche in girls is predictive of recurrent headache in reference to no or rare headache in the past 6 months. Furthermore, the assumption was tested that headache episodes increase after onset of menarche but remain unchanged in girls not having experienced their first menstruation. We also expected a higher probability of migraine type of headache after menarche. In a further analysis girls with and without menarche were compared with boys, and a difference between gender only for girls with menarche was predicted. METHODS: In an epidemiological study recruiting 8800 families with children 7-14 years in Southern Lower Saxony (Germany), we conducted 3 yearly panels based on postal questionnaires. Headache data are based on self-report of children from 9 years on. Parents were asked for information regarding the onset of menarche. Approximately 1100 girls and about the same number of boys constitute the sample for our analyses. RESULTS: The results of a logistic regression analysis demonstrate that in 11- to 16-year-old girls having experienced menarche in either the year of the assessment of headache or 2 years before that, the risk for recurrent headache is increased when compared with girls without menarche. Intraindividual longitudinal analyses, however, do not support our hypotheses that after the onset of menarche headaches become more frequent. The expected differences in headache between girls with menarche and boys were found. The risk for migraine-like headaches is not significantly higher after onset of menarche than before. CONCLUSIONS: Thus, results do not consistently support the hypothesis that puberty is a moderator of headache frequency. Even in cases where the outcome of regression analyses is supportive of our expectations, explained variance is diminutive (maximum 2.2%). Thus, the influence of menarche on headache seems to be only marginal. PMID- 19562826 TI - Incomplete posterior circle of willis: a risk factor for migraine? AB - BACKGROUND: Migraine is associated with vascular risk factors and white matter abnormalities (WMA). Cerebral hypoperfusion is known to be one mechanism underlying WMA and a few studies have shown that an incomplete circle of Willis (CW) may predispose to cerebral hypoperfusion. This study assessed the relationship between the morphologic characteristics of the CW and migraine. METHODS: This case-control study was carried out in the Amiens University Hospital. Patients undergoing 3-dimensional time of flight magnetic resonance angiography of the CW from January 1 to June 30, 2006 were included (n = 124). A definitive diagnosis of migraine was established in 47 patients: 23 (48.9%) experienced migraine without aura and 24 (51.1%) migraine with aura. The remaining 77 patients with other neurologic disorders constituted the control group. The posterior CW was graded as complete when both posterior communicating arteries and the P1 segments of the posterior cerebral artery were present on visual examination and incomplete when one of these vessels was missing (interobserver agreement: K(total) = 0.746). RESULTS: Incomplete posterior CW was significantly more common in migraineurs than in the control group (49% vs 18%; P < .001). On multivariate analysis, incomplete posterior CW was the sole independent factor associated with migraine (OR: 6.5; 95% CI: 2.6-16.2; P < .001). No difference was found between migraineurs with and without aura. CONCLUSIONS: Despite some methodological limitations, our results showed that incomplete posterior CW was associated with migraine. PMID- 19562827 TI - SUNA syndrome with seasonal pattern. AB - SUNA is a trigeminal autonomic cephalalgia (TAC) characterized by short unilateral attacks centered on the ophthalmic trigeminal distribution, and accompanied by at least one of a number of cranial autonomic symptoms that can include lacrimation, redness of the ipsilateral eye, nasal congestion, rhinorrhea, and eyelid edema. It exists in episodic and chronic form. We have described an atypical case of episodic SUNA with an exclusive seasonal pattern as previously reported in other trigeminal autonomic cephalalgia, commonly known as TACs. PMID- 19562828 TI - Isolated cortical venous thrombosis associated with intracranial hypotension syndrome. AB - The association of intracranial hypotension syndrome with cerebral venous thrombosis is rare. We report our experience with isolated cortical venous thrombosis, which developed after unsuccessful epidural anesthesia. Magnetic resonance imaging showed characteristic imaging findings of intracranial hypotension syndrome, such as dural thickening and brain sagging. We also detected right parietal venous hemorrhagic infarction secondary to right-sided cortical venous thrombosis. After the treatment of intracranial hypotension via epidural blood patch, heparin was used to treat cortical venous thrombosis. PMID- 19562829 TI - Dentigerous cyst presenting with sinus headache as the primary symptom. PMID- 19562830 TI - Ibn Sina (Avicenna) on pathogenesis of migraine compared with the recent theories. PMID- 19562832 TI - Signalling mechanisms in mouse bitter responsive taste cells. AB - To clarify the components involved in mouse intracellular bitter transduction in the mouse, we investigated the expression patterns of transduction components and the contribution of intracellular Ca2+ stores. Of 64 cells responding to bitter compounds, 63% of the cells expressed the taste tissue-specific G-protein alpha gustducin. Of the alpha-gustducin immunoreactive cells, 78% expressed IP3R3, a receptor that mediates Ca2+ release from the intracellular store. When intracellular Ca2+ was depleted by treatment with the inhibitor thapsigargin, 83% of the taste cells lost their response to bitter. These data suggest that bitterness transduction involves the release of Ca2+ from intracellular stores. PMID- 19562831 TI - Sex differences in brain structure in auditory and cingulate regions. AB - We applied a new method to visualize the three-dimensional profile of sex differences in brain structure based on MRI scans of 100 young adults. We compared 50 men with 50 women, matched for age and other relevant demographics. As predicted, left hemisphere auditory and language-related regions were proportionally expanded in women versus men, suggesting a possible structural basis for the widely replicated sex differences in language processing. In men, primary visual, and visuo-spatial association areas of the parietal lobes were proportionally expanded, in line with prior reports of relative strengths in visuo-spatial processing in men. We relate these three-dimensional patterns to prior functional and structural studies, and to theoretical predictions based on nonlinear scaling of brain morphometry. PMID- 19562833 TI - Controlled delivery of bovine serum albumin from carboxymethyl xanthan microparticles. AB - Bovine serum albumin (BSA)-loaded carboxymethyl xanthan (CMX) microparticles were prepared following gelation of sodium carboxymethyl xanthan (SCMX) gum with different concentrations (1-5%) of aluminium chloride (AlCl3). The microparticles prepared using 1% AlCl3 were subsequently coated with 0.5% aqueous solution of either SCMX gum or sodium alginate. Both uncoated and coated microparticles were characterized for entrapment efficiency, surface morphology, particle size, in vitro release and protein stability. The uncoated microparticles became non spherical and the mean diameter was found to increase with increasing AlCl3 concentration. Higher concentration of AlCl3 decreased BSA entrapment efficiency of the uncoated microparticles from 86-61%. Furthermore, BSA entrapment in coated microparticles was found lower (78-79%) than uncoated microparticles prepared using 1% AlCl3. Although, the uncoated microparticles released almost half of its content in NaCl-HCl buffer solution (pH 1.2) in 2 h, the alginate and xanthan coated microparticles did not liberate a substantial amount of entrapped protein within the same period and prolonged the release in PBS solution (pH 7.4) up to 10 and 12 h, respectively. The microparticles released the protein via diffusion and swelling of the polymer matrix. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that BSA integrity was well retained in the CMX microparticles. PMID- 19562834 TI - Evolution of prenatal diagnostic techniques from phenotypic diagnosis to gene arrays: its likely impact on prenatal diagnosis of hemophilia. AB - Prenatal diagnostic techniques in hemophilia have evolved through the early sex determination techniques of offering a nonspecific diagnosis in case of a male fetus through the various mutation screening techniques to the more recent gene array techniques. Each of these techniques has specific advantages and disadvantages. The sampling techniques have evolved simultaneously to suit the requirements of each technique and also the different gestation periods. The DNA based testing methods provide a range of aberrations detected with different levels of genomic resolution. The more recent gene array analysis is poised to have substantial impact on prenatal diagnosis of hemophilia not only in studying the highly heterogeneous mutations but may also be useful in studying the effect of various ameliorating or epistatic genetic mutations/ polymorphisms simultaneously, providing a wide range of options to the prenatal diagnosis experts, the genetic counselors, and the couples opting for prenatal diagnosis. PMID- 19562836 TI - [After-hours-care and general practice]. PMID- 19562835 TI - Acute myocardial infarction due to oral contraceptive. PMID- 19562837 TI - Clinical and research work in people with prodromal signs of psychosis. PMID- 19562838 TI - An interview with Jeffrey Sturchio. PMID- 19562839 TI - [Prognostic and predictive factors in different malignant diseases]. PMID- 19562840 TI - [Analysis of the metabolome suggests a major potential role of sarcosine in the progression of cancer of the prostate]. PMID- 19562841 TI - The graying of HIV. PMID- 19562842 TI - [Targetted combined radiotherapy and chemotherapy]. PMID- 19562843 TI - Mental health matters. PMID- 19562844 TI - Findings of research misconduct. PMID- 19562845 TI - Patients expect transparency in doctors' relationships with the pharmaceutical industry. Comment. PMID- 19562847 TI - Serum eosinophil-derived neurotoxin in childhood atopic dermatitis: a useful marker of disease activity? PMID- 19562848 TI - How could ORs benefit from the government's health IT funds? PMID- 19562846 TI - World's first implant of a 23-mm Carpentier-Edwards Perimount Magna mitral bioprosthesis. PMID- 19562849 TI - Involvement of Fas/FasL system in apoptotic signaling in testicular germ cells of male Wistar rats injected i.v. with microcystins. AB - Previous studies have shown that gonads were the second target organ of microcystins (MCs), and that MCs exposure exerted obvious toxic effects on male reproductive system of mammals. However, relevant molecular evidences are still lacking. Fas-signaling pathway plays a key role in toxicant-induced germ cell apoptosis. This study was to evaluate the responses of Fas/FasL system related genes and proteins in testes of rats injected intravenously with MCs. Enhanced apoptosis of germ cells in the testes of MCs-treated rats was detected by the terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick end labeling (TUNEL) associated with up-regulation of the Fas/FasL system. Both Fas and FasL protein expression were induced evidently from 1 h post-injection, and this high expression level maintained throughout the experiment. In addition, the activation of caspase-8 and caspase-3 protein was also observed, which were indicators of apoptosis. These results suggested the likely involvement of Fas/FasL system in the MCs-induced germ cell apoptosis. It is also suggested that MCs can cause damage to Sertoli cells directly. PMID- 19562850 TI - [Multidisciplinary management of severe pre-eclampsia]. PMID- 19562851 TI - By the way, doctor. Is it safe for children to drink soy milk? PMID- 19562852 TI - By the way, doctor. I've been having PET-CT scans to look for any growth or spread of previously removed tumors. Are they as reliable in detecting breast cancer as a regular mammogram? PMID- 19562853 TI - By the way, doctor. I've had type 2 diabetes for 12 years; I'm now 81. I take metformin plus Januvia. My doctor has never suggested that I monitor myself on a daily basis with a meter. My hemoglobin A1c has been creeping up and is now at 7. Should I be getting more aggressive in my treatment? PMID- 19562854 TI - By the way, doctor. I read in the April issue that one of the nutrition experts eats five servings of fish a week. Why so much? And isn't there a risk from the contaminants? PMID- 19562855 TI - By the way, doctor. I have been told by a podiatrist that because of hammertoes, the fat pods on the ball of my foot have dissipated, causing pain when I walk or stand for long periods. I like to walk for exercise so this is a real bother. I am trying to figure out how to handle the foot and all of its idiosyncrasies. PMID- 19562856 TI - By the way, doctor. I have a plugged left ear problem. It usually starts about 4 p.m. and lasts about three to four hours. I cough, clear my throat, yawn, close my nostrils and blow--nothing seems to help. PMID- 19562857 TI - By the way, doctor. I am curious about tremors in adults. I know that people with Parkinson's disease get them, but I am not sure about other conditions. PMID- 19562858 TI - By the way, doctor. About 10 years ago, you published an article saying that Ginkgo biloba might slow down dementia. I was 59, and I started taking it. I think it has helped me and know others who think likewise. But I read that a recent study says it doesn't help. Your thoughts? PMID- 19562859 TI - Patients expect transparency in doctors' relationships with the pharmaceutical industry. Comment. PMID- 19562860 TI - Swine-origin influenza A H1N1 update. PMID- 19562861 TI - Investment should follow Obama's pledges on HIV/AIDS. PMID- 19562862 TI - [Quality of life in Italian provinces]. PMID- 19562863 TI - The cardiac literature 2008. PMID- 19562865 TI - Ethical issues and innovations in colorectal surgery. PMID- 19562864 TI - Position of the American Dietetic Association: vegetarian diets. AB - It is the position of the American Dietetic Association that appropriately planned vegetarian diets, including total vegetarian or vegan diets, are healthful, nutritionally adequate, and may provide health benefits in the prevention and treatment of certain diseases. Well-planned vegetarian diets are appropriate for individuals during all stages of the life cycle, including pregnancy, lactation, infancy, childhood, and adolescence, and for athletes. A vegetarian diet is defined as one that does not include meat (including fowl) or seafood, or products containing those foods. This article reviews the current data related to key nutrients for vegetarians including protein, n-3 fatty acids, iron, zinc, iodine, calcium, and vitamins D and B-12. A vegetarian diet can meet current recommendations for all of these nutrients. In some cases, supplements or fortified foods can provide useful amounts of important nutrients. An evidence- based review showed that vegetarian diets can be nutritionally adequate in pregnancy and result in positive maternal and infant health outcomes. The results of an evidence-based review showed that a vegetarian diet is associated with a lower risk of death from ischemic heart disease. Vegetarians also appear to have lower low-density lipoprotein cholesterol levels, lower blood pressure, and lower rates of hypertension and type 2 diabetes than nonvegetarians. Furthermore, vegetarians tend to have a lower body mass index and lower overall cancer rates. Features of a vegetarian diet that may reduce risk of chronic disease include lower intakes of saturated fat and cholesterol and higher intakes of fruits, vegetables, whole grains, nuts, soy products, fiber, and phytochemicals. The variability of dietary practices among vegetarians makes individual assessment of dietary adequacy essential. In addition to assessing dietary adequacy, food and nutrition professionals can also play key roles in educating vegetarians about sources of specific nutrients, food purchase and preparation, and dietary modifications to meet their needs. PMID- 19562866 TI - Tumor necrosis factor inhibitors in the treatment of chronic inflammatory diseases. A review of immunogenicity and potential implications. PMID- 19562867 TI - [The use of recombinant activated factor VII in traumatic intracranial haemorrhage]. AB - INTRODUCTION: A group of patients with craniocerebral injuries received recombinant activated factor VII (rFVIIa) in order to reduce or stop further bleeding in brain tissue. OBJECTIVE: Fundamental objective was to show that the use of rFVIIa can stop intracerebral bleeding and that operative treatment of such patients can be avoided. METHODS: rFVIIa was applied in the group of patients who sustained traumatic brain injuries. The control group was formed retrospectively out of patients with the same pathology previously treated at this clinic throughout 2003 and 2004. It is important to mention that in all patients rFVIIa was applied within first four hours after injury. Coagulation status and GCS were monitored in all patients. RESULTS: The treated patients had different mechanism of injury and pathological substrate of brain tissue. Results were compared with the group of patients who had similar pathological substrate and GCS on admission but did not receive rFVIIa. There were no complications or lethal outcomes recorded in the group of the treated patients. CONCLUSION: Cessation of intracerebral bleeding after timely use of rFVIIa enabled the use of only pharmacological treatment and avoiding operative treatment in some patients. Results were encouraging. PMID- 19562868 TI - [Recombinant activated factor VII in the treatment of intractable non-surgical bleeding following major vascular procedures]. AB - INTRODUCTION: Recombinant form of activated factor VII (rFVIIa) has been approved for use in haemophiliacs with antibodies to factor VIII or factor IX. Recent studies and clinical experiences have showed that rFVIIa gives extreme haemostatic effects in patients with severe "nonhaemophilic" bleeding produced after trauma and major surgery. OBJECTIVE: We present our preliminary experience of the use of rFVIIa in vascular surgery where conventional haemostatic measures were inadequate. METHOD: There were 17 patients divided into three groups: Group I--6 patients with ruptured abdominal aortic aneurysms; Group II--7 patients with thoracoabdominal aortic aneurysms and 1 patient with acute complicated dissection of thoracic aorta type III; Group III--3 patients with retroperitoneal tumours involving great abdominal vessels. RESULTS: Clinical improvement was reported following the treatment in 14 of 17 patients in our study. Bleeding was successfully controlled as evidenced by improved haemodynamic parameters and decreased inotrope and transfusion requirement. CONCLUSION: More liberal use of rFVIIa in vascular patients is limited, because there is no randomized controlled trial proving efficacy and safety in vascular patients. We recommend the use of rFVIIa in vascular surgery only during and after operative treatment of thoracoabdominal aortic aneurysms, ruptured abdominal aortic aneurysms and retroperitoneal tumours involving aorta and/or inferior vena cava complicated with "non-surgical" massive uncontrolled bleeding. PMID- 19562869 TI - [Our experince with the use of recombinant activated factor VII in postpartum haemorrhage]. AB - INTRODUCTION: Massive obstetric bleeding is the most common cause of maternal mortality and morbidity. The first step in treatment of these patients is establishing the adequate circulatory volume. The primary goal of therapy is to identify and remove the cause of bleeding, with appropriate symptomatic and substitution therapy. Human recombinant activated factor VII (rFVIIa) is officially registered for the treatment of patients suffering from haemophilia with inhibitors. Its use has also proved successful in other congenital and acquired coagulopathies and in patients with acute non-haemophilic bleeding. A special significance is given to the application of rFVIIa in cases of obstetric haemorrhage, in order to avoid postpartum hysterectomy and occurrence of complications of haemorrhagic shock in obstetrics. OBJECTIVE: The aim of this study is to show our experience and results of the use of rFVIIa in the treatment of patients with massive postpartum bleeding. METHOD: The retrospective study encompassed six patients with primary postpartum haemorrhage treated with rFVIIa at our institution in the period from 2005 to 2007. RESULTS: The treated patients were divided into two groups. In the first group, there were three patients who underwent hysterectomy and who received rFVIIa over 24 hours after delivery. The second group consisted of three patients who received rFVIIa in the first 24 hours after delivery, before we decided to perform hysterectomy. The application of rFVIIa led to successful cessation of bleeding in all patients. Relevant side effects were not registered. CONCLUSION: The administration of rFVIIa in obstetrics should be considered for each patient before decision to apply hysterectomy, and it should certainly be applied in patients who want to preserve the uterus and fertile capability. According to our experience, in cases of postpartum hemorrhagia rFVIIa is to be administered in intravenous bolus doses of at least 90 mcg/kg, at least 6 hours after the onset of bleeding. rFVIIa is not an alternative to adequate surgical haemostasis; therefore, it needs to be administered after its detailed revision. PMID- 19562870 TI - [Recombinant activated factor VII and intraoperative use of cell saver in neurosurgical treatment of arteriovenous malformation]. AB - INTRODUCTION: Arteriovenous (AV) malformation of brain causes abundant intraoperative bleedings, because of increased blood flow, which complicates operative treatment. The use of cell saver for intraoperative salvage of blood and recombinant activated factor VII (rFVIIa) significantly reduces complications during operative treatment. CASE OUTLINE: We present a 29-year old male patient with AV malformation of the 4th degree on Spetzler-Martin scale. Because of the possibility of abundant bleeding, a cell saver (Sequestra 1000 Metronic, USA) was used, and to achieve and control adequate haemostasis, we used rFVIIa. CONCLUSION: The use of cell saver for intraoperative blood salvage and rFVIIa proved successful in operative treatment of AV malformation of 4th degree. PMID- 19562871 TI - [Acquired haemophilia A presenting in postpartum period--case report]. AB - INTRODUCTION: Acquired haemophilia A is a rare autoimmune disorder caused by factor VIII (FVIII) inhibitors. Patients may present with catastrophic bleeding, despite having no prior history of bleeding disorders. Acquired haemophilia A is a rare complication of pregnancy, typically appearing in the postpartum period. The patients usually present with bleeding related to vaginal delivery or Caesarean section. Management includes control of haemorrhage and eradication of the FVIII inhibitor. CASE OUTLINE: Acquired haemophilia A was diagnosed in our patient after profuse vaginal bleeding related to vaginal delivery (FVIII level 2%; FVIII inhibitor titar 16 BJ). Red cell, fresh frozen plasma and cryoprecipitate transfusions, as well as vaginal packing were ineffective. The administration of recombinant activated factor VII (rFVIIa) (NovoSeven) proved effective in stopping the bleeding. To facilitate eradication of the inhibitor prednisone (30 mg orally/d) was added. After 3 weeks of treatment, the inhibitor FVIII titar was reduced to 50%. CONCLUSION: Acquired postpartum haemophilia must be always considered in the differential diagnosis of postpartum haemorrhage. According to our experience and reported literature (case reports and small series), initial haemodynamic stabilization after rFVIIa (NovoSeven) administration followed by immunosuppressive therapy is highly successful. Thus, most women presenting with acguired hemophilia A in the portpartum period has favourable prognosis. PMID- 19562872 TI - [Application of recombinant activated factor VII in treatment of intracranial haemorrhage in haemophilic patient with inhibitor]. AB - INTRODUCTION: Intracranial haemorrhage (ICH) is one of serious haemorrhagic syndromes among haemophilic patients and the potential cause of death. However, extensive use of coagulation factor concentrates can provoke the development of antibodies, i.e. inhibitors. Recombinant activated factor VII (rFVIIa) has been successfully used in treating haemorrhage in A or B haemophilic patients with inhibitors. CASE OUTLINE: A 31-year old patient was hospitalized at the local Clinical Centre to be prepared for dental surgery. His initial level of inhibitor was two Bethesda units. His condition deteriorated abruptly during hospitalization with signs of drowsiness and dysphasia. The initial CT of endocranium demonstrated haemorrhagic content. The patient received emergency treatment of 90 microg/kg/2h rFVIIa (NovoSeven) twice. The following day, after the first signs of ICH appeared, the patient developed tonic-clonic seizures characterized as epilepsy, which requiring antioedematous and anticonvulsive therapy. During the seizure, the patient bit his tongue, which resulted in massive bleeding from the wound and haematoma formation at the oral cavity floor. The first signs of resolution of ICH appeared three days after the first epileptic attack, and after following 11 days all signs completely withdrew. The patient received 29 doses of rFVIIa every 2 hours until the first signs of resolution of ICH and until bleeding from the injured tongue stopped. In the meantime, the patient had an episode of massive haematuria, which was successfully stopped with three doses of rFVIIa, 4.8 mg each. The titer of inhibitors was in the range of up to 26 units during hospitalization. The total dose of rFVIIa administered for treating ICH, tongue lesion, and haematuria was 153.6 mg. The patient also received 1 mg/kg of prednisone for three weeks for eradication of the inhibitors, and afterwards the dose was reduced to 0.5 mg/kg for the following month. CONCLUSION: If there is suspicion of ICH in haemophilic patients, it is necessary to administer replacement treatment, and if haemophilia with inhibitor is in question, as in our patient, immediate administration of rFVIIa is necessary followed by diagnostic procedures and the verification of ICH to determine further treatment and the dosage of rFVIIa, with clinical monitoring using radiological imaging. PMID- 19562873 TI - [Treatment of psoas haematoma in a patient with haemophilia and inhibitors]. AB - INTRODUCTION: Particular problem in treating haemophiliacs with serious muscle bleeding such as iliopsoas haematoma, are patients with inhibitor. CASE OUTLINE: Our study describes three episodes of psoas haematoma in a patient with haemophilia and inhibitor successfully treated with recombinant activated factor VII (rFVIIa). CONCLUSION: Based on our experience, initiating therapy within two hours in our patient, contributed to successful treatment, although small cumulative doses of rFVIIa were used. Therefore, introducing home treatment along with adequate patient and family education similar to developed countries becomes imperative in our environment, not only for providing better quality of life, but also from pharamaco-economic point of view. PMID- 19562874 TI - [Rehabilitation in haemophilic children with inhibitors using recombinant activated factor VII]. AB - INTRODUCTION: During invasive and orthopaedic procedure of the joints, physical therapy is a necessary modality for successful recovery, especially in patients with haemophilia. These patients require concurrent substitution therapy. Recombinant activated factor VII (rFVIIa) has been successfully used in treating haemophilia with inhibitor, especially during preoperative preparation. Nevertheless, optimal substitution therapy during perioperative physical therapy has been poorly defined. CASE OUTLINE: In our paper, we review the literature and the course of perioperative rehabilitation after synovectomy in a 16-year-old haemophilic patient with high-titre inhibitor, treated with rFVIIa. Rehabilitation was started on day 3 after surgery, with rFVIIa substitution lasting ten days and followed by intermittent dosages during episodes of pain. After concluding physical treatment, a significantly better function of the operated joint was recorded, which led to the recovery of walking without orthopaedic aid. CONCLUSION: Patients with inhibitors require individualized approach of a multidisciplinary team in order to achieve optimal results in elective orthopaedic procedures. PMID- 19562875 TI - [The use of recombinant activated factor VII to control haemorrhage following exenteration and massive transfusion in coagulopathic patients]. AB - INTRODUCTION: Exentheresis pelvis totalis due to locally advanced pelvic malignancies is an extensive surgical procedure. The operation is commonly associated with anticipated perioperative haemorrhage requiring a large volume of haemoproducts. Sometimes, the intervention can result in unexpected massive and uncontrolled haemorrhage that is frequently a combination of surgical and coagulopathic bleeding. Attempts to arrest massive bleeding by conventional means may fail. CASES OUTLINE: We describe our experience in the use of recombinant activated factor VII (rFVIIa) in three previously hemostatically competent patients who underwent exentheresis in order to control massive bleeding resulting from dilution coagulopathy in the operating theatre, as well as in the treatment of postoperative bleeding associated with consumptive coagulopathy. Of these, two massively transfused patients developed dilution coagulopathy in the operative theatre, which was poorly responsive to conventional management. In both cases, a single dose of rFVIIa (70 microg/kg and 60 microg/kg respectively) was given. Prompt clinical response was achieved and operations were successfully finished. In the third case, the patient developed consumptive coagulopathy on the first day after surgical procedure that was treated with conventional therapy. On the second postoperative day the patient became anuric and experienced severe intraabdominal bleeding. The bleeding was successfully controlled with rFVIIa in a single dose of 70 microg/kg. CONCLUSION: RFVIIa can be a treatment option in patients suffering from intractable coagulopathic bleeding when standard therapy has failed. PMID- 19562876 TI - [The use of recombinant activated factor VII in the treatment of gastrointestinal bleeding following acetylsalicylic acid therapy in a surgical patient]. AB - INTRODUCTION: Gastrointestinal bleeding is the most important complication associated with acetylsalicylic acid therapy. Patients with preexisting haemostatic disorders are at the higher risk and may experience life-threatening hemorrhagic syndrome. Platelet transfusions and desmopressin administration commonly successfully arrest bleeding. However, in clinical situations with profound bleeding and haemorrhagic shock, these therapeutic approaches may fail. CASE OUTLINE: We report a 24-year old female patient with previously undetected acquired platelet dysfunction, who underwent reconstructive surgical intervention. On the 20th postoperative day, acetylsalicylic acid was introduced due to reactive thrombocytosis (platelet count 1480x10(9)/L) with daily dose of 100 mg tablets. On the 12th day of the acetylsalicylic acid treatment, massive gastrointestinal bleeding with haemorrhagic shock suddenly occurred. Attempts to control massive haemorrhage by resuscitation, blood products and haemostatics (desmopressin, tranexamic acid) failed. Two bolus doses of recombinant activated factor VII (rFVIIa) (100 microg/kg and 60 microg/kg respectively) in 90 minutes interval were given. Bleeding was successfully controlled with no requirements for further haemoproducts and haemostatic remedies treatment. CONCLUSION: This case demonstrates that the use of rFVIIa may be a specific treatment option in patients suffering from severe gastrointestinal bleeding associated with acetylsalicylic acid treatment. PMID- 19562877 TI - [The use of recombinant activated factor VII in a patient with uncontrolled bleeding from the gastrointestinal tract]. AB - INTRODUCTION: Massive intestinal bleeding is a life threatening clinical condition that requires prompt reaction. Recombinant activated factor VII (rFVIIa) is a novel and promising therapeutic approach. The administration of rFVIIa usually represents the last therapy resort, when all other alternatives are exhausted. CASE OUTLINE: We present a 55-year old female patient admitted to our hospital due to intestinal bleeding of unknown origin, with anamnestic and heteroanamnestic data, clinical condition that was the consequence of massive bleeding, diagnostic and therapeutic procedures. After diagnostic procedures, the conventional medicamentous therapy was commenced. On the 6th day of therapy, surgery was performed with the aim to identify the source of bleeding. After the confirmation that the bleeding was not due to surgical reasons, we administrated rFVIIa in a bolus dose of 90 mcg/kg, which resulted in the prompt arrest of bleeding. Side effects of the drug were not registered. CONCLUSION: In our patient, the administration of rFVIIa resulted in a prompt and efficient haemostasis, which was not followed by the recurrence of bleeding. Preconditions necessary for rFVIIa administration are the adequate substitution of blood products, exclusion of surgical cause of bleeding, as well as the levels of fibrinogen, platelets and pH equal or above recommended. PMID- 19562878 TI - [Intraoperative use of recombinant activated factor VII in traumatic unpenetrating liver injury--case report]. AB - INTRODUCTION: Uncontrolled massive bleeding is often the cause of death of polytraumatized patients. Massive haemorrhage in polytrauma is the consequence of severe tissue and blood vessel damage or the development of posttraumatic coagulopathy. Most often, it is the combination of the two causes. Coagulopathy arises in early stages of trauma and it is an independent predictor of mortality of polytraumatized patients, however, its timely correction can significantly result in the reduction of mortality rate in trauma. CASE OUTLINE: We present a 15-year old male patient who fell from 11 m height and sustained injuries of the right lobe of liver, with partial avulsion and lacerations on the diaphragmal and visceral side of the right lobe. He also sustained brain injuries with presence of blood content in the right lateral chamber, numerous haemorrhagic spots and contusions in frontoparietal region and diffuse brain oedema of the right side of the hemisphere. A partial right sided hepatectomy was performed using the surgical techniques of haemostasis (ligation, tamponade, compression, embolisation), circulatory volume resuscitation (crystalloids and colloids), resuscitation of blood and blood components and the application of antifibrynolitics, as well as oxigenotherapy (endotracheal intubation and mechanical ventilation, with high inspiratory fractions of oxygen). Considering massive transfusions, the type and character of injury and visible oozing, as well as the fact that conventional methods did not stop the bleeding, recombinant activated factor VII (rFVIIa) was given intraoperatively, which resulted in successful reduction and finally the cecession of bleeding. CONCLUSION: Until today, there are numerous references about successful application of rFVIIa in uncontrolled bleedings in trauma when previously applied conventional methods of haemostasis were not sufficient. We are presenting a case of successful use of rFVIIa in our hospital accompanied by usual surgical measures and reanimation of a severely injured patient with massive bleeding. Based on our experience and available references, in case of timely diagnosis and adequate therapy, such as conventional treatment methods and the use of rFVIIa, uncontrolled bleedings in polytrauma have better prognosis. PMID- 19562879 TI - [Successful application of recombinant activated factor VII in a patient with HELLP syndrome and disseminated intravascular coagulation]. AB - INTRODUCTION: HELLP syndrome represents the form of preeclampsia characterized by moderate hypertension, often with absence of proteinuria and oedema. The frequency of HELLP syndrome in pregnant women with preeclampsia is 10-20%. The clinical course of the disease is characterized by the progressive worsening of mother and fetus condition, which can be stopped only by delivery. Disseminated intravascular coagulation is present in 8% of patients with HELLP syndrome and causes significant morbidity and mortality. CASE OUTLINE: We present a case of HELLP syndrome complicated by intrauterine fetal demise and disseminated intravascular coagulation in trigemelar pregnancy. After all surgical and medicamentous methods to establish haemostasis were exhausted, the patient was treated by recombinant activated factor VII (rFVIIa) in intravenous bolus dose of 90 microg/kg twice, which resulted in satisfactory haemostasis. Side effects of the drug were not registered. CONCLUSION: The application of rFVIIa reduced haemorrhage in our patient, both after the Caesarean section and after hysterectomy, contributing to the patient's full recovery, without neurological sequelae and with preserved renal function. RFVIIa is not an alternative to surgical haemostasis, but its administration should surely be considered before deciding to perform hysterectomy, especially in patients who want to preserve fertility. In cases of postpartum haemorrhage, when bleeding persists even after adequate surgical haemostasis, the administration of rFVIIa is to be considered not only as an alternative to hysterectomy, but also an effort to prevent significant maternal morbidity and mortality. PMID- 19562880 TI - [Recombinant activated factor VII in haemostasis after radical nephrectomy]. AB - INTRODUCTION: The paper presents important facts in the application of recombinant human factor VIIa (rFVIIa) and in the treatment of renal cell carcer. CASE REPORT: A 69-year old male with infiltrative renal cell carcer underwent radical nephrectomy using Hasagawa's approach. The extirpated tumor was 35 cm in diameter. During surgery, the patient suffered cardiac arrest with 3500 ml blood loss. Twenty-four hours after operation, he lost additional 2100 ml of blood. Despite adequate blood substitution with intensive surgical attempts to treat haemostasis, bleeding did not stop until the application of rFVIIa (300 IU). The patient survived and fully recovered. CONCLUSION: To control massive non-surgical bleeding additionally complicated by paraneoplastic syndrome, extensive surgical intervention, haemodilution and massive blood substitution, the application of rFVIIa has proved to be most successful. PMID- 19562881 TI - Role of physiologically balanced growth factors in skin rejuvenation. PMID- 19562882 TI - Topically applied physiologically balanced growth factors: a new paradigm of skin rejuvenation. AB - Synergistic interaction of multiple growth factors (GF) in skin controls the processes that promote skin repair. GFs have been shown to affect different pathways of skin repair and rejuvenation with many GFs working in close cooperation with one another and with other endogenous agents. Intrinsic and extrinsic aging of skin reduces both the levels of natural GFs and the number and activity of fibroblasts. Supplementing skin's endogenous GFs may enhance natural repair processes and accelerate the reversal of damage caused by intrinsic and extrinsic skin aging. In spite of their large molecular weight, evidence suggests that a small fraction of topically applied GFs penetrating into superficial epidermis can elicit a fibroblast-mediated response in the dermis. GF mixture secreted by human fibroblasts grown in conditions resembling the physiological condition of dermis, and present at high concentrations in a stable formula is most likely to provide an ideal cosmeceutical product. This naturally balanced mixture is also likely to contain other important, but as yet unidentified, substances that affect skin healing. Such a complex mixture cannot be reproduced using synthetic substances. Clinical studies have shown that topical application of products containing high concentrations of a physiologically balanced mixture of GF appears to reverse the signs of skin aging. A synergistic combination of antioxidants, matrix building agents and skin conditioners with physiologically balanced GF provides a novel and comprehensive paradigm of skin rejuvenation. PMID- 19562883 TI - "Natural" ingredients in cosmetic dermatology. AB - Recently, both clinical and bench research has begun to provide scientific validation for the use of certain botanical ingredients. Related findings regarding proposed biological mechanisms of action have translated into clinical practice. Botanical compounds for which dermatologic and cosmetic applications have emerged include: olive oil, chamomile, colloidal oatmeal, oat kernal extract, feverfew, acai berry, coffee berry, curcumin, green tea, pomegranate, licorice, paper mulberry, arbutin, and soy. Many of these botanical sources offer biologically active components that require further in vitro and in vivo investigation in order for us to properly educate ourselves, and our patients, regarding over-the-counter products based on these ingredients. PMID- 19562884 TI - Optimizing patient care with "natural" products: treatment of hyperpigmentation. AB - Patients with skin of color suffer from different cutaneous issues when compared with skin of light complexion, and therefore management of the former must be representative of these variations. The most common pigmentary complaints in patients with skin of color are post-inflammatory hyperpigmentation, melasma and sun-induced hyperpigmentation. Often, patients with darker skin will turn to naturally occurring ingredients over synthetic analogues both to satisfy cultural preferences and to limit potential adverse effects that have been tied to synthetics. Science-based natural products can offer an attractive adjunct to conventional therapies that treat melasma, post-inflammatory hyperpigmentaion, and other dyschromias. Increasing data on the biological effects and the efficacy of natural therapies support the use of these complementary therapies in treating hyperpigmentation. PMID- 19562885 TI - What is the standard of care for anesthesia? Who determines it? PMID- 19562887 TI - Use of masseteric and deep temporal nerve blocks for reduction of mandibular dislocation. AB - A patient presented with a unilateral dislocated condyle that was resistant to reduction by simple manual manipulation because of elevator muscle spasm and severe muscle and temporomandibular joint pain. A technique involving a masseteric nerve block and a temporal nerve block was used, allowing a quick, safe, and minimally painful reduction. The method used for delivering these nerve blocks is described here. PMID- 19562886 TI - Does granisetron eliminate the gag reflex? A crossover, double-blind, placebo controlled pilot study. AB - Although gagging is a frequent problem that, when severe, can jeopardize the dental procedure, no single protocol is used to alleviate this phenomenon. Selective 5-HT3 antagonists, such as granisetron, may attenuate gagging. In this study, granisetron and placebo were administered intravenously, in a crossover, double-blind manner, to 25 healthy volunteers in 2 different sessions. Gagging levels were recorded before and after administration, as were BP, pulse, and O2 saturation. Recorded results were analyzed with the use of tests for nonparametric values (P = .05). A significant increase in the depth of swab insertion was noted after administration of both placebo and drug. The increase in drug effectiveness correlated with decreased body weight. The true efficacy of granisetron in gagger patients with this treatment protocol has yet to be fully established, although it has been theorized that an increased dosage of granisetron may have a better effect. PMID- 19562888 TI - Respiratory monitoring: physiological and technical considerations. AB - The American Dental Association and several dental specialty organizations have published guidelines that detail requirements for monitoring patients during various levels of sedation and, in some cases, general anesthesia. In general, all these are consistent with those guidelines suggested by the American Society of Anesthesiologists for sedation and analgesia by nonanesthesiologists. It is well accepted that the principal negative impact of sedation and anesthesia is the compromise of respiratory function. While monitoring per se is a technical issue, an appreciation of its purpose and the interpretation of the information provided require an understanding of respiratory anatomy and physiology. The focus of this continuing education article is to address the physiological aspects of respiration and to understand the appropriate use of monitors, including the interpretation of the information they provide. PMID- 19562890 TI - Case reports versus evidence-based medicine (EBM). PMID- 19562889 TI - Guideline for the diagnosis, prevention and treatment of paediatric ventilator associated pneumonia. AB - OBJECTIVE: Ventilator-associated pneumonia (VAP) has been poorly studied in South Africa, but is likely to be a significant problem, with resulting increased morbidity and mortality in the paediatric intensive care unit population. This guideline is intended to review the evidence and recommendations for prevention and management of VAP in children and to provide, where possible, clear advice to aid the care of these children, to limit costly and unnecessary therapies and- importantly--limit inappropriate use of antimicrobial agents, EVIDENCE: The Working Group was constituted. Literature on the aetiology, prevention and management of paediatric VAP is reviewed. RECOMMENDATIONS: Evidence-based clinical practice guidelines are provided for VAP diagnosis and prevention in South Africa. In addition, the current status of antimicrobial use has been reviewed and clear recommendations are set out. PMID- 19562891 TI - Recurrent laryngeal neuropathy (RLN) research: where are we and to where are we heading? PMID- 19562892 TI - Effect of magnitude and direction of force on laryngeal abduction: implications for the nerve-muscle pedicle graft technique. AB - REASONS FOR PERFORMING STUDY: The nerve-muscle pedicle graft technique is a treatment for recurrent laryngeal neuropathy (RLN), but the optimal placement of the pedicles within the cricoarytenoideus dorsalis (CAD) muscle is unknown. HYPOTHESIS: The magnitude and direction of force placed on the muscular process of the left arytenoid cartilage affects the magnitude of laryngeal abduction. METHODS: Five larynges were harvested from cadavers. Using increments of 0.98 N, a dead-weight force generator applied a force of 0-14.7 N for 1 min each to the left muscular process at 0, 10, 20, 30, 40, 50, 60 and 70 degrees angles. The rima glottis was photographed digitally 1 min after each force had been applied. Distances between biomarkers (Lines 1-4) and right to left angle quotient (RLQ) were used to assess the degree of left arytenoid abduction. RESULTS: Increasing force from 0-14.7 N progressively and significantly increased the length of all lines and RLQ, indicating abduction. Furthermore, there was a significant interaction between force and angles. Applying forces of 7.84 N or greater (Lines 2-4 and RLQ) or 11.76 N or greater (Line 1) at angles 0, 10, 20 and 30 degrees resulted in significantly greater abduction than applying the same forces at 40, 50, 60 and 70 degrees. Angles of 0-30 degrees correspond with the direction of pull exerted by the lateral compartment of the CAD muscle. CONCLUSION: In RLN, nerve-muscle pedicle grafts should be placed preferentially in the lateral rather than in the medial compartment of the CAD muscle. POTENTIAL RELEVANCE: The information presented can be used to assist surgeons in the planning and application of the nerve-muscle pedicle graft procedure. PMID- 19562893 TI - The effect of focused extracorporeal shock wave therapy on collagen matrix and gene expression in normal tendons and ligaments. AB - REASONS FOR PERFORMING STUDY: Extracorporeal shock wave therapy (ESWT) is frequently used in equine practice, but little is known about its biological action. OBJECTIVES: To study the effects of ESWT on matrix structure and gene expression levels in normal, physiologically loaded tendinous structures in ponies. METHODS: Six Shetland ponies, free of lameness and with ultrasonographically normal flexor and extensor tendons and suspensory ligaments (SL), were used. ESWT was applied at the origin of the suspensory ligament and the mid-metacarpal region of the superficial digital flexor tendon (SDFT) 6 weeks prior to sample taking, and at the mid-metacarpal region (ET) and the insertion on the extensor process of the distal phalanx (EP) of the common digital extensor tendon 3 h prior to tendon sampling. In all animals one forelimb was treated and the other limb was used as control. After euthanasia, tendon tissue was harvested for real-time PCR to determine gene expression levels and additional samples were taken for histological evaluation and biochemical analyses RESULTS: Histologically a disorganisation of the normal collagen structure was observed 3 h after ESWT, remnants of which were still visible after 6 weeks. While degraded collagen levels showed an increase at 3 h post treatment (P= 0.012) they were reduced at 6 weeks post ESWT (P = 0.039). Gene expression for both COL1 (P = 0.004) and MMP14 (P = 0.020) was upregulated at 6 weeks after treatment. CONCLUSIONS: Exposure of normal tendinous tissue to ESWT is not uneventful; it leads to a disorganisation of matrix structure and changes in degraded collagen levels. The upregulation of COL1 expression 6 weeks after ESWT may be indicative for repair. POTENTIAL RELEVANCE: The observed disorganisation of the collagen network warrants caution when using ESWT. Exposing noninjured tissue to ESWT should be avoided and it may be advisable to restrict exercise in recently treated patients. However, the induced tissue disorganisation might also be a trigger for repair in chronic tendinopathies. PMID- 19562894 TI - Variability of resting endoscopic grading for assessment of recurrent laryngeal neuropathy in horses. AB - REASONS FOR PERFORMING STUDY: The extent to which variability affects endoscopic grading of arytenoid cartilage movement is uncertain. OBJECTIVE: To determine the observer and within horse variability of grading arytenoid cartilage movement in horses during resting endoscopic examination, using a 7-grade system. METHODS: Endoscopic recordings of the upper respiratory tract made at rest in 270 draught horses were reviewed independently by 2 veterinarians to assess interobserver variability when scoring horses' laryngeal function with a 7-grade system. Grading was repeated by both examiners in 80 randomly selected recordings in order to assess intraobserver variability. In 120 horses, endoscopy was repeated after 24-48 h, with videos graded by both veterinarians to assess intrahorse variability. RESULTS: The mean weighted kappa statistic for concordance within examiners was 0.867, with a mean intraobserver agreement of 763%. The weighted kappa statistic for concordance between the 2 examiners was 0.765, with an interobserver agreement of 63.1%. Of the horses receiving 2 endoscopic examinations, the same grade was assigned to 57.1% of horses at the second examination, when effects resulting from interobserver variability were removed. The mean weighted kappa statistic for concordance between the grade assigned at first vs. second examinations was 0.588, indicating only moderate agreement. CONCLUSIONS AND POTENTIAL RELEVANCE: Intra- and interobserver reliability of resting endoscopic grading of arytenoid cartilage movement using a 7-grade system was high when examinations were conducted by experienced veterinarians. However, there was moderate daily intrahorse variability, suggesting that results of resting endoscopic examinations performed on a single day should be interpreted with caution. PMID- 19562895 TI - Dynamic respiratory endoscopy without treadmill in 68 performance Standardbred, Thoroughbred and saddle horses under natural training conditions. AB - REASONS FOR PERFORMING STUDY: High speed treadmill endoscopy provides a true assessment and diagnosis of the dynamic obstructions of the upper equine respiratory tract (DO-URT). However, treadmills do not always allow reproduction of the exact conditions of dynamic collapse of the URT. The availability of on board endoscopes, which could be used without a treadmill, would make exercising endoscopy readily available to more equine practices. OBJECTIVES: To develop and validate an innovative endoscope which could be used for the examination of the URT at exercise in ridden and harnessed sport horses performing in natural conditions. METHODS: Authors worked closely with engineers of a company manufacturing veterinary endoscopes. Over a 2-year period several prototypes were tested on more than 20 cooperative horses performing either in trot, gallop, jumping or endurance. The final Dynamic Respiratory Scope (DRS) allows real-time visualisation of the URT and video recordings for post test reviewing and archiving. The DRS was then tested in 2 equine livery yards. Performance horses presented for investigation of abnormal respiratory noises at exercise and/or poor performance were subjected to exercising endoscopy. Endoscopic images of the URT were recorded and video recordings of the URT were reviewed post test. RESULTS: A total of 68 horses were examined: 39 harnessed Standardbred and 29 mounted horses (16 Thoroughbred and 13 saddle horses). Of these, 44 were diagnosed with a URT abnormality. Good quality videos were obtained even at maximum speed in all cases. The innovative insertion tube provided very stable images compared to those obtained with flexible video endoscopes on treadmills. CONCLUSIONS: This study validates the safety and the reliability of the DRS for imaging the equine URT during natural exercising conditions. POTENTIAL RELEVANCE: Most common causes of DO-URT can be diagnosed easily during any type of performance without a treadmill and the DRS offers a great potential for further URT clinical research. PMID- 19562896 TI - Dynamic respiratory endoscopy in 67 Thoroughbred racehorses training under normal ridden exercise conditions. AB - REASONS FOR PERFORMING STUDY: There are potential advantages to imaging the upper portion of the respiratory tract (URT) of horses during ridden exercise. With the advent of a wireless endoscope, this is now possible. However, there has been no detailed validation of the technique and findings have not been compared to ridden speed. OBJECTIVES: To assess the combined use of a Dynamic Respiratory Scope (DRS) and global positioning system (GPS) receiver for examining the URT of a group of Thoroughbred racehorses randomly selected from a single flat racehorse training establishment. METHODS: Horses were selected randomly from a population of Thoroughbred horses in training at the same training yard. Endoscopic images of the URT were recorded during a ridden exercise test on an 'all-weather-gallop' and were reviewed post testing. Speed was measured using a wrist mounted GPS receiver. RESULTS: A total of 67 (34%) of the 195 horses in training were examined. Endoscopic findings included: normal URT function (44 cases); dorsal displacement of the soft palate (DDSP) (13 cases); laryngeal asymmetry (4 cases); and axial deviation of the aryepiglottic folds (3 cases). Maximum speed obtained by individual horses ranged from 41.8-56.3 km/h. Ridden speed was variably affected by DDSP. CONCLUSIONS: The DRS provides a safe effective system for imaging the equine URT during ridden exercise at speed. The abnormalities of the URT identified were similar to those observed during treadmill endoscopy studies reported in the literature. The effect of URT abnormalities on ridden speed requires further investigation. POTENTIAL RELEVANCE: This technique can be used to diagnose common causes of URT associated with poor performance in horses during normal training. This has substantial implications for future clinical diagnosis and treatment of URT pathology. PMID- 19562897 TI - Pharmacokinetics of detomidine and its metabolites following intravenous and intramuscular administration in horses. AB - REASONS FOR PERFORMING STUDY: Detomidine is commonly used i.v. for sedation and analgesia in horses, but the pharmacokinetics and metabolism of this drug have not been well described. OBJECTIVES: To describe the pharmacokinetics of detomidine and its metabolites, 3-hydroxy-detomidine (OH-detomidine) and detomidine 3-carboxylic acid (COOH-detomidine), after i.v. and i.m. administration of a single dose to horses. METHODS: Eight horses were used in a balanced crossover design study. In Phase 1, 4 horses received a single dose of i.v. detomidine, administered 30 microg/kg bwt and 4 a single dose i.m. 30 microg/kg bwt. In Phase 2, treatments were reversed. Plasma detomidine, OH detomidine and COOH-detomidine were measured at predetermined time points using liquid chromatography-mass spectrometry. RESULTS: Following i.v. administration, detomidine was distributed rapidly and eliminated with a half-life (t1/2(el)) of approximately 30 min. Following i.m. administration, detomidine was distributed and eliminated with t1/2(el) of approximately one hour. Following, i.v. administration, detomidine clearance had a mean, median and range of 12.41, 11.66 and 10.10-18.37 ml/min/kg bwt, respectively. Detomidine had a volume of distribution with the mean, median and range for i.v. administration of 470, 478 and 215-687 ml/kg bwt, respectively. OH-detomidine was detected sooner than COOH detomidine; however, COOH-detomidine had a much greater area under the curve. CONCLUSIONS AND POTENTIAL RELEVANCE: These pharmacokinetic parameters provide information necessary for determination of peak plasma concentrations and clearance of detomidine in mature horses. The results suggest that, when a longer duration of plasma concentration is warranted, the i.m. route should be considered. PMID- 19562898 TI - Post mortem evaluation of palmar osteochondral disease (traumatic osteochondrosis) of the metacarpo/metatarsophalangeal joint in Thoroughbred racehorses. AB - REASONS FOR PERFORMING STUDY: Thoroughbred racehorses are commonly affected by subchondral bone injury, but the exact prevalence and the distribution of palmar/plantar osteochondral disease (POD) lesions are unknown. The relationship between pathologies has not been elucidated, although it is widely accepted that POD is a manifestation of traumatic overload arthrosis. HYPOTHESIS: There is an association between grade of POD and other pathologies affecting the third metacarpal and metatarsal (MC/MTIII) condyles (wear lines, cartilage loss, marginal remodelling, dorsal impact injuries and linear fissures). OBJECTIVES: To evaluate the pathology found affecting the distal MC/MTIII condyles of Thoroughbred racehorses at post mortem examination, to describe the prevalence and distribution of POD lesions within a population of racing Thoroughbreds and to determine relationships between pathologies of the distal condyles of the third metacarpal and metatarsal bones. METHODS: The metacarpo/metatarsophalangeal joints of 64 Thoroughbred racehorses were examined at routine post mortem examination and graded for third metacarpal and metatarsal condylar pathology. Associations between pathologies were determined. RESULTS: POD had a within horse prevalence of 67%. There was a significant linear relationship between grade of POD and grades of wear lines, cartilage ulceration and dorsal impact injuries. There was a significant relationship, but this was not linear, between grade of POD and grade of linear fissures. Using ordinal logistic regression, compared to condyles with grade 0 or grade 2 linear fissures, condyles with grade 1 linear fissures were found to be more likely to have a lower POD grade. POTENTIAL RELEVANCE: POD can be considered to be a manifestation of traumatic overload arthrosis, but the role of subchondral bone adaptation is complex and warrants further investigation. PMID- 19562899 TI - Descriptive epidemiology of fracture, tendon and suspensory ligament injuries in National Hunt racehorses in training. AB - REASONS FOR PERFORMING STUDY: While fractures and tendon injuries are known to be important diseases in National Hunt (NH) racehorses during racing, there are no accurate estimates of their incidence in NH training yards. OBJECTIVES: To estimate the incidence of fractures and tendon and suspensory ligament injuries (TLIs) in NH racehorses in training; to describe the injuries incurred and to compare injury incidence rates by horse age, trainer, gender and background (ex flat vs. ex-store horses). METHODS: Cohort data were collected from 14 UK NH training yards for 2 racing seasons. Daily exercise regimens and details of fractures and TLIs occurring in study horses were recorded. RESULTS: Data were gathered from 1223 horses that spent 9466 months at risk of injury. The fracture incidence rate was 1.1/100 horse months and varied significantly by trainer (P<0.001) but not by gender, age or background. The pelvis and third metacarpal bone (MCIII) were the most common fracture sites, although this varied between racing and training. The TLI incidence rate was 1.9/100 horse months and varied significantly by trainer (P = 0.05) and age (P<0.001) but not by gender or background. However, ex-store horses were significantly more likely to have a TLI on the racecourse than ex-flat horses (P = 0.01). Superficial digital flexor injuries accounted for 89% of all TLIs, the remainder being suspensory ligament injuries. CONCLUSIONS AND POTENTIAL RELEVANCE: Fractures and TLIs are important causes of morbidity and mortality in NH racehorses in training in England. This study provides accurate estimates of their incidence in this population and provides a baseline against which to monitor the effect of future interventions. PMID- 19562900 TI - Diffusion of contrast medium after perineural injection of the palmar nerves: an in vivo and in vitro study. AB - REASONS FOR PERFORMING STUDY: Proximal diffusion of local anaesthetic solution after perineural anaesthesia may lead to the desensitisation of structures other than those intended. However, there is no evidence-based study demonstrating the potential distribution and diffusion of local anaesthetic solution after perineural analgesia in the distal limb. OBJECTIVE: To document the potential diffusion of local anaesthetic solution using a radiopaque contrast model and to evaluate the influence of walking compared with confinement in a stable after injection. METHODS: Radiopaque contrast medium was injected subcutaneously over one palmar nerve at the base of the proximal sesamoid bones in 6 nonlame mature horses. Horses were assigned randomly to stand still or walk after injection. Radiographs were obtained 0, 5, 10, 15, 20 and 30 min after injection and were analysed to determine the distribution and diffusion of the contrast medium. RESULTS: In 89% of injections an elongated pattern of the contrast medium was observed suggesting distribution along the neurovascular bundle. After 49% of injections a fine radiopaque line extended proximally from the contrast 'patch', and in 25% of injections a line extended distally. There was significant proximal and distal diffusion with time when sequential radiographs of each limb were compared. The greatest diffusion occurred in the first 10 min. Walking did not significantly influence the extent of either proximal or distal diffusion. CONCLUSIONS AND POTENTIAL RELEVANCE: Significant proximal diffusion occurs in the first 10 min after perineural injection in the distal aspect of the limb and should be considered when interpreting nerve blocks. Distribution of local anaesthetic solution outside the fascia surrounding the neurovascular bundle or in lymphatic vessels may explain delayed or decreased effects. PMID- 19562901 TI - Influence of subclinical inflammatory airway disease on equine respiratory function evaluated by impulse oscillometry. AB - REASONS FOR PERFORMING STUDY: Inflammatory airway disease (IAD) is a nonseptic condition of the lower respiratory tract. Its negative impact on respiratory function has previously been described using either forced expiration or forced oscillations techniques. However, sedation or drug-induced bronchoconstriction were usually required. The impulse oscillometry system (IOS) is a noninvasive and sensitive respiratory function test validated in horses, which could be useful to evaluate IAD-affected horses without further procedures. OBJECTIVES: To determine the sensitivity of IOS in detecting alterations of the respiratory function in subclinically IAD-affected horses without inducing bronchoprovocation and to characterise their respiratory impedance according to frequency for each respiratory phase. METHODS: Pulmonary function was evaluated at rest by IOS in 34 Standardbred trotters. According to the cytology of bronchoalveolar lavage fluid (BALF), 19 horses were defined as IAD-affected and 15 horses were used as control (CTL). Total respiratory resistance (Rrs) and reactance (Xrs) from 1-20 Hz as well as their inspiratory and expiratory components were compared between groups. RESULTS: A significant increase of Rrs at the lower frequencies (R1-10 Hz) as well as a significant decrease of Xrs beyond 5 Hz (X5-20 Hz) was observed in IAD compared to CTL horses. IOS-data was also significantly different between inspiration and expiration in IAD-affected horses. In the whole population, both BALF eosinophil and mast cell counts were significantly correlated with IOS measurements. CONCLUSIONS: Functional respiratory impairment may be measured, even in the absence of clinical signs of disease. In IAD-affected horses, the different parameters of respiratory function (Rrs or Xrs) may vary depending on the inflammatory cell profiles represented in BALF. POTENTIAL RELEVANCE: Impulse oscillometry could be used in a routine clinical setting as a noninvasive method for early detection of subclinical respiratory disease and of the results of treatment in horses. PMID- 19562902 TI - Clinical dental examinations of 357 donkeys in the UK. Part 1: prevalence of dental disorders. AB - REASONS FOR PERFORMING STUDY: Dental disorders have a high prevalence in older donkeys and horses, but the nature and pathogenesis of many of these disorders have yet to be established. OBJECTIVES: The identification and determination of the prevalence of important dental disorders in different age groups in a large single population of donkeys, to establish a better understanding of the nature and pathogenesis of these disorders. METHODS: A prospective cross-sectional study was performed on the donkey population at The Donkey Sanctuary with detailed oral examinations of 357 donkeys within 7 different age groups (age range 2-53 years) recorded. RESULTS: The prevalence of dental disease in all donkeys was 73%, increasing in prevalence from 28% in the youngest to 98% in the oldest age group. There was an increase in prevalence of commonly recognised dental disorders with increasing age, such as: diastemata (3.8% in youngest to 86% in oldest group); missing teeth (0-56%); overgrown teeth (15-86%); worn teeth (8-84%); displaced teeth (0-38%); and periodontal disease (0-28%). CONCLUSIONS: There was a significant increase in the prevalence of dental disorders with increasing age with the largest significant increase for most dental disorders occurring in the 15-20 year age group. POTENTIAL SIGNIFICANCE: Most dental disorders significantly increase in prevalence in the 15-20 year age group and, therefore, prophylactic geriatric dental treatment in donkeys should be commenced from age 15 years. PMID- 19562903 TI - Clinical dental examinations of 357 donkeys in the UK. Part 2: epidemiological studies on the potential relationships between different dental disorders, and between dental disease and systemic disorders. AB - REASONS FOR PERFORMING STUDY: Dental disease has been shown to be a risk factor for weight loss and colic in horses. No extensive clinical studies in donkeys have investigated the potential relationship between different dental disorders, or between dental disease and systemic disorders. OBJECTIVES: To determine possible associations between dental disease and body condition score, weight loss, the need for supplemental feeding and prevalence of colic in donkeys of all ages, and to gain a better understanding of the pathogenesis of dental disease by the determination of associations between different dental disorders. METHODS: A prospective cross-sectional analysis of clinical dental examinations of 357 donkeys in The Donkey Sanctuary, Sidmouth was performed. Other epidemiological factors such as estimated age group, body condition score, weight loss, medical history and supplemental feeding were also recorded, and multiple regression analyses were performed to determine possible associations. RESULTS: Donkeys from older age groups were more likely to have dental disease, poor body condition score and suffered previous colic episodes. The presence of dental disease was also significantly associated with weight loss, colic, low body condition score and the need for supplemental feeding. The presence of diastemata, periodontal disease, wave mouth, smooth mouth and step mouth are frequently associated with the presence of other dental disorders. CONCLUSIONS: In addition to oral-related pain, dental disease can cause significant systemic disorders and so has increased welfare implications in donkeys. Some dental disorders promote the development of other types of dental abnormalities and thus increase the severity of dental disease in individual animals. POTENTIAL SIGNIFICANCE: Effective treatment of dental disorders slows down the progression of dental disease and decreases the risk of developing some medical disorders such as colic and weight loss that are associated with dental disease. PMID- 19562904 TI - Retrospective multicentre study of methicillin-resistant Staphylococcus aureus infections in 115 horses. AB - REASONS FOR PERFORMING STUDY: Methicillin-resistant Staphylococcus aureus (MRSA) is an emerging veterinary and zoonotic pathogen, associated with increasing reports of disease in horses. OBJECTIVES: To provide an overview of the characteristics of clinical MRSA infections in horses. METHODS: A retrospective case study was performed on 115 horses admitted to 6 participating veterinary teaching hospitals in Canada and the United States between 2000 and 2006, and diagnosed with clinical MRSA infection. Descriptive statistics, univariate and multivariable analyses for community- (CA) vs. hospital-associated (HA) MRSA infections, and survival vs. nonsurvival at discharge were performed. RESULTS: The age range of MRSA-infected horses was zero (born in hospital) to 31 years. HA (58/114, 50.9%) and CA infections (56/114, 49.1%) were equally common. Infection of surgical incisions was most frequently reported (44/115, 38.0%). Overall 93/111 (83.8%) cases survived to discharge. Previous hospitalisation and treatment with gentamicin were associated significantly with CA-MRSA, whereas infected incision sites were associated significantly with HA-MRSA. Factors significantly associated with nonsurvival included i.v. catheterisation, CA-MRSA infection and dissemination of infection to other body sites. CONCLUSIONS: Equine MRSA infections have a broad range of clinical presentations, appear to be primarily opportunistic and the overall prognosis for survival to discharge is good. POTENTIAL RELEVANCE: These results should help direct future research with regard to investigation of risk factors for equine MRSA infection in community and hospital populations. PMID- 19562905 TI - Prevalence, number and morphological types of multinucleated histiocytic giant cells in equine inflammatory dermatoses: a retrospective light microscopic study of skin-biopsy specimens from 362 horses. AB - REASONS FOR PERFORMING STUDY: Multinucleated histiocytic giant cells (MHGC) are seen frequently in skin-biopsy specimens from horses with inflammatory dermatoses. However, the prevalence, number and morphological types of these cells have not been reported. OBJECTIVE: To determine the prevalence, number and morphological types of MHGC in equine inflammatory dermatoses, and the association of these cells with specific conditions. METHODS: Skin-biopsy specimens from 335 horses with inflammatory dermatoses and from 27 horses with normal skin were evaluated for the prevalence, number and morphological types of MHGC. RESULTS: The prevalence and number of MHGC were greater in granulomatous dermatoses than in nongranulomatous dermatoses. Infectious and noninfectious dermatoses were not different in terms of prevalence or morphological types of MHGC. Foreign-body MHGC were the predominant type in almost all cases. MHGC were not seen in normal skin. CONCLUSIONS: MHGC are seen in a wide variety of equine inflammatory dermatoses, especially those that are granulomatous. Number and morphological types of MHGC are of no apparent diagnostic significance. POTENTIAL RELEVANCE: MHGC are frequently present in a wide variety of inflammatory dermatoses in the horse. Because the prevalence, number and morphological types of MHGC are of minimal diagnostic significance, special stains and tissue cultures are necessary to confirm specific diagnoses. PMID- 19562906 TI - Bone biomarkers and risk of fracture in two- and three-year-old Thoroughbreds. AB - The aim of this study was to determine whether bone biomarkers (osteocalcin, PICP, ICTP and CTX-I) could be used to identify 2- and 3-year-olds at increased risk of fracture in the subsequent flat racing season. It was concluded that these bone biomarkers cannot be used to identify 2- and 3-year-olds that sustain a fracture. Whether bone biomarkers have better predictive value in older horses or when measured serially in the same animal remains to be determined. PMID- 19562907 TI - Social inequalities in adult female mortality by the National Statistics Socio economic Classification, England and Wales, 2001-03. AB - This analysis of mortality in women aged 25-59 in 2001-03 found that those in the least advantaged social economic class had a mortality rate around twice that of women in the most advantaged class. This article uses the National Statistics Socio-economic Classification (NS-SEC) and examines the relative merits of classification based on a woman's 'own' occupation as opposed to a 'combined' classification which also takes into account the husband's NS-SEC class, where available. The results demonstrate a strong socio-economic gradient in mortality for adult women under both classification methods. Under the 'combined' classification, women in the least advantaged NS-SEC class had a mortality rate 2.6 times that of those in the most advantaged class. Based on the women's'own' occupation, the comparable ratio was 1.9. These results set a benchmark for the future monitoring of socio-economic mortality inequalities in women, and also provide a comparison between inequalities affecting women and men. PMID- 19562908 TI - Multivariate analysis of infant death in England and Wales in 2005-06, with focus on socio-economic status and deprivation. AB - Current health inequality targets include the goal of reducing the differential in infant mortality between social groups. This article reports on a multivariate analysis of risk factors for infant mortality, with specific focus on deprivation and socio-economic status. Data on all singleton live births in England and Wales in 2005-06 were used, and deprivation quintile (Carstairs index) was assigned to each birth using postcode at birth registration. Deprivation had a strong independent effect on infant mortality, risk of death tending to increase with increasing levels of deprivation. The strength of this relationship depended, however, on whether the babies were low birthweight, preterm or small-for gestational-age. Trends of increasing mortality risk with increasing deprivation were strongest in the postneonatal period. Uniquely, this article reports the number and proportion of all infant deaths which would potentially be avoided if all levels of deprivation were reduced to that of the least deprived group. It estimates that one quarter of all infant deaths would potentially be avoided if deprivation levels were reduced in this way. PMID- 19562909 TI - An update to measuring chronic illness, impairment and disability in national data sources. AB - This article reports progress in delivering a revision to survey questions on disability for implementation carrying out in 2010. The Office for National Statistics has relied on survey data to report and update annual estimates of disability-free life expectancy at national level.The survey question used in this calculation has remained consistent since 1972. However, changes in national legislation, a forthcoming European regulation and a number of inadequacies in the level of detail and consistency of disability data collected in household surveys (raised as part of the Review of Equality Data), will require its modification and extension to ensure survey data better reflect the growing data needs in the subject area of disability in the 21st Century. The accurate and reliable measurement of disability is increasing in importance following the revision to the Disability DiscriminationActin 2005 and the publication of a strategy to improve the life chances of disabled people. Of particular concern is the lack of data on impairment types and how impairments interact with social barriers erected by society and its institutions, leading to, PMID- 19562910 TI - Gestation-specific infant mortality by social and biological factors among babies born in Enland and Wales in 2006. PMID- 19562911 TI - Determinants of post-caesarean wound infection at the University College Hospital Ibadan Nigeria. AB - OBJECTIVE: To determine the risk factors that predispose patients to post caesarean wound infection at a tertiary health institution in developing country. METHOD: It was an observational study of all women that had caesarean (elective and emergency) delivery between July and September, 2004. The outcome of their post-caesarean wound was assessed. Statistical analysis (Bivariate and logistic regression) of the identified risk factors in patients who developed wound infection was performed at a 95% level of confidence. RESULTS: The post-caesarean wound infection rate was 16.2%. The identified risk factors were lower educational status, multiple pelvic examination, offensive liquor at surgery and patients that are unbooked. Following logistic regression, women with up to primary school were 20 times more likely than those with secondary education and above to develop wound infection (95% CI OR = 1.8 to 250.0). CONCLUSION: The outcome showed that the identified determinants of post-caesarean wound infection were comparable with earlier reports. Women with lower educational status are more at risk in this study. Therefore, these women need special attention to reduce the risk. PMID- 19562912 TI - Prothrombin time, activated partial thromboplastin time and other haematological parameters among non-diabetic hypertensive patients. AB - OBJECTIVES: To screen non-diabetic hypertensive Nigerian patients with a view to determining their haematological and coagulation profiles. SUBJECTS AND METHODS: Thirty (30) consecutive non-diabetic hypertensive patients, aged 20-60 years and 30 age and sex-matched, apparently healthy subjects, were investigated for some haematological and coagulation parameters, blood pressure, pulse, body weight, height and body mass index. RESULTS: There were statistically significant differences (p < 0.05), in some haematological parameters, blood pressure, body mass index and height in the total number of patients studied when compared with the total control group. The separate genders each showed statistically significant differences in both haematological parameters and blood pressure (P < 0.05) when compared with their corresponding control groups. In addition, haematological values, body mass index and height showed statistically significant differences (p < 0.05) when the male and female patients were compared. Overall, 25 (83.33%) of the patient's blood films were normochromic and normocytic. CONCLUSION: It appears that hypertension has adverse effects on haematological parameters. We therefore recommended that coagulations studies should be included in the investigation work-up of hypertensive patients. PMID- 19562913 TI - Evaluation of commercial HIV test kits used in Nigeria. AB - BACKGROUND: Accurate and reliable diagnosis of HIV plays a central role in any effective HIV intervention. We decided to evaluate 4 commercial HIV test kits to determine their reliability for use in developing countries. METHODS: Serum samples obtained from clients accessing tertiary health services at the STI clinic, Jos University Teaching Hospital were used to evaluate Sdbioline, Diaspot, Determine and DIALAB Elisa kits. A Western blot was used as the reference kit. RESULTS: DETERMINE kit gave 34 positive and 58 negative reactions and the positive sera were all confirmed by Western blot while DIASPOT kit gave 27 false negative results, which was at variance with the reference kit result. Other kits were SDBIOLINE with 5 false positive and DIALAB Elisa kit, which gave one false positive, and one false negative result. CONCLUSION: We conclude that Determine, SDbioline and DIALAB Elisa kits are reliable for HIV antibody testing in Nigeria and other developing countries. PMID- 19562914 TI - Trends in acute emergency room hypertension related deaths: an autopsy study. AB - INTRODUCTION: Hypertension is associated with increased morbidity and mortality. Paradigm shift and novel drugs that go beyond blood pressure control have debuted in the last decade globally and in Nigeria. The study therefore proposed to investigate the effect of the above if any on hypertension related acute deaths in patients admitted to the emergency room of the Lagos University Teaching Hospital. METHOD: Autopsy reports for bodies deposited from the medical emergency room (ER) were reviewed. Details of the time of admission, time of death and blood pressure status prior to the event were obtained. Subjects were batched into two groups 1982 1991 and 1992 2001 based on periods of paradigm shift in hypertension diagnosis and management. RESULT: There were 297 hypertension related deaths but 252 were analyzed. There were 168 (66.7%) males and 84 (33.3%) females (M:F 2:1) and mean age was 47.33 +/- 12.18 years (14-85 yr). Two thirds of the subjects (65.5%) were = 50 yrs of age. The mean duration of admission was 5.88 +/- 6.41 hours. One third (35.3%) died within an hour of admission. The commonest causes of death were stroke (52.8%) and heart failure 103 (40.9%). Intra-cerebral hemorrhage was the commonest type of stroke seen, 69 (52.3%). There were fewer cases of acute deaths in the second decade under review 95 (37.7%) vs. 157 (62.3%), p = 0.02 and strokes during this period, 47 (49.47) Vs 86 (54.78), p = 0.06. CONCLUSION: There is a trend towards reduction of hypertension related acute deaths. However stroke remains a major cause of acute hypertensive death and the patients are still dying young. PMID- 19562915 TI - Biochemical infertility among females attending University of Ilorin Teaching Hospital, Nigeria. AB - BACKGROUND: Biochemical laboratory investigations potentially contribute to the diagnosis of over 50-75% of couples being investigated for infertility. Both hormonal and anti-hormonal treatments have achieved great successes in the treatment of infertility. Our aim therefore was to investigate the pattern of biochemical abnormalities in females diagnosed as infertile form anovulation. MATERIAL AND METHODOLOGY: One hundred and twenty women diagnosed clinically as primary or secondary infertility from anovulation referred from the gynecological clinic of UITH and private hospitals in Ilorin were investigated by routine fertility test profile. RESULT: The age ranged between 20-40 years (mean = 32.9, sd +/- 4.7) for the primary infertility and 23-47 years (mean = 34.4, sd +/- 5.4) for the secondary infertility groups respectively. Ninety six (80%) subjects were found to have hormonal abnormalities. Pattern of biochemical diagnosis amongst the 33 (34.4%) primary infertility subjects included hypergonadotrophic hypogonadism 21 (63.6%), hypogonadotrophic hypogonadism 9 (27.3%), and hyperprolactinemia 3 (9.1%). Among the 63 (65.6%) cases of secondary infertility, there were 31 (49.2%) cases of hypergonadotrophic hypogonadism, 30 (47.6%) hypogonadotrophic hypogonadism, and 2 (3.2%) hyperprolactinemia. There was no statistical difference in the mean values in the various biochemical parameters. CONCLUSION: Hormonal profile should be a goal standard in the diagnosis of anovulation. PMID- 19562916 TI - Computerized tomography of children with seizure disorders. AB - BACKGROUND: Neuro-imaging is generally considered as part of the evaluation of seizures and epilepsy. There is limited information about its usefulness in our environment. We describe the pattern of CT findings in children with seizures in our environment. METHOD: We carried out a retrospective review of the computerized tomography findings in children with recurrent seizures over a one year period, November 2005 to October 2006. RESULTS: During the study period, 49 infants and children had computerized tomography performed on them out of which 19 had CT done for recurrent seizures. They ranged in age from 4 months to 16 years with 13 of them being boys. Generalized tonic--clonic seizures was the most predominant seizure type, being present in 10 of the 19 (52.6%) children while simple partial seizure, myoclonic jerk and mixed seizure types were present in 2 cases each. Abnormal scan was demonstrated in 10 of the 19 children (52.6%) with 3 of them having double cerebral lesions, giving a total of 13 cerebral lesion demonstrated by the CT scan. Cerebral infarct was the most common lesion demonstrated, being present in 5 of the 13 lesions (38.5%). Others were cerebral atrophy in 4 cases (30.8%), moderate ventricular dilatation 2 (15.4%) and 1 each ofporencephalic cyst, hydrocephalus and linear skull fracture. Of the 10 children with abnormal scan, 90% of them had significant past medical history, with birth asphyxia (44.4%) and meningitis (33.3%) being the commonest significant past medical history in them. CCONCLUSION There is a high incidence of abnormal scan findings in children with seizure disorder in our environment compared to what is obtained from the developed countries. Cerebral infarct appears to be the most common abnormal CT findings in our children with seizures. PMID- 19562917 TI - Pre-hospital care in Nigeria: a country without emergency medical services. AB - OBJECTIVES: Efficient pre-hospital transport (emergency medical services, EMS) is associated with improved outcomes in road traffic injuries (RTI). This study aims to discover possible interventions in the existing mode of transport. METHODS: Persons bringing all RTI victims to the Emergency room (ER) over a 4-year period and the injury arrival intervals were noted prospectively. FINDINGS: There were 2,624 patients (1,886 males and 738 females); only 2,046 (78%) had clear documentations of three categories of persons bringing victims to ER: Relatives (REL, 1,081, 52.83%); Police/Federal Road Safety Corps (P/F, 827, 40.42%) and Bystanders (BS, 138, 6.74%). No intervention was provided during transport: Within 1 hour, 986 victims (48.2% of 2,046) arrived ERbrought by P/F (448, 21.9%), REL (439, 21.5% of 2,046), and BS (99, 4.8%). These figures, in each instance, represent 40.6 % of total victims brought by REL; 54.2% by P/F and 71.7% by BS. However, after 6 hours, REL were the main active group as they brought 94.5% (359 of 380) patients of this period. In 91 victims (4.4%) the injury arrival time was not captured. CONCLUSION: This study has identified three groups of persons involved in pre-hospital transport with nearly 50% getting to ER within 1 hour without any intervention or prior notification of ER. Absence of EMS obscures pre-hospital death records. The P/F responsible for only 40% of transport should be trained and equipped to offer basic trauma life support (BTLS). The REL and BS (both responsible for 60% of transport) represent a pool of volunteers for BTLS to be trained. PMID- 19562918 TI - Prevalence of, and attitude towards, needle-stick injuries by Nigerian gynaecological surgeons. AB - Health care workers who have occupational exposure to blood and other potentially infectious materials are at increased risk for acquiring blood-borne infections. The emotional impact of a needle-stick injury can be severe and long lasting, even when a serious infection is not transmitted. OBJECTIVE: To assess the prevalence and attitude towards needle-stick injuries by Nigerian gynaecological surgeons. METHODOLOGY: A cross-sectional study was conducted at the 40th Annual General Meeting and Scientific Conference of the Society of Gynaecology and Obstetrics of Nigeria (SOGON) held in Ibadan, southwest Nigeria from the 23rd to the 26th of November 2005. Data was collected using a self-administered questionnaire. RESULTS: Seventy two questionnaires out ofa hundred administered were finally analysed. Sixty-five (90.3%) respondents had experienced needle stick injuries in the workplace. This occurred in the majority of cases (86.2%) during suturing. Only 9.2% of those experiencing a needle-stick injury took the correct or appropriate action afterwards. Consultants were not significantly more likely than Residents to take appropriate actions after needle-stick injuries (p > 0.10, X2 = 2.11, 1 df). Fifty-two (80%) of those with needle-stick injuries did not report the incident to the appropriate office. Only 26 (37.1%) of 70 respondents indicated the presence of a needle-stick policy in their centres. CONCLUSION: The prevalence of needle-stick injuries among sampled Nigerian gynaecological surgeons is high. Majority are either unaware or do not take appropriate actions after exposure to hazardous body fluids from needle-stick injuries, either through first-aid steps or post-e4posure prophylaxis. All health institutions should have a working needle-stick policy in their centres, and health care workers continually educated on it. PMID- 19562919 TI - Hospital frequency of large bowel cancer: factors thought to influence outcome. AB - BACKGROUND: One of the leading causes of death from malignancies is cancer of the large gut. Elsewhere in the developed world, the disease severity and operative mortality are falling due to increased use of screening, earlier diagnosis and improvement in surgical techniques. AIM: To report our experience with this disease at the Usmanu Danfodiyo University Teaching Hospital (UDUTH), Sokoto. PATIENTS AND METHODS: This is an 8-year retrospective analysis (January 1998 December 2005) of all established cases of large bowel cancer managed at the surgical services of the Usmanu Danfodio University Teaching Hospital (UDUTH), Sokoto. RESULTS: A total of 40 cases of colorectal cancer were seen in this centre during the study period. There were 21 (52.5%) males and 19 (47.5%) females, representing a male: female ratio of approximately 1:1. The modal age was the 5th decade. The youngest patient was 15 years old. Forty-two percent of patients were below 40 years of age. All were symptomatic at time of detection. There was no patient in Dukes' stage A category. Nineteen (47.5%) were stage D at presentation. Potentially curative resection was performed in 5 (12.5%) patients. Another 22 (55.0%) cases had palliative surgical treatment. The remainder were either inoperable (n= 3; 7.5%), refused colostomy (9; 22.5%) or died before surgical treatment (1; 2.5%). Among those who were surgically treated, 6 (22.2%) died within 30 days of operation. The mean duration of post-operative follow up was 16 months (5 67 months). CONCLUSION: Cancers of the large intestine are not uncommon in this part of the world. A rising frequency of colorectal cancers in our locality is observed. Presentation to hospital is usually late. A significant number of patients are below 40 years of age. PMID- 19562920 TI - Evaluation of "care of the foot" as a risk factor for diabetic foot ulceration: the role of internal physicians. AB - BACKGROUND AND OBJECTIVES: Several risk factors predispose the diabetic patient to foot ulceration, including "inadequate care of the foot". This risk factor for foot ulceration has not been previously evaluated among Nigeria diabetic patients and is the objective of this study. SUBJECTS AND METHODS: One hundred and twenty (120) diabetic patients with and without symptoms of peripheral neuropathy receiving care at the medical outpatient department (MOPD) and the diabetic clinic of the Nnamdi Azikiwe University Teaching Hospital Nnewi were recruited consecutively as they presented. They were administered structured questionnaires to assess some variables concerning care of their feet as provided to them by their physicians. RESULTS: Among the 120 diabetic participants, 83 (69.2%) had neuropathic symptoms (the symptomatic participants) while 37 (30.8%) were asymptomatic (the asymptomatic participants). Eighty (80; 96.4%) of the symptomatic vs 36 (97.3%) of the asymptomatic participants had never had their feet examined by their physician. Also, 26 (31.3%) of the symptomatic vs 12 (32.4%) of the asymptomatic participants had never received any form of advice on how to take special care of their feet by their physician, and 26 (31.3%) of the symptomatic vs 6 (16.2%) of the asymptomatic participants walked unshod most times in their immediate surroundings. CONCLUSION: Physicians do not provide adequate care to the feet of their diabetic patients irrespective of the presence or absence of neuropathic symptoms, making this variable a critical risk factor for diabetic foot ulceration and amputation. Continuing medical education to health care providers emphasizing adequate "care of the foot" of the diabetic patient, will reduce avoidable loss of limbs to diabetes. PMID- 19562921 TI - Pattern of gynaecological consultations at Ladoke Akintola University of Technology Teaching Hospital. AB - AIMS: A descriptive study to determine the pattern of gynaecological consultations at Ladoke Akintola University of Technology Teaching Hospital, and to know the factors that determine the pattern as well as the outcome of such consultations. MATERIAL AND METHODS: The case notes of all the patients that attended the gynaecology clinic between 1st January 2004 and 31st December 2005 were studied. RESULTS: Two hundred and fifty patients attended the gynaecology clinic during the study period, however, 208 (83.2%) case notes were available for analysis. Most of the patients that had gynaecological consultations during the study period were married (88.0%), and infertility (48.1%) was the commonest reason for the consultations. Many of the patients (60.1%) did not receive definitive treatment for their consultation. CONCLUSION: Infertility being the commonest reason for gynaecological consultations, and tubal factor the commonest cause of infertility, there is need to establish assisted reproductive technology in public health institutions to render services at a highly subsidized rate in helping this population of patients who could not afford this treatment which is presently being offered by private health institutions. PMID- 19562922 TI - Day case haemorrhoidectomy in a developing country. AB - BACKGROUND: Ligation excision haemorrhoidectomies are usually done on inpatient basis. Over the years however, there has been an increase in the numbers done on outpatient basis. This retrospective review was conducted to evaluate the results of day case haemorrhoidectomy in a developing country. MATERIALS AND METHODS: Forty three consecutive patients, who met the criteria for day case surgery, had ligation excision haemorrhoidectomy, between January 2004 and September 2005 at the Royal Victoria Teaching Hospital (RVTH) Banjul with the intention of same-day discharge from hospital. For each patient, data collected included age, sex, duration of symptoms, degree of haemorrhoids, and duration of rest after the procedure in minutes or hours, whether the patient was discharged same day or needed admission for complications. Local anaesthesia, 1% lignocaine was used for all the patients. RESULTS: Forty three patients comprising 28 males (65.1%) and 15 females (34.9%), with a male female ratio of 1.87:1 underwent planned day case haemorrhoidectomy. Their mean age was 36.27 +/- 10.26 years and range was 25 to 56 years. Forty patients (93%) were discharged on the same day of surgery after a rest period in the day care theatre ranging between 25 minutes and 60 minutes with a mean of 41.2 +/- 11.08 minutes. Three (7%) of the patients required admission, two for acute urinary retention and control of excessive pain and the other 5 days postoperative on account of secondary haemorrhage. Pain control was achieved by oral non steroidal antiinflammatory agents. There were no deaths and majority of the patients were satisfied with the procedure. CONCLUSION: Ligation excision (Milligan-Morgan) haemorrhoidectomy is safe and can be performed successfully on out patient basis, saving inpatient health care costs. This is suitable for developing country. PMID- 19562923 TI - Time of passage of first stools (meconium) and serum levels of calcium and magnesium in Nigerian neonates (African neonates). AB - BACKGROUND: Over 90% full term normal newborns are known to pass meconium (first stools) within the first 24 hrs of life especially among Caucasians. This has not been fully documented among African neonates. This study was therefore done to determine time of passage of first stools (meconium) in African neonates and also the serum levels of calcium and magnesium in these children in order to establish a data base for normal standards. METHODOLOGY: Two thousand and four hundred neonates were recruited from Hospitals in Benin City urban areas for the study. 76.7% of these were full term and AGA babies (37-40 weeks; 2.8 kg-4.3 kg) while 23.3% of them were preterm and low birth weight neonates (34-36 weeks and weight between 2.4 to 1.5 kg) respectively. RESULTS: 88% of the full term neonates passed meconium at 15.4 +/- 3.6 hrs of life while only 12.0% of them passed meconium after 24 hrs of life and all by 48 hrs of life. The mean time of passage of stools by preterm, low birth weight babies was 45.2 +/- 2.4 hrs. The values of serum calcium in the full term neonates ranged from 6.5 9.2 mEq/l with mean of 7.8 +/- 1.2 mEq/l while the level in the preterm low birthweight neonates was much lower (5.4 8.3 mEq/l) with a mean of 6.7 +/- 1.3 mEq/l. The values of serum magnesium ranged from 0.9 1.6 mEq/l with a mean of 1.2 +/- 0.3 mEq/l in full term neonates while preterm low birthweight neonates had levels as low as 0.5 1.1 mEq/l with a mean of 0.7 +/- 0.2 mEq/l CONCLUSION: This study has shown a relatively shorter time of passage of first stools (meconium) in Nigerian neonates (blacks) than in the Caucasians. The implication of the findings in this study is that a delay in the passage of first stools (meconium) and early appearance of jaundice in normal black neonates could be due to gastrointestinal abnormalities. This observation could lead to early identification of these neonates with resultant early intervention. PMID- 19562924 TI - Haematological assessment of occupational exposure to lead handlers in Enugu urban, Enugu State, Nigeria. AB - OBJECTIVE: To determine blood levels of lead and its effects on haematological parameters among occupational lead handlers in Enugu, Enugu State, Nigeria. In developing countries, rapid industrialisation has lead to an alarming demand for lead. Furthermore, the burden of lead toxicity is greatly underestimated. Hence, the need to assess the unavoidable toxic effects of lead as done in this study. METHODS: Blood lead levels were measured by atomic absorption spectrometry (AAS) in eighty one (81) male subjects from three manufacturing companies, all located in Enugu metropolis, Nigeria. Thirty (30) staff of the industries not directly involved in lead handling served as control group I, while twenty (20) apparently healthy individuals from within the same locality not involved in lead handling served as control group II. Haematological values, blood lead levels and blood pressure (BP) were established using standard procedures. Statistical Analysis System (SAS) software was used to analyze the results. P value of < 0.05 was taken as significant. RESULTS: Mean blood levels were 7.00 +/- 0.07 microg/dl in test subjects; 3.00 +/- 0.19 microg/dl in control group I and 2.00 +/- 0.04 microg/dl in the control group II. There were significant statistical differences (p < 0.05 for each) in haemoglobin (Hb), packed cell volume (PCV), reticulocyte, total white blood cell (WBC), monocyte, autohaemolysis without glucose, and systolic and diastolic pressure between subjects and control group I. There were also significant differences (p < 0.05 for each) in the mean levels of Hb, PCV, reticulocyte, eosinophil, monocytes and systolic and diastolic pressures between the test subjects and control group II. There were however, no statistically significant differences (p > 0.05) in the means of other parameters. Basophilic stipplings were not observed in the red cells of those directly exposed to lead. CONCLUSIONS: It is suggested, therefore, that comprehensive and preventive measures towards exposure to lead in work places, and routine haemotological investigations be included in the bio-monitoring of the health status of lead workers. PMID- 19562925 TI - Seroprevalence of Hepatitis C viral antibodies in pregnancy in a tertiary health facility in Nigeria. AB - BACKGROUND: Liver disease due to Hepatitis C viral (HCV) infection is the most common indication for liver transplant. It is a viral pandemic that is five times as widespread as the human immunodeficiency virus type 1 infection. In spite of this, vaccines were yet unavailable for protection of the human race due to the morphology and fastidious nature of the organism. While the scanty data available on this infection in our environment are limited to blood donors, people continue to be screened for and deprived of renal dialysis if any patient is found to have HCV infection. Also in this environment, data on HCV infection in pregnancy is virtually nonexistent even though the infection can have a deleterious effect on materno-fetal outcome. OBJECTIVE OF THE STUDY: To determine the seroprevalence of hepatitis C viral antibodies among antenatal women attending a tertiary health facility in Nigeria. METHODOLOGY: This was a prospective cross-sectional study whose subjects were booked consecutive antenatal women volunteers attending the University of Benin Teaching Hospital, Benin City, Nigeria between June 1 and December 31, 2005. Hepatitis C viral antibodies were determined and confirmed using a second and a third generation Enzyme Linked immunosorbent assay respectively. Both HCV sero-positive and seronegative women had both pre-and post test counseling. RESULTS: Of the 269 samples screened for HCV antibodies, 5 (1.86%) samples were confirmed seropositive. None of the HCV seropositive women had liver enzyme derangement. CONCLUSION: Hepatitis C viral infection in pregnancy is not uncommon in Nigeria. It's prevalence in pregnant women South South of Nigerian is similar to that of their Cameroonian counterparts, an immediate neighbouring country. A multi-centre study to determine the national prevalence of HCV and in addition to elevation of public awareness is suggested. Hepatitis C viral-induced liver disease remains the major indication for liver transplant for which our present levels of economy and health infrastructures can least support. With no vaccines and no cure, the time to act is now. PMID- 19562926 TI - The functional status of disease surveillance and notification system at the local government level in Yobe State, Nigeria. AB - BACKGROUND: An effective Disease surveillance system provides information that is required for appropriate action. Although evaluated by attributes like sensitivity, predictive value and representativeness, the assessment of its operational status is critical, especially in developing countries. METHODS: We conducted this cross sectional survey among 144 health personnel from facilities and Local Government disease surveillance officers, using pre-tested, semi structured, self-administered questionnaire and observational checklist. RESULTS: This showed that only 55 (38.2%) of the respondents were aware of the Disease surveillance and notification (DSN) system. Fifty-eight (65.9%) and 7 (8.0%) of the facilities had up-to-date registers and DSN forms respectively. Diagnostic support was lacking in most of the health facilities. Data was not analysed at LGA level, and forms and logistics for supervising disease surveillance activities and feedback were inadequate. CONCLUSION: We recommend training/retraining of health personnel on infectious disease surveillance, provision of logistics, improved supervision and feedback of information. PMID- 19562927 TI - Changing pattern of antibiotic sensitivity of Neisseriae meningitidis from children with Meningococcal meningitis in North Eastern Nigeria. AB - OBJECTIVE: To determine the changing pattern of antibiotic sensitivity of Neisseriae meningitidis from children with Meningococcal meningitis in North Eastern Nigeria. MATERIALS AND METHODS: This study was carried out over 31 months, from January 2003 to July 2005 in the Paediatric ward of Federal Medical Centre (FMC), Azare. The cerebrospinal fluid (CSF) of all patients with clinical features of meningitis admitted were examined microbiologically, including culture for bacterial organisms and their sensitivity pattern as well as biochemical tests determined. RESULTS: The CSF specimens from 44 patients, aged between 2 months and 12 years (20 females and 24 males) yielded gram negative intracellular diplococci by gram staining. Of these, Neisseriae meningitidis was cultured in 18 (40.9%). The remaining bacterial isolates did not survive culture. The analysis of antibiotic sensitivity of Neisseriae meningitides from the eighteen CSF specimens showed 100 per cent sensitivity to ceftriaxone. Resistance of the bacterial isolates to benzyl penicillin, ampicillin and chloramphenicol was 80%, 71.4% and 20% respectively. Varying levels of sensitivity of the organisms to augmentin (amoxicillin-clavulanic acid), gentamicin and cotrimoxazole were also documented. CONCLUSION: This study highlights the increasing resistance of Neisseriae meningitidis to empirically used antimicrobial drugs. PMID- 19562928 TI - Evaluation of serum uric acid levels in normal pregnant Nigerian women. AB - BACKGROUND: Hypertensive disorders in pregnancy are common in our environment. The aetiology is unknown and the prognostic indicators of the severity of maternal and fetal complications are variable. The level of uric acid, which is one of the prognostic indicators, is altered in normal pregnancy and as pregnancy advances. Base line values are thus extremely important to enable reasonable prognostic assessment in hypertensive pregnancies. OBJECTIVES: To determine levels of serum uric acid during normal pregnancy in University of Nigeria Teaching Hospital (UNTH) Enugu. STUDY DESIGN: settings and methods: Sixty- five pregnant and 65 non-pregnant women with age range 20-38 years were recruited. The pregnant women were in their second and third trimesters, attending antenatal clinic at the University of Nigeria Teaching Hospital Enugu. Serum levels of uric acid were determined for the entire subjects. RESULTS: The serum uric acid levels were significantly lower in the pregnant women than in controls (P < 0.001). 0.15 +/- 0.03 mmol/L in the second trimester, 0.14 +/- 0.02 mmol/L in the third trimester and 0.29 +/- 0.04 mmoL for control. CONCLUSION: The low levels in pregnancy and as pregnancy progresses should be taken into consideration when monitoring hypertensive disorders in pregnancy using serum uric acid. Thus levels that are within normal for non pregnant population may indeed be an indication for intervention in pregnancies complicated by preeclampsia. PMID- 19562929 TI - Glycated haemoglobin and glycaemic control of diabetics in Ilorin. AB - OBJECTIVES: With increasing adoption of Western Lifestyle in Nigeria, the incidence of Diabetes Mellitus is on the increase in the country with its attendant complications. The objective of this study was to determine the prevalence of patients at risk of developing diabetic complications in Ilorin, Nigeria, among our patients with diabetes mellitus. METHODS: A cross-sectional study of diabetic patients attending the University of Ilorin Teaching Hospital ted haemoglobin as an index of medium term glucose control was assayed in established diabetics. The result obtained was evaluated against the bench mark HbA1c value of 7.2% for the development of complication. RESULTS: Fifty-six percent of the subjects were females and all of them were forty years and above in age. Only four percent of the patients were below the age of forty years. Seventy-two percent of the subjects had diabetes for less than 10 years. Only female patients had BMI values greater than 30 kg/m2. About 64% of the patients had HbA1c value > 7.2%. More males (73.7%) had HbA1c 7.2% than females (64.5%) (P < 0.05). The patients had a mean HbA1c value of 8.0%, while the mean HbA1c in the control was 5.2%. These two mean HbA1c values gave a P-value of 0.0001 on Student t-test. The female diabetic patients had a mean HbA1c value of 7.8% (SD = 1.96) against the value of 5.1% (SD = 1.13) for the female control patients (P-value of 0.0001). Similarly, the male patients and male control subjects had mean HbA1c values of 8.1% (SD = 1.96) and 5.6% (SD = 1.00) respectively with P-value of 0.0001. The control subjects had a mean fasting blood glucose level of (+/- SD) 4.93 +/- 1.09 mmol/L and the corresponding value for the diabetics was 8.5 +/- 4.2 mmol/L. when these two values were compared we got a P-Value < 0.05. CONCLUSIONS: The mean HbA1c values between the patients and the control subjects were significantly different. Diabetics in our environment with mean HbA1c value of 8.0% are prone to developing complications because of poor glycaemic control. We therefore advise that, periodic estimation ofglycated haemoglobin be carried out along side fasting blood glucose, in our diabetics. PMID- 19562930 TI - Ethical considerations in women's sexual and reproductive health care. AB - BACKGROUND: The concept of reproductive health, which adopts a holistic approach to the management of reproductive disorders, was developed in 1994 at the International Conference on Population and Development. This together with the adoption of a right-based approach to the relationship between reproductive health and population and development and the emergence of the concept of sexual and reproductive right marked a turning point in contemporary global health care initiative. Sexual and reproductive healthcare raises ethical questions that fall within the purview of bio-ethics. OBJECTIVE: To review ethical issues related to women's sexual and reproductive healthcare. METHODOLOGY: A critical review of available literature on the subject matter was conducted. FINDINGS: Four key principles form the basis of Bioethical analysis--respect for person, beneficence, non-maleficence, and justice, applicable at four different levels microethical, macroethical, mesoethical, and megaethical levels each of which can be employed in the ethical analysis of sexual and reproductive health care. Medical practitioners caring for women, for the fact that they work in areas of a woman's body that are of particular psychosocial sensitivity, are expected to adhere to strict ethical principles in their practice. The International Federation of Gynaecology and Obstetrics (FIGO) in collaboration with the Society of Gynaecology and Obstetrics of Nigeria (SOGON) has developed a human right based code of ethics related to sexual and reproductive health care to guide medical practitioners caring for women in their daily practice. CONCLUSION: An understanding and proper application of the ethical principles is expected to enable these medical practitioners to actualize the ultimate and desired goal of uplifting the sexual and reproductive healthcare and right of women. PMID- 19562931 TI - Inevitable myomectomy during caesarean section: a case report. AB - Myomectomy during Caesarean section is conventionally reserved for pedunculated myomas. When fibroids located in the lower uterine segment are encountered in pregnancy, due to fear of haemorrhage, obstetricians would prefer to perform a classical Caesarean section even with its high risk of subsequent uterine rupture and avoid the fibroids. We hereby present a case where removal of intra-mural fibroids in a 34-year old primigravida though inevitably performed, was successful. PMID- 19562932 TI - Squamous cell carcinoma of the conjunctiva in Sagamu, Nigeria--a review and 2 case reports. AB - AIM: To review the current literature on squamous cell carcinoma [SCC] of the conjunctiva and report 2 presentations in Sagamu, Nigeria since the availability of histopathological services. METHODS: The available literature using e-medicine and pubmed internet websites were summarized. The 2 cases seen by the first author since histopathological services started at Olabisi Onabanjo University Teaching Hospital [O.O.U.T.H.], Sagamu were summarized using the case note records and the pathological reports of their surgical specimens. RESULTS: The 2 cases showed the variable ways SCC of the conjunctiva can present in an equatorial African country like Nigeria. DISCUSSION: Limbal interpalpebral conjunctival lesions should arouse the possibility of this rare neoplasia even in patients with no evidence of predisposing factors. CONCLUSION: Even though SCC of the conjunctiva is commoner in Caucasians, its possible presentations should be kept in mind in equatorial non-Caucasian populations like Nigeria. PMID- 19562933 TI - Chronic uterine inversion secondary to submucous fibroid: a case report. AB - A rare case of non-puerperal chronic uterine inversion secondary to sub mucous fibroid in a 38-year-old woman is presented. There was complete uterine inversion with the incarcerated inverted uterus protruding through the vvgina beyond the vulva. The sub mucous fibroid was attached to the fundus. At laparotomy, a dimple with a constriction ring was found in the position of the uterus. The distal ends of the fallopian tubes and part of the ovary were visible through the constriction ring. Histological examination of the uterus and fibroid following hysterectomy confirmed their benign nature. PMID- 19562934 TI - Concurrent use of cyclophosphamide and prednisolone in childhood nephrotic syndrome in South-East Nigeria; a report of 5 cases. AB - INTRODUCTION: Nephrotic syndrome is a chronic renal disease that can lead to end stage renal disease. There are different histological types with global variations in frequency. Literatures reviewed showed that the African variant is less likely to be minimal change variant. No clear treatment protocol for it has been most beneficial. OBJECTIVE: This paper aims to evaluate the outcome of a treatment protocol using cyclophosphamide and prednisolone concurrently. METHODS: A low dose and short duration concurrent use of cyclophosphamide and prednisolone was used for treating children with nephrotic syndrome who had not developed derangements of their renal function. The case files of those that were treated and followed up over a 10 year period or until they were above 18 years of age were analysed for their clinical parameters. RESULTS: Five cases were treated and all have been in clinical remission for more than 4 years as at the time of the review, though 2 of them relapsed twice initially. They were all aged above 6 years and had microscopic hematuria. The 3 cases whose ESRs were done had high levels. Two cases that presented 1 yr after the onset of their symptoms resolved without relapse while 2 out of the remaining 3 in whom this interval was less than 6 months relapsed. CONCLUSION: This treatment protocol appears beneficial to childhood nephrotics in this environment and should be used. PMID- 19562935 TI - [Cell cycle regulation by F-box protein Fbxw7]. PMID- 19562936 TI - [Revisiting sensitization mechanisms in cancer thermochemotherapy]. PMID- 19562937 TI - Case of critical complication after surgery for ankylosing spinal hyperostosis. AB - We experienced a case who received a bone resection for ankylosing spinal hyperostosis through the anterior approach. He subsequently became asphyxic and suffered a cardiopulmonary arrest owing to a postoperative hematoma. His complaint before the surgery was sticking of his throat and dyspnea that continued for 3 years. X-ray films revealed an osteophyte on the anterior side of the C3-C6 vertebral bodies, and we made an anterior approach under general anesthesia. There were no problems during the surgery. After being returned to the ward without intubation, he complained that he had a catch in his throat that progressed little by little, and subsequently suffered a cardiopulmonary arrest at 5 hours after surgery. Upon revival, he had brain hypoxia. He passed away owing to malnutrition and pneumonia at 4 years after the surgery. To prevent this complication, it is important to have an understanding of this condition. We should have alerted the nurses that such a complication may occur after anterior spinal surgery. It is also important to be aware that intubation of such a case becomes difficult once the trachea has become compressed and curved because of a hematoma. PMID- 19562938 TI - [Functional model of the middle ear ossicles]. AB - In students' dissection practice, it is very difficult to teach students the structures and functions of the middle ear ossicles. The middle ear ossicles are too small to explain their structures and functions. Models are useful in explaining these points, but there have been no models that accurately explain the movements of the middle ear ossicles and the functions of the muscles in the middle ear. This time, we have made a model of middle ear ossicles. Our ear ossicles are made of paper-mache with metal in it. The incudomalleolar and incudostapedial articulations are made of rubber. The tensor tympani and the stapedius muscles are made of wire and the two wires can be fixed by cord stoppers. Our model explains clearly the following mechanisms of the middle ear ossicles. 1. The mechanism of sound conduction system. When the sound vibrates the tympanic membrane, malleus and incus rotate together. The long process of the incus pushes the head of the stapes. The sound is amplified by leverage. 2. Attenuation of sound by contractions of tensor tympani and stapedius muscles. When a loud sound is transmitted through the ossicular system, the tensor tympani muscle pulls the malleus inward while the stapedius muscle pulls the stapes outward. These two forces oppose each other and increase rigidity of the ossicular system, thus reducing the ossicular conduction. 3. The mechanism of how paralysis of stapedius muscle, caused by an injury to the facial nerve, results in hyperacusis. 4. This model also suggests a possible reason why the pars lucida of the tympanic membrane exists. PMID- 19562939 TI - [What clinicians expect from animal models of depression]. AB - In depression research, animal models are essential for better understanding of biological mechanisms, development of new agents, and showing evidence on various theories. Given the emotional feature of the disorder, people have debated, even revolted, on applying animal models to human studies. However, this logic is counter-arguable by noting the commonality of biological changes (e.g., behavioral disorder, environment maladjustment, and stress vulnerability) between human and experimental animals; moreover, genetic studies suggest further similarities. However, current animal models are insufficient to express the full dimension of mood disorders. The model concepts are often in a narrow context of antidepressant responses. The models consider stress models as equivalents of depression models. This hypothesis, however, has limitations in differentiating anxiety- and stress-related models from depression models. The models tend to focus on the depressive, but not manic, state; it is thereby a challenge to replicate depressive and manic phases in animals. The models do not have a standpoint of depressive disorder from cognitive aspects, and are unable to replicate mood disorder subtypes, such as atypical depression, dysthymia, bipolar disorder, and seasonal affective disorder. From a clinical perspective, models of treatment-resistant depression would eventually be of large benefit to enhance treatment outcomes in clinical settings. PMID- 19562940 TI - [Validity of forced swimming as an animal model of depression]. AB - Forced swimming is a brief and excellent assessment method for antidepressant drugs. However, a complete animal model of depression does not exist, because (1) we can not make sure the existence of psychiatric symptoms by interview, (2) individual clinical symptoms of depression were not disease-specific, and (3) there are few reliable biological markers of depression. We tried to summarize these issues by evaluating the validity of forced swimming as an animal model of depression. PMID- 19562941 TI - [Development of animal models for schizophrenia based on clinical evidence: expectation for psychiatrists]. AB - Schizophrenic patients show positive symptoms, negative symptoms and cognitive dysfunction. In humans, phencyclidine (PCP), a noncompetitive N-methyl-D aspartate (NMDA) receptor antagonist, reproduces the schizophrenia-like psychosis including positive symptoms, negative symptoms and cognitive dysfunction. Ketamine, another non-competitive NMDA receptor antagonist, also reproduces a schizophrenia-like psychosis in healthy volunteers, and exaggerates the psychosis in schizophrenic patients. It has been hypothesized that insufficient glutamate neurotransmission is involved in the pathophysiology of schizophrenia. Therefore, attempts have been made to develop animal models of schizophrenia by using NMDA receptor antagonists such as PCP, ketamine and dizocilpine. In addition to pharmacological approaches, genetic approaches have been adopted to develop animal models of schizophrenia. The disrupted in schizophrenia 1 (DISC1) gene has been identified as a schizophrenia susceptibility gene based on linkage and single nucleotide polymorphism (SNP) association studies and clinical data, demonstrating that risk SNPs impact on the hippocampal structure and function in clinical and functional roles of DISC1 are analyzed in many kinds of transgenic mice developed. In this review, we focused on PCP and DISC1 transgenic animal models of schizophrenia and summarized recent evidence from several investigators. The basic researchers would need to collaborate with clinical psychiatrists to develop appropriate animal models for schizophrenia based on clinical evidence. PMID- 19562942 TI - [Significance of imaging biomarkers in exploratory clinical trials]. AB - Brain imaging tools such as positron emission tomography (PET) and functional magnetic resonance imaging have been successfully used to investigate in vivo human brain functions. In particular, PET has been widely used to visualize various molecules such as receptors, transporters, and amyloid proteins. Despite the fact that a considerable amount of time and money is invested in drug development, only a few drugs have been able to complete clinical trials; this can be partly attributed to the lack of appropriate information regarding the in vivo pharmacokinetics and pharmacodynamics of the drugs. Therefore, currently, PET is being employed to determine the optimal clinical dose and the pharmacokinetic profile of psychotropics by investigating the index of drug occupancy at its action sites. Furthermore, recently, the European Agency for the Evaluation of Medicinal Products and Food and Drug Administration have introduced microdose clinical trials and exploratory investigational new drug studies to screen candidate drugs using PET in the early phase of clinical trials. Subsequently, in 2008, the Japanese Ministry of Health, Labor and Welfare also released certain guidelines for microdose clinical trials. Thus, imaging biomarkers have facilitated the collection of useful information on drugs at every stage of drug development. PMID- 19562943 TI - [Clinical evaluation of CNS drugs with PET]. AB - The utility and possibility of PET in the drug development of the neuropsychiatry field is discussed in this review. Positron emission tomography (PET) is a nuclear medicine imaging technique which visualizes and quantifies the distributions of various molecular targets in vivo. This technique has been expected to help the process to be more efficient in the drug development of the neuropsychiatry field. Three approaches have been raised as promising ways: microdosing, occupancy measurement at the target region, and biomarkers. In microdosing, it could be detected whether the drugs enter the living human brain or not using very small amount of radiolabelled candidate drugs. With occupancy measurement, the relationship between the dose of the drug and the occupancy could be established, and the optimal clinical doses could be obtained. As biomarkers, the change in the distribution in beta amyloid and microglia after the drug treatment could be detected. PMID- 19562944 TI - [Strategy for development of imaging biomarkers]. AB - Imaging biomarkers are getting to be more and more important to measure physiological and biological functions, to evaluate higher brain function, to study the pathophysiology of various disorders and to develop effective therapeutic drugs. Positron emission tomography (PET) and single-photon emission computed tomography are representative molecular imaging methods. Development of radiopharmaceuticals is a key for successful in vivo imaging. So far many radioligands have been developed for PET and applied to clinical studies. Development of radioligands has the following process: 1) selection of biomarkers, 2) selection of leading compounds, radionuclide, labeled position, and synthesis methods, 3) in vitro and in vivo evaluation including probability for imaging, selectivity, specificity, and species differences, and 4) evaluation of safety such as acute toxicity, mutagenicity and radiation dosimetry. Clinically, first PET studies were performed in normal subjects, and then recently, radiation dosimetry has been measured directly by dynamic whole body scans. Second PET studies are applied to patients with various disorders. In this manuscript, the author introduces experiences and concepts for development of radioligands for studying the central nervous system. PMID- 19562945 TI - [Evaluation of imaging biomarker by transgenic mouse models]. AB - The invention of trangenic and gene knockout mice contributes to the understanding of various brain functions. With the previous-generation positron emission tomography (PET) camera it was impossible to visualize the mouse brain functions, while the newly developed small-animal PET camera with higher resolution is enough to visualize the mouse brain functions. In the present study, we investigated the visualization of functional brain images for a few transgenic mouse models using the small-animal PET. In neurodegenerative illnesses such as Alzheimer disease (AD), the relationship between etiopathology and main symptoms has been elucidated relatively well; therefore several transgenic mice have been already developed. We succeeded in visualizing amyloid images in human mutant amyloid precursor protein (APP) transgenic mice brains. This result suggested that small-animal PET enabled the quantitative analysis of pathologies in the Tg mouse brain. Psychiatric disorders are presumed to have underlying multiple neural dysfunctions. Despite some efficient medicinal therapies having been already established, the etiopathology of mental illness and its biological markers have not been clarified. Thus, we investigated in type II Ca-calmodulin-dependent protein kinase alpha (CaMKII alpha) heterozygous knockout (hKO) mouse, a major protein kinase in the brain. The CaMKII alpha hKO mice have several abnormal behavioral phenotypes, such as hyper aggression and lack of anxiogenic responses; therefore CaMKII alpha might involve in the pathogenesis of mood disorder and affect personal characterizations. Furthermore, serotonin (5-HT) 1A receptor density in the CaMKII alpha hKO mouse brain changed among various brain regions compared to wild mice. These mechanistic insights, PET assays of Tg mice that we have established here, provide an efficient methodology for preclinical evaluation of emerging diagnostic and therapeutic agents for neurodegenerative and psychiatric illnesses. PMID- 19562946 TI - Delayed sympathetic hyperactivity following electroconvulsive therapy in patients with catatonic schizophrenia. AB - Recently, we encountered patients with catatonic schizophrenia who developed severe tachycardia, atrial fibrillation, and hypertension approximately 10 min after electroconvulsive therapy (ECT). Therefore, to examine whether delayed sympathetic hyperactivity occurs following ECT in patients with catatonic schizophrenia, we performed spectral analysis of heart rate variability (HRV). Nine patients with catatonic schizophrenia, 5 with noncatatonic schizophrenia, and 24 with mood disorders who received ECT consecutively were enrolled. The HRV frequency components were measured at baseline and during each 5-min time interval from the end of the ictal response to 35 min. The power spectrum of HRV was divided into 2 components: a high-frequency component (HF) and a low frequency component (LF). The ratio of LF to HF (LF/HF) is an index of sympathetic activity. LF/HF demonstrated a transient increase between 5 and 10 min after ECT in the catatonic schizophrenia group compared to that in the mood disorder group. ECT in patients with catatonic schizophrenia is associated with delayed, transient sympathetic hyperactivity. These patients may be at an increased risk for developing tachycardia, atrial fibrillation, and hypertension following ECT. PMID- 19562947 TI - [Possible relation of BDNF and GDNF to neuropsychiatric disorders]. AB - Structural abnormalities are demonstrated in various neuropsychiatric disorders, including Alzheimer's disease, Parkinson's disease, and even major depression. On the other hand, recent studies have demonstrated the structural and functional modifications in the adult brain that are associated with synaptic plasticity and neurogenesis. Accordingly, regulation of synaptic plasticity and neurogenesis may lead to the development of novel treatments for neuropsychiatric disorders. Brain derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) have important roles not only in neuronal survival and differentiation, but also in the formation and maintenance of neural circuits and synapse plasticity. Accumulating evidence suggests that these neurotrophic factors may be applied to the treatment of neuropsychiatric disorders. In addition, compounds that increase the expression of BDNF and/or GDNF in the brain should have potential therapeutic values. We have demonstrated that systemic administration of dipeptide Leu-Ile increases BDNF and GDNF production in the brain, and has a protective role in methamphetamine and morphine dependence. In this review, we discuss the potential role of BDNF, GDNF and their inducers in the treatment for neuropsychiatric disorders. PMID- 19562948 TI - The interrogative imperative: state, community, and individual as bodies and subjectivities. PMID- 19562949 TI - Maurice Merleau-Ponty and the "embodied subjectivity" (1908-61). PMID- 19562950 TI - Negotiating community engagement and science in the federal environmental public health sector. AB - In this case study, I use ethnographic data to explore how community engagement and science are deployed at the U.S. Agency for Toxic Substances and Disease Registry, with the goal of formulating an understanding of the personalized meanings of science-community relations for key environmental public health experts. In focus is the cultural discourse circulating in the agency that exposes the real concerns, beliefs, and attitudes of these scientists and experts vis-&-vis their community engagement experiences. Finally, I propose that critical attention to the place of power relations, knowledge politics, and environmental justice are fundamental to studies of toxic contamination where commitments to community engagement and quality science are joined to form a positive research goal and where attempts are made to improve the conditions of quality environmental public health service. PMID- 19562951 TI - Community engagement and science: prospects and provocations: critical commentary on Peter C. Little's "negotiating community engagement and science in the federal environmental public health sector". PMID- 19562952 TI - "You look, thank God, quite good on the outside": imitating the ideal self in a Jewish ultra-Orthodox rehabilitation site. AB - Rather than viewing therapeutic interventions as either compliance or resistance to the social order, I analyze them as mimesis of cultural ideal selves. In particular, I examine the new mediations of the social order constituted in mimetic therapeutic practices and their entailed creativity and ambivalence. Drawing on participant observation in a Jewish ultra-Orthodox (Haredi) rehabilitation site I explore how, given the ruptures brought about in mental disorders, caretakers offer their clients new ways to inhabit the normal self through its imitation. Specifically, caregivers construct replications of dominant selves by selectively deploying modern and neotraditional discourses of the self in diverse social contexts and in multiple registries like body, emotions, social relations, and ways of belonging to the community. I suggest that mimetic therapeutic work is carried out along emerging social distinctions that are associated with Haredim's complex relations with the secular society, and with cultural contestations within the community itself. PMID- 19562953 TI - On becoming a male sex worker in Mysore: sexual subjectivity, "empowerment," and community-based HIV prevention research. AB - Growing public health attention has been placed on the HIV vulnerability of males who sell sex to males in India. However, there is little research that outlines the trajectories through which males come to be involved in practicing sex work in India. Locating "male sex work" within a vibrant social, political, and erotic landscape, this article explores the intertwining of "sexual subjectivity" and "sex work." The authors refer to 70 sexual life histories generated from research conducted in Mysore to unsettle dominant public health notions that regard male sex work as rooted solely in poverty or as a decontexualized "behavioral risk factor." Such perspectives are countered by demonstrating how male sex work in Mysore encompasses a complex interplay between self-realization, sexual desire, social interaction, and public health discourse. Local conceptualizations of selfhood are discussed to suggest the limitations of prevailing empowerment discourses that advance Western notions of individuality. PMID- 19562954 TI - Matters of "conscience": the politics of reproductive healthcare in Poland. AB - The fall of state socialism in Poland in 1989 constituted a critical moment that redefined policies regulating reproductive health and access to care. As the Polish state adopted the discourse and agenda of the Catholic Church in its health policies, reproduction and sexuality became sites of moral governance through the implementation of the Conscience Clause law, which permits healthcare providers to deny medical services citing conscience-based objections. Based on ethnographic fieldwork, this article explores the effects of the implementation of the conscience clause and argues that the adoption of this law for individual use paved the way for restrictions on reproductive healthcare on a systemic scale. The special status afforded to the church is highly significant for access to health services deemed by the church to be matters of morality. The Polish case raises concerns about the place of women's rights in postsocialism and the nature of Polish democratization. PMID- 19562955 TI - The executive summary of the National Forum on the Future of Defense Health Information Systems. AB - The Department of Defense (DoD) has been engaged in the development and deployment of the longitudinal health record (LHR). It has achieved remarkable technological success by handling vast amounts of patient data coming from clinical sites around the globe. Interoperability between DoD and VA has improved and this information sharing capability is expected to continue to expand as the defense health information system becomes an integral part of the national network. On the other hand, significant challenges remain in terms of user acceptance, ability to incorporate innovations, software acquisition methodology, and overall systems architecture. PMID- 19562956 TI - Toward a DoD/VA longitudinal health record: politics and the policy landscape. AB - Policy implications of an interoperable Department of Defense (DoD) and Veterans Administration (VA) longitudinal health record (LHR) are substantial and far reaching. In this manuscript the authors explore the existing challenges and opportunities, the political landscape, and alternative solutions that have created a favorable environment for legislation and funding to support its development. The authors identify six policy themes emerging from the historic National Forum on the Future of the Defense Health Information System held recently at Georgetown University in Washington DC. PMID- 19562957 TI - Force Health Protection: the mission and political context of the longitudinal health record. AB - Drawing upon an extensive search of publically available literature and discussions at the "National Forum on the Future of the Defense Health Information System," this article documents the evolving mission and political context of the longitudinal health record (LHR) as an instrument for Force Health Protection (FHP). Because of the Gulf War syndrome controversy, the Department of Defense (DoD) launched an ambitious, complex series of programs designed to create a comprehensive, integrated defense health surveillance capability to assure FHP and keep faith with the American people. This "system of systems" includes individual component systems to perform specific functions such as disease surveillance, battlefield assessment, and patient care and consolidates these diverse types of information into centrally accessible archives that serve the interests of occupational health, preventive medicine, medical strategic planning, and longitudinal patient health care. After 25 years of effort and major accomplishments, progress toward a LHR remains uneven and controversy persists. PMID- 19562958 TI - Medical knowledge discovery and management. AB - Although the volume of medical information is growing rapidly, the ability to rapidly convert this data into "actionable insights" and new medical knowledge is lagging far behind. The first step in the knowledge discovery process is data management and integration, which logically can be accomplished through the application of data warehouse technologies. A key insight that arises from efforts in biosurveillance and the global scope of military medicine is that information must be integrated over both time (longitudinal health records) and space (spatial localization of health-related events). Once data are compiled and integrated it is essential to encode the semantics and relationships among data elements through the use of ontologies and semantic web technologies to convert data into knowledge. Medical images form a special class of health-related information. Traditionally knowledge has been extracted from images by human observation and encoded via controlled terminologies. This approach is rapidly being replaced by quantitative analyses that more reliably support knowledge extraction. The goals of knowledge discovery are the improvement of both the timeliness and accuracy of medical decision making and the identification of new procedures and therapies. PMID- 19562959 TI - Software architecture and engineering for patient records: current and future. AB - During the "The National Forum on the Future of the Defense Health Information System," a track focusing on "Systems Architecture and Software Engineering" included eight presenters. These presenters identified three key areas of interest in this field, which include the need for open enterprise architecture and a federated database design, net centrality based on service-oriented architecture, and the need for focus on software usability and reusability. The eight panelists provided recommendations related to the suitability of service oriented architecture and the enabling technologies of grid computing and Web 2.0 for building health services research centers and federated data warehouses to facilitate large-scale collaborative health care and research. Finally, they discussed the need to leverage industry best practices for software engineering to facilitate rapid software development, testing, and deployment. PMID- 19562960 TI - National electronic health record interoperability chronology. AB - The federal initiative for electronic health record (EHR) interoperability began in 2000 and set the stage for the establishment of the 2004 Executive Order for EHR interoperability by 2014. This article discusses the chronology from the 2001 e-Government Consolidated Health Informatics (CHI) initiative through the current congressional mandates for an aligned, interoperable, and agile DoD AHLTA and VA VistA. PMID- 19562961 TI - Interoperability. AB - The main themes of this panel were patient safety and interoperability, the evolution of the traditional longitudinal health record into a patient managed personal health record (PHR) and the personalization of healthcare itself. The interoperability track D of the National Forum posed the following questions: (1) "How do we achieve syntactic interoperability?"; (2) "How do we achieve semantic interoperability?"; (3) "How do we evolve with emerging standards?"; (4) "How do we integrate new medical devices?"; and (5) "How do we achieve National Health Information Network interoperability?" PMID- 19562962 TI - Industry roundtable for interoperability and business process. AB - Healthcare challenges for the military are becoming increasingly complex and the solution to these challenges lies in effective collaboration between industry, government, and academia. Creating and maintaining such collaboration will require political, management, technical, and financial support. This article summarizes a discussion held during the National Forum on the Future of the Defense Health Information System which focused on the best methods for achieving this collaboration and any particular unique roadblocks facing this effort. PMID- 19562963 TI - [Osteoarthritis--a new sight?]. PMID- 19562964 TI - [The occurrence of anticardiolipin antibodies bound in circulating immunological complexes in sera of patients suffering from juvenile lupus erythematosus and juvenile idiopathic arthritis]. AB - BACKGROUND: The presence of antiphospholipid antibodies (APA), especially anticardiolipin antibodies (ACA), antibodies against beta2-glikoprotein I and lupus anticoagulant leads to thrombotic disorders. The pathogenetic role of APA in children is not exactly explained. The frequency of occurrence of APA and antiphospholipid syndrom in children is 2 to 3 times lower than in adults. OBJECTIVES: The aim of the study was to indicate the presence of ACA in circulating immunological complexes (CIC) from sera of patients suffering from juvenile idiopathic arthritis (JIA) and juvenile systemic lupus erythematosus (JSLE). PATIENTS AND METHODS: 31 sera were investigated, 16 from JIA patients and 15 from JSLE patients. The free ACA IgG class, ACA bound in CIC were estimated by the ELISA method (after dissociation of CIC and y-fraction paecipitation). RESULTS: In CIC isolated from sera of JIA and JSLE patients ACA were discovered. ACA, possessing high avidity, in CIC were more frequently discovered than unbound ACA (38.7% vs. 25.8%). CONCLUSIONS: ACA in CIC were more frequent in sera of JIA patients, unbound ACA were more frequent in sera of JSLE patients. PMID- 19562965 TI - [Presence of anticardiolipin antibodies in cryoglobulins isolated from sera of hepatitis C virus infected patients]. AB - INTRODUCTION: Most extrahepatic manifestations of the hepatitis C virus (HCV) infection are connected with influence of virus on the host's immune system. An important part of immunologic disturbances in chronic infections is the presence of circulating immune complexes. Standard serologic methods are not applicable in analysis of complexes composition. OBJECTIVES: Detection of anticardiolipin antibodies (ACA) in cryoglobulins of patients with the HCV infection before and after dissociation of immune complexes. PATIENTS AND METHODS: Sera and croglobulins of 27 patients (19 female, 8 male) with chronic hepatitis C and with cryoglobulinemia were tested. RESULTS: ACA were searched in serum and cryoprecipitate of 27 HCV infected patients with cryoglobulinemia--before and after dissociation of immune complexes. The presence of ACA was not confirmed in sera, but in 59% of cases they were detected in dissociated complexes from cryoprecipitate. The correlation between the presence of anti-HCV antibodies and ACA in cryoprecipitate before dissociation of immune complexes was 0.47 (p < 0.01) and after dissociation--0.61 (p = 0.0007). There was a statistically significant correlation between the presence of purpura (vasculitis) and the presence of ACA before (p < 0.002) and after dissociation of complexes (p < 0.05). CONCLUSIONS: In the tested group of patients there was a high incidence of ACA (non detectable in sera) detectable after dissociation of immune complexes from cryoglobulins. The presence of ACA in cryoprecipitate is significantly connected with clinical signs of cutaneus vasculitis. PMID- 19562966 TI - [Antinucleolar antibodies in diagnostics of antiphospholipid syndrome]. AB - INTRODUCTION: Anti "nucleolar" antibodies are estimated to be present in about 5 10% of anti-nuclear antibodies-positive (ANA), but only in about 15% of cases (sometimes in lower percentages) it is possible to assess precisely the specificity of antibodies responsible for this pattern of ANA. Nucleoli are composed of many proteins, e.g., nucleolin, fibrillarin, Pm-Scl (spliceosome component), RNA-polymerases and different sizes rRNA transcripts. Nucleoli also contain annexins (ANX) II and V. Many of these proteins are target molecules for autoantibodies generated in connective tissue diseases (CTD)--especially in scleroderma and overlap syndromes. OBJECTIVES: The aim of this study was to work out methods for differentiation of antibody specificities in the case of sera ANA positive with the nucleolar pattern in indirect immunofluorescence (IIF), with a particular concern directed to anti-Annexin V antibodies (belonging to anticofactor antibody system--characteristic of antiphospholipid syndrome [APS]). PATIENTS AND METHODS: In the sera of the tested group of 12 (selected from 150 subjects suffering from different kinds of CTD) ANA-positive (Western-blott negative) and showing the nucleolar type of ANA pattern patients, the following autoantibodies were assessed: ANA-IIF, anti-Pm-Scl (overlap syndrome marker), anticardiolipin, antiannexin V, anti-RNA-ase--all using the ELISA method. With the use of the Western-blott method a basic set "marker" autoantibody in CTD was estimated, and the RNA-ase was used for elimination (by enzymatic digestion) of RNA-s and RNA-protein complexes--a possible target for autoantibodies giving the nucleolar pattern. RESULTS: In 5 out of 12 sera (41.7%) of ANA-positive/Western blott-negative CTD patients antibodies against Annexin V, cardiolipin, Pm-Scl were detected. In 2 out of 12 patient sera antibodies to the RNA-ase were found. None of the tested sera showed the presence of autoantibodies against antigen Scl 70 (scleroderma marker), and SS-A (Sjogren syndrome marker), which can also give the atypical nucleolar ANA-pattern. Only in few sera "my ositis"--blotts showed antibodies to antigen Ku (nuclear protein), Jo-1 and PI-7 (anti-tRNA synthetases- polymyositis markers)--myositis and overlap syndrome markers. In 80% of tested sera the co-appearance of anti-ANX-V and anticardiolipin antibodies was observed. The presence of antibodies against ANX-V together with Pm-Scl was confirmed in 60% of sera The RNA-ase, used as a specific rRNA and RNP-protein blocker, resulted in a partial or total disappearance of the "nucleolar" pattern, but independently of the tested serum specificity (anti-ANX-V, anti-Pm-Scl or both together). The observed RNA-ase treatment of the Hep-2 cells effects are equivocal and they may result from either binding or digestion the antigen by the RNA-ase, but also from forming a sterical hindrance for the autoantigen binding to another "nucleolar" antigens. CONCLUSIONS: These preliminary results are interesting, especially as concerns anti-ANX-V autoantigens (associated with APS), but they require further research including bigger groups of CTD patients. PMID- 19562968 TI - [Elderly onset rheumatoid arthritis]. AB - Elderly-onset rheumatoid arthritis (EORA), defined as rheumatoid arthritis (RA) with the onset at the age > or =60 differs sligthy at presentation from younger onset RA (YORA) by a more equal sex distribution, a higher frequency of an acute onset, more frequent involvement of large joints, especially of the shoulder, and higher disease activity. Longitudinal studies have shown greater disease activity, more severe radiographic damage and functional decline in patients with EORA than in those with YORA. These differences were only found in seropositive patients. In seronegative EORA patients a less severe course of the disease and a better outcome have been observed, with the clinical manifestations of polymyalgia rheumatica or remitting seronegative symmetrical synovitis with pitting edema. This article is a review of available data concerning differences between EORA and YORA. PMID- 19562967 TI - [Are rheumatic complaints significantly related to functional ability after 60 year of age?]. AB - INTRODUCTION: Medical care focused on functional abilities gives the elderly with arthritis a possibility of living an independent, active life and participating in social activities, and thus it is seen as the policy of equal chances. OBJECTIVES: The aim of the current paper is the assessment of the Polish adult population mobility level, the changes coming with age, and the statistical significance of relations between rheumatic complaints and a decrease in functional abilities in the elderly. PATIENTS AND METHODS: The results of the all Poland epidemiological research are presented. The survey conducted on a random sample of adult Poles with arthritis took place in 2002. A total of 4017 respondents answered questionnaires prepared by professional investigators. RESULTS: A significant deterioration of functional abilities can be noticed after the age of 40 and it becomes even more severe after 60. A negative assessment of musculoskeletal functions was declared by 70% of the investigated arthritis elderly people. Respondents did not take into account cognitive dissonance when comparing themselves with a peer group. That is why every second patient considered himself mobile enough for his/her age. A low functional assessment relates significantly to the awareness of having arthritis. Higher education increases chances of maintaining functional abilities up to older age. Exercises "for health" are more popular among older men than older women. CONCLUSIONS: Effective preventing the elderly from physical unfitness should be the duty of individuals, medical care and social policy. Popularizing information on the benefits of physical exercises, awareness of the disease, and shifting the time of intervention to the middle age generation can help achieve this aim. PMID- 19562969 TI - [The pharmacokinetic of drugs in elderly patients]. AB - The use of pharmacotherapy in the elderly requires caution because of increasing with age drug sensitivity and risk of dangerous adverse effects. The process of ageing induce alteration in pharmacokinetics of drugs. Modifications affect practically each pharmacokinetic phase. Processes of distribution and, first of all, drug elimination are the most essential for clinical practice. PMID- 19562970 TI - [Polymyalgia rheumatica mimicking neoplastic disease--significant problem in elderly patients]. AB - Polymyalgia rheumatica is a rheumatic disease which mainly affects the elderly, and is seldom diagnosed in patients <50 years of age. The prevalence of polymyalgia rheumatica is approximately 16.8 to 53.7 per 100,000 of the population >50 years of age. Patients may present with spiking fever, malaise, fatigue, weight loss and other features suggesting inflammation, which in each case requires differential diagnosis from malignancies. Neoplastic disease in turn can manifest itself in symptoms resembling those of polymyalgia, which are named "polymyalgia-like syndrome" and are in fact paraneoplastic syndromes presenting as polymyalgia rheumatica. These observations suggest that a careful clinical evaluation and a long term follow-up are necessary for a correct diagnosis of polymyalgia rheumatica. PMID- 19562971 TI - [Social and economic aspects of osteoarthritis]. AB - Osteoarthritis (OA) is the most common form of arthritis. The incidence of the disease increases with age. Demographic forecasts predict aging of the population and an increase in the obesity rate in the society, the factors that contribute to the development of osteoarthritis. Thus, the nearest future will most likely bring an epidemic of the disease. While estimations of direct and indirect costs of OA have been developed in many countries, no such analyses have been made available for Poland. The total cost associated with disability pensions, hiring professional care and general loss of work productivity exceeds the direct cost of diagnostics and treatment. On a country scale, OA and its consequences require significant expenditure on the part of the health care system, social security system and the national budget. Optimization of this cost will require completion of epidemiological and pharmacoeconomic analyses and transformations in the health care and social security systems. The job market will also need to be adapted to the inevitable changes by means of introducing necessary legal adjustments to facilitate continued employment of persons >50 years of age with musculoskeletal disorders. PMID- 19562972 TI - [Soft tissue rheumatism in erderly]. AB - Disorders of soft, peri-articular tissues are a common cause of musculoskeletal pain in elderly patients. Nevertheless, most physicians underestimate the role of soft tissue rheumatism in the pathomechanism of the pain. The impairments of soft tissue can not be diagnosed by X-rays examinations, whereas degenerative lesions of joints are easy diagnosed using this method even despite of their uncertain role in producing the symptoms. The incidence of pain syndromes originated from soft tissues differ regarding to the age of patients. In young subjects the incidence of all of them is generally low. Syndromes provoked by overloading during work: repetitive strain syndrome, canal tunnel syndrome, tennis elbow, golfers elbow, shoulder tendon coin disorders and myofascial pain syndrome are common in middle-aged patients. The morbidity of fibromialgia syndrome is also lower in old people probably as the result of diminished numbers and degenerative changes in nociceptive fibers. The syndromes prevailing in elderly patients include trochanteric syndrome and the pain syndromes provoked by muscle spasm depended on posture abnormalities. In the soft tissue pain syndrome prevention adapted to old age kinesitherapy and avoiding muscle overloading are recommended. Soft tissue pain syndromes are usually treated with non steroidal anti inflammatory drugs. In local pain syndromes better results can be obtained by local treatment. Local injections of glikocorticosteroids are usually very effective and safe. PMID- 19562973 TI - [Senile osteoporosis]. AB - Osteoporosis is a skeletal disease characterized by the increased risk of fractures, decreased bone mass and bone quality. The number of patients suffering from osteoporosis still increase due to the extension of the lifespan in the developed countries. The low-energy hip fractures are the most severe complications of the disease, lethal in some cases. The number of hip-fractures in Poland exceeds 30,000 per year. Senile osteoporosis makes 20% of primary osteoporosis. The calcium and vitamin D3 intake from diet or their supplementation are crucial in the ethiopathogenesis of this disease. The effective treatment should include non pharmacological methods, fall prevention, and individually chosen pharmacological treatment. PMID- 19562975 TI - [Clinically asymptomatic antiphospholipid antibodies--diagnostic and therapeutic problems]. AB - The presence of asymptomatic antiphospholipid antibodies (aAPA) creates many difficult diagnostic and clinical problems - they might be a cause either of incorrect hemorrhagic diathesis recognition or an unnecessary anticoagulant therapy. Patients with aAPA should be counseled individually regarding potential thrombotic and/or obstetric complications. An increased risk of these complications is particularly high when lupus anticoagulant (LAC) and IgG anti beta2 glycoprotein antibodies coexist. The approaches to the diagnosis and management of treatment of patients with aAPA are also discussed. PMID- 19562974 TI - [Organ alterations due to aging]. AB - Aging defined as progressive organ dysfunction which makes keeping homeostasis more difficult starts at the age of 30-40. However, due to difficulties with the distinction between aging and disease processes, changes previously believed to be caused by aging are often recognized as the effect of pathologies when new data is presented. According to current knowledge, cardio-vascular aging includes decreased elasticity of main arteries, decreased ability of left ventricule to relaxate, diminished function of sino-atrial node and decreased effect of beta adrenergic stimulation. Aging in the respiratory system is attributed to increased size of alveoli and alveolar ducts (which easier collapse), a decrease in the gase exchange area, a decrease in respiratory volumes (both static and dynamic) and a severe decrease in maximal oxygen consumption. In aging kidneys both renal blood flow and glomerular filtration are decreased. As the result of tubular alterations the kidney ability to conserve and dilute urine, and its capability to regulate the pH and serum sodium level diminish. Less dramatic changes are seen in the gastrointestinal tract. According to available data, high prevalence of gastric atrophy and hypochlorhydria are a consequence of Helicobacter pylori infection. Also, constipation is attributed much more to sedative life style and diet than to aging itself. In summary, none of the presented alterations is severe enough to cause the disease, but all of them increase the risk of pathology and thus pave the way for the disease even in healthy elderly subjects. PMID- 19562976 TI - [Chorea as a rare symptom of systemic lupus erythematosus in the elderly]. AB - The opinion that systemic lupus erythematosus (SLE) is a young people disease translates into its rare diagnosing in subjects in their 6th decade of life and older. The variability of neuropsychiatric disorders that may be the first symptoms of the disease suggest the need for their in-depth diagnostics and treatment. Chorea is one of the rarest neuropsychiatric symptoms of SLE. It may be present in an early stage of the disease and it is one of the most common motor disorders in SLE. The presented case of diagnosing SLE in a 79-year-old male shows the importance of individual and thorough assessment of the clinical picture of every patient regardless of his/her age and gender. In the discussed case the clinical course of the disease denies common opinions about the picture of SLE in the elderly. PMID- 19562977 TI - [Symptoms and signs of polymositis, systematic lupus erythematosus and antiphospholipid syndrome follow consecutively]. AB - Polymyositis (PM) and dermatomyositis (DM) are very rare connective tissue disorders which only in exceptional circumstances affect white men. The present paper describes the case of an obese 55-years-old man in whom no muscular skeletal system symptoms were found during the period of 2 years before the onset of arthritis, and who was treated because of cardiac involvement (pericarditis, PAF), pleuritis, malaise and fever. Only the occurrence of non erosive arthritis decided on the connective tissue disorder as a cause of his complaints. Previously the diagnostics was based on the exclusion of malignant diseases and bacterial and viral infections. The determination of antinuclear antibodies in high titer without any specific, typical of individual disease entities antinuclear antibodies allowed only the diagnose of undifferentiated connective tissue disease. Glucocorticosteroid treatment was initiated, however six months later despite treatment with metyloprednisolone the symptoms and signs associated with PM accompanied by high level of creatinophosphokinase and elevated transaminase were found. The muscle biopsy revealed myositis, but no specific antibodies, especially anti-Jo-1, were detected. It is very interesting that cyclophosphamide + glucocorticoids pulse therapy failed to prevent the development of antiphospholipide syndrome and interstitial pulmonary fibrosis in the PM patient. Overlapping of immunological tissue disorders is a well-known phenomenon, but in the case of fulminating and distinct symptoms and signs the lack of detection of specific antibodies is quite uncommon. PMID- 19562978 TI - Early defibrillation: a key for successful outcome of in-hospital cardiac arrest. AB - BACKGROUND: The objectives of this study were to determine 1) the rate of delayed defibrillation and 2) the importance of early defibrillation in patients with cardiac arrest who need defibrillation in a large tertiary care hospital. MATERIAL AND METHOD: We analyzed data from Siriraj cardiopulmonary resuscitation (CPR) registry from January 2005 to December 2007. The registry recorded setting and cause of cardiac arrest, timing of cardiac arrest and time initiation of each step of treatment such as basic life support (BLS), advanced life support (ALS), defibrillation, medication, time of defibrillation. Outcome was recorded as return of spontaneous circulation (ROSC) which lasted at least 20 minutes and discharge from hospital. RESULTS: A total of 2160 in-hospital cardiac arrest records were sent to CPR center and were evaluated. 612 patients (28.3%) needed defibrillation. Average age was 57.1 +/- 21.2 years. Among patients who needed defibrillation, 250 patients (40.8) had early defibrillation. Median time to defibrillation after the detection of cardiac arrest was 8 (3, 15) minutes. Factors associated with delayed defibrillation were the patients being in non intensive care unit (non-ICU) wards, being in wards without standby defibrillator, and female gender. 283 patients (46.2%) had ROSC after CPR and 50 patients (8.2%) survived to discharge from hospital. Time to defibrillation was the most important predictor for ROSC and survival to discharge. CONCLUSION: Among in-hospital patients with cardiac arrest and who needed defibrillation, early defibrillation is the major key to a successful outcome. PMID- 19562979 TI - The effect of a continuity of care clinic curriculum on cardiovascular risk management skills of medical school graduates. AB - BACKGROUND: The continuity of care clinic (CCC) curriculum has been added to final-year medical students, class of 2008. The goals were to improve cardiovascular risk management skills for medical students and to develop competent doctors to serve the public. OBJECTIVE: To study the effectiveness of the curriculum by directly comparing postgraduate patient care performance between CCC participants (class of 2008) and non-CCC participants (class of 2006 and 2007). MATERIAL AND METHOD: We collected information about both groups of graduates, when they started their doctor careers. With hospitals'permission, medical charts audits were performed and scored with a 12-task checklist of cardiovascular risk management. The scores from both groups were compared with statistical analyses. RESULTS: Among 266 charts from 17 hospitals, there were 123 charts from 38 CCC participants and 143 charts from 52 non-CCC participants. On 9 of 12 tasks of the checklist, proportionately more CCC participants carried out the tasks than non-CCC participants. Statistical significance was shown on 5 tasks. These were ability to properly adjust antihypertensive medication (13.4% more; p = 0.002); requesting for urine protein screening (12.1% more; p = 0.006); recommending life-style modification (24.9% more; p < 0.001); requesting for serum lipid profile (25.5% more; p < 0.001); prescribing aspirin as primary prevention for cardiovascular disease (13.1% more; p = 0.007). There was no statistically significant difference for the other 7 tasks. CONCLUSION: Cardiovascular risk management performance of CCC participants was better than non-CCC participants in the same period after graduations. The curriculum helped improve the cardiovascular risks management skill of postgraduates. In the public interest, this study recommends further implementation of such a program in the future. PMID- 19562980 TI - Safety and tolerability of galantamine in possible Alzheimer's disease with or without cerebrovascular disease and vascular dementia in Thai patients. AB - The purpose of this study was to explore factors that influence the clinical safety and tolerability associated with galantamine administration in Thai Alzheimer's disease patients with or without cerebrovascular disease and vascular dementia. This was an analysis of previous study. Tolerability and safety profile were analyzed according to sex, age, body weight, Thai mental state examination (TMSE) score, Alzheimer's disease assessment scale-cognitive subscale (ADAS-cog) score, and Alzheimer's disease cooperative study/activities of daily living (ADCS/ADL) score. The most common adverse events were nausea, dizziness, and weight loss which more often occurred during the dose-escalation phase. Mean body weight lost at week 24 was 0.9 kg. Sex, age, body weight, and ADAS-cog score did not influence the incidence of any adverse events. Dizziness was more likely to occur in patients with low TMSE and high ADCS/ADL score (p = 0.02 and p = 0.050, respectively). Patients with TMSE score equal or higher than 23 more often experienced muscle cramps and fatigue than who had TMSE lower 23 (p < 0.05). However, flexible dose escalation of galantamine with a 4-week schedule was safe and well tolerated in Thai AD patients. PMID- 19562981 TI - Outcome of medical thoracoscopy. AB - BACKGROUND: Medical thoracoscopy is a common procedure for pulmonologists working in Europe but is still unpopular in South East Asia with few medical centers in Thailand performing this procedure. We report our outcome of medical thoracoscopy based upon 10 years experience. MATERIAL AND METHOD: Medical thoracoscopy was first performed in our unit in 1998. The early indication was undiagnosed pleural effusion. Other indications which were empyema, pneumothorax and talc pleudrage were carried on for the last 3 years. The patients' demographic data, indication for medical thoracoscopy, procedures, complication and outcome were recorded and analyzed separately. RESULTS: During 1998 to 2007, there were 142 procedures of medical pleuroscopy performed. There were 86 procedures for the indication of undiagnosed pleural effusion. The diagnostic yield was 95.2%. The malignancy was recovered by thoracoscopy in 45.35% of procedure. For indication of talc pleurodesis, there were 22 patients with 3 who had early failure of pleurodesis because of trapped lung. After mean follow up of 124 days, 17 patients did not have recurrence of pleural effusion. 15 patients who had loculated pleural effusion were done medical thoracoscopy. Operations were successful in only 6 patients. For indication of pneumothorax 5 out of 6 procedures were successful after mean follow up of 167 days. In 12 empyema patients, mean hospital admission was 9.1 days after thoracoscopy. There was no serious complication from the procedure. DISCUSSION: There are many indications for medical thoracoscopy and the experience of the performer is the important factor determining success of the procedure. In undiagnosed pleural effusion, our result was comparable to other studies in the past in which the rate of malignancy was around 40-60%. The result of talc pleudrage was also comparable with the need to improve the diagnosis of trapped lung to prevent the unnecessary medical thoracoscopy. The result was excellent in patients who came for pleurodesis indicated in pneumothorax also in empyema but number of patient was still low. The problem was in loculated pleural effusion from malignancy which showed high failure rate. The early pleurodesis in malignant pleural effusion before it became loculated should be considered. CONCLUSION: Outcome of medical thoracoscopy varies from various indications. The success rate was high and decreasing in undiagnosed pleural effusion, pneumothorax, empyema and talc pleurodesis in malignant effusion. Great skill was needed to perform medical thoracoscopy in loculated malignant pleural effusion PMID- 19562982 TI - Is cytology necessary in diagnosis of mediastinal mass? AB - BACKGROUND: Mediastinal mass is an uncommon abnormality found in clinical practices of respiratory physicians. The diagnosis of a mediastinal mass arises from a clinical suspicion, and more commonly, by a check-up chest x-ray. Definite diagnosis is necessary because the managements are different in various etiologies of the masses. OBJECTIVE: Adequacy of cellular sample recovered from small needle aspiration is the point of question among pathologist and physician in the diagnosis of mediastinal mass. Many centers recommend fine needle aspiration biopsy for cytology (FNAB) as an adequate procedure in this situation. This study is aimed to find the value of cytology and histological examination in the diagnosis of a mediastinal mass. MATERIALS AND METHOD: The study was done by prospective collected data of patients who were consulted for needle aspiration biopsies of their mediastinal mass since 1999 to 2006 at the Respiratory Diagnostic Unit, Division of Respiratory disease and Tuberculosis, Faculty of Medicine Siriraj Hospital. The protocol in evaluating mediastinal mass was to obtain both cytologic slides and tissue for histology from lesions in the mediastinum by ultrasonic guidance when they were possible. RESULTS: During 1999 to 2006, there were 35 patients who had mediastinal masses referred for needle aspiration procedures. Their mean age and standard deviation were 42.37 +/- 16.97 year-old. Among these patients, 22 were men and 13 were women. The mean age (+/- standard deviation) of male patients was 40.47 +/- 17.17 years and 45.5 +/- 16.79 year-old for the female. The histology could make diagnosis in a significantly higher number of patients as compared to cytology (88.57% compared to 40%). There was statistically different higher diagnostic rate of UG-CNB than UG-FNA in non carcinomatous group. The complications were mild. CONCLUSION: Since most lung mass are carcinomatous in origin, depending on various factors (age, sex, smoking habit and size), the recommendation of FNA as the first line investigation are warranted because of high yield to risk for complication ratio, but in mediastinal mass in which many etiologic cells of origin cannot be diagnosed accurately by retrieved cytological cell, the CNB for small histology section is recommended as an initial investigation method without the loss of time required for cytological aspiration. PMID- 19562983 TI - Asthma, bronchial hyper-responsiveness and Chlamydophila (Chlamydia) pneumonia infection in adult Thai population. AB - BACKGROUND: The associations between Chlamydophila (Chlamydia) pneumonia infection and chronic asthma or bronchial hyper-responsiveness (BHR) have been inconclusive. OBJECTIVE: We aimed to determine the association between C. pneumonia infection and asthma as well as BHR in the adult Thai population. MATERIAL AND METHOD: This nested case-control study retrieved the data from a nation-wide Respiratory Health Survey (2001-02) in the adult population (age 20 44 year) in Thailand. Each subject underwent questionnaire interview, spirometry, bronchoprovocative test, skin prick test for common aeroallergens and venous blood collection. Subjects with BHR (n = 79) including those with asthma (n = 52), were randomly selected as cases. Subjects without BHR or asthma were also randomly selected as the control (n = 137). We used the stored serums for the C. pneumonia serologic assay including IgA, IgG and IgM by microimmunofluorescence (MIF) technique. RESULTS: There is no significant relationship between chronic Chlamydia infection (IgG > or = 1:512 and IgA > or = 1:40) and BHR or asthma. Higher IgM was found in subjects with BHR when compared with the control group (p = 0.04). The IgM titer > or = 1:10 was associated with BHR with borderline significance (odds ratio 1.98; 95% CI 0.98-4.00; p = 0.05). Logistic regression analysis revealed no evidence of confounding effects for age, sex and atopy. However mite allergy seems to be an effect modifier of the relationship between the recent Chlamydia infection and BHR. CONCLUSION: The present study does not support the hypothesis about the association between persistent C. pneumonia infection and chronic asthma. However the recent infection may be related with bronchial hyper-responsiveness particularly in those without allergy to house dust mite. PMID- 19562984 TI - Changing of the etiology of acute pancreatitis after using a systematic search. AB - BACKGROUND: Alcohol and gallstone are the 2 most common etiologies of acute pancreatitis (AP). In Thailand, alcohol has been believed to be the leading etiology of AP. However, a thorough and systematic search may discover real etiology of AP. MATERIAL AND METHOD: During 2006 to 2007, seventy-eight patients with AP were prospectively searched for the etiology by: 1. Performing liver chemistry tests and transabdominal ultrasonography (US) for gallstone in every case; 2. Measuring serum triglyceride and calcium in every case; 3. Investigating definite drugs use or other identified etiology; 4. Asking about the amount of alcohol ingestion (amount > 80 g/day for > 5 years was required for alcoholic AP; 5. Performing CT scan (if age > 40 years) and EUS if no etiology was identified. Results were compared with the retrospective data from medical records of 66 AP patients during 2003-2005. RESULTS: Of the 78 patients, the etiologies were alcohol in 32 (41%), gallstones in 29 (37%), miscellaneous in 13 (17%) and idiopathic AP in 4 patients (5%). When compared with the retrospective data of the 66 patients over the past 3 years, the etiologies were alcohol 53%, gallstone 22%, miscellaneous 11% and idiopathic 14%. Among the 45 patients of the study period (58%) who consumed alcohol more than the defined threshold for alcoholic AP, 13 (29%) were found to have other explainable causes of AP, i.e gallstones in 10, hypertriglyceridemia in 2 and AIDS cholangiopathy in 1 patient. CONCLUSION: Alcohol was probably over-diagnosed as a leading etiology of AP in the past. A systematic search of the etiologies lowered the frequency of alcoholic and idiopathic AP but discovered more patients with gallstone pancreatitis. One fourth of AP patients who were heavy drinkers had other explainable etiologies of AP. PMID- 19562985 TI - Natural course of abdominal pain in chronic pancreatitis with intermittent (type A) pain after conservative treatment. AB - BACKGROUND: Abdominal pain in chronic pancreatitis (CP) is difficult to treat and appropriate choice of treatment is controversial. It has been suggested that patients with CP particularly from alcohol (ACP) with intermittent attack of abdominal pain (type A pain) should be managed conservatively because pain relief will be achieved in most cases. However, data of the efficacy of this strategy is scanty and conflicting and whether this strategy is effective or feasible in idiopathic CP (ICP) is unclear. MATERIAL AND METHOD: Data of all patients with CP with type A pain, who were followed-up and managed conservatively during 2004 2008 were analyzed. Pain relief was defined by the absence of abdominal pain for more than 1 year RESULTS: Twenty-two patients were followed-up with a median duration of 31 months (range 5-96 months). The etiology of CP was alcoholic (ACP) in 12 (56%), early-onset idiopathic (E-ICP) in 5 (22%) and late-onset idiopathic (L-ICP) in 5 (22%). Alcohol abstinence was successful in every ACP patient. Overall, 18 patients (82%) had pain relief with a median duration of 39 months (range 16-167 months) from the onset of pain or 14 months (range 11-57 months) from the time of diagnosis of CP. Pain relief was achieved at a higher level mainly in ACP (100%) and L-ICP (80%) but was only 40% in E-ICP Median duration from onset until pain relief were 28 months (range 16-167 months) for ACP, 36 months (range 16-39 months) for L-ICP and 120 months (range 42-120 months) for E ICP. The difference was statistically significant between L-ICP and E-ICP (p = 0.036), but not between ACP and E-ICP (p = 0.13) and between ACP and L-ICP (p = 0.80). Median duration from the time of diagnosis of CP until pain relief was only 14 months forACP 13 months for L-ICP but was 52 months for E-ICP. None of the patients required narcotics, endoscopic therapy or surgery. CONCLUSION: Conservative management was feasible and effective in most patients with CP and type A pain, particularly ACP after alcohol abstinence, and L-ICP Conservative treatment was not effective in E-ICP. PMID- 19562987 TI - Non-diabetic glomerular disease in type II DM: 10 years experience. AB - BACKGROUND: The nature of renal damage in patients with type II diabetes remains unclear. OBJECTIVE: To analyze the renal histopathology in type II diabetes who underwent renal biopsy at Siriraj Hospital, renal unit over 10 year period. MATERIAL AND METHOD: The clinical and biochemical data in 54 patients with Type II DM, atypical cases of DN, were subjected to renal biopsy and analyzed retrospectively. RESULTS: Ten out of fifty-four type II diabetic patients (18.5%) were diagnosed non-diabetic nephropathy (NDN); there were 4 patients with membranous GN, 3 patients with crescentic GN1 patient of MPGN type I, 1 patient with renal change from hypertension and 1 patient with IgMN. The most important factor that had statistically significant was nephritis urine sediment (NDN: DN 40% vs. 4.5%), However 60% of NDN had no nephritic urine sediment. CONCLUSION: There was no strong predictor to differentiate DN from NDN by clinical or biochemical data. The only significant finding in NDN was nephritic urine sediment. PMID- 19562986 TI - Clinical utility of lens culinaris agglutinin-reactive alpha-fetoprotein in the diagnosis of hepatocellular carcinoma: evaluation in a Thai referral population. AB - BACKGROUND AND OBJECTIVE: There is no established clinical role for the lens culinaris agglutinin-reactive fraction of alpha-fetoprotein (AFP-L3%) in the management of the Thai hepatocellular carcinoma (HCC) patient population. The aim of this prospective study was to evaluate clinical utility and performance characteristics of AFP-L3% for the diagnosis of HCC in Thai referral patients. MATERIAL AND METHOD: Sixty-one histologically proven HCC patients and 35 patients with other liver cancers were included for analysis. RESULTS: The HCC population was comprised of 50 males and 11 females, with a mean age of 48.8 years. According to the Okuda system, three were classed as stage I, thirty-five belonged to stages II, and six were classified in stage III. An AFP-L3% a cut-off value of > 15% yielded a sensitivity of 82% (95% confidence interval [CI], 74 88%), specificity of 71% (95% CI, 58-82%), positive predictive value of 83% (95% CI, 75-90), and negative predictive value of 69% (95% CI, 56-80) for the diagnosis of HCC. In HCC patients with AFP of < 200 ng/ml, an AFP-L3% at a cut off value of > 15% not only maintained high sensitivity of 83% and good specificity of 71% but also increased negative predictive value to 86% for the diagnosis of HCC. CONCLUSION: AFP-L3% provides high sensitivity but with lower sensitivity in the diagnosis of HCC than total AFP in individuals with symptomatic liver mass. However, considering its high negative predictive value in patients with AFP < 200 ng/ml, AFP-L3% might be useful as an adjunctive marker, in combination with AFP, to exclude the presence of HCC. PMID- 19562988 TI - The clinical outcome of acute kidney injury in critically ill Thai patients stratified with RIFLE classification. AB - OBJECTIVE: The Acute Dialysis Quality Initiative (ADQI) Group published a consensus definition (the RIFLE criteria) for acute renal failure. We sought to assess the ability of the RIFLE criteria to predict mortality in critically ill Thai patients with acute kidney injury (AKI). MATERIAL AND METHOD: We performed a retrospective cohort study, in Siriraj Hospital (a large single tertiary care academic center in Thailand) on 121 patients admitted during November 2005 November 2006. We classified patients according to the maximum RIFLE class (class R, class I or class F) reached during their hospital stay. Demographic data, hospital mortality, hospital length of stay, and need of renal replacement therapy was collected. RESULTS: Patients with maximum RIFLE class R, class I and class F had hospital mortality rates of 35.7%, 35.7% and 65.9%, respectively, compared with 20% for patients without acute kidney injury. Overall hospital mortality of the patients in AKI group (Risk, Injury, Failure group) was increased when compared with no AKI group (Odds ratio = 4.2; 95% Confidence Interval, 1.6-10.6; p =0.003). Mortality was not significantly different among those with the "Risk" and "Injury" class of RIFLE AKI compared with those without AKI, but mortality increased significantly with the "Failure" class (Odds ratio = 7.7; 95% Confidence Interval, 2.7-21.8; p < 0.001). There was the highest rate of renal replacement therapy in the failure group (52.3%) compared with no AKI group (5.7%), and injury group (7.1%) (p < 0.001). CONCLUSION: Acute kidney injury 'risk, injury, failure', as defined by the newly developed RIFLE classification, is associated with increased hospital mortality and renal replacement therapy in critically ill Thai patients. PMID- 19562989 TI - Epidemiology of sepsis in Siriraj Hospital 2007. AB - BACKGROUND: Sepsis remains a major health burden, and there is limited epidemiological report of sepsis in Thailand. OBJECTIVE: To determine the epidemiology, treatments, clinical courses and outcomes of sepsis patients. MATERIAL AND METHOD: All sepsis patients in medical wards at Siriraj Hospital were recruited from February 1 to July 31, 2007. The information from patients' medical records were retrieved and analyzed. RESULTS: From 3,451 patients, 201 (5.8%) were diagnosed as sepsis, and 38.8% of these developed septic shock. Among sepsis patients, 62.2% were community acquired, 40.8% had bacteremia, and gram negative bacteria were the common pathogen (51.7%). Appropriate antibiotics were given within 6 hours in 39%. Goal-directed therapy was achieved in only 11.5%. The mortality among sepsis and septic shock patients was 34.3% and 52.6%, respectively (p = 0.008). Risk factors for hospital mortality included higher maximum SOFA score, hospital-acquired infection, central nervous system dysfunction and receiving antibiotics after 6 hours of onset of sepsis. CONCLUSION: Sepsis is still common and has contributed to high mortality. Goal directed therapy and appropriate antibiotics given within 6 hours might improve the outcome. PMID- 19562990 TI - Serum procalcitonin in diagnosis of bacteremia. AB - OBJECTIVE: To (a) determine the diagnostic value of procalcitonin (PCT) in differentiating sepsis with or without bacteremia, (b) evaluate the correlation of PCT levels to severity of sepsis, (c) establish the prognostic value in predicting the outcome of sepsis and (d) evaluate the correlation among different assays. MATERIAL AND METHOD: A prospective study was carried out from August through November 2007. Blood for PCT levels and culture were drawn simultaneously. RESULTS: Fifty-six patients with clinical suspicious of sepsis were enrolled in the study; bacteremia (n = 30) and non-bacteremia (n = 26). There were good correlations between the PCT levels measured by three assays (p < 0.001). At the threshold of 0.5 ng/mL, PCT had > 90% sensitivity for diagnosis of bacteremia. Of the bacteremic group, median PCT levels measured by Kryptor and VIDAS assays were 12.4 and 16.6 ng/mL respectively. In the non-bacteremic group, median PCT levels measured by Kryptor and VIDAS were 4.2 and 4.9 ng/mL respectively. PCT levels were significantly higher in the bacteremic group (p = 0.04). The optimum thresholds to discriminate between these two groups were found to be 5, 6.5 and 2 ng/mL for Kryptor, VIDAS and PCT-Q, respectively. In addition, correlations of PCT and increasing values of the APACHE II score were observed. PCT levels in the severe sepsis and MOD group were also found to be significantly higher CONCLUSION: PCT was highly sensitive in detecting bacteremia, although not very accurate in differentiating bacteremic from non-bacteremic SIRS in adult patients. PMID- 19562991 TI - Stress hyperglycemia in patients with sepsis. AB - BACKGROUND: Patients with acute stress from stroke or myocardial infarction may develop hyperglycemia, even in the absence of diabetes mellitus. Stress hyperglycemia increases morbidity and mortality in these patients. There has been no study to evaluate stress hyperglycemia in patients with sepsis. MATERIAL AND METHOD: A prospective cohort study in patients with sepsis admitted in the Department of Medicine of Siriraj Hospital during 2006-2007 was done. Data were collected on admission blood glucose, HbA1c and other factors which may predict the outcomes of sepsis. RESULTS: Data were collected from 70 patients with sepsis. The prevalence of stress hyperglycemia was 42.3% in this study. We found no differences in clinical findings, laboratories, interventions and outcomes between groups of stress and non-stress hyperglycemia. Multivariate analysis showed that only APACHE II score and use of a mechanical ventilator were associated with mortality. CONCLUSION: The prevalence of stress hyperglycemia in patients with sepsis was high. We cannot conclude that stress hyperglycemia did not affect the mortality and morbidity outcome mainly because of the small number of subjects which may be not enough to detect statistical significance. PMID- 19562992 TI - Retrospective study of patients with herpes simplex encephalitis and positive CSF PCR. AB - OBJECTIVE: To determine the clinical presentations, laboratory and imaging findings of patients with HSE and positive CSF PCR then compare these with information from other viral encephalitis patients in order to find clinical clues that might help clinicians in early diagnosis and treatment of HSE while awaiting for the CSF PCR result. MATERIAL AND METHOD: A patient group with both HSE and positive CSF PCR and a control group of patients with other viral encephalitis were identified from Siriraj Hospital database within the period of 1997-2006. Medical notes of these patients were reviewed and relevant information, including clinical, laboratory and imaging study, were extracted. Then, descriptive statistics, unpaired t-test and Fisher's exact test were performed with the purpose to determine any clinical or laboratory clues that are significantly different between these two groups, which might help clinicians in making an early diagnosis of HSE. RESULTS: Seven HSE cases and 22 cases in control group were included. Fever, headache, alteration of consciousness, behavioral change, neck stiffness are the most commonly found clinical presentations in HSE patients with CSF PCR positive. CSF examination show CSF leukocytosis with lymphocyte predominate, decreased CSF:blood glucose ratio and elevated CSF protein in all cases. However, no single clinical or laboratory finding helps in differentiating HSE with positive CSF PCR from other viral encephalitides, except radiological abnormalities of temporal lobe, which yields a positive predictive value of 0.5 and a negative predictive value of 0.93. CONCLUSION: Our study showed that HSE shares common clinical and laboratory findings with other viral encephalitides, except for temporal lobe involvement. Early diagnosis is important and empirical acyclovir should be commenced early in patients with viral encephalitis, especially for those with radiological abnormalities on the temporal lobe. PMID- 19562993 TI - The study of clinical manifestation of osteoarticular tuberculosis in Siriraj Hospital, Thailand. AB - OBJECTIVE: To study of clinical manifestations of osteoarticular tuberculosis. MATERIAL AND METHOD: This study is an observational retrospective study design from the electronic medical record database of Siriraj Hospital in 2005-2006. RESULTS: Ninety- nine patients fit in to the inclusion criteria. All were non HIV infected. There were 44 males (44.4%) and 55 females (55.6%) with a male/female ratio of 8:10. The mean and median age was 50.9 +/- 19.8 and 50 years, respectively, with a range from 1-85 years. The most frequent chief complaint was pain (84.8%), followed by functional disability/neurological deficit (47.5%), constitutional symptoms (38.4%), fever (24.2%), localized swelling (21.2%), fistulas and drainage (11.1%) and discovery as a co-morbid disease (1%). The most frequent clinical manifestation was tuberculous spondylodiscitis (66.7%) followed by tuberculous septic arthritis (28.3%), tuberculous osteomyelitis (10.1%), tuberculous tenosynovitis (4.0%), tuberculous bursitis (2.0%) and tuberculous pyomyositis (2.0%). Concurrent pulmonary TB was 30.3%. The mean and median time to diagnosis was 20.4 +/- 16.9 and 13 weeks, respectively, with a range from 4 days to 104 weeks. The shortest duration of time to diagnosis was 0-4 weeks (19.2%) followed by 12-16 weeks (14.1%) and 20-24 weeks (13.1%). The longest duration of time to diagnosis was 104 weeks (1%). The diagnosis from histopathology was 46.5%, positive AFB 40.4%, positive PCR 33.3% and positive culture 19.2%. The radiological abnormalities were osteolytic lesion (79.8%), narrowing intervertebral disc space/joint space (54.5%), cold abscess (49.5%) and vertebral collapse (43.4%). Surgery with medical treatment was performed on 72.7% while the percentage of those receiving medical treatment only was 27.3%. The results of treatment were improvement with residual deformities/dysfunction occurring in 97%. Death occurred in 3%. CONCLUSION: Osteoarticular tuberculosis is common in all age groups. Tuberculous spondylodiscitis was the most frequent clinical manifestation in Siriraj Hospital, Thailand. Although the patients were provided with adequate medical and surgical therapy, osteoarticular tuberculosis was still associated with mortality and morbidity. PMID- 19562994 TI - Cancer anemia survey in Division of Medical Oncology at Siriraj Hospital (CAS). AB - BACKGROUND: Causes of anemia in cancer patients are multifaceted and include such factors as nutritional deficiency anemia, anemia of malignancy and treatment related anemia. Chemotherapy, especially a platinum-based regimen, is well recognized to cause anemia. Anemia results in decrease of functional capacity, lower performance status, poor compliance and adverse prognosis. Optimal management of anemia in cancer patients is an essential component of cancer treatment. Previously, there was no information about this condition available in Thailand. OBJECTIVE: 1) To evaluate frequency, characteristics of anemia and treatments of anemia in our cancer patients who received chemotherapy treatment. 2) To identify the factors that influence hemoglobin (Hb) level in cancer patients especially chemotherapy regimens. MATERIAL AND METHOD: Patients with diagnosis of solid malignancy who were scheduled to receive chemotherapy for least 4 cycles between June 2006 and December 2007 were included. All enrolled patients' data which included demographics, types and stages of cancer, chemotherapy regimen, Hb at baseline, Hb level before each cycle of chemotherapy and treatment of anemia, were recorded. STATISTICAL ANALYSIS: Patients' data were presented in terms of percent, mean or median. Logistic-regression analysis was performed to identify risk factors of anemia in cancer patients. RESULTS: Three hundred and four patients were enrolled, 233 patients were female and 71 were male. The age of patients varied from 15 to 86 years old. Median age was 52 years old. Hb level at enrollment ranged from 7.7 to 16.1 g/dl, mean baseline Hb was 12.5 g/dl. Incidence of anemia at baseline was 34.5 percent whereas the incidence increased to 61.1 percent after receiving chemotherapy. The incidence of anemia in all patients was not significantly different from that of a subgroup of patients with normal baseline Hb. The patients who received platinum and anthracycline-based chemotherapy developed more anemia than those who received other chemotherapy regimens, with odd ratios of 9.4 (95% CI; 3.1-28.9, p < 0.001) and 3.5 (95% CI; 1.4-8.5, p = 0.005), respectively. Most anemic patients were asymptomatic; twenty-one out of 214 anemic patients (9.8%) received specific treatment for anemia. CONCLUSION: Chemotherapy-induced anemia is a common problem found in cancer patients, especially in those receiving platinum-based chemotherapy Most of the anemic patients had asymptomatic grade I and II anemia. Blood transfusion was the treatment of choice for severe, symptomatic anemia in our hospital. PMID- 19562995 TI - Arterial occlusion in nephrotic syndrome: report of four cases in Siriraj Hospital. AB - Venous thrombosis is commonly found in nephrotic syndrome, but arterial occlusion is never report in Thailand. Four cases with cerebral and femoral arteries occlusion were demonstrated. The early diagnosis and appropriate intervention can improve outcomes, reduce mortality and morbidity significantly. PMID- 19562996 TI - [The new thought for research of spleen function]. PMID- 19562997 TI - [Splenic autotransplantation combined with lower esophagus transaction in the treatment of hepatic cirrhosis induced portal hypertension]. AB - OBJECTIVE: To study the therapeutic effect of splenic autotransplantation combined with lower esophagus transaction anastomosis in the treatment of liver cirrhosis induced portal hypertension. METHODS: Thirty-six patients admitted from January 2003 to December 2006 were randomly divided into splenic autotransplantation group undergoing splenic autotransplantation after splenectomy combined with lower esophagus transaction anastomosis, and splenectomy group only undergoing splenectomy combined with lower esophagus transaction anastomosis. The general conduction, splenic scanning, liver function, and the level of serum Tuftsin and IgM of each patient were observed before and after operation. RESULTS: The levels of Tuftsin and IgM in splenic autotransplantation group were significant higher than that of splenectomy group 2 months after the operation, and the liver function showed no significant difference between these two groups. Splenic tissue was detected in the retroperitoneal space by 99mTc-DRBC 2 months after operation. CONCLUSIONS: Splenic autotransplantation combined with lower esophagus transaction anastomosis is a safe and effective treatment strategy for patients with liver cirrhosis induced portal hypertension, and the spleen tissue transplanted into the retroperitoneal space can partially preserve the immune function. PMID- 19562998 TI - [Nonoperative management of blunt splenic injury]. AB - OBJECTIVE: To investigate the monitoring and therapeutic methods of nonoperative management of blunt splenic injury. METHODS: Eighty-two cases with nonoperative management of 95 patients of blunt splenic injury from September 2005 to April 2008 were analyzed retrospectively. Percutaneous peritoneal drainage was applied to 75 cases, and auto-blood transfusion was applied to 38 cases. Eighty-two cases were followed up from 3 weeks to 8 months. RESULTS: Eighty-two patients with nonoperative management were treated successfully, including 34 cases classified as grade III to IV, 6 cases over 55-years-old, 14 cases with severe multiple injury (ISS > or = 16) and 37 cases whose drained peritoneal blood volume were over 500 ml. The drained peritoneal blood volume was 30 to 2400 ml. The total volume of auto-blood transfusion was 22 300 ml and the average volume was 613 ml. All cases were followed up without delayed hematocele or peritoneal infection. CONCLUSIONS: Most hemodynamically stable patients with blunt splenic injury can be healed with nonoperative management. The treatments including percutaneous peritoneal drainage and transfusion of auto-blood can significantly increase the performance rate and the achievement ratio of nonoperative management of blunt splenic injury. PMID- 19562999 TI - [Changes in splenic macrophage function of hypersplenism due to portal hypertension]. AB - OBJECTIVE: To investigate the changes in function of splenic macrophages of hypersplenism due to portal hypertension (PH), and to provide experimental evidence for exploring the immune function of spleen in PH. METHODS: Twelve patients with hypersplenism due to PH and four patients with traumatic rupture of spleen, from September 2005 to March 2006, were enrolled into PH group and control group, respectively. Splenic M phi were isolated and purified by anchoring cultivation from all the patients, and were resuspended by RPMI-1640. Phagocytosis, cytokine secretion and antigen processing and presenting of splenic M phi were detected by Vybrant Phagocytosis Assay, the human TNF-alpha Elispot kits and DQ ovalbumin. RESULTS: Compare to the normal splenic M phi, the phagocytosis rate, antigen presentation positive cells and secretion positive cells, were all significantly increased in PH splenic M phi (86.4 +/- 7.1 vs. 61.8 +/- 4.1, 26.3 +/- 1.6 vs. 15.6 +/- 1.8, 387.0 +/- 24.3 vs. 240.3 +/- 13.0, P<0.01). CONCLUSIONS: The phagocytosis, cytokine secretion and antigen processing and presenting of splenic M phi in PH spleen were all significantly increased, and the M phi retained at activated state. It means that the PH spleen still possessed the immune function, but these functions might be in disorder. It still needs more research to get the precious evaluation for immune function in the PH spleen. PMID- 19563000 TI - [Analysis of the surgical outcome and prognostic factors for hilar cholangiocarcinoma]. AB - OBJECTIVE: To assess the therapeutic strategies and prognostic factors which influence on clinical outcome of hilar cholangiocarcinoma. METHODS: A total of 144 patients with hilar cholangiocarcinoma underwent operation between January 1990 and December 2005 were analyzed, including 102 males and 42 females with 36- 74-years-old. All patients underwent resection among which 86 cases (59.7%) had an R0 resection (negative histologic margins), 34 cases (23.6%) had an R1 resection (positive histologic margins), 24 cases (16.7%) had an R2 resection. The Bismuth-Corlette classification of group R0 and R1: 28 cases (23.3%) in type I , 49 cases (40.8%) in type II, 10 cases (8.3%) in type III A, 19 cases (15. 8%) in type III B and 14 cases (11.7%) in type IV. The TNM stages of group R0 and R1: 19 cases (15.8%) in stage I, 80 cases in stage II (66.7%), 16 cases in stage III (13.3%), 5 cases in stage IV (4.2%). In group R0 and R1, there were 41 cases with well differentiated and 79 cases with moderately and poorly differentiated, 62 cases (51.7%) with negative lymph nodes and 58 cases (48.3%) with positive lymph nodes, 42 cases in stage T1 and 78 cases in stage T2-3, 86 cases with negative blood vessel metastasis and 34 cases with positive blood vessel metastasis. RESULTS: The median survival time was 46.8 months after R0 resection, 18.3 months after R1 resection, and 11.2 months after R2 resection. The 1-, 3- and 5-year cumulative survival rates of the patients were 60.2%, 36.1% and 29.4%. Survival rates after resection in patients with negative lymph nodes (n = 62) were significantly longer than that in those with positive lymph nodes (n = 58) (P < 0.01). The T stage system predicted respectability and the likelihood of an R0 resection and correlated with survival (P = 0.030). Patients requiring portal vein resection had a worse prognosis than those without vascular resection (P = 0.047) but still survived longer than patients who were unresectable (P < 0.01). CONCLUSIONS: Negative histologic margins, concomitant partial hepatectomy, and well-differentiated tumor histology are associated with improved outcome after all hilar cholangiocarcinoma resections. In patients who underwent an R0 resection, concomitant partial hepatectomy is the only independent predictor of long-term survival. PMID- 19563001 TI - [Surgical treatment for primary gastrointestinal stromal tumors: a report of 73 cases]. AB - OBJECTIVE: To investigate the effects and prognosis of surgical treatment in primary gastrointestinal stromal tumors (GIST). METHODS: The clinicopathological data of 73 patients with primary GIST underwent operation from April 1997 to December 2007 was retrospectively analyzed, and the prognosis was evaluated too. RESULTS: Among the 73 cases, 68 cases received complete tumor resection, among which 12 cases underwent laparoscopic operation; while palliative resection and biopsy only were carried out in the other 5 cases. There was significant difference in survival rate between the two groups (P = 0.000). The 1-, 3-, 5 year survival rates of the 66 cases had been followed up was 91.0%, 78.2% and 74.1%, respectively. The malignancy risk grades of GIST was related to the survival rates on statistical analysis (P = 0.002). Significant differences were found in the survival rates between the patients with very low grade, low grade and high grade malignancy tumors (P = 0.012, 0.002). CONCLUSIONS: Complete tumor resection should be emphasized in primary GIST, and more attention should be paid to the initial surgical treatment. Extended surgical resection is required for tumors of higher malignancy risk. The indications of laparoscopic surgery in GIST should be selected with caution for tumor complete resection. PMID- 19563002 TI - [Time pattern and prognostic evaluation of the recurrence of rectal cancer after resection]. AB - OBJECTIVE: To investigate the regular pattern and prognostic evaluation of patients with recurrent rectal cancer after resection. METHODS: Three hundred and fourteen cases with recurrent rectal cancer after resection treated between May 1979 and November 2006 were classified into three groups according to the recurrence intervals (<3 years, 3-5 years, >5 years). The survival rates and prognosis in the three groups were analyzed and compared retrospectively. RESULTS: Of the 314 patients, the cancer relapsed in 247 cases (78.7%) in less than 3 years, and the recurrence occurred predominantly within 2 years (207 cases, 65.9%) after the operation. The neoplasm in 41 cases (13.3%) recurred in 3 5 years after the operation, and 26 cases (8. 3%) in more than 5 years after the resection. Disease-free interval, Dukes stage, neoplasm gross type, histological type, T stage, lymphatic and distant metastasis were associated with the prognosis on univariate analysis. And disease-free interval and tumor Dukes stage were independent prognostic factors for survival rate on multivariate analysis. Disease-free interval and progression-free survival were related positively with survival time. CONCLUSIONS: The rectal cancer patients should be followed-up intensively for 2 years after the operation and moderately from then on. Disease free interval and progression-free survival could be taken as the best predictors of long-term cure and prognosis. PMID- 19563003 TI - [Expression features of P-glycoprotein, glutathione S transferase-pi and inhibitor of apoptosis proteins in lymph node metastases of gastrointestinal carcinomas]. AB - OBJECTIVE: To investigate the expression features of P-glycoprotein (P-gp), glutathione S transferase-pi (GST-pi) and inhibitor of apoptosis proteins like p53, survivin and bcl-2 in lymph node metastases of gastrointestinal carcinomas. METHODS: The expression of P-gp, GST-pi, p53, survivin and bcl-2 were determined by using immunohistochemistry technique in surgical specimens of primary tumor (PT) and lymph node metastases (LNMs) from 54 gastrointestinal cancer patients with metastasis of lymph nodes. The expression difference of 5 multi-drug resistance (MDR)-related factors between LNMs and PT were compared. RESULTS: Significant difference was found in the expression of P-gp and GST-pi between the two groups (both P < 0.05), and expression of p53 and bcl-2 showed positive correlation between LNMs and PT (r = 0.7248, 0.5524; both P < 0.05), respectively. In LNMs, P-gp expression was positively correlated with GST-pi (r = 0.4062, P < 0.05) and survivin (r = 0.6169, P < 0.05), and also GST-pi expression was related positively with survivin (r = 0.4027, P < 0.05). Statistically positive correlations were noted between bcl-2 and P-gp (r = 0.3986, P < 0.05), bcl-2 and survivin (r = 0.2937, P < 0.05), as well as GST-pi and survivin (r = 0.4481, P < 0.01) in PT. Only a positive correlation between GST-pi and survivin expression was simultaneously shown in both LNMs and PT. CONCLUSIONS: There is significant heterogeneity of MDR-related factors expression in LNMs of gastrointestinal carcinomas. Effective adjuvant chemotherapy after operation should target on the metastatic loci of the disease. PMID- 19563004 TI - [Adult hematogenous pyogenic discitis and vertebral osteomyelitis: clinical features, diagnostic, therapeutic approaches and outcome in 23 patients]. AB - OBJECTIVE: In order to sum up the experience of diagnosis and treatment, the clinical information, diagnostic approach and treatment strategies from 23 patients with hematogenous pyogenic discitis and vertebral osteomyelitis (PDVO) in adults were reviewed. METHODS: A retrospective record review was conducted of all cases of hematogenous PDVO from July 1999 to October 2006, the diagnostic approach and the treatment strategies were discussed. RESULTS: The average age was 55.4 years. The average delayed diagnosis time was 4.4 months (range, 4 weeks to 11 months). Of these, 17 had underlying diseases such as diabetes mellitus, chronic alcoholism, liver cirrhosis, rheumatic disease, and the use of corticosteroids. All percent had elevated erythrocyte sedimentation rates and C reactive proteins, while white blood cell counts were less reliably elevated. Imaging studies included radiographs and MRI. Seven patients were treated with immobilization and intravenous antibiotic drugs. Because of a delay in diagnosis, 16 patients underwent operative treatment for hematogenous PDVO using staged anterior debridement and Bone grafting with iliac crest or rib strut, anterior spinal instrumentation in 3 patients, posterior instrumentation was placed in 7 patients at a second procedure 10 days to 2 weeks following initial operation. Twenty-three patients were followed-up for an average period of 27 months, with a minimum of 6 months and the longest for 7 years. The patients return to their preoperative everyday activity. No patient had a recurrence of osteomyelitis. All their symptoms improved after surgery, bony fusion occurred in 87.5% of cases. CONCLUSIONS: Adult hematogenous PDVO is a disease that affects mainly older patients suffering underlying medical illnesses. Positive blood cultures is valuable for the diagnosis. Hematogenous PDVO may require surgery in case of a development of biomechanical instability and/or a vertebral collapse with progressive deformity. In experienced hands, surgical debridement, interbody fusion, and anterior or posterior instrumentation is a safe and effective treatment. PMID- 19563005 TI - [Preliminary experiences in minimally invasive transforaminal lumbar interbody fusion]. AB - OBJECTIVE: To explore clinical application of minimally invasive transforaminal lumbar interbody fusion (TLIF) for the management of lumbar disorders and discuss its indications, surgical techniques and clinical effectiveness. METHODS: From Jan 2005 to Dec 2006, 31 selected patients (22 males and 9 females, aged from 41 to 63 years) with degenerative lumbar diseases were treated with minimally invasive TLIF assisted by METRx X-Tube micro-endoscopy system. The index diagnosis was lumbar disc herniation with Lumbar spinal stenosis in 7, lumbar disc herniation with segmental instability in 16, grade 1 to 2 of lumbar spondylolisthesis in 8. The surgical methods was performed with bilateral or unilateral pedicle screws insertion and a single rectangle cage posterolateral placement. The operating time, blood loss, blood transfusion, drainage, visual analogue scale (VAS), preoperative and postoperative JOA scores were observed as well as radiographic evaluation. The results were compared with standard TLIF group respectively. RESULTS: A total of 116 pedicle screws and 31 cages were implanted of which 4 patients were treated with unilateral pedicle screws fixation. Four pedicle screws were found misplaced in CT scans after surgery. The average operating time was 199 min, blood loss 359 ml, volume of transfusion 32 ml drainage 81 ml, and VAS was 2.37 about 72 hours after surgery, which had statistic difference compared with control group. There was no statistic difference on postoperative improvement rate and JOA scores in two groups. CONCLUSIONS: Minimally invasive TLIF minimizes paraspinal muscle trauma and blood loss, shortens the operating and recovery time. A good long-term outcome can be gained compared with standard procedures. PMID- 19563006 TI - [Effects of perioperative administration of celecoxib on pain management and recovery of function after total knee replacement]. AB - OBJECTIVE: To assess the effect of perioperative administration of a selective cyclooxygenase 2 inhibitor (celecoxib) on pain management and recovery of function after total knee arthroplasty (TKA). METHODS: Randomized, controlled trial conducted from January 2005 through February 2006, 60 patients underwent TKA for osteoarthritis or rheumatoid arthritis were randomly divided into group of perioperative, administration of celecoxib (Study group, n = 30) and postoperative administration of celecoxib (Control group, n = 30). Patients in Study group were given oral celecoxib 3 d before TKA, 200 mg twice daily, and extended to 5 d postoperatively; patients in Control group were given oral celecoxib 2 h after TKA, 200 mg twice daily, and extended to 5 d postoperatively. All operations were finished by the same surgeon group. RESULTS: The postoperative patient-controlled analgesia (PCA) consumption was significantly less in Study group than in Control group [(43 +/- 12) ml vs. (53 +/- 12) ml, P < 0.05]. The pain scores of postoperative 4, 8, 12 h, 1, 2 d in Study group were 6.1 +/- 1.2, 5.0 +/- 1.3, 4.3 +/- 1.1, 3.4 +/- 1.2, significantly less than in Control group (P < 0.05); There were no intergroup significant differences in the pain scores of postoperative 3, 4, 5 d (P > 0.05). There were no intergroup significant differences in respect to the side-effect occurrence, operation time and postoperative drainage, postoperative analgesic consumption (P > 0.05). The time to achieve 90 degrees knee flexion was significantly shorter in Study group than in Control group [(6.2 +/- 1.7) d vs. (8.6 +/- 1.8) d, P < 0.05]. CONCLUSIONS: Perioperative administration of the selective Celecoxib holds the effect of preemptive analgesia. Compared with postoperative administration, perioperative administration of celecoxib can alleviate the early postoperative pain score, reduce the consumption of postoperative analgesic, accelerate the recovery of joint motion and thus increase the patient satisfaction. PMID- 19563007 TI - [Survival and prognostic factors in resected satellite-nodule T4 non-small cell lung cancer]. AB - OBJECTIVE: To study the survival and prognostic implication in surgically resected satellite-nodule T4 (T4 satellite) non-small cell lung cancer (NSCLC). METHODS: From January 1995 to March 2005, the complete resection was performed to 42 patients with NSCLC who were postoperatively identified as pathologic-stage T4 satellite. Survival and associations between clinicopathological parameters and prognosis were analyzed. Thirty-two patients with pathologic stage local-invasion T4 (T4 invasion) NSCLC who underwent resection at the same time were also analyzed. RESULTS: The 1-, 3- and 5-year survival was 76.2%, 57.1% and 46.0% for patients with T4 satellite, while 62.3%, 31.5% and 20.0% for patients with T4 invasion. There was a significant higher survival in T4 satellite group when compared to that in T4 invasion group (P < 0.05). Furthermore, patients with T4 satellite N0M0 got a better survival than those with T4 satellite N1-2M0, T4 invasion N0M0 and T4 invasion N1 -2M0 (P < 0.05). For patients with T4 satellite, univariate analysis showed that histology, main tumor size, lymph node status and adjuvant chemotherapy were linked with survival, while main tumor size, lymph node status and adjuvant chemotherapy served as the independent prognostic factors with multivariate analysis. CONCLUSIONS: Patients with completely resected T4 satellite NSCLC have a better prognosis than those with T4 invasion. Main tumor size over 3 cm, lymph node metastasis or no adjuvant chemotherapy means an unfavorable prognosis. PMID- 19563008 TI - [Treatment strategy of pituitary invasive prolactinomas]. AB - OBJECTIVE: To discuss the treatment strategy of invasive prolactinomas (IPs) involving the cavernous sinus. METHODS: Data from 80 patients with IPs treated in our institutions were reviewed retrospectively. The criteria utilized included: (1) invasion of the cavernous sinus by tumor, corresponding to Grade III-IV according to the classification of Knosp; (2) serum prolactin level > 9.1 nmol/L; (3) clinical signs of hyperprolactinemia and mass effect. Among the 80 patients who met the criteria: 21 patients received bromocriptine as primary treatment (Group A); 21 patients initially received bromocriptine and then accepted microsurgery or irradiation (Group B); 38 patients had initially undergone transcranial or transsphenoidal microsurgery and then received bromocriptine or adjuvant radiotherapy (Group C). Eleven patients underwent gamma knife radiotherapy. RESULTS: In 57 patients (12 cases of Group A, 16 cases of Group B, 29 cases of Group C), the tumors on MRI had almost completely disappeared after an average follow-up period of 62 months, and in the other 23 patients, residual tumor involved the cavernous sinus. Visual symptoms improved in 33 patients while deteriorated in 7 patients. Serum prolactin level of 52 patients had in normal range after treatment (10 cases of Group A, 11 cases of Group B, 31 cases of Group C) and 7 patients were more than 9.1 nmol/L. Nine patients had symptoms of hypopituitarism. CONCLUSIONS: For IPs, individualized treatment methods are advocated in which dopamine agonist medications are effective as first-line therapy. It is necessary to take dopamine agonist after operation and close observation is mandatory. Gamma knife surgery is an option to treat the residual tumor involving the cavernous sinus. PMID- 19563009 TI - [Neuromodulation for treatment for neurogenic bowel dysfunction]. AB - OBJECTIVE: To explore the efficacy of neuromodulation (including sacral neuromodulation and dorsal penile/clitoral nerve neuromodulation) for the treatment to neurogenic bowel dysfunction due to spinal cord injury. METHODS: From January 2006 to April 2008, 9 patients with neurogenic constipation after spinal cord injury underwent the therapy of neuromodulation, 1 patient underwent the therapy of sacral neuromodulation, 8 patients underwent the therapy of dorsal penile/clitoral nerve neuromodulation. The therapeutic efficacy was evaluated and followed up by means of Wexner constipation score. RESULTS: One patient received permanent electrode and neurostimulator implantation and constipation were improved continuously. A significant improvement in the Wexner constipation score was observed compared with the preoperative baseline level (preoperative baseline: median 22; after implantation: median 9). Four patients were effective after the therapy of dorsal penile/clitoral nerve neuromodulation. Wexner constipation score decrease from 19 to 11 after 12 weeks dorsal penile/clitoral nerve neuromodulation. Patients also showed a significant improvement in their symptoms and quality of life during follow up. CONCLUSIONS: Sacral neuromodulation and dorsal penile/clitoral nerve neuromodulation may be effective for some neurogenic constipation. However there are no methods successfully identify the candidate who will be beneficial before the procedure. Good quality research data are needed to evaluate the effects of sacral neuromodulation and dorsal penile/clitoral nerve neuromodulation for these conditions. PMID- 19563010 TI - [Three-D gait patterns before and after total hip arthroplasty]. AB - OBJECTIVE: To determine how selected gait parameters may change as a result of total hip arthroplasty (THA). METHODS: From February 2006 to February 2007 the study was performed on 53 osteonecrosis of femoral head subjects prior to and 6 weeks, 3, 6 and 12 months after total hip arthroplasty surgery. Gait analysis was performed using the three-dimensional Infrared-based system (Dalian Dong Fang Xin Rui Company, China). Kinematic data were recorded for the lower limb. The results obtained from the osteonecrosis of femoral head subjects were compared with those of 40 individuals without osteonecrosis. Using a three-dimensional motion analyzer, the following were measured: (1) gait speed, stride length, cadence; (2) step length, stride width; (3) standing phase time, walking phase time; (4) gait cycle time, Joint movement angular. RESULTS: Compared with persons before total hip arthroplasty, THA patients showed significantly increased gait speed, gait frequency, step length and affected limb stand phase (P < 0.01). Angle timetable: preoperative hip-maximum flexion was 15 degrees, post-extension was 8 degrees, knee-maximum flexion was 70 degrees, ankle-maximum flexion was 15 degrees. Postop hip-maximum flexion was 27, post-extension was 17 degrees, knee maximum flexion was 50 degrees, ankle-maximum flexion was 14 degrees; parameters of hip, knee between preoperative and postoperative was significance (P < 0.01). CONCLUSION: These results suggest that 3-D gait analysis before and after total hip arthroplasty can evaluate precisely hip vitodynamics variation. PMID- 19563011 TI - [An experimental study of correction of idiopathic-type scoliosis by staple]. AB - OBJECTIVE: To evaluate the feasibility of the correction idiopathic-type scoliosis by implanting the staple in growing animal models. METHODS: Fourteen female goats were performed unilateral pedicle screws asymmetric tethering in left side in combination with right rib resection (age: 5 to 8 weeks, weight: 6 to 8 kg). The observing time was about 8 weeks. Goats that had been created scoliosis model successfully were classified in 2 groups randomly. CONTROL GROUP: just removing the posterior tether, no treatment was offered. Correct group: the removing of posterior tether and the stapling of anterior spinal epiphysis were performed simultaneously. Dorsoventral and lateral plain radiographs were taken preoperatively and postoperatively. Serial X-ray postoperatively were performed every 4 weeks to measure the Cobb angle of the spine and to observe the condition of the insert. The observing time is about 8 weeks. RESULTS: Radiography showed that 12 goats had created scoliosis model successfully. CONTROL GROUP (n = 6): Series X-ray show that the change of the Cobb angle was not obviously. The initial curves after the procedures measured an average of 40.8 degrees (28 degrees-56 degrees), the average Cobb angle was 42.5 degrees (30 degrees-58 degrees) after 8 weeks, no statistics difference are found (P > 0.05). Treatment group (n = 6): no complication such as pedicel screw break, instrument loosen, dislocation, injury of blood vessel, nerve injury and organ injury of thoracic cavity etc, were found during the observing period. The initial curves after the procedures measured an average of 44.5 degrees (36 degrees-57 degrees), to some degree, the Cobb angle decreased and the average was 42.5 degrees (30 degrees-58 degrees) after 8 weeks. There are statistics difference between the initial and final curves (P < 0.05). CONCLUSION: As a means of mechanical modulation, stapling can be manipulate conveniently and safely, and can modulate the spinal growth of the animal model successfully, predicted that it may be a new selection for idiopathic-type scoliosis in growing children. PMID- 19563012 TI - [Binding pancreaticogastrostomy]. AB - OBJECTIVE: To discuss the value of a new technique of the binding pancreaticogastrostomy (BPG) in pancreaticoduodenectomy. METHODS: From May 2008 to October 2008, 15 patients were performed with BPG, included pancreatic head cancer in 7 cases, duodenal adenocarcinoma in 2 cases,mass-type chronic pancreatitis with pancreatolithiasis in 1 case, ampullary carcinoma in 1 case, gallbladder cancer in 1 case, islet cell tumor in 1 case and cholangiocarcinoma in 2 cases. The main procedures of BPG included: isolating remnant pancreas; slitting partial posterior wall of stomach and preplaced with seromuscular purse string suture; cutting gastric anterior wall; performing pancreaticogastrostomy (binding of outer seromuscular and inner mucous layer of stomach). RESULTS: The procedures were successful in 15 patients. Postoperative complications included small amount of pleural effusion in 2 cases, delayed gastric emptying in 2 cases and bile leakage in 2 cases. All patients were cured in 2 weeks. No mortality and anastomosis leakage occurred. CONCLUSION: The application of BPG technique can prevent the anastomosis leakage and improve the safety for pancreaticoduodenectomy. PMID- 19563013 TI - [Initial experience of partial cystectomy in treatment of bladder tumor by 2 microm continuous wave laser]. AB - OBJECTIVE: To analyze the clinical characteristics of partial cystectomy in treatment of bladder tumor by 2 microm continuous wave laser. METHODS: A total of 18 patients with 21 bladder tumors treated by 2 microm laser via transurethral under sacral block. The diameters of tumors were between 1-3 cm. In the operation, we vaporized and cut whole wall of bladder, dissected the muscular layers from connective tissue, and sent them for pathology inspection. We reviewed these following variables: operation time, blood loss, complications during and after the operation, pathology staging of the tumor and follow-up visit. RESULTS: All the patients were tolerated in the operation. The mean operative time was (7.4 +/- 3.3) min (range, 5-12 min). Blood loss in the operation was minimal. No patient experienced an obturator nerve reflection or a hemorrhage after the operation. One case has urine leakage. The pathology staging of the tumors were as follows: T1 stage 15 cases, total 18 tumors; and T2 stage 3 cases, total 3 tumors. The followup periods were between 3 to 6 months, and the mean period was 4.5 months. There was no recurrence in situ. CONCLUSIONS: The 2 microm laser not only can vaporize and cut the wall of bladder finely, but also can dissect the muscular layers from connective tissue efficiently. So it can be partial cystectomy in the treatment of bladder tumor. PMID- 19563014 TI - [Clinical study of allogeneic hematopoietic stem cell transplantation for relapsed/refractory acute lymphocytic leukemia]. AB - OBJECTIVE: To explore the efficacy and toxicity of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for relapsed/refractory acute lymphocytic leukemia (ALL). METHODS: Forty-seven patients with relapsed/refractory ALL received allo-HSCT, which containing 19/47 from HLA-identical sibling donors (sib HSCT), 18/47 from HLA-identical unrelated donors (URD-HSCT) and 10/47 from haplo identical donors (Hi-HSCT). Conditioning regimens included "TBI plus Cyclophosphamide (Cy) (42/ 47)" or "busulfan (Bu) plus Cy (5/47)". Cyclosporine (CsA) combined with a short-course Methotrexate (MTX) were used for graft versus host disease (GVHD) prophylaxis. In addition, patients receiving URD-HSCT or Hi HSCT were given mycophenolate mofetil (MMF) and anti-thymocyte immunoglobulin (ATG). Patients with molecular or cytogenetic relapse tendency on minimal residual disease (MRD) monitoring received donor lymphocyte infusion (DLI). RESULTS: All patients tolerated the therapy well except for mucositis. Renal dysfunction occurred in 2 patients on CsA therapy. Epilepsy occurred in 1 patient, fatal infectious complications in 9 (including 3 interstitial pneumonia), grade III-IV acute GVHD (aGVHD) in 7, chronic GVHD (cGVHD) in 22 and hemorrhagic cystitis (HC) in 4 patients. Thirteen patients relapsed after transplantation. The median time of hematopoietic reconstitution was + 17 ds. Nineteen patients received DLI, and 6 of them had no disease progression. With a median follow-up duration of 43 (10-77) months, the estimated 5-year overall survival (OS) and disease free survival (DFS) rates were 49.65% and 46.55%, respectively. CONCLUSION: Allo-HSCT is an effective therapy for relapsed/refractory ALL. Relapse after transplantation, fatal infection, and severe acute GVHD are the main causes for failure. DLI might decrease the relapse rate after transplantation. PMID- 19563015 TI - [Application of real time polymerase chain reaction to the diagnosis and treatment of cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation]. AB - OBJECTIVE: To investigate the role of real time quantitative polymerase chain reaction (RQ-PCR) in the diagnosis and treatment of recipients cytomegalovirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (allo HSCT). METHODS: 318 patients received allo-HSCT were studied. 160 patients received transplants from HLA matched sibling donors; 127 from HLA mismatched related donors; 31 from unrelated donors. Before transplant recipients and donors received CMV serological test by ELISA. After transplant RQ-PCR was used to test and monitor CMV-DNA in plasma of patients. A positive CMV-PCR was defined as > 6 x 10(2) copies/ml. Ganciclovir was used for CMV prophylaxis in all patients at -9 d to -2 d of conditioning regimen period. Ganciclovir, foscarnet, or combination of the two drugs were used as the preemptive therapy. RESULTS: The total 100-day cumulative incidence of CMV infection was 40.6%. The incidence was 17.5%, 66.1% and 45.2% for the HLA matched sibling, HLA mismatched related (MMR) and unrelated donor (MUR) HSCT respectively. Multivariate analysis showed MMR HSCT, MUR HSCT, ATG containing preparative regimen and moderate to severe aGVHD were the risk factors for CMV infection after HSCT. The 100 day cumulative incidence of CMV disease was 8.8% and 5.6%, 9.4%, 22.6% respectively for total and three kinds of HSCT after early preemptive therapy. Two-year survival of CMV infection was similar in the three kinds of SCT. CONCLUSION: Detection of CMV DNA in plasma by real time PCR appears to be effective for the diagnosis and surveillance of CMV infection after HSCT. It may help to initiate antiviral therapy and reduce the incidence of CMV disease in the patients with high risk of CMV infection. PMID- 19563016 TI - [Non-T-cell depleted HLA haploidentical peripheral blood stem cell transplantation for hematological malignancies: report of 36 cases]. AB - OBJECTIVE: To analyze the clinical outcome of human leukocyte antigen (HLA) haploidentical peripheral blood stem cell transplantation (PBSCT) from related donors for hematological malignancies. METHODS: Thirty-six patients with hematological malignancies, with a median age of 25 (11-48) years, were transplanted with PBSC from an HLA-haploidentical family donors: 7 were 1 locus mismatched and 29 were 2-3 loci mismatched. The recipients received myeloablative conditioning regimen, in combination with different immunosuppressants according to the degree of HLA disparity followed by non-T-cell depleted PBSCT. The median number of CD34+ cells were 11 (4.16-21.00) x 10(6)/kg. RESULTS: All patients achieved sustained, full donor-type engraftment. Fifteen patients (41.7%) developed grade I-II aGVHD. Among 29 patients followed up more than 18 months, 17 (58.6%) developed cGVHD. There was no statistical difference in decrease and recovery of T, B and NK cell subsets after transplantation between HLA haploidentical group and HLA identical PBSCT group. The median follow-up duration was 15 (4 -69) months. Five patients (13.9% ) relapsed. The 2-year probability of leukemia-free survival (LFS) was 82.2%. CONCLUSION: Non-T-cell depleted HLA haploidentical PBSCT is safe and feasible for patients with hematological malignancies after myeloablative conditioning regimen combined with intensive immunosuppressants. PMID- 19563017 TI - [Influence of methylprednisolone on cell component of donor graft and on H-2 haploidentical hematopoietic stem cell transplantation in mice]. AB - OBJECTIVE: To explore the influence of methylprednisolone (MP) on cellular component in donor graft and on H-2 haploidentical hematopoietic stem cell transplantation (HSCT) in mice. METHODS: A murine model of H-2 haploidentical HSCT was established by using of c57BL/6J male mouse as donor and (c57BL/6J x LB/C) F1 female mouse as recipient. The donor mouse received peripheral-blood (PB) progenitor cells mobilization regimens consisted of recombinant human granulocyte colony-stimulating factor (rhGCSF) alone (control group) or combined with MP in dose of 2 mg/kg daily [small-dose (SD) group], 10 mg/kg daily [middle dose (MD) group], and 50 mg/kg daily [large-dose (LD) group] respectively. Percentage of T cell subsets, DC1 (HLA-DR+CD11c+) and CD34+ cell in the grafts were detected by flow cytometry. Transplant rejection,severity of GVHD and survival time were observed. RESULTS: The percentages of CD3+ T cell in donor grafts in the three groups were significantly lower than that in control group (P < 0.05). The percentage of CD3+ CD4+ T cells decreased more significantly than that of CD3+ CD8+ T cells, and CD4/CD8 ratios decreased significantly. The percentage of CD4+ CD25+ T cells increased significantly, the percentage of DC1( HLA-DR+CD11c+) decreased and the percentage of CD34+ cells increased in all the three groups than in control group. There were significant differences in the percentage of CD3+ T cells, CD3+ CD4+ T cells and CD34+ cells in donor grafts among SD group, MD group and LD group (P < 0.05). The engraftment rates in control, SD, MD and LD groups were 90%, 100%, 100% and 80% respectively. Severity of aGVHD in each study group decreased significantly compared with that in control group (P < 0.05). There were statistical differences among different dosage groups (P < 0.05). Survival time after transplantation in all study groups were significantly longer than that in control group (P < 0.05), and in MD group was significantly longer than in SD group and LD groups (P < 0.05). CONCLUSIONS: Addition of methylprednisolone to routine donor mice HSC mobilization regimen has a definite effect in alleviating aGVHD and prolonging survival time of mouse after H-2 haploidentical HSCT. With a suitable dosage addition of methylprednisolone to donor mice HSC mobilization regimen could avoid the increasing risk of graft rejection. PMID- 19563018 TI - [Study on allelic characteristics of "homozygote" resulted from low resolution genotyping of HLA-Cw locus]. AB - OBJECTIVE: To study the allelic characteristics of "homozygote" resulted from low resolution genotyping of HLA-Cw locus and to provide more precise typing data for clinical transplantation. METHODS: Forty three related allogeneic hematopoietic stem cell transplantation(allo-HSCT) donors and patients with HLA-Cw * 03, Cw * 07 homozygote, which were the most common gene groups in Chinese population, identified by low resolution genotyping level were retyped by high resolution PCR SSP genotyping method, and three dimensional structure modelling was also made by using a solely developed HLA three-dimensional matching software (HLA strucMark version 1.0) to evaluate the effect of differences between two mismatched alleles and its relationship with GVHD occurrence. RESULTS: The typing results of high resolution level demonstrated that the confirmed allelic homozygotes for Cw * 03 and Cw * 07 were 14% and 43%, respectively, while the others were all heterozygotes. The ambiguous typing results could be confirmed by family data study or by high resolution typing methods when there was no family data available, Three-dimensional modeling results of the mismatched alleles undetected in low resolution typing level indicated that family data study or high resolution PCR-SSP genotyping were important in preventing graft-versus-host disease. CONCLUSIONS: When HLA-Cw homozygote results were found in low resolution genotyping method, the results should be reconfirmed by family data study or by high resolution typing methods to provide precise typing results for avoiding severe graft-versus-host disease. PMID- 19563019 TI - [Study on the mechanism of enhancing homing efficiency of human hematopoietic stem/progenitor cells into bone marrow after manipulation with tumor necrosis factor alpha in xenotransplanted BALB/c mouse model]. AB - OBJECTIVE: To investigate the role of tumor necrosis factor (TNF) alpha on the homing efficiency of hematopoietic stem/progenitor cells (HS/PC) into bone marrow and its mechanism. METHODS: CFSE-labeled umbilical cord blood (UCB) CD34+ cells were transplanted into irradiated (control group) or combined with TNF alpha prepared (experimental group) BALB/c recipient mice. The distribution in peripheral blood, liver, lung and homing characteristics in bone marrow and spleen of UCB CD34+ cells, in BALB/c recipient mice were determined 20 hours after xenotransplantation by flow cytometry (FACS) and their homing efficiency was calculated. ELISA was used to measure serum SDF-1 alpha level. CXCR4 expression levels of on UCB CD34+ cells were assessed by FACS pre-/post manipulation with TNF alpha. SDF-1 alpha expression level in bone marrow and spleen was tested by immunohistochemistry. RESULTS: UCB CD34+ cells mainly home into recipient mice bone marrow and spleen; The homing efficiency in experimental group bone marrow [(0.65 +/- 0.13)%] was significantly higher than that in control ones [(0.30 +/- 0.09)%, P < 0.01], whereas the homing efficiency in experimental group spleen was dramatically lower than that in control ones (P < 0.01); Treatment with TNF alpha did not affect recipient serum SDF-1 alpha level; After 18 hours co-cultured with TNF alpha, the CXCR4e expression level on UCB CD34+ cells was similar to that on fresh ones; TNF alpha treatment induced significantly higher SDF-1 alpha expression on osteoblastic and stromal cells in bone marrow, and reversed spleen SDF-1 alpha gradient that was originally favorable for CD34+ cells homing. CONCLUSION: TNF alpha enhances the homing efficiency of HS/PC via up-regulating SDF-1 alpha gradient in bone marrow, and might be an useful enhancer for HS/PC homing in clinical practice. PMID- 19563020 TI - [Study of influence of umbilical cord mesenchymal stem cells on CD34+ cells in vivo homing in NOD/SCID]. AB - OBJECTIVE: To investigate the effect and the potential mechanism of umbilical cord (UC) derived mesenchymal stem cells (MSCs) on umbilical cord blood (UCB) derived CD34+ cells in vivo homing in xenotransplanted NOD/SCID mice model. METHODS: CD34+ cells and MSCs were derived from fresh UCB and UC, respectively. CD34+ cells (5 x 10(5) per mice) and MSC cells (5 x 10(6) per mice) were co transplanted into irradiated NOD/SCID mice intravenously. CD34+ cells (5 x 10(5) per mice) alone were transplanted into the mice as control group. CD34+ cells home in bone marrow and spleen of recipient mice were detected 20 hours after transplant by FACS and RT-PCR, and the homing efficiencies were calculated. The effect of MSCs on CD34+ cells chemotactic function was investigated after co cultured UCB CD34+ cells with UC MSCs in vitro. After 4 and 7 days coculture, the homing related adhesion molecules (the CD49e, CD31, CD62L, CD11a) expressed on CD34+ cells were detected by FACS. RESULTS: 1) The homing efficiencies in bone marrow in experimental and control group were (7.2 +/- 1.1)% and (5.4 +/- 0.9)%, respectively (P < 0.05). 2) Human GAPDH gene was detected in bone marrow in experimental group and in spleen in both groups. 3) The migration efficiency of CD34+ cells was significantly higher in experimental group (35.7 +/- 5.8)% than in control group (3.5 +/- 0.6)% (P < 0.05). 4) The expression of CD49e, CD31, CD62L on CD34+ cells kept higher level in MSCs cocultured group than in CD34+ cells alone group. CONCLUSIONS: MSCs can efficiently increase homing of CD34+ cells to bone marrow and spleen in vivo by keeping a high level of homing adhesion molecules expression and improving migration efficiency of UCB CD34+ cells. PMID- 19563021 TI - [The action of donor-derived NK cell in leukemic mice MHC haplotype-mismatched bone marrow transplantation]. AB - OBJECTIVE: To study the role of donor-derived NK cells in haploidentical bone marrow transplantation (BMT) in leukemic mice. METHODS: CB6F1(H-2b/d) mice model of EL9611 (H-2d) erythroleukemia was established by injection of EL9611 (H-2d) cells via tail vein. CB6F1(H-2b/d) mice were used as recipient, and C57BL/6(H-2b) mice as donor. Five days later, 70 CB6F1(H-2b/d) mice were randomly divided into 7 groups (10 mice per group) as follows: group 1: without treatment; group 2: simple-irradiated group; group 3: treated with cytarabine (Ara-C) at 50 mg/kg x6 d; group 4: simple BMT; group 5: haploidentical BMT with graft-versus-host disease (GVHD) that injected with bone marrow cells and spleen cells of C57BL/6(H 2b) mice 4 hours after irradiation; group 6: after irradiated with 9 Gy, mice were injected with C57BL/6(H-2b) NK cells (1 x 10(6)) and 4 hours later with BM cells, group 7: after irradiation of 9 Gy, mice were injected with C57BL/6(H-2b) NK cells (1 x 10(6)) and 4 hours later with BM cells and spleen cells. The blood routine test, survival time, body weight, and histopathology in the recipients were observed and compared among these group. RESULTS: The survival time was (10.1 +/- 0.9), (9.8 +/- 0.9), (22.7 +/- 3.2) and (20.1 +/- 1.7) days in groups 1, 2, 3, and 5 respectively; was (30.1 +/- 16.0) days in group 4, out of which 2 mice survived for more than 30 days. The survival time was (39.1 +/- 18.1) and (49.3 +/- 17.2) days in groups 6 and 7 respectively, out of which 4 mice in group 6 and 7 mice in group 7 survived for more than 30 days. The survival time in group 6 was much longer than that in group 1, 2, 3 and 5 (P < 0.01). The survival time in group 7 was much longer than that in other groups (P < 0.05). The liver and spleen enlargement, organ destruction and infiltration with leukemic cells were observed in mice died from leukemia. The chimerism of Y chromosome appeared (80%-90%) in long-term survival mice in groups 6 and 7. CONCLUSION: Donor-derived NK cells have the antileukemia ability and reduce GVHD in haploidentical BMT in erythroleukemia mice (EL9611, H-2d). PMID- 19563022 TI - [Common variable immunodeficiency: report of two cases and literature review]. AB - OBJECTIVE: To promote the awareness of common variable immunodeficiency (CVID). METHODS: Report two cases of CVID and review related literature. RESULTS: The two CVID patients were manifested recurrent infections of respiratory and digestive tract, splenomegaly, significantly reduced serum immunoglobulin, since adolescence. They were treated with intravenous gammaglobulin, anti-infection and symptomatic therapies and the diseases were well controlled. Case 1 was diagnosed as with spleen peripheral T cell lymphoma after splenectomy. Case 2 developed nutritional megaloblastic anemia resulting from repeated digestive tract infections. CONCLUSION: CVID is a heterogeneous group of diseases with later onset, characterized by hypogammaglobulinemia and recurrent bacterial infections. Early diagnosis and regular treatment may improve the prognosis. PMID- 19563023 TI - [The role of PTEN-FAK signaling pathway in metastasis and invasive ability of leukemia cells]. AB - OBJECTIVE: To investigate the effect of the wild type phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a tumor-suppressor gene on the proliferation and apoptosis of human chronic myeloid leukemia (CML) cells line (K562) in vitro and explore the influence of PTEN-FAK signaling pathway on invasion and metastasis of leukemia cells. METHODS: The recombinant Ad-PTEN gene containing green fluorescent protein gene (Ad-PTEN-GFP) or the empty vector (Ad GFP) was transfected into K562 cells and fresh leukemia cells from CML patients in blast crisis. The growth of K562 cells was assayed by MTT assay; the apoptosis rate was assessed by flow cytometry (FCM). PTEN and FAK mRNA levels were detected by real-time fluorescent relative- quantification reverse transcriptional PCR (FQ PCR) and its protein levels by Western blot. The metastasis and invasive ability was examined by transwell chamber assay. RESULTS: The growth of K562 cells was suppressed markedly when Ad-PTEN-GFP was transfected into K562 cells at the 200 multiplicity of infection (MOI). The maximum growth inhibition rate was 35.2%. Transwell results showed the number of cells entered the lower chamber in Ad-GFP group was 9.1 fold more than that in Ad-PTEN-GFP group;The ability of metastasis and invasion of fresh leukemia cells was also suppressed after transfection with Ad-PTEN-GFP. FAK and p-FAK proteins were down-regulated by 0.72 and 0.16 fold lower after transfected with Ad-PTEN-GFP compared with Ad-GFP group. CONCLUSIONS: PTEN gene might inhibit the proliferation, metastasis and invasive ability of leukemia cells via down-regulating FAK expression. PMID- 19563024 TI - [In vitro cytotoxic effects of CML28 specific T cells on acute leukemia cells]. AB - OBJECTIVE: To explore the activation and proliferation of specific T cells induced by artificial antigen-presenting cells (aAPCs) simulated dendritic cells (DCs) and to observed the effect of these T cells on leukemic cell killing. METHODS: aAPCs were developed by coating a human leukocyte antigen-immunoglobulin fusion protein ( HLA-lg), which was connected each one of the four CML28 antigen epitopes (DLMSSTKGL, DLMSSTKGL, ALFCGVACA, VLTFALDSV), and CD28-specific antibody, to magnet-beads CML cell specific peptides (CML28) served as target peptides. Bone marrow (BM) or peripheral blood (PB) mononuclear cells (MNCs) were isolated from HLA-A2 healthy volunteers, and co-cultured with aAPCs. Specific T lymphocyte were detected by flow cytometry. The fresh acute leukemic cells were used as target cells. The specific T cells incubated with leukemic cells for 4 h at ratios of 5:1, 10:1, 20:1, 40:1, 80: 1, respectively. The effect of leukemic cells killing was detected by lactate dehydrogenase release test. RESULTS: The average ratio of CML-28 specific T lymphocyte in control group was (2.2 +/- 0.4)% and in experimental groups (DLMSSTKGL, DLMSSTKGL, ALFCGVACA, VLTFALDSV) were (13.5 +/- 1.6)%, (15.2 +/- 1.5)%, (14.7 +/- 1.8)% and (34.3 +/- 3.5)%, respectively, being significantly higher than that in control group (P < 0.01). Induction efficiencies of acute leukemic cells killing were significantly enhanced by increase of effector cells. The cytotoxic activity of specific T lymphocyte in one experimental group (VLTFALDSV) was much higher than that in other three experimental group (P < 0.05). CONCLUSION: This "prime and expand" regimen should be an alternative method for large scale amplification of rare tumor-specific CTLs and aAPCs might be a useful tool for leukemia immunotherapy. PMID- 19563025 TI - [To standardize the stratified therapy of postremission after induction therapy in adult acute myeloid leukemia patients]. PMID- 19563026 TI - [Re-evaluation of classification of myelodysplastic syndromes with low percentage bone marrow blasts]. AB - OBJECTIVE: To apply the WHO criteria and the minimal diagnostic criteria to the classification of myelodysplastic syndromes (MDS) with low percentage (< 0.050) bone marrow (BM) blasts. METHODS: Two hundred and ten MDS patients with less than 0.050 BM blasts diagnosed between 1988 and 2005 according to FAB criteria were retrospectively reclassified with WHO criteria (2001) and minimal diagnostic criteria. RESULTS: According to the WHO criteria, 5 patients were diagnosed as refractory anemia (RA), 7 as refractory anemia with ringed sideroblasts (RARS), 76 as refractory cytopenia with multilineage dysplasia (RCMD), 9 as RCMD-RS, 35 as MDS-unclassified (MDS-U), 3 as 5q - syndromes, and the rest 75 patients could not be classified suitably. Among the latter 75 patients 16 BM smears showed dysplasia in more than 2 cell lineage but only unilineage cytopenia in peripheral blood (PB). Nine of them were reclassified as RCMD after followed up for more than half a year. Forty-four BM smears showed erythroid dysplasia only, but bicytopenia or pancytopenia in PB. Twenty-seven of them were classified as RCMD after follow-up. Fifteen BM smears not showed dysplasia in any myeloid lineage were reclassified as MDS (5 patients), HS-MDS (5 patients) and idiopathic cytopenia of uncertain significance (ICUS) (5 patients) according to the MDS minimal diagnostic criteria. CONCLUSION: According to WHO criteria (2001), RA is the least diagnosis in MDS. The minimal diagnostic criteria for MDS classification of patients not fulfilled the standard criteria of MDS. PMID- 19563027 TI - [Lipoprotein lipase and serum thymidine kinase level in chronic lymphocytic leukemia and their correlations with other prognostic factors]. AB - OBJECTIVE: To investigate lipoprotein lipase (LPL) and serum thymidine kinase (TK) levels in chronic lymphocytic leukemia (CLL) and their correlations with other prognostic factors. METHODS: LPL expression level in peripheral blood samples of 58 CLL patients was detected by semi-quantitative reverse transcription PCR (RT-PCR). Serum TK1 level in 39 CLL patients was detected by enhanced chemiluminescence (ECL) and TK monoclonal antibody (Anti-TK mAb). IgVH mutation status was detected by multiplex PCR and sequencing of purified PCR products. The expression of ZAP-70 protein and CD38 were determined by flow cytometry . Panel probes and fluorescence in situ hybridization (FISH) were used to detect cytogenetic aberrations. RESULTS: The median LPL expression level in CLL was 0.26 (0 -6.29), while undetectable in normal controls. LPL expression level was significantly correlated with IgVH mutation status, Binet stages, CD38 and cytogenetic aberrations. Patients with unmutated IgVH genes had higher LPL than those with IgVH mutations (P = 0.010). Patients in Binet stage B and C had higher LPL than those in stage A (P = 0.011). LPL level was higher in patients with CD38 > or = 30% (P = 0.001). Higher LPL level was found in patients with unfavorable cytogenetic aberrations (deletion in 17p13 or 11q22) than those with favorable cytogenetics (deletion in 13q as the sole abnormality) (P = 0.002). LPL level was not significantly correlated with sex, age, and ZAP-70 protein (P > 0.05). The level of TK1 was higher in CLL patients than that in normal control (P < 0.05). Patients with higher level of absolute lymphocyte count (ALC), lactate dehydrogenase (LDH), unmutated IgVH genes and ZAP-70 had higher levels of TK1 than those with lower level of ALC, LDH, mutated IgVH genes and ZAP-70 (P = 0.018, P = 0.018, P = 0.030 and P = 0.038, respectively). TK1 level had no correlation with sex, age, Binet stages, CD38, and cytogenetic aberrations (P > 0.05). CONCLUSIONS: LPL expression and serum TK1 levels significantly correlate with other CLL prognostic factors and could be predictive markers for IgVH mutation status. LPL and serum TK1 might be applied to the assessment of prognosis in CLL patients. PMID- 19563029 TI - [Study on the clinical characteristics of adult biphenotypic acute leukaemia]. AB - OBJECTIVE: To analyze the clinical and biological characteristics and prognosis of adult biphenotypic acute leukaemia (BAL). METHODS: Immunophenotypes were analyzed using multicolor flow cytometry, karyotype analysis by short-term culture R-banding technique. The chemotherapy regimens were accordingly for acute lymphoblastic leukaemia (ALL), acute myeloid leukaemia (AML) or for both ALL and AML. Patients with Ph (+) or bcr-abl (+) were treated with Imatinib. RESULTS: (1) The incidence of BAL in acute leukaemias was 6.7%, with a male predominance and 52.3% of BAL patients had WBC > or = 30 x 10(9)/L and 16.9% WBC > or = 100 x 10(9)/L. (2) Percentages of coexpression of myeloid and B lymphoid antigens were 81.5%, of myeloid and T lymphoid antigens 10.8%, of myeloid, B- and T lymphoid antigens 4.6%, and of B and T lymphoid antigens 3.1%. (3) Normal and abnormal karyotypes accounted for 41.5% and 58.5%, respectively in 53 BAL patients with karyotype analysis. The rate of Ph (+) or bcr-abl (+) was 32.1%. (4) 31 (56.4%) of 65 patients achieved complete remission (CR), but CR rate was only 35.3% for Ph (+) or bcr-abl (+) cases. CONCLUSION: (1) High white blood cell count and coexpression of myeloid/B lymphoid antigens are common in BAL. (2) Abnormal karyotypes and Ph (+) or bcr-abl( +) often happen. (3) The treatment outcome of BAL is poor. PMID- 19563028 TI - [Arsenic trioxide inhibits cell growth in imatinib-resistant bcr-abl mutant cell lines in vitro]. AB - OBJECTIVE: To explore the effect of arsenic trioxide (As2O3) on the growth inhibition of imatinib (IM)-resistant bcr-abl mutant cell lines in vitro. METHODS: Cell growth of one IM-sensitive cell line, 32Dp210 and 15 IM-resistant cell lines including T315I and other 14 bcr-abl mutants were detected by MTT assay after treatment with IM and As2O3. The cell lines with 5 frequently observed mutants in CML patients were analyzed for apoptosis by flow cytometry with Annexin V and PI staining as well as the expression of bcr-abl fusion protein, phosphorylated CRKL protein and apoptosis-related proteins by Western blot. RESULTS: The fifty percent inhibition concentration (IC50) values of As2O3 for 15 IM-resistant cell lines were 2.6-5.3 fold lower than that for IM-sensitive cell line. For the 5 bcr-abl mutants frequently happened in CML patients, As2O3 significantly inhibited the expression of bcr-abl fusion protein and phosphorylated CRKL and induced apoptosis in a dose-dependent manner as compared with that for 32Dp210. Coincidently, the cell apoptosis was induced through caspase-3, 8 and 9 pathways. CONCLUSION: As2O3 remarkably inhibits cell growth and induces apoptosis of IM-resistant bcr-abl mutant cell lines in vitro, suggesting that it might be a potential therapeutic agent for IM-resistant CML patients. PMID- 19563030 TI - [Efficacy of IA regimen followed by FLAG regimen in the treatment of acute myeloid leukaemia]. AB - OBJECTIVE: To investigate the efficacy and toxicity of standard-dose IA regimen (idarubicin and cytarabine) as induction therapy followed by FLAG regimen in patients with acute myeloid leukemia (AML), and its influence on peripheral stem cell mobilization. METHODS: A total of 23 previously untreated de novo AML patients were enrolled. Thirteen patients were male, and 10 female, with ages ranging from 14 to 54 (median: 41) years. Cytogenetic analysis was performed for all patients. The IA regimen contained idarubicin (12 mg x m(-2) x d(-1), days 1 to 3) and cytarabine (100 mg x m(-2) x d(-1), days 1 to 7), and the FLAG regimen contained'fludarabine (50 mg/d, days 1 to 5), cytarabine (2 g x m(-2) x d(-1), days 1 to 5) and granulocyte colony-stimulating factor (G-CSF, 300 microg/d, days 0 to 5). RESULTS: After one course of induction therapy, the CR rate was 91.3%. The CR rate for patients with favourable and intermediate prognostic karyotypes was 100% and 91.3%, respectively. Nineteen patients in CR were consolidated with FLAG regimen, of which 6/9 (66.7%) patients were able to mobilize a sufficient number of CD34+ cells and successfully performed autologous stem cell transplantation. Four patients relapsed. The median survival duration was 19.5 months and median disease-free survival was 14 months. Myelosuppression and infections due to neutropenia were the most frequent adverse effects, severe nonhematologic toxicity and the early death were not observed in all patients. CONCLUSION: IA followed by FLAG regimen is effective and well tolerable in AML patients especially in those with favourable and intermediate prognostic karyotypes, and 1 to 2 courses of this therapy shows no influence on peripheral stem cell mobilization and subsequent autologous stem cell transplantation. PMID- 19563031 TI - [ALL-XH-99 protocol in the treatment of childhood T-cell acute lymphoblastic leukemia]. AB - OBJECTIVE: To analyze the incidence, clinical characteristics and prognosis of childhood T-cell acute lymphoblastic leukemia (T-ALL). METHODS: From January 1999 to April 2005, 305 patients with newly diagnosed ALL were enrolled in protocol ALL-XH-99. The clinical characteristics of these children were analysed. RESULTS: Of 305 childhood ALL patients, 43 were T-ALL. There were 34 males among the 43 T ALLs. The mean age at diagnosis was 7.8 (2.2 to 16.4) years, 29 (67.4%) cases of them were older than 10 years, and 27 (62.8%) cases had initial WBC count more than 50 x 10(9)/L. In comparison with that of B cell ALL (B-ALL), the percentages of age older than 10 years, initial WBC count more than 50 x 10(9)/ L, prednisone poor response (PPR), and failed to achieve remission at day 19 of induction chemotherapy in the T-ALLs were all higher. No statistic difference was found in sex between them. The eight-year event-free survival (EFS), relapse-free survival (RFS) and overall survival (OS) were (40.2 +/- 10.1)%, (51.4 +/- 11.6)% and (49.8 +/- 9.9)% for T-ALL, and (72.1-3.0)%, (83.2 +/- 2.7)%, and (76.6 +/- 2.9)% for B ALL, respectively, being differed significantly between the two types of ALL (P < 0.01). CONCLUSION: There were statistic differences between T-cell and B-cell childhood ALLs in age, initial WBC count, early response to therapy, and eight year EFS and RFS. Childhood T-ALL was associated with a worse prognosis than other sub-types of childhood ALL. PMID- 19563032 TI - [Experimental study of effect of As2S3 nanoparticles on human MDS cell line (MUTZ 1)]. AB - OBJECTIVE: To in vitro study the inhibition effect and possible mechanism of As2S3 nanoparticles (As2S3 nano) on human MDS cell line MUTZ-1 and to compare with that of traditional As2S3. METHOD: MUTZ-1 cells were treated with As2S3 nano and traditional regular-sized particles (TRSP) at different concentrations. The cell growth inhibition rate was determined by MTT assay, cell apoptosis by morphology and flow cytometry (FCM), cell cycle by FCM and the activity of caspase-3 by chemiluminescence assay. RESULTS: Treatment of As2S3 nano and TRSP at concentrations of 2, 4, 8 and 16 micromol/L for 48 h could lead to a significant dose-dependent decrease of MUTZ-1 cells and induce apoptosis. The percentages of inhibition were 48.9%, 75.9%, 89.4% and 96.5% in As2S3 nano vs 14.5%, 25.4%, 34.7% and 51.5% in TRSP and apoptosis rates were (12.9 +/- 1.9)%, (19.2 +/- 2.2)%, (30.1 +/- 2.5)% and (45.9 +/- 2.3)% in As2S3 nano vs (5.3 +/- 1.8%)%, (11.1 +/- 2.6)%, (19.3 +/- 2.3)% and (25.5 +/- 2.5)% in TRSP respectively. There was statistically significant difference in these two groups (P < 0.01). The proportion of cell in G2/M phase and the activity of caspase-3 of MUTZ-1 cells treated with A2S32 nano were significantly higher than those treated with control group and As2S3 TRSP groups (P < 0.01). CONCLUSIONS: As2S3 nanoparticles and TRSP can inhibit the proliferation of MUTZ-1 cells and induce apoptosis, which maybe through activating caspase-3 pathways and increasing the proportion of G2/M phase. As2S3 nanoparticles can produce a much better antitumor effect than As2S3 TRSP do. PMID- 19563033 TI - [Reversal of multidrug resistance of the drug resistant human multiple myeloma cell line MOLP-2/R by curcumin and its relation with FA/BRCA pathway]. AB - OBJECTIVE: To investigate the reverse effect of mutidrug resistance of curcumin combined with melphalan on the mutidrug-resistant human multiple myeloma cell line MOLP-2/R and the relation with FA/BRCA pathway. METHODS: The inhibitory effects of the drugs on the growth of MOLP-2/R cells were determined by MTT assay. Cell cycle analysis, intracellular drug concentration and apoptosis were assayed by flow cytometry. The expression of FANCD2 monoubiquitination was determined by Western blot analysis. RESULTS: Co-administration of curcumin and melphalan had an synergistic inhibitory effects on the proliferation, IC50 of melphalan with 10 micromol/L curcumin reduced from 45.5 micromol/L to 19 micromol/L in MOLP-2/R cells. The apoptosis percentage of MOLP-2/R cells was significantly increased from (23.3 +/- 0.6)% to (52.6% +/- 0.8)% by the treatment of melphalan 20 micromol/L plus curcumin 10 micromol/L with the increased percentage of cells in the G2/M phase (from 9.1% to 18.5%) and enhanced intracellular drug concentration of MOLP-2/ R cells (from 15.2 +/- 0.3 to 21.4 +/ 0.8 ). The effects were accompanied with inhibition of FA/BRCA pathway by down regulation of FANCD2 protein monoubiquitination. CONCLUSION: Curcumin combined with melphalan results in synergistic effects and reverses multiple drug resistance of MOLP-2/R cells effectively. The inhibition of FA/BRCA pathway may be the mechanism. PMID- 19563035 TI - [The status of anti-caries vaccine research and consideration]. PMID- 19563034 TI - [Effect of low-dose 2-methoxyestradiol on the expressions of two isoforms of PRDM1 gene in myeloma]. AB - OBJECTIVE: To investigate the effects of low-dose 2-methoxyestradiol (2ME2) on the expression of two isoforms of PRDM1 gene (PRDM1alpha with and PRDM1beta without PR-domain) during the myeloma cell differentiation. METHODS: In myeloma cell line NCI-H929, RPMI8226, KM3 and LP-1, PRDMla and PRDM1beta transcripts were detected by real-time quantitative PCR after treatment with low-dose of 2ME2 (0.5 micromol/L) for 0 h, 48 h and 72 h. RESULTS: Both PRDM1alpha and PRDMIbeta3 were time-dependently upregulated, and the PRDMlalpha/beta ratio was elevated from 1.461 +/- 0.033 to 2.663 +/- 0.381 (P < 0.01), 1.929 +/- 0.334 to 2.727 +/- 0.362 (P < 0.05), 1.471 +/- 0.012 to 4.367 +/- 0.243 (P < 0.01) and 1.660 +/- 0.042 to 3.059 +/- 0.167 (P < 0.01) in NCI-H929, RPMI8226, KM3 and LP-1 cells respectively. CONCLUSION: Low dose of 2ME2 can induce differentiation of myeloma cells as well as up-regulate the expression of PRDMlalpha and PRDM1beta in these cells. Elevation of PRDM1alpha/beta ratio was in a time-dependent manner and positively associated with cell differentiation and in accordance with Ying-Yang mechanism of PR-domain-containing gene family. PMID- 19563036 TI - [Tendency of caries prevalence in China and the strategies of handling]. PMID- 19563037 TI - [Correlation between Streptococcus mutans level in saliva and caries status in children]. AB - OBJECTIVE: To investigate the distribution pattern of salivary Streptococcus mutans (Sm) level in children and its association with caries status. METHODS: Three hundred and sixty-five 11- 12-year-old children participated in this study. Scores of decayed, missing or filled teeth (DMFT) for the permanent dentition were recorded. Salivary Sm concentrations were determined by species-specific monoclonal antibodies. RESULTS: Caries-affected children harbored more Sm in saliva [5.53 (1.50, 18.00) x 10(7)/L] than their caries-free counterparts [3.42 (1.60, 8.10) x 10(7)/L] (P = 0.002). Salivary Sm concentration tended to rise with the increase of DMFT score. Spearman's correlation coefficient for Sm concentration was 0.136 (P = 0.010). When salivary Sm concentration reached 8.64 x 10(7)/L, children's caries experience doubled. CONCLUSIONS: Salivary Sm level has a skewed distribution among the children studied and associates positively with caries presence. Determination of salivary Sm levels could be used to predict children's caries status. PMID- 19563038 TI - [Clinical evaluation of periapical endodontic surgery for endodontic failure]. AB - OBJECTIVE: To evaluate the clinical outcome of periapical endodontic surgery for teeth that can't be treated by nonsurgical endodontic methods. METHODS: Sixty-two affected teeth were chosen for surgical endodontic treatment, of which 31 teeth underwent periapical curettage and the others were treated by root-end resection, retrograde preparation and filling. A radiography was taken immediately after surgery and was compared with those taken at 12 and 24 months. The results of two groups were analyzed using the chi2 test. RESULTS: The success rate for retrograde filling was higher (85% after 12 months, 88% after 24 months) compared with that of periapical curettage (52% after 12 months, 45% after 24 months). The difference in success rate between the two groups was statistically significant. CONCLUSIONS: Ultrasonic root-end preparation and retrograde filling is a good choice of treatment when the teeth can't be treated appropriately by nonsurgical treatment. PMID- 19563039 TI - [Relationship of concentration of lactoferrin and lysozyme in saliva and dental caries in primary dentition]. AB - OBJECTIVE: To explore the relationship between the concentrations of lactoferrin and lysozyme in saliva and dental caries in primary dentition among Chinese children. METHODS: Forty children with high dmft score (dmft > or = 5) and 40 caries-free children (dmft = 0) were sampled and assigned into two groups. Total salivary proteins was measured by means of bicinchoninic acid. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was performed to analyze the images of target straps. Lactoferrin and lysozyme were detected using Western blotting method. RESULTS: The total salivary protein in high dmft group [(852.02 +/- 206.14) mg/L] was lower than that of caries-free group [(1032.44 +/- 221.99) mg/L, P < 0.001]. The ratio of 77,000 protein in high dmft group [(12.50 +/- 7.73) IA/microg] was significantly higher than that of the caries-free children [(8.71 +/- 4.28) IA/microg, P = 0.009], while there was no significant difference for 14,500 protein between them (P = 0.137). The ratio of lactoferrin was higher in high dmft group [(229.04 +/- 197.14) IA/microg] than that in caries-free children [(144.07 +/- 99.91) IA/microg, P = 0.018], while no significant difference for lysozyme between the two groups (P = 0.091). CONCLUSIONS: Saliva protein is closely related to caries in primary dentition. Lactoferrin may be one of the important components. PMID- 19563041 TI - [Bacterial quorum sensing system and its signal transmission]. PMID- 19563040 TI - [A proteomic analysis of human dental pulp cells undergoing odontoblast differentiation]. AB - OBJECTIVE: To analyze the difference in protein profiles between human dental pulp cells (DPC) and odontogenic differentiated DPC by using proteomic approach. METHODS: Human DPC were induced to odontoblast differentiation and total proteins in the cell lysates before and after induction were prepared. Proteins spots were isolated by two-dimensional gel electrophoresis. DeCyder V6.0 software was applied to gel image analysis. Differential protein spots were identified by peptide mass fingerprinting technique. RESULTS: Forty-six protein spots were determined to be differentially expressed with twenty identified protein spots. Expression changes of the identified proteins revealed the involvement of various regulation mechanisms in odontoblast differentiation, such as cell cycle, cellular energy regulation and signal transduction. CONCLUSIONS: The proteomic approach is a high throughput method to screen the candidate proteins involved in odontoblast differentiation of DPC. PMID- 19563042 TI - [Reliability of superficial temporal artery and vein as the recipient vessels in free flap transfers in head and neck region]. AB - OBJECTIVE: To examine the reliability of superficial temporal artery and vein as the recipient vessels in free flap transfers in head and neck region. METHODS: From May 2001 to June 2008, 25 cases of head and neck reconstructions were performed by using the vessels in the temporal region as recipient vessels. RESULTS: Among the 25 recipient veins, 23 were superficial temporal veins, and 2 were deep temporal veins. The recipient arteries of all 25 free flaps were superficial temporal arteries. All the free flaps survived completely, without postoperative vessel thrombosis. CONCLUSIONS: The superficial temporal artery and vein can be used as reliable recipient vessels in free flap transfers in head and neck region. PMID- 19563043 TI - [Analysis of iatrogenic factors related to delay in diagnosis of oral squamous cell carcinoma]. AB - OBJECTIVE: To investigate the clinical iatrogenic factors related to delay in diagnosis of oral squamous cell carcinoma. METHODS: A total of 102 consecutive patients with squamous cell carcinoma of the oral cavity, treated in Department of Oral and Maxillofacial Surgery, Peking University Hospital of Stomatology were included in the study. For iatrogenic (professional) delay, stepwise multiple logistic regression and monofactor analysis, chi-square, test with 95% confidence interval (CI), were applied to analyze the correlation between the professional delay in diagnosis and the possible related clinical factors. RESULTS: Whether adopting biopsy at the first clinical management, length of time from the first clinical visit to adopting biopsy, number of clinical departments visited and whether taking any treatment before definitive diagnosis had significant influence on the delay. CONCLUSIONS: Taking biopsy at the first clinical visit, shortening the time from the first visit to biopsy and less undirected treatment can reduce the iatrogenic delay. PMID- 19563044 TI - [The study of cyclic fatigue and lifetime for all-ceramic crown after cementation]. AB - OBJECTIVE: To study mechanical and cyclic fatigue behavior of IPS Empress2 under cyclic loading, and to establish guidelines for the use and design of all-ceramic crowns. METHODS: A 3-D finit element method model of tooth and crown were established. The strength and lifetime of all-ceramic crowns under cyclic loading in centric occlusion were investigated using computational techniques of the Abaqus and MSC Fatigue software. RESULTS: Most of the fatigue and fracture of all ceramic crown occurred within the veneering material at cervical marginal of the crown. The number of loading cycles before failure occurred varied within specified limits 2,506,109-6,950,243. The lifetime of the crown decreased significantly as loading increased and decreased gradually as loading time increased as well. CONCLUSIONS: The mechanical and fatigue behavior of ceramic materials and restorations need to be improved before clinical use in order to guarantee clinical long-term success of all-ceramic crown. properly in order to increase the longevity of all-ceramic crowns. PMID- 19563045 TI - [Investigation of the color of anterior teeth, vermilion and attachment gingiva of 151 Uygur youth]. AB - OBJECTIVE: To analyze the characteristics of color of Uygur's oral soft and hard tissue. METHOD: One hundred and fifty-one Uygur medical students (18 to 22 years) were included in this study. The Lab*, Cab*, Hab0, values of subjects' anterior teeth, upper and lower vermilion, attachment gingiva were measured. RESULTS: The mean and distribution of Lab*, Cab*, Hab0, values of Uygur youth' anterior teeth were obtained. The Lab* values of attachment gingiva, upper and lower vermilion in male were 50.50 +/- 3.59, 41.92 +/- 4.10, 45.02 +/- 3.06, and Cab* were 29.94 +/- 3.21, 27.00 +/- 2.47, 30.35 +/- 2.67, and Hab0 were 30.00 +/- 3.27, 31.71 +/- 3.69, 27.73 +/- 3.27, respectively. The Lab* values of attachment gingiva, upper and lower vermilion in female were 54.28 +/- 5.06, 41.73 +/- 3.70, 47.27 +/- 2.97, and Cab* were 27.98 +/- 3.93, 27.23 +/- 2.42, 31.22 +/- 2.59, and Hab0 were 33.39 +/- 3.67, 32.28 +/- 3.68, 27.57 +/- 2.65. CONCLUSIONS: There was a specific color space distribution pattern in oral soft and hard tissue in Uygur youth. There was a trend of decreasing lightness, increasing saturation, and red hue shifting from the center to both distal directions. There was statistical significant gender difference in color of tooth and attachment gingiva. What's more, there was extremely significant difference between upper and lower vermilion. PMID- 19563046 TI - [Comparative ultrastructural study of parotid gland, lacrimal gland and pituitary gland between miniature pig and mouse]. AB - OBJECTIVE: To study the ultrastructure of parotid glands, lacrimal glands and pituitary glands between miniature pig and mouse. METHODS: Five adult miniature pigs and 5 mice were studied. Ultrastructure of their parotid glands, lacrimal glands, and pituitary glands was observed. RESULTS: The secretary granules in acinar cell of miniature pig parotid glands showed higher density and more aequalis than those of mice. The cell apparatus in acinar cell of mouse parotid glands were more plentiful than those of miniature pigs. The secretary granules on blood vessel wall were richer in parotid gland of miniature pigs compared with mouse parotid gland. Lacrimal gland had the similar ultrastructure to parotid gland in these two animals. Many blood vessel antrum were found in pituitary glands of these two animals. CONCLUSIONS: Compared with mouse parotid glands, there are more secretary granules in acinar cells and vascular endothelial cells in miniature pig parotid glands, which might enter blood stream and have function of endocrine secretion. PMID- 19563047 TI - [Comparison of shear bond strength of different resin cements to ceramic and dentin]. AB - OBJECTIVE: To evaluate the effect of different resin cements and dentin surface treatment methods on shear bond strength between glass ceramic and human dentin. METHODS: Freshly extracted non-carious human premolars were collected to prepare flat mesial and distal dentin surface with diamond bur. Besides, ceramic discs (3 mm in diameter, 3 mm in height) were fabricated with IPS e. max Press ingots. All samples were randomly divided into five groups, sixteen specimens each. Ceramic discs were cemented on dentin surface using one of the five resin cements [Group A (Variolink II), Group B (Multilink Automix), Group C (Multilink Sprint), Group D (Rely X Unicem) and Group E (BisCem)] respectively. Half of the specimens were stored in water 37 degrees C for 24 h and the other half were additionally thermocycled for 5000 times (5 degrees C/55 degrees C) before shear bond strength test. Dentin bonding surface and transverse plane were examined with a scanning electronic microscope. RESULTS: Group A showed the highest bond strength before thermocycling [(22.3 +/- 3.9) MPa], and the bond strength of Group B [(18.1 +/- 3.5) MPa] was lower than that of Group A (P < 0.05). Group E [(11.7 +/- 4.2) MPa], Group C [(11.3 +/- 3.6) MPa] and Group D [(14.1 +/- 2.3) MPa] had the lowest bond strength. After thermocycling, Group A still showed the highest bond strength [(17.8 +/- 2.3) MPa]. The bond strength of Group B [(14.4 +/- 3.5) MPa] and Group D [(13.2 +/- 2.5) MPa] were lower than that of Group A. Group E [(8.9 +/- 3.2) MPa] and Group C [(7.0 +/- 2.4) MPa] showed the lowest bond strength. The bond strength of all groups decreased notably after thermocycling except Group E and Group D. CONCLUSIONS: The resin cement based on etch-rinsing bonding system has higher bond strength to dentin than those based on self-etch bonding system and self-adhesive resin cements. PMID- 19563049 TI - [Rights maintenance and self-discipline of practicing dentist: part XVII. Responsibility for quoting proof in lawsuits for medical care dissension]. PMID- 19563048 TI - [The identification of electronic surveying lines on 3-D digital models of dentition defects]. AB - OBJECTIVES: To develop a mathematical algorithm and a software package for the process of electronically surveying a scanned point cloud cast. To provide a principal premise to the subsequent computer aided design and computer aided manufacture (CAD/CAM) of removable partial denture framework, and to provide a method to improve quality control in the dental laboratory. METHODS: Point cloud data of a partially edentulous cast, a mandibular Kennedy Class II Modification 2 arch, was captured using an optical scanning system with projective grating and high-resolution digital camera. Using commercial CAD/CAM software system (Geomagic Studio 6), this point cloud data was processed and the 3-D digital model of partially edentulous cast was reconstructed. From a suggested surveying angle the contour points of height were identified, and then the digital surveying lines were traced using Projection and Contour Extraction software package. The depth of undercut was measured and defined to determine the clasp termination of retainer. RESULTS: Electronic surveying line of 3-D digital dentition defect model was achieved. Digital surveying line defined the cast into undercut and non-undercut areas. Different virtualized paths of insertion could be automatically suggested when the cast was surveyed and analyzed from different angles. The depth of undercut was automatically measured and the retentive clasp termination was determined. CONCLUSIONS: The mathematical algorithm and the software package in this study can be used to survey and analyze 3-D digital models of dentition defects, and to identify an electronic surveying line. PMID- 19563050 TI - [Orthodontic treatment for adults: part I. Special treatment considerations and general process for adult orthodontics]. PMID- 19563051 TI - [To study and practice scientific outlook on development, accelerate sustainable development for all disciplines]. PMID- 19563052 TI - [Vaginal microflora and vagini]. PMID- 19563053 TI - [Microecological investigation of vaginal microflora in women with varying degree gynecologic symptoms in clinics]. AB - OBJECTIVE: To investigate the microecological status of vaginal microflora in the women with different vaginal symptoms. METHODS: From March 2006 to October 2007, 6982 cases with varying degree vaginal symptoms including pruritus, increaseed leucorrhea, the leucorrhea having unusual smell, in the gynecology outpatient department were studied. The vagina secretions were examined in terms of the pH value, the hydrogen peroxide test, and Gram dyeing inspection of vaginal bacteria and microecology appraisal for colony's density, the multiplicity, the superiority fungus, and the inflammatory response. RESULTS: Among 6982 patients, normal vaginal microecology was identified in 750 (10.74%, 750/6982); abnormal microecology was found in 6232 (89.26%, 6232/6982); bacterial vaginosis (BV) was detected in 729 (10.44%, 729/6982); vulvovaginal candidiasis (VVC) was in 1527 (21.87%, 1527/6982). Ninety five patients (1.36%, 95/6982) were with both BV and VVC. Abnormal bacteria colonies were found in 1229 (17.60%, 1229/6982), and others were found in 2652 (37.98%, 2652/6982). CONCLUSIONS: The vaginal microecology in the women with different vaginal symptoms can be either normal or abnormal. Microecology clinical evaluation system can assess the vaginal microecosystem. PMID- 19563054 TI - [Investigation of the situation of vaginal microflora in healthy women population]. AB - OBJECTIVE: To assess the distribution of vaginal microflora in healthy women and investigate their cognition of lower reproductive tract infection-related knowledge as well as personal hygiene habits and character of behavior in seeking medical treatment. METHODS: Total of 1660 healthy women who had physical check ups at the Medical Center of Renji Hospital were selected and received gynecologic examination, as well as routine examination, pH examination and bacterial culture of the vaginal discharge. In addition, 860 of them were randomly selected for questionnaire survey, in which reproductive tract infection -related knowledge and personal hygiene habits as well as behavior in seeking medical treatment were involved. RESULTS: Among the 1660 censused women, vaginal average pH was 4. 16 +/- 0.21. The positive rate of Candida in vaginal discharge routine examination was 3.86% (64/1660), which was lower than that in aerobes culture 7.71% (128/1660). Candida albicans was the most populous species 78.9% (101/128), followed by 7.8% (10/128) and 7.0% (9/128) for the Candida glabrata and Candida krusei respectively. The most populous species of vaginal microflora were hemolytic streptococcus A (63.80%, 1059/1660), Staphylococcus epidermidis (14.28%, 237/1660), and Enterococcus faecalis (D) group (11.44%, 190/1660). The questionnaire survey showed that 88.4% (760/860) of 860 women took active treatment when feeling unwell, 92.1% (792/860) of them had good hygiene practices, and only 21.2% (182/860) had the habit of vaginal douching. In addition, 50.0% (430/860) of them had the desire to obtain reproductive health knowledge through out-patient consultation. CONCLUSIONS: Vulvovaginal Candida disease ranks the first in all types of vaginitis, among which, Candida albicans is the most populous species followed by the Candida glabrata and Candida krusei. Hemolytic streptococcus A and Staphylococcus epidermidis are the most common species of vaginal microflora in healthy women. The censused women have high awareness of reproductive health care and pay much attention to common gynecologic diseases such as lower reproductive tract infection. Nevertheless, we should strengthen the public education of reproductive health-related knowledge. PMID- 19563055 TI - [Study of local immunity of lower genital tract infections]. AB - OBJECTIVE: To investigate the profile of local immunity of vagina and the immune defense mechanisms against lower genital tract infections. METHODS: Vaginal lavage was collected from healthy women and patients of vulvovaginal candidiasis, bacterial vaginosis, Trichomonol vaginitis, human papilloma virus infection (VVC), and chlamydia trachomatis infection. Each group included 60 cases. The level of interleukin (IL)2, 4, 5, 13, 8 and human defensin 5 (HD5) were detected by enzyme linked immunosorbent assay(ELISA). RESULTS: (1) Cytokine of helper T cell 1(Th1): the level of IL-2 between healthy women and VVC/ bacterial vaginosis (BV)/ trichomonol vaginitis (TV)/ chlamydia trachomatis (CT) patients had no significant difference. The IL-2 level(96 +/- 33) x 10(-3) pg/L of human papilloma virus (HPV) infection patients was significantly higher than that of healthy women (P < 0.05). (2) Cytokine of helper T cell 2 (Th2): the level of IL 4 between healthy women and VVC/CT patients had no significant difference. The level of IL-5 between healthy women and BV patients had no significant difference. The IL-13 level (42 +/- 15) x 10(-3) pg/L of TV patients was significantly higher than that of healthy women (30 +/- 29) x 10(-3) pg/L (P < 0.05). The IL-4 level (103 +/- 28) x 10(-3) pg/L of HPV infection patients was significantly higher than that of healthy women (36 +/- 22) x 10(-3) pg/L (P < 0.05). (3) IL-8: the IL-8 level (5.8 +/- 2.7) pg/L of TV infection patients was significantly higher than that of healthy women (2.6 +/- 2.4) pg/L (P < 0.05). The level of IL-8 between healthy women and BV patients had no significant difference. (4) HD5: the HD5 level of TV, BV, VVC, HPV and CT infection patients were significantly higher than that of healthy women (P < 0.05). CONCLUSIONS: (1) HD5 plays an important role in the defence of vaginal epithelial cell. (2) Th2 may be more important than Thl in lower genital tract infections. (3) IL-8 plays an important role in extrinsic source infections. PMID- 19563056 TI - [Status and future of fetal-maternal medicine]. PMID- 19563057 TI - [Expression of aquaporin 8 in human fetal membrane and placenta of idiopathic polyhydramnios]. AB - OBJECTIVE: To determine the expression of Aquaporin 8(AQP8) in the fetal membrane and placenta of idiopathic polyhydramnios. METHODS: The amnion, chorion and placenta were collected from 12 term pregnancies with idiopathic polyhydramnios( polyhydramnios group) and 12 term pregnancies who were normal (control group). The expression of AQP8 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of AQP8 protein was detected by immunohistochemistry. RESULTS: The expression of AQP8 mRNA in amnion, chorion and placenta of polyhydramnios group was (0.78 +/- 0.13), (0.58 +/- 0.10), and (0.86 +/- 0.15) respectively, and that of control group was (0. 39 0.07), (0.45 +/- 0.09), and (0.34 +/- 0.09) respectively. The expression of AQP8 protein in amnion, chorion and placenta of polyhydramnios group was (0.195 +/- 0.024), (0.170 +/- 0.028), and (0.193 +/- 0.024) respectively, and that of control group was (0.151 +/- 0.018), (0.156 +/- 0.024), and (0.152 +/- 0.023) respectively. In all 3 types of tissues the expression of AQP8 mRNA of polyhydramnios group was higher than that of control group (P < 0.05). In amnion and placenta the expression of AQP8 protein of polyhydramnios group was also increased compared to that of control group (P < 0.05), but in chorion the difference in AQP8 protein expression between the two groups was not significant (P > 0.05). CONCLUSION: The expression of AQP8 mRNA and protein is significantly increased in the amnion and placenta of polyhydramnios, suggesting that AQP8 may play an important role in the regulation of amniotic fluid volume. PMID- 19563058 TI - [Relationship of fetal total bile acid and the change of fetal pancreas endocrine secretion and its impact on fetal growth and development in intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To investigate the relationship of fetal total bile acid (TBA) concentration with the change of fetal pancreas endocrine secretion and its impact on fetal growth and development in intrahepatic cholestasis of pregnancy (ICP). METHODS: The concentrations of TBA, insulin, glucagon and glucose in the cord blood were measured in 30 fetuses with maternal ICP (case group) and 30 fetuses of normogravidas (control group) after elective cesarean section during the same period in the Department of Obstetrics of Xiangya Second Hospital of Central South University from March 2007 to February 2008. The cord blood TBA concentration was investigated by enzyme method and the concentrations of insulin and glucagon were investigated by radioimmunoassay. The glucose was measured by oxidase-superoxide method. The neonatal weight, length and the ponderal index (PI) were measured after parturition. RESULTS: (1) The cord blood insulin concentration (9.0 +/- 3.3) mU/L and the ratio of insulin over glucagon 0.048 +/- 0.028 in the case group was significantly lower than that of controls (10.1 +/- 3.7) mU/L, 0.050 +/- 0.020 (P < 0.05). The concentrations of TBA (10.3 +/- 3.8) miromol/L and glucagon (235 +/- 57) ng/L in case group were obviously higher than that in controls (4.1 +/- 1.3) micromol/L, (205 +/-34) ng/L (P < 0.05). But no difference was shown in the glucose concentration in cord blood between the case and control groups [(3.4 +/- 1.1) mmol/L vs. (3.6 +/- 1.2) mmol/L, P > 0.05]. (2) The neonatal weight and length in case group were significantly lower than that of control [(3163 +/- 478) g vs. (3498 +/- 393) g, (46.5 +/- 2.3) cm vs. (49.3 +/ 1.9) cm, P < 0.01]; while the Ponderal index in case group was significantly higher than that of control group (3.13 +/- 0.23 vs. 2.92 +/- 0.29, P < 0.01). (3) The cord blood TBA concentration respectively showed a linear relationship with the cord blood insulin concentration, the cord blood glucagon concentration and the ratio of insulin over glucagon in the case group. With the increase in cord blood TBA concentration, the cord insulin concentration and the ratio of insulin over glucagon decreased; meanwhile the cord blood glucagon concentration rose (P < 0.01). The cord blood insulin concentration and the ratio of insulin over glucagon in case group were respectively positively correlated with the neonatal weight and length, and were negatively correlated with the PI (P < 0.01); while the cord glucagon concentration was respectively negatively correlated with the neonatal weight and length, and positively correlated with the PI (P < 0.01). CONCLUSIONS: In ICP fetus pancreas, there are hypoinsulinism, glucagon oversecretion, and decrease of the ratio of insulin over glucagon, which is closely correlated with fetal TBA concentration. The endocrine function of fetal pancreas affects the fetal growth and development. PMID- 19563059 TI - [Ultrasound-guided transvaginal ovarian interstitial laser treatment in patients with polycystic ovary syndrome: a laser dose-finding study]. AB - OBJECTIVE: To evaluate the clinical and endocrine effectiveness of different laser doses for ultrasound-guided transvaginal ovarian interstitial laser treatment in patients with polycystic ovary syndrome (PCOS). METHODS: Between January 2005 and July 2007, 56 women with clomifene citrate-resistant PCOS selected from the patients who were referred to Shenzhen Maternity and Child Healthcare Hospital with a request for fertility underwent ultrasound-guided transvaginal ovarian interstitial laser treatment. All subjects were randomly divided into four groups of A, B, C and D. In group A, one coagulation point per ovary was done and group B, two points; group C, three points; group D, four to five points. The size of each point was about 10 mm in diameter (the electrical laser was projected persistently for 1-3 min with a power of 3 -5 W). The serum sexual hormone level, ovulation rate and pregnancy rate within six postoperative months were compared among the four groups. RESULTS: (1) The spontaneous ovulation rates of groups A (0) and B (21%) within six postoperative months were significantly lower than groups C (71% ,P <0. 05) and D (79%, P < 0.01). The accumulative pregnancy rates of group C(43%) and D(36%) for six postoperative months were significantly higher than group A (0; P < 0.01, P < 0.05). Although they were also higher than that of group B, no statistical significance was found (P > 0.05). (2) No statistically significant differences were found among four groups when various preoperative hormone values were compared (P > 0. 05). The mean serum luteinizing hormone (LH), testosterone level and LH/ follicle stimulating hormone (FSH) ratio was significantly lower postoperatively in groups C [(6.3 +/- 2.6) U/L, (2.2 +/- 0.7) nmol/L, 1.1 +/- 0.3] and D [(5.8 +/- 2.5) U/L, (2.1 +/- 0.4) nmol/L, 1.0 +/- 0.4] than in groupsA [(11.9 +/- 3.1) U/L, (3.9 +/- 1.6) nmol/L, 2.1 +/- 0.5] and B [(10.4 +/- 3.9) U/L, (3.3 +/- 1.1) nmol/L, 2.0 +/- 0.6], respectively (P < 0.05). The mean LH, testosterone level and LH/FSH ratio reduced more obviously in groups C (42%, 39% and 42%) and D (53%, 40% and 58%) than in groups A (4%, 9% and 16%) and B (11%, 6% and 5%; P < 0.05). All above-mentioned parameters between groups C and D had no statistical significant difference (P > 0.05). CONCLUSIONS: One and two intraovarian laser coagulation points per ovary are associated with poor results. Three points per ovary seem to represent the plateau of effective dose for the ovarian interstitial laser treatment. Increasing the dose above it does not improve the outcome. PMID- 19563060 TI - [Bacterial composition and resistance from urinary tract infections in females]. AB - OBJECTIVE: To determine the bacterial composition and antibacterial resistance in female urinary tract infections (UTI) in China. METHODS: The disc diffusion test, minimum inhibition concentration (MIC), test or E-test was used to detect the antimicrobial susceptibility in 86 tertiary hospitals in China from June 1, 2006 to May 31, 2007. WHONET 5.4 was applied for data analysis. RESULTS: A total of 6071 pathogenic germs were collected from urine cultures of UTI female patients in the surveillance period, which included 5958 bacterial isolates. Escherichia coli 3529 strains (59.23%), Enterococcus spp 938 strains (15.74%) and Klebsiella pneumoniae 394 strains (6. 61%) were the most common pathogens. Escherichia coli and Klebsiella pneumoniae had 35.7% and 34.0% extended spectrum beta-lactamases (ESBL) positive rates, and their resistant rates to quinolones were about 70% and 40%, respectively. Enterococcus faecium was much more resistant to almost all the antibiotics than Enterococcus faecalis. 0.9% Escherichia faecalis and 3. 8% Escherichia faecium were resistant to vancomycin, whereas 2.7% and 4.8% were resistant to teicoplanin. Staphylococcus epdermidis was more common in UTI than Staphylococcus aureus, 79.1% and 75.0% of the isolates being methicillin ressitant Staphylococcus epdermidis (MRSE) or methicillin-ressitant Staphylococcus aureus (MRSA), respectively. No vancomycin intermediate or resistant staphylococcus was detected, but 4.9% teicoplanin intermediate rate was found in Staphylococcus epdermidis. The resistant rates to quinolones increased with the patient age. CONCLUSIONS: Bacterial resistance in UTI in Chinese females is a severe problem, which could be resulted from antimicrobial irrational use. In the selection of antimicrobial agents in the treatment of UTI, doctors should refer to the results of bacterial resistant surveillance. PMID- 19563061 TI - [Efficacy and safety of drospirenone-ethinylestradiol on contraception in healthy Chinese women: a multicenter randomized controlled trial]. AB - OBJECTIVE: To evaluate the contraception efficacy, mode of bleeding, side effects and other positive effects of drospirenone-ethinylestradiol (Yasmin) in healthy Chinese women. METHODS: This was a multicenter, randomized, control study of 768 healthy Chinese women who consulted about contraception. The subjects were randomized into Yasmin group (30 microg ethinylestradiol plus 3 mg drospirenone, 573 cases) or desogestrel group (30 microg ethinylestradiol plus 150 microg desogestrel, 195 cases) with the ratio of 3:1. Each individual was treated for 13 cycles. Further visits were required at cycle 4, cycle 7, cycle 10 and cycle 13 of treatment Weight, height, body mass index were evaluated at each visit. The menstrual distress questionnaire (MDQ) was given to the women at baseline, visit 3 (cycle 7) and visit 5 (after cycle 13). RESULTS: The values of basal features were similar between two groups (P > 0.05). The Pearl index (method failure) of Yasmin was 0. 208/hundred women year which was lower than that of desogestrel (0. 601/hundred women year). The mode of bleeding was similar between two groups after trial without showing any significant difference. According to MDQ subscale, the improvement of water retention and increasing appetite during inter menstrual period and water retention and general well-being during menstrual period in the Yasmin group (-0.297, -0.057, 0.033, 0.150 respectively) was more obvious than that in the desogestrel group (-0.108, 0.023, 0.231, -0.023 respectively) with a significant difference (P < 0.05). Some other values which improved in both two groups, especially the improvement of breast tenderness and pain and skin abnormality in Yasmin group (18.0%, 89/494; 12.6%, 62/494) was more distinct than that in desogestrel group (11.3%, 19/168; 5.4%, 9/168). The mean weight increased in desogestrel group (0.57 kg) while it decreased in Yasmin group (-0.28 kg) with a significant difference (P < 0.01). CONCLUSIONS: Both Yasmin and desogestrel have good efficacy on contraception and similar modes of menstrual bleeding. Yasmin is better than desogestrel in terms of weight control and premenstrual syndrome of oral contraceptive. PMID- 19563062 TI - [Effect of gonadotropin-releasing hormone-I agonist and gonadotropin-releasing hormone-II on endometrial carcinoma cell lines with different states of PTEN]. AB - OBJECTIVE: To study the effect of gonadotropin-releasing hormone-I (GnRH-I) agonist triptorelin and gonadotropin-releasing hormone-II (GnRH-II) on human endometrial carcinoma with different states of PTEN. METHODS: The endometrial carcinoma cells (Ishikawa, Ishikawa-PTEN, and Ishikawa-neo) were treated with different concentrations of triptorelin (10(-11) to 10(-5) mol/L) or GnRH-II (10( 11) to 10(-5) mol/L). Thirty min later, serine/threonine protein kinase (Akt) and extracellular signal-regulated kinase (ERK) 1/2 activation were detected using western blot method. 48 h later, the cell proliferation, cell cycle and apoptosis were detected using methyl thiazolyl tetrazolium (MTT) and flow cytometry. 17beta estradiol(17beta-E2, 10(-8) mol/L) or the specific estrogen receptor (ER) antagonist, ICI182780I (10(-6) mol/L) was added. After using the two drugs: triptorelin or GnRH-II, the above parameters were detected again. RESULTS: After treated with different concentrations (10(-11), 10(-9), 10(-7), 10(-5) mol/L) of triptorelin and GnRH-II, the cell growth was slowed, the percentage of G0/G1 phase cells increased, the percentage of G2/M and S phase cells decreased and the apoptosis rate increased in a dose-dependent manner (P < 0.01, P < 0.05). These changes were more obvious in Ishikawa. The apoptosis rate induced by GnRH-II was higher than that by the same concentration of triptorelin in the three cell lines. Triptorelin and GnRH-II inhibited the Akt and ERK1/2 activity in the endometrial carcinoma cells. 17beta-E2 counteracted the effect of triptorelin and GnRH-II on the endometrial carcinoma cells (P < 0.01, P < 0.05). CONCLUSION: Triptorelin and GnRH-II can promote apoptosis rate of endometrial carcinoma cells and inhibit cell proliferation in a dose-dependent manner which may be caused by ERK1/2 and Akt activity inhibition, and is related to the status of PTEN and could be offset by 17beta-E2. PMID- 19563063 TI - [Expression of connexin 43 in ovarian cancer and its relationship with chemoresistance]. AB - OBJECTIVE: To examine the expression of protein kinase C (PKC), connexin 43 (Cx43) and non-phosphorylated Cx43 in ovarian cancer, and discuss the role of phosphorylated of Cx43 in chemoresistance in ovarian cancer. METHODS: We examined the expression of Cx43, non-phosphorylated Cx43 and PKC in ovarian cancer tissue by immunohistochemistry, and compared their expression in chemosensitivity group and chemoresistance group. Cisplatin resistant ovarian cancer cell line SKOV3/DDP cells were treated by staurosporine (a kind of PKC inhibitors). Then expression of Cx43, nonphosphorylated Cx43 and PKC were tested. Meanwhile, we tested chemosensitivity of SKOV3/DDP cells by ATP bioluminescence tumor chemosensitivity assay (ATP-TCA). RESULTS: (1) Immunohistochemically, the rates of positive expression of Cx43 and non-phosphorylated Cx43 were 54%, 14% respectively in the chemoresistance group, which were 83%, 59% in the chemosensitivity group respectively (P < 0.05). The rate of positive expression of PKC in 28 chemoresistance ovarian cancer cases (64%) was higher than that in 29 chemosensitivity cases (31%, P < 0.05). Both of them were significantly lower in chemoresistance group than in chemosensitivity group (P < 0.05). In addition, the expression of PKC was negatively correlated with the expression of Cx43 and non phosphorylated Cx43. The correlation coefficients were -0.626 and -0.714, respectively (P < 0.05). (2) Immunohistochemically, PKC was down regulated, and Cx43 and non-phosphorylated Cx43 were up regulated in SKOV3/DDP cells after staurosporine treatment. The longer the staurosporine worked, the more expression of Cx43 was. (3) By ATP-TCA, SKOV3/DDP cells were resistant to paclitaxel and cisplatin. The tumor growth inhibition was higher in the group of paclitaxel or cisplatin combined staurosporine than in the group of paclitaxel or cisplatin alone. The sensitivity was intermediate in the group combined with low concentration staurosporine (1 x 10(-8) mol/L), and the sensitivity was high in the group combined with high concentration staurosporine (1 x 10(-7) mol/L). CONCLUSIONS: Phosphorylation of Cx43 caused by PKC leads to decrease in the expression of Cx43. This effect makes ovarian cancer cells less chemosensitive. Phosphorylation of Cx43 caused by PKC can be inhibited by staurosporine. PMID- 19563064 TI - [CDH1 polymorphism and its association with the risk of cervical cancer]. AB - OBJECTIVE: To investigate the effect of CDH1 3'-UTR + 54C/T single nucleotide polymorphism (SNP) on expression of luciferase reporter gene and its association with susceptibility to cervical cancer. METHODS: The luciferase gene expression vectors containing CDH1 3'-UTR+54C/T SNP C or T allelotype were constructed. The effect of CDH1 3'-UTR+54C/T SNP on expression of luciferase reporter gene in 293 T cells were tested by daul luciferase reporter assay system. The CDH1 3' UTR+54C/T SNP was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 280 cervical cancer patients and 330 healthy controls. RESULTS: The luciferase activity analysis showed that the relative luciferase activity (RLA) of 293T cells with C allelotype was 1.46, which was significantly lower than that of the 293 T cells with T allelotype (3.01; t = 2.94, P = 0.042). There was no significant difference in age distribution between the cervical cancer patients and the healthy controls. The genotype frequency distribution of CDH1 3'-UTR+54C/T SNP in healthy controls did not significantly differ from that expected by Hardy-Weinberg equilibrium (P > 0.05). The C allelotype frequency of CDH1 in cervical cancer patients was 80.%, which was significantly higher than that in healthy controls (74.5%; chi2 = 6.59, P = 0.010). The T/T, T/C and C/C genotype frequencies of cervical cancer patients and healthy controls were 4.3%, 30.0%, 65.7% and 5.8%, 39.4%, 54.8%, respectively, which were significantly different (chi2 = 7.45, P = 0.024). Compared with individuals with T/T or T/C genotype, individuals with C/C genotype had significantly higher risks of developing cervical cancer (OR = 1.578, 95% CI = 1.136 - 2.191). CONCLUSION: The C allelotype of CDH1 3'-UTR + 54C/T SNP might decrease the expression of luciferase reporter gene and the C/C genotype might be a potential risk for cervical cancer development PMID- 19563065 TI - [Some problems in the revision hip arthroplasties]. PMID- 19563066 TI - [Analysis of the failure mechanisms of the revision hip arthroplasties]. AB - OBJECTIVE: To investigate the failure mechanisms of revision hip arthroplasties and evaluate the effects of surgical technique, prosthesis design and patient related risk factors on different failure mechanisms. METHODS: A review of all revision hip arthroplasties from November 1995 to June 2008 identified 30 patients who underwent 30 revisions with 18 males and 12 females. The overall mean age for primary arthroplasties was 49 years (range 25-68 years) and 53 years (range 27-72 years) for index revision arthroplasties and the average interval between these two operations was 43.8 months (0-156 months). The failure mechanisms of index revision arthroplasties and primary arthroplasties were assessed and compared. Direct comparisons were made of data for the different age categories in terms of time to failures and reasons for failures. RESULTS: Regarding revision or prosthesis removal as the end point of the study, the reasons for 30 revision arthroplasties were aseptic loosening in 22 hips (73.3%), infection in 4 hips (13.3%), periprosthetic fracture in 3 hips and instability in 1 hip (3.3%). The overall mean age for last arthroplasties or prosthesis removal was 58 years (range 38-77 years) with an average interval of 78.8 months (range 1 216 months) from previous revision arthroplasties. The mean time to failure for patients above 60 years of age was significantly shorter than patients below 60 years of age (P < 0.01). CONCLUSION: The majority of failure mechanisms of revision hip arthroplasties are ineffective fixation of revisional implants and recurrence of local infection, which reveals the limitations to joint reconstruction philosophy and surgical technique. PMID- 19563067 TI - [An investigation on failure mechanisms of primary 327 hip arthroplasties]. AB - OBJECTIVE: To summarize the detailed failure mechanisms of revision hip arthroplasties and related risk factors. METHODS: From November 1988 to July 2008 revision of total hip arthroplasties was performed in 327 patients. The medical history, clinical and imaging material and operation records were investigated. RESULTS: Regarding revision as the end point of the study, the reasons for 327 revision arthroplasties were aseptic loosening in 226 hips (69.1%), infection in 52 hips (15.9%), periprosthetic fracture in 22 hips (6.7%), instability in 17 hips (5.2%), stem fracture in 5 hips (1.5%) and liner dissociation in 5 hips (1.5%). CONCLUSIONS: The main failure mechanisms of primary hip arthroplasties are aseptic loosening and infection of implants, which could be attributed to improper selection of operation indications and implants and limitations to surgical philosophy and technique. PMID- 19563068 TI - [Reconstruction of the severe acetabular bone defect by using metal mesh and impaction bone grafting technique]. AB - OBJECTIVES: To evaluate the medium-term clinical results of reconstruction of the severe acetabular bone defect by using metal mesh and impaction bone grafting (IBG) technique, as well as to emphasize the importance of Paprosky acetabular bone defect classification system in assessing the severity of bone defect and to analyse the failure reasons. METHODS: Between December 1998 and December 2007, 67 total hip arthroplasty (THA) revisions were made by using IBG technique to reconstruct severe acetabular bone defects combining with metal mesh or meta mesh cup on 63 patients. All the defects were combined defect (AAOS Type 3). There were 20 Paprosky II B defects in 19 patients, 28 Paprosky II C defects in 29 patients and 13 Paprosky II A defects in 12 patients. Regular follow-ups, involving the assessments of Harris hip scoring system, clinical efficacy, imaging and complications, were subsequently made. RESULTS: Sixty-one hips in 58 patients gained an average of 63 months (8-107) follow-up. Harris hip score increased from an average of 41.7 points (21-52) preoperatively to an average of 89.2 points (81-98) at the last follow-up, with an excellent and good rate of 93%. Radiographically, there were no loosening cases excluding the 3 dislocated polyethylene cups from the metal mesh cups. One case was failed to reconstruction the height of normal hip center, in which metal mesh cup was used for enforce the medial wall. Dislocations occurred in 3 hips, 1 of these patients required an open reduction and the other 2 dislocations only need close reduction. Postoperative infection rate was 1.6% (1 case), two stage revision with another IBG procedure succeeded in this patient. CONCLUSIONS: IBG combing with metal mesh for reconstruction of severe acetabular bone defect is an ideal technology. Paprosky acetabular bone defect classification system is very important in IBG procedure besides AAOS acetabular bone defect classification system to evaluate the severity of bone defect and to compare the outcomes between different authors. The use of metal mesh cup should be avoided to enforce acetabular medial wall in patients with severe acetabular bone defect. PMID- 19563069 TI - [Extended trochanteric osteotomy for removal of solidly-fixed femoral implants in total hip revision]. AB - OBJECTIVE: To analyze the clinical and radiographic results of the extended trochanteric osteotomy in revision total hip arthroplasty procedure. METHODS: The data of 27 patients (27 hips) of the clinical and radiographic results were reviewed between January 1998 and June 2005. There were 16 men and 11 women, and the mean age at the time of revision surgery was 63 years (range 42 to 78 years). Indications for use of the trochanteric osteotomy were removal of well-fixed femoral implants. Clinical evaluation included Harris and WOMAC scores. The radiological evaluation included stem subsidence, trochanteric fragment fractures and stem position change. RESULTS: Nineteen patients (19 hips) were followed up for an average of 5.3 years. All osteotomy sites healed by 16 months,with an average time to union of 16 weeks. No intraoperative fracture at the osteotomy site occurred. There was only one dislocation postoperatively. Three femoral components were subsidence at a mean of 3.4 mm (range 3-7 mm) in the first 6 weeks postoperatively; It united within 6 months without further migration. CONCLUSIONS: The extended trochanteric osteotomy facilitates the removal of well fixed femoral implants, allows reliable reattachment of the trochanteric fragment and results an excellent rate of healing and implant stability in revision total hip arthroplasty during mid-term follow-up. Long-term follow-up is needed. PMID- 19563070 TI - [The use of uncemented extensively porous-coated femoral components in the management of Vancouver type B2 periprosthetic femoral fractures]. AB - OBJECTIVE: To assess the outcome of patients with Vancouver type B2 periprosthetic fractures treated with femoral revision using an uncemented extensively porous-coated implant. METHODS: From October 2002 to May 2007, a retrospective clinical and radiographic assessment of 10 patients with Vancouver type B2 periprosthetic fractures treated with femoral revision was evaluated, among which 4 was cement fixation, 6 was non-cement fixation in the index procedure. RESULTS: The mean follow-up for these patients was 44 months (range, 12 to 67 months). The mean Harris hip score was 87.6 points. All 10 reconstructions showed solid fracture union and a stable prosthesis, 8 was bone ingrowth, 2 was fibrous ingrowth. Complications in 4 patients included thigh pain in 1, stress shield in 3. CONCLUSIONS: Vancouver type B2 periprosthetic fractures treated with uncemented extensively porous-coated femoral stems incorporate distally allowing stable fixation. PMID- 19563071 TI - [Study on visualized virtual surgery of living-related donor liver transplantation]. AB - OBJECTIVE: To investigate the significance of three dimensional visualization and virtual surgery system in living related donor liver transplantation surgery. METHODS: Two patients suffered biliary calculi were scanned by 64 slice helical computer tomography (CT) on livers and the data were imported into medical image proceeding system (MIPS) for sequence. Man-made segmentation and true-up on the image from the data were carried out. Three dimensional (3D) models of the liver and the intrahepatic vessels were reconstructed by VTK software respectively. The models were exported with format STL from it and then were imported into the FreeForm Modeling System for smoothing and modifying. At last, living related donor liver transplantation were simulated with the force-feedback equipment (PHANToM). RESULTS: It had great verisimilar image for the reconstructed 3D liver models with artery, hepatic vein, portal vein and bile duct. By seeing through liver, it had high fidelity and strong 3D effect for the intrahepatic artery, hepatic vein, portal vein and bile duct, and their spatial disposition and course and co-relationship were shown clearly. In the virtual surgery system, the virtual scalpel could be manipulated on 3D liver model with PHANToM. The simulating effect was the same as the clinic operation for living related donor liver transplantation. CONCLUSIONS: The visualized liver model reconstructed is 3D and verisimilar, and it is helpful to design reasonable scheme for liver transplantation. It can improve the surgical effect, decrease the surgical risk, reduce the complication, enhance the communication between doctor and patient through designing surgical plan and demonstrating visualized operation before surgery. PMID- 19563072 TI - [Study on the operation score system for endoscopic thyroidectomy]. AB - OBJECTIVE: To investigate the clinical significance of the operation score system for endoscopic thyroidectomy. METHODS: An operation score system based on 6 important procedure skills of endoscopic thyroidectomy was established. And a retrospective study of the first 300 consecutive patients underwent endoscopic thyroidectomy from July 2001 to December 2007 by a single surgeon was performed. The patients was divided into 10 consecutive groups chronologically, each comprising 30 cases. RESULTS: The mean operation score of all the patients was 6.0 and the mean operation time was 98.1 min. There were significant differences in the mean operation score, every skill score and the mean operation time among the 10 groups. In the consecutive two groups comparison, significant differences in the operation scores were observed between group 2 and 3 (P < 0.05) and between group 5 and 6 (P < 0.05). CONCLUSION: The operation score system for endoscopy thyroidectomy is a useful method to judge the proficiency and the stability of the operation. PMID- 19563073 TI - [The biomechanical study and clinical application of monosegmental pedicle instrumentation by endplate method in the fracture vertebrae for thoracolumbar fracture]. AB - OBJECTIVE: To evaluate the biomechanical and clinical effect of the treatment of thoracolumbar fracture with monosegmental pedicle instrumentation in the fracture vertebrae by endplate method. METHODS: Twenty-four porcine thoracolumbal spinal model, divided into four groups randomly, compared the stability of these four groups through pull-out testing. Retrospective study of 49 patients with thoracolumbar fracture who were treated with this technique, to observe the fusion of bone graft, the height of the anterior and posterior range, the angle of kyphosis and the volume of spinal canal, the loss of rectification, low back pain, and the limitation of activity of lumbar. RESULTS: The endplate method group is more stability than the parallel method group, and the pedicle screw in the fracture vertebrae can get enough stability. After operation all 49 cases achieved satisfactory reduction, strong bone fusion, no reduction loss, no refractoriness low back pain, limitation of motion of lumbar et al. CONCLUSION: Monosegmental pedicle instrumentation by endplate method in the fracture vertebrae for thoracolumbar fracture can get enough extraction stability, and get satisfied clinical effect. PMID- 19563074 TI - [Experimental study on spinal fusion induced by hBMP-4 gene modified tissue engineered bone]. AB - OBJECTIVE: To evaluate the efficacy of hBMP-4 gene modified tissue engineered bone graft in the enhancement of rabbit spinal fusion and find an ideal kind of substitute for the autograft bone. METHODS: Rabbit BMSCs were cultured and transfected with AAV-hBMP-4 using different MOI value. The optimal MOI value were determined by observing cell's morphology change. BMSCs were then transfected with AAV-hBMP4 and AAV-EGFP respectively, following which the transfected cells were evenly suspended in a collagen sponge I, and implanted to either side of the L5,6 intertransverse spaces posterolateral in the New Zealand rabbits to induce spinal fusion. Fourteen rabbits were randomly divided into 2 groups. Group 1: AAV hBMP-4 transfected BMSCs in the right side (hBMP-4 side) and autograft bone in the left side. Group 2: AAV-hBMP-4 transfected BMSCs in the right side (hBMP-4 side) and AAV-EGFP transfected BMSCs in the left side (EGFP side). Radiographs and three-dimensional CT of the spine, manual palpation, gross and histological examination of the fusion masses for all the animals were performed subsequent to animals having been sacrificed at 12 weeks after surgery. RESULTS: Evaluation has been taken in 12 New Zealand rabbits delivered into 2 groups which meet the criterion after operation. Eleven in 12 implemented sides involved hBMP-4 achieved bony fusion, to which 5 in 6 autografted sides was similar. But only 2 in 6 sides in EGFP-group achieved bony fusion meanwhile. Three-dimensional CT scan and palpation also evidenced the results. Bone formation was observed obviously on specimen both in hBMP4 sides and autografted ones. EGFP-group also got bony integration, but the quantity was small. CONCLUSION: Tissue-engineered bone graft constructed from application of hBMP4 is a fine substitute for autograft. Effective enhancement of bony integration in spinal fusion surgery has been evidenced in vivo. PMID- 19563075 TI - [Expression, purification of tetanus toxin C fragment/cardiotrophin-1 recombinant fusion protein, target delivery to CNS and neurotrophy biology ability]. AB - OBJECTIVE: Expression, purification of tetanus toxin C fragment/cardiotrophin-1 recombinant fusion protein (CT-1/TTC) in BL21 (DE3) E. coli, examined whether tetanus toxin C fragment mediate the cardiotrophin-1 target delivery to the central nervous system and the cardiotrophin-1 has the neurotrophic ability. METHODS: Induction by IPTG, the fusion protein was expressed and then purified by GST affinity agarose. The interest protein was viewed by SDS-PAGE, further characterized by Western Blot Rat sciatic nerve transected model was selected. Using drug by nerve-regeneration-chamber and intramuscular injection. Execute these animals one week after the operation. The L4-L6 segments of the spinal cord were harvested after transaortic perfusion with 4% paraformaldehyde. The freeze sections of spinal tissues were stained with immunohistochemistry method. And select the new born SD rat sciatic nerve transected model, using CT-1/TTC fusion protein by muscle injection. Execute these animals one week after the operation. The L4-L6 segments of the spinal cord were harvested after transaortic perfusion with 4% paraformaldehyde. The freeze sections of spinal tissues were stained by Nissl's staining. RESULTS: After induction, the fusion protein was about 15% of the total protein and the soluble part was predominant. Purified by GST fusion protein column, the interest protein's concentration is 2.7 g/L. The CT-1/TTC fusion protein was found in lumbar intumescentia by immunohistochemistry method. And after sciatic nerve transected, the numbers of cornu anterius medullae spinalis motoneurons in L4-L6 segments, compared to CT-1/TTC protein grope, have a lower survival rate. CONCLUSIONS: The recombinant CT-1/TTC protein can be expressed and purified in BL21 (DE3) E. coli. This fusion protein has two biological activities of targeting delivery to central nervous system and protecting the cornu anterius medullae spinalis motoneurons. PMID- 19563076 TI - [Expression of hepatocyte growth factor, transcription factor SP1, vascular endothelial growth factor and CD34 in serosa-infiltrative gastric cancer and their relations with defluxion of gastric cancer cell and prognosis]. AB - OBJECTIVE: To observe the expression of hepatocyte growth factor (HGF), transcription factor SP1, vascular endothelial growth factor (VEGF) and CD34 (demonstrating by microvessel density, MVD) in serosa-infiltrative gastric cancer (T3) and their relations with the pathobiological behavior of the tumor, and to investigate the molecular basis of the defluxion of gastric cancer cells in abdominal cavity and its influence on prognosis. METHODS: Selective collection of peritoneal lavage was obtained from 80 patients with serosa-infiltrative gastric cancer received operation from April to December in 2007. The cancer cells were detected by using peritoneal lavage cytology (PLC) and immunochemistry of cytokeratin 18 (CK18). Immunohistochemistry was applied to detect the HGF, SP1, VEGF and CD34 in serosa-infiltrative gastric cancer tissues. The rigorous follow up was carried out for the patients. RESULTS: The positive rate of PLC was 63.8% (51/80), and the positive rate of immunochemistry of CK18 was 75.0% (60/80). The positive cases in PLC were positive in immunochemistry of CK18 also, while 9 negative cases in PLC were positive with CK18, and of them 6 cases were determined positive with exfoliated cancer cells through pathological consulting. So the positive rate of exfoliated cells of this group was 71.3% (57/80). The positive rates of HGF, SP1 and VEGF in gastric cancer tissues were 57.5%, 52.5% and 55.0%, respectively, and were all significantly correlated with the MVD (P < 0.05). HGF, SP1, VEGF and MVD were correlated with the positive rate of exfoliated cells (P < 0.05). HGF, SP1, VEGF and MVD were found significantly related to prognosis on univariate analysis (P < 0.05), and it was demonstrated that HGF, SP1 and VEGF were independent prognostic influential factors on Logistic regression analysis (P < 0.05). CONCLUSIONS: The expression of HGF, SP1, VEGF and MVD are related with the biological behaviour of serosa-infiltrative gastric cancer. The detection of these factors might be helpful in predicting the defluxion of gastric cancer cells and postoperative recurrence. PMID- 19563077 TI - [Effect of stromal cell-derived factor-1 and its receptor CXCR4 on liver metastasis of human colon cancer]. AB - OBJECTIVE: To investigate the effect of chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 on liver metastasis of human colon cancer. METHODS: Expression of CXCR4 in different colon cancer cell lines and SDF-1 in different tissues were detected by using Western-blot technique. Effect of SDF-1 and anti-CXCR4 monoclonal antibody (McAb) on proliferation and migration of HT-29 cells were measured using MTT methods. Model mimicking liver metastasis of human colon cancer was established by injecting HT-29 cells intrasplenically into BALB/C nude mice. Mice were randomly divided into AMD3100 treated group and control group. Liver metastatic rate and tumor foci were measured 7 weeks after. RESULTS: HT-29 cells expressed higher level of CXCR4 protein, and liver tissue expressed higher level of SDF-1 protein. Compared with the control, SDF-1 could significantly induced the proliferation and migration of the HT-29 cells, and anti-CXCR4 McAb could inhibited both functions of SDF-1. The liver metastasis rate in the control group was 100%, and it was 40% in the AMD3100 treating group (P < 0.05). The mean liver metastasis number also significantly decreased by AMD3100 (7.8 +/- 2.6 vs 22.4 +/- 8.6, P < 0.05). CONCLUSIONS: SDF-1/CXCR4 biological axis play an important role in liver metastasis of human colon cancer. Arrest of CXCR4 can inhibit liver metastasis of colon cancer through blocking cell proliferation and migration induced by SDF-1. PMID- 19563078 TI - [Research of the relationship between insulin signal transduction and glucose transportation in rats after trauma]. AB - OBJECTIVE: To investigate the relationship between insulin post-receptor signal transduction and the change of glucose transportation by skeleton muscle after surgical trauma in rats. METHODS: Small intestine bowel resection was performed to establish the surgical trauma model in rats. The content and the phosphorylation state of two key proteins in the insulin signaling pathway: insulin receptor substrate-1 (IRS-1) and protein kinase B (PKB/Akt) in skeletal muscle were measured respectively. The 3H labeled glucose uptake experiment was carried out to evaluate the glucose transportation function in both groups. Finally, the expression and the distribution of glucose transporter 4 (GLUT4) in skeletal muscle were detected respectively. RESULTS: The total content of IRS-1 and PKB/Akt in skeletal muscle in both groups had no difference. The phosphorylation of tyrosine (Tyr) residue of IRS-1 in the operation group was attenuated by 31% (P = 0.018), whereas the phosphorylation of serine (Ser) residue of IRS-1 was significantly enhanced by 63% compared with the control group (P = 0.000). Accordingly, the phosphorylation state of PKB/Akt (activated) was attenuated by 48% in the operation group (P = 0.000). The rate of 2-Deoxy-D [1-3H] glucose transported by skeletal muscle in the operation group was significantly lower than that in the control group. Both the expressions of GLUT 4 mRNA and the total content of GLUT-4 protein in two groups had no significant difference. But the expression of GLUT4 in the plasma membrane was decreased in the operation group than that in the control group. CONCLUSIONS: Insulin resistance was associated with enhanced Ser phosphorylation of IRS-1, which impaired its interaction with its downstream target PKB/Akt. Such impaired interactions abolished the ability of IRS-1 to undergo insulin-induced Tyr phosphorylation and further propagate the insulin receptor signal. Uncoupling of signal transduction led to decrease in glucose uptake which associated with a defect in insulin-stimulated glucose transport and GLUT-4 translocation. PMID- 19563080 TI - A jaw for a tooth--the human rights cost of the Gaza invasion. PMID- 19563079 TI - [The effect of rosiglitazone on the activity of STAT1 in rats with severe acute pancreatitis]. AB - OBJECTIVE: To explore the effect of rosiglitazone on the activity of signal transducer and activator of transcription 1 in rats with severe acute pancreatitis. METHODS: Fifty-four male Wistar rats were randomly allocated into three groups (n = 18). SO group: sham-operated animals served as control, operation was executed and sodium chloride but not sodium taurocholate was injected. SAP group: SAP was induced by the retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. ROSI group: same as SAP group, but rosiglitazone (6 mg/kg) was administered intravenously 30 min before operation. Rats in each group were sacrificed at 3,6 and 12 h after operation. The levels of serum amylase and histologic scores of pancreatic tissue were measured. The expression of TNF-alpha mRNA in pancreatic tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR). The expression of phosphorylated STAT1 in pancreatic tissue was assayed by immunohistochemistry. RESULTS: Compared to SO group, the levels of serum amylase and phosphorylated STAT1, TNF-alpha mRNA and histologic scores of pancreatic tissue were significantly elevated at the same time points after SAP (P < 0.01). The levels of these detection in ROSI group were lower than those of the SAP group at the same time points (P < 0.05), but higher than SO group (P < 0.05). CONCLUSIONS: STAT1 was activated in severe acute pancreatitis. Rosiglitazone has a protective effects in rats with severe acute pancreatitis. The mechanism of its protective effects maybe that it inhibits the activation of JAK/STAT pathway, which can down regulate the expression of TNF-alpha mRNA and block the the inflammatory cascade partially. PMID- 19563081 TI - Misinformation and lack of knowledge hinder cervical cancer prevention. PMID- 19563082 TI - Knees, Comrades and sample size. PMID- 19563083 TI - 'Loose cannon' Letlape says he was 'misinterpreted'. PMID- 19563084 TI - Cholera--getting the basics right. PMID- 19563085 TI - 'Ethical' tariffs to be scrapped by May--HPCSA. PMID- 19563086 TI - Pitfalls of administering drugs via nasogastric tubes. PMID- 19563087 TI - Key facts on male circumcision. PMID- 19563088 TI - Intramyocardial tuberculosis--a rare underdiagnosed entity. PMID- 19563090 TI - Early detection of infant hearing loss in South Africa. PMID- 19563091 TI - Community assault--the cost of rough justice. PMID- 19563092 TI - From 'playstation thumb' to 'cellphone thumb': the new epidemic in teenagers. PMID- 19563093 TI - High rate of adverse events following circumcision of young male adults with the Tara KLamp technique: a randomised trial in South Africa. AB - BACKGROUND: The Tara KLamp (TK) device has been claimed to enable circumcisions to be performed safely and easily in medical and non-medical environments. Published evaluation studies have been conducted among young children only. METHODS: Following a randomised controlled trial (RCT) on 3 274 participants on the impact of male circumcision on HIV transmission, 69 control group members participated in this male circumcision methods trial and were randomised to a forceps-guided (FG) group and a TK group, and circumcised. RESULTS: Of the 166 men asked to participate, 97 declined, most (94) refusing circumcision by the TK technique; 34 men were randomised to the FG group and 35 to the TK group, and 32 and 24 patients were circumcised by the FG and TK methods respectively, of whom 29 and 19 respectively attended the post-circumcision visit. All 12 adverse event sheets corresponded to the TK group (p<0.001) and circumcisions by the TK method. Less favourable outcomes were associated with the TK method, including any sign of an adverse event (37% v. 3%; p=0.004), delayed wound healing (21% v. 3%; p=0.004) and problems with penis appearance (31% v. 3%; p=0.001). Participants randomised to the TK method were significantly more likely to report bleeding (21% v. 0%; p=0.02), injury to the penis (21% v. 0%; p=0.02), infection (32% v. 0%; p=0.002), swelling (83%/ v. 0%; p<0.001), and problems with urinating (16% v. 0%; p=0.056). The mean score of self-estimated pain was 9.5 for participants circumcised by TK compared with 6.1 for other participants (adjusted p=0.003). CONCLUSION: This study provides compelling evidence that strongly cautions against use of the TK method on young adults. PMID- 19563094 TI - Antimicrobial susceptibility of methicillin-resistant Staphylococcus aureus isolates from South Africa. AB - OBJECTIVE: Trends in the antibiotic susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) are regularly investigated in many countries, but minimal countrywide data are available for South Africa. The aim of this study was to describe the antibiotic susceptibility patterns of MRSA isolates collected in South Africa. DESIGN: Susceptibility testing of 248 MRSA isolates collected from 15 National Health Laboratory Services (NHLS) and 8 private laboratories against 17 antibiotics was performed using the disc diffusion method. Demographic data were collected and correlated with antibiotic resistance patterns. RESULTS: Antibiotic resistance of MRSA to erythromycin, tetracycline, trimethoprim/sulfamethoxazole, gentamicin and ciprofloxacin ranged between 55% and 78%, while all isolates were susceptible to teicoplanin, linezolid, vancomycin and quinopristin/dalfopristin. A significant difference in the resistance pattern of the isolates towards certain antimicrobial agents was identified among adults and children, as well as between isolates collected from the private and NHLS laboratories. CONCLUSION: This is the first extensive report on the antimicrobial susceptibilities of South African MRSA isolates. These data can assist treatment decisions and form a baseline for further surveillance. PMID- 19563095 TI - Assessing missed opportunities for the prevention of mother-to-child HIV transmission in an Eastern Cape local service area. AB - BACKGROUND: Prevention of new HIV infections is a critical imperative for South Africa; the prevention of mother-to-child transmission (PMTCT) is one of the most efficacious HIV prevention interventions. OBJECTIVE: Assessment of a PMTCT programme to determine missed opportunities. SETTING: The Kouga local service area (LSA), bordering Nelson Mandela Bay Municipality (Port Elizabeth) in the Eastern Cape. METHODS: An assessment was conducted in 2007 before implementing technical support for strengthening the PMTCT programme, including: interviews with 20 PMTCT managers, 4 maternity staff and 27 other health workers on service provision, management, infrastructure, human resources and the health information system; 296 antenatal clinic users on their service perceptions; 70 HIV-positive women on HIV knowledge, infant feeding, coping, support and service perceptions; 8 representatives from community organisations and 101 traditional health practitioners (THPs). Observations were conducted during site visits to health facilities, and the District Health Information System (DHIS) data were reviewed. RESULTS: Staff had high levels of awareness of HIV policies and most had received some relevant training. Nevirapine uptake varied by clinic, with an average of 56%. There were many missed opportunities for PMTCT, with 67% of pregnant women tested for HIV and only 43% of antenatal care attendees tested during a previous pregnancy. Only 6% of HIV-positive women reported support group participation. CONCLUSIONS: Reducing missed opportunities for PMTCT requires strengthening of the formal health sector, intersectoral liaison, and greater community support. Priority areas that require strengthening in the formal health sector include HIV counselling and testing; family planning and nutrition counselling; infant follow up; human resources; and monitoring and evaluation. PMID- 19563096 TI - Child's play: exposure to household pesticide use among children in rural, urban and informal areas of South Africa. AB - BACKGROUND: As part of a larger dermatological investigation undertaken in 1999 - 2001 involving the Department of Dermatology, Groote Schuur Hospital (Cape Town, South Africa) and Nottingham University (UK), household pesticide use was investigated among Xhosa-speaking families living in three areas in South Africa (a rural area, an urban township and an informal settlement). OBJECTIVES: The aim was to characterise pesticide use patterns and potential exposures through skin absorption, ingestion and inhalation for this group of South African children. METHODS: A standardised questionnaire, which included a section investigating household pesticide use, was administered by four trained fieldworkers to the parents/guardians of the 740 children (25%) aged between 3 and 11 years identified as having atopic dermatitis either by clinical examination or according to the UK criteria (rural N=387, urban N=292, informal N=61). RESULTS: Of the children with atopic dermatitis, 539 (73%) had been exposed to household pesticides. Most childhood exposure (89%) occurred in the informal settlements, followed by 78% in the urban area and 63% in the rural area. CONCLUSIONS: This research highlighted considerable home environment pesticide exposure of South African children in lower socio-economic groups in rural, urban and informal areas. As children are particularly vulnerable to the short- and long-term health effects of pesticide exposure, further in-depth investigation is needed to ascertain and document the health effects associated with such exposure in the home. PMID- 19563097 TI - Guideline for using growth hormone in paediatric patients in South Africa: treatment of growth hormone deficiency and other growth disorders. AB - The Paediatric and Adolescent Endocrine and Diabetes Society of South Africa (PAEDS-SA) recommends, in line with other international groups, that growth hormone (GH) therapy be considered for children and adolescents with significantly short stature and poor growth velocity in the following instances: GH deficiency; Turner syndrome; Prader-Willi syndrome; small-for-gestational-age children with failure of catch-up growth; idiopathic short stature; and chronic renal insufficiency. We have produced treatment guidelines for the use of GH, designed to allow flexibility to determine coverage on a case-by-case basis. We further recommend that when used for growth promotion, GH therapy should be initiated and monitored by, or in consultation with, a paediatric endocrinologist. PMID- 19563098 TI - DNA replication initiation. AB - DNA replication is fundamental to cellular life on earth, and replication initiation provides the primary point of control over this process. Replication initiation in all organisms involves the interaction of initiator proteins with one or more origins of replication in the DNA, with subsequent regulated assembly of two replisome complexes at each origin, melting of the DNA, and primed initiation of DNA synthesis on leading and lagging strands. Archaea and Eukarya share homologous systems for DNA replication initiation, but differ in the complexity of these; Bacteria appear to have analogous, rather than homologous, mechanisms for replication initiation. This chapter provides an overview of current knowledge of initiation of chromosomal DNA replication in the three domains of life. PMID- 19563099 TI - DNA replication fork proteins. AB - DNA replication is a complex mechanism that functions due to the co-ordinated interplay of several dozen protein factors. In the last few years, numerous studies suggested a tight implication of DNA replication factors in several DNA transaction events that maintain the integrity of the genome. Therefore, DNA replication fork proteins have also to be considered as part of a general process aiming at replicating and protecting the genome in order to allow the correct function of a cell and of its eventual daughter cells. This is illustrated by several DNA repair pathways such as base excision repair, nucleotide excision repair, double-strand break repair, and mismatch repair. Furthermore, several of the replication proteins have also been shown to be essential in sensing and transducing DNA damages through the checkpoint cascade pathways. This review will summarize the properties of DNA replication proteins that function exclusively at the replication fork. PMID- 19563100 TI - Random and site-specific replication termination. AB - Bi-directionality is a common feature observed for genomic replication for all three phylogenetic kingdoms: Eubacteria, Archaea, and Eukaryotes. A consequence of bi-directional replication, where the two replication forks initiated at an origin move away from each other, is that the replication termination will occur at positions away from the origin sequence(s). The replication termination processes are therefore physically and mechanistically dissociated from the replication initiation. The replication machinery is a highly processive complex that in short time copies huge numbers of bases while competing for the DNA substrate with histones, transcription factors, and other DNA-binding proteins. Importantly, the replication machinery generally wins out; meanwhile, when converging forks meet termination occurs, thus preventing over-replication and genetic instability. Very different scenarios for the replication termination processes have been described for the three phylogenetic kingdoms. In eubacterial genomes replication termination is site specific, while in archaea and eukaryotes termination is thought to occur randomly within zones where converging replication forks meet. However, a few site-specific replication barrier elements that mediate replication termination have been described in eukaryotes. This review gives an overview about what is known about replication termination, with a focus on these natural site-specific replication termination sites. PMID- 19563101 TI - Checkpoint regulation of DNA replication. AB - We discuss the mechanisms regulating entry into and progression through S phase in eukaryotic cells. Methods to study the G1/S transition are briefly reviewed and an overview of G1/S-checkpoints is given, with particular emphasis on fission yeast. Thereafter we discuss different aspects of the intra-S checkpoint and introduce the main molecular players and mechanisms. PMID- 19563102 TI - Introduction to molecular combing: genomics, DNA replication, and cancer. AB - The sequencing of the human genome inaugurated a new era in both fundamental and applied genetics. At the same time, the emergence of new technologies for probing the genome has transformed the field of pharmaco-genetics and made personalized genomic profiling and high-throughput screening of new therapeutic agents all but a matter of routine. One of these technologies, molecular combing, has served to bridge the technical gap between the examination of gross chromosomal abnormalities and sequence-specific alterations. Molecular combing provides a new perspective on the structure and dynamics of the human genome at the whole genome and sub-chromosomal levels with a resolution ranging from a few kilobases up to a megabase and more. Originally developed to study genetic rearrangements and to map genes for positional cloning, recent advances have extended the spectrum of its applications to studying the real-time dynamics of the replication of the genome. Understanding how the genome is replicated is essential for elucidating the mechanisms that both maintain genome integrity and result in the instabilities leading to human genetic disease and cancer. In the following, we will examine recent discoveries and advances due to the application of molecular combing to new areas of research in the fields of molecular cytogenetics and cancer genomics. PMID- 19563103 TI - Replication initiation point mapping: approach and implications. AB - Duplication of eukaryotic chromosomes begins from multiple sites called origins of replication, with DNA synthesis proceeding bidirectionally away from the origin. There is little detailed information available pertaining to whether replication initiates at specific sites or anywhere within a given origin. The development of replication initiation point (RIP) mapping has made it possible to map start sites for DNA synthesis at the nucleotide level. The key step in RIP mapping is the purification of nascent DNA, which is initiated by small RNA primers. For the removal of broken DNA fragments, we utilize lambda-exonuclease, which digests DNA, but leaves nascent strands intact as long as they have the RNA primer still attached. RIP mapping is a sensitive technique and has been successfully applied to single copy loci in both budding and fission yeast, archaebacteria, and human cells. Studies in yeast have shown that the binding site for the initiator, the origin recognition complex (ORC), lies immediately adjacent to the replication start point, which suggests that ORC directs the initiation machinery to a distinct site. Here, we present a detailed step-by-step protocol for RIP mapping of replication origins in budding yeast. PMID- 19563104 TI - Purification of restriction fragments containing replication intermediates from complex genomes for 2-D gel analysis. AB - In order to perform 2-D gel analyses on restriction fragments from higher eukaryotic genomes, it is necessary to remove most of the linear, nonreplicating, fragments from the starting DNA preparation. This is so because the replication intermediates in a single-copy locus constitute such a minute fraction of all of the restriction fragments in a standard DNA preparation - whether isolated from synchronized or asynchronous cultures. Furthermore, the very long DNA strands that characterize higher eukaryotic genomes are inordinately subject to branch migration and shear. We have developed a method that results in significant enrichment of replicating fragments that largely maintain their branched intermediates. The method depends upon two important factors: (1) replicating fragments in higher eukaryotic nuclei appear to be attached to the nuclear matrix in a supercoiled fashion, and (2) partially single-stranded fragments (e.g., those containing replication forks) are selectively adsorbed to benzoylated napthoylated DEAE (BND)-cellulose in high salt conCentrations. By combining matrix-enrichment and BND-cellulose chromatography, it is possible to obtain preparations that are enriched as much as 200-fold over the starting genomic DNA and are thus suitable for analysis on 2-D gels. PMID- 19563106 TI - Analysis of telomeric DNA replication using neutral-alkaline two-dimensional gel electrophoresis. AB - DNA replication studies often rely on analysis of replication intermediates, such as progressing replication forks and growing nascent strands. The assay presented here for replication at telomeres in the yeast Saccharomyces cerevisiae is based on the analysis of nascent DNA strands prior to the ligation step. Preligation replication intermediates are very rare due to their transient nature. To enrich for such intermediates, inhibition of the ligation step is performed by using a temperature-sensitive allele of the replicative ligase Cdc9 at nonpermissive temperature. The method can be used for fine analysis within rather short DNA fragments which makes it particularly advantageous for studying telomere replication. It can also be helpful for analysis of DNA recombination and potentially any process which involves ligation of nicked DNA. PMID- 19563105 TI - Topological analysis of plasmid DNA replication intermediates using two dimensional agarose gels. AB - A fundamental process in DNA replication is the disentangling of the two parental strands by DNA topoisomerases. In this chapter, I detail the topological analysis of plasmid replication intermediates using two-dimensional (2D) agarose gels. The method can resolve replication intermediates according to mass and topology, and can resolve unlinked monomeric circles from catenated dimers of varying topology. The method has been used, alone or in combination with a procedure for purifying covalent protein-DNA complexes, to analyse the effect oftopoisomerase inhibitors on the topology of replication intermediates, to map the location of drug stabilized topoisomerase cleavage complexes with respect to replication forks and to detect the breakage and repair of replication forks following collision with cleavage complexes. Other applications include the detection of knots that form independently of, or concomitantly with, DNA replication. PMID- 19563107 TI - Chromatin immunoprecipitation of replication factors moving with the replication fork. AB - Replication of chromosomes involves a variety of replication proteins including DNA polymerases, DNA helicases, and other accessory factors. Many of these proteins are known to localize at replication forks and travel with them as components of the replisome complex. Other proteins do not move with replication forks but still play an essential role in DNA replication. Therefore, in order to understand the mechanisms of DNA replication and its controls, it is important to examine localization of each replication factor. Here we describe a chromatin immunoprecipitation (ChIP) method to locate a replication factor at the replication fork. Defining the localization of replication proteins should provide important insight into mechanistic understanding of the regulation of the DNA replication process. PMID- 19563108 TI - Density transfer as a method to analyze the progression of DNA replication forks. AB - The density transfer technique is a valuable tool to examine the progression of individual DNA replication forks. It is based on the transfer of cells from a medium containing dense isotopes to a medium with light (normal) isotopes (or vice versa), to obtain DNA sequences hybrid in density that can be identified as replicated molecules. Using specific DNA probes along a chromosome, the dense isotope transfer method allows determining the extent of replication at any position of a replicon and the rate of replication fork progression. In the eukaryotic model budding yeast, this technique has been useful to establish a role for different proteins during the elongation of chromosomal replication and to analyze the movement and stability of DNA replication forks under different experimental conditions. PMID- 19563109 TI - High-resolution mapping of points of site-specific replication stalling. AB - Genetic instability due to stalled replication forks is thought to underlie a number of human diseases, such as premature ageing and cancer susceptibility syndromes. In addition, site-specific stalling occurs at some genetic loci. A detailed understanding of the topology of the stalled replication fork gives a valuable insight into the causes and mechanisms of replication stalling. The method described here allows mapping of the position of the 3'-end of the nascent leading or lagging strand at the replication fork, stalled at a site-specific barrier. The replicating DNA is purified, digested with restriction enzymes, and enriched by BND-cellulose chromatography. The DNA is separated on a sequencing gel, transferred to a membrane, and hybridised to a strand-specific probe. The data obtained using this method allow determining the position of the 3'-end of the nascent strand at a stalled fork with a one-nucleotide resolution. PMID- 19563110 TI - DNA replication in nucleus-free Xenopus egg extracts. AB - Extracts derived from Xenopus laevis eggs represent a powerful cell-free system to study eukaryotic DNA replication. A variation of the system allows for DNA replication not only in a cell-free environment, but also in the absence of a nucleus. In this nucleus-free system, DNA templates are licensed with High-Speed Supernatant (HSS) and then replicated with a concentrated NucleoPlasmic Extract (NPE). This method has the advantage of allowing replication of small plasmids with desired modifications and manipulation of the nuclear environment. This chapter describes the protocols needed to prepare HSS and NPE and how these extracts are used to study DNA replication. PMID- 19563111 TI - ChIP-chip to analyze the binding of replication proteins to chromatin using oligonucleotide DNA microarrays. AB - Chromatin immunoprecipitation (ChIP) is a widely used method to study the interactions between proteins and discrete chromosomal loci in vivo. Originally, ChIP was developed for analysis of protein associations with DNA sequences known or suspected to bind the protein of interest. The advent of DNA microarrays has enabled the identification of all DNA sequences enriched by ChIP, providing a genomic view of protein binding. This powerful approach, termed ChIP-chip, is broadly applicable and has been particularly valuable in DNA replication studies to map replication origins in Saccharomyces cerevisiae based on the association of replication proteins with these chromosomal elements. We present a detailed ChIP-chip protocol for S. cerevisiae that uses oligonucleotide DNA microarrays printed on polylysine-coated glass slides and can also be easily adapted for commercially available high-density tiling microarrays from NimbleGen. We also outline general protocols for data analysis; however, microarray data analyses usually must be tailored specifically for individual studies, depending on experimental design, microarray format, and data quality. PMID- 19563112 TI - Analyzing origin activation patterns by copy number change experiments. AB - Advances in microarray technology have enabled the analysis of replication dynamics on a genome-wide scale, providing deeper insight to the factors that regulate DNA replication. Studies using high-density microarrays have led to the genome-wide identification of replication origins in the budding yeast, Saccharomyces cerevisiae, and enabled the analysis of the global temporal pattern of origin activation under various conditions. We have developed a replication origin array that contains the approximately 430 potential origins in the yeast genome. By detecting the copy number change that occurs as cells progress from G1 to S phase on these arrays, we have produced origin activation patterns in wild type cells similar to those obtained from previous studies that used whole-genome arrays. We have also applied this method to study S phase checkpoint mutants, providing insight into the genome-wide regulation of replication origin activation by S phase checkpoint kinases in the presence of replication stress. The main procedures of this technique involve arresting yeast cells in G1 and S phase, isolating and labeling genomic DNA with fluorescent dyes, and cohybridizing the DNA samples to replication origin arrays to yield copy number change data. PMID- 19563113 TI - Detection of replication origins using comparative genomics and recombinational ARS assay. AB - Effective experimental techniques are available to identify replication origin regions in eukaryotic cells. Genome-wide identification of the precise sequence elements that direct origin activity is however still not straightforward, even in the yeast Saccharomyces cerevisiae which has the best characterised eukaryotic replication origins. The availability of genome sequences for a series of closely related (sensu stricto) budding yeasts has allowed us to take a 'comparative genomics' approach to this problem. Since they represent functional protein binding sites, origin sequences are conserved better than the surrounding intergenic sequence within the genomes of closely related yeasts. We describe here how phylogenetic comparison data can be used to identify candidate replication origin sequences in the S. cerevisiae genome, and how large numbers of such candidate sites can simultaneously be assayed for ability to initiate replication. Similar approaches could potentially be used to identify protein binding sequence elements having other functions, as well as replication origin sites in other species. PMID- 19563114 TI - Isolation of restriction fragments containing origins of replication from complex genomes. AB - The identification and isolation of origins of replication from mammalian genomes has been a demanding task owing to the great complexity of these genomes. However, two methods have been refined in recent years each of which allows significant enrichment of recently activated origins of replication from asynchronous cell cultures. In one of these, nascent strands are melted from the long template DNA, and the small, origin-centered strands are isolated on sucrose gradients. The second method involves the selective entrapment of bubble containing fragments in gelling agarose and their subsequent recovery and isolation by molecular cloning. Libraries prepared by this method from Chinese hamster and human cells have been shown to be extremely pure, and provide a renewable resource of origins that can be used as probes on microarrays or sequenced by high-throughput techniques to localize them within the genomic source. The bubble-trapping method is described here for asynchronous mammalian cells that grow with reasonable doubling times and from which nuclear matrices can be reliably prepared. The method for nuclear matrix preparation and enrichment of replication intermediates is described in an accompanying chapter entitled, "Purification of Restriction Fragments Containing Replication Intermediates from Mammalian Cells for 2-D Gel Analysis"). PMID- 19563115 TI - Application of alkaline sucrose gradient centrifugation in the analysis of DNA replication after DNA damage. AB - Sucrose density gradient ultracentrifugation is a powerful technique for fractionating macromolecules like DNA, RNA, and proteins. For this purpose, a sample containing a mixture of different size macromolecules is layered on the surface of a gradient whose density increases linearly from top to bottom. During centrifugation, different size macromolecules sediment through the gradient at different rates. The rate of sedimentation depends, in addition to centrifugal force, on the size, shape, and density of the macromolecules, as well as on the density and viscosity of the gradient. In this way, macromolecules are separated by size with larger ones sedimenting towards the bottom and lighter ones remaining close to the top of the gradient. The method has been particularly successful in the size fractionation of large DNA molecules and has been extensively used to measure induction and repair of DNA breaks after exposure to clastogenic factors. Here, we describe an adaptation of this method that can be used in the analysis of newly synthesized DNA formed during DNA replication. Through size analysis of nascent DNA in alkaline sucrose gradients, variations in replication activity can be measured after exposure of cells to DNA-damaging agents. The method is particularly useful as it allows distinction between DNA damage-mediated effects on chain elongation vs. replicon initiation, which is essential for an in-depth analysis of the intra-S-phase checkpoint. This ability makes the technique unique and justifies its somewhat labour-intensive nature. PMID- 19563116 TI - Isolation of recombinant DNA elongation proteins. AB - This chapter summarizes isolation procedures of four recombinant human proteins crucial for DNA replication: (a) the replicative DNA polymerase (pol) delta, (b) proliferating cell nuclear antigen (PCNA), (c) replication protein A (RP-A), and (d) replication factor C (RF-C). Pol delta is a four-subunit enzyme essential for replication of the lagging strand and possibly of the leading strand. PCNA is a central player important for coordination of the complex network of proteins interacting at the replication fork. RP-A is single-strand DNA-binding protein involved in DNA replication, DNA repair, DNA recombination, and checkpoint control. RF-C as a clamp loader is required for loading of PCNA onto double stranded DNA and therefore enables PCNA-dependent elongation by pol delta and pol epsilon. To reconstitute the intact pol delta and RF-C, a baculovirus expression system is used, where insect cells are infected with baculoviruses, each coding for one of the four or five subunits of pol delta or RF-C, respectively. We also present two easy methods to isolate the homotrimeric human PCNA and the heterotrimeric human RP-A from an Escherichia coli expression system. PMID- 19563117 TI - In vitro assays for studying helicase activities. AB - Unwinding of double-stranded DNA is required to create a single-stranded DNA template for essential DNA processes such as those involved in recombination, repair, and replication. A set of specialized enzymes called DNA helicases is dedicated to this purpose, catalyzing DNA strand separation by breaking hydrogen bonds and other noncovalent interactions that stably hold the two complementary DNA strands together. They use energy derived from the hydrolysis of nucleotide triphosphates for both bond breakage between complementary bases and translocation of a helicase enzyme along DNA. DNA unwinding activity catalyzed by a helicase usually exhibits a specific directionality (5' to 3' or 3' to 5') with respect to the DNA strand to which the enzyme is bound and moves. Unwinding activity ofa DNA helicase and its related properties can be easily measured in vitro using common lab equipment. We will describe the detailed methods and notes for preparation of various helicase substrates and in vitro helicase assays using the substrates prepared. PMID- 19563118 TI - The use of 2-aminopurine fluorescence to study DNA polymerase function. AB - The fluorescence of the base analog 2-aminopurine (2AP) is used in highly sensitive assays to determine kinetic parameters for DNA polymerase catalyzed reactions, including exonucleolytic proofreading and nucleotide binding and incorporation. Since 2AP fluorescence can also be used to probe DNA polymerase induced conformational changes in 2AP-labeled DNA substrates, reaction steps that occur before product formation can be detected. Instruction is provided here in the use of 2AP fluorescence in steady-state and presteady-state assays to study DNA polymerase function and DNA replication. PMID- 19563119 TI - Single-molecule observation of prokaryotic DNA replication. AB - Recent advances in optical imaging and molecular manipulation techniques have made it possible to observe the activity of individual enzymes and study the dynamic properties of processes that are challenging to elucidate using ensemble averaging techniques. The use of single-molecule approaches has proven to be particularly successful in the study of the dynamic interactions between the components at the replication fork. In this section, we describe the methods necessary for in vitro single-molecule studies ofprokaryotic replication systems. Through these experiments, accurate information can be obtained on the rates and processivities of DNA unwinding and polymerization. The ability to monitor in real time the progress of a single replication fork allows for the detection of short-lived, intermediate states that would be difficult to visualize in bulk phase assays. PMID- 19563120 TI - Visualization of DNA replication sites in mammalian nuclei. AB - DNA replication takes place at discrete sites in the cell nucleus, named replication foci. The spatial arrangements of these foci change in the course of S phase in a temporally regulated and reproducible fashion forming five distinct and highly conserved replication patterns. The organization of nuclear replication sites can be studied by electron and light microscopy techniques. This chapter describes several procedures for detection of replication foci in mammalian nuclei via indirect immunofluorescence microscopy. PMID- 19563121 TI - Cell-cycle synchrony for analysis of S. pombe DNA replication. AB - Analysis of S phase events requires the ability to synchronize the cell cycle for subsequent physiological or molecular analysis. The cell cycle can be arrested at different stages, using drugs or temperature-sensitive mutations, and then released (block and release). These methods offer high levels of synchrony. Synchrony is lower in methods that fractionate the cells (lactose gradient synchrony), which avoids any cell-cycle perturbation. The degree of synchronization is assessed by monitoring cell morphology or DNA content. The choice of method is influenced by the needs of the experiment and the requirements of any mutations already in the strain of interest. PMID- 19563122 TI - Measuring DNA content by flow cytometry in fission yeast. AB - Flow cytometry is an essential tool to monitor DNA content and determine cell cycle distribution. Its utility reflects the relative ease of sample preparation and the stochiometric nature of the most popular DNA-binding dyes (propidium iodide and Sytox Green). Mammalian precedents using flow cytometry for replication and cell biology studies are attractive examples for S. pombe researchers. However, the study of DNA replication with multicolor analysis has lagged behind that in mammalian cells. We present basic and advanced protocols for analysis of DNA replication in fission yeast by flow cytometry including whole cell, nuclear "ghosts," and two-color imaging with BrdU. PMID- 19563123 TI - Microscopy techniques to examine DNA replication in fission yeast. AB - Temporal and spatial visualization of replication proteins and associated structures within the narrow confines of a yeast nucleus is technically challenging. Choosing the appropriate method depends upon the parameters of the experiment, the nature of the molecules to be observed, and the hypothesis to be tested. In this chapter, we review three broad types of visualization: whole cell fluorescence or immunofluorescence, which is useful for questions of timing and chromatin association; nuclear spreads, which provide greater resolution within the chromatin for colocalization and region-specific effects; and chromatin fibers, which allow observation of labeled proteins and newly synthesized DNA on a linear chromosome. We discuss applications of these protocols and some considerations for choosing methods and fluorophores. PMID- 19563124 TI - Using the DHFR heat-inducible degron for protein inactivation in Schizosaccharomyces pombe. AB - Inactivating a specific protein in vivo can yield important information about its function. One strategy previously developed in Saccharomyces cerevisiae by the Varshavsky group involves fusing a degron, derived from mouse dihydrofolate reductase, to the N-terminus of the target protein, which thereby confers temperature-sensitive degradation at the restrictive temperature. We describe here the application of this technique in the fission yeast, Schizosaccharomyces pombe. PMID- 19563125 TI - Assays used to study the DNA replication checkpoint in fission yeast. AB - The DNA replication checkpoint, also known as the intra-S or S-phase checkpoint, plays a central role in ensuring the accuracy of DNA replication. When replication is impeded by DNA damage or other conditions, this checkpoint delays cell cycle progression and coordinates resumption of replication with DNA repair pathways. One of its critical functions is to stabilize stalled replication forks in a replication-competent state, presumably by maintaining proper assembly of replisome components and preserving DNA structures. Here we describe a series of assays used to study the replication checkpoint. These assays allow us to investigate the specific functions of proteins involved in the replication checkpoint in fission yeast. PMID- 19563126 TI - Incorporation of thymidine analogs for studying replication kinetics in fission yeast. AB - Labeling DNA during in vivo replication by the incorporation of exogenous thymidine and thymidine analogs has been a mainstay of DNA replication and repair studies for decades. Unfortunately, thymidine labeling does not work in fungi, because they lack the thymidine salvage pathway required for uptake of exogenous thymidine. This obstacle to thymidine labeling has been overcome in yeast by engineering a minimal thymidine salvage pathway consisting of a nucleoside transporter to allow uptake of exogenous thymidine from the medium and a thymidine kinase to phosphorylate the thymidine into thymidine monophosphate, which can be used by the cell. This chapter describes the labeling of fission yeast, Schizosaccharomyces pombe, with the thymidine analog BrdU in order to identify sites and determine kinetics of DNA replication. PMID- 19563127 TI - The fast-halo assay for the assessment of DNA damage at the single-cell level. AB - The detection of breaks in mammalian cell DNA and the measurement of their repair represent primary endpoints for genotoxicity testing. Over the past three decades many techniques sensitive to the presence of DNA breaks have been developed: their availability significantly increased the knowledge in the area of genetic toxicology. In general, these techniques have evolved to become more sensitive and flexible as well as less complicated. The fast-halo assay (FHA) is a very recent method to detect DNA-strand breakage induced either by various genotoxic agents or secondary to apoptotic DNA cleavage, and to study the repair of primary DNA breaks at the single-cell level. In FHA, damaged DNA is separated from intact one by means of solvent gradient, stained with ethidium bromide and visualized under a fluorescence microscope. The level of DNA breaks is then determined with an image analysis software. FHA is as sensitive, reliable, and flexible as the well-established comet assay, but it has the advantage of being, as compared to any other existing method, the most rapid and less expensive one. Taken collectively, these unique features render FHA the ideal method to perform a large number of genotoxicity tests on mammalian cells in a particularly cost effective and time-saving manner. PMID- 19563128 TI - Monitoring homologous recombination following replication fork perturbation in the fission yeast Schizosaccharomyces pombe. AB - Replication forks (RFs) frequently encounter barriers or lesions in template DNA that can cause them to stall and/or break. Efficient genome duplication therefore depends on multiple mechanisms that variously act to stabilize, repair, and restart perturbed RFs. Integral to at least some of these mechanisms are homologous recombination (HR) proteins, but our knowledge of how they act to ensure high-fidelity genome replication remains incomplete. To help better understand the relationship between DNA replication and HR, fission yeast strains have been engineered to contain intrachromosmal recombination substrates consisting of non-tandem direct repeats of ade6 heteroalleles. The substrates have been modified to include site-specific RF barriers within the duplication. Importantly, direct repeat recombinants appear to arise predominantly during DNA replication via sister chromatid interactions and are induced by factors that perturb RFs. Using simple plating experiments to assay recombinant formation, these strains have proved to be useful tools in monitoring the effects of impeding RFs on HR and its genetic control. The strains are available on request, and here we describe in detail how some of them can be used to determine the effect of your mutation of choice on spontaneous, DNA damage-induced, and replication block-induced recombinant formation. PMID- 19563129 TI - Computational methods to study kinetics of DNA replication. AB - New technologies such as DNA combing have led to the availability of large quantities of data that describe the state of DNA while undergoing replication in S phase. In this chapter, we describe methods used to extract various parameters of replication--fork velocity, origin initiation rate, fork density, numbers of potential and utilized origins--from such data. We first present a version of the technique that applies to "ideal" data. We then show how to deal with, a number of real-world complications, such as the asynchrony of starting times of a population of cells, the finite length of fragments used in the analysis, and the finite amount of DNA in a chromosome. PMID- 19563130 TI - Use of DNA combing to study DNA replication in Xenopus and human cell-free systems. AB - The Xenopus egg extract has become the gold standard for in vitro studies of metazoan DNA replication. We have used this system to study the mechanisms that ensure rapid and complete DNA replication despite random initiation during Xenopus early development. To this end we adapted the DNA combing technique to investigate the distribution of replication bubbles along single DNA molecules. DNA replicating in egg extracts is labelled by addition of digoxigenin-11-dUTP and/or biotin-16-dUTP at precise times. These two dTTP analogues are efficiently incorporated into DNA during replication in the extract. After DNA purification and combing the DNA is visualized with appropriate fluorescent antibody/streptavidin molecules. Replicated DNA appears as green or red tracts whose pattern reveals how each molecule was replicated, allowing to follow the dynamics of DNA replication through S phase. We describe (a) the preparation and use of egg extracts and demembranated sperm chromatin templates; (b) a simple method for preparing silanized glass coverslips suitable for DNA combing and fluorescence detection; (c) two alternative replicative DNA labelling schemes and their respective advantages; and (d) a protocol for combining replicative labelling with detection of specific DNA sequences by fluorescent in situ hybridization (FISH). Although most observations made in Xenopus egg extracts are applicable to other eukaryotes, there are differences in cell-cycle regulation between mammalian somatic cells and embryonic amphibian cells, which led to the development of human cell-free systems that can initiate semi-conservative chromosomal DNA replication under cell-cycle control. We have employed the knowledge gained with Xenopus extracts to characterize DNA replication intermediates generated in human cell-free systems using DNA combing. We describe here (a) the preparation and use of human cell-free extracts and initiation competent template nuclei for DNA combing studies; (b) an optimized labelling scheme for DNA replication intermediates by molecular combing and fluorescence microscopy. PMID- 19563131 TI - Electron microscopy methods for studying in vivo DNA replication intermediates. AB - The detailed understanding of the DNA replication process requires structural insight. The combination of psoralen crosslinking and electron microscopy has been extensively exploited to reveal the fine architecture of in vivo DNA replication intermediates. This approach proved instrumental to uncover the basic mechanisms of DNA duplication, as well as the perturbation of this process by genotoxic treatments. The replication structures need to the stabilized in vivo (by psoralen crosslinking) prior to extraction and enrichment procedures, finally leading to the visualization at the transmission electron microscope. This chapter outlines the procedures required to visualize in vivo replication intermediates of genomic DNA, extracted from budding yeast or cultured mammalian cells. PMID- 19563132 TI - Determining the replication dynamics of specific gene loci by single-molecule analysis of replicated DNA. AB - In metazoans, development and cell differentiation are known to affect various aspects of chromosomal organization at developmentally regulated gene loci (e.g., nuclear localization, locus accessibility, chromatin modifications, etc.). Recent evidence also indicates that cell differentiation can have dramatic effects on DNA replication initiation and replication fork progression, at several of these loci. Hence, origin selection and activation are no longer viewed as constitutive events, but as actively regulated processes, the deregulation of which could potentially affect gene expression and genomic stability at particular chromosomal locations. For these reasons, the study of DNA replication across large genomic regions has recently received increasing attention. Here we describe an assay based on the analysis of single DNA molecules that can be used to determine the steady-state distribution of replication forks across large genomic regions. This assay can be used to study how DNA replication initiates, progresses, pauses, and terminates at single-copy loci in mammalian cells. PMID- 19563134 TI - The need for a national health policy, plan and performance monitoring in health development. PMID- 19563133 TI - Use of DNA combing for studying DNA replication in vivo in yeast and mammalian cells. AB - Plasticity is an inherent feature of chromosomal DNA replication in eukaryotes. Potential origins of DNA replication are made in excess, but are used (fired) in a partly stochastic, partly programmed manner throughout the S phase of the cell cycle. Since most origins have a firing efficiency below 50%, population-based analysis methods yield a cumulative picture of origin activity (obtained by accretion) that does not accurately describe how chromosomes are replicated in single cells. DNA combing is a method that allows the alignment on silanized glass coverslips, at high density and with uniform stretching, of single DNA molecules in the Mb range. If this DNA is isolated from cells that have been labelled with halogenated nucleotides (BrdU, CldU, IdU), it is possible to determine the density and position of replication origins as well as the rate and symmetry of fork progression, quantitatively and on single DNA molecules. This chapter will successively describe (a) the preparation ofsilanized coverslips, (b) the incorporation of halogenated nucleotides in newly synthesized DNA in yeast and mammalian cell lines, (c) the preparation and combing of genomic DNA, and finally (d) the acquisition and analysis of single-molecule images to extract salient features of replication dynamics. PMID- 19563135 TI - A review of national health policies in seven countries in the WHO African Region. AB - OBJECTIVES: This paper proposes an analytical framework for assessing compliance of national health policies with WHO/AFRO guidelines. DATA SOURCES: Data for this study was obtained from the national health policies of Botswana, Eritrea, Liberia, Namibia, Swaziland, Gambia, and Uganda. STUDY SELECTIONS: National health policies of seven of the 19 Anglophone countries of the WHO African region were selected for review using simple random sampling method. These include: Botswana, Eritrea, Liberia, Namibia, Swaziland, Gambia, and Uganda. DATA EXTRACTION: An analytical framework derived from WHO/AFRO guidelines for developing national health policies and plans was used in the review. It identifies components which are pertinent for appropriate national health policy formulation. DATA SYNTHESIS: It appears that aspects related to policy content are well addressed. In relation to the process, there is need for improving the mapping of stakeholders and specifying their roles and aspects of collaboration; and the implications for meeting broad service and impact targets. CONCLUSION: Development of health policies needs to focus on all aspects of the analytical framework with emphasis on improving the articulation for mapping out stakeholders and specifying their roles and aspects of collaboration; and the implications for meeting broad service and impact targets. PMID- 19563136 TI - Status of health sector strategic plans in five countries of the WHO African Region. AB - OBJECTIVES: To assess the adequacy of the existing strategic plans and compare the format and content of health sector strategic plans with the guidelines in selected countries of the African region. DATA SOURCE: The health strategic plans for Gambia, Liberia, Malawi, Tanzania and Uganda, which are kept at the WHO/AFRO, were reviewed. DATA EXTRACTION: All health strategic plans among the Anglophone countries (Gambia, Ghana, Kenya, Liberia, Malawi, Mauritius, Tanzania, Uganda, Zambia and Zimbabwe) that were developed after the year 2000 were eligible for inclusion. Fifty percent of these countries that fitted this criterion were randomly selected. They included Gambia, Liberia, Malawi, Tanzania and Uganda. The analysis framework used in the review included situation analysis; an assessment of appropriateness of strategies that are selected; well developed indicators for each strategy; the match between the service and outcomes targets with available resources; and existence of a clear framework for partnership engagement for implementation. DATA SYNTHESIS: Most of the strategic plans identify key ill health conditions and their contributing factors. Health service and resource gaps are described but not quantified in the Botswana, Gambia, Malawi, Tanzania strategic documents. Most of the plans selected strategies that related to the situational analysis. Generally, countries' plans had clear indicators. Matching service and outcome targets to available resources was the least addressed area in majority of the plans. Most of the strategic plans identified stakeholders and acknowledged their participation in the implementation, providing different levels of comprehensiveness. CONCLUSION: Some of the areas that are well addressed according to the analysis framework included: addressing the strategic concerns of the health policies; identifying key partners for implementation; and selection of appropriate strategies. The following areas needed more emphasis: quantification of health system gaps; setting targets that are cognisant of the local resource base; and being more explicit in what stakeholders' roles are during the implementation period. PMID- 19563137 TI - A rapid assessment of district health systems in six countries of the WHO African Region. AB - OBJECTIVES: This paper reviews the adequacy of inputs and processes at district level to support outputs and outcomes of service delivery at district level using a rapid assessment. The outputs included in this study are those considered essential for the attainment of the Health related Millennium Development Goals (MDGs). DATA SOURCES: A questionnaire based rapid District Health Systems assessment was conducted among six African countries during the year 2007. STUDY SELECTIONS: The study took place in a random sample of six out of 19 English speaking countries of the WHO African region. These countries are Ghana, Liberia, Namibia, Nigeria, Sierra Leone and Uganda. DATA EXTRACTION: The data was extracted from the questionnaires, entered and analysed in Excel spreadsheet. DATA SYNTHESIS: In spite of the variability in quality and completeness of reporting on the selected parameters, this paper does indicate that according to country norms and standards, the inputs and processes are insufficient to lead to acceptable outputs and outcomes, especially those related to the MDGs. An important point to note is that comparability across countries is made on the basis of individual country norms and standards. Implicit in this assessment is that country norms and standards are reasonable and are appropriate for the attainment of the MDGs. However reasonable the country norms and standard are, it is unlikely that the low resource base as well as weak organisational and managerial capacities in most countries will support effectively the attainment of the MDGs. CONCLUSION: Most countries manage to offer the essential health services at all levels of care despite the relatively low level of inputs. However, their level of quality and equity is debatable. The general trend is that provision of the essential health services is more at the higher levels of care prompting concerns for the populations served at lower levels of care. There is also a tendency to have wide variations in the performance of service delivery geographically as well as at the different levels of the health systems. This paper recommends further exploration of the impact of focusing on improving quality of existing health services while increasing quantity of service delivery points to achieve higher coverage of essential health services. PMID- 19563138 TI - Monitoring and evaluation of health sector reforms in the WHO African Region. AB - OBJECTIVES: To describe a comprehensive analytical framework for assessing health sector reforms and demonstrates use of the analysis framework using cost-recovery mechanisms as a case study in the WHO African Region. DATA SOURCES: Health sector reforms published literature review. STUDY SELECTION: No selection involved. DATA EXTRACTION: This paper draws from previous published literature to describe a comprehensive framework to assess the performance of health sector reforms in the African region. Using this framework, it goes on to illustrate how it may be used to analyze cost recovery reforms as a case study. The major elements for the analysis include a description of the context, design, process and intended results. DATA SYNTHESIS: In terms of context and design of the cost recovery reform, there were gaps in the stewardship role of governments as evidenced by the lack of appropriate policies and information to monitor and/or influence the process. Regarding the cost recovery implementation, it is not clear from the literature reviewed in this paper that there was a comprehensive stakeholder coordination mechanism that catered for all who were involved. Concerning results of the expected results of implementing cost recovery reforms such as improved quality of health services; equitable service utilisation; social sustainability through active community participation; and gains in efficiency were not always realised. CONCLUSIONS: Given that the aspects of the analysis framework described in this paper are interrelated, reviewing one without another provides an answer to a specific question but is insufficient for a comprehensive assessment. PMID- 19563139 TI - Croatian Adult Health Survey--a tool for periodic cardiovascular risk factors surveillance. AB - National risk factors surveys present very important tool for gathering population based health related information for policy. Croatian Adult Health Survey (CAHS) is such a tool, intended to collect population-based, representative and authoritative information on cardiovascular risk factors prevalence. The CAHS sample was designed to provide nearly complete coverage (98%) of the Croatian adult population, based on the random selection of the adult member of the household that was selected in the complex sampling scheme. Additionally, seven-step weighting scheme was applied to the dataset, in order to further increase the representativeness of the sample which is consequently considered to be representative for six geographical and administrative regions of Croatia, while it is considered only to be informative for the county-based estimates. The first cycle of the project took place in 2003, with a total of 9,070 respondents. The second project cycle is taking place in 2008. It has now been converted into the follow-up study (re-surveying the 2003 sample) and is being further supplemented with additional questions, qualitative investigation module and an intervention performed by the public health nurses. The CAHS provides not only the basis for health information for policy, but also serves as a platform for a number of ongoing public health collaborations and a valuable public health research resource. PMID- 19563140 TI - Regional pattern of cardiovascular risk burden in Croatia. AB - The objective of this study was to investigate regional gradient of a synthetic cardiovascular burden (CVRB) in Croatia. Analysis was based on a multistage stratified sample representative of general adult population in six regions of Croatia (Croatian Adult Health Survey). Synthetic CVRB was defined by incidents (heart attack, stroke), blood pressure, overweight/obesity (BMI, waist circumference), and risky behaviours (smoking, physical inactivity, high alcohol consumption, inadequate nutrition). Total CVRB in Croatia was 44.7% for men and 50.3% for women. Combining both high and moderate CVRB, men were found to be under more risk than women (72.8% vs. 61.7%, respectively). The result showed an east-north-west gradient in continental Croatia, with high CVRB in Eastern (40.2%) and Central region (44.3%), City of Zagreb (45.7%) and Northern region (53.1%). Mountainous region had the lowest prevalence of respondents with high CVRB (39.2%). Coastal region with the prevalence of 46.3% of high CVRB was nearly the same as the City of Zagreb. The results suggest the presence of substantial regional differences in the cardiovascular risk burden. PMID- 19563141 TI - Regional differences in the prevalence of arterial hypertension in Croatia. AB - This paper analyzes the Croatian Adult Health Survey data, collected in 2003 with a total of 9,070 respondents aged 18+. Based on an average of two measurements, respondents with the mean systolic arterial pressure > or = 140 mmHg or mean diastolic pressure > or = 90 mmHg were classified as hypertensive. The data for men and women were analyzed separately, according to regions. Prevalence of hypertension in men was 40.5% (95% confidence interval CI 37.9-43.01; coefficient of variability CV = 3.2), women 34.9% (95% CI 33.2-36.7; CV 2.5). There were no significant differences in regional prevalence in men, except in the Northern and Eastern region. In women we did not detect any significant regional difference. Non-controlled arterial hypertension is an important public health problem in all monitored regions of Croatia. Raising awareness about the problem, early detection and encouraging the population to adhere to the therapy for elevated arterial pressure, in addition to a healthy life style, are important for successful control and harm reduction. PMID- 19563142 TI - The prevalence of overweight, obesity and central obesity in six regions of Croatia: results from the Croatian Adult Health Survey. AB - Our aim was to estimate the prevalence of overweight, obesity and increased waist circumference in the adult population of Croatia and investigate regional differences in six regions of Croatia. Using the data from the 2003 Croatian Adult Health Survey we estimated the overall prevalence of overweight, obesity and increased waist circumference for the entire population of Croatia at 38.11%, 20.34% and 43.52%, respectively. For men, this was 43.2%, 20.1% and 34.98% and for women 33.6%, 20.6% and 51.13%, respectively for the three indicators of increased body weight. We found conflicting evidence as to whether the Mediterranean part of the country, compared with the continental part, bears a lesser degree of cardiovascular risk. Planners should pay particular attention to the Northern region, where the burden of increased body weight was the highest. PMID- 19563144 TI - Regional pattern of physical inactivity in Croatia. AB - The aim of this paper was to analyze the regional pattern of physical inactivity in Croatia based on the Croatian Adult Health Survey 2003 data. A total of 9,070 adult respondents were included in this study. In men, the highest prevalence of physical inactivity was recorded in the City of Zagreb (39.6%), and it was significantly higher than in Central (25.6%), Coastal (25.6%) and Mountainous region (14.1%). Mountainous region had significantly lower prevalence of physical inactivity compared to any other region, except the Central region. The highest prevalence of physical inactivity in women was also recorded in the City of Zagreb (43.6%), and it was also significantly higher than in all other regions. The lowest prevalence of physical inactivity was recorded in Eastern Region (24.7%). The highest levels of physical inactivity in both in both genders were recorded in urban regions, suggesting that intervention measures in terms of health promotion should be undertaken, with strong emphasis on the people living in urban settings. PMID- 19563143 TI - Regional differences in dietary habits of adult Croatian population. AB - The aim of this study was to investigate the dietary habits of the adult Croatian population according to geographical regions and gender. Date was obtained from the Croatian Adult Health Survey questionnaire, carried out in six regions of Republic of Croatia (Eastern, Northern, Central, City of Zagreb, Mountainous and Coastal), with a total of 9,070 responses. Results have shown that the highest prevalence of unhealthy dietary habits was recorded in Eastern and Central regions, while Coastal region and city of Zagreb had significantly lower prevalence of unhealthy dietary habits (P < 0.05). Prevalence of unhealthy dietary habits was higher in men in all regions. Women had healthier dietary habits, what was strongly expressed in Coastal and Mountainous region. This study has shown that the unhealthy dietary habits were practiced by approximately one quarter of the adult population, regardless on the region in which they resided. Regional differences in dietary habits should be taken into account in any new public health studies and interventions. PMID- 19563145 TI - Regional differences in alcohol consumption in Croatia. AB - This study aims to estimate regional and gender differences in the prevalence of alcohol consumption in Croatia. It is based on the data obtained from the Croatian Adult Health Survey 2003. The highest prevalence of alcohol consumption in men was reported in Eastern Croatia (14.1%). Men living in the Mountainous region had the lowest prevalence of reported alcohol consumption (8.8%). In contrast to men in the Eastern region, women in the same region reported drinking alcoholic beverages with the lowest prevalence (0.3%). The highest proportion of women who reported drinking alcoholic beverages was recorded the Northern Croatian region (1.5%). Results support the expected gender gap in alcohol consumption and point out strong regional pattern of alcohol consumption. The observed pattern shows that primary prevention and health promotion should be implemented with special concern to regional differences and traditions. PMID- 19563146 TI - Regional pattern of smoking in Croatia. AB - The aim of this paper was to investigate the regional smoking pattern in Croatia. The data from the Croatian Adult Health Survey from 2003 were used for the study (N = 9,070). The results of this study suggest that the prevalence of smoking among men was the highest in the Eastern parts of the country, followed by the Mountainous region. In contrast, the prevalence of smoking among women was the highest in City of Zagreb. The results support the strong regional pattern, with more pronounced differences in women (those from the City of Zagreb had more than twice higher prevalence than those from the Northern parts of the country). Smoking is still relatively highly prevalent in Croatian adult population (24-33% in men, and between 10 and 21% in women, depend on region). This suggesting that preventive actions and public health interventions are needed to reduce the smoking prevalence and thus improve the population health. PMID- 19563147 TI - Trends and regional disparities in ischemic heart disease and cerebrovascular disease in Croatia, 1997-2006. AB - The aim of this study was to analyze mortality and hospital morbidity from ischemic heart disease (IHD) and cerebrovascular disease (CVD) in the population aged 18+ years in different regions of Croatia, in 1997 to 2006 period. Mortality data were supplied by Central Bureau of Statistics, while hospital morbidity data were obtained from hospital discharge database at Croatian National Institute of Public Health. In Mediterranean region rates of IHD mortality were lower, while rates of hospitalization due to IHD have been higher than those for Croatia in the entire period. Rates of IHD mortality for Continental part have not been different from mean rates for Croatia while rates of hospitalization due to IHD have been lower than rates for Croatia in entire period. Rates of CVD mortality and rates of hospitalization due to CVD for Mediterranean part have been lower than rates for Croatia, while both rates for Continental part have been higher than rates for Croatia during entire period. This analysis identified the regions in which significant deviations from the general trend suggest the need for creation of a new national public health intervention model, focusing on changeable behaviour (lifestyles) risk factors. PMID- 19563148 TI - Characteristics of heavy smokers in Croatia. AB - The aim of this paper was to investigate the socio-demographic characteristics of heavy smokers in comparison to group the characteristics of light smokers and non smokers. Data used for this study was collected within the Croatian Adult Health Survey (CAHS). The results show a significant gender differences in the prevalence of heavy smoking, with higher prevalence among men in all age groups. Highest prevalence of heavy smoking was found in people 35 to 64 years of age, irrespective to gender. According to analyzed socio-demographic characteristics men with low education and income who changed place of living have higher prevalence of heavy smoking. Pattern was different in women smokers. Majority of women smokers were light smokers. In women with better than average household economic status there were more smokers than non-smokers, with notable high proportion of heavy smokers in age 18 to 34 years and light smokers in age group 35 to 64. While planning future interventions at the population level, we must consider all of these characteristics and links between them, and engage all segments of society. PMID- 19563149 TI - Health behavior factors associated with obesity in the adult population in Croatia. AB - The aim of this study was to examine the association of obesity and selected health behaviors, based on the data from Croatian Adult Health Survey 2003. Cross sectional study on representative random sample of 9070 Croatian adults showed that in both men and women, mean BMI varied significantly with age (p < 0.001). It tended to increased with age until 55-64 years, and then decreased slightly in men, but remained unchanged in women. The prevalence of obesity (BMI > or = 30.0 kg m(-2)) demonstrated almost the same prevalence in men and women, 20.1% and 20.6% respectively. The likelihood of being obese, either in men or women, was more likely in the middle-aged groups, among respondents from rural areas and those from the Continental region of Croatia, with drinking lifestyle pattern, and non-smokers. Women who mostly used animal fat for food preparations were more likely to be obese. Overweight and obesity are major public health problem in the adult population in Croatia, and health promotion strategies based on behavioral correlates are needed to prevent excess weight gain in the Croatian population. PMID- 19563150 TI - Anthropometric indices of obesity and hypertension in different age and gender groups of Croatian population. AB - The aim of this study was to investigate the association of hypertension with the obesity indices--body mass index and waist circumference. The study was based on the data from the Croatian Adult Health Survey 2003, which is a cross-sectional study on the representative sample of 9,070 Croatian adults. The results showed that in both men and women aged 18-64 years and 65 years and older increased BMI and waist circumference were associated with higher odds of hypertension. Odds of hypertension for obese (BMI > or = 30) and persons with abdominal obesity (waist circumference > or = 102 cm for men, > or = 88 cm for women) was higher among young and middle aged than among elderly persons. No differences in odds of hypertension were recorded between persons with increased BMI and persons with increased waist circumference. Both BMI and waist circumference exhibited equally strong association with hypertension in Croatian population, in both men and women, and in those who are below or over 65 years of age. PMID- 19563151 TI - Primary and secondary prevention of cardiovascular diseases embedded in the visiting nurse services: description of the intervention model. AB - The paper describes a visiting nurse led intervention model for the primary and secondary prevention of cardiovascular diseases (CVD) and specificities of its application. Although CVD burden is high in Croatia, the visiting nurse services have not been specifically focused on CVD prevention in the population until now. The intervention model described here is being implemented alongside the second cycle of the Croatian Adult Health Survey (2008 CAHS). The model includes an objective evaluation of respondents' CVD risk factors through quantitative and qualitative analyses, as well as respondents' self-evaluation of risk factors and motivation to change. At the same time, respondents are educated and intervention is evaluated. A 'health booklet' was specifically designed for documentation during one year's follow-up, where both the user and the visiting nurse keep copies of the negotiated targets and strategies set to achieve them. This intervention model has the potential to mobilize the service towards permanent incorporation of primary and secondary CVD prevention into routine care and, due to work specificities of the visiting nurse services, to cover the entire population in an organized CVD prevention. PMID- 19563152 TI - Cardiovascular diseases, risk factors and barriers in their prevention in Croatia. AB - Cardiovascular diseases are the leading cause of death in Croatia, with significant regional differences. Despite high mortality rates, high prevalence of various cardiovascular risk factors and well organized public health network, comprehensive system for cardiovascular disease monitoring and interventions does not exist. In this study we analyzed legislation framework and responsibilities of stakeholders relevant for cardiovascular disease surveillance and prevention. According to the international experiences we analyzed characteristics of cardiovascular disease prevention in Croatia and causes of the problems appeared in the preventive programs in Croatia. Analysis showed that primary problem is not inefficiency, but the existence of barriers in preventive activities definition, responsibilities distribution and task implementation. Main cause for such situation is incompatibility of the existing practices in clinical medicine and public health with recommendations from other countries. For the successful prevention of cardiovascular disease in Croatia at least three changes need to be made--define new terms and contents of prevention, define new responsibilities distribution and provide equity in health as basic criterion for successful preventive programs. PMID- 19563153 TI - Psychological distress within cardiovascular risks behaviors, conditions and diseases conceptual framework. AB - Psychological distress (PD) is being increasingly recognized as a risk factor for cardiovascular diseases (CVD). Our aim was to recognize an association of PD and CVD in the Croatian adult population. We also explored association's strength obtainable as relative risk of PD on three levels; cardiovascular risk behaviors, conditions and diseases. This study used Croatian Adult Health Survey 2003 (CAHS 2003) data (N = 9,070). PD status was measured by the five-item Mental Health Scale of the Short Form questionnaire (SF-36) hence one distinguished subgroup consisted of population with PD and other without PD. Prevalence of cardiovascular risk behaviors, cardiovascular risk conditions and self-reported cardiovascular diseases within each subgroup were calculated using bootstrap method. Women had higher prevalence of PD in general population. Among distressed population women had higher prevalence of body mass index over 30, metabolic syndrome and angina pectoris. Men with PD had higher prevalence of high blood pressure and myocardial infarction with contradictory lower prevalence of angina pectoris then myocardial infarction. Physical inactivity was proven to be a risk behavior determinant with most impact on mental health. All CVD are consistently associated with higher prevalence and relative risks for PD both in men and women. PMID- 19563154 TI - Cardiovascular and behavioral risk factors in relation to self-assessed health status. AB - The purpose of this study was to explore biomedical and behavioral risk factors in relation to self-assessed physical, mental and general health status in an open adult Croatian population sample. Subjective experience of health status was assessed with the Short Form 36 Health Survey questionnaire (SF-36). Out of 9070 respondents, we defined two groups with respect to cardiovascular (CV) risk factors: (1) a healthy group of individuals who did not state the presence of any covered chronic disease or disorder (N = 1,817), and (2) a group with CV risk which included individuals who reported having high blood pressure or high blood cholesterol or high blood sugar diagnosed (N = 360). When adjusted for socio demographic characteristics, these two groups differed in self-assessed health status. The group with CV risk factors showed an average lower level of subjective health status than the healthy group. At the level of specific health dimensions, the group with CV risks reported significantly lower general and mental health, but they reported healthier behaviors at the present time. We analyzed the measured health behaviors in predicting individual differences in the physical, mental and general health of the healthy group. Physical activity was revealed as a significant predictor of all three aspects of subjective health. Socio-economic variables of age, gender and self-assessed economic status contributed significantly to the explanation of all three aspects of subjective health. Our findings emphasize that psychological, physical, and social factors are inextricably linked in maintaining cardiovascular health, thus showing the importance of targeting health-related behaviors, especially physical activity, in preventive strategies and programs. PMID- 19563155 TI - Health related quality of life of smokers in Croatia. AB - The aim of this study was to investigate the health related quality of life (HRQOL) in relation to the smoking status. The data from the Croatian Adult Health Survey conducted in 2003 were used for this study. Sample comprised 9,070 participants, 68.1% women and 31.9% men, from 18 to 101 years old. Results indicated that there were significant differences in HRQOL between smokers and non-smokers even in a young adult age when there is no difference in objective health status (for example diagnosis of disease). Generally health decreased with the age for both groups but direction of difference between smokers and non smokers varied for different health dimensions across the age groups. The most notable difference in HRQOL in relation to smoking was found in the age group of 65 and older where women smokers reported better HRQOL than non-smokers, and men smokers significantly worse HRQOL than non-smokers. As we found significant differences according to gender in age groups, we suggest that it is essential for future studies to take those characteristics into consideration. Many other factors, for example education, income, marital status, and the socio-cultural acceptability of smoking may have an impact on the quality of life, and also need to be considered in future research. The existing differences in mental health and social functioning should be taken into account in planning of the future prevention programs. PMID- 19563156 TI - Lifestyle habits of Croatian diabetic population: observations from the Croatian Adult Health Survey. AB - The aim of this study was to assess the behavioural risk factors in Croatian diabetic population and to compare them with the lifestyle habits of individuals with no known history of diabetes. The study was a part of the Croatian Adult Health Survey (CAHS), a cross-sectional survey that provided comprehensive health assessment of the Croatian adult population. Risk factors were defined as an unhealthy nutritional regimen, excessive alcohol consumption, smoking and lack of physical activity. Physical inactivity was the most prevalent risk factor observed in a significant number of both diabetic and non-diabetic subjects (44.8% and 29.1%). It was also the only behavioural risk factor that was more prevalent in the diabetic individuals as compared to those without diabetes. Alcohol consumption did not vary significantly between the two groups (5.8% vs. 6.3%), while unhealthy dietary pattern and smoking were less frequent in respondents with diabetes (10.0% vs. 16.5% and 14.3% vs. 23.2%, respectively). Among diabetic patients, a significantly larger proportion of men than women reported smoking (19.2% vs. 10.0%), whereas no such sex-related differences were observed in other behavioural risk factors. Although the most prominent risk factor in diabetic patients was physical inactivity, a significant proportion of respondents with diabetes also reported the presence of other risk factors investigated in this survey. Since the majority of diabetic patients do not reach their treatment goals, there is a substantial need for curative and preventive interventions. Given the importance of physical activity in the treatment and prevention of diabetes and the high proportion of inactive diabetic patients, any future preventive programme in Croatia should address that risk as well. PMID- 19563157 TI - Health care needs, utilization and barriers in croatia--regional and urban-rural differences. AB - Even the most socially aware countries in the World have noticed the gap increase between the poorest and the richest population groups. The purpose of this study was to investigate the presence of inequity and to identify main barriers for equitable health care utilization by economic status, region and area of living, controlled for health needs in the Croatian adult population. The data from the Croatian Adult Health Survey 2003 were used in this study. The results show that among the respondents with higher health needs, those with economic status above average had higher proportion of regular annual general practitioner and medical specialist visits. In contrast, highly frequent visits to physician were more common in respondents who were below average economic status. Economically worse off women, regardless on their health care needs reported gynecologist visits less regularly than the better-off women. Long waiting and the large distance from the health care facilities were the most commonly reported barriers in health care utilization. High expenses were present as the main barrier at dentist and inpatient health services utilization. Suburban and rural settlements were more burdened with long distance from the health care facilities and high expenses for all health services, aggravated by the long waiting time for visits to GP. Respondents from the urban settings reported long waiting time and unkindness of the health personnel as the main barriers. The results of this study clearly show the main barriers in the equitable health care delivery to Croatia population from the health care users perspective. PMID- 19563158 TI - Co-occurrence of risky lifestyle behavior with overweight, excess abdominal fat and high blood pressure--case oriented approach. AB - The objective was to estimate the proportion of cases developed interim risk factors (INTF: overweight, excess abdominal fat, high blood pressure) in relation with behavioral risk factors (BEHF: smoking, heavy alcohol intake, unhealthy diet, physical inactivity). NOBIR group was defined as cases with no BEHF and BIR as those with them. Both groups show higher proportions of INTF in older age. The increase by age varies of twofold (overweight: 13.2-29.2 for men, 18.1-42.6 for women) to six fold (high blood pressure: 4.6-26.5 for men, 6.6-40.8 for women) in proportions. Women show higher proportions of INTF than men in both groups, but BIR group shows higher proportions than NOBIR in all the age groups taking the both gender together. As a BEHF the physical inactivity has a markedly increase with age (from 4% to more than 25%). Smoking is the only BEHF decreasing in oldest for all the INTF. PMID- 19563159 TI - Prevalence of increased body weight and hypertension in the population of Croatian mainland and Adriatic Islands--are islanders really healthier? AB - The aim of this study was to compare the prevalence of the hypertension and the increased body weight (BMI) between Croatian mainland and Adriatic island population. The data from the Croatian Adult Health Survey (N = 9,070) served as an estimate for the mainland Croatian population, while the data from "1001 Dalmatian study" (N = 1,001) were collected from four Adriatic islands; Rab, Vis, Lastovo and Mljet. The prevalence of increased body weight and hypertension was calculated for the four age groups and analyzed using chi-square test. The results indicate that men from the islands less frequently had normal body mass index (P < 0.001), and were more frequently overweight (P < 0.001). The prevalence of overweight and obesity were similar between the island and mainland women. The percent of normotensive respondents in men was significantly lower in islands (P < 0.001), while the prevalence of newly diagnosed hypertension was significantly higher among islanders in both genders (P < 0.001). Despite the traditionally prevalent Mediterranean diet and overall more favorable lifestyle islanders may not be as healthy as previously studies suggested, in terms of cardiovascular risk factors prevalence. This might be related to the poor access to health care and preventive measures or low interest for health care especially among men on the islands, reflected in the higher prevalence of newly diagnosed hypertension. These findings suggest that island populations represent good candidates for disease awareness programs and health promotion interventions. PMID- 19563160 TI - Healthy migrant effect within Croatia. AB - The aim of this study was to investigate the demographic determinants and health status of within-country migrants in Croatia. Data from the Croatian Adult Health Survey from 2003 (N = 9,070) were used in the analysis. Migrants due to 1991-1995 war were excluded from the analysis and only those who indicated that the reason for their migration within Croatia was related to employment or other reasons were included. Prevalence of hypertension and increased body weight was calculated for each of the three migration-related groups (employment-related migrants, other reasons migrants and non-migrants). Migrants were on general younger than non-migrants (more than three quarters of migrant were under 50 years). Employment-related migrants had the lowest prevalence of measured hypertension, while other reasons migrants had the lowest prevalence of increased body weight. Non-migrants had the worse indices in almost every trait and age group that was analysed. The results support the healthy migrant hypothesis, even in case of migrants who migrated within Croatia. However, the results of this study show that health status of migrants may depend on the reasons for their migration, suggesting that future migration studies should record and take this into account. PMID- 19563162 TI - Do public health surveys provide representative data? Comparison of three different sampling approaches in the adult population of Croatia. AB - We investigated the sample representativeness in three different types of population-based public health surveys in Croatia. Responses from the household sampling based Croatian Adult Health Survey (CAHS), health insurance register based Croatian Health Survey (CHS) and a telephone survey (TPS) were analysed and compared to gender, age and education composition of the Croatian adult population, based on the 2001 Census. The raw (unweighted) survey data were used and analysed with Spearman's rank test and distance analysis. The results indicated that TPS had the most similar gender composition compared to the Census data. TPS also had the most similar age composition in men, while CHS had the most similar age composition in women. Finally, CAHS had the most similar education composition to the Census data. Three population subgroups were substantially under-sampled in all three surveys--men, younger people, and elderly from the lowest educational classes. For these sub-groups, advanced sampling methods should be employed in order to obtain more precise estimates from public health surveys. PMID- 19563161 TI - The geographic patterns of the exceptional longevity in Croatia. AB - The aim of this study was to provide an overview of the exceptional longevity patterns in Croatia. The prevalence of nonagenarians (aged 90 years or more) and centenarians (100 years or more) was calculated from the 1953-2001 census data. The data were analyzed with chi-square test and trend analysis. The results indicate steady and significant increasing trends in both age groups, more pronounced in women. Geographical distribution of the exceptional longevity had a clear coastline-to-mainland gradient, with higher prevalence of exceptionally old people in the coastal parts of the country. Additionally, exceptionally old respondents from the Croatian Adult Health Survey (CAHS) were compared to elderly ones (65-89 years old). The results suggested that exceptionally old respondents had lower prevalence of overweight and obesity and lower blood pressure, thus exhibiting strong survivor phenomenon. At the same time, exceptionally old respondents from the CAHS had higher prevalence of confirmed hypertension in medical history and reported taking blood pressure medication more often. The results of this study suggest that the pattern of exceptional longevity in Croatia has a clear coastline-to-mainland gradient, which is likely to be associated with the different lifestyle and dietary patterns between these two populations. PMID- 19563163 TI - Subjective quality of life and cardiovascular risk factors in a Croatian adult population. AB - The objective of this study was to examine prevalence of cardiovascular (CV) risk factors and presence of cardiovascular diseases (CVD) in people with low subjective quality of life (SQoL) in comparison with group of people with normal SQoL. A total of 9070 participants, 2890 men and 6180 women completed a questionnaire. SQoL was assessed with single item question on satisfaction with life as a whole, and data on risk behaviors and diagnosed CVD were obtained through self-reports. Biomedical measures were administrated by public health nurse at participant's home. Results have shown that people with low SQoL have significantly more CV risk factors, and have higher prevalence of obesity and physical inactivity, all irrespective to age group. The most pronounced difference in prevalence of CV risks was found in an age group 35 to 64 years of age where people with low SQoL have significantly higher prevalence of all measured risk behaviors, and physical conditions. Participants with low SQoL perceive their general health poorer, and experience negative and depressive feelings more frequently than those with normal SQoL, in all three age groups (p < 0.001). Implications of the results have been discussed. Low SQoL was recognized as a psychological state that represents potential risk factor for the CV health. Prospective studies are needed to allow causal inferences to be drawn. PMID- 19563164 TI - Who empowers women towards healthier lifestyles? Example from western Croatia. AB - This article explores who among the doctors, other health care workers, family or somebody else most frequently advised women about their lifestyle changes related to cardiovascular health (including smoking, nutritional habits and physical activity). We analyzed who advised the most, in relation to the parameters important in the etiology of cardiovascular diseases: age, systolic blood pressure and body mass index (BMI). Sample was a part of comprehensive Croatian Adult Health Survey, comprised of women from Primorje-Goran, Istra and Lika-Senj Counties. Results indicated low frequency of advising on lifestyle changes in primary health care in all three counties, with most advice from general practitioners on nutritional habits. Family and other health care workers advised about smoking and nutrition and had strong influence in the youngest age groups. The GPs failure to counsel younger population and disease-free women could be regarded as the missed opportunity for avoidance of preventable risk factors that are associated with cardiovascular diseases. Other subjects in the health care process, as well as the family and media could fill the gaps between the patients and health care system messages. In order to create and develop such heterogeneous network approaches to training various programs and activities have to take into account all specific gender and regional characteristics. PMID- 19563165 TI - Socioeconomic inequalities estimation from public health surveys--data considerations, strengths and limitations. AB - The estimation of socioeconomic inequalities has lately become one of the major topics in public health research, as the numerous studies are showing strong association of inequalities with health status and outcomes. The aim of this study was to provide a general overview of the individual data available for the socioeconomic inequalities estimation from a public health survey and to compare different proxies used for socioeconomic inequalities estimation. The data from the Croatian Adult Health Survey were used, which is a large representative cross sectional study of the general Croatian population that was performed in 2003. Four variables were compared--education and occupation class, and objective and subjective socioeconomic estimation. Strong regional differences were recorded within Croatia, generally showing better socioeconomic indices in the capital, city of Zagreb. Although all of these variables were significantly associated to one another, breakdown into the gender stratified education-by-occupation groups indicated that income was unevenly distributed in these groups and that it had non-linear association with education, especially in the white-collared occupations. Although socioeconomic inequalities do show striking association with many health related indices and outcomes and should therefore be included in all types of research on human subjects, due attention is needed in research planning, data encoding and entry, as well as interpreting the results based on this data. PMID- 19563166 TI - [Participation of NO-synthase system in the stress-mediated reactions of the brain]. AB - Neurosis-like status developing as a result of the exposure of animals to chronic stress, which is associated with a transitory cerebral hypoxia, could cause significant structural and functional alterations in many brain structures. Realization of humoral stress effects on the brain is mediated by both extra- and intracelullar signal molecules, among which nitric oxide (NO) is considered to be one of the most potent ones. Expression of neuronal constitutive (nNOS) and inducible (iNOS) isoforms of NO-synthase was studied by immunohistochemistry in the neurons of albino rat brain after exposure of animals to chronic stress resulting in the development of neurosis-like status. Chronic stress was shown to result in the increased expression of both nNOS and iNOS in many brain areas with the predominance in neocortex and hippocampus. The administration of nonspecific inhibitor of NOS, Nomega-nitro-1-arginine methyl ester hydrochloride (L-NAME) (10 mg/kg) resulted in the aggravated depression of the animals, associated with a decrease of locomotor and exploring activities that were evaluated using the traditional tests. The application of NOS activity inhibitor caused an insignificant rise only in iNOS expression. Thus the results obtained suggest that NO is involved in the realization of stress effects with the development of a neurosis-like status. PMID- 19563167 TI - [Localization of CART-positive neurons in the amygdala and dependence of their immunoreactivity on the concentration of sex steroids]. AB - During the study of all nuclear and paleocortical structures of amygdala, the CART-peptide (cocaine-amphetamine-regulated transcript) expressing neurons were for the first time demonstrated in this region, and their immunoreactivity was shown to be influenced by sex steroids. Immunocytochemical reaction was performed on frontal slices of adult rat brain (7 female rats in estrus stage, 7 female rats in metestrus stage and 7 male rats). The portion of immunopositive neurons in animals in estrus and metestrus was counted in relation to their numbers in adjoining slices (stained using Nissl's method). The relative number of immunoreactive neurons in dorsomedial, posterior cortical nuclei and latero capsular subnucleus of the central nucleus was found to be significantly greater in estrus than in metestrus. The data obtained show that olfactory and integrative centers of amygdala may be involved in the pathogenesis of drug addiction and indicate the possibility of development of new effective methods of gene therapy with the application of an intranasal route of drug delivery. PMID- 19563168 TI - [NADPH-diaphorase activity in the motor neurons of different spinal cord segments of albino rat under normal conditions and after deafferentation]. AB - Age changes of NADPH-diaphorase activity were studied histochemically in the ventral horn motor neurons at different segmental levels of the spinal cord of rats aged 3-90 days both under normal conditions and in the model of deafferentation (by intraperitoneal capsaicin injection). Wave-like age changes of motor neuron enzyme activity were detected at the level of T(II), L(IV) and S(II) spinal segments with its increase by day 60 followed by a significant decrease to day 90. Age dynamics of NADPH-diaphorase activity development in the spinal cord motor neurons of intact rats characterizes the constructive processes in neurons, while the changes found after the deafferentation are indicative of the motor neuron damage and are manifested by an abrupt increase of the enzyme activity at the age of 90 days. PMID- 19563169 TI - [Syncytial cytoplasmic anastomoses between the neurites of caudal mesenteric ganglion cells in adult cats]. AB - Since the majority of the published data on syncytial cytoplasmic anastomoses relate to the autonomic nervous system in the early postnatal period of development, when many nerve fibers are still poorly covered by glia or have no glial sheaths at all, it was suggested that such anastomoses were not present in adults due to the significant development of glia separating individual neurites from each other. To check this assumption, we have performed an electron microscopic study of the adult cat dorsal caudal mesenteric ganglion. The cell neurites were found to be frequently covered by glial sheaths. However, almost in every sample, the syncytial pores were detected between the contacting neurites lacking glial covering layers. Sometimes serial syncytially connected neurites were seen. Axo-dendritic synapses with presynaptic perforations outside the synaptic specializations were described in the autonomic nervous system for the first time. These observations therefore provide evidence of syncytial cytoplasm connections in normal adult animals, however this does not reject the neuronal doctrine. PMID- 19563170 TI - [Nitroxidergic nerve fibers of intracerabral blood vessels]. AB - Methods of light and electron microscopic histochemistry were applied to study the structure and distribution of NADPH-diaphorase-positive neurons and processes in the parietal area of rat cerebral cortex. It was found that the most of the neurons displayed close connections with the intracerebral vessels. In the cerebral cortex, the smallest distance between the axonal plasma membrane and smooth muscle cells of the intracerebral arteries was found to be no less than 0.3-0.5 microm. Neuronal cell bodies were located in the functionally important areas of the vessels (in the areas of lateral trunk branching and in arteriolar sources), while their processes accompanied the vessels, tightly embracing them with their branches. Quite often, the neurons, the dendrites of which make contacts with the bodies or processes of over- or underlying neurons, sent their nerve fibers to the arteries, veins and capillaries. Thus, nitroxidergic neurons or their groups may control the blood flow in the different areas of vascular bed, performing the functions of the local nerve center. PMID- 19563171 TI - [Heart innervation in rats (an immunohistochemical study)]. AB - The aims of the present study were to investigate the structure of the nervous apparatus in different heart regions using the immunohistochemical methods. The study was performed in Wistar rats (n - 5). For selective staining of synaptic structures, synaptophysin (Syn) demonstration was used, while neurofilament proteins (NF-M) were used to detect axonal neurofilaments. The innervation apparatuses in the heart were shown to be unevenly distributed. High density of Syn- and of NF-M-positive axons was found in the myocardium near the atrial sinus, vena cava superior and inferior, pulmonary veins, aorta, truncus pulmonalis, as well as in the deep layers of atrial myocardium, and in the myocardium of anterior and posterior ventricle walls. In the heart apex, within the ventricular myocardium, the density of immunopositive axons was low. Using the antibodies against Syn, only the synaptic terminals of the efferent postganglionic nerve fibers and of the parasympathetic vegetative nerve fibers were selectively demonstrated in the heart, as well as the pericellular synapses on the parasympathetic neurons of intracardiac ganglia. NF-M-immunopositive axons, unlike Syn-positive terminals, were found not only in the myocardium, but also in the epicardium and the endocardium within the same regions of the heart. The authors believe that these axons belong to afferent sensory and efferent preganglionic myelinated nerve fibres of PNS. PMID- 19563172 TI - [Myocardium development in chick embryo during restriction of external respiration]. AB - The purpose of the present study was to determine the peculiarities of myocardium development in the chick embryos after closure of the half of the eggshell surface on day 11 of incubation. By the end of experimental day 3 (day 14 of incubation), myocardium demonstrated muscle cell hyperplasia and acceleration of their cytodifferentiation. These structural remodeling together with the oxygen blood capacity augmentation and increased vascularization of the intact part of the chorioallantoic membrane, compensate for the oxygen deprivation and provide for the embryo normal growth. By experimental day 7 (day 18 of incubation) the heart structural adaptive resources seem to be exhausted--the morphological signs of cardiomyocyte dystrophy, cardiac and coronary sclerosis, and marked embryo growth retardation were detected. PMID- 19563173 TI - [Formation of plasma cell infiltrates in gingiva of patients with chronic apical periodontitis]. AB - The structure of leukocyte infiltrates in gingiva of 80 patients of various ages and gender with chronic apical periodontitis was studied using light microscopy with the application of monoclonal antibodies detecting CD38 antigen. Gingival tissues of practically of all the patients with periodontitis contained 2 types of leukocyte infiltrates: infiltrates with the low plasma cell content and high numbers of neutrophils and structures with high number of plasmocytes and low concentration of neutrophilic granulocytes. Within the gingival epithelium in patients with chronic apical periodontitis CD38 + cells were absent. The histogenesis of gingival leukocyte infiltrates in chronic apical periodontitis is discussed with the special emphasis on the role of plasma cells in the development of the pathological process. PMID- 19563174 TI - [Effect of bioactive herbal preparations on structure of the gastric mucosa during stomach ulcer developing]. AB - The present study was designed to determine experimentally the effect of a biologically active phytocomposition (BAP) on the gastric mucosa structural components during the formation and healing of ulcer induced by a single intraperitoneal injection of epinephrine. Experiments were conducted using 160 Wistar albino male rats. The fundic mucosa was studied 2, 5 and 10 days after the beginning of the experiment. The integrative assessment of a gastric mucosa was given by means of a statistical matrix method under the condition of phytocorrection. BAP was shown to change the rates of cell renewal that resulted in the restoration of the dimensions of mucosal compartments. Administration of BAP decreased the degree of mucosal damage and increased the initial size of mucosal compartments by 14-15%, as compared to those at similar time intervals in animals receiving no BAP. The number of neck mucous cells in the fundic glands was stably increased. Administration of BAP provided high parameters of the general normalization index for an organ. These changes collectively favored faster restoration of the gastric mucosa integrity after an ulcer. BAP was shown to have a protective and stimulating effect on the organ structures. PMID- 19563175 TI - [Green fluorescent protein absorption and accumulation in the renal proximal tubule cells after its increased entry into blood]. AB - The uptake of green fluorescent protein (GFP) by the proximal renal tubules was studied in the anaesthetized rats using laser confocal microscopy after GFP intravenous injection or administration into the small intestine lumen. The specific green fluorescence revealed in the proximal tubule cells after intravenous injection correlated with the logarithm of GFP dose injected intravenously (r = 0.96, p < 0.05). GFP fluorescence after its intravenous injection was higher than that one after GFP infusion into the small intestine (p < 0.05). Following the increase of injected GFP dose, the epitheliocyte cytoplasm, in addition to diffuse fluorescence, demonstrated large intensely fluorescent vesicles, that was confirmed by a graphical analysis. The reported changes in the intensity and pattern of specific fluorescence indicate the enhancement of GFP absorption by the cells of proximal tubules and GFP accumulation in the intracellular compartments during its increased entry into circulation. PMID- 19563176 TI - [Comparative morphofunctional characteristics of arterial blood vessels of the different kidney segments in humans]. AB - The arterial blood vessels of 26 kidneys of people of both sexes at the first period of mature age were studied. Using Doppler renal investigation of superior segmental and anterior inferior segmental arteries in color duplex scan mode and multislice computed tomographic angiography, intravital investigation of 20 kidneys were performed. Corrosion casts of the renal vasculature of 6 kidneys, obtained at autopsy, were studied using scanning electron microscopy. It was shown that in kidneys with a single renal artery, anterior inferior and superior segments have a similar level of blood supply--when quantitative parameters of hemodynamics of these renal segments were compared, no significant differences were detected; in fact, no differences were found in the angioarchitectonics of anterior inferior and superior segments. In both segments, initial signs of age involution were found in the form of appearance of individual deformed glomeruli and some rarefaction of capillary network in the renal medulla. PMID- 19563177 TI - [Morphologic characteristic of lipid-containing and lipid-producing structures of human integument under normal conditions and in death caused by freezing]. AB - This investigation was aimed at the study of lipid-containing and lipid-producing structures of human integument under normal conditions and in death caused by freezing. The skin of interscapular and epigastral areas was studied in 105 cadavers of individuals of both sexes aged 19-83 years, who died as a result of freezing. In the control group, the skin of 10 cadavers of people killed in the car accidents and those who died by the reasons, not associated with the effect of cold, was studied. Paraffin sections were stained with hematoxylin and eosin. To demonstrate lipids, unfixed cryostat sections were stained with sudan III. The following structures are producing the skin lipids: fat cells of adipose tissue, which is localized not only in subcutaneous fat tissue (hypodermis), but forms the pads, that surround the vascular networks, secretory portions of the gland, hair follicles; keratinocytes, producing lipids as a result of terminal differentiation; a sebaceous glands containing cells (sebocytes) which transform into the sebum as a result of holocrine secretion. The lipids produced by this structures are localized either in situ or are transported to the epidermal surface. Lipid-containing structures of the skin are stratum corneum and a superficial lipid film, which is formed from the secretion of the sebaceous glands--sebum. In people who died as a result of general cooling, the content and distribution of lipids in the skin are significantly changed. In the adipose tissue, the adipocyte dimensions and number is decreased, cells without lipids are found. The thickness of the superficial lipid film was decreased as compared to that in the control group; in some areas it could be absent. In the orifices of hair follicles significant amounts of lipids are deposited. PMID- 19563178 TI - [Dynamic assessment of bone tissue osteocytic remodeling using a noninvasive method]. AB - Projectional bone mineral density (PBMD) in the proximal region of both femoral bones, the distal part of the forearm bones on both sides and in the lumbar vertebrae was studied with the method of dual energy roentgen absorption (DEXA) in two male volunteers at the age of 40 and 60 years on a daily basis during one month. On the basis of the results of the estimation of each area of interest (AI) M(av), oscillation range (4(sigma)) and the greatest difference were calculated. In addition to it, a statistic mathematic modeling was conducted through smoothing out the dynamic rows of the results achieved for each AI. It was found out that the average oscillation range exceeded 9% of the average index value, which is thrice as much as the method reproducibility error. This kind of investigation in the process of treatment allows monitoring its efficiency taking into consideration both mechanisms (osteoblastic-osteoclastic remodeling and osteocytic remodeling) as well as estimating the toxic effects of medical agents on bone cells. As a result of statistic mathematic modeling, cercaseptane periodicity of PBMD changes with the average period of 5.5 days was found. The authors see the cause of these short-term changes in osteocytic remodeling and suggest to use a dynamic successive PBMD measurements during 7-10 days as a way of non-invasive monitoring of osteocyte resorption-synthetic activity in AI. PMID- 19563179 TI - [Morphometric parameters of facial cranium asymmetry in adult man]. AB - This investigation was aimed at the evaluation of the morphometric characteristics of the contralateral sides of the facial cranium in adult men and women, the evaluation of asymmetry manifestation degree and the detection of most frequent zones of its localization. The study was conducted on 180 crania (90- male, 90--female) of the adults of I and II mature age periods. To detect the asymmetry of the facial cranium, the original "fan" principle of morphometry was developed. This approach has shown the asymmetry in the area of localization of different "fans" ("superior", "inferior", "lateral"), i.e., in the different regions of the facial cranium. On the basis of the data obtained, it can be concluded that in order to identify the asymmetry of the facial 1 cranium and to estimate the asymmetry manifestation degree it is expedient to take into account the following parameters: the distance from a nasion point to a point on the lateral edge of apertura piriformis, from a nasion point to a frontomolare temporale point, from a zygomaxillare point to a lateral point on naso-frontal suture, from a zygomaxillare point to a dacryon point, from a subspinale point to a nazomaxillare point and from a subspinale point to a lateral point on naso frontal suture. As in the series of the male crania asymmetry of the facial cranium was significantly more expressed than in the series of female ones, it is assumed to be associated with the different tone of mimic muscles. PMID- 19563180 TI - [Selective death of the striatum neurons in rats after the transient occlusion of the middle cerebral artery]. AB - The aim of the present investigation was to determine the neuronal population of rat telencephalon that was most sensitive to ischemic damage under the experimental condition of transient focal cerebral ischemia. It was found that 30 minute ischemia of the left hemisphere resulted in a significant decrease of neuronal population in the dorsomedial part of caudatoputamen. Local neural cell death was accompanied by a moderate activation of the astrocytes. PMID- 19563181 TI - [Trabeculae and intertrabecular spaces of the heart interventricular septum: anatomical structure and development]. AB - The review presents traditional and new concepts on the development of the trabeculae carneae and intertrabecular spaces (ITS) in the heart ventricles. Myocardial trabeculation preceeds the ventricular septation and at the same time underlies the formation of the interventricular septum. Trabeculae carneae seem to generate the contractile force of the embryonic myocardium. The differences between right and left ventricular trabecular patterns are conditioned by the changes of intracardiac blood flow during the prenatal period. Anatomical characteristics of the right inlet and outlet relief are reviewed. Special emphasis is given to the correlations of the terms "septomarginal trabeculation" and "moderator band". It is noticed that ITS anatomy is still insufficiently studied. The necessity to develop the anatomical criteria of normal intracardiac structures is accentuated. PMID- 19563182 TI - [First Russian anatomical dictionary (to the 225 anniversary of publication)]. PMID- 19563183 TI - [Wilhelm His Junior--discoverer of the atrioventricular bundle in the heart]. PMID- 19563184 TI - [Development of Histology in the Medical Faculty of Imperial Moscow University- Moscow Medical Academy named after I.M. Sechenov (to the 250 anniversary of the Medical Faculty Foundation)]. PMID- 19563185 TI - [The contribution of the histologists of the Military Medical Academy to the progress of tissue studies (to the 140 anniversary of the Department of Histology and Embryology of the Military Medical Academy)]. PMID- 19563186 TI - [Effects of acupuncture preventive treatment on the quality of life in patients of no-aura migraine]. AB - OBJECTIVE: To assess the therapeutic effects on acupuncture preventive treatment of no-aura migraine and its influence on the QOL (quality of life) of the patients. METHODS: Randomized controlled, double-blind and double-dummy research methods were adopted, 60 cases were randomly divided into an observation group and a control group, 30 cases in each group. The observation group was treated with acupuncture combined with oral administration of Flunarizine Hydrochloride vacuity capsules, and Baihui (GV 20), Shenting (GV 24) and Benshen (GB 13) were selected as main points. The control group was treated with oral administration of Flunarizine Hydrochloride capsules combined with acupuncture at placebo points, thrice each week, for 4 weeks. The SF-36 QOL Scale and effective rate were used for assessment of therapeutic effects before treatment, after treatment and 3 months later. RESULTS: There were significant differences in each dimension scores of SF-36 at 3 time points between the two groups (all P < 0.05). The dimension of the physiological function in the observation group was superior to that of the control group after treatment (P < 0.05), and there was no significant difference in other 7 dimensions between the two groups (all P > 0.05). After treatment and 3 months later, the effective rates were 68.0%, 68.0% in the observation group and 24.0%, 32.0% in the control group, respectively, with significant differences between the two groups (all P < 0.05). CONCLUSION: Acupuncture preventive treatment can effectively improve the life quality of the patients with migraine and reduce the migraine attack. There is no significant difference in improving the physical and psychological health of the migraine patients between acupuncture and Flunarizine Hydrochloride, and acupuncture is more effective in reducing the migraine attack days. PMID- 19563187 TI - [Clinical observation on transcutaneous electrical acupoint stimulation for treatment of functional dyspepsia]. AB - OBJECTIVE: To compare the therapeutic effects of transcutaneous electrical point stimulation and medication on functional dyspepsia. METHODS: Two hundred and fifty cases were randomly divided into an observation group and a control group, 125 cases in each group. The observation group was treated with transcutaneous electrical acupoint stimulation and oral administration of placebo, transcutaneous electrical stimulated at Zusanli (ST 36), Liangmen (ST 21), Taichong (LR 3) etc. The control group was treated with oral administration of Mosapride citrate dispersible tablets, Domperidone and Omeprazole and the placebo treatment of transcutaneous electrical acupoint stimulation, the stimulated position was 3-4 cm to the selected points of the observation group, the amount of the stimulation did not reach the treatment amount. The symptom score, the plasma motilin (MTL) concentration and the somatostatin (SS) concentration were observed before and after treatment of 3 courses. RESULTS: All symptom scores after treatment were lower than that before treatment in the two groups, the scores of the upper abdominal pain, acid regurgitation, belching and abdominal distention in the observation group were significantly lower than that in the control group (P < 0.05, P < 0.01); the treatments of both two groups can increase the plasma MTL concentration and decrease the SS concentration (both P < 0.001), and the plasma MTL concentration in the observation group was significantly higher than that in the control group (P < 0.01), and the SS concentration was significantly lower than that in the control group (P < 0.05). CONCLUSION: Transcutaneous electrical point stimulation can more reduce the symptoms of upper abdominal pain, acid regurgitation, belching and abdominal distention, etc. in the functional dyspepsia patients than medication treatment, and can increase the concentration of the plasma MTL and decrease the SS concentration, thus to improve the gastrointestinal motility. PMID- 19563188 TI - [Effect of electroacupuncture combined with auricular point tapping and pressing on serum insulin and testosterone in the patients of obese women with polycystic ovary syndrome]. AB - OBJECTIVE: To assess the therapeutic effect and mechanism of electroacupuncture combined with auricular point tapping and pressing on the obese women with polycystic ovary syndrome. METHODS: Thirty-nine cases of obese women with polycystic ovary syndrome were treated with electroacupuncture combined with auricular point tapping and pressing, body points as Tianshu (ST 25), Fenglong (ST 40), Guanyuan (CV 4) and Siman (KI 14) etc. were selected, and ear points as Kou (mouth), Wei (stomach) and Pi (spleen) etc. were selected. After 3 courses, the therapeutic effect, the body mass index (BMI), the waist circumference (WC) and the changes of the serum insulin (Ins) and testosterone (T) were compared before and after treatment. RESULTS: Of the 39 cases, 10 cases were cured, 25 cases were effective, 4 cases were ineffective, with a total effective rate of 89.7%; there were significant differences in BMI, WC, Ins and T of the patients compared with that before treatment (all P < 0.01). CONCLUSION: Electroacupuncture combined with auricular point tapping and pressing has a good clinical effect on obese women with polycystic ovary syndrome, the treatment mechanism may realized by regulating the serum insulin and the testosterone of the patients. PMID- 19563189 TI - [Clinical observation on combination of source point and Back-Shu Point for treatment of perimenopausal syndrome]. AB - OBJECTIVE: To observe the clinical therapeutic effect of combination of Source Point and Back-Shu Point on perimenopausal syndrome, and provide a new thought of therapy for perimenopausal syndrome. METHODS: Eighty cases of perimenopausal syndrome were randomly divided into a combination of Source Point and Back-Shu Point group and a routine acupuncture group, 40 cases in each group. The combination of Source Point and Back-Shu Point group was treated with acupuncture at Taixi (KI 3), Taichong (LR 3), Taibai (SP 3), Sanyinjiao (SP 6), Shenshu (BL 23), Ganshu (BL 18), Pishu (BL 20), Guanyuan (CV 4); and the routine acupuncture group was treated with acupuncture at Baihui (GV 20), Guanyuan (CV 4), Shenshu (BL 23), Taixi (KI 3), San-yinjiao (SP 6). The Kupperman score and the changes of serum estradiol (E2), follicle stimulating hormone (FSH) and luteinizing hormone (LH) were observed before and after treatment in the two groups. RESULTS: The total effective rate of 92.5% in the combination of Source Point and Back-Shu Point group was superior to 80.0% in the routine acupuncture group, with a significant difference (P < 0. 05); the Kupperman score of 24. 85 +/- 8.35 in the combination of Source Point and Back-Shu Point group was significantly decreased as compared with 35.38 +/- 9.83 in the routine acupuncture group (P < 0.05) after treatment; the contents of E2, FSH and LH in the combination of Source Point and Back-Shu Point group were significantly improved after treatment, and with a significant difference compared to the routine acupuncture group (P < 0.01, P < 0.05). CONCLUSION: Combination of Source Point and Back-Shu Point has a benign regulation function for E2, FSH and LH, and can significantly improve the Kupperman score, is superior to routine acupuncture for perimenopausal syndrome. PMID- 19563190 TI - [Observation on therapeutic effect of fire-needle therapy on lumbar intervertebral disc herniation]. AB - OBJECTIVE: To observe the therapeutic effect of fire-needle therapy on lumbar intervertebral disc herniation. METHODS: Sixty cases were randomly divided into a fire-needle group and an electroacupuncture group, 30 cases in each group. The fire-needle group was treated with fire-needle therapy and routine acupuncture, i.e., the fire-needle was given to the points on the lesion segments, and Jiaji (EX-B 2) and Ashi points above and bellow the segments, and routine acupuncture was given at Shenshu (BL 23), Zhibian (BL 54), Huantiao (GB 30), etc.; the electroacupuncture group was treated with electroacupuncture at the points as those in the fire-needle group. The scores of McGill Pain Rating Scale, Visual Analogous Scale (VAS) and present pain intensity (PPI) were compared before and after treatment for 2 weeks in the two groups. RESULTS: After treatment, all of the scores of McGill Pain Rating Scale, VAS and PPI were significantly decreased (all P < 0.01); the scores of emotion in the McGill Pain Rating Scale, VAS and PPI in the fire-needle group were significantly decreased as compared with those of the electroacupuncture group (P < 0.05, P < 0.01). CONCLUSION: Both fire needle therapy and electroacupuncture can effectively relieve pain in the patients with lumbar intervertebral disc herniation, but the former is better than the latter. PMID- 19563191 TI - [Clinical observation on therapeutic effect of different acupuncture therapies on acute peripheral facial paralysis]. AB - OBJECTIVE: To explore the better therapeutic method for acute peripheral facial paralysis. METHODS: One hundred and ninety-seven cases of acute peripheral facial paralysis were randomly divided into an observation group (n=125) and a control group (n=72). The observation group was treated by injection at acupoint combined with warming needle therapy at Yifeng (TE 17), Wangu (GB 12), Xiaguan (ST 7), Qianzheng (EX), and Taiyang (EX-HN 5). The control group was treated with electroacupuncture at Yifeng (TE 17), Yangbai (GB 14), Xiaguan (ST 7), etc. Their therapeutic effects were compared between the two groups. RESULTS: After three therapeutic courses, the cured rate and the total effective rate in the observation group were 76.0%, 100.0%, and 48.6%, 86.1% in the control group, respectively, with significant defferences between the two groups (all P < 0.01). CONCLUSION: Injection at acupoint combined with warming needle therapy could shorten the therapeutic cycle, improve the effectiveness, and decrease sequela in acute peripheral facial paralysis. PMID- 19563192 TI - [Clinical observation on effects of routine acupuncture combined with Feiteng Bafa for chloasma]. AB - OBJECTIVE: To explore the relatively efficacious acupuncture treatment for chloasma, to promote its clinical application in medical settings. METHODS: Forty cases were randomly divided into a Feiteng Bafa group (FB) and a selecting points based on differentiation of syndromes group (DS), 20 cases in each group and acupoints. Hegu (LI 4), acupoints in close proximity to chloasma and according to differentiation of syndrome were selected in DS group. The acupoints selected according to Feiteng Bafa principles were adhered in FB group. After two treatment courses, the effectiveness rate, the pro-and post-size and pro-and post color of the chloasma area were compared between the two groups. RESULTS: The size of the chloasma area decreased, and the color of the chloasma area became lighter in both groups (P < 0.01, P < 0.05), and the FB group was better than the DS group (both P < 0.05). The effectiveness rate in the FB group was 95.0%, better than that of 70.0% in the DS group. CONCLUSION: The acupuncture treatment adhered to Feiteng Bafa is better than the simple routine acupuncture for chloasma. PMID- 19563193 TI - [Clinical study on acupuncture combined with medication in restoration of gastrointestinal functions for postoperative patients with gastric cancer]. AB - OBJECTIVE: To observe clinical therapeutic effects of acupuncture combined with medication in restoration of gastrointestinal functions for postoperative patients with gastric cancer. METHODS: Ninety patients undergoing radical surgeries for gastric cancer were randomly, according to the sequence of their operations, divided into three groups: a control group treated conventionally after their surgeries (group CONT, 30 cases), a Chinese medicine group treated by Simo Decoction administered by way of a nutrient canal in addition to the conventional treatment (group CM, 30 cases), and an acupuncture plus Chinese medicine group treated by warming needling in addition to those given in the Chinese medicine group (group ACUP+CM, 30 cases). Therapeutic effects were estimated 10 days after their operations. RESULTS: The time for restoration of gastrointestinal functions was obviously shortened, and the problems of poor appetite and difficulty in defecation were more markedly improved in group ACUP+CM than those in both group CONT and group CM (P < 0.01, P < 0.05). Ten days after operations, the number of patients with normal lymphocytes and normal percentage rate of lymphocytes to neutrophile granulocytes was obviously more in group ACUP+CM than those in both group CONT and group CM (P < 0.01, P < 0.05). CONCLUSION: Acupuncture combined with Chinese medicine is favorable in accelerating early air exhaustion and defecation, improving clinical symptoms, as well as in bi-directional regulating peripheral white blood cells. PMID- 19563194 TI - [Analysis of characteristics of meridians and acupoints selected for treating migraine in past dynasties based on data excavation]. AB - OBJECTIVE: To analyze the regularity of selection of meridians and acupoints for treating migraine in past dynasties. METHODS: The data were collected from 178 kinds of literature concerning acupuncture and moxibustion from the Pre-Qin Dynasty to the end of the Qing Dynasty, based on which bibliographic database in ancient acupuncture and moxibustion for treating migraine was established. By way of data excavation and analysis, meridians and acupoints and their combinations frequently used for the treatment in successive dynasties were screened, and the potential regularity was searched. RESULTS: In the treatment of migraine, the Gall Bladder Meridian was the meridian used most frequently, and both local and remote points in the four extremities were used. Acupoints in yang meridians, especially Fengchi (GB 20), Sizhukong (TE 23) and Shuaigu (GB 8) in Shaoyang meridians were used most frequently. In terms of classification of the special points selected, besides the Crossing Points with meridian qi passing through and crossing in the head, points below the elbow and knee joints, such as Source points, Luo Points, Eight Confluence Points, as well as Five Shu Points were also much frequently used. CONCLUSION: According to data excavation and analysis of references recorded on the treatment of migraine by acupuncture and moxibustion in past dynasties, it is concluded that meridian theories are the foundation and prerequisite for guiding clinical treatment and point selection, in which special acupoints are the main components and the core in point prescription. PMID- 19563195 TI - [On the characteristics of acupoints composed by trinity]. AB - In the present paper, the characteristics of acupoints in composition and their functional activities are discussed based on related domestic and abroad investigated results achieved in this area. Niche microenvironment of adult stem cells and correlative special differentiated cells are assembled, like gathering in caves, under the complex ation of network junctions of biological stress. A hypothesis of trinity is put forward to explain the construction and functional activities of acupoints. PMID- 19563196 TI - [Relationship between laser acupuncture analgesia and the function of mast cells]. AB - OBJECTIVE: To observe the analgesic effects of single-and combined-laser irradiation with low-intensity applied at "Zusanli" (ST 36) in rats, and their relation to degranulation of mast cells. METHODS: Sixty-six SD rats were randomly divided into 6 groups: normal control group (Group NC), model control group (Group MC), sham irradiation group (Group SI), 10.6 microm laser irradiation group (Group 10.6 microm LI), 650 nm laser irradiation group (Group 650 nm LI) and combined (10.6 microm + 650 nm) laser irradiation group (Group CLI). Complete Freund's Adjuvant (0.05 mL) was injected into the left ankle joints of all the rats except those in Group NC to cause acute adjuvant-induced arthritis. In treatment, laser irradiation was applied at "Zusanli" (ST 36) for 30 minutes in all the rats except those in Group NC and Group MC. The paw withdrawal latency (PWL) to radian heat was used to compare analgesic effects among the groups. By means of toluidine blue, dyed slices of local tissues of "Zusanli" (ST 36) were used to observe changes of mast cell degranulation before and after laser irradiation. RESULTS: The pain thresholds to irradiation of the rats in Group 650 nm LI and Group CLI were significantly higher than those in Group MC and Group SI (P < 0.01), and the mast cell degranulation rate in Group 650 nm LI and Group CLI were also significantly higher than that in Group MC and Group SI (P < 0.001). The pain threshold and mast cell degranulation rate in Group 10. 6 microm LI were not significantly different from those in Group MC and Group SI. There was a linear correlation between mast cell degranulation rate and PWL with 0. 737 in coefficient (P < 0.001). CONCLUSION: Single 650 nm laser and combined 650 nm + 10.6 microm laser with low intensity irradiated at "Zusanli" (ST 36) in acute adjuvant rats can provide remarkable analgesic effects, and there was a positive correlation between mast cell degranulation rate and analgesic effects, which plays an important part in laser irradiation-induced analgesia. PMID- 19563197 TI - [Experience of professor Sun Liuhe in treating facial peripheral paralysis]. AB - Professor Sun Liuhe is engaged in medical service for over 40 years. He is deeply involved in research on intractable and complicated diseases. Especially, in treating facial paralysis, he makes diagnosis and treatment based on overall analysis of symptoms and causes, differentiation of syndrome for etiology. Without confining himself to ancient treatment methods, by applying both acupuncture and Chinese medicine, selecting auxiliary acupoints according to differentiation of meridians related to illness, as well as considering anatomy. Professor Sun holds ancient and modern therapeutic methods, brings forth ideas of using new acupoints to prevent perversion based on pulse tracings, and uses cutting therapy for the cases suffering from facial paralysis for a long time. This method can shorten treatment courses, to a great extent, and achieve good therapeutic effects on intractable facial paralysis, which has revealed distinctness of acupuncture therapy. PMID- 19563198 TI - [Retrospective analysis of Mr. Xie Xiliang's medical records accumulated in 30 years on direct moxibustion for treating hepatitis B]. AB - To analyze retrospectively Mr. Xie Xiliang's medical records accumulated in 30 years on direct moxibustion for treating hepatitis B, in which 86 cases were included. Before accepting direct moxibustion, 80.23% of the cases had been treated by conventional Chinese and western medicine, but no satisfactory therapeutic effects were obtained. Direct moxibustion was applied for 7-9 cones at each point selected. Point selection was related to ages. Usually, only a few points were used, and Ganshu (BL 18) and Pishu (BL 20) were used as main points. Shenzhu (GV 12) was added for children, and Zusanli (ST 36) added for adults. The improvement rate in clinical symptoms and physical signs were 100%. In the cases with hepatauxe, splenauxe, liver cirrhosis, and abdominal dropsy diagnozed by the B-ultrasonic examination, the improvement was obvious. Of the total, the rates turning to be negative in HBsAg, HBeAg and HBcAb were 28.85%, 38.46%, and 36.54%, respectively. It took some 10 days to 1 month to begin to show improvement in physical signs and hepetic functions, and about 3-6 months for their restoration. For the five items tested in type B hepatitis, it took 5 months to 1 year, even several years, to show their improvement. It is indicated that direct moxibustion is definitely effective in treating type B hepatitis. It is easy to handle and worth popularizing. PMID- 19563199 TI - [The fundamental characteristics and application of psychological intervention on acupuncture therapy]. AB - The process of acupuncture therapy is a complete combination of linguistic suggestion, cognitive behavioral therapy and body treatment systems. Differentiation of syndrome and diagnosis play the role of linguistic suggestion, while the magnified phenomenon of bio-information and possible manipulation on the arrival of qi play the role of cognitive behavioral therapy. The objective effectiveness of acupuncture not only includes clinical treatment, but also contains reducing or preventing foreign malignant psychological stimulation, regulating the concentration of 5-hydroxytryptamine and dopamine, and keeping the inter environment stable etc. According to the process of treating patient as followed with "telling his sickness, establishing his confidence, inducing his feeling and relieving his suffering", treatment is carried out with taking the arrival of qi as the key point, combining the steps of characteristics of psychological treatment effectively, and cooperating with psychological and body treatments to obtain effectiveness. It accords with Chinese medical theories of simultaneous treatment of the branch and root as well as effectiveness following arrival of qi. PMID- 19563200 TI - [Strengthening innovation in clinical research methodology of acupuncture and moxibustion to promote internationalization process of acupuncture-moxibustion]. AB - In order to explore the problems and countermeasure in the methodology of acupuncture and moxibustion clinical researches at present, clinical research literatures about acupuncture and moxibustion (Acup-Mox) published in recent years in our country were reviewed. For the urgent need of the current internationalization of Acup-Mox, the authors proposed the model of clinical research on Acup-Mox, which should strictly stick to the international standard and fully embody traditional Chinese medicine characteristics in the intervention measures of acupuncture. It is indicated that innovation of the methodology about clinical researches of Acup-Mox has great significance in improving the quality of clinical research on Acup-Mox in our country and promoting internationalization of Acup-Mox. PMID- 19563201 TI - [Discussion about the divided methods and unification on the location of scalp acupuncture]. AB - Two kinds of divided methods currently used in clinic and their unified new method on the scalp acupuncture were discussed. Through comparing with the nomination and selecting methods on the stimulating location, the characteristics of the two kinds of divided methods on the location of scalp acupuncture were pointed out respectively. The advantage of the method with scalp projection areas is that the nomination is easy to associate and remember, however its disadvantage is that the measurement on point-selecting is less reasonable. For the method with standard treatment lines, its measurement is more reasonable, but its nomination on the location is sophisticated and difficult to be associated and remembered, especially having more repeated lines. The new divided method of scalp acupuncture combines the advantages of the two methods in order to easily remember and freely apply through their unification, which is worth being popularized. PMID- 19563202 TI - [Research on Tuina treatment for postpartum hypolactation]. AB - In this paper, the TCM typing of postpartum hypolactation is introduced and the manipulation and duration of Tuina treatment for postpartum hypolactation are summarized, and the effectiveness index used in Tuina treatment is proposed. It is held that the Tuina therapy will be widely used for treatment of the postpartum hypolactation, and the convenient and effective Tuina manipulation will promote the development of postpartum nursing and will be used in gynecology department and enlarge the therapeutic range of the Tuina therapy. PMID- 19563203 TI - [Analysis on acupuncture literature in Science Citation Index (SCI) periodicals in 2007]. AB - To grasp the international developing tendency of acupuncture research and provide some references for promoting acupuncture and moxibustion internationalization process, the articles about acupuncture in Science Citation Index (SCI) periodicals in 2007 were retrieved by adopting the retrieval tactics on line in combination with database searching. Results indicate that 257 articles about acupuncture had been retrived from the SCI Web databases. These articles were published in 125 journals respectively, most of which were Euramerican journals. Among these journals, the impact factor of the Journal of the American Medical Association (JAMA), 25. 547, is the highest one. It is shown that the impact factors of the SCI periodicals, in which acupuncture articles embodied are increased, the quality of these articles are improved obviously and the types of the articles are various in 2007, but there is obvious difference in the results of these studies due to the difference of experimental methods, the subjects of these experiments and acupuncture manipulations. Therefore, standardization of many problems arising from the researches on acupuncture is extremely imminent. PMID- 19563204 TI - [Standardized manipulations of sharp hooked needling for treating migraine and investigation on its mechanisms]. AB - A comparison of recognition to migraine and treatment methods for it was made between traditional Chinese medicine and western medicine in this paper. Emphasis was made on standardized manipulation processes and techniques of sharp hooked needling for treating the disease, based on both the organic conception of the human body, the theory held by TCM, and on the principle of treatment according to pathogenesis obtained through differentiation of symptoms and signs. The necessary preparation before operations, methods for its application and announcements in the standardized manipulations of sharp hooked needling were all explained in detail. Based on its needling, pricking, dissecting and relaxing effects, the mechanisms of sharp hooked needling in treating the disease were investigated. It is concluded that sharp hooked needling is very effective in treating migraine. PMID- 19563205 TI - [Systematic evaluation of the randomized controlled trials about acupuncture and moxibustion treatment of allergic rhinitis]. AB - OBJECTIVE: To evaluate the clinical effect and safety of acupuncture and moxibustion treatment for allergic rhinitis and to analyze the present situation of clinical researches. METHODS: A search in PubMed, Cochrane Library, Chinese Biology Medicine (CBM) disk, and China National Knowledge Infrastructure (CNKI) databases was performed to gather the randomized controlled trials about acupuncture and moxibustion treatment for allergic rhinitis, identify additional clinical trials met the inclusion criteria and measure their qualities by using Cochrane Reviewers' Handbook 5.0. Statistical analysis was carried out by RevMan 4.2.8. RESULTS: A meta-analysis was performed on a total of 1076 patients involved in 12 papers which met the inclusion criteria. There were significant differences in both cure rate (Incorporate RR = 1.86, 95% CI 1.51, 2.29, Z = 5.82, P < 0.00001) and marked improvement rate (Incorporate RR = 1.58, 95% CI 1.32, 1.89, Z = 4.94, P < 0.00001) between acupuncture and moxibustion treatment and the routine medicine treatment for allergic rhinitis. CONCLUSION: Acupuncture and moxibustion to treat allergic rhinitis is effective and safe and may have certain advantage over the routine medicine treatment. However, as for the low quality of partial inclusion literatures, no definite conclusion can be obtained as yet and it still waits for higher quality researches to further prove the dominance of acupuncture and moxibustion treatment for allergic rhinitis. PMID- 19563206 TI - Interpretation and considerations on the safety evaluation of human drug metabolites. AB - The final Food and Drug Administration guidance on the safety testing of drug metabolites was published in February 2008. Discussions of the role and applications of this guidance were addressed at several public scientific meetings over the past year. One of the main differences between the draft and the finalized guidance is that in the latter, the human metabolite level was correlated to the parent drug level in plasma, whereas this parameter was considered in relationship to administered dose or total exposure in the draft guidance. The parent drug concentration in plasma has traditionally been the parameter commonly measured in animals during drug development and the one used to estimate drug clinical levels and to assess human risk. Moreover, circulating parent drug in general is the molecule with the intended therapeutic and pharmacologic effect. Therefore, it is appropriate to compare metabolite concentration to that of the parent drug. This report elaborates on this issue and supports other alternative rational and scientific approaches on the design of nonclinical studies that may be needed to test a human drug metabolite. PMID- 19563207 TI - Reverse type I binding spectra of human cytochrome P450 1B1 induced by flavonoid, stilbene, pyrene, naphthalene, phenanthrene, and biphenyl derivatives that inhibit catalytic activity: a structure-function relationship study. AB - Fifty-one chemicals including derivatives of 16 flavonoids, three stilbenes, six pyrenes, seven naphthalenes, seven phenanthrenes, 10 biphenyls, 17beta-estradiol, and estrone were examined for their abilities to induce reverse type I binding spectra with human cytochrome P450 (P450) 1B1 and to inhibit 7-ethoxyresorufin O deethylation (EROD) activities catalyzed by P450 1B1. Forty-nine chemicals showed reverse type I spectra with P450 1B1, and we found that 3,5,7-trihydroxyflavone, 3',4'-dimethoxy-5,7-dihydroxyflavone, 4'-methoxy-5,7-dihydroxyflavone, alpha- and beta-naphthoflavones, 2,4,3',5'-tetramethoxystilbene, pyrene, and several acetylenic pyrenes and phenanthrenes were strong inducers of the spectra and also potent inhibitors of EROD activities catalyzed by P450 1B1. The spectral dissociation constant (K(s)) and the magnitude of the binding (DeltaA(max)/K(s)) of 49 chemicals were correlated with the inhibition potencies of EROD activities by these chemicals [correlation coefficients (r) of 0.72 and 0.74, respectively]. The K(s) and DeltaA(max)/K(s) values were more correlated with IC(50) values when compared in a group of derivatives of flavonoids, stilbenes, and estrogens (r = 0.81 and 0.88, respectively) or a group of derivatives of pyrenes, naphthalenes, phenanthrenes, and biphenyls (r = 0.88 and 0.91, respectively). Among 14 flavonoids examined, 3,5,7-trihydroxyflavone and 4'-methoxy- and 3',4'-dimethoxy 5,7-dihydroxyflavone were more active than flavone in interacting with P450 1B1, but the respective 7,8-dihydroxyflavones were less active. Pyrene itself was highly active in interacting with P450 1B1, but its binding was slightly decreased when substituted with acetylenic groups. In contrast, substitution of naphthalene with methyl and ethyl propargyl ethers led to more interaction with P450 1B1 than with naphthalene itself. Similarly, substitution on phenanthrene or biphenyl with acetylenic groups and propargyl ethers increased affinities to P450 1B1. These results suggest that the reverse type I binding of chemicals to P450 1B1 may determine how they interact with and inhibit the catalytic activity of the enzyme. Substitutions on the compounds with various acetylenic groups and propargyl ethers cause an increase or decrease of their affinities to P450 1B1, depending on the parent compound used. PMID- 19563209 TI - Oxidation of 4-chloroaniline studied by on-line electrochemistry electrospray ionization mass spectrometry. AB - The oxidation of 4-chloroaniline (4-CA) has been studied by electrochemistry (EC) coupled on-line with electrospray ionization mass spectrometry (ESI-MS) using two electrochemical flow cells of different design. The experimental results, which generally verify previously suggested oxidation pathways for 4-CA, also indicate the presence of an up to now unrecognized comproportionation reaction. The oxidation of 4-CA (m/z 128.2) was found to give rise to the formation of both an oxidized dimer, 4-[(4-chlorophenyl)imino]-2,5-cyclohexadien-1-imine (m/z 217.2), and a reduced dimer, 4-amino-4'-chlorodiphenylamine (m/z 219.2), in addition to a dimer intermediate (m/z 253.2). The unexpected formation of the reduced dimer is shown to stem from a comproportionation reaction involving 4-CA and the oxidized dimer. The presence of the latter reaction was clearly seen by comparing results obtained with two thin-layer flow cells, both with conversion efficiencies of 50% under mass transport controlled conditions but of different design with respect to the influence of the counter electrode reaction on the reaction at the working electrode. The experimental results demonstrate that the formation of the reduced dimer is favored by a decrease in the local pH in the flow cell, and the influence of the pH on the oxidation of 4-CA was also investigated in the pH range between 2.0 and 6.0 using off-line voltammetry. It is concluded that EC/ESI MS is a powerful tool for the study of the present type of reactions and that studies of the reaction pathways of these systems are best carried out under noncoulometric experimental conditions as the latter facilitates the detection of reaction intermediates. Comproportionation reactions, similar to the reaction present in the 4-CA system, can also be expected to be present during the formation of conducting polymers such as polyaniline and polypyrrole. PMID- 19563208 TI - Down-regulation of 14-3-3 isoforms and annexin A5 proteins in lung adenocarcinoma induced by the tobacco-specific nitrosamine NNK in the A/J mouse revealed by proteomic analysis. AB - The tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a potent lung carcinogen in the A/J mouse model. Here we identified and validated, using two-dimensional difference gel electrophoresis (2D-DIGE) coupled with mass spectrometry and immunoblotting, proteins that are differentially expressed in the lungs of mice treated with NNK versus vehicle control treatment. We also determined whether protein levels in the lungs of NNK-treated mice could be further modulated by the chemopreventive agent 1,4 phenylenebis(methylene)selenocyanate (p-XSC). The proteins identified in this study are SEC14-like 3, dihydropyrimidinase-like 2, proteasome subunit alpha type 5, annexin A5, 14-3-3 protein isoforms (theta, epsilon, sigma, and zeta), Rho GDP dissociation inhibitor alpha, myosin light polypeptide 6, tubulin-alpha-1, vimentin, Atp5b protein, alpha-1-antitrypsin, and Clara cell 10 kDa protein (CC10). Among those proteins, we demonstrated for the first time that 14-3-3 isoforms (theta, epsilon, and sigma) and annexin A5 were significantly down regulated in mouse lung adenocarcinoma induced by NNK and were recovered by p XSC. These proteins are involved in a variety of biological functions that are critical in lung carcinogenesis. Identification of these proteins in surrogate tissue in future studies would be highly useful in early detection of lung adenocarcinoma and clinical chemoprevention trials. PMID- 19563211 TI - Design, implementation, and field testing of a portable fluorescence-based vapor sensor. AB - The design and implementation of a portable fluorescence-based vapor sensing system are described. The system incorporates previously developed microsensor array technology into a compact, low-power device capable of collecting and delivering ambient vapor samples to the array while monitoring and recording the fluorescent responses of the sensors. The sensors respond differentially when exposed to a sample vapor and, when processed using a support vector machine (SVM) pattern recognition algorithm, are shown to discriminate between three classes of petroleum distillates. The system was characterized using sample vapors prepared under several different conditions in three sensing scenarios. The first scenario demonstrates the basic operational capability of the device in the field by presenting high concentration vapors to the array. The second scenario introduces the potential for a greater degree of variability in both sample vapor concentration and composition in an effort to emulate real-world sensing conditions. The third scenario uses an on-board trained pattern recognition algorithm to identify unknown vapors as their responses are collected. The device demonstrated high classification accuracy throughout the field tests and is capable of improving its classification accuracy when challenged with samples presented under variable ambient conditions by enhancing the signal-to-noise ratio of the array response. PMID- 19563210 TI - Antigen peptide-based immunosensors for rapid detection of antibodies and antigens. AB - The homogeneous immunosensor design described here utilizes the bivalent nature of the antibody. Antigen peptide is conjugated using flexible linkers with short complementary oligonucleotides (signaling oligonucleotides), each of which containing a fluorochrome that can form a fluorescence resonance energy transfer (FRET) donor-acceptor pair. The complementary signaling oligonucleotides are short enough to prevent their annealing on their own. Binding of the peptide signaling oligonucleotide constructs to bivalent antibody results in a large increase in local concentration of signaling oligonucleotides causing their annealing and appearance of FRET signal. We used simple model system (antibiotin antibody) to obtain proof-of-principle validation of the sensor design. We then constructed two sensors based on two peptides corresponding to the antigens of two antibodies raised against human cardiac troponin I. We demonstrated that these sensors could be used for sensitive detection of the antibody and for competition-based detection of the intact troponin I. Furthermore, we showed that these sensors could be used for detection of kinase activity targeting the antigen peptide. These simple and robust immunosensors may find applications in antibody detection (for example, in diagnosis of autoimmune or infectious disease), in protein detection (especially when speed of detection is essential), and in assays for detecting enzymatic activities involved in post-translational modifications of proteins. PMID- 19563212 TI - Electrostatic sample nebulization for improved sample vaporization in the split/splitless gas chromatography inlet. AB - The split/splitless inlet system has basically the same fundamental drawbacks it had when it was introduced: poor repeatability of the injected amount of sample and discrimination of high-boiling analytes. Hot needle injection improves the repeatability of the sample transfer but suffers from in-needle discrimination. Injection with a fast autosampler, resulting in minimal heating of the needle, solves this problem but usually requires a glass wool packing in the inlet liner to assist in vaporization of the sample. As glass wool has been reported to cause degradation of labile analytes, it cannot be applied as a general remedy for improving incomplete vaporization. In this paper, a novel concept, based on electrostatic nebulization of the injected sample, is presented. The resulting fine droplets promote a more effective heat transfer and a rapid vaporization. Evaluation of the electrospray inlet in the split mode, using a straight, empty glass liner and a cold needle, showed an improvement in peak area repeatability by about 1 order of magnitude, compared with the results obtained when no electrostatic field was applied. Splitless injection of a series of hydrocarbons up to C(28) in the electrospray inlet with an empty, tapered liner, using a cold needle, showed no measurable analyte discrimination. The relative standard deviation in terms of area count for the largest hydrocarbon (C(28)) was <1.5%, compared to approximately 30% for injections where no high voltage was applied. PMID- 19563213 TI - Macromolecular scaffolds for immobilizing small molecule microarrays in label free detection of protein-ligand interactions on solid support. AB - We explored two macromolecular scaffolds, bovine serum albumin (BSA) and polyvinyl alcohol (PVA), as chemically complementary platforms for immobilizing small molecule compounds on functionalized glass slides. We conjugated biotin molecules to BSA and amine-derivatized PVA and subsequently immobilized the conjugates on epoxy-functionalized glass slides through reaction of free amine residues on BSA and PVA with surface-bound epoxy groups. We studied binding reactions of such immobilized small molecule targets with solution-phase protein probes using an oblique-incidence reflectivity difference scanning optical microscope. The results showed that both BSA and amine-derivatized PVA were effective and efficient as carriers of small molecules with NHS residues and fluoric residues and for immobilization on epoxy-coated solid surfaces. A significant fraction of the conjugated small molecules retain their innate chemical activity. PMID- 19563215 TI - Capillary electrophoresis-assisted identification of peroxyl radical generated by single-walled carbon nanotubes in a cell-free system. AB - The mechanism of how single-walled carbon nanotubes (SWCNTs) induce oxidative stress in cells has always been under debate. In the present paper, we have clearly identified the generation of peroxyl radical (ROO(*)) by the unmodified SWCNT and the poly(ethylene glycol)-functionalized SWCNT (SWCNT-PEG) in aqueous solutions using a simple analytical technique: capillary electrophoresis (CE). CE provides a sensitive detection platform for the oxidation product of the employed reactive oxygen species (ROS) indicator, 2,7-dichlorodihydrofluorescein (H(2)DCF). In addition, its high separation power eliminates the fluorescence quenching exerted by SWCNTs, which was a big problem that has been encountered in measurements with microplate readers, and facilitates the screening of a large variety of ROS scavengers, including small molecules, surfactant, and proteins, in our study. Furthermore, CE enables the simultaneous incubation of multiple ROS indicators for comparison of their oxidation efficiency, improving the detection specificity. The adsorption of oxygen on the SWCNT surface could be the source of the identified ROO(*), because surface coverage by surfactant or proteins could inhibit the oxidation and the anaerobic experiment led to data agreeing with the Langmuir isotherm. The peroxyl radical can serve as the seed for the production of more-active ROS after the SWCNT enters the cell and, therefore, SWCNTs hold potential toxicity to the biological hosts. PMID- 19563214 TI - Measurement of deuterium-labeled phylloquinone in plasma by high-performance liquid chromatography/mass spectrometry. AB - Phylloquinone (vitamin K(1)) is a lipophilic compound present in plasma at low concentrations, which presents technical challenges for determining its bioavailability or metabolic fate using stable isotopes. We developed a method to simultaneously measure unlabeled and deuterium-labeled phylloquinone concentrations in plasma specimens using high-performance liquid chromatography/mass spectrometry with atmospheric pressure chemical ionization (LC-APCI/MS). Phylloquinone was extracted from plasma using hexane, further purified by solid-phase extraction, and then quantified using high-performance liquid chromatography with an APCI/MS as a detector. Plotting the expected versus the measured amount of serial dilutions of either unlabeled or labeled phylloquinone gave correlation coefficients (R) of 0.999 for both compounds. The minimum detectable concentrations of unlabeled and labeled phylloquinone were 0.05 and 0.08 pmol/injection, respectively. Pooled plasma samples spiked with between 0.5 and 32 nmol phylloquinone/L gave average recoveries of 96.7% with 5.4% relative standard deviation (RSD) for unlabeled phylloquinone and 96.2% with 6.6% RSD for labeled phylloquinone. Plasma phylloquinone concentrations determined by LC-fluorescence and LC-APCI/MS methods from healthy subjects (n = 17) were not statistically different (P = 0.13). The LC-APCI/MS method is a sensitive technique for simultaneous determination of both unlabeled and labeled phylloquinone and can be applied to bioavailability studies. PMID- 19563216 TI - C2AB: a molecular glue for lipid vesicles with a negatively charged surface. AB - Artificial particulate systems such as lipid vesicles are found in a variety of biomedical applications such as drug delivery and targeting. More versatile layers of control would be added if liposomes could be glued together on demand while stabilized against fusion. Here, we present a two-component molecular glue composed of a protein and calcium ions, with each component specialized for fast and specific binding to negatively charged lipid membranes. Upon mixing the two components, the high affinity binding of this glue starts to tightly bridge two lipid vesicles on a subsecond scale. Furthermore, highly charged liposomes are beneficial in preventing spontaneous fusion before applying the molecular glue. PMID- 19563217 TI - Surface rheology of trisilanolisobutyl--POSS at the air/water interface. AB - Trisilanol polyhedral oligomeric silsesquioxane (POSS) derivatives have recently been reported as a new class of amphiphilic molecules that form stable Langmuir monolayers at the air/water interface and exhibit interesting phase transitions and aggregation behavior. In this letter, surface light scattering (SLS) has been used to probe the dilational viscoelastic behavior of trisilanolisobutyl-POSS in the monolayer regime of its surface pressure-area per molecule isotherm at 22.5 degrees C. Results showed that the static dilational elastic modulus is in excellent agreement with the dynamic dilational elastic modulus in the monolayer regime over the limited wavevector range of 404.3-512.6 cm(-1). The films possessed a moderate maximum dynamic dilational elastic modulus (approximately 50 mN.m(-1)) prior to film collapse. Moreover, the films exhibited small surface viscosities such that the films are almost purely elastic over the frequency range studied throughout the monolayer regime. PMID- 19563218 TI - Assembly of self-assembled monolayer-coated Al2O3 on TiO2 thin films for the fabrication of renewable superhydrophobic-superhydrophilic structures. AB - A renewable superhydrophobic-superhydrophilic pattern with a minimum dimension of 50 microm is prepared from octadecyltrimethoxysilane self-assembled monolayer covered superhydrophobic Al2O3 overlayers on a superhydrophilic TiO2 surface via self-assembly and calcination of boehmite (AlOOH.nH2O) particles. The resulting Al2O3 layer plays dual roles as a superhydrophobic layer and as a UV-blocking layer for the underlying TiO2. PMID- 19563219 TI - Self-assembly of metal oxide nanoparticles into hierarchically patterned porous architectures using ionic liquid/oil emulsions. AB - Hierarchically patterned macroporous TiO2 structures can be fabricated via the spontaneous self-assembly of TiO2 nanoparticles prepared using a mixture of 1 octadecene (ODE) and an ODE-immiscible 1-alkyl-3-methylimidazolium-based ionic liquid as the reaction medium. A study of the influence of side chain lengths of ionic liquids (n=4, 8, or 16) reveals that this parameter can be further used to fine-tune the morphologies of the products. This synthetic methodology can also be extended to the formation of patterned macroporous ZrO2 and Fe3O4 structures. Finally, the potential reasons for the formation of hierarchical structures are discussed and the implications to further research are proposed. PMID- 19563220 TI - Room temperature one-step polyol synthesis of anatase TiO2 nanotube arrays: photoelectrochemical properties. AB - We describe an optimized anodization process to fabricate anatase TiO2 nanotube arrays at room temperature using a polyol electrolyte. The critical roles of water and anodization voltage are investigated, and mechanistic actions are considered. The as-fabricated electrodes show low efficiency when used as photoanodes to photoelectrochemically split water; however, their efficiency is remarkably enhanced upon their annealing at temperatures as low as 300 degrees C. The efficiency of the annealed samples was found to increase with the anatase content in the as-anodized electrodes, suggesting that these pre-existing crystallites can act as seeding layers that enhance the nucleation and growth of further anatase crystallites. PMID- 19563221 TI - Catanionic surfactants as nanospring suspensions: a model. AB - A model for a dilute suspension of nanosprings, whose equilibrium configuration and extension are controlled by electrical double layer forces, is presented along with a model for changing packing parameter. The dependence of viscosity on surface charge is calculated. The possibility of shear-thickening is demonstrated. It is argued that mixtures of cationic and anionic surfactants which show two peaks in viscosity and a minimum along with shear-thickening are likely candidates for this model. PMID- 19563222 TI - The impact of electrolyte on the aggregation of the complexes of hyperbranched poly(ethyleneimine) and sodium dodecyl sulfate. AB - The aggregation of the negatively charged complexes of hyperbranched poly(ethylenimine) (PEI) and sodium dodecyl sulfate (SDS) has been investigated at different sodium chloride (NaCl) concentrations using coagulation kinetics, electrophoretic mobility and dynamic light scattering measurements. The observed variation of the initial rate of coagulation with NaCl concentration indicates the formation of kinetically stable colloid dispersions in the investigated composition and pH range. These dispersions are electrostatically stabilized due to the adsorption of excess dodecyl sulfate ions on the surface of the polyelectrolyte/surfactant particles. Because of the enhanced adsorption of the anionic surfactant, the kinetic stability of the PEI/SDS dispersions increases with increasing SDS concentration and decreasing pH. Finally, we rationalize the effect of salt on the phase behavior and surface properties of polyelectrolyte/surfactant mixtures in terms of the salt-induced aggregation features of polyelectrolyte/surfactant particles. PMID- 19563223 TI - Monte Carlo simulations of small vesicles under osmotic pressure. AB - A simple model for amphiphilic molecules is sufficient to predict self-assembly into spherical vesicles. The model is also useful to study the influence of osmotic pressure on the shape of vesicles, which serve as carriers in biological cells. The stability of small vesicles subjected to relatively high osmotic pressures is demonstrated. Fluctuations of small vesicles under osmotic stress are found to be in semiquantitative agreement with a macroscopic description. Small deviations between simulated results for vesicle fluctuations and macroscopic elasticity theory could result from the relative large membrane thickness in comparison to the vesicle radius. The simulations demonstrate that an osmotic pressure exerted by solute molecules outside a spherical vesicle can cause a shape transition, in agreement with results based on the elasticity theory of membranes. PMID- 19563224 TI - A geometric model for the dynamics of microchannel emulsification. AB - Microchannel emulsification is an interfacial tension driven method to produce monodisperse microdroplets, or microspheres. In this paper we introduce a model for describing the dynamics of microchannel emulsification based on simple time dependent geometric shape analysis. The model is based on mechanistic principles that simultaneously predicts both process and microchannel geometry effects. The model contains no adjustable (fit) parameters and is thus fully predictive for oil in water emulsification. The model is easy to use and does not require extensive computational time and/or memory. The model was validated by comparison with the experimental results published by Sugiura and co-workers and we found excellent agreement. It was found that the droplet size of oil in water emulsions could be fully predicted using only two dimensionless numbers, an adapted capillary number that also comprises effects of terrace width and height, and the ratio of terrace length over terrace height. Based on these findings, a dimensionless design map could be constructed for a wide range of process conditions and microchannel dimensions. PMID- 19563225 TI - Accurately tuning the charge on giant polyoxometalate type Keplerates through stoichiometric interaction with cationic surfactants. AB - We report an approach of exploring the interaction between cationic surfactants and a type of structurally well-defined, spherical "Keplerate" polyoxometalate (POM) macroanionic molecular clusters, {Mo72V30}, in aqueous solution. The effectiveness of the interaction can be determined by monitoring the size change of the "blackberry" supramolecular structures formed by the self-assembly of {Mo72V30} macroions, which is determined by the effective charge density on the macroions. Long-chain surfactants (CTAB and CTAT) can interact with {Mo72V30} macroions stoichiometrically and lower their charge density. Consequently, the blackberry size decreases continuously with increasing surfactant concentration in solution. On the other hand, for short-chain surfactants (e.g., OTAB), a larger fraction of surfactants exist as discrete chains in solution and do not strongly interact with the macroions. This approach shows that a controllable amount of suitable surfactants can accurately tune the charge on large molecular clusters. PMID- 19563226 TI - Solvent dependence of the activation energy of attachment determined by single molecule observations of surfactant adsorption. AB - Single-molecule total internal reflection fluorescence microscopy was used to obtain real-time images of fluorescently labeled hexadecanoic (palmitic) acid molecules as they adsorbed at the interface between fused silica and three different solvents: hexadecane (HD), tetrahydrofuran (THF), and water. These solvents were chosen to explore the effect of solvent polarity on the activation energy associated with the attachment rate, i.e., the rate at which molecules were transferred to the surface from the near-surface layer. Direct counting of single-molecule events, made under steady-state conditions at extremely low coverage, provided direct, model-independent measurements of this attachment rate, in contrast with conventional ensemble-averaged methods, which are influenced by bulk transport and competing detachment processes. We found that the attachment rate increased with increasing temperature for all solvents. Arrhenius analyses gave activation energies of 5+/-2 kJ/mol for adsorption from HD, 10+/-2 kJ/mol for adsorption from THF, and 19+/-2 kJ/mol for adsorption from water. These energies increased systematically with the solvent polarity and, therefore, with the expected strength of the solvent-substrate interaction. We hypothesize that the adsorption of amphiphilic solute molecules from solution can be regarded as a competitive exchange between solute molecules and surface-bound solvent. In this scenario, adsorption is an activated process, and the activation energy for attachment is associated with the solvent-substrate interaction energy. PMID- 19563227 TI - Phase separation of palmitic acid and perfluorooctadecanoic acid in mixed Langmuir-Blodgett monolayer films. AB - Deposition of mixtures of palmitic acid (C15H31COOH) and perfluorooctadecanoic acid (C17F35COOH) onto solid substrates gives rise to irregularly shaped, phase separated domains under a variety of deposition conditions. The morphology and chemical composition of these phase-separated domains have been investigated using a combination of surface pressure-area isotherms, atomic force microscopy, X-ray photoemission electron microscopy, and confocal fluorescence microscopy imaging. While domain morphology and composition in 2D phase-separated mixed monolayer systems can typically be rationalized in terms of an interplay between line tension and dipole-dipole repulsion effects, it was found that for this system additional kinetic factors, including domain growth rates and the rate of dissolution of the fatty acid component into the aqueous subphase, also play a major role in controlling film properties. The potential importance of these effects for the controlled patterning of solid substrates is discussed. PMID- 19563228 TI - Covalent immobilization of antibacterial furanones via photochemical activation of perfluorophenylazide. AB - N-(3-trimethoxysilylpropyl)-4-azido-2,3,5,6-tetrafluorobenzamide (PFPA-silane) was used as a photoactive cross-linker to immobilize antibacterial furanone molecules on silicon oxide surfaces. This immobilization strategy is useful, especially for substrates and molecules that lack reactive functional groups. To this end, cleaned wafers were initially incubated in solutions of different concentrations of PFPA-silane to form a monolayer presenting azido groups on the surface. The functionalized surfaces were then treated with a furanone solution followed by illumination with UV light and extensive rinsing with ethanol to remove noncovalently adhered molecules. In the presented study, we demonstrate the ability to control the surface density of the immobilized furanone molecules by adjusting the concentration of PFPA-silane solution used for surface functionalization using complementary surface analytical techniques. The fluorine in PFPA-silane and the bromine in furanone molecules were convenient markers for the XPS study. The ellipsometric layer thickness of the immobilized furanone molecules on the surface decreased with decreasing PFPA-silane concentration, which correlated with a decline of water contact angle as a sign of film collapse. The intensity of characteristic azide vibration in the MTR IR spectra was monitored as a function of PFPA-silane concentration, and the peak disappeared completely after furanone application followed by UV irradiation. As a complementary technique to XPS, TOF-SIMS provided valuable information on the chemical and molecular structure of the modified surfaces and spatial distribution of the immobilized furanone molecules. Finally, this report presents a convenient, reproducible, and robust strategy to design antibacterial coating based on furanone compounds for applications in human health care. PMID- 19563229 TI - Variation of ion-exchange capacity, zeta potential, and ion-transport selectivities with the number of layers in a multilayer polyelectrolyte film. AB - The properties of poly(styrene sulfonate) (PSS)/poly(diallyldimethylammonium chloride) (PDADMAC) films vary dramatically with the number of polyelectrolyte layers deposited. Attenuated total reflectance infrared spectroscopy of (PDADMAC/PSS)n films deposited on a Ge crystal shows that coatings with fewer than 7 PDADMAC/PSS bilayers do not absorb significant amounts of SCN- or Ni(CN)4(2-) but coatings with more than 7 bilayers exhibit an ion-exchange capacity of about 0.5 mol/L of film. Consistent with ion-exchange, Ni(CN)4(2-) is the anion that is predominantly absorbed from equimolar mixtures of SCN- and Ni(CN)4(2-), even though SCN- initially exchanges into the film more rapidly than Ni(CN)4(2-). For silicon-supported PSS/PDADMAC films terminated with PSS, zeta potentials change from negative to positive as the number of adsorbed bilayers increases, presumably because of a high number of anion-exchange sites inside the film. These changes in film properties dramatically affect ion transport through (PSS/PDADMAC)nPSS-coated alumina membranes. The Cl-/SO4(2-) selectivities of these membranes are >30 with (PSS/PDADMAC)4PSS films but only 3 with (PSS/PDADMAC)6PSS films. Trends in zeta potentials and selectivities with increasing numbers of bilayers are consistent with the exponential growth mechanism, where a polycation absorbs throughout the film to create large numbers of anion-exchange sites, and during polyanion deposition, some of the polycation diffuses to the surface of the film to complex with polyanions from solution. Apparently, not all of the charge on the polycation is compensated by the polyanion; therefore, anion-exchange sites remain in the film, and the presence of this positive charge yields decreased Cl-/SO4(2-) selectivity. PMID- 19563230 TI - Phase behavior of an equimolar mixture of N-palmitoyl-D-erythro-sphingosine, cholesterol, and palmitic acid, a mixture with optimized hydrophobic matching. AB - The phase behavior and lipid mixing properties of an equimolar mixture of nonhydroxylated palmitoyl ceramide (Cer16), palmitic acid (PA), and cholesterol have been investigated using 2H NMR and vibrational spectroscopy. This mixture is formed by the three main classes of lipids found in the stratum corneum (SC), the top layer of the epidermis, and provides an optimized hydrophobic matching. Therefore, its behavior highlights the role played by hydrophobic matching on the phase behavior of SC lipids. We found that, below 45 degrees C, the mixture is essentially formed of coexisting crystalline domains with a small fraction of lipids (less than 20%) that forms a gel or fluid phase, likely ensuring cohesion between the solid domains. Upon heating, there is the formation of a liquid ordered phase mainly composed of PA and cholesterol, including a small fraction of Cer16. This finding is particularly highlighted by correlation vibrational microspectroscopy that indicates that domains enriched in cholesterol and PA include more disordered Cer16 than those found in the Cer16-rich domains. Solubilization of Cer16 in the fluid phase occurs progressively upon further heating, and this leads to the formation of a nonlamellar self-assembly where the motions are isotropic on the NMR time scale. It is found that the miscibility of Cer16 with cholesterol and PA is more limited than the one previously observed for ceramide III extracted from bovine brain, which is heterogeneous in chain composition and includes, in addition to Cer16, analogous ceramide with longer alkyl chains that are not hydrophobically matched with cholesterol and PA. Therefore, it is inferred that, in SC, the chain heterogeneity is a stronger criteria for lipid miscibility than chain hydrophobic matching. PMID- 19563231 TI - Fluorescence labeling and quantification of oxygen-containing functionalities on the surface of single-walled carbon nanotubes. AB - Fluorescence labeling of surface species (FLOSS) was applied to identify and determine the concentration of oxygen-containing functionalities on single-walled carbon nanotubes (SWCNTs), subjected to two different purification processes (air/HCl and nitric acid treatments) and compared to as-received (nonpurified) SWCNTs. The fluorophores were selected for their ability to covalently bind, with high specificity, to specific types of functionalities (OH, COOH, and CHO). FLOSS revealed that even as-received SWCNTs are not pristine and contain approximately 0.6 atomic % oxygen functionalities. FLOSS showed that, after nitric acid treatment, SWCNTs are approximately 5 times more functionalized than SWCNTs after air/HCl purification (5 versus 1 atomic % oxygen functionalities), supporting the idea that the former purification process is more aggressive than the latter. FLOSS demonstrated that carbonyls are the major functionalities on nitric-acid purified SWCNTs, suggesting that chemical derivatization strategies might consider exploiting aldehyde or ketone chemistry. PMID- 19563232 TI - Facile fabrication method and characterization of hollow Ag/SiO2 double-shelled spheres. AB - Hollow Ag/SiO2 double-shelled spheres with silver nanoparticle coating onto the interior wall of hollow silica have been successfully prepared via a novel and facile approach. In this method, negatively charged polystyrene (PS) beads were first modified by Sn2+ ions via electrostatic interaction, and then by Ag+ ions, which were reduced by Sn2+ ions and sodium-potassium tartrate to obtain PS/Ag composite spheres. When these spheres were coated by silica nanoparticles as shells from the hydrolysis and condensation reaction of tetraethoxysilane (TEOS) in isopropanol/ammonia mixture at 15 degrees C, the PS beads were "dissolved" in the same medium subsequently even synchronously, directly forming hollow Ag/SiO2 double-shelled spheres. Neither additional dissolution nor calcination process was used to remove polymer templates. This structure of hollow spheres not only has high catalytic activity, but also will decrease the loss of Ag nanoparticles due to frictional and/or other mechanical forces and the possible aggregations, and have controllable and selective catalytic activity compared to these common nanocomposite spheres with noble metal nanoparticles coating on the surfaces of supporting beads. PMID- 19563233 TI - Fabrication of metallic microtubes using self-rolled polymer tubes as templates. AB - We present a novel approach for fabricating single and bimetallic (gold, titanium) (Au, Ti, Au/Ti) microtubes with very high aspect ratio from self-rolled polymer templates. The polymer microtubes used as the template were generated by self-rolling of thin polymer bilayer films (polystyrene/poly(4-vinylpyridine) (PS/P4VP) gradually released from a solid substrate. The self-rolling was introduced in the polymer bilayer by swelling the bottom P4VP layer in dodecylbenzenesulfonic acid (DBSA) solution, which was opposed by a stiff top PS layer. The inner wall of the tube was metallized by depositing a thin layer of desired metal on top of the bilayer by physical vapor deposition. The polymer template was then removed by pyrolysis, resulting in pure metal microtubes, which were characterized by optical microscopy, scanning electron microscopy (SEM), infrared spectroscopy (IR), energy-dispersive X-ray (EDX), and X-ray photoelectron spectroscopy (XPS). The tube diameter was tailored by changing the metal layer thickness on the polymer bilayer. The approach described here is general and could be used to fabricate any type of single or multimetallic tube. The metal microtubes reported in this method have potential application in drug delivery systems, microelectronics, microfluidic devices, enzyme bireaction, and chemical and biochemical sensing devices. PMID- 19563234 TI - Interfacing neurons both extracellularly and intracellularly using carbon nanotube probes with long-term endurance. AB - This study demonstrates that carbon nanotubes (CNTs) can be fabricated into probes directly, with which neural activity can be monitored and elicited not only extracellularly but also intracellularly. Two types of CNT probes have been made and examined with the escape neural circuit of crayfish, Procambarus clarkia. The CNT probes are demonstrated to have comparable performance to conventional Ag/AgCl (silver/silver cloride) electrodes. Impedance measurement and cyclic voltammetry further indicate that the CNT probes transmit electrical signals through not only capacitive coupling but also resistive conduction. The resistive conduction facilitates the recording of postsynaptic potentials and equilibrium membrane potentials intracellularly as well as the delivery of direct current stimulation. Furthermore, delivering current stimuli for a long term is found to enhance rather than to degrade the recording capability of the CNT probes. The mechanism of this fruitful result is carefully investigated and discussed. Therefore, our findings here support the suggestion that CNTs are suitable for making biocompatible, durable neural probes of various configurations for diverse applications. PMID- 19563235 TI - Quantification of phytoestrogens in industrial waste streams. AB - Chemical compounds, including plant-based phytoestrogens, can function as hormone mimics and alter endocrine signaling in wildlife. In the present study, the waste streams from 19 plant-processing industries, including biofuel manufacturers, were sampled and analyzed for the phytoestrogens genistein, daidzein, coumestrol, formononetin, biochanin A, and zearalenone, via liquid chromatography/mass spectrometry. Eight of these industries contained phytoestrogens at environmentally relevant levels (>=1,000 ng/L), with the highest at approximately 250,000 ng/L. The influent and effluent streams of three municipal wastewater treatment plants receiving flow from some of these industries also were sampled and analyzed for the same phytoestrogens. It appeared that aerobic biological treatment, such as activated sludge, was able to remove these compounds from the liquid stream. Nevertheless, the effluent stream from one of the wastewater treatment plants had a phytoestrogen concentration above 1,000 ng/L. Results of the present study indicate the need for caution when designing facilities to treat the effluents from biofuel and other plant-processing industries. PMID- 19563236 TI - Pol bootscanning analysis of HIV type 1 can exhibit unexpected recombinations. AB - In France the recommendation is to sequence the RT gene of HIV-1 isolates prior to initiation of antiretroviral therapy. The data are routinely used for molecular characterization of the viruses yielding the subtype or CRF of the isolates investigated together with the absence or presence of drug resistance mutations. In this study, we performed bootscanning analysis on the whole pol gene, in which in vitro and in vivo intersubtype recombination has been reported to occur frequently. We showed that out of 15 HIV-1 isolates, two exhibited a recombination unexpected by this routine sequencing method. PMID- 19563237 TI - Cases of HIV type 1 acute infection at STI-related clinics in Osaka. AB - Since 1992 we have investigated HIV antibodies in persons with a venereal disease who worked for a sex business or engaged in sexual behavior with a high risk of HIV infection at five clinics of venereology, urology, and gynecology in Osaka Prefecture. Starting at the end of 2000, we performed a nucleic acid amplification test (NAT) on antibody-negative samples with the aim of detecting cases in the window period covering the early phase of infection. Three cases that were thought to be in this phase were found in 2006. All were confirmed to be positive in an NAT, although they were negative in initial screening for HIV performed using the IC method. Gene analysis of the env-C2V3 region of HIV-1 showed that the three samples had different origins. PMID- 19563238 TI - Modulation of K65R selection by zidovudine inclusion: analysis of HIV resistance selection in subjects with virologic failure receiving once-daily abacavir/lamivudine/zidovudine and tenofovir DF (study COL40263). AB - COL40263 was a pilot 48-week, open-label, multicenter study evaluating the efficacy and safety of once-daily coformulated abacavir/lamivudine/zidovudine plus tenofovir in ART-naive, HIV-infected subjects. We examined the patterns of resistance that were selected on-therapy through 48 weeks in subjects with virologic nonresponse (VF). A total of 123 antiretroviral-naive HIV-1-infected subjects with plasma HIV-1 RNA > or = 30,000copies/ml were enrolled. For subjects with confirmed VF (HIV-1 RNA > or = 400 copies/ml at week 24 or later), HIV population genotypic and phenotypic analysis was performed. Of the 123 enrolled subjects, 14 (11%) had confirmed plasma HIV-1 RNA > or = 400 copies/ml through week 48. Of these subjects, 3/14 had evidence of drug resistance at baseline: 2/14 had HIV with K103N, Y188F/H/L/Y, and/or T215A and 1/14 had reduced zidovudine susceptibility. At the last time point analyzed, 4/14 subjects had wild-type HIV, while 10/14 subjects had HIV with either thymidine analogue mutations (TAMS) alone (3/10), TAMS + M184V (4/10), M184V only (1/10), or K65R/K (2/10). Matched phenotype was obtained for 13/14 subjects and 8/13 (62%) subjects had reduced susceptibility to one or more study drugs: 2/13 tenofovir, 3/13 abacavir, 4/13 zidovudine, and 7/13 lamivudine. The resistance pattern in COL40263 subjects with VF differs significantly from that reported for tenofovir containing triple-nucleoside regimens. TAMs were detected in the majority (7/10) of samples from subjects with VF who selected any resistance mutation. These data suggest that TAMs selection is a preferred resistance route of this combination, with zidovudine modulating the resistance pathway against selection for K65R. PMID- 19563239 TI - Longitudinal study on the influence of Nd:YAG laser irradiation on microleakage associated with two filling techniques. AB - OBJECTIVE: This study investigates the effects of Nd:YAG laser irradiation on apical and coronal seals, when used prior to two root canal filling techniques. BACKGROUND DATA: Limited information exists regarding the effects of morphologic changes to dentin walls following Nd:YAG laser irradiation on the sealing ability of root fillings. METHODS: Two hundred forty teeth were analyzed by observing coronal and apical leakage of Indian ink (DL), and 60 were analyzed for through and-through leakage using the fluid transport model (FTM). The Nd:YAG laser parameters were 1.5 W, 100 mJ, and 15 Hz (four times for 5 s at 20 s intervals). Each group consisted of a lased and a nonlased subgroup: each subgroup had root fills done by either cold lateral condensation (CLC) or hybrid condensation (HC). Leakage was assessed after 48 h, and then at 1, 6, and 12 months. The DL group was divided into four groups of 15 teeth for each evaluation point. Through-and through leakage (L in microliters/day) was measured for 48 h under a pressure of 1.2 atm using FTM, and recorded as L = 0 (L1), 0 < L 10 (L3). RESULTS: Apical and coronal dye leakage was observed in all groups. Significant differences (p < 0.05) in apical leakage were found between HC and HC + Nd after 1, 6, and 12 months, and between CLC and CLC + Nd at 6 and 12 months. No significant differences were found between laser-irradiated and non-laser irradiated groups with FTM. CONCLUSION: Pulsed Nd:YAG laser irradiation following root canal preparation may reduce apical leakage in association with hybrid gutta percha condensation. PMID- 19563241 TI - Physical and compositional changes on demineralized primary enamel induced by CO2 Laser. AB - OBJECTIVE: This in vitro study aimed to evaluate the physical and chemical changes promoted by a CO(2) laser at 10.6-microm wavelength on primary dental enamel with artificial caries-like lesions. BACKGROUND DATA: Several previous investigations have shown that enamel can be modified by CO(2) laser to obtain a caries-preventive effect, but the specific mechanism remains uncertain. MATERIALS AND METHODS: Twenty-seven primary molars were randomly assigned to three groups as follows: control, carious, and laser (n = 9). The specimens from the carious and laser groups were demineralized and treated with or without CO(2) laser, according to the group. Enamel surface changes after treatments were monitored using Fourier transform Raman spectroscopy and scanning electron microscopy (SEM). RESULTS: The Raman spectra showed a statistically significant reduction of mineral content in carious and laser groups when compared to control group. Additionally, carbonate content was reduced in irradiated specimens when compared to the other groups. No physical change was observed in specimens evaluated by SEM. CONCLUSION: The results suggest that CO(2) laser irradiation may reduce the carbonate content of enamel, which is likely to make this substrate more acid resistant. PMID- 19563242 TI - Different applications of monochromatic excimer light in skin diseases. AB - BACKGROUND: Ultraviolet radiation has been used for curative purposes in dermatologic conditions, especially in the last 30 years. OBJECTIVES: We analyzed the efficacy of monochromatic excimer light in psoriasis, palmoplantar pustulosis, vitiligo, mycosis fungoides and alopecia areata, and to examine potential new indications. METHODS: Two hundred seventy-nine patients with common and persistent skin diseases were enrolled in an open prospective study: 152 patients with stable and localized plaque psoriasis, 47 with palmoplantar psoriasis, 7 with palmoplantar pustulosis, 32 with vitiligo, 11 with prurigo nodularis, 9 with mycosis fungoides stage Ia, 8 with alopecia, 5 with localized scleroderma, 5 with genital lichen sclerosus, and 3 with granuloma annulare. The 308 nm excimer light was used at a power density of 48 mW/cm(2). An average of 12 sessions (range, 6-18), one session per week, was performed and yielded a total dose range of 4-12.5 J/cm(2). Clinical response was assessed using photos, biopsies, and specific clinical scores. Patients were monitorized for 6 and 12 months for psoriasis, 12 months for mycosis fungoides, and 4 months for the remaining conditions. RESULTS: We observed complete remission in more than 50% of patients with plaque psoriasis and palmoplantar dermatoses, respectively, complete remission in all patients affected by mycosis fungoides, excellent repigmentation in one third of vitiligo patients, hair regrowth in three patients with alopecia areata, an overall improvement in prurigo nodularis, a partial remission in patients affected by localized scleroderma, and a complete remission in most of the patients with genital lichen sclerosus and granuloma annulare. CONCLUSIONS: Our study confirms the use of monochromatic excimer light as a valid choice for the treatment of psoriasis, vitiligo, and mycosis fungoides; we also observed and report for the first time that monochromatic excimer light produces a therapeutic response in prurigo nodularis, localized scleroderma, genital lichen sclerosus, and granuloma annulare. PMID- 19563243 TI - Radiant power determination of low-level laser therapy equipment and characterization of its clinical use procedures. AB - OBJECTIVE: The main objectives of this study were to characterize low-level laser therapy (LLLT) and the physical therapy clinical procedures for its use. BACKGROUND DATA: There are few scientific studies that characterize the calibration of LLLT equipment. MATERIALS AND METHODS: Forty lasers at 36 physical therapy clinics were selected. The equipment was characterized through data collected from the owner manuals, direct consultation with the manufacturers, and a questionnaire answered by the users. A digital potency analyzer was used to calibrate released mean potency. Qualitative data were presented throughout the descriptive statistics and quantitative data were analyzed by the Wilcoxon/Kruskal-Wallis and Fisher tests (significance, p < 0.05). RESULTS: The laser equipment was either AsGa (70.5%) or HeNe (23.5%), and 60% was analog and acquired over 5 years ago. The majority of the equipment was used 10-15 times per week and the most frequent density level used was 2 to 4 J/cm(2). Protective goggles were available in only 19.4% of the clinics evaluated. The association between the analyzed categories demonstrated that a lower mean potency was correlated both with equipment acquired over 5 years ago and analog technology. The determined mean potency was lower than the one claimed by the manufacturer (p < 0.05). In 30 cases, the analyzed equipment presented a potency between 3 microW and 5.6 mW; in three cases, the potency was >25 mW; and in seven cases, potency was nonexistent. CONCLUSION: The analyzed equipment was out-dated and periodical maintenance was not conducted, which was reflected in the low irradiated potency. PMID- 19563244 TI - High pregnancy-related anxiety and prenatal depressive symptoms as predictors of intention to breastfeed and breastfeeding initiation. AB - BACKGROUND: Perinatal mood disorders affect up to 20% of women in the United States. Little is known about how disorders in maternal mood may affect rates of breastfeeding. OBJECTIVE: To determine the impact of prenatal depressive symptoms and high pregnancy-related anxiety on (1) prenatal intention to breastfeed and (2) breastfeeding initiation. METHODS: We prospectively followed 1436 pregnant women enrolled in the cohort study Project Viva. The main outcome measures were (1) mother's second trimester self-report of intention to use all or mostly formula in the first week of life and (2) failure to initiate breastfeeding. We defined prenatal depressive symptoms as a second trimester Edinburgh Postpartum Depression Scale (EPDS) score of > or =13 and high pregnancy-related anxiety as a "very much" response to three or more questions on a first trimester pregnancy anxiety scale. RESULTS: Of the 1436 participants, 9% (n = 125) had prenatal depressive symptoms indicative of depression, and 10% (n = 141) reported high pregnancy-related anxiety; 11% (n = 159) intended to give mostly or only formula in the first week of life, and 86% (n = 1242) initiated breastfeeding. In multivariate analyses, women with prenatal depressive symptoms (OR 1.92, 95% CI 1.11, 3.33) and high pregnancy-related anxiety (OR 1.99, 95% CI 1.12, 3.54) were roughly two times more likely than women without these mood disorders to plan to formula feed. However, neither prenatal depressive symptoms (OR 1.06, 95% CI 0.61, 1.84) nor high pregnancy-related anxiety (OR 1.28, 95% CI 0.74, 2.20) was associated with failure to initiate breastfeeding. CONCLUSIONS: In a healthcare setting highly supportive of breastfeeding, women with prenatal depressive symptoms and possibly those with high pregnancy-related anxiety were less likely to plan prenatally to breastfeed, although this tendency did not translate into lower breastfeeding initiation rates. PMID- 19563245 TI - Patient satisfaction in postmenopausal women treated with a weekly bisphosphonate transitioned to once-monthly ibandronate. AB - OBJECTIVE: CURRENT, a large, open-label, 6-month, multicenter study, was designed to assess patient satisfaction levels and patient treatment preference after switching from weekly oral bisphosphonates to monthly oral ibandronate for a period of 6 months. METHODS: This study enrolled postmenopausal women who had taken a weekly oral bisphosphonate for at least 3 months for prevention or treatment of osteoporosis or osteopenia at the time of screening. Enrolled patients were switched to 150 mg monthly ibandronate. At baseline and 6 months, patients completed the Osteoporosis Patient Satisfaction Questionnaire (OPSAT-Q), consisting of four domains. Scores were converted to composite satisfaction scores (scale of 0-100). At 6 months, patients completed the Preference Questionnaire. Adverse events were monitored throughout. RESULTS: The intent-to treat population comprised 1678 patients. OPSAT-Q composite satisfaction scores improved by 9 points by month 6 despite the high mean baseline summary scores (80.1 points). Convenience, overall satisfaction, and quality of life domain scores improved by 15.6, 12, and 9.2 points, respectively. Increased satisfaction was reported by the majority of patients at month 6 (70.4%). Patients who reported stomach upset or suboptimal compliance with prestudy weekly bisphosphonate treatment were more likely to report improved satisfaction (odds ratio [OR] for stomach upset 2.98, 95% CI 1.52, 6.50, p = 0.0026; suboptimal compliance 1.82, 95% CI 1.13-3.04, p = 0.017). After 6 months, 73.6% of patients preferred monthly ibandronate to weekly bisphosphonates. The most frequently occurring adverse events were upper respiratory tract infection (3.2% of patients), dyspepsia (2.5%), fracture (2.4%), arthralgia (2.3%), and gastroesophageal reflux disease, diarrhea, and nausea (2.2% each). CONCLUSIONS: Patients previously using weekly bisphosphonates reported improved satisfaction with monthly ibandronate dosing. PMID- 19563246 TI - Comparing the health burden of eating-disordered behavior and overweight in women. AB - OBJECTIVE AND METHODS: We compared the health burden of eating-disordered behavior with that of overweight in a community-based sample of women aged 18-42 years residing in the Australian Capital Territory region of Australia. Participants (n = 4643) completed self-report measures of eating disorder psychopathology, health-related quality of life and health service utilization. Body mass index (BMI) was derived from self-reported height and weight. RESULTS: Overweight was associated with marked impairment in physical health functioning and comparatively little impairment in psychosocial functioning, whereas eating disordered behavior was associated with marked impairment in psychosocial functioning and comparatively little impairment in physical health functioning. Further, (1) impairment in psychosocial functioning associated with eating disordered behavior was greater than impairment in physical health functioning associated with overweight, and (2) impairment in physical health functioning associated with eating-disordered behavior was greater than impairment in psychosocial functioning associated with overweight. Overweight and eating disordered behavior were associated with similarly elevated rates of primary care consultations during the past 6 months and of lifetime treatment from a health professional for an eating or weight problem. CONCLUSIONS: In young adult women, the health burden of eating-disordered behavior may be more substantial than previously recognized. Better information concerning the spectrum of disordered eating that exists at the population level needs to be made available. Eating disordered behavior warrants greater attention when considering the public health burden of obesity and in developing programs to reduce this burden. PMID- 19563247 TI - Toward optimal health: advising aging women about dementia. Interview by Jodi R. Godfrey. PMID- 19563248 TI - Women with safety concerns report fewer gender-specific preventive healthcare services. AB - BACKGROUND: Violence and safety exposures, primarily in the form of intimate partner violence (IPV) exposures, have been associated with increased healthcare utilization in women and variable associations with preventive services. It remains unknown, however, if broader measures of violence and safety exposures are associated with preventive healthcare utilization. METHODS: Data from the 2004 The Philadelphia Health Management Corporation survey were used to compare self-reported utilization of preventive healthcare services for participants who did and did not report exposures to violence or have safety concerns during the past year, using novel but nonvalidated measures of violence and safety concerns. The sample included women aged 18-96 years who reported having a usual source of care. Measures of preventive healthcare utilization included timing of last mammogram, cervical cytology, breast examination, blood cholesterol testing, blood pressure measurement, and screening for colon cancer. RESULTS: The survey had an American Association for Public Opinion Research response rate of 30.4%. Of the 6285 women surveyed who reported having a usual source of care, 456 (7.3%) reported either an exposure to violence or safety concerns, and 5821 did not. Using logistic regression models, women aged 18-65 years who reported safety concerns were less likely to report having cervical cancer testing in the past 12 months (OR 0.68, 95% CI 0.49-0.94). Women > or =40 years who reported safety concerns in the last year were less likely to report having clinical breast examinations (OR 0.63, 95% CI 0.43-0.93) or mammography (OR 0.57, 95% CI 0.39 0.83). Physical violence exposure was not significantly associated with preventive healthcare utilization. CONCLUSIONS: In this large population-based sample, women with recent self-reported safety concerns report receiving fewer recommended gender-specific preventive services. PMID- 19563249 TI - No association between vitamin D receptor polymorphisms and coronary artery disease in a Chinese population. AB - The clinical features suggest that genetic factors may have a strong influence on susceptibility to coronary artery disease (CAD). The aim of this study was to investigate the association between FokI (rs2228570) and BsmI (rs1544410) of the vitamin D receptor (VDR) gene polymorphisms and patients with CAD in a Chinese population. One hundred and fifty-two CAD patients and 212 healthy controls were genotyped for the FokI and BsmI polymorphisms in VDR gene using polymerase chain reaction-restriction fragment length polymorphism. No significant differences were observed in the genotype and allele frequencies of the FokI and BsmI polymorphisms between the cases and controls (For FokI: odds ratio = 1.11, 95% confidence interval 0.83-1.50; for BsmI: odds ratio = 0.74, 95% confidence interval 0.44-1.23). There was no significant difference in the genotype distribution or the allele frequencies of VDR FokI and BsmI between two groups in a Chinese population. PMID- 19563250 TI - Molecular genetic relationship of the 3' untranslated region among Thai dengue-3 virus, Bangkok isolates, during 1973-2000. AB - Dengue virus serotype 3 (DENV-3) was associated with severe dengue epidemics in Thailand during 1973-1999. We studied Thai DENV-3 viruses isolated from hospitalized children in Bangkok with differing disease severity during that period. Viruses were sequenced at their 5' and 3' untranslated regions (UTRs), which are regions that play a pivotal role in viral replication. Our results indicated that the primary sequences as well as the secondary structures at both ends of Thai DENV-3 viruses were highly conserved over almost three decades. We found nucleotide insertions and deletions at the variable region (VR) that is located just downstream of the nonstructural protein 5 (NS5) stop codon among these viruses. The phylogenetic tree derived from the size heterogeneity of VR in the 3' UTR divided DENV-3 into four genotypes, and Thai DENV-3 viruses in this study belonged to genotype II. The replication efficiency of the candidate viruses with different lengths at the VR were assessed in the mosquito (C6/36) and human (HepG2) cell lines. Our results show that the viruses with nucleotide insertions at VR replicated better than the virus that contained deletions. Our findings indicate that Thai DENV-3 demonstrated a remarkable conservation of nucleotides over 28 years. Correlation with disease severity suggests that both primary sequences and secondary structures of the 3' UTR do not appear correlated with disease severity in humans. PMID- 19563251 TI - In vitro study of damaging effects of 2,4-dichlorophenoxyacetic acid on DNA structure by spectroscopic and voltammetric techniques. AB - 2,4-Dichlorophenoxyacetic acid (2,4-D) is a genotoxic organochlorinated herbicide, and its interaction with DNA was studied by UV/Vis, fluorescence, circular dichroism viscosity measurements, and alternative current voltammetry techniques. Using these analyses, the binding constant of 2,4-D to DNA has been calculated by two different techniques. The binding constant of 2,4-D to DNA calculated by fluorescence and circular dichroism spectra was found to be 3.5 x 10(3) M(-1) and 5.02 x 10(3) M(-1), respectively. Analyses of fluorescence spectra, viscosity measurements, and alternative current voltammetry interactions indicated that 2,4-D is a groove binder of DNA. Ethidium bromide displacement studies revealed that 2,4-D does not have any effect on ethidium bromide-bound DNA, which is indicative of groove binding. PMID- 19563252 TI - In vitro study of calf thymus DNA interaction with butylated hydroxyanisole. AB - The interaction of native calf thymus DNA with butylated hydroxyanisole (BHA) at physiological pH was investigated by spectrofluorometric, voltammetric, and viscosimetric techniques. BHA molecules intercalated between base pairs of DNA, demonstrated by a sharp increase in specific viscosity of DNA and decreases in the fluorescence of BHA solutions in the presence of increasing amounts of DNA and calculated formation constant (K(f)) at different temperatures. Further, the enthalpy and entropy of the reaction between BHA and calf thymus DNA showed that the reaction is exothermic and enthalpy favored (DeltaH = -89.02 kJ/mol; DeltaS = -211.21 J/mol K). Also, peak current decrease and positive shift in the cyclic voltammetry indicate that BHA is able to intercalate in the DNA base pairs. PMID- 19563253 TI - A new sterilization technique of bovine pericardial biomaterial using microwave radiation. AB - Bioprosthetic valves created from chemically treated natural tissues such as bovine pericardial biomaterial are used as heart valve scaffolds. Methods currently available for sterilization of biomaterial for transplantation include the application of gamma radiation and chemical sterilants. These techniques, however, can be problematic because they can be expensive and lead to a reduction in tissue integrity. Therefore, improved techniques are needed that are cost effective and do not disrupt the physical properties, functionality, and lifespan of the valvular leaflets. This study examined a novel technique using nonthermal microwave radiation that could lead to the inactivation of bacteria in bovine pericardial biomaterial without compromising valve durability. Two common pathogenic species of bacteria, Escherichia coli and Staphylococcus aureus, were used as test microorganisms. Optimized microwave parameters were used to determine whether inactivation of pathogenic bacteria from bovine pericardium could be achieved. In addition, the effect of microwave sterilization on tissue integrity was examined. The mechanical properties (assessed using dynamic mechanical analysis) and tensile strength testing (using a Universal Tensile Tester) as well as thermal analysis (using thermogravimetric analysis and differential scanning calorimetry) indicated that microwave sterilization did not compromise the functionality of bovine pericardial biomaterial. Scanning electron microscopy imaging and cytotoxicity testing also confirmed that the structure and biocompatibility of transplant biomaterial remained unaltered after the sterilization process. Results from the application of this new microwave (MW) sterilization technique to bovine pericardium showed that near-complete inactivation of the contaminant bacteria was achieved. It is concluded that nonthermal inactivation of pathogenic bacteria from bovine pericardial biomaterial could be achieved using microwave radiation. PMID- 19563254 TI - Spontaneous formation of highly functional three-dimensional multilayer from human hepatoma Hep G2 cells cultured on an oxygen-permeable polydimethylsiloxane membrane. AB - When considering high-density liver cell cultures, adequate delivery of oxygen to the cells appears particularly crucial since it is one of the most limiting parameters of hepatocytes' functions. Here we report on the effects of direct oxygenation through a gas-permeable polydimethylsiloxane membrane on liver derived cell culture. We used highly proliferative human hepatoma Hep G2 cells to assess the growth-related limitation of such direct oxygen supply. As a consequence of the greater oxygen availability, the proliferation of Hep G2 cells was markedly enhanced, leading to the formation of a thick three-dimensional cellular multilayer. This was supported by the oxygen concentration profiles in the vicinity of the cell layers, predicted by numerical simulations. The applied cells also displayed an increased functionality as assessed by the high-albumin production rates, with per well-based albumin secretion reaching after 15 days 50 times that of the cells cultured in conventional polystyrene plates. Anaerobic glycolysis was in the same time significantly reduced as assessed by both the reduced glucose consumption and lactate production rates. Direct oxygenation through such a gas-permeable membrane appears suitable for engineering functional thick liver tissues for in vivo implantation, as well as for microplate-based chemical/drug screening applications. PMID- 19563255 TI - Online monitoring of the mechanical behavior of collagen hydrogels: influence of corneal fibroblasts on elastic modulus. AB - Collagen hydrogels have been widely used to model biological systems and examine cell behavior in vitro. Of increasing interest is how cells affect the mechanical characteristics of their surrounding matrix and vice versa over long culture periods. In this study, the change in mechanical properties of collagen hydrogels embedded with human corneal fibroblasts was examined over a 6-week culture period using a novel online spherical indentation system. The elastic modulus of the hydrogels was found to increase during the first 2 weeks of culture and decrease after 4 weeks in culture. The effects of actin polymerization and matrix metalloproteinase inhibitors were also examined, which verified some mechanisms involved in the alteration of the mechanical properties such as cell tensile forces and other potential factors. This online monitoring technique demonstrates the ability to examine the mechanical properties of cell-seeded constructs in response to the culture environments--in particular, the response to the addition of drugs or chemical reagents--which will provide a useful tool in studying the mechano-feedback loop between cells and their surrounding matrix. PMID- 19563256 TI - Cost-effectiveness of ziconotide in intrathecal pain management for severe chronic pain patients in the UK. AB - OBJECTIVE: To examine the cost-effectiveness of using intrathecal ziconotide in the treatment of severe chronic pain compared to best supportive care for patients with intractable chronic pain in the United Kingdom. METHODS: Using a simulation model, the analysis evaluated the cost and health economic consequences of using ziconotide as a treatment for severe chronic pain. The modelled population and clinical data were based on a randomised controlled trial in which the main outcome was reduction in pain as measured by the visual analogue scale of pain intensity (VASPI). Resource use data were elicited using a modified Delphi panel and costed using published sources. Utility values were derived from a separate research study. The main outcome measure was the cost per quality-adjusted life-year (QALY). Extensive scenario analysis was conducted to evaluate parameter uncertainty. RESULTS: Overall, findings were robust to most assumptions. The cost-effectiveness of ziconotide compared to best supportive care (BSC) was pound 27,443 per QALY (95% CI pound 18,304-38,504). Scenarios were investigated in which discount rates, the time horizon, the threshold for qualifying as a responder, pump-related assumptions, utilities, ziconotide drug dose, and the patient discontinuation rate with ziconotide were varied. The most sensitive parameter was the dosage of ziconotide: using the lower and upper bounds of the average ziconotide dosage observed in the long-term open-label study changed the incremental cost-effectiveness ratio (ICER) to pound 15,500 [pound 8206-25,405] and pound 44,700 [pound 30,541-62, 670]. CONCLUSIONS: Ziconotide may offer an economically feasible alternative solution for patients for whom current treatment is inappropriate or ineffective. The main study limitation is that some model inputs, mainly related to resource use, are based on assumptions or expert interviews. PMID- 19563258 TI - Altered proinflammatory cytokine production and enhanced resistance to Trypanosoma congolense infection in lymphotoxin beta-deficient mice. AB - BALB/c mice are highly susceptible to Trypanosoma congolense infection, whereas C57BL/6 mice are relatively resistant. Overproduction of interferon-gamma (IFN gamma) and other proinflammatory cytokines contribute to death in susceptible mice. Here, we show that lymphotoxin beta-deficient (LTbeta(-/-)) mice are more resistant than wild-type (WT) mice to T. congolense infection, as shown by a lower parasitemia level and a longer survival duration. The enhanced resistance of LTbeta(-/-) mice was associated with undetectable or low serum levels of proinflammatory cytokines (i.e., tumor necrosis factor-alpha, interleukin [IL]-6, IL-12, and monocyte chemotactic protein-1). Although infected LTbeta(-/-) mice had high numbers of CD4(+)CD25(+)Foxp3(+) cells and high serum IL-10 levels, these cells were not the major producers of IL-10. Treatment of LTbeta(-/-) mice with anti-IL-10R monoclonal antibody abolished their enhanced resistance, whereas depletion of CD25(+) cells further enhanced resistance among infected WT and LTbeta(-/-) mice. These results suggest that LTbeta plays critical role in regulating the outcome of T. congolense infection in mice. PMID- 19563257 TI - Ciprofloxacin dosage and emergence of resistance in human commensal bacteria. AB - BACKGROUND: Although optimization of the fluoroquinolone dosage increases the efficacy of this class of drugs against bacterial infections, its impact on the emergence of resistance in commensal bacteria is unknown. METHODS: Six different 14-day dosages of oral ciprofloxacin were randomly assigned to 48 healthy volunteers. Individual pharmacokinetic and pharmacodynamic parameters combining antibiotic exposure in plasma, saliva, and stool specimens and ciprofloxacin minimum inhibitory concentrations (MICs) and mutant prevention concentrations against viridans group streptococci in the pharyngeal flora and Escherichia coli in the fecal flora were estimated. Their links with the emergence of resistance to nalidixic acid or ciprofloxacin in the fecal flora and to levofloxacin in the pharyngeal flora 7, 14, or 42 days after ciprofloxacin initiation were investigated. RESULTS: Resistance emerged in the fecal and pharyngeal flora of 25% and 33% of the subjects, respectively, mainly when local concentrations of ciprofloxacin were less than the MIC. No variable that integrated pharmacokinetic data and pharmacodynamic parameters was found to differ significantly between the subjects in whom resistance emerged and those in whom it did not. Probabilities of the emergence of resistance were not significantly different across the different antibiotic dosages. CONCLUSIONS: Selection of resistant commensals during ciprofloxacin therapy is a frequent ecological side effect that is not preventable by dosage optimization. Trial registration. Clinical Trials.gov identifier: NCT00190151. PMID- 19563259 TI - Assessment of the intimal response to a protein-modified stent in a tissue engineered blood vessel mimic. AB - Protein-coated intravascular stents have emerged as potential pro-healing modifications for or alternatives to anti-proliferative drug-eluting stents. To support the development of these devices, preclinical testing is required to evaluate the intimal response to new coatings and modifications. The purpose of this work was to implement a tissue-engineered blood vessel as an in vitro testing system to evaluate extracellular matrix-modified stents with regard to endothelialization of the stent surface. Stents were modified by submersion in a protein-enriched medium and were subsequently deployed within tissue-engineered blood vessels and cultivated in vitro under flow to assess the intimal response. Scanning electron microscopy, fluorescent nuclear staining with en face imaging, and histological assessments were performed 7 or 14 days postdeployment. Results illustrated accelerated cellular regeneration over protein-modified stent strut surfaces, with increased coverage and increased tissue thickness atop protein modified stent struts. In addition, the intimal response to modified stents differed significantly from bare metal stents. Conclusions from this work support the use of a tissue-engineered blood vessel mimic system for evaluation of modified stent surfaces. These findings are important to stent researchers as well as laboratories developing tissue-engineered constructs. PMID- 19563260 TI - Effect of the alphaGal epitope on the response to small intestinal submucosa extracellular matrix in a nonhuman primate model. AB - The Galalpha1,3Galbeta1,4GlcNAc-R (Gal) epitope is a major factor in the hyperacute rejection of pig organ transplants in primates. Biologic scaffold materials used for tissue reconstruction and composed of xenogeneic extracellular matrix (ECM) may contain the Gal epitope. However, the effect of this epitope upon the host response is controversial. The present study investigated the effect of the Gal epitope upon the host response to a porcine-derived ECM in an African Green monkey (Cholrocaebus aethiops) abdominal wall resection model. Histologic methods, serology, complement-dependent cytotoxicity, and gene expression profiling were used to evaluate the host response to allogeneic and both wild-type and Gal-deficient xenogeneic scaffold materials. Although expression of the Gal epitope induced an increase in serum anti-Gal antibodies in recipients, no other differences were noted in the host response between test articles. All ECM scaffolds were well tolerated and showed constructive remodeling during the study period. Recipients of all test articles showed no histologic or humoral evidence of sensitization when a second scaffold was implanted 45 days after the original surgery. The findings of the present study show that the presence of the Gal epitope within a porcine-derived ECM scaffold material elicits a serum antibody response, but no adverse effect upon tissue remodeling. PMID- 19563261 TI - Vascular smooth muscle cells as a valvular interstitial cell surrogate in heart valve tissue engineering. AB - BACKGROUND: Vascular smooth muscle cells (VSMCs) are a potential autologous cell source for aortic valve tissue engineering, but have a phenotype that differs from that of valvular interstitial cells in vivo. We hypothesized that combining basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), or platelet derived growth factor (PDGF) with transforming growth factor beta-1 (TGF-beta1) would achieve a valvular interstitial cell-like phenotype of VSMCs. METHODS: VSMC phenotype was assessed by immunofluorescence, proliferation was measured by the tetrazolium reduction (MTT) assay, and extracellular matrix gene expression was determined by real-time polymerase chain reaction. RESULTS: Combinations of growth factors that included PDGF showed the greatest increases in proliferation. Immunofluorescence for alpha-smooth muscle actin demonstrated an inverse correlation between proliferation and a myofibroblast-like phenotype, while combinations of TGF-beta1+ EGF+bFGF (TEF) and TGF-beta1+EGF+PDGF (TEP) induced the greatest change of alpha-smooth muscle actin expression compared to untreated controls. Finally, TEP treatment showed an increase in versican, fibronectin, and type I collagen mRNA expression, while decreasing matrix metalloproteinase 1 expression. CONCLUSIONS: Combination of TGF-beta1 with EGF and PDGF induces VSMC proliferation and expression of extracellular matrix constituents found in the aortic valve. In vitro preconditioning of VSMCs provides a potentially viable surrogate cell source for developing a valve graft. PMID- 19563262 TI - "Prepandemic" immunization for novel influenza viruses, "swine flu" vaccine, Guillain-Barre syndrome, and the detection of rare severe adverse events. AB - The availability of immunogenic, licensed H5N1 vaccines and the anticipated development of vaccines against "swine" influenza A(H1N1) have stimulated debate about the possible use of these vaccines for protection of those exposed to potential pandemic influenza viruses and for immunization or "priming" of populations in the so-called "prepandemic" (interpandemic) era. However, the safety of such vaccines is a critical issue in policy development for wide-scale application of vaccines in the interpandemic period. For example, wide-scale interpandemic use of H5N1 vaccines could lead to millions of persons receiving vaccines of uncertain efficacy potentially associated with rare severe adverse events and against a virus that may not cause a pandemic. Here, we first review aspects of the 1976 National Influenza Immunization Programme against "swine flu" and its well-documented association with Guillain-Barre syndrome as a case study illustration of a suspected vaccine-associated severe adverse event in a mass interpandemic immunization setting. This case study is especially timely, given the recent spread of a novel influenza A(H1N1) virus in humans in Mexico and beyond. Following this, we examine available safety data from clinical trials of H5N1 vaccines and briefly discuss how vaccine safety could be monitored in a postmarketing surveillance setting. PMID- 19563264 TI - Mitochondria: determinants of stem cell fate? AB - Stem cells are traditionally classified as being either embryonic stem cells (ESCs) or somatic stem cells. Such a designation has now become blurred by the advent of ostensibly pluripotent cells derived from somatic cells, referred to as induced pluripotent stem cells. Mitochondria are the membrane bound organelles that provide the majority of a cell's chemical energy via their production of adenosine triphosphate. Mitochondria are also known to be vital components in many cell processes including differentiation and apoptosis. We are still remarkably uninformed of how mitochondrial function affects stem cell behavior. Reviewed evidence suggests that mitochondrial function and integrity affect stem cell viability, proliferative and differential potential, and lifespan. Mitochondrial status therefore has profound and as yet unexamined implications for the current drive to develop induced pluripotent stem cells as a therapeutic resource. PMID- 19563263 TI - Tissue formation and vascularization in anatomically shaped human joint condyle ectopically in vivo. AB - Scale-up of bioengineered grafts toward clinical applications is a challenge in regenerative medicine. Here, we report tissue formation and vascularization of anatomically shaped human tibial condyles ectopically with a dimension of 20 x 15 x 15 mm(3). A composite of poly-epsilon-caprolactone and hydroxyapatite was fabricated using layer deposition of three-dimensional interlaid strands with interconnecting microchannels (400 microm) and seeded with human bone marrow stem cells (hMSCs) with or without osteogenic differentiation. An overlaying layer (1 mm deep) of poly(ethylene glycol)-based hydrogel encapsulating hMSCs or hMSC derived chondrocytes was molded into anatomic shape and anchored into microchannels by gel infusion. After 6 weeks of subcutaneous implantation in athymic rats, hMSCs generated not only significantly more blood vessels, but also significantly larger-diameter vessels than hMSC-derived osteoblasts, although hMSC-derived osteoblasts yielded mineralized tissue in microchannels. Chondrocytes in safranin-O-positive glycosaminoglycan matrix were present in the cartilage layer seeded with hMSC-derived chondrogenic cells, although significantly more cells were present in the cartilage layer seeded with hMSCs than hMSC-derived chondrocytes. Together, MSCs elaborate substantially more angiogenesis, whereas their progenies yield corresponding differentiated tissue phenotypes. Scale up is probable by incorporating a combination of stem cells and their progenies in repeating modules of internal microchannels. PMID- 19563265 TI - Inactivation of animal and human prions by hydrogen peroxide gas plasma sterilization. AB - Prions cause various transmissible spongiform encephalopathies. They are highly resistant to the chemical and physical decontamination and sterilization procedures routinely used in healthcare facilities. The decontamination procedures recommended for the inactivation of prions are often incompatible with the materials used in medical devices. In this study, we evaluated the use of low temperature hydrogen peroxide gas plasma sterilization systems and other instrument-processing procedures for inactivating human and animal prions. We provide new data concerning the efficacy of hydrogen peroxide against prions from in vitro or in vivo tests, focusing on the following: the efficiency of hydrogen peroxide sterilization and possible interactions with enzymatic or alkaline detergents, differences in the efficiency of this treatment against different prion strains, and the influence of contaminating lipids. We found that gaseous hydrogen peroxide decreased the infectivity of prions and/or the level of the protease-resistant form of the prion protein on different surface materials. However, the efficiency of this treatment depended strongly on the concentration of hydrogen peroxide and the delivery system used in medical devices, because these effects were more pronounced for the new generation of Sterrad technology. The Sterrad NX sterilizer is 100% efficient (0% transmission and no protease resistant form of the prion protein signal detected on the surface of the material for the mouse-adapted bovine spongiform encephalopathy 6PB1 strain and a variant Creutzfeldt-Jakob disease strain). Thus, gaseous or vaporized hydrogen peroxide efficiently inactivates prions on the surfaces of medical devices. PMID- 19563266 TI - Data structures for parsimony correlation and biosequence co-evolution. AB - We give an algorithm for discovering co-evolution in biosequences from a dataset consisting of aligned data and a phylogeny. The method correlates vectors of parsimony scores on the edges of a graph, averaged over all optimally parsimonious reconstructions of the data. We describe an efficient data structure, and a preprocessing step that allows for rapid, interactive computation of many correlation scores, at the expense of storage space. PMID- 19563267 TI - The search for new COX-2 inhibitors: a review of 2002 - 2008 patents. AB - BACKGROUND: Two COX isoenzymes are known, COX-1 and COX-2, for which the main inhibitors are the NSAIDs. The common anti-inflammatory drugs (such as aspirin, ibuprofen and naproxen) all act by blocking the action of both the COX-1 and COX 2 enzymes. The COX-2 inhibitors represent a new class of drugs that do not affect COX-1 but selectively block COX-2. This selective action provides the benefits of reducing inflammation without irritating the stomach and cardiovascular effects. OBJECTIVE: This review focuses on patents published in the field during 2002 - 2008, paying particular attention to promising COX-2 inhibitors. CONCLUSION: Structural analogues of the COX-2 inhibitors celecoxib and valdecoxib, and novel potential pyridazine, triazole, indole and thione derivatives emerge as promising leads for the treatment of inflammation, pain and other diseases. PMID- 19563268 TI - Macrocyclic inhibitors of HCV NS3 protease. AB - BACKGROUND: HCV NS3 is a serine protease that plays a pivotal role in catalyzing the cleavage of the single polyprotein encoded by HCV after infection of hepatocytes. Analysis of the X-ray crystal structure of the enzyme reveals a shallow catalytic site located on the surface of the protein, which has made development of inhibitors a formidable task. Attempts to discover leads by a traditional approach of screening of compound libraries have proved futile and, therefore, researchers have adopted a structure-based drug design. Analysis of the X-ray structure of NS3 protease reveals close proximity of S(1)-S(3) and S(2) S(4) pockets. Various novel approaches have been used to design preorganized, depeptidized macrocyclic inhibitors linking the P(2)-P(4) groups and P(1)-P(3) residues. OBJECTIVE: The article summarizes efforts by various groups to develop inhibitors that bind to the active site and inhibit viral replication. METHOD: Review of recent patents and scientific literature. CONCLUSION: Macrocyclization has proved to be an effective tool for depeptidization of peptidic inhibitors with improved binding and pharmacokinetic properties. PMID- 19563269 TI - Which of three structures is AZD-3199? WO-2008104790, WO-2008096112 and WO 2008096119. AB - AstraZeneca progressed the long acting beta(2) agonist into Phase II studies for the treatment of asthma and chronic obstructive pulmonary disease in August 2008. These three applications claim specific salt forms of three different long acting beta(2) agonists, all benzothiazolone derivatives. It would seem that one claims AZD-3199 whereas the other two claim backup compounds. PMID- 19563271 TI - Highlight: Perspectives in glycobiology. PMID- 19563270 TI - Drug delivery to the lymphatic system: importance in future cancer diagnosis and therapies. AB - Cancer is the second leading cause of death in the US. Currently, protocols for cancer treatment include surgery to remove diseased and suspect tissues, focused radiation, systemic chemotherapy, immunotherapy and their combinations. With conventional chemotherapy, it is almost impossible to deliver anticancer drugs specifically to the tumor cells without damaging healthy organs or tissues. Over the past several decades, efforts have been made to improve drug delivery technologies that target anticancer drugs specifically to tumor cells. It has been known for over four decades that the lymphatics are the first site of metastasis for most solid cancers; however, few efforts have been made to localize chemotherapies to lymphatic tissues. Trials of several systemic targeted drug delivery systems based on nanoparticles containing chemotherapeutic agents (e.g., liposomal doxorubicin) have shown similar antitumor activity but better patient tolerance compared with conventional formulations. Animal studies have demonstrated that nanoparticles made of natural or synthetic polymers and liposomal carriers have higher accumulation in the lymph nodes and surrounding lymphatics compared to conventional intravenous therapies. This combination has the potential to both reduce nonspecific organ toxicities and increase the chemotherapeutic dose to the most likely sites of locoregional cancer metastasis. PMID- 19563272 TI - Antiviral therapy in compensated and decompensated cirrhotic patients with chronic HCV infection. AB - Liver cirrhosis secondary to HCV infection is a chronic disorder that carries high morbidity and mortality. Approved antiviral treatment for this condition at present includes peginterferon in combination with ribavirin. Treatment is only recommended for a well-compensated liver cirrhosis, whereas antiviral therapy is commonly not implemented in cirrhotics with signs of liver decompensation, over the concern that the use of peginterferon and ribavirin might expose patients to severe treatment-related side effects. This review focuses on data available to support both efficacy and safety of antiviral therapy in both compensated and decompensated cirrhotic patients. PMID- 19563273 TI - Comprehensive management of peritoneal surface malignancy using cytoreductive surgery and perioperative intraperitoneal chemotherapy: the Washington Cancer Institute approach. AB - The peritoneal surfaces are a common site of surgical treatment failure for gastrointestinal and ovarian cancer. Progressive cancer growth at this site will result in the patient's cancer-related death. Clinical and laboratory research support new technologies to prevent disease spread to peritoneum or to treat established disease at these sites. This new treatment strategy involves a multimodality approach that combines cytoreductive surgery with perioperative intraperitoneal chemotherapy irrigations. The surgery utilizes the new surgical technique of peritonectomy, which strips away the lining of the body cavity using ball-tip electrosurgery. The surgery removes the primary cancer and any dissemination within the peritoneal cavity; the goal of the chemotherapy solution is to eradicate the microscopic residual disease. Multiple reports to date document marked survival benefits; a previously lethal condition can be cured in selected patients. Further pharmacologic research and more knowledgeable patient selection for treatment are required. PMID- 19563274 TI - Postmenopausal hormone therapy and the risk of cardiovascular disease. AB - BACKGROUND: Risks and benefits of postmenopausal hormone therapy remain highly controversial. After publication of the Women's Health Initiative hormone trials and several other major trials, the American Heart Association designated postmenopausal hormone therapy as 'Class III', if initiated for the purpose of cardiovascular disease prevention. Subsequent post hoc analyses of the Women's Health Initiative data have renewed enthusiasm for hormone therapy among younger postmenopausal women. OBJECTIVE: To review data from randomized clinical trials that have assessed cardiovascular outcomes of hormone therapy including coronary heart disease, stroke, peripheral arterial disease, and venous thromboembolism. METHODS: The review focuses on cardiovascular effects of hormone therapy only and does not attempt to integrate potential risks and benefits related to symptoms, cancer, osteoporosis or other noncardiovascular effects of postmenopausal hormone therapy. The literature search included original trial publications, post hoc analyses, and aggregate data from meta-analyses published in English and accessible to the author in full-text format for detailed analysis. RESULTS/CONCLUSION: Risks of hormone therapy seem to predominate among older postmenopausal women. Data among younger women close to menopause are insufficient to recommend such therapy for cardiovascular disease prevention. PMID- 19563275 TI - Candesartan: widening indications for this angiotensin II receptor blocker? AB - Candesartan cilexetil is one of a number of drugs of the angiotensin II receptor blocker (ARB) class. Their principal mode of action involves competitive blockade of the angiotensin II type 1 receptor, thereby modulating the activity of the rennin-angiotensin-aldosterone system. Angiotensin II receptor blocker therapy has been proven to be well tolerated and effective in the management of hypertension, chronic heart failure with left ventricular dysfunction and the prevention and progression of diabetic renal disease. Candesartan is a highly potent, long-acting and selective angiotensin II type 1 receptor blocker. It was launched in 1998 for the treatment of hypertension. Its use has increased dramatically, with recently published data suggesting benefit in the treatment of stroke, heart failure, diabetic renal disease and most recently in preventing the development of or delaying the progression of diabetic retinopathy. In this article we review the literature on the use of ARB drugs in general before focusing on candesartan. PMID- 19563276 TI - Editorial: a call to action. PMID- 19563277 TI - The American Journal of Cardiology and Journal of Periodontology editors' consensus: periodontitis and atherosclerotic cardiovascular disease. AB - ACKNOWLEDGMENT: This Editors' Consensus is supported by an educational grant from Colgate-Palmolive, Inc., New York, New York, and is based on a meeting of the authors held in Boston, Massachusetts, on January 9, 2009. DISCLOSURE: Dr. Friedewald has received honoraria for speaking from Novartis, East Hanover, New Jersey. Dr. Kornman is a full-time employee and shareholder of Interleukin Genetics, Waltham, Massachusetts, which owns patents on genetic biomarkers for chronic inflammatory diseases. Dr. Genco is a consultant to Merck, Whitehouse Station, New Jersey. Dr. Ridker has received research support from AstraZeneca, Wilmington, Delaware; Novartis; Pfizer, New York, New York; Roche, Nutley, New Jersey; Sanofi-Aventis, Bridgewater, New Jersey; and Abbott Laboratories, Abbott Park, Illinois. Dr. Ridker has received non-financial research support from Amgen, Thousand Oaks, California. Dr. Ridker is a co-inventor on patents held by Brigham and Women's Hospital that relate to the use of inflammatory biomarkers in cardiovascular disease. Dr. Ridker is a research consultant for Schering-Plough, Kenilworth, New Jersey; Sanofi-Aventis; AstraZeneca; Isis, Carlsbad, California; Novartis; and Vascular Biogenics, Tel Aviv, Israel. Dr. Van Dyke is a co-inventor on patents held by Boston University, Boston, Massachusetts, that relate to inflammation control, including consulting fees. Dr. Roberts has received honoraria for speaking from Merck, Schering-Plough, AstraZeneca, and Novartis. All other individuals in a position to control content disclosed no relevant financial relationships. PMID- 19563278 TI - Re: effect of microthreads and platform switching on crestal bone stress levels: a finite element analysis. PMID- 19563280 TI - Re: Journal of Periodontology backfiles. PMID- 19563283 TI - The effect of a pulsed Nd:YAG laser in non-surgical periodontal therapy. AB - BACKGROUND: Laser treatment may serve as an alternative or adjunctive treatment to conventional mechanical therapy in periodontics. The neodymium-doped:yttrium, aluminum, and garnet (Nd:YAG) laser has been used in periodontics. Systematic reviews aid in clinical decision-making. The aim of this study was to evaluate, in a systematic manner and after a comprehensive search of the literature, the (additional) therapeutic effects of using a pulsed Nd:YAG laser in the initial treatment of patients with periodontitis. METHODS: The focused question for this systematic review was, "What is the efficacy of a pulsed Nd:YAG laser in the initial treatment of patients with periodontitis, either as monotherapy or as an adjunct to non-surgical periodontal treatment?" The efficacy of this technique was compared to conventional therapy (ultrasonics and/or hand instrumentation) in removing plaque, as well as in improving clinical parameters, such as periodontal inflammation and probing depth. Articles published before May 2008 in the databases of MEDLINE/PubMed and the Cochrane Central Register of Controlled Trials were searched to identify appropriate studies. Clinical parameters of periodontal inflammation, namely, plaque, bleeding, gingivitis, probing depth, clinical attachment level, and gingival recession, were selected as outcome variables. RESULTS: Independent screening of the titles and abstracts of 285 PubMed and 38 Cochrane articles resulted in eight publications that met the eligibility criteria. Means +/- SDs were collected by data extraction; only descriptive analysis was possible. CONCLUSIONS: The majority of the studies analyzed showed no beneficial effect of a pulsed Nd:YAG laser compared to conventional therapy (ultrasonics and/or hand instrumentation) in the initial treatment of patients with periodontitis. The pulsed Nd:YAG laser was assessed as monotherapy and as an adjunct to non-surgical periodontal treatment; efficacy was determined by the extent of plaque removal and the reduction of periodontal inflammation. This literature review suggests that there is no evidence to support the superiority of the Nd:YAG laser over traditional modalities of periodontal therapy. PMID- 19563284 TI - A pilot study of glycosylated hemoglobin levels in periodontitis cases and healthy controls. AB - BACKGROUND: Periodontitis is associated with glycemic control in patients with diabetes. The purpose of this study was to determine if glycosylated hemoglobin is elevated in patients with periodontitis who have not been diagnosed with diabetes. METHODS: Glycosylated hemoglobin (HbA1c) was assessed using a chairside test in 59 adults without diabetes but with periodontitis (having at least five teeth with probing depth [PD] > or =5 mm, bleeding on probing [BOP], and clinical attachment or radiographic bone loss) and 53 healthy controls (PDs < or =4 mm and BOP < or =15%). Groups were compared using the t test and linear regression. Patients with HbA1c levels > or =6% were compared using the Fisher exact test and logistic regression. RESULTS: Periodontitis cases were more likely than controls to be male (68% versus 38%; P = 0.002) and current or former smokers (P = 0.002). Cases had significantly higher body mass index (BMI) than controls (27.6 kg/m(2) versus 25.5 kg/m(2); P = 0.018) but were of similar age (51.3 years versus 50.9 years; P = 0.89). Unadjusted mean HbA1c levels did not differ significantly between cases and controls (5.66% +/- 0.56% versus 5.51% +/- 0.44%; P = 0.12). After adjustments for age, gender, BMI, and current smoking, mean HbA1c was significantly higher in cases (between-group difference, 0.21%; P = 0.046). A higher proportion of cases (27.3%) than controls (13.2%) had HbA1c values > or =6%, although this difference was not statistically significant (P >0.1). CONCLUSIONS: Periodontitis is associated with a slight elevation in glycosylated hemoglobin. The clinical significance of this difference remains to be determined. This preliminary finding is consistent with earlier reports that periodontitis is associated with elevated blood glucose in adults without diabetes and may increase one's risk for type 2 diabetes. PMID- 19563286 TI - Prevalence of implant loss and the influence of associated factors. AB - BACKGROUND: The objective of this study was to assess the outcome of dental implants inserted at the Institute of Clinical Odontology, University of Oslo, between 1990 and 2005. The prevalence of implant loss and the factors associated with the outcome were studied. METHODS: A total of 164 subjects were invited to participate in this cross-sectional project, of whom 55 were unable, leaving 109 volunteers available for examination. The study population included 69 females and 40 males with a mean age of 43.8 years at the time of implant insertion (range, 18 to 80 years). At the subject level, the mean time from implant loading to the present examination was 8.4 years (range, 1.1 to 16.0 years). The participants were examined clinically and radiographically and interviewed regarding general health and habits. RESULTS: The 109 examined subjects had been treated with 374 implants. Eighteen implants (4.8%) were lost in 10 subjects (9.2%). Eleven implants were lost before loading, three were lost during the first 5 years after loading, and four were lost 5 to 10 years after loading. No implants were lost after >10 years of loading. The loss of oral implants was significantly associated with a history of smoking and periodontitis (P <0.05). CONCLUSIONS: The inserted implants showed a high survival rate, especially after the first year of insertion, even though the subjects were not maintained by specialists. All late implant losses were preceded by an early loss. Implant loss was significantly associated with smoking and periodontitis. PMID- 19563285 TI - Periodontal disease status in gullah african americans with type 2 diabetes living in South Carolina. AB - BACKGROUND: African Americans have a disproportionate burden of diabetes. Gullah African Americans are the most genetically homogeneous population of African descent in the United States, with an estimated European admixture of only 3.5%. This study assessed the previously unknown prevalence of periodontal disease among a sample of Gullah African Americans with diabetes and investigated the association between diabetes control and the presence of periodontal disease. METHODS: Two hundred thirty-five Gullah African Americans with type 2 diabetes were included. Diabetes control was assessed by percentage of glycosylated hemoglobin (HbA1c) and divided into three categories: well controlled, <7%; moderately controlled, 7% to 8.5%; and poorly controlled, >8.5%. Participants were categorized as healthy (no clinical attachment loss [AL] or bleeding on probing) or as having early periodontitis (clinical AL > or =1 mm in at least two teeth), moderate periodontitis (three sites with clinical AL > or =4 mm and at least two sites with probing depth [PD] > or =3 mm), or severe periodontitis (clinical AL > or =6 mm in at least two teeth and PD > or =5 mm in at least one site). Observed prevalences of periodontitis were compared to rates reported for the National Health and Nutrition Examination Survey (NHANES) studies. RESULTS: All subjects had evidence of periodontal disease: 70.6% had moderate periodontitis and 28.5% had severe disease. Diabetes control was not associated with periodontal disease. The periodontal disease proportions were significantly higher than the reported national prevalence of 10.6% among African Americans without diabetes. CONCLUSION: Our sample of Gullah African Americans with type 2 diabetes exhibited a higher prevalence of periodontal disease compared to African Americans, with and without diabetes, as reported in NHANES III and NHANES 1999 2000. PMID- 19563287 TI - Semilunar coronally positioned flap or subepithelial connective tissue graft for the treatment of gingival recession: a 30-month follow-up study. AB - BACKGROUND: The objective of this prospective, controlled clinical trial was to evaluate the long-term outcomes of subepithelial connective tissue graft (SCTG) or semilunar coronally positioned flap (SCPF) for the treatment of Miller Class I gingival recession defects. METHODS: Seventeen patients with bilateral Miller Class I gingival recessions (< or =4.0 mm) in maxillary canines or premolars were selected. The recessions were randomly assigned to receive SCPF or SCTG. Recession height (RH), recession width (RW), width of keratinized tissue (WKT), thickness of keratinized tissue (TKT), probing depth (PD), and clinical attachment level (CAL) were measured at baseline and at 6 and 30 months post surgery. Patient satisfaction with esthetics and root sensitivity was also evaluated. RESULTS: The root-coverage outcomes obtained at 6 months were maintained throughout the study. At the 30-month examination, the average percentage of root coverage was 89.25% for SCPF and 96.83% for SCTG (P >0.05); complete root coverage was observed in 58.82% and 88.24% of patients, respectively. SCTG maintained a statistically significant increase in TKT (P <0.05) at 30 months. At this time, there were no significant differences between the two groups with regard to RH, RW, WKT, PD, and CAL. The evaluation of the esthetic outcome by the patient showed a preference for the SCTG treatment. Furthermore, in this group, no patient complained of residual or additional root hypersensitivity. In the SCPF group, three patients had this complaint at 30 months. CONCLUSIONS: SCPF and SCTG can be successfully used to treat Class I gingival recession, presenting outcomes with long-term stability. However, patient-oriented outcomes, such as esthetics and root sensitivity, favor SCTG therapy. PMID- 19563288 TI - Coronally advanced flap with and without vertical releasing incisions for the treatment of multiple gingival recessions: a comparative controlled randomized clinical trial. AB - BACKGROUND: Vertical releasing incisions might damage the blood supply to the flap and result in unesthetic visible white scars. The aim of the present study was to compare root coverage and esthetic outcomes of the coronally advanced flap (CAF) with and without vertical releasing incisions in the treatment of multiple gingival recessions. METHODS: Thirty-two systemically and periodontally healthy subjects with esthetic complaints due to the exposure of Miller Class I and II multiple (at least two) gingival recession defects (> or =1 mm) affecting adjacent teeth of the same quadrant of the upper jaw were enrolled in the study. Sixteen patients (with 45 gingival recessions) were randomly assigned to the control group, and the other 16 patients (with 47 recession defects) were assigned to the test group. All recessions were treated with a CAF; vertical releasing incisions were performed in the control group, whereas an envelope-type flap was used in the test group. The patient's postoperative morbidity was assessed 1 week after the surgery, whereas the esthetic evaluation, made by the patient and independent periodontist, and the clinical evaluation were made 1 year later. RESULTS: Surgical time was significantly shorter in the envelope-type CAF group. No statistically significant difference was demonstrated between the two groups in terms of recession reduction and clinical attachment level gain. A statistically greater probability of complete root coverage (adjusted odds ratio, 3.76; 95% confidence interval: 0.92 to 15.33; P <0.05) and a greater increase in buccal keratinized tissue height were observed with the envelope type of CAF. Patient satisfaction with esthetics was very high in both treatment groups, with no statistically significant differences between them. A better postoperative course and better results in the esthetic evaluation made by an independent periodontist were demonstrated in patients treated with the envelope type of CAF. CONCLUSIONS: Both CAF techniques were effective in reducing recession depth. The envelope type of CAF was associated with an increased probability of achieving complete root coverage and with a better postoperative course. Keloid formation along the vertical releasing incisions was responsible for the worst esthetic evaluation made by an independent expert periodontist. PMID- 19563289 TI - The role of the composite interleukin-1 genotype in the association between periodontitis and acute myocardial infarction. AB - BACKGROUND: Recent data indicate that interleukin (IL)-1 polymorphism may influence the susceptibility to periodontitis and coronary heart diseases. The aim of this study was to evaluate the impact of the composite IL-1 genotype (allele 2 at IL-1A -889 and IL-1B +3954) in the association between acute myocardial infarction (AMI) and periodontitis. METHODS: One hundred four white subjects (54 patients with AMI and 50 healthy controls) were studied; each received a comprehensive periodontal examination, including measurement of periodontal probing depth (PD) and clinical attachment level (CAL). The extent of periodontitis was assessed by the percentage of sites with clinical AL >3 mm. Polymorphisms in the IL-1 gene cluster were assessed using a reverse hybridization assay. RESULTS: Compared to controls, mean values for PD (4.6 mm versus 3.7 mm; P <0.0001) and CAL (5.4 mm versus 4.5 mm; P = 0.0001) were significantly increased among patients with AMI. Significantly more subjects with moderate or severe periodontitis (> or =33% of sites with clinical AL >3 mm) were found in the AMI group compared to controls (31.5% versus 8%; P = 0.0016). These differences remained statistically significant after adjustment for smoking, age, and gender. No significant differences were observed in the allele frequencies of the gene loci IL-1A -889 and IL-1B +C3954 between patients with AMI and controls. Also, there was no difference in the frequency of the composite IL-1 genotype. IL 1 genotype-positive patients with AMI had slightly increased PD and AL compared to IL-1 genotype-negative patients with AMI. CONCLUSIONS: The results confirmed an association between periodontitis and AMI but failed to detect a modifying impact of the composite IL-1 genotype. Although the IL-1 genotype was only weakly associated with compromised periodontal health, it was not associated with AMI. PMID- 19563290 TI - 1-Tetradecanol complex: therapeutic actions in experimental periodontitis. AB - BACKGROUND: The present study was planned to investigate the therapeutic actions of 1-tetradecanol complex (1-TDC), a novel monounsaturated fatty acid mixture, in established periodontitis in rabbits. METHODS: Periodontitis was initiated in 18 New Zealand White rabbits using ligatures around mandibular second premolars, followed by topical Porphyromonas gingivalis application (10(9) colony forming units). After 6 weeks of disease induction (phase 1), three animals were sacrificed to assess the established periodontitis level. P. gingivalis application was discontinued, and the remaining 15 animals continued with topical treatment of 1-TDC (100 mg/ml; n = 5) or placebo (n = 5) or no treatment (n = 5) for an additional 6 weeks (phase 2). Mandibular block sections obtained after euthanasia were decalcified and embedded in paraffin. In addition to the macroscopic analyses, hematoxylin and eosin-stained sections were used to study cellular inflammatory infiltrate and quantitative histomorphometry. Tartrate resistant acid phosphatase and osteocalcin were used to identify osteoclastic and osteoblastic activity, respectively. RESULTS: P. gingivalis application resulted in periodontal disease with gingival inflammation and bone loss (30% compared to baseline) at 6 weeks. Treatment with 1-TDC stopped the progression of the disease and resulted in a significant reduction in the macroscopic periodontal inflammation, attachment, and bone loss (10.1% +/- 1.8%), whereas periodontal disease progressed in the untreated and placebo groups (P <0.05). Histologic assessment and histomorphometric measurements demonstrated that 1-TDC inhibited inflammatory cell infiltration and osteoclastic activity (P <0.05). CONCLUSION: The findings suggest that topical application of cetylated monounsaturated fatty acid complex (1-TDC) is a potential therapeutic approach in controlling the progression of chronic periodontal disease. PMID- 19563291 TI - Elevated plasma calcifediol is associated with aggressive periodontitis. AB - BACKGROUND: Vitamin D is associated with a number of inflammatory diseases and plays a significant role in regulating bone metabolism. Serum calcifediol was demonstrated to be potentially associated with periodontal disease. The purpose of this study was to evaluate whether an association exists between plasma calcifediol concentrations and aggressive periodontitis (AgP) and whether plasma levels of bone-related biomarkers (osteocalcin, alkaline phosphatase, calcium, and phosphorus) regulated by vitamin D are related to AgP. METHODS: Sixty-six patients with generalized AgP, 52 patients with chronic periodontitis, and 60 healthy controls were included in this study. Periodontal examination consisted of probing depth, attachment loss, and bleeding index measurements. Hematic calcifediol and bone-related biomarker levels were detected using radioimmunity assay kits or a biochemical analyzer. RESULTS: Plasma calcifediol levels in patients with AgP were higher than those of healthy controls (29.28 versus 21.60 nmol/l; P <0.05) and were statistically significantly correlated with bleeding index (r = 0.321; P <0.05). Plasma osteocalcin concentrations in patients with AgP were higher than those of healthy controls (0.90 versus 0.70 ng/ml; P <0.05). Serum inorganic phosphorus values of both periodontitis groups were lower than those of healthy controls (1.06 +/- 0.18 mmol/l and 1.10 +/- 0.15 mmol/l versus 1.26 +/- 0.17 mmol/l; P <0.05). CONCLUSION: Plasma calcifediol levels might be associated with periodontal inflammation. PMID- 19563292 TI - The influence of atopy in the prognosis of the replantation of avulsed teeth. AB - BACKGROUND: Replantation is the most indicated procedure for traumatic dental avulsion, but it invariably results in a greater or lower degree of root resorption. The outcomes of these cases can be affected by some well-known factors. Because studies have shown the importance of the innate and acquired immune response in the mechanisms involved in the resorption of bone tissues, an endeavor was made to verify if such an influence applies to dental resorption. METHODS: Fifty-seven avulsed, endodontically treated teeth were assessed in the Endodontics Clinic, Pontifical Catholic University of Parana. Follow-up of the replanted teeth included periodical clinical examinations and radiographs, in accordance with the control rules of the International Association of Dental Traumatology. The evaluation of atopy was based on the patient's personal and family history in conjunction with the skin-prick test for five allergen extracts. RESULTS: Of the 46 teeth with a favorable outcome, 33 (71.74%) were in atopic patients, and 13 (28.26%) were in non-atopic patients. Of the 11 teeth with an unfavorable outcome, four (36.36%) were in atopic patients, and seven (63.64%) were in non-atopic patients; a greater prevalence of unfavorable prognoses occurred in non-atopic patients. CONCLUSION: The outcome after 1 year for avulsed and replanted teeth is more favorable in atopic patients. PMID- 19563293 TI - Bone strain and interfacial sliding analyses of platform switching and implant diameter on an immediately loaded implant: experimental and three-dimensional finite element analyses. AB - BACKGROUND: Strain gauge analysis and finite element (FE) simulations were used to estimate the bone strain and micromovement at the bone-implant interface (BII) for platform switching and different diameters of a single, immediately loaded mandibular implant. METHODS: Four models were created, including 5-mm-diameter implants assembled with abutments that were 5 or 4 mm in diameter on bonded (delay-loading treatment) and contact (immediate-loading treatment) BIIs; a model with an implant diameter of 3.75 mm was also analyzed. Vertical and lateral loads of 130 N were applied to all models. RESULTS: During lateral loading, the strains were highly concentrated on one side of the mandible in the experimental and validation FE models. Bone strains were reduced by <10% when platform switching was used compared to no platform switching. However, increasing implant diameter decreased the surrounding bone strain significantly. The sliding and gap distances at the BII did not differ significantly among all models considered. CONCLUSIONS: Bone strain was reduced more by increasing the diameter of the implant than by using platform switching in the immediately loaded implant. However, neither a wide implant nor platform switching reduced micromotion at the BII for enhancing implant stability. PMID- 19563294 TI - Real-time polymerase chain reaction to determine the prevalence and copy number of epstein-barr virus and cytomegalovirus DNA in subgingival plaque at individual healthy and periodontal disease sites. AB - BACKGROUND: Detection of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) in plaque from patients with periodontal disease provides support for the theory that these viruses play a role in the pathogenesis of periodontitis. This study sought to further define this relationship by determining the prevalence of these viruses at individual disease and healthy sites of patients with periodontal disease and to determine whether the presence and amount of viral DNA correlate with disease severity. METHODS: Subgingival plaque from three healthy and three disease sites of 65 patients who had chronic periodontitis were evaluated for the presence and amount of EBV, CMV, and Fusobacterium nucleatum DNA using real-time polymerase chain reaction. Patient serum was evaluated for antibodies against EBV and CMV using enzyme-linked immunosorbent assays. RESULTS: EBV DNA was detected in 18.5% of subgingival plaque samples (72/390) and in at least one of the six plaque samples in 44.6% (29/65) of the patients. CMV DNA was detected in one plaque sample (0.3%). EBV was significantly more prevalent in disease sites (28.2%; 55/195) than in healthy sites (8.7%; 17/195; P = 0.002). However, neither EBV prevalence nor its amount correlated with increased probing depth >5 mm or attachment loss >2 mm, whereas the amount of F. nucleatum DNA did. Sites positive for EBV had a median copy number of eight. Antibodies against EBV and CMV were detected in 85.7% and 78.6% of persons evaluated, respectively. CONCLUSION: EBV was infrequent and CMV was rarely present in individual subgingival sites affected by chronic periodontitis. PMID- 19563295 TI - B-1a cells in experimental gingivitis in humans. AB - BACKGROUND: Although previous studies revealed the presence of autoreactive B cells (B-1a cells) in periodontitis lesions, no evidence was provided for an active role of such cells in the host response to microbial challenge. The aim of the present investigation was to study the reaction of B-1a cells to de novo plaque formation in subjects who were treated for severe chronic periodontitis. METHODS: Fifteen white subjects with generalized, severe chronic periodontitis volunteered. Surgical periodontal therapy was performed in all quadrants of each subject after a period of infection control. After 6 months of healing (baseline), two gingival biopsies were harvested from each patient (probing depth <4 mm and no bleeding on probing; healed sites). The experimental gingivitis model was applied, and plaque accumulation was allowed for 3 weeks. Two additional biopsies were collected and prepared for immunohistochemical analysis on day 21. RESULTS: The biopsies retrieved after 3 weeks of plaque accumulation contained larger proportions of CD19+ and CD5+ cells (B-1a cells) than biopsies representing baseline (healed sites) (7.38% +/- 2.80% versus 5.96% +/- 2.48%). The tissue fraction of cells carrying the markers for CD3 (T cells), CD19 (B cells), and Bcl2 (apoptosis-associated marker) were significantly larger in tissue samples collected after 3 weeks of plaque accumulation than in specimens from baseline (healed sites). CONCLUSION: Autoreactive B cells (B-1a cells) are involved in the host response to microbial challenge in subjects with chronic periodontitis. PMID- 19563296 TI - Within-subject variability in repeated measures of salivary analytes in healthy adults. AB - BACKGROUND: Saliva contains a large number of biomolecules, some of which have putative diagnostic usefulness. A potential problem with the use of biomolecules in diagnosis is day-to-day fluctuation due to within-subject variability. This study evaluated the intraindividual variability of six salivary analytes in healthy adults and determined their normal range. METHODS: Unstimulated whole saliva (5 ml) was collected every 2 to 3 days on six occasions from 30 subjects in good oral and systemic health. Four of the samples were collected in the clinic, and two were collected by the subject at home. The concentration ranges of interleukin (IL)-1beta, IL-6, matrix metalloproteinase-8, prostaglandin E(2), tumor necrosis factor-alpha, interferon-alpha, and albumin were examined. Descriptive statistics were computed, and a one-way random-effects model was used to quantify within- and between-subject components of variability. Intraclass correlation coefficients (ICCs) were calculated for each subject/analyte combination. RESULTS: Within-subject coefficients of variation for these analytes ranged from 67.6% to 172.1% for the in-clinic samples and from 111.9% to 201.0% for the at-home samples. The ICC for the various analytes ranged from 41% to 61% for the in-clinic samples. The at-home samples exhibited significantly more variability than did those obtained in the clinic under supervision. CONCLUSIONS: There was marked within-subject variation in the salivary concentrations of these analytes. With increased interest in salivary diagnostics, the within-subject variability, normal range, and threshold levels for abnormal levels of individual salivary analytes need to be determined if these diagnostics tests are to have clinical usefulness. PMID- 19563297 TI - Periodontitis in pregnancy: clinical and serum antibody observations from a baboon model of ligature-induced disease. AB - BACKGROUND: Chronic oral infections that elicit host responses leading to periodontal disease are linked with various sequelae of systemic diseases. This report provides seminal information on the clinical and adaptive immunologic characteristics of a baboon model of ligature-induced periodontitis during pregnancy. METHODS: Female Papio anubis were evaluated for periodontal health at baseline. Ligatures were tied around selected teeth to initiate oral inflammation and periodontitis. Then the animals were bred. At midpregnancy ( approximately 90 days), a clinical evaluation was performed, and additional ligatures were tied on teeth in the contralateral quadrants to maintain progressing periodontitis throughout pregnancy. A final clinical evaluation was done for all experimental teeth after delivery, and ligatures were removed. Serum was collected at all sampling intervals for the determination of antibody levels to a group of 20 oral bacteria. Unligated animals served as controls. RESULTS: At baseline, 16% of animals exhibited minimal plaque and gingival inflammation without periodontal disease. The remaining baboons demonstrated varying levels of inflammation/bleeding, and approximately 20% of the population had periodontal pocketing (>3 mm). Ligated animals expressed increased levels of inflammation and increased probing depths and clinical attachment loss (AL) and could be stratified into multiple subsets postligation based upon changes in clinical parameters at midpregnancy and at delivery. Baboons were categorized into disease susceptibility groups (periodontal disease susceptibility 1 through 4) that described the extent/severity of induced disease during pregnancy. Control animals showed minimal periodontal changes during gestation. Significant differences in serum antibody to multiple oral bacteria were found in animals presenting with periodontitis at baseline and during the 6 months of ligature induced disease. A significant correlation to antibody to P. gingivalis, which was sustained throughout ligation and pregnancy, was observed with disease presentation. CONCLUSIONS: The clinical presentation at baseline, reflecting the natural history of oral disease in these animals, suggests individual variation that is reflected in the characteristics of the adaptive immune responses to oral bacteria. The variability in the response to ligation with resulting periodontal disease provides a model to document prospectively the relationship between oral and systemic health outcomes. PMID- 19563298 TI - Advanced glycation end products inhibit the expression of collagens type I and III by human gingival fibroblasts. AB - BACKGROUND: It is evident that diabetes and periodontal disease are closely interrelated. Accumulation of advanced glycation end products (AGEs), coupled with exaggerated host responses to bacterial infection, may account for the increased periodontal destruction observed in patients with uncontrolled diabetes. The present study investigated the effects of AGEs on the viability of human gingival fibroblasts (HGFs) and the expression of types I and III collagen in HGFs. METHODS: The cell viability of HGFs was examined by methylthiazolet etrazolium assay, whereas the expression of types I and III collagen message and protein was detected by real-time quantitative reverse transcription-polymerase chain reaction and sandwich enzyme-linked immunosorbent assay, respectively. RESULTS: AGEs significantly suppressed the cell viability of HGFs from 24 to 72 hours (P <0.01). A high concentration of glucose (25 mmol/l) in the culture media exaggerated the inhibition of the survival rate of HGFs (P <0.01). The expression of collagen types I and III messages and proteins was significantly downregulated at 72 hours by AGEs in a concentration-dependent manner (P <0.05). Moreover, the synthesis of intracellular types I and III collagen protein was markedly inhibited by AGEs (P <0.05). CONCLUSIONS: AGEs may suppress the cell viability of HGFs and downregulate the expression of types I and III collagen by the cells. Further investigations are warranted to clarify the molecular mechanisms of AGEs in the regulation of cell function and collagen metabolism in patients with diabetes and periodontitis. PMID- 19563299 TI - Fusobacterium nucleatum enters normal human oral fibroblasts in vitro. AB - BACKGROUND: Fusobacterium nucleatum, a commensal opportunistic oral bacterium, is capable of invading gingival epithelial cells, but the entrance into human primary oral fibroblast cells has not been documented. This study evaluated the ability of three strains of F. nucleatum (F. nucleatum ssp. nucleatum, F. nucleatum ssp. polymorphum, and F. nucleatum ssp. vincentii) to enter gingival fibroblasts (GFs) and periodontal ligament fibroblasts (PLFs). METHODS: GFs and PLFs were cocultured for various periods of time with different strains of F. nucleatum. Scanning and transmission electron microscopy, together with confocal laser scanning microscopy, were used to visualize the entrance and presence of bacteria in host cells. Flow cytometry was performed to compare the load of internalized bacteria in GFs and PLFs exposed for 3 and 5 hours to live F. nucleatum labeled with fluorescein isothiocyanate. RESULTS: All three strains of F. nucleatum were found entering and located in the cytoplasm of GFs and PLFs after 1 hour of exposure. Flow cytometry tests revealed a significant increase in the fluorescent signal, compared to baseline, derived from bacteria internalized in fibroblasts exposed for 3 hours (P <0.001); a further increase was found at 5 hours. The greatest bacterial mass in exposed fibroblasts of both types was of F. nucleatum ssp. polymorphum; the smallest was of F. nucleatum ssp. vincentii. Although not statistically significant, PLFs had a higher bacterial load than corresponding GFs. CONCLUSION: F. nucleatum was capable of entering GFs and PLFs in a manner that is dependent on the cell type and the bacterial strain. PMID- 19563301 TI - Guided tissue regeneration-based root coverage with a platelet concentrate graft: a 3-year follow-up case series. AB - BACKGROUND: A platelet concentrate graft (PCG) was applied underneath the barrier membrane in guided tissue regeneration (GTR)-based root coverage procedures. The purpose of this case study was to assess the effectiveness of the technique and the long-term outcomes. METHODS: Thirty-seven Miller Class I or II recession defects in six patients were treated using the PCG and a collagen membrane covered by an advanced mucogingival flap. Clinical parameters, such as vertical gingival recession depth (VRD), clinical attachment level (CAL), probing depth (PD), and width of keratinized tissue (WKT), were recorded presurgery (at baseline [BL]) and at 6 and 36 months of follow-up. The Wilcoxon signed-rank test and the Mann-Whitney test were used to assess statistical significance (alpha = 0.05). RESULTS: Between BL and 6 months, there was a statistically significant reduction in VRD (from 2.81 +/- 0.88 mm to 0.30 +/- 0.48 mm) and PD (from 2.59 +/ 0.50 mm to 1.14 +/- 0.35 mm), as well as an improvement in CAL (from 5.41 +/- 0.86 mm to 1.43 +/- 0.66 mm). These results remained stable over the following 30 months. There was no statistically significant change in WKT during the initial 6 months; however, the increase in WKT was statistically significant at the 36 month follow-up. When comparing the healing between the maxillary and mandibular teeth at 6 months, the VRD reduction was statistically significantly greater in the maxillary teeth, but the difference was not statistically significant 30 months later. The average root coverage at 6 and 36 months was 89.86% +/- 15.85% and 85.86% +/- 18.16%, respectively, with complete root coverage in 24 (64.86%) and 21 (56.76%) of the 37 teeth. CONCLUSIONS: The GTR-based technique using PCG was effective in reducing gingival recession. The outcomes remained stable for 3 years. PMID- 19563300 TI - Clinical outcomes of peri-implant peripheral giant cell granuloma: a report of three cases. AB - BACKGROUND: Peripheral giant cell granuloma (PGCG) is a reactive lesion that occurs on the gingiva or alveolar mucosa and contains numerous giant cells. Its recurrence rate is 10%. Only five cases associated with dental implants have been reported. This case report describes three additional cases with clinical courses and outcomes. METHODS: Three women presented with a chief complaint of a gingival mass around the implants. The lesions were surgically excised under local anesthesia. RESULTS: The initial diagnosis at presentation was pyogenic granuloma. Radiography showed marginal bone loss accompanying the lesions. Histopathology confirmed the diagnosis of PGCG. In two cases, several recurrences resulted in explantation of the fixture. One case healed uneventfully. CONCLUSIONS: Despite its usually benign clinical behavior, peri-implant PGCG may follow an aggressive course. Treatment planning for this condition should take into account the presence of recurrences to evaluate the necessity of an aggressive surgical approach that may involve advanced bone loss and explantation. Further research on the origin of this implant-associated condition with a larger series of cases is necessary to provide a basis for adequate management. PMID- 19563302 TI - Bacteriophage-encoded functions engaged in initiation of homologous recombination events. AB - Recombination plays a significant role in bacteriophage biology. Functions promoting recombination are involved in key stages of phage multiplication and drive phage evolution. Their biological role is reflected by the great variety of phages existing in the environment. This work presents the role of recombination in the phage life cycle and highlights the discrete character of phage-encoded recombination functions (anti-RecBCD activities, 5' --> 3' DNA exonucleases, single-stranded DNA binding proteins, single-stranded DNA annealing proteins, and recombinases). The focus of this review is on phage proteins that initiate genetic exchange. Importance of recombination is reviewed based on the accepted coli-phages T4 and lambda models, the recombination system of phage P22, and the recently characterized recombination functions of Bacillus subtilis phage SPP1 and mycobacteriophage Che9c. Key steps of the molecular mechanisms involving phage recombination functions and their application in molecular engineering are discussed. PMID- 19563303 TI - Oral delivery of gastro-resistant microencapsulated typhoid vaccine. AB - Oral vaccination has long been regarded as the best alternative to conventional parenteral vaccination considering practical, economical, and immunological aspects. The purpose of this study was to develop albumin-chitosan mixed matrix microsphere-filled coated capsule formulations of Typhoid Vi antigen and to determine whether it can induce antigen-specific mucosal and systemic immune responses on oral administration. Formulations with Typhoid Vi antigen were prepared and filled into hard gelatin capsules (size # 9) and enteric coated. Formulations were characterized and administered to Sprague-Dawley rats to evaluate the induction of immune response to the antigen. The results indicated that the particle size, zeta potential, swelling, and disintegration rates were optimal for the oral delivery of microencapsulated vaccines. In vivo studies displayed multifold increase of antigen-specific IgG and IgA levels 8 weeks after oral immunization. No statistically significant difference in the antigen specific IgG and IgA levels were found between oral and parenteral injection groups 8 weeks after vaccination. On the basis of the results of the study, it can be concluded that the oral administration of Typhoid Vi antigen microspheres was successful in inducing antigen-specific systemic and mucosal immune response. PMID- 19563304 TI - Growth-differentiation factor-15, endoglin and N-terminal pro-brain natriuretic peptide induction in athletes participating in an ultramarathon foot race. AB - We investigated the actions of growth-differentiation factor (GDF)-15, endoglin and N-terminal pro-brain natriuretic peptide (NT-pro-BNP) in 15 male athletes who participated in the ultradistance foot race of the 246 km 'Sparthathlon'. Measurements were performed before (phase I), at the end of the race (phase II) and 48 h post-race (phase III). GDF-15 and endoglin serum concentrations were determined with enzyme-linked immunosorbent assay and NT-pro-BNP plasma levels by electrochemiluminescence. GDF-15 levels were increased from phase I (563.9 +/- 57.1 pg ml(-1)) to phase II (2311.1 +/- 462.3 pg ml(-1)) and decreased at phase III (862.0 +/- 158.0 pg ml(-1)) (p < 0.0002). NT-pro-BNP levels followed a similar pattern to that of GDF-15 from 38.1 +/- 4.8 pg ml(-1) at phase I to 1280.6 +/- 259.0 pg ml(-1) at phase II and 89.8 +/- 13.6 pg ml(-1) at phase III (p < 0.0001) and at the same time points, endoglin levels were 4.7 +/- 0.2 ng ml( 1) at phase I, 5.8 +/- 0.2 ng ml(-1) at phase II and 4.3 +/- 0.2 ng ml(-1) at phase III (p < 0.002). These findings indicate that circulating GDF-15, endoglin and NT-pro-BNP levels reflect a transient endothelial dysfunction in these athletes who participated in a foot race consisting of continuous, prolonged and brisk exercise. PMID- 19563305 TI - The economic crisis, the Pharmaceutical Benefits Scheme, and the dilemmas of medicines policy. PMID- 19563306 TI - PBS reform - a missed opportunity? AB - The Pharmaceutical Benefits Scheme (PBS) reform package was announced in 2006 and was designed to save the government significant expenditure on the PBS through mandatory price cuts and price disclosure arrangements for multi-brand products. Perhaps most significantly, the formulary was spilt in two with no linkages between the formularies on either price or therapeutic outcome. This article examines the potential impact of these changes on the PBS and pharmaceutical policy in Australia more broadly. PMID- 19563307 TI - The PBS in a globalised world: free trade and reference pricing. AB - In January 2005 Australia implemented the Australia-United States Free Trade Agreement (AUSFTA). The agreement had placed domestic health policy and the Pharmaceutical Benefits Scheme (PBS) in particular, on the trade negotiating table. At the time Australians were told the PBS would not be undermined, but why was it included in a trade agreement? This article argues that recent reforms to the PBS partially delivered on an issue that the US has compelled its trade negotiators to ensure since 2002: the elimination of reference pricing. In Australia, reference pricing, as used by the PBS, had been credited with obtaining money when buying new medicines. PMID- 19563308 TI - Transparency in pricing arrangements for medicines listed on the Australian Pharmaceutical Benefits Scheme. AB - Australia's system for assessing the cost-effectiveness of drugs for listing under the Pharmaceutical Benefits Scheme (PBS) is recognised internationally. A variety of mechanisms, such as evidence-based rules for determining eligibility for initial or continuing subsidy, price-volume agreements, rebates, and caps on government expenditure are used to contain PBS expenditures. In this paper we assess the extent of use of special pricing arrangements in Australia and how and where they are communicated to health professionals and the community. We searched publicly available documents published by the Pharmaceutical Benefits Advisory Committee (PBAC) and the Pharmaceutical Benefits Pricing Authority (PBPA). We found 73 medicines where special pricing arrangements had been applied and where prices appearing on the Schedule of Pharmaceutical Benefits might differ from those considered to be "cost-effective" by the PBAC. Reporting of these special pricing agreements was inconsistent and generally non-transparent. In some, the lack of transparency may have reflected the desire of manufacturers to disguise the true negotiated price, lest it weaken their negotiation position in other jurisdictions. PMID- 19563309 TI - High prices for generics in Australia - more competition might help. AB - It is commonly believed that dispensed prices of medicines in Australia are substantially lower than those in other developed countries, particularly the US. This article reports the results of an analysis comparing dispensed prices for the most commonly prescribed and the highest cost items in Australia with dispensed prices in the US. Although a large majority of items are less expensive in Australia than in the US, Australian prices are higher for a substantial number of products, particularly generic drugs. This article examines various policies affecting the pricing of generics in Australia. It is postulated that the main cause for higher prices for a substantial number of generic products is the lack of price competition. This results from government policy which ensures that a price reduction by one company is communicated immediately to all competitors in that market along with an invitation to match the reduced price. The dominant strategy for all suppliers is to only reduce their price in response to a reduction in price by a competitor. The result is a lack of differentiation in pricing across brands of a medicine on the Schedule of Pharmaceutical Benefits. The government could improve the structure of the generics market and encourage greater competition by ceasing to disclose competitor firms' offers to other competitors. The government could conduct pricing reviews of each generic product relatively infrequently (eg, only once annually or every 18 months). At the time of the pricing review, the government would request confidential offers on price for a generic from all players in the market. Brands should then all be listed under the Pharmaceutical Benefits Scheme (PBS) at the offered price. Prices offered by the individual supplier would apply until the next pricing review. The PBS would continue to subsidise up to the price of the lowest priced brand, with brand premiums applying to all brands priced higher than the benchmark price. Such an approach would provide opportunity for players in the market to capture market share by being the lowest priced brand. PMID- 19563310 TI - The impact of copayments and safety nets on PBS expenditure. AB - This article discusses the impact on patients of changes in copayments and safety net thresholds (SNT) within the Pharmaceutical Benefits Scheme (PBS). Trends in the nominal and real values of copayments and SNTs are examined, as are changes in the numbers of types of patients. The relationship between the number of safety net cardholders and copayments and SNTs is estimated. Increases in the number of copayments necessary to reach the safety net threshold restrict the number of patients able to benefit from this provision. Policy for determining the levels of copayments and safety net thresholds needs to be put on a rationale basis in a way similar to the determination of prices for PBS medicines. PMID- 19563311 TI - Australia's pharmaceutical cost sharing policy: reducing waste or affordability? AB - In this paper we argue that Australia's pharmaceutical cost sharing policy has been applied as if cost sharing is unproblematic for medicine affordability and good health outcomes. Australian and international experience with pharmaceutical cost sharing strongly suggests a negative impact on affordability and quality use of medicines, disproportionately affecting low income patients. We argue that Australia's use of cost sharing reflects the currency of a cognitively powerful and morally charged idea - moral hazard. Moral hazard refers to the change in behaviour induced by insurance coverage. Applied to pharmaceuticals, this means that low out-of-pocket cost will lead to waste. Moral hazard mixes the explanatory power of price with the intuitively cogent notion that if people do not experience consequences they will behave irresponsibly. Cost sharing policy has gone unscrutinised and uncontested not because cost sharing is unproblematic, but because in the light of the idea of moral hazard it has all the question deadening weight of common sense. PMID- 19563312 TI - PBS copayments and safety nets. PMID- 19563313 TI - Biopharmaceutical innovation and industrial developments in South Korea, Singapore and Taiwan. AB - South Korea, Singapore and Taiwan are well known as export-oriented developmental states which for decades employed industrial policy to target particular industries for government support. In the past fifteen years, these three countries all identified the biopharmaceutical industry as a strategic sector. This article explores, through economic analysis, the rationale for this decision and the strategies chosen for linking into the global bio-economy with the objective of catching up in biopharmaceuticals. The paper identifies three comparative advantages enjoyed by these countries in the biopharma sector: (1) public investments in basic research; (2) private investments in phase 1 clinical trials; and (3) a potentially significant contract research industry managing latter-stage clinical trials. Governments employ a range of industrial policies, consistent with these comparative advantages, to promote the biopharmaceutical industry, including public investment in biomedical hubs, research funding and research and development (R&D) tax credits. We argue that the most important feature of the biopharmaceutical industry in these countries is the dominant role of the public sector. That these countries have made progress in innovative capabilities is illustrated by input measures such as R&D expenditure as share of gross domestic product, number of patents granted and clinical trials, and volume of foreign direct investment. In contrast, output indicators such as approval of new chemical entities suggest that the process of catching up has only just commenced. Pharmaceutical innovation is at the stage of mainly generating inputs to integrated processes controlled by the globally incumbent firms. PMID- 19563314 TI - Policy challenges of nanomedicine for Australia's PBS. AB - All major pharmaceutical companies are currently investing significantly in the development of medicines with a nanotechnology component. Such research promises therapeutic drugs with greater efficacy and a wider range of clinical indications. Nanomedicines are just beginning to enter drug regulatory processes, but within a few decades could comprise a dominant group within the class of innovative pharmaceuticals. The current thinking of government safety and cost effectiveness regulators appears to be that these products give rise to few if any nano-specific issues. This article challenges that proposition and seeks to explore what features of nanomedicines may create unique or heightened policy challenges for government systems of cost-effectiveness regulation. The Australian Pharmaceutical Benefits Scheme (PBS) is a key exemplar of the latter type of regulation in that it links expert scientific evaluation of cost effectiveness with the pricing of PBS-listed drugs. In the current global financial crisis such systems are likely to become increasingly attractive and how they handle the demands made upon them by nanomedicines (including by application of a variation of the precautionary principle) is likely to be of considerable interest to policy makers worldwide. PMID- 19563315 TI - Doctor and pharmacist - back to the apothecary! AB - The Australian National Medicines Policy embodies four tenets: availability, quality, safety and efficacy of medicines; timely access to affordable medicines; quality use of medicines (QUM); and a responsible and viable medicines industry. The promotion of QUM requires a multidisciplinary approach, including contributions from government, the pharmaceutical industry, health professionals, consumers and academia. However, there are significant tensions and unintended effects associated with the multidisciplinary approach, especially with the relationships between prescribers and dispensers of medicines. The general practitioner and the pharmacist share a common ancestor - the apothecary. The separation of dispensing from prescribing, which began in medieval Europe and 19th century England, reframed and confined the patient-doctor relationship to one of diagnosis, prescription and non-drug management. The role of pharmacists was limited to dispensing, though the present trend is for their responsibilities to be widened. Historical antecedents, the contribution of an increasing number of actors to the costs of health care, universal health insurance and an evolving regulatory framework, are among the factors influencing doctor-pharmacist relations. The prescribing and dispensing of medicines must be guided by an ethical clinical governance structure encompassing health professionals, regulators, the pharmaceutical industry and consumers. There must be close monitoring of safety and effectiveness, and promotion of quality use of medicines and improved patient outcomes. Ongoing training and professional development, within and across professional boundaries, is essential to support harmonious and cost-effective inter-professional practice. The approach must be "apothecarial" with complementary roles and responsibilities for the prescriber and dispenser within the patient-clinician therapeutic relationship, and not adversarial. PMID- 19563316 TI - A review of proposals to reform the regulation of complementary medicines. AB - In 2003, the Therapeutic Goods Administration instituted a major recall of products made by Pan Pharmaceuticals Limited. Later that year, an expert committee produced 49 recommendations for complementary medicines reform, many of which were to be implemented by the proposed Australia New Zealand Therapeutic Products Authority (ANZTPA). In 2008, the Pan Pharmaceuticals affair reached some conclusion in the courts, the ANZTPA had been abandoned and the case for reform had intensified. There was widespread and increasing use of complementary medicines yet consumers were often unaware that, unlike conventional medicines, these medicines were not evaluated for efficacy. The justification of this two tiered regulatory system was that complementary medicines are relatively low-risk products. However low risk does not mean no risk. A number of consumers have been shown to use these products for conditions where there is no evidence of effect, potentially placing them at risk. In addition, promotion often overstates their benefits while minimising and sometimes denying known adverse effects and drug interactions. Complaint procedures are overloaded and the "sanctions" available do not deter repeat offenders. A number of regulatory reforms have been suggested to overcome these problems; they are reviewed in this paper. PMID- 19563317 TI - Complementary medicines regulatory reform. PMID- 19563320 TI - Models of Care. Editorial. PMID- 19563321 TI - Becoming Australian? Two different approaches to health care reform in the United States. PMID- 19563322 TI - GP access to MRI: the Australian reality. AB - Twelve months ago, the Australian Medical Association (AMA) called upon the Federal Labor Government to implement a previous coalition policy allowing general practitioners to directly refer patients for magnetic resonance imaging (MRI) scans of the knee and brain. To support their position, the AMA commissioned a University of Sydney report evaluating the health care and economic outcomes of the policy. The AMA reported that the results supported the policy and would result in a $42 million saving from fewer computed tomography (CT) scans and fewer specialist referrals and consultations. Arguably, this was not an accurate portrayal of the results. Further research is needed, and ongoing dialogue with radiologists and other key stakeholders is urged, to ensure that access to MRI facilities will continue to meet future demand and that GPs will be adequately trained in utilising MRI services. PMID- 19563323 TI - Working together: collaboration between midwives and doctors in public hospitals. AB - While collaborative, multidisciplinary teamwork is widely espoused as the goal of contemporary hospitals, it is hard to achieve. In maternity care especially, professional rivalries and deep-seated philosophical differences over childbirth generate significant tensions. This article draws on qualitative research in several Victorian public maternity units to consider the challenges to inter professional collaboration. It reports what doctors and midwives looked for in colleagues they liked to work with - the attributes of a "good doctor" or a "good midwife". Although their ideals did not entirely match, both groups respected skill and hard work and sought mutual trust, respect and accountability. Yet effective working together is limited both by tensions over role boundaries and power and by incivility that is intensified by increasing workloads and a fragmented labour force. The skills and qualities that form the basis of "professional courtesy" need to be recognised as essential to good collaborative practice. PMID- 19563324 TI - Cholesterol-lowering therapy and the Australian Pharmaceutical Benefits Scheme: a population study. AB - OBJECTIVE: The Australian Pharmaceutical Benefits Scheme (PBS) expanded the criteria for eligibility for subsidised lipid-lowering therapy (LLT) in 2006. The aim of this study was to determine the use of LLT in a representative Australian population in relation to cardiovascular disease (CVD) risk, and the effectiveness of the therapy in meeting target levels. DESIGN: Cross-sectional biomedical study with telephone interviews, questionnaires, clinical measurements, and PBS dispensing data. SUBJECTS: Representative population sample of 4060 urban adults aged > or = 18 years attending for the biomedical examination in 2001. RESULTS: Of the 406 who qualified for PBS-subsidised LLT at that time, only 88 (21.5%) were actually on LLT. National Heart Foundation of Australia (NHF) recommended low-density lipoprotein cholesterol (LDL-C) levels of < 2.5 mmol/L were recorded in only 13% (528) of the population, and in 46.8% of those on LLT. Of those on LLT, 76% had total cholesterol < 5.5 mmol/L, but over 80% had total cholesterol levels above NHF-recommended levels of 4.0 mmol/L. Of the 842 classified at the highest CVD risk, only 26% were using LLT. Those aged > 60 years and on low incomes were significantly more likely to use LLT. The new PBS criteria will expand eligibility to include nearly 20% of adults. CONCLUSIONS: The majority of people at high risk of CVD were not receiving LLT, and LLT is not being used to its full effectiveness. People with low incomes or on government benefits or pensions were not less likely to use LLT than others under the PBS scheme. Whether higher copayments for those on low incomes who do not qualify for concessional payments is a significant barrier to LLT use needs further research. PMID- 19563325 TI - The AusPSIs: the Australian version of the Agency of Healthcare Research and Quality patient safety indicators. AB - Many countries are seeking ways to measure the safety and performance of their health systems. The ability to track improvement and monitor safety event rates at a population level is provided by routinely collected administrative data in conjunction with a set of well-developed indicators such as the patient safety indicators from the Agency for Healthcare Research and Quality (AHRQ) in the United States of America. These indicators are currently in the International Classification of Diseases Ninth Revision, Clinical Modification (ICD-9-CM) whereas Australia has coded its data in ICD-10-Australian Modification (ICD-10 AM) since 1998. We describe the process recently undertaken to translate and revise the patient safety indicators (PSIs) so they can be of use with ICD-10-AM. The initial translation (electronic mapping, review and revision by expert coder, programming of codes and testing on data from 1996-1998 [ICD 9-CM] to 1998-2006 [ICD-10-AM, through 4 editions]) found that differences between ICD-9-CM and ICD 10-AM datasets presented some challenges. After this phase, which was faithful to AHRQ's case definitions, the indicators were refined for use with the condition onset flag, resulting in the AusPSIs. PMID- 19563326 TI - Live-birth in vipers (Viperidae) is a key innovation and adaptation to global cooling during the Cenozoic. AB - The identification of adaptations and key innovations has long interested biologists because they confer on organisms the ability to exploit previously unavailable ecological resources and respond to novel selective pressures. Although it can be extremely difficult to test for the effects of a character on the rate of lineage diversification, the convergent evolution of a character in multiple lineages provides an excellent opportunity to test for the effect of that character on lineage diversification. Here, I examine the effect of parity mode on the diversification of vipers, which have independently evolved viviparity in at least 13 lineages. I find strong statistical evidence that viviparous species diversify at a greater rate than oviparous species and correlate major decreases in the diversification rate of oviparous species with periods of global cooling, such as the Oligocene. These results suggest that the evolution of viviparity buffered live-bearing species against the negative effects of global climate change during the Cenozoic, and was a key innovation in the evolution and diversification of live-bearing vipers. PMID- 19563327 TI - Mandibular overdentures supported by two Branemark, IMZ or ITI implants: a ten year prospective randomized study. AB - OBJECTIVES: The aim of this prospective comparative study was to evaluate the survival rate, condition of peri-implant tissues, patient satisfaction and surgical and prosthetic aftercare of the IMZ-implant system (two-stage cylinder type), the Branemark-implant system (two-stage screw type) and the ITI-implant system (one-stage screw type) supporting a mandibular overdenture during a 10 year follow-up period. MATERIALS AND METHODS: Three groups of 30 edentulous patients were treated with two endosseous implants in the interforaminal region of the mandible. Clinical and radiographic parameters were evaluated immediately after completion of the prosthetic treatment and after 1, 5 and 10 years of functional loading. Prosthetic and surgical aftercare was scored during the evaluation period, as well as patient satisfaction. RESULTS: The 10-year survival rate was 93% for the IMZ group, 98% for the Branemark group and 100% for the ITI group (IMZ6 mm at baseline and 3 months in 25 periodontitis subjects and two sites in 25 periodontally healthy subjects. Samples were analysed for their content of 13 bacterial species using both assays. Similarities for each species between techniques were determined using regression analysis. Differences between health and periodontitis were determined using the Mann-Whitney test. RESULTS: Three hundred and fifty samples were evaluated using both techniques. Regression analysis indicated that 10/13 test species showed significant positive correlations between the counts determined by checkerboard analysis and levels determined by the PCR-based test after adjusting for 13 comparisons. The highest rank correlations of 0.58, 0.49 and 0.46 were seen for Treponema denticola, Fusobacterium nucleatum and Eubacterium nodatum, respectively (p<0.0001). Both tests could distinguish samples from healthy and periodontitis subjects. CONCLUSION: Detection patterns of 10/13 test species in subgingival plaque samples from periodontitis and healthy subjects were similar using the two molecular techniques. PMID- 19563331 TI - One-year outcomes of repeated adjunctive photodynamic therapy during periodontal maintenance: a proof-of-principle randomized-controlled clinical trial. AB - BACKGROUND: Single photodynamic therapy (PDT) has been effective in initial periodontal therapy, but only improved bleeding on probing (BoP) in maintenance patients after a single use. Repeated PDT has not been addressed. OBJECTIVES: To study the possible added benefits of repeated adjunctive PDT to conventional treatment of residual pockets in patients enrolled in periodontal maintenance. MATERIAL AND METHODS: Ten maintenance patients with 70 residual pockets [probing pocket depth (PPD)>or=5 mm] were randomly assigned for treatment five times in 2 weeks (Days 0, 1, 2, 7, 14) with PDT (test) or non-activated laser (control) following debridement. The primary outcome variable was PPD, and the secondary variables were clinical attachment level (CAL) and BoP. These were assessed at 3, 6 and 12 months following the interventions. RESULTS: Greater PPD reductions were observed in the test (-0.67 +/- 0.34; p=0.01) compared with the control patients (-0.04 +/- 0.33; NS) after 6 months. Significant CAL gain (+0.52 +/- 0.31; p=0.01) was noted for the test, but not in the control (-0.27 +/- 0.52; NS) patients after 6 months. BoP percentages decreased significantly in test (97-64%, 67%, 77%), but not control patients after 3, 6 and 12 months. CONCLUSIONS: Repeated (five times) PDT adjunctive to debridement yielded improved clinical outcomes in residual pockets in maintenance patients. The effects were best documented after 6 months. PMID- 19563332 TI - Long-term management of the liver transplant patient: recommendations for the primary care doctor. AB - No official document has been published for primary care physicians regarding the management of liver transplant patients. With no official source of reference, primary care physicians often question their care of these patients. The following guidelines have been approved by the American Society of Transplantation and represent the position of the association. The data presented are based on formal review and analysis of published literature in the field and the clinical experience of the authors. These guidelines address drug interactions and side effects of immunosuppressive agents, allograft dysfunction, renal dysfunction, metabolic disorders, preventive medicine, malignancies, disability and productivity in the workforce, issues specific to pregnancy and sexual function, and pediatric patient concerns. These guidelines are intended to provide a bridge between transplant centers and primary care physicians in the long-term management of the liver transplant patient. PMID- 19563333 TI - Mumps: not an innocent bystander in solid organ transplantation. AB - Recently two major outbreaks of mumps have occurred: in the UK more than 56,000 cases were notified between 2004 and 2005, and in the United States, 6,584 cases were reported in 2006. Most patients were young healthy adults, in whom mumps normally has a benign course. Little is known about mumps in the immunocompromised patient. Here, we report a case of a 56-year renal transplant recipient who developed acute irreversible transplant failure due to interstitial nephritis caused by mumps. RNA of the mumps virus was detected in the urine as well as in a renal biopsy. In view of the ongoing presence of the mumps virus in the population, one should be aware of the possible occurrence of this infection in immunocompromised patients. PMID- 19563334 TI - Clinical implications of advances in the basic science of liver repair and regeneration. AB - Recent advances in our understanding of the basic mechanisms that control liver regeneration and repair will produce the next generation of therapies for human liver disease. Insights gained from large-scale genetic analysis are producing a new framework within which to plan interventions. Identification of specific molecules that drive regeneration will increase the options for live-donor liver transplantation, and help treat patients with small-for-size syndrome or large tumors who would otherwise have inadequate residual mass after resection. In a complementary fashion, breakthroughs in the ability to manipulate various cell types to adopt the hepatocyte or cholangiocyte phenotype promise to revolutionize therapy for acute liver failure and metabolic liver disease. Finally, elucidating the complex interactions of liver cells with each other and various matrix components during the response to injury is essential for fabricating a liver replacement device. This focused review will discuss how a variety of important scientific advances are likely to impact the treatment of specific types of liver disease. PMID- 19563335 TI - Asynchronous, out-of-sequence, transcontinental chain kidney transplantation: a novel concept. AB - The organ donor shortage has been the most important hindrance in getting listed patients transplanted. Living kidney donors who are incompatible with their intended recipients are an untapped resource for expanding the donor pool through participation in transplant exchanges. Chain transplantation takes this concept further, with the potential to benefit even more recipients. We describe the first asynchronous, out of sequence transplant chain that was initiated by transcontinental shipment of an altruistic donor kidney 1 week after that recipient's incompatible donor had already donated his kidney to the next recipient in the chain. The altruistic donor kidney was transported from New York to Los Angeles and functioned immediately after transplantation. Our modified sequence asynchronous transplant chain (MATCH) enabled eight recipients, at four different institutions, to benefit from the generosity of one altruistic donor and warrants further exploration as a promising step toward addressing the organ donor shortage. PMID- 19563336 TI - Screening to prevent polyoma virus nephropathy in kidney transplantation: a cost analysis. AB - Polyoma virus nephropathy is an important cause of graft dysfunction in kidney transplant recipients and screening to prevent disease has been advocated. Although screening incurs new costs, our hypothesis is that savings from less immunosuppression in those with positive screening tests could pay for overall costs of screening. In 134 consecutive recipients, polyoma virus (positive decoy cells) was detected in the urine of 34 (25.4%) individuals over a 2-year follow up. Of these 34, 11 had a plasma BK PCR of >7700 copies/mL. Immunosuppression was reduced stepwise in these patients until viral loads fell <1000/mL. Overall screening costs (including extra plasma PCR testing) were estimated at $33,450. Those with positive PCR had greater reductions in annual immunosuppression costs by year 2 ($6452 vs. $2799, p = 0.0015) compared to those with negative screens. At the end of the 2-year period, 61% of the screening costs were covered by less immunosuppressant costs. At the end of 30 months there were net savings. In summary, reductions in immunosuppression cover the cost of screening for polyoma viral infection. Longer-term follow-up is needed to ensure patient outcomes remain acceptable. PMID- 19563337 TI - Cancer mortality in kidney transplantation. AB - Immunosuppression is associated with an increased risk of cancer in kidney transplant recipients compared to the general population. It is less clear whether standardized cancer mortality ratios (SMRs) are also increased. This study's hypothesis is that SMRs are not increased because of competing risks of death. During the median follow-up of 5.05 years (Q1-Q3: 2.36-8.62), there were 1937 cancer deaths and 36 619 noncancer deaths among 164 078 first kidney-only transplant recipients captured in the United States Renal Data System between January 1990 and December 2004. The observed cancer death rate was 206 per 100 000 patient-years compared to an expected rate of 215 per 100,000 patient-years in the general population. The overall age- and sex-adjusted SMR was only 0.96 (95% CI 0.92-1.00). However, patients <50 years had SMRs significantly greater than unity while patients >60 had SMRs lower than unity. Up to 25% of cancer related deaths occurred after allograft failure. These findings challenge the notion that cancer is a major cause of premature death in all kidney transplant recipients and has implications for design of cancer prevention strategies in kidney transplant recipients. PMID- 19563338 TI - Scoring total inflammation is superior to the current Banff inflammation score in predicting outcome and the degree of molecular disturbance in renal allografts. AB - Emerging molecular analysis can be used as an objective and independent assessment of histopathological scoring systems. We compared the existing Banff i score to the total inflammation (total i-) score for assessing the molecular phenotype in 129 renal allograft biopsies for cause. The total i-score showed stronger correlations with microarray-based gene sets representing major biological processes during allograft rejection. Receiver operating characteristic curves showed that total-i was superior (areas under the curves 0.85 vs. 0.73 for Banff i-score, p = 0.012) at assessing an abnormal cytotoxic T cell burden, because it identified molecular disturbances in biopsies with advanced scarring. The total-i score was also a better predictor of graft survival than the Banff i-score and essentially all current diagnostic Banff categories. The exception was antibody-mediated rejection which is able to predict graft loss with greater specificity (96%) but at low sensitivity (38%) due to the fact that it only applies to cases with this diagnosis. The total i score is able to achieve moderate sensitivities (60-80%) with losses in specificity (60-80%) across the whole population. Thus, the total i-score is superior to the current Banff i-score and most diagnostic Banff categories in predicting outcome and assessing the molecular phenotype of renal allografts. PMID- 19563339 TI - Calcineurin inhibitor minimization in the Symphony study: observational results 3 years after transplantation. AB - The Symphony study showed that at 1 year posttransplant, a regimen based on daclizumab induction, 2 g mycophenolate mofetil (MMF), low-dose tacrolimus and steroids resulted in better renal function and lower acute rejection and graft loss rates compared with three other regimens: two with low-doses of cyclosporine or sirolimus instead of tacrolimus and one with no induction and standard cyclosporine dosage. This is an observational follow-up for 2 additional years with the same endpoints as the core study. Overall, 958 patients participated in the follow-up. During the study, many patients changed their immunosuppressive regimen (e.g. switched from sirolimus to tacrolimus), but the vast majority (95%) remained on MMF. During the follow-up, renal function remained stable (mean change: -0.6 ml/min), and rates of death, graft loss and acute rejection were low (all about 1% per year). The MMF and low-dose tacrolimus arm continued to have the highest GFR (68.6 +/- 23.8 ml/min vs. 65.9 +/- 26.2 ml/min in the standard dose cyclosporine, 64.0 +/- 23.1 ml/min in the low-dose cyclosporine and 65.3 +/- 26.2 ml/min in the low-dose sirolimus arm), but the difference with the other arms was not significant (p = 0.17 in an overall test and 0.077, 0.039 and 0.11, respectively, in pair-wise tests). The MMF and low-dose tacrolimus arm also had the highest graft survival rate, but with reduced differences between groups over time, and the least acute rejection rate. In the Symphony study, the largest ever prospective study in de novo kidney transplantation, over 3 years, daclizumab induction, MMF, steroids and low-dose tacrolimus proved highly efficacious, without the negative effects on renal function commonly reported for standard CNI regimens. PMID- 19563340 TI - High weight differences between donor and recipient affect early kidney graft function--a role for enhanced IL-6 signaling. AB - The frequency of delayed function of kidney transplants varies greatly and is associated with quality of graft, donor age and the duration of cold ischemia time. Furthermore, body weight differences between donor and recipient can affect primary graft function, but the underlying mechanism is poorly understood. We transplanted kidney grafts from commensurate body weight (L-WD) or reduced body weight (H-WD) donor rats into syngeneic or allogeneic recipients. Twenty-four hours posttransplantation, serum creatinine levels in H-WD recipients were significantly higher compared to L-WD recipients indicating impaired primary graft function. This was accompanied by upregulation of IL-6 transcription and increased tubular destruction in grafts from H-WD recipients. Using DNA microarray analysis, we detected decreased expression of genes associated with kidney function and an upregulation of other genes such as Cyp3a13, FosL and Trib3. A single application of IL-6 into L-WD recipients is sufficient to impair primary graft function and cause tubular damage, whereas immediate neutralization of IL-6 receptor signaling in H-WD recipients rescued primary graft function with well-preserved kidney graft architecture and a normalized gene expression profile. These findings have strong clinical implication as anti-IL6R treatment of patients receiving grafts from lower-weight donors could be used to improve primary graft function. PMID- 19563341 TI - Perception versus reality?: Virtual crossmatch--how to overcome some of the technical and logistic limitations. AB - The goal of this work was to evaluate concordance between (a) actual flow cytometric crossmatch (FCXM) that is performed by the OPO laboratory servicing our transplant center and (b) virtual XM (vXM) prediction based on antibody identification by solid-phase methods performed in our laboratory. A total of 1586 FCXM, performed between June 2007 and September 2008, between all potential deceased donors in our region and sera from patients awaiting kidney or kidney pancreas transplant, listed at Northwestern Memorial Hospital were evaluated. A key finding of this analysis was the understanding that a thorough vXM cannot be performed in some donor/recipient pairs due to the lack of certain antibody profile data specific to the donor in question. Obtaining more in depth and stringent information regarding antibody specificities, we demonstrate an excellent sensitivity and specificity of the vXM assays- 86.1% and 96.8%, respectively, with a positive likelihood ratio and negative likelihood ratios of 26.9 and 0.14, respectively. The vXM can serve as an outstanding tool to predict HLA compatibility between donor and recipient, with the caveat that the presence/absence of all antibodies against the potential donor and their strength have been thoroughly investigated. PMID- 19563342 TI - Primed CD8(+) T-cell responses to allogeneic endothelial cells are controlled by local complement activation. AB - CD8 T cells primed by transplantation recognize allogeneic class I MHC molecules expressed on graft vascular endothelium and contribute to allograft injury. We previously showed that immune cell-derived complement activation fragments are integral to T cell activation/expansion. Herein we tested the impact of local complement production/activation on T cell/endothelial cell (EC) interactions. We found that proinflammatory cytokines upregulated alternative pathway complement production by ECs, yielding C5a. We further found that ECs deficient in the cell surface C3/C5 convertase regulator decay accelerating factor (DAF, CD55) induced greater CD8 T-cell proliferation and more IFNgamma(+) and perforin(+) effector cells than wild-type (WT) ECs. Allogeneic C3(-/-) EC induced little or no CD8 responses. Abrogation of responses following C5a receptor (C5aR) blockade, or augmentation following addition of recombinant C5a demonstrated that the effects were mediated through T-cell-expressed-C5aR interactions. Analyses of in vivo CD8 cell responses to transplanted heart grafts deficient in EC DAF showed similar augmentation. The findings reveal that EC-derived complement triggers secondary CD8 T-cell differentiation and expansion and argue that targeting complement and/or C5aR could limit T-cell-mediated graft injury. PMID- 19563343 TI - Lack of effect of MICA antibodies on graft survival following heart transplantation. AB - Little is known about the effect of MICA antibodies (Abs) on cardiac allograft function and survival. Pretransplant and posttransplant serum from 491 and 196 adult cardiac allograft recipients, respectively, has been investigated for MICA Abs, donor specificity and the effect of MICA Abs on graft survival, acute rejection episodes (AR) and cardiac allograft vasculopathy (CAV). Patients with HLA Abs (11.6%) were excluded from the analysis. A total of 11.8% of patients had MICA Abs, without HLA Abs, before their transplant. Actuarial graft survival demonstrated slightly better survival of patients with donor-specific MICA Abs at 1 and 5 years (88.9% and 83.3%) than patients negative for MICA Abs (72% and 63.7%, p = 0.051). After transplantation, 15.8% of patients produced MICA Abs, and in 17 patients these were produced de novo. There was no effect of pretransplant or posttransplant production of MICA Abs on numbers of AR episodes in year 1, or CAV assessed at years 3 and 5. Immunocytochemistry of cardiac biopsies from 11 patients did not demonstrate a presence of MICA. Sera from only 4/69 patients with MICA Abs fixed complement prior to transplantation and from 7/38 patients following transplantation. In conclusion, this study suggests that MICA Abs do not adversely affect the outcome of cardiac transplantation. PMID- 19563344 TI - Infusion of mesenchymal stem cells and rapamycin synergize to attenuate alloimmune responses and promote cardiac allograft tolerance. AB - The inherent immunosuppressive properties and low immunogenicity of mesenchymal stems cells (MSCs) suggested their therapeutic potential in transplantation. We investigated whether MSCs could prolong allograft survival. Treatment involving infusion of MSCs into BALB/c recipients 24 hours after receiving a heart allograft from a C57BL/6 donor significantly abated rejection and doubled graft mean survival time compared to untreated recipients. Furthermore, combination therapy of MSCs and low-dose Rapamycin (Rapa) achieved long-term heart graft survival (>100 days) with normal histology. The treated recipients readily accepted donor skin grafts but rejected third-party skin grafts, indicating the establishment of tolerance. Tolerant recipients exhibited neither intragraft nor circulating antidonor antibodies, but demonstrated significantly high frequencies of both tolerogenic dendritic cells (Tol-DCs) and CD4(+)CD25(+)Foxp3(+)T cells in the spleens. Infusion of GFP(+)C57BL/6-MSCs in combination with Rapa revealed that the GFP-MSCs accumulated in the lymphoid organs and grafts of tolerant recipients. Thus, engraftment of infused MSCs within the recipient's lymphoid organs and allograft appeared to be instrumental in the induction of allograft specific tolerance when administered in combination with a subtherapeutic dose of Rapamycin. This study supports the clinical applicability of MSCs in transplantation. PMID- 19563345 TI - Long-term survival in patients with status epilepticus: a tertiary referral center study. AB - PURPOSE: To determine long-term survival in patients with status epilepticus (SE). METHODS: We prospectively followed patients admitted for the first (69.6%) or recursive episode of SE between January 1, 1989 and December 31, 1997 at the Institute of Neurology, Belgrade, Serbia, until death or study termination (December 31, 2006). Data were obtained for cause of death; etiology of SE-acute symptomatic (AS), progressive symptomatic (PS), remote symptomatic (RS), and idiopathic/cryptogenic (I/C); presence of epilepsy; and reoccurrence of SE. Standardized mortality rate (SMR), survival, and regression analysis were used. RESULTS: A total of 120 of 750 patients with an episode of SE (15.9%) died in the 30-day period following SE. Data for 207 of 630 (32.8%) surviving patients (35.7% with initial SE) were available at the end of follow-up [median 12 years; 95% confidence interval (CI) 11.1-12.8]. SMR was significantly increased (SMR = 1.81; 95% CI 1.32-2.41). There were 46 deaths (22.2%): 15 of 65 in the AS, 20 of 29 in the PS, 6 of 29 in the RS, and 5 of 75 in the I/C groups. Five-year survival rate was lowest in the PS (45%) compared to AS (91%), RS (87%), and I/C (99%) groups. The following characteristics increased long-term risk for mortality: older age [Exp(B) 1.05, 95% CI 1.029-1.072], PS and AS etiology [Exp(B) 15.6, 95% CI 5.8 41.6; 3.3, 95% CI 1.2-9.1], presence of epilepsy [Exp(B) 2.3, 95% CI 1.2-4.3], and initial SE [Exp(B) 2.4, 95% CI 1.4-4.4]. DISCUSSION: Approximately one of five patients die within 12 years after an episode of SE. Symptomatic SE (PS and AS), initial SE, age, and presence of epilepsy are associated with long-term increased risk of death. PMID- 19563346 TI - Long-term use of felbamate: clinical outcomes and effect of age and concomitant antiepileptic drug use on its clearance. AB - PURPOSE: To determine the effects of long-term use of felbamate (FBM) on weight, complete blood count, liver function tests, and seizure control, and also to determine the effect of age on FBM clearance. METHODS: A computerized prospective database was used to identify all subjects who had FBM listed as one of their antiepileptic drugs (AEDs) during their most recent clinic visit. Medical records from each patient were then reviewed for inclusion criteria [treatment >2 years, FBM initiated at the study clinic, data for pre-FBM (Time-1; T1), one-year exposure to FBM (Time-2; T2), and the latest visit (Time-3; T3)]. Clinical information was abstracted from clinic charts. RESULTS: Seventy-seven patients (F = 41, M = 36; ages 10-69 years) met entry criteria. Mean treatment time was 7.4 years, with the longest 20.3 years. Significant weight loss (mean 5.1 kg; p < 0.001) occurred from T1 to T2, but weight was regained by T3. No clinically significant changes in laboratory parameters occurred. Older age was associated with a significant decrease in FBM clearance (p = 0.01). Significant reduction in generalized tonic-clonic seizures was seen at both T2 (p < 0.001) and T3 (p < 0.001) compared with seizure frequency at T1. DISCUSSION: Our results suggest that long-term FBM use is associated with persistent decrease of seizures and no clinically significant changes in major laboratory parameters. Older age correlated with reduced apparent clearance of FBM. The patient population described in this study were long-term users of FBM who were continuing to use the drug, and thus this study does not constitute an "intent to treat" study. PMID- 19563347 TI - High seizure frequency prior to antiepileptic treatment is a predictor of pharmacoresistant epilepsy in a rat model of temporal lobe epilepsy. AB - PURPOSE: Progress in the management of patients with medically intractable epilepsy is impeded because we do not fully understand why pharmacoresistance happens and how it can be predicted. The presence of multiple seizures prior to medical treatment has been suggested as a potential predictor of poor outcome. In the present study, we used an animal model of temporal lobe epilepsy to investigate whether pharmacoresistant rats differ in seizure frequency from pharmacoresponsive animals. METHODS: Epilepsy with spontaneous recurrent seizures (SRS) was induced by status epilepticus. Frequency of SRS was determined by video/EEG (electroencephalography) monitoring in a total of 33 epileptic rats before onset of treatment with phenobarbital (PB). RESULTS: Thirteen (39%) rats did not respond to treatment with PB. Before treatment with PB, average seizure frequency in PB nonresponders was significantly higher than seizure frequency in responders, which, however, was due to six nonresponders that exhibited > 3 seizures per day. Such high seizure frequency was not observed in responders, demonstrating that high seizure frequency predicts pharmacoresistance in this model, but does not occur in all nonresponders. DISCUSSION: The data from this study are in line with clinical experience that the frequency of seizures in the early phase of epilepsy is a dominant risk factor that predicts refractoriness. However, resistance to treatment also occurred in rats that did not differ in seizure frequency from responders, indicating that disease severity alone is not sufficient to explain antiepileptic drug (AED) resistance. These data provide further evidence that epilepsy models are useful in the search for predictors and mechanisms of pharmacoresistance. PMID- 19563348 TI - Long-term efficacy and tolerability of topiramate as add-on therapy in refractory partial epilepsy: an observational study. AB - PURPOSE: To evaluate the long-term efficacy and tolerability of topiramate (TPM) as add-on therapy in patients with refractory partial epilepsy. METHODS: This is a retrospective, single-center, long-term observational study. Patients fulfilling the criteria of medical intractability proposed by Berg et al. were entered into the study if they were newly prescribed TPM as add-on therapy between January 2000 and June 2002. The usual starting dosage of TPM was 50 mg/day and optimal-dose adjustments were made according to individual clinical responses. Efficacy and tolerability were analyzed every year during 5-year follow-up in the "intention-to-treat (ITT) population." Retention rate was estimated by Kaplan-Meyer analysis. RESULTS: A total of 125 patients were included in the study and 107 patients (85.6%) were followed for 5 years. Retention rate was 87.2% at 1 year and 64% at 5 years. At the end of 5 years, the median seizure frequency reduction rate was 69.0% and responder rate was 43.2% in the ITT population. Cumulative seizure-free rate (SFR) was 30.4% and the terminal 1-year SFR was 12.8% in the ITT population (20.0% in completers) at 5-year follow up. Adverse events (AEs) occurred in 39.2% of patients, including significant AEs leading to antiepileptic drug (AED) withdrawal in 14.4%. The most common AEs were anorexia (16.0%), weight loss (10.4%), and gastrointestinal symptoms (8.8%). Concomitant AEDs were reduced in 25.0% of the completers. DISCUSSION: Low-dose and slow-dose escalation of TPM in add-on therapy for patients with refractory partial epilepsy is effective and well tolerated in long-term, individualized clinical practice. PMID- 19563349 TI - Hemiconvulsion-hemiplegia syndrome in a patient with severe myoclonic epilepsy in infancy. AB - We report a 2-year-old girl who had repeated febrile or afebrile seizures since infancy. Prolonged left/right hemiconvulsions and myoclonus of the eyelids/extremities with generalization to tonic-clonic seizures, were refractory to antiepileptic agents. At age 1 year and 4 months, she contracted rotavirus infection, and developed status epilepticus with persistent right hemiclonic seizures. Left unilateral brain edema with subsequent emergence of cortical laminar necrosis and white matter lesions, and progressive atrophy of the left cerebral hemisphere were noted during this period. She showed residual right hemiparesis and mild intellectual disability, and had generalized/eyelid myoclonia and hot water epilepsy after a 5-month seizure-free period. Analysis for SCN1A, the gene encoding the neuronal voltage-gated Na+ channel alpha1 subunit revealed a nonsense mutation, R1892X. These indicate the potential risk in patients with severe myoclonic epilepsy in infancy (SMEI) to develop hemiconvulsion-hemiplegia (HH) syndrome. SCN1A mutations may need to be further explored in patients with HH syndrome without features of SMEI. PMID- 19563350 TI - Natural history and mortality of chronic epilepsy in an untreated population of rural Bolivia: a follow-up after 10 years. AB - PURPOSE: To evaluate the natural history and mortality of chronic epilepsy in an untreated prevalence cohort of people with epilepsy (PWE) in a rural area of Bolivia. METHODS: During 1994-1996 we carried out an epidemiologic survey in a sample of 9,995 subjects in the Cordillera province. At the end of the survey we identified 130 PWE, of whom 118 were classified as having "active epilepsy." We revisited this cohort 10 years after the prevalence survey. RESULTS: We were able to trace 103 (87.3%) of the 118 PWE previously identified. Ten of the 103 subjects died during the follow-up period. Of the 93 PWE still alive, adequate information on the occurrence of seizures was available for 71 subjects, of whom 31 (43.7%) were seizure-free for more than 5 years; only 3 of these 31 subjects have taken an antiepileptic drug (AED) for more than 1 year. Generalized seizures were associated with a better prognosis. Mortality rate in our prevalent cohort was 10.0/1,000 person-year at risk [95% confidence interval (CI) 5.5-18.3], without a significant increased risk respect to the general population [standardized mortality rate (SMR) 1.34; 95% CI 0.68-2.39]; a significant increased risk of death was found for patients with remote symptomatic epilepsy (SMR 3.0; 95% CI 1.2-6.3) but not with idiopathic epilepsy. Three of the 10 subjects died of causes possibly related to epilepsy. DISCUSSION: Our data suggest that spontaneous remission of epilepsy occurs in a substantial proportion of untreated patients affected by chronic epilepsy; concerning mortality, we found a 3-fold increased mortality in patients with remote symptomatic epilepsy. PMID- 19563351 TI - Cardiac calsequestrin: the new kid on the block in arrhythmias. AB - Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a rare inherited disease characterized by physical or emotional stress-induced ventricular arrhythmias in the absence of any structural heart disease or QT prolongation. Thus far, mutations in genes encoding the sarcoplasmic reticulum Ca(2+) release channel (RYR2) and the sarcoplasmic reticulum Ca(2+) binding protein cardiac calsequestrin (CASQ2) have been identified in CPVT patients. Here, we review the role of cardiac calsequestrin in health and disease, with a particular focus on how calsequestrin deficiency can cause arrhythmia susceptibility. Clinical implications and a promising new drug therapy for CPVT are discussed. PMID- 19563352 TI - Toys R Us: do they truly work? PMID- 19563353 TI - Predicting outcome from AF ablation: size of the chamber, or is tissue the issue? PMID- 19563354 TI - Differences between women and men with ICDs: myth or reality? PMID- 19563355 TI - Is the isolation of the pulmonary vein completed? PMID- 19563356 TI - Effective management of ICD patient psychosocial issues and patient critical events. AB - The clinical management of implantable cardioverter-defibrillator (ICD) patients involves successful medical and psychosocial care to reduce mortality and morbidity. Desirable quality of life (QoL) and psychosocial outcomes for ICD patients are achievable for a majority of ICD patients. Patient critical events, such as ICD shocks or ICD recalls, may occur that can dramatically alter the course of patient adjustment if not properly managed. Continuing care strategies that attend to patient critical events as they emerge may improve the psychosocial adjustment and improve the return to optimal daily functioning for ICD patients. This paper reviews QoL and psychosocial outcomes for ICD patients, patient critical events, and clinical implications for patient care. Patient critical events discussed in this paper include perioperative education, ICD shock events, device recalls, and end of life. The clinical management strategies for each of these patient critical events are suggested including patient education, psychosocial information provision, activity prescriptions, recall planning, and shock planning. PMID- 19563357 TI - Association of transseptal punctures with isolated migraine aura in patients undergoing catheter ablation of cardiac arrhythmias. AB - BACKGROUND: Transseptal catheterization (TSC) is performed during catheter ablation involving the lefthand side of the heart. TSC causes a transient iatrogenic atrial septal defect that can predispose patients to migraine episodes. However, isolated migraine aura episodes in patients undergoing TSC have not been described. METHODS: Five hundred seventy-one procedures involving TSC were performed over a 3-year duration. Of these, 3 patients presented with visual symptoms in the first month after the procedure. One patient underwent a TSC during catheter ablation of left-sided accessory pathway and 2 patients underwent TSC during catheter ablation of atrial fibrillation. RESULTS: The incidence of migraine aura in this patient population was 0.5%. In the first week after the procedure, all 3 patients experienced transient reversible visual symptoms of scintillating scotoma consistent with migraine aura. None of the patients had an associated headache. The workup for stroke or transient ischemic attack was negative. All the patients recovered completely within 1 hour of symptom onset and did not have any sequelae. CONCLUSION: This study reports for the first time the incidence and outcomes of isolated migraine aura in patients undergoing electrophysiology procedures involving TSC. For post-TSC patients who present with atypical neurologic symptoms, especially "scintillating scotoma," once transient ischemic attack or other neurologic event has been ruled out, an aura associated with the TSC should be entertained as a possible diagnosis. Electrophysiologists who perform TSC, need to be aware of this phenomenon and can reassure the patients of the transient and benign nature. PMID- 19563358 TI - Cryoballoon pulmonary vein isolation guided by transesophageal echocardiography: novel aspects on an emerging ablation technique. AB - BACKGROUND: Pulmonary vein (PV) isolation using a balloon-mounted cryoablation system is a new technology for the percutaneous treatment of atrial fibrillation (AF). Transesophageal echocardiography (TEE) allows real-time visualization of cryoballoon positioning and successful vein occlusion via color Doppler. We hypothesized that PV mechanical occlusion monitored with TEE could predict effective electrical isolation. METHODS: We studied 124 PVs in 30 patients. Under continuous TEE assessment, a cryoballoon was placed in the antrum of each PV aiming for complete PV occlusion as documented by color Doppler. At the end of the procedure, PV electrical isolation was evaluated using a circumferential mapping catheter. RESULTS: Of the 124 PVs studied, 123 (99.2%) could be visualized by TEE: the antrum was completely visualized in 80 of them (64.5%), partially in 36 (29.0%), and only disappearance of proximal flow could be observed in the remaining 7 PVs (5.7%). Vein occlusion could be achieved in 111 of the 123 (90.2%) visualized PVs. Postinterventional mapping demonstrated electrical isolation in 109 of 111 occluded PVs (positive predictive value 98.2%) and only in 1 of 12 nonoccluded PVs (negative predictive value 91.7%, P < 0.001). After a mean follow-up of 7.4 +/- 3.7 months, 73.3% of patients remained in sinus rhythm without antiarrhythmic drugs. CONCLUSION: Color Doppler documented PV occlusion during cryoballoon ablation can predict effective electrical isolation. PMID- 19563360 TI - Electrophysiologic testing for risk stratification following myocardial infarction: dead or alive? PMID- 19563359 TI - Manifestation of left atrial events and interatrial frequency gradients in the surface electrocardiogram during atrial fibrillation: contributions from posterior leads. AB - BACKGROUND: In most patients, atrial fibrillation (AF) is initiated and maintained by pulmonary vein foci, but the relationship between left atrial (LA) events and the surface electrocardiogram (ECG) is largely unknown. We investigated whether LA events are reflected in the surface ECG and whether additional information can be obtained from recording posterior leads in patients with AF. METHODS AND RESULTS: In 10 patients undergoing radiofrequency ablation of AF, we identified 103 5-second segments with a significant frequency gradient between right (RA) and left (LA) intraatrial electrograms, or with frequency changes from segment to segment in the same patient. QRS-T cancellation methods were used to isolate atrial activity in the surface ECG and peak frequencies were computed. Peak frequencies of different posterior leads were very similar (6.0 +/ 1.3 Hz for V10, 6.0 +/- 0.9 Hz for V9, 5.9 +/- 1.4 Hz for V8, 6.0 +/- 1.3 Hz for V7). We found a strong correlation between V1 and RA and between V9 and LA, 0.89 and 0.88, respectively, while the lowest correlation was found between lead V1 and LA, 0.62, P < 0.0001. Magnitude-squared coherence values were highest between V1 and RA and between V9 and LA. CONCLUSION: We have demonstrated that, by recording additional surface ECG leads from posterior locations, RA and LA electrical events and interatrial frequency gradients can be monitored noninvasively. PMID- 19563361 TI - Electroanatomical endocardial mapping of the specialized conduction system in corrected transposition of the great arteries. PMID- 19563362 TI - Sotalol and a broken heart. AB - An 82-year-old woman with persistent atrial fibrillation underwent successful electrical cardioversion and was begun on sotalol. After 3 days of in-hospital observation she had only mild lengthening of the QT interval. Two weeks later in clinic, the day after her husband's unexpected death, she was noted to have profound QT interval prolongation. Although she was asymptomatic and echocardiography did not disclose regional wall motion abnormalities consistent with takotsubo cardiomyopathy, she probably had a forme fruste of stress cardiomyopathy. Following emotional trauma, a period of heightened vigilance for ventricular proarrhythmia is probably warranted in women treated with antiarrhythmic drugs that lengthen repolarization. PMID- 19563363 TI - Activation sequence change during left free wall pathway ablation: what is the mechanism? PMID- 19563365 TI - Comparison of cooled-tip versus 4-mm-tip catheter in the efficacy of acute ablative tissue injury during circumferential pulmonary vein isolation. AB - INTRODUCTION: Although several studies have reported the benefits of cooled-tip ablation for circumferential pulmonary veins isolation (CPVI), the acute change of substrate property and acute PV reconnection have not been well demonstrated. The aim of this study was to compare the cooled-tip with regular 4-mm-tip catheter in acute substrate change after CPVI and long-term efficacy. METHODS AND RESULTS: One hundred and fifty-six patients (115 males, age 53 +/- 12 years) who underwent CPVI for treatment of atrial fibrillation (AF) were included. Group A consisted of 52 patients with cooled-tip ablation, and group B consisted of 104 patients with 4-mm-tip catheter ablation. The bipolar voltage of circumferential lesions was obtained using a 3-dimensional (3D) mapping system (NavX) before and after CPVI. The electrical reconnections of 4 PVs were evaluated 30 minutes after CPVI using a circular catheter. Cooled-tip catheter caused more reduction of the electrical voltage in PV antrum, lower incidence of acute PV reconnection, inducibility of AF, and gap-related atrial tachyarrhythmia (AT). Less number of left atrial (LA) ablation line and ablation applications and less procedure time were found in cooled-tip group compared to 4-mm-tip group. No significant difference in the incidence of pain sensation and complication was observed between the 2 groups. At a 14-month follow-up, the recurrence rate in the cooled tip group was lower than in the 4-mm group (13.5% vs 33.7%, P = 0.009). CONCLUSION: Cooled-tip catheter has a superior long-term outcome than the 4-mm tip catheter in CPVI, which may be associated with the efficacy of transmural block and electrical isolation in PV antrum. PMID- 19563364 TI - Effect of metoprolol on quality of life in the Prevention of Syncope Trial. AB - INTRODUCTION: Vasovagal syncope is common, often recurrent, and reduces quality of life. No therapies have proven useful to improve quality of life in adequately designed randomized clinical trials. Beta-blockers have mixed evidence for effectiveness in preventing syncope. METHODS: The Prevention of Syncope Trial was a randomized, placebo-controlled, double-blind, multinational, clinical trial that tested the hypothesis that metoprolol improves quality of life in adult patients with vasovagal syncope in a 1-year observation period. Randomization was stratified in strata of patients <42 and > or =42 years old. The quality of life questionnaires Short Form-36 (SF-36) and Euroqol EQ-5D were completed at baseline and after 6 and 12 months of treatment by 204, 132, and 121 patients, respectively. RESULTS: There were 208 patients, mean age 42 +/- 18, of whom 134 (64%) were females. All had positive tilt tests. There was no improvement in quality of life during the trial in the entire group or in either treatment arm. Patients in the metoprolol treatment arm did not have improved quality of life compared to the patients in the placebo arm using either the SF-36 or EQ5D after either 6 or 12 months. Finally, there was no improvement in quality of life associated with metoprolol use in patients either <42 or > or =42 years of age. CONCLUSION: Metoprolol does not improve quality of life in patients with recurrent vasovagal syncope and a positive tilt test. PMID- 19563366 TI - Patients with scar-related right ventricular tachycardia: determinants of long term outcome. AB - INTRODUCTION: Patients with established arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVC/D) based on task force (TF) criteria and ventricular tachycardia (VT) are at risk of VT recurrence and sudden death. Data on patients with VT due to right ventricular (RV) scar not fulfilling TF criteria are lacking. The purpose of this study was to assess the long-term arrhythmia recurrence rate and outcome in patients with scar-related right VT with and without a diagnosis of ARVC/D. METHODS: Sixty-four patients (age 43.5 +/- 15 years, 49 males) presenting with nonischemic scar-related VT of RV origin were studied. Scar was identified by electroanatomical mapping, contrast echocardiography, and/or magnetic resonance imaging (MRI). Patients were evaluated and treated according to a standard institute protocol. RESULTS: Twenty nine (45%) patients were diagnosed with ARVC/D according to TF criteria (TF+) and 35 (55%) with RV scar of undetermined origin (TF-) at the end of follow-up (64 +/ 42 months). Patients were treated with antiarrhythmic drugs, radiofrequency catheter ablation, and/or implantable cardioverter-defibrillator (ICD) implantation. VT recurrence-free survival for TF+ and TF- was 76% versus 74% at 1 year and 45% versus 50% at 4 years (P = ns). Patients with fast index VT (cycle length [CL]< or = 250 ms, n = 31) were more likely to experience a fast VT during follow-up than patients with a slow index VT (CL > 250 ms, n = 33) (61% vs 3%, P < 0.001). CONCLUSIONS: Scar-related RV VTs have a high recurrence rate in TF+ and TF- patients. Patients presenting with a fast index VT are at high risk for fast VT recurrence and may benefit most from ICD therapy. PMID- 19563367 TI - Localization of left atrial ganglionated plexi in patients with atrial fibrillation. AB - BACKGROUND: The intrinsic cardiac autonomic nervous system (ICANS), which forms a neural network, has been shown to be a critical element responsible for the initiation and maintenance of atrial fibrillation (AF). We developed a technique to localize and ablate the ganglionated plexi (GP), which serves as the "integration centers" of the ICANS. METHOD: The four major atrial GP are localized by delivering high frequency stimulation (HFS; 20 Hz, 10-150 V, 1-10 ms pulse width) to atrial tissue where GP are presumed to be located. Sites showing a parasympathetic response, which is arbitrarily defined as > or =50% increase in mean R-R interval during AF, was assigned as a GP site. Radiofrequency current is then applied to that site to eliminate the parasympathetic response. All patients received ablation of the four major atrial GP, followed by pulmonary vein antrum ablation. RESULTS: Our preliminary results showed that all the four major atrial GP can be identified in the vast majority of patients. The parasympathetic response can be eliminated by applying radiofrequency current. In the first 83 patients, the percent of patients free of symptomatic AF or atrial tachycardia after a single ablation procedure was 80% at 12 months and 86% at a mean follow up of 22 months. CONCLUSION: These results indicate additional benefits of GP ablation to PV antrum ablation and improvement with time, particularly > or = 12 months after ablation. We postulate that this late benefit may result from destruction of the autonomic neurons in the GP that cannot regenerate. PMID- 19563368 TI - Long-term follow-up on intracorneal ring segment inserted for the correction of myopia. PMID- 19563369 TI - Psychometric properties of visual functioning index using Rasch analysis. AB - PURPOSE: The visual functioning index (VFI) was one of the first questionnaires developed using classical test theory to assess outcomes of cataract surgery. However, it was not Rasch-validated. The objective of this study was to examine the psychometric properties of the VFI using Rasch analysis in patients with cataract. METHODS: The 11-item VFI was self-administered to 243 patients (mean age 73.9 years) drawn from a cataract surgery waiting list. We examined the response category thresholds, item fit statistics, differential item functioning and unidimensionality for the VFI and its three subscales. RESULTS: Category thresholds were ordered. The person separation and reliability were low, indicating the poor discriminatory ability of the VFI. No items misfit but there was suboptimal targeting of item difficulty to patient ability. On the whole the items in the VFI were too easy for the sample. Only one item showed moderate differential item functioning. CONCLUSION: The VFI does not meet the stringent requirements of the Rasch model. However adding more items to suit the more able patients with cataract as well as those awaiting second-eye cataract surgery could optimize the VFI. PMID- 19563370 TI - Ultrasound assessment of short-term ocular vascular effects of intravitreal injection of bevacizumab (Avastin((r)) ) in neovascular age-related macular degeneration. AB - PURPOSE: Angiogenic inhibitors, alone or combined with other therapies, are believed to represent a promising treatment for neovascularization in age-related macular degeneration (wet AMD). They can maintain or improve visual acuity (VA), at least for the first 2years. However, evolution to retinal atrophy cannot be ruled out and it may be useful to assess the effects of antiangiogenic therapy on retinal and choroidal circulation. METHODS: We carried out a pilot study in 15 patients with wet AMD. Time-averaged mean blood flow velocities (BFVs) in the central retinal, temporal posterior ciliary and ophthalmic arteries (CRA, TPCA and OA) were measured by ultrasound imaging before and 4weeks after a single intravitreal injection of 1.25mg bevacizumab in 0.05ml. Patients underwent two ophthalmic examinations, before and 4weeks after injection, including VA measurement and optical coherence tomography (OCT3) examination. RESULTS: In treated eyes, bevacizumab injection was followed by a significant improvement in VA (from 20/125 to 20/80; p=0.0214), and a decrease in mean central macular thickness (from 392+/-96MUm to 271+/-50MUm; p=0.0038). Mean BFV decreased by 10% in the CRA (p=0.0226), 20% in the TPCA (p=0.0026) and 20% in the OA (p=0.0003). No effect was observed in fellow eyes. CONCLUSIONS: Intravitreal bevacizumab acutely improved VA and reduced central macular thickness in wet AMD. Ultrasound imaging revealed that BFVs decreased in all retrobulbar arteries, suggesting that after local diffusion, bevacizumab exerts a short-term regional effect. Bevacizumab might therefore induce hypoperfusion of the whole eye, which may correspond to a vascular side-effect. PMID- 19563371 TI - Longterm follow-up of penetrating keratoplasty for keratoconus. AB - PURPOSE: This study aims to describe the current visual and refractive status of patients who underwent penetrating keratoplasty (PK) for keratoconus > 20 years ago and to report on the current status of their grafts. METHODS: A total of 138 eyes in 103 patients were grafted for keratoconus between August 1968 and December 1985. Patients who had not undergone retransplantation were invited to attend a clinical examination. Forty-eight patients (with 61 grafts) accepted the invitation and were examined. RESULTS: The average length of time since PK was 26.9 years (standard deviation [SD] 4.2 years, range 20.8-38.0 years, n = 61). The average graft age at examination was 82.1 years (SD 19.9 years, range 41-115 years). A total of 80% (49 of 61 grafts) of the examined eyes had a clear graft and 46% (28 of 61 eyes) had best spectacle-corrected visual acuity (BSCVA) > or = 0.5. The mean endothelial cell density per mm(2) was 894 (SD 4.6, range 470 1775). The mean central corneal thickness of the clear grafts was 0.565 mm (SD 0.048 mm). CONCLUSIONS: Penetrating keratoplasty for keratoconus has a good longterm prognosis; half of the eyes examined in this study had BSCVA > or = 0.5 at > 20 years after surgery. PMID- 19563372 TI - Increase of vascular endothelial growth factor and interleukin-6 in the aqueous humour of patients with macular oedema and central retinal vein occlusion. AB - PURPOSE: This study aimed to investigate whether vascular endothelial growth factor (VEGF) or interleukin-6 (IL-6) influence macular oedema in patients with central retinal vein occlusion (CRVO). METHODS: Sixteen consecutive patients with unilateral CRVO and macular oedema were studied, along with eight age- and sex matched patients without ischaemic ocular disease. Retinal ischaemia was evaluated from capillary non-perfusion on fluorescein angiography. Macular oedema was examined by optical coherence tomography. Aqueous humour (AH) samples were obtained during combined pars plana vitrectomy and cataract surgery, and were examined by enzyme-linked immunosorbent assay. RESULTS: Aqueous levels of VEGF and IL-6 were significantly elevated in patients compared with controls (p=0.0142 and p<0.0001, respectively). Aqueous levels of both molecules were significantly higher in patients with ischaemia than in those without ischaemia (p=0.0026 and p=0.0487, respectively). Furthermore, AH levels of VEGF and IL-6 were correlated with the severity of macular oedema (rho=0.7265, p=0.0049, rho=0.5324, and p=0.0392, respectively). CONCLUSIONS: Both VEGF and IL-6 were elevated in the AH of patients with macular oedema and ischaemic CRVO, suggesting that these molecules may be related to the increase in vascular permeability in such patients. PMID- 19563373 TI - Ultrasound biomicroscopy in two cases of ocular siderosis with secondary glaucoma. PMID- 19563374 TI - Clinical features and follow-up results of pulsating polypoidal choroidal vasculopathy treated with photodynamic therapy. AB - PURPOSE: To report on the clinical course of pulsating polypoidal choroidal vasculopathy (PCV) treated with photodynamic therapy (PDT). METHODS: A total of 63 eyes of 58 consecutive patients diagnosed with PCV, treated with PDT and followed up for at least 6months were enrolled. Best-corrected visual acuity (BCVA), fluorescein angiography and high-speed indocyanine green angiography (ICGA) using confocal scanning laser ophthalmoscopy (HRA) were performed. RESULTS: Of the 63 PCV eyes, 14 eyes (22.2%) of 14 patients were classified as having pulsating PCV. The mean age of pulsating PCV patients was 60.6+/-7.0years (48-69years), which was younger than non-pulsating PCV patients (65.7years, p=0.035). The mean follow-up period was 23.9+/-10.7months, and PDT was administered 1.6+/-0.9 times to pulsating PCV patients. The mean logMAR BCVAs were 0.85+/-0.47 at presentation and 0.71+/-0.52 at final examination. Extensive haemorrhagic events were more common in pulsating than in non-pulsating PCV patients (57.1% versus 26.5%, p=0.032). However, the risk of haemorrhage within 3months of PDT was similar for both pulsating PCV and the remaining patients (14.3% versus 20%, p=0.723). CONCLUSION: Pulsating PCV showed distinctive features including a relatively younger patient age at presentation, and a haemorrhagic tendency (especially extensive). However, the use of PDT did not directly increase the risk of haemorrhage in pulsating PCV patients. PMID- 19563376 TI - Weighing the evidence for treatment of chronic kidney disease mineral and bone disorder. PMID- 19563375 TI - In vitro evaluation of bevacizumab toxicity on a retinal ganglion cell line. AB - PURPOSE: The effects of bevacizumab on cell viability and proliferation in a commonly used retinal ganglion cell line, RGC-5, were examined. METHODS: RGC-5 cells were exposed to 0.1 mg/ml, 1 mg/ml and 2 mg/ml of commercially available bevacizumab in vitro. To examine the specificity of effects, cells were also cultured with increasing and comparable concentrations of proteins (increasing the concentration of proteins in the culture media by 0.1 mg/ml, 1 mg/ml and 2 mg/ml by using additional fetal bovine serum [FBS] and bovine serum albumin [BSA]). Cell proliferation was assessed using a WST-1 kit, crystal violet staining and bromodeoxyuridine (BrdU) incorporation. Cytotoxic effects were assessed by quantifying cell numbers in proliferation-deficient RGC-5 following exposure to bevacizumab using the WST-1 kit, microscopic examination of cells stained with propidium iodide (PI) cells and flow cytometry for differential staining with PI. RESULTS: Bevacizumab was not toxic to RGC-5 cells in the tested concentrations. It had a stimulatory effect on cell proliferation. A stimulatory effect on proliferation was also noted when equivalent amounts of proteins from FBS or BSA were used, which suggests that bevacizumab may stimulate proliferation non-specifically by increasing the protein contents of the cell growth environment. CONCLUSIONS: Results suggest that intravitreal injection of bevacizumab could alter the internal milieu of the eye by increasing protein concentrations to elicit functional responses in retinotypic cells. This may be especially relevant for cells outwith the control of vascular endothelial growth factor. PMID- 19563377 TI - Review article: Early detection of chronic kidney disease in Australia: which way to go? AB - Early detection of chronic kidney disease (CKD) followed by appropriate clinical management appears the only means by which the increasing burden on the health care system and affected individuals will be reduced. The asymptomatic nature of CKD means that early detection can only occur through testing of individuals. The World Health Organization principles of screening for chronic disease can now be largely fulfilled for CKD. The risk groups to be targeted, the expected yield and the tests to be performed are reviewed. For a screening programme to be sustainable it must carry a greater benefit than risk of harm for the participant and be shown to be cost-effective from the community point of view. Whole population screening for CKD is impractical and is not cost-effective. Screening of those at increased risk of CKD could occur either through special events run in the community, workplace or in selected locations such as pharmacies or through opportunistic screening of high-risk people in general practice. Community screening programmes targeted at known diabetics, hypertensives and those over 55 years have been described to detect 93% of all CKD in the community. The yield of CKD stages 3-5 from community screening has been found to vary from 10% to 20%. The limitations of screening programmes including the cost and recruitment bias are discussed. The most sustainable and likely the most cost efficient model appears to be opportunistic general practice screening. The changing structure of general practice in Australia lends itself well to the requirements for early detection of CKD. PMID- 19563378 TI - Review article: Chronic kidney disease-mineral bone disorder: have we got the assays right? AB - Parathyroid hormone (PTH) and vitamin D [calcidiol or 25(OH)D] assays are used to diagnose chronic kidney disease-mineral bone disorder, and to guide and monitor interventions. Unfortunately, both assays have limitations. These limitations include analytical variability because of a lack of assay standardization, different calibrators used for each kit and differences in antibody specificity. For PTH assays, this leads to variability in the assessment of fragments and differences between results among kits from different manufacturers. For vitamin D, this leads to differences in the ability to detect D(2) and D(3) derivatives and differences among assays. For PTH, these problems are amplified by collection and handling differences and, for vitamin D, by diurnal and seasonal variation. Understanding these limitations should help clinicians to appropriately interpret results; attempts to develop universal standards to minimize these limitations may enhance the ability of these hormone measurements to predict underlying disease states. PMID- 19563379 TI - Serum phosphate is an important determinant of corrected serum calcium in end stage kidney disease. AB - BACKGROUND: Approximately 12% of bound blood calcium is linked to various anions including phosphate. In patients with end-stage kidney disease (ESKD), serum phosphate is highly variable. We propose that establishing a formula to calculate albumin- and phosphate-corrected total calcium would be more appropriate to estimate free calcium in ESKD patients. METHODS: In 82 haemodialysis patients, serum ionized calcium (Ca(ion)) and pH were measured by blood gas analyser with ion-selective electrodes at the point-of-care, while bicarbonate, phosphate, albumin, magnesium and total calcium (Ca(tot)) were measured at the central laboratory. Linear regression analysis of measured variables was used to best fit adjusted calcium versus Ca(ion). RESULTS: The most parsimonious multiple linear regression model (r(2) = 0.81) of variables associated with Ca(ion) included Ca(tot) (coeff 0.820, P < 0.0001), albumin (coeff -0.016, P < 0.0001) and phosphate (coeff -0.063, P < 0.002). Modelling of available variables yielded the following equation to adjust calcium for albumin and phosphate: Ca(albPh) = Ca(tot) + (0.015 x (40 - [albumin]) + 0.07 x (1.5 - [phosphate])). At an ambient albumin of 40 g/L, Ca(albPh) would be 0.07 mmol/L lower than Ca(tot) for every mmol/L of phosphate. In vitro data using three different albumin levels and increasing phosphate concentrations demonstrated this relationship, with the slope of the phosphate effect being stronger at lower albumin concentrations. CONCLUSION: Because guidelines recommendations indicate that corrected serum calcium should be maintained within the normal range in ESKD patients, inclusion of phosphate to correct Ca(tot) in these patients may have clinical implications on the choice of phosphate binders and the prescription of vitamin D or calcimimetic agents. PMID- 19563380 TI - Review article: Getting the balance right: assessing causes and extent of vascular calcification in chronic kidney disease. AB - Vascular calcification is part of the definition of chronic kidney disease mineral and bone disorder (CKD-MBD). It is also a surrogate parameter of cardiovascular and all-cause mortality risk in the CKD population. However, vascular calcification is not a homogenous entity, but a rather complex manifestation influenced by derangements of calcium and phosphate homeostasis, by dysregulated calcification inhibitors and promoters, and by the type of arterial disease (atherosclerosis vs arteriosclerosis). Despite the clear-cut risk association between the presence of vascular calcification and mortality, it is currently not well defined, how this knowledge about calcification should be translated into active clinical management. Further, the choice of the appropriate imaging test is a matter of debate. This article attempts to provide an update on insights into the pathophysiology of vascular calcification processes and a subjective view of the clinical consequences of management of CKD patients at risk. PMID- 19563381 TI - Review article: Bone density in patients with chronic kidney disease stages 4-5. AB - Skeletal fractures are common in dialysis patients. Bone density measurements predict fractures in the general population, but in patients with chronic kidney disease (CKD) there is limited ability to predict fractures with these techniques. A review of the literature in patients with CKD stage 5 shows that bone density in the cortical bone of the radius or hip is generally lower compared with normal reference ranges by about one to two standard deviations, whereas in the spine bone density tends to be closer to the average expected values. The bone density is not able to predict the underlying histology in patients with renal osteodystrophy. The serum PTH levels are inversely related to bone density in some studies, and not related in other studies, but none found a positive association of serum parathyroid hormone (PTH) and bone density. There are very few data about longitudinal changes of bone density in these patients, and also inadequate studies of whether osteoporosis medications are effective in this population. Therefore, at this time the clinical utility of bone density measurements in patients with CKD-5 is uncertain. PMID- 19563382 TI - Review article: Bone biopsy in chronic kidney disease: patient level end-point or just another test? AB - The reduction of renal function in chronic kidney disease leads to disturbed calcium and phosphorus metabolism, impaired action of calcitriol, increased parathyroid hormone, FGF-23 levels and ultimately bone disorders. These disturbances have been traditionally termed renal osteodystrophy, which evaluation and diagnosis require a bone biopsy. In the last four decades, researchers from different countries have developed new techniques and have introduced concepts that allowed the development of bone histomorphometry, considered the key tool to study bone metabolism, remodelling and structure. In this review we focus on the relevance of bone biopsy and its respective histomorphometric analysis to help nephrologists to evaluate patients with chronic kidney disease. PMID- 19563384 TI - Review article: Addressing risk factors in chronic kidney disease mineral and bone disorder: can we influence patient-level outcomes? AB - Chronic kidney disease mineral and bone disorder (CKD-MBD), characterized by disturbances of calcium/phosphate/parathyroid hormone, bone abnormalities and vascular and soft tissue calcification, is highly prevalent in CKD and is a strong, independent predictor of bone fracture, cardiovascular disease and death. Clinical practice guidelines, such as the Kidney Disease Outcomes Quality Initiative (KDOQI) and Caring for Australasians with Renal Insufficiency (CARI), support the use of phosphate binders, vitamin D compounds and calcimimetics for treatment of CKD-MBD and recommend stringent targets for serum calcium, phosphate and parathyroid hormone. However, these recommendations are based primarily on the results of observational cohort studies and randomized controlled trials employing surrogate outcome measures. The aim of this paper is to review the available evidence addressing whether therapeutic strategies targeting CKD-MBD and its surrogate outcome measures appreciably influence patient-level outcomes ('hard' clinical end-points). PMID- 19563385 TI - Review article: Risks of coronary artery calcification in chronic kidney disease: do the same rules apply? AB - Atherosclerosis, once present, in the intimal and medial spaces of the blood vessel wall becomes calcified due to a variety of cellular and metabolic processes. Patients with chronic kidney disease (CKD) appear to have both accelerated and amplified vascular calcification compared with the general population. Calcium deposition within vascular tissue in the form of calcium hydroxyapatite crystals appears to be a permanent step in the mature atherosclerotic plaque, and to date has not been found to be reversible or modifiable with common treatments for atherosclerosis or dialysis management strategies. Densely calcified lesions with a circumferential arc of calcium around the vessel wall may be severely stenotic, however, are unlikely to develop symptomatic plaque rupture with an acute coronary syndrome. For that reason, the expected outcomes of atherosclerotic therapies in patients with CKD bone and mineral disorder have not followed the same rules of evidence. This paper will review the differences between the CKD and general population with respect to vascular calcification and the observed natural history in observational and interventional studies. PMID- 19563386 TI - Review article: Managing bone complications after kidney transplantation. AB - Chronic kidney disease mineral and bone disorder (CKD-MBD) describes the laboratory, bone and vascular abnormalities that exist in patients with CKD stages 3-5D and that may persist after transplantation. Persisting abnormalities of bone turnover and abnormal mineralization, together with bone mineral density (BMD) loss from glucocorticoids, may all predispose to a loss of structural integrity and increased fracture risk in kidney and kidney pancreas recipients. Vitamin D, calcitriol, calcitonin and bisphosphonates have all been used to preserve BMD following transplantation, despite a lack of safety data and the potential for some of these drugs to cause harm. A limited number of post transplant studies utilizing these drugs have not yet documented improved fracture prevention or fracture-related mortality and have not considered allocation based on risk factors for fracture or markers of bone turnover. Targeted allocation of the available therapies based on a stratification of risk appears warranted. This might be achieved using an algorithm incorporating BMD, X ray evaluation, laboratory investigations including bone turnover markers and the assessment of standard fracture risk factors at the time of and soon after transplantation. This approach, which is similar to protocols used in the general population, may result in more effective management of patients and fewer adverse effects such as adynamic bone disease. Although BMD is a surrogate for fracture risk in the general population it is not validated in this transplant population. Consequently, such an approach should be confirmed by studies that include bone biopsy data and an evaluation of patient level outcomes. PMID- 19563383 TI - Review article: Biomarkers of clinical outcomes in advanced chronic kidney disease. AB - Chronic kidney disease (CKD) is a complex condition, where the decrease in kidney function is accompanied by numerous metabolic changes affecting virtually all the organ systems of the human body. Many of the biomarkers characteristic of the individually affected organ systems have been associated with adverse outcomes including higher mortality in advanced CKD, whereas in persons without CKD these biomarkers may have no bearing on survival. It is believed that the high mortality seen in CKD is a result of several abnormalities conspiring to induce or aggravate a heightened degree of cardiovascular morbidity and predisposition to wasting syndrome. Not all the biomarkers may, however, be causally responsible for the adverse outcomes associated with them. We review various biomarkers of protein-energy wasting, inflammation, oxidative stress, potassium disarrays, acid base disorders, bone and mineral disorders, glycemic status, and anemia. Although all of these biomarkers have shown associations with worsened outcomes in CKD, markers of protein-energy wasting, especially serum albumin, remain the strongest predictor of survival in CKD patients, especially those undergoing maintenance dialysis treatment. We also review the putative pathophysiologic mechanisms behind these associations, and present potential therapeutic interventions that could result in remedies to improve poor clinical outcomes in CKD, pending the results of current and future controlled trials. PMID- 19563387 TI - Review article: Patient-level outcomes: the missing link. AB - Treatment of chronic kidney disease (CKD) may be life-saving, but can disrupt every aspect of a patient's life and the lives of family members. Many patients with CKD are elderly with significant comorbidities and sometimes therapies to improve survival may be less important than those that improve or maintain quality of life. In this setting, patient-level benefits become particularly important goals of therapy. Randomized controlled trials (RCT) are also essential to justify expensive therapies, such as medications used in the treatment of CKD mineral and bone disorders. Surprisingly, data to support the efficacy of these drugs for patient-level outcomes remains limited. In fact, fewer RCT are conducted in renal medicine than in any other medical specialty and reliance is often placed on association data and the assessment of intermediate and biochemical end-points. While some of these may prove to be valid surrogates for clinically important outcomes, some may not. Inclusion of patient-level outcomes in clinical research provides a missing link that can inform a more comprehensive approach to clinical practice and patient care. Incorporating measures of health related quality of life into clinical trials can make outcomes more relevant and may be relatively simple. This paper provides examples of reliable, validated instruments to measure health-related quality of life domains and functional status, together with practical instructions for their use. Most could be incorporated into RCT of CKD mineral and bone disorder treatments. Inclusion of outcomes that are perceived by patients to be significant should become standard practice in renal medicine and in clinical renal research. PMID- 19563388 TI - Recurrent sarcoidosis with psoas muscle granuloma and hypercalcaemia in a patient on chronic haemodialysis. PMID- 19563390 TI - Uric acid and risk of myocardial infarction, stroke and congestive heart failure in 417,734 men and women in the Apolipoprotein MOrtality RISk study (AMORIS). AB - OBJECTIVES: Few studies have simultaneously analysed the influence of elevated serum uric acid (UA) on acute myocardial infarction (AMI), ischaemic and haemorrhagic stroke (IS, HS) and congestive heart failure (CHF) in large healthy populations. We, here, examine UA as a risk factor for AMI, stroke and CHF by age and gender in the Apolipoprotein MOrtality RISk (AMORIS) Study. DESIGN: Prospective study (11.8 years, range 7-17) of fatal and nonfatal acute myocardial infarction, stroke and CHF through linkage with Swedish hospital discharge and mortality registers. SETTINGS: Measurements of uric acid in 417,734 men and women from health check-ups in Stockholm area. RESULTS: There was a gradual increase in risk of AMI, stroke and CHF by increasing UA levels. Women had a stronger relationship between UA and both AMI and IS than men. Predictions of AMI were at least as powerful in the elderly as in the young, but not so for IS. Associations were markedly attenuated when adjusted for total cholesterol, triglycerides, hospital hypertension and diabetes status. The association between UA and HS was U-shaped in both genders. CHF was more strongly related to UA than AMI and stroke and less affected by the adjustment factors. CONCLUSIONS: Already moderate levels of UA appear to be associated with an increased incidence of AMI, stroke and CHF in middle-aged subjects without prior cardiovascular disease. These associations seem to increase gradually from lower to higher levels of UA. UA may be an important complementary indicator of cardiovascular risk in the general population. PMID- 19563389 TI - Telomere length and progression of diabetic nephropathy in patients with type 1 diabetes. AB - OBJECTIVES: To determine whether short telomere length of blood leucocytes from patients with type 1 diabetes is associated with or predictive of progression of diabetic nephropathy. DESIGN AND METHODS: Two consecutive DNA samples were obtained from 132 patients from the nationwide Finnish Diabetic Nephropathy Study with type 1 diabetes. Control DNA samples were taken from 44 healthy blood donors. Telomere length was measured by Southern blot. Patients were divided into three groups according to their urinary albumin excretion rate (AER): 48 patients with normoalbuminuria (AER < 20 microg min(-1)); seven patients with microalbuminuria (AER > or = 20 microg min(-1) <200 microg min(-1)) and 77 patients with macroalbuminuria (AER > or = 200 microg min(-1)). Progression was defined as a change in albuminuria to a higher level. RESULTS: Progression occurred in 21 patients. Progressors had shorter mean telomere length (8.1 +/- 0.7 kb, mean +/- SD; P = 0.017) and higher percentage of short telomeres (32.0 +/ 8%, P = 0.002) than nonprogressors (8.5 +/- 0.7 kb and 27 +/- 7.2%, respectively). Thus, both shorter telomeres (HR = 0.190, 95%CI 0.065-0.558, P = 0.0025) and higher proportion of short telomeres (HR = 1.115, 1.039-1.195, P =0.0023) were independent predictors of diabetic nephropathy. Telomere length was not associated with the degree of albuminuria and was not different in patients with type 1 diabetes compared with healthy controls. CONCLUSIONS: Short telomeres are independent predictors of progression of diabetic nephropathy in patients with type 1 diabetes. PMID- 19563391 TI - Fitness and abdominal obesity are independently associated with cardiovascular risk. AB - OBJECTIVES: To examine the relationship between cardiovascular fitness (VO(2)max) and abdominal obesity (waist circumference) and individual cardiovascular disease (CVD) risk factors, as well as a clustered risk factor profile, and to study the impact of gender, age and smoking on these relationships. DESIGN: Cross sectional. SETTING: Astrand Laboratory of Work Physiology, Swedish School of Sport and Health Sciences, Stockholm, Sweden. SUBJECTS: Men (n = 781) and women (n = 890) from two random population-based samples of Swedish women and men aged 20 to 65 years. MAIN OUTCOMES: Odds ratios. RESULTS: Each unit of higher fitness was associated with a decrease in all individual risk factors ranging from 2% to 4% independent of waist circumference, each unit of higher waist circumference was associated with an increased risk ranging from 2% to 5% independent of fitness. For clustering of three or more of the risk factors, each unit of fitness was associated with a 5% decrease in risk and each unit of waist circumference with a 5% increase in risk. The clustered risk was higher in unfit participants who were older or smoked daily, regardless of waist circumference. Obese participants were at higher risk if they were men or older, regardless of fitness level. However, neither a higher fitness level nor lean status reduced the risk associated with smoking. CONCLUSIONS: Higher fitness and lower waist circumference are each independently associated to a similar extent with a lower CVD risk. Simultaneous evaluation of both fitness and abdominal obesity status in clinical practice is important. PMID- 19563392 TI - ANTI-HSP60 and ANTI-HSP70 antibody levels and micro/ macrovascular complications in type 1 diabetes: the EURODIAB Study. AB - OBJECTIVES: The heat shock proteins 60 and 70 (HSP60, HSP70) play an important role in cytoprotection. Under stress conditions they are released into the circulation and elicit an immune response. Anti-HSP60 and anti-HSP70 antibody levels have been associated with cardiovascular disease. Type 1 diabetes is associated with a greatly increased risk of micro- and macrovascular complications. Therefore, we investigated whether anti-HSP60 and anti-HSP70 antibody levels were associated with micro- and macrovascular complications in type 1 diabetic patients. DESIGN: A cross-sectional nested case-control study from the EURODIAB Study of 531 type 1 diabetic patients was performed. SUBJECTS: Cases (n = 363) were defined as those with one or more complications of diabetes; control subjects (n = 168) were all those with no evidence of any complication. We measured anti-HSP60 and anti-HSP70 antibody levels and investigated their cross-sectional associations with diabetic complications. RESULTS: Anti-HSP70 antibody levels were significantly greater in control than in case subjects, whereas anti-HSP60 antibody levels were similar in the two groups. In logistic regression analysis, anti-HSP70 levels in the upper quartiles were associated with a 47% reduced odds ratio of micro/macrovascular complications, independently of conventional risk factors, markers of inflammation and endothelial dysfunction [odds ratio (OR) = 0.53, 95% confidence intervals (CI): 0.28-1.02]. CONCLUSIONS: In this large cohort of type 1 diabetic subjects, we found an independent and inverse association between serum anti-HSP70 antibody levels and diabetic micro/macrovascular complications. This suggests that anti-HSP70 antibody levels may be a novel marker of protection from chronic diabetic complications. PMID- 19563393 TI - Lipid lowering versus pleiotropic effects of statins on skin microvascular function in patients with dysglycaemia and coronary artery disease. AB - OBJECTIVES: To investigate the impact of lipid lowering therapy by different means on skin microvascular function in patients with dysglycaemia and coronary artery disease (CAD). DESIGN AND SETTING: Thirty-six patients were randomized to simvastatin 80 mg daily (S80, n = 19) or ezetimibe 10 mg and simvastatin 10 mg daily (E10/S10, n = 17) for 6 weeks. Skin microvascular function was assessed by laser Doppler fluxmetry (LDF) at rest, following arterial occlusion (peak postocclusive LDF) and following local heating on the forearm (heat arm LDF) and foot (heat foot LDF). LDF parameters and serum lipids were evaluated at baseline and follow-up. RESULTS: At follow-up, LDL cholesterol had decreased from 3.1 (2.7 3.5) to 1.6 (1.5-1.8) (mmol L(-1)) and 3.0 (2.4-3.9) to 1.3 (1.1-1.8) (mmol L( 1)) in the E10/S10 and S80 groups respectively. In the entire study group (n = 32), LDF parameters increased significantly; postocclusive LDF from 22 (17-27) to 26 (21-32) perfusion units (PU) (P < 0.001), heat foot LDF from 61 (44-82) to 66 (45-83) PU (P < 0.001) and heat arm LDF from 60 (48-121) to 75 (54-125) PU (P < 0.01). The changes in LDF parameters did not differ between the E10/S10 and S80 groups. CONCLUSIONS: Lipid lowering improves microvascular function in patients with dysglycaemia and CAD. The data suggest that lipid lowering per se is more important than pleiotropic effects of statins for this effect. PMID- 19563394 TI - Classification of OprD sequence and correlation with antimicrobial activity of carbapenem agents in Pseudomonas aeruginosa clinical isolates collected in Japan. AB - A total of 99 clinical isolates of metallo-ss-lactamase-negative Pseudomonas aeruginosa collected in Japan between 1998 and 2001 were studied for their susceptibilities to carbapenem agents and corresponding oprD gene mutations. The OprD sequence of each strain was grouped into two major classes, based on the pattern of alterations. Eighty strains (80.8%) were so-called 'full length type', whose OprD proteins were fully encoded. The remaining 19 strains (19.2%) were so called 'defective type', which possessed deletions or major alterations that might cause conformational changes in the OprD porin protein. The changes in 'defective type' strains led to 15-, 17- and 23-fold increases in the geometric mean MIC for imipenem, meropenem and biapenem compared with 'full length type' strains, respectively. 'Full length type' strains were further classified into six carbapenem susceptible types with the exception of four carbapenem-resistant subtypes with additional amino acid substitutions at D43, G183, R154, G314, G316. However, 'defective type' strains were classified into four types as follows: 10 strains which contained a stop codon within the coding region; six strains which contained IS; one strain with a short deletion near the C-terminal domain; and two strains without a stop codon in the sequenced region. Western blot analysis using OprD antibody showed that binding abilities of OprD proteins against 'full length type' strains were normal, whereas those against 'defective type' strains were lost without exception. These results indicate that OprD structure and antimicrobial activities for carbapenem agents proved to be highly correlated in P. aeruginosa. PMID- 19563395 TI - Development of recombinant OmpA and OmpB proteins as diagnostic antigens for rickettsial disease. AB - In this study the diagnostic potential of Rickettsia conorii recombinant antigens was analyzed. For this, site-specific PCR primers were used to clone the OmpA and OmpB genes of R. conorii into pMAL-c2X plasmids. Six fragments of OmpA and four of OmpB were expressed as fusion proteins with maltose-binding protein in Escherichia coli. OmpA(1350-1784), OmpB(801-1269,) and OmpB(1227-1634) regions from truncated proteins were selected as diagnostic candidate antigens by ELISA using control sera. ELISA results of three antigens were compared to the results obtained by using a commercial ELISA kit which contained whole OmpA and OmpB antigens from R. conorii. For this analysis, 40 serum samples taken from febrile patients and uninfected controls were tested. Of the 20 R. conorii test results which were positive with the commercial kit, 18 were shown to be positive by ELISA using OmpA(1350-1784) (a sensitivity of 90%). The specificity of the ELISA was 100%; all of the 20 samples shown to be negative using the commercial kit were also negative in our assay. The sensitivities of the ELISA using the OmpB(801-1269) and OmpB(1227-1634) were 90% and 95%, respectively. The specificities of the OmpB(801-1269) and the OmpB(1227-1634) were 100% and 95%, respectively. These results suggest that specific regions of OmpA and OmpB effectively detect antibodies against R. conorii, and the truncated recombinant antigens could be used for development of diagnostic tools for rickettsial disease. PMID- 19563396 TI - Rapid, sensitive and simple detection method for koi herpesvirus using loop mediated isothermal amplification. AB - New methods were developed for the detection of koi herpesvirus (KHV, CyHV-3) by LAMP, which were compared with the PCR for specificity and sensitivity. We designed two primer sets targeting a specific sequence within the 9/5 PCR amplicon (9/5 LAMP) and the upper region of the SphI-5 PCR amplicon (SphI-5 LAMP), including a sequence highly conserved among the strains. The amplification was monitored in real-time based on the increase in turbidity, with magnesium pyrophosphate as the by-product. The reactions were carried out under isothermal conditions at 65 degrees C for 60 min. The detection limit of both LAMP was six copies, equal to the modified SphI-5 PCR. No cross-reactivity with other fish pathogenic viruses and bacteria was observed. SphI-5 LAMP was found to have a quicker response in terms of the reaction velocity than 9/5 LAMP. Therefore, we consider SphI-5 LAMP to be superior for routine use. Additionally, LAMP was found applicable to crude extract from gills and other organs. LAMP methods are superior in terms of sensitivity, specificity, rapidity and simplicity, and are potentially a valuable diagnostic tool for KHV infections. PMID- 19563397 TI - Improved cellular immune response elicited by a ubiquitin-fused ESAT-6 DNA vaccine against Mycobacterium tuberculosis. AB - The present study evaluated the immune response elicited by a ubiquitin-fused ESAT-6 DNA vaccine against Mycobacterium tuberculosis. BALB/c mice were vaccinated with plasmid DNA encoding ESAT-6 protein, ubiquitin-fused ESAT-6 DNA vaccine (UbGR-ESAT-6), pcDNA3-ubiquitin and blank vector, respectively. ESAT-6 DNA vaccine immunization induced a Thl-polarized immune response. The production of Thl-type cytokine (IFN-gamma) and proliferative T-cell responses was enhanced significantly in mice immunized with UbGR-ESAT-6 fusion DNA vaccine, compared to non-fusion DNA vaccine. This fusion DNA vaccine also resulted in an increased relative ratio of IgG(2a) to IgG(l) and the cytotoxicity of T cells. Thus, the present study demonstrated that the UbGR-ESAT-6 fusion DNA vaccine inoculation improved antigen-specific cellular immune responses, which is helpful for protection against tuberculosis infection. PMID- 19563398 TI - Effect of GM-CSF on cytokine induction by soluble beta-glucan SCG in vitro in beta-glucan-treated mice. AB - SCG is a 6-branched 1,3-beta-D-glucan, which are major cell wall structural components in fungi. Leukocytes from DBA/1 and DBA/2 mice are highly sensitive to SCG, producing cytokines such as GM-CSF, IFN-gamma, TNF-alpha and IL-12p70, but not IL-6. GM-CSF plays a key biological role in this activity. In the present study, we examined the effect of giving i.p. SCG to DBA/2 mice on cytokine production in vitro. SCG was given i.p. to DBA/2 mice on day 0. Splenocytes were prepared on day 7 and cultured in the presence of SCG in vitro. The levels of cytokine production induced by SCG in vitro were lower in the cells from SCG treated mice than in control mice. Expression of the beta-glucan receptor, dectin 1, in SCG-treated mice was comparable with that shown in control mice. However, the consumption of exogenously added rmGM-CSF in vitro was observed in SCG treated mice. The addition of a large amount of rmGM-CSF to the culture medium resulted in larger amounts of TNF-alpha and IL-6 in SCG-treated mice than in normal mice. These results suggested that GM-CSF was closely related with the reactivity of beta-glucan. Giving SCG increased the number of macrophages and granulocytes in the spleen. These results suggested that in SCG-treated mice, a change of cell population would be related to modulation of the profile of cytokine production induced by SCG in vitro. PMID- 19563399 TI - Human plasmacytoid dendritic cells express an atrial natriuretic peptide receptor, guanylyl cyclase-A. AB - Atrial natriuretic peptide is a cardiovascular hormone secreted mainly by the cardiac atria and regulates the volume-pressure homeostasis. The action of ANP is mediated by GC-A. We previously reported that human monocyte-derived dendritic cells express GC-A and respond to ANP with polarization toward a Th2-inducing phenotype. In the present study, we explored the possibility that pDC are subjected to immunoregulation via the ANP/GC-A system. We examined GC-A expression on blood pDC and found that GC-A was not expressed on fresh pDC but was induced after stimulation with CpG-oligodeoxynucleotide AAC-30, IL-3, or interleukin-3 plus CD40 ligand. Activated pDC responded to ANP with an increase in cGMP production, indicating that GC-A expressed on pDC was functional. We investigated whether tonsillar pDC express GC-A by immunohistochemistry and immunofluorescence staining. We found that GC-A(+) HLA-DR(+) cells were present in the T-cell areas and the perivascular areas. Flow cytometric analysis with tonsillar cells confirmed that lineage(-) CD123(high) pDC express GC-A. These results indicate that the ANP/GC-A system is involved in immune regulation through pDC in secondary lymphoid organs. PMID- 19563401 TI - Distinct genotype and antigenicity among genogroup II sapoviruses. AB - SaV, a pathogen of acute gastroenteritis, is divided into five genogroups, GI to GV. However, the relation between SaV antigenicity and genetic clusters is not fully understood. We have recently identified two GII SaV strains, Mc10 and C12, which are grouped into the same cluster based on the polymerase but are grouped into distinct clusters based on the capsid. To evaluate the difference in antigenicity between these two strains, VLP were expressed in mammalian cells. An antigen ELISA demonstrated for the first time that strains in the same GII SaV genogroup, but within different clusters, have distinct antigenicities. PMID- 19563400 TI - Association between antibody response against cytomegalovirus strain-specific glycoprotein H epitopes and HLA-DR. AB - The gH of CMV is a major target for strain-specific neutralizing antibodies. To verify whether there is a correlation between HLA-DR type and strain-specific antibodies, antibodies against CMV gH in potential donors and recipients for renal transplantation were investigated. Among 471 subjects, 404 (86%) showed reactivity to CMV gH, but no antibodies against gH were detected in 67 (14%) subjects. The positive rates were over 80% in most HLA subpopulations. Fewer subjects with HLA-DR10 and DR11 had antibodies to CMV gH than did those without HLA-DR10 and DR11. HLA-DR10 and DR11 may be associated with fewer/non-responders for strain-specific neutralizing antibodies. PMID- 19563403 TI - Death comes early: apoptosis observed in ENS precursors. AB - Cell death is a physiological and fundamental process in normal organogenesis. During the development of the nervous system, cell death or apoptosis occurs in early and late developmental time periods, affecting neural precursors and neurons respectively. In the development of the enteric nervous system (ENS), however, apoptosis of neurons has not been detected, a feature unique to enteric neurons. In this issue of Neurogastroenterology and Motility, Wallace et al. focused on an early phase of ENS development and identified apoptotic cell death in vagal neural crest cells, the primary cellular source for the ENS. Introduction of an antiapoptotic molecule in the vagal neural crest and its derivatives resulted in the overproduction of neurons in the foregut. Thus, unlike the neurons themselves, ENS precursors do undergo apoptosis, which may, by regulating the size of the ENS precursor pool, be a crucial factor in determining the final cell number in the ENS. PMID- 19563405 TI - Comparative clinicopathological study of primary CNS diffuse large B-cell lymphoma and intravascular large B-cell lymphoma. AB - Primary CNS diffuse large B-cell lymphoma (CNS DLBCL) is confined to the CNS, and constitutes a distinct entity. In the present study a series of 40 Japanese patients with CNS DLBCL who presented with neurological, but not systemic symptoms, was reviewed. Median survival was 18.7 months. CD5, CD10, Bcl-6, MUM-1, and Bcl-2 were positive in 30%, 10%, 84%, 100%, and 93% of patients, respectively. All CD10-negative patients had non-germinal center B-cell type. There was no significant difference in survival among the immunophenotypic subgroups. CNS DLBCL appeared to be homogenous as a group, which prompted the comparison with another distinct extranodal entity, intravascular large B-cell lymphoma (IVLBCL) in Japanese patients. CNS DLBCL patients did not differ in age, sex, or immunophenotype, including CD5 positivity, from IVLBCL patients, but were significantly less likely to have poor prognostic parameters than IVLBCL patients: the international prognostic index score was low or low-intermediate in 86% of CNS DLBCL patients and high or high-intermediate in 98% of IVLBCL patients. Notably, despite this difference, their survival curves almost overlapped. The present study highlights the issue of clinical distinctiveness of aggressive extranodal lymphomas, the peculiar migration and localization of which should be further clarified. PMID- 19563406 TI - Hepatocellular apoptosis associated with cytotoxic T/natural killer-cell infiltration in chronic active EBV infection. AB - The aim of the present study was to identify the mechanism of hepatocellular apoptosis induced by EBV-infected cytotoxic T/natural killer (NK) cells in chronic active EBV infection (CAEBV). Eight patients with CAEBV were studied, and infected T-cell expansion and NK-cell expansion were detected in four patients each. Biopsy or necropsy was performed on lymph node, liver, or spleen, and each specimen was subjected to immunohistochemical double staining of CD3 plus caspase 3 with the addition of cytotoxic markers of T-cell restricted intracellular antigen-1 (TIA-1), perforin, and granzyme B, as well as EBV in situ hybridization (EBV-ISH). In the liver, some of the infiltrating CD3-positive lymphocytes stained positively for EBV-ISH and cytotoxic markers. Double staining of CD3 plus caspase-3 indicated caspase-3 positive hepatocytes with apoptotic features, accompanied by extensive infiltration of CD3-positive cells, which were directly attached to the apoptotic caspase-3 positive hepatocytes. In contrast, far fewer cells stained positive for caspase-3 in lymph node and spleen than in liver. The present findings suggest that in patients with CAEBV, cytotoxic T/NK cells may directly induce hepatocytes to undergo apoptosis more frequently than they do cells in other organs of the reticulo-endothelial system. PMID- 19563404 TI - Japanese herbal medicine in functional gastrointestinal disorders. AB - Management of functional gastrointestinal disorders is hindered by both poor efficacy and adverse effects of traditional pharmacological therapy. Herbal medicine may be an attractive alternative based on the perception of its 'natural' approach and low risk of side effects; however, the lack of standardization of drug components has limited the ability to perform rigorous clinical studies in Western countries. Japanese herbal medicine (JHM) is a standardized form of herbal medicine with regards to the quality and quantities of ingredients. While extensively studied and widely used in Asia, there is a paucity of data upon which physicians in other parts of the world may draw conclusions regarding the effectiveness of herbal medicine for gastrointestinal disorders. The aim of this study was to summarize the most recent developments in JHM for treatment of functional gastrointestinal disorders. Animal and human studies were systematically reviewed to identify published data of JHM used for treatment of gastrointestinal disorders. The herbal components of JHM were examined. Results describing the physiological and clinical effects of JHM were abstracted, with an emphasis on functional gastrointestinal disorders. JHM are associated with a variety of beneficial physiological on the gastrointestinal system. Patient-based clinical outcomes are improved in several conditions. Rikkunnshi-to reduces symptoms and reverses physiological abnormalities associated with functional dyspepsia, while dai-kenchu-to improves symptoms of postoperative ileus and constipation in children. This updated summary of JHM in the field of gastrointestinal disorders illustrates the potential for herbal medication to serve a valuable role in the management of patients with functional gastrointestinal disorders. PMID- 19563407 TI - Clinicopathological significance of DEK overexpression in serous ovarian tumors. AB - To investigate the significance of DEK protein expression in ovarian lesions, a total of 113 ovarian serous tumors, including 62 serous cystadenocarcinomas and 19 serous borderline tumors, were studied on immunohistochemistry. For comparison, 32 benign serous tumors, including 12 serous papillary cystadenomas, 10 serous cystadenomas, and 10 serous surface papillomas, were also included. DEK was positive in 93.5% of serous cystadenocarcinomas (58/62), 63.2% of serous borderline tumors (12/19), and weakly positive in 15.6% of benign serous tumors (5/32). The strong positive signal was detected only in serous adenocarcinomas (80.6%, 50/62) and borderline tumors (21.1%, 4/19), but no serous benign tumors were strongly positive (0%, 0/32). Meanwhile, the strong positivity rate of DEK protein was significantly higher in grade 2 and grade 3 than in grade 1 ovarian cancers (P < 0.05), but there was no significant association between DEK protein expression level and International Federation of Gynecology and Obstetrics (FIGO) stage of serous ovarian adenocarcinoma (P > 0.05). In summary, DEK plays an important role in the progression of ovarian serous cancers. The detection of DEK protein expression should be useful for the diagnosis and prognosis of ovarian serous cancers, and DEK might be a useful molecular target for ovarian cancer therapy. PMID- 19563408 TI - Fumagillin inhibits colorectal cancer growth and metastasis in mice: in vivo and in vitro study of anti-angiogenesis. AB - Fumagillin is an inhibitor of type 2 methionine aminopeptidase that can block blood vessel formation, but its molecular mechanism and therapeutic value in colon cancer still remain to be elucidated. In this study, male severe combined immunodeficiency (SCID) mice were injected with colon cancer cells in the subcutis and then treated with Fumagillin and Cyclo (Arg-Gly-Asp-D-Phe-Val), an integrin alphavbeta(3) antagonist. The tumor weight, microvessel density (MVD), and number of pulmonary metastatic foci were examined. Gene expression profiles were examined by microarray analysis of human umbilical endothelial cells (HUVEC). The Fumagillin-treated mice had smaller tumor mass, fewer pulmonary metastases, and lower MVD-CD105 levels than control animals. In vitro proliferation and tube formation of HUVEC was also significantly decreased by Fumagillin. Microarray analysis of Fumagillin-treated HUVEC showed upregulation of 71 genes and downregulation of 143 genes. Expression changes were involved in cell proliferation, migration, adhesion, and gene transcription. Quantitative real-time-polymerase chain reaction and western blotting showed decreased expression of cyclin E2, activated leukocyte cell adhesion molecule (ALCAM), and intercellular adhesion molecule-1 (ICAM-1) genes in the presence of Fumagillin. This downregulation by Fumagillin may be involved in the anti-angiogenesis by Fumagillin. In conclusion, Fumagillin was found to suppress colorectal cancer growth and metastasis by suppressing angiogenesis. PMID- 19563410 TI - Bile ductular cell reaction with senescent hepatocytes in chronic viral hepatitis is lost during hepatocarcinogenesis. AB - Cellular senescence is defined as irreversible cell arrest and could work as a safeguard against tumorigenesis. This mechanism was examined in chronic viral hepatitis-related hepatocarcinogenesis. By using surgical resected or wedge biopsied liver specimens from 87 chronic viral hepatitis patients in whom 35 neoplastic nodules (dysplastic nodules and hepatocellular carcinoma) were complicated, P21 expression and senescence-associated beta galactosidase activity, a marker of senescence, were examined. All of these neoplastic nodules harbored portal tracts within the tumors. Hepatocytes expressing senescence markers and cytokeratin (CK)7-positive bile ductules including hepatic progenitor like cells were increased in periseptal areas in cirrhosis. Interestingly, these cells appeared to form an anatomical complex that was completely lost in the periportal areas within the neoplastic nodules. In one-third of the neoplastic nodules, CK7-positive small neoplastic hepatocytes resembling hepatic progenitor cells proliferated zonally around the portal tracts. In conclusion, loss of a complex of senescent hepatocytes and ductular cell including hepatic progenitor like cells in the periportal or periseptal areas may be associated with emergence of neoplastic hepatocytes and their proliferation followed by neoplastic nodules arising in liver cirrhosis. Zonal proliferation of CK7-positive small neoplastic hepatocytes resembling hepatic progenitor cells may develop during early hepatocarcinogenesis. PMID- 19563409 TI - Immunohistochemical staining for P1 and P2 promoter-driven hepatocyte nuclear factor-4alpha may complement mucin phenotype of differentiated-type early gastric carcinoma. AB - Hepatocyte nuclear factor 4alpha (HNF4alpha) isoforms in the human stomach have not been fully investigated. The purpose of the present study was to evaluate the expression of P1 and P2 promoter-driven HNF4alpha (P1 and P2-HNF4alpha) in differentiated-type early gastric carcinomas (DEGC). P1- and P2-HNF4alpha expression was examined immunohistochemically both in non-neoplastic mucosa and carcinoma from surgical specimens. In all samples of non-neoplastic mucosa, foveolar, cardiac, fundic and pyloric gland epithelium was negative for P1 HNF4alpha, but was positive for P2-HNF4alpha. Intestinal metaplasia was positive for P1 and P2-HNF4alpha in all cases. Gastric carcinomas were classified into four mucin phenotypes based on the pattern of mucin expression: gastric, intestinal, mixed and null type. DEGC showed striking differences in the staining pattern for P1-HNF4alpha according to the mucin phenotype. Gastric carcinomas of intestinal, mixed and null type showed high positivity for P1-HNF4alpha, but the gastric type was negative for P1-HNF4alpha in all but one tumor. In contrast, P2 HNF4alpha was expressed in all tumors regardless of the mucin phenotype. Negative expression of P1-HNF4alpha was indicated as one of the useful immunohistochemical markers in the classification of mucin phenotype of both non-neoplastic mucosa and cancers of gastric phenotype. PMID- 19563411 TI - Bowel loop in an ovarian tumor: grossly visible, completely developed intestinal loop in mature cystic teratoma of malignant mixed germ cell tumor. AB - Although a gastrointestinal-type epithelium is observed in 7-13% of mature cystic teratoma cases, the occurrence of a grossly visible, organized gastrointestinal loop formation is very rare. Presented here is the case of a 14-year-old girl with malignant mixed germ cell tumor in the ovaries. In her left ovary a grossly visible, intestinal loop, 9 cm long, with hanging mesentery attached to the cystic wall of a mature cystic teratoma associated with a yolk sac tumor was observed, and in her right ovary another mature cystic teratoma was observed. Microscopy of the intestinal loop indicated a well-organized, intact layer of small intestinal wall. The yolk sac tumor predominantly had a poly-vitelline pattern. Previously, gastrointestinal wall or epithelium that was identified on microscopy has been reported. To the authors' knowledge this is the first case report of the formation of a grossly visible, completely developed intestinal loop in a malignant mixed germ cell tumor. PMID- 19563412 TI - Adenosquamous carcinoma in a congenital choledochal cyst associated with pancreatico-biliary maljunction. AB - Congenital choledochal cyst is occasionally complicated by carcinomatous transformation, mostly adenocarcinoma. Adenosquamous carcinoma arising in a congenital choledochal cyst is very rare. The author herein reports an adenosquamous carcinoma arising in congenital choledochal cyst associated with pancreatico-biliary maljunction. A 34-year-old man with congenital choledochal cyst and recurrent cholangitis had been followed up, and was admitted to hospital to undergo testing for cancer. Imaging modalities including computed tomography, magnetic resonance imaging and endoscopic retrograde cholangiography showed an elevated lesion in the choledochal cyst. Because clinical cytology of bile indicated malignant cells, pancreatico-duodenectomy, cholecystectomy, and resection of the choledochal cyst were performed. Grossly, the choledochal cyst was type I, and its size was 8 x 10 cm. Anomalous pancreatico-biliary ductal union was recognized. An elevated lesion was recognized in the choledochal cyst. Histologically, the lesion was composed of a squamous cell carcinoma element and an adenocarcinoma element; a gradual transition was recognized between the two. The squamous cell carcinoma element contained microcytic cells with mucins. On immunohistochemistry the adenocarcinoma element and microcytic cells were positive for CEA, but the squamous cell carcinoma element was negative for CEA. Both elements were positive for CA19-9. Ki-67 labeling was 53% in the adenocarcinoma element and 48% in the squamous cell carcinoma element. p53 protein was negative in both elements. At the time of writing, the patient was alive after 25 months without recurrence or metastasis. The present case is the second case of adenosquamous carcinoma arising in congenital choledochal cyst in the English-language literature. PMID- 19563413 TI - Adult-onset renal cell carcinoma associated with Xp11.2 translocations/TFE3 gene fusion with smooth muscle stroma and abnormal vessels. AB - Renal cell carcinoma (RCC) associated with Xp11.2 translocation/TFE3 gene fusion has been recently identified. Herein is presented a case of RCC with Xp11.2 translocations/TFE3 gene fusions with unusual histological findings. A 68-year old Japanese woman was incidentally found to have a renal mass on CT. Histological examination showed clear cell neoplasm with alveolar and papillary growth patterns. The nuclear atypia corresponded to Fuhrman grade 3. Additionally, smooth muscle stroma was observed and abnormal vessels showing a heterogeniety in thickness were also identified. On immunohistochemistry, neoplastic cells were diffusely positive for transcription factor E3 (TFE3) and Melan A, and focally positive for CD10 and RCC marker. The smooth muscle stroma was positive for alpha-smooth muscle actin and h-caldesmon, but reverse transcription-polymerase chain reaction of the tumor using frozen material could not detect any previously reported chimeric transcripts including ASPL-TFE3, PRCC TFE3, CLTC-TFE3, PSF-TFE3 or NoNo-TFE3. G-band karyotype was unsuccessful. Pathologists should pay attention to the afore-described unusual stromal reaction of adult-onset RCC associated with Xp11.2 translocations/TFE3 gene fusions. PMID- 19563415 TI - Volunteering pathology services and teaching in Ghana. PMID- 19563414 TI - Hepatocellular carcinoma occurring in a young Crohn's disease patient. AB - Reported herein is a case of hepatocellular carcinoma (HCC) occurring in a 25 year-old Japanese man who was diagnosed with Crohn's disease (CD) at 14 years of age; treatment included predonisolone, azathioprine, and infliximab. The tumor was located in right upper lobe and the size was 8 cm in diameter; histology was poorly differentiated HCC with pleomorphic cellular changes. Adjacent normal liver showed no evidence of cirrhosis or viral hepatitis. Until now, only six cases of HCC arising in patients with CD have been reported in the English language literature. Most of these patients had early onset of CD and HCC: none had cirrhosis or virus hepatitis. Most patients had a long disease history of CD and were being medicated with several immunosuppressive agents. Some factors associated with CD might indirectly or directly be related to the development of HCC in CD patients, although the possibility that these HCC occurred coincidentally in CD patients, including the present patient, cannot be ruled out. Accumulation of cases is necessary to evaluate the relationship between CD and HCC precisely. PMID- 19563416 TI - Actual Proliferating Index and p53 protein expression as prognostic marker in odontogenic cysts. AB - BACKGROUND: The purpose of this study was to evaluate the biological aggressiveness of odontogenic keratocyst/keratocystic odontogenic tumour (KCOT), radicular cyst (RC) and dentigerous cyst (DC) by observing the actual proliferative activity of epithelium, and p53 protein expression. METHODS: The actual proliferative activity was measured by Ki-67 Labelling Index and argyrophilic nucleolar organizing regions (AgNOR) count per nucleus. The p53 protein expression was also evaluated. RESULTS: Ki-67 positive cells were observed higher in suprabasal cell layers of KCOT with uniform distribution, a few of them were predominantly observed in basal cell layer in RC and DC. The AgNOR count was significantly higher in suprabasal cell layers of KCOT. The actual proliferative activity was noted to be higher in suprabasal cell layers of KCOT. The p53 immunolabelling was dense and scattered in basal and suprabasal cell layers in KCOT. The weakly stained p53 positive cells were observed diffusely distributed in KCOT, whereas they were mainly seen in basal cell layer of RC and DC. CONCLUSION: The quantitative and qualitative differences of the proliferative activity and the p53 protein expression in sporadic KCOT may be associated with intrinsic growth potential that could play a role in its development and explain locally aggressive biological behaviour. AgNOR count and p53 protein detection in odontogenic lesions can be of great consequence to predict the biological behaviour and prognosis. PMID- 19563417 TI - Humoral immunity to cow's milk proteins and gliadin within the etiology of recurrent aphthous ulcers? AB - OBJECTIVES: The goal of this study was to determine the incidence of serum antibodies to gliadin and to cow's milk proteins (CMP) using ELISA test, within patients who have recurrent aphthous ulcers (RAU). SUBJECTS AND METHODS: Fifty patients with recurrent aphthous ulcers and fifty healthy people were included in this research. Levels of serum IgA and IgG antibodies to gliadin and IgA, IgG and IgE to CMP were determined using ELISA. RESULTS: The levels of serum antigliadin IgA and IgG antibodies were not significantly higher in patients with RAU in comparison with the controls (P = 0.937 and P = 0.1854 respectively). The levels of serum anti-CMP IgA, IgG and IgE antibodies were significantly higher in patients with RAU in comparison with the controls (P < 0.005, P < 0.002 and P < 0.001 respectively). In general, the increased humoral (IgA or IgG) immunoreactivity to CMP was found in 32 of 50 patients, while 17 of them showed the increased levels of both IgA and IgG immunoreactivity to CMP. At the same time, 16 out of 50 patients had IgA, IgG and IgE immunoreactivity to CMP. CONCLUSION: These results indicate the strong association between high levels of serum anti-CMP IgA, IgG and IgE antibodies and clinical manifestations of recurrent aphthous ulcers. PMID- 19563418 TI - Serum soluble CD44v6 levels in patients with oral and maxillofacial malignancy. AB - OBJECTIVE: To determine the levels of serum sCD44v6 in patients with oral cancer and evaluate the value of serum sCD44v6 in adjuvant diagnosis, staging and monitoring treatment response in these patients. MATERIALS AND METHODS: A total of 112 hospitalized patients with oral and maxillofacial malignancy and 28 healthy individuals were examined for serum sCD44v6 levels. Venous blood was collected from these patients and the healthy individuals. One week after treatment, venous blood was collected once again in 60 patients with oral and maxillofacial squamous cell carcinoma (OSCC). RESULTS: The sCD44v6 concentration was not significantly different between patients with oral and maxillofacial malignancy and control group (P > 0.05). The levels of serum sCD44v6 in patients with OSCC and salivary carcinoma showed no difference with those in control group (P > 0.05). The sCD44v6 level in patients with stage III and IV disease was higher than that of patients with stage I and II and that of the control group, but the difference was not significant (P > 0.05). Serum sCD44v6 levels in patients with OSCC after treatment became lower than that prevailed during pretreatment (P < 0.05). CONCLUSION: The possible roles of CD44v6 in the diagnosis of oral and maxillofacial malignancy deserve further elucidation and evaluation. Serum sCD44v6 may be a valuable marker in monitoring treatment response in patients with OSCC. PMID- 19563419 TI - Solute and heat transport model of the Henry and hilleke laboratory experiment. AB - SEAWAT is a coupled version of MODFLOW and MT3DMS designed to simulate variable density ground water flow and solute transport. The most recent version of SEAWAT, called SEAWAT Version 4, includes new capabilities to represent simultaneous multispecies solute and heat transport. To test the new features in SEAWAT, the laboratory experiment of Henry and Hilleke (1972) was simulated. Henry and Hilleke used warm fresh water to recharge a large sand-filled glass tank. A cold salt water boundary was represented on one side. Adjustable heating pads were used to heat the bottom and left sides of the tank. In the laboratory experiment, Henry and Hilleke observed both salt water and fresh water flow systems separated by a narrow transition zone. After minor tuning of several input parameters with a parameter estimation program, results from the SEAWAT simulation show good agreement with the experiment. SEAWAT results suggest that heat loss to the room was more than expected by Henry and Hilleke, and that multiple thermal convection cells are the likely cause of the widened transition zone near the hot end of the tank. Other computer programs with similar capabilities may benefit from benchmark testing with the Henry and Hilleke laboratory experiment. PMID- 19563420 TI - Estimating flow using tracers and hydraulics in synthetic heterogeneous aquifers. AB - Regional ground water flow is most usually estimated using Darcy's law, with hydraulic conductivities estimated from pumping tests, but can also be estimated using ground water residence times derived from radioactive tracers. The two methods agree reasonably well in relatively homogeneous aquifers but it is not clear which is likely to produce more reliable estimates of ground water flow rates in heterogeneous systems. The aim of this paper is to compare bias and uncertainty of tracer and hydraulic approaches to assess ground water flow in heterogeneous aquifers. Synthetic two-dimensional aquifers with different levels of heterogeneity (correlation lengths, variances) are used to simulate ground water flow, pumping tests, and transport of radioactive tracers. Results show that bias and uncertainty of flow rates increase with the variance of the hydraulic conductivity for both methods. The bias resulting from the nonlinearity of the concentration-time relationship can be reduced by choosing a tracer with a decay rate similar to the mean ground water residence time. The bias on flow rates estimated from pumping tests is reduced when performing long duration tests. The uncertainty on ground water flow is minimized when the sampling volume is large compared to the correlation length. For tracers, the uncertainty is related to the ratio of correlation length to the distance between sampling wells. For pumping tests, it is related to the ratio of correlation length to the pumping test's radius of influence. In regional systems, it may be easier to minimize this ratio for tracers than for pumping tests. PMID- 19563421 TI - Global optimal design of ground water monitoring network using embedded kriging. AB - We present a methodology for global optimal design of ground water quality monitoring networks using a linear mixed-integer formulation. The proposed methodology incorporates ordinary kriging (OK) within the decision model formulation for spatial estimation of contaminant concentration values. Different monitoring network design models incorporating concentration estimation error, variance estimation error, mass estimation error, error in locating plume centroid, and spatial coverage of the designed network are developed. A big-M technique is used for reformulating the monitoring network design model to a linear decision model while incorporating different objectives and OK equations. Global optimality of the solutions obtained for the monitoring network design can be ensured due to the linear mixed-integer programming formulations proposed. Performances of the proposed models are evaluated for both field and hypothetical illustrative systems. Evaluation results indicate that the proposed methodology performs satisfactorily. These performance evaluation results demonstrate the potential applicability of the proposed methodology for optimal ground water contaminant monitoring network design. PMID- 19563422 TI - Variable-density flow and transport simulation of wellbore brine displacement. AB - Borehole dilution tests have been used for characterization of aquifer hydrogeologic properties for several decades. Based on the principles of borehole dilution tests, we conducted what more appropriately may be considered a wellbore fluid displacement test in a limestone aquifer in South Carolina. Our study area is a quarry in the coastal plain of South Carolina. Using a solution of reagent grade NaCl and deionized H(2)O as a tracer, a brine slug was introduced into a 5 cm (2 in.) diameter Schedule 40 PVC well with a 6-m slotted screen at the bottom. Immediately following addition of the brine, a recording electrical conductivity (EC) sensor was placed in the well opposite the screen and set to record EC in 2 min intervals for 5 days. An alternative to previous methods for analyzing data from wellbore brine displacement tests was developed. Results were analyzed using SEAWAT-2000 to account for the density dependency of brine flow and transport. The high spatial resolution, three-dimensional numerical simulation enabled direct incorporation of well construction peculiarities, including the sand pack and length of screen, in the data analysis. Hydraulic conductivity, effective porosity, and longitudinal dispersivity were adjusted in the simulation model until the best match of simulated wellbore fluid concentrations to observed concentrations was achieved. Using this procedure, we were able to obtain a very close agreement between observed and simulated concentrations and, hence, reliable estimates of the hydrogeologic properties of the aquifer in the vicinity of the test well. PMID- 19563423 TI - Another look at steady-shape conditions. PMID- 19563424 TI - Eolian transport of geogenic hexavalent chromium to ground water. AB - A conceptual model of eolian transport is proposed to address the widely distributed, high concentrations of hexavalent chromium (Cr(+6)) observed in ground water in the Emirate of Abu Dhabi, United Arab Emirates. Concentrations (30 to more than 1000 microg/L Cr(+6)) extend over thousands of square kilometers of ground water systems. It is hypothesized that the Cr is derived from weathering of chromium-rich pyroxenes and olivines present in ophiolite sequence of the adjacent Oman (Hajar) Mountains. Cr(+3) in the minerals is oxidized to Cr(+6) by reduction of manganese and is subsequently sorbed on iron and manganese oxide coatings of particles. When the surfaces of these particles are abraded in this arid environment, they release fine, micrometer-sized, coated particles that are easily transported over large distances by wind and subsequently deposited on the surface. During ground water recharge events, the readily soluble Cr(+6) is mobilized by rain water and transported by advective flow into the underlying aquifer. Chromium analyses of ground water, rain, dust, and surface (soil) deposits are consistent with this model, as are electron probe analyses of clasts derived from the eroding Oman ophiolite sequence. Ground water recharge flux is proposed to exercise some control over Cr(+6) concentration in the aquifer. PMID- 19563425 TI - Practical modeling approaches for geological storage of carbon dioxide. AB - The relentless increase of anthropogenic carbon dioxide emissions and the associated concerns about climate change have motivated new ideas about carbon constrained energy production. One technological approach to control carbon dioxide emissions is carbon capture and storage, or CCS. The underlying idea of CCS is to capture the carbon before it emitted to the atmosphere and store it somewhere other than the atmosphere. Currently, the most attractive option for large-scale storage is in deep geological formations, including deep saline aquifers. Many physical and chemical processes can affect the fate of the injected CO2, with the overall mathematical description of the complete system becoming very complex. Our approach to the problem has been to reduce complexity as much as possible, so that we can focus on the few truly important questions about the injected CO2, most of which involve leakage out of the injection formation. Toward this end, we have established a set of simplifying assumptions that allow us to derive simplified models, which can be solved numerically or, for the most simplified cases, analytically. These simplified models allow calculation of solutions to large-scale injection and leakage problems in ways that traditional multicomponent multiphase simulators cannot. Such simplified models provide important tools for system analysis, screening calculations, and overall risk-assessment calculations. We believe this is a practical and important approach to model geological storage of carbon dioxide. It also serves as an example of how complex systems can be simplified while retaining the essential physics of the problem. PMID- 19563426 TI - Analytical design curves to maximize pumping or minimize injection in coastal aquifers. AB - Explicit algebraic equations are derived to determine approximate maximum pumping rates or minimum injection rates to limit sea water intrusion to a prespecified distance from the coastline. The equations are based on Strack's (1976) single potential solution. The maximum pumping rates and minimum injection rates applied at wells with uniform spacing to control the inland movement of the fresh water salt water interface in a coastal aquifer could be calculated from Strack's (1976) solution without the need of a numerical optimization algorithm. When wells are distributed in a simple fashion, the maximum intrusion location can be identified precisely for pumping cases and approximately for injection cases. For pumping cases, critical points are the limit of allowable salt water intrusion, whereas no such limit exists for injection cases. Once an application site is identified, a series of design curves for pumping and injection rates can be developed for arbitrary intrusion limits. When a user is interested only in the largest pumping rates associated with critical points, one design curve can yield complete information. PMID- 19563427 TI - A parallel PCG solver for MODFLOW. AB - In order to simulate large-scale ground water flow problems more efficiently with MODFLOW, the OpenMP programming paradigm was used to parallelize the preconditioned conjugate-gradient (PCG) solver with in this study. Incremental parallelization, the significant advantage supported by OpenMP on a shared-memory computer, made the solver transit to a parallel program smoothly one block of code at a time. The parallel PCG solver, suitable for both MODFLOW-2000 and MODFLOW-2005, is verified using an 8-processor computer. Both the impact of compilers and different model domain sizes were considered in the numerical experiments. Based on the timing results, execution times using the parallel PCG solver are typically about 1.40 to 5.31 times faster than those using the serial one. In addition, the simulation results are the exact same as the original PCG solver, because the majority of serial codes were not changed. It is worth noting that this parallelizing approach reduces cost in terms of software maintenance because only a single source PCG solver code needs to be maintained in the MODFLOW source tree. PMID- 19563429 TI - Evaluation of cluster-randomized trials on maternal and child health research in developing countries. AB - OBJECTIVE: To summarize and evaluate all publications including cluster randomized trials used for maternal and child health research in developing countries during the last 10 years. METHODS: All cluster-randomized trials published between 1998 and 2008 were reviewed, and those that met our criteria for inclusion were evaluated further. The criteria for inclusion were that the trial should have been conducted in maternal and child health care in a developing country and that the conclusions should have been made on an individual level. Methods of accounting for clustering in design and analysis were evaluated in the eligible trials. RESULTS: Thirty-five eligible trials were identified. The majority of them were conducted in Asia, used community as randomization unit, and had less than 10,000 participants. To minimize confounding, 23 of the 35 trials had stratified, blocked, or paired the clusters before they were randomized, while 17 had adjusted for confounding in the analysis. Ten of the 35 trials did not account for clustering in sample size calculations, and seven did not account for the cluster-randomized design in the analysis. The number of cluster-randomized trials increased over time, and the trials generally improved in quality. CONCLUSIONS: Shortcomings exist in the sample-size calculations and in the analysis of cluster-randomized trials conducted during maternal and child health research in developing countries. Even though there has been improvement over time, further progress in the way that researchers utilize and analyse cluster-randomized trials in this field is needed. PMID- 19563428 TI - Nasopharyngeal carriage of S. pneumoniae among young children in rural Nepal. AB - OBJECTIVES: To provide epidemiologic data on Streptococcus pneumoniae (Spn) carriage in Nepal. METHODS: Prospective, population-based study among children in Sarlahi, Nepal to estimate carriage prevalence, identify risk factors, and determine antibiotic susceptibility patterns and serotype distribution. Between December 2003 and July 2004, NP specimens were collected from 604 children aged 1 36 months with acute lower respiratory infection (ALRI) and 604 healthy, age- and season-matched controls. RESULTS: Of the 1100 specimens analysed, carriage prevalence was approximately 80% in both groups. In the multivariate analyses, significant risk factors for Spn carriage in controls were Muslim religion [adjusted odds ratio (AOR): 2.93] and no latrine in the household (AOR: 2.41). Those treated for a recent illness had lower carriage rates (AOR: 0.37). Results were similar for ALRI cases with the addition of age >or=12 months (AOR: 1.68), and symptomatic infection (AOR: 3.78) as risk factors. The antibiotics and proportions of isolates resistant to them were as follows: penicillin 4.5%, cotrimoxazole 89.2%, chloramphenicol 1.4%, erythromycin 1.5% and tetracycline 22.7%. The most prevalent serogroups/types were 6, 19, 23, 15, 9 and 10. CONCLUSIONS: Young children in rural Nepal experience high rates of Spn carriage. Most isolates were resistant to cotrimoxazole. Current conjugate Spn vaccines may substantially reduce the risk of a severe pneumonia and other Spn infections. PMID- 19563430 TI - Spatial distribution and risk factors of dengue and Japanese encephalitis virus infection in urban settings: the case of Vientiane, Lao PDR. AB - OBJECTIVES: To evaluate the prevalence of flavivirus infection in Vientiane city (Lao PDR), to describe the spatial distribution of infection within this city, and to explore the link between flavivirus seroprevalence and urbanization levels of residential neighbourhoods. METHODS: A seroprevalence survey was carried out in 2006 including 1990 adults (>or=35 years) and 1568 children (>or=6 months and <6 years) randomly selected. RESULTS: The prevalence of individuals with previous flavivirus infection (i.e. negative for both DEN and JE IgM but positive for DEN IgG) was 57.7%, with a significantly (P < 0.001) higher prevalence among adults (84.6%; 95% confidence interval (CI) = 82.4-86.8) than children (9.4%; 95% CI = 7.2-11.6). The prevalence of individuals with recent flavivirus infection (i.e. positive for DEN and/or JE IgM) was 6.5% and also significantly (P < 0.001) higher among adults (10.0%; 95% CI = 8.3-11.7) than children (2.5%; 95% CI = 1.5 3.5). In terms of spatial distribution, IgG prevalence was significantly (P < 0.001) higher among individuals living in the central city (60.1%; 95% CI = 56.2 64.1) than among those living in the periphery (44.3%; 95% CI = 41.5-47.2). In contrast, seroprevalence of recent flavivirus infections was significantly (P < 0.001) higher among individuals living in the periphery (8.8%; 95% CI = 6.9-10.7) than in the central city (4.0%; 95% CI = 2.9-5.2). This association was also statistically consistent (P < 0.01) in multivariate logistic regression after controlling for individual risk factors. CONCLUSIONS: Our findings indicate that the level of urbanization of residential neighbourhoods influences the risk of flavivirus infection. The spatial distribution of flavivirus infection varies, even within a small city of less than 300,000 habitants such as Vientiane. PMID- 19563432 TI - Tuberculosis cluster in an immigrant community: case identification issues and a transcultural perspective. AB - OBJECTIVES: In a low incidence area for tuberculosis (TB), a computerized database identified an unusually high proportion of patients coming from one single country between 2004 and 2006. To determine whether they constituted a cluster, whether clustering was due to recent transmission, and to understand what undermined the efficacy of the contact tracing procedure, we conducted a retrospective study of all patients with TB from this country. METHODS: Mycobacterium tuberculosis isolates of 15 TB cases originating from the same country over a 2(1/2) year period were analysed by restriction fragment length polymorphism (RFLP) and/or Rep-PCR. To identify links between patients, we revisited the social worker's files, cross-matched contacts' databases, and performed internet searches. A cultural evaluation was conducted by an anthropologist and an expert physician, through patient and community key informant interviews and a literature review. RESULTS: Genotyping confirmed that 11 of 15 patients had identical isolates. Additional data revealed an unsuspected complex network of social links between 9 of these 11 patients. The transcultural evaluation pointed out the major obstacles to efficient contact tracing, such as importance of social stigma related to TB, differences in communication style and health beliefs, and linguistic barriers. CONCLUSION: The combined finding of identical genotypes and important social links between patients confirmed the suspicion of a TB cluster due to recent transmission. The cultural evaluation helped to explain the difficulties encountered during the contact tracing procedure, and offered strategies to improve its efficacy despite the magnitude of the social stigma attached to TB in this community. PMID- 19563431 TI - Predictors for mortality and loss to follow-up among children receiving anti retroviral therapy in Lilongwe, Malawi. AB - OBJECTIVES: To determine predictors of mortality in children on anti-retroviral therapy (ART) who attended the Paediatric HIV Clinic at Kamuzu Central Hospital in Lilongwe, Malawi. METHODS: Retrospective case cohort study by chart review of children who had started ART between October 2004 and May 2006. Bivariable and multivariable analysis were performed with and without defaulters to evaluate associations according to vital status and to identify independent predictors of mortality. RESULTS: Forty-one of 258 children (15.9%) were deceased, 185 (71.7%) were alive, and 32 (12.4%) had defaulted: 51% were female, 7% were under 18 months, 26% were 18 months to 5 years, and 54% were >5 years of age. Most were WHO stage III or IV (56% and 37%, respectively). On multivariate analysis, factors most strongly associated with mortality and defaulting were age <18 months [hazards ratio (HR) 2.11 (95% CI 1.0-4.51)] and WHO stage IV [HR 2.00 (95% CI 1.07-3.76)]. CONCLUSIONS: To improve outcomes of HIV-positive children, they must be identified and treated early, specifically children under 18 months of age. Access to infant diagnostic procedures must be improved to allow effective initiation of ART in infants at higher risk of death. PMID- 19563433 TI - Re-emerging chikungunya fever: some lessons from Asia. AB - Chikungunya is an arboviral infection that re-emerged in several Asian countries during 2005-2006 after a long period of quiescence. Several microbial, climatic, social and economic factors influenced the occurrence of this disease as well as the rapidity with which it swept across many countries resulting in significant morbidity. Prevention and control of such diseases require not only a strong public health infrastructure but also a precise understanding of the factors that provide a conducive environment for the virus to propagate and infect a large number of people in a short time period. A multipronged response with an active role by the communities is critical for combating chikungunya and other emerging infectious diseases. The paper discusses important lessons that can be learned from the recent outbreaks of chikungunya fever in Asia. PMID- 19563434 TI - Cost-effectiveness projections of single and combination therapies for visceral leishmaniasis in Bihar, India. AB - OBJECTIVES: To assess the cost-effectiveness of current monotherapies and prospective combinations for treating visceral leishmaniasis (VL) in Bihar, India in terms of years of life lost (YLL) averted as well as deaths averted. METHODS: We employed two methods to estimate the number of avertable deaths in our analysis: one using estimated mortality, the other using direct incidence estimates for VL. Costs of care are based on an average private hospital in Bihar, and data on drug costs were obtained both locally and from the World Health Organization. RESULTS: The cost of monotherapies per averted YLL ranged from US$2 for paromomycin in an outpatient setting to US$20-22 for AmBisome at 20 mg/kg. The corresponding costs per death averted ranged from US$53-54 to US$523 527. Combinations ranged US$5-8 per YLL averted and US$124-213 per death averted. CONCLUSION: The available treatments for VL are cost-effective, and our mortality and incidence-based methods produce consistent estimates. The combinations considered here were more cost-effective than most monotherapies. Having multiple treatment options and combining drugs are also likely to reduce drug pressure and prolong the drugs' life-span of effective use. PMID- 19563435 TI - A mutation in the Arabidopsis mTERF-related plastid protein SOLDAT10 activates retrograde signaling and suppresses (1)O(2)-induced cell death. AB - The conditional flu mutant of Arabidopsis thaliana generates singlet oxygen ((1)O(2)) in plastids during a dark-to-light shift. Seedlings of flu bleach and die, whereas mature plants stop growing and develop macroscopic necrotic lesions. Several suppressor mutants, dubbed singlet oxygen-linked death activator (soldat), were identified that abrogate (1)O(2)-mediated cell death of flu seedlings. One of the soldat mutations, soldat10, affects a gene encoding a plastid-localized protein related to the human mitochondrial transcription termination factor mTERF. As a consequence of this mutation, plastid-specific rRNA levels decrease and protein synthesis in plastids of soldat10 is attenuated. This disruption of chloroplast homeostasis in soldat10 seedlings affects communication between chloroplasts and the nucleus and leads to changes in the steady-state concentration of nuclear gene transcripts. The soldat10 seedlings suffer from mild photo-oxidative stress, as indicated by the constitutive up regulation of stress-related genes. Even though soldat10/flu seedlings overaccumulate the photosensitizer protochlorophyllide in the dark and activate the expression of (1)O(2)-responsive genes after a dark-to-light shift they do not show a (1)O(2)-dependent cell death response. Disturbance of chloroplast homeostasis in emerging soldat10/flu seedlings seems to antagonize a subsequent (1)O(2)-mediated cell death response without suppressing (1)O(2)-dependent retrograde signaling. The results of this work reveal the unexpected complexity of what is commonly referred to as 'plastid signaling'. PMID- 19563436 TI - Phosphorylation at S384 regulates the activity of the TaALMT1 malate transporter that underlies aluminum resistance in wheat. AB - In this study we examined the role of protein phosphorylation/dephosphorylation in the transport properties of the wheat (Triticum aestivum) root malate efflux transporter underlying Al resistance, TaALMT1. Pre-incubation of Xenopus laevis oocytes expressing TaALMT1 with protein kinase inhibitors (K252a and staurosporine) strongly inhibited both basal and Al(3+)-enhanced TaALMT1-mediated inward currents (malate efflux). Pre-incubation with phosphatase inhibitors (okadaic acid and cyclosporine A) resulted in a modest inhibition of the TaALMT1 mediated currents. Exposure to the protein kinase C (PKC) activator, phorbol 12 myristate 13-acetate (PMA), enhanced TaALMT1-mediated inward currents. Since these observations suggest that TaALMT1 transport activity is regulated by PKC mediated phosphorylation, we proceeded to modify candidate amino acids in the TaALMT1 protein in an effort to identify structural motifs underlying the process regulating phosphorylation. The transport properties of eight single point mutations (S56A, S183A, S324A, S337A, S351-352A, S384A, T323A and Y184F) generated in amino acid residues predicted to be phosphorylation sites and examined electrophysiologically. The basic transport properties of mutants S56A, S183A, S324A, S337A, S351-352A, T323A and Y184F were not altered relative to the wild-type TaALMT1. Likewise the sensitivity of these mutants to staurosporine resembled that observed for the wild-type transporter. However, the mutation S384A was noticeable, as in oocytes expressing this mutant protein TaALMT1 mediated basal and Al-enhanced currents were significantly inhibited, and the currents were insensitive to staurosporine or PMA. These findings indicate that S384 is an essential residue regulating TaALMT1 activity via direct protein phosphorylation, which precedes Al(3+) enhancement of transport activity. PMID- 19563437 TI - Uracil salvage is necessary for early Arabidopsis development. AB - Uridine nucleotides can be formed by energy-consuming de novo synthesis or by the energy-saving recycling of nucleobases resulting from nucleotide catabolism. Uracil phosphoribosyltransferases (UPRTs; EC 2.4.2.9) are involved in the salvage of pyrimidines by catalyzing the formation of uridine monophosphate (UMP) from uracil and phosphoribosylpyrophosphate. To date, UPRTs are described as non essential, energy-saving enzymes. In the present work, the six genes annotated as UPRTs in the Arabidopsis genome are examined through phylogenetic and functional complementation approaches and the available T-DNA insertion mutants are characterized. We show that a single nuclear gene encoding a protein targeted to plastids, UPP, is responsible for almost all UPRT activity in Arabidopsis. The inability to salvage uracil caused a light-dependent dramatic pale-green to albino phenotype, dwarfism and the inability to produce viable progeny in loss-of function mutants. Plastid biogenesis and starch accumulation were affected in all analysed tissues, with the exception of stomata. Therefore we propose that uracil salvage is of major importance for plant development. PMID- 19563438 TI - Interaction between the light quality and flowering time pathways in Arabidopsis. AB - Flowering in Arabidopsis is accelerated by a reduced ratio of red light to far red light (R/FR), which indicates the proximity of competitive vegetation. By exploiting the natural genetic variation in flowering time responses to low R/FR, we obtained further insight into the complex pathways that fine-tune the transition to flowering in Arabidopsis. The Bla-6 ecotype does not flower significantly earlier in response to low R/FR, but is still able to display other features of shade avoidance, suggesting branching of low R/FR signalling. Here we show that the muted flowering response of Bla-6 is due to high levels of the floral repressor FLOWERING LOCUS C (FLC), conferred by a combination of functional FLC and FRIGIDA (FRI) alleles with a 'weak'FY allele. The Bla-6 FY allele encodes a protein with a corrupted WW binding domain, and we provide evidence that this locus plays a key role in the natural variation in light quality-induced flowering in Arabidopsis. In Bla-6, FLC blocks promotion to flowering by reduced R/FR by inhibiting expression of the floral integrator FLOWERING LOCUS T (FT) in a dose-dependent manner. Reduction of FLC removes this obstruction, and Bla6 plants then exhibit strong induction of FT and flower early in response to a low R/FR signal. This paper illustrates the intricate interaction of environmental signals and genetic factors to regulate flowering in Arabidopsis. PMID- 19563439 TI - Three-dimensional imaging of plant cuticle architecture using confocal scanning laser microscopy. AB - Full appreciation of the roles of the plant cuticle in numerous aspects of physiology and development requires a comprehensive understanding of its biosynthesis and deposition; however, much is still not known about cuticle structure, trafficking and assembly. To date, assessment of cuticle organization has been dominated by 2D imaging, using histochemical stains in conjunction with light and fluorescence microscopy. This strategy, while providing valuable information, has limitations because it attempts to describe a complex 3D structure in 2D. An imaging technique that could accurately resolve 3D architecture would provide valuable additions to the growing body of information on cuticle molecular biology and biochemistry. We present a novel application of 3D confocal scanning laser microscopy for visualizing the architecture, deposition patterns and micro-structure of plant cuticles, using the fluorescent stain auramine O. We demonstrate the utility of this technique by contrasting the fruit cuticle of wild-type tomato (Solanum lycopersicum cv. M82) with those of cutin-deficient mutants. We also introduce 3D cuticle modeling based on reconstruction of serial optical sections, and describe its use in identification of several previously unreported features of the tomato fruit cuticle. PMID- 19563440 TI - A moderate decrease in temperature induces COR15a expression through the CBF signaling cascade and enhances freezing tolerance. AB - Temperature has a profound effect on plant growth and development. However, the molecular mechanisms underlying this regulation are not well understood. In particular, how moderate temperature variations are perceived and transduced inside the plant cells remains obscure. In this study, we analyzed transcriptional responses to a moderate decrease in temperature (cooling) in Arabidopsis thaliana. The cooling response involves a weaker and more transient induction of cold-induced genes, such as COR15a, than cold response. This induction probably accounts for the increase in freezing tolerance by cooling acclimation. Cooling also induces some defense response genes, and their induction, but not that of COR15a, requires the salicylic acid signaling pathway. Analysis of the regulation of COR15a reveals that cooling induction is mediated through the same C repeat/dehydration-responsive (CRT/DRE) element as cold induction. Furthermore, we identified a role for CBF1 and CBF4 in transducing signals of moderate decreases in temperature. It appears that variants of the CBF signaling cascade are utilized in cold and cooling responses, and a moderate decrease in temperature may invoke an adaptive response to prepare plants to cope with a more drastic decrease in temperature. PMID- 19563441 TI - Heterogeneity of Arabidopsis core promoters revealed by high-density TSS analysis. AB - Our limited understanding of plant promoters does not allow us to recognize any core promoter elements for the majority of plant promoters. To understand the promoter architecture of Arabidopsis, we used the combined approach of in silico detection of novel core promoter elements and large-scale determination of transcription start sites (TSSs). To this end, we developed a novel methodology for TSS identification, using a combination of the cap-trapper and massively parallel signature sequencing methods. This technique, CT-MPSS, allowed us to identify 158 237 Arabidopsis TSS tags corresponding to 38 311 TSS loci, which provides an opportunity for quantitative analysis of plant promoters. The expression characteristics of these promoters were analyzed with respect to core promoter elements detected by our in silico analyses, revealing that Arabidopsis promoters contain two main types of elements with exclusive characteristics, the TATA type and the GA type. The TATA-type promoters tend to be associated with the Y Patch and the Inr motif, and cause high expression with sharp-peak TSS clusters. By contrast, the GA type produces broad-type TSS clusters. Unlike mammalian promoters, plant promoters are not associated with CpG islands. However, plant-specific GA-type promoters share some characteristics with mammalian CpG-type promoters. PMID- 19563442 TI - Dissecting the regulation of fructan metabolism in chicory (Cichorium intybus) hairy roots. AB - Fifteen per cent of higher plants accumulate fructans. Plant development, nutritional status and stress exposure all affect fructan metabolism, and while fructan biochemistry is well understood, knowledge of its regulation has remained fragmentary. Here, we have explored chicory (Cichorium intybus) hairy root cultures (HRCs) to study the regulation of fructan metabolism in sink tissues in response to environmental cues. In standard medium (SM), HRCs did not accumulate inulin. However, upon transfer to high-carbon (C)/low-nitrogen (N) medium, expression of sucrose:sucrose 1-fructosyltransferase (1-SST) and fructan:fructan 1-fructosyltransferase (1-FFT) was strongly induced and inulin accumulated. Upon return to SM, inulin was degraded, together with a coordinate decline of 1-SST and 1-FFT expression. In HRCs, cold-induced expression of fructan 1-exohydrolases (1-FEH I and IIa) was similar to cold induction in taproots, even in the absence of accumulated inulin. For high-C/low-N induction of 1-SST and 1-FFT, and cold induction of 1-FEH I and IIa, the signaling pathways were addressed. While 1-SST and 1-FFT induction was similarly prevented by inhibitors of Ca(2+) signaling, protein kinases and phosphatases, cold induction of 1-FEH I and IIa revealed distinct signaling pathways. In summary, this study has established chicory HRCs as a convenient experimental system with which to study the regulation of fructan active enzyme (FAZY) expression in heterotrophic cells. PMID- 19563443 TI - Functional repair of embolized vessels in maize roots after temporal drought stress, as demonstrated by magnetic resonance imaging. AB - Xylem sap under high tension is in a metastable state and tends to cavitate, frequently leading to an interruption of the continuous water columns. Mechanisms of cavitation repair are controversially discussed. Magnetic resonance (MR) imaging provides a noninvasive, high spatial and temporal resolution approach to monitor xylem cavitation, refilling, and functionality. Spin density maps of drought-stressed maize taproots were recorded to localize cavitation events and to visualize the refilling processes; c. 2 h after release of the nutrient solution from the homemade MR imaging cuvette that received the root, late metaxylem vessels started to cavitate randomly as identified by a loss of signal intensity. After c. 6 h plants were rewatered, leading to a repair of water columns in five out of eight roots. Sap ascent during refilling, monitored with multislice MR imaging sequences, varied between 0.5 mm min(-1) and 3.3 mm min( 1). Flow imaging of apparently refilled vessels was performed to test for functional repair. Occasionally, a collapse of xylem vessels under tension was observed; this collapse was reversible upon rewatering. Refilling was an all-or none process only observed under low-light conditions. Absence of flow in some of the apparently refilled vessels indicates that functionality was not restored in these particular vessels, despite a recovery of the spin density signal. PMID- 19563444 TI - Global patterns of foliar nitrogen isotopes and their relationships with climate, mycorrhizal fungi, foliar nutrient concentrations, and nitrogen availability. AB - Ratios of nitrogen (N) isotopes in leaves could elucidate underlying patterns of N cycling across ecological gradients. To better understand global-scale patterns of N cycling, we compiled data on foliar N isotope ratios (delta(15)N), foliar N concentrations, mycorrhizal type and climate for over 11,000 plants worldwide. Arbuscular mycorrhizal, ectomycorrhizal, and ericoid mycorrhizal plants were depleted in foliar delta(15)N by 2 per thousand, 3.2 per thousand, 5.9 per thousand, respectively, relative to nonmycorrhizal plants. Foliar delta(15)N increased with decreasing mean annual precipitation and with increasing mean annual temperature (MAT) across sites with MAT >or= -0.5 degrees C, but was invariant with MAT across sites with MAT < -0.5 degrees C. In independent landscape-level to regional-level studies, foliar delta(15)N increased with increasing N availability; at the global scale, foliar delta(15)N increased with increasing foliar N concentrations and decreasing foliar phosphorus (P) concentrations. Together, these results suggest that warm, dry ecosystems have the highest N availability, while plants with high N concentrations, on average, occupy sites with higher N availability than plants with low N concentrations. Global-scale comparisons of other components of the N cycle are still required for better mechanistic understanding of the determinants of variation in foliar delta(15)N and ultimately global patterns in N cycling. PMID- 19563445 TI - Prevalence of metabolic syndrome and its relationship with leisure time physical activity among Peruvian adults. AB - BACKGROUND: Metabolic syndrome (MetS) is an important risk factor of cardiovascular disease (CVD) and type 2 diabetes. Previous studies have suggested an inverse relationship between physical activity and MetS. However, these findings were inconsistent, and few investigators have examined these associations among South Americans. We estimated the prevalence of MetS and its association with leisure time physical activity (LTPA) among Peruvian adults. MATERIALS AND METHODS: This cross-sectional study of 1675 individuals (619 men and 1056 women) was conducted among residents in Lima and Callao, Peru. Information about LTPA, socio-demographical and other lifestyle characteristics was collected by interview. The presence of MetS was defined using the Adult Treatment Panel III criteria. RESULTS: Overall, the prevalence of MetS was 26.9% and was more common among women (29.9%) than men (21.6%). Habitual participation in LTPA was associated with a 23% reduced risk of MetS (OR = 0.77; 95% CI: 0.60 1.03). There was an inverse trend of MetS risk with amount of LTPA (P = 0.016). Compared with non-exercisers, those who exercised <150 min/week had a 21% reduced risk of MetS (AOR = 0.79; 95% CI 0.60-1.04). Individuals who exercised > or = 150 min/week, compared with non-exercisers, had a 42% reduced risk of MetS (AOR = 0.58; 95% CI: 0.36-0.93). Associations of similar magnitudes were observed when men and women were studied separately. CONCLUSION: These data document a high prevalence of MetS and suggest an association with LTPA among urban dwelling Peruvians. Further prospective studies are needed to confirm these observations and to examine interventions that may promote increased physical activity in this population. PMID- 19563446 TI - Choline supplementation reduces oxidative stress in mouse model of allergic airway disease. AB - BACKGROUND: Asthma is a multi-factorial inflammatory disease associated with increased oxidative stress and altered antioxidant defences. We have evaluated the effect of choline on oxidative stress in a mouse model of airway disease. MATERIALS AND METHODS: Balb/c mice were sensitised with 100 microg of ovalbumin on days 0 and 14, and challenged with aerosolized ovalbumin on days 25-27. Mice were administered 1 mg kg(-1) of choline via oral gavage or intranasal route on days 14-27. Mice were also administered 100 mg kg(-1) of alpha-lipoic acid as standard antioxidant. Total cell counts, eosinophils and eosinophil peroxidase (EPO) activity were determined in bronchoalveolar lavage (BAL) fluid. Reactive oxygen species (ROS), lipid peroxidation and isoprostanes levels were measured in BAL fluid. IL-13 and tumour necrosis factor-alpha (TNF-alpha) levels were also measured in BAL fluid and spleen cell culture supernatant. Nuclear factor kappaB (NFkappaB) p65 protein expression was measured after last ovalbumin challenge in nuclear and cytosolic extracts of lungs. RESULTS: Compared with ovalbumin challenged mice, choline and alpha-lipoic acid treated mice had significantly reduced eosinophilic infiltration and EPO activity in BAL fluid. Choline and alpha-lipoic acid treatment reduced ROS production and isoprostanes level significantly in BAL fluid and thus suppressed oxidative stress. Choline and alpha-lipoic acid administration by either route decreased lipid peroxidation levels and down regulated NFkappaB activity. Further, choline and/or alpha-lipoic acid treatment suppressed TNF-alpha level significantly as compared with that of ovalbumin-challenged mice. CONCLUSIONS: Choline administration reduces oxidative stress possibly by modulating the redox status of the cell and inhibits inflammatory response in a mouse model. PMID- 19563447 TI - Fatty acids as metabolic mediators in innate immunity. AB - BACKGROUND: Increasing data support the hypothesis of a local and systemic crosstalk between adipocytes and monocytes mediated by fatty acids. The aim of this study was to characterize the immunomodulatory effects of a large panel of fatty acids on cytokines and chemokines in monocytic THP-1 cells and primary human monocytes. We tested whether anti-inflammatory fatty acids are able to inhibit the binding of lipopolysaccharide (LPS) to its receptor, toll-like receptor/MD-2 (TLR4/MD-2). MATERIALS AND METHODS: Resistin, monocyte chemoattractant protein-1 (MCP-1) and tumour necrosis factor (TNF) were measured by enzyme-linked immunosorbent assay. Proteins were analysed by Western blot. A designed Flag-tagged TLR4/MD-2 fusion protein (LPS trap) was used to investigate the effect of fatty acids on binding of LPS to its receptor. In 30 patients with type 2 diabetes mellitus (T2D), the correlation of serum triglyceride levels with LPS-induced monocyte activation was analysed. RESULTS: Eleven fatty acids investigated exerted differential effects on the monocytic release of cytokines and chemokines. Eicosapentaenoic acid had potent anti-inflammatory effects on human primary monocytes and THP-1 cells; 100 and 200 microM eicosapentaenoic acid dose-dependently inhibited LPS binding to the LPS trap. LPS-induced release of monocytic MCP-1 and TNF was significantly and positively correlated with serum triglyceride levels in 30 patients with T2D. CONCLUSIONS: Monocytic activation is differentially regulated by fatty acids and depends on triglyceride levels in T2D. The main finding of the present study shows that eicosapentaenoic acid inhibits the specific binding of LPS to TLR4/MD-2. Eicosapentaenoic acid represents a new anti-inflammatory LPS-antagonist. PMID- 19563448 TI - Calcitonin and carcinoembryonic antigen doubling times as prognostic factors in medullary thyroid carcinoma: a structured meta-analysis. AB - CONTEXT: In the management of patients with medullary thyroid carcinoma (MTC), calcitonin doubling time (dt) has gained interest as an independent predictor of recurrence and survival. OBJECTIVE: To perform a structured meta-analysis of the diagnostic value of calcitonin dt, carcinoembryonic antigen (CEA) dt and the combination and to define dt strata with the highest predictive power. Design The study was a meta-analysis using individual data. METHODS: Ten studies containing data on the post-operative kinetics of tumour marker(s) and (recurrence free) survival were included. RESULTS: Calcitonin- and CEA-dt are significant indicators for survival (hazard ratios (HR) 21.52 respectively infinite for dt 0 1 year compared to dt >1 year) and recurrence (HR 5.33 respectively 6.80 for dt 0 1 year compared to dt >1 year). The highest predictive power was found for the dt classification 0-1 year vs. >1 year. CEA dt has a higher predictive value than calcitonin dt in the subgroup of patients for which both parameters were available. CONCLUSION: The dts of both calcitonin and CEA are strong prognostic indicators for MTC recurrence and death. CEA dt has a higher predictive value than calcitonin dt and therefore measuring both tumour markers is essential for proper risk stratification. PMID- 19563449 TI - 'Cannibalism' (cell phagocytosis) does not differentiate reactive renal tubular cells from urothelial carcinoma cells. AB - OBJECTIVE: Cannibalism of one cell by another in voided urine cytology has been considered a cytological feature for differentiating urothelial carcinoma (UC) from benign lesions. Recently, however, we observed cannibalism in voided urine obtained from patients with renal glomerular disease (RGD). The purpose of this study was to determine the cytomorphological and immunocytochemical characteristics of cannibalism in voided urine from RGD. METHODS: Seventy cytology specimens of voided urine were examined and the findings were compared with the histological findings. In addition, we compared the cytomorphological and immunocytochemical differences in cannibalism found in RGD and cases of UC selected as showing cannabilism. RESULTS: Cannibalism in voided urine was found in three (5.5%) of 55 RGD cases. The finding was measured as (1+) < 5 cells, (2+) 5-20 cells, and (3+) > 20 cells and was (1+) in all three RGD cases, compared with 6.7%, 60% and 33.3% respectively in 15 UC cases. Differences in low cellularity cases (1+) and moderate to high cellularity cases (2+ or 3+) were statistically significant between RGD (3 and 0) and UC (1 and 14) (P=0.005). The maximum diameter of cannibalized cells in RGD was 24.3-33.0 microm (mean 29.8 microm) versus 18.0-30.4 microm (mean 23.3 microm) in UC (P=0.004). Necrosis and isomorphic erythrocytes were absent in RGD, but were found in 46.7% and 86.7%, respectively, of UC cases (P=0.245 and P=0.012). Dysmorphic erythrocytes were identified in all three cases with RGD and 13.3% of UC (P=0.012). Vimentin reactivity was found in all cases with cannibalism in RGD, but never in UC (P=0.001). CONCLUSIONS: Our results demonstrated that cannibalism in voided urine is present not only in UC but also in RGD. Furthermore, we showed that cellularity of cannibalism, vimentin reactivity and background differed significantly and can be used for differential diagnosis between the two groups. PMID- 19563450 TI - Thromboembolic events associated with Neuroform stent in endovascular treatment of intracranial aneurysms. AB - BACKGROUND: The incidence of thromboembolic events associated with Neuroform stent (Boston Scientific Target, Fremont, CA) is known from previous studies but there are uncertainties of scale. PURPOSE: To report our rate of ischemic events associated with Neuroform stent. METHODS: Consecutive patients treated with Neuroform stent for intracranial aneurysms were prospectively enrolled from January 2003 to August 2006. Thromboembolic events as well as clinical outcomes were measured. Mean follow-up was 12 months. RESULTS: Successful stent deployment was achieved in (65/67) 97% of patients without any ischemic event. However, postoperative thromboembolic events were observed in 3 patients despite being on clopidogrel and aspirin. These 3 patients demonstrated poor platelet inhibitions in platelet aggregation (aggregometry) studies, and were successfully treated with intravenous eptifibatide with good outcome. The majority of the patients had good outcomes [Glasgow Outcome Score (GOS) 5 or National Institute of Health Stroke Scale (NIHSS) 0 in (63/67) 94%, GOS 4 or NIHSS 2 in 1 patient, and GOS 3 or NIHSS 4 was observed in 3 cases]. CONCLUSION: Our study reveals that the thromboembolic events associated with Neuroform stent may present in a delayed fashion. These events can be successfully treated with good outcome. Therefore, postoperative close follow-up is strongly recommended for all Neuroform stent treated patients. PMID- 19563451 TI - Continua of specificity and virulence in plant host-pathogen interactions: causes and consequences. AB - Ecological, evolutionary and molecular models of interactions between plant hosts and microbial pathogens are largely based around a concept of tightly coupled interactions between species pairs. However, highly pathogenic and obligate associations between host and pathogen species represent only a fraction of the diversity encountered in natural and managed systems. Instead, many pathogens can infect a wide range of hosts, and most hosts are exposed to more than one pathogen species, often simultaneously. Furthermore, outcomes of pathogen infection vary widely because host plants vary in resistance and tolerance to infection, while pathogens are also variable in their ability to grow on or within hosts. Environmental heterogeneity further increases the potential for variation in plant host-pathogen interactions by influencing the degree and fitness consequences of infection. Here, we describe these continua of specificity and virulence inherent within plant host-pathogen interactions. Using this framework, we describe and contrast the genetic and environmental mechanisms that underlie this variation, outline consequences for epidemiology and community structure, explore likely ecological and evolutionary drivers, and highlight several key areas for future research. PMID- 19563452 TI - Pollen resistance to water in 80 angiosperm species: flower structures protect rain-susceptible pollen. AB - Flowers exhibit adaptive responses to biotic and abiotic factors. It remains unclear whether pollen susceptibility to rain damage plays a role in the evolution of floral form. We investigated flower performance in rain and compared pollen longevity in dry conditions, pure water and solutions with different sucrose concentrations in 80 flowering species from 46 families with diverse floral shapes and pollination modes. A pollen viability test showed that pollen longevity in all studied species was greatly reduced by wetting. We found that pollen of species with complete protection by flower structures was susceptible to water damage and a high proportion of resistant pollen occurred in unprotected species. Flowers whose structures expose pollen to rain may also reduce rain damage through temporal patterns of pollen presentation. This prediction was supported by our direct measurement of pollen presentation duration on rainy days. Our observations showed that variation in pollen performance in water was associated with differences in floral forms. Water-resistant pollen and extended pollen presentation duration were favored by selection via rain contact in species in which pollen was not protected from rain. These findings support the functional hypothesis that flower structures protect susceptible pollen from rain, demonstrating that rain acts as a force shaping floral form. PMID- 19563453 TI - A review of the effect of the psychosocial working environment on physiological changes in blood and urine. AB - The aim of the present survey was to provide a literary review of current knowledge of the possible association between the psychosocial working environment and relevant physiological parameters measured in blood and urine. Literature databases (PubMed, Toxline, Biosis and Embase) were screened using the key words job, work-related and stress in combination with selected physiological parameters. In total, 51 work place studies investigated the associations between the psychosocial working environment and physiological changes, of which 20 were longitudinal studies and 12 population-based studies. The studied exposures in work place/population-based studies included: job demands (26/8 studies), job control (24/10 studies), social support and/or leadership behaviour (12/3 studies), effort-reward imbalance (three/one studies), occupational changes (four studies), shift work (eight studies), traumatic events (one study) and other (five studies). The physiological responses were catecholamines (adrenaline, noradrenaline) (14 studies), cortisol (28 studies), cholesterol (23 studies), glycated haemoglobinA(1c) (six studies), testosterone (nine studies), oestrogens (three studies), dehydroepiandrosterone (six studies), prolactin (14 studies), melatonin (one study), thyroxin (one study), immunoglobulin (Ig) A (five studies), IgG (four studies), IgM (one study) and fibrinogen (eight studies). In general, fibrinogen and catabolic indicators, defined as energy releasing, were increased, whereas the anabolic indicators defined as constructive building up energy resources were decreased when the psychosocial working environment was perceived as poor. In conclusion, in this review the association between an adverse psychosocial working environment and HbA(1c), testosterone and fibrinogen in serum was found to be a robust and potential candidate for a physiological effect of the psychosocial working environment. Further, urinary catecholamines appear to reflect the effects of shift work and monotonous work. PMID- 19563454 TI - Carbohydrate intake, serum lipids and apolipoprotein E phenotype show association in children. AB - AIM: To study the association between carbohydrate intake and serum lipids in children, and influence of apolipoprotein E phenotype (apoE) on the association. SUBJECTS/METHODS: A total of 644 children from a prospective, randomized atherosclerosis prevention trial (STRIP) participated in this longitudinal study at age 5 (n = 644), 7 (n = 585) and 9 (n = 550) years. ApoE phenotype, fasting triglyceride, total, low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol concentrations and 4-day food records were analysed. RESULTS: An increase in the total carbohydrate intake by 1 E% (percentage of total daily energy intake) associated with a decrease in HDL cholesterol by 0.006 mmol/L (p < 0.001) when adjusted for saturated, monounsaturated and polyunsaturated fatty acid, age, gender, body mass index and STRIP study group. The inverse association between total carbohydrate intake and HDL cholesterol was evident in children with apoE3 (p < 0.001) or apoE4 (p < 0.001), but not in those with apoE2 (p = 0.78). An increase in total carbohydrate intake by 1 E% increased triglycerides by 0.02 mmol/L (p < 0.001) independently of apoE phenotype, while 1 E% increase in sucrose intake increased triglycerides by 0.01 mmol/L (p < 0.001). CONCLUSION: Carbohydrate intake has a relatively small effect on serum lipids in children. Children with the apoE3 or E4 but not with E2 phenotype show reduction in HDL cholesterol with increasing carbohydrate intake indicating that genetic and environmental factors interact with children's lipoprotein metabolism. PMID- 19563455 TI - Baby walkers: a perspective from Turkey. AB - AIM: To determine in an urban population in Turkey, the frequency of baby walker (BW) use, beliefs and attitudes of mothers regarding BWs, frequency of BW-related injuries and whether families receive counselling from their paediatricians about BWs or not. METHODS: Children aged 2 months to 5 years who attended the well child care clinics of Fatih University Hospital in Ankara comprised the sample. A semi-structured questionnaire was verbally administered. RESULTS: Of 495 children, 75.4% was found to use BW. Female gender [odds ratio (OR) = 1.82, 95% confidence interval (CI) =1.19-2.78) and lower maternal education (OR = 0.37, 95% CI = 1.18-0.74) were found to be associated with BW use. Frequencies of injuries associated with BWs were low (7.8%). Only 92 (18.6%) families received appropriate counselling by their paediatricians. CONCLUSION: The results of our study show that BW use is common in urban Turkey. Families are not informed about the lack of benefits and potential hazards of BWs and base reasons of using and not using on unique cultural beliefs rather than informed, evidence-based decisions. Paediatricians in Turkey and potentially other countries require information about the need for counselling families about the hazards associated with BW use. PMID- 19563456 TI - Natriuretic peptides: linking heart and adipose tissue in obesity and related conditions--a systematic review. AB - The objective of this study was to investigate the association between natriuretic peptides, obesity and related comorbidities. A systematic review of the English language literature from 1996 to 2008 was performed with Pubmed/MEDLINE and the ISI Web of Knowledge. 'Natriuretic peptides', 'atrial natriuretic factor', 'brain natriuretic peptide', 'obesity', 'body mass index', 'lipolysis' and 'adipose tissue' were used as Mesh terms. We also conducted a handle search among the references of the original articles selected. Finally, seventy-five studies were considered eligible for inclusion in the review. Natriuretic peptides are widely known as body homeostasis regulators. Recently, their action as lipolytic agents has been identified. Obese patients, especially those with hypertension and metabolic risk factors, have reduced plasma levels of natriuretic peptides. Whether this precedes or follows obesity and its complications remains undefined. The lipolytic effect of natriuretic peptides indicates that they may be involved in the pathophysiology of obesity. In general, studies with obese patients support paradoxical reduced levels of natriuretic peptides. However, the selection of subjects and classification of obesity and heart failure varied among the reviewed studies, rendering comparison unreliable. PMID- 19563457 TI - Effects of adjustable gastric banding on gastroesophageal reflux and esophageal motility: a systematic review. AB - Controversial opinions exist concerning the effect of laparoscopic adjustable gastric banding on gastroesophageal reflux. MEDLINE and EMBASE databases were searched for relevant studies on patients undergoing adjustable gastric banding. Data are expressed in mean (range). Twenty studies were identified with a total of 3307 patients. The prevalence of reflux symptoms decreased postoperatively from 32.9% (16-57) to 7.7% (0-26.9) and medication use from 27.5% (16-38.5) to 9.5% (3.1-19.2). Newly developed reflux symptoms were found in 15% (6.1-20) of the patients. The percentage of esophagitis decreased postoperatively from 33.3% (19.4-61.6) to 27% (2.3-60.8). Newly developed esophagitis was observed in 22.9% (0-38.4). Pathological reflux was found in 55.8% (34.9-77.4) preoperatively and postoperatively in 29.4% (0-41.7) of the patients. Lower esophageal sphincter pressures increased from 12.9 to 16.9 mmHg (11.3-21.4). Lower esophageal sphincter relaxation decreased from 100% to 79.7% (58-86). The percentage of dysmotility increased from 3.5% (0-10) to 12.6% (0-25). Adjustable gastric banding has anti-reflux properties resulting in resolution or improvement of reflux symptoms, normalized pH monitoring results and a decrease of esophagitis on short term. However, worsening or newly developed reflux symptoms and esophagitis are found in a subset of patients during longer follow-up. PMID- 19563459 TI - Primary antimicrobial resistance of Helicobacter pylori in children during the past 9 years. AB - BACKGROUND: Antimicrobial resistance of Helicobacter pylori is a growing problem in clinical practice, particularly clarithromycin resistance. The aim of the present study was therefore to investigate the prevalence of H. pylori resistance to antimicrobial agents in Japanese children. METHODS: A total of 61 H. pylori strains isolated from children (mean age, 12.6 years; range, 4-18 years) between 1999 and 2007 were studied for primary antimicrobial resistance, using a microdilution method. In addition, the eradication rate with lansoprazole-based triple regimens was determined. RESULTS: The overall resistance rate of clarithromycin, amoxicillin and metronidazole was 36.1%, 0% and 14.8%, respectively. Resistance to both clarithromycin and metronidazole was detected in 6.6% of the strains. The rate of clarithromycin-resistant strains was 32.4% from 1999 to 2002 and 40.7% from 2003 to 2007, and clarithromycin minimum inhibitory concentration at which 90% of the isolates were inhibited (MIC(90)) increased fourfold from 1999-2002 to 2003-2007, with all clarithromycin-resistant strains showing low-level resistance. Metronidazole resistance rates were not different between these two study periods. Regimens involving amoxicillin and clarithromycin (n= 49) had a higher eradication rate in clarithromycin susceptible strains (97.1%) than in the resistant strains (57.1%; P < 0.001). There was no difference in the eradication rate between 7 day and 10 or 14 day courses of the regimens (P= 0.53). The regimen with amoxicillin and metronidazole produced successful eradication in all nine patients with clarithromycin resistant strains. CONCLUSIONS: Clarithromycin resistance of H. pylori is high, and triple regimen treatment containing clarithromycin should be decided based on susceptibility to the agent. PMID- 19563458 TI - Novel SCN1A mutations in Indonesian patients with severe myoclonic epilepsy in infancy. AB - BACKGROUND: Severe myoclonic epilepsy in infancy (SMEI) and borderline SMEI (SMEB) are caused by a mutation in SCN1A, which encodes a voltage-gated sodium channel alpha1-subunit protein. Although many mutations in SCN1A have been associated with clinical features of SMEI or SMEB from different ethnic groups, there have been no such reports from the South-East Asian populations so far. METHODS: Patients 1 and 2 were Indonesian children diagnosed as having SMEI and SMEB based on their clinical features. SCN1A was screened for mutations using a combination of polymerase chain reaction and denaturing high-performance liquid chromatography. Nucleotide substitutions were confirmed on direct sequencing. RESULTS: In patient 1, a G-to-A heterozygous transition was detected at nucleotide 4834 (c.4834G>A) in exon 25, leading to substitution of valine with isoleucine at amino acid position 1612 (p.V1612I) in the SCN1A protein. In patient 2 a T-to-G heterozygous transversion was identified at nucleotide 5266 (c.5266T>G) in exon 26, leading to substitution of cysteine with glycine at amino acid 1756 (p.C1756G) in the SCN1A protein. Both amino acid substitutions might disrupt these highly conserved regions in species from drosophila to human, leading to dysfunction of the protein. p.V1612I and p.C1756G were determined as disease-causing mutations due to their absence in the control population. CONCLUSION: The first cases of SMEI and SMEB are reported in South-East Asian populations. Two novel SCN1A mutations are also identified in these patients, p.V1612I and p.C1756G, which may lead to neuronal excitability or convulsions. PMID- 19563460 TI - Grade III lipaemia retinalis in a newborn. PMID- 19563461 TI - Galactofuranose attenuates cellular adhesion of Aspergillus fumigatus. AB - Galactofuranose (Galf) is a major molecule found in cell wall polysaccharides, secreted glycoproteins, membrane lipophosphoglycans and sphingolipids of Aspergillus fumigatus. The initial step in the Galf synthetic pathway is the re arrangement of UDP-galactopyranose to UDP-Galf through the action of UDP galactopyranose mutase. A mutant lacking the AfUGM1 gene encoding the UDP galactopyranose mutase has been constructed. In the mutant, though there is a moderate reduction in the mycelial growth associated with an increased branching, it remains as pathogenic and as resistant to cell wall inhibitors and phagocytes as the wild-type parental strain. The major phenotype seen is a modification of the cell wall surface that results in an increase in adhesion of the mutants to different inert surfaces (glass and plastic) and epithelial respiratory cells. The adhesive phenotype is due to the unmasking of the mannan consecutive to the removal of galactofuran by the ugm1 mutation. Removal of the mannan layer from the mutant surface by a mannosidase treatment abolishes mycelial adhesion to surfaces. PMID- 19563463 TI - Pharmacophore identification and validation study of CK2 inhibitors using CoMFA/CoMSIA. AB - Protein kinase CK2, also known as casein kinase-2, has been found to be involved in cell growth, proliferation and suppression of apoptosis, which is related to human cancers. The series of compounds were identified as casein kinase-2 inhibitors and their inhibitory activities are a function of a variation of their structures. The current study deals with the pharmacophore identification and, accordingly, the three-dimensional quantitative structure-activity relationship model development using Pharmacophore Alignment and Scoring Engine. Several hypotheses were developed for the molecular alignments. On the basis of statistical values, the best-fitted model was identified and the same alignment was used for 3D-QSAR using comparative molecular field analysis/comparative molecular similarity index analysis. Both the CoMFA (R(2)(CV) = 0.58, R(2) = 0.82 and r(2)(pred) = 0.62) and the comparative molecular similarity index analysis (R(2)(CV) = 0.74, R(2) = 0.98 and r(2)(pred) = 0.81) gave reasonable results. Besides pharmacophore-based alignment, the maximum common substructure-based alignment was also used for the comparative molecular field analysis and comparative molecular similarity index analysis. The pharmacophore-based alignment was more prominent and it has provided important information for the modelling of potent inhibitors. The overall study implies that a highly positive and bulky group with H-bond donating property is desirable around the nitrogen atom adjacent to the pyrrolidine ring. PMID- 19563462 TI - The perplexing functions and surprising origins of Legionella pneumophila type IV secretion effectors. AB - Only a limited number of bacterial pathogens evade destruction by phagocytic cells such as macrophages. Legionella pneumophila is a Gram-negative gamma proteobacterial species that can infect and replicate in alveolar macrophages, causing Legionnaires' disease, a severe pneumonia. L. pneumophila uses a complex secretion system to inject host cells with effector proteins capable of disrupting or altering the host cell processes. The L. pneumophila effectors target multiple processes but are essentially aimed at modifying the properties of the L. pneumophila phagosome by altering vesicular trafficking, gradually creating a specialized vacuole in which the bacteria replicate robustly. In nature, L. pneumophila is thought to parasitize free-living protists, which may have selected for traits that promote virulence of L. pneumophila in humans. Indeed, many effector genes encode proteins with eukaryotic domains and are likely to be of protozoan origin. Sustained horizontal gene transfer events within the protozoan niche may have allowed L. pneumophila to become a professional parasite of phagocytes, simultaneously giving rise to its ability to infect macrophages, cells that constitute the first line of cellular defence against bacterial infections. PMID- 19563464 TI - Acute allergic skin response as a new tool to evaluate the allergenicity of whey hydrolysates in a mouse model of orally induced cow's milk allergy. AB - Hypoallergenic milk formulae are used for cow's milk allergic infants and may be a good option for infants at risk. Clinical studies have shown that the protein source or the hydrolysis methodology used may influence the effectiveness in infants stressing the importance of adequate pre-clinical testing of hypoallergenic formulae in an in vivo model of orally induced cow's milk allergy. This study was undertaken to introduce a new read-out system to measure the residual allergenicity of whey hydrolysates on both the sensitization and challenge phase of orally induced cow's milk allergy in mice. Mice were sensitized orally to whey or a partial whey hydrolysate (pWH) to measure the residual sensitizing capacity. To predict the residual allergenicity of hydrolysates, whey allergic mice were challenged in the ear with pWH, extensive whey hydrolysate or an amino acid-based formula. An acute allergic skin response (ear swelling at 1 h), whey-specific serum antibodies, and local MCP-1 concentrations were measured. In contrast to whey, oral sensitization with pWH did not result in the induction of whey-specific antibodies, although a minor residual skin response to whey was observed after challenge. Skin exposure to whey hydrolysates showed a hydrolysation dependent reduction of the acute allergic skin response in whey allergic mice. In contrast to whey, skin exposure to pWH did not enhance tissue MCP-1 levels. The acute allergic skin response in mice orally sensitized to cow's milk proteins reveals a new pre-clinical tool which might provide information about the residual sensitizing capacity of hydrolysates supporting the discussion on the use of hypoallergenic formulae in high risk children. This mouse model might be a relevant model for the screening of new hypoallergenic formulae aimed to prevent or treat cow's milk allergy. PMID- 19563465 TI - Feasibility of a new method to collect exhaled breath condensate in pre-school children. AB - Exhaled breath condensate (EBC) is a promising non-invasive method to assess respiratory inflammation in adults and children with lung disease. Especially in pre-school children, condensate collection is hampered by long sampling times because of open-ended collection systems. We aimed to assess the feasibility of condensate collection in pre-school children using a closed glass condenser with breath recirculation system, which also collects the residual non-condensed exhaled breath, and subsequently recirculates it back into the condenser. Condensate was collected before and after breath recirculation in 70 non-sedated pre-school children with and without recurrent wheeze. Cytokines (IL-4, IL-5, IL 6, IL-8, IL-10, IL-12p70, IL-13, TNF-alpha) were measured in 50 MUl samples using ultrasensitive multiplexed liquid bead array. The success rate of condensate collection increased from 64% (without recirculation) to 83% (after breath recirculation), and mean condensate volume from 214 to 465 MUl respectively. Detection of cytokines was successful in 95-100% of samples. Cytokine concentrations before and after breath recirculation were not different (p > 0.232). In asthmatic children, only TNF-alpha concentrations were significantly decreased, compared to non-asthmatics. In pre-school children, the collection of EBC is feasible using a new closed glass condenser with breath recirculation system. This new method may help to assess - non-invasively - cytokine profiles in asthmatic and non-asthmatic pre-school children. PMID- 19563466 TI - Efficacy of tacrolimus 0.03% ointment as second-line treatment for children with moderate-to-severe atopic dermatitis: evidence from a randomized, double-blind non-inferiority trial vs. fluticasone 0.005% ointment. AB - Tacrolimus 0.03% ointment is licensed for second-line treatment of children with atopic dermatitis (AD). Although data are available from clinical trials, no study has enrolled only second-line patients. This double-blind, non-inferiority study compared tacrolimus 0.03% and fluticasone 0.005% ointments in children with moderate-to-severe AD, who had responded insufficiently to conventional therapies. Children (aged 2-15 yr) were randomized to tacrolimus ointment (n = 240) or fluticasone ointment (n = 239), twice daily until clearance or for a maximum of 3 wk and, if lesions remained, once daily for up to 3 wk further. Primary end-point was week 3 response rate (improvement of >or=60% in modified Eczema Area and Severity Index and not withdrawn for lack of efficacy). Secondary end-points included pruritus and sleep quality, global assessment of clinical response, incidence of new flares and safety. Response rates were 86.3% with tacrolimus ointment and 91.5% with fluticasone. Lower limit of the 95% confidence interval was -11.8%, exceeding the non-inferiority limit of -15% and meeting the primary end-point. Moderate or better improvement on the physicians' global assessment occurred in 93.6% and 92.4% of patients in the tacrolimus ointment and fluticasone arms, respectively, while median pruritus scores improved by 84.0% and 91.5%. Sleep quality improved by approximately 92% in both treatment arms. After day 21, new flare-up occurred in 5.5% and 11.3% of patients receiving tacrolimus ointment and fluticasone, respectively; mean times to new flares were 6.5 +/- 5.0 and 8.6 +/- 5.2 days. Adverse events were similar between the two arms, with the exception of application-site skin burning sensation in the tacrolimus ointment group. In conclusion, efficacy of tacrolimus 0.03% ointment as second-line treatment was not inferior to that of fluticasone 0.005% ointment, with similar benefits on global disease improvement and quality of sleep. PMID- 19563467 TI - Serum hepcidin concentration in chronic haemodialysis patients: associations and effects of dialysis, iron and erythropoietin therapy. AB - BACKGROUND: Hepcidin, a liver-derived peptide induced by iron overload and inflammation, is a major regulator of iron homeostasis. As hepcidin decreases gastrointestinal iron absorption and recirculation from monocytes, over expression is associated with the development of anaemia. METHODS: We studied the associations between circulating hepcidin levels and various laboratory parameters related to anaemia and/or inflammation in 20 patients on chronic haemodialysis. Furthermore, we determined the impact of dialysis and iron and/or erythropoietin (rhEpo) supplementation therapy on hepcidin serum concentrations. The patients were withheld from iron and rhEpo for 2 weeks before study entry. Hepcidin was measured by liquid chromatography-mass spectrometry (LC-MS/MS); serum iron and haematological parameters, cytokines and pro-hepcidin by commercially available enzyme-linked immunosorbent assays (ELISA) or standard automated methods. RESULTS: While hepcidin levels at baseline were not correlated to pro-hepcidin, interleukin-6 or transforming growth factor-beta concentrations, we found significant associations with reticulocyte count (r = -0.55; P = 0.015), serum iron (r = 0.7; P = 0.004) and ferritin levels (r = 0.63; P = 0.004) and transferrin saturation (r = 0.69, P = 0.001). Dialysis using either a high or a low flux biocompatible dialyser resulted in a significant decrease of hepcidin concentrations, which returned to pre-dialysis values before the next dialysis session. When studying the effects of anaemia treatment, we observed a significant reduction of hepcidin levels following administration of rhEpo but not iron. CONCLUSIONS: Hepcidin levels in stable haemodialysis patients appear to reflect systemic iron load, but not inflammation. Due to the negative association between reticulocyte counts and hepcidin, the reduction of circulating hepcidin concentrations by dialysis and/or rhEpo treatment may positively affect erythropoiesis. PMID- 19563468 TI - Methods to assess impaired post-prandial metabolism and the impact for early detection of cardiovascular disease risk. AB - Post-prandial lipaemia has emerged as a key contributor to cardiovascular disease (CVD) risk and progression. Specifically, delayed clearance of chylomicrons (CM) and their remnants increase the delivery of triglyceride and cholesteryl ester to the vessel wall and can accelerate the progression of atherosclerosis, which may be particularly pertinent to individuals with insulin resistance and/or obesity. As the number of studies linking post-prandial metabolism and chronic disease increases, interest has grown in the use of parameters reflecting CM metabolism as a possible indicator of early CVD risk. This, in turn has raised the question of what method might be most appropriate to detect CM and their remnants in plasma accurately. However, the handful of techniques able to measure CM metabolism (triglyceride-rich lipoprotein fractions; remnant-lipoprotein cholesterol; retinyl esters, CM-like emulsion; sodium dodecyl sulphate polyacrylamide gel electrophoresis; immunoblotting, enzyme-linked immunoabsorbent assays; C(13) breath test; capillary finger prick) differ in their specificity, cost and applicability in research and in the clinical setting. In this review, we explore the scientific and clinical implications of CM methodology to better understand early risk assessment of CVD. We raise ongoing issues of the need to appreciate differential separation of very low-density lipoprotein and CM fractions, as well as to identify the technical basis for imprecision between assays for apolipoprotein B48. We also highlight emerging issues with respect to the practicality of measuring post-prandial metabolism in large clinical studies and offer opinions on the appropriateness of existing techniques in this field. PMID- 19563469 TI - Habitat and roe deer fawn vulnerability to red fox predation. AB - 1. Notwithstanding the growing amount of literature emphasizing the link between habitat, life-history traits and behaviour, few empirical studies investigated the combined effect of these parameters on individual predation risk. We investigated direct and indirect consequences of habitat composition at multiple spatial scales on predation risk by red foxes on 151 radio-monitored roe deer fawns. We hypothesized that the higher resource availability in fragmented agricultural areas increased predation risk because of: (i) shorter prey movements, which may increase predictability; (ii) larger litter size and faster growth rates, which may increase detectability in species adopting a hiding neonatal anti-predator strategy. The sharing of risky habitat among littermates was expected to promote whole-litter losses as a result of predation. 2. The landscape-scale availability of agricultural areas negatively affected pre weaning movements, but did not influence growth rates or litter size. Predation risk was best described by the interplay between movements and fine-scale habitat fragmentation: a higher mobility increased the encounter rate and predation risk in highly fragmented home ranges, while it reduced predation risk in forest dominated areas with clumped resources because of decreased predictability. This is one of the first demonstrations that movement patterns can be an efficient anti-predator strategy when adjusted to local conditions. 3. In accordance with previous studies documenting the existence of family effects (i.e. non independence among siblings) in survival, littermates survived or died together more often than expected by chance. In addition, our study specifically demonstrated the occurrence of behaviourally mediated family effects in predation risk: after a fox killed one fawn the probability of a sibling being killed within a few days rose from 20% to 47%, likely because of the win-stay strategy (i.e. return to a previously rewarding site) adopted by the predator. Hence, the predator's hunting strategy has the potential to raise fawn mortality disproportionately to predator abundance. 4. There is increasing evidence that fawns inhabiting highly productive predator-free habitats are granted lifetime fitness benefits; these potential advantages, however, can be cancelled out when predation risk increases in the very same high-productivity areas, which might thus turn into attractive sinks. PMID- 19563470 TI - Classifying movement behaviour in relation to environmental conditions using hidden Markov models. AB - 1. Linking the movement and behaviour of animals to their environment is a central problem in ecology. Through the use of electronic tagging and tracking (ETT), collection of in situ data from free-roaming animals is now commonplace, yet statistical approaches enabling direct relation of movement observations to environmental conditions are still in development. 2. In this study, we examine the hidden Markov model (HMM) for behavioural analysis of tracking data. HMMs allow for prediction of latent behavioural states while directly accounting for the serial dependence prevalent in ETT data. Updating the probability of behavioural switches with tag or remote-sensing data provides a statistical method that links environmental data to behaviour in a direct and integrated manner. 3. It is important to assess the reliability of state categorization over the range of time-series lengths typically collected from field instruments and when movement behaviours are similar between movement states. Simulation with varying lengths of times series data and contrast between average movements within each state was used to test the HMMs ability to estimate movement parameters. 4. To demonstrate the methods in a realistic setting, the HMMs were used to categorize resident and migratory phases and the relationship between movement behaviour and ocean temperature using electronic tagging data from southern bluefin tuna (Thunnus maccoyii). Diagnostic tools to evaluate the suitability of different models and inferential methods for investigating differences in behaviour between individuals are also demonstrated. PMID- 19563471 TI - Organogenesis of the juxta-oral organ in mice. AB - The juxta-oral organ is a bilateral organ in the mammalian bucca. It consists of epithelial cords with surrounding mesenchyme. It develops from embryonic oral epithelium, but its macroscopic morphology in mice is less studied and seems to be very different from that of humans. The juxta-oral organ in mice extends more widely from the subcutaneous tissue of the mandible near the lateral fascia of the masseter to the submucosa of the soft palate. In this paper, we report that the mutant mouse allele Bmp7(lacZ) presented intense lacZ expression in the epithelial component of the juxta-oral organ in its homo- and heterozygous states. The main aims of this study were to show that this mutant mouse allele is suitable for observing macroscopic structure of the juxta-oral organ and to describe the development of this organ during embryonic and postnatal stages. Whole-mount beta-gal staining of this strain of mouse showed that the juxta-oral organ in mice appeared at E12.0 from oral epithelium and lost connection with it before E12.5. Then, the juxta-oral organ extended anteriorly to the lateral fascia of the masseter and posteriorly to the submucosal layer of the soft palate via the orbit. The mature juxta-oral organ had no connection to other epithelia such as those of the bucca and parotid duct. It persisted until adulthood and there seemed to be no tendency to regress. Transmission electron microscopy showed that each part of the juxta-oral organ was an epithelial cord surrounded by a basement membrane and mesenchymal tissue. PMID- 19563472 TI - Identification of a stem cell niche in the zone of Ranvier within the knee joint. AB - A superficial lesion of the articular cartilage does not spontaneously self repair and has been suggested to be partly due to lack of progenitor cells within the joint that can reach the site of injury. To study whether progenitor cells are present within the joint, 3-month-old New Zealand white rabbits were exposed to bromodeoxyuridine (BrdU) for 12 consecutive days and were then sacrificed 4, 6, 10, 14, 28 and 56 days after the first BrdU administration. Presence of BrdU and localization of progenitor markers were detected using immunohistochemistry. After 10 days of BrdU exposure, BrdU-positive cells, i.e. proliferating cells, were abundantly detected in the epiphyseal plate, the perichondrial groove of Ranvier, and in all zones of the articular cartilage. After a wash-out period, BrdU-positive cells were still present, i.e. those considered to be progenitor cells, in these regions of the knee except for the proliferative zone of the epiphyseal plate. Cells in the perichondrial groove of Ranvier were further positive for several markers associated with progenitor cells and stem cell niches, including Stro-1, Jagged1, and BMPr1a. Our results demonstrate that a small population of progenitor cells is present in the perichondrial groove of Ranvier as well as within the articular cartilage in the knee. The perichondrial groove of Ranvier also demonstrates the properties of a stem cell niche. PMID- 19563474 TI - Application of array-based whole genome scanning technologies as a cytogenetic tool in haematological malignancies. AB - Karyotypic analysis provides useful diagnostic information in many haematological malignancies. However, standard metaphase cytogenetics has technical limitations that result in the underestimation of the degree of chromosomal changes. Array based technologies can be used for karyotyping and can supplant some of the shortcomings of metaphase cytogenetics, and include single nucleotide polymorphism arrays (SNP-A) and comparative genomic hybridization arrays (CGH-A). Array-based cytogenetic tools do not rely on cell division, have superb resolution for unbalanced lesions and allow for the detection of copy number neutral loss of heterozygosity, a type of lesion not seen with metaphase cytogenetics. Moreover, genomic array analysis is automated and results can be objectively and systematically analysed using biostatistical algorithms. As a potential advantage over genomic approaches, metaphase cytogenetics can detect balanced chromosomal defects and resolves clonal mosaicism. Initial studies performed in various haematological malignancies indicate the potential of SNP-A based karyotyping as a useful clinical cytogenetic detection tool. The current effort is aimed at developing rational diagnostic algorithms for the detection of somatic defects and the establishment of clinical correlations for novel SNP-A detected chromosomal defects, including acquired somatic uniparental disomy. SNP A can complement metaphase karyotyping and will probably play an important role in clinical cytogenetic diagnostics. PMID- 19563473 TI - Facial musculature in the rhesus macaque (Macaca mulatta): evolutionary and functional contexts with comparisons to chimpanzees and humans. AB - Facial expression is a common mode of visual communication in mammals but especially so in primates. Rhesus macaques (Macaca mulatta) have a well documented facial expression repertoire that is controlled by the facial/mimetic musculature as in all mammals. However, little is known about the musculature itself and how it compares with those of other primates. Here we present a detailed description of the facial musculature in rhesus macaques in behavioral, evolutionary and comparative contexts. Formalin-fixed faces from six adult male specimens were dissected using a novel technique. The morphology, attachments, three-dimensional relationships and variability of muscles were noted and compared with chimpanzees (Pan troglodytes) and with humans. The results showed that there was a greater number of facial muscles in rhesus macaques than previously described (24 muscles), including variably present (and previously unmentioned) zygomaticus minor, levator labii superioris alaeque nasi, depressor septi, anterior auricularis, inferior auricularis and depressor supercilii muscles. The facial muscles of the rhesus macaque were very similar to those in chimpanzees and humans but M. mulatta did not possess a risorius muscle. These results support previous studies that describe a highly graded and intricate facial expression repertoire in rhesus macaques. Furthermore, these results indicate that phylogenetic position is not the primary factor governing the structure of primate facial musculature and that other factors such as social behavior are probably more important. The results from the present study may provide valuable input to both biomedical studies that use rhesus macaques as a model for human disease and disorder that includes assessment of facial movement and studies into the evolution of primate societies and communication. PMID- 19563475 TI - Endocannabinoids render exploratory behaviour largely independent of the test aversiveness: role of glutamatergic transmission. AB - To investigate the impact of averseness, controllability and familiarity of a test situation on the involvement of the endocannabinoid system in the regulation of exploratory behaviour, we tested conventional and conditional cannabinoid receptor type 1 (CB1)-deficient mice in behavioural paradigms with different emotional load, which depended on the strength of illumination and the ability of the animals to avoid the light stimulus. Complete CB1 null-mutant mice (Total-CB1 KO) showed an anxiogenic-like phenotype under circumstances where they were able to avoid the bright light such as the elevated plus-maze and the light/dark avoidance task. Conditional mutant mice lacking CB1 expression specifically in cortical glutamatergic neurons (Glu-CB1-KO), in contrast, failed to show a similar phenotype under the same experimental conditions. However, both mutant lines showed increased avoidance of open arm exploration during a second exposure to the elevated plus-maze. If tested in situations where the fear eliciting light could not be avoided, Total-CB1-KO mice showed increased thigmotaxis in an open field, decreased social investigation and decreased novel object exploration under aversive light conditions, but not under non-aversive low light. This time, Glu-CB1-KO also showed decreased exploratory behaviour towards objects and conspecific juveniles and increased thigmotaxis in the open field. Taking into consideration that the behavioural performance of wild-type mice was only marginally affected by changes in light intensities, these data indicate that the endocannabinoid system renders exploratory behaviour largely independent of the test averseness. This process differentially involves endocannabinoid-controlled glutamatergic transmission, depending on the controllability of the test situation. PMID- 19563476 TI - Therapeutic itineraries of patients with ulcerated forms of Mycobacterium ulcerans (Buruli ulcer) disease in a rural health zone in the Democratic Republic of Congo. AB - OBJECTIVE: To describe lay perceptions of the ulcerated forms of Mycobacterium ulcerans, commonly called Buruli ulcer (BU), and therapeutic itineraries of BU patients in a rural area of the Democratic Republic of Congo. METHODS: Qualitative research consisting of semi-structured interviews of 19 patients with clinical signs of BU and 12 in-depth interviews of confirmed cases allowing for a detailed reconstruction of the itineraries followed. RESULTS: The first symptoms of BU are perceived as mild. The perceived seriousness of the disease increases as the ulceration persists, increases in size or results in complications. Knowledge about the biomedical aetiology of the disease is scarce; it is commonly believed to be due to witches' attacks or bad fate. Four therapeutic paths are taken: self-medication, traditional therapy, the church and the health centre. However lay perception, recourse to traditional treatments and self-medication only partially explain the long delays in diagnosis (on average 6 months); the main problem lies with health providers, particularly the lack of proper diagnostic capability. CONCLUSIONS: Diagnostic capabilities at health centre level need to be strengthened through training and supervision. Engaging with the population and the traditional healers would render health promotion messages on BU more relevant and culturally acceptable. PMID- 19563477 TI - Molecular analysis of knock down resistance (kdr) mutation and distribution of kdr genotypes in a wild population of Culex quinquefasciatus from India. AB - OBJECTIVE: To investigate the presence of knock down resistance (kdr) mutation, its frequency distribution in the principal vector of bancroftian filariasis, Culex quinquefasciatus from northeastern India, and to relate kdr genotypes with susceptibility and/or resistance to DDT and deltamethrin in this vectors. METHODS: Adult female mosquitoes were collected by aspiration from human dwellings in two villages, Benganajuli and Rikamari, and two military establishments, Field Units I and II. Insecticide susceptibility tests were performed following WHO methods with 4% DDT and 0.05% deltamethrin. Molecular identification of kdr mutation and genotyping of kdr locus was performed by allele-specific PCR (AS-PCR) and direct sequencing in a subset of samples. RESULTS: Mosquitoes were resistant to DDT and showed 11.9-41.2% mortality, whereas the knock down bioassay for deltamethrin suggests complete susceptibility to this insecticide in all study sites except Benganajuli. The result of AS-PCR confirmed the presence of three genotypes: susceptible (SS), resistant (RR) and heterozygous (SR) in the population. Genotype frequencies at kdr locus for DDT resistant individuals conformed with the Hardy-Weinberg proportion, whereas DDT and deltamethrin susceptible individuals differed significantly (P < 0.05). The efficacy of AS-PCR in detecting the correct genotype was not encouraging. CONCLUSIONS: This is the first report from India on kdr genotyping in C. quinquefasciatus, and it confirms the occurrence of kdr allele in this vector in northeastern India. This finding has serious implications for the filariasis control programmes in India. PMID- 19563478 TI - Antimicrobial activity of Hymenaea martiana towards dermatophytes and Cryptococcus neoformans. AB - The biological activity of crude extract and fractions of Hymenaea martiana was evaluated against a panel of human pathogenic fungi. The crude extracts and hydroalcoholic fractions (E) showed a high activity against Cryptococcus neoformans species complex isolates with MICs between 2 and 64 MUg ml(-1). The methanolic (C) and butanolic (D) fractions were the most active against Trichopyton rubrum, Trichopyton mentagrophytes and Microsporum canis with MICs between 8 and 256 MUg ml(-1). None of the extracts was active against the yeast Malassezia furfur, Malassezia obtusa and Malassezia sympodialis. PMID- 19563479 TI - Setting priorities for health interventions in developing countries: a review of empirical studies. AB - OBJECTIVE: To assess and summarize empirical studies on priority-setting in developing countries. METHODS: Literature review of empirical studies on priority setting of health interventions in developing countries in Medline and EMBASE (Ovid) databases. RESULTS: Eighteen studies were identified and classified according to their characteristics and methodological approaches. All studies were published after 1999, mostly between 2006 and 2008. Study objectives and methodologies varied considerably. Most studies identified sets of relevant criteria for priority-setting (17/18) and involved different stakeholders as respondents (11/18). Studies used qualitative (8/15) or quantitative (3/15) techniques, or combinations of these (4/15) to elicit preferences from respondents. In a few studies, respondents deliberated on results (3/18). A minority of studies (7/18) resulted in a rank ordering of interventions. CONCLUSIONS: This review has revealed an increase in the number of empirical studies on priority-setting in developing countries in the past decade. Methods for explicit priority-setting are developing, being reported and are verifiable and replicable and can potentially lead to solutions for ad hoc policy-making in health care in many developing countries. PMID- 19563480 TI - Elimination capacity of a TSE-model agent in the manufacturing process of Alphanate/Fanhdi, a human factor VIII/VWF complex concentrate. AB - The variant Creutzfeldt-Jakob disease (vCJD) is a transmissible spongiform encephalopathy (TSE), mainly present in the UK and is associated with the ingestion of bovine products affected with bovine spongiform encephalopathy. Manufacturers of biological products must investigate the ability of their production processes to remove TSE agents. We studied the purification steps in the manufacturing process of two FVIII/VWF concentrates (Alphanate) and Fanhdi in their ability to eliminate an experimental TSE-model agent. Hamster scrapie strain 263K brain-derived materials were spiked into samples of the solutions taken before various stages during its production: 3.5% polyethylene glycol (PEG) precipitation, heparin affinity chromatography and saline precipitation/final filtrations. PEG precipitation and affinity chromatography were studied both as isolated and combined steps. TSE agent removal was determined using a laboratory scale model representative of the industrial manufacturing process. The prion protein (PrP(Sc)) was measured with Western blot and TSE infectivity was measured with bioassay. Western blot results were in agreement with those obtained by bioassay, showing a significant removal capacity in the production process: 3.21 3.43 log(10) for the PEG precipitation; about 3.45 log(10) for the affinity chromatography; and around 2.0 log(10) for the saline precipitation plus final filtrations. PEG precipitation and heparin affinity chromatography were demonstrated to be two complementary TSE-model agent removal mechanisms with total removal being the sum of the two. An overall reduction factor of around 8 log(10) can be deduced. The tests from the production process of FVIII/VWF complex concentrates have demonstrated their potential for eliminating TSE agents. PMID- 19563481 TI - Lymphoid hyperplasia with ulnar nerve compression in a severe haemophilia B patient--case report. PMID- 19563482 TI - The use of animal pharmacokinetic data to evaluate human dosing. PMID- 19563483 TI - Pancreas transplant and incidental Meckel's diverticulum: not always a straightforward decision. AB - INTRODUCTION: Unexpected intraoperative findings are not rare in surgical practice. Meckel's diverticulum with a mass is one such example. There are only two previously reported cases of Meckel's in transplantation, and neither involved pancreas transplant. RESULTS AND DISCUSSION: We present a case report of novel surgical technique using a Meckel's diverticulectomy site for the duodeno enterostomy to managing the exocrine secretions of the transplanted pancreas. We also discuss management of Meckel's diverticulum. The patient tolerated the procedure without complication, and continues to have normal renal and pancreatic function without any gastrointestinal (GI) complaints. The excised Meckel's diverticulum contained both gastric and pancreatic tissue. CONCLUSION: Although uncertainty about the best management practices exists in the general surgery patient population, given the potential complications that may arise from Meckel's diverticulum, in transplant patients the Meckel's should be removed when encountered. The point of excision can safely be incorporated into other intestinal anastomoses. PMID- 19563484 TI - Prevalence and staging of chronic kidney disease in renal transplant recipients. AB - INTRODUCTION: Diagnosis and staging of chronic kidney disease (CKD) is important for management and prevention of renal disease progression. It is unclear whether K/DOQI guidelines of the National Kidney Foundation are applicable to diagnosis of CKD in renal transplant recipients (RTRs) and which method is most appropriate for estimating glomerular filtration. OBJECTIVES: To determine the prevalence and staging of CKD in RTRs, according to K/DOQI guidelines, and the prevalence of complications of CKD. SUBJECTS AND METHODS: This cross-sectional study included RTRs at least six months post-transplantation followed at a single out-patient service. The glomerular filtration rate (GFR) was estimated with two different equations: the MDRD equation (Modification of Diet in Renal Disease) with four variables (age, creatinine level, gender, and race) and the Cockcroft-Gault (CG) formula. Patients with GFR more than 60 mL/min/1.73 m2 were diagnosed with CKD only in the presence of renal damage (hematuria, proteinuria, or evidence of injury in renal biopsy). CKD staging was compared to the two equations and the prevalence of complications was determined. RESULTS: The study evaluated 241 RTRs (average age: 40.6 +/- 12.5 yr, 62.2% male; 4.5% black, 50.6% from cadaveric donors). Average follow-up time was 6.8 +/- 6.1 yr and the average baseline creatinine level was 1.48 +/- 0.72 mg/dL. CKD was diagnosed in 70.5% of RTRs, of whom 52.9% (MDRD)/47.6% (CG) were classified as Stage III (GFR: 30-59 mL/min/1.73 m2). The agreement between the two methods was very close with regard to CKD diagnosis (kappa = 0.92) and close for stage-dependent prevalence (kappa = 0.68). The prevalence of anemia, hypocalcemia, hyperphosphatemia (HF), hyperuricemia (HU), and systemic arterial hypertension (SAH) was 10.6%, 7.6%, 10.3%, 54%, and 73.4% for patients with CKD. Significant differences were observed for HU, HF and SAH in patients without CKD. Anemia, HU and SAH were associated with CKD stage (p < 0.001). CONCLUSION: The prevalence of CKD in the study population was high (70.5%). The two equations tested correlated closely when used for GFR estimation. Routine CKD staging in RTRs would provide patients with safer and more appropriate management. PMID- 19563485 TI - Assessment of cardiovascular risk in waiting-listed renal transplant patients: a single center experience in 558 cases. AB - Cardiac screening is recommended to prevent cardiovascular death after renal transplantation. This retrospective observational study illustrates the results of application of a cardiac assessment algorithm in a series of 558 renal transplant candidates at a single center in Turin, Italy. A dipyridamole-stress sestamibi myocardial scintiscan (DMS) performed in 302/558 (54.1%) cases was positive in 52 (17.2%), negative in 200 (66.2%), borderline in 16 (5.3%), and with signs of previous necrosis in 34 (11.4%). Coronary lesions detected by angiography in 48.1% of the 52 positives were treated medically (13.5%) or by percutaneous/surgical procedure (34.6%). Coronary lesions were detected in 14.1% of asymptomatic population subgroup. The minor and major cardiovascular event rates and the cardiovascular death rate were 1.9%, 0%, and 0%, respectively, in positive DMS group (high-cardiological risk) vs. 10%, 4.5%, and 3.5% in the negatives (p > 0.5; n.s.). It is suggested that not increased cardiovascular event or deaths rates in the high-risk group reflect early coronary lesion detection and correction. Since 55.9% of cardiovascular events or deaths occurred in the negative group more than 24 months after the DMS, its mandatory repetition every two yr after a negative finding is recommended. PMID- 19563486 TI - Reduced post-operative neutrophil activation in liver transplant recipients suffering from post-hepatitic cirrhosis. AB - BACKGROUND: It has been supposed that liver transplant recipients with hepatitis C virus infection have a higher incidence of infectious complications after transplantation. This study was designed to investigate whether neutrophil function is immediately affected by liver transplantation. METHODS: Biochemical values, plasma levels of myeloperoxidase (MPO), hydrogen peroxide production of neutrophils and neutrophil-platelet complexes were analyzed in 32 patients who underwent liver transplantation and 20 healthy volunteers. RESULTS: MPO levels were significantly increased 24 h after reperfusion. In post-hepatitic patients levels were significantly lower three d up to one wk post-transplant than in patients due to other liver diseases. One wk post-operatively the respiratory burst activity following N-formyl-methionyl-leucylphenylalanine (fMLP) or (tumor necrosis factor-alpha) TNF-alpha/fMLP stimulation was depressed in post-hepatitic recipients. Respiratory burst stimulated with phorbol 12-myristate 13-acetate in these patients was increased one wk after transplantation. One d after transplantation the neutrophil-platelet complexes decreased significantly throughout the post-operative period. CONCLUSIONS: The results of this study suggest a reduced post-operative neutrophil activation in liver transplant recipients suffering from post-hepatitic cirrhosis compared to cirrhosis due to other causes. We hypothesized that neutrophil dysfunction in those patients depends on the underlying disease with an increased susceptibility to bacterial or fungal infections. PMID- 19563487 TI - Disparity in utilization of combined kidney-liver transplantation in the United States. AB - BACKGROUND: Orthotopic liver transplantation (OLT) is performed as a definitive treatment of acute and chronic liver failure. The prevalence of acute and chronic kidney diseases is substantially higher in this population secondary to diverse etiologies. Combined kidney-liver transplantation (CKLT) is widely performed in some centers, even though there are no definitive studies which support or contradict this practice. METHODS: We comprehensively reviewed OLT as well as CKLT data from US transplant centers provided by United Network of Organ Sharing (UNOS). RESULTS: The incidence of CKLT as a percentage of total OLTs performed has been increasing, especially in the post-MELD era (2002 and after). Moreover, there is a great disparity among centers in regard to percentage of CKLTs to total OLTs. CONCLUSION: We conclude that there is much difference of opinion among US transplant centers as to indications for CKLT. A more scientific approach to this problem including studies to assess the role of kidney biopsy in determining renal outcome after OLT is needed. PMID- 19563488 TI - Non-HLA T-cell reactivity during the first year after HLA-identical living related kidney transplantation. AB - BACKGROUND: It has been reported that donor-reactive T-cell responses may decrease during the first year after HLA-mismatched organ transplantation. We wondered whether donor-reactive T-cell responses directed to minor histocompatibility antigens (mHAgs) or other non-HLA antigens also decrease after HLA-identical living-related (LR) kidney transplantation. METHODS: We studied donor-reactive T-cell responses by IFN-gamma and granzyme B (GrB) Elispot assays in 15 HLA-identical LR kidney transplant recipients before, six months and one yr after transplantation. Third-party reactivity was used as control. Patient and donor peripheral blood mononuclear cells were typed for 11 known mHAgs. RESULTS: During the study period, 60% and 36% of the patients demonstrated donor-reactive IFN-gamma and GrB producing cells (pc), respectively. The number of donor reactive IFN-gamma and GrB pc was significantly lower than the number of third party reactive IFN-gamma and GrB pc. After transplantation, donor-reactivity and third-party reactivity were comparable to pre-transplant values. No relation was found in mHAg mismatches between donor and recipient and donor-reactive T-cell response. CONCLUSIONS: Donor-reactivity could be detected before and after HLA identical LR kidney transplantation, but was not related with the number of mHAg mismatches, and did not decrease after transplantation. PMID- 19563489 TI - Susceptibility profile of vaginal isolates of Candida albicans prior to and following fluconazole introduction - impact of two decades. AB - Current treatment options for vulvovaginal candidiasis due to Candida albicans include over-the-counter and prescription antifungal agents. Fluconazole has been used extensively with an unknown impact on susceptibility. To investigate antifungal susceptibility trends in clinical vaginal isolates of C. albicans from 1986 to 2008, microdilution susceptibility was performed on randomly selected single isolates. Minimum inhibitory concentrations (MICs) were determined for: fluconazole, clotrimazole, miconazole, ketoconazole, itraconazole, voriconazole, flucytosine and amphotericin B. The MIC(90) for each drug was then calculated for the time periods: 1986-1989, 1992-1996 and 2005-2007. A total of 250 C. albicans vaginal isolates were included. The MIC(90) (mcg ml(-1) ) for fluconazole was 0.25, 0.5 and 0.5 mcg ml(-1) for each grouping, respectively. The corresponding MIC(90) for flucytosine was 1, 2 and 8 mcg ml(-1) , respectively. The MIC(90) for the remaining agents remained unchanged across time periods mentioned. Of note, the percentage of isolates with MIC >=1 and >=2 mcg ml(-1) for fluconazole increased from 3% to 9% over the study period. Although the C. albicans MIC(90) to fluconazole in vaginal isolates has not shown a clinically significant increase since 1986, there is an increasing number of isolates with elevated MICs. The implications of this increase are unknown, but given the achievable vaginal concentrations of fluconazole, reduced susceptibility may have clinical relevance. PMID- 19563490 TI - A 20-year survey of tinea faciei. AB - Tinea faciei is an uncommon dermatophytosis affecting the glabrous skin of the face. Between 1988 and 2007 at the Dermatology Department of Cagliari University, 107 cases of tinea faciei have been diagnosed, involving 72 females and 35 males, aged 2-72 years. Incidence peaks were observed between 6 and 15 years (48.59%) and between 36 and 45 years (17.76%). Males below and females above 15 years of age were the most affected. In 61 patients (57.1%), typical forms of tinea faciei were observed, whereas in 46 (42.9%), atypical forms were observed, mainly mimicking discoid lupus erythematosus (nine cases), and polymorphous light eruption (eight cases). Typical cases were present in younger patients, aged between 2 and 15 years, while atypical forms were distributed in any of the decades, but mostly between 36 and 72 years. Of the 46 cases of atypical presentation, 33 were females. The isolated dermatophytes were Microsporum canis (63 cases), Trichophyton rubrum (24 cases) and T. mentagrophytes var. mentagrophytes (20 cases). Seven males and two females aged 4-10 years were also affected by tinea capitis and eight patients (three males and five females) of various ages by tinea corporis. Eleven patients (two males and nine females) aged >35 years were affected by onychomycosis. All patients recovered after local and/or systemic antifungal therapy, without relapse or side effects. PMID- 19563491 TI - Influence of different susceptibility testing methods and media on determination of the relevant fluconazole minimum inhibitory concentrations for heavy trailing Candida isolates with low-high phenotype. AB - We investigated the incidence of trailing growth with fluconazole in 101 clinical Candida isolates (49 C. albicans and 52 C. tropicalis) and tried to establish the convenient susceptibility testing method and medium for fluconazole minimum inhibitory concentration (MIC) determination. MICs were determined by CLSI M27-A2 broth microdilution (BMD) and Etest methods on RPMI-1640 agar supplemented with 2% glucose (RPG) and on Mueller-Hinton agar supplemented with 2% glucose and 0.5 MUg ml(-1) methylene blue (GMB). BMD and Etest MICs were read at 24 and 48 h, and susceptibility categories were compared. All isolates were determined as susceptible with BMD, Etest-RPG and Etest-GMB at 24 h. While all isolates were interpreted as susceptible at 48 h on Etest-RPG and Etest-GMB, one C. albicans isolate was interpreted as susceptible-dose dependent (S-DD) and two C. tropicalis isolates were interpreted as resistant with BMD. On Etest-RPG, trailing growth caused widespread microcolonies within the inhibition zone and resulted in confusion in MIC determination. On Etest-GMB, because of the nearly absence of microcolonies within the zone of inhibition, MICs were evaluated more easily. We conclude that, for the determination of fluconazole MICs of trailing Candida isolates, the Etest method has an advantage over BMD and can be used along with this reference method. Moreover, GMB appears more beneficial than RPG for the fluconazole Etest. PMID- 19563492 TI - Re: Kerns MJJ, Darst MA, Olsen TG, Fenster M, Hall P, Grevey S. Shrinkage of cutaneous specimens: formalin or other factors involved? PMID- 19563493 TI - Increased HIF-1 alpha immunostaining in psoriasis compared to psoriasiform dermatitides. AB - BACKGROUND: Expression of hypoxia inducible factor 1 (HIF-1) alpha can be linked to inflammation through reciprocal interactions with several cytokines. This finding is in accordance with the previously noted HIF-1 alpha overexpression in psoriatic lesional keratinocytes. METHODS: In this study we have employed a carefully selected panel of antibodies and quantitative morphometric analysis to compare HIF-1 alpha immunoreactivity in psoriatic biopsy samples vs. that in samples obtained from psoriasiform dermatitides. RESULTS: We found statistically significant HIF-1 alpha overexpression in psoriasis. Furthermore, we observed a previously unreported preferential localization of immunoreactive keratinocytes in the immediate vicinity of inflamed and elongated papillae. This pattern of immunoreactivity may partially account for the observed increase of HIF-1 alpha immunostaining in psoriasis. CONCLUSIONS: Overall, our findings imply that possible novel therapeutic intervention(s) on the HIF-1 alpha pathway may offer another approach in controlling the inflammatory activity of psoriatic lesions. PMID- 19563494 TI - Solitary nodule of the penis as unique manifestation of Rosai-Dorfman disease. AB - We report the case of a 53-year-old man with a solitary nodule located on the inner surface of the prepuce. The lesion was removed and the final diagnosis was Rosai-Dorfman disease (RDD/sinus histiocytosis with massive lymphadenopathy). This report, being the first case of RDD presenting as a single nodule of the penis, broadens the spectrum of the cutaneous expression of RDD. PMID- 19563495 TI - Plexiform pattern in cutaneous granular cell tumors. AB - Granular cell tumors, considered to be of peripheral nerve sheath origin, can involve various parts of the body with skin and tongue being the most common organs involved. In the skin, the granular cell tumors usually form an ill defined mass of round to oval cells with abundant eosinophilic granular cytoplasm and central or eccentric bland nuclei. Here we present nine cases of granular cell tumors, which display features different from the classical cutaneous granular cell tumor and have features similar to the earlier described plexiform granular cell tumors. Plexiform pattern in granular cell tumors may be more common than is reported. PMID- 19563496 TI - Smoking, sun exposure, number of nevi and previous neoplasias are risk factors for melanoma in older patients (60 years and over). AB - BACKGROUND: Malignant melanoma risk factors have been studied in different geographical area populations. However, no study has focused on risk factors which are more frequently associated to the over 60's age group. METHODS: A case control study was performed that included 160 patients age > or = 60 years diagnosed of cutaneous melanoma and 318 controls matched for age and sex. Both groups were assessed, by personal interview and physical examination, for different phenotype characteristics (hair and eye color, phototype), the presence of other cutaneous lesions (solar lentigines, actinic keratoses and nevi), degree and type of solar exposure and personal and family past history of cutaneous or non-cutaneous cancer. Differences were evaluated by contingency tables and univariate and multivariate logistic regression. RESULTS: Of 17 factors, those risk factors with a strong effect on the development of melanoma in the elderly were: fair eyes, severe sunburns, years of occupational sun exposure, smoking, > 50 melanocytic nevi and personal history of NMSC and other non-cutaneous neoplasias. CONCLUSIONS: Tobacco smoking is an independent risk factor for cutaneous melanoma in the elderly. Intense (both acute and chronic) sun exposure and constitutional features, such as tumor susceptibility (NMSC, non-cutaneous neoplasias, and multiple nevi) are also associated with melanoma risk. All these factors should help to better design educational campaigns in older people. PMID- 19563497 TI - Improvements in patient-reported outcomes in moderate-to-severe psoriasis patients receiving continuous or paused etanercept treatment over 54 weeks: the CRYSTEL study. AB - OBJECTIVE: To assess patient-reported outcomes (PRO) in patients with moderate-to severe plaque psoriasis receiving continuous or paused etanercept treatment. METHODS: In a multicentre European open-label study, one group (n = 352) received continuous therapy: 25 mg subcutaneously (SC) twice weekly (BIW) throughout 54 weeks. The other group (n = 359) received paused therapy: 50 mg SC BIW (9.6), with the higher bleeding tendency having a higher score. Whole blood collected by phlebotomy and contact pathway suppressed by 100 microg mL(-1) CTI was stimulated to react by the addition of 5 pM TF. Reactions were quenched at 20 min by inhibitors. Thrombin generation, determined by enzyme-linked immunosorbent assay for thrombin-antithrombin was evaluated in terms of clot time (CT), maximum level (MaxL) and maximum rate (MaxR) and compared to the bleeding score. Data are shown as the mean+/-SD. MaxL was significantly different (P < 0.001) between the groups: 504 +/- 114, 315 +/- 117 and 194 +/- 91 nM; with higher thrombin concentrations in the groups with lower bleeding scores. MaxR was higher in the groups with a lower bleeding score; 97 +/- 51, 86 +/- 60 and 39 +/- 16 nM min(-1) (P = 0.09). No significant difference was detected in CT among the groups, 5.6 +/ 1.3, 4.7 +/- 0.7 and 5.6 +/- 1.3 min. Our empirical study in CTI-inhibited whole blood shows that the MaxL of thrombin generation appears to correlate with the bleeding phenotype of haemophilia A. PMID- 19563499 TI - Risk factors for inhibitor formation in haemophilia: a prevalent case-control study. AB - Inhibitor formation is a major complication of haemophilia treatment. In a prevalent case-control study, we evaluated blood product exposure, genotype and HLA type on haemophilia A inhibitor formation. Product exposure was extracted from medical records. Genotype was determined on stored DNA samples by detection of virtually all mutations-SSCP (DOVAM-S) and subcycling PCR. HLA typing was performed by PCR amplification and exonuclease-released fluorescence. Cases experienced higher intensity factor, 455 vs. 200 U per exposure, P < 0.005, more frequent central nervous system (CNS) bleeding, seven of 20 (35.0%) vs. one of 57 (1.7%), P = 0.001 and more commonly from inhibitor families, seven of 20 (35.0%) vs. zero of 57 (0%), P < 0.001, and African-American, 12 of 63 (19.0%) vs. six of 117 (5.1%), P = 0.015. Among the latter, CNS bleeding was more commonly the initial bleed, 60% vs. 0%, P < 0.001, and survival was shorter, 14 vs. 38 yr, P = 0.025. Inhibitor formation was uncommon in those with missense mutations, two of 65 (3.1%) vs. 31 of 119 (26.0%), P = 0.008, and unrelated to factor VIII immunogenic epitope, P = 0.388, or HLA type, P > 0.100. Genotype was not associated with race. Time to immune tolerance was shorter for titres <120 vs. > or = 120 BU/mL, six vs. 16 months, P < 0.01, but unaffected by tolerizing dose regimen, P > 0.50. Inhibitor formation is associated with high intensity product exposure, CNS bleeding, African-American race and low frequency of missense mutations. The ideal time to initiate prophylaxis to reduce CNS bleeding and inhibitor formation will require prospective studies. PMID- 19563503 TI - Insulin resistance is a major determinant of liver stiffness in nondiabetic patients with HCV genotype 1 chronic hepatitis. AB - BACKGROUND: In patients with chronic hepatitis C (CHC), liver stiffness measurement (LSM) by transient elastography (TE), is closely related to the stage of fibrosis, but may be affected by necroinflammation. Other factors, such as insulin resistance (IR), might influence the performance of LSM. AIMS: To evaluate in a cohort of nondiabetic patients with genotype 1 CHC, whether IR and other anthropometric, biochemical, metabolic and histological factors contribute to LSM and to identify the best cut-off values of LSM for predicting different stages of fibrosis. METHODS: Nondiabetic patients with genotype 1 CHC (n = 156) were evaluated by liver biopsy (Metavir score), anthropometric, biochemical and metabolic features including IR. Furthermore, all subjects underwent LSM by TE. RESULTS: Severe fibrosis (F3-F4) was associated with LSM (OR 1.291; 95%CI 1.106 1.508). LSM was also independently correlated with low platelets (P = 0.03), high gammaGT (P < 0.001) and high HOMA (P = 0.004) levels. A stiffness value > or =8 KPa was identified as the best cut-off for predicting severe fibrosis (AUC 0.870); yet this cut-off still failed to rule out F3-F4 fibrosis in 22.7% of patients (false-negative rate) or rule in F3-F4 in 19.6% (false-positive rate). Platelets <200 x 10(3)/mmc and a HOMA of >2.7 were the major determinants of these diagnostic errors in predicting severe fibrosis. Conclusions In nondiabetic patients with genotype 1 CHC, insulin resistance, gammaGT and platelet levels contribute to LSM independently of liver fibrosis. The identification of these three factors contributes to a more correct interpretation of LSM. PMID- 19563502 TI - Health care seeking for abdominal bloating and visible distention. AB - BACKGROUND: While knowledge has accumulated regarding health care seeking in several functional gastrointestinal disorders (FGIDs), little is known about health care seeking in those with bloating and distention. AIM: To identify predictors of health care seeking for bloating and distention. METHODS: The validated Talley Bowel Disease Questionnaire was mailed to a cohort selected at random from the population of Olmsted County, Minnesota; 2259 subjects (53% females; mean age 62 years) answered questions about bloating and distention. The complete medical record of each respondent was reviewed. Logistic regression was used to compare consulting for bloating and distention with consulting for other GI symptoms, and nonconsulters. RESULTS: A total of 131 (6%) subjects in the community consulted a physician for bloating or distention. Older age [odds ratio (OR), 1.8; 95% confidence interval (CI): 1.5, 2.1], higher somatic symptom scores (OR, 2.0; CI: 1.4, 2.8), lower education level (OR, 2.7; CI: 1.2, 5.6), early satiety (OR, 2.0; CI: 1.1, 3.8) and abdominal pain (OR, 2.4; CI: 1.6, 3.7) were associated with people seeking health care for bloating or distention vs. non consulters. Similarly, older age (OR, 1.4; CI: 1.2, 1.7), chronic constipation (OR, 2.0; CI: 1.2, 3.2) and visible distention (OR, 3.0; CI: 1.8, 4.9) had greater odds of presenting for bloating or distention compared with presenting for other GI symptoms; somatic symptoms were not a predictor (OR, 1.1; CI: 0.8, 1.5). CONCLUSIONS: Factors that lead people to present for bloating and distention are similar to those for other GI symptoms visits; however, specific biological rather than somatic features may predict visits for bloating and distention. PMID- 19563504 TI - The prognostic significance of histological features in oral squamous cell carcinoma. AB - BACKGROUND: Different factors predict nodal metastasis, recurrence and survival in oral cancer. The aim was to assess the prognostic value of histological features related to the primary tumour. METHODS: A total of 144 patients surgically treated at Odense University Hospital for oral cancer between 1999 and 2004 were included in the study. Postoperative radiation therapy was given in case of close and involved margins or high TNM stages (UICC 1997). Median follow up time was 38 months. All surgical resections were reviewed and 22 histological characteristics were assessed. RESULTS: The predominant sites were floor of mouth (FOM, 39%) and lateral tongue (35%). Fifty-nine per cent had UICC97 stage I-II disease. Five-year cause-specific survival was observed in 65%. Nodal involvement at diagnosis was observed in 36% which was significantly related to grade, neural and vascular invasion; surgical margins and increasing tumour depth. A cut-off value of 2 mm (4 mm for FOM) separated patients without and with nodal metastasis at the time of diagnosis. However, on multivariate analysis, neck disease was only associated with tumour depth and grade. Cox analysis of local recurrence in the oral cavity over time showed that tumour diameter and surgical margins were significant predictors while cause-specific survival was related to diameter, depth of invasion, surgical margins and extracapsular spread (ECS). CONCLUSIONS: Tumour depth and grade were strong prognostic factors for nodal metastasis, independently of other histological features. Tumour diameter and margins independently predict local recurrences in the oral cavity as well as cause specific survival. Nodal involvement and ECS were associated with adverse prognosis. PMID- 19563505 TI - Angiogenic and lymphangiogenic microvessel density in recurrent pleomorphic adenoma. AB - BACKGROUND: Recurrent pleomorphic adenoma (RPA) is an uncommon and challenging disease. The aim of this study was to determine if there is a difference between RPA and the pleomorphic adenoma (PA) without recurrence related to tumor blood and lymphatic vascularization. Moreover, we compared the microvessel density (MVD) between cell-rich areas (predominance of epithelial cells) and cell-poor areas (predominance of myxoid and chondroid areas) of the stroma of PA and RPA. In addition, immunohistochemical staining for the Ki-67 antigen was conducted simultaneously to evaluate cell proliferation in PA and RPA. METHODS: A total of 19 cases of PA and 24 cases of RPA, blood, and lymphatic vessels were analyzed by immunohistochemical technique using the antibodies CD34, CD105, D2-40, and Ki-67. RESULTS: Comparing no recurrent with recurrent tumor, no significant difference was found in terms of lymphatic vessel density, MVD, and proliferation index. When MVD and proliferation index were compared with different areas in cellular composition (cell-rich and cell-poor areas), there was a significant difference in PA, as well as in RPA. CONCLUSION: This study shows that although RPA presents more aggressive clinical behavior than PA, there is no difference between tumor blood and lymphatic vascularization, suggesting that there is no correlation between vascularity and risk of recurrence. Furthermore, vascularized stroma in PA, as well as RPA, depends on the proportion of the cellular composition. PMID- 19563507 TI - Morel's laminar sclerosis showing apraxia of speech: distribution of cortical lesions in an autopsy case. AB - A 57-year old man with chronic alcoholism presented with apraxia of speech and disturbance of consciousness. He had a history of gastrectomy and had been drinking alcohol. The symptoms improved with administration of thiamine, but he later developed diarrhea and delirium, and died approximately 40 days after the onset. Autopsy findings were consistent with Wernicke's encephalopathy and pellagra encephalopathy. Furthermore, laminar cortical necrosis with vacuoles and astrocytosis was found in the second and third layers of the bilateral frontal cortices, suggesting Morel's laminar sclerosis. The lesions were mainly located in the bilateral primary motor cortices. Involvement of the lower part of the left primary motor cortex may be associated with apraxia of speech in our case. PMID- 19563506 TI - Embryonal tumor with abundant neuropil and true rosettes: morphological, immunohistochemical, ultrastructural and molecular study of a case showing features of medulloepithelioma and areas of mesenchymal and epithelial differentiation. AB - Embryonal tumors are a group of malignant neoplasms that most commonly affect the pediatric population. Embryonal tumor with abundant neuropil and true rosettes is a recently recognized rare tumor. It is composed of neurocytes and undifferentiated neuroepithelial cells arranged in clusters, cords and several types of rosettes in a prominent neuropil-rich background. We describe a new case of this tumor. The patient, a 24-month-old female infant, was referred to the Meyer Children's Hospital with a history of right brachio-crural deficit associated with occasional episodes of headache and vomiting. Computed tomography scan and MRI revealed a large bihemispheric mass. The patient underwent two consecutive surgeries. The resultant surgical resection of the tumor was macroscopically complete. The postoperative period was uneventful. On light microscopy the tumor showed a composite morphology: embryonal tumor with abundant neuropil and true rosettes (specimen from the first surgery); medulloepithelioma with mesenchymal and epithelial areas (specimen from the second surgery). The immunohistochemistry evidenced the heterogeneous (neuronal, mesenchymal and epithelial) immunoprofile of tumoral cells. By real-time polymerase chain reaction (RT-PCR), the PTEN gene expression in the tumor was lower than in the five non-neoplastic brain tissues used as control. Mutation analysis did not show any variation in INI-1 and PTEN sequence while P53 analysis showed the presence of homozygote P72R variation. Fluorescent in situ hybridization analysis showed polysomy of chromosome 2 while amplification of N-MYC was not detected. Owing to the rarity of embryonal tumor with abundant neuropil and true rosettes, each new case should be recorded to produce a better clinical, pathological and molecular characterization of this lesion. PMID- 19563508 TI - Usefulness of smears in intra-operative diagnosis of newly described entities of CNS. AB - The recent edition of World Health Organisation (WHO) Classification of Tumours of the Central Nervous System has incorporated a substantial number of important changes. It has recognised several new entities, many of which are rare. Intra operative diagnosis of these tumours can be difficult with the freezing artefact that often cripples brain frozen sections. In many instances intra-operative smears are extremely useful adjuvants in neuropathological diagnosis. In this article, we describe intra-operative smear findings of three of the newly described tumours. Their characteristic cytologic features are illustrated along with differentiating features from the common mimics, together with a general approach to brain smears. The entities we discuss here are papillary glioneuronal tumour, papillary tumour of the pineal region and angiocentric glioma. All three tumours share at least focal pseudo-papillary/vasculocentric architecture. Smears from papillary glioneuronal tumour demonstrated dual population of cells in a neuropil background, whereas papillary tumour of the pineal region and angiocentric glioma comprise a single population of cells. These two tumours can further be differentiated based on their cell morphology and background. PMID- 19563510 TI - Effects of a 6-month infliximab treatment on plasma levels of leptin and adiponectin in patients with rheumatoid arthritis. AB - Patients with rheumatoid arthritis (RA) appear to have increased plasma levels of leptin and adiponectin. These adipokines may be implicated in the pathophysiology of RA. Tumour necrosis factor alpha (TNF-alpha) is a potential modulator of adipokines. The effects of long-term anti-TNF treatment on plasma levels of leptin and adiponectin are not clear. The aim of this study was to assess the effects of 6-month anti-TNF treatment (infliximab) on leptin and adiponectin plasma levels in RA patients. Thirty women with RA were included in the study. Patients with diabetes mellitus, any endocrine disorder or receiving any hypolipidemic or antidiabetic medication were not included. Thirty healthy age- and body mass index-matched women served as controls. Plasma levels of leptin and adiponectin were measured with enzyme immunoassay methods prior to and after the 6-month treatment with infliximab. Mean age and disease duration of patients were 51.8 +/- 14.4 and 12.2 +/- 6.7 years, respectively. Body weight did not change significantly over the 6-month period. Plasma levels of leptin and adiponectin were higher in patients than controls and did not change significantly after 6 month treatment. Interestingly, in the tertile of patients with the highest baseline adiponectin concentrations, adiponectin levels were significantly reduced (P < 0.05). Infliximab treatment did not change plasma levels of leptin and adiponectin after 6-month treatment in the whole study population. However, a reduction of adiponectin levels was observed in patients with higher baseline adiponectin levels. PMID- 19563509 TI - Early metabolic reactivation versus antioxidant therapy after a traumatic spinal cord injury in adult rats. AB - Disability after traumatic spinal cord injury (TSCI) results from physical trauma and from "secondary mechanisms of injury" such as low metabolic energy levels, oxidative damage and lipid peroxidation. In order to prove if early metabolic reactivation is a better therapeutic option than antioxidant therapy in the acute phase of TSCI, spinal cord contusions were performed in adult rats using a well characterized weight drop technique at thoracic 9 level. After TSCI, pyrophosphate of thiamine or non-degradable cocarboxylase (NDC) enzyme was used to maintain energy levels, antioxidants such as superoxide dismutase and catalase (ANT) were used to decrease oxidative damage and methylprednisolone (MP), which has both therapeutic properties, was used as a control. Rats were divided into one sham group and six with TSCI; one of them received no treatment, and the rest were treated with NDC, MP, NDC + MP, NDC + ANT or ANT. The ANT group decreased lactate and creatine phosphokinase levels and increased the amount of preserved tissue (morphometric analysis) as well as functional recovery (Basso, Beattie and Bresnahan or BBB motor scale). In contrast, NDC treatment increased lipid peroxidation, measured through thiobarbituric acid reactive substances (TBARS) levels, as well as spinal cord tissue destruction and functional deficit. Early metabolic reactivation after a TSCI may be deleterious, while natural early metabolic inhibition may not be a "secondary mechanism of injury" but a "secondary neuroprotective response". While increased antioxidant defence after a TSCI may currently be an ideal therapeutic strategy, the usefulness of metabolic reactivation should be tested in the sub-acute or chronic phases of TSCI and new strategies must continue to be tested for the early ones. PMID- 19563512 TI - Doppler-defined pulmonary hypertension and the risk of death in children with sickle cell disease followed for a mean of three years. AB - Pulmonary hypertension (PH) is associated with increased mortality in adults with sickle cell disease (SCD), but its prognostic significance in children is unknown. Eighty-eight children with SCD were followed after echocardiographic screening for PH. After a mean follow-up of 3 years, all 18 subjects with PH were alive. In our children, as in adults with SCD, PH was associated with increased haemolysis. In contrast, our subjects with PH did not have overt systemic disease observed in adults. PH may be a manifestation of progressive organ damage from chronic haemolysis and systemic vasculopathy that ultimately leads to early death in adulthood. PMID- 19563514 TI - Adrenal Hodgkin lymphoma. PMID- 19563513 TI - Abnormal centrosome-centriole cycle in chronic myeloid leukaemia? AB - Abnormal numbers, structures and functions of centrosomes in chronic myeloid leukaemia (CML) may influence cell proliferation and genomic instability, which are features of the disease. Centrosomes are regulators of mitotic spindle orientation and can act as scaffolds for centrosome-associated regulators of the cell cycle. This study showed, for the first time, that p210(BCR-ABL1) and p145(ABL1) are both centrosome-associated proteins, as demonstrated by co immunoprecipitation with the pericentriolar protein, pericentrin. Furthermore, when CML cells were treated with imatinib there was a 55% and 20% reduction of p210(BCR-ABL1) and p145(ABL1) binding to pericentrin, respectively. Cell lines expressing p210(BCR-ABL1) and primary CD34(+) cells from CML patients exhibited more numerical and structural centrosomal abnormalities than p210(BCR-ABL1) negative cells. Primary cells from CML blast crisis (BC) patients exhibited a distinctive amorphous staining pattern of pericentrin compared to normal and CML chronic phase (CP) patients, suggesting a possible defect in pericentrin localisation at the centrosomes. Proteins, such as aurora kinases, pericentrin, survivin and separase, regulate centrosome structure and function, cell cycle and mitotic spindle formation. Levels of the protease, separase are abnormally high in CML CP and BC cells in comparison to normal CD34(+) cells. The data imply that expression of p210(BCR-ABL1) is associated with abnormalities in the centrosome centriole cycle and increased separase expression. PMID- 19563515 TI - DRD1 5'UTR variation, sex and early infant stress influence ethanol consumption in rhesus macaques. AB - The mesolimbic dopamine system plays an important role in mediating a variety of behaviors and is involved in mediating the reinforcing effects of ethanol. Genes encoding dopamine receptor subtypes are thus good candidate loci for understanding the genetic etiologies of susceptibility to alcohol dependence and its antecedent behavioral phenotypes. We tested whether variation in DRD1 influences alcohol consumption in rhesus macaques and whether its influence is mediated by sex and early rearing experience. We genotyped a single nucleotide polymorphism (-111 G/T) in the 5'UTR of DRD1 in 96 subjects raised with their mothers until 6 months of age (n = 43) or in peer-only groups (n = 53). As young adults they underwent a 7-week voluntary ethanol consumption experiment. anova revealed a significant main effect of sex (F(1,95) = 6.3, P = 0.014) and an interaction between genotype, sex and rearing on ethanol consumption (F(7,95) = 4.63, P = 0.0002). Maternally deprived males heterozygous for the T allele consumed significantly more ethanol (P > t 4-2[[6 amino-9-(N-ethyl-b-D-ribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzene propanoic acid hydrochloride>2-chloro-N(6)-cyclopentyladenosine>-2-chloro-6-[[(3 iodophenyl)methyl]amino]-9H-purin-9-yl]-1-deoxy-N-methyl-b-D-ribofuranuronamide = adenosine. 4-(-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5 ylamino]ethyl) phenol, an A(2A) antagonist, reduced the relaxations to EFS, adenosine and NECA. In urothelium-intact samples, relaxations to EFS and purines were smaller than in urothelium-denuded preparations. Neuronal voltage-gated Na(+) channels blockade failed to modify ATP relaxations. At basal tension, EFS- and ATP-induced contractions were resistant to desensitization or blockade of P2X(1) and P2X(3) receptors. CONCLUSIONS AND IMPLICATIONS: ATP is involved in the non-adrenergic, non-cholinergic, non-nitrergic inhibitory neurotransmission in the pig bladder neck, producing relaxation largely through muscle A(2A) receptors after breakdown to adenosine, and P2Y(1) receptors after breakdown to ADP. Antagonists of these receptors may be useful for urinary incontinence treatment produced by intrinsic sphincteric deficiency. PMID- 19563529 TI - The phospholipase A(2) inhibitor methyl indoxam suppresses diet-induced obesity and glucose intolerance in mice. AB - BACKGROUND AND PURPOSE: Previous results have shown that mice lacking in the group 1B phospholipase A(2) (Pla2g1b) are resistant to obesity and diabetes induced by feeding a diabetogenic high-fat/high-carbohydrate diet. This study examined the potential of using the Pla2g1b inhibitor methyl indoxam as therapy to suppress diet-induced obesity and diabetes. EXPERIMENTAL APPROACH: Male C57BL/6 mice were fed the diabetogenic diet with or without methyl indoxam supplementation. Body weight gain, fasting plasma glucose levels, glucose tolerance and postprandial lysophospholipid absorption were compared. KEY RESULTS: Wild-type C57BL/6 mice fed the diabetogenic diet without Pla2g1b inhibitor showed 31 and 69% body weight gain after 4 and 10 weeks respectively. These animals also showed elevated plasma glucose levels and were glucose intolerant. In contrast, C57BL/6 mice fed the diabetogenic diet with 90 mg.kg(-1) of methyl indoxam gained only 5% body weight after 10 weeks. These animals were also euglycaemic and displayed normal glucose excursion rates in glucose tolerance test. Methyl indoxam suppression of diet-induced body weight gain and glucose intolerance was correlated with the inhibition of Pla2g1b-mediated postprandial lysophospholipid absorption. CONCLUSIONS AND IMPLICATIONS: These results show that oral supplementation of a diabetogenic diet with the Pla2g1b inhibitor methyl indoxam effectively suppresses diet-induced obesity and diabetes in mice. This suggests that Pla2g1b inhibition may be a potentially effective oral therapeutic option for treatment of obesity and diabetes. PMID- 19563527 TI - Dinucleoside polyphosphates: strong endogenous agonists of the purinergic system. AB - The purinergic system is composed of mononucleosides, mononucleoside polyphosphates and dinucleoside polyphosphates as agonists, as well as the respective purinergic receptors. Interest in the role of the purinergic system in cardiovascular physiology and pathophysiology is on the rise. This review focuses on the overall impact of dinucleoside polyphosphates in the purinergic system. Platelets, adrenal glands, endothelial cells, cardiomyocytes and tubular cells release dinucleoside polyphosphates. Plasma concentrations of dinucleoside polyphosphates are sufficient to cause direct vasoregulatory effects and to induce proliferative effects on vascular smooth muscle cells and mesangial cells. In addition, increased plasma concentrations of a dinucleoside polyphosphate were recently demonstrated in juvenile hypertensive patients. In conclusion, the current literature accentuates the strong physiological and pathophysiological impact of dinucleoside polyphosphates on the cardiovascular system. PMID- 19563530 TI - Osteoblasts play key roles in the mechanisms of action of strontium ranelate. AB - BACKGROUND AND PURPOSE: Strontium ranelate reduces fracture risk in postmenopausal women with osteoporosis. Evidence from non-clinical studies and analyses of bone markers in phase III trials indicate that this is due to an increase in osteoblast formation and a decrease of osteoclastic resorption. The aim of this work was to investigate, in human cells, the mechanisms by which strontium ranelate is able to influence the activities of osteoblasts and osteoclasts. EXPERIMENTAL APPROACH: Human primary osteoblasts were used to examine effects of strontium ranelate on replication (thymidine incorporation), differentiation (Runx2 and alkaline phosphatase) and cell survival (cell counts and caspase activity). Osteoprotegerin (OPG) was measured by quantitative reverse transcription PCR (qRT-PCR) and elisa and receptor activator of NFkappaB ligand (RANKL) by qRT-PCR and Western blot. As strontium ranelate has been proposed as an agonist of the calcium-sensing receptor (CaSR), the involvement of CaSR in the effects of strontium ranelate on OPG and RANKL expression, and cell replication was examined using siRNA. KEY RESULTS: Strontium ranelate increased mRNA and protein levels of OPG and suppressed those of RANKL. Strontium ranelate also stimulated osteoblast replication and differentiation and increased cell survival under stress. Knocking down CaSR suppressed strontium ranelate-induced stimulation of OPG mRNA, reduction of RANKL mRNA, and increase in replication, indicating the involvement of CaSR in these responses. CONCLUSIONS AND IMPLICATIONS: Our results demonstrate that osteoblasts play a key role in the mechanism of action of the anti-fracture agent, strontium ranelate by mediating both its anabolic and anti-resorptive actions, at least in part, via activation of CaSR. PMID- 19563532 TI - Pals-associated tight junction protein functionally links dopamine and angiotensin II to the regulation of sodium transport in renal epithelial cells. AB - BACKGROUND AND PURPOSE: Dopamine inhibits renal cell Na(+),K(+)-ATPase activity and cell sodium transport by promoting the internalization of active molecules from the plasma membrane, whereas angiotensin II (ATII) stimulates its activity by recruiting new molecules to the plasma membrane. They achieve such effects by activating multiple and distinct signalling molecules in a hierarchical manner. The purpose of this study was to investigate whether dopamine and ATII utilize scaffold organizer proteins as components of their signalling networks, in order to avoid deleterious cross talk. EXPERIMENTAL APPROACH: Attention was focused on a multiple PDZ domain protein, Pals-associated tight junction protein (PATJ). Ectopic expression of PATJ in renal epithelial cells in culture was used to study its interaction with components of the dopamine signalling cascade. Similarly, expression of PATJ deletion mutants was employed to analyse its functional relevance during dopamine-, ATII- and insulin-dependent regulation of Na(+),K(+) ATPase. KEY RESULTS: Dopamine receptors and components of its signalling cascade mediating inhibition of Na(+),K(+)-ATPase interact with PATJ. Inhibition of Na(+),K(+)-ATPase by dopamine was prevented by expression of mutants of PATJ lacking PDZ domains 2, 4 or 5; whereas the stimulatory effect of ATII and insulin on Na(+),K(+)-ATPase was blocked by expression of PATJ lacking PDZ domains 1, 4 or 5. CONCLUSIONS AND IMPLICATIONS: A multiple PDZ domain protein may add functionality to G protein-coupled and tyrosine kinase receptors signalling during regulation of Na(+),K(+)-ATPase. Signalling molecules and effectors can be integrated into a functional network by the scaffold organizer protein PATJ via its multiple PDZ domains. PMID- 19563531 TI - A new class of organic nitrates: investigations on bioactivation, tolerance and cross-tolerance phenomena. AB - BACKGROUND AND PURPOSE: The chronic use of organic nitrates is limited by serious side effects including oxidative stress, nitrate tolerance and/or endothelial dysfunction. The side effects and potency of nitroglycerine depend on mitochondrial aldehyde dehydrogenase (ALDH-2). We sought to determine whether this concept can be extended to a new class of organic nitrates with amino moieties (aminoalkyl nitrates). EXPERIMENTAL APPROACH: Vasodilator potency of the organic nitrates, in vitro tolerance and in vivo tolerance (after continuous infusion for 3 days) were assessed in wild-type and ALDH-2 knockout mice by isometric tension studies. Mitochondrial oxidative stress was analysed by L-012 dependent chemiluminescence and protein tyrosine nitration. KEY RESULTS: Aminoethyl nitrate (AEN) showed an almost similar potency to glyceryl trinitrate (GTN), even though it is only a mononitrate. AEN-dependent vasodilatation was mediated by cGMP and nitric oxide. In contrast to triethanolamine trinitrate (TEAN) and GTN, AEN bioactivation did not depend on ALDH-2 and caused no in vitro tolerance. In vivo treatment with TEAN and GTN, but not with AEN, induced cross tolerance to acetylcholine (ACh)-dependent and GTN-dependent relaxation. Although all nitrates tested induced tolerance to themselves, only TEAN and GTN significantly increased mitochondrial oxidative stress in vitro and in vivo. CONCLUSIONS AND IMPLICATIONS: The present results demonstrate that not all high potency nitrates are bioactivated by ALDH-2 and that high potency of a given nitrate is not necessarily associated with induction of oxidative stress or nitrate tolerance. Obviously, there are distinct pathways for bioactivation of organic nitrates, which for AEN may involve xanthine oxidoreductase rather than P450 enzymes. PMID- 19563533 TI - Autocrine amplification loop in statin-induced apoptosis of human melanoma cells. AB - BACKGROUND AND PURPOSE: Beside their cholesterol lowering effect, statins exert pleiotropic effects, which include anti-inflammatory, immunosuppressive and anti proliferative actions. In higher concentrations, statins trigger apoptosis in primary cells and tumour cells. In particular, melanoma cells have been found to be susceptible to statin-induced apoptosis, although only after longer incubation times. The molecular mechanisms behind this delayed drug-induced apoptosis are still unclear. EXPERIMENTAL APPROACH: The human melanoma A375 and 518A2 cell lines were exposed to various statins in a time-dependent and dose-dependent manner, and indicators of apoptosis, caspase activity and individual apoptotic pathways were analysed for 3-hydroxy-3-methylglutaryl-coenzyme A reductase dependent and independent effects. KEY RESULTS: Kinetic analysis of statin induced apoptosis revealed an apoptotic burst for exposure times longer than 24 h. While the extrinsic pathway was not activated within 24 h, longer incubation times corroborated amplification of the mitochondrial pathway with significant activation of caspase 8. Continuous refreshing of the simvastatin-containing medium abrogated the mitochondrial amplification loop via caspase 8. Moreover, conditional medium, supplemented with mevalonic acid in order to nullify a possible contamination by statins, significantly triggered caspase 8 activity. Fas ligand was excluded as a possible candidate to account for the statin-induced autocrine amplification loop. CONCLUSIONS AND IMPLICATIONS: Simvastatin and atorvastatin are capable of triggering an 'autocrine' suicide factor, which amplifies apoptosis via the extrinsic pathway in human melanoma cells. This pro apoptotic stimulus implies possible therapeutic potential and may guide feasibility for more potent statins in anti-cancer strategies. PMID- 19563534 TI - Analysis of binding site for the novel small-molecule TLR4 signal transduction inhibitor TAK-242 and its therapeutic effect on mouse sepsis model. AB - BACKGROUND AND PURPOSE: TAK-242, a novel synthetic small-molecule, suppresses production of multiple cytokines by inhibiting Toll-like receptor (TLR) 4 signalling. In this study, we investigated the target molecule of TAK-242 and examined its therapeutic effect in a mouse sepsis model. EXPERIMENTAL APPROACH: Binding assay with [(3)H]-TAK-242 and nuclear factor-kappaB reporter assay were used to identify the target molecule and binding site of TAK-242. Bacillus calmette guerin (BCG)-primed mouse sepsis model using live Escherichia coli was used to estimate the efficacy of TAK-242 in sepsis. KEY RESULTS: TAK-242 strongly bound to TLR4, but binding to TLR2, 3, 5, 9, TLR-related adaptor molecules and MD 2 was either not observed or marginal. Mutational analysis using TLR4 mutants indicated that TAK-242 inhibits TLR4 signalling by binding to Cys747 in the intracellular domain of TLR4. TAK-242 inhibited MyD88-independent pathway as well as MyD88-dependent pathway and its inhibitory effect was largely unaffected by lipopolysaccharide (LPS) concentration and types of TLR4 ligands. TAK-242 had no effect on the LPS-induced conformational change of TLR4-MD-2 and TLR4 homodimerization. In mouse sepsis model, although TAK-242 alone did not affect bacterial counts in blood, if co-administered with ceftazidime it inhibited the increases in serum cytokine levels and improved survival of mice. CONCLUSIONS AND IMPLICATIONS: TAK-242 suppressed TLR4 signalling by binding directly to a specific amino acid Cys747 in the intracellular domain of TLR4. When co administered with antibiotics, TAK-242 showed potent therapeutic effects in an E. coli-induced sepsis model using BCG-primed mice. Thus, TAK-242 may be a promising therapeutic agent for sepsis. PMID- 19563536 TI - Induction of a reversible, non-cytotoxic S-phase delay by resveratrol: implications for a mechanism of lifespan prolongation and cancer protection. AB - BACKGROUND AND PURPOSE: Resveratrol (RES) has been shown to prolong lifespan and prevent cancer formation. At present, the precise cellular mechanisms of RES actions are still not clearly understood, and this is the focus of this study. EXPERIMENTAL APPROACH: Using human hepatocellular carcinoma-derived HepG2 cells as a model, we studied RES-induced changes in cell growth, cell cycle progression and apoptosis. KEY RESULTS: RES at lower concentrations induced a strong but reversible S-phase delay and mild DNA synthesis inhibition, yet without causing apoptotic or necrotic cell death. At high concentrations, RES induced apoptosis, which is mainly mediated by the mitochondrial pathway. Overall, RES was a relatively weak apoptotic agent. Mechanistically, MEK inhibition was identified as an important early signalling event for RES-induced apoptosis. In comparison, activation of CDK2 and checkpoint kinase 2, and inhibition of phosphatidylinositol 3'-kinase/Akt signalling pathway contributed to the induction by RES of a reversible, non-cytotoxic S-phase delay. CONCLUSION AND IMPLICATIONS: It is hypothesized that the induction of a non-cytotoxic S-phase delay may represent a useful mechanistic strategy for lifespan prolongation and cancer prevention. When cell cycles are selectively slowed down in the S phase, it would cumulatively increase the total lifespan of an organism if the total numbers of cell divisions of a given organism are assumed to remain basically constant. Likewise, when cells proceed through the cell cycles at a reduced pace during DNA replication, it may allow cells more time to repair the damaged DNA, and thereby reduce the chances for mutagenesis and tumour initiation. PMID- 19563535 TI - Sphingolipids in inflammation: pathological implications and potential therapeutic targets. AB - Sphingolipids are formed via the metabolism of sphingomyelin, a constituent of the plasma membrane, or by de novo synthesis. Enzymatic pathways result in the formation of several different lipid mediators, which are known to have important roles in many cellular processes, including proliferation, apoptosis and migration. Several studies now suggest that these sphingolipid mediators, including ceramide, ceramide 1-phosphate and sphingosine 1-phosphate (S1P), are likely to have an integral role in inflammation. This can involve, for example, activation of pro-inflammatory transcription factors in different cell types and induction of cyclooxygenase-2, leading to production of pro-inflammatory prostaglandins. The mode of action of each sphingolipid is different. Increased ceramide production leads to the formation of ceramide-rich areas of the membrane, which may assemble signalling complexes, whereas S1P acts via high affinity G-protein-coupled S1P receptors on the plasma membrane. Recent studies have demonstrated that in vitro effects of sphingolipids on inflammation can translate into in vivo models. This review will highlight the areas of research where sphingolipids are involved in inflammation and the mechanisms of action of each mediator. In addition, the therapeutic potential of drugs that alter sphingolipid actions will be examined with reference to disease states, such as asthma and inflammatory bowel disease, which involve important inflammatory components. A significant body of research now indicates that sphingolipids are intimately involved in the inflammatory process and recent studies have demonstrated that these lipids, together with associated enzymes and receptors, can provide effective drug targets for the treatment of pathological inflammation. PMID- 19563538 TI - Growing pains and the brain pathology stimulus package. PMID- 19563539 TI - Protein aggregate myopathies. Introduction. PMID- 19563540 TI - Myofibrillar myopathies: a clinical and myopathological guide. AB - Myofibrillar myopathies (MFMs) are histopathologically characterized by desmin positive protein aggregates and myofibrillar degeneration. Because of the marked phenotypic and pathomorphological variability, establishing the diagnosis of MFM can be a challenging task. While MFMs are partly caused by mutations in genes encoding for extramyofibrillar proteins (desmin, alphaB-crystallin, plectin) or myofibrillar proteins (myotilin, Z-band alternatively spliced PDZ-containing protein, filamin C, Bcl-2-associated athanogene-3, four-and-a-half LIM domain 1), a large number of these diseases are caused by still unresolved gene defects. Although recent years have brought new insight into the pathogenesis of MFMs, the precise molecular pathways and sequential steps that lead from an individual gene defect to progressive muscle damage are still unclear. This review focuses on the clinical and myopathological aspects of genetically defined MFMs, and shall provide a diagnostic guide for this numerically significant group of protein aggregate myopathies. PMID- 19563537 TI - Drug-induced QT interval shortening: potential harbinger of proarrhythmia and regulatory perspectives. AB - ATP-dependent potassium channel openers such as pinacidil and levcromakalim have long been known to shorten action potential duration and to be profibrillatory in non-clinical models, raising concerns on the clinical safety of drugs that shorten QT interval. Routine non-clinical evaluation of new drugs for their potential to affect cardiac repolarization has revealed that drugs may also shorten QT interval. The description of congenital short QT syndrome in 2000, together with the associated arrhythmias, suggests that drug-induced short QT interval may be proarrhythmic, and an uncanny parallel is evolving between our appreciation of the short and the long QT intervals. Epidemiological studies report an over-representation of short QT interval values in patients with idiopathic ventricular fibrillation. Therefore, as new compounds that shorten QT interval are progressed further into clinical development, questions will inevitably arise on their safety. Arising from the current risk-averse clinical and regulatory environment and concerns on proarrhythmic safety of drugs, together with our lack of a better understanding of the clinical significance of short QT interval, new drugs that substantially shorten QT interval will likely receive an unfavourable regulatory review unless these drugs fulfil an unmet clinical need. This review provides estimates of parameters of QT shortening that may be of potential clinical significance. Rufinamide, a recently approved anticonvulsant, illustrates the current regulatory approach to drugs that shorten QT interval. However, to further substantiate or confirm the safety of these drugs, their approval may well be conditional upon large-scale post-marketing studies with a focus on cardiac safety. PMID- 19563541 TI - Inclusion body myositis: a degenerative muscle disease associated with intra muscle fiber multi-protein aggregates, proteasome inhibition, endoplasmic reticulum stress and decreased lysosomal degradation. AB - Sporadic inclusion body myositis (s-IBM), the most common muscle disease of older persons, is of unknown cause, and there is no enduring treatment. Abnormal accumulation of intracellular multi-protein inclusions is a characteristic feature of the s-IBM phenotype, and as such s-IBM can be considered a "conformational disorder," caused by protein unfolding/misfolding combined with the formation of inclusion bodies. Abnormal intracellular accumulation of unfolded proteins may lead to their aggregation and inclusion body formation. The present article is focusing on the multiple proteins that are accumulated in the form of aggregates within s-IBM muscle fibers, and it explores the most recent research advances directed toward a better understanding of mechanisms causing their impaired degradation and abnormal aggregation. We illustrate that, among other factors, abnormal misfolding, accumulation and aggregation of proteins are associated with their inadequate disposal-and these factors are combined with, and perhaps provoked by, an aging intracellular milieu. Other concurrent and possibly provocative phenomena known within s-IBM muscle fibers are: endoplasmic reticulum stress and unfolded protein response, mitochondrial abnormalities, proteasome inhibition, lysosome abnormality and endodissolution. Together, these appear to lead to the s-IBM-specific vacuolar degeneration, and muscle fiber atrophy, concluding with muscle fiber death. PMID- 19563542 TI - Extralysosomal protein degradation in myofibrillar myopathies. AB - Myofibrillar myopathies (MFMs) are a group of heterogeneous muscle disorders morphologically defined by the presence of foci of dissolution of the myofibrils, accumulation of the products of myofibrillar degradation and ectopic expression of multiple proteins. MFMs represent the paradigm of conformational protein diseases of skeletal and cardiac muscles. Protein aggregation in MFMs is now considered to be the result of a failure of the extralysosomal proteolytic degradation system. Several factors including mutant proteins, aggresome formation and oxidative stress may compromise the ubiquitin-proteasome system, promoting the accumulation of potentially toxic protein aggregates within muscle cells. PMID- 19563543 TI - Actinopathies and myosinopathies. AB - The currently recognized two forms of "anabolic" protein aggregate myopathies, that is, defects in development, maturation and final formation of respective actin and myosin filaments encompass actinopathies and myosinopathies. The former are marked by mutations in the ACTA1 gene, largely of the de novo type. Aggregates of actin filaments are deposited within muscle fibers. Early clinical onset is often congenital; most patients run a rapidly progressive course and die during their first 2 years of life. Myosinopathies or myosin storage myopathies also commence in childhood, but show a much more protracted course owing to mutations in the myosin heavy chain gene MYH7. Protein aggregation consists of granular material in muscle fibers and few, if any, filaments. PMID- 19563544 TI - 5-year-old boy with a clival mass. PMID- 19563545 TI - 31-year-old man with balint's syndrome and visual problems. PMID- 19563546 TI - 5-year old male with an interhemispheric frontal mass. PMID- 19563547 TI - 64-year-old man with multiple cerebral lesions. PMID- 19563548 TI - A female patient with a right leg paresis. PMID- 19563549 TI - A 69-year-old man with left hemispheric necrotizing mass lesion. PMID- 19563552 TI - The intra-uterine environment and placentation. PMID- 19563554 TI - Substance use and violent death: a case for the 'too hard' basket? PMID- 19563555 TI - First steps first. PMID- 19563556 TI - Controlling gambling: a population-based perspective to measurement and monitoring as resource for effective interventions. PMID- 19563553 TI - Vascular dysfunction in the diabetic placenta: causes and consequences. AB - The development and functioning of the human fetoplacental vascular system are vulnerable to the maternal diabetic milieu. These vessels are in direct continuum with the fetal vascular system and are therefore also vulnerable to fetal endocrine derangements. Increased angiogenesis, altered junctional maturity and molecular occupancy, together with increased leakiness, constitute a well described phenotype of vessels in the Type 1 diabetic human placenta and can be related to increased levels of placental vascular endothelial growth factor. The causes of these observed changes, whether maternal hyperglycaemia or fetal hyperinsulinaemia, still remain to be shown in the human placenta. Mechanistic studies using different vascular systems have shown high glucose and insulin to have profound vascular effects, with elevations in vascular endothelial growth factor, nitric oxide and protein kinase C being behind alterations in junctional adhesion molecules such as occludin and vascular endothelial-cadherin and vascular leakage of albumin. The role of advanced glycation products and oxidative stress in this vascular pathology is also discussed. The altered molecular mechanisms underlying the vascular changes in the diabetic human placenta may reflect similar consequences of high glucose and hyperinsulinaemia. PMID- 19563557 TI - Problem gambling: we should measure harm rather than 'cases'. PMID- 19563558 TI - Temperance, alcohol, and the American evangelical: a reassessment. AB - Abstinence from alcohol is a way of life for many American evangelicals, with rates of abstention running at over 70% among some Pentecostal denominations. This paper examines the religious beliefs that, historically, have supported teetotalism. The most notable of these is Christian perfection, a doctrine that originated in 18th-century England, that was then radicalized in America in the early 19th century. Abstinence from alcohol is highest among denominations that make Christian perfection the cornerstone of their teachings, and lowest among those that discount human agency. The paper also argues that 19th-century American evangelicals were by no means committed uniformly to temperance as a way of life, and that this was especially true of the various Methodist churches. PMID- 19563559 TI - Feasibility and validity of low-budget telephonic follow-up interviews in routine outcome monitoring of substance abuse treatment. AB - AIMS: Routine outcome monitoring (ROM) is receiving growing attention. However, follow-up interviews are time-consuming and costly. This study examines the feasibility and validity of low-budget telephonic follow-up interviews for ROM in a substance abuse treatment centre (SATC). DESIGN: Observational study using data collected for routine outcome monitoring. SETTING: The study was performed in a SATC in an urban area in the Netherlands. Participants Feasibility and validity were assessed on data of 2325 patients. MEASUREMENTS: Data on pre-treatment socio demographic and clinical characteristics were collected using electronic patient records (EPRs) and the European version of the Addiction Severity Index (EuropASI). Data on intensity of treatment were also collected through the EPRs. Telephonic follow-up interviews were conducted between 9 and 10 months after intake. FINDINGS: A 53% follow-up rate was achieved; 35% of the patients could not be contacted, 3% explicitly refused and in 8% other reasons accounted for non participation. About 50% of the interviews took place in the intended time-frame. Costs were Euro 40 (57 US dollars) per completed interview. There were indications of selection bias, because patients with cocaine as their primary problem and patients with polysubstance abuse were under-represented in the follow-up sample; the presence of these disorders is associated with negative treatment outcome. CONCLUSIONS: Implementing telephonic low-budget follow-up interviews for ROM is feasible, but selection bias threatens internal validity of data, limiting generalization to the total treatment population. Increased efforts to track patients for follow-up may improve generalization. PMID- 19563560 TI - Comments on surveying alcohol in Africa. PMID- 19563561 TI - Alcohol advertising regulation: where to from here? PMID- 19563562 TI - Comparing retention in treatment and mortality in people after initial entry to methadone and buprenorphine treatment. AB - AIM To compare retention in treatment and mortality among people entering methadone and buprenorphine treatment for opioid dependence. DATA SOURCES: The Pharmaceutical Drugs of Abuse System (PHDAS) database records start- and end dates of all episodes of methadone and buprenorphine treatment in New South Wales, and the National Death Index (NDI) records all reported deaths. METHODS: Data linkage study. First entrants to treatment between June 2002 and June 2006 were identified from the PHDAS database. Retention in treatment was compared between methadone and buprenorphine. Names were linked to the NDI database, and 'good matches' were identified. Deaths were classified as occurring during induction, maintenance and either post-methadone or post-buprenorphine, depending on the latest episode of treatment prior to death. The numbers of inductions into treatment, of total person-years spent in each treatment, and person-years post methadone or buprenorphine, were calculated. Risk of death in different periods, and different treatments, was analysed using Poisson regression. RESULTS: A total of 5992 people entered their first episode of treatment-3349 (56%) on buprenorphine, 2643 on methadone. Median retention was significantly longer in methadone (271 days) than buprenorphine (40 days). During induction, the risk of death was lower for buprenorphine (relative risk = 0.114, 95% confidence interval = 0.002-0.938, P = 0.02, Fisher's exact test). Risk of death was lowest during treatment, significantly higher in the first 12 months after leaving both methadone and buprenorphine. Beyond 12 months after leaving treatment, risk of death was non-significantly higher than during treatment. CONCLUSIONS: Buprenorphine was safer during induction. Despite shorter retention in treatment, buprenorphine maintenance was not associated with higher risk of death. PMID- 19563563 TI - The longitudinal association between multiple substance use discrepancies and marital satisfaction. AB - AIMS: The objective of this work was to examine the relation between patterns of substance use among newly married couples and marital satisfaction over time. In particular, this work examined if differences between husbands' and wives' heavy alcohol use and cigarette smoking, rather than simply use per se, predicted decreases in marital satisfaction over the first 7 years of marriage. METHODS: Married couples (n = 634 couples) were assessed on a variety of substance use and relationship variables at the time of marriage and again at the first, second, fourth and seventh years of marriage. RESULTS: After controlling for key socio demographic variables, discrepancies in husband and wife cigarette smoking and heavy alcohol use were related to significant reductions in marital satisfaction. Importantly, couples who were discrepant on both substances experienced the greatest declines in marital satisfaction over time. CONCLUSIONS: Patterns of substance use among newly married couples are important predictors of changes in marital functioning over time. It was not simply the heavy alcohol use or cigarette smoking that predicted dissatisfaction but, rather, differences between husbands' and wives' substance use that impacted the relationship. PMID- 19563565 TI - Database linkage: outside reflections on health care inside prisons. PMID- 19563564 TI - A social network perspective on heroin and cocaine use among adults: evidence of bidirectional influences. AB - AIMS: While several studies have documented a relationship between initiation of drug use and social network drug use in youth, the direction of this association is not well understood, particularly among adults or for stages of drug involvement beyond initiation. The present study sought to examine two competing theories (social selection and social influence) in the longitudinal relationship between drug use (heroin and/or cocaine) and social network drug use among drug experienced adults. DESIGN: Three waves of data came from a cohort of 1108 adults reporting a life-time history of heroin and/or cocaine use. SETTING: Low-income neighborhoods with high rates of drug use in Baltimore, Maryland. PARTICIPANTS: Participants had weekly contact with drug users and were 18 years of age or older. MEASUREMENTS: Drug use data were self-report. Network drug use was assessed through a social network inventory. Close friends were individuals whom the participant reported seeing daily or rated as having the highest level of trust. Findings Structural equation modeling indicated significant bidirectional influences. The majority of change in network drug use over time was due to change in the composition of the network rather than change in friends' behavior. Drug use by close peers did not influence participant drug use beyond the total network. CONCLUSIONS: There is evidence of both social selection and social influence processes in the association between drug use and network drug use among drug-experienced adults. PMID- 19563566 TI - A survey of tobacco dependence treatment guidelines in 31 countries. AB - AIMS: The Framework Convention on Tobacco Control (FCTC) asks countries to develop and disseminate comprehensive evidence-based guidelines and promote adequate treatment for tobacco dependence, yet to date no summary of the content of existing guidelines exists. This paper describes the national tobacco dependence treatment guidelines of 31 countries. DESIGN, SETTING, PARTICIPANTS: A questionnaire on tobacco dependence treatment guidelines was sent by e-mail to a convenience sample of contacts working in tobacco control in 31 countries in 2007. Completed questionnaires were received from respondents in all 31 countries. During the course of these enquiries we also made contact with people in 14 countries that did not have treatment guidelines and sent them a short questionnaire asking about their plans to produce guidelines. MEASUREMENTS: The survey instrument was a 17-item questionnaire asking the following key questions: do the guidelines recommend brief interventions, intensive behavioural support, medications; which medications; do the guidelines apply to the whole health-care system and all professionals; do they refer explicitly to the Cochrane database; are they based on another country's guidelines; are they national or more local; are they endorsed formally by government; did they undergo peer review; who funded them; where were they published; do they include evidence on cost effectiveness of treatment? FINDINGS: According to respondents, all their countries' guidelines recommended brief advice, intensive behavioural support and nicotine replacement therapy (NRT); 84% recommended bupropion; 19% recommended varenicline; and 35% recommended telephone quitlines. Nearly half (48%) included cost-effectiveness evidence. Seventy-one per cent were supported formally by their government and 65% were supported financially by the government. Most (84%) used the Cochrane reviews as a source of evidence, 84% underwent a peer review process and 55% were based on the guidelines of other countries, most often the United States and England. CONCLUSION: Overall, the guidelines reviewed followed the evidence base closely, recommending brief interventions, intensive behavioural support and NRT, and most recommended bupropion. Varenicline was not on the market in most of the countries in this survey when their guidelines were written, illustrating the need for guidelines to be updated periodically. None recommended interventions not proven to be effective, and some recommended explicitly against specific interventions (for lack of evidence). Most were peer reviewed, many through lengthy and rigorous procedures, and most were endorsed or supported formally by their governments. Some countries that did not have guidelines expressed a need for technical support, emphasizing the need for countries to share experience, something the FCTC process is well placed to support. PMID- 19563567 TI - Effect of isometric exercise and body scanning on cigarette cravings and withdrawal symptoms. AB - AIMS: To examine the acute effects of a guided relaxation routine (body scan) and isometric exercise on desire to smoke and tobacco withdrawal symptoms. DESIGN: Experimental comparison of three conditions. PARTICIPANTS: Forty-eight individuals reporting smoking > or =10 cigarettes daily. INTERVENTION: Random assignment to one of three interventions delivered via a 10-minute audio: isometric exercise (IE, n = 14), body scanning (BS, n = 18) or a reading about natural history (control group, n = 16). Interventions were delivered twice on the same day: in the laboratory, then in their 'normal' environment. MEASUREMENTS: Desire to smoke (primary outcome) and withdrawal symptoms were rated at pre-intervention and up to 30 minutes post-intervention. FINDINGS: Controlling for baseline scores, post-intervention desire to smoke and withdrawal symptoms were significantly lower for IE and BS groups, compared with the controls, in both environments. There were no significant differences for IE versus BS. For desire to smoke, controlling for baseline values, ratings in the laboratory were significantly lower for IE and BS versus the control up to 30 minutes post-intervention. In the normal environment, these ratings were significantly lower only up to 5 minutes post-intervention. CONCLUSIONS: Brief IE and BS interventions are effective for reducing desire to smoke and withdrawal symptoms in temporarily abstaining smokers. These interventions were found to be more effective in the laboratory than in the smoker's normal environment, but this may be an artefact of there not being a sufficient 'wash-out' period between interventions. These techniques may be beneficial for managing desire to smoke and tobacco withdrawal. PMID- 19563568 TI - Addiction medicine is an attractive field for young physicians-a call for a European initiative for the training in addiction medicine. PMID- 19563569 TI - 2008-Lithuania's year of sobriety: alcohol control becomes a priority of health policy. PMID- 19563572 TI - Vascular catheterization is difficult in infants with Down syndrome. PMID- 19563573 TI - Sinus arrest on the electrocardiograph: pseudo or real? PMID- 19563571 TI - Inhale, suction and close the lung: a common clinical practice in Scandinavian intensive care units? PMID- 19563575 TI - Failure to inflate Ambu AuraFlex single-use LMA after insertion. PMID- 19563576 TI - An innovative functional equivalent of the jet stylet. PMID- 19563577 TI - Fluid therapy monitors in acute infarction. PMID- 19563579 TI - Objective monitoring of neuromuscular block should become the standard of care. PMID- 19563580 TI - Incidence of severe hypoglycemia with tight glycemic control: the higher the better? PMID- 19563582 TI - The metabolism and pharmacokinetics of isotretinoin in patients with acne and rosacea are not influenced by ethanol. AB - BACKGROUND: Isotretinoin is effective in the treatment of severe acne and rosacea. Both parent drug and its main metabolite 4-oxo-isotretinoin are potentially teratogenic compounds and contain a carboxylic acid moiety. In the presence of ethanol, naturally occurring as well as synthetic retinoids also containing a carboxylic acid moiety are capable of undergoing an ethyl esterification with the metabolic formation of more lipophilic compounds with a much longer terminal half-life. OBJECTIVES: To determine if isotretinoin (13-cis RA), its main metabolite 4-oxo-isotretinoin (4-oxo-13-cis-RA), and other possible metabolites in the presence or absence of ethanol are converted to their corresponding ethyl derivatives in patients with severe acne or rosacea after multiple isotretinoin dosing. In addition, pharmacokinetic parameters of the parent drug and its 4-oxo metabolite were determined. PATIENTS/METHODS: Eleven patients with severe acne or rosacea were treated with isotretinoin daily for 3 months and investigated pharmacokinetically during 24 h after 1 month of treatment and for up to 28 days after discontinuation of therapy. A possible influence of ethanol was evaluated using a simple self-administered questionnaire and by measuring serum ethanol levels during treatment. The concentrations of isotretinoin, 4-oxo-isotretinoin and possible ethylated and nonethylated metabolites were measured by reverse-phase high-performance liquid chromatography. RESULTS: Although seven of 11 patients had a considerable weekly alcohol intake, no endogenous synthesis of ethyl derivatives of isotretinoin, the main 4-oxo metabolite or the all-trans compounds was chromatographically detectable in any of the patients' plasma samples during the treatment period. Multiple dose pharmacokinetic data for the parent drug and its main metabolite were comparable to previous studies. CONCLUSIONS: The metabolism and pharmacokinetics of isotretinoin and its main metabolites are not influenced by ethanol during long-term isotretinoin treatment. After ceasing long-term isotretinoin therapy the recommended period of 1 month for using anticonceptive measures in fertile women seems adequate. PMID- 19563583 TI - Developmental disabilities: challenges for research practices and policies in the 21st century. PMID- 19563584 TI - Is levetiracetam a treatment option for dyskinetic cerebral palsy? PMID- 19563585 TI - Neurological manifestations of the Mendelian-inherited autoinflammatory syndromes. AB - Autoinflammatory syndromes include an expanding list of conditions characterized by unprovoked recurrent attacks of systemic inflammation with lack of auto antibodies or autoreactive T cells. Many of these syndromes are genetic diseases with a Mendelian inheritance. Neurological manifestations may be one of the major clinical features and, in some cases, the presenting symptom of these syndromes. The purpose of this review is to increase the recognition among neurologists of the Mendelian-inherited autoinflammatory syndromes by highlighting the neurological manifestations in the context of other symptoms that should lead physicians to suspect these syndromes. Most important for neurologists are the cryopyrin-associated periodic syndromes that include familial cold autoinflammatory syndrome, Muckle-Wells syndrome and neonatal-onset multisystem inflammatory disease (called chronic infantile neurological cutaneous and articular syndrome in Europe). We also review other syndromes with less common neurological involvement, including familial Mediterranean fever, tumor necrosis factor receptor-associated periodic syndrome, and hyperimmunoglobulinemia D syndrome. Because these syndromes are often treatable and irreversible damage is prevented if they are treated early, it is important to recognize the features that may result in these syndromes presenting to a neurologist, especially in early childhood. PMID- 19563586 TI - Long-term efficacy and tolerability of 4-monthly versus yearly botulinum toxin type A treatment for lower-limb spasticity in children with cerebral palsy. AB - In this study, we compared the long-term efficacy and tolerability of two dosage regimens of the potent botulinum toxin type A (BoNT-A; Dysport; Ipsen Ltd, Slough, UK) in children with cerebral palsy (CP) and lower-limb spasticity. Children aged 1 to 8 years with diplegic CP who were able to walk (aided or unaided) were randomized (1:1) to 30 LD(50) units/kg total body weight of BoNT-A (injected into gastrocnemius muscles) every 4 months or once yearly for 2 years in this multicentre, assessor-blinded, parallel-group study. In the 4-monthly group (n=110, 39 males, 71 females), mean age was 3 years 8 months (SD 1 y 6 mo, range 1-8 y). In the yearly group (n=104, 47 males, 57 females), mean age was 4 years 4 months (SD 1 y 6 mo, range 2-8 y). Both treatment groups had similar baseline Gross Motor Function Measure scores. At month 28 (primary endpoint; intention-to-treat group), median maximum passive ankle dorsiflexion was 12.00 degrees in the 4-monthly and 11.00 degrees in the yearly group. Between-group difference of 1.67 degrees was not statistically significant (p=0.055). Other efficacy endpoints showed no significant difference between the regimens. The results of the study do not allow a clear conclusion of the preferred injection regimen. PMID- 19563587 TI - Design and application of a screening and training protocol for odour testers in the field of personal care products. AB - The assessment of odours and in particular of human axillary odour is an integral part of the research and development of deodorant and anti-perspirant products. One method to perform odour assessment is the odour evaluation that is carried out by experts, designated as odour testers or sniffers. Product development decisions are therefore based on human assessment. As for every scientific measurement, the influencing factors need to be standardized or regularly calibrated as effectively as possible for reasons of quality assurance. We therefore developed a screening and training concept aiming to examine the general suitability of odour testers by determining the individual odour sensitivity for relevant odours. This newly developed method is based on the national and international standards and guidelines EN 13725:2003, VDI 3882 sheet 1 and ASTM-1207. Suitable odour testers are subsequently trained to correlate their individual odour intensity perception with an intensity calibration scale in order to achieve reproducible results. Training sessions held on a regular basis help to achieve a greater homology in the response of an existing panel. Our established screening and training protocol has already been successfully put into practice and is also subject to permanent improvement with regard to practical requirements. PMID- 19563588 TI - Antioxidant capacity of 3D human skin EpiDerm model: effects of skin moisturizers. AB - The objective of this study was to determine the effects of skin moisturizers on total antioxidant capacity (TAC) of human skin using EpiDerm model. Three different skin moisturizers containing antioxidant ingredients (samples 1-3) or aloe vera extract were topically applied to EpiDerm units and incubated for 2 and 24 h to determine acute and longer-term effects of applied samples on TAC and glutathione peroxidase activity in medium and/or homogenized skin tissues. Total antioxidant capacity in medium and skin homogenates was enhanced (P < 0.0001) by gel containing antioxidant ingredients (sample 2) after 2 and 24 h of incubation. Total antioxidant capacity in medium was also enhanced (P < 0.001) by cream containing antioxidant ingredients (sample 3) after 24 h of incubation. Overall, TAC in medium was greater (P < 0.02) after 24 h than 2 h of incubation. Skin moisturizer cream with high antioxidant levels determined by using oxygen radical absorbance capacity testing (sample 1) and aloe vera extract did not affect TAC. Glutathione peroxidase activity was enhanced (P < 0.0001) in medium and skin homogenates by sample 2 but not by any other sample. These data demonstrate high potential of gel and cream (samples 2 and 3) containing antioxidant ingredients in enhancing antioxidant capacity of EpiDerm which will likely contribute to overall skin health. Results of this experiment will help to better understand mechanisms of effects of skin moisturizers containing antioxidant ingredients on skin function at the tissue level and to establish effective strategies for skin protection and clinical treatments of skin disorders and possibly healing wounds. PMID- 19563589 TI - Combined structural and biological activities for new polyunsaturated fatty derivatives obtained by biotechnological process. AB - The objective of this study is to demonstrate the use of new polyunsaturated fatty derivatives for cutaneous applications. These new compounds present an analogue structure of cutaneous lipid, stabilize the polyunsaturated fatty acids (face to oxidation) and demonstrate specific biological activities. Three molecules described are Omega 6 fatty acid stabilized compound (O6FASC), the O3FASC and the O9FASC. The derivatives are synthesized via the same biotechnological process. This work describes the choice of final structure, the design of the biotechnological process and the free solvent enzymatic synthesis used for the synthesis of these three cutaneous lipid analogues. The restructuring effect of such analogues has been demonstrated with an in vivo study on volunteers. The stabilization of the O3FASC and O6FASC, and the biological activities of these three compounds are presented. The O6FASC shows very good results in anti-inflammatory effects; the O3FASC has anti-stress activities, whereas the O9FASC presents interesting results in improving elasticity and firmness. All these activity tests are presented in this work. PMID- 19563607 TI - Neutrophils: key mediators of tumour angiogenesis. AB - It is now well known that most malignant tumours contain a significant amount of leucocytic infiltrates the presence of which has, on many occasions, been linked to poor patient prognosis. These leucocyte populations are recruited to tumours by chemotactic factors released by either viable or necrotic tumour cells, or by cells within the tumour stroma. In recent times, most studies have analysed the role that tumour-associated macrophages (TAM) have on tumour progression. However, there is now increasing evidence to show that neutrophils also actively participate in this process. Whilst there are some data to suggest that neutrophil-derived factors can promote genetic mutations leading to tumourigenesis, or secrete factors that promote tumour cell proliferation; there is now substantial evidence to show that neutrophils, like TAM, significantly affect tumour angiogenesis. In this review, we discuss the likely mechanisms by which neutrophils are recruited into the tumour and then elaborate on how these cells may induce tumour vascularization by the secretion of powerful pro angiogenic factors. We also discuss possible future chemotherapeutic strategies that are aimed at limiting tumour angiogenesis by inhibiting neutrophil recruitment. PMID- 19563606 TI - A critical analysis of current in vitro and in vivo angiogenesis assays. AB - The study of angiogenesis has grown exponentially over the past 40 years with the recognition that angiogenesis is essential for numerous pathologies and, more recently, with the advent of successful drugs to inhibit angiogenesis in tumours. The main problem with angiogenesis research remains the choice of appropriate assays to evaluate the efficacy of potential new drugs and to identify potential targets within the angiogenic process. This selection is made more complex by the recognition that heterogeneity occurs, not only within the endothelial cells themselves, but also within the specific microenvironment to be studied. Thus, it is essential to choose the assay conditions and cell types that most closely resemble the angiogenic disease being studied. This is especially important when aiming to translate data from in vitro to in vivo and from preclinical to the clinic. Here we critically review and highlight recent advances in the principle assays in common use including those for endothelial cell proliferation, migration, differentiation and co-culture with fibroblasts and mural cells in vitro, vessel outgrowth from organ cultures and in vivo assays such as chick chorioallantoic membrane (CAM), zebrafish, sponge implantation, corneal, dorsal air sac, chamber and tumour angiogenesis models. Finally, we briefly discuss the direction likely to be taken in future studies, which include the use of increasingly sophisticated imaging analysis systems for data acquisition. PMID- 19563609 TI - The vasculature in rheumatoid arthritis: cause or consequence? AB - The expansion of the synovial lining of joints in rheumatoid arthritis (RA) necessitates an increase in the vascular supply to the synovium, to cope with the increased requirement for oxygen and nutrients. New blood vessel formation 'angiogenesis'- is recognized as a key event in the formation and maintenance of the pannus in RA, suggesting that targeting blood vessels in RA may be an effective future therapeutic strategy. Although many pro-angiogenic factors have been demonstrated to be expressed in RA synovium, vascular endothelial growth factor (VEGF) has been demonstrated to a have a central involvement in the angiogenic process in RA. Nevertheless, it is unclear whether angiogenesis - whether driven by VEGF and/or other factors - should be considered as a 'cause' or 'consequence' of disease. This ongoing 'chicken vs. egg' debate is difficult, as even the success of angiogenesis inhibition in models of RA does not provide a direct answer to the question. This review will focus on the role of the vasculature in RA, and the contribution of different angiogenic factors in promoting disease. Although no data regarding the effectiveness of anti angiogenic therapy in RA have been reported to date, the blockade of angiogenesis nevertheless looks to be a promising therapeutic avenue. PMID- 19563608 TI - Angiogenesis drives psoriasis pathogenesis. AB - Psoriasis pathogenesis is closely associated with disease-inducing Th1 and Th17 cells. Yet, several studies suggest that aberrant keratinocyte or endothelial cell signalling significantly contributes to disease manifestation. Histological hallmarks of psoriatic skin include the infiltration of multiple immune cells, keratinocyte proliferation and increased dermal vascularity. Formation of new blood vessels starts with early psoriatic changes and disappears with disease clearance. Several angiogenic mediators like vascular endothelial growth factor, hypoxia-inducible factors, angiopoietins and pro-angiogenic cytokines, such as tumour necrosis factor (TNF), interleukin (IL)-8 and IL-17, are up-regulated in psoriasis development. Contact- and mediator-dependent factors derived from keratinocytes, mast cells and immune cells may contribute to the strong blood vessel formation of psoriasis. New technologies and experimental models provide new insights into the role of angiogenesis in psoriasis pathogenesis. Interestingly, many therapies target not only immune cells, but also protein structures of endothelial cells. Here we summarize the role of pro-angiogenic factors in psoriasis development and discuss angiogenesis as a potential target of novel therapies. PMID- 19563611 TI - Microtubule depolymerizing vascular disrupting agents: novel therapeutic agents for oncology and other pathologies. AB - Vascular disrupting agents (VDAs) are a relatively new group of 'vascular targeting' agents that exhibit selective activity against established tumour vascular networks, causing severe interruption of tumour blood flow and necrosis to the tumour mass. Microtubule depolymerizing agents form by far the largest group of small molecular weight VDAs many of which, including lead compound disodium combretastatin A-4 3-O-phosphate (CA-4-P), are under clinical development for cancer. Although distinct from the angiogenesis inhibitors, VDAs can also interfere with angiogenesis and therefore constitute a potential group of novel drugs for the treatment of pathological conditions characterized by excessive angiogenesis, in addition to cancer. The endothelial cytoskeleton is the primary cellular target of this family of drugs, and some progress in understanding the molecular and signalling mechanisms associated with their endothelial disrupting activity has been made in the last few years. Susceptibility of tumour vessels to VDA damage is ascribed to their immature pericyte-defective nature, although the exact molecular mechanisms involved have not been clearly defined. Despite causing profound damage to tumours, VDAs fail to halt tumour growth unless used together with conventional treatments. This failure is attributed to resistance mechanisms, primarily associated with cells that remain viable within the tumour rim, and enhanced angiogenesis. The focus is now to understand mechanisms of susceptibility and resistance to identify novel molecular targets and develop strategies that are more effective. PMID- 19563610 TI - Coronary vessel development and insight towards neovascular therapy. AB - Formation of the coronary arteries consists of a precisely orchestrated series of morphogenetic and molecular events which can be divided into three distinct processes: vasculogenesis, angiogenesis and arteriogenesis (Risau 1997; Carmeliet 2000). Even subtle perturbations in this process may lead to congenital coronary artery anomalies, as occur in 0.2-1.2% of the general population (von Kodolitsch et al. 2004). Contrary to the previously held dogma, the process of vasculogenesis is not limited to prenatal development. Both vasculogenesis and angiogenesis are now known to actively occur within the adult heart. When the need for regeneration arises, for example in the setting of coronary artery disease, a reactivation of embryonic processes ensues, redeploying many of the same molecular regulators. Thus, an understanding of the mechanisms of embryonic coronary vasculogenesis and angiogenesis may prove invaluable in developing novel strategies for cardiovascular regeneration and therapeutic coronary angiogenesis. PMID- 19563612 TI - Can eccentric arterial plaques alone cause flow stagnation points and favour thrombus incorporation? AB - We have used an experimental model of aorta stenosis, with a Plexiglas plug, simulating a stable atheromatous plaque that promotes local turbulence and thrombosis. With animal survival of more than 24 h, we followed the partial fibrinolysis of the thrombus as well as its posterior organization and incorporation to the arterial wall as a neointima for up to 30 days. The mushroom plug form permitted the development of recirculation and stasis areas around it, favouring this evolution. Despite noted limitations, this study demonstrates that thrombus incorporation can contribute to plaque extension, as it can promote recirculation and stasis areas. PMID- 19563613 TI - High dietary methionine plus cholesterol stimulates early atherosclerosis and late fibrous cap development which is associated with a decrease in GRP78 positive plaque cells. AB - The role of homocysteine, or its precursor methionine, in the formation of fibrous caps and its association with endoplasmic reticulum (ER) stress is unclear. Homocysteine can stimulate collagen accumulation and upregulate the ER stress chaperone glucose regulated protein 78 (GRP78). The aim of this study was to determine if high dietary methionine would increase fibrous caps, and that removal of an atherogenic diet would decrease the amount of ER stressed cells. New Zealand white rabbits were fed for 2, 4, or 12 weeks an atherogenic diet [1% methionine + 0.5% cholesterol (2MC, 4MC or 12MC)]; for 4 or 12 weeks a 0.5% cholesterol diet (4Ch, 12Ch); and to study plaque regression, an MC diet for 2 or 4 weeks accompanied by 10 weeks of a normal diet (2MCr, 4MCr). Endothelial function, atherosclerosis and GRP78 positive cells were studied. Endothelial function was abolished in 4MC and atherosclerosis increased 17-fold (P < 0.05) compared with 4Ch. Fibrous caps composed 48% of total plaque area in 12MC vs. 10% in 12Ch (P < 0.01), and 12MC expressed less GRP78 plaque cells vs. 12Ch (P < 0.01). Four MCr had less plaque GRP78 cells than 12MC (P < 0.05) and less endothelial GRP78 cells (P < 0.01). In addition, GRP78 positive cells were the highest in 4MC, but decreased in all other groups (P < 0.01). GRP78 positive cells within the fibrous cap inversely correlated with cap size (r(2) = 0.9). These studies suggest that high dietary methionine could be beneficial for plaque stabilisation, and a normal diet also stabilises plaque and decreases the number of stressed plaque cells. PMID- 19563614 TI - A detailed microscopic study of the changes in the aorta of experimental model of postmenopausal rats fed with repeatedly heated palm oil. AB - Hypercholesterolaemia, increase in lipid peroxidation and hyperhomocysteinaemia may contribute to the pathogenesis of atherosclerosis. This study was performed to examine the effects of repeatedly heated palm oil mixed with 2% cholesterol diet on atherosclerosis in oestrogen-deficient postmenopausal rats. Ovariectomy causes disruption of tunica intima layer of the rat aorta simulating a postmenopausal condition in females. Twenty-four ovariectomized female Sprague Dawley rats were divided into four groups. The control group received 2% cholesterol diet without palm oil. A diet with 2% cholesterol content fortified with fresh, once-heated and five-times-heated palm oil was given to the other treatment groups. The rats were sacrificed at the end of 4 months of study and the aortic arch tissue was processed for histomorphometry and electron microscopy. On observation, there was disruption of the intimal layer of the ovariectomized rat aorta. There was no obvious ultrastructural change in the aorta of the rats fed with fresh palm oil. The ultrastructural changes were minimal with once-heated palm oil, in which there was a focal disruption of the endothelial layer. The focal disruption was more pronounced with five-times heated palm oil. The results of this study show that the ingestion of fresh palm oil may have a protective effect on the aorta but such a protective action may be lost when the palm oil is repeatedly heated. The study may be clinically important for all postmenopausal women who are susceptible to atherosclerosis. PMID- 19563615 TI - Apoptosis, cell proliferation and modulation of cyclin-dependent kinase inhibitor p21(cip1) in vascular remodelling during vein arterialization in the rat. AB - Neo-intima development and atherosclerosis limit long-term vein graft use for revascularization of ischaemic tissues. Using a rat model, which is technically less challenging than smaller rodents, we provide evidence that the temporal morphological, cellular, and key molecular events during vein arterialization resemble the human vein graft adaptation. Right jugular vein was surgically connected to carotid artery and observed up to 90 days. Morphometry demonstrated gradual thickening of the medial layer and important formation of neo-intima with deposition of smooth muscle cells (SMC) in the subendothelial layer from day 7 onwards. Transmission electron microscopy showed that SMCs switch from the contractile to synthetic phenotype on day 3 and new elastic lamellae formation occurs from day 7 onwards. Apoptosis markedly increased on day 1, while alpha actin immunostaining for SMC almost disappeared by day 3. On day 7, cell proliferation reached the highest level and cellular density gradually increased until day 90. The relative magnitude of cellular changes was higher in the intima vs. the media layer (100 vs. 2 times respectively). Cyclin-dependent kinase inhibitors (CDKIs) p27(Kip1) and p16(INKA) remained unchanged, whereas p21(Cip1) was gradually downregulated, reaching the lowest levels by day 7 until day 90. Taken together, these data indicate for the first time that p21(Cip1) is the main CDKI protein modulated during the arterialization process the rat model of vein arterialization that may be useful to identify and validate new targets and interventions to improve the long-term patency of vein grafts. PMID- 19563616 TI - Apoptosis and fibrosis are early features of heart failure in an animal model of metabolic cardiomyopathy. AB - In previous experiments, we observed signs of cardiac failure in mice overexpressing lipoprotein lipase (LPL) under the control of a muscle specific promotor and in peroxisome proliferators activated receptor alpha (PPARalpha) knockout mice overexpressing LPL under the control of the same promotor. In our current investigations, we focussed on morphological consequences and changes in mRNA and protein expression in hearts from these animals. mRNA expression was analysed by differential display analysis and Northern blot as well as by cDNA microarray analysis followed by pathway analysis. Protein expression was examined using immunoblot and immunohistochemistry. Fibrosis was determined by chromotrope aniline blue staining for collagen. A distinct increase in the expression of alpha-tubulin mRNA was noted in hearts of all mutant mouse strains compared with the control. This result was paralleled by increased alpha-tubulin protein expression. Using cDNA microarray analysis, we detected an activation of apoptosis, in particular an increase of caspase-3 expression in hearts of mice overexpressing LPL but not in PPARalpha knockout mice overexpressing LPL. This finding was confirmed immunohistochemically. In addition, we identified a distinct interstitial increase in collagen and an increase around blood vessels. In our mouse model, we detect mRNA and protein changes typical for cardiomyopathy even before overt clinical signs of heart failure. In addition, a small but distinct increase in the rate of apoptosis of cardiomyocytes and fibrotic changes contributes to cardiac failure in mice overexpressing LPL, whereas additional deficiency in PPARalpha seems to protect hearts from these effects. PMID- 19563617 TI - Expression of ADAM-15 in rat myocardial infarction. AB - A disintegrin and metalloprotease-15 (ADAM-15) is a potential novel regulator of inflammatory response and tissue remodelling, which is thought to have the ability to attenuate the cardiac function resulting from myocardial infarction (MI). Therefore, the aim of our study was to investigate the expression of ADAM 15 in rat MI. Wistar rats were subjected to MI by ligation of the left anterior descending coronary artery. Euthanasia was performed at 1, 3, 7 and 14 days following MI. The mRNA and protein expression levels of ADAM-15 were detected respectively by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The localization of ADAM-15 protein was observed by immunohistochemistry. Compared with sham-MI, the expression of ADAM-15 in MI increased at day 1, reached to maximum at day 3, decreased at day 7 and day 14 gradually. In addition, we also found that the localization of ADAM-15 was mainly at cardiac myocytes in the border area of MI and some macrophages in the border and infarcted areas. This study revealed a significant difference of ADAM-15 expression in rat MI and indicated that ADAM-15 maybe one of the important factors involved in inflammatory response and cardiac remodelling of rat MI. PMID- 19563620 TI - Acute heat stress brings down milk secretion in dairy cows by up-regulating the activity of the milk-borne negative feedback regulatory system. AB - BACKGROUND: The objective of this study was to determine if acute heat stress (HS) decreases milk secretion by activating the milk-borne negative feedback system, as an emergency physiological response to prevent a life-threatening situation. To induce HS, summer acclimatized dairy cows were exposed to full sun under mid-summer Mediterranean conditions, with and without conventional cooling procedures. RESULTS: Exposure to HS induced a rapid and acute (within 24 h) reduction in milk yield in proportion to the heat load. This decrease was moderated by cooler night-time ambient temperature. The reduction in milk yield was associated with corresponding responses in plasminogen activator/plasminogen plasmin activities, and with increased activity (concentration) of the (1-28) N terminal fragment peptide that is released by plasmin from beta-casein (beta-CN (1-28)). These metabolites constitute the regulatory negative feedback system. Previously, it has been shown that beta-CN (1-28) down-regulated milk secretion by blocking potassium channels on the apical aspects of the mammary epithelial cells. CONCLUSION: Here we demonstrate that the potassium channels in mammary tissue became more susceptible to beta-CN (1-28) activity under HS. Thus, the present study highlighted two previously unreported features of this regulatory system: (i) that it modulates rapidly in response to stressor impact variations; and (ii) that the regulations of the mammary epithelial potassium channel sensitivity to the inhibitory effect of beta-CN (1-28) is part of the regulatory system. PMID- 19563618 TI - Ischaemia/reperfusion induced cardiac stem cell homing to the injured myocardium by stimulating stem cell factor expression via NF-kappaB pathway. AB - Ischaemia/reperfusion (I/R) is a major cause of heart failure. Recently cardiac stem cells (CSCs) were proposed as the most appropriate cell type for heart disease therapy. However, it is still unclear whether I/R can stimulate the CSCs homing to the injured myocardium. Male Sprague-Dawley rats were subjected to a 30 min ischaemia followed by reperfusion of different intervals. RT-PCR, western blotting and immunohistochemistry were performed to detect stem cell factor (SCF) expression at mRNA and protein levels respectively. Activation of nuclear factor kappaB (NF-kappaB) was determined by electrophoretic mobility shift assay. To assess the homing of CSCs in vivo, BrdU-labelled CSCs were injected into AV groove before induction of ischaemia and examined by immunofluorescent staining in the injured myocardium after I/R. From day 3 to day 6 after reperfusion, the accumulation of CSCs was significantly elevated in the injured area, which was matched with the increased SCF expression during I/R. Pretreatment of rats with NF-kappaB inhibitor, N-acetyl-L-cysteine (NAC) not only suppressed NF-kappaB activation induced by I/R but also attenuated SCF expression. Further analysis revealed that I/R induced phosphorylation of IkappaBalpha after 15 min of reperfusion, and the raised phosphor-IkappaBalpha returned to the basal level at 2 h of reperfusion. In simulated I/R(SI/R) in vitro, it enhanced NF-kappaB activation and SCF expression in cultured neonatal rat cardiomyocytes, which was markedly inhibited by NF-kappaB decoy oligodeoxynucleotide or NAC. Taken together, our results demonstrated that I/R induced CSCs homing to the injured myocardium by stimulating myocardial SCF expression via activation of NF-kappaB. PMID- 19563621 TI - Training simulated patients: evaluation of a training approach using self assessment and peer/tutor feedback to improve performance. AB - BACKGROUND: Most medical schools use simulated patients (SPs) for teaching. In this context the authenticity of role play and quality of feedback provided by SPs is of paramount importance. The available literature on SP training mostly addresses instructor led training where the SPs are given direction on their roles. This study focuses on the use of peer and self evaluation as a tool to train SPs. METHODS: SPs at the medical school participated in a staff development and training programme which included a) self-assessment of their performance while observing video-tapes of their role play using a structured guide and b) peer group assessment of their performance under tutor guidance. The pre and post training performance in relation to authenticity of role play and quality of feedback was blindly assessed by students and tutors using a validated instrument and the scores were compared. A focus group discussion and a questionnaire assessed acceptability of the training programme by the SPs. RESULTS: The post training performance assessment scores were significantly higher (p < 0.05) than the pre-training scores. The degree of improvement in the quality of feedback provided to students was more when compared to the improvement of role play. The acceptability of the training by the SPs was very satisfactory scoring an average of 7.6 out of 10. The majority of the SPs requested the new method of training to be included in their current training programme as a regular feature. CONCLUSION: Use of structured self-reflective and peer-interactive, practice based methods of SP training is recommended to improve SP performance. More studies on these methods of training may further refine SP training and lead to improvement of SP performance which in turn may positively impact medical education. PMID- 19563623 TI - The epidemiology and survival of extrapulmonary small cell carcinoma in South East England, 1970-2004. AB - BACKGROUND: Extrapulmonary small cell carcinoma (EPSCC) is a rare cancer and few studies describe its epidemiology. Our objectives were to compare the incidence and survival of EPSCC in South East England with small cell carcinoma of the lung (SCLC), to determine the most common anatomical presenting sites for EPSCC and to compare survival in EPSCC by disease stage and site of diagnosis. METHODS: We used data from the Thames Cancer Registry database for South East England between 1970 and 2004 to determine the incidence, most common anatomical sites, and survival by site, and stage of EPSCC. 1618 patients registered with EPSCC were identified. We calculated the age-standardised incidence rate for EPSCC using the European standard population and compared this to that for SCLC. We calculated survival using the Kaplan-Meier method for EPSCC and SCLC, and reported 3-year survival for different EPSCC anatomical sites and disease stages. RESULTS: The incidence of EPSCC was much lower than for SCLC, similar in males and females, and stable throughout the study period, with incidence rates of 0.45 per 100,000 in males and 0.37 in females during 2000-2004. In general, patients with EPSCC had a better 3-year survival (19%) than SCLC (5%). The most common anatomical sites for EPSCC were oesophagus (18%), other gastrointestinal (15%), genitourinary (20%), head and neck (11%), and breast (10%). Breast EPSCC had the best 3-year survival (60%) and gastrointestinal EPSCC the worst (7%). CONCLUSION: This study suggests that EPSCC has a stable incidence and confirms that it presents widely, but most commonly in the oesophagus and breast. Site and extent of disease influence survival, with breast EPSCC having the best prognosis. Further studies using standardised diagnosis, prospective case registers for uncommon diseases and European cancer registries are needed to understand this disease. PMID- 19563622 TI - Seeking unique and common biological themes in multiple gene lists or datasets: pathway pattern extraction pipeline for pathway-level comparative analysis. AB - BACKGROUND: One of the challenges in the analysis of microarray data is to integrate and compare the selected (e.g., differential) gene lists from multiple experiments for common or unique underlying biological themes. A common way to approach this problem is to extract common genes from these gene lists and then subject these genes to enrichment analysis to reveal the underlying biology. However, the capacity of this approach is largely restricted by the limited number of common genes shared by datasets from multiple experiments, which could be caused by the complexity of the biological system itself. RESULTS: We now introduce a new Pathway Pattern Extraction Pipeline (PPEP), which extends the existing WPS application by providing a new pathway-level comparative analysis scheme. To facilitate comparing and correlating results from different studies and sources, PPEP contains new interfaces that allow evaluation of the pathway level enrichment patterns across multiple gene lists. As an exploratory tool, this analysis pipeline may help reveal the underlying biological themes at both the pathway and gene levels. The analysis scheme provided by PPEP begins with multiple gene lists, which may be derived from different studies in terms of the biological contexts, applied technologies, or methodologies. These lists are then subjected to pathway-level comparative analysis for extraction of pathway-level patterns. This analysis pipeline helps to explore the commonality or uniqueness of these lists at the level of pathways or biological processes from different but relevant biological systems using a combination of statistical enrichment measurements, pathway-level pattern extraction, and graphical display of the relationships of genes and their associated pathways as Gene-Term Association Networks (GTANs) within the WPS platform. As a proof of concept, we have used the new method to analyze many datasets from our collaborators as well as some public microarray datasets. CONCLUSION: This tool provides a new pathway-level analysis scheme for integrative and comparative analysis of data derived from different but relevant systems. The tool is freely available as a Pathway Pattern Extraction Pipeline implemented in our existing software package WPS, which can be obtained at http://www.abcc.ncifcrf.gov/wps/wps_index.php. PMID- 19563624 TI - Models of epidemics: when contact repetition and clustering should be included. AB - BACKGROUND: The spread of infectious disease is determined by biological factors, e.g. the duration of the infectious period, and social factors, e.g. the arrangement of potentially contagious contacts. Repetitiveness and clustering of contacts are known to be relevant factors influencing the transmission of droplet or contact transmitted diseases. However, we do not yet completely know under what conditions repetitiveness and clustering should be included for realistically modelling disease spread. METHODS: We compare two different types of individual-based models: One assumes random mixing without repetition of contacts, whereas the other assumes that the same contacts repeat day-by-day. The latter exists in two variants, with and without clustering. We systematically test and compare how the total size of an outbreak differs between these model types depending on the key parameters transmission probability, number of contacts per day, duration of the infectious period, different levels of clustering and varying proportions of repetitive contacts. RESULTS: The simulation runs under different parameter constellations provide the following results: The difference between both model types is highest for low numbers of contacts per day and low transmission probabilities. The number of contacts and the transmission probability have a higher influence on this difference than the duration of the infectious period. Even when only minor parts of the daily contacts are repetitive and clustered can there be relevant differences compared to a purely random mixing model. CONCLUSION: We show that random mixing models provide acceptable estimates of the total outbreak size if the number of contacts per day is high or if the per-contact transmission probability is high, as seen in typical childhood diseases such as measles. In the case of very short infectious periods, for instance, as in Norovirus, models assuming repeating contacts will also behave similarly as random mixing models. If the number of daily contacts or the transmission probability is low, as assumed for MRSA or Ebola, particular consideration should be given to the actual structure of potentially contagious contacts when designing the model. PMID- 19563625 TI - Participant observation of time allocation, direct patient contact and simultaneous activities in hospital physicians. AB - BACKGROUND: Hospital physicians' time is a critical resource in medical care. Two aspects are of interest. First, the time spent in direct patient contact - a key principle of effective medical care. Second, simultaneous task performance ('multitasking') which may contribute to medical error, impaired safety behaviour, and stress. There is a call for instruments to assess these aspects. A preliminary study to gain insight into activity patterns, time allocation and simultaneous activities of hospital physicians was carried out. Therefore an observation instrument for time-motion-studies in hospital settings was developed and tested. METHODS: 35 participant observations of internists and surgeons of a German municipal 300-bed hospital were conducted. Complete day shifts of hospital physicians on wards, emergency ward, intensive care unit, and operating room were continuously observed. Assessed variables of interest were time allocation, share of direct patient contact, and simultaneous activities. Inter-rater agreement of Kappa = .71 points to good reliability of the instrument. RESULTS: Hospital physicians spent 25.5% of their time at work in direct contact with patients. Most time was allocated to documentation and conversation with colleagues and nursing staff. Physicians performed parallel simultaneous activities for 17-20% of their work time. Communication with patients, documentation, and conversation with colleagues and nursing staff were the most frequently observed simultaneous activities. Applying logit-linear analyses, specific primary activities increase the probability of particular simultaneous activities. CONCLUSION: Patient related working time in hospitals is limited. The potential detrimental effects of frequently observed simultaneous activities on performance outcomes need further consideration. PMID- 19563626 TI - Essential health information available for India in the public domain on the internet. AB - BACKGROUND: Health information and statistics are important for planning, monitoring and improvement of the health of populations. However, the availability of health information in developing countries is often inadequate. This paper reviews the essential health information available readily in the public domain on the internet for India in order to broadly assess its adequacy and inform further development. METHODS: The essential sources of health-related information for India were reviewed. An extensive search of relevant websites and the PubMed literature database was conducted to identify the sources. For each essential source the periodicity of the data collection, the information it generates, the geographical level at which information is reported, and its availability in the public domain on the internet were assessed. RESULTS: The available information related to non-communicable diseases and injuries was poor. This is a significant gap as India is undergoing an epidemiological transition with these diseases/conditions accounting for a major proportion of disease burden. Information on infrastructure and human resources was primarily available for the public health sector, with almost none for the private sector which provides a large proportion of the health services in India. Majority of the information was available at the state level with almost negligible at the district level, which is a limitation for the practical implementation of health programmes at the district level under the proposed decentralisation of health services in India. CONCLUSION: This broad review of the essential health information readily available in the public domain on the internet for India highlights that the significant gaps related to non-communicable diseases and injuries, private health sector and district level information need to be addressed to further develop an effective health information system in India. PMID- 19563627 TI - Provision of palliative care for chronic heart failure inpatients: how much do we need? AB - BACKGROUND: Clinical guidance recommends early CHF palliative care intervention, but the magnitude of need is unknown and evidence-based referral criteria absent.This study aimed to: 1) Measure point prevalence of inpatients appropriate for palliative care. 2) Identify patient characteristics associated with palliative care appropriateness. 3) Propose evidence-based clinical referral criteria. METHODS: Census: all adult medical inpatient files in a UK tertiary teaching hospital were reviewed, identifying patients with CHF as a reason for current admission, using NYHA stage 3/4 classification, cross referenced with existing ECHO data. Each CHF patient was classified according to appropriateness for palliative care against a definition of unresolved pain and/or symptoms and/or psychosocial problems 7 days post admission. RESULTS: Three hundred and sixty-five patient files were reviewed, and 28 clinically identified as having CHF. Of these, 11 had confirmed unpreserved ejection fraction,16 of the 28 patients were appropriate for palliative care. Of the total inpatient population reviewed, 10 (2.7%) had both confirmed ejection fraction 45% also require palliative care. Our conservative criteria suggest a point prevalence of 2.7% of patients having both ejection fraction 70% and >80% in the GP202 and IPA220 gastric cancer cell lines, respectively. Integration of array CGH and microarray expression data provided a list of 134 and 50 candidate genes inactivated by nonsense mutation and deletion for GP202 and IPA220, respectively. We selected 12 candidate genes for mutation analysis. Of these, sequence analysis was performed on 11 genes. One gene, PLA2G4A, showed a silent mutation, and in two genes, CTSA and PTPRJ, missense mutations were detected. No nonsense mutations were detected in any of the 11 genes tested. CONCLUSION: Although UPF1 was substantially repressed, thus resulting in the inhibition of the NMD system, we did not find genes inactivated by nonsense mutations. Our results show that the GINI strategy leads to a high number of false positives. PMID- 19563645 TI - The association of XRCC1 gene single nucleotide polymorphisms with response to neoadjuvant chemotherapy in locally advanced cervical carcinoma. AB - BACKGROUND: Platinum-based neoadjuvant chemotherapy (NAC) is new therapeutic strategy for locally advanced cervical carcinoma, but the variables used to predict NAC response are still infrequently reported. The aim of our study was to investigate the association between XRCC1 gene single nucleotide polymorphisms (SNPs) and NAC response. METHODS: Seventy patients with locally advanced cervical carcinoma who underwent NAC were collected. SNPs of XRCC1 (at codon 194 and 399) and XRCC1 protein expression were detected. The association of XRCC1 gene SNPs and protein expression with NAC response were analyzed. RESULTS: Response to NAC was not statistically significant in three genotypes, Arg/Arg, Arg/Trp, Trp/Trp of XRCC1 at codon 194(X(2) = 1.243, P = 0.07), while responses were significantly different in genotypes Arg/Arg, Arg/Gln, Gln/Gln of XRCC1 at codon 399 (X(2) = 2.283, P = 0.020). The risk of failure to chemotherapy in the patients with a Gln allele(Arg/Gln+Gln/Gln) was significantly greater than that with Arg/Arg(OR = 3.254, 95%CI 1.708 approximately 14.951). The expression level of XRCC1 protein was significantly associated with response to NAC. Moreover, the genotype with the Gln allele(Arg/Gln+Gln/Gln) at codon 399, but not codon at 194, presented a significantly higher level of XRCC1 protein expression than that with Arg/Arg genotype (F = 2.699, p = 0.009). CONCLUSION: SNP of XRCC1 gene at codon 399 influences the response of cervical carcinoma to platinum-based NAC. This is probably due to changes in expression of XRCC1 protein, affecting response to chemotherapy. PMID- 19563646 TI - A simple method for co-segregation analysis to evaluate the pathogenicity of unclassified variants; BRCA1 and BRCA2 as an example. AB - BACKGROUND: Assessment of the clinical significance of unclassified variants (UVs) identified in BRCA1 and BRCA2 is very important for genetic counselling. The analysis of co-segregation of the variant with the disease in families is a powerful tool for the classification of these variants. Statistical methods have been described in literature but these methods are not always easy to apply in a diagnostic setting. METHODS: We have developed an easy to use method which calculates the likelihood ratio (LR) of an UV being deleterious, with penetrance as a function of age of onset, thereby avoiding the use of liability classes. The application of this algorithm is publicly available http://www.msbi.nl/cosegregation. It can easily be used in a diagnostic setting since it requires only information on gender, genotype, present age and/or age of onset for breast and/or ovarian cancer. RESULTS: We have used the algorithm to calculate the likelihood ratio in favour of causality for 3 UVs in BRCA1 (p.M18T, p.S1655F and p.R1699Q) and 5 in BRCA2 (p.E462G p.Y2660D, p.R2784Q, p.R3052W and p.R3052Q). Likelihood ratios varied from 0.097 (BRCA2, p.E462G) to 230.69 (BRCA2, p.Y2660D). Typing distantly related individuals with extreme phenotypes (i.e. very early onset cancer or old healthy individuals) are most informative and give the strongest likelihood ratios for or against causality. CONCLUSION: Although co segregation analysis on itself is in most cases insufficient to prove pathogenicity of an UV, this method simplifies the use of co-segregation as one of the key features in a multifactorial approach considerably. PMID- 19563647 TI - High levels of multidrug resistant tuberculosis in new and treatment-failure patients from the Revised National Tuberculosis Control Programme in an urban metropolis (Mumbai) in Western India. AB - BACKGROUND: India, China and Russia account for more than 62% of multidrug resistant tuberculosis (MDRTB) globally. Within India, locations like urban metropolitan Mumbai with its burgeoning population and high incidence of TB are suspected to be a focus for MDRTB. However apart from sporadic surveys at watched sites in the country, there has been no systematic attempt by the Revised National Tuberculosis Control Programme (RNTCP) of India to determine the extent of MDRTB in Mumbai that could feed into national estimates. Drug susceptibility testing (DST) is not routinely performed as a part of programme policy and public health laboratory infrastructure, is limited and poorly equipped to cope with large scale testing. METHODS: From April 2004 to January 2007 we determined the extent of drug resistance in 724 {493 newly diagnosed, previously untreated and 231 first line treatment failures (sputum-smear positive at the fifth month after commencement of therapy)} cases of pulmonary tuberculosis drawn from the RNTCP in four suboptimally performing municipal wards of Mumbai. The observations were obtained using a modified radiorespirometric Buddemeyer assay and validated by the Swedish Institute for Infectious Disease Control, Stockholm, a supranational reference laboratory. Data was analyzed utilizing SPSS 10.0 and Epi Info 2002. RESULTS: This study undertaken for the first time in RNTCP outpatients in Mumbai reveals a high proportion of MDRTB strains in both previously untreated (24%) and treatment-failure cases (41%). Amongst new cases, resistance to 3 or 4 drug combinations (amplified drug resistance) including isoniazid (H) and rifampicin (R), was greater (20%) than resistance to H and R alone (4%) at any point in time during the study. The trend for monoresistance was similar in both groups remaining highest to H and lowest to R. External quality control revealed good agreement for H and R resistance (k = 0.77 and 0.76 respectively). CONCLUSION: Levels of MDRTB are much higher in both previously untreated and first line treatment-failure cases in the selected wards in Mumbai than those projected by national estimates. The finding of amplified drug resistance suggests the presence of a well entrenched MDRTB scenario. This study suggests that a wider set of surveillance sites are needed to obtain a more realistic view of the true MDRTB rates throughout the country. This would assist in the planning of an adequate response to the diagnosis and care of MDRTB. PMID- 19563648 TI - N-acetylcysteine does not prevent contrast-induced nephropathy after cardiac catheterization in patients with diabetes mellitus and chronic kidney disease: a randomized clinical trial. AB - BACKGROUND: Patients with diabetes mellitus (DM) and chronic kidney disease (CKD) constitute to be a high-risk population for the development of contrast-induced nephropathy (CIN), in which the incidence of CIN is estimated to be as high as 50%. We performed this trial to assess the efficacy of N-acetylcysteine (NAC) in the prevention of this complication. METHODS: In a prospective, double-blind, placebo controlled, randomized clinical trial, we studied 90 patients undergoing elective diagnostic coronary angiography with DM and CKD (serum creatinine > or = 1.5 mg/dL for men and > or = 1.4 mg/dL for women). The patients were randomly assigned to receive either oral NAC (600 mg BID, starting 24 h before the procedure) or placebo, in adjunct to hydration. Serum creatinine was measured prior to and 48 h after coronary angiography. The primary end-point was the occurrence of CIN, defined as an increase in serum creatinine > or = 0.5 mg/dL (44.2 micromol/L) or > or = 25% above baseline at 48 h after exposure to contrast medium. RESULTS: Complete data on the outcomes were available on 87 patients, 45 of whom had received NAC. There were no significant differences between the NAC and placebo groups in baseline characteristics, amount of hydration, or type and volume of contrast used, except in gender (male/female, 20/25 and 34/11, respectively; P = 0.005) and the use of statins (62.2% and 37.8%, respectively; P = 0.034). CIN occurred in 5 out of 45 (11.1%) patients in the NAC group and 6 out of 42 (14.3%) patients in the placebo group (P = 0.656). CONCLUSION: There was no detectable benefit for the prophylactic administration of oral NAC over an aggressive hydration protocol in patients with DM and CKD. TRIAL REGISTRATION: NCT00808795. PMID- 19563649 TI - SPEM dysfunction and general schizotypy as measured by the SSQ: a controlled study. AB - BACKGROUND: SPEM dysfunction is a well-known phenomenon in schizophrenia. The principal aim of the present study was to examine whether SPEM dysfunction is already observable in subjects scoring high on a specific measure of schizotypy (SSQ General Schizotypy) that was selected because of its intimate relationship with schizophrenic prodromal unfolding. METHODS: Applying ANOVAs, we determined the relationship of subjects' scores on SSQ General Schizotypy and eye movements elicited by targets of different speed. We also examined whether there exists an association between our schizotypy measure and pupil size. RESULTS: We found more SPEM dysfunction in subjects scoring high on SSQ General Schizotypy than in subjects scoring average on that factor, irrespective of the speed of the target. No relationship was found between baseline pupil size and General Schizotypy. CONCLUSION: The present study provides additional evidence that SPEM dysfunction is associated with schizotypic features that precede the onset of schizophrenia and is already observable in general population subjects that show these features. PMID- 19563650 TI - Seasonal variation in objectively assessed physical activity among children and adolescents in Norway: a cross-sectional study. AB - BACKGROUND: The literature on seasonality in children and youth's physical activity participation is inconsistent. The aims of this study were to: 1) compare physical activity across seasons and describe activity patterns within seasons, and 2) to determine compliance with current physical activity recommendations across seasons among 9- and 15-year-olds living in a climatically diverse country. METHODS: Participants were 2,299 9- and 15-year-olds from all regions in Norway. Physical activity was assessed using the Actigraph accelerometer for 4 consecutive days. Physical activity data were collected during winter, spring and fall. General linear models were used to study the associations between physical activity and season. RESULTS: Nine-year-old children had significantly higher mean physical activity levels in spring than in winter and fall. In the two latter seasons, physical activity levels were especially low after school hours and on weekends. Logistic regression models demonstrated that 9-year-olds had 3.3 times (95% Confidence Interval (CI): 2.08, 5.18) higher odds of meeting recommended levels of physical activity in spring than in winter. No associations were found between mean physical activity level and season among the 15-year-olds. However, the adolescents also had higher odds (OR = 1.56; 95% CI: 1.05, 2.32) of meeting the physical activity recommendations in spring than in winter. CONCLUSION: In a large population-based sample, we observed substantial seasonal differences in physical activity among 9-year-olds, and the activity pattern varied across the seasons. The results emphasize the need to take season into account when developing physical activity interventions for children. Season appears to have less influence on adolescent's physical activity; interventions for increasing physical activity in this group could therefore be implemented throughout the year. PMID- 19563652 TI - Habitat adaptation rather than genetic distance correlates with female preference in fire salamanders (Salamandra salamandra). AB - BACKGROUND: Although some mechanisms of habitat adaptation of conspecific populations have been recently elucidated, the evolution of female preference has rarely been addressed as a force driving habitat adaptation in natural settings. Habitat adaptation of fire salamanders (Salamandra salamandra), as found in Middle Europe (Germany), can be framed in an explicit phylogeographic framework that allows for the evolution of habitat adaptation between distinct populations to be traced. Typically, females of S. salamandra only deposit their larvae in small permanent streams. However, some populations of the western post-glacial recolonization lineage use small temporary ponds as larval habitats. Pond larvae display several habitat-specific adaptations that are absent in stream-adapted larvae. We conducted mate preference tests with females from three distinct German populations in order to determine the influence of habitat adaptation versus neutral genetic distance on female mate choice. Two populations that we tested belong to the western post-glacial recolonization group, but are adapted to either stream or pond habitats. The third population is adapted to streams but represents the eastern recolonization lineage. RESULTS: Despite large genetic distances with FST values around 0.5, the stream-adapted females preferred males from the same habitat type regardless of genetic distance. Conversely, pond adapted females did not prefer males from their own population when compared to stream-adapted individuals of either lineage. CONCLUSION: A comparative analysis of our data showed that habitat adaptation rather than neutral genetic distance correlates with female preference in these salamanders, and that habitat dependent female preference of a specific pond-reproducing population may have been lost during adaptation to the novel environmental conditions of ponds. PMID- 19563651 TI - Effect of questionnaire structure on recall of drug utilization in a population of university students. AB - BACKGROUND: Self-reported data are a common source of information about drug exposure. Modes of data collection differ considerably and the questionnaire's structure may affect prevalence estimates. We compared the recall of medication use evaluated by means of two questionnaires differing in structure and length. METHODS: Drug utilization was assessed by two alternative versions of a questionnaire (A - 4 pages, including specific questions for 12 indications/pharmacological groups and one question for "other medicines"; B - 1 page, including 1 open-ended question to cover overall drug consumption). Each of 32 classes in a private University in Maputo, Mozambique, was randomly assigned questionnaire A (233 participants) or B (276 participants). Logistic regression (allowing for clustering by classroom) was used to compare the two groups in terms of socio-demographic characteristics and medication used during the previous month. RESULTS: Overall, 67.4% of the subjects had used at least one drug during the previous month. The following prevalences were greater among participants completing questionnaire A: use of drugs from two or more pharmacological groups (60.5% vs. 34.4%, p < 0.001), use of two or more drugs (66.2% vs. 43.0%, p < 0.001), and use of antibiotics (14.6% vs. 6.9%, p = 0.001), antifungals (9.4% vs. 4.0%, p = 0.013), antiparasitics (5.6% vs. 1.8%, p = 0.031) and antacids (8.6% vs. 3.6%, p = 0.024). Information about duration of treatment and medical advice was more complete with version A. CONCLUSION: The indication/drug-specific questions (questionnaire A) revealed a significantly higher prevalence of use of medicines - antibiotics, antifungals, antiparasitics and antacids - without compromising the completeness of the information. PMID- 19563653 TI - Ontology-oriented retrieval of putative microRNAs in Vitis vinifera via GrapeMiRNA: a web database of de novo predicted grape microRNAs. AB - BACKGROUND: Two complete genome sequences are available for Vitis vinifera Pinot noir. Based on the sequence and gene predictions produced by the IASMA, we performed an in silico detection of putative microRNA genes and of their targets, and collected the most reliable microRNA predictions in a web database. The application is available at http://www.itb.cnr.it/ptp/grapemirna/. DESCRIPTION: The program FindMiRNA was used to detect putative microRNA genes in the grape genome. A very high number of predictions was retrieved, calling for validation. Nine parameters were calculated and, based on the grape microRNAs dataset available at miRBase, thresholds were defined and applied to FindMiRNA predictions having targets in gene exons. In the resulting subset, predictions were ranked according to precursor positions and sequence similarity, and to target identity. To further validate FindMiRNA predictions, comparisons to the Arabidopsis genome, to the grape Genoscope genome, and to the grape EST collection were performed. Results were stored in a MySQL database and a web interface was prepared to query the database and retrieve predictions of interest. CONCLUSION: The GrapeMiRNA database encompasses 5,778 microRNA predictions spanning the whole grape genome. Predictions are integrated with information that can be of use in selection procedures. Tools added in the web interface also allow to inspect predictions according to gene ontology classes and metabolic pathways of targets. The GrapeMiRNA database can be of help in selecting candidate microRNA genes to be validated. PMID- 19563654 TI - NLStradamus: a simple Hidden Markov Model for nuclear localization signal prediction. AB - BACKGROUND: Nuclear localization signals (NLSs) are stretches of residues within a protein that are important for the regulated nuclear import of the protein. Of the many import pathways that exist in yeast, the best characterized is termed the 'classical' NLS pathway. The classical NLS contains specific patterns of basic residues and computational methods have been designed to predict the location of these motifs on proteins. The consensus sequences, or patterns, for the other import pathways are less well-understood. RESULTS: In this paper, we present an analysis of characterized NLSs in yeast, and find, despite the large number of nuclear import pathways, that NLSs seem to show similar patterns of amino acid residues. We test current prediction methods and observe a low true positive rate. We therefore suggest an approach using hidden Markov models (HMMs) to predict novel NLSs in proteins. We show that our method is able to consistently find 37% of the NLSs with a low false positive rate and that our method retains its true positive rate outside of the yeast data set used for the training parameters. CONCLUSION: Our implementation of this model, NLStradamus, is made available at: (http://www.moseslab.csb.utoronto.ca/NLStradamus/). PMID- 19563655 TI - An evaluation of the performance in the UK Royal College of Anaesthetists primary examination by UK medical school and gender. AB - BACKGROUND: There has been comparatively little consideration of the impact that the changes to undergraduate curricula might have on postgraduate academic performance. This study compares the performance of graduates by UK medical school and gender in the Multiple Choice Question (MCQ) section of the first part of the Fellowship of the Royal College of Anaesthetists (FRCA) examination. METHODS: Data from each sitting of the MCQ section of the primary FRCA examination from June 1999 to May 2008 were analysed for performance by medical school and gender. RESULTS: There were 4983 attempts at the MCQ part of the examination by 3303 graduates from the 19 United Kingdom medical schools. Using the standardised overall mark minus the pass mark graduates from five medical schools performed significantly better than the mean for the group and five schools performed significantly worse than the mean for the group. Males performed significantly better than females in all aspects of the MCQ - physiology, mean difference = 3.0% (95% CI 2.3, 3.7), p < 0.001; pharmacology, mean difference = 1.7% (95% CI 1.0, 2.3), p < 0.001; physics with clinical measurement, mean difference = 3.5% (95% CI 2.8, 4.1), p < 0.001; overall mark, mean difference = 2.7% (95% CI 2.1, 3.3), p < 0.001; and standardised overall mark minus the pass mark, mean difference = 2.5% (95% CI 1.9, 3.1), p < 0.001. Graduates from three medical schools that have undergone the change from Traditional to Problem Based Learning curricula did not show any change in performance in any aspects of the MCQ pre and post curriculum change. CONCLUSION: Graduates from each of the medical schools in the UK do show differences in performance in the MCQ section of the primary FRCA, but significant curriculum change does not lead to deterioration in post graduate examination performance. Whilst females now outnumber males taking the MCQ, they are not performing as well as the males. PMID- 19563656 TI - Integrated analysis of DNA copy number and gene expression microarray data using gene sets. AB - BACKGROUND: Genes that play an important role in tumorigenesis are expected to show association between DNA copy number and RNA expression. Optimal power to find such associations can only be achieved if analysing copy number and gene expression jointly. Furthermore, some copy number changes extend over larger chromosomal regions affecting the expression levels of multiple resident genes. RESULTS: We propose to analyse copy number and expression array data using gene sets, rather than individual genes. The proposed model is robust and sensitive. We re-analysed two publicly available datasets as illustration. These two independent breast cancer datasets yielded similar patterns of association between gene dosage and gene expression levels, in spite of different platforms having been used. Our comparisons show a clear advantage to using sets of genes' expressions to detect associations with long-spanning, low-amplitude copy number aberrations. In addition, our model allows for using additional explanatory variables and does not require mapping between copy number and expression probes. CONCLUSION: We developed a general and flexible tool for integration of multiple microarray data sets, and showed how the identification of genes whose expression is affected by copy number aberrations provides a powerful approach to prioritize putative targets for functional validation. PMID- 19563657 TI - The pluripotency factor LIN28 marks undifferentiated spermatogonia in mouse. AB - BACKGROUND: Life-long production of spermatozoa depends on spermatogonial stem cells. Spermatogonial stem cells exist among the most primitive population of germ cells - undifferentiated spermatogonia. Transplantation experiments have demonstrated the functional heterogeneity of undifferentiated spermatogonia. Although the undifferentiated spermatogonia can be topographically divided into As (single), Apr (paired), and Aal (aligned) spermatogonia, subdivision of this primitive cell population using cytological markers would greatly facilitate characterization of their functions. RESULTS: In the present study, we show that LIN28, a pluripotency factor, is specifically expressed in undifferentiated spermatogonia (As, Apr, and Aal) in mouse. Ngn3 also specifically labels undifferentiated spermatogonia. We used Ngn3-GFP knockin mice, in which GFP expression is under the control of all Ngn3 transcription regulatory elements. Remarkably, Ngn3-GFP is only expressed in approximately 40% of LIN28-positive As (single) cells. The percentage of Ngn3-GFP-positive clusters increases dramatically with the chain length of interconnected spermatogonia. CONCLUSION: Our study demonstrates that LIN28 specifically marks undifferentiated spermatogonia in mice. These data, together with previous studies, suggest that the LIN28-expressing undifferentiated spermatogonia exist as two subpopulations: Ngn3-GFP-negative (high stem cell potential) and Ngn3-GFP-positive (high differentiation commitment). Furthermore, Ngn3-GFP-negative cells are found in chains of Ngn3-GFP-positive spermatogonia, suggesting that cells in the Aal spermatogonia could revert to a more primitive state. PMID- 19563658 TI - Molecular characterization of EGFR, PDGFRA and VEGFR2 in cervical adenosquamous carcinoma. AB - BACKGROUND: Adenosquamous carcinoma of the uterine cervix is an infrequent but aggressive subtype of cervical cancer. A better understanding of its biological behaviour is warranted to define more accurate prognosis and therapeutic targets. Currently, the blockage of receptor tyrosine kinase (RTKs) activity is an efficient therapeutic strategy for many different cancers. The objective of this study was to investigate EGFR, PDGFRA and VEGFR2 RTKs overexpression and activating gene mutations in a cohort of 30 adenosquamous carcinomas of the uterine cervix. METHODS: EGFR, PDGFRA and VEGFR2 immunohistochemistry was performed in all samples, followed by DNA isolation from the gross macroscopically dissection of the neoplastic area. Screening for EGFR (exons 18 21) and PDGFRA (exons 12, 14 and 18) mutations was done by PCR--single-strand conformational polymorphism (PCR-SSCP). RESULTS: Despite the presence of EGFR immunohistochemical positive reactions in 43% (13/30) of the samples, no EGFR activating mutations in the hotspot region (exons 18-21) were identified. A silent base substitution (CAG>CAA) in EGFR exon 20 at codon 787 (Q787Q) was found in 17 cases (56%). All PDGFRA immunohistochemical reactions were positive and consistently observed in the stromal component, staining fibroblasts and endothelial cells, as well as in the cytoplasm of malignant cells. No activating PDGFRA mutations were found, yet, several silent mutations were observed, such as a base substitution in exon 12 (CCA>CCG) at codon 567 (P567P) in 9 cases and in exon 18 (GTC>GTT) at codon 824 (V824V) in 4 cases. We also observed the presence of base substitutions in intron 14 (IVS14+3G>A and IVS14+49G>A) in two different cases, and in intron 18 (IVS18-50insA) in 4 cases. VEGFR2 positivity was observed in 22 of 30 cases (73.3%), and was significantly associated with lack of metastasis (p=0.038). CONCLUSION: This is the most extensive analysis of EGFR, PDGFRA and VEGFR2 in cervical adenosquamous carcinomas. Despite the absence of EGFR and PDGFRA activating mutations, the presence of overexpression of these three important therapeutic targets in a subset of cases may be important in predicting the sensitivity of adenosquamous carcinoma to specific anti-RTKs drugs. PMID- 19563660 TI - Airborne particulate matter PM2.5 from Mexico City affects the generation of reactive oxygen species by blood neutrophils from asthmatics: an in vitro approach. AB - BACKGROUND: The Mexico City Metropolitan Area is densely populated, and toxic air pollutants are generated and concentrated at a higher rate because of its geographic characteristics. It is well known that exposure to particulate matter, especially to fine and ultra-fine particles, enhances the risk of cardio respiratory diseases, especially in populations susceptible to oxidative stress. The aim of this study was to evaluate the effect of fine particles on the respiratory burst of circulating neutrophils from asthmatic patients living in Mexico City. METHODS: In total, 6 subjects diagnosed with mild asthma and 11 healthy volunteers were asked to participate. Neutrophils were isolated from peripheral venous blood and incubated with fine particles, and the generation of reactive oxygen species was recorded by chemiluminescence. We also measured plasma lipoperoxidation susceptibility and plasma myeloperoxidase and paraoxonase activities by spectrophotometry. RESULTS: Asthmatic patients showed significantly lower plasma paraoxonase activity, higher susceptibility to plasma lipoperoxidation and an increase in myeloperoxidase activity that differed significantly from the control group. In the presence of fine particles, neutrophils from asthmatic patients showed an increased tendency to generate reactive oxygen species after stimulation with fine particles (PM2.5). CONCLUSION: These findings suggest that asthmatic patients have higher oxidation of plasmatic lipids due to reduced antioxidant defense. Furthermore, fine particles tended to increase the respiratory burst of blood human neutrophils from the asthmatic group.On the whole, increased myeloperoxidase activity and susceptibility to lipoperoxidation with a concomitant decrease in paraoxonase activity in asthmatic patients could favor lung infection and hence disrupt the control of asthmatic crises. PMID- 19563659 TI - Outpatient alcoholism treatment--24-month outcome and predictors of outcome. AB - OBJECTIVES: To study the value of demographic and alcohol-related variables for predicting 24-month treatment outcome in an outpatient setting. METHODS: Prospective observational study with 92 alcohol-dependent patients. Assessments were made by personal interviews at the beginning and end of therapy, and at the 24-month follow-up. Univariate and logistic regression analyses were performed. RESULTS: The mean age was 46.0 (SD = 9.9) years. There were 58 males (65.2%) and 31 females (34.8%). Of the 67 patients interviewed at 2-year follow-up, 58% were abstinent and 79% improved. Differences between abstainers and non-abstainers were found for number of previous detoxifications, and number of patients attempted suicides. In addition, female gender and a higher number of prior treatments predicted negative treatment outcome. CONCLUSION: Matching patients to different types of treatment by means of empirically based characteristics may help to improve outcome but research has failed to establish reliable predictors in that area. Data from this follow-up study confirm the role of certain clinical outcome predictors. Additionally, results give further evidence for outpatient treatment as an effective setting for alcohol-dependent patients as indicated by a favourable retention rate (84%) and outcome (minimum abstinence rate 44%). PMID- 19563661 TI - Structure-activity relationship analysis of cytotoxic cyanoguanidines: selection of CHS 828 as candidate drug. AB - BACKGROUND: N-(6-(4-chlorophenoxy)hexyl)-N'-cyano-N''-4-pyridyl guanidine) (CHS 828) is the first candidate drug from a novel group of anti-tumour agents - the pyridyl cyanoguanidines, shown to be potent compounds interfering with cellular metabolism (inhibition of nicotinamide phosphoribosyl transferase) and NF-kappaB signalling. Substituted cyanoguanidines are also found in anti-hypertensive agents such as the potassium channel opener pinacidil (N-cyano-N'-(4-pyridyl)-N'' (1,2,2-trimethylpropyl)guanidine) and histamine-II receptor antagonists (e.g. cimetidine, N-cyano-N'-methyl-N''-[2-[[(5-methylimidazol-4 yl]methyl]thio]ethyl)guanidine). In animal studies, CHS 828 has shown very promising activity, and phase I and II studies resulted in further development of a with a water soluble prodrug. FINDINGS: To study the structural requirements for cyanoguanidine cytotoxicity a set of 19 analogues were synthesized. The cytotoxic effects were then studied in ten cell lines selected for different origins and mechanisms of resistance, using the fluorometric microculture cytotoxicity assay (FMCA). The compounds showed varying cytotoxic activity even though the dose-response curves for some analogues were very shallow. Pinacidil and cimetidine were found to be non-toxic in all ten cell lines. Starting with cyanoguanidine as the crucial core it was shown that 4-pyridyl substitution was more efficient than was 3-pyridyl substitution. The 4-pyridyl cyanoguanidine moiety should be linked by an alkyl chain, optimally a hexyl, heptyl or octyl chain, to a bulky end group. The exact composition of this end group did not seem to be of crucial importance; when the end group was a mono-substituted phenyl ring it was shown that the preferred position was 4-substitution, followed by 3- and, finally, 2-substitution as the least active. Whether the substituent was a chloro, nitro or methoxy substituent seemed to be of minor importance. Finally, the activity patterns in the ten cell lines were compared. Substances with similar structures correlated well, whilst substances with large differences in molecular structure demonstrated lower correlation coefficients. CONCLUSION: According to this structure-activity relationship (SAR) study, CHS 828 meets the requirements for optimal cytotoxic activity for this class of compounds. PMID- 19563662 TI - Novel sequential ChIP and simplified basic ChIP protocols for promoter co occupancy and target gene identification in human embryonic stem cells. AB - BACKGROUND: The investigation of molecular mechanisms underlying transcriptional regulation, particularly in embryonic stem cells, has received increasing attention and involves the systematic identification of target genes and the analysis of promoter co-occupancy. High-throughput approaches based on chromatin immunoprecipitation (ChIP) have been widely used for this purpose. However, these approaches remain time-consuming, expensive, labor-intensive, involve multiple steps, and require complex statistical analysis. Advances in this field will greatly benefit from the development and use of simple, fast, sensitive and straightforward ChIP assay and analysis methodologies. RESULTS: We initially developed a simplified, basic ChIP protocol that combines simplicity, speed and sensitivity. ChIP analysis by real-time PCR was compared to analysis by densitometry with the ImageJ software. This protocol allowed the rapid identification of known target genes for SOX2, NANOG, OCT3/4, SOX17, KLF4, RUNX2, OLIG2, SMAD2/3, BMI-1, and c-MYC in a human embryonic stem cell line. We then developed a novel Sequential ChIP protocol to investigate in vivo promoter co occupancy, which is basically characterized by the absence of antibody-antigen disruption during the assay. It combines centrifugation of agarose beads and magnetic separation. Using this Sequential ChIP protocol we found that c-MYC associates with the SOX2/NANOG/OCT3/4 complex and identified a novel RUNX2/BMI 1/SMAD2/3 complex in BG01V cells. These two TF complexes associate with two distinct sets of target genes. The RUNX2/BMI-1/SMAD2/3 complex is associated predominantly with genes not expressed in undifferentiated BG01V cells, consistent with the reported role of those TFs as transcriptional repressors. CONCLUSION: These simplified basic ChIP and novel Sequential ChIP protocols were successfully tested with a variety of antibodies with human embryonic stem cells, generated a number of novel observations for future studies and might be useful for high-throughput ChIP-based assays. PMID- 19563664 TI - Salter-Harris II injury of the proximal tibial epiphysis with both vascular compromise and compartment syndrome: a case report. AB - We present a case of a Salter-Harris II injury to the proximal tibia associated with both vascular compromise and compartment syndrome. The potential complications of this injury are limb threatening and the neurovasular status of the limb should be continually monitored. Maintaining anatomic reduction is difficult and fixation may be needed to achieve optimal results. PMID- 19563663 TI - Phenotype-dependent apoptosis signalling in mesothelioma cells after selenite exposure. AB - BACKGROUND: Selenite is a promising anticancer agent which has been shown to induce apoptosis in malignant mesothelioma cells in a phenotype-dependent manner, where cells of the chemoresistant sarcomatoid phenotype are more sensitive. METHODS: In this paper, we investigate the apoptosis signalling mechanisms in sarcomatoid and epithelioid mesothelioma cells after selenite treatment. Apoptosis was measured with the Annexin-PI assay. The mitochondrial membrane potential, the expression of Bax, Bcl-XL, and the activation of caspase-3 were assayed with flow cytometry and a cytokeratin 18 cleavage assay. Signalling through JNK, p38, p53, and cathepsins B, D, and E was investigated with chemical inhibitors. Furthermore, the expression, nuclear translocation and DNA-binding activity of p53 was investigated using ICC, EMSA and the monitoring of p21 expression as a downstream event. Levels of thioredoxin (Trx) were measured by ELISA. RESULTS: In both cell lines, 10 microM selenite caused apoptosis and a marked loss of mitochondrial membrane potential. Bax was up-regulated only in the sarcomatoid cell line, while the epithelioid cell line down-regulated Bcl-XL and showed greater caspase-3 activation. Nuclear translocation of p53 was seen in both cell lines, but very little p21 expression was induced. Chemical inhibition of p53 did not protect the cells from apoptosis. p53 lost its DNA binding ability after selenite treatment and was enriched in an inactive form. Levels of thioredoxin decreased after selenite treatment. Chemical inhibition of MAP kinases and cathepsins showed that p38 and cathepsin B had some mediatory effect while JNK had an anti-apoptotic role. CONCLUSION: We delineate pathways of apoptosis signalling in response to selenite, showing differences between epithelioid and sarcomatoid mesothelioma cells. These differences may partly explain why sarcomatoid cells are more sensitive to selenite. PMID- 19563665 TI - Predictors of anti-convulsant treatment failure in children presenting with malaria and prolonged seizures in Kampala, Uganda. AB - BACKGROUND: In endemic areas, falciparum malaria remains the leading cause of seizures in children presenting to emergency departments. In addition, seizures in malaria have been shown to increase morbidity and mortality in these patients. The management of seizures in malaria is sometimes complicated by the refractory nature of these seizures to readily available anti-convulsants. The objective of this study was to determine predictors of anti-convulsant treatment failure and seizure recurrence after initial control among children with malaria. METHODS: In a previous study, the efficacy and safety of buccal midazolam was compared to that of rectal diazepam in the treatment of prolonged seizures in children aged three months to 12 years in Kampala, Uganda. For this study, predictive models were used to determine risk factors for anti-convulsant treatment failure and seizure recurrence among the 221 of these children with malaria. RESULTS: Using predictive models, focal seizures (OR 3.21; 95% CI 1.42-7.25, p = 0.005), cerebral malaria (OR 2.43; 95% CI 1.20-4.91, p = 0.01) and a blood sugar >or=200 mg/dl at presentation (OR 2.84; 95% CI 1.11-7.20, p = 0.02) were independent predictors of treatment failure (seizure persistence beyond 10 minutes or recurrence within one hour of treatment). Predictors of seizure recurrence included: 1) cerebral malaria (HR 3.32; 95% CI 1.94-5.66, p < 0.001), 2) presenting with multiple seizures (HR 2.45; 95% CI 1.42-4.23, p = 0.001), 3) focal seizures (HR 2.86; 95% CI 1.49-5.49, p = 0.002), 4) recent use of diazepam (HR 2.43; 95% CI 1.19-4.95, p = 0.01) and 5) initial control of the seizure with diazepam (HR 1.96; 95% CI 1.16-3.33, p = 0.01). CONCLUSION: Specific predictors, including cerebral malaria, can identify patients with malaria at risk of anti convulsant treatment failure and seizure recurrence. PMID- 19563666 TI - CXCR4 expression on circulating pan-cytokeratin positive cells is associated with survival in patients with advanced non-small cell lung cancer. AB - BACKGROUND: The CXC chemokine, CXCL12, and its receptor, CXCR4 promote metastases of a variety of solid tumors, including non-small cell lung cancer (NSCLC). The expression of CXCR4 on tumor cells may represent a critical biomarker for their propensity to metastasize. This study was performed to evaluate the hypothesis that co-expression of pan-cytokeratin and CXCR4 may be a prognostic marker for patients with advanced NSCLC. METHODS: We evaluated CXCR4 levels on circulating pan-cytokeratin positive cells from patients with NSCLC. NSCLC tumor and metastases were also assessed for the presence of CXCR4. RESULTS: Pan-cytokeratin positive cells were increased in the circulation of patients with NSCLC, as compared to normal control subjects. Patients with pan-cytokeratin +/CXCR4+ = 2,500 cells/ml had a significant improvement in median survival when compared with patients with pan-cytokeratin +/CXCR4+ >2,500 cells/ml (not achieved versus 14 weeks). CXCR4 expression was found on NSCLC tumors and at sites of tumor metastasis. CONCLUSION: This study suggests that CXCR4 may be a prognostic marker in NSCLC, and provides hypothesis-generating results, which may be important in determining metastatic potential. In future studies, we will prospectively evaluate the prognostic significance of pan-cytokeratin/CXCR4+ cells, and determine the mechanisms involved in the regulation of CXCR4 expression on tumor cells in a larger patient population. PMID- 19563667 TI - Thoracic spine pain in the general population: prevalence, incidence and associated factors in children, adolescents and adults. A systematic review. AB - BACKGROUND: Thoracic spine pain (TSP) is experienced across the lifespan by healthy individuals and is a common presentation in primary healthcare clinical practice. However, the epidemiological characteristics of TSP are not well documented compared to neck and low back pain. A rigorous evaluation of the prevalence, incidence, correlates and risk factors needs to be undertaken in order for epidemiologic data to be meaningfully used to develop evidence-based prevention and treatment recommendations for TSP. METHODS: A systematic review method was followed to report the evidence describing prevalence, incidence, associated factors and risk factors for TSP among the general population. Nine electronic databases were systematically searched to identify studies that reported either prevalence, incidence, associated factors (cross-sectional study) or risk factors (prospective study) for TSP in healthy children, adolescents or adults. Studies were evaluated for level of evidence and method quality. RESULTS: Of the 1389 studies identified in the literature, 33 met the inclusion criteria for this systematic review. The mean (SD) quality score (out of 15) for the included studies was 10.5 (2.0). TSP prevalence data ranged from 4.0-72.0% (point), 0.5-51.4% (7-day), 1.4-34.8% (1-month), 4.8-7.0% (3-month), 3.5-34.8% (1 year) and 15.6-19.5% (lifetime). TSP prevalence varied according to the operational definition of TSP. Prevalence for any TSP ranged from 0.5-23.0%, 15.8 34.8%, 15.0-27.5% and 12.0-31.2% for 7-day, 1-month, 1-year and lifetime periods, respectively. TSP associated with backpack use varied from 6.0-72.0% and 22.9 51.4% for point and 7-day periods, respectively. TSP interfering with school or leisure ranged from 3.5-9.7% for 1-year prevalence. Generally, studies reported a higher prevalence for TSP in child and adolescent populations, and particularly for females. The 1 month, 6 month, 1 year and 25 year incidences were 0-0.9%, 10.3%, 3.8-35.3% and 9.8% respectively. TSP was significantly associated with: concurrent musculoskeletal pain; growth and physical; lifestyle and social; backpack; postural; psychological; and environmental factors. Risk factors identified for TSP in adolescents included age (being older) and poorer mental health. CONCLUSION: TSP is a common condition in the general population. While there is some evidence for biopsychosocial associations it is limited and further prospectively designed research is required to inform prevention and management strategies. PMID- 19563668 TI - Comparison of two label-free global quantitation methods, APEX and 2D gel electrophoresis, applied to the Shigella dysenteriae proteome. AB - The in vitro stationary phase proteome of the human pathogen Shigella dysenteriae serotype 1 (SD1) was quantitatively analyzed in Coomassie Blue G250 (CBB)-stained 2D gels. More than four hundred and fifty proteins, of which 271 were associated with distinct gel spots, were identified. In parallel, we employed 2D-LC-MS/MS followed by the label-free computationally modified spectral counting method APEX for absolute protein expression measurements. Of the 4502 genome-predicted SD1 proteins, 1148 proteins were identified with a false positive discovery rate of 5% and quantitated using 2D-LC-MS/MS and APEX. The dynamic range of the APEX method was approximately one order of magnitude higher than that of CBB-stained spot intensity quantitation. A squared Pearson correlation analysis revealed a reasonably good correlation (R2 = 0.67) for protein quantities surveyed by both methods. The correlation was decreased for protein subsets with specific physicochemical properties, such as low Mr values and high hydropathy scores. Stoichiometric ratios of subunits of protein complexes characterized in E. coli were compared with APEX quantitative ratios of orthologous SD1 protein complexes. A high correlation was observed for subunits of soluble cellular protein complexes in several cases, demonstrating versatile applications of the APEX method in quantitative proteomics. PMID- 19563669 TI - Targeting inhibitor of apoptosis proteins in combination with ErbB antagonists in breast cancer. AB - INTRODUCTION: Inhibitor of apoptosis (IAPs) proteins are a family of proteins that can block apoptosis in normal cells and have been suggested to cause resistance to apoptosis in cancer. Overexpression of oncogenic receptor tyrosine kinases is common in breast cancer; in particular 20% of all cases show elevated Her2. Despite clinical success with the use of targeted therapies, such as Trastuzumab, only up to 35% of Her2-positive patients initially respond. We reasoned that IAP-mediated apoptosis resistance might contribute to this insensitivity to receptor tyrosine kinase therapy, in particular ErbB antagonists. Here we examine the levels of IAPs in breast cancer and evaluate whether targeting IAPs can enhance apoptosis in response to growth factor receptor antagonists and TRAIL. METHODS: IAP levels were examined in a breast cancer cell line panel and in patient samples. IAPs were inhibited using siRNA or cell permeable mimetics of endogenous inhibitors. Cells were then exposed to TRAIL, Trastuzumab, Lapatinib, or Gefitinib for 48 hours. Examining nuclear morphology and staining for cleaved caspase 3 was used to score apoptosis. Proliferation was examined by Ki67 staining. RESULTS: Four members of the IAP family, Survivin, XIAP, cIAP1 and cIAP2, were all expressed to varying extents in breast cancer cell lines or tumours. MDAMB468, BT474 and BT20 cells all expressed XIAP to varying extents. Depleting the cells of XIAP overcame the intrinsic resistance of BT20 and MDAMB468 cells to TRAIL. Moreover, siRNA-based depletion of XIAP or use of a Smac mimetic to target multiple IAPs increased apoptosis in response to the ErbB antagonists, Trastuzumab, Lapatinib or Gefitinib in Her2 overexpressing BT474 cells, or Gefitinib in EGFR-overexpressing MDAMB468 cells. CONCLUSIONS: The novel findings of this study are that multiple IAPs are concomitantly expressed in breast cancers, and that, in combination with clinically relevant Her2 treatments, IAP antagonists promote apoptosis and reduce the cell turnover index of breast cancers. We also show that combination therapy of IAP antagonists with some pro-apoptotic agents (for example, TRAIL) enhances apoptosis of breast cancer cells. In some cases (for example, MDAMB468 cells), the enhanced apoptosis is profound. PMID- 19563670 TI - A membrane-bound matrix-metalloproteinase from Nicotiana tabacum cv. BY-2 is induced by bacterial pathogens. AB - BACKGROUND: Plant matrix metalloproteinases (MMP) are conserved proteolytic enzymes found in a wide range of monocotyledonous and dicotyledonous plant species. Acting on the plant extracellular matrix, they play crucial roles in many aspects of plant physiology including growth, development and the response to stresses such as pathogen attack. RESULTS: We have identified the first tobacco MMP, designated NtMMP1, and have isolated the corresponding cDNA sequence from the tobacco suspension cell line BY-2. The overall domain structure of NtMMP1 is similar to known MMP sequences, although certain features suggest it may be constitutively active rather than dependent on proteolytic processing. The protein appears to be expressed in two forms with different molecular masses, both of which are enzymatically active as determined by casein zymography. Exchanging the catalytic domain of NtMMP1 with green fluorescent protein (GFP) facilitated subcellular localization by confocal laser scanning microscopy, showing the protein is normally inserted into the plasma membrane. The NtMMP1 gene is expressed constitutively at a low level but can be induced by exposure to bacterial pathogens. CONCLUSION: Our biochemical analysis of NtMMP1 together with bioinformatic data on the primary sequence indicate that NtMMP1 is a constitutively-active protease. Given its induction in response to bacterial pathogens and its localization in the plasma membrane, we propose a role in pathogen defense at the cell periphery. PMID- 19563671 TI - Effects of cardiovascular lifestyle change on lipoprotein subclass profiles defined by nuclear magnetic resonance spectroscopy. AB - BACKGROUND: Low-density lipoprotein (LDL) cholesterol lowering is a primary goal in clinical management of patients with cardiovascular disease, but traditional cholesterol levels may not accurately reflect the true atherogenicity of plasma lipid profiles. The size and concentration of lipoprotein particles, which transport cholesterol and triglycerides, may provide additional information for accurately assessing cardiovascular risk. This study evaluated changes in plasma lipoprotein profiles determined by nuclear magnetic resonance (NMR) spectroscopy in patients participating in a prospective, nonrandomized lifestyle modification program designed to reverse or stabilize progression of coronary artery disease (CAD) to improve our understanding of lipoprotein management in cardiac patients. RESULTS: The lifestyle intervention was effective in producing significant changes in lipoprotein subclasses that contribute to CAD risk. There was a clear beneficial effect on the total number of LDL particles (-8.3%, p < 0.05 compared to matched controls), small dense LDL particles (-9.5%, p < 0.05), and LDL particle size (+0.8%; p < 0.05). Likewise, participants showed significant improvement in traditional CAD risk factors such as body mass index (-9.9%, p < 0.01 compared to controls), total cholesterol (-5.5%, p < 0.05), physical fitness (+37.2%, p < 0.01), and future risk for CAD (-7.9%, p < 0.01). Men and women responded differently to the program for all clinically-relevant variables, with men deriving greater benefit in terms of lipoprotein atherogenicity. Plasma lipid and lipoprotein responses to the lifestyle change program were not confounded by lipid-lowering medications. CONCLUSION: In at risk patients motivated to participate, an intensive lifestyle change program can effectively alter traditional CAD risk factors and plasma lipoprotein subclasses and may reduce risk for cardiovascular events. Improvements in lipoprotein subclasses are more evident in men compared to women. PMID- 19563672 TI - Myeloid dendritic cells correlate with clinical response whereas plasmacytoid dendritic cells impact autoantibody development in rheumatoid arthritis patients treated with infliximab. AB - INTRODUCTION: The objective of our study was to identify the significance of the subtypes of dendritic cell (DC), specifically myeloid DCs (mDCs) and plasmacytoid DCs (pDCs), in rheumatoid arthritis (RA) pathogenesis through their longitudinal follow-up in patients receiving infliximab. METHODS: Circulating mDC and pDC levels were evaluated by flow cytometry in RA patients (n = 61) and healthy volunteers (n = 30). In RA patients, these levels were measured before and during infliximab therapy. Their counts were correlated to RA disease activity markers and anti-nuclear antibody occurrence. IFNalpha production was measured by ELISA in serum of RA patients and, in vitro, in supernatant of peripheral blood mononuclear cells stimulated by influenza virus in the presence or absence of infliximab. Statistical evaluations were based on Mann-Whitney tests or Wilcoxon's signed-rank tests. RESULTS: RA patients with active disease were characterized by a baseline decrease in both circulating pDCs and mDCs. Disease activity markers inversely correlated only with mDC level. This level increased in RA patients responsive to infliximab therapy, to reach the level observed in controls. Conversely, anti-nuclear antibody appearance during infliximab therapy correlated inversely with pDC level and was associated with increased serum IFNalpha level and circulating plasma cells number. In vitro studies revealed that infliximab kept pDCs in an IFNalpha secreting state upon viral stimulation allowing differentiation of B cells into anti-nuclear antibody-secreting plasma cells. CONCLUSIONS: This study reveals two distinct roles for pDC and mDC in RA. Circulating mDCs mainly contribute to RA activity, whereas pDCs seem to be involved in appearance of anti-nuclear antibodies under infliximab therapy through the ability of this drug to keep pDCs in an IFNalpha secreting state. PMID- 19563674 TI - Optimum land cover products for use in a Glossina-morsitans habitat model of Kenya. AB - BACKGROUND: Tsetse flies are the primary vector for African trypanosomiasis, a disease that affects both humans and livestock across the continent of Africa. In 1973 tsetse flies were estimated to inhabit 22% of Kenya; by 1996 that number had risen to roughly 34%. Efforts to control the disease were hampered by a lack of information and costs associated with the identification of infested areas. Given changing spatial and demographic factors, a model that can predict suitable tsetse fly habitat based on land cover and climate change is critical to efforts aimed at controlling the disease. In this paper we present a generalizable method, using a modified Mapcurves goodness of fit test, to evaluate the existing publicly available land cover products to determine which products perform the best at identifying suitable tsetse fly land cover. RESULTS: For single date applications, Africover was determined to be the best land use land cover (LULC) product for tsetse modeling. However, for changing habitats, whether climatically or anthropogenically forced, the IGBP DISCover and MODIS type 1 products where determined to be most practical. CONCLUSION: The method can be used to differentiate between various LULC products and be applied to any such research when there is a known relationship between a species and land cover. PMID- 19563673 TI - Management of the critically poisoned patient. AB - BACKGROUND: Clinicians are often challenged to manage critically ill poison patients. The clinical effects encountered in poisoned patients are dependent on numerous variables, such as the dose, the length of exposure time, and the pre existing health of the patient. The goal of this article is to introduce the basic concepts for evaluation of poisoned patients and review the appropriate management of such patients based on the currently available literature. METHODS: An unsystematic review of the medical literature was performed and articles pertaining to human poisoning were obtained. The literature selected was based on the preference and clinical expertise of authors. DISCUSSION: If a poisoning is recognized early and appropriate testing and supportive care is initiated rapidly, the majority of patient outcomes will be good. Judicious use of antidotes should be practiced and clinicians should clearly understand the indications and contraindications of antidotes prior to administration. PMID- 19563675 TI - Enhanced detection method for corneal protein identification using shotgun proteomics. AB - BACKGROUND: The cornea is a specialized transparent connective tissue responsible for the majority of light refraction and image focus for the retina. There are three main layers of the cornea: the epithelium that is exposed and acts as a protective barrier for the eye, the center stroma consisting of parallel collagen fibrils that refract light, and the endothelium that is responsible for hydration of the cornea from the aqueous humor. Normal cornea is an immunologically privileged tissue devoid of blood vessels, but injury can produce a loss of these conditions causing invasion of other processes that degrade the homeostatic properties resulting in a decrease in the amount of light refracted onto the retina. Determining a measure and drift of phenotypic cornea state from normal to an injured or diseased state requires knowledge of the existing protein signature within the tissue. In the study of corneal proteins, proteomics procedures have typically involved the pulverization of the entire cornea prior to analysis. Separation of the epithelium and endothelium from the core stroma and performing separate shotgun proteomics using liquid chromatography/mass spectrometry results in identification of many more proteins than previously employed methods using complete pulverized cornea. RESULTS: Rabbit corneas were purchased, the epithelium and endothelium regions were removed, proteins processed and separately analyzed using liquid chromatography/mass spectrometry. Proteins identified from separate layers were compared against results from complete corneal samples. Protein digests were separated using a six hour liquid chromatographic gradient and ion-trap mass spectrometry used for detection of eluted peptide fractions. The SEQUEST database search results were filtered to allow only proteins with match probabilities of equal or better than 10-3 and peptides with a probability of 10-2 or less with at least two unique peptides isolated within the run along with default Xcorr values. These parameters resulted in the identification of over 350 proteins, including over 225 new proteins not previously detected in the cornea by mass spectrometry. In addition, corneal layer separation resulted in identification of nearly every protein that was identified in the complete cornea assay. The epithelium and endothelium each revealed many unique proteomes specific to each layer. In the endothelium, the protein olfactomedin-like 3 was identified for the first time in the cornea by this analysis. Olfactomedin-3 is a neuronal expressed protein also known as optimedin that stimulates formation of cell adherent and cell-cell tight junctions and its expression modulates cytoskeleton organization and cell migration. However, the function of this protein in rabbit corneal endothelium is currently unknown. CONCLUSION: This manuscript presents a description of a more comprehensive proteomic profile for mammalian cornea compared to past methods. The use of simple dissection procedures of the tissue and the application of long chromatographic gradients, many more proteins can be identified. PMID- 19563676 TI - Comparison of metal-dependent catalysis by HIV-1 and ASV integrase proteins using a new and rapid, moderate throughput assay for joining activity in solution. AB - BACKGROUND: HIV-1 integrase (IN) is an attractive target for the development of drugs to treat AIDS, and inhibitors of this viral enzyme are already in the clinic. Nevertheless, there is a continuing need to devise new approaches to block the activity of this viral protein because of the emergence of resistant strains. To facilitate the biochemical analysis of wild-type IN and its derivatives, and to measure the potency of prospective inhibitory compounds, a rapid, moderate throughput solution assay was developed for IN-catalyzed joining of viral and target DNAs, based on the detection of a fluorescent tag. RESULTS: A detailed, step-by-step description of the new joining assay is provided. The reactions are run in solution, the products captured on streptavidin beads, and activity is measured by release of a fluorescent tag. The procedure can be scaled up for the analysis of numerous samples, and is substantially more rapid and sensitive than the standard radioactive gel methods. The new assay is validated and its utility demonstrated via a detailed comparison of the Mg++- and Mn++ dependent activities of the IN proteins from human immunodeficiency virus type 1 (HIV-1) and the avian sarcoma virus (ASV). The results confirm that ASV IN is considerably more active than HIV-1 IN, but with both enzymes the initial rates of joining, and the product yields, are higher in the presence of Mn++ than Mg++. Although the pH optima for these two enzymes are similar with Mn++, they differ significantly in the presence of Mg++, which is likely due to differences in the molecular environment of the binding region of this physiologically relevant divalent cation. This interpretation is strengthened by the observation that a compound that can inhibit HIV-1 IN in the presence of either metal cofactors is only effective against ASV in the presence of Mn++. CONCLUSION: A simplified, assay for measuring the joining activity of retroviral IN in solution is described, which offers several advantages over previous methods and the standard radioactive gel analyses. Based on comparisons of signal to background ratios, the assay is 10-30 times more sensitive than gel analysis, allows more rapid and accurate biochemical analyses of IN catalytic activity, and moderate throughput screening of inhibitory compounds. The assay is validated, and its utility demonstrated in a comparison of the metal-dependent activities of HIV-1 and ASV IN proteins. PMID- 19563677 TI - In silico genomic analyses reveal three distinct lineages of Escherichia coli O157:H7, one of which is associated with hyper-virulence. AB - BACKGROUND: Many approaches have been used to study the evolution, population structure and genetic diversity of Escherichia coli O157:H7; however, observations made with different genotyping systems are not easily relatable to each other. Three genetic lineages of E. coli O157:H7 designated I, II and I/II have been identified using octamer-based genome scanning and microarray comparative genomic hybridization (mCGH). Each lineage contains significant phenotypic differences, with lineage I strains being the most commonly associated with human infections. Similarly, a clade of hyper-virulent O157:H7 strains implicated in the 2006 spinach and lettuce outbreaks has been defined using single-nucleotide polymorphism (SNP) typing. In this study an in silico comparison of six different genotyping approaches was performed on 19 E. coli genome sequences from 17 O157:H7 strains and single O145:NM and K12 MG1655 strains to provide an overall picture of diversity of the E. coli O157:H7 population, and to compare genotyping methods for O157:H7 strains. RESULTS: In silico determination of lineage, Shiga-toxin bacteriophage integration site, comparative genomic fingerprint, mCGH profile, novel region distribution profile, SNP type and multi-locus variable number tandem repeat analysis type was performed and a supernetwork based on the combination of these methods was produced. This supernetwork showed three distinct clusters of strains that were O157:H7 lineage-specific, with the SNP-based hyper-virulent clade 8 synonymous with O157:H7 lineage I/II. Lineage I/II/clade 8 strains clustered closest on the supernetwork to E. coli K12 and E. coli O55:H7, O145:NM and sorbitol-fermenting O157 strains. CONCLUSION: The results of this study highlight the similarities in relationships derived from multi-locus genome sampling methods and suggest a "common genotyping language" may be devised for population genetics and epidemiological studies. Future genotyping methods should provide data that can be stored centrally and accessed locally in an easily transferable, informative and extensible format based on comparative genomic analyses. PMID- 19563678 TI - Predicting protein-protein interactions in Arabidopsis thaliana through integration of orthology, gene ontology and co-expression. AB - BACKGROUND: Large-scale identification of the interrelationships between different components of the cell, such as the interactions between proteins, has recently gained great interest. However, unraveling large-scale protein-protein interaction maps is laborious and expensive. Moreover, assessing the reliability of the interactions can be cumbersome. RESULTS: In this study, we have developed a computational method that exploits the existing knowledge on protein-protein interactions in diverse species through orthologous relations on the one hand, and functional association data on the other hand to predict and filter protein protein interactions in Arabidopsis thaliana. A highly reliable set of protein protein interactions is predicted through this integrative approach making use of existing protein-protein interaction data from yeast, human, C. elegans and D. melanogaster. Localization, biological process, and co-expression data are used as powerful indicators for protein-protein interactions. The functional repertoire of the identified interactome reveals interactions between proteins functioning in well-conserved as well as plant-specific biological processes. We observe that although common mechanisms (e.g. actin polymerization) and components (e.g. ARPs, actin-related proteins) exist between different lineages, they are active in specific processes such as growth, cancer metastasis and trichome development in yeast, human and Arabidopsis, respectively. CONCLUSION: We conclude that the integration of orthology with functional association data is adequate to predict protein-protein interactions. Through this approach, a high number of novel protein-protein interactions with diverse biological roles is discovered. Overall, we have predicted a reliable set of protein-protein interactions suitable for further computational as well as experimental analyses. PMID- 19563679 TI - Intrinsic bias in breast cancer gene expression data sets. AB - BACKGROUND: While global breast cancer gene expression data sets have considerable commonality in terms of their data content, the populations that they represent and the data collection methods utilized can be quite disparate. We sought to assess the extent and consequence of these systematic differences with respect to identifying clinically significant prognostic groups. METHODS: We ascertained how effectively unsupervised clustering employing randomly generated sets of genes could segregate tumors into prognostic groups using four well characterized breast cancer data sets. RESULTS: Using a common set of 5,000 randomly generated lists (70 genes/list), the percentages of clusters with significant differences in metastasis latencies (HR p-value<0.01) was 62%, 15%, 21% and 0% in the NKI2 (Netherlands Cancer Institute), Wang, TRANSBIG and KJX64/KJ125 data sets, respectively. Among ER positive tumors, the percentages were 38%, 11%, 4% and 0%, respectively. Few random lists were predictive among ER negative tumors in any data set. Clustering was associated with ER status and, after globally adjusting for the effects of ER-alpha gene expression, the percentages were 25%, 33%, 1% and 0%, respectively. The impact of adjusting for ER status depended on the extent of confounding between ER-alpha gene expression and markers of proliferation. CONCLUSION: It is highly probable to identify a statistically significant association between a given gene list and prognosis in the NKI2 dataset due to its large sample size and the interrelationship between ER-alpha expression and markers of proliferation. In most respects, the TRANSBIG data set generated similar outcomes as the NKI2 data set, although its smaller sample size led to fewer statistically significant results. PMID- 19563680 TI - Rationale, design and methods for a randomised and controlled trial to investigate whether home access to electronic games decreases children's physical activity. AB - BACKGROUND: Many children are reported to have insufficient physical activity (PA) placing them at greater risk of poor health outcomes. Participating in sedentary activities such as playing electronic games is widely believed to contribute to less PA. However there is no experimental evidence that playing electronic games reduces PA. There is also no evidence regarding the effect of different types of electronic games (traditional sedentary electronic games versus new active input electronic games) on PA. Further, there is a poor understanding about how characteristics of children may moderate the impact of electronic game access on PA and about what leisure activities are displaced when children play electronic games. Given that many children play electronic games, a better understanding of the effect of electronic game use on PA is critical to inform child health policy and intervention. METHODS: This randomised and controlled trial will examine whether PA is decreased by access to electronic games and whether any effect is dependent on the type of game input or the child's characteristics. Children aged 10-12 years (N = 72, 36 females) will be recruited and randomised to a balanced ordering of 'no electronic games', 'traditional' electronic games and 'active' electronic games. Each child will participate in each condition for 8 weeks, and be assessed prior to participation and at the end of each condition. The primary outcome is PA, assessed by Actical accelerometers worn for 7 days on the wrist and hip. Energy expenditure will be assessed by the doubly labelled water technique and motor coordination, adiposity, self-confidence, attitudes to technology and PA and leisure activities will also be assessed. A sample of 72 will provide a power of > 0.9 for detecting a 15 mins difference in PA (sd = 30 mins). DISCUSSION: This is the first such trial and will provide critical information to understand whether access to electronic games affects children's PA. Given the vital importance of adequate PA to a healthy start to life and establishing patterns which may track into adulthood, this project can inform interventions which could have a profound impact on the long term health of children. TRIAL REGISTRATION: This trial is registered in the Australia and New Zealand Clinical Trials Registry (ACTRN 12609000279224). PMID- 19563682 TI - DNA fingerprinting differentiation between beta-carotene hyperproducer strains of Dunaliella from around the world. AB - BACKGROUND: Dunaliella salina is the most important species of the genus for beta carotene production. Several investigations have demonstrated that D. salina produces more than 10% dry weight of pigment and that the species grows in salt saturated lagoons. High plasticity in the green stage and the almost indistinguishable differences in the red phase make identification and differentiation of species and ecotypes very difficult and time consuming. RESULTS: In this work, we applied our intron-sizing method to compare the 18S rDNA fingerprint between D. salina (CCAP 19/18), D. salina/bardawil (UTEX LB2538) and beta-carotene hyperproducing strains of Dunaliella isolated from salt saturated lagoons in Baja, Mexico. All hyperproducer strains reached beta carotene levels of about 10 pg/cell. Optical microscopy did not allow to differentiate between these Dunaliella strains; however, 18S rDNA fingerprinting methodology allowed us to differentiate D. salina from D. salina/bardawil. CONCLUSION: In Baja Mexico we found D. salina and D. salina/bardawil species by using intron-sizing-method. The National Center for Biotechnology Information (NCBI) Dunaliella 18S rDNA gene sequences were analyzed with our methodology and extraordinary correlation was found with experimental results. PMID- 19563681 TI - Age-related associations of hypertension and diabetes mellitus with chronic kidney disease. AB - BACKGROUND: Studies suggest end-stage renal disease incidence and all-cause mortality rates among patients with chronic kidney disease (CKD) differ by age. The association of diabetes mellitus and hypertension with CKD across the adult lifespan is not well established. METHODS: Data from NHANES 1999-2004 were used to determine the association of risk factors for stage 3 or 4 CKD (n = 12,518) and albuminuria (n = 12,778) by age grouping (20 to 49, 50 to 69, and > or =70 years). Stage 3 or 4 CKD was defined as an estimated glomerular filtration rate of 15 to 59 ml/min/1.73 m2 and albuminuria as an albumin to creatinine ratio > or =30 mg/g. RESULTS: For adults 20 to 49, 50 to 69 and > or =70 years of age, the prevalence ratios (95% confidence interval) of stage 3 or 4 CKD associated with hypertension were 1.94 (0.86 - 4.35), 1.51 (1.09 - 2.07), 1.31 (1.15 - 1.49), respectively (p-trend = 0.038). The analogous prevalence ratios (95% confidence interval) were 3.01 (1.35 - 6.74), 1.61 (1.15 - 2.25), 1.40 (1.15 - 1.69), respectively, for diagnosed diabetes mellitus (p-trend = 0.067); and 2.67 (0.53 - 13.4), 1.35 (0.69 - 2.63), 1.08 (0.78 - 1.51), respectively, for undiagnosed diabetes mellitus (p-trend = 0.369). The prevalence ratios of albuminuria associated with hypertension and diagnosed and undiagnosed diabetes mellitus were lower at older age (each p < 0.05). CONCLUSION: Among US adults, diabetes mellitus and hypertension are associated with CKD and albuminuria regardless of age. However, the associations were stronger at younger ages. PMID- 19563683 TI - Expression of therapeutic misconception amongst Egyptians: a qualitative pilot study. AB - BACKGROUND: Studies have shown that research participants fail to appreciate the difference between research and medical care, labeling such phenomenon as a "therapeutic misconception" (TM). Since research activity involving human participants is increasing in the Middle East, qualitative research investigating aspects of TM is warranted. Our objective was to assess for the existence of therapeutic misconception amongst Egyptians. METHODS: Study Tool: We developed a semi-structured interview guide to elicit the knowledge, attitudes, and perspectives of Egyptians regarding medical research. SETTING: We recruited individuals from the outpatient settings (public and private) at Ain Shams University in Cairo, Egypt. ANALYSIS: Interviews were taped, transcribed, and translated. We analyzed the content of the transcribed text to identify the presence of a TM, defined in one of two ways: TM1 = inaccurate beliefs about how individualized care can be compromised by the procedures in the research and TM2 = inaccurate appraisal of benefit obtained from the research study. RESULTS: Our findings showed that a majority of participants (11/15) expressed inaccurate beliefs regarding the degree with which individualized care will be maintained in the research setting (TM1) and a smaller number of participants (5/15) manifested an unreasonable belief in the likelihood of benefits to be obtained from a research study (TM2). A total of 12 of the 15 participants were judged to have expressed a TM on either one of these bases. CONCLUSION: The presence of TM is not uncommon amongst Egyptian individuals. We recommend further qualitative studies investigating aspects of TM involving a larger sample size distinguished by different types of illnesses and socio-economic variables, as well as those who have and have not participated in clinical research. PMID- 19563684 TI - Role of accelerated segment switch in exons to alter targeting (ASSET) in the molecular evolution of snake venom proteins. AB - BACKGROUND: Snake venom toxins evolve more rapidly than other proteins through accelerated changes in the protein coding regions. Previously we have shown that accelerated segment switch in exons to alter targeting (ASSET) might play an important role in its functional evolution of viperid three-finger toxins. In this phenomenon, short sequences in exons are radically changed to unrelated sequences and hence affect the folding and functional properties of the toxins. RESULTS: Here we analyzed other snake venom protein families to elucidate the role of ASSET in their functional evolution. ASSET appears to be involved in the functional evolution of three-finger toxins to a greater extent than in several other venom protein families. ASSET leads to replacement of some of the critical amino acid residues that affect the biological function in three-finger toxins as well as change the conformation of the loop that is involved in binding to specific target sites. CONCLUSION: ASSET could lead to novel functions in snake venom proteins. Among snake venom serine proteases, ASSET contributes to changes in three surface segments. One of these segments near the substrate binding region is known to affect substrate specificity, and its exchange may have significant implications for differences in isoform catalytic activity on specific target protein substrates. ASSET therefore plays an important role in functional diversification of snake venom proteins, in addition to accelerated point mutations in the protein coding regions. Accelerated point mutations lead to fine-tuning of target specificity, whereas ASSET leads to large-scale replacement of multiple functionally important residues, resulting in change or gain of functions. PMID- 19563685 TI - The effects of gender and age on health related behaviors. AB - BACKGROUND: Lifestyle-related diseases, including diabetes, cardiovascular disease, and some cancers represent the greatest global health threat. Greater insight into health needs and beliefs, using broad community samples, is vital to reduce the burden of chronic disease. This study aimed to investigate gender, age, screening practices, health beliefs, and perceived future health needs for healthy ageing. METHODS: Random probability sampling using self-completion surveys in 1456 adults residing in Australia. RESULTS: Screening behaviors were associated with gender and age. Men and women >51 years were more likely (27%) to have screening health checks than those <50 years (2%). Factors nominated to influence health were lifestyle (92%), relationships (82%), and environment (80%). Women were more likely to nominate preparedness to have an annual health check, willingness to seek advice from their medical practitioner and to attend education sessions. Numerous health fears were associated with ageing, however participants were more likely to have a financial (72%) rather than a health plan (42%). More women and participants >51 years wanted information regarding illness prevention than men or those aged <30 years. CONCLUSION: Age and gender are associated with health related behaviors. Optimal health is perceived as a priority, yet often this perception is not translated into preventative action. These findings will inform future research and policy makers as we strive towards a healthier ageing society and the prevention of chronic disease. PMID- 19563688 TI - Incessant pericardial effusion in a 9 year old male responding to infliximab. AB - Recurrent idiopathic pericardial effusion can be a challenging medical problem. Multiple medical interventions may yield minimal improvement. We describe a patient with an incessant pericardial effusion that responded to infliximab. The use of infliximab should be considered in the management of recurrent pericardial effusions resistant to other treatment modalities. PMID- 19563686 TI - PKCepsilon-dependent potentiation of TTX-resistant Nav1.8 current by neurokinin-1 receptor activation in rat dorsal root ganglion neurons. AB - BACKGROUND: Substance P (SP), which mainly exists in a subtype of small-diameter dorsal root ganglion (DRG) neurons, is an important signal molecule in pain processing in the spinal cord. Our previous results have proved the expression of SP receptor neurokinin-1 (NK-1) on DRG neurons and its interaction with transient receptor potential vanilloid 1 (TRPV1) receptor. RESULTS: In this study we investigated the effect of NK-1 receptor agonist on Na(v)1.8, a tetrodotoxin (TTX)-resistant sodium channel, in rat small-diameter DRG neurons employing whole cell patch clamp recordings. NK-1 agonist [Sar(9), Met(O2)(11)]-substance P (Sar SP) significantly enhanced the Na(v)1.8 currents in a subgroup of small-diameter DRG neurons under both the normal and inflammatory situation, and the enhancement was blocked by NK-1 antagonist Win51708 and protein kinase C (PKC) inhibitor bisindolylmaleimide (BIM), but not the protein kinase A (PKA) inhibitor H89. In particular, the inhibitor of PKCepsilon, a PKC isoform, completely blocked this effect. Under current clamp model, Sar-SP reduced the amount of current required to evoke action potentials and increased the firing rate in a subgroup of DRG neurons. CONCLUSION: These data suggest that activation of NK-1 receptor potentiates Na(v)1.8 sodium current via PKCepsilon-dependent signaling pathway, probably participating in the generation of inflammatory hyperalgesia. PMID- 19563689 TI - Short-term and long-term outcomes for congenital cardiac surgery. PMID- 19563687 TI - Isolation of a human-like antibody fragment (scFv) that neutralizes ricin biological activity. AB - BACKGROUND: Ricin is a lethal toxin that inhibits protein synthesis. It is easily extracted from a ubiquitously grown plant, Ricinus communis, and thus readily available for use as a bioweapon (BW). Anti-ricin antibodies provide the only known therapeutic against ricin intoxication. RESULTS: In this study, after immunizing a non-human primate (Macaca fascicularis) with the ricin chain A (RTA), a phage-displayed immune library was built (2 x 108 clones), that included the lambda light chain fragment. The library was screened against ricin, and specific binders were sequenced and further analyzed. The best clone, 43RCA, was isolated using a new, stringent neutralization test. 43RCA had a high, picomolar affinity (41 pM) and neutralized ricin efficiently (IC50 = 23 +/- 3 ng/ml, corresponding to a [scFv]/[ricin] molar ratio of 4). The neutralization capacity of 43RCA compared favourably with that of polyclonal anti-deglycosylated A chain (anti-dgRCA) IgGs, obtained from hyperimmune mouse serum, which were more efficient than any monoclonal at our disposal. The 43RCA sequence is very similar to that for human IgG germline genes, with 162 of 180 identical amino acids for the VH and VL (90% sequence identity). CONCLUSION: Results of the characterization studies, and the high degree of identity with human germline genes, altogether make this anti-ricin scFv, or an IgG derived from it, a likely candidate for use in humans to minimize effects caused by ricin intoxication. PMID- 19563690 TI - Treatment of monozygotic twins with obsessive compulsive disorder using cognitive therapy and exposure with ritual prevention. AB - BACKGROUND: Assessment and treatment of cohabiting monozygotic (MZ) twins with obsessive compulsive disorder (OCD) is a possible challenge for clinical psychologists. AIMS: This study aims to present a detailed history of two sets of cohabiting MZ twins with OCD, and describe the adaption of cognitive behavior therapy in their treatments. METHOD: Two sets of cohabiting MZ twins completed a structured intake and the Yale Brown Obsessive Compulsive Scale as well as measures of depression, anxiety, a measure of obsessive beliefs, and thought action fusion. One set received cognitive behavior therapy and exposure with ritual prevention (ERP) simultaneously and the other received ERP separately. Pre , post-treatment and follow-up assessments occurred for both sets of twins. RESULTS: All four individuals showed notable decreases in OCD, and results were maintained for three of the four participants at follow-up points. CONCLUSIONS: This study highlights the developmental course of OCD that can occur in cohabiting twins, and the clinical adaption that may be necessary. PMID- 19563691 TI - The impact of age on the postprandial vascular response to a fish oil-enriched meal. AB - Although chronic fish oil intervention had been shown to have a positive impact on vascular reactivity, very little is known about their acute effects during the postprandial phase. Our aim was to examine the impact of a fish oil-enriched test meal on postprandial vascular reactivity in healthy younger ( < 50 years) v. older ( > or = 50 years) men. Vascular reactivity was measured at baseline (0 h), 2 and 4 h after the meal by laser Doppler iontophoresis and blood samples taken at 0 and 4 h for the measurement of plasma lipids, total nitrite, glucose and insulin. Acetylcholine- (ACh, endothelial-dependent vasodilator) and sodium nitroprusside (SNP, endothelial-independent vasodilator)-induced reactivities were greater at 4 h than at baseline or 2 h in the younger men (P < 0.04). These changes were not observed in the older men. Comparison of the male groups revealed significantly greater responses to ACh (P = 0.006) and SNP (P = 0.05) at 4 h in the younger compared with the older males. Postprandial NEFA concentrations were also greater at 4 h in the younger compared with the older men (P = 0.005), with no differences observed for any of the other analytes. Multiple regression analysis revealed age to be the most significant predictor of both ACh and SNP induced reactivity 4 h after the meal. In conclusion, the ingestion of a meal enriched in fish oil fatty acids was shown to improve postprandial vascular reactivity at 4 h in our younger men, with little benefit evident in our older men. PMID- 19563692 TI - Effects of pomace olive oil-enriched diets on endothelial function of small mesenteric arteries from spontaneously hypertensive rats. AB - Pomace olive oil (POM), an olive oil subproduct traditionally used in Spain, is a good source of minor components from the unsaponifiable fraction such as triterpenoids, mainly in the form of oleanolic acid, which induces vascular protection and vasodilatation. Our aim was to evaluate the effects of long-term intake of diets enriched in POM with high concentration in oleanolic acid on endothelial dysfunction associated to hypertension in small mesenteric arteries (SMA) from spontaneously hypertensive rats (SHR). During 12 weeks, rats (six rats per group) were fed either a control 2 % maize oil diet (BD), or high-fat diets containing 15 % refined olive oil (OL), pomace olive oil (POM), or pomace olive oil supplemented in oleanolic acid (POMO; up to 800 parts per million). Endothelial and vascular functions were assessed by relaxing or contracting responses to acetylcholine (ACh) or phenylephrine, respectively. The involvement of endothelium-derived relaxing factors in these responses was evaluated. In contrast to BD, SHR fed high-fat diets showed a biphasic response to ACh related to changes in eicosanoid metabolism. POM enhanced the endothelial function in SMA from SHR by increasing the endothelium-derived hyperpolarising factor (EDHF)-type component, whereas administration of POMO resulted in a similar contribution of NO/EDHF in the endothelial response to ACh. The present study shows that despite the lack of changes in blood pressure, consumption of POM improves endothelial function in SMA from SHR by improving the agonist-mediated EDHF/NO response. Thus, triterpenoids confer a protective role to POM against endothelial dysfunction in hypertension. PMID- 19563694 TI - Gender difference in health and its determinants in the old-aged population in India. AB - This paper examines the gender differential in health and its socioeconomic and demographic determinants in the old-age population of India based on the National Sample Survey 60th round data collected in 2004. As in developed countries, older women in India report poorer self-reported health and experience greater immobility compared with men. Stepwise logistic regression analysis shows that the gender differential in health is linked to various socioeconomic and demographic variables and that the gender gap could be narrowed with appropriate policy intervention. Specifically, paying special attention towards improving the socioeconomic status of widowed/separated women could attenuate a substantial portion of the observed gender gap in the health of the old-age population. PMID- 19563693 TI - Lactobacillus helveticus and Bifidobacterium longum taken in combination reduce the apoptosis propensity in the limbic system after myocardial infarction in a rat model. AB - Myocardial infarction (MI) stimulates the release of pro-inflammatory substances that induce apoptosis in the limbic system. Pro-inflammatory cytokines are considered as the root cause of apoptosis, although the mechanism is not fully explained and/or understood at this time. In addition, depression may induce gastrointestinal perturbations that maintain the elevated levels of pro inflammatory cytokines. It has been shown that some specific probiotic formulations may reduce gastrointestinal problems induced by stress and the pro/anti-inflammatory cytokine ratio. Therefore, we hypothesised that probiotics, when given prophylactically, may diminish the apoptosis propensity in the limbic system following a MI. Male adult Sprague-Dawley rats were given probiotics (Lactobacillus helveticus and Bifidobacterium longum in combination) or placebo in their drinking-water for four consecutive weeks. A MI was then induced in the rats by occluding the left anterior coronary artery for 40 min. Rats were killed following a 72 h reperfusion period. Infarct size was not different in the two groups. Bax/Bcl-2 (pro-apoptotic/anti-apoptotic) ratio and caspase-3 (pro apoptotic) activity were reduced in the amygdala (lateral and medial), as well as in the dentate gyrus in the probiotics group when compared with the placebo. Akt activity (anti-apoptotic) was increased in these same three regions. No significant difference was observed in Ca1 and Ca3 for the different markers measured. In conclusion, the probiotics L. helveticus and B. longum, given in combination as preventive therapy, reduced the predisposition of apoptosis found in different cerebral regions following a MI. PMID- 19563695 TI - Maternal smoking and acute respiratory infection symptoms among young children in Nepal: multilevel analysis. AB - The association between maternal smoking and adverse child health outcomes has not been systematically explored in less developed countries, especially in Nepal where over a quarter of women of reproductive age smoke tobacco products. This study aims to quantify the effect of maternal smoking on acute respiratory infection (ARI) symptoms among children aged below five years, using the 2001 Nepal Demographic and Health Survey. It is hypothesized that children born to mothers who smoke frequently are at higher risk of developing ARI symptoms. Four level random intercept logistic regression models were used to disentangle the independent effect of maternal smoking on children's ARI symptoms, controlling for potential biological, socioeconomic, seasonal and spatial variables. Maternal smoking status had a significant effect on children's ARI symptoms; the effects were significantly higher (adjusted OR 1.41; 95% CI 1.02-1.96) among those born to mothers who smoked more frequently than their counterparts. Furthermore, a strong spatial pattern was evident in the prevalence of ARI symptoms, after adjusting for maternal smoking and relevant control variables. The findings underscore the importance of designing exclusive public health intervention measures to prevent tobacco smoking within households, for example through awareness campaigns highlighting the adverse effect of maternal smoking on child health. PMID- 19563696 TI - Prevalence of hepatitis C virus infection in patients with COPD. AB - Some studies have suggested that chronic hepatitis C virus (HCV) infection may induce an accelerated decline of forced expiratory volume in 1 second (FEV(1)). We performed a cross-sectional study to determine the prevalence of HCV infection in a sample of chronic obstructive pulmonary disease (COPD) patients and in a control group of blood donors. The clinical characteristics of HCV-positive and HCV-negative patients were compared. Anti-HCV antibody was determined and confirmed by HCV-RNA. The prevalence of HCV infection in COPD patients was 7.5% (95% CI 6.52-8.48) and in blood donors was 0.41% (95% CI 0.40-0.42). The HCV positive patients had a lower FEV(1) (34.7 +/- 8.6%) and a higher BODE index (median = 6) than HCV-negative patients (42.7 +/- 16.5%, median = 4, respectively) (P = 0.011 and 0.027, respectively). Our results suggest a high prevalence of chronic HCV infection in patients with COPD in comparison with the blood donors. HCV-positive patients have a more severe disease. PMID- 19563697 TI - Environmental and ecological potential for enzootic cycles of Puumala hantavirus in Great Britain. AB - Puumala virus (PUUV) is a zoonotic rodent-borne hantavirus in continental Europe. Its reservoir host, the bank vole (Myodes glareolus), is ubiquitous in Great Britain (GB); however, there has been no reported incidence of virus in either animals or humans. In northwest Europe, increases in bank vole numbers, stimulated by increases in production of beech/oak crops (mast), are associated with outbreaks of nephropathia epidemica (NE) in humans. These so-called 'mast years' are determined by sequential climatic events. This paper investigates the contribution of a number of ecological and environmental factors driving outbreaks of PUUV in northwest Europe and assesses whether such factors might also permit enzootic PUUV circulation in GB. Analysis of GB climate data, using regression models, confirms that mast years in GB are stimulated, and can be predicted, by the same climatic events as mast years in PUUV-endemic regions of northwest Europe. A number of other possible non-climatic constraints on enzootic cycles are discussed. PMID- 19563698 TI - National mortality rates from chronic liver disease and consumption of alcohol and pig meat. AB - A correlation between national pig-meat consumption and mortality rates from chronic liver disease (CLD) across developed countries was reported in 1985. One possible mechanism explaining this may be hepatitis E infection spread via pig meat. We aimed to re-examine the original association in more recent international data. Regression models were used to estimate associations between national pig-meat consumption and CLD mortality, adjusting for confounders. Data on CLD mortality, alcohol consumption, hepatitis B virus (HBV) and hepatitis C virus (HCV) seroprevalence for 18 developed countries (1990-2000) were obtained from WHO databases. Data on national pig-meat and beef consumption were obtained from the UN database. Univariate regression showed that alcohol and pig-meat consumption were associated with mortality from CLD, but beef consumption, HBV and HCV seroprevalence were not. A 1 litre per capita increase in alcohol consumption was associated with an increase in mortality from CLD in excess of 1.6 deaths/100,000 population. A 10 kg higher national annual average per capita consumption of pork meat was associated with an increase in mortality from CLD of between 4 and 5 deaths/100,000 population. Multivariate regression showed that alcohol, pig-meat consumption and HBV seroprevalence were independently associated with mortality from CLD, but HCV seroprevalence was not. Pig-meat consumption remained independently associated with mortality from CLD in developed countries in the 1990-2000 period. Further work is needed to establish the mechanism. PMID- 19563699 TI - Quinolone-resistant Salmonella Typhi in South Africa, 2003-2007. AB - In South Africa, for the years 2003-2007, the Enteric Diseases Reference Unit received 510 human isolates of Salmonella Typhi, of which 27 were nalidixic acid resistant [minimum inhibitory concentrations (MICs) 128-512 microg/ml] with reduced susceptibility to ciprofloxacin (MICs 0.125-0.5 microg/ml). Pulsed-field gel electrophoresis analysis of 19 available isolates differentiated them into five DNA pattern types; multiple-locus variable-number tandem repeat analysis differentiated the isolates into 10 types. This level of genetic diversity suggested that resistant strains usually emerged independently of one another. A 16- to 32-fold decrease in nalidixic acid MIC and a 2- to 8-fold decrease in ciprofloxacin MIC, was observed in the presence of an efflux pump inhibitor. All isolates were negative by PCR screening for qnr genes. Seven resistant isolates were further analysed for mutations in the quinolone resistance-determining region of gyrA, gyrB, parC and parE. No amino-acid mutations were identified in GyrB and ParE; all isolates showed amino-acid mutations in both GyrA and ParC. We conclude that amino-acid mutations in GyrA and ParC in combination with active efflux of antibiotic out of the bacterial cell are the probable mechanisms conferring quinolone resistance. PMID- 19563701 TI - [Attaching importance to clinical study of neo-adjuvant chemoradiotherapy followed by surgery for esophageal squamous cell carcinoma]. PMID- 19563702 TI - [Isolation and identification of cancer stem cells from human esophageal carcinoma]. AB - OBJECTIVE: To isolate and identify cancer stem cells from esophageal carcinoma cells (ECCs) using cell surface marker p75NTR. METHODS: ECCs cell lines were established with ECCs collected from 38 surgically resected specimens. Flow cytometry was used to identify the p75NTR positive cells therein that were isolated then using magnetic activated cell sorting (MACS) method. The growing characteristics in DMEM medium and capability of colony-forming in soft agar of the p75NTR positive cells were evaluated ex vivo with MTT method. p75NTR positive cells of different concentrations were subcutaneously injected into the backs of Balb/c nude mice and PBS was injected into the contralateral back, and then tumorigenesis was observed, 8 weeks later the mice were killed with their tumors taken out to undergo microscopy. RESULTS: Eight ECCs cell lines were established, 6 of which were found to contain 0.32%-3.35% of p75NTR positive cells. The purity of p75NTR positive cells isolated by MACS was up to 90%. MTT result showed that population doubling time of the p75NTR positive cells was (17+/-3) hours, significantly shorter than that of the p75NTR negative cells [(37+/-7) hours, P<0.01]. The colony-forming rate in soft agar of the p75NTR positive cells was (45.9%+/-8.9%), significantly higher than that of the p75NTR negative cells [(3.7%+/-2.1%), P<0.01]. As few as 2000 p75NTR positive cells gave rise to new tumors in xenotransplantation, with a tumorigenic ability 50 times as high as that of the p75NTR negative cells. CONCLUSION: p75NTR positive cells carry some properties of cancer stem cells, such as the ability of self-renewal, differentiation and proliferation and demonstrate higher ability of colony forming ex vivo and tumorigenesis in vivo. PMID- 19563703 TI - [Prevention and treatment of complications after surgical resection for esophageal and gastric cardiac cancers]. AB - OBJECTIVE: To summarize the experience in surgical resection for esophageal and gastric cardiac cancers so as to put forward countermeasures to prevent the post operative complications. METHODS: From September 1952 to December 2005, 20,796 patients with esophageal and gastric cardiac cancers underwent surgical operation. The category and incidence of the complications in different decades were retrospectively analyzed. RESULTS: (1) 18,772 of the 20 796 patients with esophageal and gastric cardiac cancers underwent surgical resection. Operative complications occurred in 1741 patients (9.27%), death occurred in 433 of which (24.87%). (2) The complication rate was 39.77% in 1950s, and decreased to 4.10% in 2000s. The mortality rate of complication was 44.29% in 1950s, and decreased to 15.42% in 2000s. (3) The incidence and mortality rates of closely-surgery related complications declined obviously over the period of study. The incidence and mortality rates of anastomotic leakage were 4.55% and 50.00% respectively in 1950s, and then decreased to 1.21% and 3.33% respectively in 2000s. The incidence and the mortality rates of empyema were 7.39% and 38.46% respectively in 1950s, and decreased to 0.36% and 5.56% respectively in 2000s. The incidence and the mortality rates of chylothorax was 0.16% and 0% recently. (4) The incidence of non-surgical-related complications declined slightly but the mortality rate was still very high over the period of study. The mortality rates of pulmonary and cardiovascular complication were 27.42% and 25.00% respectively recently. CONCLUSION: (1) One of the advantages of surgical treatment for esophagus and gastric cardiac cancers is the obvious decline of the incidence and the mortality rates of complications. (2) In order to reduce the incidence of systemic complication, especially pulmonary complication, active and corresponding preoperative preparation should be improved along with the expansion of the scope of surgical indication. (3) The key for prevention of post-operative complications is consummate surgery techniques. Prevention and treatment of anastomotic leakage, gastric wall necrosis, and intrathoracic and intraabdominal bleeding are still the focal points. PMID- 19563704 TI - [Surgical treatment of esophageal carcinoma with cervical esophagogastrostomy by left thoracic esophagectomy]. AB - OBJECTIVE: To evaluate the effects of surgical treatment with cervical esophagogastrostomy by left thoracic esophagectomy on esophageal carcinoma. METHODS: The clinical data of 3169 patients with esophageal carcinoma, 2115 males and 1054 females, aged 60.9 (30-82), that underwent cervical esophagogastrostomy by left thoracic esophagectomy from Jan. 1990 to Dec. 2000 were analyzed retrospectively. RESULTS: The incidence of complication was 8.6% (273). The operative mortality was 0.6% (19). The 1, 3, and 5 year survival rates were 81.5%, 51.6%, and 32.1% respectively. Manometric examination of the digestive tract showed that the post-operative resting pressure of residual esophagus above the anastomotic stoma was (16+/-11 mm Hg), significantly higher than that of the cervical esophagus of the normal controls [(4+/-3) mm Hg), P<0.01]. CONCLUSION: Reducing incidence of gastroesophageal reflux and improving the quality of life of patients, cervical anastomosis should be recommended as the first choice procedure in treatment of upper and middle segment esophageal carcinoma. PMID- 19563700 TI - Molecular mechanisms of endothelial hyperpermeability: implications in inflammation. AB - Endothelial hyperpermeability is a significant problem in vascular inflammation associated with trauma, ischaemia-reperfusion injury, sepsis, adult respiratory distress syndrome, diabetes, thrombosis and cancer. An important mechanism underlying this process is increased paracellular leakage of plasma fluid and protein. Inflammatory stimuli such as histamine, thrombin, vascular endothelial growth factor and activated neutrophils can cause dissociation of cell-cell junctions between endothelial cells as well as cytoskeleton contraction, leading to a widened intercellular space that facilitates transendothelial flux. Such structural changes initiate with agonist-receptor binding, followed by activation of intracellular signalling molecules including calcium, protein kinase C, tyrosine kinases, myosin light chain kinase, and small Rho-GTPases; these kinases and GTPases then phosphorylate or alter the conformation of different subcellular components that control cell-cell adhesion, resulting in paracellular hypermeability. Targeting key signalling molecules that mediate endothelial junction-cytoskeleton dissociation demonstrates a therapeutic potential to improve vascular barrier function during inflammatory injury. PMID- 19563705 TI - [Characteristics of gene structure in facioscapulohumeral muscular dystrophy related 4q35 subtelomere and genotype-phenotype correlation in Chinese Han population]. AB - OBJECTIVE: To investigate the characteristics of gene structure in facioscapulohumeral muscular dystrophy (FSHD)-related 4q35 subtelomere, to analyze the distribution of 2 alleles (4qA and 4qB) distal to D4Z4 of this locus, and to further elucidate the genotype-phenotype correlation in Chinese Han FSHD patients. METHODS: Peripheral blood samples were collected from 52 unrelated families including 62 FSHD-affected and 57 unaffected members. Genomic DNA was extracted from the lymphocytes according to the specific procedure designed to minimize DNA shearing, then digested with EcoRI or HindIII, or double digested with EcoRI and BlnI. The cleaved DNA was separated by pulsed field electrophoresis (PFGE) and Southern blotting with the probes p13E-11, 4qA, and 4qB. The size of FSHD-causing 4qA allele and its distribution was analyzed by "curve fitting". Then the characteristics of translocation and mosaicism, the frequencies of two allelic variants of chromosome 4q and their genotypes were calculated to analyze the genotype-phenotype correlation. RESULTS: It was found that 69 patients carried a short chromosome 4-type allele of 4qA origin with the length 10-38 kb. The mean length of these pathogenic EcoRI/4qA arrays was 20 kb+/ 7 kb, without significant difference between the sporadic cases and familial cases (t=1.413, P>0.05). Three different translocation types were observed with a translocation rate of 14.49%. The rate of 4q-->10q translocation was 13.04%, significantly higher than that of 10q-->4q translocation (1.45%, chi2=6.900, P<0.05). Somatic mosaicism was detected in a male sporadic case and a female asymptomatic familial case. In 57 cases with standard configuration distribution, the frequency of 4qA/4qB heterozygote was 61.40%, significantly higher than that of 4qA/4qA homozygote (38.60%, (chi2=5.930, P<0.05). There were not significant differences in the repeat size distributions and assessment of clinical severity between the sporadic and familial cases (t=-0.039, P>0.05; H=0.693, P>0.05). CONCLUSION: About 95% of Chinese FSHD patients carry a pathogenic 4-type allele of 4qA origin less than 30 kb long. The frequent translocations between chromosome 4q and 10q may play an essential role for FSHD mechanism. The frequency of 4qA/4qB heterozygote is significantly higher than that of 4qA/4qA homozygote, while the allelic variant genotypes do not contribute to modify FSHD manifestations. PMID- 19563706 TI - [Three-dimensional numerical simulation and hemodynamic analysis of intracranial longitypical aneurysms]. AB - OBJECTIVE: To establish a model of three-dimensional numerical simulation of intracranial longitypical aneurism and analyze the hemodynamic features thereof. METHODS: Ten patients with intracranial longitypical aneurism underwent surgical treatment. Intracranial vascular ultrasonography was conducted before operation. Intra-operatively three-dimensional angiography of the aneurysm was conducted and multifunctional physiological monitor was used to record the dynamic pressure and pressure wave form of the main branch parent artery. Matlab, Ansys, and Fluent software were used to simulate the blood flow of the longitypical aneurysms. RESULTS: The hemodynamic parameter levels were the highest at the inflow tract, followed by those at the outflow tract, and were the lowest at the top of aneurysm, for example, the blood flow velocity was (1.07+/-0.23) m/s, the dynamic pressure was (574+/-186) Pa, and the wall shear stress was (7.7+/-2.0) Pa at the inflow tract, all significantly higher than those at the top [(0.15+/-0.07) m/s, (37+/-13) Pa, and (0.40+/-0.13) Pa respectively, all P<0.05]. No eddy or just simple eddy occurred in the aneurysm. CONCLUSION: The dynamic pressure, velocity, and wall shear stress are the lowest at the top of longitypical aneurysm which may contribute to the rupture of aneurysm. PMID- 19563708 TI - [Clinical analysis of intraocular pressure elevation following vitreoretinal surgery]. AB - OBJECTIVE: To study the incidence and risk factors of intraocular pressure (IOP) elevation in patients who underwent vitreoretinal surgery. METHODS: Data for 422 patients (446 eyes) who underwent vitreoretinal surgery from June 2005 to June 2007 were retrospectively analyzed. IOP was measured before surgery and on day 1, 2, 3, 7-14 after surgery with Goldmann applanation tonometer. Ocular hypertension was defined as IOP 25 mmHg or more. All factors were analyzed for association with Ocular hypertension. RESULTS: 232 of the 422 patients were female. 190 were male. The IOP was elevated significantly in 185 eyes (41%) after surgery within 2 weeks. Among them the IOP occurred mostly in day 1 (152 eyes, 82%). The rate of IOP elevation with intraocular tamponade of C3F8 or silicone oil was 53% or 55%, the difference being not statistically significant (P>0.05). The incidence of high IOP in C3F8 injection group and silicone oil injection group were statistically higher than simple pars plana vitrectomy group (P<0.01). The risk factors of ocular hypertension include C3F8 injection (chi2=37.82, P<0.01), silicone oil injection (chi2=27.84, P<0.01), lentectomy (chi2=8.98, P<0.01), scatter endolaser (chi2=7.41, P<0.01), diabetes (chi2=12.12, P<0.01) and increasing patient age (chi2=7.07, P<0.01). CONCLUSION: IOP elevation was a common complication of vitreoretinal surgery. The risk factors of ocular hypertension include C3F8 injection, silicone oil injection, lentectomy, scatter endolaser, diabetes and increasing patient age. PMID- 19563707 TI - [Effects of patient-controlled analgesia with small dose ketamine combined with morphine and the influence thereof on plasma beta-endorphin level in patients after radical operation for esophageal carcinoma]. AB - OBJECTIVE: To evaluate the effects of patient-controlled analgesia (PCA) with small dose ketamine combined with morphine on analgesia and influence thereof on the plasma beta-endorphin (EP) level in the patients after radical operation for esophageal carcinoma. METHODS: Thirty ASAI-II patients, aged 35-65, weighing 42 75 kg, with visual analogue score>or=3, undergoing elective radical operation for esophageal carcinoma under general anesthesia received intravenous morphine 2 - 3 mg were randomly divided into 2 equal groups: group m receiving morphine 0.02 mg.kg(-1).h(-1), and with group mk receiving morphine 0.02 mg.kg(-1).h(-1) combined with ketamine 0.08 mg.kg(-1).h(-1) for 50 h. In the course of treatment the patients received intravenous injection of morphine 2-3 mg when the VAS was >or=3. VAS was recorded 4, 8, 20, 24, and 48 h after operation. The amount of morphine used after operation, PCA button pressing times (effective/active), side effects, and vital signs including pulse, saturation of blood oxygen, respiratory rate, heart rate, and average arterial pressure were recorded. Central venous blood samples were collected when entering the operation room (T0), by the end of operation (T1), and 6 h (T2), 24 h (T3), and 48 h (T4) after operation respectively to examine the beta-endorphin level. RESULTS: During the period 4-48 h after operation the VAS scores of the group mk were significantly lower than those of the group m in activity state (all P<0.05) and were not significantly different those of the group m at resting state (all P>0.05). The total amount of morphine consumed and the actual PCA button pressing times were significantly less in the group mk than in the group m (both P<0.05). The incidence rates of nausea, vomiting, and pruritus of the group mk were all significantly lower than those of the group m (all P<0.05). There were not significant differences in the incidence rates of dreaming and pseudoesthesia between these 2 groups. All the vital signs were stable in the 2 groups. The plasma beta-EP levels at the time point T1 of these 2 groups were both significantly higher than those at T0 (both P<0.05). The plasma beta-endorphin levels at T2-4 of the group mk decreased gradually from the level at T1 to the level at T0, and the plasma beta-endorphin levels of the group m rapidly decreased from the level at T0 to the T0 level and remained at this level to the 48 h after operation. CONCLUSION: The combination of small dose of ketamine with morphine provides optimal analgesia with low side effect rate and little effect on the plasma beta-EP level. PMID- 19563709 TI - [Short- to mid-term evaluation of CT-guided 125I brachytherapy on recurrent or metastatic head and neck cancers]. AB - OBJECTIVE: to evaluate the short- and medium-term clinical effects of 125I seed implantation on recurrent or metastatic head and neck cancers. METHODS: Thirty patients with recurrent or metastatic head and neck cancers after operation, radiotherapy, or chemotherapy, totaling 421 lesions 4.2 (2-9) cm in diameter, 23 males and 12 females, aged 56 (39-71), underwent implantation of 12-70 125I seeds (on average 33 per person) under the guidance of CT, ultrasonography, or endoscopy with an interval of 1 cm between any 2 seeds with the radioactive activity per seed of 29.6 MBq and matched peripheral dose of 90-160 Gy. Follow-up was conducted for 13 (4-40) months to observe the local control rate, overall survival rate, pain relief, and clinical complications. RESULTS: Follow-up 4 months later showed that 24 of the 42 lesions obtained complete remission, 11 obtained partial remission, 5 no change, and 2 progress of disease, with a clinical response rate of 83.3% (CR+PR). The overall 1-, 2-, and 3-year survival rates were 88.4%, 72.4%, and 45.2% respectively with a median survival time of 31 months. The pain relief rate was 73.4% (17/23). The long-term complications included hyperpigmentation at operative sites (n=4), insensible feeling on lateral cheek (n=3), dryness of oral cavity (n=2), and headache combined with infection (n=1). CONCLUSION: Relieving the pain, improving the life quality, CT guided radioactive 125I seed implantation is a simple, safe, and effective method in treating recurrent or metastatic head and neck cancer with minimal damage and few complications. PMID- 19563710 TI - [Epidemiology of bacterial dysentery diagnosed clinically in Beijing: analysis of 652 patients]. AB - OBJECTIVE: To investigate the epidemiological and clinical characteristics of acute bacterial dysentery in recent years in Beijing so as to provide scientific reference for prevention and control of enteric infectious diseases. METHODS: The data of age, occupation, symptoms, signs, pathogen detection of 652 patients with acute bacterial dysentery diagnosed clinically from April 2007 to October 2007 were analyzed retrospectively. RESULTS: The occupations of the 652 patients included office workers and functionaries (31.4%), students (29.4%), retirees (9.8%), unemployed and housewives (7.1%), commercial attendants (6.3), workers (4.8%), teachers (4.1%), and others (7.1%). Diarrhea, stomach ache, and fever were the main symptoms. Tenesmus and tenderness of left lower quadrant abdomen were infrequent. The positive rate of fecal bacterial culture was 17.7%. 8.9% of the patients were diagnosed as with bacterial dysentery by fecal bacterial culture. CONCLUSION: Most of the patients with bacterial dysentery in Beijing are young adults. The positive rate of fecal bacterial culture is low. It is difficult to differentiate bacterial dysentery from other diarrheal diseases. PMID- 19563711 TI - [A clinical, electromyographic, and magnetic resonance imaging study of cervical spondylotic myelopathy: analysis of 96 cases]. AB - OBJECTIVE: To investigate the clinical, electromyographic, and magnetic resonance imaging (MRI) manifestations of the patients with cervical spondylotic myelopathy (CSM) and the correlation among them. METHOD: 96 CSM patients, 56 males and 40 females, aged 53 (2/32-72), underwent electromyography (EMG) of the deltoid muscle (C5), biceps muscle of arm (C6), common extensor muscle of fingers (C7), and short abductor muscle of thumb and abductor muscle of little finger (C8-T), and MRI of the cervical vertebrae before surgical treatment. The clinical symptoms were graded according to the Japanese Orthopaedic Association (JOA) scoring system. The correlation among the JOA score, MIR classification, and EMG manifestation was analyzed. RESULTS: EMG showed that 61 of the 96 patients (64%) with root injury at C5, 6, 7, and 8. Those positive in EMG showed higher severity in MRI (Z=2.863, P=0.004). The JOA score was not correlated with the degree of spinal cord compression demonstrated by MRI and the EMG results. CONCLUSION: The root injury rate is high in the CSM patients. The patients positive in EMG have severe MRI results. No clear correlation is between the severity of clinical symptom and MRI of cervical vertebrae and root injury demonstrated by EMG. Hence, treatment should be considered by integration of EMG, MRI and JOA scores. PMID- 19563712 TI - [Intracranial aneurysms difficult to be treated by endovascular interventional therapy: report of 27 patients]. AB - OBJECTIVE: To analyze the characteristics of aneurysms difficult to be treated by endovascular interventional therapy and report the results of operation. METHODS: 27 patients with 33 intracranial aneurysms transferred from the ward of interventional therapy, 7 males and 20 females, aged 47 (27-67), accepted microsurgical operation, The neck, volume, aspect ratio and blood vessel condition of the aneurysms were analyzed. RESULTS: The aneurysms difficult to be treated by endovascular interventional therapy included those with wide neck (>4 mm), large volume (maximal diameter>or=8 mm), and failure in endovascular maneuvers (because of flexed blood vessel and severe vasospasm). The total outcome of operation was good and no death occurred. CONCLUSION: Some aneurysms difficult to be treated by endovascular interventional therapy should be treated by microsurgical operation, which will get a better result. PMID- 19563713 TI - [Treatment of ureteral obstruction by holmium: YAG laser endoureterotomy: a report of 18 cases]. AB - OBJECTIVE: To investigate the clinical value and safety of holmium: YAG laser endoureterotomy in the treatment of ureteral obstruction. METHODS: Holmium: YAG laser endoureterotomy, with the laser optic fiber 550 microm in diameter and the output power of 3.5 Watt, via ureteroscopy, was performed on 18 patients ureteral obstruction, 8 males and 10 females, aged 52.1 (34-67), 11 with the stricture in the upper segment (complete obstruction in 4 cases), 5 in the middle segment, and 2 in lower segment; and 6 cases complicated with ureteral calculus. Postoperatively, an orthopedic ureteral stent (a 6-Fr double-J ureteral stent with a movable 5 cm length 9-Fr orthopedic cannula) was remained indwelling for 3 6 months. Follow-up was conducted for 10.7 (2-14) months. RESULTS: The operative duration was 32 (25-70) minutes. One patient underwent failed endoureterotomy and was turned to percutaneous nephroscopy. Success was achieved in 16 patients. The glomerular filtration rate (GFR) of these affected kidneys increased from 16.4+/ 6.9 ml/min ante-operatively to 24.9+/-8.2 ml/min (P<0.01) postoperatively. One kidney was resected because of non-function, with GFR of 2 ml/min and intractable pyelitis. No recurrence of ureteral stricture was observed. CONCLUSION: Holmium: YAG laser endoureterotomy with insertion of orthopedic ureteral stent is an efficient and safe treatment for ureteral strictures with minimal invasion, less complications and easy recovery. This operation should be performed with a thorough preparation and severely restricted indication. PMID- 19563714 TI - [Treatment of early osteonecrosis of femoral head by core decompression and autograft of mesenchymal stem cell cultured in vitro under arthroscopy: experiment with rabbits]. AB - OBJECTIVE: To observe the therapeutic effect of core decompression and autograft of mesenchymal stem cells (MSCs) cultured in vitro under arthroscopy in treatment of early osteonecrosis of femoral head. METHODS: Thirty-six rabbits were made into models of early osteonecrosis of femoral head of the right leg, and then randomly divided into 3 equal groups: Group A group as control group, Group B group undergoing simple core decompression under arthroscopy by clearing out of necrotic bone, and Group C undergoing core decompression and implantation of MSCs. Four and 8 weeks later 6 rabbits from each group underwent roentgenography and then were killed with their femoral heads taken out to undergo histological examination. RESULTS: Four weeks later new bone was evident in the implantation field of Group C while fibrous tissues were evident in the femoral head of Group B. Pathological examination showed regular shape and normal density of femoral head in Group C and irregular shape and low density of femoral head in Group B. X ray photograph 8 weeks after treatment showed that the femoral head articular surface of Group A collapsed, uneven density and defect of spongy bone were seen in Group B, and the shape of femoral head of Group C was regular without collapse and the original defect had been repaired. CONCLUSION: With strong osteogenic function, MSCs improve the repair of ischemic necrosis of femoral head and can be used in treatment of early osteonecrosis of femoral head via core decompression and autograft of MSCs cultured in vitro under arthroscopy. PMID- 19563715 TI - [Low dose X-irradiation promotes callus mineralization: experiment with rats]. AB - OBJECTIVE: To investigate the effect of low dose irradiation (LDI) on callus formation and mineralization. METHODS: 80 SD rats were subjected to standard closed fracture on right femur so as to establish animal models and then were randomly divided into 2 equal groups: LDI group undergoing low dose whole body X ray irradiation of 1Gy right after fracture induction and control group without irradiation. 1, 2, 3, 4, and 8 weeks later blood samples were collected from the heart to examine the amounts of white blood cells and platelets and alkaline phosphatase (ALP) level. Conventional X-ray films were taken and MicroCT was conducted to evaluate the callus bridging. Then the rats were killed to take out the femurs to undergo histological examination. Peripheral quantitative computed tomography (pQCT) was conducted to quantify the bone mineral content (BMC). Four point bending test was used to examine the mechanical properties of the callus. RESULTS: The maximum load value and histological score 2 weeks later of the LDI group were both lower than those of the control group, however, the serum ALP, BMC, maximum load level, callus bridging score, and histological score 3 weeks later of the LDI group were all significantly higher than those of the control groups (all P<0.05). 4 and 8 weeks later there were not significant differences in all these parameters between the 2 groups. The WBC amount in the first 2 weeks of experiment of the LDI group was remarkably lower in these 2 groups, however, in the third week, the WBC value of the LDI group was higher than that of the control group (P<0.05). CONCLUSION: LDI promotes the mineralization at the stage of hard callus formation which is probably associated with progenitor cell mobilization. PMID- 19563716 TI - [Transfection of siRNA expressing plasmids targeting S gene inhibits replication and expression of hepatitis B virus in hepatic cancer cells]. AB - OBJECTIVE: To study the inhibitive effects of transfection of siRNA expressing plasmids targeting S gene, one of the 4 open reading frames of hepatitis B virus (HBV), on the replication and expression of HBV. METHODS: Two plasmids expressing 2 siRNAs targeting S gene, one of the 4 open reading frames of HBV (S1 and S2) and one nonspecific plasmid (siRNA-S3), as negative control, with the length of 21 nt heterologous to the HBV/U95551 genome were constructed, and then transfected into the hepatic cancer cells of the line HepG2.2.15. 48 hours later, real-time PCR was used to evaluate the gene silencing efficiency and ELISA was used to detect the expression of HBsAg and hepatitis B e antigen (HBeAg), protein markers of HBV, in the supernatants. RESULTS: The inhibition rates of HBsAg and HBeAg expression of the HepG2.2.15 cells transfected with S1 were 60% and 56% respectively, those of the HepG2.2.15 cells transfected with S2 were 73% and 70% respectively, those of the HepG2.2.15 cells transfected with S1+S2 were 82% and 78% respectively, and those of the HepG2.2.15 cells transfected with S3 were not significantly different from those of the blank control group. RT-PCR showed that the mRNA expression rates of HBsAg and HBeAg in the HepG2.2.15 cells transfected with S1, S2, and S1+S2 were inhibited by 64%-88% t respectively. CONCLUSION: Transfection of the vector plasmids expressing the siRNAs targeting S gene inhibits the expression of HBsAg and HBeAg in hepatic cancer cells. RNAi may provide a viable strategy against viruses for the prevention and treatment of HBV infection. PMID- 19563717 TI - [Effect of omega-3 long-chain polyunsaturated fatty acids on the cultured oligodendrocyte precursor cells]. AB - OBJECTIVE: To explore the effect of long-chain polyunsaturated fatty acids (LPFA) on the survival and process growth of the brain hippocampal Oligodendrocyte precursor cells (OPC). METHOD: Two kinds of cultured neural progenitor cells isolated from adult rat hippocampus were used. After arachidonic acid (AA), eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) treatment respectively, the activity of cells were determined by Lactate dehydrogenase (LDH) assay, and the quantitative measurements of the cell processes were done after the fluorescent immune cells staining. RESULTS: EPA, DHA and AA showed similar trends to increase the cell numbers when the concentrations were high (50 micromol/L, P<0.05), but only EPA and DHA elongated the process of OPC significantly (P<0.05). CONCLUSION: omega-3 LPFA (EPA and DHA) could enhance the survival and process growth of OPC. PMID- 19563718 TI - [The determination of medical reference values for tumor markers in cerebrospinal fluid]. AB - OBJECTIVE: To determine medical reference values for tumor markers in cerebrospinal fluid. METHODS: Concentrations of CEA, CA125, CA15-3, CA19-9, CA72 4, CYFRA21-1, AFP, NSE, SCC and HCG were determined by means of double-antibody sandwich ELISA in 110 patients excluding primary tumors and meningeal carcinomatosis using Roche E170 modular immunoassay analyzer. RESULTS: The determined medical reference values for tumor biomarkers in cerebrospinal fluid were as follows: CEA<0.573 microg/L, CA125<2.591 U/ml, CA15-3<2.045 U/ml, CA19 9<2.272 U/ml, CA72-4<1.252 U/ml, CYFRA21-1<1.44 ng/ml, AFP<0.968 microg/L, NSE<57.666 ng/ml, SCC<0.5 microg/L, HCG<0.769 U/L. There was no correlation between any tumor marker and age (P>0.05). Concentrations of tumor markers were not affected by gender (P>0.05). CONCLUSION: Medical reference values for CEA, CA125, CA15-3, CA19-9, CA72-4, CYFRA21-1, AFP, NSE, SCC and HCG in cerebrospinal fluid were first determined. PMID- 19563721 TI - Refining the role of antiplatelet therapy for atrial fibrillation. PMID- 19563722 TI - Cardioverter-defibrillators for primary and secondary prevention in adults with tetralogy of fallot. PMID- 19563723 TI - Advances in percutaneous coronary intervention. AB - Initially developed more than 30 years ago as simple balloon-based dilatation for focal coronary stenoses, the introduction of the coronary stent allowed percutaneous coronary intervention (PCI) to be applied to most patients with coronary artery disease. PCI improves quality of life by relieving angina in patients with stable coronary artery disease and can be life saving in patients with extensive ischemia and acute coronary syndromes. The safety of the PCI procedure has been enhanced significantly by the appropriate application of adjunctive pharmacotherapy, most importantly antithrombotic and antiplatelet agents. The advent of drug-eluting stents has significantly prolonged the durability of PCI. The safety and efficacy of PCI continue to evolve with the development of next-generation stents and safer and improved pharmacologic agents. An extensive evidence base of randomized clinical trials has established PCI as an important strategy with a pivotal role in the management of patients with coronary artery disease. PMID- 19563724 TI - New noninvasive imaging technologies in coronary artery disease. AB - Coronary artery disease affects a large population. Recent emphasis on primary and secondary prevention has made an impact on the detection of atherosclerosis, yet the incidence of acute coronary syndromes continues to increase. This has steered the cardiology community toward improving and developing new imaging techniques that are capable of detecting disease at a very early preclinical state. Coronary CT angiography is capable of characterizing plaques and detecting eccentric lesions that would not appear on stress testing or cardiac catheterization. Cardiac MRI provides high-resolution imaging of plaques in addition to tissue characterization without the ionizing radiation associated with other imaging techniques. Positron emission tomography is a rapidly growing imaging tool that detects inflammation associated with coronary atherosclerosis. In the near future, these new noninvasive modalities will play an intricate part in primary prevention and in diagnosis and treatment follow-up. PMID- 19563725 TI - Cardiovascular disease and type 2 diabetes mellitus: regulating glucose and regulating drugs. AB - Type 2 diabetes mellitus is a major and increasingly prevalent independent risk factor for cardiovascular morbidity and mortality worldwide. Glycemic control is a target of therapy and a principal marker of therapeutic success in diabetes, but whether lowering glucose is accompanied by a commensurate reduction in cardiovascular risk is a matter of ongoing controversy. It has become increasingly apparent from recent large-scale clinical outcome trials that glucose lowering is a poor predictor of cardiovascular outcome, and several instances of unexpectedly increased cardiovascular risk with antihyperglycemic drugs have sounded the alarm with regulatory agencies. This article reviews the critical facts that have led to a recent shift in the regulation of glucose lowering drugs and makes the case for why new and existing antidiabetic medications should be assessed in clinical trials of cardiovascular outcome. PMID- 19563726 TI - Medical therapy in acute coronary syndromes: which medicines and at what doses? AB - Acute coronary syndrome (ACS) occurs when plaque rupture in a coronary artery is superimposed with thrombus formation. This accounts for 1.7 million hospital admissions in the United States annually and significant morbidity and mortality. Although there are advantages to an invasive approach to treating patients with ACS, the role of medical therapy is vital as an adjunctive treatment to reperfusion therapies and in stabilizing ruptured plaques and modifying the metabolic milieu that predisposes to plaque formation and rupture. This article reviews the most important drug classes for medical treatment of ACS patients, as well as optimal doses. This is an exciting time to be involved in the field of cardiovascular medicine, as we continue to see profound improvement from medical therapy in the morbidity and mortality associated with ACS. PMID- 19563727 TI - Lipid reduction in acute coronary syndrome: how much, when, and how? AB - The use of statins for secondary prevention after acute coronary syndrome is well established. In recent years, trials have investigated the dose of statins used and timing of administration. Initiation of statin therapy as early as 1 day after an acute coronary syndrome event has been shown to be effective in reducing major adverse cardiovascular events. The benefit of early statin use is linked to reduction in inflammation and increased compliance with therapy. In addition, intensive therapy further reduces events and inflammation, as reflected by decreased C-reactive protein. Given the findings of these recent studies, early and intensive lipid-lowering therapy with a statin is justified and safe. PMID- 19563728 TI - Genomic view of factors leading to plaque instability. AB - The manifestations of coronary artery disease are varied. They all arise as a consequence of the deposition of atherosclerotic plaque within the vessel wall. The most feared sequela of coronary artery disease is sudden and unexpected death in the ostensibly healthy patient. Plaque rupture of hemodynamically insignificant atherosclerotic plaques and ensuing thrombosis is likely responsible for a large proportion of such deaths. Identifying populations at increased risk for sudden death would represent a major advance. Such screening is contingent upon identification of DNA sequence variants that predispose individuals to plaque rupture. Phenotyping is not sufficiently nuanced to detect such variants on a large scale, so we are limited to end points that are crude surrogates for plaque rupture. As imaging modalities are refined and our ability to recruit large numbers of appropriate patients is facilitated by the formation of alliances, our ability to probe this conundrum via a genome-wide approach will improve. PMID- 19563729 TI - Transition and transfer from pediatric to adult care of young adults with complex congenital heart disease. AB - Many children with complex congenital heart disease are now surviving childhood with the potential to live meaningful and productive adult lives. The process of transitioning or preparing patients and families for transfer from pediatric to adult care is challenging and rarely implemented properly. An inadequate transition process results in delayed and inappropriate care, improper timing of transfer, and undue emotional and financial stress on the patients, their families, and the health care system. At worst, patients are lost to appropriate follow-up. This article discusses the general principles of transition and transfer for young adults with chronic illness, highlights the needs of young adults with congenital heart disease, discusses the barriers to transition, and proposes goals and key elements of a formal transition program. PMID- 19563730 TI - Choosing the best contraceptive method for the adult with congenital heart disease. AB - As women with congenital heart disease (CHD) approach childbearing age, issues such as pregnancy and contraception need to be addressed. Women with CHD represent a heterogeneous population. While some have simple cardiac lesions and thus have contraceptive risks that are no different from those of the general population, others have complex, multisystem disease such that incorrect contraception advice can be associated with serious health consequences. Choosing a contraceptive requires consideration of the risk of pregnancy; the available contraception options; their risks, failure rates, and benefits; and the preferences of the woman. This review focuses on contraceptive selection in women with CHD. PMID- 19563731 TI - Percutaneous interventions in adults with congenital heart disease: expanding indications and opportunities. AB - Percutaneous intervention in adults with congenital heart disease has advanced rapidly in recent years and represents a growing field of invasive cardiology. Valve replacement, stent implantation for native and recurrent coarctation, and ventricular septal defect closure have emerged as attractive alternatives to surgery and have been added to the existing validated procedures (eg, atrial septum defect closure, persistent ductus arteriosus occlusion, and pulmonary valve dilatation). This review summarizes current and expanding indications for transcatheter interventions in adults with congenital heart disease. PMID- 19563733 TI - Effect of Zanthoxylum schinifolium on TNF-alpha-induced vascular inflammation in human umbilical vein endothelial cells. AB - Pro-inflammatory cytokines induce the injury of endothelial cells in response to increases of adhesion molecules, leading to vascular inflammation and the development of atherosclerosis. In this study, we evaluated an ethanol extract of Zanthoxylum schinifolium (EZS) to determine if it inhibits the expressions of cellular adhesion molecules in human umbilical vein endothelial cells (HUVEC). When pretreatment of HUVEC with EZS, EZS suppressed the expression levels of cell adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-), and E-selectin induced by TNF-alpha. The adhesion of HL-60 cells to TNF-alpha-induced endothelial cells was decreased significantly in a concentration-dependent manner. Furthermore, TNF-alpha-induced MCP-1 and IL-8 mRNA expression levels were also attenuated by pretreatment with EZS. In addition, EZS suppressed TNF-alpha-induced production of reactive oxygen species (ROS). EZS inhibited NF-kappaB activation and IkappaB-alpha phosphorylation induced by TNF-alpha, subsequent degradation of IkappaB-alpha. Finally, EZS inhibited TNF-alpha-induced p38 MAPK and c-Jun N-terminal kinase (JNK) phosphorylation. Taken together, these results demonstrate that EZS suppresses vascular inflammatory process, which may be closely related to the inhibition of ROS, JNK, p38 MAPK and NF-kappaB activation in HUVEC. PMID- 19563732 TI - Pulmonary hypertension complicating congenital heart disease. AB - Congenital heart disease can predispose individuals to pulmonary vascular remodeling as a result of the abnormality in pulmonary blood flow and pressure that accompanies the specific congenital defect being considered. Pulmonary arterial hypertension associated with congenital heart defects is an important determinant of functional capacity and survival, especially when the Eisenmenger's state of reversed shunt is present. The likelihood of right ventricular dysfunction and failure increases with the degree of pulmonary artery pressure. Thus, the aim of disease management in this patient population should be to prevent or improve right heart failure. Current therapies that modify the progression of pulmonary vascular disease-including endothelin-1 receptor antagonists, phosphodiesterase-5 inhibitors, and prostanoids-should be considered carefully in patients with congenital heart disease-associated pulmonary hypertension. The risks and benefits of altering the balance of pulmonary vascular resistance to systemic vascular resistance must be weighed for each patient. PMID- 19563734 TI - IL-18 accelerates the cell apoptosis by up-regulating cysteinyl leukotriene 2 receptor expression in human umbilical vein endothelial cells at the early stage of administration. AB - The purpose of the present study is to identify whether interleukin (IL)-18 can modulate cysteinyl leukotriene 2 receptor (CysLT2R) expression in Human Umbilical Vein Endothelial Cells (HUVECs) and how it influences the cell death. According to the results from real-time reverse transcription PCR, confocal laser scanning microscopy, and western blotting, a dose-dependent augmentation of CysLT2R protein expression in HUVECs was triggered by IL-18 for the first 2 h followed by down-regulation within the next 22 h after IL-18 administration. The flow cytometry showed that non-selective CysLT1R and CysLT2R antagonist BAY-u9773 could attenuate the early stage apoptosis mediated by IL-18 whereas CysLT1R antagonist Montelukast couldn't. Also, pretreatment with BAY-u9773 suppressed calcium influx of HUVECs induced by IL-18 whereas Montelukast didn't work. The observation that progression of cell death aggravated by IL-18 could be attenuated by BAY-u9773 may offer a chance to develop a novel way to treat arteriosclerosis. PMID- 19563735 TI - A combination of Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, improved renal microvascular insufficiency in 5/6 nephrectomized rats. AB - Chronic renal ischemia and hypoxia in the tubulointerstitium are involved in the mechanisms of progressive chronic kidney disease. Previous studies showed that the decoction of a combination of two Chinese herbs, Astragalus membmnaceus var. mongholicus and Angelica sinensis (A & A) has antifibrotic effects through multiple pathways in different animal models. In this study, remnant kidney model was employed to investigate whether A & A affect the expression of VEGF, the density of the renal microvasculature and thus alleviate the renal injury. Rats were divided randomly into four groups: sham group (N-31), 5/6 Nx group (5/ 6 nephrectomy, N=43), A & A treated group (A & A group, N=40, A & A 12 g/kg/d po), enalapril treated group (Ena group, N=56, enalapril 4 mg/kg/d po). Rats from each group were sacrificed at the 2th, 4th, 8th and 12th weeks respectively after surgery and treatment The 24 h urinary protein excretion, serum creatinine (Scr) and urea were measured. The collagen IV (COL-IV), fibronectin (FN), aminopeptidase P (APP) and VEGF were stained using immunohistochemistry. The COL IV, FN and APP were semi-quantitatively analyzed. Peritubular capillary density in the cortical interstitial area was quantified. The level of VEGF was assayed by ELISA. The results revealed that Scr, urea and urinary protein excretion remained constant at each time point in sham group. Compared to sham group, 5/6 Nx group was shown severe glomerulosclerosis, tubulointerstitial lesions and vascular damage, as well as higher level of Scr from the 2nd week (72.3 +/- 5.2 vs. 48.6 +/- 2.6 micromol/L P < 0.05) to the 12th week (71.9 +/- 8.0 vs. 55.7 +/- 4.5 micromol/L P < 0.05). Although there was no significant difference in Scr level after treatment of enalapril or A & A (P > 0.05), kidney damage was alleviated at the 8th and the 12th week in the two treatment groups (P < 0.05, vs. 5/6 Nx group). The urinary protein excretion of 5/6 Nx group was significantly increased from the 4th week, it was 1.5, 2.4 and 3.8 fold of that of sham group at the 4th, 8th and 12th week, respectively. Compared to 5/6 Nx group, proteinuria was decreased by enalapril to 59%, 33% at 8th and 12th week. After A & A administration, urinary protein excretion decreased to 66%, 56%, 75%, 55% of 5/6 Nx group at the 2nd, 4th, 8th and 12th, respectively (P < 0.05). Compared with sham group, there was increased expression of FN and COL-IV in rats with 5/6 Nx. After A & A or enalapril administration, the expression of FN and COL-IV was significantly decreased compared with that in the 5/6 Nx group at 8th and 12th week (P < 0.05). On the other hand, the capillary density was decreased at the 8th and 12th week in 5/6 Nx rats (P < 0.01). In A & A-treated group, similarly with enalapril group, the amount of APP-positive glomerular capillary increased by 36% (P < 0.01), and the peritubular capillary density was increased 94% at 8th week and 52% at 12th week compared with 5/6 Nx group (P < 0.05). The renal level of VEGF was decreased in 5/6 Nx rats, but increased at the 8th and 12th week in A & A group (P < 0.05, vs. 5/6 Nx group). In conclusion, A & A has renoprotective effects by suppression of extra cellular matrix deposition in 5/6 Nx rat. The renoprotective effects may be associated with reduction of proteinuria, up-regulation of VEGF which may reduce the loss of capillaries and improve microstructure dysfunction. PMID- 19563736 TI - Signaling pathways involved in DNA synthesis and migration in response to lysophosphatidic acid and low-density lipoprotein in coronary artery smooth muscle cells. AB - Low-density lipoprotein (LDL) and lysophosphatidic acid (LPA), one of the lipid components of lipoprotein, induced the DNA synthesis of coronary artery smooth muscle cells (CASMCs). The LDL- and LPA-induced DNA synthesis was markedly inhibited by the LPA receptor antagonist Ki16425, pertussis toxin, small interfering RNAs targeted for LPA1 receptors, and a potent calcineurin inhibitor cyclosporine A. It has been reported that LDL and LPA induced a migration response in a manner sensitive to Ki16425, pertussis toxin, and a LPA1 receptor specific small interfering RNA. However, cyclosporine A was ineffective in inhibiting the migration response. Instead, an epidermal growth factor (EGF) receptor tyrosine kinase inhibitor markedly suppressed the migration response to LDL and LPA without having any significant effect on DNA synthesis. Thus, the LDL induced stimulation of DNA synthesis and migration in CASMCs is mediated by its component LPA through LPA1 receptors and G(i/o)-proteins. Ca2+/calcineurin pathways and transactivation of EGF receptors mediate LPA1-receptor-induced DNA synthesis and migration, respectively. PMID- 19563738 TI - Modelling on the effect of diffusive and convective substrate transport for biofilm. AB - A one-dimensional biofilm model was developed based on the basic principle of conservation of mass. Three simple, generic processes were combined in the model which includes microbial growth, diffusive and convective mass transport. The final model could generate a quantitative description of the relationship between the microbial growth and the consumption of substrate (oxygen) within the fixed biofilm thickness. Mass transfer resistance contributes large influence on the substrates and microbial concentration across the biofilm thickness due to the effect of biofilm structure. PMID- 19563737 TI - Endothelin receptor blockade does not affect blood pressure or angiotensin II levels in CYP1A1-Ren-2 transgenic rats with acutely induced hypertension. AB - We found previously that selective blockade of endothelin ETA receptors is superior to nonselective ET(A)/ET(B) in attenuating hypertension and survival rate in Ren-2 transgenic rats (TGR). In the present pilot study, we were interested in whether similar effects will be found in TGR with inducible malignant hypertension (iTGR; official strain name Cyp1A1-Ren-2rats), which were derived from the original Ren-2 transgenic rat strain. Studies were performed in three-month old male iTGR. Treatment with either bosentan, a non-selective ET(A)/ET(B), or with atrasentan, a selective ET(A) receptor blocker, was started on day 2 of the experiment. Feeding with indole-3-carbinole (13C; 03% in rat chow), a natural xenobiotic which activates the Cyplal promoter of the mouse Ren 2 gene, began on day 3 and lasted for 4 days until day 6. Systolic BP, body weight, plasma ANG II and tissue ANG II and ET-1 concentrations were determined daily. Severe hypertension developed as early as 1 day after beginning of 13C feeding which was accompanied by a significant reduction in body weight and by increases in plasma and tissue ANG II and left ventricle ET-1 concentrations. Atrasentan or bosentan had no effects on the rise in BP or plasma and tissue ANG II concentrations but prevented the rise in heart ventricle ET-1 concentration. Our data show that blockade of the ET system does not prevent or attenuate the rapid development of severe hypertension in iTGR; a long-term protective effect of ET blockade on cardiac (and renal) damage, however, cannot be excluded and awaits further investigations. PMID- 19563739 TI - A note on heroin epidemics. AB - We show that the steady states of the White and Comiskey [E. White, C. Comiskey, Heroin epidemics, treatment and ODE modelling, Math. Biosci. 208 (2007) 312-324.] model of heroin epidemics are stable. PMID- 19563741 TI - A macroscopic model of traffic jams in axons. AB - The purpose of this paper is to develop a minimal macroscopic model capable of explaining the formation of traffic jams in fast axonal transport. The model accounts for the decrease of the number density of positively (and negatively) oriented microtubules near the location of the traffic jam due to formation of microtubule swirls; the model also accounts for the reduction of the effective velocity of organelle transport in the traffic jam region due to organelles falling off microtubule tracks more often in the swirl region. The model is based on molecular-motor-assisted transport equations and the hydrodynamic model of traffic jams in highway traffic. Parametric analyses of the model's predictions for various values of viscosity of the traffic flow, variance of the velocity distribution, diffusivity of microtubule-bound and free organelles, rate constants for binding to and detachment from microtubules, relaxation time, and average motor velocities of the retrograde and anterograde transport, are carried out. PMID- 19563742 TI - Threshold dynamics in a time-delayed epidemic model with dispersal. AB - The global dynamics of a time-delayed model with population dispersal between two patches is investigated. For a general class of birth functions, persistence theory is applied to prove that a disease is persistent when the basic reproduction number is greater than one. It is also shown that the disease will die out if the basic reproduction number is less than one, provided that the initial size of the infected population is relatively small. Numerical simulations are presented using some typical birth functions from biological literature to illustrate the main ideas and the relevance of dispersal. PMID- 19563743 TI - An epidemic model with infector and exposure dependent severity. AB - A stochastic epidemic model allowing for both mildly and severely infectious individuals is defined, where an individual can become severely infectious directly upon infection or if additionally exposed to infection. It is shown that, assuming a large community, the initial phase of the epidemic may be approximated by a suitable branching process and that the main part of an epidemic that becomes established admits a law of large numbers and a central limit theorem, leading to a normal approximation for the final outcome of such an epidemic. Effects of vaccination prior to an outbreak are studied and the critical vaccination coverage, above which only small outbreaks can occur, is derived. The results are illustrated by simulations that demonstrate that the branching process and normal approximations work well for finite communities, and by numerical examples showing that the final outcome may be close to discontinuous in certain model parameters and that the fraction mildly infected may actually increase as an effect of vaccination. PMID- 19563744 TI - Modelling local and global effects on the risk of contracting Tuberculosis using stochastic Markov-chain models. AB - For some diseases, the transmission of infection can cause spatial clustering of disease cases. This clustering has an impact on how one estimates the rate of the spread of the disease and on the design of control strategies. It is, however, difficult to assess such clustering, (local effects on transmission), using traditional statistical methods. A stochastic Markov-chain model that takes into account possible local or more dispersed global effects on the risk of contracting disease is introduced in the context of the transmission dynamics of tuberculosis. The model is used to analyse TB notifications collected in the Asembo and Gem Divisions of Nyanza Province in western Kenya by the Kenya Ministry of Health/National Leprosy and Tuberculosis Program and the Centers for Disease Control and Prevention. The model shows evidence of a pronounced local effect that is significantly greater than the global effect. We discuss a number of variations of the model which identify how this local effect depends on factors such as age and gender. Zoning/clustering of villages is used to identify the influence that zone size has on the model's ability to distinguish local and global effects. An important possible use of the model is in the design of a community randomised trial where geographical clusters of people are divided into two groups and the effectiveness of an intervention policy is assessed by applying it to one group but not the other. Here the model can be used to take the effect of case clustering into consideration in calculating the minimum difference in an outcome variable (e.g. disease prevalence) that can be detected with statistical significance. It thereby gauges the potential effectiveness of such a trial. Such a possible application is illustrated with the given time/spatial TB data set. PMID- 19563745 TI - Seeing green in conference season. AB - Researchers and scientific organizations are becoming aware of the greenhouse-gas emissions and waste associated with attending scientific conferences. Fledgling efforts are now underway to address this problem by offering carbon offsets, recycling at conferences, reducing conference travel, or replacing meetings with teleconferences. PMID- 19563746 TI - Common ancestry of the CENP-A chaperones Scm3 and HJURP. PMID- 19563747 TI - GABA keeps up an appetite for life. AB - In the arcuate nucleus of the hypothalamus, neurons that produce the neuropeptides NPY and AgRP play a vital role in the maintenance of energy homeostasis. In this issue, Wu et al. (2009) show that these neurons modulate feeding behavior in mice by providing GABAergic input to the parabrachial nucleus in the brainstem. PMID- 19563748 TI - Ironing out a midlife crisis. AB - There is a strong correlation between age, genomic instability, and the development of cancer. Working in yeast, Veatch et al. (2009) now propose that defects in the biogenesis of iron-sulfur clusters arising as a consequence of mitochondrial dysfunction contribute to the increase in genomic instability as cells age. PMID- 19563749 TI - The juxtamembrane region of EGFR takes center stage. AB - The activation process for the epidermal growth factor receptor (EGFR) involves formation of an asymmetric dimer of the tyrosine kinase domains. Jura et al. (2009) in this issue and Brewer et al. (2009) in Molecular Cell now demonstrate that the juxtamembrane region of EGFR plays a crucial role in stabilizing this dimer. PMID- 19563750 TI - Closing in on mechanisms of tissue morphogenesis. AB - It remains largely unknown how large-scale tissue movements during development emerge from the interplay of different tensile forces associated with actomyosin networks. Solon et al. (2009) now report that a ratchet-like mechanism drives the movement of epithelial sheets during dorsal closure in embryos of the fruit fly Drosophila. PMID- 19563751 TI - Fly stem cell research gets infectious. AB - To maintain tissue homeostasis, stem cells need to increase their proliferation rate to repair tissue damage caused by stress or infection. In this issue, Jiang et al. (2009) describe a regulatory feedback mechanism involving the Jak/Stat signaling pathway that enables stem cells of the fly midgut to accomplish this task. PMID- 19563752 TI - Plant asymmetric cell division, vive la difference! AB - Although little is known about how asymmetric cell division in plants is regulated, recent discoveries provide a starting point for exploring the mechanisms underlying this process. These studies reveal parallels with asymmetric division in yeast and animals, but also point to regulated cell expansion as a new mechanism of asymmetric division in plants. PMID- 19563753 TI - CTCF: master weaver of the genome. AB - CTCF is a highly conserved zinc finger protein implicated in diverse regulatory functions, including transcriptional activation/repression, insulation, imprinting, and X chromosome inactivation. Here we re-evaluate data supporting these roles in the context of mechanistic insights provided by recent genome-wide studies and highlight evidence for CTCF-mediated intra- and interchromosomal contacts at several developmentally regulated genomic loci. These analyses support a primary role for CTCF in the global organization of chromatin architecture and suggest that CTCF may be a heritable component of an epigenetic system regulating the interplay between DNA methylation, higher-order chromatin structure, and lineage-specific gene expression. PMID- 19563754 TI - Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. AB - LDL delivers cholesterol to lysosomes by receptor-mediated endocytosis. Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. NPC2 binds cholesterol with its isooctyl side chain buried and its 3beta-hydroxyl exposed. Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. NPC1(NTD) binds cholesterol in an orientation opposite to NPC2: 3beta-hydroxyl buried and isooctyl side chain exposed. Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. We propose a working model wherein after lysosomal hydrolysis of LDL cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. PMID- 19563755 TI - Loss of GABAergic signaling by AgRP neurons to the parabrachial nucleus leads to starvation. AB - Neurons in the arcuate nucleus that produce AgRP, NPY, and GABA (AgRP neurons) promote feeding. Ablation of AgRP neurons in adult mice results in Fos activation in postsynaptic neurons and starvation. Loss of GABA is implicated in starvation because chronic subcutaneous delivery of bretazenil (a GABA(A) receptor partial agonist) suppresses Fos activation and maintains feeding during ablation of AgRP neurons. Moreover, under these conditions, direct delivery of bretazenil into the parabrachial nucleus (PBN), a direct target of AgRP neurons that also relays gustatory and visceral sensory information, is sufficient to maintain feeding. Conversely, inactivation of GABA biosynthesis in the ARC or blockade of GABA(A) receptors in the PBN of mice promote anorexia. We suggest that activation of the PBN by AgRP neuron ablation or gastrointestinal malaise inhibits feeding. Chronic delivery of bretazenil during loss of AgRP neurons provides time to establish compensatory mechanisms that eventually allow mice to eat. PMID- 19563756 TI - Abnormal behavior in a chromosome-engineered mouse model for human 15q11-13 duplication seen in autism. AB - Substantial evidence suggests that chromosomal abnormalities contribute to the risk of autism. The duplication of human chromosome 15q11-13 is known to be the most frequent cytogenetic abnormality in autism. We have modeled this genetic change in mice by using chromosome engineering to generate a 6.3 Mb duplication of the conserved linkage group on mouse chromosome 7. Mice with a paternal duplication display poor social interaction, behavioral inflexibility, abnormal ultrasonic vocalizations, and correlates of anxiety. An increased MBII52 snoRNA within the duplicated region, affecting the serotonin 2c receptor (5-HT2cR), correlates with altered intracellular Ca(2+) responses elicited by a 5-HT2cR agonist in neurons of mice with a paternal duplication. This chromosome engineered mouse model for autism seems to replicate various aspects of human autistic phenotypes and validates the relevance of the human chromosome abnormality. This model will facilitate forward genetics of developmental brain disorders and serve as an invaluable tool for therapeutic development. PMID- 19563757 TI - Mitochondrial dysfunction leads to nuclear genome instability via an iron-sulfur cluster defect. AB - Mutations and deletions in the mitochondrial genome (mtDNA), as well as instability of the nuclear genome, are involved in multiple human diseases. Here, we report that in Saccharomyces cerevisiae, loss of mtDNA leads to nuclear genome instability, through a process of cell-cycle arrest and selection we define as a cellular crisis. This crisis is not mediated by the absence of respiration, but instead correlates with a reduction in the mitochondrial membrane potential. Analysis of cells undergoing this crisis identified a defect in iron-sulfur cluster (ISC) biogenesis, which requires normal mitochondrial function. We found that downregulation of nonmitochondrial ISC protein biogenesis was sufficient to cause increased genomic instability in cells with intact mitochondrial function. These results suggest mitochondrial dysfunction stimulates nuclear genome instability by inhibiting the production of ISC-containing protein(s), which are required for maintenance of nuclear genome integrity. For a video summary of this article, see the PaperFlick file available with the online Supplemental Data. PMID- 19563758 TI - Genomic antagonism between retinoic acid and estrogen signaling in breast cancer. AB - Retinoic acid (RA) triggers antiproliferative effects in tumor cells, and therefore RA and its synthetic analogs have great potential as anticarcinogenic agents. Retinoic acid receptors (RARs) mediate RA effects by directly regulating gene expression. To define the genetic network regulated by RARs in breast cancer, we identified RAR genomic targets using chromatin immunoprecipitation and expression analysis. We found that RAR binding throughout the genome is highly coincident with estrogen receptor alpha (ERalpha) binding, resulting in a widespread crosstalk of RA and estrogen signaling to antagonistically regulate breast cancer-associated genes. ERalpha- and RAR-binding sites appear to be coevolved on a large scale throughout the human genome, often resulting in competitive binding activity at nearby or overlapping cis-regulatory elements. The highly coordinated intersection between these two critical nuclear hormone receptor signaling pathways provides a global mechanism for balancing gene expression output via local regulatory interactions dispersed throughout the genome. PMID- 19563760 TI - Mechanism for activation of the EGF receptor catalytic domain by the juxtamembrane segment. AB - Signaling by the epidermal growth factor receptor requires an allosteric interaction between the kinase domains of two receptors, whereby one activates the other. We show that the intracellular juxtamembrane segment of the receptor, known to potentiate kinase activity, is able to dimerize the kinase domains. The C-terminal half of the juxtamembrane segment latches the activated kinase domain to the activator, and the N-terminal half of this segment further potentiates dimerization, most likely by forming an antiparallel helical dimer that engages the transmembrane helices of the activated receptor. Our data are consistent with a mechanism in which the extracellular domains block the intrinsic ability of the transmembrane and cytoplasmic domains to dimerize and activate, with ligand binding releasing this block. The formation of the activating juxtamembrane latch is prevented by the C-terminal tails in a structure of an inactive kinase domain dimer, suggesting how alternative dimers can prevent ligand-independent activation. PMID- 19563759 TI - A synthetic genetic edge detection program. AB - Edge detection is a signal processing algorithm common in artificial intelligence and image recognition programs. We have constructed a genetically encoded edge detection algorithm that programs an isogenic community of E. coli to sense an image of light, communicate to identify the light-dark edges, and visually present the result of the computation. The algorithm is implemented using multiple genetic circuits. An engineered light sensor enables cells to distinguish between light and dark regions. In the dark, cells produce a diffusible chemical signal that diffuses into light regions. Genetic logic gates are used so that only cells that sense light and the diffusible signal produce a positive output. A mathematical model constructed from first principles and parameterized with experimental measurements of the component circuits predicts the performance of the complete program. Quantitatively accurate models will facilitate the engineering of more complex biological behaviors and inform bottom up studies of natural genetic regulatory networks. PMID- 19563761 TI - Exocytosis of post-Golgi vesicles is regulated by components of the endocytic machinery. AB - Post-Golgi vesicles target and deliver most biosynthetic cargoes to the cell surface. However, the molecules and mechanisms involved in fusion of these vesicles are not well understood. We have employed a system to simultaneously monitor release of luminal and membrane biosynthetic cargoes from individual post Golgi vesicles. Exocytosis of these vesicles is not calcium triggered. Release of luminal cargo can be accompanied by complete, partial, or no release of membrane cargo. Partial and no release of membrane cargo of a fusing vesicle are fates associated with kiss-and-run exocytosis, and we find that these are the predominant mode of post-Golgi vesicle exocytosis. Partial cargo release by post Golgi vesicles occurs because of premature closure of the fusion pore and is modulated by the activity of clathrin, actin, and dynamin. Our results demonstrate that these components of the endocytic machinery modulate the nature and extent of biosynthetic cargo delivery by post-Golgi vesicles at the cell membrane. PMID- 19563762 TI - Pulsed forces timed by a ratchet-like mechanism drive directed tissue movement during dorsal closure. AB - Dorsal closure is a tissue-modeling process in the developing Drosophila embryo during which an epidermal opening is closed. It begins with the appearance of a supracellular actin cable that surrounds the opening and provides a contractile force. Amnioserosa cells that fill the opening produce an additional critical force pulling on the surrounding epidermal tissue. We show that this force is not gradual but pulsed and occurs long before dorsal closure starts. Quantitative analysis, combined with laser cutting experiments and simulations, reveals that tension-based dynamics and cell coupling control the force pulses. These constitutively pull the surrounding epidermal tissue dorsally, but the displacement is initially transient. It is translated into dorsal-ward movement only with the help of the actin cable, which acts like a ratchet, counteracting ventral-ward epidermis relaxation after force pulses. Our work uncovers a sophisticated mechanism of cooperative force generation between two major forces driving morphogenesis. PMID- 19563763 TI - Cytokine/Jak/Stat signaling mediates regeneration and homeostasis in the Drosophila midgut. AB - Cells in intestinal epithelia turn over rapidly due to damage from digestion and toxins produced by the enteric microbiota. Gut homeostasis is maintained by intestinal stem cells (ISCs) that divide to replenish the intestinal epithelium, but little is known about how ISC division and differentiation are coordinated with epithelial cell loss. We show here that when enterocytes (ECs) in the Drosophila midgut are subjected to apoptosis, enteric infection, or JNK-mediated stress signaling, they produce cytokines (Upd, Upd2, and Upd3) that activate Jak/Stat signaling in ISCs, promoting their rapid division. Upd/Jak/Stat activity also promotes progenitor cell differentiation, in part by stimulating Delta/Notch signaling, and is required for differentiation in both normal and regenerating midguts. Hence, cytokine-mediated feedback enables stem cells to replace spent progeny as they are lost, thereby establishing gut homeostasis. PMID- 19563764 TI - SnapShot: F Box Proteins II. PMID- 19563765 TI - Using nonfluorescent Forster resonance energy transfer acceptors in protein binding studies. AB - The purpose of this article is to highlight the versatility of nonfluorescent Forster resonance energy transfer (FRET) acceptors in determination of protein equilibrium dissociation constants and kinetic rates. Using a nonfluorescent acceptor eliminates the necessity to spectrally isolate the donor fluorescence when performing binding titrations covering a broad range of reagent concentrations. Moreover, random distribution of the donor and acceptor chromophores on the surface of proteins increases the probability of FRET occurring on their interaction. Three high-affinity antibodies are presented in this study as characteristic protein systems. Monoclonal antibody (mAb) 106.3 binds brain natriuretic peptide (BNP)5-13(C10A) and full-length BNP1-32 with the dissociation constants 0.26+/-0.01 and 0.05+/-0.02 nM, respectively, which was confirmed by kinetic measurements. For anti-hCG (human chorionic gonadotropin) mAb 8F11, studied at two incorporation ratios (IRs=1.9 and 3.8) of the nonfluorescent FRET acceptor, K(D) values of 0.04+/-0.02 and 0.059( 0.004)(+0.006) nM, respectively, were obtained. Likewise, the binding of goat anti-hamster immunoglobulin G (IgG) antibody was not affected by conjugation and yielded K(D) values of 1.26+/-0.04, 1.25+/-0.05, and 1.14+/-0.04 nM at IRs of 1.7, 4.7, and 8.1, respectively. We conclude that this FRET-based method offers high sensitivity, practical simplicity, and versatility in protein binding studies. PMID- 19563766 TI - Identification of heme propionate vibrational modes in the resonance Raman spectra of cytochrome c oxidase. AB - The propionate groups of heme a and a(3) in cytochrome c oxidase (CcO) have been postulated to mediate both the electron and proton transfer within the enzyme. To establish structural markers for the propionate groups, their associated vibrational modes were identified in the resonance Raman spectra of CcO from bovine (bCcO) and Rhodobacter sphaeroides (RsCcO). The distinction between the modes from the propionates of heme a and heme a(3), as well as those from the propionates on the pyrrole rings A and D in each heme, was made on the basis of H2O-D2O isotope substitution experiments combined with wavelength-selective resonance enhancement (for bCcO) or mutagenesis studies (for RsCcO). PMID- 19563767 TI - Surface-enhanced Raman spectroscopy combined with atomic force microscopy for ultrasensitive detection of thrombin. AB - We have developed an ultrasensitive analytical method based on surface-enhanced Raman spectroscopy (SERS) exploiting a Raman probe covalently bound to gold nanoparticles. The biological marker to be detected was adsorbed on functionalized gold nanoparticles. The capture of these nanoparticles via a biorecognition process between the marker and the immobilized receptor was demonstrated by atomic force microscopy (AFM) imaging. The vibrational fingerprints of the Raman probe on the capture substrate were followed to reveal the presence of the biological marker. The method, which was applied to reveal thrombin captured on a substrate containing antithrombin and heparin, resulted in the ability to detect marker concentrations down to the picomolar (pM) level. PMID- 19563768 TI - A colorimetric assay for sulfiredoxin activity using inorganic phosphate measurement. AB - 2-Cys peroxiredoxin (Prx) is the major subgroup of a family of Prx enzymes that reduce peroxide molecules such as hydrogen peroxide (H(2)O(2)). 2-Cys Prxs are inactivated when their active site cysteine residue is hyperoxidized to sulfinic acid. Sulfiredoxin (Srx) is an enzyme that catalyzes reduction of hyperoxidized 2 Cys Prxs in the presence of ATP, Mg(2+), and thiol equivalent. Therefore, Srx activity is crucial for cellular function of 2-Cys Prxs. The method currently available for the determination of Srx activity relies on immunoblot detection using antibodies to hyperoxidized enzymes. Here we introduce a simple quantitative assay for Srx activity based on the colorimetric determination of inorganic phosphate released in Srx-dependent reduction of hyperoxidized Prx using the malachite green. The colorimetric assay was used for high-throughput screening of 25,000 chemicals to find Srx inhibitors. PMID- 19563769 TI - Use of mass spectrometry to probe the nucleophilicity of cysteinyl residues of prolyl hydroxylase domain 2. AB - Prolyl hydroxylase domain 2 (PHD2) plays an important role in hypoxic sensing in humans. Here we report studies on the reactivity of cysteinyl residues of the catalytic domain of PHD2 using an approach in which nondenaturing electrospray ionization-mass spectrometry (ESI-MS) analyses were combined with the use of a thiol library and residue substitution. Among the seven cysteinyl residues of the PHD2 catalytic domain, Cys201 was found to be predominantly modified by thiols or N-ethylmaleimide. Selective modification of Cys201 was further demonstrated with methanethiosulfonate, a spin-labeled probe. The modified PHD2 will be useful in electron paramagnetic resonance studies on PHD2. The results demonstrate the use of a combined library/residue substitution/ESI-MS approach for analyzing residue reactivity. PMID- 19563770 TI - Determination of oxysterols in oxidatively modified low-density lipoprotein by semi-micro high-performance liquid chromatography with electrochemical detection. AB - A simple method for the determination of oxysterols was developed by semi-micro high-performance liquid chromatography with electrochemical detection (semi-micro HPLC-ECD). Semi-micro HPLC-ECD was established using a C30 microbore column, acetonitrile containing 50 mmol/L LiClO(4) as a mobile phase, and an applied potential at +2.8V versus Ag/AgCl. The current peak height was linearly related to the amount of sterol injected from 12.5 to 250 pmol (r>0.999) with a relative standard deviation (RSD) of less than 2.9% (n=6). This method was applied to the determination of seven oxysterols in oxidatively modified low-density lipoprotein (Ox-LDL). Oxysterols were determined with a recovery of more than 78.0% and an RSD of less than 2.9% (n=6) except for 7-ketocholesterol. 7-Ketocholesterol was determined as a sum of intact 7-ketocholesterol and its degradation product on saponification, cholesta-3,5-dien-7-one, with a recovery of 98.0% and an RSD of 2.5% (n=6). From these results, the current method enabled the simultaneous determination of seven oxysterols without any derivatization, providing a useful tool for the assessment of oxysterol contents in Ox-LDL. PMID- 19563771 TI - Quantitative isotopic 13C nuclear magnetic resonance at natural abundance to probe enzyme reaction mechanisms via site-specific isotope fractionation: the case of the chain-shortening reaction for the bioconversion of ferulic acid to vanillin. AB - Isotope fractionation is a powerful technique by which to probe the reaction mechanism of enzymes. The effect of a heavy isotope on the reaction energetics can be used to predict transition state architecture and reaction mechanism. In order to examine simultaneously the isotope fractionation in (13)C at multiple sites within the substrate and product molecules without any need for site selective isotope enrichment, a technique exploiting quantitative isotopic nuclear magnetic resonance (NMR) spectrometry at natural abundance (NAQ-NMR) has been developed. Here we report the first application of this technique to the study of an enzyme-catalyzed reaction, the bioconversion of ferulic acid to vanillin in cultures of Streptomyces setonii. We were able to show that the NAQ NMR methodology is sufficiently precise and robust to measure the isotope shifts in the (13)C/(12)C ratios in both substrate and product of this biotransformation, thereby permitting meaningful data to be obtained even at carbon positions that take part only indirectly in the reaction and show only secondary isotope fractionation. The results obtained provide direct evidence in support of the current hypothesis for the reaction mechanism of the enzyme hydroxycinnamoyl-CoA hydratase/lyase, notably the proposed involvement of the quinone methide enolate of feruloyl-CoA as intermediate in the catalytic pathway. PMID- 19563772 TI - Cytochrome P450-dependent metabolism of PCB52 in the nematode Caenorhabditis elegans. AB - There are 75 full length cytochrome P450 (CYP) genes known in the genome of the nematode Caenorhabditis elegans. The individual biological functions of the vast majority are mostly as yet unknown. Here the impact of cytochrome P450 isoforms on the metabolism of PCB52, an ortho-substituted, non-coplanar 2,2',5,5' tetrachlorbiphenyl, as a model PCB of these worldwide distributed pollutants is investigated. Organic extracts, isolated from treated worms and analyzed by GC/MS, contained two obvious PCB52-derived products which have been identified as C3-, C4- and/or C6-hydroxy-PCB52. Moreover, these hydroxylase reactions strictly required the functional expression of the NADPH-dependent cytochrome P450 reductase (CPR) encoding emb-8 gene, which was recently shown to be essential also for several other cytochrome P450-dependent enzymatic reactions. Multiple and subsequent single RNAi-gene silencing experiments, as well as the use of cyp mutant strains, identified members of the CYP-14A subfamily and CYP-34A6 as the major isoforms contributing to PCB52 metabolism in C. elegans. In the gene silenced worms and mutants, the reduction in formation of hydroxylated products ranged from 55% to 78%. These results demonstrate for the first time that C. elegans shares with mammals the capacity to produce CYP-dependent PCB metabolites and may thus facilitate future studies on biotransformation. PMID- 19563774 TI - The calcitonin/calcitonin gene related peptide-alpha gene is not required for 1alpha,25-dihydroxyvitamin D3-mediated suppression of experimental autoimmune encephalomyelitis. AB - The active form of vitamin D, 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), can suppress disease in the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis. Calcium appears to be a critical component of 1,25(OH)(2)D(3)-mediated suppression of EAE, as complete disease prevention only occurs with a concomitant increase in serum calcium levels. Calcitonin (CT) is a peptide hormone released in response to acute increases in serum calcium, which led us to explore its importance in 1,25(OH)(2)D(3)-mediated suppression of EAE. Previously, we discovered that co-administration of pharmacological doses of CT enhanced the suppressive effect of 1,25(OH)(2)D(3) on EAE, suggesting CT may play a role in 1,25(OH)(2)D(3)-mediated suppression of EAE. To determine the importance of CT in EAE we have utilized a mouse strain in which the gene encoding CT and its alternative splice product, calcitonin gene related peptide alpha (CGRP), have been deleted. Deletion of the CT/CGRP gene had no effect on EAE progression. Furthermore, treatment with 1,25(OH)(2)D(3) suppressed EAE in CT/CGRP knock-out mice equal to that in wild type mice. Therefore, we conclude that CT is not necessary for 1,25(OH)(2)D(3)-mediated suppression of EAE. PMID- 19563775 TI - Overexpression of Rac1 in leukemia patients and its role in leukemia cell migration and growth. AB - Rac1 belongs to the Rho family that act as critical mediators of signaling pathways controlling cell migration and proliferation and contributes to the interactions of hematopoietic stem cells with their microenvironment. Alteration of Rac1 might result in unbalanced interactions and ultimately lead to leukemogenesis. In this study, we analyze the expression of Rac1 protein in leukemia patients and determine its role in the abnormal behaviours of leukemic cells. Rac1 protein is overexpressed in primary acute myeloid leukemia cells as compared to normal bone marrow mononuclear cells. siRNA-mediated silencing of Rac1 in leukemia cell lines induced inhibition of cell migration, proliferation, and colony formation. Additionally, blocking Rac1 activity by an inhibitor of Rac1-GTPase, NSC23766, suppressed cell migration and growth. We conclude that overexpression of Rac1 contributes to the accelerated migration and high proliferation potential of leukemia cells, which could be implicated in leukemia development and progression. PMID- 19563773 TI - Inhibition of human folylpolyglutamate synthetase by diastereomeric phosphinic acid mimics of the tetrahedral intermediate. AB - Phosphorus-containing pseudopeptides, racemic at the C-terminal alpha-carbon, are potent mechanism-based inhibitors of folylpolyglutamate synthetase (FPGS). They are mimics of the tetrahedral intermediate postulated to form during FPGS catalyzed biosynthesis of poly(gamma-l-glutamates). In the present paper, the FPGS inhibitory activity of each diastereomer coupled to three heterocycles is reported. The high R(f) pseudopeptide containing the 5,10 dideazatetrahydropteroyl (DDAH(4)Pte) heterocycle is most potent (K(is) = 1.7 nM). While the heterocyclic portion affects absolute FPGS inhibitory potency, the high R(f) species is more potent in each pair containing the same heterocycle. This species presumably has the same stereochemistry as the natural folate polyglutamate, i.e., (l-Glu-gamma-l-Glu). Unexpectedly, the low R(f) (presumed l Glu-gamma-d-Glu) species are only slightly less potent (<30-fold) than their diastereomers. Further study of this phenomenon comparing l-Glu-gamma-l-Glu and l Glu-gamma-d-Glu dipeptide-containing FPGS substrates shows that <1% contamination of commercial d-Glu precursors by l-Glu may give misleading information if l-Glu gamma-l-Glu substrates have low K(m) values. PMID- 19563776 TI - Activity-dependent regulation of HCN1 protein in cortical neurons. AB - Homeostasis of neuronal activity is crucial to neuronal physiology. In dendrites, hyperpolarization-activated cyclic nucleotide-gated channel (HCN) 1 is considered to play critical roles in this process. While electrophysiological studies have demonstrated the dynamic modulation of I(h) current mediated by HCN1 proteins, little is known about the underlying molecular and cellular mechanisms. In this study, we utilized cortical cultured neurons and biochemical methods to identify molecular and cellular mechanisms that mediate the physiological regulation of HCN1 channel functions in cortical neurons. Pharmacological manipulations of neuronal activity resulted in changes in the expression level of HCN1. In addition, the surface expression of HCN1 was dynamically regulated by neuronal activity. Both of these changes led to functional modulations of HCN1 channels. Our study suggests that coordinated changes in protein expression and surface expression of HCN1 serve as the key regulatory mechanisms controlling the function of endogenous HCN1 protein in cortical neurons. PMID- 19563777 TI - Aldo-keto reductase family 1 B10 protein detoxifies dietary and lipid-derived alpha, beta-unsaturated carbonyls at physiological levels. AB - Alpha, beta-unsaturated carbonyls are highly reactive mutagens and carcinogens to which humans are exposed on a daily basis. This study demonstrates that aldo-keto reductase family 1 member B10 (AKR1B10) is a critical protein in detoxifying dietary and lipid-derived unsaturated carbonyls. Purified AKR1B10 recombinant protein efficiently catalyzed the reduction to less toxic alcohol forms of crotonaldehyde at 0.90 microM, 4-hydroxynonenal (HNE) at 0.10 microM, trans-2 hexanal at 0.10 microM, and trans-2,4-hexadienal at 0.05 microM, the concentrations at or lower than physiological exposures. Ectopically expressed AKR1B10 in 293T cells eliminated immediately HNE at 1 (subtoxic) or 5 microM (toxic) by converting to 1,4-dihydroxynonene, protecting the cells from HNE toxicity. AKR1B10 protein also showed strong enzymatic activity toward glutathione-conjugated carbonyls. Taken together, our study results suggest that AKR1B10 specifically expressed in the intestine is physiologically important in protecting the host cell against dietary and lipid-derived cytotoxic carbonyls. PMID- 19563778 TI - Daxx positively modulates beta-catenin/TCF4-mediated transcriptional potential. AB - Constitutive activation of the transcription factor TCF4 activity by mutated APC or beta-catenin contributes to cell neoplastic transformation. While numerous proteins were identified to activate TCF4-dependent activity via beta-catenin interaction, little is known about factors directly acting on TCF4. Here we report that Daxx binds to TCF4 and potentiates beta-catenin/TCF4-mediated transcriptional activation and target gene expression. Binding studies revealed that Daxx-TCF4 interaction is through the C-terminal domain of Daxx and TCF4 segment containing amino acid residue 269-327. Alteration of Daxx levels in cells by overexpression or RNA interference resulted in an increase or decrease of the beta-catenin/TCF4-dependent transactivation activity and target gene expression, respectively. Furthermore, TCF4-(269-327) segment acts as a dominantly negative mutant by blocking Daxx-TCF4 interaction and TCF4-mediated transactivation potential. Together, our results suggest that Daxx functions as a positive coregulator in modulating the beta-catenin/TCF4-dependent transcriptional potential via TCF4 interaction. PMID- 19563779 TI - Forkhead transcription factor FoxO1 inhibits insulin- and transforming growth factor-beta-stimulated plasminogen activator inhibitor-1 expression. AB - Elevated levels of plasminogen activator inhibitor-1 (PAI-1) are considered a risk factor for chronic liver disease in patients with hyperinsulinemia. Insulin increases the expression of PAI-1, and inactivates the forkhead box-containing protein FoxO1. We were interested in whether the inactivation of FoxO1 is involved in the activation of PAI-1 expression under conditions of insulin stimulation. Here, we examined whether adenoviral-mediated expression of a constitutively active form of FoxO1 (Ad-CA-FoxO1) inhibited insulin-stimulated PAI-1 expression in human HepG2 hepatocellular liver carcinoma cells and mouse AML12 hepatocytes. Treatment of cells with insulin increased PAI-1 gene expression, and this effect was abolished by Ad-CA-FoxO1. Insulin also increased the transforming growth factor (TGF)-beta-induced expression of PAI-1 mRNA, and Ad-CA-FoxO1 inhibited this effect. Transient transfection assays using a reporter gene under the control of the PAI-1 promoter revealed that CA-FoxO1 inhibits Smad3-stimulated PAI-1 promoter activity. Taken together, our results indicate that FoxO1 inhibits PAI-1 expression through the inhibition of TGF-beta/Smad mediated signaling pathways. Our data also suggest that in the hyperinsulinemic state, FoxO1 is inactivated by increased levels of insulin, and does not function as an inhibitor of TGF-beta-induced PAI-1 expression. PMID- 19563780 TI - PKCdelta mediates Nrf2-dependent protection of neuronal cells from NO-induced apoptosis. AB - A chemical inhibitor library of 84 compounds was screened to investigate the signaling pathway(s) leading to activation of Nrf2 in response to nitric oxide (NO). We identified the protein kinase C delta (PKCdelta) inhibitor rottlerin as the only compound that reduced NO-induced ARE-luciferase reporter activity and diminished NO-induced up-regulation of two Nrf2/ARE-regulated proteins - NAD(P)H:quinone oxidoreductase-1 (NQO1) and hemeoxygenase-1 (HO-1) in SH-Sy5y cells. Rottlerin also sensitized neuroblastoma cells and mouse primary cortical neurons to NO-induced apoptosis. Stable over-expression of PKCdelta augmented NO induced, ARE-dependent gene expression of HO-1 in SH-Sy5y cells, which were more protected from NO killing. Conversely, NO-induced ARE-dependent gene expression was reduced in PKCdelta-knockdown SH-EP cells, which displayed greater sensitivity to apoptosis. PKCdelta(-/-) cortical neurons exhibited increased NO induced apoptosis and less HO-1 mRNA and protein induction compared with wild type neurons. Hence, PKCdelta is an important positive modulator of NO-induced Nrf2/ARE-dependent signaling that counteracts NO-mediated apoptosis in neuronal cells. PMID- 19563781 TI - Non-fibrillar amyloid-beta peptide reduces NMDA-induced neurotoxicity, but not AMPA-induced neurotoxicity. AB - Amyloid-beta peptide (Abeta) is thought to be linked to the pathogenesis of Alzheimer's disease. Recent studies suggest that Abeta has important physiological roles in addition to its pathological roles. We recently demonstrated that Abeta42 protects hippocampal neurons from glutamate-induced neurotoxicity, but the relationship between Abeta42 assemblies and their neuroprotective effects remains largely unknown. In this study, we prepared non fibrillar and fibrillar Abeta42 based on the results of the thioflavin T assay, Western blot analysis, and atomic force microscopy, and examined the effects of non-fibrillar and fibrillar Abeta42 on glutamate-induced neurotoxicity. Non fibrillar Abeta42, but not fibrillar Abeta42, protected hippocampal neurons from glutamate-induced neurotoxicity. Furthermore, non-fibrillar Abeta42 decreased both neurotoxicity and increases in the intracellular Ca(2+) concentration induced by N-methyl-d-aspartate (NMDA), but not by alpha-amino-3-hydrozy-5-methyl 4-isoxazole propionic acid (AMPA). Our results suggest that non-fibrillar Abeta42 protects hippocampal neurons from glutamate-induced neurotoxicity through regulation of the NMDA receptor. PMID- 19563782 TI - Thyroid hormone non-genomically suppresses Src thereby stimulating osteocalcin expression in primary mouse calvarial osteoblasts. AB - To provide further insights into non-genomic action of thyroid hormone (T3), we investigated whether Src is under control of T3 in primary calvarial osteoblasts prepared from neonatal mice. Treatment of the cells with T3 rapidly decreased Src Y416 autophosphorylation, followed by the decrease of phosphorylated extracellular signal-regulated kinases, suggesting that T3 non-genomically suppresses Src activity. Furthermore, this T3 effect was rapid and persistent, and was associated with the increased expression of osteocalcin (OC). To confirm the contribution of Src to the effect of T3 on OC expression, a constitutively active Src (Y527F) was overexpressed in osteoblasts. In such cells, Y416 phosphorylation was markedly increased even in the presence of T3, and T3 dependent expression of OC was markedly attenuated. The present study demonstrates a novel, non-genomic action of T3 in primary mouse osteoblasts, by which T3 suppresses Src thereby stimulating OC expression. PMID- 19563783 TI - Dual knockdown of N-ras and epiregulin synergistically suppressed the growth of human hepatoma cells. AB - Hepatocellular carcinoma (HCC) is a major challenge because of its resistance to conventional cytotoxic chemotherapy and radiotherapy. Multi-targeted therapy might be a new option for HCC treatment. Our previous study showed that N-ras gene was activated in HCC and was inhibited by RNA interference. In the present study, we investigated the alternation of gene expression by microarray in N-Ras siRNA-treated HepG2 cells. The results revealed that the EREG gene, encoding epiregulin, was dramatically up-regulated in response to silence of N-ras. We speculated that the up-regulation of epiregulin was involved in the compensatory mechanism of N-ras knockdown for cell growth. Therefore, we evaluated whether dual silence of N-ras and epiregulin display a greater suppression of cell growth. The results confirmed that dual knockdown of N-ras and epiregulin synergistically inhibited cell growth. Our results also showed that dual knockdown of N-ras and epiregulin significantly induced cell arrest at G0/G1 phase. Furthermore, Western blot assay showed that dual knockdown of N-ras and epiregulin markedly reduced the phosphorylations of ERK1/2, Akt and Rb, and inhibited the expression of cyclin D1. Our findings imply that multi-targeted silence of oncogenes might be an effective treatment for HCC. PMID- 19563784 TI - Altered expression of adenosine A1 and A2A receptors in the carotid body and nucleus tractus solitarius of adult male and female rats following neonatal caffeine treatment. AB - Neonatal caffeine treatment (adenosine receptor antagonist, 15 mg/kg/day, between postnatal days 3 and 12) affects respiratory patterns in adult male but not female rats as shown by an increase in the respiratory frequency in the early phase of response to hypoxia and an increase in the tidal volume in the late phase of response. Here, we tested the hypothesis that these changes are correlated with modified expression of adenosine receptors in the chemoreflex pathway. Carotid bodies, nucleus tractus solitarii, and superior cervical ganglia were collected from 3-month-old male and female rats that were either naive (not manipulated during the neonatal period) or treated with caffeine (NCT) or water (NWT) between postnatal days 3 and 12 by gavage. Western blot analysis was used to assess the expression of adenosine A(1) and A(2A) receptors and tyrosine hydroxylase, the rate-limiting enzyme for dopamine synthesis. In male rats, there was a 37% increase in the level of A(2A) receptor and a 17% decrease in tyrosine hydroxylase in the carotid body of NCT (p<0.001) as compared to NWT rats. In the nucleus tractus solitarius, we found a 13% and 19% decrease in A(1) receptor expression in NWT and NCT rats (p<0.01), respectively, compared to naive rats. In the superior cervical ganglion, there was no change in A(1) receptor, A(2A) receptor, and tyrosine hydroxylase expression. In female rats, the only changes observed were decreases of 12% and 15% in A(1) receptor levels in the nucleus tractus solitarius of NWT and NCT rats (p<0.01), respectively, compared to naive rats. We conclude that NCT induces long-term changes in the adenosine receptor system. These changes may partially explain the modifications of the respiratory pattern induced by NCT in adults. The increased expression of the adenosine A(2A) receptor (specific to male rats), combined with the decreased tyrosine hydroxylase expression in the carotid body, suggests that NCT affects adenosine dopamine interactions regulating chemosensory activity. PMID- 19563785 TI - IGF-1:tetanus toxin fragment C fusion protein improves delivery of IGF-1 to spinal cord but fails to prolong survival of ALS mice. AB - To improve delivery of human insulin-like growth factor-1 (hIGF-1) to brain and spinal cord, we generated a soluble IGF-1:tetanus toxin fragment C fusion protein (IGF-1:TTC) as a secreted product from insect cells. IGF-1:TTC exhibited IGF-1 and TTC activity in vitro; it increased levels of immunoreactive phosphoAkt in treated MCF-7 cells and bound to immobilized ganglioside GT1b. In mice, the fusion protein underwent retrograde transport by spinal cord motor neurons following intramuscular injection, and exhibited both TTC- and IGF-1 activity in the CNS following intrathecal infusion. Analogous to the case with TTC, intrathecal infusion of the fusion protein resulted in substantial levels of IGF 1:TTC in spinal cord tissue extracts. Tissue concentrations of hIGF-1 in lumbar spinal cords of mice infused with IGF-1:TTC were estimated to be approximately 500-fold higher than those in mice treated with unmodified recombinant hIGF-1 (rhIGF-1). Like rhIGF-1, infusion of IGF-1:TTC reduced levels of IGF-1 receptor immunoreactivity in the same extracts. Despite raising levels of exogenous hIGF-1 in spinal cord, intramuscular- or intrathecal administration of IGF-1:TTC had no significant effect on disease progression or survival of high-expressing SOD1(G93A) transgenic mice. IGF-1:TTC may prove to be neuroprotective in other animal models of CNS disease or injury known to be responsive to unmodified IGF 1. PMID- 19563786 TI - Global ischemia-induced modifications in the expression of AMPA receptors and inflammation in rat brain. AB - Alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptors (AMPAR) and inflammatory processes have been related to ischemia-induced damage, but there are few studies addressing their response in different brain areas. Here we compare AMPAR expression after ischemia in several brain areas (hippocampus, cerebral cortex and caudate-putamen) in an attempt to correlate it with their different vulnerabilities. We found outstanding decreases in GluR1 and GluR2 mRNA levels after global ischemia and 48 h reperfusion (I/R) in all the areas studied, however, protein levels maintained in some areas such as CA3, suggesting different post-transcriptional control in different areas of the brain. To characterize the inflammatory response in these areas, we measured the mRNA levels of CD11b/CD18 membrane integrin (a reactive microglia marker), which showed an important but similar up-regulation in all brain areas studied, which was confirmed by immunohistochemistry. We conclude that the down-regulation of AMPAR gene expression following I/R does not explain differences in the vulnerability of different areas. Additionally, our data indicate that the level of inflammation is independent of the vulnerability of the different brain areas and does not explain differences in the AMPAR expression observed in the brain areas studied. PMID- 19563787 TI - Spatiotemporospectral characteristics of scalp ictal EEG in mesial temporal lobe epilepsy with hippocampal sclerosis. AB - The aim of present study was to identify the common propagation pattern of ictal discharges in mesial temporal lobe epilepsy (TLE). Pre-surgical ictal scalp EEG recordings were collected from patients with TLE associated with hippocampal sclerosis. Two types of ictal onset patterns were identified based on spatial and spectral characteristics of initial ictal discharge waveforms: (a) a sustained regular 5- to 9-Hz rhythm with a restricted temporal or subtemporal distribution (type 1); and (b) an irregular 2- to 5-Hz rhythm with a widespread distribution (type 2). Scalp EEG data were decomposed into temporally independent, spatially fixed component by independent component analysis. The identified source activities corresponding to ictal discharges from each seizure were localized by dipole source localization, and dipole sources with similar spatial locations were clustered. To identify the sequence of propagation among component clusters during the progress of seizures, event-related spectral perturbation by wavelet transform was used. Fifty-five seizures (22 seizures in four Type 1 patients and 33 seizures in eight Type 2 patients) in 12 patients were analyzed. Ictal discharges associated with type 1 seizures arose from both the anterior temporal region and basal ganglia, and then spread into medial frontal region. The dominant frequency of ictal rhythm was in the theta range and remained relatively constant until the middle portion of seizures. Type 2 seizures developed bilaterally or predominantly in the ipsilateral medial temporal region, followed by the medial frontal region and basal ganglia. The dominant frequency of ictal activity at the onset of seizure was in the delta range. However, rhythmic theta activities with decreasing tendency ensued rapidly after seizure onset. These findings suggest that TLE associated with hippocampal sclerosis may have preferential propagating patterns according to the type of the ictal onset pattern. PMID- 19563788 TI - Long term high frequency stimulation of STN increases dopamine in the corpus striatum of hemiparkinsonian rhesus monkey. AB - Long term subthalamic nucleus (STN) high frequency stimulation (HFS) can improve most symptoms of Parkinson's disease (PD) patients and decrease the dosage of antiparkinsonian drug such as Madopar. The mechanism of STN HFS for PD still remains elusive. We hypothesize that the level of dopamine (DA) and its metabolites in the corpus striatum is increased after long term STN HFS. The aim of this study was to examine the DA and its metabolites in the extracellular space of corpus striatum in hemiparkinsonian monkeys during long term STN HFS. Four rhesus monkeys were induced to hemiparkinsonian models by injecting 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) through right internal carotid artery. Then two of them were underwent long term right STN HFS for the subsequent microdialysis sessions. Four microdialysis probe cannulas were implanted into bilateral putamen and caudate nucleus respectively. The microdialysis probe was put into the microdialysis probe cannula of bilateral putamen and caudate nucleus. Dialysates of extracellular space in corpus striatum were collected prior to STN HFS, and subsequently 8 h, 1 week, 1 month, 2 months, 8 months and 10 months after STN HFS. The level of DA and its metabolites were determined by high performance liquid chromatography and subthalamic nucleus electrochemical detection (HPLC-ECD). HFS significantly improved PD symptoms of the monkeys. Rotation evoked by apomorphine (APO) disappeared immediately after HFS pulse generator was turned on. The levels of DA and its metabolites in putamen and caudate nucleus of electrode side increased significantly at different time points after stimulation. Long term STN HFS significantly improved symptoms of hemiparkinsonian rhesus monkey, which might be due to the increase of dopamine and/or its metabolites in corpus striatum. PMID- 19563790 TI - Neuroprotective effects of N-stearoyltyrosine on transient global cerebral ischemia in gerbils. AB - N-stearoyltyrosine (NsTyr), a synthesized anandamide (AEA) analogue, was evaluated for the first time in the present study for the neuroprotective effect in gerbils subjected to transient global cerebral ischemia reperfusion (IR). The extent of ischemia injury was assessed behaviorally by measuring neurological functions, passive avoidance test and Morris water maze; and histopathologically by evaluating hippocampal CA1 pyramidal damage. In addition, ischemia-induced apoptosis was examined using the terminal deoxynucleotidyl transferase-mediated UTP nick end labeling (TUNEL) method. Furthermore, in order to understand the mechanism of NsTyr's neuroprotective effect, we examined antioxidative enzymes, such as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH PX), and non-enzymatic scavenger glutathione (GSH) and measured the levels of malondialdehyde (MDA) in hippocampus. The administration of NsTyr led to attenuation of ischemia-induced neural deficits both behaviorally and histopathologically, reduced the level of MDA, significantly increased the activity of antioxidants GSH and GSH-PX, and obviously elevated the activities of SOD and CAT. Our results suggest that NsTyr shows neuroprotective effect on global cerebral IR injury and its neuroprotective effects may be attributed to restraining DNA fragmentation, suppressing the production of free radicals and elevating antioxidant capacity. PMID- 19563789 TI - Fractalkine and CX3CR1 are involved in the migration of intravenously grafted human bone marrow stromal cells toward ischemic brain lesion in rats. AB - Recent research has shown that transplanted bone marrow stromal cells (MSCs) migrate to the injured regions and exert their therapeutic effects in cases of intracranial trauma, stroke, inflammation and degenerative disease. The specific mechanisms involved in their migration to lesions are still to be fully elucidated. In the present study, a rat model of transient middle cerebral artery occlusion (MCAO) was established. At 24 h after reperfusion, human bone marrow stromal cells (hMSCs) were transplanted by intravenous injection to explore the effects of fractalkine/CX3CR1 on the migration of transplanted MSCs to lesions. In vitro study using real-time PCR and western blot revealed that CX3CR1, the only known receptor of fractalkine, was expressed in cultured hMSCs. The expression of fractalkine in the ischemic brain was significantly increased. The directional migration of transplanted hMSCs to the damaged region was observed through detection of green fluorescence protein (GFP). The results indicated the cells were mainly distributed in the ischemic boundary zone with high fractalkine expression. In a further study, lentivirus-mediated RNA interference of CX3CR1 expression was employed. The results of these experiments indicated that CX3CR1 knock-down dramatically decreased the migration of hMSCs to the ischemic brain. The present study suggests that fractalkine and its specific receptor CX3CR1 are involved in the directional migration of transplanted MSCs to the ischemic damaged brain region. PMID- 19563791 TI - Specifications for trueness and precision of a reference measurement system for serum/plasma 25-hydroxyvitamin D analysis. AB - BACKGROUND: The divergence in analytical quality of serum/plasma 25-hydroxy vitamin D analysis calls for defining specifications for a reference measurement system. METHODS: Fundamentally, in a reference measurement system, there should be a relationship between the analytical specifications for higher- (reference) and lower-order (routine) measurements. Therefore, when setting specifications, we started with limits for routine imprecision (CV(rou)) and bias (B(rou)) using 4 models: (1) the misclassifications in diagnosis, (2) biological variation data (reference interval (RI) and monitoring), (3) expert recommendations, and (4) state-of-the-art performance. Then, we used the derived goals to tailor those for reference measurements and certified reference materials (CRMs) for calibration by setting the limits for CV(ref) at 0.5 CV(rou), B(ref) at 0.33 B(rou)(,) max. uncertainty (U(max)) at 0.33 B(ref). RESULTS: The established specifications ranged between CV(rou)or=0.91 at three sites and mean biases of 14%, 25% and 39%. CMIA patient specimen correlations to the Abbott IMx gave R>or=0.94 at 2 sites and mean biases of 5.4% and 6.9%. CONCLUSIONS: CMIA is a precise and sensitive immunoassay method without hematocrit interference. It correlates well to both LC/MS/MS and immunoassay results, but shows an expected positive bias to LC/MS/MS. PMID- 19563795 TI - Comparison of growth factor and cytokine expression in patients with degenerated disc disease and herniated nucleus pulposus. AB - OBJECTIVES: This study was conducted to investigate the expression of cytokines and growth factors in disc specimens obtained from patients with herniated nucleus pulposus (HNP) and degenerated disc disease (DDD). DESIGN AND METHODS: MRI and Western blot analyses were performed to evaluate the levels of disc degeneration and the expression levels of cytokines and growth factors. RESULTS: The levels of TNF-alpha and IL-8 were significantly greater in the DDD group than in the HNP group, but no statistical differences were observed in the expression of IL-1beta, IL-6 and IL-12 between the HNP and DDD groups. In addition, the expression of TGF beta, VEGF and NGF was significantly higher in the DDD group than in the HNP group. CONCLUSION: The greater levels of cytokine and growth factor expression in the DDD group than in the HNP explain why discogenic patients usually have more severe back pain than patients with herniated discs. PMID- 19563796 TI - The apical/basal-polarity determinant Scribble cooperates with the PCP core factor Stbm/Vang and functions as one of its effectors. AB - Most tissues display several features of cellular polarization. Besides the ubiquitous epithelial polarization in the Apical-Basal (A/B) axis, many epithelia (and associated organs) display a Planar Cell Polarization (PCP). Recently, a crosstalk between the PCP and A/B polarity determinants has been suggested, i.e. the activity or stability of the PCP factor Frizzled is regulated by the A/B determinants aPKC and Bazooka in the Drosophila eye. We have systematically investigated genetic and physical interactions between the Drosophila A/B factors and the core PCP component Strabismus (Stbm)/Van Gogh (Vang). The A/B determinant Scribble was found to interact both genetically and physically with Stbm/Vang. We demonstrate that Scribble binds Stbm/Vang through its PDZ domain 3 and that it cooperates with Stbm/Vang in PCP establishment. Our data indicate that Scribble, in addition to its role in A/B polarity, has a distinct requirement in PCP establishment in the Drosophila eye and wing. We define a scribble allele that is largely PCP specific. Our data show that Scribble is part of the Stbm/Vang PCP complex and further suggest that it might act as an effector of Stbm/Vang during PCP establishment. PMID- 19563797 TI - Tbx2 misexpression impairs deployment of second heart field derived progenitor cells to the arterial pole of the embryonic heart. AB - Tbx2 is a member of the T-box family of transcription factors that play important roles during heart development. In the embryonic heart tube, Tbx2 is expressed in non-chamber myocardium (outflow tract and interventricular canal) and has been shown to block chamber formation. We have developed a genetic system to conditionally misexpress Tbx2 in the embryonic mouse heart at early stages of development. We show that Tbx2 expression throughout the myocardium of the heart tube both represses proliferation and impairs secondary heart field (SHF) progenitor cell deployment into the outflow tract (OFT). Repression of proliferation is accompanied by the upregulation of Ndrg2 and downregulation of Ndrg4 expression, both genes believed to be involved in cell growth and proliferation. Impaired deployment of SHF cells from the pharyngeal mesoderm is accompanied by downregulation of the cell adhesion molecules Alcam and N-cadherin in the anterior part of the embryonic heart. Tbx2 misexpression also results in downregulation of Tbx20 within the OFT, indicating complex and region-specific transcriptional cross-regulation between the two T-box genes. PMID- 19563798 TI - Pax6 dosage requirements in iris and ciliary body differentiation. AB - Pax6 is a highly conserved transcription factor that controls the morphogenesis of various organs. Changes in Pax6 dosage have been shown to affect the formation of multiple tissues. PAX6 haploinsufficiency leads to aniridia, a pan-ocular disease primarily characterized by iris hypoplasia. Herein, we employ a modular system that includes null and overexpressed conditional alleles of Pax6. The use of the Tyrp2-Cre line, active in iris and ciliary body (CB) primordium, enabled us to investigate the effect of varying dosages of Pax6 on the development of these ocular sub-organs. Our findings show that a lack of Pax6 in these regions leads to dysgenesis of the iris and CB, while heterozygosity impedes growth of the iris and maturation of the iris sphincter. Overexpression of the canonical, but not the alternative splice variant of Pax6 results in severe structural aberrations of the CB and hyperplasia of the iris sphincter. A splice variant specific rescue experiment revealed that both splice variants are able to correct iris hypoplasia, while only the canonical form rescues the sphincter. Overall, these findings demonstrate the dosage-sensitive roles of Pax6 in the formation of both the CB and the iris. PMID- 19563799 TI - Pyruvate kinase isoenzyme M2 is a glycolytic sensor differentially regulating cell proliferation, cell size and apoptotic cell death dependent on glucose supply. AB - The glycolytic key regulator pyruvate kinase M2 (M2-PK or PKM2) can switch between a highly active tetrameric and an inactive dimeric form. The transition between the two conformations regulates the glycolytic flux in tumor cells. We developed specific M2-PK-binding peptide aptamers which inhibit M2-PK, but not the 96% homologous M1-PK isoenzyme. In this study we demonstrate that, at normal blood glucose concentrations, peptide aptamer-mediated inhibition of M2-PK induces a significant decrease of the population doubling (PDL rate) and cell proliferation rate as well as an increase in cell size, whereas under glucose restriction an increase in PDL and cell proliferation rates but a decrease in cell size was observed. Moreover, M2-PK inhibition rescues cells from glucose starvation-induced apoptotic cell death by increasing the metabolic activity. These findings suggest that M2-PK is a metabolic sensor which regulates cell proliferation, cell growth and apoptotic cell death in a glucose supply-dependent manner. PMID- 19563800 TI - Genomic response to Wnt signalling is highly context-dependent--evidence from DNA microarray and chromatin immunoprecipitation screens of Wnt/TCF targets. AB - Wnt proteins are important regulators of embryonic development, and dysregulated Wnt signalling is involved in the oncogenesis of several human cancers. Our knowledge of the downstream target genes is limited, however. We used a chromatin immunoprecipitation-based assay to isolate and characterize the actual gene segments through which Wnt-activatable transcription factors, TCFs, regulate transcription and an Affymetrix microarray analysis to study the global transcriptional response to the Wnt3a ligand. The anti-beta-catenin immunoprecipitation of DNA-protein complexes from mouse NIH3T3 fibroblasts expressing a fusion protein of beta-catenin and TCF7 resulted in the identification of 92 genes as putative TCF targets. GeneChip assays of gene expression performed on NIH3T3 cells and the rat pheochromocytoma cell line PC12 revealed 355 genes in NIH3T3 and 129 genes in the PC12 cells with marked changes in expression after Wnt3a stimulus. Only 2 Wnt-regulated genes were shared by both cell lines. Surprisingly, Disabled-2 was the only gene identified by the chromatin immunoprecipitation approach that displayed a marked change in expression in the GeneChip assay. Taken together, our approaches give an insight into the complex context-dependent nature of Wnt pathway transcriptional responses and identify Disabled-2 as a potential new direct target for Wnt signalling. PMID- 19563801 TI - Independent and cooperative action of Psen2 with Psen1 in zebrafish embryos. AB - Presenilin1 (PSEN1) and presenilin2 (PSEN2) are involved in the processing of type-1 transmembrane proteins including the amyloid precursor protein (APP), Notch and several others. PSEN1 has been shown to be crucial for proteolytic cleavage of Notch in developing animal embryos. Mouse embryos lacking Psen1 function show disturbed neurogenesis and somite formation, resembling Notch pathway mutants. However, loss of Psen2 activity reveals only a minor phenotype. Zebrafish embryos are a valuable tool for analysis of the molecular genetic control of cell differentiation since endogenous gene expression can be modulated in subtle and complex ways to give a phenotypic readout. Using injection of morpholino antisense oligonucleotides to inhibit protein translation in zebrafish embryos, we show that reduced Psen2 activity decreases the number of melanocytes in the trunk but not in the cranial area at 2 days post fertilisation (dpf). Reduced Psen2 activity apparently reduces Notch signalling resulting in perturbed spinal neurogenin1 (neurog1) expression, neurogenesis and trunk and tail neural crest development. Similar effects are seen for reduced Psen1 activity. These results suggest that Psen2 plays a more prominent role in Notch signalling and embryo development in zebrafish than in mammals. Intriguingly, decreased Psen2 activity increases the number of Dorsal Longitudinal Ascending (DoLA) interneurons in the spinal cord while decreased Psen1 activity has no effect. However, the effect on DoLAs of reduced Psen2 can be ameliorated by Psen1 loss. The effects of changes in Psen2 activity on DoLA interneurons and other cells in zebrafish embryos provide bioassays for more detailed dissection of Psen2 function. PMID- 19563802 TI - Oxytocin stimulates in vitro angiogenesis via a Pyk-2/Src-dependent mechanism. AB - We previously reported that the hypothalamic hormone oxytocin (OT), best known for its uterotonic activity, also stimulates migration and invasion in human umbilical vein endothelial cells (HUVECs), thus suggesting a possible role for the peptide in the regulation of angiogenesis. We identified the Gq coupling of OT receptors (OTRs) and phospholipase C (PLC) as the main effectors of OT's action in HUVECs. Moreover, the pro-migratory effect of OT required the OTR induced activation of the phosphatidylinositol-3-kinase (PI-3-K)/AKT/endothelial nitric oxide synthase (eNOS) pathway. To better characterize the proposed pro angiogenic effect of OT in HUVECs, we have now utilized a three-dimensional (3-D) in vitro angiogenesis assay, and demonstrated that OT stimulates the outgrowth of capillary-like structures from HUVEC spheroids to an extent comparable to that of vascular endothelial growth factor (VEGF). This OT effect was abolished by inhibitors of PLC, PI-3-K and Src kinase. It was also found that OT phosphorylates proline-rich tyrosine kinase-2 (Pyk-2) and Src kinase in a PLC- and calcium-dependent manner. Furthermore, knockdown of Pyk-2 expression by RNA interference markedly impaired Src phosphorylation, migration and endothelial cell sprouting induced by OT. In conclusion, by using a pharmacological and genetic approach, the OT pro-angiogenic action and the cascade of intracellular signals responsible for it were defined by showing for the first time that OT, by interacting with its Gq-coupled receptor, induces HUVEC capillary outgrowth via Pyk-2 phosphorylation, which activates Src which in turn activates the PI-3-K/AKT pathway. PMID- 19563803 TI - Neurobehavioral maturation of offspring from epileptic dams: study in the rat lithium-pilocarpine model. AB - In the present study, we explored the consequences of epilepsy on the neurobehavioral development of the offspring in a rat model of spontaneous epilepsy, the lithium-pilocarpine model of temporal lobe epilepsy not dependent on genetic factors and in animals not receiving any antiepileptic treatment. Status epilepticus was induced by lithium-pilocarpine in female rats. After the occurrence of spontaneous seizures the rats were mated and the neurobehavioral development of the offspring was explored. Rat pups were cross-fostered early after birth. We hence obtained pups born from or raised by epileptic or non epileptic dams. On the dams, we performed a follow-up of maternal care during pregnancy. On the pups, we performed a follow-up of classical parameters of development such as body weight and eyelid opening, and subjected the pups to various tests representative of neurobehavioral maturation extending from postnatal day 4 (PD4) to PD30 (righting reflex, suspension time, negative geotaxis, open field, locomotor coordination and eight arm maze). Altogether our data show that rat pups born from or raised by epileptic dams develop as well as control pups raised by control dams. Intriguingly, pups born from lithium pilocarpine exposed dams and raised by control mothers tend to have better scores than the two other groups in all tests. This indicates that the exposure to seizures during pregnancy is not harmful for the development of the fetus. PMID- 19563804 TI - Evaluation of Toxoplasma gondii placental transmission in BALB/c mice model. AB - Toxoplasma gondii infection is common worldwide and highly important to pregnant women as it can be transmitted to the fetus via the placenta. This study aimed at evaluating the prevention of placental transmission in two different strains after chronic infection with each one of the strains. A BALB/c mice model was inoculated 30days before breeding (immunization) and re-infected 12 and 15days after pregnancy (challenge). Seven experimental groups were assayed: G1: ME49 immunization (type II), M7741-challenge (type III); G2: M7741-immunization, ME49 challenge; G3, ME49-immunization; G4: M7741-immunization; G5: ME49-challenge; G6: M7741-challenge; G7: saline solution inoculation. Serology, mouse bioassay, PCR and RLFP of the uterus, placenta and fetus were performed to determine the congenital transmission of the strains challenged after chronic infection. IgG T. gondii antibodies were detected in G1, G2, G3 and G4, but not in G5, G6 and G7. All animals of G5 and G6 were IgM-positive. Congenital infection was not detected by bioassay and PCR. Nonetheless, placentas from G3 and G4 resulted positive but no corresponding fetal infection was detected. G1 and G2 did not show the genotype of the strain challenged during pregnancy, only those of chronic infection. Thus, the chronically infected BALB/c mice showed no re-infection after inoculation with another strain during pregnancy. Further studies with different parasite loads and different mice lineages are needed. PMID- 19563805 TI - Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection. AB - Specific primers to determine the presence of an intestinal fluke, Haplorchis taichui, were investigated using the high annealing temperature random amplified polymorphic DNA (HAT-RAPD) PCR, and 18 arbitrary primers (Operon Technologies), to generate different polymorphic DNA profiles. Thirteen kinds of parasites were used to compare fingerprints. A 256bp HAT-RAPD marker, generated from the OPP-11 primer, was found to be H. taichui-specific, and this marker was cloned, transformed, and sequenced. From the sequence data, a pair of primers were designed with Genetyx-MAC ver.11 and indicated as: Hap-t F 5'-GGC CAA CGC AAT CGT CAT CC-3' and Hap-t R 5'-GCG TCG GGT TTC AGA CAT GG-3'. These specific primers were tested for efficacy and specificity by amplifying them with all 13 parasites DNAs in PCR reaction. A 256bp amplicon was generated, which was shown to have a positive result, only for H. taichui DNA. It revealed no cross-reaction with any of the other tested parasite species. The minimum DNA template, needed for detection by PCR, was 0.1picogram (pg). The successful development of H. taichui specific primers is expected to be beneficial for epidemiological studies and for prevention and control of these parasitic infections. PMID- 19563806 TI - Thyroid hormones reverse the UV-induced repression of APP in neuroblastoma cells. AB - As a precursor of the neurotoxic amyloid-beta peptide, APP plays a central role in Alzheimer's disease. We have recently reported that the tumor suppressor p53 inhibits APP gene transcription through the same DNA sequences that mediate an inhibitory effect of thyroid hormones. Now, we have analyzed whether the thyroid hormone T3 can modulate the effects of p53 on APP expression. Exposition to UVC radiation leads to a marked decrease of intracellular APP levels that is paradoxically reversed by T3. Repression by UVC and reversion by the hormone are not observed in cells depleted of p53, demonstrating a p53-dependent mechanism. These results suggest the existence of a cross-talk between p53 and T3 that could play an important role in Alzheimer s disease. PMID- 19563807 TI - N-terminal amphipathic helix as a trigger of hemolytic activity in antimicrobial peptides: a case study in latarcins. AB - In silico structural analyses of sets of alpha-helical antimicrobial peptides (AMPs) are performed. Differences between hemolytic and non-hemolytic AMPs are revealed in organization of their N-terminal region. A parameter related to hydrophobicity of the N-terminal part is proposed as a measure of the peptide propensity to exhibit hemolytic and other unwanted cytotoxic activities. Based on the information acquired, a rational approach for selective removal of these properties in AMPs is suggested. A proof of concept is gained through engineering specific mutations that resulted in elimination of the hemolytic activity of AMPs (latarcins) while leaving the beneficial antimicrobial effect intact. PMID- 19563808 TI - FACS-assisted microarray profiling implicates novel genes and pathways in zebrafish gastrointestinal tract development. AB - BACKGROUND & AIMS: The zebrafish Danio rerio is an excellent model system for mammalian gastrointestinal development. To identify differentially regulated genes important in gastrointestinal organogenesis, we profiled the transcriptome of the zebrafish developing gastrointestinal tract. METHODS: Embryos from a transgenic zebrafish line expressing green fluorescent protein (GFP) in the developing intestine, liver, and pancreas were dissociated at 4 developmental time points, their cells sorted based on GFP expression with fluorescence activated cell sorting (FACS), and analyzed with microarrays. To improve our analysis, we annotated the Affymetrix Zebrafish GeneChip with human orthologs. RESULTS: Transcriptional profiling showed significant differences between GFP(+) and GFP(-) cells. Up-regulated genes and pathways were consistent with mammalian gastrointestinal development, such as hepatic nuclear factor gene networks and cancer. We implicate the phosphatidylinositol 3 kinase (PI3K) pathway and show that inhibition with LY294002 causes gastrointestinal defects in zebrafish. We identified novel genes, such as the microRNAs miR-217 and miR-122, the tight junction protein claudin c, the gene fam136a, and a zebrafish tetraspanin. Novel pathways include genes containing a putative transcription factor binding sequence, GGAANCGGAANY, and a nucleolar gene network. The zebrafish microarrays also identify a set of 32 genes that may mediate the effects of gain of chromosome arm 8q in human colon, liver, and pancreatic cancers. CONCLUSIONS: We successfully combine FACS and microarray profiling to follow organogenesis throughout development. These experiments identify novel genes and pathways that probably play a role in mammalian gastrointestinal development and are potential targets for therapeutic intervention in the management of gastrointestinal disease and cancer. PMID- 19563810 TI - Non-receptor tyrosine kinases c-Abl and Arg regulate the activity of C/EBPbeta. AB - The CCAAT/enhancer-binding protein beta (C/EBPbeta) is a critical transcription factor that regulates gene expression during numerous biological processes, including differentiation, metabolism, homeostasis, proliferation, tumorigenesis, inflammation, and apoptosis. In this study, interactions between C/EBPbeta and either the Abelson murine leukemia viral oncogene homolog 1 (c-Abl) or the Abl related gene (Arg) were demonstrated in vitro and in vivo with a direct binding assay and by co-immunoprecipitation, respectively. The Y79 amino acid residue of C/EBPbeta was phosphorylated by c-Abl or Arg. The phosphorylation of C/EBPbeta resulted in an increased C/EBPbeta stability and a potentiation of C/EBPbeta transcription activation activity in cells. Expression of the C/EBPbeta(Y79F) mutant in HEK293, and K562, and in other cell lines, resulted in less of a delay in the cell cycle compared to the wild type C/EBPbeta; furthermore, the C/EBPbeta (Y79F) mutant induced an increased apoptosis compared to the wild type C/EBPbeta. These findings suggest that the c-Abl family non-receptor tyrosine kinases have a role in the regulation of the C/EBPbeta transcription factor. PMID- 19563811 TI - Structure-function studies of a Melanocarpus albomyces laccase suggest a pathway for oxidation of phenolic compounds. AB - Melanocarpus albomyces laccase crystals were soaked with 2,6-dimethoxyphenol, a common laccase substrate. Three complex structures from different soaking times were solved. Crystal structures revealed the binding of the original substrate and adducts formed by enzymatic oxidation of the substrate. The dimeric oxidation products were identified by mass spectrometry. In the crystals, a 2,6-dimethoxy-p benzoquinone and a C-O dimer were observed, whereas a C-C dimer was the main product identified by mass spectrometry. Crystal structures demonstrated that the substrate and/or its oxidation products were bound in the pocket formed by residues Ala191, Pro192, Glu235, Leu363, Phe371, Trp373, Phe427, Leu429, Trp507 and His508. Substrate and adducts were hydrogen-bonded to His508, one of the ligands of type 1 copper. Therefore, this surface-exposed histidine most likely has a role in electron transfer by laccases. Based on our mutagenesis studies, the carboxylic acid residue Glu235 at the bottom of the binding site pocket is also crucial in the oxidation of phenolics. Glu235 may be responsible for the abstraction of a proton from the OH group of the substrate and His508 may extract an electron. In addition, crystal structures revealed a secondary binding site formed through weak dimerization in M. albomyces laccase molecules. This binding site most likely exists only in crystals, when the Phe427 residues are packed against each other. PMID- 19563809 TI - Structure and uncoating of immature adenovirus. AB - Maturation via proteolytic processing is a common trait in the viral world and is often accompanied by large conformational changes and rearrangements in the capsid. The adenovirus protease has been shown to play a dual role in the viral infectious cycle: (a) in maturation, as viral assembly starts with precursors to several of the structural proteins but ends with proteolytically processed versions in the mature virion, and (b) in entry, because protease-impaired viruses have difficulties in endosome escape and uncoating. Indeed, viruses that have not undergone proteolytic processing are not infectious. We studied the three-dimensional structure of immature adenovirus particles as represented by the adenovirus type 2 thermosensitive mutant ts1 grown under non-permissive conditions and compared it with the mature capsid. Our three-dimensional electron microscopy maps at subnanometer resolution indicate that adenovirus maturation does not involve large-scale conformational changes in the capsid. Difference maps reveal the locations of unprocessed peptides pIIIa and pVI and help define their role in capsid assembly and maturation. An intriguing difference appears in the core, indicating a more compact organization and increased stability of the immature cores. We have further investigated these properties by in vitro disassembly assays. Fluorescence and electron microscopy experiments reveal differences in the stability and uncoating of immature viruses, both at the capsid and core levels, as well as disassembly intermediates not previously imaged. PMID- 19563812 TI - The genesis of ribosome structure: how a protein generates RNA structure in real time. AB - Ribosomal subunit assembly is initiated by the binding of several primary binding proteins. Results from chemical modification studies show that 16S ribosomal RNA undergoes striking structural rearrangements when protein S17 is bound. For the first time, we are able to distinguish and order these structural rearrangements by using time-dependent chemical probing. Initially, protein S17 binds to a portion of helix 11, inducing a kink-turn in that helix that bends helix 7 toward the S17-helix 11 complex in a hairpin-like manner, allowing helix 7 to bind to protein S17. This structural change is rapidly stabilized by interactions at the distal and proximal ends of both RNA helices. Identifying the dynamic nature of interactions between RNA and proteins is not only essential in unraveling ribosome assembly, but also has more general application to all protein-RNA interactions. PMID- 19563814 TI - Advantage of having regulatory T cells requires localized suppression of immune reactions. AB - The immune system of vertebrates may attack its own body and cause autoimmunity diseases. To prevent autoimmunity, regulatory T cells suppress the activity of the autoreactive effector T cells, but they also interrupt normal immune reactions against foreign antigens. In this paper, we discuss the advantage of having some regulatory T cells by considering the host's ability of coping with foreign antigens and the harm of autoimmunity. Assumptions are as follows: the immature T cells reactive to abundant self-antigens are eliminated, those reactive to rare self-antigen will become regulatory T cells, and those that fail to interact with the antigens to which they are reactive will become effector T cells. Some self-reactive immature T cells may fail to interact with their own target antigens during the limited training period, and will later become effector T cells, causing autoimmunity. Analysis suggests that, having some regulatory T cells can never be advantageous to the host, if activated regulatory T cells suppress effector T cells at any location of the body (global suppression). In contrast, producing some regulatory T cells can be beneficial, if the body is composed of many compartments and regulatory T cells suppress the immune reactions only within the same compartment (localized suppression). This requires regulatory T cells to stop circulating once they are activated by their own target self-antigens. PMID- 19563813 TI - Remote homology between Munc13 MUN domain and vesicle tethering complexes. AB - Most core components of the neurotransmitter release machinery have homologues in other types of intracellular membrane traffic, likely underlying a universal mechanism of intracellular membrane fusion. However, no clear similarity between Munc13s and protein families generally involved in membrane traffic has been reported, despite the essential nature of Munc13s for neurotransmitter release. This crucial function was ascribed to a minimal Munc13 region called the MUN domain, which likely participates in soluble N-ethylmaleimide sensitive factor attachment protein receptor complex (SNARE) assembly and is also found in Ca(2+) dependent activator protein for secretion. We have now used comparative sequence and structural analyses to study the structure and evolutionary origin of the MUN domain. We found weak yet significant sequence similarities between the MUN domain and a set of protein subunits from several related vesicle tethering complexes, such as Sec6 from the exocyst complex and Vps53 from the Golgi associated retrograde protein complex. Such an evolutionary relationship allows structure prediction of the MUN domain and suggests functional similarities between MUN domain-containing proteins and multisubunit tethering complexes such as exocyst, conserved oligomeric Golgi complex, Golgi-associated retrograde protein complex, and Dsl1p. These findings further unify the mechanism of neurotransmitter release with those of other types of intracellular membrane traffic and, in turn, support a role for tethering complexes in soluble N ethylmaleimide sensitive factor attachment protein receptor complex assembly. PMID- 19563815 TI - The roles of predator maturation delay and functional response in determining the periodicity of predator-prey cycles. AB - Population cycles in small mammals have attracted the attention of several generations of theoretical and experimental biologists and continue to generate controversy. Top-down and bottom-up trophic regulations are two recent competing hypotheses. The principal purpose of this paper is to explore the relative contributions of a variety of ecological factors to predator-prey population cycles. Here we suggest that for some species - collared lemmings, snowshoe hares and moose in particular - maturation delay of predators and the functional response of predation appear to be the primary determinants. Our study suggests that maturation delay alone almost completely determines the cycle period, whereas the functional response greatly affects its amplitude and even its existence. These results are obtained from sensitivity analysis of all parameters in a mathematical model of the lemming-stoat delayed system, which is an extension of Gilg's model. Our result may also explain why lemmings have a 4-year cycle whereas snowshoe hares have a 10-year cycle. Our parameterized model supports and extends May's assertion that time delay impacts cycle period and amplitude. Furthermore, if maturation periods of predators are too short or too long, or the functional response resembles Holling Type I, then population cycles do not appear; however, suitable intermediate predator maturation periods and suitable functional responses can generate population cycles for both prey and predators. These results seem to explain why some populations are cyclic whereas others are not. Finally, we find parameterizations of our model that generate a 38-year population cycle consistent with the putative cycles of the moose-wolf interactions on Isle Royale, Michigan. PMID- 19563816 TI - Role of PPAR-gamma in inflammation. Prospects for therapeutic intervention by food components. AB - Peroxisome proliferator activated receptor gamma (PPARgamma) is a ligand dependent transcription factor and a member of the nuclear receptor superfamily. Acting as sensors of hormones, vitamins, endogenous metabolites and xenobiotic compounds, the nuclear receptors control the expression of a very large number of genes. PPARgamma has been known for some time to regulate adipocyte differentiation, fatty acid storage and glucose metabolism, and is a target of anti-diabetic drugs. More recently, PPARgamma has been recognized as playing a fundamentally important role in the immune response through its ability to inhibit the expression of inflammatory cytokines and to direct the differentiation of immune cells towards anti-inflammatory phenotypes. A feature of PPARgamma is the structural diversity of its ligands, which encompass endogenous metabolites, dietary compounds and synthetic drugs. The high and increasing incidence of inflammatory and allergic disease, coupled with encouraging results from recent clinical trials, suggest that natural PPARgamma agonists found in foods may be beneficial to human health by acting as anti inflammatory molecules. PPARgamma is therefore not only a target of the pharmaceutical industry, but also of great potential interest to the food industry, since it is activated by several natural dietary constituents. The prospects for dietary intervention in inflammatory disease have improved somewhat over the last few years, and are reviewed here. PMID- 19563817 TI - Optic ataxia and the function of the dorsal stream: contributions to perception and action. AB - Optic ataxia (OA) is one of the symptoms pertaining to Balint's Syndrome. It has been described clinically for nearly 80 years before it became a cornerstone of the most popular dual stream theory of the visual brain. Over the last 10 years a regain of interest for this neurological condition lead to a number of precise analyses of the deficits found in optic ataxia, giving rise to a renewed outline of its very definition and hence of the function(s) of the occipito-parietal (dorsal) stream. In absence of concomitant clinical symptoms, we review evidence that misreaching errors in central vision result from the "hand effect": an erroneous dynamic spatial processing of proprioceptive information from the hand. When visual feedback of the hand is provided (closed-loop condition), pure optic ataxia is restricted to peripheral vision. This central versus peripheral vision distinction is repeatedly used to argue that action and perception are not unique and dissociated systems. New assessments of optic ataxia patients are provided, confirming on one hand that their visuomotor deficit is specific to peripheral vision (i.e. when the gaze and the hand goals are dissociated), on the other hand that they disclose perceptual deficits in peripheral vision. These results are coherent with the recent demonstration that optic ataxia patients exhibit a general contralesional deficit for dynamic visuo-spatial processing, affecting both hand and eye movements [Gaveau, V., Pelisson, D., Blangero, A., Urquizar, C., Prablanc, C.,Vighetto, A., et al. (2008). A common parietal module for saccade and reach: Eye-hand coordination and saccadic control in optic ataxia. Neuropsychologia, 46, 475-486]. Such module(s) within the dorsal stream could be used for both action and perception in the periphery. It is concluded that optic ataxia cannot be considered as a unitary and specific visuo-manual deficit, and that the modular organisation of the dorsal stream allows for numerous dorsal ventral interactions for perception and action. PMID- 19563818 TI - Opposite hemispheric asymmetries for pitch identification in absolute pitch and non-absolute pitch musicians. AB - The aim of the present study is to investigate functional laterality for pitch identification in subjects with absolute pitch (AP) and without absolute pitch (NAP). Forty-four musicians were divided into two groups (AP and NAP) on the basis of their performance in a preliminary standard AP-test. They were subsequently presented with an AP-test designed for dichotic listening, a neuropsychological technique which allows the investigation of functional hemispheric asymmetries. Dependent variables were accuracy and reaction time. It was observed that AP and NAP subjects exhibit opposite hemispheric specialization for pitch identification, AP subjects showing a bias towards the left hemisphere and NAP subjects showing a bias towards the right hemisphere. Results are discussed in the context of neuroimaging evidence of structural and functional asymmetries related to AP. PMID- 19563819 TI - Object priming and recognition memory: dissociable effects in left frontal cortex at encoding. AB - Functional magnetic resonance imaging (fMRI) studies have implicated the left prefrontal cortex in priming. We tested the hypothesis that object encoding activity in different prefrontal cortex regions selectively predicts subsequent object priming and recognition respectively. Participants were scanned whilst making semantic category judgements about novel object pictures. One week later priming and recognition of these objects were tested. Encoding that produced long lasting priming in the absence of recognition memory was associated with increased activity in left inferior prefrontal (BA 47) and superior frontal (BA 8) cortices. In contrast, encoding that produced object recognition one week later activated the left middle frontal cortex (BA 9). This is consistent with other evidence indicating that object priming and recognition are independent kind of memory. Problems of measuring item-by-item recognition and priming together are discussed. PMID- 19563820 TI - Purification, cloning and characterization of fragaceatoxin C, a novel actinoporin from the sea anemone Actinia fragacea. AB - Actinia fragacea is commonly called the "strawberry" anemone because of the distinctive yellow or green spots displayed on its red column. Its venom contains several haemolytic proteins with a molecular mass of approximately 20 kDa that can be separated by ion-exchange column chromatography. One of them was purified to homogeneity and was named fragaceatoxin C (FraC). Its 15 N-terminal residues were identified by Edman degradation and served to obtain its complete DNA coding sequence by RT-PCR. The coding region of FraC was amplified and cloned in the expression vector pBAT-4. Purified recombinant FraC consists of 179 amino acids and multiple sequence alignment with other actinoporins clearly indicates that FraC belongs to this protein family. The secondary structure, thermal stability and lytic activity of native and recombinant FraC were practically identical and exhibit the same basic features already described for equinatoxin-II and sticholysin-II. PMID- 19563821 TI - Study of patient's injuries by stingrays, lethal activity determination and cardiac effects induced by Himantura gerrardi venom. AB - Stingrays are common inhabitants in the northern waters of the Persian Gulf and Oman Sea. In the present study, the clinical aspects of injuries induced in three patients bitten by stingrays in Hormozgan province waters were first examined. The LD(50) of crude venom extract obtained from the most common stingray in Hormozgan province (Himantura gerrardi) was then estimated by up-and-down dosing and double dose methods in mice. Third and finally, the cardiac symptoms induced by injection of the extract from the venomous spines were evaluated in rats. Intense pain was noticed in all human cases. Redness was observed in two cases, and spasm and seizure were each recorded in only one case. LD(50) of the venom extract in mice was about 100 mg kg(-1). The observed cardiac symptoms in rats included an increase in pulse rate and various changes in electrocardiogram (ECG) parameters such as T and Q values, and PR and RR intervals. PMID- 19563822 TI - Natural history of the terciopelo Bothrops asper (Serpentes: Viperidae) in Costa Rica. AB - The terciopelo Bothrops asper is the only lancehead species widely distributed in the humid lowlands of Middle America and northwestern South America. Its large body size, relative abundance and cryptic habits contribute to the high incidence of snakebites induced by this species throughout its distribution. The terciopelo plays an important role in ecosystems, both as prey and as a generalist predator. Diet comprises a great variety of prey items, including some species that are considered nuisances. B. asper, as other lancehead species, exhibits a notable ontogenetic shift in diet, consuming ectotherms (mainly frogs and lizards) when young, and increasingly incorporating birds, rodents, and other small mammals with maturity. Adult terciopelos also consume large anurans, especially when endothermic prey availability is low. Using radiotelemetry we determined home range and movement patterns from 28 individual B. asper at La Selva Biological Station, Costa Rica. Overall home range estimates are relatively small compared with other pitvipers, averaging between 3.71ha and 5.95ha; home range size did not differ between males and females. Movement patterns are largely aseasonal and consist of short (<10 m) movements between daytime shelter and nocturnal ambush sites within a given area, interspersed with longer distance (>50 m) movements to new foraging areas. Habitat use is related to prey availability and therefore to foraging strategy. Our data support a strong preference for areas near swamps by both sexes. Reproduction in B. asper is highly seasonal, and--apparently- biannual. Reproductive cycles in Costa Rica are tightly related to rainfall patterns. Therefore, the timing of breeding differs between populations in the Caribbean and Pacific lowlands. Bothrops asper is adapted to areas with low levels of disturbance along the agricultural frontier, and consequently it is not rare to find it in or near human dwellings. However, despite popular belief, no evidence supports a purported increase in population density of this species in Costa Rica. Despite human persecution and substantial modification of habitat, B. asper is a species with a conservation status of least concern, and probably will likely persist well into the future. Thus, it is important to learn how to coexist with this species, and to improve mechanisms for the prevention and treatment of accidental snakebite and its consequences. PMID- 19563823 TI - Time-dependent differential gene expression in lysophosphatidic acid-treated young and senescent human diploid fibroblasts. AB - The gene expression profiles of lysophosphatidic acid (LPA)-treated young and senescent human diploid fibroblasts (HDFs) were examined using cDNA microarray analysis. The expression of some genes, including EGR 1/3 and MRRF, was controlled by LPA similarly in young and senescent cells, showing a typical time dependent up-and-down expression profile. In contrast, some other genes, including DUSP6, CYR61, and F3, showed sustained upregulation in senescent HDFs later after LPA treatment. These genes might be involved in altered LPA responsiveness during the aging process. PMID- 19563825 TI - Genetic association studies of performance monitoring and learning from feedback: the role of dopamine and serotonin. AB - Performance monitoring is essential for optimization of action outcomes. Research consistently implicates the posterior medial frontal cortex, particularly the rostral cingulate zone, in monitoring for unfavorable action outcomes, signaling the need for adjustments and learning from feedback. Current theories suggest that phasic dopaminergic signals coding unexpected positive or negative outcomes play a major role in this function. Here, I review EEG, neuroimaging and computational modeling studies making use of polymorphisms of candidate genes affecting neurotransmission, with a specific focus on dopamine. Although the evidence is still rather sparse, findings speak for a prominent role of dopamine in performance monitoring. However, the exact function in cortical areas underlying this function, such as the rostral cingulate zone, remains to be determined. Different hypotheses on the interaction of the rostral cingulate zone, the striatum, and the mesencephalic dopaminergic nuclei are discussed. PMID- 19563824 TI - Membrane translocation by anthrax toxin. AB - Much attention has been focused on anthrax toxin recently, both because of its central role in the pathogenesis of Bacillus anthracis and because it has proven to be one of the most tractable toxins for studying how enzymic moieties of intracellularly acting toxins traverse membranes. The Protective Antigen (PA) moiety of the toxin, after being proteolytically activated at the cell surface, self-associates to form a heptameric pore precursor (prepore). The prepore binds up to three molecules of Edema Factor (EF), Lethal Factor (LF), or both, forming a series of complexes that are then endocytosed. Under the influence of acidic pH within the endosome, the prepore undergoes a conformational transition to a mushroom-shaped pore, with a globular cap and 100A-long stem that spans the membrane. Electrophysiological studies in planar bilayers indicate that EF and LF translocate through the pore in unfolded form and in the N- to C-terminal direction. The pore serves as an active transporter, which translocates its proteinaceous cargo across the endosomal membrane in response to DeltapH and perhaps, to a degree, Deltapsi. A ring of seven Phe residues (Phe427) in the lumen of the pore forms a seal around the translocating polypeptide and blocks the passage of ions, presumably preserving the pH gradient. A charge state dependent Brownian ratchet mechanism has been proposed to explain how the pore translocates EF and LF. This transport mechanism of the pore may function in concert with molecular chaperonins to effect delivery of effector proteins in catalytically active form to the cytosolic compartment of host cells. PMID- 19563827 TI - Pharmacogenomics of antidepressant drugs. AB - While antidepressant pharmacotherapy is an effective treatment of depression, it is still hampered by the slow onset of appreciable clinical improvement and a series of side effects. Moreover, a substantial group of patients does not achieve remission or fails to respond at all. One possible source accounting for these variations in treatment outcome are genetic differences. In recent years a number of pharmacogenetic studies on antidepressant drugs have been published. This manuscript summarizes findings related to the pharmacogenetics of genes involved in the pharmacokinetics as well as pharmacodynamics of antidepressants to date. Illustrated by examples from current candidate gene- and whole genome association studies, this manuscript critically discusses aspects of pharmacogenetic studies in antidepressant response related to study design and clinical relevance. PMID- 19563826 TI - Therapeutic potential of dithiolethiones for hepatic diseases. AB - Comprehensive studies support the notion that oltipraz [4-methyl-5-(2-pyrazynyl) 1,2-dithiole-3-thione] and its congeners exert cancer chemopreventive effects by the prevention, inhibition or reversal of carcinogenic processes. Recently, it was found that dithiolethione compounds had the activities to prevent or treat fibrosis, insulin resistance, and mitochondrial protective effects in the liver by a mechanism involving AMP-activated protein kinase (AMPK) and/or 70-kDa ribosomal protein S6 kinase 1 (S6K1). Moreover, chemical regulation of the AMPK S6K1 pathway was found to affect Liver X receptor (LXR) activity and lipogenesis, leading to the identification of AMPK and S6K1 as targets for treating hepatic steatosis. These biological activities of dithiolethiones may offer a novel approach to pharmaceutical intervention. This review focuses on the interaction between oltipraz and the AMPK-mTOR-S6K1 pathway, which regulates genes that confer hepatocyte protection from intoxication, disrupted energy metabolism, and inflammation. In terms of therapeutic potential, the findings reviewed here demonstrate a new therapeutic potential for dithiolethiones, which function in a unique manner, and offer the possibility of new treatments for hepatic diseases. PMID- 19563828 TI - Multiplex real-time RT-PCR for simultaneous detection of GI/GII noroviruses and murine norovirus 1. AB - A quantitative two-step multiplex real-time reverse transcriptase (RT-) PCR assay for the simultaneous detection of genogroup I (GI) and genogroup II (GII) noroviruses (NoVs) is described below. A murine norovirus 1 (MNV-1) real-time PCR detection assay described recently was integrated successfully into the multiplex assay, making it possible to detect GI and GII NoVs and MNV-1 in one reaction tube with MNV-1 plasmid DNA as real-time PCR internal amplification control (IAC). The results showed a nearly complete concordance between the multiplex assay and the corresponding single-target PCRs. Analysis of competition between the individual reactions within the multiplex real-time PCR assay showed that GI and GII NoV plasmid DNAs mixed at equimolar concentrations were detected reproducibly and quantitatively, while a 4 log excess between GI and GII plasmid DNAs hindered amplification of the target with the lowest concentration. High concentrations of the real-time PCR IAC (MNV-1 plasmid DNA) also interfered with the possibility of the developed multiplex real-time RT-PCR assay to detect quantitatively and simultaneously the presence of GI and GII NoVs within one sample. The specificity of the multiplex assay was evaluated by testing a NoV RNA reference panel containing nine GI, eight GII, and one GIV in vitro synthesized RNA fragment, plus 16 clinical samples found positive for GI and GII NoVs previously. In addition, a collection of bovine NoVs and other (non-NoV) enteric viruses were found to be negative, and no cross-amplification between genogroups was observed. PMID- 19563830 TI - Molecular basis of binding of the Plasmodium falciparum receptor BAEBL to erythrocyte receptor glycophorin C. AB - Plasmodium falciparum invades human erythrocytes by redundant pathways. Unlike Plasmodium vivax that has one Duffy Binding-Like (DBL) receptor, P. falciparum has four members of the DBL receptor family. Furthermore, one of these DBL genes, BAEBL, has polymorphisms at four amino acids in region II; each polymorphism binds to a different erythrocyte receptor. One BAEBL variant (VSTK) binds specifically to erythrocyte glycophorin C and binds poorly to neuraminidase treated erythrocytes. When the amino acid threonine (T121) in BAEBL (VSTK) is changed to a lysine (VSKK), it no longer requires sialic acid as a receptor. To explore the molecular basis of sialic acid binding, we modeled the structure of region II of BAEBL (VSTK) on the crystal structure of a related DBL receptor, region II of erythrocyte binding antigen-175 (EBA-175). Four charged amino acids, R52, R114, E54 and D125, are predicted to surround T121 in BAEBL (VSTK). They were individually mutated to alanine (R52A, R114A, E54A, and D125A) or lysine (R52K, R114K) and expressed on the surface of Chinese hamster ovary (CHO-K1) cells. BAEBL (VSTK) with mutations in R52 or R114 of BAEBL (VSTK) bound neuraminidase-treated erythrocytes. Unlike the arginine mutations, E54A and D125A still bound poorly to neuraminidase-treated erythrocytes. These findings suggest that the two arginine residues surrounding T121 are critical for the binding specificity of BAEBL (VSTK) to sialic acid and suggest a role for arginine in sialic acid binding independent of its negative charge. PMID- 19563831 TI - Flow cytometric quantification of chlamydial infection in cell culture. AB - A flow cytometric method was developed, which allows fast and efficient analysis of cell cultures infected with chlamydiae. The proportion of positive cells increased with the infectious dose and correlated with chlamydia copy numbers calculated from real-time PCR. While retaining the advantages of single-cell analysis, flow cytometry allows handling of large sample numbers and counterstaining for additional marker proteins. PMID- 19563829 TI - Vaccinia virus vaccines: past, present and future. AB - Vaccinia virus (VACV) has been used more extensively for human immunization than any other vaccine. For almost two centuries, VACV was employed to provide cross protection against variola virus, the causative agent of smallpox, until the disease was eradicated in the late 1970s. Since that time, continued research on VACV has produced a number of modified vaccines with improved safety profiles. Attenuation has been achieved through several strategies, including sequential passage in an alternative host, deletion of specific genes or genetic engineering of viral genes encoding immunomodulatory proteins. Some highly attenuated third- and fourth-generation VACV vaccines are now being considered for stockpiling against a possible re-introduction of smallpox through bioterrorism. Researchers have also taken advantage of the ability of the VACV genome to accommodate additional genetic material to produce novel vaccines against a wide variety of infectious agents, including a recombinant VACV encoding the rabies virus glycoprotein that is administered orally to wild animals. This review provides an in-depth examination of these successive generations of VACV vaccines, focusing on how the understanding of poxviral replication and viral gene function permits the deliberate modification of VACV immunogenicity and virulence. PMID- 19563833 TI - Statins and cancer risk: a confounded association. PMID- 19563834 TI - Infection, inflammation, and homeostasis in inflammatory bowel disease. PMID- 19563835 TI - Hepatitis B virus immunotolerant patients: need to differentiate patients with or without liver disease. PMID- 19563836 TI - Endoscopic oncology: bridging the gap between gastroenterology and oncology. PMID- 19563837 TI - Gumming up the works: DNA polymers as HCV entry inhibitors. PMID- 19563838 TI - Do diabetes drugs modify the risk of pancreatic cancer? PMID- 19563841 TI - Gut permeability in irritable bowel syndrome: more leaks add to slightly inflamed bowel syndrome conspiracy theory. PMID- 19563842 TI - Targeting farnesoid x receptor in hepatic and biliary inflammatory diseases. PMID- 19563840 TI - Microbiome analysis in the esophagus. PMID- 19563843 TI - Occludin a new HCV entry factor: is the list of essential hepatitis C virus receptors complete? PMID- 19563844 TI - Chemoprevention in the lynch syndrome: what can we do? PMID- 19563845 TI - New insights into the role of Hedgehog signaling in gastrointestinal development and cancer. PMID- 19563846 TI - "Green odor" inhalation by rats down-regulates stress-induced increases in Fos expression in stress-related forebrain regions. AB - In the present study, on rats, a quantitative analysis of Fos protein immunohistochemistry was performed as a way of investigating the effects of inhalation of green odor (a mixture of equal amounts of trans-2-hexenal and cis-3 hexenol) on the neuronal activations in stress-related forebrain regions induced by acute and repeated stress. Rats were exposed to restraint stress for 90 min each day for 1, 2, 4, 7, or 11 consecutive days. The hypothalamic paraventricular nucleus (PVN), amygdala, hippocampus and paraventricular thalamic nucleus (PVT) were examined. Both acute and repeated restraint stress increased Fos-positive cells in the entire hypothalamic PVN, in the central and medial amygdala, and in PVT, although these responses declined upon repeated exposure to such stress. The stress-induced Fos responses were much weaker in rats that inhaled green odor during each day's restraint. No increases in Fos-positive cells were observed in the hippocampus in acutely stressed rats. The Fos-immunoreactive response to acute stress shown by the piriform cortex did not differ significantly between the vehicle+stress and green+stress groups. Green odor had inhibitory effects on the stress-induced corticosterone response, body-weight loss, and adrenal hypertrophy. These results suggest that in rats, green odor inhalation may, in an as yet unknown way, act on the brain to suppress activity in the neuronal networks involved in stress-related responses (such as activation of the hypothalamo-pituitary-adrenocortical axis and activation of the sympathetic nervous system, as well as stress-induced fear responses). PMID- 19563847 TI - Engineering Escherichia coli for an efficient aerobic fermentation platform. AB - Acetate, as a major by-product, was excreted by Escherichia coli when aerobic fermentation runs at high growth rates. In order to reduce the acetate secretion during the fermentation fundamentally, a list of genes related to acetate accumulation in E. coli was selected and knocked out. Physiological characterization of each mutant demonstrated that the growth and metabolites accumulation properties of these mutations exhibited significant change upon pathway engineering. The final engineered E. coli QZ1110 with ptsG, poxB, pta and iclR gene mutations was confirmed to accumulate 270% more biomass with 90% less acetate secretion than that of wild type E. coli in LB medium supplied with 1% glucose. Polyhydroxybutyrate biosynthesis experiment showed that the acetate reduction of the engineered strain in minimal medium also reduced 90% while the PHB accumulation increased almost 100% compare to wild type E. coli. PMID- 19563848 TI - Ischemic heart diseases: current treatments and future. AB - Ischemic heart disease is a rapidly increasing common cause of death in the world. This disease is the insufficient status of oxygen within the cardiac muscles due to an imbalance between oxygen supply and demand, and a cardiac disease that occurs as a result of coronary artery stenosis. Conventional surgery based therapy for the treatment of ischemic heart diseases has been advanced with biopharmaceutical-based therapy, such as protein, gene and cell therapy. The conventional medical therapy focuses on the use of drug eluting stents, coronary artery bypass-graft surgery and anti-thrombosis. Biopharmaceutical-based therapies including recombinant protein therapy, gene therapy and cell transplantation are recognized as promising approaches in inducing neovascularization and improving collateral blood flow in the ischemic heart. This review explores the current status and future of the treatment of ischemic heart diseases with conventional medical therapy, biopharmaceutical-based therapy focused on the proteins and polymeric hydrogels for delivery of therapeutic proteins. PMID- 19563849 TI - Enterostatin alters protein trafficking to inhibit insulin secretion in Beta-TC6 cells. AB - Enterostatin is a peptide that regulates dietary fat intake in rodents and inhibits insulin secretion from pancreatic beta cells. Microarray studies of the genomic response of both a human hepatoma cell line (HepG2 cells) and a mouse hypothalamic cell line (GT1-7 cells) to enterostatin suggested that it might regulate protein trafficking. Using semi-quantitative real-time PCR and Western blot analysis, we confirmed that enterostatin upregulated Scamp2 and down regulated Dynamin2 in these cell lines. The receptor for enterostatin is the F1 ATPase beta subunit. We transfected HepG2 cells with either a green fluorescent protein (GFP) tagged F1-ATPase beta subunit or a red fluorescent protein (RFP) tagged F1-ATPase alpha subunit to study the effects of enterostatin on translocation of its own receptor protein. Enterostatin induced movement of GFP beta subunit to the cell periphery area but did not have any effect on the localization of RFP-alpha subunit protein in HepG2. As Scamp2 is involved in glucose uptake in mouse Beta-TC6 insulinoma cells we tested enterostatin's effect in Beta-TC6 cells. Glucose stimulated insulin release was inhibited by enterostatin as reported previously. Using siRNA to Scamp2 did not change glucose stimulated insulin release but siRNA to Dynamin2 and dominant negative Dynamin2 (Dyn K44A) inhibited glucose stimulated insulin release and abolished the response to enterostatin. This suggests enterostatin inhibits glucose stimulated insulin release in pancreatic beta cells through down regulation of Dynamin2. This study also suggests that enterostatin might have a more generalized effect on protein trafficking in various cells. PMID- 19563851 TI - Evidence for neuroprotection by the fenamate NSAID, mefenamic acid. AB - Fenamate NSAIDs are inhibitors of cyclooxygenases, antagonists of non-selective cation channels, subtype-selective modulators of GABA(A) receptors, weak inhibitors of glutamate receptors and activators of some potassium channels. These pharmacological actions are all implicated in the pathogenesis of ischemic stroke. The aim of this study was to investigate the hypothesis that the fenamate, mefenamic acid, is neuroprotective in an in vitro and in vivo model of stroke. Embryonic rat hippocampal neurons were cultured and maintained for up to 14 days in vitro. At 9 or 14 days, cells were exposed to glutamate (5microM) or glutamate (5microM) plus mefenamic acid (10-100microM) or the control agent, MK 801 (10microM) for 10min. 24h later, cell death was determined by measuring lactate dehydrogenase (LDH) levels in the culture media. In vivo, male Wistar rats (300-350g) were subjected to 2h middle cerebral artery occlusion (MCAO) followed by 24h reperfusion. Animals received either a single i.v. dose of MFA (10mg/kg or 30mg/kg), or MK-801 (2mg/kg) or saline prior to MCAO or, four equal doses of MFA (20mg/kg) at 1h intervals beginning 1h prior to MCAO. Ischemic damage was then assessed 24h after MCAO. In vitro, mefenamic acid (10-100microM) and MK-801 (10microM) significantly reduced glutamate-evoked cell death compared with control cultures. In vivo, MFA (20mg/kgx4) significantly reduced infarct volume, total ischemic brain damage and edema by 53% (p< or =0.02), 41% (p< or =0.002) and 45% (p< or =0.002) respectively. Furthermore, mefenamic acid reduced cerebral edema when measured as a function of brain water content. MK-801 was also neuroprotective against MCAO brain injury. This study demonstrates a significant neuroprotective effect by a fenamate NSAID against glutamate-induced cell toxicity, in vitro and against ischemic stroke in vivo. Further experiments are currently addressing the mechanism(s) of this neuroprotection. PMID- 19563852 TI - Are acute toxicity studies required to support overdose for new medicines? PMID- 19563850 TI - BDNF downregulates 5-HT(2A) receptor protein levels in hippocampal cultures. AB - Both brain-derived neurotrophic factor (BDNF) and the serotonin receptor 2A (5 HT(2A)) have been related to depression pathology. Specific 5-HT(2A) receptor changes seen in BDNF conditional mutant mice suggest that BDNF regulates the 5 HT(2A) receptor level. Here we show a direct effect of BDNF on 5-HT(2A) receptor protein levels in primary hippocampal neuronal and mature hippocampal organotypic cultures exposed to different BDNF concentrations for either 1, 3, 5 or 7 days. In vivo effects of BDNF on hippocampal 5-HT(2A) receptor levels were further corroborated in (BDNF +/-) mice with reduced BDNF levels. In primary neuronal cultures, 7 days exposure to 25 and 50ng/mL BDNF resulted in downregulation of 5 HT(2A), but not of 5-HT(1A), receptor protein levels. The BDNF-associated downregulation of 5-HT(2A) receptor levels was also observed in mature hippocampal organotypic cultures, excluding confounding effects of BDNF on immature tissue. BDNF +/- mice showed significant increased 5-HT(2A) receptor levels in hippocampus confirming the association between 5-HT(2A) receptor and BDNF levels in vivo. In conclusion, our results point to a regulatory role of BDNF on 5-HT2A receptor levels. This interaction may be an important mechanism in the role of BDNF in affective disorders emphasizing the need for further elucidating the specificity and the mechanism behind this regulation. PMID- 19563853 TI - Isolation rearing induces social and emotional function abnormalities and alters glutamate and neurodevelopment-related gene expression in rats. AB - Isolation rearing induces various cognitive abnormalities such as reversal learning deficits and reduced prepulse inhibition in rats. However, there are few reports in the literature on its effects on social and emotional functions. In the current study we aimed to address these issues and demonstrated that isolation rearing induced aggression and impaired social recognition, produced moderate anxiogenic effects in the elevated-plus maze, and resulted in hyperactivity in a novel open field. We also found NR2A, NR2B, PSD-95 and SAP-102 mRNA expression were significantly up-regulated in the hippocampus while NR2B was down-regulated in prefrontal cortex in response to isolation rearing. This study advances the use of social isolation as an animal model for studying etiological mechanisms of various neuropsychiatric disorders. PMID- 19563854 TI - Improvement of risperidone-related tardive parkinsonism with a switch to aripiprazole. PMID- 19563855 TI - Chemical composition and cytotoxic, mutagenic and genotoxic activities of the essential oil from Pipergaudichaudianum Kunth leaves. AB - We have investigated the chemical composition of Piper gaudichaudianum essential oil, as well as its cytotoxic, mutagenic and genotoxic effects in V79 cells. The chemical analyses showed that the major compounds are (E)-nerolidol (22.4%), alpha-humulene (16.5%), (E)-caryophyllene (8.9%) and bicyclogermacrene (7.4%). Dose-dependent cytotoxic effects were observed in V79 cells treated with essential oil by using clonal survival, 3-(4,5-dimethylthiazole-2-yl)-2,5 biphenyl tetrazolium bromide reduction assay (MTT) and trypan blue exclusion assay (TB), and a significant decrease in survival was observed at concentrations of 0.5 microg/mL and higher. The P. gaudichaudianum essential oil treatment caused DNA strand breaks in V79 cells at concentrations up to 2 microg/mL, as detected by the alkaline comet assay, but did not induce double-strand breaks, as verified by neutral comet assay. It induced a significant increase in the frequency of micronucleated cells at 4, 6 and 10 microg/mL. Moreover, P. gaudichaudianum essential oil significantly increased lipid peroxidation at doses of 0.5 microg/mL and higher, suggesting that the observed oxidant potential can be responsible, at least in part, for its cytotoxic and genotoxic effects. PMID- 19563856 TI - Antioxidant activities, metal contents, total phenolics and flavonoids of seven Morchella species. AB - Seven Morchella species were analyzed for their antioxidant activities in different test systems namely beta-carotene/linoleic acid, DPPH, reducing power, chelating effect and scavenging effect (%) on the stable ABTS*(+), in addition to their heavy metals, total phenolic and flavonoid contents. In beta carotene/linoleic acid system, the most active mushrooms were M. esculenta var. umbrina and M.angusticeps. In the case of DPPH, methanol extract of M. conica showed high antioxidant activity. The reducing power of the methanol extracts of mushrooms increased with concentration. Chelating capacity of the extracts was also increased with the concentration. On the other hand, in 40 microg ml(-1) concentration, methanol extract of M. conica, exhibited the highest radical scavenging activity (78.66+/-2.07%) when reacted with the ABTS*(+) radical. Amounts of seven elements (Cu, Mn, Co, Zn, Fe, Ca, and Mg) and five heavy metals (Ni, Pb, Cd, Cr, and Al) were also determined in all species. M. conica was found to have the highest phenolic content among the samples. Flavonoid content of M. rotunda was also found superior (0.59+/-0.01 microg QEs/mg extract). PMID- 19563857 TI - Evaluation of the antioxidant activity of non-transformed and transformed pineapple: a comparative study. AB - Pineapple has several beneficial properties including antioxidant activity. We investigated the antioxidant effect of different extracts of non-transformed (S) and transformed pineapple (with the magainin gene construct, [TS], for disease resistance). They were examined using 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging, oxygen radical absorbance capacity (ORAC) and lipid peroxidation assays besides phenolic and flavonoid contents. HPLC analysis was carried out to identify the possible components responsible for the differences observed. The present study indicates that the ORAC values of extracts range from 9.5 to 26.4, similar to or higher than those for some fruits and vegetables. The HPLC analysis shows that the main compounds present are ascorbic acid, quercetin, flavone-3-ols, flavones, cinnamic acids. The TS core Et. extract exhibited slightly higher concentration of ascorbic acid and considerably higher concentration of flavon-3-ols. Our study, in general, indicates that the transformation event has caused only marginal difference in antioxidant activity. Moreover the TS samples showed more antioxidant activity in some aspects and also exhibit more flavonoid content. It appears that plant cell transformation has only caused minor and favourable changes in the overall chemical composition. Thus the TS pineapple variety may have potential applications in human health like its non-transformed counterpart. PMID- 19563858 TI - Antioxidant and antilisterial effect of seed essential oil and organic extracts from Zizyphus jujuba. AB - Hydrodistilled volatile oil from the seeds of Zizyphus jujuba was analyzed by GC MS. Twenty three compounds representing 91.59% of the total oil was identified. The oil and organic extracts revealed a great potential of antilisterial effect against all five strains of Listeria monocytogenes ATCC 19111, 19116, 19118, 19166 and 15313. Also the oil had strong detrimental effect on the viable count of the tested bacteria. The samples were also subjected to screening for the antioxidant activity by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radicals scavenging activities assay. In the first case, the IC(50) value of the Z. jujuba essential oil was determined to be 5.21+/-0.01 microg/ml. Among the extracts, the strongest activity was exhibited by the methanol extract with an IC(50) value of 20.44+/-0.18 microg/ml. In the superoxide radicals scavenging activities assay, methanol extract was superior to all other extracts (IC(50)=18.60+/-0.3 microg/ml). Furthermore, the amount of total phenolic compounds was determined. The results indicate that the essential oil and extracts of Z. jujuba could serve as natural antimicrobial and antioxidant agents for the food industry. PMID- 19563859 TI - Activation of several concurrent proapoptic pathways by sulforaphane in human colon cancer cells SW620. AB - Despite the reported cytotoxicity and apoptosis-inducing properties of sulforaphane (SF) in colon cancer cells, the details concerning individual mechanisms and signaling cascades underlying SF-mediated apoptosis remain unclear. To understand different aspects of SF-induced proapoptic signaling in advanced colon carcinoma, we investigated its mechanisms in metastatic SW620 cell line. Our results indicate that in SW620 cells SF acts to induce multivariate cascades including DNA-damage response pathway whose proapoptotic signaling is nevertheless reduced owing to the mutant status of p53 and caspase-2-JNK pathway which seems to complement and enhance p53-dependent signaling, however only in wild-type p53. Furthermore, both pathways require the active role of mitochondria and do not depend on generation of ROS, making SF an attractive chemopreventive agent whose antitumor properties should be further investigated in colon cancer. PMID- 19563860 TI - Prevalence of kidney dysfunction in humans - relationship to cadmium dose, metallothionein, immunological and metabolic factors. AB - Long term cadmium (Cd) exposure in occupational and general environments may give rise to kidney dysfunction. This effect is usually considered to be the critical effect, i. e. the effect that occurs at relatively low level of exposure. The present review focused on studies of the prevalence of cadmium-related kidney dysfunction among population groups residing in cadmium contaminated areas in China. Dose-response relationships were shown between UCd and the prevalence of increased levels of biomarkers in urine of renal tubular dysfunction such as urinary beta-2-microglobulin or N-acetyl-beta-d-glucosaminidase - NAG or urinary albumin, a biomarker of glomerular kidney dysfunction. Factors that influence these dose-response relationships include: 1) Metallothionein mRNA levels in peripheral blood lymphocytes, used as a biomarker of the ability of each person, to synthesize metallothionein (a protein known to provide intracellular protection against cadmium toxicity). 2) The occurrence of increased levels in blood plasma of autoantibodies against metallothionein. 3) Concomitant changes in glucose metabolism i e Type II diabetes. 4) Concomitant exposure to other nephrotoxic agents such as inorganic arsenic. Increased susceptibility in diabetics has been shown also in population groups in Europe. In persons with type II diabetes and increased levels of autoantibodies against metallothionein in blood plasma or in persons with concomitant exposure to environmental inorganic arsenic, indications of Cd-related kidney dysfunction was observed at UCd levels around 1 microg/g creatinine, levels found among "unexposed" population groups in many countries. PMID- 19563861 TI - Synthesis, structural analysis and anticonvulsant activity of a ternary Cu(II) mononuclear complex containing 1,10-phenanthroline and the leading antiepileptic drug valproic acid. AB - The synthesis and characterization of the binary complex of copper(II) with the antiepileptic drug valproic acid sodium salt (Valp) and the related ternary complex with 1,10-phenanthroline (phen) are reported, as well as the anticonvulsant properties of the latter. The characterization was carried out by means of elemental analyses, infrared (IR), UV-visible (UV-vis) spectrophotometry and Electron Paramagnetic Resonance (EPR). The X-ray crystal structure of the mononuclear complex bis(2-propylpentanoate)(1,10-phenanthroline)copper(II) [Cu(Valp)(2)phen] is showed for the first time. It crystallized in C2/c space group with unit cell dimensions of a = 14.939(1) A, b = 19.280(1) A, c = 9.726(1) A, beta = 97.27(2) degrees , V = 2778.8(4) A(3) and Z = 8. The carboxylates bond in an asymmetric chelating mode and the copper atom adopts a highly distorted octahedral coordination, characterized by the sum of the angles of 365.9 degrees around Cu(II) and its nearest atoms in the CuN(2)O(2) + O(2) chromophore instead of the expected 360 degrees for a basal square planar geometry found in most Cu(II) complexes. Molecules assemble three by three through slipped pi-pi stacking of the aromatic phen with respectively 3.519 and 3.527 A distances, in a head-to-tail arrangement. Studies of the anticonvulsant properties of this bioligand chelate evidenced its lack of efficacy in preventing MES-induced seizures. Interestingly, complex 4 protected mice against the Minimal Clonic seizures at doses that do not cause Rotorod toxicity, with an ED(50) documenting very potent anticonvulsant activity in this model of seizure, a particularly useful pharmacological profile of activity for the treatment of Petit Mal seizures. PMID- 19563862 TI - The role of gut hormones in the regulation of body weight and energy homeostasis. AB - Obesity is one of the greatest public health challenges of the 21st century with 1.6 billion adults currently classified as being overweight and 400 million as obese. Obesity is causally associated with type 2 diabetes, hypertension, cardiovascular disease, obstructive sleep apnoea and certain forms of cancer and is now one of the leading causes of mortality and morbidity worldwide. The gastrointestinal tract is the largest endocrine organ in the body producing hormones that have important sensing and signaling roles in regulating body weight and energy expenditure. The last decade has witnessed a marked increase in our understanding of the role of gut hormones in energy homeostasis. Consequently, strategies aimed at modulating circulating gut hormone concentrations or targeting their receptors are being developed as potential pharmacotherapies for obesity. This review summarizes the current knowledge regarding the mechanisms, sites of action and effects of the anorectic gut hormones peptide tyrosine-tyrosine (PYY), pancreatic polypeptide (PP), oxyntomodulin, and amylin and of the unique orexigenic hormone, ghrelin. PMID- 19563863 TI - Mitochondrial dysfunction in CA1 hippocampal neurons of the UBE3A deficient mouse model for Angelman syndrome. AB - Angelman syndrome (AS) is a severe neurological disorder caused by a deficiency of ubiquitin protein ligase E3A (UBE3A), but the pathophysiology of the disease remains unknown. We now report that in the brains of AS mice in which the maternal UBE3A allele is mutated (m-) and the paternal allele is potentially inactivated by imprinting (p+) (UBE3A m-?p+), the mitochondria are abnormal and exhibit a partial oxidative phosphorylation (OXPHOS) defect. Electron microscopy of the hippocampal region of the UBE3A m-?p+ mice (n=6) reveals small, dense mitochondria with altered cristae, relative to wild-type littermates (n=6) and reduced synaptic vesicle density. The specific activity of OXPHOS complex III is reduced in whole brain mitochondria in UBE3A m-?p+ (n=5) mice versus wild-type littermates (n=5). Therefore, mitochondrial dysfunction may contribute to the pathophysiology of Angelman syndrome. PMID- 19563864 TI - Invertebrate models of age-related muscle degeneration. AB - Functional and structural deterioration of muscles is an inevitable consequence of ageing in a wide variety of animal species. What underlies these changes is a complex network of interactions between the muscle-intrinsic and muscle-extrinsic factors, making it very difficult to distinguish between the cause and the consequence. Many of the genes, structures, and processes implicated in mammalian skeletal muscle ageing are preserved in invertebrate species Drosophila melanogaster and Caenorhabditis elegans. The absence in these organisms of mechanisms that promote muscle regeneration, and substantially different hormonal environment, warrant caution when extrapolating experimental data from studies conducted in invertebrates to mammalian species. The simplicity and accessibility of these models, however, offer ample opportunities for studying age-related myopathologies as well as investigating drugs and therapies to alleviate them. PMID- 19563865 TI - Degradation of proteins upon storage at near-neutral pH: indications of a proteolytic/gelatinolytic activity associated with aggregates. AB - BACKGROUND: The twin phenomena of aggregation and degradation are classically associated with protein storage. However, although aggregation has been thought to be a possible consequence of protein degradation, it has never before been proposed to be a cause of degradation. METHODS: Proteins stored under physiological conditions and electrophoresed on SDS-PAGE were examined zymographically for the presence of detergent-resistant high molecular weight (HMW) forms, and association of such HMW forms with time-correlated, seeding dependent gelatinolytic activity, under various conditions. RESULTS: Eight different proteins aggregate naturally during storage at near-neutral pH, with concomitant development of 'gelatinolytic' activity diminished greatly by storage at low temperatures, extremes of pH, arginine, imidazole, BSA, azide, EDTA, DTT, PMSF (but not AEBSF), and diisopropyl fluorophosphate (DFP), suggesting involvement of surface serine residues in a novel aggregate-borne proteolytic activity. CONCLUSIONS: Naturally-formed aggregates of proteins appear to use surface serines to perform peptide bond hydrolysis, explaining degradation of proteins during storage, and indicating why aggregates are cytotoxic. GENERAL SIGNIFICANCE: The study suggests that a bi-directional cause-effect relationship operates between protein aggregation, and protein degradation, providing clues to the design of better conditions for long-term protein storage. PMID- 19563867 TI - Occurrence of Can-SINEs and intron sequence evolution supports robust phylogeny of pinniped carnivores and their terrestrial relatives. AB - Investigating the dog genome we found 178965 introns with a moderate length of 200-1000 bp. A screening of these sequences against 23 different repeat libraries to find insertions of short interspersed elements (SINEs) detected 45276 SINEs. Virtually all of these SINEs (98%) belong to the tRNA-derived Can-SINE family. Can-SINEs arose about 55 million years ago before Carnivora split into two basal groups, the Caniformia (dog-like carnivores) and the Feliformia (cat-like carnivores). Genome comparisons of dog and cat recovered 506 putatively informative SINE loci for caniformian phylogeny. In this study we show how to use such genome information of model organisms to research the phylogeny of related non-model species of interest. Investigating a dataset including representatives of all major caniformian lineages, we analysed 24 randomly chosen loci for 22 taxa. All loci were amplifiable and revealed 17 parsimony-informative SINE insertions. The screening for informative SINE insertions yields a large amount of sequence information, in particular of introns, which contain reliable phylogenetic information as well. A phylogenetic analysis of intron- and SINE sequence data provided a statistically robust phylogeny which is congruent with the absence/presence pattern of our SINE markers. This phylogeny strongly supports a sistergroup relationship of Musteloidea and Pinnipedia. Within Pinnipedia, we see strong support from bootstrapping and the presence of a SINE insertion for a sistergroup relationship of the walrus with the Otariidae. PMID- 19563866 TI - Identification of NFAT binding sites that mediate stimulation of cathepsin K promoter activity by RANK ligand. AB - The receptor activator of NFkappaB ligand (RANKL) is a critical mediator of osteoclastogenesis and regulates cathepsin K (CTSK) expression, which is essential for normal bone resorption. RANKL acts, in part, via the Ca(2+)/calmodulin/calcineurin signaling pathway, which in turn, activates NFATc1 (nuclear factor of activated T-cells) and downstream gene expression. We investigated the signals and promoter elements that regulate CTSK gene expression in RAW 264.7 cells, which can be differentiated to osteoclasts by RANKL. Disrupting Ca(2+) signaling, by blocking Ca(2+) channels, thus inhibiting calcineurin or chelation of intracellular Ca(2+), prevented the stimulation of CTSK expression by RANKL. Both RANKL treatment and overexpression of NFATc1 dramatically enhanced CTSK promoter activity, but not in an identical manner. NFATc1 regulates CTSK promoter activity, but the motifs have not been explicitly identified. We found that as few as 238 bp of the CTSK promoter were sufficient to elicit a marked response to both RANKL and NFATc1, truncations of the CTSK promoter illustrated differences in regional responsiveness. Transfection analysis of CTSK promoter-luciferase plasmids revealed that NFATc1 binding sites at 85, 289 and 345 bp upstream of the transcriptional start site mediated responses to RANKL and NFATc1. Deletion of a 4-bp core element from the site at 85 bp dramatically reduced the response of the CTSK promoter to both RANKL and NFATc1, whereas a similar deletion at -345 bp decreased NFATc1- but not RANKL mediated responses. Mutation of the site at -289 bp did not affect NFAT-mediated stimulation of CTSK on its own, but did decrease responsiveness in combination with either or both of the other two deletions. Electrophoretic mobility shift assays demonstrated NFATc1 binding to oligonucleotides containing the -85-bp and 345-bp sites, while chromatin immunoprecipitation assays demonstrated enhanced in situ binding by NFATc1 to two analogous sites in the mouse CTSK promoter in response to RANKL treatment. Therefore, proximal NFAT binding sites play a significant role in the NFATc1-mediated stimulation of CTSK gene expression by RANKL. PMID- 19563868 TI - Hypervariable 3' UTR region of plant LTR-retrotransposons as a source of novel satellite repeats. AB - The repetitive sequence PisTR-A has an unusual organization in the pea (Pisum sativum) genome, being present both as short dispersed repeats as well as long arrays of tandemly arranged satellite DNA. Cloning, sequencing and FISH analysis of both PisTR-A variants revealed that the former occurs in the genome embedded within the sequence of Ty3/gypsy-like Ogre elements, whereas the latter forms homogenized arrays of satellite repeats at several genomic loci. The Ogre elements carry the PisTR-A sequences in their 3' untranslated region (UTR) separating the gag-pol region from the 3' LTR. This region was found to be highly variable among pea Ogre elements, and includes a number of other tandem repeats along with or instead of PisTR-A. Bioinformatic analysis of LTR-retrotransposons mined from available plant genomic sequence data revealed that the frequent occurrence of variable tandem repeats within 3' UTRs is a typical feature of the Tat lineage of plant retrotransposons. Comparison of these repeats to known plant satellite sequences uncovered two other instances of satellites with sequence similarity to a Tat-like retrotransposon 3' UTR regions. These observations suggest that some retrotransposons may significantly contribute to satellite DNA evolution by generating a library of short repeat arrays that can subsequently be dispersed through the genome and eventually further amplified and homogenized into novel satellite repeats. PMID- 19563869 TI - Biomarkers of nucleic acid oxidation, polymorphism in, and expression of, hOGG1 gene in styrene-exposed workers. AB - This study investigated nucleic acid oxidation associated with styrene exposure, mRNA expression levels of hOGG1 gene and the role of the genetic polymorphism Ser326Cys of human 8-oxoguanine DNA N-glycosylase 1 (hOGG1) in 60 styrene-exposed workers and 50 unexposed clerks. Biomarkers of exposure (styrene in blood, mandelic and phenylglyoxylic acids and 4-vinylphenol in urine) and urinary biomarkers of nucleic acid oxidation, namely 8-oxo-7,8-dihydro-2'-deoxyguanosine (U-8-oxodGuo), 8-oxo-7,8-dihydroguanosine (U-8-oxoGuo) and 8-oxo-7,8 dihydroguanine (U-8-oxoGua) were determined by liquid chromatography-tandem mass spectrometry. The levels of 8-oxodGuo adduct and 2'-deoxyguanosine (dGuo) were measured by HPLC in DNA from white blood cells (WBC). Genomic DNA and RNA from blood samples were used to characterize the Ser326Cys polymorphism and the mRNA expression levels of the hOGG1 gene, respectively, by PCR-based methods. Exposed workers showed lower values of 8-oxodGuo/10(5) dGuo ratio in WBC-DNA but higher concentrations of U-8-oxoGuo compared to controls (p=0.002 and p=0.008, respectively, t-test for independent samples). In the whole group, all urinary biomarkers of nucleic acid oxidation correlated with both the sum of mandelic and phenylglyoxylic acids (rho>0.33, p<0.0001) and 4-vinylphenol (rho>0.29, p<0.001), whereas 8-oxodGuo/10(5) dGuo in WBC showed a negative correlation with exposure parameters (rho<-0.24, p<0.02). Subjects bearing the hOGG1 Ser/Ser genotype showed lower values of 8-oxodGuo/10(5) dGuo in WBC than those with at least one variant Cys allele (0.34+/-0.16 vs 0.45+/-0.21, p=0.008). In the subgroup of hOGG1 Ser/Ser subjects, laminators showed lower levels of WBC 8-oxodGuo/10(5) dGuo ratio and significantly higher concentrations of U-8-oxoGua than controls (p=0.07 and p=0.01, respectively, t-test for independent samples). Interestingly, workers showed higher levels of hOGG1 expression compared to controls (p<0.0005). Styrene exposure seems to be associated with oxidation damage to nucleic acids, particularly to RNA and with an induction of the BER system. PMID- 19563870 TI - Transdermal permeation of selegiline from hydrogel-membrane drug delivery systems. AB - In the present work, we attempted to design a transdermal system for delivering selegiline using a hydrogel-based drug reservoir and a rate-controlling membrane (Solupor polyethylene membranes). The appearances of these preparations were evaluated by scanning electron microscopy (SEM), and the in vitro skin permeation of selegiline across porcine skin was examined. Both the R- and S-forms of selegiline were examined in this study to elucidate the stereoselectivity of skin to selegiline. Solupor membranes and hydrogels exhibited a cross-linking structure with micropores. R-Selegiline revealed a flux of 1.13 microg/cm(2)/h across porcine skin. Solupor membranes were rate limiting for skin permeation of selegiline. Around a 2-fold reduction in the drug flux was determined after Solupor membrane incorporation. There were no significant differences in drug flux across the four Solupor membranes tested. The flux of R-selegiline from cellulose hydrogels approximated that from the aqueous solution (control). Both the membrane and hydrogel greatly reduced the inter-subject variations in skin permeation. According to the results of skin permeation and the partition coefficient between the skin and water (logP(skin/water)), the S-enantiomer may be preferable for permeation into the skin. However, the R- and S-forms demonstrated equal absorption of the drug fluxed in the presence of the membrane and/or the hydrogel. The results of this study encouraged us to further investigate hydrogel-membrane delivery systems for transdermal selegiline administration. PMID- 19563871 TI - Cellular automata model for swelling-controlled drug release. AB - A cellular automata approach for modeling swelling-controlled drug release is presented. In the model, a drug release device is divided into a square grid space. Each cell in the grid contains information about the material, drug, polymer or solvent in that domain. Cells are allowed to change their state according to statistical rules designed to mimic physical phenomena. Diffusion and swelling are modeled by a random walk of mobile cells, and kinetics of chemical or physical processes by probabilities of conversion from one state to another. The model is applied to drug release from a swelling binary polymer/drug device. The effect of simulation parameters on the drug release profiles and the locations of erosion and diffusion fronts are considered. The model was able to produce realistic simulations and is proposed as a new tool for the design of controlled release devices. PMID- 19563872 TI - In vitro and in vivo antitumor activity of oridonin nanosuspension. AB - The aim of the present study was to evaluate the antitumor activity of an oridonin (ORI) nanosuspension relative to ORI solution both in vitro and in vivo. ORI nanosuspension with a particle size of 897.2+/-14.2 nm was prepared by the high pressure homogenization method (HPH). MTT assay showed that ORI nanosuspension could significantly enhance the in vitro cytotoxicity against K562 cells compared to the ORI solution, the IC(50) value at 36 h was reduced from 12.85 micromol/L for ORI solution to 8.11 micromol/L for ORI nanosuspension. Flow cytometric analysis demonstrated that the ORI nanosuspension also induced a higher apoptotic rate in K562 cells compared to ORI solution. In vivo studies in a mouse model of sarcoma-180 solid tumors demonstrated significantly greater inhibition of tumor growth following treatment with ORI nanosuspension than ORI solution at the same dosage. The mice injected with ORI nanosuspension showed a higher reduction in tumor volume and tumor weight at the dose of 20mg/kg compared to the ORI solution (P<0.01), with the tumor inhibition rate increased from 42.49% for ORI solution to 60.23% for the ORI nanosuspension. Taken together, these results suggest that the delivery of ORI in nanosuspension is a promising approach for the treatment of the tumor. PMID- 19563873 TI - Design and evaluation of oral bioadhesive controlled release formulations of miglitol, intended for prolonged inhibition of intestinal alpha-glucosidases and enhancement of plasma glucagon like peptide-1 levels. AB - Alpha-glucosidase enzyme is present ubiquitously throughout the lumen of the small intestine. It is responsible for the breakdown of complex into simple carbohydrates. alpha-Glucosidase inhibitors such as miglitol, are drugs that have greater affinity towards this enzyme in comparison to carbohydrates. Miglitol regulates the postprandial glucose levels directly by inhibiting the enzyme reversibly and also indirectly by including the secretion of glucagon like peptide-1 (GLP-1). The aims of this study were (i) to design a controlled release (CR) mucoadhesive (in the intestine) formulation of miglitol which would inhibit the alpha-glucosidase enzyme for a longer duration of time (in comparison to the non-controlled release (IR) formulation) thus reducing the dosing frequency, and also controlling the postprandial glucose levels more effectively over a longer period of time; (ii) to assess the effect of different formulation parameters on the release of miglitol in vitro from the CR pellets; (iii) to evaluate the mucoadhesion of pellets in the intestine ex vivo; (iv) to study the effect of formulation parameters on plasma GLP-1 levels; and (v) to find out the effect of formulations on postprandial glucose levels. The data obtained was analysed to find out whether there was a correlation between these different parameters. Four controlled release formulations (CR1, CR2, CR3 and CR4) of miglitol comprising of multilayered pellets were designed successfully. The CR4 formulation containing 30% of 20 cps of ethyl cellulose (the retarding layer of the formulation) displayed slowest release of miglitol in vitro in comparison to other formulations. We designed an ex vivo experimental setup for studying the mucoadhesion of the pellets in the lumen of the intestine. Results indicated that amongst all of the adherent pellets, 5% were found to be adhering in the duodenal region, 61% in the jejunum, 32% in the ileum and 2% in the colon. Two of the controlled release formulations CR1 and CR4 were evaluated in vivo in dogs. Both the formulations displayed significantly higher and more prolonged (greater AUC) levels of GLP-1 in comparison to either the placebo or the immediate release (IR) formulations. They even displayed a significantly better control of postprandial glucose in comparison to either placebo or IR formulations. However, a comparison between the two controlled release formulations (CR1 and CR4) revealed that the plasma GLP-1 (AUC by CR1=63.1+/-1.32 and CR4=66.2+/-0.82) and postprandial glucose values due to both the formulations were rather similar despite their differences in in vitro release as well as pharmacokinetic profiles (plasma miglitol AUC of CR1=16.17+/-4.11 and CR4=27.17+/-4.33). PMID- 19563874 TI - Lateralized processes constrain auditory reinforcement in human newborns. AB - We investigated operant sucking response learning in human newborns. Auditory reinforcers always occurred monaurally to see whether their potency differed between ears. Experiment 1 - we controlled the reinforcers, either intrauterine heartbeat sounds or unfamiliar speech, while infants chose which ear received it. Experiment 2 - we controlled the reinforcers and the receiving ear. Unfamiliar speech reinforced learning only if infants could use their right ear and heartbeats reinforced learning only if infants could use their left ear. Experiment 3 - we controlled the ear while infants chose between their mothers' vs. a stranger's voice and between their mothers' vs. a foreign language. The more familiar speech reinforced learning only if infants could use their left ear. We proposed reinforcers' potencies differed between ears because the newborn's auditory system, just like adult's, optimizes their perceptual clarity by left-lateralized processing of their rapid temporal variations and right lateralized processing of their longer-lasting spectral characteristics. PMID- 19563875 TI - Phytochemistry and biological activities of Phlomis species. AB - The genus Phlomis L. belongs to the Lamiaceae family and encompasses 100 species native to Turkey, North Africa, Europe and Asia. It is a popular herbal tea enjoyed for its taste and aroma. Phlomis species are used to treat various conditions such as diabetes, gastric ulcer, hemorrhoids, inflammation, and wounds. This review aims to summarize recent research on the phytochemistry and pharmacological properties of the genus Phlomis, with particular emphasis on its ethnobotanical uses. The essential oil of Phomis is composed of four chemotypes dominated by monoterpenes (alpha-pinene, limonene and linalool), sesquiterpenes (germacrene D and beta-caryophyllene), aliphalic compounds (9,12,15 octadecatrienoic acid methyl ester), fatty acids (hexadecanoic acid) and other components (trans-phytol, 9,12,15-octadecatrien-1-ol). Flavonoids, iridoids and phenylethyl alcohol constitute the main compounds isolated from Phlomis extracts. The pharmacological activities of some Phlomis species have been investigated. They are described according to antidiabetic, antinociceptive, antiulcerogenic, protection of the vascular system, anti-inflammatory, antiallergic, anticancer, antimicrobial and antioxidant properties. PMID- 19563876 TI - Traditional uses of plants in the Eastern Riviera (Liguria, Italy). AB - AIM OF THE STUDY: The aim of the present study was to explore the traditional plant knowledge of a zone of the Ligurian coast, known as Riviera spezzina (RS), Eastern Liguria, Italy. MATERIALS AND METHODS: Ethnobotanical information was obtained through open informal interviews. The informants were residents, belonging to families living in the study area since generations, and involved, at least partially, in agriculture. RESULTS AND CONCLUSIONS: A total of 120 botanical taxa were recorded. Ethnobotanical data included medicinal (40.4%), alimentary (46.5%), veterinary (4.6%), domestic and cosmetic (4.3%), magic and ritual (1.8%) and other uses (2.4%). Medicinal plants are used to treat a wide range of diseases, mainly disorders of digestive system, skin, and genital urinary and respiratory traits. The key role played by undomesticated edible herbs, locally known as "erbette", in traditional cooking recipes of RS was shown. These botanicals are rich in bioactive compounds, such as polyphenols, vitamins, complex sugars, essential fatty acids and fibers, and therefore might offer a valid protection against pathological conditions. Plant uses by RS people were compared with previous ethnobotanical literature, concerning the neighbouring areas of Liguria. Our results highlight the role of the traditional use of plants in the maintenance of health and the prevention of chronic and age related diseases. PMID- 19563879 TI - Age-and caste-dependent decrease in expression of genes maintaining DNA and RNA quality and mitochondrial integrity in the honeybee wing muscle. AB - I report here an investigation of the age- and caste-specific expression patterns of nine honeybee orthologs of genes involved in repair of oxidative and methylation damage of DNA, and possibly RNA, in wing muscle tissue of the honeybee Apis mellifera. mRNA expression levels were measured in a comparative study of queens and ageing workers. Two of these genes, both potentially involved in repair and prevention of oxidative damage, showed higher expression in queens than workers and a distinct downregulation during the ageing trajectory in workers. These were an ortholog of mammalian NTH1 and a gene encoding a fusion protein which seems to be unique for the honeybee, consisting of one domain homologous to mammalian MTH1/Nudix/bacterial mutT and another domain homologous to the mitochondrial ribosomal protein gene S23. Orthologs of aag, apn1, msh6, ogg1, smug1 and two orthologs of human ABH/E. coli alkB, had stable expression levels during the ageing trajectory except high apn1 levels in overwintering workers. To estimate eventual age-dependent mitochondrial maintenance, batches of mitochondrial DNA from young and old workers and young queens were re-sequenced using Solexa/Illumina high-throughput sequencing. The results indicate at least a 50% reduction of intact mitochondrial fragments in foragers compared to young workers, winter bees and queens. PMID- 19563878 TI - Attenuated expression of SECIS binding protein 2 causes loss of telomeric reserve without affecting telomerase. AB - The family of selenoproteins have a broad range of functions, including protection against oxidative damage. Previous studies have shown that elevated levels of oxidative damage can induce accelerated loss of telomeric DNA during proliferation of mammalian cells. The incorporation of selenocysteine (Sec) into proteins in mammalian cells requires the Sec insertion sequence (SECIS) binding protein 2 (SBP2). Thus in the present study we have assessed the effect of knocking down the expression of SBP2 on telomere length. Following knock-down of SBP2 expression in two different human cell lines, the MSTO mesothelioma cell line ( approximately 5Kb average telomere length) and SY5Y neuroblastoma cell line (approximately 4.2Kb average telomere length), we observed a significant reduction (-0.6 to -1.1 Kb; P 10 micros up to a few minutes or even hours. These rare stochastic events correspond to the formation or dissociation of entire stems through cooperative base pairing/unpairing transitions. Such a clear separation of time scales in RNA dynamics has made it possible to implement coarse grained RNA simulations, which predict RNA folding and unfolding pathways including kinetically trapped structures on biologically relevant time scales of seconds to minutes. RNA folding simulations also enable to predict the formation of pseudoknots, that is, helices interior to loops, which mechanically restrain the relative orientations of other non-nested helices. But beyond static structural constraints, pseudoknots can also strongly affect the folding and unfolding dynamics of RNA, as the order by which successive helices are formed and dissociated can lead to topologically blocked transition intermediates. The resulting knotty dynamics can enhance the stability of RNA switches, improve the efficacy of co-transcriptional folding pathways and lead to unusual self-assembly properties of RNA. PMID- 19563895 TI - Production of recombinant protein and polyclonal mouse antiserum for ferritin from Sipuncula Phascolosoma esculenta. AB - The iron storage protein, ferritin, plays a key role in iron metabolism, but its regulation and functions in many invertebrate species are still largely unknown. In our previous work, an inducible ferritin cDNA from Phascolosoma esculenta with a full-length of 1017 bp has been cloned. In this follow-up study, the deducted ferritin protein sequence was predicted to be a polypeptide of 175 amino acids with a molecular mass of 20.1955kDa and an isoelectric point of 5.08. The cDNA sequence of P. esculenta ferritin was constructed into pET system expression system and efficiently expressed in E. coli BL21 under IPTG induction. The recombinant ferritin was detected as a 24 kDa protein by SDS-PAGE. After purification directly from the gel, the recombinant ferritin was used to immunize mice and the anti-serum was prepared. The antibody displayed a strong immunological reactivity and specificity when used in Western-blot analysis. For the first time, our work provided a set of molecular tools essential for the further studies of ferritin protein functions in P. esculenta. PMID- 19563896 TI - Antagonism of Aeromonas hydrophila by propolis and its effect on the performance of Nile tilapia, Oreochromis niloticus. AB - Propolis, a resinous substance collected by Apis mellifera bees from various plant sources and mixed with secreted beeswax, is a multifunctional material used by bees in the construction, maintenance, and protection of their hives. The collected propolis sample, from High Egypt, was dark-green with olive-odor. The minimal inhibition concentration (MIC) of propolis-ethanolic-extract, against Aeromonas hydrophila, was 80 microg Propolis-ethanolic-extract and crude propolis (1%) were added to artificial basal diet with (30% crude protein) to evaluate their efficacy on the fish growth-performance, immunostimulation and resistance to A. hydrophila. Two hundred and twenty-five Oreochromis niloticus (8 +/- 0.45 g/fish) were divided into three equal treatments (T) of triplet replicates. The fish of T(1) were fed on basal diet (control). The fish of T(2) were given the basal diet, containing propolis-ethanolic-extract. The fish of T(3) were given the basal diet containing crude propolis for 28 day. The fish were intraperitoneally challenged by A. hydrophila (0.2 x 10(7) cells ml(-1)) at the end of the feeding period and kept for 15 more days. The best growth rate and feed conversion ratio were obtained with T(2.) The increase in the average daily gain, specific growth rate and feed efficiency ratio were highly significances in T(2) followed by T(3) when compared with the control group. The HCT-level and monocyte-counts were increased (T(2)). No significant change, in the large lymphocytic-count was found among the three treatments (28-27-28%), while the neutrophil-count was significantly decreased (7%) with T(2) and increased (13.11%) with the control. A significant increase in serum lysozyme and serum bactericidal activities was found with T(2). The RLP against A. hydrophila was high with T(2) and T(3). The propolis-ethanolic-extract enhanced the growth, immunity and resistance of O. niloticus against A. hydrophila more than the crude propolis. PMID- 19563893 TI - Progestin stimulation of manganese superoxide dismutase and invasive properties in T47D human breast cancer cells. AB - Superoxide dismutase (SOD) occurs in two intracellular forms in mammals, copper zinc SOD (CuZnSOD), found in the cytoplasm, mitochondria and nucleus, and manganese superoxide dismutase (MnSOD), in mitochondria. Changes in MnSOD expression (as compared to normal cells) have been reported in several forms of cancer, and these changes have been associated with regulation of cell proliferation, cell death, and metastasis. We have found that progestins stimulate MnSOD in T47D human breast cancer cells in a time and physiological concentration-dependent manner, exhibiting specificity for progestins and inhibition by the antiprogestin RU486. Progestin stimulation occurs at the level of mRNA, protein, and enzyme activity. Cycloheximide inhibits stimulation at the mRNA level, suggesting that progestin induction of MnSOD mRNA depends on synthesis of protein. Experiments with the MEK inhibitor UO126 suggest involvement of the MAP kinase signal transduction pathway. Finally, MnSOD directed siRNA lowers progestin-stimulated MnSOD and inhibits progestin stimulation of migration and invasion, suggesting that up-regulation of MnSOD may be involved in the mechanism of progestin stimulation of invasive properties. To our knowledge, this is the first characterization of progestin stimulation of MnSOD in human breast cancer cells. PMID- 19563897 TI - Production of superoxides and nitric oxide generation in haemocytes of mussel Mytilus galloprovincialis (Lmk.) after exposure to cadmium: a possible involvement of Na(+)/H(+) exchanger in the induction of cadmium toxic effects. AB - The present study investigates cadmium (Cd) ability to enhance superoxides (O(2 )) and nitric oxide (NO) production (as nitrites) in haemocytes of mussel Mytilus galloprovincialis as well as the possible involvement of Na(+)/H(+) exchanger (NHE) in the induction of NADPH oxidase and NO synthase activity. PMA, a well known PKC-mediated NADPH oxidase as well as NO synthase stimulator was also used, in order to verify Cd effects on both O(2-) and NO generation. According to the results of the present study, micromolar concentrations of Cd (0.05, 5, 10 and 50 microM) seemed to enhance O(2-) and NO generation in haemocytes of mussels. Moreover, O(2-) and NO generation in haemocytes exposed to Cd could be enhanced by its ability to induce reactive oxygen species (ROS) but respiratory burst activation as well. Inhibition of NO synthase with 10 microM l-NAME, significantly attenuated Cd ability to enhance O(2-) production and diminished NO generation, thus leading to the suggestion that Cd toxic effects, started at concentration of 50 muM, could enhance NADPH oxidase and NO synthase stimulation in haemocytes of mussels. NHE seems to play a regulatory role in the induction of either O(2-) or NO generation in haemocytes exposed to the metal, since its inhibition with the use of 10 microM EIPA significantly decrease both O(2-) and NO production. The involvement of NHE in the induction of O(2-) and NO generation, probably via PKC-mediated NADPH oxidase and NO synthase activation, is likely to be crucial to haemocytes exposed to heavy metals, such as Cd. PMID- 19563898 TI - Edwardsiella tarda T6SS component evpP is regulated by esrB and iron, and plays essential roles in the invasion of fish. AB - Edwardsiella tarda is a gram-negative pathogen for hemorrhagic septicemia in a broad range of hosts. The type VI secretion system (T6SS) has recently been dissected in E. tarda to secrete EvpC, EvpI and a novel effector protein EvpP. In this study, sequencing and genetic alignments showed that evpP genes from different E. tarda isolates were highly similar and an evpP homolog was also found in Aeromonas hydrophila 0865 isolated from a diseased eel, suggesting the possible lateral gene transfer of evpP or the whole T6SS gene island. With reporter strains carrying gfp gene fused to the evpP promoter region, flow cytometric analysis revealed that transcription of evpP was positively regulated by either the two-component system EsrA-EsrB in E. tarda or the iron concentration in media. Compared with the parental strain, in-frame deletion of evpP in E. tarda EIB202 led to the significantly increased 50% lethal doses in zebrafish (Danio rerio) and Japanese flounder (Paralichthys olivaceus), decreased hemolytic activities, failure to adhere to mucus and reduced serum resistance, and complementation of an intact evpP gene restored these phenotypes in the evpP mutant. Investigation of infection kinetics indicated that the evpP deletion mutant was unable to proliferate in vivo, particularly in immune organs of fish. Moreover, the evpP deletion mutant exhibited incapacity to internalize in EPC cell model in vitro, demonstrating that EvpP in T6SS plays critical roles for invasion mechanism of E. tarda and merits as potential target for attenuated live vaccine construction. PMID- 19563899 TI - Identification of (L)-fucose-binding proteins from the Nile tilapia (Oreochromis niloticus L.) serum. AB - Lectins are carbohydrate-binding proteins with many biological functions including cellular recognition and innate immunity. In this study, a major l fucose-binding lectin from the serum of Nile tilapia (Oreochromis niloticus L.), designated as TFBP, was isolated by l-fucose-BSA Sepharose CL6B affinity chromatography. The SDS-PAGE (10%) analysis of TFBP revealed a major band of approximately 23 kDa with an N-terminal amino acid sequence of DQTETAGQQSXPQDIHAVLREL which did not give significant similarities to the protein databases using BLASTp searches. Ruthenium red staining indicate positive calcium binding property of TFBP. The purified TFBP agglutinated human type O erythrocytes but not the type A and B fresh erythrocytes. Live Aeromonas hydrophila and Enterococcus faecalis cells were also agglutinated by the lectin. The fucose-binding proteins were detected in the soluble protein extracts from the gills, gut, head kidneys, liver, serum and spleen using a fucose-binding protein probe (l-fucose-BSA-horseradish peroxidase). The binding of TFBP with the l-fucose-BSA probe was inhibited by l-fucose but not by alpha-methyl-d-mannose. PMID- 19563900 TI - ERP and behavioural evidence for direct suppression of unwanted memories. AB - There are some past experiences that we would prefer not to remember. Previous research has shown that repeatedly stopping retrieval of an unwanted memory increases the probability of later forgetting of that memory, and engages prefrontal control mechanisms to attenuate activity in the hippocampus. However, the mechanisms of preventing memory retrieval, and how these relate to the later forgetting, are yet to be fully understood. Here we present neural and behavioural evidence that two distinct strategies for retrieval stopping - direct memory suppression and self-distracting thought substitution - contribute to forgetting of unwanted memories in qualitatively different ways. Only direct memory suppression reduced centro-parietal positivity in the event-related potentials (ERP) between 300 and 600 ms post-stimulus, consistent with a reduction in the ERP correlate of recollection. Furthermore, only direct memory suppression produced later inhibitory forgetting that was predicted by an earlier negative ERP effect that may be associated with motor inhibition. In contrast, thought substitution produced later non-inhibitory forgetting and had no effect on the ERP correlate of recollection. Our findings demonstrate the first ERP and behavioural dissociation between inhibitory and non-inhibitory forgetting, and suggest that unwanted memories may be directly suppressed without selective retrieval of alternative memories. PMID- 19563901 TI - The Candida albicans histidine kinase Chk1p: signaling and cell wall mannan. AB - Several published functions associated with the CHK1 histidine kinase of Candida albicans resemble those of the MAPK Cek1p and its cognate receptor Sho1p (SSU81). To explore this further, we have compared mutants lacking the proteins mentioned above and have constructed a double sho1/chk1Delta null mutant to determine relationships among these proteins. We observed that the sensitivity to Congo red (CR), calcofluor white (CW), as well as clumping of cells, was slightly increased in the double mutant compared to the single chk1Delta or sho1Delta mutants. However, Cek1p phosphorylation via Sho1p, which occurs during log phase growth in the presence or absence of CR in Wt cells, does not require Chk1p. These data suggest that Chk1p and Sho1p are components of parallel but independent signal pathways. In addition, bulk mannan of strains was analyzed by GLC/MS and GPC MALLS and NMR. Compared to Wt and a CHK1 gene-reconstituted strain (CHK23) that contained high, intermediate and low Mw mannan species, we found that the mannan of strains CHK21 (chk1Delta null), the cek1Delta null, and the double mutant consisted only of low Mw mannan. The sho1Delta null mutant only demonstrated a reduced intermediate type of mannan. Alcian blue binding was lower in cek1Delta, chk1Delta, and the double sho1/chk1Delta null mutant lacking high and intermediate Mw mannan than in the sho1Delta null which had a partial loss of intermediate Mw mannan only. We conclude that the Chk1p HK is part of a functionally similar but parallel pathway to the Sho1p-Cek1p pathway that confers resistance to the cell wall inhibitors CR and CW. However, a functional relationship in mannan biosynthesis of Chk1p and Cek1p exists that only partially requires Sho1p. PMID- 19563903 TI - Characterization of hexamerin proteins and their mRNAs in the adult lubber grasshopper: The effects of nutrition and juvenile hormone on their levels. AB - The lubber grasshopper (Romalea microptera) has three major hemolymph proteins with apparent sizes on native PAGE of 90, 270, and 500kDa and subunit sizes (79, 81 and 82kDa respectively) determined by SDS-PAGE . Trypsin fragments from each protein band were sequenced, used to design degenerate primers to amplify core cDNA fragments, which were extended by 5' and 3' RACE. All three cDNAs were closely related to insect hexamerins, had an N-terminal signal sequence, and their transcripts were found solely in the fat body. Adult females fed an ad libitum diet had their highest hexamerin levels on day 18 when oocytes begin rapid growth. Hexamerin levels fell as oocytes reached their maximum length on day 30. Animals fed a restricted diet had their highest hexamerin levels on day 30 which then fell as oocytes reached their maximum length on day 36. Hexamerin mRNA levels were only modestly different for animals on the two diets, indicating that nutrition affected translation of the hexamerin mRNA. Allatectomized animals treated with juvenile hormone III (JH) or methoprene caused the appearance of vitellogenin in the hemolymph, but had no effect on hexamerin levels. Thus, JH does not appear to directly regulate hexamerin production. PMID- 19563902 TI - HdaA, a class 2 histone deacetylase of Aspergillus fumigatus, affects germination and secondary metabolite production. AB - Histone deacetylases (HDACs) play an important role in regulation of gene expression through histone modifications. Here we show that the Aspergillus fumigatus HDAC HdaA is involved in regulation of secondary metabolite production and is required for normal germination and vegetative growth. Deletion of the hdaA gene increased the production of several secondary metabolites but decreased production of gliotoxin whereas over-expression hdaA increased production of gliotoxin. RT-PCR analysis of 14 nonribosomal peptide synthases indicated HdaA regulation of up to nine of them. A mammalian cell toxicity assay indicated increased activity in the over-expression strain. Neither mutant affected virulence of the fungus as measured by macrophage engulfment of conidia or virulence in a neutropenic mouse model. PMID- 19563904 TI - Long-chain polyunsaturated fatty acid synthesis in fish: Comparative analysis of Atlantic salmon (Salmo salar L.) and Atlantic cod (Gadus morhua L.) Delta6 fatty acyl desaturase gene promoters. AB - Fish vary in the ability to biosynthesise n-3 long-chain polyunsaturated fatty acids (LC-PUFA), with marine fish such as cod being inefficient in comparison to freshwater and salmonid fish. We investigated differences in the gene promoters of Delta6 fatty acyl desaturase (Delta6 FAD), a critical enzyme in LC-PUFA biosynthesis, in cod and salmon. Progressive deletions and targeted mutations of the promoters were tested for activity in a transfected Atlantic salmon (AS) cell line under low or high LC-PUFA treatment, and regions sufficient to direct transcription were identified. Comparison of these regions with sequences of corresponding regions of Delta6 FAD genes from mammals, amphibians and fish indicated a remarkable conservation of binding sites for SREBPs and NF-Y. In addition to these sites, a site was identified in salmon with similarity to that recognised by Sp1 transcription factor, and which was required for full expression of the salmon Delta6 FAD gene. The cod promoter was less active and lacked the Sp1 site. Eicosapentaenoic acid suppressed LC-PUFA synthesis in AS cells and also suppressed activity of the salmon Delta6 FAD promoter although this activity was likely mediated through sites other than Sp1, possibly similar to those recognised by NF-Y and SREBP transcription factors. PMID- 19563905 TI - p63 in Mytilus galloprovincialis and p53 family members in the phylum Mollusca. AB - Genes of the p53 family are known to be critical regulators of the cell cycle. They have already been established as possible biomarkers. Elaborate regulation mechanisms result in numerous cDNA and protein isoforms being expressed from each gene of the p53 family. Their similarity caused an often misleading nomenclature in non-vertebrate species. The aim of the present work is a clarification of the nomenclature of molluscan p53 family sequences, an essential prerequisite for reliable interpretation of gene expression and protein function studies. Here, we report five partial cDNA and one partial genomic p63 sequences, all originating from two Mytilus galloprovincialis individuals. DNA, deduced protein sequences, and the exon/intron architecture were analyzed and compared to p53, p63 and p73 sequences from other organisms. Along with our sequences, we analyzed all similar molluscan sequences found in the GenBank database. The analysis showed our cDNA sequences code for the TAp63gamma isoform of the p63 protein, and identified all other molluscan p53 family sequences as p63 genes or their expression isoforms. Our results also indicate p63 as the ancestral gene of the p53 family as well as the only gene of the family present in non-chordate metazoan species. PMID- 19563906 TI - A novel approach for assessing protein synthesis in channel catfish, Ictalurus punctatus. AB - A comprehensive understanding of animal growth requires adequate knowledge of protein synthesis (PS), which in fish, has traditionally been determined by the flooding dose method. However, this procedure is limited to short-term assessments and may not accurately describe fish growth over extended periods of time. Since deuterium oxide ((2)H(2)O) has been used to non-invasively quantify PS in mammals over short- and long-term periods, we aimed at determining if (2)H(2)O could also be used to measure PS in channel catfish. Fish were stocked in a 40-L aquarium with approximately 4% (2)H(2)O and sampled at 4, 8 and 24h (n=6 at each time period) to determine (2)H-labeling of body water (plasma), as well as protein-free and protein-bound (2)H-labeled alanine. The labeling of body water reflected that of aquarium water and the labeling of protein-free alanine remained constant over 24h and was approximately 3.8 times greater than that of body water. By measuring (2)H-labeled alanine incorporation after 24h of (2)H(2)O exposure we were able to calculate a rate of PS: 0.04+/-0.01% h(-1). These results demonstrate that PS in fish can be effectively measured using (2)H(2)O and, because this method yields integrative measures of PS, is relatively inexpensive and accounts for perturbations such as feeding, it is a novel and practical assessment option. PMID- 19563907 TI - A Toxoplasma type 2C serine-threonine phosphatase is involved in parasite growth in the mammalian host cell. AB - Toxoplasma gondii is a human protozoan parasite that belongs to the phylum of Apicomplexa and causes toxoplasmosis. As the other members of this phylum, T. gondii obligatory multiplies within a host cell by a peculiar type of mitosis that leads to daughter cell assembly within a mother cell. Although parasite growth and virulence have been linked for years, few molecules controlling mitosis have been yet identified and they include a couple of kinases but not the counteracting phosphatases. Here, we report that in contrast to other animal cells, type 2C is by far the major type of serine threonine phosphatase activity both in extracellular and in intracellular dividing parasites. Using wild type and transgenic parasites, we characterized the 37kDa TgPP2C molecule as an abundant cytoplasmic and nuclear enzyme with activity being under tight regulation. In addition, we showed that the increase in TgPP2C activity significantly affected parasite growth by impairing cytokinesis while nuclear division still occurred. This study supports for the first time that type 2C protein phosphatase is an important regulator of cell growth in T. gondii. PMID- 19563911 TI - Could a strong alkali deproteinization replace the standard lysis step in alkaline single cell gel electrophoresis (comet) assay (pH>13)? AB - The alkaline version of single cell gel electrophoresis (comet) assay is widely used for evaluating DNA damage at the individual cell level. The standard alkaline method of the comet assay involves deproteinization of cells embedded in agarose gel using a high salt-detergent lysis buffer, followed by denaturation of DNA and electrophoresis using a strong alkali at pH>13 [N.P. Singh, M.T. McCoy, R.R. Tice, E.L. Schneider, A simple technique for quantitation of low levels of DNA damage in individual cells, Exp. Cell. Res. 175 (1988) 184-191]. However, a recent report showed that a strong alkali treatment results in simultaneous deproteinization of cells and denaturation of genomic DNA [P. Sestili, C. Martinelli, V. Stocchi, The fast halo assay: an improved method to quantify genomic DNA strand breakage at the single cell-level, Mutat. Res. 607 (2006) 205 214]. This study was carried out to test whether the strong alkali deproteinization of cells could replace the high salt-detergent lysis step used in the standard method of the alkaline comet assay. Peripheral blood lymphocytes from 3 healthy individuals were irradiated with gamma rays at doses varying between 0 and 10 Gy. Following irradiation, the comet assay was performed according to the standard alkaline method (pH>13) and a modified method. In the modified method, agarose embedded cells were treated with a strong alkali (0.3M NaOH, 0.02 M Trizma and 1mM EDTA, pH>13) for 20 min to allow deproteinization of cells and denaturation of DNA. This was followed by electrophoresis using the same alkali solution to obtain comets. DNA damage expressed in terms of comet tail length, percentage of DNA in comet tail and tail moment obtained by the standard alkaline method and the modified method were compared. In both methods, DNA damage showed a good correlation with the dose of gamma ray. The results indicate a satisfactory sensitivity of the modified method in detecting radiation induced DNA damage in human peripheral blood lymphocytes. PMID- 19563910 TI - PPARgamma activation induces autophagy in breast cancer cells. AB - It has been previously shown that PPAR gamma ligands induce apoptotic cell death in a variety of cancer cells. Given the evidence that these ligands have a receptor-independent function, we further examined the specific role of PPAR gamma activation in this biological process. Surprisingly, we failed to demonstrate that MDA-MB-231 breast cancer cells undergo apoptosis when treated with sub-saturation doses of troglitazone and rosiglitazone, which are synthetic PPAR gamma ligands. Acridine orange (AO) staining showed acidic vesicular formation within ligand-treated cells, indicative of autophagic activity. This was confirmed by autophagosome formation as indicated by redistribution of LC3, an autophagy-specific protein, and the appearance of double-membrane autophagic vacuoles by electron microscopy following exposure to ligand. To determine the mechanism by which PPAR gamma induces autophagy, we transduced primary mammary epithelial cells with a constitutively active mutant of PPAR gamma and screened gene expression associated with PPAR gamma activation by genome-wide array analysis. HIF1 alpha and BNIP3 were among 42 genes up-regulated by active PPAR gamma. Activation of PPAR gamma induced HIF1 alpha and BNIP3 protein and mRNA abundance. HIF1 alpha knockdown by shRNA abolished the autophagosome formation induced by PPAR gamma activation. In summary, our data shows a specific induction of autophagy by PPAR gamma activation in breast cancer cells providing an understanding of distinct roles of PPAR gamma in tumorigenesis. PMID- 19563912 TI - Identification and characterization of a novel 5 bp deletion in a putative insulin response element in the lipoprotein lipase gene. AB - Our aim was to identify an insulin response element (IRE) in the lipoprotein lipase (LPL) gene. We identified a 19 bp sequence as a putative IRE in LPL non coding exon 10 using bioinformatics. Upon sequencing the IRE region, a novel 5 bp deletion was identified in Hispanics (N=406) with a carrier frequency of 4.2% but not in non-Hispanic whites (N=604) or Africans. Electrophoretic mobility shift assay revealed binding sites for regulatory factor(s) in muscle cell nuclear extracts with putative IRE sequence. Antibody supershift assay using human aorta smooth muscle cell nuclear extract revealed that Elk-1 specifically binds to putative IRE. TaqMan real-time RT-PCR of the 5 bp deletion, the mutant and wild type cDNA expressed in COS-1 and human muscle cells revealed that the 5 bp deletion was associated with modest reduction in LPL expression. There was also a slight reduction in LPL translation in the deletion mutant. Our data suggest the presence of an IRE in the 3'UTR of the LPL gene. PMID- 19563908 TI - Immune escape mechanisms of intraocular tumors. AB - The notion that the immune system might control the growth of tumors was suggested over 100 years ago by the eminent microbiologist Paul Ehrlich. This concept was refined and expanded by Burnet and Thomas 50 years later with their articulation of the "immune surveillance" hypothesis. In its simplest form, the immune surveillance hypothesis suggests that neoplasms arise spontaneously and express novel antigens that are recognized by the immune system, which either eliminates the tumors or restrains their growth. Within the eye, immune responses are controlled and sometimes profoundly inhibited - a condition known as immune privilege. Immune privilege in the eye is the result of a complex array of anatomical, physiological, and immunoregulatory mechanisms that prevent the induction and expression of many immune responses. Tumors arising in the eye would seem to have an advantage in evading immune surveillance due to ocular immune privilege. Uveal melanoma, the most common and malignant intraocular tumor in adults, not only benefits from the immune privilege of the eye but also has adopted many of the mechanisms that contribute to ocular immune privilege as a strategy for protecting uveal melanoma cells once they leave the sanctuary of the eye and are disseminated systemically in the form of metastases. Although the immune system possesses a battery of effector mechanisms designed to rid the body of neoplasms, tumors are capable of rapidly evolving and countering even the most sophisticated immunological effector mechanisms. To date, tumors seem to be winning this arms race, but an increased understanding of these mechanisms should provide insights for designing immunotherapy that was envisioned over half a century ago, but has failed to materialize to date. PMID- 19563916 TI - A neonatal polyvisceral failure linked to a de novo homoplasmic mutation in the mitochondrially encoded cytochrome b gene. AB - Mutations within the mitochondrially encoded cytochrome b (MTCYB) gene are heteroplasmic and lead to severe exercise intolerance. We describe an unusual clinical presentation secondary to a novel homoplasmic mutation within MTCYB. The m.15635T>C transition (S297P) was carried by a newborn who presented with a polyvisceral failure. This mutation was responsible for a complex III deficiency. It was homoplasmic in all tissues tested and was undetectable in patient's mother. Functional analyses, including studies on patient's cybrid cell lines, demonstrate the pathogenicity of this variant. Our data show that mutations within MTCYB can be responsible for severe phenotype at birth. PMID- 19563909 TI - The neurovascular retina in retinopathy of prematurity. AB - The continuing worldwide epidemic of retinopathy of prematurity (ROP), a leading cause of childhood visual impairment, strongly motivates further research into mechanisms of the disease. Although the hallmark of ROP is abnormal retinal vasculature, a growing body of evidence supports a critical role for the neural retina in the ROP disease process. The age of onset of ROP coincides with the rapid developmental increase in rod photoreceptor outer segment length and rhodopsin content of the retina with escalation of energy demands. Using a combination of non-invasive electroretinographic (ERG), psychophysical, and image analysis procedures, the neural retina and its vasculature have been studied in prematurely born human subjects, both with and without ROP, and in rats that model the key vascular and neural parameters found in human ROP subjects. These data are compared to comprehensive numeric summaries of the neural and vascular features in normally developing human and rat retina. In rats, biochemical, anatomical, and molecular biological investigations are paired with the non invasive assessments. ROP, even if mild, primarily and persistently alters the structure and function of photoreceptors. Post-receptor neurons and retinal vasculature, which are intimately related, are also affected by ROP; conspicuous neurovascular abnormalities disappear, but subtle structural anomalies and functional deficits may persist years after clinical ROP resolves. The data from human subjects and rat models identify photoreceptor and post-receptor targets for interventions that promise improved outcomes for children at risk for ROP. PMID- 19563917 TI - Analysis of mitochondrial DNA variants in Japanese patients with schizophrenia. AB - To test the hypothesis that mitochondrial DNA (mtDNA) variants contribute to the susceptibility to schizophrenia, we sequenced the entire mtDNAs from 93 Japanese schizophrenic patients. Three non-synonymous homoplasmic variants in subunit six of the ATP synthase (MT-ATP6) gene that were detected only in patients but not in controls were suggested to be slightly deleterious, because (1) their original amino acid residues (AA) were highly conserved and (2) the physicochemical differences between the original and altered AA were relatively high. In addition, we detected three novel heteroplasmic variants that were potentially pathogenic. Although functional analysis is needed, rare variants in the mtDNA may convey susceptibility to schizophrenia. PMID- 19563919 TI - Tracing metabolic pathways from enzyme data. AB - The IUBMB Enzyme List is widely used by other databases as a source for avoiding ambiguity in the recognition of enzymes as catalytic entities. However, it was never designed for activities such as pathway tracing, which have become increasingly important in systems biology. This is because it often relies on generic or representative reactions to show the reactions catalysed by enzymes of wide specificity. It is necessary to go to databases such as BRENDA to find further, more detailed, information on what is known about the range of substrates for any particular enzyme. In order to provide a framework for tracing pathways involving any specific enzyme or metabolite, we have created a Reactions Database from the material in the Enzyme List. This allows reactions to be searched by substrate/product and pathways to be traced from any selected starting/seed substrate. An extensive synonym glossary allows searches by many of the alternative names, including accepted abbreviations, by which a chemical compound may be known. This database was necessary for the development of the application Reaction Explorer (http://www.reaction-explorer.org), which was written in REALbasic to search the Reactions Database and draw metabolic pathways from reactions selected by the user. Having input the name of the starting compound (the "seed"), the user is presented with a list of all reactions containing that compound and then selects the product of interest as the next point on the ensuing graph. The pathway diagram is then generated as the process iterates. A contextual menu is provided, which allows the user to (i) remove a compound from the graph, along with all associated links; (ii) search the reactions database again for additional reactions involving the compound and (iii) search for the compound within the Enzyme List. PMID- 19563918 TI - What are the future top priority questions in cardiovascular research and what new hardware needs to be developed? AB - The final discussion that took place at the Cardiovascular Workshop in Strasbourg in November 2008 identified the following priorities for our future research and the need to develop new dedicated cardiovascular devices. Knowledge and treatment of cardiovascular deconditioning remain limited. The prospect of very long duration space flights could lead to the emergence of new cardiovascular risks. We need to maintain our efforts to standardize our experimental protocols and analysis equipment. We continue to rely on animal models for cardiovascular research. To continue our studies we recommend developing specific tools for the space environment so that they can be made available to the scientific and medical community. PMID- 19563915 TI - Mitochondrial kinases in Parkinson's disease: converging insights from neurotoxin and genetic models. AB - Alterations in mitochondrial biology have long been implicated in neurotoxin, and more recently, genetic models of parkinsonian neurodegeneration. In particular, kinase regulation of mitochondrial dynamics and turnover are emerging as central mechanisms at the convergence of neurotoxin, environmental and genetic approaches to studying Parkinson's disease (PD). Kinases that localize to mitochondria during neuronal injury include mitogen activated protein kinases (MAPK) such as extracellular signal regulated protein kinases (ERK) and c-Jun N-terminal kinases (JNK), protein kinase B/Akt, and PTEN-induced kinase 1 (PINK1). Although site(s) of action within mitochondria and specific kinase targets are still unclear, these signaling pathways regulate mitochondrial respiration, transport, fission fusion, calcium buffering, reactive oxygen species (ROS) production, mitochondrial autophagy and apoptotic cell death. In this review, we summarize accelerating experimental evidence gathered over the last decade that implicate a central role for kinase signaling at the mitochondrion in Parkinson's and related neurodegenerative disorders. Interactions involving alpha-synuclein, leucine rich repeat kinase 2 (LRRK2), DJ-1 and Parkin are discussed. Converging mechanisms from different model systems support the concept of common pathways in parkinsonian neurodegeneration that may be amenable to future therapeutic interventions. PMID- 19563920 TI - Leishmania donovani trypanothione reductase: role of urea and guanidine hydrochloride in modulation of functional and structural properties. AB - Trypanothione reductase [TR], an NADPH-dependent disulfide oxidoreductase, unique to kinetoplastid parasites including Trypanosoma and Leishmania, is a validated target for the design of improved drugs. TR is a stable homodimer with a FAD molecule tightly bound to each subunit. In this paper, structure, function, stability properties and cofactor protein interactions of recombinant TR from Leishmania donovani were investigated under equilibrium unfolding/denaturing conditions. Urea induced unfolding was non-reductive in nature and led to the formation of partially folded intermediate. This intermediate species lacks catalytic activity and characteristic conformation of native LdTR but has significant secondary structure and could be partially reactivated. Guanidine hydrochloride-induced irreversible denaturation was marked by the presence of molten globule intermediate. Reactivation and cross-linking experiments clearly demonstrated that the loss of activity at lower denaturant concentrations was not coincided by dimer dissociation or structural unfolding. The studies demonstrate that functional conformation and stability are largely governed by ionic interactions and active site disulfide plays a vital role in maintaining functional conformation. The results obtained from this study provide intriguing insight into the possible mechanism/s of modulation of structure, function and stability of LdTR induced by the cationic, guanidine hydrochloride and the neutral denaturant, urea. PMID- 19563921 TI - Proteomic characterization of the dynamic KSR-2 interactome, a signaling scaffold complex in MAPK pathway. AB - KSR-1 is a scaffold protein that is essential for Ras-induced activation of the highly conserved RAF-MEK-ERK kinase module. Previously, we identified a close homolog of KSR-1, called KSR-2, through structural homology-based data mining. In order to further understand the role of KSR-2 in MAPK signaling, we undertook a functional proteomics approach to elucidate the dynamic composition of the KSR-2 functional complex in HEK-293 cells under conditions with and without TNF-alpha stimulation. We found nearly 100 proteins that were potentially associated with KSR-2 complex and 43 proteins that were likely recruited to the super molecular complex after TNF-alpha treatment. Our results indicate that KSR-2 may act as a scaffold protein similar as KSR-1 to mediate the MAPK core (RAF-MEK-ERK) signaling but with a distinct RAF isoform specificity, namely KSR-2 may only mediate the A-RAF signaling while KSR-1 is responsible for transducing signals only from c-RAF. In addition, KSR-2 may be involved in the activation of many MAPK downstream signaling molecules such as p38 MAPK, IKAP, AIF, and proteins involved in ubiquitin-proteasome, apoptosis, cell cycle control, and DNA synthesis and repair pathways, as well as mediating crosstalks between MAPK and several other signaling pathways, including PI3K and insulin signaling. While interactions with these molecules are not known for KSR-1, it's reasonable to hypothesize that KSR-1 may also play a similar role in mediating these downstream signaling pathways. PMID- 19563922 TI - Impact of co-incorporating laminin peptide dopants and neurotrophic growth factors on conducting polymer properties. AB - Conductive neural interfaces tailored for cell interaction by incorporation of bioactive factors are hypothesized to produce superior neuroprostheses with improved charge transfer capabilities. This study examined the effect of entrapping nerve growth factor (NGF) within the conducting polymer poly(ethylene dioxythiophene) (PEDOT) during electrodeposition to create a polymer capable of stimulating neurite outgrowth from proximal neural tissue. NGF entrapment was performed on polymers doped with laminin peptides DEDEDYFQRYLI and DCDPGYIGSR and, additionally, a conventional dopant, paratoluene sulphonate (pTS). All polymer coatings were analysed for a range of physical, electrical and mechanical properties, with the biological activity of ligands examined using a PC12 neurite outgrowth assay. NGF was successfully entrapped in PEDOT during electrodeposition and was shown to produce a softer interface than conventional conducting polymers and films without the NGF modification. However, it was found that the use of a peptide dopant combined with NGF entrapment resulted in polymers with diminished electrical and mechanical stability. Entrapped NGF was determined to be biologically active, with PEDOT/pTS/NGF producing neurite outgrowth comparable with control films where NGF was supplied via the medium. Future studies will determine the effect of typical neural prosthetic stimulation regimes on the release of neurotrophins and subsequent cell response. PMID- 19563923 TI - Direct write assembly of calcium phosphate scaffolds using a water-based hydrogel. AB - The development of materials to support bone regeneration requires flexible fabrication technologies able to tailor chemistry and architecture for specific applications. In this work we describe the preparation of ceramic-based inks for robotic-assisted deposition (robocasting) using Pluronic F-127 solutions. This approach allows the preparation of pseudoplastic inks with solid contents ranging between 30 and 50 vol.%, enabling them to flow through a narrow printing nozzle while supporting the weight of the printed structure. Ink formulation does not require manipulation of the pH or the use of highly volatile organic components. Therefore, the approach can be used to prepare materials with a wide range of compositions, and here we use it to build hydroxyapatite (HA), beta-tricalcium phosphate (beta-TCP) and biphasic (HA/beta-TCP) structures. The flow of the inks is controlled by the Pluronic content and the particle size distribution of the ceramic powders. The use of wide size distributions favors flow through the narrow printing nozzles and we have been able to use printing nozzles as narrow as 100 microm in diameter, applying relatively low printing pressures. The microporosity of the printed lines increases with increasing Pluronic content and lower sintering temperatures. Microporosity can play a key role in determining the biological response to the materials, but it also affects the strength of the structure. PMID- 19563925 TI - Lateral bone density variations in the scoliotic spine. AB - Adolescent Idiopathic Scoliosis (AIS) is the most common deformity of the spine, affecting 2-4% of the population. Previous studies have shown that the vertebrae in scoliotic spines undergo abnormal shape changes, however there has been little exploration of how scoliosis affects bone density distribution within the vertebrae. In this study, existing CT scans of 53 female idiopathic scoliosis patients with right-sided main thoracic curves were used to measure the lateral (right to left) bone density profile at mid-height through each vertebral body. Five key bone density profile measures were identified from each normalized bone density distribution, and multiple regression analysis was performed to explore the relationship between bone density distribution and patient demographics (age, height, weight, body mass index (BMI), skeletal maturity, time since Menarche, vertebral level, and scoliosis curve severity). Results showed a marked convex/concave asymmetry in bone density for vertebral levels at or near the apex of the scoliotic curve. At the apical vertebra, mean bone density at the left side (concave) cortical shell was 23.5% higher than for the right (convex) cortical shell, and cancellous bone density along the central 60% of the lateral path from convex to concave increased by 13.8%. The centre of mass of the bone density profile at the thoracic curve apex was located 53.8% of the distance along the lateral path, indicating a shift of nearly 4% toward the concavity of the deformity. These lateral bone density gradients tapered off when moving away from the apical vertebra. Multi-linear regressions showed that the right cortical shell peak bone density is significantly correlated with skeletal maturity, with each Risser increment corresponding to an increase in mineral equivalent bone density of 4-5%. There were also statistically significant relationships between patient height, weight and BMI, and the gradient of cancellous bone density along the central 60% of the lateral path. Bone density gradient is positively correlated with weight, and negatively correlated with height and BMI, such that at the apical vertebra, a unit decrease in BMI corresponds to an almost 100% increase in bone density gradient. PMID- 19563926 TI - Short-term relationship between meteorological variables and hip fractures: an analysis carried out in a health area of the Autonomous Region of Valencia, Spain (1996-2005). AB - INTRODUCTION: Diverse studies have shown a seasonal influence on the incidence of hip fracture (HF), possibly associated with adverse meteorological conditions. In this paper, we present an analysis of the short-term relationship between meteorological conditions and the incidence of HF in people over 45 years of age living in a Mediterranean climate zone. MATERIAL AND METHODS: HF cases admitted to the two reference hospitals in the health area were selected. The meteorological variables (temperature, relative humidity, rain, wind, and other conditions) were obtained from a weather station centrally located within the area under study. The trend and seasonality of the time series were analyzed with the aid of Poisson regression modeling. The relationship between the incidence of a hip fracture and the meteorological conditions, both on the same day and on the day previous to the patient's admission to the hospital were correlated in a case crossover analysis with the control periods selected in accordance with two different methods of approximation (symmetric and semi-metric). The results were analyzed for different subgroups defined by age (older or younger than 75 years of age) and sex of subject and by type of fracture (cervical or pertrochanteric). RESULTS: 2121 patients admitted for HF were selected. Of these, 1598 (75.3%) were women and 523 (24.7%) were men. The average age of the subjects was 80, with patients ranging from 45-99 years of age. The time/weather series showed a positive tendency, with a greater occurrence of cases in the autumn and winter months. The case-crossover analysis showed a significant relationship between the daily duration of wind and the incidence of HFs. Divided into quartiles, the windiest days (quartile 4) were associated with a 32% increased risk of HF (OR 1.32 CI 95% 1.10-1.58) with respect to the calmest days (quartile 1), especially in patients under 75 (OR 1.53; CI 95% 1.02-2.29). The remaining meteorological variables were not associated in any significant fashion with the incidence of HFs. The results were comparable across different subgroups classified by age, sex, and type of fracture. CONCLUSIONS: The incidence of HFs varies seasonally and presents a significant association with the coldest times of the year. In the short-term, lasting windy periods are associated with an increased risk of HFs whereas other meteorological variables show a much lower, non-significant correlation. PMID- 19563924 TI - Longitudinal changes of serum COMP and urinary CTX-II predict X-ray defined knee osteoarthritis severity and stiffness in women. AB - OBJECTIVE: To ascertain the predictive role of longitudinally acquired biochemical measures of cartilage turnover in relation to X-ray defined knee osteoarthritis (OAK), knee pain and functioning. METHODS: This is a feasibility study based on 72 enrollees of the Michigan site of Study of Women's Health Across the Nation (SWAN), a longitudinal, population-based cohort study with 11 annual visits to characterize health at the mid-life. At visits in 1996, 1998 and 2007, radiographs were evaluated for the presence of OAK [>or=2 on the Kellgren and Lawrence (K-L) scale]. Knee pain and stiffness were assessed by interview. Functioning was assessed using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Cartilage oligomeric matrix protein (COMP) and Type II collagen telopeptides (CTX-II) were assayed in serum and urine samples collected on alternate years from 1997 to 2006. We related trajectories of the cartilage biochemical markers from these five time points to OAK severity (no OAK, K-L score<2; mild OAK, K-L score=2; moderate/severe OAK, K-L score=3 or 4), pain, stiffness, or functioning, using longitudinal non-linear mixed modeling. RESULTS: The 2007 prevalence of X-ray defined OAK was 50% in these 72 women. Upward trajectories of COMP (P=0.02) and CTX-II (P=0.006) were associated with increased OAK severity and body size. COMP trajectories were associated with pain and stiffness, but not functioning. CTX-II trajectories were associated with stiffness scores, but not knee pain or functioning scores. CONCLUSION: Multiple, biennial measures of COMP or CTX-II taken over a 10-year period were predictive of subsequent OAK and knee stiffness. PMID- 19563927 TI - Polymorphisms of cytochrome P450 1A1, cigarette smoking and risk of coronary artery disease. AB - Epidemiological evidence has demonstrated a strong relationship between cigarette smoking and coronary artery disease (CAD). Cytochrome P450 1A1 (CYP1A1) is a key enzyme that metabolizes the cigarette toxin relevant to smoking-induced atherogenesis. This case-control study examined the role of CYP1A1 polymorphisms, CYP1A1 2A (T6235C) and CYP1A1 2C (A4889G), in susceptibility to smoking-related CAD. We recruited 481 patients with 50% or more luminal obstructions in the coronary artery and 228 normal subjects at a medical center in Taiwan. Information on socio-demographic and smoking status was obtained using a self administered questionnaire. Genotypes of CYP1A1 2A and CYP1A1 2C polymorphisms were determined by polymerase chain reaction or in combination with restriction fragment length polymorphism methods. The results did not show any significant association between CYP1A1 2A polymorphism and CAD risk. However, the CYP1A1 2C G allele was more prevalent in controls (p=0.035) with a dose-response protective effect for CAD. Compared to the A/A genotype, the multivariate logistic regression analysis showed that carrying the CYP1A1 2C G/G genotype was associated with a significantly decreased risk for CAD (OR=0.32, 95% CI=0.15 0.70). The beneficial effect of the CYP1A1 2C G/G genotype was even greater for never smokers than those carrying the A/A genotype (OR=0.23, 95% CI=0.08-0.71). The interaction between genotype and smoking status was not statistically significant. Our findings suggest that the CYP1A1 2C G/G genotype may reduce the risk for CAD in the Taiwanese population and this effect appeared to be more pronounced among never smokers. PMID- 19563928 TI - Gene-environment interactions in chronic pulmonary diseases. AB - The increasing prevalence of asthma and other environmentally induced chronic pulmonary diseases is an important public health concern. It is now generally recognized that the aetiology of these diseases involves a complex interplay between genetic background and exposure to multiple environmental stimuli. This represents a major challenge for the development of new strategies for lung disease prevention and treatment. The completion of the Human Genome Project, the HapMap project, technological advances in genotyping, and the potential of genome wide association analysis has led to a rapid increase in the number of suggested susceptibility genes for asthma and other environmentally induced chronic pulmonary diseases. The genetic studies have used two major methods, i.e., mapping techniques that pinpoint gene loci and studies that identify genes and polymorphisms associated with various asthma mechanisms such as inflammatory mediators. Because the formation of reactive oxygen species is a major aspect of the inflammatory process of the chronic pulmonary diseases, genetic aberrations associated with detoxification enzymes may also shed light on reasons why some people with chronic pulmonary diseases seem more at-risk of exacerbations as a result of environmental insults. This review summarises the present knowledge on the most intriguing potential candidate genes as modifiers of individual susceptibility to these complex diseases. PMID- 19563929 TI - Interaction between Helicobacter pylori, diet, and genetic polymorphisms as related to non-cancer diseases. AB - Helicobacter pylori is a Gram-negative bacterium that infects the stomach of more than half of the world's population. H. pylori infection is an established risk factor for gastric cancer, although it is not sufficient cause for the appearance of cancer, per se. Several studies have investigated the role of this bacterium in non-cancer diseases, including gastritis ulcer, duodenal ulcer, gastroesophageal reflux, cardiovascular diseases, neurodegenerative diseases, ocular diseases, and dermatological disorders. DNA damage and failure in antioxidant defences is a common denominator of many among these pathological conditions. The clinical outcome of H. pylori infection is dependent on many variables, including H. pylori genotype, host health status, host genotype, and host exposure to environmental factors. The role of genetic and environmental factors is reviewed in this paper. Among non-cancer diseases, idiopathic thrombocytopenic purpura appears to show the strongest link with H. pylori. There is an evidence for a role of CagA-positive H. pylori infection in atherosclerosis and ischemic heart disease. On the whole, the major factors playing a pathogenic role in H. pylori-related non-cancer diseases are: (a) host polymorphisms in genes involved in inflammation and protection against oxidative damage, (b) host exposure to dietary genotoxic agents, and (c) bacterial genetic polymorphisms. In conclusion, there is an evidence that mutagenesis-related mechanisms play a pathogenic role in the appearance of non-cancer diseases following H. pylori infection. PMID- 19563930 TI - Transcervical elective superior mediastinal dissection for thyroid carcinoma. AB - OBJECTIVES: To review our results with elective superior mediastinal dissections for thyroid carcinomas. STUDY DESIGN: Retrospective review. METHODS: We searched operative case logs for all patients with thyroid carcinoma treated with an elective superior mediastinal dissection by the senior author (Y.D.) during a 6 year period. Charts were reviewed for demographic information and pathologic results. Elective superior mediastinal dissections were performed when the frozen section was consistent with anaplastic or medullary carcinoma or with a well differentiated carcinoma when there was fixation of the primary tumor to the laryngotracheal complex, there was overt clinically evident paratracheal and/or cervical adenopathy, or the primary tumor measured greater than 2.0 cm in dimension. RESULTS: Thirty-one patients meeting the above criteria were reviewed, and superior mediastinal disease was present in 19 patients (61.3%). Superior mediastinal nodes were positive in 13 (65%) of 20 patients with papillary carcinoma, 0 of 4 with follicular thyroid carcinoma, 4 of 5 patients with medullary thyroid carcinoma, and 2 of 2 patients with anaplastic thyroid carcinoma. Patients with follicular carcinoma had a lower incidence of mediastinal disease (0%) compared with nonfollicular thyroid carcinoma (70.4%), P = .02. Patients with cervical metastasis had an increased incidence of superior mediastinal disease (100% vs 53.3%). CONCLUSIONS: Elective transcervical superior mediastinal dissection was commonly positive in patients with papillary, medullary, and anaplastic thyroid carcinomas. A transcervical approach may be safely performed without sternotomy to the level of the brachiocephalic vein. Further studies are required to determine if performing elective superior mediastinal lymph node dissections will have an impact on survival. PMID- 19563931 TI - The incidence of coexistent autonomic and vestibular dysfunction in patients with postural dizziness. AB - PURPOSE: To evaluate the incidence of coexistent peripheral vestibular dysfunction and cardiovascular autonomic dysfunction in patients undergoing evaluation for dizziness exacerbated by postural changes. MATERIALS AND METHODS: Retrospective case review of 56 sequential patients seen from 2003 to 2006 at a tertiary center for a primary complaint of dizziness who underwent both passive tilt table testing for evaluation of neurocardiogenic etiology and quantitative vestibular testing. The vestibular test battery consisted of alternating bithermal caloric testing; computerized sinusoidal vertical axis rotation (at frequencies 0.01-0.64) with infrared videonystagmography; and oculomotor and positional testing including bilateral Dix-Hallpike, head center supine, and 30 degree supine head turns right and left. RESULTS: Eight of the 56 subjects had caloric weakness. Forty-five subjects (80%) had abnormal tilt table test findings. The incidence of coexistent neurocardiogenic and vestibular test abnormalities was 10.7%. There was no significant association between abnormal tilt table test result and caloric weakness (Fisher exact test; P = .64). The degree of compensation seen on vestibule-ocular reflex gain testing did not affect tilt table findings (chi2; P = .872). CONCLUSIONS: There is no difference in the rate of postural orthostatic intolerance in subjects with evidence of caloric weakness compared with those with normal caloric function. PMID- 19563932 TI - Deep venous thrombosis and pulmonary embolism in otolaryngologic patients. AB - PURPOSE: The objective of this study is to present incidence of deep venous thrombosis and pulmonary embolism in otolaryngologic patients and make recommendations on appropriate thromboprophylaxis. METHODS: The methods used in this study are as follows: 5-year retrospective review in a tertiary care otolaryngology practice and computer-based search from 2000 to 2005. Cases were segregated into ambulatory vs inpatient procedures and malignant vs nonmalignant diagnoses. RESULTS: Six DVTs were identified, 5 with a diagnosed malignancy. One resulted in pulmonary embolism. Inpatient surgeries (1540) and outpatient surgeries (4582) were performed. Eight hundred twenty-four of these were for malignancies (542 inpatient). The incidence of DVT was 0.1%. The incidence in patients with malignancy was 0.6%. CONCLUSIONS: Otolaryngology should be considered a specialty with low thromboembolism risk. Based on this study and recommendations by the American College of Chest Physicians (Northbrook, IL), we recommend early mobilization with the possible adjunct of pneumatic compression stockings in most low-risk otolaryngologic procedures, including most of the outpatient procedures. Moderate-risk patients should be considered for either mechanical or pharmacologic prophylaxis. High-risk patients and patients undergoing high-risk procedures are candidates for pharmacologic thromboprophylaxis. PMID- 19563934 TI - The characteristics of concomitant thyroid nodules cause false-positive ultrasonography results in primary hyperparathyroidism. AB - BACKGROUND: Concomitant thyroid nodules are the most common reason for false positive ultrasonography (US) results in primary hyperparathyroidism. The aims of this prospective clinical study were to evaluate false-positive US results according to the characteristics of concomitant thyroid nodules and to determine which characteristics of thyroid nodules are important. STUDY DESIGN: This prospective study included 120 consecutive patients with primary hyperparathyroidism. The patients were divided into 2 groups according to preoperative US results. Group 1 consisted of 32 patients with false-positive US results and group 2 consisted of 88 patients with true-positive US results. RESULTS: The risk for false-positive US result was increased 25-fold for patients with parathyroid adenoma weight of more than 500 mg (odds ratio [OR], 25; 95% confidence interval [CI], 8.6-74.5), 75-fold for more than 1 posteriorly located thyroid (OR, 75; 95% CI, 19.3-293.4), 358-fold for the presence of exophytic thyroid nodules (OR, 358; 95% CI, 42.3-3036), and 423-fold for the presence of posteriorly located thyroid nodules (OR, 423; 95% CI, 49-3662). CONCLUSION: Although there was no particular characteristic of concomitant thyroid nodules that contributes to false-positive US results, the posteriorly located thyroid nodules were the strongest correlate for the false-positive US results to other features. PMID- 19563933 TI - Extensive paranasal sinus mucoceles: a 15-year review of 82 cases. AB - PURPOSE: The aim of the study was to report the clinical characteristics, management, and outcome of patients with extensive paranasal sinus mucoceles. MATERIALS AND METHODS: In a retrospective study, 82 patients with intracranial or intraorbital extension due to paranasal sinus mucoceles who were surgically treated between 1993 and 2007 were studied. In addition, clinical data, presenting symptoms, clinical features, management, and outcome were analyzed. RESULTS: The study population included 42 males and 40 females, with a mean age of 52.7 years (range, 15-87 years). The most common presenting symptoms were ptosis (27/82, 32.9%) and periorbital swelling (24/82, 29.3%). The main radiologic finding on computed tomography scan was bony defect of lamina papyracea and/or medial superior orbital rim. Sixty-six patients underwent endoscopic sinus surgery; among them, 3 patients had recurrence (3/77, 3.9%) during follow-up periods. CONCLUSION: In our study, a majority of patients with extensive paranasal sinus mucoceles exhibited ophthalmologic symptoms before treatment (81/82, 98.8%). Computed tomography scanning was a feasible tool for preoperative assessment. A satisfactory outcome can be achieved after surgical treatment of endoscopic sinus surgery. PMID- 19563935 TI - Percutaneous feeding tubes in patients with head and neck cancer: rethinking prophylactic placement for patients undergoing chemoradiation. AB - OBJECTIVES: Although intensified therapy has contributed to improved outcomes for patients with head and neck cancer, acute toxicity has increased as well. To lessen the severity of nutritional compromise in these patients, our institutional protocol has been to routinely place feeding tubes before the initiation of therapy. This investigation details the toxicities associated with feeding tube placement and predictors for duration of tube dependence. MATERIALS AND METHODS: The records of the Radiation Oncology Department at Emory Clinic were reviewed for patients receiving definitive radiotherapy between 6/1/2003 and 6/1/2006. The records of the subset of patients with feeding tube placement before the initiation of therapy were then reviewed for toxicities as well as length of time of tube dependence. RESULTS: There were 102 eligible patients. Radiotherapy was delivered with concomitant chemotherapy in all. Median time with feeding tube in place for all patients was 4.4 months (range, 0.2-28.9 months). For 82 patients with eventual tube removal, the median time of tube dependence was 3.8 months (range, 1.4-28.9 months). Risk factors for prolonged tube dependence are analyzed; on multivariate analysis, patient age, T stage, and nodal status remained significant. The most common complication was tube replacement, with 11.8% of all tubes requiring replacement. Infection and pain occurred in 8.8% and 5.9% of patients, respectively. CONCLUSION: Feeding tubes are required for more than 2 months after combined modality treatment of head and neck cancer. They are generally well tolerated, but toxicities are not trivial: more than 10% require replacement and more than 8% of patients develop infection at the insertion site. We are assessing their routine placement in light of these data. PMID- 19563936 TI - Revisiting benign paroxysmal positional vertigo pathophysiology. AB - Benign paroxysmal positional vertigo is the most common peripheral cause of vertigo. Although its pathophysiologic mechanisms remain unclear, different locations have been attributed throughout the last century, from the days of Barany. Disease was initially located by Dix and Hallpike in the utricle, but later, Schuknecht's works elicited the cupulolithiasis and canalolithiasis theories, localizing the pathology to the semicircular canal system and mainly to the posterior one. However, conflicting evidences from temporal bone studies accumulated against this theory, which suggest other explanations. Although this clinical entity is well defined, and can usually be effectively treated with certain physical maneuvers, its pathophysiology is still obscure and is being critically discussed in this article, which reviews the milestones of benign paroxysmal positional vertigo understanding. PMID- 19563937 TI - Cholesteatoma triggering squamous cell carcinoma: case report and literature review of a rare tumor. AB - BACKGROUND: The aim of this study was to report a case of squamous cell carcinoma of the petrous part of the temporal bone associated with a long history of secondary acquired cholesteatoma in a 71-year-old man. PATIENTS AND METHODS: We present the case of a 71-year-old man diagnosed with secondary acquired cholesteatoma in 1950. Treatments consisted of repetitive surgery owing to several relapses. In 2004, he presented with progressive fetid otorrhea. Clinical and computed tomography findings were indicative for relapsing cholesteatoma and a subtotal petrosectomy was performed. RESULTS: Histologic work-up demonstrated a moderately differentiated squamous cell carcinoma. The staging revealed stadium pT3 cN0 cM0. Postoperative treatment consisted of local radiation therapy with intensity-modulated beam geometry with a total of 64.2 Gy in 30 fractions using a simultaneous integrated boost. CONCLUSION: Middle ear carcinoma can arise from acquired cholesteatoma. The pathogenesis of squamous cell carcinoma associated with cholesteatoma has not been elucidated satisfactorily. Due to the complex anatomic features, intensity-modulated radiation therapy is the technique of choice for postoperative radiotherapy. PMID- 19563939 TI - Interdigitating dendritic cell sarcoma of the parotid gland: case report and literature review. AB - Interdigitating dendritic cell sarcoma (IDCS) is an exceedingly rare neoplasm arising from the antigen-presenting cells of the immune system. We report a case of IDCS occurring in a 69-year-old man who presented to an outside institution with a painless mass in his right parotid gland for several months. He presented to our institution after undergoing a superficial parotidectomy. A diagnosis of undifferentiated neoplasm, favoring poorly differentiated carcinoma, was made at that time. He underwent a total parotidectomy and neck dissection at our institution. Microscopically, the tumor was composed of atypical spindle cells involving the parotid gland and an ipsilateral level III lymph node. Immunophenotypic analysis demonstrated positive staining for S100, fascin, vimentin, and HLA-II. Follicular dendritic cell, lymphoid, epithelial, myoepithelial, and melanoma markers were negative. Taken together, the above features were consistent with IDCS. An IDCS of the parotid gland is extremely rare, with only 2 cases reported in the literature. The unusual location and morphological similarity to follicular dendritic sarcoma and other types of soft tissue sarcomas can be a diagnostic challenge. Awareness of this tumor and the use of appropriate markers are crucial in making the diagnosis. The patient did well postoperatively, and he underwent a complete course of postoperative irradiation to the right parotid and neck. PMID- 19563938 TI - A case of increased intracranial pressure after unilateral modified radical neck dissection. AB - OBJECTIVE: This study aimed to study case reports and review the world literature concerning increased intracranial pressure secondary to unilateral radical neck dissection. CASE REPORT: A male patient presented with a painless lump in the left side of his neck. Diagnostic investigation revealed papillary thyroid carcinoma and 4 x 2-cm left cervical lymph node. Total thyroidectomy with modified radical neck dissection on the left side that included removal of the internal jugular vein was undertaken. Eleven days after the operation, the patient presented with a history of headache and diplopia. Clinical examination showed bilateral papilledema and right-sided sixth cranial nerve palsy. A computed tomographic scan and magnetic resonance image of the brain was normal. Subsequent magnetic resonance venography revealed an aplastic contralateral transverse sinus. CONCLUSION: Based on our case, vascular anomalies should be considered in any patient who exhibits signs of increased intracranial pressure after unilateral neck dissection. PMID- 19563940 TI - A case of nasal natural killer/T lymphoma revealed by repeated deep neck abscess. AB - A diagnosis of natural killer (NK)/T-cell lymphoma was clarified after repeated deep neck abscess in a 63-year-old man. The absolute number of lymphocytes was mildly decreased. Lymphopenia induced by latent malignant lymphoma in this case likely caused the immunodeficiency, which induced repeated infection in the head and neck region. We conclude that it is always necessary to consider diseases caused by immunodeficiency conditions, such as malignant lymphoma, into consideration in a case of repeated deep neck abscess. PMID- 19563941 TI - Pyoderma gangrenosum masquerading as necrotizing fasciitis. AB - Necrotizing fasciitis is a fulminant advancing soft tissue infection characterized by widespread fascial necrosis, which can result in significant morbidity and even death. This condition requires prompt diagnosis and aggressive management with extensive surgical debridement and appropriate antibiotic coverage. Pyoderma gangrenosum, in contrast, is a noninfectious inflammatory condition of the skin that typically does not require surgical management. Both conditions can present with extensive ulceration and tissue necrosis, and close clinical-pathologic correlation is required to make the appropriate diagnosis. We present a case in which the diagnosis of pyoderma gangrenosum was initially elusive and serves to illuminate the diagnostic pitfalls in dealing with these entities. PMID- 19563942 TI - Parathyroid carcinoma: clinical presentation and management. AB - Parathyroid carcinoma constitutes a rare cause of hyperparathyroidism. Five patients, presenting with symptoms and clinical signs of parathyroid hyperfunction, were diagnosed with parathyroid carcinoma in our department over a 12-year period. The experience we gained in the management of these tumors is presented. The aim of this study is to elucidate the clinical features of the disease, the surgical treatment, and the sensitivity of the imaging modalities used to enhance the diagnostic ability. Preoperative recognition and intraoperative identification of this rare endocrine malignancy require a high index of clinical suspicion. Intraoperative awareness is needed of the malignant potential of the encountered lesion. PMID- 19563943 TI - A difficult case: sarcoidosis of the middle ear. AB - Sarcoidosis is a common multisystemic granulomatous disorder affecting several organs and tissues. However, the respiratory tract is the region commonly involved in more than 90% of patients, and the middle ear is a direct extension of it. In spite of this, direct middle ear and/or mastoid involvement of sarcoidosis is more rarely seen. Otological involvement may mimic a number of other diseases of the ear; sarcoidosis will probably not be considered prospectively. In addition, pulmonary symptoms of the patients often go unnoticed for some time. We report a patient presenting with hearing loss and tinnitus as the primary manifestation of sarcoidosis of the ear. PMID- 19563945 TI - Fibroma of the tympanic membrane. AB - A primary tumor of the tympanic membrane is very rare. Herein, we describe a patient with a thick tympanic membrane that caused progressive hearing loss. The patient was treated with total resection of the tympanic membrane and was diagnosed with fibroma after histopathologic examination. There was no evidence of recurrence 2 years after the operation, and the patient's hearing was normal. Fibroma may arise in the tympanic membrane and should be regarded as a possible cause of the mass of the tympanic membrane. PMID- 19563944 TI - Leishmaniasis of the auricle mimicking carcinoma. AB - Leishmaniasis of the auricle has been rarely reported in our region of the world, where it is labeled as "Old World Leishmaniasis." It may mimic other pathologies, such as malignancies or other infectious processes. We present a case of an auricular Leishmania lesion which was first suspected to be a carcinoma. Four previous auricular Old World Leishmania cases have been reported. The epidemiology, clinical presentation, diagnosis, and treatment of this entity are briefly reviewed. PMID- 19563946 TI - Fluorodeoxyglucose positron emission tomography/computed tomography in cytologically indeterminate thyroid nodules. PMID- 19563949 TI - Reporting mid- and long-term results of endovascular grafting for abdominal aortic aneurysms using the aortomonoiliac configuration. AB - PURPOSE: This study evaluated the feasibility, efficacy, and durability of a specific aortomonoiliac endograft for the treatment of abdominal aortic aneurysm (AAA) during a midterm follow-up. METHODS: From January 2002 until November 2008, 106 patients (6 women; mean age, 73.37 +/- 7.39 years) were treated for an AAA using an EndoFit aortomonoiliac graft (LeMaitre Vascular, Burlington, Mass). All procedures were elective. RESULTS: Mean follow-up was 34.9 months (SD, 20.08; range, 2-81 months). Long-term data (follow-up >60 months) were available for nine patients, none of which reported any vascular or procedure related complications. Three of the 106 patients (2.83%) died during early follow-up (<30 days); eight died during late follow-up (7.54%). Endograft infection developed in two patients (1.88%), and an aortoduodenal fistula developed in two (1.88%). Also observed were 15 type II (14.15%) and three type I (2.83%) endoleaks. Femorofemoral bypass thrombosis was detected in two patients (1.88%). CONCLUSION: In this retrospective analysis, the aortomonoiliac configuration for elective AAA repair was proven to be safe and efficacious. Midterm and long-term follow-up results in this series compare well with previously reported results for AAA endografting using both bifurcated and aortomonoiliac endoprostheses. PMID- 19563948 TI - Open abdomen treatment following endovascular repair of ruptured abdominal aortic aneurysms. AB - BACKGROUND: Open abdomen treatment (OAT) is considered a lifesaving procedure in patients with abdominal compartment syndrome (ACS) after endovascular or open intervention for ruptured abdominal aortic aneurysms (RAAA). Standardized treatment methods and algorithms for its use are still lacking. The high, published mortality rates may reflect difficulties in detecting and treating ACS, especially in patients treated by emergency endovascular aneurysm repair (eEVAR). Presented are standardized algorithms for OAT, including a new technique using the vacuum-assisted closure (VAC) system developed during 10 years of experience with eEVAR for RAAA. METHODS: We retrospectively analyzed 102 patients with RAAA treated by eEVAR from January 1998 to April 2008. Abdominal decompression was done when intravesical pressure >20 mm Hg or when abdominal perfusion pressure was <50 to 60 mm Hg and concomitant organ deterioration occurred. OAT was initially done with a subcutaneously sutured plastic bag or with a nonsutured zipper drape combined with a VAC device (VAC/ETHIZIP; KCI International Inc, Amstelveen, The Netherlands; Ethicon, Somerville, NJ). All patients were switched to VAC/ETHIZIP as soon as possible. Dressings were generally changed every 3 to 5 days. Intra-abdominal pressure was monitored until stability was observed after delayed direct abdominal closure. RESULTS: Overall 30-day mortality for eEVAR was 13% (13 of 102); 8% (7 of 82) for patients without ACS and 30% (6 of 20) for those with ACS. Decompression for ACS was needed in 20 patients (20%) primarily during the intervention (n = 14) or secondarily in the intensive care unit (n = 6). Six of 20 (30%) patients requiring OAT died 3.0 liters (OR 9.5). Cumulative primary patency at 72 months was 81%; secondary/assisted primary patency was 91%. Limb salvage at 72 months was 89%. Five-year survival was 52%. CONCLUSIONS: These results compare favorably with other methods of treating AGI, especially in patients with multilevel occlusive disease. Principle advantages include acceptable perioperative mortality, low amputation rate, superior durability with excellent long-term patency, and freedom from secondary interventions and recurrent infections. PMID- 19563954 TI - Perioperative diastolic dysfunction during vascular surgery and its association with postoperative outcome. AB - OBJECTIVE: To assess the association of perioperative cardiac dysfunction during elective vascular surgery with postoperative outcome. BACKGROUND: Patients with normal systolic function can have isolated diastolic dysfunction. Routine preoperative evaluation of left ventricular (LV) function does not include an assessment of diastolic function for risk stratification. We hypothesized that perioperative assessment of both diastolic and systolic function with transesophageal echo (TEE) may improve our ability to predict postoperative outcome. METHODS: Perioperative TEE examinations were carried out on patients undergoing elective vascular surgery under general anesthesia. Abnormal systolic function was defined as LV ejection fraction (LVEF) <40%. Left ventricular diastolic function was assessed using transmitral flow propagation velocity (Vp); Vp <45 cm/sec was considered abnormal. We determined the association between LV function and the primary outcome of postoperative adverse outcome, defined as one or more adverse events: myocardial infarction (MI), congestive heart failure (CHF), significant arrhythmia, prolonged intubation, renal failure, and death. RESULTS: Three hundred thirteen patients undergoing vascular surgery were studied. We found that 8% (n = 24) of patients had isolated systolic dysfunction, 43% (n = 134) had isolated diastolic dysfunction, and 24% (n = 75) both systolic and diastolic dysfunction. The most common postoperative adverse outcome was CHF 20% (n = 62). By multivariate logistic regression, we found that patient age, Vp, type of surgery, female gender, and renal failure were predictive of postoperative adverse outcome. CONCLUSION: The presence of perioperative diastolic dysfunction as assessed with Vp is an independent predictor of postoperative CHF and prolonged length of stay after major vascular surgery. Patient age, gender, type of surgery, and renal failure were also predictors of outcome. Perioperative systolic function was not a predictor of postoperative outcome in our patients. PMID- 19563953 TI - The hostile neck does not increase the risk of carotid endarterectomy. AB - INTRODUCTION: Hostile neck anatomy is assumed to be associated with increased surgical risk for patients undergoing carotid endarterectomy (CEA) and is often considered a reason to choose carotid stenting or medical management. This retrospective case-control study evaluated whether, and how much, anatomically hostile necks represent a condition of higher surgical risk of early and late mortality and major or minor morbidity. METHODS: The data for 966 homogeneous CEA patients was prospectively entered in a computer database. Seventy-seven had a hostile neck anatomy due to previous oncologic surgery or neck irradiation, restenoses after CEA, high carotid bifurcation, or bull-like and inextensible neck. A case-control matched-pair cohort study considered sex, age (5-year intervals), and year of operation. Regional anesthesia was used for all operations for atherosclerotic stenosis >or=70%, conforming to the European Carotid Surgery Trial (ECST) in symptomatic and asymptomatic patients, at a single center and by one surgeon or under his direct supervision. RESULTS: The hostile neck patients and the control group were matched for age, sex, carotid related symptoms, degree of stenoses, and main risk factors for cardiovascular diseases. Intraoperative variables were substantially equivalent in the two groups; however, procedure length and clamping time were, respectively, about 22 minutes (P = .0001) and 7 minutes longer (P = .01) in the hostile neck group. Rates of postoperative mortality and neurologic events were equivalent. Peripheral nerve lesions were multiple and significantly more frequent in the hostile neck patients (21% with >or=1 cranial nerve lesion vs 7% of controls, P = .03), yet all were transient and limited to a few months. The subgroups of patients with hostile neck, restenoses, and bull-like inextensible necks required the longest operative and clamping time, and those with bull-like and high bifurcation had the most frequent cranial nerve dysfunctions. At the respective follow-up of 47 and 45 months, survival curves (P = .48) and the incidence of restenoses and fatal and nonfatal strokes were similar (5 and 4, respectively). CONCLUSIONS: Hostile necks led to more complex CEA procedures but without substantial consequences in early and late morbidity and mortality. Most patients with hostile neck can undergo CEA at low risk, with the benefit of effective long lasting stroke prevention similar to standard patients. In our opinion, the more frequent but temporary cranial nerve dysfunctions that occur are not sufficient to consider hostile neck patients noneligible for CEA. PMID- 19563955 TI - Distal vein patch with an arteriovenous fistula: a viable option for the patient without autogenous conduit and severe distal occlusive disease. AB - BACKGROUND: The addition of a distal arteriovenous fistula (DAVF) to improve patency in lower extremity bypass is well described. This report describes a technique of using a distal AVF to enhance a distal vein patch (DVP) in patients without adequate autogenous conduit and who have concomitant severely disadvantaged arterial runoff. METHODS: A retrospective review from May 2002 to May 2007 analyzed 270 tibial bypasses. DVP-AVF was the conduit in 30 bypass grafts. Patient demographics included 16 men, 14 women, diabetes mellitus (67%), and chronic renal failure (20%). All patients had limb-threatening ischemia manifest as rest pain or tissue loss, with 20 patients referred after failed prior revascularization: 11 failed bypasses, and nine failed endovascular interventions. In each case, the only outflow artery available was an isolated tibial segment or a diseased pedal vessel not ordinarily deemed suitable for bypass. At surgery, a common ostium AVF was created between the outflow tibial artery and corresponding tibial vein before DVP construction. Follow-up was 1 to 24 months, with graft function evaluated by pulse examination and duplex surveillance. Primary patency and limb salvage +/- SE were determined by life table analysis using Rutherford criteria. RESULTS: The proximal anastomosis originated from the external iliac (23%), common femoral artery (43%), and superficial femoral artery (33%). Outflow arteries included the anterior tibial (40%), posterior tibial (30%), and peroneal (30%). Venous hypertension in the bypassed limb was noted, but not considered problematic in any patient. Perioperative graft failure occurred in one patient. Six graft failures led to six major amputations (1 above knee; 5 below knee). One patent graft was excised due to infection. Primary patency at 6, 12, 18, and 24 months was 78.3% +/- 6.8%, 78.3% +/- 10.5%, 62.6% +/- 11.1%, and 62.6% +/- 15.6%; limb salvage was 78.7% +/- 6.7%, 78.7% +/- 10.1%, 78.7% +/- 10.1%, and 57.7% +/- 12.5%. CONCLUSION: This early experience describes a modification of the DVP technique in patients with threatened limb loss and severely disadvantaged tibial runoff. The addition of an AVF may reduce outflow resistance, thereby contributing to higher flow rates and improved graft patency. Further investigation is warranted because the DVP-AVF technique may result in acceptable graft patency and limb salvage for patients with no other alternative than amputation. PMID- 19563956 TI - The walking impairment questionnaire: an effective tool to assess the effect of treatment in patients with intermittent claudication. AB - OBJECTIVE: Assessment of walking distance by treadmill testing is the most commonly used method to evaluate the effect of treatment in patients with peripheral arterial disease. However, treadmill testing is time consuming, relatively expensive, and does not adequately reflect real life functional ability. We hypothesized that the Walking Impairment Questionnaire (WIQ) could be an alternative tool to assess objective improvement in functional walking ability of patients with intermittent claudication. METHODS: This was a validation study. It was conducted through the outpatient clinic for vascular surgery. Patients with intermittent claudication were referred for supervised exercise therapy. Treadmill testing (absolute claudication distance [ACD]), WIQ, and quality of life questionnaires (RAND-36 and EuroQol) were administered at study onset and after 3 months of supervised exercise therapy. Responsiveness was determined by mean changes in and correlation coefficients of WIQ, ACD, and quality of life questionnaires. Patients were categorized into quartiles based on the increase in ACD, which were subsequently related to change in WIQ and quality of life. RESULTS: The mean pre- and post-treatment total WIQ scores of 91 patients were 0.45 (0.22) and 0.58 (0.22), respectively. The correlation coefficient between the change in total WIQ score and ACD was 0.331 (P = .004). A 0.1 change in total WIQ score corresponded to a change of 345 meters in ACD. Analysis of the four quartiles compared to an increase in ACD showed that a greater increase in ACD corresponded with a greater increase in WIQ score, from 0.06 to 0.25 (P = .011). CONCLUSION: These data indicate that the WIQ is a valid tool to detect improvement or deterioration in the daily walking ability of patients with intermittent claudication. Hence, the WIQ can be used as an alternative to treadmill testing for objective assessment of functional walking ability, both in daily practice and in clinical trials. PMID- 19563957 TI - Plasma adiponectin as a predictive factor of survival after a bypass operation for peripheral arterial disease. AB - OBJECTIVE: We investigated an association between adiponectin and long-term survival in patients requiring an arterial bypass operation for peripheral arterial disease. METHODS: An enzyme-linked immunosorbent assay kit was used to measure plasma adiponectin levels in 49 patients (38 men, 11 women) before they underwent an arterial bypass operation. Median patient age was 70 years (range, 49-90 years). The study excluded patients with hemodialysis requirement, heart failure, malignant neoplasm, or collagen disease. The symptoms at the first visit were severe intermittent claudication in 27 patients (55%) and critical limb ischemia with rest pain or ulcer, or both, in 22 (45%). RESULTS: Plasma adiponectin levels were a mean 7.8 +/- 5.3 microg/mL (range, 1.0-25.2 microg/mL). Multiple regression analyses revealed that plasma adiponectin was positively correlated with age (r = 0.49, P = .0003) and negatively correlated with body mass index (r = -0.51, P = .0002) and systolic blood pressure (r = -0.41, P = .0059). The Cox proportional hazards model revealed that plasma adiponectin (hazard ratio, 1.30; P = .03) and critical limb ischemia (hazard ratio, 16.67; P = .047) were significant independent predictors of patient survival after a bypass operation. CONCLUSION: Plasma adiponectin could be indicative of mortality after a bypass operation for patients with advanced peripheral arterial disease. PMID- 19563958 TI - Complement activation and plasma levels of C4b-binding protein in critical limb ischemia patients. AB - OBJECTIVE: Critical limb ischemia (CLI) is a peripheral arterial disease manifested by drastically diminished blood flow to the legs, pain at rest, nonhealing wounds, and gangrene caused by atherosclerosis. Significant tissue necrosis is associated with late stage CLI and the patients have a poor prognosis. Necrotic and apoptotic cells activate complement and bind complement inhibitor C4b-binding protein (C4BP). The major isoform of C4BP is composed of seven identical alpha-chains and one beta-chain, here termed C4BP(beta), whereas upon inflammation a normally less abundant isoform is upregulated that is exclusively composed of alpha-chains. Measuring the alpha-chains of C4BP includes both isoforms and is termed total C4BP (C4BP(tot)). The hypothesis of this study was that levels of complement activation and C4BP are predictive for the severity of the disease and that their measurement might be of clinical advantage. METHODS: This was a prospective, single-center study of 259 consecutive patients with CLI admitted to a secondary referral center for vascular diseases. Interventions included evaluation of soluble terminal complement complexes (C5b 9), C4BP(tot) and C4BP(beta), lipid levels, the inflammatory mediators tumor necrosis factor-alpha, interleukin-6, 8-iso-prostaglandin F(2alpha), high sensitivity C-reactive protein, neopterin, plasma homocysteine, and plasma endothelin-1 in plasma as well as resistance to activated protein C and ankle blood pressure. All data were compared with an age-matched population based control group of 219 currently healthy individuals. RESULTS: The data are presented as mean +/- SEM/median. CLI patients showed systemic complement activation (1.17 +/- 0.06/1.13 AU/mL vs 0.69 +/- 0.07/0.59 AU/mL in healthy controls, P < .0001), which was even higher in patients with gangrene (1.33 +/- 0.11/1.28 AU/mL vs 1.1 +/- 0.08/1.0 AU/mL, P = .0264), who also showed increased C4BP levels (421 +/- 28.6/386 microg/mL vs 341 +/- 10.8/318 microg/mL for C4BP(tot), P = .0248; 374 +/- 25.4/332 microg/mL vs 305 +/- 9.5/285 microg/mL for C4BP(beta), P = .0581). C4BP plasma levels were significantly elevated in CLI patients in comparison to healthy controls (351 +/- 8.1/322 microg/mL vs 297 +/- 8.0/288 microg/mL for C4BP(tot), P = .0001; 314 +/- 7.0/287 microg/mL vs 265 +/- 7.0/263 microg/mL for C4BP(beta), P = .0004) and correlated to levels of interleukin-6 (P(tot/beta) = .0048/.0019), high-sensitivity C-reactive protein (P < .0001), leukocyte (P(tot/beta) = .0086/.0043) and platelet count (P = .0001), LDL/HDL ratio (P(tot) = .0151) and HDL (P(tot/beta) = .0047/.0177), but not to tumor necrosis factor-alpha. CONCLUSIONS: Increased complement activation and C4BP plasma levels are related to the degree of tissue necrosis and disease severity of critical limb ischemia. This knowledge in combination with the found correlations to other biomarkers is useful for understanding the pathophysiology of the disease. PMID- 19563959 TI - Midterm results of the surgical treatment of varices by phlebectomy with conservation of a refluxing saphenous vein. AB - BACKGROUND: A new physiopathologic concept within superficial venous insufficiency (SVI) describes ascending progression from the collaterals to the saphenous veins (SV), leading to a treatment that aims to remove the varicose reservoir and not the SVs. This study reports the midterm results of this therapeutic approach. METHODS: This is a retrospective study of patients treated for varices by phlebectomy with conservation of a refluxing SV before July 2004. We evaluated the varicose reservoir by determining the number of zones to be treated (NZT); each lower limb was divided into 32 zones in the preoperative mapping. We performed a clinical and duplex ultrasound examination after 6 months and 1 year, and then once a year until year 4. RESULTS: Amongst 811 lower limbs operated on for first-time varicose veins, 303 in 221 patients (55 men; 166 women), mean age, 52.7 years (range, 20-93 years), were treated by phlebectomy, with conservation of a refluxing SV. All lower limbs operated on presented preoperative SV reflux >0.5 seconds: great SV (GSV), 85.8%; small SV (SSV), 11.9%; and GSV and SSV, 2.3%. The average NZT was 6.05 (range, 2-10). SV reflux was reduced to < 0.5 seconds in 69.6%, 69.2%, 68.7%, 68.0%, and 66.3% of limbs, respectively, after 6 months, 1, 2, 3, and 4 years of follow-up. Symptoms improved or disappeared in 84.2%, 84.2%, 83.4%, 81.4%, and 78.0% of limbs at each annual check-up until year 4. Freedom of varices recurrence was 95.5%, 94.6%, 91.5%, and 88.5%, respectively at 1, 2, 3, and 4 years. When the NZT was >7, the postoperative varicose recurrence was more frequent (odds ratio, 6.82; P = .0001), and the postoperative elimination of SV reflux was more frequent (odds ratio, 4; P = .037) as was symptoms improvement (odds ratio, 2.91; P = .004). When an ostiotruncal SV reflux extended to the malleolus preoperatively, the elimination of the SV reflux was less frequent (47.6% vs 70.3%; P < .05). CONCLUSION: Ablation of the varicose reservoir with conservation of a refluxing SV can be an effective treatment in the midterm for the signs and symptoms of SVI and leads to nonsignificant SV reflux in more than two of three cases. The extent of the varicose reservoir ablation is the key factor determining the hemodynamic and clinical efficacy of this more limited surgical approach. PMID- 19563960 TI - Laparoscopic management of celiac artery compression syndrome. AB - BACKGROUND: Celiac artery compression syndrome (CACS) remains a controversial diagnosis, despite several reported series documenting therapeutic efficacy of CA decompression. Traditional therapy consists of open surgical decompression, but since 2000, five isolated case reports have been published in which CACS has been successfully treated with laparoscopic techniques. This approach was adopted as the sole initial therapy for CACS at the Johns Hopkins Hospital in 2002. This article reports the results of a unique surgical series that triples the reported worldwide experience with this therapy. METHODS: Fifteen patients (median age, 40.6 years) diagnosed with CACS underwent laparoscopic decompression by a single vascular surgeon. CACS was diagnosed by digital subtraction angiography in 14 patients and computed tomography (CT) angiography in one patient, with images acquired in both expiratory and inspiratory phases of respiration. CA decompression was offered after the results of a thorough workup for other pathology were negative, including upper and lower endoscopy, CT scanning, gastric and gallbladder emptying studies, upper gastrointestinal series, and small-bowel follow-through studies. Indications in all patients were abdominal pain and weight loss (average, 9 lbs). The procedure consisted of laparoscopic division of the median arcuate ligament and complete lysis of the CA from its origin on the aorta to its trifurcation. RESULTS: Between November 2002 and September 2007, 15 consecutive patients underwent laparoscopic CA decompression. Median length of follow-up was 44.2 months. There were no operative deaths. Four patients were converted intraoperatively to an open decompression, all for intraoperative bleeding; only one required a blood transfusion. Average operating time was 189 minutes, and the average length of stay was 3.5 days. CA intervention was required in six patients, including three intraoperative procedures (1 patch angioplasty, 1 celiac bypass, 1 percutaneous angioplasty) and six late procedures (2 percutaneous angioplasties, 3 percutaneous stents, 1 celiac bypass). One complication occurred, a severe case of pancreatitis that developed 1 week after discharge. On follow-up, 14 of 15 patients subjectively reported significant improvement, and one patient remains symptomatic with no diagnosis. CONCLUSION: Laparoscopic decompression of the CA may be a useful therapy for CACS, but there is potential for vascular injury, and adjunctive CA intervention is often required. Surgeons should consider laparoscopic CA decompression as a therapeutic alternative for CACS and should participate in the care of patients with this diagnosis. PMID- 19563961 TI - Celiac artery compression syndrome managed by laparoscopy. AB - OBJECTIVE: Celiac artery compression syndrome (CACS) is an unusual condition caused by abnormally low insertion of the median fibrous arcuate ligament and muscular diaphragmatic fiber resulting in luminal narrowing of the celiac trunk. Surgical treatment is the release of the extrinsic compression by division of the median arcuate ligament overlying the celiac axis and skeletonization of the aorta and celiac trunk. The laparoscopic approach has been recently reported for single cases. Percutaneous transluminal angioplasty (PTA) and stenting of the CA alone, before or after the surgical relief of external compression to the celiac axis, has also been used. We report our 7-year experience with the laparoscopic management of CACS caused by the median arcuate ligament. METHODS: Between July 2001 and May 2008, 16 patients (5 men; mean age, 52 years) were treated. Diagnosis was made by duplex ultrasound scan and angiogram (computed tomography [CT] or magnetic resonance). The mean body mass index of the patients was 21.2 kg/m(2). One patient underwent laparoscopic surgery after failure of PTA and stenting of the CA, and two patients after a stenting attempt failed. RESULTS: All procedural steps were laparoscopically completed, and the celiac trunk was skeletonized. The laparoscopic procedures lasted a mean of 90 minutes. Two cases were converted to open surgery for bleeding at the end of the operation when high energies were used. The postoperative course was uneventful. Mean postoperative hospital stay was 3 days. On follow-up, 14 patients remained asymptomatic, with postoperative CT angiogram showing no residual stenosis of the celiac trunk. One patient had restenosis and underwent aortoceliac artery bypass grafting after 3 months. Another patient had PTA and stenting 2 months after laparoscopic operation. All patients reported complete resolution of symptoms at a mean follow up of 28.3 months. CONCLUSIONS: The laparoscopic approach to CACS appears to be feasible, safe, and successful, if performed by experienced laparoscopic surgeons. PTA and stenting resulted in a valid complementary procedure only when performed after the release of the extrinsic compression on the CA. Additional patients with longer follow-up are needed. PMID- 19563962 TI - Retroperitoneal endoscopic release in the management of celiac artery compression syndrome. AB - INTRODUCTION: Celiac artery compression syndrome (CACS) can be treated successfully by division of the median arcuate ligament and celiac plexus fibers. The standard technique is the open approach by an upper midline or left subcostal incision. Only six single cases in which a laparoscopic transabdominal approach for CACS was used have been reported. We prospectively evaluated the feasibility of the endoscopic retroperitoneal approach for treatment of CACS. METHODS: All patients with symptoms suggestive of CACS were evaluated using splanchnic duplex ultrasound scanning, gastric exercise tonometry (GET), and multiplane selective splanchnic angiography. The criteria for treatment were chronic abdominal symptoms, respiratory-dependent CA stenosis, and abnormal GET result. The release was performed by a retroperitoneal endoscopic approach. Anatomic success of the procedure was confirmed by angiography. RESULTS: The endoscopic retroperitoneal approach was used to treat 46 patients with CACS. One patient (2%) required conversion to an open procedure due to suprarenal artery bleeding. Release was ended prematurely in one patient due to a pneumothorax resulting in loss of working space. A postoperative pneumothorax developed in two patients, of which one needed treatment. No other complications were observed. Postoperative angiography during inspiration and expiration showed normal vessel anatomy in 36 of 46 patients. Six of 10 patients with persisting intraluminal stenoses were treated endovascularly. Five of these were successful, which brings the primary assisted anatomic patency for the total group to 89% (41 of 46 patients). Three patients are being observed, and endovascular treatment remains an option in case of insufficient improvement. On median follow-up of 20 months (range, 2-42 months) 41 patients were free of symptoms or showed significant improvement. CONCLUSIONS: The endoscopic retroperitoneal approach for the release of the CA in CACS, with additional endovascular treatment of persistent stenosis, is feasible and effective. Short-term results were comparable with the open procedure. PMID- 19563963 TI - Local lentiviral short hairpin RNA silencing of CCR2 inhibits vein graft thickening in hypercholesterolemic apolipoprotein E3-Leiden mice. AB - OBJECTIVE: Inflammatory responses to vascular injury are key events in vein graft disease and accelerated atherosclerosis, which may result in bypass failure. The monocyte chemoattractant protein-1 (MCP-1)/CC-chemokine receptor (CCR)-2 pathway is hypothesized to play a central role. A murine model for vein graft disease was used to study the effect of local application of lentiviral short hairpin RNA (shRNA) targeted against CCR2. METHODS: A venous interposition was placed into the carotid artery of hypercholesterolemic apolipoprotein E3-Leiden (APOE*3 Leiden) mice to induce vein graft thickening with features of accelerated atherosclerosis. To demonstrate the efficacy of the lentiviral shRNA targeting murine CCR2 (shCCR2) in blocking vein graft disease in vivo, lentiviral shCCR2 or a control lentivirus was used to infect the vein graft locally (n = 8). RESULTS: Vascular CCR2 and MCP-1 messenger RNA expression levels were significantly upregulated during lesion progression in the vein graft. Infection of smooth muscle cells (SMCs) with a lentiviral shRNA targeting shCCR2 completely abolished MCP-1-induced SMC migration and inhibited SMC proliferation in vitro (n = 3 per group). Morphometric analysis of sections of grafts showed a significant 38% reduction in vein graft thickening in the shCCR2-treated mice 4 weeks after surgery (control, 0.42 +/- 0.05 mm(2); shCCR2, 0.26 +/- 0.03 mm(2); P = .007). CONCLUSION: Vascular CCR2 contributes to vein graft disease, and local application of shRNA against CCR2 to the vessel wall prevents vein graft thickening in hypercholesterolemic mice, suggesting that local overexpressing of shRNA using organ-targeted lentiviral gene delivery may be a promising therapeutic tool to improve vein graft disease in bypassed patients. CLINICAL RELEVANCE: Vein graft disease is an important clinical issue that results from an inflammatory response. The monocyte chemoattractant protein (MCP)-1/CC-chemokine receptor (CCR)-2 pathway plays a key role in the initiation and development of vein graft disease. This study demonstrates that perivascular overexpression of short hairpin RNA, targeted against CCR2, inhibits vein graft thickening. These data show that organ-targeted gene therapy against CCR2 in the vessel wall could be a promising therapeutic tool to improve vein graft patency in bypassed patients. PMID- 19563964 TI - Dextran sulfate modulates MAP kinase signaling and reduces endothelial injury in a rat aortic clamping model. AB - OBJECTIVE: Mitogen-activated protein kinases (MAPKs), including JNK, p38, and ERK1/2, noticeably influence ischemia/reperfusion injury (IRI). The complement inhibitor dextran sulfate (DXS) associates with damaged endothelium denudated of its heparan sulfate proteoglycan (HSPG) layer. Other glycosaminoglycan analogs are known to influence MAPK signaling. Hypothetically therefore, targeted intravascular cytoprotection by DXS may function in part through influencing MAPK activation to reduce IRI-induced damage of the vasculature. METHODS: IRI of the infrarenal aorta of male Wistar rats was induced by 90 minutes clamping followed by 120 minutes reperfusion. DXS (5 mg/mL) or physiologic saline (NaCl controls) was infused locally into the ischemic aortic segment immediately prior to reperfusion. Ninety minutes ischemia-only and heparinase infusion (maximal damage) experiments, as well as native rat aorta, served as controls. Aortas were excised following termination of the experiments for further analysis. RESULTS: DXS significantly inhibited IRI-induced JNK and ERK1/2 activation (P = .043; P =.005) without influencing the p38 pathway (P =.110). Reduced aortic injury, with significant inhibition of apoptosis (P = .032 for DXS vs NaCl), correlated with decreased nuclear factor kappaB translocation within the aortic wall. DXS treatment clearly reduced C1q, C4b/c, C3b/c, and C9 complement deposition, whilst preserving endothelial cell integrity and reducing reperfusion-induced HSPG shedding. Protection was associated with binding of fluorescein labeled DXS to ischemically damaged tissue. CONCLUSIONS: Local application of DXS into ischemic vasculature immediately prior to reperfusion reduces complement deposition and preserves endothelial integrity, partially through modulating activation of MAPKs and may offer a new approach to tackle IRI in vascular surgical procedures. CLINICAL RELEVANCE: The purpose of the present study was to determine the role of dextran sulfate (DXS), a glycosaminoglycan analog and complement inhibitor, in modulating intracellular MAPK signaling pathways, reducing complement activation and ultimately attenuating ischemia/reperfusion injury (IRI) in a rat aortic clamping model, in part a surrogate model to study the microvasculature. The study shows a role for DXS in ameliorating endothelial injury by reducing IRI mediated damage and intravascular, local inflammation in the affected aortic segment. DXS may be envisaged as an endothelial protectant in vascular injury, such as occurs during vascular surgical procedures. PMID- 19563966 TI - Anatomy of the carotid sinus nerve and surgical implications in carotid sinus syndrome. AB - BACKGROUND: The carotid sinus syndrome (CSS) is characterized by syncope and hypotension due to a hypersensitive carotid sinus located in the carotid bifurcation. Some patients ultimately require surgical sinus denervation, possibly by transection of its afferent nerve (carotid sinus nerve [CSN]). The aim of this study was to investigate the anatomy of the CSN and its branches. METHODS: Twelve human carotid bifurcations were microdissected. Acetylcholinesterase (ACHE) staining was used to identify location, side branches, and connections of the CSN. RESULTS: A distinct CSN originating from the glossopharyngeal (IX) nerve was identified in all specimens. A duplicate CSN was incidentally present (2/12). Mean CSN length measured from the hypoglossal (XII) nerve to the carotid sinus was 29 +/- 4 mm (range, 15-50 mm). The CSN was frequently located on anterior portions of the internal carotid artery, either laterally (5/12) or medially (6/12). Separate connections to pharyngeal branches of the vagus (X) nerve (6/12), vagus nerve itself (3/12), sympathetic trunk (2/12), as well as the superior cervical ganglion (2/12) were commonly observed. The CSN always ended in a network of small separate branches innervating both carotid sinus and carotid body. CONCLUSION: Anatomical position of the CSN and its side branches and communications is diverse. From a microanatomical standpoint, CSN transection as a single treatment option for patients with CSS is suboptimal. Surgical denervation at the carotid sinus level is probably more effective in CSS. CLINICAL RELEVANCE: Some patients suffering from CSS ultimately require surgical carotid sinus denervation, possibly by transection of its afferent nerve (CSN). This study was performed to investigate the anatomy of the CSN using a nerve-specific ACHE staining technique. Microdissection demonstrated a great variability of the CSN and its branches. Simple high transection of the CSN may lead to an incomplete sinus denervation in patients with CSS. Surgical denervation at the level of the carotid sinus itself may be more effective in CSS. PMID- 19563965 TI - Evidence from cross-sectional imaging indicates abdominal but not thoracic aortic aneurysms are local manifestations of a systemic dilating diathesis. AB - INTRODUCTION: The pathogenesis of aortic aneurysms remains unclear. There is epidemiologic and histologic evidence showing significant differences in aneurysms of the thoracic and abdominal aorta. Studies suggest that abdominal aortic aneurysms (AAA) may represent a local manifestation of a systemic dilating diathesis. It is not known whether thoracic aortic aneurysms (TAA) also have a systemic etiology. The evidence for a systemic dilating diathesis in AAA disease is reviewed and supplemented with an original morphologic study based on computed tomography (CT) comparing nonaneurysmal controls with patients with AAAs and TAAs. METHODS: CT scans performed between January and November 2008 of 150 consecutive patients were examined. The morphology and dimensions of branches of the aorta in 50 TAA patients and 50 AAA patients were compared with 50 nonaneurysmal controls. Measurements of the aorta, common carotid artery (CCA), and superior mesenteric artery (SMA) were taken along with corresponding patient risk factors. RESULTS: Patients were well matched for age, gender, and comorbidity. Mean (SD) right CCA diameter was 9.3 +/- 1.2 in AAA patients vs 8.1 +/- 1 mm in TAA patients (P < .0001) and 7.9 +/- 0.9 mm in controls. Mean left CCA diameter was 9.3 +/- 1.2 mm in AAA patients vs 8.1+/- 0.8 mm in TAA patients (P < .0001) and 7.9 +/- 0.8 mm in controls. There was no significant difference in SMA morphology among the three groups: AAA, 8.6 +/- 1.1; TAA, 8.3 +/- .9; and controls, 8.4 +/- 0.9 mm. Multifactorial modeling accounting for risk factors, age, and gender confirmed that the diameter difference between groups retained independent statistical significance. CONCLUSIONS: This study provides further convincing evidence for a systemic dilating diathesis of elastic arteries in AAAs. It also highlights the differing natures of thoracic and abdominal aortic aneurysmal disease. CLINICAL RELEVANCE: A clear understanding of the pathophysiology of aortic aneurysms is essential. Observational studies have raised the suspicion that abdominal aortic aneurysms are a local representation of a systemic disease of the vasculature. Whether aneurysms of the thoracic aorta have a similar systemic tendency is unknown. Validation of this will open broader avenues of research clarifying understanding of the initiating factors of aneurysms and whether all aneurysms distributed through the arterial tree share the same etiology. This may in turn identify systemic treatments that eventually may allow targeted pharmaceutical management of small aortic aneurysms. PMID- 19563967 TI - Postoperative outcomes in intimal aortic angiosarcoma: a case report and review of the literature. AB - Intimal angiosarcoma is a most unexpected cause of aortic occlusion. We present the case of a 74-year-old woman with intimal angiosarcoma that manifested with the triad of congestive heart failure, acute renal failure, and abdominal angina. A review of the literature and discussion of postoperative outcomes follows. PMID- 19563968 TI - Intravascular mucormycosis as a cause of arm ischemia in an immunocompromised patient. AB - Mucormycosis is an opportunistic fungal infection primarily affecting patients with immunosuppression. We present a case of upper limb ischemia secondary to mucormycosis. A 59-year-old man with myelodysplasia presented with an acutely ischemic arm. The patient underwent numerous revascularisation attempts. Following the final procedure, he developed a cerebral infarct and subsequently died. The most common presentations of mucormycosis are of local character. Once established, it can spread rapidly with vascular invasion and in situ thrombosis. Mortality rate is 76%-96%. In this case, it led to fulminant intravascular coagulopathy resulting in recurrent upper and lower limb ischemia, and finally cerebral infarction. PMID- 19563969 TI - Vascular prosthesis rupture caused by contact with rib stump after thoracic aorta replacement. AB - A 42-year-old male with Marfan syndrome, who had undergone aortic root and total arch replacement for type-A acute aortic dissection at the age of 40, underwent descending aorta replacement with woven Dacron (Vascutek Ltd., Renfrewshire, Scotland) because of pseudoaneurysm at the site of the distal anastomosis and an enlarged pseudolumen of the dissecting descending aorta. The fourth and eighth ribs were cut at their anterior and posterior sites to allow wide exposure of the entire descending aorta. Postoperative computed tomographic scanning showed that the vascular prosthesis posteriorly contacted the eighth rib stump. On the postoperative day 25, the patient collapsed and developed severe hypotension. Emergency thoracotomy revealed a 6 mm in diameter hole on the posterior side of the vascular prosthesis. One day later, the patient died of cardiac dysfunction resulting from sustained hypotension. Electron microscopic examination of the vascular prosthesis showed that the hole was caused by frayed fabric and disrupted polyester fibers. Our experience warns that a woven polyester vascular prosthesis could rupture within 3 weeks of contacting a rib stump. PMID- 19563971 TI - Arteriovenous fistula with multiple anterior tibial artery pseudoaneurysms and peroneal artery rupture following transfemoral embolectomy. PMID- 19563970 TI - Successful surgical treatment of multilevel aortic aneurysms combined with renal transplantation. AB - The concomitant presence of a thoracic aortic aneurysm and an abdominal aortic aneurysm in patients considered for renal transplantation is extremely rare. To our knowledge, this is the first case report of the successful treatment of multilevel aortic aneurysms together with renal transplantation. The treatment modalities in renal transplant patients with concomitant aortic aneurysms are discussed. PMID- 19563973 TI - Surgical infomercials: the ethical price of stardom. PMID- 19563974 TI - Tips and techniques in carotid artery stenting. AB - Significant technical advances have made carotid artery stenting an option for high-risk patients. These advances bring forth new challenges that must be overcome. Preprocedural planning is essential for optimal outcome for every patient given the high risk for significant neurologic complications. In this article we describe a standard approach for performing carotid artery stenting and techniques used to circumvent challenges that may be encountered. In addition, implementation of modifications and advanced techniques in challenging cases may allow successful treatment of carotid stenosis. Maintenance of proficiency in carotid artery stenting requires significant and ongoing experience. PMID- 19563972 TI - Patches for carotid artery endarterectomy: current materials and prospects. AB - Patch angioplasty is commonly performed after carotid endarterectomy. Randomized prospective trials and meta-analyses have documented improved rates of perioperative and long-term stroke prevention as well as reduced rates of restenosis for patches compared with primary closure of the arteriotomy. Although use of vein patches is considered to be the gold standard for patch closure, newer generations of synthetic and biologic materials rival outcomes associated with vein patches. Future bioengineered patches are likely to optimize patch performance, both by achieving minimal stroke risk and long-term rates of restenosis as well as by minimizing the risk of unusual complications of prosthetic patches such as infection and pseudoaneurysm formation. In addition, lessons from bioengineered patches will likely enable construction of bioengineered and tissue-engineered bypass grafts. PMID- 19563975 TI - Five-year results of the getABI study. PMID- 19563976 TI - The ACE trial: a randomized comparison of open versus endovascular repair in good risk patients with abdominal aortic aneurysm. AB - Endovascular repair of infrarenal abdominal aneurysms (EVAR) is currently used in patients with large aneurysm. Two randomized studies, Dutch Randomised Endovascular Aneurysm Management (DREAM) and Comparison of Endovascular Aneurysm Repair with Open Repair in Patients with Abdominal Aortic Aneurysm (EVAR-1), showed favorable early results with EVAR; but at 2 and 4 years, the rates of all cause mortalities were no longer different. Patients in EVAR groups required more reinterventions. These data were confirmed by national audits and large registries. However, there is still uncertainty concerning the durability of the devices, and long-term results are unknown. The ACE (Anevrysme de l'aorte abdominale, Chirurgie versus Endoprothese) trial is a multicenter, prospective randomized trial aimed at assessing the results of EVAR and of open surgery in relatively good-risk patients presenting with an asymptomatic abdominal aortic or aortoiliac aneurysm. The primary end point is death and major complications up to 5 years after randomization. Analysis of results is underway, and publication due by the end of the year. PMID- 19563978 TI - Incorporating outpatient venous procedures into a vascular surgery practice. AB - The use of endovenous procedures to treat superficial venous reflux disease in an exclusively outpatient setting has been growing in popularity during the last decade. Incorporating outpatient venous procedures into an existing vascular surgery practice is a logical step and provides optimal patient care by using the combined expertise and knowledge of devoted vascular surgeons and dedicated supporting staff. This affords the opportunity to treat patients comprehensively and to streamline the evaluation and treatment process for patients with virtually all stages of chronic venous insufficiency. Successful establishment of an outpatient vein care clinic involves making well-informed decisions about workspace, staffing, acquisition of equipment, optimization of reimbursement, and patient recruitment. Separation of the venous practice from the arterial practice both geographically and temporally is critical to success because the clinical and operational needs of these two groups of patients are very different and because the vein practice in many situations may become so robust that it erodes physicians' participation in the arterial side of the vascular practice. In addition to favorable clinical results and increased patient demand, procedures performed on an outpatient basis can be expected to significantly increase revenues. With proper strategy and organization, an outpatient vein practice can round out the existing vascular practice and be a profitable adjunct to an already established vascular surgery practice. PMID- 19563981 TI - There is no sex equality in carotid disease. PMID- 19563982 TI - Regarding "Carotid artery stenting has increased rates of postprocedure stroke, death, and resource utilization than does carotid endarterectomy in the United States, 2005". PMID- 19563986 TI - Foreword. Drug allergy and primum non-nocere. PMID- 19563984 TI - Regarding "C-reactive protein (CRP) elevation in patients with abdominal aortic aneurysm". PMID- 19563987 TI - Preface. Drug hypersensitivity. PMID- 19563988 TI - Approach to the patient with drug allergy. AB - Drug allergies are adverse drug reactions mediated by the specific immune system. Despite characteristic signs (eg, skin rash) that raise awareness for possible drug allergies, they are great imitators of disease and may hide behind unexpected symptoms. No single standardized diagnostic test can confirm the immune-mediated mechanism or identify the causative drug; therefore, immune mediated drug hypersensitivity reactions and their causative drugs must be recognized by the constellation of exposure, timing, and clinical features including the pattern of organ manifestation. Additional allergologic investigations (skin tests, in vitro tests, provocation tests) may provide help in identifying the possible eliciting drug. PMID- 19563989 TI - The pholcodine story. AB - Anaphylactic reactions to neuromuscular blocking agents during general anesthesia constitute a major cause of concern and a great source of debate among anesthesiologists. The authors' recent investigations, taking the striking differences of incidence between Norway and Sweden as the point of departure, have provided valuable insights into the pathogenetic mechanisms and the highly uneven geographical distribution of these rare, but dramatic and notoriously unpredictable, events. Eventually, a cough syrup containing pholcodine emerged as the most likely suspect. This new knowledge led to the withdrawal of the drug from the Norwegian market and to the examination of the role of pholcodine containing drugs in other countries. The present article is a brief summary of the research behind this development. PMID- 19563990 TI - Perioperative anaphylaxis. AB - The incidence of immune-mediated anaphylaxis during anesthesia ranges from 1 in 10,000 to 1 in 20,000. Neuromuscular blocking agents represent the most frequently involved substances, followed by latex and antibiotics, but every drug or substance used may be involved. Diagnosis relies on tryptase measurements at the time of the reaction and skin tests and specific IgE or basophil activation assays. PMID- 19563991 TI - Immediate and delayed reactions to radiocontrast media: is there an allergic mechanism? AB - Radiocontrast media can cause immediate (1 hour) and nonimmediate (>1 hour) hypersensitivity reactions that remain unpredictable and a cause of concern for radiologists and cardiologists. Immediate hypersensitivity reactions resemble anaphylaxis, whereas nonimmediate ones clinically are predominated by exanthemas. Increasing evidence indicates that immediate reactions and nonimmediate skin exanthemas may be allergic reactions involving either contrast media-reactive IgE or T cells, respectively. Skin testing is a useful tool for the diagnosis of contrast media allergy. It may have an important role in the selection of a safe product in previous reactors, although validation data are still lacking. In vitro tests to search for contrast media-specific cell activation are currently under investigation. PMID- 19563992 TI - Heparin allergy: delayed-type non-IgE-mediated allergic hypersensitivity to subcutaneous heparin injection. AB - Itching erythematous or eczematous plaques around injection sites are quite frequent side effects of heparin treatment and clinical symptoms of delayed-type non-IgE-mediated allergic hypersensitivity (DTH) to heparin. For diagnosis, intradermal, patch, and subcutaneous challenge tests with heparins are suitable. In most cases, changing the subcutaneous therapy from unfractionated to low molecular weight heparin or treatment with heparinoids does not provide improvement because of extensive cross-reactivity. Hirudin polypeptides, which exhibit a different chemical structure, are a safe therapeutic alternative for subcutaneous application, however. Importantly, despite DTH to subcutaneously injected heparins, most patients tolerate heparin intravenously. Moreover, in case of therapeutic necessity and DTH to heparins, the simple shift from subcutaneous to intravenous heparin administration without prior testing may be justified. PMID- 19563993 TI - The variable clinical picture of drug-induced hypersensitivity syndrome/drug rash with eosinophilia and systemic symptoms in relation to the eliciting drug. AB - Drug-induced hypersensitivity syndrome (DIHS)/drug rash with eosinophilia and systemic symptoms (DRESS) is a life-threatening adverse reaction characterized by skin rashes, fever, leukocytosis with eosinophilia or atypical lymphocytosis, lymph node enlargement, and liver or renal dysfunction. The syndrome develops 2 to 6 weeks after initiation of administration of a specific drug. It has been demonstrated that various herpesvirus reactivations, in addition to human herpesvirus 6, contribute to internal organ involvement and the relapse of symptoms observed long after discontinuation of the causative drugs. A better understanding of the interplay in the development of DIHS/DRESS has implications for safer and more efficient treatment of this syndrome. PMID- 19563994 TI - Skin testing for IgE-mediated drug allergy. AB - Skin tests with drugs help determine the cause and mechanism of drug hypersensitivity reactions. The diagnosis of adverse drug reactions is based primarily on history and clinical presentation. In type I, IgE-mediated allergic drug reactions, skin prick test and intradermal testing may provide rapid and supportive evidence for diagnosis or exclusion of IgE-mediated reactions. These tests often are more sensitive than laboratory assays for IgE antibodies to drug allergens, which are available only for a few drugs. Because intradermal skin tests occasionally induce adverse events, they should be performed by experienced personnel in an adequate environment. PMID- 19563995 TI - Skin testing in delayed reactions to drugs. AB - Drug skin tests (eg, patch tests, prick tests with delayed readings, intradermal tests [IDT], especially with delayed readings) are used to investigate cutaneous adverse drug reactions (CADR) in delayed hypersensitivity reactions caused by a particular drug. Their value depends on the clinical features of the CADR and on the drug tested. In maculopapular rash (MPR), drug skin tests are of value, beginning with patch tests, and followed: 1) if negative by prick tests (with delayed readings at 24 hours); and, 2) if the injectable form of the drug is available, with IDT with immediate and delayed readings. This article discusses details of the use of patch tests as they apply to patients with various drug reactions. Drug skin tests are useful to study cross-reactivity between suspected drugs. False positive results can occur. The negative predictive value of drug skin tests is approximately 90%. PMID- 19563996 TI - In vitro tests in drug hypersensitivity diagnosis. AB - The diagnosis of a drug hypersensitivity reaction (DHR) is a challenging task because multiple and complex mechanisms are involved. Better understanding of immunologic pathomechanisms in DHRs and rapid progress in cellular-based in-vitro tests can help to adjust the correct diagnostic strategy to individual patients with different clinical manifestations of drug allergy. Thus, drug hypersensitivity diagnosis needs to rely on a combination of medical history and different in vivo and in vitro tests. In this article, the authors discuss current in vitro techniques, most recent findings, and new promising tools in the diagnosis of T-cell-mediated drug hypersensitivity. PMID- 19563997 TI - The basophil activation test in immediate-type drug allergy. AB - Diagnosis of drug allergy involves first the recognition of sometimes unusual symptoms as drug allergy and, second, the identification of the eliciting drug. This is an often difficult task, as the clinical picture and underlying pathomechanisms are heterogeneous. In clinical routine, physicians frequently have to rely upon a suggestive history and eventual provocation tests, both having their specific limitations. For this reason both in vivo (skin tests) and in vitro tests are investigated intensively as tools to identify the disease eliciting drug. One of the tests evaluated in drug allergy is the basophil activation test (BAT). Basophils with their high-affinity IgE receptors are easily accessible and therefore can be used as indicator cells for IgE-mediated reactions. Upon allergen challenge and cross-linking of membrane-bound IgE antibodies (via Fc-epsilon-RI) basophils up-regulate certain activation markers on their surface such as CD63 and CD203c, as well as intracellular markers (eg, phosphorylated p38MAPK). In BAT, these alterations can be detected rapidly on a single-cell basis by multicolor flow cytometry using specific monoclonal antibodies. Combining this technique with in vitro passive sensitization of donor basophils with patients' serum, one can prove the IgE dependence of a drug reaction. This article summarizes the authors' current experience with the BAT in the diagnostic management of immediate-type drug allergy mediated by drug specific IgE antibodies. PMID- 19563998 TI - Provocation tests in drug hypersensitivity. AB - Provocation tests are regarded as the "gold standard" to establish or exclude the presence of hypersensitivity to a certain drugs because they reproduce not only allergy symptoms but other adverse manifestations, irrespective of their pathomechanism. Provocation testing is potentially harmful and should be considered only after balancing the risk-benefit ratio in the individual patient. The reasons for false-positive and false-negative results are numerous, including loss of sensitization, cofactors not being included in the diagnostic procedure, and the potential induction of tolerance during provocation. When conducted by experienced clinicians in a carefully monitored setting, however, drug provocation testing is a safe method to confirm or exclude drug hypersensitivity. PMID- 19564000 TI - [Risk management units]. PMID- 19563999 TI - Rapid desensitization for hypersensitivity reactions to medications. AB - Drug desensitization is the induction of temporary clinical unresponsiveness to drug antigens to which patients have presented severe hypersensitivity reactions. It is typically achieved by gradual reintroduction of small doses of drug antigens at fixed time intervals, and it is aimed at providing increased safety and protection from side effects, including anaphylaxis. Delivery of full therapeutic doses is achieved during desensitization, allowing patients to receive firstline chemotherapy, antibiotics, or monoclonal antibodies, as well as other drugs such as insulin, aspirin, and iron. Desensitizations are high-risk interventions. Inhibition of cellular activation mechanisms occurs during drug desensitization, allowing for the protective clinical outcomes and lack of side effects in the majority of cases, but the cellular and molecular inhibitory mechanisms are incompletely understood. The indication for desensitization protocols can only be done by trained allergists and immunologists and should be implemented as standard of care because of their high success rates and outcomes demonstrated safety profile. PMID- 19564001 TI - [Strategy for implementing and assessing a health care risk management unit in a primary care area]. AB - OBJECTIVE: To describe the setting up of a clinical risk management unit (CRMU) within primary care management, as well as the aims of the project, its implementation phases and the assessment of the results after one year of experience. METHODS: A safety plan was prepared, based on the European Excellence Model (EFQM), to establish a strategic working framework. The plan included 38 proposed actions, associated with criteria elements and 26 indicators to evaluate the selected criteria. RESULTS: A total of 82% of the anticipated actions were implemented in 2007, which included, actions related to teaching and training (15 activities with 237 trainees), spreading of information associated with patient safety, incident analysis (14) and the introduction of specific safe practices (12). Four of those were considered as "generalisable" safe practices and were spread to the rest of the CRMUs in the Autonomous Region of Madrid. The CRMUs have introduced and monitored three processes related to patient safety, participated in a formal programme on the polymedicated elderly, with good results in cover and quality of the indicators. A primary care team (PCT) from the area took part in the first study carried out in Spain on adverse effects in primary care (APEAS Study). CONCLUSIONS: The CRMU can give impetus to strategic lines of safety. The preparation of a strategy defining specific aims has helped in the introduction of patient safety activities and along with the proposed indicators enables the impact of the intervention to be assessed. PMID- 19564002 TI - [Differences in patient satisfaction by gender in users of hospitals in the Cantabrian Health Service]. AB - OBJECTIVE: To study differences in patient satisfaction by gender, among users of hospitals in the Cantabrian Health Service, and if differences were found, to modify organizational policies of the centers in order to improve the quality of care. METHODS: A cross-sectional study was performed in October 2006, using the SERVQHOS questionnaire by telephone. The database of the questionnaire was merged with CMBD database, from which we extracted the length of stay and surgical data. The Student-Fisher t-test was used for quantitative variables and the chi(2) test for qualitative variables. Adjustment was carried out by linear regression. RESULTS: The patient satisfaction is similar in male and female users of the hospitals in the Cantabrian Health Service. Although men are more satisfied on the information to relatives on the place and times for clinical information (92% vs 85.2%; p<0.01), the handing over of the discharge report (95.5% vs 90.9%; p=0.01), and that the report was the definitive (71.6% vs 64.7%; p=0.02). CONCLUSIONS: The differences we found in patient satisfaction by gender do not justify changes in hospital organization to try and improve the quality of care. PMID- 19564003 TI - [Primary care urgent service. Study of patient perceived quality and satisfaction in the Altamira health (Spain) catchment area]. AB - OBJECTIVE: User satisfaction is useful tool to assess the care process, as it provides information on the perceived quality, and can be included as a measurement in the improvement of quality. The Cantabrian Service of Health created the primary care urgent service (PCUS) in 2005, to provide a better service for emergencies, by the rational use of material and human resources, and to satisfy the needs of the population. After a year of its introduction, the Altamira PCUS developed this study, as a means of analysing the opinions of the health users, and to find out their opinions as regards satisfaction and perceived quality, and to see if the needs for which the service was created are satisfied. METHOD: Descriptive cross-sectional study performed during March, 2006. The subjects were patients seen in the PCUS. The research technique used was a self-administered questionnaire, based on SERVQUAL and SERVQHOS, sent by post; the outcomes were considered to be the satisfaction and confidence in the PCUS, and independent variables the sociodemographic characteristics of the users. RESULTS: The patients (81.8%) were generally satisfied with the PCUS service, and 72.5% had confidence in it as a whole. CONCLUSIONS: The setting up of a PCUS has led to an improvement for patients in the Altamira catchment area. This service meets expectations and provides adequate care, as demonstrated by the opinions of the users in the questionnaire, with a high degree of satisfaction (81.8%). PMID- 19564005 TI - [Strategies for patient participation in continuing improvement of clinical safety]. AB - Strategies for patient participation in quality improvement, as an active part of processes or providing relevant information when asked, have progressed to a great extent for the last few years, influenced by the emphasis on patient focused care as a key dimension for quality and, lately, by the emphasis on patient safety -a dimension for which the patient contribution can not be ignored. However, these strategies have not been fully implemented and used in most quality management systems. This article aims to make it easier to select the appropriate strategies for a given context, by describing them, grouped in three main themes (mobilising patients for patient safety; promoting active participation of patients in the prevention of safety incidents; requesting and using the relevant information for quality improvement that patients can provide), illustrating them with examples, and pointing out some of the obstacles for implementing them. PMID- 19564004 TI - [Appropriateness of ketorolac use in a trauma hospital]. AB - OBJECTIVE: To evaluate the suitability of ketorolac and non-steroidal anti inflamatory drugs (NSAIDs) and other analgesic drugs currently used in the hospital. MATERIAL AND METHOD: We have followed the steps to develop a PDCA cycle (plan, do, check, act) or quality improvement cycle. The quality problem was analysed using an Ishikawa diagram. We defined both qualitative quality indicators, those that measure prescription quality, and quantitative ones (defined daily dose, DDD/100BDs), which measure drug consumption, being the objectives to achieve. The study was conducted in all patients admitted to the hospital and who were admitted to orthopaedic and trauma surgery and plastic surgery departments with unit-dose dispensing systems. The strategy used was to give information to physicians through meetings and documentation. Finally, the results were analysed and compared with the initial objectives. RESULTS: The study was performed on 260 patients in the first study period and 292 in the second. Qualitative indicators: intravenous ketorolac use < or =2 days, increased in 25.5% (p<0.001); in patients > or =65 years old at dose < or =60 mg/day it increased 27.7% (p<0.05). Quantitative indicators: in the second study period, ketorolac use decreased (plastic surgery department: 61.8 DDD/100BDs to 14.8), whereas tramadol, ibuprofen and metamizole increased (plastic surgery department: 0 to 14.1 in tramadol, 8.7 to 48.6 in ibuprofen and 50.1 to 71 in metamizole). CONCLUSIONS: Appropriateness of ketorolac, NSAIDs and tramadol use has been achieved, thus improving patient safety. Strategies have been effective. PMID- 19564006 TI - [Usefulness of cardiological written advice in patients with acute coronary syndrome]. PMID- 19564007 TI - Using a magnet to strike gold. PMID- 19564008 TI - Review of cardiovascular magnetic resonance (CMR) in heart failure. Preface. PMID- 19564009 TI - The CMR examination in heart failure. AB - This article presents a rationale for incorporating cardiovascular MR in the evaluation of patients who have heart failure and is organized in four parts. Part I defines the context of cardiovascular MR within standard clinical evaluation and among the many other modalities available for cardiovascular assessment in patients who have heart failure. Part II describes the techniques most useful in treating heart failure. Part III provides recommended approaches for the standard heart failure cardiovascular MR examination. Part IV concludes with perspectives on the appropriate use and incorporation of cardiovascular MR in the evaluation and management of patients who have heart failure. PMID- 19564010 TI - Serial assessment of ventricular morphology and function. AB - Cardiovascular magnetic resonance (CMR) is an accurate, reproducible and well validated imaging technique for the measurement of left ventricular and right ventricular volumes, function, and mass. In patients who have heart failure, CMR is ideally suited both for the initial assessment of fundamental parameters of cardiac function and longitudinal follow-up. Because of its accuracy, the decision to implement therapeutic measures based on cutoff values for ventricular ejection fraction can be made with confidence. Because the above-mentioned parameters correlate with morbidity and mortality, CMR can be used to estimate the prognosis of an individual patient and to obtain surrogate parameters in clinical trials. The process of ventricular remodeling after cardiac injury and reverse remodeling using medical and interventional therapy can be assessed using relatively small sample sizes, which puts CMR in the forefront of imaging techniques in remodeling research. PMID- 19564012 TI - Myocardial viability and revascularization. AB - Cardiovascular MRI can assess multiple markers of myocardial viability in a single examination. Its accuracy is at least equivalent to, if not superior to, that of other currently available noninvasive imaging techniques, including positron emission tomography. The greater spatial resolution afforded by cardiovascular MRI, especially with the delayed-enhancement MRI (DE-MRI) technique, combined with the breadth and depth of correlative pathologic data, makes cardiovascular MRI a particularly powerful tool for detecting viable and irreversibly damaged myocardium. A wealth of clinical data exist, including data from multicenter efforts, to establish DE-MRI as a new gold standard in myocardial viability assessment. As the high accuracy and broad scope of DE-MRI are recognized, the technique will gain wider clinical use for analysis of dysfunctional myocardium and be integrated into the diagnostic and therapeutic algorithm. PMID- 19564011 TI - Evaluation of ischemic heart disease. AB - Approximately two thirds of patients with heart failure have underlying coronary artery disease. In the setting of ischemic heart disease, cardiovascular magnetic resonance has demonstrated usefulness in two ways: for the detection of coronary artery disease and for the assessment of myocardial viability in consideration for revascularization. This article discusses the use of cardiovascular magnetic resonance for the detection of coronary artery disease. The purpose of this article is to provide readers with a brief overview of each of the cardiovascular magnetic resonance techniques, their relative strengths, and their relative weaknesses. Because adenosine stress cardiovascular magnetic resonance is currently the most widely used clinically, it is the primary focus of this article. PMID- 19564014 TI - Cardiovascular magnetic resonance in the evaluation of hypertrophic and infiltrative cardiomyopathies. AB - There is often considerable phenotypic overlap in hypertrophic and infiltrative cardiomyopathies. This overlap creates difficulties, when using routine imaging modalities, in arriving at a conclusive diagnosis. Cardiovascular magnetic resonance (CMR) can make diagnosis easier and more certain. Used with gadolinium contrast agent for tissue characterization, CMR offers a superior field of view and temporal resolution, enabling clinicians to make more confident assessments of etiology. CMR may also be a useful modality for stratifying risk and monitoring treatment responses over time in patients with hypertrophic or infiltrative cardiomyopathies. This article highlights the role of CMR in the assessment and, if relevant, the risk stratification of hypertrophic and infiltrative cardiomyopathies. PMID- 19564013 TI - Identifying the etiology: a systematic approach using delayed-enhancement cardiovascular magnetic resonance. AB - In patients who have heart failure, treatment and survival are directly related to the cause. Clinically, as a practical first step, patients are classified as having either ischemic or non-ischemic cardiomyopathy, a delineation usually based on the presence or absence of epicardial coronary artery disease. However, this approach does not account for patients with non-ischemic cardiomyopathy who also have coronary artery disease, which may be either incidental or partly contributing to myocardial dysfunction (mixed cardiomyopathy). By allowing direct assessment of the myocardium, delayed-enhancement cardiovascular magnetic resonance (DE-CMR) may aid in addressing these conundrums. This article explores the use of DE-CMR in identifying ischemic and non-ischemic myopathic processes and details a systematic approach to determine the cause of cardiomyopathy. PMID- 19564015 TI - Valvular and hemodynamic assessment with CMR. AB - Cardiovascular magnetic resonance is able to provide a comprehensive assessment of valvular and hemodynamic function, including quantification of valve regurgitation and other flows, and accurate cardiac volumes and mass for assessing the effect on both ventricles. Combined with the ability to image all areas of the heart (including difficult areas, such as the right ventricle and pulmonary veins), it is an ideal technique for investigating patients who have heart failure in whom these areas need to be examined. PMID- 19564016 TI - Magnetic resonance imaging of pericardial disease and intracardiac thrombus. AB - Magnetic resonance is known to be a superior modality for the evaluation of pericardial disease and intracardiac masses because of its unmatched capacity for tissue characterization and high spatial resolution. New real-time sequences complement the standard morphologic imaging of the pericardium with dynamic image acquisitions that also can provide hemodynamic information indicative of constriction. Magnetic resonance also is becoming increasingly recognized as a superior modality for the detection and characterization of intracardiac thrombus. This article reviews the use of magnetic resonance imaging for the evaluation of pericardial disease and the detection of intracardiac thrombus, with particular emphasis on the newer pulse sequences currently available for cardiac imaging. PMID- 19564017 TI - Expanding role of cardiovascular magnetic resonance in left and right ventricular diastolic function. AB - This article focuses on the role of cardiovascular magnetic resonance (CMR) in understanding the physiology of diastolic function and on the future applications of CMR as they relate to diastolic function evaluation. CMR has a demonstrated potential to define diastolic function and quantify its properties, in terms of active and passive stages, and its relaxation and compliance characteristics. CMR is also useful for assessing inflow and myocardial velocities, and untwisting properties of the chamber and myocardium, thus providing insights not fully available in other invasive and noninvasive strategies. CMR, which offers the necessary capabilities to evaluate the complex structure of the right ventricle, can serve in the future as the standard for evaluating diastolic function as it currently does for systolic function. PMID- 19564018 TI - Risk stratification for therapeutic management and prognosis. AB - In coronary artery disease (CAD), cardiac magnetic resonance (CMR) imaging can integrate several types of pulse-sequence examinations (eg, myocardial perfusion, cine wall motion, T2-weighted imaging for myocardial edema, late gadolinium enhancement, and CMR angiography) that can provide anatomic, functional, and physiologic information about the heart in a single imaging session. Because of this ability to interrogate myocardial physiology using different pulse sequence techniques within a single CMR session, this technique has been recognized increasingly in many centers as the test of choice for assessing patients who present with cardiomyopathy of undetermined cause. This article first reviews the current evidence supporting the prognosticating role of CMR in assessing CAD and then discusses CMR applications and prognostication in many non-coronary cardiac conditions. PMID- 19564019 TI - Clinical factors affecting the diagnostic accuracy of assessing dilation and curettage vs frozen section specimens for histologic grade and depth of myometrial invasion in endometrial carcinoma. AB - OBJECTIVE: We sought to investigate clinical factors affecting accuracy of dilation and curettage (D&C) and frozen section diagnosis of endometrial cancer. STUDY DESIGN: Clinical parameters affecting concordance of D&C or frozen section compared with final hysterectomy pathology were analyzed in 218 patients with endometrial cancer. RESULTS: The overall concordance of grade between D&C and final hysterectomy findings was 35.2% (62/176). The following factors increased accuracy of D&C: depth of uterus cavity > or = 9 cm (P = .043), deep (> 50%) myometrial invasion (P = .03), P53 positivity (P = .023), grade 2 (P = .01), and grade 3 (P = .048). When comparing frozen section with final hysterectomy findings, the concordance was 69% (58/84) in tumor grade and 87% (67/77) in myometrial invasion. Postmenopausal bleeding (P = .004) and less resistance index of endometrial lesion blood flow (P = .005) increased efficacy of grade diagnosis by frozen section. CONCLUSION: Discordance with hysterectomy assessment was most common for women with D&C or frozen section diagnoses of low-grade superficial cancers. PMID- 19564020 TI - Power over parity: intimate partner violence and issues of fertility control. AB - OBJECTIVE: The purpose of this study was to examine the association between intimate partner violence (IPV), abortion, parity, and contraception use. STUDY DESIGN: We recruited 1463 women for this written questionnaire study of IPV. Patient demographics, contraceptive history, and reproductive history were obtained in the waiting room from patients presenting for gynecologic care. RESULTS: Seventy percent of those eligible participated. Twenty-one percent reported a history of IPV. Partner unwillingness to use birth control, partner desirous of conception, partner creating difficulty for subject's use of birth control, and subjects expressing inability to afford contraception were all positively associated with report of IPV. Each additional pregnancy was associated with 10% greater odds of IPV (95% confidence interval, 1.03-1.17). CONCLUSION: Contraception is more difficult to navigate for women experiencing IPV. Providers should consider prescribing contraceptive methods for IPV victims that are not partner dependent. PMID- 19564021 TI - Poor outcome of indigent patients with peripartum cardiomyopathy in the United States. AB - OBJECTIVE: Peripartum cardiomyopathy (PPCM) patients from Haiti and South Africa have poor survival and poor left ventricular (LV) function recovery compared with patients from the United States. There are no reported studies of PPCM among the African American population in the United States. We evaluated the prognosis of PPCM in a mostly African American population. STUDY DESIGN: We analyzed the clinical and echocardiographic data of 44 (39 African American) patients with PPCM over an 11 year period (1992-2003). RESULTS: Thirty-nine patients were indigent and 5 had health insurance. During a mean follow-up of 24.0 (range, 0.1 264) months, 7 (15.9%) patients died and LV function returned to normal in 14 (35%). CONCLUSION: LV function recovery and survival rates of PPCM patients observed in our study are similar to those reported from Haiti and South Africa and different from what is generally accepted in the United States. PMID- 19564023 TI - In situ formation of stable gold nanoparticles in polymer inclusion membranes. AB - Gold nanoparticles (Au nps) were synthesized in the matrix of a plasticized anion exchange membrane. The membrane was prepared by solvent casting of the solution containing a liquid anion exchanger trioctylmethylammonium chloride (Aliquat 336), a matrix-forming polymer cellulose triacetate (CTA), and a plasticizer dioctyl phthalate (DOP) dissolved in CH(2)Cl(2). For in situ synthesis of Au nps, the membrane samples were equilibrated with a well-stirred solution containing 0.01 mol L(-1)HAuCl(4). AuCl(4)(-) ions were transferred to membrane matrix as an ion pair with Aliquat-336 by an ion-exchange mechanism. In a second step, AuCl(4)(-) ion-loaded membrane samples were placed in a well-stirred 0.1 mol L( 1) aqueous solution of NaBH(4) for reduction. It was observed that 80% of the anion-exchange sites were readily available for the exchange process after formation of the Au nps. The content of Au nps in the membrane was increased either by increasing the concentration of the Aliquat-336 in membrane or by repeating sequential cycles of loading of AuCl(4)(-) ions followed by reduction with BH(4)(-) in the membrane matrix. TEM images of a cross section of the membrane showed that Au nps were dispersed throughout the matrix of the membrane but excluded from the surface. The size distribution of the nps was found to be dependent on Au content in the membrane. For example, 7- to 16-nm Au nps with average size 10 nm were observed in the membrane after the first cycle of synthesis. On increasing the Au content in the membrane by repeating the cycle of synthesis, the size dispersion of nps broadened from 5 to 20 nm without affecting the average size. The lambda(max) (530 nm) and intensity of the surface plasmon band of Au nps embedded in the matrix of membrane were found to remain unaltered over a testing period of a month in the samples kept in water as well as in air under ambient conditions. This indicated that Au nps were quite stable in the membrane matrix. The experimental information obtained by the radiotracers and energy-dispersive X-ray fluorescence (EDXRF) analyses has been used to understand the process of Au nps formation in the membrane matrix. PMID- 19564022 TI - Long-term mortality outcome of victims of major trauma. AB - INTRODUCTION: Although trauma is associated with major acute morbidity and mortality, its long-term outcome is less well defined. We sought to define the rate of readmission to hospital and long-term mortality of victims of major trauma. PATIENTS AND METHODS: All Calgary Health Region residents presenting to the regional trauma program with an injury severity score (ISS) > or = 12 between April 1, 2002 and March 31, 2006 were included. Readmission to hospital within one-year and deaths occurring on or before September 30, 2008 were identified using regional and provincial databases. RESULTS: A total of 2652 incident major trauma episodes occurred among 2630 residents; the median age was 41.3 [interquartile range (IQR); 23.1-59.2] years, 1,915 (72%) were male, the median ISS was 19 (IQR; 16-25), and 717 (27%) required intensive care unit admission. Among 2350 survivors to hospital discharge, 537 readmissions occurred (median length of stay 4.0; IQR; 1.6-7.5 days) among 386 patients within one-year of the incident trauma episode, and 323 (60%) required surgery. Re-admitted patients were older, had higher ISS, had longer initial admission length of stay, and were less likely to have sports related injuries. Two hundred and fifteen (8%) of 2350 survivors to hospital discharge died during the median study follow-up duration of 1543 (IQR; 1181-1934) days. Case-fatality rates for 28 days, 90 days, and 365 days were 304 (11%), 327 (12%), and 370 (14%), respectively. Among survivors to 28 days (n=2348), 66 (3%) suffered delayed one-year mortality (i.e. death occurred between 28 days and 365 days post-trauma). Age > or = 65 years of age, initial hospitalisation for > or = 28 days, and unintentional falls were independently associated with delayed one-year mortality. CONCLUSIONS: Patients with major trauma are at risk for both acute and delayed adverse outcomes. PMID- 19564024 TI - Effective interactions in polydisperse colloidal suspensions investigated using Ornstein-Zernike integral equations. AB - We present a mean of calculating the effective interactions in polydisperse colloidal suspension from liquid state integral equation theory. The method is based on Lado's expansion of correlation functions in a suitable set of orthogonal polynomials. The outlined approach is subsequently used to investigate the effects of polydispersity on the effective potentials for model systems with attractive and repulsive bare interactions. The dominant effect of polydispersity of the smaller species is to weaken the effective potentials between big colloidal particles. This can be exploited as another way of tuning the interactions in colloidal suspensions to match the desired properties. PMID- 19564025 TI - Role of the co-surfactant nature in soybean w/o microemulsions. AB - The influence of the co-surfactant on physicochemical properties of w/o soybean oil microemulsions (MEs) has been studied. In spite of the similarity in phase diagrams, the MEs display remarkable differences when examined by electrical conductivity, dynamic light scattering (DLS), small angle X-ray scattering (SAXS) and linear voltammetry. When different short-chain alcohols were employed as co surfactants, together with sodium dodecyl sulfate (SDS) as surfactant, the DLS results indicated the systems to be monodisperse. Both the electrical conductivity of the MEs and the hydrodynamic radii of the droplets (R(H)) increased with water content while R(H) diminished as temperature increased, no aggregation or percolation of the droplets being observed. In comparison to w/o MEs prepared with 3-methyl-1-butanol, those prepared with 1-pentanol presented higher electrical conductivity and larger limiting currents at a Pt ultramicroelectrode for oxidation of the water occluded into the particles. Finally, from the electrochemical viewpoint the use of 1-pentanol is recommended, no advantage being gained by using any of the other tested alcohols. PMID- 19564026 TI - Synthesis and thermotropic liquid crystalline properties of zwitterionic gemini surfactants containing a quaternary ammonium and a sulfate group. AB - A homologous series of novel zwitterionic gemini surfactants, N,N-dimethyl-N-[3 (alkyloxy)-2-sulfate-propyl]-alkylammonium (referred to as C(m)EOSAC(n)), were synthesized. One of the hydrophilic groups is a sulfate anion, and the other is an ammonium cation. Their thermotropic liquid crystalline properties were investigated by differential scanning calorimetry (DSC), polarizing optical microscopy (POM) and variable-temperature X-ray diffraction (XRD). The results were compared with those for bis(quaternary ammonium) gemini amphiphiles with counterions and catanionic surfactants without a spacer. The C(m)EOSAC(n)s were shown to display improved thermal stability and to form a stable smectic A liquid crystalline phase over a wide temperature region (63+/-0.5 degrees C), which can be attributed to weakening the interactions between the surfactant headgroups. The XRD data confirmed that the two chains of molecules were arranged in a cis configuration with respect to the extended N-CH(2)-CH-SO(4) skeleton in both the solid lattice and the mesophases. In addition, the DSC results showed that the dissymmetry of the hydrophobic chains had a slight effect on the melting point and the clearing point of the C(m)EOSAC(n) compounds. The present investigation points the way to developing stable liquid crystalline materials using gemini type surfactants. PMID- 19564028 TI - Simultaneous orthotopic liver-kidney transplantation with hepatic arterial reconstitution in rats. AB - Although combined liver-kidney (LK) transplantation has been effectively used in patients with end-stage liver disease and concurrent renal failure, a small animal model has been rarely described because of the technical difficulties. Herein, we have described techniques of a modified simultaneous LK transplantation model in rats. We have modified the techniques to perform 47 combined LK transplants in rats with reconstructed hepatic artery and renal vessels by a microvascular sleeve method and an end-to-end anastomosis technique without cross-clamping the vena cava and abdominal aorta, respectively. An average donor operation time was 34.4+/-6.3 min with cold ischemia times of 49.7+/-6.5 min and 61.3+/-4.1 min for liver and kidney grafts, respectively. The total time for recipient operation was 96.8+/-9.4 min with a 4-wk survival rate of 92.3% (36/39). A cumulative hepatic and renal arterial patency rate reached 90.2% (37/41). Normal grafts function tests were observed within the first week post-transplant, as well as normal histopathology studies of the 2 grafts in wk 4 post-transplant. Our method proves to be practical and may contribute to a wider use of the model in the studies of allograft rejection and tolerance induction during combined LK transplantation. PMID- 19564027 TI - Sulindac prevents carcinogen-induced intrahepatic cholangiocarcinoma formation in vivo. AB - BACKGROUND: Intrahepatic cholangiocarcinoma (ICC) incidence and mortality are increasing in the United States and worldwide. ICC etiologies involve chronic inflammation. We hypothesize that the nonsteroidal anti-inflammatory agent sulindac may prevent ICC by targeting cyclooxygenase-1 and -2 (COX-1, -2) as well as COX-independent pathways. MATERIALS AND METHODS: ICC was induced with the carcinogen N-nitrosobis(2-oxopropyl)amine (BOP) in Syrian golden hamsters. Cholangiocarcinogenesis was accelerated by a choline-deficient diet and administration of DL-ethionine and L-methionine. Hamsters were gavaged twice daily for 10 wk with vehicle or sulindac 25, 50, or 75 mg/kg/dose. Harvested livers underwent gross and histopathological examinations. Tissues were analyzed by immunostaining, Western blot, and enzyme-linked immunosorbent assay (ELISA). RESULTS: ICC incidence and multiplicity were decreased in sulindac treatment groups versus control (P < 0.05). In addition, ICC and nontumor lesion sizes decreased in treatment versus control animals. Proliferative indices (Ki-67 immunostaining) decreased and apoptosis (ApopTag immunostaining) increased in treatment versus control (P < 0.05). No changes in COX-1 and -2 protein levels were detected by Western blot. Furthermore, prostaglandin E(2) (PGE(2)) levels were unchanged in treatment and control serum and liver tissues (P > 0.05), suggesting that the antitumor effects of sulindac are mediated by COX-independent mechanisms. Nuclear p65 (activated NF-kappaB) immunostaining decreased (P < 0.05), and protein levels of the NF-kB inhibitor IkappaB-alpha increased in treatment versus control groups. p65 ELISA of liver extracts confirmed decreased NF-kappaB binding activity in sulindac-treated versus control animals (P < 0.05). CONCLUSION: Sulindac effectively prevents experimental cholangiocarcinogenesis, in part by inhibiting the NF-kappaB pathway. PMID- 19564029 TI - Field evaluation of the protective efficacy of Mycobacterium bovis BCG vaccine against bovine tuberculosis. AB - The protective efficacy of Mycobacterium bovis BCG (1 x 10(6) single dose) was evaluated under field conditions. A total of 140 male Holstein Friesian calves, one to two week-old were selected. Two groups of 70 each were formed, one group was vaccinated and the other was injected with a placebo during their second week of age and followed until 12 months of age. The study considered a positive case of tuberculosis to be an animal that had a positive reaction to the three following tests in a row: tuberculin, IFNgamma PPD-B and IFNgamma ESAT6-CFP10 during the 12 months of exposure. The results showed a 59.4% efficacy (IC95%: 47.64-71.16). The non-vaccinated calves were 2.4 times more at risk of becoming infected (IC95%: 1.07-5.68) compared to vaccinated animals. As a complementary test a PCR test was performed using nasal exudates in some animals from both groups using a Mycobacterium complex detection kit. All the positive PCR reactions (5/44) were found in the non-vaccinated animals. These findings suggest that the use of the BCG vaccine, even though it is not capable of protecting 100%, does prevent TB vaccinated animals from excreting bacilli in their nasal secretions at their first year of age. PMID- 19564031 TI - Enhanced monocyte Fc phagocytosis by a homologue of interleukin-10 encoded by human cytomegalovirus. AB - Human cytomegalovirus (HCMV) expresses several homologues of human interleukin 10 (hIL-10) possessing immunomodulatory properties which may promote viral infection by modulating the function of myeloid cells. We examined the phenotype and phagocytic capability of human monocytes exposed to hIL-10, an HCMV-encoded hIL 10 homologue expressed during the productive phase of infection (cmvIL-10), and a differentially spliced form of cmvIL-10 expressed during latent and productive phases of infection, (LAcmvIL-10). hIL-10 and cmvIL-10 upregulated expression of Fcgamma receptors, stimulated phagocytosis of IgG-opsonised erythrocytes and decreased MHC class II (HLA-DR) expression on purified monocytes within 24 h. In contrast, LAcmvIL-10 decreased HLA-DR expression at later times (48 h and 72 h) but did not increase Fcgamma receptor expression. We conclude that cmvIL-10 promotes differentiation of monocytes towards a pro-phagocytic phenotype and that LAcmvIL-10 does not affect monocytes by the same mechanism as cmvIL-10. The significance of these properties to cytomegalovirus pathogenesis is discussed. PMID- 19564030 TI - Effects of allergic airway disease on mouse adenovirus type 1 respiratory infection. AB - Virus infection may contribute to asthma pathogenesis. In turn, a Th2-polarized pulmonary environment may increase host susceptibility to infection. We used a cockroach antigen (CRA) model of allergic airway disease to test the hypothesis that Th2 cytokine overproduction increases susceptibility to mouse adenovirus type 1 (MAV-1). CRA sensitization led to upregulated lung expression of IL-4 and IL-13, lung cellular inflammation, and exaggerated airway mucus production. Following intranasal MAV-1 infection, lung cellular inflammation was more pronounced in CRA-sensitized mice than in unsensitized mice at 7 days post infection but not at a later time point. CRA sensitization did not significantly suppress lung IFN-gamma expression, and lung IFN-gamma expression was upregulated in both CRA-sensitized mice and unsensitized mice over the course of MAV-1 infection. Despite CRA-induced differences in pulmonary inflammation, MAV-1 viral loads in lung and spleen and MAV-1 gene expression in the lung did not differ between CRA-sensitized and unsensitized mice. Our data therefore suggest that MAV 1 pathogenesis is not affected directly or indirectly by the Th2 polarization associated with allergic airway disease. PMID- 19564032 TI - Evolutionary dynamics of Newcastle disease virus. AB - A comprehensive dataset of NDV genome sequences was evaluated using bioinformatics to characterize the evolutionary forces affecting NDV genomes. Despite evidence of recombination in most genes, only one event in the fusion gene of genotype V viruses produced evolutionarily viable progenies. The codon associated rate of change for the six NDV proteins revealed that the highest rate of change occurred at the fusion protein. All proteins were under strong purifying (negative) selection; the fusion protein displayed the highest number of amino acids under positive selection. Regardless of the phylogenetic grouping or the level of virulence, the cleavage site motif was highly conserved implying that mutations at this site that result in changes of virulence may not be favored. The coding sequence of the fusion gene and the genomes of viruses from wild birds displayed higher yearly rates of change in virulent viruses than in viruses of low virulence, suggesting that an increase in virulence may accelerate the rate of NDV evolution. PMID- 19564033 TI - Simultaneous nitrification and denitrification in a CEM (cation exchange membrane)-bounded two chamber system. AB - A novel process was developed to induce a simultaneous oxidation of ammonia and denitrification in a single system consisting of two chambers separated by a cation exchange membrane. One was an anoxic chamber and the other was an aerobic chamber. The maximum mass flux via the membrane was calculated as 0.83mg NH(4)(+) N/m(2)s in a batch test when the initial concentration of NH(4)(+) was 700 mg N/L. And it was observed that NO(3)(-) and NO(2)(-) moved via the membrane in a reverse direction when NH(4)(+) was transported. When the system was operated in a continuous mode by feeding a wastewater containing glucose and NH(4)(+), it was observed that soluble chemical oxygen demand and NH(4)(+) were simultaneously removed showing 99% and 71 approximately 86% of efficiency, respectively. Denitrification occurred in the anoxic chamber and nitrification was carried out in the aerobic chamber. PMID- 19564034 TI - Influence of residual organic macromolecules produced in biological wastewater treatment processes on removal of pharmaceuticals by NF/RO membranes. AB - Increasing attention has been given to pollution of the water environment by pharmaceutical compounds discharged from wastewater treatment plants. High pressure driven membranes such as a nanofiltration (NF) membrane and a reverse osmosis (RO) membrane are considered to be effective for control of pharmaceuticals in wastewater treatment. In practical applications of NF/RO membranes to municipal wastewater treatment, feed water for the membranes always contains organic macromolecules at concentrations of up to 10mg-TOC/L, which are mainly composed of soluble microbial products (SMPs) produced during biological wastewater treatment such as an activated sludge process. In this study, influence of these organic macromolecules on removal of six pharmaceuticals by NF/RO membranes (UTC-60 and LF10) was investigated. Two types of biological treatment (conventional activated sludge process followed by media filtration (i.e., tertiary treatment) and treatment with a membrane bioreactor (MBR)) were examined as pretreatments for NF/RO membranes in this study. In the filtration tests with wastewater effluents, removal of the pharmaceuticals was higher than that seen with deionized pure water spiked with the pharmaceuticals. The increase was significant in the case of the NF membrane. Both alteration of membrane surface properties due to membrane fouling and association of the pharmaceuticals with organic macromolecules contributed to the increase in removal of pharmaceuticals by the membranes. Characteristics of the organic macromolecules contained in the wastewater effluents differed depending on the type of treatment, implying that removal of pharmaceuticals by NF/RO membranes is influenced by the type of pretreatment employed. PMID- 19564035 TI - Ultrafiltration separation of aquatic natural organic matter: chemical probes for quality assurance. AB - Characterization of molecular size of natural organic matter (NOM) is a valuable tool when assessing its effect on the performance of water treatment systems as well as its geochemical origin. Size fractionation can be accomplished by ultrafiltration (UF). Unfortunately, membrane manufacturing generates a range of pore sizes. Many membrane manufacturers use molecular weight cutoff (MWCO) metric based on a 90% retention of given solute after specified duration of filtration. The objective of this study was to characterize the ability of different commercially available UF membranes to separate different size fractions of NOM. The UF membranes characterized were YM (regenerated cellulose, negatively charged) and PB (polyethersulfone, negatively charged) product lines by Millipore. The probes used to represent the size, shape and charge of NOM were polymers (polyethylene glycols (PEGs), dextrans, polystyrene sulfonates (PSSs)), dyes (bromocresol green, congo red, methyl red, methyl orange) and biological molecules (vitamin B-12 and bacitracin). The results show that MWCO definition does not hold for membranes of 5kDa and 10kDa pore openings using most polymers and dyes. The MWCO definition holds for 1kDa membrane for all tested probes. Under natural water conditions PSSs assume random coil configurations that are nearly identical to Suwannee fulvic acid. The results show that PSS agrees with stated MWCOs. The study demonstrates that ultrafiltration is not a simple mechanical sieving process, but that charges on the membrane and the constituent play a significant role in the rejection process. Effective probe size was increased seven- to fourteen-fold by charge interactions between the negative probes and negatively charged membrane. Uncharged molecules larger than specified MWCOs are able to pass through pores (PEGs), while small charged molecules (dyes) do not pass. For probes with low or neutral charges, shape becomes an important factor, with globular being favored over linear structure. Thus, MWCOs cannot be trusted for purposes of NOM size characterization. The study recommends the use of YM 1K, PB 5K and YM 10kDa membranes for comparative-only NOM size ultrafiltration characterization within the 1-10kDa size range. PMID- 19564036 TI - Kinetics of electrooxidation of landfill leachate in a three-dimensional carbon bed electrochemical reactor. AB - The electrooxidation of high strength leachate from an industrial solid waste landfill site was carried out in a three-dimensional carbon bed electrode reactor (TDR). This paper discusses the kinetics and mechanism of electrooxidation on the basis of time course variation of COD, TOC and TKN (total Kjeldahl nitrogen) from the raw leachate. The batch experiments were run at different applied currents (1 3 A) for a period of 6h. A two-stage pseudo-first order reaction kinetics model was developed based on the initial rapid removal of pollutants (Phase I) followed by slow oxidation kinetics (Phase II). About 60-64% COD was removed within 1h with a rate constant 5.83 x 10(-3) min(-1) in Phase I, which was near 5-7 times greater than that of Phase II (0.81-1.03 x 10(-3)min(-1)). The mineralization efficiency was found to be significant in the range 0.83-0.84. The apparent faradic efficiency and specific energy consumption for COD removal were also estimated. The mechanism of electrooxidation was discussed with the help of adsorption, kinetic and SEM results. PMID- 19564037 TI - Perchlorate in human blood serum and plasma: Relationship to concentrations in saliva. AB - The perchlorate anion (ClO(4)(-),MW=99) is present in food, drinking water, groundwater, and surface waters. Exposure to perchlorate is of concern, due to the ability of the anion to disrupt the function of the thyroid gland, and affect the synthesis of thyroid hormones. In this study, liquid chromatography - tandem mass spectrometry (LC-MS/MS) method has been optimized to analyze for perchlorate in blood sera and plasma samples from 84 US donors. In addition, 15 volunteers provided saliva and serum samples concurrently, to enable assessment of the ratio of perchlorate in these two matrices. Recoveries of perchlorate from fortified blanks and from serum/plasma samples were between 92% and 97%. Replicate analysis of blood-matrix spikes had a relative standard deviation (RSD) of <3%, and the relative percent difference (RPD) of repeat analysis of samples was <4%. Perchlorate concentrations in serum and plasma ranged from below the limit of quantitation (0.05ngmL(-1)) to a maximum of 7.7ngmL(-1). Perchlorate concentrations in serum and plasma were log-normally distributed. The mean and median concentrations of perchlorate in 84 serum and plasma samples were 0.32 and 0.17ngmL(-1), respectively. No significant difference existed in perchlorate concentrations between serum and plasma. Analysis of paired saliva and serum samples showed a significant positive correlation for log-normalized perchlorate concentrations (r(2)=0.60) and perchlorate concentrations themselves (r(2)=0.86). The mean saliva:serum concentration ratio of perchlorate was 14:1 (after exclusion of two pairs of outliers). This is the first report to provide measurement data for perchlorate in blood sera and plasma of populations in the US. PMID- 19564038 TI - The effect of quantum dots on synaptic transmission and plasticity in the hippocampal dentate gyrus area of anesthetized rats. AB - Recently, quantum dots (QDs) have attracted widespread interest in biology and medicine. They are rapidly being used as new tools for both diagnostic and therapeutic purposes. Critical issues for further applications of QDs include the assessment of biocompatibility and biosafety of QDs. Most of previous researches concerning QD cytotoxicity focused on in vitro studies. In the present study, the impairments of acute exposure to well-modified and unmodified QDs (streptavidin CdSe/ZnS and CdSe QDs, respectively) on synaptic transmission and plasticity were examined in adult rat hippocampal dentate gyrus (DG) area in vivo. The input/output (I/O) functions, paired-pulse ratio (PPR), field excitatory postsynaptic potential (fEPSP) and population spike (PS) amplitude were measured. The results showed that PPR and long-term potentiation (LTP) were all significantly decreased in these two types of QD-exposed rats compared to those in control rats. While the I/O functions and the amplitudes of fEPSP slope and PS amplitude of the baseline were significantly increased under QD exposure. These findings suggest that exposure to QDs, no matter whether they are well modified or not, could impair synaptic transmission and plasticity in the rat DG area in vivo and reveal the potential risks of QD applications in biology and medicine, especially in the toxin-susceptible central nervous system (CNS). PMID- 19564039 TI - Synthesis, characterization, and in vivo targeted imaging of amine-functionalized rare-earth up-converting nanophosphors. AB - Rare-earth up-converting nanophosphors (UCNPs) have great potential to become a new generation of biological luminescent labels, but their use has been limited by difficulties in obtaining water-soluble UCNPs bearing appropriate functional groups. To address this problem, we report herein a simple and efficient procedure for the preparation of amine-functionalized UCNPs by a modified hydrothermal microemulsion route assisted with 6-aminohexanoic acid. The amine content of the resultant UCNPs has been determined to be (9.5+/-0.8) x 10(-5) mol/g, which not only confers excellent dispersibility in aqueous solution, but also allows further conjugation with targeted molecules such as folic acid (FA) as a ligand. By means of the laser scanning up-conversion luminescence microscopy (LSUCLM) and the in vivo up-conversion luminescence (UCL) imaging under excitation at the CW infrared laser at 980 nm, FA-coupled UCNPs have been demonstrated to be effective in targeting folate-receptor overexpressing HeLa cells in vitro and HeLa tumor in vivo and ex vivo. These results indicated that our UCNPs could be used as whole-body targeted UCL imaging agents. PMID- 19564040 TI - Silk fibroin/hyaluronan scaffolds for human mesenchymal stem cell culture in tissue engineering. AB - The design of new bioactive scaffolds mimicking the physiologic environment present during tissue formation is an important frontier in biomaterials research. Herein, we evaluated scaffolds prepared from blends of two biopolymers: silk fibroin and hyaluronan. Our rationale was that such blends would allow the combination of silk fibroin's superior mechanical properties with the biological characteristics of hyaluronan. We prepared scaffolds with porous microstructures by freeze-drying aqueous solutions of silk fibroin and hyaluronan and subsequent incubation in methanol to induce water insolubility of silk fibroin. Hyaluronan acted as an efficient porogenic excipient for the silk fibroin scaffolding process, allowing the formation of microporous structures within the scaffolds under mild processing conditions. Mesenchymal stem cells were seeded on silk fibroin/hyaluronan scaffolds and cultured for three weeks. Histology of the constructs after cell culture showed enhanced cellular ingrowth into silk fibroin/hyaluronan scaffolds as compared to plain silk fibroin scaffolds. In the presence of tissue-inductive stimuli, in vitro stem cell culture on silk fibroin/hyaluronan scaffolds resulted in more efficient tissue formation when measured by glycosaminoglycan and type-I and type-III collagen gene expression, as compared to plain silk fibroin scaffolds. In conclusion, our data encourages further exploration of silk fibroin/hyaluronan scaffolds as biomimetic platform for mesenchymal stem cells in tissue engineering. PMID- 19564042 TI - Thermodynamic characterization of dissociation rate variations of human leukocyte antigen and peptide complexes. AB - Stability of minor histocompatibility antigen-MHC molecule complexes is a major requirement for the successful presentation of the antigen to T cell receptors. In this letter we show thermodynamic features of the complexes made of a peptide antigen and its three variants to explain molecular basis of variable stability of the complexes. Our analysis suggests that enthalpy is a major factor in determining the stability of the complexes. We also found that the dissociation of the peptides from the complexes exhibits enthalpy-entropy compensation. Two structural features of the complexes, noncovalent chemical bondings and flexibility of the peptides in the complexes, are in a good agreement with our thermodynamic analysis. We expect thermodynamic investigation of peptide antigen MHC protein complexes will provide valuable information on the stability. PMID- 19564041 TI - Chondrogenic and osteogenic differentiations of human bone marrow-derived mesenchymal stem cells on a nanofibrous scaffold with designed pore network. AB - The utilization of 3D scaffolds and stem cells is a promising approach to solve the problem of bone and cartilage tissue shortage and to construct osteochondral (cartilage/bone composite) tissues. In this study, 3D highly porous nanofibrous (NF) poly(L-lactic acid) (PLLA) scaffolds fabricated using a phase separation technique were seeded with multi-potent human bone marrow-derived mesenchymal stem cells (hMSCs) and the constructs were induced along osteogenic and chondrogenic development routes in vitro. Histological analysis and calcium content quantification showed that NF scaffolds supported in vitro bone differentiation. SEM observation showed an altered shape for cells cultured on an NF matrix compared with those on smooth films. Consistent with the morphological change, the gene expression of early chondrogenic commitment marker Sox-9 was enhanced on the NF matrix. NF scaffolds were then used to support long-term in vitro 3D cartilaginous development. It was found that in the presence of TGF beta1, cartilage tissue developed on PLLA NF scaffolds, with the cartilage specific gene expressed, glycosaminoglycan and type II collagen accumulated, and typical cartilage morphology formed. These findings suggest that NF scaffolds can support both bone and cartilage development and are excellent candidate scaffolds for osteochondral defect repair. PMID- 19564043 TI - Fluorescent analogues of quinoline reveal amine ligand loss from cis and trans platinum(II) complexes in cancer cells. AB - Analogues of cytotoxic cis and trans dichloridoplatinum(II) complexes with one ammonia and one aromatic amine (cis- and trans-[PtCl(2)(aromatic amine)(NH(3))]) were synthesised in which the aromatic group was replaced by the fluorescent ligand 7-azaindole (1). Coordination resulted in almost complete quenching of the fluorescence and the ligand had a effect on the biological activities of the cis and trans isomers similar to that previously reported for aromatic amines as is exemplified by them having similar cytotoxicities (IC(50) 3.6(5) and 6.0(19)microM, respectively). Observation of fluorescence following treatment of the cis complex with cysteine, glutathione, or methionine suggests labilisation and subsequent loss of the putative non-leaving group ligands. No such effect was observed for the trans complex which does not experience trans labilisation. Two photon excitation of cells that had been treated with the complexes gave rise to observable fluorescence, suggesting ligand displacement for both complexes. The fluorescence appears to be localised in the lysosomes or late endosomes. These complexes are excellent models of analogues of cytotoxic cis and trans complexes with aromatic amine ligands and can be used to study the metabolism of the non leaving group positions. PMID- 19564044 TI - Increasing extended-spectrum beta-lactamase production and quinolone resistance among Gram-negative bacilli causing intra-abdominal infections in the Asia/Pacific region: data from the Smart Study 2002-2006. AB - OBJECTIVES: The Study for Monitoring Antimicrobial Resistance Trends (SMART) was intended to reveal the evolving profiles of antimicrobial resistance among Gram negative pathogens causing intra-abdominal infections (IAIs). METHODS: Aerobic and facultatively anaerobic Gram-negative isolates were prospectively collected from patients with IAIs in the Asia/Pacific region between 2002 and 2006. Only clinically significant isolates were included for susceptibility tests by the microdilution method. Phenotypic identification of extended-spectrum beta lactamase (ESBL) production was performed on Escherichia coli, Klebsiella spp. and Enterobacter spp., with the measurements of MICs of cefepime alone and in the presence of clavulanic acid. RESULTS: Of 7239 Gram-negative isolates, Enterobacteriaceae accounted for the majority (81.5%), and the three major species were E. coli (42.5%), Klebsiella pneumoniae (15.7%) and Pseudomonas aeruginosa (9.6%). The susceptibility rates of Enterobacteriaceae isolates to cephalosporins decreased, in parallel with the fact that the prevalence of ESBLs increased from 13% in 2002 to 28% in 2006. Among E. coli isolates, the proportion of isolates producing an ESBL varied greatly between countries, ranging from 38.6% in China to 1.2% in New Zealand, and ciprofloxacin susceptibility rate from 41.6% in China to 95.8% in New Zealand. Carbapenems and amikacin were in vitro active against more than 95% of Enterobacteriaceae isolates. CONCLUSIONS: Antimicrobial resistance was frequently present among Gram-negative isolates from IAIs in the Asia/Pacific region. During the study period, there were increasing resistant rates of E. coli isolates to cephalosporins, but wide variations existed among countries. Regular updates of treatment guidelines for this region are necessary for empiric antimicrobial therapy. PMID- 19564045 TI - A prospective study of the diagnostic utility of sputum Gram stain in pneumonia. AB - INTRODUCTION: Sputum Gram stain and culture have been said to be unreliable indicators of the microbiological diagnosis of bacterial pneumonia. The etiological diagnosis of pneumonia is surrounded by great degree of uncertainty. This uncertainty should be and can be calculated and incorporated in the diagnosis and treatment. STUDY OBJECTIVES: To determine the diagnostic accuracy and diagnostic value of sputum Gram stain in etiological diagnosis and initial selection of antimicrobial therapy of bacterial community acquired pneumonia (CAP). DESIGN-METHOD: Prospective study of 1390 patients with CAP admitted January 2002-June 2008, to our institutions. Of the 1390 patients, 178 (12.8%) fulfilled the criteria for inclusion into this study (good-quality sputa and presence of the same microorganism in blood and sputum cultures which was used as gold standard for assessing the diagnostic accuracy and diagnostic value of sputum Gram stain). RESULTS: The sensitivity of sputum Gram stain was 0.82 for Pneumococcal pneumonia, 0.76 for Staphylococcal pneumonia, 0.79 for Haemophilus influenzae pneumonia and 0.78 for Gram-negative bacilli pneumonia. The specificity of sputum Gram stain was 0.93 for Pneumococcal pneumonia, 0.96 for Staphylococcal pneumonia, 0.96 for H. influenzae pneumonia and 0.95 for Gram negative bacilli pneumonia. The positive likelihood ratio (LR+) was 11.58 for Pneumococcal pneumonia, 19.38 for Staphylococcal pneumonia, 16.84 for H. influenzae pneumonia, 14.26 for Gram-negative bacilli pneumonia. The negative likelihood ratio (LR-) was 0.20 for Pneumococcal pneumonia, 0.25 for Staphylococcal pneumonia, 0.22 for H. influenzae pneumonia, and 0.23 for Gram negative bacilli pneumonia. CONCLUSIONS: Sputum Gram stain is a dependable diagnostic test for the early etiological diagnosis of bacterial CAP that helps in choosing orthological and appropriate initial antimicrobial therapy. PMID- 19564046 TI - Severe affective and behavioral dysregulation in youth is associated with increased serum TSH. AB - BACKGROUND: The relationship of bipolar disorder (BD) and altered thyroid function is increasingly recognized. Recently, a behavioral phenotype of co occurring deviance on the Anxious/Depressed (A/D), Attention Problems (AP), and Aggressive Behavior (AB) syndrome scales has been identified as the Child Behavior Checklist Dysregulation Profile (CBCL-DP), which itself has been linked to BD. This study tested for differences in thyroid function within a sample of n=114 psychiatric children and adolescents with and without the CBCL-DP. METHOD: A CBCL-DP score was generated based on the composite of the crucial CBCL syndrome scales (A/D, AP, AB). Participants with a CBCL-DP score >or=2.5 SDs above average constituted the CBCL-DP subgroup (n=53). Those with CBCL-DP scores of 1 SD or less above average percentile were regarded as controls (n=61). Groups were compared regarding serum levels of TSH, fT3 and fT4. RESULTS: In participants showing the CBCL-DP, basal serum TSH was elevated compared to controls. More CBCL DP subjects than controls showed subclinical hypothyroidism. No differences were observed for serum fT3 and fT4 levels. CONCLUSIONS: This is the first study to demonstrate associations between CBCL-DP and subclinical hypothyroidism. Future research should address the long-term outcome of CBCL-DP with coexisting hypothyroidism, the potential benefits of supplementation with thyroid hormone, and the association between severe dysregulation and the bipolar spectrum. PMID- 19564047 TI - Six year follow-up of a clinical sample of self-harm patients. AB - BACKGROUND: Mortality from suicide and other causes is significantly increased in patients who engage in self-harm, but their long-term morbidity and quality of life are poorly defined. As the majority of self-harm patients are under the age of 35 years, understanding their longer term health outcomes is important if we are to adequately manage their care. The aim of this study was to investigate the long-term mortality, morbidity and quality of life of such patients. METHOD: A representative cohort of patients who had presented to hospital following an episode of self-harm was traced after 6 years. Mortality and repetition of self harm were primary outcome measures. Psychiatric morbidity and indices of quality of life, and social functioning were also obtained. RESULTS: 143/150 (95.3%) patients were traced after a mean of 6.2 years. Eight (5.6%) had died during follow-up, significantly more than general population estimates (p < or = 0.001), four of these (2.8%) by probable suicide. Further self-harm occurred in 58/101 (57.4%) participants; 70/97 (72.2%) fulfilled criteria for at least one psychiatric disorder, and 51.3% screened positive for harmful use of alcohol. Measures of health status (EQ-5D and SF36-II) were significantly lower (p<0.001) than in the general population. LIMITATIONS: Due to the nature of this population group the attrition rate at 6 years is high; although this is the most complete such study to date. CONCLUSION: Despite positive outcomes in some patients, overall levels of mortality, morbidity, and harmful use of alcohol are high, whilst quality of life is reported as low. This has significant implications for the long-term management of this group. PMID- 19564048 TI - The norepinephrine transporter gene modulates the relationship between urban/rural residency and major depressive disorder in a Chinese population. AB - The norepinephrine transporter (NET), which is involved in the neurotransmission of norepinephrine (NE), may play an important role in the development of major depressive disorder (MDD). Recent studies have suggested that a gene-environment interaction may confer susceptibility to depression. The aims of this study were to test modifying effects of the NET gene on the association between residency and MDD, as well as to reveal the relationship between gender and this gene environment interaction in MDD. This study recruited a total of 442 patients with MDD and 393 controls. Residence was defined as reported in the data of the 5th Chinese census. Logistic regression models were used to analyze gene-environment interactions. A gene-environment interaction between the G1287A polymorphism and residency was found in the female sample. In addition, and odds ratio analysis showed that only rural women carrying the G/G genotype of the G1287A polymorphism were susceptible to MDD, but others were not. To our knowledge, this is the first report showing that the G1287A polymorphism modifies rural residency as a risk factor for MDD. These findings support the possibility that the NET gene is an important factor in susceptibility to MDD in a Han Chinese population. PMID- 19564049 TI - Preliminary evidence on the association between XBP1-116C/G polymorphism and response to prophylactic treatment with valproate in bipolar disorders. AB - The 116C/G polymorphism in the promoter region of XBP1 is known to be associated with bipolar disorders. The G allele of the XBP1-116C/G polymorphism has reduced XBP1-dependent transcription activity compared with the C allele. Valproate treatment has been known to rescue the impaired response of cells with the G allele. We investigated the hypothesis that the G allele of XBP1-116C/G has better prophylactic treatment response to valproate compared to the C allele. This study involved 51 patients with bipolar disorder who were treated with valproate for prophylactic treatment. Prophylactic treatment response to valproate was retrospectively assessed using a scale described by Grof et al. [Grof, P., Duffy, A., Cavazzoni, P., Grof, E., Garnham, J., MacDougall, M., O'Donovan, C., Alda, M., 2002. Is response to prophylactic lithium a familial trait? Journal of Clinical Psychiatry 63, 942-947.]. We found that the patients with the G allele of XBP1-116C/G showed a better prophylactic treatment response to valproate compared to the C allele. This result is in agreement with in-vitro data showing that valproate ameliorates the endoplasmic reticulum (ER)-stress response compromised by the G allele. PMID- 19564050 TI - The NimStim set of facial expressions: judgments from untrained research participants. AB - A set of face stimuli called the NimStim Set of Facial Expressions is described. The goal in creating this set was to provide facial expressions that untrained individuals, characteristic of research participants, would recognize. This set is large in number, multiracial, and available to the scientific community online. The results of psychometric evaluations of these stimuli are presented. The results lend empirical support for the validity and reliability of this set of facial expressions as determined by accurate identification of expressions and high intra-participant agreement across two testing sessions, respectively. PMID- 19564051 TI - Association of plasma homocysteine and methylenetetrahydrofolate reductase C677T gene variant with schizophrenia: A Chinese Han population-based case-control study. AB - This study examined the association of plasma homocysteine levels and the methylenetetrahydrofolate reductase (MTHFR) gene polymorphism with schizophrenia in the Han population residing in northern China. We detected the MTHFR C677T genotype in 123 schizophrenia patients and compared it with the genotype of 123 controls by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In addition, by using the cyclophorase method, the plasma homocysteine concentration in 62 schizophrenia patients was determined and then compared with that in 62 controls; these 62 patients and 62 controls were a subset of the 123 patients and 123 controls. We found that the homocysteine levels in the patients were significantly higher than those in the controls. The frequency of homozygosity for the 677T allele of the MTHFR gene was higher in the patient group than in the control group, and the difference between the two groups was statistically significant for both the MTHFR genotype and the frequency of allele homozygosity. A significant difference was observed in the plasma homocysteine levels among the different genotypes in the patient and control groups. In conclusion, both elevated plasma homocysteine levels and variation in the MTHFR 677C-->T gene are related to increased rates of schizophrenia and are risk factors for schizophrenia. PMID- 19564052 TI - Pegylated nanoliposomes remote-loaded with the antioxidant tempamine ameliorate experimental autoimmune encephalomyelitis. AB - Reactive oxygen species are involved in the pathogenesis of multiple sclerosis (MS), Parkinson's disease and neurodegenerative diseases. Here we report that Tempamine (TMN), a stable radical with antioxidant and proapoptotic activities, when encapsulated in the intraliposome aqueous phase of pegylated (<100 nm) nanoliposomes (nSSL), is efficient in inhibiting experimental autoimmune encephalomyelitis (EAE) in mice. The TMN is remote-loaded into nSSL by an intraliposome high/extraliposome low transmembrane ammonium sulfate gradient. Biodistribution studies of nSSL-TMN labeled with the liposome non transferable non metabolizable (3)H-cholesteryl hexadecyl ether show that almost 3% of the injected dose of liposomes reached the brain of the EAE mice, compared with less than 1% in the control healthy mice. This accumulation in the brain, combined with the fact that TMN demonstrates a controlled slow release out of the nSSL, may explain the superior therapeutic activity of nSSL-TMN over free TMN. Our results suggest that the study of nSSL-TMN for therapy of MS, and other neurodegenerative diseases involving oxidative damage, is worth pursuing. PMID- 19564053 TI - Long lasting recruitment of immune cells and altered epi-perineurial thickness in focal nerve inflammation induced by complete Freund's adjuvant. AB - Immune-mediated nerve inflammation is involved in many painful states in humans, and causes axonal and behavioral changes in rats. While models of nerve inflammation have been characterized using electrophysiological and behavioral methods, the presence of immune cells has not been fully assessed. We inflamed rat sciatic nerves using complete Freund's adjuvant and quantified the presence of ED-1 macrophages and TCR-alphabeta T-cells for up to 12 weeks. We report that these immune cells are prominent extraneurally up to 12 weeks following the induction of inflammation. This observation does not easily correlate with inflammation-induced axonal mechanical sensitivity, which peaks within 1 week and is resolved after 8 weeks. PMID- 19564055 TI - On the use of conventional and tissue Doppler echocardiography in patients with beta-Thalassemia major and myocardial iron-overload: preliminary data by a single centre study. PMID- 19564054 TI - Linking genotoxic responses with cytotoxic and behavioural or physiological consequences: differential sensitivity of echinoderms (Asterias rubens) and marine molluscs (Mytilus edulis). AB - Integrated laboratory studies addressed multiple biomarker responses in the sea star (Asterias rubens) and the blue mussel (Mytilus edulis) exposed to a range of concentrations of direct and indirect acting genotoxins: methyl methane sulfonate (MMS) and cyclophosphamide (CP; an environmentally relevant anti-cancer pharmaceutical), respectively, in order to determine if the expressed genotoxicity has knock-on effects at the higher levels of biological organisation. The experimental design aimed to concurrently evaluate biomarkers of behavioural and physiological conditions (i.e. 'righting time' and 'clearance rate' for sea stars and mussels, respectively) in addition to cytotoxicity (neutral red retention assay), induction of micronuclei (Mn) and DNA strand breaks (as determined by the Comet assay). The protocol also included the determination of the maximum tolerated concentration (MTC), prior to genotoxic evaluation. The 3d MTC, as determined by the survival of the organisms, showed sea stars to be more sensitive than mussels to MMS (18 and 32 mg L(-1), respectively) and CP (56 and 180 mg L(-1), respectively). For both species and chemicals, cytotoxicity was not found to be significantly different compared to controls. Apart from the MMS exposure to sea stars (which showed 100% mortality at higher concentrations after 5d exposure), clear dose-response relationships were observed for both genotoxicity endpoints in each species. Following exposure to CP, good correlations were also found between the behavioural and physiological responses and genetic damage in each species (sea stars-MN vs. RT: R=0.73; Comet vs. RT: R=0.91; mussels-MN vs. CR: R=0.69; Comet vs. CR: R=0.72). This integrated approach, applying non-invasive assays to simultaneously determine the responses at different levels of biological organisation, indicates the potential value of behavioural and physiological measures in determining the toxicity of chemicals to marine organisms and highlights also the relevance of including adult echinoderms in environmental studies. PMID- 19564056 TI - Les liaisons dangereuses: Tako-Tsubo syndrome after an adulterous intercourse in an elderly male. AB - We report the case of an 81-year-old man who presented with chest pain after an adulterous intercourse with a young woman. At echocardiography a typical apical ballooning phenomenon was detectable and completely reversed after one week. Coronary angiography did not show significant coronary atherosclerosis. PMID- 19564057 TI - Biomedical applications of distally controlled magnetic nanoparticles. AB - Nano-sized magnetic particles are increasingly being used across a wide spectrum of biomedical fields. Upon functionalization to enable specific binding, magnetic particles and their targets can be conveniently positioned in vitro and in vivo by the distal application of magnetic fields. Furthermore, such particles can be magnetically heated after reaching their in vivo targets, thus inducing localized cell death that has a considerable therapeutic value in, for instance, cancer therapy. In this context, innovative biomedical research has produced novel applications that have exciting clinical potential. Such applications include magnetically enhanced transfection, magnetically assisted gene therapy, magnetically induced hyperthermia and magnetic-force-based tissue engineering, and the principles and utilities of these applications will be discussed here. PMID- 19564058 TI - Applicability of a microbial Time Temperature Indicator (TTI) for monitoring spoilage of modified atmosphere packed minced meat. AB - The applicability of a microbial Time Temperature Indicator (TTI) prototype, based on the growth and metabolic activity of a Lactobacillus sakei strain developed in a previous study, in monitoring quality of modified atmosphere packed (MAP) minced beef was evaluated at conditions simulating the chill chain. At all storage temperatures examined (0, 5, 10, 15 degrees C), the results showed that lactic acid bacteria (LAB) were the dominant bacteria and can be used as a good spoilage index of MAP minced beef. The end of product's shelf life as revealed by the sensory evaluation coincided with a LAB population level of 7 log(10) CFU/g. For all temperatures tested, the growth of L. sakei in the TTI resembled closely the growth of LAB in the meat product, with similar temperature dependence of the micro(max) and thus similar activation energy values calculated as 111.90 and 106.90 kJ/mol, for the two systems, respectively. In addition, the end point of TTI colour change coincided with the time of sensory rejection point of the beef product during its storage under isothermal chilled temperature conditions. The estimated activation energy, E(alpha), values obtained for parameters related to the response of DeltaE (total colour change of the TTI) describing the kinetics of colour change of the TTI during isothermal storage (i.e. the maximum specific rate of DeltaEpsilon evolution curve, micro(DeltaEpsilon), and also the reciprocal of t(i), time at which half of the maximum DeltaEpsilon is reached), were 112.77 and 127.28 kJ/mol, respectively. Finally, the application of the microbial TTI in monitoring the quality deterioration of MAP minced beef due to spoilage was further evaluated under dynamic conditions of storage, using two separate low temperature periodic changing scenarios, resembling the actual conditions occurring in the distribution chill chain. The results showed that the end point of TTI, after storage at those fluctuating temperature conditions, was noted very close to the end of product's sensorial shelf life. This finding points to the applicability of the developed microbial TTI as a valuable tool for monitoring the quality status during distribution and storage of chilled meat products, which are spoiled by lactic acid bacteria or other bacteria exhibiting similar kinetic responses and spoilage potential. PMID- 19564060 TI - Synthesis of novel bioactive derivatives of 3-(4-chlorophenyl)-2-hydrazino 5,6,7,8-tetrahydrobenzo(b)thieno[2,3-d]pyrimidine-4(3H)-ones. AB - A series of triazolo[4,3-a]tetrahydrobenzo(b)thieno[3,2-e]pyrimidine-5(4H)-ones (12a-n) were synthesized and evaluated for CNS depressant, skeletal muscle relaxant and anticonvulsant activities by photoactometer, Rotarod and pentylenetetrazole induced the convulsions method respectively in Swiss albino mice. Diazepam was used as standard drug. The five derivatives 12b, 12c, 12d, 12i and 12m showed the CNS depressant and skeletal muscle relaxant activities comparable to those of diazepam at a dose of 5mg/kg. These derivatives also exhibited good activity when tested for anticonvulsant activity in mice at different dose levels. The ED(50) values for these derivatives are in the range of 4.40-9.33 mg/kg. PMID- 19564059 TI - Age-associated cerebral atrophy in mouse lemur primates. AB - We assessed the regional brain atrophy in mouse lemur primates from 4.7T T2 weighted magnetic resonance images. Thirty animals aged from 1.9 to 11.3 years were imaged. Sixty-one percent of the 23 animals older than 3 years involved in the study displayed an atrophy process. Cross-sectional analysis suggests that the atrophy follows a gradual pathway, starting in the frontal region then involving the temporal and/or the parietal part of the brain and finally the occipital region. Histological evaluation of five animals selected according to various stages of atrophy suggested that extracellular amyloid deposits and tau pathology cannot explain by themselves this atrophy and that intracellular amyloid deposition is more closely linked to this pathology. This study suggests that most of the age-related atrophy occurring in mouse lemurs is caused by one clinical, evolving, pathological process. The ability to follow this pathology non-invasively by MRI will allow to further characterize it and evaluate its relationship with neuropathological lesions that are involved in human diseases such as Alzheimer. PMID- 19564061 TI - The practice assessment of student nurses by people who use mental health services. AB - This paper reports on a participatory action research project focusing upon service-user involvement in the assessment of student mental health nurses in practice. The principle of student nurses being assessed by service-users was found to be desirable in theory. However, its implementation proved to be fraught with difficulty especially with regard to the concerns about increased workload and feelings of vulnerability amongst students. It is recommended that the model of assessment should be adjusted to that of review and that a program of service users reviewing student nurses on placement be implemented and evaluated. PMID- 19564062 TI - The impact of malnutrition and nutrition-related factors on the development and severity of pressure ulcers in older patients receiving home care. AB - BACKGROUND & AIMS: To investigate the impact of nutritional status and nutrition related factors on the development and severity of pressure ulcers acquired in the home care setting. METHODS: Two hundred and seven home care offices in Japan were selected at random and 290 patients with home-acquired pressure ulcers and 456 patients without pressure ulcers were analyzed. Data on nutritional status, caregiver knowledge, and health professional's nutritional management were collected. Pressure ulcers were categorized as superficial or full-thickness. RESULTS: Malnutrition was significantly and most strongly associated with higher rate of the pressure ulcer after adjusting for other risk factors (OR, 2.29; 95% CI, 1.53-3.44). Assessment of the patient's nutritional status and adequate dietary intake by a health professional were significantly associated with lower odds for developing pressure ulcers (OR, 0.43, 0.47; 95% CI, 0.27-0.68, 0.28 0.79, respectively). Malnutrition was also significantly and most strongly associated with more severe pressure ulcers (OR, 1.88; 95% CI, 1.03-3.45). Assessment of a caregiver's nutritional knowledge by a health professional was a significant preventive factor for severe pressure ulcers. CONCLUSION: The quality of home care for risk factors such as pressure redistribution has improved, making nutritional management a more crucial factor in pressure ulcer prevention. PMID- 19564063 TI - Voluntary sway and rapid orthogonal transitions of voluntary sway in young adults, and low and high fall-risk older adults. AB - BACKGROUND: Falls amongst older people have been linked to reduced postural stability and slowed movement responses. The objective of this study was to examine differences in postural stability and the speed of response between young adults, low fall-risk older adults, and high fall-risk older adults during voluntary postural sway movements. METHODS: Twenty-five young adults (25+/-4 years), and 32 low fall-risk (74+/-5 years), and 16 high fall-risk (79+/-7 years) older adults performed voluntary sway and rapid orthogonal transitions of voluntary sway between the anterior-posterior and medial-lateral directions. Measures included reaction and movement time and the amplitudes of the centre of pressure, centre of mass, and the separation distance between the centre of pressure and centre of mass. FINDINGS: Both fall-risk groups compared to the young had slower reaction and movement time, greater centre of pressure and/or centre of mass amplitude in the orthogonal (non-target) direction during voluntary sway, and reduced anterior-posterior and medial-lateral separation between the centre of pressure and centre of mass during voluntary sway and orthogonal transitions. High compared to low fall-risk individuals had slower reaction and movement time, increased non-target centre of mass amplitude during voluntary sway, and reduced medial-lateral centre of pressure and centre of mass separation during voluntary sway and orthogonal transitions. INTERPRETATION: Age related deterioration of postural control resulted in slower reactive responses and reduced control of the direction of body movement during voluntary sway and orthogonal transitions. Slower postural reaction and movement time and reduced medial-lateral control of the centre of mass during voluntary sway movements are associated with increased fall-risk in community-living older people. PMID- 19564064 TI - Geochemical processes of mercury in Wujiangdu and Dongfeng reservoirs, Guizhou, China. AB - The geochemical processes of mercury in Dongfeng (DF) and Wujiangdu (WJD) reservoirs, which were constructed in 1992 and 1979, respectively in Wujiang River, which is the upper branch of Yangtze River were investigated. One sampling site was chosen up river of 1 km from the dam for each reservoir. Three sampling campaigns were conducted at these sampling sites in December 2003, April 2004 and July 2004, respectively. The distributions of different mercury species in the water column, sediment, and sediment pore water were studied. We found that the sediment is the net source of both inorganic and MeHg to the water column for both reservoirs. The MeHg diffusion fluxes in WJD reservoir at all sampling campaigns were significantly higher than those in DF reservoir. Our study demonstrated that the high primary productivity in the reservoir produced elevated organic matter content that would favor the methylmercury production in sediment. PMID- 19564065 TI - Ageing processes and soil microbial community effects on the biodegradation of soil (13)C-2,4-D nonextractable residues. AB - The biodegradation of nonextractable residues (NER) of pesticides in soil is still poorly understood. The aim of this study was to evaluate the influence of NER ageing and fresh soil addition on the microbial communities responsible for their mineralisation. Soil containing either 15 or 90-day-old NER of (13)C-2,4-D (NER15 and NER90, respectively) was incubated for 90 days with or without fresh soil. The addition of fresh soil had no effect on the mineralisation of NER90 or of SOM, but increased the extent and rate of NER15 mineralisation. The analyses of (13)C-enriched FAME (fatty acids methyl esters) profiles showed that the fresh soil amendment only influenced the amount and structure of microbial populations responsible for the biodegradation of NER15. By coupling biological and chemical analyses, we gained some insight into the nature and the biodegradability of pesticide NER. PMID- 19564066 TI - Aniline and 2,4,6-trinitrotoluene associate preferentially to low molecular weight fractions of dissolved soil organic matter. AB - Aniline and 2,4,6-trinitrotoluene (TNT) were equilibrated with particulate (POM) and dissolved organic matter (DOM) from an organic soil at different compositions of adsorbed major cations (Na, Al) and pH (aniline: 3.7-5.1, TNT: 4.8-5.0). After separation of POM, concentrations of (14)C-labelled aniline and TNT* (including TNT degradation products) were determined in DOM size fractions using size exclusion chromatography (SEC) and UV-detection. Concentrations in the <3.5 kDa size fraction were 2.8-6.0 and 8.5-9.5 times higher for aniline and TNT*, respectively, as compared to the >40 kDa fraction. Thus, both aniline and TNT* were preferentially associated to the smallest DOM size fraction. The significant binding to DOM (similar extent as to POM) and the fact that the <3.5 kDa DOM fraction was less susceptible to flocculation by major metals suggests that the mobility of aniline and TNT is highly affected by the solubility of soil organic matter. PMID- 19564067 TI - Competitive biodegradation of dichlobenil and atrazine coexisting in soil amended with a char and citrate. AB - The role of char nutrients in the biodegradation of coexisting dichlobenil and atrazine in a soil by their respective bacterial degraders, DDN and ADP, was evaluated. Under growing conditions, their degradation in soil extract was slow with <40% and <20% degraded within 64 h, respectively. The degradation in extracts and slurries of char-amended solids increased with increasing char content, due to nutritional stimulation on microbial activities. By supplementing soil extract with various major nutrients, the measured degradation demonstrated that P was the exclusive limiting nutrient. The reduction in the degradation of coexisting dichlobenil and atrazine resulted apparently from the competitive utilization of P by DDN and ADP. With a shorter lag phase, ADP commenced growing earlier than DDN with the advantage of utilizing P first in insufficient supply. This resulted in an inhibition on the growth of DDN and thus suppression on dichlobenil degradation. PMID- 19564068 TI - Sedimentary record of hydrophobic organic compounds in relation to regional economic development: a study of Taihu Lake, East China. AB - Sediment cores taken from Taihu Lake, East China were analyzed for polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs) and polybrominated biphenyl ethers (PBDEs). The results showed a general sharp increase of HCH, DDT, PAH and PBDE concentrations in the surface layers, corresponding to a sedimentation time of 1980 and 1990 onward in the Meiliang Bay and Xukou Bay, respectively. The source of PAHs has largely transferred from petrogenic to pyrogenic origin, and good relationships were observed between sediment PAH concentrations and the regional gross domestic product. The sharp increase of DDTs in recent years may be related to the mobilization and migration of these chemicals from surface soil to lake sediment, as a result of enhanced soil run off due to large scale land transform, as well as the contribution of current usage of dicofol and DDT-containing anti-fouling paints. PMID- 19564069 TI - Environmental-geochemical characteristics of Cu in the soil and water in copper rich deposit area of southeastern Hubei Province, along the middle Yangtze River, Central China. AB - In this study, the natural Cu background concentration and Cu natural and anthropogenic contamination in soil, water and crop were investigated systematically in Huangshi area. The results show that regional geology is the dominant factor controlling the natural Cu background concentration in soil and water, and that pH is important to control the vertical distribution of Cu in soil under the same geographical and climatic conditions. The mineralization of rock bodies causes the natural Cu increase in soil and water, whereas, a large number of mining-smelting plants and chemical works are the main sources of Cu anthropogenic contamination. Cu in naturally and anthropogenically polluted soil displays differences in total and available contents, vertical distribution patterns and physico-chemical properties, the same happens in water. PMID- 19564071 TI - Elevated circulating soluble thrombomodulin activity, tissue factor activity and circulating procoagulant phospholipids: new and useful markers for pre-eclampsia? AB - OBJECTIVE: One of the most frequently proposed mechanisms for pre-eclampsia refers to uteroplacental thrombosis. However, the contribution of classical thrombotic risk factors remains questionable. The aims of this study were to investigate the activities of thrombomodulin, tissue factor and procoagulant phospholipids to assess endothelial cell injury in pregnant women with pre eclampsia and to compare them with other classical markers of vascular injury and thrombotic risk. STUDY DESIGN: Using three new functional assays we studied the plasma levels of these new markers in 35 healthy women, 30 healthy pregnant women, and 35 women with pre-eclampsia. RESULTS: We found that plasma levels of thrombomodulin activity, tissue factor activity and procoagulant phospholipids were significantly elevated in women with pre-eclampsia versus normal pregnant and non-pregnant women. CONCLUSION: It is thus suggested that elevated levels of these parameters in pre-eclampsia may reflect vascular endothelium damage, and may be a more valuable biomarker than antigen for the assessment of endothelial damage in pre-eclampsia. The high increased levels of procoagulant phospholipids and tissue factor activities in pre-eclampsia could suggest that the procoagulant potential may be implicated in this complication and makes these markers very promising for the understanding, follow-up and therapeutic handling of complicated pregnancy. PMID- 19564070 TI - A 3-year retrospective and clinical follow-up study of zirconia single crowns performed in a private practice. AB - OBJECTIVES: All-ceramic crowns have become increasingly used also in the posterior regions, but there are only few studies documenting the clinical outcome of these restorations. The aim was therefore to present the 3-year outcome of zirconia (NobelProcera) single crowns. METHODS: All patients treated with porcelain-veneered zirconia single crowns in a private practice during the period October 2004 to November 2005 were included. The patient records were scrutinized for data on the restorations and the supporting teeth. Information was available for 161 patients and 204 crowns. At the 3-year recall appointments, a sample of 18 patients with 25 crowns was clinically examined and interviewed regarding patient satisfaction. RESULTS: Most crowns (78%) were placed on premolars and molars. The clinical outcome of the crowns was favourable. No zirconia core fractured and no caries was observed on the abutment teeth. Some types of complication were recorded for 32 (16%) crowns or abutment teeth. The most severe complications, in total 12 restorations (6%), were recorded as failures: abutment tooth was extracted (5), remake of crown due to lost retention (4), veneer fracture (2) and persistent pain (1). The CDA criteria for 25 crowns were rated favourably, and patient satisfaction with the zirconia crowns was in general high. CONCLUSIONS: The porcelain-veneered zirconia crowns (NobelProcera) showed good clinical results, were well accepted by the patients, and only few complications were reported over the 3-year follow-up period. PMID- 19564072 TI - One laptop per child, local refurbishment or overseas donations? Sustainability assessment of computer supply scenarios for schools in Colombia. AB - With the intention of bridging the 'digital divide' many programmes have been launched to provide computers for educational institutions, ranging from refurbishing second hand computers to delivering low cost new computers. The fast and economical provision of large quantities of equipment is one of the many challenges faced by such programmes. If an increase is to be achieved in the sustainability of computer supplies for schools, not only must equipment be provided, but also suitable training and maintenance delivered. Furthermore, appropriate recycling has to be ensured, so that end-of-life equipment can be dealt with properly. This study has evaluated the suitability of three computer supply scenarios to schools in Colombia: (i) 'Colombian refurbishment', refurbishment of computers donated in Colombia, (ii) 'Overseas refurbishment', import of computers which were donated and refurbished abroad, and (iii) 'XO Laptop', -purchase of low cost computers manufactured in Korea. The methods applied were: Material Flow Assessment, -to assess the quantities-, Life Cycle Assessment, -to assess the environmental impacts, and the application of the Multiple Attribute Utility Theory, -to analyse, evaluate and compare different scenarios. The most sustainable solution proved to be the local refurbishment of second hand computers of Colombian origin to an appropriate technical standard. The environmental impacts of such practices need to be evaluated carefully, as second hand appliances have to be maintained, require spare parts and sometimes use more energy than newer equipment. Providing schools with second hand computers from overseas and through programmes such as 'One Laptop Per Child' has the disadvantage that the potential for social improvements - such as creation of jobs and local industry involvement - is very low. PMID- 19564073 TI - Suppression of cancer growth in mice by adeno-associated virus vector-mediated IFN-beta expression driven by hTERT promoter. AB - Adeno-associated virus (AAV) has rapidly become a promising gene delivery vehicle for its excellent advantages of non-immunogenic, low pathogenicity and long-term gene expression in vivo. However, a major obstacle in development of effective AAV vector is the lack of tissue specificity, which caused low efficiency of AAV transfer to target cells. The application of human telomerase reverse transcriptase (hTERT) promoter is a prior targeting strategy for AAV in cancer gene therapy as hTERT activity is transcriptionally upregulated in most cancer cells. In the present work, we investigated whether AAV-mediated human interferon beta (IFN-beta) gene driven by hTERT promoter could specifically express in tumor cells and suppress tumor cell growth. Our data demonstrated that hTERT promoter driven IFN-beta expression was the tumor-specific, decreased the cell viability of tumor cells but not normal cells, and induced tumor cell apoptosis via activation of caspase pathway and release of cytochrome c. AAV-mediated IFN-beta expression driven by hTERT promoter significantly suppressed the growth of colorectal cancer and lung cancer xenograft in mice and resulted in tumor cells death in vivo. These data suggested that AAVs in combination with hTERT-mediated IFN-beta expression could exert potential antitumor activity and provide a novel targeting approach to clinical gene therapy of varieties of cancers. PMID- 19564074 TI - Equilibrium and kinetic studies for sequestration of Cr(VI) from simulated wastewater using sunflower waste biomass. AB - In the present work, potential of sunflower head (BSH) [an agricultural waste biomass] to remove Cr(VI) from simulated wastewater has been evaluated under different process conditions such as pH, metal concentration, adsorbent dose and contact time. A contact time of 120 min and pH 2.0 were found to be optimum. Chromium removal decreased from 90.0 to 45.2% as its concentration increased from 10 to 70 mg/L. The Cr(VI) removal increased from 31.4 to 52.4% as adsorbent dose increased from 4.0 to 20.0 g/L using a test solution having 100mg/L Cr(VI) concentration. Adsorption isotherms were employed to evaluate the maximum adsorption capacity. Adsorption data fitted well to Langmuir isotherm and pseudo second order model as indicated by high values of correlation coefficient (0.9882 and 0.999) respectively. The adsorption capacity calculated from Langmuir isotherm was 8.177 mg/g. PMID- 19564075 TI - In-situ stabilization of Pb, Zn, Cu, Cd and Ni in the multi-contaminated sediments with ferrihydrite and apatite composite additives. AB - Three additives were evaluated for their effectiveness in the attenuation of Pb2+, Zn2+, Cu2+, Cd2+, Ni2+ in contaminated sediments. Apatite, ferrihydrite and their composite were applied to the sediments. For the remediation, BCR, SEM/AVS and TCLP were adopted as the evaluating method and comparison of their results were used for the first time to test in-situ stabilization effect. The results showed that after 5 months composite treatment, more than 70% Pb2+, 40% Zn2+, 90% Cu2+, 50% Cd2+ and 80% Ni2+ was immobilized in oxidizable and residual phases, respectively. Compared to untreated sediment, Pb2+, Zn2+, Cu2+, Cd2+ in residual fraction increased 20%, 10%, 10%, 10% with composite treatment after 5 months, respectively. SigmaSEM/AVS ratio declined from 12.6 to 9.3, in addition, composite treatments reduced the leaching of Pb2+ and Zn2+ from 10.6 mg L(-1) and 42.5 mg L(-1) to 5.4 mg L(-1) and 24.1 mg L(-1) in the sediment by TCLP evaluation. Meanwhile, apatite and ferrihydrite composite additives lowered the bioavailability and toxicity of sediments as well. Ferrihydrite had a positive effect in controlling the bioavailability and toxicity of heavy metals because it effectively retarded the oxidation of AVS in sediment. PMID- 19564076 TI - ZnFe2O4 nanoparticles: microwave-hydrothermal ionic liquid synthesis and photocatalytic property over phenol. AB - We report the microwave-hydrothermal ionic liquid (MHIL) synthesis and photocatalytic property over phenol of ZnFe(2)O(4) nanoparticles. Zn(CH(3)COO)(2).2H(2)O and Fe(NO(3))(3).9H(2)O were used as the zinc and iron sources, respectively, in the presence of CO(NH(2))(2) and the ionic liquid 1-n butyl-3-methyl imidazolium tetrafluoroborate ([BMIM][BF(4)]). Deionized water was used as a solvent. The ionic liquid [BMIM][BF(4)] and microwave heating temperature have significant influences on the crystal phase of the product. Different dosages of [BMIM][BF(4)] or microwave heating temperature could lead to the formation of different products such as ZnFe(2)O(4) and beta-FeOOH. The MHIL method has the advantages such as simplicity, rapidness and energy saving. The ZnFe(2)O(4) nanoparticles prepared by the MHIL method exhibit high photocatalytic activity for the degradation of phenol, which was up to 73% within 360 min. The TOC measurement confirmed the good photocatalytic efficiency of ZnFe(2)O(4) nanoparticles. PMID- 19564077 TI - Palliative care: Where have we come from and where are we going? PMID- 19564078 TI - An Item Response Theory (IRT) analysis of the Short Inventory of Problems-Alcohol and Drugs (SIP-AD) among non-treatment seeking men-who-have-sex-with-men: evidence for a shortened 10-item SIP-AD. AB - The Short Inventory of Problems-Alcohol and Drugs (SIP-AD) is a 15-item measure that assesses concurrently negative consequences associated with alcohol and illicit drug use. Current psychometric evaluation has been limited to classical test theory (CTT) statistics, and it has not been validated among non-treatment seeking men-who-have-sex-with-men (MSM). Methods from Item Response Theory (IRT) can improve upon CTT by providing an in-depth analysis of how each item performs across the underlying latent trait that it is purported to measure. The present study examined the psychometric properties of the SIP-AD using methods from both IRT and CTT among a non-treatment seeking MSM sample (N=469). Participants were recruited from the New York City area and were asked to participate in a series of studies examining club drug use. Results indicated that five items on the SIP AD demonstrated poor item misfit or significant differential item functioning (DIF) across race/ethnicity and HIV status. These five items were dropped and two parameter IRT analyses were conducted on the remaining 10 items, which indicated a restricted range of item location parameters (-.15 to -.99) plotted at the lower end of the latent negative consequences severity continuum, and reasonably high discrimination parameters (1.30 to 2.22). Additional CTT statistics were compared between the original 15-item SIP-AD and the refined 10-item SIP-AD and suggest that the differences were negligible with the refined 10-item SIP-AD indicating a high degree of reliability and validity. Findings suggest the SIP-AD can be shortened to 10 items and appears to be a non-biased reliable and valid measure among non-treatment seeking MSM. PMID- 19564079 TI - Recurrence cancer stem cells--made by cell fusion? AB - Within the past 10-15 years our knowledge about cancer and how cancer cells might originate has changed dramatically. It is now generally believed that a tumor has its origin in cancer stem cells (CSCs), which originated either from transformed tissue stem cells or transformed progenitor cells that have regained self-renewal activity. CSCs share several characteristics of normal stem cells, such as self renewal capacity, slow cell cycle activity, differentiation capacity, possessing an enhanced resistance towards cytotoxic agents and radiation, as well as tissue restoration capacity. Due to the increased drug and radiation resistance and slow cell cycle activity concomitant with tumor initiation capacity it is generally assumed that recurrent cancers originate from first line therapy surviving CSCs. But how does the CSC hypothesis explain "oncogenic resistance", which describes the phenomenon that most recurrent cancers are characterized by both an increased malignancy as well as resistance towards first line cancer therapy. To us, "oncogenic resistance" can not be simply attributed to the resistance properties of normal CSCs. If so, the recurring tumor should be treatable by first line therapy, which is mostly not the case. Thus, we conclude that "oncogenic resistance" demands a new type of tumor initiating cells, the so-called recurrence CSCs (rCSCs). This type of tumor initiating cell originates during first line therapy and is characterized by giving rise to first line therapy resistant and highly malignant progenies. Because several characteristics of "oncogenic resistance", such as increased drug resistance, increased resistance to apoptosis and an enhanced malignancy have been linked to cell fusion we further conclude that rCSCs might originate from this cellular event. However, which cell types have to fuse with each other to ultimately give rise to rCSCs is not clear. In any case, tumor tissues, particularly those being destructed by first line therapy comprise of a variety of fusogenic cells including tumor cells and CSCs as well as recruited monocytes/macrophages and bone marrow-derived stem cells. The fusogenic properties of these cells concomitant with phenotypic heterogeneity, which is also a property of cell fusion, will then lead to the origin of rCSCs. In accordance with Darwinian evolution only those cells will survive that can resist best to the selection pressure first line therapy. PMID- 19564080 TI - Can immunologic intercourse afford protection against renal transplant rejection? PMID- 19564081 TI - Artistic talent in dyslexia--a hypothesis. AB - The present article hints at a curious neurocognitive phenomenon of development of artistic talents in some children with dyslexia. The article also takes note of the phenomenon of creating in the midst of language disability as observed in the lives of such creative people like Leonardo da Vinci and Albert Einstein who were most probably affected with developmental learning disorders. It has been hypothesised that a developmental delay in the dominant hemisphere most likely 'disinhibits' the non-dominant parietal lobe to unmask talents, artistic or otherwise, in some such individuals. The present hypothesis follows the phenomenon of paradoxical functional facilitation described earlier. It has been suggested that children with learning disorders be encouraged to develop such hidden talents to full capacity, rather than be subjected to overemphasising on the correction of the disturbed coded symbol operations, in remedial training. PMID- 19564082 TI - Potential therapeutic advantages of guanosine over inosine in multiple sclerosis. PMID- 19564083 TI - Intestinal trefoil factor maybe useful in prophylaxis of acute graft-versus-host disease. AB - Graft-versus-host disease (GVHD) is a major complication of hematopoietic cell transplantation, GVHD pathophysiology can be divided into three phases, damage to the gastrointestinal (GI) tract in phase 1, principally by inflammatory cytokines, amplifies LPS release and leads to the "cytokine storm" characteristic of severe acute GVHD. It has been proved that disruption of phase 1 of the GVHD cascade is effective in prophylaxis of acute GVHD. Intestinal trefoil factor (ITF), a member of trefoil factor family (TFF) domain peptides, was proved to be very effective in prevention and healing of acute dextran sodium sulfate-induced colitis, and was also involved in protection against and recovery from intestinal mucositis induced by radiation and chemotherapy. So we hypothesise that ITF protects the intestinal tract mucosa from lesions and that it maybe useful in prophylaxis of acute GVHD. ITF can block GI tract damage in phase 1, preventing the amplification of the cascade. ITF may represent a novel strategy for the separation of GVHD and graft-versus-leukemia (GVL), and may serve as an effective adjunct to clinical regimens of GVHD prophylaxis. PMID- 19564084 TI - Distribution of antimicrobial resistance among different serovars of Haemophilus parasuis isolates. AB - The aim of the present study was to determine the antimicrobial susceptibility of Haemophilus parasuis in China and to investigate possible correlations between antimicrobial resistance and serovars or virulence. The susceptibility of 110 clinical isolates to 22 antimicrobial agents was determined by an agar dilution method. Except 28 isolates (25.5%) were non-typable, the isolates comprising thirteen serovars (1-5, 7-10, and 12-15), with the most prevalent were serovars 4 (14.5%), 5 (26.4%), 12 (9.1%) and 13 (10.9%). All isolates were susceptible to cefotaxime, ceftiofur, azithromycin, chloramphenicol, florfenicol and tilmicosin, and most of them were susceptible to penicillin, ampicillin, cefaclor, levofloxacin, ciprofloxacin, erythromycin, tetracycline, gentamicin, spectinomycin and tiamulin; however, a high level of resistance was encountered for enrofloxacin (70.9%) and trimethoprim/sulphamethoxazole (44.5%). There were 32 antimicrobial antibiograms observed, with 23.6% of the isolates resistant to more than 3 antimicrobial agents. Antimicrobial resistance was observed in every serovar included in this study, but no statistically significant association was observed between the antimicrobial resistance and the serovars or virulence of H. parasuis. PMID- 19564085 TI - Isolation of new anaerobic bacteria from sheep hooves infected with footrot. PMID- 19564086 TI - Cell population indexes of spermatogenic yield and testicular sperm reserves in adult jaguars (Panthera onca). AB - The intrinsic yield of spermatogenesis and supporting capacity of Sertoli cells are the desirable indicators of sperm production in a species. The objective of the present study was to quantify intrinsic yield and the Sertoli cell index in the spermatogenic process and estimate testicular sperm reserves by histological assessment of fragments obtained by testicular biopsy of five adult jaguars in captivity. The testicular fragments were fixed in 4% glutaric aldehyde, dehydrated at increasing alcohol concentrations, included into hydroxyethyl methacrylate, and were cut into 4 microm thickness. In the seminiferous epithelium of the jaguar, 9.2 primary spermatocytes in pre-leptotene were produced by "A" spermatogonia. During the meiotic divisions only 3.2 spermatids were produced by a primary spermatocyte. The general spermatogenic yield of the jaguar was about 23.4 cells and each Sertoli cell was able to maintain about 19.2 germ cells, 11 of them were round spermatids. In each seminiferous epithelium cycle about 166 million spermatozoa were produced by each gram of testicular tissue. In adult jaguars, the general spermatogenic yield was similar to the yield observed in pumas, greater than that observed for the domestic cat, but less compared to most domestic animals. PMID- 19564088 TI - Paucity of medical information held by patients in their homes for consultation by medical emergency crews. PMID- 19564089 TI - Juvenile polyp. PMID- 19564090 TI - Progressive rash after recent antibiotic exposure. PMID- 19564091 TI - Coffee grounds emesis: not just an upper GI bleed. AB - BACKGROUND: Acute upper gastrointestinal (UGI) bleeding manifests as hematemesis, melena, or hematochezia. Initial management is identical, irrespective of nasogastric (NG) aspirate results. Current practice includes early upper endoscopy. Significantly fewer high-risk bleeding lesions are found on endoscopy in patients with coffee grounds vs. bloody NG aspirates. OBJECTIVE: We present a case series to illustrate that patients with coffee grounds emesis (CGE) often have other unsuspected illnesses that may be overlooked due to preoccupation with the GI bleed. CASE REPORTS: A retrospective chart review of a series of 6 patients presenting with CGE and admitted for upper GI bleeding was performed. All 6 patients were hemodynamically stable at admission. NG lavage showed coffee grounds that cleared easily. None of the patients required blood transfusions during their hospital stay. Endoscopy in 3 of 6 patients failed to find any significant UGI lesions or stigmata of recent bleed. CONCLUSION: Although patients were admitted for UGI bleeding, the more significant associated diagnoses included acute myocardial infarction, urosepsis, small bowel obstruction, bilateral pulmonary emboli, and acute renal failure. Hemodynamically stable patients presenting with coffee grounds NG aspirate and no fall in hemoglobin/hematocrit should be evaluated for other non-GI bleeding-related conditions even as the GI bleed is being managed. PMID- 19564092 TI - Penile incarceration secondary to an S-shaped lead pipe: removal with Dremel Moto Tool. AB - BACKGROUND: Penile incarceration or strangulation is a urologic emergency. OBJECTIVES: Several techniques to remove metallic objects strangulating the penis are described in the literature. The method utilized depends on the severity of the incarceration and the tools that are readily accessible. Prompt action and resourcefulness, with expeditious removal, prevents organ ischemia and vascular or mechanical sequelae. CASE REPORT: We describe a case in which a Dremel Moto Tool was used to remove a lead pipe strangulating a penile shaft, after failure of the string technique. CONCLUSION: A hospital-based Emergency Medical Services and Rescue program is a valuable resource to provide the tools needed for management of penile strangulation. Features of safe removal, including protecting the tissues from heat damage and mechanical injury from the cutting blade, are described. PMID- 19564093 TI - Gene expression of the invasive phenotype of TNF-alpha-treated MCF-7 cells. AB - BACKGROUND AND OBJECTIVES: TNF-alpha secreted by tumor cells and macrophages that have infiltrated into the micro-environment of a tumor may promote the metastasis of a variety of malignant cancers, including breast cancer. The present study was designed to detect gene expression changes in metastatic MCF-7 cells treated with a low dose of TNF-alpha (20ng/mL), and to further explore the mechanisms by which TNF-alpha can contribute to metastasis. METHODS: Transwell assays were performed to evaluate the invasive phenotype of MCF-7 cells. Samples for cDNA array analysis were collected 3h and 24h after pre-treatment with TNF-alpha and changes in gene expression were quantitated. RESULTS: The invasive phenotype of MCF-7 cells was enhanced by the exposure of MCF-7 cells to a low dose of TNF-alpha. Gene expression profiles of 39 genes significantly increased or decreased after treatment with TNF-alpha, 6 of which were genes not previously associated with regulation by TNF-alpha. Genes to promote metastasis, as well as to inhibit metastasis, were identified with some changes being time dependent. CONCLUSIONS: TNF-alpha can enhance the invasive capacity of MCF-7 cells by affecting the expression of a group of genes that have roles in various steps of metastasis. Mechanistic insights into the role of TNF-alpha in tumor cell metastasis are discussed. PMID- 19564094 TI - Quality of life and non-pain symptoms in patients with cancer. AB - To measure the prevalence of non-pain physical symptoms and psychological symptoms in patients with cancer, to investigate the impact of physical and psychological symptoms on their quality of life (QoL), and to inquire whether treatment had been received for the complaints/symptoms, a representative sample of 1,429 cancer patients were recruited and classified according to tumor type and treatment status [i.e., (1a) curative treatment >6 months ago, (1b) curative treatment or =2 co-morbidities. Control population was also evaluated for the presence of same co-morbidities and compared with cases. The various significant co-morbidities included: migraine (cases: 25.6% vs. controls: 15.2%; p=0.02), anxiety (cases: 2.4% vs. controls: 0%; p=0.04), depression (case: 5.2% vs. controls: 0.4%; p=0.0009), sleep disturbances (case: 6.8% vs. controls: 0.4%; p=0.0002), neurocysticercosis (cases: 15.6% vs. controls: 0%; p=<0.001), pulmonary tuberculosis (cases: 3.6% vs. controls: 0%; p=0.002) and extra-pulmonary tuberculosis (cases: 2.8% vs. controls: 0.4%; p=0.03). Less common co-morbidities were hypertension, diabetes, osteoarthritis, asthma, hypothyroidism, and acid-peptic disease. CONCLUSION: In people living with epilepsy attending neurology OPD services, co-morbid illnesses were commonly observed compared to healthy controls. PMID- 19564097 TI - Optimal thresholds of early response to atypical antipsychotics: application of signal detection methods. AB - OBJECTIVE: Identify the optimal magnitude of response to antipsychotic medication at various early time points that best predicts subsequent non-response at 8 weeks. METHODS: Data were pooled from 5 randomized, double-blind clinical trials of atypical antipsychotics in the treatment of schizophrenia and related disorders (n=1137 moderately-to-severely ill; n=300 less than moderately ill). Signal detection methods (receiver-operating characteristic curves) were used to identify the optimal response threshold based on percent change from baseline on the PANSS total score at different early time points (Weeks 1-4) to predict subsequent 'non-response' at 8 weeks (i.e., not 'minimally improved', 'much improved' or 'remitted') while holding the false positive rate to a level of 30% or less. Analyses were implemented separately for patients with schizophrenia who differed on baseline illness severity. RESULTS: Using Area Under the Curve (AUC) >or=0.8 to define optimal discriminative ability at the earliest time point, the early response threshold in moderately-to-severely ill patients for predicting not 'minimally improved' was <15% reduction in PANSS total at Week 2, not 'much improved' was <23% at Week 2, and not 'remitted' was <26% at Week 4. Similarly, in less than moderately ill patients, the optimal early response threshold for predicting not 'minimally improved' was <12% reduction in PANSS total at Week 2, and not 'much improved' was <14% at Week 1. CONCLUSION: Specific thresholds of response were identified at early time points for predicting subsequent non response. Not attaining these early response thresholds may serve as important clinical markers of subsequent non-response to antipsychotic therapy. PMID- 19564095 TI - Off-label prescriptions in Italian hospices: a national survey. AB - Off-label prescription is part of routine care in palliative medicine, but no information is available about the situation in Italy. A cross-sectional observational survey was undertaken on all 66 Italian palliative care freestanding inpatient units to describe off-label prescriptions. Data were collected on 507 patients. Each prescribed drug was matched with Italian Marketing Authorization indications: 159 drugs off-label (4.5% of all prescribed drugs) for the stated indication were given to 128 patients (25.2%), and drugs unlicensed for subcutaneous injection were given to 147 patients (85.4% of all subcutaneous prescriptions, excluding insulin and heparin). The off-label prescriptions were subsequently evaluated by referring to the Italian National Pharmaceutical Formulary (INPF) and the British Palliative Care Formulary (PCF2). Although drugs are frequently prescribed off-label in inpatient palliative care units in Italy, this strategy was not always backed by clinical evidence, and in some cases, official/authoritative sources, such as INPF and PCF2 did not support it. Clinical trials and/or agreed international guidelines are needed to support the off-label use of the most widely prescribed drugs in palliation. PMID- 19564098 TI - The dose effect of human bone marrow-derived mesenchymal stem cells on epidermal development in organotypic co-culture. AB - BACKGROUND: A wealth of evidences have shown the participation and benefits of bone marrow-derived mesenchymal stem cells (BM-MSCs) in wound healing and skin tissue repair in vivo. However, their role in epidermal development and reconstitution is not clearly investigated. OBJECTIVE: Here we examine the quantitative effect of human BM-MSCs on epidermal regeneration in vitro. METHOD: Human keratinocytes and BM-MSCs are cultured at ratios from 0% to 100% on top of a fibroblast-embedded collagen gel in a three-dimensional organotypic co-culture model at an air-liquid interface up to 20 days and analyzed by histochemical and immunochemical staining of filaggrin, involucrin and keratin 10 on days 14 and 20. Human BM-MSCs were tracked with quantum dots in organotypic co-cultures. RESULTS: It was found that epidermal development is strongly influenced by the percentage of co-cultured BM-MSCs. A strong chemotactic effect between keratinocytes and MSCs was seen in the group with 50% of BM-MSCs, which resulted in an impaired epidermal development, whereas at a low percentage of BM-MSCs (10%), a stratified epidermal structure resembling native skin was established on day 14 of culture. Moreover, the immunostaining studies revealed that BM-MSCs in the low percentage (10%) participated in the basal periphery of reconstructed epidermis and a similar pattern characteristic of native epidermis was demonstrated in this experimental group, which was superior to all other experimental groups in terms of the thickness of stratum corneum and the expression profile of epidermal differentiation markers. CONCLUSION: This study indicates the advantage of using a new skin equivalent model incorporating a small fraction of MSCs to develop biologically useful tissues for maintaining homeostasis during skin regeneration and wound healing process. PMID- 19564099 TI - Effect of surface charge and agglomerate degree of magnetic iron oxide nanoparticles on KB cellular uptake in vitro. AB - We synthesized three types of magnetic iron oxide nanoparticles (MNPs), which were meso-2,3-dimercaptosuccinic acid (DMSA) coated MNPs (DMSA@MNPs, 17.3+/-4.8 nm, negative charge), chitosan (CS) coated MNPs (CS@MNPs, 16.5+/-6.1 nm, positive charge) and magnetic nanoparticles agglomerates, formed by electronic aggregation between DMSA@MNPs and CS (CS-DMSA@MNPs, 85.7+/-72.9 nm, positive charge) respectively. The interactions of these MNPs with Oral Squamous Carcinoma Cell KB were investigated. The results showed that cellular uptakes of MNPs were on the dependence of incubation time, nanoparticles concentration and nanoparticles properties such as surface charge, size, etc. The cellular uptake was enhanced with the increase of incubation time and nanoparticles concentration. Although all MNPs could enter to cells, we observed apparent differences in the magnitude of nanoparticles uptaken. The cellular uptake of CS-DMSA@MNPs by KB cells was the highest and that of DMSA@MNPs was the lowest among the three types of MNPs. The same conclusions were drawn via the reduction of water proton relaxation times T(2)(*), resulting from the different iron load of labeled cells using a 1.5T clinical MR imager. The finding of this study will have implications in the chemical design of nanomaterials for biomedical applications. PMID- 19564100 TI - A study of adsorption behavior of human serum albumin and ovalbumin on hydroxyapatite/chitosan composite. AB - In situ adsorption of human serum albumin (HSA) and ovalbumin (OVA) was real-time monitored by piezoelectric quartz crystal impedance (PQCI) technique to fully understand the initial cellular response on hydroxyapatite/chitosan (HAP/CS) composite. The PQCI parameters, such as resonant frequency (f), static capacitance (C(s)), and motional resistance (R(m)) were measured for investigating the kinetic adsorption behaviors of both proteins. The change in frequency shifts (Deltaf) depends on the amount of the adsorbed protein, and the change in motional resistance (DeltaR(m)) results from the microporosity variation of HAP/CS coating. The results show that the amount of the absorbed HSA is much greater than that of OVA on HAP/CS coating because of the unique construction of HSA as well as a flexible protein. Furthermore, Deltaf and DeltaR(m) data were fitted according to the kinetic exponential decay equations. It can be seen that there is only one adsorption process for OVA, but the absorption process for HSA is followed by a rearrangement process, and the former process is faster than the rearrangement process. Subsequently, the composite binding with proteins were demonstrated by the Fourier transform infrared (FTIR), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). PMID- 19564101 TI - Radiochemotherapy for bladder cancer. AB - Standard treatment for muscle-invasive bladder cancer is cystectomy. Multimodality treatment, including transurethral resection of the bladder tumour, radiation therapy, chemotherapy and deep regional hyperthermia, has been shown to produce survival rates comparable with those of cystectomy. With these programmes, cystectomy has been reserved for patients with incomplete response or local relapse. During the past two decades, organ preservation by multimodality treatment has been investigated in prospective series from single centres and co operative groups, with more than 1000 patients included. Five-year overall survival rates in the range of 50-60% have been reported, and about three quarters of the surviving patients maintained their bladder. Clinical criteria helpful in determining patients for bladder preservation include such variables as small tumour size (<5 cm), early tumour stage, a visibly and microscopically complete transurethral resection, absence of ureteral obstruction, and no evidence of pelvic lymph node metastases. On multivariate analysis, the completeness of transurethral resection of a bladder tumour was found to be one of the strongest prognostic factors for overall survival. Patients at greater risk of new tumour development after initial complete response are those with multifocal disease and extensive associated carcinoma in situ at presentation. Close co-ordination among all disciplines is required to achieve optimal results. Future investigations will focus on optimising radiation techniques, including all possibilities of radiosensitisation (e.g. concurrent radiochemotherapy, deep regional hyperthermia), and incorporating more effective systemic chemotherapy, and the proper selection of patients based on predictive molecular makers. PMID- 19564102 TI - Dietary habits of firstly admitted Spanish COPD patients. AB - BACKGROUND & AIMS: We described dietary habits in a Spanish sample of COPD patients and assessed its adequacy according to dietary recommendations, which so far have never been published. METHODS: 275 patients hospitalized for the first time for a COPD exacerbation in Spain answered a 122-item food frequency questionnaire on their last 2 years dietary habits. A Spanish food composition table was used to estimate nutrient intake. Adequacy of dietary intakes was assessed using the Spanish Nutrition Society guidelines for the elderly. RESULTS: Mean(SD) age was 68(8)years, 93% males, 32% current smokers, and post bronchodilator FEV(1) 53(16)% predicted. Mean(SD) body mass index was 28(5)kg/m2, with only 2% of subjects under 20kg/m2. Mean intake of meat, poultry and eggs was slightly higher (mean 108g/d) than recommended. Fish and shellfish (86g/d) as well as fruit and vegetables (567g/d) were adequate. Mean energy intake was 2033kcal/d. The proportion of energy from carbohydrates was lower (39%) whereas that from proteins and lipids were higher than recommended (20% and 35%, respectively). Alcohol intake was <30g in 80% of patients. Fatty acids, vitamin and mineral intakes were adequate, except for a deficiency in vitamin D (4.5microg/d). Stratification by socio-demographic and clinical characteristics showed very similar results. CONCLUSIONS: Moderate-to-severe Spanish COPD patients report an adequate intake of the main food groups and macro- and micro nutrients according to local recommendations, excepting vitamin D. PMID- 19564103 TI - Measuring organic carbon, nutrients and heavy metals in rivers receiving leachate from controlled and uncontrolled municipal solid waste (MSW) landfills. AB - Since landfilling is the common method of waste disposal in Malaysia, river water is greatly exposed to the risk of contamination from leachate unless proper leachate management is carried out. In this study, leachates from three different types of landfills, namely active uncontrolled, active controlled and closed controlled, were characterized, and their relationships with river water chemistry were examined monthly for a year. The influence of leachate on river water chemistry from each type of landfill depended on many factors, including the presence of a leachate control mechanism, leachate characteristics, precipitation, surface runoff and the applied treatment. The impact of leachate from an active uncontrolled landfill was the highest, as the organic content, NH(4)(+)-N, Cd and Mn levels appeared high in the river. At the same time, influences of leachate were also observed from both types of controlled landfills in the form of inorganic nitrogen (NH(4)(+)-N, NO(3)(-)-N and NO(2)(-)-N) and heavy metals (Fe, Cr, Ni and Mn). Improper treatment practice led to high levels of some contaminants in the stream near the closed controlled landfill. Meanwhile, the active controlled landfill, which was located near the coastline, was exposed to the risk of contamination resulting from the pyrite oxidation of the surrounding area. PMID- 19564104 TI - Oxidation of anthracene by immobilized laccase from Trametes versicolor. AB - The laccase of Trametes versicolor was immobilized on the functionalized nanoparticles SBA-15 with the average diameter less than 10 nm. Laccase mediated oxidations of anthracene (ANT) were investigated in the presence of two mediators, 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) and 1-hydroxybenzotriazole (HBT). Oxidation of ANT was more efficiently enhanced by adding 1 mM of HBT than that by adding ABTS. After 48 h oxidation HBT group significantly oxidized ANT with residue 58% relative to 88% in the ABTS group. HPLC and GC/MS analyses indicated the main product of ANT oxidation was anthraquinone (ANQ). The fluorescein diacetate (FDA) uptake of two human cell lines was used to assess the cytotoxicity and genotoxicity of ANT and ANQ. Treatments with ANT and ANQ at 5 and 10 microM exhibited significant cytotoxicity to the HaCaT cells and the A3 lymphocytes and no significant genotoxicity was observed. The results illustrated that ANQ is less toxic than ANT as well. PMID- 19564105 TI - Induced production of chitinase to enhance entomotoxicity of Bacillus thuringiensis employing starch industry wastewater as a substrate. AB - Induced production of chitinase during bioconversion of starch industry wastewater (SIW) to Bacillus thuringiensis var. kurstaki HD-1 (Btk) based biopesticides was studied in shake flask as well as in computer-controlled fermentors. SIW was fortified with different concentrations (0%; 0.05%; 0.1%; 0.2%; 0.3% w/v) of colloidal chitin and its consequences were ascertained in terms of Btk growth (total cell count and viable spore count), chitinase, protease and amylase activities and entomotoxicity. At optimum concentration of 0.2% w/v colloidal chitin, the entomotoxicity of fermented broth and suspended pellet was enhanced from 12.4x10(9) (without chitin) to 14.4x10(9) SBU/L and from 18.2x10(9) (without chitin) to 25.1x10(9) SBU/L, respectively. Further, experiments were conducted for Btk growth in a computer-controlled 15 L bioreactor using SIW as a raw material with (0.2% w/v chitin, to induce chitinase) and without fortification of colloidal chitin. It was found that the total cell count, spore count, delta-endotoxin concentration (alkaline solubilised insecticidal crystal proteins), amylase and protease activities were reduced whereas the entomotoxicity and chitinase activity was increased with chitin fortification. The chitinase activity attained a maximum value at 24 h (15 mU/ml) and entomotoxicity of suspended pellet reached highest (26.7x10(9) SBU/L) at 36 h of fermentation with chitin supplementation of SIW. In control (without chitin), the highest value of entomotoxicity of suspended pellet (20.5x10(9) SBU/L) reached at 48 h of fermentation. A quantitative synergistic action of delta-endotoxin concentration, spore concentration and chitinase activity on the entomotoxicity against spruce budworm larvae was observed. PMID- 19564106 TI - Highly enantioselective reduction of 4-(trimethylsilyl)-3-butyn-2-one to enantiopure (R)-4-(trimethylsilyl)-3-butyn-2-ol using a novel strain Acetobacter sp. CCTCC M209061. AB - The biocatalytic reduction of 4-(trimethylsilyl)-3-butyn-2-one to enantiopure (R) 4-(trimethylsilyl)-3-butyn-2-ol was successfully conducted with high enantioselectivity using immobilized whole cells of a novel strain Acetobacter sp. CCTCC M209061, newly isolated from kefir. Compared with other microorganisms that were investigated, Acetobacter sp. CCTCC M209061 was shown to be more effective for the bioreduction reaction, and afforded much higher yield and product enantiomeric excess (e.e.). The optimal buffer pH, co-substrate concentration, reaction temperature, substrate concentration and shaking rate were 5.0, 130.6 mM, 30 degrees C, 6.0 mM and 180 r/min, respectively. Under the optimized conditions, the maximum yield and the product e.e. were 71% and >99%, respectively, which are much higher than those reported previously. Additionally, the established biocatalytic system proved to be efficient for the bioreduction of acetyltrimethylsilane to (R)-1-trimethylsilylethanol with excellent yield and product e.e. The immobilized cells manifested a good operational stability under the above reaction conditions since they retained 70% of their catalytic activity after ten cycles of use. PMID- 19564107 TI - Synthesis and evaluation of 5-benzylidene(thio)barbiturate-beta-D-glycosides as mushroom tyrosinase inhibitors. AB - A series of 5-benzylidene(thio)barbiturate-beta-d-glycosides were designed, synthesized and evaluated as a new class of mushroom tyrosinase inhibitors. The results demonstrated that most of compounds had more potent inhibitory activities than arbutin (IC(50) 8.4mmol/L). Compound 12b was found to be the most potent inhibitor with IC(50) value of 0.05mmol/L. SARs analysis suggested that (1) 5 benzylidenethiobarbiturate substructures were efficacious for the inhibitory activity; (2) the lipophilic property of acetylated sugar moiety facilitated the inhibitory potency; (3) the hydroxyl group of 3'-configuration contributed to the increase of inhibitory effects. In addition, the inhibition mechanism study revealed that 5-benzylidene(thio)barbiturate-beta-d-glycosides were irreversible inhibitors. PMID- 19564108 TI - 4-(Phenylsulfonamidomethyl)benzamides as potent and selective inhibitors of the 11beta-hydroxysteroid dehydrogenase type 1 with efficacy in diabetic ob/ob mice. AB - Selective inhibitors of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) have considerable potential as treatments for type 2 diabetes. Presented herein are the syntheses, structure-activity relationships, and efficacy evaluation of 4 (phenylsulfonamidomethyl)benzamides as 11beta-HSD1 inhibitors. Through modification of our initial lead 5, we have identified potent and selective 11beta-HSD1 inhibitors, such as 11n, which demonstrated improved glycemic control, decreased serum lipids, and enhanced insulin sensitivity when dosed ip in diabetic ob/ob mice. PMID- 19564109 TI - Efficient synthesis of 3,3-diheteroaromatic oxindole analogues and their in vitro evaluation for spermicidal potential. AB - Syntheses of 3,3-diheteroaromatic oxindole derivatives has been achieved by coupling indole-2,3-dione (isatin) with differently substituted indoles and pyrrole in presence of I(2) in i-PrOH. The in vitro spermicidal potentials and the mode of spermicidal action of the synthesized analogues were evaluated and the derivative, 3,3-bis (5-methoxy-1H-indol-3-yl) indolin-2-one (3d) exhibited most significant activity. PMID- 19564110 TI - Identification of MK-1925: a selective, orally active and brain-penetrable opioid receptor-like 1 (ORL1) antagonist. AB - Structure-activity relationship studies directed toward improving the metabolic stability of compound 1 resulted in the identification of 3-[5-(3,5 difluorophenyl)-3-({[(1S,3R)-3-fluorocyclopentyl]amino}methyl)-4-methyl-1H pyrazol-1-yl]propanenitrile 39 (MK-1925) as a selective, orally available and brain-penetrable opioid receptor-like 1 (ORL1) antagonist. The compound also showed in vivo efficacy after oral dosing. Therefore, compound 39 was selected to undergo further studies as a clinical candidate. PMID- 19564111 TI - Acute and critical care management of a pediatric patient with medullo-cerebellar impaling. AB - The authors present a child with an accidental cervical medullo-cerebellar impaling by an aluminum rod. Careful planning for safe removal of the rod as well as vigilant attention to early cardiac instability and flash neurogenic pulmonary edema were paramount to her successful recovery. This patient illustrates that it is possible to survive impaling of the brainstem but it requires both innovation and collaboration by multiple specialists across different departments. The value of well coordinated and collaborative neuro surgical intensive care is demonstrated in this young girl's nearly complete recovery from the accident. PMID- 19564112 TI - Surgery for thoracic myelopathy caused by ossification of the ligamentum flavum. AB - Between January 1996 and December 2003, our department treated 16 patients (10 men and 6 women; average age 57.5 years) by performing a laminectomy for thoracic myelopathy caused by ossification of the ligamentum flavum (OLF). We followed up all patients for 36 to 86 months (mean follow-up time, 57.3 months). The mean (+/ standard deviation) Japanese Orthopaedic Association score increased from 5.0+/ 1.4 points before the operation to 7.7+/-1.9 points at the last follow-up (p<0.01). The average values for pre-operative and post-operative kyphosis of the involved vertebrae were 5.8 degrees +/-4.1 degrees and 8.8 degrees +/-6.0 degrees , respectively; the mean increase in kyphosis was only 3.0 degrees +/-2.4 degrees . An intraoperative dural tear was the main complication and none of the patients developed severe neurological complications. We conclude that laminectomy was both effective and safe in the treatment of thoracic OLF, but it must be performed with great care because of frequent dural adhesions to the OLF. The increase in kyphosis after the laminectomy was minimal when most of the facet joints were left intact and when the patient followed a back extensor exercise program post-operatively. PMID- 19564113 TI - Olfactory hallucinations as a manifestation of hidden rhinosinusitis. AB - The exact anatomical origin of olfactory epileptic seizures is controversial. Partial seizures can be an uncommon manifestation of brain tumours involving the medial temporal lobe and the orbitofrontal structures or as a marker of mesial temporal sclerosis. We present a patient suffering from olfactory hallucinations associated with inflammatory tissue in the anterior ethmoidal cells, in the right fronto-ethmoidal recess and in the maxillary sinus. Our report supports the opinion that the involvement of the olfactory bulb is crucial for developing so called uncinate fits and is independent of other causes. PMID- 19564114 TI - Auditory P300 event-related potential in tobacco smokers. AB - The aim of this study was to elucidate the chronic effects of tobacco smoking on the P300, a neurophysiological index of cognitive function. Prospective study participants were recruited from a family medicine polyclinic. We selected 32 right-handed smokers who had smoked more than 15 cigarettes per day, by inhalation, for more than 2 years. The control population consisted of 32 right handed, age-matched healthy individuals who had never smoked. Event-related potentials (ERPs) were recorded with the auditory "oddball" two-tone discrimination task. The data from the central (Cz) and frontal (Fz) electrodes were analyzed. The P300 and N1 amplitudes at Fz were lower in the study population compared to the control group. The early component of ERP, the measure of mental speed (N1) latency at Fz was prolonged in the study group compared to the controls, possibly because early cognitive processes such as sensory input or initial encoding of sensory information were delayed in this group. For those who smoke, a decreased N1 amplitude might indicate delayed information processing and possibly short-term memory disturbance. Thus, chronic tobacco smoking may produce prefrontal cognitive dysfunction. PMID- 19564115 TI - Anticoagulants and antiplatelet agents and the risk of development and recurrence of chronic subdural haematomas. AB - Seventy-one patients from northern Sweden were diagnosed with chronic subdural haematomas (CSDH) and treated at the Department of Neurosurgery at Umea University Hospital over 12 months. Fifty-four patients with CSDH had a history of head trauma (trauma group), while 17 patients had no previous head trauma (non trauma group). In the non-trauma group 71% of patients were treated with anticoagulants or antiplatelet aggregation agents (AAA) compared to 18% in the trauma group. Considering only AAA, 59% of the non-trauma patients were treated with these drugs versus 17% of patients in the trauma group. The recurrence rate for all patients was 17%. These findings confirm that the use of anticoagulants and AAA is over-represented in patients with non-traumatic CSDH. In our study, recurrence was not associated with previous use of anticoagulants or AAA. PMID- 19564116 TI - Peripheral neuromodulation for pain. AB - The past decade (1999-2009) has witnessed a dramatic increase in the use of electrical stimulation to treat chronic, intractable pain. The implantation of electrodes in close proximity to peripheral nerves, known as peripheral nerve stimulation, has been enthusiastically adopted by neurosurgeons and interventional pain specialists. The most common conditions treated with this technique are headache and complex regional pain syndromes. The potential application of peripheral neuromodulation to relatively common and frequently disabling conditions such as migraine and lower back pain represents an exciting phase in the evolution of contemporary pain surgery. We review the available evidence relating to the use of peripheral nerve stimulation for the treatment of medically refractory, chronic non-cancer pain in a variety of clinical situations, highlight the absence of randomised controlled studies, and emphasise the need for scientifically sound research in this field. PMID- 19564117 TI - Episodic dystonic spasm of the bilateral edge of the tongue during a coma following anoxic encephalopathy. PMID- 19564118 TI - Simultaneous anterior cruciate ligament reconstruction and opening wedge high tibial osteotomy: Report of four cases. AB - Four patients, aged 37-50 years, with chronic anterior cruciate ligament (ACL) deficiency, medial compartment osteoarthritis and varus deformity, underwent simultaneous arthroscopic ACL reconstruction and opening wedge high tibial osteotomy using the TomoFix fixation device and hydroxyapatite wedges. The simultaneous procedure allowed our patients to perform a full weight-bearing exercise at 4 weeks after surgery. At device removal and concomitant second-look arthroscopy, all patients had either a cyclops-like lesion or partial tears at the point of contact between the reconstructed ACL and intercondylar notch. Therefore, subsequent notchplasty or re-notchplasty was required. Because of the small number of patients, the results should be considered preliminary. Given our findings of graft morbidity caused by the corrected postoperative alignment, adequate intercondylar notchplasty should be performed at the initial operation. PMID- 19564119 TI - Synthesis of amyloid beta peptide 1-42 (E22Delta) click peptide: pH-triggered in situ production of its native form. AB - Amyloid beta peptide (Abeta) 1-42 is known to be involved in the onset of Alzheimer's disease (AD). We developed a click peptide of Abeta1-42 as a useful tool for AD research on the basis of an O-acyl isopeptide method. The click peptide quickly produced intact Abeta1-42 via a pH-dependent O-to-N intramolecular acyl migration (pH-click). Herein, a click peptide (26-O-acyl isoAbeta1-42 (E22Delta)) of a new mutant Abeta1-42 (E22Delta) was synthesized. The mutant click peptide was more water-soluble than Abeta1-42 (E22Delta). Moreover it quantitatively converted to the native peptide under physiological conditions (pH 7.4, 37 degrees C). CD analyses showed a conformational change from a random-coil structure of the click peptide to a beta-sheet structure of the in situ produced Abeta1-42 (E22Delta). This click peptide is a useful precursor of a mutant Abeta1-42 to establish an experiment system for investigating the properties of the mutant. PMID- 19564120 TI - Structural characteristics of the tendinous cord-papillary muscle junction in healthy and hypertensive rats. AB - Although the myotendinous junction (MTJ) of skeletal striated muscle is well known, more detailed studies regarding the structure of the cardiac MTJ are scarce. The objective of the present study was to investigate the morphological characteristics of the MTJ in hearts of healthy and hypertensive (SHR) female rats using histological, ultrastructural and three-dimensional (SEM) methods, as well as to evaluate the expression of vinculin by immunofluorescence. In the two groups, light microscopy showed branching tendinous cords and collagen bundles penetrating the apex of the finger-like projections of the papillary muscle. SEM analysis revealed an enlarged apex of the papillary muscle in SHR which was not observed in healthy animals. The loss of force transmission appears to be compensated by the amplified connection between the papillary muscle and valvular collagen. A large number of intercalated disks close to the fiber apex, small amounts of an amorphous intercellular substance and numerous vesicles were observed in SHR. In these animals, the expression of vinculin was more marked showing a regular distribution and a pattern of transverse striations along the sarcolemma. The presence of this protein in transverse bands suggests that vinculin surrounds myofibrils in the region of the Z band. Vinculin staining was also more marked in the region of the tendinous cord-papillary muscle junctions of SHR compared to control animals. Vinculin was quantified by electrophoresis and higher amounts of this protein were observed in SHR compared to control animals. PMID- 19564121 TI - Methodology for measuring conformation of solvent-disrupted protein subunits using T-WAVE ion mobility MS: an investigation into eukaryotic initiation factors. AB - The methodology developed in the research presented herein makes use of chaotropic solvents to gently dissociate subunits from an intact macromolecular complex and subsequently allows for the measurement of collision cross section (CCS) for both the recombinant (R-eIF3k) and solvent dissociated form of the subunit (S-eIF3k). In this particular case, the k subunit from the eukaryotic initiation factor 3 (eIF3) was investigated in detail. Experimental and theoretical CCS values show both the recombinant and solvent disrupted forms of the protein to be essentially the same. The ultimate goal of the project is to structurally characterize all the binding partners of eIF3, determine which subunits interact directly, and investigate how subunits may change conformation when they form complexes with other proteins. Research presented herein is the first report showing retention of solution conformation of a protein as evidenced by CCS measurements of both recombinant and solvent disrupted versions of the same protein. PMID- 19564122 TI - Initial experience with a new method of external polyester scaffolding for infrainguinal vein grafts. AB - OBJECTIVES: This study aims to evaluate the feasibility of external polyester scaffolding in infrainguinal bypass grafting when available vein material is suboptimal due to varicosity or dilatation. Primary objectives were short-term primary patency, assisted primary patency and secondary patency. Secondary objectives were to assess the rate of graft stenoses, infections and other adverse effects related to the use of external scaffolding. MATERIALS AND METHODS: A total of 50 consecutive patients were included in this prospective, multicentre, feasibility study from six centres. The indication for infrainguinal bypass was critical limb ischaemia (64%), severe claudication (34%) or popliteal aneurysm (2%). Indications for the use of the external scaffolding were varicosity of the vein graft, ectatic vein graft or the use of spliced vein grafts with segments of widely differing diameters. Duplex scanning of the graft was done perioperatively and at follow-up visits at 1, 3, 6 and 12 months after operation. RESULTS: Primary patency, assisted primary patency and secondary patency at 6 months were 82.3% (+/-SE 6.2%), 88.6% (+/-SE 4.8%) and 92.1% (+/-SE 4.4%), respectively. Six graft stenoses were detected in duplex surveillance. There were no infections related to polyester mesh. CONCLUSIONS: External scaffolding of infrainguinal vein grafts may be a promising innovation. Early results from this multicentre study show that polyester mesh is safe and feasible adjunct to peripheral revascularization enabling the use of otherwise non-optimal vein grafts with acceptable short-term patency. PMID- 19564123 TI - Morphometric data on the accessory ligament of the deep digital flexor tendon in the equine hindlimb. AB - Desmopathy of the accessory ligament of the deep digital flexor tendon (ALDDFT) in the hindlimb has recently been described as a cause of lameness in horses. However, there is limited morphometric data on this ligament. In the present study, the ALDDFT was carefully dissected in 165 hindlimbs that were collected from an abattoir. Length, width (lateral-to-medial) and thickness (plantar-to dorsal) of the ligament were measured. It was found that the ALDDFT may be absent in a minority of horses, but when it was present the general morphology was variable. The ALDDFT can be a single rectilinear structure ranging from slender or poorly developed to quite large and can be divided, in part or in its entirety, into two or three bundles. PMID- 19564124 TI - Cerebral palsy in eastern Denmark: declining birth prevalence but increasing numbers of unilateral cerebral palsy in birth year period 1986-1998. AB - The Cerebral Palsy Registry in eastern Denmark has been collecting cases using a uniform data sampling procedure since birth year 1979. Children are included by two child neurologists and an obstetrician. Information on pregnancy, birth, neonatal period, impairments and demographic data are registered. The total cerebral palsy birth prevalence has been significantly decreasing since the birth period 1983-1986 with 3.0 per 1000 live births until the period 1995-1998 with 2.1 per 1000 live births. The overall decrease was seen in preterm infants (<31 weeks) as well as in term infants and despite a simultaneous fall in perinatal and early neonatal mortality in the preterm group. Analysing the subtypes of CP we found a significant increase in the numbers as well as the rate of unilateral CP with a simultaneous fall in the numbers as well as the rate of bilateral CP. The explanation of this rise is not obvious. A change from bilateral periventricular lesions to unilateral is a possibility, but no major change in the neonatal handling could be documented. Regarding associated impairments, developmental delay/learning disabilities as well as motor function assessed by ability to walk (unassisted/assisted), both have changed toward higher percentage of children with unassisted walking and in need of special education. PMID- 19564126 TI - Energy and protein needs of cats for maintenance, gestation and lactation. AB - In the present investigation, data on the energy intakes and energy needs, as well as protein and fat accretion, of queens during pregnancy, during lactation and after lactation are given. Eleven adult cats were used as experimental animals. Data were collected during the fourth and seventh week of pregnancy, the second and sixth week of lactation and the second and sixth week after lactation. The cats were fed dry kitten food. During gestation and after lactation, all measurements were performed with respiration chambers. During lactation, balance trials without respiration chambers were performed. Body weight was measured and nitrogen, carbon and energy balances were calculated. From these, protein and fat accretion, as well as the metabolisable energy intake, was calculated. The weight gain during gestation was linearly independent of the number of kittens. During lactation, all cats lost weight; nevertheless, all cats except one were heavier 2 weeks after lactation than at mating. The energy intake of the cats during gestation was 1.8 times the maintenance requirement in the fourth week and two times maintenance requirement in the seventh week, and these energy intakes differed greatly among individuals. The energy intake of the cats during lactation was clearly higher than that recommended by National Research Council (NRC)(1), whereas the recommended protein intake in the second week of lactation was met. As the calculated protein balance was negative, the NRC recommendation for protein intake seems to be too low. In comparison to previous data, the cats showed a higher energy intake during lactation (median 502kJ/kgBW/d, second week lactation), and the weight loss was much lower. Further investigations on pregnant and lactating cats are necessary to complete the database. PMID- 19564125 TI - A vascular hamartoma arising from the cervical spine of a cat. AB - A 15-month-old cat presented for evaluation of worsening generalized proprioceptive ataxia. Computed tomography of the cervical spine revealed the presence of a compressive extradural bony mass involving the dorsal aspect of C1. Surgical exploration and debulking of the mass was performed. Histological evaluation of the mass revealed fibrovascular tissue consistent with a vascular hamartoma. This mass was deemed to be originating from the soft tissue associated with the C1 vertebra with subsequent bony proliferation. Surgical debulking of the mass resulted in complete resolution of clinical signs with no evidence of recurrence 2 years after surgery. PMID- 19564128 TI - Clinical expertise: Learning together through observed practice. PMID- 19564127 TI - The application of subspace preconditioned LSQR algorithm for solving the electrocardiography inverse problem. AB - Regularization is an effective method for the solution of ill-posed ECG inverse problems, such as computing epicardial potentials from body surface potentials. The aim of this work was to explore more robust regularization-based solutions through the application of subspace preconditioned LSQR (SP-LSQR) to the study of model-based ECG inverse problems. Here, we presented three different subspace splitting methods, i.e., SVD, wavelet transform and cosine transform schemes, to the design of the preconditioners for ill-posed problems, and to evaluate the performance of algorithms using a realistic heart-torso model simulation protocol. The results demonstrated that when compared with the LSQR, LSQR-Tik and Tik-LSQR method, the SP-LSQR produced higher efficiency and reconstructed more accurate epcicardial potential distributions. Amongst the three applied subspace splitting schemes, the SVD-based preconditioner yielded the best convergence rate and outperformed the other two in seeking the inverse solutions. Moreover, when optimized by the genetic algorithms (GA), the performances of SP-LSQR method were enhanced. The results from this investigation suggested that the SP-LSQR was a useful regularization technique for cardiac inverse problems. PMID- 19564129 TI - Changes in muscle fiber density following a stroke. AB - OBJECTIVES: Previous studies have revealed a selective functional loss of the large, high-threshold motor units in the paretic muscles after lesion of the upper motor neuron. We set out to study the degree and the time course of the reorganization of the motor units following a stroke. METHODS: Examinations were performed on 59 patients with a unilateral ischemic stroke in the territory of the middle cerebral artery, and on 42 healthy controls. The duration of hemiparesis ranged from 2 weeks to 48 months. The fiber density (FD) in the abductor digiti minimi muscle was determined by means of single-fiber electromyography on both the hemiparetic and the unaffected side in the patients, and unilaterally in the control subjects. RESULTS: The FD was increased on the hemiparetic side relative to the unaffected side and the control group. This change correlated with the severity of the clinical signs. The FD increased during the first 10 months following the stroke and subsequently remained stable. CONCLUSIONS: The process of reinnervation in the muscles takes place in the acute phase after stroke. These changes are related to the severity of the symptoms. SIGNIFICANCE: Our findings suggest that trans-synaptic degeneration of the spinal motor neurons occurs shortly after the lesion of the upper motor neurons. PMID- 19564130 TI - Seizure state detection of temporal lobe seizures by autoregressive spectral analysis of scalp EEG. AB - OBJECTIVE: To investigate a novel application of autoregression (AR) spectral techniques for seizure detection from scalp EEG. METHODS: EEGs were recorded from twelve patients with left temporal lobe epilepsy. The Burg maximum entropy AR method was applied to the signals from four electrodes near the epileptic focus for each patient, and the AR spectra were parameterized based on scalp EEG features described by a neurologist, thus mimicking clinical seizure identification. The parameters measured spectral peak power, sharpness, and location in a delta/low theta frequency range. An optimized nonlinear seizure detection index, which accounted for spatial and temporal persistence of behavior, was then calculated. RESULTS: Performance was optimized using recordings from two patients (315h, 18 seizures). For the remaining 10 patients (1624h, 83 seizures) results are presented as a Receiver Operating Characteristic graph, yielding an overall event-based true positive rate of 91.57% and epoch based false positive rate of 3.97%. CONCLUSIONS: Performance of the AR seizure identification method is comparable to other approaches. Techniques such as artifact removal are expected to improve performance. SIGNIFICANCE: There is a real potential for this seizure detection method to be of practical clinical use in long-term monitoring. PMID- 19564131 TI - Differential distribution of coherence between beta-band subthalamic oscillations and forearm muscles in Parkinson's disease during isometric contraction. AB - OBJECTIVE: Under rest condition, beta-band (13-30Hz) activity in patients with Parkinson's disease (PD) is prominent in the subthalamic nucleus (STN). However, the beta-band coupling between STN and muscle activity, its distribution and relation to motor symptoms remains unclear. METHODS: Using up to five electrodes, we recorded local field potentials (LFPs) above (zona incerta, ZI) and within the STN at different recording heights in 20 PD patients during isometric contraction. Simultaneously, we registered activity of the contralateral flexor and extensor muscle. We analysed LFP-EMG coherence to estimate coupling in the frequency domain. RESULTS: Coherence analysis showed beta-associated coupling in the ZI and STN with more significant LFP-EMG coherences in the STN. Coherence varied depending on the localisation of the LFP and muscles. We found significant difference between coherence of the extensor and the flexor muscle to the same LFP (p=0.045). CONCLUSIONS: We demonstrated that coherence between beta-band oscillations and forearm muscles are differentially distributed in the subthalamic region and between the forearm muscles in Parkinson's disease during isometric contraction. However, the significant LFP-EMG coupling did not associate with motor deficits in PD patients. SIGNIFICANCE: The differential distribution of beta-band activity in the STN highlights the importance of a topographically distinct therapeutic modulation. PMID- 19564132 TI - Commentary from the Italian Association of Sleep Medicine on the AASM manual for the scoring of sleep and associated events: for debate and discussion. AB - In 2007, the American Academy of Sleep Medicine (AASM) completed a new manual for the scoring of sleep and associated events. The AASM manual is divided into separate sections relative to the parameters reported for polysomnography. The present commentary, accomplished by a Task Force of the Italian Association of Sleep Medicine, focuses on sleep scoring data, arousal rules, movement and respiratory events. Comparisons with the previous Rechtschaffen and Kales system are detailed and a number of methodological weaknesses are pointed out. Major comments address the 30-s scoring epochs, the restrictive approach to arousals and EEG activating patterns, the incomplete quantification of motor events and the thresholds for the definition of hypopnea. Since the new AASM manual is an iterative process, proposals for discussion and re-examination of the agreed criteria with other national and international organizations are encouraged. PMID- 19564133 TI - Cdx genes, inflammation and the pathogenesis of Barrett's metaplasia. AB - Metaplasia is the conversion of one cell or tissue type to another and can predispose patients to neoplasia. Perhaps one of the best-known examples of metaplasia is Barrett's metaplasia (BM), a pathological condition in which the distal oesophageal epithelium switches from stratified squamous to intestinal type columnar epithelium. BM predisposes to oesophageal adenocarcinoma and is the consequence of long-term acid bile reflux. The incidence of BM and oesophageal adenocarcinoma has risen dramatically in recent years. A key event in the pathogenesis of BM is the induction of oesophageal CDX2 expression. Importantly, recent data reveal the molecular mechanisms that link inflammation in the development of Barrett's metaplasia, CDX2 and the progression to cancer. This review highlights the relationship between inflammation, metaplasia and carcinogenesis. PMID- 19564134 TI - Immunological cross-reactivity of mycobacterial topoisomerase I and divergence from other bacteria. AB - Mycobacterium smegmatis topoisomerase I exhibits several distinctive characteristics among all topoisomerases. The enzyme is devoid of Zn2+ fingers found typically in other bacterial type I topoisomerases and binds DNA in a site specific manner. Using polyclonal antibodies, we demonstrate the high degree of relatedness of the enzyme across mycobacteria but not other bacteria. This absence of cross-reactivity from other bacteria indicates that mycobacterial topoisomerase I has diverged from Escherichia coli and other bacteria. We have investigated further the immunological properties of the enzyme by raising a panel of monoclonal antibodies that recognises different antigenically active regions of the enzyme and binds it with widely varied affinity. Inhibition of a C terminal domain-specific antibody binding by enzyme-specific and non-specific oligonucleotides suggests the possibility of using these monoclonal antibodies to probe the structure, function and in vivo role of the enzyme. PMID- 19564135 TI - Beta-galactosylceramide alters invariant natural killer T cell function and is effective treatment for lupus. AB - NZB/W female mice spontaneously develop systemic lupus, an autoantibody mediated disease associated with immune complex glomerulonephritis. Natural killer (NK) T cells augment anti-dsDNA antibody secretion by NZB/W B cells in vitro, and blocking NKT cell activation in vivo with anti-CD1 mAb ameliorates lupus disease activity. In the current study, we show that beta-galactosylceramide reduces the in vivo induction of serum IFN-gamma and/or IL-4 by the potent NKT cell agonist alpha-galactosylceramide and reduces NKT cell helper activity for IgG secretion. Treatment of NZB/W mice with the beta-galactosylceramide ameliorated lupus disease activity as judged by improvement in proteinuria, renal histopathology, IgG anti-dsDNA antibody formation, and survival. In conclusion, beta galactosylceramide, a glycolipid that reduces the cytokine secretion induced by a potent NKT cell agonist ameliorates lupus in NZB/W mice. PMID- 19564137 TI - Label free capacitive immunosensor for detecting calpastatin--a meat tenderness biomarker. AB - An immunological capacitive biosensor for calpastatin was developed, optimized and applied for the analysis of meat extract samples. Anti-calpastatin antibody was immobilized on a gold electrode modified with a self-assembled monolayer of mercaptoundecanoic acid and Protein A from Staphylococcus aureus, and the obtained immunosensor was inserted as the working electrode in an electrochemical cell of a flow injection system. The dynamic range of the sensor was 20 to 160 ng/mL calpastatin. The electrode could be regenerated and re-used for more than 7 days with minimal reduction in sensitivity. For the analysis of real samples, the target analyte was extracted from the Longissimus dorsi muscle from beef carcasses directly after slaughtering. The extract was analyzed both with the developed immunosensor and microtiter plate ELISA, and a good correlation was obtained. However the immunosensor offers advantages of speed, simplicity, sensitivity and possibility for miniaturization over conventional assays for calpastatin quantification. PMID- 19564136 TI - Paraventricular nucleus mediates pressor response to noradrenaline injection into the dorsal periaqueductal gray area. AB - The dorsal periaqueductal gray area (dPAG) is involved in cardiovascular modulation. In a previous study, we reported that noradrenaline (NA) microinjection into the dPAG of rats caused pressor response that was mediated by vasopressin release. Vasopressin is synthesized by magnocellular neurons in the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. In the present study, we verified which nuclei mediated the cardiovascular response to NA as well as the existence of direct neural projection from the dPAG to hypothalamic nuclei. Then, we studied the effect of treating either PVN or SON with the nonselective synaptic blocker cobalt chloride (1mM) on the cardiovascular response to NA (15 nmol) microinjection into dPAG. Attempting to identify neural projections from dPAG to hypothalamic nuclei, we microinjected the neuronal tracer biotinylated-dextran-amine (BDA) into the dPAG and searched varicosity containing nerve terminals in the PVN and SON. Unilateral cobalt-induced inhibition of synapses in the SON did not affect the cardiovascular response to NA. However, unilateral inhibition of PVN significantly reduced the pressor response to NA. Moreover, cobalt-induced inhibition of synapses in both PVN blocked the pressor response caused by NA microinjected into the dPAG. Microinjection of BDA into the dPAG evidenced presence of varicosity-containing neuronal fibers in PVN but not in SON. The results from cobalt treatment indicated that synapses in PVN mediate the vasopressin-induced pressor response caused by NA microinjection into the dPAG. In addition, the neuroanatomical results from BDA microinjection into the dPAG pointed out the existence of direct neural projections from the dPAG site to the PVN. PMID- 19564138 TI - Ternary complexation of carvedilol, beta-cyclodextrin and citric acid for mouth dissolving tablet formulation. AB - The purpose of this study was to improve the solubility and dissolution rate of carvedilol by forming a ternary complex with beta-cyclodextrin and citric acid and to formulate its mouth-dissolving tablets. The rationale for preparing mouth dissolving tablet of carvedilol was to make the drug available in a soluble form in the mouth, which would facilitate its absorption from the buccal cavity. This would help to overcome its first-pass metabolism and thereby improve bioavailability. Phase solubility studies revealed the ability of beta cyclodextrin and citric acid to complex with carvedilol and significantly increase its solubility. Ternary complexation of carvedilol was carried out with beta-cyclodextrin and citric acid by physical mixing, kneading and spray drying methods and the prepared complexes were characterized by Fourier transform infra red spectroscopy, differential scanning calorimetry, powder X-ray diffractometry, scanning electron microscopy and complexation efficiency. The complex obtained by the spray drying method resulted in highest complexation efficiency and a 110 fold increase in the solubility of carvedilol. The mouth-dissolving tablets formulated using the spray dried complex with suitable excipients showed 100 % dissolution within five minutes. Accelerated stability studies of mouth dissolving tablets carried out as per ICH guidelines revealed that the tablets were stable. PMID- 19564139 TI - Preparation of pegylated nano-liposomal formulation containing SN-38: In vitro characterization and in vivo biodistribution in mice. AB - 7-Ethyl-10-hydroxy-camptothecin (SN-38), a metabolite of irinotecan x HCl, is poorly soluble in aqueous solutions and practically insoluble in most physiologically compatible and pharmaceutically acceptable solvents. Formulation of SN-38 in concentrated pharmaceutical delivery systems for parenteral administration is thus very difficult. Due to their biocompatibility and low toxicity, liposomes were considered for the delivery of SN-38. In this study, pegylated liposomes with distearoylphosphatidylcholine, distearoylphosphatidylethanolamine containing SN-38 were prepared and their characteristics, such as particle size, encapsulation efficiency, in vitro drug release and biodistribution, were investigated. The particle size of liposomes was in the range of 150--200 nm. The encapsulation efficiency and in vitro release rate of pegylated liposomes was higher than those of non-pegylated liposomes. As expected, the distribution of pegylated liposomes in body organs such as liver, kidney, spleen and lung was considerably lower than that of non pegylated liposomes. Also, their blood concentration was at least 50 % higher than that of non-pegylated liposomes. PMID- 19564140 TI - Synthesis of certain pyrrole derivatives as antimicrobial agents. AB - In an effort to establish new pyrroles and pyrrolo[2,3-d] pyrimidines with improved antimicrobial activity we report here the synthesis and in vitro microbiological evaluation of a series of pyrrole derivatives. A series of new 2 aminopyrrole-3-carbonitriles (1a-d) were synthesized from the reaction of benzoin, primary aromatic amines and malononitrile, from which a number of pyrrole derivatives (2a-d to 5a-d) and pyrrolo[2,3-d]pyrimidines (6a-d to 10a, d) were synthesized. The in vitro antimicrobial testing of the synthesized compounds was carried out against Gram-positive, Gram-negative bacteria and fungi. Some of the prepared compounds, [2-amino-1-(2-methylphenyl)-4,5-diphenyl-1H-pyrrole-3 carbonitriles (1b), 2-amino-3-carbamoyl-1-(3-methylphenyl)-4,5-diphenyl-1H pyrroles (2b), N-(3-cyano-1-(2-methylphenyl)-4,5-diphenyl-1H-pyrrol-2-yl) acetamides (3b), N-(3-cyano-1-(3-methylphenyl)-4,5-diphenyl-1H-pyrrol-2-yl) acetamides (3c), 2-amino-1-(4-methoxyphenyl)-4,5-diphenyl-3-tetrazolo-1H-pyrroles (5d), 7-(4-methoxyphenyl)-5,6-diphenyl-7H-pyrrolo [2,3-d]pyrimidin-4(3H)-ones (7d), 7-(3-methylphenyl)-5,6-diphenyl-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-thione (9b) and N-(7-(2-methylphenyl)-5,6-diphenyl-7H-pyrrolo[2,3-d] pyrimidine)-N-aryl amines (10a)] showed potent antimicrobial activity. PMID- 19564141 TI - Synthesis of coumarin heterocyclic derivatives with antioxidant activity and in vitro cytotoxic activity against tumour cells. AB - The aim of the present work was to synthesise coumarinyl heterocycles and to elucidate the potential role of these compounds as antioxidants and cytotoxic agents against Dalton's lymphoma ascites tumour cells (DLA) and Ehrlich ascites carcinoma cells (EAC). The synthesis of coumarin derivatives containing pyrazole, pyrazolone, thiazolidin-4-one, 5-carboxymethyl-4-thiazolidinone and 3-acetyl 1,3,4-oxadiazole ring is reported. 4-Methylcoumarinyl-7-oxyacetic acid hydrazide (1) reacted with arylazopropanes or hydrazono-3-oxobutyrate derivatives to form pyrazole (3a-c) and pyrazolone derivatives (5a-c). Heterocyclisation of Schiff's bases of 1 with thioglycolic acid, thiomalic acid or acetic anhydride afforded novel heterocyclic derivatives 4-thiazolidinones (7a-c), 5-carboxymethyl-4 thiazolidinones (8a-c) and oxadiazoles (9a-c), respectively. Some of the compounds showed promising antioxidant activity in vitro and cytotoxic activity against DLA cells and EAC cells. PMID- 19564142 TI - Development of matrix controlled transdermal delivery systems of pentazocine: In vitro/in vivo performance. AB - The present study aimed to develop hydroxypropyl methylcellulose based transdermal delivery of pentazocine. In formulations containing lower proportions of polymer, the drug released followed the Higuchi kinetics while, with an increase in polymer content, it followed the zero-order release kinetics. Release exponent (n) values imply that the release of pentazocine from matrices was non Fickian. FT-IR, DSC and XRD studies indicated no interaction between drug and polymer.The in vitro dissolution rate constant, dissolution half-life and pharmacokinetic parameters (C(max), t(max), AUC(s), t(1/2), Kel, and MRT) were evaluated statistically by two-way ANOVA. A significant difference was observed between but not within the tested products. Statistically, a good correlation was found between per cent of drug absorbed from patches vs. C(max) and AUC(s). A good correlation was also observed when per cent drug released was correlated with the blood drug concentration obtained at the same time point. The results of this study indicate that the polymeric matrix films of pentazocine hold potential for transdermal drug delivery. PMID- 19564143 TI - Improvement of physicomechanical properties of carbamazepine by recrystallization at different pH values. AB - The morphology of crystals has an appreciable impact role on the physicochemical properties of drugs. Drug properties such as flowability, dissolution, hardness and bioavailability may be affected by crystallinity behaviours of drugs. The objective of this study was to achieve an improved physicomechanical property of carbamazepine powder through recrystallization from aqueous solutions at different pH values. For this purpose, carbamazapine was recrystallized from aqueous solutions at different pH values (1, 7, 11). The morphology of crystals was investigated using scanning electron microscopy; X-ray powder diffraction (XRPD) was used to identify polymorphism; thermodynamic properties were analyzed using differential scanning calorimetery (DSC). Dissolution rate was determined using USP dissolution apparatus. Mechanical behavior of recrystallized carbamazepine powders was investigated by making tablets under different compaction pressure and measuring their hardness. SEM studies showed that the carbamazepine crystallization in different media affected the morphology and size of carbamazepine crystals. The shape of carbamazepine crystals changed from flaky or thin plate-like to needle shape. XRPD and DSC results ruled out any crystallinity changes occurring due to the temperature during recrystallization procedure or pH of crystallization media. The crushing strength of tablets indicated that all of the recrystallized carbamazepine samples had better compactiblity than the original carbamazepine powder. In vitro dissolution studies of carbamazepine samples showed a higher dissolution rate for carbamazepine crystals obtained from media with pH 11 and 1. Carbamazepine particles recrystallized from aqueous solutions of different pH values (all media) appeared to have superior mechanical properties to those of the original carbamazepine sample. PMID- 19564144 TI - Production variables affecting characteristics of pellets in melt pelletization with wax combination in a laboratory scale spheronizer. AB - The purpose of the study was to evaluate the suitability of laboratory scale spheronizer for the production of spherical pellets loaded with diltiazem hydrochloride by wax combination. The 1:1 combination of cetyl alcohol and hydrogenated castor oil, as low and high melting point waxes, were used. The various production variables affecting the different characteristics of pellets and the process efficiency were evaluated. Drug loaded pellets were evaluated for drug release in distilled water. Bowl temperature primarily affects the sphericity and adhesion of pellets to the bowl. Mass temperature has a pronounced effect on size, size distribution and sphericity of pellets. Wax concentration affects all characteristics of pellets but adhesion was least affected. The effect of these three variables can be compensated by optimizing the friction plate speed. It has been found that the highest yield of pellets (850--1400 microm) with maximum sphericity can be produced by using 45 degrees C bowl temperature, 52 degrees C mass temperature and 1400 rpm friction plate speed. PMID- 19564145 TI - Development and evaluation of gastroretentive norfloxacin floating tablets. AB - Floating matrix tablets of norfloxacin were developed to prolong gastric residence time, leading to an increase in drug bioavailability. Tablets were prepared by the wet granulation technique, using polymers such as hydroxypropyl methylcellulose (HPMC K4M, HPMC K100M) and xanthan gum. Tablets were evaluated for their physical characteristics, viz., hardness, thickness, friability, and mass variation, drug content and floating properties. Further, tablets were studied for in vitro drug release characteristics for 9 hours. The tablets exhibited controlled and prolonged drug release profiles while floating over the dissolution medium. Non-Fickian diffusion was confirmed as the drug release mechanism from these tablets, indicating that water diffusion and polymer rearrangement played an essential role in drug release. The best formulation (F4) was selected based on in vitro characteristics and was used in vivo radiographic studies by incorporating BaSO4. These studies revealed that the tablets remained in the stomach for 180 +/- 30 min in fasting human volunteers and indicated that gastric retention time was increased by the floating principle, which was considered desirable for the absorption window drugs. PMID- 19564146 TI - Synthesis of novel 1,3,4-oxadiazole derivatives and their biological properties. AB - A novel series of 2-[3-(4-bromophenyl)propan-3-one]-5-(substituted phenyl)-1,3,4 oxadiazoles (4a-n) have been synthesized from 3-(4-bromobenzoyl)propionic acid (3) with the aim to get better anti-inflammatory and analgesic agents with minimum or without side effects (ulcerogenicity). Compound 3 was reacted with several aryl acid hydrazides (2a-n) in phosphorous oxychloride to obtain the title compounds. Structures of the synthesized compounds were supported by means of IR, 1H NMR and mass spectroscopy. Title compounds were evaluated for their anti-inflammatory, analgesic, ulcerogenic and antibacterial activities. Antibacterial activity was expressed as the corresponding minimum inhibitory concentration (MIC). A fair number of compounds were found to have significant anti-inflammatory and analgesic activities, while a few compounds showed appreciable antibacterial activity. The newly synthesized compounds showed very low ulcerogenic action. The findings of the present study indicate that cyclization of the carboxylic group of 3 into novel 1,3,4-oxadiazole nucleus resulted in increased anti-inflammatory and analgesic activities with a significant decrease of ulcerogenic activity. PMID- 19564147 TI - Antioxidant activity of NSAID hydroxamic acids. AB - In the present study, seven hydroxamic acid derivatives of nonsteroidal anti inflammatory drugs (NSAIDs) (ibuprofen, fenoprofen, ketoprofen, indomethacin and diclofenac) were found to possess significant antioxidant, radical scavenging and metal chelating activities. The most active antioxidant and radical scavenger was N-methylhydroxamic acid of diclofenac (ANT = 88.0% and EC50 = 60.1 microg mL-1). The activity of the standard substance, butylated hydroxyanisole, in the two assays was ANT = 86.9% and EC50 = 18.8 microg mL-1, respectively. Ibuproxam was the strongest iron chelator among investigated hydroxamic acids (EC50 = 255.6 microg mL-1), yet significantly weaker than the standard substance, EDTA (EC50 = 29.1 microg mL-1). It seems that different mechanism is involved in metal chelating activity than in antioxidant and radical scavenging activity. Antioxidant and radical scavenging activities may be connected with conjugation of the nitrogen lone electron pair with the carbonyl group. On the other hand, more hydrophilic substances tend to be better iron chelators. PMID- 19564148 TI - Violations of Fitts' law in a ballistic task. AB - The authors explored changes in the postural preparation and movement times during jumps into targets of different sizes placed at different distances from the participant. Both movement and preparation times scaled with movement distance. Neither movement nor preparation time showed an effect of target size, although preparation time showed a tendency to increase for smaller targets. These observations show that the classical Fitts' law can be violated in tasks that involve a ballistic component. The data corroborate a hypothesis that Fitts' law originates at the level of movement planning. PMID- 19564149 TI - Perceiving affordances for aperture passage in an environment-person-person system. AB - The authors investigated the perception of affordances for aperture passage in an environment-person-person (E-P-P) system, which comprised an adult perceiver and a child as a companion. Perceivers were 8 large and 8 small female undergraduates and were companioned with 1 large and 1 small girl. The perceivers perceptually judged the minimum aperture width for the E-P-P system, and then the adult-child dyads (a pair of people) actually walked through to determine the system's actual minimum aperture width. Results demonstrated that perceivers precisely judged the action capabilities of an E-P-P system on the basis of the body-scaled information of each adult-child dyad. The findings extended the previous concept of affordances for an environment-person system to affordances for an E-P-P system. PMID- 19564150 TI - Identification and validation of urinary biomarkers for differential diagnosis and evaluation of therapeutic intervention in anti-neutrophil cytoplasmic antibody-associated vasculitis. AB - Renal activity and smoldering disease is difficult to assess in anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV) because of renal scarring. Even repeated biopsies suffer from sampling errors in this focal disease especially in patients with chronic renal insufficiency. We applied capillary electrophoresis coupled to mass spectrometry toward urine samples from patients with active renal AAV to identify and validate urinary biomarkers that enable differential diagnosis of disease and assessment of disease activity. The data were compared with healthy individuals, patients with other renal and non-renal diseases, and patients with AAV in remission. 113 potential biomarkers were identified that differed significantly between active renal AAV and healthy individuals and patients with other chronic renal diseases. Of these, 58 could be sequenced. Sensitivity and specificity of models based on 18 sequenced biomarkers were validated using blinded urine samples of 40 patients with different renal diseases. Discrimination of AAV from other renal diseases in blinded samples was possible with 90% sensitivity and 86.7-90% specificity depending on the model. 10 patients with active AAV were followed for 6 months after initiation of treatment. Immunosuppressive therapy led to a change of the proteome toward "remission." 47 biomarkers could be sequenced that underwent significant changes during therapy together with regression of clinical symptoms, normalization of C reactive protein, and improvement of renal function. Proteomics analysis with capillary electrophoresis-MS represents a promising tool for fast identification of patients with active AAV, indication of renal relapses, and monitoring for ongoing active renal disease and remission without renal biopsy. PMID- 19564151 TI - Forward programming of pluripotent stem cells towards distinct cardiovascular cell types. AB - AIMS: The proliferative potential of pluripotent stem cell-derived cardiomyocytes is limited, and reasonable yields for novel therapeutic options have yet to be achieved. In addition, various clinical applications will require the generation of specific cardiac cell types. Whereas early cardiovascular precursors appear to be important for novel approaches such as reseeding decellularized hearts, direct cell transplantation may require ventricular cells. Our recent work demonstrated that MesP1 represents a master regulator sufficient to induce cardiovasculogenesis in pluripotent cells. This led to our hypothesis that 'forward programming' towards specific subtypes may be feasible via overexpression of distinct early cardiovascular transcription factors. METHODS AND RESULTS: Here we demonstrate that forced expression of Nkx2.5 similar to MesP1 is sufficient to enhance cardiogenesis in murine embryonic stem cells (mES). In comparison to control transfected mES cells, a five-fold increased appearance of beating foci was observed as well as upregulated mRNA and protein expression levels. In contrast to MesP1, no increase of the endothelial lineage within the cardiovasculogenic mesoderm was observed. Likewise, Flk-1, the earliest known cardiovascular surface marker, was not induced via Nkx2.5 as opposed to MesP1. Detailed patch clamping analyses showed electrophysiological characteristics corresponding to all subtypes of cardiac ES cell differentiation in Nkx2.5 as well as MesP1 programmed embryoid bodies, but fractions of cardiomyocytes had distinct characteristics: MesP1 forced the appearance of early/intermediate type cardiomyocytes in comparison to control transfected ES cells whereas Nkx2.5 led to preferentially differentiated ventricular cells. CONCLUSION: Our findings show proof of principle for cardiovascular subtype specific programming of pluripotent stem cells and confirm the molecular hierarchy for cardiovascular specification initiated via MesP1 with differentiation factors such as Nkx2.5 further downstream. PMID- 19564152 TI - Molecular imaging of endothelial progenitor cell engraftment using contrast enhanced ultrasound and targeted microbubbles. AB - AIMS: Imaging methods to track the fate of progenitor cells after their delivery would be useful in assessing the efficacy of cell-based therapies. We hypothesized that contrast-enhanced ultrasound (CEU) using microbubbles targeted to a genetically engineered cell-surface marker on endothelial progenitor cells (EPCs) would allow the targeted imaging of vascular engraftment. METHODS AND RESULTS: Rodent bone marrow-derived EPCs were isolated, cultured, and transfected to express the marker protein, H-2Kk, on the cell surface. Non-transfected EPCs and EPCs transfected with either null plasmid or Firefly luciferase served as controls. Control microbubbles (MB(C)) and microbubbles targeted to H-2Kk expressed on EPCs (MB(H-2Kk)) were constructed. Binding of targeted microbubbles to EPCs was assessed in vitro using a parallel plate flow chamber system. CEU imaging of EPC-targeted microbubbles was assessed in vivo using subcutaneously implanted EPC-supplemented Matrigel plugs in rats. In flow chamber experiments, there was minimal attachment of microbubbles to plated control EPCs. Although numbers of adhered MB(C) were also low, there was greater and more diffuse attachment of MB(H-2Kk) to plated H-2Kk-transfected EPCs. Targeted CEU demonstrated marked contrast enhancement at the periphery of the H-2Kk transfected EPC-supplemented Matrigel plug for MB(H-2Kk,) whereas contrast enhancement was low for MB(C). Contrast enhancement was also low for both microbubbles within control mock-transfected EPC plugs. The signal intensity within the H-2Kk-transfected EPC plug was significantly greater for MB(H-2Kk) when compared with MB(C). CONCLUSION: Microbubbles targeted to a genetically engineered cell-surface marker on EPCs exhibit specific binding to EPCs in vitro. These targeted microbubbles bind to engrafted EPCs in vivo within Matrigel plugs and can be detected by their enhancement on CEU imaging. PMID- 19564153 TI - Multiple cardiac proteasome subtypes differ in their susceptibility to proteasome inhibitors. AB - AIMS: The proteasome is the proteolytically active core of the ubiquitin proteasome system, which regulates vital processes and which can cause various diseases when it malfunctions. Therefore, the proteasome has become an attractive target for pharmaceutical interventions. Inhibition of the cardiac proteasome by specific proteasome inhibitors has been shown to attenuate cardiac hypertrophy and ischaemia reperfusion injury of the heart. We have resolved the cardiac proteasome into its subtypes and have addressed the key question of how proteasome inhibitors affect single cardiac proteasomal subtypes. METHODS AND RESULTS: The 20S proteasome from rat heart was dissected into three different subpopulations (groups I-III), each comprising 4-7 different subtypes. The major group (group II) comprises standard proteasome subtypes; the two minor subpopulations (groups I and III) contain intermediate proteasome subtypes. All subtypes exhibit chymotrypsin-, trypsin-, and caspase-like activity but to different degrees. We have tested the effect of two common proteasome inhibitors on the chymotrypsin-like activity of all subtypes: 20-30 nmol/L MG132 caused 50% inhibition of all subtypes from groups I and II, whereas 100 nmol/L was necessary to affect group III subtypes to the same extent. However, another inhibitor, bortezomib (VELCADE), already used clinically, inhibited 50% of the activity of group III proteasome subtypes even below 20 nmol/L, a concentration showing almost no effect on group I and II proteasome subtypes. The caspase-like activity of group II proteasome subtypes was not affected by MG132 and was inhibited by bortezomib only at concentrations above 100 nmol/L. CONCLUSION: These data show that different inhibitors have differential inhibitory effects on the various cardiac proteasome subtypes. Different cardiac subtypes are inhibited by the same dose of proteasome inhibitor to a different extent. PMID- 19564154 TI - Polymyxin B identified as an inhibitor of alternative NADH dehydrogenase and malate: quinone oxidoreductase from the Gram-positive bacterium Mycobacterium smegmatis. AB - Tuberculosis is the leading cause of death due to a single infectious agent in the world and the emergence of multidrug-resistant strains prompted us to develop new drugs with novel targets and mechanism. Here, we screened a natural antibiotics library with Mycobacterium smegmatis membrane-bound dehydrogenases and identified polymyxin B (cationic decapeptide) and nanaomycin A (naphtoquinone derivative) as inhibitors of alternative NADH dehydrogenase [50% inhibitory concentration (IC(50)) values of 1.6 and 31 microg/ml, respectively] and malate: quinone oxidoreductase (IC(50) values of 4.2 and 49 microg/ml, respectively). Kinetic analysis on inhibition by polymyxin B showed that the primary site of action was the quinone-binding site. Because of the similarity in K(m) value for ubiquinone-1 and inhibitor sensitivity, we examined amino acid sequences of actinobacterial enzymes and found possible binding sites for L-malate and quinones. Proposed mechanisms of polymyxin B and nanaomycin A for the bacteriocidal activity were the destruction of bacterial membranes and production of reactive oxygen species, respectively, while this study revealed their inhibitory activity on bacterial membrane-bound dehydrogenases. Screening of the library with bacterial respiratory enzymes resulted in unprecedented findings, so we are hoping that continuing efforts could identify lead compounds for new drugs targeting to mycobacterial respiratory enzymes. PMID- 19564155 TI - Enzymatic characterization and comparison of various poaceae UDP-GlcA 4-epimerase isoforms. AB - UDP-alpha-D-galacturonic acid (UDP-GalA) is a key precursor for the synthesis of various bacterial and plant polysaccharides. UDP-glucuronic acid 4-epimerase (UGlcAE) catalyses the reversible conversion of UDP-alpha-D-glucuronic acid to UDP-GalA. UGlcAEs isolated from bacterial species have different biochemical properties when compared with the isoenzymes from the plant dicot species, Arabidopsis. However, little is known about the specificity of UGlcAE in Poaceae species. Therefore, we cloned and expressed in Escherichia coli several maize and rice UGlcAE genes, and compared their enzymatic properties with dicot homologs from Arabidopsis. Our data show that UGlcAE isoforms in different plant species have different enzymatic properties. For example, the Poaceae UGlcAE enzymes from rice and maize have significantly lower K(i) for UDP-xylose when compared with the Arabidopsis enzymes. The epimerases from different plant species are very specific and unlike their bacterial homolog in Klebsiella pneumoniae, can only use UDP-GlcA or UDP-GalA as their substrate. This study demonstrates that although members of the plant UGlcAE isoforms are highly conserved, the in vitro enzymatic activity of specific Poaceae isoform(s) may be regulated differently by specific nucleotide or nucleotide sugar. PMID- 19564156 TI - Crystal structure of hypothetical protein HP0062 (O24902_HELPY) from Helicobacter pylori at 1.65 A resolution. AB - The HP0062 gene encodes a small acidic protein of 86 amino acids with a theoretical pI of 4.6. The crystal structure of hypothetical protein HP0062 from Helicobacter pylori has been determined at 1.65 A by molecular-replacement method. The crystallographic asymmetric unit contains dimer, in which HP0062 monomer folds into a helix-hairpin-helix structure. The two protomers are primarily held together by extensive hydrophobic interactions in an antiparallel arrangement, forming a four helix bundle. Aromatic residues located at a or g position in the heptad leucine zipper are not major contributor required for HP0062 dimerization but important for the thermostability of this protein. PMID- 19564157 TI - Dynamic expression of peptidylarginine deiminase 2 in human monocytic leukaemia THP-1 cells during macrophage differentiation. AB - Peptidylarginine deiminases (PADs) consist of five enzymes which are widely distributed in human and rodent tissues. The two types of enzymes are found in human peripheral blood cells; PAD4 mainly in granulocytes and monocytes and PAD2 in lymphocytes and macrophages. Little is known about the regulation of PAD expression in macrophages. Here, we report that PAD2 is expressed in human monocytic leukaemia THP-1 cells during differentiation into macrophages by 12-O tetradecanoylphorbol-13-acetate. During this differentiation, the levels of PAD2 mRNA and protein increased concomitantly, indicating the transcriptional regulation of PAD2 gene expression in the cells. The treatment of THP-1-derived macrophages with calcium ionophore A23187 generated vimentin deimination and resulted in the disruption of vimentin filament organization. We discuss the possible role of vimentin deimination in cell physiology. PMID- 19564158 TI - Identification of radicals formed in the reaction mixture of bovine kidney microsomes with NADPH. AB - In order to explore the mechanism of myoglobinuric renal toxicity, detection and identification of free radicals was performed for the reaction mixtures of bovine kidney microsomes. EPR measurements showed prominent signals for the control reaction mixture containing 2.0 mg protein/ml bovine kidney microsomes, 5 mM NADPH, 0.1 M 4-POBN and 29 mM phosphate buffer (pH 7.4). Addition of myoglobin (Mb) to the control reaction mixture resulted in increase of EPR peak height. The result indicates that Mb enhances the radical formation. An HPLC-EPR measurement showed three peaks with retention times of 29.4 min (P(1)), 32.4 min (P(2)) and 46.6 min (P(3)). HPLC-EPR-MS analyses of P(1) and P(2) gave ions at m/z 282. The results show that 4-POBN/hydroxypentyl radical adducts form in the reaction mixture. An HPLC-EPR-MS analysis of P(3) gave ions at m/z 266, indicating that 4 POBN/pentyl radical adduct forms in the reaction mixture. PMID- 19564159 TI - Structural insights into the enzymatic mechanism of serine palmitoyltransferase from Sphingobacterium multivorum. AB - Serine palmitoyltransferase (SPT) is a key enzyme of sphingolipid biosynthesis and catalyses the pyridoxal 5'-phosphate (PLP)-dependent decarboxylative condensation reaction of l-serine with palmitoyl-CoA to generate 3 ketodihydrosphingosine. The crystal structure of SPT from Sphingobacterium multivorum GTC97 complexed with l-serine was determined at 2.3 A resolution. The electron density map showed the Schiff base formation between l-serine and PLP in the crystal. Because of the hydrogen bond formation with His138, the orientation of the Calpha-H bond of the PLP-l-serine aldimine was not perpendicular to the PLP-Schiff base plane. This conformation is unfavourable for the alpha-proton abstraction by Lys244 and the reaction is expected to stop at the PLP-l-serine aldimine. Structural modelling of the following intermediates indicated that His138 changes its hydrogen bond partner from the carboxyl group of l-serine to the carbonyl group of palmitoyl-CoA upon the binding of palmitoyl-CoA, making the l-serine Calpha-H bond perpendicular to the PLP-Schiff base plane. These crystal and model structures well explained the observations on bacterial SPTs that the alpha-deprotonation of l-serine occurs only in the presence of palmitoyl-CoA. This study provides the structural evidence that directly supports our proposed mechanism of the substrate synergism in the SPT reaction. PMID- 19564161 TI - Serum 99th centile values for two heart-type fatty acid binding protein assays. AB - BACKGROUND: We have previously demonstrated that heart-type fatty acid binding protein (H-FABP) is an independent prognostic marker for survival after acute coronary syndrome (ACS). This study aimed to define the 99th centile values for H FABP as determined with two different assays, and to study the relationship with age, gender and renal function. METHODS: H-FABP was measured on redundant routine outpatient samples using the MARKIT-M (Dainippon) and the Evidence Investigator (Randox) assays. RESULTS: Two hundred and forty-two subjects with Siemens Ultra TnI value <0.045 microg/L (99th centile) were studied. In all, 174 subjects had estimated glomerular filtration rate (eGFR) >60 mL/min. The 99th centile values for subjects with eGFR >60 mL/min for the Evidence Investigator H-FABP were 5.3 and 5.8 microg/L and for the MARKIT-M H-FABP were 8.3 and 9.1 microg/L in female and male subjects, respectively. There is an increase in H-FABP with age in subjects with normal renal function for both assays. Gender comparison showed no significant difference for either assay. Comparison of samples showed that subjects with eGFR <60 mL/min showed a median increase of 0.71 microg/L with Evidence Investigator assay and 1.09 microg/L with MARKIT-M assay compared with subjects with eGFR >60 mL/min. Calibration differences were confirmed by cross measurement of calibrators and recombinant H-FABP. CONCLUSIONS: We have defined the 99th centile values for H-FABP in a population of primary and secondary care outpatients that can be used to risk stratify patients with ACS. We have confirmed that H-FABP increases with renal dysfunction and age, but have not confirmed the gender difference previously reported. PMID- 19564160 TI - The beneficial endophyte Trichoderma hamatum isolate DIS 219b promotes growth and delays the onset of the drought response in Theobroma cacao. AB - Theobroma cacao (cacao) is cultivated in tropical climates and is exposed to drought stress. The impact of the endophytic fungus Trichoderma hamatum isolate DIS 219b on cacao's response to drought was studied. Colonization by DIS 219b delayed drought-induced changes in stomatal conductance, net photosynthesis, and green fluorescence emissions. The altered expression of 19 expressed sequence tags (ESTs) (seven in leaves and 17 in roots with some overlap) by drought was detected using quantitative real-time reverse transcription PCR. Roots tended to respond earlier to drought than leaves, with the drought-induced changes in expression of seven ESTs being observed after 7 d of withholding water. Changes in gene expression in leaves were not observed until after 10 d of withholding water. DIS 219b colonization delayed the drought-altered expression of all seven ESTs responsive to drought in leaves by > or = 3 d, but had less influence on the expression pattern of the drought-responsive ESTs in roots. DIS 219b colonization had minimal direct influence on the expression of drought-responsive ESTs in 32-d old seedlings. By contrast, DIS 219b colonization of 9-d-old seedlings altered expression of drought-responsive ESTs, sometimes in patterns opposite of that observed in response to drought. Drought induced an increase in the concentration of many amino acids in cacao leaves, while DIS 219b colonization caused a decrease in aspartic acid and glutamic acid concentrations and an increase in alanine and gamma-aminobutyric acid concentrations. With or without exposure to drought conditions, colonization by DIS 219b promoted seedling growth, the most consistent effects being an increase in root fresh weight, root dry weight, and root water content. Colonized seedlings were slower to wilt in response to drought as measured by a decrease in the leaf angle drop. The primary direct effect of DIS 219b colonization was promotion of root growth, regardless of water status, and an increase in water content which it is proposed caused a delay in many aspects of the drought response of cacao. PMID- 19564163 TI - Determination of fibroblast growth factor 23. AB - BACKGROUND: Fibroblast growth factor 23 (FGF-23) is a recently discovered hormone, which plays a key role in phosphate regulation. To investigate whether FGF-23 can be determined reliably, we validated the three available FGF-23 assays. METHODS: Currently, two intact FGF-23 assays (Kainos; Immutopics) and one C-terminal FGF-23 assay (Immutopics) are available. We determined intra- and inter-assay variation, linearity and matrix interference in these assays. Moreover, we compared assay results from healthy subjects with those of patient groups with expected high FGF-23 concentrations. RESULTS: Intra-assay variation was reasonably good in all three assays. Inter-assay variation and linearity were poor for the intact Immutopics assay but reasonable for both other assays. Immutopics assays gave best results in ethylenediaminetetraacetic acid (EDTA) plasma, while the Kainos assay showed comparable reproducibility in EDTA plasma and serum. Although the manual of the Kainos assay states that an automatic washing machine can be used, acceptable results were only found by manual washing. Patients with kidney disease and patients with hypophosphatemic osteomalacia had increased C-terminal FGF-23 concentrations compared with healthy controls. CONCLUSION: Two assays of reasonable quality are available for FGF-23, the intact FGF-23 assay (Kainos) provided proper attention is paid to the washing procedure, and the C-terminal assay (Immutopics). PMID- 19564164 TI - Remarks on the acute intermittent porphyria. PMID- 19564165 TI - Selective emotional processing deficits to social vignettes in schizophrenia: an ERP study. AB - Schizophrenia is associated with abnormalities in emotional processing and social cognition. However, it remains unclear whether patients show abnormal neurophysiological responses during fast, online appraisals of the emotional meaning of social information. To examine this question, event-related potentials (ERPs) were collected while 18 schizophrenia patients and 18 demographically matched controls evaluated 2-sentence vignettes describing negative, positive, or neutral social situations. ERPs were time locked to a critical word (CW) in the second sentence that conferred emotional valence. A late positivity effect to emotional (vs neutral) CWs was seen in both groups (in controls, to negative and positive CWs; in patients, to negative CWs only). However, the controls showed a greater late positivity effect to the negative and positive (vs neutral) CWs than the schizophrenia patients at mid-posterior (negative vs neutral) and at right posterior peripheral (positive vs neutral) sites. These between-group differences arose from reduced amplitudes of the late positivity to the negative and positive CWs in the patients relative to the controls; there was no difference between the 2 groups in the amplitude of the late positivity to the neutral CWs. These findings suggest that schizophrenia is associated with a specific neural deficit during the online evaluation of emotionally valent, socially relevant information. PMID- 19564169 TI - Are Americans feeling less healthy? The puzzle of trends in self-rated health. AB - Although self-rated health is proposed for use in public health monitoring, previous reports on US levels and trends in self-rated health have shown ambiguous results. This study presents a comprehensive comparative analysis of responses to a common self-rated health question in 4 national surveys from 1971 to 2007: the National Health and Nutrition Examination Survey, Behavioral Risk Factor Surveillance System, National Health Interview Survey, and Current Population Survey. In addition to variation in the levels of self-rated health across surveys, striking discrepancies in time trends were observed. Whereas data from the Behavioral Risk Factor Surveillance System demonstrate that Americans were increasingly likely to report "fair" or "poor" health over the last decade, those from the Current Population Survey indicate the opposite trend. Subgroup analyses revealed that the greatest inconsistencies were among young respondents, Hispanics, and those without a high school education. Trends in "fair" or "poor" ratings were more inconsistent than trends in "excellent" ratings. The observed discrepancies elude simple explanations but suggest that self-rated health may be unsuitable for monitoring changes in population health over time. Analyses of socioeconomic disparities that use self-rated health may be particularly vulnerable to comparability problems, as inconsistencies are most pronounced among the lowest education group. More work is urgently needed on robust and comparable approaches to tracking population health. PMID- 19564170 TI - The physical and mental health of Australian Vietnam veterans 3 decades after the war and its relation to military service, combat, and post-traumatic stress disorder. AB - The long-term health consequences of war service remain unclear, despite burgeoning scientific interest. A longitudinal cohort study of a random sample of Australian Vietnam veterans was designed to assess veterans' postwar physical and mental health 36 years after the war (2005-2006) and to examine its relation to Army service, combat, and post-traumatic stress disorder (PTSD) assessed 14 years previously (1990-1993). Prevalences in veterans (n = 450) were compared with those in the Australian general population. Veterans' Army service and data from the first assessments were evaluated using multivariate logistic regression prediction modeling. Veterans' general health and some health risk factors were poorer and medical consultation rates were higher than Australian population expectations. Of 67 long-term conditions, the prevalences of 47 were higher and the prevalences of 4 were lower when compared with population expectations. Half of all veterans took some form of medication for mental well-being. The prevalence of psychiatric diagnoses exceeded Australian population expectations. Military and war service characteristics and age were the most frequent predictors of physical health endpoints, while PTSD was most strongly associated with psychiatric diagnoses. Draftees had better physical health than regular enlistees but no better mental health. Army service and war-related PTSD are associated with risk of illness in later life among Australian Vietnam veterans. PMID- 19564172 TI - Nanotechnology in biomaterials. PMID- 19564171 TI - Acute leukemia and myelodysplasia after adjuvant chemotherapy for breast cancer: durable remissions after hematopoietic stem cell transplantation. AB - BACKGROUND: Although secondary acute leukemias and myelodysplasia are the known complications of adjuvant chemotherapy for breast cancer, the treatment outcome of these secondary malignancies is presently unclear. We examined the clinical and pathological features as well as the treatment results of a series of patients with acute leukemia/myelodysplasia arising after adjuvant chemotherapy for breast cancer. PATIENTS AND METHODS: Patients referred to our institution during a 5-year period for treatment of acute leukemia/myelodysplasia and who had received adjuvant chemotherapy for breast cancer are included. Leukemia-free survival for the whole group and for patients who underwent hematopoietic stem cell transplantation (HSCT) was estimated. RESULTS: Fifteen women (14 with acute leukemia and one with myelodysplasia) were identified. Seven of 15 patients had received an anthracycline, cyclophosphamide and a taxane. Ten patients developed acute leukemia/myelodysplasia with a latency period of 2 years or less from initiation of chemotherapy. Although mixed-lineage leukemia (MLL) rearrangement was the commonest chromosomal abnormality (8 of 15 patients), various other chromosomal abnormalities were also detected. Twelve of 15 patients underwent HSCT (11 allogeneic and one autologous). Eleven of these 12 patients who underwent HSCT were in remission at a median follow-up of 20.4 months (range 4.4 53.3 months). CONCLUSION: Durable remissions can be achieved in patients who develop acute leukemia/myelodysplasia secondary to adjuvant chemotherapy for breast cancer and are able to undergo allogeneic HSCT. Our results indicate that HSCT should be an early consideration in the management of such patients who are suitable candidates for the procedure. PMID- 19564173 TI - Cancer incidence and exposure to 4,4'-methylene-bis-ortho-chloroaniline (MbOCA). AB - AIMS: To monitor the occurrence of cancer in a recently defined cohort of UK workers engaged in the manufacture of polyurethane elastomers using 4,4' methylene-bis-ortho-chloroaniline. METHODS: A cohort of 308 male production workers from seven factories have been enumerated. All employees had a minimum of 12 months employment and were first employed at one of the participating factories in the period 1973-2000. Mortality and cancer incidence data for the period 1979-2007 were compared with expected values based on national rates. RESULTS: Mortality from all cancers combined was below the expected value [observed (Obs) 5, standardized mortality ratio (SMR) 68]. There was a single death from bladder cancer (SMR 560). The incidence of all cancers combined was also below expectation [Obs 9, standardized registration ratio (SRR) 77]. Site specific incidence was unexceptional except there was a non-significant excess of bladder cancer based on two cases (SRR 328). CONCLUSIONS: The findings for bladder cancer should be treated with caution as they relate to a relatively early period of follow-up and are based on very small numbers. PMID- 19564174 TI - Investigating the technical adequacy of curriculum-based measurement in written expression for students who are deaf or hard of hearing. AB - This study investigated the technical adequacy of curriculum-based measures of written expression (CBM-W) in terms of writing prompts and scoring methods for deaf and hard-of-hearing students. Twenty-two students at the secondary school level completed 3-min essays within two weeks, which were scored for nine existing and alternative curriculum-based measurement (CBM) scoring methods. The technical features of the nine scoring methods were examined for interrater reliability, alternate-form reliability, and criterion-related validity. The existing CBM scoring method--number of correct minus incorrect word sequences- yielded the highest reliability and validity coefficients. The findings from this study support the use of the CBM-W as a reliable and valid tool for assessing general writing proficiency with secondary students who are deaf or hard of hearing. The CBM alternative scoring methods that may serve as additional indicators of written expression include correct subject-verb agreements, correct clauses, and correct morphemes. PMID- 19564175 TI - Receptivity to Taboka and Camel Snus in a U.S. test market. AB - INTRODUCTION: The two largest U.S. cigarette manufacturers introduced Swedish style low-nitrosamine smokeless tobacco (snus) to several U.S. test markets in summer 2006. Since then, snus brands and test markets have proliferated. METHODS: This article assesses consumer response by analyzing data from the 2006 and 2007 Indiana Adult Tobacco Survey (IATS), a statewide telephone survey of 3,544 adults. During those years, the IATS included questions on awareness and trial of Camel Snus and Taboka. Analyses examined rates and predictors of awareness and trial statewide, and within the central Indiana test market. RESULTS: Nineteen percent of Indiana adults were aware of either Taboka or Camel Snus in 2006 and 2007. Estimates are larger (29%) for central Indiana and larger still (70%) for central Indiana smokers. Trial of snus, however, was very low (1.5% statewide), except among male smokers in central Indiana, 20% of whom are estimated to have tried it. Multivariate analyses showed that trial was more likely among men than women (odds ratio [OR] 13.85), residents of central Indiana than those farther from Indianapolis (OR 2.96), recipients than nonrecipients of tobacco promotions (OR 6.08), and those believing that smokeless tobacco is less harmful than cigarettes compared with those who believe it is equally or more harmful (OR 3.86). DISCUSSION: Results from this study suggest substantial initial interest in the new products among male smokers in this test market, especially those who receive promotional mailings from tobacco companies, which often include coupons for free or discounted products. PMID- 19564176 TI - Discrepancies in households and other stakeholders viewpoints on the food security experience: a gap to address. AB - This paper reports results from a case study on household food insecurity needs and the interventions that address them. It aimed at comparing households' perceptions on food insecurity experience and vulnerability to those of other stakeholders: community workers, programme managers and representatives from donor agencies. Semi-structured interviews with 55 households and 59 other stakeholders were conducted. Content analysis was performed, using a framework encompassing food sufficiency, characterization of household food insecurity and vulnerability of households to food insecurity. Overall, the results draw attention to a gap between households and the other stakeholders, where the later do not seem always able to assess the realities of food-insecure households. Other areas of divergences include: characteristics of food insecurity, relative importance of various risk factors related to food insecurity and the effectiveness of the community assistance to enhance the households' ability to face food insecurity. These divergent perceptions may jeopardize the implementation of sustainable solutions to food insecurity. Training of stakeholders for a better assessment of households' experience and needs, and systematic evaluation of interventions, appear urgent and highly relevant for an adequate response to households' needs. Collaboration between all stakeholders should lead to knowledge sharing and advocacy for policies dedicated to poverty reduction. PMID- 19564177 TI - Kidney function and the risk of cardiovascular disease. PMID- 19564178 TI - Kidney function and risk of cardiovascular disease and mortality in women: a prospective cohort study. AB - OBJECTIVE: To evaluate the association of kidney function with cardiovascular disease and mortality among apparently healthy women. DESIGN: Prospective cohort study. SETTING: Women's Health Study, United States. PARTICIPANTS: 27 939 female health professionals aged >or=45 who were free of cardiovascular disease and other major disease and who provided a blood sample at study entry. MAIN OUTCOME MEASURES: Time to cardiovascular disease (non-fatal stroke, non-fatal myocardial infarction, coronary revascularisation procedures, or death from cardiovascular cause), specific cardiovascular disease events, and all-cause mortality. End points were confirmed after review of medical records and death certificates. RESULTS: Glomerular filtration rate (GFR) was estimated with the abbreviated Modification of Diet in Renal Disease Study equation. At baseline, 1315 (4.7%) women had GFR <60 ml/min/1.73 m(2). During 12 years of follow-up, 1199 incident cardiovascular disease events and 856 deaths (179 from cardiovascular disease) occurred. Compared with women with GFR >or=90 ml/min/1.73 m(2), the multivariable adjusted hazard ratios for any first cardiovascular disease were 0.95 (95% CI 0.83 to 1.08), 0.84 (0.70 to 1.00), and 1.00 (0.79 to 1.27) among women with GFR of 75-89.9, 60-74.9, and <60 ml/min/1.73 m(2), respectively; the equivalent hazard ratios for all cause mortality were 0.93 (0.79 to 1.09), 1.03 (0.85 to 1.26), and 1.09 (0.83 to 1.45). Similar null findings were observed for myocardial infarction, stroke, coronary revascularisation, and non-cardiovascular death. However, an increased risk of death from cardiovascular disease was found among women with GFR <60 ml/min/1.73 m(2) (hazard ratio 1.68 (1.02 to 2.79)). CONCLUSIONS: In this large cohort of women, a glomerular filtration rate <60 ml/min/1.73 m(2) was associated with increased risk of cardiovascular disease death but not other cardiovascular disease events or non-cardiovascular disease mortality. We observed no increase in risk of any of the outcomes among women with less severe impairment of kidney function. PMID- 19564179 TI - Multiple imputation for missing data in epidemiological and clinical research: potential and pitfalls. PMID- 19564180 TI - Breaking the mould in patient safety. PMID- 19564181 TI - Drug prevention of hypertension. Law's flaw? PMID- 19564182 TI - Drug prevention of hypertension. All cause absolute benefit? PMID- 19564183 TI - Drug prevention of hypertension. Best use of resources? PMID- 19564184 TI - Drug prevention of hypertension. Assumptions and extrapolation. PMID- 19564185 TI - Drug prevention of hypertension. Up to a point. PMID- 19564186 TI - Blood pressure in primary care. Home monitoring, a way to go? PMID- 19564188 TI - Time to carotid endarterectomy. Don't forget amaurosis fugax. PMID- 19564189 TI - Future of female doctors. Time to adapt the workplace. PMID- 19564191 TI - Erythromycin in whooping cough. PMID- 19564192 TI - Priorities in stroke care. The London stroke strategy. PMID- 19564194 TI - Celibacy of Catholic priests. Points of information. PMID- 19564195 TI - Experts suggest ways to tackle shortage of sperm donors. PMID- 19564196 TI - Celibacy of Catholic priests. Time for root and branch reform. PMID- 19564197 TI - Urgent action is needed to prevent deaths among Himalayan pilgrims, says climbing federation. PMID- 19564198 TI - Don't use urine microscopy to confirm microscopic haematuria. PMID- 19564200 TI - Collaborations aim to develop new drugs for neglected tropical diseases. PMID- 19564201 TI - Stimulating blood supply before ovary transplantation improves success. PMID- 19564203 TI - Donor pharmacological hemodynamic support is associated with primary graft failure in human heart transplantation. AB - The aim of this study was to test the impact of donor and recipient characteristics on the development of primary graft failure (PGF) after heart transplantation (HT) by focusing on the donor's inotropic support. Heart donors and matched recipients data were prospectively collected. Univariate and multivariate analyses were used to determine independent predictors for PGF and peri-operative mortality. The donor's high inotrope requirement was defined as sustained need for dopamine exceeding 10 microg/kg/min and/or alpha agonists exceeding 0.06 microg/kg/min. PGF instead was defined as need for immediate post HT mechanical circulatory support. Since 2006, we have performed 37 HTs. PGF occurred in six patients (16.2%). Although four patients (66.6%) were weaned off circulatory support, two of them (33.3%) died on mechanical assistance. Total in hospital mortality was 10.8% (4/37). Upon multivariate analysis, pre-harvesting donor high inotrope dosage was the major determinant for PGF (P=0.03, OR=10.8). Given the organ shortage, many centers accepted marginal hearts assuming the donor's pre-harvest hemodynamic managing has a reduced impact on PGF development. As PGF remains the most lethal postoperative complication, the hazards should be carefully considered when using pre-harvesting high inotrope infusion rates. PMID- 19564204 TI - Experimental selective sympathicotomy (ramicotomy) and sympathetic regeneration. AB - Ramicotomy is a surgical procedure, with less adverse effects than conventional sympathectomy, however, it was abandoned due to the high recurrence rate. Twenty eight pigs underwent bilateral videothoracoscopic ramicotomy and were divided into five groups. The animals were sacrificed at 15th, 45th, 90th, 135th and 180th postoperative days (POD). The segments were removed and evaluated for macroscopic regeneration and histological analysis. The data were compared to the control group of 10 intact segments of the sympathetic. There was no macroscopic regeneration on the 15th POD, and present on 41.6% on the 180th POD (P<0.05). The Schwann cells presented a similar evolution in both rami beginning at the 45th POD, with a smaller count in the gray rami. The collagen and reticular fibers presented a negative correlation (r=-0.414; P<0.01). The deposition of the collagen fibers was greater in the gray rami with a peak deposition on the 135th POD and a diminishing rate in the 180th POD (P<0.05). Ramicotomy allows complete section of all rami communicantes of the sympathetic ganglia. The histological regeneration might be greater than the recurrence rates of clinical symptoms seen in a human being due to non-functional regenerations. PMID- 19564205 TI - Early and mid-term functional and survival benefits in ischaemic versus degenerative mitral valve repair using Duran flexible ring: a single surgeon series. AB - The late results of ischaemic mitral valve (MV) repair have been less than satisfying. We compared echocardiographically, the changes in LV function, mid term durability and survival between MV repair caused by ischaemic cardiomyopathy (n=60) with degenerative MV disease (n=73) over a period of 15 years. The duration of mean follow-up was 3.7+/-4.1 years in the ischaemic group and 3.9+/ 2.9 years in the degenerative group. Freedom from reoperation at seven years was 98.3%+/-1.5% and 98.9%+/-2.1%, respectively (P=0.889). At the last follow-up, NYHA functional class I or II was present in 78.4% of patients in the ischaemic group and 80.9% patients in the degenerative group (P=0.347). An improvement in LVEF was noted in both the groups: ischaemic--41.3+/-12.7 (pre-op LVEF: 38.8+/ 14.1) and degenerative--46.5+/-11.1 (pre-op LVEF: 45.7+/-11.7) (P=0.014). At seven years, freedom from a cardiac cause of death was statistically similar in the two groups: 93.3%+/-1.3% and 92.2%+/-0.6% (P=0.967). In conclusion, the mid term results of ischaemic MV repair are similar to those obtained for degenerative MV repair. Surgical correction of ischaemic MR results in long-term improved LVEF and comparable outcomes in terms of freedom from reoperation and survival. PMID- 19564206 TI - Circulating lipoprotein(a) concentrations and abdominal aortic aneurysm presence. AB - To summarize the present evidence for an association between circulating lipoprotein(a) [Lp(a)] and abdominal aortic aneurysm (AAA) presence. MEDLINE database was searched to identify all case-control studies that compared circulating Lp(a) concentrations between patients with AAA and subjects without AAA. For each study, data regarding Lp(a) concentrations in both the AAA and control groups were used to generate standardized mean differences (SMDs) and 95% confidence intervals (CIs). Study-specific estimates were combined using inverse variance-weighted average of logarithmic SMDs in both fixed- and random-effects models. Our search identified five case-control studies. In total, our meta analysis included data on 982 cases with AAA and 1296 controls without AAA. Pooled analysis of the five studies demonstrated significantly higher Lp(a) concentrations in the AAA group than those in the control group in random-effect models (SMD, 0.26; 95% CI, 0.08-0.44; P=0.005). There was significant study heterogeneity of results (P=0.03) but no evidence of significant publication bias (P=0.3272). We found that circulating Lp(a) concentrations are likely higher in cases with AAA than in controls without AAA. Higher circulating Lp(a) concentrations may be associated with AAA presence. PMID- 19564207 TI - Prevention of arterial graft spasm by botulinum toxin: an in-vitro experiment. AB - In coronary artery bypass surgery, arterial grafts result in improved patency rates. However, these grafts frequently fail due to spasm. Papaverine has been used to prevent graft spasm, but its effect is short-lived. Botulinum toxin inhibits muscle contraction for about three months. We investigated the usefulness of botulinum toxin in preventing arterial grafts spasm in vitro. Samples of abdominal aorta from male Wistar rats were cut into 2 mm rings and treated with various doses of botulinum toxin or papaverine for 30 min. All rings were stimulated with KCl and noradrenaline. Tension was recorded using myography. We compared constriction caused by noradrenaline or KCl in rings treated with botulinum toxin, or papaverine, or physiological salt solution (PSS) (control). In the presence of KCl and noradrenaline, almost all concentrations of botulinum toxin completely inhibited arterial contraction when compared with controls. Spasm prevention was lost after 60 min in rings with papaverine but persisted for 120 min in rings with botulinum toxin. In the histological examination, arterial wall structure was not destroyed by botulinum toxin. Botulinum toxin prevented arterial graft spasm in vitro and had a longer lasting effect than papaverine, with no toxic effect on the artery. PMID- 19564208 TI - Ultrasonic scalpel for sealing of the thoracic duct: evaluation of effectiveness in an animal model. AB - To verify the usefulness of an ultrasonic scalpel for the sealing of lymphatic ducts, a Harmonic Ace scalpel (Ethicon Endo-Surgery, Inc, Cincinnati, OH) was tested, using the thoracic ducts in pigs. Indocyanine green was injected into the abdominal lymphatic ducts in two pigs, and the stained thoracic ducts were identified. The thoracic ducts were then divided and sealed into seven sections with a Harmonic Ace scalpel, used at a power setting of level 3. The cut ends of the thoracic ducts were evaluated macroscopically, histologically, and by measuring bursting pressure. The whole length of thoracic duct of each pig was clearly imaged by pigment. Each stump divided by the ultrasonic scalpel was completely sealed, and there was no pigment leakage from the cut end macroscopically. Histologic examination revealed that the cut end was sealed by homogenous degenerative coagulation. Bursting pressures could be determined at three cut ends of thoracic ducts and were 195 mmHg, 188 mmHg, and 203 mmHg, respectively. The thoracic ducts were reliably divided and sealed by an ultrasonic scalpel in pigs. This device is expected to be a useful tool for the surgical treatment of chylothorax. PMID- 19564209 TI - Gender differences in trajectories of health limitations and subsequent mortality. A study based on the German Socioeconomic Panel 1995-2001 with a mortality follow-up 2002-2005. AB - OBJECTIVES: Although research on health limitations has investigated gender differences in health and mortality, gender differentials in individual-level trajectories have been studied less frequently. Moreover, there are no studies on the relationship between course types and subsequent mortality. We investigate course types, explore confounding by socioeconomic and demographic correlates, and pose the question of whether the gender gap in morbidity results from differences in the onset of, and/or survival with, health limitations. METHODS: Using the German Socioeconomic Panel, we identify individual trajectories of health limitations and use multinomial logistic regressions to explore confounding and the relationship with mortality. RESULTS: The frequency of stable trajectories without limitations is lower among women because they tend to experience courses that involve extended periods of limitations and deteriorating health. Women also experience more frequently improvement after deterioration. The female mortality advantage is particularly large after health deterioration. DISCUSSION: Health limitations do not make men and women more equal in the face of death. Our results are consistent with earlier studies showing that mortality selection and differences in chronic conditions may explain the gender gap in health and mortality. We extend previous research showing that the female health disadvantage is largely the result of their mortality advantage. PMID- 19564211 TI - Haplotype-sharing analysis showing Uyghurs are unlikely genetic donors. AB - The Uyghur (UIG) are a group of people primarily residing in Xinjiang of China, which is geographically located in Central Asia, from where modern humans were presumably spread in all directions reaching Europe, east, and northeast Asia about 40 kya. A recent study suggested that the UIG are ancestry donors of the East Asian (EAS) gene pool. However, an alternative hypothesis, that is, the UIG is an admixture population with both EAS and EUR ancestries is also supported by our previous studies. To test the two competing hypotheses, here we conducted a haplotype-sharing analysis (HSA) based on empirical and simulated data of high density single nucleotide polymorphisms. Our results showed that more than 95% of UIG haplotypes could be found in either EAS or EUR populations, which contradicts the expectation of the null models assuming that UIG are donors. Simulation studies further indicated that the proportion of UIG private haplotypes observed in empirical data is only expected in alternative models assuming that UIG is an admixture population. Interestingly, the estimated ancestry contribution of 44%:56% (EAS:EUR) based on HSA is consistent with our previous estimation with STRUCTURE analysis. Although the history of UIGs could be complex, our method is explicit and conservative in rejecting the null hypothesis. We concluded that the gene pool of modern UIGs is more likely a sole recipient with contribution from both EAS and EUR. PMID- 19564210 TI - Caregiver-recipient closeness and symptom progression in Alzheimer disease. The Cache County Dementia Progression Study. AB - Applying Rusbult's investment model of dyadic relationships, we examined the effect of caregiver-care recipient relationship closeness (RC) on cognitive and functional decline in Alzheimer's disease. After diagnosis, 167 participants completed up to six visits, observed over an average of 20 months. Participants were 64% women, had a mean age of 86 years, and mean dementia duration of 4 years. Caregiver-rated closeness was measured using a six-item scale. In mixed models adjusted for dementia severity, dyads with higher levels of closeness (p < .05) and with spouse caregivers (p = .01) had slower cognitive decline. Effect of higher RC on functional decline was greater with spouse caregivers (p = .007). These findings of attenuated Alzheimer's dementia (AD) decline with closer relationships, particularly with spouse caregivers, are consistent with investment theory. Future interventions designed to enhance the caregiving dyadic relationship may help slow decline in AD. PMID- 19564212 TI - Cumulative and antagonistic effects of a mixture of the antiandrogens vinclozolin and iprodione in the pubertal male rat. AB - Vinclozolin and iprodione are dicarboximide fungicides that display antiandrogenic effects in the male rat, which suggests that a mixture would lead to cumulative effects on androgen-sensitive end points. Iprodione is a steroid synthesis inhibitor, but androgen receptor antagonist activity, which is displayed by vinclozolin, has not been fully evaluated. Here, we demonstrate that iprodione binds to the human androgen receptor (IC(50) = 86.0 microM), reduces androgen-dependent gene expression, and reduces androgen-sensitive tissue weights in castrated male rats (Hershberger assay). Since vinclozolin and iprodione affect common targets in the pubertal male rat, we tested the hypothesis that a mixture would have cumulative antiandrogenic effects. An iprodione dose, that does not significantly affect androgen-dependent morphological end points, was combined with vinclozolin doses (2 x 5 factorial design). Sprague-Dawley rats were dosed by gavage with vinclozolin at 0, 10, 30, 60, and 100 mg/kg/day with and without 50 mg iprodione/kg/day from postnatal day (PND) 23 to 55-57 (n = 8 per group). The age at puberty (preputial separation [PPS]), organ weights, serum hormones, and ex vivo testis steroid hormone production were measured. Vinclozolin delayed PPS, reduced androgen-sensitive organ weights, and increased serum testosterone. The addition of iprodione enhanced the vinclozolin inhibition of PPS (PND 47.5 vs.49.1; two-way ANOVA: iprodione main effect p = 0.0002). The dose response for several reproductive and nonreproductive organ weights was affected in a cumulative manner. In contrast, iprodione antagonized the vinclozolin-induced increase in serum testosterone. These results demonstrate that these fungicides interact on common targets in a tissue-specific manner when coadministered to the pubertal male rat. PMID- 19564213 TI - Domoic acid induces a long-lasting enhancement of CA1 field responses and impairs tetanus-induced long-term potentiation in rat hippocampal slices. AB - Domoic acid (DOM) is known to cause hippocampal neuronal damage and produces amnesic effects. We examined synaptic plasticity changes induced by DOM exposure in rat hippocampal CA1 region. Brief bath application of DOM to hippocampal slices produces a chemical form of long-term potentiation (LTP) of CA1 field synaptic potentials. The potentiation cannot be blocked by NMDA receptor antagonist MK-801 but can be blocked by the calcium-calmodulin-dependent protein kinase II (CaMKII) inhibitor KN-62 or cAMP-dependent protein kinase (PKA) inhibitor H-89. DOM-potentiated slices show decreased autophosphorylated CaMKII (p-Thr286), an effect that is also dependent on the activity of CaMKII and PKA. Increased phosphorylation of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor subunit GluR1 (p-Ser831) was seen in DOM-potentiated slices. Therefore, aberrant regulation of CaMKII and GluR1 phosphorylation occurs after DOM application. In addition, tetanus-induced LTP as well as the increase of phosphorylation of CaMKII (p-Thr286) were reduced in DOM-potentiated slices. Compared with brief exposure, slices recovering from prolonged exposure did not show potentiation or altered levels of CaMKII (p-Thr286) or GluR (p-Ser831). However, decreased phosphorylation of GluR1 at Ser845 was seen. These results describe a new chemical form of LTP and uncover novel molecular changes induced by DOM. The observed impairment of tetanus LTP and misregulation of CaMKII and GluR1 phosphorylation may partially account for DOM neurotoxicity and underlie the molecular basis for DOM-induced memory deficit. PMID- 19564214 TI - Contented but not better: problems with satisfaction. PMID- 19564215 TI - Congress restores the Americans With Disabilities Act to its original intent. AB - The Americans With Disabilities Act (ADA) was created to prohibit discrimination based on disability. Although many individuals filed claims alleging discrimination in the workplace based on disability, the federal courts, led by the U.S. Supreme Court, adopted an increasingly constricted interpretation of key provisions of the ADA. As a result, many individuals with serious impairments were found by the courts to not be disabled. In 2008 Congress overturned these court decisions by enacting the ADA Amendments Act. These amendments overrule several decisions by the United States Supreme Court that narrowed the applicability of the ADA, and they restore the ADA to its original intent. PMID- 19564216 TI - What is "women-focused" treatment for substance use disorders? AB - Over the past three decades research has highlighted gender differences in substance use disorders and substance abuse treatment participation. Programs devoted to addressing women's treatment needs, broadly encompassed in the term "women-focused treatment," have multiplied. This column examines the rationale for women-focused treatment and describes some of its components. The authors cite the need to evaluate women-focused treatment by developing validated measures of the processes embodied in such treatment and by conducting empirically sound research on clinical outcomes, treatment effectiveness, cost effectiveness, and the optimal means of providing services to women with substance use disorders. PMID- 19564217 TI - A national action plan for workforce development in behavioral health. AB - Across all sectors of the behavioral health field there has been growing concern about a workforce crisis. Difficulties encompass the recruitment and retention of staff and the delivery of accessible and effective training in both initial, preservice training and continuing education settings. Concern about the crisis led to a multiphased, cross-sector collaboration known as the Annapolis Coalition on the Behavioral Health Workforce. With support from the Substance Abuse and Mental Health Services Administration, this public-private partnership crafted An Action Plan for Behavioral Health Workforce Development. Created with input from a dozen expert panels, the action plan outlines seven core strategic goals that are relevant to all sectors of the behavioral health field: expand the role of consumers and their families in the workforce, expand the role of communities in promoting behavioral health and wellness, use systematic recruitment and retention strategies, improve training and education, foster leadership development, enhance infrastructure to support workforce development, and implement a national research and evaluation agenda. Detailed implementation tables identify the action steps for diverse groups and organizations to take in order to achieve these goals. The action plan serves as a call to action and is being used to guide workforce initiatives across the nation. PMID- 19564218 TI - Bipolar disorder center for Pennsylvanians: implementing an effectiveness trial to improve treatment for at-risk patients. AB - OBJECTIVE: Adolescents, elderly persons, African Americans, and rural residents with bipolar disorder are less likely than their middle-aged, white, urban counterparts to be diagnosed, receive adequate treatment, remain in treatment once identified, and have positive outcomes. The Bipolar Disorder Center for Pennsylvanians (BDCP) study was designed to address these disparities. This report highlights the methods used to recruit, screen, and enroll a cohort of difficult-to-recruit individuals with bipolar disorder. METHODS: Study sites included three specialty clinics for bipolar disorder in a university setting and a rural behavioral health clinic. Study operations were standardized, and all study personnel were trained in study procedures. Several strategies were used for recruitment. RESULTS: It was possible to introduce the identical assessment and screening protocol in settings regardless of whether they had a history of implementing research protocols. This protocol was also able to be used across the age spectrum, in urban and rural areas, and in a racially diverse cohort of participants. Across the four sites 515 individuals with bipolar disorder were enrolled as a result of these methods (69 African Americans and 446 non-African Americans). Although clinical characteristics at study entry did not differ appreciably between African Americans and non-African Americans, the pathways into treatment differed significantly. CONCLUSIONS: Rigorous recruitment and assessment procedures can be successfully introduced in different settings and with different patient cohorts, thus facilitating access to high-quality treatment for individuals who frequently do not receive appropriate care for bipolar disorder. PMID- 19564220 TI - The REACT study: cost-effectiveness analysis of assertive community treatment in north London. AB - OBJECTIVE: Assertive community treatment (ACT) is a key component of mental health care, but recent information on its cost-effectiveness is limited. This article provides a cost-effectiveness analysis of assertive community treatment and usual care from community mental health teams (CMHTs) in the United Kingdom. METHODS: Participants who had difficulties engaging with community services were randomly assigned to ACT (N=127) or continued usual care from CMHTs (N= 124). Costs were measured at baseline and 18 months later and compared between the two groups. In the analysis, cost data were linked to information on satisfaction, which had been shown to be significantly higher with ACT. RESULTS: Total follow up costs over 18 months were higher for the ACT group by pounds sterling 4,031 ($6,369), but this was not statistically significant (95% confidence interval of pounds sterling 2,592 to pounds sterling 10,690 [-$4,095 to $16,890]). A one unit improvement in satisfaction was associated with extra costs in the ACT group of pounds sterling 473 ($747). CONCLUSIONS: The costs of ACT were not significantly different from usual care. ACT did, however, result in greater levels of client satisfaction and engagement with services and as such may be the preferred community treatment option for patients with long-term serious mental health problems. PMID- 19564221 TI - VA intensive mental health case management in urban and rural areas: veteran characteristics and service delivery. AB - OBJECTIVE: The availability of mental health services in rural areas- particularly intensive services such as assertive community treatment (ACT)--has been of increasing concern and was the focus of this study. In recent decades the U.S. Department of Veterans Affairs (VA) has developed a national network of ACT like programs called mental health intensive case management (MHICM), which have served veterans from diverse locations across the country, including urban and rural areas. METHODS: This study used rural-urban commuting area codes and national VA administrative data to compare characteristics of veterans and patterns of MHICM service delivery among veterans with mental illness living in large urban, large rural, small rural, and isolated rural communities. RESULTS: Among veterans enrolled in MHICM from FY 2000 to FY 2005 (N=5,221), 84% (N=4,373) resided in urban areas, 8% (N=421) in large cities, 6% (N=291) in small rural towns, and 3% (N=136) in isolated rural areas. MHICM participants who lived in rural areas had clinical problems broadly similar to those in urban areas, although more rural veterans were unemployed, disabled, received VA disability compensation, and had a payee or fiduciary. MHICM clients in smaller or isolated rural areas received slightly less frequent and less intensive contacts and less recovery-oriented services than those in large urban locations. CONCLUSIONS: These data highlight the need for intensive case management services in rural areas and note some challenges in providing them at the intensity and frequency observed in urban areas where travel distances and times are shorter. PMID- 19564219 TI - Dropout from outpatient mental health care in the United States. AB - OBJECTIVE: Although mental health treatment dropout is common, patterns and predictors of dropout are poorly understood. This study explored patterns and predictors of mental health treatment dropout in a nationally representative sample. METHODS: Data were from the National Comorbidity Survey Replication, a nationally representative household survey. Respondents who had received mental health treatment in the 12 months before the interview (N=1,664) were asked about dropout, which was defined as quitting treatment before the provider wanted them to stop. Cross-tabulation and discrete-time survival analyses were used to identify predictors. RESULTS: Approximately one-fifth (22%) of patients quit treatment prematurely. The highest dropout rate was from treatment received in the general medical sector (32%), and the lowest was from treatment received by psychiatrists (15%). Dropout rates were intermediate from treatment in the human services sector (20%) and among patients seen by nonpsychiatrist mental health professionals (19%). Over 70% of all dropout occurred after the first or second visits. Mental health insurance was associated with low odds of dropout (odds ratio=.6, 95% confidence interval=.4-.9). Psychiatric comorbidity was associated with a trend toward dropout. Several patient characteristics differentially predicted dropout across treatment sectors and in early and later phases of treatment. CONCLUSIONS: Roughly one-fifth of adults in mental health treatment dropped out before completing the recommended course of treatment. Dropout was most common in the general medical sector and varied by patient characteristics across treatment sectors. Interventions focused on high-risk patients and sectors that have higher dropout rates will likely be required to reduce the large proportion of patients who prematurely terminate treatment. PMID- 19564222 TI - Scope of agency control: assertive community treatment teams' supervision of consumers. AB - OBJECTIVE: Assertive community treatment teams have been criticized as being inherently coercive; however, base rates of control practices used by teams have not been well documented. The purpose of this study was to assess the rates at which different forms of agency control, such as involuntary outpatient commitment, representative payeeship, intensive medication monitoring, and agency supervised housing, were used by assertive community treatment teams. Also examined were program, practitioner, and consumer correlates of agency control practices. METHODS: A statewide survey was conducted of 23 assertive community treatment teams serving consumers with severe mental illness. Data were collected on both team attributes and practitioner attributes. RESULTS: Extent of agency control was highly variable across sites. Intensive medication monitoring and representative payeeship were the most frequently used agency control practices. The strongest predictor of agency control was having a higher percentage of consumers on the caseload who were diagnosed as having a schizophrenia-spectrum disorder. Fidelity to the assertive community treatment model was not associated with agency control. However, lower quality of basic clinical services (for example, assessment and treatment planning) was associated with greater use of agency-supervised housing. Pessimistic practitioner attitudes were not significantly associated with agency control, but practitioner education was negatively associated with both representative payeeship and intensive medication monitoring. CONCLUSIONS: Assertive community treatment teams differed widely in their scope of agency control, and this variation was not associated with fidelity to the model. Consumer characteristics, such as a schizophrenia spectrum disorder and active substance use, were most closely associated with agency control. PMID- 19564223 TI - Prescriber fidelity to a medication management evidence-based practice in the treatment of schizophrenia. AB - OBJECTIVE: Medication Management Approaches in Psychiatry (MedMAP) is an evidence based practice developed to guide the management and monitoring of psychotropic medications for individuals with schizophrenia. This article reports prescriber fidelity to MedMAP principles in a public mental health service system. METHODS: This three-year longitudinal intervention study implemented MedMAP in six community mental health centers in Kentucky. Nine psychiatrists and five advanced practice psychiatric nurses with prescriptive authority participated in the study. Prescribers were trained in the use of MedMAP about one month before implementation, and MedMAP principles were reinforced throughout the study. Four posttraining assessments were conducted at each site at approximately four-month intervals over a period of 30 months. An 18-item scale was used to assess baseline and posttraining prescriber fidelity over a period of 30 months in 900 randomly selected medical records. RESULTS: Average fidelity scores improved significantly over baseline at each of the four postimplementation fidelity assessments. Training effects were most evident at the second posttraining fidelity assessment, but effects persisted over the course of the study. There was considerable variability in scores across items both at baseline and subsequently. Posttraining improvement was greatest in patient education, documentation of illness and medication history, and simplification of medication regimen. CONCLUSIONS: Implementation and monitoring of MedMAP were feasible in these community mental health settings. Additional implementation projects are crucial for advancing evidence-based practice in clinical settings. PMID- 19564224 TI - Evaluation of an outpatient intervention for women with severe depression and a history of childhood trauma. AB - OBJECTIVE: The study examined the effectiveness of a three-month structured outpatient intervention developed for women with severe depression and childhood trauma that used brief psychodynamic psychotherapy by comparing it to standard treatment recommended in clinical guidelines issued by the Chilean Ministry of Health. METHODS: Eighty-seven women who sought treatment from a public health service in Curico, Chile, and who had severe depression and a history of childhood traumatic experiences were randomly assigned to receive either the intervention (N=44) or standard treatment (N=43). The participants were assessed at baseline and at three months (completion of the intervention) and six months with use of the Hamilton Depression Scale (Ham-D); Lambert's Outcome Questionnaire (OQ-45.2), which rates psychiatric symptoms, interpersonal relationships, and social role functioning; and the Post-traumatic Stress Treatment Outcome scale (PTO 8), which assesses symptoms of posttraumatic stress disorder. An intent-to-treat design was used with multiple analyses of variance. RESULTS: At three months significant differences were found in favor of the intervention group in Ham-D scores (p<.001) and OQ-45.2 scores (p<.05). At six months a significantly greater proportion of the intervention group had indicators of remission as measured by the OQ-45.2 (39% versus 14%, p<.05) and by the Ham-D (22% versus 5%, p<.05). Scores on the PTO 8 indicated significant improvements in posttraumatic stress disorder symptoms at six months, although there were no significant differences between the groups. CONCLUSIONS: An outpatient intervention that screened for and focused on childhood traumas and that helped patients understand current psychosocial difficulties as a repetition of past traumas was effective in reducing psychiatric symptoms and improving interpersonal relationships and social role functioning among women with severe depression and a history of childhood trauma. PMID- 19564225 TI - Help seeking and perceived need for mental health care among individuals in Canada with suicidal behaviors. AB - OBJECTIVE: This study examined and compared help seeking, perceived need, satisfaction with health professionals, and barriers to care in three groups: individuals with a mental disorder without suicidal behaviors, those with suicidal ideation with or without a mental disorder, and those with a suicide attempt with or without a mental disorder in the past year. METHODS: Data came from the Canadian Community Health Survey Cycle 1.2. The sample consisted of 36,984 persons aged 15 years and older (response rate=77%). A total of 4,872 had a mental disorder without suicidal behaviors, 1,234 had suicidal ideation, and 230 had attempted suicide. Multiple logistic regressions were used to examine differences between the three groups after adjusting for sociodemographic factors and the number of mental disorders. RESULTS: Individuals with suicidal ideation and those with suicide attempts were significantly more likely than those with a mental disorder but no suicidal behaviors to seek help and to perceive a need for care in the past year. However, 48% of individuals reporting suicidal ideation and 24% of individuals reporting a suicide attempt did not seek help and did not perceive a need for help in the past year. Significant differences existed between individuals in the three groups in terms of satisfaction with the care they received and barriers to receiving care in the past year. CONCLUSIONS: Although suicidal ideation and suicide attempts represent a significant source of evaluated need associated with help seeking and perceived need over and above the presence and severity of mental disorders, a significant proportion of individuals with suicidal behaviors did not receive care and did not perceive a need for care. Future research should be directed toward finding better ways to identify these individuals and address barriers to their care and other factors that may interfere with their receiving help. PMID- 19564226 TI - Children's beliefs about causes of childhood depression and ADHD: a study of stigmatization. AB - OBJECTIVE: Children's causal attributions about childhood mental health problems were examined in a national sample for prevalence; relative stigmatization; variation by age, race and ethnicity, and gender; and self-report of a diagnosis of depression or attention-deficit hyperactivity disorder (ADHD). METHODS: A national sample of 1,091 children were randomly assigned to read vignettes about a peer with depression, ADHD, or asthma and respond to an online survey. Causal attributions and social distance were assessed, and correlations were examined. Logistic regression models for each causal item tested main effects and interaction terms for conditions, demographic characteristics, and self-reported diagnosis. RESULTS: The beliefs that parenting, substance abuse, and low effort caused the condition were all strongly intercorrelated and were moderately correlated with social distance. The depression condition was the strongest predictor of endorsement of the most stigmatizing causal beliefs. Stigmatizing causal beliefs were evident for ADHD, but with more modest effects. Children who reported a diagnosis were more likely to endorse parenting and substance abuse as causes (attenuated for ADHD). Modest to moderate effects were found for variation in causal beliefs across ethnic groups. CONCLUSIONS: This study demonstrated a consistent presence of stigmatization in children's beliefs about the causes of childhood mental health problems. Low effort, parenting, and substance abuse together tapped a moralistic and blaming view of mental health problems. The results reinforce the need to address stigmatization of mental disorders and the relative stigmatization of different causal beliefs. The findings of variation by ethnicity and diagnosis can inform and target antistigmatization efforts. PMID- 19564227 TI - How many nursing home residents live with a mental illness? AB - OBJECTIVE: A number of data sets can be used to estimate the size of the nursing home population that has mental illness; however, estimates vary because of differences in methods of data collection. This study sought to compare estimates from three nationally representative data sets of the number of nursing home residents who have a mental illness, determine which data set provides the best national-level estimate, and identify the types of policy and monitoring questions that can best be answered with each. METHODS: The study compared estimates of the number of nursing home residents who had either a primary or any diagnosed mental illness from the National Nursing Home Survey (NNHS), the Minimum Data Set (MDS), and the Medicaid Analytic eXtract (MAX) files. RESULTS: The NNHS produced the most valid national-level estimates of residents with a mental illness--nearly 102,000 with a primary diagnosis in 2004 (6.8% of residents), of which about 23,000 were under age 65 and 79,000 were aged 65 and older. However, data from the NNHS cannot be broken down to the state level; therefore, state- and facility-level estimates would have to be generated with the MDS or MAX data sets. CONCLUSIONS: Policy makers and program managers need to be aware of the strengths and limitations of the data they use in order to make informed decisions. Users of the NNHS, MDS, and MAX data sets should be aware of the differences in recorded diagnoses among the three, especially the relatively limited diagnoses in the MAX and imprecise diagnoses in the MDS. PMID- 19564229 TI - Improving psychiatric services through mystery shopping. AB - Pseudo-patients, or "mystery shoppers," have been widely used in the medical setting to detect and correct deficiencies in the delivery of health care services. Persons pretending to be patients have found discrepancies between the service promised and the service delivered, leading to positive changes, such as reduced waiting times, increased retention of patients within medical practices, better explanations of procedures and proposed treatments, and improved encounters with office staff, physicians, and other health care providers. Mystery shopping in the field of mental health, however, remains an untapped strategy to improve service delivery, especially in public-sector programs. Competition among health care providers will likely encourage patients to comparison-shop for treatment, and psychiatric facilities should consider mystery shopping as an innovative method to improve the mental health care consumer's experience. PMID- 19564228 TI - Trends in mental health admissions to nursing homes, 1999-2005. AB - OBJECTIVE: The study examined 1999-2005 data on first-time nursing home admissions of individuals with mental illness, dementia, or both to identify trends and characteristics. METHODS: The Minimum Data Set was used to estimate the number and percentage of persons newly admitted to nursing homes who had mental illness (schizophrenia, bipolar disorder, depression, or an anxiety disorder), dementia, or both from 1999 to 2005. Data from 2005 were used to compare demographic characteristics and comorbid conditions of the three groups and treatments received. RESULTS: The number of individuals admitted with mental illness increased from 168,721 in 1999 to 187,478 in 2005. The 2005 number is more than 50% higher than the number admitted with dementia only (118,290 in 2005). The increase was driven by growth in admissions of persons with depression -from 128,566 to 154,262 in 2005. Persons admitted with depression had higher rates of comorbid conditions than those admitted with dementia or with neither dementia nor mental illness. They also had high rates of antidepressant treatment and high rates of receipt of training in skills required to return to the community. CONCLUSIONS: Current trends show that the proportion of nursing home admissions with mental illness, in particular depression, has overtaken the proportion with dementia. These changes may be related to increased recognition of depression, availability of alternatives to nursing homes for persons with dementia, and increased specialization among nursing homes in the care of postacute, rehabilitation residents. In light of these trends, it is critical to ensure that nursing homes have resources to adequately treat residents with mental illness to facilitate community reintegration. PMID- 19564230 TI - Revised cost estimates of Medicaid recipients with serious mental illness and HIV AIDS. AB - OBJECTIVE: The aim of this brief report is to provide accurate 1996 baseline cost estimates for persons with co-occurring HIV and serious mental illness by revising original figures previously reported by the authors. METHODS: Data were examined for 23,729 adults who were enrolled in Medicaid in 1996. A comparison of utilization and cost of services was done for four groups: serious mental illness and HIV-AIDS, serious mental illness only, HIV-AIDS only, and neither condition. RESULTS: In 1996 persons with both illnesses had the highest annual medical and behavioral health treatment expenditures at $20,038 per person, followed by persons with only HIV-AIDS at $14,714. The cost of care for the HIV-AIDS population, regardless of the presence of serious mental illness, averaged $16,253 per person. CONCLUSIONS: Although the absolute costs found in this study were much higher than those reported in the authors' previous study, both studies showed that those with co-occurring illnesses had the highest cost. PMID- 19564231 TI - Health and social characteristics of homeless adults in Manhattan who were chronically or not chronically unsheltered. AB - OBJECTIVE: This study compared health and social characteristics of two groups of homeless adults in Manhattan--those who were chronically unsheltered and those who were not. METHODS: Outreach workers conducted brief, structured interviews with 1,093 unsheltered homeless adults. Respondents were later categorized as being chronically unsheltered on the basis of New York City criteria (sleeping without shelter at least nine of the previous 24 months). RESULTS: The sample had high rates of substance abuse (65%), serious medical issues (42%), and repeated trauma (51%) and low rates of medical insurance (47%) and income entitlements (26%) entitlements. Sixty-seven percent were chronically unsheltered, and these respondents had significantly higher rates on several measures, including military service, incarceration, and mental illness. CONCLUSIONS: The sick and aged nature of this population suggests that more aggressive efforts are needed to enroll unsheltered homeless people in income and health benefits and to create adequate housing opportunities with appropriate support services. PMID- 19564232 TI - Aging fathers of adult children with schizophrenia: the toll of caregiving on their mental and physical health. AB - OBJECTIVE: The primary purpose of this study was to compare the physical and psychological well-being of caregiving fathers of an adult son or daughter with schizophrenia with a comparison group of fathers who do not have children with schizophrenia. METHODS: Data for the caregiving fathers (N=95) were drawn from a study of families of adults with schizophrenia; the comparison group of fathers (N=95) was drawn from the Wisconsin Longitudinal Study (WLS). Samples were matched on sociodemographic variables and analyzed with independent-samples t tests. RESULTS: Fathers of adults with schizophrenia experienced higher levels of depression, poorer perceived health, lower levels of psychological well-being, and less marital satisfaction compared with their age-matched peers in the WLS. CONCLUSIONS: This study found hidden personal costs of caregiving among fathers of adults with schizophrenia; strategies are needed for supporting these fathers. PMID- 19564233 TI - Burnout among psychiatrists in Milan: a multicenter survey. AB - OBJECTIVES: This study evaluated the prevalence of job burnout and estimated psychiatric morbidity and job satisfaction among psychiatrists in Milan. Also investigated were the contributions of personal and environmental factors to burnout. METHODS: Data were gathered via a cross-sectional, descriptive, multicenter survey. All psychiatrists working in departments of psychiatry within the Italian Public Health System in Milan were invited to participate, and an overall response rate of 70% (N=81) was achieved. The Maslach Burnout Inventory, the 12-item General Health Questionnaire, a job satisfaction measure, and a study specific questionnaire were used in the assessments. RESULTS: Psychiatrists showed high levels of emotional exhaustion and depersonalization. Main sources of stress were related to work environment. According to regression models, the variable that most predicted burnout was a low level of job satisfaction. CONCLUSIONS: Data suggested that psychiatrists had higher levels of burnout than other physicians employed in general medical settings and confirmed that job satisfaction could have a protective role. PMID- 19564234 TI - Understanding community mental health administrators' perspectives on dialectical behavior therapy implementation. AB - In this study, key informant interviews were conducted with 13 administrators from nine community-based mental health agencies implementing dialectical behavior therapy in order to assess their perspectives on implementation. Four major themes were identified. They include opinions about dialectical behavior therapy and its fit with existing practices, resource concerns (for example, reimbursement issues, time commitment, and staff training), staff selection for training and staff turnover, and ongoing client referrals. Understanding agency administrators' unique perspectives and addressing their concerns is critical to treatment implementation given administrators' role in determining whether and how a treatment will be implemented. Better understanding of the fit between dialectical behavior therapy and existing service structures, the impact of staff turnover on implementation, and the resources required for implementation are all needed to ensure successful implementation and sustainability. PMID- 19564236 TI - The need for a "mental health home". PMID- 19564237 TI - A useful question. PMID- 19564238 TI - Documentation of suicide risk assessment in clinical records. PMID- 19564239 TI - Biophysical annotation and representation of CellML models. AB - MOTIVATION: CellML is an implementation-independent model description language for specifying and exchanging biological processes. The focus of CellML is the representation of mathematical formulations of biological processes. The language captures the mathematical and model building constructs well, but does not lend itself to capturing the biology these models represent. RESULTS: This article describes the development of an ontological framework for annotating CellML models with biophysical concepts. We demonstrate that, by using these ontological mappings, in combination with a set of graph reduction rules, it is possible to represent the underlying biological process described in a CellML model. PMID- 19564240 TI - Correlates of intensive alcohol and drug use in men who have sex with men in Catalonia, Spain. AB - BACKGROUND: The objectives of the study were to determine the prevalence of alcohol and drug use before or during sex among men who have sex with men (MSM) in Catalonia during 2006, and to identify factors associated with variables of intensive alcohol and drug use. METHODS: Cross-sectional study using self administered questionnaires. Men were recruited in saunas, sex shops, bars and a public park and by mail to all the members of the Catalonia Gay Federation. RESULTS: 19.6% of men said they were frequent users of alcohol, some type of drug (21.7%), or that they were multidrug users (18%) in the last 12 months. The multivariate analysis showed an association between having suffered discrimination and frequent alcohol and multidrug use. Being human immunodeficiency virus (HIV)-positive was associated with frequent use of drugs and multidrug use. Associations between substance use and sexual risk behaviour also emerged. CONCLUSION: The high percentage of MSM who use alcohol and drugs before and during sex and association between these substances and sexual risk behaviours reveals the need to intensify interventions to reduce their levels of use and/or to reduce the associated damage and risks. These programs must try to cover MSM-specific psychosocial aspects and include prevention for HIV-positive men. PMID- 19564241 TI - When I use a word ... declarative titles. PMID- 19564243 TI - Brief report: development of the inflammatory bowel disease family responsibility questionnaire. AB - OBJECTIVE: To present psychometric data on youth and parent versions of the Inflammatory Bowel Disease-Family Responsibility Questionnaire (IBD-FRQ), a measure of family involvement in IBD management. METHODS: Fifty-eight adolescents with inflammatory bowel disease (IBD), along with 55 mothers and 26 fathers completed the IBD-FRQ, a demographics questionnaire, and a measure of family involvement in decision making in non-IBD domains. Medical information was obtained via chart review. RESULTS: Support for the internal consistency of the IBD-FRQ was obtained. Evidence of validity was documented via moderate to high intercorrelations among reporters. Youth involvement increased with youth age, while maternal and paternal involvement decreased with youth age. Across all reporters, maternal involvement was higher than paternal involvement. CONCLUSIONS: Preliminary analyses offer support for the measure's reliability and validity. The measure shows promise as a means of assessing family involvement in IBD condition management; however, further validation studies are needed. PMID- 19564244 TI - Prostate cancer: current evidence weighs against population screening. PMID- 19564245 TI - Screening for prostate cancer. AB - In the United States, prostate cancer will affect 1 man in 6 during his lifetime. Since the mid-1980s, screening with the prostate-specific antigen (PSA) blood test has more than doubled the risk of a prostate cancer diagnosis. A decrease in prostate cancer death rates has been observed since that time, but the relative contribution of PSA testing as opposed to other factors, such as improved treatment, has been uncertain. The recent release of 2 large randomized trials suggests that if there is a benefit of screening, it is, at best, small. Methods to assess a man's risk of prostate cancer, including those tools that integrate multiple risk factors, are now available and should be used in risk assessment. Men undergoing screening for prostate cancer may reduce their risk of prostate cancer with finasteride. PMID- 19564246 TI - Resting heart rate and blood pressure, independent of each other, proportionally raise the risk for type-2 diabetes mellitus. AB - BACKGROUND: Fast heart rate and high blood pressure (BP) at rest may raise risk for the development of type-2 diabetes mellitus (DM). We therefore investigated dose-response and interactive effects of resting heart rate and BP on the incidence of DM in a Japanese population. METHODS: A follow-up study was conducted for 16 828 men and 8368 women aged 30-59 years and apparently healthy at baseline. Incident DM was identified by 'fasting serum glucose > or =7.00 mmol/l (126 mg/dl)' or/and 'under medical treatment for DM'. Using Cox proportional hazard models, hazard ratio (HR) for incident DM were estimated according to the quartiles of heart rate, systolic or diastolic BP (SBP, DBP), and its linear trends were checked by computing the three indices as continuous variables. Subsequently, interactive effects of slow/fast heart rate (dichotomized by the median) and low/high SBP or DBP (dichotomized by the median) on HR were examined. Baseline age, body mass index, smoking, drinking, exercise and education were computed as conventional confounders. RESULTS: During the follow-up of 125 106 person-years for men and 59 616 person-years for women, 869 men and 224 women developed DM. The multivariate-adjusted HR for incident DM increased across quartiles of heart rate, SBP and DBP in both sexes (linear trend P<0.001 for all). Neither sex showed any significant interactive effects of heart rate and SBP or DBP on HR. CONCLUSIONS: Resting heart rate and BP proportionally raise the risk for DM in middle-aged healthy men and women. Moreover, the adverse effects of fast heart rate and high BP are independent of each other as well as of the influences of conventional confounders. PMID- 19564247 TI - Commentary: Trends in indigenous inequalities in mortality in New Zealand. PMID- 19564248 TI - Predicting ambient ultraviolet from routine meteorological data; its potential use as an instrumental variable for vitamin D status in pregnancy in a longitudinal birth cohort in the UK. AB - BACKGROUND: Maternal vitamin D status in pregnancy has been postulated to have important effects on intrauterine development. UVB radiation is not commonly measured but is the prime determinant of circulating 25-hydroxyvitamin-D [25 (OH)D] and is highly dependent on regional weather including cloud cover, ozone and sunshine hours. METHODS: Using linear regression we described the relationship between estimated ambient-erythemal ultraviolet (eUV) exposure in Oxford (1990-95) and total hours of sunshine and month in order to forecast eUV in nearby regions, whilst adjusting for regional variations in weather. The forecast was validated with empirical data collected from Cornwall and then predicted for the Avon region. Total 98-day prenatal ambient-eUV was then predicted in 355 expectant mothers in the Avon Longitudinal Study of Parents and Children (ALSPAC) cohort and its relationship with maternal vitamin D status was determined. RESULTS: Estimated ambient-eUV was strongly associated with measured ambient-eUV (r(2) = 0.989) with a near 1:1 prediction for the validation data set [beta = 0.99, 95% confidence interval (CI) 0.913, 1.067 r(2) = 0.980]; strong seasonal associations were observed between eUV in the last trimester of pregnancy and maternal serum 25-(OH)D concentrations (r(2) = 0.40). CONCLUSION: This technique of prediction could be applied to existing cohorts allowing the relationship between maternal vitamin D status and the health of the offspring to be studied via instrumental variable analysis. PMID- 19564249 TI - Gq-initiated cardiomyocyte hypertrophy is mediated by phospholipase Cbeta1b. AB - Activation of the heterotrimeric G protein Gq causes cardiomyocyte hypertrophy in vivo and in cell culture models. Hypertrophic responses induced by pressure or volume overload are exacerbated by increased Gq activity and ameliorated by Gq inhibition. Gq activates phospholipase Cbeta (PLCbeta) subtypes, resulting in generation of the intracellular messengers inositol(1,4,5)tris-phosphate [Ins(1,4,5)P(3)] and sn-1,2-diacylglycerol (DAG), which regulate intracellular Ca(2+) and conventional protein kinase C subtypes, respectively. Gq can also signal independently of PLCbeta, and the involvement of either Ins(1,4,5)P(3) or DAG in cardiomyocyte hypertrophy has not been unequivocally established. Overexpression of one splice variant of PLCbeta1, specifically PLCbeta1b, in neonatal rat cardiomyocytes causes increased cell size, elevated protein/DNA ratio, and heightened expression of the hypertrophy-related marker gene, atrial natriuretic peptide. The other splice variant, PLCbeta1a, had no effect. Expression of a 32-aa C-terminal PLCbeta1b peptide, which competes with PLCbeta1b for sarcolemmal association, prevented PLC activation and eliminated hypertrophic responses initiated by Gq or Gq-coupled alpha(1)-adrenergic receptors. In contrast, a PLCbeta1a C-terminal peptide altered neither PLC activity nor cellular hypertrophy. We conclude that hypertrophic responses initiated by Gq are mediated specifically by PLCbeta1b. Preventing PLCbeta1b association with the sarcolemma may provide a useful therapeutic target to limit hypertrophy. PMID- 19564250 TI - Ae2(a,b)-deficient mice exhibit osteopetrosis of long bones but not of calvaria. AB - Extracellular acidification by osteoclasts is essential to bone resorption. During proton pumping, intracellular pH (pH(i)) is thought to be kept at a near neutral level by chloride/bicarbonate exchange. Here we show that the Na(+) independent chloride/bicarbonate anion exchanger 2 (Ae2) is relevant for this process in the osteoclasts from the long bones of Ae2(a,b)(-/-) mice (deficient in the main isoforms Ae2a, Ae2b(1), and Ae2b(2)). Although the long bones of these mice had normal numbers of multinucleated osteoclasts, these cells lacked a ruffled border and displayed impaired bone resorption activity, resulting in an osteopetrotic phenotype of long bones. Moreover, in vitro osteoclastogenesis assays using long-bone marrow cells from Ae2(a,b)(-/-) mice suggested a role for Ae2 in osteoclast formation, as fusion of preosteoclasts for the generation of active multinucleated osteoclasts was found to be slightly delayed. In contrast to the abnormalities observed in the long bones, the skull of Ae2(a,b)(-/-) mice showed no alterations, indicating that calvaria osteoclasts may display normal resorptive activity. Microfluorimetric analysis of osteoclasts from normal mice showed that, in addition to Ae2 activity, calvaria osteoclasts--but not long-bone osteoclasts--possess a sodium-dependent bicarbonate transporting activity. Possibly, this might compensate for the absence of Ae2 in calvaria osteoclasts of Ae2(a,b)(-/-) mice. PMID- 19564251 TI - The CHEER study to reduce BMI in Elementary School students: a school-based, parent-directed study in Framingham, Massachusetts. AB - Childhood obesity may be lessened by parent-focused interventions. A pilot parent directed trial with 46 parents of overweight and obese elementary school students was conducted at two ethnically diverse public schools in Framingham, Massachusetts. Parents were randomly assigned to either the Materials Group, which received mailed educational materials, or the Materials plus Personal Encounters Group, which received educational materials through interactions with community health workers (CHWs). Parents completed baseline and post-intervention surveys; children's body mass index (BMI) percentiles were measured at baseline and post-intervention. There were no differences in the reduction of children's BMI between groups. However, the mean BMI percentile for all children dropped from 94.1 to 90.6 (p = .005), while there was no change in BMI among a nonrandomized contemporaneous control group. Findings are limited by the lack of a true control group and small sample size. Results from this school nurse and CHW outreach program to parents are encouraging. PMID- 19564252 TI - Peer review in diagnostic radiology: current state and a vision for the future. AB - Over the past decade, the level of interest in improving the quality of healthcare in the United States has increased. New requirements established by regulatory organizations require the ongoing practice-based evaluation of physician performance. Peer review, a key process in physician performance evaluation, is geared primarily toward measuring diagnostic accuracy. Accuracy may be measured in terms of interpretive agreement or disagreement during a blinded double reading or in workstation-integrated evaluations. Each method of assessing diagnostic accuracy has strengths and weaknesses that should be carefully considered before it is implemented in a particular departmental or institutional setting. PMID- 19564253 TI - Informatics in radiology: Render: an online searchable radiology study repository. AB - Radiology departments are a rich source of information in the form of digital radiology reports and images obtained in patients with a wide spectrum of clinical conditions. A free text radiology report and image search application known as Render was created to allow users to find pertinent cases for a variety of purposes. Render is a radiology report and image repository that pools researchable information derived from multiple systems in near real time with use of (a) Health Level 7 links for radiology information system data, (b) periodic file transfers from the picture archiving and communication system, and (c) the results of natural language processing (NLP) analysis. Users can perform more structured and detailed searches with this application by combining different imaging and patient characteristics such as examination number; patient age, gender, and medical record number; and imaging modality. Use of NLP analysis allows a more effective search for reports with positive findings, resulting in the retrieval of more cases and terms having greater relevance. From the retrieved results, users can save images, bookmark examinations, and navigate to an external search engine such as Google. Render has applications in the fields of radiology education, research, and clinical decision support. PMID- 19564254 TI - Continued impact of pneumococcal conjugate vaccine on carriage in young children. AB - OBJECTIVES: The goals were to assess serial changes in Streptococcus pneumoniae serotypes and antibiotic resistance in young children and to evaluate whether risk factors for carriage have been altered by heptavalent pneumococcal conjugate vaccine (PCV7). METHODS: Nasopharyngeal specimens and questionnaire/medical record data were obtained from children 3 months to <7 years of age in primary care practices in 16 Massachusetts communities during the winter seasons of 2000 2001 and 2003-2004 and in 8 communities in 2006-2007. Antimicrobial susceptibility testing and serotyping were performed with S pneumoniae isolates. RESULTS: We collected 678, 988, and 972 specimens during the sampling periods in 2000-2001, 2003-2004, and 2006-2007, respectively. Carriage of non-PCV7 serotypes increased from 15% to 19% and 29% (P < .001), with vaccine serotypes decreasing to 3% of carried serotypes in 2006-2007. The relative contribution of several non PCV7 serotypes, including 19A, 35B, and 23A, increased across sampling periods. By 2007, commonly carried serotypes included 19A (16%), 6A (12%), 15B/C (11%), 35B (9%), and 11A (8%), and high-prevalence serotypes seemed to have greater proportions of penicillin nonsusceptibility. In multivariate models, common predictors of pneumococcal carriage, such as child care attendance, upper respiratory tract infection, and the presence of young siblings, persisted. CONCLUSIONS: The virtual disappearance of vaccine serotypes in S pneumoniae carriage has occurred in young children, with rapid replacement with penicillin nonsusceptible nonvaccine serotypes, particularly 19A and 35B. Except for the age group at highest risk, previous predictors of carriage, such as child care attendance and the presence of young siblings, have not been changed by the vaccine. PMID- 19564255 TI - State and regional variation in regulations related to feeding infants in child care. AB - OBJECTIVE: The purpose of this study was to compare state and regional variation in infant feeding regulations for child care facilities and to compare these regulations to national standards. METHODS: We reviewed regulations for child care for all US states and Washington, DC, and examined patterns according to type of facility and geographic region. We compared state regulations with national standards for feeding infants in child care. The standards included were: (1) infants are fed according to a feeding plan from a parent or physician; (2) breastfeeding is supported by the child care facility; (3) no solid food is given before 6 months of age; (4) infants are fed on demand; (5) infants are fed by a consistent caregiver; (6) infants are held while feeding; (7) infants cannot carry or sleep with a bottle; (8) caregivers cannot feed >1 infant at a time; (9) no cow's milk is given to children <12 months of age; (10) whole cow's milk is required for children 12 to 24 months of age; and (11) no solid food is fed in a bottle. RESULTS: The mean number of regulations for states was 2.8 (SD: 1.6) for centers and 2.0 (SD: 1.3) for family child care homes. No state had regulations for all 11 standards for centers; only Delaware had regulations for 10 of the 11 standards. For family child care homes, Ohio had regulations for 5 of the 11 standards, the most of any state. States in the South had the greatest mean number of regulations for centers (3.3) and family child care homes (2.2), and the West had the fewest (2.3 and 1.9, respectively). CONCLUSIONS: Many states lacked infant feeding regulations. Encouraging states to meet best-practice national standards helps ensure that all child care facilities engage in appropriate and healthful infant feeding practices. PMID- 19564256 TI - Conflicts about end-of-life decisions in NICUs in the Netherlands. AB - OBJECTIVE: To determine the frequency and background of conflicts about neonatal end-of-life (EoL) decisions. METHODS: We reviewed the medical files of 359 newborns who had died during 1 year in the 10 Dutch NICUs and identified 150 deaths that were preceded by an EoL decision on the basis of the child's poor prognosis. The attending neonatologists of 147 of the 150 newborns were interviewed to obtain details about the decision-making process. RESULTS: EoL decisions about infants with a poor prognosis were initiated mainly by the physician, who subsequently involved the parents. Conflicts between parents and the medical team occurred in 18 of 147 cases and were mostly about the child's poor neurologic prognosis. Conflicts within the team occurred in 6 of 147 cases and concerned the uncertainty of the prognosis. In the event of conflict, the EoL decision was postponed. Consensus was reached by calling additional meetings, performing additional diagnostic tests, or obtaining a second opinion. The chief causes of conflict encountered by the physicians were religious convictions that forbade withdrawal of life-sustaining treatment and poor communication between the parents and the team. CONCLUSIONS: The parents were involved in all EoL decision-making processes, and consensus was ultimately reached in all cases. Conflicts within the team occurred in 4% of the cases and between the team and the parents in 12% of the cases. The conflicts were resolved by postponing the EoL decision until consensus was achieved. PMID- 19564257 TI - Headache in young children in the emergency department: use of computed tomography. AB - OBJECTIVE: The goal was to determine whether computed tomographic (CT) scans led to better acute care of young children with headache presenting the emergency department (ED). METHODS: We examined the records of 364 children 2 to 5 years of age who presented with headache to a large urban ED between July 1, 2003, and June 30, 2006. By reviewing initial history and examination findings, we first identified patients with secondary headaches (ie, with readily identifiable explanations such as ventriculoperitoneal shunts, known brain tumors, or acute illnesses, such as viral syndromes, fever, probable meningitis, or trauma). Charts for the remaining patients were reviewed for headache history, neurologic examination findings, laboratory and neuroimaging results, final diagnosis, and disposition. RESULTS: On the basis of initial history and physical examination results, 306 children (84%) had secondary headaches. For 72% of those children, acute febrile illnesses and viral respiratory syndromes accounted for the headaches. Among the 58 children (16%) who had no recognized central nervous system disease or systemic illness at presentation, 28% had CT scans performed. Of those, 1 scan yielded abnormal results, showing a brainstem glioma; the patient demonstrated abnormal neurologic examination findings on the day of presentation. For 15 (94%) of 16 patients, the CT scans did not contribute to diagnosis or management. For 59% of children with apparently primary headaches, no family history was recorded. CONCLUSION: For young children presenting to the ED with headache but normal neurologic examination findings and nonworrying history, CT scans seldom lead to diagnosis or contribute to immediate management. PMID- 19564258 TI - Preventive dental care for young, Medicaid-insured children in Washington state. AB - BACKGROUND: Children from low-income families face barriers to preventive dental care (PDC) and are disproportionately affected by dental caries. The Access to the Baby and Childhood Dentistry (ABCD) program of Washington State is targeted to Medicaid-insured children <6 years of age to improve their access to PDC. OBJECTIVES: To test the hypothesis that residing in an ABCD county improves the likelihood of receiving PDC and, to compare PDC use among young, Medicaid-insured children in Washington to national statistics. METHODS: We extracted 2003 Washington Medicaid dental claims for continuously enrolled children or=1 preventive dental visit (PDV) in 2003. For national comparison, we used the 2003 Medical Expenditure Panel Survey (MEPS). RESULTS: Among Medicaid-insured children or=1 PDV in 2003. We also found that significantly more children in established ABCD counties received PDC compared with privately insured US children. These findings provide additional evidence that the ABCD program reduces disparities in dental care access among young, Medicaid-insured children in Washington and point to the importance of expanding the ABCD program to other states. PMID- 19564259 TI - Prenatal growth and early postnatal influences on adult motor cortical excitability. AB - OBJECTIVE: Suboptimal prenatal growth may adversely influence motor neurophysiologic development and predispose the individual to greater risk of neurodegenerative disorders in later life. We investigated the influences of prenatal growth and the postnatal environment on motor cortical function in young adults. METHODS: Transcranial magnetic stimulation was used to construct corticospinal stimulus-response curves for 35 young adults (mean age: 28 +/- 0.5 years; 19 males) born >or=37 weeks' gestation. Birth weight centile was calculated relative to maternal size, parity, ethnicity, gender, and gestation. Handgrip strength and dexterity were measured separately. Regression analyses assessed the influence of prenatal (birth weight centile and gestation) and postnatal (socioeconomic indices and maternal education) factors on corticospinal parameters, strength, and dexterity scores. RESULTS: Lower birth weight was associated with increased interhemispheric asymmetry in motor threshold and increased cortical stimulus-response curve slope. A shorter gestation predicted a larger area under this curve in the right hand. High motor threshold was predicted by greater environmental adversity in early postnatal life, but not by prenatal factors. Higher birth weight centile and lower motor threshold were associated with greater educational achievement. CONCLUSIONS: Poor in utero growth and mild prematurity are associated with altered corticospinal excitability in adulthood. An early postnatal environment with less early postnatal socioeconomic disadvantage and having a mother with a completed high school education partly ameliorates this. While altered cortical development has some functional consequences already evident in early adulthood, it may have a later, additional adverse impact on aging-related changes in motor function. PMID- 19564261 TI - Should a head-injured child receive a head CT scan? A systematic review of clinical prediction rules. AB - CONTEXT: Given radiation- and sedation-associated risks, there is uncertainty about which children with head trauma should receive cranial computed tomography (CT) scanning. A high-quality and high-performing clinical prediction rule may reduce this uncertainty. OBJECTIVE: To systematically review the quality and performance of published clinical prediction rules for intracranial injury in children with head injury. METHODS: Medline and Embase were searched in December 2008. Studies were selected if they included clinical prediction rules involving children aged 0 to 18 years with a history of head injury. Prediction-rule quality was assessed by using 14 previously published items. Prediction-rule performance was evaluated by rule sensitivity and the predicted frequency of CT scanning if the rule was used. RESULTS: A total of 3357 titles and abstracts were assessed, and 8 clinical prediction rules were identified. For all studies, the rule derivations were reported; no study validated a rule in a separate population or assessed its impact in actual practice. The rules differed considerably in population, predictors, outcomes, methodologic quality, and performance. Five of the rules were applicable to children of all ages and severities of trauma. Two of these were high quality (>or=11 of 14 quality items) and had high performance (lower confidence limits for sensitivity >0.95 and required or=13). One of these had high quality (11 of 14 quality items) and high performance (lower confidence limit for sensitivity = 0.94 and required 13% to undergo CT). Four of the 8 rules were applicable to young children, but none exhibited adequate quality or performance. CONCLUSIONS: Eight clinical prediction-rule derivation studies were identified. They varied considerably in population, methodologic quality, and performance. Future efforts should be directed toward validating rules with high quality and performance in other populations and deriving a high-quality, high-performance rule for young children. PMID- 19564260 TI - Intrapartum antibiotic exposure and early neonatal, morbidity, and mortality in Africa. AB - BACKGROUND: Infants born to women who receive intrapartum antibiotics may have higher rates of infectious morbidity and mortality than unexposed infants. OBJECTIVE: Our goal was to determine the association of maternal intrapartum antibiotics and early neonatal morbidity and mortality. METHODS: We performed secondary analysis of data from a multisite randomized, placebo-controlled clinical trial of antibiotics to prevent chorioamnionitis-associated mother-to child transmission of HIV-1 and preterm birth in sub-Saharan Africa. Early neonatal morbidity and mortality were analyzed. In an intention-to-treat (ITT) analysis, infants born to women randomly assigned to antibiotics or placebo were compared. In addition, non-ITT analysis was performed because some women received nonstudy antibiotics for various clinical indications. RESULTS: Overall, 2659 pregnant women were randomly assigned. Of these, 2466 HIV-1-infected and HIV-1 uninfected women delivered 2413 live born and 84 stillborn infants. In the ITT analysis, there were no significant associations between exposure to antibiotics and early neonatal outcomes. Non-ITT analyses showed more illness at birth (11.2% vs 8.6%, P = .03) and more admissions to the special care infant unit (12.6% vs 9.8%, P = .04) among infants exposed to maternal intrapartum antibiotics than among unexposed infants. Additional analyses revealed greater early neonatal morbidity and mortality among infants of mothers who received nonstudy antibiotics than of mothers who received study antibiotics. CONCLUSIONS: There is no association between intrapartum exposure to antibiotics and early neonatal morbidity or mortality. The associations observed in non-ITT analyses are most likely the result of women with peripartum illnesses being more likely to receive nonstudy antibiotics. PMID- 19564262 TI - Pathways to approval of pediatric cardiac devices in the United States: challenges and solutions. AB - Patients treated by pediatric interventional cardiologists and cardiac surgeons often have unmet medical device needs that pose a challenge to the current regulatory evaluation and approval process in the United States. In this report we review current US Food and Drug Administration regulatory processes, review some unique aspects of pediatric cardiology and cardiac surgery that pose challenges to these processes, and discuss possible alternate pathways to cardiac device evaluation and approval for children. Children deserve to benefit from new and refined cardiac devices and technology designed explicitly for their conditions. PMID- 19564263 TI - Assessing the functional status of hospitalized children. PMID- 19564264 TI - Policy statement--Equipment for ambulances. PMID- 19564266 TI - Clinical predictors of pneumonia among children with wheezing. AB - OBJECTIVE: The goal was to identify factors associated with radiographically confirmed pneumonia among children with wheezing in the emergency department (ED) setting. METHODS: A prospective cohort study was performed with children or=38 degrees C (positive LR: 2.03 [95% CI: 1.34-3.07]), maximal temperature in the ED of >or=38 degrees C (positive LR: 1.92 [95% CI: 1.48-2.49]), and triage oxygen saturation of <92% (positive LR: 3.06 [95% CI: 1.15-8.16]) were associated with increased risk of pneumonia. Among afebrile children (temperature of <38 degrees C) with wheezing, the rate of pneumonia was very low (2.2% [95% CI: 1.0-4.7]). CONCLUSIONS: Radiographic pneumonia among children with wheezing is uncommon. Historical and clinical factors may be used to determine the need for chest radiography for wheezing children. The routine use of chest radiography for children with wheezing but without fever should be discouraged. PMID- 19564265 TI - Functional Status Scale: new pediatric outcome measure. AB - OBJECTIVE: The goal was to create a functional status outcome measure for large outcome studies that is well defined, quantitative, rapid, reliable, minimally dependent on subjective assessments, and applicable to hospitalized pediatric patients across a wide range of ages and inpatient environments. METHODS: Functional Status Scale (FSS) domains of functioning included mental status, sensory functioning, communication, motor functioning, feeding, and respiratory status, categorized from normal (score = 1) to very severe dysfunction (score = 5). The Adaptive Behavior Assessment System II (ABAS II) established construct validity and calibration within domains. Seven institutions provided PICU patients within 24 hours before or after PICU discharge, high-risk non-PICU patients within 24 hours after admission, and technology-dependent children. Primary care nurses completed the ABAS II. Statistical analyses were performed. RESULTS: A total of 836 children, with a mean FSS score of 10.3 (SD: 4.4), were studied. Eighteen percent had the minimal possible FSS score of 6, 44% had FSS scores of >or=10, 14% had FSS scores of >or=15, and 6% had FSS scores of >or=20. Each FSS domain was associated with mean ABAS II scores (P < .0001). Cells in each domain were collapsed and reweighted, which improved correlations with ABAS II scores (P < .001 for improvements). Discrimination was very good for moderate and severe dysfunction (ABAS II categories) and improved with FSS weighting. Intraclass correlations of original and weighted total FSS scores were 0.95 and 0.94, respectively. CONCLUSIONS: The FSS met our objectives and is well suited for large outcome studies. PMID- 19564267 TI - The use of internet-based technology to tailor well-child care encounters. AB - OBJECTIVE: The goal was to evaluate the feasibility and acceptance of a new model for well-child care (WCC) in a large health maintenance organization. METHODS: We designed a new model of WCC that engages families in Internet-based developmental and behavioral screening, allows for review of the results before the visit, and allows for selection of the appropriate visit type (e-visit, e-visit with brief provider visit, or extended encounter). The new model was pilot-tested in 2 practices within a large health maintenance organization. Seven providers and 70 parents participated in the study. Parents and providers were surveyed regarding their experience and satisfaction with the encounter. RESULTS: Seventy-five percent of parents thought that the online previsit assessment improved or very much improved the WCC visit. However, 12% of parents found the online assessment somewhat or very difficult to use. All of the parents found the e-visit or the e visit with brief provider visit acceptable or very acceptable, compared with a standard WCC visit. All 7 providers thought that use of the new model helped focus the visit and that they would continue or definitely continue to use the model. CONCLUSIONS: We demonstrated the feasibility of a new model of WCC that engaged parents in previsit assessment and used alternative visit types to tailor care to the needs of the family. Future research will be needed to examine the impact of this model on important WCC outcomes. PMID- 19564268 TI - Premarital sexual intercourse among adolescents in an Asian country: multilevel ecological factors. AB - OBJECTIVE: The goal was to assess personal and environmental factors associated with premarital sex among adolescents. METHODS: We conducted a case-control study. Between 2006 and 2008, we recruited 500 adolescents who reported having engaged in voluntary sex for most recent sex. Five hundred control subjects were matched for age, gender, and ethnicity. RESULTS: Independently significant factors for premarital sex among boys were pornography viewing (adjusted odds ratio [OR]: 5.82 [95% confidence interval [CI]: 2.34-14.43]), lack of confidence to resist peer pressure (OR: 3.84 [95% CI: 2.27-6.50]), perception that more than one half of their friends had engaged in sex (OR: 3.37 [95% CI: 1.92-5.92]), permissiveness regarding premarital sex (OR: 3.41 [95% CI: 2.10-5.55]), involvement in gang activities (OR: 3.45 [95% CI: 1.66-7.15]), drinking (OR: 1.77 [95% CI: 1.07-2.94]), smoking (OR: 1.91 [95% CI: 1.14-3.20]), and living in low cost housing (OR: 3.25 [95% CI: 1.64-6.43]). For girls, additional factors were previous sexual abuse (OR: 7.81 [95% CI: 2.50-24.41]) and dropping out of school (OR: 2.72 [95% CI: 1.32-5.61]), and stronger associations were found for lack of confidence to resist peer pressure (OR: 5.56 [95% CI: 2.94-10.53]) and permissiveness regarding premarital sex (OR: 6.25 [95% CI: 3.30-11.83]). Exposure to persons with HIV/AIDS or sexually transmitted infections in the media was negatively associated with sex for boys (OR: 0.27 [95% CI: 0.16-0.45]) and girls (OR: 0.24 [95% CI: 0.13-0.47]). CONCLUSION: Sex education programs for adolescents must address social, media, and pornographic influences and incorporate skills to negotiate sexual abstinence. PMID- 19564269 TI - Nocturnal enuresis and overweight are associated with obstructive sleep apnea. AB - OBJECTIVE: The objective of this study was to examine the relationship of obstructive sleep apnea (OSA), body weight (BMI percentage [BMI%]), and monosymptomatic nocturnal enuresis (MNE) in children. METHODS: A case-control study design was used. All children were 5 to 15 years of age; case patients were recruited from a comprehensive sleep disorders center (n = 149), and control subjects were recruited from a general pediatric practice in the same catchment area (n = 139). Case patients were subject to overnight polysomnograms and grouped into apnea severity categories (minimal, mild, moderate, or severe) on the basis of respiratory disturbance index and minimum arterial oxygen saturation levels. Data for all children included age; gender; height; weight; and history of MNE, snoring, diabetes, nasal allergies, and/or enlarged tonsils. BMI% was used to group children into weight categories as suggested by the Centers for Disease Control and Prevention (underweight, normal weight, at risk for overweight, and overweight). Two age groupings were created (5-10 years and 11-15 years). Descriptive statistics provided the prevalence of OSA, weight category, and MNE among case patients and control subjects. Cross-tabulations examined the relationship of severity of OSA with weight categories and MNE, stratified by age and gender. A series of logistic regression models explored the interrelationship of the grouping variables. RESULTS: A large majority (79.9%) of control subjects were at risk for overweight, and a large majority (80.0%) of children with MNE also had some degree of OSA. Logistic regression demonstrated that both MNE (odds ratio: 5.29) and overweight (odds ratio 4.16) were significantly associated with OSA but not with each other. CONCLUSIONS: Overweight and MNE are associated with OSA but not with each other. OSA should be considered in overweight children with MNE, especially when they display other symptoms of OSA or fail to respond to standard MNE treatment programs. PMID- 19564270 TI - Maternal perception of weight status and health risks associated with obesity in children. AB - OBJECTIVE: To examine factors associated with the maternal perception of the weight status in related and unrelated children and to examine whether associated health risks for children's physical and mental health are recognized. PATIENTS AND METHODS: Two hundred nineteen mothers with children between 3 and 6 years of age took part in this study. The participating mothers were recruited from inpatient clinics and kindergartens. Parents were presented with 9 silhouettes representing different age- and gender-specific BMI percentiles. Demographic and weight-related variables were assessed with regard to their influence on the accuracy of the maternal weight estimation in general and for their own child. RESULTS: Of the participating mothers, 64.5% identified the overweight silhouettes of preschool-aged children correctly. However, only 48.8% of the mothers identified the overweight silhouettes associated with an increased risk for physical health problems, and 38.7% identified the silhouettes associated with an increased mental health risk. Mothers with a lower educational background were more likely to misclassify the overweight silhouettes and underestimate the associated health problems. For their own child, only 40.3% of the mothers chose silhouettes that were in agreement with the objective weight status of their child. This underestimation was associated with a higher maternal and child weight status but not with a general inability to identify the weight status of children. CONCLUSIONS: Identifying unrelated overweight silhouettes is influenced by maternal education level, whereas estimating their own child's weight status is influenced by the weight status of the mother and the child. Hence, feedback on the child's risk to become overweight is necessary to increase maternal risk awareness and willingness to take part in prevention programs. PMID- 19564271 TI - Estimation of optimal CPR chest compression depth in children by using computer tomography. AB - OBJECTIVE: Pediatric consensus-driven cardiopulmonary resuscitation guidelines target chest compression (CC) depths of one third to one half anterior-posterior (AP) chest depth. Estimates for this target as assessed by computed tomography (CT) measurements of internal and external AP chest dimensions could direct future pediatric cardiopulmonary resuscitation guidelines. METHODS: A total of 280 consecutive chest CT scans in permuted blocks of 20 for each of 14 age divisions between 0 and 8 years were reconstructed and analyzed. External and internal AP depths were measured at midsternum, and residual chest depth was calculated at simulated one-third and one-half AP compressions. RESULTS: After a simulated compression calculation, one-half external AP depth CC would result in residual internal depth of <10 mm for 94% (263 of 280) of children 3 months to 8 years. For a one-third external AP CC, only 0.4% (1 of 280) of children 3 months to 8 years had a calculated residual internal chest depth <10 mm. CONCLUSIONS: By using CT reconstruction estimates of chest dimensions across the developmental spectrum from 0 to 8 years of age, we demonstrated that a simulated CC targeting approximately one-third external AP chest depth seems radiographically appropriate for children aged 3 months to 8 years, whereas simulated CC targeting approximately one-half external AP chest depth seems radiographically to be too deep, resulting in residual internal chest depth of <10 mm for most patients of this age. PMID- 19564272 TI - Cardiac safety of methylphenidate versus amphetamine salts in the treatment of ADHD. AB - OBJECTIVES: Safety concerns about central nervous system stimulants for the treatment of attention-deficit/hyperactivity disorder (ADHD) include adverse cardiac effects. This study aimed to compare the risk for cardiac events in users of methylphenidate and amphetamine salts. METHODS: A retrospective cohort design using claims data from the Florida Medicaid fee-for-service program representing a total of 2131953 children and adolescents was used. The analysis included all beneficiaries who were between 3 and 20 years of age, enrolled between July 1994 and June 2004, had at least 1 physician diagnosis of ADHD and were newly started on methylphenidate or amphetamine salts. Each month of follow-up was classified according to stimulant use into current use or former use. We defined cardiac events as first emergency department (ED) visit for cardiac disease or symptoms. Risk between current users of methylphenidate versus amphetamine salts and former users of drugs in these categories was compared by using a time-dependent Cox proportional hazard model that adjusted for differences in gender; race; age; year of the index date; disability; congenital anomalies; history of circulatory disease; history of hospital admission; and use of antidepressants, antipsychotics, and bronchodilators. RESULTS: A total of 456 youth visited the ED for cardiac reasons during 52783 years of follow-up. After adjustment for differences in covariates, the risk for cardiac ED visits was similar among current users of methylphenidate or amphetamines. Periods of former use had a similar risk between youth with an exposure history to methylphenidate or amphetamine. CONCLUSION: Exposure to methylphenidate and amphetamines salts showed similar risk for cardiac ED visits. Additional population-based studies that address manifestation of serious heart disease, especially after long-term use, dosage comparisons, and interactions with preexisting cardiac risk factors are needed to inform psychiatric treatment decisions. PMID- 19564273 TI - Health status and behavioral outcomes for youth who anticipate a high likelihood of early death. AB - OBJECTIVE: The relationship between adolescents' perceived risk for dying and their involvement in risk behaviors is unknown. We sought to determine the proportion of US youth who anticipate a high likelihood of early mortality and relationships with health status and risk behaviors over time. METHODS: We analyzed data from times 1 (1995), 2 (1996), and 3 (2001-2002) of the National Longitudinal Study of Adolescent Health, a nationally representative sample of youth in grades 7 through 12. The relationship between perceived risk for premature mortality and health behaviors/outcomes was assessed by using bivariate and multivariate analyses. RESULTS: At time 1, 14.7% of the 20594 respondents reported at least a 50/50 chance that they would not live to age 35. In adjusted models, illicit drug use, suicide attempt, fight-related injury, police arrest, unsafe sexual activity, and a diagnosis of HIV/AIDS predicted early death perception at time 2, time 3, or both (adjusted odds ratios: 1.26-5.12). Conversely, perceived early mortality at time 1 predicted each of these behaviors and outcomes, except illicit drug use, at time 2 or time 3, most strongly a diagnosis of HIV/AIDS (adjusted odds ratios: 7.13 [95% confidence interval: 2.50 20.36]). CONCLUSIONS: Adolescent involvement in risk behaviors predicted a belief in premature mortality 1 and 7 years later. Reciprocally, adolescents' perceived risk for early death predicted serious health outcomes, notably a diagnosis of HIV/AIDS in young adulthood. Given its frequency and influence on behavior and health, adolescents' perceived risk for early death should be incorporated into psychosocial assessments and interviews. PMID- 19564274 TI - Risk factors for persistent fatigue with significant school absence in children and adolescents. AB - OBJECTIVE: To assess children and adolescents with severe fatigue who are referred to pediatricians and to examine whether factors can be identified at their first visit that predict worse outcomes at 1 year. METHODS: Ninety-one patients, aged 8 to 18 years completed questionnaires about sleep, somatic symptoms, physical activity, and fatigue. They were reassessed 12 months later. Measurements at baseline and outcome were analyzed by using univariable logistic regression with persistent, severe fatigue (yes/no) and persistent school absence (yes/no) as dependent variables and baseline scores as independent variables. RESULTS: After 12 months, 50.6% of the children and adolescents showed improvement; 29.1% had persistent fatigue, and 20.3% had persistent fatigue with significant school absence. Factors associated with the poorest outcome were sleep problems (odds ratio [OR]: 1.4 [95% confidence interval (CI): 1.1-1.8]), initial fatigue score (OR: 1.1 [95% CI: 1.0-1.2]), somatic complaints such as hot and cold spells (OR: 1.9 [95% CI: 1.2-3.0]), blurred vision (OR: 2.1 [95% CI: 1.1 4.0]), pain in arms and legs (OR: 2.0 [95% CI: 1.0-3.2]), back pain (OR: 1.8 [95% CI: 1.0-3.2]), constipation (OR: 1.7 [95% CI: 1.0-2.7]), and memory deficits (OR: 1.8 [95% CI: 1.0-3.2]). Resolved fatigue was associated with male gender (OR: 5.0 [95% CI: 1.6-15.5]) and a physically active lifestyle (OR: 1.3 [95% CI: 1.1 1.5]). CONCLUSIONS: Assessment of predictive factors at the first visit enables the pediatrician to identify those patients with severe fatigue who are at risk of a poor outcome. Female gender, poor sleep quality, physically inactive lifestyle, and specific somatic complaints were important predictive factors. PMID- 19564275 TI - Paternal depressive symptoms during pregnancy are related to excessive infant crying. AB - OBJECTIVE: Excessive infant crying, or infantile colic, is a common and often stress-inducing problem for parents that can ultimately result in child abuse. From previous research it is known that maternal depression is related to excessive crying, but so far little is known about the influence of paternal depression. METHODS: In a prospective, population-based study, we obtained information on both maternal and paternal depressive symptoms at 20 weeks of pregnancy by using the Brief Symptom Inventory. Parental depressive symptoms were related to excessive crying in 4426 two-month-old infants. The definition of excessive crying was based on the widely used Wessel's criteria (ie, crying >3 hours for >3 days in the past week). RESULTS: After adjustment for depressive symptoms of the mother and relevant confounders, we found a 1.29 (95% confidence interval: 1.09-1.52) higher risk of excessive infant crying per SD of paternal depressive symptoms. CONCLUSIONS: Our findings indicate that paternal depressive symptoms during pregnancy might be a risk factor for excessive infant crying. This finding could be related to genetic transmission, interaction of a father with lasting depressive symptoms with the infant, or related indirectly through contextual stressors such as marital, familial, or economic distress. PMID- 19564276 TI - Treatment of Kawasaki disease: analysis of 27 US pediatric hospitals from 2001 to 2006. AB - OBJECTIVES: We sought to analyze trends in admissions and to describe therapies used for acute Kawasaki disease over a 6-year period. METHODS: The Pediatric Health Information System provides patient data including demographic variables, International Classification of Diseases, Ninth Revision codes, and services billed to patients. Patient identifiers enable tracking of medication use in and across multiple admissions within a center. We analyzed data for patients with (1) a diagnosis code for Kawasaki disease, (2) intravenously administered immunoglobulin treatment during hospitalization, and (3) discharge between January 1, 2001, and December 30, 2006, from 27 hospitals contributing complete data over the study period. RESULTS: During the study period, 5197 Kawasaki disease admissions were identified for 4811 patients; numbers increased 32.6% from 2001 (n = 678) to 2006 (n = 899). Retreatment with intravenous immunoglobulin was administered to 712 patients (14.8%) over the study period. Other antiinflammatory therapies included intravenously administered methylprednisolone (5.8%), orally administered prednisone (2.8%), and infliximab (1%). Use of infliximab steadily increased from 0.0% (0 of 678 patients) in 2001 to 2.3% (21 of 899 patients) in 2006. Coronary artery aneurysms were coded for 3.3% of patients. Male patients, patients <1 year of age, and Hispanic patients were significantly more likely to have coding for coronary artery aneurysms. CONCLUSIONS: Our report provides the first large multicenter description of agents used in the treatment of intravenously administered immunoglobulin resistant Kawasaki disease in the United States. Trends include increased numbers of admissions attributable to Kawasaki disease and increased usage of infliximab. PMID- 19564278 TI - Urinary tract infections in 1- to 3-month-old infants: ambulatory treatment with intravenous antibiotics. AB - OBJECTIVE: The goal was to examine the feasibility of outpatient management for 1 to 3-month-old infants with febrile urinary tract infections. METHODS: A cohort study was performed with all children 30 to 90 days of age who were evaluated for presumed febrile urinary tract infections in the emergency department of a tertiary-care pediatric hospital between January 1, 2005, and September 30, 2007. Patients were treated with intravenously administered antibiotics as outpatients in a day treatment center unless they met exclusion criteria, in which case they were hospitalized. RESULTS: Of 118 infants included in the study, 67 (56.8%) were admitted to the day treatment center and 51 (43.2%) were hospitalized. The median age of day treatment center patients was 66 days (range: 33-85 days). The diagnosis of urinary tract infection was confirmed for 86.6% of patients treated in the day treatment center. Escherichia coli was identified in 84.5% of urine cultures; 98.3% of isolates were sensitive to gentamicin. Six blood cultures (10.3%) yielded positive results, 5 of them for E coli. Treatment with intravenously administered antibiotics in the day treatment center lasted a mean of 2.7 days. The mean number of visits, including appointments for voiding cystourethrography, was 2.9 visits. The rate of parental compliance with day treatment center visits was 98.3%. Intravenous access problems were seen in 8.6% of cases. Successful treatment in the day treatment center (defined as attendance at all visits, normalization of temperature within 48 hours, negative control urine and blood culture results, if cultures were performed, and absence of hospitalization from the day treatment center) was obtained for 86.2% of patients with confirmed urinary tract infections. CONCLUSIONS: Ambulatory treatment of infants 30 to 90 days of age with febrile urinary tract infections by using short term, intravenous antibiotic therapy at a day treatment center is feasible. PMID- 19564277 TI - Effectiveness of amoxicillin/clavulanate potassium in the treatment of acute bacterial sinusitis in children. AB - OBJECTIVE: The role of antibiotic therapy in managing acute bacterial sinusitis (ABS) in children is controversial. The purpose of this study was to determine the effectiveness of high-dose amoxicillin/potassium clavulanate in the treatment of children diagnosed with ABS. METHODS: This was a randomized, double-blind, placebo-controlled study. Children 1 to 10 years of age with a clinical presentation compatible with ABS were eligible for participation. Patients were stratified according to age (<6 or >or=6 years) and clinical severity and randomly assigned to receive either amoxicillin (90 mg/kg) with potassium clavulanate (6.4 mg/kg) or placebo. A symptom survey was performed on days 0, 1, 2, 3, 5, 7, 10, 20, and 30. Patients were examined on day 14. Children's conditions were rated as cured, improved, or failed according to scoring rules. RESULTS: Two thousand one hundred thirty-five children with respiratory complaints were screened for enrollment; 139 (6.5%) had ABS. Fifty-eight patients were enrolled, and 56 were randomly assigned. The mean age was 66 +/- 30 months. Fifty (89%) patients presented with persistent symptoms, and 6 (11%) presented with nonpersistent symptoms. In 24 (43%) children, the illness was classified as mild, whereas in the remaining 32 (57%) children it was severe. Of the 28 children who received the antibiotic, 14 (50%) were cured, 4 (14%) were improved, 4 (14%) experienced treatment failure, and 6 (21%) withdrew. Of the 28 children who received placebo, 4 (14%) were cured, 5 (18%) improved, and 19 (68%) experienced treatment failure. Children receiving the antibiotic were more likely to be cured (50% vs 14%) and less likely to have treatment failure (14% vs 68%) than children receiving the placebo. CONCLUSIONS: ABS is a common complication of viral upper respiratory infections. Amoxicillin/potassium clavulanate results in significantly more cures and fewer failures than placebo, according to parental report of time to resolution of clinical symptoms. PMID- 19564279 TI - Age-related renal parenchymal lesions in children with first febrile urinary tract infections. AB - OBJECTIVE: The aim of this study was to define the association between age and the occurrence of acute pyelonephritis and renal scars. METHODS: Between 1999 and 2002, all children 0 to 14 years of age consecutively seen with a first febrile urinary tract infection were enrolled in the study. (99m)Tc-Dimercaptosuccinic acid renal scintigraphy was performed within 5 days after admission for the detection of renal parenchymal involvement. The presence of vesicoureteral reflux was assessed by using cystography performed 1 month after the infection. If the acute scan results were abnormal, then follow-up (99m)Tc-dimercaptosuccinic acid scanning was performed after 6 months, to assess the frequency of scars. RESULTS: A total of 316 children were enrolled in the study (190 children <1 year, 99 children 1-4 years, and 27 children 5-14 years of age). (99m)Tc Dimercaptosuccinic acid scintigraphy revealed that 59% of the children had renal involvement in the acute phase of infection. The frequency of kidney involvement in infants <1 year of age (49%) was significantly lower than that in children 1 to 4 years of age (73%) and >5 years of age (81%). Of the 187 children with positive acute (99m)Tc-dimercaptosuccinic acid scan results, 123 underwent repeat scintigraphy after 6 months. Renal scars were found for 28% of children <1 year, 37% of children 1 to 4 years, and 53% of children 5 to 14 years of age. No significant differences in the frequency of scars and the presence or absence of vesicoureteral reflux were noted. CONCLUSIONS: Our findings confirm that acute pyelonephritis and subsequent renal scarring occur only in some children with first febrile urinary tract infections. Children <1 year of age with febrile urinary tract infections have a lower risk of parenchymal localization of infection and renal scarring. PMID- 19564280 TI - Influenza virus infection and the risk of serious bacterial infections in young febrile infants. AB - OBJECTIVE: We aimed to determine the risk of SBIs in febrile infants with influenza virus infections and compare this risk with that of febrile infants without influenza infections. PATIENTS AND METHODS: We conducted a multicenter, prospective, cross-sectional study during 3 consecutive influenza seasons. All febrile infants or=5 x 10(4) colony forming units per mL or >or=10(4) colony-forming units per mL in association with a positive urinalysis. Bacteremia, bacterial meningitis, and bacterial enteritis were defined by growth of a known bacterial pathogen. SBI was defined as any of the 4 above-mentioned bacterial infections. RESULTS: During the 3-year study period, 1091 infants were enrolled. A total of 844 (77.4%) infants were tested for the influenza virus, of whom 123 (14.3%) tested positive. SBI status was determined in 809 (95.9%) of the 844 infants. Overall, 95 (11.7%) of the 809 infants tested for influenza virus had an SBI. Infants with influenza infections had a significantly lower prevalence of SBI (2.5%) and UTI (2.4%) when compared with infants who tested negative for the influenza virus. Although there were no cases of bacteremia, meningitis, or enteritis in the influenza-positive group, the differences between the 2 groups for these individual infections were not statistically significant. CONCLUSIONS: Febrile infants (1/2)'' and would not receive epinephrine intramuscularly from current auto-injectors. There were 98 children weighing >or=30 kilograms who would receive the 0.3 mg epinephrine auto-injector. Of these 98 children, a total of 29 (30%) had a skin to muscle surface distance of >(5/8)'' and would not receive epinephrine intramuscularly. CONCLUSION: The needle on epinephrine auto injectors is not long enough to reach the muscle in a significant number of children. Increasing the needle length on the auto-injectors would increase the likelihood that more children receive epinephrine by the recommended intramuscular route. PMID- 19564285 TI - Do stimulants protect against psychiatric disorders in youth with ADHD? A 10-year follow-up study. AB - OBJECTIVE: Little is known about the effect of stimulant treatment in youth with attention-deficit/hyperactivity disorder (ADHD) on the subsequent development of comorbid psychiatric disorders. We tested the association between stimulant treatment and the subsequent development of psychiatric comorbidity in a longitudinal sample of patients with ADHD. METHODS: We conducted a case-control, 10-year prospective follow-up study into young-adult years of youth with ADHD. At baseline, we assessed consecutively referred white male children with (n = 140) and without (n = 120) ADHD, aged 6 to 18 years. At the 10-year follow-up, 112 (80%) and 105 (88%) of the children in the ADHD and control groups, respectively, were reassessed (mean age: 22 years). We examined the association between stimulant treatment in childhood and adolescence and subsequent comorbid disorders and grade retention by using proportional hazards survival models. RESULTS: Of the 112 participants with ADHD, 82 (73%) were previously treated with stimulants. Participants with ADHD who were treated with stimulants were significantly less likely to subsequently develop depressive and anxiety disorders and disruptive behavior and less likely to repeat a grade compared with participants with ADHD who were not treated. CONCLUSIONS: We found evidence that stimulant treatment decreases the risk for subsequent comorbid psychiatric disorders and academic failure in youth with ADHD. PMID- 19564286 TI - Epidemiology of sexually transmitted infections in suspected child victims of sexual assault. AB - OBJECTIVE: The objective of this study was to describe the epidemiology of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Treponema pallidum, HIV, and herpes simplex virus type 2 (HSV-2) infection diagnosed by culture or by serologic or microscopic tests and by nucleic acid amplification tests in children who are evaluated for sexual victimization. METHODS: Children aged 0 to 13 years, evaluated for sexual victimization, who required sexually transmissible infection (STI) testing were enrolled at 4 US tertiary referral centers. Specimens for N gonorrhoeae and C trachomatis cultures, wet mounts for detection of T vaginalis, and serologic tests for syphilis and HIV were collected and processed according to study sites' protocols. Nucleic acid amplification tests for C trachomatis and N gonorrhoeae and serologic tests for HSV-2 were performed blinded to other data. RESULTS: Of 536 children enrolled, 485 were female. C trachomatis was detected in 15 (3.1%) and N gonorrhoeae in 16 (3.3%) girls. T vaginalis was identified in 5 (5.9%) of 85 girls by wet mount, 1 (0.3%) of 384 children had a positive serologic screen for syphilis, and 0 of 384 had serologic evidence of HIV infection. Of 12 girls who had a specimen for HSV-2 culture, 5 (41.7%) had a positive result; 7 (2.5%) of 283 had antibody evidence of HSV-2 infection. Overall, 40 (8.2%) of 485 girls and 0 of 51 boys (P = .02) had >or=1 STI. Girls with vaginal discharge were more likely to test positive for an STI (13 [24.5%] of 53) than other girls (27 [6.3%] of 432; prevalence ratio = 3.9; P < .001), although 10 girls with STIs had normal physical examinations. Most girls (27 [67.5%]) with a confirmed STI had normal or nonspecific findings on anogenital examination. CONCLUSIONS: The prevalence of each STI among sexually victimized children is <10%, even when highly sensitive detection methods are used. Most children with STIs have normal or nonspecific findings on physical examination. PMID- 19564287 TI - Incidence and outcomes of pediatric acute lung injury. AB - OBJECTIVE: This population-based, prospective, cohort study was designed to determine the population incidence and outcomes of pediatric acute lung injury. METHODS: Between 1999 and 2000, 1 year of screening was performed at all hospitals admitting critically ill children in King County, Washington. County residents 0.5 to 15 years of age who required invasive (through endotracheal tube or tracheostomy) or noninvasive (through full face mask) mechanical ventilation, regardless of the duration of mechanical ventilation, were screened. From this population, children meeting North American-European Consensus Conference acute lung injury criteria were eligible for enrollment. Postoperative patients who received mechanical ventilation for <24 hours were excluded. Data collected included the presence of predefined cardiac conditions, demographic and physiological data, duration of mechanical ventilation, and deaths. US Census population figures were used to estimate incidence. Associations between outcomes and subgroups identified a priori were assessed. RESULTS: Thirty-nine children met the criteria for acute lung injury, resulting in a calculated incidence of 12.8 cases per 100000 person-years. Severe sepsis (with pneumonia as the infection focus) was the most common risk factor. The median 24-hour Pediatric Risk of Mortality III score was 9.0, and the mean +/- SD was 11.7 +/- 7.5. The hospital mortality rate was 18%, lower than that reported previously for pediatric acute lung injury. There were no statistically significant associations between age, gender, or risk factors and outcomes. CONCLUSIONS: We present the first population-based estimate of pediatric acute lung injury incidence in the United States. Population incidence and mortality rates are lower than those for adult acute lung injury. Low mortality rates in pediatric acute lung injury may necessitate clinical trial outcome measures other than death. PMID- 19564288 TI - Cancer risk among children with very low birth weights. AB - OBJECTIVE: The risk of hepatoblastoma is strongly increased among children with very low birth weight (<1500 g). Because data on very low birth weight and other childhood cancers are sparse, we examined the risk of malignancy with very low birth weight in a large data set. METHODS: We combined case-control data sets created by linking the cancer and birth registries of California, Minnesota, New York, Texas, and Washington states, which included 17672 children diagnosed as having cancer at 0 to 14 years of age and 57966 randomly selected control subjects. Unconditional logistic regression analysis was used to examine the association of cancer with very low birth weight and moderately low birth weight (1500-1999 g and 2000-2499 g, respectively), compared with moderate/high birth weight (>or=2500 g), with adjustment for gender, gestational age, birth order, plurality, maternal age, maternal race, state, and year of birth. RESULTS: Most childhood cancers were not associated with low birth weights. However, retinoblastomas and gliomas other than astrocytomas and ependymomas were possibly associated with very low birth weight. The risk of other gliomas was also increased among children weighing 1500 to 1999 g at birth. CONCLUSIONS: These data suggested no association between most cancers and very low birth weight, with the exception of the known association of hepatoblastoma and possibly moderately increased risks of other gliomas and retinoblastoma, which may warrant confirmation. PMID- 19564289 TI - Convalescent care of infants in the neonatal intensive care unit in community hospitals: risk or benefit? AB - OBJECTIVE: To compare very low birth weight (VLBW) infants transported to a community hospital (CH) before discharge with infants who received convalescent care in a regional-referral NICU (RR-NICU) on 4 parameters: health indicators at the time of hospital discharge, health care use during the 4 months after discharge to home, parent satisfaction with hospital care, and cost of hospitalization. PATIENTS AND METHODS: VLBW infants cared for in 2 RR-NICUs during 2004-2006 were enrolled in the study. One RR-NICU transfers infants to a CH for convalescent care and the other discharges infants directly home. Infants were followed prospectively. Information was gathered from medical charts, parent interviews, and hospital business offices. RESULTS: A total of 255 VLBW infants were enrolled in the study, and 148 were transferred to 15 CHs. Nineteen percent of transferred infants were readmitted to a higher level of care before discharge from the hospital. Preventative health measures and screening examinations were more frequently missed, readmission within 2 weeks of discharge from the hospital was more frequent, parents were less satisfied with hospital care, and duration of hospitalization was 12 days longer, although not statistically different, if infants were transferred to a CH for convalescence rather than discharged from the RR-NICU. Total hospital charges did not differ significantly between the groups. CONCLUSION: Transfer of infants to a CH from an RR-NICU for convalescent care has become routine but may place infants at risk. Our study indicates room for improvement by both CHs and RR-NICUs in the care of transferred VLBW infants. PMID- 19564291 TI - Are outcomes and care processes for preterm neonates influenced by health insurance status? AB - OBJECTIVE: The purpose of this work was to compare the processes of care and to evaluate outcomes of premature neonates delivered to women with Medicaid managed care versus private insurance. DESIGN/METHODS: All of the infants born at <37 weeks' gestation between January 2001 and August 2005 in the ParadigmHealth database were included in these analyses (n = 24151). Infants were categorized by maternal health insurance status as private insurance or Medicaid managed care and analyzed for differences in demographic data and length of stay. For survivors, differences in respiratory care, nutritional, and maturational milestones were assessed. In addition, age to wean to open crib, weight gain, home oxygen, and apnea monitor use were compared. Adverse outcomes, including necrotizing enterocolitis, sepsis, severe intraventricular hemorrhage, severe retinopathy of prematurity, bronchopulmonary dysplasia, apnea, and mortality, were compared. Statistical tests used were Students t test, chi(2), and Kruskall Wallis test. Multiple logistic regression was performed after controlling for demographic variables. RESULTS: Of the 24151 infants studied, 19046 (78.9%) had private insurance, and 5105 (21.1%) had Medicaid managed care. There were no differences in gestational age at birth; however, Medicaid managed care infants had lower birth weight, lower Apgar score at 5 minutes, increased incidence of necrotizing enterocolitis and bacterial sepsis, and longer length of stay. Of the surviving infants, more neonates with private insurance went home on oxygen and apnea monitors despite no differences found in the incidences of apnea or bronchopulmonary dysplasia between the groups. There were no differences in processes of care for feeding and respiratory milestones, but infants with Medicaid managed care weaned to an open crib later and had greater overall weight gain compared with infants with private insurance. CONCLUSIONS: We speculate that, in addition to the known impact of insurance status on well-being at birth, Medicaid managed care is independently associated with adverse neonatal outcomes in preterm infants, as well as differences in neonatal intensive care discharge processes. PMID- 19564292 TI - Morbidities and hospital resource use during the first 3 years of life among very preterm infants. AB - OBJECTIVE: The objective of this study was to determine how the use of hospital resources during the first 3 years of life was associated with prematurity related morbidity in very preterm infants (gestational age of <32 weeks or birth weight of <1501 g). METHODS: The study was a retrospective, national register study including all very preterm infants born alive in Finland between 2000 and 2003 (N = 2148). Infants who died before the age of 3 years (n = 264) or who had missing register data (n = 88) were excluded from the study. The relationship between 6 morbidity groups and the need for hospital care during the first 3 years of life was studied by using a negative binomial model. RESULTS: A total of 66.2% of the infants did not have any of the morbidities studied. Infants who were subsequently diagnosed as having cerebral palsy (6.1% of the study group), later obstructive airway disease (20.0%), hearing loss (2.5%), visual disturbances or blindness (3.8%), or other ophthalmologic problems (13.4%) had initial hospital stays that were a mean of 7, 8, 12, 17, and 3 days longer, respectively, than those for infants without these conditions. All morbidity groups were associated with increased numbers of hospital visits during either the second or third year of life, compared with infants without these morbidities. The need for hospitalizations and outpatient hospital care decreased with postnatal age for infants with later morbidities and for infants without later morbidities. CONCLUSIONS: Most very preterm infants born in Finland survived without severe morbidities and required relatively little hospital care after the initial discharge. However, those with later morbidities had a long initial length of stay and more readmissions and outpatient visits during the 3 year follow-up period. PMID- 19564290 TI - Unimpaired outcomes for extremely low birth weight infants at 18 to 22 months. AB - OBJECTIVE: The goal was to identify, among extremely low birth weight (or=85, normal neurologic examination findings, and normal vision, hearing, swallowing, and walking. Outcomes were determined for 5250 (86%) of 6090 extremely low birth weight inborn infants. RESULTS: Of the 5250 infants whose outcomes were known at 18 months, 850 (16%) were unimpaired, 1153 (22%) had mild impairments, 1147 (22%) had moderate/severe neurodevelopmental impairments, and 2100 (40%) had died. Unimpaired survival rates varied according to birth weight, from <1% for infants or=85th percentile and surveyed their pediatric clinicians (N = 75). The main outcome was parental confidence in their ability to make overweight-related behavior changes. We derived a continuous parental confidence score from 6 questions (Cronbach's alpha = 0.72) regarding parental confidence in limiting television viewing, removing televisions from children's bedrooms, reducing fast-food intake, reducing sugar-sweetened beverage intake, increasing physical activity, and improving overall eating patterns for their family. We used multiple linear regression to predict the effects of parent, child, and clinician characteristics on the parents' confidence scores. RESULTS: The mean (SD) score on the parental confidence scale was 13.0 (3.5), and the range was 0.0 to 24.0. In multivariable analyses, parents who said their clinicians assessed their confidence (41%; beta = 0.73 [95% confidence interval: 0.04-1.42]) or who said that their clinicians assessed their readiness to change (35%; beta =0.80 [95% confidence interval: 0.10-1.49]) reported higher levels of confidence compared with parents whose clinicians did not assess confidence or readiness to change. CONCLUSIONS: Clinician assessment of parental confidence and readiness to change was associated with higher parent confidence in making changes to keep their child from being overweight. PMID- 19564296 TI - Impact of electronic health record-based alerts on influenza vaccination for children with asthma. AB - OBJECTIVE: The goal was to assess the impact of influenza vaccine clinical alerts on missed opportunities for vaccination and on overall influenza immunization rates for children and adolescents with asthma. METHODS: A prospective, cluster randomized trial of 20 primary care sites was conducted between October 1, 2006, and March 31, 2007. At intervention sites, electronic health record-based clinical alerts for influenza vaccine appeared at all office visits for children between 5 and 19 years of age with asthma who were due for vaccine. The proportion of captured immunization opportunities at visits and overall rates of complete vaccination for patients at intervention and control sites were compared with those for the previous year, after standardization for relevant covariates. The study had >80% power to detect an 8% difference in the change in rates between the study and baseline years at intervention versus control practices. RESULTS: A total of 23 418 visits and 11 919 children were included in the study year and 21 422 visits and 10 667 children in the previous year. The majority of children were male, 5 to 9 years of age, and privately insured. With standardization for selected covariates, captured vaccination opportunities increased from 14.4% to 18.6% at intervention sites and from 12.7% to 16.3% at control sites, a 0.3% greater improvement. Standardized influenza vaccination rates improved 3.4% more at intervention sites than at control sites. The 4 practices with the greatest increases in rates (>or=11%) were all in the intervention group. Vaccine receipt was more common among children who had been vaccinated previously, with increasing numbers of visits, with care early in the season, and at preventive versus acute care visits. CONCLUSIONS: Clinical alerts were associated with only modest improvements in influenza vaccination rates. PMID- 19564297 TI - Effects of oseltamivir on influenza-related complications in children with chronic medical conditions. AB - OBJECTIVE: This study investigated the influence of oseltamivir on influenza related complications and hospitalizations for children and adolescents, 1 to 17 years of age, with chronic medical conditions or neurologic or neuromuscular disease. METHODS: In a retrospective study, outcomes for patients who were given oseltamivir within 1 day after influenza diagnosis were compared with those for patients who received no antiviral therapy. Anonymous data from MarketScan databases (Thomson Reuters, Cambridge, MA) were used to identify patients from 6 influenza seasons between 2000 and 2006. The study outcomes were frequencies of pneumonia, respiratory illnesses other than pneumonia, otitis media, and hospitalization. RESULTS: Oseltamivir was prescribed for 1634 patients according to the study criteria, and 3721 patients received no antiviral therapy for their influenza. After adjustment for demographic and medical history variables, oseltamivir was associated with significant reductions in the risks of respiratory illnesses other than pneumonia, otitis media and its complications, and all-cause hospitalization in the 14 days after influenza diagnosis. Analyses for 30 days after influenza diagnosis also showed significant risk reductions for respiratory illnesses other than pneumonia, otitis media and its complications, and all-cause hospitalization with oseltamivir. CONCLUSION: When it was prescribed at influenza diagnosis, oseltamivir was associated with reduced risks of influenza-related complications and hospitalizations for children and adolescents at high risk of influenza complications. PMID- 19564298 TI - Effect of a high-flow open nasal cannula system on obstructive sleep apnea in children. AB - OBJECTIVE: Obstructive sleep apnea syndrome in children is associated with significant morbidity. Continuous positive airway pressure (CPAP) treats obstructive apnea in children, but is impeded by low adherence. We, therefore, sought to assess the effect of warm humidified air delivered through an open nasal cannula (treatment with nasal insufflation [TNI]) on obstructive sleep apnea in children with and without adenotonsillectomy. METHODS: Twelve participants (age: 10 +/- 1 years; BMI: 35 +/- 14 kg/m(2)), with obstructive apnea-hypopnea syndrome ranging from mild to severe (2-36 events per hour) were administered 20 L/min of air through a nasal cannula. Standard sleep architecture, sleep-disordered breathing, and arousal indexes were assessed at baseline, on TNI, and on CPAP. Additional measures of the percentage of time with inspiratory flow limitation, respiratory rate, and inspiratory duty cycle were assessed at baseline and on TNI. RESULTS: TNI reduced the amount of inspiratory flow limitation, which led to a decrease in respiratory rate and inspiratory duty cycle. TNI improved oxygen stores and decreased arousals, which decreased the occurrence of obstructive apnea from 11 +/- 3 to 5 +/- 2 events per hour (P < .01). In the majority of children, the reduction in the apnea-hypopnea index on TNI was comparable to that on CPAP. CONCLUSIONS: TNI offers an alternative to therapy to CPAP in children with mild-to-severe sleep apnea. Additional studies will be needed to determine the efficacy of this novel form of therapy. PMID- 19564299 TI - Chronic fatigue syndrome after infectious mononucleosis in adolescents. AB - OBJECTIVE: The goal was to characterize prospectively the course and outcome of chronic fatigue syndrome in adolescents during a 2-year period after infectious mononucleosis. METHODS: A total of 301 adolescents (12-18 years of age) with infectious mononucleosis were identified and screened for nonrecovery 6 months after infectious mononucleosis by using a telephone screening interview. Nonrecovered adolescents underwent a medical evaluation, with follow-up screening 12 and 24 months after infectious mononucleosis. After blind review, final diagnoses of chronic fatigue syndrome at 6, 12, and 24 months were made by using established pediatric criteria. RESULTS: Six, 12, and 24 months after infectious mononucleosis, 13%, 7%, and 4% of adolescents, respectively, met the criteria for chronic fatigue syndrome. Most individuals recovered with time; only 2 adolescents with chronic fatigue syndrome at 24 months seemed to have recovered or had an explanation for chronic fatigue at 12 months but then were reclassified as having chronic fatigue syndrome at 24 months. All 13 adolescents with chronic fatigue syndrome 24 months after infectious mononucleosis were female and, on average, they reported greater fatigue severity at 12 months. Reported use of steroid therapy during the acute phase of infectious mononucleosis did not increase the risk of developing chronic fatigue syndrome. CONCLUSIONS: Infectious mononucleosis may be a risk factor for chronic fatigue syndrome in adolescents. Female gender and greater fatigue severity, but not reported steroid use during the acute illness, were associated with the development of chronic fatigue syndrome in adolescents. Additional research is needed to determine other predictors of persistent fatigue after infectious mononucleosis. PMID- 19564301 TI - Early nasal continuous positive airway pressure and necrotizing enterocolitis in preterm infants. AB - BACKGROUND: The use of early nasal continuous positive airway pressure (ENCPAP) as the mode of initial respiratory support for very low birth weight (VLBW) infants has been increasing. The impact of CPAP and oxygen on gut mucosa and perfusion in premature infants is not known. The relation between ENCPAP and necrotizing enterocolitis (NEC) has not been adequately addressed. OBJECTIVE: To evaluate if the use of an individualized respiratory management strategy encouraging the use of ENCPAP is associated with an increased risk of NEC, and to determine risk factors for NEC in premature infants supported by CPAP. METHODS: A retrospective analysis was conducted on VLBW infants (birth weight < 1500 g) managed at 2 institutions that use an ENCPAP respiratory management strategy for premature infants. Data on the use of oxygen and mode of ventilatory support were collected during the first 3 days of life. Diagnosis of NEC was used as the dependent variable in a logistic regression model. Birth weight, gender, prenatal steroid use, mode of respiratory support (CPAP versus ventilator) and fraction of inspired oxygen, umbilical artery catheter placement, partial pressure of oxygen, patent ductus arteriosus, early sepsis, hospital, and delivery room management (ENCPAP versus initial intubation) were controlled for in the model. RESULTS: Data on 343 premature infants were collected for this study. Mean birth weight was 999 +/- 289 g and gestational age was 28 +/- 2.6 weeks. The majority of patients were managed with ENCPAP, with only 13% of patients intubated in the delivery room. The overall incidence of NEC was 7% (n = 24). The exposure to ENCPAP did not increase the risk for NEC compared with the use of a ventilator. CONCLUSIONS. The risk of NEC in VLBW premature infants was not increased by the use of ENCPAP. Initial respiratory support with ENCPAP seems to be a safe alternative to routine intubation and mechanical ventilation in premature infants. PMID- 19564300 TI - Low-income parents' views on the redesign of well-child care. AB - OBJECTIVE: To examine the perspectives of low-income parents on redesigning well child care (WCC) for children aged 0 to 3 years, focusing on possible changes in 3 major domains: providers, locations, and formats. METHODS: Eight focus groups (4 English and 4 Spanish) were conducted with 56 parents of children aged 6 months to 5 years, recruited through a federally qualified health center. Discussions were recorded, transcribed, and analyzed by using the constant comparative method of qualitative analysis. RESULTS: Parents were mostly mothers (91%), nonwhite (64% Latino, 16% black), and <30 years of age (66%) and had an annual household income of <$35000 (96%). Parents reported substantial problems with WCC, focusing largely on limited provider access (especially with respect to scheduling and transportation) and inadequate behavioral/developmental services. Most parents endorsed nonphysician providers and alternative locations and formats as desirable adjuncts to usual physician-provided, clinic-based WCC. Nonphysician providers were viewed as potentially more expert in behavioral/developmental issues than physicians and more attentive to parent provider relationships. Some alternative locations for care (especially home and day care visits) were viewed as creating essential context for providers and dramatically improving family convenience. Alternative locations whose sole advantage was convenience (eg, retail-based clinics), however, were viewed more skeptically. Among alternative formats, group visits in particular were seen as empowering, turning parents into informal providers through mutual sharing of behavioral/developmental advice and experiences. CONCLUSIONS: Low-income parents of young children identified major inadequacies in their WCC experiences. To address these problems, they endorsed a number of innovative reforms that merit additional investigation for feasibility and effectiveness. PMID- 19564302 TI - Intravenous lipid and bilirubin-albumin binding variables in premature infants. AB - BACKGROUND: The lipid intake at which a significant bilirubin-displacing effect occurs as a function of gestational age (GA) is unclear. OBJECTIVE: To determine the effect of gradual increase in IL intake from 1.5 to 3 g/kg per day on bilirubin-albumin binding variables as a function of GA in premature infants with indirect hyperbilirubinemia. METHODS: Infants of 24 to 33 weeks' gestation at birth who received IL (20% Intralipid [Fresenius Kabi, Uppsala, Sweden]) doses of 1.5, 2, 2.5, and 3 g/kg per day over 4 consecutive days were prospectively evaluated. The blood samples were drawn twice at least 8 hours apart on each IL intake to measure total serum bilirubin and free bilirubin by the peroxidase test. The highest free bilirubin on each IL intake, the corresponding total serum bilirubin, and serum albumin were used to calculate the bilirubin/albumin binding constant or binding affinity. RESULTS: Sixty-two infants (median GA: 28 weeks) were studied during the first 10 days of life. None of the subjects had culture proven sepsis, had triglyceride levels of >2.05 mmol/L, or were receiving steroids. Infants were grouped in 2-week GA intervals. The cumulative frequency of elevated free bilirubin concentration (>or=90th percentile or B(f) >or= 32 nmol/L) as a function of IL intake was inversely related to GA and was significantly different among 2-week GA groups. There was significant decrease in binding affinity and increase in free bilirubin concentration with higher IL intake for 28 week GA groups. CONCLUSIONS: The IL intake may be associated with a significant fall in the binding affinity of bilirubin for plasma protein and a concomitant increase in free bilirubin concentration in premature infants. The lipid intake at which this occurs depends on GA. PMID- 19564303 TI - Sleep-disordered breathing and behaviors of inner-city children with asthma. AB - OBJECTIVE: To explore the relationship between sleep-disordered breathing (SDB) and behavioral problems among inner-city children with asthma. METHODS: We examined data for 194 children (aged 4-10 years) who were enrolled in a school based asthma intervention program (response rate: 72%). SDB was assessed by using the Sleep-Related Breathing Disorder Questionnaire that contains 3 subscales: snoring, sleepiness, and attention/hyperactivity. For the current study, we modified the Sleep-Related Breathing Disorder Questionnaire by removing the 6 attention/hyperactivity items. A sleep score of >0.33 was considered indicative of SDB. To assess behavior, caregivers completed the Behavior Problem Index (BPI), which includes 8 behavioral subdomains. We conducted bivariate analyses and multiple linear regression to determine the association of SDB with BPI scores. RESULTS: The majority of children (mean age: 8.2 years) were male (56%), black (66%), and insured by Medicaid (73%). Overall, 33% of the children experienced SDB. In bivariate analyses, children with SDB had significantly higher (worse) behavior scores compared with children without SDB on total BPI (13.7 vs 8.8) and the subdomains externalizing (9.4 vs 6.3), internalizing (4.4 vs 2.5), anxious/depressed (2.4 vs 1.3), headstrong (3.2 vs 2.1), antisocial (2.3 vs 1.7), hyperactive (3.0 vs 1.8), peer conflict (0.74 vs 0.43), and immature (2.0 vs 1.5). In multiple regression models adjusting for several important covariates, SDB remained significantly associated with total BPI scores and externalizing, internalizing, anxious/depressed, headstrong, and hyperactive behaviors. Results were consistent across SDB subscales (snoring, sleepiness). CONCLUSIONS: We found that poor sleep was independently associated with behavior problems in a large proportion of urban children with asthma. Systematic screening for SDB in this high-risk population might help to identify children who would benefit from additional intervention. PMID- 19564304 TI - Side effects of methylphenidate in childhood cancer survivors: a randomized placebo-controlled trial. AB - OBJECTIVES: To investigate the frequency and severity of side effects of methylphenidate among childhood survivors of acute lymphoblastic leukemia and brain tumors and identify predictors of higher adverse effect levels. METHODS: Childhood cancer survivors (N = 103) identified as having attention and learning problems completed a randomized, double-blind, 3-week, home-crossover trial of placebo, low-dose methylphenidate (0.3 mg/kg; 10 mg twice daily maximum) and moderate-dose methylphenidate (0.6 mg/kg; 20 mg twice daily maximum). Caregivers completed the Barkley Side Effects Rating Scale (SERS) at baseline and each week during the medication trial. Siblings of cancer survivors (N = 49) were recruited as a healthy comparison group. RESULTS: There was a significantly higher number and severity of symptoms endorsed on the SERS when patients were taking moderate dose compared with placebo or low dose, but not low dose compared with placebo. The number of side effects endorsed on the SERS was significantly lower during all 3 home-crossover weeks (placebo, low dose, moderate dose) when compared with baseline symptom scores. The severity of side effects was also significantly lower, compared with baseline screening, during placebo and low-dose weeks but not moderate-dose weeks. Both the number and severity of symptoms endorsed at baseline were significantly higher for patients compared with siblings. Female gender and lower IQ were associated with higher adverse effect levels. CONCLUSIONS: Methylphenidate is generally well tolerated by childhood cancer survivors. There is a subgroup at increased risk for side effects that may need to be closely monitored or prescribed a lower medication dose. The seemingly paradoxical findings of increased "side effects" at baseline must be considered when monitoring side effects and designing clinical trials. PMID- 19564305 TI - Delivery indications at late-preterm gestations and infant mortality rates in the United States. AB - OBJECTIVE: The rate of preterm births has been increasing in the United States, especially for births 34 to 36 weeks of gestation (late preterm), which now constitute 71% of all preterm births. The causes for these trends remain unclear. We characterized the delivery indications for late preterm births and their potential impact on neonatal and infant mortality rates. PATIENTS AND METHODS: Using the 2001 US Birth Cohort Linked birth/death files of 3 483 496 singleton births, we categorized delivery indications as follows: (1) maternal medical conditions; (2) obstetric complications; (3) major congenital anomalies; (4) isolated spontaneous labor: vaginal delivery without induction and without associated medical/obstetric factors; and (5) no recorded indication. RESULTS: Of the 292 627 late-preterm births, the first 4 categories (those with indications and isolated spontaneous labor) accounted for 76.8%. The remaining 23.2% (67 909) were classified as deliveries with no recorded indication. Factors significantly increasing the chance of no recorded indication were older maternal age; non Hispanic, white mother; >/=13 years of education; Southern, Midwestern, and Western region; multiparity; or previous infant with a >/=4000-g birth weight. The neonatal and infant mortality rates were significantly higher among deliveries with no recorded indication compared with deliveries secondary to isolated spontaneous labor but lower compared with deliveries with an obstetric indication or congenital anomaly. CONCLUSIONS: A total of 23% of late preterm births had no recorded indication for delivery noted on birth certificates. Patient factors may be playing a role in these deliveries. It is concerning that these infants had higher mortality rates compared with those born after spontaneous labor at similar gestational ages. Given the excess risk of mortality, patients and providers need to discuss the risks of delivering a preterm infant in the absence of medical indications at 34 to 36 weeks. PMID- 19564307 TI - Functioning at school age of moderately preterm children born at 32 to 36 weeks' gestational age. AB - OBJECTIVE: To study outcome of low-risk moderately preterm birth between 32 and 36/7 weeks' gestation. METHODS: 377 Moderately preterm children (M: 34.7, SD: 1.2 complete weeks), without need for neonatal intensive care and without dysmaturity or congenital malformations, were compared with 182 term children and assessed at eight years (M: 8.9, SD: 0.54). School situation, IQ, sustained attention, behavior problems, and attention-deficit/hyperactivity characteristics were studied. RESULTS: Special education was attended by 7.7% of the moderately preterm children, more than twice the rate of 2.8% in the general Dutch population of this age. Additional exploration for two preterm subgroups of 32 to 33 versus 34 to 36 weeks' gestation showed a need for special education in 9.7% versus 7.3% and a significant difference in grade retention for 30% versus 17%, respectively. Of the children attending mainstream primary schools, grade retention was found in 19% of the preterm versus 8% of the comparison children. Adjusting for maternal education, a group difference of 3 points was found in IQ. The preterm children needed more time for the sustained attention task. The preterm children had more behavior problems (specifically internalizing problems with 27% scoring above the borderline cut-off), as well as more attention deficit/hyperactivity disorder characteristics (specifically attention deficits). CONCLUSIONS: Cognitive and emotional regulation difficulties affect functioning of moderately preterm children, as school problems, a slightly lower IQ, attention and behavioral problems are found when they are compared with term-born children. Identification and monitoring of precursors of these problems at younger age is needed in view of prevention purposes. PMID- 19564306 TI - Apolipoprotein E genotype modifies the risk of behavior problems after infant cardiac surgery. AB - OBJECTIVE: The goal was to evaluate polymorphisms of the APOE gene as modifiers of neurobehavioral outcomes for preschool-aged children with congenital heart defects, after cardiac surgery. METHODS: A prospective observational study with neurodevelopmental evaluation between the fourth and fifth birthdays was performed. Attention and behavioral skills were assessed through parental report. RESULTS: Parents of 380 children completed the neurobehavioral measures. Child Behavior Checklist scores for the pervasive developmental problem scale were in the at-risk or clinically significant range for 15% of the cohort, compared with 9% for the normative data (P < .00001). Attention problem scores were in the at risk or clinically significant range for 12% of the cohort, compared with 7% for the normative data (P = .0002). The Attention-Deficit/Hyperactivity Disorder Rating Scale-IV, Preschool Version, was completed for 378 children; 30% scored in the clinically significant range for inattention and 22% for impulsivity. After adjustment for covariates, the APOE epsilon2 allele was significantly associated with higher scores (worse problems) for multiple Child Behavior Checklist indices, including somatic complaints (P = .009), pervasive developmental problems (P = .032), and internalizing problems (P = .009). In each case, the epsilon4 allele was associated with a better outcome. APOE epsilon2 carriers had impaired social skills, compared with epsilon4 carriers (P = .009). CONCLUSIONS: For preschool-aged children with congenital heart defects requiring surgery, parental rating scales showed an increased prevalence of restricted behavior patterns, inattention, and impaired social interactions. The APOE epsilon2 allele was associated with increased behavior problems, impaired social interactions, and restricted behavior patterns. PMID- 19564309 TI - The association of lung disease with cerebral white matter abnormalities in preterm infants. AB - OBJECTIVE: Preterm infants have a high incidence of neurodevelopmental impairment associated with diffuse cerebral white matter abnormalities and also a high incidence of serious respiratory disease. However, it is unclear if lung disease and brain injury are related, and previous research has been impeded by confounding effects, including prematurity and infection. Using a new approach that permits multivariate statistical analysis, we tested the hypothesis that lung disease is associated with specific white matter abnormalities, detected as reduced fractional anisotropy (FA) in diffusion tensor imaging data. METHODS: Fifty-three preterm infants with no evidence of focal abnormality on conventional MRI were studied at term-equivalent age by using tract-based spatial statistics, an automated observer-independent method for voxelwise analysis of major white matter pathways. RESULTS: In several white matter tracts, FA decreased with a linear relation to the gestational age at birth. Independent of the confounding effects of prematurity and age at scan, respiratory disease was associated with specific white matter abnormalities in preterm infants; those infants receiving mechanical ventilation for >2 days in the perinatal period (n = 10) showed reduced FA in the genu of the corpus callosum, whereas subjects with chronic lung disease (n = 15) displayed a reduction in FA in the left inferior longitudinal fasciculus. CONCLUSION: Independent of the degree of prematurity, respiratory disease is associated with cerebral white matter abnormalities. PMID- 19564310 TI - Carbon monoxide is a significant mediator of cardiovascular status following preterm birth. AB - OBJECTIVE: With male gender as a strong predictor of cardiovascular instability, we hypothesized that gender-specific differences in circulating carbon monoxide levels contributed to dysregulated microvascular function in preterm male infants. METHODS: Infants born at 24 to 34 weeks of gestation (N = 84) were studied in a regional tertiary neonatal unit. Carboxyhemoglobin levels were measured through spectrophotometry in umbilical arterial blood and at 24, 72, and 120 hours after birth. Microvascular blood flow was determined through laser Doppler flowmetry. RESULTS: Carboxyhemoglobin levels demonstrated a strong inverse relationship with gestational age (r = -0.636; P < .001) and were higher in boys (P = .032). Repeated-measures analysis of variance showed a significant decrease in arterial carboxyhemoglobin levels over time (P < .001), with significant between-subjects effects for gestational age (P = .011) and gender (P = .025). Positive correlations with microvascular blood flow at 24 hours of age (r = 0.495; P < .001) and 120 hours of age (r = 0.548; P < .001) were observed. With controlling for gestational age, carboxyhemoglobin levels at 72 hours were greater for infants who died in the first week of life (P = .035). CONCLUSIONS: The gestational age- and gender-specific differences in carboxyhemoglobin levels and the relationship with dysregulated microvascular blood flow, a state related to greater illness severity and hypotension, are novel findings not confined solely to sick preterm infants. Both inducible heme oxygenase-dependent and non heme oxygenase-dependent pathways may initially play a central role in carbon monoxide production, inducing pathophysiologic processes in a gender-specific manner. PMID- 19564308 TI - Risk factors affecting school readiness in premature infants with respiratory distress syndrome. AB - OBJECTIVE: With advances in neonatal care, more children born prematurely are successfully reaching school age. It is unknown how many will be ready for school and what factors affect school readiness. Our objective was to assess readiness of children born prematurely for entry into public school, and determine risk factors associated with lack of school readiness in this population. METHODS: This was a single-center prospective cohort study. Follow- up data were collected for 135 of 167 (81%) surviving premature infants with RDS requiring surfactant replacement therapy. The children were seen between July 2005 and September 2006 (average age: 5.7 +/- 1.0 years) and underwent standardized neurodevelopmental and health assessments and socioeconomic status classification. A 4-level school readiness score was constructed by using each child's standardized scores on assessments of basic concepts (Bracken School-Readiness Assessment), perceptual skills (Visual-Motor Integration Test), receptive vocabulary (Peabody Picture Vocabulary Test, Third Edition), daily living functional skills (Pediatric Functional Independence Measure), and presence of sensory impairments or autism. Proportional odds models were used to identify risk factors predicting lower school-readiness levels. RESULTS: Mean birth weight was 1016 +/- 391 g, and mean gestational age was 27.5 +/- 2.6 weeks. Ninety-one (67%) children were school ready. Using multivariate analysis, male gender, chronic lung disease, and severe intraventricular hemorrhage or periventricular leukomalacia were associated with lower school-readiness levels. However, the most powerful factor determining school-readiness level was low socioeconomic status. CONCLUSION: Interventions targeting neonatal morbidities may be much less effective at improving overall performance at school age compared with the effect of the impoverished social environment. PMID- 19564311 TI - Child protection outcomes for infants of substance-using mothers: a matched cohort study. AB - OBJECTIVE: Parental drug use is a critical public health issue; it is estimated to be present in up to 80% of referrals to Australian child protection agencies. However, no data regarding the child protection outcomes of infants of substance using parents exist in Australia, and no comparisons have been made with infants of non-substance-using parents. We assessed differences in substantiated abuse between 2 groups of mothers in Brisbane to quantify this risk. METHODS: Mothers who disclosed opiate, amphetamine, or methadone use between 2000 and 2003 were identified and compared with non-substance-using mothers who were matched for gender and gestational age. All infants were linked to the Department of Child Safety Child Protection Information System database. Child protection outcomes, such as substantiated notifications and entry into foster care, were compared between groups. RESULTS: We studied 119 infants of substance-using mothers and 238 matched infants. Infants of substance-using mothers were more likely to suffer substantiated harm (hazard ratio 13.3 [95% confidence interval 4.6-38.3]) and to enter foster care (hazard ratio 13.3 [95% confidence interval 5.1-34.3]). Infants of mothers using illicit drugs were more likely to suffer substantiated harm and more likely to enter foster care than infants of mothers who were compliant with a methadone program. CONCLUSIONS: Infants of substance-using mothers have much poorer child protection outcomes than infants of non-substance using mothers. This study adds substantial evidence toward a real association between maternal drug use and child abuse. Greater interagency collaboration is urgently required to reduce this risk. PMID- 19564312 TI - Cerebral oxygen saturation and extraction in preterm infants with transient periventricular echodensities. AB - OBJECTIVE: Our aim was to determine regional cerebral tissue oxygen saturation and fractional tissue oxygen extraction in preterm infants with transient periventricular echodensities. We hypothesized that as a result of reduced cerebral perfusion, regional cerebral tissue oxygen saturation will be lower and fractional tissue oxygen extraction will be higher during the first days after birth. PATIENTS AND METHODS: This was a prospective, observational study of 49 preterm infants (gestational age median: 30.1 weeks [26.0-31.8 weeks]; birth weight median: 1220 g [615-2250 g]). We defined transient periventricular echodensities as echodensities that persisted for >7 days. Regional cerebral tissue oxygen saturation was measured on days 1-5, 8, and 15 after birth. Fractional tissue oxygen extraction was calculated as (transcutaneous arterial oxygen saturation--regional cerebral tissue oxygen saturation)/transcutaneous arterial oxygen saturation. RESULTS: Transient periventricular echodensities were found in 25 of 49 infants. During the first week we found no difference between the 2 groups for cerebral tissue oxygen saturation and fractional tissue oxygen extraction values. On day 15 after birth, cerebral tissue oxygen saturation was lower in preterm infants with transient periventricular echodensities (66%) compared with infants without echodensities (76%) (P = .003). Fractional tissue oxygen extraction in infants with transient periventricular echodensities (0.30) was higher than fractional tissue oxygen extraction in infants without transient periventricular echodensities (0.20) (P < .001). The differences could not be explained by confounding variables. CONCLUSIONS: Persistent transient periventricular echodensities may be associated with increased cerebral oxygen demand after the first week after birth, which is contrary to our hypothesis. Cerebral oxygenation may be involved in the recovery of perinatal white matter damage. PMID- 19564314 TI - Serum gentamicin concentrations in encephalopathic infants are not affected by therapeutic hypothermia. AB - OBJECTIVE: Mild hypothermia for 72 hours is neuroprotective in newborns with moderate or severe hypoxic-ischemic encephalopathy. A core temperature of 33.5 degrees C might reduce drug clearance leading to potential toxicity. Gentamicin is nephrotoxic and ototoxic at high serum concentrations. No study has investigated the influence of 72 hours of hypothermia on serum gentamicin concentrations (SGCs) in children of any age. We aimed to compare the SGCs in encephalopathic infants who underwent intensive care with therapeutic hypothermia or normothermia. METHODS: Data were collected retrospectively from 2 NICUs in Bristol, United Kingdom, that offered cooling therapy within clinical trials since 1998. Eligible infants (n = 55) developed grade 2/3 encephalopathy after birth and fulfilled the entry criteria defined in the CoolCap trial. Encephalopathic infants with similar demographic values were either nursed under normothermia or 72 h-hypothermia. Once-daily gentamicin dosage (4-5 mg/kg) was administered, and trough SGC was recorded with corresponding creatinine concentrations. The time and number of omitted drug doses were noted. RESULTS: Mean trough SGC (pre-second dose) and mean plasma creatinine concentrations for both treatment groups were similar (gentamicin: 2.19 +/- 1.7 [hypothermia] and 2.30 +/- 2.0 [normothermia] mg/L; creatinine: 115.6 +/- 42.8 [hypothermia] and 121.0 +/- 45.1 [normothermia] mumol/L). Forty percent of the trough SGCs in both groups were above the recommended trough concentration of 2.0 mg/L. A significant correlation (r(2) = 0.36) was found between high SGCs and impaired renal function assessed by raised plasma creatinine levels regardless of treatment options. CONCLUSIONS: Our data confirm that impaired renal function is strongly associated with high SGCs. Reduced body temperatures do not affect the clearance of gentamicin. PMID- 19564313 TI - Elevated cerebral pressure passivity is associated with prematurity-related intracranial hemorrhage. AB - OBJECTIVES: Cerebral pressure passivity is common in sick premature infants and may predispose to germinal matrix/intraventricular hemorrhage (GM/IVH), a lesion with potentially serious consequences. We studied the association between the magnitude of cerebral pressure passivity and GM/IVH. PATIENTS AND METHODS: We enrolled infants <32 weeks' gestational age with indwelling mean arterial pressure (MAP) monitoring and excluded infants with known congenital syndromes or antenatal brain injury. We recorded continuous MAP and cerebral near-infrared spectroscopy hemoglobin difference (HbD) signals at 2 Hz for up to 12 hours/day and up to 5 days. Coherence and transfer function analysis between MAP and HbD signals was performed in 3 frequency bands (0.05-0.25, 0.25-0.5, and 0.5-1.0 Hz). Using MAP-HbD gain and clinical variables (including chorioamnionitis, Apgar scores, gestational age, birth weight, neonatal sepsis, and Score for Neonatal Acute Physiology II), we built a logistic regression model that best predicts cranial ultrasound abnormalities. RESULTS: In 88 infants (median gestational age: 26 weeks [range 23-30 weeks]), early cranial ultrasound showed GM/IVH in 31 (37%) and parenchymal echodensities in 10 (12%) infants; late cranial ultrasound showed parenchymal abnormalities in 19 (30%) infants. Low-frequency MAP-HbD gain (highest quartile mean) was significantly associated with early GM/IVH but not other ultrasound findings. The most parsimonious model associated with early GM/IVH included only gestational age and MAP-HbD gain. CONCLUSIONS: This novel cerebrovascular monitoring technique allows quantification of cerebral pressure passivity as MAP-HbD gain in premature infants. High MAP-HbD gain is significantly associated with GM/IVH. Precise temporal and causal relationship between MAP-HbD gain and GM/IVH awaits further study. PMID- 19564315 TI - Triplets across the first 5 years: the discordant infant at birth remains at developmental risk. AB - OBJECTIVES: To examine whether the risk posed to infant development by triplet birth persists into childhood and whether growth-discordant triplets are at a particularly high developmental risk. METHODS: Twenty-one sets of triplets were matched with 21 sets of twins and 21 singletons (N = 126) for medical and demographic conditions and were followed from birth to 5 years. At 6, 12, and 24 months, cognitive development was assessed and mother-infant interactions were coded for maternal sensitivity and child social engagement. At 5 years, the children's cognitive development and neuropsychological skills were tested, social engagement was coded from mother-child interactions, and behavior problems were examined. Maternal adjustment was assessed during interviews at 1 and 5 years. RESULTS: Although triplets showed lower cognitive performance at 6, 12, and 24 months as compared with singletons and twins, differences were attenuated by 5 years in both global IQ and executive functions. Similarly, the lower social engagement observed across infancy and the higher internalizing symptoms reported at 2 years for those in the triplet group were no longer found at 5 years. Difficulties in maternal adjustment among mothers of triplets decreased from 1 to 5 years. However, in 65.2% of the initial sample there was a weight discordance of >15% at birth, and the discordant triplets showed poorer cognitive and social development as compared with their siblings across infancy. At 5 years, the discordant children demonstrated lower cognitive and executive functions performance, decreased social engagement, and higher internalizing symptoms as compared with both siblings and peers. CONCLUSIONS: Whereas most triplets catch up after an early developmental delay, the risk for discordant triplets seems to persist into childhood. Such infants, who are at both biological and environmental risk, should receive special and consistent professional care. PMID- 19564316 TI - Cardiovascular medication errors in children. AB - OBJECTIVES: We sought to describe pediatric cardiovascular medication errors and to determine patients and medications with more-frequently reported and/or more harmful errors. METHODS: We analyzed cardiovascular medication error reports from 2003-2004 for patients <18 years of age, from the US Pharmacopeia MEDMARX database. Reports were stratified according to harm score (A, near miss; B-D, error, no harm; E-I, harmful error). Proportions of harmful reports were determined according to drug class and age group. "High-risk" drugs were defined as antiarrhythmics, antihypertensives, digoxin, and calcium channel blockers. RESULTS: A total of 147 facilities submitted 821 reports with community hospitals predominating (70%). Mean patient age was 4 years (median: 0.9 years). The most common error locations were NICUs, general care units, PICUs, pediatric units, and inpatient pharmacies. Drug administration, particularly improper dosing, was implicated most commonly. Severity analysis showed 5% "near misses," 91% errors without harm, and 4% harmful errors, with no reported fatalities. A total of 893 medications were cited in 821 reports. Diuretics were cited most frequently, followed by antihypertensives, angiotensin inhibitors, beta-adrenergic receptor blockers, digoxin, and calcium channel blockers. Calcium channel blockers, phosphodiesterase inhibitors, antiarrhythmics, and digoxin had the largest proportions of harmful events, although the values were not statistically significantly different from those for other drug classes. Infants <1 year of age accounted for 50% of reports. Proportions of harmful events did not differ according to age. CONCLUSIONS: Infants <1 year of age were most frequently reported in cardiovascular medication errors reaching inpatients, in a national, voluntary, error-reporting database. Proportions of harmful errors were not significantly different by age or cardiovascular medication. Most errors were related to medication administration, largely due to improper dosing. PMID- 19564318 TI - Teaching by listening: the importance of adult-child conversations to language development. AB - OBJECTIVE: To test the independent association of adult language input, television viewing, and adult-child conversations on language acquisition among infants and toddlers. METHODS: Two hundred seventy-five families of children aged 2 to 48 months who were representative of the US census were enrolled in a cross sectional study of the home language environment and child language development (phase 1). Of these, a representative sample of 71 families continued for a longitudinal assessment over 18 months (phase 2). In the cross-sectional sample, language development scores were regressed on adult word count, television viewing, and adult-child conversations, controlling for socioeconomic attributes. In the longitudinal sample, phase 2 language development scores were regressed on phase 1 language development, as well as phase 1 adult word count, television viewing, and adult-child conversations, controlling for socioeconomic attributes. RESULTS: In fully adjusted regressions, the effects of adult word count were significant when included alone but were partially mediated by adult-child conversations. Television viewing when included alone was significant and negative but was fully mediated by the inclusion of adult-child conversations. Adult-child conversations were significant when included alone and retained both significance and magnitude when adult word count and television exposure were included. CONCLUSIONS: Television exposure is not independently associated with child language development when adult-child conversations are controlled. Adult child conversations are robustly associated with healthy language development. Parents should be encouraged not merely to provide language input to their children through reading or storytelling, but also to engage their children in two-sided conversations. PMID- 19564317 TI - Trajectories of receptive language development from 3 to 12 years of age for very preterm children. AB - OBJECTIVES: The goal was to examine whether indomethacin use, gender, neonatal, and sociodemographic factors predict patterns of receptive language development from 3 to 12 years of age in preterm children. METHODS: A total of 355 children born in 1989-1992 with birth weights of 600 to 1250 g were evaluated at 3, 4.5, 6, 8, and 12 years with the Peabody Picture Vocabulary Test-Revised. Hierarchical growth modeling was used to explore differences in language trajectories. RESULTS: From 3 to 12 years, preterm children displayed catch-up gains on the Peabody Picture Vocabulary Test-Revised. Preterm children started with an average standardized score of 84.1 at 3 years and gained 1.2 points per year across the age period studied. Growth-curve analyses of Peabody Picture Vocabulary Test Revised raw scores revealed an indomethacin-gender effect on initial scores at 3 years, with preterm boys assigned randomly to receive indomethacin scoring, on average, 4.2 points higher than placebo-treated boys. However, the velocity of receptive vocabulary development from 3 to 12 years did not differ for the treatment groups. Children with severe brain injury demonstrated slower gains in skills over time, compared with those who did not suffer severe brain injury. Significant differences in language trajectories were predicted by maternal education and minority status. CONCLUSION: Although indomethacin yielded an initial benefit for preterm boys, this intervention did not alter the developmental trajectory of receptive language scores. Severe brain injury leads to long-term sequelae in language development, whereas a socioeconomically advantaged environment supports better language development among preterm children. PMID- 19564319 TI - Maternal allopurinol during fetal hypoxia lowers cord blood levels of the brain injury marker S-100B. AB - BACKGROUND: Fetal hypoxia is an important determinant of neonatal encephalopathy caused by birth asphyxia, in which hypoxia-induced free radical formation plays an important role. HYPOTHESIS: Maternal treatment with allopurinol, will cross the placenta during fetal hypoxia (primary outcome) and reduce S-100B and free radical formation (secondary outcome). METHODS: In a randomized, double-blind feasibility study, 53 pregnant women in labor (54 fetuses) with a gestational age of >36 weeks and fetal hypoxia, as indicated by abnormal/nonreassuring fetal heart rate tracing or fetal scalp pH of <7.20, received 500 mg of allopurinol or placebo intravenously. Severity of fetal hypoxia, brain damage and free radical formation were assessed by arterial cord blood lactate, S-100B and non-protein bound-iron concentrations, respectively. At birth, maternal and cord blood concentrations of allopurinol and its active metabolite oxypurinol were determined. RESULTS: Allopurinol and oxypurinol concentrations were within the therapeutic range in the mother (allopurinol > 2 mg/L and/or oxypurinol > 4 mg/L) but not always in arterial cord blood. We therefore created 3 groups: a placebo (n = 27), therapeutic allopurinol (n = 15), and subtherapeutic allopurinol group (n = 12). Cord lactate concentration did not differ, but S-100B was significantly lower in the therapeutic allopurinol group compared with the placebo and subtherapeutic allopurinol groups (P < .01). Fewer therapeutic allopurinol cord samples had measurable non-protein-bound iron concentrations compared with placebo (P < .01). CONCLUSIONS: Maternal allopurinol/oxypurinol crosses the placenta during fetal hypoxia. In fetuses/newborns with therapeutic allopurinol/oxypurinol concentrations in cord blood, lower plasma levels of the brain injury marker protein S-100B were detected. A larger allopurinol trial in compromised fetuses at term seems warranted. The allopurinol dosage must be adjusted to achieve therapeutic fetal allopurinol/oxypurinol concentrations. PMID- 19564320 TI - Improved outcomes associated with medical home implementation in pediatric primary care. AB - OBJECTIVE: The medical home model with its emphasis on planned care, care coordination, family-centered approaches, and quality provides an attractive concept construct for primary care redesign. Studies of medical home components have shown increased quality and reduced costs, but the medical home model as a whole has not been studied systematically. This study tested the hypothesis that increased medical homeness in primary care practice is associated with decreased utilization of health services and increased patient satisfaction. METHODS: Forty three primary care practices were identified through 7 health plans in 5 states. Using the Medical Home Index (MHI), each practice's implementation of medical home concepts "medical homeness" was measured. Health plans provided the previous year's utilization data for children with 6 chronic conditions. The plans identified 42 children in each practice with these chronic conditions and surveyed their families regarding satisfaction with care and burden of illness. RESULTS: Higher MHI scores and higher subdomain scores for organizational capacity, care coordination, and chronic-condition management were associated with significantly fewer hospitalizations. Higher chronic-condition management scores were associated with lower emergency department use. Family survey data yielded no recognizable trends with respect to the medical home measurement. CONCLUSIONS: Developing an evidence base for the value of the primary care medical home has importance for providers, payers, policy makers, and consumers. Reducing hospitalizations through enhanced primary care provides a potential case for new reimbursement strategies supporting medical home services such as care coordination. Larger-scale studies are needed to further develop/examine these relationships. PMID- 19564321 TI - Improving clinical quality indicators through electronic health records: it takes more than just a reminder. PMID- 19564322 TI - Ethnicity matters in the assessment and treatment of children's pain. PMID- 19564323 TI - Speed isn't everything in pediatric medical transport. PMID- 19564324 TI - The association of high-magnitude cerebral passivity and intraventricular hemorrhage in premature infants. PMID- 19564325 TI - Optimizing early development and understanding trajectories of resiliency after extreme prematurity. PMID- 19564326 TI - Pediatrics print edition is redesigned to better meet your needs. PMID- 19564327 TI - Policy Statement--Using personal health records to improve the quality of health care for children. AB - A personal health record (PHR) is a repository of information from multiple contributors (eg, patient, family, guardians, physicians, and other health care professionals) regarding the health of an individual. The development of electronic PHRs presents new opportunities and challenges to the practice of pediatrics. This policy statement provides recommendations for actions that pediatricians can take to support the development and use of PHRs for children. Pediatric health care professionals must become actively involved in developing and adopting PHRs and PHR systems. The American Academy of Pediatrics supports development of: educational programs for families and clinicians on effective and efficient use of PHRs; incentives to facilitate PHR use and maintenance; and child- and adolescent-friendly standards for PHR content, portability, security, and privacy. Properly designed PHR systems for pediatric care can empower patients. PHRs can improve access to health information, improve coordination of preventive health and health maintenance activities, and support emergency and disaster management activities. PHRs provide support for the medical home for all children, including those with special health care needs and those in foster care. PHRs can also provide information to serve as the basis for pediatric quality improvement efforts. For PHRs to be adopted sufficiently to realize these benefits, we must determine how best to support their development and adoption. Privacy and security issues, especially with regard to children and adolescents, must be addressed. PMID- 19564328 TI - Policy statement--The future of pediatrics: mental health competencies for pediatric primary care. AB - Pediatric primary care clinicians have unique opportunities and a growing sense of responsibility to prevent and address mental health and substance abuse problems in the medical home. In this report, the American Academy of Pediatrics proposes competencies requisite for providing mental health and substance abuse services in pediatric primary care settings and recommends steps toward achieving them. Achievement of the competencies proposed in this statement is a goal, not a current expectation. It will require innovations in residency training and continuing medical education, as well as a commitment by the individual clinician to pursue, over time, educational strategies suited to his or her learning style and skill level. System enhancements, such as collaborative relationships with mental health specialists and changes in the financing of mental health care, must precede enhancements in clinical practice. For this reason, the proposed competencies begin with knowledge and skills for systems-based practice. The proposed competencies overlap those of mental health specialists in some areas; for example, they include the knowledge and skills to care for children with attention-deficit/hyperactivity disorder, anxiety, depression, and substance abuse and to recognize psychiatric and social emergencies. In other areas, the competencies reflect the uniqueness of the primary care clinician's role: building resilience in all children; promoting healthy lifestyles; preventing or mitigating mental health and substance abuse problems; identifying risk factors and emerging mental health problems in children and their families; and partnering with families, schools, agencies, and mental health specialists to plan assessment and care. Proposed interpersonal and communication skills reflect the primary care clinician's critical role in overcoming barriers (perceived and/or experienced by children and families) to seeking help for mental health and substance abuse concerns. PMID- 19564329 TI - Clinical report--Antenatal counseling regarding resuscitation at an extremely low gestational age. AB - The anticipated delivery of an extremely low gestational age infant raises difficult questions for all involved, including whether to initiate resuscitation after delivery. Each institution caring for women at risk of delivering extremely preterm infants should provide comprehensive and consistent guidelines for antenatal counseling. Parents should be provided the most accurate prognosis possible on the basis of all the factors known to affect outcome for a particular case. Although it is not feasible to have specific criteria for when the initiation of resuscitation should or should not be offered, the following general guidelines are suggested. If the physicians involved believe there is no chance for survival, resuscitation is not indicated and should not be initiated. When a good outcome is considered very unlikely, the parents should be given the choice of whether resuscitation should be initiated, and clinicians should respect their preference. Finally, if a good outcome is considered reasonably likely, clinicians should initiate resuscitation and, together with the parents, continually reevaluate whether intensive care should be continued. Whenever resuscitation is considered an option, a qualified individual, preferably a neonatologist, should be involved and should be present in the delivery room to manage this complex situation. Comfort care should be provided for all infants for whom resuscitation is not initiated or is not successful. PMID- 19564330 TI - Policy statement--Expert witness participation in civil and criminal proceedings. AB - The interests of the public and both the medical and legal professions are best served when scientifically sound and unbiased expert witness testimony is readily available in civil and criminal proceedings. As members of the medical community, patient advocates, and private citizens, pediatricians have ethical and professional obligations to assist in the administration of justice. The American Academy of Pediatrics believes that the adoption of the recommendations outlined in this statement will improve the quality of medical expert witness testimony in legal proceedings and, thereby, increase the probability of achieving outcomes that are fair, honest, and equitable. Strategies for enforcing guidance and promoting oversight of expert witnesses are proposed. PMID- 19564331 TI - Phosphorylation-independent regulation of metabotropic glutamate receptor 5 desensitization and internalization by G protein-coupled receptor kinase 2 in neurons. AB - The uncoupling of metabotropic glutamate receptors (mGluRs) from heterotrimeric G proteins represents an essential feedback mechanism that protects neurons against receptor overstimulation that may ultimately result in damage. The desensitization of mGluR signaling is mediated by both second messenger-dependent protein kinases and G protein-coupled receptor kinases (GRKs). Unlike mGluR1, the attenuation of mGluR5 signaling in HEK 293 cells is reported to be mediated by a phosphorylation-dependent mechanism. However, the mechanisms regulating mGluR5 signaling and endocytosis in neurons have not been investigated. Here we show that a 2-fold overexpression of GRK2 leads to the attenuation of endogenous mGluR5-mediated inositol phosphate (InsP) formation in striatal neurons and siRNA knockdown of GRK2 expression leads to enhanced mGluR5-mediated InsP formation. Expression of a catalytically inactive GRK2-K220R mutant also effectively attenuates mGluR5 signaling, but the expression of a GRK2-D110A mutant devoid in Galpha(q/11) binding increases mGluR5 signaling in response to agonist stimulation. Taken together, these results indicate that the attenuation of mGluR5 responses in striatal neurons is phosphorylation-independent. In addition, we find that mGluR5 does not internalize in response to agonist treatment in striatal neuron, but is efficiently internalized in cortical neurons that have higher levels of endogenous GRK2 protein expression. When overexpressed in striatal neurons, GRK2 promotes agonist-stimulated mGluR5 internalization. Moreover, GRK2-mediated promotion of mGluR5 endocytosis does not require GRK2 catalytic activity. Thus, we provide evidence that GRK2 mediates phosphorylation independent mGluR5 desensitization and internalization in neurons. PMID- 19564332 TI - Sequential and compartment-specific phosphorylation controls the life cycle of the circadian CLOCK protein. AB - The circadian clock facilitates a temporal coordination of most homeostatic activities and their synchronization with the environmental cycles of day and night. The core oscillating activity of the circadian clock is formed by a heterodimer of the transcription factors CLOCK (CLK) and CYCLE (CYC). Post translational regulation of CLK/CYC has previously been shown to be crucial for clock function and accurate timing of circadian transcription. Here we report that a sequential and compartment-specific phosphorylation of the Drosophila CLK protein assigns specific localization and activity patterns. Total and nuclear amounts of CLK protein were found to oscillate over the course of a day in circadian neurons. Detailed analysis of the cellular distribution and phosphorylation of CLK revealed that newly synthesized CLK is hypophosphorylated in the cytoplasm prior to nuclear import. In the nucleus, CLK is converted into an intermediate phosphorylation state that correlates with trans-activation of circadian transcription. Hyperphosphorylation and degradation are promoted by nuclear export of the CLK protein. Surprisingly, CLK localized to discrete nuclear foci in cell culture as well as in circadian neurons of the larval brain. These subnuclear sites likely contain a storage form of the transcription factor, while homogeneously distributed nuclear CLK appears to be the transcriptionally active form. These results show that sequential post-translational modifications and subcellular distribution regulate the activity of the CLK protein, indicating a core post-translational timing mechanism of the circadian clock. PMID- 19564333 TI - Detergent-activated BAX protein is a monomer. AB - BAX is a pro-apoptotic member of the BCL-2 protein family. At the onset of apoptosis, monomeric, cytoplasmic BAX is activated and translocates to the outer mitochondrial membrane, where it forms an oligomeric pore. The chemical mechanism of BAX activation is controversial, and several in vitro and in vivo methods of its activation are known. One of the most commonly used in vitro methods is activation with detergents, such as n-octyl glucoside. During BAX activation with n-octyl glucoside, it has been shown that BAX forms high molecular weight complexes that are larger than the combined molecular weight of BAX monomer and one detergent micelle. These large complexes have been ascribed to the oligomerization of BAX prior to its membrane insertion and pore formation. This is in contrast to the in vivo studies that suggest that active BAX inserts into the outer mitochondrial membrane as a monomer and then undergoes oligomerization. Here, to simultaneously determine the molecular weight and the number of BAX proteins per BAX-detergent micelle during detergent activation, we have used an approach that combines two single-molecule sensitivity technique, fluorescence correlation spectroscopy, and fluorescence-intensity distribution analysis. We have tested a range of detergents as follows: n-octyl glucoside, dodecyl maltoside, Triton X-100, Tween 20, 3-[(3-cholamidopropyl)dimethylammonio]-1 propanesulfonic acid, and cholic acid. With these detergents we observe that BAX is a monomer before, during, and after interaction with micelles. We conclude that detergent activation of BAX is not congruent with oligomerization and that in physiologic buffer conditions BAX can assume two stable monomeric conformations, one inactive and one active. PMID- 19564334 TI - Cyclin-dependent kinase 2-associating protein 1 commits murine embryonic stem cell differentiation through retinoblastoma protein regulation. AB - Mouse embryonic stem cells (mESCs) maintain pluripotency and indefinite self renewal through yet to be defined molecular mechanisms. Leukemia inhibitory factor has been utilized to maintain the symmetrical self-renewal and pluripotency of mESCs in culture. It has been suggested that molecules with significant cellular effects on retinoblastoma protein (pRb) or its related pathways should have functional impact on mESC proliferation and differentiation. However, the involvement of pRb in pluripotent differentiation of mESCs has not been extensively elaborated. In this paper, we present novel experimental data indicating that Cdk2ap1 (cyclin-dependent kinase 2-associating protein 1), an inhibitor of G(1)/S transition through down-regulation of CDK2 and an essential gene for early embryonic development, confers competency for mESC differentiation. Targeted disruption of Cdk2ap1 in mESCs resulted in abrogation of leukemia inhibitory factor withdrawal-induced differentiation, along with altered pRb phosphorylation. The differentiation competency of the Cdk2ap1(-/-) mESCs was restored upon the ectopic expression of Cdk2ap1 or a nonphosphorylatable pRb mutant (mouse Ser(788) --> Ala), suggesting that the CDK2AP1-mediated differentiation of mESCs was elicited through the regulation of pRb. Further analysis on mESC maintenance or differentiation-related gene expression supports the phosphorylation at serine 788 in pRb plays a significant role for the CDK2AP1-mediated differentiation of mESCs. These data clearly demonstrate that CDK2AP1 is a competency factor in the proper differentiation of mESCs by modulating the phosphorylation level of pRb. This sheds light on the role of the establishment of the proper somatic cell type cell cycle regulation for mESCs to enter into the differentiation process. PMID- 19564335 TI - Hypoxia-induced expression of carbonic anhydrase 9 is dependent on the unfolded protein response. AB - Adaptation to tumor hypoxia is mediated in large part by changes in protein expression. These are driven by multiple pathways, including activation of the hypoxia inducible factor-1 (HIF-1) transcription factor and the PKR-like endoplasmic reticulum kinase PERK, a component of the unfolded protein response. Through gene expression profiling we discovered that induction of the HIF-1 target gene CA9 was defective in mouse embryo fibroblasts derived from mice harboring an eIF2alpha S51A knock-in mutation. This finding was confirmed in two isogenic human cell lines with an engineered defect in eIF2alpha phosphorylation. We show that impaired CA9 expression was not due to changes in HIF activity or CA9 mRNA stability. Using chromatin immunoprecipitation we show that the eIF2alpha-dependent translationally regulated gene ATF4 binds directly to the CA9 promoter and is associated with loss of the transcriptional repressive histone 3 lysine 27 tri-methylation mark. Loss or overexpression of ATF4 confirmed its role in CA9 induction during hypoxia. Our data indicate that expression of CA9 is regulated through both the HIF-1 and unfolded protein response hypoxia response pathways in vitro and in vivo. PMID- 19564336 TI - RGS5, a hypoxia-inducible apoptotic stimulator in endothelial cells. AB - Endothelial cells rapidly respond to changes in oxygen homeostasis by regulating gene expression. Regulator of G protein signaling 5 (RGS5) is a negative regulator of G protein-mediated signaling that is strongly expressed in vessels during angiogenesis; however, the role of RGS5 in hypoxia has not been fully understood. Under hypoxic conditions, we found that the expression of RGS5, but not other RGS, was induced in human umbilical vein endothelial cells (HUVEC). RGS5 mRNA was increased when HUVEC were incubated with chemicals that stabilized hypoxia-inducible factor-1alpha (HIF-1alpha), whereas hypoxia-stimulated RGS5 promoter activity was absent in HIF-1beta(-/-) cells. Vascular endothelial growth factor (VEGF), which is regulated by HIF-1, did not appear to be involved in hypoxia-induced RGS5 expression; however, VEGF-mediated activation of p38 but not ERK1/2 was increased by RGS5. Overexpression of RGS5 in HUVEC exhibited a reduced growth rate without affecting the cell proliferation. Annexin V assay revealed that RGS5 induced apoptosis with significantly increased activation of caspase-3 and the Bax/Bcl-2 ratio. Small interfering RNA-specific for RGS5, caspase-3 inhibitor, and p38 inhibitor resulted in an attenuation of RGS5-stimulated apoptosis. Matrigel assay proved that RGS5 significantly impaired the angiogenic effect of VEGF and stimulated apoptosis in vivo. We concluded that RGS5 is a novel HIF-1-dependent, hypoxia-induced gene that is involved in the induction of endothelial apoptosis. Moreover, RGS5 antagonizes the angiogenic effect of VEGF by increasing the activation of p38 signaling, suggesting that RGS5 could be an important target for apoptotic therapy. PMID- 19564337 TI - Hemojuvelin-neogenin interaction is required for bone morphogenic protein-4 induced hepcidin expression. AB - Hemojuvelin (HJV) is a glycosylphosphatidylinositol-linked protein and binds both bone morphogenic proteins (BMPs) and neogenin. Cellular HJV acts as a BMP co receptor to enhance the transcription of hepcidin, a key iron regulatory hormone secreted predominantly by liver hepatocytes. In this study we characterized the role of neogenin in HJV-regulated hepcidin expression. Both HJV and neogenin were expressed in liver hepatocytes. Knockdown of neogenin decreased BMP4-induced hepcidin mRNA levels by 16-fold in HJV-expressing HepG2 cells but only by about 2 fold in cells transfected with either empty vector or G99V mutant HJV that does not bind BMPs. Further studies indicated that disruption of the HJV-neogenin interaction is responsible for a marked suppression of hepcidin expression. Moreover, in vivo studies showed that hepatic hepcidin mRNA could be significantly suppressed by blocking the interaction of HJV with full-length neogenin with a soluble fragment of neogenin in mice. Together, these results suggest that the HJV-neogenin interaction is required for the BMP-mediated induction of hepcidin expression when HJV is expressed. Combined with our previous studies, our results support that hepatic neogenin possesses two functions, mediation of cellular HJV release, and stimulation of HJV-enhanced hepcidin expression. PMID- 19564340 TI - MASP-1, a promiscuous complement protease: structure of its catalytic region reveals the basis of its broad specificity. AB - Mannose-binding lectin (MBL)-associated serine protease (MASP)-1 is an abundant component of the lectin pathway of complement. The related enzyme, MASP-2 is capable of activating the complement cascade alone. Though the concentration of MASP-1 far exceeds that of MASP-2, only a supporting role of MASP-1 has been identified regarding lectin pathway activation. Several non-complement substrates, like fibrinogen and factor XIII, have also been reported. MASP-1 belongs to the C1r/C1s/MASP family of modular serine proteases; however, its serine protease domain is evolutionary different. We have determined the crystal structure of the catalytic region of active MASP-1 and refined it to 2.55 A resolution. Unusual features of the structure are an internal salt bridge (similar to one in factor D) between the S1 Asp189 and Arg224, and a very long 60 loop. The functional and evolutionary differences between MASP-1 and the other members of the C1r/C1s/MASP family are reflected in the crystal structure. Structural comparison of the protease domains revealed that the substrate binding groove of MASP-1 is wide and resembles that of trypsin rather than early complement proteases explaining its relaxed specificity. Also, MASP-1's multifunctional behavior as both a complement and a coagulation enzyme is in accordance with our observation that antithrombin in the presence of heparin is a more potent inhibitor of MASP-1 than C1 inhibitor. Overall, MASP-1 behaves as a promiscuous protease. The structure shows that its substrate binding groove is accessible; however, its reactivity could be modulated by an unusually large 60 loop and an internal salt bridge involving the S1 Asp. PMID- 19564338 TI - Role of ADAMs in the ectodomain shedding and conformational conversion of the prion protein. AB - The cellular prion protein (PrP(C)) is essential for the pathogenesis and transmission of prion diseases. PrP(C) is bound to the plasma membrane via a glycosylphosphatidylinositol anchor, although a secreted, soluble form has also been identified. Previously we reported that PrP(C) is subject to ectodomain shedding from the membrane by zinc metalloproteinases with a similar inhibition profile to those involved in shedding the amyloid precursor protein. Here we have used gain-of-function (overexpression) and loss-of-function (small interfering RNA knockdown) experiments in cells to identify the ADAMs (a disintegrin and metalloproteinases) involved in the ectodomain shedding of PrP(C). These experiments revealed that ADAM9 and ADAM10, but not ADAM17, are involved in the shedding of PrP(C) and that ADAM9 exerts its effect on PrP(C) shedding via ADAM10. Using dominant negative, catalytically inactive mutants, we show that the catalytic activity of ADAM9 is required for its effect on ADAM10. Mass spectrometric analysis revealed that ADAM10, but not ADAM9, cleaved PrP between Gly(228) and Arg(229), three residues away from the site of glycosylphosphatidylinositol anchor attachment. The shedding of another membrane protein, the amyloid precursor protein beta-secretase BACE1, by ADAM9 is also mediated via ADAM10. Furthermore, we show that pharmacological inhibition of PrP(C) shedding or activation of both PrP(C) and PrP(Sc) shedding by ADAM10 overexpression in scrapie-infected neuroblastoma N2a cells does not alter the formation of proteinase K-resistant PrP(Sc). Collectively, these data indicate that although PrP(C) can be shed through the action of ADAM family members, modulation of PrP(C) or PrP(Sc) ectodomain shedding does not regulate prion conversion. PMID- 19564339 TI - Differential association of programmed death-1 and CD57 with ex vivo survival of CD8+ T cells in HIV infection. AB - Recent studies have revealed the critical role of programmed death-1 (PD-1) in exhaustion of HIV- and SIV-specific CD8(+) T cells. In this study, we show that high expression of PD-1 correlates with increased ex vivo spontaneous and CD95/Fas-induced apoptosis, particularly in the "effector-memory" CD8(+) T cell population from HIV(+) donors. High expression of PD-1 was linked to a proapoptotic phenotype characterized by low expression of Bcl-2 and IL7-R alpha, high expression of CD95/Fas and high mitochondrial mass. Expression of PD-1 and CD57 was differentially associated with the maturation status of CD8(+) T cells in HIV infection. CD57 was linked to higher apoptosis resistance, with cells expressing a PD-1(L)CD57(H) phenotype exhibiting lower levels of cell death. The majority of HIV-specific CD8(+) T cells were found to express a PD-1(H)CD57(L) or PD-1(H)CD57(H) phenotype. No correlation was found between PD-1 expression and ex vivo polyfunctionality of either HIV- or CMV-specific CD8(+) T cells. Contrary to CD57, high expression of PD-1 was characterized by translocation of PD-1 into the area of CD95/Fas-capping, an early necessary step of CD95/Fas-induced apoptosis. Thus, our data further support the role of PD-1 as a preapoptotic factor for CD8(+) T cells in HIV infection. PMID- 19564343 TI - VAMP8 is essential in anaphylatoxin-induced degranulation, TNF-alpha secretion, peritonitis, and systemic inflammation. AB - VAMP8, a member of the soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) family of fusion proteins, initially characterized in endosomal and endosomal-lysosomal fusion, may also function in regulated exocytosis. VAMP8 physiological function in inflammation has not been elucidated. In this paper, we show that deficiency of VAMP8 protects mice from anaphylatoxin (C5a)-induced neutropenia, peritonitis, and systemic inflammation. We show that, in vivo, VAMP8 deletion inhibits neutropenia and phagocyte recruitment. We also show that in macrophages, VAMP8 localizes on secretory granules and degranulation is inhibited in VAMP8-deficient macrophages. Moreover, VAMP8(-/-) mice show reduced systemic inflammation with inhibition of serum TNF alpha levels, whereas IL-1beta, IL-6, and MIP1alpha release are not affected. In wild-type macrophages, TNF-alpha colocalizes with VAMP8-positive vesicles, and in VAMP8-deficient macrophages, the TNF-alpha release is inhibited. Furthermore, VAMP8 regulates the release of TNF-alpha and beta-hexosaminidase triggered by fMLP, and VAMP8(-/-) mice are protected from fMLP-induced peritonitis. These data demonstrate that the VAMP8 vesicle-associated-SNARE is required for the proper trafficking of secretory lysosomal granules for exocytosis in macrophages and for the release of the potent proinflammatory cytokine, TNF-alpha. PMID- 19564341 TI - Regulation of dendritic cells and macrophages by an anti-apoptotic cell natural antibody that suppresses TLR responses and inhibits inflammatory arthritis. AB - Although natural Abs (NAbs) are present from birth, little is known about what drives their selection and whether they have housekeeping functions. The prototypic T15-NAb, first identified because of its protective role in infection, is representative of a special type of NAb response that specifically recognizes and forms complexes with apoptotic cells and which promotes cell-corpse engulfment by phagocytes. We now show that this T15-NAb IgM-mediated clearance process is dependent on the recruitment of C1q and mannose-binding lectin, which have known immune modulatory activities that also provide "eat me" signals for enhancing phagocytosis. Further investigation revealed that the addition of T15 NAb significantly suppressed in vitro LPS-induced TNF-alpha and IL-6 secretion by the macrophage-like cell line, RAW264.7, as well as TLR3-, TLR4-, TLR7-, and TLR9 induced maturation and secretion of a range of proinflammatory cytokines and chemokines by bone marrow-derived conventional dendritic cells. Significantly, high doses of this B-1 cell produced NAb also suppressed in vivo TLR-induced proinflammatory responses. Although infusions of apoptotic cells also suppressed such in vivo inflammatory responses and this effect was associated with the induction of high levels of IgM antiapoptotic cell Abs, apoptotic cell treatment was not effective at suppressing such TLR responses in B cell-deficient mice. Moreover, infusions of T15-NAb also efficiently inhibited both collagen-induced arthritis and anti-collagen II Ab-mediated arthritis. These studies identify and characterize a previously unknown regulatory circuit by which a NAb product of innate-like B cells aids homeostasis by control of fundamental inflammatory pathways. PMID- 19564342 TI - FOXP3 inhibits activation-induced NFAT2 expression in T cells thereby limiting effector cytokine expression. AB - The forkhead DNA-binding protein FOXP3 is critical for the development and suppressive function of CD4(+)CD25(+) regulatory T cells (T(REG)), which play a key role in maintaining self-tolerance. Functionally, FOXP3 is capable of repressing transcription of cytokine genes regulated by NFAT. Various mechanisms have been proposed by which FOXP3 mediates these effects. Using novel cell lines that inducibly express either wild-type or mutant FOXP3, we have identified NFAT2 as an early target of FOXP3-mediated transcriptional repression. NFAT2 is typically expressed at low levels in resting T cells, but is up-regulated by NFAT1 upon cellular activation. We demonstrate that transcription from the NFAT2 promoter is significantly suppressed by FOXP3, and NFAT2 protein expression is markedly diminished in activated CD4(+)CD25(+)FOXP3(+) T(REG) compared with CD4(+)CD25(-)FOXP3(-) T cells. Chromatin immunoprecipitation experiments indicate that FOXP3 competes with NFAT1 for binding to the endogenous NFAT2 promoter. This antagonism of NFAT2 activity by FOXP3 is important for the anergic phenotype of T(REG), as ectopic expression of NFAT2 from a retroviral LTR partially restores expression of IL-2 in FOXP3(+) T(REG). These data suggest that FOXP3 functions not only to suppress the first wave of NFAT-mediated transcriptional responses, but may also affect sustained NFAT-mediated inflammatory gene expression through suppression of inducible NFAT2 transcription. PMID- 19564345 TI - The induction of IL-10 by zymosan in dendritic cells depends on CREB activation by the coactivators CREB-binding protein and TORC2 and autocrine PGE2. AB - Stimulation of human monocyte-derived dendritic cells with the yeast extract zymosan is characterized by a predominant production of IL-10 and a strong induction of cyclooxygenase-2, but the molecular mechanisms underlying this response are only partially understood. To address this issue, the activation of transcription factors that may bind to the il10 proximal promoter was studied. Binding activity to Sp1, Sp3, NF-Y, and cAMP response element (CRE) sites was detected in the nuclear extracts of dendritic cells; however these binding activities were not influenced by zymosan. No binding activity to Stat1, Stat3, and c/EBP sites was detected. Notably, zymosan activated kappaB-binding activity, but inhibition of NF-kappaB was associated with enhanced IL-10 production. In sharp contrast, treatments acting on CREB (CRE binding protein), including 8-Br cAMP, PGE(2), and inhibitors of PKA, COX, and glycogen-synthase kinase-3beta showed a direct correlation between CREB activation and IL-10 production. Zymosan induced binding of both P-CREB and CREB-binding protein (CBP) to the il10 promoter as judged from chromatin immunoprecipitation assays, whereas negative results were obtained with Ab reactive to Sp1, Sp3, c-Maf, and NF-Y. Zymosan also induced nuclear translocation of the CREB coactivator transducer of regulated CREB activity 2 (TORC2) and interaction of TORC2 with P-CREB coincidental with the association of CREB to the il10 promoter. Altogether, our data show that zymosan induces il10 transcription by a CRE-dependent mechanism that involves autocrine secretion of PGE(2) and a network of interactions of PKA, MAP/ERK, glycogen-synthase kinase-3beta, and calcineurin, which regulate CREB transcriptional activity by binding the coactivators CBP and TORC2 and inhibiting CBP interaction with other transcription factors. PMID- 19564344 TI - Reduced cytotoxic function of effector CD8+ T cells is responsible for indoleamine 2,3-dioxygenase-dependent immune suppression. AB - Indoleamine 2,3-dioxygenase (IDO), a potent immunosuppressive enzyme, contributes to tumoral escape, immune tolerance, and protection against allograft injury. In this paper, we report that inhibition of CD8(+) T cell-mediated cytotoxic function is an important mechanism behind IDO's immune-modulating property. The experimental rat lung allograft proved attractive for evaluating effector CD8(+) T cells. Enhanced IDO activity achieved by using a lung-tissue-targeted nonviral human IDO gene transfer approach reduced, but did not eliminate, infiltrating CD8(+) T cells. Although CD8(+) T cells existed in the IDO-high lung allografts, CD8(+) T cells remained viable and could proliferate for an extended period. However, cells lost their ability to attack allogeneic donor lung cells in vivo and allogeneic target cells in vitro. The impaired cytotoxic function seen in the IDO-treated CD8(+) T cells was accompanied by defects in production of granule cytotoxic proteins, including perforin and granzyme A and B. Furthermore, we discovered that IDO leads to an impaired bioenergetic condition in active CD8(+) T cells via selective inhibition of complex I in the mitochondrial electron transfer chain. These intriguing findings provide a base for establishing a novel mode of IDO's immune-suppressing action. Additionally, donor lung IDO delivery, a direct and/or leukocyte passenger effect, impaired CD8(+) effector cell function. PMID- 19564346 TI - Ablation and regeneration of tolerance-inducing medullary thymic epithelial cells after cyclosporine, cyclophosphamide, and dexamethasone treatment. AB - Immunosuppressive drugs and cytotoxic chemotherapy agents are designed to kill or suppress autoreactive, alloaggressive, or hyperinflammatory T cells, or disseminated malignancies. However, they also cause severe immunological side effects ranging from interrupted thymopoiesis and general immunodeficiency to, paradoxically, autoimmunity. Consistent with the cross-talk between thymocytes and stromal cells, we now show that these common therapeutic agents have major effects on murine thymic epithelial cells (TEC), crucially required to rebuild immunity posttreatment. We show that the immunosuppressant cyclosporine A, which has been linked to a thymus-dependent autoimmune syndrome in some patients, causes extensive loss of autoimmune regulator (Aire(+)) tolerance-inducing MHC class II(high) medullary TEC (mTEC(high)). Post-cyclosporine A, Aire expression was restored within 7 days. Full recovery of the mTEC(high) subset occurred within 10 days and was linked to a decrease in a relatively resistant MHC class II(low) mTEC subset (mTEC(low)), consistent with a previously described precursor product relationship. Cyclophosphamide and dexamethasone caused more extensive ablation of thymocytes and stromal cells but again severely depleted tolerance inducing mTEC(high). Together, these data show that Aire(+) mTECs are highly sensitive to damage and that mTEC regeneration follows a conserved pattern regardless of the treatment regimen used. PMID- 19564350 TI - STAT3 links IL-22 signaling in intestinal epithelial cells to mucosal wound healing. AB - Signal transducer and activator of transcription (STAT) 3 is a pleiotropic transcription factor with important functions in cytokine signaling in a variety of tissues. However, the role of STAT3 in the intestinal epithelium is not well understood. We demonstrate that development of colonic inflammation is associated with the induction of STAT3 activity in intestinal epithelial cells (IECs). Studies in genetically engineered mice showed that epithelial STAT3 activation in dextran sodium sulfate colitis is dependent on interleukin (IL)-22 rather than IL 6. IL-22 was secreted by colonic CD11c(+) cells in response to Toll-like receptor stimulation. Conditional knockout mice with an IEC-specific deletion of STAT3 activity were highly susceptible to experimental colitis, indicating that epithelial STAT3 regulates gut homeostasis. STAT3(IEC-KO) mice, upon induction of colitis, showed a striking defect of epithelial restitution. Gene chip analysis indicated that STAT3 regulates the cellular stress response, apoptosis, and pathways associated with wound healing in IECs. Consistently, both IL-22 and epithelial STAT3 were found to be important in wound-healing experiments in vivo. In summary, our data suggest that intestinal epithelial STAT3 activation regulates immune homeostasis in the gut by promoting IL-22-dependent mucosal wound healing. PMID- 19564349 TI - Dendritic cells require a systemic type I interferon response to mature and induce CD4+ Th1 immunity with poly IC as adjuvant. AB - Relative to several other toll-like receptor (TLR) agonists, we found polyinosinic:polycytidylic acid (poly IC) to be the most effective adjuvant for Th1 CD4(+) T cell responses to a dendritic cell (DC)-targeted HIV gag protein vaccine in mice. To identify mechanisms for adjuvant action in the intact animal and the polyclonal T cell repertoire, we found poly IC to be the most effective inducer of type I interferon (IFN), which was produced by DEC-205(+) DCs, monocytes, and stromal cells. Antibody blocking or deletion of type I IFN receptor showed that IFN was essential for DC maturation and development of CD4(+) immunity. The IFN-AR receptor was directly required for DCs to respond to poly IC. STAT 1 was also essential, in keeping with the type I IFN requirement, but not type II IFN or IL-12 p40. Induction of type I IFN was mda5 dependent, but DCs additionally used TLR3. In bone marrow chimeras, radioresistant and, likely, nonhematopoietic cells were the main source of IFN, but mda5 was required in both marrow-derived and radioresistant host cells for adaptive responses. Therefore, the adjuvant action of poly IC requires a widespread innate type I IFN response that directly links antigen presentation by DCs to adaptive immunity. PMID- 19564351 TI - IL-17 can promote tumor growth through an IL-6-Stat3 signaling pathway. AB - Although the Th17 subset and its signature cytokine, interleukin (IL)-17A (IL 17), are implicated in certain autoimmune diseases, their role in cancer remains to be further explored. IL-17 has been shown to be elevated in several types of cancer, but how it might contribute to tumor growth is still unclear. We show that growth of B16 melanoma and MB49 bladder carcinoma is reduced in IL-17(-/-) mice but drastically accelerated in IFN-gamma(-/-) mice, contributed to by elevated intratumoral IL-17, indicating a role of IL-17 in promoting tumor growth. Adoptive transfer studies and analysis of the tumor microenvironment suggest that CD4(+) T cells are the predominant source of IL-17. Enhancement of tumor growth by IL-17 involves direct effects on tumor cells and tumor-associated stromal cells, which bear IL-17 receptors. IL-17 induces IL-6 production, which in turn activates oncogenic signal transducer and activator of transcription (Stat) 3, up-regulating prosurvival and proangiogenic genes. The Th17 response can thus promote tumor growth, in part via an IL-6-Stat3 pathway. PMID- 19564352 TI - A mutation in Irak2c identifies IRAK-2 as a central component of the TLR regulatory network of wild-derived mice. AB - In a phenotypic screen of the wild-derived mouse strain MOLF/Ei, we describe an earlier and more potent toll-like receptor (TLR)-mediated induction of IL-6 transcription compared with the classical inbred strain C57BL/6J. The phenotype correlated with increased activity of the IkappaB kinase axis as well as p38, but not extracellular signal-regulated kinase or c-Jun N-terminal kinase, mitogen activated protein kinase (MAPK) phosphorylation. The trait was mapped to the Why1 locus, which contains Irak2, a gene previously implicated as sustaining the late phase of TLR responses. In the MOLF/Ei TLR signaling network, IRAK-2 promotes early nuclear factor kappaB (NF-kappaB) activity and is essential for the activation of p38 MAPK. We identify a deletion in the MOLF/Ei promoter of the inhibitory Irak2c gene, leading to an increased ratio of pro- to antiinflammatory IRAK-2 isoforms. These findings demonstrate that IRAK-2 is an essential component of the early TLR response in MOLF/Ei mice and show a distinct pathway of p38 and NF-kappaB activation in this model organism. In addition, they demonstrate that studies in evolutionarily divergent model organisms are essential to complete dissection of signal transduction pathways. PMID- 19564353 TI - Human naive and memory CD4+ T cell repertoires specific for naturally processed antigens analyzed using libraries of amplified T cells. AB - The enormous diversity of the naive T cell repertoire is instrumental in generating an immune response to virtually any foreign antigen that can be processed into peptides that bind to MHC molecules. The low frequency of antigen specific naive T cells, their high activation threshold, and the constrains of antigen-processing and presentation have hampered analysis of naive repertoires to complex protein antigens. In this study, libraries of polyclonally expanded naive T cells were used to determine frequency and antigen dose-response of human naive CD4(+) T cells specific for a variety of antigens and to isolate antigen specific T cell clones. In the naive repertoire, T cells specific for primary antigens, such as KLH and Bacillus anthracis protective antigen, and for recall antigens, such as tetanus toxoid, cytomegalovirus, and Mycobacterium tuberculosis purified protein derivative, were detected at frequencies ranging from 5 to 170 cells per 10(6) naive T cells. Antigen concentrations required for half-maximal response (EC50) varied over several orders of magnitude for different naive T cells. In contrast, in the memory repertoire, T cells specific for primary antigens were not detected, whereas T cells specific for recall antigens were detected at high frequencies and displayed EC50 values in the low range of antigen concentrations. The method described may find applications for evaluation of vaccine candidates, for testing antigenicity of therapeutic proteins, drugs, and chemicals, and for generation of antigen-specific T cell clones for adoptive cellular immunotherapy. PMID- 19564356 TI - Proceedings of Workshop on Emergency Preparedness for Vulnerable Population Groups. March 2-3, 2009. Ottawa, Ontario, Canada. PMID- 19564354 TI - Regulation of TLR7/9 responses in plasmacytoid dendritic cells by BST2 and ILT7 receptor interaction. AB - Plasmacytoid dendritic cells (pDCs) produce copious type I interferon (IFN) upon sensing nucleic acids through Toll-like receptor (TLR) 7 and TLR9. Uncontrolled pDC activation and IFN production are implicated in lymphopenia and autoimmune diseases; therefore, a mechanism controlling pDC IFN production is essential. Human pDCs specifically express an orphan receptor, immunoglobulin-like transcript 7 (ILT7). Here, we discovered an ILT7 ligand expressed by human cell lines and identified it as bone marrow stromal cell antigen 2 (BST2; CD317). BST2 directly binds to purified ILT7 protein, initiates signaling via the ILT7 FcepsilonRIgamma complex, and strongly inhibits production of IFN and proinflammatory cytokines by pDCs. Readily induced by IFN and other proinflammatory cytokines, BST2 may modulate the human pDC's IFN responses through ILT7 in a negative feedback fashion. PMID- 19564355 TI - The thymic medulla: a unique microenvironment for intercellular self-antigen transfer. AB - Central tolerance is shaped by the array of self-antigens expressed and presented by various types of thymic antigen-presenting cells (APCs). Depending on the overall signal quality and/or quantity delivered in these interactions, self reactive thymocytes either apoptose or commit to the T regulatory cell lineage. The cellular and molecular complexity underlying these events has only recently been appreciated. We analyzed the ex vivo presentation of ubiquitous or tissue restricted self-antigens by medullary thymic epithelial cells (mTECs) and thymic dendritic cells (DCs), the two major APC types present in the medulla. We found that the ubiquitously expressed nuclear neo-self-antigen ovalbumin (OVA) was efficiently presented via major histocompatibility complex class II by mTECs and thymic DCs. However, presentation by DCs was highly dependent on antigen expression by TECs, and hemopoietic cells did not substitute for this antigen source. Accordingly, efficient deletion of OVA-specific T cells correlated with OVA expression by TECs. Notably, OVA was only presented by thymic but not peripheral DCs. We further demonstrate that thymic DCs are constitutively provided in situ with cytosolic as well as membrane-bound mTEC-derived proteins. The subset of DCs displaying transferred proteins was enriched in activated DCs, with these cells being most efficient in presenting TEC-derived antigens. These data provide evidence for a unique, constitutive, and unidirectional transfer of self-antigens within the thymic microenvironment, thus broadening the cellular base for tolerance induction toward promiscuously expressed tissue antigens. PMID- 19564357 TI - Profound antiviral effect of oral administration of MIV-210 on chronic hepadnaviral infection in a woodchuck model of hepatitis B. AB - MIV-210 is a prodrug of 3'-fluoro-2',3'-dideoxyguanosine with high oral bioavailability in humans and potent activity against hepatitis B virus (HBV). Woodchucks infected with woodchuck hepatitis virus (WHV) represent an accurate model of HBV infection that is utilized for evaluation of the efficacy and safety of novel anti-HBV agents. Oral administration of MIV-210 at 20 or 60 mg/kg of body weight/day induced a rapid virological response in chronically infected woodchucks, reducing serum WHV DNA levels by 4.75 log10 and 5.72 log10, respectively, in 2 weeks. A progressive decline in WHV viremia occurred throughout the 10-week therapy, giving final reductions of 7.23 log10 and 7.68 log10 in the 20- and 60-mg/kg/day groups, respectively. Further, a daily dose of 10 mg/kg decreased the serum WHV load 400-fold after 4 weeks of treatment, and a dose of 5 mg/kg/day was sufficient to maintain this antiviral effect during the following 6-week period. MIV-210 at 20 or 60 mg/kg/day reduced the liver WHV DNA load 200- to 2,500-fold from pretreatment levels and, importantly, led to a 2.0 log10 drop in the hepatic content of WHV covalently closed circular DNA. The treatment with 60 mg/kg/day was well tolerated. Liver biopsy specimens obtained after the 10-week treatment with 20 or 60 mg/kg/day and after the 10-week follow up showed hepatocyte and mitochondrial ultrastructures comparable to those in the placebo-treated group. It was concluded that MIV-210 is highly effective against chronic WHV infection. These findings, together with the previously demonstrated inhibitory activity of MIV-210 against lamivudine-, adefovir-, and entecavir resistant HBV variants, make MIV-210 a highly valuable candidate for further testing as an agent against chronic hepatitis B. PMID- 19564358 TI - Tuberculosis and trimethoprim-sulfamethoxazole. AB - The sulfonamides were the first drugs with antituberculous effects. Their use was abandoned and basically forgotten with the advent of streptomycin and isoniazid combination treatment. There is a widespread belief, apparently based on testing a single isolate on questionable media, that Mycobacterium tuberculosis is resistant to trimethoprim-sulfamethoxazole (TMP-SMX). We saw a complex immunocompromised patient with tuberculosis who was initially treated with TMP SMX without antituberculous drugs and defervesced on this treatment. An isolate of M. tuberculosis from this patient was found to be sensitive to TMP-SMX. We examined how frequently M. tuberculosis is sensitive to TMP-SMX. Isolates were tested for susceptibility to TMP-SMX on supplemented Middlebrook 7H10 plates. We found that 43 of 44 (98%) isolates of M. tuberculosis were susceptible to the combination of < or = 1 microg/ml of TMP and 19 microg/ml of SMX (< or = 1/19 microg/ml). Thus, the vast majority of our M. tuberculosis isolates were susceptible to TMP-SMX at an MIC similar to that for Mycobacterium kansasii, Mycobacterium marinum, and sensitive rapidly growing mycobacteria, organisms successfully treated with TMP-SMX as part of the treatment regimen. It is possible that TMP-SMX may be useful in treating patients with multiple-drug resistant and extended drug-resistant tuberculosis. We feel that a clinical trial looking at the effectiveness of TMP-SMX as an antituberculous drug is worthwhile. PMID- 19564359 TI - Antiprotozoal activity of 1-phenethyl-4-aminopiperidine derivatives. AB - A series of 44 4-aminopiperidine derivatives was screened in vitro against four protozoan parasites (Trypanosoma brucei rhodesiense, Trypanosoma cruzi, Leishmania donovani, and Plasmodium falciparum). This screening identified 29 molecules selectively active against bloodstream-form T. b. rhodesiense trypomastigotes, with 50% inhibitory concentrations (IC50) ranging from 0.12 to 10 microM, and 33 compounds active against the chloroquine- and pyrimethamine resistant K1 strain of P. falciparum (IC50 range, 0.17 to 5 microM). In addition, seven compounds displayed activity against intracellular T. cruzi amastigotes in the same range as the reference drug benznidazole (IC50, 1.97 microM) but were also cytotoxic to L-6 cells, showing little selectivity for T. cruzi. None of the molecules tested showed interesting antileishmanial activity against axenic amastigotes of L. donovani. To our knowledge, this is the first report of the antitrypanosomal activity of molecules bearing the 4-aminopiperidine skeleton. PMID- 19564360 TI - Probing the differential interactions of quinazolinedione PD 0305970 and quinolones with gyrase and topoisomerase IV. AB - Quinazoline-2,4-diones, such as PD 0305970, are new DNA gyrase and topoisomerase IV (topo IV) inhibitors with potent activity against gram-positive pathogens, including quinolone-resistant isolates. The mechanistic basis of dione activity vis-a-vis quinolones is not understood. We present evidence for Streptococcus pneumoniae gyrase and topo IV that PD 0305970 and quinolones interact differently with the enzyme breakage-reunion and Toprim domains, DNA, and Mg2+-four components that are juxtaposed in the topoisomerase cleavage complex to effect DNA scission. First, PD 0305970 targets primarily gyrase in Streptococcus pneumoniae. However, unlike quinolones, which select predominantly for gyrA (or topo IV parC) mutations in the breakage-reunion domain, unusually the dione selected for novel mutants with alterations that map to a region of the Toprim domain of GyrB (R456H and E474A or E474D) or ParE (D435H and E475A). This "dione resistance-determining region" overlaps the GyrB quinolone resistance-determining region and the region that binds essential Mg2+ ions, each function involving conserved EGDSA and PLRGK motifs. Second, dione-resistant gyrase and topo IV were inhibited by ciprofloxacin, whereas quinolone-resistant enzymes (GyrA S81F and ParC S79F) remained susceptible to PD 0305970. Third, dione-promoted DNA cleavage by gyrase occurred at a distinct repertoire of sites, implying that structural differences with quinolones are sensed at the DNA level. Fourth, unlike the situation with quinolones, the Mg2+ chelator EDTA did not reverse dione-induced gyrase cleavage nor did the dione promote Mg2+-dependent DNA unwinding. It appears that PD 0305970 interacts uniquely to stabilize the cleavage complex of gyrase/topo IV perhaps via an altered orientation directed by the bidentate 3 amino-2,4-dione moiety. PMID- 19564361 TI - Population pharmacokinetics of linezolid in adults with pulmonary tuberculosis. AB - Nineteen adults with pulmonary tuberculosis received linezolid (600 mg) once or twice daily in an early bactericidal activity trial. A one-compartment population model produced median values for the absorption rate constant, volume of distribution, and elimination rate constant of 1.5 h(-1), 29.6 liters, and 0.25 h(-1) (once daily) and 2.7 h(-1), 32.1 liters, and 0.15 h(-1) (twice daily). Linezolid administered twice daily produced higher values for free drug area under the concentration-time curve (AUC)/MIC and time above MIC. Both regimens achieved free AUC/MIC ratios > 100. Median times above the MIC for free drug were 100% (twice daily) and 63% (once daily). PMID- 19564362 TI - Efficacy of iclaprim against wild-type and thymidine kinase-deficient methicillin resistant Staphylococcus aureus isolates in an in vitro fibrin clot model. AB - Iclaprim is a novel diaminopyrimidine antibiotic that is active against methicillin-resistant Staphylococcus aureus (MRSA). However, it is known that the activity of diaminopyrimidines against S. aureus is antagonized by thymidine through uptake and conversion to thymidylate by thymidine kinase. Unlike with humans, for whom thymidine levels are low, thymidine levels in rodents are high, thus precluding the accurate evaluation of iclaprim efficacy in animal models. We have studied the bactericidal activity of iclaprim against an isogenic pair of MRSA isolates, the wild-type parent AW6 and its thymidine kinase-deficient mutant AH1252, in an in vitro fibrin clot model. Clots, which were aimed at mimicking vegetation structure, were made from human or rat plasma containing either the parent AW6 or the mutant AH1252, and they were exposed to homologous serum supplemented with iclaprim (3.5 microg/ml), trimethoprim-sulfamethoxazole (TMP SMX; 8/40 microg/ml), vancomycin (40 microg/ml), or saline, each of which was added one time for 48 h. In rat clots, iclaprim and TMP-SMX were bacteriostatic against the parent, AW6. In contrast, they were bactericidal (> or = 3 log10 CFU/clot killing of the original inoculum) against the mutant AH1252. Vancomycin was the most active drug against AW6 (P < 0.05), but it showed an activity similar those of iclaprim and TMP-SMX against AH1252. In human clots, iclaprim was bactericidal against both AW6 and AH1252 strains and was as effective as TMP SMX and vancomycin (P > 0.05). Future studies of animals using simulated human kinetics of iclaprim and thymidine kinase-deficient MRSA, which eliminate the thymidine-induced confounding effect, are warranted to support the use of iclaprim in the treatment of severe MRSA infections in humans. PMID- 19564363 TI - Activities of daptomycin and vancomycin alone and in combination with rifampin and gentamicin against biofilm-forming methicillin-resistant Staphylococcus aureus isolates in an experimental model of endocarditis. AB - The findings of clinical and in vitro research support the theory that infective endocarditis (IE)-causing bacteria form biofilms and that biofilms negatively affect treatment outcomes. The purpose of the present study was to quantify the biofilm formation of methicillin (meticillin)-resistant Staphylococcus aureus (MRSA) isolates obtained from patients with IE and to evaluate the in vitro activities of daptomycin and vancomycin alone and in combination with rifampin (rifampicin) or gentamicin while monitoring the isolates for the development of resistance. A high-inoculum, stationary-phase infection model of IE was used to simulate the pharmacokinetics in humans of daptomycin at 6 mg/kg of body weight/day, vancomycin at 1.25 g every 12 h (q12h) alone and in combination with rifampin at 300 mg every 8 h, and gentamicin at 1.3 mg/kg q12h. Two randomly selected clinical MRSA isolates were obtained from patients with IE; both MRSA isolates quantitatively produced biofilms. The time to bactericidal activity in the presence of daptomycin was isolate dependent but was achieved by 24 h for both MRSA isolates. Vancomycin did not achieve bactericidal activity throughout the experiment. At 24, 48, and 72 h, daptomycin-containing regimens had significantly more activity (greater declines in the mean number of CFU/g) than any of the vancomycin-containing regimens (P = 0.03). Rifampin and gentamicin antagonized or delayed the bactericidal activity of daptomycin (against MRSA B346846 for rifampin and against both isolates for gentamicin) in the first 24 h. Increases in the daptomycin and vancomycin MICs were not observed. We conclude that in an IE model of biofilm-forming MRSA, daptomycin monotherapy has better in vitro activity than daptomycin in combination with rifampin or gentamicin or any vancomycin-containing regimen studied within the first 24 h. Further investigations are needed to understand the initial delay in bactericidal activity observed when gentamicin or rifampin is combined with daptomycin. PMID- 19564364 TI - Effect of hemagglutinin-neuraminidase inhibitors BCX 2798 and BCX 2855 on growth and pathogenicity of Sendai/human parainfluenza type 3 chimera virus in mice. AB - Human parainfluenza virus type 3 (hPIV-3) is a major respiratory tract pathogen that affects young children, but no vaccines or antiviral drugs against it have yet been developed. We developed a mouse model to evaluate the efficacies of the novel parainfluenza virus hemagglutinin-neuraminidase (HN) inhibitors BCX 2798 and BCX 2855 against a recombinant Sendai virus (rSeV) in which the fusion (F) and HN surface glycoproteins (FHN) were replaced by those of hPIV-3 [rSeV(hPIV 3FHN)]. In the prophylaxis model, 129X1/SvJ mice were infected with a 90% or 20% lethal dose of the virus and were treated intranasally for 5 days with 10 mg/kg of body weight/day of either compound starting 4 h before infection. Prophylactic treatment of the mice with either compound did not prevent their death in a 90% lethality model of rSeV(hPIV-3FHN) infection. However, it significantly reduced the lung virus titers, the amount of weight lost, and the rate of mortality in mice infected with a 20% lethal virus dose. In the therapy model, mice were infected with a nonlethal dose of the virus (100 PFU/mouse) and were treated intranasally with 1 or 10 mg/kg/day of either compound for 5 days starting at 24 or 48 h postinfection. Treatment of the mice with either compound significantly reduced the virus titer in the lungs, subsequently causing a reduction in the number of immune cells and the levels of cytokines in the bronchoalveolar lavage fluid and histopathologic changes in the airways. Our results indicate that BCX 2798 and BCX 2855 are effective inhibitors of hPIV-3 HN in our mouse model and may be promising candidates for the prophylaxis and treatment of hPIV-3 infection in humans. PMID- 19564365 TI - Cellular accumulation and pharmacodynamic evaluation of the intracellular activity of CEM-101, a novel fluoroketolide, against Staphylococcus aureus, Listeria monocytogenes, and Legionella pneumophila in human THP-1 macrophages. AB - CEM-101 is a novel fluoroketolide with lower MICs than those of telithromycin and macrolides. Our aim was to assess the cellular accumulation and intracellular activity of CEM-101 using models developed for analyzing the pharmacokinetics and pharmacological properties of antibiotics against phagocytized bacteria. We used THP-1 macrophages and Staphylococcus aureus (ATCC 25923 [methicillin (meticillin) sensitive]), Listeria monocytogenes (strain EGD), and Legionella pneumophila (ATCC 33153). CEM-101 reached cellular-to-extracellular-concentration ratios of about 350 within 24 h (versus approximately 20, 30, and 160 for telithromycin, clarithromycin, and azithromycin, respectively). This intracellular accumulation was suppressed by incubation at a pH of < or = 6 and by monensin (proton ionophore) and was unaffected by verapamil (P-glycoprotein inhibitor; twofold accumulation increase for azithromycin) or gemfibrozil. While keeping with the general properties of the macrolide antibiotics in terms of maximal efficacy (Emax; approximately 1-log10-CFU decrease compared to the postphagocytosis inoculum after a 24-h incubation), CEM-101 showed significantly greater potency against phagocytized S. aureus than telithromycin, clarithromycin, and azithromycin (for which the 50% effective concentration [EC50] and static concentrations were about 3-, 6-, and 15-fold lower, respectively). CEM-101 was also about 50-fold and 100-fold more potent than azithromycin against phagocytized L. monocytogenes and L. pneumophila, respectively. These differences in EC50s and static concentrations between drugs were minimized when data were expressed as multiples of the MIC, demonstrating the critical role of intrinsic drug activity (MIC) in eliciting the antibacterial intracellular effects, whereas accumulation per se was unimportant. CEM-101 should show enhanced in vivo potency if used at doses similar to those of the comparators tested here. PMID- 19564367 TI - Overexpression of the naturally occurring blaOXA-51 gene in Acinetobacter baumannii mediated by novel insertion sequence ISAba9. PMID- 19564366 TI - Population pharmacokinetics of lumefantrine in pregnant women treated with artemether-lumefantrine for uncomplicated Plasmodium falciparum malaria. AB - Artemether-lumefantrine has become one of the most widely used antimalarial drugs in the world. The objective of this study was to determine the population pharmacokinetic properties of lumefantrine in pregnant women with uncomplicated multidrug-resistant Plasmodium falciparum malaria on the northwestern border of Thailand. Burmese and Karen women (n = 103) with P. falciparum malaria and in the second and third trimesters of pregnancy were treated with artemether lumefantrine (80/480 mg) twice daily for 3 days. All patients provided five capillary plasma samples for drug quantification, and the collection times were randomly distributed over 14 days. The concentration-time profiles of lumefantrine were assessed by nonlinear mixed-effects modeling. The treatment failure rate (PCR-confirmed recrudescent infections at delivery) was high; 16.5% (95% confidence interval, 9.9 to 25.1). The population pharmacokinetics of lumefantrine were described well by a two-compartment open model with first-order absorption and elimination. The final model included interindividual variability in all pharmacokinetic parameters and a linear covariate relationship between the estimated gestational age and the central volume of distribution. A high proportion of all women (40%, 41/103) had day 7 capillary plasma concentrations of <355 ng/ml (which corresponds to approximately <280 ng/ml in venous plasma), a threshold previously associated with an increased risk of therapeutic failure in nonpregnant patients in this area. Predictive modeling suggests that a twice daily regimen given for 5 days would be preferable in later pregnancy. In conclusion, altered pharmacokinetic properties of lumefantrine contribute to the high rates of failure of artemether-lumefantrine treatment in later pregnancy. Dose optimization is urgently needed. PMID- 19564368 TI - Competitive inhibition of renal tubular secretion of gemifloxacin by probenecid. AB - Probenecid interacts with transport processes of drugs at several sites in the body. For most quinolones, renal clearance is reduced by concomitant administration of probenecid. The interaction between gemifloxacin and probenecid has not yet been studied. We studied the extent, time course, site(s), and mechanism of this interaction. Seventeen healthy volunteers participated in a randomized, two-way crossover study. Subjects received 320 mg gemifloxacin as an oral tablet without and with 4.5 g probenecid divided in eight oral doses. Drug concentrations in plasma and urine were analyzed by liquid chromatography-tandem mass spectrometry. WinNonlin was used for noncompartmental analysis, compartmental modeling, and statistics, and NONMEM was used for visual predictive checks. Concomitant administration of probenecid increased plasma gemifloxacin concentrations and amounts excreted in urine compared to baseline amounts. Data are average estimates (percent coefficients of variation). Modeling showed a competitive inhibition of the renal tubular secretion of gemifloxacin by probenecid as the most likely mechanism of the interaction. The estimated K(m) and Vmax for the saturable part of renal elimination were 9.16 mg/liter (20%) and 113 mg/h (21%), respectively. Based on the molar ratio, the affinity for the renal transporter was 10-fold higher for gemifloxacin than for probenecid. Since probenecid reached an approximately 200-times-higher area under the molar concentration-time curve from 0 to 24 h than gemifloxacin, probenecid inhibited the active tubular secretion of gemifloxacin. Probenecid also reduced the nonrenal clearance of gemifloxacin from 25.2 (26%) to 21.0 (23%) liters/h. Probenecid inhibited the renal tubular secretion of gemifloxacin, most likely by a competitive mechanism, and slightly decreased nonrenal clearance of gemifloxacin. PMID- 19564369 TI - Photodynamic therapy for Acinetobacter baumannii burn infections in mice. AB - Multidrug-resistant Acinetobacter baumannii infections represent a growing problem, especially in traumatic wounds and burns suffered by military personnel injured in Middle Eastern conflicts. Effective treatment with traditional antibiotics can be extremely difficult, and new antimicrobial approaches are being investigated. One of these alternatives to antimicrobials could be the combination of nontoxic photosensitizers (PSs) and visible light, known as photodynamic therapy (PDT). We report on the establishment of a new mouse model of full-thickness thermal burns infected with a bioluminescent derivative of a clinical Iraqi isolate of A. baumannii and its PDT treatment by topical application of a PS produced by the covalent conjugation of chlorin(e6) to polyethylenimine, followed by illumination of the burn surface with red light. Application of 10(8) A. baumannii cells to the surface of 10-s burns made on the dorsal surface of shaved female BALB/c mice led to chronic infections that lasted, on average, 22 days and that were characterized by a remarkably stable bacterial bioluminescence. PDT carried out on day 0 soon after application of the bacteria gave over 3 log units of loss of bacterial luminescence in a light exposure-dependent manner, while PDT carried out on day 1 and day 2 gave an approximately 1.7-log reduction. The application of PS dissolved in 10% or 20% dimethyl sulfoxide without light gave only a modest reduction in the bacterial luminescence from mouse burns. Some bacterial regrowth in the treated burn was observed but was generally modest. It was also found that PDT did not lead to the inhibition of wound healing. The data suggest that PDT may be an effective new treatment for multidrug-resistant localized A. baumannii infections. PMID- 19564370 TI - Candida albicans and Staphylococcus aureus form polymicrobial biofilms: effects on antimicrobial resistance. AB - Candida albicans readily forms biofilms on the surface on indwelling medical devices, and these biofilms serve as a source of local and systemic infections. It is estimated that 27% of nosocomial C. albicans bloodstream infections are polymicrobial, with Staphylococcus aureus as the third most common organism isolated in conjunction with C. albicans. We tested whether S. aureus and C. albicans are able to form a polymicrobial biofilm. Although S. aureus formed poor monoculture biofilms in serum, it formed a substantial polymicrobial biofilm in the presence of C. albicans. In terms of architecture, S. aureus formed microcolonies on the surface of the biofilm, with C. albicans serving as the underlying scaffolding. In addition, S. aureus matrix staining revealed a different phenotype in polymicrobial versus monomicrobial biofilms, suggesting that S. aureus may become coated in the matrix secreted by C. albicans. S. aureus resistance to vancomycin was enhanced within the polymicrobial biofilm, required viable C. albicans, and was in part mediated by C. albicans matrix. However, the growth or sensitivity to amphotericin B of C. albicans is not altered in the polymicrobial biofilm. PMID- 19564371 TI - Role of the Mycobacterium tuberculosis P55 efflux pump in intrinsic drug resistance, oxidative stress responses, and growth. AB - Bacterial efflux pumps have traditionally been studied as low-level drug resistance determinants. Recent insights have suggested that efflux systems are often involved with fundamental cellular physiological processes, suggesting that drug extrusion may be a secondary function. In Mycobacterium tuberculosis, little is known about the physiological or drug resistance roles of efflux pumps. Using Mycobacterium bovis BCG as a model system, we showed that deletion of the Rv1410c gene encoding the P55 efflux pump made the strain more susceptible to a range of toxic compounds, including rifampin (rifampicin) and clofazimine, which are first and second-line antituberculosis drugs. The efflux pump inhibitors carbonyl cyanide m-chlorophenylhydrazone (CCCP) and valinomycin inhibited the P55 determined drug resistance, suggesting the active export of the compounds by use of the transmembrane proton and electrochemical gradients as sources of energy. In addition, the P55 efflux pump mutant was more susceptible to redox compounds and displayed increased intracellular redox potential, suggesting an essential role of the efflux pump in detoxification processes coupled to oxidative balance within the cell. Finally, cells that lacked the p55 gene displayed smaller colony sizes and had a growth defect in liquid culture. This, together with an increased susceptibility to the cell wall-targeting compounds bacitracin and vancomycin, suggested that P55 is needed for proper cell wall assembly and normal growth in vitro. Thus, P55 plays a fundamental role in oxidative stress responses and in vitro cell growth, in addition to contributing to intrinsic antibiotic resistance. Inhibitors of the P55 efflux pump could help to improve current treatments for tuberculosis. PMID- 19564372 TI - Vancomycin and daptomycin pharmacodynamics differ against a site-directed Staphylococcus epidermidis mutant displaying the small-colony-variant phenotype. AB - Catheter-related bloodstream infections due to slow-growing Staphylococcus epidermidis small-colony variants (SCVs) are extremely difficult to treat. Daptomycin and vancomycin pharmacodynamics were evaluated against a site-directed hemB mutant of S. epidermidis displaying the SCV phenotype and compared to that of the parental strain. The maximal killing effect decreased by 7.7-fold for vancomycin and 1.5-fold for daptomycin against the SCV mutant and were well characterized by a Hill-type mathematical model (R2 > 0.97). PMID- 19564373 TI - Infection with Anaplasma phagocytophilum induces multilineage alterations in hematopoietic progenitor cells and peripheral blood cells. AB - Infection with Anaplasma phagocytophilum, a gram-negative, lipopolysaccharide (LPS)-negative, obligate intracellular bacterium, results in multiple peripheral blood cytopenias. We hypothesized that infection with this organism would result in decreased bone marrow (BM) function and shifts in hematopoietic progenitor cells (HPCs) and lineage-committed cells in a well-established murine model of infection. HPCs and lineage-committed progenitors were enumerated in the BM and spleen during acute infection. BM cytokine production and BM CXCL12 expression were determined. Infection resulted in peripheral blood bicytopenia, marked decreases in the number of lineage-committed HPCs in the BM along with concurrent increases in the number of lineage-committed HPCs in the spleen, and a mixed, predominantly myelosuppressive BM cytokine environment. There was significant downregulation of CXCL12 in BM cells that may have been partially responsible for changes in HPC trafficking observed. Changes occurred in the absence of direct pathogen infection of BM cells. Hematopoietic lineage assessment demonstrated that there was loss of erythrocytes and B lymphocytes from the BM along with increased granulopoiesis. These changes were accompanied by splenomegaly due to lymphoid hyperplasia and increased hematopoiesis, most notably erythropoiesis. These changes largely mimic well-described inflammation and endotoxin-mediated effects on the BM and spleen; however, the numbers of peripheral blood neutrophils appear to be independently modulated as granulocytic hyperplasia does not result in neutrophilia. Our findings highlight a well-conserved series of events that we demonstrate can be instigated by an LPS-negative pathogen in the absence of an endotoxin-mediated acute proinflammatory response. PMID- 19564374 TI - Blockade of CD137 signaling counteracts polymicrobial sepsis induced by cecal ligation and puncture. AB - Sepsis, a leading cause of death worldwide, involves proinflammatory responses and inefficient bacterial clearance. Previously, we have shown that CD137 (4 1BB), a member of the tumor necrosis factor receptor superfamily, plays critical roles in eradicating infective Listeria monocytogenes, a gram-positive bacterium, and that stimulation of CD137 protects mice from sepsis-induced death. In this study, we unexpectedly found that CD137 activation aggravated polymicrobial sepsis due to mixed gram-positive and gram-negative bacterial infection induced by cecal ligation and puncture (CLP). CD137-deficient (CD137(-/-)) mice showed significantly lower mortality than CD137-sufficient (CD137(+/+)) mice in the CLP model. Administration of an agonistic anti-CD137 monoclonal antibody (MAb) to CD137(+/+) mice decreased their survival in this infection model, while administration of a blocking anti-CD137 ligand MAb (TKS-1) to such mice increased their survival. CD137(-/-) mice and TKS-1-treated CD137(+/+) mice had lower levels of chemokines/proinflammatory cytokines (monocyte chemoattractant protein 1, interleukin-6 [IL-6], tumor necrosis factor alpha, IL-12) and an anti inflammatory cytokine (IL-10), exhibited improved bacterial clearance in the peritoneum, liver, and blood, and had greater numbers of infiltrated peritoneal neutrophils and macrophages in the CLP model than control mice. Our data suggest that CD137 activation aggravates polymicrobial sepsis induced by CLP. PMID- 19564375 TI - Protection against intestinal amebiasis by a recombinant vaccine is transferable by T cells and mediated by gamma interferon. AB - We have previously shown that vaccination with purified Entamoeba histolytica Gal/GalNAc lectin or recombinant subunits can protect mice from intestinal amebiasis upon intracecal challenge. In this study, we demonstrated with adoptive transfer experiments that this lectin vaccine protection is mediated by T cells but not serum. The cell-mediated immune (CMI) response was characterized by significant gamma interferon (IFN-gamma), interleukin 12 (IL-12), IL-2, IL-10, and IL-17 production. To move toward a human vaccine, we switched to a recombinant protein and tested a range of adjuvants and routes appropriate for humans. We found that subcutaneous delivery of LecA with IDRI's adjuvant system EM014 elicited a potent Th1-type CMI profile and provided significant protection, as measured by culture negativity (79% efficacy); intranasal immunization with cholera toxin provided 56% efficacy; and alum induced a Th2-type response that protected 62 to 68% of mice. Several antibody and CMI cytokine responses were examined for correlates of protection, and prechallenge IFN-gamma(+) or IFN-gamma , IL-2-, and tumor necrosis factor alpha-triple-positive CD4 cells in blood were statistically associated with protection. To test the role of IFN-gamma in LecA mediated protection, we neutralized IFN-gamma in LecA-immunized mice and found that it abrogated the protection conferred by vaccination. These data demonstrate that CMI is sufficient for vaccine protection from intestinal amebiasis and reveal an important role for IFN-gamma, even in the setting of alum. PMID- 19564376 TI - Strain-specific duffy binding protein antibodies correlate with protection against infection with homologous compared to heterologous plasmodium vivax strains in Papua New Guinean children. AB - Individuals repeatedly infected with malaria acquire protection from infection and disease; immunity is thought to be primarily antibody-mediated and directed to blood-stage infection. Merozoite surface proteins involved in the invasion of host erythrocytes are likely targets of protective antibodies. We hypothesized that Papua New Guinean children (n = 206) who acquire high antibody levels to two Plasmodium vivax merozoite proteins, Duffy binding protein region II (PvDBPII) and the 19-kDa C-terminal region of P. vivax merozoite surface protein 1 (PvMSP1(19)), would have a delay in the time to reinfection following treatment to clear all blood-stage malaria infections. Ninety-four percent of the children were reinfected with P. vivax during biweekly follow-ups for 6 months. Since PvDBPII is polymorphic, we examined whether individuals acquired strain-specific immunity to PvDBPII. Children with high antibody levels to a prevalent PvDBPII allele (O) were associated with a delay in the time to reinfection with the same variant of P. vivax by 25% compared to parasites expressing other PvDBPII alleles (age-adjusted hazard ratio, 0.75 [95% confidence interval, 0.56 to 1.00 by Cox regression]) and 39% lower incidence density parasitemia (P = 0.01). Two other prevalent alleles (AH and P) showed a similar trend of 16% and 18% protection, respectively, against parasites with the same PvDBPII allele and reduced incidence density parasitemia. Antibodies directed to PvDBPII PNG-P and -O were both associated with a 21 to 26% reduction in the risk of P. vivax infections with higher levels of parasitemia (>150 parasites/mul), respectively. There was no association with high antibody levels to PvMSP1(19) and a delay in the time to P. vivax reinfection. Thus, anti-PvDBPII antibodies are associated with strain specific immunity to P. vivax and support the use of PvDBPII for a vaccine against P. vivax. PMID- 19564377 TI - The NanA neuraminidase of Streptococcus pneumoniae is involved in biofilm formation. AB - Streptococcus pneumoniae remains a major cause of bacteremia, pneumonia, and otitis media despite vaccines and effective antibiotics. The neuraminidase of S. pneumoniae, which catalyzes the release of terminal sialic acid residues from glycoconjugates, is involved in host colonization in animal models of infection and may provide a novel target for preventing pneumococcal infection. We demonstrate that the S. pneumoniae neuraminidase (NanA) cleaves sialic acid and show that it is involved in biofilm formation, suggesting an additional role in pathogenesis, and that it shares this property with the neuraminidase of Pseudomonas aeruginosa even though we show that the two enzymes are phylogenetically divergent. Using an in vitro model of biofilm formation incorporating human airway epithelial cells, we demonstrate that small-molecule inhibitors of NanA block biofilm formation and may provide a novel target for preventative therapy. This work highlights the role played by the neuraminidase in pathogenesis and represents an important step in drug development for prevention of colonization of the respiratory tract by this important pathogen. PMID- 19564378 TI - Bicarbonate Induces Vibrio cholerae virulence gene expression by enhancing ToxT activity. AB - Vibrio cholerae is a gram-negative bacterium that is the causative agent of cholera, a severe diarrheal illness. The two biotypes of V. cholerae O1 capable of causing cholera, classical and El Tor, require different in vitro growth conditions for induction of virulence gene expression. Growth under the inducing conditions or infection of a host initiates a complex regulatory cascade that results in production of ToxT, a regulatory protein that directly activates transcription of the genes encoding cholera toxin (CT), toxin-coregulated pilus (TCP), and other virulence genes. Previous studies have shown that sodium bicarbonate induces CT expression in the V. cholerae El Tor biotype. However, the mechanism for bicarbonate-mediated CT induction has not been defined. In this study, we demonstrate that bicarbonate stimulates virulence gene expression by enhancing ToxT activity. Both the classical and El Tor biotypes produce inactive ToxT protein when they are cultured statically in the absence of bicarbonate. Addition of bicarbonate to the culture medium does not affect ToxT production but causes a significant increase in CT and TCP expression in both biotypes. Ethoxyzolamide, a potent carbonic anhydrase inhibitor, inhibits bicarbonate mediated virulence induction, suggesting that conversion of CO(2) into bicarbonate by carbonic anhydrase plays a role in virulence induction. Thus, bicarbonate is the first positive effector for ToxT activity to be identified. Given that bicarbonate is present at high concentration in the upper small intestine where V. cholerae colonizes, bicarbonate is likely an important chemical stimulus that V. cholerae senses and that induces virulence during the natural course of infection. PMID- 19564379 TI - Macrophage-mediated responses to Candida albicans in mice expressing the human immunodeficiency virus type 1 transgene. AB - The critical impairments of innate and adaptive immunity that cause susceptibility to mucosal candidiasis in human immunodeficiency virus (HIV) infection have not been fully determined. We therefore conducted an analysis of macrophage-mediated responses to Candida albicans in transgenic (Tg) mice expressing Nef, Env, and Rev of HIV type 1 (HIV-1) in CD4(+) T cells, dendritic cells, and macrophages and developing an AIDS-like disease (CD4C/HIV(MutA) Tg mice). Macrophages were successfully recruited to the oral and gastric mucosae of these Tg mice in response to chronic carriage of C. albicans and displayed polarization toward an alternatively activated phenotype. Functionally, peritoneal macrophages from uninfected Tg mice exhibited increased phagocytosis of C. albicans and enhanced production of interleukin 6 and monocyte chemoattractant protein 1, demonstrating that the HIV-1 transgene independently activates selected macrophage functions. Production of H(2)O(2) by macrophages from Tg mice primed with gamma interferon and treated with phorbol 12-myristate 13-acetate or C. albicans was moderately reduced, but expression of the HIV-1 transgene did not alter production of nitric oxide or reduce killing of C. albicans. A knockout of the inducible nitric oxide synthase (NOS2) gene in these Tg mice did not augment oral or gastrointestinal burdens during chronic carriage of C. albicans or cause systemic dissemination, likely due to a redundancy provided by partially preserved production of H(2)O(2) and oxygen-independent candidacidal mechanisms. Thus, the macrophage response to C. albicans is largely preserved in these Tg mice, and no functional macrophage defect appears to primarily determine the susceptibility to mucosal candidiasis. PMID- 19564380 TI - Transmembrane tumor necrosis factor alpha is required for enteropathy and is sufficient to promote parasite expulsion in gastrointestinal helminth infection. AB - To study the specific role of transmembrane tumor necrosis factor (tmTNF) in protective and pathological responses against the gastrointestinal helminth Trichinella spiralis, we compared the immune responses of TNF-alpha/lymphotoxin alpha (LTalpha)(-/-) mice expressing noncleavable transgenic tmTNF to those of TNF-alpha/LTalpha(-/-) and wild-type mice. The susceptibility of TNF alpha/LTalpha(-/-) mice to T. spiralis infection was associated with impaired induction of a protective Th2 response and the lack of mucosal mastocytosis. Although tmTNF-expressing transgenic (tmTNF-tg) mice also had a reduced Th2 response, the mast cell response was greater than that observed in TNF alpha/LTalpha(-/-) mice and was sufficient to induce the expulsion of the parasite. T. spiralis infection of tmTNF-tg mice resulted in significant intestinal pathology characterized by villus atrophy and crypt hyperplasia comparable to that induced following the infection of wild-type mice, while pathology in TNF-alpha/LTalpha(-/-) mice was significantly reduced. Our data thus indicate a role for tmTNF in host defense against gastrointestinal helminths and in the accompanying enteropathy. Furthermore, they also demonstrate that TNF alpha is required for the induction of Th2 immune responses related to infection with gastrointestinal helminth parasites. PMID- 19564381 TI - LuxS promotes biofilm maturation and persistence of nontypeable haemophilus influenzae in vivo via modulation of lipooligosaccharides on the bacterial surface. AB - Nontypeable Haemophilus influenzae (NTHI) is an extremely common airway commensal which can cause opportunistic infections that are usually localized to airway mucosal surfaces. During many of these infections, NTHI forms biofilm communities that promote persistence in vivo. For many bacterial species, density-dependent quorum-signaling networks can affect biofilm formation and/or maturation. Mutation of luxS, a determinant of the autoinducer 2 (AI-2) quorum signal pathway, increases NTHI virulence in the chinchilla model for otitis media infections. For example, bacterial counts in middle-ear fluids and the severity of the host inflammatory response were increased in luxS mutants compared with parental strains. As these phenotypes are consistent with those that we have observed for biofilm-defective NTHI mutants, we hypothesized that luxS may affect NTHI biofilms. A luxS mutant was generated using the well-characterized NTHI 86 028NP strain and tested to determine the effects of the mutation on biofilm phenotypes in vitro and bacterial persistence and disease severity during experimental otitis media. Quantitation of the biofilm structure by confocal microscopy and COMSTAT analysis revealed significantly reduced biomass for NTHI 86-028NP luxS biofilms, which was restored by a soluble mediator in NTHI 86-028NP supernatants. Analysis of lipooligosaccharide moieties using an enzyme-linked immunosorbent assay and immunoblotting showed decreased levels of biofilm associated glycoforms in the NTHI 86-028NP luxS strain. Infection studies showed that NTHI 86-028NP luxS had a significant persistence defect in vivo during chronic otitis media infection. Based on these data, we concluded that a luxS dependent soluble mediator modulates the composition of the NTHI lipooligosaccharides, resulting in effects on biofilm maturation and bacterial persistence in vivo. PMID- 19564382 TI - Antibiotic treatment of clostridium difficile carrier mice triggers a supershedder state, spore-mediated transmission, and severe disease in immunocompromised hosts. AB - Clostridium difficile persists in hospitals by exploiting an infection cycle that is dependent on humans shedding highly resistant and infectious spores. Here we show that human virulent C. difficile can asymptomatically colonize the intestines of immunocompetent mice, establishing a carrier state that persists for many months. C. difficile carrier mice consistently shed low levels of spores but, surprisingly, do not transmit infection to cohabiting mice. However, antibiotic treatment of carriers triggers a highly contagious supershedder state, characterized by a dramatic reduction in the intestinal microbiota species diversity, C. difficile overgrowth, and excretion of high levels of spores. Stopping antibiotic treatment normally leads to recovery of the intestinal microbiota species diversity and suppresses C. difficile levels, although some mice persist in the supershedding state for extended periods. Spore-mediated transmission to immunocompetent mice treated with antibiotics results in self limiting mucosal inflammation of the large intestine. In contrast, transmission to mice whose innate immune responses are compromised (Myd88(-/-)) leads to a severe intestinal disease that is often fatal. Thus, mice can be used to investigate distinct stages of the C. difficile infection cycle and can serve as a valuable surrogate for studying the spore-mediated transmission and interactions between C. difficile and the host and its microbiota, and the results obtained should guide infection control measures. PMID- 19564383 TI - Sialic acid catabolism confers a competitive advantage to pathogenic vibrio cholerae in the mouse intestine. AB - Sialic acids comprise a family of nine-carbon ketosugars that are ubiquitous on mammalian mucous membranes. However, sialic acids have a limited distribution among Bacteria and are confined mainly to pathogenic and commensal species. Vibrio pathogenicity island 2 (VPI-2), a 57-kb region found exclusively among pathogenic strains of Vibrio cholerae, contains a cluster of genes (nan-nag) putatively involved in the scavenging (nanH), transport (dctPQM), and catabolism (nanA, nanE, nanK, and nagA) of sialic acid. The capacity to utilize sialic acid as a carbon and energy source might confer an advantage to V. cholerae in the mucus-rich environment of the gut, where sialic acid availability is extensive. In this study, we show that V. cholerae can utilize sialic acid as a sole carbon source. We demonstrate that the genes involved in the utilization of sialic acid are located within the nan-nag region of VPI-2 by complementation of Escherichia coli mutants and gene knockouts in V. cholerae N16961. We show that nanH, dctP, nanA, and nanK are highly expressed in V. cholerae grown on sialic acid. By using the infant mouse model of infection, we show that V. cholerae DeltananA strain SAM1776 is defective in early intestinal colonization stages. In addition, SAM1776 shows a decrease in the competitive index in colonization-competition assays comparing the mutant strain with both O1 El Tor and classical strains. Our data indicate an important relationship between the catabolism of sialic acid and bacterial pathogenesis, stressing the relevance of the utilization of the resources found in the host's environment. PMID- 19564385 TI - Transcription factor PrtT controls expression of multiple secreted proteases in the human pathogenic mold Aspergillus fumigatus. AB - The role of secreted proteases in the virulence of the pathogenic fungus Aspergillus fumigatus remains controversial. Recently, the Aspergillus niger transcription factor PrtT was shown to control the expression of multiple secreted proteases. In this work, the gene which encodes the PrtT homolog in A. fumigatus was cloned and its function analyzed using a deletion mutant strain. Deletion of A. fumigatus prtT resulted in the loss of secreted protease activity. The expression of six secreted proteases (ALP, MEP, Dpp4, CpdS, AFUA_2G17330, and AFUA_7G06220) was markedly reduced. Culture filtrates from the prtT deletion strain exhibited reduced killing of lung epithelial cells and lysis of erythrocytes. However, the prtT deletion strain did not exhibit altered virulence in lung-infected mice. These results suggest that PrtT is not a significant virulence factor in A. fumigatus. PMID- 19564384 TI - Staphylococcal alpha-toxin is not sufficient to mediate escape from phagolysosomes in upper-airway epithelial cells. AB - Intracellular Staphylococcus aureus has been implicated in the establishment of chronic infections. It is therefore imperative to understand by what means S. aureus is able to survive within cells. Here we use two expression systems with a fluorescent readout to assay alpha-toxin expression and function within phagolysosomes of infected upper-airway epithelial cells: avirulent Staphylococcus carnosus TM300 and phenotypically alpha-toxin-negative S. aureus laboratory strains. Data from CFU recovery assays suggest that the presence of alpha-toxin is not beneficial for the intracellular survival of recombinant Staphylococcus strains. This finding was corroborated by immunofluorescence studies: whereas S. carnosus and S. aureus are able to deliver S. aureus alpha toxin to lumina of host cell phagolysosomes, the membrane integrity of these organelles was not affected. Alpha-toxin-expressing strains were detected exclusively within lysosome-associated membrane protein 1 (LAMP1)-yellow fluorescent protein (YFP)-positive vesicles. Measurements of intraphagosomal pH illustrated that all infected phagolysosomes acidified regardless of alpha-toxin expression. In contrast, S. aureus expressing Listeria monocytogenes listeriolysin O leads to the breakdown of the phagolysosomal membrane, as indicated by staphylococci that are not associated with LAMP1-YFP-decorated vesicles and that do not reside within an acidic cellular environment. Thus, our results suggest that staphylococcal alpha-toxin is not sufficient to mediate phagolysosomal escape in upper-airway epithelial cells. PMID- 19564386 TI - CD4+ CD25+ Foxp3- T-regulatory cells produce both gamma interferon and interleukin-10 during acute severe murine spotted fever rickettsiosis. AB - Spotted fever group rickettsiae cause life-threatening human infections worldwide. Until now, the immune regulatory mechanisms involved in fatal rickettsial infection have been unknown. C3H/HeN mice infected with 3 x 10(5) PFU of Rickettsia conorii developed an acute progressive disease, and all mice succumbed to this infection. A sublethal infection induced protective immunity, and mice survived. Compared to splenic T cells from sublethally infected mice, splenic T cells from lethally infected mice produced significantly lower levels of interleukin-2 (IL-2) and gamma interferon (IFN-gamma) and a higher level of IL 10, but not of IL-4 or transforming growth factor beta, and there was markedly suppressed CD4(+) T-cell proliferation in response to antigen-specific stimulation with R. conorii. Furthermore, lethal infection induced significant expansion of CD4(+) CD25(+) Foxp3(-) T cells in infected organs compared to the levels in naive and sublethally infected mice. In a lethal infection, splenic CD4(+) CD25(+) Foxp3(-) T cells, which were CTLA-4(high) T-bet(+) and secreted both IFN-gamma and IL-10, suppressed the proliferation of and IL-2 production by splenic CD4(+) CD25(-) Foxp3(-) T cells in vitro. Interestingly, depletion of CD25(+) T cells in vivo did not change the disease progression, but it increased the bacterial load in the lung and liver, significantly reduced the number of IFN gamma-producing Th1 cells in the spleen, and increased the serum levels of IFN gamma. These results suggested that CD4(+) CD25(+) T cells generated in acute murine spotted fever rickettsiosis are Th1-cell-related adaptive T-regulatory cells, which substantially contribute to suppressing the systemic immune response, possibly by a mechanism involving IL-10 and/or cytotoxic T-lymphocyte antigen 4. PMID- 19564387 TI - Aggregatibacter actinomycetemcomitans builds mutualistic biofilm communities with Fusobacterium nucleatum and Veillonella species in saliva. AB - Human oral bacterial pathogens grow in attached multispecies biofilm communities. Unattached cells are quickly removed by swallowing. Therefore, surface attachment is essential for growth, and we investigated multispecies community interactions resulting in mutualistic growth on saliva as the sole nutritional source. We used two model systems, saliva-coated transferable solid-phase polystyrene pegs (peg biofilms) and flow cells with saliva-coated glass surfaces. Fluorescent antibody staining and image analysis were used to quantify the biomass in flow cells, and quantitative real-time PCR with species-specific primers was used to quantify the biomass in peg biofilms. Veillonella sp. strain PK1910, Aggregatibacter actinomycetemcomitans JP2, and Fusobacterium nucleatum ATCC 10953 were unable to grow as single species in flow cells. Only A. actinomycetemcomitans grew after 36 h when peg biofilms remained submerged in saliva from the time of inoculation. Mixed-species coaggregates were used for two- and three-species inoculation. The biomass in two-species biofilms increased in both systems when Veillonella sp. strain PK1910 was present as one of the partners. Enhanced growth of all strains was observed in three-species biofilms in flow cells. Interestingly, in flow cells F. nucleatum and A. actinomycetemcomitans exhibited mutualism, and, although F. nucleatum was unable to grow with either of the other species in the peg system, F. nucleatum stimulated the growth of Veillonella sp. and together these two organisms increased the total biomass of A. actinomycetemcomitans in three-species peg biofilms. We propose that mutualistic two-species and multispecies oral biofilm communities form in vivo and that mutualism between commensal veillonellae and late colonizing pathogens, such as aggregatibacteria, contributes to the development of periodontal disease. PMID- 19564388 TI - Role of dendritic cells and alveolar macrophages in regulating early host defense against pulmonary infection with Cryptococcus neoformans. AB - Successful pulmonary clearance of the encapsulated yeast Cryptococcus neoformans requires a T1 adaptive immune response. This response takes up to 3 weeks to fully develop. The role of the initial, innate immune response against the organism is uncertain. In this study, an established model of diphtheria toxin mediated depletion of resident pulmonary dendritic cells (DC) and alveolar macrophages (AM) was used to assess the contribution of these cells to the initial host response against cryptococcal infection. The results demonstrate that depletion of DC and AM one day prior to infection results in rapid clinical deterioration and death of mice within 6 days postinfection; this effect was not observed in infected groups of control mice not depleted of DC and AM. Depletion did not alter the microbial burden or total leukocyte recruitment in the lung. Mortality (in mice depleted of DC and AM) was associated with increased neutrophil and B-cell accumulation accompanied by histopathologic evidence of suppurative neutrophilic bronchopneumonia, cyst formation, and alveolar damage. Collectively, these data define an important role for DC and AM in regulating the initial innate immune response following pulmonary infection with C. neoformans. These findings provide important insight into the cellular mechanisms which coordinate early host defense against an invasive fungal pathogen in the lung. PMID- 19564389 TI - Analysis of the genome of the Escherichia coli O157:H7 2006 spinach-associated outbreak isolate indicates candidate genes that may enhance virulence. AB - In addition to causing diarrhea, Escherichia coli O157:H7 infection can lead to hemolytic-uremic syndrome (HUS), a severe disease characterized by hemolysis and renal failure. Differences in HUS frequency among E. coli O157:H7 outbreaks have been noted, but our understanding of bacterial factors that promote HUS is incomplete. In 2006, in an outbreak of E. coli O157:H7 caused by consumption of contaminated spinach, there was a notably high frequency of HUS. We sequenced the genome of the strain responsible (TW14359) with the goal of identifying candidate genetic factors that contribute to an enhanced ability to cause HUS. The TW14359 genome contains 70 kb of DNA segments not present in either of the two reference O157:H7 genomes. We identified seven putative virulence determinants, including two putative type III secretion system effector proteins, candidate genes that could result in increased pathogenicity or, alternatively, adaptation to plants, and an intact anaerobic nitric oxide reductase gene, norV. We surveyed 100 O157:H7 isolates for the presence of these putative virulence determinants. A norV deletion was found in over one-half of the strains surveyed and correlated strikingly with the absence of stx(1). The other putative virulence factors were found in 8 to 35% of the O157:H7 isolates surveyed, and their presence also correlated with the presence of norV and the absence of stx(1), indicating that the presence of norV may serve as a marker of a greater propensity for HUS, similar to the correlation between the absence of stx(1) and a propensity for HUS. PMID- 19564390 TI - A regulator of Aspergillus fumigatus extracellular proteolytic activity is dispensable for virulence. AB - Virulence of the fungal pathogen Aspergillus fumigatus is in part based on the saprophytic lifestyle that this mold has evolved. A crucial function for saprophytism resides in secreted proteases that allow assimilation of proteinaceous substrates. The impact of extracellular proteolytic activities on the pathogenesis of aspergillosis, however, remains controversial. In order to address this issue, characterization of a conserved regulatory factor, PrtT, that acts on expression of secreted proteases was pursued. Expression of PrtT appears to be regulated posttranscriptionally, and the existence of an mRNA leader sequence implies translational control via eIF2alpha kinase signaling. Phenotypic classification of a prtTDelta deletion mutant revealed that expression of several major extracellular proteases is PrtT dependent, resulting in the inability to utilize protein as a nutritional source. Certain genes encoding secreted proteases are not regulated by PrtT. Most strikingly, the deletant strain is not attenuated in virulence when tested in a leukopenic mouse model, which makes a strong case for reconsidering any impact of secreted proteases in pulmonary aspergillosis. PMID- 19564391 TI - Roles of RseB, sigmaE, and DegP in virulence and phase variation of colony morphotype of Vibrio vulnificus. AB - Vibrio vulnificus is an estuarine bacterium capable of causing serious and often fatal wound infections and primary septicemia. We used alkaline phosphatase insertion mutagenesis to identify genes necessary for the virulence of this pathogen. One mutant had an in-frame fusion of 'phoA to the gene encoding RseB, a periplasmic negative regulator of the alternative sigma factor sigma(E). sigma(E) controls an extensive regulon involved in responding to cell envelope stresses. Colonies of the rseB mutant were less opaque than wild-type colonies and underwent phase variation between translucent and opaque morphologies. rseB mutants were attenuated for virulence in subcutaneously inoculated iron-dextran treated mice. To obtain insight into the role of rseB and the extracytoplasmic stress response in V. vulnificus, mutants with defined mutations in rseB and two important members of the extracytoplasmic stress regulon, rpoE and degP, were constructed for analysis of virulence, colony morphology, and stress-associated phenotypes. Deletion of rseB caused reversible phase variation in the colony morphotype that was associated with extracellular polysaccharides. Translucent and transparent morphotype strains were attenuated for virulence. rpoE and degP deletion mutants were sensitive to membrane-perturbing agents and heat but were not significantly attenuated for V. vulnificus virulence in mice. These results reveal complex relationships between regulation of the extracytoplasmic stress response, exopolysaccharides, and the virulence of V. vulnificus. PMID- 19564392 TI - Toxoplasma gondii cyclophilin 18-mediated production of nitric oxide induces Bradyzoite conversion in a CCR5-dependent manner. AB - Toxoplasma gondii modulates pro- and anti-inflammatory responses to regulate parasite multiplication and host survival. Pressure from the immune response causes the conversion of tachyzoites into slowly dividing bradyzoites. The regulatory mechanisms involved in this switch are poorly understood. The aim of this study was to investigate the immunomodulatory role of T. gondii cyclophilin 18 (TgCyp18) in macrophages and the consequences of the cellular responses on the conversion machinery. Recombinant TgCyp18 induced the production of nitric oxide (NO), interleukin-12 (IL-12), and tumor necrosis factor alpha through its binding with cysteine-cysteine chemokine receptor 5 (CCR5) and the production of gamma interferon and IL-6 in a CCR5-independent manner. Interestingly, the treatment of macrophages with TgCyp18 resulted in the inhibition of parasite growth and an enhancement of the conversion into bradyzoites via NO in a CCR5-dependent manner. In conclusion, T. gondii possesses sophisticated mechanisms to manipulate host cell responses in a TgCyp18-mediated process. PMID- 19564395 TI - A simple two-stage model predicts response time distributions. AB - The neural mechanisms underlying reaction times have previously been modelled in two distinct ways. When stimuli are hard to detect, response time tends to follow a random-walk model that integrates noisy sensory signals. But studies investigating the influence of higher-level factors such as prior probability and response urgency typically use highly detectable targets, and response times then usually correspond to a linear rise-to-threshold mechanism. Here we show that a model incorporating both types of element in series - a detector integrating noisy afferent signals, followed by a linear rise-to-threshold performing decision - successfully predicts not only mean response times but, much more stringently, the observed distribution of these times and the rate of decision errors over a wide range of stimulus detectability. By reconciling what previously may have seemed to be conflicting theories, we are now closer to having a complete description of reaction time and the decision processes that underlie it. PMID- 19564394 TI - Homocysteine transport by systems L, A and y+L across the microvillous plasma membrane of human placenta. AB - Elevated maternal plasma levels of homocysteine (Hcy) are associated with pregnancy complications and adverse neonatal outcomes, suggesting placental transport of Hcy may impact on fetal development. However, such transport mechanisms have not been defined. In this study we characterise Hcy transport mechanisms across the microvillous plasma membrane (MVM) of the syncytiotrophoblast, the transporting epithelium of human placenta. Three candidate transport systems, systems L, A and y(+)L, were examined utilising competitive inhibition to investigate the effects of Hcy on the uptake of well characterised radiolabelled substrates for each system into isolated MVM vesicles, and that of model substrates on 10 microm [(35)S]l-Hcy uptake. System L activity was inhibited by both l-Hcy and dl-Hcy, comparable to model substrates including 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid (BCH). System L constituted the major transport mechanism, with significant BCH inhibition (69%) of [(35)S]l-Hcy uptake. System A activity was also inhibited by l-Hcy and dl-Hcy with a smaller contribution (21%) to [(35)S]l-Hcy uptake. Inhibition by l-Hcy and dl-Hcy of system y(+)L activity was Na(+) sensitive with a significant inhibition constant (K(i)) shift observed following K(+) replacement; l-arginine reduced [(35)S]l-Hcy uptake by 19%. Kinetic modelling of [(35)S]l-Hcy uptake resolved two, Na(+)-independent, transport components (K(m) 72 microm and 9.7 mm). This study provides evidence for the involvement of systems L, A and y(+)L in placental Hcy transport. Such transport, by competing with endogenous amino acids for transporter activity, could have major implications for syncytiotrophoblast metabolism and function as well as fetal development. PMID- 19564396 TI - Caffeine inhibition of ionotropic glycine receptors. AB - We found that caffeine is a structural analogue of strychnine and a competitive antagonist at ionotropic glycine receptors (GlyRs). Docking simulations indicate that caffeine and strychnine may bind to similar sites at the GlyR. The R131A GlyR mutation, which reduces strychnine antagonism without suppressing activation by glycine, also reduces caffeine antagonism. GlyR subtypes have differing caffeine sensitivity. Tested against the EC(50) of each GlyR subtype, the order of caffeine potency (IC(50)) is: alpha2beta (248 +/- 32 microm) alpha3beta (255 +/- 16 microm) > alpha4beta (517 +/- 50 microm) > alpha1beta(837 +/- 132 microm). However, because the alpha3beta GlyR is more than 3-fold less sensitive to glycine than any of the other GlyR subtypes, this receptor is most effectively blocked by caffeine. The glycine dose-response curves and the effects of caffeine indicate that amphibian retinal ganglion cells do not express a plethora of GlyR subtypes and are dominated by the alpha1beta GlyR. Comparing the effects of caffeine on glycinergic spontaneous and evoked IPSCs indicates that evoked release elevates the glycine concentration at some synapses whereas summation elicits evoked IPSCs at other synapses. Caffeine serves to identify the pharmacophore of strychnine and produces near-complete inhibition of glycine receptors at concentrations commonly employed to stimulate ryanodine receptors. PMID- 19564393 TI - Pathophysiological actions of neuropathy-related anti-ganglioside antibodies at the neuromuscular junction. AB - The outer leaflet of neuronal membranes is highly enriched in gangliosides. Therefore, specific neuronal roles have been attributed to this family of sialylated glycosphingolipids, e.g. in modulation of ion channels and transporters, neuronal interaction and recognition, temperature adaptation, Ca(2+) homeostasis, axonal growth, (para)node of Ranvier stability and synaptic transmission. Recent developmental, ageing and injury studies on transgenic mice lacking subsets of gangliosides indicate that gangliosides are involved in maintenance rather than development of the nervous system and that ganglioside family members are able to act in a mutually compensatory manner. Besides having physiological functions, gangliosides are the likely antigenic targets of autoantibodies present in Guillain-Barre syndrome (GBS), a group of neuropathies with clinical symptoms of motor- and/or sensory peripheral nerve dysfunction. Antibody binding to peripheral nerves is thought to either interfere with ganglioside function or activate complement, causing axonal damage and thereby disturbed action potential conduction. The presynaptic motor nerve terminal at the neuromuscular junction (NMJ) may be a prominent target because it is highly enriched in gangliosides and lies outside the blood-nerve barrier, allowing antibody access. The ensuing neuromuscular synaptopathy might contribute to the muscle weakness in GBS patients. Several groups, including our own, have studied the effects of anti-ganglioside antibodies in ex vivo and in vivo experimental settings at mouse NMJs. Here, after providing a background overview on ganglioside synthesis, localization and physiology, we will review those studies, which clearly show that anti-ganglioside antibodies are capable of binding to NMJs and thereby can exert a variety of pathophysiological effects. Furthermore, we will discuss the human clinical electrophysiological and histological evidence produced so far of the existence of a neuromuscular synaptopathy contributing to muscle weakness in GBS patients. PMID- 19564397 TI - Upregulation of inward rectifier K+ (Kir2) channels in dentate gyrus granule cells in temporal lobe epilepsy. AB - In humans, temporal lobe epilepsy (TLE) is often associated with Ammon's horn sclerosis (AHS) characterized by hippocampal cell death, gliosis and granule cell dispersion (GCD) in the dentate gyrus. Granule cells surviving TLE have been proposed to be hyperexcitable and to play an important role in seizure generation. However, it is unclear whether this applies to conditions of AHS. We studied granule cells using the intrahippocampal kainate injection mouse model of TLE, brain slice patch-clamp recordings, morphological reconstructions and immunocytochemistry. With progressing AHS and GCD, 'epileptic' granule cells of the injected hippocampus displayed a decreased input resistance, a decreased membrane time constant and an increased rheobase. The resting leak conductance was doubled in epileptic granule cells and roughly 70-80% of this difference were sensitive to K(+) replacement. Of the increased K(+) leak, about 50% were sensitive to 1 mm Ba(2+). Approximately 20-30% of the pathological leak was mediated by a bicuculline-sensitive GABA(A) conductance. Epileptic granule cells had strongly enlarged inwardly rectifying currents with a low micromolar Ba(2+) IC(50), reminiscent of classic inward rectifier K(+) channels (Irk/Kir2). Indeed, protein expression of Kir2 subunits (Kir2.1, Kir2.2, Kir2.3, Kir2.4) was upregulated in epileptic granule cells. Immunolabelling for two-pore weak inward rectifier K(+) channels (Twik1/K2P1.1, Twik2/K2P6.1) was also increased. We conclude that the excitability of granule cells in the sclerotic focus of TLE is reduced due to an increased resting conductance mainly due to upregulated K(+) channel expression. These results point to a local adaptive mechanism that could counterbalance hyperexcitability in epilepsy. PMID- 19564399 TI - Antonio Giraldez: at the tip of the microRNA iceberg. PMID- 19564400 TI - Accuracy and precision in quantitative fluorescence microscopy. AB - The light microscope has long been used to document the localization of fluorescent molecules in cell biology research. With advances in digital cameras and the discovery and development of genetically encoded fluorophores, there has been a huge increase in the use of fluorescence microscopy to quantify spatial and temporal measurements of fluorescent molecules in biological specimens. Whether simply comparing the relative intensities of two fluorescent specimens, or using advanced techniques like Forster resonance energy transfer (FRET) or fluorescence recovery after photobleaching (FRAP), quantitation of fluorescence requires a thorough understanding of the limitations of and proper use of the different components of the imaging system. Here, I focus on the parameters of digital image acquisition that affect the accuracy and precision of quantitative fluorescence microscopy measurements. PMID- 19564401 TI - Motor-dependent microtubule disassembly driven by tubulin tyrosination. AB - In cells, stable microtubules (MTs) are covalently modified by a carboxypeptidase, which removes the C-terminal Tyr residue of alpha-tubulin. The significance of this selective detyrosination of MTs is not understood. In this study, we report that tubulin detyrosination in fibroblasts inhibits MT disassembly. This inhibition is relieved by overexpression of the depolymerizing motor mitotic centromere-associated kinesin (MCAK). Conversely, suppression of MCAK expression prevents disassembly of normal tyrosinated MTs in fibroblasts. Detyrosination of MTs suppresses the activity of MCAK in vitro, apparently as the result of a decreased affinity of the adenosine diphosphate (ADP)-inorganic phosphate- and ADP-bound forms of MCAK for the MT lattice. Detyrosination also impairs MT disassembly in neurons and inhibits the activity of the neuronal depolymerizing motor KIF2A in vitro. These results indicate that MT depolymerizing motors are directly inhibited by the detyrosination of tubulin, resulting in the stabilization of cellular MTs. Detyrosination of transiently stabilized MTs may give rise to persistent subpopulations of disassembly resistant polymers to sustain subcellular cytoskeletal differentiation. PMID- 19564402 TI - Distinct cytoplasmic maturation steps of 40S ribosomal subunit precursors require hRio2. AB - During their biogenesis, 40S ribosomal subunit precursors are exported from the nucleus to the cytoplasm, where final maturation occurs. In this study, we show that the protein kinase human Rio2 (hRio2) is part of a late 40S preribosomal particle in human cells. Using a novel 40S biogenesis and export assay, we analyzed the contribution of hRio2 to late 40S maturation. Although hRio2 is not absolutely required for pre-40S export, deletion of its binding site for the export receptor CRM1 decelerated the kinetics of this process. Moreover, in the absence of hRio2, final cytoplasmic 40S maturation is blocked because the recycling of several trans-acting factors and cytoplasmic 18S-E precursor ribosomal RNA (rRNA [pre-rRNA]) processing are defective. Intriguingly, the physical presence of hRio2 but not its kinase activity is necessary for the release of hEnp1 from cytoplasmic 40S precursors. In contrast, hRio2 kinase activity is essential for the recycling of hDim2, hLtv1, and hNob1 as well as for 18S-E pre-rRNA processing. Thus, hRio2 is involved in late 40S maturation at several distinct steps. PMID- 19564403 TI - Biogenesis of cytochrome b6 in photosynthetic membranes. AB - In chloroplasts, binding of a c'-heme to cytochrome b(6) on the stromal side of the thylakoid membranes requires a specific mechanism distinct from the one at work for c-heme binding to cytochromes f and c(6) on the lumenal side of membranes. Here, we show that the major protein components of this pathway, the CCBs, are bona fide transmembrane proteins. We demonstrate their association in a series of hetero-oligomeric complexes, some of which interact transiently with cytochrome b(6) in the process of heme delivery to the apoprotein. In addition, we provide preliminary evidence for functional assembly of cytochrome b(6)f complexes even in the absence of c'-heme binding to cytochrome b(6). Finally, we present a sequential model for apo- to holo-cytochrome b(6) maturation integrated within the assembly pathway of b(6)f complexes in the thylakoid membranes. PMID- 19564404 TI - Cholesterol sensor ORP1L contacts the ER protein VAP to control Rab7-RILP-p150 Glued and late endosome positioning. AB - Late endosomes (LEs) have characteristic intracellular distributions determined by their interactions with various motor proteins. Motor proteins associated to the dynactin subunit p150(Glued) bind to LEs via the Rab7 effector Rab7 interacting lysosomal protein (RILP) in association with the oxysterol-binding protein ORP1L. We found that cholesterol levels in LEs are sensed by ORP1L and are lower in peripheral vesicles. Under low cholesterol conditions, ORP1L conformation induces the formation of endoplasmic reticulum (ER)-LE membrane contact sites. At these sites, the ER protein VAP (VAMP [vesicle-associated membrane protein]-associated ER protein) can interact in trans with the Rab7-RILP complex to remove p150(Glued) and associated motors. LEs then move to the microtubule plus end. Under high cholesterol conditions, as in Niemann-Pick type C disease, this process is prevented, and LEs accumulate at the microtubule minus end as the result of dynein motor activity. These data explain how the ER and cholesterol control the association of LEs with motor proteins and their positioning in cells. PMID- 19564405 TI - A genome-wide screen for genes affecting eisosomes reveals Nce102 function in sphingolipid signaling. AB - The protein and lipid composition of eukaryotic plasma membranes is highly dynamic and regulated according to need. The sphingolipid-responsive Pkh kinases are candidates for mediating parts of this regulation, as they affect a diverse set of plasma membrane functions, such as cortical actin patch organization, efficient endocytosis, and eisosome assembly. Eisosomes are large protein complexes underlying the plasma membrane and help to sort a group of membrane proteins into distinct domains. In this study, we identify Nce102 in a genome wide screen for genes involved in eisosome organization and Pkh kinase signaling. Nce102 accumulates in membrane domains at eisosomes where Pkh kinases also localize. The relative abundance of Nce102 in these domains compared with the rest of the plasma membrane is dynamically regulated by sphingolipids. Furthermore, Nce102 inhibits Pkh kinase signaling and is required for plasma membrane organization. Therefore, Nce102 might act as a sensor of sphingolipids that regulates plasma membrane function. PMID- 19564406 TI - Demonstration of catch bonds between an integrin and its ligand. AB - Binding of integrins to ligands provides anchorage and signals for the cell, making them prime candidates for mechanosensing molecules. How force regulates integrin-ligand dissociation is unclear. We used atomic force microscopy to measure the force-dependent lifetimes of single bonds between a fibronectin fragment and an integrin alpha(5)beta(1)-Fc fusion protein or membrane alpha(5)beta(1). Force prolonged bond lifetimes in the 10-30-pN range, a counterintuitive behavior called catch bonds. Changing cations from Ca(2+)/Mg(2+) to Mg(2+)/EGTA and to Mn(2+) caused longer lifetime in the same 10-30-pN catch bond region. A truncated alpha(5)beta(1) construct containing the headpiece but not the legs formed longer-lived catch bonds that were not affected by cation changes at forces <30 pN. Binding of monoclonal antibodies that induce the active conformation of the integrin headpiece shifted catch bonds to a lower force range. Thus, catch bond formation appears to involve force-assisted activation of the headpiece but not integrin extension. PMID- 19564407 TI - The enterocyte microvillus is a vesicle-generating organelle. AB - For decades, enterocyte brush border microvilli have been viewed as passive cytoskeletal scaffolds that serve to increase apical membrane surface area. However, recent studies revealed that in the in vitro context of isolated brush borders, myosin-1a (myo1a) powers the sliding of microvillar membrane along core actin bundles. This activity also leads to the shedding of small vesicles from microvillar tips, suggesting that microvilli may function as vesicle-generating organelles in vivo. In this study, we present data in support of this hypothesis, showing that enterocyte microvilli release unilamellar vesicles into the intestinal lumen; these vesicles retain the right side out orientation of microvillar membrane, contain catalytically active brush border enzymes, and are specifically enriched in intestinal alkaline phosphatase. Moreover, myo1a knockout mice demonstrate striking perturbations in vesicle production, clearly implicating this motor in the in vivo regulation of this novel activity. In combination, these data show that microvilli function as vesicle-generating organelles, which enable enterocytes to deploy catalytic activities into the intestinal lumen. PMID- 19564408 TI - C/EBPalpha and the corepressors CtBP1 and CtBP2 regulate repression of select visceral white adipose genes during induction of the brown phenotype in white adipocytes by peroxisome proliferator-activated receptor gamma agonists. AB - White adipose tissue (WAT) stores energy in the form of triglycerides, whereas brown tissue (BAT) expends energy, primarily by oxidizing lipids. WAT also secretes many cytokines and acute-phase proteins that contribute to insulin resistance in obese subjects. In this study, we have investigated the mechanisms by which activation of peroxisome proliferator-activated receptor gamma (PPARgamma) with synthetic agonists induces a brown phenotype in white adipocytes in vivo and in vitro. We demonstrate that this phenotypic conversion is characterized by repression of a set of white fat genes ("visceral white"), including the resistin, angiotensinogen, and chemerin genes, in addition to induction of brown-specific genes, such as Ucp-1. Importantly, the level of expression of the "visceral white" genes is high in mesenteric and gonadal WAT depots but low in the subcutaneous WAT depot and in BAT. Mutation of critical amino acids within helix 7 of the ligand-binding domain of PPARgamma prevents inhibition of visceral white gene expression by the synthetic agonists and therefore shows a direct role for PPARgamma in the repression process. Inhibition of the white adipocyte genes also depends on the expression of C/EBPalpha and the corepressors, carboxy-terminal binding proteins 1 and 2 (CtBP1/2). The data further show that repression of resistin and angiotensinogen expression involves recruitment of CtBP1/2, directed by C/EBPalpha, to the minimal promoter of the corresponding genes in response to the PPARgamma ligand. Developing strategies to enhance the brown phenotype in white adipocytes while reducing secretion of stress-related cytokines from visceral WAT is a means to combat obesity associated disorders. PMID- 19564409 TI - Id1 attenuates Notch signaling and impairs T-cell commitment by elevating Deltex1 expression. AB - Complete inhibition of E protein transcription factors by Id1 blocks the developmental transition of CD4/CD8 double-negative 1 (DN1; CD44(+) CD25(-)) thymocytes to the DN2 (CD44(+) CD25(+)) stage. To understand the underlying mechanisms, we observed that mRNA levels of Deltex1, as well as Deltex4, were dramatically elevated in Id1-expressing thymocytes, which could result in developmental arrest by attenuating Notch function. In support of this hypothesis, we found that Deltex1 ablation enabled Id1-expressing progenitors to differentiate to the DN3 (CD44(-) CD25(+)) stage, which was accompanied by enhanced Notch1 expression in T-cell progenitors. Consistently, constitutive activation of Notch1 drove the differentiation of Id1-expressing progenitors to the DN3 stage. Furthermore, we showed that Gfi1b levels decreased, whereas GATA3 levels increased in Id1 transgenic thymocytes. When overexpressed, GATA3 was able to upregulate Deltex1 transcription. Thus, T-cell commitment may be controlled by the interplay among E proteins, Gfi1b, and GATA3 transcription regulators, which influence Notch function through the expression of Deltex1. PMID- 19564411 TI - Receptor density is key to the alpha2/beta interferon differential activities. AB - Multiple type I interferons (IFN-alpha/beta) elicit Jak/Stat activation, rapid gene induction, and pleiotropic effects, such as differentiation, antiviral protection, and blocks in proliferation, which are dependent on the IFN subtype and the cellular context. To date, ligand- and receptor-specific molecular determinants underlying IFN-alpha/beta differential activities or potencies have been well characterized. To analyze cellular determinants that impact subtype specific potency, human fibrosarcoma U5A-derived clones, exhibiting a gradient of IFN sensitivity by virtue of increasing receptor levels, were monitored for Jak/Stat signaling, gene induction, cell cycle lengthening, and apoptosis. In cells with scarce receptors, IFN-beta was more potent than IFN-alpha2 in antiproliferative activity, while the two subtypes were equipotent in all other readouts. Conversely, in cells with abundant receptors, IFN-alpha2 matched or even surpassed IFN-beta in all readouts tested. Our results suggest that the differential activities of the IFN subtypes are dictated not only by the intrinsic ligand/receptor binding kinetics but also by the density of cell surface receptor components. PMID- 19564410 TI - Signal transduction cross talk mediated by Jun N-terminal kinase-interacting protein and insulin receptor substrate scaffold protein complexes. AB - Scaffold proteins have been established as important mediators of signal transduction specificity. The insulin receptor substrate (IRS) proteins represent a critical group of scaffold proteins that are required for signal transduction by the insulin receptor, including the activation of phosphatidylinositol 3 kinase. The c-Jun NH(2)-terminal kinase (JNK)-interacting proteins (JIPs) represent a different group of scaffold molecules that are implicated in the regulation of the JNK. These two signaling pathways are functionally linked because JNK can phosphorylate IRS1 on the negative regulatory site Ser-307. Here we demonstrate the physical association of these signaling pathways using a proteomic approach that identified insulin-regulated complexes of JIPs together with IRS scaffold proteins. Studies using mice with JIP scaffold protein defects confirm that the JIP1 and JIP2 proteins are required for normal glucose homeostasis. Together, these observations demonstrate that JIP proteins can influence insulin-stimulated signal transduction mediated by IRS proteins. PMID- 19564412 TI - The alternative TrkAIII splice variant targets the centrosome and promotes genetic instability. AB - The hypoxia-regulated alternative TrkAIII splice variant expressed by human neuroblastomas exhibits oncogenic potential, driven by in-frame exon 6 and 7 alternative splicing, leading to omission of the receptor extracellular immunoglobulin C(1) domain and several N-glycosylation sites. Here, we show that the TrkAIII oncogene promotes genetic instability by interacting with and exhibiting catalytic activity at the centrosome. This function depends upon intracellular TrkAIII accumulation and spontaneous interphase-restricted activation, in cytoplasmic tyrosine kinase (tk) domain orientation, predominantly within structures that closely associate with the fully assembled endoplasmic reticulum intermediate compartment and Golgi network. This facilitates TrkAIII tk mediated binding of gamma-tubulin, which is regulated by endogenous protein tyrosine phosphatases and geldanamycin-sensitive interaction with Hsp90, paving the way for TrkAIII recruitment to the centrosome. At the centrosome, TrkAIII differentially phosphorylates several centrosome-associated components, increases centrosome interaction with polo kinase 4, and decreases centrosome interaction with separase, the net results of which are centrosome amplification and increased genetic instability. The data characterize TrkAIII as a novel internal membrane-associated centrosome kinase, unveiling an important alternative mechanism to "classical" cell surface oncogenic receptor tk signaling through which stress-regulated alternative TrkAIII splicing influences the oncogenic process. PMID- 19564413 TI - Estrogen regulation of cyclin E2 requires cyclin D1 but not c-Myc. AB - During estrogen-induced proliferation, c-Myc and cyclin D1 initiate independent pathways that activate cyclin E1-Cdk2 by sequestration and/or downregulation of the CDK inhibitor p21(Waf1/Cip1), without significant increases in cyclin E1 protein levels. In contrast, cyclin E2 undergoes a marked increase in expression, which occurs within 9 to 12 h of estrogen treatment of antiestrogen-pretreated MCF-7 breast cancer cells. Both E cyclins are important to estrogen action, as small interfering RNA (siRNA)-mediated knockdown of either cyclin E1 or cyclin E2 attenuated estrogen-mediated proliferation. Inducible expression of cyclin D1 upregulated cyclin E2, while siRNA-mediated knockdown of cyclin D1 attenuated estrogen effects on cyclin E2. However, manipulation of c-Myc levels did not profoundly affect cyclin E2. Cyclin E2 induction by estrogen was accompanied by recruitment of E2F1 to the cyclin E1 and E2 promoters, and cyclin D1 induction was sufficient for E2F1 recruitment. siRNA-mediated knockdown of the chromatin remodelling factor CHD8 prevented cyclin E2 upregulation. Together, these data indicate that cyclin E2-Cdk2 activation by estrogen occurs via E2F- and CHD8 mediated transcription of cyclin E2 downstream of cyclin D1. This contrasts with the predominant regulation of cyclin E1-Cdk2 activity via CDK inhibitor association downstream of both c-Myc and cyclin D1 and indicates that cyclins E1 and E2 are not always coordinately regulated. PMID- 19564414 TI - The p107/E2F pathway regulates fibroblast growth factor 2 responsiveness in neural precursor cells. AB - We have previously shown that p107, a member of the retinoblastoma (Rb) cell cycle regulatory family, has a unique function in regulating the pool of neural precursor cells. As the pool of progenitors is regulated by a limiting supply of trophic factors, we asked if the Rb/E2F pathway may control the size of the progenitor population by regulating the levels of growth factors or their receptors. Here, we demonstrate that fibroblast growth factor 2 (FGF2) is aberrantly upregulated in the brains of animals lacking Rb family proteins and that the gene encoding the FGF2 ligand is directly regulated by p107 and E2F3. Chromatin immunoprecipitation assays demonstrated that E2F3 and p107 occupy E2F consensus sites on the FGF2 promoter in the context of native chromatin. To evaluate the physiological consequence of FGF2 deregulation in both p107 and E2F3 mutants, we measured neural progenitor responsiveness to growth factors. Our results demonstrate that E2F3 and p107 are each mediators of FGF2 growth factor responsiveness in neural progenitor cells. These results support a model whereby p107 regulates the pool of FGF-responsive progenitors by directly regulating FGF2 gene expression in vivo. By identifying novel roles for p107/E2F in regulating genes outside of the classical cell cycle machinery targets, we uncover a new mechanism whereby Rb/E2F mediates proliferation through regulating growth factor responsiveness. PMID- 19564415 TI - FOXO3a promotes tumor cell invasion through the induction of matrix metalloproteinases. AB - The role of the Forkhead transcription factor FOXO3a in processes that promote tumor metastasis is poorly defined. Here, we show that depletion of FOXO3a from cancer cells leads to decreased tumor size specifically due to attenuated invasive migration. During tumor progression, an increase in tumor mass is concomitant with serum deprivation prior to tumor angiogenesis. We show that nuclear retention of FOXO3a due to serum starvation results in greatly increased cancer cell invasion. Exploration of the mechanism by which FOXO3a promotes invasive migration revealed that it induces the expression of matrix metalloproteinase 9 (MMP-9) and MMP-13, both of which have been causally linked to the invasion and progression of numerous human solid tumors. Our results link Forkhead transcription factors to a previously unexplored function in cancer progression by promoting extracellular matrix degradation, allowing tumors to invade neighboring tissues and ultimately metastasize to distant organs. PMID- 19564417 TI - PU.1 can recruit BCL6 to DNA to repress gene expression in germinal center B cells. AB - BCL6 is a transcriptional repressor crucial for germinal center formation. BCL6 represses transcription by a variety of mechanisms by binding to specific DNA sequences or by recruitment to DNA by protein interactions. We found that BCL6 can inhibit activities of the immunoglobulin kappa (Igkappa) intron and 3' enhancers. At the Igkappa 3' enhancer, BCL6 repressed enhancer activity through the PU.1 binding site. We found that BCL6 physically interacted with PU.1 in vivo and in vitro, and the results of sequential chromatin immunoprecipitation assays and transient-expression assays suggested that BCL6 recruitment to the Igkappa and Iglambda 3' enhancers occurred via PU.1 interaction. By computational studies, we identified genes that are repressed in germinal center cells and whose promoters contain conserved PU.1 binding sites in mouse and human. We found that many of these promoters bound to both PU.1 and BCL6 in vivo. In addition, BCL6 knockdown resulted in increased expression of a subset of these genes, demonstrating that BCL6 is involved in their repression. The recruitment of BCL6 to promoter regions by PU.1 represents a new regulatory mechanism that expands the number of genes regulated by this important transcriptional repressor. PMID- 19564416 TI - Homodimerization controls the fibroblast growth factor 9 subfamily's receptor binding and heparan sulfate-dependent diffusion in the extracellular matrix. AB - Uncontrolled fibroblast growth factor (FGF) signaling can lead to human diseases, necessitating multiple layers of self-regulatory control mechanisms to keep its activity in check. Herein, we demonstrate that FGF9 and FGF20 ligands undergo a reversible homodimerization, occluding their key receptor binding sites. To test the role of dimerization in ligand autoinhibition, we introduced structure-based mutations into the dimer interfaces of FGF9 and FGF20. The mutations weakened the ability of the ligands to dimerize, effectively increasing the concentrations of monomeric ligands capable of binding and activating their cognate FGF receptor in vitro and in living cells. Interestingly, the monomeric ligands exhibit reduced heparin binding, resulting in their increased radii of heparan sulfate-dependent diffusion and biologic action, as evidenced by the wider dilation area of ex vivo lung cultures in response to implanted mutant FGF9-loaded beads. Hence, our data demonstrate that homodimerization autoregulates FGF9 and FGF20's receptor binding and concentration gradients in the extracellular matrix. Our study is the first to implicate ligand dimerization as an autoregulatory mechanism for growth factor bioactivity and sets the stage for engineering modified FGF9 subfamily ligands, with desired activity for use in both basic and translational research. PMID- 19564418 TI - The mTORC2 complex regulates terminal differentiation of C2C12 myoblasts. AB - Rapamycin, a selective inhibitor of mTORC1 signaling, blocks terminal myoblast differentiation. We found that downregulation of rictor, a component of the mTORC2 complex, but not downregulation of raptor, a component of the mTORC1 complex, prevented terminal differentiation (fusion) of C2C12 myoblasts. Both rapamycin and rictor downregulation suppressed the phosphorylation of AKT(S(473)), and rapamycin treatment of C2C12 myoblasts disrupted the mTORC2 complex. Importantly, downregulation of rictor inhibited TORC2 signaling without inhibiting mTORC1 signaling, suggesting that inhibition of mTORC1 by rapamycin may not be the cause of arrested differentiation. In support of this, expression of a phosphomimetic mutant AKT(S473D) in rictor-deficient cells rescued myoblast fusion even in the presence of rapamycin. mTORC2 signaling to AKT appears necessary for downregulation of the Rho-associated kinase (ROCK1) that occurs during myogenic differentiation. Rapamycin treatment prevented ROCK1 inactivation during differentiation, while suppression of ROCK1 activity during differentiation and myoblast fusion was restored through expression of AKT(S473D), even in the presence of rapamycin. Further, the ROCK inhibitor Y 27632 restored terminal differentiation in rapamycin-treated myoblasts. These results provide the first evidence of a specific role for mTORC2 signaling in terminal myogenic differentiation. PMID- 19564419 TI - Alpha interferon induces long-lasting refractoriness of JAK-STAT signaling in the mouse liver through induction of USP18/UBP43. AB - Recombinant alpha interferon (IFN-alpha) is used for the treatment of viral hepatitis and some forms of cancer. During these therapies IFN-alpha is injected once daily or every second day for several months. Recently, the long-acting pegylated IFN-alpha (pegIFN-alpha) has replaced standard IFN-alpha in therapies of chronic hepatitis C because it is more effective, supposedly by inducing a long-lasting activation of IFN signaling pathways. IFN signaling in cultured cells, however, becomes refractory within hours, and little is known about the pharmacodynamic effects of continuously high IFN-alpha serum concentrations. To investigate the behavior of the IFN system in vivo, we repeatedly injected mice with IFN-alpha and analyzed its effects in the liver. Within hours after the first injection, IFN-alpha signaling became refractory to further stimulation. The negative regulator SOCS1 was rapidly upregulated and likely responsible for early termination of IFN-alpha signaling. For long-lasting refractoriness, neither SOCS1 nor SOCS3 were instrumental. Instead, we identified the inhibitor USP18/UBP43 as the key mediator. Our results indicate that the current therapeutic practice using long-lasting pegIFN-alpha is not well adapted to the intrinsic properties of the IFN system. Targeting USP18 expression may allow to exploit the full therapeutic potential of recombinant IFN-alpha. PMID- 19564420 TI - Sterol regulatory element binding protein 1a regulates hepatic fatty acid partitioning by activating acetyl coenzyme A carboxylase 2. AB - We generated a line of mice in which sterol regulatory element binding protein 1a (SREBP-1a) was specifically inactivated by insertional mutagenesis. Homozygous mutant mice were completely viable despite expressing SREBP-1a mRNA below 5% of normal, and there were minimal effects on expression of either SREBP-1c or -2. Microarray expression studies in liver, where SREBP-1a mRNA is 1/10 the level of the highly similar SREBP-1c, demonstrated that only a few genes were affected. The only downregulated genes directly linked to lipid metabolism were Srebf1 (which encodes SREBP-1) and Acacb (which encodes acetyl coenzyme A [acetyl-CoA] carboxylase 2 [ACC2], a critical regulator of fatty acyl-CoA partitioning between cytosol and mitochondria). ACC2 regulation is particularly important during food restriction. Similar to Acacb knockout mice, SREBP-1a-deficient mice have lower hepatic triglycerides and higher serum ketones during fasting than wild-type mice. SREBP-1a and -1c have identical DNA binding and dimerization domains; thus, the failure of the more abundant SREBP-1c to substitute for activating hepatic ACC2 must relate to more efficient recruitment of transcriptional coactivators to the more potent SREBP-1a activation domain. Our chromatin immunoprecipitation results support this hypothesis. PMID- 19564421 TI - Time-dependent activation of Phox2a by the cyclic AMP pathway modulates onset and duration of p27Kip1 transcription. AB - In noradrenergic progenitors, Phox2a mediates cell cycle exit and neuronal differentiation by inducing p27(Kip1) transcription in response to activation of the cyclic AMP (cAMP) pathway. The mechanism of cAMP-mediated activation of Phox2a is unknown. We identified a cluster of phosphoserine-proline sites in Phox2a by mass spectrometry. Ser206 appeared to be the most prominent phosphorylation site. A phospho-Ser206 Phox2a antibody detected dephosphorylation of Phox2a that was dependent on activation of the cAMP pathway, which occurred prior to neuronal differentiation of noradrenergic CAD cells. Employing serine-to alanine and serine-to-aspartic acid Phox2a substitution mutants expressed in inducible CAD cell lines, we demonstrated that the transcriptional activity of Phox2a is regulated by two sequential cAMP-dependent events: first, cAMP signaling promotes dephosphorylation of Phox2a in at least one site, Ser206, thereby allowing Phox2a to bind DNA and initiate p27(Kip1) transcription; second, following dephosphorylation of the phosphoserine cluster (Ser202 and Ser208), Phox2a becomes phosphorylated by protein kinase A (PKA) on Ser153, which prevents association of Phox2a with DNA and terminates p27(Kip1) transcription. This represents a novel mechanism by which the same stimulus, cAMP signaling, first activates Phox2a by dephosphorylation of Ser206 and then, after a built-in delay, inactivates Phox2a via PKA-dependent phosphorylation of Ser153, thereby modulating onset and duration of p27(Kip1) transcription. PMID- 19564422 TI - Developmental control of CaV1.2 L-type calcium channel splicing by Fox proteins. AB - CaV1.2 voltage-gated calcium channels play critical roles in the control of membrane excitability, gene expression, and muscle contraction. These channels show diverse functional properties generated by alternative splicing at multiple sites within the CaV1.2 pre-mRNA. The molecular mechanisms controlling this splicing are not understood. We find that two exons in the CaV1.2 channel are controlled in part by members of the Fox family of splicing regulators. Exons 9* and 33 confer distinct electrophysiological properties on the channel and show opposite patterns of regulation during cortical development, with exon 9* progressively decreasing its inclusion in the CaV1.2 mRNA over time and exon 33 progressively increasing. Both exons contain Fox protein binding elements within their adjacent introns, and Fox protein expression is induced in cortical neurons in parallel with the changes in CaV1.2 splicing. We show that knocking down expression of Fox proteins in tissue culture cells has opposite effects on exons 9* and 33. The loss of Fox protein increases exon 9* splicing and decreases exon 33, as predicted by the positions of the Fox binding elements and by the pattern of splicing in development. Conversely, overexpression of Fox1 and Fox2 proteins represses exon 9* and enhances exon 33 splicing in the endogenous CaV1.2 mRNA. These effects of Fox proteins on exons 9* and 33 can be recapitulated in transfected minigene reporters. Both the repressive and the enhancing effects of Fox proteins are dependent on the Fox binding elements within and adjacent to the target exons, indicating that the Fox proteins are directly regulating both exons. These results demonstrate that the Fox protein family is playing a key role in tuning the properties of CaV1.2 calcium channels during neuronal development. PMID- 19564423 TI - Double-bundle anterior cruciate ligament reconstruction: a comprehensive kinematic study using navigation. AB - BACKGROUND: Single-bundle anterior cruciate ligament reconstruction seems to be insufficient to control a combined rotatory load of internal and valgus torque, whereas anatomical double-bundle reconstruction might produce a better biomechanical outcome, especially during rotatory loads. HYPOTHESIS: The addition of the posterolateral bundle to the anteromedial bundle, in an in vivo double bundle computer-assisted anterior cruciate ligament reconstruction, is able to reduce the internal rotation of the tibia at different degrees of flexion. STUDY DESIGN: Controlled laboratory study. METHODS: Ten consecutive anterior cruciate ligament reconstruction procedures were performed in male patients using double bundle gracilis and semitendinosus tendon graft with the 2.0 OrthoPilot anterior cruciate ligament navigation system. Anteroposterior displacement at 30 degrees , 60 degrees , and 90 degrees as well as internal and external rotation at 0 degrees , 15 degrees , 30 degrees , 45 degrees , 60 degrees , and 90 degrees of knee flexion were evaluated before reconstruction, after fixation of the anteromedial bundle, and then after fixation of the posterolateral bundle. RESULTS: Fixation of the anteromedial bundle significantly (P < .05) reduced the anteroposterior displacement at 30 degrees , 60 degrees , and 90 degrees of knee flexion; the internal rotation at 15 degrees , 30 degrees , 45 degrees , and 60 degrees ; and the external rotation at 0 degrees , 30 degrees , 60 degrees , and 90 degrees . The addition of the posterolateral bundle to the anteromedial bundle did not significantly reduce internal and external rotation of the tibia at degrees of flexion measured (P > .05). CONCLUSION: The hypothesis that the addition of the posterolateral bundle to the anteromedial bundle is able to reduce internal rotation of the tibia, minimizing the pivot-shift phenomenon, was not confirmed. CLINICAL RELEVANCE: The effective role of the anatomical double bundle procedure in better restoring knee kinematics and allowing better clinical outcomes should be questioned in an in vivo model of anterior cruciate ligament reconstruction using doubled semitendinosus and gracilis graft. PMID- 19564424 TI - Another story of arginines in voltage sensing: the role of phosphoinositides in coupling voltage sensing to enzyme activity. PMID- 19564425 TI - Coupling between the voltage-sensing and phosphatase domains of Ci-VSP. AB - The Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) shares high homology with the phosphatidylinositol phosphatase enzyme known as PTEN (phosphatase and tensin homologue deleted on chromosome 10). We have taken advantage of the similarity between these proteins to inquire about the coupling between the voltage sensing and the phosphatase domains in Ci-VSP. Recently, it was shown that four basic residues (R11, K13, R14, and R15) in PTEN are critical for its binding onto the membrane, required for its catalytic activity. Ci-VSP has three of the basic residues of PTEN. Here, we show that when R253 and R254 (which are the homologues of R14 and R15 in PTEN) are mutated to alanines in Ci VSP, phosphatase activity is disrupted, as revealed by a lack of effect on the ionic currents of KCNQ2/3, where current decrease is a measure of phosphatase activity. The enzymatic activity was not rescued by the introduction of lysines, indicating that the binding is an arginine-specific interaction between the phosphatase binding domain and the membrane, presumably through the phosphate groups of the phospholipids. We also found that the kinetics and steady-state voltage dependence of the S4 segment movement are affected when the arginines are not present, indicating that the interaction of R253 and R254 with the membrane, required for the catalytic action of the phosphatase, restricts the movement of the voltage sensor. PMID- 19564429 TI - Asymptomatic large unruptured left ventricular pseudoaneurysm. PMID- 19564427 TI - A structural model for K2P potassium channels based on 23 pairs of interacting sites and continuum electrostatics. AB - K(2P)O, the two-pore domain potassium background channel that determines cardiac rhythm in Drosophila melanogaster, and its homologues that establish excitable membrane activity in mammals are of unknown structure. K(2P) subunits have two pore domains flanked by transmembrane (TM) spans: TM1-P1-TM2-TM3-P2-TM4. To establish spatial relationships in K(2P)O, we identified pairs of sites that display electrostatic compensation. Channels silenced by the addition of a charge in pore loop 1 (P1) or P2 were restored to function by countercharges at specific second sites. A three-dimensional homology model was determined using the crystal structure of K(V)1.2, effects of K(2P)O mutations to establish alignment, and compensatory charge-charge pairs. The model was refined and validated by continuum electrostatic free energy calculations and covalent linkage of introduced cysteines. K(2P) channels use two subunits arranged so that the P1 and P2 loops contribute to one pore, identical P loops face each other diagonally across the pore, and the channel complex has bilateral symmetry with a fourfold symmetric selectivity filter. PMID- 19564426 TI - Effects of inserting fluorescent proteins into the alpha1S II-III loop: insights into excitation-contraction coupling. AB - In skeletal muscle, intermolecular communication between the 1,4-dihydropyridine receptor (DHPR) and RYR1 is bidirectional: orthograde coupling (skeletal excitation-contraction coupling) is observed as depolarization-induced Ca(2+) release via RYR1, and retrograde coupling is manifested by increased L-type Ca(2+) current via DHPR. A critical domain (residues 720-765) of the DHPR alpha(1S) II-III loop plays an important but poorly understood role in bidirectional coupling with RYR1. In this study, we examine the consequences of fluorescent protein insertion into different positions within the alpha(1S) II III loop. In four constructs, a cyan fluorescent protein (CFP)-yellow fluorescent protein (YFP) tandem was introduced in place of residues 672-685 (the peptide A region). All four constructs supported efficient bidirectional coupling as determined by the measurement of L-type current and myoplasmic Ca(2+) transients. In contrast, insertion of a CFP-YFP tandem within the N-terminal portion of the critical domain (between residues 726 and 727) abolished bidirectional signaling. Bidirectional coupling was partially preserved when only a single YFP was inserted between residues 726 and 727. However, insertion of YFP near the C terminal boundary of the critical domain (between residues 760 and 761) or in the conserved C-terminal portion of the alpha(1S) II-III loop (between residues 785 and 786) eliminated bidirectional coupling. None of the fluorescent protein insertions, even those that interfered with signaling, significantly altered membrane expression or targeting. Thus, bidirectional signaling is ablated by insertions at two different sites in the C-terminal portion of the alpha(1S) II III loop. Significantly, our results indicate that the conserved portion of the alpha(1S) II-III loop C terminal to the critical domain plays an important role in bidirectional coupling either by conveying conformational changes to the critical domain from other regions of the DHPR or by serving as a site of interaction with other junctional proteins such as RYR1. PMID- 19564430 TI - Rare case of blunt chest trauma induced left main and LAD dissection in association with anomalous RCA origin. PMID- 19564431 TI - Coronary heart disease epidemics. PMID- 19564432 TI - Cardiovascular manifestations of HIV infection. PMID- 19564433 TI - Steroid-responsive sterile inflammation after transradial cardiac catheterisation using a sheath with hydrophilic coating. PMID- 19564434 TI - Evaluating the capability and cost of a mass influenza and pneumococcal vaccination clinic via computer simulation. AB - OBJECTIVE: To determine if a mass influenza/pneumococcal vaccination clinic could vaccinate 15,000 clients in 17 h; optimize personnel configuration to maximize number of clients vaccinated; and estimate costs (opportunity and clinic) and revenue. METHOD: The author used a discrete event simulation model to estimate the throughput of the vaccination clinic as the number of clients (arrival intensity) increased and as staff members were reassigned to different workflows. We represented workflows for 3 client types: "Medicare,'' "Special,'' and "Cash,'' where "Special'' designates Medicare clients who needed assistance moving through the clinic. The costs of supplies, staff sal-aries, and client waiting time were included in the model. We compared the "original'' model based on the staffing and performance of an actual clinic to an ;;optimized'' model in which staff were reassigned to optimize number of clients vaccinated. RESULTS: A maximum of 13,138 and 15,094 clients in the original and optimized models, respectively, were vaccinated. At the original arrival rate (8300 clients vaccinated in 17 h), supplies cost about $191,000 and were the most expensive component of the clinic operation in both models. However, as the arrival intensity increased to 140%, the "Medicare'' client opportunity cost increased from $23,887 and $21,474 to $743,510 and $740,760 for the simulated original and optimized models, respectively. CONCLUSION: The clinic could reach their target of 15,000 vaccinees with 2 fewer staff members by rearranging staff assignments from "Special" to "Medicare'' and "Cash'' stations. Computer simulation can help public health officials determine the most efficient use of staff, machinery, supplies, and time. PMID- 19564435 TI - Pediatricians' knowledge of, experience with, and comfort levels for cochlear implants in children. AB - PURPOSE: Pediatricians are vital to early hearing detection and intervention programs (EHDIPs) and influence families' decisions and compliance with recommendations for their children having hearing loss. Pediatricians need current, accurate knowledge about diagnostics and treatments including cochlear implants (CIs). This study assessed pediatricians' knowledge about, experience with, and comfort levels for CIs in children. METHOD: Questionnaires were mailed to a total of 220 pediatricians in 9 cities in Alabama. RESULTS: Nine questionnaires were undeliverable; of the remaining 211 surveys, 29 were returned (for a response rate of 13.7%), and 26 of those were usable. The results were consistent with other studies. All respondents focused on pediatrics; most were medical homes experienced with children having sensorineural hearing loss. Less than half had counseled families about CIs but had patients using them. Over half had not recommended CIs in 5 years. Several were unsure about current EHDIP goals and CIs in children. Over half were completely uncomfortable determining children's CI candidacy but were comfortable referring them to specialists. About a third were completely uncomfortable discussing CIs with and counseling families about them; most were comfortable with CIs being used in children over other habilitative options. CONCLUSIONS: These pediatricians demonstrated deficits in current, accurate knowledge about EHDIPs and CIs in children. Audiologists should help them obtain needed information. PMID- 19564436 TI - Comparison of acoustic and kinematic approaches to measuring utterance-level speech variability. AB - PURPOSE: The spatiotemporal index (STI) is one measure of variability. As currently implemented, kinematic data are used, requiring equipment that cannot be used with some patient groups or in scanners. An experiment is reported that addressed whether STI can be extended to an audio measure of sound pressure of the speech envelope over time that did not need specialized equipment. METHOD: STI indices of variability were obtained from lip track (L-STI) and amplitude envelope (E-STI) signals. These measures were made concurrent while either fluent speakers or speakers who stutter repeated "Buy Bobby a puppy" 20 times. RESULTS: L-STI and E-STI correlated significantly. STI decreased with age for both L-STI and E-STI. E-STI scores and L-STI scores discriminated successfully between fluent speakers and speakers who stutter. CONCLUSION: The amplitude-envelope-over time signal can be used to obtain an STI score. This STI score can be used in situations where lip movement STI scores are precluded. PMID- 19564437 TI - Directional effects on infants and young children in real life: implications for amplification. AB - PURPOSE: This study examined the head orientation of young children in naturalistic settings and the acoustics of their everyday environments for quantifying the potential effects of directionality. METHOD: Twenty-seven children (11 with normal hearing, 16 with impaired hearing) between 11 and 78 months of age were video recorded in naturalistic settings for analyses of head orientation. Reports on daily activities were obtained from caregivers. The effect of directionality in different environments was quantified by measuring the Speech Transmission Index (STI; H. J. M. Steeneken & T. Houtgast, 1980). RESULTS: Averaged across 4 scenarios, children looked in the direction of a talker for 40% of the time when speech was present. Head orientation was not affected by age or hearing status. The STI measurements revealed a directional advantage of 3 dB when a child looked at a talker but a deficit of 2.8 dB when the talker was sideways or behind the child. The overall directional effect in real life was between -0.4 and 0.2 dB. CONCLUSIONS: The findings suggest that directional microphones in personal hearing devices for young children are not detrimental and have much potential for benefits in real life. The benefits may be enhanced by fitting directionality early and by counseling caregivers on ways to maximize benefits in everyday situations. PMID- 19564438 TI - Development and perceptual evaluation of amplitude-based F0 control in electrolarynx speech. AB - PURPOSE: Current electrolarynx (EL) devices produce a mechanical speech quality that has been largely attributed to the lack of natural fundamental frequency (F0) variation. In order to improve the quality of EL speech, in the present study the authors aimed to develop and evaluate an automatic F0 control scheme, in which F0 was modulated based on variations in the root-mean-square (RMS) amplitude of the EL speech signal. METHOD: Recordings of declarative sentences produced by 2 male participants before and after total laryngectomy were used to develop procedures for calculating F0 contours for EL speech. Specifically, the positive linear relationship between F0 and RMS amplitude observed in pre laryngectomy speech was used as the basis for generating an F0 contour based on the amplitude variation of EL speech. An analysis-by-synthesis approach was used to modify the F0 contour, and a perceptual experiment was conducted to examine its impact on the quality of the EL speech. RESULTS: The results of perceptual experiments showed that modulating the F0 of EL speech using a linear relationship between amplitude and frequency made it significantly more natural sounding than EL speech with constant F0. CONCLUSIONS: The current study provides preliminary support for amplitude-based control of F0 in EL speech. PMID- 19564440 TI - Do we do literacy? PMID- 19564439 TI - The effects of Fast ForWord Language on the phonemic awareness and reading skills of school-age children with language impairments and poor reading skills. AB - PURPOSE: To examine the efficacy of Fast ForWord Language (FFW-L) and 2 other interventions for improving the phonemic awareness and reading skills of children with specific language impairment with concurrent poor reading skills. METHOD: A total of 103 children (age 6;0 to 8;11 [years;months]) with language impairment and poor reading skills participated. The children received either FFW-L computerized intervention, a computer-assisted language intervention (CALI), an individualized language intervention (ILI), or an attention control (AC) computer program. RESULTS: The children in the FFW-L, CALI, and ILI conditions made significantly greater gains in blending sounds in words compared with the AC group at immediate posttest. Long-term gains 6 months after treatment were not significant but yielded a medium effect size for blending sounds in words. None of the interventions led to significant changes in reading skills. CONCLUSION: The improvement in phonemic awareness, but not reading, in the FFW-L, CALI, and ILI interventions limits their use with children who have language impairment and poor reading skills. Similar results across treatment conditions suggest that acoustically modified speech was not a necessary component for improving phonemic awareness. PMID- 19564441 TI - A survey of university professors teaching speech sound disorders: nonspeech oral motor exercises and other topics. AB - PURPOSE: The purpose of this article was to obtain and organize information from instructors who teach course work on the subject of children's speech sound disorders (SSD) regarding their use of teaching resources, involvement in students' clinical practica, and intervention approaches presented to students. Instructors also reported if they taught students to use nonspeech oral motor exercises (NSOMEs) to remediate children's SSD. METHOD: A questionnaire was mailed to 236 speech-language pathology preprofessional programs in the United States that are accredited by the Council of Academic Accreditation. RESULTS: Ninety-one questionnaires (39%) were returned. Participants reported that they provided their students with information on a variety of intervention approaches for SSD and typically used professional journals and textbooks for current information. Sixty-eight (75%) paticipants reported that they did not teach their students to use NSOMEs. Forty-seven (52%) of the instructors supervised students in clinical practicum serving children with SSD and perceived that academic course work and practicum experiences influenced their students' implementation of intervention for children with SSD. CONCLUSION: The instructors reported that they taught their students a variety of intervention techniques for children with SSD, although most did not teach the use of NSOMEs. These results contrast with previous research indicating that many speech-language pathologists use NSOMEs to improve children's speech (G.L. Lof & M.M. Watson, 2008). PMID- 19564442 TI - Morphology and literacy: getting our heads in the game. AB - PURPOSE: This prologue introduces the clinical forum, briefly discusses the importance of morphology in literacy, and informs the reader of the scope of the included articles. METHOD: The concept of morphology is reviewed, contributing authors are introduced, and a brief summary of each of the 5 forum articles is provided. CONCLUSION: The studies in this forum investigated different morphological skills in a variety of contexts, at a variety of grade levels, and from a variety of perspectives, enabling the reader to learn more about morphological awareness developmentally, linguistically, and clinically. PMID- 19564443 TI - The influence of morphological awareness on the literacy development of first grade children. AB - PURPOSE: The purpose of this study was twofold. First, we investigated whether first-grade children evidenced morphological awareness and whether they used their knowledge of morphological relations to guide their spelling. Second, we sought to determine whether children's morphological awareness abilities were predictive of their performance on word-level reading and spelling measures. METHOD: At the beginning of the academic school year, 43 first-grade children were administered an oral morphological awareness production task, a series of single-word morphological spelling tasks, and a battery of language and literacy tasks. RESULTS: The first-grade children were able to generate words reflecting morphological relations before they received explicit instruction regarding morphological relations between words. In addition, the children used morphological information to guide their spelling of single words, as evidenced by a difference in patterns of spellings between 1- and 2-morpheme words. Regression analyses revealed that the children's performance on the oral morphological production task explained unique variance on their reading and spelling measures above and beyond the variance that was accounted for by phonological awareness. CONCLUSION: Children as young as first graders evidenced morphological awareness, and morphological awareness influenced the children's literacy development. Theoretical implications of the findings are discussed. PMID- 19564444 TI - Morphological awareness skills of fourth-grade African American students. AB - PURPOSE: We examined the morphological awareness skills of fourth-grade African American children and the association between degree of African American English (AAE) use and performance on written measures of morphological awareness. Additional purposes were to determine whether performance on the morphological awareness tasks (a) was affected by the transparency of morphologically related words and the type of task administered, (b) was associated with other literacy and literacy-related skills, and (c) explained unique variance on these latter abilities. METHOD: Thirty fourth-grade African American children from low-income backgrounds were administered 2 morphological awareness tasks and completed norm referenced measures of word-level reading, reading comprehension, spelling, phonemic awareness, and receptive vocabulary. RESULTS: The degree of AAE use was not associated with students' performance on the morphological awareness tasks. On these tasks, significantly higher scores were obtained on items that represented a transparent relationship between a base word and its derived form. The students' performance on the morphological awareness tasks was significantly and moderately related to their performance on the word-level reading, spelling, and receptive vocabulary measures. Morphological awareness scores explained significant unique variance on measures of word-level reading and spelling, above that predicted by performance on measures of phonemic awareness and vocabulary. CONCLUSION: As shown in previous investigations of Caucasian children's morphological awareness skills, fourth-grade African American students' morphological awareness abilities are associated with select language and literacy skills. Professionals should capitalize on students' intact capabilities in morphological awareness during literacy instruction in an effort to maximize language and literacy performance for African American students. PMID- 19564445 TI - Teaching students with reading difficulties to be close readers: a feasibility study. AB - PURPOSE: This article describes a program that was designed to help upper elementary students read and understand words as they read texts independently. As a first step in helping middle-to-upper elementary children with mild-to moderate language and/or reading difficulties engage in textual analysis during reading, the Close Reading program combines instruction in morphological-analysis and context-analysis strategies with guided experiences applying these strategies during reading. METHOD: To carry out an initial feasibility study of the program, we conducted 3 case studies using standardized pretest and posttest measures of language and reading skills and experimental progress monitoring measures administered before, during, and after instruction. Three fourth-grade girls participated in the 12-week program. RESULTS: All 3 students showed improved word reading and comprehension with small to large effect sizes on standardized and experimental measures. Patterns of improvement reflected the initial strengths and weaknesses of the students' reading and language skills. CONCLUSION: The results suggest that further experimental investigation of this program is warranted. Instruction in morphological-analysis strategies with guided practice during reading holds promise as a way to improve word reading and comprehension for struggling readers in the middle-to-upper elementary years. PMID- 19564446 TI - Epilogue: Morphology and literacy: getting (and keeping) our heads in the game. PMID- 19564447 TI - Development and applications of a broad-coverage, TR-FRET-based kinase binding assay platform. AB - The expansion of kinase assay technologies over the past decade has mirrored the growing interest in kinases as drug targets. As a result, there is no shortage of convenient, fluorescence-based methods available to assay targets that span the kinome. The authors recently reported on the development of a non-activity-based assay to characterize kinase inhibitors that depended on displacement of an Alexa Fluor 647 conjugate of staurosporine (a "tracer") from a particular kinase. Kinase inhibitors were characterized by a change in fluorescence lifetime of the tracer when it was bound to a kinase relative to when it was displaced by an inhibitor. Here, the authors report on improvements to this strategy by reconfiguring the assay in a time-resolved fluorescence resonance energy transfer (TR-FRET) format that simplifies instrumentation requirements and allows for the use of a substantially lower concentration of kinase than was required in the fluorescence-lifetime-based format. The authors use this new assay to demonstrate several aspects of the binding assay format that are advantageous relative to traditional activity-based assays. The TR-FRET binding format facilitates the assay of compounds against low-activity kinases, allows for the characterization of type II kinase inhibitors either using nonactivated kinases or by monitoring compound potency over time, and ensures that the signal being detected is specific to the kinase of interest and not a contaminating kinase. PMID- 19564448 TI - Overweight prevention in pediatric primary care: a needs assessment of an urban racial/ethnic minority population. AB - The authors studied the prevalence of overweight-related behaviors in an urban clinic population, parents' perceived willingness to change, and identified potential gaps in nutrition and physical activity promotion. A total of 324 parents of children aged 3 to 13 years were surveyed. Clinical heights and weights were used to calculate body mass index (BMI). Of the 324 children in the study, 55% were black and 28% were Hispanic. Approximately 151 (47%) children had a BMI > or =85th percentile, and overweight-related behaviors, such as TV viewing, were highly prevalent. Overall, parents reported a need for counseling to help their children eat healthier and be more active and seemed willing to make behavior changes in these areas. However, their willingness to change appeared lowest in areas that may improve their child's weight status such as decreasing sedentary time and portion sizes. Overweight prevention efforts in primary care should include strategies to help clinicians negotiate behavior change with families. PMID- 19564449 TI - Adding omalizumab to the therapy of adolescents with persistent uncontrolled moderate--severe allergic asthma. AB - OBJECTIVE: This study aimed to evaluate the effectiveness of omalizumab among adolescents with moderate-severe allergic asthma inadequately controlled with inhaled corticosteroids. PATIENTS AND METHODS: Data from patients 12 to 17 years of age were pooled from 5 placebo-controlled registration trials of omalizumab. Impact on asthma control was assessed by need for rescue bursts of oral corticosteroids, lung function, symptom scores, and unscheduled office visits. RESULTS: In adolescents (n = 146), addition of omalizumab decreased mean number of rescue bursts (0.3 vs 0.9) versus placebo; relative risk 0.47 (95% confidence interval [CI], 0.22-0.99; P = .047). At study conclusion, mean forced expiratory volume in 1 second increased 268 mL (13.8%) in omalizumab-treated subjects versus 98 mL (5.5%) for placebo (least squares mean treatment difference 146 mL [95% CI, 19.4-272.6; P = .024]). Omalizumab significantly improved asthma symptom scores and reduced unscheduled office visits. CONCLUSION: Omalizumab added to baseline therapy improves measures of asthma control in adolescents with persistent moderate-severe allergic asthma. PMID- 19564450 TI - Practice patterns of pediatric emergency medicine physicians caring for young febrile infants. AB - The authors conducted a chart review of all febrile infants between 28 and 90 days of age who presented to the emergency department (ED) between December 1 and March 31 during 2004-2006. The objectives of the study were to describe the practice patterns of pediatric ED physicians caring for these infants and to determine whether the evaluation and management of these infants differed based on their age at presentation. Two groups were compared-infants aged 28 to 59 days (n = 79) and infants aged 60 to 90 days (n = 88). As compared with the younger age group, infants in the older age group had fewer complete blood cell counts (relative risk, RR = 3.57; 95% confidence interval [CI], 2.15-5.95), fewer blood cultures (RR = 3.38; 95% CI, 1.99-5.74), fewer urine cultures (RR = 3.83; 95% CI, 1.81-8.13), and fewer cerebrospinal fluid cultures (RR = 2.56; 95% CI, 1.94 3.40). Overall, there was poor adherence to current guidelines for the diagnostic evaluation of young febrile infants. PMID- 19564451 TI - Inference of the impact of insertion sequence (IS) elements on bacterial genome diversification through analysis of small-size structural polymorphisms in Escherichia coli O157 genomes. AB - Mobile genetic elements play important roles in the evolution and diversification of bacterial genomes. In enterohemorrhagic Escherichia coli O157, a major factor that affects genomic diversity is prophages, which generate most of the large size structural polymorphisms (LSSPs) observed in O157 genomes. Here, we describe the results of a systematic analysis of numerous small-size structural polymorphisms (SSSPs) that were detected by comparing the genomes of eight clinical isolates with a sequenced strain, O157 Sakai. Most of the SSSPs were generated by genetic events associated with only two insertion sequence (IS) elements, IS629 and ISEc8, and a number of genes that were inactivated or deleted by these events were identified. Simple excisions of IS629 and small deletions (footprints) formed by the excision of IS629, both of which are rarely described in bacteria, were also detected. In addition, the distribution of IS elements was highly biased toward prophages, prophage-like integrative elements, and plasmids. Based on these and our previous results, we conclude that, in addition to prophages, these two IS elements are major contributors to the genomic diversification of O157 strains and that LSSPs have been generated mainly by bacteriophages and SSSPs by IS elements. We also suggest that IS elements possibly play a role in the inactivation and immobilization of incoming phages and plasmids. Taken together, our results reveal the true impact of IS elements on the diversification of bacterial genomes and highlight their novel role in genome evolution. PMID- 19564452 TI - mGene: accurate SVM-based gene finding with an application to nematode genomes. AB - We present a highly accurate gene-prediction system for eukaryotic genomes, called mGene. It combines in an unprecedented manner the flexibility of generalized hidden Markov models (gHMMs) with the predictive power of modern machine learning methods, such as Support Vector Machines (SVMs). Its excellent performance was proved in an objective competition based on the genome of the nematode Caenorhabditis elegans. Considering the average of sensitivity and specificity, the developmental version of mGene exhibited the best prediction performance on nucleotide, exon, and transcript level for ab initio and multiple genome gene-prediction tasks. The fully developed version shows superior performance in 10 out of 12 evaluation criteria compared with the other participating gene finders, including Fgenesh++ and Augustus. An in-depth analysis of mGene's genome-wide predictions revealed that approximately 2200 predicted genes were not contained in the current genome annotation. Testing a subset of 57 of these genes by RT-PCR and sequencing, we confirmed expression for 24 (42%) of them. mGene missed 300 annotated genes, out of which 205 were unconfirmed. RT-PCR testing of 24 of these genes resulted in a success rate of merely 8%. These findings suggest that even the gene catalog of a well-studied organism such as C. elegans can be substantially improved by mGene's predictions. We also provide gene predictions for the four nematodes C. briggsae, C. brenneri, C. japonica, and C. remanei. Comparing the resulting proteomes among these organisms and to the known protein universe, we identified many species-specific gene inventions. In a quality assessment of several available annotations for these genomes, we find that mGene's predictions are most accurate. PMID- 19564453 TI - New users of metformin are at low risk of incident cancer: a cohort study among people with type 2 diabetes. AB - OBJECTIVE: The antidiabetic properties of metformin are mediated through its ability to activate the AMP-activated protein kinase (AMPK). Activation of AMPK can suppress tumor formation and inhibit cell growth in addition to lowering blood glucose levels. We tested the hypothesis that metformin reduces the risk of cancer in people with type 2 diabetes. RESEARCH DESIGN AND METHODS: In an observational cohort study using record-linkage databases and based in Tayside, Scotland, U.K., we identified people with type 2 diabetes who were new users of metformin in 1994-2003. We also identified a set of diabetic comparators, individually matched to the metformin users by year of diabetes diagnosis, who had never used metformin. In a survival analysis we calculated hazard ratios for diagnosis of cancer, adjusted for baseline characteristics of the two groups using Cox regression. RESULTS: Cancer was diagnosed among 7.3% of 4,085 metformin users compared with 11.6% of 4,085 comparators, with median times to cancer of 3.5 and 2.6 years, respectively (P < 0.001). The unadjusted hazard ratio (95% CI) for cancer was 0.46 (0.40-0.53). After adjusting for sex, age, BMI, A1C, deprivation, smoking, and other drug use, there was still a significantly reduced risk of cancer associated with metformin: 0.63 (0.53-0.75). CONCLUSIONS: These results suggest that metformin use may be associated with a reduced risk of cancer. A randomized trial is needed to assess whether metformin is protective in a population at high risk for cancer. PMID- 19564454 TI - Maturity-onset diabetes of the young in children with incidental hyperglycemia: a multicenter Italian study of 172 families. AB - OBJECTIVE: To investigate the prevalence of maturity-onset diabetes of the young (MODY) in Italian children with incidental hyperglycemia. RESEARCH DESIGN AND METHODS: Among 748 subjects age 1-18 years with incidental hyperglycemia, minimal diagnostic criteria for MODY were met by 172 families. Mutational analyses of the glucokinase (GCK) and hepatocyte nuclear factor 1alpha (HNF1A) genes were performed. RESULTS: We identified 85 GCK gene mutations in 109 probands and 10 HNF1A mutations in 12 probands. In GCK patients, the median neonatal weight and age at the first evaluation were lower than those found in patients with HNF1A mutations. Median fasting plasma glucose and impaired fasting glucose/impaired glucose tolerance frequency after oral glucose tolerance testing were higher in GCK patients, who also showed a lower frequency of diabetes than HNF1A patients. CONCLUSIONS: GCK mutations are the prevailing cause of MODY (63.4%) when the index case is recruited in Italian children with incidental hyperglycemia. PMID- 19564456 TI - Microalbuminuria and cardiovascular autonomic dysfunction are independently associated with cardiovascular mortality: evidence for distinct pathways: the Hoorn Study. AB - OBJECTIVE: Microalbuminuria is associated with cardiovascular mortality, particularly among individuals with type 2 diabetes, but the mechanisms underlying this association are not completely understood. Microalbuminuria is known to be associated with cardiovascular autonomic dysfunction (C-AD), and C-AD in turn is associated with cardiovascular mortality. The purpose of this study, therefore, was to investigate whether C-AD can explain the relationship between microalbuminuria and cardiovascular mortality. RESEARCH DESIGN AND METHODS: We studied 490 individuals from a population-based cohort of individuals aged 50-75 years who were followed for a median period of 13.6 years. Microalbuminuria was defined as an albumin-to-creatinine ratio > or =2.0 mg/mmol in an early-morning spot-urine sample. Ten parameters reflecting different aspects of cardiovascular autonomic function were measured and compiled into a total score of C-AD (mean of separate z scores). The association between C-AD and microalbuminuria was estimated by multiple linear regression, and relative risks (RRs) for cardiovascular mortality were estimated by Cox proportional hazards analyses. RESULTS: After adjustments for age, sex, glucose tolerance status, and other risk factors, C-AD was associated with microalbuminuria (beta = 0.16 [95% CI 0.01 0.33]), and both microalbuminuria (RR 2.09 [1.07-4.08]) and C-AD (1.74 [1.04 2.89]) were associated with cardiovascular mortality. These associations did not change after further mutual adjustment for C-AD (2.13 [1.09-4.17]) or microalbuminuria (1.76 [1.05-2.94]), respectively. CONCLUSIONS: Both microalbuminuria and C-AD are independently associated with cardiovascular mortality, and the excess mortality attributable to microalbuminuria cannot be explained by C-AD. PMID- 19564455 TI - Endothelial markers may link kidney function to cardiovascular events in type 2 diabetes. AB - OBJECTIVE: The increased cardiovascular risk in diabetes has been linked to endothelial and renal dysfunction. The aim of this study was to investigate the role of stable fragments of the precursors of adrenomedullin, endothelin-1, vasopressin, and atrial natriuretic peptide in progression of cardiovascular disease in patients with diabetes. RESEARCH DESIGN AND METHODS: This was a prospective, observational study design with a composite end point (death or unexpected admission to hospital due to a cardiovascular event) on 781 patients with type 2 diabetes (54 events, median duration of observation 15 months). The four stable precursor peptides midregional adrenomedullin (MR-proADM), midregional proatrial natriuretic peptide (MR-proANP), COOH-terminal proendothelin-1 (CT-proET-1), and COOH-terminal provasopressin or copeptin (CT proAVP) were determined at baseline, and their association to renal function and cardiovascular events was studied using stepwise linear and Cox logistic regression analysis and receiver operating characteristic analysis, respectively. RESULTS: MR-proADM, CT-proET-1, CT-proAVP, and MR-proANP were all elevated in patients with future cardiovascular events and independently correlated to serum creatinine. MR-proADM and MR-proANP were significant predictors of a future cardiovascular event, with MR-proANP being the stronger (area under the curve 0.802 +/- 0.034, sensitivity 0.833, specificity 0.576, positive predictive value 0.132, and negative predictive value 0.978 with a cutoff value of 75 pmol/l). CONCLUSIONS: The four serum markers of vasoactive and natriuretic peptides are related to both kidney function and cardiovascular events, thus linking two major complications of diabetes, diabetic nephropathy and cardiovascular disease. PMID- 19564457 TI - Mortality and glycemic targets in the intensive care unit: another paradigm shift? PMID- 19564458 TI - Abnormal angiogenesis in diabetic nephropathy. PMID- 19564459 TI - Targeting B-cells mitigates autoimmune diabetes in NOD mice: what is plan B? PMID- 19564460 TI - Brown fat in humans: turning up the heat on obesity. PMID- 19564461 TI - (Pro)renin receptor: a treatment target for diabetic retinopathy? PMID- 19564462 TI - Bone disease, gestational diabetes mellitus, and health care. PMID- 19564463 TI - Pioglitazone-induced acute rhabdomyolysis. PMID- 19564464 TI - Ability of lipid accumulation product to identify metabolic syndrome in healthy men from Buenos Aires. PMID- 19564465 TI - Oral disposition index predicts the development of future diabetes above and beyond fasting and 2-h glucose levels: response to Utzschneider et al. PMID- 19564467 TI - Novel noninvasive breath test method for screening individuals at risk for diabetes: response to Dillon et al. PMID- 19564469 TI - Intensive glycemic control and the prevention of cardiovascular events: implications of the ACCORD, ADVANCE, and VA diabetes trials: a position statement of the American Diabetes Association and a scientific statement of the American College of Cardiology Foundation and the American Heart Association: response to Skyler et al. PMID- 19564471 TI - Hypoglycemia and clinical outcomes in patients with diabetes hospitalized in the general ward. AB - OBJECTIVE: Hypoglycemia is associated with adverse outcomes in mixed populations of patients in intensive care units. It is not known whether the same risks exist for diabetic patients who are less severely ill. In this study, we aimed to determine whether hypoglycemic episodes are associated with higher mortality in diabetic patients hospitalized in the general ward. RESEARCH DESIGN AND METHODS: This retrospective cohort study analyzed 4,368 admissions of 2,582 patients with diabetes hospitalized in the general ward of a teaching hospital between January 2003 and August 2004. The associations between the number and severity of hypoglycemic (6 months before the index pregnancy were included. Results were collected electronically via the DIAMOND Diabetes Information System. Pregnancy outcome was compared with background rates. RESULTS: There were 104 singleton pregnancies. The stillbirth rate (25/1,000) was 5 times, perinatal mortality rate (25/1,000) 3.5 times, and congenital malformation rate (24/1,000) 2 times that of the background population. A total of 28% of women received prepregnancy care, 43% received prepregnancy folic acid, and 51% achieved an A1C or=39 years, with BMI >or=25 kg/m(2) at baseline were selected. Baseline fasting serum samples from 160 individuals who developed type 2 diabetes and from 472 who did not were tested. An ultrasensitive immunoassay was used to measure of 58 candidate biomarkers in multiple diabetes associated pathways, along with six routine clinical variables. Statistical learning methods and permutation testing were used to select the most informative biomarkers. Risk model performance was estimated using a validated bootstrap bias correction procedure. RESULTS: A model using six biomarkers (adiponectin, C reactive protein, ferritin, interleukin-2 receptor A, glucose, and insulin) was developed for assessing an individual's 5-year risk of developing type 2 diabetes. This model has a bootstrap-estimated area under the curve of 0.76, which is greater than that for A1C, fasting plasma glucose, fasting serum insulin, BMI, sex-adjusted waist circumference, a model using fasting glucose and insulin, and a noninvasive clinical model. CONCLUSIONS: A model incorporating six circulating biomarkers provides an objective and quantitative estimate of the 5 year risk of developing type 2 diabetes, performs better than single risk indicators and a noninvasive clinical model, and provides better stratification than fasting plasma glucose alone. PMID- 19564474 TI - Effect of ingested interferon-alpha on beta-cell function in children with new onset type 1 diabetes. AB - OBJECTIVE: To evaluate the safety and efficacy of ingested human recombinant interferon-alpha (hrIFN-alpha) for preservation of beta-cell function in young patients with recent-onset type 1 diabetes. RESEARCH DESIGN AND METHODS: Subjects aged 3-25 years in whom type 1 diabetes was diagnosed within 6 weeks of enrollment were randomly assigned to receive ingested hrIFN-alpha at 5,000 or 30,000 units or placebo once daily for 1 year. The primary outcome was change in C-peptide secretion after a mixed meal. RESULTS: Individuals in the placebo group (n = 30) lost 56 +/- 29% of their C-peptide secretion from 0 to 12 months, expressed as area under the curve (AUC) in response to a mixed meal. In contrast, children treated with hrIFN-alpha lost 29 +/- 54 and 48 +/- 35% (for 5,000 [n = 27] and 30,000 units [n = 31], respectively, P = 0.028, ANOVA adjusted for age, baseline C-peptide AUC, and study site). Bonferroni post hoc analyses for placebo versus 5,000 units and placebo versus 30,000 units demonstrated that the overall trend was determined by the 5,000-unit treatment group. Adverse events occurred at similar rates in all treatment groups. CONCLUSIONS: Ingested hrIFN-alpha was safe at the doses used. Patients in the 5,000-unit hrIFN-alpha treatment group maintained more beta-cell function 1 year after study enrollment than individuals in the placebo group, whereas this effect was not observed in patients who received 30,000 units hrIFN-alpha. Further studies of low-dose ingested hrIFN alpha in new-onset type 1 diabetes are needed to confirm this effect. PMID- 19564475 TI - Anemia, diabetes, and chronic kidney disease. PMID- 19564477 TI - Redefining the diagnosis of diabetes using glycated hemoglobin. PMID- 19564476 TI - Hyperglycemic crises in adult patients with diabetes. PMID- 19564478 TI - Multiple biomarker prediction of type 2 diabetes. PMID- 19564479 TI - Point: Universal screening for gestational diabetes mellitus. PMID- 19564480 TI - Counterpoint: Selective screening for gestational diabetes mellitus. PMID- 19564481 TI - Public health implications of epigenetics. PMID- 19564483 TI - Recombinase-mediated cassette exchange provides a versatile platform for gene targeting: knockout of miR-31b. AB - A series of vectors has been designed to enhance the versatility of targeted homologous recombination. Recombinase-mediated cassette exchange permits sequential targeting at any locus and improves flexibility in making user-defined mutations. Application of RMCE to delete an intronic microRNA gene is described. PMID- 19564482 TI - Systems-level engineering of nonfermentative metabolism in yeast. AB - We designed and experimentally validated an in silico gene deletion strategy for engineering endogenous one-carbon (C1) metabolism in yeast. We used constraint based metabolic modeling and computer-aided gene knockout simulations to identify five genes (ALT2, FDH1, FDH2, FUM1, and ZWF1), which, when deleted in combination, predicted formic acid secretion in Saccharomyces cerevisiae under aerobic growth conditions. Once constructed, the quintuple mutant strain showed the predicted increase in formic acid secretion relative to a formate dehydrogenase mutant (fdh1 fdh2), while formic acid secretion in wild-type yeast was undetectable. Gene expression and physiological data generated post hoc identified a retrograde response to mitochondrial deficiency, which was confirmed by showing Rtg1-dependent NADH accumulation in the engineered yeast strain. Formal pathway analysis combined with gene expression data suggested specific modes of regulation that govern C1 metabolic flux in yeast. Specifically, we identified coordinated transcriptional regulation of C1 pathway enzymes and a positive flux control coefficient for the branch point enzyme 3-phosphoglycerate dehydrogenase (PGDH). Together, these results demonstrate that constraint-based models can identify seemingly unrelated mutations, which interact at a systems level across subcellular compartments to modulate flux through nonfermentative metabolic pathways. PMID- 19564484 TI - Alternative splicing of PTC7 in Saccharomyces cerevisiae determines protein localization. AB - It is well established that higher eukaryotes use alternative splicing to increase proteome complexity. In contrast, Saccharomyces cerevisiae, a single cell eukaryote, conducts predominantly regulated splicing through retention of nonfunctional introns. In this article we describe our discovery of a functional intron in the PTC7 (YHR076W) gene that can be alternatively spliced to create two mRNAs that code for distinct proteins. These two proteins localize to different cellular compartments and have distinct cellular roles. The protein translated from the spliced mRNA localizes to the mitochondria and its expression is carbon source dependent. In comparison, the protein translated from the unspliced mRNA contains a transmembrane domain, localizes to the nuclear envelope, and mediates the toxic effects of Latrunculin A exposure. In conclusion, we identified a definitive example of functional alternative splicing in S. cerevisiae that confers a measurable fitness benefit. PMID- 19564485 TI - A molecular mechanism of temperature sensitivity for mutations affecting the Drosophila muscle regulator Myocyte enhancer factor-2. AB - Temperature-sensitive (TS) mutations are a useful tool for elucidating gene function where a gene of interest is essential at multiple stages of development. However, the molecular mechanisms behind TS alleles vary. TS mutations of the myogenic regulator Myocyte enhancer factor-2 (MEF2) in Drosophila arise in the heteroallelic combination Mef2(30-5)/Mef2(44-5). We show that the 30-5 mutation affects the N-terminal MADS domain, causing impaired DNA binding ability and failure of homozygous mutants to survive to adulthood. The 44-5 mutation deletes a downstream splice acceptor site and results in a truncated protein that is unable to activate MEF2 targets. 44-5 homozygotes consequently show severely impaired myogenesis and die as embryos. We propose that in heteroallelic mutants at the permissive temperature, 30-5/44-5 heterodimers form and have a sufficiently stable interaction with DNA to activate myogenic gene expression; at the restrictive temperature, 44-5 homodimers displace 30-5/44-5 heterodimers from target genes, thus acting in a dominant-negative manner. To test this, we show that 30-5/44-5 heterodimers can form, and we study additional Mef2 alleles for complementation with the 30-5 allele. An allele affecting the DNA binding domain fails to complement 30-5, whereas two alleles affecting downstream residues show temperature-dependent complementation. Thus, by combining one MEF2 isoform having weakened DNA binding ability with a second truncated MEF2 mutant that has lost its activation ability, a TS form of intragenic complementation can be generated. These findings will provide new insight and guidance into the functions of dimeric proteins and how they might be engineered to generate TS combinations. PMID- 19564486 TI - Placental and embryonic growth restriction in mice with reduced function epidermal growth factor receptor alleles. AB - Embryos lacking an epidermal growth factor receptor (EGFR) exhibit strain specific defects in placental development that can result in mid-gestational embryonic lethality. To determine the level of EGFR signaling required for normal placental development, we characterized congenic strains homozygous for the hypomorphic Egfr(wa2) allele or heterozygous for the antimorphic Egfr(Wa5) allele. Egfr(wa2) homozygous embryos and placentas exhibit strain-dependent growth restriction at 15.5 days post-coitus while Egfr(Wa5) heterozygous placentas are only slightly reduced in size with no effect on embryonic growth. Egfr(wa2) homozygous placentas have a reduced spongiotrophoblast layer in some strains, while spongiotrophoblasts and glycogen cells are almost completely absent in others. Our results demonstrate that more EGFR signaling occurs in Egfr(Wa5) heterozygotes than in Egfr(wa2) homozygotes and suggest that Egfr(wa2) homozygous embryos model EGFR-mediated intrauterine growth restriction in humans. We also consistently observed differences between strains in wild-type placenta and embryo size as well as in the cellular composition and expression of trophoblast cell subtype markers and propose that differential expression in the placenta of Glut3, a glucose transporter essential for normal embryonic growth, may contribute to strain-dependent differences in intrauterine growth restriction caused by reduced EGFR activity. PMID- 19564488 TI - Histopathologic criteria to confirm white-nose syndrome in bats. AB - White-nose syndrome (WNS) is a cutaneous fungal disease of hibernating bats associated with a novel Geomyces sp. fungus. Currently, confirmation of WNS requires histopathologic examination. Invasion of living tissue distinguishes this fungal infection from those caused by conventional transmissible dermatophytes. Although fungal hyphae penetrate the connective tissue of glabrous skin and muzzle, there is typically no cellular inflammatory response in hibernating bats. Preferred tissue samples to diagnose this fungal infection are rostral muzzle with nose and wing membrane fixed in 10% neutral buffered formalin. To optimize detection, the muzzle is trimmed longitudinally, the wing membrane is rolled, and multiple cross-sections are embedded to increase the surface area examined. Periodic acid-Schiff stain is essential to discriminate the nonpigmented fungal hyphae and conidia. Fungal hyphae form cup-like epidermal erosions and ulcers in the wing membrane and pinna with involvement of underlying connective tissue. In addition, fungal hyphae are present in hair follicles and in sebaceous and apocrine glands of the muzzle with invasion of tissue surrounding adnexa. Fungal hyphae in tissues are branching and septate, but the diameter and shape of the hyphae may vary from parallel walls measuring 2 microm in diameter to irregular walls measuring 3-5 microm in diameter. When present on short aerial hyphae, curved conidia are approximately 2.5 microm wide and 7.5 microm in curved length. Conidia have a more deeply basophilic center, and one or both ends are usually blunt. Although WNS is a disease of hibernating bats, severe wing damage due to fungal hyphae may be seen in bats that have recently emerged from hibernation. These recently emerged bats also have a robust suppurative inflammatory response. PMID- 19564487 TI - Behavioral effects of gamma-hydroxybutyrate, its precursor gamma-butyrolactone, and GABA(B) receptor agonists: time course and differential antagonism by the GABA(B) receptor antagonist 3-aminopropyl(diethoxymethyl)phosphinic acid (CGP35348). AB - Gamma-hydroxybutyrate (GHB) is used therapeutically and recreationally. The mechanism by which GHB produces its therapeutic and recreational effects is not entirely clear, although GABA(B) receptors seem to play an important role. This role could be complex, because there are indications that different GABA(B) receptor mechanisms mediate the effects of GHB and the prototypical GABA(B) receptor agonist baclofen. To further explore possible differences in underlying GABA(B) receptor mechanisms, the present study examined the effects of GHB and baclofen on operant responding and their antagonism by the GABA(B) receptor antagonist 3-aminopropyl(diethoxymethyl)phosphinic acid (CGP35348). Pigeons were trained to peck a key for access to food during response periods that started at different times after the beginning of the session. In these pigeons, GHB, its precursor gamma-butyrolactone (GBL), and the GABA(B) receptor agonists baclofen and 3-aminopropyl(methyl)phosphinic acid hydrochloride (SKF97541) decreased the rate of responding in a dose- and time-dependent manner. CGP35348 shifted the dose-response curve of each agonist to the right, but the magnitude of the shift differed among the agonists. Schild analysis yielded a pA(2) value of CGP35348 to antagonize GHB and GBL [i.e., 3.9 (3.7-4.2)] that was different (P = 0.0011) from the pA(2) value to antagonize baclofen and SKF97541 [i.e., 4.5 (4.4-4.7)]. This finding is further evidence that the GABA(B) receptor mechanisms mediating the effects of GHB and prototypical GABA(B) receptor agonists are not identical. A better understanding of the similarities and differences between these mechanisms, and their involvement in the therapeutic effects of GHB and baclofen, could lead to more effective medications with fewer adverse effects. PMID- 19564489 TI - Systemic adenovirus infection in Sulawesi tortoises (Indotestudo forsteni) caused by a novel siadenovirus. AB - A novel siadenovirus was identified in the Sulawesi tortoise (Indotestudo forsteni). A group of 105 Sulawesi tortoises was obtained by the Turtle Survival Alliance. Many of the tortoises were in poor health. Clinical signs included anorexia, lethargy, mucosal ulcerations and palatine erosions of the oral cavity, nasal and ocular discharge, and diarrhea. Initial diagnostic tests included fecal testing for parasites, complete blood count and plasma biochemical analysis, mycoplasma serology, and polymerase chain reaction (PCR) testing for intranuclear coccidia and chelonian herpesvirus. Treatment included administration of antibiotics, antiparasitic medications, parenteral fluids, and nutritional support. Tissue samples from animals that died were submitted for histopathologic evaluation. Histopathologic examination revealed systemic inflammation and necrosis associated with intranuclear inclusions consistent with a systemic viral infection in 35 tortoises out of 50 examined. Fecal testing results and histopathologic findings revealed intestinal and hepatic amoebiasis and nematodiasis in 31 animals. Two of 5 tortoises tested by PCR were positive for Chlamydophila sp. Aeromonas hydrophila and Escherichia coli were cultured from multiple organs of 2 animals. The mycoplasma serology and PCR results for intranuclear coccidia and chelonian herpesvirus were negative. Polymerase chain reaction testing of tissues, plasma, and choanal/cloacal samples from 41 out of 42 tortoises tested were positive for an adenovirus, which was characterized by sequence analysis and molecular phylogenetic inference as a novel adenovirus of the genus Siadenovirus. The present report details the clinical and anatomic pathologic findings associated with systemic infection of Sulawesi tortoises by this novel Siadenovirus, which extends the known reptilian adenoviruses to the chelonians and extends the known genera of reptilian Adenoviridae beyond Atadenovirus to include the genus Siadenovirus. PMID- 19564490 TI - Diagnosis of intramammary infections at dry-off based on sampling strategy, epidemiology of pathogens, and agreement beyond chance. AB - Isolation of pathogens from duplicate or multiple milk samples is currently considered the gold standard in diagnosis of bovine intramammary infections (IMI). However, in large field studies and especially in normal dairy production conditions, collection of single samples is often the most practical option to determine the causal agents of mastitis in a herd. The objective of the present study was to determine how well results between the first and the second sample in pairs of duplicate and successive quarter milk samples agree, using 5 different IMI definitions, based on the number of colony forming units (CFU) per milliliter of milk and epidemiology of the pathogens isolated. Agreement between microbiologic results from the first and the second sample of a pair was assessed by calculating the percentage of agreement and kappa coefficient. Milk samples collected at dry-off from 561 Holstein cows in 4 Ohio dairy herds were included in the analyses. Results of the study indicate that the agreement between the first and the second sample of a duplicate pair was high when criteria to call a sample positive was adjusted for the number of CFU/ml of milk by considering the epidemiology of different mastitis organisms. This finding suggests that an IMI can be accurately diagnosed with single samples. For contagious pathogens (Staphylococcus aureus) a cutoff of 100 CFU/ml and a cutoff of 1,000 CFU/ml for major environmental and minor pathogens will serve as a sensible approach to diagnose bovine IMI with single milk samples. PMID- 19564491 TI - Infectious and lethal doses of H5N1 highly pathogenic avian influenza virus for house sparrows (Passer domesticus) and rock pigeons (Columbia livia). AB - Terrestrial wild birds commonly associated with poultry farms have the potential to contribute to the spread of H5N1 highly pathogenic avian influenza (HPAI) virus within or between poultry facilities or between domesticated and wild bird populations. This potential, however, varies between species and is dependent on several virus and host factors, including habitat utilization, susceptibility, and viral shedding patterns. To provide data on susceptibility and shedding patterns of house sparrows (Passer domesticus) and rock pigeons (Columba livia), 20 birds from each species were inoculated with decreasing concentrations of A/whooper swan/Mongolia/244/05 (H5N1) HPAI virus, and the birds were evaluated for morbidity, mortality, viral shedding, and seroconversion over a 14-day trial. The house sparrows were highly susceptible to the H5N1 HPAI virus as evidenced by low infectious and lethal viral doses. In addition, house sparrows excreted virus via the oropharynx and cloaca for several days prior to the onset of clinical signs. Based on these results, house sparrows could play a role in the dissemination of H5N1 HPAI virus in poultry. In contrast, pigeons were resistant to the HPAI virus, requiring a high concentration of virus to produce infection or death. When infection did occur, the duration of viral shedding was brief, and viral titers were low. The data suggests that pigeons would contribute little to the transmission and spread of H5N1 HPAI virus in poultry. PMID- 19564492 TI - Bayesian estimation of the performance of using clinical observations and harvest lung lesions for diagnosing bovine respiratory disease in post-weaned beef calves. AB - Bovine respiratory disease (BRD) diagnosis during the postweaning phase of beef production is an important component of effective preventive health and treatment programs. Although identification of diseased animals based on signs of clinical illness (CI) is a common method in the beef industry for identifying BRD, very little information is available on the accuracy of this method. Previous investigators hypothesized that monitoring pulmonary lesions at harvest (LU) could be a more reliable indicator of disease status during the postweaning phase. A structured literature review was conducted to identify research that compared CI and LU. Because there is no true gold standard for diagnosing BRD, Bayesian methods were used to estimate the sensitivity and specificity of each diagnostic method relative to a BRD diagnosis at any time during the postweaning phase. Results from the current study indicate that the estimated diagnostic sensitivity and specificity of CI were 61.8% (97.5% probability interval [PI]: 55.7, 68.4) and 62.8% (97.5% PI: 60.0, 65.7), respectively. Use of LU for a BRD diagnosis was estimated to have a sensitivity of 77.4% (97.5% PI: 66.2, 87.3) and a specificity of 89.7% (97.5% PI: 86.0, 93.8). Further analysis revealed that the probabilities of LU having higher sensitivity and specificity than CI were 99.4% and 100%, respectively. The present research indicates that neither method was perfect, and both methods were relatively poor at correctly classifying truly diseased animals (sensitivity) but that LU was more accurate than CI for BRD diagnosis. Results from the present study should be considered when these diagnostic methods are used to evaluate BRD outcomes in clinical and research settings. PMID- 19564493 TI - Nor98 scrapie identified in the United States. AB - A distinct strain of scrapie identified in sheep of Norway in 1998 has since been identified in numerous countries throughout Europe. The disease is known as Nor98 or Nor98-like scrapie, among other names. Distinctions between classic scrapie and Nor98 scrapie are made based on histopathology and immunodiagnostic results. There are also differences in the epidemiology, typical signalment, and likelihood of clinical signs being observed. In addition, sheep that have genotypes associated with resistance to classic scrapie are not spared from Nor98 disease. The various differences between classic and Nor98 scrapie have been consistently reported in the vast majority of cases described across Europe. The current study describes in detail the pathologic changes and diagnostic results of the first 6 cases of Nor98 scrapie disease diagnosed in sheep of the United States. PMID- 19564494 TI - Lung pathology and infectious agents in fatal feedlot pneumonias and relationship with mortality, disease onset, and treatments. AB - This study charted 237 fatal cases of bovine respiratory disease (BRD) observed from May 2002 to May 2003 in a single Oklahoma feed yard. Postmortem lung samples were used for agent identification and histopathology. Late in the study, 94 skin samples (ear notches) were tested for Bovine viral diarrhea virus (BVDV) by immunohistochemistry (IHC). Bovine respiratory disease morbidity was 14.7%, and the mortality rate of all causes was 1.3%, with more than half (53.8%) attributed to BRD (0.7% total of all causes). The agents isolated were the following: Mannheimia haemolytica (25.0%), Pasteurella multocida (24.5%), Histophilus somni (10.0%), Arcanobacterium pyogenes (35.0%), Salmonella spp. (0.5%), and Mycoplasma spp. (71.4%). Viruses recovered by cell culture were BVDV-1a noncytopathic (NCP; 2.7%), BVDV-1a cytopathic (CP) vaccine strain (1.8%), BVDV-1b NCP (2.7%), BVDV-2a NCP (3.2%), BVDV-2b CP (0.5%), and Bovine herpesvirus 1 (2.3%). Gel-based polymerase chain reaction (PCR) assays were 4.6% positive for Bovine respiratory syncytial virus and 10.8% positive for Bovine coronavirus. Bovine viral diarrhea virus IHC testing was positive in 5.3% of the animals. The mean values were determined for the treatment data: fatal disease onset (32.65 days), treatment interval (29.15 days), number of antibiotic treatments (2.65), number of different antibiotics (1.89), and day of death (61.81 days). Lesions included the following: 1) duration: acute (21%), subacute (15%), chronic (40.2%), healing (2.8%), normal (18.1%), and autolyzed (2.8%); 2) type of pneumonia: lobar bronchopneumonia (LBP; 27.1%), LBP with pleuritis (49.1%), interstitial pneumonia (5.1%), bronchointerstitial pneumonia (1.4%), septic (0.9%), embolic foci (0.5%), other (2.8%), normal (10.3%), and autolyzed (2.8%); and 3) bronchiolar lesions: bronchiolitis obliterans (39.7%), bronchiolar necrosis (26.6%), bronchiolitis obliterans/bronchiolar necrosis (1.4%), other bronchiolar lesions (6.5%), and bronchiolar lesion negative (25.7%). Statistically significant relationships were present among the agents, lesions, and the animal treatment, disease onset, and mortality data. Clinical illnesses observed in this study were lengthier than those reported 16-20 years ago, based on fatal disease onset, treatment interval, and day of death. PMID- 19564495 TI - Axonal degeneration in sheep caused by the ingestion of Halimium brasiliense. AB - Nervous system disease is reported in sheep from 2 farms in southern Brazil and in 33 farms in Uruguay. The illness was seasonal, occurring from May to November, during the growing season of Halimium brasiliense, and primarily affected sheep older than 3 years of age. Clinical signs included transient seizures that occurred mainly when sheep were disturbed or frightened. Most affected sheep recovered when removed to other pastures. Feeding trials produced clinical signs in 1 sheep after the ingestion of 2,117 g/kg of body weight of H. brasiliense over 142 days. Two sheep that had previously recovered from spontaneous toxicosis developed clinical signs after the ingestion of 263 g and 565 g of H. brasiliense per kg body weight given over 36 and 31 days, respectively. The main histologic lesion was vacuolation of the brain and spinal cord, with rare axonal spheroid formation. Transmission electron microscopy revealed segmental axonal swelling with degeneration and disappearance of the axonal organelles and vacuolation of the axoplasm. A pigment identified as ceroid was also present in neurons, astrocytes, and macrophages. These lesions suggested a novel morphologic manifestation of a toxic axonopathy. PMID- 19564496 TI - Characterization of Cervidpoxvirus isolates from Oregon, California, and eastern Canada. AB - The present report describes the analysis of 4 Deerpox virus isolates from California, Oregon, and Ontario, Canada. All 4 isolates were associated with cutaneous crusting lesions. Examination of selected samples by electron microscopy demonstrated that the viruses were morphologically similar to orthopoxviruses. Phylogenetic analysis of the A21 gene, which is found in all poxviruses, indicated that the 4 isolates form a lineage distinct from other members except for those belonging to the genus Cervidpoxvirus of the subfamily Chordopoxvirinae. Members of the Cervidpoxvirus lineage encode a set of genes not found in other poxviruses. These include homologs of genes encoding interleukin 1 receptor antagonists (IL-1Ra) and C-type lectin-like receptors (CTLR). In the current investigation, genes encoding homologs of IL-1Ra and CTLR were amplified from all the isolates and were found to be closely related to orthologs found in the Cervidpoxvirus genus, which further supports the inclusion of these isolates in the Cervidpoxvirus genus. PMID- 19564497 TI - Genetic variability of archived Cytauxzoon felis histologic specimens from domestic cats in Georgia, 1995-2007. AB - Cytauxzoon felis infection in domestic cats has historically been nearly 100% fatal. However, increasing reports of domestic cats that survive cytauxzoonosis and reports of asymptomatic cats with C. felis infections suggest the existence of different parasite strains that vary in pathogenicity. The objective of the current study was to obtain epidemiologic information about cytauxzoonosis through genotypic characterization of archived histologic specimens from domestic cats with C. felis infections that were diagnosed in Georgia between 1995 and 2007. Such retrospective data on genetic variability will provide an historic context for current studies of C. felis genotype frequencies. Cytauxzoon felis DNA was obtained from formalin-fixed, paraffin-embedded tissues from infected cats diagnosed with cytauxzoonosis at necropsy. Genetic characterization of C. felis was performed using sequence analysis of the polymerase chain reaction amplified ribosomal internal transcribed spacer regions 1 and 2 (ITS1, ITS2). Eleven different combined ITS1 and ITS2 sequences were identified, the majority of which were identical to those previously reported in fatally infected cats from Georgia. The findings of the current study document the existence of genetically distinct C. felis populations in historical samples and, together with data from contemporary samples, demonstrate a diverse population structure for C. felis. PMID- 19564498 TI - Neutralizing monoclonal antibody sandwich liquid-phase blocking enzyme-linked immunosorbent assay for detection of Foot-and-mouth disease virus type O antibodies. AB - Liquid-phase blocking enzyme-linked immunosorbent assay (LPBE) using the neutralizing monoclonal antibody (mAb) sandwich method (M-LPBE) for detection of Foot-and-mouth disease virus (FMDV) type O antibodies was developed. Two neutralizing mAbs, 72C1 and 65H6, were raised against the FMDV O/JPN/2000 strain, and used as trapping and peroxidase-labeled detecting antibodies, respectively. Sera from animals experimentally infected with FMDV showed specific positive results by M-LPBE, which were correlated with the results of the virus neutralization test (VNT). When 303 negative bovine and 302 negative swine sera were tested, the specificity of M-LPBE was 100% and 99.7%, respectively. In addition, nine samples that had been collected in 2000 in Japan and regarded as evidently false positives by LPBE (supplied by the World Reference Laboratory for Foot-and-Mouth Disease) were uniformly negative by M-LPBE, just like VNT. Therefore, M-LPBE seems to have sufficient specificity for FMDV type O antibody screening and diagnosis. PMID- 19564499 TI - Identification of major antigenic proteins of Edwardsiella tarda recognized by Japanese flounder antibody. AB - Edwardsiella tarda is a fish pathogen that causes systemic infections in fresh water and marine fish. Determining the antigenic proteins is important for the development of an immunodiagnostic tests and a vaccine for effective infection control in fish. In the current study, antigens were detected by immunoblotting and affinity column chromatography using a Japanese flounder (Paralichthys olivaceus) antibody produced by experimental infection with E. tarda. GroEL, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), outer membrane protein A, filament protein, 30S ribosomal protein S6, 50S ribosomal protein L9, cold shock protein, and carbon storage protein were identified as antigens of E. tarda through biochemical analyses of the molecular weights, isoelectric points, and N terminal amino-acid sequences. These proteins can be easily detected in flounder infected with E. tarda and are potential diagnostic markers. PMID- 19564500 TI - Genotypes, antibiogram, and pulsed-field gel electrophoresis profiles of Escherichia coli strains from piglets in Korea. AB - Adherence factors and enterotoxins are major virulence factors of enterotoxigenic Escherichia coli (ETEC). Antibiotics have been used frequently for the treatment and prevention of ETEC infection in piggeries worldwide, including Korea. Therefore, data on both virulence profiles and antibiotic resistance patterns are useful in the epidemiological study of ETEC. A total number of 198 E. coli field isolates were examined. The most prevalent pathotype was F1, followed by a combination of F1 and EAST1. All of the 71 isolates were resistant to more than 2 antibiotics used in a disk diffusion test, and 87.94% of the isolates were found to be resistant to more than 4 antibiotics. Investigations were also conducted to correlate the virulence gene profiles with antibiogram and pulsed-field gel electrophoresis (PFGE). Although a high degree of polymorphism was noted among strains having the same virulence patterns, the highest similarity pattern was observed carrying the same virulence profiles and similar antibiogram. Thus, investigation of both virulence profiles and antibiogram is essential to the epidemiological study of ETEC. Moreover, the PFGE method might be applicable as a tool to reveal genetic relatedness among E. coli strains from piggeries in Korea. PMID- 19564501 TI - Rapid bilateral intraocular cocktail sampling method for West Nile virus detection in dead corvids. AB - Corvids can be a sensitive indicator for West Nile virus (WNV) prevalence and are a component of many WNV surveillance programs. An improved sampling procedure using a bilateral intraocular cocktail (BIC) was developed for testing corvid carcasses for WNV. This new procedure was substantially faster than harvesting internal organs, required less specialized equipment and training, and yielded excellent diagnostic sensitivity. PMID- 19564502 TI - Detection of Avian leukosis virus genome by a nested polymerase chain reaction using DNA and RNA from dried feather shafts. AB - The nested polymerase chain reaction (nPCR) using frozen feather pulp is useful for detecting fowl glioma-inducing virus (FGV), which belongs to the Avian leukosis virus family, and it has recently been suggested that FGV has spread to ornamental chickens kept in Japanese zoological gardens. In the current study, the practicality of using DNA and RNA from dried feather shafts as PCR samples was examined to establish a simple method for tissue preservation. Feather shafts were collected from 7 FGV-positive chickens and stored at room temperature for 30 days. DNA and RNA were extracted from these dried materials. All DNA and complementary DNA (cDNA) prepared from the RNA showed positive results for chicken beta-actin and FGV, respectively. Screening for FGV was performed on Japanese fowls kept in zoological garden N. Of the feather shafts collected from 57 birds, 1 sample tested positive for FGV according to PCR of DNA and cDNA samples from the dried feather shafts. This positive bird originated from zoological garden A and had brain lesions suggestive of fowl glioma. The results suggest that DNA and RNA from dried feather shafts can be used in nPCR to detect the FGV genome. PMID- 19564503 TI - Evaluation of a blocking enzyme-linked immunosorbent assay for serological diagnosis of Bovine herpesvirus 1. AB - Bovine herpesvirus 1 (BoHV-1) is an infectious agent of concern in the international export of bovine products; it is endemic in the United States, but it has been eradicated in many countries of the European Union (EU). For export of semen to the EU, accurate assessment of BoHV-1 status of the bull is required and is usually accomplished by measuring the level of antibody to the virus. The gold standard is virus neutralization (VN) using overnight incubation with the virus, a test approved by the World Organization for Animal Health (OIE). Enzyme linked immunosorbent assay (ELISA) is also approved for international trade. The lone U.S. Department of Agriculture-approved commercial ELISA was compromised with specificity problems, which necessitated the development of a different ELISA. Of 4 monoclonal antibodies evaluated, 1 directed against glycoprotein C of BoHV-1 was found to be the most reliable. One hundred twenty-eight characterized positive samples and 334 negative serum samples were tested. The blocking ELISA showed 97.7% sensitivity and 99.4% specificity as compared with OIE VN. The Wisconsin Veterinary Diagnostic Laboratory ELISA fulfills the OIE requirement for a blocking or competitive ELISA to qualify animals for export to BoHV-1-free countries. PMID- 19564504 TI - Naturally occurring influenza infection in a ferret (Mustela putorius furo) colony. AB - Tissue samples from 2 juvenile ferrets (Mustela putorius furo) from a colony that was undergoing an outbreak of respiratory disease were submitted to the Iowa State University Veterinary Diagnostic Laboratory. Microscopic examination of lung samples revealed bronchointerstitial pneumonia with necrotizing bronchiolitis. Influenza A virus was detected in sections of formalin-fixed lung by immunohistochemistry and reverse transcription polymerase chain reaction assay. A field investigation of the premises and analysis of additional samples led to the confirmation and characterization of an influenza virus with high homology to contemporary reassortant H1N1 swine influenza viruses. Although ferrets have been used extensively to research the virulence and transmissibility of avian, human, and swine influenza virus strains, no published information exists on naturally occurring outbreaks of swine influenza in ferrets. PMID- 19564505 TI - Herpesvirus infections in rock hyraxes (Procavia capensis). AB - Seven juveniles and 3 adults from a closed group of 19 rock hyraxes (Procavia capensis) housed in a zoo's indoor rock exhibit died or were euthanized after developing blepharoconjunctivitis and orofacial ulcers over a 2-week period. Histopathologic examination of dermal ulcers and ulcerated tongues revealed amphophilic to basophilic intranuclear inclusion bodies in epithelial cells bordering ulcers. Epithelial cells with inclusion bodies were often characterized by cytomegaly and karyomegaly, and many cells had formed syncytia. Examination of inclusion bodies in tongue epithelium by transmission electron microscopy revealed icosahedral nucleocapsids, approximately 80-95 nm in diameter, with morphologic features consistent with herpesvirus. Cytopathic effect (CPE) typical of alphaherpesvirus infection was seen in bovine turbinate, equine dermal, and Vero cell monolayers after inoculation with homogenates of the skin lesions, but CPE was not seen after inoculation onto Madin-Darby canine kidney or swine testicle cell monolayers. Polymerase chain reaction analysis using degenerate primers that targeted a portion of the herpesvirus polymerase gene generated a product of approximately 227 base pairs. The product was cloned, sequenced, and then analyzed using BLAST. At the nucleotide level, there was 86%, 77%, and 76% shared identity with Eidolon herpesvirus 1, Human herpesviruses 1 and 2, and Cercopithecine herpesvirus 2, respectively. Herpesvirus infections in rock hyraxes have not been characterized. The data presented in the current study suggest that a novel alphaherpesvirus caused the lesions seen in these rock hyraxes. The molecular characteristics of this virus would tentatively support its inclusion in the genus Simplexvirus. PMID- 19564506 TI - Clinical, histologic, and immunohistochemical features of an undifferentiated renal tubular carcinoma in a juvenile olive baboon (Papio anubis). AB - An undifferentiated renal tubular carcinoma was diagnosed in a juvenile male olive baboon (Papio anubis). The animal suddenly appeared depressed and refused to eat. During physical examination, a firm, palpable mass in the left abdominal area and flank pain were detected. Clinical pathology findings included mild anemia, hypoalbuminemia, hyponatremia, and mildly increased serum creatinine and urea concentrations. Radiographs revealed a large mass in the left abdominal area. Exploratory laparotomy disclosed a 10 cmx15 cm multilobulated mass involving the left kidney and adjacent organs. Because of a poor prognosis, the animal was humanely euthanized, and necropsy was performed. Tissue samples of the neoplasm were taken for histopathological examination. Immunohistochemical staining was done using vimentin, cytokeratin, S-100 protein, Ki-67, alpha-actin, and desmin-specific primary antibodies. Microscopically, elongated and irregular tubules were lined by 2 or more layers of atypical epithelial cells. Anisocytosis, anisokaryosis, and frequent mitotic figures were also observed. Following immunohistochemical staining, the cytoplasm of neoplastic cells was positive for cytokeratin, vimentin, and S-100 protein and negative for alpha actin and desmin. Positive nuclear staining for Ki-67 was observed. The neoplasm was diagnosed as an undifferentiated renal tubular carcinoma. PMID- 19564507 TI - Chronic copper toxicosis in sheep following the use of copper sulfate as a fungicide on fruit trees. AB - Between January and October 2006, 15 Chios sheep died in a field located near a factory in Orhangazi, Bursa, Turkey. In addition, in May 2007, 2 ewes died after aborting in the same field. Clinical signs in affected animals prior to death were anorexia, hematuria, icterus, incoordination, and ptyalism. Postmortem findings included generalized icterus; yellow, friable livers; distended gallbladders with dense, dark bile; and dark, hypertrophic kidneys with hemorrhage. Copper (Cu) concentrations were measured in multiple specimens of the following: 9 sera, 3 livers, 3 kidneys, 4 plants (including 2 artichoke leaf specimens), 3 soil samples, and 1 drinking water sample. High Cu concentrations were present in the livers, kidneys, and sera of dead sheep, as well as in the vegetation and soil samples from the field. Chronic Cu toxicosis was confirmed as the cause of death attributed primarily to the use of copper sulfate as a fungicide for fruit trees within the field. In addition, factory dust containing Cu might have been an additional factor in the toxicosis. PMID- 19564508 TI - Persistent truncus arteriosus with dissecting aneurysm and subsequent cardiac tamponade in a lamb. AB - The present case report describes the necropsy and histopathology findings of a 3 week-old lamb with persistent truncus arteriosus (PTA). This rare cardiac malformation was characterized by the presence of a common arterial trunk arising from the right ventricle and overriding a ventricular septal defect. The pulmonary arteries originated from a short common trunk from this PTA, which subsequently continued as the thoracic aorta. The death of the lamb was attributed to a rupture of the PTA with subsequent cardiac tamponade. Histologically, a dissecting aneurysm with elastic fiber fragmentation in the wall of the PTA was identified as the underlying pathologic condition. Altered hemodynamic forces with subsequent secondary vasculopathy, as well as congenital primary vasculopathies, should be considered as an underlying pathogenetic mechanism. PMID- 19564509 TI - Intraocular sarcomas in two rabbits. AB - Intraocular neoplasms are described in 2 adult rabbits. The left globe of an 8 year-old male rabbit was enucleated after chronic inflammatory disease resulted in a nonvisual eye. The left globe of a 5-year-old female rabbit also was enucleated after a history of lens-induced uveitis, cataract formation, and resultant glaucoma. In both rabbits, histopathology revealed a variably pleomorphic, poorly differentiated, invasive, intraocular spindle cell neoplasm closely associated with lens and lens capsular fragments. Gram stains failed to detect bacterial organisms or Encephalitozoon cuniculi. Polymerase chain reaction assays, used to amplify the 16S RNA gene of numerous bacteria and E. cuniculi, were also negative. Immunohistochemical staining demonstrated strong, diffuse expression for vimentin; however, staining for smooth muscle actin, cytokeratin, S100, and desmin were negative. Long-standing intraocular inflammation and/or traumatic insults to the eyes were considered as causes of these neoplasms. The histologic features of these intraocular neoplasms closely resemble post traumatic ocular sarcomas in cats. PMID- 19564510 TI - Cutaneous viral papilloma with local extension and subungual cyst formation in a dog. AB - A viral-induced digital cutaneous exophytic papilloma was diagnosed in a 2-year old, spayed, female Siberian husky dog with lameness. Digital pain and lameness persisted after removal of the initial papilloma, and the fifth lateral digit was subsequently amputated. Upon histologic examination of the digit, a de novo digital, cutaneous, inverted, viral papilloma and subungual cyst were diagnosed. The inverted cutaneous papilloma, located at the junction of the digital paw pad and ventral nail, extended focally through the nail into the subungual space, where an expansile cyst was formed. Cellular changes suggestive of papillomavirus infection were present in the epithelium of the original exophytic papilloma, as well as the endophytic mass and subungual cyst. Cytopathic effects included ballooning degeneration of keratinocytes, koilocytosis, irregularity of keratohyalin granules, and margination of nuclear chromatin. Numerous faintly basophilic to eosinophilic intranuclear inclusions measuring 10-15 microm in diameter were present within keratinocytes of the exophytic, endophytic, and subungual cystic lesions. Electron microscopy was performed on tissues from all lesions and revealed numerous 40-45 nm diameter hexagonal virions characteristic of papillomavirus that were arranged in crystalline arrays and dense clusters within affected nuclei. PMID- 19564511 TI - Mycotic pododermatitis and mycotic pneumonia in commercial turkey poults in northern California. AB - Seven 5-week-old broad-breasted white commercial meat turkeys were submitted to the California Animal Health and Food Safety laboratory in Turlock with a history of respiratory illness. The primary diagnostic findings were mycotic pododermatitis and mycotic pneumonia. The unique feature of this case was the colonization of footpad epidermis and subcutis by fungal hyphae in commercial turkey species. No fungal cultures were undertaken at the time of the necropsy; therefore, paraffin-embedded tissue sections of lung and footpads were used to extract, amplify, and sequence mycotic DNA. A mixed population of fungi was identified in both lung and footpads by polymerase chain reaction amplification of part of the large subunit ribosomal RNA gene using broad-range fungal primers and DNA sequencing. In footpads, sequences matching Cryptococcus saitoi and Cladosporium and Cudoniella species were identified. It is believed that these fungi were opportunistic pathogens originating from the litter. The fungi identified from lungs were Aspergillus species, most closely matching Aspergillus flavus and Arxiozyma telluris (most likely a contaminant). Mycotic pododermatitis in avian species is considered a rare pathologic finding, and few documented reports are available. The on-farm prevalence of footpad lesions was estimated at 3%, and there was no associated increase in the incidence of lameness or weight depression in affected birds. Microscopically, a granulomatous inflammatory reaction associated with fungal hyphae was observed in lung parenchyma. Disruption of keratinized epidermis, encrustations, and acute inflammation were also noted in footpads invaded with fungal hyphae. PMID- 19564512 TI - Nephrotoxicosis in Iberian piglets subsequent to exposure to melamine and derivatives in Spain between 2003 and 2006. AB - Between November 2003 and September 2006, 300 to 400 45-60-day-old Iberian piglets developed anorexia, polydipsia, and lethargy. Piglets were from 5 different farms in the western part of Spain. Morbidity was between 40% and 60%, and mortality ranged from 20% to 40% of the total population of postweaning piglets. In the 9 piglets in which postmortem examinations were conducted, kidneys were enlarged with yellow foci in the cortex and medulla. Microscopically, these foci were accumulations of crystals within the lumina of dilated distal tubules and collecting ducts, causing flattening of the renal tubular epithelial cells. The crystals displayed a multicolored birefringence under cross-polarized light. The multinucleated giant cells surrounding the crystals, interstitial fibrosis, and nonsuppurative infiltrates indicated a chronic inflammatory response. Toxicologic analysis of fixed kidney tissues from 4 piglets demonstrated the presence of melamine, ammeline, ammelide, and cyanuric acid. Ammelide concentrations were highest, ranging from 39,000 to 92,000 mg/kg, followed by ammeline (20,000-34,000 mg/kg), melamine (9,200-29,000 mg/kg), and cyanuric acid (2,200-9,100 mg/kg). The clinical, histologic, and toxicologic findings in affected piglets were similar to those reported in dogs and cats that died of melamine and melamine analogue-associated renal failure in 2004 and 2007. To the authors' knowledge, this is the first documented report of poisoning due to melamine and its analogues in pigs and demonstrates that contamination of pig feed occurred as early as 2003. PMID- 19564513 TI - Pleomorphic leiomyosarcoma in the hind leg of a Taiwanese macaque (Macaca cyclopis). AB - An adult, captive Taiwanese monkey (Macaca cyclopis) presented clinically with a large, rapidly growing mass located in the crus of the left hind leg. The overlying skin was severely ulcerated and necrotic. Radiographs suggested an invasive neoplasm in soft tissue with no bone involvement. The animal's clinical condition progressively worsened over the next 3 months until it died. Necropsy revealed that the mass infiltrated the surrounding skeletal muscle with no evidence of distant metastasis. Microscopically, the mass was highly cellular and composed of round to spindle cells with frequent rhabdoid cells characterized by abundant eosinophilic glassy cytoplasmic inclusions and large, bizarre nuclei. Phosphotungstic acid hematoxylin staining failed to reveal distinct cross striations within the neoplastic cells. Neoplastic cells were strongly positive for smooth muscle actin and vimentin but were negative for sarcomeric actin, myoglobin, desmin, cytokeratin, S100, and lysozyme. The gross, microscopic, and immunohistochemical findings supported the diagnosis of pleomorphic leiomyosarcoma. PMID- 19564514 TI - Rhabditid nematode-associated ophthalmitis and meningoencephalomyelitis in captive Asian horned frogs (Megophrys montana). AB - Between 2006 and 2008, 4 captive Asian horned frogs (Megophrys montana) were diagnosed with ocular and neurologic disease associated with rhabditid nematodiasis. Mortality, either spontaneous or by humane euthanasia, was high (3/4, 75%). Gross and histologic findings included varying degrees of ulcerative keratitis, histiocytic uveitis and retinitis, meningoencephalomyelitis, and epidermal chromatophore (iridophore) hyperplasia with intralesional nematodes. Entry into the host was presumed to be by direct invasion of the skin and the cornea with migration through the optic nerve to the brain and spinal cord. One frog was diagnosed with rhabditid nematodiasis antemortem, and clinical signs and lesions in the frog did not progress after unilateral enucleation and anthelminthic treatment were completed. Gross and tissue morphology of the nematodes were consistent with the order Rhabditida. DNA was extracted separately from 2 individual nematodes that were isolated from frozen and ethanol-preserved eye and brain tissue. These DNA templates were used for polymerase chain reaction amplification and sequencing of nuclear 28S large subunit (LSU) and internal transcribed spacer (ITS) ribosomal DNA regions. Comparison of the LSU and ITS sequences to those deposited in GenBank revealed an exact match for Caenorhabditis elegans. PMID- 19564516 TI - The ability of the quadruple test to predict adverse perinatal outcomes in a high risk obstetric population. AB - OBJECTIVE: To determine the ability of the quadruple Down's syndrome screening test (quad screen) to predict other adverse perinatal outcomes (APO) in a high risk obstetric population. SETTING: A tertiary medical centre in West Virginia. METHODS: We retrospectively reviewed 342 obstetric patients with quad screen data from a single clinic. The quad screen included maternal serum levels of alphafetoprotein (AFP), human chorionic gonadotrophin (hCG), uncongjugated oestriol (uE(3)), and inhibin A. The risk of APO was compared between patients with at least one abnormal marker versus no abnormal markers and >or=2 abnormal markers versus <2 abnormal markers. Abnormal markers were determined by cut-off values produced by Receiver Operator Characteristic (ROC) curves and the FASTER trial. Unadjusted and adjusted effects were estimated using logistic regression analysis. RESULTS: The risk of having an APO increased significantly for patients with abnormal markers by about three-fold using ROC and two-fold using FASTER trial thresholds. CONCLUSIONS: The quad screen shows value in predicting risk of APO in high-risk patients. PMID- 19564518 TI - League tables of breast cancer screening units: worst-case and best-case scenario ratings helped in exposing real differences between performance ratings. AB - OBJECTIVES: Data on the performance of health boards, hospitals and medical specialists, etc., are being collected at various levels in the health-care system and are often presented as league tables. These tables ignore natural variation and/or confounders, and this introduces uncertainty about their interpretation. The purpose of this study was to devise and illustrate a method to expose the real difference between the ratings in league tables. METHODS: Two values per rating were added to the league tables: the best-case scenario and the worst-case scenario. True performance will lie somewhere between these two values. The method is illustrated using data from the Dutch breast cancer screening programme. RESULTS: By focusing on one performance indicator and one confounder, it was possible to show shifts in the rating order of breast cancer screening units and thus expose the uncertainty about the true performance of each screening unit. CONCLUSIONS: The worst-case and best-case scenario ratings demonstrated the uncertainty within the ratings of a league table. League tables should therefore only be used with great caution and after providing the public with sufficient information. PMID- 19564517 TI - Demographic variations in HIV testing history among emergency department patients: implications for HIV screening in US emergency departments. AB - OBJECTIVE: To determine the proportion of emergency department (ED) patients who have been tested for human immunodeficiency virus (HIV) infection and assess if patient history of HIV testing varies according to patient demographic characteristics. DESIGN: From July 2005-July 2006, a random sample of 18-55-year old English-speaking patients being treated for sub-critical injury or illness at a northeastern US ED were interviewed on their history of HIV testing. Logistic regression models were created to compare patients by their history of being tested for HIV according to their demography. Odds ratios (ORs) with 95% confidence intervals (CIs) were estimated. RESULTS: Of 2107 patients surveyed who were not known to be HIV-infected, the median age was 32 years; 54% were male, 71% were white, and 45% were single/never married; 49% had private health-care insurance and 45% had never been tested for HIV. Of the 946 never previously tested for HIV, 56.1% did not consider themselves at risk for HIV. In multivariable logistic regression analyses, those less likely to have been HIV tested were male (OR: 1.32 [1.37-2.73]), white (OR: 1.93 [1.37-2.73]), married (OR: 1.53 [1.12-2.08]), and had private health-care insurance (OR: 2.10 [1.69 2.61]). There was a U-shaped relationship between age and history of being tested for HIV; younger and older patients were less likely to have been tested. History of HIV testing and years of formal education were not related. CONCLUSION: Almost half of ED patients surveyed had never been tested for HIV. Certain demographic groups are being missed though HIV diagnostic testing and screening programmes in other settings. These groups could potentially be reached through universal screening. PMID- 19564519 TI - Differences in endpoints between the Swedish W-E (two county) trial of mammographic screening and the Swedish overview: methodological consequences. AB - OBJECTIVES: To characterize and quantify the differences in the number of cases and breast cancer deaths in the Swedish W-E Trial compared with the Swedish Overview Committee (OVC) summaries and to study methodological issues related to trials in secondary prevention. SETTING: The study population of the W-E Trial of mammography screening was included in the first (W and E county) and the second (E-county) OVC summary of all Swedish randomized mammography screening trials. The OVC and the W-E Trial used different criteria for case definition and causes of death determination. METHOD: A Review Committee compared the original data files from W and E county and the first and second OVC. The reason for a discrepancy was determined individually for all non-concordant cases or breast cancer deaths. RESULTS: Of the 2615 cases included by the W-E Trial or the OVC, there were 478 (18%) disagreements. Of the disagreements 82% were due to inclusion/exclusion criteria, and 18% to disagreement with respect to cause of death or vital status at ascertainment. For E-County, the OVC inclusion rules and register based determination of cause of death (second OVC) rather than individual case review (W-E Trial and 1st OVC) resulted in a reduction of the estimate of the effect of screening, but for W-County the difference between the original trial and the OVC was modest. CONCLUSIONS: The conclusion that invitation to mammography screening reduces breast cancer mortality remains robust. Disagreements were mainly due to study design issues, while disagreements about cause of death were a minority. When secondary research does not adhere to the protocols of the primary research projects, the consequences of such design differences should be investigated and reported. Register linkage of trials can add follow-up information. The precision of trials with modest size is enhanced by individual monitoring of case status and outcome status such as determination of cause of death. PMID- 19564520 TI - Invitation management initiative to improve uptake of breast cancer screening in an urban UK Primary Care Trust. AB - OBJECTIVES: In an attempt to improve breast cancer screening uptake and coverage, persistent non-attenders in the Heart of Birmingham Teaching Primary Care Trust were included in an invitation management initiative. METHODS: Persistent non attenders were identified in routine screening lists. Phone contact was attempted or a home visit was made. If the case was not resolved, a second appointment was made and further phone calls and home visits were attempted. RESULTS: Of 548 persistent non-attenders identified, 228 (42%) declined screening, 171 (31%) attended, 72 (13%) had moved away or died, 11 (2%) were recently screened or under care for other conditions. Sixty-six cases (12%) remained unresolved. Fourteen women opted to be permanently withdrawn from the National Health Service Breast Screening Programme (NHSBSP). Twenty-four women had a negative experience of breast cancer screening (defaulted, recalled for assessment, recalled for technical reasons). No malignancies were found. A total of 1375 phone calls and 230 home visits were attempted. Uptake would have been 62.2% if none of the persistent non-attenders included in the initiative had attended for screening. With the initiative, uptake of breast cancer screening was increased to 65.3%. CONCLUSIONS: Phone calls and home visits resulted in only a moderate increase in breast cancer screening uptake. The initiative encouraged nervous attenders who were reassured about the screening process. However, more women declined screening than were screened and the initiative made it easier for women to request to be permanently withdrawn from the NHSBSP. PMID- 19564521 TI - Variation in the cervical cancer screening compliance among women with disability. AB - OBJECTIVE: To investigate the relationship between the level of disability and regular Pap smear testing among women in Taiwan and explore how this relationship may vary with the various levels of physician availability. METHODS: This population-based cohort study followed a total of 5,469,581 women from Taiwan, who were 30 years old or older in 2001 and covered the period January 2001 to December 2003. Of the total study population, 184,701 individuals were women with disability. Gynecologist-obstetrician/general practitioner to female population ratio was used as an indicator of physician availability. Multiple logistical regression models were used. RESULTS: After adjusting for age, socioeconomic status, racial group, residence area and physician availability, women with severe disability (OR = 0.38; 95% CI: 0.38, 0.39) were the least likely to undergo Pap smear testing. Women with moderate disability (OR = 0.59; 95% CI: 0.58, 0.60) and mild disability (OR = 0.88; 95% CI: 0.86, 0.89) were also significantly less likely to undergo a routine test than women without disability. Women residing in the areas with the greatest physician availability (OR = 0.93; 95% CI: 0.93, 0.94) were significantly less likely to undergo a Pap test than those in the areas with the lowest level of resource availability. The disparity in routine screening between women with and without disability remained across the different levels of physician availability. CONCLUSIONS: In Taiwan, women with disability were found to be at higher risk of lower compliance than women without disability. The gap between women with and without disability persisted across different levels of physician availability. PMID- 19564522 TI - Validity of self-reported Pap smear history in Norwegian women. AB - OBJECTIVES: To assess the validity of self-reported Papanicolau (Pap) smear history in Norwegian women and to identify characteristics that influence the validity. METHODS: Interview data from a sample of 16,574 Norwegian women, aged 18-45, in 2004-2005, was compared with information from the population-based cytology register. Crude validity in the self-reports with respect to ever/never having taken a Pap smear was summarized. The validity of the reported interval since last Pap smear was assessed by a smoothed distribution of the reported interval, stratified by the registered interval. Characteristics of influence on validity were identified by logistic regression for true positives (sensitivity and positive predictive value), true negatives (specificity and negative predictive value) and for more than one year discrepancy in time since last Pap smear, between reported and registered interval. RESULTS: Overall validity was summarized by: concordance = 0.9, sensitivity = 0.97, positive predictive value = 0.92, specificity = 0.55, negative predictive value = 0.78 and report-to-records ratio = 1.51. The variance in the reported interval increased proportionally with the registered interval, and women tended to underestimate the interval (telescoping). Age and registered number of years since last Pap smear had the strongest influence on ever/never and time interval validity, respectively. CONCLUSIONS: Estimated screening rates, based on self-reporting without organized screening, are biased. Telescoping leads to increased risk for developing invasive disease, because women will postpone their next Pap smear. PMID- 19564523 TI - Stage shift in PSA-detected prostate cancers - effect modification by Gleason score. AB - OBJECTIVE: This paper aims to investigate whether the stage shift (where more cancers are detected at an earlier stage) in prostate-specific antigen (PSA) detected cancers differs by Gleason score. METHODS: Between 2002 and 2005, 1514 men aged 50-69 years were identified with prostate cancer following community based PSA testing as part of the ProtecT study. In the same period, 2021 men aged 50-69 years with clinically diagnosed prostate cancer were registered at a population-based cancer registry in the East of England. Using logistic regression analysis and controlling for age, the odds ratio (OR) for advanced stage (TNM stage T3 and above) prostate cancer among the PSA-detected group was compared with the clinically diagnosed tumours. The evidence that stage shift differs by Gleason score was assessed using the likelihood ratio test for interaction. RESULTS: Advanced stage disease among the PSA-detected cancers was less common than among the clinically detected cancers (OR = 0.47, 95% CI 0.39 0.56). PSA-detected tumours had a substantial shift to earlier-stage disease where the Gleason score was <7 (OR = 0.52; 95% CI 0.36-0.77, P < 0.001) but showed no such shift where the Gleason score was 7 or more (OR = 0.84; 95% CI 0.66-1.07, P = 0.1). There was evidence of interaction between detection mode and Gleason score (P = 0.03). CONCLUSION: The observed stage shift could be partially explained by length bias or overdiagnosis. These findings may have implications on understanding pathways of prostate cancer progression and on identifying potential targets for screening, pending further investigation of complexities of associations between PSA testing, Gleason score, and stage. PMID- 19564524 TI - Adjustment of serum markers in first trimester screening. AB - First trimester combined screening can be performed using maternal serum pregnancy-associated plasma protein A, total human chorionic gonadotropin (hCG) and ultrasound measurement of nuchal translucency at 11-13 weeks of pregnancy. Our objective was to explore the effects of covariates on total hCG in the first trimester. First trimester total hCG levels were significantly increased in twins (median = 1.87 MoM), mildly increased in pregnancies achieved by in vitro fertilization (1.04 MoM) and decreased in smokers (0.80 MoM). PMID- 19564526 TI - Response to a novel multitargeted tyrosine kinase inhibitor pazopanib in metastatic Merkel cell carcinoma. PMID- 19564527 TI - Hodgkin's lymphoma with digital clubbing. PMID- 19564529 TI - Building a protocol expressway: the case of Mayo Clinic Cancer Center. AB - PURPOSE: Inconsistencies and errors resulting from nonstandard processes, together with redundancies, rework, and excess workload, lead to extended time frames for clinical trial protocol development. This results in dissatisfaction among sponsors, investigators, and staff and restricts the availability of novel treatment options for patients. METHODS: A team of experts from Mayo Clinic formed, including Protocol Development Unit staff and management from the three Mayo Clinic campuses (Florida, Minnesota, and Arizona), a systems and procedures analyst, a quality office analyst, and two physician members to address the identified deficiencies. The current-state process was intensively reviewed, and improvement steps were taken to accelerate the development and approval of cancer related clinical trials. The primary goal was to decrease the time from receipt of a new protocol through submission to an approving authority, such as the National Cancer Institute or institutional review board. RESULTS: Using the Define, Measure, Analyze, Improve, Control (DMAIC) framework infused with Lean waste-reduction methodologies, areas were identified for improvement, including enhancing first-time quality and processing new studies on a first-in/first-out basis. The project was successful in improving the mean turnaround time for internally authored protocols (P < .001) from 25.00 weeks (n = 41; range, 3.43 to 94.14 weeks) to 10.15 weeks (n = 14; range, 4.00 to 22.14 weeks). The mean turnaround time for externally authored protocols was improved (P < .001) from 20.61 weeks (n = 85; range, 3.29 to 108.57 weeks) to 7.79 weeks (n = 50; range, 2.00 to 20.86 weeks). CONCLUSION: DMAIC framework combined with Lean methodologies is an effective tool to structure the definition, planning, analysis, and implementation of significant process changes. PMID- 19564528 TI - African American women who receive primary anthracycline- and taxane-based chemotherapy for triple-negative breast cancer suffer worse outcomes compared with white women. PMID- 19564530 TI - Internal mammary node radiation: a proposed technique to spare cardiac toxicity. PMID- 19564531 TI - Postoperative surveillance recommendations for early stage colon cancer based on results from the clinical outcomes of surgical therapy trial. AB - PURPOSE: Intensive postoperative surveillance is associated with improved survival and recommended for patients with late stage (stage IIB and III) colon cancer. We hypothesized that stage I and IIA colon cancer patients would experience similar benefits. PATIENTS AND METHODS: Secondary analysis of data from the Clinical Outcomes of Surgical Therapy trial was performed by analyzing results according to TNM stage; early (stage I and IIA, 537 patients) and late (stage IIB and III, 254 patients) stage disease. Five-year recurrence rates were higher in patients with late (35.7%) versus early stage disease (9.5%). Early and late stage salvage rates, recurrence patterns and methods of first detection were compared by chi(2) test. RESULTS: Salvage rates for early- and late-stage disease patients with recurrence were the same (35.9% v 37%; P = .9, respectively). Median survival after second surgery after recurrence was 51.2 and 35.8 months for early- and late-stage patients, respectively. Single sites of first recurrence did not significantly differ between early and late stage, but multiple sites of recurrence occurred less often in early-stage patients (3.6% v 28.6%, for early v late, respectively; P < .001). METHODS of first detection of recurrence were not significantly different: carcinoembryonic antigen (29.1% v 37.4%), computed tomography scan (23.6% v 26.4%), chest x-ray (7.3% v 12.1%), and colonoscopy (12.7% v 8.8%), for early versus late stage disease, respectively. CONCLUSION: Patients with early-stage colon cancer have similar sites of recurrence, and receive similar benefit from postrecurrence therapy as late-stage patients; implementation of surveillance guidelines for early-stage patients is appropriate. PMID- 19564533 TI - Sensitivity to first-line chemotherapy for metastatic breast cancer in BRCA1 and BRCA2 mutation carriers. AB - PURPOSE: Preclinical as well as a few small retrospective, neoadjuvant studies suggest that breast cancer (cells) without functional BRCA1 or BRCA2 protein have an increased sensitivity to some chemotherapeutic agents causing double-strand DNA breaks. In this study we assessed the sensitivity to standard first-line chemotherapy of metastatic BRCA1/2-associated breast cancer, compared with sporadic breast cancer patients. PATIENTS AND METHODS: From the Family Cancer Clinic database, we selected 93 BRCA1- and 28 BRCA2-associated breast cancer patients treated with chemotherapy for metastatic disease before January 1, 2007. Objective response (OR), progression-free survival (PFS), and overall survival (OS) after start of first-line chemotherapy were compared with those of sporadic patients, matched for year of birth, age at diagnosis of primary breast cancer, and year of detection of metastatic disease. RESULTS: The chemotherapy regimens most frequently used were anthracycline-based (n = 147) and cyclophosphamide, methotrexate, and fluorouracil (CMF)/CMF like (n = 68). As compared to sporadic patients, BRCA2-associated patients had a significantly higher OR (89% v 50%; P = .001), a longer PFS (hazard ratio multivariate [HR(mult)] 0.64; P = .04) and a prolonged OS (HR(mult), 0.53; P = .005) after start of first-line chemotherapy for metastatic breast cancer. For BRCA1-associated patients, a nonsignificant trend for an increased OR (66% v 50%; P = .07), and a longer PFS (HR(mult), 0.79; P = .14) after first-line chemotherapy for metastatic breast cancer was observed, but not for OS. CONCLUSION: BRCA2-associated breast cancer is more sensitive to standard first-line chemotherapy for metastatic breast cancer in comparison with sporadic breast cancer, especially to anthracyclines. For BRCA1-associated breast cancer no statistically significant higher sensitivity was observed. PMID- 19564532 TI - Intensity-modulated radiation therapy with or without chemotherapy for nasopharyngeal carcinoma: radiation therapy oncology group phase II trial 0225. AB - PURPOSE: To investigate the feasibility of intensity-modulated radiation therapy (IMRT) with or without chemotherapy, and to assess toxicities, failure patterns, and survivals in patients with nasopharyngeal carcinoma (NPC). PATIENTS AND METHODS: Radiation consisted of 70 Gy given to the planning target volumes of primary tumor plus any N+ disease and 59.4 Gy given to subclinical disease, delivered over 33 treatment days. Patients with stage T2b or greater or with N+ disease also received concurrent cisplatin (100 mg/m(2)) on days 1, 22, and 43 followed by adjuvant cisplatin (80 mg/m(2)) on day 1; fluorouracil (1,000 mg/m(2)/d) on days 1 through 4 administered every 4 weeks for three cycles. Tumor, clinical status, and acute/late toxicities were assessed. The primary objective was to test the transportability of IMRT to a multi-institutional setting. RESULTS: Between February 2003 and November 2005, 68 patients with stages I through IVB NPC (of which 93.8% were WHO types 2 and 3) were enrolled. Prescribed IMRT (target delineation) was given to 83.8%, whereas 64.9% received chemotherapy per protocol. The estimated 2-year local progression-free (PF), regional PF, locoregional PF, and distant metastasis-free rates were 92.6%, 90.8%, 89.3%, and 84.7%, respectively. The estimated 2-year PF and overall survivals were 72.7% and 80.2%, respectively. Acute grade 4 mucositis occurred in 4.4%, and the worst late grade 3 toxicities were as follows: esophagus, 4.7%; mucous membranes, 3.1%; and xerostomia, 3.1%. The rate of grade 2 xerostomia at 1 year from start of IMRT was 13.5%. Only two patients complained of grade 3 xerostomia, and none had grade 4 xerostomia. CONCLUSION: It was feasible to transport IMRT with or without chemotherapy in the treatment of NPC to a multi institutional setting with 90% LRPF rate reproducing excellent reports from single institutions. Minimal grade 3 and lack of grade 4 xerostomia were encouraging. PMID- 19564535 TI - Phase II study of safety and efficacy of motesanib in patients with progressive or symptomatic, advanced or metastatic medullary thyroid cancer. AB - PURPOSE: This phase II study investigated the efficacy and tolerability of motesanib, an investigational, highly selective inhibitor of vascular endothelial growth factor receptors 1, 2, and 3; platelet-derived growth factor receptor; and Kit in advanced medullary thyroid cancer (MTC). PATIENTS AND METHODS: Patients with locally advanced or metastatic, progressive or symptomatic MTC received motesanib 125 mg/d orally for up to 48 weeks or until unacceptable toxicity or disease progression. The primary end point was objective response by independent review. Other end points included duration of response, progression-free survival, safety, pharmacokinetics, and changes in tumor markers. RESULTS: Of 91 enrolled patients who received motesanib, two (2%) achieved objective response (95% CI, 0.3% to 7.7%); their duration of response was 32 weeks (censored) and 21 weeks (disease progressed). Eighty-one percent of patients had stable disease (48% had durable stable disease > or = 24 weeks), 8% had disease progression as best response, and 9% were not evaluated; 76% experienced a decrease from baseline in target lesion measurement. Median progression-free survival was 48 weeks (95% CI, 43 to 56 weeks). Among patients with tumor marker analysis, 69 (83%) of 83 and 63 (75%) of 84 had decreased serum calcitonin and carcinoembryonic antigen during treatment, respectively, compared with baseline. The most common treatment-related adverse events were diarrhea (41%), fatigue (41%), hypothyroidism (29%), hypertension (27%), and anorexia (27%). In pharmacokinetic analyses, motesanib trough concentrations were lower compared with differentiated thyroid cancer patients from the same study. CONCLUSION: Although the objective response rate was low, a significant proportion of MTC patients (81%) achieved stable disease while receiving motesanib. PMID- 19564536 TI - A good life. PMID- 19564534 TI - Insulin resistance and risk factors for cardiovascular disease in young adult survivors of childhood acute lymphoblastic leukemia. AB - PURPOSE: To determine the prevalence of insulin resistance and other risk factors for cardiovascular disease (CVD) in young adult survivors of childhood acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: In this cross-sectional evaluation of 118 survivors of childhood ALL (median age, 23.0 years; range, 18 to 37 years), insulin resistance was estimated using the homeostasis model for assessment of insulin resistance (HOMA-IR). Sex-specific comparisons were made with a cohort of 30- to 37-year-old individuals from the same region participating in the Dallas Heart Study (DHS, N = 782). ALL survivors were stratified by treatment with and without cranial radiotherapy (CRT). RESULTS: Female ALL survivors had a significantly higher HOMA-IR (CRT, mean 4.6, 95% CI, 3.6 to 5.7; no CRT, mean 3.3, 95% CI, 2.8 to 3.8) in comparison with DHS women (mean 2.4, 95% CI, 2.2 to 2.7). Eighty percent of women treated with CRT had at least three of six CVD risk factors, and they were significantly more likely to have three or more risk factors compared with DHS women (odds ratio [OR], 5.96; 95% CI, 2.15 to 16.47). Male ALL survivors had a significantly higher HOMA-IR (CRT, mean 4.0, 95% CI, 2.8 to 5.6; no CRT, mean 3.4, 95% CI, 2.9 to 3.9) in comparison with DHS men (mean 2.3, 95% CI, 2.1 to 2.6), but were not more likely to have multiple CVD risk factors. CONCLUSION: ALL survivors had an increased prevalence of insulin resistance in comparison with a cohort of older individuals from the same community. Importantly, women treated with CRT seem to have an increased prevalence of multiple CVD risk factors, warranting close monitoring and risk-reducing strategies. PMID- 19564538 TI - Improving the methodologic and ethical validity of best supportive care studies in oncology: lessons from a systematic review. AB - PURPOSE: To systematically review the best supportive care (BSC) literature and to evaluate the ethical and methodologic validity issues by using widely acknowledged criteria. METHODS: Two search strings that included both cancer and supportive as terms (with random article type, or review or meta-analysis) explored databases from 1966 to 2008. Citations, abstracts, and papers were reviewed for inclusion criteria, and relevant data were extracted by two independent researchers. Data were validated for accuracy. Ethical and methodologic validity were evaluated by using the criteria derived from the Helsinki Requirements of the WMA; CONSORT statements for the evaluation of reports of randomized, controlled trials; and the universal requirements for ethical clinical research. RESULTS: Forty-three published papers were identified that described 32 studies, 20 of which incorporated the design of treatment plus supportive care (SC) versus SC alone, and 12 of which incorporated the design of treatment versus SC. Most of the studies had poor compliance to critical Helsinki requirements, to methodologic precautions derived from the CONSORT statement for studies involving a nonpharmacologic arm, and to four of seven universal requirements for ethical clinical research. CONCLUSION: Lack of rigor in BSC studies has contributed to a generation of research with widespread ethical and methodologic shortcomings. Ad hoc SC and lack of standardization of SC delivery may be sources of systematic bias or error in BSC trials. Rectifying these shortcomings in future studies demands greater vigilance toward these issues by researchers, institutional review boards, editors, and peer reviewers. Given the prevalence of overlooked problems that are later identified, currently open BSC studies should be reevaluated by institutional review boards and researchers to check for ethical and methodologic validity, and identified shortcomings should be addressed. PMID- 19564539 TI - Severe photosensitivity reaction to vandetanib. PMID- 19564540 TI - Exercise generates lactate and fluid intake: effects on mitochondrial function in heart and vascular smooth muscle. PMID- 19564541 TI - A novel amiloride-sensitive h+ transport pathway mediates enhanced superoxide production in thick ascending limb of salt-sensitive rats, not na+/h+ exchange. AB - It has been reported previously that H(+) efflux via the Na(+)/H(+) exchange stimulates NAD(P)H oxidase-dependent superoxide (O(2)(.-)) production in medullary thick ascending limb. We have demonstrated recently that N-methyl amiloride-sensitive O(2)(.-) production is enhanced in the thick ascending limb of Dahl salt-sensitive (SS) rats, suggesting that H(+) efflux through Na(+)/H(+) exchangers may promote renal oxidative stress and the development of hypertension in these animals. In the current study we demonstrate, using selective and potent inhibitors, that inhibition of Na(+)/H(+) exchange does not mediate the ability of N-methyl-amiloride to inhibit thick ascending limb O(2)(.-) production. To determine the mechanism of action of N-methyl-amiloride, we examined H(+) efflux and O(2)(.-) production in SS and SS.13(BN) thick ascending limbs of prehypertensive, 0.4% NaCl-fed rats. Tissue strips containing the medullary thick ascending limb were isolated from male SS and salt-resistant consomic SS.13(BN) rats, loaded with either dihydroethedium or 2',7'-bis-(2-carboxyethyl)-5-(and-6) carboxyfluorescein, acetoxymethyl ester, and imaged in a heated tissue bath. In Na(+)-replete media, activation of Na(+)/H(+) exchange using an NH(4)Cl prepulse did not stimulate thick ascending limb O(2)(.-) production. In Na(+)-free media containing BaCl(2) in which Na(+)/H(+) activity was inhibited, an NH(4)Cl prepulse stimulated O(2)(.-) production in medullary thick ascending limb renal tubular segments. This response was enhanced in medullary thick ascending limb of SS rats (slope Deltaethidium/Deltadihydroethedium=0.029+/-0.004) compared with SS.13(BN) rats (slope=0.010+/-0.004; P<0.04) and could be inhibited by N-methyl amiloride (slope=0.005+/-0.002 and 0.006+/-0.002 for SS and SS.13(BN), respectively). We concluded that only H(+) efflux through a specific, as-yet unidentified, amiloride-sensitive H(+) channel promotes O(2)(.-) production in the medullary thick ascending limb and that this channel is upregulated in SS rats. PMID- 19564542 TI - Role of pulse pressure amplification in arterial hypertension: experts' opinion and review of the data. PMID- 19564543 TI - Endothelial cells negatively modulate reactive oxygen species generation in vascular smooth muscle cells: role of thioredoxin. AB - In intact vessels, endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) act as an integrated system, possibly through reactive oxygen species (ROS). Using a coculture system we tested whether ECs modulate VSMC redox status by regulating activity of NAD(P)H oxidase and antioxidants. VSMC production of O(2)(*-), H(2)O(2), and NO was assessed using fluoroprobes and amplex-red. NAD(P)H oxidase subunit expression and oxidase activity were determined by Western blotting and chemiluminescence, respectively. Expression of thioredoxin, SOD, growth signaling pathways (PCNA, p21cip1, CDK4, ERK1/2, p38MAPK) was evaluated by immunoblotting. Thioredoxin activity was assessed by the insulin disulfide reduction assay. In cocultured conditions, VSMC ROS production was reduced by approximately 50% without changes in NAD(P)H oxidase expression/activity versus monoculture (P<0.05). This was associated with decreased cell growth (P<0.05). Expression of Cu/Zn SOD and thioredoxin was increased in coculture versus monoculture VSMCs (P<0.01). Pretreatment of ECs with L-NAME (NOS inhibitor), NS-398 (Cox2 inhibitor), and HET0016 (20-HETE inhibitor) did not influence VSMC ROS formation, whereas CDNB, thioredoxin reductase inhibitor, abolished ROS modulating effects of ECs. These findings indicate that in a coculture system recapitulating intact vessels, ECs negatively regulate ROS production in VSMCs through thioredoxin upregulation. Functionally this is associated with growth inhibition. The modulatory actions of ECs are independent of NOS/NO, Cox2, and HETE and do not involve NAD(P)H oxidase. Our data identify novel mechanisms whereby ECs protect against VSMC oxidative stress, a process that may be important in maintaining vascular integrity. PMID- 19564544 TI - Cardiac Renin levels are not influenced by the amount of resident mast cells. AB - To investigate whether mast cells release renin in the heart, we studied renin and prorenin synthesis by such cells, using the human mast cell lines human mastocytoma 1 and LAD2, as well as fresh mast cells from mastocytosis patients. We also quantified the contribution of mast cells to cardiac renin levels in control and infarcted rat hearts. Human mastocytoma 1 cells contained and released angiotensin I-generating activity, and the inhibition of this activity by the renin inhibitor aliskiren was comparable to that of recombinant human renin. Prorenin activation with trypsin increased angiotensin I-generating activity in the medium only, suggesting release but not storage of prorenin. The adenylyl cyclase activator forskolin, the cAMP analogue 8-db-cAMP, and the degranulator compound 48/80 increased renin release without affecting prorenin. Angiotensin II blocked the forskolin-induced renin release. Angiotensin I generating activity was undetectable in LAD2 cells and fresh mast cells. Nonperfused rat hearts contained angiotensin I-generating activity, and aliskiren blocked approximately 70% of this activity. A 30-minute buffer perfusion washed away >70% of the aliskiren-inhibitable angiotensin I-generating activity. Prolonged buffer perfusion or compound 48/80 did not decrease cardiac angiotensin I-generating activity further or induce angiotensin I-generating activity release in the perfusion buffer. Results in infarcted hearts were identical, despite the increased mast cell number in such hearts. In conclusion, human mastocytoma 1 cells release renin and prorenin, and the regulation of this release resembles that of renal renin. However, this is not a uniform property of all mast cells. Mast cells appear an unlikely source of renin in the heart, both under normal and pathophysiological conditions. PMID- 19564545 TI - Relentless progression toward sustained hypertension. PMID- 19564546 TI - Plasma-mediated vascular dysfunction in the reduced uterine perfusion pressure model of preeclampsia: a microvascular characterization. AB - Preeclampsia is associated with widespread maternal vascular dysfunction, which is thought to be mediated by circulating factor(s). The aim of the study was to characterize vascular function in the reduced uterine perfusion pressure (RUPP) rat model of preeclampsia and to investigate the role of plasma factors in mediating any observed changes in vascular reactivity. Mean arterial blood pressure and vascular function were measured in RUPP and control rats. Mesenteric vessels from both virgin and pregnant rats were exposed for 1 hour or overnight to plasma from both RUPP and control rats and their vascular function assessed. RUPP rats were characterized by severe hypertension, restricted fetal growth, and reduced placental weight (P<0.001). Vasorelaxation was impaired in resistance vessels from RUPP compared with control rats (acetylcholine: R(max) 70+/-3 versus 92+/-1 [NP] and 93+/-3% [sham], P<0.01; bradykinin: 40+/-2 versus 62+/-2 [NP] and 59+/-4% [sham], P<0.001). Incubation of vessels from pregnant (but not virgin) animals with RUPP plasma overnight resulted in an attenuation of vasorelaxant responses (acetylcholine: 63+/-7 versus 86+/-2%, P<0.05; bradykinin: 35+/-5 versus 55+/-6%, P<0.001). The residual relaxant response in RUPP plasma-treated vessels was not further attenuated after treatment with N(omega)-nitro-l-arginine methyl ester (acetylcholine: 57+/-7 versus 63+/-7%, ns; bradykinin: 37+/-5 versus 35+/-5%, ns). The RUPP rat model is characterized by an impaired response to vasodilators which may be attributable to one or more circulating factors. This plasma-mediated endothelial dysfunction appears to be a pregnancy-dependent effect. Furthermore, nitric oxide-mediated vasorelaxation appears to be absent in RUPP plasma-treated vessels. PMID- 19564547 TI - Effect of rosuvastatin on cardiac remodeling, function, and progression to heart failure in hypertensive heart with established left ventricular hypertrophy. AB - Hypertensive patients with left ventricular hypertrophy (LVH) are the most common high-risk group to develop heart failure with preserved ejection fraction. Recent reports have noted the favorable effect of statins on LVH. We evaluated the effect of rosuvastatin on cardiac remodeling, function, and progression to heart failure in a hypertensive rat model with established LVH. Dahl salt-sensitive rats were fed a high-salt diet until 13 weeks of age. After LVH was confirmed by echocardiography, rats were randomly assigned to control and statin treatment (n=18 each group). The statin-treated group was treated with rosuvastatin until 21 weeks of ages. Serial echocardiography, blood pressure monitoring, and miniaturized conductance catheter hemodynamic monitoring were performed at 21 weeks. Echocardiographic parameters were not significantly different between the groups. On hemodynamic monitoring, systolic performance parameters were similar between the groups, whereas end diastolic pressure-volume relationships were lower in the statin-treated group (0.014+/-0.008 versus 0.008+/-0.004 mm Hg/muL, P<0.05), suggesting improvement in myocardial stiffness. Pathological analysis showed attenuation of perivascular and interstitial fibrosis in the statin treated group (P<0.02). Rosuvastatin therapy did not alleviate LVH in hypertensive rats with established LVH, but it attenuated myocardial fibrosis and LV stiffness. It seems that rosuvastatin has limited therapeutic value when used to prevent progression from LVH to heart failure in hypertensive hearts. PMID- 19564548 TI - Long-term risk of sustained hypertension in white-coat or masked hypertension. AB - It is debated whether white-coat (WCHT) and masked hypertension (MHT) are at greater risk of developing a sustained hypertensive state (SHT). In 1412 subjects of the Pressioni Arteriose Monitorate e Loro Associazioni Study, we measured office blood pressure (BP), 24-hour ambulatory BP, and home BP. The condition of WCHT was identified as office BP >140/90 mm Hg and 24-hour BP mean <125/79 mm Hg or home BP <132/82 mm Hg. Corresponding values for MHT diagnosis were office BP <140/90 mm Hg, 24-hour BP > or =125/79 mm Hg, and home BP >or =132/82 mm Hg. SHT was identified when both office and 24-hour BP means or home BP were over threshold values and normotension was under the threshold value. Subjects were reassessed 10 years later to evaluate the BP status of the various conditions defined previously. At the first examination, 758 (54.1%), 225 (16.1%), 124 (8.9%), and 293 (20.9%) subjects were normotensive, WCHT, MHT, and SHT subjects, respectively. At the second examination, 136 normotensives (18.2%), 95 WCHT (42.6%), and 56 MHT (47.1%) subjects became SHT. As compared with normotensives, adjusting for age and sex, the risk of becoming SHT was significantly higher for WCHT and MHT subjects (odds ratio: 2.51 and 1.78, respectively; P<0.0001). Similar results were obtained when the definition of the various conditions was based on home BP. Independent contributors of worsening of hypertension status were not only baseline BP, but also, although to a lesser extent, metabolic variables and age. Subjects with WCHT and MHT are at increased risk of developing SHT. This may contribute to their prognosis that appears to be worse as compared with that of normotensive subjects. PMID- 19564549 TI - Growth arrest specific protein 6 participates in DOCA-induced target-organ damage. AB - Growth arrest-specific protein 6 (Gas 6) is involved in inflammatory kidney diseases, vascular remodeling, cell adhesion, and thrombus formation. We explored a role for Gas 6 in aldosterone-induced target organ damage. We observed that Gas 6 was upregulated in rats with high aldosterone levels. Mineralocorticoid receptor blockade prevented target organ damage and decreased the elevated Gas 6 expression. Vascular smooth muscle cells given aldosterone increased their Gas 6 expression in vitro. To test the pathophysiological relevance, we investigated the effects of deoxycorticosterone acetate (DOCA) on Gas 6 gene-deleted ((-/-)) mice. After 6 weeks DOCA, Gas 6(-/-) mice developed similar telemetric blood pressure elevations compared to wild-type mice but were protected from cardiac hypertrophy. Cardiac expression of interleukin 6 and collagen IV was blunted in Gas 6(-/-) mice, indicating reduced inflammation and fibrosis. Gas 6(-/-) mice also had an improved renal function with reduced albuminuria, compared to wild type mice. Renal fibrosis and fibronectin deposition in the kidney were also reduced. Gas 6 deficiency reduces the detrimental effects of aldosterone on cardiac and renal remodeling independent of blood pressure reduction. Gas 6 appears to play a role in mineralocorticoid receptor-mediated target organ damage. Furthermore, because warfarin interferes with Gas 6 protein expression, the findings could be of clinical relevance for anticoagulant choices. PMID- 19564550 TI - Thiazide effects and adverse effects: insights from molecular genetics. PMID- 19564551 TI - Change in blood pressure and incident dementia: a 32-year prospective study. AB - Studies of the association of high blood pressure (BP) with dementia are not consistent. Understanding long-term trajectories in blood pressure of those who do and do not develop dementia can help clarify the issue. The Honolulu Heart Program/Honolulu-Asia Aging Study followed a cohort of Japanese American men for an average of 32 years, with systolic BP (SBP) and diastolic BP (DBP) measured at 6 examinations and dementia assessed at the final 3. In an analysis of 1890 men who completed all 6 of the exams, 112 diagnosed with incident dementia at examination 6 were compared with the 1778 survivors without dementia. Trajectories in SBP and DBP up to and including the sixth examination were estimated with a repeated-measures analysis using 3 splines. From midlife to late life, men who went on to develop dementia had an additional age-adjusted increase in SBP of 0.26 mm Hg (95% CI: 0.01 to 0.51 mm Hg) per year compared with survivors without dementia. Over the late-life examinations, this group had an additional age-adjusted decline in SBP of 1.36 mm Hg (95% CI: 0.64 to 2.07 mm Hg) per year. These associations were strongest for vascular dementia and were reduced substantially in men who were previously taking antihypertensive medication. Similar changes in diastolic BP were observed, but only for vascular dementia, and the findings were not modified by antihypertensive treatment. Over a 32-year period, compared with men who did not, those who did develop dementia had a greater increase, followed by a greater decrease, in SBP. Both of these trends are modified by antihypertensive therapy. PMID- 19564552 TI - Prevention of pulmonary hypertension by Angiotensin-converting enzyme 2 gene transfer. AB - In spite of recent advancements in the treatment of pulmonary hypertension, successful control has yet to be accomplished. The abundant presence of angiotensin-converting enzyme 2 (ACE2) in the lungs and its impressive effect in the prevention of acute lung injury led us to test the hypothesis that pulmonary overexpression of this enzyme could produce beneficial outcomes against pulmonary hypertension. Monocrotaline (MCT) treatment of mice for 8 weeks resulted in significant increases in right ventricular systolic pressure, right ventricle:left ventricle plus septal weight ratio, and muscularization of pulmonary vessels. Administration of a lentiviral vector containing ACE2, 7 days before MCT treatment prevented the increases in right ventricular systolic pressure (control: 25+/-1 mm Hg; MCT: 44+/-5 mm Hg; MCT+ACE2: 26+/-1 mm Hg; n=6; P<0.05) and right ventricle:left ventricle plus septal weight ratio (control: 0.25+/-0.01; MCT: 0.31+/-0.01; MCT+ACE2: 0.26+/-0.01; n=8; P<0.05). A significant attenuation in muscularization of pulmonary vessels induced by MCT was also observed in animals overexpressing ACE2. These beneficial effects were associated with an increase in the angiotensin II type 2 receptor:angiotensin II type 1 receptor mRNA ratio. Also, pulmonary hypertension-induced increases in proinflammatory cytokines were significantly attenuated by lentiviral vector containing ACE2 treatment. Furthermore, ACE2 gene transfer in mice after 6 weeks of MCT treatment resulted in a significant reversal of right ventricular systolic pressure. These observations demonstrate that ACE2 overexpression prevents and reverses right ventricular systolic pressure and associated pathophysiology in MCT-induced pulmonary hypertension by a mechanism involving a shift from the vasoconstrictive, proliferative, and fibrotic axes to the vasoprotective axis of the renin-angiotensin system and inhibition of proinflammatory cytokines. PMID- 19564553 TI - Is it time to treat depression in patients with cardiovascular disease? PMID- 19564554 TI - Diabetic monocyte and vascular endothelial growth factor signaling impairment. PMID- 19564555 TI - Drug-eluting stents in animals and patients: where do we stand today? PMID- 19564556 TI - Preoperative hemoglobin level as a predictor of survival after coronary artery bypass grafting: a comparison with the matched general population. AB - BACKGROUND: The predictive value of the preoperative hemoglobin value after coronary artery bypass grafting (CABG) has not been well established. We studied how the preoperative hemoglobin level affects the survival of patients after CABG. Late mortality was compared with that of a general population. METHODS AND RESULTS: Early and late mortality of all consecutive patients undergoing isolated CABG between January 1998 and December 2007 were determined. Patients were classified into 4 groups stratified by preoperative hemoglobin level. The cutoff point for anemia was 13 g/dL for men and 12 g/dL for women. Expected survival of a matched general Dutch population cohort was obtained from the database of the Dutch Central Bureau for Statistics. After the exclusion of 122 patients who were lost to follow-up and 481 patients with missing preoperative hemoglobin levels, complete data were obtained in 10,025 patients. Multivariate logistic regression analyses revealed anemia to be an independent risk factor for higher early mortality. Cox regression analyses revealed low hemoglobin level, both as a continuous variable and as a dichotomous variable (anemia), to be a predictor of higher late mortality. Compared with expected survival, patients with the lowest preoperative hemoglobin levels had a worse outcome, whereas patients with the highest hemoglobin levels had a better outcome. CONCLUSIONS: A lower preoperative hemoglobin level is an independent predictor of late mortality in patients undergoing CABG, whereas anemia is a risk factor for early and late mortality. Compared with the general population, anemic patients had worse survival than expected, whereas nonanemic patients had better survival than expected. PMID- 19564557 TI - Elevated depression symptoms predict long-term cardiovascular mortality in patients with atrial fibrillation and heart failure. AB - BACKGROUND: Depression predicts prognosis in many cardiac conditions, including congestive heart failure (CHF). Despite heightened cardiac risk in patients with comorbid atrial fibrillation (AF) and CHF, depression has not been studied in this group. This substudy, from the AF-CHF Trial of rate- versus rhythm-control strategies, investigated whether depression predicts long-term cardiovascular mortality in patients with left ventricular ejection fraction or=14). Over a mean follow-up of 39 months, there were 246 cardiovascular deaths (111 presumed arrhythmic; 302 all-cause deaths). Cox proportional hazards models adjusted for other prognostic factors (including age, marital status, cause of CHF, creatinine level, left ventricular ejection fraction, paroxysmal AF, previous AF hospitalization, previous electrical conversion, and baseline medications) showed that elevated depression scores significantly predicted cardiovascular mortality (primary outcome), arrhythmic death, and all-cause mortality. The adjusted hazard ratios were 1.57 (95% confidence interval 1.20 to 2.07, P<0.001), 1.69 (95% confidence interval 1.13 to 2.53, P=0.01), and 1.38 (95% confidence interval 1.07 to 1.77, P=0.01), respectively. The risks associated with depression and marital status were additive, with the highest risk in depressed patients who were unmarried. CONCLUSIONS: Elevated depression symptoms are related to cardiovascular mortality even after adjustment for other prognostic indicators in patients with comorbid AF and CHF who receive optimized treatment. Unmarried patients are also at increased risk. Mechanisms and treatment options deserve additional study. PMID- 19564558 TI - Lecithin: cholesterol acyltransferase expression has minimal effects on macrophage reverse cholesterol transport in vivo. AB - BACKGROUND: Lecithin:cholesterol acyltransferase (LCAT) catalyzes the formation of plasma cholesteryl ester, plays a key role in high-density lipoprotein metabolism, and has been believed to be critical in the process of reverse cholesterol transport (RCT). METHODS AND RESULTS: The role of LCAT in RCT from macrophages was quantified with a validated assay involving intraperitoneal injection in mice of (3)H-cholesterol-labeled J774 macrophages and monitoring the appearance of tracer in plasma, liver, bile, and feces. Human LCAT overexpression in human apolipoprotein A-I transgenic mice substantially increased plasma high density lipoprotein cholesterol levels but surprisingly did not increase macrophage RCT. Even in the setting of coexpression of scavenger receptor BI or cholesteryl ester transfer protein, both of which promoted the transfer of LCAT derived high-density lipoprotein cholesterol ester to the liver, LCAT overexpression still had no effect on RCT. Serum from LCAT-overexpressing mice had reduced ability to promote cholesterol efflux from macrophages ex vivo via ABCA1. To determine the effect of LCAT deficiency on macrophage RCT, LCAT(-/-) and LCAT(+/-) mice were compared with wild-type mice. Despite extremely low plasma levels of high-density lipoprotein cholesterol, LCAT-deficient mice had only a 50% reduction in RCT. LCAT(+/-) mice had normal RCT despite a significant reduction in high-density lipoprotein cholesterol. Serum from LCAT-deficient mice had increased ability to promote ABCA1-mediated cholesterol efflux from macrophages ex vivo. CONCLUSIONS: These results demonstrate that LCAT overexpression does not promote an increased rate of macrophage RCT. Although LCAT activity does become rate limiting in the context of complete LCAT deficiency, RCT is reduced by only 50% even in the absence of LCAT. These data suggest that macrophage RCT may not be as dependent on LCAT activity as has previously been believed. PMID- 19564559 TI - Diabetes mellitus activates signal transduction pathways resulting in vascular endothelial growth factor resistance of human monocytes. AB - BACKGROUND: Monocytes are cellular components of wound repair, arteriogenesis, and atherogenesis. Vascular endothelial growth factor (VEGF)-A and placental growth factor recruit monocytes to sites of arteriogenesis via stimulation of VEGF receptor-1 (VEGFR-1). The chemotactic response of monocytes to VEGF-A is attenuated in individuals with diabetes mellitus (DM). This VEGF resistance correlates with impaired collateral growth. The aim of this study is to elucidate the molecular basis of VEGF resistance and impaired monocyte response in DM. METHODS AND RESULTS: Phosphorylation of Akt, p38, and extracellular signal regulated kinase 1/2 (ERK1/2) could be stimulated with either placental growth factor-1 or VEGF-A in monocytes from non-DM but not DM individuals. In contrast, formyl-methionyl-leucyl-phenylalanine caused a comparable activation of these molecules in both DM and non-DM monocytes. Baseline phosphorylation of Akt, p38, and ERK1/2 was significantly elevated in monocytes from DM compared with non-DM subjects. Of note, H(2)O(2) activated Akt, p38, and ERK1/2 in non-DM monocytes ex vivo. Protein tyrosine phosphatases had stronger oxidative modifications in monocytes from DM than from non-DM individuals, which reflects functional protein tyrosine phosphatase inhibition, similar to that seen after H(2)O(2) challenge. Overall, protein tyrosine phosphatase and protein tyrosine phosphatase-1B activity were reduced in DM monocytes. DM monocytes revealed higher expression of the receptor for advanced glycation end products. Stimulation with advanced glycation end products ligands resulted in activation of non-DM monocytes and inhibition of VEGFR-1-mediated chemotaxis. The elevated baseline phosphorylation/activation of Akt, p38, and ERK1/2 in DM monocytes likely causes the resistance to further stimulation with specific stimuli such as VEGF-A, revealing a molecular explanation of the DM-related signal transduction defect. CONCLUSIONS: We propose that elevated advanced glycation end products expression and increased oxidative stress in diabetic monocytes lead to activation of VEGFR 1-related signaling pathways and to desensitization of VEGFR-1 responses. These data establish VEGF resistance as a novel molecular concept for DM-related cellular dysfunction. PMID- 19564560 TI - Psychosocial modulators of angina response to myocardial ischemia. AB - BACKGROUND: Although angina is often caused by atherosclerotic obstruction of the coronary arteries, patients with similar amounts of myocardial ischemia may vary widely in their symptoms. We sought to compare clinical and psychosocial characteristics associated with more frequent angina after adjusting for the amount of inducible ischemia. METHODS AND RESULTS: From 2004 to 2006, 788 consecutive patients undergoing single-photon emission computed tomography stress perfusion imaging at 2 Seattle hospitals were assessed for their frequency of angina over the previous 4 weeks with the Seattle Angina Questionnaire and for a broad range of psychosocial characteristics. Among patients with demonstrable ischemia on single-photon emission computed tomography (summed difference score >or=2; n=191), angina frequency was categorized as none (Seattle Angina Questionnaire score=100; n=68), monthly (score=61 to 99; n=66), and weekly or daily (score=0 to 60; n=57). Using multivariable ordinal logistic regression, increasing angina was significantly associated with a history of coronary revascularization (odds ratio 2.24, 95% confidence interval 1.19 to 4.19), anxiety (odds ratio 4.72, 95% confidence interval 1.91 to 11.66), and depression (odds ratio 3.12, 95% confidence interval 1.45 to 6.69) after adjustment for the amount of inducible ischemia. CONCLUSIONS: Among patients with a similar burden of inducible ischemia, a history of coronary revascularization and current anxiety and depressive symptoms were associated with more frequent angina. These results support the study of angina treatment strategies that aim to reduce psychosocial distress in conjunction with efforts to lessen myocardial ischemia. PMID- 19564561 TI - High-density substrate mapping in Brugada syndrome: combined role of conduction and repolarization heterogeneities in arrhythmogenesis. AB - BACKGROUND: Two principal mechanisms are thought to be responsible for Brugada syndrome (BS): (1) right ventricular (RV) conduction delay and (2) RV subepicardial action potential shortening. This in vivo high-density mapping study evaluated the conduction and repolarization properties of the RV in BS subjects. METHODS AND RESULTS: A noncontact mapping array was positioned in the RV of 18 BS patients and 20 controls. Using a standard S(1)-S(2) protocol, restitution curves of local activation time and activation recovery interval were constructed to determine local maximal restitution slopes. Significant regional conduction delays in the anterolateral free wall of the RV outflow tract of BS patients were identified. The mean increase in delay was 3-fold greater in this region than in control (P=0<0.001). Local activation gradient was also maximally reduced in this area: 0.33+/-0.1 (mean+/-SD) mm/ms in BS patients versus 0.51+/ 0.15 mm/ms in controls (P<0.0005). The uniformity of wavefront propagation as measured by the square of the correlation coefficient, r(2), was greater in BS patients versus controls (0.94+/-0.04 versus 0.89+/-0.09 [mean+/-SD]; P<0.05). The odds ratio of BS hearts having any RV segment with maximal restitution slope >1 was 3.86 versus controls. Five episodes of provoked ventricular tachycardia arose from wave breaks originating from RV outflow tract slow-conduction zones in 5 BS patients. CONCLUSIONS: Marked regional endocardial conduction delay and heterogeneities in repolarization exist in BS. Wave break in areas of maximal conduction delay appears to be critical in the initiation and maintenance of ventricular tachycardia. These data indicate that further studies of mapping BS to identify slow-conduction zones should be considered to determine their role in spontaneous ventricular arrhythmias. PMID- 19564562 TI - Comparison of inflammatory response after implantation of sirolimus- and paclitaxel-eluting stents in porcine coronary arteries. AB - BACKGROUND: Although both sirolimus (CYPHER) and paclitaxel (TAXUS) drug-eluting stents have demonstrated efficacy and safety in clinical trials, human autopsy data have raised concerns about long-term healing and the potential for local inflammatory reactions. METHODS AND RESULTS: Overlapping stents (CYPHER drug eluting stents, Bx SONIC bare metal stents, TAXUS drug-eluting stents, and Liberte bare metal stents) were implanted in noninjured coronary arteries of 58 domestic swine. Histopathological evaluation of proximal, overlapped, and distal stented segments was determined with emphasis on inflammation at 30, 90, and 180 days. Circumferential granulomatous inflammation in all stented segments was defined as inflammation consisting of macrophages, multinucleated giant cells, lymphocytes, and granulocytes, including many eosinophils, adjacent to almost all struts. Circumferential granulomatous inflammation was more prevalent in CYPHER (9 of 23, 39%) compared with TAXUS (1 of 21, 5%; P=0.01) and control bare metal stents (0 of 44) in the combined 90- and 180-day cohorts. Only CYPHER specimens showed marked adventitial inflammation (P=0.0025) and fibrosis (P=0.0055) accompanied by extensive remodeling. Fibrin deposition within neointima and medial smooth muscle cell death were greater (both P<0.001) in TAXUS than CYPHER at 30 days, with more fibrin in TAXUS than CYPHER through 90 days (P<0.05). CONCLUSIONS: Although these data cannot be directly extrapolated to humans, the high prevalence in this porcine model of diffuse granulomatous inflammation seen with CYPHER stents, persisting at 180 days and associated with extensive remodeling of the artery, and persistent para-strut fibrin deposition with TAXUS stents emphasize the need for further investigation of biocompatibility with these and other novel combination drug/polymer drug-eluting stents. PMID- 19564563 TI - How much is life worth: cetuximab, non-small cell lung cancer, and the $440 billion question. AB - The spiraling cost of cancer care, in particular the cost of cancer therapeutics that achieve only marginal benefits, is under increasing scrutiny. Although health-care professionals avoid putting a value on a life, our limited resources require that society address what counts as a benefit, the extent to which cost should factor in deliberations, and who should be involved in these decisions. Professional societies, such as the American Society of Clinical Oncology, government agencies, including the Food and Drug Administration, and insurance companies should be involved. However, no segment of society is better qualified to address these issues than the oncology community. Oncologists must offer clear guidance for the conduct of research, interpretation of results, and prescription of chemotherapies. We review recent drug approvals and clinical trials and comment on their relevance to the issue of the spiraling cost of oncology therapeutics. We suggest some standards that would serve as a starting point for addressing these issues. PMID- 19564564 TI - Images in cardiovascular medicine. Cardiac involvement in Erdheim-Chester disease: magnetic resonance and computed tomographic scan imaging in a monocentric series of 37 patients. PMID- 19564565 TI - Letter by Jeilan et al regarding article, "Longitudinal strain delay index by speckle tracking imaging: a new marker of response to cardiac resynchronization therapy". PMID- 19564566 TI - Letter by Rosenstein and Parra regarding article, "Greater clinical benefit of more intensive oral antiplatelet therapy with prasugrel in patients with diabetes mellitus in the Trial to Assess Improvement in Therapeutic Outcomes by Optimizing Platelet Inhibition with Prasugrel: Thrombolysis in Myocardial Infarction 38". PMID- 19564568 TI - Acute valvular regurgitation. PMID- 19564569 TI - Images in cardiovascular medicine. Lung perfusion blood volume computed tomographic images of pulmonary embolism: before and after thrombolysis. PMID- 19564570 TI - DAMPs and inflammatory processes: the role of redox in the different outcomes. AB - Inflammation is deeply entangled with redox modulation. Triggering of PRRs on inflammatory cells induces ROS generation. As a consequence, activated cells mount antioxidant responses to counteract the possible harmful effects of oxidation. Therefore, when repair is completed, homeostasis is restored. Here, we describe some recent results showing that an exuberant antioxidant response to pro-oxidant inflammatory stimuli modifies not only the intra- but also the extracellular redox and contributes to the outcome of the inflammatory process. In particular, the role of redox modulation in IL-1beta secretion, in B lymphocyte differentiation to plasma cells, and in tumor progression will be discussed, and the potential consequences of extracellular redox alterations on DAMP activity will be considered. PMID- 19564571 TI - The antihypoxia-adenosinergic pathogenesis as a result of collateral damage by overactive immune cells. AB - Here, we attract attention to the possibility of iatrogenic exacerbation of immune-mediated tissue damage as a result of the unintended weakening of the tissue-protecting, hypoxia-adenosinergic pathway. These immunosuppressive, anti inflammatory pathways play a critical and nonredundant role in the protection of normal tissues from collateral damage during an inflammatory response. We believe that it is the tissue hypoxia associated with inflammatory damage that leads to local inhibition of overactive immune cells by activating A2AR and A2BR and stabilizing HIF-1alpha. We show in an animal model of acute lung injury that oxygenation (i.e., inspiring supplemental oxygen) reverses tissue hypoxia and exacerbates ongoing inflammatory lung tissue damage. However, little has been done to carefully investigate and prevent this in a clinical setting. Similarly, the consumption of caffeine antagonizes A2ARs, resulting in exacerbation of ongoing acute inflammation. It is suggested that although the elimination of hypoxia-adenosinergic immunosuppression is desirable to improve vaccines, it is important to take into account the unintentional effects of supplemental oxygen and caffeine, which may increase collateral, inflammatory tissue damage. PMID- 19564572 TI - The alarmin HMGB1 acts in synergy with endogenous and exogenous danger signals to promote inflammation. AB - The nuclear protein HMGB1 has previously been demonstrated to act as an alarmin and to promote inflammation upon extracellular release, yet its mode of action is still not well defined. Access to highly purified HMGB1 preparations from prokaryotic and eukaryotic sources enabled studies of activation of human PBMC or synovial fibroblast cultures in response to HMGB1 alone or after binding to cofactors. HMGB1 on its own could not induce detectable IL-6 production. However, strong enhancing effects on induction of proinflammatory cytokine production occurred when the protein associated with each of the separate proinflammatory molecules, rhIL-1beta, the TLR4 ligand LPS, the TLR9 ligand CpG-ODN, or the TLR1 TLR2 ligand Pam3CSK4. The bioactivities were recorded in cocultures with preformed HMGB1 complexes but not after sequential or simultaneous addition of HMGB1 and the individual ligands. Individual A-box and B-box domains of HMGB1 had the ability to bind LPS and enhance IL-6 production. Heat denaturation of HMGB1 eliminated this enhancement. Cocultures with HMGB1 and other proinflammatory molecules such as TNF, RANKL, or IL-18 did not induce enhancement. HMGB1 thus acts broadly with many but not all immunostimulatory molecules to amplify their activity in a synergistic manner. PMID- 19564573 TI - TLR4-mediated Cox-2 expression increases intestinal ischemia/reperfusion-induced damage. AB - Mesenteric IR induces significant inflammation and immune-mediated mucosal damage. TLR4 is a critical receptor in the induction of the inflammatory response and plays a role in intestinal homeostasis. To determine the role of TLR4 in IR induced epithelial damage, we performed IR studies using TLR4(lps-def) and TLR4(lps-n) mice and analyzed mucosal damage and inflammation. We found that the absence of TLR4 or TLR4-induced signaling attenuated local mucosal damage with significantly decreased cytokine and eicosanoid secretion including PGE2 production. Similar results were seen in MyD88-/- mice. Wild-type mice treated with NS-398 (a Cox-2 inhibitor) not only decreased PGE2 production but also attenuated tissue damage. In contrast, PGE2 was not sufficient to induce damage in the TLR4(lps-def) mice. Together, these data indicate that TLR4 stimulation of Cox-2 activation of PGE2 production is necessary but not sufficient for intestinal IR-induced damage and inflammation. PMID- 19564574 TI - Protein disulfide isomerase (PDI) associates with NADPH oxidase and is required for phagocytosis of Leishmania chagasi promastigotes by macrophages. AB - PDI, a redox chaperone, is involved in host cell uptake of bacteria/viruses, phagosome formation, and vascular NADPH oxidase regulation. PDI involvement in phagocyte infection by parasites has been poorly explored. Here, we investigated the role of PDI in in vitro infection of J774 macrophages by amastigote and promastigote forms of the protozoan Leishmania chagasi and assessed whether PDI associates with the macrophage NADPH oxidase complex. Promastigote but not amastigote phagocytosis was inhibited significantly by macrophage incubation with thiol/PDI inhibitors DTNB, bacitracin, phenylarsine oxide, and neutralizing PDI antibody in a parasite redox-dependent way. Binding assays indicate that PDI preferentially mediates parasite internalization. Bref-A, an ER-Golgi-disrupting agent, prevented PDI concentration in an enriched macrophage membrane fraction and promoted a significant decrease in infection. Promastigote phagocytosis was increased further by macrophage overexpression of wild-type PDI and decreased upon transfection with an antisense PDI plasmid or PDI siRNA. At later stages of infection, PDI physically interacted with L. chagasi, as revealed by immunoprecipitation data. Promastigote uptake was inhibited consistently by macrophage preincubation with catalase. Additionally, loss- or gain-of-function experiments indicated that PMA-driven NADPH oxidase activation correlated directly with PDI expression levels. Close association between PDI and the p22phox NADPH oxidase subunit was shown by confocal colocalization and coimmunoprecipitation. These results provide evidence that PDI not only associates with phagocyte NADPH oxidase but also that PDI is crucial for efficient macrophage infection by L. chagasi. PMID- 19564575 TI - The functional significance behind expressing two IL-8 receptor types on PMN. AB - PMN are critical to innate immunity and are fundamental to antibacterial defense. To localize to sites of infection, PMN possess receptors that detect chemoattractant stimuli elicited at the site, such as chemokines, complement split products, or bioactive lipids. Signaling through these receptors stimulates chemotaxis toward the site of infection but also activates a number of biochemical processes, with the result that PMN kill invading bacteria. PMN possess two receptors, CXCR1 and CXCR2, for the N-terminal ELR motif-containing CXC chemokines, although only two chemokine members bind both receptors and the remainder binding only CXCR2. This peculiar pattern in receptor specificity has drawn considerable interest and investigation into whether signaling through each receptor might impart unique properties on the PMN. Indeed, at first glance, CXCR1 and CXCR2 appear to be functionally redundant; however, there are differences. Considering these proinflammatory activities of activating PMN through chemokine receptors, there has been great interest in the possibility that blocking CXCR1 and CXCR2 on PMN will provide a therapeutic benefit. The literature examining CXCR1 and CXCR2 in PMN function during human and modeled diseases will be reviewed, asking whether the functional differences can be perceived based on alterations in the role PMN play in these processes. PMID- 19564576 TI - Epac inhibits apoptosis of human leukocytes. AB - cAMP is known to participate in the regulation of apoptosis in leukocytes. Depending on the cell type, pro- and antiapoptotic effects of cAMP have been described. Thus far, most of the cAMP-dependent effects have been attributed to the activation of PKA. However, Epac proteins (direct cAMP targets and guanine nucleotide exchange factors for Ras-like GTPases) have been shown recently to contribute to cAMP-dependent regulation of apoptosis. Therefore, we investigated the effects of the selective Epac activators 8-pCPT and Sp on apoptosis in human leukocytic cells (U937, HL-60, primary human mononuclear cells). We report here that Epac activation inhibits leukocyte apoptosis significantly. PMID- 19564577 TI - Clinical Reasoning: a 22-year-old woman with headache and diplopia. PMID- 19564578 TI - Teaching Video NeuroImages: positional exophthalmos in orbital varices. PMID- 19564580 TI - Progress report and a farewell. PMID- 19564581 TI - Natural history of Ullrich congenital muscular dystrophy. AB - OBJECTIVE: To describe the course, complications, and prognosis of Ullrich congenital muscular dystrophy (UCMD), with special reference to life-changing events, including loss of ambulation, respiratory insufficiency, and death. METHODS: Review of the case notes of 13 patients with UCMD, aged 15 years or older at last visit, followed up at a tertiary neuromuscular centre, London, UK, from 1977 to 2007. Data collected were age at onset of symptoms, presenting symptoms, mobility, contractures, scoliosis, skin abnormalities, respiratory function, and feeding difficulties. RESULTS: The mean age at onset of symptoms was 12 months (SD 14 months). Eight patients (61.5%) acquired independent ambulation at a mean age of 1.7 years (SD 0.8 years). Nine patients (69.2%) became constant wheelchair users at a mean age of 11.1 years (SD 4.8 years). Three patients continued to ambulate indoors with assistance. Forced vital capacity (FVC) values were abnormal in all patients from age 6 years. The mean FVC (% predicted) declined at a mean rate of 2.6% (SD 4.1%) yearly. Nine patients (69.2%) started noninvasive ventilation at a mean age of 14.3 years (SD 5.0 years). Two patients died of respiratory insufficiency. CONCLUSION: In Ullrich congenital muscular dystrophy (UCMD), the decline in motor and respiratory functions is more rapid in the first decade of life. The deterioration is invariable, but not always correlated with age or severity at presentation. This information should be of help to better anticipate the difficulties encountered by patients with UCMD and in planning future therapeutic trials in this condition. PMID- 19564582 TI - Incidence and prevalence of CIDP and the association of diabetes mellitus. AB - BACKGROUND: The reported prevalence of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) varies greatly, from 1.9 to 7.7 per 100,000. CIDP is reported to occur more commonly in patients with diabetes mellitus (DM) but has not been rigorously tested. OBJECTIVES: To determine the incidence (1982 2001) and prevalence (on January 1, 2000) of CIDP in Olmsted County, Minnesota, and whether DM is more frequent in CIDP. METHODS: CIDP was diagnosed by clinical criteria followed by review of electrophysiology. Cases were coded as definite, probable, or possible. DM was ascertained by clinical diagnosis or current American Diabetes Association glycemia criteria. RESULTS: One thousand five hundred eighty-one medical records were reviewed, and 23 patients (10 women and 13 men) were identified as having CIDP (19 definite and 4 probable). The median age was 58 years (range 4-83 years), with a median disease duration at diagnosis of 10 months (range 2-64 months). The incidence of CIDP was 1.6/100,000/year. The prevalence was 8.9/100,000 persons on January 1, 2000. Only 1 of the 23 CIDP patients (4%) also had DM, whereas 14 of 115 age- and sex-matched controls (12%) had DM. CONCLUSIONS: 1) The incidence (1.6/100,000/year) and prevalence (8.9/100,000) of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) are similar to or higher than previous estimates. 2) The incidence of CIDP is similar to that of acute inflammatory demyelinating polyradiculoneuropathy within the same population. 3) Diabetes mellitus (DM) is unlikely to be a major risk covariate for CIDP, but we cannot exclude a small effect. 4) The perceived association of DM with CIDP may be due to misclassification of other forms of diabetic neuropathies and excessive emphasis on electrophysiologic criteria. PMID- 19564583 TI - Optical coherence tomography is less sensitive than visual evoked potentials in optic neuritis. AB - OBJECTIVES: Determine the utility of optical coherence tomography (OCT) to detect clinical and subclinical remote optic neuritis (ON), its relationship to clinical characteristics of ON and visual function, and whether the retinal nerve fiber layer (RNFL) thickness functions as a surrogate marker of global disease severity. METHODS: Cross-sectional study of 65 subjects with at least 1 clinical ON episode at least 6 months prior. Measures included clinical characteristics, visual acuity (VA), contrast sensitivity (CS), OCT, and visual evoked potentials (VEP). RESULTS: Ninety-six clinically affected optic nerves were studied. The sensitivity of OCT RNFL after ON was 60%, decreasing further with mild onset and good recovery. VEP sensitivity was superior at 81% (p = 0.002). Subclinical ON in the unaffected eye was present in 32%. VEP identified 75% of all subclinically affected eyes, and OCT identified <20%. RNFL thickness demonstrated linear correlations with VA (r = 0.65) and CS (r = 0.72) but was unable to distinguish visual categories <20/50. RNFL was thinner with severe onset and disease recurrence but was unaffected by IV glucocorticoids. OCT measurements were not related to overall disability, ethnicity, sex, or age at onset. The greatest predictor for RNFL in the unaffected eye was the RNFL in the fellow affected eye. CONCLUSIONS: Visual evoked potentials (VEP) remains the preferred test for detecting clinical and subclinical optic neuritis. Optical coherence tomography (OCT) measures were unrelated to disability and demographic features predicting a worse prognosis in multiple sclerosis. OCT may provide complementary information to VEP in select cases, and remains a valuable research tool for studying optic nerve disease in populations. PMID- 19564584 TI - Long-term effects of pallidal deep brain stimulation in tardive dystonia. AB - OBJECTIVE: High-frequency stimulation of the globus pallidus internus (GPi) is a highly effective therapy in primary dystonia. Recent reports have also demonstrated almost immediate improvement of motor symptoms in patients with tardive dystonia after pallidal deep brain stimulation (DBS). Here, we show the long-term effect of continuous bilateral GPi DBS in tardive dystonia on motor function, quality of life (QoL), and mood. METHODS: Nine consecutive patients undergoing DBS for tardive dystonia were assessed during continuous DBS at 3 time points: 1 week, 3 to 6 months, and last follow-up at the mean of 41 (range 18-80) months after surgery using established and validated movement disorder and neuropsychological scales. Clinical assessment was performed by a neurologist not blinded to the stimulation settings. RESULTS: One week and 3 to 6 months after pallidal DBS, Burke-Fahn-Marsden Dystonia Rating Scale (BFMDRS) motor scores were ameliorated by 56.4 +/- 26.7% and 74.1 +/- 15.8%, BFMDRS disability scores by 62.5 +/- 21.0% and 88.9 +/- 10.3%, and Abnormal Involuntary Movement Scale (AIMS) scores by 52.3 +/- 24.1% and 69.5 +/- 27.6%, respectively. At last follow-up, this improvement compared with the presurgical assessment was maintained as reflected by a reduction of BFMDRS motor scores by 83.0 +/- 12.2%, BFMDRS disability scores by 67.7 +/- 28.0%, and AIMS scores by 78.7 +/- 19.9%. QoL improved significantly in physical components, and there was a significant improvement in affective state. Furthermore, cognitive functions remained unchanged compared with presurgical status in the long-term follow-up. No permanent adverse effects were observed. CONCLUSION: Pallidal deep brain stimulation is a safe and effective long-term treatment in patients with medically refractory tardive dystonia. PMID- 19564585 TI - Sudomotor, skin vasomotor, and cardiovascular reflexes in 3 clinical forms of Lewy body disease. AB - OBJECTIVE: To elucidate the differences among dementia with Lewy bodies (DLB), Parkinson disease with dementia (PDD), and Parkinson disease without dementia (PD), with respect to the involvement of the autonomic nervous system, we clinically investigated the cutaneous and cardiovascular autonomic functions in patients with Lewy body disease. METHODS: We studied 36 patients with Lewy body disorders, including 12 patients with DLB (age, 75.4 +/- 5.9 years), 12 patients with PDD (71.0 +/- 6.8 years), and 12 patients with PD (70.9 +/- 4.2 years), and 12 healthy control subjects (69.9 +/- 5.3 years). Sympathetic sweat response (SSwR) and skin vasomotor reflex (SkVR) on the palm were recorded to estimate the cutaneous sympathetic function, and the head-up tilt test was performed and coefficient of variation of R-R intervals (CV(R-R)) was studied to estimate the cardiovascular function. RESULTS: The patients with DLB, patients with PDD, and patients with PD showed severely reduced SSwR amplitudes, significantly lower than that in the controls. The mean SkVR amplitudes in the patients with DLB and patients with PDD were significantly lower than that in the controls, but not in the patients with PD. The mean decreases in the systolic blood pressure during the head-up tilt test in the patients with DLB and patients with PDD were less than that in the controls. The mean CV(R-R) value was significantly lower in the patients with DLB. CONCLUSION: Sudomotor function on the palm may be severely affected in Lewy body disorders, while skin vasomotor function and the cardiovascular system may be more severely affected in dementia with Lewy bodies and Parkinson disease with dementia than in Parkinson disease. PMID- 19564586 TI - Brain lesions are most often reversible in acute thrombotic thrombocytopenic purpura. AB - BACKGROUND: Thrombotic thrombocytopenic purpura (TTP) is a rare hematologic disorder that frequently presents with neurologic involvement. However, the yield and prognostic value of acute brain neuroimaging in patients with TTP has not been studied. Our aim was to evaluate brain imaging findings in consecutive patients with TTP and assess their impact on prognosis. METHODS: We retrospectively collected clinical, laboratory, and neuroradiologic information in 47 episodes of acute TTP studied with brain imaging at our medical center between 1997 and 2007. Head CT and brain MRI were evaluated independently by 2 investigators. We then performed statistical analysis to determine whether the presence of acute lesions on brain imaging was associated with worse functional outcome as assessed by the modified Rankin score upon discharge and long-term follow-up. RESULTS: Ten patients (25%) of those who had a head CT had acute changes, half of them indicating posterior reversible encephalopathy syndrome (PRES). Most cases studied with brain MRI had acute changes (82%). More than half of those had evidence of PRES (48%). Atypical variants of PRES were seen in 2 patients with isolated basal ganglia involvement. Acute ischemia and hemorrhage were uncommon. Most patients with acute changes on brain imaging recovered favorably, and radiologic lesions were not associated with worse functional outcome. CONCLUSIONS: Posterior reversible encephalopathy syndrome is the most common brain imaging abnormality in severe manifestations of thrombotic thrombocytopenic purpura. Large infarctions and hemorrhage are infrequent. Consequently, abnormal brain neuroimaging does not seem to impact patient outcome, and full neurologic recovery is possible even in comatose patients with extensive brain abnormalities on MRI. PMID- 19564587 TI - Ophthalmoplegia and ptosis: mitochondrial toxicity in patients receiving HIV therapy. PMID- 19564588 TI - Cerebellar ataxia after malaria. PMID- 19564589 TI - Bilateral cochlear enhancement in Cogan syndrome. PMID- 19564590 TI - Re: CDKL5 mutations in boys with severe encephalopathy and early-onset intractable epilepsy. PMID- 19564591 TI - Denture cream: an unusual source of excess zinc, leading to hypocupremia and neurologic disease. PMID- 19564592 TI - Re: CDKL5 mutations in boys with severe encephalopathy and early-onset intractable epilepsy. PMID- 19564593 TI - Typological labeling of early Holocene projectile points. PMID- 19564595 TI - Large and unexpected enrichment in stratospheric 16O13C18O and its meridional variation. AB - The stratospheric CO(2) oxygen isotope budget is thought to be governed primarily by the O((1)D)+CO(2) isotope exchange reaction. However, there is increasing evidence that other important physical processes may be occurring that standard isotopic tools have been unable to identify. Measuring the distribution of the exceedingly rare CO(2) isotopologue (16)O(13)C(18)O, in concert with (18)O and (17)O abundances, provides sensitivities to these additional processes and, thus, is a valuable test of current models. We identify a large and unexpected meridional variation in stratospheric (16)O(13)C(18)O, observed as proportions in the polar vortex that are higher than in any naturally derived CO(2) sample to date. We show, through photochemical experiments, that lower (16)O(13)C(18)O proportions observed in the midlatitudes are determined primarily by the O((1)D)+CO(2) isotope exchange reaction, which promotes a stochastic isotopologue distribution. In contrast, higher (16)O(13)C(18)O proportions in the polar vortex show correlations with long-lived stratospheric tracer and bulk isotope abundances opposite to those observed at midlatitudes and, thus, opposite to those easily explained by O((1)D)+CO(2). We believe the most plausible explanation for this meridional variation is either an unrecognized isotopic fractionation associated with the mesospheric photochemistry of CO(2) or temperature-dependent isotopic exchange on polar stratospheric clouds. Unraveling the ultimate source of stratospheric (16)O(13)C(18)O enrichments may impose additional isotopic constraints on biosphere-atmosphere carbon exchange, biosphere productivity, and their respective responses to climate change. PMID- 19564596 TI - Expected rate of fisheries-induced evolution is slow. AB - Commercial fisheries exert high mortalities on the stocks they exploit, and the consequent selection pressure leads to fisheries-induced evolution of growth rate, age and size at maturation, and reproductive output. Productivity and yields may decline as a result, but little is known about the rate at which such changes are likely to occur. Fisheries-induced evolution of exploited populations has recently become a subject of concern for policy makers, fisheries managers, and the general public, with prominent calls for mitigating management action. We make a general evolutionary impact assessment of fisheries by calculating the expected rate of fisheries-induced evolution and the consequent changes in yield. Rates of evolution are expected to be approximately 0.1-0.6% per year, and the consequent reductions in fisheries yield are <0.7% per year. These rates are at least a factor of 5 lower than published values based on experiments and analyses of population time series, and we explain why the published rates may be overestimates. Dealing with evolutionary effects of fishing is less urgent than reducing the direct detrimental effects of overfishing on exploited stocks and on their marine ecosystems. PMID- 19564597 TI - Syncytin-A knockout mice demonstrate the critical role in placentation of a fusogenic, endogenous retrovirus-derived, envelope gene. AB - In most mammalian species, a key process of placenta development is the fusion of trophoblast cells into a highly specialized, multinucleated syncytiotrophoblast layer, through which most of the maternofetal exchanges take place. Little is known about this process, despite the recent identification of 2 pairs of envelope genes of retroviral origin, independently acquired by the human (syncytin-1 and syncytin-2) and mouse (syncytin-A and syncytin-B) genomes, specifically expressed in the placenta, and with in vitro cell-cell fusion activity. By generating knockout mice, we show here that homozygous syncytin-A null mouse embryos die in utero between 11.5 and 13.5 days of gestation. Refined cellular and subcellular analyses of the syncytin-A-deficient placentae disclose specific disruption of the architecture of the syncytiotrophoblast-containing labyrinth, with the trophoblast cells failing to fuse into an interhemal syncytial layer. Lack of syncytin-A-mediated trophoblast cell fusion is associated with cell overexpansion at the expense of fetal blood vessel spaces and with apoptosis, adding to the observed maternofetal interface structural defects to provoke decreased vascularization, inhibition of placental transport, and fetal growth retardation, ultimately resulting in death of the embryo. These results demonstrate that syncytin-A is essential for trophoblast cell differentiation and syncytiotrophoblast morphogenesis during placenta development, and they provide evidence that genes captured from ancestral retroviruses have been pivotal in the acquisition of new, important functions in mammalian evolution. PMID- 19564598 TI - Secretory phospholipase A2-IID is an effector molecule of CD4+CD25+ regulatory T cells. AB - Suppression by natural CD4(+)CD25(+) regulatory T cells (Tregs) is one mechanism by which tolerance is maintained. However, the way in which Tregs mediate suppression is not well understood. Here, we show that secreted phospholipase A2 (sPLA2)-IID is selectively produced by Tregs. sPLA2-IID is a potent mediator of Treg function, because it strongly suppressed proliferation of CD4(+) and CD8(+) T cells in vitro and in vivo in a manner independent of its catalytic activity. Furthermore, sPLA2-IID promoted the differentiation of Tregs, presumably via attenuating signaling through the PI3K/Akt/mammalian target of rapamycin pathway. Importantly, administration of a sPLA2-IID-Fc fusion protein inhibited disease development in murine models of colitis and multiple sclerosis, suggesting that sPLA2-IID's immunosuppressive function might be exploited therapeutically. PMID- 19564599 TI - Biological consequences of earlier snowmelt from desert dust deposition in alpine landscapes. AB - Dust deposition to mountain snow cover, which has increased since the late 19(th) century, accelerates the rate of snowmelt by increasing the solar radiation absorbed by the snowpack. Snowmelt occurs earlier, but is decoupled from seasonal warming. Climate warming advances the timing of snowmelt and early season phenological events (e.g., the onset of greening and flowering); however, earlier snowmelt without warmer temperatures may have a different effect on phenology. Here, we report the results of a set of snowmelt manipulations in which radiation absorbing fabric and the addition and removal of dust from the surface of the snowpack advanced or delayed snowmelt in the alpine tundra. These changes in the timing of snowmelt were superimposed on a system where the timing of snowmelt varies with topography and has been affected by increased dust loading. At the community level, phenology exhibited a threshold response to the timing of snowmelt. Greening and flowering were delayed before seasonal warming, after which there was a linear relationship between the date of snowmelt and the timing of phenological events. Consequently, the effects of earlier snowmelt on phenology differed in relation to topography, which resulted in increasing synchronicity in phenology across the alpine landscape with increasingly earlier snowmelt. The consequences of earlier snowmelt from increased dust deposition differ from climate warming and include delayed phenology, leading to synchronized growth and flowering across the landscape and the opportunity for altered species interactions, landscape-scale gene flow via pollination, and nutrient cycling. PMID- 19564600 TI - A site-specific, multiplexed kinase activity assay using stable-isotope dilution and high-resolution mass spectrometry. AB - Most kinases are capable of recognizing and phosphorylating peptides containing short, linear sequence motifs. To measure the activation state of many kinases from the same cell lysate, we created a multiplexed, mass-spectrometry-based in vitro kinase assay. Ninety chemically synthesized peptides derived from well characterized peptide substrates and in vivo phosphorylation sites with either known or previously unidentified upstream kinases were reacted individually in a plate format with crude cell lysates and ATP. Phosphorylation rates were directly measured based on the addition of 90 same-sequence, site-specific phosphopeptides enriched in stable isotopes to act as ideal quantitative internal standards for analysis by liquid chromatography coupled to tandem mass spectrometry. This approach concurrently measured up to 90 site-specific peptide phosphorylation rates, reporting a diagnostic fingerprint for activated kinase pathways. We applied this unique kinome-activity profiling strategy in a variety of cellular settings, including mitogen stimulation, cell cycle, pharmacological inhibition of pathways, and to a panel of breast cancer cell lines. Finally, we identified the source of activity for a peptide (derived from a PI3K regulatory subunit) from our library. This peptide substrate demonstrated mitotic and tyrosine specific phosphorylation, which was confirmed to be a novel Src family kinase site in vivo. PMID- 19564601 TI - Invasive hybrid tiger salamander genotypes impact native amphibians. AB - Although the ecological consequences of species invasions are well studied, the ecological impacts of genetic introgression through hybridization are less understood. This is particularly true of the impacts of hybridization on "third party" community members not genetically involved in hybridization. We also know little about how direct interactions between hybrid and parental individuals influence fitness. Here, we examined the ecological effects of hybridization between the native, threatened California Tiger Salamander (Ambystoma californiense) and the introduced Barred Tiger Salamander (Ambystoma tigrinum mavortium). Native x introduced hybrids are widespread in California, where they are top predators in seasonal ponds. We examined the impacts of early generation hybrids (first 2 generations of parental crosses) and contemporary hybrids derived from ponds where hybrids have been under selection in the wild for 20 generations. We found that most classes of hybrid tiger salamander larvae dramatically reduced survival of 2 native community members, the Pacific Chorus Frog (Pseudacris regilla) and the California Newt (Taricha torosa). We also found that native A. californiense larvae were negatively impacted by the presence of hybrid larvae: Native survival and size at metamorphosis were reduced and time to metamorphosis was extended. We also observed a large influence of Mendelian dominance on size, metamorphic timing and predation rate of hybrid tiger salamanders. These results suggest that both genetic and ecological factors are likely to influence the dynamics of admixture, and that tiger salamander hybridization might constitute a threat to additional pond-breeding species of concern in the region. PMID- 19564602 TI - The private rejection of unfair offers and emotional commitment. AB - In a series of experiments, we demonstrate that certain players of an economic game reject unfair offers even when this behavior increases rather than decreases inequity. A substantial proportion (30-40%, compared with 60-70% in the standard ultimatum game) of those who responded rejected unfair offers even when rejection reduced only their own earnings to 0, while not affecting the earnings of the person who proposed the unfair split (in an impunity game). Furthermore, even when the responders were not able to communicate their anger to the proposers by rejecting unfair offers in a private impunity game, a similar rate of rejection was observed. The rejection of unfair offers that increases inequity cannot be explained by the social preference for inequity aversion or reciprocity; however, it does provide support for the model of emotion as a commitment device. In this view, emotions such as anger or moral disgust lead people to disregard the immediate consequences of their behavior, committing them to behave consistently to preserve integrity and maintain a reputation over time as someone who is reliably committed to this behavior. PMID- 19564603 TI - Quantitative analysis of dental microwear in hadrosaurid dinosaurs, and the implications for hypotheses of jaw mechanics and feeding. AB - Understanding the feeding mechanisms and diet of nonavian dinosaurs is fundamental to understanding the paleobiology of these taxa and their role in Mesozoic terrestrial ecosystems. Various methods, including biomechanical analysis and 3D computer modeling, have been used to generate detailed functional hypotheses, but in the absence of either direct observations of dinosaur feeding behavior, or close living functional analogues, testing these hypotheses is problematic. Microscopic scratches that form on teeth in vivo during feeding are known to record the relative motion of the tooth rows to each other during feeding and to capture evidence of tooth-food interactions. Analysis of this dental microwear provides a powerful tool for testing hypotheses of jaw mechanics, diet, and trophic niche; yet, quantitative analysis of microwear in dinosaurs has not been attempted. Here, we show that analysis of tooth microwear orientation provides direct evidence for the relative motions of jaws during feeding in hadrosaurid ornithopods, the dominant terrestrial herbivores of the Late Cretaceous. Statistical testing demonstrates that Edmontosaurus teeth preserve 4 distinct sets of scratches in different orientations. In terms of jaw mechanics, these data indicate an isognathic, near-vertical posterodorsal power stroke during feeding; near-vertical jaw opening; and propalinal movements in near anterior and near posterior directions. Our analysis supports the presence of a pleurokinetic hinge, and the straightness and parallelism of scratches indicate a tightly controlled occlusion. The dominance of scratched microwear fabrics suggests that Edmontosaurus was a grazer rather than a browser. PMID- 19564604 TI - Requirement of central ghrelin signaling for alcohol reward. AB - The stomach-derived hormone ghrelin interacts with key CNS circuits regulating energy balance and body weight. Here we provide evidence that the central ghrelin signaling system is required for alcohol reward. Central ghrelin administration (to brain ventricles or to tegmental areas involved in reward) increased alcohol intake in a 2-bottle (alcohol/water) free choice limited access paradigm in mice. By contrast, central or peripheral administration of ghrelin receptor (GHS-R1A) antagonists suppressed alcohol intake in this model. Alcohol-induced locomotor stimulation, accumbal dopamine release and conditioned place preference were abolished in models of suppressed central ghrelin signaling: GHS-R1A knockout mice and mice treated with 2 different GHS-R1A antagonists. Thus, central ghrelin signaling, via GHS-R1A, not only stimulates the reward system, but is also required for stimulation of that system by alcohol. Our data suggest that central ghrelin signaling constitutes a potential target for treatment of alcohol-related disorders. PMID- 19564605 TI - Cognitive fitness of cost-efficient brain functional networks. AB - The human brain's capacity for cognitive function is thought to depend on coordinated activity in sparsely connected, complex networks organized over many scales of space and time. Recent work has demonstrated that human brain networks constructed from neuroimaging data have economical small-world properties that confer high efficiency of information processing at relatively low connection cost. However, it has been unclear how the architecture of complex brain networks functioning at different frequencies can be related to behavioral performance on cognitive tasks. Here, we show that impaired accuracy of working memory could be related to suboptimal cost efficiency of brain functional networks operating in the classical beta frequency band, 15-30 Hz. We analyzed brain functional networks derived from magnetoencephalography data recorded during working-memory task performance in 29 healthy volunteers and 28 people with schizophrenia. Networks functioning at higher frequencies had greater global cost efficiency than low-frequency networks in both groups. Superior task performance was positively correlated with global cost efficiency of the beta-band network and specifically with cost efficiency of nodes in left lateral parietal and frontal areas. These results are consistent with biophysical models highlighting the importance of beta-band oscillations for long-distance functional connections in brain networks and with pathophysiological models of schizophrenia as a dysconnection syndrome. More generally, they echo the saying that "less is more": The information processing performance of a network can be enhanced by a sparse or low-cost configuration with disproportionately high efficiency. PMID- 19564606 TI - Kinetic theories of dynamics and persistent caging in a one-dimensional lattice gas. AB - The one-dimensional, single-occupancy lattice gas exhibits highly cooperative particle motion and provides an interesting challenge for theoretical methods designed to describe caging in liquids. We employ this model in an effort to gain insight into caging phenomena in more realistic models of liquids, using a diagrammatic kinetic theory of density fluctuations to develop a series of approximations to the kinetic equations for the van Hove self-correlation function. The approximations are formulated in terms of the irreducible memory function, and we assess their efficacy by comparing their solutions with computer simulation results and the well-known subdiffusive behavior of a tagged particle at long times. The first approximation, a mode coupling theory, factorizes the 4 point propagators that contribute to the irreducible memory function into products of independent single-particle propagators. This approximation fails to capture the subdiffusive behavior of a tagged particle at long times. Analysis of the mode coupling approximation in terms of the diagrammatic kinetic theory leads to the development of two additional approximations that can be viewed as diagrammatic extensions or modifications of mode coupling theory. The first, denoted MC1, captures the long-time subdiffusive behavior of a tagged particle. The second, denoted MC2, captures the subdiffusive behavior of a tagged particle and also yields the correct amplitude of its mean square displacement at long times. Numerical and asymptotic solutions of the approximate kinetic equations share many qualitative and quantitative features with simulation results at all timescales. PMID- 19564607 TI - Bacteria capture iron from heme by keeping tetrapyrrol skeleton intact. AB - Because heme is a major iron-containing molecule in vertebrates, the ability to use heme-bound iron is a determining factor in successful infection by bacterial pathogens. Until today, all known enzymes performing iron extraction from heme did so through the rupture of the tetrapyrrol skeleton. Here, we identified 2 Escherichia coli paralogs, YfeX and EfeB, without any previously known physiological functions. YfeX and EfeB promote iron extraction from heme preserving the tetrapyrrol ring intact. This novel enzymatic reaction corresponds to the deferrochelation of the heme. YfeX and EfeB are the sole proteins able to provide iron from exogenous heme sources to E. coli. YfeX is located in the cytoplasm. EfeB is periplasmic and enables iron extraction from heme in the periplasm and iron uptake in the absence of any heme permease. YfeX and EfeB are widespread and highly conserved in bacteria. We propose that their physiological function is to retrieve iron from heme. PMID- 19564608 TI - Control of retinal isomerization in bacteriorhodopsin in the high-intensity regime. AB - A learning algorithm was used to manipulate optical pulse shapes and optimize retinal isomerization in bacteriorhodopsin, for excitation levels up to 1.8 x 10(16) photons per square centimeter. Below 1/3 the maximum excitation level, the yield was not sensitive to pulse shape. Above this level the learning algorithm found that a Fourier-transform-limited (TL) pulse maximized the 13-cis population. For this optimal pulse the yield increases linearly with intensity well beyond the saturation of the first excited state. To understand these results we performed systematic searches varying the chirp and energy of the pump pulses while monitoring the isomerization yield. The results are interpreted including the influence of 1-photon and multiphoton transitions. The population dynamics in each intermediate conformation and the final branching ratio between the all-trans and 13-cis isomers are modified by changes in the pulse energy and duration. PMID- 19564609 TI - Major flowering time gene, flowering locus C, regulates seed germination in Arabidopsis thaliana. AB - Flowering locus C (FLC) is a major regulator of flowering responses to seasonal environmental factors. Here, we document that FLC also regulates another major life-history transition-seed germination, and that natural variation at the FLC locus and in FLC expression is associated with natural variation in temperature dependent germination. FLC-mediated germination acts through additional genes in the flowering pathway (FT, SOC1, and AP1) before involving the abscisic acid catabolic pathway (via CYP707A2) and gibberellins biosynthetic pathway (via GA20ox1) in seeds. Also, FLC regulation of germination is largely maternally controlled, with FLC peaking and FT, SOC1, and AP1 levels declining at late stages of seed maturation. High FLC expression during seed maturation is associated with altered expression of hormonal genes (CYP707A2 and GA20ox1) in germinating seeds, indicating that gene expression before the physiological independence of seeds can influence gene expression well after any physical connection between maternal plants and seeds exists. The major role of FLC in temperature-dependent germination documented here reveals a much broader adaptive significance of natural variation in FLC. Therefore, pleiotropy between these major life stages likely influences patterns of natural selection on this important gene, making FLC a promising case for examining how pleiotropy influences adaptive evolution. PMID- 19564610 TI - Linking social norms to efficient conservation investment in payments for ecosystem services. AB - An increasing amount of investment has been devoted to protecting and restoring ecosystem services worldwide. The efficiency of conservation investments, including payments for ecosystem services (PES), has been found to be affected by biological, political, economic, demographic, and social factors, but little is known about the effects of social norms at the neighborhood level. As a first attempt to quantify the effects of social norms, we studied the effects of a series of possible factors on people's intentions of maintaining forest on their Grain-to-Green Program (GTGP) land plots if the program ends. GTGP is one of the world's largest PES programs and plays an important role in global conservation efforts. Our study was conducted in China's Wolong Nature Reserve, home to the world-famous endangered giant pandas and >4,500 farmers. We found that, in addition to conservation payment amounts and program duration, social norms at the neighborhood level had significant impacts on program re-enrollment, suggesting that social norms can be used to leverage participation to enhance the sustainability of conservation benefits from PES programs. Moreover, our results demonstrate that economic and demographic trends also have profound implications for sustainable conservation. Thus, social norms should be incorporated with economic and demographic trends for efficient conservation investments. PMID- 19564611 TI - Toll-like receptor signaling controls reactivation of KSHV from latency. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent of Kaposi's sarcoma, primary effusion lymphoma (PEL), and multicentric Castleman's disease. Like other herpesviruses, KSHV establishes life-long latency in the human host with intermittent periods of reactivation. Physiological triggers of herpesviral reactivation are poorly defined. Toll-like receptors (TLRs) recognize pathogens and are vital for the host innate immune response. We screened multiple TLR agonists for their ability to initiate KSHV replication in latently infected PEL. Agonists specific for TLR7/8 reactivated latent KSHV and induced viral lytic gene transcription and replication. Furthermore, vesicular stomatitis virus (VSV), a bonafide physiological activator of TLR7/8, also reactivated KSHV from latency. This demonstrates that secondary pathogen infection of latently infected cells can reactivate KSHV. Human herpesviruses establish life-long latency in the host, and it is plausible that a latently infected cell will encounter multiple pathogens during its lifetime and that these encounters lead to episodic reactivation. Our findings have broad implications for physiological triggers of latent viral infections, such as herpesviral reactivation and persistence in the host. PMID- 19564612 TI - Evidence for a memory threshold in second-choice recognition memory responses. AB - A fundamental question in the study of cognition is whether memory strength varies continuously or whether memories sometimes fall below a threshold and fail completely. Previous studies examining this question have relied exclusively on 1 method--receiver operating characteristics--so in the current study, we addressed this issue by using a completely different approach. We tested memory for single items and for arbitrary associations (e.g., memory for random word pairs) by using a 4-alternative forced-choice test in which subjects either made a single choice or a first and a second choice. In item recognition, single- and second choice scores were directly related, as expected if a continuous strength signal supported performance. In contrast, in associative recognition, single- and second-choice scores were found to be unrelated, as predicted by high-threshold theories. However, when the word pairs were encoded as single compound words rather than arbitrary associations, associative recognition appeared to rely more on a continuous strength process. The results support memory models that include both a continuous familiarity process and a threshold recollection process. PMID- 19564614 TI - Cholinergic filtering in the recurrent excitatory microcircuit of cortical layer 4. AB - Neocortical acetylcholine (ACH) release is known to enhance signal processing by increasing the amplitude and signal-to-noise ratio (SNR) of sensory responses. It is widely accepted that the larger sensory responses are caused by a persistent increase in the excitability of all cortical excitatory neurons. Here, contrary to this concept, we show that ACH persistently inhibits layer 4 (L4) spiny neurons, the main targets of thalamocortical inputs. Using whole-cell recordings in slices of rat primary somatosensory cortex, we demonstrate that this inhibition is specific to L4 and contrasts with the ACH-induced persistent excitation of pyramidal cells in L2/3 and L5. We find that this inhibition is induced by postsynaptic M(4)-muscarinic ACH receptors and is mediated by the opening of inwardly rectifying potassium (K(ir)) channels. Pair recordings of L4 spiny neurons show that ACH reduces synaptic release in the L4 recurrent microcircuit. We conclude that ACH has a differential layer-specific effect that results in a filtering of weak sensory inputs in the L4 recurrent excitatory microcircuit and a subsequent amplification of relevant inputs in L2/3 and L5 excitatory microcircuits. This layer-specific effect may contribute to improve cortical SNR. PMID- 19564615 TI - Inhibition of serine/threonine phosphatase PP2A enhances cancer chemotherapy by blocking DNA damage induced defense mechanisms. AB - A variety of mechanisms maintain the integrity of the genome in the face of cell stress. Cancer cell response to chemotherapeutic and radiation-induced DNA damage is mediated by multiple defense mechanisms including polo-like kinase 1 (Plk-1), protein kinase B (Akt-1), and/or p53 pathways leading to either apoptosis or cell cycle arrest. Subsequently, a subpopulation of arrested viable cancer cells may remain and recur despite aggressive and repetitive therapy. Here, we show that modulation (activation of Akt-1 and Plk-1 and repression of p53) of these pathways simultaneously results in paradoxical enhancement of the effectiveness of cytotoxic chemotherapy. We demonstrate that a small molecule inhibitor, LB 1.2, of protein phosphatase 2A (PP2A) activates Plk-1 and Akt-1 and decreases p53 abundance in tumor cells. Combined with temozolomide (TMZ; a DNA-methylating chemotherapeutic drug), LB-1.2 causes complete regression of glioblastoma multiforme (GBM) xenografts without recurrence in 50% of animals (up to 28 weeks) and complete inhibition of growth of neuroblastoma (NB) xenografts. Treatment with either drug alone results in only short-term inhibition/regression with all xenografts resuming rapid growth. Combined with another widely used anticancer drug, Doxorubicin (DOX, a DNA intercalating agent), LB-1.2 also causes marked GBM xenograft regression, whereas DOX alone only slows growth. Inhibition of PP2A by LB-1.2 blocks cell-cycle arrest and increases progression of cell cycle in the presence of TMZ or DOX. Pharmacologic inhibition of PP2A may be a general method for enhancing the effectiveness of cancer treatments that damage DNA or disrupt components of cell replication. PMID- 19564616 TI - High-resolution profiling and discovery of planarian small RNAs. AB - Freshwater planarian flatworms possess uncanny regenerative capacities mediated by abundant and collectively totipotent adult stem cells. Key functions of these cells during regeneration and tissue homeostasis have been shown to depend on PIWI, a molecule required for Piwi-interacting RNA (piRNA) expression in planarians. Nevertheless, the full complement of piRNAs and microRNAs (miRNAs) in this organism has yet to be defined. Here we report on the large-scale cloning and sequencing of small RNAs from the planarian Schmidtea mediterranea, yielding altogether millions of sequenced, unique small RNAs. We show that piRNAs are in part organized in genomic clusters and that they share characteristic features with mammalian and fly piRNAs. We further identify 61 novel miRNA genes and thus double the number of known planarian miRNAs. Sequencing, as well as quantitative PCR of small RNAs, uncovered 10 miRNAs enriched in planarian stem cells. These miRNAs are down-regulated in animals in which stem cells have been abrogated by irradiation, and thus constitute miRNAs likely associated with specific stem-cell functions. Altogether, we present the first comprehensive small RNA analysis in animals belonging to the third animal superphylum, the Lophotrochozoa, and single out a number of miRNAs that may function in regeneration. Several of these miRNAs are deeply conserved in animals. PMID- 19564617 TI - TRPV1-expressing primary afferents generate behavioral responses to pruritogens via multiple mechanisms. AB - The mechanisms that generate itch are poorly understood at both the molecular and cellular levels despite its clinical importance. To explore the peripheral neuronal mechanisms underlying itch, we assessed the behavioral responses (scratching) produced by s.c. injection of various pruritogens in PLCbeta3- or TRPV1-deficient mice. We provide evidence that at least 3 different molecular pathways contribute to the transduction of itch responses to different pruritogens: 1) histamine requires the function of both PLCbeta3 and the TRPV1 channel; 2) serotonin, or a selective agonist, alpha-methyl-serotonin (alpha-Me-5 HT), requires the presence of PLCbeta3 but not TRPV1, and 3) endothelin-1 (ET-1) does not require either PLCbeta3 or TRPV1. To determine whether the activity of these molecules is represented in a particular subpopulation of sensory neurons, we examined the behavioral consequences of selectively eliminating 2 nonoverlapping subsets of nociceptors. The genetic ablation of MrgprD(+) neurons that represent approximately 90% of cutaneous nonpeptidergic neurons did not affect the scratching responses to a number of pruritogens. In contrast, chemical ablation of the central branch of TRPV1(+) nociceptors led to a significant behavioral deficit for pruritogens, including alpha-Me-5-HT and ET-1, that is, the TRPV1-expressing nociceptor was required, whether or not TRPV1 itself was essential. Thus, TRPV1 neurons are equipped with multiple signaling mechanisms that respond to different pruritogens. Some of these require TRPV1 function; others use alternate signal transduction pathways. PMID- 19564618 TI - DNA polymerase zeta cooperates with polymerases kappa and iota in translesion DNA synthesis across pyrimidine photodimers in cells from XPV patients. AB - Human cells tolerate UV-induced cyclobutane pyrimidine dimers (CPD) by translesion DNA synthesis (TLS), carried out by DNA polymerase eta, the POLH gene product. A deficiency in DNA polymerase eta due to germ-line mutations in POLH causes the hereditary disease xeroderma pigmentosum variant (XPV), which is characterized by sunlight sensitivity and extreme predisposition to sunlight induced skin cancer. XPV cells are UV hypermutable due to the activity of mutagenic TLS across CPD, which explains the cancer predisposition of the patients. However, the identity of the backup polymerase that carries out this mutagenic TLS was unclear. Here, we show that DNA polymerase zeta cooperates with DNA polymerases kappa and iota to carry out error-prone TLS across a TT CPD. Moreover, DNA polymerases zeta and kappa, but not iota, protect XPV cells against UV cytotoxicity, independently of nucleotide excision repair. This presents an extreme example of benefit-risk balance in the activity of TLS polymerases, which provide protection against UV cytotoxicity at the cost of increased mutagenic load. PMID- 19564619 TI - High-precision optical measurements of 13C/12C isotope ratios in organic compounds at natural abundance. AB - A continuous-flow cavity ring-down spectroscopy (CRDS) system integrating a chromatographic separation technique, a catalytic combustor, and an isotopic (13)C/(12)C optical analyzer is described for the isotopic analysis of a mixture of organic compounds. A demonstration of its potential is made for the geochemically important class of short-chain hydrocarbons. The system proved to be linear over a 3-fold injection volume dynamic range with an average precision of 0.95 per thousand and 0.67 per thousand for ethane and propane, respectively. The calibrated accuracy for methane, ethane, and propane is within 3 per thousand of the values determined using isotope ratio mass spectrometry (IRMS), which is the current method of choice for compound-specific isotope analysis. With anticipated improvements, the low-cost, portable, and easy-to-use CRDS-based instrumental setup is poised to evolve into a credible challenge to the high-cost and complex IRMS-based technique. PMID- 19564620 TI - Ultrasonication-dependent production and breakdown lead to minimum-sized amyloid fibrils. AB - Because of the insolubility and polymeric properties of amyloid fibrils, techniques used conventionally to analyze protein structure and dynamics have often been hampered. Ultrasonication can induce the monomeric solution of amyloidogenic proteins to form amyloid fibrils. However, ultrasonication can break down preformed fibrils into shorter fibrils. Here, combining these 2 opposing effects on beta(2)-microglobulin (beta2-m), a protein responsible for dialysis-related amyloidosis, we present that ultrasonication pulses are useful for preparing monodispersed amyloid fibrils of minimal size with an average molecular weight of approximately 1,660,000 (140-mer). The production of minimal and monodispersed fibrils is achieved by the free energy minimum under competition between fibril production and breakdown. The small homogeneous fibrils will be of use for characterizing the structure and dynamics of amyloid fibrils, advancing molecular understanding of amyloidosis. PMID- 19564621 TI - Morphological record of oxygenic photosynthesis in conical stromatolites. AB - Conical stromatolites are thought to be robust indicators of the presence of photosynthetic and phototactic microbes in aquatic environments as early as 3.5 billion years ago. However, phototaxis alone cannot explain the ubiquity of disrupted, curled, and contorted laminae in the crests of many Mesoproterozoic, Paleoproterozoic, and some Archean conical stromatolites. Here, we demonstrate that cyanobacterial production of oxygen in the tips of modern conical aggregates creates contorted laminae and submillimeter-to-millimeter-scale enmeshed bubbles. Similarly sized fossil bubbles and contorted laminae may be present only in the crestal zones of some conical stromatolites 2.7 billion years old or younger. This implies not only that cyanobacteria built Proterozoic conical stromatolites but also that fossil bubbles may constrain the timing of the evolution of oxygenic photosynthesis. PMID- 19564622 TI - Pharmacokinetics and tumor dynamics of the nanoparticle IT-101 from PET imaging and tumor histological measurements. AB - IT-101, a cyclodextrin polymer-based nanoparticle containing camptothecin, is in clinical development for the treatment of cancer. Multiorgan pharmacokinetics and accumulation in tumor tissue of IT-101 is investigated by using PET. IT-101 is modified through the attachment of a 1,4,7,10-tetraazacyclododecane-1,4,7-Tris acetic acid ligand to bind (64)Cu(2+). This modification does not affect the particle size and minimally affects the surface charge of the resulting nanoparticles. PET data from (64)Cu-labeled IT-101 are used to quantify the in vivo biodistribution in mice bearing Neuro2A s.c. tumors. The (64)Cu-labeled IT 101 displays a biphasic plasma elimination. Approximately 8% of the injected dose is rapidly cleared as a low-molecular-weight fraction through the kidneys. The remaining material circulates in plasma with a terminal half-life of 13.3 h. Steadily increasing concentrations, up to 11% injected dose per cm(3), are observed in the tumor over 24 h, higher than any other tissue at that time. A 3 compartment model is used to determine vascular permeability and nanoparticle retention in tumors, and is able to accurately represent the experimental data. The calculated tumor vascular permeability indicates that the majority of nanoparticles stay intact in circulation and do not disassemble into individual polymer strands. A key assumption to modeling the tumor dynamics is that there is a "sink" for the nanoparticles within the tumor. Histological measurements using confocal microscopy show that IT-101 localizes within tumor cells and provides the sink in the tumor for the nanoparticles. PMID- 19564623 TI - Site-specific regulation of cell cycle and DNA repair in post-mitotic GABA cells in schizophrenic versus bipolars. AB - GABA cell dysfunction in both schizophrenia (SZ) and bipolar disorder (BD) involves decreased GAD(67) expression, although this change involves fundamentally different networks of genes in the 2 disorders. One gene that is common to these 2 networks is cyclin D2, a key component of cell cycle regulation that shows increased expression in SZ, but decreased expression in BD. Because of the importance of cell cycle regulation in maintaining functional differentiation and DNA repair, the current study has examined the genes involved in the G(1) and G(2) checkpoints to generate new hypotheses regarding the regulation of the GABA cell phenotype in the hippocampus of SZ and BD. The results have demonstrated significant changes in cell cycle regulation in both SZ and BD and these changes include the transcriptional complex (TC) that controls the expression of E2F/DP-1 target genes critical for progression to G(2)/M. The methyl-CpG binding domain protein (MBD4) that is pivotal for DNA repair, is significantly up-regulated in the stratum oriens (SO) of CA3/2 and CA1 in SZs and BDs. However, other genes associated with the TC, and the G(1) and G(2) checkpoints, show complex changes in expression in the SO of CA3/2 and CA1 of both SZs and BDS. Overall, the patterns of expression observed have suggested that the regulation of functional differentiation and/or genomic integrity of hippocampal GABA cells varies according to diagnosis and their location within the trisynaptic pathway. PMID- 19564624 TI - A "latent niche" mechanism for tumor initiation. AB - Stem cells, their niches, and their relationship to cancer are under intense investigation. Because tumors and metastases acquire self-renewing capacity, mechanisms for their establishment may involve cell-cell interactions similar to those between stem cells and stem cell niches. On the basis of our studies in Caenorhabditis elegans, we introduce the concept of a "latent niche" as a differentiated cell type that does not normally contact stem cells nor act as a niche but that can, under certain conditions, promote the ectopic self-renewal, proliferation, or survival of competent cells that it inappropriately contacts. Here, we show that ectopic germ-line stem cell proliferation in C. elegans is driven by a latent niche mechanism and that the molecular basis for this mechanism is inappropriate Notch activation. Furthermore, we show that continuous Notch signaling is required to maintain ectopic germ-line proliferation. We highlight the latent niche concept by distinguishing it from a normal stem cell niche, a premetastatic niche and an ectopic niche. One of the important distinguishing features of this mechanism for tumor initiation is that it could operate in the absence of genetic changes to the tumor cell or the tumor promoting cell. We propose that a latent niche mechanism may underlie tumorigenesis and metastasis in humans. PMID- 19564625 TI - Racial and geographic disparities in the use of parenteral nutrition among inflammatory bowel disease inpatients diagnosed with malnutrition in the United States. AB - BACKGROUND: Racial disparities have been described in the use of a diverse spectrum of surgical procedures. The objectives of this study are to determine whether disparities also exist for the use of parenteral nutrition (PN) in inflammatory bowel disease (IBD). METHODS: The U.S. Nationwide Inpatient Sample between 1998 and 2003 is analyzed to determine PN use among IBD inpatients diagnosed with protein-calorie malnutrition and assess whether use patterns differ by race and geographical region. RESULTS: The proportion of African American IBD admissions with protein-calorie malnutrition who receive PN is significantly lower than that in whites (19.9% vs 28.1%, P = .001), whereas there is no difference between Hispanics and non-Hispanic whites. After adjustment for gender, comorbidity, health insurance status, geographic region, and median neighborhood income, African Americans remain less likely than whites to receive PN (odds ratio [OR] 0.67; 95% confidence interval [CI], 0.50-0.89), whereas the difference between Hispanics and non-Hispanic whites is marginally significant (OR 0.65; 95% CI, 0.41-1.04). PN use varies geographically, with highest rates in the Northeast (44.3%) and lowest in the Midwest (17.3%). Uninsured patients are less than half as likely to receive PN as those with insurance (OR 0.46; 95% CI, 0.31-0.69). Compared with whites, Hispanics experience a longer time interval between admission and initiation of PN (3.5 vs 4.8 days, P = .02) and have higher rates of catheter-related complications (5.1% vs 12.2%, P = .04). CONCLUSIONS: Among IBD inpatients with clinically diagnosable malnutrition, PN use is lower among African Americans compared with whites. The underlying mechanisms of these racial variations merit further investigation. PMID- 19564626 TI - Effects of intravenous lipid emulsions in postoperative patients. PMID- 19564627 TI - Hyperglycemic events in non-intensive care unit patients receiving parenteral nutrition. AB - BACKGROUND: Evidence supports the benefits of tight glycemic control in many patient populations. There is no consensus on appropriate targets for blood glucose (BG) values in patients receiving parenteral nutrition (PN). Characterization of the frequency of BG abnormalities is necessary to identify effective strategies to improve glycemic control in this patient population. METHODS: Data were retrospectively collected over a 2-month period from 50 non intensive care unit (ICU) patients who received PN. Frequencies of abnormal BG (defined as BG outside the range of 2 criteria: 80-200 mg/dL and 100-150 mg/dL) were determined. An event of hyperglycemia was defined as the 48-hour period following a BG value outside of 80-200 mg/dL. Each event was evaluated for resolution within 48 hours of the triggering BG value. RESULTS: Hyperglycemia (at least 1 BG value >200 mg/dL) occurred in 22 patients (44%). Of the 1738 BG values measured, 8.7% were >200 mg/dL, resulting in 1.4 events of hyperglycemia per patient. The average blood glucose value for the population was 140 mg/dL. The frequency of hyperglycemia and hypoglycemia increased substantially, with only 1 patient having a PN course with normoglycemia using the 100-150 mg/dL criterion. CONCLUSION: The frequency of hyperglycemia in non-ICU PN patients is high according to either evaluation criterion. A method is described for using events to characterize hyperglycemia, which may be more useful than traditional methods in clinical decision making and identification of need for process improvements. These data suggest the need to develop better methods for BG control in non-ICU PN patients. PMID- 19564628 TI - Posttraumatic stress symptoms in children and adolescents referred for child welfare investigation. A national sample of in-home and out-of-home care. AB - This study examines the prevalence and correlates of heightened posttraumatic stress (PTS) symptoms in a nationally representative sample of 1,848 children and adolescents (ages 8-14) who were referred to child welfare for investigation of abuse or neglect based on the National Survey of Child and Adolescent Well-Being. The severity of current PTS symptoms was assessed using the PTS subscale of the Trauma Symptom Checklist for Children, a standardized child-report scale evaluating common symptoms associated with trauma. The overall prevalence of clinically significant PTS symptoms was 11.7% (overall mean T score = 49.5). The prevalence was higher for cases that were placed in out-of-home care (19.2%) than those maintained at home (10.7%). Multivariate hierarchical regression identified four contributors to heightened PTS symptoms: younger child age, abuse by a nonbiological parent, violence in the home, and child depression. The authors discuss the modest but still lower than expected prevalence of self-reported, clinically significant PTS symptoms and the variables associated with greater risk for heightened PTS symptoms found among cases referred to child welfare services. PMID- 19564629 TI - The persistent legacy of the 1918 influenza virus. PMID- 19564630 TI - Spread of a novel influenza A (H1N1) virus via global airline transportation. PMID- 19564631 TI - Pneumonia and respiratory failure from swine-origin influenza A (H1N1) in Mexico. AB - BACKGROUND: In late March 2009, an outbreak of a respiratory illness later proved to be caused by novel swine-origin influenza A (H1N1) virus (S-OIV) was identified in Mexico. We describe the clinical and epidemiologic characteristics of persons hospitalized for pneumonia at the national tertiary hospital for respiratory illnesses in Mexico City who had laboratory-confirmed S-OIV infection, also known as swine flu. METHODS: We used retrospective medical chart reviews to collect data on the hospitalized patients. S-OIV infection was confirmed in specimens with the use of a real-time reverse-transcriptase polymerase-chain-reaction assay. RESULTS: From March 24 through April 24, 2009, a total of 18 cases of pneumonia and confirmed S-OIV infection were identified among 98 patients hospitalized for acute respiratory illness at the National Institute of Respiratory Diseases in Mexico City. More than half of the 18 case patients were between 13 and 47 years of age, and only 8 had preexisting medical conditions. For 16 of the 18 patients, this was the first hospitalization for their illness; the other 2 patients were referred from other hospitals. All patients had fever, cough, dyspnea or respiratory distress, increased serum lactate dehydrogenase levels, and bilateral patchy pneumonia. Other common findings were an increased creatine kinase level (in 62% of patients) and lymphopenia (in 61%). Twelve patients required mechanical ventilation, and seven died. Within 7 days after contact with the initial case patients, a mild or moderate influenza-like illness developed in 22 health care workers; they were treated with oseltamivir, and none were hospitalized. CONCLUSIONS: S-OIV infection can cause severe illness, the acute respiratory distress syndrome, and death in previously healthy persons who are young to middle-aged. None of the secondary infections among health care workers were severe. PMID- 19564632 TI - Historical perspective--Emergence of influenza A (H1N1) viruses. PMID- 19564633 TI - Severe respiratory disease concurrent with the circulation of H1N1 influenza. AB - BACKGROUND: In the spring of 2009, an outbreak of severe pneumonia was reported in conjunction with the concurrent isolation of a novel swine-origin influenza A (H1N1) virus (S-OIV), widely known as swine flu, in Mexico. Influenza A (H1N1) subtype viruses have rarely predominated since the 1957 pandemic. The analysis of epidemic pneumonia in the absence of routine diagnostic tests can provide information about risk factors for severe disease from this virus and prospects for its control. METHODS: From March 24 to April 29, 2009, a total of 2155 cases of severe pneumonia, involving 821 hospitalizations and 100 deaths, were reported to the Mexican Ministry of Health. During this period, of the 8817 nasopharyngeal specimens that were submitted to the National Epidemiological Reference Laboratory, 2582 were positive for S-OIV. We compared the age distribution of patients who were reported to have severe pneumonia with that during recent influenza epidemics to document an age shift in rates of death and illness. RESULTS: During the study period, 87% of deaths and 71% of cases of severe pneumonia involved patients between the ages of 5 and 59 years, as compared with average rates of 17% and 32%, respectively, in that age group during the referent periods. Features of this epidemic were similar to those of past influenza pandemics in that circulation of the new influenza virus was associated with an off-season wave of disease affecting a younger population. CONCLUSIONS: During the early phase of this influenza pandemic, there was a sudden increase in the rate of severe pneumonia and a shift in the age distribution of patients with such illness, which was reminiscent of past pandemics and suggested relative protection for persons who were exposed to H1N1 strains during childhood before the 1957 pandemic. If resources or vaccine supplies are limited, these findings suggest a rationale for focusing prevention efforts on younger populations. PMID- 19564634 TI - Rapid-test sensitivity for novel swine-origin influenza A (H1N1) virus in humans. PMID- 19564635 TI - Definitive proof for direct reprogramming of hematopoietic cells to pluripotency. AB - Generation of induced pluripotent stem cells (iPSCs) generally uses fibroblastic cells, but other cell sources may prove useful in both research and clinical settings. Although proof of cellular origin requires genetic-marker identification in both target cells and established iPSCs, somatic cells other than mature lymphocytes mostly lack such markers. Here we show definitive proof of direct reprogramming of murine hematopoietic cells with no rearranged genes. Using iPSC factor transduction, we successfully derived iPSCs from bone marrow progenitor cells obtained from a mouse whose hematopoiesis was reconstituted from a single congenic hematopoietic stem cell. Established clones were demonstrated to be genetically identical to the transplanted single hematopoietic stem cell, thus proving their cellular origin. These hematopoietic cell-derived iPSCs showed typical characteristics of iPSCs, including the ability to contribute to chimerism in mice. These results will prompt further use of hematopoietic cells for iPSC generation while enabling definitive studies to test how cellular sources influence characteristics of descendant iPSCs. PMID- 19564638 TI - Phosphorylation of endothelial nitric oxide synthase by atypical PKC zeta contributes to angiopoietin-1-dependent inhibition of VEGF-induced endothelial permeability in vitro. AB - Vascular endothelial growth factor (VEGF) is a potent angiogenic cytokine that also increases vascular permeability. Nitric oxide (NO) released from endothelial cells, after activation of endothelial NO synthase (eNOS), contributes to proangiogenic and permeability effects of VEGF. Angiopoietin-1 (Ang-1), via Tie2 receptors, shares many of the proangiogenic properties of VEGF on endothelial cells. However, in contrast to VEGF, Ang-1 protects blood vessels from increased plasma leakage, which contributes to their stabilization. Because eNOS-derived NO is central to increased permeability in response to VEGF, we investigated whether Ang-1 interferes with VEGF signaling to eNOS. We demonstrate that Ang-1 stimulation of endothelial cells inhibits VEGF-induced NO release and transendothelial permeability. In contrast to VEGF stimulation, Ang-1 causes a marked protein kinase C (PKC)-dependent increase in phosphorylation of eNOS on the inhibitory Thr(497). Furthermore, using pharmacologic inhibitors, overexpression studies, and small interfering RNA-mediated gene silencing, we demonstrate that atypical PKC zeta is responsible for phosphorylation of eNOS on Thr(497) in response to Ang-1. In addition, PKC zeta knockdown abrogates the capacity of Ang-1 to inhibit VEGF-induced NO release and endothelial permeability. Thus, inhibition of NO production by Ang-1, via phosphorylation of eNOS on Thr(497) by PKC zeta, is responsible, at least in part, for inhibition of VEGF-stimulated endothelial permeability by Ang-1. PMID- 19564637 TI - Analysis of the ten-eleven translocation 2 (TET2) gene in familial myeloproliferative neoplasms. AB - The JAK2(V617F) mutation does not elucidate the phenotypic variability observed in myeloproliferative neoplasm (MPN) families. A putative tumor suppressor gene, TET2, was recently implicated in MPN and myelodysplastic syndromes through the identification of acquired mutations affecting hematopoietic stem cells. The present study analyzed the TET2 gene in 61 MPN cases from 42 families. Fifteen distinct mutations were identified in 12 (20%) JAK2(V617F)-positive or -negative patients. In a patient with 2 TET2 mutations, the analysis of 5 blood samples at different phases of her disease showed the sequential occurrence of JAK2(V617F) and TET2 mutations concomitantly to the disease evolution. Analysis of familial segregation confirmed that TET2 mutations were not inherited but somatically acquired. TET2 mutations were mainly observed (10 of 12) in patients with primary myelofibrosis or patients with polycythemia vera or essential thrombocythemia who secondarily evolved toward myelofibrosis or acute myeloid leukemia. PMID- 19564636 TI - Treatment of myelodysplastic syndrome patients with erythropoietin with or without granulocyte colony-stimulating factor: results of a prospective randomized phase 3 trial by the Eastern Cooperative Oncology Group (E1996). AB - This phase 3 prospective randomized trial evaluated the efficacy and long-term safety of erythropoietin (EPO) with or without granulocyte colony-stimulating factor plus supportive care (SC; n = 53) versus SC alone (n = 57) for the treatment of anemic patients with lower-risk myelodysplastic syndromes. The response rates in the EPO versus SC alone arms were 36% versus 9.6%, respectively, at the initial treatment step, 47% in the EPO arm, including subsequent steps. Responding patients had significantly lower serum EPO levels (45% vs 5% responses for levels < 200 mU/mL vs > or = 200 mU/mL) and improvement in multiple quality-of-life domains. With prolonged follow-up (median, 5.8 years), no differences were found in overall survival of patients in the EPO versus SC arms (median, 3.1 vs 2.6 years) or in the incidence of transformation to acute myeloid leukemia (7.5% and 10.5% patients, respectively). Increased survival was demonstrated for erythroid responders versus nonresponders (median, 5.5 vs 2.3 years). Flow cytometric analysis showed that the percentage of P glycoprotein(+) CD34(+) marrow blasts was positively correlated with longer overall survival. In comparison with SC alone, patients receiving EPO with or without granulocyte colony-stimulating factor plus SC had improved erythroid responses, similar survival, and incidence of acute myeloid leukemia transformation. PMID- 19564639 TI - Miltenberger blood group antigen type III (Mi.III) enhances the expression of band 3. AB - The special blood group antigen Mi.III exhibits a characteristic hybrid structure of glycophorin A (GPA) and glycophorin B, termed Gp.Mur. This phenotype has exceptionally high occurrence rates in several indigenous tribes in Taiwan ( approximately 21.2%-88.4%). Because glycophorin/Miltenberger begins interaction with anion exchanger-1 (AE1) in the endoplasmic reticulum, we hypothesized that the AE1-based macrocomplexes on erythrocyte membranes obtained from Mi.III(+) people could be differentiated from those obtained from non-Miltenberger people. Quantitative mass spectrometric comparison of the AE1-based complexes by iTRAQ (Applied Biosystems) revealed 25% to 67% higher expression of AE1 in Mi.III(+) erythrocytes. In accordance with the higher AE1 level, the Mi.III(+) erythrocytes exhibited superior HCO(3)(-) capacities, pH homeostasis, and osmotic resistance. Cotransfection experiments in HEK293 cells showed that Gp.Mur, like GPA, enhanced trafficking of AE1 to the plasma membrane. In summary, the increased surface expression of AE1 in Mi.III(+) erythrocytes could be attributed to the additive effect of GPA and Gp.Mur coexpression. PMID- 19564640 TI - Physician clinical information technology and health care disparities. AB - The authors develop a conceptual framework regarding how information technology (IT) can alter within-physician disparities, and they empirically test some of its implications in the context of coronary heart disease. Using a random experiment on 256 primary care physicians, the authors analyze the relationships between three IT functions (feedback and two types of clinical decision support) and five process-of-care measures. Endogeneity is addressed by eliminating unobserved patient characteristics with vignettes and by proxying for omitted physician characteristics. The results indicate that IT has no effects on physicians' diagnostic certainty and treatment of vignette patients overall. The authors find that treatment and certainty differ by patient age, gender, and race. Consistent with the framework, IT's effects on these disparities are complex. Feedback eliminated the gender disparities, but the relationships differed for other IT functions and process measures. Current policies to reduce disparities and increase IT adoption may be in discord. PMID- 19564641 TI - CETP does not affect triglyceride production or clearance in APOE*3-Leiden mice. AB - The cholesteryl ester transfer protein (CETP) facilitates the bidirectional transfer of cholesteryl esters and triglycerides (TG) between HDL and (V)LDL. By shifting cholesterol in plasma from HDL to (V)LDL in exchange for VLDL-TG, CETP aggravates atherosclerosis in hyperlipidemic APOE*3-Leiden (E3L) mice. The aim of this study was to investigate the role of CETP in TG metabolism and high-fat diet induced obesity by using E3L mice with and without the expression of the human CETP gene. On chow, plasma lipid levels were comparable between both male and female E3L and E3L.CETP mice. Further mechanistic studies were performed using male mice. CETP expression increased the level of TG in HDL. CETP did not affect the postprandial plasma TG response or the hepatic VLDL-TG and VLDL apolipoprotein B production rate. Moreover, CETP did not affect the plasma TG clearance rate or organ-specific TG uptake after infusion of VLDL-like emulsion particles. In line with the absence of an effect of CETP on tissue-specific TG uptake, CETP also did not affect weight gain in response to a high-fat diet. In conclusion, the CETP-induced increase of TG in the HDL fraction of E3L mice is not associated with changes in the production of TG or with tissue-specific clearance of TG from the plasma. PMID- 19564642 TI - Increased circulating levels of the antiangiogenic factor endostatin in early onset but not late-onset preeclampsia. AB - Changes in circulating angiogenic factors seem to play a key role in the pathogenesis of preeclampsia and it seems as if these changes are of greater importance in the pathogenesis of early-onset than of late-onset disease. Endostatin is a potent, broad spectrum antagonist of angiogenesis whose role in preeclampsia is poorly investigated. The aim of this study was to estimate whether circulating endostatin levels are altered in preeclampsia, and whether women with early-onset (before 32 weeks of gestation; n = 16) and late-onset (after 35 weeks of gestation; n = 19) preeclampsia differ in this regard. Women with early-onset, but not of late-onset preeclampsia had higher levels of endostatin than healthy pregnant women in corresponding lengths of gestation. The results of the study support the hypothesis that there is heterogeneity between early- and late-onset preeclampsia, with a stronger association between an altered angiogenic balance and early-onset than late-onset disease. PMID- 19564643 TI - Postpartum outcome of cervical intraepithelial neoplasia in pregnant women determined by route of delivery. AB - Cervical intraepithelial neoplasia (CIN) has its highest incidence during women's reproductive years. During 2 sequential 7-year periods, 1994 to 2000 and 2001 to 2007, 3695 and 3894 deliveries were performed, respectively, at Osaka University Hospital. CIN was detected in 21 cases (0.57%) during 1994-2000 and in 43 cases (1.1%) during 2001-2007. By comparison, cervical intraepithelial neoplasia complicated pregnancies increased significantly in the latter period (P = .015 by Fisher exact test, Odds ratio = 1.95; 95%CI: 1.16-3.30). We observed CIN regression in 34 (76%) of 45 cases of vaginal delivery and in 6 (50%) of 12 cases of cesarean delivery, indicating that the outcome of an initially diagnosed CIN and the delivery routes appeared not to be significantly related. However, a different result was obtained when only those patients whose CIN lesions persisted until the delivery were analyzed. Among the 35 such cases in the vaginal delivery group, 24 cases (69%) regressed after the delivery; in 8 such cases from the cesarean delivery group, only 2 cases (25%) regressed afterward. Our study clearly shows that pregnancy complicated with CIN is increasing rapidly in Japan. We also find that there is a significantly more frequent postpartum regression of biopsy-proven CIN lesions following a vaginal delivery compared to cesarean section (P = .042 by Fisher exact test, Odds ratio = 6.55; 95% CI: 1.13 37.8). PMID- 19564644 TI - The role of toll-like receptors (TLR-2 and -4) and triggering receptor expressed on myeloid cells 1 (TREM-1) in human term and preterm labor. AB - OBJECTIVE: To define the role of Toll-like receptors (TLR-2, TLR-4) and triggering receptor expressed on myeloid cells-1 (TREM-1) in pregnant human myometrium. STUDY DESIGN: Expression of TLR-2 and -4 mRNA and protein and TREM-1 mRNA was quantified in human myometrial samples. Subsequently, we investigated the effect of medroxyprogesterone acetate (MPA) as a functional inhibitor of the TLR agonist lipopolysaccharide (LPS). RESULTS: Messenger RNA expression of TLR-2 and -4 was significantly higher in myometrium at term compared with preterm (P = .009 and .016, respectively). Toll-like receptor-2 protein expression was significantly higher during labor (P = .002) compared with nonlaboring samples. Triggering receptor expressed on myeloid cells-1 mRNA expression was increased in both myometrium and cervix after labor at term (P < .05). Medroxyprogesterone acetate significantly suppressed LPS-induced production of interleukin 1b (IL 1b), IL-6, and IL-8 in vitro (P < .05). CONCLUSION: Toll-like receptors 2and 4 and TREM-1 are expressed in human myometrium and may play a role in the mechanism of labor at term, and their functional effects are inhibited by MPA. PMID- 19564645 TI - Neuroimaging patterns of central nervous system metastases in neuroblastoma: report of 2 recent cases and literature review. AB - The authors describe imaging patterns of intracranial metastases in 2 children with grade 4 neuroblastoma. Central nervous system metastases from neuroblastoma are extremely rare and may involve the cerebral parenchyma, leptomeninges, or dura. Cerebral parenchymal metastases can be cystic with mural nodules or solid with hemorrhagic elements. The first patient in our study had multiple cystic parenchymal metastases with calcific mural nodules, while the second patient developed solid hemorrhagic parenchymal metastatic lesions along with extensive leptomeningeal and dural deposits. Central nervous system involvement in both patients occurred within a time span ranging from 12 to 14 months from the time of initial diagnosis. PMID- 19564646 TI - Do vacuum-assisted deliveries cause intracranial vessel injuries? AB - Vacuum-assisted deliveries are fairly commonly used in obstetrical practice. Most newborns who have a vacuum-assisted delivery undergo extracranial birth traumas that have no residual consequences. Vacuum-assisted deliveries that complicate intracranial vascular infarction are rarely reported. We present 2 cases of intracranial vessel infarction after vacuum-assisted deliveries. One newborn, with scalp erosion, showed an unusual left middle cerebral artery infarct, and the other, with a severe subgaleal hematoma, had a venous thrombosis. Before the diagnosis, made using brain ultrasonography, neither had specific observable neurological symptoms. In conclusion, vacuum-assisted deliveries should be given special attention, especially when they are combined with a severe extracranial birth trauma. PMID- 19564647 TI - Celiac disease presenting as autism. AB - Gluten-restricted diets have become increasingly popular among parents seeking treatment for children diagnosed with autism. Some of the reported response to celiac diets in children with autism may be related to amelioration of nutritional deficiency resulting from undiagnosed gluten sensitivity and consequent malabsorption. A case is presented of a 5-year-old boy diagnosed with severe autism at a specialty clinic for autistic spectrum disorders. After initial investigation suggested underlying celiac disease and varied nutrient deficiencies, a gluten-free diet was instituted along with dietary and supplemental measures to secure nutritional sufficiency. The patient's gastrointestinal symptoms rapidly resolved, and signs and symptoms suggestive of autism progressively abated. This case is an example of a common malabsorption syndrome associated with central nervous system dysfunction and suggests that in some contexts, nutritional deficiency may be a determinant of developmental delay. It is recommended that all children with neurodevelopmental problems be assessed for nutritional deficiency and malabsorption syndromes. PMID- 19564648 TI - Metformin: safety in cardiac patients. AB - Metformin is a biguanide, insulin sensitiser that reduces blood sugar levels. There are concerns about the risk of lactic acidosis in patients receiving metformin who have procedures requiring iodinated contrast, and in those with renal impairment or heart failure. The data on which these concerns are based are reviewed, with the conclusion that metformin treatment is rarely to blame for lactic acidosis. A generic policy of stopping metformin 48 h before and 48 h after the procedure in all patients is counterintuitive, lacks any evidence base and does not conform to the principles of best practice. In patients with heart failure, although the underlying condition can predispose to lactic acidosis, existing evidence suggests that metformin use is associated with improved outcome rather than increased risk. PMID- 19564649 TI - Trends in HIV testing and recording of HIV status in the UK primary care setting: a retrospective cohort study 1995-2005. AB - OBJECTIVES: To provide nationally representative data on trends in HIV testing in primary care and to estimate the proportion of diagnosed HIV positive individuals known to general practitioners (GPs). METHODS: We undertook a retrospective cohort study between 1995 and 2005 of all general practices contributing data to the UK General Practice Research Database (GPRD), and data on persons accessing HIV care (Survey of Prevalent HIV Infections Diagnosed). We identified all practice-registered patients where an HIV test or HIV positive status is recorded in their general practice records. HIV testing in primary care and prevalence of recorded HIV positive status in primary care were estimated. RESULTS: Despite 11 fold increases in male testing and 19-fold increases in non-pregnant female testing between 1995 and 2005, HIV testing rates remained low in 2005 at 71.3 and 61.2 tests per 100,000 person years for males and females, respectively, peaking at 162.5 and 173.8 per 100,000 person years at 25-34 years of age. Inclusion of antenatal tests yielded a 129-fold increase in women over the 10-year period. In 2005, 50.7% of HIV positive individuals had their diagnosis recorded with a lower proportion in London (41.8%) than outside the capital (60.1%). CONCLUSION: HIV testing rates in primary care remain low. Normalisation of HIV testing and recording in primary care in antenatal testing has not been accompanied by a step change in wider HIV testing practice. Recording of HIV positive status by GPs remains low and GPs may be unaware of HIV-related morbidity or potential drug interactions. PMID- 19564650 TI - Detergent protease exposure and respiratory disease: case-referent analysis of a retrospective cohort. AB - OBJECTIVES: To examine the relationship between protease exposure and respiratory disease in a cohort of detergent enzyme manufacturers. METHODS: Case-referent analysis of a cohort of employees working in a European detergent factory between 1989 and 2002. Cases with new lower or upper respiratory disease were ascertained by examination of occupational health records and matched to referents on date of first employment. Personal exposures to airborne detergent protease were estimated, using a job exposure matrix, from >12,000 measurements taken in the factory during the period of study. RESULTS: We found clear, monotonic relationships between estimated protease exposure and both lower and upper respiratory disease. After control for age, sex and smoking, the odds ratio of lower respiratory disease was significantly elevated (1.98, 95% CI 1.04 to 3.79) in those employees working in jobs in the highest quartile of protease exposure (geometric mean 7.9 ng x m(-3)). For employees with upper respiratory disease, the risk was significantly elevated at a lower level of estimated protease exposure (geometric mean 2.3 ng x m(-3)). CONCLUSIONS: These findings provide strong evidence of an association between detergent enzyme exposure and the development of respiratory disease in an occupational setting. Using the routinely collected information on specific sensitisation and the close attention to workplace exposures that are characteristic of this industry, it should be possible to derive meaningful occupational exposure standards for most detergent enzymes. PMID- 19564651 TI - Work status, work hours and health in women with and without children. AB - OBJECTIVES: The authors studied self-reported health in women with and without children in relation to their work status (employed, student, job seeker or homemaker), work hours and having an employed partner. METHODS: The study group comprised of 6515 women born in 1960-1979 who were interviewed in one of the Swedish Surveys of Living Conditions in 1994-2003. Self-rated health, fatigue and symptoms of anxiety were analysed. RESULTS: Having children increased the odds of poor self-rated health and fatigue in employed women, female students and job seekers. The presence of a working partner marginally buffered the effects. In dual-earner couples, mothers reported anxiety symptoms less often than women without children. Few women were homemakers (5.8%). The odds of poor self-rated health and fatigue increased with increasing number of children in employed women, and in women working 40 h or more. Poor self-rated health was also associated with the number of children in students. Many mothers wished to reduce their working hours, suggesting time stress was a factor in their impaired health. The associations between having children and health symptoms were not exclusively attributed to having young children. CONCLUSIONS: Having children may contribute to fatigue and poor self-rated health particularly in women working 40 h or more per week. Student mothers and job seeking mothers were also at increased risk of poor self-rated health. The results should be noted by Swedish policy-makers. Also countries aiming for economic and gender equality should consider factors that may facilitate successful merging of work and family life. PMID- 19564652 TI - Constitutional (germline) MLH1 epimutation as an aetiological mechanism for hereditary non-polyposis colorectal cancer. AB - Hereditary non-polyposis colorectal cancer (HNPCC) is an autosomal dominant syndrome characterised by a predisposition to early onset colorectal, endometrial and other cancers. The tumours typically exhibit microsatellite instability due to defective mismatch repair. HNPCC is classically caused by heterozygous loss-of function mutations within the mismatch repair genes MLH1, MSH2, MSH6 and PMS2, but no pathogenic mutations are identified in a third of cases. In recent years, constitutional epimutations of the MLH1 gene, characterised by soma-wide allele specific promoter methylation and transcriptional silencing, have been identified in a handful of mutation negative HNPCC cases. In contrast to genetic mutations, MLH1 epimutations are reversible between generations and thus display non Mendelian inheritance. This review focuses on the aetiological role of constitutional MLH1 epimutations in the development of HNPCC related cancers. The molecular characteristics, clinical ramifications and potential mechanism underlying this defect are discussed. Recommendations for the selection of cases warranting screening for MLH1 epimutations are proffered. PMID- 19564654 TI - The physical activity transition. AB - Physical activity is important for the prevention of chronic disease morbidity and mortality, and the lack of adequate levels of physical activity represents a growing public health burden around the world. The purpose of this report is to introduce the concept of the "Physical Activity Transition" and to explore the potential effects that declining physical activity levels may play on health and life expectancy as countries undergo economic and demographic changes. Physical activity is related to mortality rates in humans, and the available evidence suggests that the adoption of a lifestyle characterized by lower levels of physical activity will attenuate the expected gains in life expectancy associated with the epidemiological transition. Advances in the measurement of physical activity at work, in the home, for transport, and in leisure time in a wide variety of populations will be integral to advancing the current understanding of how macro-level factors shape physical activity patterns and patterns of morbidity and mortality. PMID- 19564653 TI - Phenotypic variability in Rett syndrome associated with FOXG1 mutations in females. AB - BACKGROUND: The FOXG1 gene has been recently implicated in the congenital form of Rett syndrome (RTT). It encodes the fork-head box protein G1, a winged-helix transcriptional repressor with expression restricted to testis and brain, where it is critical for forebrain development. So far, only two point mutations in FOXG1 have been reported in females affected by the congenital form of RTT. Aim To assess the involvement of FOXG1 in the molecular aetiology of classical RTT and related disorders. METHODS: The entire multi-exon coding sequence of FOXG1 was screened for point mutations and large rearrangements in a cohort of 35 MECP2/CDKL5 mutation-negative female patients including 31 classical and four congenital forms of RTT. RESULTS: Two different de novo heterozygous FOXG1 truncating mutations were identified. The subject with the p.Trp308X mutation presented with a severe RTT-like neurodevelopmental disorder, whereas the p.Tyr400X allele was associated with a classical clinical RTT presentation. CONCLUSIONS: These new cases give additional support to the genetic heterogeneity in RTT and help to delineate the clinical spectrum of the FOXG1-related phenotypes. FOXG1 screening should be considered in the molecular diagnosis of RTT. PMID- 19564655 TI - Promotion of walking for transportation: a report from the Walk to School day registry. AB - BACKGROUND: In the US, promoting active transportation to school has received increased attention as a means of encouraging physical activity and preventing obesity among youth. However, little systematic evaluation of existing programs, such as Walk to School (WTS), has occurred. METHODS: WTS coordinators from across the US were surveyed (via web, mail, and telephone) about program activities, school and environmental characteristics, and perceived changes in children walking to school. As an exploratory aim, logistic regression analyses were used to examine program characteristics associated with perceived increases in children walking. RESULTS: From a database of 783 coordinators, 493 usable surveys were returned. Almost all respondents (98.2%) participated in a one-day WTS event. Other common activities included promotional activities (72.7%), safety trainings (49.6%), walkability audits (48.5%), and designated safe walking routes (46.5%). As part of their WTS efforts, 24.4% made policy changes and 38.4% made changes to the physical environment. Logistic regression analyses showed that policy changes, physical environment improvements, and number of activities were associated with the largest perceived increased in children walking to school. CONCLUSIONS: These findings help address the gap in knowledge about schools' participation in WTS programs, and suggest strategies to increase active transportation to school. PMID- 19564656 TI - Socioeconomic position and children's physical activity and sedentary behaviors: longitudinal findings from the CLAN study. AB - BACKGROUND: This study aimed to examine cross-sectional and longitudinal associations between socioeconomic position (SEP) and physical activity and sedentary behaviors amongst children and adolescents. METHODS: Maternal education was reported by parents of 184 5-6 year-old and 358 10-12 year-old children in 2001. In 2001 and 2004, physical activity was assessed by accelerometry. Older children self-reported and parents of younger children proxy-reported physical activity and television (TV) viewing behaviors. Linear regression was used to predict physical activity and sedentary behaviors, and changes in these behaviors, from maternal education. RESULTS: Among all children, accelerometer determined and self/parent-reported moderate and vigorous physical activity declined over three years. Girls of higher SEP demonstrated greater decreases in TV viewing behaviours than those of low SEP. In general, no prospective associations were evident between SEP and objectively-assessed physical activity. A small number of prospective associations were noted between SEP and self reported physical activity, but these were generally weak and inconsistent in direction. CONCLUSIONS: This study did not find strong evidence that maternal education was cross-sectionally or longitudinally predictive of children's physical activity or sedentary behaviors. Given the well-documented inverse relationship of SEP with physical activity levels in adult samples, findings suggest that such disparities may emerge after adolescence. PMID- 19564657 TI - Associations between after-school physical activity, television use, and parental strategies in a sample of New Zealand adolescents. AB - BACKGROUND: Youth display suboptimal levels of[1] physical activity and sedentary behaviors. Few studies have examined the existence of the "displacement hypothesis" or the effect of parental strategies on activity behaviors during the after-school period. METHODS: A total of 3471 students (12-18 years old) completed a self-report survey that assessed after-school physical activity and television (TV) use and perceived parental strategies (ie, encouragement to be active, TV-viewing rules). Participants were grouped into 4 activity groups: high TV/low active, high TV/active, low TV/low active, or low TV/active. Descriptive statistics and nominal logistic-regression analyses were conducted. RESULTS: Compared with students who watched less than 1 h of TV, participants who watched >or=4 h of TV were half as likely to be active after school (>or=4 h; adjusted odds ratio 0.51, 95% CI .40-.65). Compared with the low TV/active group, the other activity groups were at least 1.28 times more likely to have parents that provided only 1 parental strategy (encouragement for activity or TV rules) and up to 4.7 times more likely to have parents that provided neither strategy. DISCUSSION: Sedentary behaviors are associated with displacement of active pursuits. Parental strategies exert a strong influence on after-school behaviors of high school students. PMID- 19564658 TI - Individual and socio-ecological correlates of physical activity among Arab and Jewish college students in Israel. AB - BACKGROUND: Ample research has assessed correlates of physical activity (PA) among college students; however, socioenvironmental correlates of PA have not been assessed in Arab and Jewish Israeli college students. METHODS: Cross sectional study of 198 Arab and Jewish physical education college students. The dependent variable was meeting the CDC/ACSM guidelines for moderate/ vigorous PA. Independent variables consisted of individual variables (eg, ethnicity, gender, religious observance), and socioenvironmental variables (eg, street lighting, family support, exercise facilities). Bivariate and multivariate analyses were used. RESULTS: Thirty three percent of the students met the recommended guidelines for PA. Individual variables were more highly correlated with PA than socioenvironmental variables. In the final logistic regression model three individual covariates independently predicted PA: gender, race/ethnicity, and self-efficacy. Access to open space was the only environmental variable significantly correlated with PA. CONCLUSIONS: The results underscore the need for implementing an intervention program aimed at promoting the recommended levels of PA among Arab and Jewish Israeli physical education college students, while tailoring the intervention to individual risk markers for physical inactivity (eg, race/ethnicity, gender). PMID- 19564659 TI - Prevalence of physical activity among Lebanese adults: a cross-sectional study. AB - BACKGROUND: This survey aims to assess the prevalence of physical activity among adult Lebanese, and to report the relationship between sociodemographic variables and physical activity behavior highlighting the correlates discouraging people to carry out physical activity. METHODS: A cross-sectional study using an anonymous self-reported questionnaire was conducted on 346 adults from four Lebanese districts. Demographic characteristics, physical activity, smoking status, alcohol consumption, and medical history were obtained. RESULTS: Prevalence of physical activity among Lebanese adults was 55.5% (192/346). Age, BMI, marital status, medical history, occupation, educational level and smoking were significantly associated with physical activity (P < .05). Inactive obese participants were about three times more likely to report hypertension and diabetes than inactive normal weight participants (P = .013). BMI was significantly higher among inactive participants (P = .014). CONCLUSION: Physical activity was under level among Lebanese adults. Married, non office workers and smokers were the main correlates of physical inactivity in Lebanese adulthood. PMID- 19564660 TI - Testing compensation: does recreational basketball impact adult activity levels? AB - BACKGROUND: Current guidelines encourage adults to perform regular physical activity (PA) for optimal health, yet the majority of adults fail to meet the guidelines. One explanation for the difficulty in adding PA to meet recommended levels is an internal PA control center that may result in a compensatory lowering of normal activity levels after "added activity" sessions during the day. The purpose of this study was to test the compensation hypothesis by assessing PA on days of "added" PA among adults. METHODS: Twenty middle-aged adults recorded daily step counts, in addition to step counts and minutes of basketball play. To test for compensation, step counts on non basketball days were compared with steps counts on basketball days (excluding basketball steps). RESULTS: No significant differences (F = 0.711) were found between groups. In summary, no compensatory decrease in PA was identified on basketball participation days in this population. When steps in basketball were added, differences (P = .01) in daily step counts existed between basketball days (mean = 15,568) and non basketball days (mean = 8,408). CONCLUSION: These results suggest that "added" PA (basketball) does not result in compensatory reductions in typical daily PA on days of "added" activity for the population studied. PMID- 19564661 TI - Walking speed at self-selected exercise pace is lower but energy cost higher in older versus younger women. AB - BACKGROUND: Walking is usually undertaken at a speed that coincides with the lowest metabolic cost. Aging however, alters the speed-cost relationship, as preferred walking speeds decrease and energy costs increase. It is unclear to what extent this relationship is affected when older women undertake walking as an exercise modality. The aim of this study was to compare the energetic cost of walking at a self-selected exercise pace for a 30 minute period in older and younger women. METHODS: The energetic cost of walking was assessed using the energy equivalent of oxygen consumption measured in 18 young (age 25 to 49 years) and 20 older (age 50 to 79 years) women who were asked to walk at their 'normal' exercise pace on a motorized treadmill for 30 minutes duration. RESULTS: The mass specific net cost of walking (Cw) was 15% higher and self-selected walking speed was 23% lower in the older women than in the younger group. When speed was held constant, the Cw was 0.30 (J.kg-1.m-1) higher in the older women. CONCLUSIONS: Preferred exercise pace incurs a higher metabolic cost in older women and needs be taken into consideration when recommending walking as an exercise modality. PMID- 19564662 TI - Cardiovascular and metabolic responses to water aerobics exercise in middle-age and older adults. AB - BACKGROUND: The purpose of this study was (a) to assess the cardiovascular and metabolic responses to water aerobic exercise and (b) to determine if water aerobics exercise meets the American College of Sports Medicine (ACSM) guidelines for improving and maintaining cardiorespiratory fitness. METHODS: Fourteen men and women (mean +/- SD age, height, weight, body fat percentage, and maximal oxygen uptake (VO2max): = 57.4 +/- 7.6 years, 171.3 +/- 7.8 cm, 89.9 +/- 13.9 kg, 32.5 +/- 5.8%, and 31.0 +/- 8.3 mL/kg/min, respectively) completed a maximal treadmill exercise test and 50-min water aerobics session. Cardiovascular and metabolic data were collected via a portable calorimetric measurement system. RESULTS: Mean exercise intensity was 43.4% and 42.2% of heart rate reserve (HRR) and maximal oxygen uptake reserve (VO2R), respectively. Training intensity in metabolic equivalents (METS) was 4.26 +/- 0.96. Total net energy expenditure for the exercise session was 249.1 +/- 94.5 kcal/session. CONCLUSIONS: Results indicate that water aerobics is a feasible alternative to land-based exercise for middle-aged and older adults that fulfills the ACSM guidelines for improving and maintaining cardiorespiratory fitness. PMID- 19564664 TI - Linking the American Time Use Survey (ATUS) and the Compendium of Physical Activities: methods and rationale. AB - BACKGROUND: The 2003 Bureau of Labor Statistics American Time Use Survey (ATUS) contains 438 distinct primary activity variables that can be analyzed with regard to how time is spent by Americans. The Compendium of Physical Activities is used to code physical activities derived from various surveys, logs, diaries, etc to facilitate comparison of coded intensity levels across studies. METHODS: This article describes the methods, challenges, and rationale for linking Compendium estimates of physical activity intensity (METs, metabolic equivalents) with all activities reported in the 2003 ATUS. RESULTS: The assigned ATUS intensity levels are not intended to compute the energy costs of physical activity in individuals. Instead, they are intended to be used to identify time spent in activities broadly classified by type and intensity. This function will complement public health surveillance systems and aid in policy and health-promotion activities. For example, at least one of the future projects of this process is the descriptive epidemiology of time spent in common physical activity intensity categories. CONCLUSIONS: The process of metabolic coding of the ATUS by linking it with the Compendium of Physical Activities can make important contributions to our understanding of American's time spent in health-related physical activity. PMID- 19564663 TI - A population-based study of the determinants of physical activity in ovarian cancer survivors. AB - BACKGROUND: Regular physical activity is associated with quality of life and other health-related outcomes in ovarian cancer survivors, but participation rates are low. This study investigated the determinants of physical activity in ovarian cancer survivors. METHODS: A population-based, cross-sectional, mailed survey of ovarian cancer survivors in Alberta, Canada, was performed. Measures included self-reported physical activity, medical and demographic factors, and social cognitive variables from the Theory of Planned Behavior. RESULTS: A total of 359 women participated (51.4% response rate), of whom 112 (31.1%) were meeting physical activity guidelines. Variables associated with meeting guidelines were younger age, higher education and income, being employed, lower body mass index, absence of arthritis, longer time since diagnosis, earlier disease stage, and being disease-free. Analysis of the Theory of Planned Behavior variables indicated that 36% of the variance in physical activity guidelines was explained, with intention being the sole independent correlate (?=.56; P < .001). CONCLUSION: Various demographic and medical factors can help identify ovarian cancer survivors at risk for physical inactivity. Interventions should attempt to increase physical activity intentions in this population by focusing on instrumental and affective attitudes as well as perceptions of control. PMID- 19564665 TI - Procedures used to standardize data collected by RT3 triaxial accelerometers in a large-scale weight-loss trial. AB - BACKGROUND: Accelerometers measure intensity, frequency, and duration of physical activity. However, the scarcity of reports on data reduction makes comparing accelerometer results across studies difficult. METHODS: Participants were asked to wear a triaxial accelerometer (RT3) for ?10 hours for at least 4 days, including one weekend day. We summarize our data-cleaning procedures and assess the impact of defining a usable day of measurements as at least 6, 8, or 10 hours of wear time, and of standardizing data to a 12-hour day. RESULTS: Eighty-two percent of participants met wear time requirements; 93% met requirements when we defined a day as 8-or-more hours of wear time. Normalization of data to a 12-hour day had little impact on estimates of daily moderate-to-vigorous physical activity (MVPA; 16.9 vs. 17.1 minutes); restricting MVPA to activities occurring in bouts of 10 minutes or longer had greater impact (16.9 vs. 6.3 minutes per day). CONCLUSION: Our account of accelerometry quality-control and data-cleaning procedures documents the small impact of variations in daily wear time requirements on MVPA estimates, and the larger impact of evaluating total MVPA vs. MVPA occurring in extended bouts. This paper should allow other researchers to duplicate or revise our methods as needed. PMID- 19564666 TI - A methodological model for collecting high-quality data on physical activity in developing settings-the experience of the 1993 Pelotas (Brazil) Birth Cohort study. AB - BACKGROUND: Prospective studies on physical activity (PA), diet, and body composition in adolescents are lacking, particularly outside high-income countries. GOALS: To describe the methods used to assess these variables in the 1993 Pelotas (Brazil) Birth Cohort and to discuss the fieldwork challenges faced and alternatives to overcome them. METHODS: In 2006-07 a subsample of the 1993 Pelotas cohort was revisited. PA was estimated using questionnaires, a combined heart-rate and motion sensor (Acti-Heart), and the ActiGraph GT1M accelerometer. Diet was investigated by questionnaire. Total body water was determined by stable isotopes. Thirty individuals had their total energy expenditure assessed by doubly labeled water. All data were collected at participants' home. RESULTS: The logistics of the fieldwork and the difficulties in undertaking the study and alternatives to overcome them are presented. Preliminary analyses show that 511 individuals were traced (response rate = 90.0%). Compliance of both adolescents and their families for the motion sensors and body-composition measurements was excellent. CONCLUSIONS: The authors conclude that it is feasible to carry out high-quality studies on PA in developing countries. They hope the article will be useful to other researchers interested in carrying out similar studies. PMID- 19564667 TI - Testing the reliability of neighborhood-specific measures of physical activity among Canadian adults. AB - BACKGROUND: Capturing neighborhood-specific physical activity is necessary for advancing understanding about the relations between neighborhood walkability and physical activity. This study examined the test-retest reliability of previously developed items (from the Neighborhood Physical Activity Questionnaire) for capturing setting-specific physical activity among Canadian adults. METHODS: Randomly sampled adults (n = 117) participated in two telephone-interviews 2 to 5 days apart. Respondents were asked a series of items capturing frequency and duration of transportation-related walking, recreational walking, moderate and vigorous-intensity physical activity undertaken inside and outside the neighborhood in a usual week. The test-test reliability of reported physical activity levels were then examined using Intraclass and Spearman's rank correlations, kappa coefficients, and overall agreement. RESULTS: Participation, frequency, and the duration of transportation-related and recreational walking and vigorous-intensity physical activity inside and outside the neighborhood showed moderate to excellent test-retest reliability. Moderate reliability was found for moderate-intensity physical activity undertaken inside (k = 0.48; ICC frequency = 0.38; ICC duration = 0.39) and outside (k = 0.51; ICC frequency = 0.79; ICC duration = 0.31) the neighborhood. CONCLUSIONS: Neighborhood-specific physical activity items administered by telephone-interview are reliable and are therefore appropriate for use in future studies that examining neighborhood walkability and physical activity. PMID- 19564668 TI - Development of a pedestrian walkability database of Northern Kentucky using Geographic Information Systems (GIS). AB - BACKGROUND: Rising obesity rates in the United States has spurred efforts by health advocates to encourage more active lifestyles including walking. Ensuring the availability, quality and safety of pedestrian walkways has become an important issue for government at all levels. METHODS: Pedestrian paths in Campbell County Kentucky were evaluated using a ranking criteria developed by the Walking and Bicycling Suitability Assessment (WABSA) project at the University of North Carolina School of Public Health. A pedestrian path Geographic Information System (GIS) data-layer was created and mobile GIS units were used to assess the sidewalk segments using the ranking. Data from sidewalk surveys were compared with Census 2000 blockgroup information on age of housing, population density and household transportation characteristics to examine the correlation between these factors and sidewalk presence and quality. The analysis explored to use of census data to predict walkability factors and looked for trends in quality and availability of pedestrian paths over time. RESULTS: Results showed higher overall scores for older urban areas adjacent to the Ohio River and Cincinnati. Housing built in the 1970s and 1980s showed the lowest scores while more recent housing showed improvement over earlier decades. Age of housing was determined to be a useful predictor while economic and population density attributes showed no correlation with walkability factors. CONCLUSION: Census housing age data are the most useful predictor of walkability demonstrating clear trends over time. The study shows improvements in walkways availability over the past few decades. However infrastructure improvements are needed to provide more extensive pedestrian walkways and linkages between existing walkways in Campbell County. PMID- 19564669 TI - Priority of activity-friendly community issues among key decision makers in Hawaii. AB - BACKGROUND: The U.S. Community Guide to Preventive Services strongly recommends changes in urban design, land use, and accessibility to increase physical activity. To achieve these goals, policy change is often needed. This study assessed attitudes of decision makers in Hawaii to determine if physical activity related issues are among their priorities. METHODS: State and county officials (n = 179) were mailed surveys. Respondents listed the three most important problems (open-ended) in Hawaii and rated the importance of 23 specified problems, of which six directly related to physical activity. RESULTS: The survey was completed by 126 (70.4%) respondents. The most frequently mentioned categories for the open-ended questions were affordable housing, environment/sustainability, sprawl/traffic/population growth, and healthcare. Among the closed-ended physical activity related items, increasing traffic was ranked highest (43.9%) and fourth overall. Less than 12% of decision makers rated other physical activity issues as important. CONCLUSIONS: Future work is needed to increase the visibility and importance of physical activity related issues among policymakers. PMID- 19564670 TI - Intermittent NPWT and lower negative pressures--exploring the disparity between science and current practice: a review. AB - Negative pressure wound therapy (NPWT) is used to treat a wide and growing range of problematic acute and chronic wounds. Continuous therapy delivered at -125 mm Hg has been routinely recommended, despite consistent research findings suggesting potential advantages to the use of lower pressures and intermittent therapy. To enhance understanding and document the disparity between the body of NPWT science and current practice with respect to negative pressure levels and modes of therapy, a search of the English-language literature from June 1989 through May 2009 was conducted. Thirty-six publications found to contain directly relevant information (in vitro, in vivo, and clinical data) were examined. While lower negative pressures and intermittent therapy were associated in earlier studies with improved microvascular blood flow in porcine wound models and with reduced pain in patients, early system shortcomings discourage adoption of intermittent therapy. Subsequent preclinical studies confirmed the beneficial effects of intermittent therapy compared to continuous therapy on blood flow and granulation tissue formation and lower pressures (-75 mm Hg or -100 mm Hg) compared to higher pressure (-125 mm Hg) on soft tissue blood flow. Considering the available preclinical evidence, reported patient pain, and common use of high pressure continuous NPWT in clinical practice, high-quality randomized controlled clinical trials must be conducted to help clinicians optimize care. PMID- 19564671 TI - Using low pressure, NPWT for wound preparation & the management of split thickness skin grafts in 3 patients with complex wound. AB - The use of negative pressure wound therapy (NPWT) is well established in the management of hard-to-heal wounds. One institution, familiar with NPWT's capabilities as well as its shortcomings (eg, pain at dressing changes and pain with the maximum recommended setting of 125 mm Hg), sought a viable alternative. A low pressure, negative pressure wound therapy (LP-NPWT) system, using subatmospheric pressure levels of 75 mm Hg and a low-adherence dressing, was evaluated to prepare the wound bed for split-thickness skin graft (STSG) on three patients. One patient was a healthy 23-year-old man with extensive trauma-related soft tissue wounds. The two women--54 and 47 years old--had multiple comorbidities. One had a lower extremity fasciotomy wound and the other had a dehisced surgical wound with a history of irradiation. Wound area was reduced >60% in all three wounds in 3 to 6 weeks as new granulation tissue developed. The average pain reported was moderate (4 to 5 on a Visual Analogue Scale), ranging from 2 to 10 during dressing changes to 1 to 7 between dressing changes; pain levels reported tended to decrease as therapy progressed. Little or no trauma on dressing removal and no signs of infection were noted. In all cases, STSGs, followed by 4 days of LP-NPWT were applied and all wounds healed. The results from these three cases suggest that the LP-NPWT system is a useful healing adjunct for complex wound bed preparation and graft management. Clinical studies to quantify the effects of LP-NPWT technology and compare its safety and efficacy to other negative pressure systems are needed. PMID- 19564672 TI - Stomal mucocutaneous dehiscence as a complication of topical negative pressure used to treat an open abdomen: a case series. AB - Although mucocutaneous dehiscence in cases of laparostomies is rare, this complication occurred in three patients in The Netherlands (all men, 53 to 65 years old) who received topical negative pressure (TNP) therapy in the management of open abdomens. Two patients experienced anastomotic failure following colonic resection and one had iatrogenic suturing of the small bowel to the fascia. TNP therapy was initiated and stomal dehiscence occurred after 2 to 3 days, contaminating the area. All patients developed sepsis and one died. Two required numerous additional surgeries and antibiotic treatments for 6 and 8 months, respectively. The complication was believed to have occurred as a result of tension on the proximal bowel of the stoma (the bowel inside the abdomen) not due to tension on the sutures or the small part of the bowel outside the abdomen. A modified TNP procedure, consisting of careful placement of the primary dressing around the stoma using separate primary dressing flaps, has since been used without complications in one patient. With implementation of TNP increasing and because of concerns regarding its use, additional research and innovative, simple solutions to manage complex wounds such as the one presented here are needed. PMID- 19564673 TI - Implementing wound care guidelines: observations and recommendations from the bedside. AB - The successful implementation of wound care guidelines requires an appreciation for the frustrations experienced by nurses trying to incorporate these tools into clinical practice. These frustrations or barriers to best wound care practice implementation are examined from the perspective of: 1) the practice environment, which must be understood; 2) the potential adopters, predominantly nurses seeking the best fit between evidence and their clinical practice setting; and 3) the evidence-based innovation created to change wound care practice at the point of care. Barriers identified include lack of available resources, time constraints, prescriptive guidelines that incorrectly assume details of the practice environment, and wound care product confusion. Recommendations to facilitate implementation from the bedside are discussed and include expanding guidelines to incorporate detailed educational content and dissemination strategies that serve to increase relevancy to everyday practice. Additional suggestions include decreasing wound care product confusion by developing standardized, function based product nomenclature and improving the quality of wound care research to increase nurses' confidence in the evidence and resultant recommendations. Resources currently used to develop guidelines also should be utilized to create accompanying educational material to support the transfer and uptake of knowledge. PMID- 19564674 TI - Cortical mechanisms of normal and abnormal processing in the visual system, Part 1. Spatial vision, amblyopia, hyperacuity, modal assumptions: a review. AB - Vernier thresholds rise much more rapidly in the periphery than do grating thresholds. A similar dissociation between acuity and hyperacuity has been shown to be present in strabismic but not in anisometropic amblyopia, thus leading to the suggestion that the strabismic fovea is similar to the normal periphery. Here it is shown that a quantitative model for spatial pattern discrimination, which accounts for foveal hyperacuity data, can be extended to encompass both the periphery and amblyopia if appropriate alterations are made. For the periphery it is necessary to increase the size of model receptive fields and to introduce both spatial undersampling and position irregularity (i.e. irregularity in the location of cortical filters). The strabismic fovea also requires spatial undersampling and position irregularity but no change in receptive field size. Defects in the good eye of strabismus can be explained by spatial irregularity. Finally, the anisometropic fovea requires a reduction in mechanism sensitivity but neither spatial untersampling nor position irregularity. PMID- 19564675 TI - Cortical mechanisms of normal and abnormal processes in the visual system, Part 2. Cortical regions involved in perceiving object shape: a review. PMID- 19564676 TI - Cortical mechanisms of normal and abnormal processes in the visual system, Part 3. Neurophysiological evidence for contrast dependent long-range facilitation and suppression in the human visual cortex: a review. PMID- 19564677 TI - An internet-based wearable watch-over system for elderly and disabled utilizing EMG and accelerometer. AB - An effective way for preventing injuries and diseases among the elderly is to monitor their daily lives. In this regard, we propose the use of a "Hyper Hospital Network", which is an information support system for elderly people and patients. In the current study, we developed a wearable system for monitoring electromyography (EMG) and acceleration using the Hyper Hospital Network plan. The current system is an upgraded version of our previous system for gait analysis (Yoshida et al. [13], Telemedicine and e-Health 13 703-714), and lets us monitor decreases in exercise and the presence of a hemiplegic gait more accurately. To clarify the capabilities and reliability of the system, we performed three experimental evaluations: one to verify the performance of the wearable system, a second to detect a hemiplegic gait, and a third to monitor EMG and accelerations simultaneously. Our system successfully detected a lack of exercise by monitoring the iEMG in healthy volunteers. Moreover, by using EMG and acceleration signals simultaneously, the reliability of the Hampering Index (HI) for detecting hemiplegia walking was improved significantly. The present study provides useful knowledge for the development of a wearable computer designed to monitor the physical conditions of older persons and patients. PMID- 19564678 TI - Limitations and pitfalls of 3-D fluoroscopic navigation in orthopaedic trauma surgery. PMID- 19564679 TI - An internet based learning tool in orthopaedic surgery: preliminary experiences and results. AB - Today universities can offer a variety of educational resources to their students through the internet. These may include lecture notes, PowerPoint presentations, or even an entire recording of a lecture in video format. At the Hannover Medical School (Hannover, Germany), the Trauma Surgery Department in collaboration with the Institute of Medical Informatics has developed an alternative method of "E learning". We created a web-based multimedia resource center for Trauma Surgery using the Content-Management-System (CMS) Schoolbook application, which was initially developed by the Institute of Medical Informatics. The so called "Trauma Surgery Schoolbook" was first adopted in October of 2005 and has since been used and evaluated by medical students at our institution. The evaluation results for the academic year 2005/06 are reported in this paper. The majority of students enrolled in the Trauma Surgery rotation utilized the Schoolbook, which they regarded as a helpful and effective study tool. Our students embraced the possibility of being able to prepare for lectures and use the Schoolbook for independent home studying purposes. Over time, there was a steady increase in the utilization of the Schoolbook by the students from 67% in the first trimester to 93% in the third trimester. The majority of the surveyed students (79.6%) found the Schoolbook to be constructive and helpful. 8.1% did not have any opinion, and only 12.3% found it to not be helpful. The instructors also found this web-based training program to be both constructive and practical, and were able to utilize its multimedia components to complement their lectures. Overall, our experience with this computer-aided learning program demonstrated that web-based technologies can improve the quality of medical education, benefiting both the students and the instructors. PMID- 19564680 TI - Direct cellular vs. indirect pager communication during orthopaedic surgical procedures: a prospective study. AB - BACKGROUND: Cellular phone use within the hospital setting has increased as physicians, nurses, and ancillary staff incorporate wireless technologies in improving efficiencies, cost, and maintaining patient safety and high quality healthcare [11]. Through the use of wireless, cellular communication, an overall improvement in communication accuracy and efficiency between intraoperative orthopaedic surgeons and floor nurses may be achieved. METHODS: Both communication types occurred while the surgeon was scrubbed in the operating room (OR). Indirect communication occurred when the pager call was answered by the OR circulating nurse with communication between the surgeon, circulating nurse, and floor nurse. Direct communication consisted of cell phone and Jabra Bluetooth BT200 wireless ear piece used by the surgeon. The surgeon answered the floor nurse's cellular call by phone ring-activated automatic answering. The study was conducted during scheduled orthopaedic procedures. An independent observer measured time variables with a stop-watch while orthopaedic nurses randomly called via pager or cell phone. The nurses asked for patient caregiver confirmation and answers to 30 different patient-care questions. RESULTS: Sixty trials were performed with 30 cell and 30 page communications. Direct cellular communication showed a better response rate than indirect page (Cell 100%, Page 73%). Indirect page communication allowed a 27% and 33% error rate with patient problem and surgeon solution communications, respectively. There were no reported communication errors while using direct wireless, cellular communication. When compared to page communications, cellular communications showed statistically significant improvements in mean time intervals in response time (Cell = 11s, Page = 211s), correct patient identification (Cell = 5s, Page = 172s), patient problem and solution time (Cell = 13s, Page = 189s), and total communication time (Cell = 32s, Page = 250s) (s = seconds, all P < 0.001). Floor nurse satisfaction ratings (dependent on communication times and/or difficulties) were improved with direct cellular communication (Cell = 29 excellent, Page = 11 excellent). Intraoperative case interruptions (defined as delaying surgical progress) were more frequent with indirect page communication (10 page v. 0 cell). CONCLUSION AND SIGNIFICANCE: Our study demonstrates that direct wireless communication may be used to improve intraoperative communication and enhance patient safety. Direct wireless, cellular intraoperative communication improves communication times, communication accuracy, communication satisfaction, and minimizes intraoperative case interruption. As a result of this study, we hope to maintain our transition to direct wireless, cellular intraoperative orthopaedic communication to reduce medical errors, improve patient care, and enhance both orthopaedic surgeon and nursing efficiencies. PMID- 19564681 TI - The Max-Inf2/Lorentz Center workshop on New algorithms in macromolecular crystallography and electron microscopy. AB - The resolution gap between macromolecular crystallography and electron microscopy continues to decrease. Recent advances in specimen preparation, instrumentation and computational power have allowed accurate structure determination of larger macromolecular complexes by crystallography and/or by electron microscopy on cryovitrified samples. New possibilities in structural biology have opened up and new challenges are faced to further reduce the resolution gap. A workshop at the Lorentz Center, Leiden, The Netherlands, which took place in May 2008, was organized to push further the limits of both complementary techniques through improved computational methods. PMID- 19564682 TI - Unit-cell determination from randomly oriented electron-diffraction patterns. AB - Unit-cell determination is the first step towards the structure solution of an unknown crystal form. Standard procedures for unit-cell determination cannot cope with data collections that consist of single diffraction patterns of multiple crystals, each with an unknown orientation. However, for beam-sensitive nanocrystals these are often the only data that can be obtained. An algorithm for unit-cell determination that uses randomly oriented electron-diffraction patterns with unknown angular relationships is presented here. The algorithm determined the unit cells of mineral, pharmaceutical and protein nanocrystals in orthorhombic high- and low-symmetry space groups, allowing (well oriented) patterns to be indexed. PMID- 19564683 TI - Application of normal-mode refinement to X-ray crystal structures at the lower resolution limit. AB - The structural refinement of large complexes at the lower resolution limit is often difficult and inefficient owing to the limited number of reflections and the frequently high-level structural flexibility. A new normal-mode-based X-ray crystallographic refinement method has recently been developed that enables anisotropic B-factor refinement using a drastically smaller number of thermal parameters than even isotropic refinement. Here, the method has been systematically tested on a total of eight systems in the resolution range 3.0-3.9 A. This series of tests established the most applicable scenarios for the method, the detailed procedures for its application and the degree of structural improvement. The results demonstrated substantial model improvement at the lower resolution limit, especially in cases in which other methods such as the translation-libration-screw (TLS) model were not applicable owing to the poorly converged isotropic B-factor distribution. It is expected that this normal-mode based method will be a useful tool for structural refinement, in particular at the lower resolution limit, in the field of X-ray crystallography. PMID- 19564684 TI - Cluster analysis for phasing with molecular replacement: a feasibility study. AB - Molecular replacement can fail to find a solution, namely a unique orientation and position of a search model, even when many search models are tested under various conditions. Simultaneous use of the results of these searches may help in the solution of such difficult structures. A closeness between the peaks of several calculated rotation functions may identify the model orientation. The largest and most compact cluster of such peaks usually corresponds to models which are oriented similarly to the molecule under study. A search for the optimal translation may be more problematic and both individual translation functions and straightforward cluster analysis in the space of geometric parameters such as rotation angles and translation vectors may give no result. An improvement may be obtained by performing cluster analysis of the peaks of several translation functions in phase-set space. In this case, the Fourier maps computed using the observed structure-factor magnitudes and the phases calculated from differently positioned models are compared. Again, as a rule, the largest and the most compact cluster corresponds to the correct solution. The result of the updated procedure is no longer a single search model but an averaged Fourier map. PMID- 19564686 TI - Parallel, distributed and GPU computing technologies in single-particle electron microscopy. AB - Most known methods for the determination of the structure of macromolecular complexes are limited or at least restricted at some point by their computational demands. Recent developments in information technology such as multicore, parallel and GPU processing can be used to overcome these limitations. In particular, graphics processing units (GPUs), which were originally developed for rendering real-time effects in computer games, are now ubiquitous and provide unprecedented computational power for scientific applications. Each parallel processing paradigm alone can improve overall performance; the increased computational performance obtained by combining all paradigms, unleashing the full power of today's technology, makes certain applications feasible that were previously virtually impossible. In this article, state-of-the-art paradigms are introduced, the tools and infrastructure needed to apply these paradigms are presented and a state-of-the-art infrastructure and solution strategy for moving scientific applications to the next generation of computer hardware is outlined. PMID- 19564685 TI - UROX 2.0: an interactive tool for fitting atomic models into electron-microscopy reconstructions. AB - Electron microscopy of a macromolecular structure can lead to three-dimensional reconstructions with resolutions that are typically in the 30-10 A range and sometimes even beyond 10 A. Fitting atomic models of the individual components of the macromolecular structure (e.g. those obtained by X-ray crystallography or nuclear magnetic resonance) into an electron-microscopy map allows the interpretation of the latter at near-atomic resolution, providing insight into the interactions between the components. Graphical software is presented that was designed for the interactive fitting and refinement of atomic models into electron-microscopy reconstructions. Several characteristics enable it to be applied over a wide range of cases and resolutions. Firstly, calculations are performed in reciprocal space, which results in fast algorithms. This allows the entire reconstruction (or at least a sizeable portion of it) to be used by taking into account the symmetry of the reconstruction both in the calculations and in the graphical display. Secondly, atomic models can be placed graphically in the map while the correlation between the model-based electron density and the electron-microscopy reconstruction is computed and displayed in real time. The positions and orientations of the models are refined by a least-squares minimization. Thirdly, normal-mode calculations can be used to simulate conformational changes between the atomic model of an individual component and its corresponding density within a macromolecular complex determined by electron microscopy. These features are illustrated using three practical cases with different symmetries and resolutions. The software, together with examples and user instructions, is available free of charge at http://mem.ibs.fr/UROX/. PMID- 19564687 TI - Introducing robustness to maximum-likelihood refinement of electron-microscopy data. AB - An expectation-maximization algorithm for maximum-likelihood refinement of electron-microscopy images is presented that is based on fitting mixtures of multivariate t-distributions. The novel algorithm has intrinsic characteristics for providing robustness against atypical observations in the data, which is illustrated using an experimental test set with artificially generated outliers. Tests on experimental data revealed only minor differences in two-dimensional classifications, while three-dimensional classification with the new algorithm gave stronger elongation factor G density in the corresponding class of a structurally heterogeneous ribosome data set than the conventional algorithm for Gaussian mixtures. PMID- 19564688 TI - Confidence intervals for fitting of atomic models into low-resolution densities. AB - The fitting of high-resolution structures into low-resolution densities obtained from techniques such as electron microscopy or small-angle X-ray scattering can yield powerful new insights. While several algorithms for achieving optimal fits have recently been developed, relatively little effort has been devoted to developing objective measures for judging the quality of the resulting fits, in particular with regard to the danger of overfitting. Here, a general method is presented for obtaining confidence intervals for atomic coordinates resulting from fitting of atomic resolution domain structures into low-resolution densities using well established statistical tools. It is demonstrated that the resulting confidence intervals are sufficiently accurate to allow meaningful statistical tests and to provide tools for detecting potential overfitting. PMID- 19564689 TI - Interpretation of very low resolution X-ray electron-density maps using core objects. AB - A novel approach to obtaining structural information from macromolecular X-ray data extending to resolutions as low as 20 A is presented. Following a simple map segmentation procedure, the approximate shapes of the domains forming the structure are identified. A pattern-recognition comparative analysis of these shapes and those derived from the structures of domains from the PDB results in candidate structural models that can be used for a fit into the density map. It is shown that the placed candidate models can be employed for subsequent phase extension to higher resolution. PMID- 19564691 TI - Structures of dihydrofolate reductase-thymidylate synthase of Trypanosoma cruzi in the folate-free state and in complex with two antifolate drugs, trimetrexate and methotrexate. AB - The flagellate protozoan parasite Trypanosoma cruzi is the pathogenic agent of Chagas disease (also called American trypanosomiasis), which causes approximately 50,000 deaths annually. The disease is endemic in South and Central America. The parasite is usually transmitted by a blood-feeding insect vector, but can also be transmitted via blood transfusion. In the chronic form, Chagas disease causes severe damage to the heart and other organs. There is no satisfactory treatment for chronic Chagas disease and no vaccine is available. There is an urgent need for the development of chemotherapeutic agents for the treatment of T. cruzi infection and therefore for the identification of potential drug targets. The dihydrofolate reductase activity of T. cruzi, which is expressed as part of a bifunctional enzyme, dihydrofolate reductase-thymidylate synthase (DHFR-TS), is a potential target for drug development. In order to gain a detailed understanding of the structure-function relationship of T. cruzi DHFR, the three-dimensional structure of this protein in complex with various ligands is being studied. Here, the crystal structures of T. cruzi DHFR-TS with three different compositions of the DHFR domain are reported: the folate-free state, the complex with the lipophilic antifolate trimetrexate (TMQ) and the complex with the classical antifolate methotrexate (MTX). These structures reveal that the enzyme is a homodimer with substantial interactions between the two TS domains of neighboring subunits. In contrast to the enzymes from Cryptosporidium hominis and Plasmodium falciparum, the DHFR and TS active sites of T. cruzi lie on the same side of the monomer. As in other parasitic DHFR-TS proteins, the N-terminal extension of the T. cruzi enzyme is involved in extensive interactions between the two domains. The DHFR active site of the T. cruzi enzyme shows subtle differences compared with its human counterpart. These differences may be exploited for the development of antifolate-based therapeutic agents for the treatment of T. cruzi infection. PMID- 19564690 TI - What can we learn by computing 13Calpha chemical shifts for X-ray protein models? AB - The room-temperature X-ray structures of ubiquitin (PDB code 1ubq) and of the RNA binding domain of nonstructural protein 1 of influenza A virus (PDB code 1ail) solved at 1.8 and 1.9 A resolution, respectively, were used to investigate whether a set of conformations rather than a single X-ray structure provides better agreement with both the X-ray data and the observed 13Calpha chemical shifts in solution. For this purpose, a set of new conformations for each of these proteins was generated by fitting them to the experimental X-ray data deposited in the PDB. For each of the generated structures, which show R and Rfree factors similar to those of the deposited X-ray structure, the 13Calpha chemical shifts of all residues in the sequence were computed at the DFT level of theory. The sets of conformations were then evaluated by their ability to reproduce the observed 13Calpha chemical shifts by using the conformational average root-mean-square-deviation (ca-r.m.s.d.). For ubiquitin, the computed set of conformations is a better representation of the observed 13Calpha chemical shifts in terms of the ca-r.m.s.d. than a single X-ray-derived structure. However, for the RNA-binding domain of nonstructural protein 1 of influenza A virus, consideration of an ensemble of conformations does not improve the agreement with the observed 13Calpha chemical shifts. Whether an ensemble of conformations rather than any single structure is a more accurate representation of a protein structure in the crystal as well as of the observed 13Calpha chemical shifts is determined by the dispersion of coordinates, in terms of the all-atom r.m.s.d. among the generated models; these generated models satisfy the experimental X-ray data with accuracy as good as the PDB structure. Therefore, generation of an ensemble is a necessary step to determine whether or not a single structure is sufficient for an accurate representation of both experimental X-ray data and observed 13Calpha chemical shifts in solution. PMID- 19564692 TI - A structural comparison of three isoforms of anionic trypsin from chum salmon (Oncorhynchus keta). AB - Three anionic salmon trypsin isoforms (CST-1, CST-2 and CST-3) were isolated from the pyloric caeca of chum salmon (Oncorhynchus keta). The order of catalytic efficiency (K(m)/k(cat)) of the isoforms during BAPA hydrolysis was CST-2 > CST-1 > CST-3. In order to find a structural rationalization for the observed difference in catalytic efficiency, the X-ray crystallographic structures of the three isoforms were compared in detail. Some structural differences were observed in the C-terminal alpha-helix, interdomain loop and active-site region. From the results of the detailed comparison, it appears that the structural flexibility of the C-terminal alpha-helix, which interacts with the N-terminal domain, and the substrate-binding pocket in CST-3 are lower than those in CST-1 and CST-2. In addition, the conformation of the catalytic triad (His57, Asp102 and Ser195) differs among the three isoforms. The imidazole N atom of His57 in CST-1 and CST 2 forms a hydrogen bond to the hydroxyl O atom of Ser195, but the distance between the imidazole N atom of His57 and the hydroxyl O atom of Ser195 in CST-3 is too great (3.8 A) for the formation of a hydrogen bond. Thus, the nucleophilicity of the hydroxyl group of Ser195 in CST-3 is weaker than that in CST-1 or CST-2. Furthermore, the electrostatic potential of the substrate-binding pocket in CST-2 is markedly lower than those in CST-1 and CST-3 owing to the negative charges of Asp150, Asp153 and Glu221B that arise from the long-range effect. These results may explain the higher catalytic efficiency of CST-2 compared with CST-1 and CST-3. PMID- 19564693 TI - Structures of mannose-6-phosphate isomerase from Salmonella typhimurium bound to metal atoms and substrate: implications for catalytic mechanism. AB - Mannose-6-phosphate isomerase (MPI) catalyzes the interconversion of mannose 6 phosphate and fructose 6-phosphate. X-ray crystal structures of MPI from Salmonella typhimurium in the apo form (with no metal bound) and in the holo form (with bound Zn2+) and two other structures with yttrium bound at an inhibitory site and complexed with Zn2+ and fructose 6-phosphate (F6P) were determined in order to gain insights into the structure and the isomerization mechanism. Isomerization involves acid/base catalysis with proton transfer between the C1 and C2 atoms of the substrate. His99, Lys132, His131 and Asp270 are close to the substrate and are likely to be the residues involved in proton transfer. The interactions observed at the active site suggest that the ring-opening step is probably catalyzed by His99 and Asp270. An active-site loop consisting of residues 130-133 undergoes conformational changes upon substrate binding. Zn2+ binding induces structural order in the loop consisting of residues 50-54. The metal atom appears to play a role in substrate binding and is probably also important for maintaining the architecture of the active site. Isomerization probably follows the previously suggested cis-enediol mechanism. PMID- 19564694 TI - The relationships among design experiments, invariant measurement scales, and domain theories. AB - In this paper we discuss principled design experiments, a rigorous, experimentally-oriented form of design-based research. We show the dependence of design experiments on invariant measurement scales. We discuss four kinds of invariance culminating in interpretive invariance, and how this in turn depends on increasingly adequate theories of a domain. These theories give an account of the dimensions and ordered attainments on a set of dimensions that span a domain appropriately. This account may be called a domain theory or learning theory of progressive attainments (in a local domain). We show the direct, and the broader benefits of developing and using these descriptive theories of a domain to guide prescriptive design approaches to research. In process of giving an account of this set of interdependencies, we will discuss aspects of the design method we are using, called Validity-Centered Design. This design framework guides the development of instruments based on domain theories, the development of learning opportunities; also based on domain theories, and the construction of a sound validity argument for systems that integrate learning with assessment. PMID- 19564695 TI - Considerations about expected a posteriori estimation in adaptive testing: adaptive a priori, adaptive correction for bias, and adaptive integration interval. AB - In a computerized adaptive test, we would like to obtain an acceptable precision of the proficiency level estimate using an optimal number of items. Unfortunately, decreasing the number of items is accompanied by a certain degree of bias when the true proficiency level differs significantly from the a priori estimate. The authors suggest that it is possible to reduced the bias, and even the standard error of the estimate, by applying to each provisional estimation one or a combination of the following strategies: adaptive correction for bias proposed by Bock and Mislevy (1982), adaptive a priori estimate, and adaptive integration interval. PMID- 19564696 TI - Local independence and residual covariance: a study of olympic figure skating ratings. AB - Rasch fit analysis has focused on tests of global fit and tests of the fit of individual parameter estimates. Critics have noted that slight, but pervasive, patterns of misfit to a Rasch model within the data may escape detection using these approaches. These patterns contradict the Rasch axiom of local independence, and so degrade measurement and may bias measures. Misfit to a Rasch model is captured in the observation residuals. Traces of pervasive, but faint, secondary dimensions within the observations may be identified using factor analytic techniques. To illustrate these techniques, the ratings awarded during the Pairs Figure Skating competition at the 2002 Winter Olympic Games are examined. The intention is to detect analytically the patterns of rater bias admitted publicly after the event. It is seen that the one-parameter-at-a-time fit statistics and differential item functioning approaches fail to detect the crucial misfit patterns. Factor analytic methods do. In fact, the competition was held in two stages. Factor analytic techniques already detect the rater bias after the first stage. This suggests that remedial rater retraining or other rater-related actions could be taken before the final ratings are collected. PMID- 19564697 TI - Constructing one scale to describe two statewide exams. AB - This study applies two approaches in creating a single scale from two separate statewide exams (Golden State Math Exam and California Standard Math Test) and compares some aspects of the two statewide tests. The first analysis involves a sequence of unidimensional Rasch scalings, using anchored items to scale the two tests together. The second analysis employs a 2-dimensional Rasch scaling using previous unidimensional analysis results to link the scales. The linking facilitates the investigation of their measurement properties of the two exams and is a basis for combining items from both exams to develop a more efficient testing program. The results of the comparisons of the two statewide exams based on the linking are shown and discussed. PMID- 19564698 TI - Multidimensional models in a developmental context. AB - The concept of epistemological development is useful in psychological assessment only insofar as instruments can be designed to measure it consistently, reliably, and without bias. In the psychosocial domain, most traditional stage assessment systems rely on a process of matching concepts in a scoring manual generated from a limited number of construction cases, and thus suffer to some extent from bias introduced by an over-dependence on particular content. On the other hand, Commons' Hierarchical Complexity Scoring System (HCSS) is an assessment that employs criteria for assessing the hierarchical complexity of texts that are independent of specific content. This paper examines whether the HCSS and one of the conventional systems, Kohlberg's Standard Issue Scoring System (SISS) measure the same dimension of performance. A multidimensional partial credit analysis was performed on data collected between 1955 and 1999. The correlation between performance estimates on the SISS and HCSS is 0.92. The high correlation provides strong evidence that the order of hierarchical complexity identified by the HCSS is the same latent dimension of ability assessed with the SISS. The HCSS produced more distinct patterns of ordered stages and wider gaps between adjacent stages. This evidence implies that individual performances display a higher degree of consistency in their hierarchical complexity under the HCSS. A developmental scoring system that employs scoring criteria that are independent of particular content might be more powerful than the traditional scoring systems as it provides easiness in scoring and also possibilities of cross-cultural, cross gender, cross-context comparison of conceptual knowledge within developmental levels. PMID- 19564699 TI - An application of the multidimensional random coefficients multinomial logit model to evaluating cognitive models of reasoning in genetics. AB - This article summarizes multidimensional Rasch analyses in which several alternative models of genetics reasoning are evaluated based on item response data from secondary students who participated in a genetics reasoning curriculum. The various depictions of genetics reasoning are compared by fitting several models to the item response data and comparing data-to-model fit at the model level between hierarchically nested models. We conclude that two two-dimensional models provide a substantively better depiction of student performance than does a unidimensional model or more complex three- and four-dimensional models. PMID- 19564700 TI - The ISR: intelligent student reports. AB - Rasch-based Scale Scores are a simple linear transformation of the basic logit metric. Scale Scores are the quantification of the measurement continuum. This quantification makes it possible to do arithmetic, computer differences, and apply standard statistical techniques. However, qualitative meaning is not in the numbers and must come from experience with the scale and from the descriptive information that can (and should) be attached. This includes item content and exemplars, normative information for relevant groups, historical data for the individual, and evaluative assessment like performance levels standards. The Scale Score metric is the structure that manages the organization of intelligent reports and recognizes anomalies. Scale Scores have no meaning, per se, but can provide a strong framework for organizing useful reports and presenting meaningful information. They facilitate diagnosis by "Analysis of Fit" and by "Analysis of Misfit." The Analysis of Fit relies on the general definition of the construct to describe what a student at a particular point on the scale can and cannot do. It is meaningful to the extent that the student conforms to the expectations of the measurement model. The Analysis of Misfit uses the model to identify surprises, i.e., departures from the model expectations. It highlights atypical areas of strong and weak performance. The intent is to bring these exceptons to the attention of the experts for informed, substantive interpretation and diagnosis. Intelligent reports, to be useful, and to justify the time and expense of testing, need to provide more information in a useable format than the candidate, student, parent, or educator had available otherwise. This requires more than reporting a single number or a single decision. It should include sufficient scaffolding to allow the consumer to extract quickly and efficiently all the useful information that can be taken from the test. Rasch Scale Scores are an important, perhaps essential, tool in this process. PMID- 19564701 TI - Obstructive sleep apnea and cardiovascular disease. AB - Over the past few decades, sleep apnea has emerged as an important potential etiologic factor in a broad range of cardiac and vascular diseases. These disease conditions include hypertension, coronary artery disease, myocardial infarction, heart failure, and stroke. Recognition of the role of sleep apnea in clinical cardiology is also increasing in Japan. Although sleep apnea has been strongly linked to obesity in Western populations, in Japanese and other Asian populations there is evidence to indicate that sleep apnea may be prevalent even at lower levels of obesity. In this review we address the epidemiology of sleep apnea. Since sleep apnea includes the combined stresses of hypoxemia, apnea, and disrupted sleep, we also review briefly the potential disease mechanisms that may be activated as a consequence of sleep apnea. We further examine the role of sleep apnea in the pathophysiology and management of specific cardiovascular conditions. Overall, while the evidence of sleep apnea as a causal mechanism in cardiovascular disease is strong and increasing, definitive evidence of the etiologic role of sleep apnea has yet to be obtained. The evidence is most clear in patients with hypertension. Also remaining to be established is whether the treatment of sleep apnea prevents cardiac and vascular events. With regard to this question, although the available data strongly suggest that continuous positive airway pressure treatment is beneficial, randomized control trials are needed in order to confirm this. PMID- 19564702 TI - Prehypertension and cardiovascular risk factor clustering among Mongolian population in rural and animal husbandry area, Inner Mongolia, China. AB - BACKGROUND: It is unclear whether and to what extent there is clustering of cardiovascular risk factors in the prehypertension phase among Mongolians in the rural and animal husbandry area of Inner Mongolia, China. METHODS AND RESULTS: The 2,589 Mongolian people aged > or=20 years served as subjects. Demographic data, lifestyle factors and family history of hypertension, blood pressure measurements, physical examination and blood samples were obtained and analyzed for all individuals. The proportions of 2 and > or =3 risk factors clustering were higher in hypertensives than in prehypertensives, and higher in prehypertensives than in normotensives (both P<0.01). After adjustment for age, sex and family history of hypertension, both prehypertension and hypertension were associated with mutually clustering of 5 risk factors; odds ratio (OR) of prehypertension with 1, 2 and > or =3 factors was 1.30 (1.02, 1.65), 1.93 (1.40, 2.67) and 2.44 (1.62, 3.68), respectively, and for hypertension the OR were 1.75 (1.31, 2.35), 3.84 (2.68, 5.48) and 6.95 (5.04, 10.63), respectively. CONCLUSIONS: There is clustering of risk factors in the phase of prehypertension among Mongolian people, so intervention measures should be taken to prevent progression to hypertension and other cardiovascular diseases. PMID- 19564703 TI - Branch segment occlusion with acute myocardial infarction is a risk for left ventricular free wall rupture. AB - BACKGROUND: Patients with acute myocardial infarction (AMI) whose culprit lesion lies in a branch of the 3 major coronary arteries have well-preserved cardiac function. A first MI with preserved cardiac function is a risk factor for left ventricular free wall rupture (LVFWR), so the aim of this study was to investigate the possible relationship between AMI with branch segment occlusion and LVFWR. METHODS AND RESULTS: The 439 patients with AMI were retrospectively studied. They were divided into 2 groups: group B (n=70; segments 4 atrioventricular node artery, 4 posterior descending coronary artery, 8, 9, 10, 12, 14, or 15 according to the AHA classification), and group P (n=369; segments 1, 2, 3, 5, 6, 7, 11, or 13). Primary percutaneous coronary intervention (PCI) was more often performed in group P (75% vs 57%; P=0.0018). In-hospital mortality tended to be lower in group B (1.4% vs 6.2%; P=0.105). The incidence of LVFWR was significantly higher in group B (10.0% vs 1.6%; P=0.0002).By multivariate logistic regression analysis, 1-vessel disease, absence of primary PCI, branch segment occlusion, and age were identified as independent predictors of LVFWR. CONCLUSIONS: The incidence of LVFWR was higher in group B and branch segment occlusion was identified as an independent predictor of LVFWR. PMID- 19564704 TI - Comparison of gene transcription between subcutaneous and visceral adipose tissue in Chinese adults. AB - Obese individuals with fat stored in visceral adipose tissue (VAT) generally suffer greater adverse metabolic consequences than those with fat stored predominantly in subcutaneous adipose tissue (SAT), but its molecular basis is not completely understood. We isolated paired samples of SAT and VAT from 15 lean and 15 obese subjects and systematically compared the transcription level of genes that may determine fat distribution and metabolic sequelae between SAT and VAT using quantitative real-time PCR. We found that, leptin levels were lower in VAT than SAT, for both lean and obese subjects. In lean subjects, tumor necrosis factor-alpha (TNF-alpha) was expressed equally in both fat depots, while toll like receptor 4 (TLR4) and glucocorticoid receptor (GR) showed significantly lower expression in VAT than SAT. In obese subjects, TNF-alpha and TLR4 expression were significantly higher in VAT than SAT, yet GR expression did not differ in these areas. For all subjects, VAT 11beta-hydroxysteroid dehydrogenate type 1 (11beta-HSD1) level was significantly correlated with BMI. GR expression level was significantly correlated with TLR4 expression level. Cultured adipocytes showed higher TLR4 mRNA level after differentiation, and higher TNF alpha level after treatment with free fatty acids. These results suggest that there are depot-specific differences in leptin, TNF-alpha, TLR4 and GR transcriptions in humans. TLR4 signaling and higher 11beta-HSD1 and GR levels in VAT may contribute predominantly to inflammatory factor production and subsequent metabolic sequelae in obese human. PMID- 19564705 TI - Unusual clinical and pathological presentation of a neuroendocrine tumor in a patient with multiple endocrine neoplasia type 1. AB - Neuroendocrine tumors develop in various organs in patients with multiple endocrine neoplasia type 1 (MEN1). Among those, tumors developed in upper gastrointestinal tract, thymus and bronchus have historically been called "carcinoid tumor". Occurrence of "carcinoid tumor" in other region is very rare and molecular pathogenesis of such tumors is unknown. We have experienced a patient with MEN1 who have developed an "ectopic" retroperitoneal neuroendocrine tumor. Genetic analysis of the MEN1 gene in tumor cells revealed a somatic mutation in exon 9 as well as a germline mutation in exon 10. Allele-specific amplification followed by sequence analysis revealed these two mutations exist on the different allele, indicating both alleles are functionally inactivated. Immunohistochemical staining with an anti-menin antibody revealed that wild-type menin is not expressed in tumor cells. Expression of p27(Kip1) protein is not observed in tumor cells, either. These results confirmed the inactivation of the MEN1 gene as a genetic cause of an ectopically developed neuroendocrine tumor in a patient with MEN1. PMID- 19564706 TI - Additional beneficial effects of recombinant growth hormone in alendronate treated patients with idiopathic osteoporosis. AB - In order to study the benefit of adding recombinant human growth hormone (rhGH) to antiresorptive therapy, six patients with idiopathic osteoporosis (IO) receiving alendronate plus calcium and vitamin D were started on daily subcutaneous injections of rhGH 2.0 IU for one year. Fasting morning urine and serum samples were collected for N telopeptide of type-1 collagen (NTX), serum bone-specific alkaline phosphatase (BSAP) and insulin-like growth factor 1 (IGF 1) during the study. Bone mineral density (BMD) was determined by dual-energy x ray absorptiometry at baseline and 01 year. The effect of rhGH was evaluated comparing the percentage changes in BMD during the last year on ALN with the results obtained with the combined therapy. Serum IGF-1 increased in all patients but variations were not significant (p=0.266). Serum BSAP did not significantly change (p=0.078) but median NTX increased at 45 days from 12.3 to 19.8 nMBCE/mMCr (p=0.012) and tended to return to baseline values at 12 months (15.2 nMBCE/mMCr). Comparing with isolated ALN therapy, a beneficial effect on bone density was observed in 2/3 of the patients at lumbar spine, and percentage change (median and quartiles) varied from -0.65% (-2.33 and 2.23) on ALN to 0.70% (-0.35 and 3.03) on ALN+GH. Although no bone gain occurred at the femoral neck, our data point to a positive effect of rhGH in patients with idiopathic osteoporosis. PMID- 19564707 TI - Adrenal Cushing's syndrome due to bilateral macronodular adrenal hyperplasia: prediction of the efficacy of beta-blockade therapy and interest of unilateral adrenalectomy. AB - Bilateral adrenalectomy is the standard treatment for Cushing's syndrome (CS) related to ACTH-independent bilateral macronodular hyperplasia (AIMAH), although it imposes life-long adrenal insufficiency. This study reports a clinical case in order to discuss the clinical interest of pharmacological beta-blockade of illegitimate membrane receptors and unilateral adrenalectomy as alternatives to bilateral adrenalectomy for treatment of CS due to AIMAH. Evidence for cortisol stimulation by upright posture and insulin-induced hypoglycemia in a patient with CS related to AIMAH led us to try beta-blockers as a therapeutic test and then as a first line treatment. Thus, a 3-day beta-blocker test (320 mg/d propranolol) induced normalization of cortisol secretion, with return of hypercortisolism at the end of the test. A long term treatment with 320 mg/d propranolol allowed sustained normalization of cortisol secretion and progressive disappearance of Cushingoid features but after 8 months the patient complained of Raynaud's syndrome and fatigue. Lowering propranolol dosage or switching to atenolol was less efficient to reduce cortisol levels. Unilateral adrenalectomy was then performed as a second line treatment, leading to normalisation of the 24h urinary cortisol without adrenal insufficiency. Long term control of blood pressure and glycemia were observed during a 7-year follow-up without beta-blocker. In conclusion, a 3-day propranolol test may identify patients with AIMAH who can benefit from a long term beta-blocker treatment. In case of intolerance to beta blocking agents, unilateral adrenalectomy may allow for long term control of Cushing's syndrome related to AIMAH without adrenal insufficiency. PMID- 19564709 TI - [Regulatory B cells]. AB - B cells positively regulate immune responses through antibody production and optimal CD4(+) T cell activation. However, a specific and functionally important subset of B cells can also negatively regulate immune responses in mouse autoimmunity and inflammation models. The lack or loss of regulatory B cells has been demonstrated by exacerbated symptoms in experimental autoimmune encephalitis, chronic colitis, contact hypersensitivity, collagen-induced arthritis, and non-obese diabetic mouse models. We have recently found that IL-10 producing regulatory B cells predominantly localize within a rare CD1d(hi)CD5(+) B cell subset that shares cell surface markers with both B-1 and marginal zone B cells. We have labeled this specific subset of regulatory B cells as B10 cells to highlight that these rare CD1d(hi)CD5(+) B cells only produce IL-10 and are responsible for most IL-10 production by B cells, and to distinguish them from other regulatory B cell subsets that may also exist. This review focuses on the recent progress in this field and the exciting opportunities for understanding how this unique B cell subset influences diverse immune functions. PMID- 19564710 TI - [IgA subclass and IgA deficiency]. AB - There are two subclasses of IgA, IgA1 and IgA2, and its heavy chains are encoded by two different genes, alpha1 and alpha2 genes. These two subclasses play important roles in the first line of defense, and the amount ratio of these molecules in secretions varies. IgA deficiency (IgAD) is the most common immunodeficiency, however the pathogenesis in most cases of IgAD is unknown. The class switch disorder in IgA producing B lymphocytes is one of the important factors in IgAD patients. The decreased expression levels of Ialpha germline transcripts before a class switch may be the cause of selective IgAD. The alpha1 and alpha2 gene expression levels are low in most IgAD patients. Using RT-PCR method in which alpha1 and alpha2 mRNAs can be separately evaluated, we identified the second case of alpha1 gene deletion in Japan. Longitudinal change in the serum IgA of the patient with alpha1 gene deletion showed the pattern of the partial IgAD. Patients with alpha1 gene deletion can be considered as having partial IgAD. PMID- 19564711 TI - [New biologic and non biologic disease modifying anti-rheumatic drugs for rheumatoid arthritis]. AB - TNF inhibitors, infliximab, etanercept and adalimumab, and tocilizumab are available in Japan and have been successful at improving the signs and symptoms of RA and, thereby, have set a new standard for disease control of RA and have the potential to protect joints from structural damage or to improve quality of life and mortality. However, the rate of successful induction of remission was about 30% and the treatment strategies for the patients who do not respond to these biologics should be established. Randomized clinical trials targeting T or B lymphocytes have been conducted in addition to the new anti-TNF blockers like golimumab or certolizumab pegol. Anti-CD20 antibodies such as rituximab (chimeric), ocrelizumab (humanized), ofatuzumab (full human) demonstrated effectiveness to the patients who do not respond to TNF blockers. CTLA-4 Ig, which can transduce negative signal into T lymphocytes in the co-stimulatory pathway, has also showed a good response to refractory RA. Furthermore, low molecular agents such as Jak (Janus kinase) 3 or syk (spleen tyrosine kinase) inhibitors demonstrated rapid and strong suppression of synovitis and are thought to be new candidates for the drugs to overcome refractory RA. PMID- 19564712 TI - [Molecular mechanisms of tissue fibrosis]. AB - Tissue fibrosis is a common cause of organ failure. Consequently, elucidation of the mechanisms underlying both the initiation and progression of fibrosis is an essential step toward establishing new therapeutic strategies for the treatment of organ failure. Fibroblasts are the principal effectors mediating fibrosis and their heterogeneous origins, including epithelial-mesenchymal transition (EMT), bone marrow-derived cell or fibrocyte, and endothelial-mesenchymal transition (EndMT), have been demonstrated. Chronic hypoxia has been proposed as an important microenvironmental factor in the development of tissue fibrosis. Recently, we reported that hypoxia induces EMT in renal tubular epithelial cells through activation of hypoxia-inducible factor-1alpha (HIF-1alpha). Using the Cre loxP mediated gene targeting of HIF-1alpha or VHL which acts as a ubiquitin ligase to promote degradation of HIF-1alpha, we showed that HIF-1alpha plays a key role in the progression of renal fibrosis. As a large number of molecules that contribute to the induction of fibrosis have been identified, and their signal transduction pathway has been characterized, these fibrosis-related molecules have been proposed as therapeutic targets. EMT antagonists, TGF-beta signal modulator, and HIF-1alpha inhibitor could be useful for the treatment of fibrosis. PMID- 19564713 TI - [Biologics in current therapy for inflammatory bowel disease]. AB - Recent advance of molecular biology and immunology contributes to the development biologics such as anti-TNFalpha agent in the treatment for inflammatory bowel disease (IBD). Although therapeutic strategy of IBD has not been changed for a long time, success of Infliximab, first anti-TNFalpha agent, now changes therapeutic strategy of IBD dramatically. Top-down strategy has been considered to improve patients' natural history in the therapy for Crohn's disease as well as in rheumatoid arthritis. Infliximab also has been expected as a promising medicine for pediatric Crohn's disease. Furthermore, Infliximab has been approved for the therapy of ulcerative colitis. These tremendous successes of Infliximab have encouraged us to develop other anti-TNF agents and other biologics. In this review, we describe current topics of biologics in IBD treatment and discuss future direction. PMID- 19564714 TI - Elevation of plasma eotaxin levels in children with food allergy. AB - BACKGROUND: Eosinophils play an important role in allergic responses. Eotaxin is a CC chemokine that promotes the selective recruitment of eosinophils. This study was performed to investigate the significance of eotaxin in pediatric food allergies. METHODS: The study population included 35 patients with food allergy, 18 patients with atopic dermatitis but without food allergy, and 19 age-matched non-allergic controls. Eotaxin-1 and eotaxin-3 levels in plasma were assayed by enzyme-linked immunosorbent assay. Simultaneously, eosinophil counts in peripheral blood and serum immunoglobulin E (IgE) values were assessed. RESULTS: Plasma eotaxin-1 levels were 93.6+/-33.4 pg/ml in patients with food allergy, 78.0+/-31.8 pg/ml in patients with atopic dermatitis, and 60.4+/-15.7 pg/ml in controls. Differences between the food allergy and control groups were significant (P<0.001). Circulating eosinophil counts in patients with food allergy were higher than those in controls (5.84+/-9.46x10(9)/l vs. 1.20+/ 1.11x10(9)/l, P<0.001). Nevertheless, eotaxin-1 levels in children with food allergy were not correlated with eosinophil counts or serum IgE levels. There were no significant differences in eotaxin-3 levels between the 3 groups. CONCLUSION: Plasma levels of eotaxin-1 were elevated in children with food allergy. The pathophysiological relevance of the increase in eotaxin is discussed. PMID- 19564715 TI - Bone erosion of the sternocostal joint in a patient with Behcet's disease. AB - Behcet's disease (BD) is a polysymptomatic and recurrent systemic vasculitis with a chronic course and unknown cause. Erosive arthropathy is extremely rare. We report a 52-year-old female patient with BD demonstrating bone erosion of the sternocostal joint. PMID- 19564716 TI - [A case of microscopic polyangiitis relapsed with diffuse alveolar hemorrhage and rapidly progressive glomerulonephritis]. AB - The patient was a 63-year-old woman. She was admitted to our hospital with acute renal failure and multiple mononeuritis in 2002. She was diagnosed as microscopic polyangiitis based on positive for MPO-ANCA. Remission was induced by combination therapy with methylprednisolone pulse therapy and plasma exchange. Because condition of the disease was stable, prednisolone was discontinued from August 2006. Elevation of serum creatinine and microscopic hematuria was detected in November 2007. Fever and dyspnea occurred in January 24 2008. Elevation of CRP and serum creatinine was found, and infiltration in bilateral lung was noted on chest X-ray. She was admitted on the same day. After admission, she presented with hemosputum and exacerbation of dyspnea. Chest CT revealed diffuse consolidation and ground glass opacity, and MPO-ANCA converted to be positive. Diagnosis of diffuse alveolar hemorrhage and rapidly progressive glomerulonephritis with microscopic polyangiitis was made, and she was managed by artificial respirator and CHDF in ICU. Combination therapy with steroid pulse therapy and plasma exchange re-induced remission. Mizoribine was administrated as maintenance therapy with oral prednisolone. PMID- 19564717 TI - [Clinical and histological experience of labial lip biopsy in juvenile Sjogren syndrome]. AB - The therapeutic efficacy of corticosteroids and immunosuppressants on secretary glands of children with Sjogren syndrome was investigated examining the lip biopsy specimen on both lymphocyte infiltration and fibrosis. Six children with primary Sjogren syndrome and two children with lupus-associated secondary Sjogren syndrome were evaluated according to the intensity of therapy. The shorter the term of medications of corticosteroids and immunosuppressants were, the lesser the extents of lymphocyte infiltration and fibrosis were, and the lower the doses of medications were, the lesser the efficacy was. Thus, in childhood Sjogren syndrome, appropriate corticosteroids and immunosuppressants may provide the suppressive effects on the progressive inflammatory destruction of secretary glands. Further evaluation with more patients is needed to determine the inclusion criteria of these treatments for sicca syndrome, especially in cases with no other organ involvement. PMID- 19564718 TI - Current iodine status in Turkey. AB - OBJECTIVES: To evaluate the current nationwide iodine status in Turkey by determining urinary iodine concentrations (UIC) and household salt iodine content. A follow- up monitoring study was also conducted in 30 urban areas. METHODS: A school-based survey was conducted in 2007 by using multistage 'proportionate to population size' (PPS) cluster sampling method. The study population was composed of 900 school-age children (SAC) from different urban, suburban, and rural areas. UIC and iodine content of the table salt used at home were analyzed. RESULTS: Median UIC was 107 microg/l (147 in urban, 42 in suburban and rural areas, p<0.001). There were severe iodine deficiency (ID) in 7.2%, moderate and mild ID in 20.6% and 19.3%, of the SAC, respectively. UIC was sufficient (>100 microg/l) in 50% of the study population, whereas it was excessive (>300 microg/l) in 10.5% of them. Of the 900 salt samples, 662 (73.5%) were iodized and 508 samples (56.5%) contained adequately iodized salt (iodine content >15 ppm). UIC of the study population and salt iodine levels correlated well (r=0.42, p<0.001). CONCLUSIONS: Moderate to severe ID still exists in 27.8% of the Turkish population, which is much better compared to 1997 and 2002 surveys (i.e. 58%, 38.9%, respectively). The follow-up monitoring study (in 2007) demonstrated that ID has been eliminated in 20 of 30 cities surveyed, and median UIC was 130 microg/l. ID has been eliminated in most of the urban population, however, it is still an important problem in rural areas and in particular geographical regions, which should be the target of future programs. PMID- 19564719 TI - Cardiac remodeling in patients with primary aldosteronism. AB - OBJECTIVE: To evaluate the morpho-functional changes of the myocardium in patients with primary aldosteronism (PA). DESIGN: An observational study in a university referral center for blood pressure diseases. PATIENTS: Twenty- three patients with PA, 24 patients with essential hypertension (EH), and 15 normotensive controls (C) underwent conventional echocardiography with integrated backscatter (IBS) and tissue Doppler imaging (TDI) analysis. The corrected IBS (C IBS) values and the systo-diastolic variation of IBS (CV-IBS) were performed at both interventricular septum and the posterior wall levels. TDI myocardial systolic (Sm), early diastolic (Em), and late diastolic (Am) velocities of both left ventricular walls were also determined. RESULTS: In PA patients, septal and posterior wall CV-IBS were significantly lower than C (p<0.0001) and EH patients (p<0.001). In EH, CV-IBS was significantly lower than C (p<0.001). Patients with PA exhibited lower Sm, lower Em, and higher Am, and a subsequently reduced Em/Am ratio than C (p<0.001 for all) and EH (p<0.01 for all) at interventricular septum and lateral wall levels. In the latter, Sm, Em, and Em/Am ratio were lower and Am was higher than C (p<0.001 for all). In PA and EH patients, CV-IBS at both septum (r=-0.66, p<0.001) and posterior wall levels (r=-0.67, p<0.001) and Sm peak of both septum (r=-0.52, p<0.001) and lateral wall (r=-0.55, p<0.001) were inversely related to plasma aldosterone. CONCLUSIONS: Patients with PA showed myocardial wall remodeling characterized by increased myocardial fibrosis and early left ventricular systodiastolic function abnormalities. PMID- 19564720 TI - Growth hormone response to physical exercise in growing patients with classic congenital adrenal hyperplasia. AB - Glucocorticoid over-treatment in children with congenital adrenal hyperplasia (CAH) may suppress GH secretion and growth. Aims of our study were: 1) to evaluate post-exercise GH response in patients affected by CAH due to 21 hydroxylase deficiency, in comparison with a group of healthy subjects; 2) to investigate the relationship between the hormonal markers of adequate steroid therapy and GH secretion. We evaluated GH secretion every 6 months in 20 young CAH patients (8 girls, 12 boys). Mean follow-up was 4.6+/-0.9 yr (107 tests performed, 5.35+/-2.05 repeated tests for each patient). Forty-four healthy subjects (25 boys, 19 girls) were selected as a control group. The range of post exercise GH peak was very wide, but medians were not statistically different in cases and controls (p=0.570). Multivariate analysis showed that post-exercise GH peak was not related to age (p=0.743), gender (p=0.296) or pubertal status (p=0.440) in both groups. GH increase from baseline showed the same behavior (p=0.265, 0.639 and 0.105, respectively). In CAH patients, GH peak and GH increase were both directly related to 17-OH-progesterone levels [GH peak: p=0.032--95% confidence interval (CI): 0.01-0.34--beta=0.18; GH increase: p=0.008 -95% CI: 0.06-0.35--beta=0.20]. The negative effect of glucocorticoid therapy on GH secretion seems to be dominant in CAH. The most effective approach to adjust treatment remains monitoring growth. Relying on hormonal markers to adequate steroid therapy may result in over-treatment, GH suppression, and finally poor linear growth. PMID- 19564721 TI - Severe bone disease with bilateral femur fracture in a young woman after gastrectomy for gastric polyposis with SMAD4 mutation. PMID- 19564722 TI - Increased ratio of mRNA expression of the genes CYP17 and CYP11B1 indicates autonomous cortisol production in adrenocortical tumors. AB - OBJECTIVE: Due to increased use of imaging techniques, adrenal incidentalomas are frequently detected. The majority are non-hyperfunctioning adrenocortical tumors. We have previously shown that expression of the gene CYP17, coding for the enzyme in the cortisol pathway, correlates with cortisol release from adrenocortical tumors in vitro. The aim of this study was to compare clinical data with mRNA expression of CYP17 and CYP11B1 in adrenocortical tumors from patients with and without Cushing's syndrome and to identify adrenal tumors that may cause subclinical Cushing's syndrome. DESIGN: A retrospective study of 34 patients undergoing adrenalectomy due to an adrenal tumor. METHODS: Clinical data were collected. In the adrenal gland the mRNA expression of the genes CYP17 and CYP11B1 was studied with in situ hybridisation technique. RESULTS: The median ratio of CYP17/CYP11B1 expression in tumors from patients with Cushing's syndrome was significantly higher than the median ratio in the non-hyperfunctioning tumors. Tumors from 2 patients with subclinical Cushing's syndrome had ratios within the upper range for non-hyperfunctioning tumors. CONCLUSIONS: The ratio between the expression of the genes CYP17 and CYP11B1 in tumors from patients with Cushing's syndrome is significantly higher than in the non-hyperfunctioning tumors. This indicates that 17alpha-hydroxylase is a major determinant of cortisol overproduction. The patients with subclinical Cushing's syndrome in this study are too few to draw any firm conclusions although the results suggest that subclinical Cushing's syndrome may be identified post-operatively with this method. PMID- 19564723 TI - Detectable cerebrospinal fluid HIV RNA with associated neurological deficits, despite suppression of HIV replication in the plasma compartment. PMID- 19564724 TI - Are the effects of isoniazid preventive therapy and highly active antiretroviral therapy additive in preventing HIV-associated tuberculosis? PMID- 19564725 TI - Additive effects of isoniazid preventive therapy and HAART. PMID- 19564726 TI - Idiopathic central pontine myelinolysis in an eunatremic patient with AIDS. PMID- 19564727 TI - Disruption of an env tyrosine-dependent sorting signal does not affect susceptibility of HIV-1 to cytotoxic T lymphocytes. PMID- 19564733 TI - Bibliography. Current world literature. PMID- 19564738 TI - Safety of general anesthesia for lumbar puncture and bone marrow aspirate/biopsy in pediatric oncology patients. AB - INTRODUCTION: Painful short duration procedures such as bone marrow aspiration/biopsy and the lumbar puncture with or without intrathecal chemotherapy are frequently performed during the treatment of children with cancer. This study evaluated the frequency and severity of complications of bone marrow aspiration biopsy and lumbar puncture/intrathecal chemotherapy under general anesthesia. PATIENTS AND METHODS: A prospective observational study was performed from November 2003 to August 2005. Patients with cancer younger than 21 years old, receiving treatment at the Pediatric Oncology Unity of Hospital de Clinicas de Porto Alegre, undergoing diagnostic and/or therapeutic short duration procedures carried out under general anesthesia in the outpatient surgery unit. RESULTS: One hundred and thirty-seven patients were submitted to 423 procedures under general anesthesia. There were 61% boys, mean age of 7.5 years (0.2 to 21) and ASA II 98%. Eighty seven percent of the procedures were carried out in patients with leukemia or lymphoma. The majority of the procedures had no adverse events during intraoperative and postoperative periods. No procedure had to be suspended after it had begun. One patient had lumbar pain after the procedure and was admitted to the ward with suspected subdural bleeding, but this was not confirmed. No patient needed cardiopulmonary reanimation or treatment in the intensive care unit. CONCLUSIONS: General anesthesia for short duration painful procedures in children undergoing treatment for malignancies is safe when carried out by trained professionals in outpatient clinical surgery unit. PMID- 19564739 TI - Neuroblastoma in Turkish children: experience of a single center. AB - OBJECTIVE AND METHOD: The survival of the patients with neuroblastoma has improved in last few decades. But still it depends on various clinical and biological factors. To assess the clinical features and trends in survival, the data for 500 newly diagnosed patients between January 1972 and December 2004 from a single center were retrospectively analyzed. RESULTS: Histopathologic subtypes were neuroblastoma (NBL) in 462 patients (92.4%) and ganglioneuroblastoma in 38 patients (7.6%). The median age was 2.9 years and Male/Female ratio was 1.3/1. Primary tumor sites were abdomen, thorax, pelvis, neck, and others with the frequency of 72.2%, 14.9%, 3.8%, 3.2%, and 5.9%, respectively. There were 30, 49, 133, 257, 31 patients with stage 1, 2, 3, 4, 4S disease and their 10-year survival rates were 100%, 75.8%, 34.1%, 6.5%, and 59.4%, respectively. The outcome has significantly improved according to 10-year periods. The 5-year overall survival rates were 14%, 26.1%, 39.2%, and 52.4% for the years of 1970s, 1980s, 1990s, and after 2000. Surgical procedure involving total or near total tumor removal improved the survival (P=0.002). Both 5-year overall survival and event free survival rates were higher when partial resection was performed, especially in stage 3 disease (P=0.002 and P=0.02). In multivariate analysis, age above 18 months at diagnosis (P=0.01), stage 4 disease (P<0.001), abdominal primary tumor site (P<0.001), NBL subtype in histopathology (P=0.001), responsiveness to chemotherapy (P<0.001) positive or high Vanillyl mandelic acid levels (P=0.02) and male sex (P=0.008) were the determinants of poor prognosis. CONCLUSIONS: The survival rates in children with local disease are comparable with the results of developed countries; however, the results in children with advanced disease are still not satisfactory. To improve the outcome, especially in children with advanced disease, more effective chemotherapy regimens and molecular therapies should be investigated. Sharing the knowledge and capacity building to improve the treatment results in NBL are also critical for developing countries. PMID- 19564740 TI - Prognostic factors in pediatric cancer patients admitted to the pediatric intensive care unit. AB - Higher mortality and morbidity are well established in children with malignancies in whom intensive care admissions are required. A retrospective cohort study was conducted to assess the risk factors for children with cancer in the pediatric intensive care unit (PICU) for short-term outcome (survival vs. nonsurvival when leaving the PICU). The records of 36 children with a median age of 5 years (range: 0.5 to 21) between August 2004 and August 2007 were reviewed. Mortality rate was 55%, higher than the yearly overall PICU mortality rate of 12% (P<0.0001). The mean Pediatric Risk of Mortality Score (PRISM) III score among survivors was lower than that among nonsurvivors (9.4+/-5.7 vs. 16.4+/-5.3, P=0.001). Comparison of observed and predicted mortality derived from the PRISM III score showed that distribution of outcome was not different and the prediction model performed well (goodness of fit test: chi=3.64, df=6, P=0.725). The mortality rates were 66.6% and 33.3% in patients with high (>10 points) and low (< or =10 points) PRISM III score, respectively (P=0.05). Mortality rate was significantly related to presence and number of organ system dysfunction (P=0.031 and P=0.013, respectively), sepsis (P=0.05), mechanical ventilation (P=0.005), and positive inotropic support (P=0.003). By using multiple logistic regressions, the independent risk factor was PRISM III score at the time of admission to PICU (P=0.05). The PRISM III score performed well as a predictor of outcome. For decision to admit such patients to the PICU or to forgo life-sustaining therapies, other factors such as need for mechanical ventilation and positive inotropic support, presence and numbers of organ system dysfunction should be taken into consideration as well. PMID- 19564741 TI - Postoperative fevers in pediatric solid tumor patients: how should they be managed? AB - It is unclear how aggressively postoperative fevers should be managed in immunosuppressed pediatric oncology patients after major surgery. Little data exists on this subject. Therefore, a retrospective study of patients treated at our center was undertaken to examine this. PURPOSES: (1) to describe the prevalence of fever and infection in postoperative pediatric solid tumor patients undergoing primary tumor resection, (2) to examine the risk factors that contribute to the development of postoperative infections, and (3) to describe the variation in practice in managing fevers. METHODS: Chart reviews were performed on patients diagnosed with a spectrum of tumor types from January 2000 to October 2005 who received preoperative chemotherapy, followed by tumor resection. RESULTS: Ninety-eight children met inclusion criteria and 73 (74%) developed fevers postoperatively; 14% of these had documented infections and 1 patient died from sepsis. Factors associated with increased risk of infection were a diagnosis of neuroblastoma (P=0.015), and surgery longer than 8 hours (P=0.059). The investigation and management of postoperative fevers varied in these patients. CONCLUSIONS: Postoperative fevers may be indicative of severe infection. We suggest that a standardized approach to the management of these patients, including prompt physical assessment, clinical investigations, and empiric antibiotic consideration is vital to minimize complications. PMID- 19564742 TI - Survey of the long-term use of peripherally inserted central venous catheters in children with cancer: experience in a developing country. AB - Peripherally inserted central catheters (PICCs) have been used for many years in developed countries, but few studies have been focused on children with cancer in developing countries. In this study, we assessed the feasibility of PICCs and determined the rate of PICC-related complications in children with cancer. We prospectively followed all children with cancer over 3 years of age who received chemotherapy and PICC placement in our cancer center between June 2003 and May 2007. The date of last follow-up was January 31, 2008. A total of 119 PICCs were inserted into 116 patients during the 48-month period. PICCs were placed in 113 of 119 attempts, yielding an insertion rate of 95.0%. The 113 PICCs were in place for a total 26,721 catheter days (median time, 246 d; range, 8 to 455 d). The 113 PICCs had 53 overall complications, for a rate of 1.98/1000 catheter days. Twenty one (18.6%) PICCs were removed because of complication with a rate of 0.79/1000 catheter days. The most common reason for PICC removal was breakage/leakage. An infection requiring PICC removal occurred in 4 patients. This study demonstrated relatively low complication rate and long duration for PICCs in children with cancer over 3 years of age in our hospital. PMID- 19564743 TI - P-glycoprotein activity predicts outcome in childhood acute lymphoblastic leukemia. AB - Treatment of children with acute lymphoblastic leukemia (ALL) is based on P glycoprotein (P-gp)-dependent cytostatics. We assessed the P-gp function in blast cells as a possible prognostic factor and its influence on the overall survival. P-gp function was measured using the verapamil-sensitive Rhodamine efflux. Cell samples from 7 of 45 (16%) patients revealed rhodamine-efflux positive blasts. There were no relations between the presence of P-gp, clinical characteristics (age, sex, hepatomegaly, and splenomegaly) and initial laboratory parameters (immunophenotype, white blood cells count, and serum lactate dehydrogenase) in ALL. P-gp activity plays a negative role, both for a remission achieved on day 33 and for susceptibility to steroid therapy. Children bearing rhodamine-efflux positive blasts had a significantly shorter 5-year overall survival of 35%, as compared with 74% in those negative for P-gp function. Lack of any association with clinical characteristic and initial laboratory parameters suggests that presence of P-gp is an independent prognostic factor. PMID- 19564744 TI - Pediatric oncology in pakistan experiences of a visiting "senior expert". PMID- 19564745 TI - Ganglioneuroblastoma-associated vitamin D deficiency rickets. AB - Vitamin D deficiency is the most common cause of rickets mainly in breast-fed dark-skinned, African or Asian children receiving inadequate sunlight exposure. We report a case of a 1.5 year-old Afro-Italian male infant living in South Italy who came to our observation with the typical clinical picture of vitamin D deficiency rickets. The child was exclusively breast-fed for 8 months without vitamin D supplements. Owing to the rarity of vitamin D deficiency rickets in the South of Italy he underwent several investigations, which demonstrated the association with an abdominal ganglioneuroblastoma. To our knowledge, ganglioneuroblastoma has never been reported in association with vitamin D deficiency rickets. Although the association between these 2 rare conditions may be coincidental, the protective action of vitamin D against cancer suggests that vitamin D deficiency might have contributed to the development of ganglioneuroblastoma in our patient. PMID- 19564746 TI - Posterior reversible encephalopathy syndrome in childhood with hematologic/oncologic diseases. AB - Posterior reversible encephalopathy syndrome (PRES) is a clinico-neuroradiologic disease entity represented by characteristic magnetic resonance image (MRI) findings of subcortical/cortical hyperintensity in T2-weighted sequences, more often observed in parieto-occipital lobes, accompanied by clinical neurologic alterations. PRES is a rare central nervous system complication in childhood hematologic-oncologic patients and shows very different neurologic symptoms between patients, from numbness on extremities to generalized seizure. The etiology of PRES was not well known until these days. In this study, 8 patients with PRES were reviewed, retrospectively. There were 4 patients with acute lymphocytic leukemia, 1 with aplastic anemia, and 3 with solid tumors (1 patient each for neuroblastoma, Ewing sarcoma, and osteosarcoma). Allogeneic stem cell transplantation was performed in 2 patients. Immunosuppressive agents such as tacrolimus and cyclosporine A were used in 3 patients. One neuroblastoma patient was in immediate postoperative status. All patients experienced seizure attacks of different types and showed typical MRI findings. Follow-up MRIs revealed significant improvements. From this review, we might consider chemotherapy and surgery as additive causes for PRES other than immunosuppressive agents. Therefore, careful examination of the patients receiving chemotherapy and surgery was needed to find out this uncommon but good prognostic complication. PMID- 19564747 TI - Oral eruption cysts in a child with hepatoblastoma. AB - Oral eruption cysts develop when extravasated fluid, epithelial remnants of tooth embryogenesis, and blood obliterates the submucosal space encapsulating an erupting primary or permanent tooth. In immunocompetent children, these lesions are treated conservatively with watchful monitoring for spontaneous rupture, mucosal healing, and timely tooth emergence. We describe the clinical course for 2 oral eruption cysts in a child with Stage III hepatoblastoma treated with chemotherapy before liver transplant. This article provides recommendations for care when prophylactic surgical excision of oral eruption cysts is indicated in pediatric oncology patients. PMID- 19564748 TI - Better posttransplant outcome with fludarabine based conditioning in multitransfused fanconi anemia patients who underwent peripheral blood stem cell transplantation. AB - Several investigators have been looking for less toxic conditioning regimen for stem cell transplantation in Fanconi anemia (FA) patients because of sensitivity to DNA cross-linking agents and tendency to malignancy. We report 16 multitransfused FA patients who underwent peripheral stem cell transplantation from 13 related and 3 unrelated donors. Although the first 6 patients received thoraco-abdominal irradiation + cyclophosphamide + antithymocyte globulin (regimen A) for conditioning, fludarabine (FLU) + cyclophosphamide + antithymocyte globulin (regimen B) were used in the last 10 patients in which 3 of them received unrelated graft. Cyclosporin A was given alone for the related allografts but also included mycophenolate mofetil for the unrelated allograft as graft versus host disease prophylaxis. We observed a lower risk of peritransplant morbidity and mortality with fewer and milder graft versus host disease in FLU based group. We lost 3 patients in regimen A group and 1 of them from secondary acute myeloid leukemia. Three patients are alive with transfusion independent. In regimen B group, 9 of 10 patients are alive with normal hematologic parameters and full donor chimerism. The longest follow-up durations are 90 and 60 months in regimen A and B, respectively. In conclusion, FLU based conditioning is more effective and successful with lower toxicity in multitransfused FA patients. However, it needs more experience and longer follow up duration. PMID- 19564749 TI - Management of neonatal aortic arch thrombosis with low-molecular weight heparin: a case series. AB - Aortic arch thrombosis (AAT) is a potentially life threatening condition in neonates. We report 3 neonates with AAT and complete occlusion of a unilateral internal carotid artery with multiple cerebral infarcts. Multidisciplinary input led to deferral of thrombolytic therapy and surgical thrombectomy owing to the potential risk of hemorrhage within the infarcts. All the neonates were treated with low-molecular weight heparin treatment alone without any complications. Thrombus resolution was complete in 2 neonates, whereas it was near complete in one neonate. All the infants showed normal growth and development during follow up. The rewarding outcome in these cases suggests that treatment with low molecular weight heparin could be considered an option to treat AAT in selected patients. PMID- 19564750 TI - Kasabach-merritt phenomenon: case series and retrospective review of the mayo clinic experience. AB - Kasabach-Merritt phenomenon (KMP) is a rare thrombocytopenic consumption coagulopathy associated with an enlarging tufted angioma or kaposiform hemangioendothelioma. We report a case series of patients with KMP and discuss the treatment options and outcomes. Nine patients with a diagnosis of KMP were identified through retrospective chart review-6 had "definite KMP" and 3 had "less likely KMP." The hematologic features of KMP and those of chronic coagulopathy seen with other vascular malformations can be similar, which makes KMP difficult to distinguish. PMID- 19564751 TI - Hypoplastic glomerulocystic kidney disease and hepatoblastoma: a potential association not caused by mutations in hepatocyte nuclear factor 1beta. AB - Hypoplastic glomerulocystic kidney disease is an autosomal dominant disorder caused by mutations in hepatocyte nuclear factor-1beta. Hepatoblastoma is a sporadic occurring tumor of embryonal origin that has been associated with the several overgrowth syndromes. We report a case of concomitant hypoplastic glomerulocystic kidney disease and hepatoblastoma. Review of the literature identified 4 other patients with a similar association. We propose that hypoplastic glomerulocystic kidney disease and hepatoblastoma represent a possible association, and we excluded mutations in hepatocyte nuclear factor 1beta in our patient as causative of this putative association. PMID- 19564752 TI - Multiple neoplasia in a 15-year-old girl with familial adenomatous polyposis. AB - A 15-year-old girl with adenomatous polyposis coli gene (APC) mutation and brain tumor-polyposis syndrome developed an unusual succession of cervicocephalic tumors (medulloblastoma, meningeal low-grade myxoid tumor, and papillary thyroid carcinoma), at the age of 5, 9, and 15 years, respectively. We discuss the genetic profile of the thyroid tumor in which a large somatic deletion of APC gene was found and the physiopathology of thyroid carcinoma in patients with germline APC mutation. We also point out the uncommon phenotype in this young girl with early multiple neoplasias and the difficulties of management of such familial adenomatous polyposis patients with occurrence of extracolonic cancers that require the use of potential trigger agents as radiotherapy or chemotherapy. PMID- 19564753 TI - Serous cystadenocarcinoma of the ovary in an 11-year-old girl with extensive dissemination and chylothorax: a case report and a brief review of literature. AB - Ovarian neoplasms account for only 1% of all tumors in girls below the age of 17 years, and among these, malignant epithelial ovarian tumors are even more uncommon. We would like to report a case of ovarian epithelial malignancy presenting at 11 years of age with extensive peritoneal dissemination. The child ultimately succumbed to the illness and an autopsy was performed. PMID- 19564754 TI - The effect of anterior spinal fusion on spinal canal development in an immature porcine model. AB - STUDY DESIGN: Experimental study. OBJECTIVE: To investigate whether anterior spine fusion in the immature porcine spine has an adverse effect on the development of spinal canal. SUMMARY OF BACKGROUND DATA: Neurocentral cartilage (NCC) is located in the posterior vertebral body and responsible for the development of posterior aspect of the spinal canal. Injury to the NCC interferes with the development of the spinal canal. METHODS: Twelve 8-week-old domestic pigs were used to develop an anterior fusion model. A standard procedure as L3 L4, L4-L5 discectomy, and L3-L5 anterior instrumented spine fusion was performed. To evaluate the development of the spinal canal, all subjects had computed tomography scans before the procedure and at the final follow-up. The spinal canal area was measured at the control level (CL) (L2), arthrodesis level (AL) (L4), superior (L3), and inferior (L5) instrumented level (SIL and IIL). Percent change in spinal canal area from before surgery to final follow-up was also calculated. RESULTS.: Eleven subjects were available for the study. All subjects developed local kyphosis over the fused segments. The average area of L2 (CL) was 0.56 +/- 0.06 cm before surgery. The average areas of the L3 (SIL), L4 (AL), and L5 (IIL) were 0.62, 0.70, and 0.77 cm, respectively. At the final follow-up the average area of L2 was 1.20 cm. The average areas of the SIL, AL, and IIL were 1.16, 1.19, and 1.33 cm, respectively. The percent increase in spinal canal area at the CL was 116.6% whereas it was 85.8%, 71.0%, and 71.2% at SIL, AL, and IIL, respectively. CONCLUSION: Anterior spinal arthrodesis in the immature porcine spine results in iatrogenic retardation on spinal canal growth. This effect is most likely related to the tethering effect of the interbody fusion over the NCC. Although, it is difficult to directly extrapolate these findings to clinical practice, the spine surgeons operating on pediatric patients should be aware of this possibility. PMID- 19564755 TI - A novel biomechanical device improves gait pattern in patient with chronic nonspecific low back pain. AB - STUDY DESIGN: A retrospective study on patients with chronic nonspecific low back pain (NSLBP). OBJECTIVE: To describe the gait stride characteristics of patients with chronic NSLBP, and to examine the effect of a novel biomechanical device on the gait stride characteristics of these patients. SUMMARY OF BACKGROUND DATA: Patient with NSLBP alters their gait patterns. This is considered a protective mechanism as patients try to avoid extensive hip and spine ranges of motion and minimize forces and moments acting on the body. In addition, there are changes in the neuromuscular control system in patients with LBP that could possibly be attributed to the effects of pain on motor control. METHODS: Nineteen patients underwent a gait test, using an electronic walkway, at baseline and after 12 weeks of treatment. Spatiotemporal parameters were used to identify changes in gait pattern. A novel biomechanical device comprised of 4 modular elements attached to foot-worn platforms was used in the study. The modules are 2 convex shaped biomechanical elements attached to each foot, one is located under the hindfoot region and the other is located under the forefoot region. The device was individually calibrated to each patient. The patients were instructed to walk with the calibrated biomechanical device on a daily basis for a period of 12 weeks. RESULTS: Significant differences were found at baseline and after 12 weeks in normalized velocity (P = 0.03), cadence (P < 0.01), left normalized step length (P = 0.02), right normalized step length (P = 0.02), right swing (P < 0.01), right stance (P < 0.01), left single limb support (P = 0.01), left double limb support (P = 0.02), and right double limb support (P = 0.02). CONCLUSION: Patients with NSLBP treated with the novel biomechanical device for 3 months increased walking speed through longer step length and eliminated asymmetrical differences. PMID- 19564756 TI - Hematogenous spinal infection in central Greece. AB - STUDY DESIGN: We retrospectively analyzed spinal infection (SpI), in a teaching Hospital, in Central Greece. OBJECTIVE: To study presentation, etiology, and outcome of SpI in Central Greece. SUMMARY OF BACKGROUND DATA: SpI most frequently involves the intervertebral disc and adjacent vertebral bodies and can cause neurologic impairment. METHODS: Thirty three patients (23 men; age [mean +/- standard deviation], 60.6 +/- 11.3 years; disease duration, 44.5 [+/-54.7] days) hospitalized with SpI between January 2000 and December 2007 were included in the study. All patients had magnetic resonance imaging of the spine. RESULTS: Nineteen patients had pyogenic SpI (57.6%) and 14 patients had granulomatous SpI, 11 due to Brucella spp (34.4%), 3 due to Mycobacterium tuberculosis (9.4%). Staphylococcus aureus was the most frequent cause of pyogenic SpI, and spondylodiscitis (SpD) was the most frequent localization. Epidural entension was found in 8 of 17 pyogenic SpD and in 2 of 11 brucellar SpD patients. Subdural extension was detected in 3 patients with pyogenic SpD. Blood cultures were positive in 17 of 19 patients with pyogenic SpI. Two patients had concomitant endocarditis (staphylococcal 1, enterococcal 1). The most common associated disease was diabetes mellitus. All but 2 patients received medical treatment alone. Two patients died of uncontrollable sepsis. CONCLUSION: Back pain in presence of fever, constitutional symptoms, and/or high inflammation markers should alert physicians for spinal infection. In endemic areas, Brucella is a frequent cause of SpI. PMID- 19564757 TI - Can c7 plumbline and gravity line predict health related quality of life in adult scoliosis? AB - STUDY DESIGN: This study prospectively evaluated the health related quality of life (HRQOL) of 73 adults presenting with scoliosis at a single institution, as related to their spinal (C7 plumbline) and global (gravity line) balance. OBJECTIVE: To assess the influence of sagittal and coronal balance on HRQOL in adult scoliosis. SUMMARY OF BACKGROUND DATA: Many surgeons believe that achieving adequate spinal balance is important in the management of adult spinal deformity, but the evidence supporting this concept remains limited. A previous study has found weak correlations between sagittal spinal balance and HRQOL in adult spinal deformity, but this finding has never been confirmed independently. In addition, although the use of the gravity line is gaining interest in the evaluation of global balance, it remains unknown if this parameter is associated with HRQOL. METHODS: During a 1-year period, 73 consecutive new patients presenting with unoperated adult scoliosis and requiring full spine standing radiographs were evaluated using a force plate in order to simultaneously assess the gravity line. All patients also completed the Oswestry Disability Index (ODI) questionnaire to assess the HRQOL. Spinal balance was evaluated from the C7 plumbline and global balance from the gravity line, respectively. C7 plumbline and gravity line were both assessed with respect to the posterosuperior corner of the S1 vertebral body and central sacral vertebral line in the sagittal and coronal plane, respectively. C7 plumbline and gravity line, as well as their relative position, were correlated with the ODI, using Spearman coefficients. RESULTS: Sagittal spinal (C7 plumbline) and global (gravity line) balance, as well as their relative position were significantly related to the ODI. A poor ODI (>34) was associated with a sagittal C7 plumbline greater than 6 cm, a sagittal gravity line greater than 6 cm, and a C7 plumbline in front of the gravity line. Correlations between coronal balance and the ODI were not statistically significant. CONCLUSION: Sagittal spinal and global balance was strongly related to the ODI in adults with scoliosis. The observed correlation coefficients were higher than those reported in the only previous study suggesting the detrimental association of positive sagittal balance on ODI in adult spinal deformity. Coronal spinal and global balance did not influence the ODI in the current study cohort. Thisstudy underlines the relevance of C7 plumbline and gravity line in the evaluation of spinal and global balance, and lends further support to the philosophy of achieving adequate sagittal balance in the management of adult spinal deformity, especially in patients older than 50 years old with degenerative scoliosis. PMID- 19564758 TI - Surgical approach for synovial cyst of the atlantoaxial joint: a case report and review of the literature. AB - STUDY DESIGN: Case report and review of the literature. OBJECTIVE: We describe the first case of a synovial cyst arising from pseudarthrosis of a previous dens fracture. The literature is reviewed and etiological, diagnostic, and therapeutic options of atlantoaxial cysts are discussed. SUMMARY OF BACKGROUND DATA: Symptomatic synovial cysts of the atlantoaxial joint are rare. To the authors' knowledge only 24 cases have been reported.A 60-year-old patient presented with bilateral hand numbness, quadrihyperreflexia, and gait deterioration. Magnetic resonance imaging of the cervical spine disclosed a cystic mass located at the transverse ligament of dens axis causing bulbomedullary compression. METHODS: Surgery was performed via transoral image guided approach. The ventral atlas arch, dens, transverse ligament, tectorial membrane, and the compressing cyst were removed, followed by a C0-C3 fusion. RESULTS: Two months postsurgery the patient recovered completely from the cervical myelopathy with transient remnant dysparesthesia of the finger tips. CONCLUSION: Magnetic resonance imaging findings are not specific enough to establish a preoperative diagnosis. Radical resection via image-guided transoral route followed by posterior fusion allows complete resection of the cystic lesion and results in excellent long-term decompression. PMID- 19564759 TI - Multiple spinous process fractures of the thoracic vertebrae (Clay-Shoveler's Fracture) in a beginning Golfer: a case report. AB - STUDY DESIGN: A case report. OBJECTIVE: The authors report a unique experience with a 40-year-old male beginning golfer who presented with multiple isolated spinous process fractures of the upper thoracic spine. SUMMARY OF BACKGROUND DATA: Stress fractures are caused by repetitive loading and overuse, and they account for 10% of all sports-related injuries. Fractures of isolated spinous processes of the cervical and thoracic vertebrae are referred to as Clay Shoveler's fractures, and there is little clinical information on these injuries. METHODS: A 40-year-old man was admitted to our clinic with a 2-week history of posterior neck pain that radiated bilaterally into the shoulder area. Radiograph findings were normal. However, with a suspicion of fracture due to severe tenderness in the upper thoracic area, we performed three-dimensional cervicothoracic spine computed tomography and magnetic resonance imaging, which revealed multiple spinous process fractures at upper thoracic vertebrae. RESULTS: Conservative treatment was administered, and immobilization was maintained for more than 4 weeks. He has been symptom-free for more than 6 months. CONCLUSION: Although stress fractures of the spinous process are very rare and can be overlooked in clinical settings, they should be considered in the differential diagnosis of severe dorsal neck pain in golfers, especially beginners. PMID- 19564760 TI - Lumbar metastasis of choriocarcinoma. AB - STUDY DESIGN: A case of lumbar metastasis of a choriocarcinoma is presented. OBJECTIVE: To present and review a rare case of metastatic choriocarcinoma in the lumbar spine. SUMMARY OF BACKGROUND DATA: Choriocarcinoma is a highly anaplastic malignancy derived from trophoblastic cells characterized by the secretion of human chorionic gonadotropin (hCG) and early hematogenous metastasis. However, metastatic choriocarcinoma in the spine is extremely rare. Although 2 cases of metastasis in lumbar and/or sacral vertebra have been reported, the efficacy of surgical treatment for the spinal metastasis of choriocarcinoma is not yet known. METHODS: The clinical course, radiologic features, pathology, and outcome of the treatment of metastatic choriocarcinoma of the lumbar spine is reported. RESULTS: A 38-year-old female patient with abnormal uterine bleeding 6 weeks after a normal-term delivery showed high serum levels of hCG. A whole body image analysis revealed a lesion in the L2 vertebra. After computed tomography-guided needle biopsy, a clinical and pathologic diagnosis of lumbar metastasis of choriocarcinoma was made. Surgical resection of the localized L2 vertebra lesion was performed by total en bloc spondylectomy after a poor response to initial chemotherapy with methotrexate. Postsurgically, the serum level of hCG explosively increased and local recurrences around the original L2 vertebra and epidural metastasis abruptly developed. Lung metastases also occurred concurrently and progressed and the patient eventually died to the disease. CONCLUSION: We have reported a rare case of lumbar metastasis of choriocarcinoma after a normal-term pregnancy. This is the first report of lumbar metastasis of choriocarcinoma treated by spinal surgery. Because surgical resection of a lumbar metastasis of choriocarcinoma involves a substantial risk of profuse hemorrhage, local recurrence and the spread of metastasis, multiagent chemotherapy in combination with radiotherapy should be preformed before surgical resection. PMID- 19564761 TI - Effectiveness of the physical therapy Godelive Denys-Struyf method for nonspecific low back pain: primary care randomized control trial. AB - STUDY DESIGN: A simple blind, random controlled clinical trial. OBJECTIVE: To assess the effectiveness of physiotherapy treatment based on the muscular and articular chains Godelive Denys-Struyf (GDS) method for nonspecific low back pain (LBP) in primary care. SUMMARY OF BACKGROUND DATA: Despite a systematic review by the European COST ACTION B13 "Low back pain: guidelines for its management," there are still many unresolved questions regarding the effectiveness of the different physical therapy treatments used for LBP. SETTING: 21 physicians and physiotherapists in 7 Primary Care Centers and 6 researches in the Complutense University of Madrid (Spain). PARTICIPANTS: 137 patients diagnosed with nonspecific LBP. METHODS: The control group underwent 15 sessions of conventional physiotherapy in Primary Care Centers, and the experimental group received 15 GDS treatment sessions. Pain was evaluated by Visual Analogical Scale (VAS), functional disability by Oswestry questionnaire, and quality of life by the physical and mental components of SF-36 questionnaire. Outcome measures were assessed before treatment (A1), at the end of treatment (A2), and at 3 months (A3), and 6 months (A4) of follow-up. RESULTS: Repeated measures analysis of variance revealed that at the end of treatment and 3 months later, subjects in both groups showed less pain, reduced functional disability, and an improved quality of life, though improvements were greater in the GDS group.Six months after treatment, patients in the GDS group continued to show reduced pain (VAS(A4 A1) = -3.54, 95% CI: -4.18 to -2.90) while VAS scores in the control group returned to initial values (VAS(A4-A1) = 0.15, 95% CI: -0.36 to 0.67). CONCLUSION: Treatment of nonspecific LBP using the GDS method provides greater improvements in the midterm (6 months) in terms of the pain, functional ability, and quality of life perceived by patients than the conventional treatment based administered in primary care. PMID- 19564762 TI - Prevalence of sacral spina bifida occulta and its relationship to age, sex, race, and the sacral table angle: an anatomic, osteologic study of three thousand one hundred specimens. AB - STUDY DESIGN: An anatomic, osteologic study of spina bifida occulta (SBO). OBJECTIVE: To determine the prevalence and patterns of SBO in a large population and examine its relationship to age, sex, and race; then to evaluate SBOs relationship to the sacral table angle (STA) when compared with an age-matched control group. SUMMARY OF BACKGROUND DATA: SBO has a reported prevalence of 1.2% to 50% and has been implicated in various pathologic problems. SBO is often associated with spondylolysis or spondylolithesis. The STA has been implicated as an etiologic or predictive factor in the presence of pars defects. METHODS: Three thousand one hundred osteologic specimens were evaluated for the presence of SBO. SBO was graded on a scale from 0 to III. Information on the age, sex, race, and STA of each specimen was recorded and measured, respectively. Prevalence and patterns of SBO were enumerated. The STAs of an age-matched control group of 355 specimens were examined. The SBO group and control groups were compared in regards to STA, controlling for age, sex, and race. RESULTS: Overall, 355 specimens displayed SBO, for an overall prevalence of 12.4%. Of the SBO specimens, 68.7% were white, 88.2% were men, 53% were grade I, 37% II, and 10% III. All 3 grades of SBO were more common in men than women (88.2% vs. 11.8%) and more prevalent in whites than blacks (68.7% vs. 31.3%) (P = 0.01). SBO decreased in prevalence with increasing age. The average STA in SBO specimens was 95.9 degrees . This differed from an age-matched control group, 92.1 degrees (P < 0.0001). Every 1 degrees increase in STA resulted in a 6% increased likelihood of SBO. In SBO specimens, the STA decreased with increasing age, contrary to age matched controls. CONCLUSION: SBO has an overall prevalence of 12.4% in a large, diverse population. SBO is more common in men and whites and decreases in prevalence with increasing age. The STA is greater in SBO when compared with controls and an increased STA predicts SBO. In SBO, the STA decreases with increasing age. PMID- 19564763 TI - Up-regulation in receptor for advanced glycation end-products in inflammatory circumstances in bovine coccygeal intervertebral disc specimens in vitro. AB - STUDY DESIGN: This study investigated whether or not the receptor for advanced glycation end-products (RAGE) was up-regulated in inflammatory circumstances and consequently associated with aggrecan content in nucleus pulposus in vitro. OBJECTIVE: To investigate the activation of AGEs-RAGE complex by the irritation of IL-1beta in bovine intervertebral disc (IVD). SUMMARY OF BACKGROUND DATA: Although we have demonstrated that the accumulation of AGEs contributed to disc degeneration in human, it may be that acceleration in the AGEs-RAGE complex might be more important, mediated by expression levels of RAGE that increase in inflammatory mediators including IL-1beta in some tissues. Therefore, we investigated, in this study, the correlation if any between IL-1beta and AGEs RAGE complex in bovine IVD. METHODS: Samples of bovine coccygeal IVDs were harvested (n = 6). The presence of AGEs and RAGE were identified by immunohistochemistry. Real-time polymerase chain reaction (PCR) was used to quantify the messenger RNA levels of aggrecan after 6 days' stimulation of AGEs. Real-time PCR and immunofluorescein cytochemistry were performed to analyze the expression of RAGE after 2 days' stimulation of IL-1beta. The aggrecan expressions were evaluated by real-time PCR after 2 days' stimulation of combination of AGEs and/or IL-1beta. RESULTS: Immunohistochemical analysis revealed that AGEs and RAGE were localized within the bovine IVDs. AGEs significantly decreased the aggrecan expression in bovine IVD as in human IVD. The RAGE expression was significantly increased by 2 days' stimulation of IL 1beta. The aggrecan expression was decreased by stimulated AGEs and IL-1beta together, although not decreased by stimulated AGEs or IL-1beta separately. CONCLUSION: This is the first report to show the correlation between IL-1beta and AGEs-RAGE complex in IVD. Our results suggested that the increased RAGE expression in inflammatory circumstances and interaction with AGEs are risk factors in decreasing of aggrecan content in nucleus pulposus. PMID- 19564764 TI - In vitro proliferation and osteoblastic phenotype expression of cells derived from human vertebral lamina and iliac crest. AB - STUDY DESIGN: Osteoblastic cells derived from vertebral lamina and iliac crest were isolated and cultured under the same conditions (osteogenic medium, pH, temperature, and CO2 levels). OBJECTIVE: To compare proliferation and expression of osteoblastic phenotype of cells derived from vertebral lamina and iliac grafting. SUMMARY OF BACKGROUND DATA: Many factors play a role in the success of bone graft in spinal fusion including osteoblastic cell population. Two common sources of graft are vertebral lamina and iliac crest, however, differences in proliferation and osteoblastic phenotype expression between cells from these sites have not been investigated. METHODS: Cells obtained from cancellous bone of both vertebral lamina and iliac crest were cultured and proliferation was evaluated by direct cell counting and viability detected by Trypan blue. Alkaline phosphatase (ALP) activity was evaluated by thymolphthalein release from thymolphthalein monophosphate and matrix mineralization by staining with alizarin red S. Gene expression of ALP, osteocalcin, runt-related transcription factor 2, Msh homeobox 2, bone morphogenetic protein 7, intercellular adhesion molecule 1 precursor, osteoprotegerin, and receptor activator of NF-kB ligand was analyzed by real-time PCR. All comparisons were donor-matched. RESULTS: Proliferation was greater at days 7 and 10 in cells from vertebral lamina compared with ones from iliac crest without difference in cell viability. ALP activity was higher in cells from vertebral lamina compared with cells from iliac crest at days 7 and 10. At 21 days, mineralized matrix was higher in cells derived from vertebral lamina than from iliac crest. At day 7, gene expression of ALP, osteocalcin, runt related transcription factor 2, Msh homeobox 2, bone morphogenetic protein 7, intercellular adhesion molecule 1 precursor, receptor activator of NF-kB ligand, and osteoprotegerin was higher in cells derived from vertebral lamina compared with iliac crest. CONCLUSION: Cell proliferation and osteoblastic phenotype development in cells derived from cancellous bone were more exuberant in cultures of vertebral lamina than of iliac crest. PMID- 19564765 TI - Stress analysis of the interface between cervical vertebrae end plates and the Bryan, Prestige LP, and ProDisc-C cervical disc prostheses: an in vivo image based finite element study. AB - STUDY DESIGN: Segmental motion and bone-implant interface stresses were analyzed at C5-C6 levels with Bryan, Prestige LP, and ProDisc-C cervical disc prostheses using an image-based finite element modeling technique. OBJECTIVE: To predict stress patterns at the interface between prosthesis and lower vertebral end plate to better understand the underlying mechanisms of subsidence and how the load transfer pattern of each disc design affects segmental motion. SUMMARY OF BACKGROUND DATA: Subsidence is one of the most commonly reported device-related complications in intervertebral disc arthroplasty. Although clinical outcomes have been reported regarding many types of cervical prostheses, few reports have analyzed the effects of stress from cervical artificial discs to the vertebral end plate. METHODS: Three-dimensional voxel finite elements were built for C5-C6 spine unit based on computed tomography images acquired from a patient with indication for cervical disc arthroplasty. Models of facet joints and uncovertebral joints were added and artificial disc designs were placed in the intervertebral disc space. Static analyses were conducted under normal physiologic loads in flexion, extension, and lateral bending with precompression. RESULTS: Bryan disc recovered highest range of motion (4.75 degrees ) due to the high elastic nucleus, and therefore imposed the lowest stresses superior to C6. The ProDisc-C and Prestige LP discs caused high stress concentrations around their central fins or teeth, and may initiate bone absorption. Analysis of Prestige LP disc may indicate possible subsidence posteriorly caused by the rear positioned metal-to-metal joint. CONCLUSION: Rigidity of the cores ("nuclei") in Prestige LP and ProDisc-C prostheses guarantee initial maintenance of disc height, but high contact stress takes place at the bone-end plate interface if they are improperly placed or undersized. Anchorage designs add an additional factor that may increase propensity of subsidence, indicated by the high contact stress occurring at the end plate flanges of Prestige LP, and at midline keel fixation on the end plate of ProDisc-C. Although Bryan disc differs in these 2 concerns, it also creates much larger displacement during motion with more variation in disc height that may theoretically increase the load sharing of facet and/or uncovertebral joints compared to more rigid artificial discs. PMID- 19564766 TI - Dynamic plates in anterior cervical fusion surgery: graft settling and cervical alignment. AB - STUDY DESIGN: Retrospective cohort. OBJECTIVE: To evaluate the effect of graft subsidence associated with the use of dynamic cervical plates on cervical alignment and clinical outcome of patients undergoing anterior cervical decompression and fusion. SUMMARY AND BACKGROUND DATA: Dynamic cervical plates allow graft loading which may enhance fusion. However, subsidence of the bone graft may result in changes in cervical alignment. The extent of settling of the graft and its impact on cervical alignment has not been determined. Also whether such change in alignment has an impact on clinical outcome is not known. METHODS: Fifty-five patients with spondylitic myelopathy or radiculopathy underwent anterior cervical decompression and fusion using a dynamic plate for stabilization (ABC, Braun/Aesculap). The surgical indications included myelopathy (n = 21), radiculopathy (n = 30), or myeloradiculopathy (n = 4). Regional and local cervical angles, graft subsidence, and fusion rates were assessed before surgery, immediately after surgery and at 6 months after surgery. Visual analogue scores for neck and radicular pain, Neck Disability Indexes, and Nurick grades for myelopathy were recorded. Patients were observed up for at least 6 months with a median of 14 months (range: 6-48 months). RESULTS: Radicular pain improved by a median of 5 points (P < 0.0001), whereas Neck pain improved by a median score of 3 (P < 0.0001). A median improvement in Neck Disability Index score of 18 (P < 0.0001) was observed. Nurick grades improved in 68% of patients with myelopathy. Fusion was achieved in 96% of cases. The mean graft subsidence at 6 months was 1.7 mm (range: 0-6 mm). A change of local cervical angle toward lordosis of 5.8 degrees (P < 0.0001) was obtained as a result of surgery. However, there was a loss of 2.7 degrees of lordosis in the first 6 months following surgery (P = 0.001). The extent of subsidence correlated with the local loss of lordosis (P = 0.0003). There was no change in the regional cervical angle in the 6 months following surgery. There was no significant association between clinical outcome and changes in cervical angles. CONCLUSION: Dynamic anterior cervical plating after anterior decompression and grafting provides comparable fusion rates to that reported following the use of rigid cervical plating. Immediate changes in cervical alignment towards lordosis are partially lost on follow-up. The loss of lordosis is related to the amount of graft settling. PMID- 19564767 TI - Centering of cervical disc replacements: usefulness of intraoperative anteroposterior fluoroscopic guidance to center cervical disc replacements: study on 20 discocerv (scient'x prosthesis). AB - STUDY DESIGN: This is a prospective randomized computed tomographic scan study on the centering of cervical disc prosthesis (Discocerv; Scient'X) with and without anteroposterior (AP) fluoroscopic guidance. OBJECTIVE: Analyze interest of AP fluoroscopic guidance for coronal positioning in cervical disc replacements. SUMMARY OF BACKGROUND DATA: This series consisted of 20 patients. One group of 10 patients was operated using only lateral fluoroscopic guidance (L guidance) and the other group of 10 patients was operated using both lateral and AP fluoroscopic guidance (AP + L guidance). Total disc replacements positioning is analyzed in the 2 groups. METHODS: All patients had a computed tomographic scan 24 hours after surgery. Specific reconstructions were obtained from the native slices. Three planes P1, P2, and P3 are defined to quantify centering of the prosthesis in axial sagittal and coronal planes. RESULTS.: In the coronal plane P1, there is no difference in lateralization between the L guidance (absolute value of average M = 0.93 mm; SD = 0.59 mm) and AP + L guidance groups (M = 1.28 mm; SD = 0.75 mm). In the axial plane, there is no difference in lateralization between the L guidance and AP + L guidance groups. In the L guidance group, average was 1.96 degrees (SD = 1.43 degrees ) and 3.18 degrees (SD = 2.94 degrees ) in AP + L guidance. There is no significative difference between 2 groups in coronal (P = 0.26) and axial plane (P = 0.19). CONCLUSION: Unci are reliable landmarks for coronal centering of total disc replacements. AP fluoroscopic guidance does not improve this positioning. PMID- 19564768 TI - A correlation of radiographic and functional measurements in adult degenerative scoliosis. AB - STUDY DESIGN: Retrospective functional and radiographic analysis of symptomatic patients with de novo degenerative lumbar and thoracolumbar scoliosis. OBJECTIVE: To evaluate the radiographic parameters of symptomatic patients presenting with de novo degenerative adult scoliosis and correlate them with functional scores. SUMMARY OF BACKGROUND DATA: Previous studies have been inconclusive as to the correlation of radiographic parameters and clinical symptomatology. METHODS: Radiographic analysis of 58 consecutive symptomatic patients with de novo degenerative lumbar and thoracolumbar scoliosis was performed using posteroanterior and lateral 36-inch standing radiographs. Measurements included curve type, curve location, curve magnitude, coronal alignment, sagittal alignment, and anteroposterior and lateral intervertebral olisthesis. Clinical functional data were measured with Oswestry Disability Index, Roland-Morris Disability Questionnaire, and RAND 36-item Health Survey questionnaire. Correlation between clinical data and radiographic data were then calculated. RESULTS: Sagittal balance did not show significant correlation with functional results. However, coronal imbalance (more than 5 cm from midsacrum) affected physical function (P = 0.028) and outcomes (P > 0.05). Also, moderate to severe lateral olisthesis (equal or more than 6 mm) demonstrated higher bodily pain then mild lateral olisthesis (P = 0.005). Good lumbar lordosis correlated positively with health assessment as reflected in SF-36 score (P = 0.039, r = 0.291). CONCLUSION: Reduced lumbar lordosis and increased lumbosacral scoliosis can affect the general health status of older patients with de novo degenerative scoliosis. Lateral olisthesis, mainly, and anteroposterior olisthesis are important elements of rotatory subluxation in the lumbar curves, which are important radiographic parameters, predicting symptomatology and health status of patients with de novo degenerative scoliosis. PMID- 19564769 TI - The flip test: a reappraisal. AB - STUDY DESIGN: Diagnostic study. OBJECTIVE: To assess the construct validity of the flip test as a sign of nerve root tension. SUMMARY OF BACKGROUND DATA: The flip test is commonly performed in patients with sciatica to confirm or otherwise nerve root tension evidenced by a restricted supine straight leg raise (SLR). Passive extension of the knee with the patient in the erect position and the hip flexed is reported to cause a sudden falling or flipping back of the trunk. Although widely known there are no research articles validating this test. METHODS: Sixty-seven patients with sciatica and magnetic resonance imaging scans confirming disc protrusion and nerve root compression underwent the flip test. The response of the patient was recorded, compared with the supine SLR, and subjected to statistical analysis. RESULTS: Thirty-three percent of patients felt no pain, 39% felt pain on full extension of the knee, and 28% resisted full extension of the knee due to pain. Only one-third of patients demonstrated a "flip." The response was related to the supine SLR, such that patients with high SLRs tended not to show a painful reaction whereas all patients with a supine SLR below 45 degrees showed a painful response. CONCLUSION: We consider that the flip test has been substantially misinterpreted in the past. However, it remains a useful check of nerve root tension but only for patients with supine SLRs below 45 degrees . The most reliable response was not a flip but the demonstration of pain on extension of the knee. We would recommend the term "sitting SLR test," as a more accurate and less misleading name. PMID- 19564770 TI - Orthopaedists' and family practitioners' knowledge of simple low back pain management. AB - STUDY DESIGN: Comparative knowledge survey. OBJECTIVE: This study compared the knowledge of orthopaedic surgeons and family practitioners in managing simple low back pain (LBP) with reference to currently published guidelines. SUMMARY OF BACKGROUND DATA: LBP is the most prevalent of musculoskeletal conditions. It affects nearly everyone at some point in time and about 4% to 33% of the population at any given point. Treatment guidelines for LBP should be based on evidence-based medicine and updated to improve patient management and outcome. Studies in various fields have assessed the impact of publishing guidelines on patient management, but little is known about the physicians' knowledge of the guidelines. METHODS: Orthopedic surgeons and family practitioners participating in their annual professional meetings were requested to answer a questionnaire regarding the management of simple low back pain. Answers were scored based on the national guidelines for management of low back pain. RESULTS: One hundred forty family practitioners and 253 orthopaedists responded to the questionnaire. The mean family practitioners' score (69.7) was significantly higher than the orthopaedists' score (44.3) (P < 0.0001). No relation was found between the results and physician demographic factors, including seniority. Most orthopaedists incorrectly responded that they would send their patients for radiologic evaluations. They would also preferentially prescribe cyclo-oxygenase 2-specific nonsteroidal anti-inflammatory drugs, despite the guidelines recommendations to use paracetamol or nonspecific nonsteroidal anti-inflammatory drugs. Significantly less importance was attributed to patient encouragement and reassurance by the orthopaedists as compared with family physicians. CONCLUSION: Both orthopaedic surgeons' and family physicians' knowledge of treating LBP is deficient. Orthopedic surgeons are less aware of current treatment than family practitioners. Although the importance of publishing guidelines and keeping them up-to-date and relevant for different disciplines in different countries cannot be overstressed, disseminating the knowledge to clinicians is also very important to ensure good practice. PMID- 19564771 TI - Influence of time, activities, and memory on the assessment of chronic low back pain intensity. AB - STUDY DESIGN: A prospective study. OBJECTIVE: To identify factors potentially influencing patient self-evaluation of chronic low back pain (CLBP) and their interactions. SUMMARY OF BACKGROUND DATA: The impact of the time, context, and/or recall on the self-assessment of chronic pain intensity remains poorly understood. METHODS: A prospective study investigated patients' CLBP intensity over 29 days. Patients were evaluated on days 1 and 29 by the investigator with a pain intensity visual analog scale (0-100), and interviewed by phone on days 8, 15, and 22 to 28 to evaluate pain intensity with a numerical rating scale (0-10), either thrice daily (time-associated group), or during rest/activities (activity associated group). Current and/or recalled pain intensities were also assessed. Means of recorded pain intensity and overall assessment of patient's pain intensity of the day were subjected to statistical analyses with analysis of variance and test of correlation. RESULTS: Among the 203 patients included, 194 (56.2% women; mean age: 47.5 +/- 12.1 year) were considered in the analysis (9 patients were not followed until day 24). Strong correlations were obtained between mean pain intensity (time-associated group = 44 patients) and overall assessment of pain intensity of the day. Strong correlations were also obtained between overall assessment of pain intensity of the day and intensity measured during each activity, but not between current pain intensity and remembered pain 24 of 48 hours earlier (activity-associated group = 150 patients). Recalled pain intensities for the last 7 or 28 days were strongly correlated with current pain intensity, but the correlations were markedly weaker after adjustment for day 29 pain intensity. CONCLUSION: For CLBP patients, overall assessment of pain intensity of the day at evening is accurate to assess pain on 1 day. Overall assessment of pain intensity of the day is very close with usual pain. Furthermore, 24 and 48 hours remembered pain intensity are not accurate measures. Recalls of pain on the 7 or 28 last days were very dependant of the pain intensity of the day of the assessment. These findings could contribute to improving pain intensity assessment in CLBP patients avoiding multiple assessments, especially during clinical trials. PMID- 19564772 TI - Discriminating healthy controls and two clinical subgroups of nonspecific chronic low back pain patients using trunk muscle activation and lumbosacral kinematics of postures and movements: a statistical classification model. AB - STUDY DESIGN: Statistical Classification Model for nonspecific chronic low back pain (NS-CLBP) patients and controls based on parameters of motor control. OBJECTIVE: Develop a Statistical Classification Model to discriminate between 2 subgroups of NS-CLBP (Flexion Pattern [FP] and Active Extension Pattern [AEP]) and a control group using biomechanical variables quantifying parameters of motor control. SUMMARY OF BACKGROUND DATA: It has been well documented that many CLBP patients have motor control impairments of their lumbar spine. O'Sullivan proposed a mechanism-based classification system for NS-CLBP with motor control impairments based on a comprehensive subjective and physical examination to establish the relationship between pain provocation and spinal motor control. For the FP and AEP s, 2 groups defined by O'Sullivan and under investigation is this study, the motor control impairment is considered to be the mechanism maintaining their CLBP. No previous studies have used a Statistical Model with measurements of motor control impairment to subclassify NS-CLBP patients. METHODS: Thirty three NS-CLBP patients (20 FP and 13 AEP) and 34 asymptomatic subjects had synchronized lumbosacral kinematics and trunk muscle activation recorded during commonly reported aggravating postures and movements. Biomechanical variables were quantified and a Statistical Classification Model was developed. RESULTS: The Statistical Model used 5 kinematic and 2 electromyography variables. The model correctly classified 96.4% of cases. CONCLUSION: Selected biomechanical variables were predictors for subgroup membership and were able to discriminate the 3 subgroups. This study adds further support toward the validation of the proposed classification system. PMID- 19564773 TI - Spinal myxopapillary ependymoma: neurological deterioration in patients treated with surgery. AB - STUDY DESIGN: A retrospective multicenter study of series of 20 patients with myxopapillary ependymomas, which underwent surgery. OBJECTIVE.: To evaluate the postoperative outcomes of patients surgically treated for spinal myxopapillary ependymoma. Neurologic deterioration after tumor resection is discussed in terms of the tumor encapsulation. SUMMARY OF BACKGROUND DATA.: Myxopapillary ependymomas occur most commonly in the cauda equina and/or conus medullaris region. Most series published in the literature have covered postoperative tumor recurrence and role of adjuvant radiotherapy. Few contradistinctive studies in postoperative neurological deficit were reviewed from the standpoints of MRI and histopathologic findings. METHODS: A total of 20 patients were recruited and charts, MRI, intraoperative findings, and pathologic findings were reviewed. The follow-up period ranged from 2 to 12 years (median, 72.9 months). The surgical procedures were defined as gross total removal, piecemeal total removal, and subtotal removal. Postoperative radiotherapy was given in 3 patients in whom removal had been subtotal. RESULTS: Gross total removal was achieved in 14 patients, piecemeal gross total and subtotal removal in 3 patients each. Neurologic deterioration after surgery was seen in 5 patients, all of which were unencapsulated tumors consisting of piecemeal gross total removal in 2 patients and subtotal removal in 3 patients. Recovery of postoperative bladder dysfunction remained unchanged in 2 patients. There were no tumor recurrence and progression of the remaining tumors. Unencapsulated tumors were more frequently seen in heterogeneously enhanced tumors on MRI than in homogenously enhanced tumors with significant difference. CONCLUSION: In the unencapsulated ependymomas, tumor separation and manipulation of the surrounding neural tissue caused neurologic injury. The heterogeneously enhanced ependymoma not only should be evaluated and treated meticulously, but also surgeons should not stick to total removal in infiltrated and adhering tumors as subtotally resected tumors with postoperative radiotherapy have not always recurred. PMID- 19564774 TI - Re: Myran R, Kvistad KA, Nygaard OP, et al. Magnetic resonance imaging assessment of the alar ligaments in whiplash injuries. Spine 2008;33:2012-6. PMID- 19564775 TI - Re: Kanayama M, Hashimoto T, Shigenobu K, et al. A minimum 10-year follow-up of posterior dynamic stabilization using Graf artificial ligament. Spine 2007;32: 1992-6. PMID- 19564776 TI - Re: Yang JY, Song HS, Lee M, et al. Adjacent level ossification development after anterior cervical fusion without plate fixation. Spine 2009;34:30-3. PMID- 19564778 TI - Re: Dhawan A, Klemme WR, Polly DW Jr. Thoracic pedicle screws: comparison of start points and trajectories. Spine 2008;33:2675-81. PMID- 19564780 TI - Re: Staal JB, de Bie R, de Vet HC, et al. Injection therapy for subacute and chronic low-back pain. Cochrane Database Syst Rev 2008:CD001824. PMID- 19564783 TI - Efficacy and safety of transarterial chemoembolization in recurrent hepatocellular carcinoma after curative surgical resection. AB - OBJECTIVES: To evaluate the efficacy and safety of transarterial chemoembolization (TACE) for recurrent hepatocellular carcinoma (HCC) after curative resection and to examine the factors associated with the prognosis in a single center. METHODS: Between January 1995 and July 2006, 169 patients who had undergone curative resection were diagnosed with recurrent HCC. Of these, 114 patients underwent TACE as first-line therapy and were followed until July 2007. RESULTS: At the time of resection, the mean tumor size was 4.8 + or - 2.9 cm, and 96 (84.2%) patients had a single tumor. Single nodular recurrence was observed in 53 (46.5%) patients. The mean size of the recurrent HCC was 2.1 + or - 1.2 cm. The disease-free survival after TACE was 46.0%, 16.7%, and 13.4% at 1, 2, and 3 years, respectively. The overall survival after TACE was 77.8%, 53.6%, and 31.6% at 1, 3, and 5 years, respectively. Cox regression analysis revealed that the Edmonson grade and time to recurrence (>6 months) independently affected the disease-free survival (both P < 0.05). The time to recurrence (>6 months) and tumor-node-metastasis stage were associated with overall survival (both P < 0.05). Only 1 lethal complication (biliary sepsis) occurred after TACE. CONCLUSIONS: TACE seems to be a safe, effective treatment for recurrent HCC after curative resection. PMID- 19564784 TI - A dosimetric analysis comparing treatment of low-risk prostate cancer with TomoTherapy versus static field intensity modulated radiation therapy. AB - OBJECTIVES: Static field intensity modulated radiation therapy (IMRT) has demonstrated dosimetric and clinical benefits over 3-dimensional conformal radiation therapy. TomoTherapy is a unique form of IMRT that may offer further improvements. METHODS: The study population consisted of 15 patients with low risk prostate cancer treated at Rush University with TomoTherapy (n = 7) or IMRT (n = 8). For each patient, both a TomoTherapy plan and an IMRT plan were generated using identical planning objectives. The planning target volume (PTV) was defined as the prostate and proximal seminal vesicles plus a margin. The prescription dose was 7740 cGy in 43 fractions. Radiation Therapy Oncology Group (RTOG) normal tissue guidelines were used as constraints, and the PTV coverage was made equivalent for the paired plans by equalizing the PTV V100. RTOG benchmark DVH values for the rectum and bladder and mean dose to the penile bulb were recorded. The volume of PTV receiving >= 105% of the prescription dose was measured. RESULTS: The mean DVH values for each of the RTOG constraints for rectum and bladder were significantly improved using TomoTherapy. The volume of the PTV that received at least 105% of the dose was higher with IMRT (11.7% vs. 0.2%, <0.001). The mean dose to the penile bulb was higher with TomoTherapy (40.4 Gy vs. 27.4 Gy, P = 0.005). CONCLUSIONS: TomoTherapy offers a more favorable dose distribution to the bladder and rectum, as well as improved target homogeneity in comparison with IMRT. PMID- 19564785 TI - Hormone receptor expression is associated with a unique pattern of metastatic spread and increased survival among HER2-overexpressing breast cancer patients. AB - OBJECTIVES: HER2/neu (HER2) overexpression occurs in approximately 20% of breast cancers and is associated with aggressive disease. Although a significant number of HER2-positive tumors also express hormone receptors (HR), the effects HR expression has on clinical characteristics, including response to trastuzumab among HER2-positive breast cancer, has not been elucidated yet. METHODS: A retrospective analysis of consecutive metastatic HER2-positive breast cancer patients was conducted in 2 medical centers. Associations between hormone receptors expression and clinical variables, and metastatic spread pattern and survival were studied. RESULTS: The study population included 137 metastatic HER2 positive breast cancer patients, 56 of them were HR-positive and 81 were HR negative. No significant differences between the 2 groups were found for demographic and clinical characteristics, including age, stage at diagnosis, tumor histology, and grade. Similar response rate to trastuzumab was observed in both study groups. Significantly, longer, median, disease-free, and overall survival was noted among the HR-positive patients. Patients in the HR-negative group had significantly more liver metastases, a trend for more brain metastases, and less bone metastases. There was a strong trend for more visceral metastases in the HR-negative group. CONCLUSIONS: Our results suggest an important role for HR expression in modulating metastases predilection and disease progression in HER2-positive breast cancer. PMID- 19564786 TI - Is Toxoplasma gondii a causal agent in migraine? AB - BACKGROUND: Many different tissues may be parasitized by Toxoplasma gondii, particularly, lung, heart, lymphoid organs, and the central nervous tissues. Tissue cysts of this parasite in the brain may spontaneously rupture, releasing parasites that cause antibody titers to rise. In immunocompetent subjects with acquired toxoplasmosis, the most frequent symptoms were lymphadenopathy and headache. In the neurogenic inflammation theory of the pathogenesis of migraine, the cause of initial release of ions and inflammatory agents has not been established. In this study, we aimed to investigate if T. gondii infection is a possible cause of neurogenic inflammation of migraines. METHODS: The anti-T. gondii antibody status of 104 patients with migraine were studied and compared with those of control groups, 50 healthy subjects and 50 subjects with headache due to rhinosinusitis, by using a micro-enzyme-linked immunosorbent assay technique. RESULTS: Forty-six (44.2%) patients with migraine, 13 (26.0%) healthy control subjects, and 12 (24%) control subjects with rhinosinusitis were positive for anti-T. gondii IgG antibody. The rate of positivity in the migraine patient group was statistically different from those of the control groups (P < 0.05). CONCLUSIONS: The results show the presence of chronic Toxoplasma infection in patients with migraine. Toxoplasma infection may contribute to neurogenic inflammation as the pathogenesis of migraine, as many studies in the literature have reported that Toxoplasma infection causes biochemical and immunologic changes. PMID- 19564787 TI - Delayed pericarditis and cardiac tamponade associated with active-fixation lead pacemaker in the presence of mitochondrial myopathy and Ockham's razor. AB - A 23-year-old male patient, with a diagnosed mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes, was referred for recurrent fainting. Long sinus pauses were documented, and an atrial pacemaker with an active-fixation lead was implanted. He was admitted again 4 months later because of chest pain and diffuse ST segment changes. On the basis of these, pericarditis was diagnosed, corticosteroid therapy and the adjunct of salicylates were started, which in few hours enabled the relief of symptoms and the reduction of ECG abnormalities. However, 24 h later, the patient suddenly experienced severe hypotension and tachycardia, and an emergency echocardiogram showed pericardial tamponade. The differential diagnoses with atrial free-wall perforation and Dressler-like syndrome were discussed, along with the difficulties in management. By a 'wait and see' strategy, the active-fixation atrial lead was eventually changed into a passive-fixation one, while continuing corticosteroids and salicylates. The patient quickly improved and is now, after 1 year, symptom free. For the explanation of any phenomenon, it is important that as few assumptions as possible are considered, eliminating those that make no difference in the observable predictions of the explanatory hypothesis or theory, according to the concept of Ockham's razor. PMID- 19564788 TI - The need for new oral anticoagulants in clinical practice: an introduction. PMID- 19564789 TI - Biography: Roy K. H. Wong, MD. PMID- 19564790 TI - Effect of HIV infection on the prevalence of colorectal adenomas during screening colonoscopy. PMID- 19564791 TI - Barrett's esophagus surveillance in Asians. PMID- 19564792 TI - The role of allergy evaluation in adults with eosinophilic esophagitis. AB - BACKGROUND: Allergy evaluation and skin prick testing for foods are recommended in all patients with eosinophilic esophagitis. However, the yield of allergy evaluation in adult eosinophilic esophagitis is not known. GOALS: To determine the yield of allergy evaluation in adults with eosinophilic esophagitis. STUDY: All adult patients in the Cleveland Clinic eosinophilic esophagitis registry between January 2006 and April 2008 were identified. Diagnosis was based on clinical presentation and endoscopy with biopsies showing > or = 15 eosinophils/hpf. From this group, all patients referred for allergy evaluation were identified. Allergy evaluation consisted of skin testing to foods in all patients. Selected patients underwent skin testing to inhalants based on the presence of concomitant allergic rhinitis and/or asthma. Immediate hypersensitivity skin testing was performed and scored by standard methodology. Patients were referred on an ad hoc basis by the gastroenterologist and not based on a priori identification of an allergic diathesis. RESULTS: Twenty-six out of 68 patients (38%) completed allergy evaluation. Thirteen out of 26 patients (50%) demonstrated a positive skin test to > or = 1 food allergen. Of the 15 patients who underwent skin testing for inhalants, 14 (93%) had a positive skin test to 1 or more inhalants. In total 21/26 patients (81%) had > or = 1 allergen identified, 16/26 (62%) had > or = 5 allergens identified, and 4/26 (15%) had > or = 10 allergens identified (range: 0 to 20 allergens identified). Peanut, egg white, soybean, cow's milk, and tree nuts were the most common food allergens identified. CONCLUSIONS: Allergy evaluation has a high yield in adult eosinophilic esophagitis as 81% of referred patients had one or more allergens identified and 50% had one or more skin tests positive to foods. Allergy evaluation should be considered in adult patients with eosinophilic esophagitis. PMID- 19564793 TI - Incidence of and predictive risk factors for intraductal papillary mucinous neoplasm of the pancreas with ordinary pancreatic cancer. PMID- 19564794 TI - Nebivolol induces parallel improvement of left ventricular filling pressure and coronary flow reserve in uncomplicated arterial hypertension. AB - PURPOSE: Our aim was to analyze the effects of 3-month antihypertensive therapy by nebivolol, a beta-blocking agent with nitric oxide-mediated vasodilatory properties, on coronary flow reserve (CFR) and left ventricular filling pressure (LVFP) in uncomplicated arterial hypertension. METHODS: Twenty newly diagnosed, never treated, uncomplicated hypertensive patients (14 male and six female patients, mean age = 49 years), I-II WHO grade, underwent single-blind nebivolol treatment. At baseline and at 3-month follow-up, patients underwent Doppler echocardiography including pulsed Tissue Doppler of septal mitral annulus: the ratio between transmitral E velocity and myocardial early diastolic velocity (E/Em ratio) was calculated as an index of LVFP degree. Transthoracic Doppler derived CFR (high-dose dipyridamole coronary diastolic peak flow velocity to resting coronary peak flow velocity ratio) of distal left anterior descending artery was also determined. RESULTS: After 3-month nebivolol therapy, rate pressure product decreased (P < 0.0001). No significant change of left ventricular mass index, relative wall thickness and midwall shortening was detected. Left ventricular end-diastolic diameter and stroke volume were both marginally increased. Nebivolol increased Em (P < 0.0001), reduced E/Em ratio (from 9.0 +/- 1.6 to 8.2 +/- 1.1, P < 0.0001) and enhanced CFR (from 2.07 +/- 0.2 to 2.20 +/- 0.2, P = 0.003), because of increased hyperemic coronary flow velocity (P < 0.001). CFR increase remained significant (P < 0.001) after normalizing resting and dipyridamole coronary velocities for the respective rate pressure product. The increase of normalized CFR induced by nebivolol was related with E/Em ratio decrease (r = -0.65, P < 0.002). CONCLUSION: Nebivolol improves LVFP as well as CFR in uncomplicated hypertension. The association between changes of CFR and those of LVFP indicates a possible common denominator between improvement of coronary microvascular function and myocardial stimulation of nitric oxide release induced by the drug. PMID- 19564795 TI - Bottom blood pressure or bottom cardiovascular risk? How far can cardiovascular risk be reduced? AB - BACKGROUND: Recent intervention trials have been conducted in patients at high cardiovascular risk, and their results have reopened the issue whether lowering blood pressure to normotensive values is of greater benefit than lowering blood pressure below 140/90 mmHg. These trials have made widespread use of concomitant therapies (lipid-lowering, antiplatelet and background antihypertensive agents). The question has been addressed whether in these trials a bottom level of cardiovascular risk (i.e. one that cannot be further reduced) rather than a bottom level of blood pressure (i.e. one below which risk cannot be further reduced) was achieved. METHODS: The 'residual risk', that is, the incidence of major cardiovascular events achieved in trials with antihypertensive agents, was calculated by reviewing endpoint data in all major trials after classifying them into four categories according to patients' baseline cardiovascular risk: low risk patients; elderly patients; diabetic patients; high-risk patients. RESULTS: Low rates of major cardiovascular events (below 3-6% in 5 years) were only achieved in trials enrolling low-risk patients. In elderly hypertensive patients, hypertensive patients with diabetes and particularly patients with previous cardiovascular disease quite rarely could incidence of major cardiovascular events be reduced below a bottom level of 12-14% in 5 years, and remained within the high-risk range (above the conventional threshold of 10% in 5 years) despite extensive use of concomitant therapies. CONCLUSION: In high-risk patients there is a 'ceiling effect' for treatment benefits. Delaying therapeutic correction of cardiovascular risk factors until a high level of risk is achieved blunts the full benefits of interventions. PMID- 19564796 TI - Are ADHD symptoms associated with delay aversion or choice impulsivity? A general population study. AB - OBJECTIVE: The term delay aversion has been used both to describe a behavioral tendency of greater preference for smaller-immediate over larger-delayed rewards (choice impulsivity) and to refer to a secondary explanatory construct put forward by delay aversion theory. In this study, we examined the association of attention-deficit/hyperactivity disorder (ADHD) symptoms with choice impulsivity and tested the specific hypothesis derived from delay aversion theory. METHOD: A total of 1,062 children aged 7.90 to 10.90 years (49% girls) made a fixed number of repeated choices between a smaller reward delivered immediately and a larger reward delivered after a delay (choice-delay task), under two conditions (including and excluding a postreward delay). We assessed the unique contribution of each ADHD symptom dimension to the prediction of choice impulsivity and delay aversion, controlling for age (or age and IQ). Sex effects were examined. RESULTS: : Inattention ratings uniquely predicted preference for smaller immediate rewards under both task conditions for both sexes. An index of delay aversion was associated with inattention only in boys; the effect size was small yet significant. Hyperactivity-impulsivity ratings were negatively associated with choice impulsivity in girls in the postreward delay condition, whereas no significant association with hyperactivity-impulsivity ratings was observed in boys. Categorical analyses using groups with high ADHD symptoms yielded similar results. CONCLUSIONS: This is the first study to report a unique association between inattention symptoms and behavioral measures of choice impulsivity and delay aversion. The findings indicate the importance of the primary constitutional processes that underlie choice impulsivity and their potential role in behavioral inattention. Understanding the behavioral and brain processes underlying choice impulsivity may lead to the improved targeting of behavioral and pharmacological interventions. PMID- 19564797 TI - Childhood risk factors for lifetime anorexia nervosa by age 30 years in a national birth cohort. AB - OBJECTIVE: To examine whether previously identified childhood risk factors for anorexia nervosa (AN) predict self-reported lifetime AN by age 30 years in a prospective birth cohort. METHOD: Using data from the 1970 British Cohort Study, at birth, 5, 10, and 30 years, we examined associations between suggested childhood risk factors and self-reported lifetime AN at 30 years, adjusted for sex and socioeconomic status. RESULTS: Anorexia nervosa was independently predicted by female sex (odds ratio [OR] 22.1), infant feeding problems (OR 2.6), maternal depressive symptoms (OR 1.8), and a history of undereating (OR 2.7). High self-esteem (OR 0.3) and higher maternal body mass index (OR 0.91) were protective. CONCLUSIONS: Of 22 suggested risk factors for AN, only four risk and two protective factors were confirmed in this prospective cohort study. We did not identify a role for perinatal factors, parenting, childhood body mass index, childhood emotional or sleep problems, academic ability, or exercise. PMID- 19564798 TI - Anticipating problem drinking risk from preschoolers' antisocial behavior: evidence for a common delinquency-related diathesis model. AB - OBJECTIVE: Early first drinking (EFD) experiences predict later alcohol problems. However, the longitudinal pathway from early childhood leading to EFD has not been well delineated. Based on documented links between drinking behaviors and chronic antisocial behaviors, this article tests a common diathesis model in which precursive patterns of aggression and delinquent behavior-from preschool onward-anticipate EFD. METHOD: Participants were 220 male children and their parents in a high risk for substance use disorder prospective study. Early first drinking was defined as having had a first drink by 12 to 14 years of age. Stacked structural equation models and configural frequency analyses were used to compare those with and without EFD on aggression and delinquent behavior from ages 3 to 5 years through 12 to 14 years. RESULTS: Delinquent behavior and aggression decreased normatively throughout childhood for those with and without EFD, although those with EFD showed precocious resurgences moving into early adolescence. Early first drinking was associated with delinquent behavior more than aggression. Early drinkers were more delinquent at most ages-with a direct effect of preschool predisposition on adolescent behavior only within the EFD group. Early first drinking was disproportionately likely among individuals with high levels of delinquent behavior at both 3 to 5 and 12 to 14 years of age but uncommon among individuals with low levels of delinquent behavior during those two age periods. CONCLUSIONS: Early first drinking and delinquent behavior share a common diathesis evident before school entry. Intervention and prevention programs targeting problem drinking risk should focus on dismantling this emergent primarily delinquency-related developmental trajectory. PMID- 19564799 TI - Risk factors for full- and partial-syndrome early adolescent eating disorders: a population-based pregnancy cohort study. AB - OBJECTIVE: To identify prospective predictors of eating disorders in a population based sample of 14-year-old boys and girls, using previously collected antenatal, biomedical, familial, demographic, and psychosocial data. METHOD: Participants (N = 1,597) were drawn from the Western Australian Pregnancy Cohort (Raine) Study. Data were collected during pregnancy, at birth, and when children were aged 1, 2, 5, 8, 10, and 14 years. An adapted version of the Eating Disorder Examination Questionnaire was used to assess eating disorder symptoms at age 14 years. Logistic regression was used to identify prospective predictors of eating disorder caseness, relative to general control and psychiatric control groups. RESULTS: At age 14 years, 6% of the sample met full or partial criteria for a DSM IV eating disorder. Being female and being perceived as overweight by one's parent were the strongest predictors of eating disorder caseness in the final multivariate models, relative to both control groups. Maternal body mass index, social problems, low social-related self-efficacy, and neurocognitive difficulties were also predictive of eating disorder caseness relative to the general control group only. CONCLUSIONS: The results suggest that parent's perceptions of their child's weight are more powerful than objective child body weight in predicting the development of eating disorders. Parent-perceived child overweight was also a specific risk factor for eating disorders, whereas elevated maternal weight and childhood psychosocial difficulties seem to be associated with increased risk for psychiatric disturbance more generally. These results have implications for the prevention of eating disorders, particularly in light of recent increases in the prevalence of childhood obesity. PMID- 19564800 TI - Six-month persistence of sleep problems in young children with autism, developmental delay, and typical development. AB - OBJECTIVE: This study examined the persistence of sleep problems in preschool children with autism and two matched comparison groups: children with developmental delay without autism and typically developing children. Sleep problems were defined subjectively by parent report, by the Children's Sleep Habits Questionnaire (CSHQ), and objectively by quantitative Research Diagnostic Criteria (RDC) derived from actigraphic recordings. METHOD: Children were studied on three occasions, each separated by a 3-month interval. At each assessment, the children were recorded actigraphically for 1 week, and parents completed sleep wake diaries and the CSHQ. Descriptive statistics and odds ratios were used to assess the occurrence and stability of sleep problems within children and across groups and to explore how actigraph- and CSHQ-defined sleep problems affect parental sleep problem reports. RESULTS: Parent reports of a generic sleep problem were more prevalent than RDC- and CSHQ-defined sleep problems, especially for children with neurodevelopmental disorders. For all groups, objectively measured sleep problems were rarely persistent during the 6-month period. The children in both neurodevelopmental groups, however, had more sleep problems on one or two occasions, using actigraph and the CSHQ, than typically developing children. CONCLUSIONS: Objective and subjective measures of sleep problems in preschool-aged children produce different results. In a community sample, the rate of actigraph- and CSHQ-defined sleep problems in children with autism did not differ from rates for typically developing children, although the parent report of a generic sleep problem was significantly greater. PMID- 19564801 TI - Sexual and drug use behavior in perinatally HIV-infected youth: mental health and family influences. AB - OBJECTIVE: As perinatally human immunodeficiency virus (HIV)-infected (PHIV+) youths enter adolescence, they are at high risk for poor behavioral and health outcomes. This study examines relations between youth mental health problems and sexual and substance use risk behavior, the impact of caregiver mental health and family functioning on youth mental health and risk behavior outcomes, and the role of youth HIV status in this process. METHOD: Participants were recruited from four medical centers. Individual interviews were administered to 193 PHIV+ and 127 perinatally HIV exposed but uninfected (PHIV-) 9- to 16-year-old boys and girls and their primary caregivers. Participants were primarily African American and Latino. The interview assessed child sexual and drug risk behavior, child and caregiver mental health, and family functioning. RESULTS: Exploratory latent variable structural equation modeling revealed no differences in rates of sexual risk behavior or substance use between PHIV+ and PHIV- youths. However, adolescent mental health was significantly associated with sexual risk behavior and substance use. Caregiver mental health was associated with youth mental health and indirectly with sexual risk behavior and drug use through its impact on youth mental health. Family functioning did not significantly predict youth outcomes. CONCLUSIONS: Over and above other key environmental factors and family functioning, youth and caregiver mental health problems are related to sex and drug use risk behaviors in PHIV+ and PHIV- youths. Given high rates of youth and caregiver mental health problems in this population, family-based mental health interventions may be a key component of HIV prevention programs for perinatally HIV-exposed youth. PMID- 19564802 TI - Ethnic differences in somatic symptom reporting in children with asthma and their parents. AB - OBJECTIVES: The purpose of this study is to examine the association between child and parent somatic symptom reporting and pediatric asthma morbidity in Latino and non-Latino white children. METHOD: The study consists of 786 children, 7 to 15 years of age, in Rhode Island (RI) and Puerto Rico. Children's and parents' levels of general somatic symptoms were assessed with well-established self report measures. Clinician-determined asthma severity was based on reported medication use, asthma symptom history, and spirometry results. Asthma-related health care use and functional morbidity was obtained via parent self-report. RESULTS: Child and parent reports of general somatic symptoms were significantly related to pediatric asthma functional morbidity when controlling for poverty, parent education, child's age, and asthma severity. In controlling for covariates, Latino children in RI reported higher levels of somatic symptoms than Island Puerto Rican children, and RI Latino parents reported more somatic symptoms than RI non-Latino white parents (p < .05). CONCLUSIONS: This study replicates and extends to children in previous research showing higher levels of symptom reporting in Latinos relative to whites. Results also provide new insight into the relation between general somatic symptom reports and pediatric asthma. Ethnic differences in somatic symptom reporting may be an important factor underlying asthma disparities between Latino and non-Latino white children. PMID- 19564803 TI - Sources of prescriptions for misuse by adolescents: differences in sex, ethnicity, and severity of misuse in a population-based study. AB - OBJECTIVE: Epidemiological data indicate that adolescent prescription misuse rates have risen over the past decade. Despite this, little work has examined sources for opioids, tranquilizers, and stimulants or evaluated sex or ethnic differences or whether different sources correspond to differences in other risk behaviors. METHOD: Data from the 2005 and 2006 National Survey on Drug Use and Health (adolescent n = 36,992) were used to address these questions. Frequencies and percentages for source categories were calculated, and potential sex and ethnic differences in medication source were evaluated using chi(2) analyses; logistic regression analyses evaluated whether the use of specific sources corresponded to a greater likelihood of concurrent substance use or depressive episodes. RESULTS: The most common source of medication was from friends or family, for free; other common sources included obtaining medication from a physician, purchasing medication, or theft (usually from friends or relatives). Sex differences were found, predominately for opioids: female patients were more likely to steal medication or obtain it for free; male patients were more likely to purchase medication or acquire it from a physician. White adolescents were more likely to purchase opioids, whereas African American adolescents were more likely to misuse opioids obtained from a physician. CONCLUSIONS: Across medication classes, adolescents who most recently acquired medication by purchasing it had the worst risk profile in terms of concurrent substance use and severity of prescription misuse. These results may help identify subgroups of adolescent prescription misusers who are most vulnerable to consequences from misuse or other substance use. PMID- 19564804 TI - Measuring the impact of menopausal symptoms on quality of life: methodological considerations. PMID- 19564805 TI - Estrogen for vulvovaginal symptoms: how low can you go? PMID- 19564806 TI - Backslab versus nonbackslab for immobilization of undisplaced supracondylar fractures: a randomized trial. AB - OBJECTIVES: Immobilization of supracondylar fractures of the humerus, for 1 to 4 weeks in a backslab or a collar and cuff, produces good functional outcomes. This study assesses the pain associated with these 2 forms of immobilization. METHODS: A randomized controlled trial comparing above-elbow posterior fiberglass slab and broad arm sling, with collar and cuff immobilization without plaster slab, was conducted at a children's hospital emergency department. Patients presenting with undisplaced supracondylar fractures were enrolled and reviewed after 2 weeks of immobilization. A daily diary was used during immobilization to assess pain, resumption of normal activity of the child, and parental satisfaction. Pain was assessed using a 100-mm visual analog scale. RESULTS: Fifty patients were enrolled, of whom 27 were randomized to a slab and 23 were randomized to a collar and cuff. Immobilization in a slab was associated with a reduction in time to resumption of normal activity, while immobilized in the device, with a median of 2.0 days (interquartile range [IQR], 1.0-5.0 days) versus 7.0 days (IQR, 3.0-13.0 days; P = 0.01). For the slab group, the median duration of pain was 4.0 days (IQR, 1-8.0 days) versus 6.0 days (IQR, 5.0-9.5 days) for the collar and cuff group (P = 0.07). Median daily pain severity was 28 mm for the slab group versus 33 mm for the collar and cuff group (P = 0.21). Moreover, 96% of parents in the slab group and 83% in the collar and cuff group would use the immobilization again (P = 0.17). CONCLUSIONS: For supracondylar fractures, use of an above-elbow posterior slab produces shorter duration of pain and reduces the time taken to become active again. PMID- 19564807 TI - Impact of an educational intervention on residents' knowledge of pediatric disaster medicine. AB - BACKGROUND: Globally, natural and created events have underscored the vulnerability of children in disasters. There is an unmet need for a standardized pediatric disaster medicine (PDM) curriculum. OBJECTIVE: To create and implement a PDM course, measure course efficacy, and assess residents' attitudes toward and experience in disaster medicine. DESIGN/METHODS: An educational intervention was conducted for pediatric and emergency medicine residents at a tertiary care teaching hospital. Participants completed a precourse survey of PDM attitudes and experience. Paired t tests were used to compare pretest, immediate posttest, and delayed posttest scores. Test performance was assessed by resident type and postgraduate year. A postcourse survey gauged reaction to the course and interest in further PDM training. RESULTS: Among the participants, 11 residents (9.4%) have treated disaster victims, and 5 (4.3%) had formal disaster medicine education. Most (83%) felt PDM is an important part of their training. Seventy five eligible residents (64.6%) completed the intervention. Pairwise comparison of scores showed a mean improvement in scores of 24.5% immediately after taking the course (95% confidence interval, 22.9%-30.1%; P < 0.001). Two months later, residents scored a mean of 69.0% for the delayed posttest, with a retained improvement in scores (18.3%; 95% confidence interval, 14.3%-22.3%; P < 0.001). Residents preferred future PDM exercises to additional didactic training (72.0% vs. 32.7%; P < 0.001%). CONCLUSIONS: Residents who complete this curriculum increase their knowledge of PDM with moderate retention of information. Most residents lack PDM training, believe it is important, and request disaster training exercises. PMID- 19564808 TI - A multidisciplinary approach to adverse drug events in pediatric trauma patients in an adult trauma center. AB - BACKGROUND: Adult trauma centers are major providers of medical management for pediatric trauma patients in the United States. Medication administration in this patient population is complex and fraught with potential error. METHODS: We designed a multidisciplinary team consisting of a pediatric hospitalist, pediatric care coordinator, pediatric nurse, pharmacist, and the trauma service to manage pediatric trauma patients from admission until discharge. The team mandated collective decision making for medication dosing and administration, weight documentation, and implemented a medication error reporting system. Our goal was to derive and implement a multidisciplinary practice and education-based model of pediatric trauma patient care to identify and decrease adverse medication events. RESULTS: Two hundred fifty-nine pediatric trauma patients were studied from January 1, 2003 to December 31, 2004, 125 pre-team implementation (control group) and 134 post-team implementation (study group). There were no significant differences in age, sex, mechanism of injury, injury severity score, or hospital length of stay between groups. There were significant reductions in number of medication prescribing errors (25 vs 15 errors; P = 0.05) and number of medication administration errors (19 vs 9 errors; P = 0.05) in the study group. Weight documentation improved significantly in the study group (90% vs 81%; P = 0.048). CONCLUSIONS: Instituting a multidisciplinary approach to pediatric trauma patient care is practical and can significantly decrease adverse medication events. PMID- 19564810 TI - Impact of a health literacy intervention on pediatric emergency department use. AB - OBJECTIVE: The aim of this study was to measure the impact of a simple parent health literacy intervention on emergency department and primary care clinic usage patterns. METHODS: Study participants consisted of parents who brought their children to the Harbor-UCLA Medical Center pediatric emergency department for nonurgent complaints. Study participants filled out questionnaires regarding their management of children's mild health complaints and where respondents first seek help when their children become sick. After completing the questionnaires, participants were educated about how to use the health aid book What to Do When Your Child Gets Sick and provided a free copy. After 6 months, telephone follow up interviews were conducted to assess whether the health literacy intervention had influenced the participants' management of their children's mild health complaints and their health care resource usage patterns. RESULTS: One hundred thirteen parents were enrolled in the preintervention phase, and 61 were successfully interviewed at 6 months by telephone. Before and after comparisons demonstrated a 13% reduction in the percentage of respondents who stated they would go to the emergency department first if their child became sick. In addition, 30% fewer respondents reported actual visits to the emergency department in the previous 6 months. Regarding specific low-acuity scenarios, significantly fewer participants would take their child to the emergency department for a low-grade fever with a temperature of 99.5 degrees F and for vomiting for 1 day. There was no significant change in the proportion of parents who would take their child to the emergency department for earache or cough. CONCLUSIONS: Health literacy interventions may reduce nonurgent emergency department visits and help mitigate emergency department overcrowding and the rising costs of health care. PMID- 19564809 TI - A randomized clinical trial of lidocaine gel for reducing infant distress during urethral catheterization. AB - OBJECTIVES: The purpose of this study was to determine whether a lidocaine enhanced lubricant that was used topically and instilled into the urethra decreased infants' distress that was associated with catheterization. METHODS: Eligible study participants were febrile 2- to 24-month-old pediatric emergency department patients in need of a catheter-obtained urinalysis. Patients were randomly assigned to 1 of 3 groups. Patients had catheterization performed after a 4-minute protocol that included a topical lubricant control, topical and intraurethral lubricants, or topical and intraurethral lidocaines. All patients were filmed during the procedure and evaluated at 3 phases: the start of the study, time of instillation, and at the time of maximal catheter insertion. The 3 phases were analyzed by blinded coders using the Modified Behavioral Pain Scale. RESULTS: A total of 45 patients were enrolled in 1 of the 3 study groups. Patients who received lidocaine had a lower overall distress at a level approaching significance at the time of catheterization (phase 3; P = 0.065) and a significantly lower distress as measured by crying during the catheterization (phase 3; P = 0.036) than infants who did not have a lubricant instilled into the urethra. CONCLUSIONS: The use of topical and intraurethral lidocaine resulted in a lower distress when compared with a topical lubricant, at a level approaching significance for global distress. When a subsection of the total distress score that measured infant cry was evaluated, the difference between the intraurethral lidocaine group and the group without intraurethral instillation was statistically significant. Although the results are promising, intraurethral lidocaine did not fully alleviate discomfort associated with urethral catherization. The use of lidocaine is suggested for pediatric patients undergoing urethral catherization; however, evaluation of additional agents and techniques for further decreasing pain is also warranted. PMID- 19564811 TI - Use of a safety resource center in a pediatric emergency department. AB - OBJECTIVE: To describe the use of a safety resource center (SRC) within a pediatric emergency department (ED) about injury prevention (IP) counseling, sales, costs, and parental responses to the services. METHODS: The SRC was established in June 2005 in the waiting area of an urban pediatric ED caring for approximately 96,000 patients annually. Safety resource center staff sells products of proven efficacy (eg, car seats, smoke alarms, and bike helmets) and offers safety education materials to patients and families. Activities including sales, educational content provided, types of inquiries from families, and overall satisfaction with the service were logged into a Microsoft Access database. Follow-up phone calls are made to all families 3 weeks after they purchase a product. RESULTS: Between June 20, 2005 and July 1, 2007, the SRC served approximately 13,000 families. Seven hundred eighty-six families purchased 816 products, generating $14,859. An additional 473 products were given away, totaling 1289 product items provided to families. The most commonly purchased items were car safety seats and bicycle helmets. Roughly 7000 IP-related brochures were distributed to ED families, and 120 car seats were fitted. Of the 786 families who made a purchase, 383 (49%) were reached for follow-up. Ninety seven percent reported to still be using the purchased product, and 28% made a different change in the home to practice safer behaviors. Ninety-five percent were grateful that the SRC was located in the ED. CONCLUSIONS: The SRC can provide IP product, encourage families to practice safer behaviors, and is well received within a large, urban pediatric ED. PMID- 19564812 TI - Efficacy of injectable trivalent virosomal-adjuvanted inactivated influenza vaccine in preventing acute otitis media in children with recurrent complicated or noncomplicated acute otitis media. AB - BACKGROUND: Most cases of acute otitis media (AOM) follow an upper respiratory infection due to viruses, including influenza viruses. As effective and safe influenza vaccines are available, their use has been considered among the possible measures of AOM prophylaxis. OBJECTIVES: To evaluate the efficacy of an inactivated virosomal-adjuvanted influenza vaccine in preventing AOM in children with a history of noncomplicated recurrent AOM (rAOM) or rAOM complicated by spontaneous perforation. METHODS: In this prospective, randomized, single blinded, placebo-controlled study, 180 children aged 1 to 5 years with a history of rAOM and previously unvaccinated against influenza were randomized to receive the inactivated virosomal-adjuvanted subunit influenza vaccine (n = 90) or no treatment (n = 90), and AOM-related morbidity was monitored every 4 to 6 weeks for 6 months. RESULTS: The number of children experiencing at least 1 AOM episode was significantly smaller in the vaccinated group (P < 0.001), as was the mean number of AOM episodes (P = 0.03), the mean number of AOM episodes without perforation (P < 0.001), and the mean number of antibiotic courses (P < 0.001); the mean duration of bilateral OME was significantly shorter (P = 0.03). The only factor that seemed to be associated with the significantly greater efficacy of influenza vaccine in preventing AOM was the absence of a history of recurrent perforation (crude odds ratio, P = 0.01; adjusted odds ratio, P = 0.006). CONCLUSIONS: The intramuscular administration of injectable trivalent inactivated virosomal-adjuvanted influenza vaccine in children with a history of rAOM significantly reduces AOM-related morbidity. However, the efficacy of this preventive measure seems to be reduced in children with rAOM associated with repeated tympanic membrane perforation. PMID- 19564813 TI - Target therapy for hepatocellular carcinoma: is sorafenib for everybody? PMID- 19564815 TI - Protection of intestinal permeability in the perioperative period. PMID- 19564814 TI - Predictors of gastric myoelectrical activity in type 2 diabetes mellitus. AB - BACKGROUND: Previous studies have clearly demonstrated the delayed gastric emptying of solid meals in diabetics, whereas their gastric myoelectrical activity, which primarily determines gastric motility, has not yet been fully confirmed. GOALS: This study aimed to clarify the characteristics and potential predictors of gastric myoelectrical activity in type 2 diabetics. STUDY: Twenty eight diabetics and 18 healthy controls participated. Duodenal biopsy sample was used for reverse transcription-polymerase chain reaction to evaluate cholecystokinin and motilin mRNA contents. Electrogastrography was performed before and after the test meal, and was assessed in terms of dominant frequency; dominant frequency instability coefficient; and the percentage of bradygastria, normogastria, and tachygastria. RESULTS: Over the entire recording period, dominant frequency was significantly lower, and dominant frequency instability coefficient and the percentage of bradygastria were significantly higher in diabetics than in controls. In diabetics, the multiple regression analysis demonstrated that dominant frequency instability coefficient and the percentage of tachygastria in the fasting period were dependent on fasting plasma glucose level and HbA1c, respectively. Moreover, dominant frequency over the entire period and the postprandial percentage of bradygastria were significantly associated with body mass index; the fasting percentage of bradygastria and postprandial dominant frequency instability coefficient were associated with fasting serum leptin level; the postprandial percentage of bradygastria was also associated with cholecystokinin mRNA content. CONCLUSIONS: Gastric myoelectrical activity in type 2 diabetics is impaired on dominant frequency, dominant frequency instability coefficient, and the percentage of bradygastria and predicted by body mass index, fasting serum leptin level, and cholecystokinin mRNA content besides the glycemic status. PMID- 19564816 TI - Effect of noise conditioning on cisplatin-induced ototoxicity: a pilot study. AB - BACKGROUND: Cisplatin is a platinum-based chemotherapeutic agent that is highly effective in the treatment of cancer. Ototoxicity is an important dose-limiting adverse effect of cisplatin, in addition to nephrotoxicity. Studies have shown that cisplatin-induced ototoxicity is mainly a result of generated reactive oxygen species. Sulfur-containing compounds such as L-N acetylcysteine (L-NAC) and D-methionine (D-MET) have shown promising results as potent otoprotectors against cisplatin-induced ototoxicity in animal studies. MATERIAL/METHODS: In this study, we investigated a method to increase the efficacy of L-NAC and D-MET without increasing dose. Sprague Dawley rats were noise conditioned for 15 minutes immediately after intraperitoneal injection of 275 mg/kg L-NAC or 300 mg/kg D-MET. Another set of rats received 275 mg/kg L-NAC or 300 mg/kg D-MET alone, and 1 group underwent noise conditioning alone. All 5 groups were administered 14 mg/kg cisplatin intravenously 1 hour after otoprotector injection or 45 minutes after noise conditioning. RESULTS: Otoprotectors and noise conditioning, alone or in combination, were analyzed for their ability to reduce cisplatin-induced ototoxicity. The results indicated that the combination of 275 mg/kg L-NAC and noise conditioning afforded more otoprotection than 275 mg/kg L NAC alone. In the case of D-MET, 300 mg/kg plus noise conditioning was little better than 300 mg/kg D-MET alone. In addition, we found that noise conditioning alone showed otoprotection against cisplatin-induced ototoxicity. CONCLUSIONS: The ability of noise conditioning to protect against cisplatin-induced ototoxicity requires additional study. PMID- 19564817 TI - Characterization of human and bovine phosphatidylethanolamine-binding protein (PEBP/RKIP) interactions with morphine and morphine-glucuronides determined by noncovalent mass spectrometry. AB - BACKGROUND: The phosphatidylethanolamine-binding protein (PEBP/RKIP), initially found to bind phosphatidylethanolamine (PE), has been shown to be associated with morphine derivatives. Our recent study on bovine primary chromaffin cells showed that inside secretory granules, PEBP is noncovalently associated to endogenous morphine-6-glucuronide (M6G), a highly analgesic morphine metabolite. During stress, M6G-PEBP complexes may be released into circulation to target peripheral opioid receptors. We now report the investigation of PEBP binding properties towards morphine and morphine analogs. MATERIAL/METHODS: Noncovalent electrospray ionization mass spectrometry (ESI-MS) was used to investigate bovine and human PEBP binding properties towards morphine and morphine-glucuronides. RESULTS: We describe for the first time that: (i) PEBP directly interacts with morphine glucuronides (M3G and M6G) but not with morphine, (ii) that the presence of a glucuronide group either on the 3rd or the 6th morphine's carbon does not affect these interactions, (iii) that M6G binds PEBP in a similar manner as the reference ligand PE and (iv) that PEBP displays a similar affinity for PE, M6G and M3G. CONCLUSIONS: Our results suggest that PEBP might protect M6G following its secretion into blood, leading to a longer half life. This study highlights the potentialities of ESI-MS to validate / invalidate the formation of protein: ligand noncovalent complexes when low affinity binders (i.e., compounds with affinities lower than 10(3) M(-1)) are concerned. PMID- 19564818 TI - The radioprotective effect of alpha2-macroglobulin: a morphological study of rat liver. AB - BACKGROUND: Administration of the acute-phase protein alpha2-macroglobulin (MG) prior to total-body irradiation of rats with a 6.7 Gy (LD50) dose of X-rays exerts a radioprotective effect. MATERIAL/METHODS: MG was administered 30 min before irradiation with a 6.7 Gy (LD50) dose of X-rays. Its radioprotective efficacy was compared with that of the synthetic agent amifostine (WR-2721), a sulfhydryl compound which is currently the most effective radioprotector in clinical use. After administration of either MG or amifostine, changes in body and liver weight were recorded and histological liver sections were examined during a four-week follow-up period. RESULTS: As observed in the experimental group administered amifostine, rats that received MG prior to irradiation exhibited 100% survival and restoration of the body and liver weight to the control values. The morphological damage seen in the liver after irradiation of untreated rats was absent in both the MG- and amifostine-pretreated rats. Also, hepatocytes and granulated cells had prominent nuclei and did not exhibit major changes in volume. Dilation of the central vein was not observed. CONCLUSIONS: Administration of MG before irradiation, similar to pretreatment with amifostine, provided complete survival of experimental rats and recovery of liver weight and preserved major histological parameters of the liver. PMID- 19564819 TI - Asthmatic bronchial fibroblasts demonstrate enhanced potential to differentiate into myofibroblasts in culture. AB - BACKGROUND: Chronic inflammation and remodeling of the bronchial wall are basic hallmarks of asthma. It is known that mesenchymal cells in the lamina reticularis underlying the basement membrane of the thickened airway wall of asthmatics predominantly display the phenotype of myofibroblasts and express alpha-smooth muscle actin (alpha-SMA). Human bronchial fibroblasts (HBFs) transform in vitro into myofibroblasts under the influence of transforming growth factor (TGF-beta). Differences in the reactivity of fibroblasts to TGF-beta in cultures derived from healthy and asthmatic donors are elucidated here. MATERIAL/METHODS: Primary human bronchial fibroblasts (HBFs) were cultured from bronchial biopsies from non asthmatic (n=7) and asthmatic (n=7) donors and treated with TGF-beta1 or TGF beta2 to induce myofibroblast differentiation. Expression of alpha-smooth muscle actin (alpha-SMA) was assessed by immunocytochemistry and Western blotting. The cell size and shape parameters were measured by computer-aided methods. RESULTS: Regardless of whether TGF-beta1 or TGF-beta2 was used, asthmatic cells showed enhanced expression of the myofibroblast marker as confirmed by immunocytochemistry and immunoblotting. Analysis of the shape parameters of cells incubated in the presence of TGF-beta1 revealed that HBFs of asthmatics differ from those of non-asthmatics. CONCLUSIONS: It is concluded that asthmatic HBFs cultured in vitro display some inherent features which facilitate their differentiation into myofibroblasts. These data indicate that increased reactivity of asthmatic fibroblasts to TGF-beta may play a crucial role in asthma. PMID- 19564820 TI - Prevalence of ESBL-positive bacteria in the community in the Czech Republic. AB - BACKGROUND: Problematic bacteria in the community include enterobacteria which produce extended-spectrum beta-lactamases. As yet there is no description of the prevalence of these bacteria in persons in the community in the Czech Republic. Therefore the main goal of this study was to determine the prevalence of ESBL positive enterobacteria in the gastrointestinal tracts of subjects in the community in the Czech Republic. MATERIAL/METHODS: Rectal swabs from the investigated subjects were inoculated onto chromID ESBL selective medium and enterobacteria were identified by the Vitek2 automated system. ESBL were detected using a modified DDST test. The results were confirmed by PCR and direct sequencing of CTX-M-positive amplicons. RESULTS: A total of 579 rectal swabs from subjects in the community were analyzed and ESBL production was both phenotypically and genotypically confirmed in 7 isolates. Thus the prevalence of ESBL-positive bacteria in the gastrointestinal tracts of the persons in the community was 1.2%. All the cases were Escherichia coli strains producing the CTX M-type ESBL. CTX-M-15 was the most prevalent type in this group of isolates. CONCLUSIONS: The presented results are in accord with other authors' studies and suggest that the epidemiologic profile of ESBL-positive enterobacteria in the Czech Republic is comparable to that in other European countries. PMID- 19564821 TI - The influence of age and diabetes on the skin blood flow response to local pressure. AB - BACKGROUND: Previous data has shown that when pressure is applied to the skin of the ankle and on the foot, there is a reactive increase in circulation. In the present investigation, these studies were expanded to look at the response of the hand, back, and foot to applied pressure. MATERIAL/METHODS: Ten young subjects whose average age was 26.5+/-3.3 yrs, 10 older subjects whose average age was 73.3+/-19.7 yrs and 10 people with diabetes whose average age was 60.1+/-5.7 yrs participated in the study. There was no statistical difference in the height or weight of the subjects. Hemoglobin A1c of the group with Diabetes averaged 6.98+/ 1.15% with the mean duration of diabetes 13.6+/-9.5 yrs. An infrared laser Doppler flow meter was used to measure circulation on the hand, lower back, and on the bottom of the foot during applications of pressure at 15, 30, 45, and 60 kPa. RESULTS: For all three areas of the body, circulation was significantly less in the group with diabetes than the other two groups (p<0.05). When pressure was applied at 15 kPa, the blood flow to the skin initially decreased, but then increased in the younger subjects and in the older subjects but did not increase in subjects with diabetes for any area of the body. Further, after pressure was released, for any of the four pressures examined here, while the younger subjects showed a pronounced reactive hyperemia, subjects with diabetes showed a diminished hyperemia not proportional to the pressure that was applied. CONCLUSIONS: It appears that the normal protective mechanism of a pressure induced hyperemia is absent or diminished in patients with diabetes with more effect on the periphery than on the core area of the body. More importantly, after pressure was applied and released, subjects with diabetes lacked a proportional hyperemia to recovery form the transient ischemia of the pressure. PMID- 19564822 TI - Hemodynamic relationships among upper-abdominal aorta and femoral arteries: basis for measurement of arterial blood flow to abdominal-pelvic organs. AB - BACKGROUND: Arterial blood flow (BF) to all abdominal-pelvic organs (AP) is a potentially useful indicator of splanchnic circulatory dysfunction or various stress-induced AP flow distributions. BFAP can be measured by subtracting BF in the "peripheral" bilateral limb left/right-femoral arteries (FA) from the "central" upper abdominal aorta (Ao) above the celiac trunk according to the formula: BFAP = BFAo-both BFFAs. It is necessary to understand the hemodynamics of the three-conduit arteries to determine physiological BFAP. The aim was to examine 1) basal hemodynamic parameters such as vessel diameter (VD), blood velocity (BV), and BF and their relationship to Ao and FAs and 2) how both BFAo and BFFAs influence the magnitude of BFAP. MATERIAL/METHODS: Fifty healthy males with a wide range of body weights were evaluated while seated following a 12-hour fast. Hemodynamics in three-conduit arteries was measured by pulsed Doppler with spectral analysis. RESULTS: Positive linear relationships in VD, BV, and BF were observed between Ao and both FAs (r value range: 0.399-0.529) and between BFAo and limb BFFAs (sum of both BFFAs; r=0.544, p<0.0001). BFAP was more strongly proportional to BFAo (r=0.966, p<0.0001) than BFFAs (r=0.303, p=0.0327 in LFA, r=0.281, p=0.0482 in RFA). BFAP was thus expressed as: BFAP (l/min) = 0.85 x BFAo 0.19. CONCLUSIONS: Under basal conditions, biological proportion and close hemodynamic relationship were observed between Ao and FAs. Consequently, BFAP was more strongly influenced by BFAo than by BFFAs. BFAP may be potentially evaluated by BFAo to determine single arterial hemodynamics using the above formula. PMID- 19564823 TI - Influence of genetic polymorphisms in oxidative stress related genes and smoking on plasma MDA-LDL, soluble CD40 ligand, E-selectin and soluble ICAM1 levels in patients with coronary artery disease. AB - BACKGROUND: Previous studies have shown that oxidative stress plays an important role in coronary heart disease. Polymorphisms in key enzymes that regulate oxidative stress may play a role in atherogenicity and were investigated in this study. MATERIAL/METHODS: One hundred and forty-three patients with angiographically proven coronary artery disease were studied. The effect of the C242T polymorphism of the p22phox gene, an essential component of the NADH/NADPH oxidase, and glutathione-S-transferase T1, M1 and P1 polymorphisms on plasma MDA LDL, soluble CD40 ligand, E-selectin and soluble ICAM1 levels was determined. Genotyping of the p22 phox C242T polymorphism was performed by RFLP analysis, and GSTT1, GSTM1 and GSTP1 genotypes were determined using a multiplex PCR assay. The MDA-LDL, sCD40L, E-selectin and sICAM1 levels were determined using ELISA. RESULTS: Patients with the TT or TC genotype of the p22 phox C242T polymorphism had significantly higher plasma MDA-LDL levels compared to those of the CC genotype. Plasma E-selectin and soluble ICAM1 levels were significantly higher in the TT or TC genotype compared to that of the CC genotype. In GSTT1+ patients, plasma MDA-LDL levels were significantly higher than those of GSTT1- patients. CONCLUSIONS: Genetic polymorphism of the p22 phox gene had a significant effect on plasma lipid peroxidation and endothelial function through oxidative stress. The results of this study confirm the effect of NADH/NADPH oxidase on atherogenecity. PMID- 19564824 TI - Brain wave P300: a comparative study of various forms of criminal activity. AB - BACKGROUND: This comparative and comprehensive study builds on a previous study comparing the P300 wave of impulsively violent delinquents and a non-impulsive non-delinquent group. The purpose was to investigate changes in P300 cognitive evoked potentials, especially the amplitude and latency at the Pz electrode site. MATERIAL/METHODS: The P300 parameters of perpetrators of various types of criminal offences and those of a control group matched for age, gender, and educational status were compared (N=80). There were 20 subjects with impulsively aggressive delinquent behavior. The observed parameters were compared with the neuropsychophysiological correlates of a group of 20 subjects with deliberately (i.e. non-impulsive) violent behavior, a group of 20 delinquents sentenced for property crimes (theft), and 20 non-delinquent non-impulsive nonviolent persons. To differentiate these groups, Eysenck's IVE questionnaire and a structured interview according to DSM IV criteria conducted by a certified forensic psychiatrist were used. RESULTS: The results showed a significantly lower P300 wave amplitude in the impulsively aggressive individuals than in the other groups. No significant differences were found in terms of latency. The results confirm the results of previous studies. CONCLUSIONS: The results suggest the possibility of a neuropsychophysiological correlate of impulsively aggressive individuals behaving in a socially dangerous way. This opens a discussion on the subject of expert evaluation of criminal acts within the context of "uncontrolled affect". PMID- 19564825 TI - Stuttering patients' opinions on the Digital Speech Aid. AB - BACKGROUND: The Digital Speech Aid (DSA) is a portable device used to reduce stuttering. It incorporates delayed auditory feedback (DAF) and frequency-shifted auditory feedback (FAF). Due to its small size, the DSA can be used by the stutterer in everyday life. MATERIAL/METHOD: Three hundred thirty-five stutterers aged 6-64 years were included in the study. A subgroup of 100 stutterers who had used the device for at least six months was asked to fill out a questionnaire of 25 questions about their opinion of the device. The data were analyzed and the factors determining the patients' satisfaction with the device were investigated. RESULTS: The analysis showed that the patients used the DSA regularly for about 3 hours daily. Most patients applied the device at home; only small number used it at work or at school. Most patients (70%) reported decreased fear of speaking and improved self-confidence during communication when using the device (78%). Eighty eight percent of the patients described the device as very useful or useful in everyday life. That patients' satisfaction with the device was not related to objective speech improvement but to a subjective feeling of increased self confidence and decreased fear of speaking. CONCLUSIONS: The DSA is positively judged by patients as an option in stuttering therapy. Its effect can be attributed not only to fluency improvement, but also to increased self-confidence and reduced fear of speaking. PMID- 19564826 TI - Primary lung cancer and TNF-alpha gene polymorphisms: a case-control study in a Croatian population. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is a multifunctional cytokine involved in the pathogenesis of various inflammatory and malignant diseases. Previous studies investigating the role of the TNF-alpha gene polymorphisms in lung cancer have generated contradictory results. The present study investigated whether the TNF-alpha-308 and TNF-alpha-238 polymorphisms are associated with risk and/or severity of disease in Croatian lung cancer patients. This is the first study in a Caucasian population to analyze the influence of these two polymorphisms on multiple types of lung cancer. MATERIAL/METHODS: In a case control study, lung cancer patients (n=230) and appropriate age- and sex-matched controls (n=230) were genotyped by the polymerase chain reaction/restriction fragment length polymorphism method. Allele and genotype frequencies were estimated by gene counting. The chi-squared test was used to compare the observed numbers of different TNF-alpha genotypes for the population with those predicted by Hardy-Weinberg equilibrium. Differences in genotype and allele distributions in the patient and control groups were analyzed for statistical significance using the chi-squared test or Fisher's exact test as appropriate. RESULTS: There were no significant differences in the genotype and allele frequencies for the TNF-alpha-308 and TNF-alpha-238 polymorphisms between lung cancer patients and controls. Furthermore, no association between the genotypes and different stages of lung cancer was detected. CONCLUSIONS: This study indicates that the TNF-alpha 308 and TNF-alpha-238 polymorphisms do not influence susceptibility to or severity of lung cancer in a Croatian population. PMID- 19564827 TI - Blood flow disturbances in the central retinal artery in patients with traumatic optic neuropathy. AB - BACKGROUND: Traumatic optic neuropathy (TON) is a serious sequel of head trauma, leading even to loss of vision. Experimental studies suggest that vascular factors contribute to the pathogenesis of TON. Thus the purpose of this study was to explore associations between blood flow Doppler parameters in the central retinal artery (CRA) and optic nerve function in patients after closed head trauma. MATERIAL/METHODS: Blood flow was evaluated by color-coded duplex sonography (CCDS) in the CRA of 72 consecutive patients (57 males and 15 females, mean age: 40+/-14.5, range: 15-77 years) with a diagnosis of traumatic optic neuropathy. Of the 101 affected eyes, optic nerve atrophy was found in 87 whereas the optic disc had normal appearance in the remaining 14 eyes. RESULTS: Both peak systolic velocity (PSV) and end-diastolic velocity (EDV) in the eyes with TON were significantly lower (26% and 20%, respectively, p<0.01) than those in age- and gender-matched healthy subjects. A statistically significant decrease in PSV was also noted in the CRA of the contralateral unaffected eye (9%, p<0.05). A statistically significant positive association was found between blood flow velocities in the CRA and visual acuity (p<0.01) after controlling for sex, side of symptoms, and blood pressure. CONCLUSIONS: Blood flow is significantly decreased in the central retinal artery of the affected eye and slightly in the unaffected eye in patients with traumatic optic neuropathy. Hemodynamic disturbances within the central retinal artery concur with functional nerve impairment. PMID- 19564828 TI - The importance of antibiotic prophylaxis in GBS-positive parturient women. AB - BACKGROUND: Bacterial streptococcal colonization of the vagina of a pregnant woman can be transferred to her newborn and induce a septic disease. The aim was to emphasize the requirements of timely intrapartal administration of antibiotics to prevent the transfer of GBS strains to the newborn. MATERIAL/METHODS: Three thousand two hundred seventy-eight pregnant women were evaluated who delivered babies during a 17-month period in 2006 and 2007 at the Department of Obstetrics and Gynecology in Ceske Budejovice. The results of their screening examinations from the 36th week of pregnancy were analyzed for information concerning their vaginal smears and smears from the newborns testing for GBS strains. RESULTS: The authors demonstrate that timely intrapartal administration of antibiotics considerably reduces the colonization of newborns with GBS strains (Fisher's exact test, p=0.0490). CONCLUSIONS: It is necessary to adhere consistently to preventive provisions which lead to a reduction in the colonization of newborns with GBS strains. Late application, i.e. within four hours before parturition, statistically significantly increases the risk of colonization with GBS. PMID- 19564829 TI - Positive impact of cyclic meditation on subsequent sleep. AB - BACKGROUND: Cyclic meditation (CM) is a technique that combines yoga postures interspersed with supine rest. This combination is based on ancient texts and is considered easier for beginners to practice. MATERIAL/METHODS: Whole-night polysomnographic measures and the self-rating of sleep were studied on the night following a day in which 30 male participants practiced CM twice (ca. 23 minutes each time). This was compared with another night when they had had two sessions of supine rest (SR) of equal duration on the preceding day. The sessions were one day apart and the order of the sessions was randomized. Recordings were from the F4, C4, and O2 electrode sites referenced to linked earlobes and bipolar electroculography and electromyography sites. RESULTS: In the night following CM, the percentage of slow-wave sleep (SWS) was significantly higher than in the night following SR, whereas the percentage of rapid-eye-movement (REM) sleep and the number of awakenings per hour were less. Following CM the self-rating of sleep based on visual analog scales showed an increase in the feeling that the sleep was refreshing, an increase in feeling "good" in the morning, an overall increase in sleep duration, and decreases in the degree to which sleep was influenced by being in a laboratory as well as any associated discomfort. CONCLUSIONS: Practicing cyclic meditation twice a day appeared to improve the objective and subjective quality of sleep on the following night. PMID- 19564830 TI - Unilateral developments of osteoarthritis and Charcot's joint in a patient with neurofibromatosis. AB - BACKGROUND: Precise mechanism of developing neuropathic arthropathy known as Charcot's joint is not fully understood. CASE REPORT: A 55-year-old Japanese woman with neurofibromatosis-1 complained of right gonalgia in December 2001. Physical examination revealed a huge tumor in the right lower leg without signs of inflammation. Laboratory findings were unremarkable. Radiographic examination disclosed the presence of osteoarthropathy in the right knee joint. In contrast, radiologic findings of the right foot and ankle were compatible with neuropathic arthropathy. Further investigations could not reveal abnormal findings in the nervous system. To improve patient's quality of life, partial resection of the tumor was performed. The resected tissues were compatible with neurofibromatosis without malignant transformation. The patient newly noticed pains in the right ankle and tarsal joints one year after the operation. Restricted mobility and insufficient blood supply in the right knee arising from the huge tumor might accelerate development of osteoarthropathy through malnutrition of the chondrocytes. Because the patient did not experience the arthralgia before the operation, the tumor might damage the peripheral nerves unabling to receive afferent signals from such joints resulting in neuropathic arthropathy. CONCLUSIONS: The damaged peripheral nerves might be contributory to developing or accelerating neuropathic arthropathy. PMID- 19564831 TI - Clinical findings of hibernoma of the buttock and thigh: rare involvements and extremely high uptake of FDG-PET. AB - BACKGROUND: Hibernoma is a rare adipose tissue tumor of the soft tissue and the term is derived from the histological similarities to the brown fat found in hibernating animals. It usually occurs in the interscapular area, back, and neck and a few cases have been reported in the buttock or thigh. CASE REPORT: Two cases are presented, one of which had a lesion in the buttock and the other a lesion in the thigh. The lesion in the buttock extended into the pelvis. CT and MR findings suggested liposarcoma in both cases, but the uptake was extremely intense on 18F-FDG-PET in one case. This was not typical of liposarcoma and suggestive of hibernoma. Biopsy specimens revealed a proliferation of adipose cells with vacuolated granular eosinophilic cytoplasm. No cellular atypia or mitotic figures were observed in either case. A marginal excision was performed in one case and an intralesional excision in the other. There were no signs of local recurrence at the final follow-up. CONCLUSIONS: While occurrences in the buttock or thigh are exceedingly rare, hibernoma should be included in the differential diagnosis of an adipose tissue tumor in the thigh, even though the imaging findings mimic liposarcoma. 18F-FDG-PET may be a very meaningful technique to differentiate hibernoma from liposarcoma. A correct diagnosis should be established to prevent over-surgery. PMID- 19564832 TI - Liposarcoma of the esophagus: case report and literature review. AB - BACKGROUND: Liposarcoma is one of the most common soft tissue sarcomas in adults, but it is exceedingly rare in the gastrointestinal tract. To the authors' knowledge, only 17 cases of esophageal liposarcomas have been described in the literature. CASE REPORT: The case of a 56-year-old woman is reported who complained of dyspnea and airway obstruction and was referred for multi-slice computed tomography (MSCT) with the suspicion of mediastinal tumor. MSCT revealed a heterogeneous mass within the esophagus. The density measurements from pre- and post-contrast scanning suggested a fatty tissue component of the lesion and a possible malignant nature. The patient underwent esophagotomy. The diagnosis of liposarcoma was established. Five months after the surgery, follow-up chest MSCT was performed which revealed an inhomogeneous mass behind the left atrium. Additional CT was performed with a perfusion protocol which suggested a malignant nature of the lesion. After thoracotomy with removal of the tumor, the patient underwent adjuvant radiotherapy. The patient is still being followed up and two years after the esophagotomy has been well, with no metastases in subsequent endoscopic and CT examinations. CONCLUSIONS: To the authors' knowledge this is the first case of liposarcoma of the esophagus detected and postoperatively monitored using MSCT. MSCT, even as the first examination, may enable a proper diagnosis of liposarcoma in patients with atypical clinical features. PMID- 19564833 TI - Prediction of hearing thresholds using the Cochlea-Scan. Comment to: A pilot study on assessing hearing threshold using the Cochlea-Scan. Stavros Hatzopoulos, Krzyszof Kochanek, Lech Sliwa, Henryk Skarzynski. Med Sci Monit, 2008; 14(4): MT7 11. PMID- 19564834 TI - Unnecessary diagnostic tests often obtained for benign paroxysmal positional vertigo. AB - BACKGROUND: Benign paroxysmal positional vertigo (BPPV) is a common cause of vertigo prompting patients to seek medical treatment. Diagnosis is made by identifying characteristic nystagmus with positional testing (e.g., Dix-Hallpike maneuver) during a clinical examination. Although diagnosis can be accurately made by non-specialists, costly diagnostic tests are frequently obtained. We examined diagnostic tests ordered and use of specialty consultations in the evaluation of patients eventually diagnosed in our clinic with BPPV. MATERIAL/METHODS: We retrospectively reviewed the medical records of 193 consecutive adult patients referred to our otoneurology clinic and subsequently diagnosed with isolated BPPV of the posterior canal from 2003 to 2008. In all cases, symptoms resolved after treatment. RESULTS: One hundred thirty-six patients (71%) underwent at least one test, and no tests were diagnostic. The test most frequently ordered was brain magnetic resonance imaging (n=76), followed by audiogram (n=64), serologic tests (n=42), brain computed tomographic scan (n=32), caloric electronystagmography (n=24), and magnetic resonance angiography (n=23). Internists and otolaryngologists evaluated the same number of patients (n=75) prior to being seen in our clinic, and neurologists saw the next largest group (n=52). Family and emergency physicians evaluated 22 and 16 patients, respectively. The number of diagnostic tests ordered did not significantly vary from 2003 to 2008. CONCLUSIONS: Patients with isolated BPPV undergo many tests of no yield despite the definitive test for diagnosis being a bedside maneuver. Increased use of tests for positional nystagmus likely would improve diagnostic yield and decrease costs in evaluating patients with this common disorder. PMID- 19564836 TI - Primary healthcare in the developing part of Europe: changes and development in the former Eastern Bloc countries that joined the European Union following 2004. AB - BACKGROUND: Primary care is an important tool to improve global health. This study presents an overview of how targets regarding primary care were realized in ten countries of the former Soviet Bloc that joined the EU since 2004. MATERIAL/METHODS: Demographic, socioeconomic, and mortality-based statistical data are presented, scientific publications from the countries analyzed, and personal experiences of family physicians of these countries compared. RESULTS: After the collapse of communism, political changes and healthcare reforms began in these countries. There was economic recession and decline in the first decade. Life expectancies improved and total health expenditures increased to different extents, although governments spent barely more for healthcare. Primary care providers are the main private-sector contributors. The hospital-based structure changed, while the number of outpatient contacts is nearly the same. The ratio between secondary care specialists and family physicians remains too high and there is a shortage of educated nurses. Although new funding systems for primary care were introduced, budgets were mostly redistributed without substantial increases or improvement in outcome. The achievements of reform have rarely been evaluated systematically. Teamwork and praxis communities do not exist. The old style of polyclinics still predominates in some countries. The gate-keeping system is often symbolic or dysfunctional. Health promotion and prevention are rarely supported. CONCLUSIONS: The implementation of family medicine is not an absolute priority for decision makers. The political situation is often unstable. Despite non-negligible achievements, the health systems in this part of Europe are still in the midst of transition. PMID- 19564835 TI - Residential care centers for persons with intellectual disability in Israel: trends in mortality from food choking during 1991-2004. AB - BACKGROUND: Unintentional injuries are the fifth leading cause of death in the general population, but even more of a concern in the population of persons with intellectual disability. The present study was conducted to investigate trends in choking resulting in death of persons with intellectual disability in residential care centers in Israel over the period of 1991-2004. MATERIAL/METHODS: Data for this study came from the Office of the Medical Director (OMD), Division for Mental Retardation (DMR), Ministry of Social Affairs and Social Services. Every case of death in residential centers is reported to the OMD, which has registered the place of residence, date of birth, age at death, sex, ethnic background, level of ID, cause of death, autopsy, and review committee, if this was established, in a data register since 1991. Data concerned with food choking were extracted from the mortality register for the 1991-2004 period. RESULTS: Over the study period (14 years) there was a total of 1019 cases of death with 27 (2.65%) caused by food choking. The average rate was 42.6 per 100,000 study population for food choking. CONCLUSIONS: The solution is prevention. In recent years the OMD together with other professionals have produced educational material and seminars on feeding and preventive measures. Not every case can be prevented, but it is important to identify risk factors, educate care givers, and ensure that food products that are common choking hazards are kept away from residents who are at risk. PMID- 19564837 TI - Antibody response to influenza vaccine in coronary artery disease: a substudy of the FLUCAD study. AB - BACKGROUND: Influenza vaccination is recommended for patients with cardiovascular diseases. The main reason for the low vaccination rates in such patients is insufficient knowledge about vaccination efficacy, including immune response to the vaccine. The aim of this study was to assess humoral response to influenza vaccination in patients with coronary artery disease. MATERIAL/METHODS: This was a substudy of the randomized prospective double-blind placebo-controlled FLUCAD study on influenza vaccination in the secondary prevention of ischemic coronary events in patients with coronary artery disease. Patients received inactivated subunit vaccine (n=325) or placebo (n=333). Anti-hemagglutinin and anti neuraminidase antibody levels to the vaccine strains as well as IgM and IgG levels against influenza A and B were measured before administration of vaccine/placebo and after 8-10 weeks in 78 vaccinated and 97 placebo patients. RESULTS: Post-vaccination antibody titers were significantly higher than before vaccination, with mean increases of 4.9- to 5.7-fold for anti-hemagglutinin and 3.5- to 4.2-fold for neuraminidase antibodies. Post-vaccination protection rates ranged from 56.4 to 60.3% and response rates from 62.8 to 68%. The percentage of patients with significant post-vaccination concentrations of IgG and IgM was higher than before vaccination and amounted to 100% and 88.1% in the case of IgG and 14.3% and 5.2% in the case of IgM in response to influenza A and B, respectively. CONCLUSIONS: At least 60% of the patients achieved high post vaccination antibody levels sufficient to prevent influenza. PMID- 19564838 TI - Influenza vaccination in the United States, 2005-2007. AB - BACKGROUND: This study presents current patterns of influenza vaccination among adults in the United States according to selected demographic factors, chronic diseases, and pregnancy status. MATERIAL/METHODS: Analyses are based on cross sectional survey data from the Behavior Risk Factor Surveillance System (BRFSS), 2005, 2006, and 2007. RESULTS: The percentage vaccinated with the flu shot significantly increased over the three study years (26%, 32%, and 37%, respectively). The percentage vaccinated using the flu spray remained near one. Levels of influenza vaccination significantly differed among racial/ethnic groups. Among men, the percentage vaccinated with the flu shot was 32% for whites, 27% for blacks, and 21% for Hispanics. Corresponding percentages for women were 37, 26, and 22. Flu vaccination also significantly increased with age, education, income, healthcare coverage, and presence of chronic disease. A higher percentage of pregnant women in the age range 18-44 were vaccinated than non pregnant women (24% vs. 20% with the flu shot; 0.7% vs. 1.0% with the flu spray). CONCLUSIONS: Influenza vaccination levels among adults remain lower than the Healthy People 2010 influenza vaccination objectives. PMID- 19564839 TI - The effects of perindopril on aortic elasticity and inflammatory markers in hypertensive patients. AB - BACKGROUND: Inflammation plays an important role in hypertension and in atherosclerosis. Inflammatory changes induced even in prehypertensive subjects can lead to increased arterial stiffness. The effects of perindopril on both inflammatory and aortic elasticity markers were tested in hypertensive patients. MATERIAL/METHODS: One hundred nine hypertensive patients not taking any antihypertensive therapy were included in the study. Aortic strain, aortic distensibility, aortic stiffness index, and inflammatory markers, including CRP, IL-1alpha IL-1beta, and TNF-alpha, were measured in all patients before and after twenty weeks of perindopril therapy. RESULTS: While aortic strain and distensibility showed statistically significant increases with perindopril therapy, the aortic stiffness index and inflammatory markers were found to decrease. CONCLUSIONS: Perindopril therapy resulted in an improvement in aortic elastic properties. There was also an attenuation of inflammatory status of the patients as reflected by lower inflammatory marker levels compared with pretreatment values. PMID- 19564840 TI - Acute pancreatitis: a literature review. AB - Acute pancreatitis (AP) is an inflammatory disease characterized by steady, acute abdominal pain of varying severity, often radiating from the epigastrium to the back. Its presentation ranges from a self-limiting mild disorder to a more severe and fulminant disease. Severe acute pancreatitis accounts for 30% of all deaths related to pancreatitis. The incidence of AP is increasing progressively with a corresponding increase in the incidence of its risk factors. Alcohol abuse and gallstone migration are the established risk factors for development of AP. In recent years, genetic factors and obesity have also been identified as risk factors responsible for the development of AP. The pathophysiology of AP involves acute inflammation of the acinar cells. Excessive acinar cell injury leads to a condition called systemic inflammatory response syndrome (SIRS). Protracted SIRS is responsible for most of the life-threatening complications associated with AP. Most common AP-related complications include pulmonary, renal, cardiovascular, and central nervous system dysfunction. Thus prompt and accurate diagnosis of AP is of paramount importance. The medical management of AP includes controlling pain, providing adequate nutritional support, and monitoring complications. Endoscopic retrograde cholangiopancreatography and surgery have also shown to reduce the mortality and morbidity associated with AP. Drugs such as resveratrol and rosiglitazone are being investigated as potential candidates for the treatment of AP. PMID- 19564841 TI - A review on the impact of systematic safety processes for the control of error in medicine. AB - Among risk management initiatives, systematic safety processes (SSPs), implemented within health care organizations, could be useful in managing patient safety. The purpose of this article is to conduct a systematic literature review assessing the impact of SSPs on different error categories. Articles that investigated the relation between SSPs, clinical and organizational outcomes were selected from scientific literature. The proportion and impact of proactive and reactive SSPs were calculated among five error categories. Proactive interventions impacted more positively than reactive ones in reducing medication errors, technical errors and errors due to personnel. PSSPs and RSSPs had similar effects in reducing errors related to a wrong procedure. A single reactive study influenced non-positively communication errors. A relevant prevalence of the impact of proactive processes on reactive ones is reported. This article can help decision makers in identifying which SSP can be the most appropriate against specific error categories. PMID- 19564842 TI - Focal Ca2+ transient detection in smooth muscle. AB - Ca2+ imaging of smooth muscle provides insight into cellular mechanisms that may not result in changes of membrane potential, such as the release of Ca2+ from internal stores, and allows multiple cells to be monitored simultaneously to assess, for example, coupling in syncytial tissue. Subcellular Ca2+ transients are common in smooth muscle, yet are difficult to measure accurately because of the problems caused by their stochastic occurrence, over an often wide field of view, in an organ that it prone to contract. To overcome this problem, we've developed a series of imaging protocols and analysis routines to acquire and then analyse, in an automated fashion, the frequency, location and amplitude of such events. While this approach may be applied in other contexts, our own work involves the detection of local purinergic Ca2+ transients for locating transmitter release with submicron resolution. ATP is released as a cotransmitter from autonomic nerves, where it binds to P2X1 receptors on the smooth muscle of the detrusor and vas deferens. Ca2+ enters the smooth muscle, resulting in purinergic neuroeffector Ca2+ transients (NCTs). The focal Ca2+ transients allow the optical monitoring of neurotransmitter release in a manner that has many advantages over electrophysiology. Apart from the greatly improved spatial resolution, optical recording has the additional advantage of allowing the recording of transmitter release from many distinguishable sites simultaneously. Furthermore, the optical plane of focus is easier to maintain or correct during long recording series than is the repositioning of an intracellular sharp microelectrode. In summary, a method for imaging of Ca2+ fluorescence is outlined which details the preparation of tissue, and the acquisition and analysis of data. We outline the use of several scripts for the analysis of such Ca2+ transients. PMID- 19564843 TI - Papillary hyperplasia of the gallbladder in pancreaticobiliary maljunction represents a senescence-related lesion induced by lysolecithin. AB - Cellular senescence, an irreversible growth arrest, is considered to play as safeguard against malignant progression, though such a mechanism is speculative in human carcinogenesis. In gallbladder carcinoma, cholecystolithiasis and pancreaticobiliary maljunction (PBM) are major risk factors. Here, by using 113 surgically resected gallbladders and cultures of human gallbladder epithelial cells (HGECs) and gallbladder carcinoma cell line (TGBC2TKB), we examined carcinogenesis with respect to cellular senescence. Among 15 cases of PBM in which carcinoma was found in 4 cases, nonneoplastic gallbladder mucosa showed diffuse papillary hyperplasia (PHP). PHP was not found in gallbladders with cholecystolithiasis. Interestingly, PHP exhibited senescent features such as expression of p16(INK4A) and low cell proliferative activity. In contrast, EZH2, a polycomb group protein, was overexpressed in intraepithelial neoplasm and carcinoma in gallbladders with cholecystolithiasis. In PBM, EZH2 was expressed only in carcinoma foci but not in PHP. Cultured HGECs treated with lysolecithin, the level of which is elevated in gallbladder bile of PBM, showed increased expression of p16(INK4A) and senescence-associated beta-galactosidase. Conversely, enforced overexpression of EZH2 in senescent HGECs reduced p16(INK4A) expression. A knockdown of EZH2 in cultured TGBC2TKB cells increased p16(INK4a) expression. In conclusion, PHP in PBM may act as a barrier to malignant transformation for decades. EZH2 may be responsible for the escape from cellular senescence followed by malignant transformation in the gallbladder of PBM. PMID- 19564844 TI - Adiponectin deficiency impairs liver regeneration through attenuating STAT3 phosphorylation in mice. AB - Liver regeneration is a very complex and well-orchestrated process associated with signaling cascades involving cytokines, growth factors, and metabolic pathways. Adiponectin is an adipocytokine secreted by mature adipocytes, and its receptors are widely distributed in many tissues, including the liver. Adiponectin has direct actions in the liver with prominent roles to improve hepatic insulin sensitivity, increase fatty acid oxidation, and decrease inflammation. To test the hypothesis that adiponectin is required for normal progress of liver regeneration, 2/3 partial hepatectomy (PH) was performed on wild-type and adiponectin-null mice. Compared to wild-type mice, adiponectin-null mice displayed decreased liver mass regrowth, impeded hepatocyte proliferation, and increased hepatic lipid accumulation. Gene expression analysis revealed that adiponectin regulated the gene transcription related to lipid metabolism. Furthermore, the suppressed hepatocyte proliferation was accompanied with reduced signal transducer and activator of transcription protein 3 (STAT3) activity and enhanced suppressor of cytokine signaling 3 (Socs3) transcription. In conclusion, adiponectin-null mice exhibit impaired liver regeneration and increased hepatic steatosis. Increased expression of Socs3 and subsequently reduced activation of STAT3 in adiponectin-null mice may contribute to the alteration of the liver regeneration capability and hepatic lipid metabolism after PH. PMID- 19564845 TI - Limitations and possibilities of small RNA digital gene expression profiling. PMID- 19564846 TI - RNAiCut: automated detection of significant genes from functional genomic screens. PMID- 19564847 TI - Enabling IMAC purification of low abundance recombinant proteins from E. coli lysates. PMID- 19564848 TI - Finding multiple needles in one immune haystack. PMID- 19564849 TI - Downhill protein folding under pressure. PMID- 19564852 TI - To eGFR or not to eGFR: here is an intern's answer. PMID- 19564854 TI - Urinary biomarkers: the future looks promising. AB - Soler-Garcia and collaborators show that changing levels of urinary biomarkers associated with the pathology of HIV-associated nephropathy may identify HIV infected children at greatest risk for this complication. These changes appear relatively early in disease progression and remain responsive to antiviral therapy. Will these trends be borne out in a multicenter study? PMID- 19564855 TI - Ghrelin and its analogues as therapeutic agents for anorexia and cachexia in end stage renal disease. AB - Anorexia and cachexia are prevalent in patients with end-stage renal disease (ESRD) and are associated with increased risks of morbidity and mortality. Ashby et al. provide evidence that subcutaneous administration of synthetic ghrelin stimulated food intake in ESRD patients over a week, with no subsequent reduction in energy intake and no significant adverse effects. However, the clinical utility of ghrelin will depend on long-term outcomes in improving appetite and lean body mass as well as morbidity and mortality. PMID- 19564856 TI - Interactions between angiotensin II and arginine vasopressin in water homeostasis. AB - Mice deficient in the angiotensin II type 1a (AT(1a)) receptor demonstrate a vasopressin-resistant nephrogenic diabetes insipidus. These knockout mice exhibit a threefold increase in 24-h urine excretion. Neither 2 weeks of exogenous vasopressin nor 5 days of fluid restriction reversed this polyuric state. This nephrogenic diabetes insipidus was associated with reductions in adenylyl cyclase protein and in the phosphorylated mitogen-activated protein kinase extracellular signal-regulated kinase 1/2. The results support an important interaction between vasopressin and angiotensin II in maximal urinary concentration. PMID- 19564857 TI - Autoantibodies against intercalated cells in Sjogren's syndrome. PMID- 19564858 TI - Comparison of antioxidant activity of cilnidipine and amlodipine. PMID- 19564860 TI - First use of nocturnal hemodialysis. PMID- 19564861 TI - The Case | A 66-year-old male with hyponatremia. Psychogenic polydipsia. PMID- 19564862 TI - Bladder augmentation using an autologous neo-bladder construct. PMID- 19564863 TI - An integrated funding mechanism for clinical trials in human gene therapy. PMID- 19564866 TI - A case for immunosuppression for myoblast transplantation in duchenne muscular dystrophy. PMID- 19564872 TI - Differential contribution of genetic variation in multiple brain nicotinic cholinergic receptors to nicotine dependence: recent progress and emerging open questions. AB - Nicotine dependence (ND), a major public health challenge, is a complex, multifactorial behavior, in which both genetic and environmental factors have a role. Brain nicotinic acetylcholine receptor (nAChR)-encoding genes are among the most prominent candidate genes studied in the context of ND, because of their biological relevance as binding sites for nicotine. Until recently, most research on the role of nAChRs in ND has focused on two of these genes (encoding the alpha4- and beta2-subunits) and not much attention has been paid to the possible contribution of the other nine brain nAChR subunit genes (alpha2-alpha3, alpha5 alpha7, alpha9-alpha10, beta3-beta4) to the pathophysiology and genetics of ND. This situation has changed dramatically in the last 2 years during which intensive research had addressed the issue, mainly from the genetics perspective, and has shown the importance of the CHRNA5-CHRNA3-CHRNB4 and CHRNA6-CHRNB3 loci in ND-related phenotypes. In this review, we highlight recent findings regarding the contribution of non-alpha4/beta2-subunit containing nAChRs to ND, based on several lines of evidence: (1) human genetics studies (including linkage analysis, candidate-gene association studies and whole-genome association studies) of several ND-related phenotypes; (2) differential pharmacological and biochemical properties of receptors containing these subunits; (3) evidence from genetically manipulated mice; and (4) the contribution of nAChR genes to ND related personality traits and neurocognitive profiles. Combining neurobiological genetic and behavioral perspectives, we suggest that genetic susceptibility to ND is not linked to one or two specific nAChR subtype genes but to several. In particular, the alpha3, alpha5-6 and beta3-4 nAChR subunit-encoding genes may play a much more pivotal role in the neurobiology and genetics of ND than was appreciated earlier. At the functional level, variants in these subunit genes (most likely regulatory) may have independent as well as interactive contributions to the ND phenotype spectrum. We address methodological challenges in the field, highlight open questions and suggest possible pathways for future research. PMID- 19564873 TI - Detrimental effects of glucocorticoids on neuronal migration during brain development. AB - Glucocorticoids, the most downstream effectors of the hypothalamus-pituitary adrenal axis, are one of main mediators of the stress reaction. Indeed, exposure to high levels of stress-triggered glucocorticoids is detrimental to brain development associated with abnormal behaviors in experimental animals and the risk of psychiatric disorders in humans. Despite the wealth of this knowledge, the cellular and molecular mechanisms underlying the detrimental effects of glucocorticoids on brain development remain unclear. Here, we show that excess glucocorticoids retard the radial migration of post-mitotic neurons during the development of the cerebral cortex, and identify an actin regulatory protein, caldesmon, as the glucocorticoids' main target. The upregulation of caldesmon expression is mediated by glucocorticoid receptor-dependent transcription of the CALD1 gene encoding caldesmon. This upregulated caldesmon negatively controls the function of myosin II, leading to changes in cell shape and migration. The depletion of caldesmon in vivo impairs radial migration. The overexpression of caldesmon also causes delayed radial migration during cortical development, mimicking the excessive glucocorticoid-induced retardation of radial migration. We conclude that an appropriate range of caldesmon expression is critical for radial migration, and that its overexpression induced by excess glucocorticoid retards radial migration during cortical development. Thus, this study provides a novel insight into the underlying mechanism of glucocorticoid-related neurodevelopmental disorders. PMID- 19564871 TI - Evolutionary analysis of the dynamics of viral infectious disease. AB - Many organisms that cause infectious diseases, particularly RNA viruses, mutate so rapidly that their evolutionary and ecological behaviours are inextricably linked. Consequently, aspects of the transmission and epidemiology of these pathogens are imprinted on the genetic diversity of their genomes. Large-scale empirical analyses of the evolutionary dynamics of important pathogens are now feasible owing to the increasing availability of pathogen sequence data and the development of new computational and statistical methods of analysis. In this Review, we outline the questions that can be answered using viral evolutionary analysis across a wide range of biological scales. PMID- 19564874 TI - Bipolar disorder with frequent mood episodes in the national comorbidity survey replication (NCS-R). AB - Virtually nothing is known about the epidemiology of rapid cycling bipolar disorder (BPD) in community samples. Nationally representative data are reported here for the prevalence and correlates of a surrogate measure of DSM-IV rapid cycling BPD from the National Comorbidity survey Replication (NCS-R), a national survey of the US household population. DSM-IV disorders were assessed in the NCS R with the WHO Composite International Diagnostic Interview (CIDI). Although the CIDI did not assess rapid cycling, it did assess the broader category of 12-month BPD with frequent mood episodes (FMEs), having at least four episodes of mania/hypomania or major depression in the 12 months before interview. Roughly one-third of NCS-R respondents with lifetime DSM-IV BPD and half with 12-month BPD met criteria for FME. FME was associated with younger age-of-onset (of BP-I, but not BP-II) and higher annual persistence (73% of the years since first onset of illness with an episode) than non-FME BPD. No substantial associations of FME vs non-FME BPD were found with socio-demographics, childhood risk factors (parental mental disorders, other childhood adversities) or comorbid DSM-IV disorders. However, FME manic episodes had greater clinical severity than non-FME episodes (assessed with a fully structured version of the Young Mania Rating Scale) and FME hypomanic episodes had greater role impairment than non-FME episodes (assessed with the Sheehan Disability Scales). Whether these indicators of severity merely reflect attenuated effects of rapid cycling or independent effects of sub-threshold rapid cycling warrants further study given the high proportion of lifetime cases who met criteria for FME. PMID- 19564876 TI - Maternal influence of prolyl endopeptidase on fat mass of adult progeny. AB - BACKGROUND: Maternal genotype has lifetime effects on progeny, but few specific genes, and no proteases, are known to underlie maternal effects. Prolyl endopeptidase (PREP) is a serine protease with putative substrates that regulate appetite or milk production. OBJECTIVE: To test effects of PREP on obesity phenotypes in mice. DESIGN: Mice with a gene trap (GT) of PREP (PREP(gt/gt)) on the C57BL/6J (B6) background were generated. Minimal PREP protein was detected by western blot. In Experiment 1, direct effects of PREP were measured in littermate mice derived from intercrosses of heterozygotes (PREP(WT/gt)). In Experiment 2, maternal effects of PREP were measured in reciprocal crosses of heterozygous (PREP(WT/gt)) and wild-type (WT) (PREP(WT/WT)) males and females. DIETS: Mice were fed either low-fat (LF, Experiments 1 and 2) or high-fat (HF, Experiment 1) defined diets. MEASUREMENTS: Adiposity index (AI) was calculated from body weight (BW) and weights of four fat depots measured in 120-day-old mice. Fasting plasma glucose, insulin and leptin were measured. In vivo plasma alpha-MSH levels were measured by targeted quantitative peptidomics. RESULTS: Experiment 1-In intercross mice, there were significant diet effects, but few genotype effects. There were no genotype effects on BW or AI in males or females on either diet. Experiment 2-In contrast, reciprocal crosses of heterozygous males or females with WT B6 revealed highly significant parent of origin effects on all traits except body length. Progeny (WT and heterozygous genotypes and both sexes) born to female PREP(WT/gt) heterozygotes had fat pads that weighed as much as -twofold more at 120 days old than progeny born to male heterozygotes. CONCLUSION: Heterozygosity for PREP GT results in highly significant maternal effects, whereas homozygosity for the PREP(gt/gt) mutation has a much more limited direct effect. PMID- 19564875 TI - Depot-specific differences in inflammatory mediators and a role for NK cells and IFN-gamma in inflammation in human adipose tissue. AB - BACKGROUND: Adipose tissue is a primary in vivo site of inflammation in obesity. Excess visceral adipose tissue (VAT), when compared to subcutaneous adipose tissue (SAT), imparts an increased risk of obesity-related comorbidities and mortality, and exhibits differences in inflammation. Defining depot-specific differences in inflammatory function may reveal underlying mechanisms of adipose tissue-based inflammation. METHODS: Stromovascular cell fractions (SVFs) from VAT and SAT from obese humans undergoing bariatric surgery were studied in an in vitro culture system with transcriptional profiling, flow cytometric phenotyping, enzyme-linked immunosorbent assay and intracellular cytokine staining. RESULTS: Transcriptional profiling of SVF revealed differences in inflammatory transcript levels in VAT relative to SAT, including elevated interferon-gamma (IFN-gamma) transcript levels. VAT demonstrated a broad leukocytosis relative to SAT that included macrophages, T cells and natural killer (NK) cells. IFN-gamma induced a proinflammatory cytokine expression pattern in SVF and adipose tissue macrophages (ATM). NK cells, which constitutively expressed IFN-gamma, were present at higher frequency in VAT relative to SAT. Both T and NK cells from SVF expressed IFN gamma on activation, which was associated with tumor necrosis factor-alpha expression in macrophages. CONCLUSION: These data suggest involvement of NK cells and IFN-gamma in regulating ATM phenotype and function in human obesity and a potential mechanism for the adverse physiologic effects of VAT. PMID- 19564877 TI - A randomized double-blind placebo-controlled study of the long-term efficacy and safety of diethylpropion in the treatment of obese subjects. AB - OBJECTIVE: To evaluate the efficacy of diethylpropion on a long-term basis, with emphasis in cardiovascular and psychiatric safety aspects. DESIGN: Randomized, double-blind, placebo-controlled trial. MEASUREMENTS: Following a 2-week screening period, 69 obese healthy adults received a hypocaloric diet and were randomized to diethylpropion 50 mg BID (n=37) or placebo (n=32) for 6 months. After this period, all participants received diethylpropion in an open-label extension for an additional 6 months. The primary outcome was percentage change in body weight. Electrocardiogram (ECG), echocardiography and clinical chemistry were performed at baseline and every 6 months. Psychiatric evaluation and application of Hamilton rating scales for depression and anxiety were also performed by experienced psychiatrists at baseline and every 3 months. RESULTS: After 6 months, the diethylpropion group lost an average of 9.8% (s.d. 6.9%) of initial body weight vs 3.2% (3.7%) in the placebo group (P<0.0001). From baseline to month 12, the mean weight loss produced by diethylpropion was 10.6% (8.3%). Participants in the placebo group who were switched to diethylpropion after 6 months lost an average of 7.0% (7.7%) of initial body weight. The difference between groups at month 12 was not significant (P=0.07). No differences in blood pressure, pulse rate, ECG and psychiatric evaluation were observed. Dry mouth and insomnia were the most frequent adverse events. CONCLUSION: Diethylpropion plus diet produced sustained and clinically significant weight loss over 1 year. It seems to be safe in relation to cardiovascular and psychiatric aspects in a well selected population. PMID- 19564878 TI - Prevalence of sarcopenia and sarcopenic obesity in Korean adults: the Korean sarcopenic obesity study. AB - OBJECTIVES: To examine the prevalence of sarcopenia and sarcopenic obesity (SO) as defined by different indices, including appendicular skeletal muscle mass (ASM)/height(2), skeletal muscle mass index (SMI) and residuals for Korean adults, and to explore the association between SO and metabolic syndrome. METHODS: Our study sample included 526 participants (328 women, 198 men) for whom complete data on body composition were collected using available dual X-ray absorptiometry. Modified National Cholesterol Education Program Adult Treatment Panel III criteria were used to identify the individuals with metabolic syndrome. RESULTS: The prevalence of sarcopenia and SO is higher in older adults. Using two s.d. of ASM/height(2) below reference values from young, healthy adults as a definition of sarcopenia, the prevalence of sarcopenia and SO was 6.3% and 1.3% in older (> or =60 years) men and 4.1% and 0.8% in older women, respectively. The prevalence of sarcopenia using the residuals method was 15.4% in older men and 22.3% in older women. In addition, using two s.d. of SMI, the prevalence of sarcopenia and SO was 5.1% and 5.1%, respectively, in older men and 14.2% and 12.5%, respectively, in older women. Among women, SO subjects defined by the SMI had three times the risk of metabolic syndrome (odds ratios (OR)=3.24, 95% confidence interval (CI)=1.21-8.66) and non-sarcopenic obese subjects had approximately twice the risk of metabolic syndrome (OR=2.17, 95% CI=1.22-3.88) compared with normal subjects. Similar trends were observed in men. CONCLUSION: The prevalence and cutoff values of sarcopenia and SO in the Korean population were evaluated using different methods. Among the different indices of sarcopenia and SO, SO only defined using the SMI was associated with the risk of metabolic syndrome. As the Korean population gets older and more obese, the problematics of SO need to be elucidated. PMID- 19564879 TI - BMI peak in infancy as a predictor for later BMI in the Uppsala Family Study. AB - BACKGROUND: The development of obesity through childhood, often characterized by using body mass index (BMI), has received much recent interest because of the rapidly increasing levels of obesity worldwide. However, the extent to which the BMI trajectory in the first year of life (the BMI 'peak' in particular) is associated with BMI in later childhood has received little attention. SUBJECTS: The Uppsala Family Study includes 602 families, comprising mother, father and two consecutive singleton offspring, both of whom were delivered at the Uppsala Academic Hospital, Sweden, between 1987 and 1995. The children's postnatal growth data, including serial measurements of height and weight (from which BMI was calculated), were obtained from health records. All children had a physical examination when they were aged between 5 and 13 years, at which height and weight were again recorded and used to calculate age- and sex-adjusted BMI z scores. METHODS: Subject-specific growth curves were fitted to the infant BMI data using penalized splines with random coefficients, and from these the location of the BMI peak for each participant was estimated. A multilevel modelling approach was used to assess the relationships between the BMI peak and BMI z-score in later childhood. RESULTS: The BMI peak occurred, on average, slightly later in female children, with a higher BMI peak in male children. Considered separately, both age and BMI at BMI peak were positively associated with later BMI z-score. Considered jointly, both dimensions of BMI peak retained their positive associations. CONCLUSIONS: The growth trajectory associated with higher childhood BMI appears to include a later and/or higher BMI peak in infancy. PMID- 19564880 TI - Fetal macrosomia and adolescence obesity: results from a longitudinal cohort study. AB - OBJECTIVE: To assess the association between fetal macrosomia and adolescent obesity. DESIGN: Longitudinal cohort study of the association between macrosomia and adolescent obesity. SUBJECTS: Between 1 October 2005 and 1 February 2007, a follow-up study of live-born infants born in 1993-1995 in Wuxi, a suburban area of Shanghai, was conducted. Subjects with birth weight > 4000 g were selected as the exposed. For each exposed subject, one subject with a birth weight of 2500 4000 g, matched by year of birth, sex of infant, and type of institute at birth, was chosen as non-exposed. Clinical data were collected by structured interview and physical examination. Obesity was defined as body mass index (weight (kg)/height (m(2))) higher than the sex-age-specific criteria by the working group on obesity in China. Distribution of baseline characteristics and adolescent obesity rate between the exposed and non-exposed groups was compared. RESULTS: A total of 1435 pairs of exposed and non-exposed subjects were included in the final analysis. No major difference in baseline characteristics (other than birth weight) was found between the exposed and non-exposed groups. Obesity rate was significantly higher in the exposed group (2.9%) than in the non-exposed group (1.6%). Adolescent obesity rates were 1.4, 1.9, 2.6, and 5.6%, respectively, in study subjects with a birth weight of 2500-3499, 3500-3999, 4000 4499, and > or =4500 g. The association between birth weight and adolescent obesity remained essentially the same when mother's demographic and anthropometric factors, breast feeding, and adolescent life-style factors were adjusted. CONCLUSION: Compared with infants of normal birth weight, infants with birth weight >4000 g, especially those >4500 g, are at increased risk of adolescent obesity. PMID- 19564881 TI - Percutaneous enterogastric tube dislodgement in tetraplegics. AB - STUDY DESIGN: Case reports describing abdominal complications following percutaneous enteral gastric (PEG) tube placement in three tetraplegics. OBJECTIVES: The aim was to increase provider awareness of PEG tube dislodgement as a complication in tetraplegics. SETTING: Spinal cord injury center, Veteran's Administration hospital. METHODS: The charts of three spinal cord injured veterans with tetraplegia were reviewed for clinical presentation and radiographic findings supporting the diagnosis of acute abdominal complications following PEG tube dislodgement. RESULTS: PEG tube dislodgement in tetraplegics can present with occult findings and lead to disastrous outcomes if the diagnosis is delayed. CONCLUSION: PEG tube dislodgement should be considered in tetraplegics who develop even subtle abdominal complaints, especially if the injury is complete. Further assessment with computed tomography (CT) scans or fistulograms should be considered to help with decision making. PMID- 19564882 TI - Sensory anal examination in spinal cord injury. PMID- 19564883 TI - Occupation in spinal cord injury patients in Turkey. AB - STUDY DESIGN: A cross-sectional survey. OBJECTIVES: To investigate the changes in the occupation of patients after spinal cord injury (SCI) and the factors that cause this change. SUBJECTS: The study involved 192 Turkish patients (41 female, 151 male) who had suffered SCI. The mean age of patients was 36.1+/-12.0 years. The mean follow-up time was 43.4+/-38.0 months. Before injury, 138 patients were employed in gainful occupations, 26 patients were housewives, 10 were retired, 7 were students and 11 patients were unemployed. Only 15 patients (7.8%) returned to their original occupations after injury. Thirteen patients (6.8%) are currently working in another job; 1 patient (0.5%) is a student; 10 (5.2%) are retired as was earlier; 40 (20.8%) are retired on grounds of disability; 26 (13.5%) are housewives; and 87 patients (45.3%) are currently unemployed. METHODS: Prospective data collection through a face-to-face interview on an established SCI Turkish sample. RESULTS: In our study, the rate of returning to work was found to be 14.6%. In the evaluation of factors affecting return to work after injury, educational level (P=0.00), pre-injury employment (P=0.01) and bladder-emptying method (P=0.03) were statistically significantly correlated with return to work. CONCLUSION: In this study, education, pre-injury employment and bladder-emptying method were found to be important factors in returning to work after SCI. PMID- 19564885 TI - Imaging outcomes in cardiovascular clinical trials. AB - Imaging has had an important role in cardiovascular disease over the past decade, with the increasing reliance on imaging outcomes as surrogates for clinical end points. Clinical trials now show a trend towards the use of functional, rather than anatomical, imaging modalities. Use of these powerful tools needs to be optimized in the design of cardiovascular trials. In the future, imaging modalities will be fundamental to research and drug development and an increased emphasis will be placed on the relationship between the results of imaging studies and clinical outcomes. PMID- 19564884 TI - Gender differences in the cardiovascular effect of sex hormones. AB - The higher incidence of cardiovascular disease in men than in women of similar age, and the menopause-associated increase in cardiovascular disease in women, has led to speculation that gender-related differences in sex hormones have a key role in the development and evolution of cardiovascular disease. Compelling data have indicated that sex differences in vascular biology are determined not only by gender-related differences in sex steroid levels, but also by gender-specific tissue and cellular differences that mediate sex-specific responses. In this Review, we describe the sex-specific effects of estrogen and testosterone on cardiovascular risk, direct vascular effects of these sex hormones, and how these effects influence development of atherosclerosis. Cardiovascular effects of exogenous hormone administration are also discussed. Importantly, evidence has indicated that estrogens alone or in combination with progestins in postmenopausal women increase cardiovascular risk if started late after menopause, but that it possibly has beneficial cardiovascular effects in younger postmenopausal women, although data on long-term testosterone therapy are lacking. Hormone therapy should not be considered solely for primary prevention or treatment of cardiovascular disease at this time. PMID- 19564886 TI - Genomics of type 2 diabetes mellitus: implications for the clinician. AB - Our understanding of the genetics of type 2 diabetes mellitus (T2DM) has changed, in part owing to implementation of genome-wide association studies as a method for unraveling the genetic architecture of complex traits. These studies enable a global search throughout the nuclear genome for variants that are associated with specific phenotypes. Currently, single nucleotide polymorphisms in about 24 different genetic loci have been associated with T2DM. Most of these genetic loci are associated with the insulin secretion pathway rather than insulin resistance. Study design, heritability differences and the intrinsic properties of in vivo insulin resistance measures might partially explain why only a few loci associated with insulin resistance have been detected through genome-wide association approaches. Despite the success of these approaches at detecting loci associated with T2DM, currently known associations explain only a small amount of the genetic variance involved in the disease. Compared with previous studies, larger cohorts might be needed to identify variants of smaller effect sizes and lower allele frequencies. Finally, the current list of genetic loci that are related to T2DM does not seem to offer greater predictive value in determining diabetes risk than do commonly used phenotypic risk factors and family history. PMID- 19564888 TI - The roles of interferon-gamma and perforin in antiviral immunity in mice that differ in genetically determined NK-cell-mediated antiviral activity. AB - The design of effective antiviral immunotherapies depends on a detailed understanding of the cellular and molecular processes involved in generating and maintaining immune responses. Control of cytomegalovirus (CMV) infection requires the concerted activities of both innate and adaptive immune effectors. In the mouse, immunity to acute murine CMV (MCMV) infection depends on natural killer (NK) cells and/or CD8(+) T cells. The relative importance of NK and CD8(+) T cells varies in different mouse strains. In C57BL/6 mice, early viral infection is controlled by Ly49H(+) NK cells, whereas in BALB/c mice, CD8(+) T cells exert the principal antiviral activities. Although the role of NK and CD8(+) T cells is defined, the molecular mechanisms they utilize to limit acute infection are poorly understood. Here, we define the specific roles of perforin (pfp) and interferon-gamma (IFN-gamma) in the context of NK- or T-cell-mediated immunity to MCMV during acute infection. We show that pfp is essential for both NK- and T cell-mediated antiviral immunity during the early stages of infection. The relative importance of IFN-gamma is more pronounced in Ly49H(-) mice. Using BALB/c background mice congenic for Ly49H and lacking pfp, we show that Ly49H regulated NK-cell control of MCMV infection is dependent on pfp-mediated cytolysis. PMID- 19564887 TI - Familial pituitary tumor syndromes. AB - The vast majority of pituitary tumors are benign and occur sporadically; however, they can still result in significant morbidity and even premature mortality through mass effects and hormone dysfunction. The etiology of sporadic tumors is still poorly understood; by contrast, advances have been made in our understanding of familial pituitary adenoma syndromes in the past decade. Currently, four genes are known to be associated with familial pituitary tumor syndromes: MEN1, CDKN1B, PRKAR1A and AIP. The first three genes are associated with a variety of extrapituitary pathologies, for example, primary hyperparathyroidism with multiple endocrine neoplasia type 1, which might aid identification of these syndromes. By contrast, AIP mutations seem to occur in the setting of isolated familial pituitary adenomas, particularly of the growth hormone-secreting subtype. Awareness and identification of familial pituitary tumor syndromes is important because of potential associated pathologies and important implications for family members. Here, we review the current knowledge of familial pituitary tumor syndromes. PMID- 19564890 TI - Cross-regulation of the Nanog and Cdx2 promoters. AB - The first cell fate choice in the mammalian embryo, the segregation of the inner cell mass (ICM) and trophectoderm (TE), is regulated by the mutually antagonistic effects of the transcription factors, Oct4 and Cdx2, while the pluripotency factor, Nanog, is essential to specify the epiblast. We have analyzed the promoters of Nanog and Cdx2, and have found that these two transcription factors are likewise regulated reciprocally. Using an embryonic stem cell line with conditional TE differentiation, we show that Nanog overexpression suppresses the upregulation of TE markers, while Nanog knockdown upregulates the expression of TE markers. We further show that Nanog and Cdx2 bind to and repress each other's promoters. However, whereas Nanog knockout results in detectable Cdx2 expression in the ICM, we observe no overt disruption of blastocyst development, indicating that Nanog plays a subservient role to Oct4 in segregation of the ICM and TE. PMID- 19564891 TI - Enhancement of TRAIL cytotoxicity by AG-490 in human ALL cells is characterized by downregulation of cIAP-1 and cIAP-2 through inhibition of Jak2/Stat3. AB - The ability of death-inducing tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to selectively kill a variety of cancer cells has been largely described, but one of the major concerns with the treatment is the occurrence of drug resistance and possible toxic side effects. Here, we report that TRAIL induces apoptosis in Jurkat and SUPT1 T cell lines and in human T-ALL blasts but not in healthy subject-derived peripheral blood mononuclear cells. In parallel, the treatment with TRAIL and Tyrphostin (AG-490), a selective Janus kinase 2 inhibitor, produces an evident enhancement of cytotoxicity, characterized by a significant inhibition of Stat3 phosphorylation compared to controls or to TRAIL alone-treated samples, and associated with a dramatic decrease of both cIAP-1 and cIAP-2 mRNA levels. Downregulation of cIAP-1 and cIAP-2 by specific small interference RNAs significantly amplifies TRAIL-reduced cytotoxicity. All together, these findings strongly indicate that cIAP-1 and cIAP-2 downregulation is a fundamental step in the signaling pathways mediating the combinatorial effect of TRAIL and AG-490 on T cell leukemia. These findings may help to open new routes for the development of less toxic pharmacological strategies in the treatment of patients affected by TRAIL-sensitive leukemias. PMID- 19564889 TI - Chemotherapy-associated renal dysfunction. AB - The presence of renal dysfunction in a patient receiving chemotherapy can be devastating. Although many patients with cancer have underlying compromised renal function, some chemotherapeutic agents can actually induce renal abnormalities. An understanding of which traditional and newer chemotherapy agents can affect renal function is useful for physicians so that they can monitor patients for renal abnormalities and initiate preventive strategies to minimize renal complications. This Review highlights renal abnormalities associated with current chemotherapy agents and provides suggestions for preventive measures. PMID- 19564892 TI - CDP1, a novel component of chloroplast division site positioning system in Arabidopsis. AB - Chloroplasts are plant-specific organelles that evolved from endosymbiotic cyanobacteria. They divide through binary fission. Selection of the chloroplast division site is pivotal for the symmetric chloroplast division. In E. coli, positioning of the division site at the midpoint of the cell is regulated by dynamic oscillation of the Min system, which includes MinC, MinD and MinE. Homologs of MinD and MinE in plants are involved in chloroplast division. The homolog of MinC still has not been identified in higher plants. However, an FtsZ like protein, ARC3, was found to be involved in chloroplast division site positioning. Here, we report that chloroplast division site positioning 1 (AtCDP1) is a novel chloroplast division protein involved in chloroplast division site placement in Arabidopsis. AtCDP1 was discovered by screening an Arabidopsis cDNA expression library in bacteria for colonies with a cell division phenotype. AtCDP1 is exclusively expressed in young green tissues in Arabidopsis. Elongated chloroplasts with multiple division sites were observed in the loss-of-function cdp1 mutant. Overexpression of AtCDP1 caused a chloroplast division phenotype too. Protein interaction assays suggested that AtCDP1 may mediate the chloroplast division site positioning through the interaction with ARC3. Overall, our results indicate that AtCDP1 is a novel component of the chloroplast division site positioning system, and the working mechanism of this system is different from that of the traditional MinCDE system in prokaryotic cells. PMID- 19564893 TI - Trichosanthin functions as Th2-type adjuvant in induction of allergic airway inflammation. AB - It is important to understand the pathogenesis of asthma induced by natural allergens, which could exclude the interference of artificial adjuvant and provide insights of natural immune response in the disease. In the present study, we show that Trichosanthin (TCS) could induce airway inflammation even without the help of alum. Furthermore, TCS appeared capable of replacing alum to promote OVA-specific airway inflammation. TCS induced accumulation of IL-4-producing eosinophils in peritoneum at an early stage and the adjuvant function of TCS was eliminated by blockage of IL-4 at this stage. Finally, the eosinophils triggered by TCS from WT mice, but not from IL-4-deficient mice were shown to function as adjuvant for the induction of OVA-specific Th2 responses. Our data indicate that TCS is not only an allergen, but also a Th2-type adjuvant modulating the switching of immune responses to a Th2 pathway. This chain of events results from IL-4 production by eosinophils at an early stage of TCS-priming. In conclusion, TCS may be useful as a Th2 adjuvant, and innate immune cells, such as eosinophils, may be a good target to study the initiation of Th2 response. PMID- 19564894 TI - How do synapses measure milliseconds? PMID- 19564895 TI - Parks and tourism. PMID- 19564896 TI - A global perspective on trends in nature-based tourism. AB - Reports of rapid growth in nature-based tourism and recreation add significant weight to the economic case for biodiversity conservation but seem to contradict widely voiced concerns that people are becoming increasingly isolated from nature. This apparent paradox has been highlighted by a recent study showing that on a per capita basis, visits to natural areas in the United States and Japan have declined over the last two decades. These results have been cited as evidence of "a fundamental and pervasive shift away from nature-based recreation" but how widespread is this phenomenon? We address this question by looking at temporal trends in visitor numbers at 280 protected areas (PAs) from 20 countries. This more geographically representative dataset shows that while PA visitation (whether measured as total or per capita visit numbers) is indeed declining in the United States and Japan, it is generally increasing elsewhere. Total visit numbers are growing in 15 of the 20 countries for which we could get data, with the median national rate of change unrelated to the national rate of population growth but negatively associated with wealth. Reasons for this reversal of growth in the richest countries are difficult to pin down with existing data, but the pattern is mirrored by trends in international tourist arrivals as a whole and so may not necessarily be caused by disaffection with nature. Irrespective of the explanation, it is clear that despite important downturns in some countries, nature-related tourism is far from declining everywhere, and may still have considerable potential both to generate funds for conservation and to shape people's attitudes to the environment. PMID- 19564897 TI - RIN4 functions with plasma membrane H+-ATPases to regulate stomatal apertures during pathogen attack. AB - Pathogen perception by the plant innate immune system is of central importance to plant survival and productivity. The Arabidopsis protein RIN4 is a negative regulator of plant immunity. In order to identify additional proteins involved in RIN4-mediated immune signal transduction, we purified components of the RIN4 protein complex. We identified six novel proteins that had not previously been implicated in RIN4 signaling, including the plasma membrane (PM) H(+)-ATPases AHA1 and/or AHA2. RIN4 interacts with AHA1 and AHA2 both in vitro and in vivo. RIN4 overexpression and knockout lines exhibit differential PM H(+)-ATPase activity. PM H(+)-ATPase activation induces stomatal opening, enabling bacteria to gain entry into the plant leaf; inactivation induces stomatal closure thus restricting bacterial invasion. The rin4 knockout line exhibited reduced PM H(+) ATPase activity and, importantly, its stomata could not be re-opened by virulent Pseudomonas syringae. We also demonstrate that RIN4 is expressed in guard cells, highlighting the importance of this cell type in innate immunity. These results indicate that the Arabidopsis protein RIN4 functions with the PM H(+)-ATPase to regulate stomatal apertures, inhibiting the entry of bacterial pathogens into the plant leaf during infection. PMID- 19564898 TI - Can we "hedge" against the development of antiviral resistance among pandemic influenza viruses? PMID- 19564900 TI - Seasonal hunger: a neglected problem with proven solutions. PMID- 19564899 TI - Plant insecticide L-canavanine repels Drosophila via the insect orphan GPCR DmX. AB - For all animals, the taste sense is crucial to detect and avoid ingesting toxic molecules. Many toxins are synthesized by plants as a defense mechanism against insect predation. One example of such a natural toxic molecule is L-canavanine, a nonprotein amino acid found in the seeds of many legumes. Whether and how insects are informed that some plants contain L-canavanine remains to be elucidated. In insects, the taste sense relies on gustatory receptors forming the gustatory receptor (Gr) family. Gr proteins display highly divergent sequences, suggesting that they could cover the entire range of tastants. However, one cannot exclude the possibility of evolutionarily independent taste receptors. Here, we show that L-canavanine is not only toxic, but is also a repellent for Drosophila. Using a pharmacogenetic approach, we find that flies sense food containing this poison by the DmX receptor. DmXR is an insect orphan G-protein-coupled receptor that has partially diverged in its ligand binding pocket from the metabotropic glutamate receptor family. Blockade of DmXR function with an antagonist lowers the repulsive effect of L-canavanine. In addition, disruption of the DmXR encoding gene, called mangetout (mtt), suppresses the L-canavanine repellent effect. To avoid the ingestion of L-canavanine, DmXR expression is required in bitter sensitive gustatory receptor neurons, where it triggers the premature retraction of the proboscis, thus leading to the end of food searching. These findings show that the DmX receptor, which does not belong to the Gr family, fulfills a gustatory function necessary to avoid eating a natural toxin. PMID- 19564901 TI - The finer points of filopodia. PMID- 19564902 TI - Pralidoxime in acute organophosphorus insecticide poisoning--a randomised controlled trial. AB - BACKGROUND: Poisoning with organophosphorus (OP) insecticides is a major global public health problem, causing an estimated 200,000 deaths each year. Although the World Health Organization recommends use of pralidoxime, this antidote's effectiveness remains unclear. We aimed to determine whether the addition of pralidoxime chloride to atropine and supportive care offers benefit. METHODS AND FINDINGS: We performed a double-blind randomised placebo-controlled trial of pralidoxime chloride (2 g loading dose over 20 min, followed by a constant infusion of 0.5 g/h for up to 7 d) versus saline in patients with organophosphorus insecticide self-poisoning. Mortality was the primary outcome; secondary outcomes included intubation, duration of intubation, and time to death. We measured baseline markers of exposure and pharmacodynamic markers of response to aid interpretation of clinical outcomes. Two hundred thirty-five patients were randomised to receive pralidoxime (121) or saline placebo (114). Pralidoxime produced substantial and moderate red cell acetylcholinesterase reactivation in patients poisoned by diethyl and dimethyl compounds, respectively. Mortality was nonsignificantly higher in patients receiving pralidoxime: 30/121 (24.8%) receiving pralidoxime died, compared with 18/114 (15.8%) receiving placebo (adjusted hazard ratio [HR] 1.69, 95% confidence interval [CI] 0.88-3.26, p = 0.12). Incorporating the baseline amount of acetylcholinesterase already aged and plasma OP concentration into the analysis increased the HR for patients receiving pralidoxime compared to placebo, further decreasing the likelihood that pralidoxime is beneficial. The need for intubation was similar in both groups (pralidoxime 26/121 [21.5%], placebo 24/114 [21.1%], adjusted HR 1.27 [95% CI 0.71-2.29]). To reduce confounding due to ingestion of different insecticides, we further analysed patients with confirmed chlorpyrifos or dimethoate poisoning alone, finding no evidence of benefit. CONCLUSIONS: Despite clear reactivation of red cell acetylcholinesterase in diethyl organophosphorus pesticide poisoned patients, we found no evidence that this regimen improves survival or reduces need for intubation in patients with organophosphorus insecticide poisoning. The reason for this failure to benefit patients was not apparent. Further studies of different dose regimens or different oximes are required. PMID- 19564903 TI - Attention enhances the retrieval and stability of visuospatial and olfactory representations in the dorsal hippocampus. AB - A key question in the analysis of hippocampal memory relates to how attention modulates the encoding and long-term retrieval of spatial and nonspatial representations in this region. To address this question, we recorded from single cells over a period of 5 days in the CA1 region of the dorsal hippocampus while mice acquired one of two goal-oriented tasks. These tasks required the animals to find a hidden food reward by attending to either the visuospatial environment or a particular odor presented in shifting spatial locations. Attention to the visuospatial environment increased the stability of visuospatial representations and phase locking to gamma oscillations--a form of neuronal synchronization thought to underlie the attentional mechanism necessary for processing task relevant information. Attention to a spatially shifting olfactory cue compromised the stability of place fields and increased the stability of reward-associated odor representations, which were most consistently retrieved during periods of sniffing and digging when animals were restricted to the cup locations. Together, these results suggest that attention selectively modulates the encoding and retrieval of hippocampal representations by enhancing physiological responses to task-relevant information. PMID- 19564904 TI - Risk factors for helminth, malaria, and HIV infection in pregnancy in Entebbe, Uganda. AB - BACKGROUND: Infections during pregnancy may have serious consequences for both mother and baby. Assessment of risk factors for infections informs planning of interventions and analysis of the impact of infections on health outcomes. OBJECTIVES: To describe risk factors for helminths, malaria and HIV in pregnant Ugandan women before intervention in a trial of de-worming in pregnancy. METHODS: The trial recruited 2,507 pregnant women between April 2003 and November 2005. Participants were interviewed and blood and stool samples obtained; location of residence at enrolment was mapped. Demographic, socioeconomic, behavioral and other risk factors were modelled using logistic regression. RESULTS: There was a high prevalence of helminth, malaria and HIV infection, as previously reported. All helminths and malaria parasitemia were more common in younger women, and education was protective against every infection. Place of birth and/or tribe affected all helminths in a pattern consistent with the geographical distribution of helminth infections in Uganda. Four different geohelminths (hookworm, Trichuris, Ascaris and Trichostrongylus) showed a downwards trend in prevalence during the enrolment period. There was a negative association between hookworm and HIV, and between hookworm and low CD4 count among HIV-positive women. Locally, high prevalence of schistosomiasis and HIV occurred in lakeshore communities. CONCLUSIONS: Interventions for helminths, malaria and HIV need to target young women both in and out of school. Antenatal interventions for malaria and HIV infection must continue to be promoted. Women originating from a high risk area for a helminth infection remain at high risk after migration to a lower risk area, and vice versa, but overall, geohelminths seem to be becoming less common in this population. High risk populations, such as fishing communities, require directed effort against schistosomiasis and HIV infection. PMID- 19564906 TI - Tissue specific profiling of females of Schistosoma japonicum by integrated laser microdissection microscopy and microarray analysis. AB - BACKGROUND: The functions of many schistosome gene products remain to be characterized. A major step towards elucidating function of these genes would be in defining their sites of expression. This goal is rendered difficult to achieve by the generally small size of the parasites and the lack of a body cavity, which precludes analysis of transcriptional profiles of the tissues in isolation. METHODOLOGY/PRINCIPAL FINDINGS: Here, we describe a combined laser microdissection microscopy (LMM) and microarray analysis approach to expedite tissue specific profiling and gene atlasing for tissues of adult female Schistosoma japonicum. This approach helps to solve the gene characterization "bottle-neck" brought about by acoelomy and the size of these parasites. Complementary RNA obtained after isolation from gastrodermis (parasite gut mucosa), vitelline glands and ovary by LMM were subjected to microarray analyses, resulting in identification of 147 genes upregulated in the gastrodermis, 4,149 genes in the ovary and 2,553 in the vitellaria. CONCLUSIONS: This work will help to shed light on the molecular pathobiology of this debilitating human parasite and aid in the discovery of new targets for the development of anti-schistosome vaccines and drugs. PMID- 19564907 TI - Clean water should be recognized as a human right. PMID- 19564905 TI - Eps8 regulates axonal filopodia in hippocampal neurons in response to brain derived neurotrophic factor (BDNF). AB - The regulation of filopodia plays a crucial role during neuronal development and synaptogenesis. Axonal filopodia, which are known to originate presynaptic specializations, are regulated in response to neurotrophic factors. The structural components of filopodia are actin filaments, whose dynamics and organization are controlled by ensembles of actin-binding proteins. How neurotrophic factors regulate these latter proteins remains, however, poorly defined. Here, using a combination of mouse genetic, biochemical, and cell biological assays, we show that genetic removal of Eps8, an actin-binding and regulatory protein enriched in the growth cones and developing processes of neurons, significantly augments the number and density of vasodilator-stimulated phosphoprotein (VASP)-dependent axonal filopodia. The reintroduction of Eps8 wild type (WT), but not an Eps8 capping-defective mutant, into primary hippocampal neurons restored axonal filopodia to WT levels. We further show that the actin barbed-end capping activity of Eps8 is inhibited by brain-derived neurotrophic factor (BDNF) treatment through MAPK-dependent phosphorylation of Eps8 residues S624 and T628. Additionally, an Eps8 mutant, impaired in the MAPK target sites (S624A/T628A), displays increased association to actin-rich structures, is resistant to BDNF-mediated release from microfilaments, and inhibits BDNF-induced filopodia. The opposite is observed for a phosphomimetic Eps8 (S624E/T628E) mutant. Thus, collectively, our data identify Eps8 as a critical capping protein in the regulation of axonal filopodia and delineate a molecular pathway by which BDNF, through MAPK-dependent phosphorylation of Eps8, stimulates axonal filopodia formation, a process with crucial impacts on neuronal development and synapse formation. PMID- 19564908 TI - Venezuelan equine encephalitis in Panama: fatal endemic disease and genetic diversity of etiologic viral strains. AB - Venezuelan equine encephalitis (VEE) is a reemerging, mosquito-borne viral disease of the neotropics that is severely debilitating and sometimes fatal to humans. Periodic epidemics mediated by equine amplification have been recognized since the 1920s, but interepidemic disease is rarely recognized. We report here clinical findings and genetic characterization of 42 cases of endemic VEE detected in Panama from 1961-2004. Recent clusters of cases occurred in Darien (eastern Panama) and Panama provinces (central Panama) near rainforest and swamp habitats. Patients ranged from 10 months to 48 years of age, and the more severe cases with neurological complications, including one fatal infection, were observed in children. The VEE virus strains isolated from these cases all belonged to an enzootic, subtype ID lineage known to circulate among sylvatic vectors and rodent reservoir hosts in Panama and Peru. These findings underscore endemic VEE as an important but usually neglected arboviral disease of Latin America. PMID- 19564909 TI - The neovolcanic axis is a barrier to gene flow among Aedes aegypti populations in Mexico that differ in vector competence for Dengue 2 virus. AB - BACKGROUND: Aedes aegypti is the main mosquito vector of the four serotypes of dengue virus (DENV). Previous population genetic and vector competence studies have demonstrated substantial genetic structure and major differences in the ability to transmit dengue viruses in Ae. aegypti populations in Mexico. METHODOLOGY/PRINCIPAL FINDINGS: Population genetic studies revealed that the intersection of the Neovolcanic axis (NVA) with the Gulf of Mexico coast in the state of Veracruz acts as a discrete barrier to gene flow among Ae. aegypti populations north and south of the NVA. The mosquito populations north and south of the NVA also differed in their vector competence (VC) for dengue serotype 2 virus (DENV2). The average VC rate for Ae. aegypti mosquitoes from populations from north of the NVA was 0.55; in contrast the average VC rate for mosquitoes from populations from south of the NVA was 0.20. Most of this variation was attributable to a midgut infection and escape barriers. In Ae. aegypti north of the NVA 21.5% failed to develop midgut infections and 30.3% of those with an infected midgut failed to develop a disseminated infection. In contrast, south of the NVA 45.2% failed to develop midgut infections and 62.8% of those with an infected midgut failed to develop a disseminated infection. CONCLUSIONS: Barriers to gene flow in vector populations may also impact the frequency of genes that condition continuous and epidemiologically relevant traits such as vector competence. Further studies are warranted to determine why the NVA is a barrier to gene flow and to determine whether the differences in vector competence seen north and south of the NVA are stable and epidemiologically significant. PMID- 19564910 TI - The hedgehog signaling pathway: where did it come from? PMID- 19564911 TI - Molecular dynamics simulation of ligand dissociation from liver fatty acid binding protein. AB - The mechanisms of how ligands enter and leave the binding cavity of fatty acid binding proteins (FABPs) have been a puzzling question over decades. Liver fatty acid binding protein (LFABP) is a unique family member which accommodates two molecules of fatty acids in its cavity and exhibits the capability of interacting with a variety of ligands with different chemical structures and properties. Investigating the ligand dissociation processes of LFABP is thus a quite interesting topic, which however is rather difficult for both experimental approaches and ordinary simulation strategies. In the current study, random expulsion molecular dynamics simulation, which accelerates ligand motions for rapid dissociation, was used to explore the potential egress routes of ligands from LFABP. The results showed that the previously hypothesized "portal region" could be readily used for the dissociation of ligands at both the low affinity site and the high affinity site. Besides, one alternative portal was shown to be highly favorable for ligand egress from the high affinity site and be related to the unique structural feature of LFABP. This result lends strong support to the hypothesis from the previous NMR exchange studies, which in turn indicates an important role for this alternative portal. Another less favored potential portal located near the N-terminal end was also identified. Identification of the dissociation pathways will allow further mechanistic understanding of fatty acid uptake and release by computational and/or experimental techniques. PMID- 19564912 TI - Metabolites of an Epac-selective cAMP analog induce cortisol synthesis by adrenocortical cells through a cAMP-independent pathway. AB - Adrenal zona fasciculata (AZF) cells express a cAMP-activated guanine nucleotide exchange protein (Epac2) that may function in ACTH-stimulated cortisol synthesis. Experiments were done to determine whether cAMP analogs that selectively activate Epacs could induce cortisol synthesis and the expression of genes coding for steroidogenic proteins in bovine AZF cells. Treatment of AZF cells with the Epac selective cAMP analog (ESCA) 8CPT-2'-OMe-cAMP induced large (>100 fold), concentration-dependent, delayed increases in cortisol synthesis and the expression of mRNAs coding for the steroid hydroxylases CYP11a1, CYP17, CYP21, and the steroid acute regulatory protein (StAR). However, a non-hydrolyzable analog of this ESCA, Sp-8CPT-2'-OMe-cAMP, failed to stimulate cortisol production even at concentrations that activated Rap1, a downstream effector of Epac2. Accordingly, putative metabolites of 8CPT-2'-OMe-cAMP, including 8CPT-2'-OMe 5'AMP, 8CPT-2'-OMe-adenosine, and 8CPT-adenine all induced cortisol synthesis and steroid hydroxylase mRNA expression with a temporal pattern, potency, and effectiveness similar to the parent compound. At concentrations that markedly stimulated cortisol production, none of these metabolites significantly activated cAMP-dependent protein kinase (PKA). These results show that one or more metabolites of the ESCA 8CPT-2'-OMe-cAMP induce cortico-steroidogenesis by activating a panel of genes that code for steroidogenic proteins. The remarkable increases in cortisol synthesis observed in this study appear to be mediated by a novel cAMP-, Epac- and PKA-independent signaling pathway. PMID- 19564913 TI - Boule is present in fish and bisexually expressed in adult and embryonic germ cells of medaka. AB - BACKGROUND: The DAZ family genes boule, daz and dazl encode RNA binding proteins essential for fertility of diverse animals including human. dazl has bisexual expression in both mitotic and meiotic germ cells, whereas daz has male premeiotic expression, and boule is largely a unisexual meiotic regulator. Although boule has been proposed as the ancestor for dazl/daz by gene duplication, it has been identified only in invertebrates and mammals. It has, however, remained unclear when and how the DAZ family has evolved in vertebrates. METHODOLOGY AND PRINCIPAL FINDINGS: This study was aimed at identifying and characterizing the DAZ family genes in fish as the basal vertebrate. We show that boule and dazl coexist in medaka and stickleback. Similar to the medaka dazl (Odazl), the medaka boule (Obol) is maternally supplied and segregates with primordial germ cells. Surprisingly, Obol is expressed in adult germ cells at pre meiotic and meiotic stages of spermatogenesis and oogenesis. However, the maximal meiotic Obol expression in spermatocytes contrasts with the predominant pre meiotic Odazl expression in spermatogonia, and the diffuse cytoplasmic Obol distribution in early oocytes contrasts with the Odazl concentration in the Balbinani's body. CONCLUSIONS: The identification of fish boule and dazl genes provides direct evidence for the early gene duplication during vertebrate evolution. Our finding that Obol exhibits bisexual expression in both embryonic and adult germ cells considerably extends the diversity of boule expression patterns and offers a new insight into the evolutions of DAZ family members, expression patterns and functions in animal fertility. PMID- 19564914 TI - Transcription-independent heritability of induced histone modifications in the mouse preimplantation embryo. AB - Enzyme-catalyzed, post-translational modifications of core histones have been implicated in the complex changes in gene expression that drive early mammalian development. However, until recently the small number of cells available from the preimplantation embryo itself has prevented quantitative analysis of histone modifications at key regulator genes. The possible involvement of histone modifications in the embryo's response to extracellular signals, or as determinants of cell fate or lineage progression, remains unclear. Here we describe the use of a recently-developed chromatin immunoprecipitation technique (CChIP) to assay histone modification levels at key regulator genes (Pou5f1, Nanog, Cdx2, Hoxb1, Hoxb9) as mouse embryos progress from 8-cell to blastocyst in culture. Only by the blastocyst stage, when the embryonic (Inner Cell Mass) and extra-embryonic (Trophoblast) lineages are compared, do we see the expected association between histone modifications previously linked to active and silent chromatin, and transcriptional state. To explore responses to an environmental signal, we exposed embryos to the histone deacetylase inhibitor, anti-epileptic and known teratogen valproic acid (VPA), during progression from 8-cell to morula stage. Such treatment increased H4 acetylation and H3 lysine 4 methylation at the promoters of Hoxb1 and Hoxb9, but not the promoters of Pou5f1, Nanog,Cdx2 or the housekeeping gene Gapdh. Despite the absence of detectable Hoxb transcription, these VPA-induced changes were heritable, following removal of the inhibitor, at least until the blastocyst stage. The selective hyperacetylation of Hoxb promoters in response to a histone deacetylase inhibitor, suggests that Hox genes have a higher turnover of histone acetates than other genes in the preimplantation embryo. To explain the heritability, through mitosis, of VPA induced changes in histone modification at Hoxb promoters, we describe how an epigenetic feed-forward loop, based on cross-talk between H3 acetylation and H3K4 methylation, might generate a persistently increased steady-state level of histone acetylation in response to a transient signal. PMID- 19564915 TI - Racial differences in the human endogenous circadian period. AB - The length of the endogenous period of the human circadian clock (tau) is slightly greater than 24 hours. There are individual differences in tau, which influence the phase angle of entrainment to the light/dark (LD) cycle, and in doing so contribute to morningness-eveningness. We have recently reported that tau measured in subjects living on an ultradian LD cycle averaged 24.2 hours, and is similar to tau measured using different experimental methods. Here we report racial differences in tau. Subjects lived on an ultradian LD cycle (1.5 hours sleep, 2.5 hours wake) for 3 days. Circadian phase assessments were conducted before and after the ultradian days to determine the change in circadian phase, which was attributed to tau. African American subjects had a significantly shorter tau than subjects of other races. We also tested for racial differences in our previous circadian phase advancing and phase delaying studies. In the phase advancing study, subjects underwent 4 days of a gradually advancing sleep schedule combined with a bright light pulse upon awakening each morning. In the phase delaying study, subjects underwent 4 days of a gradually delaying sleep schedule combined with evening light pulses before bedtime. African American subjects had larger phase advances and smaller phase delays, relative to Caucasian subjects. The racial differences in tau and circadian phase shifting have important implications for understanding normal phase differences between individuals, for developing solutions to the problems of jet lag and shift work, and for the diagnosis and treatment of circadian rhythm based sleep disorders such as advanced and delayed sleep phase disorder. PMID- 19564916 TI - A MAP kinase dependent feedback mechanism controls Rho1 GTPase and actin distribution in yeast. AB - In the yeast Saccharomyces cerevisiae the guanosine triphosphatase (GTPase) Rho1 controls actin polarization and cell wall expansion. When cells are exposed to various environmental stresses that perturb the cell wall, Rho1 activates Pkc1, a mammalian Protein Kinase C homologue, and Mpk1, a mitogen activated protein kinase (MAPK), resulting in actin depolarization and cell wall remodeling. In this study, we demonstrate a novel feedback loop in this Rho1-mediated Pkc1-MAPK pathway that involves regulation of Rom2, the guanine nucleotide exchange factor of Rho1, by Mpk1, the end kinase of the pathway. This previously unrecognized Mpk1-dependent feedback is a critical step in regulating Rho1 function. Activation of this feedback mechanism is responsible for redistribution of Rom2 and cell wall synthesis activity from the bud to cell periphery under stress conditions. It is also required for terminating Rho1 activity toward the Pkc1 MAPK pathway and for repolarizing actin cytoskeleton and restoring growth after the stressed cells become adapted. PMID- 19564917 TI - Hemichannel-mediated and pH-based feedback from horizontal cells to cones in the vertebrate retina. AB - BACKGROUND: Recent studies designed to identify the mechanism by which retinal horizontal cells communicate with cones have implicated two processes. According to one account, horizontal cell hyperpolarization induces an increase in pH within the synaptic cleft that activates the calcium current (Ca(2+)-current) in cones, enhancing transmitter release. An alternative account suggests that horizontal cell hyperpolarization increases the Ca(2+)-current to promote transmitter release through a hemichannel-mediated ephaptic mechanism. METHODOLOGY/PRINCIPAL FINDINGS: To distinguish between these mechanisms, we interfered with the pH regulating systems in the retina and studied the effects on the feedback responses of cones and horizontal cells. We found that the pH buffers HEPES and Tris partially inhibit feedback responses in cones and horizontal cells and lead to intracellular acidification of neurons. Application of 25 mM acetate, which does not change the extracellular pH buffer capacity, does lead to both intracellular acidification and inhibition of feedback. Because intracellular acidification is known to inhibit hemichannels, the key experiment used to test the pH hypothesis, i.e. increasing the extracellular pH buffer capacity, does not discriminate between a pH-based feedback system and a hemichannel-mediated feedback system. To test the pH hypothesis in a manner independent of artificial pH-buffer systems, we studied the effect of interfering with the endogenous pH buffer, the bicarbonate/carbonic anhydrase system. Inhibition of carbonic anhydrase allowed for large changes in pH in the synaptic cleft of bipolar cell terminals and cone terminals, but the predicted enhancement of the cone feedback responses, according to the pH-hypothesis, was not observed. These experiments thus failed to support a proton mediated feedback mechanism. The alternative hypothesis, the hemichannel-mediated ephaptic feedback mechanism, was therefore studied experimentally, and its feasibility was buttressed by means of a quantitative computer model of the cone/horizontal cell synapse. CONCLUSION: We conclude that the data presented in this paper offers further support for physiologically relevant ephaptic interactions in the retina. PMID- 19564918 TI - Dopamine transporters in striatum correlate with deactivation in the default mode network during visuospatial attention. AB - BACKGROUND: Dopamine and dopamine transporters (DAT, which regulate extracellular dopamine in the brain) are implicated in the modulation of attention but their specific roles are not well understood. Here we hypothesized that dopamine modulates attention by facilitation of brain deactivation in the default mode network (DMN). Thus, higher striatal DAT levels, which would result in an enhanced clearance of dopamine and hence weaker dopamine signals, would be associated to lower deactivation in the DMN during an attention task. METHODOLOGY/PRINCIPAL FINDINGS: For this purpose we assessed the relationship between DAT in striatum (measured with positron emission tomography and [(11)C]cocaine used as DAT radiotracer) and brain activation and deactivation during a parametric visual attention task (measured with blood oxygenation level dependent functional magnetic resonance imaging) in healthy controls. We show that DAT availability in caudate and putamen had a negative correlation with deactivation in ventral parietal regions of the DMN (precuneus, BA 7) and a positive correlation with deactivation in a small region in the ventral anterior cingulate gyrus (BA 24/32). With increasing attentional load, DAT in caudate showed a negative correlation with load-related deactivation increases in precuneus. CONCLUSIONS/SIGNIFICANCE: These findings provide evidence that dopamine transporters modulate neural activity in the DMN and anterior cingulate gyrus during visuospatial attention. Our findings suggest that dopamine modulates attention in part by regulating neuronal activity in posterior parietal cortex including precuneus (region involved in alertness) and cingulate gyrus (region deactivated in proportion to emotional interference). These findings suggest that the beneficial effects of stimulant medications (increase dopamine by blocking DAT) in inattention reflect in part their ability to facilitate the deactivation of the DMN. PMID- 19564919 TI - Methamphetamine induces dopamine D1 receptor-dependent endoplasmic reticulum stress-related molecular events in the rat striatum. AB - Methamphetamine (METH) is an illicit toxic psychostimulant which is widely abused. Its toxic effects depend on the release of excessive levels of dopamine (DA) that activates striatal DA receptors. Inhibition of DA-mediated neurotransmission by the DA D1 receptor antagonist, SCH23390, protects against METH-induced neuronal apoptosis. The initial purpose of the present study was to investigate, using microarray analyses, the influence of SCH23390 on transcriptional responses in the rat striatum caused by a single METH injection at 2 and 4 hours after drug administration. We identified 545 out of a total of 22,227 genes as METH-responsive. These include genes which are involved in apoptotic pathways, endoplasmic reticulum (ER) stress, and in transcription regulation, among others. Of these, a total of 172 genes showed SCH23390-induced inhibition of METH-mediated changes. Among these SCH23390-responsive genes were several genes that are regulated during ER stress, namely ATF3, HSP27, Hmox1, HSP40, and CHOP/Gadd153. The secondary goal of the study was to investigate the role of DA D1 receptor stimulation on the expression of genes that participate in ER stress-mediated molecular events. We thus used quantitative PCR to confirm changes in the METH-responsive ER genes identified by the microarray analyses. We also measured the expression of these genes and of ATF4, ATF6, BiP/GRP78, and of GADD34 over a more extended time course. SCH23390 attenuated or blocked METH induced increases in the expression of the majority of these genes. Western blot analysis revealed METH-induced increases in the expression of the antioxidant protein, Hmox1, which lasted for about 24 hours after the METH injection. Additionally, METH caused DA D1 receptor-dependent transit of the Hmox1 regulator protein, Nrf2, from cytosolic into nuclear fractions where the protein exerts its regulatory functions. When taken together, these findings indicate that SCH23390 can provide protection against neuronal apoptosis by inhibiting METH-mediated DA D1 receptor-mediated ER stress in the rat striatum. Our data also suggest that METH-induced toxicity might be a useful model to dissect molecular mechanisms involved in ER stress-dependent events in the rodent brain. PMID- 19564920 TI - Bacterial ortholog of mammalian translocator protein (TSPO) with virulence regulating activity. AB - The translocator protein (TSPO), previously designated as peripheral-type benzodiazepine receptor, is a protein mainly located in the outer mitochondrial membrane of eukaryotic cells. TSPO is implicated in major physiological functions and functionally associated with other proteins such as the voltage-dependent anionic channel, also designated as mitochondrial porin. Surprisingly, a TSPO related protein was identified in the photosynthetic bacterium Rhodobacter sphaeroides but it was initially considered as a relict of evolution. In the present study we cloned a tspO gene in Pseudomonas fluorescens MF37, a non photosynthetic eubacterium and we used bioinformatics tools to identify TSPO in the genome of 97 other bacteria. P. fluorescens TSPO was recognized by antibodies against mouse protein and by PK 11195, an artificial ligand of mitochondrial TSPO. As in eukaryotes, bacterial TSPO appears functionally organized as a dimer and the apparent Kd for PK 11195 is in the same range than for its eukaryotic counterpart. When P. fluorescens MF37 was treated with PK 11195 (10(-5) M) adhesion to living or artificial surfaces and biofilm formation activity were increased. Conversely, the apoptotic potential of bacteria on eukaryotic cells was significantly reduced. This effect of PK11195 was abolished in a mutant of P. fluorescens MF37 deficient for its major outer membrane porin, OprF. The present results demonstrate the existence of a bacterial TSPO that shares common structural and functional characteristics with its mammalian counterpart. This protein, apparently involved in adhesion and virulence, reveals the existence of a possible new inter kingdom signalling system and suggests that the human microbiome should be involuntarily exposed to the evolutionary pressure of benzodiazepines and related molecules. This discovery also represents a promising opportunity for the development of alternative antibacterial strategies. PMID- 19564921 TI - Generation of monoclonal antibodies against highly conserved antigens. AB - BACKGROUND: Therapeutic antibody development is one of the fastest growing areas of the pharmaceutical industry. Generating high-quality monoclonal antibodies against a given therapeutic target is very crucial for the success of the drug development. However, due to immune tolerance, some proteins that are highly conserved between mice and humans are not very immunogenic in mice, making it difficult to generate antibodies using a conventional approach. METHODOLOGY/PRINCIPAL FINDINGS: In this report, the impaired immune tolerance of NZB/W mice was exploited to generate monoclonal antibodies against highly conserved or self-antigens. Using two highly conserved human antigens (MIF and HMGB1) and one mouse self-antigen (TNF-alpha) as examples, we demonstrate here that multiple clones of high affinity, highly specific antibodies with desired biological activities can be generated, using the NZB/W mouse as the immunization host and a T cell-specific tag fused to a recombinant antigen to stimulate the immune system. CONCLUSIONS/SIGNIFICANCE: We developed an efficient and universal method for generating surrogate or therapeutic antibodies against "difficult antigens" to facilitate the development of therapeutic antibodies. PMID- 19564923 TI - Perception of neighborhood safety and reported childhood lifetime asthma in the United States (U.S.): a study based on a national survey. AB - BACKGROUND: Recent studies have emphasized the role of psychosocial stressors as a determinant of asthma, and neighborhoods can be a potential source of such stressors. We investigated the association between parental perception of neighborhood safety and reported lifetime asthma among children. METHODOLOGY/PRINCIPAL FINDINGS: Data for the study came from the 2003-04 National Survey of Children Health (NSCH); a nationally representative cross-sectional sample of children aged 0-17 years. Demographic, socioeconomic and behavioral covariates were included in the study. Models were estimated after taking account of weighting and complex survey design. Parental report of whether the child has ever been diagnosed with asthma by a physician was used to define the outcome. Parental report of perception of neighborhood safety was the main exposure. In unadjusted models, the odds ratio (OR) for reporting asthma associated with living in neighborhoods that were perceived to be sometimes or never safe was 1.36 (95% confidence intervals [CI] 1.21, 1.53) compared to living in neighborhoods that were perceived to be always safe. Adjusting for covariates including exposure to second hand tobacco smoke, mother's self-rated health, child's physical activity and television viewing attenuated this association (OR 1.25, 95% CI 1.08, 1.43). In adjusted models, the increased odds ratio for reporting asthma was also higher among those who perceived neighborhoods as being usually safe (OR 1.15 95% CI 1.06, 1.26), as compared to always safe, suggestive of a dose-response relationship, with the differentials for usually safe and never safe being statistically significant (p = 0.009). CONCLUSION: Psychosocial stressors may be important risk factors that may impact the pathogenesis of asthma and/or contribute to asthma morbidity by triggering exacerbations through neuroimmunologic mechanisms, as well as social mechanisms. PMID- 19564924 TI - Early cell fate decisions of human embryonic stem cells and mouse epiblast stem cells are controlled by the same signalling pathways. AB - Human embryonic stem cells have unique value for regenerative medicine, as they are capable of differentiating into a broad variety of cell types. Therefore, defining the signalling pathways that control early cell fate decisions of pluripotent stem cells represents a major task. Moreover, modelling the early steps of embryonic development in vitro may provide the best approach to produce cell types with native properties. Here, we analysed the function of key developmental growth factors such as Activin, FGF and BMP in the control of early cell fate decisions of human pluripotent stem cells. This analysis resulted in the development and validation of chemically defined culture conditions for achieving specification of human embryonic stem cells into neuroectoderm, mesendoderm and into extra-embryonic tissues. Importantly, these defined culture conditions are devoid of factors that could obscure analysis of developmental mechanisms or render the resulting tissues incompatible with future clinical applications. Importantly, the growth factor roles defined using these culture conditions similarly drove differentiation of mouse epiblast stem cells derived from post implantation embryos, thereby reinforcing the hypothesis that epiblast stem cells share a common embryonic identity with human pluripotent stem cells. Therefore the defined growth factor conditions described here represent an essential step toward the production of mature cell types from pluripotent stem cells in conditions fully compatible with clinical use ant also provide a general approach for modelling the early steps of mammalian embryonic development. PMID- 19564922 TI - Synergistic activation of HIV-1 expression by deacetylase inhibitors and prostratin: implications for treatment of latent infection. AB - The persistence of transcriptionally silent but replication-competent HIV-1 reservoirs in Highly Active Anti-Retroviral Therapy (HAART)-treated infected individuals, represents a major hurdle to virus eradication. Activation of HIV-1 gene expression in these cells together with an efficient HAART has been proposed as an adjuvant therapy aimed at decreasing the pool of latent viral reservoirs. Using the latently-infected U1 monocytic cell line and latently-infected J-Lat T cell clones, we here demonstrated a strong synergistic activation of HIV-1 production by clinically used histone deacetylase inhibitors (HDACIs) combined with prostratin, a non-tumor-promoting nuclear factor (NF)- kappaB inducer. In J Lat cells, we showed that this synergism was due, at least partially, to the synergistic recruitment of unresponsive cells into the expressing cell population. A combination of prostratin+HDACI synergistically activated the 5' Long Terminal Repeat (5'LTR) from HIV-1 Major group subtypes representing the most prevalent viral genetic forms, as shown by transient transfection reporter assays. Mechanistically, HDACIs increased prostratin-induced DNA-binding activity of nuclear NF-kappaB and degradation of cytoplasmic NF-kappaB inhibitor, IkappaBalpha . Moreover, the combined treatment prostratin+HDACI caused a more pronounced nucleosomal remodeling in the U1 viral promoter region than the treatments with the compounds alone. This more pronounced remodeling correlated with a synergistic reactivation of HIV-1 transcription following the combined treatment prostratin+HDACI, as demonstrated by measuring recruitment of RNA polymerase II to the 5'LTR and both initiated and elongated transcripts. The physiological relevance of the prostratin+HDACI synergism was shown in CD8(+) depleted peripheral blood mononuclear cells from HAART-treated patients with undetectable viral load. Moreover, this combined treatment reactivated viral replication in resting CD4(+) T cells isolated from similar patients. Our results suggest that combinations of different kinds of proviral activators may have important implications for reducing the size of latent HIV-1 reservoirs in HAART treated patients. PMID- 19564925 TI - HCV antibody response and genotype distribution in different areas and races of China. AB - Hepatitis C virus (HCV) heterogeneity accounts for the failure of effective vaccine development and the lack of successful anti-viral therapy in some patients. Little is known about the immune response to HCV peptides and the region or race specific genotypes in China. The objective of this study was to characterize HCV antibody immune response to HCV peptides and HCV genotypes in different regions and races of China. A total of 363 serum samples were collected from HCV carriers in 6 regions in China. The immune response to HCV peptides was evaluated by ELISA. HCV genotypes were examined using nested RT-PCR. We found that the anti-HCV antibody neutralization rates were significantly different among the serum samples from different areas or from different races in the same area. For samples from Tibet and Sinkiang, the rates of neutralization by HCV peptides were only 3.2% and 30.8%, respectively. The genotypes of samples from Tibet and Sinkiang were apparently heterogeneic and included type I, II, III and multiple types (I/II/III, I/II, I/III, II/III). One specific sample with multiple genotype (I/II/III) HCV infection was found to consist of type I, II, III, II/III and an unclassified genotype. These studies indicate that the anti-HCV antibody immune response to HCV peptides varied across regions and among races. The distribution of HCV genotypes among Tibetans in Tibet and Uighurs in Sinkiang was different from that in the inner areas of China. In addition, a "master" genotype, type II, was found to exist in HCV infection with multiple HCV genotypes. PMID- 19564926 TI - Nuclear localization of p38 MAPK in response to DNA damage. AB - p38 MAP kinase (MAPK) is activated in response to environmental stress, cytokines and DNA damage, and mediates death, cell differentiation and cell cycle checkpoints. The intracellular localization of p38 MAPK upon activation remains unclear, and may depend on the stimulus. We show here that activation of p38 MAPK by stimuli that induce DNA double strand breaks (DSBs), but not other stimuli, leads to its nuclear translocation. In addition, naturally occurring DSBs generated through V(D)J recombination in immature thymocytes also promote nuclear accumulation of p38 MAPK. Nuclear translocation of p38 MAPK does not require its catalytic activity, but is induced by a conformational change of p38 MAPK triggered by phosphorylation within the active site. The selective nuclear accumulation of p38 MAPK in response to DNA damage could be a mechanism to facilitate the phosphorylation of p38 MAPK nuclear targets for the induction of a G2/M cell cycle checkpoint and DNA repair. PMID- 19564927 TI - Essential roles of mTOR/Akt pathway in Aurora-A cell transformation. AB - We have recently demonstrated that Aurora-A kinase is a potential oncogene to develop mammary gland tumors in mice, when expressed under MMTV promoter. These tumors contain phosphorylated forms of Akt and mTOR, suggesting that Akt-mTOR pathway is involved in transformed phenotype induced by Aurora-A. In the present studies, we discovered that stable cell lines expressing Aurora-A contain phosphorylation of Akt Ser473 after prolonged passages of cell culture, not in cells of the early period of cell culture. Levels of PTEN tumor suppressor are significantly reduced in these late passage cells at least in part due to increased poly ubiquitination of the protein. Akt-activated Aurora-A cells formed larger colonies in soft agar and are resistant to UV-induced apoptosis. Aurora-A inhibitor, VX-680, can cause cell death of Aurora-A cells in which Akt is not activated. siRNA-mediated depletion of mTOR in those cells resulted in decreased phosphorylation of Akt Ser473, suggesting that TORC2 complex phosphorylates Akt in Aurora-A cells. Treatment of late-passage Aurora-A cells with mTOR inhibitor reduced colony formation in soft agar. These results strongly suggest that commitment of cell transformation by Aurora-A is determined by at least co activation of Akt/mTOR pathway. PMID- 19564929 TI - Site-dependent differences in clinical, pathohistological, and molecular parameters in metastatic colon cancer. AB - The purpose was to develop a metastatic score specific to the hepatic and peritoneal site in colorectal cancer patients from clinical, pathohistological and molecular markers potentially reflecting oncogenic activation (OA) or epithelial-mesenchymal transition (EMT), where OA may reflect an activation and EMT the functional loss of certain genes. The primary tumour stage (OA, EMT), lymphonodal stage (OA), the presence of a lymphangiosis carcinomatosa (OA), histological grade (OA, EMT), and immunoblot extraction of E-cadherin (OA, EMT) were differentially rated with zero to one or two points due to their potential contribution to each process and the resulting scores were validated in 27 colorectal cancer patients (three patients with pre-malignant adenomas, 16 with primaries and two with local recurrencies, three of which were metastatic to the peritoneum, six metastatic to the liver and two metastatic to both, the liver and the peritoneum, and five with hepatic secondaries, one of which at histology was metastatic to the peritoneum too). As a single parameter only the N-stage significantly contributed to OA (p<0.05). Median OA and EMT scores, however, were 3.5 and 2 in the case of primaries without further spread, 5 and 4 in those nodal positive, 5 and 4 in the case of peritoneal implants, 6 and 2 in the case of liver metastases, and 6.5 and 3 in the case of a simultaneous hepatic and peritoneal spread, respectively. These differences were significant when scores from patients with and without liver metastases (OA, p<0.002) or with peritoneal implants and isolated hepatic spread (EMT, p<0.01) were compared. The results suggest a site-specific contribution of OA and EMT to tumour progression in human colon cancer. PMID- 19564930 TI - Glycosylation of fluorophenols by plant cell cultures. AB - Fluoroaromatic compounds are used as agrochemicals and released into environment as pollutants. Glycosylation of 2-, 3-, and 4-fluorophenols using plant cell cultures of Nicotiana tabacum was investigated to elucidate their potential to metabolize these compounds. Cultured N. tabacum cells converted 2-fluorophenol into its beta-glucoside (60%) and beta-gentiobioside (10%). 4-Fluorophenol was also glycosylated to its beta-glucoside (32%) and beta-gentiobioside (6%) by N. tabacum cells. On the other hand, N. tabacum glycosylated 3-fluorophenol to beta glucoside (17%). PMID- 19564928 TI - Construction of a full-length cDNA Library from Chinese oak silkworm pupa and identification of a KK-42-binding protein gene in relation to pupa-diapause termination. AB - In this study we successfully constructed a full-length cDNA library from Chinese oak silkworm, Antheraea pernyi, the most well-known wild silkworm used for silk production and insect food. Total RNA was extracted from a single fresh female pupa at the diapause stage. The titer of the library was 5 x 10(5) cfu/ml and the proportion of recombinant clones was approximately 95%. Expressed sequence tag (EST) analysis was used to characterize the library. A total of 175 clustered ESTs consisting of 24 contigs and 151 singlets were generated from 250 effective sequences. Of the 175 unigenes, 97 (55.4%) were known genes but only five from A. pernyi, 37 (21.2%) were known ESTs without function annotation, and 41 (23.4%) were novel ESTs. By EST sequencing, a gene coding KK-42-binding protein in A. pernyi (named as ApKK42-BP; GenBank accession no. FJ744151) was identified and characterized. Protein sequence analysis showed that ApKK42-BP was not a membrane protein but an extracellular protein with a signal peptide at position 1-18, and contained two putative conserved domains, abhydro_lipase and abhydrolase_1, suggesting it may be a member of lipase superfamily. Expression analysis based on number of ESTs showed that ApKK42-BP was an abundant gene in the period of diapause stage, suggesting it may also be involved in pupa-diapause termination. PMID- 19564931 TI - Self-assembly of optical molecules with supramolecular concepts. AB - Fabrication of nano-sized objects is one of the most important issues in nanoscience and nanotechnology. Soft nanomaterials with flexible properties have been given much attention and can be obtained through bottom-up processing from functional molecules, where self-assembly based on supramolecular chemistry and designed assembly have become crucial processes and techniques. Among the various functional molecules, dyes have become important materials in certain areas of nanotechnology and their self-assembling behaviors have been actively researched. In this short review, we briefly introduce recent progress in self-assembly of optical molecules and dyes, based mainly on supramolecular concepts. The introduced examples are classified into four categories: self-assembly of (i) low molecular-weight dyes and (ii) polymeric dyes and dye self-assembly (iii) in nanoscale architectures and (iv) at surfaces. PMID- 19564932 TI - Time-course expression profiles of hair cycle-associated genes in male mini rats after depilation of telogen-phase hairs. AB - Jcl:WistarTGN(ARGHGEN)1Nts rat (Mini rat) is a growth hormone (GH)-deficient transgenic rat. The hair cycle in the dorsal skin of male Mini rats enters a long lasting telogen phase after eights weeks of age, but depilation can induce a transient hair cycle again. In this study, a time-course profiling of genes expression was done on the dorsal skin of male Mini rats along the progression of depilation-induced hair cycle using DNA microarray analysis. As a result, 1,215 probe sets including 1,171 hair cycle-related ones showed more than 3-fold changes in expression compared with that in before-depilation telogen phase. The present data will contribute to elucidating the mechanisms of hair cycle regulation and should lead to the identification of novel molecular targets for hair growth and/or depilation agents. PMID- 19564933 TI - Current mathematical methods used in QSAR/QSPR studies. AB - This paper gives an overview of the mathematical methods currently used in quantitative structure-activity/property relationship (QASR/QSPR) studies. Recently, the mathematical methods applied to the regression of QASR/QSPR models are developing very fast, and new methods, such as Gene Expression Programming (GEP), Project Pursuit Regression (PPR) and Local Lazy Regression (LLR) have appeared on the QASR/QSPR stage. At the same time, the earlier methods, including Multiple Linear Regression (MLR), Partial Least Squares (PLS), Neural Networks (NN), Support Vector Machine (SVM) and so on, are being upgraded to improve their performance in QASR/QSPR studies. These new and upgraded methods and algorithms are described in detail, and their advantages and disadvantages are evaluated and discussed, to show their application potential in QASR/QSPR studies in the future. PMID- 19564934 TI - Localization of D-beta-aspartyl residue-containing proteins in various tissues. AB - Prior to the emergence of life, it is believed that only l-amino acids were selected for formation of protein and that d-amino acids were eliminated on the primitive Earth. Whilst homochirality is essential for life, the occurrence of proteins containing d-beta-aspartyl (Asp) residues in various tissues from elderly subjects has been reported recently. Here, we demonstrate the presence of a d-beta-Asp-containing protein in the cardiac muscle of heart, blood vessels of the lung, chief cells of the stomach, longitudinal and circular muscle of the stomach, small intestine and large intestine. Since the d-beta-Asp residue occurs through a succinimide intermediate, this isomer may potentially be generated in proteins more easily than initially thought. Formation of the d-beta-Asp residue in proteins may be related to stress. PMID- 19564935 TI - Soluble tumor necrosis factor receptor mediates cell proliferation on lipopolysaccharide-stimulated cultured human decidual stromal cells. AB - The tumor necrosis factor-alpha (TNF-alpha) cytokine receptor system modulates apoptosis in many cell types, so we have investigated the role of sTNFR1 in bacterial lipopolysaccharide (LPS)-induced cell death in cultured human decidual stromal cells, hypothesizing that sTNFR1 might play a central role in this action. In this work we characterized in vitro decidual stromal cell viability with LPS treatment and LPS and sTNFR1 co-treatment. We found that LPS treatment induced decidual stromal cell death in a dose-dependent manner and that sTNFR1 blocked the effect of the LPS treatment. There was a significant proliferation among cells co-incubated with LPS at 10 microg/mL and sTNFR1 at 0.1 microg/mL compared with LPS and sTNFR1 at 0.01, 0.05, 0.2 and 0.5 microg/mL (p < 0.01). This study demonstrated that LPS led to decidual stromal cell death in vitro but sTNFR1 down-regulates the cell death due to LPS under the same conditions. Taken together, these results suggested that sTNFR1 could participate in a protective mechanism against endotoxin. PMID- 19564936 TI - Concept of sustained ordering and an ATP-related mechanism of life's origin. AB - This paper shows that the steady state of a system of conjugated reactions, which are characterized by disproportionation of entropy and proceed in the domain of linear interactions, is an attractor of ordering. Such systems are primed to produce ordering, and life is a specific manifestation of the sustained ordering inherent to the chemistry of carbon. The adenosine triphospate (ATP) molecule has properties which makes ATP hydrolysis to be most appropriate to form such a system in primitive world. Hence, ATP is suggested to play a key role in prebiological evolution. Principles of the origin and evolution of life following from the concept of ordering are stated. PMID- 19564937 TI - Effects of hydroxylpropyl-beta-cyclodextrin on in vitro insulin stability. AB - The objective of this study was to elucidate the effects of hydroxylpropyl-beta cyclodextrin (HP-beta-CD) on the in vitro stability of insulin. It was found that HP-beta-CD had positive effects on the stability of insulin in acid and base and under high temperature conditions. Furthermore, use of HP-beta-CD could also increase the stability of disulfide bonds which are important to the conformation of insulin. Through (1)H-NMR experiments it was found that the protective effect of HP-beta-CD was due to complexation with insulin. The results suggest that the presence of HP-beta-CD could improve the stability of insulin in different environments. PMID- 19564938 TI - The mitochondrial protein translocation motor: structural conservation between the human and yeast Tim14/Pam18-Tim16/Pam16 co-chaperones. AB - Most of our knowledge regarding the process of protein import into mitochondria has come from research employing Saccharomyces cerevisiae as a model system. Recently, several mammalian homologues of the mitochondrial motor proteins were identified. Of particular interest for us is the human Tim14/Pam18-Tim16/Pam16 complex. We chose a structural approach in order to examine the evolutionary conservation between yeast Tim14/Pam18-Tim16/Pam16 proteins and their human homologues. For this purpose, we examined the structural properties of the purified human proteins and their interaction with their yeast homologues, in vitro. Our results show that the soluble domains of the human Tim14/Pam18 and Tim16/Pam16 proteins interact with their yeast counterparts, forming heterodimeric complexes and that these complexes interact with yeast mtHsp70. PMID- 19564939 TI - Rolly protein (ROLP)-Epb4.1/3: a potential protein-protein interaction relevant for the maintenance of cell adhesion. AB - We recently described Rolly Protein (ROLP), a small protein synthesized by substrate-adherent cells in a broad range of tissues. In a first set of experiments performed taking advantage of bone forming tibial cartilage as an experimental model we showed that ROLP transcription is associated to cells in an active proliferation state, whereas its downregulation is observed when cell proliferation decreases. Taking advantage of siRNA technology we also documented the expression modulation of some apoptosis-related genes in ROLP-silenced cells. In this work we search for the possible molecular interactors of ROLP by using both the antibody array approach as well as the co-immunoprecipitation approach. Results suggest the occurrence of an interaction of ROLP with Erythrocyte membrane Protein Band 4.1/3 (Epb4.1/3), an oncosuppressor downregulated in tumor development and in metastatic tissues; in addition we report experimental results that keep in line also with a potential interaction of ROLP with other PDZ containing proteins. We also present experimental evidences supporting a role played by ROLP in cell adhesion thus supporting the existence of a biologically relevant link between ROLP and Epb4.1/3. We here suggest that ROLP might exert its biological role cooperating with Epb4.1/3, a protein that is involved in biological pathways that are often inhibited in tumor metastasis. Given the role of Epb4.1/3 in contrasting cancerogenesis we think that its cooperation with ROLP might be relevant in cancer studies and deserves further investigation. PMID- 19564940 TI - GroEL-assisted protein folding: does it occur within the chaperonin inner cavity? AB - The folding of protein molecules in the GroEL inner cavity under the co chaperonin GroES lid is widely accepted as a crucial event of GroEL-assisted protein folding. This review is focused on the data showing that GroEL-assisted protein folding may proceed out of the complex with the chaperonin. The models of GroEL-assisted protein folding assuming ligand-controlled dissociation of nonnative proteins from the GroEL surface and their folding in the bulk solution are also discussed. PMID- 19564942 TI - Predicting complexation thermodynamic parameters of beta-cyclodextrin with chiral guests by using swarm intelligence and support vector machines. AB - The Particle Swarm Optimization (PSO) and Support Vector Machines (SVMs) approaches are used for predicting the thermodynamic parameters for the 1:1 inclusion complexation of chiral guests with beta-cyclodextrin. A PSO is adopted for descriptor selection in the quantitative structure-property relationships (QSPR) of a dataset of 74 chiral guests due to its simplicity, speed, and consistency. The modified PSO is then combined with SVMs for its good approximating properties, to generate a QSPR model with the selected features. Linear, polynomial, and Gaussian radial basis functions are used as kernels in SVMs. All models have demonstrated an impressive performance with R(2) higher than 0.8. PMID- 19564941 TI - A possible mechanism for evading temperature quantum decoherence in living matter by feshbach resonance. AB - A new possible scenario for the origin of the molecular collective behaviour associated with the emergence of living matter is presented. We propose that the transition from a non-living to a living cell could be mapped to a quantum transition to a coherent entanglement of condensates, like in a multigap BCS superconductor. Here the decoherence-evading qualities at high temperature are based on the Feshbach resonance that has been recently proposed as the driving mechanism for high T(c) superconductors. Finally we discuss how the proximity to a particular critical point is relevant to the emergence of coherence in the living cell. PMID- 19564943 TI - Cytotoxic effects of CdSe quantum dots on maturation of mouse oocytes, fertilization, and fetal development. AB - Quantum dots (QDs) are useful novel luminescent markers, but their embryonic toxicity is yet to be fully established, particularly in oocyte maturation and sperm fertilization. Earlier experiments by our group show that CdSe-core QDs have cytotoxic effects on mouse blastocysts and are associated with defects in subsequent development. Here, we further investigate the influence of CdSe-core QDs on oocyte maturation, fertilization, and subsequent pre- and postimplantation development. CdSe-core QDs induced a significant reduction in the rates of oocyte maturation, fertilization, and in vitro embryo development, but not ZnS-coated CdSe QDs. Treatment of oocytes with 500 nM CdSe-core QDs during in vitro maturation (IVM) led to increased resorption of postimplantation embryos and decreased placental and fetal weights. To our knowledge, this is the first study to report the negative impact of CdSe-core QDs on mouse oocyte development. Moreover, surface modification of CdSe-core QDs with ZnS effectively prevented this cytotoxicity. PMID- 19564945 TI - Stereodynamics study of the reaction of O(3P) with CH4(v = 0, j = 0). AB - A new London-Eyring-Polanyi-Sato (LEPS) potential energy surface (PES) is used in the O + CH(4) --> OH + CH(3) reaction via the quasiclassical trajectory method (QCT). Comparing with the experiments and the former ab initio calculations, the new LEPS PES describes the actual potential energy surface of the O + CH(4) reaction successfully. The four polarization dependent "generalized" differential cross sections (PDDCS) are presented in the center of mass frame. In the meantime, the distribution of dihedral angle [P(phi(r)), the distribution of angle between k and j' (P(theta(r))] and the angular distribution of product rotational vectors in the form of polar plots in theta(r) and phi(r) (P(theta(r), phi(r)) are calculated. The isotope effect for the reactions O + CD(4) is also calculated. These results are in good agreement with the experiments. PMID- 19564944 TI - The effect of self-assembling peptide RADA16-I on the growth of human leukemia cells in vitro and in nude mice. AB - Nanofiber scaffolds formed by self-assembling peptide RADA16-I have been used for the study of cell proliferation to mimic an extracellular matrix. In this study, we investigated the effect of RADA16-I on the growth of human leukemia cells in vitro and in nude mice. Self-assembly assessment showed that RADA16-I molecules have excellent self-assembling ability to form stable nanofibers. MTT assay displayed that RADA16-I has no cytotoxicity for leukemia cells and human umbilical vein endothelial cells (HUVECs) in vitro. However, RADA16-I inhibited the growth of K562 tumors in nude mice. Furthermore, we found RADA16-I inhibited vascular tube-formation by HUVECs in vitro. Our data suggested that nanofiber scaffolds formed by RADA16-I could change tumor microenvironments, and inhibit the growth of tumors. The study helps to encourage further design of self assembling systems for cancer therapy. PMID- 19564946 TI - Construction of an artificial MicroRNA expression vector for simultaneous inhibition of multiple genes in mammalian cells. AB - Recently, artificial microRNA (amiRNA) has become a promising RNA interference (RNAi) technology. Here, we describe a flexible and reliable method for constructing both single- and multi-amiRNA expression vectors. Two universal primers, together with two specific primers carrying the encoding sequence of amiRNA were designed and utilized to synthesize the functional amiRNA cassette through a one-step PCR. With appropriate restriction sites, the synthesized amiRNA cassettes can be cloned into any site of different destination vectors. Using the method, we constructed both single- and multi-amiRNA expression vectors to target three reporter genes, which code firefly luciferase (Fluc), enhanced green fluorescent protein (EGFP) and beta-galactosidase (LacZ), respectively. The expressions of three genes were all specifically inhibited by either the corresponding single- or the multi-amiRNA expression vector in 293T cells. And the RNAi efficiency of each amiRNA produced by both single- and multi-amiRNA expression vectors was comparable. PMID- 19564947 TI - Temperature-dependent phase behaviors in cylinder-forming block copolymers. AB - We demonstrate that the temperature-dependent phase behaviors of parallel and perpendicular cylinder-forming block copolymers are governed by domain-domain segregation forces inherently present in block copolymer material itself. With increasing temperature, a parallel cylinder-forming block copolymer experienced a parallel cylinder straightening process before the order-disorder transition (ODT) and did not show long-range composition fluctuations near the ODT temperature due to the weak segregation forces between the block domains. A perpendicular cylinder-forming block copolymer with a strong segregation force between the block domains displayed cylinder orientation transition from perpendicular to parallel below the ODT temperature. On the other hand, a perpendicular cylinder-forming block copolymer material with an exceptionally strong segregation force between the block domains maintained its initial perpendicular cylinder orientation up to near the ODT temperature. In both cases of perpendicular cylinder-forming block copolymers, submicrometer-scale long range composition fluctuations were observed well above the ODT temperature due to their intrinsically strong segregation forces between the block domains. PMID- 19564948 TI - Identifying protein-protein interaction sites using covering algorithm. AB - Identification of protein-protein interface residues is crucial for structural biology. This paper proposes a covering algorithm for predicting protein-protein interface residues with features including protein sequence profile and residue accessible area. This method adequately utilizes the characters of a covering algorithm which have simple, lower complexity and high accuracy for high dimension data. The covering algorithm can achieve a comparable performance (69.62%, Complete dataset; 60.86%, Trim dataset with overall accuracy) to a support vector machine and maximum entropy on our dataset, a correlation coefficient (CC) of 0.2893, 58.83% specificity, 56.12% sensitivity on the Complete dataset and 0.2144 (CC), 53.34% (specificity), 65.59% (sensitivity) on the Trim dataset in identifying interface residues by 5-fold cross-validation on 61 protein chains. This result indicates that the covering algorithm is a powerful and robust protein-protein interaction site prediction method that can guide biologists to make specific experiments on proteins. Examination of the predictions in the context of the 3-dimensional structures of proteins demonstrates the effectiveness of this method. PMID- 19564949 TI - Broad considerations concerning electrochemical electrodes in primarily fluid environments. AB - This review is variously a presentation, reflection, synthesis and report with reference to more recent developments of an article - in a journal which has ceased publication - entitled "Some Electrode Theorems with Experimental Corroboration, Inclusive of the Ag/AgCl System" Internet Journal of Chemistry, (http://www.ijc.com), Special Issues: Vol. 2 Article 24 (1999). The results from new lemmas relating charge densities and capacitance in a metallic electrode in equilibrium with an ionic solution are used to explain the data and observed effects due to Esin, Markov, Grahame, Lang and Kohn. Size effects that vary the measured e.m.f. of electrodes due to changes in the electronic chemical potential are demonstrated in experiment and theory implying the need for standardization of electrodes with respect to geometry and size. The widely used Stern modification of the Gouy-Chapman theory is shown to be mostly inapplicable for many of the problems where it is employed. Practical consequences of the current development include the possibility of determining the elusive single-ion activity coefficients of solution ions directly from the expression given by a simplified capacitance theorem, the potential of zero charge and the determination of single ion concentrations of active species in the electrode reactions from cell e.m.f. measurements. PMID- 19564951 TI - Elementary reactions and their role in gas-phase prebiotic chemistry. AB - The formation of complex organic molecules in a reactor filled with gaseous mixtures possibly reproducing the primitive terrestrial atmosphere and ocean demonstrated more than 50 years ago that inorganic synthesis of prebiotic molecules is possible, provided that some form of energy is provided to the system. After that groundbreaking experiment, gas-phase prebiotic molecules have been observed in a wide variety of extraterrestrial objects (including interstellar clouds, comets and planetary atmospheres) where the physical conditions vary widely. A thorough characterization of the chemical evolution of those objects relies on a multi-disciplinary approach: 1) observations allow us to identify the molecules and their number densities as they are nowadays; 2) the chemistry which lies behind their formation starting from atoms and simple molecules is accounted for by complex reaction networks; 3) for a realistic modeling of such networks, a number of experimental parameters are needed and, therefore, the relevant molecular processes should be fully characterized in laboratory experiments. A survey of the available literature reveals, however, that much information is still lacking if it is true that only a small percentage of the elementary reactions considered in the models have been characterized in laboratory experiments. New experimental approaches to characterize the relevant elementary reactions in laboratory are presented and the implications of the results are discussed. PMID- 19564952 TI - A novel method for the preparation of retinoic acid-loaded nanoparticles. AB - The goal of present work was to investigate the use of bioerodible polymeric nanoparticles as carriers of retinoic acid (RA), which is known to induce differentiation of several cell lines into neurons. A novel method, named "Colloidal-Coating", has been developed for the preparation of nanoparticles based on a copolymer of maleic anhydride and butyl vinyl ether (VAM41) loaded with RA. Nanoparticles with an average diameter size of 70 nm and good morphology were prepared. The activity of the encapsulated RA was evaluated on SK-N-SH human neuroblastoma cells, which are known to undergo inhibition of proliferation and neuronal differentiation upon treatment with RA. The activity of RA was not affected by the encapsulation and purification processes. PMID- 19564953 TI - Microscopic investigation of reversible nanoscale surface size dependent protein conjugation. AB - Abeta(1-40) coated 20 nm gold colloidal nanoparticles exhibit a reversible color change as pH is externally altered between pH 4 and 10. This reversible process may contain important information on the initial reversible step reported for the fibrillogenesis of Abeta (a hallmark of Alzheimer's disease). We examined this reversible color change by microscopic investigations. AFM images on graphite surfaces revealed the morphology of Abeta aggregates with gold colloids. TEM images clearly demonstrate the correspondence between spectroscopic features and conformational changes of the gold colloid. PMID- 19564955 TI - To what extent is water responsible for the maintenance of the life for warm blooded organisms? AB - In this work, attention is mainly focused on those properties of water which are essentially changed in the physiological temperature range of warm-blooded organisms. Studying in detail the half-width of the diffusion peak in the quasi elastic incoherent neutron scattering, the behavior of the entropy and the kinematic shear viscosity, it is shown that the character of the translational and rotational thermal motions in water radically change near T(H) ~ 315 K, which can be interpreted as the temperature of the smeared dynamic phase transition. These results for bulk pure water are completed by the analysis of the isothermic compressibility and the NMR-spectra for water-glycerol solutions. It was noted that the non-monotone temperature dependence of the isothermic compressibility (beta(T)) takes also place for the water-glycerol solutions until the concentration of glycerol does not exceed 30 mol%. At that, the minimum of beta(T) shifts at left when the concentration increases. All these facts give us some reasons to assume that the properties of the intracellular and extracellular fluids are close to ones for pure water. Namely therefore, we suppose that the upper temperature limit for the life of warm-blooded organisms [T(D) = (315 +/- 3) K] is tightly connected with the temperature of the dynamic phase transition in water. This supposition is equivalent to the assertion that the denaturation of proteins at T > or = T(H) is mainly provoked by the rebuilding of the H-bond network in the intracellular and extracellular fluids, which takes place at T > or = T(H). A question why the heavy water cannot be a matrix for the intracellular and extracellular fluids is considered. The lower physiological pH limit for the life of warm-blooded organisms is discussed. PMID- 19564954 TI - Hypoglycaemic and hypolipidaemic effects of Withania somnifera root and leaf extracts on alloxan-induced diabetic rats. AB - Withania somnifera is an important medicinal plant, which is used in traditional medicine to cure many diseases. Flavonoids were determined in the extracts of W. somnifera root (WSREt) and leaf (WSLEt). The amounts of total flavonoids found in WSREt and WSLEt were 530 and 520 mg/100 g dry weight (DW), respectively. Hypoglycaemic and hypolipidaemic effects of WSREt and WSLEt were also investigated in alloxan-induced diabetic rats. WSREt and WSLEt and the standard drug glibenclamide were orally administered daily to diabetic rats for eight weeks. After the treatment period, urine sugar, blood glucose, haemoglobin (Hb), glycosylated haemoglobin (HbA1C), liver glycogen, serum and tissues lipids, serum and tissues proteins, liver glucose-6-phosphatase (G6P) and serum enzymes like aspartate transaminase (AST), alanine transaminase (ALT), acid phosphatase (ACP) and alkaline phosphatase (ALP) levels were determined. The levels of urine sugar, blood glucose, HbA1C, G6P, AST, ALT, ACP, ALP, serum lipids except high density lipoprotein-bound cholesterol (HDL-c) and tissues like liver, kidney and heart lipids were significantly (p < 0.05) increased, however Hb, total protein, albumin, albumin:globulin (A:G) ratio, tissues protein and glycogen were significantly (p < 0.05) decreased in alloxan-induced diabetic rats. Treatment of the diabetic rats with WSREt, WSLEt and glibenclamide restored the changes of the above parameters to their normal level after eight weeks of treatment, indicating that WSREt and WSLEt possess hypoglycaemic and hypolipidaemic activities in alloxan-induced diabetes mellitus (DM) rats. PMID- 19564957 TI - Analysis of intestinal injuries induced by ricin in vitro using SPR technology and MS identification. AB - The present study found that ricin toxicity did not only manifest itself as inhibition of protein synthesis, but also induced apoptosis of immune cells and played an extremely significant role in intestinal injury. In this report, we describe a novel method to estimate binding events occurring on intestinal brush border membranes (BBM) based on SPR technology in an attempt to mimic the real intestinal surface capable of interacting physically and/or actively with certain biological molecules. Combined with HPCE-ESI-MS indentification, we obtained 28 kinds of proteins in BBM that interacted with ricin. PMID- 19564956 TI - Relative stabilities of conserved and non-conserved structures in the OB-fold superfamily. AB - The OB-fold is a diverse structure superfamily based on a beta-barrel motif that is often supplemented with additional non-conserved secondary structures. Previous deletion mutagenesis and NMR hydrogen exchange studies of three OB-fold proteins showed that the structural stabilities of sites within the conserved beta-barrels were larger than sites in non-conserved segments. In this work we examined a database of 80 representative domain structures currently classified as OB-folds, to establish the basis of this effect. Residue-specific values were obtained for the number of Calpha-Calpha distance contacts, sequence hydrophobicities, crystallographic B-factors, and theoretical B-factors calculated from a Gaussian Network Model. All four parameters point to a larger average flexibility for the non-conserved structures compared to the conserved beta-barrels. The theoretical B-factors and contact densities show the highest sensitivity. Our results suggest a model of protein structure evolution in which novel structural features develop at the periphery of conserved motifs. Core residues are more resistant to structural changes during evolution since their substitution would disrupt a larger number of interactions. Similar factors are likely to account for the differences in stability to unfolding between conserved and non-conserved structures. PMID- 19564950 TI - Mitochondria and energetic depression in cell pathophysiology. AB - Mitochondrial dysfunction is a hallmark of almost all diseases. Acquired or inherited mutations of the mitochondrial genome DNA may give rise to mitochondrial diseases. Another class of disorders, in which mitochondrial impairments are initiated by extramitochondrial factors, includes neurodegenerative diseases and syndromes resulting from typical pathological processes, such as hypoxia/ischemia, inflammation, intoxications, and carcinogenesis. Both classes of diseases lead to cellular energetic depression (CED), which is characterized by decreased cytosolic phosphorylation potential that suppresses the cell's ability to do work and control the intracellular Ca(2+) homeostasis and its redox state. If progressing, CED leads to cell death, whose type is linked to the functional status of the mitochondria. In the case of limited deterioration, when some amounts of ATP can still be generated due to oxidative phosphorylation (OXPHOS), mitochondria launch the apoptotic cell death program by release of cytochrome c. Following pronounced CED, cytoplasmic ATP levels fall below the thresholds required for processing the ATP-dependent apoptotic cascade and the cell dies from necrosis. Both types of death can be grouped together as a mitochondrial cell death (MCD). However, there exist multiple adaptive reactions aimed at protecting cells against CED. In this context, a metabolic shift characterized by suppression of OXPHOS combined with activation of aerobic glycolysis as the main pathway for ATP synthesis (Warburg effect) is of central importance. Whereas this type of adaptation is sufficiently effective to avoid CED and to control the cellular redox state, thereby ensuring the cell survival, it also favors the avoidance of apoptotic cell death. This scenario may underlie uncontrolled cellular proliferation and growth, eventually resulting in carcinogenesis. PMID- 19564958 TI - Problems of prescription drug use in children. PMID- 19564959 TI - Off-label use of prescription drugs in childhood and adolescence: an analysis of prescription patterns in Germany. AB - BACKGROUND: Not all drugs prescribed for children and adolescents are certified for use in these age groups. The present study describes the extent of off-label prescriptions in the outpatient sector and identifies deficits in the quality of product information. METHODS: This analysis is based on patient-specific but pseudonymized drug prescription data for approximately 289,000 0- to 16-year-old members of the Gmunder Ersatzkasse, a German statutory health insurance provider, in the year 2002. For each substance prescribed, information regarding the youngest certified age group was derived from the product data. RESULTS: Of the 1,429,981 prescribed drug packages (726 active substances), 87.4% (66.1% of active substances) were prescribed in accordance with their license and 3.2% (15.7% of active substances) were prescribed off-label. For 9.4% of prescriptions (18.2% of active substances) the licensing status could not be established. For neonates and infants the proportion of licensed prescriptions was below average, at 42.5% and 82.8% of the prescribed packages (20.0% and 38.6% of active substances) respectively. After infancy, prescriptions were predominantly in accordance with licensed use. Deficits were seen in the indication groups "alimentary tract and metabolism," "respiratory system," "dermatologicals," and "sensory organs." CONCLUSIONS: The methodology enables characterization of off label prescription and identification of fields where further research is needed. With regard to the EU regulation on medicinal products for pediatric use, this could assume increasing importance and contribute to the development of appropriate and safe medicines for children. PMID- 19564961 TI - Living with a dilemma. PMID- 19564960 TI - Hyperhidrosis--causes and treatment of enhanced sweating. AB - BACKGROUND: Basically two types of sweating exist: thermoregulatory and emotional sweating. They are controlled by different centers: thermo regulatory sweating is regulated predominantly by the hypothalamus, emotional sweating predominantly by the limbic system. Enhanced sweating, called hyperhidrosis, can be generalized or focal. Primary focal hyperhidrosis is the most common type and affects the axillae, hands, feet, and face--areas principally involved in emotional sweating. Secondary hyperhidrosis develops due to dysfunction of the central or peripheral nervous system. METHODS: Review based on a selective search of the literature via Medline and on the guidelines of the Association of the Scientific Medical Societies in Germany (Arbeitsgemeinschaft der wissenschaftlichen medizinischen Fachgesellschaften [AWMF]). RESULTS: Various conservative and surgical treatments exist for hyperhidrosis. Conservative treatment options are the local application of aluminum chloride, tap water iontophoresis, and the intracutaneous injection of botulinum toxin. Surgical approaches include endoscopic sympathectomy and axillary tumescent curettage and liposuction, removing the sweat glands. Systemic drugs (e.g. anticholinergic substances) can be used in the treatment of generalized hyperhidrosis. CONCLUSION: A step-by-step approach is recommended for the treatment of hyperhidrosis. Local treatment options with few and minor side effects should be tried first. PMID- 19564962 TI - Back orthoses are important. PMID- 19564963 TI - Impairments precede fractures. PMID- 19564964 TI - Unreliable in the individual case. PMID- 19564966 TI - Cardiac gene defects can cause sudden cardiac death in young people. AB - BACKGROUND: In Europe, sudden cardiac death (SCD) is one of the most common causes of death. Although sudden cardiac death usually happens in older people, 5% to 10% of the affected individuals are young and apparently healthy. Sudden death in infants, children, and young adults is relatively rare, with an incidence of 1 to 5 per 100 000 persons per year. Nonetheless, up to 7000 asymptomatic children die in the USA each year, almost half of them without any warning signs or symptoms. METHOD: Selective literature review. RESULTS: Although structural cardiovascular abnormalities explain most cases of sudden cardiac death in young people, the cause of death remains unexplained after autopsy in 10% to 30% of cases. Potentially lethal ion channel disorders (channelopathies) such as the long QT syndromes (LQTS), catecholaminergic polymorphic ventricular tachycardia (CPVT), and the Brugada syndrome (BrS) may account for at least one third of these unexplained cases. Most of these diseases are hereditary with autosomal-dominant transmission, i.e., there is a 50% chance that the children of affected individuals will be affected themselves. CONCLUSIONS: Post-mortem genetic screening for sequence variations in cardiac ion channel genes has become an important forensic tool for elucidating the cause of sudden cardiac death. Moreover, it allows the identification of other family members bearing the previously undiagnosed gene defect, who can then undergo a cardiological evaluation if indicated by their clinical history. PMID- 19564968 TI - Advise against metamizole. PMID- 19564967 TI - Point-of-care testing in microbiology: the advantages and disadvantages of immunochromatographic test strips. AB - BACKGROUND: Point-of-care testing (POCT) for the demonstration of pathogens was introduced several years ago. The present study describes the current technical status of POCT, giving some examples, and summarizes the specific advantages and disadvantages of the POCT approach in microbiology. METHODS: Selective review of the literature found in medical databases under consideration of current German and international guidelines. RESULTS/CONCLUSIONS: The test systems available today are technically mature and offer good to very good performance. For HIV, malaria, group A streptococci, and legionellae, POCT testing, when indicated, is on a par with conventional procedures. The information yielded by rapid tests for pneumococci and for influenza tends to be supplementary in nature. The rapid test for group B streptococci is unsuitable for routine use because its sensitivity is still too low compared with bacterial culture. POCT can be successful only if the tests are performed correctly by trained personnel, quality management procedures are followed, and the severity of illness and the epidemiological circumstances are taken into account when interpreting the results. PMID- 19564969 TI - Current data situation. PMID- 19564970 TI - Mention the ophthalmologist. PMID- 19564973 TI - At a loss. PMID- 19564974 TI - Cautious advice. PMID- 19564975 TI - Selection of Pneumococci. PMID- 19564977 TI - General practitioners need support. PMID- 19564978 TI - AGnES: supporting general practitioners with qualified medical practice personnel: model project evaluation regarding quality and acceptance. AB - BACKGROUND: The German AGnES (community-based, e-health-assisted systemic support for primary care) project allows general practitioners (GPs) to delegate certain elements of medical care, including house calls, to qualified AGnES employees and thereby provide primary care to a larger number of patients. AGnES projects of various types have been carried out in a number of German federal states from 2005 onward. In this article, an evaluation of the AGnES projects to date is presented. METHODS: Patient data (age, sex, diagnoses, level of care, mobility, etc.) and each of the specific activities carried out in the AGnES framework were documented with standardized computer-based instruments. The GPs, AGnES employees, and patients also underwent standardized interviews. The acceptance of the AGnES project, competence of the AGnES employees, and quality of medical care within the projects were evaluated. The participating GPs themselves assessed the quality of medical care. RESULTS: By July 8, 2008, 8386 house calls on a total of 1486 patients had been made within the framework of the AGnES projects. The evaluation revealed a high degree of acceptance of the project among the participating GPs, AGnES employees, and patients. The GPs considered the quality of medical care within the AGnES project to be good for the vast majority of patients. CONCLUSION: Structural redundancy is avoided by directly placing the AGnES employees in the general practitioners' practices. Based on the results of the AGnES projects, the law in Germany has now been amended to enable implementation of the AGnES project in the regular health care system from January 2009 onward. The next steps to be taken are the establishment of adequate reimbursement within the catalog of the statutory health insurance scheme and a detailed definition of the required qualifications. PMID- 19564979 TI - Maturation of the auditory pathway. PMID- 19564981 TI - Monocausal discussion. PMID- 19564983 TI - Why did I do it? Would I do it again? A 30-year homage. PMID- 19564982 TI - Psoriasis--new insights into pathogenesis and treatment. AB - BACKGROUND: Psoriasis is one of the most prevalent chronic inflammatory diseases, affecting approximately 2 million people in Germany. METHODS: Selective literature review taking into account the German S1 and S3 guidelines for the treatment of this condition. RESULTS AND CONCLUSIONS: Psoriasis is a very troublesome disease with a high economic impact. The disease often persists for life, and the patient has an increased risk of cardiovascular diseases and their complications. One out of five patients develops psoriatic arthritis. The clinical picture of psoriasis is highly variable with regard to lesional characteristics and the severity of disease. To improve the management of psoriasis the guidelines must be followed and all appropriate topical and systemic treatment options must be tried, with clearly defined treatment goals. The spectrum of established systemic treatments for psoriasis has been extended by the biologics. These can be used to achieve a good skin status and a clear-cut improvement in quality of life even in patients who do not--or no longer--respond adequately to conventional therapies. PMID- 19564984 TI - New IDEAS to improve the quality of AJPE articles. PMID- 19564985 TI - Effect of learner-centered teaching on motivation and learning strategies in a third-year pharmacotherapy course. AB - OBJECTIVES: To develop, implement, and assess a learner-centered approach to teaching a third-year pharmacotherapy course in a doctor of pharmacy (PharmD) program. METHODS: The pharmacotherapy course was restructured according to the learner-centered approach. The Motivated Strategies for Learning Questionnaire (MSLQ) was administered to students before and after taking the course, and changes in MSLQ subscales from baseline were evaluated. Students' response to the learner-centered approach and characteristics associated with MSLQ scores were also evaluated. RESULTS: Compared to baseline, students' intrinsic goal orientation control of learning beliefs, self-efficacy, critical thinking, and metacognitive self-regulation improved after taking the course. Students responded positively to the learner-centered approach. Additionally, students with a clinical practice career orientation or who prepared frequently for classes scored higher on several MSLQ domains. CONCLUSIONS: The learner-centered approach was effective in promoting several domains of motivation and learning strategies in a third-year pharmacotherapy course. PMID- 19564986 TI - A genotyping exercise for pharmacogenetics in pharmacy practice. AB - OBJECTIVE: To develop a genotype exercise to improve pharmacy students' comprehension of pharmacogenetic principles that apply to patient care. DESIGN: Deoxyribonucleic acid (DNA) was collected during class from 10 student volunteers and subjected to genotype analysis. The results were presented to the class and discussed in the context of a patient genetic counseling session. Students completed a survey instrument regarding their attitudes toward this learning experience. ASSESSMENT: Students indicated that the exercise engaged them with the course content and would positively influence their ability to apply pharmacogenetic principles to patient care. CONCLUSION: An applied genotype exercise enhanced learning of pharmacogenetic principles. Based on these findings, conducting a genotype exercise in a large classroom setting is feasible in terms of time and expense, and meaningful in terms of student satisfaction. PMID- 19564987 TI - A hospice-based advanced pharmacy experience. AB - OBJECTIVE: To evaluate a hospice-based advanced pharmacy practice experience (APPE). DESIGN: Two years of data gathered from student evaluation forms and reflective journals were analyzed. ASSESSMENT: Students completed reflective journals and expressed a high level of satisfaction with the hospice-based learning experience. They gained a better understanding of end-of-life care provided by a hospice and the pharmacist's role in optimizing supportive care for patients receiving hospice care. CONCLUSION: Hospice-based APPEs can provide a rich interdisciplinary learning environment for pharmacy students interested in developing knowledge, attitudes, and skills to effectively manage the pharmacotherapy of patients receiving end-of-life care. PMID- 19564988 TI - Instructor satisfaction with a technology-based resource for diabetes education. AB - OBJECTIVES: To evaluate instructor use patterns and satisfaction with DM Educate, a comprehensive, Web-based diabetes course. METHODS: Instructors completed a post course survey instrument to assess their use of course materials and components, as well as satisfaction with the course content, design, and technology utilized, and to solicit their suggestions for additional content areas. RESULTS: Thirty eight percent of respondents utilized DM Educate as a standalone elective and 62% had integrated materials into existing courses. The pharmacotherapy module was the most utilized at 91% and slide sets were the most utilized course components at 63%. All instructors stated that they would use the course again the following year. Suggestions for improvement included incorporation of more active-learning activities and patient cases. CONCLUSION: Instructors' were highly satisfied with the course materials and technology used by DM Educate, a Web-based diabetes education course, and indicated they were able to customize the course materials both to establish new courses and supplement existing courses. All instructors planned to use the course again. PMID- 19564989 TI - Economic impact of pharmacy graduates on a regional economy. AB - OBJECTIVES: To analyze the impact of recent pharmacy graduates on a local economy. METHODS: Input-output analysis was applied to data from Spokane County, Washington, in 2006 and the findings were reviewed and conclusions were drawn. RESULTS: The local college of pharmacy added nearly $1 million (in 2006) directly to the local economy. New pharmacists added nearly $400,000 in direct value. However, because the graduates alleviated a shortage of pharmacists in the area, thereby avoiding both the tangible and intangible (eg, human health) economic costs of a continued shortage, the true economic impact may have been even greater. CONCLUSIONS: Doctor of pharmacy (PharmD) graduates entering the workforce add substantial value, both to the local retail pharmacy industry specifically and the local economy in general. Thus, the economic impact of the pharmacy practice program training these students is also substantial. PMID- 19564990 TI - Preceptor perceptions of pharmacy student team quality assurance projects. AB - OBJECTIVES: To assess preceptors' opinions of the impact of quality assurance projects. METHODS: Students were given the opportunity to directly apply material learned in class in a "real world" environment by completing a quality assurance project in a community or health-system pharmacy. All preceptors (n = 38) were contacted via telephone and given the opportunity to respond to open-ended questions concerning their experience with student-team quality assurance projects. RESULTS: Preceptors indicated the quality assurance projects benefited their practice sites by providing additional resources (53%, n = 19), decreased medication errors (22%, n = 8), and increased awareness of the importance of quality assurance (22%, n = 8). Ninety-four percent of respondents (n = 34) perceived the projects had a positive impact on patient care and 92% (n = 33) perceived a positive impact on themselves. CONCLUSIONS: Preceptors felt that quality assurance projects performed by pharmacy-student teams were beneficial to patient care, the practice site, and themselves. The quality assurance projects have broad applications and can be added to a medication safety class or to the introductory pharmacy practice experience (IPPE) sequence. PMID- 19564991 TI - Student leadership retreat focusing on a commitment to excellence. AB - OBJECTIVE: To design and evaluate a student leadership retreat focused on: leadership regardless of position or title, the need for passionate commitment to excellence, the importance of teamwork, and the value of self-reflection. DESIGN: Students in their second, third, and fourth year of a doctor of pharmacy (PharmD) program were invited to participate in an off-campus leadership retreat that combined organized learning activities and social/networking opportunities. Retreat content utilized didactic, experiential, and self-directed learning activities. ASSESSMENT: Forty-two students participated in the retreat and were surveyed pre- and post-retreat. A student-led group evaluation and course evaluations were also utilized. Responses on the post-retreat evaluation suggested an increased belief in their ability to influence change and an enhanced awareness of the role of self-reflection in creating excellence in the profession. CONCLUSION: An off-campus overnight retreat has value in challenging student perceptions about leadership and promoting reflection on their personal role in leading change within the profession Continued exploration and development of this format is warranted for commitment to excellence programming and also for additional leadership topics and audiences. PMID- 19564992 TI - A hybrid jigsaw approach to teaching renal clearance concepts. AB - OBJECTIVES: To evaluate the effectiveness of using a jigsaw cooperative learning approach to teach basic concepts of renal clearance to pharmacy students. DESIGN: Students collected information on the mechanisms of renal clearance for a particular drug and proposed a methodology for circumventing a urine drug screen. Attitudinal surveys, an online quiz, and course examinations were used to assess student learning. ASSESSMENT: The majority of students felt apprehensive toward a group assignment prior to the exercise, and afterwards still preferred individual work over group work. Post-exercise quiz and final examination scores showed students successfully learned the material. CONCLUSIONS: Students were successful in learning from each other and there was no difference in examination performance compared to years when the technique was not used. In addition, the relative negative experiences of previous group work decreased the subjective attitudes related to the current learning experience. PMID- 19564993 TI - Integration of first- and second-year introductory pharmacy practice experiences. AB - OBJECTIVES: To describe the integration of a first- and second-year introductory pharmacy practice experience (IPPE) involving direct patient contact in hospitals and clinics as a means of more efficiently using academic and preceptor resources. DESIGN: Two IPPE courses were integrated in fall 2004 to accommodate increasing enrollment in classes and limited clinical practice sites and preceptors, as well as to meet the increased need for students and clinicians to practice principles of self-education. P1 and P2 students interviewed patients and presented patient cases; preceptor expectations were structured by instructional objectives. Student and preceptor course evaluations were assessed from survey data. ASSESSMENT: During the assessment period, all students passed the courses. Following integration of the IPPEs, both courses received positive evaluations from students and preceptors. Initial advanced pharmacy practice experience (APPE) grades for students completing the courses further suggests that the integrated IPPEs were beneficial to students. CONCLUSION: The successful integration of first- and second- year IPPE courses resulted in more efficient use of academic and preceptor resources and created a model for other colleges of pharmacy to consider. PMID- 19564994 TI - An advanced cardiovascular pharmacotherapy course blending online and face-to face instruction. AB - OBJECTIVE: To assess the effectiveness of online instruction in a cardiology pharmacotherapy elective. DESIGN: Eight drug-focused lectures and 6 introductory presentations were added to a cardiology pharmacotherapy course. Students completed an online quiz after each online drug-focused lecture and scores were compared to quizzes taken at the beginning and end of the course, as well as on a cardiology advanced pharmacy practice experience (APPE). For online introductory presentations, students completed a quiz at the beginning of the next face-to face session. A survey was conducted at the end of the course to obtain student feedback. ASSESSMENT: Compared to baseline scores, student learning was demonstrated after online drug-focused lectures by higher quiz scores attained immediately after completing the lecture, at the end of the course, and at the beginning of the APPE. Furthermore, students performed better on quizzes at the beginning of face-to-face sessions if they first completed an online introductory presentation. Students expressed strong support for the online components of the course. CONCLUSIONS: A blended learning environment with online and face-to-face instruction is an effective way to teach a cardiology pharmacotherapy elective. The online component of this course was well received by students, improved student preparation before attending class, and appeared to enhance long-term cardiovascular drug knowledge. PMID- 19564995 TI - Pharmacists' and pharmacy students' ability to identify drug-related problems using TIMER (Tool to Improve Medications in the Elderly via Review). AB - OBJECTIVE: Determine the effectiveness of TIMER (Tool to Improve Medications in the Elderly via Review) in helping pharmacists and pharmacy students identify drug-related problems during patient medication reviews. METHODS: In a randomized, controlled study design, geriatric patient cases were sent to 136 pharmacists and 108 third-year pharmacy students who were asked to identify drug related-problems (DRPs) with and without using TIMER. RESULTS: Pharmacists identified more tool-related DRPs using TIMER (p = 0.027). Pharmacy students identified more tool-related DRPs using TIMER in the first case (p = 0.02), but not in the second. CONCLUSION: TIMER increased the number of DRPs identified by practicing pharmacists and pharmacy students during medication reviews of hypothetical patient cases. PMID- 19564996 TI - Medicare part D community outreach train-the-trainer program for pharmacy faculty. AB - OBJECTIVES: To assess the train-the-trainer component of an initiative (Partners in D) to train pharmacy students to facilitate patient enrollment in the best Medicare Part D prescription drug plan (Part D). METHODS: Faculty members from 6 California colleges or schools of pharmacy were taught how to train pharmacy students about Medicare Part D and how to conduct outreach events targeting underserved patient populations. A preintervention and postintervention survey instrument was administered to determine participants' (1) knowledge of the Part D program; (2) skill using the Medicare Prescription Drug Plan Finder tool; and (3) confidence in their ability to train pharmacy students. Implementation of the Partners in D curriculum in faculty members' colleges or schools of pharmacy was also determined. RESULTS: Participants' knowledge of Part D, mastery of the Plan Finder, and confidence in teaching the material to pharmacy students all significantly improved. Within 8 weeks following the program, 5 of 6 colleges or schools of pharmacy adopted Partners in D coursework and initiated teaching the Partners-in-D curriculum. Four months afterwards, 21 outreach events reaching 186 Medicare beneficiaries had been completed. CONCLUSIONS: The train-the-trainer component of the Partners in D program is practical and effective, and merits serious consideration as a national model for educating patients about Medicare Part D. PMID- 19564997 TI - Independent community pharmacists' perspectives on compounding in contemporary pharmacy education. AB - OBJECTIVES: To identify compounding practices of independent community pharmacy practitioners in order to make recommendations for the development of curricular objectives for doctor of pharmacy (PharmD) programs. METHODS: Independent community practitioners were asked about compounding regarding their motivations, common activities, educational exposures, and recommendations for PharmD education. RESULTS: Most respondents (69%) accepted compounding as a component of pharmaceutical care and compounded dermatological preparations for local effects, oral solutions, and suspensions at least once a week. Ninety-five percent were exposed to compounding in required pharmacy school courses and most (98%) who identified compounding as a professional service offered in their pharmacy sought additional postgraduate compounding education. Regardless of the extent of compounding emphasis in the practices surveyed, 84% stated that PharmD curricula should include compounding. CONCLUSIONS: Pharmacy schools should define compounding curricular objectives and develop compounding abilities in a required laboratory course to prepare graduates for pharmaceutical care practice. PMID- 19564998 TI - Updated guidelines for manuscripts describing instructional design and assessment: the IDEAS format. PMID- 19564999 TI - Improving pharmacy students' education through enhanced experiential learning. PMID- 19565000 TI - South Asian subcontinent not represented in international pharmacy education supplement. PMID- 19565001 TI - Look in the looking glass, not through it. PMID- 19565002 TI - Tuberculosis: A re-emerging enemy. PMID- 19565003 TI - Rat activin-betaE mRNA expression during development and in acute and chronic liver injury. AB - Activin-betaE mRNA expression was investigated in male and female rats using gel based and quantitative RT-PCR, in fetal and post-natal liver during development and in a variety of tissues from 200 gm adult animals. Activin-betaE expression was also assessed in rat liver after partial hepatectomy, and after repeated toxic insult. Male Sprague Dawley rats were subjected to partial hepatectomy or sham operations. Samples were collected from the caudate liver lobe during regeneration, from 12 to 240 hr after surgery. Three groups of 5 male rats were treated with CCl(4) for 0 (control), 5 or 10 weeks, to induce liver fibrosis and cirrhosis. Activin-betaE mRNA was predominantly expressed in liver, with much lower amounts of mRNA observed in pituitary, adrenal gland and spleen, in both males and females. Low activin-betaE expression was observed in liver at fetal day 16, with higher levels seen between post-natal days 3 and 35 and a further increase noted by day 47, in both males and females. Liver activin-betaE mRNA concentrations did not change from control values 12-72 hr after PHx, but significantly increased over six fold, 168 hr post-hepatectomy, when liver mass was restored. Activin-betaE mRNA was up-regulated after 5 weeks of CCl(4) treatment, but not after 10 weeks. The changes in activin-betaE mRNA concentrations after liver insult did not always parallel those reported for the activin-betaC subunit, suggesting activin-betaE may have an independent role in liver under certain conditions. PMID- 19565005 TI - Showcasing the developing world. PMID- 19565004 TI - Retroviral vectors containing Tet-controlled bidirectional transcription units for simultaneous regulation of two gene activities. AB - In this study retroviral self-inactivating (SIN)-vectors were constructed, that allow simultaneous regulation of two genes by integration of bidirectional Tet controlled transcription units. Marker genes (luciferase and eGFP) were expressed under the control of various bidirectional promoters P(tet)bis, in order to determine (i) the fraction of HtTA-1 cells exhibiting tight doxycycline (Dox) dependent control; (ii) possible effects of the vector backbone on the regulation of gene transcription; (iii) the possibility for crosstalk between different minimal promoters within P(tet)bi. When HtTA-1 cells, constitutively expressing the Tet-Transactivator (tTA), were transduced by S2f-lMCg retroviral vector, a high percentage (40) of the cell population displayed tight regulation (5000 fold) of P(tet)bi activity over a wide range of Dox concentrations. As a result of our comparative study on the activity of virus derived minimal promoters (from MMTV, HIV and CMV), a clear hierarchy of activity as well as a different sensitivity to external influences among the various promoters studied was observed. Furthermore, our results strongly support the idea, that viral elements such as part of the MuLV pol/env region significantly affect the regulation capacity of an integrate. Taking into account our observations as outlined above, we succeeded in generating significantly optimized Tet regulated retroviral vectors. The application of such a one-step transfer system for P(tet) controlled genes would be of particular relevance to applications where cellular systems do not allow prolonged selection procedures as it is the case with primary cells considered for ex vivo gene therapy. PMID- 19565006 TI - An arylsulphatase A (ARSA) frameshift mutation (289insG) in metachromatic leukodystrophy (MLD). PMID- 19565007 TI - Polymorphism of the Fcgamma receptor IIA and malaria morbidity. AB - Fc receptors (FcRs) are expressed on the surface of all types of cells of the immune system. They bind the Fc portion of immunoglobulin (Ig), thereby bridging specific antigen recognition by antibodies with cellular effector mechanisms. FcgammaRIIA, one of the three receptors for human IgG, is a low-affinity receptor for monomeric IgG, but binds IgG immune complexes efficiently. FcgammaRIIA is believed to play a major role in eliciting monocyte- and macrophage-mediated effector responses against blood-stage malaria parasites. A G --> A single nucleotide polymorphism, which causes an arginine (R) to be replaced with histidine (H) at position 131, defines two allotypes which difer in their avidity for complexed human IgG(2) and IgG(3). Because FcgammaRIIA-H131 is the only FcgammaR allotype which interacts efficiently with human IgG(2,) this polymorphism may determine whether parasite-specific IgG(2) may or may not elicit cooperation with cellular imune responses during blood-stage malaria infection. Here, we review data from four published case-control studies describing associations between FcgammaRIIA R/H131 polymorphism and malaria-related outcomes and discuss possible reasons for some incongruities found in these available results. PMID- 19565009 TI - Docosahexaenoic acid inhibits protein kinase C translocation/activation and cardiac hypertrophy in rat cardiomyocytes. AB - Phenylephrine (PE) induces cardiac hypertrophy through multiple signaling pathways including pathways involving protein kinase C (PKC) activation. Docosahexaenoic acid (DHA), an omega-3 fatty acid, has been shown to reduce the PE-induced hypertrophic responses. However, the effects of DHA on PKC activation and translocation are controversial. The present study investigates the effect of DHA on PE-induced activation of PKC. The results indicate that PE induces PKCalpha translocation (from cytosol to plasma membranes) and activation in cardiomyocytes during the hypertrophic responses. Although DHA itself has no significant effect on basal PKC translocation and activation, it effectively reduced PE-stimulated PKC translocation and activation. The results of the present study suggest a possible mechanism explaining how dietary fish oil may inhibit development of cardiac hypertrophy and therefore may be an attractive dietary agent for preventing cardiac hypertrophy in patients with heart failure. PMID- 19565010 TI - Lack of association of methylenetetrahydrofolate reductase 677C>T mutation with coronary artery disease in a Pakistani population. AB - Pakistanis belong to the South Asian population which has the highest known rate of coronary artery disease. Folic acid deficiency also appears to be highly prevalent in this population. Methylenetetrahydrofolate reductase (MTHFR) 677C>T polymorphism decreases the activity of this enzyme and can be associated with mild to moderate hyperhomocysteinemia in homozygotes, particularly when there is folic acid deficiency, as well as with coronary artery disease. To assess the value of genotyping the MTHFR 677C>T dimorphism, we carried out a case-control study of dimorphism 677C>T for putative association with myocardial infarction (MI) among Pakistani nationals. We investigated a sample population of 622 Pakistanis consisting of 225 controls and 397 patients with clinical diagnosis of acute MI (AMI). MTHFR C677T alleles were determined by assays based on polymerase chain reaction and restriction endonuclease analysis. Frequencies of C alleles were 0.87 among controls and 0.86 among AMI patients. The MTHFR 677C>T dimorphism showed no association with MI (chi(2) = 0.25, 1df, P=0.62), serum levels of folate and vitamin B12 and plasma level of vitamin B6. A significant association, however, was found between homozygous 677T genotype and plasma levels of homocysteine. Multivariate analysis of the data showed that in case of log homocysteine, age and MTHFR genotypes were significantly different (P<0.001). In case of B12, smoking and age were found to be statistically significant (P<0.001), while in case of serum folate only smoking was found to be significant (P<0.001). The results indicate that MTHFR 677C>T polymorphism, though associated with homocysteine levels, confers no significant risk of coronary artery disease in the Pakistani population investigated here. We suggest that the higher incidence of AMI in South Asia occurs through mechanisms other than the MTHFR related pathways. PMID- 19565008 TI - Significance of the Y-box proteins in human cancers. AB - Y-box proteins belong to the cold shock domain family of proteins that are known to be involved in both transcriptional and translational control. Here, we give a brief overview of the structure, regulation and physiological functions of the Y box proteins. This is followed by examining the role of Y-box protein 1 (YB-1), the most extensively studied of the Y-box protein in tumorigenesis, and its clinicopathological significance. YB-1 has the potential to be a prognostic marker and predictor of chemoresistance in human cancers. PMID- 19565012 TI - Elements of a flu pandemic. PMID- 19565011 TI - Progesterone inhibition of MDM2 p90 protein in MCF-7 human breast cancer cell line is dependent on p53 levels. AB - The mdm2 gene encodes several protein isoforms with different molecular weights (p90, p80, p76 and p57). MDM2 p90 (usually considered to be the major MDM2 protein) binds to and inactivates P53. We have recently shown that growth inhibition of MCF-7 human breast cancer cells by progesterone is associated with P53 down-regulation. In this work, we analyzed the expression pattern of MDM2 proteins in three human breast cancer cell lines by western blotting with anti MDM2 antibodies. We found a prominent expression of MDM2 p57 protein in cell lines which have non-functional P53 protein (T47D and MDA-MB-231) as compared to the p90, p80 isoforms, whereas p90 was the major protein isoform in MCF-7 cells that contain functional P53 protein. When MCF-7 cells were treated with 100 nM of progesterone, MDM2 p90 was inhibited but the highly expressed MDM2 p57 isoform was not. The inhibition of MDM2 p90 protein by progesterone was abrogated in MCF 7 cells transfected with a P53 expressing vector. To our knowledge, this is the first report linking progesterone-induced growth inhibition with down-regulation of the MDM2 protein. We present evidence that reestablishing of P53 expression by transient transfection of P53 cDNA in these cells enhances the expression level of MDM2 p90 isoform. The data indicate that expression of MDM2 p90 is regulated through a P53-dependent pathway in response to progesterone. PMID- 19565013 TI - Population stratification and genetic association studies in South Asia. AB - Population stratification and its influence on genetic association studies is a controversial topic. Although it has been suggested that stratification is unlikely to bias the results of association studies conducted in developed countries, convincing contrary empirical evidence has been published. However, it is in populations where historical ethnic, religious and language barriers exist that community subdivisions will predictably exert greatest genetic effect, and influence the organization of association studies. In many of the populations of the Indian sub-continent, these basic population divisions are compounded by a strict tradition of intra-community marriage and by marriage between close biological relatives. Data on the very significant levels of genetic diversity that characterize the populations of India and Pakistan, with some 50,000-60,000 caste and non-caste communities in India, and average first cousin marriage rates of 40%-50% in Pakistan, are presented and discussed. Under these circumstances, failure to explicitly control for caste/biraderi membership and the presence of consanguinity could seriously jeopardize, and may totally invalidate, the results of association/case control studies and clinical trials. PMID- 19565014 TI - Molecular mechanisms of autosomal dominant and recessive distal renal tubular acidosis caused by SLC4A1 (AE1) mutations. AB - Mutations of SLC4A1 (AE1) encoding the kidney anion (Cl(-)/HCO(3) (-)) exchanger 1 (kAE1 or band 3) can result in either autosomal dominant (AD) or autosomal recessive (AR) distal renal tubular acidosis (dRTA). The molecular mechanisms associated with SLC4A1 mutations resulting in these different modes of inheritance are now being unveiled using transfected cell systems. The dominant mutants kAE1 R589H, R901X and S613F, which have normal or insignificant changes in anion transport function, exhibit intracellular retention with endoplasmic reticulum (ER) localization in cultured non-polarized and polarized cells, while the dominant mutants kAE1 R901X and G609R are mis-targeted to apical membrane in addition to the basolateral membrane in cultured polarized cells. A dominant negative effect is likely responsible for the dominant disease because heterodimers of kAE1 mutants and the wild-type protein are intracellularly retained. The recessive mutants kAE1 G701D and S773P however exhibit distinct trafficking defects. The kAE1 G701D mutant is retained in the Golgi apparatus, while the misfolded kAE1 S773P, which is impaired in ER exit and is degraded by proteosome, can only partially be delivered to the basolateral membrane of the polarized cells. In contrast to the dominant mutant kAE1, heterodimers of the recessive mutant kAE1 and wild-type kAE1 are able to traffic to the plasma membrane. The wild-type kAE1 thus exhibits a 'dominant-positive effect' relative to the recessive mutant kAE1 because it can rescue the mutant proteins from intracellular retention to be expressed at the cell surface. Consequently, homozygous or compound heterozygous recessive mutations are required for presentation of the disease phenotype. Future work using animal models of dRTA will provide additional insight into the pathophysiology of this disease. PMID- 19565016 TI - H9N2 avian influenza virus in a Mediterranean gull. PMID- 19565015 TI - Human Y-chromosome variation and male dysfunction. AB - The Y-chromosome is responsible for sex determination in mammals, which is triggered by the expression of the SRY gene, a testis-determining factor. This particular gene, as well as other genes related to male fertility, are located in the non-recombining portion of the Y (NRY), a specific region that encompasses 95% of the human Y-chromosome. The other 5% is composed of the pseudo-autosomal regions (PARs) at the tips of Yp and Yq, a X-chromosome homologous region used during male meiosis for the correct pairing of sexual chromosomes. Despite of the large size of the human NRY (about 60 Mb), only a few active genes are found in this region, most of which are related to fertility. Recently, several male fertility dysfunctions were associated to microdeletions by STS mapping. Now that the complete genetic map of the human Y-chromosome is available, the role of particular NRY genes in fertility dysfunctions is being investigated. Besides, along with the description of several nucleotide and structural variations in the Y-chromosome, the association between phenotype and genotype is being addressed more precisely. Particularly, several research groups are investigating the association between Y-chromosome types and susceptibility to certain male dysfunctions in different population backgrounds. New insights on the role of the Y-chromosome and maleness are being envisaged by this approach. PMID- 19565017 TI - Ab initio base fragment molecular orbital studies of influenza viral hemagglutinin HA1 full-domains in complex with sialoside receptors. AB - Mutations in avian influenza A viral hemagglutinin HA1 domain may alter the binding specificity of HA for alpha-sialosaccharide receptors, shifting the virus's host range from birds to humans. The amino acid mutations can occur at the sialoside binding site, as well as the antigenic site, far from the binding site. Thus, a theoretical study involving the in silico prediction of HA sialosaccharide binding may require quantum chemical analysis of HA1 full domain complexed with sialosides, balancing a computational cost with model size of HA1 sialoside complex. In addition, there is no insight to relationship between the model size of HA1-sialoside complex and its binding energy. In this study, H3 subtype HA1 full domains complexed with avian- and human-type Neu5Acalpha(2-3 and 2-6)Gal receptor analogs was investigated by ab initio based fragment molecular orbital (FMO) method at the level of second-order Moller-Plesset perturbation (MP2)/6-31G. Using this approach, we found avian H3 HA1 to bind to avian alpha2-3 receptor more strongly than to human alpha2-6 receptor in gas phase, by a value of 15.3-16.5 kcal/mol. This binding benefit was larger than that in the small model complex. Analysis of the interfragment interaction energies (IFIEs) between Neu5Ac-Gal receptor and amino acid residues on the full domain of H3 HA1 also confirmed the higher avian H3-avian alpha2-3 binding specificity. It was particularly important to evaluate the IFIEs of amino acid residues in a 13A radius around Neu5Ac-Gal to take account of long-range electrostatic interactions in the larger HA1-sialoside complex model. These results suggest suitable size of HA1-sialoside complex is significant to estimate HA1-sialoside binding energy and IFIE analysis with FMO method. PMID- 19565018 TI - The pig as a mixing vessel for influenza viruses: Human and veterinary implications. AB - Influenza A viruses are highly infectious respiratory pathogens that can infect many species. Birds are the reservoir for all known influenza A subtypes; and novel influenza viruses can emerge from birds and infect mammalian species including humans. Because swine are susceptible to infection with both avian and human influenza viruses, novel reassortant influenza viruses can be generated in this mammalian species by reassortment of influenza viral segments leading to the "mixing vessel" theory. There is no direct evidence that the reassortment events culminating in the 1918, 1957 or 1968 pandemic influenza viruses originated from pigs. Genetic reassortment among avian, human and/or swine influenza virus gene segments has occurred in pigs and some novel reassortant swine viruses have been transmitted to humans. Notably, novel reassortant H2N3 influenza viruses isolated from the US pigs, most likely infected with avian influenza viruses through surface water collected in ponds for cleaning barns and watering animals, had a similar genetic make-up to early isolates (1957) of the H2N2 human pandemic. These novel H2N3 swine viruses were able to cause disease in swine and mice and were infectious and highly transmissible in swine and ferrets without prior adaptation. The preceding example shows that pigs could transmit novel viruses from an avian reservoir to other mammalian species. Importantly, H2 viruses pose a substantial risk to humans because they have been absent from mammalian species since 1968 and people born after 1968 have little preexisting immunity to the H2 subtype. It is difficult to predict which virus will cause the next human pandemic and when that pandemic might begin. Importantly, the establishment and spread of a reassorted mammalian-adapted virus from pigs to humans could happen anywhere in the world. Therefore, both human and veterinary research needs to give more attention to potential cross-species transmission capacity of influenza A viruses. PMID- 19565019 TI - Host envelope glycoprotein processing proteases are indispensable for entry into human cells by seasonal and highly pathogenic avian influenza viruses. AB - Influenza A virus (IAV) is one of the most common infectious pathogens in humans and causes considerable morbidity and mortality. The recent spread of highly pathogenic avian IAV H5N1 viruses has reinforced the importance of pandemic preparedness. In the pathogenesis of IAV infection, cellular proteases play critical roles in the process of viral entry into cells that subsequently leads to tissue damage in the infected organs. Since there are no processing protease for the viral membrane fusion glycoprotein hemagglutinin precursor (HA(0)) in IAV, entry of the virus into cells is determined primarily by the host cellular HA(0) processing proteases that proteolytically activate membrane fusion activity. HA(0) of seasonal human IAV has the consensus cleavage site motif Q(E) T/X-R and is selectively processed by at least seven different trypsin-type processing proteases identified to-date in animal model experiments using mouse adapted IAV or gene expression system in MDCK cells. As is the case for the highly pathogenic avian influenza (HPAI) A virus, endoproteolytic processing of the HA(0) occurs through ubiquitous cellular processing proteases, which selectively recognize the multi-basic consensus cleavage site motifs, such as R-X K/R-R, and K-X-K/R-R. The cleavage enzymes for the R-X-K/R-R motif, but not K-X K/R-R motif, have been reported to be furin and pro-protein convertase (PC)5/6 in the trans-Golgi network. Here we report new members of type II transmembrane serine proteases of the cell membrane, mosaic serine protease large form (MSPL) and its splice variant TMPRSS13, which recognize and cleave both R-X-K/R-R and K X-K/R-R motifs without calcium. Furthermore, IAV infection significantly up regulates a latent ectopic pancreatic trypsin, one of the potent HA processing proteases, and pro-matrix metalloprotease-9, in various organs. These proteases may synergistically damage the blood-brain barrier in the brain and basement membrane of blood vessels in various organs, resulting in severe edema and multiple organ failure. In this review, we discuss these proteases as new drug target molecules for IAV treatment acting by inhibition of IAV multiplication and prevention of multiple organ failure, other than anti-viral agents, viral neuraminidase inhibitors. PMID- 19565020 TI - Controlling avian influenza infections: The challenge of the backyard poultry. PMID- 19565021 TI - Influenza virus pathogenicity is determined by caspase cleavage motifs located in the viral proteins. AB - Almost all influenza virus proteins are found to contain caspase cleavage motifs. Two caspase cleavage consensus sequences, EXD downward arrowY and D/EXXD downward arrowY (caspase motifs) were identified in N- and C-terminal regions of influenza virus proteins nucleocapsid NP (positions D(16) and D(497)) and ionic channel M2 (positions D(23) and D(87)). Using reverse genetics with the highly-virulent avian influenza virus A/FPV/Rostock (H7N1), as a vector precursor, these NP and M2 caspase motifs were artificially altered by site-directed mutagenesis and pathogenicity of the generated caspase mutant viruses was tested in chickens. Three main groups of virus mutants were identified. The first group of mutants was characterized by high replication in cells and low virulence in chickens. These virus mutants possessed the altered N-terminal NP and C-terminal M2 caspase motifs. The second group of virus mutants, possessing the altered N-terminal caspase motif of M2, was characterized by attenuated replication in cultured cells and reduced pathogenic properties in chickens. Third, mutations generated in the C-terminus of NP were lethal and restricted virus rescue by reverse genetics, implying a critical role of this caspase site in virus replication. Thus, these data suggested that, (i) caspase motifs in virus proteins play a significant role in virus pathogenicity; (ii) the lack of direct correlation between replication potential and pathogenicity, observed in caspase mutants of the first virus group, implied that virus caspase motifs could affect immunopathogenesis during the infection process, rather than simply controlling virus production in target cells in the chicken host. PMID- 19565022 TI - Differences in influenza virus receptors in chickens and ducks: Implications for interspecies transmission. AB - Avian influenza viruses are considered to be key contributors to the emergence of human influenza pandemics. A major determinant of infection is the presence of virus receptors on susceptible cells to which the viral haemagglutinin is able to bind. Avian viruses preferentially bind to sialic acid alpha2,3-galactose (SAalpha2,3-Gal) linked receptors, whereas human strains bind to sialic acid alpha2,6-galactose (SAalpha2,6-Gal) linked receptors. While ducks are the major reservoir for influenza viruses, they are typically resistant to the effects of viral infection, in contrast to the frequently severe disease observed in chickens. In order to understand whether differences in receptors might contribute to this observation, we studied the distribution of influenza receptors in organs of ducks and chickens using lectin histochemistry with linkage specific lectins and receptor binding assays with swine and avian influenza viruses. Although the intestinal epithelial cells of both species expressed only SAalpha2,3-Gal receptors, we found widespread presence of both SAalpha2,6-Gal and SAalpha2,3-Gal receptors in many organs of both chickens and ducks. Co-expression of both receptors may allow infection of cells with both avian and human viruses and so present a route to genetic reassortment. There was a marked difference in the primary receptor type in the trachea of chickens and ducks. In chicken trachea, SAalpha2,6-Gal was the dominant receptor type whereas in ducks SAalpha2,3-Gal receptors were most abundant. This suggests that chickens could be more important as an intermediate host for the generation of influenza viruses with increased ability to bind to SAalpha2,6-Gal receptors and thus greater potential for infection of humans. Chicken tracheal and intestinal epithelial cells also expressed a broader range of SAalpha2,3-Gal receptors (both beta(1-4)GlcNAc and beta(1-3)GalNAc subtypes) in contrast to ducks, which suggests that they may be able to support infection with a broader range of avian influenza viruses. PMID- 19565023 TI - Molecular quantitation of H9N2 avian influenza virus in various organs of broiler chickens using TaqMan real time PCR. AB - During the past decade, H9N2 low pathogenic avian influenza virus (LPAI) has caused considerable economic loss due to decreased production, increased mortality and the cost of vaccination in Iranian poultry industry. Because of widespread occurrence of this disease and the virus potential to mutate to highly pathogenic (HP) form and transmission to humans, it is, therefore, imperative to understand the pathogenesis and properties of these viruses. In this study, a two step TaqMan real time PCR assay was performed for the quantitation of A/chicken/Iran/772/1998(H(9)N(2)) virus in various organs of broiler chickens at different days post inoculation (DPI). Forty 5-week-old commercial broiler chickens were inoculated with the virus. Five chickens were randomly selected on days 1, 3, 6 and 9 PI. Their trachea, lungs, spleen, kidneys, pancreas, blood and faeces were collected for virus detection. A PCR test was performed and the positive samples were used for quantitative real time PCR assay. The result of RT PCR assay showed the presence of the virus in trachea (40%, 33%), lungs (20%, 66.6%) and spleen (20%, 50%) of infected chickens on days 3 and 6 PI, respectively. The virus was also detected in the kidneys of inoculated chickens on 3 (40%), 6 (60%) and 9 (100%) DPI. In faecal samples the virus was only detected on day 6 PI (83.3%). The molecular quantitation of AIV showed that the AIV titre in the trachea, lungs and spleen of chickens at 3 DPI is lower than the AIV titre at 6 DPI in these organs. The highest titre was observed in the faeces. The AIV titre in all organs of the birds which died at 6 DPI was higher than those of the same organs in the other experimental birds. PMID- 19565024 TI - The Stewart approach--one clinician's perspective. AB - Peter Stewart added controversy to an already troubled subject when he entered the clinical acid-base arena. His approach puts water dissociation at the centre of the acid-base status of body fluids. It is based on six simultaneous equations, incorporating the Laws of Mass Action, Mass Conservation, and Electrical Neutrality. Together with Gibbs-Donnan equilibria, these equations explain the diagnostically important PaCO(2)/pH relationship, and improve understanding of the physiologic basis of traditional acid-base approaches. Spin offs have included new scanning tools for unmeasured ions, in particular the 'strong ion gap' and 'net unmeasured ions'. The most controversial feature is the designation of pH and bicarbonate concentrations as dependent variables, answerable exclusively to three independent variables. These are the strong ion difference (SID), the total concentration of non-volatile weak acid (A(TOT)), and PCO(2). Aspects of this assertion conflict with traditional renal physiology, and with current models of membrane H(+)/base transporters, oxidative phosphorylation, and proton and bicarbonate ionophores. The debate in this area is ongoing. Meanwhile, Stewart-style diagnostic and decision support systems such as the 'Strong Ion Calculator' and the web-site www.acidbase.org are now appearing. PMID- 19565025 TI - Clinical use of pharmacogenomic tests in 2009. AB - Pharmacogenomics is a new field where testing an individual can define either a risk status for an adverse event, or the rate of metabolism of a drug. This is achieved by the categorisation of the enzyme activity or documenting genetic polymorphisms of a metabolising enzyme. The best example of risk status assessment is the recent finding that HLA-B typing a person can predict whether they are at risk of a severe skin reaction from the drug abacavir. Those patients showing HLA-B*5701, who are being considered for abacavir therapy, can be prevented from developing potentially toxic epidermal necrosis (TEN) or Stevens Johnson Syndrome by avoiding abacavir. The evidence for HLA-B typing for allopurinol and carbamazepine has also been described. Most other pharmacogenomic tests are of drug metabolising enzymes, which can either be assessed using "probe" drugs and measuring a ratio of parent drug to metabolite, or, by genetic testing for polymorphisms of the genes. In practice, testing is usually done by molecular testing, but this typically does not detect all polymorphisms. This article briefly reviews the evidence for the utilisation of pharmacogenomics for antidepressant drugs, tamoxifen, codeine, warfarin, azathioprine, clopidogrel, omeprazole, tacrolimus and irinotecan. There are few pharmacogenomics tests being carried out in practice, as there has not been a wide appreciation of their use, and only limited evidence exists for many individual drugs. It is expected that utilisation will increase as more evidence becomes available and there is a wider understanding of the existing evidence by the medical profession. PMID- 19565026 TI - Investigating maturity onset diabetes of the young. AB - Maturity Onset Diabetes of Young (MODY) is a monogenic and autosomal dominant form of diabetes mellitus with onset of the disease often before 25 years of age. It is due to dysfunction of pancreatic beta cells characterised by non-ketotic diabetes and absence of pancreatic auto-antibodies. It is frequently mistaken for type 1 or type 2 diabetes mellitus. Diagnosis of MODY is important as the GCK subtype has better prognosis and may not require any treatment. Subtypes HNF1A and HNF4A are sensitive to sulfonylureas, however diabetes complications are common if not treated early. Moreover, there is genetic implication for the patient and family. Rare MODY subtypes can be associated with pancreatic and renal anomalies as well as exocrine dysfunction of the pancreas. So far there are six widely accepted subtypes of MODY described but the list has grown to nine. Although the majority of diabetes mellitus in youth remains type 1 and the incidence of type 2 is rising, MODY should be considered in patients with non ketotic diabetes at presentation, and in patients with a strong family history of diabetes mellitus without pancreatic auto-antibodies. Furthermore the diagnosis must be confirmed by molecular studies. With advancement in genomic technology, rapid screening for MODY mutations will become readily available in the future. PMID- 19565027 TI - The clinical and biochemical spectrum of congenital adrenal hyperplasia secondary to 21-hydroxylase deficiency. AB - 21-Hydroxylase Deficiency (21-OH Deficiency) represents the most common form of Congenital Adrenal Hyperplasia (CAH), a complex and heterogenous group of conditions, characterised by defects in one of the five enzymes involved in adrenal steroidogenesis. Defects in this steroidogenic enzyme, the product of the CYP21A2 gene, cause disruption in the pathway involved in cortisol and aldosterone production and consequently, the accumulation of their steroid precursors as well as a resulting adrenocorticotrophic hormone (ACTH)-driven overproduction of adrenal androgens. Treatment with glucocorticoid, with or without mineralocorticoid and salt replacement, is directed at preventing adrenal crises and ensuring normal childhood growth by alleviating hyperandrogenism. Conventionally, two clinical forms of 21-OH Deficiency are described - the classical form, separated into salt-wasting and simple-virilising phenotypes, and the non-classical form. They are differentiated by their hormonal profile, predominant clinical features and age of presentation. A greater understanding of the genotype-phenotype correlation supports the view that 21-OH Deficiency is a continuum of phenotypes as opposed to a number of distinct phenotypical entities. Significant advancements in technologies such as Tandem Mass Spectrometry (TMS) and improvements in gene analysis, such as complete PCR-based sequencing of the involved gene, have resulted in remarkable developments in the areas of diagnosis, treatment and treatment monitoring, neonatal screening, prenatal diagnosis and prenatal therapy. PMID- 19565028 TI - Animal models of cancer in the head and neck region. AB - Animal models that resemble the cancers of the head and neck region are of paramount importance in studying the carcinogenesis of these diseases. Although several methods for modeling cancer in the head and neck are available, none are fully satisfactory. Subcutaneous xenograft models of cancer in nude mice are often used in preclinical studies. However, these models are problematic in several aspects as they lack the specific interactions that exist between the tumor cells and their native environment. Establishment of tumors at the orthotopic sites restore these distinct patterns of interactions between the tumor and the host organs that are lost or altered when the tumors are established in ectopic sites. With regard to the transgenic model of cancer in the head and neck region, it should be kept in mind that the transgene used to drive the malignant transformation may not be representative of the carcinogenic process found in human tumors. Low penetrance of tumor formation also translates into high cost and time commitment in performing studies with transgenic models. In this review, we will discuss some of the commonly used methods for modeling cancer in the head and neck region including squamous cell carcinoma of the head and neck as well as thyroid carcinoma. PMID- 19565029 TI - Pre-operative Evaluation of Eustachian Tube Function Using a Modified Pressure Equilibration Test is Predictive of Good Postoperative Hearing and Middle Ear Aeration in Type 1 Tympanoplasty Patients. AB - OBJECTIVES: The Eustachian tube (ET) plays an important role in maintaining a normally aerated middle ear. Inflammation in middle ear disease is related to ET dysfunction, and postoperative restoration of middle ear integrity and hearing are closely related to ET function in chronic ear disease patients. After successful tympanoplasty, restoration of a well-aerated middle ear with good ET function can permit better compliance of the tympanic membrane. In this study, we evaluated the predictive validity of preoperative ET function measurements. METHODS: We reviewed 137 patients who underwent type 1 tympanoplasty. All patients had non-cholesteatomatous chronic otitis media and received canal wall up-type tympanomastoidectomies. Patients were categorized into four groups according to preoperative ET function measurements using a modified pressure inflation-deflation equilibration test. Group I patients had residual pressures less than 10 daPa, and Group IV patients showed no pressure change (poor results). Groups II and III were intermediate. Hearing levels were determined using pure tone averages at four frequencies. Postoperative tympanography was performed to determine middle ear aeration. RESULTS: The preoperative air bone (AB) gap was 29.6+/-7.0 dB, and the postoperative gap was 16.5+/-5.7 dB; thus, there was significant overall improvement. In all groups, hearing was significantly better after surgery, but the worst postoperative hearing level was seen in Group IV patients. Type B tympanograms were more frequently recorded in Group IV patients than they were in Group I or II patients. Postoperative AB gaps were 9.2+/-3.8 dB in patients with type A tympanograms, 13.4+/-2.1 dB in those with type As, 24.1+/-2.5 dB in those with type C, and 18.5+/-2.8 dB in those with type B. CONCLUSION: ET function measured with a modified pressure equilibration test using an inflation-deflation manometric method is a good indicator of an aerated middle ear and is predictive of improved postoperative hearing. PMID- 19565030 TI - Granulocytic Sarcoma in the Head and Neck: CT and MR Imaging Findings. AB - OBJECTIVES: To evaluate characteristic computed tomography (CT) and magnetic resonance (MR) imaging findings of granulocytic sarcomas of the head and neck. METHODS: The CT (n=11) and MR (n=1) images obtained from 11 patients (7 males and 4 females; mean age, 23.5 yr; age range, 1 to 69 yr) with histologically-proven granulocytic sarcomas of the head and neck were retrospectively reviewed. Histological confirmation was done by bone marrow biopsy in 9 patients, and/or local biopsy in 4 patients. The imaging findings were analyzed with particular attention to location, size, shape, margin, bone destruction, internal architecture, pattern and degree of enhancement, and multiplicity of the lesions. RESULTS: The masses were most commonly located in the orbital cavity (n=8); other locations included lymph nodes (n=5) and palatine/pharyngeal/lingual tonsils (n=3). The mass sizes varied from a mean diameter of 1.3 to 5.8 cm (average, 2.6 cm). Multiple lesions were found in 6 patients. The shapes of the tumors were ovoid in 12 patients and irregular in 4 patients. Most lesions had poorly-defined margins (13/16) and invaded adjacent bony structures (5/16). On the pre-contrast CT images, the masses were iso- (5/8) or low-density (3/8) in comparison with muscle. The MRI, which was obtained in one patient in this study, showed that the mass was iso-signal intensity on T1-weighted images and iso-signal intensity on T2-weighted images compared to the gray matter of the brain. On the post-contrast CT images, there was homogenesous (n=12) or heterogeneous (n=4) enhancement, with mild (n=10), moderate (n=4), and marked (n=2) enhancement in the solid portions of the lesions. CONCLUSION: Although rare, granulocytic sarcomas arise in various locations in the head and neck area (most commonly in the orbit) in the form of well-demarcated, and mildly- and homogenously-enhancing masses with adjacent bony invasion. PMID- 19565031 TI - Patterns of Post-thyroidectomy Hemorrhage. AB - OBJECTIVES: Postoperative hemorrhage is a potentially life-threatening complication in thyroid surgery. This study was performed to review the clinical patterns of post-thyroidectomy hemorrhage, and especially as they are related to the source of bleeding. METHODS: We performed a retrospective review of 10 patients (0.96%) with post-thyroidectomy hemorrhage that required surgical evacuation. The clinical patterns such as the time interval from surgery to hemorrhage and the signs and symptoms according to the bleeding focus were evaluated. RESULTS: The mean time interval from surgery to symptom onset was 7 hr 52 min. Six cases showed bleeding deep to the strap muscles, while the other 4 cases showed bleeding superficial to the muscles. Ecchymosis was prominent and dark in color in 3 of the 4 cases (75%) of superficial bleeding, however it was identified in only 2 of the 6 cases (33%) of deep bleeding. Respiratory distress occurred in two cases of hematoma deep to the strap muscles, but in none of the cases with superficial bleeding. CONCLUSION: The post-thyroidectomy hemorrhage had some different clinical patterns between the superficial cases and the deep cases, showing that life-threatening airway obstruction occurred from the deep hematoma. A thorough understanding of the clinical patterns of post-thyroidectomy hemorrhage between the cases of superficial and those cases of deep hematoma may provide valuable surgical tips to manage this potentially lethal complication. PMID- 19565032 TI - Clinical efficacy of primary tumor volume measurements: comparison of different primary sites. AB - OBJECTIVES: The purpose of study was to determine the clinical efficacy of primary tumor volume measurements of different primary sites in the oropharynx compared to the oral cavity. METHODS: A retrospective analysis of 85 patients with oral cavity or oropharynx cancer. The tumor area was manually outlined from axial magnetic resonance (MR) series. The software calculated the tumor volumes, automatically. The values of the primary tumor volumes were then subdivided into separate groups (3,500 mm(3)). RESULTS: The prognostic indicators were the cT and cN (oral cavity); age, primary site, cT, cN, and primary tumor volume (oropharynx) on the univariate analysis. There was no significant prognostic factor for oral cavity cancer on the multivariate analysis. Primary site, cN, and primary tumor volume were independent prognostic indicators for oropharynx cancer by multivariate analysis. CONCLUSION: Primary tumor volume measurement is a reliable way to stratify outcome, and make up for the weak points in the American Joint Committee on Cancer staging system with oropharynx cancer. PMID- 19565033 TI - Comparison of Conventional Excision via a Sublabial Approach and Transnasal Marsupialization for the Treatment of Nasolabial Cysts: A Prospective Randomized Study. AB - OBJECTIVES: Surgical excision via a sublabial approach is considered the standard treatment for nasolabial cysts. Although transnasal marsupialization has been proposed as an alternative method, no prospective study has compared the effectiveness of these techniques. We thus compared the surgical procedure, operating time, postoperative pain, complications, and recurrence rate between the two surgical methods. METHODS: Twenty patients diagnosed with nasolabial cysts were allocated randomly into two groups according to the surgical technique. In the sublabial approach group, the cysts were excised completely using a sublabial approach, while in the transnasal marsupialization group, the cysts were marsupialized transnasally under the guidance of nasal endoscopes. The pure operating time was measured and postoperative pain was evaluated using a visual analog scale. Complications after the procedure were assessed and recurrence was determined according to the clinical symptoms and postoperative radiologic findings. RESULTS: The transnasal marsupialization group had significantly shorter operating times, less postoperative pain, lower complication rates, and shorter duration of side effects than the sublabial approach group. No recurrence occurred in either group after a 1-yr follow-up period. CONCLUSION: Although both methods are effective for treating nasolabial cysts, the transnasal marsupialization of nasolabial cysts has many benefits over the conventional sublabial approach. Therefore, we propose that transnasal marsupialization be the treatment of choice for nasolabial cysts. PMID- 19565034 TI - Compliance with positive airway pressure treatment for obstructive sleep apnea. AB - OBJECTIVES: Positive airway pressure (PAP) is considered a standard treatment for moderate-to-severe obstructive sleep apnea (OSA) patients. However, compliance with PAP treatment is suboptimal because of several types of discomfort experienced by patients. This study investigated compliance with PAP therapy, and affecting factors for such compliance, in OSA patients. METHODS: We performed a survey on 69 patients who engaged in PAP therapy between December 2006 and November 2007. After diagnostic polysomnography and manual titration, patients trialed PAP using the ResMed instrument and explored autoadjusting PAP (APAP), continuous PAP (CPAP), and flexible PAP (using expiratory pressure relief [EPR]) at least once every week for 1 month. Compliance measures were mean daily use (hr), percentage of days on which PAP was used, and percentage of days on which PAP was used for >4 hr. Data were obtained at night using the software Autoscan version 5.7(R) of the ResMed Inc. We obtained data on anthropometric (age, BMI, neck circumflex, Epworth sleepiness scale, Pittsburgh Sleep Quality Index, hypertension, alcohol intake), polysomnographic data (severity of apnea-hypopnea index [AHI], proportion of nonsupine sleep time, position dependence of sleep), PAP mode and AHI during PAP use for affecting factors. RESULTS: After 1 month, 41 of the 69 patients (59.4%) were pleased with PAP therapy and purchased instruments. Twenty-four patients (34.7%) used PAP for more than 3 months. The percentage of days on which PAP was used was statistically higher in patients with hypertension than in normotensive patients (P=0.003). There were negative correlations 1) between nonsupine position sleep time and percentage of days on which PAP was used (r=-0.424, P=0.039), and 2) between the AHI during PAP use and the percentage of days on which PAP was used for >4 hr (r=-0.443, P=0.030). There were no statistical differences between AHI, BMI, PAP pressure, or other measured parameters, on the one hand, and compliance, on the other. CONCLUSION: The affecting factors for PAP use were hypertension history, sleep posture (shorter nonsupine sleep time), and lower AHI during PAP use. PMID- 19565035 TI - CSF Otorrhea Resulting from Osteoradionecrosis of the Temporal Bone in a Patient with Recurrent Meningioma. AB - Osteoradionecrosis of the temporal bone is a very rare but potentially lethal complication of radiotherapy for head and neck or skull base tumors. Only two cases of osteoradionecrosis of the temporal bone complicating cerebrospinal fluid (CSF) otorrhea have been reported in the literature. This report describes a case of CSF otorrhea and osteoradionecrosis of the temporal bone in a patient with meningioma who was treated with tympanomastoid surgery and autologous fat obliteration in the mastoid. PMID- 19565036 TI - A Family of H723R Mutation for SLC26A4 Associated with Enlarged Vestibular Aqueduct Syndrome. AB - Recessive mutations of the SLC26A4 (PDS) gene on chromosome 7q31 can cause sensorineural deafness with goiter (Pendred syndrome, OMIM 274600) or NSRD with goiter (at the DFNB4 locus, OMIM 600791). H723R (2168A>G) is the most commonly reported SLC26A4 mutations in Korean and Japanese and known as founder mutation. We recently experienced one patient with enlarged vestibular aqueduct syndrome. The genetic study showed H723R homozygous in the proband and H723R heterozygous mutation in his family members. The identification of a disease-causing mutation can be used to establish a genotypic diagnosis and provide important information to both families and their physicians. PMID- 19565037 TI - Gastric choristoma of the oropharynx. AB - Heterotopic gastric mucosa tissue is also called gastric choristoma, and this type of lesion can be found anywhere in the alimentary tract. However, gastric choristoma in the pharynx is very rare; only 10 cases of pharyngeal gastric choristoma have been reported in the English medical literature. A 32-yr-old woman was referred to our institution for the evaluation of a large mass that originated from the posterior wall of the oropharynx. The mass did not cause any symptoms except for the occasional sensation of a foreign body. Gadolinium enhanced T1 weighted imaging showed a 5 cm-sized mass with central enhancement and hypointense portions, yet the radiological diagnosis was not clear. Transoral mass excision was performed with using electrocautery for making the diagnosis and for treating the mass. The microscopic analysis revealed gastric choristoma. PMID- 19565038 TI - Looking for new features to improve accuracy of EUS lymph node criteria. PMID- 19565039 TI - Jews and inflammatory bowel disease. PMID- 19565041 TI - Diphyllobothrium latum identified by capsule endoscopy--an unusual cause of iron deficiency anemia. PMID- 19565040 TI - Prophylaxis of hepatitis B virus recurrence after liver transplantation. PMID- 19565042 TI - Investigation of the SPINK1 N34S mutation in Romanian patients with alcoholic chronic pancreatitis. A clinical analysis based on the criteria of the M-ANNHEIM classification. AB - BACKGROUND AND AIMS: The N34S mutation in the serine protease inhibitor Kazal type I (SPINK1) gene has been associated with chronic pancreatitis. Clinical data about the phenotypic expression of alcoholic chronic pancreatitis with the N34S variant are limited. The prevalence of the N34S mutation in patients with chronic pancreatitis and healthy individuals from Eastern Europe is unknown. METHODS: We studied Romanian patients with chronic pancreatitis and investigated the clinical presentation in patients with N34S mutation. The SPINK1 N34S variant was analysed in 94 chronic pancreatitis patients and 96 healthy controls by an allele specific PCR method and a restriction fragment length polymorphism method. A meta-analysis was conducted with previous N34S association studies. The clinical course of alcoholic pancreatitis was evaluated according to the severity criteria of the M ANNHEIM classification system of chronic pancreatitis. RESULTS: A heterozygous N34S mutation was found in 1 of 96 healthy individuals (1%) and in 4 of 80 patients (5%) with alcoholic chronic pancreatitis. The meta-analysis confirmed the status of N34S as a risk factor for the development of alcoholic chronic pancreatitis (OR=5.3). However, the clinical course of the disease was similar in patients with and without N34S mutation. CONCLUSION: The N34S mutation is a weak risk factor for alcoholic chronic pancreatitis. PMID- 19565043 TI - Obesity and functional constipation; a community-based study in Iran. AB - BACKGROUND: Many factors have been linked to the occurrence of constipation, but few studies exist regarding the link between obesity and constipation. The aim of this study was to assess the association between body mass index (BMI) and functional constipation in the Iranian community. METHODS: From May 2006 to December 2007, a cross sectional study was conducted in the Tehran province and a total of 18,180 adult persons were drawn up randomly. One questionnaire was filled in two stages through interviews. In the first part, personal characteristics and 11 gastrointestinal symptoms were listed. Those who reported at least one of these 11 symptoms were referred for the second interview. The second part of the questionnaire consisted of questions about different gastrointestinal disorders based on the Rome III criteria including functional constipation. RESULTS: 459 adult persons were found to have functional constipation. The mean +/- SD of BMI was 26.5 +/- 4.7 and 60% of the patients had a BMI more than 25. Age and education were significantly associated factors with obesity, showing that older patients and less educated patients were more overweight and obese. Smoking, marital status and sex were not significantly associated with obesity but, up to 60% of low educated women who had functional constipation, had a BMI more than 25. CONCLUSIONS: Our study showed that about 60% of patients with functional constipation were overweight, which was more than the mean of our community. In addition there may be an association between higher BMI level and the low education level with constipation in Iranian women. PMID- 19565044 TI - Clinical features of ulcerative colitis and Crohn's disease in Turkey. AB - BACKGROUND AND AIMS: This study is aimed at determining the clinical and socio demographic characteristics of Turkish patients with inflammatory bowel disease who have been treated in a tertiary referral center. PATIENTS AND METHODS: The study was conducted between 1993 and 2007 at Ankara Yuksek Ihtisas Hospital, Inflammatory Bowel Disease Clinics. The clinical and epidemiologic data from this clinic were gathered and analysed. RESULTS: During the study period, 702 patients were identified as having inflammatory bowel disease; among these, 507 with ulcerative colitis (72.2%) and 195 with Crohn's disease (27.8%). The mean age at diagnosis was 46.2 years in ulcerative colitis and 40 years in Crohn's disease (p < 0.001). The male to female ratio was 1.2 for ulcerative colitis and 1.6 for Crohn's disease (p = 0.12). Of the patients who were primarily diagnosed with ulcerative colitis 16.8% had their diagnoses changed to Crohn's disease after the follow-up period. CONCLUSIONS: Peak age of onset for both Crohn's disease and ulcerative colitis were similar to other countries. A slight male predominance of Crohn's disease and ulcerative colitis were found for Turkish patients. Though it takes longer to diagnose, Crohn's disease may have a milder course in Turkish patients. PMID- 19565045 TI - Activated liver stellate cells in chronic viral C hepatitis: histopathological and immunohistochemical study. AB - BACKGROUND: There is positive correlation between the number of activated hepatic stellate cells and necroinflammatory activity and/or the stage of liver fibrosis in viral hepatitis. No study has investigated such a relationship with regard to the activated hepatic cells within specified zones of liver tissue in chronic C hepatitis. The aim of the present study was to correlate the level of activated hepatic stellate cells within perivenular, intermediate, periportal, and portal tracts area and fibrous septa with stages of liver fibrosis and necroinflammatory activity in patients with chronic C hepatitis. METHODS: This retrospective study included 20 liver biopsy samples from patients with chronic C hepatitis and 10 normal liver biopsies. Biopsy specimens were processed routinely and stained with haematoxylin-eosin, periodic acid-Schiff, Masson;s trichrome, aldechide fuchsin, reticulin and iron (Pearls). Activated hepatic stellate cells were identified immunohistochemically using antibody to alpha-smooth muscle actin. Assessment of immunoreactivity was performed using a semiquantitative method. RESULTS: In chronic C hepatitis, a positive correlation between the stage of fibrosis and the number of activated hepatic stellate cells within portal spaces and fibrous septa was found. These cells were increased in number in other areas of liver tissue as well, but without statistical significance. There was no correlation between either the stage of fibrosis and necroinflammatory activity or the number of activated hepatic stellate cells and necroinflammatory activity. CONCLUSION: An increased number of activated hepatic stellate cells within portal spaces and fibrous septa may be a useful prognostic marker for the development of advanced fibrosis and cirrhosis in chronic C hepatitis. PMID- 19565046 TI - Histopathologic outcome of neoadjuvant image-guided therapy of hepatocellular carcinoma. AB - BACKGROUND AND AIMS: The best curative treatment for hepatocellular carcinoma (HCC) is liver transplant (LT), with the limitation to either a solitary lesion < 5 cm or up to three lesions < 3 cm each. Arresting tumor growth or downstaging to make patients eligible for LT can be obtained by neoadjuvant treatments such as transarterial chemoembolization (TACE), selective internal radiation therapy (SIRT), chemical or radiofrequency ablation (RFA). We evaluated the histopathologic response in explant specimens to neoadjuvant image-guided therapy of HCC prior to LT. METHODS: Twenty-eight patients with 39 HCC nodules eligible for LT underwent neoadjuvant image-guided therapy 1-393 days prior to transplant. Treatment included TACE (5 nodules), SIRT (7 nodules), RFA (12 nodules), chemical ablation (3 nodules) combined TACE and acetic acid injection (1 nodule) and combined TACE and RFA (11 nodules). 19/28 patients not transplanted within 30 days had interval MRI and 3 patients with progressive disease were retreated. RESULTS: Residual viable tumor was seen in 42% of patients with post-treatment imaging. Explant pathology revealed viable tumor in 35 of 39 (90%) treated nodules and somewhere in the explanted liver in all patients. Viability and/or progression of the treated tumor was noted in 5/5 nodules treated with TACE, 6/7 with SIRT, 11/12 with RFA, 2/3 with chemical ablation and 11/12 with combined treatment. CONCLUSION: Viable local or remote tumor was identified on explanted liver in the majority of patients with HCC after neoadjuvant therapy, despite apparent successful treatment on MRI. PMID- 19565047 TI - Endosonographic surveillance of small gastrointestinal tumors originating from muscularis propria. AB - INTRODUCTION: Submucosal tumors (SMT) are not uncommonly found during upper endoscopy. Management for small SMT originating from muscularis propria (MP) is controversial. Data regarding regular endoscopic ultrasound (EUS) surveillance is scarce. We report our experience in using EUS to monitor these tumors. METHODS: Patients with SMT originated from MP as confirmed by EUS were recruited. The maximal diameter, echo pattern, presence of cystic spaces and regularity of extra luminal margin were documented. Patients with large tumors (maximal diameter > 3 cm), heterogeneous echo pattern, presence of cystic spaces or irregular extra luminal margin were offered surgery in view of malignant risk. Patients with small tumor and benign EUS features were offered regular EUS surveillance or surgery if they wished. The progress of those patients who underwent EUS surveillance were studied. RESULTS: From January 2002 to December 2007, there were 93 patients with SMT originating from MP. Forty-nine patients had a small tumor and benign EUS features. Only two of these patients chose surgery. The histopathological results were low risk gastrointestinal stromal tumor (GIST) in both patients. Twenty-three patients elected to undergo regular EUS surveillance for a mean period of 17.3 months (range 6-42 months). Three patients (13.0%) showed interval increase in tumor size. There was no change in other EUS features. Surgery was performed in these 3 patients. Histological examination revealed schwannoma in 2 patients and low risk GIST in 1 patient. CONCLUSION: It remains unclear whether EUS surveillance for small tumors originating from MP in the upper gastrointestinal tract is useful. PMID- 19565048 TI - Irregular echogenic foci representing coagulation necrosis: a useful but perhaps under-recognized EUS echo feature of malignant lymph node invasion. AB - BACKGROUND AND AIMS: Coagulation necrosis has been described in malignant lymph nodes. Our aim was to determine if coagulation necrosis in mediastinal lymph nodes imaged by EUS could be used as a useful echo feature for predicting malignant invasion. METHODS AND DESIGN: Patients with known or suspected lung cancer who had undergone mediastinal lymph node staging by EUS. SETTING: Tertiary Care university hospital. PARTICIPANTS AND INTERVENTION: An expert endosonographer blinded to the final diagnosis, reviewed the archived digital EUS images of lymph nodes prior to being sampled by FNA. LNs positive for malignancy by FNA were included. The benign group included lymph node images with either negative EUS-FNA or lymph nodes imaged by EUS but not subjected to EUS-FNA, with surgical correlation of their benign nature. RESULTS: 24 patients were included. 8 patients were found to have coagulation necrosis. 7/8 patients had positive result for malignancy by EUS-FNA. One patient determined to have coagulation necrosis had a non-malignant diagnosis indicating a false positive result. 16 patients had no coagulation necrosis. In 6 patients with no coagulation necrosis, the final diagnosis was malignant and in the remaining 10 cases, the final diagnosis was benign. For coagulation necrosis as an echo feature for malignant invasion, sensitivity was 54%, specificity was 91%, positive predictive value was 88%, negative predictive value was 63% and accuracy was 71%. CONCLUSION: Coagulation necrosis is a useful echo feature for mediastinal lymph node staging by EUS. PMID- 19565049 TI - Evaluation of guide wire cannulation technique in elderly patients with choledocholithiasis. AB - AIMS: Post-ERCP pancreatitis is the most frequent complication of ERCP. We aimed to evaluate guide-wire cannulation as compared to conventional contrast-assisted cannulation with regard to the rate of post-ERCP pancreatitis (PEP) in elderly patients with choledocholithiasis. METHODS: Patients aged 80 years or over with known choledocholithiasis, who were referred to ERCP in one single district hospital from January 2005 to March 2008 were reviewed retrospectively. RESULTS: During this period, 246 ERCPs were performed in 217 patients. In 82 procedures (37.0%) deep cannulation of the biliary tree was achieved by means of a contrast assisted procedure, whereas in the remaining 135 attempts a guide wire was used. Among the 64 patients older than 80 years, cannulation was accomplished in 25 by contrast visualization of the biliary tree and in 39 by means of the guide-wire. In the overall population of 217 patients, PEP occurred in 11 patients (5%), 10 and 1 case, respectively, for each of the two procedures (p = 0.00042). In the elderly patients, PEP occurred in 5 out of 25 patients after contrast filling of the bile duct, and in 1 out of 39 patients after the guide-wire utilization (p = 0.021). Cannulation was successful in 201 of 217 patients (92.6%), and in 57 of the 64 elderly patients (89%) (36 with guide wire, p = 0.42). Bleeding occurred in 5 patients (2.3%) and perforation in one (0.46%). CONCLUSION: The guide-wire seems to reduce the incidence of pancreatitis in the elderly compared to conventional contrast but does not improve the success rate for cannulation. PMID- 19565050 TI - Staging laparoscopy in gastric cancer. Accuracy and impact on therapy. AB - BACKGROUND: Surgical therapy remains the most effective treatment modality in gastric cancer. The importance of multimodal treatment for advanced gastric cancer has contributed to the development of more accurate preoperative staging strategies. We examined the diagnostic accuracy of staging laparoscopy (SL) for abdominal metastases and the predictive value of SL for tumor resectability. MATERIAL AND METHOD: This is a prospective, cohort, observational study of 98 patients with primary gastric adenocarcinoma admitted at a tertiary referral hospital over a three year period. Extended SL, laparoscopic ultrasonography and peritoneal cytology were performed in 45 patients with gastric cancer without distant metastases on pre-therapeutic imaging staging. Of the 45 patients, 17 (37.8%) had distant metastases on SL and were offered palliative therapy and/or supportive care. Open laparotomy and gastrectomy was performed in the patients without distant metastases or with uncertain resectability on SL. RESULTS: An unnecessary laparotomy was avoided in 17 (37.8%) patients. The overall SL sensitivity for distant metastases was 89%, specificity 100% and diagnostic accuracy 95.5%. The sensitivity for lymph node metastases was 54.5%, the specificity 100% and the diagnostic accuracy 64.3%. The SL positive predictive value for resectability was 96% and the negative predictive value was 50%. The morbidity of SL was 2.2% and the mortality 0. CONCLUSION: Staging laparoscopy is a safe and effective staging modality in patients with gastric carcinoma. It avoids unnecessary laparotomies in a significant number of patients and should be mandatory if neoadjuvant treatment is planned. PMID- 19565051 TI - Prognostic factors of liver injury in polytraumatic patients. Results from 895 severe abdominal trauma cases. AB - BACKGROUND AND AIMS: Prognosis of multiple injured patients is mainly limited by severe haemorrhage. Although mechanisms of altered immune response have been intensively investigated, little is known about the relevance of liver trauma as an independent predictive outcome factor in these patients. METHODS: 10,469 patients from the DGU Trauma Registry (1993-2005) were retrospectively analyzed. Primary admitted patients with an injury severity score > or = 16, without isolated head injury were included. Patients were analyzed according to the injury pattern as liver injury (Abbreviated Injury Scale--AIS abdomen < 3 and AIS liver 2-5; n = 321), non-liver abdominal trauma (AIS abdomen 2-5 or AIS liver < 3; n = 574) and control group without abdominal injuries (AIS abdomen or liver < 3; n = 9,574). RESULTS: Severe liver injury was associated with excessive demands for volume resuscitation and induced a significantly increased risk for sepsis and multi-organ failure (MOF) compared to both other groups (sepsis 19.9% vs. 11.0%; MOF 32.7% vs. 16.6%). Furthermore, deleterious outcome was more frequently associated with severe liver trauma (mortality 34.9%) compared to severe abdominal trauma (12.0%). CONCLUSION: Severe liver trauma is an independent predictor for severe haemorrhage with a substantially increased risk of sepsis, MOF and trauma-related death. While conservative treatment of patients with liver trauma but no haemorrhage is effective, patients with hemodynamic instability seem to be from a subgroup where contemporary treatment modalities are not yet sufficient. PMID- 19565052 TI - Assessing gastrointestinal symptoms and perception, quality of life, motility, and autonomic neuropathy in clinical studies. AB - Imprecise characterization of complaints of the upper and lower gastrointestinal (GI) tract puts patients at risks of either a delayed diagnosis or misdiagnosis and contributes to an increase in the overall direct and indirect costs of the health system. The current scenario in the case of functional GI diseases originates from at least two conditions: frequency of diseases and bothersome symptoms with an impact on the quality of life (QoL). To make a correct diagnosis is therefore almost mandatory. Once a positive diagnosis of functional involvement of the GI tract is made, the correct diagnosis assessment includes the study of symptom characteristics, entity and perception, detection of abnormal patterns of GI motor-function (gallblader and gastric emptying, oro cecal and colonic transit, etc.), potential involvement of the autonomic nervous system (sympathetic, parasympathetic), and overall impact of such abnormalities on the QoL and psychological profiles. Results of these tests can be variable, depending on the type and intensity of the illness. In the present review, the state-of-the-art methods for correct assessment of several factors regarding the onset, perpetuation and outcome of functional GI diseases are discussed. PMID- 19565053 TI - An interesting ca(u)se of small bowel obstruction. PMID- 19565054 TI - Electroincision of anastomotic stricture after curative resection for gastric cancer. PMID- 19565055 TI - Mitochondrial alterations in nonalcoholic fatty liver disease. Pediatric case description of three submitted sequential biopsies. AB - Nonalcoholic fatty liver disease (NAFLD) is a clinical-pathological syndrome that encompasses a wide spectrum of morphologic alterations, ranging from simple hepatic steatosis to a more severe stage, known as nonalcoholic steatohepatitis (NASH). The purpose of this clinical report was to contribute to the understanding of mitochondrial alterations in NAFLD. The child (13-month-old) underwent initial biopsy in the year 2000 and was diagnosed with diffuse macro and microvesicular steatosis. Two additional biopsies were performed in 2001 and 2004. A high percentage of microvesicular steatosis was observed in the biopsies performed in 2000 and 2001. Mitochondrial size was slightly increased in the biopsy performed in the year 2000, significantly increased in 2001 and decreased in 2004. The presence of "mitochondrial hypertrophy" in the hepatocytes of an asymptomatic pediatric patient whose disease presentation was typical of NAFLD, excluding other pathological processes, allowed us to suspect that such a defect was considered the primary mitochondrial disorder. PMID- 19565056 TI - Pancreatic somatostatinoma manifested as severe hypoglycemia. AB - Somatostatinoma is a rare somatostatin-producing endocrine tumor, probably malignant. Due to its nonspecific symptoms such as vague abdominal pain, weight loss, or occult clinical features, misdiagnosis occurs. We report a case of pancreatic somatostatinoma with severe hypoglycemia. The patient had experienced severe hypoglycemic attacks for 11 months periodically. Contrast computed tomography scan revealed an isodensity mass about 2 cm in the head of the pancreas. Ultimately, a local excision was carried out as the tumor was located exactly on the surface of the pancreas. Somatostatinoma was established after immunohistochemical technique. The patient led a normal life without any complaint at 1 year follow-up. PMID- 19565057 TI - Hepatic morphopathologic findings of lead poisoning in a drug addict: a case report. AB - We describe the case of a 40-year old Iranian man who was admitted to our hospital with severe abdominal pain, abnormal liver function tests and normocytic anemia. Suffering from multiple sclerosis, he was a regular user of opium for pain relief. Basophilic stippling of erythrocytes pointed towards the diagnosis of lead intoxication, the most likely source being contaminated Iranian opium. Serum lead and zinc protoporphyrin levels were strongly elevated. To assess the hepatotoxic effects of lead poisoning a liver biopsy was performed. Pathomorphologic findings of hepatotoxicity, rarely reported in humans, included active hepatitis together with extensive microvesicular and macrovesicular steatosis, hemosiderosis and cholestasis, and a lymphocytic cholangitis. Whilst treated with chelating therapy, liver enzymes returned to normal, suggesting reversibility of the histological findings. PMID- 19565058 TI - Heterotopic mesenteric ossification after gastrectomy for watermelon stomach. AB - Heterotopic mesenteric ossification is a rare disorder. Only a few cases have been reported in the literature, associated with previous abdominal surgery or trauma. We report a case of heterotopic mesenteric ossification leading to abdominal sepsis, after abdominal operation for recurrent gastric bleeding, due to gastric antral vascular ectasia (GAVE), otherwise called "watermelon stomach", another rare disorder. PMID- 19565059 TI - Amiodarone induced liver cirrhosis. Report of two cases. AB - Amiodarone is used commonly in patients with cardiac diseases. Common side effects include thyroid dysfunction and hepatic abnormalities. However, recently there has been concern for developing liver cirrhosis secondary to amiodarone therapy. We present two cases of liver cirrhosis in patients taking amiodarone. Their clinical presentation as well as histological features are discussed in detail. PMID- 19565060 TI - Contrast-enhanced ultrasound of the pancreas--a method beyond its potential or a new diagnostic standard? AB - Contrast-enhanced ultrasound is a relatively new technique, currently used for liver tumors diagnosis. Newer contrast agents are composed of stabilized micro bubbles capable of traversing the capillary circulation. Lately, the method has also been used in the assessment of pancreatic disorders. Pulse inversion harmonic imaging allows the assessment of the hypervascularised masses as neuroendocrine tumors, of the hypoperfused masses as adenocarcinomas and of the necrotic areas in acute pancreatitis. Also, this imaging method allows a better assessment of the pancreatic tumor resectability and the identification of septa inside the cystic lesion. Contrast-enhanced ultrasound might represent a valuable additional imaging method to contrast CT for selected cases. PMID- 19565061 TI - Contrast enhanced ultrasound in assessing therapeutic response in ablative treatments of hepatocellular carcinoma. AB - Percutaneous ablative methods guided by imaging techniques are considered nowadays curative treatment for early HCC in patients who are not candidates for liver transplantation and surgical resection. The final goal of all ablative treatments is to achieve complete destruction of neoplastic tissue by disruption of tumor vascularity. The best way to demonstrate the efficacy of any ablative methods noninvasively is to demonstrate that the blood supply has been disrupted both inside and at the periphery of the tumor by means of imaging methods. Contrast-enhanced ultrasound (CEUS) with second generation contrast agents is almost as sensitive as CT (considered to be the gold standard) in depicting the residual tumor after an ablation. Moreover, CEUS can be used before ablation to plan the treatment, during the procedure to guide the needle insertion, or immediately after to determine whether the tumor has been ablated or needs additional treatment which can be performed in the same session. PMID- 19565062 TI - Intraductal papillary mucinous neoplasm with more than one histologic type of epithelium. PMID- 19565063 TI - Plasmodium falciparum and hepatitis B virus co-infection--a rare cause of acute hepatitis. PMID- 19565065 TI - Gallstone ileus as first presentation of a gallbladder carcinoma. PMID- 19565066 TI - Hepatocellular carcinoma in membranous obstruction of the inferior vena cava--a causal or a casual presentation. PMID- 19565067 TI - Acute non-typhoid Salmonella mycotic aneurysm of the thoracic aorta. PMID- 19565068 TI - Pegylated interferon/ribavirin-induced sudden sensorineural hearing loss in a patient with chronic hepatitis C. PMID- 19565069 TI - Metastasis of ulcerative colitis in peristomal skin--an extremely rare case. PMID- 19565070 TI - Magnifying endoscopy with narrow-band imaging or confocal laser endomicroscopy for in vivo rapid diagnostic of Barrett's esophagus. PMID- 19565071 TI - The role of chromogranin A in irritable bowel syndrome. PMID- 19565075 TI - Boronic acids as building blocks for molecular nanostructures and polymeric materials. AB - Boronic acids are versatile building blocks for the construction of complex molecular architectures. Using reversible condensation reactions, it is possible to obtain macrocycles, cages, dendritic structures, and rotaxanes, as well as 1-, 2-, and 3-dimensional polymers in one-pot reactions from simple starting materials. This perspective highlights important recent developments in this area. PMID- 19565076 TI - Amine triphenolate complexes: synthesis, structure and catalytic activity. AB - Triphenolamines are highly modular tetradentate molecules that effectively coordinate to transition metals and main group elements with podand topology. They form chiral complexes with intrinsically well defined coordination geometries controlled by the ligand, in particular by the nature of the substituents in ortho position to the phenol, which are able to influence their reactivity and stability. The metal complexes, especially Ti(iv) and V(v), have been found to be effective catalyst in polymerization reactions and oxygen transfer processes. PMID- 19565077 TI - Oxygen isotopic exchange in an MnIIIMn3IV-oxo cubane. AB - The rate and activation parameters for oxygen exchange of the mu(3)-oxo bridges in a manganese tetranuclear cluster with H(2)(18)O have been measured by ESI-MS; the observed DeltaS(double dagger) of -146 +/- 22 J K(-1.)mol(-1) is consistent with the expected associative mechanism of substitution. PMID- 19565078 TI - The first binucleating amino-thiophenolate macrocycles with pendant hydroxyethyl groups. AB - The synthesis of pendant donor macrocyclic amino-thiophenolate ligands with up to six hydroxyethyl arms has been achieved. The crystal structures of two proligands and a Ni(2)Na(4) complex of one deprotected derivative are reported. PMID- 19565079 TI - Synthesis, characterisation and coordination chemistry of a new multidentate P2N4 ligand system. AB - A new potentially hexadentate P(2)N(4) ligand has been prepared and its coordination chemistry to Pd(II) and Pt(II) investigated. The ligand bonds in a chelating fashion via the diphosphine backbone to PdCl(2) and PtCl(2) while the appended pyridyl groups remain non-bonding. Abstraction of the chloride ions from the platinum complex results in fluxional exchange of the pyridyl groups in solution at room temperature on the NMR timescale. X-ray crystallographic analysis of this platinum complex revealed a tetracoordinate complex with two bound and two free pyridyl arms. PMID- 19565080 TI - A novel non-centrosymmetric metallophosphate-borate compound via ionothermal synthesis. AB - Ionothermal synthesis using a deep eutectic solvent has been used for the first time to prepare a borate-containing material. The metallophosphate-borate compound crystallises in chiral space group I222. Two different alkyl ammonium cations derived from the ionic liquid have distinct template effects on the formation of cobalt phosphate layers and hydrophobic borate channels. PMID- 19565081 TI - Metal complexes with N-(2-pyridylmethyl)iminodiacetate: from discrete polynuclear compounds to 1D coordination polymers. AB - The reactions of M(ClO(4))(2).6H(2)O (M = Mn(2+), Co(2+) and Ni(2+)) with N-(2 pyridylmethyl)-iminodiacetic acid (H(2)pmida) gave three discrete polynuclear complexes {Na[Mn(3)(pmida)(3)(H(2)O)(3)]}ClO(4) (), {Na[Co(3)(pmida)(3)(H(2)O)(3)](H(2)O)}ClO(4) () and [Co(H(2)O)(6)][Co(3)(pmida)(3)(H(2)O)(3)](2)(ClO(4))(2).4H(2)O () and two 1D polymers {[Co(pmida)(H(2)O)].H(2)O}(n) () and {[Ni(pmida)(H(2)O)].H(2)O}(n) (). All complexes were characterized by elemental analysis, infrared (IR) spectroscopy and single-crystal X-ray diffraction analysis. Complexes and show similar structures with an unprecedented trinuclear unit [M(3)(pmida)(3)(H(2)O)(3)] in which the three six-coordinated divalent metal ions are joined by three coplanar syn-anti bridging carboxylate groups of three ligands into a closed planar twelve-membered ring with one Na(+) ion located on the C(3) axis. Complex possesses two kinds of coordination units: mononuclear cation [Co(H(2)O)(6)](2+) and trinuclear unit [Co(3)(pmida)(3)(H(2)O)(3)]. The Co(2+) and Ni(2+) ions in complexes and , respectively, present distorted octahedral geometries and are bridged by anti-anti bridging carboxylate groups of the ligands to form 1D chains. Compounds display a structure variation from discrete polynuclear to 1D coordination polymer along Mn(ii), Co(ii) to Ni(ii), and a dependence of the formation of discrete compound or coordination polymer on the identity of the metal ion. The magnetic investigations of were also carried out. The negative Weiss constants and coupling constants obtained from two kinds of fitting models indicate the presence of dominant antiferromagnetic exchanges mediated by the syn-anti bridging carboxylate groups between the metal centers of three compounds. PMID- 19565082 TI - Syntheses and structures of (N',N'-dimethylhydrazido)silanes. AB - The hydrazidosilanes Ph(2)Si(NHNMe(2))(2) (1), Me(2)Si(NHNMe(2))(2) (2), PhSi(NHNMe(2))(3) (3), MeSi(NHNMe(2))(3) (4), PhClSi(NHNMe(2))(2) (5), MeClSi(NHNMe(2))(2) (6), Me(2)ClSi(NHNMe(2)) (7), MeClHSi(NHNMe(2)) (8), MeHSi(NHNMe(2))(2) (9), Me(2)HSi(NHNMe(2)) (10), Me(2)NN[HSi(NHNMe(2))(2)](2) (11) and Si(NHNMe(2))(4) (12) have been prepared by the reaction of the corresponding chlorosilanes with N,N-dimethylhydrazine. Some of the compounds containing Si-Cl and N-H functions simultaneously (6,7,8 ) are very reactive and tend to polymerise and could only be characterised by spectroscopic methods ((1)H, (13)C, (29)Si NMR, IR, MS). All other compounds could additionally be characterised by elemental analyses. The structures of , , , , and in the solid state were determined by X-ray diffraction. These include the first structural determinations of compounds containing SiH-NH-N (9,11) and SiCl-NH-N (5) units. PMID- 19565083 TI - Syntheses, crystal and electronic structures of compounds AM(PO4)2 (A = Sr, M = Ti, Sn; A = Ba, M = Sn). AB - Three ternary metal diorthophosphates (V), AM(PO(4))(2) (A = Sr, M = Ti, Sn; A = Ba, M = Sn), have been prepared by high-temperature solution growth (HTSG) method and structurally characterized by single-crystal X-ray diffraction analysis for the first time. The structure of BaSn(PO(4))(2) features the yavapaiite-type structure, with space group C2/m and a = 8.214(2), b = 5.2456(13), c = 7.8938(19) A, beta = 94.561(4) degrees , Z = 2, R(1) (all data) = 0.0253. Compounds SrM(PO(4))(2) (M = Ti, Sn) are isostructural and crystallize in a novel distorted yavapaiite structure, with space group C2/c and a = 16.4617(4), b = 5.1720(3), c = 8.1187(2) A, beta = 116.40(2) degrees , Z = 4, R(1) (all data) = 0.016 for SrTi(PO(4))(2); a = 16.674(14), b = 5.223(4), c = 8.099(6) A, beta = 115.821(11) degrees , Z = 4, R(1) (all data) = 0.025 for SrSn(PO(4))(2). Band structure calculations from the density functional theory (DFT) method indicate that all three compounds are approximately direct band-gap insulators. PMID- 19565084 TI - Towards robust alkane oxidation catalysts: electronic variations in non-heme iron(ii) complexes and their effect in catalytic alkane oxidation. AB - A series of non-heme iron(ii) bis(triflate) complexes containing linear and tripodal tetradentate ligands has been prepared. Electron withdrawing and electron donating substituents in the para position of the pyridine ligands as well as the effect of pyrazine versus pyridine and sulfur or oxygen donors instead of nitrogen donors have been investigated. The electronic effects induced by these substituents influence the strength of the ligand field. UV-vis spectroscopy and magnetic susceptibility studies have been used to quantify these effects and VT (1)H and (19)F NMR spectroscopy as well as X-ray diffraction have been used to elucidate structural and geometrical aspects of these complexes. The catalytic properties of the iron(ii) complexes as catalysts for the oxidation of cyclohexane with hydrogen peroxide have been evaluated. In the strongly oxidising environment required to oxidise alkanes, catalyst stability determines the overall catalytic efficiency of a given catalyst, which can be related to the ligand field strength and the basicity of the ligand and its propensity to undergo oxidation. PMID- 19565085 TI - Neutral and cationic main group element cages of germanium(II) with pyrazolyl ligands: solid state structures, DFT calculations and advanced solution NMR investigations. AB - Two equivalents of 3,5-bis(trifluoromethyl)pyrazole or 5-methyl-3 (trifluoromethyl)pyrazole react with [Ge{N(SiMe(3))(2)}(2)] to give dimeric homoleptic germanium(ii) compounds [{Ge(3,5-RR'pz)(2)}(2)] (R = R' = CF(3) (1); R = Me, R' = CF(3) (2)), which provide cationic triply pz-bridged cages of the general formula [Ge(mu(2)-3,5-RR'pz)(3)Ge](+)[OTf](-) (R = R' = CF(3) (3); R = Me, R' = CF(3) (4)) upon treatment with one equivalent of triflic acid (HOTf, HSO(3)CF(3)). The compounds were studied using the pulsed field gradient spin echo (PGSE) NMR technique and (19)F,(1)H-HOESY NMR methods. Single crystal X-ray structure determinations of all compounds are reported. The results obtained are verified by density functional theory calculations. PMID- 19565086 TI - 1,1- and 1,2-isomers of the diborane(4) compound B(2){1,2-(NH)(2)C(6)H(4)}(2) and a TCNQ Co-crystal of the 1,1-isomer. AB - The synthesis and X-ray crystal structures of the diborane(4) isomers 1,1 B(2){1,2-(NH)(2)C(6)H(4)}(2) and 1,2-B(2){1,2-(NH)(2)C(6)H(4)}(2) are described together with the results of quantum chemical calculations which shed light on their relative stabilities and degree of aromaticity. Spectroscopic data are also provided for both isomers of the 4-methyl aryl derivative. The compound 1,1 B(2){1-O-2-(NH)C(6)H(4)}(2) has also been prepared and structurally characterised but no evidence was obtained for the corresponding 1,2-isomer. The compound 1,1 B(2){1,2-(NH)(2)C(6)H(4)}(2) forms a co-crystal with TCNQ, the structure of which is also reported. PMID- 19565087 TI - Structural diversity and properties of Zn(II) and Cd(II) complexes with a flexible dicarboxylate building block 1,3-phenylenediacetate and various heterocyclic co-ligands. AB - A series of Zn(II) and Cd(II) complexes with a flexible dicarboxylate building block and various heterocyclic co-ligands, formulated as {[Zn(2)(pda)(2)(phen)(2)].2H(2)O}(n) (1), {[Zn(pda)(dpe)].H(2)O}(n) (2), [Zn(pda)(bpp)](n) (3), {[Cd(2)(pda)(2)(2,2'-bipy)(2)].2H(2)O}(n) (4), {[Cd(pda)(4,4'-bipy)(H(2)O)].H(2)O}(n) (5) and {[Cd(2)(pda)(2)(bpp)(3)].14H(2)O}(n) (6) (pda = 1,3-phenylenediacetate, phen = 1,10-phenanthroline, dpe = 1,2-di(4-pyridyl)ethylene, bpp = 1,3-bi(4 pyridyl)propane, 2,2'-bipy = 2,2'-bipyridine, and 4,4'-bipy = 4,4'-bipyridine), have been synthesized and structurally characterized. In 1, the (H(2)O)(8) clusters interlink the cyclic coordination dimers to give a 3D network through hydrogen bonding. Both 2 and 3 feature 2D corrugated (4,4) layers, which are of 2 fold interpenetrating for 3. In 4, the dimeric Cd(II) subunits are connected by the 1,3-pda ligands to generate a rampart-shaped 1D chain motif. As for 5, the [Cd(pda)](2) rings are connected by the paired 4,4'-bipy ligands to afford a tube shaped 1D motif. In contrast to 1-5, complex displays a 3D diamond network and, interestingly, the T5(0)A(0)A(2) water tapes are found to locate in the channels of this 3-D array. A structural comparison of these complexes demonstrates that the characteristics of auxiliary ligands (from chelating to bridging) play a key role in governing the coordination motifs as well as the 3-D supramolecular lattices. Solid-state properties such as photoluminescence and thermal stability of 1-6 have also been studied. PMID- 19565088 TI - Interplay of coordinative and supramolecular interactions in formation of a series of metal-organic complexes bearing diverse dimensionalities. AB - A series of new metal-organic coordination complexes, {[Ni(IP)(H(2)O)(4)].(2H(2)O)(SO(4))} (1), [M(IP)(mu(2)-SO(4))(H(2)O)(2)] (M = Zn 2 , and M = Mn 3 ), {[M(2)(IP)(2)(mu(2)-SO(4))(2)(H(2)O)(4)].2H(2)O} (M = Zn 4 , and M = Co 5 ), {[Mn(2)(IP)(mu(2)-SO(4))(H(2)O)(2)].(6.5H(2)O)(sdba)} (6), {[Mn(IP)(2)(H(2)O)(2)][Mn(pydc)(2)(H(2)O)(2)].(4H(2)O)} (7), {[Mn(IP)(fum)(H(2)O)].H(2)O} (8), {[M(3)(IP)(3)(fum)(3)].(8H(2)O)} (M = Cd 9, and M = Zn 10 ), {[Cd(2)(IP)(2)(bptc)(H(2)O)(2)].(7.3H(2)O)} (11), {[Zn(2)(IP)(2)(bptc)(H(2)O)(2)].(3H(2)O)} (12), {Cd(1,4 BDC)(IP)(H(2)O)].(0.5H(2)O)} (13), {[Mn(1,4-BDC)(IP)(H(2)O)].H(2)O} (14), {[Zn(2)(1,4-BDC)(2)(IP)].2.5H(2)O} (15) (IP = 1H-imidazo[4,5-f][1,10] phenanthroline, H(2)sdba = 4,4'-sulfonyldibenzoic acid, H(2)pydc = pyridine-2,6 dicarboxylic acid, H(2)fum = fumaric acid, H(4)bptc = 3,3',4,4' benzophenonetetracarboxylic acid and 1,4-H(2)BDC = 1,4-benzendicarboxylic acid) were prepared via self-assembly of pharmaceutical agent IP with different metal sulfates in the absence/presence of the carboxylate under mild conditions. All these compounds were structurally determined by single-crystal X-ray diffraction. Compounds 2-6 crystallize with N-donor chelating ligands and sulfate anion linkers. Complexes 2-6 possess sulfate anions with mu(2)-bridging modes, the roles of sulfate anions result in 1D chains (2 and 3) and dinuclear units (4, 5and 6). For complex 7 , two different Mn units are chelated by two IP and pydc ligands, respectively. Compound 8 is a 1D chain connected by fum ligands. Polymers 9-12 exhibit 2D network structures composed of tetrametallic and hexametallic clusters. The Cd(ii)/Zn(ii) layers are interdigitated in pairs for complexes 11 and 12 by stacking forces, which exhibit unusual 2D-->3D interdigitated architectures. In 13 and 14, they show 1D undulating and linear chains due to the different coordinate modes of 1,4-BDC. IP firstly acts in a tridentate mode via the two N atoms of pyridyl rings and the third N atom of imidazole ring, and the two carboxylate groups of 1,4-BDC ligand taking three types of coordinative modes connect Zn atoms into a complicated 3D network in 15 . The structural differences among complexes 1-15 indicate the backbone of such dicarboxylate ligands, nature of metal resources and the ratio of metal to ligand play important roles in modulating the formation of the coordination polymers. Thermal stabilities of these crystalline materials, fluorescent properties of these complexes and TD-DFT calculation of spectra of IP were also investigated. PMID- 19565089 TI - Complexation of uranium(VI) with peptidoglycan. AB - We investigated the interaction of UO(2)(2+) with peptidoglycan (PG), the main part of the outer membrane of Gram-positive bacteria, by potentiometric titration and time-resolved laser-induced fluorescence spectroscopy (TRLFS) over a wide pH (2.0 to 9.0) and concentration range (10(-5) to 10(-4) M U(vi), 0.01 to 0.2 g L( 1) PG). With potentiometry two different dissociation constants for the carboxyl sites of glutamic acid and diaminopimelic acid (pK(a) = 4.55 +/- 0.02 and 6.31 +/ 0.01), and one averaged pK(a) for hydroxyl and amino groups (which are not distinguishable) (9.56 +/- 0.03) and the site densities could be identified. With potentiometry three different uranyl PG complexes were ascertained: two 1 : 1 uranyl carboxyl complexes R-COO-UO(2)(+), one with the glutamic acid carboxyl group (log beta(110) = 4.02 +/- 0.03), which has a very small formation ratio, and one with the diaminopimelic acid carboxyl group (log beta(110) = 7.28 +/- 0.03), and a mixed 1 : 1 : 1 complex with additional hydroxyl or amino coordination, R-COO-UO(2)((+))-A(i)-R (A(i) = NH(2) or O(-)) (log beta(1110) = 14.95 +/- 0.02). With TRLFS, also three, but different, species could be identified: a 1 : 1 uranyl carboxyl complex R-COO-UO(2)(+) (log beta(110) = 6.9 +/- 0.2), additionally a 1 : 2 uranyl carboxyl complex (R-COO)(2)-UO(2) (log beta(120) = 12.1 +/- 0.2), both with diaminopimelic acid carboxyl groups, and the mixed species R-COO-UO(2)((+))-A(i)-R (A(i) = NH(2) or O(-)) (log beta(1110) = 14.5 +/- 0.1). The results are in accordance within the errors of determination. PMID- 19565090 TI - Self-assembly of silver(i) and bis-bidentate N,N-donor ligands: from a tetranuclear complex to coordination polymers. AB - The supramolecular self-assembly structures of one tetranuclear silver(i) complex and three silver(i) coordination polymers derived from bis-bidentate imino pyridine ligands and their reduced N-methyl derivatives are discussed. Six new ligands were synthesised, and the X-ray structural analysis of L(1), L(2) and L(5) with silver(i) ions indicated the formation of 1D coordination polymers, namely {[AgL(1)]BF(4)}(n) (1), {[AgL(2)]ClO(4)}(n) (2), and {[AgL(5)]ClO(4).H(2)O}(n) (5), whereas ligand L(4) led to a tetranuclear complex {[AgL(4)]ClO(4)}(4) (4). L(1) is the Schiff-base derived from the condensation of alpha,alpha'-diamino-m-xylene and pyridine-2-carboxaldehyde and L(4) that resulting from the reaction of 2-(4-aminophenyl)ethylamine with 6-methyl-pyridine 2-carboxaldehyde, while L(2) and L(5) are the reduced N-methyl derivatives. The structural determination shows achiral syndiotactic polymers for 1 and chiral isotactic for 2 and 5. The 1D coordination chains of 1 are stabilised by intramolecular pi-pi stacking interactions between the phenyl ring of a spacer with the pyridine of a symmetry related adjacent ligand, while the crystal packing of 5 show inter-chain pi-pi interactions. Compound 4 comprises discrete tetranuclear [AgL(4)](4)(4+) complex cations and the rhomboid disposition of the metal ions leads to a globular arrangement of the silver(i) complex. The results provide an insight about the effect of ligand spacer length and flexibility in stabilising the metallo-supramolecular architecture. PMID- 19565091 TI - Synthesis of a novel bifunctional chelator AmBaSar based on sarcophagine for peptide conjugation and (64)Cu radiolabelling. AB - Copper-64 shows promise as both a suitable PET imaging and therapeutic radionuclide due to its nuclear characteristics. Stable attachment of radioactive (64)Cu(2+) to targeted imaging probes requires the use of a bifunctional chelator. Sarcophagine (Sar) ligands coordinate the metal ion (64)Cu(2+) within the multiple macrocyclic rings comprising the cage structure, yielding extraordinarily stable complexes that are inert to dissociation of the metal ion in vivo. Several (64)Cu labelled RGD derivatives have been applied in imaging of the alpha(nu)beta(3) integrin receptor expression during tumour angiogenesis. In order to design and develop novel molecular imaging probes containing RGD and Sar ligands, we designed a novel versatile Sar cage-like bifunctional chelator named AmBaSar, synthesized using a conventional synthetic strategy. Conjugation with the cyclic peptide RGD, and subsequent labelling with (64)Cu, provided a new PET probe (64)Cu-AmBaSar-RGD for imaging the alpha(nu)beta(3) integrin receptor. PMID- 19565092 TI - Reduction of terphenyl iron(II) or cobalt(II) halides in the presence of trimethylphosphine: an unusual triple dehydrogenation of an alkyl group. AB - The reduction of {ArFeBr}(2) (Ar = terphenyl) with KC(8) in the presence of excess PMe(3) afforded the Fe(i) complex 3,5-Pr(i)(2)-Ar'Fe(PMe(3)) (1) (Ar'-3,5 Pr(i)(2) = C(6)H-2,6-(C(6)H(3)-2,6-Pr(i)(2))-3,5-Pr(i)(2)), which has a structure very different from the previously reported, linear Cr(i) species 3,5-Pr(i)(2) Ar*Cr(PMe(3)) (3,5-Pr(i)(2)-Ar* = C(6)H-2,6-(C(6)H(2)-2,4,6-Pr(i)(3))(2)-3,5 Pr(i)(2)) and features a strong Fe-eta(6)-aryl interaction with the flanking aryl ring of the terphenyl ligand. In sharp contrast, the reduction of {ArCoCl}(2) (Ar = 3,5-Pr(i)(2)-Ar' and Ar') afforded the allyl complexes Co(eta(3)-{1-(H(2)C)(2)C C(6)H(3)-2-(C(6)H(2)-2,4-Pr(i)(2)-5-(C(6)H(3)-2,6-Pr(i)(2)))-3-Pr(i)})(PMe(3))(3) (4) and Co(eta(3)-{1-(H(2)C)(2)C-C(6)H(3)-2-(C(6)H(4)-3-(C(6)H(3)-2,6-Pr(i)(2))) 3-Pr(i)})(PMe(3))(3) (5) formed by an unusual triple dehydrogenation of an isopropyl group. It is proposed that the reduction initially generates an intermediate 3,5-Pr(i)(2)-Ar'Co(PMe(3)), which is similar in structure to , followed by 3,5-Pr(i)(2)-Ar'Co(PMe(3)) decomposition to a cobalt hydride intermediate and dehydrogenation of the isopropyl group via remote C-H activation induced by PMe(3) complexation. Complexes 1, 4, and 5 were characterized by X-ray crystallography. In addition, 1 was studied by NMR and EPR spectroscopy; 4 and 5 were characterized by NMR spectroscopy. PMID- 19565095 TI - Health services research: why it matters to health policy makers and clinicians. PMID- 19565093 TI - Copolymerization of CO2 and cyclohexene oxide using a lysine-based (salen)Cr(III)Cl catalyst. AB - A new, natural lysine-based (salen)Cr(III)Cl ((lys-salen)Cr(III)Cl) complex was prepared and its catalytic activity for the copolymerization of CO(2) and cyclohexene oxide (CHO) was described in the presence of PPNCl (PPN(+) = bis(triphenylphosphoranylidene)ammonium) as cocatalyst. The influence of the reaction time, operating temperature and the molar ratio of the catalyst components on the copolymerization was investigated in detail. The results showed that the (lys-salen)Cr(III)Cl, synthesized from non-ortho-diamine, could effectively catalyze the alternating copolymerization (carbonate linkages = 94.6 99.0%). The selectivity was >95%, and was less sensitive to the temperature and the molar ratio of catalyst components, compared to that of the copolymerization catalyzed by traditional salen-metal complexes. The ESI-MS analyses of oligomer and (lys-salen)Cr(III)Cl indicated that a possible chain-transfer reaction had taken place, which might be induced by the water coordinating to the central metal ion. PMID- 19565096 TI - Venous thromboembolism at the National Healthcare Group, Singapore. AB - INTRODUCTION: Venous thromboembolism (VTE), including its most serious clinical subtype, pulmonary embolism (PE), is a potentially preventable disease. While current assessment tools do not include ethnicity as a risk factor, studies suggest that Asians have lower risk of VTE compared to Caucasians. This study aims to describe 2006 in-hospital and projected population based incidence rates of VTE and PE in Singapore. MATERIALS AND METHODS: Data on 2006 admissions at 3 major NHG hospitals, cases of VTE and their demographics were obtained from the ODS, a large administrative database of the National Healthcare Group (NHG). Demographic characteristics of the 2006 Singapore resident population were obtained from the 2006 Singapore Statistics website. RESULTS: In 2006, there were 860 cases of VTE out of 98,121 admissions in these 3 hospitals. Overall and secondary VTE age adjusted in-hospital burden was 73 and 54 per 10,000 patients, respectively. Caucasians and Eurasians had VTE rates in excess of 100 per 10,000 while Chinese, Malays and Indians each had rates below 100 per 10,000. Assuming that 42.5% of the 2006 Singapore population was served by NHG, the estimated population-based incidence of VTE and PE is 57 and 15 per 100,000, respectively. CONCLUSIONS: As patterns across ethnic groups point to lower VTE rates among Asians compared to Caucasians and Eurasians, analytic studies should be considered to test this hypothesis. There may be a need to develop locally applicable risk assessment tools which can be used to support local guidelines for VTE prophylaxis, thus leading to more acceptable and cost-effective care. PMID- 19565098 TI - Effects of survey mode on results of a patient satisfaction survey at the observation unit of an acute care hospital in Singapore. AB - INTRODUCTION: Over the years, surveys have become powerful tools for assessing a wide range of outcomes among patients. Healthcare managers and professionals now consider patient satisfaction as an outcome by itself. This study aims to determine if results of a patient satisfaction survey are affected by the manner by which the survey instrument is administered. MATERIALS AND METHODS: A patient satisfaction survey was conducted from May 2006 to October 2007 in a tertiary level acute care facility. All patients admitted to the observation unit during the study period were invited to participate. Using a contextualized version of the Consumer Assessment of Healthcare Providers and Systems (CAHPS) Hospital Survey, data was collected through either a phone interview, face to face interview or self-administered questionnaire. Each of these survey modes was administered during 3 different phases within the study period. RESULTS: Eight hundred thirty-two (832) patients were included in the survey. Based on results of univariate analysis, out of the 18 questions, responses to 11 (61.1%) were related to survey mode. Face-to-face interview resulted in the greatest proportion of socially desirable responses (72.7%), while phone interview yielded the highest proportion of socially undesirable responses (63.3%). After controlling for possible confounders, logistic regression results showed that responses to 55.6% of the questions were affected by survey mode. Variations in response between phone interview and self-administered questionnaire accounted for 87.5% of the observed differences. CONCLUSIONS: Researchers must be aware that the choice of survey method has serious implications on results of patient satisfaction surveys. PMID- 19565097 TI - Understanding of diabetes mellitus and health-preventive behaviour among Singaporeans. AB - INTRODUCTION: To study the understanding and perceived vulnerability of diabetes mellitus among Singapore residents, and determine the predictors associated with screening for diabetes mellitus among the people without the condition. MATERIALS AND METHODS: A population-based survey was conducted from December 2004 to October 2005 involving Singapore residents aged 15 to 69 years. Using a standard questionnaire, Health Survey Officers interviewed household members on their understanding and perceived vulnerability of diabetes mellitus and associated cardiovascular risk factors. Data were analysed using SPSS v13. RESULTS: The response rate was 84.5%. Of 2,632 respondents, 291 (11.1%) have diabetes mellitus. Compared to respondents without diabetes, respondents with the disease had better understanding of diabetes and they had favourable health practice of screening for cardiovascular risk factors. Having diabetes mellitus was not associated with a healthier lifestyle. Among non-diabetics, those who had a family history of diabetes had better knowledge and health practices than those who had not. They were significantly more likely to recognise the symptoms and signs (61.5% vs 54.5%) and the causes of diabetes (70% vs 58.2%); and were more likely to have ever tested for diabetes (76.1% vs 60.4%), with P <0.001. Socio demographic characteristics, family history, understanding and perception on the vulnerability of diabetes were identified as predictors associated with health screening for the disease. CONCLUSION: Among all respondents, better understanding was found to be associated with favourable health-preventive behaviours. However, it did not translate into healthier lifestyle. Cultural and socio-demographic profiles must be factored in for any effort on lifestyle modifications. PMID- 19565099 TI - Evaluating user satisfaction with an electronic prescription system in a primary care group. AB - INTRODUCTION: Electronic prescribing has been proposed as an important strategy to reduce medication errors, improve the quality of patient care and create savings in health care costs. Despite these potential advantages, user satisfaction plays a significant role in the success of its implementation. Hence, this study aims to examine users' satisfaction and factors associated with satisfaction regarding an electronic prescription system implemented in the National Healthcare Group Polyclinics in Singapore. MATERIALS AND METHODS: An anonymous survey was administered in October 2007 to all physicians, pharmacists and pharmacy technicians working in the 9 National Healthcare Group Polyclinics. RESULTS: Respondents included 118 doctors and 61 pharmacy staff. The overall level of satisfaction with electronic prescribing was high. Doctors and pharmacists reported a high degree of agreement that electronic prescribing reduces prescribing errors and interventions, and they did not want to go back to the paper-based system. Users were generally satisfied with the functionality of the system but there was some degree of workflow interference particularly for the pharmacy staff. Only 56.9% of the pharmacy respondents expressed satisfaction with the review function of the electronic prescription system and only 51.8% and 60% were satisfied when processing prescriptions that included items to be purchased from an external pharmacy or prescriptions with amendments. The results also revealed that satisfaction with the system was more associated with users' perceptions about the electronic prescription system's impact on productivity than quality of care. CONCLUSION: The survey results indicate that the implementation of the electronic prescription system has gone reasonably well. The survey findings provide opportunities for system and workflow enhancement, which is important as these issues could affect the acceptability of a new technology and the speed of diffusion within an organisation. PMID- 19565100 TI - Impact of pharmacy automation on patient waiting time: an application of computer simulation. AB - INTRODUCTION: This paper aims to illustrate the use of computer simulation in evaluating the impact of a prototype automated dispensing system on waiting time in an outpatient pharmacy and its potential as a routine tool in pharmacy management. MATERIALS AND METHODS: A discrete event simulation model was developed to investigate the impact of a prototype automated dispensing system on operational efficiency and service standards in an outpatient pharmacy. RESULTS: The simulation results suggest that automating the prescription-filing function using a prototype that picks and packs at 20 seconds per item will not assist the pharmacy in achieving the waiting time target of 30 minutes for all patients. Regardless of the state of automation, to meet the waiting time target, 2 additional pharmacists are needed to overcome the process bottleneck at the point of medication dispense. However, if the automated dispensing is the preferred option, the speed of the system needs to be twice as fast as the current configuration to facilitate the reduction of the 95th percentile patient waiting time to below 30 minutes. The faster processing speed will concomitantly allow the pharmacy to reduce the number of pharmacy technicians from 11 to 8. CONCLUSION: Simulation was found to be a useful and low cost method that allows an otherwise expensive and resource intensive evaluation of new work processes and technology to be completed within a short time. PMID- 19565101 TI - Predicting positive blood cultures in patients presenting with pneumonia at an Emergency Department in Singapore. AB - INTRODUCTION: Routine blood cultures have been recommended for all patients in treatment guidelines for community-acquired pneumonia (CAP). This practice has become a major area of resource utilisation, despite the lack of evidence in its clinical utility. Calls for abandoning the practice is balanced by the occasions of uncovering an unexpected pathogen or an unusual antimicrobial resistance pattern. The aim of this study is to identify factors that predict positive blood cultures among patients hospitalised for pneumonia upon presentation at the Emergency Department (ED). MATERIALS AND METHODS: A case control study was carried out on patients treated for pneumonia in the ED who had routine blood cultures performed as part of their management. The pneumonia severity index (PSI) was used to categorize patients into low- and high-risk for 30-day mortality. Logistic regression was carried out to determine factors significantly associated with positive blood cultures, from which a predictive probability equation was used to identify patients whose blood cultures were negative at a pre-determined cut-off, with minimum number of culture positive misclassification. A scoring system was devised, with scores predicting which patients would be likely to have a positive or negative blood culture. RESULTS: A total of 1407 patients with pneumonia were treated at ED from May to December 2006, from whom 1800 blood cultures were performed. Of these, 140 cultures (7.8%) grew organisms, comprising 96 (5.3%) true positive cultures and 44 (2.4%) contaminated cultures. Logistic regression analysis identified ill patients with higher PSI classes, smokers and Malay patients to be more likely to have positive blood cultures. Patients who had prior treatment with antibiotics, chronic obstructive pulmonary disease and cough were less likely to have positive blood cultures. An index to predict a negative blood culture resulted in the accurate classification of all but 4 positive patients while still correctly classifying 27.8% of blood culture negative patients. The area under the ROC curve was 0.71 (95% CI, 0.65-0.76). A simplified scoring system was devised based on the predictive model had a sensitivity of 82% and specificity of 38.2% for a positive blood culture. CONCLUSION: Routine blood cultures yielded negative results in 94% of patients presenting with pneumonia. The development of the clinical scoring system is a first step towards selecting patients for whom blood cultures is performed and improve cost-effectiveness. PMID- 19565102 TI - Profiling acute presenting symptoms of geriatric patients attending an urban hospital emergency department. AB - OBJECTIVE: To study the profile of geriatric patients warded to the emergency department (ED) of an Asian acute care general hospital and determine if they are 'more ill', more likely to have atypical presentations and have a higher utilisation of healthcare resources when compared to a younger group of patients. MATERIALS AND METHODS: This is a retrospective chart review of consecutive patients aged 45 years and above presenting to the study ED over a period of 4 weeks from 4 June 2006 to 1 July 2006. The following data were obtained: (i) demographics, (ii) mode of arrival and triage acuity, (iii) presence of co morbidities, (iv) investigations ordered in the ED, (v) clinical symptoms and diagnoses, (vi) disposition, (vii) length of hospital stay, (viii) injuries and outcomes of elderly fallers. The study population was divided into 2 groups--a study group with patients aged 65 years and above, and a control group with patients aged 45 to 64. RESULTS: There were 2847 patients in the study group and these were compared against 2875 in the control group. Those 65 years and above had greater representation in the ED population compared to the general population. In the study group, the proportion of females, the number arriving by ambulance and the likelihood of having a higher triage acuity increased with age. The elderly had higher rates of co-morbidities. They also had a higher resource utilisation rate. Falls was their commonest presenting complaint. CONCLUSION: It is crucial that EDs recognise the special needs of elderly patients due to the growing ageing population. Healthcare policy makers when allocating resources should take into account the profile of elderly patients presenting to an ED and their resource utilisation. PMID- 19565103 TI - Is EQ-5D a valid quality of life instrument in patients with Parkinson's disease? A study in Singapore. AB - INTRODUCTION: The purpose of the present study was to evaluate the validity of the EQ-5D in patients with Parkinson's disease (PD) in Singapore. MATERIALS AND METHODS: In a cross-sectional survey, patients with PD completed English or Chinese version of the EQ-5D, the 8-item Parkinson's disease questionnaire (PDQ 8), and questions assessing socio-demographic and health characteristics. Clinical data were retrieved from patients' medical records. The validity of the EQ-5D was assessed by testing a-priori hypotheses relating the EQ-5D to the PDQ-8 and clinical data. RESULTS: Two hundred and eight PD patients (English speaking: 135) participated in the study. Spearman correlation coefficients between the EQ 5D and PDQ-8 ranged from 0.25 to 0.75 for English-speaking patients and from 0.16 to 0.67 for Chinese-speaking patients. By and large, the EQ-5D scores were weakly or moderately correlated with Hoehn and Yahr stage (correlation coefficients: 0.05 to 0.43), Schwab and England Activities of Daily Living score (correlation coefficients: 0.10 to 0.60), and duration of PD (correlation coefficients: 0.16 to 0.43). The EQ-5D index scores for patients with dyskinesia or "wearing off" periods were significantly lower than those without these problems. The EQ-5D Visual Analog Scale (EQ-VAS) scores also differed for English-speaking patients with deferring dyskinesia, "wearing off" periods, or health transition status; however, such differences were not observed in patients who completed the survey in Chinese. CONCLUSIONS: The EQ-5D questionnaire appears valid for measuring quality of life in patients with PD in Singapore. However, the validity of EQ-VAS in Chinese-speaking patients with PD should be further assessed. PMID- 19565104 TI - Acceptance of information and communication technologies for healthcare delivery: a SingHealth Polyclinics study. AB - OBJECTIVE: This study aimed to determine the prevalence of short message system (SMS) and internet usage in patients visiting the SingHealth Polyclinics and to measure patients' acceptance of using these technologies in healthcare delivery. MATERIALS AND METHODS: A representative sample of patients visiting the 9 SingHealth Polyclinics were interviewed in-person by trained medical students. Collected information included demographic characteristics, access to and usage of mobile phone/SMS and internet, as well as acceptance and concerns on using these technologies in primary healthcare delivery. RESULTS: Among 705 patients surveyed (mean age: 54.6 years, female: 50.6%, response rate: 92%), 407 (57.7%) were SMS users and 158 (22.4%) were internet users. Two hundred and eighty-four of 412 SMS and/or internet users (40.3% of the entire sample) were comfortable with the use of these technologies in healthcare delivery. Malay or Indian ethnicity, better education, and visiting the clinic for acute symptoms or screening were factors positively associated with willingness to use such technologies. The main concerns associated with the use of SMS and internet in healthcare delivery were preference for in-person consultation with a doctor (23.5%), reduced patient-doctor interaction (23.0%), and increased healthcare cost (20.8%). CONCLUSION: The present prevalence of SMS and internet usage among patients visiting the SingHealth Polyclinics and their concerns towards use of these technologies in healthcare delivery do not support current widespread implementation of services entailing SMS and internet in the study sites. PMID- 19565105 TI - New house-officers' views on unprofessional behaviour. AB - AIM: To determine the views of new house officers (HO) on professionalism and unprofessional behaviour following dismissal in January 2007 of a HO who was caught video-taping nurses in the shower. METHODS: An anonymous self-administered questionnaire was administered during new house officers' orientation. Using a Likert scale (1 = strongly disagree to 5 = strongly agree), HO were asked to rank statements regarding teaching and their understanding of professionalism and professional behaviour, role model-clinicians, their response to 3 real-life examples of unprofessional behaviour, and dismissal and Singapore Medical Council (SMC) registration of the sacked HO. Participation was voluntary. RESULTS: Twenty eight out of twenty-nine (96.6%) international medical graduates (IMG) and 84/95 (88%) house officers who graduated from National University of Singapore (NUS) responded. Their median age was 24 years and 63 of them were male. All IMG compared to 63.1% NUS HO agreed that professionalism was well taught in their medical school (P <0.0001). Majority (82.1%) of IMG compared to 67.9% NUS HO agreed they had adequate role model-clinicians exemplifying professionalism (P <0.0001). Majority (90.8%) of the respondents agreed that the sacked HO's behaviour was not pardonable, a smaller proportion (83.9%) agreed with dismissal but only half (52.7%) agreed that SMC should not register the sacked HO. CONCLUSION: In this study, only two-thirds of NUS HO felt that professionalism was well taught and they had adequate role models. NUS should review this aspect of medical education. Majority of HO agreed with the dismissal but only half felt the misdemeanor was serious enough for SMC not to register the sacked HO. PMID- 19565106 TI - Global trends in cardiology and cardiothoracic surgery--an opportunity or a threat? AB - Coronary heart disease is currently the leading cause of death globally, and is expected to account for 14.2% of all deaths by 2030. The emergence of novel technologies from cardiothoracic surgery and interventional cardiology are welcome developments in the light of an overwhelming chronic disease burden. However, as these complementary yet often competing disciplines rely on expensive technologies, hastily prepared resource plans threaten to consume a substantial proportion of limited healthcare resources. By describing procedural and professional trends as well as current and emerging technologies, this review aims to provide useful knowledge to help managers make informed decisions for the planning of cardiovascular disease management. Since their inception, developments in both specialties have been very rapid. Owing to differences in patient characteristics, interventions and outcomes, results of studies comparing cardiothoracic surgery and interventional cardiology have been conflicting. Outcomes for both specialties continue to improve through the years. Despite the persistent demand for coronary artery bypass surgery (CABG) as a rescue procedure following percutaneous coronary intervention (PCI), there is a widening gap between the numbers of PCI and CABG. Procedural volumes seem to have affected career choices of physicians. Emerging technologies from both disciplines are eagerly awaited by the medical community. For long-term planning of both disciplines, conventional health technology assessment methods are of limited use due to their rapid developments. In the absence of established prediction tools, planners should tap alternative sources of evidence such as changes in disease epidemiology, procedural volumes, horizon scan reports as well as trends in disease outcomes. PMID- 19565107 TI - Development of a diabetes registry to improve quality of care in the National Healthcare Group in Singapore. AB - In Singapore, chronic care is provided by both ambulatory primary care clinics and specialist clinics in hospitals. In 2005, the National Healthcare Group (NHG) embarked to build a diabetes registry to enhance the continuity of care for patients with diabetes and facilitate greater efficiency in outcome measurement. This Chronic Disease Management System (CDMS) links administrative and key clinical data of patients with diabetes mellitus across the healthcare cluster. At the point of patient care, clinicians view a summary of each patient's chronic disease records, consolidated chart with physical parameters, laboratory investigation results and the "patient reminders" listing the clinical decision support prompts when key laboratory and screening tests are due for each patient. The CDMS provides reports of clinical outcomes in a systematic and efficient manner for quality improvement and evidenced-based population management. These include process indicators consisting of the rates of glycated haemoglobin (HbA1c), low-density lipoprotein-cholesterol (LDL-c) and nephropathy tests; and intermediate outcome indicators of the proportion of patients with poor HbA1c (>9%) and optimal LDL-c (<2.6 mmol/L) control. From January 2007 to December 2008, the rates of the 3 process indicators were relatively unchanged and that of HbA1c and LDL-c tests were high. There was gradual improvement in the proportion of patients achieving target level of LDL-c in both primary care clinics and hospitals. Fewer patients at primary care clinics had poorly-controlled HbA1c. As a tool for chronic care delivery, the NHG diabetes registry has made clinical monitoring and outcome management for patients with diabetes mellitus more efficient. PMID- 19565108 TI - Risk adjustment: towards achieving meaningful comparison of health outcomes in the real world. AB - Health outcomes evaluation seeks to compare a new treatment or novel programme with the current standard of care, or to identify variation of outcomes across different healthcare providers. In the real world, it is not always possible to conduct randomised controlled trials to address the issue of comparator groups being different with respect to baseline risk factors for the outcomes. Therefore, risk adjustment is required to address patient factors that may lead to biases in estimates of treatment effects. It is essential when conducting outcomes evaluation of more than trivial significance. Risk adjustment begins by asking 4 questions: what outcome, what time frame, what population, and what purpose. Next, design issues are considered. This involves choosing the data source, planning data collection, defining the sample required, and selecting the variables carefully. Finally, analytical issues are considered. Regression modelling is central to every analytic strategy. Other methods that may augment regression include restriction, stratification, propensity scores, instrumental variables, and difference-in-differences. The construction of risk adjustment models is an iterative process requiring both art and science. Derived models should be validated. Limitations of risk adjustment include reliance on data availability and quality, imperfect method, ineffectiveness when comparators are very different, and sensitivity to different methods used. Thoughtful application of risk adjustment can improve the validity of comparisons between different treatments, programmes and providers. The extent of risk adjustment should be guided by its purpose. Finally, its methodology should be made explicit, so that informed readers can judge the robustness of results obtained. PMID- 19565109 TI - Teaching evidence-based practice: perspectives from the undergraduate and post graduate viewpoint. AB - INTRODUCTION: Evidence-based practice (EBP) involves making clinical decisions informed by the most relevant and valid evidence available. It has been suggested that the outcomes of teaching EBP skills may differ between undergraduates and post-graduates due to different determinants in learning. This paper reviews the current literature and discusses the impact of the teaching environment (undergraduate or post-graduate) for teaching EBP and its impact on EBP competency. METHODS: A search of the literature was performed across the MEDLINE, CINAHL, PsychInfo and ERIC databases. Randomised controlled trials (RCTs) and non randomised trials were eligible for inclusion in the paper. Studies were included for review if they explored the impact of teaching on participants' EBP competency, consisting of critical appraisal skills, knowledge and/or behaviour. RESULTS: Ten articles were eligible for inclusion for this review, of which 7 met all inclusion criteria. EBP competency was shown to increase regardless of whether EBP is delivered to medical students at an undergraduate or post-graduate level. EBP taught to a non-medical undergraduate audience did not modify participants' EBP competency. No study directly compared teaching EBP to an undergraduate and post-graduate audience. CONCLUSIONS: Given the limited amount of studies included in this review, further research incorporating highlevel methodologies is required to establish a clear recommendation on the research question. PMID- 19565110 TI - Practical operations research applications for healthcare managers. AB - Operations research (OR) focuses on the application of analytical methods to facilitate better decision-making. Despite its usefulness and proliferation of papers in the academic literature, there are still major issues around getting OR models widely accepted and used as part of mainstream decision-making by clinicians, health managers and policy-makers. This article aims to raise the awareness of healthcare managers with regard to practical OR applications. PMID- 19565112 TI - Women's health in prison: urgent need for improvement in gender equity and social justice. PMID- 19565113 TI - Fair tests of health-care policies and treatments: a request for help from readers. PMID- 19565114 TI - Rebuilding lives, healing minds. PMID- 19565115 TI - Adolescent pregnancy: a culturally complex issue. PMID- 19565116 TI - Unique, multilingual resource on testing health-care treatments. PMID- 19565118 TI - Swine flu of 1976: lessons from the past. An interview with Dr Harvey V Fineberg. PMID- 19565119 TI - Professional assistance during birth and maternal mortality in two Indonesian districts. AB - OBJECTIVE: To examine determinants of maternal mortality and assess the effect of programmes aimed at increasing the number of births attended by health professionals in two districts in West Java, Indonesia. METHODS: We used informant networks to characterize all maternal deaths, and a capture-recapture method to estimate the total number of maternal deaths. Through a survey of recent births we counted all midwives practising in the two study districts. We used case-control analysis to examine determinants of maternal mortality, and cohort analysis to estimate overall maternal mortality ratios. FINDINGS: The overall maternal mortality ratio was 435 per 100,000 live births (95% confidence interval, CI: 376-498). Only 33% of women gave birth with assistance from a health professional, and among them, mortality was extremely high for those in the lowest wealth quartile range (2303 per 100,000) and remained very high for those in the lower middle and upper middle quartile ranges (1218 and 778 per 100,000, respectively). This is perhaps because the women, especially poor ones, may have sought help only once a serious complication had arisen. CONCLUSION: Achieving equitable coverage of all births by health professionals is still a distant goal in Indonesia, but even among women who receive professional care, maternal mortality ratios remain surprisingly high. This may reflect the limitations of home-based care. Phased introduction of fee exemption and transport incentives to enable all women to access skilled delivery care in health centres and emergency care in hospitals may be a feasible, sustainable way to reduce Indonesia's maternal mortality ratio. PMID- 19565120 TI - Ivermectin versus benzyl benzoate applied once or twice to treat human scabies in Dakar, Senegal: a randomized controlled trial. AB - OBJECTIVE: To compare the effectiveness of oral ivermectin (IV) and two different modalities of topical benzyl benzoate (BB) for treating scabies in a community setting. METHODS: The trial included patients aged 5-65 years with scabies who attended the dermatology department at the Institut d'Hygiene Sociale in Dakar, Senegal. The randomized, open trial considered three treatments: a single application of 12.5% BB over 24 hours (BB1 group), two applications of BB, each over 24 hours (BB2 group), and oral IV, 150-200 microg/kg (IV group). The primary endpoint was the disappearance of skin lesions and itching at day 14. If necessary, treatment was repeated and patients were evaluated until cured. Results were analysed on an intention-to-treat basis. A pre-planned intermediate analysis was carried out after the BB1, BB2 and IV groups had recruited 68, 48 and 65 patients, respectively. FINDINGS: At day 14, 33 patients (68.8%) in the BB2 group were cured versus 37 (54.4%) in the BB1 group and 16 (24.6%) in the IV group (P < 10-6). Bacterial superinfection occurred more often in the IV group than in the BB1 and BB2 groups combined (28% versus 7.8%, respectively; P = 0.006). At day 28, 46 patients (95.8%) in the BB2 group were cured versus 52 (76.5%) in the BB1 group and 28 (43.1%) in the IV group (P < 10-5). These clear findings prompted early study cessation. CONCLUSION: Topical BB was clearly more effective than oral IV for treating scabies in a Senegalese community. PMID- 19565121 TI - Potential lost productivity resulting from the global burden of uncorrected refractive error. AB - OBJECTIVE: To estimate the potential global economic productivity loss associated with the existing burden of visual impairment from uncorrected refractive error (URE). METHODS: Conservative assumptions and national population, epidemiological and economic data were used to estimate the purchasing power parity-adjusted gross domestic product (PPP-adjusted GDP) loss for all individuals with impaired vision and blindness, and for individuals with normal sight who provide them with informal care. FINDINGS: An estimated 158.1 million cases of visual impairment resulted from uncorrected or undercorrected refractive error in 2007; of these, 8.7 million were blind. We estimated the global economic productivity loss in international dollars (I$) associated with this burden at I$ 427.7 billion before, and I$ 268.8 billion after, adjustment for country-specific labour force participation and employment rates. With the same adjustment, but assuming no economic productivity for individuals aged > 50 years, we estimated the potential productivity loss at I$ 121.4 billion. CONCLUSION: Even under the most conservative assumptions, the total estimated productivity loss, in $I, associated with visual impairment from URE is approximately a thousand times greater than the global number of cases. The cost of scaling up existing refractive services to meet this burden is unknown, but if each affected individual were to be provided with appropriate eyeglasses for less than I$ 1000, a net economic gain may be attainable. PMID- 19565122 TI - Setting priorities for global mental health research. AB - OBJECTIVE: To set investment priorities in global mental health research and to propose a more rational use of funds in this under-resourced and under investigated area. METHODS: Members of the Lancet Mental Health Group systematically listed and scored research investment options on four broad classes of disorders: schizophrenia and other major psychotic disorders, major depressive disorder and other common mental disorders, alcohol abuse and other substance abuse disorders, and the broad class of child and adolescent mental disorders. Using the priority-setting approach of the Child Health and Nutrition Research Initiative, the group listed various research questions and evaluated them using the criteria of answerability, effectiveness, deliverability, equity and potential impact on persisting burden of mental health disorders. Scores were then weighted according to the system of values expressed by a larger group of stakeholders. FINDINGS: The research questions that scored highest were related to health policy and systems research, where and how to deliver existing cost effective interventions in a low-resource context, and epidemiological research on the broad categories of child and adolescent mental disorders or those pertaining to alcohol and drug abuse questions. The questions that scored lowest related to the development of new interventions and new drugs or pharmacological agents, vaccines or other technologies. CONCLUSION: In the context of global mental health and with a time frame of the next 10 years, it would be best to fill critical knowledge gaps by investing in research into health policy and systems, epidemiology and improved delivery of cost-effective interventions. PMID- 19565123 TI - Exposure to physical and sexual violence and adverse health behaviours in African children: results from the Global School-based Student Health Survey. AB - OBJECTIVE: To examine associations between exposure to physical violence (PV) or sexual violence (SV) and adverse health behaviours among a sample of children in five African countries. METHODS: In a cross-sectional analysis of data from Namibia, Swaziland, Uganda, Zambia and Zimbabwe - countries that participated in the Global School-based Student Health Survey in 2003 or 2004 - we compared the relative frequency of several adverse health behaviours among children (primarily students 13-15 years of age) who did and who did not report exposure to PV or SV. We estimated odds ratios (ORs) for such behaviours and their 95% confidence intervals (CIs) after adjusting for age and sex. FINDINGS: Exposure to PV during the 12 months preceding the survey was reported by 27-50% (average: 42%) of the children studied in the five countries, and lifetime exposure to SV was reported by 9-33% (average: 23%). Moderate to strong associations were observed between exposure to PV or SV and measures of mental health, suicidal ideation, current cigarette use, current alcohol use, lifetime drug use, multiple sex partners and a history of sexually transmitted infection (P < 0.05 for all associations). For example, the odds of being a current cigarette smoker were higher in children involved in one fight (OR: 2.20; 95% CI: 1.77-2.75), 2-5 fights (OR: 3.43; 95% CI: 2.54-4.63), or 6 fights or more (OR: 5.95; 95% CI: 4.37-8.11) (P for trend < 0.001) during the 12 months preceding the survey than in children unexposed to PV. CONCLUSION: Childhood exposure to PV and SV is common among African children in some countries and is associated with multiple adverse health behaviours. In developing countries, increased awareness of the frequency of exposure to violence among children and its potential health consequences may lead to heightened attention to the need for health promotion and preventive programmes that address the problem. PMID- 19565124 TI - Measles deaths in Nepal: estimating the national case-fatality ratio. AB - OBJECTIVE: To estimate the case-fatality ratio (CFR) for measles in Nepal, determine the role of risk factors, such as political instability, for measles mortality, and compare the use of a nationally representative sample of outbreaks versus routine surveillance or a localized study to establish the national CFR (nCFR). METHODS: This was a retrospective study of measles cases and deaths in Nepal. Through two-stage random sampling, we selected 37 districts with selection probability proportional to the number of districts in each region, and then randomly selected within each district one outbreak among all those that had occurred between 1 March and 1 September 2004. Cases were identified by interviewing a member of each and every household and tracing contacts. Bivariate analyses were performed to assess the risk factors for a high CFR and determine the time from rash onset until death. Each factor's contribution to the CFR was determined through multivariate logistic regression. From the number of measles cases and deaths found in the study we calculated the total number of measles cases and deaths for all of Nepal during the study period and in 2004. FINDINGS: We identified 4657 measles cases and 64 deaths in the study period and area. This yielded a total of about 82 000 cases and 900 deaths for all outbreaks in 2004 and a national CFR of 1.1% (95% confidence interval, CI: 0.5-2.3). CFR ranged from 0.1% in the eastern region to 3.4% in the mid-western region and was highest in politically insecure areas, in the Ganges plains and among cases < 5 years of age. Vitamin A treatment and measles immunization were protective. Most deaths occurred during the first week of illness. CONCLUSION: To our knowledge, this is the first CFR study based on a nationally representative sample of measles outbreaks. Routine surveillance and studies of a single outbreak may not yield an accurate nCFR. Increased fatalities associated with political insecurity are a challenge for health-care service delivery. The short period from disease onset to death and reduced mortality from treatment with vitamin A suggest the need for rapid, field-based treatment early in the outbreak. PMID- 19565125 TI - A comparison of two methods for measuring anti-hypertensive drug use: concordance of use with South African standard treatment guidelines. AB - OBJECTIVE: To assess prescriber adherence to standard anti-hypertensive treatment guidelines in South Africa, determine if supply data are useful indicators of drug use, and assess the cost implications of not complying with the guidelines. METHODS: We undertook two studies: an analysis of records of the anti hypertensive drugs supplied to all 54 public-sector hospitals with a hypertension clinic in KwaZulu-Natal, and a direct-observation survey of anti-hypertensive drug prescriptions presented to pharmacies in a subset of 16 of the 54 hospitals. We calculated the relative use of each anti-hypertensive drug group as a proportion of all anti-hypertensive drugs supplied or prescribed. We ranked drug groups in order of use for comparison with recommended South African standard treatment guidelines, and we compared the proportions derived from supply data with those derived from the prescription survey. FINDINGS: Supply data showed that, in line with treatment guidelines, diuretics and angiotensin-converting enzyme inhibitors were the most frequently supplied medicines (42% and 27%, respectively). However, methyldopa - not included in the treatment guidelines - represented 10% of all anti-hypertensives supplied, but the proportion varied widely between hospitals (0-37%). Reserpine, second choice in the treatment guidelines, was used in high amounts by only two hospitals. Calcium channel blockers and beta blockers represented a small proportion of the anti hypertensive drugs supplied: 6% each. Results from the prescription survey were in concordance with supply data for the most frequently prescribed drugs but gave slightly different estimates of the use of others. CONCLUSION: Supply data, the most available source of information about drug use in developing countries, are (with some provisos) a reliable data source for the evaluation of adherence to treatment guidelines. Our results showed substantial non-adherence to standard treatment guidelines. PMID- 19565127 TI - Apocalypse or redemption: responding to extensively drug-resistant tuberculosis. PMID- 19565128 TI - Biventricular repair in unbalanced atrioventricular septal defect. AB - We present the case of a 2.5 year-old child with unbalanced atrioventricular septal defect due to a small left ventricle (LV) (mitral annulus of 10mm and a 0.4 ratio in relation to the tricuspid annulus, LVDD: 17 mm, LV Vd2: 15 ml/m(2) and LV/RV long-axis ratio of 0.71); he had a favorable outcome after biventricular surgical repair. Normal LV development was observed three months after the operation (mitral annulus of 22 mm, with a 0.84 ratio in relation to the tricuspid annulus, and LVDD of 30 mm). Current parameters for utilization of the hypoplastic ventricle are discussed. PMID- 19565126 TI - Comparative impact assessment of child pneumonia interventions. AB - OBJECTIVE: To compare the cost-effectiveness of interventions to reduce pneumonia mortality through risk reduction, immunization and case management. METHODS: Country-specific pneumonia burden estimates and intervention costs from WHO were used to review estimates of pneumonia risk in children under 5 years of age and the efficacy of interventions (case management, pneumonia-related vaccines, improved nutrition and reduced indoor air pollution from household solid fuels). We calculated health benefits (disability-adjusted life years, DALYs, averted) and intervention costs over a period of 10 years for 40 countries, accounting for 90% of pneumonia child deaths. FINDINGS: Solid fuel use contributes 30% (90% confidence interval: 18-44) to the burden of childhood pneumonia. Efficacious community-based treatment, promotion of exclusive breastfeeding, zinc supplementation and Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae immunization through existing programmes showed cost-effectiveness ratios of 10-60 International dollars (I$) per DALY in low-income countries and less than I$ 120 per DALY in middle-income countries. Low-emission biomass stoves and cleaner fuels may be cost-effective in low-income regions. Facility-based treatment is potentially cost-effective, with ratios of I$ 60-120 per DALY. The cost-effectiveness of community case management depends on home visit cost. CONCLUSION: Vaccines against Hib and S. pneumoniae, efficacious case management, breastfeeding promotion and zinc supplementation are cost-effective in reducing pneumonia mortality. Environmental and nutritional interventions reduce pneumonia and provide other benefits. These strategies combined may reduce total child mortality by 17%. PMID- 19565129 TI - Aortobronchopulmonary fistula in the postoperative period of aortic coarctation. AB - BACKGROUND AND OBJECTIVES: In patients with hemoptysis and a history of aortic surgery, the possibility of aortobronchopulmonary fistula must always be considered. The objective of this study was to report a rare case of hemoptysis due to aortobronchopulmonary fistula in the late postoperative period of aortic surgery. CASE REPORT: Female patient, 34 years, surgical correction of aortic coarctation, presenting massive hemoptysis. The echocardiogram disclosed a pseudoaneurysm. The surgical correction was performed and a Dacron tube graft was implanted in the affected aortic segment successfully. CONCLUSIONS: Aortobronchopulmonary fistulas must be considered in patients with previous aortic surgery, due to the elevated morbimortality if they are not promptly diagnosed and treated. PMID- 19565130 TI - Atypical presentation of rheumatic carditis in pregnancy. AB - We report the case of a 31-year old woman presenting with atypical rheumatic disease in the 28th week of the second pregnancy. Questions related to prevention of the disease and its treatment during the pregnancy-puerperium cycle are discussed. PMID- 19565131 TI - ACC opens arms to Brazilians. PMID- 19565132 TI - Morphological description and clinical implications of myocardial bridges: an anatomical study in Colombians. AB - BACKGROUND: Myocardial bridges represent a hotly debated research topic. Myocardial bridges are considered to be a vascular heart variation due to its intermittent or enduring reducing of the arterial lumen, with a possible ischemic effect. METHODS: 154 hearts were studied, having been extracted as fresh autopsy material. Coronary arteries were injected with synthetic resin and cleaned by extracting the pericardial fat. RESULTS: 92 myocardial bridges were observed in 62 hearts (40.3%). Average myocardial bridge length was 19.9 mm. Myocardial bridges varied per artery from one myocardial bridge in 42 hearts (27.3%), two myocardial bridges in 11 hearts (7.2%), three myocardial bridges in 8 hearts (5.2%) and four myocardial bridges in 1 heart (0.7%). Most myocardial bridges coincided with the anterior interventricular artery in its proximal and intermediate segments in 61 cases (39.6%) and left diagonal artery in 11 cases (7.2%). Arteries emitted just before myocardial bridges (pre-bridge branch) in 50 cases (54.3%) of the sample, with an average caliber of 1.41 mm. A third coronary artery was present in 46 cases (29.8%) of total sample. A higher frequency of myocardial bridges was found in hearts with a third coronary artery. CONCLUSION: It is necessary to consider the clinical importance of the variations considering the arterial distribution and associated clinical implications. PMID- 19565133 TI - Homocysteine and MTHFR and VEGF gene polymorphisms: impact on coronary artery disease. AB - BACKGROUND: Polymorphisms in genes involved in the atherosclerosis development, angiogenesis, and homocysteine (Hcy) metabolism could be risk factors for coronary artery disease (CAD). OBJECTIVE: To evaluate the effect of the VEGF C 2578A and MTHFR C677T polymorphisms on CAD, and the association of these polymorphisms with the severity and extension of atherosclerotic lesions and Hcy concentrations. METHODS: Two hundred and forty-four subjects were evaluated by coronary angiography and included in the study (145 with CAD and 99 controls). The VEGF C-2578A and MTHFR C677T polymorphisms were investigated by the PCR-SSCP and PCR-RFLP techniques, respectively. Plasma Hcy was quantified by liquid chromatography/sequential mass spectrometry (LC-MS/MS). RESULTS: There was no significant difference in allele and genotype distribution between the groups, for both polymorphisms. The univariate analysis showed a higher frequency of the VEGF -2578AA genotype in the group with three-vessel disease (p=0.044). In addition, the VEGF -2578CA genotype was observed more frequently among individuals with <95% stenosis (p=0.010). After adjustment for other risk factors for CAD in a multivariate model, the VEGF C-2578A polymorphism was not found to be an independent correlate of CAD (p=0.688). The MTHFR polymorphism did not show any association with the extension and/or severity of the CAD. The MTHFR C677T polymorphism showed no direct association with hyperhomocysteinemia or increased mean plasma concentrations of Hcy. CONCLUSION: Although there is an apparent association between VEGF C-2578A and the development of coronary atherosclerosis, this association is not independent of conventional cardiovascular risk factors. PMID- 19565134 TI - Use of resting myocardial scintigraphy during chest pain to exclude diagnosis of acute myocardial infarction. AB - BACKGROUND: Images of myocardial perfusion taken during an episode of chest pain have been used for patients in the emergency department. OBJECTIVE: To evaluate the operating characteristics of 99mTc-Tetrofosmin scintigraphy during an episode of chest pain to exclude the diagnosis of acute myocardial infarction. METHODS: One hundred and eight patients admitted with chest pain, or up to four hours after the end of symptoms and nondiagnostic electrocardiogram, underwent resting scintigraphy and measurement of troponin I concentrations. Patients with a history of myocardial infarction (MI) were not excluded (24 patients). Troponin I concentrations were determined at admission and 6 hours later. Nuclear physicians performed a blind analysis of the images, and myocardial infarction was confirmed whenever troponin I level increase was three times that of the control. RESULTS: Resting perfusion image was abnormal in all 6 patients with MI. Only 1 patient had a normal image and increased troponin levels. Fifty-five patients had positive images without MI, and 46 patients had normal images and troponin levels. The prevalence of the disease was 6.5%. The sensitivity and specificity of the resting images during an episode of chest pain to diagnose MI was 85.7% and 45.5%, respectively. The negative predictive value was 97.7%. CONCLUSION: Patients undergoing chest pain protocol with SPECT showed an excellent negative predictive value to exclude diagnosis of myocardial infarction. These results suggest that resting perfusion image is an important tool at the chest pain unit. PMID- 19565135 TI - Effects of an inspiratory muscle rehabilitation program in the postoperative period of cardiac surgery. AB - BACKGROUND: Respiratory muscles are affected after cardiac surgeries. OBJECTIVE: To verify whether the preoperative conditioning of the inspiratory muscles might help to decrease postoperative respiratory dysfunction. METHODS: Thirty volunteers of both genders and with a minimum age of 50 years, while waiting for myocardial revascularization and/or cardiac valve surgery, were randomly assigned to two groups. Fifteen patients were included in a domiciliary program of at least 2 weeks of preoperative training of the inspiratory muscles, using a device with a load corresponding to 40% of the maximum inspiratory pressure. The other 15 patients received general advice and did not train the inspiratory muscle. Spirometry, before and after the training program, as well as the evolution of the arterial blood gases and of the maximum inspiratory and expiratory pressure, before and after the operation were evaluated in both group. The clinical outcomes of the two groups were also compared. RESULTS: We observed that inspiratory muscle training increased the forced vital capacity, the maximum voluntary ventilation and the ratio between the forced expired volume during the first second and the forced vital capacity. The evolution of the arterial blood gases and of the maximum inspiratory and expiratory pressures before and after the operation was similar in both groups, with the outcomes also being similar. CONCLUSION: We concluded that our domiciliary program of inspiratory muscle training was safe and improved the forced vital capacity and the maximum voluntary ventilation, although the clinical benefits of this program were not clearly demonstrable in the present study. PMID- 19565136 TI - Comparison of inflammatory biomarkers between diabetic and non-diabetic patients with unstable angina. AB - BACKGROUND: Studies comparing inflammatory activity between diabetic and non diabetic individuals with acute coronary syndrome are scarce, and none including only patients with unstable angina (UA) has been published to date. OBJECTIVE: We compared serum C-reactive protein (CRP), and interleukin-6(IL-6) between diabetic and non-diabetic patients with unstable angina (UA) to determine if difference in inflammatory activity is responsible for a worse prognosis in diabetic patients. We also evaluated the correlation between inflammatory markers and the metabolic profile in diabetic patients and the correlation between inflammatory response and in-hospital outcomes: death, acute myocardial infarction, congestive heart failure, and length of stay in hospital. METHODS: A prospective cohort study of 90 consecutive patients admitted to a chest pain unit with UA and divided into two groups, diabetic and non-diabetic. Serum CRP, IL-6, metabolic profile and leukocyte count were measured at hospital arrival. RESULTS: Forty-two patients (47%) were diabetic (age 62+/-9) vs. 48 (53%) non-diabetic (age 63+/-12). No differences between median C-reactive protein (1.78 vs. 2.23 mg/l,p=0.74) and interleukin-6 (0 vs. 0 pg/ml,p=0.31) were found between the two groups. There was a positive correlation between CRP and total cholesterol (rs = 0.21,p = 0.05), CRP and LDL-cholesterol (rs=0.22,p=0.04) and between CRP and leukocyte count (rs = 0.32, p = 0.02) in both groups. No associations were found between inflammatory markers and in-hospital outcomes. CONCLUSION: We found no difference in inflammatory activity between diabetic and non-diabetic patients with UA, suggesting that this clinical condition may result in balanced inflammatory activity between the two groups and increase acute-phase proteins independently of metabolic state. PMID- 19565137 TI - Piloerection: a side effect of intravenous administration of dobutamine. AB - BACKGROUND: When a dobutamine stress echocardiogram is performed, piloerection is frequently found in the scalp area. OBJECTIVE: The aim of the present study was to establish the incidence of this phenomenon and its association with other clinical findings. METHODS: A total of 218 consecutive patients that underwent dobutamine stress echocardiography in our department were included in the study. RESULTS: Piloerection was present in 42.2% of the sample. No correlations could be established between piloerection and gender, result of the stress test or other side effects. A statistically significant correlation was established with the age of the patients: piloerection was present in 73% of patients aged 50 years or younger. It appears most often with a dobutamine dose of 10 microg/kg/min. CONCLUSION: Piloerection is a frequent side effect of dobutamine infusion, particularly in patients aged 50 or less; it usually precedes the increase in heart rate caused by dobutamine; therefore, it is an early and clear indication that the intravenous set is working properly. Also, the previous warning of its possible appearance may contribute to the patient's well-being. PMID- 19565138 TI - Left ventricular ejection fraction and volumes as measured by 3d echocardiography and ultrafast computed tomography. AB - BACKGROUND: Real-time three-dimensional echocardiography (RT-3D-Echo) and ultrafast computed tomography (CT) are two novel methods for the analysis of LV ejection fraction and volumes. OBJECTIVE: To compare LVEF and volume measurements as obtained using RT-3D-Echo and ultrafast CT. METHODS: Thirty nine consecutive patients (27 men, mean age of 57+/-12 years) were studied using RT-3D-Echo and 64 slice ultrafast CT. LVEF and LV volumes were analyzed. STATISTICAL ANALYSIS: coefficient of correlation (r: Pearson), Bland-Altman analysis, linear regression analysis, 95% CI, p<0.05. RESULTS: RT-3D-Echo measurements: LVEF ranged from 56.1 to 78.6 (65.5+/-5.58)%; end-diastolic volume ranged from 49.6 to 178.2 (87+/ 27.8) ml; end-systolic volume ranged from 11.4 to 78 (33.1+/-13.6) ml. CT scan measurements: LVEF ranged from 53 to 86% (67.8+/-7.78); end-diastolic volume ranged from 51 to 186 (106.5+/-30.3) ml; end-systolic volume ranged from 7 to 72 (35.5+/-13.4)ml. Correlations between RT-3D-Echo and CT were: LVEF (r: 0.7888, p<0.0001, 95% CI 0.6301 to 0.8843); end-diastolic volume (r: 0.7695, p<0.0001, 95% CI 0.5995 to 0.8730); end-systolic volume (r: 0.8119, p<0.0001, 95% CI 0.6673 to 0.8975). CONCLUSION: Good correlation between LVEF and ventricular volume parameters as measured by RT-3D-Echo and 64-slice ultrafast CT was found in the present case series. PMID- 19565139 TI - Association between leisure-time physical activity and C-reactive protein levels in adults, in the city of Salvador, Brazil. AB - BACKGROUND: Leisure time physical activity (LTPA), defined as any type of bodily movement performed during leisure time, is associated with a reduction in the risk for many cardiovascular injuries. OBJECTIVE: To investigate the existence of an association between leisure time physical activity (LTPA) and C-reactive protein (CRP) levels in adults, in the city of Salvador, State of Bahia, Brazil. METHODS: This was a cross-sectional study, with a sample of 822 men and women, aged > 20 years. Active in leisure time were those with a self-reported practice of physical activities in leisure time; high serum CRP levels were those with values > 3.0 mg/l. Logistic regression analysis was used to compute the odds ratio (OR) with a 95% confidence interval (CI). RESULTS: Using multivariate analysis to adjust for potential confounders, we found an OR of 0.73 (0.68-0.79) among the men which shows the existence of an association between LTPA and high CRP levels only in male individuals. After a stratification by gender, obesity, diabetes and smoking habit, we found an association between LTPA and high CRP in non-obese and non-diabetic male smokers or former smokers; and in obese and non smoking females. CONCLUSION: The results of this study may bring contributions to public health, since they can be used to raise awareness of the importance of LTPA as a prospective strategy for population health improvement. PMID- 19565140 TI - 22q11.2 deletion in patients with conotruncal heart defect and del22q syndrome phenotype. AB - BACKGROUND: The 22q11.2 deletion syndrome is the most frequent human microdeletion syndrome. The phenotype is highly variable, being characterized by conotruncal heart defect, facial dysmorphisms, velopharyngeal insufficiency, learning difficulties and mental retardation. OBJECTIVE: The objective of this study was to investigate the frequency of deletion 22q11.2 in a Brazilian sample of individuals with isolated conotruncal heart defect and 22q11.2 deletion syndrome phenotype. METHODS: Twenty-nine patients were studied by classical cytogenetics, by fluorescence in situ hybridization (FISH), and by molecular techniques. RESULTS: Cytogenetic analysis by G-banding revealed a normal karyotype in all patients except one who presented a 47,XX,+idic(22)(q11.2) karyotype. Using molecular techniques, a deletion was observed in 25% of the patients, all exhibiting a 22q11.2 deletion syndrome phenotype. In none of the cases the deletion was inherited from the parents. The frequency of 22q11.2 deletion was higher in patients with the clinical spectrum of the 22q11.2 deletion syndrome than in patients with isolated conotruncal heart defect. CONCLUSION: Investigating the presence of the deletion and its correlation with the patients' clinical data can help the patients and their families to have a better genetic counseling and more adequate clinical follow-up. PMID- 19565141 TI - Analysis of 6-minute walk test safety in pre-heart transplantation patients. AB - BACKGROUND: The 6-minute walk test (6WT) has been used as a means of assessment of the functional capacity, clinical staging and cardiovascular prognosis. Its safety and metabolic impact have not been frequently described in the literature, especially in patients with severe heart failure with clinical indication for cardiovascular transplantation. OBJECTIVE: To evaluate the occurrence of arrhythmias and cardiovascular changes during 6WT. To correlate 6WT performance with clinical staging and cardiovascular prognosis. METHODS: Twelve patients, 10 of whom males, aged 52 +/- 8 years were evaluated at baseline. 6WT was performed with telemetry electrocardiography, vital signs and lactate monitoring. The patients were followed-up for 12 months. RESULTS: The patients walked 399.4+/ 122.5 (D, m), reaching a perceived exertion (PE) of 14.3+/-1.5 and a 34% baseline heart rate variation. Two patients presented more severe pre-6WT arrhythmia which did not worsen with the exercise, four patients presented a significant increase of blood lactate levels (>5 mmol/dl), and three interrupted the test. The distance walked correlated with the ejection fraction (%) and functional class (NYHA). After 12-month follow-up, three patients died and seven were rehospitalized for cardiac decompensation. The D/PE ratio and 2-minute heart rate recovery (HRR2, bpm) were lower in the death group. CONCLUSION: The clinical and electrocardiographic behaviors suggest that the method is safe, but it may be considered too strenuous for some patients with severe heart failure. Variables related to 6WT performance may be associated with the one-year follow-up mortality. PMID- 19565142 TI - Validation of a new surgical risk score for heart valve surgery: VMCP. AB - BACKGROUND: Some studies have developed scores for the assessment of surgical risk, particularly the EuroSCORE, which, however, is complex and difficult to apply. We suggest a new and simpler score, which is more appropriate for the clinical practice and for the assessment of surgical risk in patients with heart valve diseases. OBJECTIVE: This study was conducted to create and validate a simple and practical score to predict mortality and morbidity related to heart valve surgery. METHODS: Hospital data from 764 patients were collected, and the score was validated using two statistical models: death (= mortality) and length of hospital stay (LHS) > 10 days (= morbidity). The score was composed by four indexes (V [heart valve lesion], M [myocardial function], C [coronary artery disease], and P [pulmonary artery pressure]). A cut-off point was set for the score, and uni and multivariate analyses were performed to confirm whether the score would be able to predict mortality and morbidity. The existence of association with other risk factors was also studied. RESULTS: The score was validated with good internal consistency (0.65), and the best cut-off point for mortality and morbidity was 8. Scores > 8 can predict LHS > 10 days (odds ratio [OR] = 1.7; p = 0.006) and a higher death risk, at least in the univariate analysis (p = 0.049). However, the death risk could not be predicted in the multivariate analysis (p=0.258). CONCLUSION: VMCP scores > 8 can predict LHS > 10 days and may be used as a new tool for the follow-up of patients with heart valve disease undergoing surgery. PMID- 19565143 TI - Postoperative atrial fibrillation after cardiac surgery: who should receive chemoprophylaxis? AB - To evaluate arrhythmogenic risk factors associated with greater incidence of postoperative atrial fibrillation (POAF) in patients undergoing cardiac surgery (CABG and/or valvular surgery) in order to identify those more prone to the development of this arrhythmia for possible chemoprophylaxis. Sixty-six patients who underwent elective cardiac surgery were assessed. The following risk factors for the development of POAF were correlated: advanced age, valvular heart disease (VHD), left atrial (LA) enlargement, left ventricular dysfunction (LVD), electrolyte imbalance (EI), previous CABG, prior use and withdrawal of beta blockers (BB) and/or digitalis 24 hours before surgery). The incidence of AF was high (47%) in our study, most frequently on the first postoperative day. Sixty four percent of the study sample was male, and the mean age was 62 years. Among patients with two or less risk factors for AF, only 24% developed arrhythmia, while the presence of three or more risk factors was associated with increased incidence of postoperative AF (69%), (p = 0.04). Age > 65 years (58% of the patients) was the most prevalent risk factor, followed by LA enlargement in 45% (p = 0.001), and VHD in 38% (p = 0.02). The presence of three or more risk factors increases significantly the incidence of this arrhythmia in the postoperative period after cardiac surgery. Among the primary risk factors are advanced age, left atrial enlargement, and valvular heart disease. PMID- 19565144 TI - Case 1/2009--A 33-year-old man with effort dyspnea and syncope who presented sudden worsening of the dyspnea. PMID- 19565146 TI - [Current subjects on depression]. PMID- 19565147 TI - [Which are the recent clinical findings regarding the association between depression and suicide?]. AB - OBJECTIVE: Suicide is one of the leading causes of mortality worldwide, especially among young subjects. Suicide is considered the outcome of a multidimensional and complex phenomenon, which is a result of the interaction between several factors. The association between psychopathology and suicide has been extensively investigated. Major depression plays an important role among the psychiatric diagnoses associated with suicide. This finding seems to be confirmed by different study designs, and in distinct populations. The present paper aims to briefly review the recent findings regarding the suicide-related clinical features of depression. Moreover, strategies for suicide prevention were also reviewed. REVIEW: Recent references were identified and grouped in order to illustrate the main contributions about depression and suicide. Briefly, the literature review stresses the high prevalence of major depression among subjects presenting suicide behaviors. Psychopathological traits, such as aggression and impulsivity play a relevant role in triggering suicidal behaviors. Strategies for suicide prevention were also reviewed in Brazil and internationally. In general, detection and treatment are effective in reducing suicide rates. CONCLUSION: Studies regarding suicide behaviors have had a pragmatic approach, and generated a large body of evidence about correlates of suicide. However, these studies have not been able to provide a consistent theoretical explanation for this phenomenon. The recent adoption of modern strategies represents a possibility of enhancing the research capability of such studies. In order to be clinically useful, findings should make it possible to deepen the understanding over the experience of a suicidal person, as well as to design specific strategies for prevention and treatment in population subgroups. PMID- 19565148 TI - [Electroconvulsive therapy in major depression: current aspects]. AB - OBJECTIVE: The efficacy of electroconvulsive therapy in treating depressive symptoms has been established by means of innumerable studies developed along the last decades. Electroconvulsive therapy is the most effective biological treatment for depression currently available. The objective of this study was to demonstrate the role of electroconvulsive therapy in the treatment of depression and highlight present aspects related to its practice. METHOD: We reviewed in the literature studies on efficacy, symptom remission, predictive response factors as well as current aspects regarding quality of life, the patients' perception, mechanism of action, technique and cognitive impairment. RESULTS: The main results found in the this revision were: 1) electroconvulsive therapy is more effective than any antidepressant medication; 2) the remission of depression with electroconvulsive therapy varies, in general, from 50 to 80%; 3) The effect of electroconvulsive therapy in brain-derived neurotrophic factor levels is still controversial; 4) electroconvulsive therapy has a positive effect in the improvement of quality of life; 5) patients submitted to electroconvulsive therapy have, in general, a positive perception about the treatment. CONCLUSION: Electroconvulsive therapy remains a highly efficacious treatment in treatment resistant depression. With the improvement of its technique, electroconvulsive therapy has become an even safer and more useful procedure both for the acute phase and for the prevention of new depressive episodes. PMID- 19565149 TI - [Major depression invites major concerns]. AB - OBJECTIVE: To overview limitations to the concept and construct of major depression. METHOD: The objectives in initially conceptualizing major depression are examined against its subsequent utility and relevance to clinicians and researchers. RESULTS: It is argued that, as defined, major depression does not differentiate clinical depression well from expressions of non-clinical depression or sadness, that its criteria set do not generate reliable diagnoses, that a diagnosis of major depression means little in and of itself (as it effectively comprises multiple types of depression) and that it fails to inform us about cause, natural history or differential treatment response. CONCLUSION: Limitations to the concept of major depression would benefit from wider appreciation to advance changes to the clinical diagnosis of depressive sub types. PMID- 19565150 TI - [Transcranial direct current stimulation: a promising alternative for the treatment of major depression?]. AB - OBJECTIVE: In recent years, a number of new somatic (non-pharmacological treatments) have been developed for the treatment of major depression and other neuropsychiatric disorders. Among these, one of the most promising is transcranial direct current stimulation. METHOD: For the present literature review we searched the PubMed between January 1985 and February 2009. To be included, articles should have been published in English and should address general principles of transcranial direct current stimulation and its use in major depression. DISCUSSION: Current protocols for the treatment of major depression with transcranial direct current stimulation usually involve the application of two sponge-electrodes in the scalp. In general, the positive electrode is applied in the region above the left dorsolateral prefrontal cortex (i.e., F3 region of the 10/20 International System for EEG) and the negative electrode is applied in the region above the right supra-orbital area. A direct electrical current of 1-2 mA is then applied between the electrodes for about 20 minutes, with sessions being daily performed for one to two weeks. Initial studies (including a randomized, double-blind, placebo-controlled clinical trial) showed that transcranial direct current stimulation is effective for the treatment of non-complicated major depression and that this technique, when used in depressed patients, is associated with improvement in cognitive performance (including working memory). Finally, transcranial direct current stimulation is safe and well tolerated. CONCLUSION: Recent studies show that transcranial direct current stimulation is an important neuromodulatory method that may be useful for the treatment of depressed patients. However, further studies are needed to better clarify its precise role in the management of depressive disorders. PMID- 19565151 TI - [Review of the guidelines of the Brazilian Medical Association for the treatment of depression (Complete version)]. AB - OBJECTIVE: Depression is a frequent, recurrent and chronic condition with high levels of functional disability. The Brazilian Medical Association Guidelines project proposed guidelines for diagnosis and treatment of the most common medical disorders. The objective of this paper is to present a review of the Guidelines Published in 2003 incorporating new evidence and recommendations. METHOD: This review was based on guidelines developed in other countries and systematic reviews, randomized clinical trials and when absent, observational studies and recommendations from experts. The Brazilian Medical Association proposed this methodology for the whole project. The review was developed from new international guidelines published since 2003. RESULTS: The following aspects are presented: prevalence, demographics, disability, diagnostics and sub diagnosis, efficacy of pharmacological and psychotherapeutic treatment, costs and side-effects of different classes of available drugs in Brazil. Strategies for different phases of treatment are also discussed. CONCLUSION: The Guidelines are an important tool for clinical decisions and a reference for orientation based on the available evidence in the literature. PMID- 19565152 TI - Predicting walking ability following lower limb amputation: a systematic review of the literature. AB - OBJECTIVE: To investigate factors that predict walking with a prosthesis after lower limb amputation. DESIGN: Systematic literature review. METHODS: A computer aided literature search of MEDLINE, EMBASE, CINAHL and the Cochrane Library was performed to identify studies published up to August 2007 that investigated factors that predicted walking ability after lower limb amputation. RESULTS: A total of 57 studies were selected. Predictors of good walking ability following lower limb amputation include cognition, fitness, ability to stand on one leg, independence in activities of daily living and pre-operative mobility. Longer time from surgery to rehabilitation and stump problems are predictors of poor outcome. The impact of the cause of amputation on walking varies between studies. In general, unilateral and distal amputation levels, and younger age were predictive of better walking ability. Sex probably does not have a significant influence on walking ability. CONCLUSION: The heterogeneity of methods and outcome measures used in the identified studies make comparison difficult and, in part, explains conflicting conclusions in relation to predictive factors. Further investigation of predictive factors is needed to estimate walking potential more accurately and guide targeting of modifiable factors to optimize outcome after lower limb amputation. PMID- 19565153 TI - Predictors of pain and function outcome after rehabilitation in patients with patellofemoral pain syndrome. AB - OBJECTIVE: To identify changes in impairments associated with functional and pain outcome in patients with patello-femoral pain syndrome following a standardized physiotherapy treatment. DESIGN: One group pre-post design. SUBJECTS: Seventy four patients (median age 27 years, 52% female) diagnosed with patellofemoral pain syndrome and referred to physiotherapy treatment. METHODS: Baseline measures included self-reported function and pain, which were the dependent variables, and measures of independent variables: strength of quadriceps, hip abduction and hip external rotation; length of hamstrings, quadriceps, plantar flexors, iliotibial band/tensor fascia lata complex, and lateral retinaculum; quality of movement, and fear-avoidance beliefs. Following the baseline measurements, subjects participated in a standardized physiotherapy program. Measurements were repeated at 2-month follow-up. Data were analyzed using 2 forward regression models, the first using function outcome, and the second using pain outcome as the criterion. Age, gender, height and weight were controlled in the regression models. RESULTS: Change in fear-avoidance beliefs about physical activity and change in gastrocnemius length predicted function outcome. Change in fear-avoidance beliefs about physical activity and about work predicted outcome of pain. CONCLUSION: Change in fear-avoidance beliefs about physical activity was the strongest predictor of function and pain outcome. The fact that patients who decreased their fear-avoidance beliefs improved function and decreased pain indicates that perhaps fear-avoidance beliefs should be targeted during the treatment of patients with patellofemoral pain syndrome. PMID- 19565154 TI - Diagnostic accuracy and reliability of muscle strength and endurance measurements in patients with chronic low back pain. AB - OBJECTIVE: Dynamometric trunk muscle strength and endurance tests are performed widely within the rehabilitation management of chronic low back pain. The aim of this study was to examine the accuracy and long-term reliability of these measurements in patients with chronic low back pain. DESIGN: Cross-sectional study. SUBJECTS: Thirty-two patients with chronic low back pain, 19 healthy controls and 15 patients with chronic headache matched for age, sex and body mass index. METHODS: Both patient groups and healthy controls performed isokinetic and isometric trunk extensor and flexor tests on a Biodex 2000 dynamometer. The Biering-Sorensen test served to examine back muscle endurance. Borg-Category Ratio-Scales CR-10 rated participants' body experience immediately before and after the testing. Patients with chronic low back pain repeated measurements after 3 weeks. RESULTS: Among dynamometric tests, isokinetic measurements revealed the best area under the curve (AUC = 0.89) for the discrimination between patients with chronic low back pain and healthy controls. Reliability testing revealed highly significant learning effects for isometric trunk flexion and isokinetic measurements. The Biering-Sorensen test demonstrated excellent accuracy (AUC = 0.93) and no learning effects. Borg-category-ratio-scale ratings were not associated with the observed changes. CONCLUSION: In chronic low back pain dynamometric trunk muscle measures are limited to muscle function assessment purposes. Monitoring treatment outcome in these patients with these measures appears to be problematic because of learning effects. Based on our findings, we recommend the Biering-Sorensen test for management of chronic low back pain rehabilitation. PMID- 19565155 TI - Association between catastrophizing and self-rated pain and disability in patients with chronic low back pain. AB - BACKGROUND: Catastrophizing plays an important role in models of pain chronicity, showing a consistent correlation with both pain intensity and disability. It is conceivable that these associations are mediated or confounded by other psychological attributes. OBJECTIVE: To examine the relative influence of catastrophizing and other psychological variables on pain and disability in patients with chronic low back pain. METHODS: Seventy-eight patients completed the Pain Catastrophizing Scale, Roland Morris Disability Questionnaire, Fear Avoidance Beliefs Questionnaire (work/activity), Modified Somatic Perception Questionnaire, Modified Zung Depression Scale, and Pain Intensity scale. RESULTS: Catastrophizing was significantly correlated with both Pain intensity and Roland and Morris Disability, and with all other psychological variables (all p < 0.001). However, multiple regression analyses showed that Catastrophizing explained no significant variance in Pain intensity beyond that explained by the unique contributions of Modified Somatic Perception and Fear-Avoidance Beliefs (work) and explained no further variance in Disability beyond that explained by the unique contributions of Fear-Avoidance Beliefs (work) and Depression. CONCLUSION: These findings are consistent with previous models proposing that negative psychological attributes are associated with greater perceptions of pain and disability. Nonetheless, our study indicates that measures of catastrophizing show notable measurement overlap in multivariate models. PMID- 19565156 TI - Efficacy of neck stabilization exercises for neck pain: a randomized controlled study. AB - OBJECTIVE: To determine the efficacy of neck stabilization exercises in the management of neck pain. PATIENTS AND METHODS: Sixty patients with neck pain were randomized to 3 groups, as follows: group 1--physical therapy agents including transcutaneous electrical nerve stimulation, continuous ultrasound and infra-red irradiation; group 2--physical therapy agents + isometric and stretching exercises; and group 3--physical therapy agents + neck stabilization exercises. The exercises were performed as a home training programme following a 3-week supervised group exercise. The patients were evaluated with a visual analogue scale, by intake of paracetamol, Neck Disability Index, Beck Depression Scale and range of motion in the 3 planes at baseline and at months 1, 3, 6, 9 and 12. RESULTS: Compared with baseline, all groups showed a significant decrease in visual analogue scale scores during the first 6 months. However, this improvement was maintained only in group 3 at 9 and 12 months, with a significant difference among the groups (p<0.05). During the study, the improvement in disability was marked in group 3 with respect to Neck Disability Index, Beck Depression Scale and range of motion in the frontal plane (p<0.05). CONCLUSION: This study demonstrates the superiority of the neck stabilization exercises, with some advantages in the pain and disability outcomes, compared with isometric and stretching exercises in combination with physical therapy agents for the management of neck pain. PMID- 19565157 TI - Quasi-experimental evaluation of a stress management programme for female county and municipal employees on long-term sick leave due to work-related psychological complaints. AB - OBJECTIVE: To evaluate the effects of a stress management intervention among 24 female patients on sick leave due to work-related psychological complaints. METHODS: The study design was quasi-experimental. All participants received a standard individual treatment for stress at a company healthcare unit. Half of the sample was also enrolled in a group intervention aimed at coping with psychological/somatic symptoms of stress. Data were collected before and after treatment and at 6- and 12-month follow-ups, through questionnaires and blood sampling. Rates of return to work were assessed for up to 5 years. RESULTS: Levels of depression decreased in both conditions and these improvements were maintained at follow-up. The experimental condition was superior in alleviating burnout, as measured with the Karolinska Exhaustion Scale. Paradoxically, these improvements were accompanied by significant increases in levels of glycated haemoglobin in both groups. At 5-year follow-up 40% of participants in both conditions had returned to work. CONCLUSION: The stress management intervention seems to have beneficial effects on self-rated symptoms, but is comparable to treatment as usual with respect to rates of return to work. More studies are needed to investigate the impact of this intervention on physiological parameters. PMID- 19565158 TI - Acoustic analysis of swallowing sounds: a new technique for assessing dysphagia. AB - OBJECTIVE: To perform acoustic analysis of swallowing sounds, using a microphone and a notebook computer system, in healthy subjects and patients with dysphagia affected by neurological diseases, testing the positive/negative predictive value of a pathological pattern of swallowing sounds for penetration/aspiration. DESIGN: Diagnostic test study, prospective, not blinded, with the penetration/aspiration evaluated by fibreoptic endoscopy of swallowing as criterion standard. SUBJECTS: Data from a previously recorded database of normal swallowing sounds for 60 healthy subjects according to gender, age, and bolus consistency was compared with those of 15 patients with dysphagia from a university hospital referral centre who were affected by various neurological diseases. METHODS: Mean duration of the swallowing sounds and post-swallowing apnoea were recorded. Penetration/aspiration was verified by fibreoptic endoscopy of swallowing in all patients with dysphagia. RESULTS: The mean duration of swallowing sounds for a liquid bolus of 10 ml water was significantly different between patients with dysphagia and healthy patients. We also described patterns of swallowing sounds and tested the negative/positive predictive values of post swallowing apnoea for penetration/aspiration verified by fibreoptic endoscopy of swallowing (sensitivity 0.67 (95% confidence interval 0.24-0.94); specificity 1.00 (95% confidence interval 0.56-1.00)). CONCLUSION: The proposed technique for recording and measuring swallowing sounds could be incorporated into the bedside evaluation, but it should not replace the use of more diagnostic and valuable measures. PMID- 19565159 TI - Variations in follow-up services after inpatient stroke rehabilitation: a multicentre study. AB - BACKGROUND: Care after discharge from inpatient stroke rehabilitation units varies across Europe. The aim of this study was to compare service delivery after discharge. METHODS: A total of 532 consecutive patients after stroke were recruited from 4 European rehabilitation centres in Germany, Switzerland, Belgium and the UK. At 2-month intervals, clinical assessments and structured interviews were carried out to document functional status and delivery of services after discharge. Significant factors for receiving follow-up services were analysed using a logistic generalized estimating equation model. RESULTS: After controlling for case-mix, the results showed that Belgian patients were most likely to receive physical therapy but least likely to receive occupational therapy. German patients were least likely to receive nursing care. UK patients were less likely to receive medical care from their general practitioner compared with the other patient groups. CONCLUSION: Clinical characteristics did not explain the variations in service delivery after discharge from in-patient stroke rehabilitation. The decision-making processes involved in the provision of follow up services need to be better documented. To improve our understanding of events post-discharge, the influence of non-clinical factors, such as healthcare regulations, should be explored further. PMID- 19565160 TI - Prognostic value of motor evoked potentials in motor function recovery of upper limb after stroke. AB - OBJECTIVE: To determine the prognostic value of clinical assessment and motor evoked potentials for upper limb strength and functional recovery after acute stroke, and to establish the possible use of motor evoked potentials in rehabilitation. DESIGN: A prospective study. SUBJECTS: Fifty-two patients with hemiparesis were enrolled one month post-stroke; 38 patients concluded the study at 12 months. METHODS: Motor evoked potentials were recorded at baseline and after one month. Upper limb muscular strength (Medical Research Council Scale, MRC) and functional tests (Frenchay Arm Test, Barthel Index) were used as dependent outcome variables 12 months later. Motor evoked potentials were classified as present or absent. Predictive values of motor evoked potentials and MRC were evaluated. RESULTS: At 12 months, patients with baseline recordable motor evoked potentials showed a good functional recovery (positive predictive value 94%). The absence of motor evoked potentials did not exclude muscular strength recovery (negative predictive value 95%). Motor evoked potentials had a higher positive predictive value than MRC only in patients with MRC < 2. CONCLUSION: Motor evoked potentials could be a supportive tool to increase the prognostic accuracy of upper limb motor and functional outcome in hemiparetic patients, especially those with severe initial paresis (MRC < 2) and/or with motor evoked potentials absent in the post-stroke acute phase. PMID- 19565161 TI - Abnormal regional benzodiazepine receptor uptake in the prefrontal cortex in patients with mild traumatic brain injury. AB - OBJECTIVE: The diagnosis and management of mild traumatic brain injury continues to be a subject of debate, with varying opinions regarding the extent to which organically based impairments vs the impact of other stressors cause ongoing disability. The aim of this study was to elucidate the possible abnormalities in benzodiazepine receptor uptake in the brains of patients with mild traumatic brain injury. Nine unmedicated patients with mild traumatic brain injury were investigated using 123I-iomazenil single photon emission computerized tomography (SPECT). DESIGN: A descriptive study comparing patients after mild traumatic brain injury with matched control subjects. SUBJECTS: Nine patients with mild traumatic brain injury and 5 controls. METHODS: The SPECT scan was taken 180 min after injection of tracer. RESULTS AND CONCLUSION: All 9 patients had a significant increase (> 2 standard deviations higher than the mean of controls) in benzodiazepine receptor uptake in the prefrontal cortex and significantly higher frontal association cortex-to-average global brain activity ratios than in controls. This SPECT study demonstrated focally altered benzodiazepine receptor uptake in the prefrontal cortices in patients with mild traumatic brain injury. PMID- 19565162 TI - Functional electrical stimulation-assisted cycling of patients with multiple sclerosis: biomechanical and functional outcome--a pilot study. AB - OBJECTIVE: To determine whether functional electrical stimulation-supported ergometric training of patients with multiple sclerosis has a prosthetic or therapeutic effect on biomechanical (power, smoothness of cycling) and functional outcomes (walking capability, strength of muscle, spasticity). DESIGN: Twelve subjects with multiple sclerosis participated in an electrical stimulation supported ergometric training (3 sessions/week for 2 weeks). Measurements were made in a cross-over design to study prosthetic (with and without stimulation) and therapeutic effects (before and after training). METHODS: Power and smoothness were calculated by cadence and torque recordings of cycling and spasticity; strength and walking capability were measured by the Modified Ashworth Scale, Manual Muscle Test, and 10-Metre Walk Test. RESULTS: The power and smoothness of pedalling significantly improved prosthetically with electrical stimulation (p=0.02), but did not show significant improvement over the 2 weeks of training. Significant short-term reductions in spasticity (before vs after training session; p<0.05) were found. Isometric strength did not increase significantly during the 2-week training period and there was no improvement in walking ability. CONCLUSION: Patients with multiple sclerosis are able to improve their cycling power and smoothness by pedalling with stimulation. We suggest that severely affected patients benefit more from functional electric stimulation cycling therapy than do slightly affected patients. PMID- 19565163 TI - Skin disorders in patients with hemiplegia and papaplegia. AB - OBJECTIVE: The aim of this study was to determine the incidence of skin disorders in patients with hemiplegia and paraplegia. Several skin disorders have been reported previously in these patients. METHODS: Seventy inpatients with hemiplegia, 30 with paraplegia, and 90 individuals as a control group were included in the study. RESULTS: The most common skin disorder in the patient group was tinea pedis, which was observed in 18 of the 100 patients. The other common dermatological disorders in the patient group were onychomycosis of the toenails (n = 14), xerosis of the extremities (n = 13) and reduction in hair on the lower extremities (n = 12). The incidence of tinea pedis (p = 0.004), onychomycosis of the toenails (p = 0.010), xerosis of the extremities (p = 0.017) and reduction in hair on the lower extremities (p = 0.027) in the patient group were significantly more common than in the control group. There was no significant correlation between tinea pedis, onychomycosis of the toenails, xerosis of the extremities and reduction in hair on the lower extremities and the duration of hemiplegia and paraplegia. CONCLUSION: Dermatological disorders are observed more commonly in hemiplegic and paraplegic patients than in controls, therefore routine dermatological examination should be performed in these patients. PMID- 19565164 TI - Inpatient rehabilitation services for patients after stroke in Thailand: a multi centre study. AB - OBJECTIVE: To set a baseline for measuring the development of medical rehabilitation services and outcomes for patients after stroke and promoting rehabilitation medicine in Thailand. DESIGN: Multi-centre, prospective study. SUBJECTS: Patients after stroke in Thai Stroke Rehabilitation Registry (TSRR), aged > or = 18 years, with stable medical signs, able to follow a 1-step command and sit for at least 30 min. METHODS: Rehabilitation services were recorded daily as units of treatment (1 unit = 20 min). Length of stay and treatment costs were calculated. RESULTS: Of 327 patients enrolled in the study, 285 (87.2%) completed their treatment programme. Mean length of stay was 29.4 (standard deviation 17.9) days. The average number of total treatment units for stroke rehabilitation was 319.5 (range 27-1674 units), with 205 units of nursing, 40 units of physical therapy and 34 units of occupational therapy as the top 3 services provided. The mean total cost for all treatments during rehabilitation was 28,399 (standard deviation 22,511) baht (approximately USD 789). The ratio of costs not related to rehabilitation to those related to rehabilitation was approximately 2:1. CONCLUSION: This study reports the baseline for measuring the development of rehabilitation services for patients after stroke with a mean length of stay of one month, and for estimating the reasonable costs. PMID- 19565165 TI - Food for thought... on evidence-based toxicology. PMID- 19565166 TI - The suitability of BV2 cells as alternative model system for primary microglia cultures or for animal experiments examining brain inflammation. AB - The role of microglia in neurodegeneration, toxicology and immunity is an expanding area of biomedical research requiring large numbers of animals. Use of a microglia-like cell line would accelerate many research programmes and reduce the necessity of continuous cell preparations and animal experimentation, provided that the cell line reproduces the in vivo situation or primary microglia (PM) with high fidelity. The immortalised murine microglial cell line BV-2 has been used frequently as a substitute for PM, but recently doubts were raised as to their suitability. Here, we re-evaluated strengths and potential short-comings of BV-2 cells. Their response to lipopolysaccharide was compared with the response of microglia in vitro and in vivo. Transcriptome (480 genes) and proteome analyses after stimulation with lipopolysaccharide indicated a reaction pattern of BV-2 with many similarities to that of PM, although the average upregulation of genes was less pronounced. The cells showed a normal regulation of NO production and a functional response to IFN-gamma, important parameters for appropriate interaction with T cells and neurons. BV-2 were also able to stimulate other glial cells. They triggered the translocation of NF-kappaB, and a subsequent production of IL-6 in astrocytes. Thus, BV-2 cells appear to be a valid substitute for PM in many experimental settings, incuding complex cell-cell interaction studies. PMID- 19565167 TI - Quantitation of angiogenesis and antiangiogenesis in vivo, ex vivo and in vitro - an overview. AB - Angiogenesis is defined as the sprouting of new capillaries from pre-existing ones. Angiogenesis is a prerequisite for growth and differentiation of organs and tissues and is involved in many pathological processes, for example growth and metastasis of tumours. Numerous in vivo and in vitro models of angiogenesis have been developed in order to identify and analyse pro- and antiangiogenic factors. This review aims at outlining the topic of quantitation of angiogenesis and antiangiogenesis in these models. PMID- 19565168 TI - Animal and non-animal experiments in nanotechnology - the results of a critical literature survey. AB - A literature survey funded by the Foundation Animalfree Research was performed to obtain an overview on animal experiments in nanotechnology. Scientific articles from Germany, France, the United Kingdom, Italy, the Netherlands and Switzerland published between 2004 and 2007 were collected. A total of 164 articles was retrieved covering in vivo nanotechnological research. The majority of animal experiments were conducted in "nanomedicine", i.e. nanotechnology in the health care area, to study targeted drug, vaccine or gene delivery. Further areas of research relate to nanotechnology-based imaging technologies, the toxicity of nanomaterials, tissue engineering for regenerative treatments, and magnetic tumour thermotherapy. Many experiments were classified as moderately and even severely distressful to the animals. Due to the significance of the scientific topics pursued, the possible scientific benefit of the research depicted in the articles is also assigned to be moderate to high. Nevertheless, it has to be asked whether such animal experiments are truly the only means to answer the scientific questions addressed in nanotechnology. An overview on non-animal test methods used in nanotechnological research revealed a broad spectrum of methodologies applied in a broad spectrum of scientific areas, including those for which animal experiments are being performed. Explicit incentives to avoid animal experiments in nanotechnology currently can only be found in the area of nanotoxicology, but not in the area of nanomedicine. From the point of view of animal welfare, not least because of the new technologies that arise due to nanotechnology, it is time for a paradigm change both in fundamental and applied biomedical research to found research strategies on non-animal test methods. PMID- 19565169 TI - Miticide residues in Virginia honeys. AB - Fifty honey samples from Virginia USA were analyzed for the presence of fluvalinate and coumaphos residues. Samples were collected from hives and from bottled honey provided by beekeepers. No coumaphos or fluvalinate residues above the limit of quantification (0.05 mg/kg) were detected in any of the samples, although trace levels (<0.05 mg/kg) of coumaphos were detected in three samples from hives and trace levels of fluvalinate were found in one hive sample. No residues were detected in any of the bottled honey samples and none of the samples exceeded the US EPA tolerance levels for either miticide. PMID- 19565170 TI - Determinations of MC-LR and [Dha(7)] MC-LR concentrations and physicochemical properties by liquid chromatography-tandem mass spectrometry. AB - A liquid chromatography electrospray mass spectrometry (LC/ESI/MS) method working in multiple reactions monitoring mode for the determination of trace amounts of microcystin variants (MC-LR and [Dha(7)] MC-LR) in waters was developed. The limit of quantification was 0.05 microg/L and the limit of detection was 0.015 microg/L for MC-LR and [Dha(7)] MC-LR, respectively. Recoveries for MCs were in the range of 68%-81%. MC-LR and [Dha(7)] MC-LR were chemically stable with similar physiochemical behavior. PMID- 19565171 TI - Concentration, distribution and sources of polyaromatic hydrocarbons in soils from the Karst tiankengs, South China. AB - Polycyclic aromatic hydrocarbons (PAHs) were investigated in the surface soils from Dashiwei and Datuo Karst tiankengs, Guangxi Province, South China. Concentrations of total PAHs ranged from 16.93 ng g(-1) to 190 ng g(-1) with a mean of 58.29 ng g(-1). In Dashiwei more PAHs were concentrated than in Datuo (mean 74.42 ng g(-1) > 42.15 ng g(-1)). Dashiwei and Datuo accounted for the majority of all PAHs concentrated in the lower location through increased rates (58% and 37%) from the upper to lower geographical levels, respectively. Dashiwei presented the stronger trapping ability for heavier molecular weights on the tiankeng floor with a concentration increase rate of 167% when compared to that of the lighter molecular weights (which had a concentration increase of rate 6%), which was in contrast to the Datuo with a concentration increase rate of 37% for lighter molecular weights and 31% for heavier molecular weights. In Dashiwei, it was found that the ratio of heavier PAH compound molecular weight resulted in its higher precipitation recording on the tiankeng floor. The differentia between two tiankengs, therefore, are related to their environmental characteristics and the PAHs physiochemical properties. The analytical result from the PAHs indicators suggest that a petrogenic source was dominant in the area studied. Therefore, the karst tiankeng has the potential to act as a geophysical well for PAHs deposition, especially for the heavier molecular weighted PAH. PMID- 19565172 TI - Effects of organic-rich sediment and below-ground sulfide exposure on submerged macrophyte, Hydrilla verticillata. AB - The effects of organic-rich sediment and sulfide exposure on Hydrilla verticillata were investigated. The organic richness of sediment was simulated by adding sucrose into sediments, and sulfide exposure was conducted by adding sodium sulfide to plant roots. The length, biomass and density of shoot reduced in the sucrose-amended sediments, and the largest reduction occurred in the highest 1.0% addition treatment by 84.2%, 56.7% and 92.4%, respectively. However, the 0.1% addition treatment stimulated the growth of root. The effects of below ground sulfide exposure on the physiological activities of H. verticillata were determined by adding sulfide to the below-ground tissue. Significantly inhibitory effects of sulfide were observed on plant photosynthesis, root carbohydrate and nitrogen synthetic reserves. The net photosynthetic rates, soluble carbohydrate and soluble protein contents in root were reduced by 104%, 71.8% and 49.8%, respectively, in the 0.6 mM sulfide treatment. PMID- 19565173 TI - Environmental and climatic factors associated with epizootic ulcerative syndrome (EUS) in fish from the Zambezi floodplains, Zambia. AB - This study was conducted to determine environmental and climatic factors associated with Epizootic Ulcerative Syndrome (EUS) in fish in the Zambezi floodplains. EUS is a fish disease that causes economic loses to the fishing industry. Streambed colour in affected water was rusty-, reddish- or yellowish- brown and pH 4.5-6.0 while pH of non affected water was 7.2. The rusty-brown precipitate on fish gills was positive for Prussian blue iron stain. Therefore, predisposing factors for EUS in the Zambezi floodplains were the acidification of ground water during drought years and eventual contamination of surface water during the floods of 2006/2007. PMID- 19565174 TI - The relationship between low bone mass and metabolic syndrome in Korean women. AB - SUMMARY: We examined the relationship between low bond mass and metabolic syndrome in 2,475 Korean women. After adjustment for all covariates, mean vertebral BMD was significantly lower in women with metabolic syndrome. Moreover, age and weight adjusted vertebral BMD was significantly decreased with additional components of the metabolic syndrome. INTRODUCTION: Obesity-induced chronic inflammation is a key component in the pathogenesis of insulin resistance and metabolic syndrome. It has been suggested that proinflammatory cytokines and low grade systemic inflammation activate bone resorption and may lead to reduced bone mineral density (BMD). The objective of this study was to determine the relationship between low bone mass and metabolic syndrome in Korean women. METHODS: This is a cross-sectional study of 2,548 women aged 18 years and over who had visited the Health Promotion Center. Physical examination and laboratory tests were performed. Vertebral BMD was measured using dual-energy X-ray absorptiometry. Metabolic syndrome was defined by National Cholesterol Education Program-Adult Treatment Panel III criteria. RESULTS: Among 2,475 women, 511 (21.0%) women had metabolic syndrome. Women with abdominal obesity or hypertriglyceridemia had significantly lower vertebral BMD than women without respective components after adjustment for age, weight, and height. After adjustment for all covariates, mean vertebral BMD was significantly lower in women with metabolic syndrome (p = 0.031). Moreover, age- and weight-adjusted vertebral BMD were significantly decreased with additional components of the metabolic syndrome (p = 0.004). CONCLUSIONS: These findings suggest that metabolic syndrome might be another risk factor for osteoporosis and related fractures. PMID- 19565175 TI - The cost-effectiveness of risedronate in the UK for the management of osteoporosis using the FRAX. AB - SUMMARY: The study estimated the cost-effectiveness of risedronate compared to no treatment in UK women using the FRAX algorithm for fracture risk assessment. A Markov cohort model was used to estimate the cost-effectiveness. Risedronate was found cost-effective from the age of 65 years, assuming a willingness to pay for a QALY of 30,000 pounds. INTRODUCTION: The aim of this study was to assess the cost-effectiveness of risedronate for the prevention and treatment in a UK setting using the FRAX algorithm for fracture risk assessment. A further aim was to establish intervention thresholds with risedronate treatment. METHODS: The cost-effectiveness of risedronate was compared to no treatment in post-menopausal women with clinical risk factors for fracture using a Markov cohort model populated with data relevant for the UK. The model incorporated the features of FRAX (the WHO risk assessment tool). The analysis had a health care perspective and quality adjusted life years was used as the main outcome measure. RESULTS: Treatment was cost-effective from the age of 65 years, assuming a willingness to pay for a QALY of 30,000 pounds. Treatment was also cost-effective at all ages in women who had previously sustained a fragility fracture or in women with a parental history of hip fracture with a bone mineral density set at the threshold of osteoporosis. At the 30,000 pounds threshold value for a QALY, risedronate was on average found to cost-effective below the 10-year probability of a major osteoporotic fractures of 13.0%. CONCLUSIONS: Risedronate is a cost-effective agent for the treatment of established osteoporosis (osteoporosis and a prior fragility fracture) in women from the age of 50 years and older and above 65 years in women with osteoporosis alone. The results support the treatment recommendations in recent UK guidelines for osteoporosis. PMID- 19565176 TI - Population screening for osteoporosis risk: a randomised control trial of medication use and fracture risk. AB - Randomised control trial of osteoporosis screening in 4,800 women aged 45-54 years was carried out. Screened group observed an increase of 7.9% in hormone replacement therapy (HRT) use (p < 0.001), 15% in other osteoporosis treatments (p < 0.001) and a 25.9% reduction in fracture risk compared with control. Screening for osteoporosis significantly increases treatment use and reduces fracture incidence. INTRODUCTION: Population screening programmes can identify menopausal women with low bone mineral density (BMD) and elevated risk of future fracture but require to be proven effective by a randomised control trial. METHODS: A total of 4,800 women, 45-54 years, were randomised in equal numbers to screening or no screening (control) groups. Following screening, those in the lowest quartile of BMD were advised to consider HRT. Nine years later, the effect of screening on the uptake of treatment and the incidence of fractures were assessed by postal questionnaire. Categorical differences were assessed using chi(2) test. Cox regression was used to assess hazard ratio (HR). RESULTS: Of the screened and the control groups, 52.4% vs 44.5%, respectively, reported taking HRT (p < 0.001). In addition, 36.6% of the screened vs 21.6% of the control groups reported the use of vitamin D, calcium, alendronate, etidronate or raloxifene (p < 0.001). In a per protocol analysis of verified incident fractures, a 25.9% reduction in risk of fractures (of any site) in the screened group was observed (HR = 0.741, 95% CI = 0.551-0.998 adjusted age, weight and height). CONCLUSIONS: Screening for osteoporosis as assessed by low bone density significantly increases the use of HRT and other treatments for osteoporosis and reduces fracture incidence. PMID- 19565177 TI - No increase in adverse pregnancy outcomes for women receiving antiepileptic drugs. AB - The objectives of this study were (1) to investigate the incidence of adverse pregnancy outcomes among the epileptic and general populations, including small for gestational age (SGA), low birth weight (LBW) and preterm delivery, using two large-scale nationwide population-based databases, and (2) to compare the risk of these adverse pregnancy outcomes between epileptic women who did and who did not receive antiepileptic drug (AED) treatment during pregnancy. This study used two national datasets: the National Health Insurance Research Dataset and birth certificate registry. We identified a total of 1,182 women who gave birth from 2001 to 2003 in Taiwan who had been diagnosed with epilepsy within the 2 years preceding the index delivery, together with 5,910 matched women as a comparison cohort. Multivariate logistic regression analyses were performed for estimation of risk. We found that approximately 14% of women with epilepsy received AED treatment during gestation. The adjusted odds of LBW, preterm births and SGA for epileptic women not receiving AED treatment during pregnancy were 1.31 (95% CI, 1.02-1.68), 1.35 (95% CI, 1.07-1.71) and 1.23 (95% CI, 1.03-1.46) times that of women without epilepsy, respectively. In contrast, no significant difference in the risk of LBW infants, preterm births and SGA babies was observed between epileptic mothers receiving AED treatment during pregnancy and women without epilepsy. Our study documents an increased risk of adverse pregnancy outcomes for epileptic women who do not receive AED treatment during pregnancy, but none for epileptic women who do receive treatment. PMID- 19565178 TI - Jules Froment (1878-1946). PMID- 19565179 TI - Plasma tissue factor pathway inhibitor levels in patients with acute pancreatitis. AB - BACKGROUND: In acute pancreatitis (AP), disorders of the coagulation-fibrinolysis system are closely related to the severity of the AP and to organ dysfunctions. We previously reported that plasma tissue factor (TF) levels were elevated in patients with AP, particularly in cases of alcoholic AP with pancreatic necrosis. Tissue factor pathway inhibitor (TFPI) is a key regulator of the extrinsic coagulation pathway, but plasma TFPI levels in AP have not yet been determined. METHODS: Plasma TFPI concentrations were measured by enzyme-linked immunosorbent assay in 44 patients with AP on admission. The relationships between AP severity, pancreatic necrosis, organ dysfunction, infection, and prognosis were analyzed. RESULTS: Plasma TFPI levels were increased in AP patients compared with healthy volunteers. Plasma TFPI levels in severe AP were greater than those in mild AP. Plasma TFPI levels significantly correlated with Ranson score, APACHE II score, and Japanese severity score. Plasma TFPI levels in patients with pancreatic necrosis were greater than those in patients without pancreatic necrosis. Plasma TFPI levels in patients with organ dysfunction were greater than those in patients without organ dysfunction. In patients with pancreatic necrosis, the TF/TFPI ratios in non-survivors were lower than those in survivors. Moreover, the mortality rates in patients with TF/TFPI ratios > or = 2.0 were lower than those in patients with TF/TFPI ratios < 2.0. CONCLUSIONS: Plasma TFPI levels were significantly increased in patients with AP, and the elevation was markedly related to the severity, pancreatic necrosis and organ dysfunctions. The imbalance of TF and TFPI may influence the disease state and thereby the prognosis in AP. PMID- 19565180 TI - A chronic myeloid leukemia patient with atypical karyotype and BCR-ABL e13a3 transcript caused by complex chromosome rearrangement. AB - Philadelphia (Ph) chromosome as a result of t (9; 22) (q34; q11) is observed in more than 90% of chromic myeloid leukemia (CML) patients. Cases in which the typical Ph chromosome is not visible at the karyotype level comprise 5-10% of CML patients. CML cases with fusion transcripts such as e13a3 in which ABL exon 3 rather than exon 2 has fused to BCR are very rare. Such reported cases with the e13a3 transcript show the Ph chromosome on karyotype analysis. We reported an atypical karyotype CML patient with the e13a3 BCR-ABL transcript caused by complex translocation. Fluorescence in situ hybridization (FISH) analysis of the metaphase led to a precise cytogenetical characterization. The patient showed favorable response to imatinib, and achieved major molecular responses. PMID- 19565181 TI - Increased serum brain-derived neurotrophic factor protein upon hypoxia in healthy young men. AB - Exposing animals to brief hypoxic periods leads to neuroprotective ischemic tolerance termed preconditioning. This phenomenon is well documented in the brain, but the underlying mechanisms require further elucidation. As nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are important mediators of maintaining homeostatic conditions in the adult nervous system in terms of physiological and pathophysiological processes, we hypothesized that hypoxic preconditioning might modulate serum neurotrophin concentrations. Hypoxia was induced for 30 min in 14 healthy young men resulting in a constant blood oxygen saturation of 75%. Hyperinsulinemic euglycemic clamps were performed and serum concentrations of BDNF and NGF were measured at baseline, directly after the intervention, and at the end of the session. Overall, serum BDNF concentrations decreased over time by maximally 35% (P = 0.001) while in contrast NGF concentrations remained unchanged. Acute hypoxia alleviated the decrease of BDNF resulting in higher BDNF concentrations as compared to normoxic control (P < 0.01). Our findings show that acute hypoxia results in significantly higher serum BDNF concentrations pointing to a potential role of BDNF in the underlying mechanism of hypoxic preconditioning. Based on its neuroprotective properties, BDNF may be of high clinical relevance for therapeutic options in ischemic neurovascular diseases. PMID- 19565182 TI - Activity-dependent survival of neurons in culture: a model of slow neurodegeneration. AB - Central neurons express persistent spontaneous electrical network activity both in the developing brain in vivo as well as in dissociated cultures. This electrical activity is important for the formation of connections among neurons, and for their survival. Prolonged suppression of the spontaneous activity using the sodium channel blocker tetrodotoxin (TTX) causes the death of the cultured neurons. In the present study, we investigated molecular mechanisms that may underlie the activity-suppressed slow degeneration of cortical neurons in culture. Already after 6-7 days of exposure to TTX, neurons begin to express apoptotic vacuoles and shrunken dendrites. Eventually, neurons activate p53, caspase-3 and BAX, hallmarks of neuronal apoptosis, before they die. This death is restricted to neurons, and no parallel process is seen in glial cells that co exist in the culture. These experiments may lead to a better understanding of slow neuronal death, akin to that found in neurodegenerative diseases of the brain. PMID- 19565183 TI - A possible paraneoplastic neuromyelitis optica associated with lung cancer. AB - Neuromyelitis optica is associated with antibodies to aquaporin-4 channels and has been classically defined as the occurrence of bilateral optic neuritis and transverse myelitis with no other neurological involvement. We describe here a 63 year-old woman who presented to our local hospital with an acute, bilateral visual loss followed by progressive paraparesis. Medical evaluations revealed a lung tumor and spinal MRI showed multiple cervical-dorsal hyperintense T(2) lesions with mild mass effect. Testing for NMO-Ig resulted positive. Patient's diagnosis was thus consistent with a possible paraneoplastic neuromyelitis optica associated with non-small cell lung cancer and evidence of NMO-Ig. PMID- 19565184 TI - S-adenosylmethionine and S-adenosylhomocysteine levels in the aging brain of APP/PS1 Alzheimer mice. AB - Hyperhomocysteinemia and factors of homocysteine metabolism, S adenosylhomocysteine (AdoHcy) and S-adenosylmethionine (AdoMet), may play a role in Alzheimer's disease (AD). With liquid-chromatography-tandem-mass-spectrometry AdoMet and AdoHcy were determined in brains of 8- and 15-month-old APP/PS1 Alzheimer mice, and their possible roles in AD brains investigated. The finding that AdoMet levels do not differ between the genotypes in (young) 8-month-old mice, but are different in (older) 15-month-old APP/PS1 mice compared to their wild-type littermates, suggests that alterations in AdoMet are a consequence of AD pathology rather than a cause. During aging, AdoMet levels decreased in the brains of wild-type mice, whereas AdoHcy levels diminished in both wild type and APP/PS1 mice. The finding that AdoMet levels in APP/PS1 mice are not decreased during aging (in contrast to wild-type mice), is probably related to less demand due to neurodegeneration. No effect of the omega-3 fatty acid docosahexaenoic acid (DHA) or cholesterol-enriched diets on AdoMet or AdoHcy levels were found. PMID- 19565185 TI - Intracranial and intraspinal hemorrhage following spinal anesthesia. AB - Spinal anesthesia (SA), accounting for more than 50% of regional anesthesias in the spinal region, is generally perceived as simple and safe. Our purpose is to increase awareness of hemorrhagic complications following SA. A 69-year-old male without either coagulation disorders or anticoagulant/antiplatelet therapy developed acute radiculopathy, and severe mental confusion after SA for prostatectomy. CT showed intracranial subarachnoid and intraventricular acute hemorrhage. Cerebral angiography was negative. MRI showed subarachnoid and subdural hematoma in the dorsolumbar spine. Seven-year follow-up showed permanent cognitive and radicular damage. Multiple attempts for SA most likely caused spinal vessels rupture, either directly or indirectly by inducing differential pressure changes between cerebrospinal fluid and intravascular spaces; however, definite mechanisms have not been completely understood. Patients undergoing spinal puncture must report any neurological abnormality, which may result in irreversible damage. Cases of altered consciousness require an extensive neuroradiological evaluation. Proper competency of physicians responsible for spinal puncture is mandatory. PMID- 19565187 TI - Phase II study of darinaparsin in patients with advanced hepatocellular carcinoma. AB - BACKGROUND: Darinaparsin is a novel organic arsenic that reaches higher intracellular concentration with decreased toxicity compared to inorganic arsenic. We conducted a multi-center phase II study with darinaparsin in patients with advanced HCC. METHODS: Eligibility criteria included unresectable or metastatic measurable HCC, up to two prior systemic treatments, ECOG performance status < or = 2, Child Pugh Class A or B and adequate organ functions. Darinaparsin was administered at 420 mg/m(2) intravenously, twice weekly at least 72 h apart for 3 weeks in a 4-week cycle. The primary end point was response rate. A Simon two-stage design was used. RESULTS: Among 15 patients in the first stage, no objective responses were observed. Two patients had stable disease. The median number of cycles on study per patient was 2 (1-6). The median progression free survival and overall survival were 55 days (95% confidence interval: 50-59) and 190 days (95% confidence interval: 93-227), respectively. No treatment related hospitalizations or deaths occurred. Treatment related grade 1-2 toxicities included nausea, vomiting (26.7% each), fatigue (20%), anorexia and diarrhea (13.3% each). Grade 3 anorexia, wheezing, agitation, abdominal pain and SGPT were observed in 1 patient each (6.7%). One patient experienced grade 4 hypoglycemia (6.7%). CONCLUSIONS: Darinaparsin could be safely administered with tolerable toxicity profiles, and no QTc prolongation in patients with advanced HCC. However, at this dose and schedule, it has shown no objective responses in HCC and this trial was terminated as planned after the first stage of efficacy analysis. PMID- 19565188 TI - Immobilization of poly (ethylene imine) on poly (L-lactide) promotes MG63 cell proliferation and function. AB - Poly (ethylene imine) (PEI) is a polycation widely used for DNA transfection to cells but also applied as primary polycation for layer-by-layer (LBL) assembly of polyelectrolytes. The aim of the present study was to investigate the effect of modification with PEI on the biocompatibility of poly (L-lactide) (PLLA) films. PEI with different molecular weight was immobilized on PLLA by either adsorption or covalent binding. Cell morphologies, immuno-fluorescence staining, cell proliferation by lactate dehydrogenase assay and cell differentiation by alkaline phosphatase assay were utilized to assess the biocompatibility of the modified PLLA using osteoblast cell line MG63. Results revealed that PEI modification remarkably improved cell adhesion, viability, proliferation and function compared with plain PLLA. Hence, PEI-modified PLLA is acceptable as transfection vehicle for engineering of bone and other tissues, or as primary layer to allow LBL assembly to generate biomimetic surface coatings. PMID- 19565186 TI - Plant aquaporin selectivity: where transport assays, computer simulations and physiology meet. AB - Plants contain a large number of aquaporins with different selectivity. These channels generally conduct water, but some additionally conduct NH(3), CO(2) and/or H(2)O(2). The experimental evidence and molecular basis for the transport of a given solute, the validation with molecular dynamics simulations and the physiological impact of the selectivity are reviewed here. The aromatic/arginine (ar/R) constriction is most important for solute selection, but the exact pore requirements for efficient conduction of small solutes remain difficult to predict. Yeast growth assays are valuable for screening substrate selectivity and are explicitly shown for hydrogen peroxide and methylamine, a transport analog of ammonia. Independent assays need to address the relevance of different substrates for each channel in its physiological context. This is emphasized by the fact that several plant NIP channels, which conduct several solutes, are specifically involved in the transport of metalloids, such as silicic acid, arsenite, or boric acid in planta. PMID- 19565190 TI - Stability of high cell density brewery fermentations during serial repitching. AB - The volumetric productivity of the beer fermentation process can be increased by using a higher pitching rate (i.e. higher inoculum size). However, the decreased yeast net growth observed in these high cell density brewery fermentations can adversely affect the physiological stability throughout subsequent yeast generations. Therefore, different O(2) conditions (wort aeration and yeast preoxygenation) were applied to high cell density fermentation and eight generations of fermentations were evaluated together with conventional fermentations. Freshly propagated high cell density populations adapted faster to the fermentative conditions than normal cell density populations. Preoxygenating the yeast was essential for the yeast physiological and beer flavor compound stability of high cell density fermentations during serial repitching. In contrast, the use of non-preoxygenated yeast resulted in inadequate growth which caused (1) insufficient yield of biomass to repitch all eight generations, (2) a 10% decrease in viability, (3) a moderate increase of yeast age, (4) and a dramatic increase of the unwanted flavor compounds acetaldehyde and total diacetyl during the sequence of fermentations. Therefore, to achieve sustainable high cell density fermentations throughout the economical valuable process of serial repitching, adequate yeast growth is essential. PMID- 19565189 TI - Isolation and characterization of a novel cDNA encoding methyl jasmonate responsive transcription factor TcAP2 from Taxus cuspidata. AB - A novel transcription factor, TcAP2, was isolated from Taxus cuspidata by yeast one-hybrid strategy. This factor interacts with jasmonate- and elicitor responsive element. Analysis of the deduced TcAP2 amino acid sequence revealed that TcAP2 contained a conserved AP2/ethylene-responsive element binding protein domain that consisted of 268 amino acids in a potential nuclear localization sequence. The factor of TcAP2 had a high homology, in its AP2 domain, to other AP2 family members. Based on phylogenetic analysis, it was different from other five DRE-binding proteins in their evolutionary relationship. The transcription of TcAP2 gene in yew accumulated primarily in young organs, such as young stems. Quantitative real-time RT-PCR analysis indicated that TcAP2 gene was inducible to express by treatments with methyl jasmonate plus salicylic acid, high salinity, and cold. This gene showed no response to either abscisic acid or drought treatment. PMID- 19565191 TI - Electrochemical selectivity enhancement by using monosuccinyl beta-cyclodextrin as a dopant for multi-wall carbon nanotube-modified glassy carbon electrode in simultaneous determination of quercetin and rutin. AB - Monosuccinyl beta-cyclodextrin (succinyl-beta-CD) was synthesized and the selectivity to quercetin and rutin of the succinyl-beta-CD-modified, multi-wall carbon nanotube (MWNT)-coated, glassy carbon electrode [(succinyl-beta-CD + MWNT)/GCE] was investigated. (1)H NMR and MALDI-MS data confirmed molecular structure of the synthesized succinyl-beta-CD. As a dopant in carboxylated MWNT modified electrode, succinyl-beta-CD clearly separated the peak potential (E(p)) of quercetin from that of rutin. The measured peak potential separation (DeltaE(p)) was 110 mV. More favorable complexation between succinyl-beta-CD and quercetin may enhance relative selectivity to quercetin of the (succinyl-beta-CD + MWNT)/GCE in quercetin-rutin mixture as compared to the beta-CD-modified GCE. PMID- 19565192 TI - Classification of FABP isoforms and tissues based on quantitative evaluation of transcript levels of these isoforms in various rat tissues. AB - In mammals, 10 isoforms of fatty acid-binding protein (FABP) are expressed in various tissues. To understand the role of multiple FABP isoforms, we have quantitatively examined the transcript levels of individual FABP isoforms in each of various tissues by Northern blotting using synthesized RNAs corresponding to the mRNA of each isoform as external standards. As a result, absolute transcript levels of individual FABP isoforms expressed in each tissue were successfully determined. The 10 FABP isoforms were classified into three categories: (1) isoforms FABP7 and 12 were not markedly expressed in any tissue examined; (2) isoforms showing certain transcript levels in multiple tissues; and (3) isoforms FABP6, 8, and 9, expressed at certain levels in one particular tissue. Based on the expression profiles of the isoforms, individual tissues were also classified into three groups: (1) tissues in which high-level expression of FABP isoforms was not observed, (2) tissues in which multiple FABP isoforms were expressed at certain levels, and (3) tissues in which a single FABP isoform was dominantly expressed at a certain level. These results give a better understanding of the meaning of the presence of multiple FABP isoforms in mammals. PMID- 19565193 TI - PCR-based cloning of the complete mouse mitochondrial genome and stable engineering in Escherichia coli. AB - We have devised a method for cloning an entire mammalian mitochondrial genome (mtDNA) in Escherichia coli using PCR-based amplification and sequential ligation. Here we test this approach by cloning the complete mouse mtDNA. The mtDNA was divided into four to five fragments based on unique restriction enzyme sites and amplified by high-fidelity long-range DNA polymerase. The synthesized fragments were cloned individually to test their toxicity in the E. coli host and then combined sequentially into a vector containing the E. coli R6K origin of DNA replication. The synthetic complete mouse mtDNA clones were replicated stably and faithfully in E. coli when maintained at moderately low copy numbers per cell. The sequence integrity of the synthetic mouse mtDNA clones was confirmed by nucleotide sequencing; no mutations or rearrangements in the genome were found. This approach can facilitate the cloning of entire mammalian mitochondrial genomes in E. coli and assist in the introduction of desired modifications into the mitochondrial genome. PMID- 19565194 TI - Superabsorbed alcohol dehydrogenase--a new catalyst for asymmetric reductions. AB - A new immobilisate of alcohol dehydrogenase (ADH) is described in which all components for the reaction, i.e. enzyme, the coenzyme NADP(+), the buffer and other cofactors (trace elements), are immobilized together. It is an all inclusive catalyst. The support is a cheap, commercially-available, superabsorbent polymer. The immobilisation is easy to achieve. The superabsorbed ADH is, even when dried, a stable and storable catalyst for at least five weeks at -18 degrees C. Asymmetric reductions of the prochiral ketones, acetophenone, 4 acetylpyridine and ethyl acetoacetate, with a superabsorbed ADH from Lactobacillus brevis (ADH 002) and a superabsorbed ADH from Thermoanaerobicum sp. (ADH 005) in 2-propanol as both the organic solvent and the cofactor-regenerating substrate are given. Yields of chiral (R) and (S)-alcohols from 97-100% were achieved within 18 to 48 h with enantiomeric excesses of >99%. The superabsorbed ADH was easily separated by filtration and could be reused at least four times. PMID- 19565198 TI - [Development of the Saxon Health Target "Active aging - aging in health, autonomy, and participation"]. AB - In Saxony, the consequences of demographic aging are observable already today. To manage the implications on the health sector, the Saxon Health Targets Steering Committee decided in March 2008 to develop a health target "Active Aging - Aging in Health, Autonomy, and Participation". Target development was based on a 7 level approach (fields of action, main goals, target areas, targets, strategies, intervention measures, indicators for evaluation). A quantitative content analysis was used to reveal 10 potential relevant fields of action, three of which were selected for target development. Targets were developed by 53 stakeholders in multiprofessional working groups. Criteria-based analyses were performed to assure appropriate scientific evidence and feasibility of targets and intervention measures. Over a period of 9 months, 24 targets were defined referring to the main goals "needs-based health care structures", "multiprofessional qualification", "self-rated health" and "intergenerational solidarity". Thirteen targets were developed into recommendations for specific intervention measures. Most of the proposed interventions aim to modify health related structures or psychosocial determinants of health in the elderly. The best recommendations for intervention measures shall be implemented in cooperation with interested decision-makers. PMID- 19565200 TI - [Dermatitis herpetiformis. An update of the pathogenesis]. AB - The multifactorial pathogenesis of dermatitis herpetiformis is reviewed in light of current experimental data. Genetic background, gluten consumption, and abnormal immune and autoimmune reactions are the most important pathogenetic factors, but other agents also participate in the disease development. The predisposing and inducing factors are summarized, while the pathophysiological steps leading to the development of skin symptoms are detailed. PMID- 19565201 TI - [Epidermolysis bullosa. An update]. AB - Epidermolysis bullosa (EB) represents a group of diseases characterized by skin fragility usually developing blisters after minimal trauma. The clinical picture ranges from mild subtypes with minor skin reactions to severe forms with lethal outcome within the first months of life. In the severe generalized subtypes, complications such as aggressive squamous cell carcinoma of the skin, anemia, esophageal stenosis and cardiomyopathy can occur so that multidisciplinary patient care is necessary. EB can be divided in four types--EB simplex (EBS), junctional EB (JEB), dystrophic EB (DEB) and Kindler syndrome. All together 33 subtypes can be distinguished. In 2008 a revised EB classification was introduced. Several eponyms for EB subtypes were replaced by descriptive names. The review presents the EB subtypes based on the new EB classification system, the molecular background and new therapeutic options. PMID- 19565202 TI - A genomic insight into diversity among tribal and nontribal population groups of Manipur, India. AB - Twenty autosomal markers, including linked markers at two gene markers, are used to understand the genomic similarity and diversity among three tribal (Paite, Thadou, and Kom) and one nontribal communities of Manipur (Northeast India). Two of the markers (CD4 and HB9) are monomorphic in Paite and one (the CD4 marker) in Kom. Data suggest the Meitei (nontribal groups) stand apart from the three tribal groups with respect to higher heterozygosity (0.366) and presence of the highest ancestor haplotypes of DRD2 markers (0.228); this is also supported by principal co-ordinate analysis. These populations are found to be genomically closer to the Chinese population than to other Indian populations. PMID- 19565203 TI - Apolipoprotein E polymorphism in hemodialyzed patients and healthy controls. AB - A possible association between end-stage renal disease (ESRD) and apolipoprotein E (APOE) polymorphism was found in some but not all studies. We have analyzed the APOE genotypes in 995 hemodialyzed patients (cases) and a sample of 6242 healthy individuals (controls) in the Czech Republic. There was a statistically significant difference in the frequency of APOE alleles between cases and controls, with more carriers of the APOE2 allele in ESRD patients (15.9%) than in controls (12.2%) (P = 0.005). The odds ratio of ESRD for the APOE2 allele, compared with APOE3E3 homozygotes, was 1.37 (95% confidence interval 1.13-1.67). The strength of the association increased with the time spent on hemodialysis: the odds ratio of all-cause ESRD in patients dialyzed for eight or more years was 1.27 (0.94-1.71), for 1-8 years 1.41 (1.09-1.81), and less than 1 year (nonsurvivors) 1.94 (0.88-4.18). This study suggests that the APOE2 allele is a possible genetic risk factor for all-cause ESRD in Caucasians. PMID- 19565204 TI - FAS and FAS ligand polymorphisms in the promoter regions and risk of gastric cancer in Southern China. AB - The FAS and FAS ligand (FASLG) system plays a key role in regulating apoptotic cell death, and corruption of this signaling pathway has been shown to participate in tumorigenesis. Functional promoter polymorphisms of the FAS and FASLG genes can alter transcriptional activities and thus alter risk of cancer. We hypothesized that the FAS -1377G>A, FAS -670A>G, and FASLG -844T>C polymorphisms in the promoter regions are associated with risk of gastric cancer. In a population-based case-control study of 332 gastric cancer cases and 324 controls, we genotyped these three polymorphisms and evaluated their association with risk of gastric cancer. We found that the FAS and FASL genotypes and the FAS haplotypes had no significant associations with risk of gastric cancer. In addition, there was no significant interaction between the FAS and FASL polymorphisms in the development of gastric cancer. The FAS and FASLG polymorphisms may not contribute to risk of gastric cancer in the southern Chinese population. PMID- 19565205 TI - Genetic stability of progeny from an artificial allotetraploid carp using sperm from five fish species. AB - Microsatellite markers and D-loop sequences of mtDNA from a female allotetraploid parent carp and her progenies of generations 1 and 2 induced by sperm of five distant fish species were analyzed. Eleven microsatellite markers were used to identify 48 alleles from the allotetraploid female. The same number of alleles (48) appeared in the first and second generations of the gynogenetic offspring, regardless of the source of the sperm used as an activator. The mtDNA D-loop analysis was performed on the female tetraploid parent, 25 gynogenetic offspring, and 5 sperm-donor species. Fourteen variable sites from the 1,018 bp sequences were observed in the offspring as compared to the female tetraploid parent. Results from D-loop sequence and microsatellite marker analysis showed exclusive maternal transmission, and no genetic information was derived from the father. Our study suggests that progenies of artificial tetraploid carp are genetically stable, which is important for genetic breeding of this tetraploid fish. PMID- 19565206 TI - [Sarcoidosis of lymph nodes in the submandibular compartment]. AB - Sarcoidosis is a granulomatous systemic disease of unknown etiology. Besides the landmark pulmonary lesions, extrathoracic manifestations of the disease can also occur. We report the case of a 53-year-old woman with an obscure swelling of both submandibular compartments. The radiological and pathohistological evaluations confirmed the uncommon diagnosis of sarcoidosis of the submandibular compartment. The tumor in each compartment consisted of a huge lymph node conglomerate respectively displacing the submandibular gland. The major salivary glands and the thorax were not involved. PMID- 19565207 TI - [Atypical gout tophi for differential diagnosis of head and neck tumors : case report and review of the literature]. AB - Gout is a mostly hereditary metabolic disease and is considered a disease of affluence. The disease is promoted by a purine-rich diet and shows an intermittent course of inflammatory joint manifestations and periods free of symptoms. The pathognomonic sign of the disease is an acute and very painful monarthritis with typical local deposits of uric acid, so-called gout tophi. No or inadequate treatment leads to the chronic form of gouty arthritis characterized more by joint destruction than by persistent pain. In head and neck gout tophi are seen as nodular lesions along the outer helical edges of the auricle. A case report of gout manifestation in the infratemporal fossa, deriving from the temporomandibular joint, with arrosion of the bony skull base demonstrates gout as a relevant disease for the ENT clinician. Potential diagnostic difficulties as well as recommendations for a therapeutic regimen of gouty lesions in such critical localizations will be reviewed. PMID- 19565208 TI - [Rendezvous technique for complete hypopharyngeal stenosis. Indications and variations of combined endoscopy]. AB - BACKGROUND: Hypopharyngeal stricture can result from a number of causes including chemoradiotherapy and esophagectomy and leads to inability to swallow with aspiration as well as permanent dependence on a gastrostomy tube. Antegrade dilatation or puncture and local mitomycin C application are often unsuccessful and many patients require extensive surgery. METHODS: We report three cases of total hypopharyngeal stenosis with different clinical histories. We present our experience using three modifications of the combined anterograde-retrograde endoscopic technique with resection of the stenosis by laser technique under diaphanoscopic control. RESULTS: In all cases the hypopharyngeal-esophageal passage was restored. No complications occurred as a result of the procedure. CONCLUSION: Combined direct hypopharyngoscopy with retrograde esophagoscopy represents a viable alternative to more extensive approaches for recanalization of selected obstructing hypopharyngeal stenoses when antegrade identification of the esophageal entrance fails. PMID- 19565209 TI - [Steroid-refractory graft-versus-host disease: extracorporeal irradiation of leucocytes induces immunotolerance]. AB - A 42 year-old woman develops steroid refractory graft-versus-host disease (GVHD) after second allogeneic stem cell transplantation for acute myelogenous leukemia with severe GVHD of her skin with blisters, severe GVHD of her gut with watery and bloody diarrhea and GVHD of her liver with cholestasis. In a further attempt to control GVHD extracorporeal photochemotherapy is administered. The treatment exposures peripheral mononuclear cells to photoactivated psoralen before they subsequently are given back to the patient. This approach apparently offers selective immune tolerance. PMID- 19565210 TI - [Thoracic aorta aneurysms]. AB - Thoracic aorta aneurysms are life-threatening diseases which can lead to rupture or dissection due to structural alterations. The exact etiology is still unclear but extended life-expectancy with arterial hypertension, positive family history for aneurysmic diseases, atherosclerosis, smoking and chronic obstructive lung disease are all considered to be risk factors. Nowadays, a diagnosis can often be made in the symptom-free stage. The risk of rupture varies between 46 and 74% for a diameter clearly greater than 5.5 cm and with a 2-year mortality rate of 70%. The 5-year survival rate with conservative treatment can be as low as 13-34% compared to 70-79% after optimal surgical treatment. Medical, operative and percutaneous interventional therapeutic strategies are basically available. PMID- 19565211 TI - Deviance partitioning of host factors affecting parasitization in the European brown hare (Lepus europaeus). AB - Deviance partitioning can provide new insights into the ecology of host-parasite interactions. We studied the host-related factors influencing parasite prevalence, abundance, and species richness in European brown hares (Lepus europaeus) from northern Spain. We defined three groups of explanatory variables: host environment, host population, and individual factors. We hypothesised that parasite infection rates and species richness were determined by different host related factors depending on the nature of the parasite (endo- or ectoparasite, direct or indirect life cycle). To assess the relative importance of these components, we used deviance partitioning, an innovative approach. The explained deviance (ED) was higher for parasite abundance models, followed by those of prevalence and then by species richness, suggesting that parasite abundance models may best describe the host factors influencing parasitization. Models for parasites with a direct life cycle yielded higher ED values than those for indirect life cycle ones. As a general trend, host individual factors explained the largest proportion of the ED, followed by host environmental factors and, finally, the interaction between host environmental and individual factors. Similar hierarchies were found for parasite prevalence, abundance, and species richness. Individual factors comprised the most relevant group of explanatory variables for both types of parasites. However, host environmental factors were also relevant in models for indirect life-cycle parasites. These findings are consistent with the idea of the host as the main habitat of the parasite; whereas, for indirect life-cycle parasites, transmission would be also modulated by environmental conditions. We suggest that parasitization can be used not only as an indicator of individual fitness but also as an indicator of environmental quality for the host. This research underlines the importance of monitoring parasite rates together with environmental, population, and host factors. PMID- 19565212 TI - Omitting breakfast and lunch after injection of different long-acting insulin preparations at bedtime: a prospective study in patients with type 2 diabetes. AB - AIMS/HYPOTHESIS: The aim of this prospective trial was to compare the effect of different long-acting insulin preparations injected at bedtime on glucose concentrations in patients with type 2 diabetes omitting breakfast and lunch the next day. METHODS: Twenty patients (ten women) with type 2 diabetes who were on an intensified insulin therapy participated. Mean (+/-SD) age was 63 +/- 10 years, diabetes duration 18 +/- 9 years, BMI 32.5 +/- 5 kg/m(2), and HbA(1c) 7.3 +/- 0.7%. Patients received neutral protamine Hagedorn (NPH) insulin, insulin detemir or insulin glargine for at least 2 months; doses were adjusted to achieve morning blood glucose levels of <7 mmol/l. At the end of the respective treatment period, the long-acting insulin was injected at bedtime (at 22:45 hours) as usual but patients refrained from breakfast and lunch the next day; glucose was measured by a continuous glucose monitoring system (CGMS). RESULTS: Comparable glucose target ranges were reached at midnight (5.8 to 6.1 mmol/l) and at 07:00 hours (6.7 to 6.9 mmol/l) with all three insulin preparations, using mean doses of 29 +/- 10 U (NPH insulin), 33 +/- 13 U (insulin detemir), and 32 +/- 12 U (insulin glargine). Glucose levels between midnight and 07:00 hours were not significantly different for the three insulin preparations. Symptomatic hypoglycaemia did not occur from 08:00 to 16:00 hours; glucose concentrations during this time were slightly lower with NPH insulin than with insulin detemir (p = 0.012) and insulin glargine (p = 0.049). CONCLUSIONS/INTERPRETATION: Following bedtime injection of NPH insulin or of the analogues insulin detemir or insulin glargine, fasting glucose <7 mmol/l was achieved in the morning, without subsequent hypoglycaemia when participants continued to fast during the day. PMID- 19565213 TI - Insulin-requiring diabetes in rural Ethiopia: should we reopen the case for malnutrition-related diabetes? AB - AIMS/HYPOTHESIS: We evaluated the incidence of insulin-requiring diabetes in a rural area of sub-Saharan Africa. METHODS: Health surveillance data from a chronic disease programme in two zones of Ethiopia, Gondar and Jimma, were studied. The two zones have a population of more than 5,000,000 people. RESULTS: In Gondar Zone (1995-2008) and Jimma Zone (2002-2008) 2,280 patients presented with diabetes, of whom 1,029 (45%) required insulin for glycaemic control at diagnosis. The annual incidence of insulin-requiring diabetes was 2.1 (95% CI 2.0 2.2) per 100,000 and was twice as high in men (2.9 per 100,000) as in women (1.4 per 100,000). In both sexes incidence rates peaked at the age of 25 to 29 years. Incidence rates in the urban areas of Gondar and Jimma were five times higher than in the surrounding rural areas. Patients with insulin-requiring diabetes from rural and urban areas had a very low BMI and most were subsistence farmers or unemployed. CONCLUSIONS/INTERPRETATION: The typical patient with diabetes in rural Ethiopia is an impoverished, young adult male with severe symptoms requiring insulin for glycaemic control. The low incidence rates in rural compared with urban areas suggest that many cases of this disease remain undiagnosed. The disease phenotype encountered in this area of Africa is very different from the classical type 1 diabetes seen in the West and most closely resembles previous descriptions of malnutrition-related diabetes, a category not recognised in the current WHO Diabetes Classification. We believe that the case for this condition should be reopened. PMID- 19565216 TI - [Routine interventions and rare cases: latest original articles and case studies in orthopedics]. PMID- 19565214 TI - Risk of malignancies in patients with diabetes treated with human insulin or insulin analogues: a cohort study. AB - AIMS/HYPOTHESIS: The aim of this cohort study was to investigate the risk of malignant neoplasms and mortality in patients with diabetes treated either with human insulin or with one of three insulin analogues. METHODS: Data were provided by the largest German statutory health insurance fund (time-frame: January 1998 to June 2005 inclusive), on patients without known malignant disease who had received first-time therapy for diabetes mellitus exclusively with human insulin, aspart, lispro or glargine. The primary outcome was the diagnosis of a malignant neoplasm. Data were analysed by multiple Cox regression models adjusting for potential confounders. RESULTS: A total of 127,031 patients were included, with a mean follow-up time of 1.63 (median 1.41, maximum 4.41) years. A positive association between cancer incidence and insulin dose was found for all insulin types. Because patients receiving combined therapy with insulin analogues and human insulin were excluded, the mean daily dose was much lower for glargine than for human insulin, and a slightly lower cancer incidence in the glargine group was found. After adjusting for dose, a dose-dependent increase in cancer risk was found for treatment with glargine compared with human insulin (p < 0.0001): the adjusted HR was 1.09 (95% CI 1.00 to 1.19) for a daily dose of 10 IU, 1.19 (95% CI 1.10 to 1.30) for a daily dose of 30 IU, and 1.31 (95% CI 1.20 to 1.42) for a daily dose of 50 IU. No increased risk was found for aspart (p = 0.30) or lispro (p = 0.96) compared with human insulin. CONCLUSIONS/INTERPRETATION: Considering the overall relationship between insulin dose and cancer, and the lower dose with glargine, the cancer incidence with glargine was higher than expected compared with human insulin. Our results based on observational data support safety concerns surrounding the mitogenic properties of glargine in diabetic patients. Prospective long-term studies are needed to further evaluate the safety of insulin analogues, especially glargine. PMID- 19565217 TI - Clinical relevance of static and dynamic tests after anatomical double-bundle ACL reconstruction. AB - A computer navigation system was used to collect kinematic data of 18 subjects undergoing ACL reconstruction. Surgical procedure was an anatomical four-tunnel hamstring double-bundle reconstruction. Static laxity and dynamic laxity were analyzed before and after graft passage and fixation. Correlations between static and dynamic laxities as well as with the preoperative IKDC score were determined. Static and dynamic laxities were significantly reduced after anatomic double bundle ACL reconstruction (P < 0.05). There were no significant correlations between static and dynamic laxities (r < 0.4, P > 0.05). The preoperative IKDC score was only related to preoperative dynamic laxity (P < 0.01). The dynamic evaluation of pivot shift is able to better describe knee laxity, in particular rotational laxities and has no correlation with static laxity. PMID- 19565219 TI - Development of new SLAP lesion after the arthroscopic, isolated decompression of ganglion cyst of the shoulder. AB - Ganglion cysts of the shoulder are rare, and the pathogenesis is similar to that of meniscal cysts. We present details of two cases of isolated ganglion cyst of shoulder which were treated arthroscopically. Both patients following 1 year after the surgery, complained of nonspecific shoulder pain and magnetic resonance imaging revealed new SLAP lesion which was treated arthroscopically. PMID- 19565218 TI - The femoral insertions of the anteromedial and posterolateral bundles of the anterior cruciate ligament: a radiographic evaluation. AB - The aim of this radiographic study was to visualize the femoral insertion sites of the anteromedial (AM) and posterolateral (PL) bundle of the anterior cruciate ligament (ACL) on lateral radiographs in different angles of knee flexion to gain better understanding for arthroscopic femoral tunnel placement in ACL double bundle reconstruction. Four fresh cadaveric knees with an intact ACL were dissected to isolate the AM and PL bundle of the ACL. We obtained lateral radiographs of each knee over the range of 0 degrees -90 degrees flexion in 30 degrees increments after painting the bundles with a radiopaque tantalum powder. The center of the radiographically marked femoral insertion was defined for each bundle on the lateral roentgenogram. We analyzed the relationship of knee flexion and the projection of the relative position of the femoral insertion sites of both bundles of the ACL on the lateral roentgenogram. The centre of the PL bundle visualized more anterior and distal than the centre of the AM bundle with the knee held in 90 degrees flexion. The centers of the AM and PL bundle were horizontally aligned when the knee was flexed over 90 degrees . The resulting images allow a radiographic description of the femoral insertion sites of both bundles in different angles of knee flexion. It is essential to be aware of the degree of knee flexion when drilling the femoral tunnels. PMID- 19565220 TI - Correlation between the rotational degree of the dial test and arthroscopic and physical findings in posterolateral rotatory instability. AB - Diagnosing posterolateral rotatory instability (PLRI) is difficult because it occurs rarely, takes time to manifest, and no single definitive tool exists in diagnosing posterolateral corner injuries. We sought to evaluate the correlation between rotational degrees in the dial test and physical and arthroscopic findings that surgically verified PLRI in the knee joint. Patients (n = 67) who were diagnosed as having PLRI and received posterolateral reconstruction (n = 57) and repair (n = 10) from 1998 to 2006 were recruited. Preoperative physical findings were evaluated under anesthesia, and arthroscopic findings during surgeries were analyzed. A dial test with post-anesthesia (spinal or general) was conducted. We divided patients into three subgroups (A: <15 degrees, B: 15-20 degrees, and C: >20 degrees), according to differences in rotational degrees in the dial test. All tests showed significant differences among the three groups and positive findings increased as the rotational degrees increased. No test showed a statistically significant difference in the specific group. All tests except for popliteal hiatus widening showed significant differences among the three groups and positive findings also increased as the rotational degrees increased. The total incidence and positive rate of each physical examination and the popliteal hiatus arthroscopic findings in PLRI significantly increased as the rotational degree in the dial test increased. The number of positive findings on physical examination was larger than the number of positive findings by arthroscopy in all three groups. PMID- 19565221 TI - Prognostic value of chondral defects on the outcome after arthroscopic treatment of acetabular labral tears. AB - Pathology of the acetabular labrum plays an increasing role in the treatment of hip pain. Hip arthroscopy has proven its clinical value as a useful procedure for successful treatment of labral tears. Until today, only a few studies have investigated the influence of articular cartilage defects on the clinical outcome of partial arthroscopic labrum resection in a larger patient population. We prospectively evaluated patients with an intraoperatively proven labral lesion/tear without any radiological and arthroscopical sign of a concomitant bony femoroacetabular impingement or hip dysplasia for a minimum postoperative follow-up of 2 years. Cartilage defects were classified according to Outerbridge and divided into two subgroups: Outerbridge < or = 1 and Outerbridge > or = 2, respectively. To evaluate combined results, various established scoring systems (visual analogue scale, modified Harris Hip Score, Larson Hip Score) were used. Out of 54 originally enrolled patients, 50 individuals (29 female, 21 male) with a median age of 33 years (range 15-49) were available for follow-up after a mean of 34 (range 24-48) months. At follow-up, the total study population experienced significant improvement in pain and in the combined evaluation scales (Larson Hip Score/MHHS). When patients were categorized into two subgroups, either with intraoperatively present or absent articular cartilage defects, our data indicated that subjects with no degenerative changes of the articular cartilage surface significantly improved in the applied clinically scoring systems. In contrast, in patients with an articular cartilage lesion during hip arthroscopy score values had a tendency to be unimproved or even deteriorated at follow-up. Regression analysis revealed a significant negative correlation between postoperative outcome and the grading of the coexistent articular cartilage defect. On the basis of our investigation, we conclude that partial arthroscopic resection of a torn labrum without attending bone deformity (dysplasia or femoroacetabular impingement) can reveal good and satisfied results. Depending on the extent of a coexisting articular cartilage defect subjective clinical results are compromised. PMID- 19565223 TI - Preparation and inhibition of interceptive actions. AB - Two experiments aimed to provide an estimate of the last moment at which visual information needs to be obtained in order for it to be used to initiate execution of an interceptive movement or to withhold execution of such a movement. In experiment 1, we sought to estimate the minimum time required to suppress the movement when the participants were first asked to intercept a moving target. In experiment 2, we sought to determine the minimum time required to initiate an interceptive movement when the participants were initially asked to keep stationary. Participants were trained to hit moving targets using movements of a pre-specified duration. This permitted an estimate of movement onset (MO) time. In both experiments the requirement to switch from one prepared course of action to the other was indicated by changing the colour of the moving target at times prior to the estimated MO. The results of the experiments showed that the decision to execute or suppress the interception must be made no less than about 200 ms before MO. PMID- 19565224 TI - Early nerve ending rescue from oxidative damage and energy failure by L: carnitine as post-treatment in two neurotoxic models in rat: recovery of antioxidant and reductive capacities. AB - Cell rescue is a primary need during acute and chronic insults to the central nervous system. Functional preservation during the early stages of toxicity in a given degenerative event may represent a significant amelioration of detrimental processes linked to neuronal cell loss. Excitotoxicity and depleted cellular energy are toxic events leading to cell death in several neurodegenerative disorders. In this work, the effects of the well-known antioxidant and energy precursor, L: -carnitine (L: -CAR), were tested as a post-treatment in two neurotoxic models under in vitro and in vivo conditions. The experimental models tested included: (1) a typical excitotoxic and pro-oxidant inducer, quinolinic acid (QUIN); and (2) a mitochondrial energy inhibitor, 3-nitropropionic acid (3 NP). For in vitro studies, increasing concentrations of L: -CAR (10-1,000 microM) were added to the isolated brain synaptosomes at different times (1, 3 and 6 h) after the incubation with toxins (100 microM QUIN and 1 mM 3-NP), and 30 min later, lipid peroxidation (LP) and mitochondrial dysfunction (MD) were evaluated. For in vivo purposes, L: -CAR (100 mg/kg, i.p.) was given to rats either as a single administration 120 min after the intrastriatal infusion of QUIN (240 nmol/microl) or 3-NP (500 nmol/microl), or for 7 consecutive days (starting 120 min post-lesion). LP and MD were evaluated 4 h and 7 days post-lesions in isolated striatal synaptosomes. Our results show that, despite some variations depending on the toxic model tested, the time of exposure, or the biomarker evaluated, nerve ending protection can be mostly achieved by L: -CAR within the first hours after the toxic insults started, suggesting that targeting the ongoing oxidative damage and/or energy depletion during the first stages of neurotoxic events is essential to rescue nerve endings. PMID- 19565225 TI - The impact of real and illusory target perturbations on manual aiming. AB - This experiment was designed to determine if real and illusory shifts in target position at movement initiation affect the same online corrective processes. Adult participants completed rapid goal-directed movements toward the vertex of a target "T" located at the midline, 25 cm distal to a small home position. At movement initiation, the target either stayed the same, shifted its real position, its illusory position or both. The real perturbation involved a 2.5 mm shift either toward or away from the body. For the illusory perturbation, the horizontal portion of the "T" changed to inward or outward Muller-Lyer wings. Both the real and the illusory perturbation affected movement outcome. The two manipulations began to have their impact at peak velocity. Because both perturbations affected mid to late trajectory control and because their effects were not independent, we concluded that real and illusory target shifts impact late visual motor control associated with a comparison between the position of the limb and the perceived position of the target. PMID- 19565226 TI - Interactions between new and pre-existing dynamics in bimanual movement control. AB - Motor skills are commonly acquired through practice. This process not only involves acquisition of the particular task demands but also requires overcoming pre-existing modes. In the present study, interactions between new and intrinsic dynamics were evaluated. Accordingly, bimanual finger tapping with a 2:1 ratio was performed according to two training schedules: continuous (consecutive trials) and interrupted (non-consecutive trials with intermediate 1:1 in-phase performances). In addition, in-phase and anti-phase were probed before and after training. Behavioral output was assessed by means of temporal accuracy and variability, whereas neural activation patterns were determined by EEG coherence. Results showed that continuous practice resulted in improved performance with reduced coherence across the motor network. For interrupted practice, behavioral execution ameliorated, although it was inferior to performance with continuous practice. In terms of neural changes, the degree of intrahemispheric and midline connectivity did not reduce with interrupted practice, whereas interhemispheric connectivity increased. This signifies that short-term motor consolidation of the 2:1 task was disrupted due to intermediate performance of the in-phase mode. Furthermore, the probed in-phase and anti-phase pattern showed no behavioral changes, although neural alterations occurred that depended on training schedule and coordination mode. Overall, the observations illustrate bidirectional interactions between new and inherent dynamics during motor acquisition, raising issues about effective methods for learning skills and scheduling of practices in neurorehabilitation. PMID- 19565227 TI - Diversity of grip in Macaca mulatta. AB - Much of the research on the neuronal basis of prehension focuses on macaque monkeys. Yet most of the behavioral description of grip types pertains to humans and apes. The purpose of the present study was to provide a catalogue and description of basic grip behavior in macaque monkeys. The observational study explored the diversity of grasping behavior in 157 semi-free ranging rhesus macaques. Video footage of monkeys grasping objects ad libitum was analyzed frame by-frame, and grips were classified based on the skin surface areas that contacted the object. When monkeys held objects for manipulation, 15 distinct grip categories were observed. When monkeys held support points during climbing, two grip categories were observed. Not all grips were performed with the hand. Some involved the mouth, the foot, or an opposition between the forearm and chest. Grip in macaque monkeys is more diverse than the narrow range of grip that is typically studied. PMID- 19565228 TI - Neonatal exposure to MK801 promotes prepulse-induced delay in startle response time in adult rats. AB - The acoustic startle reflex in rats can be inhibited if a prepulse stimulus is presented just before the startle stimulus (prepulse inhibition; PPI). When postnatal day 7 (P7) rats are exposed to agents that block the NMDA receptor (NMDAR), robust apoptosis is observed within hours and is thought to be followed at later ages by a significant loss of PPI. To understand these observations further, we exposed rat pups to vehicle or the NMDAR antagonist MK801 (1 mg/kg) at P6, P8, and P10. We then examined animals for PPI at P28 and P56. Compared to vehicle controls, we found no evidence for PPI deficits in the MK801-treated group, although we did observe prepulse-induced delay in response time at P56 (but not at P28). In a parallel study, we also performed histological analysis of brain sections for evidence of the pro-apoptotic marker activated caspase-3, 8 h after vehicle or MK801 injection into P6 animals. We found that there was a robust increase in this marker of cell death in the inferior colliculus of MK801 compared to vehicle-treated animals. Thus, transient blockade of the NMDAR during the postnatal period not only promotes early apoptosis in a brain region critical for acoustic processing but also leads to auditory deficits at a later age, suggesting that injury-induced loss of collicular neurons leads to network reorganization in the auditory system that is progressive in nature. PMID- 19565229 TI - Sensory information in perceptual-motor sequence learning: visual and/or tactile stimuli. AB - Sequence learning in serial reaction time (SRT) tasks has been investigated mostly with unimodal stimulus presentation. This approach disregards the possibility that sequence acquisition may be guided by multiple sources of sensory information simultaneously. In the current study we trained participants in a SRT task with visual only, tactile only, or bimodal (visual and tactile) stimulus presentation. Sequence performance for the bimodal and visual only training groups was similar, while both performed better than the tactile only training group. In a subsequent transfer phase, participants from all three training groups were tested in conditions with visual, tactile, and bimodal stimulus presentation. Sequence performance between the visual only and bimodal training groups again was highly similar across these identical stimulus conditions, indicating that the addition of tactile stimuli did not benefit the bimodal training group. Additionally, comparing across identical stimulus conditions in the transfer phase showed that the lesser sequence performance from the tactile only group during training probably did not reflect a difference in sequence learning but rather just a difference in expression of the sequence knowledge. PMID- 19565230 TI - Reflexive social attention is mapped according to effector-specific reference systems. AB - Studies exploring reflexive joint attention report that attention is more powerfully captured by interfering social signals (such as others' gaze or hand orientation) than by non-biological directional cues (such as an arrow). However, what remains unknown is whether these effects are mapped in purely spatial or in body-part specific reference frames. Changes of a central, black fixation point into blue or orange were the imperative instruction signal for the experimental subjects to make a leftward or a rightward movement (saccades in Study 1 and hand pointing in Study 2) while ignoring distracting stimuli (leftward or rightward oriented gaze, hand pointing or arrow). Gaze and pointing hand distracters that were directionally incongruent with the instruction cue impaired the goal-driven saccadic and pointing performance, respectively. This pattern of results indicates that reflexive social attention is mapped not only in spatial but also in body-part specific reference frames. PMID- 19565231 TI - Relationships between motor unit size and recruitment threshold in older adults: implications for size principle. AB - As a part of the aging process, motor unit reorganization occurs in which small motoneurons reinnervate predominantly fast-twitch muscle fibers that have lost their innervation. We examined the relationship between motor unit size and the threshold force for recruitment in two muscles to determine whether older individuals might develop an alternative pattern of motor unit activation. Young and older adults performed isometric contractions ranging from 0 to 50% of maximal voluntary contraction in both the first dorsal interosseous (FDI) and tibialis anterior (TA) muscles. Muscle fiber action potentials were recorded with an intramuscular needle electrode and motor unit size was computed using spike triggered averaging of the global EMG signal (macro EMG), which was also obtained from the intramuscular needle electrode. As expected, older individuals exhibited larger motor units than young subjects in both the FDI and the TA. However, moderately strong correlations were obtained for the macro EMG amplitude versus recruitment threshold relationship in both the young and older adults within both muscles, suggesting that the size principle of motor unit recruitment seems to be preserved in older adults. PMID- 19565233 TI - Follow-up skeletal surveys for nonaccidental trauma: can a more limited survey be performed? AB - BACKGROUND: Studies have demonstrated the value of the follow-up skeletal survey in identifying additional fractures, clarifying indeterminate findings, and improving dating of skeletal injuries in victims of physical abuse. OBJECTIVE: To determine whether a more limited follow-up survey could yield the same radiologic data as a full follow-up survey. MATERIALS AND METHODS: The study cohort comprised 101 children who had follow-up surveys that met our inclusion criteria. Consensus readings of both original and follow-up surveys were performed by two pediatric radiologists. These results were compared to determine additional findings from the follow-up surveys. Limited skeletal survey protocols were evaluated to determine whether they would detect the same fractures seen with a complete osseous survey. RESULTS: In the 101 children 244 fractures were identified on the initial osseous survey. Follow-up surveys demonstrated new information in 38 children (37.6%). A 15-view limited follow-up survey identified all additional information seen on the complete follow-up survey. CONCLUSION: Our data demonstrate that a 15-view limited follow-up skeletal survey could be performed without missing clinically significant new fractures and still allow proper identification of confirmed fractures or normal findings. A limited survey would decrease radiation dose in children. PMID- 19565234 TI - Response to Galaznik, Cohen & Scheimberg, and Rorke-Adams & Christian. PMID- 19565235 TI - Imaging recommendations in paediatric uroradiology. Minutes of the ESPR uroradiology task force session on childhood obstructive uropathy, high-grade fetal hydronephrosis, childhood haematuria, and urolithiasis in childhood. ESPR Annual Congress, Edinburgh, UK, June 2008. PMID- 19565232 TI - Administration of sesamol improved blood-brain barrier function in streptozotocin induced diabetic rats. AB - Uncontrolled or poorly controlled blood glucose during diabetes is an important factor in worsened vascular function. While evidence suggests that hyperglycemia induced oxidative stress plays a prominent role in development of microangiopathy of the retina, kidney, and nerves, the role oxidative stress plays on blood-brain barrier (BBB) function and structure has lagged behind. In this study, a natural antioxidant, sesamol, was administered to streptozotocin (STZ)-induced diabetic rats to examine the role that oxidative stress plays on BBB structure and function. Experiments were conducted at 56 days after STZ injection. Male Sprague Dawley rats randomly were divided into four treatment groups CON--control; STZ- STZ-induced diabetes; CON + S--control + sesamol; STZ + S--STZ-induced diabetes + sesamol. Functional and structural changes to the BBB were measured by in situ brain perfusion and western blot analysis of changes in tight junction protein expression. Oxidative stress markers were visualized by fluorescent confocal microscopy and assayed by spectrophotometric analysis. Results demonstrated that the increased BBB permeability observed in STZ-induced diabetic rats was attenuated in STZ + S rats to levels observed in CON. Sesamol treatment reduced the negative impact of STZ-induced diabetes on tight junction protein expression in isolated cerebral microvessels. Oxidative stress markers were elevated in STZ as compared to CON. STZ + S displayed an improved antioxidant capacity which led to a reduced expression of superoxide and peroxynitrite and reduced lipid peroxidation. In conclusion, this study showed that sesamol treatment enhanced antioxidant capacity of the diabetic brain and led to decreased perturbation of hyperglycemia-induced changes in BBB structure and function. PMID- 19565236 TI - Secreted production of an elastin-like polypeptide by Pichia pastoris. AB - Elastin-like polypeptides (ELPs) are biocompatible designer polypeptides with inverse temperature transition behavior in solution. They have a wide variety of possible applications and a potential medical importance. Currently, production of ELPs is done at lab scale in Escherichia coli shake flask cultures. With a view to future large scale production, we demonstrate secreted production of ELPs in methanol-induced fed-batch cultures of Pichia pastoris and purification directly from the culture medium. The production of ELPs by P. pastoris proved to be pH dependent within the experimental pH range of pH 3 to 7, as an increasing yield was found in cultures grown at higher pH. Because ELP produced at pH 7 was partly degraded, a pH optimum for production of ELP was found at pH 6 with a yield of 255 mg of purified intact ELP per liter of cell-free medium. PMID- 19565237 TI - Improved production of various polyunsaturated fatty acids through filamentous fungus Mortierella alpina breeding. AB - Studies on the application of functional lipids such as polyunsaturated fatty acids (PUFAs) have proceeded in various fields regarding health and dietary requirements in a search for novel and rich sources. Filamentous fungus Mortierella alpina 1S-4 produces triacylglycerols rich in arachidonic acid, ones reaching 20 g/L and containing 30-70% arachidonic acid as to the total fatty acids. Mutants derived from M. alpina 1S-4, defective in Delta5 and Delta6 desaturases, accumulate triacylglycerols rich in unique PUFAs, i.e., dihomo-gamma linolenic acid and Mead acid, respectively. Furthermore, various mutants derived from M. alpina 1S-4 have led to the production of oils containing n-1, n-3, n-4, n-6, n-7, and n-9 PUFAs. A variety of genes encoding fatty acid desaturases and elongases involved in PUFA biosynthesis in M. alpina 1S-4 has been isolated and characterized. Molecular breeding of M. alpina strains by means of manipulation of these genes facilitates improvement of PUFA productivity and elucidation of the functions of enzymes involved in PUFA biosynthesis. PMID- 19565238 TI - Cam-type femoral-acetabular impingement: is the alpha angle the best MR arthrography has to offer? AB - INTRODUCTION: In our institutional experience, determination of the alpha (alpha) angle at MR arthrography as an indicator of the likelihood of cam-type femoroacetabular impingement (FAI) is fraught with inconsistency. The aims of this study were to quantify the degree of variability in and calculate the diagnostic accuracy of the alpha angle in suggesting a diagnosis of cam impingement, to determine the accuracy of a positive clinical impingement test, and to suggest alternative MR arthrographic measures of femoral head-neck overgrowth and determine their diagnostic utilities. MATERIALS AND METHODS: We carried out a retrospective analysis of MR arthrographic studies performed during a 4-year period, combined with chart analysis, which allowed identification of 78 patients in whom surgical correlation was also available. The status of a preoperative clinical impingement test was also noted. Patients were designated as having cam-type FAI (Group A, n = 39) if intra-operative femoral head-neck junction bony osteochondroplasty/arthoscopic femoral debridement was performed. Group B (n = 39) acted as controls. Three radiologists independently and blindly performed a series of measurements (alpha angle and two newly proposed measurements) in each patient on two separate occasions. An alpha angle of greater than 55 degrees was considered indicative of the presence of cam-type FAI. RESULTS: Performance values for alpha angle measurement were poor for each observer. There was considerable (up to 30% of the mean value) intra-observer variability between the first and second alpha angle measurements for each subject. Binary logistic regression analysis confirmed that the alpha angle is of no value in predicting the presence or absence of cam-FAI. A statistically significant difference existed between Groups A and B with regard to the newly proposed anterior femoral distance (AFD; p = 0.004). Using an AFD value of 3.60 mm or greater as being indicative of the presence of cam-FAI yields a 0.67 performance measure (95% confidence interval 0.55-0.79). The second proposed parameter (femoral neck ratio) was of no value in suggesting the presence or absence of this condition. The sensitivity, specificity, and positive and negative predictive values of the clinical impingement test were 76.9%, 87.2%, 85.7% and 79.1% respectively. CONCLUSIONS: Femoral alpha angle measurement is associated with considerable variability. This index performed poorly in our patient population and was statistically of no value in suggesting the presence or absence of cam-FAI. One of our proposed measures, the AFD, outperformed the alpha angle, though to an insufficient degree to suggest its routine incorporation into clinical practice. Our experience suggests that the clinical impingement test remains the most reliable predictor of the presence of this condition. PMID- 19565240 TI - Comparative evaluations of norepinephrine transporter radioligands with reference tissue models in rhesus monkeys: (S,S)-[18F]FMeNER-D2 and (S,S)-[11C]MeNER. AB - PURPOSE: Although two norepinephrine transporter (NET) radioligands, (S,S) [(18)F]FMeNER-D(2) and (S,S)-[(11)C]MeNER, have been reported to have different characteristics, there has not been a direct comparative study in vivo. METHODS: In this preliminary study, two rhesus monkeys underwent three PET measurements each, one with (S,S)-[(18)F]FMeNER-D(2) and two with different radioactivity of (S,S)-[(11)C]MeNER (360 and 175 MBq). The regional brain uptakes and BP(ND) values from reference tissue models with different time lengths were compared. RESULTS: The regional uptakes in the thalamus and the brainstem with (S,S) [(11)C]MeNER did not always reach the peak during 123-min measurements while those with (S,S)-[(18)F]FMeNER-D(2) reached the peak before 30 min. The regional BP(ND) values of (S,S)-[(18)F]FMeNER-D(2) were stable after 150-min measurement lengths while those of (S,S)-[(11)C]MeNER were not stable during 123-min measurements. CONCLUSION: (S,S)-[(18)F]FMeNER-D(2) appears to be superior for the quantitative analysis with reference tissue models in the rhesus monkey. PMID- 19565241 TI - Predictors of hematological abnormalities in patients with chronic hepatitis C treated with interferon and ribavirin. AB - Hematological abnormalities including neutropenia, anemia, and thrombocytopenia are commonly seen in patients with chronic hepatitis C treated with pegylated interferon and ribavirin. The aim of this study was to identify factors which would help to predict the development of hematological abnormalities in patients with chronic hepatitis C treated with pegylated interferon and ribavirin. During a 4-year period, all patients with chronic hepatitis C started on treatment with pegylated interferon and ribavirin were identified. Patients were defined as having hematological abnormalities if they had the presence of either anemia, neutropenia, thrombocytopenia, or a combination of the above during treatment with pegylated interferon and ribavirin. A total of 136 patients with chronic hepatitis C were included in this study. Fifty-two (38.2%) of the patients developed significant hematological abnormalities during treatment with pegylated interferon and ribavirin with 28 (20.6%), 30 (22.1%), and 11 (8.1%) developed neutropenia, anemia, and thrombocytopenia, respectively. Genotype 1, history of hypertension, low baseline platelet count, low baseline hemoglobin, as well as a raised creatinine were significant factors associated with the development of hematological abnormalities. Significant hematological abnormalities are commonly present in patients with chronic hepatitis C treated with pegylated interferon and ribavirin. This study identifies pretreatment parameters that may help identify high-risk patients who are more likely to develop hematological abnormalities during treatment for chronic hepatitis C. PMID- 19565242 TI - [Intrahepatic sarcoma of the follicular dendritic cells]. AB - We report an intrahepatic sarcoma of the follicular dendritic cells in a 76-year old woman with a medical history of a hyaline-vascular type of Castleman's disease. We discuss the clinico-pathological findings, the pathogenesis and the differential diagnosis of this rare tumour entity. PMID- 19565239 TI - Molecular imaging of hypoxia with radiolabelled agents. AB - Tissue hypoxia results from an inadequate supply of oxygen (O2) that compromises biological functions. Structural and functional abnormalities of the tumour vasculature together with altered diffusion conditions inside the tumour seem to be the main causes of tumour hypoxia. Evidence from experimental and clinical studies points to a role for tumour hypoxia in tumour propagation, resistance to therapy and malignant progression. This has led to the development of assays for the detection of hypoxia in patients in order to predict outcome and identify patients with a worse prognosis and/or patients that would benefit from appropriate treatments. A variety of invasive and non-invasive approaches have been developed to measure tumour oxygenation including oxygen-sensitive electrodes and hypoxia marker techniques using various labels that can be detected by different methods such as positron emission tomography (PET), single photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), autoradiography and immunohistochemistry. This review aims to give a detailed overview of non-invasive molecular imaging modalities with radiolabelled PET and SPECT tracers that are available to measure tumour hypoxia. PMID- 19565243 TI - [Smoking and its effects on health : What the pathologist Schairer found already 70 years ago]. PMID- 19565245 TI - Cardiac spiral dual-source CT with high pitch: a feasibility study. AB - Increase of pitch in spiral CT decreases data acquisition time; dual-source CT (DSCT) systems provide improved temporal resolution. We evaluated the combination of these two features. Measurements were performed using a commercial DSCT system equipped with prototype software allowing pitch factors from p = 0.35 to 3.0. We measured slice sensitivity profiles as a function of pitch to assess spatial resolution in the z-direction and the contrast of structures moved periodically to measure temporal resolution. Additionally we derived modulation transfer functions to provide objective parameters; both spatial and temporal resolution were essentially unchanged even at high pitch. CT of the cardiac region of three pigs was performed at p = 3.0. In vivo CT images confirmed good image quality; direct comparison with standard low-pitch phase-correlated CT image datasets showed no significant difference. For a normalized z-axis acquisition of 12 cm, the corresponding effective dose value was 2.0 mSv for the high-pitch CT protocol. We conclude that spiral DSCT imaging with a pitch of 3.0 can provide unimpaired image quality with respect to spatial and temporal resolution. Applications to cardiac and thoracic imaging with effective dose below 1 mSv are possible. PMID- 19565244 TI - Isolation of DNA-methyltransferase genes from strawberry (Fragaria x ananassa Duch.) and their expression in relation to micropropagation. AB - DNA methylation can control gene expression and may also play a role in plant development. Methylation of cytosine residues in DNA is enzymatically catalyzed by DNA methyltransferases. In this study, full-length genomic genes and cDNAs of methyltransferase (MET1) and domain-rearranged methyltransferase (DRM) were isolated from strawberry (Fragaria x ananassa Duch.). Two genomic clones (FaMET1a and FaMET1b) encoding MET1 had open-reading frame of 4,695 and 4,671 nucleotides with two introns, respectively. Amino acid sequence comparison indicated high similarity (98.72% identity) of strawberry MET1 protein to other plant MET1 sequences. The full-length cDNA of strawberry DRM genes (FaDRMa, FaDRMb and FaDRMc) were 2,273, 2,282 and 2,288 bp, respectively. Ten introns with different sizes were dispersed in FaDRM genes. Similarly, FaDRMa, FaDRMb and FaDRMc had high-sequence similarity overall. Expressions of strawberry MET1 and DRM genes were compared among in vitro-micropropagated plants, generations of micropropagated plants and conventionally propagated plants. The transcriptional expressions of both FaMET1 and FaDRM genes were downregulated in micropropagated plants, and they were recovered in the first and second runner generations of micropropagated plants. However, there was a slighter difference in global DNA methylation rates between micropropagated plants and conventionally propagated plants. Therefore, there was no positive relation between global DNA methylation rates and the expression levels of MET1 and DRM genes. PMID- 19565246 TI - Can galactography-guided stereotactic, 11-gauge, vacuum-assisted breast biopsy of intraductal lesions serve as an alternative to surgical biopsy? AB - The purpose of this study was to determine the value of galactography-guided, stereotactic, vacuum-assisted breast biopsy (VABB) for the assessment of intraductal breast lesions and its potential as a therapeutic tool that could eliminate the need for surgical excision. Eighteen patients (median age 64 years, range 37-80) with nipple discharge and galactography-verified intraductal lesions underwent galactography-guided, stereotactic, 11-gauge VABB followed by surgery. Histopathology findings from VABB and subsequent surgery were compared. Underestimation and false-negative rates were assessed. After VABB, histopathology revealed invasive ductal carcinoma (IDC) in three (17%), ductal carcinoma in situ (DCIS) in six (33%), high-risk lesions in six (33%) and benign lesions in three (17%) cases. After surgical biopsy, histopathology confirmed the previously established diagnosis in 11 lesions (61%). The underestimation rate for high-risk lesions and DCIS was 50% (6/12). The false-negative rate was 7% (1/14). Histopathology examination after surgery showed that not a single lesion had been completely removed at VABB. Galactography-guided VABB is a feasible diagnostic tool. However, its value as a therapeutic procedure is limited because of the high number of underestimated and missed lesions and because of the histopathological detection of lesions' remnants in every case. Surgical excision should be the therapeutic gold standard in cases of pathological nipple discharge and galactography abnormalities. PMID- 19565247 TI - Perioperative analgesia strategies in fast-track pediatric surgery of the kidney and renal pelvis: lessons learned. AB - PURPOSE: Effective analgesia is essential for the success of fast-track (FT) pediatric surgery. Aim of the study was to achieve an optimal analgesia protocol for a comfortable postoperative course and early mobilization in children undergoing urological procedures. METHODS: A prospective study of two analgesia concepts in a series of children undergoing laparoscopic pyeloplasty (LP) and nephrectomy (LN) was performed. Thirty-six consecutive patients received analgesia according to the protocol "Fast Track I" (FT I), 23 patients according to the modified analgesia protocol "Fast Track II" (FT II). Differences between the protocols were frequency of pain measurement, more frequent use of local anesthesia and higher dosage of Acetaminophen. End points were pain intensity, use of opioids and side effects. Data derived from the German reimbursement system (G-DRG) regarding lengths of hospital stay were compared. RESULTS: On the day of operation (6:00 p.m.), pain scores of patients using FT II were significantly lower than those using the FT I protocol (1.51 + or - 0.4 FT II versus 3.8 + or - 0.4 FT I, p = 0.001). Pain scores still tended to be lower in the FT II group on the first postoperative day (8:00 a.m.), but from that point on, the difference was not significant (1.52 + or - 0.5 FT II versus 2.3 + or - 0.4 FT I, p = 0.186). Hospital stay was short, compared to the German average, in both groups. CONCLUSION: The modified analgesia protocol FT II leads to earlier and more effective pain reduction, lower use of opioids and fewer side effects than the previously used protocol FT I. Hence, modification of analgesia in pediatric urology is essential. PMID- 19565248 TI - Electrogenic Cl(-) secretion does not occur in the ileum of the Australian common brushtail possum, Trichosurus vulpecula, due to low levels of expression of the NaK2Cl cotransporter, NKCC1. AB - The colon of the brushtail possum does not have an electrogenic secretory response. Given the functional significance of electrogenic Cl(-) secretion in the intestine of eutherian mammals, we have investigated the secretory response in the small intestine of this marsupial. In the Ussing chamber cAMP-dependent secretagogues stimulated a sustained increase in ileal short-circuit current (Isc), whereas Ca(2+)-dependent secretagogues induced a transient increase. Both the responses were inhibited by mucosal addition of the anion channel blocker 5 nitro-2-(3-phenylpropylamino) benzoic acid (100 mciromol l(-1)), consistent with an anion secretory response. However, the responses were not inhibited by serosal bumetanide (10 mciromol l(-1)) and were independent of bath Cl(-), indicating that the stimulated ileal Isc does not involve electrogenic Cl(-) secretion driven by the NaK2Cl cotransporter, NKCC1. Consistent with this, there were low levels of NKCC1 expression in the ileal epithelium. In particular, NKCC1 expression in the ileal crypt cells was comparable to that of the villous cells. This differs from eutherian mammals where high levels of NKCC1 expression in the ileal crypt cells are associated with their role in Cl(-) secretion. The cAMP- and Ca(2+)-dependent secretory responses were inhibited by the removal of HCO(3) (-) suggesting that these responses were due to electrogenic HCO(3) (-) secretion. We conclude that the ileum of the possum does not secrete Cl(-) due to low levels of NKCC1 expression. It does however appear to secrete HCO(3) (-). These results are further significant examples of differences in the transport function of the possum intestinal epithelium compared with eutherian mammals. PMID- 19565249 TI - Development of myoglobin concentration and acid buffering capacity in harp (Pagophilus groenlandicus) and hooded (Cystophora cristata) seals from birth to maturity. AB - Pinnipeds rely on muscle oxygen stores to help support aerobic diving, therefore muscle maturation may influence the behavioral ecology of young pinnipeds. To investigate the pattern of muscle development, myoglobin concentration ([Mb]) and acid buffering ability (beta) was measured in ten muscles from 23 harp and 40 hooded seals of various ages. Adult [Mb] ranged from 28-97 to 35-104 mg g tissue( 1) in harp and hooded seals, respectively, with values increasing from the cervical, non-swimming muscles to the main swimming muscles of the lumbar region. Neonatal and weaned pup muscles exhibited lower (approximately 30% adult values) and less variable [Mb] across the body than adults. In contrast, adult beta showed little regional variation (60-90 slykes), while high pup values (approximately 75% adult values) indicate significant in utero development. These findings suggest that intra-uterine conditions are sufficiently hypoxic to stimulate prenatal beta development, but that [Mb] development requires additional postnatal signal such as exercise, and/or growth factors. However, because of limited development in both beta and [Mb] during the nursing period, pups are weaned with muscles with lower aerobic and anaerobic capacities than those of adults. PMID- 19565250 TI - Rivaroxaban and dabigatran etexilate: two new oral anticoagulants for extended postoperative prevention of venous thromboembolism after elective total hip arthroplasty. AB - Extended thromboprophylaxis is vital in patients undergoing total hip arthroplasty (THA) because of the prolonged risk of venous thromboembolism (VTE). Despite evidence that extended prophylaxis can reduce the incidence of symptomatic VTE in this high-risk patient population and the evidence-based guideline recommendations, a large proportion of patients still do not receive an adequate duration of thromboprophylaxis. This is partly due to the limitations of conventional anticoagulants, such as the subcutaneous route of administration or the requirement for routine coagulation monitoring and dose adjustment. New oral anticoagulants (such as the direct thrombin inhibitor dabigatran etexilate and the Factor Xa inhibitor rivaroxaban) could address the current unmet need. Phase III clinical studies in VTE prevention in patients undergoing THA and total knee arthroplasty (TKA) showed that dabigatran etexilate was non-inferior to the EU regimen of enoxaparin, but did not achieve non-inferiority to the US regimen of enoxaparin. In contrast, rivaroxaban demonstrated superiority to both enoxaparin regimens for the prevention of VTE after THA and TKA, without a significant increase in major bleeding rates. Their convenient, once-daily, fixed dosing, with no need for routine coagulation monitoring, could facilitate adherence to evidence-based guideline recommendations of extended thromboprophylaxis after THA. PMID- 19565251 TI - Epidural abscess associated with pyogenic spondylodiscitis of the lumbar spine; evaluation of a new MRI staging classification and imaging findings as indicators of surgical management: a retrospective study of 37 patients. AB - INTRODUCTION: The aim of this study was to review the patients with lumbar epidural abscess in terms of neurological morbidity, therapeutic outcome, and prognosis, while assessing the usefulness of a new MRI staging classification and specific imaging findings as indicators for surgical management. MATERIALS AND METHODS: We reviewed 37 patients who sustained epidural abscess associated with pyogenic spondylodiscitis of the lumbar spine. Ten patients were treated conservatively, while 27 required urgent or elective surgical drainage. We studied patients with respect to symptomatology, Frankel-American Spinal Injury Association (ASIA) scale evaluation and a new proposed system of MRI staging of pyogenic spondylodiscitis (stages I-V). RESULTS: Of the 37 patients with stage IV and V MRI lesions, 13 (35%) had septicemia and 8 (22%) presented with Frankel ASIA scale C-D neurological status. All cases with ringlike enhancement on gadolinium-enhanced MRI in the epidural abscess lesions were treated surgically. Progression of local kyphosis and loss of intervertebral disk height were significantly prevented in the surgical group (P < 0.05). Improvements of neurological status and laboratory data were better in the surgical group than the conservative group (P < 0.05), with significantly short hospital stay (P < 0.05). DISCUSSION: Epidural abscess associated with pyogenic spondylodiscitis presents with various neurological symptoms. In addition to assessment of progression by clinical symptomatology, modified neurological Frankel-ASIA scaling and the currently proposed MRI staging regimen may help to consider the timing of surgical intervention. In the acute, subacute or acute-on-chronic phase and the ringlike enhancement pattern of epidural abscess on gadolinium-enhanced MRI may be an indicator for surgery. PMID- 19565252 TI - Transforaminal lumbar interbody fusion with one cage and excised local bone. AB - INTRODUCTION: The effect of transforaminal lumbar interbody fusion (TLIF) with one cage and excised local bone were investigated in 52 patients with a mean follow-up of 18.2 months. METHOD: The clinical outcomes including the modified Prolo scale and a visual analog scale (VAS), and radiological assessments including the ratio of interbody graft area, fusion rate, posterior disk height (PH), and the lordosis angle (LA) of the motion segment were studied. RESULTS: According to a modified Prolo scale, 90.4% of the patients obtained either excellent or good results. The VAS significantly decreased postoperatively. There was significant postoperative improvement of the PH and LA, and no significant loss of the PH and LA was found at final follow-up. The fusion rate in this series was 96.6%. CONCLUSION: In conclusion, TLIF with one cage and excised local bone grafting can provide satisfactory treatment outcomes and solid interbody fusion without harvesting and grafting autologous iliac bone. PMID- 19565253 TI - Pelvic-peritoneal tuberculosis simulating peritoneal carcinomatosis: high clinical suspicion and a minimally invasive procedure. AB - Peritoneal tuberculosis (TB) is still a major health problem in developing countries. We describe a 26-year-old woman with peritoneal TB presenting with lower abdominal pain and distention, weight loss, and night sweats. There are no pathognomonic clinical, laboratory, or radiologic findings for peritoneal TB. Therefore, it can be easily confused with peritoneal carcinomatosis and advanced ovarian carcinoma. Adenosine deaminase activity in ascitic fluid combined with a high clinic suspicion is an effective and minimally invasive procedure for the differential diagnosis of pelvic-peritoneal TB simulating peritoneal carcinomatosis, and may obviate the need for unnecessary extensive surgery and rapidly initiate appropriate therapy. PMID- 19565254 TI - Severe endometriosis: laparoscopic rectum resection. AB - AIM: Endometriosis is a frequent benign disease of women in reproductive age. An infiltration of the spatium rectovaginal is rare, but if it occurs, in up to 73% the rectum is involved. If there is the indication for surgery, a partial resection of the rectum might be necessary. This can be performed by a laparoscopic approach. It is the aim of this work to describe a patient population treated for endometriosis in the spatium rectovaginal by laparoscopic surgery. PATIENT POPULATION AND METHODS: A retrospective analysis of data from patients with endometriosis in rectum or sigma, which underwent a laparoscopic partial bowel resection in the years 2005-2006 at the Department of Obstetrics and Gynecology, University of Regensburg, was carried out. RESULTS: Between 2005 and 2006, we performed a laparoscopic partial bowel resection in six patients with endometriosis. The mean age at diagnosis was 36.1 years (range 28-50 years) and 36.5 years (range 30-50 years) at surgery. All patients were nulligravida and 50% of the patients were infertile (since 1-6 years). The interval between the onset of symptoms and surgery ranged from a few weeks up to 2.5 years. Two-thirds of the patients had endocrine treatment before surgery. Three patients had a rectum resection, one a sigma resection and two had a combined rectum- and sigma resection. The mean duration of surgery was 201 min and mean hospital stay was 8 days. We saw one post-surgery bleeding at the enteroanastomosis. In that case two erythrocyte concentrates were necessary and the bleeding was stopped by rectoscopic intervention. All follow-up coloscopies were without pathological findings. One patient had a normal delivery after IVF/ICSI treatment. CONCLUSIONS: If severe endometriosis needs a rectum resection then it can be done laparoscopically. This surgery should be performed in a specialized center. The duration of surgery, hospital stay and time of convalescence are short. PMID- 19565255 TI - Pelvic actinomycosis confirmed after surgery: single center experience. AB - PURPOSE: To review pelvic actinomycosis confirmed by surgery and summarize clinical aspects. METHOD: The authors retrospectively reviewed 16 cases that underwent surgical procedures for pelvic actinomycosis between January 1998 and January 2006. RESULTS: Twelve of the cases (75%) had used intrauterine contraceptive devices for a period of between 8 months and 12 years (mean duration: 5.2 years). Most of the cases presented elevated white blood cell count, neutrophil percentage, CRP value and CA 125 value. Four of the eight cases that were checked by computed tomography presented the possibility of pelvic malignancies. All patients underwent surgery. The mean duration of intravenous antibiotics was 11.8 days and that of peroral antibiotics was 4.3 months. After treatment, there were no complications or recurrences, with the exception of one case of preoperative unilateral ureter obliteration. CONCLUSION: In diagnosis of pelvic actinomycosis, it is not necessary to consider whether an intrauterine device was being used or not. To distinguish from pelvic malignancy, various methods (including imaging study) should be considered. As for antibiotic treatment, it is believed that shorter and high dose antibiotics therapy will be sufficient treatment. PMID- 19565256 TI - Altered gene profile of placenta from women with intrahepatic cholestasis of pregnancy. AB - OBJECTIVES: To investigate the alterations in gene profile of placenta from pregnant women with intrahepatic cholestasis of pregnancy (ICP) and to enhance the insight of etiology and pathogenesis of ICP. METHODS: Ten pregnant women diagnosed ICP were recruited and 10 healthy pregnant women served as control. Four samples were taken from each placenta and RNA was isolated. Gene expression was analyzed with microarray and real time PCR was used to validate the differentially expressed genes. RESULTS: 392 genes were found differentially expressed. Among these differentially expressed genes, 280 were up-regulated and 112 were down-regulated. These differentially expressed genes involved 20 categories including genes involved in transportation, cell growth, apoptosis and immune response that were putatively participated the pathogenesis of ICP. CONCLUSIONS: 293 differentially expressed genes of 20 categories were found in ICP placenta, suggesting the diversity of gene expression alteration and the complexity of etiology and pathogenesis of ICP. PMID- 19565258 TI - Video-oculography findings and vestibular symptoms on the day of stapes surgery. AB - Vestibular symptoms are common after stapes surgery and may prevent outpatient treatment. The cause of vestibular symptoms is unclear, and in previous studies objective measurements have been obtained few days after the surgery. The aim of the study was to evaluate the existence and nature of any early, and possibly temporary vestibular irritation that occurs immediately after surgery. Twenty-one patients who had an opening of the oval window were prospectively included. Postoperative symptoms were collected and eye position curves were recorded with video-oculography (VOG) an average of 4 h after surgery. Early vestibular symptoms were found in 11 (52%) patients. Four patients had rotatory vertigo, three had floating sensation, one experienced a tilting sensation, and two had unspecific dizziness. Two patients vomited after surgery. When VOG was performed all but one were free of symptoms. After surgery, spontaneous nystagmus was found in three (14%) asymptomatic patients when gaze fixation was prevented. The prevalence of 33% (7 of 21 patients) of latent, spontaneous, horizontal-torsional nystagmus seen only during lateral gaze towards the direction of fast phase was significantly higher (P = 0.0001) than in healthy controls. The occurrence of nystagmus did not correlate with vestibular symptoms (P = 0.30). Slight, direction-fixed, latent, spontaneous horizontal-torsional nystagmus in some patients is consistent with a minor disturbance of semicircular canals, although it was not related to the sensation of dizziness. Recordings during stapes surgery may be needed to ascertain the origin of vestibular symptoms. Regarding vestibular symptoms and signs, patients may be safely discharged a few hours after stapes surgery. PMID- 19565259 TI - Inverted papilloma involving the frontal sinus: a management plan. AB - Inverted papilloma is a benign but locally aggressive sino-nasal tumour. Although relatively uncommon, involvement of the frontal sinus by this tumour represents a significant surgical challenge. The objective of the study is to propose a scheme for management of inverted papilloma involving the frontal sinus, based upon the findings of the current study. All cases of inverted papilloma operated upon between July 1995 and June 2008 were retrospectively reviewed to identify cases in which the tumour involved the frontal sinus. Among 34 patients with inverted papilloma, 4 were found to have tumours involving the frontal sinus (11.76%). These patients were initially treated by endonasal endoscopic resection. At time of initial surgical excision, the tumour was found to involve the frontal sinus by expansion from the ethmoids in three of these patients. In the fourth patient, the tumour was found to be massively involving the frontal sinus mucosa. After a mean follow-up of 16.3 months, no recurrences were detected in the first three patients. In the patient with massive mucosal involvement, recurrence was detected 4 years after the initial endonasal endoscopic resection. Subsequently, an osteoplastic flap was performed to resect the tumour. Fifty months later the patient remained disease free. Surgeons managing patients with frontal sinus inverted papilloma should have a clear management scheme before embarking on surgery. The patient's consent should be obtained pre-operatively for a possible osteoplastic flap. Tumours just expanding into the frontal sinus can be managed by either endoscopic or nonendoscopic approaches. On the other hand, in tumours significantly involving the frontal sinus mucosa, an osteoplastic flap is warranted to ensure complete tumour resection. PMID- 19565260 TI - Brachial artery retrograde flow increases with age: relationship to physical function. AB - The purpose of this study was to examine the flow velocity pattern of the brachial artery and to determine its relationship to measures of physical function. Subjects from the Louisiana Healthy Aging Study (n = 95; age = 84 +/- 10 years) were evaluated. Brachial artery flow velocities and dimensions were measured using high-resolution ultrasonography. The continuous scale of physical function and performance test (CS-PFP10) was used to assess physical function. This test is based on the performance of 11 activities of daily living. Total CS PFP10 score was 39.51 +/- 21.21 U. Mean antegrade and retrograde velocities at rest were 14.2 +/- 4.7 and 3.6 +/- 2.2 cm/s, respectively. Ante-/retrograde ratio was 5.5 +/- 4.6. Brachial artery diameter was 4.3 +/- 0.7 mm. Pulse pressure and vascular conductance were 66 +/- 18 mmHg, and 0.9 +/- 0.5 ml/min/mmHg, respectively. Vascular conductance (r = -0.34), ante-/retrograde ratio (r = 0.42) and CS-PFP10 (r = -0.65) were inversely and retrograde velocity (r = 0.40) and pulse pressure (r = 0.36), were directly associated with age. Retrograde velocity was inversely related to vascular conductance (r = -0.27) and CS-PFP10 total score (r = -0.45). A MANOVA revealed that those with the higher CS-PFP10 scores had a lower retrograde velocity (P = 0.0001), but this association was, in part, age-dependent. Among nonagenarians (n = 52), those in the lower tertiles of the CS-PFP10 scores had significantly higher retrograde velocities compared to those in the higher tertiles (P = 0.035). These data indicate an increase in brachial retrograde velocity with age. These hemodynamic changes are related to a decline in physical function. PMID- 19565261 TI - Phylogenetic and evolutionary relationships and developmental expression patterns of the zebrafish twist gene family. AB - Four members of the twist gene family (twist1a, 1b, 2, and 3) are found in the zebrafish, and they are thought to have arisen through three rounds of gene duplication, two of which occurred prior to the tetrapod-fish split. Phylogenetic analysis groups most of the vertebrate Twist1 peptides into clade I, except for the Twist1b proteins of the acanthopterygian fish (medaka, pufferfish, stickleback), which clustered within clade III. Paralogies and orthologies among the zebrafish, medaka, and human twist genes were determined using comparative synteny analysis of the chromosomal regions flanking these genes. Comparative nucleotide substitution analyses also revealed a faster rate of nucleotide mutation/substitution in the acanthopterygian twist1b compared to the zebrafish twist1b, thus accounting for their anomalous phylogenetic clustering. We also observed minimal expression overlap among the four twist genes, suggesting that despite their significant peptide similarity, their regulatory controls have diverged considerably, with minimal functional redundancy between them. PMID- 19565262 TI - Greater than the sum of its parts: a review of studies combining structural connectivity and resting-state functional connectivity. AB - It is commonly assumed that functional brain connectivity reflects structural brain connectivity. The exact relationship between structure and function, however, might not be straightforward. In this review we aim to examine how our understanding of the relationship between structure and function in the 'resting' brain has advanced over the last several years. We discuss eight articles that directly compare resting-state functional connectivity with structural connectivity and three clinical case studies of patients with limited white matter connections between the cerebral hemispheres. All studies examined show largely convergent results: the strength of resting-state functional connectivity is positively correlated with structural connectivity strength. However, functional connectivity is also observed between regions where there is little or no structural connectivity, which most likely indicates functional correlations mediated by indirect structural connections (i.e. via a third region). As the methodologies for measuring structural and functional connectivity continue to improve and their complementary strengths are applied in parallel, we can expect important advances in our diagnostic and prognostic capacities in diseases like Alzheimer's, multiple sclerosis, and stroke. PMID- 19565263 TI - An uncommon cause of abdominal pain and diarrhea-gastrinoma in an adolescent. AB - Gastrinoma is a hormone-secreting tumor associated with the Zollinger-Ellison syndrome. It is quite rare among children, and it is also uncommon in locations other than the pancreas and the duodenum in the pediatric group. Here, we describe an adolescent male, presenting with recurrent secretory diarrhea and abdominal cramping pain, who had a solitary gastrinoma in the lesser sac, close to the stomach. The prognosis was good after complete resection. PMID- 19565264 TI - The prognostic role of phospho-Src family kinase analysis in tongue cancer. AB - BACKGROUND: The up regulation of Phospho-Src family kinase oncogene has been correlated with reduced postoperative survival in various cancers but never in tongue cancer. METHODS: We analyzed phospho-Src family kinase in 39 tongue (mobile) cancer patients by immunohistochemistry, compared these results with similar analysis for TUNEL and c-erbB-2 and with both clinical tumor characteristics and patient survival probability rates. RESULTS: Phospho-Src family kinase overexpression was found in most tongue cancer biopsies (62%), significantly correlating with tumors larger in size (P = 0.05), progression lymph node metastasis (0.004) and stage (P = 0.05), and correlating with TUNEL (P = 0.01) and c-erbB-2 (P = 0.05) expression rates. At 60 months, survival probability for negative phospho-Src family kinase level (=0) patients was 67%, but 30% for positive phospho-Src family kinase level (>0) patients (P = 0.05). CONCLUSIONS: Inverse correlation between phospho-Src family kinase and patient survival demonstrates the prognostic role of phospho-Src family kinase in tongue cancer. These findings suggest a novel link between phospho-Src family kinase and TUNEL and c-erbB-2 pathways, tilting the balance toward cell proliferation. PMID- 19565266 TI - Hypermethylated SFRP1, but none of other nine genes "informative" for western countries, is valuable for bladder cancer detection in Mainland China. AB - PURPOSE: A 11-gene set by methylation-specific PCR in urine sediments for sensitive/specific detection of bladder cancer has been identified previously. In this study, we have evaluated 10 DNA methylation biomarkers that have been reported informative in western countries for bladder cancer diagnosis for a better set. MATERIALS AND METHODS: The promoter CpG Islands of the following 10 genes: CDH1, FANCF, LOXL1, LOXL4, p16INK4, SFRP1, SOX9, TIG1, TIMP3, and XAF1 have been subjected to methylation-specific PCR analysis in the DNA of 2 bladder cancer cell lines, 2 normal bladder tissues and urine sediments of 82 bladder cancer patients, 15 non-cancerous urogenital patients and 5 healthy volunteers. RESULTS: Both XAF1 and LOXL1 genes were heterozygously methylated in the normal bladder tissues, showing no cancer state specificity. While the hypermethylated states were detected in urine sediments of bladder cancer at a frequency not less than 2.4% (2/82 cases), nine genes were also methylated in the patients of the non-cancerous urogenital diseases. The methylated SFRP1 was detected in 36.6% (30/82 cases) of bladder cancer and 6.7% (1/15 cases) of non-cancerous urogenital diseases, showing the bladder cancer specificity. CONCLUSIONS: Inclusion of the SFRP1 gene into a set of 11 genes has improved the bladder cancer detection. The insufficiency of predicting disease onset in this study with the previously recommended targets in western countries suggests a possible disease disparity between these two populations. Alternatively, the tissue-specific methylation might be mistaken as the cancer specific in the studies where no non-cancerous lesion controls were involved. PMID- 19565267 TI - Molecular survey of pyrethroid resistance mechanisms in Mexican field populations of Rhipicephalus (Boophilus) microplus. AB - Susceptibility to synthetic pyrethroids (SP s) and the role of two major resistance mechanisms were evaluated in Mexican Rhipicephalus microplus tick populations. Larval packet test (LPT), knock-down (kdr) PCR allele-specific assay (PASA) and esterase activity assays were conducted in tick populations for cypermethrin, flumethrin and deltamethrin. Esterase activity did not have a significant correlation with SP s resistance. However a significant correlation (p < 0.01) was found between the presence of the sodium channel mutation, and resistance to SP s as measured by PASA and LPT respectively. Just over half the populations (16/28) were cross-resistant to flumethrin, deltamethrin and cypermethrine, 21.4% of the samples (6/28) were susceptible to all of the three pyrethroids 10.7 of the samples (3/28) were resistant to flumethrin, 3.4 of the samples (1/28) were resistant to deltamethrin only and 7.1% (2/28) were resistant to flumethrin and deltamethrin. The presence of the kdr mutation correlates with resistance to the SP s as a class. Target site insensitivity is the major mechanism of resistance to SP s in Mexican R. microplus field strains, involving the presence of a sodium channel mutation, however, esterase-based, other mutations or combination of mechanisms can also occur. PMID- 19565269 TI - Preliminary studies on the isolation of bacteria from sea lice, Lepeophtheirus salmonis, infecting farmed salmon in British Columbia, Canada. AB - Using standard OIE bacteriological screening protocols, we sampled the external carapace and internal stomach contents of motile stages (preadult and adult) of Lepeophtheirus salmonis collected from farmed Atlantic salmon from May 2007 to April 2008 in British Columbia, Canada. Three potentially pathogenic bacteria (Tenacibaculum maritimum, Pseudomonas fluorescens, and Vibrio spp.) were isolated from external (58-100%) and internal (12.5-100%) samples of sea lice. The prevalence of bacteria was higher from lice collected during the months with higher water temperatures and among adult lice. These preliminary results have led to a comprehensive, multi-year study where we plan to examine the possible role of sea lice as a vector for disease. PMID- 19565268 TI - Isolation and genotyping of potentially pathogenic Acanthamoeba and Naegleria species from tap-water sources in Osaka, Japan. AB - Here, we carried out a survey to determine the prevalence of free-living amoebae (FLA) in tap-water sources from rivers and water treatment plants located in Osaka Prefecture, Japan. A total of 374 raw water samples were collected from 113 sampling points. The samples were filtrated and transferred to non-nutrient agar plates seeded with a heat-killed suspension of Escherichia coli and incubated for 2 to 7 days at 30 degrees C or 42 degrees C. The plates were examined by microscopy to morphologically identify FLA families, and polymerase chain reaction and sequence analysis were then performed to define the species of the detected Naegleria and Acanthamoeba isolates. A total of 257 of 374 samples (68.7%) were positive for FLA by microscopy, and among these there were 800 FLA isolates, including Acanthamoeba and Naegleria species. Sequence analysis identified five Acanthamoeba spp. isolates of the known pathogenic T4 genotype and 43 Naegleria australiensis isolates, a reported pathogen to mice and also of concern as a potential pathogen to humans. Our results suggest a wide distribution of FLA, including potential pathogenic species, in tap-water sources of western Japan. PMID- 19565270 TI - EST sequencing of blood-fed and Leishmania-infected midgut of Lutzomyia longipalpis, the principal visceral leishmaniasis vector in the Americas. AB - Leishmaniasis is an important worldwide public health problem. Visceral leishmaniasis caused by Leishmania infantum chagasi is mainly transmitted by Lutzomyia longipalpis in the Americas. Leishmania development within the sand fly vector is mostly restricted to the midgut. Thus, a comparative analysis of blood fed versus infected midguts may provide an invaluable insight into various aspects of sand fly immunity, physiology of blood digestion, and, more importantly, of Leishmania development. To that end, we have engaged in a study to identify expressed sequenced tags (ESTs) from L. longipalpis cDNA libraries produced from midguts dissected at different times post blood meal and also after artificial infection with L. i. chagasi. A total of 2,520 ESTs were obtained and, according to the quality of the sequencing data obtained, assembled into 378 clusters and 1,526 individual sequences or singletons totalizing 1,904 sequences. Several sequences associated with defense, apoptosis, RNAi, and digestion processes were annotated. The data presented here increases current knowledge on the New World sand fly transcriptome, contributing to the understanding of various aspects of the molecular physiology of L. longipalpis, and mechanisms underlying the relationship of this sand fly species with L. i. chagasi. PMID- 19565273 TI - Ho Nam Chang special issue: life of a great biochemical engineer and his life time contribution to high cell density culture. PMID- 19565271 TI - The cannabinoid receptor type 2 (CNR2) gene is associated with hand bone strength phenotypes in an ethnically homogeneous family sample. AB - Genetic variants within the CNR2 gene encoding the cannabinoid receptor CB2 have been shown to be associated with osteoporosis and low bone mineral density (BMD) in case-control studies. We now examined the association of polymorphisms in CNR2 with hand bone strength in an ethnically homogeneous healthy family sample of European origin (Chuvashians) living in Russia. We show that non-synonymous CNR2 SNPs are significantly associated with radiographic hand BMD and breaking bending resistance index (BBRI) by two different transmission disequilibrium tests. For both tests highly significant p values (ranging from 0.007 to 0.008 for hand BMD, and from 0.001 to 0.003 for BBRI) were also obtained with additional SNPs at the CNR2 locus. The associations remained significant after correction for multiple testing. In conclusion, in addition to the association of CNR2 polymorphisms with low BMD at selected clinically relevant skeletal sites, we now report their significant association with hand bone strength phenotypes using a family-based study design implying an even broader impact of genetic variation at the CNR2 locus on bone structure and function. PMID- 19565274 TI - An infant presenting with a non-functional kidney on dimercaptosuccinic acid scan: answer. PMID- 19565275 TI - An infant presenting with a non-functional kidney on dimercaptosuccinic acid scan: question. PMID- 19565276 TI - A qualitative study of decision-making by breast cancer patients about telling their children about their illness. AB - OBJECTIVE: Many breast cancer patients are troubled about telling their school age children about their illness. However, little attention has been paid to the factors that encourage or discourage them from revealing the illness. This study explored decision-making by breast cancer patients about telling their children about their illness. METHODS: Participants were 30 breast cancer patients recruited from a regional cancer institution in Japan. Semi-structured interviews were conducted and content analysis was performed. RESULTS: Six preparatory stages of decision-making by Japanese breast cancer patients about telling their children about their illness were identified as follows: contemplation, preparation, action-hospitalization and surgery, action-adjuvant therapy, action diagnosis, and action-prognosis. We also identified 11 categories of positive aspects and ten categories of negative aspects about revealing their illness to children. The categories of negative aspects with higher frequency were similar to those found by previous research, but categories of positive aspects were unique. The rate of reference to negative aspects in total reduces gradually as the preparatory stage advances, and in action-diagnosis and action-prognosis stages the balance between positive and negative aspects becomes about half and half. CONCLUSIONS: Patients, especially in action-hospitalization and surgery, can be expected to tell their children about their illness although they find negative aspects much more compelling than positive aspects and experience great distress. These patients have special needs for support from others. PMID- 19565277 TI - Comparison of risk factors predicting return to work between patients with subacute and chronic non-specific low back pain: systematic review. AB - The objective of the study was to provide an inventory of predictive instruments and their constituting parameters associated with return to work in patients with subacute (2-10 weeks pain duration) and chronic (10-24 weeks pain duration) non specific low back pain (NSLBP). Data sources included systematic review in Medline, Embase, Cinahl, Central, PEDro, Psyndex, PsychInfo/PsycLit, and Sociofile up to September 2008, in reference lists of systematic reviews on risk factors, and of included studies. For the systematic review, two reviewers independently assessed study eligibility and quality, and extracted data. Disagreements were resolved by consensus. Risk factors were inventorised and grouped into a somatic and psychosocial domain. 23 studies reporting on subacute and 16 studies reporting on chronic patients were included. The studies on subacute patients reported on a total of 56 biomedical factors out of which 35 (63%) were modifiable and 61 psychosocial factors out of which 51 (84%) were modifiable. The corresponding values in studies on chronic patients were 44 biomedical [27 (62%) modifiable] and 61 [40 (66%) modifiable] respectively. Our data suggest that the interdisciplinary approach in patients at risk to develop persistent NSLBP is justified in both, the subacute and chronic disease stages. Psychosocial interventions might be more effective in subacute stages since a higher proportion of modifiable risk factors were identified in that group. PMID- 19565278 TI - Comparative biomechanical investigation of a modular dynamic lumbar stabilization system and the Dynesys system. AB - The goal of non-fusion stabilization is to reduce the mobility of the spine segment to less than that of the intact spine specimen, while retaining some residual motion. Several in vitro studies have been conducted on a dynamic system currently available for clinical use (Dynesys). Under pure moment loading, a dependency of the biomechanical performance on spacer length has been demonstrated; this variability in implant properties is removed with a modular concept incorporating a discrete flexible element. An in vitro study was performed to compare the kinematic and stabilizing properties of a modular dynamic lumbar stabilization system with those of Dynesys, under the influence of an axial preload. Six human cadaver spine specimens (L1-S1) were tested in a spine loading apparatus. Flexibility measurements were performed by applying pure bending moments of 8 Nm, about each of the three principal anatomical axes, with a simultaneously applied axial preload of 400 N. Specimens were tested intact, and following creation of a defect at L3-L4, with the Dynesys implant, with the modular implant and, after removal of the hardware, the injury state. Segmental range of motion (ROM) was reduced for flexion-extension and lateral bending with both implants. Motion in flexion was reduced to less than 20% of the intact level, in extension to approximately 40% and in lateral bending a motion reduction to less than 40% was measured. In torsion, the total ROM was not significantly different from that of the intact level. The expectations for a flexible posterior stabilizing implant are not fulfilled. The assumption that a device which is particularly compliant in bending allows substantial intersegmental motion cannot be fully supported when one considers that such devices are placed at a location far removed from the natural rotation center of the intervertebral joint. PMID- 19565279 TI - Oral biofilm models for mechanical plaque removal. AB - In vitro plaque removal studies require biofilm models that resemble in vivo dental plaque. Here, we compare contact and non-contact removal of single and dual-species biofilms as well as of biofilms grown from human whole saliva in vitro using different biofilm models. Bacteria were adhered to a salivary pellicle for 2 h or grown after adhesion for 16 h, after which, their removal was evaluated. In a contact mode, no differences were observed between the manual, rotating, or sonic brushing; and removal was on average 39%, 84%, and 95% for Streptococcus mutans, Streptococcus oralis, and Actinomyces naeslundii, respectively, and 90% and 54% for the dual- and multi-species biofilms, respectively. However, in a non-contact mode, rotating and sonic brushes still removed considerable numbers of bacteria (24-40%), while the manual brush as a control (5-11%) did not. Single A. naeslundii and dual-species (A. naeslundii and S. oralis) biofilms were more difficult to remove after 16 h growth than after 2 h adhesion (on average, 62% and 93% for 16- and 2-h-old biofilms, respectively), while in contrast, biofilms grown from whole saliva were easier to remove (97% after 16 h and 54% after 2 h of growth). Considering the strong adhesion of dual species biofilms and their easier more reproducible growth compared with biofilms grown from whole saliva, dual-species biofilms of A. naeslundii and S. oralis are suggested to be preferred for use in mechanical plaque removal studies in vitro. PMID- 19565281 TI - How great is great ape foresight? AB - Osvath and Osvath (Anim Cogn 11: 661-674, 2008) report innovative studies with two chimpanzees and one orangutan that suggest some capacity to select and keep a tool for use about an hour later. This is a welcome contribution to a small, but rapidly growing, field. Here we point out some of the weaknesses in the current data and caution the interpretation the authors advance. It is not clear to what extent the apes really engaged in any foresight in these studies. PMID- 19565280 TI - A modifier locus on chromosome 5 contributes to L1 cell adhesion molecule X linked hydrocephalus in mice. AB - Humans with L1 cell adhesion molecule (L1CAM) mutations exhibit X-linked hydrocephalus, as well as other severe neurological disorders. L1-6D mutant mice, which are homozygous for a deletion that removes the sixth immunoglobulin-like domain of L1cam, seldom display hydrocephalus on the 129/Sv background. However, the same L1-6D mutation produces severe hydrocephalus on the C57BL/6J background. To begin to understand how L1cam deficiencies result in hydrocephalus and to identify modifier loci that contribute to X-linked hydrocephalus by genetically interacting with L1cam, we conducted a genome-wide scan on F2 L1-6D mice, bred from L1-6D 129S2/SvPasCrlf and C57BL/6J mice. Linkage studies, utilizing chi square tests and quantitative trait loci mapping techniques, were performed. Candidate modifier loci were further investigated in an extension study. Linkage was confirmed for a locus on chromosome 5, which we named L1cam hydrocephalus modifier 1 (L1hydro1), p = 4.04 X 10(-11). PMID- 19565283 TI - On the dimensionality of the System Usability Scale: a test of alternative measurement models. AB - The System Usability Scale (SUS), developed by Brooke (Usability evaluation in industry, Taylor & Francis, London, pp 189-194, 1996), had a great success among usability practitioners since it is a quick and easy to use measure for collecting users' usability evaluation of a system. Recently, Lewis and Sauro (Proceedings of the human computer interaction international conference (HCII 2009), San Diego CA, USA, 2009) have proposed a two-factor structure-Usability (8 items) and Learnability (2 items)-suggesting that practitioners might take advantage of these new factors to extract additional information from SUS data. In order to verify the dimensionality in the SUS' two-component structure, we estimated the parameters and tested with a structural equation model the SUS structure on a sample of 196 university users. Our data indicated that both the unidimensional model and the two-factor model with uncorrelated factors proposed by Lewis and Sauro (Proceedings of the human computer interaction international conference (HCII 2009), San Diego CA, USA, 2009) had a not satisfactory fit to the data. We thus released the hypothesis that Usability and Learnability are independent components of SUS ratings and tested a less restrictive model with correlated factors. This model not only yielded a good fit to the data, but it was also significantly more appropriate to represent the structure of SUS ratings. PMID- 19565282 TI - Production of L-phenylalanine from glycerol by a recombinant Escherichia coli. AB - The production of L-phenylalanine is conventionally carried out by fermentations that use glucose or sucrose as the carbon source. This work reports on the use of glycerol as an inexpensive and abundant sole carbon source for producing L phenylalanine using the genetically modified bacterium Escherichia coli BL21(DE3). Fermentations were carried out at 37 degrees C, pH 7.4, using a defined medium in a stirred tank bioreactor at various intensities of impeller agitation speeds (300-500 rpm corresponding to 0.97-1.62 m s(-1) impeller tip speed) and aeration rates (2-8 L min(-1), or 1-4 vvm). This highly aerobic fermentation required a good supply of oxygen, but intense agitation (impeller tip speed approximately 1.62 m s(-1)) reduced the biomass and L-phenylalanine productivity, possibly because of shear sensitivity of the recombinant bacterium. Production of L-phenylalanine was apparently strongly associated with growth. Under the best operating conditions (1.30 m s(-1) impeller tip speed, 4 vvm aeration rate), the yield of L-phenylalanine on glycerol was 0.58 g g(-1), or more than twice the best yield attainable on sucrose (0.25 g g(-1)). In the best case, the peak concentration of L-phenylalanine was 5.6 g L(-1), or comparable to values attained in batch fermentations that use glucose or sucrose. The use of glycerol for the commercial production of L-phenylalanine with E. coli BL21(DE3) has the potential to substantially reduce the cost of production compared to sucrose- and glucose-based fermentations. PMID- 19565284 TI - Chemosensitivity and survival in gastric cancer patients with microsatellite instability. AB - INTRODUCTION: Conflicting data exist regarding the relevance of high-frequency microsatellite instability (MSI-H) for predicting the prognosis and benefits of 5 fluorouracil (5-FU)-based chemotherapy. This study investigated the usefulness of MSI as either a prognostic indicator or predictor of distinct clinical attributes regarding the use of adjuvant chemotherapy with 5-FU and its analogues in gastric cancer. MATERIALS AND METHODS: Data and tumor specimens were collected from 240 gastric cancer patients from 1993 to 2002. Five microsatellite loci were analyzed using a high-intensity microsatellite analysis reported previously. A Cox proportional hazard model was used to compare the clinical data and survival as well as any associations between MSI and 5-FU treatment status of patients with MSI or microsatellite stability (MSS) gastric cancers. A 3-(4,5-dimethyl-2 thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was conducted in 168 cases to investigate chemosensitivity to 5-FU. RESULTS: This analysis identified 22 MSI-H (9.4%), 25 MSI-L (10.7%), and 193 MSS (79.9%) tumors. Gastric cancer with MSI-H tended to have increased likelihood to show higher age, antral location of the tumor, and lymph vessel involvement (P < 0.05). Univariate analyses failed to show any difference between the MSI-H and MSS/MSI-L groups with respect to overall survival. Furthermore, survival after the administration of 5-FU did not correlate with MSI status, and MSI was not associated with 5-FU sensitivity by MTT assay. CONCLUSION: The results of this study indicate that MSI status has no clear influence on overall survival or response to 5-FU in gastric cancer. PMID- 19565286 TI - High rate of multifocality and occult lymph node metastases in papillary thyroid carcinoma arising in thyroglossal duct cysts. AB - BACKGROUND: The incidence of cancer in thyroglossal duct cysts (TDC) is low and management is controversial. The objective was to report the rate of multifocality, lymph node metastases, and long-term results for TDC carcinomas in adults. MATERIALS AND METHODS: Files from 1979 to 2008 were reviewed for tumor stage, multifocality in the lobes, lymph node metastases, treatment, and follow up. RESULTS: A total of 18 patients (13 females, 5 males, average age 41.5 years) were treated for papillary carcinoma arising in a TDC. Of these, 15 underwent total thyroidectomy, 1 isthmusectomy and 2 a Sistrunk procedure only. Also, 16 patients underwent neck dissection of the central and/or lateral compartments. Tumors were staged pT1 (n = 15), pT3 (n = 3), pN0 (n = 4), pN1a (n = 3), pN1b (n = 9), Nx (n = 2), M0 (n = 17), and M1 (n = 1, lung metastases). Tumor foci were found in the thyroid lobes in 9 of 16 patients(56%). Lymph node metastases were found in 12 of 16 (75%). Nodes were positive in 6 of 15 central compartment dissections (40%) and in 9 of 15 lateral neck dissections (60%). Metastases to the lateral compartment, with no central compartment metastasis, were found in 6 of 15 patients (40%). Radioiodine was administered to 12 patients. Median follow up was 12 years (range 1-22 years). All had negative ultrasound. Stimulated Tg levels available for 11 patients were undetectable for 10 and 2 ng/mL for the remaining patient. CONCLUSIONS: This series shows a high rate of thyroid lobe foci and lymph node metastases but an excellent long-term outcome, characteristics shared with classic papillary carcinoma. Lateral neck metastases seem to be more frequent. These findings are in favor of following the current guidelines for differentiated thyroid cancer in general for the treatment of these rare tumors. PMID- 19565287 TI - One-staged subtotal sacrectomy for primary sacral tumor. AB - BACKGROUND: Sacrectomy with adequate margins is challenging because of the complexity of the surgical approach and morbidities. Two-staged sequential approach, therefore, has been advocated. This study was designed to demonstrate the modification of this technique. METHODS: This is a case presentation of a 45 year-old man with chordoma involving the lower border of S2, who underwent one staged subtotal sacrectomy. The technique involved the following: midline incision, mobilization of the rectum, construction of a colostomy and the modified Hartmann stump with intact superior rectal vessels, ligation of internal iliac arteries, ligation of all branches connecting to external iliac veins resulting in "complete isolation" of the external iliac veins, dissection of presacral tissue, anterior osteotomy at the S1-S2 junction and the sacroiliac joints, and abdominal closure. The posterior approach involved a three-limbed incision, dissection of the gluteus muscle and ligaments from the sacrum, subperiosteal dissection, S1 laminectomy, posterior osteotomy corresponding with the anterior osteotomy line with preservation of S1 nerves, division of S2-S4 nerves from sciatic nerves, and specimen removal. Closure of the large sacral defect was undertaken using the Hartmann stump and bilateral gluteus maximus flaps. RESULTS: En bloc resection with free margins without tumor rupture was accomplished. Operative time was 12 hours. Blood transfusion was 6 units. This patient had a good recovery without complications. He was able to ambulate within 1 week and walk normally within 1 month. No recurrence was found at a 24-month follow-up. CONCLUSIONS: One-staged sacrectomy can be safely performed, obtaining the satisfactory outcomes. PMID- 19565285 TI - Serum p53 antibody as tumor marker for follow-up of colorectal cancer after curative resection. AB - BACKGROUND: No large-scale studies have examined the use of serial measurements of serum p53 antibodies (s-p53Abs) combined with carcinoembryonic antigen (CEA) measurements during the follow-up of colorectal cancer (CRC) patients after curative resection. METHODS: A highly specific enzyme-linked immunosorbent assay was used to analyze s-p53Abs levels in 305 CRC patients before and after curative resection at a single institution. Agreement between recurrence and serial s p53Ab and CEA measurements was evaluated by diagnostic accuracy odds ratio (DOR), kappa, and area under receiver operating characteristic curve (AUC). RESULTS: Among 305 patients, 76 (25%) patients had disease recurrence during follow-up. None of the 168 s-p53Ab seronegative patients (s-p53Ab < 10 U/microL) without recurrence had an abnormal s-p53Ab test during follow-up. Among the remaining low level (10 U/microL 76 U/microL, n = 34) seropositive patients, recurrence defined by s p53Ab tests resulted in a DOR of 4.3 and infinity, a kappa of 0.35 and 1.00, and an AUC of 0.633 [95% confidence interval (CI), 0.495 to 0.772; P = 0.047], and 1.0 (95% CI, 1.000 to 1.000; P < 0.0001), respectively. Recurrence defined by CEA tests had an AUC of 0.781 (95% CI, 0.654 to 0.909) for low-level and 0.796 (95% CI, 0.611 to 0.982) for high-level seropositive patients. CONCLUSIONS: Agreement between clinical recurrence and serial s-p53Ab test was dependent upon preoperative s-p53Ab level. Serial s-p53Ab testing outperformed CEA testing when predicting clinical recurrence in colorectal cancer patients with an abnormal preoperative s-p53Ab level. PMID- 19565290 TI - Sleep technologist performance: a call for standardization and performance feedback. PMID- 19565291 TI - Standardization of quality assurance for sleep technologist: a model. AB - INTRODUCTION: Since the last decade, there has been a tremendous growth of sleep centers in the US to meet the increasing need of diagnosing and treating sleep disorders. However, this unregulated growth has resulted in tremendous variance in the quality of sleep centers across the nation. The American Academy of Sleep Medicine, in an attempt to provide a benchmark standard, has introduced a voluntary accreditation process, part of which involves assessment of technical quality parameters. However, measuring technical quality is not easy. HYPOTHESIS: We undertook a study to determine if the implementation of point system and schematic feedback on technologist performance can result in improvement and tracking of their performance. MATERIALS AND METHODS: We randomly reviewed 100 charts from the preimplementation phase as control and 1,739 charts from the post implementation of the point system phase as study group. RESULTS: There was a statistically significant difference in the score among technologist between the control and study groups with the average being 75 +/- 4.12 and 87.53 +/- 0.91, respectively, with a p value being 0.0001. CONCLUSION: Evaluating the performance of the sleep technologist can be a way to track and monitor their performance in a standardized way and to identify weakness at an earlier stage. We present a system, which we have developed and implemented at our sleep center, as a possible model of assessing and subsequently standardizing technical quality for polysomnography. PMID- 19565292 TI - [Neuro-enhancement. Brain doping]. AB - Cognitive enhancement, the increase in the mental ability by psychoactive substances and other interventions has received a renewed boost through the development of innovative principle. More than 100 drugs are currently being developed, tested or used for cognitive enhancement. Cholinesterase inhibitors, memantine, dimebon, ampakines, fluoxetine and other antidepressants, methylphenidate and modafinil are candidates awaiting a larger distribution as cognitive enhancers in healthy individuals, if their advantages can be demonstrated. Beyond more general neuro-ethical reservations regarding neuro enhancement, future research will need to address the following neuropsychiatric issues: (1) will the benefits of longer term neuro-enhancement outweigh potential disadvantages such as rebound effects and other neurobiological and psychosocial trade-offs? (2) What will be the neuropsychiatric sequelae of a soft coercion towards drug usage at work and for recreational purposes? (3) Will there be new and specific neuropsychiatric diseases due to long-term usage of neuro-enhancers in a larger population? Novel strategies of neuro-enhancement will have to demonstrate their superiority compared with more traditional and well-established interventions such as coffee and cake. PMID- 19565295 TI - Endoscopic closure of gastric access in perspective NOTES: an update on techniques and technologies. AB - BACKGROUND: Natural orifice transluminal endoscopic surgery (NOTES) is currently of major research interest as it may offer significant clinical potential for endoscopic procedures in the future. Nevertheless, many issues are still unresolved. The aim of this study is a systematic review of the literature to evaluate techniques and technologies to address the issue of achieving a secure gastric wall defect closure. METHODS: Studies were identified by searching MEDLINE, EMBASE, Current Contents, Cochrane Library, Entrez PubMed and Clinical Trials Database until December 2008. We divided the different technologies into four groups: clipping, stitching, stapling and occluding systems. We analysed the different technologies in terms of (1) simplicity of manoeuvring for port closure, (2) security of introduction, (3) permanent implantation of the device into the organ, (4) effectiveness of port closure and (5) availability on the market. RESULTS: Twenty-eight published studies were identified, while nine clinical trials are ongoing. Among proposed technologies, staplers and occluders have greater effectiveness but lack of security, and in the case of staplers also lack of simplicity of use. None of the stitching technologies can really be considered effective except for the Flexible Endostitch, which on the other side is difficult to handle in the current fashion. Standard clips, although easy to handle, have never been proved to be reliable, as only mucosal layer closure has been demonstrated to date. On the other side, the over-the-scope clip is likely to be effective, secure to introduce and simple to handle within the hollow organ. CONCLUSIONS: Achieving reliable closure of the hollow organ defect represents the first step towards implementation of clinical NOTES. The current data do not yet allow definitive and clear advantages or disadvantages of the different options to be determined. The introduction of the EURONOTES registry will help us to understand in which direction to proceed. PMID- 19565296 TI - Combined percutaneous-endoscopic stenting of malignant biliary obstruction: results from 106 consecutive procedures and identification of factors associated with adverse outcome. AB - BACKGROUND: In patients in whom attempted endoscopic stenting of malignant biliary obstruction fails, combined percutaneous-endoscopic stenting and percutaneous stenting using expandable metallic endoprostheses offer alternative approaches to biliary drainage. Despite the popularity of the percutaneous route, there is no available evidence to support its superiority over combined stenting in this patient group. The objective of this study was to present the short- and long-term results of a large series of combined percutaneous-endoscopic stenting procedures and identify factors associated with adverse outcome. METHODS: Data were retrospectively collected on patients undergoing combined percutaneous endoscopic biliary stenting for malignant biliary obstruction between January 2002 and December 2006. Short- and long-term outcomes were recorded, and pre procedure variables correlated with adverse outcome. RESULTS: Combined biliary stenting was technically successful in 102 (96.2%) of 106 patients. Procedure associated mortality rate was 0%. In-hospital morbidity and mortality rates were 24.5% and 16.7%, respectively, with the majority of deaths resulting from biliary sepsis. Median survival was 100 days, with a 13.7% stent occlusion rate. On multivariable analysis, baseline American Society of Anaesthesiologists (ASA) grade, decreasing serum albumin and increasing leucocyte count were independently associated with in-hospital mortality following combined stenting. CONCLUSION: Combined biliary stenting is associated with short- and long-term outcomes equal to those reported in recent series of percutaneous transhepatic stenting. Randomised control trials, including cost-effectiveness analyses, are required to further compare these techniques. Outcomes following combined stenting may be further improved by early recognition and treatment of sepsis and scrupulous management of co-morbid disease. PMID- 19565297 TI - Treatment of rectal adenomas by transanal endoscopic microsurgery: 15 years' experience. AB - BACKGROUND: The authors present their experience with rectal adenomas managed by transanal endoscopic microsurgery (TEM). The goals of this study were to examine our institution's experience by evaluating surgical morbidity, mortality, and local recurrence rate. METHODS: This retrospective study investigated 402 patients who underwent TEM a for preoperative diagnosis of adenoma from January 1993 to October 2008. The mean age was 65 years (range = 22-92 years). All patients were regularly followed up to determine treatment efficacy in terms of local recurrence rate. RESULTS: No 30-day perioperative mortality occurred. No conversion to laparoscopic or open procedures was reported. Minor complications occurred in 28 (7%) patients, whereas major complications were found only in 2 (0.5%) patients. Definitive histology confirmed adenomas in 366 cases (91%). At a mean follow-up of 84 months (range = 1-190 months), 16 (4%) adenomas recurred and were successfully retreated by TEM [14 cases (87.5%)] and by conventional surgery [2 patients (12.5%)]. No further recurrences were observed at subsequent follow up. CONCLUSION: The findings warrant the conclusion that TEM is a safe, effective treatment for rectal adenomas where endoscopic removal is not applicable and has low morbidity and no mortality. PMID- 19565298 TI - Partial amniotic carbon dioxide insufflation (PACI) during minimally invasive fetoscopic surgery: early clinical experience in humans. AB - BACKGROUND: The technical performance of minimally invasive fetoscopic surgery may be severely hindered by poor visualization of intra-amniotic contents. Partial amniotic carbon dioxide insufflation (PACI) allows the visual limitations of operating within the fluid environment to be overcome. PATIENTS AND METHODS: When amniotic fluid exchange failed to improve fetoscopic visualization, PACI was attempted during 37 fetoscopic procedures between 17 + 5 and 33 + 2 weeks of gestation. PACI was attempted with filtered carbon dioxide using a commercially available insufflator via one to three trocars that were percutaneously introduced into the amniotic cavity. The maximum pressure during PACI was limited by the maximum insufflation pressure (30 mmHg) generated by the insufflator. Improvement of fetoscopic visualization as well as technical, maternal, and fetal safety aspects surrounding PACI were analyzed. RESULTS: PACI could successfully be instituted in 36 of the 37 procedures. In one case, when in the presence of increased uterine tone the opening pressure exceeded the maximum insufflation pressure of the insufflator, the strategy was abandoned. In all cases where PACI could be instituted successfully, the approach offered far superior visualization of the fetoscopic procedure than would have been possible within amniotic fluid. Acute or chronic maternal or fetal complications were observed in only one case (intraoperative membrane rupture). CONCLUSION: PACI greatly improves fetal visualization during fetoscopic interventions when fetoscopy within fluid meets with difficulties. Continued assessment of its benefits, risks, and safety margins at specialist centers is required. PMID- 19565299 TI - Telesimulation: an effective method for teaching the fundamentals of laparoscopic surgery in resource-restricted countries. AB - BACKGROUND: Several challenges exist with laparoscopic skills training in resource-restricted countries, including long travel distances required by mentors for onsite teaching. Telesimulation (TS) is a novel concept that uses the internet to link simulators between an instructor and a trainee in different locations. The purpose of this study was to determine the effectiveness of telesimulation for teaching the Fundamentals of Laparoscopic Surgery (FLS) to surgeons in Botswana, Africa. METHODS: A total of 16 surgeons from two centers in Botswana participated in this 8-week study. FLS TS was set up using two simulators, computers, webcams, and Skype software for eight surgeons in the TS group. A standard FLS simulator was available for the eight surgeons in the self practice (SP) group. Participants in the TS group had one remote training session per week with an FLS proctor at the University of Toronto who provided feedback and demonstrated proper technique. Participants in the SP group had access to the FLS DVD and were instructed to train on FLS at least once per week. FLS post-test scores were obtained in Botswana by a trained FLS proctor at the conclusion of the study. RESULTS: Participants in the TS group had significantly higher post test FLS scores than those in the SP group (440 +/- 56 vs. 272 +/- 95, p = 0.001). All trainees in the TS group achieved an FLS simulator certification passing score, whereas only 38% in the SP group did so (p = 0.03). CONCLUSION: Remote telesimulation is an effective method for teaching the Fundamentals of Laparoscopic Surgery in Africa, achieving a 100% FLS skills pass rate. This training platform provides a cost-effective method of teaching in resource restricted countries and could be used to teach laparoscopic skills anywhere in the world with internet access. PMID- 19565301 TI - Effective treatment of advanced colorectal cancer by rapamycin and 5 FU/oxaliplatin monitored by TIMP-1. AB - AIM: The mTOR-inhibitor rapamycin has shown antitumor activity in various tumors. Bedside observations have suggested that rapamycin may be effective as a treatment for colorectal carcinomatosis. METHODS: We established an orthotopic syngenic model by transplanting CT26 peritoneal tumors in Balb/C mice and an orthotopic xenograft model by transplanting SW620 peritoneal tumors in nu/nu mice. Expression levels of tissue inhibitor of matrix-metalloproteinases 1 (TIMP 1) in the tumor and serum was determined by enzyme-linked immunosorbent assay. RESULTS: Rapamycin significantly suppressed growth of syngenic and xenografted peritoneal tumors. The effect was similar with intraperitoneal or oral rapamycin administration. Tumor suppression was further enhanced when rapamycin was combined with 5-fluorouracil and/or oxaliplatin. The combination treatment showed no acute toxicity. TIMP-1 serum levels correlated well (CC = 0.75; P < 0.01) with rapamycin treatment. CONCLUSIONS: Rapamycin suppressed advanced stage colorectal cancer, even with oral administration. Combining rapamycin with current chemotherapy regimens significantly increased antitumor efficacy without apparent toxicity. The treatment efficacy correlated with serum TIMP-1 levels, suggesting its potential as a surrogate marker in future clinical trials. PMID- 19565302 TI - Establishment of a human neonatal hepatocyte cell line. AB - Hepatocytes are routinely used to generate and identify drug metabolites and hepatic toxicity. Primary cultures of human hepatocytes are the model cell of choice for most of these pharmacological and toxicological studies. However, major problems are encountered with primary liver cell cultures: the dwindling availability of viable livers, hepatocytes having a limited life span, the loss of liver-specific functions in culture, and the donor to donor variability. These limitations have created a significant need for an in vitro hepatocyte system, which has both the potential for use in toxicological and pharmaceutical studies as well as clinical applications. Ectopic expression of human telomerase reverse transcriptase (hTERT) is one of the major strategies used to develop immortalized cells. Immortalization of primary cells using hTERT allows retention of the original cellular characteristics and functions and avoids some of the genetic and phenotypic instabilities associated with using known oncogenes. In the present study, we developed a cell line from human neonatal hepatocytes by transduction with a recombinant retrovirus expressing the hTERT gene. Induction of stable expression of hTERT in the neonatal cells led to immortalization of these cells. The cell line was cultured continuously for more than 25 passages, equivalent to >25 population doublings, whereas the parental cells senesced within five passages. Analysis of telomerase activity as measured by telomeric repeat amplification protocol assay indicated elevated levels of telomerase activity in immortalized cells compared to the parental cells. These immortalized human hepatocytes cells maintained a normal diploid karyotype as well as the gene expression profile similar to that of human normal neonatal hepatocytes. The data suggest that these immortalized cells preserved some of the biological characteristics of hepatic progenitor cells and might be useful as an in vitro model for pharmacological and toxicity studies. PMID- 19565303 TI - Reporting results of orthopaedic research: confidence intervals and p values. PMID- 19565304 TI - Investigating the immunologic effects of CoCr nanoparticles. AB - The increase in metal-on-metal hip arthroplasties has led to concern regarding the effect of raised serum and tissue metal ion levels. Our aim was to determine changes in the integrity and function of cells of the immune system after exposure to CoCr nanoparticles in specific cell culture experiments. Nanometer sized particles of CoCr were made from a manufacturer's forged CoCr used for metal-on-metal articulations. Primary, murine dendritic cells and T and B lymphocytes then were exposed to these CoCr particles under cell culture conditions and then assayed for viability and proliferation/activation. CoCr nanoparticles did not directly activate dendritic cells or regulate B cells. Although nanoparticles were not directly toxic to resting T cells, Signals 1- and 2-dependent T cell proliferation were reduced. This may explain the observed reduction in CD8+ T cells observed in patients with metal-on-metal implants. PMID- 19565305 TI - Compress periprosthetic fractures: interface stability and ease of revision. AB - Periprosthetic fractures after massive endoprosthetic reconstructions pose a reconstructive challenge and jeopardize limb preservation. Compressive osseointegration technology offers the promise of relative ease of prosthetic revision, since fixation is achieved by means of a short intramedullary device. We retrospectively reviewed the charts of 221 patients who had Compress((R)) devices implanted in two centers between December, 1996 and December, 2008. The mean followup was 50 months (range, 1-123 months). Six patients (2.7%) sustained periprosthetic fractures and eight (3.6%) had nonperiprosthetic ipsilateral limb fractures occurring from 4 to 79 months postoperatively. All periprosthetic fractures occurred in patients with distal femoral implants (6/154, 3.9%). Surgery was performed in all six patients with periprosthetic femur fractures and for one with a nonperiprosthetic patellar fracture. The osseointegrated interface was radiographically stable in all 14 cases. All six patients with periprosthetic fracture underwent limb salvage procedures. Five patients had prosthetic revision; one patient who had internal fixation of the fracture ultimately underwent amputation for persistent infection. Periprosthetic fractures involving Compress((R)) fixation occur infrequently and most can be treated successfully with further surgery. When implant revision is needed, the bone preserved by virtue of using a shorter intramedullary Compress((R)) device as compared to conventional stems, allows for less complex surgery, making limb preservation more likely. LEVEL OF EVIDENCE: Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19565306 TI - Re: Bone graft harvest using a new intramedullary system. PMID- 19565308 TI - Direct determination of unbound lipophilic ligands in aqueous solutions. AB - Due to their hydrophobic nature, lipophilic compounds are always bound to proteins when transported in the organism. The transfer of such compounds between their binding proteins and cells as well as intracellular trafficking is mediated by a very low water-phase concentration of monomers. The use of protein filled resealed red cell membranes (erythrocyte ghosts) as semipermeable bags enables us to determine directly such water-phase concentrations in a biological system where the lipophilic compound is in equilibrium with the compound bound to its binding protein. Equilibrium dissociation constants (K(d)'s) and number of binding sites are determined by regression analyses of data. We describe the method with the hydrophobic anion arachidonate and the neutral N arachidonoylethanolamide as examples. PMID- 19565307 TI - Methylacridinium and its cholinergic properties. AB - 10-Methylacridinium iodide (methylacridinium; MA) is an inhibitor of cholinesterases. Inhibitors of acetylcholinesterase (AChE) are used in the treatment of myasthenia gravis, Alzheimer's disease, and in the prophylaxis of poisoning with organophosphates. Using spectrophotometric Ellman's method at 436 nm and commercial enzymes we found that MA inhibits AChE by binding with relatively high potency to the peripheral anionic site (IC(50) = 1.68 +/- 0.14 1M; human recombinant AChE) and equally to its inhibition of butyrylcholinesterase (BuChE; IC(50) = 3.54 +/- 0.27 1M; BuChE from human serum). MA also inhibits the binding of [(3)H]N-methylscopolamine to the muscarinic M2 receptor subtype, possibly in an allosteric manner (IC(50) = 1.90 1M). Functional effects on both the enzyme and the receptor could be observed in contractile studies on the isolated rat bladder. The ability of MA to cross the blood-brain barrier (log P = -0.32; polar surface area 3.88) provides prerequisites for a potential use of the drug in the treatment of neural disorders. PMID- 19565309 TI - Techniques of EMG signal analysis: detection, processing, classification and applications (Correction). AB - This paper was originally published in Biological Procedures Online (BPO) on March 23, 2006. It was brought to the attention of the journal and authors that reference 74 was incorrect. The original citation for reference 74, "Stanford V. Biosignals offer potential for direct interfaces and health monitoring. Pervasive Computing, IEEE 2004; 3(1):99-103." should read "Costanza E, Inverso SA, Allen R. 'Toward Subtle Intimate Interfaces for Mobile Devices Using an EMG Controller' in Proc CHI2005, April 2005, Portland, OR, USA." PMID- 19565310 TI - Analyzing folding and degradation of metabolically labelled polypeptides by conventional and diagonal sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - Efficient protein folding and quality control are essential for unperturbed cell viability. Defects in these processes may lead to production of aberrant polypeptides that are either degraded leading to "loss-of-function" phenotypes, or deposited in or outside cells leading to "gain-of-toxic-function" phenotypes. Elucidation of molecular mechanisms regulating folding and quality control of newly synthesized polypeptides is therefore of greatest interest. Here we describe protocols for metabolic labelling of transfected/infected mammalian cells with [(35)S]-methionine and [(35)S]-cysteine, for immunoisolation from detergent extracts of the selected model proteins and for the investigation of the model polypeptide's intracellular fate in response to chaperone-deletions or to cell exposure to folding or degradation inhibitors. PMID- 19565311 TI - The role of mast cells in non-ablative laser resurfacing with 1,320 nm neodymium:yttrium-aluminium-garnet laser. AB - The aim of this study was to investigate the role of mast cells in mechanisms of collagen remodelling induced by non-ablative laser treatment. The dorsal skin of Kunming (KM) mice was exposed to 1,320 nm neodymium-yttrium-aluminium garnet (Nd:YAG) laser weekly for four consecutive weeks. Biopsies were taken 1 h after irradiation and 1 day, 7 days, 14 days, 30 days and 60 days after the first treatment. Skin samples were studied for mast cells, fibroblasts, and type I and III collagen, by toluidine blue, haematoxylin-eosin (HE) and immunohistochemical staining, respectively. The total number of mast cells in the skin of experimental group was significantly greater than that in the control at 1 h, 1 day, 21 days and 60 days after the first treatment (P < 0.05, respectively). At any of the time points studied, the number of degranulated mast cells in the experimental group was significantly higher than in the control (P < 0.01, P < 0.01, P < 0.05, P < 0.01, P < 0.05, P < 0.05, respectively).The number of fibroblasts in the experimental group exhibited a significant increase in comparison with those in control skin at days 7, 21, 30 and 60 after irradiation (P < 0.05, P < 0.01, P < 0.01, P < 0.05, respectively). The amount of type I collagen was significantly higher than in the control from day 21 to day 60 (P < 0.05, P < 0.01, P < 0.01, respectively), and type III collagen showed a marked increase between day 7 and day 60, compared with the control (P < 0.01, P < 0.05, P < 0.01, P < 0.01, respectively). There was a significant positive correlation between the number of fibroblasts and granulated mast cells (r = 0.549, P < 0.01). The amount of type I and III collagen also showed significant positive correlations with the number of degranulated mast cells (r = 0.555, P < 0.01 and r = 0.579, P < 0.01, respectively). The results suggested that dermal mast cells might be involved in the inflammatory response, fibroblast proliferation and collagen remodelling induced by non-ablative laser treatment. PMID- 19565312 TI - Wavelength effect in temporomandibular joint pain: a clinical experience. AB - Temporomandibular disorders (TMDs) are common painful multifactorial conditions affecting the temporomandibular joint (TMJ) and whose treatment depends on the type and symptoms. Initially, it requires pain control, and, for this, drugs, biting plates, occlusal adjustment, physiotherapy or their association are used. Lately, laser phototherapy (LPT) has been used in the treatment of pain of several origins, including TMDs. This study reports the treatment of a selected group of 74 patients treated at the Laser Center of the Federal University of Bahia between 2003 and 2008. Following standard anamneses, clinical and imaging examination and with the diagnosis of any type of TMD, the patients were prepared for LPT. No other intervention was carried out during the treatment. Treatment consisted of three sessions a week for 6 weeks. Prior to irradiation, the patients were asked to score their pain using a visual analog scale (VAS). Lasers of wavelength (lambda)780 nm, lambda790 nm or lambda830 nm and/or lambda660 nm were used at each session (30/40 mW; spot (varphi) approximately 3 mm; mean dose per session 14.2 +/- 6.8 J/cm(2); mean treatment dose of 170 +/- 79.8 J/cm(2)). Of the patients, 80% were female ( approximately 46 years old). At the end of the 12 sessions the patients were again examined, and they scored their pain using the VAS. The results were statistically analyzed and showed that 64% of the patients were asymptomatic or had improved after treatment and that the association of both wavelengths was statistically significant (P = 0.02) in the asymptomatic group. It was concluded that the association of red and infrared (IR) laser light was effective in pain reduction on TMJ disorders of several origins. PMID- 19565313 TI - Short-course versus long-course neoadjuvant radiotherapy for lower rectal cancer: a systematic review. AB - OBJECTIVE: To analyse the efficacy of short-course (SCRT) versus long-course radiotherapy/chemoradiation (LCRT) as a neoadjuvant modality for the management of lower rectal cancer (LRC). METHODS: A systematic review of the literature was undertaken. Selected trials were analysed to generate a summative outcome. RESULTS: Seven trials on the efficacy of SCRT versus LCRT as a neoadjuvant modality for LRC encompassing 1,675 patients were retrieved but only 2 randomized trials on 396 patients qualified for this review. In both the fixed and random effects models, LCRT and SCRT were associated with equal overall survival, tumour recurrence, perioperative complications, sphincter preservation rate and toxicity. CONCLUSIONS: SCRT and LCRT may be as effective as traditional LCRT in terms of overall survival, recurrence, perioperative complications, sphincter preservation and toxicity. Traditional neoadjuvant chemoradiation may continue to be used. PMID- 19565314 TI - The incidence of acute stroke emergency admissions in an Irish teaching hospital. AB - BACKGROUND: Approximately 2,500 people die from stroke each year yet there is a lack of Irish services provision. AIMS: The aims of this study were to investigate the incidence of acute stroke emergency admissions in a large teaching hospital and present an analysis of this cohort. METHODS: All patients presenting acutely to the Emergency Department in 2005 meeting the WHO definition of stroke were included in our study. A chart review of the identified patients was conducted to obtain the necessary information. RESULTS: A total of 273 patients experienced an acute stroke, representing 1.6% of all acute admissions. 81.7% (223) of patients survived to discharge from the acute hospital. At 1 year, 65.2% (178) patients were still alive post-stroke. The mean length of stay in our acute hospital was 21.1 days following stroke. CONCLUSION: Stroke represents a considerable burden on health resources within the hospital. PMID- 19565315 TI - Usage of herbal medications in patients undergoing IVF treatment in an Irish infertility treatment unit. AB - BACKGROUND: Usage of herbal and complimentary medicines by patients presenting for procedures that require the use of anaesthetic agents is common. AIMS: To determine the prevalence of herbal medication usage among female patients attending for assisted reproduction procedures involving the use of sedative or hypnotic agents. METHODS: Questionnaire-based survey carried out over a 6-month period from January to July 2007. RESULTS: Forty-six percent of patients admitted regular use of herbal medications, with 38% of patients having taken herbal preparations in the 3-month period prior to their attendance for treatment. No patient taking herbal preparations had discussed the use of these products with the attending anaesthetist or fertility specialist prior to attending for the procedure. CONCLUSIONS: The usage of herbal medications with potential for serious adverse effects is high in this unique patient population. Education of both healthcare providers and patients is indicated. PMID- 19565316 TI - An audit of hip fracture services in the Mater Hospital Dublin 2001 compared with 2006. AB - BACKGROUND: Evidence-based practice supports the provision of integrated geriatric multidisciplinary care for older people with hip fractures. AIMS: To show that introduction of orthogeriatric services resulted in improved care and better patient outcomes. METHODS: We compared two audits: a 2001 audit of 103 patients over the age of 65 years with hip fracture and a 2006 audit on 98 admissions using the same parameters. RESULTS: Inpatient mortality dropped from 20% to 8%. Discharge to home increased from 8% to 18% and to nursing home fell from 25% to 21%. Prescription of perioperative antibiotics, DVT prophylaxis and osteoporosis medications improved. There were no significant differences in age or sex but there were significantly higher levels of disability in the 2001 group. CONCLUSION: Following introduction of an orthogeriatric service, improvements in mortality and discharge destination occurred. This may have been influenced by higher proportions of disability among the 2001 cohort. PMID- 19565317 TI - Pneumonitis caused by silicone gel following breast implant rupture. AB - INTRODUCTION: There have been reports of pneumonitis associated with subcutaneous injection of liquid silicone, and of other pulmonary conditions due to cohesive silicone gel prostheses, but we know of no previous cases of pneumonitis associated with silicone gel. MATERIALS AND METHODS: We report the case of a patient with a cohesive silicone gel mammary prosthesis in whom silicone-induced pneumonitis was diagnosed following radiological observation of pulmonary infiltrates and tests including transbronchial biopsy, which revealed the presence of silicone in alveolar histiocytes and small blood vessels. CONCLUSION: Following removal of the ruptured prosthesis and a course of systemic corticoids, the patient progressed favourably. PMID- 19565318 TI - Playing saxophone induced diffuse alveolar hemorrhage: a case report. AB - INTRODUCTION: The blood-gas barrier must be very thin to allow gas exchange and it is therefore subjected to high mechanical stresses when the capillary pressure rises. Exercise-induced pulmonary hemorrhage (EIPH) occurs frequently in horses and there is evidence that EIPH can also occur in humans. MATERIALS AND METHODS: We reported on a healthy 65-year-old male who developed a diffuse alveolar hemorrhage (DAH), like an EIPH, after playing saxophone for 6 h continuously. There were hemoptysis, crackles breathing sounds on exam, and bilateral radiographic infiltrates. A high-resolution computed tomographic study of the thorax disclosed DAH, the presence of which was proved by a gross appearance of bilateral bronchus on bronchoscopy and histopathological study of bronchoalveolar lavage material. CONCLUSION: This is the first report of alveolar hemorrhage caused by playing saxophone. In our case, he presented with a benign course and regressed spontaneously without any medical intervention. PMID- 19565319 TI - Adenosine A(2A) receptor gene (ADORA2A) variants may increase autistic symptoms and anxiety in autism spectrum disorder. AB - Autism spectrum disorders (ASDs) are heterogeneous disorders presenting with increased rates of anxiety. The adenosine A(2A) receptor gene (ADORA2A) is associated with panic disorder and is located on chromosome 22q11.23. Its gene product, the adenosine A(2A) receptor, is strongly expressed in the caudate nucleus, which also is involved in ASD. As autistic symptoms are increased in individuals with 22q11.2 deletion syndrome, and large 22q11.2 deletions and duplications have been observed in ASD individuals, in this study, 98 individuals with ASD and 234 control individuals were genotyped for eight single-nucleotide polymorphisms in ADORA2A. Nominal association with the disorder was observed for rs2236624-CC, and phenotypic variability in ASD symptoms was influenced by rs3761422, rs5751876 and rs35320474. In addition, association of ADORA2A variants with anxiety was replicated for individuals with ASD. Findings point toward a possible mediating role of ADORA2A variants on phenotypic expression in ASD that need to be replicated in a larger sample. PMID- 19565321 TI - Arcuate line of the rectus sheath: an anatomist's viewpoint. PMID- 19565320 TI - Clinical predictors of head-up tilt test outcome during the nitroglycerin phase. AB - OBJECTIVES: Nitrate-stimulated head-up tilt testing (HUT) is currently recommended to confirm the diagnosis of vasovagal syncope in subjects with syncope of unknown origin. Given the few data currently available, the aim of this study was to assess correlations between nitrate-induced HUT outcomes and the clinical characteristics of patients. METHODS: Two hundred and thirty consecutive, otherwise healthy subjects with a history of recurrent unexplained syncope underwent HUT. After 10 min supine rest, they were tilted to 70 degrees , and the test was potentiated by the administration of 300 microg of nitroglycerin after 20 min. RESULTS: Out of 178 subjects who underwent nitroglycerin administration during HUT, 95 fainted. At univariate Cox regression analysis, a reduced probability of VVS occurrence after nitrates was associated with greater systolic blood pressure and body mass index values, to male gender and smoking. At multivariate Cox regression analysis, only male gender (HR = 0.61; P = 0.039) and smoking (HR = 0.18; P = 0.001) remained significantly associated with HUT outcomes during the pharmacological phase of the test. INTERPRETATION: Smokers and males are less likely to faint after nitrate administration during HUT than non-smokers and females. Further studies should clarify the possibility of improving the diagnostic power of HUT in these patients. PMID- 19565322 TI - Passive mechanical properties of cardiac tissues in heart hypertrophy during pregnancy. AB - We evaluated changes in passive mechanical properties in cardiac tissues during rat pregnancy. Left and right ventricular free walls were dissected from hearts of nonpregnant, late-pregnant, and postpartum rats. Mechanical experiments in ventricular strips were done by stretch-release cycles using a step motor. The results show that during pregnancy, there is cardiac hypertrophy associated with (1) an increase in myocyte size, particularly of augmented myocyte length, (2) a decrease in passive tension developed by the myocardial walls, and (3) a decrease in both elastic modulus and hysteresis. All changes observed during rat pregnancy were reversed during postpartum. In conclusion, a heart with less ventricular rigidity could contribute to facilitating the ventricular filling in conditions of a greater circulating volume characteristic of pregnancy. PMID- 19565323 TI - Molecular and biological characterization of natural variants of Citrus dwarfing viroid. AB - Citrus dwarfing viroid has been proposed as an agent to control tree size in high density plantations. Thirty-three field isolates have been characterized, and the most frequent sequence/s have been identified. Five distinct variants were selected for biological characterization. Symptom expression analysis demonstrated a good correlation between leaf/stem symptoms and plant growth. The discriminating nucleotide sequence differences included two deletions and an insertion resulting in a reorganization of the base pairing of the terminal left loop, two (G42 --> A and C52 --> U) changes found in one of the variants, and as many as thirteen changes located in the right and left regions flanking the CCR. PMID- 19565325 TI - The fourth intracellular domain of G-protein coupling receptors: helicity, basicity and similarity to opsins. AB - The minimal size of the fourth intracellular domain of heptahelical G-protein coupling receptors (GPCRs) is close to 15 residues, and a juxtamembrane 15 residue segment is predicted as helical (Helix-8) in most of the receptors. Sequences of opsins, non-visual opsin-like (family A) GPCRs and Taste-2 receptors correspond with bovine rhodopsin at four positions in this tract. This is especially evident in monoamine receptors. In most GPCRs, the conserved juxtamembrane segment also has a large fraction of basic sidechains, and a considerable excess of cationic over anionic residues. The conservation is not dependent on the preferred G-protein alpha subunit or the overall length of the domain, indicating an additive speciation. In rod opsins and some A-GPCRs this segment has been shown to associate with the bilayer and to interact with G proteins. The segment could also be involved in precoupling of receptors and transducers. These interactions could be helped by both the structural propensities and the high content of cationic sidechains. PMID- 19565324 TI - Production and characterization of anti-dengue capsid antibodies suggesting the N terminus region covering the first 20 amino acids of dengue virus capsid protein is predominantly immunogenic in mice. AB - We produced monoclonal and polyclonal antibodies to the capsid (C) protein of dengue serotype 2 virus (DV2 C). First, a maltose-binding protein fused to DV2 C protein (MBP-C) was overproduced in E. coli. The affinity-purified MBP-C protein was cleaved by factor Xa protease to obtain a recombinant DV2 C protein, which was then used for mouse immunizations. Two hybridoma cell lines producing anti-C Mabs as well as anti-C polyclonal antibody were successfully generated and characterized. Interestingly, all of the generated antibodies specifically recognized the first 20 amino acids of the DV2 C protein, as determined by peptide epitope mapping and via a recombinant DV2 C protein in which this region was deleted. The results suggested that this region is predominantly immunogenic in mice. PMID- 19565326 TI - NMDA receptor activation induces differential epigenetic modification of Bdnf promoters in hippocampal neurons. AB - Transcriptional regulation of the gene encoding brain-derived neurotrophic factor (BDNF) has been widely studied. However, an understanding of mechanisms modifying chromatin, events that are essential for controlling transcription, is rudimentary. We focused on two activation-dependent regions of the Bdnf gene physically linked to known transcription sites for exons 1 and 4. Using chromatin immunoprecipitation assays, we determined that N-methyl-D-aspartate (NMDA) receptor activation derepressed promoters 1 and 4-mediated transcription. This derepression correlated with reduced occupancy by histone deacetylase 1 and methyl cytosine-binding protein 2 of each promoter region near known transcription start sites in cultured hippocampal neurons. These changes did not occur at all sites upstream of transcription initiation. Taken together, these findings suggest that histone and other DNA-binding proteins are involved in remodeling of chromatin at some, but not all sites, within Bdnf promoters 1 and 4 and are associated with NMDA receptor-dependent increases in transcription. PMID- 19565327 TI - PTSD and postpartum mental health in a sample of Caucasian, Asian, and Pacific Islander women. AB - To better understand the role of posttraumatic stress disorder (PTSD) in postpartum health, this study investigates the relationship of PTSD and associated perinatal behavioral risk factors in a sample of Caucasian, Asian, and Pacific Islander women. As part of a larger longitudinal study, 54 women (18-35 years of age) were interviewed at their postpartum clinic visit for PTSD, anxiety, depression, and alcohol and substance use. PTSD and subclinical PTSD during the postpartum period were associated with behavioral health risks, with PTSD at the onset of pregnancy being a predictor of postpartum PTSD by a factor of three. Women with PTSD and subclinical PTSD were more likely to also experience stress (73%), anxiety (64%), and depression (73%) during the postpartum period compared to those without PTSD. No significant differences were found by ethnicity for postpartum PTSD, depression, or anxiety. Regardless of ethnicity or PTSD status, one in four women in the sample had a probable mental health disorder or risk behavior of some type during the postpartum period. Given the rates of associated mental health risks with PTSD, these findings suggest further research examining the fluctuations of PTSD symptomatology throughout each pregnancy trimester to determine its role as a potential mediator during the perinatal period. Further research is also needed to elucidate the role of ethnic or cultural differences in trauma and PTSD and perinatal health. PMID- 19565328 TI - Depressive and anxiety symptoms through late pregnancy and the first year post birth: an examination of prospective relationships. AB - One hundred and fifty-nine women were measured for depressive and anxiety symptoms from late pregnancy through to 12 months postpartum. Partial correlations revealed stability of depressive and anxiety symptoms across time. Depressive symptoms did not predict anxiety at any time point. Anxiety predicted increases in depressive symptoms from late pregnancy to early postpartum, but not from early postpartum to mid postpartum. Anxiety predicted depressive symptoms from mid postpartum to late postpartum, however, not when social support in late pregnancy was controlled for. PMID- 19565329 TI - Sequence dependent femtosecond-resolved hydration dynamics in the minor groove of DNA and histone-DNA complexes. AB - Understanding the sequence dependent molecular recognition of DNA is crucial for the rational design of many drugs. Femtosecond resolved studies on the hydration dynamics of the dodecamer duplexes having sequences (CGCGAATTCGCG)2 and (CGCAAATTTGCG)2, and their complexes with the nucleic protein histone 1 (H1) reveal significant correlation of the molecular recognition of the DNA and DNA protein complexes with the dynamics of hydration. The different molecular recognition of DNA and DNA-protein complexes is also borne out by circular dichroism (CD) and fluorescence detected CD measurements. PMID- 19565331 TI - Brief report: Further evidence for inner speech deficits in autism spectrum disorders. AB - Recent research indicates that individuals with autism do not effectively use inner speech during the completion of cognitive tasks. We used Articulatory Suppression (AS) to interfere with inner speech during completion of alternate items from the Tower of London (TOL). AS detrimentally affected TOL performance among typically developing (TD) adolescents (n = 25), but did not significantly diminish performance among adolescents with high functioning (IQ > 80) autism spectrum disorders (n = 28). Moreover, the TD group's TOL performance under AS was indistinguishable from the autism group's impaired baseline TOL performance. These findings suggest that diminished inner speech usage among individuals with high functioning autism spectrum disorders (relative to TD controls) may contribute to executive dysfunction associated with these disorders. PMID- 19565332 TI - Parvovirus H-1 induces cytopathic effects in breast carcinoma-derived cultures. AB - Parvovirus H-1 (H-1 PV) preferentially replicates in malignant cells resulting in their death by cytolysis. It has often been considered a potential candidate for use in novel anticancer therapy. To evaluate its potential in a model of natural tumors, we assayed in vitro the effect exerted by H-1 PV on short-term cultures derived from breast tumor samples freshly excised from patients. Our results show that H-1 PV effectively kills tumor-derived cells, whereas normal tissue-derived cells showed no H-1 PV-induced cytopathic effects (CPE). We also determined that the H-1 PV sensitivity (up to 67% sensitive cultures) is related with the quantities of virus assayed. We further examined the expression and phosphorylation state of the parvoviral nonstructural protein 1 (NS1), known to be associated with parvoviruses-induced CPE. Both appear to be impaired in normal tissue-derived cells and resistant cultures. Finally, we show that H-1 PV sensitivity in cultures correlates significantly with higher tumor grades (Nottingham combined histologic grade 2 or 3). This report confirms that H-1 PV can efficiently induce CPE in primary breast tumor cells in vitro. It identifies tumor characteristics representing potential criteria for recruiting patients for clinical evaluation of H-1 PV antitumor effects. PMID- 19565330 TI - Patterns of methamphetamine use during pregnancy: results from the Infant Development, Environment, and Lifestyle (IDEAL) Study. AB - The objectives of this study are to characterize methamphetamine (MA) usage patterns during pregnancy, examine whether patterns of MA use are associated with sociodemographic characteristics and prenatal care, and test the hypothesis that persistent or increasing MA use during pregnancy is associated with greater use of other illicit drugs. The sample consisted of 191 MA-using mothers who participated in a large-scale multi-site study of prenatal MA exposure. Patterns of substance use were assessed by maternal self-report via the Substance Use Inventory (SUI), which included detailed information about MA use, including frequency, quantity, and maximum use during each trimester of pregnancy. The study demonstrated that on average, the prevalence of MA use decreased over the three trimesters of pregnancy (84.3% vs. 56.0% vs. 42.4%), and decreased frequency was observed among users from the first trimester to the third (3.1 vs. 2.4 vs. 1.5 days/week). Closer examination of the individual patterns revealed that 29.3% of women maintained consistently high frequency, 9.4% increased frequency, 25.7% had a stable low/moderate pattern, and 35.6% decreased their frequency of MA over the course of pregnancy. These four groups did not differ in sociodemographic characteristics; women who decreased their use of MA had significantly more prenatal visits compared to the consistently high-use group, but were the most likely to use alcohol during their pregnancy. In conclusion, this article elucidated the different patterns of MA use in this community sample. Approximately, one third of MA-using mothers could be classified as consistently high users with a profile of use with the greatest risk to themselves and potentially to their infants including high levels of MA use throughout pregnancy and fewer prenatal care visits. Overall, we found that MA use declined across pregnancy; however, a substantial proportion of users had consistently high or increasing MA use, while those who decreased their MA frequency had a higher prevalence of polydrug use. Future research will investigate the association of these patterns with neonatal outcomes. PMID- 19565333 TI - Dietary beta-carotene, vitamin C and E intake and breast cancer risk in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - So far, studies on dietary antioxidant intake, including beta-carotene, vitamin C and vitamin E, and breast cancer risk are inconclusive. Thus, we addressed this question in the European Prospective Investigation into Cancer and Nutrition. During a median follow-up time of 8.8 years, 7,502 primary invasive breast cancer cases were identified. Cox proportional hazard models were used to estimate hazard ratios (HR) and 95% confidence intervals (CI). All analyses were run stratified by menopausal status at recruitment and, additionally, by smoking status, alcohol intake, use of exogenous hormones and use of dietary supplements. In the multivariate analyses, dietary intake of beta-carotene, vitamin C and E was not associated with breast cancer risk in premenopausal [highest vs. lowest quintile: HR, 1.04 (95% CI, 0.85-1.27), 1.12 (0.92-1.36) and 1.11 (0.84-1.46), respectively] and postmenopausal women [0.93 (0.82-1.04), 0.98 (0.87-1.11) and 0.92 (0.77-1.11), respectively]. However, in postmenopausal women using exogenous hormones, high intake of beta-carotene [highest vs. lowest quintile; HR 0.79 (95% CI, 0.66-0.96), P (trend) 0.06] and vitamin C [0.88 (0.72-1.07), P (trend) 0.05] was associated with reduced breast cancer risk. In addition, dietary beta carotene was associated with a decreased risk in postmenopausal women with high alcohol intake. Overall, dietary intake of beta-carotene, vitamin C and E was not related to breast cancer risk in neither pre- nor postmenopausal women. However, in subgroups of postmenopausal women, a weak protective effect between beta carotene and vitamin E from food and breast cancer risk cannot be excluded. PMID- 19565334 TI - The association of religiosity, sexual education, and parental factors with risky sexual behaviors among adolescents and young adults. AB - This study examined the association of religiosity, sexual education and family structure with risky sexual behaviors among adolescents and young adults. The nationally representative sample, from the 2002 National Survey of Family Growth, included 3,168 women and men ages 15-21 years. Those who viewed religion as very important, had frequent church attendance, and held religious sexual attitudes were 27-54% less likely to have had sex and had significantly fewer sex partners than peers. Participants whose formal and parental sexual education included abstinence and those from two-parent families were 15% less likely to have had sex and had fewer partners. PMID- 19565335 TI - The role and extent of pelvic lymphadenectomy in the management of patients with invasive urothelial carcinoma. AB - Lymph node metastases are the most important prognostic variable in determining outcome following radical cystectomy. An anatomic bilateral node dissection includes at a minimum the external and internal iliac and obturator lymph nodes. An extended node dissection may include the distal aortic and vena caval nodes, bilateral common iliac, and pre-sacral nodes, which receive direct lymphatic drainage from the posterior bladder and trigone. This approach sets up the cystectomy, maximizes sensitivity for detection of nodal metastasis, assures optimum local pelvic cancer control, and accurately identifies those high-risk patients with node metastases who may benefit from adjuvant chemotherapy. Lymph node retrieval is affected by several variables of node specimens addition to the anatomic extent of the node dissection. These include presentation to the pathologist in packets, specimen processing and what the pathologist calls a lymph node, and patient age. The current TNM staging system accounts for the number and size of node metastases and may be improved by incorporating lymph node density, which is a composite variable incorporating the number of positive nodes and number of nodes retrieved--a possible surrogate for the extent of the node dissection. Innovations in imaging including novel MRI contrast agents and lymphoscintigraphy may improve the pre-treatment and intra-operative identification of node metastases and lymphatic anatomy. Minimally invasive surgical techniques including robotic-assisted laparoscopic cystectomy may improve peri-operative outcomes but must meet the standard of anatomic node dissection and long-term cancer control afforded by the gold standard of anatomic radical cystectomy and bilateral pelvic and iliac node dissection. PMID- 19565336 TI - Challenges of fragment screening. PMID- 19565337 TI - Millisecond dynamics in the allosteric enzyme imidazole glycerol phosphate synthase (IGPS) from Thermotoga maritima. AB - IGPS is a 51 kDa heterodimeric enzyme comprised of two proteins, HisH and HisF, that catalyze the hydrolysis of glutamine to produce NH(3) in the HisH active site and the cyclization of ammonia with N'-[(5'-phosphoribulosyl)formimino]-5 aminoimidazole-4-carboxamide-ribonucleotide (PRFAR) in HisF to produce imidazole glycerol phosphate (IGP) and 5-aminoimidazole-4-carboxamide ribotide (AICAR). Binding of PRFAR and IGP stimulates glutaminase activity in the HisH enzyme over 5,000 and 100-fold, respectively, despite the active sites being >25 A apart. The details of this long-range protein communication process were investigated by solution NMR spectroscopy and CPMG relaxation dispersion experiments. Formation of the heterodimer enzyme results in a reduction in millisecond motions in HisF that extend throughout the protein. Binding of lGP results in an increase in protein-wide millisecond dynamics evidenced as severe NMR line broadening and elevated R (ex) values. Together, these data demonstrate a grouping of flexible residues that link the HisF active site with the protein interface to which HisH binds and provide a model for the path of communication between the IGPS active sites. PMID- 19565338 TI - Biochemical characterization of Helicobacter pylori alpha-1,4 fucosyltransferase: metal ion requirement, donor substrate specificity and organic solvent stability. AB - The effect of metal ions on the activity, the donor substrate specificity, and the stability in organic solvents of Helicobacter pylori alpha-1,4 fucosyltransferase were studied. The recombinant enzyme was expressed as soluble form in E. coli strain AD494 and purified in a one step affinity chromatography. Its activity was highest in cacodylate buffer at pH 6.5 in the presence of 20 mM Mn2+ ions at 37 degrees C. Mn2+ ions could be substituted by other metal ions. In all cases, Mn2+ ions proofed to be the most effective (Mn2+ > Co2+ > Ca2+ > Mg2+ > Cu2+ > Ni2+ > EDTA). The enzyme shows substrate specificity for Type I disaccharide (1) with a KM of 114 MUM. In addition, the H. pylori alpha-1,4 fucosyltransferase efficiently transfers GDP-activated L-fucose derivatives to Galbeta1-3GlcNAc-OR (1). Interestingly, the presence of organic solvents such as DMSO and methanol up to 20% in the reaction medium does not affect significantly the enzyme activity. However, at the same concentration of dioxane, activity is totally abolished. PMID- 19565339 TI - Allelic loss at TP53 in metastatic human endometrial carcinomas. AB - Loss of heterozygosity (LOH) is implicated in the initiation and progression of various human neoplasia, and is observed in both early or in advanced-stage human cancers. The current study was aimed at investigating the frequency of LOH TP53 in human endometrial carcinoma (EC) metastases. LOH was analyzed using 3 intragenic polymorphisms in 38 primary ECs and corresponding metastatic lesions. Allelic loss at intron 1 was detected in 14 out of 38 (37%) primary carcinomas and in 11 out of 38 (29%) metastatic lesions. LOH at intron 1 in primary and metastatic tumors was concomitantly noted in 8 out of 38 (21%) cases. LOH at intron 4 was seen in 46% (17 out of 37) primary ECs and in 35% (13 out of 37) metastatic lesions. LOH at intron 4 in primary tumor/metastasis was concomitantly demonstrated in 27% (10 out of 33) cases. Allelic loss at exon 4 was detected in 5 out of 33 (15%) primary ECs and in one out of 33 (3%) corresponding metastases. Coexistence of LOH TP53 in primary ECs with metastases at intron 1 and intron 4 was observed in three out of 33 (9%) cases. Correlation between allelic loss at intron 1 in primary ECs and corresponding metastases was found (R = 0.475, p = 0.003). Moreover, there was correlation between LOH at intron 1 in metastastic ECs and allelic imbalance at intron 4 in primary uterine tumors (R = 0.416, p = 0.01). There was a relationship between LOH at intron 4 in primary ECs and corresponding metastatic lesions (R = 0.457, p = 0.004). LOH TP53 at intron 4 correlated with the presence of the neoplasm in the uterine cervix (R = 0.319, p = 0.049), and with the non-endometrioid type of primary tumor (R = 0.371, p = 0.024). There was a significant correlation between exon 4 LOH and patient age (less or equal to 50 years and above this age; R = -0.375, p = 0.032). p53 overexpression was present in thirteen out of 38 (34%) cases, both in primary ECs and in metastatic lesions. Overexpression of p53 was higher in non-endometrioid ECs (three out of 5; 60%) than in endometrioid-type uterine tumors (ten out of 33; 30.3%; p = 0.315). p53 overexpression correlated with the presence of cancer in the lumen of fallopian tube(s) (R = 0.032, p = 0.046), and with allelic loss at intron 1 in primary ECs (R = 0.599, p = 0.0001). In conclusion, LOH occurs not only in primary uterine tumors but also in corresponding metastases, with the higher incidence being reported at intron 4 of the TP53. A significant link existed between LOH TP53 at intron 1 and p53 overexpression in primary ECs, but not in the corresponding metastatic lesions. PMID- 19565340 TI - EEG sources of noise in intraoperative somatosensory evoked potential monitoring during propofol anesthesia. AB - OBJECTIVE: It was hypothesized that somato- sensory evoked potentials can be achieved faster by selective averaging during periods of low spontaneous electroen- cephalographic (EEG) activity. We analyzed the components of EEG that decrease the signal-to-noise ratio of somatosensory evoked potential (SEP) recordings during propofol anesthesia. METHODS: Patient EEGs were recorded with a high sampling frequency during deep anesthesia, when EEGs were in burst suppression. EEGs were segmented visually into bursts, spindles, suppressions, and artifacts. Tibial somatosensory evoked potentials (tSEPs) were averaged offline separately for burst, suppression, and spindle segments using a signal bandwidth of 30-200 Hz. Averages achieved with 2, 4, 8, 16, 64, 128, and 256 responses were compared both visually, and by calculating the signal-to-noise ratios. RESULTS: During bursts and spindles, the noise levels were similar and significantly higher than during suppressions. Four to eight times more responses had to be averaged during bursts and spindles than during suppressions in order to achieve a similar response quality. Averaging selectively during suppressions can therefore yield reliable tSEPs in approximately one-fifth of the time required during bursts. CONCLUSION: The major source of EEG noise in tSEP recordings is the mixed frequency activity of the slow waves of bursts that occur during propofol anesthesia. Spindles also have frequency components that increase noise levels, but these are less important, as the number of spindles is fewer. The fastest way to obtain reliable tSEPs is by averaging selectively during suppressions. PMID- 19565341 TI - Dietary change among breast and colorectal cancer survivors and cancer-free women in the Norwegian Women and Cancer cohort study. AB - OBJECTIVE: To study diet before and after diagnosis of breast and colorectal cancers compared with diet in cancer-free women in the Norwegian Women and Cancer study. METHODS: This paper reports dietary changes from a data collection in 1996 1999 to another in 2002-2005. A total of 43,847 cancer-free women aged 41-70 years answered the baseline questionnaire on diet and lifestyle, 130 women developed colorectal cancer and 563 breast cancer. Dietary change in the three groups was compared, for breast cancer a comparison was made according to stage and time since diagnosis. RESULTS: Breast cancer survivors increased fruit and vegetable consumption with 81 g compared to 42 g in colorectal cancer survivors and 50 g in cancer-free women (p difference in change <0.001). Milk consumption decreased among cancer-free women, but not among colorectal cancer survivors (p = 0.007). Significantly more cancer survivors quit smoking (p < 0.001). There were no differences in change of alcohol consumption or BMI. In breast cancer survivors, differences increased with time since diagnosis, and stage II survivors made larger changes than stage I survivors. CONCLUSIONS: Cancer survivors showed little change toward cancer-preventive guidelines, although more advanced stage and being more than 2.4 years post diagnosis was associated with greater change in diet and smoking behaviors. PMID- 19565342 TI - Timing of births and endometrial cancer risk in Swedish women. AB - While a protective long-term effect of parity on endometrial cancer risk is well established, the impact of timing of births is not fully understood. We examined the relationship between endometrial cancer risk and reproductive characteristics in a population-based cohort of 2,674,465 Swedish women, 20-72 years of age. During follow-up from 1973 to 2004, 7,386 endometrial cancers were observed. Compared to uniparous women, nulliparous women had a significantly elevated endometrial cancer risk (hazard ratio [HR] = 1.32, 95% confidence interval [CI], 1.22-1.42). Endometrial cancer risk decreased with increasing parity; compared to uniparous women, women with > or =4 births had a HR = 0.66 (95% CI, 0.59-0.74); p trend < 0.001. Among multiparous women, we observed no relationship of risk with age at first birth after adjustment for other reproductive factors. While we initially observed a decreased risk with later ages at last birth, this appeared to reflect a stronger relationship with time since last birth, with women with shorter times being at lowest risk. In models for multiparous women that included number of births, age at first and last birth, and time since last birth, age at last birth was not associated with endometrial cancer risk, while shorter time since last birth and increased parity were associated with statistically significantly reduced endometrial cancer risks. The HR was 3.95 (95% CI; 2.17 7.20; p-trend = <0.0001) for women with > or =25 years since a last birth compared to women having given birth within 4 years. Our findings support that clearance of initiated cells during delivery may be important in endometrial carcinogenesis. PMID- 19565343 TI - Quantitative analysis of shoreline changes at the Mediterranean Coast in Turkey. AB - This research is focused on the coastline evolution monitoring and its potential change estimation by remote sensing techniques using multi-temporal Landsat images at the southeast coasts of the Mediterranean Sea in Turkey. The study area includes the coastal zone located in the Cukurova Delta coasts. The Cukurova Delta has accreted toward the Mediterranean Sea as a result of sediment discharge and transport from Seyhan and Ceyhan rivers. These processes have caused the morphological changes (accretion or erosion) of coastline along some parts of the southeast coasts of the Mediterranean Sea. In this study, coastline changes were researched by using radiometrically and geometrically corrected multi-temporal and multi-spectral data from Landsat Multispectral Scanner dated 1972, Thematic Mapper dated 1987, and Enhanced Thematic Mapper dated 2002. In the image processing steps, mosaicing, subset, Iterative Self-Organizing Data Analysis Technique classification, band ratioing (B5/B2), edge detection, and overlay techniques were used to carry out coastline extraction and the Digital Shoreline Analysis System was used to calculate rate of coastline changes. As a result of the analysis, in some parts of the research area, remarkable coastline changes (more than 2,900 m withdrawal and -24.50 m/year erosion) were observed for a 30 year period. PMID- 19565344 TI - Environmental change monitoring in the arid and semi-arid regions: a case study Al-Basrah Province, Iraq. AB - In recent years, land use/cover dynamic change has become a key subject urgently to be dealt with in the study of global environmental change. This research utilizes the integrated remote sensing and geographic information systems (GIS) in the southern part of Iraq (Basrah Province was taken as a case) to monitor, map, and quantify the environmental change using a 1:250,000 mapping scale. Remote sensing and GIS software were used to classify Landsat TM in 1990 and Landsat ETM+ in 2003 imagery into five land use and land cover (LULC) classes: vegetation land, sand land, urban area, unused land, and water bodies. Supervised classification and normalized difference buildup index, normalized difference vegetation index, normalized difference bare land index, the normalized differential water index, crust index (CI) algorithms, and change detection techniques were adopted in this research and used, respectively, to retrieve its class boundary. An accuracy assessment was performed on the 2003 LULC map to determine the reliability of the map. Finally, GIS software was used to quantify and illustrate the various LULC conversions that took place over the 13-year span of time. The results showed that the urban area, sand lands, and bare lands had increased by the rate of 1.2%, 0.8%, and 0.4% per year, with area expansion from 3,299.1, 4,119.1 km2, and 3,201.9 km2 in 1990 to 3,794.9, 4,557.7, and 3,351.7 km2 in 2003, respectively. While the vegetation cover and water body classes were about 43.5% in 1990, the percentage decreased to about 39.6% in 2003. This study demonstrates the effectiveness of the remote sensing and GIS technologies in detecting, assessing, mapping, and monitoring the environmental changes. PMID- 19565345 TI - Heavy metal concentrations in water, suspended matter, and sediment from Gokova Bay, Turkey. AB - The contents of heavy metals (Fe, Mn, Pb, Cu, Cd, and Hg) dissolved in water and suspended solids of Gokova Bay--partly and fully sampled in 2005 and 2006, respectively--are quite higher than the average values encountered in uncontaminated sea water. The high concentrations are associated with terrestrial inputs from the mining zones and anthropogenic (domestic+industrial) sources. Moreover, the distribution of Fe and Cu is affected by primary production because these elements function as nutrients in biological activities. The Cr, Ni, and Fe concentrations of surface sediments are above the shale average. The Cr and Ni contents of surface sediments representative of river mouths strongly correlate with total phosphorus contents. In a sulfide-poor environment, Pb and Cu were concentrated at a higher ratio in surface sediments than Cd, probably due to higher stabilities of their surface complexes with amorphous iron oxides and clay minerals existing as major components in the sediments. The exceptional enrichment of Zn may be attributed to double oxide formation with amorphous iron oxides in sediments. The high metal values are most probably caused by terrestrial inputs from anthropogenic sources and the mining zones at the southeast part of the bay. The Al, Mn, Pb, Cu, Zn, and Hg contents are below the shale average. The low values have possibly originated from the coarse-grained sandy sediments having a low affinity for metals. There are no distinct differences in the metal distributions in water and suspended matter between the years 2005 and 2006 in the bay, probably due to low sedimentation rates. PMID- 19565346 TI - Water chemistry influences the toxicity of silver to the green-lipped mussel Perna viridis. AB - The study determined the influence and relative importance of water chemistry parameters (pH, alkalinity, hardness) on the acute toxicity of silver to the green mussel Perna viridis. A preliminary bioassay revealed that 4 mg L(-1) of silver caused 50% mortality (LC50) in 96 h for mussels placed in seawater with pH 8.5, hardness 1,872 mg L(-1), and alkalinity 172 mg L(-1). Mortality of mussels increased with decreasing pH and increasing hardness and alkalinity variables. In contrast the mortality decreased with increasing pH and decreasing hardness and alkalinity values. The water chemistry also affected the concentration of silver in experimental seawater and bioaccumulation of silver in mussels. The results revealed that the chemical properties of seawater must be considered while conducting toxicity tests with metals like silver. The possible explanations for the influence of water chemistry on silver toxicity to P. viridis are discussed. PMID- 19565347 TI - Determination of total Cr in wastewaters of Cr electroplating factories in the I.organize industry region (Kayseri, Turkey) by ICP-AES. AB - The wastewater pollution in industrial areas is one of the most important environmental problems. Heavy metal pollution, especially chromium pollution in the wastewater sources from electroplating, dyeing, and tannery, has affected the life on earth. This pollution can affect on all ecosystems and human health directly or by food chain. Therefore, the determination of total chromium in this study is of great importance. In this study, accurate, rapid, sensitive, selective, simple, and low-cost technique for the direct determination of total Cr in wastewater samples collected from the some Cr electroplating factories in March 2008 by inductively coupled plasma-atomic emission spectrometry has been developed. The analysis of a given sample is completed in about 15 min by this technique applied. As the result of the chromium analysis, the limit of quantification for the total Cr were founded to be over the limit value (0.05 mg L(-1); WHO, EPA, TSE 266, and inland water quality classification) as 1,898.78+/ 0.34 mg/L at station 1 and 3,189.02+/-0.56 mg/L at station 2. The found concentration of total Cr has been determined to be IV class quality water according to the inland water classification. In order to validate the applied method, recovery studies were performed. PMID- 19565348 TI - Detection of the Maedi Visna Virus in the popliteal lymph nodes of sheep infected by the respiratory route. PMID- 19565349 TI - A case of "pseudo science"? A study claiming effects of the Cry1Ab protein on larvae of the two-spotted ladybird is reminiscent of the case of the green lacewing. PMID- 19565350 TI - Hydrocarbon degradation by Dietzia sp. A14101 isolated from an oil reservoir model column. AB - The hydrocarbon-degrading strain Dietzia sp. A14101 was isolated from an oil reservoir model column inoculated with oil-field bacteria. The column was continuously injected with nitrate (0.5 mM) from the start of water flooding, which lead to a gradual development of nitrate reduction in the column. Strain A14101 was able to utilize a range of aliphatic hydrocarbons as sole carbon and energy source during aerobic growth. Whole oil gas chromatography analysis of the crude oil phase from aerobic pure cultures showed that strain A14101 utilized the near complete range of aliphatic components and aromatic components toluene and xylene. Longer n-alkanes >/=C(17) were utilized simultaneously with the shorter C(10) and C(15). After 120 days aerobic incubation, the whole oil gas chromatography profile of the crude oil phase was similar to that of heavily biodegraded oils. Anaerobic degradation of hydrocarbons with nitrate was not observed. Nitrate reduction was, however, observed during anaerobic growth on propionate, which suggests that strain A14101 grows on fatty acids in the column rather than on hydrocarbons. PMID- 19565351 TI - Virulotyping of Salmonella enterica subsp. enterica isolated from indigenous vegetables and poultry meat in Malaysia using multiplex-PCR. AB - The increased occurrence of Salmonella occurrence in local indigenous vegetables and poultry meat can be a potential health hazards. This study is aimed to detect the prevalence of twenty different virulence factors among Salmonella enterica strains isolated from poultry and local indigenous vegetables in Malaysia via an optimized, rapid and specific multiplex PCR assay. The assay encompasses a total of 19 Salmonella pathogenicity islands genes and a quorum sensing gene (sdiA) in three multiplex reaction sets. A total of 114 Salmonella enterica isolates belonging to 38 different serovars were tested. Each isolate in under this study was found to possess up to 70% of the virulence genes tested and exhibited variable pathogenicity gene patterns. Reproducibility of the multiplex PCR assay was found to be 100% and the detection limit of the optimized multiplex PCR was tested with lowest detectable concentration of DNA 0.8 pg microl(-1). This study demonstrated various Salmonella pathogenicity island virulence gene patterns even within the same serovar. This sets of multiplex PCR system provide a fast and reliable typing approach based on Salmonella pathogenicity islands, thus enabling an effective monitoring of emerging pathogenic Salmonella strains as an additional tool in Salmonella surveillance studies. PMID- 19565352 TI - Association between Fc receptor-like 3 C169T polymorphism and risk of systemic lupus erythematosus: a meta-analysis. AB - To investigate the association between Fc receptor-like 3 (FCRL3) C169T polymorphism and susceptibility to systemic lupus erythematosus (SLE). We surveyed studies on the FCRL3 C169T polymorphism and SLE using comprehensive Medline search and review of the references. Meta-analysis was performed for genotypes CC (recessive effect), CC+CT (dominant effect) and C allele in fixed effects model or random effects model. Five identified studies included 1,944 SLE patients and 4,528 non-SLE controls. Three out of five identified studies included populations of Asian descent, and two included populations of European descent. The overall odds ratio (OR) of the CC genotype was 1.21(95% confidence interval [CI], 1.04-1.40). Stratification by ethnicity indicated that the CC genotype was associated with SLE in Asian-derived population (OR, 1.23; 95% CI, 1.03-1.47). No association was detected in European-derived population (OR, 1.17; 95% CI, 0.90-1.52). This meta-analysis fails to show significant association of CC+CT genotypes and C allele with SLE in overall, European-derived and Asian derived populations. In summary, this meta-analysis demonstrates that the FCRL3 169CC genotype (recessive effect) may confer susceptibility to SLE, especially in Asian-derived population. PMID- 19565353 TI - Duplex (or quadruplet) CH domain containing human multidomain proteins: an inventory. AB - In this paper, the inventory presented for singlet CH (calponin homology/actin binding) domain containing human multidomain proteins is extended to several duplex and one quadruplet CH containing forms. Invariably, the duplexes are located at the begin of the molecules. The regions connecting the two CH units suggest amino acid conservations which allows the placing of 18 duplex containing molecules into six groups wherein the gene for one member in each group created the others more recently by gene duplication. The ancient multidomain proteins, possibly, were primarily the result of an exon shuffling (transposition) mechanism that also guided the placing of the CH singlet or duplex domain at the amino end of the newly created proteins. A mechanism that creates pseudogenes could conceivably produce genes that encode multi-domain proteins. Intragenomic duplications (slippage) might have facilitated the occurrence of encoding repeats, thus allowing for the creation of multiple identical domains within one molecule. Gene duplication with subsequent modification and small domain gene recombination which formed multidomain proteins are important forces driving evolution. PMID- 19565355 TI - In vitro culture and differentiation of osteoblasts from human umbilical cord blood. AB - It is well accepted that human umbilical cord blood (UCB) is a source of mesenchymal stem cells (MSCs) which are able to differentiate into different cell phenotypes such as osteoblasts, chondrocytes, adipocytes, myocytes, cardiomyocytes and neurons. The aim of this study was to isolate MSCs from human UCB to determine their osteogenic potential by using different kinds of osteogenic medium. Eventually, only those MSCs cultured in osteogenic media enriched with vitamin D(2) and FGF9, were positive for osteocalcin by RT-PCR. All these cells were positive for alizarin red, alkaline phosphatase and Von Kossa. The results obtained from RT-PCR have confirmed that osteogenesis is complete by expression of the osteocalcin marker. In conclusion, vitamin D(2), at least in vitro, may replace vitamin D(3) as an osteogenic stimulator factor for MSC differentiation. PMID- 19565356 TI - The role of p38alpha mitogen-activated protein kinase gene in the HELLP syndrome. AB - Mitogen-activated protein kinase (MAPK) p38alpha was shown to be implicated in the organogenesis of the placenta, and such placental alteration is crucial for the development of hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome. We aimed to analyze for the first time human placental expression of MAPK p38alpha in pregnancies complicated by HELLP. The placental expression of MAPK p38alpha was investigated by semiquantitative polymerase chain reaction using cDNA extracted from placental tissue of 15 pregnancies with HELLP syndrome and 15 gestational age-matched controls. Seven patients with HELLP also had intrauterine fetal growth restriction (IUGR). In placenta from pregnancy complicated by HELLP, the expression of MAPK p38alpha is significantly decreased compared to the group with normal pregnancy (p < 0.001), while no difference was found between the HELLP and HELLP with IUGR subpopulations. Our study shows for the first time that MAPK p38alpha is expressed in the human placenta. Pregnancies with placental dysfunction and hypertensive complications are characterized by a significantly decreased expression of MAPK p38alpha. Our observations suggest that p38 MAPK signaling may be essential in placental angiogenesis and functioning. PMID- 19565357 TI - Diagnosing brain death by CT perfusion and multislice CT angiography. AB - INTRODUCTION: Although the diagnosis of brain death (BD) is usually based on clinical criteria, in sedated patients, ancillary techniques are needed. This study was designed to assess the accuracy of cerebral multislice computed tomographic angiography (CTA) and CT perfusion (CTP) in diagnosing BD. METHODS: Prospective observational study in 27 BD patients. RESULTS: All patients were diagnosed as BD based on clinical and electroencephalogram findings. After BD diagnosis, CTP was performed followed by 64-detector multislice CTA from the aortic arch to the vertex. Images were reconstructed from 0.5 mm sections. In 24 patients, a lack of cerebral blood flow (CBF) was detected by CTP, and CTA revealed luminal narrowing of the internal carotid artery in the neck and absence of anterior and posterior intracranial circulation (sensitivity 89%). CTA detected CBF exclusively in extracranial portions of the internal carotid and vertebral arteries. Two patients with anoxic brain injury and decompressive craniectomy showed CBF in the CTA such that the CTP results were considered false negatives, given BD had been confirmed by clinical and EEG findings, along with evoked potentials. In one clinically BD patient, in whom an alpha rhythm was detected in the electroencephalogram, CBF was only observed in the intracranial internal carotid with no posterior circulation noted. This patient was therefore considered exclusively brain stem dead. CONCLUSIONS: The radiological protocol used shows a high sensitivity and excellent specificity for detecting the cerebral circulatory arrest that accompanies BD. As a rapid, non-invasive, and widely available technique it is a promising alternative to conventional 4-vessel angiography. PMID- 19565358 TI - Central pontine myelinolysis in a patient with hyperosmolar hyperglycemia and consistently normal serum sodium. AB - INTRODUCTION: Central pontine myelinolysis (CPM) is almost always described in association with a disturbance in sodium homeostasis, most commonly rapid correction of chronic hyponatremia. It has only rarely been described in patients with disturbances of serum osmolality in the absence of abnormal serum sodium concentrations. METHODS: Case report. RESULTS: A 93 year-old-man developed marked gait ataxia 2 days after the diagnosis and treatment of hyperosmolar hyperglycemia. MRI demonstrated a symmetric lesion in the central pons consisting of increased T2 signal intensity and restricted diffusion, consistent with CPM. Calculated serum osmolality on admission was 344 mOsm/kg and fell to 300 mOsm/Kg over 20 h. Serum sodium concentration stayed between 137 and 140 mEq/l throughout the admission. One month after admission, his ataxia had nearly completely resolved and the MRI changes had improved. CONCLUSION: CPM can develop in the setting of hyperosmolar hyperglycemia without abnormalities of sodium homeostasis. This supports the theory that the pathogenesis of CPM is dependent on a relatively hypertonic insult, which may occur independently of sodium abnormalities. CPM can present as isolated gait ataxia. Clinical manifestations of the disorder may show significant improvement despite a dramatic initial presentation. PMID- 19565359 TI - Plateau waves in head injured patients requiring neurocritical care. AB - OBJECT: Plateau waves often develop in neurointensive care patients. They are sudden increases in intracranial pressure (ICP) that lead to dramatic decreases of cerebral perfusion pressure (CPP) and can therefore contribute to ischemic secondary brain insult. The aim of this study was to analyze the occurrence of plateau waves in head injured patients requiring neurocritical care, their relation with cerebral autoregulation and impact on outcome. METHODS: Data were analyzed retrospectively in 444 head injured patients admitted to Neuroscience Critical Care Unit of Addenbrooke's Hospital in Cambridge, UK. Arterial blood pressure (ABP), intracranial pressure (ICP), heart rate (HR) were digitally recorded and derived indices calculated. Primary monitoring data, autoregulation indices, outcome of patients, initial CT findings (in a subgroup of patients), brain tissue monitoring data (in a subgroup) were compared between patients who developed plateau waves and those who did not. RESULTS: Plateau waves were observed in 109/444 patients (24.5%). They were significantly more frequent in younger patients. Impaired cerebrovascular pressure reactivity and depleted compensatory reserve were associated with vasodilatation on the top of the wave. Plateau waves were not associated with poorer outcome unless the episodes lasted for a long time (longer than 30-40 min). Plateau waves were more frequently seen in patients with lesser midline shift, lower volume of contusion on CT scan, absence of skull fractures, and lower brain tissue concentration of carbon dioxide. CONCLUSIONS: Plateau waves are frequent phenomenon. They are not associated with worse outcome unless they lead to sustained intracranial hypertension. PMID- 19565360 TI - Immunogenicity of Anti-TNF-alpha agents in autoimmune diseases. AB - Prognosis of several autoimmune diseases, especially rheumatoid arthritis (RA), ankylosing spondylitis, Crohn's disease (CD), and psoriasis, usually refractory to conventional treatment improved considerably with the introduction of tumor necrosis factor alpha (TNF-alpha) antagonistic agents, which is now available (infliximab, etanercept, and adalimumab). However, a portion of patients persists with active disease, infusion reactions, and relapses even during current biological therapy. One of the reasons for this is the associated immunogenicity to these drugs. The incentive for induction of antibodies against anti-TNF-alpha agent depends mainly on its constitution. Chimerical drugs have a higher capacity of inducing immunogenicity compared to completely human drugs. Among the three anti-TNF-alpha agents, this phenomenon has been studied mainly in patients using infliximab, especially in RA and CD. The prevalence of anti-infliximab antibodies in RA varies from 12% to 44% and seems to be inversely proportional to the level of seric infliximab and therapeutic response. The use of etanercept was associated to the development of anti-etanercept antibodies in 0% to 18% of patients, without apparent effect on effectiveness or adverse events. Studies with RA and CD patients show prevalence of anti-adalimumab antibodies from 1% to 87%. Immunosuppressive drug addiction can reduce the induction of anti-TNF-alpha antibodies. PMID- 19565361 TI - Neonatal estrogenic effects upon the male rat pituitary: early gonadotrophin attenuation precedes long-term recovery. AB - Neonatal exposure to potent estrogenic compounds can affect multiple components of the male reproductive system causing impaired development of the epithelium and overgrowth of stromal tissue in the epididymis, vas deferens, seminal vesicles, and prostate. However, very little is known about the direct effects of estrogenic compounds on the anterior pituitary gland. In this study we have investigated the effects of neonatal estrogenic exposure upon the anterior pituitary. Both the early- and late-stage effects of exposure to a synthetic estrogenic agent, diethylstilbestrol (DES), upon pituitary gonadotroph cell function were assessed. We administered either a high dose (10 microg) or a low dose (0.1 microg) of DES to male rats during their neonatal period (P2-12). Gonadotroph function, cell number and morphology shortly after DES treatment (P18) and during adulthood (P90) were assessed. At P18 there was a significant decrease in follicle stimulating hormone (FSH) immunoreactivity in the pituitary gonadotroph cells in the high DES dose treated rats compared to control animals. No significant change in luteinizing hormone (LH) was observed at either DES dose. In adulthood (P90), there was no significant difference in FSH or LH gonadotroph immunoreactivity between control rats and any dose of DES-treated rats. Therefore, despite acute and selective ablation of FSH expression the gonadotrophs were able to recover in adulthood, suggesting that perinatal estrogenic exposure was only temporarily deleterious. PMID- 19565363 TI - Primary gastrointestinal non-Hodgkin's lymphoma: clinicopathological and prognostic analysis. AB - The objective is to investigate the association between pathological type and clinical features, response to treatment and prognosis of primary gastrointestinal Non-Hodgkin's lymphoma (PGINHL). The clinicopathologic features, response to treatment and survival of 124 patients with PGINHL, were investigated retrospectively. Ninety-one patients were treated with surgery, most of which included combined therapy, while 32 patients received chemotherapy alone. Survival analysis was performed by Kaplan-Meier and Cox regression method. The most common immunophenotype was B-cell subtype. In 115 (92.7%) patients of B-cell lymphoma, mucosa-associated lymphoid tissue lymphoma (MALToma) and diffuse large B-cell lymphoma (DLBCL) were 55 and 50 patients, respectively. The patients of two pathological types had different clinical features including stage, B symptoms, sites of tumor, distant involvement, International Prognosis Index Score, size of tumor, and response to treatment. Both overall survival curve and multiple Cox regression analysis indicated that pathological type was statistically significant. The pathological subtype of PGINHL was an important prognostic factor. The patients with MALToma appear to have more favorable prognosis than those with DLBCL. PMID- 19565364 TI - Acute renal failure related to oxaliplatin-induced intravascular hemolysis. AB - Oxaliplatin is an effective chemotherapeutic agent frequently used in the treatment of colorectal carcinoma. Rare cases of renal failure and hemolytic reactions have been reported as separate side effects of oxaliplatin. Here we present a clinical picture of immune-related intravascular hemolysis and acute tubular necrosis in a patient receiving this drug. This case suggests a mechanistic explanation of renal failure in patients treated with oxaliplatin. PMID- 19565365 TI - Surgical versus medical treatment for secondary post-partum hemorrhage. AB - BACKGROUND: Secondary post-partum hemorrhage (PPH) is defined as any abnormal bleeding from the birth canal occurring between 24 hours and 12 weeks postnatally. Treatment usually falls into one of the two categories: surgical evacuation of the uterus or medical treatment. OBJECTIVE: To compare the two different clinical approaches and the implications on future fertility. STUDY DESIGN: A retrospective study. SETTING: From 1990 to 2002, 168 women diagnosed with late PPH were admitted to the Hadassah Medical Centers in Jerusalem. The cases were divided into two groups according to the planned initial treatment: primary surgical treatment vs. primary medical treatment. RESULTS: Primary surgical treatment was associated with significantly more primary negative events (p=0.01). After the primary event, primary surgical treatment was associated with fewer future deliveries (p=0.04) and resulted in increased rate of secondary infertility of borderline significance (p=0.06). CONCLUSIONS: Our results show that secondary PPH is related to high rates of immediate and long-term complications. It is possible that a conservative medical approach for secondary PPH may be superior to surgical treatment. PMID- 19565362 TI - On and around microtubules: an overview. AB - Microtubules are hollow tubes some 25 nm in diameter participating in the eukaryotic cytoskeleton. They are built from alphabeta-tubulin heterodimers that associate to form protofilaments running lengthwise along the microtubule wall with the beta-tubulin subunit facing the microtubule plus end conferring a structural polarity. The alpha- and beta-tubulins are highly conserved. A third member of the tubulin family, gamma-tubulin, plays a role in microtubule nucleation and assembly. Other members of the tubulin family appear to be involved in microtubule nucleation. Microtubule assembly is accompanied by hydrolysis of GTP associated with beta-tubulin so that microtubules consist principally of 'GDP-tubulin' stabilized at the plus end by a short 'cap'. An important property of microtubules is dynamic instability characterized by growth randomly interrupted by pauses and shrinkage. Many proteins interact with microtubules within the cell and are involved in essential functions such as microtubule growth, stabilization, destabilization, and interactions with chromosomes during cell division. The motor proteins kinesin and dynein use microtubules as pathways for transport and are also involved in cell division. Crystallography and electron microscopy are providing a structural basis for understanding the interactions of microtubules with antimitotic drugs, with motor proteins and with plus end tracking proteins. PMID- 19565366 TI - Which type of mid-urethral sling procedure should be chosen for treatment of stress urinary incontinance with intrinsic sphincter deficiency? Tension-free vaginal tape or transobturator tape. AB - OBJECTIVE: To compare tension-free vaginal tape (TVT) and transobturator tape (TOT) for surgical treatment of stress urinary incontinence (SUI) with intrinsic sphincter deficiency. DESIGN: Retrospective study. SETTING: Gynecology department, Bakirkoy Women and Childrens' Hospital, Istanbul. SAMPLE: Three hundred women urodynamically diagnosed with stress incontinence with intrinsic sphincter deficiency underwent synthetic mid-urethral sling procedures (TVT = 180, TOT = 120). METHODS: Before the operation, a complete medical history was taken and a gynecologic examination was performed. Subjects with detrusor overactivity or previous sling surgery were excluded. Clinical checkups were conducted at 3, 6, and 12 months, and then annually. MAIN OUTCOME MEASURES: Intraoperative complications, postoperative complications, and subjective cure rates. RESULTS: There were no significant differences in demographics between the TVT and TOT groups: mean age, parity, body mass index, menopausal status, and hormone replacement therapy. At a mean follow-up of 31.2 +/- 9.1 (range 12-46) months, the overall cure rates were 78.3% for TVT and 52.5% TOT (p < 0.0001). The risk of treatment failure in women who received TOT was 4.9 times higher than in women who underwent TVT. There were no significant differences in perioperative and postoperative complication rates between the two groups. CONCLUSION: TVT appears to be the preferable surgical option for the treatment of SUI with intrinsic sphincter deficiency. PMID- 19565367 TI - Surface roughness and gloss of indirect composites etched with acidulated phosphate fluoride solution. AB - OBJECTIVE: To evaluate the surface characteristics of indirect composites etched with acidulated phosphate fluoride (APF) solution. MATERIAL AND METHODS. APF solution was applied to specimens of six composite materials. The surface morphological change was measured with a confocal laser scanning microscope. Gloss measurement and scanning electron microscope (SEM) observation were also performed. RESULTS: The APF-treated surface showed various patterns due to the difference in the filler incorporated in each composite. The surface properties could be evaluated precisely from the surface roughness parameters. The arithmetical mean deviation of roughness profile (Ra) parameter of the specimens treated with APF solution was higher than those of the specimens untreated with APF, except New Meta Color Infis (IF). The difference in etching influence of each composite material was shown conclusively in the maximum height of the profile (Rz) parameter. Gradia (GR) and Gradia Forte (GF) were etched three times more deeply than that of IF categorized microfilled composite. The mean width of the profile element (RSm) decreased significantly after APF treatment, except in IF. Gloss was reduced apparently in all materials, indicating that gloss reduction was sensitive to slight surface changes. CONCLUSIONS: Specific filler particles of prosthodontic composite materials were etched by APF application. The surface roughness parameters, such as Rz and RSm, properly described various surface topographic features. Gloss was strongly correlated to surface roughness, as defined by these parameters, and especially to the initial change of surface roughness. PMID- 19565368 TI - Influence of two different fluoride compounds and an in vitro pellicle on the amount of KOH-soluble fluoride and its retention after toothbrushing. AB - OBJECTIVE: To determine the influence of two different fluoride compounds and an in vitro pellicle on KOH-soluble fluoride formation - its retention and resistance to toothbrushing. MATERIAL AND METHODS: Forty bovine incisors were randomly assigned to four groups (A-D). Of five samples prepared per tooth, one remained untreated and served as a baseline control. Groups A and B were pretreated with artificial saliva and groups C and D with human saliva. Groups A and C were treated with amine fluoride and groups B and D with sodium fluoride. After treatment, samples were brushed with 25, 50, and 75 brushing strokes. The amount of KOH-soluble fluoride formed on the enamel samples was measured at baseline, after application, and after 25, 50, and 75 brushing strokes. Fluoride uptake was calculated by unpaired t-tests and fluoride retention by paired t tests. RESULTS: No statistically significant differences in the KOH-soluble fluoride uptake of the groups that were pretreated (A vs B and C vs D) or treated equally (A vs C and B vs D) were observed. Retention of the KOH-soluble fluoride in the brushed samples was higher when the samples were pretreated with human saliva and treated with sodium fluoride than when the samples were pretreated with artificial saliva and treated with sodium fluoride. CONCLUSIONS: The fluoride compound and the acquired human in vitro pellicle have no influence on the uptake of KOH-soluble fluoride, but show a significant influence on abrasion resistance. PMID- 19565369 TI - Canal wall-down mastoidectomy with mastoid obliteration for pediatric cholesteatoma. AB - CONCLUSIONS: As cholesteatoma is widespread and erosive, it should be cleared away thoroughly, to keep a dry ear and conserve maximum hearing ability in children. Canal wall-down mastoidectomy with mastoid and epitympanum obliteration is a good choice for treatment of cholesteatoma in children. OBJECTIVE: To investigate canal wall-down mastoidectomy with mastoid and epitympanum obliteration for the treatment of cholesteatoma in children. PATIENTS AND METHODS: A retrospective analysis of all cases of pediatric middle ear cholesteatoma in children aged 5-12 years between 1999 and 2006 was conducted in Anhui Provincial Hospital. The follow-up information was completed. Forty-five patients (48 ears) were treated with canal wall-down mastoidectomy with mastoid and epitympanum obliteration and followed up for 2-5 years (mean 3.1 years). RESULTS: The recurrence rate and residual rate were 4.16% and 0%, respectively. All ears were dry within 8-10 weeks. Aural discharge was detected in two cases and controlled with antibiotic ear drops without complications. The dry ear rate was 95.8%. The air-bone gap closure had a mean of 17.2 +/- 2 dB. No obvious complication occurred after operations. PMID- 19565370 TI - Risk of cardiovascular outcomes in users of estradiol/dydrogesterone or other HRT preparations. AB - BACKGROUND: Use of postmenopausal hormone replacement therapy (HRT) used to be promoted to reduce the risk of ischemic cardiovascular diseases, a concept which has been challenged by results of the large Women's Health Initiative trial in users of estrogen and progestin. It is postulated that the type of progestin used in HRT affects the cardiovascular risk. METHODS: We used the UK-based General Practice Research Database to conduct a follow-up study with a nested case control analysis. We assessed and compared the risk of developing myocardial infarction, thrombotic stroke or venous thromboembolism in estradiol/dydrogesterone users, users of other HRT, or non-users of HRT. RESULTS: The incidence rates of myocardial infarction, thrombotic stroke and venous thromboembolism in estradiol/dydrogesterone users were 0.40 (95% confidence interval (CI) 0.18-0.76), 0.27 (95% CI 0.10-0.58) and 0.31 (95% CI 0.13-0.64) per 1000 person-years, respectively. As compared to non-users of HRT, the adjusted relative risk estimates (odds ratios) in the nested case-control analysis for estradiol/dydrogesterone users or users of other HRT were 1.06 (95% CI 0.48-2.36) and 1.12 (95% CI 0.84-1.51) for myocardial infarction, 0.50 (95% CI 0.21-1.22) and 1.18 (95% CI 0.94-1.48) for thrombotic stroke, and 0.84 (95% CI 0.37-1.92) and 1.42 (95% CI 1.10-1.82) for venous thromboembolism, respectively. CONCLUSION: The study provides evidence that estradiol/dydrogesterone use of several months to a few years is not associated with a higher risk of cardiovascular events than use of other HRT. PMID- 19565371 TI - Mesenchymal stromal cells promote or suppress the proliferation of T lymphocytes from cord blood and peripheral blood: the importance of low cell ratio and role of interleukin-6. AB - BACKGROUND AIMS: Mesenchymal stromal cells (MSC) have been shown to possess immunomodulatory functions and proposed as a tool for managing or preventing graft-versus-host disease (GvHD) as well as promoting clinical transplantation tolerance. We investigated the capacity of human bone marrow (BM) MSC to modulate the proliferation of T cells obtained from peripheral blood (PB) and umbilical cord blood (CB). We addressed the importance of the MSC:T-cell ratio, requirement for cell contact and impact of soluble factors on the MSC-mediated effects. We also analyzed whether regulatory T cells could be modulated by MSC in co cultures. METHODS: The effect of different MSC concentrations on T-cell proliferation induced by allogeneic, mitogenic or CD3/CD28 stimulation was analyzed using bromodeoxyuridine (BrdU) incorporation and carboxyfluorescein diacetate-succinimidyl ester (CFDA-SE) labeling. The level of regulatory T cells was assessed using quantitative real-time polymerase chain reaction (PCR) and flow cytometry analysis. RESULTS: MSC induced a dose- and contact-dependent inhibition of T-cell proliferation but lymphocytes from CB and PB were differentially affected. At low concentrations, MSC supported both CB and PB T cell proliferation, rather than inhibiting their proliferation. This supportive effect was contact independent and soluble factors such interleukin-6 (IL-6) appeared to be involved. Interestingly, among the expanded T-cell population in both CB and PB, regulatory T cells were increased and were a part of the new cells promoted by MSC at low doses. CONCLUSIONS: MSC represent an attractive tool for reducing the lymphocyte response by inhibiting T-cell activation and proliferation as well as promoting tolerance by maintaining and promoting the expansion of regulatory cells. Nevertheless, the dual ability of MSC to either sustain or suppress T-cell proliferation according to conditions should be considered in the context of clinical applications. PMID- 19565372 TI - Trans-tibial prosthesization in large area of residual limb wound: is it possible? A case report. PMID- 19565373 TI - Comfort and its measurement--a literature review. AB - PURPOSE: This review aims to identify measures suitable for evaluation of physical comfort in a range of clinical settings or specialised areas. METHOD: A review of literature included articles that evaluated an intervention, position, equipment or surface for comfort or discomfort. Electronic databases, hand searches and internet sources were used. RESULTS: In addition to several theoretical papers on comfort, 29 studies were identified that used 'comfort' or 'discomfort' as outcomes. There was a lack of consistency in measurement of comfort and researchers used a wide range of different scales and tools. Objective and subjective measures are described. The impact of symptoms, environmental variables and emotional factors was generally not considered. Two instruments evaluating subjective comfort or discomfort were identified that have been carefully developed and psychometrically tested. Both have potential for use in clinical practice and research - one developed for wheelchair seating and one for wearable computers. Suggestions for clinical evaluation of comfort based on the research and theoretical papers regarding measurement scales are made. CONCLUSION: Further work on development and validation of comfort assessment tools for other applications is needed. PMID- 19565374 TI - Satisfaction related to wheelchair use in older adults in both nursing homes and community dwelling. AB - PURPOSE: Older adults commonly use wheelchairs for mobility impairments regardless of their living situations. However, limited outcomes data are available to determine quality of the wheelchairs that older Americans are receiving, as well as their satisfaction with wheelchair service delivery programs. The purpose of this article was to analyze satisfaction data collected from three cohorts of older individuals living at nursing homes and in community settings. Ninety participants were residents in VA (n = 60). METHODS: One hundred thirty two older adults completed the standardized Quebec User Evaluation of Satisfaction with Assistive Technology. Ninety participants were residents in VA (n = 30) or private nursing homes (n = 30), and 42 were community dwelling participants. Those enrolled were either independent manual or power wheelchairs users. RESULTS: The community dwelling group reported significantly higher levels of satisfaction with their manual wheelchairs and service delivery when compared with the private nursing home participants (4.3 versus 2.9, p = 0.002). The satisfaction was also higher for VA nursing homes than private (3.9 versus 2.9, p = 0.004). No significant differences were reported for satisfaction levels regarding powered wheelchairs use between three groups. CONCLUSION: Level of satisfaction should be incorporated as an outcomes measure for evaluating wheelchair prescriptions and service delivery programs. This study also supports the establishment of a re-evaluation process of wheelchair fit as the users' age, to aide in provision of the best quality wheelchairs and service delivery programs. PMID- 19565375 TI - Impact of wheelchair acquisition on social participation. AB - PURPOSE: Efficient mobility could be a prerequisite to carrying out many daily activities and social roles (social participation). The aim of this study was to assess the impact of wheelchair acquisition on social participation. METHODS: Single group pre/post design where the intervention was the acquisition of a wheelchair paid for by the provincial government. Data were collected retrospectively from the participants' medical files. Individuals were excluded if they received an assistive device other than a wheelchair or contacted the centre only for wheelchair repairs. Social participation was measured using the Reintegration to Normal Living Index (RNLI) questionnaire. RESULTS: The sample (n = 42) had a mean age of 64.2 +/- 18.5 years, and 50% of them (n = 21) did not have a wheelchair before the intervention. The total RNLI scores pre- (46.9/100 +/- 24.7) and post-acquisition (29.7/100 +/- 18.5) showed a significant improvement in participation (p < 0.001). No difference was found between those who had their first wheelchair (n = 21) compared with replacement. Single-item analysis of the RNLI showed a significant difference for 5 of the 11 items. Age and diagnosis were significantly correlated (p < 0.05) with some of the items. CONCLUSION: Social participation improved significantly following wheelchair acquisition although confounding variables may have contributed to this improvement. PMID- 19565377 TI - Two persons with severe post-coma motor impairment and minimally conscious state use assistive technology to access stimulus events and social contact. PMID- 19565379 TI - A simple and useful hand orthosis for patients with amyotrophic lateral sclerosis: a simple web spacer for thumb opposition weakness. AB - PURPOSE: To evaluate the clinical efficacy of simple web spacer for thumb opposition weakness in patients with amyotrophic lateral sclerosis (ALS). METHOD: Observational survey. Setting. Hospital in Japan. RESULTS: The average grip strength with and without the web spacer was 8.25 +/- 1.74 kg and 8.13 +/- 1.91 kg, respectively. The lateral pinch strength was 0.83 +/- 0.45 kg and 0.93 +/- 0.47 kg. The palp pinch strength was 0.37 +/- 0.24 kg and 0.22 +/- 0.16 kg. In observation of activities of daily living (ADL) the movements of fingers required for palp pinching such as grasping buttons, flipping the pages of a book, and picking up small things were more easily carried out when wearing. CONCLUSIONS: The simple web spacer, which improves precision hand motion and facilitates grasping, should be considered in the treatment of patients with ALS. PMID- 19565378 TI - Public computing options for individuals with cognitive impairments: survey outcomes. AB - PURPOSE: To examine availability and accessibility of public computing for individuals with cognitive impairment (CI) who reside in the USA. METHOD: A telephone survey was administered as a semi-structured interview to 145 informants representing seven types of public facilities across three geographically distinct regions using a snowball sampling technique. An Internet search of wireless (Wi-Fi) hotspots supplemented the survey. RESULTS: Survey results showed the availability of public computer terminals and Internet hotspots was greatest in the urban sample, followed by the mid-sized and rural cities. Across seven facility types surveyed, libraries had the highest percentage of access barriers, including complex queue procedures, login and password requirements, and limited technical support. University assistive technology centres and facilities with a restricted user policy, such as brain injury centres, had the lowest incidence of access barriers. CONCLUSION: Findings suggest optimal outcomes for people with CI will result from a careful match of technology and the user that takes into account potential barriers and opportunities to computing in an individual's preferred public environments. Trends in public computing, including the emergence of widespread Wi-Fi and limited access to terminals that permit auto-launch applications, should guide development of technology designed for use in public computing environments. PMID- 19565380 TI - The effect of shaped wheelchair cushion and lumbar supports on under-seat pressure, comfort, and pelvic rotation. AB - AIM: A wheelchair seat and position help clients perform daily activities. The comfort of the wheelchair can encourage clients to participate in daily activities and can help prevent future complications. PURPOSE: This study evaluates how a shaped seat-cushion and two different back supports affect under seat pressure, comfort, and pelvic rotation. METHOD: Thirty healthy subjects were tested using two differently equipped manual wheelchairs. One wheelchair had a Velcro adjustable back seat and a plane seat-cushion. The other wheelchair had a non-adjustable sling-back seat and a plane cushion. The second wheelchair was also equipped with a shaped cushion and/or a detachable lumbar support. Under seat pressure, estimated comfort, and pelvic rotation were measured after 10 min in each wheelchair outfit. RESULTS: Peak pressure increased with the shaped cushion compared to the plane cushion. No significant difference in estimated comfort was found. Pelvic posterior-rotation was reduced with the adjustable or detachable back-support irrespective of the shape of the seat cushion. CONCLUSIONS: To support a neutral pelvic position and spinal curvature, a combination of a shaped cushion and a marked lumbar support is most effective. PMID- 19565381 TI - Evaluation of a new device to prevent falls in persons with Parkinson's disease. AB - PURPOSE: To compare walking characteristics of individuals with Parkinson's disease (PD) using a new walking aid, the WalkAbout, with usual walking. METHOD: Fifteen subjects with PD were recruited. Subjects walked in their usual fashion and then walked again in the WalkAbout. Gait parameters, 5-min walk, and oxygen consumption were recorded. RESULTS: Stride lengths were shorter when using the WalkAbout. On an average, the distance walked in 5 min and the oxygen uptake was not different when walking with the WalkAbout compared with the usual walk. Eight subjects (responders) walked further with the WalkAbout compared to their usual walk (164.90 +/- 55.72 m vs. 140.82 +/- 55.94 m). Seven subjects (non-responders) walked a shorter distance while using the WalkAbout compared to their usual walk (241.79 +/- 73.06 m vs. 281.24 +/- 82.83 m). Compared to non-responders, responders were older, had more severe disability, and were more likely to use an assistive device for walking. Responders walked more slowly, had a shorter stride length, and walked shorter distances in 5 min than non-responders. CONCLUSION: The WalkAbout may help persons with PD who have more severe disability to walk farther. These data could be beneficial in selecting a helpful walking aid for persons with PD. PMID- 19565382 TI - Special issue on web accessibility research. PMID- 19565383 TI - From Web accessibility to Web adaptability. AB - PURPOSE: This article asserts that current approaches to enhance the accessibility of Web resources fail to provide a solid foundation for the development of a robust and future-proofed framework. In particular, they fail to take advantage of new technologies and technological practices. The article introduces a framework for Web adaptability, which encourages the development of Web-based services that can be resilient to the diversity of uses of such services, the target audience, available resources, technical innovations, organisational policies and relevant definitions of 'accessibility'. METHOD: The article refers to a series of author-focussed approaches to accessibility through which the authors and others have struggled to find ways to promote accessibility for people with disabilities. These approaches depend upon the resource author's determination of the anticipated users' needs and their provision. Through approaches labelled as 1.0, 2.0 and 3.0, the authors have widened their focus to account for contexts and individual differences in target audiences. Now, the authors want to recognise the role of users in determining their engagement with resources (including services). To distinguish this new approach, the term 'adaptability' has been used to replace 'accessibility'; new definitions of accessibility have been adopted, and the authors have reviewed their previous work to clarify how it is relevant to the new approach. RESULTS: Accessibility 1.0 is here characterised as a technical approach in which authors are told how to construct resources for a broadly defined audience. This is known as universal design. Accessibility 2.0 was introduced to point to the need to account for the context in which resources would be used, to help overcome inadequacies identified in the purely technical approach. Accessibility 3.0 moved the focus on users from a homogenised universal definition to recognition of the idiosyncratic needs and preferences of individuals and to cater for them. All of these approaches placed responsibility within the authoring/publishing domain without recognising the role the user might want to play, or the roles that other users in social networks, or even Web services might play. CONCLUSION: Adaptability shifts the emphasis and calls for greater freedom for the users to facilitate individual accessibility in the open Web environment. PMID- 19565385 TI - Experimental evaluation of usability and accessibility of heading elements. AB - PURPOSE: To evaluate if marking up heading elements of Web sites improves usability and accessibility. METHOD: Task completion times and questionnaire responses were collected from sighted and blind users for two kinds of Web sites: sites marked up appropriately with heading elements, and sites with the same visual appearance but with no heading elements marked up. The experiment was carried out with user agents that could navigate through heading elements. RESULTS: (1) Heading elements improved usability both for sighted and blind users in terms of significantly reduced task completion time and also higher user satisfaction with the structured site. (2) A significant difference in accessibility (i.e., if reduction of task completion time is larger for blind users than sighted users) was not observed when heading elements were marked up. The lack of significant effect in this case might have been due to blind users making use of the user-agent's navigation functions to find the target information efficiently when the heading elements were not present. CONCLUSIONS: A combination of content with heading elements marked up, user agents with functions that utilise structure markup, and users with sufficient knowledge about user agents functionality, improves usability for both sighted and blind users and may improve accessibility for blind users. PMID- 19565384 TI - Web accessibility and open source software. AB - A Web browser provides a uniform user interface to different types of information. Making this interface universally accessible and more interactive is a long-term goal still far from being achieved. Universally accessible browsers require novel interaction modalities and additional functionalities, for which existing browsers tend to provide only partial solutions. Although functionality for Web accessibility can be found as open source and free software components, their reuse and integration is complex because they were developed in diverse implementation environments, following standards and conventions incompatible with the Web. To address these problems, we have started several activities that aim at exploiting the potential of open-source software for Web accessibility. The first of these activities is the development of Adaptable Multi-Interface COmmunicator (AMICO):WEB, an infrastructure that facilitates efficient reuse and integration of open source software components into the Web environment. The main contribution of AMICO:WEB is in enabling the syntactic and semantic interoperability between Web extension mechanisms and a variety of integration mechanisms used by open source and free software components. Its design is based on our experiences in solving practical problems where we have used open source components to improve accessibility of rich media Web applications. The second of our activities involves improving education, where we have used our platform to teach students how to build advanced accessibility solutions from diverse open source software. We are also partially involved in the recently started Eclipse projects called Accessibility Tools Framework (ACTF), the aim of which is development of extensible infrastructure, upon which developers can build a variety of utilities that help to evaluate and enhance the accessibility of applications and content for people with disabilities. In this article we briefly report on these activities. PMID- 19565386 TI - Tool independence for the Web Accessibility Quantitative Metric. AB - The Web Accessibility Quantitative Metric (WAQM) aims at accurately measuring the accessibility of web pages. One of the main features of WAQM among others is that it is evaluation tool independent for ranking and accessibility monitoring scenarios. This article proposes a method to attain evaluation tool independence for all foreseeable scenarios. After demonstrating that homepages have a more similar error profile than any other web page in a given web site, 15 homepages were measured with 10,000 different values of WAQM parameters using EvalAccess and LIFT, two automatic evaluation tools for accessibility. A similar procedure was followed with random pages and with several test files obtaining several tuples that minimise the difference between both tools. One thousand four hundred forty-nine web pages from 15 web sites were measured with these tuples and those values that minimised the difference between the tools were selected. Once the WAQM was tuned, the accessibility of 15 web sites was measured with two metrics for web sites, concluding that even if similar values can be produced, obtaining the same scores is undesirable since evaluation tools behave in a different way. PMID- 19565387 TI - Editing Wikipedia content by screen reader: easier interaction with the Accessible Rich Internet Applications suite. AB - PURPOSE: This study aims to improve Wikipedia usability for the blind and promote the application of standards relating to Web accessibility and usability. METHOD: First, accessibility and usability of Wikipedia home, search result and edit pages are analysed using the JAWS screen reader; next, suggestions for improving interaction are proposed and a new Wikipedia editing interface built. Most of the improvements were obtained using the Accessible Rich Internet Applications (WAI ARIA) suite, developed by the World Wide Web Consortium (W3C) within the framework of the Web Accessibility Initiative (WAI). Last, a scenario of use compares interaction of blind people with the original and the modified interfaces. RESULTS: Our study highlights that although all contents are accessible via screen reader, usability issues exist due to the user's difficulties when interacting with the interface. The scenario of use shows how building an editing interface with the W3C WAI-ARIA suite eliminates many obstacles that can prevent blind users from actively contributing to Wikipedia. CONCLUSION: The modified Wikipedia editing page is simpler to use via a screen reader than the original one because ARIA ensures a page overview, rapid navigation, and total control of what is happening in the interface. PMID- 19565388 TI - WAI-ARIA live regions and channels: ReefChat as a case example. AB - Web 2.0, enabled by the AJAX architecture, has given rise to new levels of user interaction with web pages. Many of these new and extremely popular Web 2.0 pages are better classified as fully-fledged applications, some examples being Google Maps, Google Docs and Flickr. Unfortunately, accessibility support in most AJAX applications is lacking. The Web Accessibility Initiative-Accessible Rich Internet Applications markup presents a solution to making these applications accessible. This article will look at current ARIA documentation available for Web 2.0 developers, a new innovation involving channels, and present a real life example of an ARIA based solution involving an accessible AJAX chat, ReefChat, and an ARIA compatible screen reader, Fire Vox. PMID- 19565389 TI - Enabling web users and developers to script accessibility with Accessmonkey. AB - Efficient web access remains elusive for blind computer users. Previous efforts to improve web accessibility have focused on developer awareness, automated improvement, and legislation, but these approaches have left remaining concerns. First, while many tools can help produce accessible content, most are difficult to integrate into existing developer workflows and rarely offer specific suggestions that developers can implement. Second, tools that automatically improve web content for users generally solve specific problems and are difficult to combine and use on a diversity of existing assistive technology. Finally, although blind web users have proven adept at overcoming the shortcomings of the web and existing tools, they have been only marginally involved in improving the accessibility of their own web experience. In a step toward addressing these concerns, we have developed Accessmonkey, a common scripting framework that web users, web developers and web researchers can use to collaboratively improve accessibility. This framework advances the idea that Javascript and dynamic web content can be used to improve inaccessible content instead of being a cause of it. Using Accessmonkey, web users and developers on different platforms and with potentially different goals can collaboratively make the web more accessible. In this article, we first present the design of the Accessmonkey framework and offer several example scripts that demonstrate the utility of our approach. We conclude by discussing possible future extensions that will provide easy access to scripts as users browse the web and enable non-technical blind users to independently create and share improvements. PMID- 19565392 TI - Effectiveness of Lee Silverman Voice Treatment (LSVT) on hypernasality in non progressive dysarthria: the need for further research. AB - BACKGROUND: Hypernasality is a common feature of non-progressive dysarthria. However, limited research has investigated the effectiveness of treatments for this impairment. Preliminary research has revealed positive effects on nasalance when using increased loudness in certain non-progressive dysarthric speakers. However, the long-term effects of loud speech on nasalance as part of a structured intervention such as Lee Silverman Voice Treatment (LSVT) are yet to be investigated in this population. AIMS: The study aimed to investigate the short- and long-term effects of LSVT on hypernasality (perceptual ratings and degree of nasalance) in non-progressive dysarthria; and secondly, to evaluate the effects of traditional dysarthria therapy on these same measures, in comparison with the effects of LSVT. METHODS & PROCEDURES: Ten non-progressive dysarthric speakers with varying levels of hypernasality (taken from a larger research study) were randomly allocated to receive LSVT((R)) (n = 5) or individually tailored traditional dysarthria therapy (n = 5). Both treatments were administered four times weekly for 4 weeks (that is, 16 x 1-hour sessions). Participants were assessed twice before treatment, twice immediately post treatment, and twice at follow-up 6 months post-treatment using a perceptual rating task performed by two independent speech pathologists, and the Nasometer. Changes to individual mean nasalance scores were compared against clinically significant criterion and perceptual ratings were analysed descriptively. OUTCOMES & RESULTS: Three out of five participants demonstrated reductions in perceived hypernasality immediately following LSVT, but these changes were maintained at follow-up for only one participant. Two of these three participants demonstrated a corresponding reduction in mean nasalance. Limited changes in perceived hypernasality and nasalance scores were found following traditional dysarthria therapy, with only one participant exhibiting reduced nasalance at follow-up. CONCLUSIONS & IMPLICATIONS: Due to the small sample size in the present research and variability between participants, further exploration into the effects of LSVT on nasality with a larger population with different dysarthria types is essential. PMID- 19565394 TI - The voice handicap index with post-laryngectomy male voices. AB - BACKGROUND: Surgical treatment for advanced laryngeal cancer involves complete removal of the larynx ('laryngectomy') and initial total loss of voice. Post laryngectomy rehabilitation involves implementation of different means of 'voicing' for these patients wherever possible. There is little information about laryngectomees' perception of their changed voice quality and communication status. Surgical voice restoration (SVR) has become the 'gold standard' rehabilitation, but there continue to be patients who use other methods of communication. There is no clear evidence comparing patients' perception of their voice handicap across different types of alaryngeal communication. AIMS: To compare the self-assessed vocal handicap of laryngectomees using SVR with those using non-SVR methods of post-laryngectomy communication. METHODS & PROCEDURES: Potential participants were identified from one Head and Neck cancer centre in South Wales. They included both male and female participants using all methods of post-laryngectomy communication. Each patient's Voice Handicap Index (VHI) score, sub-set scores, and group means were calculated. Two major confounding factors: age and time since surgery, and communication method (SVR/non-SVR), were considered to identify factors, other than method of communication, which may influence rehabilitation outcomes. OUTCOMES & RESULTS: A total of 71 questionnaires were sent out and 62 (82%) were returned from 35 patients who had undergone SVR and 27 patients who used non-SVR methods of communication. Of the non-SVR group, twelve used oesophageal voice, eleven an electrolarynx, two writing and two mouthing for communication. The gender ratio (53:9), age (43-90 years) and time since surgery (1-40 years) were broadly representative of this population, but because of the small number of females, we excluded the women from further analysis. Individual VHI scores ranged from 4 to 106. Both the SVR and non-SVR group mean scores: 44.7 and 50.9, were within the range of moderately severe voice handicap. There was no significant difference between the groups for total VHI scores or two of the three sub-domains, nor any significant effect on voice handicap due to the confounding factors assessed: age or time since surgery. The total VHI score was better by 6.5 ( - 4.9 to 17.9) points in the SVR group (p = 0.3), probably reflecting the literature reporting superior voice in SVR. CONCLUSIONS & IMPLICATIONS: The data suggest that where patient-assessed quality of life is concerned, SVR and non-SVR outcomes are comparable. This is an important consideration when planning and carrying out treatment recommendations. The study has clear clinical implications; understanding the potential of all methods of post-laryngectomy communication is essential for holistic patient management. PMID- 19565397 TI - Treatment generalization and executive control processes: preliminary data from Chinese anomic individuals. AB - BACKGROUND: While various treatment approaches have been shown to be effective in remediating word-finding difficulties in aphasic individuals, interest has recently been directed at the role of executive functions in affecting treatment outcomes. AIMS: To examine the existence of a possible relationship between treatment generalization and executive control abilities. METHODS & PROCEDURES: An identical treatment protocol using the English alphabet as letter cues to facilitate name retrieval was applied to five Cantonese-speaking anomic individuals. A series of language and cognitive tasks were administered to help identify which deficit(s) would affect treatment results. OUTCOMES & RESULTS: All five participants demonstrated improvement in naming treated items. However, only two subjects with better inhibitory control exhibited significant gains in naming phonologically related untrained items. CONCLUSIONS & IMPLICATIONS: The preliminary results not only converge on current findings of the role of executive functions in rehabilitation results, but also identify in particular how inhibitory control may affect the outcomes of anomia therapy. PMID- 19565400 TI - Validation of dual-energy x-ray absorptiometry and foot-foot impedance against deuterium dilution measures of fatness in children. AB - OBJECTIVE: To determine the validity of estimation of body fatness by dual-energy x-ray absorptiometry (DXA) and foot-foot bio-electrical impedance (BIA). METHODS: In 176, 11-12-year-olds (84 boys; 92 girls) body fatness was measured using total body water (TBW), derived from deuterium oxide dilution space. Body fatness was also estimated from DXA and BIA. Methods were compared by regression and by Bland Altman analysis using TBW measures as the reference. RESULTS: In boys, mean fat mass from TBW was 9.8 kg (standard deviation, SD=6.1); bias by DXA estimated fat mass was +0.9 kg (limits of agreement -2.2 to +4.1) and bias for BIA was -5.2 kg (limits of agreement +0.5 to -10.8). In boys, regression analysis indicated significant differences in slope (p<0.001) for DXA, and both slope (p < 0.001) and intercept (p < 0.001) for BIA. In girls, mean fat mass from TBW was 12.1 kg (SD 7.7); bias for DXA was +1.2 kg (limits of agreement -1.9 to +5.1) and bias for BIA was -0.2 kg (limits of agreement -5.4 to +5.1). In girls, regression analysis indicated significant differences for slope and intercept (p<0.001 in all cases) for both DXA and BIA. CONCLUSIONS: Errors in estimation of fat mass using BIA and DXA can be very large, and the direction of error can differ between the sexes. PMID- 19565401 TI - Perinatal and childhood risk factors for overweight in a provincial sample of Canadian Grade 5 students. AB - BACKGROUND: The risk of obesity is determined by a complex interaction of prenatal, lifestyle, genetic, and environmental factors. OBJECTIVE: To investigate the differential impact of prenatal, child, and family factors on body weight status in childhood. METHODS: The current study links population based survey data of Grade 5 students who participated in the 2003 Children's Lifestyle and School Performance Study in Nova Scotia, Canada, with a provincial perinatal registry. Multilevel logistic regression was used to investigate the association between prenatal, lifestyle, and socioeconomic factors and childhood overweight. RESULTS: Of the 4 298 participating children, 3 426 (80%) could be linked with information in the perinatal registry. Small-for-gestational age infants appeared to be less likely to be overweight at age 10 or 11 years (odds ratio [OR]=0.68) whereas those born large-for-gestational age were more likely to be overweight (OR=1.23). Maternal pre-pregnancy weight was associated with childhood overweight (OR=4.42 for >80 kg vs. <60 kg) as were the child's physical activity (OR=0.65 for >7x/week vs. < or =2x/week) and screen time (OR=1.82 for >6 h/day vs. < or =1h/day). Smoking during pregnancy increased the child's odds for being overweight (OR=1.42 for >0.5 packs/day vs. none). Children living in neighborhoods with higher housing values were less likely to be overweight (OR=0.68 for highest vs. lowest tertile). CONCLUSIONS: Overweight young women should be advised on the importance of healthy eating, active living and maintaining a healthy weight in the pre-pregnancy years to reduce the risk of overweight in their offspring. PMID- 19565402 TI - Is obesity associated with oxidative stress in children? AB - OBJECTIVE: We evaluated the presence of oxidative stress in obese children without co-morbidities. METHODS: The study population included 68 children (30 girls, 38 boys), between 6 and 14 years of age. The levels of markers of oxidative damage (malondialdehyde [MDA], and plasma carbonyl groups [CG]) and measures of antioxidant defense, such as the enzyme glutathione peroxidase (GPx) and low molecular scavengers (erythrocyte-reduced glutathione [GSH], alpha tocopherol and beta-carotene) were determined. Children were categorized in groups by the standard deviation score of body mass index (SDS-BMI). Twenty children were non-obese (SDS-BMI< or =1.33), and the 48 obese children (SDS-BMI> or =2) were further divided into two groups: SDS-BMI> or =3 (22 children) and > or =2 SDS-BMI<3 (26 children). RESULTS: The levels of MDA and CG were significantly higher (p<0.05) in children with SDS-BMI> or =3. The GPx activity was increased, while the GSH concentration was lower in obese children compared with non-obese children (p<0.01). There were no differences in serum alpha tocopherol and beta-carotene levels between groups. MDA was the sole marker of oxidative damage that was positively correlated with SDS-BMI, (r=0.35, p=0.015), and negatively with high-density lipoprotein cholesterol (HDL-C) (r=- 0.32, p=0.027). GPx was inversely related to total cholesterol (r=- 0.34, p=0.019). In multiple regression analysis, we confirmed that SDS-BMI and HDL-C were determinants of MDA. CONCLUSIONS: Severe childhood obesity is associated with oxidative stress. Thus, providing foods with high antioxidant capacity in addition to a hypocaloric diet is crucial for the treatment of obese children. PMID- 19565403 TI - Oral motor dysfunction in children with adenotonsillar hypertrophy--effects of surgery. AB - Adenotonsillar hypertrophy is associated with a wide range of problems. The enlargement causes obstructive symptoms and affects different functions such as chewing, swallowing, articulation, and voice. The objective of this study was to assess oral motor function in children with adenotonsillar hypertrophy using Nordic Orofacial Test-Screening (NOT-S) before and 6 months after surgery consisting of adenoidectomy combined with total or partial tonsil removal. A total of 67 children were assigned to either tonsillectomy (n=33) or partial tonsillectomy, 'tonsillotomy' (n=34); 76 controls were assessed with NOT-S and divided into a younger and older age group to match pre- and post-operated children. Most children in the study groups had oral motor problems prior to surgery including snoring, open mouth position, drooling, masticatory, and swallowing problems. Post-surgery oral motor function was equal to controls. Improvement was independent of surgery method. PMID- 19565404 TI - The inability to produce soft voice (IPSV): a tool to detect vocal change in school-teachers. AB - PURPOSE: This study investigated whether clinicians could detect voice changes reported by teachers, from self-ratings teachers conducted of their inability to produce soft voice (IPSV). METHODS: Ten teachers wore a vocal dosimeter and completed daily IPSV ratings approximately every 2 hours for 14 days. Following the 2 weeks of dosimetry, two speech clinicians specialized in voice rated the teachers' IPSV from dosimeter recordings. Teacher and clinician ratings were compared for each participant. RESULTS: Although agreement between teacher and clinician ratings was not significant, descriptive analyses demonstrated an average difference score of 1.7 (SD 1.4) between teacher and clinician ratings. CONCLUSIONS: This study supports the potential usefulness of the IPSV as a simple tool to detect voice changes in oneself or others. PMID- 19565405 TI - Speaking silence: the social construction of silence in autobiographical and cultural narratives. AB - Voice and silence are socially constructed in conversational interactions between speakers and listeners that are influenced by canonical cultural narratives which define lives and selves. Arguing from feminist and sociocultural theories, I make a distinction between being silenced and being silent; when being silenced is contrasted with voice, it is conceptualised as imposed, and it signifies a loss of power and self. But silence can also be conceptualised as being silent, a shared understanding that need not be voiced. More specifically, culturally dominant narratives provide for shared understandings that can remain silent; deviations from the norm call for voice, and thus in this case silence is power and voice expresses loss of power. At both the cultural and the individual level, there are tensions between culturally dominant and prescriptive narratives and narratives of resistance and deviation, leading to an ongoing dialectic between voice and silence. I end with a discussion of why, ultimately, it matters what is voiced and what is silenced for memory, identity and well-being. PMID- 19565406 TI - Severely and persistently mentally ill - a changing group. Ten years after the 1995 Swedish mental health care reform. AB - AIM: A mental health care reform was implemented in Sweden in 1995. The target group of the reform was persons considered severely and persistently mentally ill (SMI). The aim of this article was to study changes in the target group 10 years after the reform, i.e. changes in the group of persons defined as SMI. METHODS: Inventories of persons considered SMI were made in a municipality of Sweden in 1995/96, 2000/2001 and 2006 based on the same definition. The invented persons were interviewed. The same interview procedures, questionnaires and need assessments were used on all three occasions. The results were based on comparing the new groups considered SMI in 2000/2001 and 2006. RESULTS: In 2006, the group of persons considered SMI was younger. Fewer persons with a diagnosis of schizophrenia and more persons with a diagnosis of mood disorders or addictive problems were included. Problems concerning relations and physical health were more evident. A greater part had all their income from social security in 2006, and fewer persons were occupied in some kind of salaried employment, full-time or half-time, regular or sheltered. CONCLUSION: The target group of the reform had changed and other groups of persons were considered SMI in 2006. These new groups require other strategies of care. PMID- 19565407 TI - Are specific guidelines necessary for treatment of esophageal varices in the Nordic countries? PMID- 19565408 TI - Mycobacterial infections in coal workers' pneumoconiosis patients in South Korea. AB - Coal workers' pneumoconiosis (CWP) is the most common occupational disease in South Korea and is an important factor in the development of infections with Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacteria (NTM). In the current study, we identified mycobacterial species that cause pulmonary infections in CWP patients, using rpoB DNA-PCR-restriction analysis. Among the 129 CWP patients studied, 35 (27.1%) were diagnosed as having mycobacterial infections. Among these, the proportion of NTM infections (21/35, 60.0%) was higher than that for MTB infections (14/35, 40.0%). Of the 21 NTM strains, the most common was M. intracellulare (6/21, 28.6%), followed by M. avium (5/21, 23.8%). We also compared the characteristics of CWP patients between the MTB and NTM infection groups. A higher proportion of CWP patients with NTM infections compared with those with MTB infections had a history of having been involved in rock work (38.1% vs 21.4%), and had complicated CWP (85.7% vs 35.7%) and a past history of TB treatment (61.9% vs 50.0%). We also discovered 3 MTB mutants that are resistant to first-line anti-TB drugs, in CWP patients. These results demonstrate the features of pulmonary mycobacterial infections with a predominance of NTM in CWP patients in South Korea. PMID- 19565409 TI - P-drive on-road: internal scale validity and reliability of an assessment of on road driving performance in people with neurological disorders. AB - The aim of this study was to determine internal scale validity and reliability of P-Drive on-road by observing driving performance among people with neurological disorders. The study comprised a consecutive sample of 205 persons with stroke, dementia, or mild cognitive impairment who were remitted for and performed a standardized test in real traffic. Their driving abilities were evaluated with a new assessment tool, P-Drive on-road. Aspects of validity and reliability of the tool were analyzed using a Rasch rating scale model. The results indicated that the rating scale functioned as intended and a principal component analysis indicated that the items formed a unidimensional scale. Acceptable levels of person response validity were found and the scale was also able to distinguish between the abilities of the drivers with a person separation reliability of 0.90. The results indicate that P-Drive on-road is valid and reliable for producing a linear measure of driving ability in people with stroke, dementia and mild cognitive impairment, and has the potential to become a clinically useful assessment tool for on-road driving tests. PMID- 19565410 TI - Consumer acoustic reflectometry by parents in detecting middle-ear fluid among children undergoing tympanostomy. AB - OBJECTIVE: This study aimed at evaluating the diagnostic accuracy of a consumer model acoustic reflectometer in the hands of parents in the detection of middle ear fluid among children. DESIGN: Prospective diagnostic study according to STARD guidelines. Acoustic reflectometry recorded by the parents was compared with otomicroscopic myringotomy with suction in general anaesthesia, performed immediately afterwards. SETTING: Secondary care (Kanta-Hame Central Hospital, Finland). PATIENTS: Pre-school (<7 years) children undergoing otomicroscopic myringotomy due to recurrent acute otitis media or suspicion of glue ear. MAIN OUTCOME MEASURES: Sensitivity, specificity, and positive and negative predictive values of acoustic reflectometry. RESULTS: Consumer model acoustic reflectometry performed by parents reached sensitivity of 94% (95% confidence interval 88-100%) and negative predictive value of 92% (83-100%) with result 1 (green colour) on a scale of 1-5. On the other hand, the method was not very specific and positive predictive value was poor. CONCLUSIONS: The negative screening result was very reliable. Thus, home screening of otitis among otitis-prone children and home follow-up of resolution of middle-ear fluid afterwards deserves to be studied in the home setting in the future. PMID- 19565411 TI - General practitioners trained in motivational interviewing can positively affect the attitude to behaviour change in people with type 2 diabetes. One year follow up of an RCT, ADDITION Denmark. AB - OBJECTIVE: To examine whether training GPs in motivational interviewing (MI) can improve type 2 diabetic patients' (1) understanding of diabetes, (2) beliefs regarding prevention and treatment, and (3) motivation for behaviour change. METHODS: A randomized controlled trial including 65 GPs and 265 type 2 diabetic patients. The GPs were randomized in two groups, one with and one without MI training. Both groups received training in target-driven intensive treatment of type 2 diabetic patients. The intervention was a 1(1/2)-day residential course in MI with (1/2)-day follow-up twice during the first year. The patient data stemmed from previously validated questionnaires. MAIN OUTCOME MEASURES: The Health Care Climates Questionnaire assesses the patient-doctor relationship and type of counselling. The Treatment Self-Regulation Questionnaire assesses the degree to which behaviour tends to be self-determined. The Diabetes Illness Representation Questionnaire assesses beliefs and understanding of type 2 diabetes. The Summary of Diabetes Self Care Activities assesses the extent of various self-care activities related to type 2 diabetes. RESULTS: The response rate to our questionnaires was 87%. Patients in the intervention group were significantly more autonomous and motivated in their inclination to change behaviour after one year compared with the patients from the control group. Patients in the intervention group were also significantly more conscious of the importance of controlling their diabetes, and had a significantly better understanding of the possibility of preventing complications. CONCLUSION: MI improved type 2 patients' understanding of diabetes, their beliefs regarding treatment aspects, their contemplation on and motivation for behaviour change. Whether our results can be sustained long term and are clinically relevant in terms of changes in risk profile advocates further research. PMID- 19565412 TI - Three- and four-digit ICD-10 is not a reliable classification system in primary care. AB - OBJECTIVE: The International Classification of Diseases 10th revision (ICD-10) is a standard international diagnostic classification for medical diagnoses. Reliable diagnostic coding is of high medical and epidemiological importance. Coding diagnoses with ICD-10 is the basis of reimbursement in some healthcare systems. DESIGN: The ICD-10 coding of each case was performed by two raters to investigate the inter-rater agreement. The degree of agreement was assessed using Cohen's kappa. Kappa was divided into two groups: Kappa > or = 0.61 meaning high or satisfactory and kappa < or = 0.6 (incl. < or = 0.000 and 0.000*) meaning low or unsatisfactory. SUBJECTS: Cross-sectional data were collected from 8877 randomly selected patients. The 209 participating general practitioners used a standardized data collection form. The first of the reasons for encounter was taken into account on new and chronic managed problems. RESULTS: Kappa values were satisfactory on the chapter level with on average 0.685 (chronic managed problems) and 0.675 (new managed problems). Kappa was unsatisfactory when the three-digit level was used (0.428) and lower for terminal codes (four-digit level) at 0.199 on average (chronic managed problems). For new managed problems the kappa values were at 0.384 (three-digit level) and 0.166 (four-digit level) on average. CONCLUSION: The ICD-10 is reliable for coding managed problems on the chapter level. Further refinement of ICD-10 with three- and four-digit codes leads to significant coding uncertainties. There is no reliable coding scheme that meets the demands of general practice. The use of coded data for healthcare reimbursement requires a simplification of ICD-10 to provide a realistic picture of morbidity. PMID- 19565413 TI - Making induced abortion safe and legal, worldwide. PMID- 19565414 TI - Increasing women's choices in medical abortion: a study of misoprostol 400 microg swallowed immediately or held sublingually following 200 mg mifepristone. AB - OBJECTIVES: To assess the efficacy and acceptability of two misoprostol regimens (400 microg oral or sublingual) following mifepristone for medical abortion. METHODS: Women seeking abortion with gestations of 56 days or less since onset of their last menstrual period were offered medical abortion as an alternative to a surgical procedure. A total of 207 eligible and consenting women were given mifepristone (200 mg oral) and the option of taking 400 microg misoprostol either orally or sublingually two days later, with the option of home-use. Two weeks later, treatment success, satisfaction, and the frequency and acceptability of side effects were assessed. RESULTS: Most women (97.6%) opted for home use of misoprostol and almost three quarters selected the oral route. Overall efficacy, acceptability of side effects and satisfaction were high in both groups. The success rate was lower after sublingual than after oral administration but not significantly so (91.3% vs. 96.3%, p = 0.23, RR: 0.93, 95% CI = 0.85-1.02). The frequency and average duration of side effects in both groups were comparable except for pain/cramps and fever/chills, which were more frequently associated with the sublingual route. CONCLUSIONS: This study re-emphasises the feasibility of integrating medical abortion into health services in Turkey and the potential to increase choices for women. PMID- 19565415 TI - Risk factors for induced abortions in St Petersburg, Estonia and Finland. Results from surveys among women of reproductive age. AB - OBJECTIVES: To compare the risk factors of self-reported abortion in St Petersburg, Estonia and Finland, which are neighbouring areas having different abortion rates. METHODS: Data from four population-based questionnaire surveys (Finland 1992 and 1999; St Petersburg 2003; Estonia 2004) were used. With the exception of the 1992 Finnish survey (based on interviews) all were postal surveys. There were 1070 respondents in Finland (78% and 52% response rates), 1147 (68%) in St Petersburg, and 5190 (54%) in Estonia. RESULTS: The prevalence of self-reported abortions was highest among women in St Petersburg and among the two oldest age groups of Russian-speaking Estonians. In all areas women with a history of abortion were more likely to have low education, children, a history of multiple partners and to have had their first intercourse when younger than 18 years. Insufficient use of reliable contraception, both at first and last intercourse, was the only risk factor that was more common in St Petersburg and in Estonia than in Finland. In Estonia, more Estonian than Russian-speaking women used reliable contraceptives. CONCLUSION: The comparison of three areas suggests that high abortion rates are related to low contraceptive use and not to other risky sexual behaviour. PMID- 19565416 TI - Profile of long-acting reversible contraception users in Europe. AB - OBJECTIVES: To assess the profile of long-acting reversible contraceptives (LARCs) users in Europe. METHODS: A random sample of women aged 15-49 years in 14 European countries (Germany, France, UK, Spain, Italy, Russian Federation, Estonia, Latvia, Lithuania, Austria, Czech Republic, Denmark, Norway, and Sweden) underwent web-based or computer-aided face-to-face interviews in June 2006. In this paper data pertaining to a subgroup of women using LARCs are presented. RESULTS: A total of 11,490 women participated in the full study. Of these, 1,188 (10%) women were LARC (hormonal implant, injectables, levonorgestrel-releasing intrauterine system [LNG-IUS], copper intrauterine device [Cu-IUD]) users. The age of the LARC users exceeded 30 years for 57-91% of them. Furthermore, more than half of them found convenience an extremely important factor when selecting the LARC as a contraceptive method. As compared to those wearing a Cu-IUD, women using hormonal LARCs experienced fewer physical and emotional symptoms that appeared or worsened during menstruation. CONCLUSIONS: LARCs have their place in the contraceptive market in Europe. The most popular LARCs among European women were the LNG-IUS and the Cu-IUD; both were mainly used by women who had children and had no wish to have more in the future. PMID- 19565417 TI - Teenagers and emergency contraception in the UK: a focus group study of salient beliefs using concepts from the Theory of Planned Behaviour. AB - OBJECTIVES: To explore teenagers' beliefs about emergency contraception (EC) within a Theory of Planned Behaviour (TPB) framework. METHODS: Six single sex focus groups comprising a total of 25 female and 23 male pupils aged 13-16 years conducted in schools in Central England. RESULTS: Attitudes to emergency contraception (EC) were mainly positive about the rewards of avoiding teenage pregnancy. Participants had positive beliefs about the effectiveness of EC, although knowledge of crucial time limits varied. EC use was more socially acceptable than teenage pregnancy, yet both outcomes were perceived negatively. Motivation to comply with social pressure was influenced by the appraisal of individuals' intentions. Participants reported high self efficacy in accessing EC, but had concerns over confidentiality and access. CONCLUSIONS: Desire to avoid pregnancy was high in this group, but practical factors and attitudes may be more important for those ambivalent about pregnancy. Adolescents perceive accessing EC as difficult, are concerned about confidentiality, and anticipate negative reactions from staff. Data support the TPB as a suitable framework for understanding attitudes to EC use. Further research should apply quantitative TPB measures to EC use in a wider teenage sample in order to identify potential psychological factors to target in an intervention. PMID- 19565418 TI - Oral contraception usage in relation to bone mineral density and bone turnover in adolescent girls. AB - OBJECTIVES: To compare the effect of a low-dose oral contraceptive (OC) containing 30 microg ethinyloestradiol (EE) with that of an ultra-low-dose OC containing 15 microg EE on bone turnover and BMD in healthy adolescent women and, in addition, to ascertain the influence of body mass index (BMI) and exercise on these indices of bone metabolism. METHODS: We recruited to the study 92 healthy girls aged between 16 and 19. They were divided into three groups. Participants in the first two groups used an OC with either 15 or 30 microg ethinyloestradiol (EE), whereas those in the third group used no hormonal contraception. Bone mineral density (BMD) and bone turnover markers were measured before and after 12 months of treatment. RESULTS: The BMD values of the total hip in females using the OC containing 30 microg EE was 0.912 g/cm(2) at baseline and 0.918 g/cm(2) after one year; in females using the OC containing 15 microg EE the corresponding values were 0.888 g/cm(2) and 0.895 g/cm(2) whereas in females who used no contraception BMD values were 0.942 g/cm(2) and 0.949 g/cm(2), respectively. The changes were statistically insignificant. Levels of osteocalcin and CTX had decreased after one year in all groups, but not statistically significantly so. CONCLUSION: Low dose and ultra-low dose oral contraceptives did not significantly differ in their effects on bone mineral density or bone turnover markers in adolescent girls aged 16-19. PMID- 19565419 TI - Sexual and reproductive health of adolescents in Belgium, the Czech Republic, Estonia and Portugal. AB - OBJECTIVES: This pilot study, within the REPROSTAT 2 Project, aimed at investigating the sexual and reproductive health (SRH) of school-based youths in four countries of the European Union. METHODS: Students of either sex, aged between 16 and 19 years and entering grade 10, 11 or 12 during the 2005-2006 school year participated in a cross sectional survey. A structured questionnaire was administered to a convenience sample of students from selected schools in Belgium, the Czech Republic, Estonia and Portugal. RESULTS: In each country, more than three quarters of respondents had previously had a boyfriend or girlfriend and almost half had heterosexual intercourse. More than 85% of sexually experienced youths in each country had used contraception at coital debut. Mean age at coital debut varied between 15.2 (Belgium) and 16.4 years (Czech Republic). While 51% of respondents in Estonia knew of Chlamydia trachomatis infection, the corresponding figure in Portugal was only 12%. CONCLUSIONS: Sexual activity is common among school-going youths in all four countries. This highlights the need for Member States to systematically collect indicators of SRH in this population. Use of a common measurement framework can inform the establishment of common targets for joint initiatives such as reducing teenage pregnancy and STIs in adolescents. PMID- 19565420 TI - Pharmacokinetics of an oral contraceptive containing oestradiol valerate and dienogest. AB - OBJECTIVE: To evaluate the pharmacokinetics of a combined oral contraceptive (OC) containing oestradiol valerate/dienogest (E2V/DNG) administered according to a four-phasic dosing regimen with an oestrogen step-down and a progestin step-up over 26 days of active treatment. METHODS: This Phase I, open-label study included healthy women aged 18-50 years. Treatment consisted of the administration of E2V 3 mg for 2 days, E2V 2 mg/DNG 2 mg for 5 days, E2V 2 mg/DNG 3 mg for 17 days, E2V 1 mg for 2 days, and placebo for 2 days. RESULTS: Pharmacokinetic data were analysed in 15 women. Stable E2 concentrations were maintained throughout the study. Minimum mean serum E2 levels were 33.6-64.7 pg/ml during E2V administration. The ratio of oestrone:E2 in serum was approximately 5:1. Minimum mean serum DNG levels were 6.8-15.1 ng/ml during DNG administration. Minimum concentrations of DNG increased only slightly during each phase of the regimen during which DNG was being administered. On day 24 the geometric mean C(max), C(ave) and t((1/2)) of DNG were 82.9 ng/ml, 33.7 ng/ml and 12.2 hours, respectively; the median t(max) was 1.5 hours. Serum sex hormone binding globulin concentrations increased by 40% (within the normal range). Cortisol binding-globulin levels remained almost unchanged. Treatment was well tolerated. CONCLUSIONS: Treatment with an OC containing E2V and DNG was well tolerated and was associated with stable E2 concentrations over 28 days. The pharmacokinetics of DNG were consistent with previous findings. Minimum serum concentrations of DNG increased only slightly during phases of the regimen during which DNG was administered. PMID- 19565421 TI - Antifertility activity of leaves of Plumbago zeylanica Linn. in female albino rats. AB - OBJECTIVES: To evaluate the antifertility properties of extracts of leaves of Plumbago zeylanica Linn. (Plumbaginaceae). METHODS: The effects of petroleum ether, chloroform, acetone, ethanol and aqueous extracts of the leaves of P. zeylanica on the estrous cycle of rats were studied at two dose levels, namely, 200 and 400 mg/kg. The effective extracts were assessed with regard to their oestrogenic activity in the same species. RESULTS: The acetone and ethanol extracts were most effective in interrupting the estrous cycle of the rats (p < 0.05). The animals exhibited a prolonged diestrous stage of the estrous cycle corresponding to a temporary inhibition of ovulation. The antiovulatory activity was reversible on discontinuation of treatment. Both extracts showed significant oestrogenic and anti-oestrogenic activity (p < 0.05). CONCLUSIONS: The acetone and ethanolic extracts of leaves of P. zeylanica have an antifertility activity. PMID- 19565422 TI - Comparison of the effects of the levonorgestrel-releasing intrauterine system (Mirena) and depot-medroxyprogesterone acetate (Depo-Provera) on subendometrial microvascularisation and uterine artery blood flow. AB - OBJECTIVE: To compare the effects of the levonorgestrel-intrauterine system (LNG IUS) and depot-medroxyprogesterone acetate (DMPA) on subendometrial vascularisation and uterine artery blood flow. STUDY DESIGN: The trial compared 25 patients wearing a LNG-IUS and 25 patients receiving DMPA. The subendometrial blood flow was evaluated using power Doppler analysis; the pulsatility index (PI) and resistance index (RI) in the uterine artery and spiral arteries were evaluated with spectral Doppler. RESULTS: The PI and RI of the uterine artery and endometrial thickness did not differ significantly between the two groups. The subendometrial vascularisation was lower and the PI in the spiral arteries was higher in DMPA-users. CONCLUSION: In spite of a similar prevalence of bleeding disturbances, the effects of DMPA and the LNG-IUS on the subendometrial microvascularisation differ. PMID- 19565425 TI - Small for gestational age neonates--are we missing some by only using standard population growth standards and does it matter? AB - OBJECTIVES: Birth weight below the 10th percentile on growth charts based on population norms or small for gestational age (SGA) is associated with adverse perinatal outcome. We compare the association among neonatal mortality, morbidity and SGA as determined by currently used percentiles based on population standards (method A) and customised birth weight percentiles (method B). METHODS: Study of outcomes of SGA neonates from a cohort of 6125 consecutive pregnancies, analysed in function of the method (A or B) used to diagnose SGA. RESULTS: There were 831 SGA infants, 84% were born at term and the majority were of European origin (61.4%). By method A, there was a sixfold decrease in the prevalence of SGA babies (2.3%) compared with method B (13.5%). After correcting for prematurity, SGA infants diagnosed by method B alone (not identified by standard population method) still constituted a high proportion of infants with hypothermia and hypoglycaemia (p < 0.05). CONCLUSION: Customised charts identified six times more SGA infants than standard charts. These infants, considered appropriately grown by standard charts, are at a significantly higher risk of morbidity. PMID- 19565426 TI - Neonatal brain imaging and the identification of metabolic acidemia and hypoxic ischemic encephalopathy. AB - OBJECTIVE: To determine the precision with which intrapartum metabolic acidemia and hypoxic-ischemic encephalopathy (HIE) in term and near-term infants can be identified by neonatal brain imaging. STUDY DESIGN: This is a case-control study whose inclusion criteria were neonates born at > or =34 weeks gestation with a cord gas at delivery, suspected neurological abnormalities, and computed tomography (CT) or magnetic resonance (MR) imaging of the brain. Neonates with chromosomal and major congenital malformations were excluded. Brain imaging for neonates with and without metabolic acidemia (pH < 7.0 and base deficit > 12 mM) at birth and HIE were retrospectively reviewed by a neuroradiologist blinded to their clinical course and compared. RESULTS: There were 54 neonates admitted to the NICU at a single university hospital between 1992 and 2006 that met these inclusion criteria of which 27 had metabolic acidemia at birth. There were 16 diagnosed clinically as having HIE at the time of neonatal discharge, 13 from the acidemic group and 3 from the nonacidemic group. Radiological signs of basal ganglia injury were significantly more common in neonates with metabolic acidemia (29.6%, 3.7%, p = 0.02) and HIE (37.5%, 7.9%, p = 0.01). Logistic regression corrected for gestational age showed that radiological signs of basal ganglia injury could identify the presence of HIE with area under the ROC curve of 0.71, sensitivity 37.5%, specificity 92.1%, positive predictive value 66.7%, and negative predictive value of 77.8%. CONCLUSION: Radiological signs of basal ganglia injury on early neonatal imaging are associated with metabolic acidemia and HIE, but is not precise enough to serve as a gold standard in the identification of these conditions. PMID- 19565427 TI - Epidemiology of preterm birth in Omdurman Maternity hospital, Sudan. AB - OBJECTIVE: To investigate the prevalence and to identify risk factors for live singleton preterm birth in Omdurman Maternity hospital, Sudan. METHODS: Case control study. Odds ratios were adjusted for the confounding factors using multiple logistic regression models. RESULTS: Among 3240 live birth at maternity hospital during the 3 months period of the study, 126 (3.8%) were a live preterm birth. About 99 live preterm births at a mean (SD) of 32.7 (2.2) gestational weeks were included in the final analyses. About 80 (81.0%) of these 99 preterm births were spontaneous preterm births and the rest 19 (19.0%) were medically indicated preterm births. Univariate and Multivariate analyses showed that those women who had history of miscarriage, preterm birth, cesarean delivery, dental maneuver, and vaginal bleeding in the index pregnancy, both low body mass index (BMI) (<25) and high BMI (>30) and those women who had short interpregancy interval were at higher risk for preterm birth. CONCLUSIONS: The study showed that spontaneous preterm birth predominates in this setting. Preterm birth has a tendency to recur. Infections (periodontal) and nutrition were the risk factors for preterm birth. More research is needed especially in the area of nutrition and oral hygiene. PMID- 19565428 TI - Inhibition of evaluative and perceptual odour-taste learning by attention to the stimulus elements. AB - Pairing of odours and tastes in solution results in the odour taking on the taste's hedonic and perceptual properties. Theoretical accounts of such conditioning propose that the stimulus elements form a configuration with hedonic and/or perceptual properties. One implication of this account is that configural binding may be inhibited by allocation of attention to the individual elements during conditioning (analytical attention). This was tested here by training participants to attend analytically or synthetically during pairing of odours with either sucrose or water. Differential conditioning effects for sucrose- and water-paired odours were seen only in the synthetic group, with sucrose-paired odours increasing in both smelled sweetness and liking. These data support configural models as explanations for these forms of associative learning and emphasize the role of top-down processes in mediating transfer of perceptual and hedonic properties from tastes to odours. PMID- 19565429 TI - An investigation into the online processing of counterfactual and indicative conditionals. AB - The ability to represent conditional information is central to human cognition. In two self-paced reading experiments we investigated how readers process counterfactual conditionals (e.g., If Darren had been athletic, he could probably have played on the rugby team) and indicative conditionals (e.g., If Darren is athletic, he probably plays on the rugby team). In Experiment 1 we focused on how readers process counterfactual conditional sentences. We found that processing of the antecedent of counterfactual conditionals was rapidly constrained by prior context (i.e., knowing whether Darren was or was not athletic). A reading-time penalty was observed for the critical region of text comprising the last word of the antecedent and the first word of the consequent when the information in the antecedent did not fit with prior context. In Experiment 2 we contrasted counterfactual conditionals with indicative conditionals. For counterfactual conditionals we found the same effect on the critical region as we found in Experiment 1. In contrast, however, we found no evidence that processing of the antecedent of indicative conditionals was constrained by prior context. For indicative conditionals (but not for counterfactual conditionals), the results we report are consistent with the suppositional account of conditionals. We propose that current theories of conditionals need to be able to account for online processing differences between indicative and counterfactual conditionals. PMID- 19565430 TI - Impaired task switching performance in children with dyslexia but not in children with autism. AB - Problems with cognitive control in both autism and dyslexia have already been reported in different studies. The present study specifically examined task switching performance in children with autism and dyslexia. For this purpose, a multiple-trial paradigm was used with cues for colour- and shape-matching tasks presented before a run of trials. The cue could imply a task switch (when the cue changed the task) or a task repetition (when the cue did not change the task). Both reaction times and error rates were measured for switching, restarting, and general task performance. Participants were children with autism (24) and with dyslexia (25) and healthy controls (27) with normal IQ and ages from 12 to 18 years. The main finding was that while similar switching performance was observed between children with autism and the healthy controls, children with dyslexia showed a significant switch-specific delay relative to both healthy controls and children with autism. Furthermore, no deficit in restarting performance was observed for any of the two patient groups. Finally, additional evidence is provided for a more general deficit in information processing in dyslexia. Our data suggest that children with autism are able to switch between tasks in a similar way as do normally developing children as long as the tasks are unambiguously specified. Furthermore, the data imply switch-specific deficits in dyslexia additionally to the deficits in general information processing already reported in the literature. The implications of our data are further discussed in relation to the interpretation of the Wisconsin Card Sorting Test. PMID- 19565431 TI - Finding meaning in art: preferred levels of ambiguity in art appreciation. AB - Uncertainty is typically not desirable in everyday experiences, but uncertainty in the form of ambiguity may be a defining feature of aesthetic experiences of modern art. In this study, we examined different hypotheses concerning the quantity and quality of information appreciated in art. Artworks were shown together with auditorily presented statements. We tested whether the amount of information, the amount of matching information, or the proportion of matching to nonmatching statements apparent in a picture (levels of ambiguity) affect liking and interestingness. Only the levels of ambiguity predicted differences in the two dependent variables. These findings reveal that ambiguity is an important determinant of aesthetic appreciation and that a certain level of ambiguity is appreciable. PMID- 19565432 TI - Modulation of spatial Stroop by object-based attention but not by space-based attention. AB - Earlier studies have shown that the spatial Stroop effect systematically decreases when a peripheral precue is presented at the same location as the target, compared to an uncued location condition. In this study, two experiments were conducted to explore whether the cueing modulation of spatial Stroop is object based and/or space based. In Experiment 1, we found evidence favouring the view that the cueing modulation of the spatial Stroop effect is entirely object based, as no differences were found in conflict reduction for the same-location and same-object conditions. In Experiment 2, the cue was predictive, and a similar object-based modulation of spatial Stroop was still observed. However, the direction of such modulation was affected by the rectangles' orientation. Overall, the pattern of results obtained favours the object-integration (Lupianez & Milliken, 1999; Lupianez, Milliken, Solano, Weaver, & Tipper, 2001) and referential-coding accounts (Danziger, Kingstone, & Ward, 2001) and seems to provide evidence against the attention-shift account (Rubichi, Nicoletti, Iani, & Umilta, 1997; Stoffer, 1991). PMID- 19565433 TI - The effect of recombinant human erythropoietin on the development of retinopathy of prematurity. AB - In addition to its hematopoietic effects, erythropoietin causes an increased release of endothelin-1 and the stimulation of angiogenesis and thereby it may have possible role in development of retinopathy of prematurity (ROP). Our objective was to determine if an association exists between recombinant human erythropoietin (rhEPO) treatment and the development of ROP. Our case-control study involved 85 very low birthweight infants with birthweights <1500 g born during 2003 and 2004. All the infants were divided into two groups on the basis of whether they got rhEPO or not. The rhEPO was given at the dose of 200 to 250 units/kg/dose three times a week for 10 doses. Further duration of rhEPO therapy was decided on the basis of the clinical response. Ophthalmological examinations were done at the age of 5 to 6 weeks and were repeated 1 to 4 weeks after the first examination according to the severity of the ROP disease during their in hospital stay. Of 85 infants, 56 (66%) received rhEPO and 29 (34%) did not. In the rhEPO-treated group, 12 infants (21%) had ROP; in the non-rhEPO group, 11 infants (38%) developed ROP. This difference is not statistically significant (odds ratio = 2.63; P = 0.10). There was no correlation between the use of rhEPO and the stage of ROP (random sample = -0.01; P = 0.89). There was no significant difference in the incidence of plus, prethreshold, or threshold disease and the treatment required for ROP between the rhEPO-treated and the nontreated group. The study showed there is no significant difference in the incidence and severity of ROP between the rhEPO-treated and nontreated group. PMID- 19565434 TI - Pregnancy outcome of patients with schizophrenia. AB - We sought to identify whether schizophrenia during pregnancy is associated with adverse perinatal outcomes. A population-based study comparing women with and without schizophrenia and schizoaffective disorders was performed. Stratified analysis using multiple logistic regression models was performed to control for confounders. During the study period, there were 186,554 deliveries, of which 97 occurred in patients with schizophrenia and schizoaffective disorders. The schizophrenic patients were significantly older (mean age 30.6 versus 28.6, P = 0.001), with higher prevalence of diabetes mellitus as compared with the comparison group (13.4% versus 6.7%, P = 0.009). The need for induction and augmentation of delivery, congenital malformations, and low birth weight (<2500 g) were significantly increased among schizophrenic patients. No significant differences were noted between the groups regarding labor complications such as cesarean delivery (16.5% versus 13.2%, P = 0.337) and placenta previa and placental abruption (1% versus 4%, P = 0.333 and 1% versus 0.7%, P = 0.51, respectively). Using a multivariable logistic regression model, schizophrenia and schizoaffective disorders during pregnancy were independent risk factors for congenital malformations (odds ratio 2.1; 95% confidence interval, 1.1 to 3.9, P = 0.027). Schizophrenia and schizoaffective disorders are independent risk factors for congenital malformations. PMID- 19565435 TI - Shoulder dystocia: comparison of the ACOG practice bulletin with another national guideline. AB - Our objective was to compare national guidelines regarding shoulder dystocia. Along with the American College of Obstetricians and Gynecologists (ACOG) practice bulletin on shoulder dystocia, guidelines from England, Canada, Australia, and New Zealand were reviewed. The Royal College of Obstetricians and Gynaecologists (RCOG) guideline agrees with the ACOG definition of shoulder dystocia, but there are variances in the management of suspected macrosomia and resolution of impacted shoulders. How recommendations are categorized differ also. Only 53% (20 of 38) of eligible references are cited by both publications. The two national guidelines on shoulder dystocia have differences and disagreements with each other, raising concerns about how the literature is synthesized and which is more comprehensive. PMID- 19565436 TI - Report on the 11- to 13+6-week ultrasound evaluation as a screening test for trisomy 21 in singleton pregnancies. AB - We sought to update the value of the 11- to 13 (+6)-week ultrasound evaluation for prenatal screening of trisomy 21. An English literature search was conducted for pertinent original research articles, review articles, and websites related to the 11- to 13 (+6)-week ultrasound evaluation. Reports from prenatal diagnostic units published since 1990 that included two- and three-dimensional ultrasound and Doppler hemodynamic evaluations for prenatal screening of trisomy 21 between weeks 10 and 13 (+6) of gestation were selected. Alternative screening methods were identified, and clinical information concerning each of them was extracted, integrated into tables, and then synthesized to provide meaningful clinical interpretations. Finally, mechanisms that could lead to improvements in the efficiency of these screening tests are discussed. Two-dimensional measurements of the fetal nuchal translucency play a key role in screening for trisomy 21. Evaluations of the nasal bone, ductus venosus, tricuspid valve function, frontomaxillary facial angle, hyperechogenic bowel, intracardiac echogenic focus, and renal pelvis fullness can become part of the 11- to 13 (+6) week screening test if the imaging protocols are standardized. The role of 3-D ultrasound during the 11- to 13 (+6)-week ultrasound evaluations is under investigation. PMID- 19565437 TI - Antimicrobial, antiproliferative, cytotoxic, and tau inhibitory activity of rubellins and caeruleoramularin produced by the phytopathogenic fungus Ramularia collo-cygni. AB - Photodynamically active anthraquinone derivatives produced by the phytopathogenic fungus Ramularia collo-cygni are known to cause a barley leaf-spot disease, but data about light-dependent and independent bioactivity have been sparse to date. We therefore conducted for the first time a broad bioactivity profiling of rubellins B, C, D, and E and caeruleoramularin. Antibacterial but not antiviral activity is reported with light-dependent increase. Furthermore, when tested without illumination, compounds exerted antiproliferative and cytotoxic activity in a series of human tumor cell lines. Inhibition of tau protein assembly was observed as well. PMID- 19565438 TI - [How are religiosity, psychological distress and integration interrelated? A comparative study of the religiosity in Turkish/Kurdish migrants in comparison to Germans]. AB - OBJECTIVE: The current study aimed to assess the association between religious involvement, psychological distress and various aspects of integration in a population of Turkish/Kurdish migrants in primary care. METHODS: We compared the group of Turkish/Kurdish migrants (n = 179, age m = 39.4 years, 65.0 % female) with German dermatological patients (n = 184, age m = 43.1 years, 60.9 % female) with regard to their religiosity. RESULTS: On average the migrants lived in Germany for 24.98 years (SD = 8.90). Compared to the German patients, they showed higher religious involvement and participation. The migrants were highly significant more religious (p < 0.001) than the German patients. Contrary to expectation, no significant correlations between measures of religiosity and psychological distress were found for the Turkish migrant group. Further, inverse relationships between measures of religiosity and measures of integration resulted. Migrants who were more acculturated showed less religiosity. CONCLUSIONS: The religious affiliation of the examined group was islamic. It remains open if the inverse association between religiosity and integration would be the same also for other religions. PMID- 19565439 TI - [Is patient's satisfaction with inpatient depression treatment related to their German language proficiency?]. AB - OBJECTIVE: To analyze patient satisfaction in inpatient depression treatment depending on German language proficiency. METHODS: Documentation of patients in 24 psychiatric hospitals in Baden-Wurttemberg (South Germany). The following three groups were matched according to age, gender, and education: native German speakers, non-native speakers with good German proficiency, and non-native speakers with limited German proficiency (67 in each group). RESULTS: The number of unsatisfied patients was four times higher in the group of non-native speakers with limited German proficiency (24 %) than in the native German speakers group (6,5 %) and more then twice as high as in the group of non-native speakers with good German proficiency (10.2 %). Patient satisfaction was lower in the group of non-native speakers with limited German proficiency independent of treatment outcome than in both other groups, whereas in both groups with good German proficiency, satisfaction was associated with treatment outcome. CONCLUSIONS: Good linguistic understanding is more important for patient satisfaction in inpatient depression treatment than treatment outcome. PMID- 19565440 TI - [Shared psychotic disorder (Folie a deux) and Huntington's disease]. AB - INTRODUCTION: Folie a deux describes a rare psychiatric disorder of mostly closely related persons. The main feature of this disease is the unconditional adoption of the delusions of the primarily diseased person by the second person. This disturbance most frequently originates from a paranoid schizophrenia. METHOD: Our case is the first published description of a married couple with shared psychotic disorders which were caused by a genetically verified Chorea Huntington of the husband. RESULTS: The symptoms of the wife quickly declined after spatial separation from her primarily diseased husband. Thus, in her case it was assumed that she suffered from a Folie imposee which is a sub form of the Folie a deux. DISCUSSION: Our case report demonstrates that not only a paranoid schizophrenia but also an organic psychosis may cause this very interesting form of shared psychotic disorder. PMID- 19565441 TI - [Automutilation with indistinct underlying disease--difficulties of care in the community]. AB - INTRODUCTION: Self-mutilation is well-known in various psychiatric diseases and represents a challenge for forensic pathologists as regards the differentiation of accidental versus self-inflicted injuries, especially when a criminal charge is likely to emerge. METHOD: A case of extraordinarily severe self-inflicted injuries is presented as well as the related implications concerning clinical and ambulatory care for patients, whose underlying motivation remains unknown. RESULTS: A 60 year-old worker exhibited an open scull fracture and 14 abdominal wounds with protrusion of small intestine, from which a part of 160 cm length was completely removed beforehand. The patient claimed the wounds having been caused accidentally. When examined by a psychiatrist, disturbances in orientation, unrest and ill concentration were found, but no symptoms related to psychosis, depression or suicidal behaviour. DISCUSSION: Although the need for more detailed examination seemed to be clearly evident, the medical authorities did not take action. Consequently, no further psychiatric treatment was initiated. The implications of this case on the background of the literature are discussed. PMID- 19565442 TI - Predictors of postoperative stricture after esophageal endoscopic submucosal dissection for superficial squamous cell neoplasms. AB - BACKGROUND AND STUDY AIMS: Although endoscopic submucosal dissection (ESD) is becoming accepted as an established treatment for superficial esophageal squamous cell neoplasms, the risks for developing postoperative stricture have not been elucidated. PATIENTS AND METHODS: This was a retrospective study at a single institution. From January 2002 to October 2008, 65 patients with high-grade intraepithelial neoplasms (HGINs) or m2 carcinomas treated by ESD were enrolled. Predictors of postoperative stricture were investigated by comparing results from 11 patients who developed strictures with those from 54 patients who did not. RESULTS: Significant differences between the two groups were observed in longitudinal diameter (45.0 +/- 15.9 mm vs. 31.5 +/- 13.6 mm) and circumferential diameter (37.2 +/- 8.6 mm vs. 26.8 +/- 9.7 mm) of the resected specimens, and the proportion of extension to the whole circumference of the lumen (< 1 / 2/ > 1 / 2/ > 3 / 4 : 2 / 4 / 5 vs. 40 / 13 / 1), histologic depth (HGIN/m2 : 2 / 9 vs. 41 / 13), and procedure time (85.6 +/- 42.8 minutes vs. 53.3 +/- 30.1 minutes). Multivariate analysis revealed that circumferential extension of > 3 / 4 (odds ration [OR]: 44.2; 95 % confidence interval [CI]: 4.4 - 443.6) and histologic depth to m2 (OR: 14.2; 95 %CI: 2.7 - 74.2) are reliable risk factors. Subanalysis for each category by combinations of these risk factors revealed that patients with lesions in > 3 / 4 of the circumferential area were associated with a high rate of postoperative stricture. By contrast, patients with HGIN lesions in < 3 / 4 extension have no probability of postoperative strictures. Additionally, subanalysis of patients with m2 lesions in < 3 / 4 circumferential extension revealed that circumferential diameter can be a reliable predictor for postoperative stricture. CONCLUSIONS: Circumferential extension and histologic depth are the reliable risk factors for postoperative strictures. In combination with circumferential diameter, we can perform effective and appropriate preventive balloon dilatations after esophageal ESD. PMID- 19565443 TI - [Preoperative evaluation of patients for elective surgery]. AB - Perioperative risk analysis and risk reduction is the main goal of every preoperative evaluation. Exercise tolerance is a key determinant of patient risk. Preoperative tests should only be ordered based on defined indications and relevant findings. Empathy towards the patient as well as good information is crucial. "Remote premedication" is a new method for low risk patient groups with phone and internet access. Based on modern information technology, this new method can be more efficient while preserving information flow and patient satisfaction. PMID- 19565444 TI - [The preoperative anaesthetic visit]. AB - Anaesthetists often visit their patients in exceptional situations characterised by preoperative anxiety or distress. Therefore, even brief contact with the patient can be considered intense and meaningful. The initial preoperative anaesthetic visit is the beginning of the relationship between patient and anaesthetist, and should help to explain the planned anaesthetic technique. Preoperative anaesthetic visits are intense and last for 20 minutes on average. They should assert a professional approach to the patient's emotions, particularly to preoperative anxiety, and a structured and clear collection of information including the past history of the patient. These visits should also provide information about the anaesthesia itself and instructions for the patient with respect to the perioperative period. Communication about the side effects and risks of anaesthetic techniques, and the discussion of potential alternatives are mandatory. Worldwide, courts of law increasingly require a documented discussion between the anaesthetist and patient based on risk-benefit evidence. Today, there is in general a shift away from decisions made solely by physicians, reflecting an increased respect for the autonomy of the patient towards a model of shared decision-making and informed choice. Ideally, the preoperative visit follows the four key habits of highly effective clinicians, i.e., to rapidly establish a rapport with the patient and provide an agenda for the visit, to explore the patient's perspectives and expectations, to demonstrate empathy, and to focus on the end of the visit with providing information and including the patient in the decision-making process. Visits are then concluded upon obtaining informed consent from the patient. PMID- 19565445 TI - [Perioperative management of chronic medication: to withhold, continue or intensify?]. AB - All patients should undergo surgical procedures in the most stable and favourable condition with a continuation of chronic medication. Accordingly, this medication should be continued until the day before surgery in most patients. The preoperative period should be used to evaluate the completeness and quality of the medical treatment. Are all patient with cardiological indications for beta receptor blocking agents, ACE-inhibitors and statins treated with these drugs? Is diabetes being adequately treated? Are the doses of the medication for the treatment of thyroid dysfunction within the optimal range? If there is room for improvement, adjustments should be done cautiously and well in advance. In contrast, there are no recognised situations in which a medication should be started preoperatively in the absence of a pre-established chronic indication. Management of antiplatelet agents and cumarins requires careful consideration. The hazards of continuing and withholding treatment must be evaluated. In patients who have suffered a cardiovascular event and/or have undergone (coronary) revascularisation, elective surgery must be delayed for weeks or months depending on the circumstances and the devices used. PMID- 19565446 TI - [Preoperative check-up of patients with neurological disease]. AB - Often, patients with neurological diseases need to undergo surgical procedures. The most frequent are orthopaedic surgery (tenotomies, corrections of articulations), urological procedures (for instance, sphincter surgery for urinary incontinence), ENT surgery (tracheotomy), or plastic surgery for closure of decubitus ulcers. Preoperatively, these patients need a careful check-up including a thorough drug history. Specific diagnostic interventions may be indicated, for instance, lung functions tests in patients with muscular weakness undergoing thoracic or upper abdominal surgery. A close collaboration between general practitioner and anaesthesiologist is warranted. It cannot be excluded that symptoms of the neurological disease will deteriorate postoperatively due to anaesthesia, surgery or perioperative stress. Medication that is used to control symptoms of a neurological disease should not be stopped in the perioperative period. PMID- 19565447 TI - [Preoperative preparation of patients with cardiovascular disease for noncardiac surgery]. AB - Preoperative preparation of patients with cardiovascular disease is best initiated by the general practitioner. Updated Guidelines on Perioperative Cardiovascular Evaluation and Care for Noncardiac Surgery have been published by the American Heart Association und American College of Cardiology (2007). Individual cardiac evaluation must take into account active cardiac conditions, functional capacity, additional clinical risk factors and surgical risk. Stable, asymptomatic patients with normal functional capacity can proceed to elective anesthesia and surgery without further cardiac evaluation. Active cardiac conditions require evaluation and treatment by a cardiology service prior to elective surgery. In stable patients with poor (<4 metabolic equivalents, MET) or unknown functional capacity and clinical risk factors, who are scheduled for intermediate- or high-risk surgery, further cardiac evaluation and preparation is to be considered. Established indicated beta blocker and statin medication is to be continued; timely institution of beta blocker medication (target heart rate, <65 bpm) may be required depending on the risk of surgery, the presence of coronary heart disease, and the number of clinical risk factors present. Following percutaneous coronary intervention, specific waiting periods are required prior to elective surgery. In patients on antiplatelet therapy, the risk of stopping it should be weighed against the benefit of reduction in bleeding complications from the planned surgery. PMID- 19565448 TI - [Airways and respiratory function: preoperative screening--anaesthetic and intensive care aspects and considerations]. AB - The risk factors for respiratory complications can broadly be classified as patient-related or procedure-related factors. Knowledge about pathophysiological and pharmacological aspects of the patients' condition is essential for their safe care during preoperative assessment, intra-operative management and, if necessary, postoperative intensive care. A good understanding of the risk factors leading to the development of postoperative respiratory complications should result in the implementation of perioperative strategies designed to prevent their occurrence and reduce severity. Special information regarding airway and lung aspects as well as cases involving different types of anaesthesia and surgical procedures are also discussed. PMID- 19565449 TI - [Preoperative management of patients with liver disease]. AB - Patients undergoing liver surgery due to benign or malignant liver tumors are evaluated in a multidisciplinary way: surgeons, gastroenterologists, anesthesiologists, intensive care physicians, radiologists, oncologists and pathologists are involved in the perioperative evaluation. Surgical know-how, combined with a broad spectrum of diagnostic and therapeutic options stand for a high degree of safety and effectiveness of perioperative treatment. The general physical status of the patient, the type and extend of liver disease, as well as the complexity of liver resection have significant impact on perioperative morbidity and mortality. Appropriate preoperative evaluation and preparation of the patient is therefore of utmost importance. This allows for adequate risk assessment and further helps to define risk management strategies. PMID- 19565450 TI - [The preanaesthetic care of patients with renal disorder]. AB - The preoperative evaluation of patients with renal disease does not begin with the anaesthesiologist's visit, but with the evaluation and care given by the primary physician. To decrease postoperative morbidity and mortality, an optimal preoperative preparation is essential. The kidney is highly sensitive to perioperative hypoperfusion. Basically the medullary oxygenation with its high rate of oxygen extraction (about 79% of the delivered oxygen) is vulnerable to hypoxia, which may lead to acute tubular necrosis. The main goal of perioperative care is to avoid hypoperfusion and therefore hypoxia of the kidney. A perioperative equalized intravascular fluid balance (normovolaemia) seems to be the most effective strategy to prevent postoperative renal dysfunction. On the other side, the anaesthesia management depends on the renal function. The dosage of anaesthetics must be adapted to an impaired renal capacity. Accurate assessment of the renal function relies on laboratory determinations. The most useful laboratory tests are those related to the glomerular filtration rate (GFR), which represents the renal function. The measurement of the creatinine clearance is the most precise method available for clinically assessing overall renal function (really GFR). Measurement of creatinine clearance by 24-hour urine collection is uncomfortable and highly error-prone. For the estimation of GFR, the use of the Cockroft-Gault or the MDRD (Modification of Diet in Renal Disease) formula is recommended. The renal impairment is grouped according to the glomerular function, estimated by the named formula. Other important parameters to estimate the effects of an advanced renal dysfunction are the haemoglobin and the serum concentration of potassium and sodium. If possible, elective operative interventions should be carried out during a stable phase of the disease. Appropriate treatment of concomitant symptoms, such as hypertonia, is essential and contributes to a reduction of post operative renal dysfunction. Timely consultation with the responsible anaesthesist can prevent unnecessary delay of the intervention. PMID- 19565451 TI - [Preoperative evaluation of patients with endocrine diseases]. AB - Patients with endocrine diseases are candidates for an early anaesthesiologic evaluation. This is especially important for diseases with a high potential for preoperative optimization of therapeutic options, as diabetes, morbid obesity and thyroid dysfunction. For rare diseases, early anaesthesiologic evaluation allows the anaesthetic team to gather specific information, which is not available on an overnight basis. PMID- 19565452 TI - [Management and preoperative evaluation for anesthesia of the pregnant woman]. PMID- 19565453 TI - [Livestock owners with specifically increased disease awareness for Bluetongue: A new approach to disease surveillance]. AB - For the first time in Switzerland, specifically trained livestock owners were included in a national disease surveillance program by the Federal Veterinary Office. A questionnaire on data about clinical and epidemiological aspects of Bluetongue Disease (BT) as well as on herd management was completed by 26 sheep owners three months after they had attended a training course about BT. The control group, consisted of 264 randomly selected sheep and cattle owners who had not visited a training course. Results showed that disease awareness for BT after attending the training course was considerably increased. This was especially evident in the better knowledge of the participants about the great number of possible symptoms. Training courses with the objective of increased disease awareness of livestock owners are an efficient, cost-effective instrument in control programs for exotic diseases. PMID- 19565454 TI - Stenotrophomonas maltophilia isolated from the airways of animals with chronic respiratory disease. AB - Stenotrophomonas maltophilia (S. maltophilia) is a nonfermentative bacterium, which is naturally resistant against a panel of commonly-used antibiotics. It is frequently isolated from humans with chronic respiratory disease, e.g. cystic fibrosis or chronic obstructive pulmonary disease. In veterinary medicine S. maltophilia is perceived to be a mere coloniser. We herewith report 7 strains of S. maltophilia isolated from animals, of which 5 strains were harvested from 3 horses, a dog and a cat with chronic respiratory disease. The dog isolate showed resistance to trimethoprim / sulphamethoxazole, which was confirmed by detection of the sul 1 gene. Analysis with pulsed field gel electrophoresis revealed that 2 horses, which were boarded in the same clinic but two years apart, harboured the same strain of S. maltophilia. This is indicative of a hospital acquired colonisation / infection, which contradicts involvement in the pre-existing chronic disease. PMID- 19565455 TI - [Raw milk-associated Staphylococcus aureus intoxication in children]. AB - Four hours after the consumption of raw goat milk, three Swiss children came down with emesis and diarrhea in July 2008. First investigations showed that the milk originated from a goat suffering from clinical mastitis (Staphylococcus aureus). In the milk sample from the untreated left udder, Staphylococcus aureus counts reached 5.0 x 10(7) CFU ml(-1). By PCR, the gene for the staphylococcal enterotoxin D was found in isolated strains. The consumption of raw milk is rarely associated with Staphylococcus aureus intoxications. Due to the flora naturally present in raw milk, Staphylococcus aureus normally cannot multiply sufficiently. However, in the present case, high Staphylococcus aureus counts were already present in the milk due to the mastitis of the goat. This amount sufficed to cause a Staphylococcus aureus intoxication in the children. PMID- 19565456 TI - [Ultrasonographic and computed tomographic findings in a goat with mediastinal lymphocytic thymoma]. AB - This case report describes the clinical, sonographic, computed tomographic and pathological findings in a 9-year-old goat with mediastinal lymphocytic thymoma. The goat was referred to the Department of Farm Animals because of weight loss and dyspnoea. The lead clinical findings were increased heart rate, increased respiratory rate and heart sounds heard only on the right side. Ultrasonographic examination revealed a massive amount of fluid and an echogenic corrugated mass ventral to the lungs in the thoracic cavity on the left side. Computed tomography showed that the mass was very large and diffusely mineralised. A tentative diagnosis of mediastinal neoplasia was made, and the goat was euthanized. Postmortem examination revealed a cauliflower-like, pedunculated tumour, which occupied the entire left thoracic cavity and displaced the left lung. Based on histological evaluation, the tumour was diagnosed as a lymphocytic thymoma. PMID- 19565457 TI - [Anaplasma phagocytophilum infection in a cat in Switzerland]. AB - The following case report describes the diagnosis and therapy of a cat with an Anaplasma phagocytophilum infection. The cat from the canton of St. Gallen was presented because of lethargy and lack of appetite. The clinical symptoms established were fever and minor exsiccosis. The diagnosis of granulocytic anaplasmosis was established through microscopic evidence of inclusion bodies in neutrophil granulocytes, the detection of pathogen DNA in the blood by PCR and positive IgM and IgG antibody titers by serological testing. Following this diagnosis the cat was treated for 20 days with doxycycline. As the body temperature normalised, the activity of the cat improved while normalisation of food intake was delayed. After therapy Anaplasma phagocytophilum DNA could not be detected by PCR and a complete remission of abnormal serum chemistry and hematological parameters could be shown. PMID- 19565458 TI - [Errors and restlessness around the theme of piglet castration]. PMID- 19565459 TI - [ "One medicine - one health" and the veterinary medicine in Switzerland]. PMID- 19565463 TI - NMR quantification of trace components in complex matrices by band-selective excitation with adiabatic pulses. AB - The use of band-selective excitation with adiabatic pulses to rapidly obtain NMR spectra of trace components in the presence of strong signals is described, along with qualitative and quantitative examples from food matrices like olive oil and honey. PMID- 19565464 TI - Target registration and target positioning errors in computer-assisted neurosurgery: proposal for a standardized reporting of error assessment. AB - BACKGROUND: Assessment of errors is essential in development, testing and clinical application of computer-assisted neurosurgery. Our aim was to provide a comprehensive overview of the different methods to assess target registration error (TRE) and target positioning error (TPE) and to develop a proposal for a standardized reporting of error assessment. METHODS: A PubMed research on phantom, cadaver or clinical studies on TRE and TPE has been performed. Reporting standards have been defined according to (a) study design and evaluation methods and (b) specifications of the navigation technology. RESULTS: The proposed standardized reporting includes (a) study design (controlled, non-controlled), study type (non-anthropomorphic phantom, anthropomorphic phantom, cadaver, patient), target design, error type and subtypes, space of TPE measurement, statistics, and (b) image modality, scan parameters, tracking technology, registration procedure and targeting technique. CONCLUSIONS: Adoption of the proposed standardized reporting may help in the understanding and comparability of different accuracy reports. PMID- 19565465 TI - A systematic study of the function of the human beta-globin introns on the expression of the human coagulation factor IX in cultured Chinese hamster ovary cells. AB - BACKGROUND: Intronic sequences have the potential to improve gene expression in eukaryotes by a variety of mechanisms. In this context, human beta-globin (hBG) introns were inserted into the human factor IX (hFIX) cDNA in cytomegalovirus (CMV)-regulated plasmids. The resulting construct was then used for further expression analysis in vitro. METHODS: Seven hFIX-expressing plasmids with different combinations of the two hBG introns and the Kozak element were constructed and used for a systematic expression analysis in cultured Chinese hamster ovary (CHO) cells. In parallel, the hBG intronic sequences were analysed for the presence of possible regulatory elements. RESULTS: All the constructed plasmids resulted in transient expression of the hFIX. However, the coagulation activities varied according to the particular constructs used. Based on the hFIX antigenic assay, a wide range of variation was observed during persistent expression. The second hBG intron appears to be more effective than the first one. The expression level was further increased upon the inclusion of the Kozak element. Sequence analysis has detected several transcription factor binding (TFB) motifs in both of the introns, but with a higher frequency in the second one. CONCLUSIONS: Potentials of hBG introns as enhancer-like elements for the expression of the hFIX in cultured CHO cells and a higher activity with respect to the second hBG intron compared to the first one were demonstrated. The larger number of TFBs in the second hBG intron reflects its stronger effect. The results obtained suggest possible synergistic functions of the hBG introns and Kozak on the expression level of hFIX in vitro. PMID- 19565466 TI - Sequence determinants of thermodynamic stability in a WW domain--an all-beta sheet protein. AB - The stabilities of 66 sequence variants of the human Pin1 WW domain have been determined by equilibrium thermal denaturation experiments. All 34 residues composing the hPin1 WW three-stranded beta-sheet structure could be replaced one at a time with at least one different natural or non-natural amino acid residue without leading to an unfolded protein. Alanine substitutions at only four positions within the hPin1 WW domain lead to a partially or completely unfolded protein-in the absence of a physiological ligand. The side chains of these four residues form a conserved, partially solvent-inaccessible, continuous hydrophobic minicore comprising the N- and C-termini. Ala mutations at five other residues, three of which constitute the ligand binding patch on the concave side of the beta-sheet, significantly destabilize the hPin1 WW domain without leading to an unfolded protein. The remaining mutations affect protein stability only slightly, suggesting that only a small subset of side chain interactions within the hPin1 WW domain are mandatory for acquiring and maintaining a stable, cooperatively folded beta-sheet structure. PMID- 19565467 TI - Neuroprotective effect of parthenocissin A, a natural antioxidant and free radical scavenger, in focal cerebral ischemia of rats. AB - Neuroprotective effects of parthenocissin A (PA), a novel antioxidant and free radical scavenger, were studied in a transient middle cerebral artery (MCA) occlusion model in rats for the first time. The animals were treated intraperitoneally with PA at 2.5, 5 or 10 mg/kg, for both 30 min before MCA occlusion and 6 h after reperfusion. The MCA was occluded for 1 h in anesthetized Sprague-Dawley rats. Compared with vehicle-treated controls, MCA occluded animals treated with PA showed dose-dependent reductions in brain infarction size with improved neurological and motor outcome. Biomedical assay showed that the PA treatment suppressed lipid peroxidation and restored superoxide dismutase (SOD) activity in brain tissue. In addition, the ischemia/reperfusion (I/R) induced elevation of nitric oxide (NO) production and nitric oxide synthase (NOS) activity in brain tissue was also inhibited. Thus, PA demonstrated a neuroprotective effect in the I/R model and the beneficial effects of the compound may result from the reduction of oxidative stress and the inhibition of NO production induced by I/R. The neuroprotective effects of PA have highlighted the potential use of stilbene oligomers in stroke therapy. PMID- 19565468 TI - Contingent valuation: (still) on the road to nowhere? PMID- 19565469 TI - NMR-based metabolomics of urine for the atherosclerotic mouse model using apolipoprotein-E deficient mice. AB - NMR-based metabolomics of mouse urine was used in conjunction with the traditional staining and imaging of aortas for the characterization of disease advancement, that is, plaque formation in untreated and drug-treated apolipoprotein-E (apoE) knockout mice. The metabolomics approach with multivariate analysis was able to differentiate the captopril-treated from the untreated mice in general agreement with the staining results. Principal component analysis showed a pattern shift in both the drug-treated and untreated samples as a function of time that could possibly be explained as the effect of aging. Allantoin, a marker attributed to captopril treatment was elevated in the drug-treated mice. From partial least squares-discriminant analysis, xanthine and ascorbate were elevated in the untreated mice and were possible markers of plaque formation in the apoE knockout mice. Several additional peaks in the spectra characterizing the study endpoint were found but their respective metabolite identities were unknown. PMID- 19565470 TI - Thermal desorption counter-flow introduction atmospheric pressure chemical ionization for direct mass spectrometry of ecstasy tablets. AB - A novel approach to the analysis of ecstasy tablets by direct mass spectrometry coupled with thermal desorption (TD) and counter-flow introduction atmospheric pressure chemical ionization (CFI-APCI) is described. Analytes were thermally desorbed with a metal block heater and introduced to a CFI-APCI source with ambient air by a diaphragm pump. Water in the air was sufficient to act as the reactive reagent responsible for the generation of ions in the positive corona discharge. TD-CFI-APCI required neither a nebulizing gas nor solvent flow and the accompanying laborious optimizations. Ions generated were sent in the direction opposite to the air flow by an electric field and introduced into an ion trap mass spectrometer. The major ions corresponding to the protonated molecules ([M + H](+)) were observed with several fragment ions in full scan mass spectrometry (MS) mode. Collision-induced dissociation of protonated molecules gave characteristic product-ion mass spectra and provided identification of the analytes within 5 s. The method required neither sample pretreatment nor a chromatographic separation step. The effectiveness of the combination of TD and CFI-APCI was demonstrated by application to the direct mass spectrometric analysis of ecstasy tablets and legal pharmaceutical products. PMID- 19565471 TI - Circumferential pharyngeal reconstruction: history, critical analysis of techniques, and current therapeutic recommendations. AB - Reconstruction of circumferential pharyngeal defects following total pharyngolaryngectomy presents major challenges with respect to surgical morbidity and restoration of functional deficits, which are often made more demanding by the increasing trend to utilize primary chemoradiation protocols with surgery reserved for salvage cases. The present review evaluates the reconstructive techniques described in the literature, including historical techniques as well as more recent innovative methods. Each technique is critically appraised with particular reference to postoperative morbidity and functional rehabilitation. Treatment recommendations are made based on the available evidence. PMID- 19565472 TI - Acetyl derivate of quercetin increases the sensitivity of human leukemia cells toward apoptosis. AB - The hydroxyl groups of flavonoids are important for their bioactive functions and also prone to oxidation to quinones. To block the potential oxidation of quercetin, and generate a stronger bioactive compound, we synthesized acetyl and methyl derivatives of quercetin, 3,7,3',4'-O-tetraacetylquercetin (4Ac-Q) and 3,7,3',4'-O-tetramethylquercetin (4Me-Q), which substituted the hydroxyl groups of quercetin with acetyl or methyl groups at the 3,7,3',4' positions of quercetin, and then evaluated the ability to cause cell proliferation inhibition and apoptosis in HL-60 cells. The results revealed that 4Ac-Q and quercetin, but not 4Me-Q, significantly inhibit cell proliferation by caspase-mediated apoptosis when characterized by DNA fragmentation, activation of caspase-3 and PARP cleavage while 4Me-Q lost this ability. Interestingly, 4Ac-Q revealed stronger apoptotic activity than parent quercetin via a ROS-independent pathway. These findings provide a valuable strategy to increase the sensitivity of human leukemia HL-60 cells toward apoptosis by modifying quercetin structure. PMID- 19565473 TI - Modulation of Zn-induced hyperinsulinemia/insulin resistance in Wistar rat fed modified poultry egg(psi). AB - Excessive bioavailability of Zn causes Cu and Mg deficiencies resulting in hyperglycemia and hyperinsulinemia/insulin resistance. These defects may ameliorate if the ionic imbalance in them is corrected. In view of this, three groups of rats were included in this study. Initially, they were fed on semi synthetic equicalories basal diet containing 20 mg Zn (control, group-I), on 40 mg Zn (group-II) and 80 mg Zn/kg diet (group-III) respectively for 3 months. Thereafter, half of the rats in group-II and III were shifted on Cu and Mg enriched modified poultry egg (ME(Psi)) mixed diets (groups-IIME and IIIME) while the remaining were continued to feed on their respective diets for another 3 months completing a total of 6 months. Hyperglycemia, hyperinsulinemia, hypercortisolemia, hyperzincemia, hypercupremia and hypermagnesaemia with corresponding increase of lipid droplets in the zona fasciculate of adrenal cortex and reduction in liver glycogen content in rats of groups-II and III were recorded. These changes were linked with a rise in Zn and fall in Cu and Mg in their liver. The addition of ME(Psi) in their diets led to fall of Zn and rise in liver Cu and Mg, and fall in serum Zn, Cu and Mg resulting in the improvement of glucose disposal, increase in insulin sensitivity, reduction in lipid droplets in zona fasciculate and increase in glycogen content in the liver approaching closer to the control group-I. The data suggest that these ME(Psi) can serve as non pharmacological dietary supplement to prevent insulin resistance/hyperinsulinemia in populations who are at higher risk of diabetes mellitus either due to their genetic predisposition of excessive absorption and retention of Zn or due to higher Zn content in the food chain. PMID- 19565474 TI - The ERK signaling cascade--views from different subcellular compartments. AB - The extracellular signal-regulated kinase cascade is a central signaling pathway that is stimulated by various extracellular stimuli. The signals of these stimuli are then transferred by the cascade's components to a large number of targets at distinct subcellular compartments, which in turn induce and regulate a large number of cellular processes. To achieve these functions, the cascade exhibits versatile and dynamic subcellular distribution that allows proper temporal and spatial modulation of the appropriate processes. In this review, we discuss the intracellular localizations of different components of the ERK cascade, and the impact of these localizations on their activation and specificity. PMID- 19565475 TI - The safety and efficacy of a JAK inhibitor in patients with active rheumatoid arthritis: Results of a double-blind, placebo-controlled phase IIa trial of three dosage levels of CP-690,550 versus placebo. AB - OBJECTIVE: To determine the efficacy, safety, and tolerability of 3 different dosages of CP-690,550, a potent, orally active JAK inhibitor, in patients with active rheumatoid arthritis (RA) in whom methotrexate, etanercept, infliximab, or adalimumab caused an inadequate or toxic response. METHODS: Patients (n = 264) were randomized equally to receive placebo, 5 mg of CP-690,550, 15 mg of CP 690,550, or 30 mg of CP-690,550 twice daily for 6 weeks, and were followed up for an additional 6 weeks after treatment. The primary efficacy end point was the American College of Rheumatology 20% improvement criteria (ACR20) response rate at 6 weeks. RESULTS: By week 6, the ACR20 response rates were 70.5%, 81.2%, and 76.8% in the 5 mg, 15 mg, and 30 mg twice daily groups, respectively, compared with 29.2% in the placebo group (P < 0.001). Improvements in disease activity in CP-690,550-treated patients compared with placebo were seen in all treatment groups as early as week 1. ACR50 and ACR70 response rates significantly improved in all treatment groups by week 4. The most common adverse events reported were headache and nausea. The infection rate in both the 15 mg twice daily group and the 30 mg twice daily group was 30.4% (versus 26.2% in the placebo group). No opportunistic infections or deaths occurred. Increases in mean low-density lipoprotein cholesterol and high-density lipoprotein cholesterol levels, and increases in mean serum creatinine level (0.04-0.06 mg/dl) were seen in all CP 690,550 treatment arms. CONCLUSION: Our findings indicate that CP-690,550 is efficacious in the treatment of RA, resulting in rapid, statistically significant, and clinically meaningful reductions in the signs and symptoms of RA. Further studies of CP-690,550 in RA are warranted. PMID- 19565476 TI - Anti-DNA antibody induction of protein kinase C phosphorylation and fibronectin synthesis in human and murine lupus and the effect of mycophenolic acid. AB - OBJECTIVE: To examine fibronectin (FN) expression in human lupus nephritis and the effect of anti-DNA antibodies on transforming growth factor beta1 (TGFbeta1) and FN synthesis in cultured human mesangial cells. The effects of mycophenolic acid (MPA) on this pathway, and the effects of mycophenolate mofetil (MMF) treatment in (NZB x NZW)F(1)/J mice were also studied. METHODS: Immunohistochemical analyses of renal biopsy samples from patients with active diffuse proliferative lupus nephritis were performed. Cultured human mesangial cells were incubated with human polyclonal anti-DNA antibodies, with or without MPA. (NZB x NZW)F(1)/J mice with active nephritis were randomized to receive either MMF (100 mg/kg/day) or vehicle treatment for 12 weeks. RESULTS: Glomerular FN expression was increased in patients with lupus nephritis, and it colocalized with IgG deposition. Anti-DNA antibodies induced protein kinase Calpha (PKCalpha), PKCbetaI, and PKCbetaII activation, increased levels of bioactive TGFbeta1, and increased FN synthesis in human mesangial cells (P < 0.001 for each comparison versus control conditions). Pretreatment of anti-DNA antibodies with exogenous DNA reduced their cellular binding and abrogated their induction of TGFbeta1 and FN synthesis. Inhibition of PKC activation in human mesangial cells prior to anti-DNA antibody stimulation had no effect on cell proliferation, but resulted in significantly reduced antibody-mediated TGFbeta1 secretion and FN synthesis. MPA treatment down-regulated PKCalpha, PKCbetaI, and PKCbetaII phosphorylation, reduced levels of TGFbeta1 bioactivation, and decreased FN synthesis and deposition into the extracellular matrix. MMF treatment in (NZB x NZW)F(1)/J mice resulted in a reduction in glomerular IgG deposition, PKC activation, and FN expression, as well as an amelioration of proteinuria. CONCLUSION: Human polyclonal anti-DNA antibodies induce TGFbeta1 and FN synthesis in human mesangial cells through PKC activation, which is inhibited by MPA. PMID- 19565477 TI - Interleukin-6 is a significant predictor of radiographic knee osteoarthritis: The Chingford Study. AB - OBJECTIVE: There is a great need for identification of biomarkers that could improve the prediction of early osteoarthritis (OA). We undertook this study to determine whether circulating levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha), and C-reactive protein (CRP) can serve as useful markers of radiographic knee OA (RKOA) in a normal human population. METHODS: RKOA data were obtained from the cohort of the Chingford Study, a prospective population based study of healthy, middle-aged British women. The RKOA-affected status of the subjects was assessed using the Kellgren/Lawrence (K/L) grade as determined on radiographs obtained at baseline (n = 908) and at 10 years and 15 years thereafter. Serum levels of CRP, IL-6, and TNFalpha were assayed at 5, 8, and 15 years, using high-sensitivity commercial assays. A K/L grade of >or=2 in either knee was used as the outcome measure. Statistical analyses included analysis of variance for repeated measurements and logistic regression models, together with longitudinal modeling of dichotomous responses. RESULTS: During 15 years of followup, the prevalence of RKOA (K/L grade >or=2) increased from 14.7% to 48.7% (P < 0.00001 versus baseline). The body mass index (BMI) and circulating levels of CRP and IL-6 were consistently and significantly higher in subjects diagnosed as having RKOA. When multiple logistic regression was applied to the data, the variables of older age (P = 3.93 x 10(-5)), higher BMI at baseline (P = 0.0003), and increased levels of IL-6 at year 5 (P = 0.0129) were determined to be independent predictors of the appearance of RKOA at year 10. The results were fully confirmed using longitudinal modeling of repeated measurements of the data obtained at 3 visits. The odds ratio for RKOA in subjects whose IL-6 levels were in the fourth quartile of increasing levels (versus the first quartile) was 2.74 (95% confidence interval 1.94-3.87). CONCLUSION: This followup study showed that individuals were more likely to be diagnosed as having RKOA if they had a higher BMI and increased circulating levels of IL-6. These results should stimulate more work on IL-6 as a potential therapeutic target. PMID- 19565478 TI - Spondylarthritis in HLA-B27/human beta2-microglobulin-transgenic rats is not prevented by lack of CD8. AB - OBJECTIVE: HLA-B27 predisposes to spondylarthritis by an unknown mechanism. A logical candidate mechanism is through recognition of B27 by CD8+ T cells. The purpose of this study was to examine the effects of a lack of CD8 on the spondylarthritis that develops in B27/human beta(2)-microglobulin (Hubeta(2)m) transgenic rats. METHODS: A missense mutation in the CD8a gene that causes a loss of CD8alpha expression was identified in offspring of a male Sprague-Dawley rat that had been treated with the mutagen N-ethyl-N-nitrosourea. The mutation was crossed into B27/Hubeta(2)m-transgenic lines on the Lewis background. CD8a(-/-) and CD8a(+/-) progeny were compared on a mixed SD-LEW background as well as after at least 10 backcrosses to LEW rats. CD8 function was assessed by generating cytolytic T lymphocytes (CTLs) against allogeneic DA strain antigens. RESULTS: Homozygous mutant rats showed normal CD8a and CD8b messenger RNA levels but no detectable expression of either protein and an almost complete abrogation of the allogeneic CTL response. Two disease phenotypes previously observed in different B27/Hubeta(2)m-transgenic lines also occurred in the respective CD8a(-/-) transgenic rat lines. There was no significant difference in disease prevalence or severity between CD8a(-/-) rats and CD8a(+/-) rats. CONCLUSION: All of the previously described disease manifestations in HLA-B27/Hubeta(2)m-transgenic rats arise in the absence of any functional CD8+ T cells. It thus seems unlikely that classic T cell recognition of HLA-B27 is of primary importance in this animal model. The possibility of a secondary role of a CD8-dependent mechanism cannot be entirely excluded. PMID- 19565479 TI - Enthesis inflammation in recurrent acute anterior uveitis without spondylarthritis. AB - OBJECTIVE: To investigate whether patients with idiopathic recurrent acute anterior uveitis (AAU) have enthesis alterations comparable with those in patients with spondylarthritis (SpA). METHODS: A blinded, controlled study of enthesis evident on ultrasound (US) examination was performed in 100 patients and controls classified into 5 groups, as follows: patients with confirmed SpA (group 1), patients with recurrent AAU who were positive for HLA-B27 and did not have SpA (group 2), patients with recurrent AAU who were negative for HLA-B27 and did not have SpA (group 3), patients with forms of uveitis other than those related to SpA (group 4), and healthy controls (group 5). In total, 12 enthesis locations were explored in each patient and control subject by 2 ultrasonographers who were blinded with regard to the diagnosis. A newly developed US method, the Madrid Sonography Enthesitis Index (MASEI), in which the diagnosis of SpA is determined as a cutoff score of 18 points, was used. RESULTS: A total of 1,200 entheses were explored by US in 100 patients and controls. The MASEI cutoff limit was met or exceeded by 81%, 55.6%, 40%, 10%, and 19% of the subjects in the 5 groups, respectively. The MASEI score was significantly higher in groups 1 and 2 than in groups 4 and 5. The differences between groups 1 and 3 were also found to be significant. CONCLUSION: Our findings indicate that a high percentage of HLA-B27 positive patients with idiopathic recurrent AAU without features of SpA have enthesis lesions comparable with those seen in patients with SpA. These data suggest that patients with recurrent AAU, especially those who are HLA-B27 positive, have an abortive or incomplete form of SpA. PMID- 19565480 TI - A multicenter survey of rituximab therapy for refractory antineutrophil cytoplasmic antibody-associated vasculitis. AB - OBJECTIVE: B cell depletion with rituximab has allowed remissions in relapsing or refractory antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis in small studies. The aim of this study was to determine the efficacy and safety of rituximab for ANCA-associated vasculitis in a larger multicenter cohort. This permitted comparison of rituximab dosing regimens, the value of continuing immunosuppression, and investigation of ANCA and B cell levels as re-treatment biomarkers. METHODS: Retrospective, standardized data collection from 65 sequential patients receiving rituximab for refractory ANCA-associated vasculitis at 4 centers in the UK was used. RESULTS: All patients achieved B cell depletion. Complete remission occurred in 49 of the 65 patients (75%), partial remission in 15 (23%), and no response in 1 (2%). The prednisolone dosage was reduced from 12.5 mg/day (median) to 9.0 mg/day at 6 months (P = 0.0006). Immunosuppressive therapy was withdrawn in 37 of 60 patients (62%). Twenty-eight of 49 patients who achieved full remission (57%) experienced relapse (median 11.5 months). B cell return preceded relapse in 14 of 27 patients (52%). Although ANCA levels fell after rituximab therapy, relapse was not associated with ANCA positivity or a rise in ANCA levels. Neither the initial rituximab regimen (4 infusions of 375 mg/m(2) each given 1 week apart or 2 infusions of 1 gm each given 2 weeks apart) nor withdrawal of immunosuppressive therapy (37 of 60 patients [62%]) influenced the timing of relapse. Thirty-eight patients received >or=2 courses of rituximab, and complete remission was induced or maintained in 32 of them (84%). IgM levels fell, although IgG levels remained stable. Forty-six serious adverse events occurred, including 2 episodes of late-onset neutropenia, which were attributed to rituximab. CONCLUSION: Rituximab was effective remission induction therapy for refractory ANCA-associated vasculitis in this study. There was no difference in efficacy between the 2 main treatment regimens. Continuing immunosuppression did not reduce relapses. Relapses occurred, but re-treatment was effective and safe. There was no clear influence of rituximab on the frequency of serious adverse events. ANCA and B cell levels lacked sufficient sensitivity to guide the timing of re-treatment. PMID- 19565481 TI - Fibroblast growth factor 2 is an intrinsic chondroprotective agent that suppresses ADAMTS-5 and delays cartilage degradation in murine osteoarthritis. AB - OBJECTIVE: We have previously identified in articular cartilage an abundant pool of the heparin-binding growth factor, fibroblast growth factor 2 (FGF-2), which is bound to the pericellular matrix heparan sulfate proteoglycan, perlecan. This pool of FGF-2 activates chondrocytes upon tissue loading and is released following mechanical injury. In vitro, FGF-2 suppresses interleukin-1-driven aggrecanase activity in human cartilage explants, suggesting a chondroprotective role in vivo. We undertook this study to investigate the in vivo role of FGF-2 in murine cartilage. METHODS: Basal characteristics of the articular cartilage of Fgf2(-/-) and Fgf2(+/+) mice were determined by histomorphometry, nanoindentation, and quantitative reverse transcriptase-polymerase chain reaction. The articular cartilage was graded histologically in aged mice as well as in mice in which osteoarthritis (OA) had been induced by surgical destabilization of the medial meniscus. RNA was extracted from the joints of Fgf2(-/-) and Fgf2(+/+) mice following surgery and quantitatively assessed for key regulatory molecules. The effect of subcutaneous administration of recombinant FGF-2 on OA progression was assessed in Fgf2(-/-) mice. RESULTS: Fgf2(-/-) mice were morphologically indistinguishable from wild-type (WT) animals up to age 12 weeks; the cartilage thickness and proteoglycan staining were equivalent, as was the mechanical integrity of the matrix. However, Fgf2(-/-) mice exhibited accelerated spontaneous and surgically induced OA. Surgically induced OA in Fgf2(-/-) mice was suppressed to levels in WT mice by subcutaneous administration of recombinant FGF-2. Increased disease in Fgf2(-/-) mice was associated with increased expression of messenger RNA of Adamts5, the key murine aggrecanase. CONCLUSION: These data identify FGF-2 as a novel endogenous chondroprotective agent in articular cartilage. PMID- 19565482 TI - Association of adrenergic receptor gene polymorphisms with different fibromyalgia syndrome domains. AB - OBJECTIVE: Fibromyalgia (FM) patients have signs of relentless sympathetic hyperactivity associated with hyporeactivity to stress. Adrenergic receptors (ARs) are parts of the sympathetic nervous system that are fundamental for maintenance of homeostasis. We undertook this study to correlate alpha-AR and beta-AR gene polymorphisms with the presence of FM and with different domains of the FM syndrome as measured by the Fibromyalgia Impact Questionnaire (FIQ). METHODS: We studied 78 Mexican FM patients and 48 age-matched controls as well as 78 Spanish FM patients and 71 controls. All subjects studied were women. Single nucleotide polymorphisms (SNPs) of alpha(1A)-AR (rs574584, rs1383914, rs1048101, and rs573542), beta(2)-AR (rs1042713 and rs1042714), and beta(3)-AR (rs4994) were analyzed by 5' exonuclease TaqMan polymerase chain reaction. Polymorphic haplotypes were constructed after linkage disequilibrium analysis. RESULTS: The beta(2)-AR AC haplotype was a risk factor for the presence of FM. This haplotype had an increased frequency in Mexican patients compared with Mexican controls (42.1% versus 30.5%; P = 0.04). Similarly, 50.4% of Spanish patients had this haplotype compared with 40.0% of Spanish controls (P = 0.05). In Spanish patients, the alpha(1A)-AR SNP rs1383914 was associated with the presence of FM (P = 0.01), and the alpha(1A)-AR SNP rs1048101 was linked with FIQ disability (P = 0.02). Mexican patients with the rs574584 GG genotype presented the highest FIQ score compared with Mexican patients with other genotypes (P = 0.01), and in Mexicans SNP rs574584 was associated with FIQ morning stiffness (P = 0.04) and with FIQ tiredness upon awakening (P = 0.02). CONCLUSION: AR gene polymorphisms are related to the risk of developing FM and are also linked to different domains of the FM syndrome. PMID- 19565483 TI - Pan-DR-binding Hsp60 self epitopes induce an interleukin-10-mediated immune response in rheumatoid arthritis. AB - OBJECTIVE: Human Hsp60 is expressed in the joints of patients with rheumatoid arthritis (RA) and can elicit a regulatory T cell response in the peripheral blood and synovial fluid. However, Hsp60 can also trigger strong proinflammatory pathways. Thus, to understand the nature of these Hsp60-directed responses in RA, it is necessary to study such responses at the molecular, epitope-specific level. This study was undertaken to characterize the disease specificity and function of pan-DR-binding Hsp60-derived epitopes as possible modulators of autoimmune inflammation in RA. METHODS: Lymphocyte proliferation assays (using (3)H thymidine incorporation and carboxyfluorescein diacetate succinimidyl ester [CFSE] staining) and measurement of cytokine production (using multiplex immunoassay and intracellular staining) were performed after in vitro activation of peripheral blood mononuclear cells from patients with RA, compared with healthy controls. RESULTS: A disease (RA)-specific immune recognition, characterized by T cell proliferation as well as increased production of tumor necrosis factor alpha (TNFalpha), interleukin-1beta (IL-1beta), and IL-10, was found for 3 of the 8 selected peptides in patients with RA as compared with healthy controls (P < 0.05). Intracellular cytokine staining and CFSE labeling showed that CD4+ T cells were the subset primarily responsible for both the T cell proliferation and the cytokine production in RA. Interestingly, the human peptides had a remarkably different phenotype, with a 5-10-fold higher IL 10:TNFalpha ratio, compared with that of the microbial peptides. CONCLUSION: These results suggest a disease-specific immune-modulatory role of epitope specific T cells in the inflammatory processes of RA. Therefore, these pan-DR binding epitopes could be used as a tool to study the autoreactive T cell response in RA and might be suitable candidates for use in immunotherapy. PMID- 19565484 TI - Fibroblast growth factor 2: A new key player in osteoarthritis. PMID- 19565485 TI - Intravenous immunoglobulin and salicylate differentially modulate pathogenic processes leading to vascular damage in a model of Kawasaki disease. AB - OBJECTIVE: Kawasaki disease (KD) is a multisystem vasculitis affecting children and is characterized by immune activation in the acute stage of disease. Systemic inflammation eventually subsides, although coronary arteritis persists, resulting in aneurysm formation. KD is the leading cause of acquired heart disease among children in North America. Accepted treatment guidelines include high-dose intravenous immunoglobulin (IVIG) and aspirin in the acute phase. Although this therapy is effective, the cellular and molecular mechanisms involved are not clear. The aim of this study was to examine the effect of IVIG and salicylate at each stage of disease development. METHODS: Using a murine model of KD, we established and validated several in vitro techniques to reflect 3 key steps involved in disease pathogenesis, as follows: thymidine incorporation to evaluate T cell activation, enzyme-linked immunosorbent assay to measure tumor necrosis factor alpha (TNFalpha) production, and real-time polymerase chain reaction to examine TNFalpha-mediated expression of matrix metalloproteinase 9 (MMP-9). RESULTS: At therapeutic concentrations, IVIG, but not salicylate, effectively reduced the immune response leading to TNFalpha expression. Unexpectedly, pharmacologic doses of salicylate were not able to inhibit TNFalpha production and in fact enhanced its production. Neither drug directly regulated MMP-9 expression but did so only indirectly via modulating TNFalpha. TNFalpha activity was a prerequisite for local expression of MMP-9 at the coronary artery. CONCLUSION: Therapeutic concentrations of IVIG and salicylate differentially modulate the expression of TNFalpha and its downstream effects. Further dissection of the biologic effects of aspirin in acute KD is necessary for the rational design of therapy. PMID- 19565486 TI - Changes in proximal femoral mineral geometry precede the onset of radiographic hip osteoarthritis: The study of osteoporotic fractures. AB - OBJECTIVE: Radiographic hip osteoarthritis (RHOA) is associated with increased hip areal bone mineral density (aBMD). This study was undertaken to examine whether femoral geometry is associated with RHOA independent of aBMD. METHODS: Participants in the Study of Osteoporotic Fractures in whom pelvic radiographs had been obtained at visits 1 and 5 (mean 8.3 years apart) and hip dual x-ray absorptiometry (DXA) had been performed (2 years after baseline) were included. Prevalent and incident RHOA phenotypes were defined as composite (osteophytes and joint space narrowing [JSN]), atrophic (JSN without osteophytes), or osteophytic (femoral osteophytes without JSN). Analogous definitions of progression were based on minimum joint space and total osteophyte score. Hip DXA scans were assessed using the Hip Structural Analysis program to derive geometric measures, including femoral neck length, width, and centroid position. Relative risks and 95% confidence intervals for prevalent, incident, and progressive RHOA per SD increase in geometric measure were estimated in a hip-based analysis using multinomial logistic regression with adjustment for age, body mass index, knee height, and total hip aBMD. RESULTS: In 5,245 women (mean age 72.6 years), a wider femoral neck with a more medial centroid position was associated with prevalent and incident osteophytic and composite RHOA phenotypes (P < 0.05). Increased neck width and centroid position were associated with osteophyte progression (both P < 0.05). No significant geometric associations with atrophic RHOA were found. CONCLUSION: Differences in proximal femoral bone geometry and spatial distribution of bone mass occur early in hip OA and predict prevalent, incident, and progressive osteophytic and composite phenotypes, but not the atrophic phenotype. These bone differences may reflect responses to loading occurring early in the natural history of RHOA. PMID- 19565487 TI - Variation at the ANP32A gene is associated with risk of hip osteoarthritis in women. AB - OBJECTIVE: The ANP32A gene encodes a tumor suppressor molecule that plays a regulatory role in apoptosis and interferes with canonical Wnt signaling in vitro. We undertook this study to test whether genetic variation at ANP32A was associated with osteoarthritis (OA) in women. METHODS: Single-nucleotide polymorphisms (SNPs) in the ANP32A gene were genotyped in 438 control women, 425 women with total knee replacements (TKRs), and 537 women with total hip replacements (THRs) from the Nottingham case-control study as well as in 820 women from the population-based Chingford Study cohort for whom hip and knee radiographs were available. The most highly associated SNP was further tested in women from the Rotterdam Study (131 with THRs, 633 with knee OA, and 1,567 controls) and the TwinsUK Study cohort (67 with THRs, 43 with TKRs, and 358 controls), for a total of 2,170 patients with OA and 2,849 controls. RESULTS: The ANP32A transcript was abundantly expressed in normal and OA articular cartilage. Three SNPs in the ANP32A gene were significantly associated in Nottingham patients with hip OA, but not knee OA. One of these (rs7164503) was associated with hip and knee OA in the Chingford Study cohort and with THR in the TwinsUK Study cohort, but the association was not statistically significant in the Rotterdam Study. When we combined hip data from all 4 cohorts, we found that the minor allele of rs7164503 was associated with a significantly lower risk of hip OA (Mantel-Haenszel odds ratio 0.67 [95% confidence interval 0.53-0.84], P < 3.8 x 10(-4)) and that a similar trend was observed for knee OA (Mantel-Haenszel odds ratio 0.87 [95% confidence interval 0.73-1.01], P < 0.055). CONCLUSION: Our results provide evidence suggesting that ANP32A is involved in the pathogenesis of OA of the hip. PMID- 19565488 TI - Systemic activation of the immune system induces aberrant BAFF and APRIL expression in B cells in patients with systemic lupus erythematosus. AB - OBJECTIVE: Elevated levels of BAFF and APRIL are characteristic of patients with systemic lupus erythematosus (SLE). The reasons for enhanced cytokine production are not well understood. This study was undertaken to identify the cells responsible for the overproduction of these cytokines. METHODS: BAFF expression was analyzed on peripheral blood mononuclear cells by multiparameter flow cytometry and in tissue samples by immunofluorescence staining. The levels of BAFF and APRIL mRNA were quantified in sorted B cells. In vitro cultures were used to analyze whether B cell survival and differentiation was supported by autocrine BAFF and/or APRIL. RESULTS: Aberrant activation of B cells in patients with SLE was associated with a significant up-regulation of BAFF expression in naive, memory, and plasma cells. Furthermore, strong expression of BAFF and APRIL was found in plasma cells from the lymph node, bone marrow, and kidney. The levels of BAFF and APRIL mRNA in CD19+ B cells correlated both with the titer of anti-double stranded DNA antibodies and with the SLE Disease Activity Index. In vitro experiments demonstrated that B cells released functional BAFF/APRIL upon activation. CONCLUSION: Our data show that B cells contribute to the enhanced levels of circulating BAFF and APRIL. The aberrant up-regulation of these cytokines may initiate a vicious circle in which enhanced levels of BAFF and APRIL act in an autocrine manner to reinforce the systemic activation of the humoral immune system. PMID- 19565489 TI - A new class of potent matrix metalloproteinase 13 inhibitors for potential treatment of osteoarthritis: Evidence of histologic and clinical efficacy without musculoskeletal toxicity in rat models. AB - OBJECTIVE: Matrix metalloproteinases (MMPs) have long been considered excellent targets for osteoarthritis (OA) treatment. However, clinical utility of broad spectrum MMP inhibitors developed for this purpose has been restricted by dose limiting musculoskeletal side effects observed in humans. This study was undertaken to identify a new class of potent and selective MMP-13 inhibitors that would provide histologic and clinical efficacy without musculoskeletal toxicity. METHODS: Selectivity assays were developed using catalytic domains of human MMPs. Freshly isolated bovine articular cartilage or human OA cartilage was used in in vitro cartilage degradation assays. The rat model of monoiodoacetate (MIA) induced OA was implemented for assessing the effects of MMP-13 inhibitors on cartilage degradation and joint pain. The surgical medial meniscus tear model in rats was used to evaluate the chondroprotective ability of MMP-13 inhibitors in a chronic disease model of OA. The rat model of musculoskeletal side effects (MSS) was used to assess whether selective MMP-13 inhibitors have the joint toxicity associated with broad-spectrum MMP inhibitors. RESULTS: A number of non hydroxamic acid-containing compounds that showed a high degree of potency for MMP 13 and selectivity against other MMPs were designed and synthesized. Steady-state kinetics experiments and Lineweaver-Burk plot analysis of rate versus substrate concentration with one such compound, ALS 1-0635, indicated linear, noncompetitive inhibition, and Dixon plot analysis from competition studies with a zinc chelator (acetoxyhydroxamic acid) and ALS 1-0635 demonstrated nonexclusive binding. ALS 1-0635 inhibited bovine articular cartilage degradation in a dose dependent manner (48.7% and 87.1% at 500 nM and 5,000 nM, respectively) and was effective in inhibiting interleukin-1alpha- and oncostatin M-induced C1,C2 release in human OA cartilage cultures. ALS 1-0635 modulated cartilage damage in the rat MIA model (mean +/- SEM damage score 1.3 +/- 0.3, versus 2.2 +/- 0.4 in vehicle-treated animals). Most significantly, when treated twice daily with oral ALS 1-0635, rats with surgically induced medial meniscus tear exhibited histologic evidence of chondroprotection and reduced cartilage degeneration, without observable musculoskeletal toxicity. CONCLUSION: The compounds investigated in this study represent a novel class of MMP-13 inhibitors. They are mechanistically distinct from previously reported broad-spectrum MMP inhibitors and do not exhibit the problems previously associated with these inhibitors, including selectivity, poor pharmacokinetics, and MSS liability. MMP-13 inhibitors exert chondroprotective effects and can potentially modulate joint pain, and are, therefore, uniquely suited as potential disease-modifying osteoarthritis drugs. PMID- 19565492 TI - International rheumatology networking: The 2008 American College of Rheumatology/European League Against Rheumatism exchange program. PMID- 19565491 TI - The CGGGG insertion/deletion polymorphism of the IRF5 promoter is a strong risk factor for primary Sjogren's syndrome. AB - OBJECTIVE: Interferon regulatory factor 5 is a transcription factor involved in type I interferon (IFN) secretion. This study was undertaken to investigate whether a 5-bp (CGGGG insertion/deletion) promoter polymorphism is involved in genetic predisposition to primary Sjogren's syndrome (SS) and to assess the functional consequences of this polymorphism. METHODS: The exploratory cohort consisted of 185 patients with primary SS and 157 healthy controls, and the replication cohort consisted of 200 patients with primary SS and 282 healthy controls. Levels of IRF5 messenger RNA (mRNA) were assessed at baseline and after in vitro infection with reovirus in peripheral blood mononuclear cells (PBMCs) from 30 patients with primary SS and from salivary gland epithelial cells that had been cultured for 4 weeks from patients with primary SS or sicca symptoms. RESULTS: Carriage of the IRF5 4R CGGGG allele was associated with a greatly increased risk of primary SS in both cohorts (odds ratio 2.00 [95% confidence interval 1.5-2.7], P = 6.6 x 10(-6)). The CGGGG insertion/deletion polymorphism alone was sufficient to explain the association of primary SS with IRF5. The level of IRF5 mRNA in PBMCs depended significantly on genotype (P = 0.002) and was correlated with the levels of mRNA for the IFN-induced genes MX1 and IFITM1. Cultured salivary gland epithelial cells from patients carrying the 4R CGGGG IRF5 allele showed a high level of IRF5 mRNA (P = 0.04), which was amplified after reovirus infection (P = 0.026). CONCLUSION: Our findings indicate an association of the CGGGG insertion/deletion polymorphism of the IRF5 promoter with primary SS. Patients carrying the 4R CGGGG IRF5 allele had a high level of mRNA for IRF5 in PBMCs and salivary gland epithelial cells, mainly after in vitro viral infection. Patients with high levels of mRNA for IRF5 also had high levels of mRNA for type I IFN-induced genes in PBMCs. PMID- 19565490 TI - Characterization and peripheral blood biomarker assessment of anti-Jo-1 antibody positive interstitial lung disease. AB - OBJECTIVE: Using a combination of clinical, radiographic, functional, and serum protein biomarker assessments, this study was aimed at defining the prevalence and clinical characteristics of interstitial lung disease (ILD) in a large cohort of patients with anti-Jo-1 antibodies. METHODS: A review of clinical records, pulmonary function test results, and findings on imaging studies determined the existence of ILD in anti-Jo-1 antibody-positive individuals whose data were accumulated in the University of Pittsburgh Myositis Database from 1982 to 2007. Multiplex enzyme-linked immunosorbent assays (ELISAs) for serum inflammation markers, cytokines, chemokines, and matrix metalloproteinases in different patient subgroups were performed to assess the serum proteins associated with anti-Jo-1 antibody-positive ILD. RESULTS: Among the 90 anti-Jo-1 antibody positive individuals with sufficient clinical, radiographic, and/or pulmonary function data, 77 (86%) met the criteria for ILD. While computed tomography scans revealed a variety of patterns suggestive of underlying usual interstitial pneumonia (UIP) or nonspecific interstitial pneumonia, a review of the histopathologic abnormalities in a subset of patients undergoing open lung biopsy or transplantation or whose lung tissue was obtained at autopsy (n = 22) demonstrated a preponderance of UIP and diffuse alveolar damage. Analysis by multiplex ELISA yielded statistically significant associations between anti-Jo-1 antibody-positive ILD and elevated serum levels of C-reactive protein (CRP), CXCL9, and CXCL10, which distinguished this disease entity from idiopathic pulmonary fibrosis and anti-signal recognition particle antibody-positive myositis. Recursive partitioning further demonstrated that combinations of these and other serum protein biomarkers can distinguish these disease subgroups at high levels of sensitivity and specificity. CONCLUSION: In this large cohort of anti-Jo-1 antibody-positive individuals, the incidence of ILD approached 90%. Multiplex ELISA demonstrated disease-specific associations between anti-Jo-1 antibody-positive ILD and serum levels of CRP as well as the interferon-gamma inducible chemokines CXCL9 and CXCL10, highlighting the potential of this approach to define biologically active molecules contributing to the pathogenesis of myositis-associated ILD. PMID- 19565493 TI - Adipocytokines are associated with radiographic joint damage in rheumatoid arthritis. AB - OBJECTIVE: Obesity protects against radiographic joint damage in rheumatoid arthritis (RA) through poorly defined mechanisms. Adipocytokines are produced in adipose tissue and modulate inflammatory responses and radiographic joint damage in animal models. The purpose of this study was to examine the hypothesis that adipocytokines modulate inflammation and radiographic joint damage in patients with RA. METHODS: We compared serum concentrations of leptin, resistin, adiponectin, and visfatin in 167 RA patients and 91 control subjects. The independent association between adipocytokines and body mass index (BMI), measures of inflammation (C-reactive protein [CRP], interleukin-6 [IL-6], and tumor necrosis factor alpha [TNFalpha]), and radiographic joint damage (Larsen score; n = 93 patients) was examined in RA patients by multivariable regression analysis first controlling for age, race, and sex, and then for obesity (BMI) and inflammation (TNFalpha, IL-6, and CRP). RESULTS: Concentrations of all adipocytokines were significantly higher in RA patients than in controls; for visfatin and adiponectin, this association remained significant after adjusting for BMI, inflammation, or both. Visfatin concentrations were associated with higher Larsen scores, and this association remained significant after adjustment for age, race, sex, disease duration, BMI, and inflammation (odds ratio [OR] 2.38 [95% confidence interval (95% CI) 1.32-4.29], P = 0.004). Leptin concentrations showed a positive association with the BMI (rho = 0.58, P < 0.01) and showed a negative association with the Larsen score after adjustment for inflammation (OR 0.32 [95% CI 0.17-0.61], P < 0.001), but not after adjustment for BMI (OR 0.86 [95% CI 0.42-1.73], P = 0.67). CONCLUSION: Concentrations of adipocytokines are increased in patients with RA and may modulate radiographic joint damage. Visfatin is associated with increased, and leptin with reduced, levels of radiographic joint damage. PMID- 19565494 TI - Reply to letter by Nardelli and Schell commenting on the pathogenesis of Lyme arthritis. PMID- 19565496 TI - Small ubiquitin-like modifier 1 [corrected] mediates the resistance of prosthesis loosening fibroblast-like synoviocytes against Fas-induced apoptosis. AB - OBJECTIVE: To study the expression of small ubiquitin-like modifier 1 (SUMO-1) in aseptic loosening of prosthesis implants and to investigate its role in regulating the susceptibility of prosthesis-loosening fibroblast-like synoviocytes (FLS) to Fas-induced apoptosis. METHODS: Specimens of aseptically loosened tissue were obtained at revision surgery, and the expression of SUMO-1 was analyzed by in situ hybridization. SUMO-1 levels in FLS were determined by quantitative polymerase chain reaction and Western blot analysis. Immunohistochemistry and confocal microscopy were used to study the subcellular localization of SUMO-1. The functional role of SUMO-1 in Fas-induced apoptosis of prosthesis-loosening FLS was investigated by small interfering RNA-mediated knockdown of SUMO-1 and by gene transfer of the nuclear SUMO-specific protease SENP1. RESULTS: SUMO-1 was expressed strongly in aseptically loosened tissue and was found prominently at sites adjacent to bone. Prosthesis-loosening FLS expressed levels of SUMO-1 similar to the levels expressed by rheumatoid arthritis (RA) FLS, with SUMO-1 being found mainly in promyelocytic leukemia protein nuclear bodies. Knockdown of SUMO-1 had no effect on spontaneous apoptosis but significantly increased the susceptibility of prosthesis-loosening FLS to Fas-induced apoptosis. Gene transfer of the nuclear SUMO-specific protease SENP1 reverted the apoptosis-inhibiting effects of SUMO-1. CONCLUSION: These data suggest that SUMO-1 is involved in the activation of both RA FLS and prosthesis loosening FLS by preventing these cells from undergoing apoptosis. Modification of nuclear proteins by SUMO-1 contributes to the antiapoptotic effects of SUMO-1 in prosthesis-loosening FLS, providing evidence for the specific activation of sumoylation during their differentiation. Therefore, SUMO-1 may be an interesting target for novel strategies to prevent aseptic prosthesis loosening. PMID- 19565495 TI - Risk of tuberculosis is higher with anti-tumor necrosis factor monoclonal antibody therapy than with soluble tumor necrosis factor receptor therapy: The three-year prospective French Research Axed on Tolerance of Biotherapies registry. AB - OBJECTIVE: Tuberculosis (TB) is associated with anti-tumor necrosis factor (anti TNF) monoclonal antibody (mAb) therapy, but whether this association is drug specific remains a concern. Our objective was to describe cases of TB associated with anti-TNF mAb therapy, identify risk factors, and estimate the incidence. METHODS: We conducted an incidence study and a case-control analysis to investigate the risk of newly diagnosed TB associated with the use of anti-TNF agents. As part of the French Research Axed on Tolerance of Biotherapies (RATIO) registry, for 3 years we collected cases of TB among French patients receiving anti-TNF mAb therapy for any indication; for each case, 2 patients treated with anti-TNF agents served as control subjects. RESULTS: We collected 69 cases of TB in patients treated for rheumatoid arthritis (n = 40), spondylarthritides (n = 18), inflammatory colitis (n = 9), psoriasis (n = 1) and Behcet's disease (n = 1) with infliximab (n = 36), adalimumab (n = 28), and etanercept (n = 5). None of the patients had received correct chemoprophylactic treatment. The sex- and age adjusted incidence rate of TB was 116.7 per 100,000 patient-years. The standardized incidence ratio (SIR) was 12.2 (95% confidence interval [95% CI] 9.7 15.5) and was higher for therapy with infliximab and adalimumab than for therapy with etanercept (SIR 18.6 [95% CI 13.4-25.8] and SIR 29.3 [95% CI 20.3-42.4] versus SIR 1.8 [95% CI 0.7-4.3], respectively). In the case-control analysis, exposure to infliximab or adalimumab versus etanercept was an independent risk factor for TB (odds ratio [OR] 13.3 [95% CI 2.6-69.0] and OR 17.1 [95% CI 3.6 80.6], respectively). Other risk factors were age, the first year of anti-TNF mAb treatment, and being born in an endemic area. CONCLUSION: The risk of TB is higher for patients receiving anti-TNF mAb therapy than for those receiving soluble TNF receptor therapy. The increased risk with early anti-TNF treatment and the absence of correct chemoprophylactic treatment favor the reactivation of latent TB. PMID- 19565498 TI - Functional analysis of the osteoarthritis susceptibility-associated GDF5 regulatory polymorphism. AB - OBJECTIVE: Single-nucleotide polymorphism (SNP) rs143383 (T to C) in the 5' untranslated region (5'-UTR) of GDF5 has recently been reported to be associated with osteoarthritis (OA) susceptibility, with lower expression of the risk associated T allele observed in vitro and in vivo. The in vivo studies were performed on cartilage tissue from OA patients. The present study was undertaken to expand the analysis of the effect of this SNP on GDF5 allelic expression to more joint tissue types, to investigate for cis and trans factors that interact with the SNP, and to examine novel cis-acting GDF5 regulatory polymorphisms. METHODS: Tissue samples were collected from OA patients undergoing joint replacement of the hip or knee. Nucleic acid was extracted, and, using rs143383 and an assay that discriminates and quantifies allelic expression, the relative amount of GDF5 expression from the T and C alleles was measured. Additional common variants in the GDF5 transcript sequence were interrogated as potential regulatory elements using allelic expression and luciferase reporter assays, and electrophoretic mobility shift assays were used to search for trans factors binding to rs143383. RESULTS: We observed a consistent allelic expression imbalance of GDF5 in all tissues tested, implying that the functional effect mediated by rs143383 on GDF5 expression is joint-wide. We identified a second polymorphism, located in the 3'-UTR of GDF5, that influenced allelic expression of the gene independent of rs143383. Finally, we observed differential binding of deformed epidermal autoregulatory factor 1 (DEAF-1) to the 2 alleles of rs143383. CONCLUSION: These findings show that the OA susceptibility mediated by polymorphism in GDF5 is not restricted to cartilage, emphasizing the need to consider the disease as involving the whole joint. The existence of an additional cis-acting regulatory polymorphism highlights the complexity of the regulation of expression of this important OA susceptibility locus. DEAF-1 is a trans-acting factor that merits further investigation as a potential tool for modulating GDF5 expression. PMID- 19565497 TI - Alterations of the synovial T cell repertoire in anti-citrullinated protein antibody-positive rheumatoid arthritis. AB - OBJECTIVE: The association of HLA-DRB1 alleles with anti-citrullinated protein antibodies (ACPAs) in rheumatoid arthritis (RA) suggests the potential involvement of T lymphocytes in ACPA-seropositive disease. The purpose of this study was to investigate this hypothesis by systematic histologic and molecular analyses of synovial T cells in ACPA+ versus ACPA- RA patients. METHODS: Synovial biopsy samples were obtained from 158 RA patients. Inflammation was determined histologically and immunohistochemically. RNA was extracted from peripheral blood mononuclear cells and synovial tissues obtained from 11 ACPA+ RA patients, 7 ACPA RA patients, and 10 spondylarthritis (SpA) patients (arthritis controls). T lymphocyte clonality was studied by combined quantitative and qualitative T cell receptor CDR3 length distribution (LD) analysis and direct sequencing analysis. RESULTS: ACPA+ and ACPA- RA patients were similar at both the clinical and histologic levels. At the molecular level, however, patients with ACPA+ synovitis displayed a marked elevation of qualitative CDR3 LD alterations as compared with those with ACPA- synovitis and with the SpA controls. These differences in CDR3 LD were not observed in the peripheral blood, indicating a selective recruitment and/or local expansion of T cells in the synovial compartment. The CDR3 LD alterations reflected true monoclonal or oligoclonal expansions, as confirmed by direct sequencing of the T cell receptor. The CDR3 LD alterations in RA synovium did not correlate with B cell clonal expansions but were inversely associated with synovial lymphoid neogenesis. CONCLUSION: The T cell repertoire is specifically restricted in RA patients with ACPA+ synovitis. Whereas the origin and role of these clonal alterations remain to be determined, our data suggest the preferential involvement of T lymphocytes in ACPA-seropositive RA. PMID- 19565499 TI - Severe gouty arthritis and mild neurologic symptoms due to F199C, a newly identified variant of the hypoxanthine guanine phosphoribosyltransferase. AB - A deficiency in hypoxanthine guanine phosphoribosyltransferase (HPRT) activity leads to overproduction of uric acid. According to the degree of enzymatic deficiency, a large spectrum of neurologic features can also be observed, ranging from mild or no neurologic involvement to complete Lesch-Nyhan disease. Herein, we describe a patient with hyperuricemia, juvenile-onset gouty arthritis, nephrolithiasis, and mild neurologic symptoms, attributed to a newly identified variant of the hprt gene, c.596T>G, resulting in the amino acid change p.F199C. Residual HPRT activity (8%) protected against severe neurologic involvement in this patient. Modeling of the mutated protein was used to predict the mechanisms that led to partial enzymatic activity. Careful neurologic examination is warranted in juvenile and middle-aged patients with gout, in order to detect mild symptoms that may lead to a diagnosis of HPRT deficiency. PMID- 19565500 TI - Variants in TNFAIP3, STAT4, and C12orf30 loci associated with multiple autoimmune diseases are also associated with juvenile idiopathic arthritis. AB - OBJECTIVE: Subtypes of juvenile idiopathic arthritis (JIA) share phenotypic features with other autoimmune disorders. We investigated several genetic variants associated with rheumatoid arthritis (RA) and other autoimmune disorders for association with JIA to test the hypothesis that clinically distinct phenotypes share common genetic susceptibility factors. METHODS: Cases were 445 children with JIA, and controls were 643 healthy adults. Using the TaqMan assay, subjects were genotyped for 8 single-nucleotide polymorphisms in 7 loci including rs10499194 and rs6920220 in the TNFAIP3 locus, rs6679677 in the RSBN1 locus, rs17696736 in the C12orf30 locus, rs3761847 in the TRAF1/C5 locus, rs2104286 in the IL2RA locus, rs7574865 in the STAT4 locus, and rs2542151 in the PTPN2 locus. Alleles and genotypes were analyzed for association with JIA and JIA subtypes. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. RESULTS: The strongest associations with JIA risk or protection were observed for TNFAIP3 variants rs10499194 (OR 0.74 [95% CI 0.61-0.91], P < 0.004) and rs6920220 (OR 1.30 [95% CI 1.05-1.61], P = 0.015). We also observed associations between JIA and both STAT4 (OR 1.24 [95% CI 1.02-1.51], P = 0.029) and C12orf30 (OR 1.20 [95% CI 1.01-1.43], P = 0.041) variants. The PTPN2 variant rs2542151 deviated from Hardy-Weinberg equilibrium and was excluded from analyses. Variants in IL2RA, TRAF1/C5, and RSBN1 were not associated with JIA. After stratification by JIA subtype, the TNFAIP3 and C12orf30 variants were associated with oligoarticular JIA, while the STAT4 variant was associated primarily with polyarticular JIA. CONCLUSION: We have demonstrated associations between JIA and variants in the TNFAIP3, STAT4, and C12orf30 regions that have previously shown associations with other autoimmune diseases, including RA and systemic lupus erythematosus. Our results suggest that clinically distinct autoimmune phenotypes share common genetic susceptibility factors. PMID- 19565501 TI - Transgenic disruption of glucocorticoid signaling in mature osteoblasts and osteocytes attenuates K/BxN mouse serum-induced arthritis in vivo. AB - OBJECTIVE: Endogenous glucocorticoids (GCs) modulate numerous biologic systems involved in the initiation and maintenance of arthritis. Bone cells play a critical role in the progression of arthritis, and some of the effects of GCs on inflammation may be mediated via these cells. The aim of this study was to investigate the impact of osteoblast-targeted disruption of GC signaling on joint inflammation, cartilage damage, and bone metabolism in the K/BxN mouse serum transfer model of autoimmune arthritis. METHODS: Intracellular GC signaling was disrupted in osteoblasts through transgenic overexpression of 11beta hydroxysteroid dehydrogenase type 2 under the control of a type I collagen promoter. Arthritis was induced in 5-week-old male transgenic mice and their wild type (WT) littermates, and paw swelling was assessed daily until the mice were killed. The mice were examined by histology, histomorphometry, and microfocal computed tomography, and serum was analyzed for cytokines, adrenocorticotropic hormone, and corticosterone. RESULTS: Acute arthritis developed in both transgenic and WT mice treated with K/BxN mouse serum. However, the arthritis and local inflammatory activity were significantly attenuated in transgenic mice, as judged by clinical and histologic indices of inflammation and cartilage damage. Bone turnover and bone volume remained unchanged in arthritic transgenic mice, while WT mice exhibited stimulated bone resorption, suppressed osteoblast activity, and significantly reduced bone volume, compatible with the known effects of active inflammation on bone. Circulating levels of proinflammatory cytokines tended to be lower in arthritic transgenic mice than in control transgenic mice. CONCLUSION: Disruption of GC signaling in osteoblasts significantly attenuates K/BxN mouse serum-induced autoimmune arthritis in mice. These data suggest that osteoblasts modulate the immune-mediated inflammatory response via a GC-dependent pathway. PMID- 19565502 TI - Challenges in mapping non-HLA-DRB1 major histocompatibility genes in rheumatoid arthritis: comment on the article by Vignal et al. PMID- 19565503 TI - Role of osteopontin in induction of monocyte chemoattractant protein 1 and macrophage inflammatory protein 1beta through the NF-kappaB and MAPK pathways in rheumatoid arthritis. AB - OBJECTIVE: Osteopontin (OPN) is a proinflammatory protein with a critical role in leukocyte migration. Although OPN has been implicated in rheumatoid arthritis (RA), its underlying mechanism remains unknown. In this study, we investigated the role and molecular mechanism of OPN in the induction of 2 key chemokines, monocyte chemoattractant protein 1 (MCP-1) and macrophage inflammatory protein 1beta (MIP-1beta), in RA. METHODS: Enzyme-linked immunosorbent assay and quantitative polymerase chain reaction were used to determine chemokine expression. Leukocyte migration in the presence of OPN was measured by chemotaxis assay. Signaling and molecular events were analyzed by immunoblotting and chromatin immunoprecipitation. RESULTS: The effect of OPN on inflammatory cell migration was mediated through its unique property of inducing the expression of MCP-1 and MIP-1beta in CD14+ monocytes. The concentration of OPN was significantly elevated in RA patients and appeared to correlate with the serum levels of inflammation markers and increased expression of MCP-1 or MIP-1beta in monocytes in RA patients. Endogenous production of OPN in RA synovial fluid was attributable to increased production of MCP-1 or MIP-1beta, and this effect could be blocked by an anti-OPN antibody. Furthermore, the structural motif responsible for this property resided within residues 50-83 of human OPN, sparing the known RGD or SVVYGLR sequences. It was evident that the effect of OPN on chemokine expression was mediated through both the NF-kappaB and MAPK pathways, involving the activation of IKKbeta, p38, and JNK. CONCLUSION: These results support a unique role of OPN in leukocyte migration, in the context of perpetuation of rheumatoid synovitis through the induction of MCP-1 and MIP-1beta. PMID- 19565504 TI - Gene expression signatures in polyarticular juvenile idiopathic arthritis demonstrate disease heterogeneity and offer a molecular classification of disease subsets. AB - OBJECTIVE: To determine whether peripheral blood mononuclear cells (PBMCs) from children with recent-onset polyarticular juvenile idiopathic arthritis (JIA) exhibit biologically or clinically informative gene expression signatures. METHODS: Peripheral blood samples were obtained from 59 healthy children and 61 children with polyarticular JIA prior to treatment with second-line medications, such as methotrexate or biologic agents. RNA was extracted from isolated mononuclear cells, fluorescence labeled, and hybridized to commercial gene expression microarrays (Affymetrix HG-U133 Plus 2.0). Data were analyzed using analysis of variance at a 5% false discovery rate threshold after robust multichip analysis preprocessing and distance-weighted discrimination normalization. RESULTS: Initial analysis revealed 873 probe sets for genes that were differentially expressed between polyarticular JIA patients and healthy controls. Hierarchical clustering of these probe sets distinguished 3 subgroups within the polyarticular JIA group. Prototypical patients within each subgroup were identified and used to define subgroup-specific gene expression signatures. One of these signatures was associated with monocyte markers, another with transforming growth factor beta-inducible genes, and a third with immediate early genes. Correlation of gene expression signatures with clinical and biologic features of JIA subgroups suggested relevance to aspects of disease activity and supported the division of polyarticular JIA into distinct subsets. CONCLUSION: Gene expression signatures in PBMCs from patients with recent-onset polyarticular JIA reflect discrete disease processes and offer a molecular classification of disease. PMID- 19565505 TI - Pivotal role of connective tissue growth factor in lung fibrosis: MAPK-dependent transcriptional activation of type I collagen. AB - OBJECTIVE: Connective tissue growth factor (CTGF; CCN2) is overexpressed in systemic sclerosis (SSc) and has been hypothesized to be a key mediator of the pulmonary fibrosis frequently observed in this disease. CTGF is induced by transforming growth factor beta (TGFbeta) and is a mediator of some profibrotic effects of TGFbeta in vitro. This study was undertaken to investigate the role of CTGF in enhanced expression of type I collagen in bleomycin-induced lung fibrosis, and to delineate the mechanisms of action underlying the effects of CTGF on Col1a2 (collagen gene type I alpha2) in this mouse model and in human pulmonary fibroblasts. METHODS: Transgenic mice that were carrying luciferase and beta-galactosidase reporter genes driven by the Col1a2 enhancer/promoter and the CTGF promoter, respectively, were injected with bleomycin to induce lung fibrosis (or saline as control), and the extracted pulmonary fibroblasts were incubated with CTGF blocking agents. In vitro, transient transfection, promoter/reporter constructs, and electrophoretic mobility shift assays were used to determine the mechanisms of action of CTGF in pulmonary fibroblasts. RESULTS: In the mouse lung tissue, CTGF expression and promoter activity peaked 1 week after bleomycin challenge, whereas type I collagen expression and Col1a2 promoter activity peaked 2 weeks postchallenge. Fibroblasts isolated from the mouse lungs 14 days after bleomycin treatment retained a profibrotic expression pattern, characterized by greatly elevated levels of type I collagen and CTGF protein and increased promoter activity. In vitro, inhibition of CTGF by specific small interfering RNA and neutralizing antibodies reduced the collagen protein expression and Col1a2 promoter activity. Moreover, in vivo, anti-CTGF antibodies applied after bleomycin challenge significantly reduced the Col1a2 promoter activity by approximately 25%. The enhanced Col1a2 promoter activity in fibroblasts from bleomycin-treated lungs was partly dependent on Smad signaling, whereas CTGF acted on the Col1a2 promoter by a mechanism that was independent of the Smad binding site, but was, instead, dependent on the ERK-1/2 and JNK MAPK pathways. The CTGF effect was mapped to the proximal promoter region surrounding the inverted CCAAT box, possibly involving CREB and c-Jun. In human lung fibroblasts, the human COL1A2 promoter responded in a similar manner, and the mechanisms of action also involved ERK-1/2 and JNK signaling. CONCLUSION: Our results clearly define a direct profibrotic effect of CTGF and demonstrate its contribution to lung fibrosis through transcriptional activation of Col1a2. Blocking strategies revealed the signaling mechanisms involved. These findings show CTGF to be a rational target for therapy in fibrotic diseases such as SSc. PMID- 19565506 TI - RNA helicase encoded by melanoma differentiation-associated gene 5 is a major autoantigen in patients with clinically amyopathic dermatomyositis: Association with rapidly progressive interstitial lung disease. AB - OBJECTIVE: To identify the autoantigen recognized by the autoantibody that is associated with clinically amyopathic dermatomyositis (C-ADM) and rapidly progressive interstitial lung disease (ILD). METHODS: An anti-CADM-140 antibody positive prototype serum sample was used to screen a HeLa cell-derived complementary DNA (cDNA) library. Selected cDNA clones were further evaluated by immunoprecipitation of their in vitro-transcribed and in vitro-translated products using anti-CADM-140 antibody-positive and anti-CADM-140 antibody negative sera. The lysates of COS-7 cells transfected with the putative antigen were similarly tested. An enzyme-linked immunosorbent assay (ELISA) to detect the anti-CADM-140 antibody was established using a recombinant CADM-140 antigen, and its specificity and sensitivity for C-ADM and rapidly progressive ILD were assessed in 294 patients with various connective tissue diseases. RESULTS: By cDNA library screening and immunoprecipitation of in vitro-transcribed and in vitro-translated products, we obtained a cDNA clone encoding melanoma differentiation-associated gene 5 (MDA-5). The anti-CADM-140 antibodies in patients' sera specifically reacted with MDA-5 protein expressed in cells transfected with full-length MDA-5 cDNA, confirming the identity of MDA-5 as the CADM-140 autoantigen. The ELISA, using recombinant MDA-5 protein as the antigen, showed an analytical sensitivity of 85% and analytical specificity of 100%, in comparison with the "gold standard" immunoprecipitation assay, and was useful for identifying patients with C-ADM and/or rapidly progressive ILD. CONCLUSION: Given that RNA helicase encoded by MDA-5 is a critical molecule involved in the innate immune defense against viruses, viral infection may play an important role in the pathogenesis of C-ADM and rapidly progressive ILD. Moreover, our ELISA using recombinant MDA-5 protein makes detection of the anti-CADM-140 antibody routinely available. PMID- 19565507 TI - Anti-cyclic citrullinated peptide antibodies from rheumatoid arthritis patients activate complement via both the classical and alternative pathways. AB - OBJECTIVE: It has been suggested that anti-citrullinated protein antibodies (ACPAs) play an important role in the pathogenesis of rheumatoid arthritis (RA). To exert their pathologic effects, ACPAs must recruit immune effector mechanisms such as activation of the complement system. Mouse models of RA have shown that, surprisingly, arthritogenic antibodies activate the alternative pathway of complement rather than the expected classical pathway. This study was undertaken to investigate whether human anti-cyclic citrullinated peptide (anti-CCP) antibodies activate the complement system in vitro and, if so, which pathways of complement activation are used. METHODS: We set up novel assays to analyze complement activation by anti-CCP antibodies, using cyclic citrullinated peptide coated plates, specific buffers, and normal and complement-deficient sera as a source of complement. RESULTS: Anti-CCP antibodies activated complement in a dose dependent manner via the classical pathway of complement, and, surprisingly, via the alternative pathway of complement. The lectin pathway was not activated by anti-CCP antibodies. Complement activation proceeded in vitro up to the formation of the membrane attack complex, indicating that all activation steps, including the release of C5a, took place. CONCLUSION: Our findings indicate that anti-CCP antibodies activate the complement system in vitro via the classical and alternative pathways but not via the lectin pathway. These findings are relevant for the design of interventions aimed at inhibition of complement-mediated damage in RA. PMID- 19565508 TI - The clinical relevance of a prediction rule for disease outcome in patients with undifferentiated arthritis: comment on the article by van der Helm-van Mil et al. PMID- 19565509 TI - B lymphocyte cytokines and rheumatic autoimmune disease. PMID- 19565510 TI - Mortality in rheumatoid arthritis patients with heart failure: comment on the article by Davis et al. PMID- 19565512 TI - Disease remission state in patients treated with the combination of tumor necrosis factor blockade and methotrexate or with disease-modifying antirheumatic drugs: A clinical and imaging comparative study. AB - OBJECTIVE: For patients with rheumatoid arthritis (RA) in remission who are receiving disease-modifying antirheumatic drugs (DMARDs), radiographic progression correlates with imaging-detected synovitis as measured by power Doppler activity. In contrast, patients with disease in remission who are receiving the combination of tumor necrosis factor (TNF) blockade with methotrexate (MTX) (combination treatment) have reduced radiographic damage for the equivalent clinical state. We undertook this study to determine whether the difference in radiographic outcome is a result of more complete suppression of imaging-detected synovitis. METHODS: One hundred patients with RA in remission (Disease Activity Score in 28 joints [DAS28] <2.6) for at least 6 months while receiving either combination treatment (n = 50) or DMARDs (n = 50) were matched for clinical variables. Ultrasound of metacarpophalangeal joints 1-5 and the wrist joints was performed. Remission according to imaging results was defined as a score of 0 for both grey scale synovitis and power Doppler activity. RESULTS: In patients receiving combination treatment or DMARDs (median DAS28 1.65 versus 1.78, median disease duration 120 months versus 90 months, and median duration of remission 13 months versus 18 months), the proportion with remission according to imaging results was not significantly different (10% versus 16%, respectively). The combination treatment group had more grey scale synovitis (P < 0.001) but similar power Doppler activity (48% versus 60%, respectively; P = 0.229) in any joint as compared with the DMARD group. Results were not affected by stratification for duration of disease or remission. CONCLUSION: In RA patients with disease in remission, imaging-detected synovitis persists, with power Doppler activity seen in >or=48% of the patients regardless of therapy. These results suggest that superior radiographic outcomes in patients treated with the combination of TNF blockade and MTX may not be due to complete suppression of imaging-detected synovitis. PMID- 19565514 TI - Induction of CCL13 expression in synovial fibroblasts highlights a significant role of oncostatin M in rheumatoid arthritis. AB - OBJECTIVE: To investigate the molecular mechanisms of CCL13/monocyte chemoattractant protein 4 (MCP-4) chemokine expression through proinflammatory cytokines in different primary human fibroblasts and the contribution of CCL13 to monocyte migration. METHODS: Using RNase protection assays and enzyme-linked immunosorbent assays, we quantified the expression of CCL13 compared with that of CCL2/MCP-1 in primary human fibroblasts. Boyden chamber assays were performed to determine the importance of CCL13 for migration of primary monocytes. Pharmacologic inhibitors as well as small interfering RNA knockdown approaches were used to investigate the signaling pathways regulating CCL13 expression. RESULTS: The interleukin-6 (IL-6)-type cytokine oncostatin M (OSM) was a powerful inducer of CCL13 expression in primary synovial fibroblasts from patients with rheumatoid arthritis (RA) as well as those from healthy control subjects but not in other types of fibroblasts. Neither IL-6 nor tumor necrosis factor alpha could stimulate the expression of CCL13 in synovial fibroblasts; IL-1beta was a very weak inducer. Synovial fibroblasts from patients with RA constitutively produced low amounts of CCL13, which was partially dependent on constitutive production of OSM. By investigating the underlying molecular mechanism, we identified STAT-5, ERK-1/2, and p38 as critical factors involved in OSM-dependent transcription and messenger RNA stabilization of CCL13. CONCLUSION: In contrast to other prominent cytokines involved in the pathogenesis of RA, OSM can strongly up-regulate the expression of CCL13, a chemokine recently identified in the synovial fluid of patients with RA. Despite potent OSM-induced signal transduction in all types of fibroblasts analyzed, only synovial fibroblasts secreted CCL13, which might be indicative of tissue-specific imprinting of different fibroblasts during development. PMID- 19565515 TI - Clinical image: Cerebral amyloid angiopathy. PMID- 19565513 TI - Subtype-specific peripheral blood gene expression profiles in recent-onset juvenile idiopathic arthritis. AB - OBJECTIVE: To identify differences in peripheral blood gene expression between patients with different subclasses of juvenile idiopathic arthritis (JIA) and healthy controls in a multicenter study of patients with recent-onset JIA prior to treatment with disease-modifying antirheumatic drugs (DMARDs) or biologic agents. METHODS: Peripheral blood mononuclear cells (PBMCs) from 59 healthy children and 136 patients with JIA (28 with enthesitis-related arthritis [ERA], 42 with persistent oligoarthritis, 45 with rheumatoid factor [RF]-negative polyarthritis, and 21 with systemic disease) were isolated from whole blood. Poly(A) RNA was labeled using a commercial RNA amplification and labeling system (NuGEN Ovation), and gene expression profiles were obtained using commercial expression microarrays (Affymetrix HG-U133 Plus 2.0). RESULTS: A total of 9,501 differentially expressed probe sets were identified among the JIA subtypes and controls (by analysis of variance; false discovery rate 5%). Specifically, 193, 1,036, 873, and 7,595 probe sets were different in PBMCs from the controls compared with those from the ERA, persistent oligoarthritis, RF-negative polyarthritis, and systemic JIA patients, respectively. In patients with persistent oligoarthritis, RF-negative polyarthritis, and systemic JIA subtypes, up-regulation of genes associated with interleukin-10 (IL-10) signaling was prominent. A hemoglobin cluster was identified that was underexpressed in ERA patients but overexpressed in systemic JIA patients. The influence of JAK/STAT, ERK/MAPK, IL-2, and B cell receptor signaling pathways was evident in patients with persistent oligoarthritis. In systemic JIA, up-regulation of innate immune pathways, including IL-6, Toll-like receptor/IL-1 receptor, and peroxisome proliferator-activated receptor signaling, were noted, along with down-regulation of gene networks related to natural killer cells and T cells. Complement and coagulation pathways were up-regulated in systemic JIA, with a subset of these genes being differentially expressed in other subtypes as well. CONCLUSION: Expression analysis identified differentially expressed genes in PBMCs obtained early in the disease from patients with different subtypes of JIA and in healthy controls, providing evidence of immunobiologic differences between these forms of childhood arthritis. PMID- 19565516 TI - Relationship between elevated cerebrospinal fluid levels of plasminogen activator inhibitor 1 and neuronal destruction in patients with neuropsychiatric systemic lupus erythematosus. AB - OBJECTIVE: A homeostatic imbalance between coagulation and fibrinolysis might occur intrathecally in neuropsychiatric systemic lupus erythematosus (NPSLE). However, there are no published data on levels of fibrinolytic factors in the cerebrospinal fluid (CSF) of patients with NPSLE. The present study was undertaken to assess CSF levels of fibrinolytic molecules, including urokinase plasminogen activator (uPA), tissue plasminogen activator (tPA), D-dimer, and plasminogen activator inhibitor 1 (PAI-1), in SLE patients with clinically verified neuropsychiatric involvement and to compare these levels with those in SLE patients without neuropsychiatric involvement and in healthy subjects. METHODS: Levels of uPA, tPA, and PAI-1 were assessed in CSF from 94 patients with SLE (33 who had NPSLE, 56 who did not have NPSLE, and 5 who were positive for antiphospholipid antibody [not included in the NPSLE or non-NPSLE group]) and from 53 age-matched controls. Patients were evaluated clinically, with magnetic resonance imaging of the brain, analyses of neuronal/glial degradation products in CSF, and neuropsychiatric testing. RESULTS: In the group of patients with NPSLE, intrathecal PAI-1 levels were significantly elevated compared with levels in SLE patients without overt neuropsychiatric involvement (P < 0.05) and in healthy controls (P < 0.001). In contrast, intrathecal levels of uPA did not differ significantly. Intrathecal levels of PAI-1 correlated significantly with CSF levels of interleukin-6 (IL-6) (r = 0.34, P < 0.001) and IL-8 (r = 0.33, P < 0.001). Importantly, increased PAI-1 and D-dimer levels were observed in SLE patients who had pathologically elevated levels of glial fibrillary acidic protein, neurofilament triplet protein, and tau protein in CSF. CONCLUSION: Intrathecal release of PAI-1 is increased in patients with NPSLE. This results in impaired fibrinolysis, which might contribute to neuronal and astrocytic damage in NPSLE. PMID- 19565522 TI - Analysis of Borrelia burgdorferi genotypes in patients with Lyme arthritis: High frequency of ribosomal RNA intergenic spacer type 1 strains in antibiotic refractory arthritis. AB - OBJECTIVE: Most of the Borrelia burgdorferi genotypes have been isolated from erythema migrans (EM) skin lesions in patients with Lyme disease. OspC type K strains, which are 16S-23S ribosomal RNA intergenic spacer type 2 (RST2) strains, are most commonly recovered, but a higher percentage of OspC type A strains (RST1), the next most commonly recovered type, is detectable in blood. The goal of this study was to determine the B burgdorferi genotypes in the joints of patients with Lyme arthritis. METHODS: Joint fluid samples from 124 patients seen over a 30-year period were analyzed for OspC types by semi-nested polymerase chain reaction (PCR) and sequencing, and for RSTs by nested PCR and restriction fragment length polymorphism analysis. These results were correlated with clinical outcome. RESULTS: OspC and RST genotypes were identified in 49 of the 124 joint fluid samples (40%). In these 49 samples, OspC type K strains (RST2) were identified in 21 samples (43%), OspC type A strains (RST1) were identified in 11 samples (22%), and 8 other OspC types and all 3 RSTs were identified among the remaining 17 samples (35%). However, among the 17 patients who had been treated with antibiotics according to current guidelines, all 7 patients who were infected with RST1 strains had antibiotic-refractory arthritis, compared with 4 of 6 patients infected with RST2 strains and only 1 of 4 infected with RST3 strains (P = 0.03). CONCLUSION: Most of the B burgdorferi genotypes, particularly OspC type K (RST2), were identified in the joint fluid of patients with Lyme arthritis, and the genotype frequencies found in joints reflected those in EM skin lesions. However, RST1 strains were most frequent in patients with antibiotic-refractory arthritis. Our results help to further the understanding of the differential pathogenicity of strains of B burgdorferi. PMID- 19565523 TI - Grueneberg ganglion olfactory subsystem employs a cGMP signaling pathway. AB - The mammalian olfactory sense employs several olfactory subsystems situated at characteristic locations in the nasal cavity to detect and report on different classes of odors. These olfactory subsystems use different neuronal signal transduction pathways, receptor expression repertoires, and axonal projection targets. The Grueneberg ganglion (GG) is a newly appreciated olfactory subsystem with receptor neurons located just inside of the nostrils that project axons to a unique domain of interconnected glomeruli in the caudal olfactory bulb. It is not well understood how the GG relates to other olfactory subsystems in contributing to the olfactory sense. Furthermore, the range of chemoreceptors and the signal transduction cascade utilized by the GG have remained mysterious. To resolve these unknowns, we explored the molecular relationship between the GG and the GC D neurons, another olfactory subsystem that innervates similarly interconnected glomeruli in the same bulbar region. We found that mouse GG neurons express the cGMP-associated signaling proteins phosphodiesterase 2a, cGMP-dependent kinase II, and cyclic nucleotide gated channel subunit A3 coupled to a chemoreceptor repertoire of cilia-localized particulate guanylyl cyclases (pGC-G and pGC-A). The primary cGMP signaling pathway of the GG is shared with the GC-D neurons, unifying their target glomeruli as a unique center of olfactory cGMP signal transduction. However, the distinct chemoreceptor repertoire in the GG suggests that the GG is an independent olfactory subsystem. This subsystem is well suited to detect a unique set of odors and to mediate behaviors that remained intact in previous olfactory perturbations. PMID- 19565525 TI - The endometrial-myometrial junction: a fresh look at a busy crossing. PMID- 19565526 TI - Three-dimensional sonographic imaging of fetal bilateral cleft lip and palate in the first trimester. PMID- 19565527 TI - Biometry of the pubovisceral muscle and levator hiatus assessed by three dimensional ultrasound in females with bladder exstrophy-epispadias complex after functional reconstruction. AB - OBJECTIVE: The congenital bony and musculoskeletal defect of the pelvis in bladder exstrophy-epispadias complex (BEEC) highly predisposes females to uterine prolapse. There is a paucity of knowledge on the anatomy of the pelvic soft tissue structures following surgery. The aim of this study was to investigate with transperineal three-dimensional (3D) ultrasound the pubovisceral muscle in females with BEEC who had undergone surgical reconstruction in childhood. METHODS: In a cross-sectional observational study we examined 12 Caucasian female BEEC patients, mean age 19.9 (range, 15.5-27.4) years, from a single center after a single-stage functional reconstruction with closure of the anterior pelvic ring. As a control group we used 13 Caucasian nulligravidae. 3D transperineal ultrasound volumes were acquired with the patient at rest in the supine position and with an empty bladder, and established pelvic floor parameters were measured. Analysis was conducted offline by two independent investigators. RESULTS: No statistical difference between the BEEC patients and the control group was observed in the anteroposterior diameter or the area of the levator hiatus, or in the maximal thickness of the levator muscle. However, significantly greater values were observed in BEEC patients in the transverse diameter of the levator hiatus (mean, 4.31 vs. 3.81 cm, P = 0.046) and in the levator angle (mean, 80.1 vs. 70.0 degrees, P = 0.040). The measurements obtained in the control group were consistent with those previously reported in the literature. CONCLUSIONS: This is the first study showing that transperineal 3D ultrasound can be used for the assessment of BEEC patients after functional reconstruction. Biometric pelvic floor parameters may be useful in the long-term follow-up of BEEC patients. PMID- 19565524 TI - Approaches to measuring the effects of wake-promoting drugs: a focus on cognitive function. AB - OBJECTIVES: In clinical drug development, wakefulness and wake-promotion may be assessed by a large number of scales and questionnaires. Objective assessment of wakefulness is most commonly made using sleep latency/maintenance of wakefulness tests, polysomnography and/or behavioral measures. The purpose of the present review is to highlight the degree of overlap in the assessment of wakefulness and cognition, with consideration of assessment techniques and the underlying neurobiology of both concepts. DESIGN: Reviews of four key areas were conducted: commonly used techniques in the assessment of wakefulness; neurobiology of sleep/wake and cognition; targets of wake promoting and/or cognition enhancing drugs; and ongoing clinical trials investigating wake promoting effects. RESULTS: There is clear overlap between the assessment of wakefulness and cognition. There are common techniques which may be used to assess both concepts; aspects of the neurobiology of both concepts may be closely related; and wake-promoting drugs may have nootropic properties (and vice versa). Clinical trials of wake-promoting drugs often, though not routinely, assess aspects of cognition. CONCLUSIONS: Routine and broad assessment of cognition in the development of wake-promoting drugs may reveal important nootropic effects, which are not secondary to alertness/wakefulness, whilst existing cognitive enhancers may have underexplored or unknown wake promoting properties. PMID- 19565529 TI - Predisposing factors for massive hemorrhage during Cesarean section in patients with placenta previa. AB - OBJECTIVES: To investigate whether maternal history and ultrasound findings can be predictors for massive hemorrhage during Cesarean section in patients with placenta previa and adherence of the placenta. METHODS: We reviewed 127 singleton pregnancies with placenta previa. Maternal history, antenatal ultrasound findings of the placenta, including location, presence of placental lacunae, lack of a clear zone, presence of sponge-like findings of the cervix and presence of a marginal sinus in cases of placenta previa were reviewed retrospectively, and their association with amount of bleeding during Cesarean section was analyzed. RESULTS: Logistic regression analysis revealed that advanced maternal age (odds ratio (OR), 5.4; 95% CI, 1.8-16.4), previous Cesarean section (OR, 20.4; 95% CI, 4.0-105.2) and sponge-like findings in the cervix (OR, 5.6; 95% CI, 1.8-17.0) were associated with massive bleeding (> 2500 mL). Placental adherence occurred in five cases and was more frequent in cases where the placenta was located at the site of the scar of a previous Cesarean section (OR, 123.1; 95% CI, 4.5 3395.2) and where there was lack of a clear zone (OR, 48.0; 95% CI, 3.8-604.7). CONCLUSIONS: Advanced maternal age, previous Cesarean section and presence of sponge-like findings in the cervix are risk factors for massive bleeding during Cesarean section in cases of placenta previa, regardless of whether placental adherence is present. Placental location on the scar of a previous Cesarean section and lack of a clear zone are risk factors for placental adherence. When these findings are identified preoperatively, management should be tailored accordingly. PMID- 19565528 TI - Myocardial tissue Doppler assessment of diastolic function in the growth restricted fetus. AB - OBJECTIVES: Myocardial tissue Doppler (MTD) is a technique for the measurement of myocardial velocities in systole and diastole. This study evaluates the use of MTD in the assessment of cardiac diastolic function in fetuses with intrauterine growth restriction (IUGR) with placental insufficiency and in appropriate-for gestational age (AGA) fetuses with hypertensive mothers. METHODS: This study included 14 IUGR fetuses with placental insufficiency (Group 1), 13 AGA fetuses with hypertensive mothers (Group 2), and 29 AGA fetuses with healthy mothers (Group 3). Patients with other diseases were excluded. All fetuses had a gestational age of at least 25 weeks. MTD assessment was performed with the sample volume placed at the basal segment of the left ventricular side wall (LV), the interventricular septum (IVS) and the right ventricular free wall (RV). E'/A' ratios were calculated for each location. Mitral and tricuspid flows were assessed by conventional spectral pulsed Doppler. RESULTS: Significant differences were found between groups for E'/A' at the IVS (P < 0.001) and LV (P = 0.009), with a higher mean (+/- SD) value in Group 1 (IVS: 0.92 +/- 0.28; LV: 0.85 +/- 0.19) than in Groups 2 (IVS: 0.62 +/- 0.09; LV: 0.68 +/- 0.14) and 3 (IVS: 0.71 +/- 0.14; LV: 0.69 +/- 0.15). In the RV, there was no statistically significant difference in the E'/A' ratio (P = 0.2). No differences were observed for mitral and tricuspid flow velocities and ratios. CONCLUSIONS: Using MTD, the ratios between early and late diastolic myocardial velocities are higher in IUGR fetuses than in AGA fetuses at the IVS and the LV, regardless of the presence of maternal hypertension. MTD may be more sensitive than atrioventricular spectral Doppler for the detection of ventricular diastolic dysfunction in IUGR fetuses. PMID- 19565530 TI - Recurring eczema during pregnancy and after delivery due to sensitization caused by an ultrasound gel. PMID- 19565531 TI - Who should be allowed to perform amniocentesis and chorionic villus sampling? PMID- 19565532 TI - Three-dimensional ultrasound detection of abnormally located intrauterine contraceptive devices which are a source of pelvic pain and abnormal bleeding. AB - OBJECTIVE: To determine whether intrauterine contraceptive devices (IUDs) that are located abnormally within the myometrium or cervix cause a higher incidence of pelvic pain and abnormal bleeding compared with normally positioned devices. METHODS: Over a period of 9 months, all patients with an IUD presenting at our unit for two-dimensional pelvic ultrasound underwent a three-dimensional (3D) volume reconstruction of the coronal view, to visualize the entire IUD within the cavity. The IUD was deemed malpositioned if any part extended past the cavity, into the myometrium or cervix. The indications for ultrasound were recorded at presentation for the exam. The presenting symptoms of patients with an abnormally located IUD were compared with those with normally positioned ones. RESULTS: Among 167 consecutive patients with an IUD evaluated using the 3D reconstructed coronal view, 28 (16.8%) had an IUD with side arms abnormally located within the myometrium. The abnormal positioning of the IUD arms was only detected using the 3D coronal view. A higher proportion of patients with an abnormally located IUD presented with bleeding (35.7%) or pain (39.3%) compared with those with normally positioned IUDs (15.1% with bleeding and 19.4% with pain) (P = 0.02 and 0.03, respectively). Seventy-five percent of patients with an abnormally located IUD presented with bleeding or pain compared with 34.5% of those whose IUD was normally placed (P = 0.0001). Twenty of 21 patients with an abnormally located IUD presenting with pelvic pain or bleeding reported improvement in their symptoms after IUD removal. CONCLUSION: A 3D coronal view of the uterus is useful in the visualization of IUDs. The coronal view showing the entire device and its position within the uterus may help in identifying the cause of pelvic pain and bleeding in patients with an embedded IUD. PMID- 19565533 TI - Cavum veli interpositi cyst: prenatal diagnosis and postnatal outcome. AB - OBJECTIVE: The cavum veli interpositi (CVI) is a space within the double-layered tela choroidea of the third ventricle. Occasionally, this space is fluid-filled and sonographically visible as an interhemispheric anechoic cyst. Because of its rarity, the incidence of CVI cyst is undetermined and the outcome of affected individuals has been found to be variable. The aim of this study was to report our experience of the sonographic findings and outcome of fetuses affected by CVI cysts. METHODS: In five fetuses with a CVI cyst, we performed targeted prenatal ultrasound scans of intracranial structures and a detailed anatomical survey to rule out associated malformations. Follow-up consisted of neurological examination and neurosonography. RESULTS: The CVI cyst appeared as a well-defined anechoic lesion without adjacent mass effect. In all fetuses the cyst was single and in two cases it enlarged slightly during pregnancy. The cyst was isolated in three fetuses and associated with borderline ventriculomegaly in two. A single umbilical artery was the only associated extracranial anomaly and this was detected in only one fetus. Neurosonography confirmed the presence of CVI cysts in all cases after delivery. During postnatal follow-up (range 10-48 months), the cyst regressed in one case within 1 month after delivery while the size of the others remained stable. No infant developed psychomotor disorders. CONCLUSIONS: Prenatal sonographic diagnosis of CVI cysts is feasible. Its finding in isolation is consistent with favorable postnatal outcome. PMID- 19565534 TI - Fetal macrosomia is predicted earlier by combination birth-weight estimation methods than by ultrasound alone. PMID- 19565535 TI - Cesarean scar defect: correlation between Cesarean section number, defect size, clinical symptoms and uterine position. AB - OBJECTIVES: To determine the prevalence of clinical symptoms associated with Cesarean scar defects, and to determine the association between the size of these defects, clinical complaints, uterine position, and a history of multiple Cesarean sections. METHODS: In this cross-sectional study, Cesarean scar defects in women with a history of transverse lower-segment Cesarean section were measured by transvaginal ultrasound while being assessed for other gynecological conditions. The relationships between the size of the Cesarean scar defect and the clinical symptoms, uterine position and number of previous Cesarean sections were evaluated. Patients with other uterine pathologies, including endometrial hyperplasia, polyps, malignancy and submucosal myomas, were excluded from the study. RESULTS: During a 3-year period, 4250 women were assessed by transvaginal sonography, of whom 293 (6.9%) were diagnosed with Cesarean scar defects. Eighty six patients were excluded due to other uterine pathologies. Altogether, 207 patients with Cesarean scar defects were included in this study. Women who had undergone multiple Cesarean sections tended to have larger scar defects (in width and depth) than did those who had undergone a single Cesarean section. Women with retroflexed uteri also tended to have wider defects than those with anteflexed uteri. Defect width was significantly greater in women with postmenstrual spotting, dysmenorrhea and chronic pelvic pain. CONCLUSIONS: Multiple Cesarean sections and retroflexed uteri are risk factors for larger Cesarean scar defects. The size of the Cesarean scar defect is associated with clinical symptoms such as postmenstrual spotting, dysmenorrhea and chronic pelvic pain. PMID- 19565536 TI - The role of ultrasound-indicated cerclage in triplets. AB - OBJECTIVES: Preterm delivery is the leading cause of major perinatal morbidity and mortality associated with triplet pregnancies. The objective of this study was to evaluate the efficacy of ultrasound-indicated cervical cerclage in triplet pregnancies that are diagnosed with cervical shortening on biweekly transvaginal sonography (TVS). METHODS: A retrospective review of all triplets who were followed with biweekly TVS for measurement of cervical length was conducted. Cervical shortening was defined as cervical length or =20 years who reported that they had been diagnosed with arthritis or rheumatism by a health professional. Logistic regression analyses were conducted to investigate depression and suicidal ideation. RESULTS: One in 10 Canadians with arthritis had clinically relevant levels of major depression. The age- and sex-adjusted odds ratios (ORs) of major depression (OR 2.24, 95% confidence interval [95% CI] 2.11-2.38) and suicidal ideation (OR 2.01, 95% CI 1.75-2.31) among those with arthritis were approximately twice that of those without arthritis. The adjusted ORs of major depression among those with arthritis were significantly higher among women, the unmarried, younger, and poorer individuals. Individuals in pain, with limitations in activities of daily living, with limitations in instrumental activities of daily living, and with greater numbers of chronic conditions had higher odds of major depression. Less than half of those with major depression had consulted a mental health professional. One in 5 individuals with arthritis and major depression had been suicidal in the past year. CONCLUSION: The majority of individuals with arthritis and major depression were not receiving adequate treatment for major depression. Clients should be screened for major depression and suicidal ideation, particularly if they fall into the identified vulnerable groups. PMID- 19565541 TI - Variables associated with the progression of hip osteoarthritis: a systematic review. AB - OBJECTIVE: As populations age and the prevalence of hip osteoarthritis (OA) increases, health care providers must manage increasing demands for services. Evidence regarding the progression of hip OA can assist health care practitioners in determining expected patient prognosis and planning care. This systematic review of prospective cohort studies examines prognostic variables in patients with hip OA. METHODS: Articles were selected following a comprehensive search of Medline, EMBase, CINAHL, and Allied and Complementary Medicine from database inception to October 2008 and hand searches of the reference lists of retrieved articles. Inclusion criteria involved 1) estimates of the association between prognostic variables and progression of OA, 2) prospective cohort design, 3) patients diagnosed with hip OA based on established criteria, 4) at least 1 year of followup, and 5) access to the full published text. Two independent reviewers assessed the methodologic quality of each study and the association between prognostic variables and OA progression. RESULTS: Eighteen articles met the inclusion criteria; 17 were considered to be of high quality. Strong evidence of progression was associated with age, joint space width at entry, femoral head migration, femoral osteophytes, bony sclerosis, Kellgren/Lawrence hip grade 3, baseline hip pain, and Lequesne index score > or =10. Strong evidence of no association with progression was associated with acetabular osteophytes. Evidence was weak or inconclusive regarding associations between various other radiographic or clinical variables, molecular biomarkers, or use of nonsteroidal inflammatory drugs. CONCLUSION: Overall, few variables were found to be strongly associated with the progression of hip OA, and a variety of other variables were weakly predictive of outcome. PMID- 19565546 TI - Validation of whole-body against conventional magnetic resonance imaging for scoring acute inflammatory lesions in the sacroiliac joints of patients with spondylarthritis. AB - OBJECTIVE: To compare the performance of whole-body magnetic resonance imaging (MRI) versus conventional MRI in assessing acute inflammatory lesions of the sacroiliac (SI) joints in patients with established and active spondylarthritis (SpA) using the Spondyloarthritis Research Consortium of Canada (SPARCC) MRI index. This study is validating whole-body MRI against the current MRI standard for assessing active inflammatory lesions of the SI joints in patients with SpA. METHODS: Thirty-two SpA patients with clinically active disease (Bath Ankylosing Spondylitis Disease Activity Index score >/=4) fulfilling the modified New York criteria were scanned by whole-body and conventional MRI of the SI joints. The MRIs were scored independently in random order by 3 readers blinded to patient identity. Active inflammatory lesions of the SI joints were recorded on a Web based SPARCC index. Pearson's correlation coefficients were used to compare scores for whole-body and conventional MRI for each reader, whereas intraclass correlation coefficients (ICCs) were used to compare interobserver reliability. RESULTS: The Pearson's correlation coefficients between whole-body and conventional MRI per rater were 0.94, 0.87, and 0.93. The mean sum scores for conventional versus whole-body MRI were statistically significantly higher for all 3 readers, although all patients showing inflammatory lesions on conventional MRI also demonstrated them on whole-body MRI. The ICCs(2,1) were 0.69, 0.78, and 0.95 for conventional MRI, and 0.79, 0.85, and 0.96 for whole-body MRI for the 3 possible reader pairs. CONCLUSION: Whole-body and conventional MRI scores show a strong correlation and comparable reliability for the detection of inflammatory lesions of the SI joints. PMID- 19565542 TI - Multicenter, randomized, double-blind, controlled trial of treatment of active rheumatoid arthritis with T-614 compared with methotrexate. AB - OBJECTIVE: To assess the efficacy and safety of T-614 versus methotrexate (MTX) in patients with active rheumatoid arthritis (RA). METHODS: In this multicenter, double-blind trial, 489 patients randomly received either T-614 25 mg/day for the first 4 weeks and 50 mg/day for the subsequent 20 weeks (group 1, n = 163), T-614 50 mg/day for 24 weeks (group 2, n = 163), or MTX 10 mg/week for the first 4 weeks and 15 mg/week for the subsequent 20 weeks (n = 163). Clinical and laboratory parameters were analyzed at baseline and at 4, 10, 17, and 24 weeks. RESULTS: After 24 weeks of treatment, the American College of Rheumatology 20% improvement criteria response rate for patients in T-614 group 2 (63.8%) was not statistically significantly different from that for patients receiving MTX treatment (62.0%), and was superior to that for patients in T-614 group 1 (50.9%). The result of the noninferiority analysis indicated that the efficacy of T-614 (50 mg/day) was not lower than that of MTX by <10%. Rheumatoid factor and IgA, IgG, and IgM demonstrated a statistically significant decrease in all groups. Frequently reported adverse events included hematologic disorder, skin reactions, gastrointestinal symptoms, and transient liver enzyme elevations in the T-614 therapy groups. Side effects in the T-614 groups were generally fewer and milder than in the MTX group, except for skin reactions. There were no prominent cardiovascular adverse events and gastrointestinal ulcers found in the T-614 groups. CONCLUSION: Results indicate that T-614 therapy 50 mg/day is effective and well tolerated, and represents a new option for the treatment of patients with active RA. PMID- 19565547 TI - Feasibility of second-generation ultrasound contrast media in the detection of active sacroiliitis. AB - OBJECTIVE: To determine whether a recently available contrast-enhanced ultrasound (CEUS) technique using second-generation microbubbles allows for the detection of active sacroiliitis, and to measure CEUS enhancement depth at the dorsocaudal part of the sacroiliac (SI) joints in healthy volunteers compared with patients with sacroiliitis. METHODS: Forty-two consecutive patients (84 SI joints) presenting with a clinical diagnosis of sacroiliitis in 50 SI joints and 21 controls (42 SI joints) were investigated by CEUS using a standardized low mechanical index ultrasound protocol. Detected vascularity was used to retrospectively measure the enhancement depth in the dorsocaudal part of the SI joints. RESULTS: CEUS detected enhancement in all clinically active SI joints, showing an enhancement depth into the dorsal SI joint cleft of 18.5 mm (range 16 22.1), which was significantly higher compared with both inactive joints of patients (3.6 mm, range 0-12; P < 0.001) and healthy controls (3.1 mm, range 0 7.8; P < 0.001). All inactive joints were correctly classified based on a lack of deep enhancement in patients with sacroiliitis and controls (42 of 42, 100% sensitivity, 100% specificity; Cohen's kappa = 1). CONCLUSION: CEUS allowed the differentiation of active sacroiliitis from inactive SI joints, and proved to be a feasible method for the detection of vascularity in clinically active sacroiliitis by showing deep contrast enhancement into the SI joints not detectable in inactive joints of patients or controls. If this technique might add information to the earlier detection of sacroiliitis, it should be addressed in further studies. PMID- 19565548 TI - Effect of baseline quadriceps activation on changes in quadriceps strength after exercise therapy in subjects with knee osteoarthritis. AB - OBJECTIVE: To examine whether pretreatment magnitude of quadriceps activation (QA) helps predict changes in quadriceps strength after exercise therapy in subjects with knee osteoarthritis (OA). We hypothesized that subjects with lower magnitudes of QA (greater failure of muscle activation) would have smaller gains in strength compared with those with higher magnitudes of QA following exercise therapy. METHODS: One hundred eleven subjects with knee OA (70 women) participated. Baseline measures included demographic information, quadriceps muscle strength, and QA using a burst-superimposition isometric torque test. Following baseline testing, subjects underwent a 6-week supervised exercise program designed to improve strength, range of motion, balance and agility, and physical function. On completion of the program, quadriceps strength and QA were reassessed. Multiple regression analysis was used to determine whether baseline QA predicted quadriceps strength scores at the 2-month followup. RESULTS: Bivariate correlations demonstrated that baseline QA was significantly associated with quadriceps strength at baseline (rho = 0.30, P < 0.01) and 2-month followup (rho = 0.23, P = 0.01). Greater magnitude of baseline QA correlated with higher strength. While controlling for baseline quadriceps strength and type of exercise therapy, the level of QA did not predict quadriceps strength at the 2-month followup (beta = -0.04, P = 0.18). CONCLUSION: Baseline QA did not predict changes in quadriceps strength following exercise therapy. Measurement of QA using the central activation ratio method does not appear to be helpful in identifying subjects with knee OA who will have difficulty improving quadriceps strength with exercise therapy. PMID- 19565549 TI - Effectiveness of an early cognitive-behavioral treatment in patients with work disability due to musculoskeletal disorders. AB - OBJECTIVE: To evaluate whether an early cognitive-behavioral treatment complementary to a rheumatologic care program, for patients with recent-onset temporary work disability caused by musculoskeletal disorders (MSDs) is effective. METHODS: Patients with an MSD-related temporary work disability episode from 3-8 weeks' duration who were in a rheumatologic care program were randomized into a control group (rheumatologic care program) or an intervention group (rheumatologic care program plus cognitive-behavioral treatment). Enrollment lasted 24 months and followup lasted 6-24 months. Efficacy variables included duration of temporary work disability episodes, total number of work days saved, relative efficacy, and relative rate to return to work. An economic evaluation was also performed. RESULTS: One hundred eighty-one patients were included (66 control and 115 intervention patients), generating 222 episodes of MSD-related temporary work disability. Episodes tended to be shorter in the intervention group than in the control group (mean 98 versus 127 days; P = 0.053), with a relative efficacy of 22.9%. There were no differences in duration of the first episode between groups (mean 105 versus 110 days; P = 0.79), but relapse episodes were significantly shorter in the intervention group (mean 63 days versus 197 days; P = 0.0002). Costs were also lower in the intervention group. To save 1 day of temporary work disability, $13.50 had to be invested in the program. Each dollar invested generated a benefit of $4.08. The program had a net benefit of $172,607. CONCLUSION: Early cognitive-behavioral treatment complementary to a rheumatologic care program is cost-effective, adds >20% efficacy to the rheumatologic care program, and reduces the duration of relapses. PMID- 19565550 TI - Development of a structured interview tool to help patients identify and solve rheumatic condition-related work barriers. AB - OBJECTIVE: To develop a comprehensive and efficient assessment tool for rheumatic condition-related work barriers and explore its use by physical and occupational therapists. METHODS: Literature on arthritis work barriers was examined, followed by the collection of qualitative data on work barriers from patients with rheumatic conditions. A tool called the Work Experience Survey-Rheumatic Conditions (WES-RC), which identifies barriers and facilitates the formation of solutions for barriers, was developed using this data. Ten physical and occupational therapists reviewed the initial version of the tool and provided qualitative data and the relevance of its use by therapists. Using this data, the WES-RC was revised. The therapists then administered the WES-RC to 20 patients. Quantitative data, qualitative data, and results of the administrative experience were collected from therapists after each administration. Relevant data were used to further revise the WES-RC. Qualitative data were coded and themes developed by 2 readers and compared. Means and frequencies were used to describe the quantitative data. RESULTS: The WES-RC addressed patients' work barriers quite well with a mean score of 8.7, on a scale of 1-10 where 10 = barriers completely covered, and administration time was reported as "about right" in 18 (90%) of 20 administrations. Eighty percent of the therapists' administration experiences were positive. Therapists reported barrier identification as easy, while solution formation was difficult in 45% of the administrations and judged insufficient in 35%. CONCLUSION: The WES-RC appears to be feasible for rheumatology patients and for use by physical and occupational therapists. Further study is needed to enhance effective solution formation. PMID- 19565551 TI - Targeted ultrasound of the fifth metatarsophalangeal joint in an early inflammatory arthritis cohort. AB - OBJECTIVE: To determine whether targeted ultrasonographic (US) imaging of the fifth metatarsophalangeal (MTP) joint, compared with radiographs, could aid in the early diagnosis of rheumatoid arthritis (RA) by identifying erosions sooner in early inflammatory arthritis. Radiographic erosion in RA is a late indication of poor prognosis. The earlier detection of erosion may facilitate the timely initiation of disease-modifying antirheumatic drug therapy, particularly in patients with undifferentiated synovitis. METHODS: Patients presenting with synovitis for the first time were invited to participate. Each patient underwent laboratory tests, radiographs of the hands and feet, and US imaging of both fifth MTP joints. RESULTS: Thirty patients (22 women) took part in the study. Seventeen patients (57%) had RA, and 13 (43%) had undifferentiated arthritis (UA). The mean +/- SD time taken to scan both fifth MTP joints was 10.9 +/- 4.4 minutes. Ten patients (33%) had US evidence of synovitis associated with a positive power Doppler (PD) signal (P = 0.04). Seven patients (23%) had radiographic erosions of the fifth MTP joint, and 17 patients (57%) had US evidence of fifth MTP joint erosions (P = 0.01). A positive PD signal at the fifth MTP joint was seen in 9 of 17 patients with RA and 1 of 13 patients with UA (P = 0.02). Patients with a definite diagnosis of RA were more likely to have fifth MTP joint erosions (11 [65%] of 17) compared with UA (6 [46%] of 13). CONCLUSION: Targeted US is a rapid and useful tool in detecting erosive disease in early inflammatory arthritis. It gives a better indication of disease severity and prognosis compared with routinely available laboratory tests, even in the absence of a definite diagnosis. PMID- 19565552 TI - Clinical and immunogenetic prognostic factors for radiographic severity in ankylosing spondylitis. AB - OBJECTIVE: To improve prognostic ability in ankylosing spondylitis (AS), we sought to identify demographic, clinical, and immunogenetic characteristics associated with radiographic severity in a large cohort of patients. METHODS: Patients with AS for > or =20 years were enrolled in a cross-sectional study (n = 398). Pelvic and spinal radiographs were scored using the Bath Ankylosing Spondylitis Radiology Index for the spine (BASRI-s), and radiographic severity was measured as the BASRI-s/duration of AS. Clinical factors and HLA-B, DR, DQ, and DP alleles associated with the highest quartile of the distribution of radiographic severity were identified by first using random forests and then using multivariable logistic regression modeling. Similar procedures were used to identify factors associated with the lowest quartile of radiographic severity. RESULTS: Radiographic severity (being in the top quartile of BASRI-s/duration of AS) was associated with older age at onset of AS (odds ratio [OR] 1.10 per year), male sex (OR 1.90), current smoker (OR 4.72), and the presence of HLA-B*4100 (OR 11.73), DRB1*0804 (OR 12.32), DQA1*0401 (OR 5.24), DQB1*0603 (OR 3.42), and DPB1*0202 (OR 23.36), whereas the presence of DRB1*0801 was strongly negatively associated (OR 0.03). Being in the lowest quartile of BASRI-s/duration of AS was also less likely among those with an older age at onset of AS (OR 0.94 per year), men (OR 0.28), and current smokers (OR 0.29). CONCLUSION: The accuracy of the prognosis of radiographic severity in AS is improved by knowing the age at disease onset, sex, smoking history, and the presence of HLA-B*4100, DRB1*0804, DQA1*0401, DQB1*0603, DRB1*0801, and DPB1*0202 alleles. PMID- 19565553 TI - Anti-U11/U12 RNP antibodies in systemic sclerosis: a new serologic marker associated with pulmonary fibrosis. AB - OBJECTIVE: To characterize a new serum autoantibody in patients with systemic sclerosis (SSc) directed against U11/U12 RNP and to identify the clinical features associated with this autoantibody. METHODS: We identified autoantibodies directed against the U11/U12 RNP complex in sera of patients with SSc and confirmed antibody specificity by immunoprecipitation, reverse transcriptase polymerase chain reaction, and Southern blotting. We determined the prevalence of these antibodies in SSc and their specificity for SSc. We compared anti-U11/U12 RNP autoantibody-positive and negative SSc patients on demographic, disease classification, clinical variables, and survival. RESULTS: We identified 33 patients with anti-U11/U12 RNP antibodies. In 2 consecutive series of SSc patients first seen at 10-year intervals (1994-1995 and 2004-2005), the prevalence of anti-U11/U12 RNP antibody-positive patients was 15 of 462 (3.2%). Seventeen (52%) of these 33 patients had limited cutaneous involvement. All patients had Raynaud's phenomenon and 82% had gastrointestinal (GI) involvement. None had "intrinsic" pulmonary arterial hypertension. The most significant clinical difference between anti-U11/U12 antibody-positive and negative cohorts was the prevalence of lung fibrosis, which occurred in 79% of the anti-U11/U12 RNP antibody-positive patients versus 37% of the anti-U11/U12 RNP antibody negative patients (P < 0.0001). GI involvement was also significantly increased in the anti-U11/U12 RNP antibody-positive group. Patients with anti-U11/U12 RNP antibodies and pulmonary fibrosis had a 2.25-fold greater risk of death than anti U11/U12 RNP negative patients with pulmonary fibrosis. CONCLUSION: Anti-U11/U12 RNP antibodies are present in the sera of approximately 3% of patients with SSc and are a marker for lung fibrosis, which is often severe. PMID- 19565554 TI - Mail-delivered arthritis self-management tool kit: a randomized trial and longitudinal followup. AB - OBJECTIVE: To determine the effectiveness of an intervention Tool Kit of arthritis self-management materials to be sent once through the mail, and to describe the populations reached. METHODS: Spanish speakers (n = 335), non Hispanic English-speaking African Americans (n = 156), and other non-Hispanic English speakers (n = 404) were recruited separately and randomized within each of the 3 ethnic/racial categories to immediately receive the intervention Tool Kit (n = 458) or to a 4-month wait-list control status (n = 463). At the end of 4 months, controls were sent the Tool Kit. All subjects were followed in a longitudinal study for 9 months. Self-administered measures included health status, health behavior, arthritis self-efficacy, medical care utilization, and demographic variables. Using analyses of covariance and t-tests, analyses were conducted for all participants and for Spanish- and English-language groups. RESULTS: At 4 months, comparing all intervention subjects with randomized wait list controls, there were significant (P < 0.01) benefits in all outcomes except medical care utilization and self-rated health. The results were maintained at 9 months compared with baseline. On average, the Tool Kit reached persons ages 50 56 years with 12-15 years of schooling. There were few differences between English- and Spanish-language participants in either the effectiveness or reach variables. CONCLUSION: A mailed Arthritis Self-Management Tool Kit proved effective in improving health status, health behavior, and self-efficacy variables for up to 9 months. It also reached younger persons in both English- and Spanish-language groups and Spanish speakers with higher education levels than previous studies of the small-group Arthritis Self-Management Program. PMID- 19565555 TI - Being careful: a grounded theory of emergent chronic knee problems. AB - OBJECTIVE: To gain insight into the prediagnostic stages of knee osteoarthritis (OA) and identify the process whereby people recognize and address emergent chronic knee problems. METHODS: Twenty-six people (15 women, mean age 53.2 +/- 7.4 years) participated in a grounded theory study. Ten participants had a recent diagnosis of knee OA, and 16 had no diagnosis. The undiagnosed participants self reported their symptoms, which had lasted at least 6 months and were consistent with knee OA. During semistructured, one-on-one interviews, participants reflected on the development and impact of their chronic knee problems. A constant comparative approach was used for analysis. RESULTS: Participants described uncertainty in understanding the meaning of intermittent knee symptoms for several years before becoming aware of the emergence of chronic knee problems. Once aware, participants engaged in a circular process of interpreting the meaning of knee symptoms and being careful. Being careful referred to the cycle of perceptions, intentions, and behaviors aimed at avoiding knee damage during physical activity. This cycle continued until participants experienced a disruption that challenged their participation in meaningful activities, at which time they decided to access health care. CONCLUSION: As a new construct, being careful unifies the complex set of experiences and behaviors that describe how participants protected their knee during physical activity. Participants interpret the experiences associated with emerging knee problems through interactions with others. These interactions enhance the participants' self management, despite not having the benefits associated with diagnosis, such as justification for symptoms and formal assistance. PMID- 19565557 TI - Does the use of ordered values of subregional change in cartilage thickness improve the detection of disease progression in longitudinal studies of osteoarthritis? AB - OBJECTIVE: To propose a novel strategy for more efficiently measuring changes in cartilage thickness in osteoarthritis (OA) using magnetic resonance imaging, and to hypothesize that determining the magnitude of thickness change independent of the anatomic location provides improved discrimination between healthy subjects and OA participants longitudinally. METHODS: A total of 148 women were imaged; 90 were Kellgren/Lawrence (K/L) grade 0, 30 were K/L grade 2, and 28 were K/L grade 3. Magnetic resonance images (3T) were acquired at baseline and at 24 months. Changes in femorotibial cartilage thickness were determined in 5 tibial and 3 femoral medial and lateral subregions, respectively (conventional approach). The new strategy provided ordered values of subregional change in each compartment, ranked according to the direction and magnitude of change. RESULTS: Using the new ordered values approach, the minimal P value for the differences in 2-year change in medial cartilage thickness of K/L grade 3 and K/L grade 0 participants was 0.001 (Wilcoxon test), with 4 ordered medial subregions differing significantly between both groups. With the conventional approach, only 1 medial subregion differed significantly between K/L grade 3 and K/L grade 0 (P = 0.037). Cartilage thickening was significantly greater in K/L grade 2 versus K/L grade 0 participants in 1 medial subregion using the conventional approach (P = 0.016), and in 2 medial subregions (minimal P = 0.007) using the ordered values approach. CONCLUSION: The novel ordered values approach is more sensitive in detecting cartilage thinning in K/L grade 3 and cartilage thickening in K/L grade 2 versus K/L grade 0 participants. The new method may be particularly useful in the context of other comparisons, e.g., a group treated with a disease-modifying OA drug versus one treated with a placebo. PMID- 19565556 TI - Hyperuricemia and risk of stroke: a systematic review and meta-analysis. AB - OBJECTIVE: To assess the association between hyperuricemia and risk of stroke incidence and mortality because hyperuricemia is hypothesized to be a risk factor for stroke and other cardiovascular disease, but, to date, results from observational studies are conflicting. METHODS: A systematic review and meta analysis were conducted. Studies were identified by searching major electronic databases using the Medical Subject Headings and keywords without restriction in languages. Prospective cohort studies were included only if they contained data on stroke incidences or mortalities related to serum uric acid levels in adults. Pooled risk ratios (RRs) for the association of stroke incidence and mortality with serum uric acid levels were calculated. RESULTS: A total of 16 studies including 238,449 adults were eligible and abstracted. Hyperuricemia was associated with a significantly higher risk of both stroke incidence (6 studies; RR 1.41, 95% confidence interval [95% CI] 1.05, 1.76) and mortality (6 studies; RR 1.36, 95% CI 1.03, 1.69) in our meta-analyses of unadjusted study estimates. Subgroup analyses of studies adjusting for known risk factors such as age, hypertension, diabetes mellitus, and cholesterol still showed that hyperuricemia was significantly associated with both stroke incidence (4 studies; RR 1.47, 95% CI 1.19, 1.76) and mortality (6 studies; RR 1.26, 95% CI 1.12, 1.39). The pooled estimate of multivariate RRs did not differ significantly by sex. CONCLUSION: Hyperuricemia may modestly increase the risks of both stroke incidence and mortality. Future research is needed to determine whether lowering uric acid level has any beneficial effects on stroke. PMID- 19565558 TI - Sensitivity and specificity of spinal inflammatory lesions assessed by whole-body magnetic resonance imaging in patients with ankylosing spondylitis or recent onset inflammatory back pain. AB - OBJECTIVE: To determine the diagnostic utility of different spinal inflammatory lesions assessed by whole-body magnetic resonance imaging (MRI) in patients with ankylosing spondylitis (AS) or with recent-onset inflammatory back pain (IBP) compared with healthy controls. METHODS: We scanned 35 consecutive patients with AS fulfilling the modified New York criteria, 25 patients with IBP of <24 months' duration (both groups were age < or =45 years and had a Bath Ankylosing Spondylitis Disease Activity Index score > or =4), and 35 healthy age- and sex matched volunteers using whole-body MRI STIR sequences of the spine. MRIs were independently assessed in random order by 3 readers blinded to patient identity. Inflammatory spinal lesions were recorded consistent with definitions proposed by the Canada/Denmark International MRI Working Group: vertebral corner inflammatory lesions (CIL) and noncorner inflammatory lesions in central sagittal slices and lateral inflammatory lesions (LIL) in lateral slices. Concordantly scored lesions for the 3 possible reader pairs were used in the analysis of sensitivity, specificity, likelihood ratios (LRs), and areas under the curve for the entire spine and by spinal segment. RESULTS: Diagnostic utility was optimal when > or =2 CIL were recorded (for patients with AS, values for sensitivity, specificity, and positive LR were 69%, 94%, and 12, respectively, and for patients with IBP were 32%, 96%, and 8, respectively). LIL had high specificity (97%) but low sensitivity (31%). Nine controls had > or =1 CIL, but only 2 controls had >2 CIL. CONCLUSION: Diagnostic utility of STIR MRI for AS is optimal when > or =2 CIL are present. A single CIL can be found in up to 26% of healthy individuals. PMID- 19565559 TI - Efficacy of intraarticular infliximab in patients with chronic or recurrent gonarthritis: a clinical randomized trial. AB - OBJECTIVE: To evaluate the efficacy and safety of intraarticular infliximab compared with intraarticular methylprednisolone in patients with gonarthritis. METHODS: In 23 patients with recurrent gonarthritis despite previous intraarticular corticosteroid therapy, a total of 41 intraarticular injections (20 infliximab and 21 methylprednisolone) were performed in 28 knees. Initial therapy was randomly assigned, and crossover therapy was eligible within 3 months. The clinical effect was assessed during 6 months of followup. The primary outcome was event-free survival, defined as the time after treatment until local retreatment was performed and/or nonimprovement of the knee joint score. Adverse effects were recorded during followup. RESULTS: All patients treated with intraarticular infliximab had an insufficient response. In contrast, 8 of the 21 intraarticular methylprednisolone injections were effective (P = 0.004). Between groups, no differences in the patients' age, disease duration, number of disease modifying antirheumatic drugs, or previous intraarticular methylprednisolone were observed. Reported adverse effects were not related to therapy. CONCLUSION: Treatment with intraarticular infliximab injection was not effective in patients with a chronically inflamed knee joint. Intraarticular injection with methylprednisolone was superior despite previous intraarticular corticosteroid therapy. Further investigation is needed to provide these patients with a better alternative. PMID- 19565561 TI - Hypoxia-induced expression of bradykinin type-2 receptors in endothelial cells triggers NO production, cell migration, and angiogenesis. AB - Bradykinin receptors are differentially expressed in the coronary vascular endothelium of rat and human hearts during the pathogenesis of heart failure, but the mechanisms responsible for this regulation have remained vague. Here we show by quantitative real-time PCR, Western blot analysis, and immunohistochemistry, that hypoxia triggers the expression of bradykinin type-2 receptors (BK-2Rs) in cultured human coronary artery endothelial cells (HCAECs), in isolated rat cardiac microvascular endothelial cells (RCMECs), and in rat hearts subjected to ligation of the left anterior descending coronary artery. Mild hypoxia (5% O(2)) induced a fourfold temporal increase in BK-2R mRNA expression in HCAECs, which was also observed at the protein level, whereas severe hypoxia (1% O(2)) slightly inhibited the mRNA expression of BK-2Rs. In addition, HOE-140, a BK-2R antagonist, inhibited mRNA and protein expression of BK-2Rs. The BK-2Rs induced by mild hypoxia were biologically active, that is, capable of inducing intracellular production of nitric oxide (NO) upon activation of HCAECs with bradykinin (BK), a response attenuated by HOE-140. In rat hearts recovering from myocardial infarction, BK-2Rs were upregulated in the endothelium of vessels forming at the border zone between fibrotic scar tissue and healthy myocardium. Furthermore, in an in vitro wound-healing assay, RCMEC migration was increased under mild hypoxic culture conditions in the presence of BK and was attenuated with HOE-140. Our present results show that mild hypoxia triggers a temporal expression of functional BK-2Rs in human and rat endothelial cells and support a role for BK-2Rs in hypoxia-induced angiogenesis. PMID- 19565560 TI - Comparison of two methods of conducting the Fit and Strong! program. AB - OBJECTIVE: Fit and Strong! is an award winning, evidence-based, multiple component physical activity/behavior change intervention. It is a group- and facility-based program that meets for 90 minutes 3 times per week for 8 weeks (24 sessions total). We originally tested Fit and Strong! using physical therapists (PTs) as instructors but have transitioned to using nationally certified exercise instructors (CEIs) as part of an effort to translate Fit and Strong! into community-based settings, and have tested the impact of this shift in instruction type on participant outcomes. METHODS: We used a 2-group design. The first 161 participants to sequentially enroll received instruction from PTs. The next 190 sequential enrollees received instruction from CEIs. All participants were assessed at baseline, at the conclusion of the 8-week Fit and Strong! program, and at the 6-month followup. RESULTS: We saw no significant differences by group on outcomes at 8 weeks or 6 months. Participants in both groups improved significantly with respect to lower-extremity strength, aerobic capacity, pain, stiffness, and physical function. Significant differences favoring the PT-led classes were seen on 2 of 5 mediators, self-efficacy for exercise and barriers adherence efficacy. Participant evaluations rated both types of instruction equally highly, attendance was identical, and no untoward health events were observed or reported under either instruction mode. CONCLUSION: Outcomes under the 2 types of instruction are remarkably stable. These findings justify the use of CEIs in the future to extend the reach of the Fit and Strong! program. PMID- 19565562 TI - Nonparametric equivalence testing with respect to the median difference. AB - The problem of comparing two independent groups of univariate data in the sense of testing for equivalence is considered for a fully nonparametric setting. The distribution of the data within each group may be a mixture of both a continuous and a discrete component, and no assumptions are made regarding the way in which the distributions of the two groups of data may differ from each other - in particular, the assumption of a shift model is avoided. The proposed equivalence testing procedure for this scenario refers to the median of the independent difference distribution, i.e. to the median of the differences between independent observations from the test group and the reference group, respectively. The procedure provides an asymptotic equivalence test, which is symmetric with respect to the roles of 'test' and 'reference'. It can be described either as a two-one-sided-tests (TOST) approach, or equivalently as a confidence interval inclusion rule. A one-sided variant of the approach can be applied analogously to non-inferiority testing problems. The procedure may be generalised to equivalence testing with respect to quantiles other than the median, and is closely related to tolerance interval type inference. PMID- 19565563 TI - miR-29 suppression of osteonectin in osteoblasts: regulation during differentiation and by canonical Wnt signaling. AB - The matricellular protein osteonectin, secreted protein acidic and rich in cysteine (SPARC, BM-40), is the most abundant non-collagenous matrix protein in bone. Matricellular proteins play a fundamental role in the skeleton as regulators of bone remodeling. In the skeleton, osteonectin is essential for the maintenance of bone mass and for balancing bone formation and resorption in response to parathyroid hormone (PTH). It promotes osteoblast differentiation and cell survival. Mechanisms regulating the expression of osteonectin in the skeleton and in other tissues remain poorly understood. We found that the proximal region of the mouse osteonectin 3' untranslated region (UTR) contains a well-conserved, dominant regulatory motif that interacts with microRNAs (miRs) 29a and -29c. Transfection of osteoblastic cells with miR-29a inhibitors increased osteonectin protein levels, whereas transfection of miR-29a precursor RNA decreased osteonectin. miR-29a and -29c were increased during osteoblastic differentiation in vitro. The up-regulation of these miRNAs correlated with decreased osteonectin protein during the matrix maturation and mineralization phases of late differentiation. In contrast, osteonectin transcript levels remained relatively constant during this process, implying repression of translation. Treatment of osteoblasts with LiCl induced miR-29a and -29c expression and decreased osteonectin synthesis. When cells were treated with Dickkopf-1 (Dkk-1), miR-29a and -29c expression was repressed. These data suggest that canonical Wnt signaling, which is increased during osteoblastic differentiation, induces expression of miR-29. Osteonectin and miR-29 are co expressed in extra-skeletal tissues, and the post-transcriptional mechanisms regulating osteonectin in osteoblasts are likely to be active in other cell systems. PMID- 19565564 TI - Eupatilin inhibits T-cell activation by modulation of intracellular calcium flux and NF-kappaB and NF-AT activity. AB - Eupatilin, one of the pharmacologically active ingredients of Artemisia princeps, exhibits a potent anti-ulcer activity, but its effects on T-cell immunity have not been investigated. Here, we show that eupatilin has a profound inhibitory effect on IL-2 production in Jurkat T cells as well as in human peripheral blood leukocytes. Eupatilin neither influenced clustering of CD3 and LFA-1 to the immunological synapse nor inhibited conjugate formation between T cells and B cells in the presence or absence of superantigen (SEE). Eupatilin also failed to inhibit T-cell receptor (TCR) internalization, thereby, suggesting that eupatilin does not interfere with TCR-mediated signals on the membrane proximal region. In unstimulated T cells, eupatilin significantly induced apoptotic cell death, as evidenced by an increased population of annexin V(+)/PI(+) cells and cleavage of caspase-3 and PARP. To our surprise, however, once cells were activated, eupatilin had little effect on apoptosis, and instead slightly protected cells from activation-induced cell death, suggesting that apoptosis also is not a mechanism for eupatilin-induced T-cell suppression. On the contrary, eupatilin dramatically inhibited I-kappaBalpha degradation and NF-AT dephosphorylation and, consequently, inhibited NF-kappaB and NF-AT promoter activities in PMA/A23187 stimulated T cells. Interestingly, intracellular calcium flux was significantly perturbed in cells pre-treated with eupatilin, suggesting that calcium-dependent cascades might be targets for eupatilin action. Collectively, our results provide evidence for dual regulatory functions of eupatilin: (1) a pro-apoptotic effect on resting T cells and (2) an immunosuppressive effect on activated T cells, presumably through modulation of Ca(2+) flux. PMID- 19565565 TI - Deregulated NFATc1 activity transforms murine fibroblasts via an autocrine growth factor-mediated Stat3-dependent pathway. AB - The nuclear factor of activated T cells (NFAT) family of transcription factors has recently been implicated with a role in tumorigenesis. Forced expression of a constitutively active NFATc1 mutant (caNFATc1) has been shown to transform immortalized murine fibroblasts in vitro, while constitutive activation of the NFAT-signaling pathway has been found in a number of human cancers, where it has been shown to contribute towards various aspects of the tumor phenotype. Here we have investigated the molecular mechanisms underlying the oncogenic potential of deregulated NFAT activity. We now show that ectopic expression of caNFATc1 in murine 3T3-L1 fibroblasts induces the secretion of an autocrine factor(s) that is sufficient to promote the transformed phenotype. We further demonstrate that this NFATc1-induced autocrine factor(s) specifically induces the tyrosine phosphorylation of the Stat3 transcription factor via a JAK kinase-dependent pathway. Interestingly, this effect of sustained NFAT signaling on the autocrine growth factor-mediated activation of Stat3 is not restricted to murine fibroblasts, but is also observed in the PANC-1 and MCF10A human cell lines. Most importantly, we find that the shRNA-mediated depletion of endogenous Stat3 significantly attenuates the ability of caNFATc1 to transform 3T3-L1 fibroblasts. Taken together, our results afford significant new insights into the molecular mechanisms underlying the oncogenic potential of deregulated NFATc1 activity by demonstrating that constitutive NFATc1 activity transforms cells via an autocrine factor-mediated pathway that is critically dependent upon the activity of the Stat3 transcription factor. PMID- 19565566 TI - Zebrafish blood stem cells. AB - Within the past two decades, the zebrafish (Danio rerio) has become an excellent model to study the development of hematopoietic stem cells (HSCs). All vertebrates including zebrafish have primitive and definitive waves of hematopoiesis, but self-renewing pluripotent HSCs are only produced by the definitive wave. The primitive wave occurs in two intraembryonic locations called the intermediate cell mass (ICM) and the anterior lateral mesoderm (ALM). Primitive erythropoiesis is in the ICM, whereas myelopoiesis initiates in the ALM. After circulation starts at 24 h post-fertilization, hematopoiesis shifts to the posterior blood island (PBI) for a brief period. The definitive wave starts in the aorta-gonad-mesonephros (AGM). There are three different HSC migration and colonization events that begin 2 days post-fertilization: AGM progenitor cells migrate to (1) the caudal hematopoietic tissue (CHT), which is an intermediate site of blood development; (2) the thymus, which is a site of lymphocyte maturation; and (3) the developing kidney marrow, which is the larval and adult location for production of all hematopoietic cell types, and is comparable to the bone marrow of mammals. Many of the transcription factors and signaling pathways that regulate the formation of HSCs in a zebrafish are conserved with mammals. Large-scale forward and reverse genetic screens have identified zebrafish blood and HSC mutants that represent models for known human diseases. Along with the technological advancements in the field of zebrafish research, future HSC studies in zebrafish will help us illuminate the genetic network controlling the development and function of stem cells in all vertebrates. PMID- 19565567 TI - DNA methylation-mediated epigenetic control. AB - During differentiation and development cells undergo dramatic morphological and functional changes without any change in the DNA sequence. The underlying changes of gene expression patterns are established and maintained by epigenetic processes. Early mechanistic insights came from the observation that gene activity and repression states correlate with the DNA methylation level of their promoter region. DNA methylation is a postreplicative modification that occurs exclusively at the C5 position of cytosine residues (5mC) and predominantly in the context of CpG dinucleotides in vertebrate cells. Here, three major DNA methyltransferases (Dnmt1, 3a, and 3b) establish specific DNA methylation patterns during differentiation and maintain them over many cell division cycles. CpG methylation is recognized by at least three protein families that in turn recruit histone modifying and chromatin remodeling enzymes and thus translate DNA methylation into repressive chromatin structures. By now a multitude of histone modifications have been linked in various ways with DNA methylation. We will discuss some of the basic connections and the emerging complexity of these regulatory networks. PMID- 19565568 TI - DNA methylation and histone modification regulate silencing of OPG during tumor progression. AB - The identification of molecules that are down-regulated in malignant phenotype is important for understanding tumor biology and their role in tumor suppression. We compared the expression profile of four normal nasal mucosal (NNM) epithelia and a series of nasopharyngeal cancinoma (NPC) cell lines using cDNA microarray and confirmed the actual expression of the selected genes, and found osteoprotegerin (OPG) to be ubiquitously deficient in NPC cells. We also found OPG to be down regulated in various cancer cell lines, including oral, cervical, ovarian, lung, breast, pancreas, colon, renal, prostate cancer, and hepatoma. Administration of recombinant OPG (rOPG) brought about a reduction in cancer cell growth through apoptotic mechanism. We generated eleven monoclonal antibodies (MAbs) against OPG to study OPG's expression and biological functions in cancer cells. OPG was detected in the tumor stromal regions, but not in the cancer cell per se in surgical specimens of liver cancer. Quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) revealed that OPG was down-regulated in NPC tissues compared with normal nasal polyp (NNP) tissues. In addition, we showed OPG silencing to be associated with promoter methylation as well as histone modifications. In OPG-silenced cancer cell lines, the OPG gene promoter CpG dinucleotides were highly methylated. Compared to normal cells, silenced OPG gene in cancer cells were found to have reduced histone 3 lysine 4 tri-methylation (H3K4me3) and increased histone 3 lysine 27 tri-methylation (H3K27me3). Taken together, these results suggest that OPG silencing in carcinoma cancer cells occurs through epigenetic repression. PMID- 19565569 TI - Comparison of insulin detemir and insulin glargine using a basal-bolus regimen in a randomized, controlled clinical study in patients with type 2 diabetes. AB - BACKGROUND: This treat-to-target study compared the efficacy and safety of insulin detemir (IDet) and insulin glargine (IGla) in a basal-bolus (insulin aspart) regimen in type 2 diabetes. METHODS: 385 patients were randomized 2 : 1 (IDet : IGla). Non-inferiority of IDet to IGla was determined by HbA(1c) 95% CI upper limit <0.4. RESULTS: IDet and IGla showed similar efficacy in HbA(1c) reduction at 26 weeks, as the non-inferiority criterion was met at 26 weeks (LS mean [Det-Gla]: 0.207; 95% CI: 0.0149,0.3995). It appeared that IGla in some cases did better than IDet in terms of HbA(1c), but the difference (0.207%) was not clinically meaningful. Based on the CONSORT guideline, non-inferiority analysis using the LOCF approach was inconclusive regarding possible inferiority of delta 0.4 (LS mean of [Det-Gla]: 0.307; 95% CI: 0.1023, 0.5109). HbA(1c) decreased significantly from baseline in IDet (-1.1% [26 weeks], -0.9% [LOCF], p < 0.001) and in IGla (-1.3% [26 weeks, LOCF], p < 0.001). Final HbA(1c) were 7.1% (26 weeks) and 7.3% (LOCF) in IDet, and 6.9% (26 weeks) and 7.0% (LOCF) in IGla. Final FPG were 130 mg/dL (26 weeks) and 135 mg/dL (LOCF) in IDet, and 134 mg/dL (26 weeks) and 137 mg/dL (LOCF) in IGla. There was significantly less weight gain in IDet-treated patients (1.2 +/- 3.96 kg versus 2.7 +/- 3.94 kg, p = 0.001). Hypoglycemia risk was comparable between groups. The majority of IDet-treated patients (87.4%) remained on a once-daily basal insulin regimen throughout the study. CONCLUSIONS: IDet and IGla were both effective and safe treatments for glycemic control in a basal-bolus regimen for type 2 diabetes. Clinically significant reductions in HbA(1c) were achieved in both groups, but with significantly less weight gain in the IDet group at comparable basal insulin dosage. PMID- 19565570 TI - The cell adhesion nectin-like molecules (Necl) 1 and 4 suppress the growth and tumorigenic ability of colon cancer cells. AB - A key step in human colon cancer development includes the hyperactivation of Wnt/beta-catenin signaling and the induction of beta-catenin-TCF target genes that participate in colon cancer progression. Recent studies identified members of the immunoglobulin-like cell adhesion molecules (IgCAM) of the L1CAM family (L1 and Nr-CAM) as targets of beta-catenin-TCF signaling in colon cancer cells. L1 was detected at the invasive front of colon cancer tissue and confers metastasis when overexpressed in cells. In contrast to L1, we did not detect in colon cancer cells significant levels of another IgCAM family of molecules, the nectin-like (Necl) receptors Necl1 and Necl4, while Necl4 was previously found in the normal small intestine and colon tissues. We studied the properties of colon cancer cells in which Necl4 and Necl1 were expressed either alone, or in combination, and found that such cells display a wide range of properties associated with tumor suppression. Expression of both Necl1 and Necl4 was the most efficient in suppressing the tumorigenicity of colon cancer cells. This was associated with enhanced rates of apoptosis and change in several apoptosis related markers. In contrast to its capacity to suppress tumorigenesis, Necl4 was unable to affect the highly malignant and metastatic capacities of colon cancer cells in which L1 was overexpressed. Our results suggest that various IgCAM receptor families play different roles in affecting the tumorigenic function of the same cells, and that Necl1 and Necl4 can fulfill a tumor suppressive role. PMID- 19565572 TI - Setting of care modifies risk of nursing home placement for older adults with dementia. AB - OBJECTIVE: The purpose of this study was to examine risk of nursing home (NH) placement among older adults receiving publicly funded home and community-based services (HCBS) or assisted living (AL) and to explore whether these settings of care modify the relationship between dementia and risk of NH placement. METHODS: The sample consisted of dually eligible Medicare and Medicaid beneficiaries age 65 and older who received HCBS (n = 1630) or resided in AL (n = 836) in Florida between July 1999 and June 2000. Cox proportional hazards regression was used to estimate risk of NH placement over a 5-year study period and to test the interaction of setting of care by dementia status. RESULTS: In all, 15% of HCBS participants were placed in a NH compared to 26% of AL participants. As indicated by a significant interaction term in the regression model, setting of care modified the relationship between dementia and NH placement (HR = 0.45, CI = 0.31 0.66). In post hoc analyses stratified by setting of care, dementia was associated with a 50% increased risk of NH placement from HCBS (HR = 1.50, CI = 1.12-2.02) but was not associated with placement from AL (HR = 0.86, CI = 0.63 1.16). CONCLUSION: The findings suggest that differences in care provided in HCBS and AL may influence subsequent NH placement for older adults with dementia. PMID- 19565571 TI - Threonine 393 of beta-catenin regulates interaction with Axin. AB - CK2 is a regulatory kinase implicated in embryonic development and in cancer. Among the CK2 substrates is beta-catenin, a protein with dual function in Wnt signaling and cell adhesion. Previously, we reported that CK2 activity is required for beta-catenin stability and we identified threonine (T) 393 as a major CK2 phosphorylation site in beta-catenin. However, it is not known whether phosphorylation at T393 increases beta-catenin stability and if so, what is the mechanism. In this study we investigate the molecular mechanism of beta-catenin stabilization through phosphorylation at T393. We found that pseudophosphorylation of beta-catenin at T393 resulted in a stable activated form of beta-catenin with decreased affinity for Axin in vitro. This phosphomimetic mutant also displayed decreased regulation by Axin in vivo in a bioassay in Xenopus laevis embryos. In contrast, the binding of T393 pseudophosphorylated beta-catenin to E-cadherin was unaffected. Further analysis showed that pseudophosphorylation at T393 did not prevent beta-catenin phosphorylation by GSK3beta. Interestingly, we found that in the presence of pseudophophorylated beta-catenin and another activated form of beta-catenin, the recruitment of GSK3beta to Axin is enhanced. These findings indicate that phosphorylation of T393 by CK2 may affect the stability of beta-catenin through decreased binding to Axin. In addition, the increased recruitment of GSK3beta to the destruction complex in the presence of activated beta-catenin mutants could be a feedback mechanism to suppress overactive Wnt signaling. PMID- 19565576 TI - SERS microscopy: nanoparticle probes and biomedical applications. AB - Surface-enhanced Raman scattering (SERS) microscopy is a novel method of vibrational microspectroscopic imaging for the selective detection of biomolecules in targeted research. This technique combines the advantages of biofunctionalized metal nanoparticles and Raman microspectroscopy for visualizing and quantifying the distribution of target molecules such as proteins in cells and tissues. Advantages of SERS over existing labeling approaches include the tremendous multiplexing capacity, quantification using the characteristic SERS signatures and high photostability. This review summarizes current designs of nanoparticle-based SERS probes and highlights first biomedical applications of SERS microscopy for protein localization ex and in vivo. PMID- 19565573 TI - Diagnosis and staging of mild cognitive impairment, using a modification of the clinical dementia rating scale: the mCDR. AB - OBJECTIVE: To examine the reliability and validity of the mCDR, a modified version of the clinical dementia rating (CDR) scale. METHODS: The mCDR is an informant-based, technician-administered, structured interview with multiple choice responses, which does not include objective cognitive testing. Subjects (n = 556) with no cognitive impairment (NCI), amnestic mild cognitive impairment (aMCI), and dementia were assessed with mCDR, CDR, and neuropsychological evaluation, while medial temporal atrophy (MTA) was measured on MRI scans. The mCDR and CDR were compared with respect to inter-rater reliability, validity, and ability to predict progression in cognitive diagnosis at 12 month follow-up. RESULTS: The mCDR can be administered in less than one third of the time required to administer the CDR (30 min). Inter-rater reliability (Cohen's weighted kappa) was 0.86 for the mCDR and 0.56 for the CDR. Ability to distinguish between NCI, aMCI, and Dementia subjects, and correlations to memory and non-memory measures were marginally better for the CDR, in comparison to the mCDR. Correlations of mCDR and CDR scores to MTA scores did not differ. Baseline mCDR scores predicted transition from NCI to aMCI, whereas baseline CDR scores predicted transition from aMCI to Dementia. CONCLUSIONS: The mCDR, as compared to the CDR, is briefer and more reliable, and is a valid measure of functional ability among subjects with normal cognition, mild cognitive impairment, and mild dementia. The mCDR should be particularly useful as a reliable and economical instrument for assessing change in functional abilities, especially in multi-center clinical trials and population studies of MCI and mild dementia. PMID- 19565577 TI - Site-dependent excited-state dynamics of a fluorescent probe bound to avidin and streptavidin. AB - The excited-state dynamics of biotin-spacer-Lucifer-Yellow (LY) constructs bound to avidin (Avi) and streptavidin (Sav) was investigated using femtosecond spectroscopy. Two different locations in the proteins, identified by molecular dynamics simulations of Sav, namely the entrance of the binding pocket and the protein surface, were probed by varying the length of the spacer. A reduction of the excited-state lifetime, stronger in Sav than in Avi, was observed with the long spacer construct. Transient absorption measurements show that this effect originates from an electron transfer quenching of LY, most probably by a nearby tryptophan residue. The local environment of the LY chromophore could be probed by measuring the time-dependent polarisation anisotropy and Stokes shift of the fluorescence. Substantial differences in both dynamics were observed. The fluorescence anisotropy decays analysed by using the wobbling-in-a-cone model reveal a much more constrained environment of the chromophore with the short spacer. Moreover, the dynamic Stokes shift is multiphasic in all cases, with a approximately 1 ps component that can be ascribed to diffusive motion of bulk like water molecules, and with slower components with time constants varying not only with the spacer, but with the protein as well. These slow components, which depend strongly on the local environment of the probe, are ascribed to the motion of the hydration layer coupled to the conformational dynamics of the protein. PMID- 19565578 TI - Characterization of the biomechanical properties of T4 pili expressed by Streptococcus pneumoniae--a comparison between helix-like and open coil-like pili. AB - Bacterial adhesion organelles, known as fimbria or pili, are expressed by gram positive as well as gram-negative bacteria families. These appendages play a key role in the first steps of the invasion and infection processes, and they therefore provide bacteria with pathogenic abilities. To improve the knowledge of pili-mediated bacterial adhesion to host cells and how these pili behave under the presence of an external force, we first characterize, using force measuring optical tweezers, open coil-like T4 pili expressed by gram-positive Streptococcus pneumoniae with respect to their biomechanical properties. It is shown that their elongation behavior can be well described by the worm-like chain model and that they possess a large degree of flexibility. Their properties are then compared with those of helix-like pili expressed by gram-negative uropathogenic Escherichia coli (UPEC), which have different pili architecture. The differences suggest that these two types of pili have distinctly dissimilar mechanisms to adhere and sustain external forces. Helix-like pili expressed by UPEC bacteria adhere to host cells by single adhesins located at the distal end of the pili while their helix-like structures act as shock absorbers to dampen the irregularly shear forces induced by urine flow and to increase the cooperativity of the pili ensemble, whereas open coil-like pili expressed by S. pneumoniae adhere to cells by a multitude of adhesins distributed along the pili. It is hypothesized that these two types of pili represent different strategies of adhering to host cells in the presence of external forces. When exposed to significant forces, bacteria expressing helix-like pili remain attached by distributing the external force among a multitude of pili, whereas bacteria expressing open coil-like pili sustain large forces primarily by their multitude of binding adhesins which presumably detach sequentially. PMID- 19565579 TI - A novel heavy-atom label for side-specific peptide iodination: synthesis, membrane incorporation and X-ray reflectivity. AB - Structural parameters, such as conformation, orientation and penetration depth of membrane-bound peptides and proteins that may function as channels, pores or biocatalysts, are of persistent interest and have to be probed in the native fluid state of a membrane. X-ray scattering in combination with heavy-atom labeling is a powerful and highly appropriate method to reveal the position of a certain amino acid residue within a lipid bilayer with respect to the membrane normal axis up to a resolution of several Angstrom. Herein, we report the synthesis of a new iodine-labeled amino acid building block. This building block is intended for peptide incorporation to provide high intensities for electron density difference analysis of X-ray reflectivity data and improve the labeling potential for the lipid bilayer head-group and water region. The novel building block as well as the commercially available non-iodinated analogue, required for X-ray scattering, was implemented in a transmembrane peptide motif via manual solid-phase peptide synthesis (SPPS) following the fluorenylmethyloxycarbonyl (Fmoc)-strategy. The derived peptides were reconstituted in lipid vesicles as well as in highly aligned multilamellar lipid stacks and investigated via circular dichroism (CD) and X-ray reflectivity. Thereby, it has been revealed that the bulky iodine probe neither causes conformational change of the peptide structure nor lamellar disordering of the membrane complexes. PMID- 19565580 TI - Targeted 13C-13C distance measurements in a microcrystalline protein via J decoupled rotational resonance width measurements. AB - Rotational resonance width (R(2)W) magic-angle spinning (MAS) NMR experiments are performed to measure (13)C-(13)C distances in the hydrophobic core of the microcrystalline model protein G(Beta1). Such inter-residue distances are of particular value in NMR structure determinations. The experiments are done at a Larmor frequency of 750 MHz (1)H where the contribution of (13)C chemical shift anisotropy (CSA) to the R(2) transfer mechanism is significant. To minimize line broadening in the 2D spectra, we employ a combination of even/odd isotopic labeling with [1,3-(13)C] glycerol, and J-decoupling in the indirect dimension. This results in high-precision distance measurements between aromatic side chains of three tyrosine residues and distant methyl groups in the hydrophobic core of the protein. Even in the absence of information on the relative orientation of the shift tensors, we obtain relatively high precision data, which can be further improved by additional constraints on the tensor orientations. PMID- 19565581 TI - Microscopic and spectroscopic observations in the bio-nanoworld. AB - Biophysics in Linz: In February biophysicists from across the world converged on Linz for two biophysical conferences. The city which is the Cultural Capital of Europe 2009, provided the perfect environment for fruitful discussions on single molecule techniques in biophysics, bio-nanotechnology, cell biology, and drug discovery. PMID- 19565583 TI - Isocyanate- and isothiocyanate-derived Ru(IV)-based alkylidenes: synthesis, structure, and activity. AB - The synthesis and characterization of a series of isocyanate- and isothiocyanate derived second generation Grubbs-Hoveyda-type ruthenium-alkylidene complexes, that is, [Ru(N=C=O)(2)(IMesH(2))(=CH-2-(2-PrO)-C(6)H(4))] (1), [Ru(N=C=O)(2)(1,3 dimesityl-3,4,5,6-tetrahydropyrimidin-2-ylidene)(=CH-2-(2-PrO)-C(6)H(4))] (2), [Ru(N=C=S)(2)(IMesH(2))(=CH-2-(2-PrO)-C(6)H(4))] (3), and [Ru(N=C=S)(2)(1,3 dimesityl-3,4,5,6-tetrahydropyrimidin-2-ylidene)(=CH-2-(2-PrO)-C(6)H(4))] (4), and their activity in various metathesis reactions are described. Compounds 1-4 were prepared by reaction of the parent complexes [RuCl(2)(IMesH(2))(=CH-2-(2 PrO)C(6)H(4))] (5) (IMesH(2)=1,3-bis-(2,4,6-trimethylphenyl)-4,5-dihydroimidazol 2-ylidene) and [RuCl(2)(1,3-dimesityl-3,4,5,6-tetrahydropyrimidin-2-ylidene)(=CH 2-(2-PrO)-C(6)H(4))] (6) with silver cyanate and thiocyanate, respectively. The X ray structure of 1 was determined, confirming the isocyanate-type bonding of the ligand. The isothiocyanate-type bonding in 3 and 4 was unambiguously confirmed by IR and (13)C NMR spectroscopy. The isocyanate-derived complexes 1 and 2 were found to be excellent catalysts for the ring-opening metathesis polymerization (ROMP) of cis-cycloocta-1,5-diene (COD). Both 1 and 2 yielded poly(COD) with a trans-content of about 80 %. First-order kinetics with unprecedentedly high rate constants of polymerization (k(p)=0.068 and 0.26 s(-1), respectively) were observed. Compounds 3 and 4 were also active initiators for the ROMP of COD, however, they generated poly(COD) with a cis-content of 80 and 67 %, respectively. Complexes 1 and 2 also showed good catalytic activity in cross metathesis (CM) reactions. Finally, 1-4 were also found to be excellent catalysts for the regioselective cyclopolymerization of diethyl 2,2-dipropargylmalonate (DEDPM), resulting in poly(DEDPM) almost entirely based on five-membered repeat units, that is, cyclopent-1-ene-1,2-vinylenes. PMID- 19565584 TI - Soluble polymeric dual sensor for temperature and pH value. PMID- 19565585 TI - Rotational spectral signatures of four tautomers of guanine. PMID- 19565586 TI - Fluorinated quinine alkaloids: synthesis, X-ray structure analysis and antimalarial parasite chemotherapy. AB - Herein we report the synthesis of a series of C9-fluorinated quinine alkaloids by direct nucleophilic deoxyfluorination. This transformation gives rise to products bearing both S- and R-configured monofluoromethylene functionalities, consistent with an S(N)1-like mechanism. Furthermore, a series of ring-expanded 1 azabicyclo[3.2.2]nonane systems were generated by a skeletal rearrangement of the quinuclidine core. The modified alkaloids were converted to the corresponding hydrochloride salts and characterised by single-crystal X-ray diffraction analysis. The preference of the benzylic fluorine atom to adopt a gauche conformation relative to the protonated quinuclidine nitrogen atom was consistently observed throughout the cage-conserved compounds. Conversely, the molecular architecture of the 1-azabicyclo[3.2.2]nonane systems enforced an anti relationship between the fluorine atom and the protonated tertiary amine. This constitutes the first X-ray evidence of a vicinal fluorine atom at a stereogenic centre positioned anti to a substituted ammonium cation. The pharmacological efficacy of these compounds was assessed in vitro against the NF54 strain of Plasmodium falciparum (sensitive to all known antimalarial drugs). IC(50) values of as low as 267 nM were observed; this highlights the potential of these materials in developing novel agents for parasite chemotherapy. PMID- 19565587 TI - Microfluidic confinement of single cells of bacteria in small volumes initiates high-density behavior of quorum sensing and growth and reveals its variability. PMID- 19565588 TI - Efficient access to oseltamivir phosphate (Tamiflu) via the O-trimesylate of shikimic acid ethyl ester. PMID- 19565589 TI - Reversible 100% linkage isomerization in a single-crystal to single-crystal transformation: photocrystallographic identification of the metastable [Ni(dppe)(eta1-ONO)Cl] isomer. PMID- 19565591 TI - The ruthenium-catalyzed hydrovinylation of internal alkynes by acrylates: an atom economic approach to highly substituted 1,3-dienes. PMID- 19565590 TI - Constructing and exploiting the fluorescent protein paintbox (Nobel Lecture). PMID- 19565592 TI - Transition-metal-catalyzed ring opening of hetero-Diels-Alder adducts. PMID- 19565593 TI - Selectivity of competitive multivalent interactions at interfaces. AB - The development of synthetic, low-molecular-weight ligand receptor systems for the selective control of biomolecular interactions remains a major challenge. Binding of oligohistidine peptides to chelators containing Ni2+-loaded nitrilotriacetic acid (NTA) moieties is one of the most widely used and best characterised recognition systems. Recognition units containing multiple NTA moieties (multivalent chelator headgroups, MCHs) recognise oligohistidines with substantially increased binding affinities. Different multivalencies both at the level of the MCH and at that of the oligohistidine ligand provide a powerful means to vary the affinity of the interaction systematically. Here we have explored the selectivity for the binding of different oligohistidines to immobilised MCH. Using microarrays of mono-, bis-, tris- and tetrakis-NTA chelators spotted at different surface densities, we explored the ability of these binders to discriminate fluorescently labelled hexa- and decahistidine peptides. When hexa- and decahistidine were tested alone, the discrimination of ligands showed little dependence either on the nature or on the density of the chelator. In contrast, coincubation of both peptides decreased the affinity of hexahistidine, increased the affinity of decahistidine, and made the binding of decahistidine highly dependent on MCH density. Kinetic binding assays by dual colour total internal reflection fluorescence spectroscopy revealed active exchange of His6 by His10 and confirmed the high selectivity towards His10. Our results establish the key role of surface multivalency for the selectivity of multivalent interactions at interfaces. PMID- 19565594 TI - Solid-state NMR studies of adenosine 5'-triphosphate freeze-trapped in the nucleotide site of Na,K-ATPase. PMID- 19565595 TI - Selective detection of phosphotyrosine in the presence of various phosphate containing biomolecules with the aid of a terbium(III) complex. PMID- 19565596 TI - Multivalent manno-glyconanoparticles inhibit DC-SIGN-mediated HIV-1 trans infection of human T cells. PMID- 19565597 TI - Very fast product release and catalytic turnover of DNA photolyase. PMID- 19565598 TI - Pharmacophore-based discovery of small-molecule inhibitors of protein-protein interactions between HIV-1 integrase and cellular cofactor LEDGF/p75. AB - The cellular protein lens epithelium-derived growth factor, or transcriptional coactivator p75 (LEDGF/p75), plays a crucial role in HIV integration. The protein protein interactions (PPIs) between HIV-1 integrase (IN) and its cellular cofactor LEDGF/p75 may therefore serve as targets for the development of new anti HIV drugs. In this work, a structure-based pharmacophore model for potential small-molecule inhibitors of HIV-1 IN-LEDGF/p75 interaction was developed using the LigandScout software. The 3D model obtained was used for virtual screening of our in-house chemical database, CHIME, leading to the identification of compound CHIBA-3002 as an interesting hit for further optimization. The rational design, synthesis and biological evaluation of four derivatives were then carried out. Our studies resulted in the discovery of a new and more potent small molecule (7, CHIBA-3003) that is able to interfere with the HIV-1 IN-LEDGF/p75 interaction at micromolar concentration, representing one of the first compounds to show activity against these specific PPIs. Docking simulations were subsequently performed in order to investigate the possible binding mode of our new lead compound to HIV-1 IN. This study is a valid starting point for the identification of anti-HIV agents with a different mechanism of action from currently available antiviral drugs. PMID- 19565599 TI - Synthesis of dimeric quinazolin-2-one, 1,4-benzodiazepin-2-one, and isoalloxazine compounds as inhibitors of amyloid peptides association. AB - The synthesis of dimeric compounds derived from quinazolin-2-one and 1,4 benzodiazepin-2-one possessing a piperazine or homopiperazine spacer was investigated. In addition, a piperazine spacered bis-isoalloxazine and a bis riboflavin compound were prepared and their ability to interrupt the association of prion proteins and Alzheimer-specific Abeta peptides was investigated using a fast screening system based on flow cytometry. The bis-isoalloxazine 14 was identified as a new lead structure. PMID- 19565600 TI - Synthesis of novel 2,5-disubstituted 1,3,4-thiadiazoles for their potential anticonvulsant activity: pharmacophoric model studies. AB - A series of novel N(1)-[5-(4-substituted phenyl)-1,3,4-thiadiazol-2-yl]-N(4)-(4 substituted benzaldehyde)-semicarbazone 1-12, N(1)-[5-(4-substituted phenyl) 1,3,4-thiadiazol-2-yl]-N(4)-[1-(4-substituted phenyl)ethanone]-semicarbazone 13 16, and N(1)-[5-(4-substituted phenyl)-1,3,4-thiadiazol-2-yl]-N(4)-[1-(4 substituted phenyl) (phenyl) methanone]-semicarbazone 17-20 were synthesized for their anticonvulsant activity. The chemical structures of the compounds were proved by elemental and spectral (IR, (1)H-NMR,( 13)C-NMR, and MS) analysis. The anticonvulsant potential of the compounds was investigated using maximal electroshock seizure (MES) and subcutaneous pentylenetrtrazole (scPTZ) models. Compound 19 was found to possess significant anticonvulsant activity in both the models employed for anticonvulsant evaluation. Compounds 8, 13, 15, and 16 also demonstrated a marked anticonvulsant property. The results of the present study validated that the pharmacophore model with four binding sites is essential for anticonvulsant activity. The efforts were also made to establish structure activity relationships among the synthesized compounds. PMID- 19565601 TI - Multi-component synthesis of dihydropyrimidines by iodine catalyst at ambient temperature and in-vitro antimycobacterial activity. AB - An efficient and simple three-component domino synthesis of some new dihydropyrimidines (DHPMs) from aromatic aldehydes, 1,3-dicarbonyl compounds and N-(3-chloro-4-fluorophenyl)urea using molecular iodine as catalyst is described. The 1-substituted dihydropyrimidines were isolated in good to excellent yields (78-90%) within a short reaction time (4-6 h) at ambient temperature. The biological evaluation revealed that the newly synthesized compounds (4a-i and 5a i) exhibited moderate antimycobacterial activity against Mycobacterium tuberculosis H(37) RV. PMID- 19565602 TI - Synthesis and cytotoxic activity of novel amidine analogues of bis(2 chloroethyl)amine. AB - Novel nitrogen mustard agents 7-12 involving 4-(N,N-bis(2 chloroethyl)aminophenyl)propylamine linked to a 5-(4-N-alkylamidinophenyl)-2 furancarboxylic acid moiety by the formation of an amide bond have been synthesized, characterized, and evaluated for their in-vitro cytotoxic activity against MDA-MB-231 and MCF-7 human breast cancer cells. Evaluation of the cytotoxicity of 7-12 employing a MTT assay and inhibition of [(3)H]thymidine incorporation into DNA demonstrated that these compounds exhibit remarkable cytotoxic effects in comparison with 4-[bis(2-chloroethyl)amino]benzenebutanoic acid. Compounds 7 and 9, which possess a cationic amidine and 4,5-dihydro-1H imidazol function moiety are approximately ten times more potent than 4-[bis(2 chloroethyl)amino]benzenebutanoic acid. The new compounds were evaluated as DNA topoisomerase II inhibitors. The cytotoxicity of the compounds 7-12 correlates with their DNA-binding affinities and their relative potency as topoisomerase II inhibitors. PMID- 19565603 TI - Benzothiazole incorporated barbituric acid derivatives: synthesis and anticonvulsant screening. AB - A series of 1-(6-substituted-1,3-benzothiazol-2-yl)-3-(substituted phenyl)hexahydro-2,4,6-pyrimidinetriones 4a-t were synthesized starting from substituted anilines. These compounds contained two active anticonvulsant pharmacophores, benzothiazole and barbituric acid. Structures of the compounds were confirmed on the basis of different spectroscopic techniques. All the compounds were evaluated for their anticonvulsant activity. Three compounds 4c, 4d, and 4s showed promising anticonvulsant activities in Maximal Electroshock Seizure test (MES) and subcutaneous pentylenetetrazole test (scPTZ). They also displayed a wide safety profile when tested for the minimal motor impairment test. PMID- 19565608 TI - Prenatal 2D and 3D ultrasound diagnosis of diprosopus: case report with post mortem magnetic resonance images (MRI) and review of the literature. PMID- 19565609 TI - Maternal plasma pentraxin 3 at 11 to 13 weeks of gestation in hypertensive disorders of pregnancy. AB - OBJECTIVE: To investigate whether in pregnancies that subsequently develop preeclampsia (PE), the maternal plasma concentration of the inflammatory factor pentraxin 3 (PTX3) at 11-13 weeks of gestation is increased and whether such a possible increase is associated with uterine artery pulsatility index (PI). METHODS: The concentration of plasma PTX3 at 11-13 weeks was measured in a case control study from 120 pregnancies that developed PE, including 27 who required delivery before 34 weeks (early PE), 87 cases of gestational hypertension (GH) and 207 normal controls. The median PTX3 multiple of the median (MoM) in the control and hypertensive groups were compared. Regression analysis was used to determine the significance of the association between plasma PTX3 and uterine artery PI. RESULTS: Plasma PTX3 was significantly higher in the early PE group (1.44 MoM; p < 0.0083) but not in late PE (1.11 MoM) or GH (1.10 MoM) compared to the controls (0.97 MoM). There was no significant association between plasma PTX3 levels and uterine artery PI in either the PE group (p = 0.693) or in the controls (p = 0.209). CONCLUSION: Increase in maternal plasma PTX3 in pregnancies that subsequently develop early PE is evident from 11-13 weeks but the underlying mechanism for such an increase remains uncertain. PMID- 19565613 TI - Addressable DNA-myoglobin photocatalysis. AB - Photoactivatable myoglobin containing a DNA oligonucleotide as a structural anchor was designed by using the reconstitution of artificial heme moieties containing Ru(3+) ions. This semisynthetic DNA-enzyme conjugate was successfully used for the oxidation of peroxidase substrates by using visible light instead of H(2)O(2) for the activation. The DNA anchor was utilized for the immobilization of the enzyme on the surface of magnetic microbeads. Enzyme activity measurements not only indicated undisturbed biofunctionality of the tethered DNA but also enabled magnetic separation-based enrichment and recycling of the photoactivatable biocatalyst. PMID- 19565615 TI - Cell fate transitions during stomatal development. AB - Stomata, the most influential components in gas exchange with the atmosphere, represent a revealing system for studying cell fate determination. Studies in Arabidopsis thaliana have demonstrated that many of the components, functioning in a signaling cascade, guide numerous cell fate transitions that occur during stomatal development. The signaling cascade is initiated at the cell surface through the activation of the membrane receptors TOO MANY MOUTHS (TMM) and/or ERECTA (ER) family members by the secretory peptide EPIDERMAL PATTERNING FACTOR1 (EPF1) and/or a substrate processed proteolytically by the subtilase STOMATAL DENSITY AND DISTRIBUTION1 (SDD1) and transduced through cytoplasmic MAP kinases (YODA (YDA), MKK4/MKK5, and MPK3/MPK6) towards the nucleus. In the nucleus, these MAP kinases regulate the activity of the basic helix-loop-helix (bHLH) proteins SPEECHLESS (SPCH), MUTE, and FAMA, which act in concert with the bHLH-Leu zipper protein SCREAM (SCRM) (and/or its closely related paralog, SCREAM2). This article reviews current insights into the role of this signaling cascade during stomatal development. PMID- 19565616 TI - How does graphene grow? Easy access to well-ordered graphene films. AB - The selective formation of large-scale graphene layers on a Rh-YSZ-Si(111) multilayer substrate by a surface-induced chemical growth mechanism is investigated using low-energy electron diffraction, X-ray photoelectron spectroscopy, X-ray photoelectron diffraction, and scanning tunneling microscopy. It is shown that well-ordered graphene layers can be grown using simple and controllable procedures. In addition, temperature-dependent experiments provide insight into the details of the growth mechanisms. A comparison of different precursors shows that a mobile dicarbon species (e.g., C(2)H(2) or C(2)) acts as a common intermediate for graphene formation. These new approaches offer scalable methods for the large-scale production of high-quality graphene layers on silicon based multilayer substrates. PMID- 19565617 TI - Greatly improved blood compatibility by microscopic multiscale design of surface architectures. PMID- 19565622 TI - Vitamin D in diabetes mellitus-a new field of knowledge poised for D-velopment. AB - This commentary reviews the current state of knowledge regarding the role of vitamin D in the pathogenesis of diabetes mellitus. In type 1 diabetes mellitus or in adult onset latent autoimmune diabetes (LADA), vitamin D exhibits immunomodulatory actions, influencing the activity of lymphocytes and interleukins. In type 2 diabetes mellitus vitamin D appears to act through different mechanisms, affecting insulin secretion and insulin sensitivity through its effects on the beta cells, mediators of inflammation and parathyroid hormone. Much work remains to be done in this new field of knowledge before the role of vitamin D in the pathogenesis of diabetes mellitus is completely understood. PMID- 19565618 TI - Fetal trisomy 13 and 21 mosaicism diagnosed at amniocentesis: a case report. PMID- 19565625 TI - NICE goes off the rails. PMID- 19565623 TI - Chemically modified porous silicon for laser desorption/ionization mass spectrometry of ionic dyes. AB - Desorption/ionization on silicon (DIOS) mass spectra of model ionic dyes methylene blue (MB+Cl-) and methyl orange (Na+MO-) were studied using p+ type derived porous silicon (PS) free layers. As-prepared PS (PS-H), the PS thermally oxidized at 300 degrees C (PS-OX), PS with chemically grafted cation-exchanging alkylsulfonic acid (PS-SO(3)H) and anion-exchanging propyl octadecyldimethylammonium chloride (PS-ODMA+Cl-) groups was tested as ionization platforms. Two mechanisms of the methylene blue desorption/ionization were found: (1) the formation of [MB + H]+* ion due to the reduction/protonation of MB+, which is predominant for PS-H and PS-OX platforms and (2) direct thermal desorption of the MB+ cation, prevailing for PS-SO3H. The fragmentation of the cation is significantly suppressed in the latter case. The samples of PS-SO3H and PS-ODMA+ Cl- efficiently adsorb the dyes of the opposite charge from their solutions via the ion-exchange. Consequent DIOS MS studies allow to detect only low fragmented ions (MB+ and MO-, respectively), demonstrating the potential of the ion-exchange adsorption combined with DIOS MS for the analysis of ionic organic compounds in solutions. PMID- 19565626 TI - Gap junctions are involved in cell migration in the early postnatal subventricular zone. AB - The massive migration of neuroblasts and young neurons through the anterior extension of the postnatal subventricular zone (SVZ), known as the rostral migratory stream (RMS) is still poorly understood on its molecular basis. In this work, we investigated the involvement of gap junctional communication (GJC) in the robust centrifugal migration from SVZ/RMS explants obtained from early postnatal (P4) rats. Cells were dye-coupled in homocellular and heterocellular pairings and expressed at least two connexins, Cx 43 and 45. Treatment with the uncoupler agent carbenoxolone (CBX, 10-100 microM) reversibly reduced outgrowth from SVZ explants, while its inactive analog, glycyrhizinic acid (GZA), had no effect. Consistent with a direct effect on cell migration, time-lapse video microscopy show that different pharmacological uncouplers cause an abrupt and reversible arrest of cell movement in explants. Our results indicate that GJC is positively involved in the migration of neuroblasts within the SVZ/RMS. PMID- 19565627 TI - Paternal deprivation induces dendritic and synaptic changes and hemispheric asymmetry of pyramidal neurons in the somatosensory cortex. AB - Similar to maternal care, paternal care is a source of neonatal sensory stimulation, which in primates and rodents has been shown to be essential for developing structure and function of sensory cortices. The aim of our study in the biparental rodent Octodon degus was to assess the impact of paternal deprivation on dendritic and synaptic development in the somatosensory cortex. We (i) quantified the amount of paternal care in relation to total parental investment and (ii) compared dendritic and synaptic development of pyramidal neurons in the somatosensory cortex of animals raised by a single mother or by both parents. On the behavioral level we show that paternal care comprises 37% of total parent-offspring interactions, and that the somatosensory stimulation provided by the fathers primarily consists of huddling, licking/grooming, and playing. On the morphological level we found that, compared with offspring raised by both parents (mother and father), the father-deprived animals displayed significantly reduced spine numbers on the basal dendrites of pyramidal neurons. Furthermore, paternal deprivation induces hemispheric asymmetry of the dendritic morphology of somatosensory pyramidal neurons. Father-deprived animals show shorter and less complex basal dendrites in the left somatosensory cortex compared with the right hemisphere. These findings indicate that paternal deprivation results in delayed or retarded dendritic and synaptic development of somatosensory circuits. PMID- 19565628 TI - Oscillating migration and the epidemics of silicosis, tuberculosis, and HIV infection in South African gold miners. AB - BACKGROUND: Hundreds of thousands of men from rural areas of South Africa and neighboring countries have come to seek work in the gold mines. They are not immigrants in the usual sense as they work for periods in the mines, go home, and then return. This is termed oscillating or circular migration. Today we have serious interrelated epidemics of silicosis, tuberculosis, and HIV infection in the gold mining industry. METHODS: This article discusses the role of oscillating migration in fuelling these epidemics, by examining the historical, political, social, and economic contexts of these diseases. RESULTS: The impact of silicosis, tuberculosis, and HIV infection extends beyond individual miners to their families and communities. CONCLUSION: Failure to control dust and tuberculosis has resulted in serious consequences decades later. The economic and political migrant labor system provided the foundations for the epidemics seen in southern Africa today. PMID- 19565629 TI - Declining health insurance access among US Hispanic workers: not all jobs are created equal. AB - INTRODUCTION: Approximately 18% of the U.S. population are uninsured, a proportion that may continue to rise, particularly among Hispanics, as the cost of medical care increases faster than the growth in wages. METHODS: Health insurance trends were analyzed by race-ethnic category, and among Hispanic workers by occupation type and industrial sector, using data on employed respondents > or =18 years from 1997 to 2007 National Health Interview Survey (NHIS) (mean annual n = 17,392, representing 123 million US workers on average over this 11 year period). RESULTS: From 1997 to 2007, the relative decline in health insurance coverage for US workers was greatest among Hispanics (7.0%). Hispanic workers in the Construction and Services industries had the greatest overall decline in coverage (24.9% and 14.7%), as well as Hispanic blue collar workers (14.0%). CONCLUSION: Hispanic workers in general, and those employed in blue collar, construction, and services sectors in particular, are at greater risk for poor access to health care due to a lack of health insurance coverage. PMID- 19565630 TI - Increased expression of hyaluronic acid binding protein 1 is correlated with poor prognosis in patients with breast cancer. AB - BACKGROUND AND OBJECTIVES: Hyaluronic acid binding protein 1 (HABP1), a family of proteins interacting with hyaluronan (HA), had been associated with cell adhesion and tumor invasion. The aim of this study was to investigate the correlation between clinicopathologic factors and patient survival time with the expression of HABP1 in breast cancer patients. METHODS: Expression of HABP1 mRNA and protein were detected with real-time quantitative PCR and immunohistochemical staining in 63 breast cancer and non-cancerous matched tissues. RESULTS: The mRNA expression level of HABP1 was unrelated to the patient's age, tumor size, histological grade, TNM stage. However, it proved to be positively related to axillary nodes metastasis (P = 0.008). Furthermore, it was shown that the survival rate of patients with low HABP1 expression was significantly higher than that of patients with high HABP1 expression (P = 0.025). Multivariate analysis revealed that HABP1 mRNA expression level was a significant factor for predicting prognosis (P = 0.022). The immunohistochemistry results showed that the expression level of HABP1 in breast cancer cells was higher than that in normal breast cells. CONCLUSION: HABP1 might be an independent predictive factor for breast cancer prognosis and up-regulation of HABP1 might play an important role in the metastasis of breast cancer. PMID- 19565631 TI - Effect of the substrate and catalyst chirality on the diastereoselective epoxidation of limonene using Jacobsen-type catalysts. AB - Chiral and achiral Jacobsen's catalysts in their homogeneous form or immobilized on Al-MCM-41 exhibit similar catalytic activity during diastereoselective epoxidation of limonene when in situ generated dimethyldioxirane is used as oxidizing agent. Experimental observations suggest that not only the catalyst chiral center but also the substrate chiral center participates in the preferential formation of most diastereomers. Remarkable turnover numbers (TON), up to 288, was achieved over the heterogeneous catalysts in comparison to their homogeneous counterparts (TON up to 46). Catalyst leaching rather than catalyst oxidative degradation was identified as the main source of catalyst deactivation during reutilization tests. PMID- 19565633 TI - Toxicity of parathion on embryo and yolk-sac larvae of gilthead seabream (Sparus aurata l.): effects on survival, cholinesterase, and carboxylesterase activity. AB - This study was conducted to examine the acute toxicity of the organophosphorus pesticide (OP) parathion on embryos and yolk-sac larvae of gilthead seabream (Sparus aurata), and to investigate the effects of this compound on cholinesterase and carboxylesterase activity of seabream larvae in the phase of endogenous feeding. The 72-h LC50 for yolk-sac larvae (0.523 mg L-1) was about two-fold lower than the 48-h LC50 for embryos (1.005 mg L-1). Parathion significantly inhibited the activity of ChE and CaE activity in yolk sac larvae but there were not significant differences in the sensitivity of both esterases to parathion as inferred by their 72-h IC50 values. Larvae exposed to parathion for 72 h showed a 70% inhibition of the whole body acetylcholinesterase at approximately the LC50. PMID- 19565632 TI - Tolerance of Silene vulgaris to copper: population-related comparison of selected physiological parameters. AB - Tolerance of three Slovak populations of Silene vulgaris [from the localities "Harmanec" (total soil Cu 34.1 mg kg-1), "Lubietova"; (total soil Cu 831.6 mg kg 1) and "Spania dolina" (total soil Cu 1368.7 mg kg-1) referred as SV1, SV2, and SV3, respectively] and Silene dioica (SD; from the locality "Spania dolina") exposed to 60 MUM Cu for 7 days has been studied. Respective controls with 0.21 MUM Cu were also cultured. SV3 and SD contained higher total shoot Cu than SV1. Methanol-soluble shoot Cu represented 60, 59, 59, and 55% from total shoot Cu and roots contained 16.7-, 20.4-, 9.9- and 28.9-fold more Cu than shoots in SV1, SV2, SV3, and SD, respectively. Soluble proteins were the least affected in SV3 and SD when Cu-exposed and control plants were compared. Root hydrogen peroxide and superoxide showed similar trend and were correlated with enhancement of ascorbate and guaiacol-peroxidase activities. Malondialdehyde accumulation increased in SV1 and SV2. Phenylalanine ammonia-lyase activity and total soluble phenols were higher in SV3 Cu-exposed plants compared to SV1 and SV2. Shikimate dehydrogenase activity was enhanced in all Cu-exposed populations. Cinnamyl alcohol dehydrogenase activity and root lignin content were not affected and polyphenol oxidase activity was not detected. Within 14 detected free amino acids, majority of them decreased preferentially in the roots. Only serine increased in shoots and decreased in roots of all populations. Potassium content was not affected in SV3. Na, Ca, and Fe increased in roots of Cu-exposed plants of all populations, while Mg was the least affected and Zn increased in SV2 and SV3 shoots and roots. This study revealed the highest tolerance in population from the locality with the highest soil Cu content. Present findings as complex metabolic responses to Cu stress with special emphasis on phenolic metabolism are discussed. PMID- 19565634 TI - Platelet-activating factor-stimulated production of reactive oxygen species in ovarian granulosa cells from periovulatory follicles. AB - The platelet-activating factor (PAF) is a proinflammatory lipid present in the fluid of ovarian Graafian follicle. Ovarian blockage of the PAF receptor (PAFr) reduces ovulations in the rat whereas underlying mechanism is poorly understood. Mural granulosa cells (MGC) were mechanically isolated from the theca interna of bovine periovulatory follicle. The mRNA abundance for PAFr, progesterone receptor and cyclooxygenase-2 were measured by real-time PCR. Cytosolic calcium (Ca2+) concentration was assayed by microscopy using Fura-2 AM as indicator, 8 isoprostaglandin F(2alpha) (8-isoPGF(2alpha)) by an ELISA kit. Fluorescent products arising from intracellular oxidation of hydroethidine (HE) and dihydrorhodamine (DHR) were quantified by flow cytometry. The cells expressed PAFr mRNA and PAFr protein and responded to cPAF (nonhydrolyzable form of PAF) with a pulsating increase in Ca2+, demonstrating functional PAFr. Elevation of Ca2+ was reversed by WEB-2086, an inverse PAFr agonist. cPAF elevated the level of 8-isoPGF(2alpha) in the medium of MGC cultured with luteinizing hormone (LH). cPAF alone had no significant influence on the oxidation of HE and DHR, or 8 isoPGF(2alpha) level. In MGC from vital periovulatory follicle, PAF and LH signaling plays an important role in regulating the production of excessive oxidants. Blockage of PAFr seems to interfere with these regulatory processes essential for ovulation. PMID- 19565635 TI - A simple method for multiday imaging of slice cultures. AB - The organotypic slice culture (Stoppini et al. A simple method for organotypic cultures of nervous tissue. 1991;37:173-182) has become the method of choice to answer a variety of questions in neuroscience. For many experiments, however, it would be beneficial to image or manipulate a slice culture repeatedly, for example, over the course of many days. We prepared organotypic slice cultures of the auditory brainstem of P3 and P4 mice and kept them in vitro for up to 4 weeks. Single cells in the auditory brainstem were transfected with plasmids expressing fluorescent proteins by way of electroporation (Haas et al. Single cell electroporation for gene transfer in vivo. 2001;29:583-591). The culture was then placed in a chamber perfused with oxygenated ACSF and the labeled cell imaged with an inverted wide-field microscope repeatedly for multiple days, recording several time-points per day, before returning the slice to the incubator. We describe a simple method to image a slice culture preparation during the course of multiple days and over many continuous hours, without noticeable damage to the tissue or photobleaching. Our method uses a simple, inexpensive custom-built insulator constructed around the microscope to maintain controlled temperature and uses a perfusion chamber as used for in vitro slice recordings. PMID- 19565636 TI - Changes in MAPK pathway in neonatal and adult testis following fetal estrogen exposure and effects on rat testicular cells. AB - Concerns have been raised about the possible role of the phytoestrogen genistein and the xenoestrogen Bisphenol A (BPA) as endocrine disruptors. In the present study, we examined the effects of fetal exposure to genistein and BPA on the Mitogen-activated protein kinase (MAPK) pathway and on testicular cell populations in neonatal and adult rat testes. At postnatal day (PND) 3, genistein (0.1-10 mg/kg/day) and BPA (1-200 mg/kg/day) induced Raf1 and Erk1/2 mRNA and protein increases in testes, mainly in Sertoli cells. No changes were seen for Mek1. At PND60, Erk1/2 protein expression remained robust in Sertoli cells and in some spermatogonia. Raf1 was predominant in Leydig cells while Mek1 was expressed strongly in spermatogonia, and they were both expressed in pachytene spermatocytes. No consistent change was seen in these proteins at PND60. Transient effects were observed on germ cell populations, while the only remaining effect on adult testicular cells was an increase in Leydig cell number. Rats exposed in utero to the two compounds did not present significant changes in circulating testosterone levels, suggesting normally functioning adult Leydig cells. Furthermore, Sertoli cell numbers were not affected by exposure to genistein and BPA. Finally, around 10% of genistein and BPA exposed rats were sterile, whereas all control rats were fertile. These data suggest that fetal exposure to genistein and BPA exerts transient effects in rat testes and that the changes observed at PND3 did not correlate with relevant changes in germ cell populations, Leydig cell function, or fertility in the adult. PMID- 19565637 TI - Simultaneous flow cytometric quantification of plant nuclear DNA contents over the full range of described angiosperm 2C values. AB - Flow cytometry provides a rapid, accurate, and simple means to determine nuclear DNA contents (C-value) within plant homogenates. This parameter is extremely useful in a number of applications in basic and applied plant biology; for example, it provides an important starting point for projects involving whole genome sequencing, it facilitates characterization of plant species within natural and agricultural settings, it allows facile identification of engineered plants that are euploid or that represent desired ploidy classes, it points toward studies concerning the role of C-value in plant growth and development and in response to the environment and in terms of evolutionary fitness, and, in uncovering new and unexpected phenomena (for example endoreduplication), it uncovers new avenues of scientific enquiry. Despite the ease of the method, C values have been determined for only around 2% of the described angiosperm (flowering plant) species. Within this small subset, one of the most remarkable observations is the range of 2C values, which spans at least two orders of magnitude. In determining C-values for new species, technical issues are encountered which relate both to requirement for a method that can provide accurate measurements across this extended dynamic range, and that can accommodate the large amounts of debris which accompanies flow measurements of plant homogenates. In this study, the use of the Accuri C6 flow cytometer for the analysis of plant C-values is described. This work indicates that the unusually large dynamic range of the C6, a design feature, coupled to the linearity of fluorescence emission conferred by staining of nuclei using propidium iodide, allows simultaneous analysis of species whose C-values span that of almost the entire described angiosperms. PMID- 19565638 TI - Disease-specific biomarker discovery by aptamers. AB - RNA and DNA aptamers developed by an in vitro selection process, Systematic Evolution of Ligands by EXponential enrichment (SELEX), comprise a novel class of high-affinity and specific capture agents, which can be easily modified for cytometry and in vivo applications. A novel application of this technique (Cell SELEX) explores the expression of cell surface epitopes that differ between two given cell types or between healthy and diseased cells. Using whole cells as targets, aptamer libraries can be identified that bind to biomarkers expressed by target cells and not by any other cells. Aptamers have been developed that specifically interact with cell surface epitopes of trypanosomes or distinguish between the differences in molecular signature of somatic and cancer cells. Aside from its use for target cell identification by image and flow cytometry and laser scanning microscopy, aptamers can be used for ligand-mediated purification and identification of their binding proteins in target cell membranes. In this review, we discuss an approach for the development of aptamers targeting parasite derived surface proteins of Trypanosoma and Plasmodium. PMID- 19565639 TI - Human angiomotin-like 1 associates with an angiomotin protein complex through its coiled-coil domain and induces the remodeling of the actin cytoskeleton. AB - Angiostatin is a potent inhibitor of angiogenesis. One mechanism through which angiostatin inhibits angiogenesis is by binding to the cell surface protein p80 angiomotin. The p80-angiomotin protein promotes angiogenesis, in part, by conferring a hypermigratory phenotype to endothelial cells. Although p80 angiomotin is extensively characterized, less is known about the related protein angiomotin-like 1. We report that angiomotin-like 1 forms part of a protein complex containing p80-angiomotin. Structure-function studies revealed that angiomotin-like 1 associates with this p80-angiomotin-containing complex via its coiled-coil domain. Since p80-angiomotin plays a role in cell migration, a process that involves the remodeling of the actin cytoskeleton, we then addressed the hypothesis that angiomotin-like 1 may interact with the cytoskeleton. Immunofluorescence studies reveal that angiomotin-like 1 not only co-localizes with filamentous actin but also significantly modifies the architecture of the actin cytoskeleton. Regarding migration, angiomotin-like 1 increases the velocity of migration and decreases the persistence of migration directionality. Together these observations strongly suggest that angiomotin-like 1 is involved in actin cytoskeleton-based processes, in part, via its interaction with a p80-angiomotin containing complex and the actin cytoskeleton. These findings have important implications for angiogenesis-driven disease since angiomotin and angiomotin-like 1 are both expressed in capillaries. PMID- 19565640 TI - NANOS3 gene mutations in men with isolated sterility phenotype. PMID- 19565642 TI - Ion current activity and molecules modulating maturation and growth stages of ascidian (Ciona intestinalis) oocytes. AB - Electrophysiological techniques were used to study ion currents in the ascidian Ciona intestinalis oocyte plasma membranes during different stages of growth and meiosis. Three stages (A, B, C) of immature oocytes were discriminated in the ovary, with the germinal vesicle (GV) showing specific different features of growth and maturation. Stage A (pre-vitellogenic) oocytes exhibited the highest L type Ca(2+)current activity, and were incompetent for meiosis resumption. Stage B (vitellogenic) oocytes showed Na(+) currents that remained high during the maturation, up to the post-vitellogenic stage C oocytes. The latter had acquired meiotic competence, undergoing spontaneous maturation and interacting with the spermatozoon. However, fertilized oocytes did not produce normal larvae, suggesting that cytoplasmic maturation plays a specific role in embryo development. Spontaneous maturation was inhibited at low pH whereas trypsin was able to trigger germinal vesicle breakdown (GVBD) regardless of pH; in addition spontaneous maturation was not affected by removal of follicle cells or by inhibiting junctional communication between oocyte and follicle cells. Taken together these results imply: (i) Ca(2+) and Na(+) currents are involved in meiotic progression, growth, and acquisition of meiotic competence; (ii) trypsin like molecules may have a role as candidates for providing the physiological stimulus to resume meiosis. Finally, we provide evidence that follicle cells in Ciona are not involved in triggering GVBD as it occurs in other ascidians. PMID- 19565641 TI - Aurora kinase B modulates chromosome alignment in mouse oocytes. AB - The elevated incidence of aneuploidy in human oocytes warrants study of the molecular mechanisms regulating proper chromosome segregation. The Aurora kinases are a well-conserved family of serine/threonine kinases that are involved in proper chromosome segregation during mitosis and meiosis. Here we report the expression and localization of all three Aurora kinase homologs, AURKA, AURKB, and AURKC, during meiotic maturation of mouse oocytes. AURKA, the most abundantly expressed homolog, localizes to the spindle poles during meiosis I (MI) and meiosis II (MII), whereas AURKB is concentrated at kinetochores, specifically at metaphase of MI (Met I). The germ cell-specific homolog, AURKC, is found along the entire length of chromosomes during both meiotic divisions. Maturing oocytes in the presence of the small molecule pan-Aurora kinase inhibitor, ZM447439 results in defects in meiotic progression and chromosome alignment at both Met I and Met II. Over-expression of AURKB, but not AURKA or AURKC, rescues the chromosome alignment defect suggesting that AURKB is the primary Aurora kinase responsible for regulating chromosome dynamics during meiosis in mouse oocytes. PMID- 19565644 TI - HongrES1, a cauda epididymis-specific protein, is involved in capacitation of guinea pig sperm. AB - Capacitation requires removal of proteins secreted by the cauda epididymis. Previously, we isolated and cloned the HongrES1 gene from rat cauda epididymis and found that it was exclusively expressed there. Here we report that HongrES1 mRNA is also expressed in the guinea pig cauda epididymis using Northern blot analysis, and the molecular weight of its cognate protein is approximately 48 kDa by Western blot analysis. Therefore, we investigated whether HongrES1 was involved in regulation of sperm capacitation in guinea pig. The results show that HongrES1 antisera (HA) significantly enhances sperm capacitation with maximal stimulation at a dilution of 1:500. Capacitation was reversed when capacitated spermatozoa were re-exposed to HongrES1 protein (HP, 0.25 microg/ml). In other words, HP acted as a decapacitation factor. HA accelerated the onset of capacitation and promoted a sperm hyperactivated motility response. Sperm capacitation was accelerated by HA stimulation of extracellular calcium influx while HP prevented extracellular calcium from influxing. Indirect immunofluorescence staining finds HP localized over the acrosomal anterior region of the sperm head, which exfoliates gradually during capacitation incubation, and completely disappeared after the acrosome reaction. Thus, HongrES1 expressed by the cauda epididymis is a novel molecule that regulates the physiology of guinea pig sperm prior to fertilization. PMID- 19565645 TI - The role of antiphospholipid antibodies toward the protein C/protein S system in venous thromboembolic disease. AB - The association between venous thromboembolism (VTE) and antibodies anti-Protein C (PC)/Protein S (PS) is still uncertain. We performed a case-control study to determine the risk of VTE related to the presence of these auto-antibodies considered independently of the presence of lupus anticoagulant (LAC) or anti cardiolipin antibodies (ACA). One hundred thirty-five patients with idiopathic VTE and 164 healthy subjects were enrolled. Anti-PC and anti-PS antibodies (both IgG and IgM) were assessed using commercially available ELISA kits. Among cases there was a higher prevalence of elevated anti-PC IgM antibodies than in controls (OR 2.44, 95%CI 1.00-5.94). The presence of anti-PC IgG and anti-PS IgG and IgM antibodies was also higher in cases than in controls, but the difference was not statistically significant. Only five patients had both anti-PC or anti-PS antibodies and LAC or ACA. We performed a stepwise multivariate logistic regression analysis showing that anti-PC IgM>958 percentile was a significant predictor of VTE after adjustment for LAC or ACA (OR 2.52, 95%CI 1.01-6.24)). Larger prospective studies are needed to confirm this finding. PMID- 19565643 TI - Diverse functions of HBEGF during pregnancy. AB - The establishment of pregnancy requires an intimate physical interaction and a molecular dialogue between the conceptus and the maternal reproductive tract that commences at implantation and continues until the placenta is formed and fully functional. Failure of the regulatory processes that ensure the fidelity of this relationship can precipitate a catastrophic pregnancy loss. One of the earliest identified molecular mediators of blastocyst implantation is heparin-binding epidermal growth factor (EGF)-like growth factor (HBEGF), which signals between the endometrium and implanting trophoblast cells to synchronize their corresponding developmental programs. HBEGF expression by trophoblast cells of the developing placenta appears to regulate extravillous differentiation and provide cytoprotection in a sometimes-hostile environment. This versatile member of the EGF signaling system will be examined in light of its associations with key events during early pregnancy. PMID- 19565646 TI - Acute weight gain and diastolic dysfunction as a potent risk complex for post stem cell transplant atrial fibrillation. AB - The management of atrial fibrillation (AF) following stem cell transplant (SCTX) is often challenging because of the universal presence of profound bone marrow suppression. The incidence of and risk factors for AF/flutter following SCTX are not well known. A total of 395 multiple myeloma (MM) patients consecutively underwent SCTX between 2002 and 2005 at the Mayo Clinic, and 383 of whom, mean age 57 +/- 9 years, had no history of evidence of AF/flutter constituted the study population. During 1,002 person-years of follow up, 39 (10%) patients developed first AF/flutter (incidence of 39 per 1,000 person years), and 28 of these (72%) occurred within 21 days of SCTX. In multivariable-adjusted analyses, weight gain of > or = 7% in the 1st week post-SCTX (HR 3.68; P = 0.0120) and presence of diastolic dysfunction at MM diagnosis (HR 2.294; P = 0.0082) were independent predictors of AF/flutter. The risk of AF/flutter post-SCTX increased by about ninefold when both factors were present. Compared to age and sex-matched MM patients without SCTX, the risk of AF/flutter differed significantly only over the 1st year after MM diagnosis, during which SCTX was performed for the majority. Beyond the 1st year, there was no significant difference in risk of AF/flutter between the two groups. The data suggested that SCTX was associated with significantly increased risk of first AF/flutter, which typically occurred within the first 21 days of the transplant. Weight gain of > or = 7% was strongly predictive of first AF/flutter, and the risk was augmented by the presence of diastolic dysfunction at baseline. PMID- 19565647 TI - Pre-, intra-, and postoperative sonography of the abdominal wall in patients with incisional hernias repaired via a three-layered operative suture method. AB - We illustrate the various sonographic (US) appearances of the abdominal wall following this type of repair, including partial and complete recurrences. Correlation is made with CT imaging. The three-layered anatomical reconstruction of an incisional hernia is described. PMID- 19565648 TI - Iron overload and chronically transfused patients: numbers, scales, and clinical research. PMID- 19565649 TI - Synergistic antitumor effects of lenalidomide and rituximab on mantle cell lymphoma in vitro and in vivo. AB - Rituximab (RTX), a chimeric anti-CD20 antibody, is associated with direct induction of apoptosis and antibody-dependent cell-mediated cytotoxicity (ADCC) with clinical efficacy in mantle cell lymphoma (MCL). Lenalidomide (LEN), a novel immunomodulatory agent, sensitizes tumor cells and enhances ADCC. Our study attempted to elucidate the mechanism of LEN-enhanced RTX-mediated cytotoxicity of MCL cells. We found that LEN and RTX induced growth inhibition of both cultured and fresh primary MCL cells. LEN enhanced RTX-induced apoptosis via upregulating phosphorylation of c-Jun N-terminal protein kinases (JNK), Bcl-2, Bad; increasing release of cytochrome-c; enhancing activation of caspase-3, -8, -9 and cleavage of PARP. Meanwhile, LEN activated NK cells and increased CD16 expression on CD56(low)CD16(+) NK cells. Whole PBMCs but not NK cell-depleted PBMCs treated with LEN augmented 30% of RTX-dependent cytotoxicity. Daily treatment with LEN increased NK cells by 10-folds in SCID mice, and combination of LEN and RTX decreased tumor burden and prolonged survival of MCL-bearing SCID mice. Taken together, our study demonstrates that LEN plus RTX provides a synergistically therapeutic effect on MCL cells by enhancing apoptosis and RTX-dependent NK cell mediated cytotoxicity and may be an optimal combination in the clinical trial of relapsed or refractory MCL. PMID- 19565650 TI - Hypothesis: Bifunctional mitochondrial proteins have centrosomal functions. AB - Mitochondria are dynamic organelles that are involved in a number of diverse processes. Most often the mitochondrion is associated with energy generation, but other important processes occur in this organelle such as fatty acid synthesis and amino acid metabolism. Although mitochondria encode less than 40 genes, all of the other approximately 1,000 genes required for their function are nuclear encoded. The protein products from these nuclear encoded genes are subsequently translocated to the mitochondria and utilized in the variety of processes within the organelle. Is it possible then that any of these nuclear encoded proteins could be translocated to other areas of the cell to participate in functions not normally attributed to the mitochondria? There is growing evidence that mitochondrial proteins not only localize to sites outside of the organelle, but also have functionality at these other locales. We suggest that a subset of nuclear encoded mitochondrial proteins are bifunctional and are involved in processes outside of the mitochondria. In this perspective, we will discuss published data demonstrating mitochondrial proteins that influence progression of the cell cycle, alter chromosome morphology, localize to centrosomes, and affect centrosome number in the cell. Overall, the mitochondrion is an interesting organelle that participates in a variety of vital functions. We suggest that these essential functions are not solely due to the ability of the mitochondria to produce high amounts of energy. Throughout this discussion we attempt to demonstrate that a cell cannot live without mitochondria and study this issue from a nonenergetic perspective. PMID- 19565651 TI - Anticonvulsant and neuroprotective effects of the novel calcium antagonist NP04634 on kainic acid-induced seizures in rats. AB - Kainic acid (KA)-induced status epilepticus (SE) is a well-characterized model of excitotoxic neuronal injury. Excitotoxicity results from activation of specific glutamate receptors, with resultant elevation of intracellular Ca(2+). The CA1 and CA3 subregions of the hippocampus are especially vulnerable to KA, and this pattern of neuronal injury resembles that occurring in patients with temporal lobe epilepsy. Calcium plays an essential role in excitotoxicity, and accordingly calcium channel inhibitors have been shown to have protective effects in various experimental models of epilepsy and brain injury. Moreover, they also potentiate the antiseizure efficacy of conventional antiepileptic drugs. This study was undertaken to determine whether NP04634, a novel compound, reported as a non-L type voltage-sensitive calcium channel (VSCC) inhibitor, could prevent the entrance in SE and the neuronal loss evoked by intraperitoneal injection of KA. Our results show that intragastrical administration of NP04634 reduced the percentage of rats that entered SE after KA injection, increased the latency of SE entry, and significantly reduced the mortality of rats that entered SE. Also, NP04634 prevented the loss of hippocampal CA1 and CA3 pyramidal neurons and reduced the gliosis induced by KA. These results point to a potential anticonvulsant and neuroprotective role for NP04634. PMID- 19565652 TI - Environmental enrichment stimulates progenitor cell proliferation in the amygdala. AB - Enriched environments enhance hippocampal neurogenesis, synaptic efficacy, and learning and memory functions. Recent studies have demonstrated that enriched environments can restore learning behavior and long-term memory after significant brain atrophy and neural loss. Emotional and anxiety-related behaviors were also improved by enriched stimuli, but the effect of enriched environments on the amygdala, one of the major emotion-related structures in the central nervous system, remains largely unknown. In this study, we have focused on the effects of an enriched environment on cell proliferation and differentiation in the murine amygdala. The enriched environment increased bromodeoxyuridine (BrdU)-positive (newborn) cell numbers in the amygdala, almost all of which, immediately after a 1-week period of enrichment, expressed the oligodendrocyte progenitor marker Olig2. Furthermore, enriched stimuli significantly suppressed cell death in the amygdala. Some of the BrdU-positive cells in mice exposed to the enriched environment, but none in animals housed in the standard environment, later differentiated into astrocytes. Our findings, taken together with previous behavioral studies, suggest that progenitor proliferation and differentiation in the amygdala may contribute to the beneficial aspects of environmental enrichment such as anxiolytic effects. PMID- 19565653 TI - Differential clustering of Caspr by oligodendrocytes and Schwann cells. AB - Formation of the paranodal axoglial junction (PNJ) requires the presence of three cell adhesion molecules: the 155-kDa isoform of neurofascin (NF155) on the glial membrane and a complex of Caspr and contactin found on the axolemma. Here we report that the clustering of Caspr along myelinated axons during development differs fundamentally between the central (CNS) and peripheral (PNS) nervous systems. In cultures of Schwann cells (SC) and dorsal root ganglion (DRG) neurons, membrane accumulation of Caspr was detected only after myelination. In contrast, in oligodendrocytes (OL)/DRG neurons cocultures, Caspr was clustered upon initial glial cell contact already before myelination had begun. Premyelination clustering of Caspr was detected in cultures of oligodendrocytes and retinal ganglion cells, motor neurons, and DRG neurons as well as in mixed cell cultures of rat forebrain and spinal cords. Cocultures of oligodendrocyte precursor cells isolated from contactin- or neurofascin-deficient mice with wild type DRG neurons showed that clustering of Caspr at initial contact sites between OL processes and the axon requires glial expression of NF155 but not of contactin. These results demonstrate that the expression of membrane proteins along the axolemma is determined by the type of the contacting glial cells and is not an intrinsic characteristic of the axon. PMID- 19565654 TI - Spinal anterior horn has the capacity to self-regenerate in amyotrophic lateral sclerosis model mice. AB - The exact host environment necessary for neural regeneration in amyotrophic lateral sclerosis (ALS) has not yet been fully elucidated. We first focused on the extracellular matrix proteins in ALS model mice during development of the disease and then attempted to examine whether regeneration occurs in the ALS spinal cord under regenerative conditions. A progressive increase in gamma1 laminin (a promoter of regeneration) and a progressive decrease in semaphorin3A (Sema3A; an inhibitor of regeneration) were observed, mainly in the neuropil of the spinal anterior horn from 15 to 18 weeks, when astrocytes began to express both gamma1 laminin and Sema3A. On the other hand, a progressive increase in growth-associated protein 43 (GAP43; synaptic regeneration site) and a progressive decrease in synaptotagmin1 (actual synaptic bouton) were observed in the same areas of the spinal anterior horn from 15 to 18 weeks. Thus, the present data suggest that, although the spinal anterior horn in ALS models loses motor neurons, it initially possesses the capacity to self-regenerate but displays a progressive loss of ability to regenerate new effective synapses. PMID- 19565655 TI - Sodium channelopathy induced by mild axonal trauma worsens outcome after a repeat injury. AB - There is great concern that one mild traumatic brain injury (mTBI) predisposes individuals to an exacerbated response with a subsequent mTBI. Although no mechanism has been identified, mounting evidence suggests traumatic axonal injury (TAI) plays a role in this process. By using a cell culture system, a threshold of mild TAI was found where dynamic stretch of cortical axons at strains lower than 5% induced no overt pathological changes. However, the axons were found to display an increased expression of sodium channels (NaChs) by 24 hr. After a second, identical mild injury, pathologic increases in [Ca(2+)](i) were observed, leading to axon degeneration. The central role of NaChs in this response was demonstrated by blocking NaChs with tetrodotoxin prior to the second injury, which completely abolished postinjury increases in [Ca(2+)](i). These data suggest that mild TAI induces a form of sodium channelopathy on axons that greatly exaggerates the pathophysiologic response to subsequent mild injuries. PMID- 19565656 TI - Down-regulation of Bcl-2 enhances autophagy activation and cell death induced by mitochondrial dysfunction in rat striatum. AB - In vivo administration of the mitochondrial inhibitor 3-nitropropionic acid (3 NP) produces striatal pathology mimicking Huntington's disease (HD). However, the mechanisms of cell death induced by metabolic impairment are not fully understood. Previous studies showed that 3-NP triggered p53-depedent autophagy activation and cell death. The present study investigated the contribution of the Bcl-2 signaling pathway to autophagy activation and cell death induced by 3-NP. Rat striatum was intoxicated with 3-NP by stereotaxic injection. 3-NP up regulated the expression of the autophagic protein beclin 1 but down-regulated the expression of the antiapoptotic protein Bcl-2. Pretreatment with the autophagy inhibitor 3-methyladenine (3-MA) significantly inhibited the 3-NP induced alterations in beclin 1 and Bcl-2 protein levels. Similarly, the 3-NP induced decline in Bcl-2 was also prevented by the lysosomal inhibitor E64, indicating degradation of Bcl-2 by lysosomes. In agreement with the time course of 3-NP-induced cell death, an increase in the release of cytochrome c from mitochondria was observed. 3-MA also attenuated the 3-NP-induced release of cytochrome c. On the other hand, 3-NP-induced elevations in proapoptotic protein Bax and autophagic protein beclin 1 and LC3-II were significantly enhanced by the Bcl-2-specific inhibitor HA14-1. Furthermore, HA14-1 increased the release of cytochrome c and 3-NP-induced striatal damage. These results suggest that induction of autophagy leads to degradation of Bcl-2. Meanwhile, down-regulation of Bcl-2 amplifies autophagy activation and apoptotic signaling. Bcl-2 thus plays important roles in mitochondria dysfunction-induced apoptotic death of stritatal neurons by modulating both autophagic and apoptotic processes. PMID- 19565657 TI - Proteasome inhibition modulates kinase activation in neural cells: relevance to ubiquitination, ribosomes, and survival. AB - In this study we examined whether established signal transduction cascades, p44/42 mitogen-activated protein kinase (ERK1/2) and Jun N-terminal kinases (JNK) pathways, are altered in N2a neural cells in response to proteasome inhibition. Additionally, we sought to elucidate the relative contribution of these signal transduction pathways to the multiple downstream effects of proteasome inhibition. Our data indicate that ERK1/2 and JNK are activated in response to proteasome inhibition. Washout of proteasome inhibitor (MG132) results in an enhancement of ERK1/2 activation and amelioration of JNK activation. Treatment with an established MAPK inhibitor resulted in an increase in proteasome inhibitor toxicity, and incubation with JNK inhibitor was observed to attenuate proteasome inhibitor toxicity significantly. Subsequent studies demonstrated that inhibition of ERK1/2 and JNK activity does not alter the gross increase in ubiquitinated protein following proteasome inhibitor administration. Similarly, ERK1/2 and JNK activity do not appear to play a role in the disruption of polysomes following proteasome inhibitor administration in neural cells. Together these data indicate that ERK1/2 and JNK activation may play differential roles in modulating neurochemical disturbances and neurotoxicity induced by proteasome inhibition. PMID- 19565658 TI - Loss of astrocytic glutamate transporters in Wernicke encephalopathy. AB - Wernicke encephalopathy (WE), a neurological disorder caused by thiamine deficiency (TD), is characterized by structural damage in brain regions that include the thalamus and cerebral cortex. The basis for these lesions is unclear, but may involve a disturbance of glutamatergic neurotransmission. We have therefore investigated levels of the astrocytic glutamate transporters EAAT1 and EAAT2 in order to evaluate their role in the pathophysiology of this disorder. Histological assessment of the frontal cortex revealed a significant loss of neurons in neuropathologically confirmed cases of WE compared with age-matched controls, concomitant with decreases in alpha-internexin and synaptophysin protein content of 67 and 52% by immunoblotting. EAAT2 levels were diminished by 71% in WE, with levels of EAAT1 also reduced by 62%. Loss of both transporter sites was confirmed by immunohistochemical methods. Development of TD in rats caused a profound loss of EAAT1 and EAAT2 in the thalamus accompanied by decreases in other astrocyte-specific proteins. Treatment of TD rats with N acetylcysteine prevented the downregulation of EAAT2 in the medial thalamus, and ameliorated the loss of several other astrocyte proteins, concomitant with increased neuronal survival. Our results suggest that (1) loss of EAAT1 and EAAT2 glutamate transporters is associated with structural damage to the frontal cortex in patients with WE, (2) oxidative stress plays an important role in this process, and (3) TD has a profound effect on the functional integrity of astrocytes. Based on these findings, we recommend that early treatment using a combination of thiamine AND antioxidant approaches should be an important consideration in cases of WE. PMID- 19565659 TI - Sciatic nerve injury induces apoptosis of dorsal root ganglion satellite glial cells and selectively modifies neurosteroidogenesis in sensory neurons. AB - Neurosteroids are synthesized either by glial cells, by neurons, or within the context of neuron-glia cross-talk. Various studies suggested neurosteroid involvement in the control of neurodegeneration but there is no evidence showing that the natural protection of nerve cells against apoptosis directly depends on their own capacity to produce neuroprotective neurosteroids. Here, we investigated the interactions between neurosteroidogenesis and apoptosis occurring in sensory structures of rats subjected to neuropathic pain generated by sciatic nerve chronic constriction injury (CCI). Using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), we observed no apoptotic cells in the spinal cord up to 30 days after CCI although pain symptoms such as mechano-allodynia, thermal and mechanical hyperalgesia were evidenced with the Hargreaves's behavioral and von Frey filament tests. In contrast, double-labeling experiments combining TUNEL and immunostaining with antibodies against glutamine synthetase or neuronal nuclei protein revealed apoptosis occurrence in satellite glial cells (SGC) (not in neurons) of CCI rat ipsilateral dorsal root ganglia (DRG) at day 30 after injury. Pulse-chase experiments coupled with high performance liquid chromatography and flow scintillation detection showed that, among numerous biosynthetic pathways converting [(3)H]pregnenolone into various [(3)H]neurosteroids, only [(3)H]estradiol formation was selectively modified and upregulated in DRG of CCI rats. Consistently, immunohistochemical investigations localized aromatase (estradiol-synthesizing enzyme) in DRG neurons but not in SGC. Pharmacological inhibition of aromatase caused apoptosis of CCI rat DRG neurons. Altogether, our results suggest that endogenously produced neurosteroids such as estradiol may be pivotal for the protection of DRG sensory neurons against sciatic nerve CCI induced apoptosis. PMID- 19565660 TI - Anandamide enhances IL-10 production in activated microglia by targeting CB(2) receptors: roles of ERK1/2, JNK, and NF-kappaB. AB - The endocannabinoid system exhibits anti-inflammatory properties by regulating cytokine production. Anandamide (AEA) down-regulates proinflammatory cytokines in a viral model of multiple sclerosis (MS). However, little is known about the mechanisms by which AEA exerts these effects. Microglial cells are the main source of cytokines within the brain and the first barrier of defense against pathogens by acting as antigen presenting cells. IL-10 is a key physiological negative regulator of microglial activation. In this study we show that AEA enhances LPS/IFNgamma-induced IL-10 production in microglia by targeting CB(2) receptors through the activation of ERK1/2 and JNK MAPKs. AEA also inhibits NF kappaB activation by interfering with the phosphorylation of IkappaBalpha, which may result in an increase of IL-10 production. Moreover, endogenously produced IL 10 negatively regulates IL-12 and IL-23 cytokines, which in its turn modify the pattern of expression of transcription factors involved in Th commitment of splenocytes. This suggests that by altering the cytokine network, AEA could indirectly modify the type of immune responses within the central nervous system (CNS). Accordingly, pharmacological modulation of AEA uptake and degradation might be a useful tool for treating neuroinflammatory diseases. PMID- 19565661 TI - Biocompatible nanoparticles trigger rapid bacteria clustering. AB - This study reveals an exciting phenomenon of stimulated bacteria clustering. Rapid aggregation and microbial arrest are shown to occur in Escherichia coli solutions of neutral pH when chitosan nanoparticles with positive zeta potential are added. Because chitosan nanoparticles can easily be dispersed in aqueous buffers, the rapid clustering phenomenon requires only minuscule nanoparticle concentrations and will be critical in developing new methods for extricating bacterial pathogens. This work establishes the dominant role of electrostatic attraction in bacteria-nanoparticle interactions by varying the nanoparticle zeta potential from highly positive to strongly negative values, and by exploring concentration effects. For strongly negative nanoparticles, no clusters form, while aggregates are small and loose at intermediate conditions. In addition, optical density measurements indicate that over 90% of the suspended bacteria flocculate within seconds of being mixed with chitosan nanoparticles of a highly positive surface charge. Finally, the nanoparticles are significantly more efficient as a clustering agent compared to an equal mass of molecular chitosan in solution, as the bacteria-nanoparticle clusters formed are substantially larger. The bacteria-nanoparticle aggregation effect demonstrated here promises a rapid separation method for aiding pathogen detection and for flocculation of bacteria in fermentation processes. PMID- 19565662 TI - Perfluorocarbon facilitated O(2) transport in a hepatic hollow fiber bioreactor. AB - A mathematical model describing O(2) transport in a hepatic hollow fiber (HF) bioreactor supplemented with perfluorocarbons (PFCs) in the circulating cell culture media was developed to explore the potential of PFCs in properly oxygenating a bioartificial liver assist device (BLAD). The 2-dimensional model is based on the geometry of a commercial HF bioreactor operated under steady state conditions. The O(2) transport model considers fluid motion of a homogeneous mixture of cell culture media and PFCs, and mass transport of dissolved O(2) in a single HF. Each HF consists of three distinct regions: (1) the lumen (conducts the homogeneous mixture of cell culture media and PFCs), (2) the membrane (physically separates the lumen from the extracapillary space (ECS), and (3) the ECS (hepatic cells reside in this compartment). In a single HF, dissolved O(2) is predominantly transported in the lumen via convection in the axial direction and via diffusion in the radial direction through the membrane and ECS. The resulting transport equations are solved using the finite element method. The calculated O(2) transfer flux showed that supplementation of the cell culture media with PFCs can significantly enhance O(2) transport to the ECS of the HF when compared with a control with no PFC supplementation. Moreover, the O(2) distribution and subsequent analysis of ECS zonation demonstrate that limited in vivo-like O(2) gradients can be recapitulated with proper selection of the operational settings of the HF bioreactor. Taken together, this model can also be used to optimize the operating conditions for future BLAD development that aim to fully recapitulate the liver's varied functions. PMID- 19565663 TI - p75 neurotrophin receptor is implicated in the ability of neurotrophin-3 to negatively modulate activated ERK1/2 signaling in TrkA-expressing adult sensory neurons. AB - Neurotrophin-3 (NT-3) can negatively modulate trkA and associated phenotype in intact sensory neurons, while positively regulating trkC and associated phenotype. How NT-3 effects this response is not clear. Whether NT-3 exerts a differential influence on levels of activated ERK1/2 signaling in trkA- versus trkC-mRNA-positive subpopulations of neurons and the role that the common neurotrophin receptor, p75NTR, plays in this response was assessed by examining alterations in the levels of phospho-ERK1/2 immunofluorescence signal over nuclei of sensory neurons expressing trkA alone, trkC alone, or both trkA and trkC mRNA. NT-3 intrathecal infusion differentially modulated nuclear phospho-ERK1/2 levels detected over neurons expressing trkA alone or trkC alone. Levels were significantly decreased over nuclei of neurons expressing trkA alone and significantly increased over the nuclei of neurons expressing trkC alone. Neurons expressing both trkA and trkC or neurons expressing neither trkA nor trkC had no significant alteration in phospho-ERK1/2. Antisense oligonucleotides directed against p75NTR were infused intrathecally with or without NT-3 infusion to examine the impact of suppressing p75NTR expression on the ability of NT-3 to diminish phospho-ERK1/2 signaling in neurons expressing only trkA. NT-3 did not significantly attenuate levels of phospho-ERK1/2 when p75NTR expression was suppressed by antisense infusion, despite being able to do so when NT-3 was infused alone. In conclusion, NT-3's ability to negatively modulate ERK1/2 signaling in a p75-dependent manner in sensory neurons that express trkA to the exclusion of trkC provides a feasible mechanism by which it negatively modulates other aspects of nociceptive phenotype in these neurons. PMID- 19565665 TI - Efferent innervation to the auditory basilar papilla of scincid lizards. AB - Hair cells of the inner ear of vertebrates are innervated by afferent neurons that transmit sensory information to the brain as well as efferent neurons that receive feedback from the brainstem. The function of the efferent feedback system is poorly understood and may have changed during evolution when different tetrapod groups acquired sensitivity to airborne sound and extended their hearing ranges to higher frequencies. Lizards show a unique subdivision of their basilar papilla (homologous to the mammalian organ of Corti) into a low-frequency (<1 kHz) and a high-frequency (approximately 1-5 kHz) region. The high-frequency region was reported to have lost its efferent innervation, suggesting it was insignificant or even functionally detrimental at higher frequencies. We re examined the innervation to the basilar papilla of five species of Australian scincid lizards, by using immunohistochemistry. Anti-choline acetyltransferase (ChAT) was used as an efferent marker. Co-localization with anti-synaptic vesicle protein 2 confirmed the synaptic identity of label. Cholinergic terminals were observed along the whole length of the basilar papilla, including the regions that had previously been described as devoid of efferent innervation. However, there was a clear decrease in terminal density from apical, low-frequency to basal, high-frequency locations. Our findings suggest that efferent innervation is a general feature of the hair cells in the basilar papilla of lizards, irrespective of tonotopic location. This re-enforces the notion that efferent feedback control of hair cells is a fundamental and important property of all vertebrate hearing organs. PMID- 19565666 TI - By the way, doctor. We have screening tests for other cancers. How about pancreatic cancer? PMID- 19565664 TI - Peptidergic clock neurons in Drosophila: ion transport peptide and short neuropeptide F in subsets of dorsal and ventral lateral neurons. AB - About 150 clock neurons are clustered in different groups in the brain of Drosophila. Among these clock neurons, some pigment-dispersing factor (PDF) positive and PDF-negative lateral neurons (LNs) are principal oscillators responsible for bouts of activity in the morning and evening, respectively. The full complement of neurotransmitters in these morning and evening oscillators is not known. By using a screen for candidate neuromediators in clock neurons, we discovered ion transport peptide (ITP) and short neuropeptide F (sNPF) as novel neuropeptides in subpopulations of dorsal (LN(d)s) and ventral (s-LN(v)s) LNs. Among the six LN(d)s, ITP was found in one that coexpresses long neuropeptide F (NPF) and cryptochrome. We detected sNPF in two LN(d)s that also express cryptochrome; these cells are distinct from three LN(d)s expressing NPF. Thus, we have identified neuropeptides in five of the six LN(d)s. The three LN(d)s expressing cryptochrome, with either ITP or sNPF, are the only ones with additional projections to the accessory medulla. Among the five s-LN(v)s in the adult brain, ITP was detected in the fifth neuron that is devoid of PDF and sNPF in the four neurons that also express PDF. By using a choline acetyltransferase (Cha) Gal4, we detected Cha expression in the two sNPF producing LN(d)s and in the fifth s-LN(v). In the larval brain, two of the four PDF-producing s-LN(v)s coexpress sNPF. Our findings emphasize that the LN(d)s are heterogeneous both anatomically and with respect to content of neuropeptides, cryptochrome, and other markers and suggest diverse functions of these neurons. PMID- 19565667 TI - Two different doses of caudal neostigmine co-administered with levobupivacaine produces analgesia in children. AB - SUMMARY BACKGROUND: This study was aimed to evaluate the analgesic efficacy duration of analgesia, and side effects of two different doses of caudal neostigmine used with levobupivacaine in children. METHODS: Sixty boys, between 5 months and 5 years, undergoing genitourinary surgery were allocated randomly to one of three groups (n =20 each). Group I patients received caudal 0.25% levobupivacaine (1 ml.kg(-1)) alone. Groups II and III patients received neostigmine (2 and 4 microg.kg(-1) respectively) together with levobupivacaine used in the same does as Group I. Pain scores were assessed using Children's and Infant's Postoperative Pain Scale (CHIPPS) at 15th (t(1)) min after arrival to postanesthetic care unit, and 1st (t(2)), 2nd (t(3)), 3rd (t(4)), 4th (t(5)), 8th (t(6)), 16th (t(7)), and 24th (t(8)) hour postoperatively. Duration of analgesia, amount of additional analgesic (paracetamol), score of motor blockade and complications were recorded for 24 h postoperatively, and compared between groups. RESULTS: CHIPPS scores were higher during t(2), t(3), t(6), t(7), and t(8) periods, duration of analgesia was shorter, and total analgesic consumption was higher in Group I compare to neostigmine groups (P < 0.05). Duration of postoperative analgesia and total analgesic consumption were similar in Groups II and III (P > 0.05). Adverse effects were not different between three groups. CONCLUSIONS: Caudal neostigmine in doses of 2 and 4 microg.kg(-1) with levobupivacaine extends the duration of analgesia without increasing the incidence of adverse effects, and 2 microg.kg(-1) seems to be the optimal dose, as higher dose has no further advantages. PMID- 19565668 TI - Two functions of early language experience. AB - The unique human ability of linguistic communication, defined as the ability to produce a practically infinite number of meaningful messages using a finite number of lexical items, is determined by an array of "linguistic" genes, which are expressed in neurons forming domain-specific linguistic centers in the brain. In this review, I discuss the idea that infants' early language experience performs two complementary functions. In addition to allowing infants to assimilate the words and grammar rules of their mother language, early language experience initiates genetic programs underlying language production and comprehension. This hypothesis explains many puzzling characteristics of language acquisition, such as the existence of a critical period for acquiring the first language and the absence of a critical period for the acquisition of additional language(s), a similar timetable for language acquisition in children belonging to families of different social and cultural status, the strikingly similar timetables in the acquisition of oral and sign languages, and the surprisingly small correlation between individuals' final linguistic competence and the intensity of their training. Based on the studies of microcephalic individuals, I argue that genetic factors determine not only the number of neurons and organization of interneural connections within linguistic centers, but also the putative internal properties of neurons that are not limited to their electrophysiological and synaptic properties. PMID- 19565669 TI - Immunocytochemical detection of synaptophysin in C57BL/6 mice cochlea during aging process. AB - Aged mammals frequently exhibit a bilateral, progressive, and symmetric deafness related to the degeneration of auditory receptor. However, little is still known about aging effects on synapses in this receptor. Synaptophysin (Syp) is a 38 kDa Ca2+ binding glycoprotein widely found in presynaptic membrane and vesicles. The Syp has been found in presynaptic buttons of efferent auditory fibers, within the developing and adult auditory receptor. The detection of Syp in aged cochleae could provide relevant information about synaptic changes and receptor degeneration process observed in old animals. This paper focuses on aging linked changes related to the presence of Syp in cochleae of C57BL/6J mice (from 1 to 24 months old). Results showed that during the first months of age, no significant changes were observed in the Syp distribution under the basal pole of inner (IHCs) neither the outer (OHCs) hair cells. At six months of age, a significant decrease of Syp immunocytochemical detection appeared in fibers under the most external row of OHCs, but restricted to the cochlear basal coil. Only a very scarce reduction of Syp was noted under the IHC and the other OHC rows, also at the basal coil. From mice 9 months old on, a progressive decrease of the presence of Syp was found under IHC and all OHC rows starting at the basal coil and reaching the apical coil in the oldest mice. All these data could indicate that the cochlea aging process early affects to presynaptic membrane proteins of efferent endings fibers. This early alteration of cochleae efferent synapses could be involved in the whole degeneration of the Corti's organ. PMID- 19565670 TI - A ring enhancing lesion. PMID- 19565671 TI - Chest X-ray quiz. Gastric distension. PMID- 19565672 TI - Immunoregulatory effects of indoleamine 2, 3-dioxygenase in transplantation. AB - Indoleamine 2, 3-dioxygenase (IDO) is an intracellular hemeprotein enzyme which catalyses the essential amino acid tryptophan. Accumulating evidence has demonstrated that tryptophan depletion and its toxic metabolites expression in tissue microenvironment can suppress local allogeneic T cell proliferation and activation. Ever since the discovery that IDO was involved in the maintenance of fetal-maternal tolerance, numerous studies have confirmed that IDO is a potent regulator of immune cell function. Importantly, IDO+dendritic cells (DCs) might interact with regulatory T cells (Tregs) to form an immunomodulatory network to promote immune tolerance induction. Moreover, it has been reported that overexpression of IDO in transplanted organs can prolong allograft survival, suggesting a possible peripheral tolerogenic pathway with important implications in transplantation. However, the underlying mechanism for the beneficial effects of IDO in transplantation remains unclear. In this review, we attempt to summarize our current understandings about IDO as a mediator of immunity in transplantation and provide an overview of IDO as a new paradigm in transplantation. PMID- 19565674 TI - Retraction notice to "The immune decision toward allograft tolerance in nonhuman primates requires early inhibition of innate immunity and induction of immune regulation" [Transpl Immunol 11 (2003) 335-344]. PMID- 19565673 TI - The significance of EMAP-II after allogeneic stem cell transplantation. AB - Endothelial monocyte-activating polypeptide-II (EMAP-II) appears to play an important role in neovascularization and endothelial abnormalities. However, the role of EMAP-II in development of graft-versus-host disease (GVHD) after allogeneic SCT is poorly understood. We measured and compared the levels of EMAP II, cytokines, and soluble factors in patients undergoing allogeneic SCT. The subjects were 23 patients who underwent allogeneic SCT. Most of the cytokines/soluble factors exhibited a significant elevation after allogeneic SCT, although Angiopoietin-1 did not change. On the other hand, the levels of these factors did not change significantly in the recipients of autologous SCT. When the relationship between EMAP-II and cytokines/soluble factors was analyzed, EMAP II levels correlated positively with sIL-2R, sVCAM-1, sE-selectin, sFasL and EDMP. However, IL-6, Angiopoietin-1, Angiopoietin-2 and VEGF were not correlated with EMAP-II. Our results suggest that EMAP-II plays an important role in endothelial cell dysfunction related to GVHD after allogeneic SCT. PMID- 19565675 TI - Phylogeography of the malagasy ring-tailed mongoose, Galidia elegans, from mtDNA sequence analysis. AB - The ring-tailed mongoose (Galidia elegans) represents one of the most widely distributed mongooses in Madagascar; however, we know little about the ecology of this seemingly ubiquitous species. Currently, G. elegans is divided into three recognized subspecies--G. e. elegans, G. e. dambrensis, and G. e. occidentalis- based on differences in pelage coloration between the distinct geographic locations. We used intraspecific DNA variation to describe the phylogenetic relationships among the described subspecies. Approximately 550 base pairs of the mitochondrial DNA control region were analyzed from 19 G. elegans specimens representing all three subspecies sampled from across the species' geographic range. Sequence data from outgroup taxa were included for comparison. Examination of the recovered sequences revealed a strongly supported distinct genetic signature in the western region of the island, but remained inconclusive with respect to supporting the designation of the northern and eastern 'subspecies' for treatment as divergent intraspecific units for management. PMID- 19565677 TI - Complaints against doctors. PMID- 19565676 TI - The mitochondrial trn-cox1 locus: rapid evolution in Pompilidae and evidence of bias in cox1 initiation and termination codon usage. AB - BACKGROUND: The translation initiation codon of the mitochondrial (mt) cytochrome oxidase I gene, cox1, frequently differs from conventional AUN triplets, and the termination codon is often incomplete. METHODS: To investigate whether initiation and termination codon usage may depend on the rate of mt genome evolution, a fast evolving locus comprising cox1 and adjacent tRNA-encoding trn genes was sequenced from representatives of the major clades of spider wasps, Pompilidae (Hymenoptera), and then included in insect-wide analyses of codon usage in the context of mtDNA sequence stability. RESULTS: The rapid evolution of Pompilidae trn-cax1 locus was particularly evident in the rearrangement of the trnC and trnY genes, the presence of a trn pseudogene in Psorthaspis, and the relatively high frequency of cox1 nucleotide substitutions. Such extensive genetic diversification, underscored by variability in the arrangement of trn genes even at the subfamily level, contrasted with the conventional form of cox1 initiation and termination codons. Similar preference for codon usage was identified for other fast evolving insect lineages unrelated to Pompilidae. Instead, conventional codons were often absent in slower evolving loci. CONCLUSION: Analysis of codon usage across a spectrum of evolutionary rates indicates the existence of positive correlation between the incidence of cox1 canonical initiation and termination codons and relative rate of the trn-cox1 locus evolution. PMID- 19565678 TI - Phosphorylation puts sperm in the mood. PMID- 19565680 TI - Finite element modelling of medical devices. Proceedings of the Summer Workshop of the European Society of Biomechanics, August 2007, Dublin, Ireland. PMID- 19565679 TI - An unusual parahippocampal lesion presenting with seizures. PMID- 19565681 TI - Motherhood: the early days. PMID- 19565682 TI - Swine flu. PMID- 19565684 TI - In conversation with John Crocker. Interviewed by Kevin Smith. PMID- 19565683 TI - A power primer. AB - One possible reason for the continued neglect of statistical power analysis in research in the behavioral sciences is the inaccessibility of or difficulty with the standard material. A convenient, although not comprehensive, presentation of required sample sizes is provided here. Effect-size indexes and conventional values for these are given for operationally defined small, medium, and large effects. The sample sizes necessary for .80 power to detect effects at these levels are tabled for eight standard statistical tests: (a) the difference between independent means, (b) the significance of a product-moment correlation, (c) the difference between independent rs, (d) the sign test, (e) the difference between independent proportions, (f) chi-square tests for goodness of fit and contingency tables, (g) one-way analysis of variance, and (h) the significance of a multiple or multiple partial correlation. PMID- 19565685 TI - Early parental adjustment and bereavement after childhood cancer death. AB - This study comprehensively explored parental bereavement and adjustment at 6 months post-loss due to childhood cancer. Interviews were conducted with 18 mothers and 13 fathers. Interviews were transcribed verbatim and analyzed based on qualitative methodology. A model describing early parental bereavement and adaptation emerged with 3 domains: (1) Perception of the Child, describing bereavement and adjustment prior to and after the loss; (2) Perception of Others, including relationships with partners, surviving children, and their social network; and (3) Perception of the World, exploring parents' perceived meanings of the experience in the context of their worldview. Domains are illustrated by quotes. Profiles of parental bereavement emerged. PMID- 19565686 TI - Portraits of caregivers of end-stage dementia patients receiving hospice care. AB - The purpose of this study was to investigate how caregivers respond to the end stages of dementia with the assistance from hospice. Data were collected from 27 family caregivers over the course of 10 months, with each caregiver being interviewed up to 4 times during the time that the patient received hospice care. Chart review data were also collected. Four distinct caregiver portraits emerged: (a) disengaged; (b) questioning; (c) all-consumed; and (d) reconciled. Caregivers in each portrait differed in how they responded to the impending death of the care recipient, the disease progression, and hospice care. Recognizing the differences in the ways that caregivers respond to the final stages of the disease will assist hospice and other providers in best meeting the needs of the caregivers. PMID- 19565687 TI - Making the transition to hospice: exploring hospice professionals' perspectives. AB - Hospice care is available for 6 months before death but the length of use varies widely, suggesting that there are different perspectives on the appropriate timing for this transition. This qualitative study explored hospice professionals' views on the appropriate timing for and communication about hospice. Ethnography of team meetings informed the development of questions for focus groups and written follow-up surveys with 53 hospice professionals. Results suggest timely admissions are situation-specific and involve a terminal prognosis, terminal decline, and a mutual understanding. Important aspects of communication among families, physicians, and hospice teams are described. PMID- 19565688 TI - Dress for the occasion. PMID- 19565689 TI - Sustaining and spreading reduced door-to-balloon times for ST-segment elevation myocardial infarction patients. AB - BACKGROUND: Prompt primary percutaneous coronary intervention (PCI) for patients with ST-segment elevation myocardial infarction (STEMI) significantly reduces mortality and morbidity. In 2004 the American College of Cardiology (ACC) and American Heart Association (AHA) set a goal to reduce door-to-balloon (D2B) time to < 90 minutes in 75% of STEMI cases. IMPLEMENTING THE STEMI INITIATIVE: In 2004, the STEMI/D2B leadership team broke down D2B time into four segments: door to data, data to diagnosis, diagnosis to decision, and decision to device. Each segment was examined for inefficiencies, duplication, and nonstandardization. In 2005, after the internal D2B processes and results showed improvement, the STEMI/D2B leadership team extended the project to prehospital emergency medical services. In 2006, UMass Memorial began to roll out a regional system for STEMI care to the 12 community hospitals in its service area without on-site PCI capabilities. RESULTS: In 2007, the STEMI program's first full year, D2B times averaged < or = 90 minutes in 94% of the 87 STEMI cases; 62% had a D2B of < or = 60 minutes. In 2008, 96% of the D2B times averaged < or = 90 minutes. Mortality rates following PCI for STEMI were 62% and 57% less than predicted in 2006 and 2007, respectively. In 2008 the D2B time for direct-admit STEMI patients averaged < 50 minutes. From December 2007 through April 2009 UMass Memorial achieved the new ACC/AHA metric of prehospital EKG to balloon in < or = 90 minutes for 64 (90%) of the 71 patients for whom a prehospital electrocardiogram was obtained. DISCUSSION: The D2B time process is being applied to other clinical venues; a vascular surgery project is underway to reduce "door-to-incision time" for patients with ruptured abdominal aortic aneurysms. PMID- 19565691 TI - Consumers' use of internet-based nursing home report cards. AB - BACKGROUND: Few studies have addressed the extent to which consumers use nursing home report cards and how they use this information. These questions, which are significant because the nursing home market presents several challenges to the effectiveness of report cards, were pursued in a study. METHODS: Data used in this investigation came from three mail surveys conducted in 2006 of family members of nursing home residents (N = 4,754), family members of assisted-living residents (N = 496), and elders living in high-rise buildings (N = 1,252). RESULTS: Some 31% of family members of nursing home residents, 53% of family members of assisted-living residents, and 23% of elders living in high-rise buildings used the Internet in looking for a nursing home. In general, these different types of consumers look for different types of nursing home information. DISCUSSION: According to the results, most consumers who used the Internet to look for a nursing home also used a report card. In addition, between 6% and 17% of consumers specifically identified using Nursing Home Compare when looking for a nursing home. The highest use of report cards was reported by family members of assisted-living residents, followed by family members of nursing home residents. Many consumers primarily used the report cards simply to find the location of nursing homes-a useful first step when examining the facility choices available. Yet relatively few consumers primarily used report cards for the arguably more important step of comparing quality information. Future research should investigate why the quality information is not used to a greater extent and what hinders consumers from using this information. PMID- 19565690 TI - Parent-driven technology for decision support in pediatric emergency care. AB - BACKGROUND: A quasi-experimental intervention study composed of control and intervention periods was conducted to determine if a parent-driven health information technology influenced completeness of documentation and adherence to evidence-based emergency care for children. METHODS: Structured chart abstraction was used to assess documentation and correctness of clinical actions at test ordering, medication prescribed for disease, and medication ordered for pain in a tertiary care pediatric emergency department and a suburban general emergency department. During the intervention periods, parents of children who presented with complaints related to otitis media, urinary tract infection, head trauma, or asthma entered data into a health information technology (ParentLink), which produced treatment plans in the context of evidence-based guidelines. RESULTS: Of 1,410 subjects analyzed, 1,072 (76%) were assigned to one of four disease categories: urinary tract infection (22%), otitis media (20%), asthma (11%) and head trauma (47%). During ParentLink use, documentation of pain significantly improved (28% incomplete [control] versus 15% [intervention], p = .003). Incorrect actions for pain treatment decreased, but not significantly (33% [control] versus 24% [intervention], p = .13). ParentLink did not influence actions for test ordering or prescribing for disease. DISCUSSION: Parent-driven health information technology intended to translate parents' knowledge into clinical practice and to support evidence-based care suggested a trend toward modest impact on pain management but did not demonstrate broad effects across diseases or care processes. The emergence and proliferation of personally controlled health records (PCHRs) presents opportunities for patients and parents to control their medical profiles. Although ParentLink is not a comprehensive PCHR, it represents a step in incorporating parent-derived information into medical decision making. PMID- 19565692 TI - Awareness and reporting of adverse drug reactions among health care professionals in Sudan. AB - BACKGROUND: Providers are often unaware of adverse drug reactions (ADRs) or may even lack basic knowledge about them. Underreporting has been attributed to time constraints, misconceptions about spontaneous reporting and bureaucratic reporting procedures, lack of information on how to report and a lack of availability of report forms, and physicians' attitudes to ADRs. This study was undertaken to determine baseline data for health care leaders' and policymakers' knowledge, attitudes, and policies related to ADRs at eight hospitals in Wad Madani, Sudan. METHODS: A random sample of participants completed the survey, which consisted of 35 closed questions and/or open-ended statements. RESULTS: Five hundred (83.3%) of the initial 600 surveys were returned, of which 475 (95%) were completed. Of the respondents, 175 (36.8%) were physicians, 100 (21.1%) were pharmacists, and 200 (42.1%) were nursing staff. The results indicated lack of polices for ADRs in most of the surveyed facilities. More than two thirds of the participants stated that they were not performing any ADR monitoring. The main reasons for not reporting ADRs were lack of knowledge on how to report (27.0%) and lack of awareness about the existence of national or international reporting systems (26.5%). Almost half (46%) of the participants reported the lack of any educational efforts for ADR prevention. DISCUSSION: Low awareness among health care professionals toward ADRs may reflect lack of basic knowledge and lack of vigilance. The study has helped promote health care professionals' ADR awareness and vigilance at the surveyed health care facilities. Education and training regarding ADRs of health professionals are warranted. Steps have been taken to develop ADR monitoring programs in collaboration with other stakeholders. PMID- 19565693 TI - Developing and evaluating a trigger response system. AB - Based on its experience in implementing a rapid response system, a Level III trauma medical center recommends that other organizations (1) involve key stakeholders in the development process, (2) develop an awareness campaign, (3) hardwire the trigger response process, (4) develop quality success measures and metrics, and (5) implement a pilot and make data-driven changes accordingly. PMID- 19565694 TI - Water molds of the genera Saprolegnia and Leptolegnia are pathogenic to the North American frogs Rana catesbeiana and Pseudacris crucifer, respectively. AB - Water molds are commonly associated with amphibian mortality. Since water molds often act as saprophytes, it is important to test their effects on amphibians to determine whether they can also act as pathogens. In controlled experiments, the eggs of 2 amphibian species, the American bullfrog Rana catesbeiana and the spring peeper Pseudacris crucifer, suffered higher mortality when they were exposed to zoospores of water molds of the genera Saprolegnia and Leptolegnia, respectively. Water molds are important pathogens in many amphibian systems, yet their ecological impact on amphibians remains mostly unknown. PMID- 19565695 TI - Phylogenetic relationships in the family Alloherpesviridae. AB - Phylogenetic relationships among herpesviruses (HVs) of mammals, birds, and reptiles have been studied extensively, whereas those among other HVs are relatively unexplored. We have reconstructed the phylogenetic relationships among 13 fish and amphibian HVs using maximum likelihood and Bayesian analyses of amino acid sequences predicted from parts of the DNA polymerase and terminase genes. The relationships among 6 of these viruses were confirmed using the partial DNA polymerase data plus the complete sequences of the terminase, helicase, and triplex protein genes; the position of these viruses among all other sequenced HVs was also investigated using the complete terminase gene. The results established the monophyly of the fish and amphibian HVs (Alloherpesviridae) separate from the HVs of mammals, birds, and reptiles (Herpesviridae) and the single recognized HV of bivalve mollusks (Malacoherpesviridae) in the order Herpesvirales. Two major clades in the family Alloherpesviridae were recognized: one consisting of viruses from cyprinid and anguillid hosts and the other of viruses from ictalurid, salmonid, acipenserid, and ranid hosts. A comparison of virus and host phylogenies suggested that closely related HVs in this family may have coevolved with their hosts, whereas significant codiversification was not apparent for the more distantly related viruses. PMID- 19565696 TI - Detection of Laem-Singh virus in cultured Penaeus monodon shrimp from several sites in the Indo-Pacific region. AB - Laem-Singh virus (LSNV) is a positive-sense single-stranded RNA (ssRNA) virus that was recently identified in Penaeus monodon shrimp in Thailand displaying signs of slow growth syndrome. A total of 326 shrimp collected between 1998 and 2007 from countries in the Indo-Pacific region were tested by RT-PCR for evidence of LSNV infection. The samples comprised batches of whole postlarvae, and lymphoid organ, gill, muscle or pleopod tissue of juvenile, subadult and adult shrimp. LSNV was not detected in 96 P. monodon, P. japonicus or P. merguiensis from Australia or 16 P. monodon from Fiji, Philippines, Sri Lanka and Mozambique. There was no evidence of LSNV infection in 73 healthy juvenile P. vannamei collected during 2006 from ponds at 9 locations in Thailand. However, LNSV was detected in each of 6 healthy P. monodon tested from Malaysia and Indonesia, 2 of 6 healthy P. monodon tested from Vietnam and 39 of 40 P. monodon collected from slow-growth ponds in Thailand. A survey of 81 P. monodon collected in 2007 from Andhra Pradesh, India, indicated 56.8% prevalence of LSNV infection but no clear association with disease or slow growth. Phylogenetic analysis of PCR amplicons obtained from samples from India, Vietnam, Malaysia and Thailand indicated that nucleotide sequence variation was very low (>98% identity) and there was no clustering of viruses according to site of isolation or the health status of the shrimp. The data suggests that LSNV exists as a single genetic lineage and occurs commonly in healthy P. monodon in parts of Asia. PMID- 19565697 TI - Influence of temperature shifts on the onset and development of red sea bream iridoviral disease in rock bream Oplegnathus fasciatus. AB - The effects of various water temperature treatments on the development of red sea bream iridovirus disease (RSIVD) in rock bream Oplegnathus fasciatus challenged with iridovirus Sachun (IVS-1) were determined by measuring the mortality and the viral concentration in the spleen of infected fish. Experimental infections of rock bream with IVS-1 at water temperatures of 18, 21, and 25 degrees C resulted in a cumulative mortality of 100%, but infections at 13 degrees C resulted in 0% mortality, even after 45 d. The disease progressed more rapidly at higher water temperatures; at 25, 21, and 18 degrees C, the mean numbers of days until death were 17, 20, and 30 d, respectively. When the water temperature for fish infected with iridovirus by intramuscular injection was shifted from 13 to 25 degrees C, the cumulative mortality reached 100%, with rapid onset of the disease, independent of the time at which the temperature was shifted, i.e. 7, 14, or 30 d after injection at 13 degrees C. Real-time PCR data revealed that the viral genome copy number in the spleen of rock bream maintained at 13 degrees C increased with time, suggesting the occurrence of viral replication even at 13 degrees C. In the reverse experiment, when the water temperature for fish that were infected at a higher temperature was shifted to 13 degrees C, 3 or 7 d after injection at 25 degrees C, the fish showed 100% cumulative mortality, although the mean number of days until death was higher than that observed for fish maintained at a constant temperature of 25 degrees C. The viral DNA concentration in the spleen of rock bream that had been shifted down to 13 degrees C, 3 or 7 d after injection at 25 degrees C, was not suppressed, but increased and eventually reached levels sufficient to induce mortality at 13 degrees C. However, the level of viral genome copy numbers in the spleen of dead fish at 25 degrees C, regardless of whether those fish were held at a constant temperature of 25 degrees C or shifted up from 13 degrees C, appeared to be greater than the level found in the dead fish shifted down to 13 degrees C after inoculation at 25 degrees C. PMID- 19565698 TI - Identification of Vibrio harveyi isolated from diseased cultured wedge sole Dicologoglossa cuneata. AB - We report the first isolation of Vibrio harveyi from wedge sole Dicologoglossa cuneata. The pathogen was recovered from ulcers and internal organs of ailing cultured fish, from 7 different outbreaks between 2004 and 2006. The 15 isolates found were phenotypically characterized using biochemical tests and BIOLOG GN plates, which revealed high phenotypic diversity. Diagnosis was confirmed with PCR using V harveyi specific primers and partial 16S and 23S rRNA gene sequencing. A virulence evaluation of the isolates was also performed using fry and juvenile wedge sole. Significant mortalities were recorded by intraperitoneal injection; however, no mortalities were recorded by bath immersion. PMID- 19565699 TI - Route of entry and tissue distribution of Yersinia ruckeri in experimentally infected rainbow trout Oncorhynchus mykiss. AB - Yersinia ruckeri is the causative agent of enteric redmouth disease, which leads to significant losses in salmonid aquaculture worldwide. Despite the significance of the disease, little information is available on the pathogenesis. In this study, the portal of entry was investigated using a contact-exposure infection method in rainbow trout Oncorhynchus mykiss with 4 different Y. ruckeri strains. Bacteriological and histological examination revealed the presence of high numbers of bacteria in the gills immediately after infection resulting in a rapid spread of Y. ruckeri in the internal organs. However, only a virulent strain was able to survive and multiply in the host, causing septicaemia and death several days after infection. These findings indicate that gills may be an important site of entry and that Y. ruckeri virulence is related to immune evasion. PMID- 19565700 TI - Distribution, prevalence, and intensity of the swim bladder parasite Anguillicola crassus in New England and eastern Canada. AB - In the summer of 2005, yellow phase American eels Anguilla rostrata were examined for the swim bladder parasite Anguillicola crassus from 26 locations in New England, USA, ranging from the Pawcatuck River, Rhode Island, to the East Machias River, Maine. An additional 12 sites were sampled within Canada during the summers of 2006 and 2007: 7 sites in southern Nova Scotia and 5 sites within the St. Lawrence River system. In 2007, eels were also obtained from New Brunswick, northern Nova Scotia, Prince Edward Island, and Newfoundland through the commercial eel fishery. All locations in Rhode Island (n = 3) and Massachusetts (n = 10) and 7 in Maine (n = 13) had infected eels, with parasite prevalence ranging from 7 to 76%. No eels sampled from southern Nova Scotia or the St. Lawrence River system were infected with the parasite. New Brunswick and northern Nova Scotia had infected eels ranging from 3 to 30% parasite prevalence, with Cape Breton, Nova Scotia, being the furthest north the parasite has been reported in American eels. There was no significant relationship between parasite prevalence and latitude. The present study supports the hypothesis that the parasite is capable of expanding its range further into the Maritimes and could potentially reach the St. Lawrence River system. PMID- 19565701 TI - Microsporidiosis in the Caribbean spiny lobster Panulirus argus from southeast Florida, USA. AB - Two specimens of the Caribbean spiny lobster Panulirus argus captured by lobster fishers offshore of southeast Florida, USA, between late 2007 and early 2008 had completely white abdominal muscle tissue with a 'cooked' appearance. Wet-mount examination of the skeletal muscle tissue revealed masses of microsporidian spores. Histopathology of longitudinally sectioned skeletal muscle showed that the microsporidian spores displaced much of the muscle mass, but were interspersed with small empty vacuoles (approximately 5 microm in diameter) found adjacent to necrotic skeletal muscle. Skeletal muscle showed both liquefactive and coagulative necrosis. Transmission electron microscopy of the microsporidian spores revealed characteristics--including microvilli extending from the surface of the exospore, a unikaryotic spore (width 1.0 +/- 0.13 microm, range 0.8 to 1.4 microm; length 1.4 +/- 0.11 microm, range 1.2 to 1.6 microm; mean +/- SD, N = 16), and an isofilar polar filament-consistent with the genus Ameson, which is known to infect other palinurid lobsters. Microsporidiosis in Caribbean spiny lobsters has rarely been reported within the lobster's range, with only one brief report coming from the Florida Keys in 1976. Potential risks to the lobster fishery are unknown but warrant further study. PMID- 19565702 TI - Use of oxytetracycline for the treatment of tremor disease in the Chinese mitten crab Eriocheir sinensis. AB - The causative agent of tremor disease (TD) in the Chinese mitten crab Eriocheir sinensis has been shown to be a member of the genus Spiroplasma. In the present study, a susceptibility test indicated that oxytetracycline (OTC) has both a high degree of efficacy in the inhibition of Spiroplasma and a broad range of safe concentrations. Treatment experiments showed that the best concentration of OTC for use against TD was 40 mg OTC kg(-1) crab weight. Acute toxicity experiments demonstrated that the 24 and 48 h median lethal dosages (LD50) of OTC for this species of crab were 366 and 340 mg OTC kg(-1) crab body weight, respectively, while the safe concentration was 82.5 mg OTC kg(-1) crab weight. We suggest that OTC has potential as a highly effective inhibitor of Spiroplasma pathogens in aquatic animals and has been proven to be a potent, safe and low cost cure for TD. This represents a novel use of OTC in the therapeutic treatment of an aquacultural disease caused by a Spiroplasma pathogen. PMID- 19565703 TI - Huffmanela sp. (Nematoda: Trichosomoididae) muscular parasite from Trisopterus luscus captured off the Portuguese coast. AB - Four samples of Trisopterus luscus exhibiting unusual black spots on their musculature were obtained from catches made on different days and were rejected as unsuitable for consumption. Bipolar eggs somewhat resembling those of the nematode genus Huffmanela (Nematoda: Trichosomoididae) were identified as the cause of these black spots. Microscopic analysis revealed the presence of dark shelled trichinelloid embryonated eggs dispersed throughout the muscle tissue, 73 to 94 x 40 to 59 microm in size, with large polar plugs 14 to 22 microm wide. However, the morphology and distribution of the observed eggs were not compatible with any previously described Huffmanela species. Intracellular nematodes were also observed in muscle cells. In 2 samples, sectioned nematode specimens were observed in other locations as well: the lymphatic vessel of intestine submucosa and the dermal lymphatic vessel. This could be due to nematode migration inside the infected fish. The presumed identification of Huffmanela sp. in a new host (Trisopterus luscus), and in a new location (Atlantic Ocean) supports the belief that these parasites could be widely distributed, particularly in marine fishes. In view of these factors, together with the observed egg morphology, it is possible that these specimens represent a new, unidentified Huffmanela species. PMID- 19565704 TI - [Initial RAPD analysis of Isatis indigotica somatic clones' resistance to root rot disease]. AB - OBJECTIVE: To make initial identification on Isatis Indigotica Somatic Clones' resistant mutant. METHODS: The resistant plants were selected under the treatment of root-rot disease crude toxin. The DNA of the same clones and their parents were analysed by RAPD technology. RESULTS: We got initial resistance of the plants and selected 12 primers of which most were identical but the less were different. The genetic background of the somatic clones was identical but the individuals were mutated on the DNA level. CONCLUSION: It provides experimental foundation for further study on separating resistant mutant and resistant gene. PMID- 19565705 TI - [Study on the autointoxication of aquatic extracts from different parts of Coptis chinensis]. AB - OBJECTIVE: To study the autointoxication of aquatic extracts from different parts of Coptis chinensis. METHODS: By measuring the effects on seed germination, seedling growth and adult-plants' growth, the autointoxications of root exudates, rhizome extracts, plant residue decomposition products and aqueous extracts from rhizosphere soil were studied. RESULTS: The aquatic extracts from different parts of C. chinensis could inhibit the nutritional growth stage of C. chinensis. CONCLUSIONS: Autointoxication is one of the factors leading to continuous cropping obstacle, and this study is instructive to the management and application of C. chinensis in agricultural productivity. PMID- 19565706 TI - [Study on quality standard of Mucuna pruriens var. utilis]. AB - OBJECTIVE: To provide scientific basis for the utilization and development of Mucuna pruriens var. utilis by establishing its quality control standard. METHODS: The bioactive constituents were analyzed by TLC and HPLC. Moisture, ash and the extracts of Mucuna pruriens var. utilis were all determined. RESULTS: The TLC spots of levodopa had similar color with the control group at the same position. The results of HPLC quantitative analysis showed that the linear range of levodopa was 26.45 to approximately 132.25 microg/mL, r = 0.9992, and the average recovery rate was 103.8%, RSD = 1.85%. CONCLUSIONS: This method is convenient, accurate, reliable with good reproducibility, so it can be used to establish quality standard for the medicinal material. PMID- 19565707 TI - [Naphtha analysis of different processed products of Rhizoma Curcumae from Guangxi by GC-MS]. AB - OBJECTIVE: To approach the naphtha analysis of different processed products of Rhizoma Curcumae from Guangxi. METHODS: The naphtha was extracted by steam distillation. The relative content of every compound was determined with area normalization method and the structures were elucidated by GC-MS technique. RESULTS: 33 kinds of chemical compositions were identified in the naphtha of the six samples,the main contents were Cyclohexane, 1-ethenyl-1-methyl-2,4-bis (1 methylethenyl)-, 5-Benzofuranacetic acid, 6-ethenyl-4,5,6,7-terahydro-3, 6 dimethyl-alpha. -methylene-,methyl ester, and so on. CONCLUSIONS: Six samples of Rhizoma Curcumae are significantly different in quality,and the naphtha's content of the sample processing with vinegar is the highest. PMID- 19565708 TI - [Microscopical identification and hierarchical cluster analysis of seven kinds of pilose antler velvet]. AB - OBJECTIVE: According to the microscopical features of seven kinds of pilose antler velvet, to identify various kinds of pilose antler. METHODS: To identify seven kinds of pilose antler velvet by bio-microscope, then the classification was carried out by hierarchical cluster analysis. RESULTS: The microscopic identification of seven kinds of pilose antler velvet indicators were made and effectively classified. CONCLUSION: This method is useful in the identification of pilose antler,and provides reference for its qualitative evaluation. PMID- 19565709 TI - The chemical constituents from leaves of Acer saccharum. AB - OBJECTIVE: To study the chemical constituents of leaves of Acer saccharum. METHODS: The leaves of Acer saccharum were extracted by ethanol. The concentrated material was partitioned by petroleum ether, ethyl acetate, and n-butanol. After extracted by ethyl acetate, the extract was isolated and purified by silica gel column chromatography and recrystallization. The compound structures were identified on the basis of spectral data and chemical methods. RESULTS: Seven compounds were isolated from the leaves of Acer saccharum. Their compound structures were identified as 3-keton-ursane,3beta-hydroxy-12-olean-12-en and 5 en-7-hydroxy-sitosterol. CONCLUSION: All the three compounds identified are isolated from this genus for the first time. PMID- 19565710 TI - [Studies on the chemical constituents from Xinjiang Punica granatum]. AB - OBJECTIVE: To study the chemical constituents from Punica granatum. METHODS: The chemical constituents were isolated by column chromatography on silica gel and MCI-gel CHP 20P, Sephadex LH-20. Their structures were elucidated on the basis of spectral analysis and physical constants. RESULTS: Ten compounds were isolated and elucidated as: gallic acid (1), methyl gallate (2), ellagic acid (3), (+) catechin (4), isoquerecitrin (5), D-mannitol (6), ursolic acid (7), Oleanolic acid (8), beta-Sitosterol (9) and Daucosterol (10). CONCLUSION: Compounds 1-10 are isolated from this plant for the first time. PMID- 19565711 TI - [Studies on the chemical constituents of Chelonopsis albiflora]. AB - OBJECTIVE: To study the chemical constituents of Chelonopsis albiflora. METHODS: The compounds were isolated and purified by various column chromatographic techniques and their structures were elucidated on the basis of spectral analysis (EI-MS), 1H-NMR, 13C-NMR. RESULTS: Seven compounds were isolated and identified as 1beta-hydroxylupenol (1), lupenol (2), beta-sitosterol (3), 13-epi-torulosal (4), 19-acetoxy-13-hydroxylabada-8 (17), 14-diene (5), (E)-p-methyl cinnamic acid (6) and vanillic acid (7) . CONCLUSION: All compounds are isolated for the first time from this genus. PMID- 19565712 TI - [Studies on the chemical constituents of the aerial part of Bupleurum chinense (II)]. AB - OBJECTIVE: To study the chemical constituents of the aerial part of Bupleurum chinense. METHODS: Dried aerial parts of Bupleurum chinense were extracted with EtOH, the chemical constituents in the extract were isolated and their structures were identified by physical and chemical properties and spectral analysis. RESULTS: Five compounds were obtained and identified as 1-O-caffeoylglycerol (1), esculetin (2), scopoletin (3), kaempferol-3-O-beta-D-rutinoside (4) and alpha spinastero (5). CONCLUSION: Compounds 1-3 are obtained from Bupleurum chinense for the first time. PMID- 19565713 TI - [Studies on the chemical constituents of MEICHA (II)]. AB - OBJECTIVE: To study the chemical constituents of MEICHA. METHODS: Chromatography and spectroscopic analysis were employed to isolate and elucidate the chemical constituents in the plant. RESULTS: Seven compounds were obtained and identified as beta3-sitosterol (I), oleanolic acid (II), quercetin (III), Taxifolin (IV), Catechin (V), Querceetin-3-O-alpha-L-rhamnoside (VI) and Gallic acid (VII). CONCLUSION: Seven compounds are isolated from this plant for the first time. PMID- 19565714 TI - [Studies on the chemical constituents of Nerviliae fordii]. AB - OBJECTIVE: To study the chemical constituents of Nerviliae fordii. METHODS: The compounds were isolated by silica gel and sephadex LH-20 column chromatography. Their structures were elucidated on the basis of physico-chemical properties and spectral datas. RESULTS: Five compounds were isolated and identified as rhamnocitrin (1), rhamnazin (2), rhamnocitrin 3-O-beta-D-glucopyranoside (3), rhamnocitrin 4'-O-beta-D-glucopyranoside (4) and 4-hydroxybenzonic acid (5). CONCLUSION: Compound 3, 4, and 5 are isolated from this plant for the first time. PMID- 19565715 TI - [Quantitative determination of 1-deoxynojirimycin in mulberry leaves by high performance liquid chromatographic-tandem mass/mass spectrometry]. AB - OBJECTIVE: To develop a high-performance liquid chromatographic-tandem mass/mass spectrometric method to determine the concentration of 1-deoxynojirimycin (1-DNJ) in mulberry leaves. METHODS: 1-deoxynojirimycin was separated on an SHIMADZU HRC NH2 column with the mobile phase consisting of acetonitrile and 0.1% formic acid. The mass spectrometric system equipped with a atmospheric pressure chemical ionization (APCI) interface was operated in Multiple Reaction Monitoring (MRM) mode. RESULTS: The retention time of 1-deoxynojirimycin was 2.87 min, and the calibration curve was linear over a concentration range from 482 microg/L to 2410 microg/L, the average recovery was 95.8%. The detection limit was 53.6 microg/L. CONCLUSION: The method is selective and sensitive for determining 1 deoxynojirimycin in mulberry leaves. PMID- 19565716 TI - [Protective effect of Buyanghuanwu Decoction on myocardial ischemia induced by isoproterenol in rats]. AB - OBJECTIVE: To study the protective effect of Buyanghuanwu Decoction on myocardial ischemia induced by isoproterenol in rats. METHODS: Buyanghuanwu Decoction was given in different dose and the rat model of myocardial ischemia was established by peritoneal injection of isoproterenol. The expression of CD40 in whole blood was detected by flow cytometry,and the expression of CD40L in myocardial tissues was detected by immunohistochemistry. The activities of lactate dehydrogenase (LDH), creatine kinase (CK) and aspartate aminotransferase (AST) in blood serum were detected by biochemistry detector. RESULTS: Compared with the model group, Buyanghuanwu Decoction in high and middle dose significantly inhibited the expression of CD40 in blood serum and CD40L in myocardial tissues (P < 0.01), and obviously decreased the activities of LDH, CK and AST in blood serum (P < 0.01). CONCLUSION: Buyanghuanwu Decoction has a protective effect on myocardial ischemia induced by isoproterenol in rats, and it may be relevant to the decrease of the expression of CD40-CD40L and the activities of myocardial enzymes. PMID- 19565717 TI - [Effect of berberine on the differentiation and apoptosis of K562 cell line]. AB - OBJECTIVE: To study the effect of Berberine on proliferation, differentiation and apoptosis of K562 cells for supplying the theoretical evidence on the clinical application of Coptis chinensis to cure chronic myeloid leukemia. METHODS: The proliferation-inhibiting capability of K562 cells was investigated by MTT assay and colony conform test. The apoptosis effect of Berberine on K562 cells were analyzed by Wright's-Giemsa staining; DNA fragmentation was performed by agarose gel electrophoresis. The cell cycle distribution and the cell surface marker cluster of differentiation were determined by flow cytometry. RESULTS: Berberine effectively inhibited the proliferation of K562 cells in a dose and time dependent manner. In addition, when combined with different concentrations of Ara C, the cells' viability percentage was prominently higher than only Berberine used. Treated with Berberine for 48 hours, the cells could be induced differentiation towards erythrocyte, granulocyte and megakaryocyte and with the treated time extending, the percentage of apoptosis cells gradually increased. CONCLUSIONS: Berberine can efficiently inhibit the proliferation of K562 cells, maybe by the way of blocking cells at the stage of G0/G1 and (or) G2, then leading to its apoptosis and differentiation. PMID- 19565718 TI - [Study of the property of lipids reducing of curcumin on hyperlipidemia mice after fermented by Monascus purureus]. AB - OBJECTIVE: To study the lipids reducing property of curcumin on Hyperlipidemia mice after fermented by Monascus purureus. METHODS: The stain Monascus purureus was used for microbial transformation, and both substrate control and strain control were set. The mice were reared with high lipid and cholesterol feed for 15d to establish the Hyperlipidemia models. The models were treated with fermented curcumin in 500 mg/kg, 200 mg/kg, 100 mg/kg, substrate control and strain control in 500 mg/kg. Positive and Normal group were treated with natural saline. The general situation was observed and the changes of TG, TC, HDL-C levels in serum and liver were tested after 10 d. RESULTS: Fermented curcumin could significantly reduce the serum TC, TG of Hyperlipidemia mice all in high, middle and low doses. Serum TC was reduced by 38.7%, 34.5%, 32.7% and TG was reduced by 38.3%, 28.6%, 30.1%, respectively while substrate control and strain control had no effect. Fermented curcumin also could reduce the TC, TG in liver but no effect of curcumin substrate at the same dose. CONCLUSION: The property of lipids reducing of curcumin is significantly enhanced after fermentation. PMID- 19565719 TI - [Antiviral effects of an effective section of a prescription of traditional Chinese medicine on influenza virus A in vitro]. AB - OBJECTIVE: To study the inhibitive effects of an effective section of a prescription of traditional Chinese medicine (TCM-ES) on influenza virus A FM1 strain in vitro. METHODS: The experiments were performed by microcytopathic inhibiting-assay, Neutral Red stain and inhibiting plaque-forming units (PFU) test on MDCK cell strain. By means of observing the cytopathic effects (CPE), measuring the absorbance [D(lambda)] and counting the PFU, according to Reed Muench assay, the TCM-ES's effective dosage of 50 percentage (EC50) and treatment index (TI) to FM1 were calculated. The inhibiting dose of 50 percentage of PFU (IC50) was also figured up. RESULTS: By CPE assay, TCM-ES'S EC50, MTC and TI to 100TCID50 FM1 strain infection were (300 +/- 18.3) mg/L, (75 +/- 6.8) mg/L and (7.1 +/- 0.7), respectively; Whereas, ribavirin's EC50, MTC and TI was (52.3 +/- 10.1) mg/L, (25 +/- 4.1) mg/L and (20.8 +/- 5.1), respectively. By Neutral Red stain assay,TCM-ES's IC50 and TI was (285.0 +/- 19.2) mg/L and (7.2 +/- 0.6), respectively; whereas ribavirin's IC50 and TI was (45.3 +/- 4. 9) mg/L and (21.2 +/- 3.1), respectively. By reducing PFU assay, the IC50 of TCM-ES and ribavirin was 300 mg/L and 50 mg/L, respectively. All the results above were almost consistent with each other (P > 0.05). CONCLUSIONS: TCM-ES assumes antiviral action on IFV-FM1 strain in a certain degree in vitro and can rebel intracellular virus. But it is worse than the positive control medicine of ribavirin and is worthy of further study. PMID- 19565720 TI - [Protective effect of puerarin on the secondary spinal cord injury in rats]. AB - OBJECTIVE: To research the protective effect of puerarin on secondary spinal cord injury (SCI) in rats. METHODS: After the models of SCI were established by improved Allen's method on adult male SD rats, SOD, MDA, Bcl-2 and Bax gene protein expression between puerarin group and model group were compared after 1, 6, 12, 24 and 48 h. RESULTS: Puerarin could significantly enhance the activity of SOD and reduce the content of MDA, increase the expression of Bcl-2 gene protein products and decrease Bax gene protein product. CONCLUSIONS: Puerarin can increase the activity of SOD, reduce the content of MDA, promote the expession of Bcl-2 and restrain the expression of Bax in the early spinal cord injury. It has protective effect on the secondary spinal cord injury. PMID- 19565721 TI - [Protective effect of polysaccharides from sea cucumber on glu-induced neurotoxicity in PC12 cells]. AB - OBJECTIVE: To investigate the protective effect of polysaccharides from sea cucumber (HS) on Glu-induced neurotoxicity in PC12 cells. METHODS: The toxicity model was established by treating PC12 cells with Glu in vitro. Cell viability was assayed with MTT method. The vitality of antioxidant enzymes (SOD) and the level of malondialdehyde (MDA) and lactic dehydrogenase (LDH) were evaluated by chromatometry. RESULTS: The vitality of SOD increased and the levels of MDA and LDH significantly decreased in PC12 cells of HS group compared with the model group. HS in different concentration showed protective effects as it increased the vitality of SOD and decreased the content of MDA and LDH in a dose-dependent manner. CONCLUSION: Polysaccharide from sea cucumber can protect PC12 cells from the injury induced by Glu. PMID- 19565722 TI - [Effect of Ning Wei Kang Recipe extract on gastrointestinal motor function]. AB - OBJECTIVE: To study the effect of Ning Wei Kang Recipe extract on gastrointestinal motor function. METHODS: The effects of Ning Wei Kang Recipe extract on gastrointestinal motor function were studied in mice, rabbits, and isolated mesenterial intestines of guinea pig according to its effect on acetylcholine or barium chloride-induced contraction. RESULTS: Ning Wei Kang Recipe extract could significantly decrease the propellent movement of medicine charcoal in the mesenterial intestine of mice (P < 0.01), inhibit the range and speed of mesenterial intestine of rabbits (P < 0.05, or P < 0.01), relax the contration of isolated mesenterial intestines of guinea pig induced by BaCl2 (P < 0.05). But it could not decrease the contration of isolated mesenterial intestines of guinea pig induced by acetylcholine (P< 0.05). CONCLUSIONS: Ning Wei Kang Recipe extract can significantly inhibit the gastrointestinal motor function. It may be related to blocking positive ion-Ba2+ into gastrointestinal smooth muscle cell. PMID- 19565723 TI - [Therapeutic effects of traditional Chinese herbal formula with Smilax glabra on mice's lead poisoning]. AB - OBJECTIVE: To investigate the influence of traditional Chinese herbal formula with Smilax glabra on lead expelling and oxidative damage in lead-poisoned mice. METHODS: Mice were received intraperitoneal injections of lead acetate to establish lead-poisoned models. The concentration of lead, zinc and copper in blood, brain, liver, kidney and bone were determined. The lipid peroxidation of liver was systematically evaluated. RESULTS: The traditional Chinese herbal formula had no influence on body weight and organ weight/body weight ratio of lead-poisoned mice. The traditional Chinese herbal formula might significantly reduce the concentration of lead in blood, rain, liver, kidney and bone, and the rate were 74%, 48%, 56%, 51% and 44%, but had no influence on zinc's and copper' s concentration. The decreasing rate of MDA levels in liver by the traditional Chinese herbal formula was 31%. In addition, the increase rate of GSH levels and the activities of GSH-Px and SOD in liver by traditional Chinese herbal formula were 63% ,76% and 66%, respectively. CONCLUSIONS: The traditional Chinese herbal formula with Smilax glabra can significantly advance the excretion of lead and protect liver against oxidative damage induced by lead. PMID- 19565724 TI - [Preparation and characterization of curcumin loaded gelatin microspheres for lung targeting]. AB - OBJECTIVE: To study the preparation technology and physicochemical characterization of curcumin loaded gelatin microspheres for lung targeting. METHODS: Curcumin loaded gelatin microspheres for lung targeting were prepared by the emulsion crosslinking method, and the preparation technology was optimized by orthogonal experimental design with biodegradable gelatin as the carrier, liquid paraffin as the oil phase, span 80 as the emulsifier. RESULTS: The optimal curcumin loaded gelatin microspheres were global with smooth surface, 6.15% for the drug loading, 75.5% for the encapsulation efficiency. 86.6% of microspheres size was in the range of 5-30 microm. The release test in vitro showed that 50% of curcumin could release from gelatin microspheres in 22 h and 77% in 48 h. CONCLUSIONS: The preparation technology is good and stable, and the obtained microspheres can control the release of curcumin. PMID- 19565725 TI - [Study on the optimal extracion craft of Xiaochuang gel by central composite design-response surface methodology]. AB - OBJECTIVE: To optimize the extraction craft of Xiaochuang gel by central composite design-response surface methodology. METHODS: The main influential factors of extraction effect included the quantity of ethanol, the concentration of ethanol and the extraction time. The conditions of the extraction were evaluated by the yield of the extracts and the total content of rheum emodin and chrysophanol. The optimal extraction craft of Xiaochuang gel was chosen by central composite design-response surface methodology. RESULTS: The optimal conditions were extracting two times, two hours each time with ten volume multiples of 60% ethanol. CONCLUSIONS: It is simple, convenient and highly predictive to optimize the extraction craft of Xiaochuang gel by central composite design-response surface methodology. PMID- 19565726 TI - [Study on the optimal extaction of total flavonoids from Tagetes erecta by central composite design-response surface methodology]. AB - OBJECTIVE: To optimize the process of extracting total flavonoids from Tagetes erecta. METHODS: The influential factors were extraction temperature, ethanol concentration, reflux time and solvent volume fold. The evaluating indicator was the extraction rate of total flavonoids from Tagetes erecta. The central composite design-response surface methodology was used to optimize the process and the prediction was carried out. RESULTS: The optimum conditions of extraction were 80% ethanol, 2.5 hours for reflux, 35 volume folds of solvent and 70 degrees C. CONCLUSION: It shows that the optimum model is simple and highly predictive. PMID- 19565727 TI - [Effect on bioactive components of Xuesaitong injection in the process of ultra filter]. AB - OBJECTIVE: To study the effect on bioactive components of Xuesaitong injection in the process of ultra-filter by orthogonal test. METHODS: The change of the contents of active components was examined by HPLC method, using notoginsennoside R1 , ginsennoside Rg1, ginsennoside Rb1 and ginsennoside Rd as the mark components. The effect of the transmissivity of active components was studied with four different membrane parameters (such as pore diameter of membrane, operational pressure, the temperature and the concentration of the officinal liquid) by orthogonal test. RESULTS: In the process of ultra-filter the transmissivity of notoginsennoside R, and ginsennoside Rg, had the same characteristics. Both of the average transmissivities of these two components were in the range of 45% to 95%. The concentration of the officinal liquid showed significant influence. The transmissivities of ginsennoside Rb, and ginsennoside Rd in the process of ultra-filiter had the same characteristic. The average transmissivities of these two components were in the range of 2 8%-62% and 1.8% 45%, respectively. The pore diameter of membrane and the temperature showed significant influence. The concentration of the officinal liquid also showed significant influence on the transmissivity of ginsennoside Rd. CONCLUSION: This study provides an external direction for the preparation of Xuesaitong injection. PMID- 19565728 TI - [Preparation of compound PuHuang granules and its clinical efficacy]. AB - OBJECTIVE: To prepare Compound PuHuang Granules and observe its clinical efficacy on cerebral infarction. METHODS: The Compound PuHuang Granules were prepared and the identification was performed by TLC. A total of 328 patients with cerebral infarction were randomly divided into two groups and observed. 197 cases in the trial group were administered with Compound PuHuang Granules for 2 weeks, and 131 cases in the control group were administered with NaoXueShuan Tablets for 2 weeks as well. RESULTS: The method of identification was proved to be specific and reproductive. The effective rates of the trial group and the control group on vertigo, hemiplegia, numbness of extremities were 88.71%, 82.46%, 84.38% and 61.11%, 62.07%, 66.67%, respectively, which showed significances between the two groups (P < 0.05). But the effective rates of two groups on language disorder showed no signifcance. CONCLUSIONS: The Compound PuHuang Granules is feasible in preparation and it has obvious efficacy, therefore, it would be useful in the clinical applications. PMID- 19565729 TI - [Acupuncture at points of the liver and gallbladder meridians for treatment of migraine: a multi-center randomized and controlled study]. AB - OBJECTIVE: To investigate the therapeutic effect and safety of acupuncture at points of The Liver and Gallbladder Meridians for treatment of migraine. METHODS: Multi-central, randomized and controlled trial was used and 253 cases of migraine were divided into an acupuncture group and a western medicine group. The acupuncture group was treated with acupuncture at points of The Liver and Gallbladder Channels with Taichong (LR 3), Yang-lingquan (GB 34), Fengchi (GB 20), Ququan (LR 8) selected as main points, and the western medicine group with oral administration of Flunarizine tablets for 4 therapeutic courses. The total therapeutic effects 3 and 6 months after the treatment, the scores of various symptoms of migraine before and after treatment, and the stability of therapeutic effect in one-year following-up survey were observed in the two groups. RESULTS: After treatment, mean times and duration of the headache attack were significantly improved in the two groups (all P < 0.01) with the acupuncture group better than the western medicine group (P < 0.05). The total effective rates for stopping pain after treatment, 3 months and 6 months after treatment in the acupuncture group were 93.0%, 93.0% and 87.7%, respectively, which were better than 85.6%, 86.5% and 69.2% in the western medication group (all P < 0.01). One year later, the stability of the therapeutic effect in the acupuncture group was better than that in the western medicine group (P < 0.05); the adverse reaction and the compliance in the acupuncture group were significantly superior to those in the western medicine group. CONCLUSION: Acupuncture at points of The Liver and Gallbladder Meridians for treatment of migraine is safe, effective, and with stable long-term therapeutic effect. PMID- 19565731 TI - [Clinical observation on acupuncture combined with Yizhi Jiannao granules for treatment of Alzheimer's disease]. AB - OBJECTIVE: To observe clinical therapeutic effect of acupuncture combined with Yizhi Jiannao Granules for treatment of Alzheimer's disease and its effects on intelligence, daily life and social activity ability. METHODS: Eighty-four cases were randomly divided into 3 groups, 28 cases in each group. The combined acupuncture and medication group was treated with acupuncture at Baihui (GV 20), Sishencong (EX-HN 1), Dazhui (GV 14), Guanyuan (CV 4), etc. and oral administration of Yizhi Jiannao Granules; the Chinese herb group was treated with Yizhi Jiannao Granules, and the western medicine group with oral administration of Aricept. The scores for the Mini-Mental State Examination (MMSE), Ability of Daily Life (ADL) and the therapeutic effects were assessed and compared before treatment and after treatment for 12 weeks among the groups. RESULTS: After treatment, the scores for MMSE and ADL were improved in the combined acupuncture and medication group, the Chinese herb group and the western medicine group, which were better in the combined acupuncture and medication group (P < 0.05). The total effective rate of 85.7% in the combined acupuncture and medication group was better than 71.4% in the Chinese herb group and 67.9% in the western medicine group. CONCLUSION: Acupuncture combined with Yizhi Jiannao Granules has a significant therapeutic effect on Alzheimer's disease, which is better than that of Yizhi Jiannao Granules or Aricept. PMID- 19565730 TI - [Observation on therapeutic effect of acupoint application on dysmenorrhea of excess syndrome and effect on prostaglandins]. AB - OBJECTIVE: To observe therapeutic effect of acupoint application on dysmenorrhea of excess syndrome and its effect on prostaglandins. METHODS: The patients with primary dysmenorrhea of excess syndrome were randomly divided into an application group and a medication group. The application group of 31 cases were treated with application of Chinese medicine composed of Zhinanxing (Rhizoma Arisaematis), Sanleng (Rhizoma Sparganii), Ezhu (Rhizoma Zedoariae), etc. at Zhongji (CV 3), Guanyuan (CV 4), Qihai (CV 6); and the medication group of 30 cases were treated with oral administration of Tianqi Tongjing Capsules. The scores for the pain degrees and the duration of dysmenorrhea were observed and prostaglandin F2alpha (PGF2alpha) and prostaglandin E2 (PGE2) contents in peripheral blood were determined before and after treatment. RESULTS: The total effective rate of 93.5% in the application group was significantly better than 73.3% in the medication group (P < 0.05); after treatment, the scores of the symptoms were significantly decreased in the two groups (both P < 0.01), with more obvious improvement in the application group than the medication group (P < 0.01). The PGE2 content was significantly increased, and the PGF2alpha content and PGF2alpha/PGE2 were significantly decreased in the application group (all P < 0.01). CONCLUSION: Acupoint application has a better therapeutic effect on dysmenorrhea of excess syndrome and has benign regulative action on synthesis of prostaglandins in the patients with primary dysmenorrhea. PMID- 19565732 TI - [Observation on therapeutic effect of acupoint application of Chinese medicine on bronchial asthma]. AB - OBJECTIVE: To compare therapeutic effects of acupoint application of Chinese medicine and routine acupuncture on bronchial asthma. METHODS: One hundred cases were randomly divided into an observation group and a control group, 50 cases in each group. The observation group was treated with application of Chinese medicine at Dazhui (GV 14), Feishu (BL 13), Gaohuang (BL 43), Danzhong (CV 17), Qihai (CV 6) in the dog days; and the control group was treated with acupuncture at Zhongfu (LU 1), Tiantu (CV 22), Danzhong (CV 17), Dingchuan (EX-B 1), Fenglong (ST 40). The long-term therapeutic effects were observed. RESULTS: The total effective rate was 94.0% in the observation group and 76.0% in the control group with a significant difference between the two groups (P < 0.05). CONCLUSION: The therapeutic effect of acupoint application of Chinese medicine in the dog days on bronchial asthma is significantly superior to that of routine acupuncture. PMID- 19565733 TI - [Observation on therapeutic effect of warm needle moxibustion on chronic non bacterial prostatitis]. AB - OBJECTIVE: To observe the therapeutic effect of warm needle moxibustion on chronic non-bacterial prostatitis. METHODS: One hundred and twenty-five cases were randomly divided into a warm needle moxibustion group (n = 42), an acupuncture group (n = 41) and a western medicine group (n = 42). The warm needle moxibustion group was treated with acupuncture at Shenshu (BL 23), Ganshu (BL 18), Zhibian (BL 54) with no retention of the needles, and warm needle moxibustion on Guanyuan (CV 4), Zhongji (CV 3), Yinlingquan (SP 9), Sanyin-jiao (SP 6), once daily; the acupuncture group with acupuncture at the same acupoints as the warm needle moxibus tion group with no moxibustion; and the western medicine group with oral administration of Prostat Tablets, one tablet each time, twice daily. The therapeutic effects and scores of chronic prostatitis symptom indexes (NIH-CPSI) were compared. RESULTS: The total effective rate was 88.10% in the warm needle moxibustion group, 63.41% in the acupuncture group and 66.67% in the western medicine group, the warm needle moxibustion group being better than the acupuncture group and the western medicine group (both P < 0.05). After treatment, the scores of NIH-CPSI was 11.92 +/- 7.11 in the warm needle moxibustion group, 16.08 +/- 6.83 in the acupuncture group and 15.66 +/- 5.88 in the western medicine group, with significant decreases in the 3 groups as compared with those before treatment (all P < 0.01), and after treatment, the score in the warm needle moxibustion group was significant different to that of the acupuncture group or the western medicine group (both P < 0.01). CONCLUSION: The warm needle moxibustion has a good therapeutic effect on chronic non bacterial prostatitis. PMID- 19565734 TI - [Acupuncture combined with magnetic therapy for treatment of temple-jaw joint dysfunction]. AB - OBJECTIVE: To compare clinical therapeutic effects of acupuncture combined with magnetic therapy and simple magnetic therapy on temple-jaw joint dysfunction. METHODS: Eighty-two cases were randomly divided into an observation group (n = 52) and a control group (n = 30). The observation group was treated with acupuncture at Xiaguan (ST 7), Jiache (ST 6), Hegu (LI 4), etc. and AL-2 low frequency electromagnetic comprehensive treatment instrument; the control group was treated with AL-2 low frequency electromagnetic comprehensive treatment instrument. RESULTS: The cured and markedly effective rate of 90.4% in the observation group was significantly better than 66.7% in the control group (P < 0.01), and the total effective rate of 98.1% in the observation group was significantly better than 86.7% in the control group (P < 0.05). CONCLUSION: The therapeutic effect of acupuncture combined with magnetic therapy is significantly better than that of the simple magnetic therapy on temple-jaw joint dysfunction. PMID- 19565735 TI - [Analysis on therapeutic effect of bird-pecking moxibustion combined with western medicine for treatment of infantile hand, foot and mouth disease]. AB - OBJECTIVE: To observe therapeutic effects of bird-pecking moxibustion in children of hand, foot and mouth disease, and to analyze the mechanism. METHODS: Seventy five children of hand, foot and mouth disease were randomly divided into 3 groups, a combined moxibustion and medicine group (n = 22), a Chinese medicine group (n = 29), and a western medicine group (n = 24). The combined moxibustion and medicine group was treated with bird-pecking moxibustion combined with routine western medicine, the Chinese medicine group with oral administration of Chinese medicine and routine western medical therapy, and the western medicine group with routine western medicine. After treatment of 7 days, the therapeutic effects on skin rash, oral herpes, constipation or loose stool, dyspepsia and anorexia, etc. were comprehensively assessed, and their therapeutic effects were compared. RESULTS: The total effective rate was 95.5% in the combined moxibustion and medicine group, 86.2% in the Chinese medicine group, and 83.3% in the western medicine group, the former being significantly better than those of the other two groups (both P < 0.05). The combined moxibustion and medicine group was significantly better than the other two groups in the relieving time of skin rash, oral herpes, constipation or loose stool, dyspepsia and anorexia, etc. (all P < 0.05) CONCLUSION: The combined moxibustion and medication can effectively improve symptoms of the digestive tract, shorten duration of disease, reduce pain in the patient with hand, foot and mouth disease. PMID- 19565736 TI - [Pricking blood therapy combined with ultraviolet irradiation for treatment of acute herpes zoster]. AB - OBJECTIVE: To evaluate clinical therapeutic effect and the safety of pricking blood therapy combined with ultraviolet irradiation for treatment of acute herpes zoster. METHODS: One hundred and thirty cases were randomly divided into an observation group and a control group, 65 cases in each group. The observation group was treated with pricking blood therapy combined with ultraviolet irradiation. Firstly, the affected parts were heavily taped with a plum-blossom needle and then cupping. After the cup was removed, with the body surface dividing field method, ultraviolet irradiation was given at the skin injury area and the nerve root area corresponding to paraspinal vertebra, and the control group was treated with Aciclovir and other western medicine. Seven days constituted one course. Their therapeutic effects and adverse reactions were observed. RESULTS: After treatment of 7 days, the cured rate of 76.9% and the total effective rate of 90.8% in the observation group were significantly higher than 38.5% and 66.2% in the control group, respectively (both P < 0.01); the incidence rate of post herpetic neuralgia of 3.1% in the observation group was significantly lower than 12.3% in the control group (P < 0.05); after treatment, the scores for pain, rash and sleep decreased significantly in the two groups (all P < 0.01), more significantly decreased in the observation group than in the control group (P < 0.01 or P < 0.05); the pain-relieving time, herpes-stopping time, scab-forming time and the cured time in the cured patients of the observation group were significantly shorter than those in the control group (P < 0.01 or P < 0.05). CONCLUSION: The pricking blood therapy combined with ultraviolet irradiation has rapid therapeutic effect, effectively shortens duration of illness, decreases the incidence rate of post herpetic neuralgia and it is a safe remedy for treatment of herpes zoster. PMID- 19565737 TI - [Study on needling depth and direction from different acupoints to sphenopalatine ganglion]. AB - OBJECTIVE: To observe and survey the location of Xiaguan (ST 7), "Die'e" and Quanliao (SI 18) on the surface, and the needling depth and direction from the 3 points to sphenopalatine ganglion. METHODS: Fifteen corpses (30 sides) of adult male were fixed by 10% formalin. The lateral areas of face were dissected from the surface to the deep on the 3 acupoints: the electric drill with the kirschner wire punctured towards the sphenopalatine ganglion and extended to the contralateral areas according to different directions of puncturing sphenopalatine ganglion from the 3 acupoints. The corresponding puncturing points of the 3 acupoints were measured by the coordinate location method. RESULTS: (1) Surface location: the distance between Quanliao (SI 18) and "Die'e" was 21 mm and the distance between Xiaguan (ST 7) and "Die'e" was 17 mm; (2) Inserting depth of each point to sphenopalatine ganglion: the depths of Xiaguan (ST 7), "Die'e" and Quanliao (SI 18) were 49.9 mm, 46.9 mm and 46.6 mm, respectively; (3) The coordinate location of the corresponding puncturing points: the puncturing direction of Xiaguan (ST 7) was anterointernal upper corresponding to the area of connecting center between contralateral Taiyang (EX-HN 5) and Tongziliao (GB 1), the distance between the corresponding inserting point of Xiaguan (ST 7) and Sizhukong (TE 23) was 17.6 mm; the puncturing direction of "Die'e" point was posterointernal upper, and the horizontal distance from the corresponding puncture point to the zygomatic arch was 33 mm and the vertical distance from the corresponding puncture point to the eyes' outer canthus was 42 mm; the puncturing direction of Quanliao (SI 18) was posteriointernal upper and the distance between the corresponding inserting point and the area of contralateral parietal tuber, the distance between the corresponding inserting point of Quanliao (SI 18) and the connecting line of bilateral external acoustic pore was 28 mm, the distance between the corresponding inserting point of Quan-liao (SI 18) and the medial line of the head was 62 mm. CONCLUSION: Understanding the surface location, inserting depths and the general puncturing directions of the 3 points can provide basis for puncturing the sphenopalatine ganglion in clinical practice. PMID- 19565738 TI - [On structure characteristics and mechanisms of channels and collaterals]. AB - Recognition of traditional Chinese medicine for the structures of channels and collaterals is possibly related with the tactics of human early general medical study, characterized by "inner", sub-micro, entirety, etc. The mechanisms are possibly involved in direct communication pathway of the cell junction (specially keeping the relevant cellular direct communication relation in individual development), the extra-cellular matrix stress network system and its biophysical message transmission--force--meridian signs transformation--inter-cellular massage integration, and "generalization" of the structures and activities of channels and collaterals in acupuncture serum phenomenon, etc. Thus, the hypothesis of multi-factors about channel structures-mechanisms is proposed. PMID- 19565739 TI - [Professor Feng Run-Shen's essential experience in penetration needling method]. AB - Professor Feng Run-Shen is engaged in medicine for more than 60 years. He pays attention to medical ethics and has perfect medical skill. He energetically advocates combination of acupuncture with medication and stresses the concept of viewing the situation as a whole in selection of acupoints and treatment, particularly, clinical application of point properties. Clinically, he is accomplished in penetration needling, for which one needle acts on two or more points, enlarging the range of needling sensation, so it has very good therapeutic effects on many diseases. In the paper, the case samples about penetration needling in his clinical practice are summarized and introduced. PMID- 19565740 TI - [Effects of acupuncture on the expression of brain-derived neurotrophic factor in the ovariectomized rat fracture model]. AB - OBJECTIVE: To observe the effect of acupuncture on fracture in the ovariectomized rat and the mechanism. METHODS: Sixty SD female rats were randomly divided into 4 groups: normal group (group A), model group (group B), acupuncture group (group C) and Nilestriol group (group D). In all the groups, except the group A which received sham operation, the rats were overiectomized for preparing the osteoporosis model. Three months after the ovariectomy, the left femurs of the rats were closely fractured. Then, the group A and B were treated with oral administration of normal saline solution, 3 mL, weekly. The rats in the group C were treated daily with acupuncture at "Huantiao"(GB 30), "Housanli" (ST 36), "Yanglingquan"(GB 34) and "Weizhong"(BL 40) on the left hind legs; the rats in the group D were given orally Nilestriol solution (0.2 mg/mL) in a dose of 0.6 mL/100 g body weight, weekly. At the 7th, 14th, 21st and 28th days, some rats were sacrificed and their fractural callus and blood samples were taken for histological examinations and immunohistochemical examination of brain-derived neurotrophic factor (BDNF) and tyrosine kinase B (TrkB). RESULTS: HE stained callus slides observed by optical microscope showed that the process of fracture healing in the group A, C, D was faster than that in the group B. Positive immuno stalning of BDNF and TrkB could be seen in the all groups, mainly on the 7 and 14 days after the fracture. The expression levels from high to low in turn were group A, C, D and B. CONCLUSION: Expressions of BDNF and TrkB in callus of osteoporotic fracture were less than that of the normal fracture; acupuncture can elevate the expression levels and accelerate the process of fracture healing. PMID- 19565741 TI - [Study on the mechanism of electroacupuncture scalp point penetration therapy in action on apoptosis in the Parkinson's disease rat model]. AB - OBJECTIVE: To explore the mechanism of electroacupuncture scalp point penetration therapy in treatment of the Parkinson's disease (PD). METHODS: Forty Wistar rats were randomly divided into a normal group, a sham-operation group, a model group and an electroacupuncture (EA) group. 6-OHDA was injected into the left striatum to make lateralization PD rat model. Acupuncture at "Baihui" (GV 20)-through "Taiyang" (EX-HN 5), once each day, 6 days constituting one course. Immunohistochemical method was used to observe the facio-density and the integral optical density of brain-derived neurotrophic factor (BDNF) in the left substantia nigra, and TUNEL method was used to observe the apoptotic amount, and high performance liquid chromatography was used to observe DA contents of the left striatum in each group. RESULTS: As compared with the model group, in the acupuncture group, the facio-density and the integral optical density in the left substantia nigra increased significantly (P < 0.05), the amount of apoptosis decreased significantly (P < 0.05), and the content of striatum DA increased significantly (P < 0.05). CONCLUSION: EA scalp point-through-point therapy may enhance BDNF protein expression level in the substantia nigra to decrease the amount of apoptosis in the PD model rat. PMID- 19565742 TI - [Thinking about the conclusion of no difference between the acupuncture and sham acupuncture in the clinically therapeutic effects on migraine abroad]. AB - For the conclusion of "no difference between the acupuncture and sham-acupuncture in the clinically therapeutic effects on migraine abroad", by description of definition of sham-acupuncture (placebo acupuncture), the following principle needed by ideal placebo acupuncture, and definition, parts, functions and clinical effects of shallow needling, whether or not can shallow needling be used as the control method of sham-acupuncture in the clinical acupuncture study are explored, the results indicate that "the called sham-acupuncture (placebo acupuncture)" control method, i.e., "needling acupoints or non-acupoints on the superficial lay of skin with minimal stimulating amount" is very possibly activates the regulative action of the whole body through "superficially needling the skin part", to exert the therapeutic action. Therefore, this control method is not a proper control method of placebo acupuncture, and can not be used as the control method for sham-acupuncture (placebo acupuncture) in clinical acupuncture studies. Thus, the conclusion of "no difference between the acupuncture and sham acupuncture in the clinically therapeutic effects" made by this placebo acupuncture control method naturally may be overturned. PMID- 19565743 TI - [Study on key problems in the positive control design methods of acupuncture and moxibustion ]. AB - To probe into the design method of the clinical studies with positive control of acupuncture and moxibustion, the several key problems are analyzed, which include the definition of positive control, form of the control, statistical and design types, proper application of positive drugs, and properly use placebo control method to counteract weaknesses of studies, reasonably select observation indexes so as to show characteristics of Chinese medicine. The authors indicate that only fully thinking these key problems, can provide clinical evidence of acupuncture and moxibustion with high quality and convincingness. PMID- 19565744 TI - [Comment on the development of manipulation standardization of acupoint application technique]. AB - Acupuncture and moxibustion is a technique of TCM with the most tremendous influence in international, its standardization will be of important significance for promoting TCM widely spreading in international. Point application is one of important compositions of acupuncture and moxibustion techniques. Development of manipulation standardization of acupoint application technique is an important measure for increasing medical quality, ensuring medical safety and preventing medical risks, and this is of important significance for scientific studies and clinical popularization of point application technique. In the present paper, the difficult points, resolving way and standard technique characteristics, the attention problems and defects in usage in development course of manipulation standardization of acupoint application technique are discussed. PMID- 19565745 TI - [Observation on therapeutic effect of bee needle therapy on chronic lumbar muscle strain]. AB - OBJECTIVE: To explore a new method for treatment of chronic lumbar muscle strain. METHODS: One hundred and fifty cases were randomly divided into 2 groups, an observation group of 78 cases and a control group of 72 cases. The observation group was treated with bee-needle therapy, with Jiaji (EX-B 2) on the loin and Shenshu (BL 23), Zhishi (BL 47), Ciliao (BL 32), Weizhong (BL 40), Ashi points selected as main; and the control group was treated with routine acupuncture at the same points as those in the observation group, in combination with fire cupping or warm needle moxibustion. They were treated once daily, 10 sessions constituting one course. After 3 courses and a half year later, therapeutic effects were observed and followed up. RESULTS: In the observation group, 49 cases were cured, 27 improved, 2 ineffective, with a cured rate of 62.8%, and in the control group, the corresponding figures were 29, 40, 3 and 40.3%, with a significant difference between the two groups in the cured rate (P < 0.01). CONCLUSION: The therapeutic effect of the bee-needle therapy on chronic lumbar muscle strain is better than that of the routine acupuncture, which provides a better method for treatment of chronic lumbar muscle strain. PMID- 19565746 TI - [Development and usage of the along-meridian treatment instrument]. AB - The points stimulated by output pulse of the electroacupuncture instrument are fixed, and this stimulation model makes Deqi sense of the distal point unable to reach the affected parts, only limiting around the point for most patients, so the therapeutic effects are influenced to a certain extent. From design, usage and other aspects, it is described that joint the output conducting wires to the acupuncture needles from the distal point to the proximal point in turn. When the output ports of the electroacupuncture instrument export the pulse wave one port by one port, the sense of Deqi of the patient propagates towards the affected area one point by one point. Finally, the sense of Deqi of the patient propagates and reaches to the affected area. The along-meridian treatment instrument overcomes the shortcomings of the existing electroacupuncture instrument, and obviously increases therapeutic effects. PMID- 19565747 TI - [Safety problems and countermeasures of electroacupuncture instruments]. AB - According to the clinical experience and the active role of electroacupuncture instruments in clinical acupuncture and moxibustion, the potential security hazards of the available electroacupuncture instruments are pointed out. These hazards not only can cause injury to a varying degrees for the patient, but also lead to a poor therapeutic effect. The reasons of the security problems are mainly inaccurate regulation parameters, unreasonable displaying parameters, unscientific output of constant voltage, and unqualified protection measures for the electroacupuncture instrument. The countermeasures improving security and the measures increasing the intelligent level of the electroacupuncture instrument are put forward. PMID- 19565748 TI - [Survey of acupuncture and moxibustion for clinical treatment of renal diseases]. AB - In order to understand survey of medication combined with acupuncture and moxibustion for clinical treatment of renal diseases, clinical application and the mechanisms of acupuncture and moxibustion for treatment of renal diseases were summarized by electric retrieval of literature from 1982 to 2007. It is indicated that acupuncture and moxibustion can increase human immunity, reduce urinary protein, improve renal function, antagonize the side-effects of glucocorticoid hormones, etc. and medication combined with acup-moxibustion has the advantages of convenience, lower cost, safety, no adverse effects, etc. PMID- 19565749 TI - [Provenance difference in stable carbon isotope discrimination of Schima superba]. AB - The difference in leaf stable carbon isotope discrimination (delta) of 18 representative Schima superba provenances was investigated based on three provenance trails located at Jian' ou of Fujian Province and Chun' an and Qingyuan of Zhejiang Province, and the geographic variation pattern and the effects of trial site and provenances growth rate were studied. Significant differences in leaf delta were observed among the provenaces, and the differences in leaf delta value between the highest and lowest provenances reached 6.9%, 3.0%, and 3.7% in 3 experimental sites, respectively. A classic latitudinal clinical variation pattern was found, because there were significant correlations between the leaf delta value and the latitude of seed sources in all the 3 sites while no significant correlations were observed between the leaf delta value and the longitude of the seed sources. The southern provenances showed higher leaf delta value than the northern provenances, indicating that the water use efficiency (WUE) was lower in southern provenances. The leaf delta value of different provenances was demonstrated to be greatly affected by the environment of trial sites. The leaf delta value increased significantly with the improvement of site environment and with the increase of annual rainfall. Significant positive correlations were observed between the leaf delta value and the growth traits including tree height, DBH, total number of lateral branches, and length of the strongest lateral branch, which indicated that the provenances with higher growth rate and denser crown had larger leaf delta value. Two and four superior provenances with high growth rate and low leaf delta value (or high WUE) were selected for Jian' ou of Fujian and Chun' an of Zhejiang, respectively. PMID- 19565750 TI - [Effects of fertilization on nutrient concentrations of different root orders' fine roots in Larix kaempferi plantation]. AB - With the 16 years old Larix kaempferi plantation in eastern mountain area of Liaoning Province, China as test object, this paper studied the effects of fertilization on the nutrient concentrations of five root orders' fine roots. Under fertilization, less difference was observed in the total C concentration of the fine roots. Among the five orders' fine roots, the first order's had the lowest concentration of non-structural carbohydrate (TNC) and the highest ones of N and P, while the fifth order's was in adverse. The TNC concentration increased with increasing root order, while the N and P concentrations decreased correspondingly. Fertilization only had significant effects on the N and P concentrations of the first order's fine roots. The C/N/P ratio in different orders' fine roots had significant differences, being 423 : 16 : 1 and 726 : 16 : 1 in the first and the fifth order's fine roots, respectively. With the increase of root order, the proportion of C increased significantly, while that of N varied little. N fertilization didn't change the proportion of C, while P or P + N fertilization decreased the proportions of C and N in the first three orders' fine roots at 0-10 cm soil depth or in the first two orders' fine roots at 10-20 cm soil depth. PMID- 19565751 TI - [Effects of phosphorus stress on the growth and nitrogen and phosphorus absorption of different Formosan sweet gum provenances]. AB - Aiming at the ecological value of Formosan sweet gum (Liquidambar formosana) as a pioneer species and the status of red soil phosphorus (P) deficiency, a sand culture experiment of split design was conducted to study the responses of three leaf stage seedlings of seven Formosan sweet gum provenances from Yixing of Jiangsu, Jingxian of Anhui, Yongkang of Zhejiang, Nanchang of Jiangxi, Shaowu of Fujian, Yanping of Fujian, and Nandan of Guangxi to four levels of P (P0, P1/2, P1, P2). With increasing P stress, the biomass and the N and P absorption of test provenances decreased, whereas the utilization efficiency increased. In higher P treatments, the provenances from Nanchang and Yixing had higher biomass and higher N and P absorption but lower utilization efficiency, while the provenance from Nandan had lower N and P absorption but higher utilization efficiency. In lower P treatments, the biomass and the P absorption and utilization efficiency of the provenances from Nanchang and Nandan were all higher. All the results illustrated that the provenances with high biomass had high P absorption at high P level, and had both high P absorption and high utilization efficiency at low P level. The provenance from Nanchang could be considered to be an excellent P stress-resistant provenance, followed by that from Nandan. Phosphorus was not a limiting nutritional factor of Formosan sweet gum, biomass, leaf delta (N/P) ratio and P efficiency could be used as the indicators of P stress-tolerance of Formosan sweet gum provenances. PMID- 19565752 TI - [Responses of mulberry seedlings photosynthesis and antioxidant enzymes to chilling stress after low-temperature acclimation]. AB - Taking the seedlings of mulberry (Morus alba) variety 'Qiuyu' as test materials, this paper studied the changes of their leaf photosynthesis and antioxidant enzymes during low-temperature acclimation, chilling stress, and normal temperature recovery. The results showed that low-temperature acclimation at 12 degrees C for 3 days markedly increased the cold resistance of mulberry seedlings. After the acclimation, the leaf net photosynthetic rate (P(n)), stomatal conductance (G(s)), and maximum photochemical efficiency of photosystem II (F(v)/F(m)) under the chilling stress at 3 degrees C for 3 days increased obviously and recovered rapidly, compared with the control non-acclimation. During the period of the low-temperature acclimation and chilling stress, the leaf proline and soluble sugar contents increased. After the acclimation, the leaf malondialdehyde (MDA) content was obviously lower while the leaf ascorbate peroxidase (APX) activity was obviously higher than the control, illustrating that the increase of osmolytes content and APX activity played important roles in the cold resistance of mulberry seedlings. PMID- 19565753 TI - [Absorption and allocation characteristics of K+, Ca2+, Na+ and Cl- in different organs of Broussonetia papyrifera seedlings under NaCl stress]. AB - One-year-old Broussonetia papyrifera seedlings were subjected to 0.4, 1, 2, 3, and 4 g x kg(-1) of soil NaCl stress, and their biomass accumulation, leaf plasma membrane permeability, and the absorption, allocation and translocation of K+, Ca2+, Na+, and Cl-, as well as the symptoms of salt injury, were studied and investigated. The leaf plasma membrane permeability increased with the increase of soil NaCl concentration and of the duration of soil NaCl stress, and the seedling's root/shoot ratio also increased with increasing soil NaCl concentration. When the soil NaCl concentration exceeded 3 g x kg(-1), leaf plasma membrane permeability and seedling' s biomass accumulation were affected significantly. The Na+ and Cl- concentrations in different organs of seedlings increased with increasing soil NaCl concentration while the K+ and Ca2+ concentrations were in adverse, and the ion contents in leaves were always much higher than those in other organs, illustrating that soil NaCl stress affected the K+ and Ca2+ absorbing capability of roots, and inhibited the selective translocation of K+ and Ca2+ to aboveground parts. As a result, the K+ and Ca2+ concentrations in leaves and stems decreased. The study showed that B. papyrifera could effectively resist the injury of osmotic stress from soil salt via absorbing and accumulating Na+ and Cl-, but excessive accumulation of Na+ and Cl- could induce salt toxicity. As a non-halophyte species with relatively strong salt resistance, the aboveground parts of B. papyrifera did not have significant salt-exclusion effect. PMID- 19565754 TI - [Coarse woody debris loading capacity and its environmental gradient in Huzhong forest area of Great Xing' an Mountains]. AB - The coarse woody debris (CWD) loading capacity of main forest types and of different larch forest communities in Huzhong area of Great Xing'an Mountains was investigated, with its environmental gradient analyzed by Detrended Canonical Correspondence Analysis (DCCA). The CWD loading capacity varied significantly with forest type, being the highest (0.20 m3 x hm(-2)) in spruce forest (Picea koraiensis). Different larch forest (Larix gmelinii) communities had a CWD loading capacity from 0 to 0.28 m3 x hm(-2), with the highest in Pinus pumila Larix gmelinii (0.28 m3 x hm(-2)) and the lowest in Sphagnum magellanicum-Ledum palustre-Larix gmelinii (0), but the differences were not significant. The CWD loading capacity across the study area was complicated, which might result from the complex interplay of affecting factors. The main factors affecting the spatial pattern of the CWD loading capacity were topographic factors (elevation and slope position) and stand structure (age, canopy cover, and others), and the interaction between topographic factors and stand structure expressed the spatial gradient of CWD loading capacity in Huzhong area of Great Xing' an Mountains. PMID- 19565755 TI - [Growth analysis on modules of Cynodon dactylon clones in Yili River Valley Plain of Xinjiang]. AB - By the method of randomly digging up whole ramet tuft while maintaining natural integrity, large samples of Cynodon dactylon clones were collected from a grape orchard abandoned for 2 years without any management in the Yili River Valley Plain of Xinjiang, aimed to quantitatively analyze the growth patterns of their modules. The results showed that the average ramet number of test 30 clones reached 272.6 +/- 186. 6, among which, vegetative ramets occupied 82.3%, being 4.3 times higher than reproductive ones. The total biomass of the clones was 45.4 +/- 40.0 g, in which, rhizomes accounted for 54.4%, while the vegetative ramets, stolons, and reproductive ramets occupied 21.0%, 14.8%, and 9.4% of the total, respectively. The accumulative length of rhizomes and stolons reached 5.1 + 4.7 m and 3.3 +/- 3.4 m, while the bud number on stolons and rhizomes was 291.5 +/- 246.8 and 78.8 +/- 87.4, respectively. The bud number on stolons and rhizomes was positively correlated to the quantitative characters of vegetative ramets, reproductive ramets, stolons, and rhizomes (P < 0.01), indicating that in Yili River Valley Plain, C. dactylon clone could achieve and maintain its continuous renovation via rhizome buds. PMID- 19565756 TI - [Impact of climate warming on phenophase of Populus tomentosa in Inner Mongolia]. AB - Based on the 1982-2006 data of air temperature and of the phenophase of Populus tomentosa in spring and autumn from 17 phenological observation stations in Inner Mongolia, the change trend of the phenophase of P. tomentosa in spring and autumn was studied, and its relations to climate warming were analyzed. The models for predicting the date change of P. tomentosa first flowering and defoliating were constructed by stepwise regression method. It was shown that from 1982 to 2006, the air temperature in study area was increased, and the date of first flowering and defoliating of P. tomentosa was advanced and postponed, respectively. The first flowering of P. tomentosa was negatively related to the air temperature in spring and winter, and the main affecting factor was the air temperature from March to April; while the first defoliating of P. tomentosa was positively related to the air temperature in autumn, and the main affecting factor was the air temperature from August to October. According to the national assessment report on climate change, the air temperature in Inner Mongolia in spring and autumn would be raised by 1.5 degrees C-5.4 degrees C and 1.2 degrees C-4.4 degrees C, and the first flowering and defoliating of P. tomentosa would advance 3.9-17.8 days and postpone 3.0-12.4 days, respectively. PMID- 19565757 TI - [Effects of NaCl stress on Hippophae rhamnoides and Shepherdia argentea seedlings growth and photosynthetic characteristics]. AB - With two-year old seedlings of Hippophea rhamnoides and Shepherdia argentea as test materials, this paper studied their growth and photosynthetic characteristics under the stress of different concentration (0, 200, 400 and 600 mmol x L(-1)) NaCl. The results showed that the biomass and total leaf area per plant of H. rhamnoides and S. argentea seedlings decreased significantly with increasing NaCl concentration. Comparing with the control, the root/shoot ratio of H. rhamnoides and S. argentea seedlings under NaCl stress increased obviously, while the leaf mass per area (LMA) decreased slightly. When the NaCl concentration increased and the stress time prolonged, the net photosynthetic rate (P(n)), transpiration rate (T(r)), and stomatal conductance (G(s)) of H. rhamnoides and S. argentea seedlings declined markedly, the intercellular CO2 concentration (C(i)) increased after an initial decrease, whereas the water use efficiency (WUE) and stomatal limiting value (L(s)) decreased after an initial increase. The dynamic changes of G(s), C(i) and L(s) indicated that the decline of P(n) was mainly caused by the stomatal limitation in a short-term stress, and by non-stomatal limitation in a long-term stress. The poorer the salt tolerance of tree species and the higher the NaCl concentration, the earlier the transition from stomatal limitation to non-stomatal limitation would occur. As for H. rhamnoides, its morphological symptoms of salt injury appeared on the 10th day, and all of its seedlings were died on the 22th day under 600 mmol NaCl x L(-1) stress. In contrast, S. argentea could tolerate 600 mmol NaCl x L(-1) stress for above 30 days, illustrating that S. argentea, as an introduced tree species, had higher salt tolerance than H. rhamnoides, and could be planted widely in saline regions of China. PMID- 19565758 TI - [Recovery effect of tomato leaves photosynthesis and chloroplast ultrastructure after a short-term low nocturnal temperature]. AB - Tomato seedlings were treated with 15 degrees C (control), 12 degrees C, 9 degrees C, and 6 degrees C at night for seven days to study the changes of their leaf photosynthesis and chloroplast ultrastructure, aimed to make sure the recovery effect of the photosynthesis of tomato leaves after a short-term low nocturnal temperature. The results showed that short-term low nocturnal temperature was favorable to the photosynthesis of tomato leaves at recovery stage. The chlorophyll content could recover to the normal level as control under the treatments of 12 degrees C and 9 degrees C, but not under the treatment of 6 degrees C. The formation of stoma was not seriously inhibited by low nocturnal temperature. On the contrary, the stoma formation and its growth were stimulated at recovery stage. Under the treatment of 6 degrees C, chloroplast grana layers arranged loosely, and their number reduced. In the seven days of recovery, the lower the treatment temperature, the smaller and the fewer the starch grains were, and the more compact and tidy the chloroplast grana became. In addition, the transportation of photosynthesis products was promoted after short-term low nocturnal temperature. PMID- 19565759 TI - [Effects of shading on wheat grain starch quality and redistribution of pre anthesis stored nonstructural carbohydrates]. AB - This paper studied the effects of shading in the period from jointing to maturity on the grain yield, starch content, and starch paste traits of two winter wheat (Triticum aestivum L.) cultivars, shading-tolerant Yangmai 158 and shading intolerant Yangmai 11, and analyzed the relationships of the redistribution of total soluble sugars stored before anthesis in vegetative organs with the grain yield, starch content, and starch paste parameters of the cultivars. The results showed that shading decreased the redistribution of pre-anthesis stored total soluble sugars in vegetative organs to reproductive organ, resulting in the decrease of grain yield, and significantly decreased the amylopectin content but had no effects on the amylose content in grains, inducing a significant decrease in the ratio of amylopectin to amylase content in the grains. Shading also decreased the grain starch peak viscosity of the cultivars. Under shading, Yangmai 11 had a decrease of its grain starch through viscosity and an increase of starch pasting temperature, while Yangmai 158 had lesser responses in the two parameters. PMID- 19565760 TI - [Interactive effects of drought and salt stresses on winter wheat seedlings growth and physiological characteristics of stress-resistance]. AB - In a hydroponic culture, different concentrations of PEG-6000 (0, 8.3%, and 12.6%, W/V) and NaCl (0, 25, and 50 mmol x L(-1) were added to simulate different degrees of drought and salt stresses, aimed to study their interactive effects on the winter wheat (cv. Cang-6001) seedlings growth and physiological characteristics of stress-resistance. The results showed that under the conditions of adding 8.3% and 12.6% of PEG-6000, the addition of 25 mmol NaCl x L(-1) increased the dry matter accumulation and water content in plant, the contents of soluble sugar and soluble protein in leaf and the Na+ content in shoot and root, while decreased the MDA and proline contents in leaf and the K+ content in shoot and root, compared with no NaCl addition. Adding 12.6% of PEG 6000 and 50 mmol x L(-1) of NaCl more inhibited plant growth, compared with no NaCl added. It was suggested that under drought stress, applying definite amount of salt could alleviate the deleterious effects of drought stress on winter wheat seedlings growth. PMID- 19565761 TI - [Effects of returning straw to soil and different tillage methods on paddy field soil fertility and microbial population]. AB - A field experiment was conducted on a paddy field to study the effects of returning straw to soil and different tillage methods (no-tillage + returning straw, no-tillage, tillage + returning straw, and tillage) on the fertility level and microbial quantities of different soil layers. The results showed that in upper soil layer, the organic matter content in treatment 'no-tillage + returning straw' was 5.33, 2.79, and 5.37 g x kg(-1) higher than that in treatments 'no tillage', 'tillage + returning straw', and 'tillage', respectively, and the contents of total and available N, P and K in treatment 'no-tillage + returning straw' were also the highest, followed by in treatments 'no-tillage' and 'tillage + returning straw', and in treatment 'tillage'. In deeper soil layer, all the fertility indices were higher in treatment 'tillage + returning straw'. Treatments of 'returning straw to soil' had the highest quantities of soil microbes. The quantities of bacteria, fungi, and actinomycetes in upper soil layer were the highest in treatment 'no-tillage + returning straw', and thus, the cellulose decomposition intensity in this treatment at maturity period was 26.44%, 79.01%, and 98. 15% higher than that in treatments 'tillage + returning straw', 'no-tillage', and 'tillage', respectively. In deeper soil layer, the quantities of bacteria, fungi, and actinomycetes were the highest in treatment 'tillage + returning straw'. Treatment 'no-tillage + returning straw' had the features of high fertility and abundant microbes in surface soil layer. The quantities of soil bacteria and actinomycetes and the decomposition intensity of soil cellulose were significantly positively correlated with soil fertility level. PMID- 19565762 TI - [Effects of palygorskite application on volatilization and leaching losses of urea nitrogen in loess soil]. AB - By the methods of adsorption and soil column leaching, a laboratory simulation test was conducted to study the effects of palygorskite application on the volatilization and leaching losses of urea nitrogen in loess soil. Comparing with applying urea fertilizer alone, the addition of palygorskite could decrease the volatilization rate of soil urea N at peak time, with the ammonia volatilization loss decreased by 13.6%-15.0%. When the palygorskite application rate was 0.3 and 0.6 g x kg(-1), the leaching rate of soil NH4(+) -N and NO3(-) -N decreased, with the leaching loss of soil mineral N decreased by 13.7% and 13.6%, respectively. Applying 0.9 g x kg(-1) of palygorskite application increased the leaching rate of soil NH4(+) -N and NO3(-) -N, with the leaching loss of soil mineral N increased by 6.1%. Applying 0.3 g x kg(-1) of palygorskite increased soil NH4(+) N content by 0.20 mg x kg(-1), while applying 0.9 g x kg(-1) of palygorskite decreased soil NH4(+) -N content by 0.42 mg x kg(-1). Palygorskite application increased soil NO3(-) -N content by 1.24-2.52 mg x kg(-1). It was concluded that the application of palygorskite could decrease the volatilization rate of urea N, and applying appropriate amounts of palygorskite could decrease the leaching loss of soil mineral N and increase the contents of soil NH4(+) -N and NO3(-) -N. PMID- 19565763 TI - [Changes of soil biological characters beneath greenhouse cucumber under different cultivation systems]. AB - A 3-year pot experiment with 8-year continuously cropped greenhouse cucumber soil was conducted to study the effects of different cultivation systems on the soil quality, soil nematode quantity, and crop yields. The third year' s data were taken into analysis. Comparing with traditional cultivation system (two seasons planting cucumber with fallowing in summer) , planting leafy vegetable and garlic in summer increased the yields of the two crops and the soil microbial biomass carbon content, total microbial population, and sucrase activity, decreased the amounts of soil nematode and root-knot nematode, and maintained a relative high Shannon-Weaver index. Planting crown daisy, garlic, and spinach in summer increased soil sucrase activity by 8.9%, 89.5%, and 36.9%, and the planting of crown daisy and garlic also increased the Shannon-Weaver index by 7.7% and 9.4%, respectively. All the results suggested that catching crop and rotation had definite restoration effects on the soil quality under continuously cropping of cucumber, and the effects of planting crown daisy and garlic were more significant. PMID- 19565764 TI - [Antifeedant and antioviposition activities of Momordica charantia leaf ethanol extract against Liriomyza sativae]. AB - Liriomyza sativae Blanchard (Diptera: Agromyzidae) is one of the important pests harming a wide variety of vegetables and ornamental plants throughout the world. The leaf ethanol extract of Momordica charantia at the concentration of 2000-4000 microg x ml(-1) displayed significant antifeedant and antioviposition activities against L. sativae adults. For further purifying the extract, four solvents, i. e., cyclohexane, ethyl acetate, n-butanol and water, were used to extract the ethanol extract, and the antifeedant and antioviposition activities of the extracts against L. sativae adults were tested. The results showed that after treated with the extracts at the concentration of 1000 microg x ml(-1) for 2 days, the antifeedant index (AFI) of cyclohexane-, ethyl acetate-, n-butanol- and water extracts against L. sativae adults was 11.08%, 34.89%, 22.99% and 0, and the antioviposition index (AOI) was 0, 30.91%, 6.45% and 0, respectively. Ethyl acetate extract had the highest bioactivity. At the concentration of 4000 microg x ml(-1), the AFI and AOI of ethyl acetate extract were 70.95% and 69.49%, respectively. The ethyl acetate extract was then isolated by silica gel column chromatography, and three compounds, i.e., (19S, 23E)-5beta,19-epoxy-19-methoxy cucurbita-6,23-dien-3beta and 25-diol (compound 1), (19R, 23E)-5beta,19-epoxy-19 methoxy-cucurbita-6,23-dien-3beta and 25-diol (compound 2), and 3beta, 7beta,25 trihydroxycucurbita-5,23-dien-19-al-3-O-beta-D-glucopyranoside (compound 3), were obtained. These three compounds at concentration of 100-400 microg x ml(-1) all had inhibitory effects on the feeding and oviposition of L. sativae. At the concentration of 400 microg x ml(-1), the AFI and AOI were 66.89% , 53.53% and 78.02% , and 76.32%, 58.36% and 78.36% for compound 1, 2 and 3, respectively. PMID- 19565765 TI - [Risk assessment and control strategies of pests in Lycium barbarum fields under different managements]. AB - In the risk assessment of pests, both the community structure and the environmental factors should be considered at the same time, because of their mutual effects on the outbreak of disaster pests. This paper established a comprehensive assessment system, including 2 sub-systems, 5 respects, and 14 indices. In the meanwhile, risk assessment indices and experience formula were used to analyze the risk degree of pests in Lycium barbarum fields under different managements. It was found that using risk assessment indices and experience formula could obtain similar results. In abandoned field, Aceria palida, Aphis sp., and Paratrioza sinica were the frequent disaster pests, Lema decempunctata, Neoceratitis asiatica, Jaapiella sp., and Phthorimaea sp. were the incidental disaster pests, and Psylliodes obscurofaciata and Phthorimaea sp. were general pests. In organic field, the frequent disaster pests were the same species as those in abandoned field, while P. indicus, Jaapiella sp. and Phthorimaea sp. were the incidental disaster pests. In chemical control field, A. palida, Aphis sp., P. sinica, and P. indicus were the frequent disaster pests, while Jaapiella sp. and Phthorimaea sp. were the incidental disaster pests. Optimal 5 separations most fitted the division of pest sub-communities in L. barbarum fields, which were infancy period (from March 28 to April 15), outbreak I period (from April 15 to July 18), dormancy period (from July 18 to September 8), outbreak II period (from September 8 to October 15), and recession period (after October 15). The matrix of correlation coefficient showed that the dynamics of pests in L. barbarum fields under different managements were significantly correlated with each other, suggesting that the dynamics of pest populations was similar in different L. barbarum fields, which had two population establishment stages and one exponential growth stage in every year. The optimal controlling stages were from late infancy period to early and middle outbreak I periods, and from late dormancy period to early outbreak II period, which were very critical for pest control. PMID- 19565766 TI - [Quantitative analysis of insect pest and natural enemy communities in Red Fuji apple orchard]. AB - The insect pest and natural enemy communities in Red Fuji apple orchard in Mouping District of Yantai City were quantitatively analyzed by multivariate analysis. The temporal structure of the communities was grouped into five continuous stages by using optimal sorting method, and the community characteristics at each stage were described. The dominant pests and natural enemies were determined at different growth stages of apple trees through analyzing the sub-communities of the insect pests and the predatory and parasitic enemies by principal component analysis and factor analysis. Canonical correlation analysis showed that there were significant correlations between the dominant insect pests and the dominant natural enemies, especially between Lithocolletis ringoniella and its parasitoids, between Aphis citricola and its parasitoids, and between Tetranychus viennensis and its obligatory predatory enemies, Stethorus punctillum and Amblyseius orientalis. PMID- 19565767 TI - [UP-PCR diversity analysis of Fusarium population isolated from greenhouse melon soils]. AB - A total of 112 Fusarium isolates were obtained from 36 soil samples collected from the greenhouse melon fields of Liaoning Province, among which, 11 species were identified by traditional morphological classification and rDNA sequence analysis. Universally Primed PCR (UP-PCR) was conducted to analyze the 25 strains of test Fusarium isolates and 3 strains of positive control Fusarium isolates. The results indicated that a total of 73 bands appeared after amplification by using 6 primers, and 66 bands (90.4%) were polymorphic. The isolates were clustered into eight groups at the similarity of 0.736 by cluster analysis, among which, 14 isolates were clustered into one group. It was concluded that UP-PCR could present the genetic relationship and difference among Fusarium strains, being able to be used as an assistant method for Fusarium classification. PMID- 19565768 TI - [Soil bacterial community structure in primeval forest and degraded ecosystem in Karst region]. AB - By using PCR-RFLP, this paper studied the 16S rDNA gene diversity and phylogenesis of soil bacteria in primeval forest and degraded ecosystem in Karst region of Northwest Guangxi. More genotypes and higher diversity index were observed in the soil of primeval forest than in that of degraded ecosystem, and only two common genotypes were observed in the two soils. A clone from each genotype was randomly selected as representative for sequencing. The obtained 16S rDNA gene sequences had a similarity of 87%-100% with those in the GenBank (www. ncbi. nlm. nih. gov), and more than half of them had a similarity lower than 97%, being of new species. Based on phylogenetic analysis, the bacteria in the two soils were classified into 10 groups, with 5 groups in common. The dominant bacterial groups in the two soils differed obviously. In primeval forest soil, the dominant group was Proteobacteria, which had 39 genotypes, occupying 58.0% of all the clones; while in the soil of degraded ecosystem, the dominant groups were Acidobacteria and Proteobacteria, which had 19 and 15 genotypes, occupying 32.5% and 30.5% of all the clones, respectively. In the soil of degraded ecosystem, Proteobacteria group decreased while Acidobacteria group increased markedly, compared with those in primeval forest soil. Soil physical and chemical properties and environmental factors should be responsible for the difference of soil bacterial community between the two soils. PMID- 19565769 TI - [Prediction of winter wheat yield based on crop biomass estimation at regional scale]. AB - Based on the 2004 in situ data of crop yield, remote sensing inversed photosynthetically active radiation (PAR), fraction of photosynthetically active radiation (f(PAR)), climate, and soil moisture in 83 typical winter wheat sampling field of 45 counties in Shijiazhuang, Hengshui, and Xingtai of Hebei Province, a simplified model for calculating the light use efficiency (epsilon) of winter wheat in Huanghuaihai Plain was established. According to the crop accumulated biomass from March to May and corrected by harvest index, the quantitative relationship between crop biomass and crop yield for winter wheat was set up, and applied in the 235 counties in Huanghuaihai Plain region of Hebei Province and Shandong Province and validated by the official crop statistical data at county level in 2004. The results showed that the root mean square error (RMSE) of predicted winter wheat yield in study area was 238.5 kg x hm(-2), and the relative error was 4.28%, suggesting that it was feasible to predict winter wheat yield by crop biomass estimation based on remote sensing data. PMID- 19565770 TI - [Modeling of cotton boll maturation period and cottonseed biomass accumulation]. AB - Field experiments with different maturity cotton cultivars and sowing dates were conducted at different sites to quantitatively study the effects of cultivar characteristics, weather conditions (air temperature and solar radiation), and crop management variable (N application rate) on the cotton boll maturation period and cottonseed biomass accumulation. The cotton boll maturation period was simulated by using the scale of physiological development time. Based on the hypothesis of sink-determined, the cottonseed biomass accumulation model was then developed. The subtending leaf N concentration of cotton boll was simulated with a semi-empirical equation, and used as the direct indicator of the N nutrition effect on cottonseed growth and development. The model was tested by independent field data obtained in the Yellow River Valley (Xuzhou and Anyang) and the lower reaches of Yangtze River Valley (Huaian) in 2005. The simulated values of boll maturation period showed reasonable agreement with observed values, with a root mean square error (RMSE) of 2.25 days for cultivar DSC-1, of 2.61 days for cultivar KC-1, and of 2.75 days for cultivar AC-33B. The RMSE of cottonseed dry mass prediction was 9.5 mg x seed(-1) for KC-1 and 8.2 mg x seed(-1) for AC-33B, indicating that the model had a good prediction precision. PMID- 19565771 TI - [Effects of air temperature change on spring wheat growth at different altitudes in northwest arid area]. AB - Based on the 1981-2006 observation data from agricultural meteorological stations at Minle (high altitude) and Zhangye (low altitude) in northwest arid area, the effects of air temperature change at the two altitudes on the growth and yield of spring wheat were studied. It was shown that during study period, the air temperature at the two altitudes had an increasing trend, and the increment was greater at high altitude than at low altitude. At high altitude, the growth duration of spring wheat shortened but the grain yield increased; while at low altitude, the growth duration shortened and the yield decreased. When the mean daily air temperature during spring wheat growth period increased by 1 degrees C, the growth duration shortened by 8.3 days at high altitude and by 3.8 days at low altitude. The growth duration and grain yield of spring wheat at high altitude had a slight increase when the maximum air temperature during growth period was below 30.4 degrees C, but decreased when the maximum air temperature was above 30.4 degrees C. PMID- 19565772 TI - [Assessment of farmland soil quality under different utilization intensity in arid area]. AB - Based on the 2005-2007 experimental data in Cele oasis in the southern margin of Tarim Basin of Xinjiang, the soil quality of four typical types of farmland with different utilization intensity, i.e., farmland with high input, farmland with normal input, newly reclaimed farmland, and farmland in oasis' interior, was analyzed and assessed by using sustainable yield index, soil improvement index, and soil quality synthesis index. Among the farmlands, there were significant differences in the contents of soil organic matter, available nitrogen, and available phosphorus. Newly reclaimed farmland had the lowest level of soil quality, while the farmland in oasis' interior had relatively higher soil quality. This study could help the reasonable exploitation and utilization of farmlands in Cele oasis, and the protection of local farmland eco-environment. PMID- 19565773 TI - [Spatiotemporal dynamics and landscape pattern of alien species Spartina alterniflora in Yancheng coastal wetlands of Jiangsu Province, China]. AB - Based on the 1992-2007 remote sensing images and field survey data, and by using the landscape ecology theories and the method of conversion matrix, the spatiotemporal dynamics and landscape pattern of Spartina alterniflora in Yancheng coastal wetlands were analyzed by GIS. The results showed that the total area of S. alterniflora along the whole coastal beaches of Yancheng increased from 3561 hm2 in 1992 to 14491 hm2 in 2007, with a growth rate of 306.94%. In 1992-2007, the total area of S. alterniflora conversion-in and conversion-out was 26291 hm2 and 15361 hm2, respectively, and the S. alterniflora community in the core area of Yancheng National Nature Reserve expanded from 597 hm2 to 2814 hm2, with an annual growth rate of 24.74%. The S. alterniflora community transformed from scattered patches in estuarine regions into continuous belt pattern mainly distributed in the periphery of coastal wetlands from Sheyang River to Liangduo River. Its centroid moved to the southeast, with a distance of 2.92 km. The average area of patches increased from 1992 to 2002, and then decreased from 2002 to 2007. The largest patch index and area-weighted contiguity index increased year by year, and the shape of patches tended to be more regular. PMID- 19565775 TI - [Population dynamics of Alexandrium tamarense and its relations with environmental factors in Nanji Islands sea area of Zhejiang Province]. AB - Based on the investigation in Nanji Islands sea area of Zhejiang Province from April 2006 to March 2007, the dynamics of Alexandrium tamarense and its relations with environmental factors were studied. In the study sea area, A. tamarense cells could only be found in spring (from April to June), and the cell abundance was of single peak curve, with the highest (12250 cells x L(-1)) in surface water on 8th of May. Higher cell abundance of A. tamarense was found at the water temperature of 18.5 degrees C - 19.5 degrees C, salinity of 29.5 per thousand - 31.0 per thousand, and lower PO4-P concentration, but no correlation was observed between A. tamarense cell abundance and water dissolved inorganic N concentration. Linear stepwise regression revealed that there was a significant positive correlation between the cell abundance of A. tamarense and the ratio of water dissolved inorganic N to PO4-P. PMID- 19565774 TI - [Neighborhood relationships of land use spatial pattern in Qixia City]. AB - By using the modules of neighborhood statistics and spatial analysis in ArcGIS software, an analysis was conducted with neighborhood factors on the spatiotemporal variation trend of the neighborhood relationships among main land use types in Qixia City of Shandong Province from 1987 to 2003. The results indicated that the neighborhood relationships between different land use types increased with increasing neighborhood distance, while those among the same land use types manifested congregation, which decreased with increasing neighborhood distance. From 1987 to 2003, the neighborhood relationships between construction land and orchard land, and between woodland and orchard land presented decreasing trend, while those of construction land with woodland and grassland were in adverse. Some measures such as controlling construction land area, protecting cultivated land, and decreasing the disturbances of human activities on woodland and grassland should be carried out to realize the harmonious development of economic and ecological benefits in Qixia City. PMID- 19565776 TI - [Seasonal dynamics of macrozoobenthos community structure in Xiangxi River]. AB - To study the seasonal dynamics of macrozoobenthos community in Xiangxi River, the individuals of macrozoobenthos in the River were quantitatively monitored. A total of 197 taxa belonging to 6 class and 68 families were collected, among which, Baetis spp., Epeorus spp., and Nemoura spp. were the dominant groups, and their compositions varied with seasons. The macrozoobenthos community was most complicated in winter, followed by in spring and autumn, and the simplest in summer. Gather-collector was the main component of functional feeding groups in Xiangxi River system, followed by scraper, predator, and filter-collector, while shredder only accounted for a small part. Canonical correspondence analysis showed that the factors affecting the macrozoobenthos community were dissimilar among seasons. In summer and autumn, nutrients had greater effects; and in all seasons, water depth had definite effects. PMID- 19565777 TI - [Structural characteristics of chironomid community and their indicative significance in bioassessment of water quality in Mingzhu Lake of Chongming Island, Shanghai]. AB - The structural characteristics of chironomid (Diptera: Chironomidae) community in Mingzhu Lake of Chongming Island, Shanghai, China were studied from July 2006 to April 2008. A total of eleven chironomid species belonging to 10 genera and 3 subfamilies were collected, among which, Propsilocerus akamusi and Microchironomus sp. were predominant. Based on clustering analysis (CA) and importance value (IV), it was found that the dominant species shifted seasonally, with Microchironomus sp. in spring and summer, Tanypus chinensis in autumn, and P. akamusi in winter. There was a significant seasonal difference in the Shannon Weaver diversity index (H') of the chironomid community, with the lowest in summer and higher in the other three seasons. The H' was significantly correlated with species richness (S) and evenness (J), and more strongly governed by J. The average density and biomass of the chironomid community were the highest in winter, with a minimum density in autumn and a lower biomass in the other seasons. Among the three dominant species, P. akamusi and Microchironomus sp. had more obvious seasonal fluctuations in their population dynamics than T. chinensis. Based on the bioassessment methods of water quality, e.g., H' and Hilsenhoff biotic index (BI), the Mingzhu Lake was considered to be moderately organically polluted and eutrophicated. PMID- 19565778 TI - [Effects of Pb and Ni stress on antioxidant enzyme system of Thuidium cymbifolium]. AB - This paper studied the accumulation and scavenging of active oxygen radicals, injury of photosynthetic system and membrane system, and changes of antioxidant enzyme system in Thuidium cymbifolium cells under single and combined stress of Pb and Ni. Under low concentration of Pb and Ni (Pb < 0.1 mmol x L(-1), Ni < 0.01 mmol x L(-1)), the chlorophyll content of T. cymbifolium increased; while under high concentration of Pb and Ni (Pb > 0.1 mmol x L(-1), Ni > 0.01 mmol x L(-1)), it was in adverse. There was a dose-dependent accumulation of reactive oxide species (ROS) and malondialdehyde (MDA) under combined Pb and Ni stress. The superoxide dismutase (SOD) and catalase (CAT) activities decreased while the peroxidase (POD) activity increased with increasing Pb and Ni concentration, indicating the important role of POD in eliminating ROS under Pb and Ni stress. The dose-dependent change of MDA content and CAT activity under Pb and Ni stress suggested that T. cymbifolium could be used as a biomarker in pollution monitoring of these two heavy metals. PMID- 19565779 TI - [Effects of exogenous tetracycline on rape soil enzyme activity and rape quality]. AB - This paper studied the effects of applying different concentration (0, 0.30, 0.60, and 0.90 mg x kg(-1)) tetracycline on the soil enzyme activities and rape quality. The results showed that soil urease activity after applied 0.30, 0.60, and 0.90 mg x kg(-1) of tetracycline and soil suerase activity after applied 0.90 mg x kg(-1) of tetracycline were inhibited in the whole cultivating period, while the soil sucrase activity after applied 0.30 and 0.60 mg x kg(-1) of tetracycline showed a trend of activation-inhibition-activation. Soil proteinase activity showed a trend of inhibition-activation, and the extent and duration of the inhibition and activation had significant positive correlations with tetracycline concentration (r = 0.950 * *). Soil catalase activity showed activation first and turned to irregular then. The action duration of tetracycline on the activities of soil urease, catalase, sucrase and proteinase were 7 weeks, 6-8 weeks, 7 weeks, and 6-7 weeks, respectively. At harvest time, the soluble sugar contents in rape leaves after applying 0.30, 0.60, and 0.90 mg x kg(-1) of tetracycline decreased dramatically by 91.99%, 87.92%, 90.12%, while the soluble protein content increased by 26.47%, 28.13%, and 23.22%, respectively, compared to the control. PMID- 19565780 TI - [Effects of enrofloxacin on DNA sequence diversity of soil cultural bacterial community]. AB - In order to understand the effects of remained enrofloxacin (ENR) in environment on the diversity of soil microbial communities, amplified ribosomal DNA restriction analysis (ARDRA) approach and genomic fingerprinting technique enterobacterial repetitive intergenic consensus sequence-polymerase chain reaction (ERIC-PCR) were used to analyze the molecular diversity of 16S rDNA from soil bacteria after ENR addition. The results showed that after the ENR addition for 35 days, the total count of soil bacteria was less than that of CK, and decreased with increasing ENR concentration. The ARDRA divided the separated soil bacteria into different operational taxonomic units (OTU) groups, and the count of group I to group VI was 15, 13, 10, 8, 6, and 6, respectively. Genomic fingerprinting analysis indicated that the Shannon-Wiener index of group I to group VI was 2.78, 2.14, 1.78, 1.11, 0.69 and 0.31, respectively, and the Margalef index, Simpson index, and Pielou index of soil microbial community in CK were higher than that in the soils in which ENR was added. PMID- 19565781 TI - [Relationships between economic growth and industrial pollutant discharge of Suzhou: how about the EKC?]. AB - Based on the study of the relationships between economic growth and industrial pollutant discharge of Suzhou City in 1991-2005, the environmental Kuznets curve (EKC) of the City was estimated by several kinds of function, with the cause analyzed. The results showed that the EKC of Suzhou was not a typical one. The industrial pollution of the City increased dramatically by scale effect of economic growth and the clear environmental benefit of industrial structure adjustment was not received obviously, while generalized discharge reduction technologies showed great results. In general terms, the pollution increase was comparatively slower than the GDP growth. As a whole, the environmental policy of Suzhou was in effect. However, the relationships between economic growth and industrial pollutant discharge had not been a benign development yet. In order to achieve win-win situation of economic growth and pollution reduction, Suzhou needed to optimize its industry structure, push the energy conservation and discharge reduction policy deeply, increase environmental investment, and enforce the strictest environmental and industrial policy. In doing these, Suzhou could turn the traditional omega-shaped EKC into delta-shaped EKC. PMID- 19565782 TI - [Dynamics of suspended solids in Xiangxi Bay of Three Gorges Reservoir during spring algal bloom period]. AB - The dynamics of total suspended solids (TSS) in Xiangxi Bay of Three Gorges Reservoir (TGR) during the period of spring algal bloom were weekly investigated. The results showed that the TSS in the Bay ranged from 0.66 to 134.92 mg x L(-1), with a median of 6.80 mg x L(-1), and tended to be increased from headwater to down-lake zones. The spatial pattern of non-volatile suspended solids (NVSS) was similar to that of TSS, while volatile suspended solids (VSS) showed obviously different pattern from TSS and followed the spatial pattern of chlorophyll a. Regression analysis indicated that in the mid-zone of the Bay, chlorophyll a was significantly related to TSS and VSS, and explained 66.7%-96.7% and 58.9%-85.5% of the variance in TSS and VSS, respectively. In the headwater and down-lake zones, there were no significant relationships between chlorophyll a and TSS (including NVSS and VSS). The TSS in mid-lake had more close relationship with VSS than with NVSS, while that in headwater and down-lake had more close relationship with NVSS than with VSS, implying that the mid-lake of Xiangxi Bay was a more autochthonous system than the headwater and down-lake where allochthonous influences were the more determinant factors. PMID- 19565783 TI - [Effects of drip irrigation under mulching on cotton root and shoot biomass and yield]. AB - By using bidirectional sampling method with soil drill, the effects of different amounts of drip irrigation (2618, 2947, 3600 and 4265 m3 x hm(-2)) under mulching on the root distribution, aboveground growth, and yield of cotton was studied in field. The results indicated that irrigation amount affected the root and shoot growth significantly. In all irrigation treatments, cotton root was mainly distributed in mulched area, occupying 60.65%-73.45% of total root biomass, while only 39.35%-26.55% was distributed in bare area. Water stress increased rooting depth, root biomass, and the extent of lateral rooting. Significant differences were observed in the biological characteristics and the biomass accumulation and allocation of cotton plant among different irrigation treatments. Over-irrigation (4265 m3 x hm(-2)) increased plant height, width of inverse fourth leaf, and amounts of branch and bud, and thus, accelerated biomass accumulation rate. Over irrigation also increased the root/shoot ratio and the proportion of biomass allocated to vegetative organs, but increased the fruit abscission rate and therefore reduced the economic yield. It was suggested that both excessive soil moisture content and water stress could affect the biomass accumulation and allocation in different cotton organs and at various life stages. Under the conditions of our experiment, 3600 m3 x hm(-2) was the optimal irrigation amount. PMID- 19565784 TI - [Environmental behavior and effect of biomass-derived black carbon in soil: a review]. AB - Biomass-derived black carbon, also named biochar, has the characteristics of high stability against decay and high capability of adsorption, and can affect the environment through its interactions with climate and geology, playing a significant role in global climate change, carbon biogeochemical cycle, and environmental system. In recent years, more and more researchers in the fields of atmospheric sciences, geology, and environmental science focused on the environmental behavior and effect of biochar. As one possible source of the components with high aromatic structure in soil humus, biochar is of great importance in increasing soil carbon storage and improving soil fertility, and in maintaining the balance of soil ecosystem. This paper offered the latest information regarding the characteristics and biotic and abiotic oxidation mechanisms of biochar, its effects on global climate change, and the environmental effect of biochar in soil. Research prospects were briefly discussed on the environmental behavior and effect of biochar in soil ecosystem. PMID- 19565785 TI - [Research progress on index system of regional ecological risk assessment]. AB - Regional ecological risk assessment (RERA) covers the assessments of multiple risk sources, receptors, and endpoints, while the selection of assessment indices is quite complicated, being a hotspot in regional environment management research. Domestic and international researches on RERA revealed that three processes in RERA are of vital, i.e., risk probability assessment measured by risk probability index, status and value assessment of ecosystem at regional scale indicated by ecological index, and vulnerability assessment of each ecosystem in a region under risk measured by vulnerability index. The main problems in the establishment of RERA index system are the strong subjectivity and poor comparability, and thus, the index system should be set up in the three key processes under the principles of objectivity, integration, hierarchy, and comparability. Due to the fact that the status and value assessment of ecosystem is most complicated, the index system should be formulated by compulsory and optional components to increase the comparability of RERA results between regions. PMID- 19565786 TI - [Present status, mechanisms, and control techniques of nitrogen and phosphorus non-point source pollution from vegetable fields]. AB - Chemical fertilizers are often excessively applied on vegetable fields to pursue higher yields. In some areas, the fertilization rates are several times of those needed by vegetables. Nitrogen and phosphorous are obviously accumulated in the vegetable soils, resulting in the malnutrition, excessive nitrate, and poor quality of vegetables. Furthermore, a series of environmental problems, e.g., deterioration of vegetable soil physical and chemical properties, nitrate pollution of groundwater, and eutrophication of surface water, are produced. This paper reviewed the present status of nitrogen and phosphorous non-point source pollution from vegetable soils (accumulation characteristics of nitrogen and phosphorous and their pollution risks to water environment and vegetables), pollution mechanisms (sources, transformation, and losses of nitrogen and phosphorous), and control techniques (fertilization, chemical addition, nitrogen catch crop cultivation, optimal planting system, spatial matching of source and sink landscapes, and grass buffer strip technology), aimed to supply references for the further study on the nitrogen and phosphorous non-point source pollution from vegetable fields. PMID- 19565787 TI - [Effects of UV-radiation on biological characteristics of different body-color biotypes of Sitobion avenae (Fab.)]. AB - UV-radiation exerts strong selection stress on the evolution of aphid populations, and thus, leads to their genetic differentiation. However, the effects of UV-radiation on different body-color biotypes of aphids are still ambiguous. In this study, new-born nymphae of red and green biotypes of Sitobion avenae were placed on two wheat varieties (Xiaoyan-22 and Astron), bred in an artificial bioclimatic chamber under strict controlled conditions (at 15 degrees C, 20 degrees C, and 25 degrees C, and treated with 30 W lamp of UV-B for 30 min per day for 5 days), and their development duration, mass, and mean relative growth rate were measured. The results showed that at lower temperature, UV radiation delayed the growth of green biotype aphid on Xiaoyan-22 and Astron significantly; while at higher temperature, UV-radiation significantly delayed the growth of red biotype aphid on Xiaoyan-22, but had lesser effects on the growth of the two biotypes on Astron, illustrating that different biotypes of aphids had different responses to UV-radiation, and the responses were correlated to temperature and wheat varieties. PMID- 19565788 TI - [The study of EEG Higher Order Spectral Analysis technology]. AB - The basic theory of Higher Order Spectral Analysis and the most generally used Bispectrum are introduced in the paper. By certain experiments of EEG signal acquisition and bispectrum analysis, it is showed that the Higher Order Spectrum has an advantage over power spectrum, which is based on Second Order Statistics, in processing non-linear signal and restraining Gauss noise signal. PMID- 19565789 TI - [A method to enhance user experience of EMR based on mining association rules of incremental updating data]. AB - The user experience (EX) of current Electronic Medical Record systems (EMR) is needed to improve. This paper proposed a new method to enhance EX of EMR. Firstly, system template and text characterization are used to make the EMR data structured. Then, the structured date are mined based on mining the association rules of incremental updating data to find the association of the elements of template of EMR and the values of elements. Finally, with the help of mined results, the users of EMR are able to input data effectively and quickly. PMID- 19565790 TI - [An adaptive scaling hybrid algorithm for reduction of CT artifacts caused by metal objects]. AB - A new adaptively hybrid filtering algorithm is proposed to reduce the artifacts caused by metal in CT image. Firstly, the method is used to preprocess the projection data of metal region and is reconstruct by filtered back projection (FBP) method. Then the expectation maximization algorithm (EM) is performed on the iterative original metal project data. Finally, a compensating procedure is applied to the reconstructed metal region. The simulation result has demonstrated that the proposed algorithm can remove the metal artifacts and keep the structure information of metal object effectively. It ensures that the tissues around the metal will not be distorted. The method is also computational efficient and effective for the CT images which contains several metal objects. PMID- 19565791 TI - [A novel respiratory detecting system based on bio-impedance]. AB - This paper introduces the design and implementation of a novel respiratory detecting system based on bio-impedance method. By increasing electrodes in space, the system make multi-channel respiratory signals be superpositioned and filtered (SNR); Traditional filter methods by both hardware and software are also used to further increase anti-interference ability. A low consumption and portable instrument is designed based on MSP430 Micro Controller Unit (MCU), The experiment shows a better performance in the reduction of interference noises of heartbeat and blood flow especially the motion artifact. Also the system works stably. PMID- 19565792 TI - [A study on the individual adaptability of electronic sphygmomanometer]. AB - This paper analyzed the limitation of electronic sphygmomanometer based on oscillometry, and according to the characteristics of pulse signal, the author proposed a new method of the characteristic parameter detection based on wavelet transform, and improved its recognition ability on fixed-scale, via resampling rate according to the heartbeat. And the prototype test has been proved that this method is more adaptability for individuals and stability for operation. PMID- 19565793 TI - [Fall detection and reliable device design]. AB - Utilizing the three dimensional accelerations measurement and the four-quadrant inclination switch which is developed to send an orientation warning, the fall detection method combining the impact detection and orientation prewarming is presented. And the accuracy of the fall detection is effectively improved. PMID- 19565795 TI - [Current research and development trend of implantable neuro-stimulator]. AB - This paper introduces the current application of various kinds of implantable neuro-stimulator at home and abroad, and also analyzes its development trend. PMID- 19565796 TI - [Study of axial flow pumps for ventricular assist]. AB - After introducing and comparing the most typical of axial-flow blood pumps in the world, this paper discusses the future development of axial blood flow pumps. PMID- 19565794 TI - [Deeply cooled far-infrared thermal imaging strategy for early tumor diagnostics]. AB - This paper is dedicated to evaluate the thermal behavior of skin surface embedded with tumor tissue through construction of three-dimensional heat transfer model of the human body. It was found that the far-infrared imaging equipment could not yet get the accurate results for diagnosis of tumors developed in early stage or located deeply in the human body, because of limited resolution and accuracy in the current system. Conceptual experiments with a thermal imaging system under various cooling levels were performed to confirm this issue. A dual cooling cavity was proposed to realize ultra-low-temperature so as to improve the cooling of the current infrared equipment and thereby to enhance its image precision and accuracy. This study is expected to be of significant reference value for realizing an early diagnosis of cancers through medical image. PMID- 19565797 TI - [Formation and suppression technique for motion and flow artifacts on magnetic resonance imaging]. AB - During magnetic resonance imaging, motion and flow artifacts often severely decrease image quality. The mechanisms of these kinds of artifacts are very complex. This article reviewed the pre-handling techniques of motion and flow artifacts, such as changing magnetic resonance imaging parameters, using spatial pre-saturation pulse sequences, gating and triggering techniques of suppressing respiratory motion artifact, as well as the progress of magnetic resonance pulse sequences in suppressing motion and flow artifacts, The paper briefly introduced post-processing techniques of suppressing motion and flow artifacts. PMID- 19565798 TI - [Electronic regulation mode on implantable medical device]. AB - OBJECTIVE: To discuss the methods of implantable medical device supervision reform. METHODS: Methods of literature review, comparative study, the research of situation, developmental study and case study were adopted. RESULTS: It is the untrue and asymmetric information of implantable medical device that results in improper behavior in the implantable medical device industry. CONCLUSIONS: A implantable medical device electronic monitoring system, namely establishing the independent but interrelated real-time dynamic updating of the databases, which would guarantee the information of implantable medical device is open and transparent, eliminate false food information transmission, vindicate the good order of domestic implantable medical device industry and protect the vital interests of the public. PMID- 19565799 TI - [The application of total quality management (TQM) in quality management of radiation therapy]. AB - The strategies and methods of the total quality management (TQM) need to applied in quality management of radiation therapy. We should improve the level of quality control and quality assurance in radiation therapy. By establishing quality control system in radiation therapy, standardization of radiation therapy workflow, strengthening quality control of devices and physical technique and paying attention to safety protection and staff training. PMID- 19565801 TI - [Designing the architecture of hospital equipment information system]. AB - This paper discussed the basic elements and the key technology of developing hospital equipment information system (HEIS), proposed the compatible fields' standard, and designed the system architecture and the core database structure. Following the method, we can establish robust HEIS, which can satisfy hospitals' equipment managing regulations, and accord with the work flow. The system has higher accuracy, efficiency and capability to counter risk. PMID- 19565800 TI - [Evidence-based management of medical disposable materials]. AB - Evidence-based management of medical disposable materials pays attention to collect evidence comprehensively and systematically, accumulate and create evidence through its own work and also evaluate evidence strictly. This can be used as a function to guide out job. Medical disposable materials evidence system contains product register qualification, product quality certification, supplier's behavior, internal and external communication evidence. Managers can find different ways in creating and using evidence referring to specific inside and outside condition. Evidence-based management can help accelerating the development of management of medical disposable materials from traditional experience pattern to a systematic and scientific pattern. It also has the very important meaning to improve medical quality, control the unreasonable growth of medical expense and make purchase and supply chain be more efficient. PMID- 19565802 TI - [Design and research of hospital medical supplies management information system]. AB - This paper introduces an advanced means to confirm management objective, analyze management need, reduce purchase and operating cost, optimize the flow management and establish a medical supplies management information system in purchasing, using, maintaining and disposing step. The system has advantage in realizing efficiency analyze, improving service and quality, guaranteeing safely use of medical supplies. PMID- 19565803 TI - [Management of large medical equipment of hospital]. AB - Medical equipment from the characteristics and the current situation, and explore the new situation to adapt to the equipment management model. A hospital medical equipment, large-scale integration of management concepts, multilevel personnel structure are satisfactorily managed a necessary condition for large-scale equipment. PMID- 19565804 TI - [Experience of preventive maintenance of medical equipment]. AB - Preventive maintenance of medical equipment can be helpful in improving the efficiency of medical devices, establishing a standardized management and plan, a classification of maintenance of medical devices, inspection tour of devices and operating condition of dynamic monitoring devices. It has benefit in prolonging the useful life of medical devices, reducing maintenance cost and improving management and service quality. PMID- 19565805 TI - [Maintenance and complications of hemodialysis equipment]. AB - Through the hemodialysis machine and water treatment systems maintenance, the result of complications caused by dialysis can be eliminated. It also of extended the life of hemodialysis machines and improve the quality of dialysis. PMID- 19565806 TI - [Tuberculosis hospitals in Russia: requirements, prospects for development]. AB - The results of treatment in patients with tuberculosis are low. Within the first 3-4 months of therapy, lung caverns are closed in 12-26% of patients. The length of tuberculosis inpatient treatment is 79 days, as provided by the state guarantees of rendering free treatment, and the proposals to reduce the number of beds are not substantiated. It is necessary to optimally restructure a bedspace and to bring it into proper sanitary position. PMID- 19565807 TI - [Role of health care facilities in verifying the diagnosis of respiratory tuberculosis by laboratory methods]. AB - Reasons for laboratory misdiagnosis that is observed in general health care network (GHCN) and penitentiaries were studied. The clinical efficiency of the diagnosis of tuberculosis in the GHCN by the Ziehl-Neelsen method is largely determined by three major factors, each of which is fundamentally important in assessing the result. The first factor is the adequacy of making up a cohort of patients having the symptoms of productive cough in the course of 2-3 weeks or more who are to be examined for tuberculosis by laboratory studies. The second is the quality of a diagnostic material. The third is the quality of laboratory specialists' work. The paper shows it necessary to implement programs for the internal control and external assessment of the quality of laboratory studies and to optimize (centralize) the activities of laboratories under the conditions of a regional network. PMID- 19565809 TI - [Roncoleukin in enhancing the efficiency of complex therapy for infiltrative pulmonary tuberculosis in adolescents]. AB - Seventy-eight patients, aged 15.69 +/- 0.15 years, who had infiltrative pulmonary tuberculosis, were examined. Roncoleukin was given intravenously in 20 patients of them (Subgroup 1a), in an indirect lymphotropic fashion in 30 patients (Subgroup 1b); 28 adolescents were treated with the conventional treatments (Group 2). A decay phase was recorded in 33 (66%) from Group 1 and in 16 (57.1%) from Group 2. On admission, all 78 (100%) patients had immunological disorders that required immunomodulation. Roncoleukin caused positive clinical and X-ray changes that were more pronounced in Subgroup 1b: bacterial excretion cessation was recorded in 100% of patients in Subgroup 1a (p < 0.05); in 94.4% in Subgroup 1b (p < 0.05), and in 68.7% in Group 2; decay cavities closed in 69.2, 60, and 37.5% in Subgroups 1a and 1b and Group 2. At the same time, the duration of a procedure considerably reduced, which positively affected adolescents' compliance. PMID- 19565808 TI - [Retrosternal lymphotropic chemotherapy and pneumoperitoneum in the treatment of drug-resistant destructive pulmonary tuberculosis]. AB - The efficiency of combined treatment with retrosternal lymphotropic administration of drugs and pneumoperitoneum was comparatively analyzed in 148 new cases of destructive pulmonary tuberculosis. Both methods were applied during combined chemotherapy in a study group of 48 patients. In Group 2 (n = 54), lymphotropic procedure was employed alone during standard chemotherapy; in Group 3 (n = 46), only pneumoperitoneum was used in combination of chemotherapy. In 40 patients, pulmonary tuberculosis was acutely progressive. Drug resistance was identified in 81.7%, including multidrug resistance in 27.7%. With the combined use of retrosternal lymphotropic therapy and pneumoperitoneum, bacterial excretion ceased in 96.2% of new cases of drug-resistant tuberculosis; decay cavity resolved in 80.8%. The proposed therapeutic technology used in patients with persistent bacterial excretion and decay cavities after ineffective 4-9 month treatment could achieve abacillation in 90.9% of patients and resolve decay cavities in 77.3%. Retrosternal drug injection reduces a risk for complications due to bronchial tuberculosis involvement when pneumoperitoneum is applied. PMID- 19565810 TI - [Assessment of a risk for postoperative infectious complications in patients with fibrocarvous pulmonary tuberculosis]. AB - The paper analyzes whether it is possible to predict a risk for postoperative empyema, by studying a totality of characteristics of a patient, a pathological process, and the properties of MBT in 46 patients with progressive fibrocavernous tuberculosis. It also shows it actual to accomplish this task with a prediction accuracy of 89.5-100%, by simultaneously taking into account both the bacteriological properties of MBT (the magnitude of Mycobacterium tuberculosis excretion and viability) and different combinations of serum biological parameters that reflect the activity of an inflammatory process. PMID- 19565811 TI - [Perioperative radiodiagnosis in children and adolescents with pulmonary tuberculosis]. AB - Routine X-ray examination indicated that the rate of cavernous and fibrocavernous tuberculosis was 21.4 and 25.9%, respectively. Computed tomography (CT) showed that the ratio of patients with cavernous tuberculosis to those with fibrocavernous one changed and was equal to 14.3 and 33.0%, respectively. The number of patients diagnosed as having fibrocavernous tuberculosis increased. According to the traditional studies, the number of patients with one lobe being affected turned out to be higher. Accordingly, that of patients with involvement of the whole lung or two lungs proved to be underestimated at cavitary analogue X ray study (AXS) or linear tomography (LT). Preoperative X-ray CT detected a larger number of caverns and tuberculomas. While identifying decay cavities in tuberculomas, X-ray CT enhanced the effectiveness of a study from 17 to 36% as compared with the results obtained at AXS and LT that revealed specific changes in the intrathoracic lymph nodes and inoculation foci in the lung. The nature and scope of elective operations were reconsidered from the results of X-ray CT. PMID- 19565812 TI - [Day tuberculous exudative pleurisy: morphological and morphometric aspects]. AB - Visceral pleural pathomorpholodical changes were studied in 50 patients. Group 1 included 25 patients with isolated tuberculous pleurisy; Group 2 consisted of 7 patients with tuberculous exudative pleurisy in the presence of a specific process in the lung; a comparison group comprised 18 who had not died from diseases of the lung and pleura. The exudative productive type of specific pleural inflammation was prevalent in 15 (60%) and 4 (57.1%) patients, respectively. In Group 1, the pleural thickness was 144.0 +/- 3.3 microm, which was 8.3 times greater (p < 0.01) than that in the comparison group (17.35 +/- 4.78 microm). In Group 2, the visceral pleural thickness was 151.1 +/- 3.4 microm, which was 8.7 times greater (p < 0.01) than that in the comparison group and 1.05 times greater than that in Group 1. The study has established that in both Group 1 and Group 2, exudative pleurisy is accompanied by early visceral pleural fibrosing with a tendency to be more pronounced in Group 2 patients, which raises the question as to whether it is necessary to use resolution therapy at the early stages of treatment, to inject isoniazid lymphotropically, and to apply physical methods to prevent pleural cavity obliteration. PMID- 19565813 TI - [Serum nitric oxide level in the assessment of systemic inflammation in patients with drug-resistant pulmonary tuberculosis]. AB - Two hundred and forty-three patients with active pulmonary tuberculosis were examined. Their sera were tested for the level of stable nitric oxide (NO) metabolites, by using the Griess reagent after previous reduction of nitrates to nitrites by a copper-impregnated cadmium reducer. The frank active pulmonary tuberculosis was ascertained to follow the lower serum levels of NO metabolites. The serum NO level did not correlate with inflammatory markers, but reduced when the process was recurrent or chronic. By taking into account the role of NO in the performance of the body's different systems, its serum reduction in patients with pulmonary tuberculosis should be probably referred to as the manifestations of metabolic decompensation as the suppressed endothelial NOS activity that determines the level of NO in circulation. PMID- 19565814 TI - [Antituberculosis humoral immunity in patients with tuberculosis, HIV infection concurrent with tuberculoses]. AB - Enzyme immunoassay (EIA) was used to study the diagnostic value of PTAT circulating in the free and attached state as part of serum circulating immune complexes (CIC) in 34 patients with tuberculosis (TB), 38 with concomitant HIV/TB infection, and 92 with HIV infection without active tuberculosis. A humoral immunological response as circulating PTAT and PTAT conjugated in specific CIC depends on both the form and degree of pulmonary tuberculosis and may be employed to diagnose a disease running as both an independent entity and in the concomitant HIV/TB infection. The use of specific CIC yields additional information on the presence and circulation of mycobacterial antigenic components, which should be borne in mind at the early stages of the disease. EIA diagnosis of pulmonary tuberculosis from the significant titer of PTAT is relatively effective at the stages with infiltration and it ascertains destructive changes before their detection on X-ray and tomographic films. PMID- 19565815 TI - [Detection of mutations in codon 306 of the embB gene for molecular genetic characterization of clinical Mycobacterium tuberculosis strains]. AB - A total of 254 Mycobacterium tuberculosis strains were used in the study. Among them, there were 183 ethambutol (EMB)-resistant strains, 13 multidrug resistant ones, but EMB-sensitive, and 39 strains sensitive to rifampicin (RIF), isoniazid (INZ), and EMB. All the strains were analyzed for genetic changes in three loci: embB306, rpoB, and katG/inhA promoter, which were associated with the formation of resistance to EMB, RIF, and INZ, respectively. The Mycobacterium tuberculosis strains were obtained from pulmonary tuberculosis patients living in the Central Region of the Russian Federation. Resistance to RIF, INZ, and EMB was revealed by the absolute concentration test. The inhibitory concentration (IC) of EMB was determined for all the strains. Genetic changes in the above loci were estimated by mini-sequencing, followed by mass-spectrometry recording MALDI-TOF products. The relative low frequency of embB306 mutations was observed among the EMB resistant strains (about 41.5%). Mutations in codon 306 were detected only in strains with EMB IC > or = 2 mg/L. A statistical significant association was found between the frequency of embB306 mutations and the multidrug resistant phenotype. A combination of these mutations with the traditional genetic markers of multidrug resistance may be used for the more effective detection of multidrug resistant strains. PMID- 19565816 TI - [Pathomorphology of specific inflammation in silicotuberculosis]. AB - Silicotuberculosis is an independent disease that arising in patients with silicosis from exacerbation of old tuberculous foci in the lung, less frequently in the lymph nodes and other organs. In silicotiberculosis, there are common secondary tuberculosis forms that are located mainly in the lung, which are rarely accompanied by a rapid progression. Characteristic morphological signs of early, nodal and nodular silicosis are observed. Silicotuberculosis is an independent disease that arises in patients from an exacerbation of old tuberculous foci in the lung, less frequently in the lymph nodes and other organs. In silicotuberculosis, there are usually secondary tuberculosis forms that are located mainly in the lung, which are rarely accompanied by a rapid progression. The characteristic morphological signs of early, nodal, and nodular silicosis are observed. Thoracic and abdominal lymph nodes, lymphatic and blood vessels, the bronchi and pulmonary surfactant system were explored. In silicosis, tuberculosis, and silicotuberculosis, silicon levels and spodograms of some visceral organs were studied and trace elements were determined in the lung. PMID- 19565817 TI - [Pathomorphism of pulmonary tuberculosis under the present conditions (according to autopsy data)]. AB - Intraoperative (n = 290) and autoptic (n = 246) materials obtained from tuberculosis hospitals in 2005-2007 were examined to study the pathomorphism of pulmonary tuberculosis. The study revealed an increase in the number of patients with acute pulmonary tuberculosis. Fibrocavernous tuberculosis halved as compared to the 1980s. Incapsulated pulmonary tuberculosis is prevalent in the phthisiosurgical material (more than 70% of cases). The leading cause of pulmonary tuberculosis was progression of a specific inflammatory process while cardiopulmonary failure and visceral amyloidosis became rare causes of death. The higher incidence of poorly progressive pulmonary tuberculosis in males (they accounted for 85% of the number of death cases) was associated with concomitant and aggravating diseases, such as alcoholism, drug addiction, and HIV infection (68.5% of cases). PMID- 19565818 TI - [Assessment of the training of phthisiatricians in the regions of Russia]. PMID- 19565819 TI - [Leningrad blockade and its medical implications. Part 1]. AB - Medical consequences of Leningrad blockade are considered from the standpoint of modern science. Pathologies acquired by blockade survivors are regarded as extreme polymorbid conditions caused by combined effect of prolonged psychemotional stress, largest-scale famine, and hypothermia responsible for undermining vital functions. Special emphasis is laid on the role of nervous system dysregulation in the development of many diseases. The influence of physiological and pathological stress on the outcome of alimentary dystrophia is discussed in the psychosomatic context. Pathogenesis of dystrophia and Leningrad hypertension is analysed. Mechanisms of survival and rehabilitation in subjects with persistent traces of blockade pathology are considered. PMID- 19565820 TI - [Gastroduodenal diseases and Helicobacter pylori infection: overview]. AB - Brief history of the discovery of Helicobacter pylori (Hp) is presented along with data on the occurrence of this organism, its genetic properties, virulence, and influence on human organs. The role of Hp in the induction and development of various diseases (chronic gastritis, ulcer disease, gastric cancer and maltoma) is discussed. Eradication therapy recommended by the Maastricht consensus 1-3a and alternative strategies are described with emphasis on their negative consequences. Problems of Hp resistance to medication, reinfection, and diagnosis of Hp-associated conditions are considered along with results and prospects of their control. PMID- 19565821 TI - [Diagnosis and differential diagnosis of primary hyperaldosteronism]. AB - This literature review concerns diagnosis and differential diagnosis of primary hyperaldosteronism, a frequent cause of arterial hypertension. It describes different phases of diagnosis of this condition including main medicamentous tests and instrumental methods as well as nosological entities from which primary hyperaldosteronism needs to be distinguished. PMID- 19565822 TI - [Lipid peroxidation and antioxidative protection in patients with coronary heart disease]. AB - Total cholesterol as well as cholesterol of low and high density lipoproteins, lipid peroxidation products (LPP), such as dienic conjugates and compounds reacting with thiobarbituric acid, were measured in a single blood sample from each of the 295 patients with CHD and 20 apparently healthy subjects. FC II and III angina was diagnosed in 142 and 151 CHD patients respectively. Certain samples were used to additionally detect homocysteine, activity of anti-oxidative enzymes (superoxide dismutase and glutathione peroxidase) in erythrocytes and ceruloplasmin/transferrin anti-oxidative system in plasma. Patients with FC III angina had significantly higher LPP levels but lower activity of tissue and plasma anti-oxidative systems than patients with FC II angina or healthy subjects. Patients with stable forms of CHD showed correlation between lipid composition, LPP contents, homocysteine level, and activity of anti-oxidative enzymes. PMID- 19565823 TI - [Heart rate variability in chronic pulmonary heart]. AB - Early signs of dysregulation of the cardiovascular system were revealed in 265 patients by studying peripheral vegetative regulation of pacemaker activity of the sinus node by rhythmocardiography with the use of a high-resolution KAP-RK-01 Mikor system (Chelyabinsk). Cardiac rhythm variability suggested impaired peripheral vegetative regulation of sinus node chronotropic function and functional insufficiency of pacemaker cells. Early symptoms of chronic pulmonary heart in patients with persistent asthma were associated with autonomous cardioneuropathy that correlated with structural changes in the heart. Peripheral vegetative dysregulation and functional insufficiency of pacemaker make up a pathogenetic component of cardiac disorder in patients with bronchial asthma. It is concluded that autonomous cardioneuropathy with marked suppression of sympathetic and parasympathetic regulation in the sinus node is an early manifestation of pulmonary heart development. PMID- 19565824 TI - [Pathogenesis of arterial hypertension in patients with primary hypothyroidism]. AB - The aim of this work was to study the role of hypovolemia on pathogenesis of arterial hypertension (AH) in patients with primary hypothyroidism (PH) and specification of indirect signs indicative of increased circulating blood volume. 217patients with PH and AH (mean age 53.3 yr) received substitution therapy with sodium levothyroxin and standard hypotensive therapy with ACE inhibitors. All the patients allocated to 3 groups were examined by 24 hour Holter monitoring and measurement of serum TSH and free T4. Group 1 (n = 74) were patients with compensated PH, group 2 (n = 57) comprised patients with subclinical hypothyroidism, group 3 (n = 86) were patients with decompensated hypothyroidism. The latter group was dominated by non-dippers, the patients showed elevated systolic and diastolic AD, its small reduction and high variability at night time. These clinical and functional signs are supposed to reflect hypovolemia in patients with PH and concomitant AH. PMID- 19565825 TI - [The role of free and esterified fatty acids in pathogenesis of metabolic syndrome]. AB - Patients with insulin resistance and those with metabolic syndrome (MS) in the absence of alteration of glucose-insulin homeostasis were found to have abnormal composition of plasma free fatty acids (FA) and lipid FA in erythrocytes. Patients with metabolic syndrome and different glucose-insulin homeostasis accumulated polyenic FA and lost saturated FA in plasma; saturated FA in their erythrocytes increased while polyunsaturated FA decreased. These findings suggest disturbed receptor-mediated active transport of FA and the predominance of passive absorption of free FA by cells as a cause of insulin resistance and hyperinsulinism. PMID- 19565827 TI - [Eradication therapy and processes of proliferation and apoptosis in the stomach of patients with duodenal ulcer]. AB - A total of 172 subjects were enrolled in the study including 100 with duodenal ulcer (DU), 30 with chronic non-atrophic gastritis (CNG), 30 with chronic atrophic gastritis (CAG), and 12 practically healthy controls. All DU, CNG, and CAG patients had Helicobacter pylori (Hp) infection confirmed by morphological study of gastric mucosa. Patients with DU were divided into 2 groups matched for age, endoscopic, morphological, and immunohistochemical characteristics. In group 1, Hp was eradicated by traditional therapy with omeprazole (20 mg), clarithromycin (500 mg), and amoxicillin (1000 mg) twice daily for 7days. This treatment in group 2 was supplemented by dibicor (500 mg twice daily). Thereafter, patients of group 1 continued to receive omeprazole for 6 weeks followed by maintenance therapy; those of group 2 received omeprazole plus dibicor for 6 weeks and maintenance therapy. CNG and CAG patients and healthy subjects served as controls. Fibrogastroduodenoscopy was performed after 2 and 4 weeks. CNG and CAG patients were examined for the presence of KI-67, BCL-2 regulators and apoptotic activity of epitheliocytes from antral mucosa before and 6 weeks after eradication therapy. It is concluded that its combination with dibicor significantly contributes to Hp elimination, shortens duodenal scarring time, improves KI-67, BCL-2 expression, and stimulates apoptotic activity of epitheliocytes compared with traditional omeprazole treatment. PMID- 19565826 TI - [Osteogenic factor in pathogenesis of vertebrogenic cardialgia]. AB - A total of 136 patients with clinical symptoms of thoracic osteochondrosis and reflex vertebrocardiac disorders were examined. 100 of them presented with vertebrogenic cardialgia and CHD, 36 had no CHD. 56 patients with clinical symptoms of thoracic osteochondrosis and CHD received traditional therapy and a course of intraosseous blockade (IOB). 56 patients with clinical symptoms of thoracic osteochondrosis and CHD received traditional therapy and a course of paravertebral blockade (PVB). 36 patients with clinical symptoms of thoracic osteochondrosis without CHD received a course of IOB. Efficiency of therapy was assessed from dynamics of pain syndrome parameters, data of the visual analog scale, McGill and body diagram questionnaires, changes of cardiac function and heart rhythm recorded by 24-hr Holter monitoring. Therapeutic efficiency of IOB in patients with vertebrogenic cardialgia with and without CHD was higher than that of PVB in patients with clinical symptoms of thoracic osteochondrosis and CHD. It is concluded that osteogenic factor plays a key role in the mechanism of development of vertebrogenic cardialgia. PMID- 19565828 TI - [24-hour monitoring of electrocardiogram, arterial pressure, and psychovegetative disorders during acute peptic ulcer hemorrhage in patients with hypertensive disease]. AB - 65 patients with hypertensive disease (HD) and without it were examined in the acute period of gastroduodenal ulcer hemorrhage (UH) by 24 hour ECG monitoring, measurement of arterial pressure (AP), and evaluation of the severity of psychovegetative syndrome. Control groups comprised 20 patients with ulcer disease (UD), 28 with HD, and 12 with UD + HD. Patients of the latter group, similar to normotensive ones, suffered suppressed function of the sinus node in addition to functional changes in myocardium including a change of ST-T segment. Circadian rhythm of AD was unrelated to the presence and absence of HD. In both cases, blood loss was associated with anxiety-depressive condition and well apparent ergotropic reactions. Patients with UD and HD had more expressed emotional and personality disorders associated with impaired vegetative response. Dippers had marked sympathetic activity at rest and in vegetative reactivity tests whereas non-dippers showed ergotropic effects in similar tests. PMID- 19565829 TI - [Endothelium condition and platelet aggregation in patients with newly diagnosed type 1 diabetes mellitus and undifferentiated connective tissue dysplasia]. AB - The study included 68 patients aged 18-32 years with newly diagnosed type 1 diabetes (DM) and/or undifferentiated connective tissue dysplasia (CTD). Ten healthy subjects without clinical symptoms of CTD served as controls. Induced platelet aggregation, Willebrand factor activity, plasma thromboxane B2, 6-keto prostaglandin F1a, and endotheline-1 were measured. The results suggest relatively normal functional reserves of endothelium and platelet activity in patients with newly diagnosed DM and compensated carbohydrate metabolism. Elevated vasopressor activity and platelet aggregation in patients with DM and CTD compared with patients without CTD may be one of the mechanisms of angiopathy. PMID- 19565830 TI - [Prospects for the use of fullerenes as antioxidants in pathogenetic therapy of bronchial asthma]. AB - Bronchial asthma (BA) is a commonest respirative pathology; severe BA not infrequently has a fatal outcome. Oxidative stress is an important pathogenetic component of BA. Hyperactivity of lipid peroxidation may be due to primary genetically determined functional insufficiency of the anti-oxidative system. Recent progress in medical nanothechnologies, e.g. the use of BA fullerenes (C60) with anti-oxidative properties, opens up the possibility to obtain hydrated C60 producing no acute or chronic toxic effects and use them as therapeutic and preventive drugs for enteral and parenteral administration to humans with BA. PMID- 19565831 TI - [Prevention of neuro- and cardiotoxic side effects of tuberculosis chemotherapy with noopept]. AB - The study evaluated clinical efficiency of noopept used to prevent adverse side effects of antituberculous agents. It included 60 patients with newly diagnosed respiratory tuberculosis. Those in group 1 (n = 30) received 10 mg of noopept twice daily during the first month. The treatment promoted functional normalization of vegetative nervous system and antioxidative systems, reduced manifestations of anxiety, decreased frequency of adverse neuro- and cardiotoxic responses to antituberculous drugs. PMID- 19565832 TI - [Spontaneous pneumomediastinum as complication of alimentary toxicoinfection]. AB - The author reports a case of spontaneous pneumomediastinum in a previously healthy young boy resulting from alimentary toxicoinfection with frequent nausea. Adequate treatment completely eliminated symptoms of pneumomediastinum and prevented development of its complications. Published data suggesting other possible causes of spontaneous pneumomediastinum are discussed. PMID- 19565834 TI - [1st Russian congress of chronobiology and chronomedicine]. PMID- 19565833 TI - [Familial leptospirosis caused by Leptospira grippotyphosa]. AB - Leptospirosis is a zoonotic anthropurgic infection that occurs in the human population as sporadic cases or affects groups of individuals. Outbreaks are possible at any territory with a landscape harbouring natural reservoirs of Leptospyras and inhabited by small moitsaure-loving rodents. Leptospirosis peak season is summer. Specific diagnostic features and laboratory characteristics of 3 cases of leptospyrosis in a single household area are described. PMID- 19565835 TI - [The influence of physician's personality on professional activity. Part 2]. PMID- 19565836 TI - Plan demographics, participants' saving behavior, and target-date fund investments. AB - This analysis explores (1) whether plan demographic characteristics would affect individual participant contribution rates and target-date fund investments and (2) equity glide paths for participants in relation to plan demographics by considering target replacement income and its success rate. PLAN DEMOGRAPHIC CHARACTERISTICS IN PARTICIPANT CONTRIBUTION RATES: This study finds empirical evidence that 401(k) plan participants' contribution rates differ by plan demographics based on participants' income and/or tenure. In particular, participants in 401(k) plans dominated by those with low income and short tenure tend to contribute less than those in plans dominated by participants with high income and long tenure. Future research will explore how participant contribution behavior may also be influenced by incentives provided by employers through matching formulae. PLAN DEMOGRAPHIC CHARACTERISTICS IN TARGET-DATE FUND INVESTMENTS: The study also finds empirical evidence that participants' investments in target-date funds with different equity allocations differ by plan demographics based on participants' income and/or tenure. In particular, target date fund users with 90 percent or more of their account balances in target-date funds who are in 401(k) plans dominated by low-income and short-tenure participants tend to hold target-date funds with lower equity allocations, compared with their counterparts in plans dominated by high-income and long tenure participants. Future research will focus on the extent to which these characteristics might influence the selection of target-date funds by plan sponsors. EQUITY GLIDE PATHS: Several stylized equity glide paths as well as alternative asset allocations are compared for participants at various starting ages to demonstrate the interaction between plan demographics and equity glide paths/asset allocations in terms of success rates in meeting various replacement income targets. The equity glide path/asset allocation providing the highest success rate at a particular replacement rate target will vary with the assumed starting date of the participant (see Figure 17). Given the highly stylized nature of the simulations in this Issue Brief it is important to note that the results are not intended to provide a single equity glide path solution in relation to plan demographics. Instead, they serve as a framework to be considered when plan sponsors make a selection concerning which target-date funds to include in their plan. IMPORTANCE OF PARTICIPANT CONTRIBUTION RATES: This analysis finds that although target-date funds with different equity glide paths affect the retirement income replacement success rate, participant contribution rates corresponding to different plan demographic characteristics have a stronger impact. AUTO FEATURES OF THE PPA: This Issue Brief provides a stylized study using observed contribution rates as of the 2007 plan year. However, with the passage of the Pension Protection Act of 2006 and its likely impact on plan design in the future (increased utilization of automatic enrollment and automatic contribution escalations), it is likely that contribution rates among the participants may become more homogenous. In such a scenario, it may be more likely that a single equity glide path would meet a wide range of demographic profiles. PMID- 19565837 TI - [Studies on heredity rule of the first genealogy regarding fatal familial insomnia in Henan province]. AB - OBJECTIVE: To investigate the epidemiological, genealogic characteristic, familial history of the families with fatal familial insomnia, its clinical and pathological features as well as the heredity rule of related genes. METHODS: 135 familial members of 7 eras were studied. Vein blood samples from patients as well as from some familial members were collected. PRNP gene was studied with PCR, its serial was determined and then authenticated with Nsp I . Brain tissue was obtained for neuropathological test and PrP(Sc) test with Western blot method. RESULTS: Clinical symptoms of the 2 diagnosed cases were typical. 11 familial members died of similar neural disease. 32 samples of their familial members, codon at D178N of PRNP of 11 members was mutated, with mutation rate as 34.38% while D129N showed as methionine. Brain tissue of both probands denaturalized into spongiform and the nerve fiber was absent but PrP(Sc) protein was identified. CONCLUSION: Genealogy was described in the family with fatal familial insomnia since the patients had typical clinical symptoms and pathological characteristics. It seemed necessary to confirm cases of fatal familial insomnia and their genealogy with epidemiological data and to investigate its gene characteristics as well as with neuropathological and Western blot tests. PMID- 19565838 TI - [Study on time-space clustering regarding the distribution of visceral leishmaniasis in Kashgar Region, Xinjiang]. AB - OBJECTIVE: To probe time-space clustering on the distribution of visceral leishmaniasis (VL) in Kashgar Region. METHODS: Based on the geographic information system, a Poisson model of time-space statistical software was applied to analyze data over the past 11 years in the Kashgar Region. Zones with clustering phenomenon were conformed by geographic location and remote sensing images. RESULTS: There existed three high risk clustering zones and corresponding time frames of VL in Kashgar Region. The center location of zone A was located in E 76.08 degrees, N 39.52 degrees, with radius as 6.58 km. The high risk time frame was from January 1st of 1999 to December 31st of 2003. Within the zone and time frame, the relative risk (RR) of VL incidence was 45.98 times higher than those outside the scope (P < 0.0001). Zone B' s center location was at E 79.20 degrees, N 39.91 degrees, with the radius as 4.93 km. Its high risk time frame was from January 1st of 2002 to December 31st of 2006. Within the zone and time frame, the RR of VL incidence was 9.58 times higher than those outside of the scope (P < 0.0001). Zone C' s center location was in E 76.23 degrees, N 39.40 degrees, and the radius was 7.63 km, with the high risk time frame from January 1st of 2000 to December 31st of 2004. Within the zone and time frame, the RR of VL incidence was 5.18 times higher than the one from outside of the scope (P < 0.0001). CONCLUSION: The incidence of VL in Kashgar Region was non-randomly distributed while there existed obvious time-space clustering, with all of three high risk clustering zones located in oasis area where appeared the focus area for control and surveillance of VL. PMID- 19565839 TI - [Study on HIV/AIDS related high risk behaviors among unmarried men who have sex with women]. AB - OBJECTIVE: To explore the risk of sexual behaviors related to AIDS among unmarried men who often have sex with women. METHODS: Study objectives were recruited through target sampling and anonymous questionnaires were adopted. Men' s sexual behaviors were compared with each other between the group of men who always had sex with women (group A) and who never had sex with women (group B) in the past 6 months. RESULTS: The average numbers of same-gender partners among group A who acted as the male role in the past 6 months and who had anal intercourse in the last month were 5.64 and 3.09 respectively, which were both higher than those of group B's (P < 0.05). The rate of condom use among men in group A when having sex with men was 81.94%. The percentages of condom use among men in group A who having had oral or anal sex with male sex buyers last time were 15.38% and 70.27% respectively. They were all higher than those of group B' s (P < 0.05 or 0.01). The rates of men in group A who were male sex workers and who having had sex with unknown men in gay places during the past 6 months were 27.59% and 61.70% respectively. Among group A, the percentages of men who had had sex with unknown men during the last time and who had had sex with nonlocal male partners during the past year were 43.06% and 40.28% respectively. They were all higher than those of group B's (P < 0.01). The percentage of condom use among men in group A when having had sex with women during the past 6 months was 68.94%, but only 25.96% of the men used condom regularly. The rate of condom use in men from group A when having sex with women during the last sexual intercourse was 65.45%. CONCLUSION: HIV high-risk sex behaviors were more ubiquitous among unmarried MSM who always had sex with women, suggesting that different interventions be developed to prevent them from transmitting HIV from MSM to women or other men. PMID- 19565840 TI - [The impact of childhood sexual abuse on the development of AIDS related high risk behaviors and psychological appearances among men who have sex with men]. AB - OBJECTIVE: To study the AIDS related high risk behaviors and psychological appearances among men who have sex with men (MSM) who ever experienced childhood sexual abuse (CSA). METHODS: Target sampling for a cross-sectional study was developed and valid anonymous questionnaires were adopted to compare the differences of high risk behaviors related to AIDS and psychological appearances between those with or without CSA experiences among 2147 MSM from nine cities. RESULTS: Compared to corresponding ones without CSA experience, CSA group had a significant larger numbers in the following events: total sexual partners, anal sex episodes with same sex, female sexual partners and anal sex in the previous six months, with the figures of median as 20.0, 10.0, 3.0, 3.0 respectively. In the previous year, 30.8% of them had ever participated in 'group sex', 19.2% ever exchanged money for sex, 36.7% bled while having sexual intercourse, 37.3% had sex with male partners away from his own region. All the above said figures were higher than non-CSA group, with significant differences. It also appeared that CSA experience had an impact on significant lower rate of condom use (67.3%) in the last anal sex. Those with CSA experience had more psychological problems which appeared as: 75.6% considered they would suffer from serious discrimination if their sexual orientation ever disclosed, 34.7% had a strong intention of suicide and 24.3% ever having had suicidal attempts. The differences of the two groups showed statistical significance. CONCLUSION: CSA experience not only increased the number of AIDS related high risk behaviors in adulthood, but also had negative impact on their psychological appearances. It is of urgent need to carry out psychological intervention approaches to target on MSM with CSA experiences while childhood sexual education and rights assurance towards juvenile population should also not be neglected. PMID- 19565841 TI - [Study on the effect of intervention about acquired immunodeficiency syndrome among men who have sex with men]. AB - OBJECTIVE: To study the outcomes of AIDS intervention programs and to provide scientific evidence for developing pertinent strategy on intervention among men who have sex with men (MSM). METHODS: MSM were recruited through snowballing and investigated by face to face interview in 2006 and 2007 respectively. SPSS 12.0 was used to compare the change of cognition about AIDS, sexual behavior, prevalence rates of HIV and Syphilis before and after the intervention program among the population under study. RESULTS: The cognition about AIDS among MSM was obviously improved after the intervention with the rate increased from 74.3% to 82.4% (P = 0.01). The rate of last time condom use among MSM increased from 56.4% to 65.5% (P = 0.00). The rate of consistent condom use during six months among MSM increased from 31.8% to 41.9% (P = 0.00). The rates of both condom use during commercial sex with men and with women did not change much among MSM. The prevalence rates of HIV in 2006 and 2007 were 10.4% and 10.8% and of syphilis in 2006 and 2007 were 9.3% and 7.3% respectively which were not significantly different between before and after the intervention. CONCLUSION: The cognition about AIDS among MSM was improved obviously. However, the rate of consistent condom use was still low, reflecting the segregation phenomenon between their behavior and cognition. All our findings implied that it was crucial to carry out AIDS prevention and control programs. PMID- 19565842 TI - [Study on the school-related-factors of attempted suicide among rural middle school students]. AB - OBJECTIVE: This study was undertaken to examine possible relationship between attempted suicide and underachievement, bullying, low life satisfaction and low self-concept at school. METHODS: An anonymous self-report survey assessing demographic characteristics and the major risk factors of teenage attempted suicide was completed by students from 16 middle schools in grades seven to twelve in 4 counties of Anhui province (age 10 to 21 years). An anonymous questionnaire was used to rate attempted suicide, bullying involvement and learning performance. Attempted suicide was defined as: experiencing specific suicide actions at least one time during the 12 months preceding the survey. Multidimensional Students' Life Satisfaction Scale and Children' s Self-concept Scale were used to evaluate satisfaction and self-conscience on and at respectively. In total, 10 894 respondents substantially completed the survey. Multiple logistic-regression analyses, controlling for socio-demographic variables, was used to analyze if underachievement, bullying, low school life satisfaction and low children' s self-conscience at school had been risk factors. RESULTS: 629 participants (5.8%) reported having made at least one attempted suicide within the last 12 months. Students being underachieved were significantly having more attempted suicide events than those excellent students (chi2 = 11.39, P = 0.023). Students being both bully-victims and practiced bully were significantly more than those being only practiced bully (28.7% vs. 15.8% , P < 0.001), being victims (28.7% vs. 10.6% , P < 0.001) or having neither of them (28.7% vs. 4.8%, P < 0.001). Results from multiple logistic regression analyses showed that underachievement, bullying, lower school life satisfaction and low self conscience were risk factors for attempted suicide. CONCLUSION: Data from this study confirmed that school bullying and children' s self-conscience at school were significantly associated with attempted suicide among rural middle school students in Anhui province. It is of importance to improve the school' s environments to reduce the risk of attempted suicide among this group. PMID- 19565843 TI - [Distribution regarding tendency on personality disorder among college students in Shijiazhuang city]. AB - OBJECTIVE: To survey the prevalence of tendency on tendency of personality disorder among college students. METHODS: By means of stratified cluster sampling, 498 students from 6 colleges in Shijiazhuang city and 204 students from 3 colleges in Beijing were studied through 'personality diagnostic questionnaire revised UPDI'. RESULTS: The incidence rates on dependent personality (2.81%), histrionic personality (2.41%) and borderline personality (2.21%) were higher than obsessive-compulsive personality (0.40%) and schizoid personality (0.60%). The prevalence of personality disorder tendency was related to sex, major and years in college, blood type as well as their origins (from urban or rural). The overall incidence of personality disorder was 28.31% while the incidence rates of personality deviation and serious personality disorder tendency were 17.07% and 11.24% respectively. The incidence in males was higher than that in females. There appeared differences in dissociative personality, avoidant personality, paranoid personality, obsessive-compulsive personality, histrionic personality and narcissistic personality on people with different blood types. The scores of the city students were higher than that of the students from the rural areas regarding paranoid personality, dependent personality and narcissistic personality. Differences were also noticed between freshmen and students from other levels in the incidence rates on the tendency of avoidant personality disorder. CONCLUSION: There were different incidence rates on the tendency of personality disorder among college students that related to sex, level in college and the origins where they were from (urban or rural). PMID- 19565844 TI - [What is the prevalence of smoking in China]. AB - OBJECTIVE: To determine the accuracy of prevalence data sets on tobacco use so as to measure the risk of tobacco use and the impact of tobacco control in China. METHODS: Three published data sets on nation-wide survey were reviewed and compared. Two principles were applied to determine the accuracy of the data on prevalence: i ) The estimated consumption of cigarettes based on the current prevalence rate on smokers should have been close to the actual cigarette consumption level; ii) change on the annual prevalence of male current smokers should be around 1% in China, since the international experience on the prevalence of current smokers tended to decrease at a rate of around 1% per year in the presence of comprehensive tobacco control strategies. RESULTS: The differences between the estimated cigarette consumption and the actual cigarette consumption for the three surveys were 51.07 billion through Behavior Risk Factor Surveillance System (BRFSS 2002), 62.94 billion through the Chinese Citizen Nutrition & Health Survey (CCNHS 2002), and 217.11 billion through the China Health Service Survey (CHSS 2003). In comparison with the national tobacco use survey in 1996, the prevalence of male current smokers apparently dropped by 0.9% in BRFSS 2002, 2.2% in CCNHS 2002 and 2.0% per year in CHSS 2003. Thus, the prevalence of current smokers in BRFSS (2002) was more reliable, comparing to the results from the other two surveys. CONCLUSION: The prevalence of current smokers as determined by the BRFSS should be used to reflect the epidemic of tobacco use when implementing the FCTC in China. However, the reporting prevalence rates of tobacco use were different in the different surveys regarding tobacco use, suggesting that the capacity of surveillance on tobacco control should be strengthened, including the standardization of definitions on 'ever-smoker' and 'current smoker' , as well as on standardized questionnaire, sampling strategy and the process of data analysis, quality of field work etc. Precise estimation of prevalence appears to be the key point for understanding how many current smokers so as to develop control policy, including setting up 'quit' clinics and evaluating the impact of tobacco control programs. There is an urgent need to establish a national standardized surveillance system to monitor the tobacco epidemics. PMID- 19565845 TI - [The impact of air temperature variation on the visits to emergency room in Shanghai]. AB - OBJECTIVE: To assess the association between air temperature and emergency room visits among patients covered by medical care program from 'third-grade' hospitals in Shanghai. METHODS: Generalized additive model (GAM) was used to analyze time series, and AR(P) was used to deal with auto correlation of time series. After controlling factors as both medium-term and long-term trends, day of the week, vocation, typical pneumonia and pollutants, the association between air temperature and emergency room visits in virtue of quadratic curve and differential coefficient principle were estimated. RESULTS: When air temperature was below 14.71 degrees C, the increase of 95% confidence interval to relative risk in corresponding emergency room visits along with 1 degree C increase of air temperature, was less than 1. However, when air temperature was above 19.59 degrees C, the relative risk's 95% confidence interval was greater than 1. When air temperature varied at the range of 14.71 degrees C-19.59 degrees C, the 95% confidence interval of the relative risk would include 1. Hence, air temperature range between 14.71 degrees C-19.59 degrees C, was called the optimum temperature range. CONCLUSION: Our findings indicated that the current air temperature had an acute impact on the number of emergency room visits among patients covered by medical care program visiting those third grade hospitals in Shanghai. PMID- 19565846 TI - [Study on the determinants regarding malaria epidemics in Anhui province during 2004-26]. AB - OBJECTIVE: To analyze the determinants of malaria in Anhui province during the year of 2004-2006. METHODS: A GIS-based database of malaria at the township scale of Anhui province was established, using remote sensing and spatial analysis technologies. Methods on statistical analysis, principal component analysis combined with logistic regression analysis were synthetically used to analyze the association between malaria and environmental factors. RESULTS: Malaria epidemics in Anhui province during 2004-2006 mainly occurred in the northern districts of Huai River, and the epidemics had become more serious yearly. The determinants of malaria at the township scale mainly included factors as temperature, rainfall, normalized difference vegetable index and elevation. If the lowest temperature in a year, which appeared the fourth principal component of the temperature index series, increased by one unit, the probability of incidence of malaria would decrease by 33%. If the total annual rainfall, which was ihe first principal component of the rainfall index series, increased by one unit, the probability would decrease by 27%. If the elevation increased by 10 meters, the probability would decrease by 2%. However, he relationship between Normalized Difference Vegetable Index (NDVI) and the probability of incidence of malaria was different. If the NDVI had a one unit increase, the probability would increase 3.28 times. CONCLUSION: The northern districts of Huai River during 2004-2006 appeared to be a new spatio-temporal cluster when reemergence of malaria epidemics had occurred in Auhui province since 2000. Terrain and physiognomy, nature and circumstances factors, such as temperature and rainfall had affected the incidence rates of malaria. Our research data from Anhui province would provide some important references to the discovery of main reasons on the reemergence of malaria epidemics since 2000, especially in the central geographic areas of China. PMID- 19565847 TI - [Epidemiological analysis of imported cases of dengue fever in Guangdong province and Hong Kong during 2004-2006 in China]. AB - OBJECTIVE: To analyze the epidemiological characteristics of imported cases of dengue fever in Guangdong province (GD) and in Hong Kong (HK) during 2004-2006 to provide evidence for further cooperation in the prevention and control programs on dengue fever in the two places. METHODS: Descriptive statistical analysis was performed on data obtained from dengue fever surveillance and reporting network in GD and from Centre for Health Protection, Department of Health, HK. RESULTS: Both from GD and HK 44 and 93 imported cases of Dengue fever were reported during 2004-2006. Most patients from GD acquired their infection from Singapore (13 cases), Indonesia (9 cases) or Cambodia (6 cases) while patients in HK mainly were imported from Indonesia (31 cases), the Philippines (16 cases) and Thailand (15 cases). The peak seasons of the two places were both from July to September. During the non-peak season period, the number of cases in Hong Kong was higher than that in Guangdong. Male/ female ratio was 1.2:1 in GD and 1.1:1 in HK. Age of patients in GD appeared to range from 6-80 years, with 63.6% (28/44) of them aged 20-39 years. 40.9% (18/44) of the cases were engaged in business, services, housework or unemployed. Those cases in HK were between 10-72 years of age, with 63.6% (28/ 44) of them aged 20-39 years while 47.3% (44/93) of the patients were with the occupation of business, services and industry. More cases in GD had a onset of disease before entering the border (27:17) than the cases in HK (35:57). The average time interval between onset and diagnosis were 7 and 9 days for GD and HK respectively. CONCLUSION: Frequent travel between Southeast Asia in summer among the working class appeared to be the main factor, causing imported cases of dengue fever in GD and HK. It is crucial to provide health education targeted at these high risk groups in order to prevent importation of dengue fever in the two areas. PMID- 19565848 TI - [Genetic characteristics of enterovirus 71 isolated in Beijing, 2006-2008]. AB - OBJECTIVE: To sequence and analyze the VP1 region of isolated enterovirus from different sources in Beijing, 2006-2008. METHODS: 9 EV71 were selected from the isolates identified through the specimen of human hand foot mouth disease (HFMD), acute flaccid paralysis (AFP) and healthy children in Beijing, 2006-2008. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to amplify and sequence the whole VP1 gene of enterovirus. Phylogenetic tree was constructed, with the means of nucleotide homology and distance between/within groups analyzed. RESULTS: The 9 selected strains were clustered with C4 subgenotype reference strains in Phylogenetic tree and showed high nucleotide acid identity (92.1%-93.9% ) in nucleotide homology analysis, and had higher homology than C1, C2, C3 subgenotype reference strains (88.8%-89.5%, 89.4%-90.0% and 88.4%-89.3%, respectively). High homologous (95.9%-100.0%) was noticed between the isolated stains from three different sources, but low homologous (93.3% -93.9%, 92.1% 92.9%, respectively) showed between the isolated stains and C4 reference strains isolated in 1998. There appeared larger variations between groups in C4 subgenotype when analyzing the distance between groups means, especially between the reference strains and isolated strains (D = 0.052-0.071). CONCLUSION: The EV71 isolated in Beijing, from 2006 to 2008 also appeared to be C4 subgenotype and there was no significant difference found in the whole sequence of VP1 gene of the strains isolated from different regions, sources, or under different diseases occurred in the same period. There were more nucleotide variations and more chances for the presence of new subgenotype, suggesting that it is necessary to strengthen the surveillance program on EV71 isolates. PMID- 19565849 TI - [Isolation and identification of arboviruses from mosquito pools in some regions of Liaoning province, China]. AB - OBJECTIVE: To isolate and identify arboviruses from mosquito pools in some regions of Liaoning province. METHODS: Mosquitoes were collected from Shenyang, Yingkou, Panjin, Jinzhou and Dandong cities of Liaoning province in 2006. Viruses were isolated by inoculating the specimens onto C6/ 36 and BHK-21cells. The new isolates were identified using serological and molecular biological methods. RESULTS: 5410 mosquitoes were collected from the five cities in total. Three isolates produced CPE in C6/ 36 cell and five isolates produced CPE in both C6/36 and BHK-21 cell. Three isolates (LN0684, LN0688 and LN0689) were identified as Banna virus and one isolate (LN0636) was identified as Getah virus. Phylogenetic analysis showed that the three Banna virus strains were clustered into the same evolution branch as the other Chinese isolates. The identity of nucleotide sequence was between 91.2% and 94.7%, compared with other Banna virus strains. The new isolated Getah virus was clustered into the same branch with the strain of South Korea (swine). The identity of nucleotide sequence was 99.2%, when comparing with the strain of South Korea and was 95% to 99% with the strains from Russia, mainland of China and Taiwan region. Conclusion Eight virus isolates, including three Banna virus, one Getah virus and four unknown virus strains were isolated from mosquitoes in Liaoning province. Banna virus and Getah virus were reported for the first time in Liaoning province, while Getah virus showed the highest nucleotide homology with the South Korea strains. PMID- 19565850 TI - [Study on the pathogen of plague in Sanjiangyuan area in Qinghai province]. AB - OBJECTIVE: To study the biological characteristics of Yersinia pestis and to develop prevention and control program on plague in Sanjiangyuan areas, Qinghai province. METHODS: To identify the biologic types and molecular biological features of Y.pestis isolated in Sanjiangyuan area from 1954-2007. RESULTS: Among the 411 strains of Y. pestis, 12 strains belonged to the microtus type Y. pestis with denitrification (-) and donkey-hide gelatin carbohydrate (-) and glycerine (+). 399 strains belonged to classic type Y. pestis with denitrification (+) and donkey-hide gelatin carbohydrate (+) and glycerine (+). 411 Y. pestis strains had factor F I and Pst I. Among them, VW+ strains of Y. pestis accounted for 95.13% (391/411), VW-accounted for 4.87% (20/411), Pgm(+) accounted for 80.78% (332/ 411), Pgm(+/-) accounted for 9% (37/411) and Pgm(-) accounted for 10.22% (42/411) respectively. 96.82% (213/220) of the Y. pestis strains showed strong virulence to laboratory mice while 3.18% (7/220) of the strains carried medium virulence. 90.02% of the tested Y. pestis (370/411) strains had 6 x10(6), 45 x 10(6), 65 x 10(6) plasmids. 8 types of genome were found among 80 strains of Y. pestis, with 6 of them resembling ZHOU Dongsheng' s classification. Two new genome types were found. CONCLUSION: The Y. pestis in the Sanjiangyuan area had the characteristics of plague pathogen, identified in Qinghai-Tibet plateau. It is estimated that human beings are highly susceptible to the disease which spread fast, causing serious signs and symptoms with high death rate. PMID- 19565851 TI - [Discrimination power evaluation for 45 loci of variable number tandem repeats in Mycobacterium tuberculosis strains isolated from China]. AB - OBJECTIVE: To evaluate the discriminatory efficiency of multiple loci of variable numbers of tandem repeats (vNTR) in Mycobacterium tuberculosis genome. Genotyping and identification on Chinese M. tuberculosis clinical strains were used to locate a series of high discriminated loci, so as to provide the basis for creating a standardized multiple loci VNTR analysis (MLVA) to distribute fast typing and identification on Chinese M. tuberculosis. METHODS: VNTR loci which were chosen from the website http//minisatellites.u-psud.fr/ and referenced to the genome sequence of M. tuberculosis standard strain H37Rv were tested in Chinese M. tuberculosis clinical strains and H37Rv by means of PCR. The primers were designed by DNAStar software. The repeat member of VNTR unit was estimated by the result of PCR. The discrimination power of single locus or multiple loci was confirmed by the Hunter-Gaston index. RESULTS: 45 VNTR loci were tested in 135 Chinese M. tuberculosis clinical strains and H37Rv. The discrimination power of these loci appeared different from each other, with the biggest Hunter-Gaston index as 0.814 (0.797-0.830), the smallest one as 0.015 (0.001-0.028), and there were 13 loci with which the Hunter-Gaston index was bigger than 0.5. Results showed that the discrimination power was increasing by different loci that associated with each other. The more loci that were combined, the bigger the Hunter-Gaston index was. For example, the Hunter-Gaston index of Qub11-b associated with Qub18 was 0.936, by which 136 strain could be divided into 44 groups. With the combination of 9 loci including Qub11-b, Qub18, Mtub21, Rv2372, MIRU26, Qub26, Qub4156c, Qub11-a and Qub15, the Hunter-Gaston index could have reached 1 and by which the 136 swains could be divided into 136 groups, also showing that the biggest discrimination power to strain identification, viz, strain level genotype were reached. CONCLUSION: The discrimination power of different locus was different. The discrimination power of multiple loci was much more satisfied than that of single locus. It was satisfied the combine discrimination power of 9 loci including Qub11-b, Qub18, Mtub21, Rv2372, MIRU26, Qub26, Qub4156c, Qub11-a and Qub15, by which the qualified typing method could gain to facilitate research on molecular epidemiology with the Hunter-Gaston index analysis. PMID- 19565852 TI - [Establishment and preliminary application of polymerase chain reaction with confronting two-pair primers for the single nucleotide polymorphisms of metabolic enzymes]. AB - OBJECTIVE: To develop a simple, accurate, rapid, economic, large-scale detection method for the detection of single nucleotide polymorphisms (SNPs) metabolic enzymes, using polymerase chain reaction with confronting two-pair primers (PCR CTPP). METHODS: The primers of CYP1A1 (A4889G), EPHX1 (A416G) and NQO1 (C609T) were designed for PCR-CTPP, and the PCR conditions were optimized. The results of genotyping were verified by DNA sequencing. The above SNPs were detected by the PCR-CTPP detection method in a randomly selected 183 healthy individuals of Han ethnicity. The genotype frequencies were analyzed and compared with people from other ethnicities. RESULTS: The allele-specific bands of CYP1A1 (A4889G), EPHX1 (A416G) and NQO1 (C609T) were successfully amplified by PCR-CTPP under the optimal conditions and the results of genotyping were consistent with DNA sequencing. Among 183 healthy Han individuals, the genotypic distributions of CYP1A1 (A4889G) , EPHX1 (A416G) and NQO1 (C609T) showed that the wild-type, homozygous variants, and heterozygotes were 103 (56.3%), 8 (4.4%), 72 (39.3%) and 142 (77.6%), 4 (2.2%), 37(20.2%), 60(32.8%), 32 (17.5%), 91 (49.7%) respectively. The distributions of genotypes were all in accordance with the Hardy-Weinberg equilibrium (P > 0.05), with statistical differences and with other ethnic populations (P < 0.05). CONCLUSION: The SNPs of metabolic enzymes can be detected by PCR-CTPP method which is simple, accurate, rapid, economic and with large scale. PCR-CTPP can be used for large scale clinical and epidemiological screening. PMID- 19565853 TI - [A meta-analysis on the association between maternal passive smoking during pregnancy and small-for-gestational-age infants]. AB - OBJECTIVE: To study the association between maternal passive smoking during pregnancy and the small-for-gestational-age (SGA) infants and to analyze independent factors on duration, in different areas. METHODS: Totally, 8 retrospective studies published in PubMed (no year limited), OVID-MEDLINE (no year limited), CNKI (from 1911 to 2008), VIP (from 1989 to 2008), and CBM (no year limited), on maternal passive smoking during pregnancy and SGA were analyzed synthetically by Meta-analysis. RESULTS: The unadjusted pooled OR value on the association between maternal passive smoking during pregnancy and SGA was 1.45 (95% CI: 1.05-2.01). Data showed that the adjusted pooled OR value was 1.76 (95% CI: 1.15-2.69). Maternal passive smoking in early pregnancy was a risk factor for SGA (OR = 1.85, 95% CI: 1.25-2.72), and so was the exposure in mid or late pregnancy(OR = 2.12, 95% CI: 1.43-3.13). No statistically significant relationship between SGA and passive smoking at home or at workplace was found. CONCLUSION: Avoiding passive smoking in pregnancy for mothers could reduce the possibility of delivering SGA infants. Other than at home or at workplace, problem of passive smoking in other places should also be brought up. PMID- 19565854 TI - [Meta-analysis on the relationship between colorectal cancer and Helicobacter pylori infection]. AB - OBJECTIVE: Analyzing all the related studies that have been published in the world to evaluate the relationship between colorectal cancer (including colorectal adenomas) and Helicobacter pylori infection by means of Meta-analysis. METHODS: To analyze the 14 references consistent with inclusion criteria by reviewing manager 4.2, calculating the odds ratio and 95% confidence interval, then drawing funnel plot. Results The total sample size is 3165, cases and controls are 1330 and 1835, respectively. The pooled OR is 1.36, and 95% confidence interval 1.01, 1.82. The graphical funnel plot appearing asymmetrical, but analyzing by sensitivity analysis and fail-safe number calculation. The results are affected by publication bias. CONCLUSION: The results of our Meta analysis show that Helicobacter pylori infection is a possible risk of colorectal cancer. PMID- 19565855 TI - [The application of support vector machine for prediction of posttraumatic stress disorder on adults in flood district]. AB - INTRODUCTION: To predict the occurrence of posttraumatic stress disorder (PTSD), using a Support Vector Machine (SVM) on adults in flood district. Diagnostic and Statistical Manuals on Mental Disorders (IV Edition) were used to examine and diagnose the victims in flood districts. Based on the forecasting model of SVM with PTSD as dependent variables and 23 influence factors of PTSD as independent variables, prediction of PTSD was conducted among the victims. After considering 23 influence factors into the prediction model, the agreement rate of prediction of the model was 88.05 percent, with sensitivity as 75.0 percent, and specificity as 89.4 percent. CONCLUSION: The prediction model based on SVM with 23 influence factors had good effect on predicting the occurrence of PTSD. PMID- 19565856 TI - [Autoregressive integrated moving average model and circle distribution analysis of stroke mortality in Tianjin]. AB - INTRODUCTION: To develop a model for forecasting the mortality of stroke in Tianjin, China. The time series of stroke mortality from 1999 Jan. to 2006 Dec. in Tianjin city were subjected. Circle distribution analysis was used to verify the trend of time concentration. Multiple seasonal autoregressive integrated moving average model [ARIMA (p, d, q) (P, D, Q) s], based on model identification, estimation and verification of parameter, and analysis of the fitting of model, was established. Most of the deaths from stroke occurred in January and had a cycle of 12 months. An ARIMA model (0, 1, 0) x (0, 1, 1)12 was established (1--B) (1-- B12) lnx(i) = 0.001 + (1--0.537 B12)epsilon(t). CONCLUSION: ARIMA & Circle Distribution analysis is an important tool for stroke mortality analysis. Potentially it has a high practical value on the surveillance, forecasting and prevention of stroke mortality. PMID- 19565857 TI - [A review of health economic evaluation on cervical cancer screening by visual inspection with acetic acid]. PMID- 19565858 TI - [Progress on hepatitis E vaccine]. PMID- 19565860 TI - [Epidemiological analysis of congenital syphilis from year 1998 to 2007 in Zhejiang province]. PMID- 19565859 TI - [Epidemiological survey on epilepsy in Kawa ethnic minority group in Yunnan province]. PMID- 19565861 TI - [Demographic profile of men who have sex with men who seek for sex mostly on internet]. PMID- 19565862 TI - [Epidemiological investigation of a gastroenteritis outbreak caused by waterborne norovirus GG I]. PMID- 19565863 TI - [A case-control study on the risk factors of hand, foot and mouth disease in children in Tianjin]. PMID- 19565864 TI - [Single-nucleotide polymorphisms and genotype combinations in calpain-10 gene of gestational diabetes mellitus]. PMID- 19565865 TI - [Effects and trends of stroke and life expectancy among older adults--from 1990s to 2000s]. AB - OBJECTIVE: To explore the experience of stroke influencing the life expectancy (LE), active life expectancy (ALE), inactive life expectancy (IALE), and the trend of life expectancy among older adults, from 1990s to 2000s in Beijing, China. METHODS: A representative sample of 3257 elderly people living in urban or rural communities in Beijing were followed up from 1990 until 2004. Their health and survival status had been surveyed every 3-5 years. Activity Daily Living (ADL) scale, recommended by WHO was used to evaluate the physical function capability of the elderly. SAS was used to estimate LE, ALE and IALE for both periods of 1992-1997 and 2000-2004 by age and by areas of residency (rural or urban). RESULTS: LE and ALE were shorter, and IALE was longer, among the elderly with stroke than those without stroke at all age groups. Functional status at baseline was also a very important factor in determining ALE and IALE. For those active at baseline, ALE in the elderly with stroke was shorter than those without. There were no differences found in IALE between those with or without stroke, but ALE was longer than IALE. For the elderly with stroke and inactive at baseline, their IALE were longer than ALE and their ALE were at low levels in all age groups. Among those with stroke and living in urban, their LE and ALE were longer than those living in the rural area. When comparing with the period of 1992-1997, both LE and ALE increased during the period of 2000-2004 in all the elderly groups, both in urban and rural areas. The largest increment occurred among those with stroke who originated in an inactive state. CONCLUSION: Stroke reduced both quality and quantity of life of the elderly. The reductions of LE and ALE were greater among the elderly with stroke in rural than in urban areas. Both LE and ALE increased from 1992-1997 to 2000-2004 among the elderly with stroke in both urban and rural areas. PMID- 19565866 TI - [A cross-sectional study on impaired fasting glycaemia and diabetes mellitus in residents from Nangang district, Harbin city]. AB - OBJECTIVE: To investigate the detection rate of impaired fasting glycaemia (IFG), the prevalence of diabetes mellitus (DM) and related risk factors, the current situation on awareness, treatment and the rate of control on diabetes mellitus in residents living in Nangang district of Harbin city, Heilongjiang province. METHODS: A cross-sectional cluster sampling was carried out in residents aged over 35 years, living in Fendou community of Harbin city. RESULTS: Data from 3017 out of 3183 residents were analyzed. In men, women and overall residents, the detection rates of IFG were 5.38%, 2.44% and 3.75%, respectively. After standardization, the detection rates became 5.41%, 2.18% and 3.59%, respectively. The prevalence rates of DM were 12.40%, 8.46%, 10.21% and 11.80%, 8.20% and 9.77%, before and after standardization. Results showed statistical difference between age, sex and levels of fasting glycaemia. Data from single factor analysis revealed that smoking, BMI, hypertension and high triglyceride were risk factors on the levels of high fasting glycaemia. Results from multivariable stepwise analysis showed that sex, age, BMI, hypertension and high triglyceride were significant factors influencing the levels of high fasting glycaemia with the OR (95% CI) is 1.546 (1.250-1.912), 1.308 (1.171-1.461), 1.038 (1.010-1.066), 1.388 (1.106-1.741) and 1.700 (1.370-2.110), respectively. The rates on awareness, treatment and control in DM were 73.38%, 59.42% and 36.36% respectively. Among those who had knowledge on DM, the rates on treatment and on control were 80.97% and 61.20%. CONCLUSION: Our results showed that the prevalence of DM was high in the communities and it is essential to further increase the awareness, treatment and control rates on DM. PMID- 19565867 TI - [The relationship between emotional, physical abuse and Internet addiction disorder among middle school students]. AB - OBJECTIVE: To study the relationship between emotional, physical abuse and internet addiction disorder (IAD) among middle school students, in order to lay foundation for the development of prevention and control programs on IAD. METHODS: Students selected from 76 classes in Grade One and Grade Two, filled out the anonymous questionnaire, which including demographic characteristics of students, Young's Internet Addiction Scale, Parent-Child Conflict Tactics Scales, etc. Non-conditional multivariate logistic regression model was employed to explore the relationship between child abuse and IAD. Independent variable was child abuse while dependent variable was IAD. Covariant variables would include demographic characteristics, as sex, age, style of household, et al. RESULTS: Among 3798 students, the overall prevalence of emotional abuse was 81.1%, with 55.1% of them reported having experienced physical abuse. Among 3507 internet users, 5.2% was diagnosed as IAD. Results from the non-conditional multivariate logistic regression model showed that odds ratio of severe physical abuse and moderate physical abuse were 3.02 (1.63-5.58) and 4.00 (2.01-7.93). CONCLUSION: Moderate physical abuse and severe physical abuse were possible risk factors of IAD. PMID- 19565868 TI - [A case-control study on family environment related factors in attention deficit hyperactivity disorder with anxiety disorder]. AB - OBJECTIVE: To study the family rearing pattern of attention deficit hyperactivity disorder (ADHD) with or without anxiety disorder and to explore its risk factors. METHODS: 9495 children and their parents were sampled at random in Hunan province, using two-stage investigation. Those who were diagnosed ADHD and the normal control filled out Egna Minnen av Barndoms Uppfostran and family adaptability and cohesion scale by themselves. RESULTS: The comparison of factors as: actual family cohesion, parents' punishments, rejection, mother's excessive protection, intervention and father's excessive protection were significantly different between ADHD with or without anxiety disorder and normal children (P < 0.05). The comparison of parents' punishments, rejection, excessive protection and intervention were obviously different between ADHD with anxiety disorder and simple ADHD (P < 0.05). Mother's rejection was the influencing factor of simple ADHD, with OR as 1.122. Ideal family cohesion, mother's rejection and father's punishments were the influencing factors of ADHD with anxiety disorder, with OR as 0.966, 1.215 and 1.089 respectively. CONCLUSION: There were some problems in the parental rearing pattern of ADHD with or without anxiety disorder. Mother's rejection, father's punishments and ideal family cohesion were suggested to be correlated with ADHD and anxiety disorder. PMID- 19565869 TI - [Research on the structure equation model of tendency on drug abuse and related influencing factors in middle school students]. AB - OBJECTIVE: To understand the relationship of tendency on drug abuse and related influencing factors for middle school students. METHODS: A total of 603 students were assessed with self-made copying style scale, consciousness to resistance of smoking, drinking and resistance ability through a questionnaire. A relationship model was set up by structure equation modeling. RESULTS: Copying style and resistance ability had significantly direct influence on tendency of drug abuse (with standardized coefficients were 0.202 and 0.092 respectively). Consciousness to resistance of smoking and drinking had significantly indirect influence on tendency of drug abuse. The goodness of fit for the structural equation model was satisfactory, with GFI, AGFI, RMSEA as 0.981, 0.972 and 0.014 respectively. CONCLUSION: It was meaningful to grasp some copying skills and resistance ability to lower the tendency of drug abuse in middle school students. PMID- 19565870 TI - [HIV and syphilis infections among men who have sex with men in Chongqing municipality, China]. AB - OBJECTIVE: To examine the prevalence and predictors of HIV and syphilis infections among men who have sex with men (MSM) in Chongqing municipality, China. METHODS: Cross-sectional studies were conducted in three districts of Chongqing municipality from July to September, 2006 and 2007, respectively. Questionnaire-based interviews were conducted to provide information on demographic characteristics, behaviors on sex and drug use, and HIV and sexually transmitted diseases (STD) related health services. Blood samples were tested for HIV and syphilis infections. RESULTS: Of 1773 participants, 10.6% were HIV positive and 8.4% were sero-positive for syphilis infection. Factors independently associated with HIV infection included older age (26-35 year: AOR = 1.7, 95% CI: 1.1-2.6; >35 years of age: AOR = 2.3, 95% CI: 1.4-3.8), low level of education (AOR = 2.3, 95% CI: 1.5-3.5), having had multiple male sex partners (AOR = 1.8, 95% CI: 1.2-2.8), venues of recruitment from bathhouses/sauna (AOR = 2.3, 95% CI: 1.1-4.7), with self-reported STD symptoms in the past 12 months (AOR = 1.8, 95% CI: 1.2-2.7) and syphilis infection (AOR = 1.7, 95% CI: 1.1-2.9). CONCLUSION: The prevalence rates on both HIV and syphilis infections were considered to be high among MSM in Chongqing city. Unprotected anal sex and multiple sex partners were common in this group. Education, condom promotion, combination of HIV and STD screening, and treatment to the diseases are essential to this population in Chongqing. However, MSM had not hitherto been perceived to be in the epidemic mainstream in this area. PMID- 19565871 TI - [Factors influencing the rate on retention to methadone maintenance treatment program among heroin addicts in Guizhou, China]. AB - OBJECTIVE: To understand the situation of client-retention to methadone maintenance treatment (MMT) program and related factors. METHODS: A cohort study was adopted. In total, 1003 heroin addicts who were treated at 8 local MMT clinics with less than one month period, were recruited under nominal informed consent from Guizhou province, southwest part of China, during June to October 2006. Face-to-face interview and questionnaire administered to collect relevant information from the clients who were also followed until June 2007 to understand the situation on retention. Data were analyzed with Kaplan-Meier method to estimate the retention rate at different time spans while factors related to retention were analyzed with Cox proportional hazard regression model. RESULTS: All the clients were followed-up for 14 months, with an average retention of ten months. The retention rates of the clients were 68.8% and 57.4% at 6th-month and 12th-month of the treatment program, estimated by Kaplan-Meier method. Results of Cox regression analysis showed that factors influencing retention rate on MMT among the clients, including their awareness on MMT, daily dose of methadone intake, and different MMT clinics which they were attached to. Risk influencing the withdrawal from MMT had a 20% decrease along with the increase when the daily dose of methadone intake reached 25 mg, with a hazard ratio of 0.80 (P < 0.01). If the clients were aware that methadone was a life-time treatment when they began the MMT program, the risk for withdrawal would be lower than those who were not and the hazard ratio became 0.66 (P < 0.05). CONCLUSION: Our data showed that about half of the clients who were at the MMT program would still stick to it after 12-months, suggesting that the retention rate was not satisfactory and need to be improved. Factors as the MMT clinics themselves that the clients visited, daily methadone dosage they took, and the awareness on MMT etc. were important predictors to the rate of retention on MMT program. PMID- 19565872 TI - [Acceptability of vaginal microbicides among female sex workers in urban Beijing, China]. AB - OBJECTIVE: To explore the attitudes and willingness towards vaginal microbicides use among female sex workers (FSWs) for HIV/AIDS and sexually transmitted infection (STI) prevention in an urban district of Beijing, China, as well as its related factors. METHODS: A cross-sectional study using convenience sampling was conducted. A total of 54 FSWs were recruited from Shijingshan District in urban Beijing. Main outcome variables were measured by microbicide acceptability score with questionnaire, including high-risk sexual behaviors, self-reported condom use, self-reported HIV/STI history and perception on the risk of HIV/STI. RESULTS: Mean score of microbicide acceptability in FSWs was 2.73 (ranging 1 to 4). Acceptability score varied by the type of their sex partner (P = 0.049), experience of HIV testing (P = 0.037) and concerns about contracting STI (P = 0.042). Willingness to use and covert use of microbicides in FSWs varied significantly with their sex partners (P = 0.001). CONCLUSION: FSWs in Shijingshan district might have a positive response to vaginal microbicides use across all hypothetical characteristics in general. Further study is needed for comprehensive understanding of contextual factors related to it. PMID- 19565873 TI - [A seroepidemiologic analysis of hepatitis B in Sichuan province]. AB - OBJECTIVE: To understand the changes of hepatitis B infection rates, before and after the hepatitis B vaccine was included into EPI, and to evaluate the effect of immunization which would lead to the development of a more appropriate hepatitis B control strategy. METHODS: Seroepidemiologic method, with multi section random sampling method were chosen. 14 sites from 8 counties were involved. 2-4 ml of the vein blood was drawn from all the individuals engaged in the study including 3806 samples. HBsAg, anti-HBs, anti-HBc of the samples were tested with ELISA. RESULTS: Standardized positive rates of HBsAg and HBsAb were found as 7.05% and 29.77% respectively with the overall infection rate of HBV as 40.30%. The hepatitis B vaccine coverage of the children under 15 years was 70.73% and the positive rates for both HBsAg and anti-HBs were 2.62% and 56.68%, respectively. The coverage of hepatitis B vaccine among children under 3 years was 83.44% and the positive rates of both HBsAg and anti-HBs were 1.47% and 67.69% respectively, hepatitis B vaccine coverage of children under 3 years was 85.77%, with positive rates of HBsAg and anti-HBs as 1.78% and 75.44% respectively. CONCLUSION: Results from our study revealed that since the introduction of hepatitis B vaccination, the prevalence rates of HBsAg and HBV infection had an obvious decline, especially in children aged under 15 years of old, suggesting that some changes had occurred in the epidemic characteristics of hepatitis B in Sichuan. PMID- 19565874 TI - [A cross-sectional survey on hepatitis B among general population in areas along Yangzi River in Anhui province]. AB - OBJECTIVE: To describe the epidemic status of hepatitis B along the Yangzi River area, in Anhui province. METHODS: A cross-sectional seroepidemiological survey was conducted and the samples were collected by stratified cluster sampling. Serological biomarkers to hepatitis B virus were tested using enzyme-linked immunosorbent assay (ELISA) reagents. RESULTS: Among 2282 people, the prevalence rates of HBsAg, HBsAb, HBeAg, HBeAb, HBcAb and hepatitis B virus infection were 9.8%, 43.5%, 1.0%, 7.8%, 10.3% and 40.6% respectively. The prevalence rate of HBsAg among males was higher than that of females (P < 0.05). The vaccination rate was 24.7%, higher in urban than in rural areas. The vaccination rate was high in children younger than ten years old and in students. The prevalence rates of HBsAg and HBV among people who had received vaccines were lower than those who had not. CONCLUSION: The standardized prevalence rates of HBsAg (9.3%) was high in the area along the Yangzi River in Anhui province. The vaccinate rate was low in the country side. Expanded vaccinate which can obviously reduce the prevalence rate of HBsAg should be enhanced. PMID- 19565875 TI - [Molecular epidemiology of norovirus in outbreaks of gastroenteritis in Zhejiang from 2006 to 2007]. AB - OBJECTIVE: To study the molecular epidemiological characteristics of Norovirus gastroenteritis outbreaks in Zhejiang. METHODS: During January 2006 and December 2007, fecal specimens of patients collected from outbreaks of acute viral gastroenteritis were tested for Norovirus. Epidemiological data were also collected. Noroviruses were detected by a reverse transcription polymerase chain reaction (RT-PCR) and Real-time RT-PCR. Some positive samples were randomly selected and RT-PCR products were sequenced. Comparing to the nucleotide sequences of norovirus genotype I, II reference strains from GenBank, sequence analysis was undertaken based on partial sequence of RNA dependent RNA polymerase (RdRp) and capsid protein (VP1) gene. RESULTS: 5 outbreaks of viral gastroenteritis caused by Norovirus were reported. A total of 63 stools were obtained from cases with acute gastroenteritis. Noroviruses alone were detected in 45 cases and the illness appeared in autumn. Phylogenetic analysis revealed that Norovirus belonged to G II/G II 4 type. The strains isolated from Zhejiang were almost identical on G II/4 variants that causing epidemics in Beijing and in The Netherlands with the homology of 99.7% and 98.5%-99.0% respectively. Phylogenetic analysis revealed that the isolates were located at the same branch as the norovirus G II/4 variants found in Beijing and Netherlands. CONCLUSION: Norovirus is a major cause of outbreaks of viral gastroenteritis in Zhejiang province Genogroup II/4 variants viruses were the prevalent strains. PMID- 19565876 TI - [Study on the relationship between familial clustering of hepatocellular carcinoma and polymorphism of cytochrome P450 2E1 gene in Zhuang population, Guangxi]. AB - OBJECTIVE: To study the relationship between familial clustering of hepatocellular carcinoma (HCC) and the polymorphism of cytochrome P450 2El gene (CYP2E1) as well as of other relevant risk factors to the cancer. METHODS: Peripheral blood samples were collected from 91 members of 10 HCC clustering families and 102 of 10 control families, among Zhuang population, in Guangxi. The area had been with high incidence rate of HCC. Genotypes and allele frequencies of CYP2E1 Rsa I site were determined by polymerase chain reaction, combined with restriction fragment length polymorphism method (PCR-RFLP). Serum HBsAg was tested by means of ELISA. Data on relevant risk factors of the cancer were collected as well, through a unique questionnaire. RESULTS: Frequencies of c1/c1 and c1/c2 genetypes of CYP2E1 Rsa I site were 63.7% and 36.3%, respectively, in the members of families with cancer clustering phenomena. In the members of the control families, these two rates were 48.0% and 52.0%, respectively (OR = 1.901, 95% CI: 1.067-3.387). Difference of genotypes frequencies of CYP2E1 Rsa I site between the members in these two groups was statistically significant (chi2 = 4.797, P = 0.029). According to the results from non-condition logistic regression analysis, the major risk factors on familial clustering of HCC could be listed as: intake of corns, HBsAg carrying status and CYP2E1 c1/c1 genotype. CONCLUSION: The relationship seemed to exist between familial clustering of HCC and the frequencies of polymorphism of cytochrome P450 2E1 gene (CYP2E1). The frequencies of CYP2E1 Rsa I site were neither the only nor the major factor, causing the familial clustering phenomenon of cancer. More possible, it was the affect of syntheses with the involvement of multiple factors. PMID- 19565877 TI - [Survey on thalassemia among people of reproductive age in Guilin City, Guangxi, China]. AB - OBJECTIVE: To study the present situation of thalassemia among people at reproductive age in Guilin city. METHODS: A complete red blood cell mean cell volume (RBCMCV) was detected as well as hemoglobin electrophoresis analysis were done for all samples. Suspected alpha or beta thalassemia cases, were screened out and alpha and beta thalassemia genes were detected by PCR-RDB. RESULTS: Among 1580 cases, 79 cases were detected a thalassemia gene positive, with the detection rate as 5.00%. The detection rates on a thalassemia were 5.32%, 4.68% in males and females, but no significant difference (chi2 = 3.04, chi2 < chi0.05 (1) = 3.84, P > 0.05). 114 cases were detected carrying beta thalassemia gene with the detection rate as 7.22%. The detection rates on beta thalassemia were 7.85%, 6.58% in males and females, with no significant difference (chi2 = 0.95, chi2 < chi0.05(l) = 3.84, P > 0.05). The overall detection rate of thalassemia was 12.22% (193/1580). Alpha thalassemia were found to have had ten genotypes, with --SEA/alphaalpha the most common one and the detection rate was 3.54%. Alpha(3.7)/alphaalpha appeared the majority in the static alpha thalassemia and - SEA/alphaalpha took the majority in the light a thalassemia, while --SEA-alphaCS was the major one in the intermedia alpha thalassemia, with gene contributions of 7.59%, 70.88% and 2.53%. Beta thalassemia was detected having seven genotypes,with CD41-42 (-TTCT) the most common one in beta thalassemia, and the detection rate was 3.16%. The commonly seen three mutations, CD41-42 (-TTCT), CD17 (A-->T) and IVS-II-654 (C-->T) were accounted for 87.71% of beta thalassemia. The detection rate on thalassemia a and beta combination was 0.63%. CONCLUSION: The detection rate of thalassemia among people at productive age in Guilin city was relatively high. PMID- 19565878 TI - [Comparison between early outbreak detection models and simulated outbreaks of measles in Beijing]. AB - OBJECTIVE: Using simulated outbreaks to choose the optimal model and its related parameters on measles so as to provide technical support for developing an Auto Warning System (AWS). METHODS: AEGIS-Cluster Creation Tool was applied to simulate a range of unique outbreak signals. Then these simulations were added to the actual daily counts of measles from the National Disease Surveillance System, between 2005 and 2007. Exponential weighted moving average (EWMA), C1-MILD (C1), C2-MEDIUM (C2), C3-ULTRA (C3) and space-time permutation scan statistic model were comprehensively applied to detect these simulations. Tools for evaluation as Youden' s index and detection time were calculated to optimize parameters before an optimal model was finally chosen. RESULTS: EWMA (lamda = 0.6, k = 1.0), CI (k = 0.1, H=3sigma), C2 (k = 0.1, H=3sigma), C3 (k = 1.0, H=4sigma) and space-time permutation scan statistic (maximum temporal cluster size=7 d, maximum spatial cluster size = 5 km) appeared to be the optimal parameters among these models. Youden's index of EWMA was 90.8% and detection time being 0.121 d. Youden's index of C1 was 88.7% and detection time being 0.142 d. Youden's index of C2 was 92.9% and detection time being 0.121 d. Youden's index of C3 was 87.9% and detection time being 0.058 d. Youden's index of space-time permutation scan statistic was 94.3% and detection time being 0.176 d. CONCLUSION: Among these five early warning detection models, space-time permutation scan statistic model had the highest efficacy. PMID- 19565880 TI - [Using spatial autocorrelation analysis to study spatial heterogeneity of liver cancer in Guangxi]. AB - OBJECTIVE: To study the spatial distribution characteristics of liver cancer in Guangxi so as to provide evidence for the development of control and prevention on liver cancer. METHODS: The average eight year morbidity was computed, using the rates of liver cancer in 2000-2007. The spatial statistics module of GIS was used to conduct spatial autocorrelation analysis, and the disease mapping was drawn, using the Map Info 8.0 software. RESULTS: The average morbidity rate was clustered in Guangxi in the past eight years, with Moran's I index as 0.34 and P value below 0.01. G index appeared to be 0.77 and the P value was below 0.01. Moran's I correlogram lifted up in four spaces, specifically, the cluster took place in both macro-scale (one to three spatial intervals, 45 to 135 km real scale) and micro-scale (16 to 18 spatial intervals, 720 to 800 km real scale). When the spatial interval became 14 and real scale was 60 km, the spatial distribution of liver cancer showed the most intensive autocorrelation. Most of the regions with high morbidity would be clustered in the southwest and southern parts, along the coastal areas of Guangxi while the regions with low morbidity clustered in the northern part of Guangxi. CONCLUSION: Liver cancer was found un randomly distributed and geographically clustered in Guangxi in 2000-2007. PMID- 19565879 TI - [Using two-dimensional graphic cluster method to study the geographic distribution pattern of patients with anorectal atresia/stenosis, in China]. AB - OBJECTIVE: To investigate the geographic distribution pattern of patients with anorectal atresia/stenosis in China, in order to provide clue for research on its etiology. METHODS: Data were collected from Chinese Birth Defects Monitoring Network (CBDMN), which was a hospital-based congenital malformations registry system. From 2001 to 2005, all fetuses with more than 28 weeks of gestation and neonates up to 7 days of age, were monitored. Two-dimensional graphic cluster method was used to divide monitoring stations into different classes with the incidence rates of anorectal atresia/stenosis. RESULTS: The overall incidence of anorectal atresia/stenosis was 3.17 per 10,000 during 2001 to 2005. The incidence was higher in Eastern than that in Mid or Western parts of China and the difference was statistically significant (z = 2.50, 3.69; P = 0.012, < 0.001). The monitoring stations were grouped into 6 classes. Class I was with Heilongjiang, Jilin and Liaoling; Class II was with Fujian, Guangdong, Hainan, Guangxi, and South Hunan and Jiangxi; Class III was with Beijing, Tianjin, Hebei, Shandong, and North Jiangsu and Anhui; Class IV was with Zhejiang, Shanghai, South Anhui and Jiangsu, North Hunan and Jiangxi, Hubei, Henan, Shanxi and Inner Mongolia, Class V was with Ningxia, Gansu and Qinghai; and Class VI was with Shaanxi, Sichuan, Chongqing, Yunnan, Guizhou, Xinjiang and Tibet. CONCLUSION: Our findings discovered the geographic distribution patterns of patients with anorectal atresia/stenosis in China. It is important to further analyze the relevant environmental factors attached to it so a better regional monitoring system for anorectal atresia/stenosis can be operated. PMID- 19565881 TI - [Cloning and expression of the nucleoprotein gene of Puumala-like virus]. AB - OBJECTIVE: In order to detect Hokkaido virus (HOKV), a recombinant baculovirus containing the nucleoprotein (NP) gene of HOKV was constructed, and then the NP was expressed in insect cell. METHODS: The NP gene was cloned into plasmid PCR 2.1TA vector and then was ligated into baculovirus donor plasmid pFastBac after cutting by the restriction enzyme Kpn I and Not I. pFastBac 1 was subsequently transferred into the One Short TOP10 competent cells and then into DH1OBac E. coli competent cells, which contained the baculovirus shuttle vector (Bacmid) and the helper plasmid to generate a recombinant bacmid. RESULTS: The NP gene was successfully expressed in Sf9 insect cell. The expressed recombinant nucleoprotein had been identified in the Sf9 insect cell by indirect immunofluorescence assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. The results showed that the recombinant nucleoprotein appeared a molecular weight of 50 x 10(3) Mr, and could reacted with anti-recombinant Puumala virus (PUUV) nucleocapsid monoclonal antibodies and polyclonal antibodies against hantavirus. CONCLUSION: Our results indicated that the recombinant nucleoprotein was successfully expressed and having the immunogenicity and reactivity of natural nucleoprotein of HOKV. PMID- 19565882 TI - [Study on the difference of genes and the type identification of hantavirus from Lishui, Zhejiang province]. AB - OBJECTIVE: To isolate hantavirus from Lishui county--one of the epidemic regions for hemorrhagic fever with renal syndrome (HFRS), in Zhejiang province, and to identify the serotype and molecular/biological characteristics of a new HTN subtype hantavirus (HV) strains, hopefully to provide evidence for HFRS prevention and therapy. METHODS: Data on the host animals was collected from Lishui, Zhejiang province in 2007. Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues from HV infected Vero-E6 cells for HV isolation, then total RNA was extracted from Hantavirus Lishui strains and amplified by RT-PCR M, S segments of strains genome were also cloned and sequenced and compared with those of other strains of HV. RESULTS: 2 strains virus (ZLS6-11 and ZLS-12) were successfully isolated from 7 positive lung samples of mice and were identified as HTNV by anti-McAb and phylogenetic analysis. With sequence compation,we found that 2 strains with complete M and S segment had higher homology with HTN-type strains than with other types of HV, but 13.4%-20.7% and 10.3%-16.1% of the genes were found which were different from HTNV. The phylogenetic trees constructed by complete S and M segment showed that ZLS6-11 and ZLS-12 strains were located in HTNV group,and structured independent embranchment. CONCLUSION: ZLS6-11 and ZLS-12 Strains were believed to belong to HTN-type and phylogenetically different from the HTNV. PMID- 19565884 TI - [Prognostic value of right ventricular dysfunction and derivation of a prognostic model for patients with acute pulmonary thromboembolism]. AB - OBJECTIVE: Acute pulmonary thromboembolism (PTE) patients with right ventricular dysfunction (RVD) may benefit from thrombolytic therapy but may end up with worse prognosis. RVD was assessed in prognosis to which a model on it was constructed to decide the indexes correlated to the best prognosis. METHODS: This prospective study included 520 consecutive acute PTE patients from 41 hospitals in China between June 2002 and February 2005. All the patients were evaluated by transthoracic echocardiography (TTE), CT pulmonary angiography (CTPA), laboratory tests, and blood gas analysis. Physicians were asked to record all the clinical manifestations. Data from Univariate analysis demonstrated the parameters correlated with an 14-day clinically adverse outcomes. Multiple logistic regression analysis was used to decide the independent predictors and to construct a prognostic model. RESULTS: Mean age of the patients was 57.4 +/- 14.1 years and 323 of them (62.1%) were male. The 14-day mortality in normotensive patients with RVD was higher' (2.0% vs. 0.4%, P < 0.01) than those without. ROC curve showed the best cut-off values of RVED/LVED and SPAP for 14-day prognosis were 0.67 and 60 mm Hg, respectively. SPAP independently predicted 3-month clinical outcomes (P < 0.01). Results from Univariate analysis demonstrated that 24 parameters were correlated with an adverse 14-day clinical outcomes, which include palpation, syncope, panic, cyanosis, respiratory rate > or = 30/min, pulse > or = 110/min, jugular vein, accentuation of P2, murmurs in tricuspid area, time interval from onset, creatine phosphokinase (CPK), lactate dehydrogenase (LDH), alveolar-arterial PO2 difference (PA-aO2), white blood cell (WBC) < 4.0 x 10(9)/L or >10.0 x 10(9)/L, platelet, thrombus on TTE, RV/LV > 1.1, TRPG >30 mm Hg, IVCmin <8 mm, RVD, RVED/LVED > 0.6, SPAP >60 mm Hg, RVWM, PTE range larger than two lobes or seven segments on CTPA. Furthermore, a multiple logistic regression model implied 8 predictors including RVD, RVED/LVED > 0.6, SPAP >60 mm Hg, pulse > or = 110/min, accentuation of P2, Syncope, CPK, WBC < 4.0 x 109/L or > 10.0 x 10(9)/L be independent predictors of an 14-day clinically adverse outcome (P < 0.01). This model seemed to fit well (P < 0.001). We chose a cut-off value as P > or = 0.2 and compared the model to the original derivation samples. Data showed that the sensitivity (true positive rate) was 81.82%, specificity was 92.11%, false positive rate was 18.18%, coincidence was 91.14%, and the concordance rate was 80.96%. CONCLUSION: RVD seemed a nice discriminator for poor prognosis in normotensive patients. Early detection of RVD (especially RVED/LVED > 0.67 and/or SPAP >60 mm Hg) was beneficial for identifying patients at high-risk and the multiple logistic regression model (P < 0.001) could be well fitted. PMID- 19565883 TI - [Study on the efficacy and safety of short-term treatment including fluoroquinolones anti-tuberculosis drugs for rifampicin resistant pulmonary tuberculosis]. AB - OBJECTIVE: To evaluate the efficacy and safety of short-term treatment including fluoroquinolones anti-tuberculosis drugs for rifampicin resistant pulmonary tuberculosis (TB) in those areas carrying out the 'TB control project'. METHODS: TB cases involved in this study were from TB drug resistance surveillance in Heilongjiang province, Zhejiang province and Shenzhen city from 2004 to 2006. TB cases with rifampicin resistant were randomly divided into the treatment group (including fluoroquinolones anti-tuberculosis drugs group) and the control group (re-treatment regimen group). The treatment group was treated with 3RFT AM Ofx Pto PAS-INH/5RFT Ofx Pto PAS-INH while the control group was treated with 3 H3R3Z3E3S3/5 H3R3E3. Efficacy of short-term treatment was analyzed by per protocol analysis (PP analysis) and intention-to-treat analysis (ITT analysis) while drug adverse reactions was also observed. RESULTS: (1) 154 patients with rifampicin resistant pulmonary tuberculosis were recruited among them, 25 (16.2%) were only resistant to rifampicin, 114 (74.0%) to MDR-TB and 15 (9.8%) to others (resistant R+S, resistant R+E and resistant R+E+S). 114 TB cases completed the full course of treatment,with 71 in the treatment group and 43 in the control group. (2) Sputum negative conversion rate of the treatment group and the control group were 78.9% and 65.1% (chi2CMH = 4.558, P = 0.011) respectively, by per protocol analysis. Sputum negative conversion rate of the treatment group and the control group were 65.9% and 40.6% (chi2CMH = 0.272, P = 0.001) respectively, by intention-to-treat analysis. The sputum negative conversion rate of the treatment group was higher than in the control group when treating rifampicin resistant pulmonary tuberculosis and MDR-TB patients. (3) Three patients withdrew in each of the two groups because of adverse effects to the drugs. Rates of adverse reaction to drugs appeared to be 23.9% (17/71) and 18.6% (8/43) in the treatment and in the control groups, with no statistically significant difference between the two groups. CONCLUSION: The efficacy of treatment including fluoroquinolones anti-tuberculosis drugs group seemed better than the re-treatment regimen group in treating patients with rifampicin resistant pulmonary tuberculosis and those MDR-TB patients. PMID- 19565885 TI - [A case-control study on the association of RFC-1 polymorphism and cervical cancer]. AB - OBJECTIVE: To evaluate the possible association between RFC-1 polymorphism and cervix carcinoma, as well as the interaction between polymorphism and human papilloma virus16 (HPV16). METHODS: Based on a hospital-based case-control study, 107 cases which were diagnosed as cervical cancer pathematologically and 107 controls with hysteromyoma, were selected by frequency, matched with age and habitation. HPV16 and RFC-1 A80G polymorphism were detected by special PCR and RFLP. RESULTS: (1) HPV16 infection rate in cases (56.07%) was higher than that in controls (31.78%) with the adjusted OR of 3.25 (95% CI: 1.74-6.08). (2) All the genotypes were tested by Hardy-Weinberg balance. (3) Compared with RFC-1 AA,RFC-1 GG had higher risk for cervical cancer with OR of 2.42 (95% CI: 1.01-5.81). (4) No statistical significance was noticed regarding the interaction between RFC-1 polymorphism and HPV16 in logistic regression method. CONCLUSION: The introduction of RFC-1 80GG gene type could increase the risk of cervical cancer. PMID- 19565886 TI - [Progress on studies of genotype and prevalence of hepatitis E virus infection in animals]. PMID- 19565887 TI - [The acceptability on vaginal microbicides to prevent human immunodeficiency virus/sexually transmitted diseases]. PMID- 19565888 TI - [Surveillance on human immunodeficiency virus incidence in the United States]. PMID- 19565889 TI - [A multicentre study of Shigella spp in Henan province, during 2000-2007]. PMID- 19565890 TI - [Epidemiological investigation on an outbreak of leptospirosis involving 36 patients]. PMID- 19565891 TI - [Correlates of and willingness to participate in HIV routine counseling and testing among female sex workers in a city of Yunnan province]. PMID- 19565892 TI - [Study on the change of quality of life and its influential factors for drug abusers accepting methadone maintenance treatment]. PMID- 19565893 TI - [A study of mix-infections with different genotypes of hepatitis C virus in patients from a methadone maintenance clinic in Wuhan]. PMID- 19565894 TI - The development of gene therapy: from monogenic recessive disorders to complex diseases such as cancer. AB - During the last 4 decades, gene therapy has moved from preclinical to clinical studies for many diseases ranging from monogenic recessive disorders such as hemophilia to more complex diseases such as cancer, cardiovascular disorders, and human immunodeficiency virus (HIV). To date, more than 1,340 gene therapy clinical trials have been completed, are ongoing, or have been approved in 28 countries, using more than 100 genes. Most of those clinical trials (66.5%) were aimed at the treatment of cancer. Early hype, failures, and tragic events have now largely been replaced by the necessary stepwise progress needed to realize clinical benefits. We now understand better the strengths and weaknesses of various gene transfer vectors; this facilitates the choice of appropriate vectors for individual diseases. Continuous advances in our understanding of tumor biology have allowed the development of elegant, more efficient, and less toxic treatment strategies. In this introductory chapter, we review the history of gene therapy since the early 1960s and present in detail two major recurring themes in gene therapy: (1) the development of vector and delivery systems and (2) the design of strategies to fight or cure particular diseases. The field of cancer gene therapy experienced an "awkward adolescence." Although this field has certainly not yet reached maturity, it still holds the potential of alleviating the suffering of many individuals with cancer. PMID- 19565895 TI - Designing adenoviral vectors for tumor-specific targeting. AB - Adenovirus provides an attractive candidate tool to destroy tumor cells. However, to fulfill the expectations, selective targeting of tumor cells is mandatory. This chapter reviews critical aspects in the design of tumor-targeted adenovirus vectors and oncolytic adenoviruses. The review focuses on genetic modifications of capsid and regulatory genes that can enhance the therapeutic index of these agents after systemic administration. Selectivity will be considered at different levels: biodistribution selectivity of the injected virus particles, transductional selectivity defined as cell receptor interactions and trafficking that lead to virus gene expression, transcriptional selectivity by means of tumor selective promoters, and mutation-rescue selectivity to achieve selective replication. Proper assays to analyze selectivity at these different levels are discussed. Finally, mutations and transgenes that can enhance the potency and efficacy of tumor-targeted adenoviruses from virocentric or immunocentric points of view will be presented. PMID- 19565896 TI - Analysis of HSV oncolytic virotherapy in organotypic cultures. AB - Tumor-selective replication-competent viral vectors, such as oncolytic herpes simplex virus (HSV) type I (HSV-1), represent an attractive strategy for tumor based therapies because these viruses can replicate and spread in situ exhibiting cytopathic effects through direct oncolytic activity. These lytic viruses offer a distinct advantage over other forms of cancer therapies in that they are self perpetuating and can spread not only in the tumor itself, but also to distant micrometastases. Translational studies aimed at identifying novel virotherapies for human cancers are incumbent upon the appropriate experimental models. While animal models are the preferred choice for efficacy studies of HSV virotherapy, we have developed a novel complementary approach toward assessing the effectiveness of oncolytic HSV therapy in both brain and prostate cancers. This experimental model takes advantage of previously published work in which human prostate cancer biopsies and rodent brain slices can be easily maintained ex vivo. The advantage of these systems is that the three-dimensional structure remains intact. Thus, all of the factors that may affect viral entry and replication, such as cell-cell and cell-matrix interactions, and interstitial fluid within this three-dimensional milieu remain preserved. Moreover, with respect to the brain, this system offers the advantage of direct access to brain cells, such as microglia and astrocytes, and circumvents the problems associated with the presence of the blood-brain barrier. PMID- 19565897 TI - Use of minicircle plasmids for gene therapy. AB - A large number of cancer gene therapy clinical trials are currently being performed that are attempting to evaluate novel approaches to eliminate tumor cells by the introduction of genetic material into patients. One of the most important objectives in gene therapy is the development of highly safe and efficient vector systems for gene transfer in eukaryotic cells. Currently, viral and nonviral vector systems are used, both having their advantages and limitations. Minicircles are novel supercoiled minimal expression cassettes, derived from conventional plasmid DNA by site-specific recombination in vivo in Escherichia coli for the use in nonviral gene therapy and vaccination. Minicircle DNA lacks the bacterial backbone sequence consisting of an antibiotic resistance gene, an origin of replication, and inflammatory sequences intrinsic to bacterial DNA. In addition to their improved safety profile, minicircles have been shown to greatly increase the efficiency oftransgene expression in various in vitro and in vivo studies. In this chapter, we describe the production, purification, and application of minicircle DNA and discuss the rationale of the improved gene transfer efficiencies compared to conventional plasmid DNA. PMID- 19565898 TI - Transposable elements as plasmid-based vectors for long-term gene transfer into tumors. AB - A primary limitation to using nonviral vectors for cancer gene therapy is transient expression of the therapeutic gene. Even when the ultimate goal is tumor cell death, a minimum threshold of gene expression is required to kill tumor cells by direct or indirect mechanisms. It has been shown that transposable elements can significantly enhance the duration of gene expression when plasmid DNA vectors are used to transfect tumor or tumor-associated stroma. Much like a retrovirus, transposon-based plasmid vectors achieve integration into the genome, and thereby sustain transgene expression, which is especially important in actively mitotic cells such as tumor cells. Herein we briefly discuss the different transposons available for gene therapy applications, and provide a detailed protocol for nonviral transposon-based gene delivery to solid experimental tumors in mice. PMID- 19565899 TI - Designing plasmid vectors. AB - Nonviral gene therapy vectors are commonly based on recombinant bacterial plasmids or their derivatives. The plasmids are propagated in bacteria, so, in addition to their therapeutic cargo, they necessarily contain a bacterial replication origin and a selection marker, usually a gene conferring antibiotic resistance. Structural and maintenance plasmid stability in bacteria is required for the plasmid DNA production and can be achieved by carefully choosing a combination of the therapeutic DNA sequences, replication origin, selection marker, and bacterial strain. The use of appropriate promoters, other regulatory elements, and mammalian maintenance devices ensures that the therapeutic gene or genes are adequately expressed in target human cells. Optimal immune response to the plasmid vectors can be modulated via inclusion or exclusion of DNA sequences containing immunostimulatory CpG sequence motifs. DNA fragments facilitating construction of plasmid vectors should also be considered for inclusion in the design of plasmid vectors. Techniques relying on site-specific or homologous recombination are preferred for construction of large plasmids (>15 kb), while digestion of DNA by restriction enzymes with subsequent ligation of the resulting DNA fragments continues to be the mainstream approach for generation of small- and medium-size plasmids. Rapid selection of a desired recombinant plasmid against a background of other plasmids continues to be a challenge. In this chapter, the emphasis is placed on efficient and flexible versions of DNA cloning protocols using selection of recombinant plasmids by restriction endonucleases directly in the ligation mixture. PMID- 19565900 TI - Development of bacterial vectors for tumor-targeted gene therapy. AB - Gene therapy holds great promise for the treatment of cancer. The success of the strategy relies on effective gene transfer into tumor microenvironments. Although a variety of gene delivery vehicles, such as viral vectors, has been developed, most of them suffer from some limitations, including inadequate tumor targeting, inefficient gene transfer, and potential toxicity. This situation suggests that it is necessary to develop novel vectors for effective tumor-targeted gene transfer. The discovery of tumor-targeting bacteria has spurred interest in the use of these bacteria as gene transfer vectors. In this review, we focus on the current status of the development of bacterial vectors for cancer gene therapy and highlight some of the directions that the field may take. PMID- 19565901 TI - Electroporative gene transfer. AB - Membrane electroporation (MEP) uses short high-voltage pulses to render cell membranes transiently porous and therewith permeable to otherwise impermeable substances. This technique was first described, in vitro, by Neumann in 1982. In vivo, this method is restricted to solid tissues accessible to the electrodes used to apply the electric field pulses. Electroporation of cell tissue gains increasing importance especially in clinical applications such as electrochemotherapy (ECT) of, e.g., skin tumors, and for gene therapy. The various applications of MEP include, in addition to the direct functional transfer of genes (electrotransfection, electrogenetransfer) and drugs, the release of proteins, and the electrotransfer of ionic dyes into cells. But, nevertheless, the mechanism of pore opening and resealing as well as the transfer, especially of DNA, is not yet completely understood. PMID- 19565902 TI - Gene gun delivery systems for cancer vaccine approaches. AB - Gene-based immunization with transgenic DNA vectors expressing tumor-associated antigens (TAA), cytokines, or chemokines, alone or in combination, provides an attractive approach to increase the cytotoxic T cell immunity against various cancer diseases. With this consideration, particle-mediated or gene gun technology has been developed as a nonviral method for gene transfer into various mammalian tissues. It has been shown to induce both humoral and cell-mediated immune responses in both small and large experimental animals. A broad range of somatic cell types, including primary cultures and established cell lines, has been successfully transfected ex vivo or in vitro by gene gun technology, either as suspension or adherent cultures. Here, we show that protocols and techniques for use in gene gun-mediated transgene delivery system for skin vaccination against melanoma using tumor-associated antigen (TAA) human gpl00 and reporter gene assays as experimental systems. PMID- 19565903 TI - Ultrasound-mediated gene transfection. AB - Ultrasound-mediated gene transfection (sonotransfection) has been shown to be a promising physical method for gene therapy, especially for cancer gene therapy. The procedure being done in vitro uses several ultrasound exposure (sonication) setups. Although high transfection rates have been attained in some of these setups in vitro, replicating similar levels of transfection in vivo has been difficult. In vivo-simulated setups offer hope for a more consistent outcome in vivo. Presented in this chapter are typical methods of sonotransfection in vitro, methods when using a novel in vivo-simulated in vitro sonication setup and also sonotransfection methods when doing in vivo experiments. Factors that could potentially influence the outcome of an ultrasound experiment are cited. Several advantages of sonotransfection are recognized, although a low transfection rate is still considered a disadvantage of this method. To improve the transfection rate and the efficiency of sonotransfection, several studies are currently being undertaken. Particularly promising are studies using engineered microbubbles to carry the therapeutic genes into a particular target tissue in the body, then using ultrasound to release or deliver the genes directly into target cells, e.g., cancer cells. PMID- 19565904 TI - Nonviral jet-injection technology for intratumoral in vivo gene transfer of naked DNA. AB - The main challenges for application of gene therapy to patients are poor selectivity in vector targeting, insufficient gene transfer, and great difficulties in systemic treatment in association with safety concerns for particular vector systems. For success in gene therapy, safe, applicable, and efficient transfer technologies are required. Because of the complex nature of targeted vector delivery to the tumor, our strategy for gene therapy is focused on the development of local nonviral gene transfer. This approach of local interference with tumor growth and progression could contribute to better control of the disease. Transfer of naked DNA is an important alternative to liposomal or viral systems. Different physical procedures are used for improved delivery of naked DNA into the target cells or tissues in vitro and in vivo. Among the various nonviral gene delivery technologies, jet-injection is gaining increased attractiveness, because this technique allows gene transfer into different tissues with deep penetration of naked DNA by circumventing the disadvantages associated with, e.g., viral vectors. The jet-injection technology is based on jets of high velocity for penetration of the skin and underlaying tissues, associated with efficient transfection of the affected area. The jet-injection technology has been successfully applied for in vivo gene transfer in different tumor models. More importantly, the efficacy and safety of jet-injection gene transfer have recently been investigated in a phase I clinical trial. PMID- 19565905 TI - Methods for constructing and evaluating antitumor DNA vaccines. AB - An antitumor DNA vaccine is a bacterial DNA plasmid that encodes the complementary DNA (cDNA) of a tumor antigen. When injected into recipients, antitumor DNA vaccines have been shown to elicit both humoral and cellular immunity against the encoded tumor antigen. These vaccines represent a relatively new immunotherapeutic technique being investigated as a means to deliver a target antigen and elicit or augment antitumor antigen-specific immune responses. One of the primary advantages of DNA vaccines as opposed to some other methods of antigen delivery is that they can be easily constructed, purified, and delivered to recipients. In this review we describe this process, detailing the procedures used to construct, purify, deliver, and evaluate the efficacy of DNA vaccines. We begin by describing the process of molecularly constructing the vaccine, from selecting a bacterial plasmid to form the backbone of the vaccine, cloning the antigen cDNA into this plasmid, and confirming the sequence and orientation of the completed vaccine. This is then followed by a series of experiments that can be used to ensure that the antigen encoded by the vaccine is transcribed and translated after being taken up by eukaryotic cells. We then describe large-scale purification procedures that can be used to obtain sufficient quantities of plasmid DNA to conduct in vivo immunization experiments. Finally, we provide an immunization protocol that can be used to evaluate the immunological efficacy of the constructed DNA vaccine. By following these protocols, it is possible to construct, purify, deliver, and evaluate the efficacy of antitumor DNA vaccines. PMID- 19565906 TI - Immunity of lentiviral vector-modified dendritic cells. AB - Innovative approaches to induce a strong immune response are key to the success of immunotherapy. Dendritic cells (DCs) are professional antigen-presenting cells (APCs) equipped with co-stimulatory, adhesion, and major histocompatibility complex (MHC) molecules needed for initiation and reactivation of the immune response. DCs are able to initiate and stimulate both innate and adaptive immune responses and, by secretion of cytokines, chemokines, and expression of regulatory molecules, to shape the adaptive immune response toward a long-lasting memory immunity. DCs from the peripheral blood of immune-compromised patients, however, often display an immature phenotype with defective functions. This emphasizes the importance and potential of engineering antigen-specific DCs in vitro. A state-of-the-art approach to overcome the prevailing immune dysfunction(s) in patients is to engineer DCs or DC progenitors to generate fully functional DCs for the modification of host immunity. Lentiviral vectors (LVs) are highly efficient gene transfer vehicles for engineering DC functions. Examples oflentiviral vectors encoding immune-modulatory genes and useful functional assays for the analysis of effector immune cell response are described in this chapter. PMID- 19565907 TI - Saporin suicide gene therapy. AB - New genes useful in suicide gene therapy are those encoding toxins such as plant ribosome-inactivating proteins (RIPs), which can irreversibly block protein synthesis, triggering apoptotic cell death. Plasmids expressing a cytosolic saporin (SAP) gene from common soapwort (Saponaria officinalis) are generated by placing the region encoding the mature plant toxin under the control of strong viral promoters and may be placed under tumor-specific promoters. The ability of the resulting constructs to inhibit protein synthesis is tested in cultured tumor cells co-transfected with a luciferase reporter gene. SAP expression driven by the cytomegalovirus (CMV) promoter (pCI-SAP) demonstrates that only 10 ng ofplasmid DNA per 1.6 x 10(4) B16 melanoma cells drastically reduces luciferase reporter activity to 18% of that in control cells (1). Direct intratumoral injections are performed in an aggressive melanoma model. B16 melanoma-bearing mice injected with pCI-SAP complexed with lipofectamine or N-(2,3-dioleoyloxy-1 propyl) trimethylammonium methyl sulfate (DOTAP) show a noteworthy attenuation in tumor growth, and this effect is significantly augmented by repeated administrations of the DNA complexes. Here, we describe in detail this cost effective and safe suicide gene approach. PMID- 19565908 TI - Using in vivo biopanning for the development of radiation-guided drug delivery systems. AB - This chapter illustrates our protocol for in vivo biopanning using T7 bacteriophage libraries for the purpose of selecting recombinant peptides for the tumor-specific delivery of radiosensitizers to radiation-inducible antigens within tumor neovasculature. Our goal is to discover peptides binding within tumor vascular endothelium of irradiated tumors. We have previously demonstrated that tumor irradiation increases the spectrum of antigenic targets for drug delivery. To identify candidate peptides with the ability to bind radiation induced antigens, we inject the phage peptide library intravenously into mice bearing irradiated GL261 and Lewis lung carcinoma (LLC) hind limb tumors. Phage are recovered from excised tumors, amplified, and readministered to mouse-bearing tumors for six total rounds. At least 50 bacterial colonies are selected from each of the tumor types, and prioritized. This prioritization is based on their relative concentrations in tumor versus normal tissues, and then assessment of dominant phage present in both tumor types. These phage are amplified, and the gene sequences determined to deduce the recombinant peptide product. Further prioritization is performed by fluorescence labeling of the selected phage, and injection into irradiated and mock-irradiated tumor-bearing mice for evaluation of in vivo targeting of the candidate phage/peptides. PMID- 19565909 TI - Chemosensitization of tumor cells: inactivation of nuclear factor-kappa B associated with chemosensitivity in melanoma cells after combination treatment with E2F-1 and doxorubicin. AB - Combination chemotherapy has been shown to be more effective than single-agent therapy for many types of cancer, but both are known to induce drug resistance in cancer cells. Two major culprits in the development of this drug resistance are nuclear factor-kappaB (NF-kappaB) and the multidrug resistance (MDR) gene. For this reason, chemogene therapy is emerging as a viable alternative to conventional chemotherapy combinations. We have shown that transduction of the E2F-1 gene in melanoma cells markedly increases cell sensitivity to doxorubicin, thereby producing a synergistic effect on melanoma cell apoptosis. Our microarray results show that the NF-kappaB pathway and related genes undergo significant changes after the combined treatment of E2F-1 and doxorubicin. In fact, inactivation of NF-kappaB is associated with melanoma cell apoptosis induced by E2F-1 and doxorubicin, providing a link between the NF-kappaB signaling pathway and the chemosensitivity of melanoma cells after this treatment. PMID- 19565910 TI - Induction of tumor cell apoptosis by TRAIL gene therapy. AB - Members of the tumor necrosis factor (TNF) superfamily influence a variety of immunological functions, including cellular activation, proliferation, and death, upon interaction with a corresponding superfamily of receptors. Whereas interest in the apoptosis-inducing molecules TNF and Fas ligand has peaked because of their participation in events such as autoimmune disorders, activation-induced cell death, immune privilege, and tumor evasion from the immune system, another death-inducing family member, TNF-related apoptosis-inducing ligand (TRAIL), or Apo-2 ligand, has generated excitement because of its unique ability to induce apoptosis in a wide range of transformed cell lines but not in normal tissues. TRAIL is well tolerated when given to healthy animals, and no observable histological or functional changes have been observed in any of the tissues or organs examined. Moreover, multiple injections of soluble TRAIL into mice beginning the day after tumor implantation can significantly suppress the growth of the tumors, with many animals becoming tumor-free. One potential drawback to these findings is that large amounts of soluble TRAIL may be required to inhibit tumor formation, possibly because of the pharmacokinetic profile of soluble TRAIL that indicates that, after intravenous injection, the majority of the protein is rapidly cleared. Increasing the in vivo half-life of recombinant soluble TRAIL or developing an alternative means of delivery may increase the relative tumoricidal activity of TRAIL such that larger, more established tumors could be eradicated as efficiently as smaller tumors. The information presented here describes the production of an adenoviral vector engineered to carry the complementary DNA (cDNA) for murine TRAIL (hTRAIL). PMID- 19565911 TI - Silencing epidermal growth factor receptor by RNA interference in glioma. AB - Glioblastoma multiforme (GBM) can arise de novo or progress from a lower to higher grade and can possess a series of genetic alterations and dynamic progressions, which have been correlated with the molecular pathology of GBM. Epidermal growth factor receptor (EGFR) has been shown to be overexpressed in a variety of tumors and is one of the important mediators responsible for the development of high-grade gliomas, especially in primary glioblastomas. Most recently, RNA interference (RNAi), in which double-stranded RNA (dsRNA) induces sequence-specific degradation of the targeting messenger RNA (mRNA), has been extensively developed and studied. RNAi is able to silence the targeted gene expression more efficiently and specifically. In the present study, we silence the EGFR expression using two separate short interfering RNAs (siRNAs) targeting the extracellular ligand-binding domain and intracellular tyrosine kinase domain, respectively. We demonstrate that suppression of EGFR expression, by using either antisense or siRNA approaches, inhibits U251 glioblastoma cell growth in vitro and in vivo, and siRNA seems to be more effective than the antisense approach. PMID- 19565912 TI - Delivery of phosphorodiamidate morpholino antisense oligomers in cancer cells. AB - Phosphorodiamidate morpholino oligomers (PMO), which have a neutral chemistry, are extensively being used as tools for selective inhibition of gene expression in cell culture models and are currently in human clinical trials. PMO oligomers possess a unique structure, in which the deoxyribose moiety of DNA is replaced with a six-membered morpholine ring and the charged phosphodiester internucleoside linkages are replaced with neutral phosphorodiamidate linkages. PMO internalization in uptake-permissive cells has been observed to be specific, saturable, and energy-dependent, suggesting a receptor-mediated uptake mechanism. Understanding PMO transport should facilitate the design of more effective synthetic antisense oligomers as therapeutic agents. PMID- 19565913 TI - Use of RNA aptamers for the modulation of cancer cell signaling. AB - Aptamers are in vitro evolved molecules that bind to target proteins with high affinity and specificity by adapting three-dimensional structures upon binding. Because cancer cells exhibit the activation of signaling pathways that are not usually activated in normal cells, RNA aptamers against such a cancer cell specific signal can be useful lead molecules for cancer gene therapy. The Wnt/beta-catenin signaling pathway plays important roles in a critical initiating event in the formation of various human cancers. Because mutations in beta catenin have been found to be responsible for human tumorigenesis, beta-catenin is the molecular target for effective anticancer therapies. Here, we describe the selection of RNA aptamers against beta-catenin/T-Cell Factor (TCF) proteins and their intracellular expression as intramers. The RNA aptamers acted as central inhibitory players for multiple oncogenic functions of beta-catenin in colon cancer cells. These data provide the proof-of-principle for the use of RNA aptamers for an effective anticancer gene therapy. PMID- 19565914 TI - G-rich oligonucleotides for cancer treatment. AB - Oligonucleotides with guanosine-rich (G-rich) sequences often have unusual physical and biological properties, including resistance to nucleases, enhanced cellular uptake, and high affinity for particular proteins. Furthermore, we have found that certain G-rich oligonucleotides (GROs) have antiproliferative activity against a range of cancer cells, while having minimal toxic effects on normal cells. We have investigated the mechanism of this activity and studied the relationship between oligonucleotide structural features and biological activity. Our results indicate that the antiproliferative effects of GROs depend on two properties: the ability to form quadruplex structures stabilized by G-quartets and binding affinity for nucleolin protein. Thus, it appears that the antiproliferative GROs are acting as nucleolin aptamers. Because nucleolin is expressed at high levels on the surface of cancer cells, where it mediates the endocytosis of various ligands, it seems likely that nucleolin-dependent uptake of GROs plays a role in their activity. One of the GROs that we have developed, a 26-nucleotide phosphodiester oligodeoxynucleotide now named AS1411 (formerly AGRO100 or GRO26B-OH), is currently being tested as an anticancer agent in Phase II clinical trials. PMID- 19565915 TI - Regulatory aspects for translating gene therapy research into the clinic. AB - Gene therapy products are highly regulated, therefore moving a promising candidate from the laboratory into the clinic can present unique challenges. Success can only be achieved by proper planning and communication within the clinical development team, as well as consultation with the regulatory scientists who will eventually review the clinical plan. Regulators should not be considered as obstacles but rather as collaborators whose advice can significantly expedite the product development. Sound scientific data is required and reviewed by the regulatory agencies to determine whether the potential benefit to the patient population outweighs the risk. Therefore, compliance with Good Manufacturing Practice (GMP) and Good Laboratory Practice (GLP) principles to ensure quality, safety, purity, and potency of the product, and to establish "proof of concept" for efficacy, and for safety information, respectively, is essential. The design and conduct of the clinical trial must adhere to Good Clinical Practice (GCP) principals. The clinical protocol should contain adequate rationale, supported by nonclinical data, to justify the starting dose and regimen, and adequate safety monitoring based on the patient population and the anticipated toxicities. Proper review and approval of gene therapy clinical studies by numerous committees, and regulatory agencies before and throughout the study allows for ongoing risk assessment of these novel and innovative products. The ethical conduct of clinical trials must be a priority for all clinical investigators and sponsors. As history has shown us, only a few fatal mistakes can dramatically alter the regulation of investigational products for all individuals involved in gene therapy clinical research, and further delay the advancement of gene therapy to licensed medicinal products. PMID- 19565916 TI - Ethics of cancer gene transfer clinical research. AB - Cancer gene transfer is a relatively novel intervention strategy. In part because of this novelty, trials often present greater uncertainties than those investigating more conventional approaches. In the following review, I examine how this greater uncertainty might affect how clinical studies are designed, when they are initiated, their degree of risk, and whether such risk can be justified in terms of therapeutic benefit. The review also discusses two other ethical issues presented by gene transfer clinical research: fairness in subject selection and communications with the public. I conclude with a series of recommendations directed toward researchers, policymakers, and ethics committee members. PMID- 19565917 TI - Virus production for clinical gene therapy. AB - Gene therapy is becoming increasingly relevant for the treatment of prominent human diseases. Viral vectors are currently used in more than 50% of the gene therapy clinical trials, most of them aimed at cancer diseases. Clearly, the increasing needs of high-quality viral preparations require the elimination of process bottlenecks, streamlining the development of a viral vector into a real world clinical tool. Virus production for clinical gene therapy can be a limiting step because many virus generation protocols rely on labor-intensive, bench-scale methods; robust, cost-effective strategies for the delivery of clinical-grade viruses are thus essential for the future of gene therapy. A comprehensive picture of key aspects on the integration of upstream and downstream processing is addressed in this chapter, by describing the case study of recombinant budded baculoviruses for gene therapy; scalable methods are described in detail as well as mandatory characterization techniques for a proper and complete quality assessment of the viral vectors. PMID- 19565918 TI - Production of plasmid DNA as a pharmaceutical. AB - Developments in gene therapy, cell therapy, and DNA vaccination require a pharmaceutical gene vector that, on one hand, fulfils the properties to express the encoded information--preferably at the right place, time, and level and, on the other hand, is safe and productive under good manufacturing practices (GMP). Here we summarize the features of producing and modifying these nonviral gene vectors and ensuring the required quality to treat cells and humans or animals. PMID- 19565919 TI - Gene immunotherapy for non-small cell lung cancer. AB - Historically, limited results have been observed with immunity in non-small cell lung cancer (NSCLC). In the last 5 years, however, several immune-stimulating products have demonstrated enhancement of tumor antigen recognition through activation of dendritic cell-involved processes. Moreover, clinical benefit has been demonstrated in subsets of patients, justifying ongoing phase III investigation. Results of key gene immunotherapies being tested in NSCLC are reviewed. Preliminary results in advanced NSCLC suggest evidence of well tolerated immune activation with suggested evidence of clinical benefit with respect to survival and response. PMID- 19565920 TI - Gene therapy for antitumor vaccination. AB - Tumor immunotherapy depends on the interactions between the host, the tumor, and the immune system. Recent data suggests that priming of antigen-specific T cells alone may not be adequate for mediating regression of established tumors because of the immune inhibitory influences within the tumor microenvironment. Thus, we developed a recombinant vaccinia virus vector to express single or multiple T cell costimulatory molecules as a vector for local gene therapy in patients with malignant melanoma. This approach is feasible and generated local and systemic tumor immunity and induced objective clinical responses in patients with metastatic disease. This chapter reviews the details and major issues related to using live, replicating, recombinant poxviruses for gene delivery and antitumor vaccination within the tumor microenvironment. PMID- 19565921 TI - HSV-TK/IL-2 gene therapy for glioblastoma multiforme. AB - We performed a clinical trial of gene therapy in patients with recurrent glioblastoma multiforme based on retroviral vector-mediated combined delivery of interleukin-2 and thymidine kinase of herpes simplex virus (HSV)-1. Treatment consisted of intratumor injection of retroviral vector-producing cells, followed by intravenous ganciclovir administration. The primary endpoints of the study were the evaluation of the safety of the treatment and demonstration of transduction of tumor cells. Here, we report details on the clinical protocol and the methods used. PMID- 19565922 TI - Construction and characterization of an oncolytic HSV vector containing a fusogenic glycoprotein and prodrug activation for enhanced local tumor control. AB - A large number of oncolytic viral vectors are currently under clinical development for cancer therapy. Herpes simplex virus type 1 (HSV-1) has demonstrated particular promise in this field, showing genetically engineered selective tumor replication and cytotoxicity in a wide variety of tumor types, without damaging healthy tissues. Enhanced activity has been observed when a range of therapeutic genes has been inserted into various oncolytic HSV genomes. Here, we discuss methods used to develop and characterize an oncolytic HSV virus that combines expression of a highly potent prodrug activating gene (yeast cytosine deaminase/uracil phosphoribosyltransferase fusion [Fcy::Fur]) and the fusogenic glycoprotein from gibbon ape leukemia virus (GALV) for enhanced local tumor control. PMID- 19565923 TI - Newcastle disease virus: a promising vector for viral therapy, immune therapy, and gene therapy of cancer. AB - This review deals with the avian paramyxovirus Newcastle disease virus (NDV) and describes properties that explain its oncolytic activity, its tumor-selective replication behavior, and its immune-stimulatory capacity with human cells. The strong interferon response of normal cells upon contact with NDV appears to be the basis for the good tolerability of the virus in cancer patients and for its immune stimulatory properties, whereas the weak interferon response of tumor cells explains the tumor selectivity of replication and oncolysis. Various concepts for the use of this virus for cancer treatment are pointed out and results from clinical studies are summarized. Reverse genetics technology has made it possible recently to clone the genome and to introduce new foreign genes thus generating new recombinant viruses. These can, in the future, be used to transfer new therapeutic genes into tumors and also to immunize against new emerging pathogens. The modular nature of gene transcription, the undetectable rate of recombination, and the lack of a DNA phase in the replication cycle make NDV a suitable candidate for the rational design of a safe and stable vaccine and gene therapy vector. PMID- 19565924 TI - Oncolytic viral therapy using reovirus. AB - Current mainstays in cancer treatment such as chemotherapy, radiation therapy, hormonal manipulation, and even targeted therapies such as trastuzumab (Herceptin) for breast cancer or erlotinib (Tarceva) for non-small cell lung cancer are limited by lack of efficacy, cellular resistance, and toxicity. Dose escalation and combination therapies designed to overcome resistance and increase efficacy are limited by a narrow therapeutic index. Oncolytic viruses are one such group of new biological therapeutics that appears to have a wide spectrum of anticancer activity with minimal human toxicity. Because the malignant phenotype of tumours is the culmination of multiple mutations that occur in several genes eventually leading to aberrant signalling pathways, oncolytic viruses, either natural or engineered, specifically target tumour cells, taking advantage of this cellular deviant signalling for their replication. Reovirus is one such naturally occurring double-stranded RNA (dsRNA) virus that exploits altered Ras signalling pathways in a myriad of cancers. The ability of reovirus to infect and lyse tumours both solid and haematological, under in vitro, in vivo, and ex vivo conditions, is discussed in this chapter. The major mechanism of reovirus oncolysis of cancer cells has been shown to occur through apoptosis. In addition, the synergistic anti-tumour effects of reovirus in combination with radiation or chemotherapy has also been demonstrated for reovirus-resistant and moderately sensitive tumours. In most of the clinical trials undertaken to date, an anti reovirus immune response has been seen likely circumventing efficacy. Investigation into the use of reovirus as an immune adjuvant is currently underway to try and re-direct this immune response to tumour. Reovirus phase I clinical trials have shown indications of efficacy and several phase I/II trials are ongoing at present. The extensive pre-clinical efficacy, replication competency, and low toxicity profile in humans has placed the reovirus as an attractive anti-cancer therapeutic for ongoing clinical testing. PMID- 19565925 TI - Design and testing of novel oncolytic vaccinia strains. AB - Oncolytic or replication-selective viruses have been used as powerful tools for the delivery of therapeutic genes to tumors. Because these vectors are capable of replicating within the tumor, the therapeutic gene is amplified within the target tissue itself, resulting in the spread of the virus both within the tumor, and sometimes also between tumors. Vaccinia virus holds many advantages when serving as the backbone for oncolytic viral strains, including a large cloning capacity (at least 25 kbp) (1); a short life-cycle (2, 3); extensive previous use in humans, with contraindications and adverse reactions well described and antivirals available (4); the potential for systemic (intravenous) delivery to distant tumors; and vaccinia strains have previously demonstrated antitumor benefits in clinical trials (5). Because vaccinia has no known receptor and is capable of infecting almost any cell type, tumor selectivity has to be engineered into vaccinia at steps after infection. We will therefore discuss potential viral virulence genes and metabolic targets that result in tumor-selective vaccinia strains. Because the virus has limited natural requirements for host cell proteins, and, instead, contains a large genome and multiple genes involved in virulence, a large number of possible attenuating gene deletions can result in the production of viral strains reliant on inherent properties of the host cell for replication. The protocols for producing viral gene deletions and constructing viral gene expression vectors have been well established for vaccinia and are summarized briefly in this chapter. Basic assays for testing the tumor selectivity and therapeutic index of new oncolytic constructs in vitro will be covered. In addition, we describe how bioluminescence imaging can be incorporated into preclinical testing of vaccinia gene expression strains to examine the timing, biodistribution, and kinetics of viral gene expression noninvasively after delivery of the viral agents to tumor-bearing mice via different routes. PMID- 19565926 TI - Tumor-targeted Salmonella typhimurium overexpressing cytosine deaminase: a novel, tumor-selective therapy. AB - The ideal anticancer regimen is one that is specific for cancer cells with limited toxicity to normal tissues. Genetically modified, nonpathogenic Salmonella offer a potential way to induce direct tumoricidal activity or to deliver tumoricidal agents to tumors. An attenuated strain of Salmonella typhimurium, called VNP20009, and its derivative TAPET-CD (which expresses Escherichia coli cytosine deaminase) are highly selective for tumor tissue and can deliver therapeutic proteins preferentially to tumors in preclinical models. Both VNP20009 and TAPET-CD have been investigated successfully in Phase 1 clinical trials in cancer patients. PMID- 19565927 TI - Chemoprotection by transfer of resistance genes. AB - Dose-limiting toxicity of chemotherapeutic agents, i.e., myelosuppression, can limit their effectiveness. The transfer and expression of drug-resistance genes might decrease the risks associated with acute hematopoietic toxicity. Protection of hematopoietic stem/progenitor cells by transfer of drug-resistance genes provides the possibility of intensification or escalation of antitumor drug doses and consequently an improved therapeutic index. This chapter reviews drug resistance gene transfer strategies for either myeloprotection or therapeutic gene selection. Selecting candidate drug-resistance gene(s), gene transfer methodology, evaluating the safety and the efficiency of the treatment strategy, relevant in vivo models, and oncoretroviral transduction of human hematopoietic stem/progenitor cells under clinically applicable conditions are described. PMID- 19565928 TI - Phase I clinical trial of locoregional administration of the oncolytic adenovirus ONYX-015 in combination with mitomycin-C, doxorubicin, and cisplatin chemotherapy in patients with advanced sarcomas. AB - Despite many advances in cancer therapy, metastatic disease continues to be incurable in the majority of cancer patients. There is an need for more efficient and less toxic treatments in this setting. Oncolytic virotherapy represents a novel promising direction in the treatment of cancer. Based on preclinical and clinical data, combination with standard chemotherapy has the potential to further increase the antitumor activity of oncolytic virotherapy in a synergistic manner. We present the design of a phase I clinical trial combining intratumoral injections of the oncolytic adenovirus ONYX-015 with systemic chemotherapy in patients with advanced sarcomas. PMID- 19565929 TI - Gastric carcinoma: early detection and prognosis. PMID- 19565930 TI - P53 and vascular endothelial growth factor expressions are two important indices for prognosis in gastric carcinoma. AB - AIM: This study examined the correlation between P53 and vascular endothelial growth factor (VEGF) expression together with tumour vascularity and investigated their clinical significance in the prognosis of gastric carcinoma. SUBJECTS AND METHODS: Ninety-five patients with gastric carcinoma who underwent curative surgical resection were studied using immunohistochemical staining. Correlation between the expression of p53, VEGF microvessel count (MVC) and various clinicopathologic factors were studied. RESULTS: No significant correlation was found between p53 expression and clinicopathologic factors. The rate of VEGF positivity was significantly higher in patients with haematogenous metastasis than in those without haematogenous metastasis. Both p53 and VEGF expression were associated with MVC. The MVC in p53 positive tumours was significantly higher than that in p53 negative tumours. Similarly, the same trend was seen between VEGF expression and MVC. The p53 and VEGF were co-expressed in 61 of 95 tumours (64.2%), and a significant (p < 0.01) association between p53 and VEGF expressions was demonstrated. The rate of VEGF positivity was significantly (p < 0.01) higher in the patients with disease recurrence than in those without recurrence, whereas no significant correlation was found between disease recurrence and the expression of p53. CONCLUSIONS: The p53 expression may play an important role in controlling angiogenesis by regulating VEGF expression and VEGF expression is associated closely with disease recurrence. In addition, both p53 and VEGF expression might be useful in indicating the prognosis in patients with gastric carcinoma. PMID- 19565931 TI - Obstructive lung disease in acute medical patients. AB - OBJECTIVES: To determine the proportion of adult medical patients who have chronic obstructive pulmonary disease (COPD), using the Global initiative for Chronic Obstructive Lung Disease guidelines (GOLD), and its relation to vascular disease. METHODS: This is a prospective cross-sectional study of adult patients admitted to acute medical wards. Interviewer administered questionnaire, anthropometric and spirometric measurements were done. RESULTS: Spirometry was performed in 720 acute admissions [Mean (SD) age 50.0 (18.9) years, FEV1: 1.98 L (0.83), FEV1/FVC%: 75.1 (11.9)%; males 332 (46.1%), smokers 318 (44%); 43.2% had vascular disease]. Sixty-seven per cent of patients (480) had no airway disease including 35 (4.5%) with chronic cough and sputum with normal spirometry; 89 (12.4%) had asthma and 151 (20.9%) had COPD. Patients with COPD were significantly older [60.3 (16.6) years] than non-COPD patients [47.3 (18.5) years], p < 0.001 and had a greater number of pack years of smoking. A greater percentage of patients with COPD had vascular disease (52%) than the non-COPD patients (40.1%), p = 0.017). Multivariate analysis with vascular disease as outcome variable revealed relationships with older age (p < 0.001) and Indo Trinidadian ethnicity (p = 0.015), but not with gender (p = 0.321) and smoking (p = 0.442). FEV1% as well as FEV1 showed a significant inverse relationship with vascular disease (p < 0.05). CONCLUSIONS: The prevalence of COPD using GOLD guidelines in acute hospital admissions is 20.9%; 11.7% of admissions have chronic sputum or cough with normal spirometry. Vascular disease is more prevalent in those with COPD. Patients admitted to acute medical care with vascular disease may also have COPD. PMID- 19565932 TI - Measuring angles on digitalized radiographic images using Microsoft PowerPoint. AB - OBJECTIVE: To introduce the "virtual goniometer", a method of measuring angles on digital images using Microsoft PowerPoint, a readily available and inexpensive software programme. METHODS: Twenty-six X-rays of scoliosis curves were photographed with a digital camera. Six examiners measured the angles of curvature on their computers using the goniometer (Set 1). Under a blinded protocol, repeated measurements on these digitalized X-rays were done three weeks later (Set 2). Intra-observer differences were analyzed. To assess validity, four examiners also measured the angles using the Cobb method. Measurements achieved by both methods were analyzed by the paired samples t-test. To assess inter observer differences, the Pearson correlation coefficient was calculated. RESULTS: Pearson correlation coefficients were significant, r (24) > or = 0.975, p < 0.001. For intraobserver variability, the average 95% CI range was 2.23 degrees between Set 1 and Set 2. The average 95% CI range was 2.38 degrees for the difference between the digital and Cobb methods. CONCLUSIONS: Clinicians using this technique can reliably assume that repeated measurements of scoliosis curvatures will vary in the range of less than 3 degrees. The 95% CI range for intra-observer variability, an index of the technique's repeatability, was > or = 2.4 degrees. A high correlation of measurements can also be expected between different observers with the goniometer. This new technique allows practitioners to utilize an easily accessible computer programme to evaluate angular deformities on digitalized radiographic images accurately and hence reliably make clinical decisions based on these measurements. PMID- 19565933 TI - Pulmonary parenchymal alveolar histological study in experimental tracheo oesophageal malformations. AB - OBJECTIVE: Children operated on for tracheo-oesophageal malformation (TOM) often suffer from postoperative respiratory system difficulties. There is little current literature about this subject. This study aimed to investigate the causes of these problems in rats with experimental TOM by evaluating the lung alveolar histology. SUBJECTS AND METHODS: Twenty Wistar albino rats were used for the experiment. Twelve rats with a sperm positive vaginal smear received 1.75 mg/kg intraperitoneal adriamycin on days six to nine of gestation. A sham group was infused with saline instead of adriamycin. A control group was not subjected to any additional procedure. Their fetuses were dissected under surgical microscope. After examining the trachea and oesophagus, the lungs were dissected and fixed in 10% formalin. The groups were compared with respect to alveolar flat cell (Type 1), capillary density and air space percentage in the samples obtained under light microscopy. Statistical evaluation was performed through Mann-Whitney-U tests and Pearson Chi-squared tests. RESULTS: Type-1 cell ratio and air space percentage were the highest for the control and sham groups. However, the group that received adriamycin and developed TOM had the lowest values. There were no statistically significant differences between the groups with respect to capillary density. CONCLUSION: In rats with experimentally produced TOM, the pulmonary parenchyma showed delayed maturation. This could be the cause of the frequently seen respiratory system pathologies in children suffering from TOM. Further studies should be done to elucidate this. PMID- 19565934 TI - Evaluation of methods and costs for detecting methicillin-resistant Staphylococcus aureus isolates from clinical specimens at regional hospitals in Trinidad and Tobago. AB - OBJECTIVES: To evaluate and determine the most cost effective, rapid and specific method for detection of methicillin resistance in clinical isolates of S. aureus in a setting with limited personnel and resources. METHODS: Standard laboratory methods were used to identify S. aureus isolates. The conventional Methicillin Resistance Staphylococcus aureus (MRSA) detection methods used included, 1 microg oxacillin disk diffusion, oxacillin salt agar screen (CLSI), penicillin binding protein (PBP 2') latex agglutination test and E-tests oxacillin. Results of conventional tests were compared with a polymerase chain reaction (PCR) method for detecting MRSA isolates. Polymerase chain reaction detection of the mecA gene in S. aureus was used as the "gold standard" for MRSA identification. RESULTS: All methods had 100% sensitivity except for oxacillin disk diffusion and oxacillin-salt agar screening with 98% and 99%, respectively. Specificity was also 100% for all methods except for oxacillin-disk diffusion (99%). Turn around time (TAT) for detection of MRSA was calculated to be within six hours for PCR. The fastest TAT of 1.25 hours was obtained for PBP 2' latex agglutination. Total cost for labour and materials to perform each method was highest for E-test, US$13.76/isolate. The cost for PCR when compared to that of latex agglutination was not statistically significant (US$3.74 vs US5.91, p = 0.4). CONCLUSIONS: All methods presented high sensitivity and specificity, but the latex agglutination test had the advantage of giving a reliable, rapid and most cost effective result that compares well to PCR in this environment. PMID- 19565935 TI - Knowledge, perception and practices of healthcare professionals at tertiary level hospitals in Kingston, Jamaica, regarding neonatal pain management. AB - OBJECTIVE: To determine knowledge, perception and practices of healthcare professionals at tertiary level hospitals in Kingston, Jamaica, regarding neonatal pain management. DESIGN AND METHODS: Physicians and nurses actively involved in providing neonatal care at three tertiary level hospitals were invited to participate. A 21-item self-administered questionnaire was used to obtain information on knowledge, perception and practice of neonatal pain management. Descriptive analyses were performed. RESULTS: A total of 147 healthcare workers participated giving a response rate of 85%. Male to female ratio was 1: 4.4. Nurses accounted for 76 (52%) of the respondents while 70 (48%) were physicians. Seventy-three (50%) individuals were unaware of the degree of pain neonates were capable of experiencing and only 38 (27%) knew that premature infants were capable of feeling pain. One hundred and four (71%) respondents were able to identify physiological markers of pain and most respondents were able to discriminate between painful and non-painful procedures. However, 100 (68%) respondents rarely prescribed analgesia for procedures previously rated as painful. Seventy-one (51%) respondents admitted to not using analgesia for alleviating procedural pain in neonates. Twenty-five (18%) individuals thought that the procedure was too short to require analgesic support while 41 (30%) stated that medication was not usually prescribed for procedural pain. Physician scores were significantly higher than those attained by nurses for knowledge (p = 0.003) and for pain perception (p = 0.001) but no significant differences were noted for practice (p = 0.18). CONCLUSION: There is an overwhelming deficiency in the knowledge, perception and practice of neonatal pain management at tertiary level institutions in Kingston, Jamaica. There is the urgent need for the education of health professionals on neonatal pain management. This will in turn facilitate change in perception and eventually, along with the institution of local policies and protocols, influence practice. PMID- 19565936 TI - Association between bone turnover markers and bone mineral density in puberty and constitutional delay of growth and puberty. AB - The aim of this study was to evaluate associations between the markers of bone formation and resorption and bone mineral density in healthy children throughout puberty and in children with constitutional delay of growth and puberty (CDGP). For this reason, 15 boys with CDGP and 75 other boys in different pubertal stages were included in this study. Although mean serum phosphorus level was higher in stages II, III, IV, V compared to stage I and CDGP, mean bone specific akaline phosphatase (b-AP), parathyroid hormone (PTH), bone mineral density (BMD), bone mineral content (BMC) levels were higher in stages III, IV, V compared to stage I and CDGP Mean serum calcium (Ca) levels were lower in stages III, IV, V compared to stage I and CDGP. During puberty, urine DPry/Cr levels were not significant. The peak level of b-AP occurred at stage IV. Serum PTH, Ca, b-AP levels, urine Ca/Cr ratio, BMC and BMD measurements significantly changed during puberty in healthy children. While serum Ca levels progressively decreased, serum b-AP, PTH levels, urine Ca/Cr ratio and bone mineralization increased in healthy children with the level of sexual development. The only significant correlation is found between serum PTH levels and bone mineral density (p < 0.05). In our opinion, PTH may be a potent stimulator of skeletal dynamics in boys and may be associated with substantial increases in lumbar spine. We conclude that PTH behaved as a valuable marker in bone mineralization during puberty. Accelerated bone mineralization is reflected by high levels of serum PTH during puberty. All values of the markers of bone formation and bone resorption in children with CDGP were similar to those of prepubertal children. Children with CDGP had prepubertal properties. We suggest that there is a critical age period for accumulation of bone mass according to the results in this study. PMID- 19565937 TI - Extravasation injuries. AB - OBJECTIVES: To evaluate the time and type of treatment following extravasation from intravenous infusion and the sequelae of the injuries. METHOD: The charts of 12 patients who were referred to the Plastic and Orthopaedic Services at the University Hospital of the West Indies were reviewed. The study period was between May 2003 and January 2007. Data were collected on age, gender, site of extravasation, extravasated agent, treatment of the extravasation, necrosis interval, duration of hospital stay for treatment of injury and whether the intravenous line was resited and at what site in relation to the injury. RESULTS: The age of patients ranged from three days to 67 years. The female-to-male ratio was 2:1. In five patients, the intravenous infusion was discontinued immediately after the swelling was noticed. In two patients, the intravenous infusion was stopped after seven hours and in five patients it was discontinued within 12 to 22 hours. The necrosis interval ranged from 12 hours to three weeks. Immediate treatment following extravasation and discontinuation of the infusion included limb elevation in three patients and application of cold compresses in one patient. Eleven patients developed skin necrosis of varying severities. There was no skin necrosis in one patient. Ten patients spent an average of 31 extra days in hospital for treatment of the extravasation injury. Two patients were treated in an out-patient clinic. CONCLUSIONS: Extravenous leaks can cause severe tissue injuries. Morbidity is increased by delay in recognition and treatment of the extravasation. A protocol for the treatment of extravasation is recommended. PMID- 19565938 TI - Best Practice Guidelines for treatment of hypercholesterolaemia: a statement from the Caribbean Cardiac Society. AB - There is an increase prevalence of chronic non-communicable diseases in the Caribbean as the region undergoes an epidemiologic transition from infectious to chronic non-communicable diseases. Numerous studies have identified hypertension, obesity, diabetes mellitus and hyperlipidaemia as risk factors for the development of coronary atherosclerosis. The Caribbean Cardiac Society recognizes that there is an increased prevalence of these disease entities and in an effort to foster best practice guidelines for the region, implemented a consensus conference for the discussion of hypercholesterolaemia, hypertension, diabetes mellitus and obesity in 2005. This statement outlines the recommendations of the consensus group of the Caribbean Cardiac Society on the Best Practice Guidelines for the therapy of hypercholesterolaemia. PMID- 19565939 TI - The prevalence of mucocutaneous disorders among HIV-positive patients attending an out-patient clinic in Kingston, Jamaica. AB - OBJECTIVES: Skin disorders are thought to occur frequently in persons with HIV/AIDS. To our knowledge, there are no studies in the literature reporting on the spectrum and prevalence of skin disorders in HIV-positive patients in the Caribbean. This study focused on the prevalence and spectrum of skin disorders seen in a population of HIV-positive patients in Jamaica. METHODS: A 5-year retrospective study was conducted by reviewing the records of patients attending a HIV out-patient clinic at the University Hospital of the West Indies in Kingston, Jamaica. RESULTS: Two hundred and eighty-six (286) patients were included in the study. Skin and mucous membrane disorders were documented in 74% of the patients in this series. Inflammatory disorders comprised the largest category of skin disorders followed by fungal infections. The most frequently diagnosed dermatological disorders were papular prurigo, oral candidiasis, dermatophyte infections, herpes simplex infections and seborrhoeic dermatitis. Kaposi's sarcoma was rare. This pattern is similar to those reported from the African continent and other tropical countries. CONCLUSIONS: Dermatological disorders contribute significantly to the morbidity of HIV-positive patients and patterns of skin disorders are similar to those seen in other tropical settings. PMID- 19565940 TI - Suicide in Mie Prefecture, Japan, since 1990. AB - The number of suicides in Japan has increased to over 30,000 per year since 1998. Similarly, the number of suicides has been increasing in Mie Prefecture. In the present study, we examined the incidence and the circumstances of all suicidal cases that were reported to the Mie Prefectural Police Headquarters during the thirteen-year period 1990-2002. In Mie Prefecture, the number of suicides per year averaged 363.1. The largest numbers occurred in the spring and early summer months. For men, suicides were most common in the 50-59-year age group; for women, they were most common in the 70-79-year age group. As for the methods of suicide, hanging was the most frequent for both genders. The major causative factors of suicide were described as "suffering from physical illness", "psychiatric disorders" and "economic difficulties". Of these, "psychiatric disorders" was the most important causative factor for the younger groups of both genders. For the middle-aged group of men, the most important causative factor was "economic difficulties". "Suffering from physical illness" was the most serious causative factor for the elderly group of both genders. In order to prevent suicide, urgent strategies for effective medical treatment and social cooperation are required. PMID- 19565941 TI - Silver nitrate may be far superior to podophyllin in clearing HPV external anogenital warts. AB - Longitudinal data from a case series indicated that clearance rates (95% CIs) of anogenital warts (AGW) by silver nitrate (n = 14) and podophyllin (n = 34) were 93% (79.35, 100) and 14.7% (3, 27) respectively; and RR, 6.31 (95% CI 2.77, 14.37, p = 0.052), relative benefit increase, 532% (410, 654), and NNT, 1 (1, 2). The total effort needed US$28(BB$56) vs US$598 in 1-3 vs 1-37 visits to achieve one successfully treated patient respectively. While subject to the errors of an open, non-randomized case series, silver nitrate was highly efficacious and cost effective and should replace podophyllin at the Winston Scot Polyclinic. PMID- 19565942 TI - Steatohepatitis due to antiretroviral therapy. AB - Jamaica has recorded the largest increase in the rate of HIV/AIDS infection in the English-speaking Caribbean since 1985. Treatment has significantly improved recently with approximately 50% availability of antiretrovirals (ARVs) to patients. The incidence of drug induced hepatotoxicity is not well known for most ARV drugs and few studies have assessed adverse drug effects in clinical practice. A patient with HIV on highly active antiretroviral therapy (HAART) presented with a one year history of progressive abdominal distension. Abdominal examination revealed a 17 cm, smooth, non-tender liver with a rounded edge; 12 cm of which was below the right costal margin. Liver enzymes were grossly abnormal. The liver biopsy revealed parenchymal distortion by fibrosis with macrovesicular fatty change and Mallory's hyaline in keeping with steatohepatitis. Follow-up studies after discontinuation of stavudine revealed that the liver enzymes improved within four months. Physicians should be mindful of the hepatotoxic potential of ARVs and monitor liver enzymes in HIV-infected patients on therapy. PMID- 19565943 TI - Acinic cell carcinoma of the parotid gland in children: a case report and literature review. AB - Parotid acinic cell carcinoma is a rare malignancy in childhood. We report the case of a 12-year-old girl presenting with a palpable mass in the left maxillofacial area. The radiologic evaluation showed a parotid mass. Tumour resection revealed acinic cell carcinoma of the parotid gland. She underwent complementary total parotidectomy without any adjuvant treatment. The patient has been disease-free for the last five years. We review the literature on acinic cell carcinomas of parotid glands in childhood. PMID- 19565944 TI - Famine and war. AB - History records that mankind has suffered at intervals from great famines. As recently as 1943, at least a million people died of famine in Bengal. In January 1943, a lot of soldiers of the Royal Hungarian Army died in Russia, at the River Don, because of the lack of food supply and insufficient healthcare. The enormous advances of science in this century have made the prevention and relief of famine technically easier. Subsequently, in many parts of the world, famine conditions have arisen, but fortunately on a smaller scale. In this work an attempt is made to set out a few general principles of famine. PMID- 19565945 TI - Herbal preparation use among patients with benign prostatic hyperplasia attending a urology clinic in Jamaica, West Indies. PMID- 19565946 TI - Suicide: a preventable public health problem. PMID- 19565947 TI - Age-specific incidence of cancer in Kingston and St Andrew, Jamaica, 1998-2002. AB - A total of 4737 cancers were recorded in Kingston and St Andrew during the period 1998-2002: 2387 in males and 2350 in females. Age standardized rates per 100,000 per year (ASR) were 188.6 and 144.2 for males and females respectively. In males, the leading sites for cancer were prostate (873 cases), bronchus (269 cases) and large bowel (176 cases) while in females, the leading sites were breast (639 cases), cervix uteri (318 cases) and large bowel (218 cases). The number of new cases has remained relatively stable, compared to the previous five-year period (1993-1997), but the ASR for females has fallen from 156.7 to 144.2 per 100,000 per year and that for males has increased from 176.7 to 188.6 per 100,000 per year. The leading sites of cancer for both males and females have also been maintained in the same order but there was a moderate increase in prostate cancer (ASR 56.4 vs 65.5 per 100,000 per year) and a moderate decrease in cervical cancer (ASR 25.2 vs 19 per 100,000 per year). The incidence of cancer of the female breast has remained relatively stable (ASR 43.2 vs 40.1 per 100,000 per year). In both males and females, colon cancer incidence rates remain high at ASR of 9.9 per 100,000 per year in males and 9.4 per 100,000 per year in females. These data support the continuation of existing programmes for prevention and control of cancers of the prostate, lung, breast and cervix uteri and the introduction of new programmes specifically for cancers of the large bowel in the Jamaican population. PMID- 19565948 TI - Clinicopathologic features of breast disease in Jamaica: findings of the Jamaican Breast Disease Study, 2000-2002. AB - OBJECTIVE: To describe the clinicopathologic profile of breast disease in Jamaica. METHODS: The Jamaican Breast Disease Study is an ongoing prospective, multidisciplinary investigation of breast disease at the University Hospital of the West Indies (UHWI). The initial phase was a prevalence survey comprising all consenting patients referred to the Surgical Outpatient Department (SOPD) UHWI, for breast disease. Demographic, clinical, radiologic and pathologic information were recorded for each patient and the data for the first three years (2000-2002) were analyzed. RESULTS: A total of 1189 patients was enrolled for the study period (28.8% of all new SOPD patients). The age range was 10 to 93 years (mean/SD = 36.5 +/- 16.4 years) with a female : male ratio of 14:1. Most patients (67.8%) presented with a palpable lump and the clinical diagnosis was benign in the majority (70.4%) of patients. Fibroadenoma was the most common benign histologic result (39.4% of all biopsies) followed by non-proliferative (fibrocystic) disease (19.3% of all biopsies). Proliferative disease without atypia, complex fibroadenoma and atypical ductal hyperplasia accounted for 6.9%, 2.6% and 0.4% of biopsies respectively. Overall, 23.4% of biopsies showed malignant histology (10.8% patients); invasive ductal carcinoma accounted for the majority of these cases (69.5%). CONCLUSIONS: The majority of patients with breast disease in Jamaica are young women with clinically benign disease. There was a low prevalence of clinically significant premalignant disease. This is the first study to prospectively describe the clinicopathologic features of breast disease in Jamaica and supports the need for advocating breast cancer screening to facilitate detection of significant premalignant disease and early stages of breast cancer. PMID- 19565949 TI - Trends in the prevalence of adolescent births in Antigua and Barbuda over 35 years. AB - A retrospective study was undertaken to determine trends in teenage births in Antigua and Barbuda from 1969 to 2003. Maternity Ward records were reviewed at Holberton Hospital, site of over 90% of deliveries in Antigua and Barbuda. Maternal age for all births were included. The estimated population in Antigua and Barbuda increased from 60,159 in 1969 to 69,866 in 1998, with no suspected change in age-group or gender distribution. Births to mothers 12-13, 14-15, 16-17 and 18-19 years old were analyzed separately. Yearly birth totals for the 10-year period from 1969 to 1973 and 1999 to 2003 were compared using student t test analysis. The total number of births averaged 1075 per year with no change in trend. For females aged 12 and 13 years, there was no change in births from 1969 to 1978 compared with 1994 to 2003. For females aged 14 years, there was a 54% drop; for females aged 15 years, a 59% drop; for females aged 16 years, a 53% drop; for females aged 17 years, a 51% drop; for females 18 years, a 37% drop and for females aged 19 years, a 30% drop, while births to mothers > or = 20 years increased 25%. The decrease for all teen births was 42% from 1969-1978 to 1994 2003. Teen births accounted for 29% of all births at Holberton in 1969 to 1973 but only 16% of births in 1999 to 2003. In conclusion, Antigua and Barbuda has seen a marked drop of 42% in teenage deliveries, including a 53% drop in school aged teens from 1969 to 2003. Explanations include factors such as economic, social, educational and medical developments. PMID- 19565950 TI - Urinary tract infection in neonates with serious bacterial infections admitted to the University Hospital of the West Indies. AB - OBJECTIVE: The aim of this study was to describe the epidemiology of urinary tract infection in neonates, with serious bacterial infections, admitted to the University Hospital of the West Indies. METHODS: Admission records of all neonates admitted to the neonatal unit of the University Hospital of the West Indies between January 1995 and December 2000 for sepsis evaluation were reviewed. Infants who had positive bacterial cultures were identified using laboratory records. Those satisfying the criteria for urinary tract infection were selected for detailed analysis. Demographic, clinical and laboratory data were collected using a pre-coded questionnaire. Descriptive analyses were performed. RESULTS: Fifty-one (38%) of 135 babies with positive bacterial isolates had confirmed urinary tract infection. Male:female ratio was 6:1. Common presenting features included fever (32%), poor feeding (30%) and irritability (22%). The mean white cell count was 14 +/- 6.26. E coli and Klebsiella species were most frequently identified. Factors associated with a diagnosis of urinary tract infection included male gender (p < 0.001), age > 48 hours (p < 0.05) and a presenting complaint of poor feeding (p < 0.003). Imaging studies of the renal tract detected abnormalities in 5 (10.4%) neonates. CONCLUSIONS: Urinary tract infection is an important cause of serious bacterial infection in neonates affecting 1 in 3 babies with proven bacterial infection and may be the first indicator of underlying structural abnormalities. The absence of specific distinguishing clinical features makes it necessary to include urine cultures in the sepsis evaluation of neonates presenting with symptoms suggestive of sepsis. PMID- 19565951 TI - Activity of amikacin, ertapenem, ciprofloxacin and levofloxacin alone and in combination against resistant nosocomial pathogens by time-kill. AB - OBJECTIVE: The purpose of this study was to determine the synergistic activity of amikacin/ertapenem, fluoroquinolones (ciprofloxacin and levofloxacin)/ertapenem and amikacin/fluoroquinolones combinations against resistant nosocomial pathogens. METHODS: Time-kill studies were performed over 24 hours using an inoculum of 5 x 10(6) - 1 x 10(7) cfu/mL. Antibiotics were tested at the 1 x MIC and 4 x MIC concentrations. RESULTS: At MIC and/or 4 x MIC concentrations, the antibiotic combinations showed additive or synergistic activity against Acinetobacter strains and extended spectrum beta-lactamase producing Klebsiella pneumoniae. In Escherichia coli strains, synergy was seen when amikacin was combined with ertapenem, ciprofloxacin and levofloxacin; ertapenem in combination with fluoroquinolones demonstrated antagonism. In Pseudomonas aeruginosa strains, synergistic effect was exhibited by ertapenem plus amikacin and ertapenem plus fluoroquinolones. The antibiotic combinations showed antagonistic interaction in methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecalis. CONCLUSION: The antibiotic combinations showed additive or synergistic activity against many gram-negative pathogens. PMID- 19565952 TI - Utilization pattern and cost of sedation, analgesia and neuromuscular blockade in a multidisciplinary intensive care unit. AB - OBJECTIVES: To study the utilization pattern and the cost of sedatives, analgesics and neuromuscular blocking agents in a multidisciplinary intensive care unit (ICU). METHODS: A prospective observational study was conducted in the ICU of the Eric Williams Medical Sciences Complex, Trinidad and Tobago, for a period of twelve weeks. All patients admitted to the ICU were enrolled. No interventions were done. Data collected included demographics, diagnoses on admission, length of stay in the ICU, status of mechanical ventilation, patient outcome, quantity of sedatives, analgesics and neuromuscular blocking agents used in every patient and their cost. RESULTS: There were 333 patient-days encountered from 34 patients studied. Midazolam, fentanyl and cisatracurium were the most commonly used sedative, opioid and neuromuscular blocking agents respectively. The total cost of drugs used for sedation, analgesia and neuromuscular blockade was approximately US$ 19,600 per annum. Cost for this treatment alone accounted for more than 50% of the total ICU drug costs. The costs were significantly higher in patients who stayed more than two weeks in the ICU when compared to those who stayed less than two weeks (p < 0.001). CONCLUSIONS: The study highlights the utilization pattern and financial burden of sedation, analgesia and neuromuscular blockade in the delivery of critical care. PMID- 19565953 TI - Systemic sclerosis in an Afro-Caribbean population: a review of demographic and clinical features. AB - OBJECTIVE: To assess the clinical and selected demographic features of patients with systemic sclerosis (SS) seen over a 10-year period at the Rheumatology service of the Queen Elizabeth Hospital, Barbados. To compare these data with what is known to obtain in other ethnic populations. DESIGN AND METHODS: A chart review involving all patients who were found to have SS based on the American College of Rheumatology clinical criteria was conducted between 1996 and 2006. RESULTS: Twenty-seven patients with SS were identified in this predominantly Afro Caribbean population. The prevalent and incident cases numbered 10 and 17 respectively. Twenty-six of these patients were female and the mean age at diagnosis was 37.3 years. Diffuse cutaneous involvement was seen in 63% of cases and limited cutaneous involvement in 37%. The most common clinical features in descending order of frequency were Raynaud's phenomenon, gastroesophageal reflux, pigmentary skin changes, digital pitting/ulceration, telangiectasia and pulmonary disease. CONCLUSION: In a predominantly Afro-Caribbean population, SS was uncommonly seen, had a marked female preponderance and an earlier age of onset than that seen in Caucasian populations. As expected, diffuse disease was the more common subtype and digital pitting, pigmentary skin changes, and pulmonary disease were amongst the most frequent clinical features. Telangiectasia were found more frequently than the literature suggests is typical for patients of African descent. PMID- 19565954 TI - Mortality trends and potential years of life lost in the English and Dutch speaking Caribbean, 1985-2000. AB - OBJECTIVE: To describe mortality trends and potential years of life lost (PYLL) due to leading causes of death in 21 Caribbean countries during 1985, 1990, 1995 and 2000. METHODS: Mortality data for 1985, 1990, 1995 and 2000 were analyzed to identify regional mortality trends using crude, age-specific and age-adjusted death rates and potential years of life lost. The variables used were age, gender and underlying cause of death. RESULTS: During 1985-2000, there was an overall 5% decrease in age-adjusted mortality rates and male mortality exceeded female mortality. Heart disease was the leading cause of death, with cancers, cerebrovascular diseases, diabetes mellitus and hypertensive disease also among the top five causes in most years. Prostate cancer and cancer of the uterus and breast were the leading causes of death due to cancers. HIV disease (AIDS) featured in the ten leading causes of death for the first time in 1995 and was the 5th leading cause in 2000. CONCLUSION: During the period 1985-2000, countries experienced an increase in mortality due to non-communicable diseases, AIDS and assaults (homicides); the latter two causes were most common among the 15-44 year age group. In 2000, AIDS, heart disease and assault (homicide) were the largest contributors to PYLL. PMID- 19565955 TI - Acetaminophen overdose in Jamaica. AB - Acetaminophen is a dose dependent hepatotoxin which is frequently associated with intentional self-harm. Forty-nine cases of parasuicide attempts involving paracetamol only or in combination with another drug were treated at the UHWI, Jamaica, between 1994-2004. The majority were women (84%) and the mean age was 23 years. Acetaminophen was the only agent ingested in 71% of cases; 29% involved an additional drug. Patients presented an average of 6.5 hours after ingestion (range 1-45 hours). Serum transaminases were elevated in 18% of cases and N acetylcysteine (NAC) therapy given in 55%. The mean duration of hospitalization was three days. One patient developed liver failure and there were no deaths. Education of the public and medical profession is needed to increase awareness of the potential toxic effects of acetaminophen overdose. N-acetylcysteine therapy should be given early in suspected cases. PMID- 19565956 TI - Evaluation of the validity of the Quality of Well-being Scale in Trinidad and Tobago. AB - Both developing countries in the Caribbean and developed countries face resource allocation challenges. However, cost-effectiveness analysis instruments that may assist in allocation of resources have not been tested in Caribbean countries. Trinidad and Tobago is an advantageous location to test an instrument for potential use in the Caribbean. It has a single payer healthcare system and a literate population. Due to historical and current migration from other Caribbean countries, the population might be a fair representation of English-speaking Caribbean nations. We tested the validity of the Quality of Well-being Scale (QWB) on a sample of the non-institutionalized general population in Trinidad. The survey included reports of chronic conditions and items from the Trinidad and Tobago National Health Interview Survey. Data were analyzed using a multivariable regression model. One adult from each of 235 households consented to the interview. The results are consistent with results obtained in the United States of America. Being older female, more chronic conditions and more symptoms/problems were significantly associated with lower mean QWB scores. These results suggest that the QWB with US-derived weights show evidence of validity in Trinidad and Tobago. Thus, health decision makers can use the QWB to compare the effects of different health conditions and health interventions. In addition, investigators can make cross-cultural comparisons of QWB scores for diseases or health conditions. PMID- 19565957 TI - The impact of gender, family and type of school on smoking in adolescents in Eyup, Istanbul, Turkey. AB - OBJECTIVE: The aim of this study was to determine the influence of smoking habits and education of family, parents, the type of school and gender on smoking in adolescents. METHODS: This cross-sectional study was performed on 1062 students from eight high schools in Eyup, Istanbul. RESULTS: Twenty-five per cent of students were current smokers (30.1% of boys [n = 551], 20.9% girls [n= 511] 95% CI [26%, 34%] and [17%, 24%] respectively, (p < 0.05). Smoking rate was highest in vocational high schools and lowest in super high schools (33% [n = 406, 95% CI (29%, 38%)] and 11% [n = 127, 95% CI (6%, 17%)] respectively, p < 0.05). Smoking rate was higher in mothers (31.6% [n = 174, 95% CI (25%, 39%)] vs 15.5% [n = 336, 95% CI(12%, 19%)] and siblings (27.7% [n = 141, 95% CI (20%, 35%)] vs 18% [n = 350, 95% CI (14%, 22%)] of the female students who smoke than of those who did not smoke (p < 0.05). The rate of smoking in the siblings of 'smoker' boys was higher than that of 'non-smoker' boys (44.6% [n = 130, 95% CI (36%, 53%)] and 26.1% [n = 402, 95% CI (22%, 30%)] respectively; p < 0.05). The education level of 28%, [n = 82, 95% CI (19%, 39%)] of the mothers of 'smoker' girls, and of 19.6% [n = 429, 95% CI (16%, 23%)] of 'non-smoker' girls was high school and above (p < 0.05). The rate of high school and above education was higher in parents of male students who smoke than that in nonsmokers [45.1% and 27.9%, 95% CI (33%, 55%)] and (24%, 32%), respectively, p < 0.05. Number of smokers saying "Yes" in support of controlling smoking was less than non-smokers (p < 0.05). CONCLUSION: Smoking rate was higher in boys, in vocational high schools and seems to be influenced by smoking habits and higher educational levels of their family members. This influence differs according to gender. Anti-tobacco messages should target family, friends and schools of adolescents. PMID- 19565958 TI - White blood cell counts in healthy Jamaican adults. AB - The investigation of presumed neutropenia places a burden on the health services, especially those of developing countries, including Jamaica. This may be because the normal ranges used in the laboratory are based on the values generated from the Caucasian population. Previous studies looking at African and Afro-Caribbean groups have found lower counts for these populations compared with Caucasians. To address this issue, 195 healthy adults donating blood at the National Public Health Laboratory and the University Hospital of the West Indies blood banks in Kingston, Jamaica, were screened for complete blood count (CBC) differentials between June 2001 and June 2006. The geometric means for the neutrophil counts were found to be 2.4 x 10(9)/L for men and 2.7 x 10(9)/L for women, with 95% confidence intervals of 2.2-2.8 x 10(9)/L and 2.5-3.1 x 10(9)/L respectively. Values for the Jamaican population were similar to those of other Afro-Caribbean groups. Based on this distribution, 14% of healthy Jamaicans would fall below the normal ranges derived from Caucasians and therefore presumed to have neutropenia. We recommend that the lower reference ranges obtained for Afro-Caribbean adults be adopted for that population. PMID- 19565959 TI - Market survey of loiasis: prevalence and adverse reactions to ivermectin using a rapid procedure for loiasis. AB - OBJECTIVES: 1. To assess the levels of endemicity of loiasis in a total of 30 selected villages covering an area of 5.2 sq km in Owo, Akure North, Akure South, Ondo East, Ondo West and Ifedore local governments in Ondo State, Nigeria, using Rapid Assessment for loiasis technique. 2. To obtain information on adverse reactions which occurred during previous administrations of the drug ivermectin used for the treatment of loiasis and onchocerciasis in the study area. METHODS: Consent was obtained from the chairman of each local government village head and individuals were co-opted into the study. A total of 480 individuals were interviewed, 80 from each of the six local governments, during the regular scheduled community market days. Survey for Loa loa was carried out according to specifications in "Guidelines for rapid Assessment of Loa loa (TDR, 2002). RESULTS: Among the 480 respondents, 80 (16.7%) reported having had a history of loiasis while 11 (2.3%) had recent occurrence of the disease. The local name given to the disease in all the villages studied is "aran oju". Respondents in Ondo East local government had an eyeworm history of 23 (28.8%) and occurrence of loiasis was 02 (2.5%), these were the highest figures recorded. Using the SPSS version 10 package for data analysis, demographic factors of age and gender were not significant determinants of prevalence (> 0.05). Ondo East and Ifedore local governments had significant levels of loiasis prevalence (p < 0.05). Adverse reactions from the retrospective ivermectin administration was 04 (0.83%). This was statistically insignificant (p > 0.05). CONCLUSION: Market surveys in rural communities were considered to be cost effective and rapid in the determination of prevalence of any disease. Since less than 40% of the respondents in the study area reported having had a history of eyeworm, the entire area was classified as "low risk" as regards the administration of ivermectin. PMID- 19565960 TI - Diagnosis of Behcet's disease in a non-endemic region: the utility of the urate skin test. AB - Sporadic cases of Behcet's disease in non-endemic regions pose a diagnostic challenge and may be confused with other more common chronic, relapsing multisystem disorders. The urate skin test, an exaggerated inflammatory reaction to intradermally injected monosodium urate crystals, may add a level of diagnostic specificity to a disease which otherwise lacks pathognomonic clinical features. PMID- 19565961 TI - The evolution of emergency medicine in Jamaica. AB - Emergency Medicine in Jamaica has evolved rapidly over the past 20 years and has gained recognition as a specialty. A residency training programme has been established, trademark life support courses are now available, moves to develop areas of sub-specialization have begun and an emergency medicine association has been formed. There has been an increase in the diagnostic modalities in the main teaching institution, the University Hospital of the West Indies (UHWI). There is an urgent need for improvements in pre-hospital care. This will require the development of an efficient Emergency Medical Service (EMS). More emphasis and attention is required on disaster medicine, toxicology and trauma. Increased training of emergency physicians and nurses, advances in academia and research, and greater advocacy by local emergency physicians will further advance the specialty. PMID- 19565962 TI - Single photon emission computerized tomography and histological evaluation in the validation of a new technique for closure of oro-antral communication: an experimental study in pigs. AB - Various bone regeneration techniques have evolved recently but controversies regarding vascularization and integration of such bone grafting techniques have led occasionally to animal experiment to validate such techniques. The objective of this study was to evaluate the evidence of vascularization and osseo integration of a new bone regeneration technique utilized for the closure of oro antral communication (OAC) by an experimental model in which Single Photon Emission computerized Tomography and histological studies were conducted in pigs. We conclude that the sandwich technique used for the closure of OAC results in a vascularized new bone formation which eventually osseo-integrate with the surrounding bone. Also, this experimental study confirmed that autogeneous bone graft was superior to xenografts when used within the sandwich unit. PMID- 19565963 TI - Subclinical hyperthyroidism presenting with bradycardia-associated syncope. PMID- 19565964 TI - Delayed development of a biloma in an infant following hepatic injury. AB - A case report is presented of an infant who developed a biloma over three months after major hepatic injury and after almost complete healing. A brief literature review is given to highlight unusual features of this case. PMID- 19565965 TI - A case of pathological rib fractures: focal osteolysis or osteoporosis? AB - This paper reports on a unique, previously unreported, successful outcome in the case of a patient with focal osteolytic lesions of the ribs as a first sign of osteoporosis. The lesions were detected by chance after acute cough-induced rib fractures were seen on plain chest radiographs. The diagnosis had to be approached as a diagnosis of exclusion since known causes of the osteolytic process had to be eliminated. The authors describe multiple focal osteolytic lesions with rib fractures appearing in a pattern that could be confused with metastases. Laboratory results were normal. Final diagnosis was based on plain radiography, bone scan and bone densitometry. Pharmacomedical treatments for osteoporosis were applied. The patient was observed between the year 2000 and 2005. Five years later radiological and bone scintigraphy revealed resolution of the lesion. We conclude that osteoporosis should be included in the differential diagnosis of asymptomatic focal osteolysis of the ribs with rib fractures as a complication of acute cough. The case suggests that focal osteolytic lesions of the ribs may regress over time and become scintigraphically inactive. PMID- 19565966 TI - Changes in the divorce and birth rates correlate with the suicide rate in Japan. PMID- 19565967 TI - The correlation between unemployment and suicide rates in Gifu Prefecture, Japan. PMID- 19565968 TI - Present and future subjects of suicide-related reports in Japan. PMID- 19565969 TI - The need to strengthen malaria control in the Caribbean in the era of HIV/AIDS. PMID- 19565970 TI - Chemical composition of urinary tract stones at the University Hospital of the West Indies. AB - Urinary tract stones occur frequently with the incidence being about one to fifteen per cent worldwide. Patients may be asymptomatic or sometimes they may present with haematuria. Severe lumbar pain radiating to the loin requiring immediate analgesic treatment may occur Stones generally consist of organic and inorganic material. The organic material may be present in the nidus and can contribute up to about 2.5% of the total weight. Inorganic minerals make up the bulk of the stone. Data are presented for the inorganic minerals present in the stones seen at the University Hospital of the West Indies over a 25-year period. Six hundred and forty-one (445 males and 196females) stones were analyzed by routine chemical methods. Calcium was the main constituent, being seen in 93.9% of the stones. This was followed by oxalate 60.1%, urate 37.0%, bicarbonate 16.5% and magnesium 8.6%. There were four cystine containing stones. Treatment includes medical management for the underlying cause and surgical methods for the removal of the stones. Chemical methods of analysis of the stones has its limitations and should be replaced by more sophisticated methods eg X-ray diffraction crystallography which would give more accurate details of the structure of the stones. PMID- 19565971 TI - An analysis of dietary micronutrient intakes in two age groups of black South African women. AB - OBJECTIVE: To assess micronutrient intake of black women living in Mangaung, South Africa. SUBJECTS AND METHOD: A sample of 500 pre-menopausal black South African women (496 qualified to participate) from two age groups (25-34 and 35-44 years) were selected randomly in Mangaung, the black residential area of Bloemfontein. A validated Quantitative Food Frequency Questionnaire (QFFQ) was used to determine dietary intake of participants. Data were categorized into the two age groups. Median micronutrient intakes were compared to the Recommended Dietary Allowance (RDA) and Adequate Intake (AI). The prevalence of women with intakes < or = 67% of the RDA was calculated. RESULTS: Median calcium and vitamin D intakes were lower than the AI. Of all women, 46.2% to 62.2% consumed < or = 67% of the RDA for total iron, selenium, folate and vitamin C, and more than 94% consumed < or = 67% of the RDA for selenium. At least 25% of all women consumed < or = 67% of the RDA for vitamin A and E. The vitamin B6 intake of older women was inadequate and a fairly large percentage of the total sample consumed < or = 67% of the RDA. CONCLUSION: Generally, micronutrient intakes were adequate in this population. Attention should be given to those micronutrients where median intakes were < or = 67% of the RDA and those that were not at or above the respective AI in these groups of women. PMID- 19565972 TI - The nutritional status of clinic attendees living with HIV/AIDS in St Vincent and the Grenadines. AB - OBJECTIVES: The purpose of this study was to assess nutritional status and dietary practices in persons living with HIV/AIDS (PLWHA). METHODS: A case control design was used. Cases consisted of 36 PLWHA. Controls consisted of 37 persons within the same age range from the general population. Participants filled out a questionnaire consisting of sociodemographic, dietary and health history items. In addition, they had weight, height, upper mid-arm circumference and triceps skinfold measured using standard procedure. Biochemical and clinical data for cases were extracted from their clinic file. RESULTS: HIV-positive persons had significantly lower mean weight, BMI, upper mid-arm circumferences, arm muscle area and arm fat area than persons in the control group. They were also less likely to use multivitamins, dietary supplements, fruit and vegetables than persons in the control group. Correlation coefficients between corrected arm muscle area (CAMA) and BMI and weight ranged from 0.67 to 0.74 in cases and 0.41 to 0.68 for the control group, respectively. Screening for depleted CD4 counts using gender specific CAMA cut-offs indicative of depleted arm muscle reserves resulted in 48% sensitivity and 100% specificity in identifying PLWHA with CD4 counts < 200 cells/microL. CONCLUSION: The findings suggest that PLWHA are at increased risk for poor intakes of fruits and vegetables and depleted lean body mass. In addition, CAMA along with other clinic measures might be useful in the identification of PLWHA who might be responding adequately to treatment. PMID- 19565973 TI - HIV and syphilis infection among gold and diamond miners--Guyana, 2004. AB - BACKGROUND: Guyana had an estimated HIVprevalence of 1.5% among pregnant women in 2006 (95% confidence interval [CI] = 1.1-1.9). However, a survey of miners in one mine found a 6.5% HIV prevalence in 2002. To determine whether Guyanese miners are at high risk for HIV infection we conducted a HIV and syphilis prevalence survey of miners in several mines. METHODS: Adult male consenting miners in 45 Guyanese mines were interviewed, counselled, tested for HIV and syphilis with rapid tests and provided onsite test results. The survey was cross-sectional and used a multi-stage cluster sampling design; population estimates were calculated using SUDAAN. RESULTS: Of 651 miners approached, 539 (83%) were interviewed and 509 (78%) tested. The estimated prevalence for HIV was 3.9% (CI = 2.1, 7.1) and for life-time syphilis exposure was 6.4% (CI = 4.5, 9.1). Fifty-four per cent (CI = 41.3, 66.7) of miners had casual sex during the preceding year, of whom 44.4% (CI = 34.3, 55.0) had always used condoms with these partners. CONCLUSION: The estimated HIV prevalence among Guyanese miners was higher than that of the general population. Targeted interventions including condom promotion are recommended to prevent further spread of HIV and other sexually transmitted infections among miners. PMID- 19565974 TI - Determinants of risk behaviour of sex-workers in Jamaica. A qualitative approach. AB - The practice of evidence-based medicine is well established and research is a key element. Evidence-based approaches require an understanding of the complexities of the interactions and dynamics that influence decision-making. Qualitative research methods are increasingly being used to complement the findings of the traditional quantitative methods. A qualitative study which examines the sexual choices of commercial sex-workers is described to demonstrate the usefulness of this type of study. Focus group discussions give the reasons for the non-regular use of condoms and identify other factors needing intervention. These include low self-esteem and poor job satisfaction as well as problems with violence and drug use. It provides vital information for HIV/AIDS prevention activities for commercial sex-workers. PMID- 19565975 TI - Malaria control and treatment strategies among school children in semi-urban tropical communities. AB - BACKGROUND: Plasmodium infections among school children in Igbo-Eze South Local Government of Enugu State, Nigeria, were studied between July and December 2005. The relationship between the use of malaria control measures and the prevalence of Plasmodium infections was investigated. METHODS: Blood smears were obtained from 1296 school children (ages 4-15 years) from six schools randomly sampled from the study area. Drugs were given to infected children. Epidemiological questionnaires were administered to the children. RESULTS: Out of 1296 school children examined, 270 (20.8%) had Plasmodium falciparum infections. The prevalence of these parasitic infections varied significantly (p < 0.05) among schools, with Central School, Ovoko (30.1% P falciparum) and Community primary school, Itchi (13.9%), having the highest and lowest prevalence rates respectively. Furthermore, the prevalence of P falciparum infections also varied significantly (p < 0.05) among the age groups, with age groups 4-6 (35.1%) and 10 12 (14.2%) having the highest and lowest prevalence rates respectively. Males (23.1%) had a significantly higher prevalence rate than females (18.5%). The prevalence of malaria was significantly lower among pupils using preventive measures; 5.9% among pupils using mosquito bed net as against 21.2% among those not using bed nets and 4.6% for pupils living in screened houses as against 24.1% for those not living in screened houses. CONCLUSION: The study revealed that malaria is a major public health problem among pupils in a Nigerian local community. Prevalence rates among pupils varied among location of schools, age and gender. Preventive measures favoured the use of bed nets and living in screened houses. PMID- 19565976 TI - Malaria and its vectors in the Caribbean: the continuing challenge of the disease forty-five years after eradication from the Islands. AB - OBJECTIVES: Given the occurrence of autochthonous malaria in non-endemic island countries in the last 10 years, this study evaluates the risk factors for malaria transmission in the malaria "endemic and "non-endemic" countries of the Caribbean region. DESIGN: Data on imported and autochthonous malaria for the 27-year period (1980-2006) were gathered from surveillance units in the 21 Caribbean Epidemiology Centre (CAREC) Member Countries (CMCs) via the CAREC epidemiology unit. Anopheles mosquito data were also gathered from various sources. The vector and malaria data were correlated to determine the current risk of malaria transmission. RESULTS: Imported cases. For the 26-year period (1980-2005), there were 897 reported cases in the CMC islands. Jamaica (38.4%) > Trinidad and Tobago (19.5%) > Bahamas (15.8%) > Cayman Islands (12.5%) were mostly affected. Only the smallest CMCs eg Anguilla and British Virgin Islands reported no imported malaria. Indigenous malaria. Over the same time period, malaria was seen mainly in the three mainland countries of Guyana (514 386 cases) > Suriname (275 361) > Belize (85 313). However, for the period 1995-2005, Belize and Guyana reported reduction in case numbers of 84% and 54% respectively. At the same time, Suriname reported a cyclical pattern of reported cases resulting in 77% increase in cases between 1995 and 2005. "Non-endemic" CMCs such as Trinidad and Tobago, and Bahamas, did report autochthonous malaria. In 2006/7, Jamaica reported 340 P falciparum cases, coming just 1-2 years after a massive 505% increase in imported malaria in the region--88% in Jamaica. Anopheles spp: There was a rich diversity of Anopheles mosquitoes--29 spp. in CMCs. Mainland CMCs and nearby island countries had most spp. recorded. Smaller countries with limited ecological niches such as St Kitts, Anguilla, Turks and Caicos Islands (TCI) and Bermuda had little or no Anopheles spp. Two main Anopheles axes were identified--An albimanus in the northern CMCs and An aquasalis in the southern Caribbean. CONCLUSION: All the essential malaria transmission conditions--vector, imported malaria organism and susceptible human host--now exist in most CMCs. A call is now made for enhanced surveillance, vector control and anti-malaria skills to be established in CMCs, in particular in: Recognizing the possible impact of climate change on the spread of anopheles and malaria transmission. Improving vector control skills for anopheles in CMCs. Strengthening malaria surveillance skills. Upgrading malaria therapy and prophylaxis. Emphasizing malaria prevention and educationfor all community and professional sectors. PMID- 19565977 TI - Impact of environment and behaviour on self-esteem in Jamaican adolescent girls. AB - The goals of this study are to evaluate the impact of a distressed environment on self-esteem in Jamaican adolescent girls and to assess the impact of behavioural patterns on self-esteem. Participants were African-Jamaicans ages 13-17 years from a Place of Safety (experimental group) and local high schools (control group). All participants completed the Rosenberg Self-Esteem Scale (RSES) while the experimental group also completed the Child Behaviour Checklist (CBCL). Participants in the experimental group were classified as either internalizers (depressed, anxious and somatizing), externalizers (exhibiting aggressive, disruptive behaviours), both (externalizers and internalizers) or other (reporting no or very few symptoms). It was hypothesized that the participants residing in the Place of Safety would report lower levels of self-esteem in comparison to the control group. Secondly, the 'both' group would report lower levels of self-esteem than any other group. Results indicated no differences in self-esteem scores; however, statistical significance was found between 'externalizers' and 'both' groups (p = 0.006). This study challenges the assumptions of self-esteem and its relationship to behavioural and emotional problems. PMID- 19565978 TI - Health determinants: using secondary data to model predictors of well-being of Jamaicans. AB - OBJECTIVE: During 1880-1882, life expectancy for Jamaican males was 37.02 years and 39.80 for their female counterparts and 100 years later, the figures had increased to 69.03 for males and 72.37 for females. Despite the achievements in increased life expectancies of the general populace and the postponement of death, non-communicable diseases are on the rise. Hence, this means that prolonged life does not signify better quality life. Thus, this study seeks to examine the quality of life of Jamaicans by broadening the measure of well-being from the biomedical to the biopsychosocial and ecological model METHOD: Secondary data were used for this study. The sample was a nationally representative one collected by the Statistical Institute of Jamaica and the Planning Institute of Jamaica in 2002. The total sample is 25,018 respondents of which the model used 1147. Data were stored and analysed using the Statistical Packages for the Social Sciences (SPSS). Multivariate regression was used to test the general hypothesis that well-being is a function of psychosocial, biological, environmental and demographic variables. RESULTS: The model explains 39.3 percentage of the variance in well-being (adjusted r2). Among those 10, the 5 most significant determinants of well-being in descending order are average number of persons per room (beta = -0.254, p < 0.001), area of residence (1 = KMA) (beta = -0.223, p < 0.001), area of residence [1 = Other Towns] (beta = -0.209, p < 0.001) and age of respondents (beta = -0.207, p < 0.001). These five variables accounted for 27.2 percentage of the model, with average occupancy and area of residence (being KMA) accounting for 7 per cent each. CONCLUSION: This study has shown that well-being is indeed a multidimensional concept involving psychosocial, environmental and demographic variables. PMID- 19565979 TI - Management of obstetric anal sphincter injuries at the University Hospital of the West Indies. AB - Up to 6% of women sustain severe perineal lacerations that involve the anal sphincters during vaginal delivery. When they occur obstetric anal sphincter injuries (OASI) may be accompanied by significant morbidity. Therefore, it is important to detect these injuries promptly and for experienced staff to perform sound repair. This report retrospectively assesses a series of seven women with OASI who were managed at a tertiary level hospital in Jamaica over a period of 28 months. Unfavourable details of management that may have adversely affected outcomes were sought from the various cases treated The incidence of OASI was low (0.2%). There were five third degree and two fourth degree lacerations. After these injuries were repaired, three patients (43%) experienced morbidity such as chronic pelvic pain (43%), anal incontinence (29%), dyspareunia (23%) and recto vaginal fistulae (14%). In order to improve the outcomes at this institution, several aspects of current care can be improved. Operative repair of these injuries should be delayed until senior staff is available to supervise OASI repair. Both methods of sphincter repair are reasonable options but the use of rapidly absorbable sutures is not appropriate. Finally, prophylaxis against wound infections can be achieved by administering a single dose of intravenous second or third generation cephalosporin at the time of induction of anaesthesia. PMID- 19565980 TI - Laparoscopic Heller's myotomy for treatment ofachalasia in the Caribbean. AB - The aim of this study is to examine the effectiveness with which Heller's myotomy can be performed laparoscopically in a Caribbean setting as a treatment for achalasia. Sixteen consecutive patients treated by the same surgeon were studied. Postoperative questionnaires that assessed symptomatology, complications and overall patient satisfaction with the procedure were utilized. The mean length of myotomy was 6 cm and none was converted to an open procedure. The mean age of the eleven female and five male patients was 38.4 years. The mean duration of follow up was 16.7 months. Symptoms showed an overall 71.2% improvement postoperatively (p < 0.001). Fifteen patients reported being satisfied with their operation with only one being dissatisfied Intra-operative complications occurred in three patients. Estimated blood-loss (EBL) was minimal in each case. Intra- and postoperative mortality was zero. Laparoscopic Heller's myotomy can be effectively performed in local settings and should be considered as an alternative to the open approach in the treatment of all surgically-fit achalasia patients. PMID- 19565981 TI - Donor site morbidity following iliac crest bone graft. AB - OBJECTIVES: To evaluate the complications of harvesting autogenous bone from the iliac crest. METHODS: A retrospective review of patients undergoing iliac crest bone grafting at the University Hospital of the West Indies, during the period 2000-2004, was performed. One hundred and three patients were identified. Thirty two patients were successfully contacted and 30 completed the questionnaire. There were 18 males (60%) and 12 females (40%). Their ages ranged from 13 years to 80 years (average 45.6 years). RESULTS: Of the 30 patients, 22 (73.3%) had complications. Fourteen (46.6%) patients had temporary pain; five (16.6%) had chronic pain. Two (6.6%) changed position of clothing due to discomfort at the graft site; five (16.6%) experienced difficulty walking, one reported itching of the scar one had altered sensation and one was unhappy with the scar. Fourteen patients (46.6%) had minor complications and eight patients (26.6%) had major complications. CONCLUSION: Autogenous iliac crest bone grafting is associated with significant complications. PMID- 19565982 TI - Chronic myeloid leukaemia at the University Hospital of the West Indies: a 17 year review. AB - OBJECTIVE: To determine the presenting features and evolution of patients diagnosed with chronic myeloid leukaemia between 1983 and 1999 at the University Hospital of the West Indies. METHODS: Forty-one records were retrospectively analyzed for the patients' demographics, reasons for referral, clinical features, laboratory investigations and the time to blast transformation and death. RESULTS: Seventy-one per cent were males and 29% were females. The male to female ratio was 2.4:1. The median age at presentation was 37 years (range 14-81 years). Seventy-eight per cent of the patients presented in the chronic phase. Weight loss and splenomegaly were the most frequent presenting features being seen in 54 and 83 per cent respectively. The median survival was 36 months. CONCLUSION: In this study, the clinical features and evolution were comparable to existing data. Improved accrual and routine Philadelphia chromosome testing would provide a more accurate reflection of the status of CML in our population. PMID- 19565983 TI - Cotrimoxazole resistance in Streptococcus pneumoniae isolated from sputum of HIV positive patients. AB - The prevalence and cotrimoxazole susceptibility of Streptococcus pneumoniae isolated from sputum of 100 HIV-positive patients attending the Nigeria Institute of Medical Research clinic was investigated using standard microbiological methods. Eleven of the sputum specimens grew Streptococcus pneumoniae. Antimicrobial susceptibility test showed that all the isolates were sensitive to amoxicillin, augmentin, erythromycin and chloramphenicol but were resistant to cotrimoxazole. Continuous surveillance of S pneumoniae in sputum samples of HIV positive subjects in this environment is necessary in order to regulate treatment regimen, considering that cotrimoxazole is the drug recommended by WHO for respiratory infections in HIV patients. PMID- 19565984 TI - Single photon emission computerized tomography in the evaluation of the osteoblastic activities of a new bone regeneration technique: analysis of 12 mandibular sites in six experimental pigs. AB - OBJECTIVES: To utilize single photon emission computerized tomography performed in sequence to determine the osseo-integrating capabilities and osteoblastic activities of a new bone regeneration technique called the membrane--sandwich technique (Ogunsalu sandwich bone regenerating technique) and to compare the quality and quantity of bone formed by this bone regeneration unit to bone regeneration obtained by using the same particulate bone grafting material covered with interceed (another type of bio-resorbable membrane). DESIGN AND METHOD: Single photon emission computerized tomography bone imaging was performed in sequence on the mandible of a total of 6 pigs on both the right and left side (total of 12 sites) at two and a half hours following the injection of technetium 99m methylene diphosphate. Imaging was performed using a Siemen Orbitar II gamma camera. The projection data was acquired in a 128 x 128 matrix over 180 arc and SPECT reconstruction was performed using a filtered back projector method with a Shepp-Logan Hanning filter and a cut-off frequency of 0.4. The surgical defect on one side of the jaw was treated with the sandwich unit with Bio-oss particulate bone within it, while the other side contained the same quantity of Bio-oss as in the left side but just covered with interceed membrane. The osteoblastic uptake on the side with the classical sandwich was compared to the side with the particulate bone covered with interceed membrane for dynamic physiological activities. The average activity for both sides was calculated and compared. RESULT: For all the 12 sites, osteoblastic activities were recorded and indicated that vascularized bone was formed at all the experimental sites. Autogenous bone graft was confirmed to be superior to xenograft using this sandwich technique. Furthermore, the osteoblastic activities on the sandwich side were seen to be more when compared with the control side (Interceed side). CONCLUSION: The Ogunsalu sandwich bone regeneration technique has been successfully evaluated with SPECT which shows osteoblastic activity with formation of vascularized bone which integrates with the surrounding bone. PMID- 19565985 TI - Simultaneous spontaneous bilateral pneumothoraces in an asthmatic. AB - The occurrence of simultaneous spontaneous bilateral pneumothoraces is a very rare event. We present a case of a 14-year old asthmatic female patient who presented to the emergency room for routine treatment. While receiving nebulizations, she suddenly developed supraclavicular fullness with crepitus. Further examination revealed a clinical diagnosis of bilateral pneumothoraces. Although this phenomenon is more commonly associated with patients on mechanical ventilation, this case illustrates that physicians must be cognizant of this unique presentation in order to initiate early and aggressive lifesaving therapy. With rapid bilateral needle thoracocentesis followed by placement of bilateral thoracostomy tubes, the patient recovered well. In this report, we also attempt to briefly review the possible pathophysiology of this form of spontaneous pneumothorax. PMID- 19565986 TI - Molecular analysis and implications of neurovirulent circulating vaccine-derived poliovirus in Jamaica. A case report and review of literature. AB - As the goal to eradicate wild polio virus (WPV) is approached, outbreaks associated with vaccine derived polioviruses (VDPV) with neurovirulent properties have emerged. The relevance for the spread of infection by nonparalytic cVDPV cases, with mutations associated with neurovirulence, is discussed with reference to the molecular analysis of a VDPV isolated from a Jamaican child who presented with aseptic meningitis. Potential risks to the Jamaican community resulting from circulation of cVDPV and critical factors defined by the World Health Organization (WHO) in the global eradication of Polio are analyzed in the context of immunization coverage, and the need to stop all Oral Polio Vaccine (OPV) use once wild polioviruses (WPVs) have been eradicated. PMID- 19565987 TI - Emergency Whipple's at a rural hospital in Jamaica. PMID- 19565988 TI - The role of surgeon specialization in improving patient outcomes: a challenge for the Caribbean. AB - Progressive surgeon specialization has been shown to result in improved patient outcomes for many surgical procedures. This has been demonstrated in improved survival following cancer surgery and improved operative morbidity and mortality for other procedures. Concentrating complex surgical cases in centres where case volume and expertise exist should result in better overall surgical care delivery. PMID- 19565989 TI - Chromosomally mediated antibiotic resistance in non-typhoidal Salmonellae isolated from HIV patients in Lagos. PMID- 19565990 TI - A new challenge: depression is a significant problem among university students. PMID- 19565991 TI - Effect of a government funded medication programme on paediatric asthma hospital admissions in Antigua and Barbuda. AB - OBJECTIVE: To assess the effect of a government funded asthma medication programme on paediatric (age < or = 12 years) asthma hospital admissions in Antigua and Barbuda. METHODS: A retrospective review of all hospital admissions for asthma in children was performed for the six years before and six years after a Medical Benefits Scheme (MBS) programme was established in 1997 to provide asthma medications at no out-of-pocket cost. Holberton Hospital records (1992 to 2003) which include all paediatric asthma admissions in Antigua and Barbuda, were reviewed RESULTS: Paediatric admissions for asthma fell from mean +/- standard deviation of 77.0 +/- 24.8 per year before the MBS programme was started to 48.0 +/- 17.1 per year (p < 0.05) after the MBS programme was started. The number of multiple admissions fell from 18.7 +/- 2.7 to 9.5 +/- 4.8 (p < 0.005) and the number of children admitted multiple times per year fell from 7.8 +/- 1.9 to 4.7 +/- 2.5 (p < 0.05). The number of children aged four to nine years admitted with asthma fell from 7.8 per 1000 annually during 1992 to 1997 to 4.4 per 1000 per year during 1998 to 2003. CONCLUSIONS: The government funded MBS programme for asthma medication has resulted in a 38% decrease in hospital admissions for paediatric asthma over a six-year period. The benefits of a similar programme in other developing countries should be considered. PMID- 19565992 TI - Laparoscopic Nissen fundoplication for improved gastrointestinal symptoms and quality of life. AB - OBJECTIVE: The purpose of this study was to establish baseline data for a procedure that has yet to be defined within a Caribbean population. Using a specifically designed postoperative questionnaire, symptomatology and quality of life were assessed before and after laparoscopic Nissen fundoplication. METHODS: Twenty-three consecutive patients of the same surgeon were identified. The questionnaires measured the symptoms and social and emotional functioning, assigning each a score for comparative purposes. The cumulative score was considered the ultimate index of overall quality of life. Wilcoxon matched pairs test was used to analyse the data. RESULTS: The mean age of the patients was 47.5 years. There were 21females and 2 males. Heartburn (78.3%) and regurgitation (60.8%) were the main symptoms. Postoperatively, these decreased to 17.3% and 4.3% respectively, with significant declines in other complaints such as dysphagia and nausea. The social and emotional functioning score had an average of 13.2 preoperatively; this increased to 18.3, postoperatively, out of a possible maximum of 20. None of the procedures had to be converted to open laparotomy and mean operating time was 108 minutes. Mean follow-up time was 9.7 months. CONCLUSION: Laparoscopic Nissen fundoplication does provide significant improvement in overall quality of life among patients with gastrointestinal symptoms and can be performed effectively within a Caribbean setting. PMID- 19565993 TI - Reliability of the evaluation of students' answers to essay-type questions. AB - INTRODUCTION: This paper seeks to quantify the reliability of the assessment of students' answers to essay-type questions, in an attempt to define the role of such questions in University examinations. METHODS: The marks awarded for essay type questions during three consecutive final undergraduate examinations in surgery were analyzed. The mean scores, 95% confidence intervals and the standard error of the mean were calculated to determine the distribution of the marks. Statistical analysis was used to determine the correlation of the marks awarded for the same answer by different markers and deduce the dependability of this method of testing. RESULTS: The marks awarded to 233 answer papers were available for analysis. The marks awarded by each pair of examiners for student answers to individual questions coincided on only 46.3% of occasions, but varied within just +/- 5% on 90.7% of occasions. Use of the kappa index to determine the agreement between markers produced a value of just 0.385, well short of the ideal of 1.0. Assessment of the overall reliability of this type of examination by Cronbach's reliability coefficent gave a value of 0.672. CONCLUSION: There was a significant variation among markers in the evaluation of answers to essay-type questions. However; the overall test reliability was acceptable enough to justify continuation of this type of assessment as a supplement to other methods. PMID- 19565994 TI - Glycated haemoglobin A1c measurement in stored whole blood sample is reliable for clinical use. AB - Glycated haemoglobin A1c (HbA1c) gives an integrated plasma glycaemia for the previous 2-3 months and its measurement is central in the management of diabetic patients. However in many developing countries because kits/regents or expertise for HbA1c measurement are not always available and the test must be conducted on fresh whole blood samples, HbA1c tests are not routinely performed Thus, this study aimed to determine if the degradation products from whole blood sample storage are significant enough to compromise the diagnostic value of HbA1c measurements. Two hundred and thirty-one fresh whole blood samples with pre determined HbA1c values were stored at between 2-8 degrees C and using boronate affinity immunoassay technique, HbA1c values were then measured in the same whole blood samples after 20 days of storage. The results showed that there were no significant differences in the mean values of the initial HbA1c measurement and the values obtained after storage (7.5 +/- 2.0 vs. 7.5 +/- 2.1, p > 0.05) and this was irrespective of gender. Furthermore, irrespective of gender there were significant correlations between the HbA1c values measured in fresh whole blood samples and values obtained after storage (r = 0.83, p < 0.01). Therefore, based on these findings and other previous reports, the effect of storage degradation product was not significant enough to compromise the clinical or research use of HbA1c test results from stored whole blood samples. However, we recommend that diagnostic laboratories should evaluate their HbA1c measurement techniques for HbA1c determination in stored whole blood samples. Any persistent upward or downward bias in stored whole blood samples should be reported to guide the physician in interpreting HbA1c results from stored whole blood samples from that laboratory and/or technique. PMID- 19565995 TI - Factors associated with depression in students at The University of the West Indies, Mona, Jamaica. AB - OBJECTIVE: This project examines the factors associated with depression in students attending the University of the West Indies, Mona Campus. METHOD: Students enrolled in the Foundation courses during the first and second semesters of the 2005/2006 academic year were administered the Brief Screen for Depression as well as a demographic questionnaire as part of a larger study. RESULTS: A wide cross-section of the university population was sampled (n = 690; 252 from semester one, 438 from semester two; 77% females, 23% males; age 16-62 years, median = 20 years, mean = 23.4 years +/- 7.4). Nearly 40% of students scored in the clinically depressed range. Students in the December wave of data collection had higher depression scores than those in the January wave. Consistent with international research, females reported significantly higher levels of depressive symptoms. Married students reported significantly lower depression scores than students in visiting relationships. Students who were combining employment and school reported lower depression scores than those who were not employed. Maternal education significantly influenced students' levels of depression such that students whose mothers had university or other tertiary education had lower depression scores while those whose mothers had primary or lower education had the highest depression scores. Students with a chronic condition or a disability scored higher than those without such problems on all three measures of depression. CONCLUSION: Depression may be a significant problem in students at The University of the West Indies, Mona campus. PMID- 19565996 TI - Sociocultural deterrents to mammographic screening in Jamaica. AB - OBJECTIVE: Less than five per cent of eligible Jamaican women had mammograms in 2003. The sociocultural determinants and the perceptual barriers modulating screening behaviour in Jamaican women are unclear. We sought to investigate sociocultural effects, in particular knowledge and fear of the procedure on mammographic screening behaviour in Jamaican women. METHOD: One hundred and forty seven women attending the breast imaging units at the University Hospital of the West Indies and 127 attending Radiology West were interviewed to determine the factors relating to participation in mammographic screening. Knowledge level, deterring factors as well as the experience during mammography were recorded. RESULTS: The mean age +/- SD of participants was 51 +/- 10.4 years. Eighty-eight of the 274 women (32%) were having a mammogram for the first time. Of these, the major determinants of the mammographic experience were the expectation that the procedure would be painful (OR = 0.08, p < 0.001) and the pain intensity (OR = 1.4, p < 0.030) experienced during mammography. There were 188 women who had repeat mammograms. Seventy-five of these women had delayed mammography for greater than one year. There was a significant association between being late for mammography and the perception that a doctor's referral was necessary (p < 0.001). The factors associated with improved mammographic experience were pain intensity (OR = 0.84, p < 0.04), interval status of previous mammography (OR = 2.24, p = 0.059) and knowing someone with breast cancer (OR = 0.35, p < 0.04). Although 97% of all women found mammography painful, only seven (2.5%) said pain would prevent a repeat mammogram. CONCLUSIONS: Fear pain during mammography, subjective indifference, inertia and reliance on physician referrals were identified as barriers to complying with mammographic screening guidelines. PMID- 19565997 TI - Diseases and medical disabilities of enslaved Barbadians, from the seventeenth century to around 1838 part II. AB - The disease environment, health problems and causes of mortality of enslaved Barbadians are described. Data are derived mainly from documentary sources; also included are bio-archaeological data from analyses of skeletons recovered from Newton Plantation cemetery. Major topics include infectious diseases transmitted from person to person, as well as those contracted through water soil, and other environmental contaminations, and diseases transmitted by insects, parasites and other animals; nutritional diseases, including protein energy malnutrition, vitamin deficiencies, anaemia, and geophagy or "dirt eating"; dental pathologies, lead poisoning, alcoholism, traumas, and other disorders, including psychogenic death or illness caused by beliefs in witchcraft or sorcery. PMID- 19565998 TI - The effects of 201Tl myocardial perfusion scintigraphy studies on oxidative damage in patients. AB - The aim of this study was to investigate gamma radiation-induced oxidative damage in erythrocytes after 201Tl myocardial perfusion scintigraphy. Twenty patients (8 women and 12 men) who performed 201Tl myocardial perfusion scintigraphy were included in this study. The blood samples were taken from patients just before, 1 hour after and three hours after injection of the radiopharmaceutical. Malondialdehyde (MDA) and antioxidant enzymes such as glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (CAT) levels were measured to evaluate the gamma radiation induced oxidative damage. The enzyme activities of SOD, GPX and CAT were decreased 1 hour after (p = 0.042, p = 0.697 and p = 0.653 respectively) and 3 hours after (p = 0.003, p = 0.573 and p = 0.002 respectively) injection of the radiopharmaceutical. Malondialdehyde levels were increased 1 hour after (p = 0.10) and 3 hours after (p = 0.47) injection of the radiopharmaceutical. In this study, we found that radiation due to 201Tl myocardial perfusion scintigraphy decreased the erythrocyte antioxidant levels and increased MDA levels. PMID- 19565999 TI - Probing the specifics of substrate binding for cytochrome c oxidase: a computer assisted approach. AB - A deficiency of cytochrome c oxidase (COX) is associated with a number of diseases but details of the enzyme's mechanism of action especially the interaction with its substrate, ferrocytochrome c, remain unclear. It is known that the transfer of electrons from ferrocytochrome c to COX is facilitated by the formation of enzyme-substrate (ES) complexes which are stabilized by intermolecular salt bridges, however the identity of residues participating in the salt bridges remains obscure. Using the published structures of the two proteins, computer simulations were employed to model their interactions and to attempt to identify residues that participate in intermolecular salt bridges. The simulation process was guided in the main by cross-linking studies which proposed that Lys-13 of cytochrome c is paired with Asp-158 of COX. The initial enzyme substrate complex, created by computer assisted manipulation of the two structures exhibited three salt bridges; following the application of energy minimization procedures, the number of salt bridges increased to seven and there were twenty-four intermolecular hydrogen bonds. The salt bridges emanated from: Glu-119 and Asp-221 of subunit I; Glu-114, Asp-115 and Asp-158 of subunit II and Asp-73 and Glu-78 of subunit VIb. These were paired with Lys-87, 8, 25, 27, 13, 22 and 100 respectively of cytochrome c. These results suggest that subunits I, II and VIb play direct roles in substrate binding. The results also suggest that hydrogen bonds contribute significantly to the stability of the ES-complex. PMID- 19566000 TI - Retroperitoneal pararenal Castleman's disease. AB - A 51-year old male patient with a three-month history of constant and dull left flank pain was investigated by ultrasonography, computed tomography (CT) scan and magnetic resonance imaging (MRI) of the abdomen, which disclosed a 8 x 7 x 6 cm retroperitoneal pararenal mass with heterogeneous imaging characteristics and bright enhancement following intravenous contrast injection. Based on the hypervascularity of the mass and the lack of specific signs in the imaging investigation, lymphoma, sarcoma or vascular tumour were considered as probable, diagnoses and the patient underwent an exploratory laparotomy. The histologic examination of the surgically resected specimen disclosed "a hyaline type of Castleman's disease". Further evaluation of the patient with antibody testing for HIV 1 and 2, as well as viral load by PCR for Herpes Virus-8 (HHV-8) were negative. Bone marrow aspiration, biopsy and immunophenotypic study did not disclose any evidence of lymphoma. Molecular study of the bone marrow for immunoglobulin heavy chain rearrangement showed a polyclonal pattern; serum protein electrophoresis did not show any evidence of hypergammaglobulinaemia and serum immunofixation electrophoresis did not show any monoclonal protein. A diagnosis of localized-unicentric type of Castleman's disease was made. Castleman's Disease should be included in the differential diagnosis of any solitary, heterogeneous and hypervascular retroperitoneal mass. Discovery of Castleman's disease at any area of the body should be followed by a thorough imaging and laboratory work-up in order to exclude the multicentric type of the disease and the co-existence of lymphoma. PMID- 19566001 TI - Manchineel poisoning bradyarrhythmia: a possible association. PMID- 19566002 TI - Spontaneous resorption of exudative retinal detachments associated with ischaemic central retinal vein occlusion. PMID- 19566003 TI - Systemic lupus erythematosus and hyper-eosinophilic syndrome: an unusual association. AB - A 34-year old woman with dizziness, headache and both upper and lower extremities weakness was admitted to hospital. She had a history of photosensitivity but no asthma or allergy. On physical examination, malar rash and livedo reticularis were noted. White blood cell count was 18500/microL with 7585 eosinophils (41%). She also had positive antinuclear antibody (ANA), anti-double stranded DNA antibody (anti-ds-DNA antibody) and anticardiolipin antibody (aCL antibody). Echocardiography revealed left and right ventricular obliteration with fibromatous biventricular endothelial thickening. Brain MRI showed changes in favour of white matter ischaemia and lacunar infarction. Hypereosinophilic syndrome (HES) and systemic lupus erythematosus may be considered to have occurred concurrently in this patient. PMID- 19566004 TI - IgA nephropathy in the Caribbean case reports. AB - IgA nephropathy can be considered the most common cause of primary glomerulopathy in developed countries. There has been no report of cases of IgA nephropathy from Caribbean countries. The authors report five cases of IgA nephropathy from Trinidad and Tobago, and Guyana, diagnosed from biopsy studies. No cases were of African origin and some did not have the typical presentation associated with IgA nephropathy. Caribbean nephrologists are reminded that this entity can be seen in Caribbean patients and can only be diagnosed through immunofluorescence staining of renal biopsy specimen. This diagnosis is required for the proper management of patients with glomerular disease, particularly when there may be progression to end stage renal failure as can occur in up to twenty per cent of patients with IgA nephropathy. Accurate diagnosis is important, since disease recurrence can be seen in the transplanted kidney, but this does not often lead to graft failure. PMID- 19566005 TI - Tracheobronchial stenting for management of bronchopleural fistula. PMID- 19566006 TI - Specific measures to reduce walk-in deliveries in Japan. PMID- 19566007 TI - The current state of gastric cancer mortality in Ishikawa and Tokushima Prefectures and establishment of cancer prevention measures. PMID- 19566008 TI - Health information systems in the Caribbean: time for attention. PMID- 19566009 TI - Protective effect of Carica papaya L leaf extract against alcohol induced acute gastric damage and blood oxidative stress in rats. AB - The effects of Carica papaya leaf (CPL) aqueous extract on alcohol induced acute gastric damage and the immediate blood oxidative stress level were studied in rats. The results showed that gastric ulcer index was significantly reduced in rats pretreated with CPL extract as compared with alcohol treated controls. The in vitro studies using 2,2-Diphenyl-1-Picryl-Hydrazyl (DPPH) assay showed strong antioxidant nature of CPL extract. Biochemical analysis indicated that the acute alcohol induced damage is reflected in the alterations of blood oxidative indices and CPL extract offered some protection with reduction in plasma lipid peroxidation level and increased erythrocyte glutathione peroxidase activity. Carica papaya leaf may potentially serve as a good therapeutic agent for protection against gastric ulcer and oxidative stress. PMID- 19566010 TI - The in vitro anti-denaturation effects induced by natural products and non steroidal compounds in heat treated (immunogenic) bovine serum albumin is proposed as a screening assay for the detection of anti-inflammatory compounds, without the use of animals, in the early stages of the drug discovery process. AB - There are emerging ethical issues with regards to the use of animals in the early stages of drug discovery for anti-inflammatory and degenerative diseases from natural products using the activity-directed isolation pathways when many compounds (eg > 100) are present in the crude extract or fraction and are to be tested The above-mentioned is the main reason for proposing the use of the in vitro anti-denaturation (stabilization) effects of heat treated (immunogenic) bovine serum albumin (BSA) as an assay. Current methods used for detecting and isolating a wide range of anti-inflammatory compounds in the early stages of the drug discovery process utilize a large number of animals. When BSA is heated and is undergoing denaturation, it expresses antigens associated to Type III hypersensitive reaction and which are related to diseases such as serum sickness, glomerulonephritis, rheumatoid arthritis and systemic lupus erythematosus. Thus, the assay that is being proposed should be applicable to the discovery of drugs for treating the above mentioned diseases and others, once the compounds stabilize the denaturation process. PMID- 19566011 TI - The initiation of coronary angioplasty and stenting in a single outpatient centre in Barbados. AB - Percutaneous transluminal coronary angioplasty (PTCA) is a novel procedure to Barbadian healthcare. Only one centre in Barbados provides PTCA and stenting. This is a retrospective study aimed at describing the initial results of coronary angioplasty and stenting in the first 48 patients at the Carib-American Heart Centre and exploring the feasibility and safety of coronary angioplasty and stenting in Barbados. Forty-eight patients underwent PTCA during the period March 2002 to June 2004 inclusive, with or without intracoronary stenting. Most (64.6%) of the patients were male and 43.7% were diabetic. The most common vessels involved were the left anterior descending coronary artery (LAD) and the right coronary artery (RCA). In one patient, attempted stenting was unsuccessful but PTCA reduced stenosis. One patient had previous PTCA and stenting of the LAD and two patients had coronary artery bypass grafting (CABG) prior to the procedure. Twventy-one per cent of the patients treated had severe triple vessel disease. There were no cases of re-stenosis or acute vessel closure, during or immediately following the procedure that required emergency PTCA or CABG All patients were discharged within 24 hours of the procedure. Procedural success was 100%. In conclusion, outpatient PTCA and stenting is safe and feasible in the Barbadian population. Coronary artery bypass grafting is still the procedure of choice for treating coronary artery disease (CAD) involving the left main coronary artery but PTCA is indicated in some cases of severe triple vessel disease. PMID- 19566012 TI - Clinical determinants of increased left ventricular mass on echocardiogram in medically treated Afro-Caribbean hypertensive patients. AB - Increased left ventricular mass (LVM) on echocardiogram is an independent risk factor for cardiac complications from hypertension. It is associated with a four fold increase in untoward cardiac events when present. Data were reviewed for 100 treated hypertensive Afro-Caribbean patients, aged 29 to 65 years, recruited from village health clinics. Age, gender, height, weight, systolic and diastolic blood pressure, echocardiogram (ECHO) and drug history were recorded for each patient. The best single predictor of increased LVM was blood pressure with systolic (163 vs 152 mmHg, p < 0.01) and diastolic blood pressure (105 vs 98, p < 0.01) being significantly higher in patients with increased LVM. Systolic blood pressure over 150 mmHg was associated with increased LVM in 64% vs 44% below 150 mmHg (p < 0.10). Diastolic blood pressure over 95 mmHg was associated with increased LVM in 63% vs 36% below 95 mmHg (p < 0.02). BMI showed a trend (31.1 vs 29.7 kg/m2, p < 0.20) toward higher values in patients with increased LVM. BMI above 28 kg/m2 was associated with increased LVM in 61% vs 44% below that value (p < 0.15). Females more than males had increased LVM (61% vs 44%, p = 0.30) possibly due to higher BMI (31 vs 29.4 kg/m2) and higher systolic blood pressure (160 vs 155 mmHg). Age (48.3 vs 46.5 years, p = 0.30) and years of hypertension (8.6 vs 7.3 years, p = 0.33) were not significantly different between the two groups. Drug treatment was reported in 90% (69% monotherapy, 27% > one drug, 4% > 2 drugs) and no drug was associated with significant difference in LVM compared to others. Only 15% of treated hypertensive patients had systolic blood pressure below 140 mmHg and 8% had diastolic blood pressure below 90 mmHg. The major determinant of increased LVM in this group of Afro-Caribbean hypertensive patients appears to be poorly controlled hypertension with obesity being a possible contributing factor PMID- 19566013 TI - Echocardiographic findings in a contemporary Afro-Caribbean Population referred for evaluation of unexplained syncope. AB - Echocardiographic findings were reviewed for 106 patients (mean age 41.3 +/- 23.0 years, range 3 to 90 years, 61% female) referred for evaluation of unexplained syncope. Abnormal echocardiographic findings were seen in 36/106 (34%) patients, of which 12/106 (11%) may have an abnormality that contributed to symptoms. Abnormal echocardiographic findings (64 vs 6%, p < 0.01) and those possibly causing syncope (22 vs 0%, p < 0.05) were significantly more likely in the oldest tercile of patients compared with the youngest. No patient less than 35 years old had a possibly diagnostic abnormality. PMID- 19566014 TI - Increasing incidence of admissions to a general hospital for deliberate self-harm in Trinidad. AB - BACKGROUND: Deliberate self-harm and suicidal behaviour have become an increasingly common form of morbidity in the developing world Suicidal behaviour is a major public health problem in Trinidad as the country has the second highest completed suicide rate in the English-speaking Caribbean. The objectives of this study were to determine the epidemiology of attempted suicide and self harm at a specified site (the Port-of-Spain General Hospital) and compare it to previous studies done at the same site. SUBJECTS AND METHODS: This was investigated through a review of one years' admissions to the Port-of-Spain General Hospital for suicidal behaviour. Incidence was compared with a previous study completed at this site and reported in 1974. Comparison of the demographic characteristics of the sample with that of the previous study was also undertaken using chi-square analysis and significance testing through the use of t tests. RESULTS: A total of 368 referrals were made for attempted suicide or deliberate self-harm over the period indicating a fourfold increase in the incidence of this behaviour with a greater increase among males where the female to male ratio has declined from 4 to 1 to 2 to 1, p < 0.001; the mean age of males was 34 years compared to females 22 (p < 0.0001). The males were more likely to use violent means compared to females who were more likely to ingest tablets or bleach. Relationship difficulties were most commonly cited by both males and females as the reason for their attempt. There was a trend to greater propensity for this behaviour among Indo-Trinidadians as compared to Afro-Trinidadians in both females and males. CONCLUSIONS: Increasing numbers of men are engaging in self harm behaviour and are using more violent and physically harmful methods suggesting a greater degree of suicidal risk while women mainly engage in acts of ingestion with a much lower risk of death. The older mean age of these men suggests that their problems are presenting in middle adulthood while women are engaging in this behaviour in young adulthood. Suicidal behaviour or deliberate self-harm is a major public health problem in Trinidad. PMID- 19566015 TI - Depression among adolescents, aged 13-19 years, attending secondary schools in Trinidad: prevalence and associated factors. AB - OBJECTIVE: To determine the prevalence of depression and psychosocial factors associated with depression in secondary school students in Trinidad METHODS: This was a cross-sectional study of a stratified random sample of public secondary schools utilizing a modified pre-tested self-administered Beck Depression Inventory (BDI) to detect depression in students aged 13-19 years in Trinidad. RESULTS: In this study, 1290 students participated, a response rate of 79.6%; 43% were aged 13-15 years; 53.6% were Indo-Trinidadians; 82.5% were attending co educational schools and 70.6% lived with both parents. The prevalence of depression was 25.3% +/- 2.37%. Chi-square analysis revealed statistically significant associations between depression and the categories of age, gender, living arrangements and school type. Similar findings were observed for respondents who admitted to cigarette and alcohol use or to being afraid of or being injured by their parent (p < 0.05). Logistic regression indicated that females were 1.7 times as likely to be depressed when compared with males; respondents not living with both parents were 1.5 times as likely to be depressed as those who were. Respondents reporting that they were afraid of parents or of being injured by parents were three times as likely to be depressed as respondents who had not had those experiences. CONCLUSIONS: One out of every four secondary school students in Trinidad was found to have significant depression. There were strong associations between depression and age, gender, school type and family structure. This study identifies that many adolescents experience violence in the home and those who did were more likely to be depressed. PMID- 19566016 TI - Toward effective school-based substance abuse prevention "breaking the cycle" programme in Antigua and Barbuda. AB - The "Breaking the Cycle" programme, based on the Project Charlie programme, was developed for Antigua and Barbuda third grade students and was implemented in 2001. Aspects of the programme are compared with aspects recently proven effective in randomized studies in developed countries. The "Breaking the Cycle" programme includes life-skills training, teaches decision making skills, includes peer resistance training, uses trained teachers, interactive teaching methods, effective content and delivery, targets students prior to onset of drug use, teaches drug harm, teaches community values and is culturally sensitive, all aspects of successful programmes overseas. The cost of about $7 US per student would suggests cost-benefit effectiveness compared with overseas programmes. The "Breaking the Cycle" school-based drug and alcohol use prevention programme includes most aspects of evidence-based successful programmes overseas, appears cost effective and could serve as a model for programmes in the Caribbean region. PMID- 19566017 TI - Factors affecting completion of a 28-day inpatient substance abuse treatment programme at the University Hospital of the West Indies. AB - This study investigated factors leading to premature discharge from the residential substance abuse treatment programme at the University Hospital of the West Indies (UHWI). Two hundred and twenty-four patients, mean age 36 +/- 9.75 and age range of 16-66 years were studied The patients, 87.5% men, were admitted to the programme over the period July 1999 to June 2002. Sources of data, including a routine patient-monitoring form and patients' medical records, were reviewed. Data analysis, performed using the Statistical Package for Social Sciences (SPSS), included frequencies, cross-tabulations and analyses of variance (ANOYA). Crack cocaine was the drug of impact for 60% of patients, alcohol for 17% and marijuana for 8%. Treatment ended prematurely for 22.8% of persons whose reasons for leaving included signing out against medical advice or self-discharge (27.4%), absconding (19.6%), fighting (9.8%), drug use (5.9%) and other rule breaking, including sexual acting out (15.7%). Premature discharge was uninfluenced by gender, age, marital status, schooling, employment status or geographical location. There were significantly more premature discharges in persons admitted primarily for crack dependence versus those admitted primarily for alcoholism (p < 0.05). There were also significantly more premature discharges in persons with 0-10 years of using their most problematic substance when compared with 21-30 years of use (p < 0.05) but not when compared with 11-20 years or over 30 years of use (p > 0.05). The influence of duration of substance use requires further elucidation. PMID- 19566018 TI - Routine laboratory investigations in infants and children presenting with fever and seizures at the University Hospital of the West Indies. AB - A retrospective chart review of the case notes of all children aged 6 months to 8 years presenting with fever and seizures to the University Hospital of the West Indies (UHWI) between January 2000 and December 2004 was conducted. Descriptive analyses were performed. Fifty-nine children (median age 1.58 years, range 0.58 to 6.83 years) were entered into the study. The main laboratory abnormalities were metabolic acidosis (23%), anaemia (10%), leukocytosis (35%) and hypomagnesaemia (3%). These were not significantly associated with meningitis or an underlying bacterial infection. There were no significant episodes of hyponatraemia, hypocalcaemia or hypoglycaemia. Meningitis was uncommon and occurred in only two (3.4%) children both younger than 16 months of age and who had other abnormal clinical signs. This study demonstrated that routine performance of haematological and biochemical investigations in children presenting with seizures and fever were of limited value. Lumbar punctures in children older than age 18 months with no other abnormal clinical signs were also found to be of low yield Current American Academy of Paediatrics (AAP) recommendations that serum electrolytes, calcium, phosphate, magnesium, complete blood count and blood glucose should not be performed routinely in a child with a first simple febrile seizure can be safely applied to this study population. PMID- 19566019 TI - Dengue infection in patients presenting with neurological manifestations in a dengue endemic population. AB - The evaluation of the contribution of neurological dengue in suspected central nervous system (CNS) viral infections is essential to better understand the impact of neurological dengue on morbidity and mortality in dengue endemic regions such as Jamaica. For this study 401 cases of suspected viral CNS infections were investigated for evidence of dengue infection. The frequency of neurological dengue among these CNS cases was found to be 13.5% (54/401). Fifty three cases were confirmed serologically by haemagglutination inhibition assay (HI) and IgM antibody (ELISA) and the virus was isolated in one case only. Clinical manifestations among dengue positive CNS cases included encephalitis in 51.8% (28/54), meningitis in 33.3% (18/54), seizures in 11.1% (6/54) and acute flaccid paralysis/Guillain-Barre syndrome in 3.7% (2/54). The clinical diagnosis of dengue neurological infection corresponded with laboratory confirmation in 22.2% (12/54) of cases only. Deaths occurred in 3.7% (2/54) of cases and were associated with patients with dengue neurological infection. The high risk of dengue among patients with suspected viral CNS infections in this study supports the need for an increased index of suspicion of dengue in patients presenting with neurological manifestations in dengue endemic countries. PMID- 19566020 TI - Prevalence, risk factors and clinical features associated with intestinal parasitic infections in children from San Juan y Martinez, Pinar del Rio, Cuba. AB - A cross-sectional study was carried out in 200 children aged 5-15 years, to examine the presence of intestinal parasitic infections (IPIs) and to assess the risk factors and clinical features associated with them in children in San Juan y Martinez (SIM), Cuba. Three fresh faecal samples were collected from each child and were examined by direct wet mount, brine flotation, formalin-ether and Kato Katz techniques. Data relating to demography, source of drinking water, personal hygiene habits and clinical features were also collected Living in the rural area was significantly associated with the highest infection rates (p < 0.01). According to clinical features and laboratory examinations, children with abdominal pain were about four times more likely to have IPIs (OR 4.05, CI, 1.11, 13.18) especially soil-transmitted helminths (STH). We suggest that IPIs, especially STH, in SJM should be strongly suspected in children with abdominal pain from rural areas. Targeted and frequent interventions to control these infections are needed in this municipality. PMID- 19566021 TI - A comparative study of the quality and availability of health information used to facilitate cost burden analysis of diabetes and hypertension in the Caribbean. AB - OBJECTIVE: Non-communicable Diseases (NCDs) are leading threats to health and well-being in the Caribbean. A study was undertaken in the latter part of 2005 to compute the economic burden of diabetes mellitus and hypertension within the Caribbean Community and Common Market (CARICOM). This report critiques the quality and availability of health information which can be used to facilitate cost burden analysis of diabetes mellitus and hypertension. METHODS: A form was developed and disseminated to obtain epidemiological and health service utilization data. Subsequent visits were made to seven CARICOM member countries to collect the data. RESULTS: The results revealed (i) a number of deficiencies in the reliability and validity of the data received, in particular, those needed to facilitate the analysis of cost-specific complications such as ischaemic heart disease, cerebrovascular disease, chronic renal failure, hypertensive and diabetic retinopathy and peripheral circulatory complications; (ii) data management systems in hospitals were not linked to facilitate generation of cost effectiveness estimates and other information required to compare options for health investment; (iii) despite repeated attempts by regional governments to develop/strengthen Health Information Systems within the Caribbean, sustainability has been significantly hampered by human, material and financial resource constraints and ongoing monitoring and evaluation is generally poor. CONCLUSION: There are deficiencies in the quality and availability of health information to facilitate cost burden analysis of hypertension and diabetes mellitus in the Caribbean Strong commitment from CARICOM governments will be necessary to address these concerns if economic evaluations are to be undertaken more frequently as part of the effort to reduce the morbidity and mortality from these diseases. PMID- 19566022 TI - Interstitial cells of Cajal and intestinal function in Trinidadian children. AB - OBJECTIVE: This study was carried out to compare the density of the interstitial cells of Cajal (ICCs) in the bowel wall of children with Hirschsprung's disease (HD), anorectal malformations (ARM) and normal controls in Trinidad and Tobago. SUBJECTS AND METHOD: Segments of bowel wall excised from eight children with HD, three controls and two children with ARM were immunostained with c-Kit primary antibody. Cells with features of ICCs were counted. RESULTS: All three controls and the two children with ARM had dense distribution of ICCs. Most children (6/8; 75%) with HD had markedly reduced counts in aganglionic bowel. Two (25%) also had a decrease in ganglionic bowel. Possible influences were patient age and gender and the level of bowel sectioned. CONCLUSION: Analysis of this sample suggests that immunostaining for c-Kit positive cells might be a useful screening test in the assessment of bowel motility disorders. The possible effects of age, gender and the level of bowel sampled await determination. PMID- 19566023 TI - Prevalence of Chlamydia trachomatis/Neisseria gonorrhoeae and human papilloma virus among women-at risk in the Aegian region of Turkey, and their knowledge about HIV/AIDS. AB - BACKGROUND: The aim of this study was to determine the prevalence of selected sexually transmitted diseases (STDs) and the level of knowledge and attitudes regarding HIV/AIDS among Turkish brothel based sex-workers (SWs). SUBJECTS AND METHODS: A pre-designed questionnaire was administered to 199 SWs to obtain their sexual behaviours and their level of knowledge of HIV/AIDS. The specimens collected for C trachomatis/N gonorrhoeae and human papillomavirus (HPV) were tested using Gen-Probe PACE 2 and HPV-screening assays, respectively. RESULT: Aproximately sixty-seven per cent of the SWs knew that condoms afforded protection against HIV/AIDS and 62% reported continued use of condoms. Although most of the SWs had heard about HIV/AIDS, thorough knowledge of transmission and prevention was lacking. The overall estimated rates for C trachomatis/N gonorrhoeae and HPV were 18.6% and 9.7%, respectively. CONCLUSION: There is a need for further studies to generate more data on the prevalence of STDs and the knowledge of STDs in this population. PMID- 19566024 TI - An abnormal variation of the brachial plexus with potential clinical significance. AB - In this case of a male adult Caucasian cadaver, variations were noted involving the location of the CS ventral ramus descending anterior and superolateral to the anterior scalene muscle. An unusual communicating branch was discovered that originated from C5 at the midpoint of the anterior scalene muscle, two centimeters proximal to Erb's point, to communicate with C6 and another communicating branch originating from C6 to C7. Awareness of the possibility of this variation is of great importance during certain surgical procedures. Comparison of this case with the literature is discussed. PMID- 19566025 TI - Congenital Cutis laxa associated with growth retardation. AB - Congenital cutis laxa is a rare, clinically and genetically heterogeneous group of inherited disorders. It is characterized by degenerative changes in elastic fibres and manifests with skin laxity. Here we presented a six-month old boy with congenital cutis laxa associated with growth retardation. We reveal ultrastructural findings and discussed the differential diagnosis. PMID- 19566026 TI - Assessment of alveolar bone levels from bitewing, periapical and panoramic radiographs in periodontitis patients. AB - The aim of this study was to determine the relationship between loss of radiographic crestal bone height in panoramic, bitewing and periapical radiography and to probe the attachment loss after periodontal treatment. Radiographic and probing measurements were made at baseline and after one year The population in the study consisted of 21 individuals, 13 females and 8 males, ages 18-59 [mean 35.7] years. A total of 42 interproximal intra-bony defects of 21 mandibular first molar teeth, treatments of which were planned by subgingival curettage procedures, were selected from among 21 subjects who had signed consent forms. Measurements of the distance between the cemento-enamel junction and the alveolar crest were compared with probing crestal bone level and radiographic measurements before and one year after the procedures. The results of probing pocket depths level before and one year after subgingival curettage were different with the mean average being 1.18 +/- 1.51 mm. These changes were statistically significant (p < 0.05). Bitewing radiography showed the highest accuracy among radiographic methods in the assessment of the crestal bone level mean average 0.22 +/- 0.87 mm (p < 0.05). There was only a slight mean difference compared to panoramic radiography but this was statistically insignificant, mean average 0.20 +/-1.35 mm (p > 0.05), and the periapical radiography had the lowest accuracy of radiographic methods, changed mean average -0.14 +/- 1.19 mm (p > 0.05). In summary, we can say that both bitewing and panoramic radiography are preferred to periapical images for crestal bone assessment. PMID- 19566028 TI - Intra-abdominal lymphangioma: an unusual lesion in an adult woman. PMID- 19566027 TI - Oral health status and treatment needs of a rural Indian fishing community. AB - A study was conducted to assess the oral health status and treatment needs of a rural Indian fishing community. The fishing community had poor access to dental care. The survey was carried out using the WHO 1997 criteria in natural daylight. Convenience sampling method was employed and all the subjects available were examined The results of this survey revealed that a large percentage of the population was afflicted with dental caries and periodontal disease. The unmet treatment need was found to be high in the study population. Appropriate oral health education and treatment is needed for this rural community. PMID- 19566029 TI - [The nature of the trophic tract in vascular plants]. AB - Modern data about the origin, structure and functions of plant plasmodesmata and "symplast" are reviewed. Etymology of "symplast" means the continuality of cell protoplasts in a plant body. Originally this term signified the continuality of sieve tubes only as a system of photosynthate transport and distribution. Content of their cavities seemed to be identical to cytoplasm. Later, the idea was shown to be a mistake. Sieve tube content is identical to vacuolar exudates. It has been shown that the plasmodesmata, vacuome of parenchyma and sieve tubes of phloem arised during prokaryote/eukaryote endosymbiogenesis as a transport network for photosynthate distribution. They continued to be the same during all the time of evolution of vascular plants. Functional role of the apoplast is similar when symplastic transport is blocked and apoplast becomes the compensatory channel. Both, the symplast and the apoplast, are continual in plant cell systems in contrast to cytoplasm whose continua-lity is secondary and has no serious significance as transport communication. It is not accorded with the phenomena of cell and tissue differentiation and cannot be accepted without additional evidences. The concept of plant symplast as a cytoplasm continuum could be reviewed under the influence of new facts. PMID- 19566030 TI - [Epigenetic nature of ageing]. AB - The idea proposed in this article is that a specific set of microRNAs expressing in stem cells can restore the initial profile of their epigenetic markers through RNAi-directed DNA methylation, and owing to that the pluripotent immortal status of these cells is supported unlimitedly and possibly minimum level of the mobile genomic elements activity is achieved. However, cell differentiation, starting with the earliest stages, must be accompanied with repression of genes of some microRNAs out of the primary set, otherwise these microRNAs would prevent expression of genes participating in the differentiation processes. Eventually, it results in that the cells slowly lose the repressive chromatin markers and this, sooner or later, will cause derepression of silent transposons and other mobile elements. This, accordingly, leads to the increase in DNA damage induced by these elements, and to following activation of cell systems of the DNA repair including mechanisms based on homologous recombination. In our opinion, these mechanisms cause not only DNA repair, but also unauthorized recombination on telomere capping structures, since they are pre-recombination structures. It is also possible that transposases in itself can initiate such recombination. As a result, the T-loops converse into rings and, accordingly, telomeres are shortened for the length of the lost circled DNA. This process can cause quick exhaustion of one or more cell telomeres and, therefore, subsequent senescence, cell cycle arrest and apoptosis of the cells, in which the illegitimate activation of recombination process becomes apparent. Apparently, large quantity of organism cells reaches with age the threshold of illegitimate activation of silent mobile genomic elements; subsequent apoptosis of most of these cells causes ageing as a biological phenomenon. PMID- 19566031 TI - [Effect of alpha-lipoic acid on the sensitivity of transformed fibroblasts to lysis by natural killer cells. Comparison with NAC action]. AB - The purpose of this study was to compare the effects of two antioxidants, alpha lipoic acid (ALA) and N-acetylcysteine (NAC) on the sensitivity of 3T3-SV40 fibroblasts to lytic activity of natural killer (NK) cells. ALA (1.25 mM) reduced significantly the fibroblast sensitivity in several hours, whereas NAC (10 mM) did not change it. Subsequent removal of the antioxidants from the cultivation medium resulted in gradual recovery of the sensitivity in the case of ALA and in complete loss of it in the case of NAC. Inactivation of gelatinase MMP-2 (matrix metalloproteinase) using pretreatment of the cells with the inhibitor of MMP, G6001, or specific antibodies to MMP-2 or MMP-9 resulted in decrease of 3T3-SV40 sensitivity to NK cells activity. This effect was similar to that of ALA, not to the NAC one. Pretreatment of NK cells with G6001 did not influence their lytic activity. The results obtained demonstrate that the direct antioxidant, NAC (having reduced thiol groups), and the indirect one, ALA (reducing thiol groups and acting as a direct antioxidant only inside the cell) activate principally different intracellular signal pathways. However, both NAC and ALA pathway includes inactivation of MMP-2. PMID- 19566032 TI - [Nongenomic mechanisms of progesterone]. AB - Rapid, independent of transcriptional effects of progesterone have been observed in various types of cells, tissues and species. In some biological systems, these nongenomic actions and associated with them signal transduction pathways are characterized in detail at molecular level. This review summarizes findings concerning the role of progestins in the regulation of such physiological functions and processes as meiotic maturation of fish and amphibian oocytes; growth and proliferation of normal and transformed cells of mammary gland; contraction of myometrium; survival and functional activity of granulose cells; sperm capacitation, acrosome reaction and hypermotility; immune function of T lymphocytes; survival and function of brain cells. The participation of several types of receptor proteins in the nongenomic progesterone signaling is discussed. They include the classic nuclear progesterone receptor, PR, the membrane progestin receptor, mPR, the progesterone membrane receptor component, PGMRC, the oxytocin receptor, OTR, and the GABA receptor, GABA(A). PMID- 19566033 TI - [Interrelation between the glycogen content in hepatocytes and their size in normal and cirrhotic rat liver]. AB - The dependence between the size of hepatocytes and glycogen content in them was investigated in normal and cirrhotic liver from fasting rats as well as from rats in 10 and 60 minutes after per os glucose administration. The cytophotometric method used allows glycogen and DNA contents determining in the same cell and its dry mass as well. It has been shown that the dry mass and the glycogen content in hepatocytes from normal and cirrhotic liver are proportional to genes dose. Normal liver hepatocytes of various ploidy classes show distinct correlation between the cell size and the glycogen content. No similar dependence is observed in hepatocytes population from the cirrhotic liver, apparently, because the liver lobule structure is disturbed, and heterogeneity of the hepatocytes microenvironment condition is increased. PMID- 19566034 TI - [Ah receptor-independent inhibition of gap junction intercellular communications in hepatoma cell culture 27 by polycyclic aromatic hydrocarbons]. AB - One of the systems that regulate tissue homeostatis is gap junction intercellular communications (GJIC). Inhibition of GJIC is widely used in experiments as a characteristic of tumor promotion. It is accepted that the down-regulation of GJIC is tightly related with the tumor-promoting properties of carcinogens. In this study, the effect of some carcinogenic polycyclic aromatic hydrocarbons on GJIC in cell cultures of hepatoma 27 lacking cytochrome P450 and Ah receptor was investigated. It was shown that inter 6 compounds studied only benzo/a/pyren and 3-methylcholanthrene were able to inhibit GJIC. We have concluded that an unknown factor is present in hepatoma cells and its interaction with some polycyclic aromatic hydrocarbons results in GJIC inhibition. The investigation of mutual effect of benzo/a/pyrene and non carcinogenic benzo/e/pyrene with similar structure has shown that GJIC inhibition by benzo/a/pyrene is at least double stepped. PMID- 19566035 TI - [The plate in the zone of oocyte and germinal epithelium contact in scyphomedusa Aurelia aurita binds antibodies to ZP-domain-containing protein mesoglein]. AB - Cnidaria are lower multicellular animals with the body consisting of two epithelial layers. An extracellular substance--mesoglea--is situated between epidermal and gastrodermal layers of these animals. Mesoglein is one of the major mesogleal proteins of adult medusa of Scyphozoan jellyfish Aurelia aurita. Search for the known domains in mesoglein amino acid sequence reveals prominent zona pellucida (ZP) domain (which was found at first in the mammal oocyte zona pellucida proteins), so the protein belongs to ZP family of extracellular matrix proteins and it is an early metazoan member of ZP-domain-containing protein family. However, nothing is known about oogenesis related ZP-domain proteins in the lower multicellular animals. Oogenesis in Scyphozoa is described poorly. In this work morphological features of the zone in contact area between the oocyte and the germinal epithelium were investigated in semi-fine sections: To make it more convenient we identified seven stages according to the oocyte size and the structure found in this area was named the plate. It was shown that the components of the plate bound specifically the antibodies against mesoglein. So it seems the plate material contains ZP-domain proteins. Electrophoresis and immunoblot results give evidence that the proteins immunologically related to mesoglein have a higher molecular mass. It might be due to either the posttranslational modifications of the precursors or that they represent other proteins of ZP-domain family in Cnidaria. PMID- 19566036 TI - [Murine embryonic stem cells as a model object for human embryonic stem cells research]. AB - During several last decades mouse embryonic stem cell (mESC) were used as a model object for the research of human embryonic stem cells (hESC). The validity of such approach is still not finally proved. At the same time there is a large amount of literary data available, which gives evidences of many substantial differences between these two cell types. The analysis of the literature has shown that there are differences concerning the regulation of proliferation, self renewal maintenance, and the regulation of differentiation of ESC. Thus, mESC can be considered as a model object for hESC studies only in some aspects of their biology. The alternative model objects, such as primate ESC, are also briefly discussed in this review. PMID- 19566037 TI - [Localization of satellite DNA and associated protein in respect to nucleolar precursor bodies in one- and two-cell mouse embryos]. AB - Nucleolar precursor bodies (NPB) are characteristic structures in the nuclei of one- and two cell mouse embryos. The alignment of centromeric (CEN) and pericentromeric (periCEN) chromosome regions to the chromatin layer surrounding NPB is known. Mus musculus 4 satellite DNA (satDNA) types are known to be located in CEN region--mouse minor satellite (MiSat) and mouse satellite 3 (MS3); and periCEN region--mouse major satellite (MaSat) and mouse satellite (MS4). We determined the localization of 4 types of mouse satDNA CEN and periCEN regions and associated proteins: RNA-helicase p68, SMC3, Rad21 subunits of the cohesin complex and SYCP3 subunit of the synaptonemal complex (SC). Partially flattened nuclei of the one- and two-cell embryos and embryos treated with ocadaic acids (OA) were used. Different satDNA fragments revealed distinct domains at the surface of NPB: periCEN MaSat was always localized in NPB more internally covering almost entire surface of NPB while CEN MiSat, MS3 and periCEN MS4 showed more peripheral localization. All 4 satDNA did not cover the entire areas of the NPB, indicating the presence of other DNA sequence involved in its formation. RNA helicase p68 and components of multiprotein cohesin and synaptonemal complexes are the necessary components of NPB. Our results support the opinion that NPB serve as a precursor of chromocenters. PMID- 19566038 TI - [Clinical and epidemiological study of lacrimal neoplasms]. AB - A cross-sectional mass epidemiological survey of 18,320 urban and rural citizens of the Republic of Tadjikistan revealed 31 patients with orbital tumors, of which lacrimal neoplasms were determined in 4 (0.022% in the general population and 12.9% of the identified orbital tumors). According to the referral data, the prevalence of lacrimal neoplasms was 1.03% of all the tumors of the organ of vision and 5.26% of the orbital neoplasms. Epithelial tumors were more common (52%), their malignant forms dominated over benign ones (55.7% versus 44.3%). Inflammatory processes (22.6%) as acute and chronic dacryoadenitis (9.5%) and pseudotumors (13.1) were the second most common conditions. Lymphoid tissue tumors (13.6%), cystoid masses (8.6%), and neoplasms of unclear genesis (1.4%) were noticeably less prevalent. The severest difficulties in recognizing lacrimal epithelial tumors were to clinically and morphologically identify what determined the specific features of the lacrimal structure and evident structural polymorphism. With the higher grade of anaplasia, the likelihood of a recurrence of a tumor and the rate of its growth increased (r = +0.83). The survival rates closely correlated with the higher rates of recurrent adenocarcinoma and adenocystic cancer (r = -0.87). PMID- 19566039 TI - [Immunogenetic criteria for uveitis development in the population of the Republic of Bashkortostan]. AB - Based on the association of certain histocompatibility Class I antigens with uveitis in patients from the Republic of Bashkortostan it has been proven that HLA B27 and B47 and haplotype A2B47 should be considered to be the markers responsible for the occurrence of uveitis in systemic and syndromic diseases. The greatest relative risk of herpetic anterior and posterior uveitis is associated with HLA B44 and B51 antigens; the risk of anterior and posterior uveitis is related to HLA B27 and HLA A28, A29 and B53, respectively. The findings permit improvement of the quality of early diagnosis of these diseases and are recommended for use in ophthalmological practice. PMID- 19566040 TI - [Investigation of scleral fibronectin in primary open-angle glaucoma: immunohistochemical study]. AB - The present investigation was first to study scleral fibronectin (FN) levels in primary open-angle glaucoma, by applying an immunohistochemical study. The enhanced accumulation of this glycoprotein was found in the deep and inner scleral layers. Active FN expression was ascertained to correspond to the scleral morphological changes characteristic for glaucoma (garneting of collagenous fibers, accumulation of free glycosamine glycans) and to be of focal nature. The findings show the role of FN in the mechanisms of scleral destruction in primary open-angle glaucoma. PMID- 19566041 TI - [Diagnosis and treatment of HLA-B27-associated uveitis]. AB - The purpose of the present investigation was to diagnose and to treat HLA-B27 antigen-associated uveitis in 102 patients (118 eyes) with anterior and posterior uveitis. HLA antigens were typed to specify the etiopathogenesis of uveitis. Along with this, the patients were examined by a rheumatologist, a rontgenologist, an immunologist, an urologist, a dermatovenereologist, a dentist, and an otorhinolaryngologist. The conventional ophthalmological study and eye B scanning were also performed. After complex therapy, there were improvements in the ocular state and overall somatic condition, as confirmed by the results of clinical studies. In 81 patients, the time of relapses reduced and the duration of remissions increased; persistent remission occurred in 15 patients. The course of the disease was unchanged in 7 cases. The pathogenetic role of B-27 antigen in the development of uveitis, as well as ankylosing spondylarthritis, seronegative spondylarthropathy, and other types of spondyloarthritis has not been explained so far. Notwithstanding this, the joint efforts of an oculist, a rheumatologist, and, not rarely, other specialists and the specific features of a clinical approach could provide a pronounced positive effect in treating the patients with HLA-27-associated uveitis. PMID- 19566042 TI - [Congenital glaucoma and thymomegaly: clinical and immunological characteristics]. AB - A comprehensive examination of children with thymomegaly and congenital glaucoma revealed systemic and local humoral immunological disorders mainly as immunodeficiency, a family history of the disease, susceptibility to infections and allergic diseases, the presence of malformations, severe glaucoma, and a complicated postoperative period, which are suggestive of the defective state of the immune system, which should be kept in mind while making a surgical treatment and using an immunomodulator therapy, by exercising a great caution. Such children require observations by a pediatrician, an oculist, and an immunologist and consultations given by other specialists. PMID- 19566043 TI - [Study of the accuracy of measure the critical flicker frequency]. AB - The possible accuracy of determining the critical flicker frequency (CFF) was assessed. According to the results of experimental studies, the individual accuracy of CFF determination was 0.7 to 1.3 Hz in a group of 30 examinees; the median distribution was 1.0 Hz; confidence interval for the median at 95% significance was 0.9 and 1.0 Hz. From the data of the experimental studies, it is recommended that a change step for a flicker frequency should be 0.5 Hz at mass CFF measurements. PMID- 19566044 TI - [Procedure for calculating the volume of the orbital fat to be removed at decompressive surgery in patients with endocrine ophthalmopathy]. AB - The paper gives data of computed tomography in 100 patients (148 orbits) with edematous exophthalmos (lipogenic and mixed types). There is a relationship of the higher degree of exophthalmos to the increased orbital soft tissue contents. A procedure is described for the individual calculation of the amount of removed orbital fat during compressive operations, which considers both the individual features of the anatomic structure and existing exophthalmos. PMID- 19566045 TI - [Ultrasound biomicroscopy in the monitoring of iridocorneal endothelial syndrome]. AB - Detailed analysis of scanning images obtained at ultrasound biomicroscopy (USBM) in 11 patients with iridocorneal endothelial syndrome revealed the USBM signs reflecting pathological changes in the anterior eyeball. The value of USBM is that the technique permits evaluation of the involvement of anatomic structures into the pathological process in corpore as this is the only mode of lifetime imaging of high-resolution silent eyeball areas. It is also important that USBM can differentiate the iridocorneal endothelial syndrome from other abnormalities with the same clinical picture. The authors recommend that USBM be used to diagnose and to monitor different types of the iridocorneal endothelial syndrome. PMID- 19566046 TI - [Comparative analysis of the results of microperimetry and conventional computed perimetry in health]. AB - Twenty-one healthy volunteers aged 21 to 37 years (21 eyes) were examined on a MP 1 microperimeter and a Humphrey HFA II-750i perimeter by the 10-2 program. The standard stimulation parameters used on the microperimeter revealed a wide plateau in the central visual field due to the limited stimulus intensity range of MP-1. The comparable data on both perimeters could be obtained only with small sized stimuli (Goldmann I). However, quantitative comparison demonstrated significant differences in the reduction of differential light thresholds from 2 degrees to 10 degrees from the fixation point. The studies have indicated that the technical characteristics of the MP-1 microperimeter does not allow one to elaborate algorithms for recounting the data to compare with those obtained on the Humphrey perimeter. It has been ascertained that the MP-1 microperimeter should not be used to solve the problems of conventional computed perimetry. PMID- 19566047 TI - [Minimal compression-load ophthalmoplethysmography in health and in primary open angle glaucoma]. AB - The investigation was undertaken to study intraocular blood flow in health and in primary open-angle glaucoma, by applying ophthalmoplethysmography with the minimum compression load on the eyeball. Sixty-nine patients (129 eyes) with primary open-angle glaucoma and 41 healthy individuals (82 eyes) were examined. In healthy individuals, the ophthalmoplethysmographic parameters of ocular volumetric blood flow averaged 7.3 mm3 for pulse volume and 493 mm3 for minute volume. There was a noticeable drop in volumetric ocular blood flow parameters in early primary open-angle glaucoma and their further decrease as a glaucomatous process progressed. The patients with primary open-glaucoma showed an inverse correlation of the volumetric hemodynamic parameters with the intraocular pressure. Worse temporary plethysmographic parameters were found in patients with normal intraocular pressure, which is an indirect indication of impaired choroidal blood microcirculation in glaucoma even within the normal range of intraocular pressure. PMID- 19566048 TI - [YAG laser reconstructive surgery of the anterior eye in children: results, indications, and optimal time]. AB - By analyzing the anatomic-reconstructive, optic, and functional results of 1526 YAG laser operations made by the author in children aged 2 months to 17 years who had congenital (65.99%), posttraumatic (23.53%), and postuveal (10.48%) abnormalities of the anterior eye, the author has developed indications and contraindications and determined the optimum time of reconstructive laser interventions in papillary membranes; anterior and posterior fusions; internal fistular atresia after trabeculectomy, in umbrella iris, deformity, dislocation, atresia of the pupil; in anterior chamber cysts, cataracts, and ectopic lens. The high efficiency (93.93%) and rather low traumatism show it expedient to extensively introduce laser reconstructive surgery into pediatric ophthalmological care as a worthy alternative to instrumental surgery, which yields a reconstructive effect without eye incision, in a large proportion (46.4%) of the children who have refused anesthesia essential for microsurgery. PMID- 19566050 TI - [Mycotic diseases of the lacrimal tract]. PMID- 19566049 TI - [Complex treatment for decompensated edematous exophthalmos]. AB - In endocrine ophthalmolopathy, there may be persistent dysfunction of the organ of vision due to corneal lesion or optical neuropathy. As an illustration of severe corneal lesion in edematous exophthalmos decompensation, the authors give their own observation that shows it necessary to use in such cases complex treatment comprising pulse therapy with glucocorticoids, followed by a long-term dosage reduction and switching over to its oral administration, as well radiotherapy and surgery. PMID- 19566051 TI - [Factors influencing ocular angiogenesis]. AB - The review of literature covers the topical aspects of the interaction of proangiogenic cytokines and angiogenic inhibitors, their implication in the pathogenesis of proliferative retinopathies occurring, among other diseases, in the presence of diabetes mellitus. The paper provides the historical facts of development of the angiogenic homeostasis theory that implies that there is a dynamic equilibrium between two antagonistic systems: proangiogenic and antiangiogenic. The major angiogenic inhibitors involved in the maintenance of the normal structure and function of the eye and in the processes linked with abnormal proliferation are enumerated. Particular emphasis is laid on the pigment epithelium-derived factor that is a potent antiangiogenic agent that is directly synthesized by ocular tissues and a vascular endothelial growth factor antagonist. The review of the literature considers the pathogenesis of diabetic retinopathy in the context of interaction of these two growth factors. PMID- 19566052 TI - [Current methods for treating the dry eye syndrome]. PMID- 19566053 TI - [Facial palsy: topographic and pathogenetic features of the lesion]. PMID- 19566054 TI - [Pathomorphological and histochemical manifestations of interaction of laser irradiation with tissues of a chorioretinal complex]. PMID- 19566055 TI - [Genetic resistance to malaria]. AB - Genetic resistance to malaria is associated with various genetic factors, including erythrocytic variability and variability of the genes involved into the pathogenetic process. Some genetic anomalies resulted from selective malaria pressure, which brought into existence different forms of hemoglobinopathies, glucose-6-phosphate dehydrogenase deficiency, and no Duffy antigens, and ovalocytosis, etc., which ensured varying malaria resistance. Cell adhesion is a major factor in the pathogenesis of malaria. Adhesion molecules express on the cellular membranes of the endothelium, platelets, macrophages, red blood cells and serve as binding receptors for membrane proteins PFRMP-1 of P. falciparum. Polymorphism of the CD36, ICAM-1, and PECAM1 genes can lower binding to blood vessel endothelial cells, which reduces the number of clinical forms of malaria. The high serum TNF-alpha level that is caused by mutation in the promoter of the TNF-alpha gene is associated with cerebral malaria. TNF-alpha enhances the endothelial expression of adhesion molecules, by increasing the adhesion of infected erythrocytes, including that in cerebral capillaries, by inducing in patients local thrombosis and inflammation with release of the cytokines--TNF alpha. The products of inflammatory infiltrates attack the endothelium, by leading to the imbibition of plasma and erythrocytes in brain tissue and causing a cerebral form of malaria. PMID- 19566056 TI - [Comparative characterization of the children's enteric protofauna in the Mid Urals in the early and late 20th century]. PMID- 19566057 TI - [The clinical and endoscopic state of the esophagus in patients with asthma concurrent with chronic opisthorchiasis]. AB - The frequency and pattern of manifestations of gastroesogageal reflux disease were studied in asthma concurrent with chronic opisthorchiasis. One hundred and fifty patients (52 males and 98 females) aged 14 to 60 years (mean age 43.18 +/- 10.6 years) who had atopic asthma of varying severity were examined. Group 1 comprised 74 asthmatic patients without opisthorchiasis; Group 2 included 76 asthmatic patients with opisthorchiasis; Group 3 consisted of 20 healthy individuals. According to the severity of asthma, all the patients from Group 1 and 2 were divided into 4 subgroups: mild, moderate, severe hormone-dependent and severe hormone-independent asthma. The patients with and without concurrent opisthorchiasis did not virtually differ in the severity pattern of asthma and its duration. Those with severe hormone-dependent asthma took systemic glucocorticosteroids in doses of not more than 15 mg/day. The scope of clinicoinstrumental and laboratory studies was determined by the presence of concurrent pathology: asthma and gastroduodenal diseases. Esophageal erosive lesions were encountered in a third of the asthmatic patients with opisthorchiasis and in every four patients in Group 1. In patients with severe asthma with and without opisthorchiasis, esophageal erosive lesions were revealed in 66.7%. The presence of concomitant Opisthorchis invasion in patients with asthma results in a significant increase in the clinical manifestations of gastroesophageal reflux disease and calls for the obligatory performance of therapy with antisecretory agents, prokinetics and necessitates dehelmintization during remitting asthma. PMID- 19566058 TI - [Effective dehelmintization therapy in patients with psoriasis concurrent with chronic opisthorchiasis, by using dietary supplements to vegetable food]. PMID- 19566059 TI - [Food fish parasitic contamination in the water reservoirs of the Novosibirsk Region]. PMID- 19566060 TI - [Parasitic diseases in the Kyrgyz Republic: epidemiological aspects]. AB - The paper analyzes the epidemiological situation associated with intestinal helminthic and protozoan diseases in the Kyrgyz Republic in the past 17 years (1991-2007). The analysis suggests that parasitic diseases are of wide spread in the republic. In the structure of infectious diseases, parasitic diseases account for 45%. Children less than 14 years of age are a risk group, whose proportion is 80%. Enteric nosological entities are prevalent in the structure of parasitic diseases. There has been a long-term trend for a rise in the incidence of echinococcosis and ascariasis and for a decrease in that of enterobiasis and hymenolepiasis. PMID- 19566061 TI - [Intestinal parasitic diseases in the Batken Region, Kyrgyz Republic]. AB - The paper compares the data of parasitological studies made according to the plan and epidemiological indications with the results of screening surveys of 1256 children from the Batken Region. The proportion of positive findings at the laboratories of therapeutic-and-prophylactic institutions was found to total 4.5%. According to the results of the screening surveys, the invasiveness was more than 100% with mixed invasions being kept in mind. Analysis of replies to questionnaires suggests that there was a high risk of infection in children. Thus, 84% of the respondents had bad habits; 62% did not wash fruits before eating. The Batken Region's population including 345984 persons was dehelmintized. The screening surveys carried out after dehelmintization revealed no ascarid eggs in 637 children. Health education on the prevention of parasitic diseases was simultaneously conducted. PMID- 19566062 TI - [Monitoring of arthropods of epidemiological and sanitary importance in the city of Voronezh]. AB - The paper analyzes the results of monitoring the number and species composition of arthropods, which are of epidemiological and sanitary important, in the city of Voronezh in 1998 to 2007. Many years' periodical observations have revealed the main places of breeding for blood sucking insects and ticks, their ecological features, including these under antropogenic stress. There is evidence for the changes in the species composition and the influence of some species on the recent epidemiological situation, which permits the approaches to and tactics of preventive measures to be corrected at the present budgetary stage oriented to outcome, with emphasis on the most significant sections of work. PMID- 19566063 TI - [Detection of arbovirus antigens in the mosquitoes and ticks inhabiting the Saratov Region]. AB - The paper gives the results of a study dealing with the detection of the antigens of arboviruses of West Nile, Sindbis, Batai, Crimean-Congo hemorrhagic fever, a serocomplex of Californian encephalitis in the field material gathered in the Saratov Region in 2000-2006. The bloodsucking arthropods inhabiting the region were shown to be actively involved in the circulation of arboviruses in natural biotopes. The conclusion that it is expedient to organize an annual monitoring of arbovirus-induced infections in the areas where positive findings have been notified is justified. PMID- 19566064 TI - [Metabolic features of continuous cell lines of mosquitoes]. AB - The metabolism of a continuous Ae. albopictus cell line (clone C6/36) was studied relative to the consumption of an environmental component by mammalian cells of the continuous lines Vero(B) and BNK-21 under steady-state cultivation conditions. The Ae. albopictus cells were found to utilize the amino acid glycine and on glycolysis they elaborated not more than (10...20)% of acid products (lactic acid) neutralized by sodium biocarbonate, suggesting that there were also other ways of using glucose. These features lead to the conclusion that the mosquito cells form an extracellular matrix. The findings were used to certify the culture of cells, to substantiate the composition of a cultivation medium, and to optimize the latter. PMID- 19566065 TI - [Role of burrow microbiocenosis in plague enzootia]. AB - The paper analyzes relationships of the plague bacilli to the representatives of different types of living organisms inhabiting the burrows. The authors give their own data on the qualitative and quantitative composition of indicoles of the burrow of little sousliks (Spermophillus pygmaeus). They assess the role of mutagenic agents in burrow microbiocenoses. PMID- 19566066 TI - [Experimental study of the inoculative transmission of Rickettsia typhi by gamasid mites (Gamasidae) Ornithonyssus bacoti]. AB - The authors' studies have established that the concentration of Rickettsia typhi may increase about 100-fold in the infected Ornithonyssus bacoti mites. At the time, when on feeding 20 to 200 adult mites on guinea-pigs and albino rats 4 to 36 days after inoculation, they did not transmit Rickettsia typhi on blood sucking. PMID- 19566067 TI - [Detection of Cryptosporidia oocysts in the excreta of agricultural animals and zoo predators]. PMID- 19566068 TI - [The clinical and epidemiological characteristics of vivax malaria in children]. PMID- 19566069 TI - [Hydatigerosis and strobilocercosis in the Republic of Belarus and their medical significance]. AB - The author presents the results of his own studies that were conducted during many years (1980-2007) and the data of Byelorussian helminthologists on the detection of the cestode of H. taeniaeformis and its larvae (strobilocercus), which induce hydatigerosis and strobilocercosis, respectively, in animals in the Republic of Belarus. This helminth (an imago and a larva) may parasite in man. Emphasis is laid on the fact that the residents of Belarus can be infected on chance ingestion of eggs of the cestode of H. taeniaeformis. PMID- 19566070 TI - [Peculiarities of some psycho-physiologic parameters change in pilots experiencing variable work and rest modes in long flights]. AB - The authors studied changes in some psycho-physiologic parameters in pilots during long flights with variable work and rest modes. Findings are peculiarities in changes of registered parameters--heart rate, attention, work load and performance in connection with variable work and rest modes in 7-hours piloting of aircraft. PMID- 19566071 TI - [Evaluating quality of medical examinations of oil and gas industry workers in medical institutions]. AB - The findings of independent reviews of the quality of periodic medical checkups of employees of oil and gas producing and refining facilities in medical institutions of various forms of ownership have been delivered. Actions to improve the quality of the periodic medical checkups have been proposed. PMID- 19566072 TI - [Occupational risk and health disorders criteria in metal mining industry workers]. AB - Evaluating occupational risk of health disorders in metal mining industry workers providing various ore extraction modes enabled to reveal early clinical, laboratory and functional markers of occupational and general diseases. PMID- 19566073 TI - [Social and psychologic factors of occupational stress forming in emotional exertion activities]. AB - The authors studied psycho-social factors influencing mental workers who bear high responsibilities for others' safety. Working in time deficit, inadequate worker's involvement into work process management, less career prospects, low possibility to improve work process, controversies in occupational tasks and roles were proved to be the most important occupational stress factors. Prevalence of occupational stress factors appeared not to be dependent on work intensity class. Findings were increasing influence of occupational motivation on physiologic parameters change in work process. PMID- 19566074 TI - [Structural and functional changes in heart under medical therapy in mental workers having arterial hypertension]. AB - Aim of the study was to study main structural and functional parameters of heart, evaluate efficiency of treatment (Egiloc, Cardilopin, their combination) in arterial hypertension patients. Examinations including ECG, EchoCG were performed initially, in 3 and 6 months after the treatment start. The study covered 126 patients with arterial hypertension II degree, who received the medium-dose therapy, and 34 apparently healthy individuals. PMID- 19566075 TI - [Scientific basis for complex anti-cancer program in area containing uranium extracting enterprises]. AB - The authors studied oncologic morbidity and mortality in area containing Navoisky mining metallurgy enterprise in Uzbekhistan. Evidence is that workers engaged into the main process in the enterprise and over at least 60% of working time subjected to occupational carcinogens have oncologic risk over 10 times exceeding that in the reference group. Workers engaged into the auxiliary processes and periodically subjected to occupational carcinogens over at most 40% of working time have relative oncologic risk ratio equal to 3. Oncologic morbidity in the workers having no direct exposure to occupational carcinogens is at most 2 times higher than that of general population. The authors specified and justified a goal-oriented complex anti-cancer program in the enterprise, for improving the activities of its oncologic service. PMID- 19566076 TI - [Influence of working environment factors on health state of road police officers controlling traffic (review of literature)]. AB - The analytic review demonstrated negative influence of polluted air on respiratory diseases formation, on development of pathologic changes in neuro endocrine and cardiovascular systems, kidneys and blood of traffic regulation officers. Special attention was put on possible carcinogenic and genotoxic influence of air polluted with chemicals in exhaust automobile gases. PMID- 19566077 TI - [Influence of nonindustrial risk factors on congenital abnormalities formation]. AB - Congenital abnormalities could result from exposure to occupational hazards. Epidemiologic study of nickel compounds influence on reproductive health in females engaged into nickel purification in Murmansk region enterprises did not reveal increased risk of the anomalies. The study was aimed to define influence of various risk factors connected not to work conditions, but to mother's health, bad habits, age, on congenital abnormalities in newborns. PMID- 19566078 TI - [Immunologic criteria of efficiency obtained from inclusion of high quality drinkable water "Rokadovskaya" into complex therapy of workers]. AB - The authors proved "Rokadovskaya" water to be efficient in complex therapy of workers with occupational and industrially mediated diseases. The effects included normalization and activation of antioxidant and phagocytic functions of peripheral neutrophils. Immunologic effects on immune cells and humoral immunity parameters were studied for various nosologic entities. Discussion covered mechanisms and prospects of immune correctors inclusion into complex therapy, according to contemporary concept of information role played by clusters in water structure. PMID- 19566079 TI - Systematical optimization of reverse-phase chromatography for shotgun proteomics. AB - We report the optimization of a common LC-MS/MS platform to maximize the number of proteins identified from a complex biological sample. The platform uses digested yeast lysate on a 75 microm internal diameter x 12 cm reverse-phase column that is combined with an LTQ-Orbitrap mass spectrometer. We first generated a yeast peptide mix that was quantified by multiple methods including the strategy of stable isotope labeling with amino acids in cell culture (SILAC). The peptide mix was analyzed on a highly reproducible, automated nanoLC-MS/MS system with systematic adjustment of loading amount, flow rate, elution gradient range and length. Interestingly, the column was found to be almost saturated by loading approximately 1 microg of the sample. Whereas the optimal flow rate ( approximately 0.2 microL/min) and elution buffer range (13-32% of acetonitrile) appeared to be independent of the loading amount, the best gradient length varied according to the amount of samples: 160 min for 1 microg of the peptide mix, but 40 min for 10 ng of the same sample. The effect of these parameters on elution peptide peak width is evaluated. After full optimization, 1012 proteins (clustered in 806 groups) with an estimated protein false discovery rate of approximately 3% were identified in 1 microg of yeast lysate in a single 160-min LC-MS/MS run. PMID- 19566080 TI - Chemical and enzymatic stability of amino acid prodrugs containing methoxy, ethoxy and propylene glycol linkers. AB - We evaluated the chemical and enzymatic stabilities of prodrugs containing methoxy, ethoxy and propylene glycol linkers in order to find a suitable linker for prodrugs of carboxylic acids with amino acids. l-Valine and l-phenylalanine prodrugs of model compounds (benzoic acid and phenyl acetic acid) containing methoxy, ethoxy and propylene glycol linkers were synthesized. The hydrolysis rate profile of each compound was studied at physiologically relevant pHs (1.2, 4, 6 and 7.4). Enzymatic hydrolysis of propylene glycol containing compounds was studied using Caco-2 homogenate as well as purified enzyme valacyclovirase. It was observed that the stability of the prodrugs increases with the linker length (propyl > ethyl > methyl). The model prodrugs were stable at acidic pH as compared to basic pH. It was observed that the prodrug with the aliphatic amino acid promoiety was more stable compared to its aromatic counterpart. The comparison between benzyl and the phenyl model compounds revealed that the amino acid side chain is significant in determining the stability of the prodrug whereas the benzyl or phenyl carboxylic acid had little or no effect on the stability. The enzymatic activation studies of propylene glycol linker prodrug in the presence of valacyclovirase and cell homogenate showed faster generation of the parent drug at pH 7.4. The half-life of prodrugs at pH 7.4 was more than 12 h, whereas in the presence of cell homogenate the half-lives were less than 1 h. Hydrolysis by Caco-2 homogenate generated the parent compound in two steps, where the prodrug was first converted to the intermediate, propylene glycol benzoate, which was then converted to the parent compound (benzoic acid). Enzymatic hydrolysis of propylene glycol containing prodrugs by valacyclovirase showed hydrolysis of the amino acid ester part to generate the propylene glycol ester of model compound (propylene glycol benzoate) as the major product. The amino acid prodrugs containing methoxy linker were the least stable while prodrugs containing propylene glycol linker were most stable. This work suggests that the propylene glycol linker is an optimal linker for amino acid prodrugs since it has good chemical stability and is enzymatically hydrolyzed to yield the parent drug. This approach can be further extended to other non-amino acid prodrugs and to provide a chemical handle to modify lead molecules containing carboxylic group(s). PMID- 19566081 TI - Universal DNA-based methods for assessing the diet of grazing livestock and wildlife from feces. AB - Because of the demand for controlling livestock diets, two methods that characterize the DNA of plants present in feces were developed. After DNA extraction from fecal samples, a short fragment of the chloroplastic trnL intron was amplified by PCR using a universal primer pair for plants. The first method generates a signature that is the electrophoretic migration pattern of the PCR product. The second method consists of sequencing several hundred DNA fragments from the PCR product through pyrosequencing. These methods were validated with a blind analysis of feces from concentrate- and pasture-fed lambs. The signature method allowed differentiation of the two diets and confirmed the presence of concentrate in one of them. The pyrosequencing method allowed the identification of up to 25 taxa in a diet. These methods are complementary to the chemical methods already used. They could be applied to the control of diets and the study of food preferences. PMID- 19566082 TI - Antifungal activity of CHE-23C, a dimeric sesquiterpene from Chloranthus henryi. AB - An antifungal compound was isolated from methanol extracts of stems and roots of Chloranthus henryi Hemsl. using ethyl acetate extraction and various chromatographic techniques. On the basis of spectroscopic analyses including mass and various NMR, the structure of the compound was identified as a dimeric sesquiterpene, CHE-23C. The compound showed potent antifungal activities (MICs = 1-32 microg/mL) in vitro against various phytopathogenic fungi such as Alternaria kikuchiana , Botrytis cinerea , Colletotrichum lagenarium , Magnaporthe grisea , Pythium ultimum , and Phytophthora infestans . In particular, it exhibited 91 and 100% disease-control activity in vivo against tomato late blight (P. infestans) and wheat leaf rust ( Puccinia recondita ) at concentrations of 33 and 100 microg/mL, respectively. The disease-control activity of this compound was stronger than that of the commercially available fungicide chlorothalonil, but weaker than that of dimethomorph. Therefore, the compound might serve as an interesting lead to develop effective antifungal agents. PMID- 19566083 TI - Antioxidant activity of wine pigments derived from anthocyanins: hydrogen transfer reactions to the dpph radical and inhibition of the heme-induced peroxidation of linoleic acid. AB - The consumption of red wine can provide substantial concentrations of antioxidant polyphenols, in particular grape anthocyanins (e.g., malvidin-3-O-beta-d glucoside (1)) and specific red wine pigments formed by reaction between anthocyanins and other wine components such as catechin (3), ethanol, and hydroxycinnamic acids. In this work, the antioxidant properties of red wine pigments (RWPs) are evaluated by the DPPH assay and by inhibition of the heme induced peroxidation of linoleic acid in acidic conditions (a model of antioxidant action in the gastric compartment). RWPs having a 1 and 3 moieties linked via a CH(3)-CH bridge appear more potent than the pigment with a direct 1 3 linkage. Pyranoanthocyanins derived from 1 reduce more DPPH radicals than 1 irrespective of the substitution of their additional aromatic ring. Pyranoanthocyanins are also efficient inhibitors of the heme-induced lipid peroxidation, although the highly hydrophilic pigment derived from pyruvic acid appears less active. PMID- 19566084 TI - Thermal effects on the stability and antioxidant activity of an acid polysaccharide conjugate derived from green tea. AB - A technique of high-performance gel permeation chromatography (HPGPC)-evaporative light-scattering detection and circular dichroism (CD) was developed for the measurement of thermal effects on the homogeneity and conformation of polymeric carbohydrate conjugates and was applied to an acid polysaccharide conjugate (GTa) isolated from the composite enzyme extract of green tea. Incubations in water at 40 and 70 degrees C for 1.0, 2.5, and 5.0 h have no effects on GTa. In contrast, when incubated in water for 1.0, 2.5, and 5.0 h at 98 degrees C, a single symmetrical peak corresponding to GTa in HPGPC was split into two adjacent peaks representing two different components formed, and CD spectra revealed an additional positive Cotton effect at 216 nm. To contribute toward our understanding of thermal effects of this polymeric carbohydrate conjugate on antioxidant activity, GTa and related heat-treated samples (GTa-HTI, GTa-HTII, and GTa-HTIII), the latter being obtained from 1.0, 2.5, and 5.0 h incubations at 98 degrees C, respectively, were subjected to the self-oxidation of 1,2,3 phentriol assay and found to have respective scavenging activities in a concentration-dependent manner. In comparison with GTa, the scavenging potency of heat-treated samples was similar at the dosage range of 50-300 microg/mL but became stronger with continually increasing concentration. Moreover, the present study also provides further insights into the optimal preparation of tea polysaccharide conjugates. PMID- 19566085 TI - Covalent insertion of antioxidant molecules on chitosan by a free radical grafting procedure. AB - In this work, the synthesis of gallic acid-chitosan and catechin-chitosan conjugates was carried out by adopting a free radical-induced grafting procedure. For this purpose, an ascorbic acid/hydrogen peroxide redox pair was employed as radical initiator. The formation of covalent bonds between antioxidant and biopolymer was verified by performing UV, FT-IR, and DSC analyses, whereas the antioxidant properties of chitosan conjugates were compared with that of a blank chitosan, treated in the same conditions but in the absence of antioxidant molecules. The good antioxidant activity shown by functionalized materials proved the efficiency of the reaction method. PMID- 19566086 TI - Digital analysis of protein properties by an ensemble of DNA quadruplexes. AB - Here we show how different principles developed in the area of molecular logic gates can be applied to diagnostic technologies for proteins. Simultaneous operation of YES NOT and PASS 1 logic gates, produced by a protein sensing ensemble of DNA G-quadruplexes, is used to encode concentration levels of medicinally important proteins. An AND logic gate is another example, where molecular computation can be used to follow the interaction between proteins and metal ions. Combination of molecular Boolean logic with combinatorial sensing is demonstrated as a general strategy to realizing small scale, real time diagnosis of a variety of protein samples. PMID- 19566087 TI - Chemical synthesis of air-stable manganese nanoparticles. AB - Elemental manganese has a complex crystal structure and unusual magnetic properties, making it an intriguing target for exploration in nanocrystalline form. However, because of its oxophilicity and the difficulty in reducing soluble metal salts to elemental Mn using the most common solution-phase reducing agents, it has been challenging to synthesize and stabilize elemental Mn nanoparticles using solution chemistry methods. Here we report the chemical synthesis of alpha Mn nanoparticles using n-butyllithium as a reducing agent. The nanoparticles have been characterized by powder XRD, TEM, electron diffraction, infrared spectroscopy (DRIFT), XPS, and SQUID magnetometry. An amorphous manganese oxide layer bound by oleate ligands helps to render the nanoparticles air-stable. The oxide-coated alpha-Mn nanoparticles are paramagnetic. PMID- 19566088 TI - Time-resolved study of surface spin effect on spin-lattice relaxation in Fe3O4 nanocrystals. AB - The role of surface spins in the relaxation of magnetization in optically excited Fe(3)O(4) nanocrystals has been investigated via time-resolved Faraday rotation measurements. The time scale of the magnetization recovery following the optically induced demagnetization increased from 250 to 350 ps as the size of the nanocrystals increased from 5 to 15 nm. From analysis of the data with a simple model relating the spin-lattice relaxation rate to the average spin-orbit coupling strength of the interior and surface spins, we estimated the relative efficiency of surface in spin-lattice relaxation with respect to the interior spins. Our analysis indicates that, in Fe(3)O(4) nanocrystals passivated with oleic acid, the surface is 3 times more efficient in spin-lattice relaxation than the interior. PMID- 19566089 TI - Folding DNA origami from a double-stranded source of scaffold. AB - Combined heat and chemical denaturation of double-stranded DNA scaffold strands in the presence of staple strands, followed by a sudden temperature drop and then stepwise dialysis to remove the chemical denaturant, leads to self-assembly of two distinct DNA-origami structures. PMID- 19566091 TI - Ni(0)-catalyzed formation of azaaluminacyclopentenes via azanickelacyclopentenes: a unique nickel/aluminum double transmetalation reaction. AB - Azaaluminacyclopentenes, which serve as useful precursors for gamma-substituted allylamines, were catalytically prepared from novel Ni-mediated three-component cyclocondensation of an alkyne, an imine, and AlMe(3). PMID- 19566090 TI - Fluorescent cassettes for monitoring three-component interactions in vitro and in living cells. AB - Two water-soluble "through-bond energy transfer cassettes" (TBET-cassettes) were prepared. They have good extinction coefficients at the donor part and transfer energy to the acceptor parts with good "overall quantum yields" (0.30 and 0.24 in pH 7.4 phosphate buffer). Fluorescence resonance energy transfer from one protein functionalized with an appropriate donor can be used to excite the TBET-cassettes on different proteins to probe protein-protein interactions under conditions that would not be possible for single-dye acceptor systems. PMID- 19566092 TI - Stabilization of iridium(IV) by monoanionic dialkyldiarylguanidinato ligands. AB - Electron-rich tris(guanidinato) complexes of Ir(III), [Ir{ArNC(NR(2))NAr}(3)] (where R = Me or Et; Ar = Ph or 4-MeC(6)H(4)), were synthesized from the respective [Ir{ArNC(NR(2))NAr}(C(8)H(14))(2)] precursors (C(8)H(14) = cis cyclooctene), are air-sensitive, and can be electrochemically oxidized in two one electron transfer steps. The first electron transfer is reversible and occurs at much lower potentials than typical for Ir(III). Chemical oxidation by [FeCp(2)]PF(6) afforded isolable, paramagnetic Ir(IV) compounds, [Ir{ArNC(NR(2))NAr}(3)]PF(6), which were characterized by analytical and spectroscopic methods and a single-crystal structure determination, demonstrating that Ir(IV) is accessible in a nitrogen-donor ligand environment. PMID- 19566094 TI - Making hydrogen from water using a homogeneous system without noble metals. AB - A photocatalytic noble metal-free system for the generation of hydrogen has been constructed using Eosin Y (1) as a photosensitizer, the complex [Co(dmgH)(2)pyCl](2+) (5, dmgH = dimethylglyoximate, py = pyridine) as a molecular catalyst, and triethanolamine (TEOA) as a sacrificial reducing agent. The system produces H(2) with an initial rate of approximately 100 turnovers per hour upon irradiation with visible light (lambda > 450 nm). Addition of free dmgH(2) greatly increases the durability of the system addition of 12 equiv of dmgH(2) (vs cobalt) to the system produces approximately 900 turnovers of H(2) after 14 h of irradiation. The rate of H(2) evolution is maximum at pH = 7 and decreases sharply at more acidic or basic pH. Spectroscopic study of photolysis solutions suggests that hydrogen production occurs through protonation of a Co(I) species to give a Co(III) hydride, which then reacts further by reduction and protolysis to give Co(II) and molecular hydrogen. PMID- 19566093 TI - ENDOR spectroscopy shows that guanine N1 binds to [4Fe-4S] cluster II of the S adenosylmethionine-dependent enzyme MoaA: mechanistic implications. AB - The S-adenosylmethionine-dependent enzyme MoaA, in concert with MoaC, catalyzes the first step of molybdenum cofactor biosynthesis, the conversion of guanosine 5'-triphosphate (5'-GTP) into precursor Z. A published X-ray crystal structure of MoaA with the substrate 5'-GTP revealed that the substrate might be bound to the unique iron of one of two 4Fe-4S clusters through either or both the amino and N1 nitrogen nuclei. Use of 35 GHz continuous-wave ENDOR spectroscopy of MoaA with unlabeled and (15)N-labeled substrate and a reduced [4Fe-4S](+) cluster now demonstrates that only one nitrogen nucleus is bound to the cluster. Experiments with the substrate analogue inosine 5'-triphosphate further demonstrate that it is the N1 nitrogen that binds. Two of the more distant nitrogen nuclei have also been detected by 35 GHz pulsed ENDOR spectroscopy, allowing a rough approximation of their distances from the cluster to be calculated. Combining this information with the crystal structure, we propose that the guanine base adopts the enol tautomer as N1 binds to Fe4 and the O6-H hydroxyl group forms a hydrogen bond with S4 of the 4Fe-4S cluster, and that this binding-induced tautomerization may have important mechanistic ramifications. PMID- 19566095 TI - High-throughput screening of nanoparticle catalysts made by flame spray pyrolysis as hydrocarbon/NO oxidation catalysts. AB - We describe here the use of liquid-feed flame spray pyrolysis (LF-FSP) to produce high surface area, nonporous, mixed-metal oxide nanopowders that were subsequently subjected to high-throughput screening to assess a set of materials for deNO(x) catalysis and hydrocarbon combustion. We were able to easily screen some 40 LF-FSP produced materials. LF-FSP produces nanopowders that very often consist of kinetic rather than thermodynamic phases. Such materials are difficult to access or are completely inaccessible via traditional catalyst preparation methods. Indeed, our studies identified a set of Ce(1-x)Zr(x)O(2) and Al(2)O(3) Ce(1-x)Zr(x)O(2) nanopowders that offer surprisingly good activities for both NO(x) reduction and propane/propene oxidation both in high-throughput screening and in continuous flow catalytic studies. All of these catalysts offer activities comparable to traditional Pt/Al(2)O(3) catalysts but without Pt. Thus, although Pt-free, they are quite active for several extremely important emission control reactions, especially considering that these are only first generation materials. Indeed, efforts to dope the active catalysts with Pt actually led to lower catalytic activities. Thus the potential exists to completely change the materials used in emission control devices, especially for high-temperature reactions as these materials have already been exposed to 1500 degrees C; however, much research must be done before this potential is verified. PMID- 19566096 TI - One-step synthesis of core-shell (Ce0.7Zr0.3O2)(x)(Al2O3)(1-x) [(Ce0.7Zr0.3O2)@Al2O3] nanopowders via liquid-feed flame spray pyrolysis (LF FSP). AB - We report here the synthesis of Ce(x)Zr(1-x)O(2) and (Ce(0.7)Zr(0.3)O(2))(x)(Al(2)O(3))(1-x) core-shell nanopowders in a single step by liquid-feed flame spray pyrolysis (LF-FSP) of the metalloorganic precursors, Ce(O(2)CCH(2)CH(3))(3)(OH), alumatrane [N(CH(2)CH(2)O)(3)Al], and Zr(O(2)CCH(2)CH(3))(2)(OH)(2). Solutions of all three precursors in ethanol with ceramic yields of 2.5 wt% were aerosolized with O(2), combusted at temperatures above 1500 degrees C, and rapidly quenched at approximately 1000 degrees C/ms to form Ce(x)Zr(1-x)O(2) and (Ce(0.7)Zr(0.3)O(2))(x)(Al(2)O(3))(1-x) nanopowders of selected compositions, at rates of 50-100 g/h. The resulting, as-processed, materials are unaggregated nanopowders with average particle sizes (APSs) < 20 nm and corresponding specific surface areas of 30-50 m(2)/g. The as-processed powders were characterized in terms of phase, particle size, specific surface area, compositions, and morphology by XRD, BET, DLS, SEM, TEM, XPS, TGA-DTA, and FT-IR. LF-FSP provides access to binary Ce(x)Zr(1-x)O(2) nanopowders and ternary (Ce(0.7)Zr(0.3)O(2))(x)(Al(2)O(3))(1-x) nanopowders in one step. The obtained Ce(0.7)Zr(0.3)O(2) powders are solid solutions with a cubic phase. In contrast, LF-FSP of mixtures of the three precursors at specific compositions [x = 0.5, 0.7 for (Ce(0.7)Zr(0.3)O(2))(x)@(Al(2)O(3))(1-x)] provide core-shell nanopowders in a single step. The most reasonable explanation is that there are differences in the rates of condensation, nucleation and miscibility between the gas phase ions that form the Ce(x)Zr(1-x)O(2) solid solutions and those that condense to delta Al(2)O(3) during processing. These as-produced materials are without microporosity at surface areas of > or = 30 m(2)/g. Evidence is presented suggesting the formation of (Ce/Zr)(3+) species in the as-processed (Ce(0.7)Zr(0.3)O(2))(x)(Al(2)O(3))(1-x) core-shell materials. An accompanying paper indicates that these materials offer significant and novel catalytic activities for hydrocarbon oxidation and deNO(x) processes without using platinum as a co-catalyst. PMID- 19566098 TI - Stabilization of three-way junctions of DNA under molecular crowding conditions. AB - We examined the effects of molecular crowding conditions on the structures and thermodynamics of three-way junctions (TWJs) of DNA. Structural analysis utilizing gel electrophoresis and circular dichroism spectroscopy showed that the designed DNAs folded into TWJ structures in the presence of Na(+) and Mg(2+) under both dilute and molecular crowding conditions with polyethylene glycol 200 (PEG 200). From the thermodynamic parameters evaluated by UV melting techniques in the absence and presence of 5 mM Mg(2+) under dilute and molecular crowding conditions, it was clear that Mg(2+) stabilized all TWJs under the dilute condition, although the extent of stabilization depended on the stacking partners of TWJs. For example, thermodynamic stability (-DeltaG(o) (37)) of A/B-stacked TWJs (A, B, and C are the three helices of TWJ, and among these helices, A and B are stacked together) increased from 3.7 to 5.6 kcal/mol by the addition of 5 mM Mg(2+), while that of A/C-stacked TWJs (A and C are stacked together) increased only from 3.0 to 3.7 kcal/mol. Molecular crowding with PEG 200 destabilized the whole TWJ consisting of a junction point and three helical duplex arms. Crowding agents such as PEG 200 can affect the stability of DNA by modulating its hydration. To explore the crowding effects on the junction point, we evaluated the number of water molecules associated with the whole TWJ as well as the individual arms, and we found that the number of water molecules taken up by the whole TWJ was significantly smaller than the sum of the individual arms. These results show the dehydration from the junction point of the TWJ structure. Therefore, molecular crowding should be favorable for the junction point of TWJ structure and unfavorable for the duplex structure. To prove this concept, we designed truncated TWJ structures that folded into a bimolecular duplex under the dilute condition. With increasing concentrations of PEG 200 from 0 to 30 wt %, the fraction of truncated TWJ structures gradually increased, and that of the bimolecular duplex structure decreased, even in the absence of Mg(2+). We concluded that a cell-mimicking condition, in which the activity of water decreases and hydration becomes less favorable, might facilitate the formation of junction structures in comparison with duplexes. PMID- 19566097 TI - Rapid cross-linking of an RNA internal loop by the anticancer drug cisplatin. AB - Cisplatin is the most prominent member of a series of platinum(II) antitumor drugs that demonstrate activity based on binding to adjacent purines on genomic DNA. The interactions between cisplatin and alternate biomolecules, including chemically similar RNA, are less understood than are those for DNA. In order to investigate potential implications of platinum(II) drug binding to a structurally complex RNA, we have characterized the reaction between cisplatin and the internal loop of a 41-nucleotide subdomain derived from the U2:U6 spliceosomal RNAs. This "BBD" RNA subdomain consists of a hairpin structure containing a purine-rich asymmetric internal loop. Aquated cisplatin is found to cross-link G nucleobases on opposing sides of the internal loop, forming an intramolecular internal loop cross-link in BBD and an analogous intermolecular cross-link in a two-piece construct containing the same internal loop sequence. The two opposing guanine residues involved in the cross-link were identified via limited alkaline hydrolysis. The kinetics of aquated cisplatin binding to the BBD RNA, a related RNA hairpin, and its DNA hairpin analogue were investigated in an ionic background of 0.1 M NaNO(3) and 1 mM Mg(NO(3))(2). Both BBD and the RNA hairpin react 5-6-fold faster than the DNA hairpin, with calculated second-order rate constants of 2.0(2), 1.7(3), and 0.33(3) M(-1) s(-1), respectively, at 37 degrees C, pH 7.8. MALDI-MS data corroborate the biochemical studies and support a model in which kinetically preferred platinum binding sites compete with less reactive sites in these oligonucleotides. Taken together, these data indicate that cisplatin treatment has potential to create internal loop and other unusual cross links in structurally complex RNAs, on a time scale that is relevant for RNA dependent biological processes. PMID- 19566099 TI - Temperature dependence of "elementary processes" in doping semiconductor nanocrystals. AB - Controlled doping is a critical step toward various unique nanostructures. This report shall demonstrate that doping chemistry of colloidal nanocrystals is much more complex than what has been proposed in the existing experimental and theoretical reports. Four individual processes, namely "surface adsorption", "lattice incorporation", "lattice diffusion", and "lattice ejection", will be identified, each of which possesses its own critical temperature. A given type of host nanocrystals can be switched from being impossible to dope to becoming successfully doped. The key is to program the reaction temperature to accommodate all elementary processes. PMID- 19566100 TI - DNA repair and DNA triplet repeat expansion: the impact of abasic lesions on triplet repeat DNA energetics. AB - Enhanced levels of DNA triplet expansion are observed when base excision repair (BER) of oxidative DNA base damage (e.g., 8-oxo-dG) occurs at or near CAG repeat sequences. This observation suggests an interplay between processing mechanisms required for DNA repair and expansion pathways that yield genotypes associated with many neurological/developmental disorders. It has been proposed that DNA expansion involves the transient formation within the triplet repeat domains of non-native slipped DNA structures that are incorrectly processed by the BER machinery of repair during DNA synthesis. We show here that replacement within a triplet repeat bulge loop domain of a guanosine residue by an abasic site, the universal BER intermediate, increases the population of slipped/looped DNA structures relative to the corresponding lesion-free construct. Such abasic lesion-induced energetic enhancement of slipped/looped structures provides a linkage between BER and DNA expansion. We discuss how the BER machinery of repair may be influenced by abasic-induced energetic alterations in the properties of regions proximal to and/or within triplet repeat domains, thereby potentially modulating levels of DNA expansion. PMID- 19566101 TI - Universal translators for nucleic acid diagnosis. AB - Defined broadly, molecular translators are constructs that can convert any designated molecular input into a unique output molecule. In particular, the development of universal nucleic acid translators would be of significant practical value in view of the expanding biomedical importance of gene diagnostics. Currently, diagnostic assays for nucleic acids must be individually developed and optimized for each new sequence because inputs for one assay are sequence-specific and are therefore incompatible with any other assay designed for the detection of a different nucleic acid. However, if a desired nucleic acid sequence could be translated in vitro into a predetermined nucleic acid output for which there is already a known diagnostic assay, then that single assay could be easily adapted to detect nearly any strand. Here we investigate PCR independent isothermal molecular translation strategies that function without the need for post-translation purification and can be implemented with commercially available components. Translation yields up to 96% are obtained in 5 min at room temperature with minimal background reaction (<1%) and with discrimination of single nucleotide polymorphisms in the input sequence. Furthermore, we apply these translators to adapt a high-gain HIV diagnostic system for high-throughput detection of hepatitis C, avian influenza (H5N1), and smallpox without making changes to the underlying assay. Finally, we show the feasibility of translating small-molecule interactions into nucleic acid outputs by demonstrating the utility of a DNA aptamer for translating adenosine into a readily detectable output DNA sequence. Additionally, equilibrium expressions are described in order to facilitate rational engineering of aptameric translators for label-free detection of any molecule that an aptamer can recognize. PMID- 19566102 TI - trans-2 addition pattern to power charge transfer in dendronized metalloporphyrin C60 conjugates. AB - Coordinating different transition metals--manganese(III), iron(III), nickel(II), and copper(II)--by a dendronized porphyrin afforded a new family of redox-active metalloporphyrins to which C(60) was attached as a ground-state electron acceptor. Such a strategy introduced an additional center of redoxactivity, that is, a change of the oxidation state of the metal. Cyclic voltammetry and absorption/fluorescence measurements provided support for mutual interactions between the redox-active constituents in the ground state. In particular, slightly anodic shifted reduction potentials/cathodic shifted oxidation potentials and the occurrence of new charge transfer features in the 700-900 nm range prompt to sizable electronic coupling in the range of 300 cm(-1). Photophysical means--steady-state/time-resolved fluorescence and transient absorption measurements--shed light on the excited-state interactions. To this end, we have added pulse radiolytic investigations to characterize the radical cation (i.e., metalloporphyrins) and radical anion (i.e., fullerene) characteristics. Pi-pi stacking of the excited state electron donor and the electron acceptor is key to overcome the intrinsically fast deactivation of the excited states in these metalloporphyrins and to power an exothermic charge transfer. The lifetimes of the rapidly and efficiently generated radical ion pair states, which range from 15 to >3000 ps, revealed several important trends. First, they were found to depend on the solvent polarity. Second, the nature of the transition metal plays a similarly decisive role. It is important that the product of charge recombination, namely tripmultiplet excited states versus ground state, had a great impact. Finally, a correlation between the charge transfer rate (i.e., charge separation and charge recombination) and the free energy change for the underlying reaction reveals a parabolic dependence with parameters of the reorganization energy (0.84 eV) and electronic coupling (70 cm( 1)) closely resembling that seen for the zinc(II) and free base analogues. PMID- 19566103 TI - One-pot synthesis of platinum-based nanoparticles incorporated into mesoporous niobium oxide-carbon composites for fuel cell electrodes. AB - Catalyst-electrode design is crucial for the commercialization and widespread use of polymer electrolyte membrane fuel cells. There are considerable challenges in making less expensive, more durable, and more active catalysts. Herein, we report the one-pot synthesis of Pt and Pt-Pb nanoparticles incorporated into the pores of mesoporous niobium oxide-carbon composites. The self-assembly of block copolymers with niobium oxide and metal precursors results in an ordered mesostructured hybrid. Appropriate heat treatment of this hybrid produces highly crystalline, well-ordered mesoporous niobium oxide-carbon composites with Pt (or Pt-Pb) nanoparticles incorporated into the mesopores. The in situ-generated graphitic-like carbon material prevents the collapse of the mesostructure, while the metal oxide crystallizes at high temperatures and enhances the electrical conductivity of the final material. Formic acid electrooxidation with this novel material shows 4 times higher mass activities (3.3 mA/microg) and somewhat lower onset potentials (-0.24 V vs Ag/AgCl) than the best previously reported values employing Pt-Pb intermetallic nanoparticles supported on conducting carbon (0.85 mA/microg and -0.18 V, respectively). PMID- 19566106 TI - Multidisciplinarity goes mainstream: Orlando meeting of the Pediatric Pharmacy Advocacy Group focuses attention on the need for multidisciplinary networks. PMID- 19566107 TI - Non-infectious pediatric uveitis: an update on immunomodulatory management. AB - Pediatric non-infectious uveitis remains a rare but potentially sight-threatening group of diseases. However, early screening and treatment can improve outcomes. No single agent has proven to be efficacious in all cases. A wide variety of long term immunomodulatory treatments are available; these agents differ in both their potency and side effect profiles. Corticosteroids remain an extremely valuable form of treatment in the short-term management of uveitis. Other major groups of immunomodulatory treatments include the calcineurin inhibitors and antimetabolites such as methotrexate, which is frequently used as the first-line agent. The biologics, including anti-tumor necrosis factor agents and interferons, are newer and potentially very useful therapies although side effects limit their use. Successful outcomes may be achieved with appropriate immunosuppressant therapy given early in the disease, although clinical trials are required to define the true efficacy of this strategy. PMID- 19566108 TI - Variations in amoxicillin pharmacokinetic/pharmacodynamic parameters may explain treatment failures in acute otitis media. AB - Pharmacokinetic/pharmacodynamic (PK/PD) modeling and Monte Carlo simulations suggest that amoxicillin should rarely fail as therapy for Streptococcus pneumoniae and Haemophilus influenzae acute otitis media (AOM) infections except when the S. pneumoniae are highly penicillin resistant or the H. influenzae are beta-lactamase producing. However, important and not infrequent exceptions to this expectation have been described. The objective of this review was to define the biologic variations in amoxicillin PK/PD parameters for the treatment of AOM in children and assess whether these variations could explain why the commonly employed amoxicillin PK/PD model is imperfect in predicting outcome for every patient in this clinical setting. To this end, a literature search of MEDLINE (1966-2006) and EMBASE (1974-2006) was conducted to identify studies that evaluated ampicillin or amoxicillin intestinal absorption, serum concentrations, and/or middle ear fluid (MEF) concentrations. Analysis of studies identified for review showed that the intestinal bioavailability of amoxicillin depends on passive diffusion and a saturable 'pump' mechanism that produces variable serum concentrations of the antibacterial agent. Indeed, substantial differences from patient to patient in serum (5- to 30-fold) and MEF (up to 20-fold) concentrations of amoxicillin occur following oral administration, and 15-35% of children have no detectable amoxicillin in MEF. These findings suggest that variability in PK/PD parameters may impact amoxicillin concentrations in serum and MEF, possibly explaining some AOM treatment failures. PMID- 19566109 TI - Potential vitamin-drug interactions in children: at a pediatric emergency department. AB - BACKGROUND: A significant increase in vitamin use has been observed in recent years and interactions between vitamins and medications have been reported. OBJECTIVE: To determine the frequency and types of potential interactions between vitamins and medications in children arriving at a large tertiary, pediatric emergency department. We also compared family characteristics of children with potential interactions with those of children with no potential interactions, in order to determine children at a higher risk. METHODS: A cross-sectional study in which a survey was conducted of parents/caregivers and/or patients aged 0-18 years registered at a large pediatric emergency department in Canada. A total of 1804 families underwent a face-to-face interview. The main outcome measure was the rate of potential vitamin interactions in the preceding 3 months. RESULTS: A considerable number of patients (11% of our cohort) had potential vitamin medication interactions in the preceding 3 months, which could theoretically result in adverse events, and over one-third of these children had more than one potential interaction. Patients with potential interactions and their parents were significantly older (p < 0.001 for the child and mother, p = 0.02 for the father), the children were much more likely to have a chronic illness (p < 0.001) and concurrently receive prescribed or over-the-counter medication (p < 0.001), and more children with potential interactions were completely immunized (p = 0.02). The child's sex, parental education, employment status, family income, and primary language spoken at home were not associated with potential interactions. CONCLUSIONS: Taking into account the high rate of potential vitamin-drug interactions, especially among older children and patients with chronic illness, parents and healthcare providers need to balance the potential benefit of concurrent vitamin-medication use with its potential harms. PMID- 19566110 TI - Caspofungin: in pediatric patients with fungal infections. AB - Caspofungin is the first echinocandin to be approved for the treatment of fungal infections in pediatric patients. The antifungal properties of caspofungin result from interference with fungal cell-wall integrity. In vitro, caspofungin is fungicidal against Candida spp. and fungistatic against Aspergillus spp., but has little or no fungicidal or fungistatic activity against Cryptococcus neoformans, the Zygomycetes, Fusarium spp., or Trichosporon beigelii. Caspofungin was effective as empirical antifungal therapy in pediatric patients with persistent fever and neutropenia. Almost half (46%) of caspofungin recipients and one-third (32%) of liposomal amphotericin B recipients achieved an overall favorable response in a randomized, double-blind trial. Caspofungin was also effective in pediatric patients with fungal infections (invasive candidiasis, invasive aspergillosis refractory to or intolerant of standard antifungal agents, or esophageal candidiasis). Positive responses to treatment were seen in 30 of 37 patients with invasive candidiasis, 5 of 10 patients with invasive aspergillosis, and in the one patient with esophageal candidiasis, in a noncomparative, open label trial. Caspofungin was generally well tolerated in the clinical trials in pediatric patients with febrile neutropenia requiring empirical antifungal treatment, or with fungal infections. Few caspofungin recipients reported serious drug-related adverse events or discontinued treatment as a result of drug-related adverse events. PMID- 19566111 TI - Micafungin: a review of its use in the prophylaxis and treatment of invasive Candida infections in pediatric patients. AB - Intravenous micafungin (Mycamine; Funguard) is an echinocandin indicated in Japan and the EU for the treatment of pediatric patients (including neonates) with invasive candidiasis and as prophylaxis against Candida infection in pediatric patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT). In the EU, micafungin is also indicated in pediatric patients who are expected to have neutropenia for >/=10 days. In Japan, children may also receive micafungin for the treatment of, or as prophylaxis against, invasive Aspergillus infection. Micafungin is not currently approved for use in pediatric patients in the US. Micafungin has very good antifungal activity against a wide range of Candida spp. in vitro. It has a favorable pharmacokinetic profile allowing for once-daily administration, has few drug-drug interactions, and reports of resistance are rare. The results of pediatric substudies indicate that intravenous micafungin is effective in a majority of patients for the treatment of candidemia and other types of invasive candidiasis, and provides effective prophylaxis against invasive fungal infections in pediatric patients undergoing HSCT. The tolerability profile of micafungin in pediatric patients was generally acceptable. In the EU, micafungin is indicated for use when other antifungal medications are not appropriate. Therefore, micafungin provides an alternative to other antifungal agents used in the management of candidemia and invasive candidiasis in pediatric patients, or as prophylaxis against fungal infections in pediatric patients undergoing HSCT. PMID- 19566112 TI - Clinical pharmacokinetics and pharmacodynamics of solifenacin. AB - The succinate salt of solifenacin, a tertiary amine with anticholinergic properties, is used for symptomatic treatment of overactive bladder. Solifenacin peak plasma concentrations of 24.0 and 40.6 ng/mL are reached 3-8 hours after long-term oral administration of a 5 or 10 mg solifenacin dose, respectively. Studies in healthy adults have shown that the drug has high absolute bioavailability of about 90%, which does not decrease with concomitant food intake. Solifenacin has an apparent volume of distribution of 600 L, is 93-96% plasma protein bound, and probably crosses the blood-brain barrier. Solifenacin is eliminated mainly through hepatic metabolism via cytochrome P450 (CYP) 3A4, with about only 7% (3-13%) of the dose being excreted unchanged in the urine. Solifenacin metabolites are unlikely to contribute to clinical solifenacin effects. In healthy adults, total clearance of solifenacin amounts to 7-14 L/h. The terminal elimination half-life ranges from 33 to 85 hours, permitting once daily administration. Urinary excretion plays a minor role in the elimination of solifenacin, resulting in renal clearance of 0.67-1.51 L/h. Solifenacin does not influence the activity of CYP1A1/2, 2C9, 2D6 and 3A4, and shows a weak inhibitory potential for CYP2C19 and P-glycoprotein in vitro; however, clinical drug-drug interactions with CYP2C19 and P-glycoprotein substrates are very unlikely. Exposure to solifenacin is increased about 1.2-fold in elderly subjects and about 2-fold in subjects with moderate hepatic and severe renal impairment, as well as by coadministration of the potent CYP3A4 inhibitor ketoconazole 200 mg/day. The full therapeutic effects of solifenacin occur after 2-4 weeks of treatment and are maintained upon long-term therapy. Although solifenacin pharmacokinetics display linearity at doses of 5-40 mg, no obvious dose dependency was observed in efficacy and tolerability studies. The efficacy of solifenacin (5 or 10 mg/day) is at least equal to that of extended-release (ER) tolterodine (4 mg/day) in reducing the mean number of micturitions per 24 hours and urgency episodes, and in increasing the volume voided per micturition. Solifenacin (5 mg/day) appears to be superior to ER tolterodine (4 mg/day) in reducing incontinence episodes (mean -1.30 vs -0.90, p = 0.018) and is superior to propiverine (20 mg/day) at the dose of 10 mg/day in reducing urgency (-2.30 vs -2.78, p = 0.012) and nocturia episodes. Based on withdrawal rates due to adverse effects during the 52 week treatment period, solifenacin appears to have better tolerability than immediate-release (IR) oxybutynin 10-15 mg/day and IR tolterodine 4 mg/day. With regard to the pharmacokinetics of solifenacin, and for safety reasons, doses exceeding 5 mg/day are not recommended for patients with moderate hepatic impairment (Child-Pugh score 7-9), patients with severe renal impairment (creatinine clearance <30 mL/min) and subjects undergoing concomitant therapy with CYP3A4 inhibitors. PMID- 19566113 TI - Clinical pharmacokinetic and pharmacodynamic profile of cinacalcet hydrochloride. AB - Cinacalcet hydrochloride (cinacalcet) is a calcimimetic approved for the treatment of secondary hyperparathyroidism in patients with chronic kidney disease (CKD) receiving dialysis and for the treatment of hypercalcaemia in patients with parathyroid carcinoma. Following oral administration, peak plasma concentrations of cinacalcet occur within 2-6 hours. The absolute bioavailability is 20-25%, and administration of cinacalcet with low- or high-fat meals increases exposure (area under the plasma concentration-time curve from time zero to infinity [AUC(infinity)]) 1.5- to 1.8-fold. Cinacalcet has no significant interaction with calcium carbonate or sevelamer hydrochloride, phosphate binders commonly used in the treatment of patients with CKD receiving dialysis. The terminal elimination half-life is 30-40 hours, and steady-state concentrations are achieved within 7 days. The pharmacokinetics of cinacalcet are dose proportional over the dose range of 30-180 mg. The pharmacokinetic profile of cinacalcet is not notably affected by varying degrees of renal impairment. The pharmacokinetics of cinacalcet are comparable between healthy subjects, patients with primary hyperparathyroidism and patients with secondary hyperparathyroidism with reduced renal function (including those patients with secondary hyperparathyroidism receiving dialysis). Additionally, the pharmacokinetics of cinacalcet are similar in patients with secondary hyperparathyroidism receiving haemodialysis and patients with secondary hyperparathyroidism receiving peritoneal dialysis. Mild hepatic impairment does not affect the pharmacokinetics of cinacalcet, whereas moderate or severe hepatic impairment increases the exposure (AUC(infinity)) by approximately 2- and 4-fold, respectively. Age, sex, bodyweight and race do not notably affect the pharmacokinetics of cinacalcet. Cinacalcet is extensively metabolized by multiple hepatic cytochrome P450 (CYP) enzymes (primarily 3A4, 2D6 and 1A2) with <1% of the parent drug excreted in the urine. Dose adjustments of cinacalcet may be necessary, and parathyroid hormone (PTH) and serum calcium concentrations should be closely monitored if a patient initiates or discontinues therapy with a strong CYP3A4 inhibitor (e.g. ketoconazole, erythromycin, itraconazole). Cinacalcet is a strong inhibitor of CYP2D6; therefore, dose adjustment of concomitant medications that are predominantly metabolized by CYP2D6 and have a narrow therapeutic index (e.g. flecainide, vinblastine, thioridazine and most tricyclic antidepressants) may be required. Cinacalcet does not appreciably inhibit or induce the activities of CYP3A4, 1A2, 2C9 or 2C19. An inverse relationship exists between plasma PTH and cinacalcet concentrations. PTH concentrations are greatest before dose administration when the cinacalcet concentration is lowest (24 hours after the previous day's dose). Nadir PTH levels occur approximately 2-3 hours after dosing. PMID- 19566114 TI - Characterization of the time course of carbamazepine deinduction by an enzyme turnover model. AB - BACKGROUND AND OBJECTIVE: Carbamazepine is a potent inducer of drug metabolizing enzymes, which results in a number of clinically significant drug-drug interactions. Deinduction occurs when long-term carbamazepine therapy is discontinued. The goal of this study was to develop a population pharmacokinetic model to describe the time course of carbamazepine deinduction. PATIENTS AND METHODS: Stable-labelled carbamazepine was administered intravenously on three occasions during the deinduction period to 15 patients with epilepsy for whom carbamazepine therapy was being discontinued. Data were analysed using a nonlinear mixed-effects model (NONMEM). An enzyme turnover model consisting of a one-compartment model linked with a hypothetical enzyme compartment was applied to characterize the time course of carbamazepine deinduction. Model evaluation was performed using the bootstrap approach and a visual predictive check. RESULTS: In the final model, the deinduction process was accomplished by decreasing the rate of enzyme synthesis, resulting in a decrease in the relative amount of enzymes. The estimated rate constant for enzyme degradation was 0.00805 h-1, corresponding to a half-life of the combined enzymes of 86.1 hours (3.6 days). CONCLUSION: An enzyme turnover model adequately characterized the experimental data. Based on the predicted enzyme half-life from the final model, the deinduction process should be completed within 2 weeks after carbamazepine therapy is terminated. PMID- 19566115 TI - Population pharmacokinetics of ganciclovir following administration of valganciclovir in paediatric renal transplant patients. AB - OBJECTIVE: To develop a population pharmacokinetic model for valganciclovir in paediatric renal transplant recipients, identify covariates that explain variability, and determine valganciclovir dosage regimens for cytomegalovirus (CMV) prophylaxis in children. METHODS: The pharmacokinetics of valganciclovir were described with plasma concentrations from 22 patients (age range 3-17 years) using nonlinear mixed-effects modelling (NONMEM) software. A two-compartment model with lag-time and first-order absorption and elimination was developed. The final model was validated using a bootstrap and visual predictive check. The dosage regimens of valganciclovir for CMV prophylaxis in children were simulated using the final model. RESULTS: The mean population pharmacokinetic parameters were apparent systemic clearance (CL) 10.1L/h, apparent central volume of distribution 5.2 L, apparent peripheral volume of distribution 30.7 L, inter tissue clearance 3.97 L/h, absorption rate constant 0.369 h-1 and lag-time 0.743 h. The covariate analysis identified creatinine clearance (CL(CR)) and bodyweight (WT) as individual factors influencing the apparent oral clearance: CL= 8.04 x (CL(CR)/89)(2.93) + 3.62 x (WT/28) L/h. The results of the simulation showed that for a typical patient (WT 28 kg and CL(CR) 89 mL/min), an area under the plasma concentration-time curve of 43 +/- 10.6 mg x h/mL will be achieved with valganciclovir 500 mg once daily. CONCLUSION: The dosage regimens of valganciclovir for CMV prophylaxis have been defined using the final population pharmacokinetic model based on WT and CLCR for paediatric renal transplantation patients. PMID- 19566117 TI - Therapeutic drug monitoring of aminoglycosides in neonates. PMID- 19566116 TI - Mycophenolic acid exposure after administration of mycophenolate mofetil in the presence and absence of cyclosporin in renal transplant recipients. AB - BACKGROUND AND OBJECTIVE: The pharmacokinetics of mycophenolic acid (MPA) are complex, with large interindividual variability over time. There are also well documented interactions with cyclosporin, and assessment of MPA exposure is therefore necessary when reducing or stopping cyclosporin therapy. Here we report on the pharmacokinetic and pharmacodynamic behaviour of MPA in renal transplant patients on standard dose, reduced dose and no cyclosporin. STUDY DESIGN: The CAESAR study, a prospective 12-month study in primary renal allograft recipients, was designed to determine whether mycophenolate mofetil-based regimens containing either low-dose cyclosporin or low-dose cyclosporin withdrawn by 6 months could minimize nephrotoxicity and improve renal function without an increase in acute rejection compared with a mycophenolate mofetil-based regimen containing standard dose cyclosporin. PATIENTS AND METHODS: A subset of patients from the CAESAR study contributed to this pharmacokinetic analysis of MPA exposure. Blood samples were taken over one dosing interval on day 7 and at months 3, 7 and 12 post transplantation. The sampling time points were predose, 20, 40 and 75 minutes and 2, 3, 4, 6, 8 and 12 hours after mycophenolate mofetil dosing. Assessments included plasma concentrations of MPA and mycophenolic acid glucuronide (MPAG) and cyclosporin trough concentrations. The area under the plasma concentration time curve (AUC) from 0 to 12 hours (AUC(12)) for MPA was the primary pharmacokinetic parameter, and the AUC(12) for MPAG was the secondary parameter. RESULTS: In total, 536 de novo renal allograft recipients were randomized in the CAESAR study. Of these, 114 patients were entered into the pharmacokinetic substudy and 110 patients contributed to the pharmacokinetic analysis. There was a rapid rise in MPA concentrations (median time to peak concentration 0.72-1.25 hours). At day 7 and month 3, the MPA AUC(12) values were similar in the cyclosporin withdrawal and low-dose cyclosporin groups (patients with the same cyclosporin target concentrations to month 6), while at 7 and 12 months, the values in the cyclosporin withdrawal group were higher than in the low-dose group (19.9% and 30.2% higher, respectively). MPA AUC(12) values in the standard-dose cyclosporin group were lower than in the other groups at all time points and increased over time. At all time points, the MPA peak plasma concentration was similar in all groups, and the MPAG concentrations rose more slowly than MPA concentrations. The ratio of the AUC from 6 to 12 hours/AUC(12) suggests that an increasing AUC in the cyclosporin withdrawal group is due to an increase in the enterohepatic recirculation. CONCLUSION: These findings are consistent with the hypothesis that cyclosporin inhibits the biliary secretion and/or hepatic extraction of MPAG, leading to a reduced rate of enterohepatic recirculation of MPA. Several concurrent mechanisms, such as cyclosporin-induced changes in renal tubular MPAG excretion and enhanced elimination of free MPA through competitive albumin binding with MPAG, can also contribute to the altered MPAG pharmacokinetics observed in the presence and absence of cyclosporin. PMID- 19566119 TI - HIV model incorporating differential progression for treatment-naive and treatment-experienced infectives. AB - We formulate an HIV/AIDS deterministic model which incorporates differential infectivity and disease progression for treatment-naive and treatment-experienced HIV/AIDS infectives. To illustrate our model, we have applied it to estimate adult HIV prevalence, the HIV population, the number of new infectives and the number of AIDS deaths for Botswana for the period 1984 to 2012. It is found that the prevalence peaked in the year 2000 and the HIV population is now decreasing. We have also found that under the current conditions, the reproduction number is Rc approximately 13, which is less than the 2004 estimate of Rc approximately equal 4 by [11] and [13]. The results in this study suggest that the HAART program has yielded positive results for Botswana. PMID- 19566120 TI - Blasting neuroblastoma using optimal control of chemotherapy. AB - A mathematical model is used to investigate the effectiveness of the chemotherapy drug Topotecan against neuroblastoma. Optimal control theory is applied to minimize the tumor volume and the amount of drug utilized. The model incorporates a state constraint that requires the level of circulating neutrophils (white blood cells that form an integral part of the immune system) to remain above an acceptable value. The treatment schedule is designed to simultaneously satisfy this constraint and achieve the best results in fighting the tumor. Existence and uniqueness of the solution of the optimality system, which is the state system coupled with the adjoint system, is established. Numerical simulations are given to demonstrate the behavior of the tumor and the immune system components represented in the model. PMID- 19566121 TI - Optimal control applied to vaccination and treatment strategies for various epidemiological models. AB - Mathematical models provide a powerful tool for investigating the dynamics and control of infectious diseases, but quantifying the underlying epidemic structure can be challenging especially for new and under-studied diseases. Variations of standard SIR, SIRS, and SEIR epidemiological models are considered to determine the sensitivity of these models to various parameter values that may not be fully known when the models are used to investigate emerging diseases. Optimal control theory is applied to suggest the most effective mitigation strategy to minimize the number of individuals who become infected in the course of an infection while efficiently balancing vaccination and treatment applied to the models with various cost scenarios. The optimal control simulations suggest that regardless of the particular epidemiological structure and of the comparative cost of mitigation strategies, vaccination, if available, would be a crucial piece of any intervention plan. PMID- 19566122 TI - Novel design of drug delivery in stented arteries: a numerical comparative study. AB - Implantation of drug eluting stents following percutaneous transluminal angioplasty has revealed a well established technique for treating occlusions caused by the atherosclerotic plaque. However, due to the risk of vascular re occlusion, other alternative therapeutic strategies of drug delivery are currently being investigated. Polymeric endoluminal pave stenting is an emerging technology for preventing blood erosion and for optimizing drug release. The classical and novel methodologies are compared through a mathematical model able to predict the evolution of the drug concentration in a cross-section of the wall. Though limited to an idealized configuration, the present model is shown to catch most of the relevant aspects of the drug dynamics in a delivery system. Results of numerical simulations shows that a bi-layer gel paved stenting guarantees a uniform drug elution and a prolonged perfusion of the tissues, and remains a promising and effective technique in drug delivery. PMID- 19566123 TI - Applications of occupancy urn models to epidemiology. AB - This paper shows how occupancy urn models can be used to derive useful results in epidemiology. First we show how simple epidemic models can be re-interpreted in terms of occupancy problems. We use this reformulation to derive an expression for the expected epidemic size, that is, the total number of infected at the end of an outbreak. We also use this approach to derive point and interval estimates of the Basic Reproduction Ratio, R0 . We show that this construction does not require that the underlying SIR model be a homogeneous Poisson process, leading to a geometric distribution for the number of contacts before removal, but instead it supports a general distribution. The urn model construction is easy to handle and represents a rich field for further exploitation. PMID- 19566125 TI - The dynamics of tumor growth and cells pattern morphology. AB - The mathematical modeling of tumor growth is an approach to explain the complex nature of these systems. A model that describes tumor growth was obtained by using a mesoscopic formalism and fractal dimension. This model theoretically predicts the relation between the morphology of the cell pattern and the mitosis/apoptosis quotient that helps to predict tumor growth from tumoral cells fractal dimension. The relation between the tumor macroscopic morphology and the cell pattern morphology is also determined. This could explain why the interface fractal dimension decreases with the increase of the cell pattern fractal dimension and consequently with the increase of the mitosis/apoptosis relation. Indexes to characterize tumoral cell proliferation and invasion capacities are proposed and used to predict the growth of different types of tumors. These indexes also show that the proliferation capacity is directly proportional to the invasion capacity. The proposed model assumes: i) only interface cells proliferate and invade the host, and ii) the fractal dimension of tumoral cell patterns, can reproduce the Gompertzian growth law. PMID- 19566126 TI - Finite-difference and pseudo-spectral methods for the numerical simulations of in vitro human tumor cell population kinetics. AB - Pseudo-spectral approximations are constructed for the model equations describing the population kinetics of human tumor cells in vitro and their responses to radiotherapy or chemotherapy. These approximations are more efficient than finite difference approximations. The spectral accuracy of the pseudo-spectral method allows us to resolve the model with a much smaller number of spatial grid-points than required for the finite-difference method to achieve comparable accuracy. This is demonstrated by numerical experiments which show a good agreement between predicted and experimental data. PMID- 19566127 TI - On the global dynamics of a model for tumor immunotherapy. AB - Understanding the dynamics of human hosts and tumors is of critical importance. A mathematical model was developed that explored the immune response to tumors that was used to study a special type of treatment. This treatment approach uses elements of the host to boost its immune response in the hopes that the host can clear the tumor. This model was extensively studied using numerical simulation, however no global analytical results were originally presented. In this work we explore the global dynamics to show under what conditions tumor clearance can be achieved. PMID- 19566128 TI - Global asymptotic properties for a Leslie-Gower food chain model. AB - We study global asymptotic properties of a continuous time Leslie-Gower food chain model. We construct a Lyapunov function which enables us to establish global asymptotic stability of the unique coexisting equilibrium state. PMID- 19566124 TI - A spatial model of tumor-host interaction: application of chemotherapy. AB - In this paper we consider chemotherapy in a spatial model of tumor growth. The model, which is of reaction-diffusion type, takes into account the complex interactions between the tumor and surrounding stromal cells by including densities of endothelial cells and the extra-cellular matrix. When no treatment is applied the model reproduces the typical dynamics of early tumor growth. The initially avascular tumor reaches a diffusion limited size of the order of millimeters and initiates angiogenesis through the release of vascular endothelial growth factor (VEGF) secreted by hypoxic cells in the core of the tumor. This stimulates endothelial cells to migrate towards the tumor and establishes a nutrient supply sufficient for sustained invasion. To this model we apply cytostatic treatment in the form of a VEGF-inhibitor, which reduces the proliferation and chemotaxis of endothelial cells. This treatment has the capability to reduce tumor mass, but more importantly, we were able to determine that inhibition of endothelial cell proliferation is the more important of the two cellular functions targeted by the drug. Further, we considered the application of a cytotoxic drug that targets proliferating tumor cells. The drug was treated as a diffusible substance entering the tissue from the blood vessels. Our results show that depending on the characteristics of the drug it can either reduce the tumor mass significantly or in fact accelerate the growth rate of the tumor. This result seems to be due to complicated interplay between the stromal and tumor cell types and highlights the importance of considering chemotherapy in a spatial context. PMID- 19566129 TI - The relative biologic effectiveness versus linear energy transfer curve as an output-input relation for linear cellular systems. AB - Experiments have established that different radiation types have different magnitudes of biological response. When biological response is defined in terms of the Relative Biologic Effectiveness (RBE) and different radiation type is characterized by Linear Energy Transfer (LET), the plot of the RBE versus LET (RBE-LET) curve shows RBE to increase with increasing LET, to reach a maximum, and to decrease with further increasing LET. Perhaps due to the descriptive nature of biology, most quantitative models for the RBE-LET curve ignore the reality of the underlying molecular biology. On the other hand, the molecular basis for the RBE-LET curve is not completely known despite recent efforts. Here we introduce a differential equation formulation for a signal-and-system model that sees cells as systems, different radiation types as input, and cellular responses as output. Because of scant knowledge of the underlying biochemical network, the current version is necessarily a work in progress. It explains the RBE-LET curve using not just input parameters but also systems internal state parameters. These systems internal state parameters represent parts of a biochemical network within a cell. Although multiple biochemical parts may well be involved, the shape of the RBE-LET curve is reproduced when only three system parameters are related to three biochemical parts: the molecular machinery for DNA double strand break repair; the molecular pathways for handling oxidative stress; and the radiolytic products of the cellular water. Despite being a simplified ''toy model,'' changes in the systems state parameters lead to model curves that are refutable in a modern molecular biology laboratory. As the parts in the biochemical network of the radiation response are being further elucidated, this model can incorporate new systems state parameters to allow a more accurate fit. PMID- 19566130 TI - Global stability for an SEIR epidemiological model with varying infectivity and infinite delay. AB - A recent paper (Math. Biosci. and Eng. (2008) 5:389-402) presented an SEIR model using an infinite delay to account for varying infectivity. The analysis in that paper did not resolve the global dynamics for R0 >1. Here, we show that the endemic equilibrium is globally stable for R0 >1. The proof uses a Lyapunov functional that includes an integral over all previous states. PMID- 19566131 TI - Theoretical modeling of RF ablation with internally cooled electrodes: comparative study of different thermal boundary conditions at the electrode tissue interface. AB - Previous studies on computer modeling of RF ablation with cooled electrodes modeled the internal cooling circuit by setting surface temperature at the coolant temperature (i.e., Dirichlet condition, DC). Our objective was to compare the temperature profiles computed from different thermal boundary conditions at the electrode-tissue interface. We built an analytical one-dimensional model based on a spherical electrode. Four cases were considered: A) DC with uniform initial condition, B) DC with pre-cooling period, C) Boundary condition based on Newton's cooling law (NC) with uniform initial condition, and D) NC with a pre cooling period. The results showed that for a long time (120 s), the profiles obtained with (Cases B and D) and without (Cases A and C) considering pre-cooling are very similar. However, for shorter times ( 30 s), Cases A and C overestimated the temperature at points away from the electrode-tissue interface. In the NC cases, this overestimation was more evident for higher values of the convective heat transfer coefficient (h). Finally, with NC, when h was increased the temperature profiles became more similar to those with DC. The results suggest that theoretical modeling of RF ablation with cooled electrodes should consider: 1) the modeling of a pre-cooling period, especially if one is interested in the thermal profiles registered at the beginning of RF application; and 2) NC rather than DC, especially for low flow in the internal circuit. PMID- 19566132 TI - Feedback stabilization for a chemostat with delayed output. AB - We apply basic tools of control theory to a chemostat model that describes the growth of one species of microorganisms that consume a limiting substrate. Under the assumption that available measurements of the model have fixed delay t>0, we design a family of feedback control laws with the objective of stabilizing the limiting substrate concentration in a fixed level. Effectiveness of this control problem is equivalent to global attractivity of a family of differential delay equations. We obtain sufficient conditions (upper bound for delay t>0 and properties of the feedback control) ensuring global attractivity and local stability. Illustrative examples are included. PMID- 19566133 TI - A model for transmission of partial resistance to anti-malarial drugs. AB - Anti-malarial drug resistance has been identified in many regions for a long time. In this paper we formulate a mathematical model of the spread of anti malarial drug resistance in the population. The model is suitable for malarial situations in developing countries. We consider the sensitive and resistant strains of malaria. There are two basic reproduction ratios corresponding to the strains. If the ratios corresponding to the infections of the sensitive and resistant strains are not equal and they are greater than one, then there exist two endemic non-coexistent equilibria. In the case where the two ratios are equal and they are greater than one, the coexistence of the sensitive and resistant strains exist in the population. It is shown here that the recovery rates of the infected host and the proportion of anti-malarial drug treatment play important roles in the spread of anti-malarial drug resistance. The interesting phenomena of ''long-time" coexistence, which may explain the real situation in the field, could occur for long period of time when those parameters satisfy certain conditions. In regards to control strategy in the field, these results could give a good understanding of means of slowing down the spread of anti-malarial drug resistance. PMID- 19566134 TI - New developments in using stochastic recipe for multi-compartment model: inter compartment traveling route, residence time, and exponential convolution expansion. AB - Drug residence time in ''compartmentalized'' human body system had been studied from both deterministic and Markovian perspectives. However, probability and probability density functions for a drug molecule to be (1) in any compartment of study interest, (2) with any defined inter-compartment traveling route, and (3) with/without specified residence times in its visited compartments, has not been systemically reported. In Markovian view of compartmental system, mathematical solutions for the probability or probability density functions, for a drug molecule with any defined inter- compartment traveling routes in the system and/or with specified residence times in any visited compartments, are provided. Matrix convolution is defined and thus employed to facilitate methodology development. Laplace transformations are used to facilitate convolution operations in linear systems. This paper shows that the drug time-concentration function can be decomposed into the summation of a series of component functions, which is named as convolution expansion. The studied probability or probability density functions can be potentially engaged with physiological or pharmacological significances and thus be used to describe a broad range of drug exposure-response relationships. PMID- 19566135 TI - Local explicitly correlated coupled-cluster methods: efficient removal of the basis set incompleteness and domain errors. AB - We propose an explicitly correlated local LCCSD-F12 method in which the basis set incompleteness error as well as the error caused by truncating the virtual orbital space to pair-specific local domains are strongly reduced. This is made possible by including explicitly correlated terms that are orthogonalized only to the pair-specific configuration space. Thus, the contributions of excitations outside the domains are implicitly accounted for by the explicitly correlated terms. It is demonstrated for a set of 54 reactions that the reaction energies computed with the new LCCSD-F12 method and triple-zeta basis sets deviate by at most 2.5 kJ/mol from conventional CCSD complete basis set results (RMS: 0.6 kJ/mol). The local approximations should make it possible to achieve linear scaling of the computational cost with molecular size. PMID- 19566136 TI - Quantum mechanics with the basis set guided by Ehrenfest trajectories: theory and application to spin-boson model. AB - In this article a method of numerical solution of the Schrodinger equation is proposed. The approach corrects the Ehrenfest approximation by using several trajectories/configurations with their amplitudes coupled within and across configurations, thus making the method formally exact. Accurate results are obtained for the spin-boson model with up to 2000 bath modes treated on fully quantum level without approximations. PMID- 19566137 TI - Relativistic, QED, and nuclear mass effects in the magnetic shielding of 3He. AB - The magnetic shielding sigma of (3)He is studied. The complete relativistic corrections of order O(alpha(2)), leading QED corrections of order O(alpha(3) ln alpha), and finite nuclear mass effects of order O(m/m(N)) are calculated with high numerical precision. The resulting theoretical predictions for sigma = 59.967 43(10)x10(-6) are the most accurate to date among all elements and support the use of (3)He as a NMR standard. PMID- 19566138 TI - Variable Lieb-Oxford bound satisfaction in a generalized gradient exchange correlation functional. AB - We propose a different way to satisfy both gradient expansion limiting behavior and the Lieb-Oxford bound in a generalized gradient approximation exchange functional by extension of the Perdew-Burke-Ernzerhof (PBE) form. Motivation includes early and recent exploration of modified values for the gradient expansion coefficient in the PBE exchange-correlation functional (cf. the PBEsol functional) and earlier experience with a numerical cutoff for large-s (s proportional to absolute value(vector differential n)/n(4/3)) in a version of the deMon molecular code. For either the original PBE or the PBEsol choice of the gradient coefficient, we find improved performance from using an s-dependent (spatially varying) satisfaction of the Lieb-Oxford bound which quenches to uniform electron gas behavior at large s. The mean absolute deviations (MADs) in atomization energies for a widely used test set of 20 small molecules are reduced by about 22% relative to PBE and PBEsol. For these small molecules, the bond length MADs are essentially unchanged. PMID- 19566140 TI - Relationships between the third-order reactivity indicators in chemical density functional theory. AB - Relationships between third-order reactivity indicators in the closed system [N, v(r)], open system [mu, v(r)], and density [rho(r)] pictures are derived. Our method of derivation unifies and extends known results. Among the relationships is a link between the third-order response of the energy to changes in the density and the quadratic response of the density to changes in external potential. This provides a link between hyperpolarizability and the system's sensitivity to changes in electron density. The dual descriptor is a unifying feature of many of the formulas we derive. PMID- 19566139 TI - A new class of highly efficient exact stochastic simulation algorithms for chemical reaction networks. AB - We introduce an alternative formulation of the exact stochastic simulation algorithm (SSA) for sampling trajectories of the chemical master equation for a well-stirred system of coupled chemical reactions. Our formulation is based on factored-out, partial reaction propensities. This novel exact SSA, called the partial-propensity direct method (PDM), is highly efficient and has a computational cost that scales at most linearly with the number of chemical species, irrespective of the degree of coupling of the reaction network. In addition, we propose a sorting variant, SPDM, which is especially efficient for multiscale reaction networks. PMID- 19566141 TI - From power law intermittence to macroscopic coherent regime. AB - We address the problem of establishing which is the proper form of quantum master equation generating a survival probability identical to that corresponding to the nonergodic sequence of "light on" and "light off" fluorescence fluctuations in blinking quantum dots. We adopt a theoretical perspective based on the assumption that the abrupt transitions from the light on to light off state are the results of many collisions between system and environment, properly described by the Lindblad equation, and that between two consecutive collisions the system dynamics are frozen. This generates a quantum master equation belonging to the recently proposed class of generalized Lindblad equations, with a time convoluted structure, involving in the specific case of this paper both the unitary and the nonunitary contribution of the Lindblad equation. This is the property that under the low-frequency condition makes the new class of generalized Lindblad equation generates the required survival probability. We make the conjecture that this equation corresponds to the cooperative dynamics of many units that, in isolation, are described by the ordinary Lindblad equation. When the time scale of the unitary term of the Lindblad equation is shorter than the dephasing time, the cooperation generates a surprisingly extended macroscopic coherence. PMID- 19566142 TI - Decoherence and dissipation in a molecular system coupled to an environment: an application of semiclassical hybrid dynamics. AB - Applying the recently developed semiclassical hybrid dynamics [Grossmann, J. Chem. Phys. 125, 014111 (2006)], we study the decay of interference patterns in the reduced density as well as of the purity in a Morse oscillator test system due to the interaction with a finite harmonic bath at zero temperature. In the case that the bath mimics a continuous Ohmic spectral density, in addition to the quantum classical transition induced by the interaction with the environment, we corroborate the existence of a blueshift due to the bath coupling, predicted by Pollak [Phys. Rev. A 33, 4244 (1986)]. Furthermore, the decoherence dynamics of cat states is confirmed to be faster than that of single coherent states and we show that for a resonant bath the dissipation leads to an increase in the decoherence rate as compared to the low frequency bath. PMID- 19566143 TI - Transition state-finding strategies for use with the growing string method. AB - Efficient identification of transition states is important for understanding reaction mechanisms. Most transition state search algorithms require long computational times and a good estimate of the transition state structure in order to converge, particularly for complex reaction systems. The growing string method (GSM) [B. Peters et al., J. Chem. Phys. 120, 7877 (2004)] does not require an initial guess of the transition state; however, the calculation is still computationally intensive due to repeated calls to the quantum mechanics code. Recent modifications to the GSM [A. Goodrow et al., J. Chem. Phys. 129, 174109 (2008)] have reduced the total computational time for converging to a transition state by a factor of 2 to 3. In this work, three transition state-finding strategies have been developed to complement the speedup of the modified-GSM: (1) a hybrid strategy, (2) an energy-weighted strategy, and (3) a substring strategy. The hybrid strategy initiates the string calculation at a low level of theory (HF/STO-3G), which is then refined at a higher level of theory (B3LYP/6-31G(*)). The energy-weighted strategy spaces points along the reaction pathway based on the energy at those points, leading to a higher density of points where the energy is highest and finer resolution of the transition state. The substring strategy is similar to the hybrid strategy, but only a portion of the low-level string is refined using a higher level of theory. These three strategies have been used with the modified-GSM and are compared in three reactions: alanine dipeptide isomerization, H-abstraction in methanol oxidation on VO(x)/SiO(2) catalysts, and C-H bond activation in the oxidative carbonylation of toluene to p toluic acid on Rh(CO)(2)(TFA)(3) catalysts. In each of these examples, the substring strategy was proved most effective by obtaining a better estimate of the transition state structure and reducing the total computational time by a factor of 2 to 3 compared to the modified-GSM. The applicability of the substring strategy has been extended to three additional examples: cyclopropane rearrangement to propylene, isomerization of methylcyclopropane to four different stereoisomers, and the bimolecular Diels-Alder condensation of 1,3-butadiene and ethylene to cyclohexene. Thus, the substring strategy used in combination with the modified-GSM has been demonstrated to be an efficient transition state finding strategy for a wide range of types of reactions. PMID- 19566144 TI - Phase equilibria of molecular fluids via hybrid Monte Carlo Wang-Landau simulations: applications to benzene and n-alkanes. AB - In recent years, powerful and accurate methods, based on a Wang-Landau sampling, have been developed to determine phase equilibria. However, while these methods have been extensively applied to study the phase behavior of model fluids, they have yet to be applied to molecular systems. In this work, we show how, by combining hybrid Monte Carlo simulations in the isothermal-isobaric ensemble with the Wang-Landau sampling method, we determine the vapor-liquid equilibria of various molecular fluids. More specifically, we present results obtained on rigid molecules, such as benzene, as well as on flexible chains of n-alkanes. The reliability of the method introduced in this work is assessed by demonstrating that our results are in excellent agreement with the results obtained in previous work on simple fluids, using either transition matrix or conventional Monte Carlo simulations with a Wang-Landau sampling, and on molecular fluids, using histogram reweighting or Gibbs ensemble Monte Carlo simulations. PMID- 19566145 TI - Electronic structure and molecular dynamics of breaking the RO-NO2 bond. AB - Decomposition of energetic molecules such as pentaerythritol tetranitrate is accompanied by extensive changes in their electronic configuration and thus is challenging for ab initio Born-Oppenheimer molecular dynamics simulations. The performance of single-determinant methods (in particular, density-functional theory) is validated on electronic structure and molecular dynamics simulations of RO-NO(2) bond dissociation in a smaller nitric ester, ethyl nitrate. Accurate description of dissociating molecule requires using unrestricted, spin-symmetry broken orbitals. However, the iterative self-consistent field procedure is prone to convergence failures in the bond-breaking region even if robust convergence algorithms are employed. As a result, molecular dynamics simulations of unimolecular decomposition need to be closely monitored and manually restarted to ensure seamless transition from the closed-shell to open-shell configuration. PMID- 19566146 TI - Analytic dynamics of the Morse oscillator derived by semiclassical closures. AB - The quantized Hamilton dynamics methodology [O. V. Prezhdo and Y. V. Pereverzev, J. Chem. Phys. 113, 6557 (2000)] is applied to the dynamics of the Morse potential using the SU(2) ladder operators. A number of closed analytic approximations are derived in the Heisenberg representation by performing semiclassical closures and using both exact and approximate correspondence between the ladder and position-momentum variables. In particular, analytic solutions are given for the exact classical dynamics of the Morse potential as well as a second-order semiclassical approximation to the quantum dynamics. The analytic approximations are illustrated with the O-H stretch of water and a Xe-Xe dimer. The results are extended further to coupled Morse oscillators representing a linear triatomic molecule. The reported analytic expressions can be used to accelerate classical molecular dynamics simulations of systems containing Morse interactions and to capture quantum-mechanical effects. PMID- 19566147 TI - Threshold photoionization and density functional theory studies of bimetallic carbide nanocrystals and fragments: Ta3ZrC(y) (y = 0-4). AB - Gas-phase bimetallic tantalum-zirconium-carbide clusters are generated using a constructed double ablation cluster source. The Ta(3)ZrC(y) (y = 0-4) clusters are examined by photoionization efficiency spectroscopy to extract experimental ionization energies (IEs). The IE trend for the Ta(3)ZrC(y) cluster series is reasonably similar to that of the Ta(4)C(y) cluster series [V. Dryza et al., J. Phys. Chem. A 109, 11180 (2005)], although the IE reductions upon carbon addition are greater for the former. Complementary density functional theory calculations are performed for the various isomers constructed by attaching carbon atoms to the different faces of the tetrahedral Ta(3)Zr cluster. The good agreement between the experimental IE trend and that calculated for these isomers support a 2x2x2 face centered cubic nanocrystal structure for Ta(4)ZrC(4) and nanocrystal fragment structures for the smaller clusters. PMID- 19566148 TI - The dynamics of the H2 + CO+ reaction on an interpolated potential energy surface. AB - A potential energy surface that describes the title reaction has been constructed by interpolation of ab initio data. Classical trajectory studies on this surface show that the total reaction rate is close to that predicted by a Langevin model, although the mechanism is more complicated than simple ion-molecule capture. Only the HCO(+) + H product is observed classically. An estimate of the magnitude of rotational inelastic scattering is also reported. PMID- 19566149 TI - On the accuracy of thermionic electron emission models. I. Electron detachment from SF6(-). AB - Detailed statistical rate calculations combined with electron capture theory and kinetic modeling for the electron attachment to SF(6) and detachment from SF(6)( ) [Troe et al., J. Chem. Phys. 127, 244303 (2007)] are used to test thermionic electron emission models. A new method to calculate the specific detachment rate constants k(det)(E) and the electron energy distributions f(E,epsilon) as functions of the total energy E of the anion and the energy epsilon of the emitted electrons is presented, which is computationally simple but neglects fine structures in the detailed k(det)(E). Reduced electron energy distributions f(E,epsilon/) were found to be of the form (epsilon/)(n) exp( epsilon/) with n approximately = 0.15, whose shape corresponds to thermal distributions only to a limited extent. In contrast, the average energies can be roughly estimated within thermionic emission and finite heat bath concepts. An effective temperature T(d)(E) is determined from the relation E - EA = + kT(d), where denotes the thermal internal energy of the detachment product SF(6) at the temperature T(d) and EA is the electron affinity of SF(6). The average electron energy is then approximately given by = kT(d)(E), but dynamical details of the process are not accounted for by this approach. Simplified representations of k(det)(E) in terms of T(d)(E) from the literature are shown to lead to only semiquantitative agreement with the equally simple but more accurate calculations presented here. An effective "isokinetic" electron emission temperature T(e)(E) does not appear to be useful for the electron detachment system considered because it neither provides advantages over a representation of k(det)(E) as a function of T(d)(E), nor are recommended relations between T(e)(E) and T(d)(E) of sufficient accuracy. PMID- 19566150 TI - Correction for dispersion and Coulombic interactions in molecular clusters with density functional derived methods: application to polycyclic aromatic hydrocarbon clusters. AB - The density functional based tight binding (DFTB) is a semiempirical method derived from the density functional theory (DFT). It inherits therefore its problems in treating van der Waals clusters. A major error comes from dispersion forces, which are poorly described by commonly used DFT functionals, but which can be accounted for by an a posteriori treatment DFT-D. This correction is used for DFTB. The self-consistent charge (SCC) DFTB is built on Mulliken charges which are known to give a poor representation of Coulombic intermolecular potential. We propose to calculate this potential using the class IV/charge model 3 definition of atomic charges. The self-consistent calculation of these charges is introduced in the SCC procedure and corresponding nuclear forces are derived. Benzene dimer is then studied as a benchmark system with this corrected DFTB (c DFTB-D) method, but also, for comparison, with the DFT-D. Both methods give similar results and are in agreement with references calculations (CCSD(T) and symmetry adapted perturbation theory) calculations. As a first application, pyrene dimer is studied with the c-DFTB-D and DFT-D methods. For coronene clusters, only the c-DFTB-D approach is used, which finds the sandwich configurations to be more stable than the T-shaped ones. PMID- 19566151 TI - Ab initio study of methyl-bromide photodissociation in the A band. AB - We performed a theoretical study of the photodissociation dynamics of CH(3)Br in the A band using a wave packet propagation technique on coupled ab initio potential energy curves. The present model involves the (3)Q(1) and (1)Q(1) excited states which can be populated from the ground state by a perpendicular transition and which are correlated at large methyl-bromide distance to the ground bromide spin-orbit state, as well as the (3)Q(0) and 4E states which can be excited by a parallel and perpendicular transition (respectively) and both correlate to excited Br(*) spin-orbit state. The model provides absorption cross sections and branching ratios in excellent agreement with experimental results. Due to weak spin-orbit interaction, the (1)Q(1) state is the dominant contributor to the absorption cross section, except for the red wing of the band where (3)Q(0) and (3)Q(1) states have significant absorption. However, spin-orbit coupling is strong enough to induce nonadiabatic transitions between the (3)Q(1) and (1)Q(1) states during the dissociation process which should be experimentally detectable in the alignment properties of the fragments. Nonadiabatic transitions at the conical intersection between (3)Q(0) and (1)Q(1) are shown to play a minor role in this system. PMID- 19566152 TI - Ab initio study of valence and Rydberg states of CH3Br. AB - We performed configuration interaction ab initio calculations on the valence and 5s, 5p(a(1)), and 5p(e) Rydberg bands of the CH(3)Br molecule as a function of the methyl-bromide distance for frozen C(3v) geometries. The valence state potential energy curves are repulsive, the Rydberg state ones are similar to the one of the CH(3)Br(+) ion with a minimum at short distance. One state emerging from the 5p(e) band has valence and ion-pair characters as distance increases and the corresponding potential curve has a second minimum at large distance. This state has a very strong parallel electric dipole transition moment with the ground state and plays a central role in UV photon absorption spectra. It is also responsible for the parallel character of the anisotropy parameters measured in ion-pair production experiments. In each band, there is a single state, which has a non-negligible transition moment with the ground state, corresponding to a transition perpendicular to the molecular axis of symmetry, except for the 5p(e) band where it is parallel. The perpendicular transition moments between ground and valence states increase sharply as methyl-bromide distance decreases due to a mixing between valence and 5s Rydberg band at short distance. In each band, spin orbit interaction produces a pair of states, which have significant transition moments with the ground one. In the valence band, the mixing between singlet and triplet states is weak and the perpendicular transition to the (1)Q(1) state is dominant. In each Rydberg band, however, spin-orbit interaction is larger than the exchange interaction and the two significant transition moments with the ground state have comparable strengths. The valence band has an additional state ((1)Q(0)) with significant parallel transition moment induced by spin-orbit interaction with the ground state at large distance. PMID- 19566153 TI - Theoretical investigation of the Omega(g,u)(+/-) states of K2 dissociating adiabatically up to K(4p 2P(3/2)) + K(4p 2P(3/2)). AB - A theoretical investigation of the electronic structure of the K(2) molecule, including spin-orbit effects, has been performed. Potential energies have been calculated over a large range of R up to 75a(0) for the 88 Omega(g,u)(+/-) states dissociating adiabatically into the limits up to K(4p (2)P(3/2))+K(4p (2)P(3/2)). Equilibrium distances, transition energies, harmonic frequencies, as well as depths for wells and heights for barriers are reported for all of the bound Omega(g,u)(+/-) states. Present ab initio calculations are shown to be able to reproduce quite accurately the small structures (wells and barrier) displayed at very long-range (R>50a(0)) by the (2,3)1(u) and (2)0(g)(-) purely long-range states. As the present data could help experimentalists, we make available extensive tables of energy values versus internuclear distances in our database at the web address http://www-lasim.univ-lyon1.fr/spip.php?rubrique99. PMID- 19566154 TI - The mechanism of the interstellar isomerization reaction HOC+ --> HCO+ catalyzed by H2: new insights from the reaction electronic flux. AB - A theoretical study of the mechanism of the isomerization reaction HOC(+) --> HCO(+) is presented. The mechanism was studied in terms of reaction force, chemical potential, reaction electronic flux (REF), and bond orders. It has been found that the evolution of changes in REF along the intrinsic reaction coordinate can be explained in terms of bond orders. The energetic lowering of the hydrogen assisted (catalyzed) reaction has been identified as being due to the stabilization of the H(3)(+) transition state complex and the stepwise bond dissociation and formation of the H-O and H-C bonds, respectively. PMID- 19566155 TI - Slow photoelectron velocity-map imaging spectroscopy of the vinoxide anion. AB - High resolution photoelectron spectra of the vinoxide anion are obtained by slow electron velocity-map imaging. Transitions between the anion X (1)A(') ground electronic state and the radical X (2)A(") and A (2)A(') states are observed. This experiment yields a precise value of 1.8250+/-0.0012 eV for the adiabatic electron affinity and 0.996+/-0.003 eV for the A-X term energy of the vinoxy radical. Franck-Condon simulations of the X (2)A(") <-- X (1)A(') transition are performed at varying levels of approximation. Full treatment with Duschinsky rotation is necessary to reproduce experimental results. Comparison of the experimental and simulated spectra leads to the assignment of previously unresolved transitions, notably between levels of a(") symmetry. PMID- 19566156 TI - An exchange-Coulomb model potential energy surface for the Ne-CO interaction. I. Calculation of Ne-CO van der Waals spectra. AB - Exchange-Coulomb model potential energy surfaces have been developed for the Ne CO interaction. The initial model is a three-dimensional potential energy surface based upon computed Heitler-London interaction energies and literature results for the long-range induction and dispersion energies, all as functions of interspecies distance, the orientation of CO relative to the interspecies axis, and the bond length of the CO molecule. Both a rigid-rotor model potential energy surface, obtained by setting the CO bond length equal to its experimental spectroscopic equilibrium value, and a vibrationally averaged model potential energy surface, obtained by averaging the stretching dependence over the ground vibrational motion of the CO molecule, have been constructed from the full data set. Adjustable parameters in each model potential energy surface have been determined through fitting a selected subset of pure rotational transition frequencies calculated for the (20)Ne-(12)C(12)O isotopolog to precisely known experimental values. Both potential energy surfaces provide calculated results for a wide range of available experimental microwave, millimeter-wave, and midinfrared Ne-CO transition frequencies that are generally far superior to those obtained using the best current literature potential energy surfaces. The vibrationally averaged CO ground state potential energy surface, employed together with a potential energy surface obtained from it by replacing the ground vibrational state average of the CO stretching dependence of the potential energy surface by an average over the first excited CO vibrational state, has been found to be particularly useful for computing and/or interpreting mid-IR transition frequencies in the Ne-CO dimer. PMID- 19566157 TI - Temperature dependence of reactions involving electron transfer in K(np)/C2Cl4 collisions. AB - Electron transfer in K(np)-C(2)Cl(4) collisions, which leads to formation of both Cl(-) and C(2)Cl(4)(-) anions, is investigated as a function of target temperature over the range of 300-650 K. Measurements at high n (n approximately 30) show that the likelihood of Cl(-) production increases rapidly with temperature indicating the presence of a dissociation barrier. The data yield an activation energy of approximately 0.1 eV. A broad distribution of product C(2)Cl(4)(-) lifetimes is observed that extends from microseconds to milliseconds, this distribution moving toward shorter lifetimes as the target temperature is increased. The measured lifetimes are consistent with the predictions of quasiequilibrium theory. Studies at low n (n approximately 14) show a substantial fraction of the product K(+)-Cl(-) and K(+)-C(2)Cl(4)(-) ion pairs is electrostatically bound leading to creation of heavy-Rydberg ion-pair states. Variations in target temperature lead to changes in kinetic energy of relative motion of the reactants that can result in marked changes in the fraction of ion pairs that is bound, especially at low Rydberg atom velocities. In the case of bound K(+)-C(2)Cl(4)(-) ion pairs a few percent subsequently dissociate by the conversion of internal energy in the anion into translational energy of the ion pair. Analysis of the data points to a mean energy conversion of approximately 60-90 meV, much less than the available excess energy of reaction, approximately 0.7 eV. PMID- 19566158 TI - Vibration energy levels of the PH3, PH2D, and PHD2 molecules calculated from high order potential energy surface. AB - Vibrational energy levels of the PH(3), PH(2)D, and PHD(2) molecules were calculated from the new extended potential energy surface (PES) determined in this work. The coupled-cluster approach with the perturbative inclusion of the connected triple excitations CCSD(T) and correlation consistent polarized valence basis set cc-pV5Z was employed in the ab initio calculations of electronic ground state energies. The contribution of relativistic effects to the overall electronic energy surface was computed using quasirelativistic mass-velocity Darwin approach. These ab initio points were fitted by a parametrized function with one parameter empirically adjusted. The grid of 11,697 geometrical nuclear configurations covers a large domain of the six dimensional internal coordinate space and was designed to provide vibration energy levels of phosphine molecule up to 7000 cm(-1) above the zero point vibration energy with reasonable accuracy. The analytical representation of the PES was determined through the expansion in symmetry adapted products of nonlinear internal coordinates for various orders of analytical expansions up to the tenth order. The dependence of calculated vibration energy levels on the analytical representation of PES and on the coordinate choice was studied. Calculated vibration levels are in very good agreement with observations: The root mean squares deviation between theoretically calculated and observed band centers is 1.4 cm(-1) for PH(3), 0.4 cm(-1) for PH(2)D, and 0.6 cm(-1) for PHD(2). PMID- 19566159 TI - Chirped pulse multiphoton ionization of nitrogen: control of selective rotational excitation in N(2)(+)(B 2Sigma(u)(+)). AB - We report on fluorescence spectra of N(2)(+)(B (2)Sigma(u)(+)) --> N(2)(+)(X (2)Sigma(g)(+)) obtained from multiphoton ionization of molecular nitrogen by 804 nm femtosecond laser pulses. The analysis of the fluorescence spectra reveals that the vibrational levels v = 0 and v = 4 in the B (2)Sigma(u)(+)-state of N(2)(+) are primarily populated. The rotational state distribution of N(2)(+)(B (2)Sigma(u)(+), v = 0) is determined from the rotationally resolved fluorescence spectra. It is demonstrated that the linear chirp of the 804 nm femtosecond laser pulse has a strong influence on the rotational state distribution of the vibrational ground state of the molecular cation N(2)(+)(B (2)Sigma(u)(+), v = 0). Possible mechanisms leading to the experimental results are discussed. The particular population of the vibrational levels as well as the linear chirp dependence of the fluorescence signal gives evidence for the importance of a resonant intermediate state. The N(2) a (1)Pi-state is likely involved in a resonant multiphoton excitation process. This permits to selectively control the rotational population of the cation that is formed via chirped pulse multiphoton ionization. PMID- 19566160 TI - Nonequilibrium Green's function study on the electronic structure and transportation behavior of the conjugated molecular junction: terminal connections and intramolecular connections. AB - In the recent density functional-based calculations, it was found that the conductivity of naphthalene molecular wires can be modulated by altering the linking position of the molecule to the electrode [D. Walter, D. Neuhauser, and R. Baer, Chem. Phys. 299, 139 (2004)]. A quantum interference model was proposed to interpret the observation. In this paper, we further studied the conductance of a series of conjugated molecules containing aromatic rings using density functional theory combined with nonequilibrium Green's function method. For polyacene systems with different terminal connections, the conductivity is dependent on the substitution position of anchoring groups even with similar electron transport distance. The conductance of trans-substitution can be ten times or more as large as that of the cis-substitution. However, for the biphenyl system with different intramolecular connections, adding more connections between two benzene rings does not change the junction conductance. All these results indicate that the junction conductance is strongly dependent on the particular electron transport pathway. The alternating double-single linkage is the most probable one, since others are impeded by the single bonds. PMID- 19566161 TI - Photon counting statistics of single molecule in solid matrix. AB - In this paper, we investigate the properties of photon emission statistics of single molecule in solid matrix. The influences of solid matrix surroundings on photon emission of single molecule system under the laser field and rf field for several examples, the single dibenzanthanthrene molecule in hexadecane, the spectral diffusion process, and the hidden two-state models and the Gaussian models of blinking behavior, are considered. PMID- 19566162 TI - Predicted bound states and microwave spectrum of N2-He van der Waals complexes. AB - Numerical calculations show that four modern potential energy surfaces for N(2) He all support 18 bound intermolecular states for the homonuclear isotopologues (14,14)N(2)-(4)He and (15,15)N(2)-(4)He, and 12 (or 13, for one surface) truly bound states for (14,15)N(2)-He. This contradicts a recent statement [Patel et al., J. Chem. Phys. 119, 909 (2003)] that one of these surfaces supports no bound states, and it yields predictions for 27 allowed pure rotational transitions among the truly bound states of the homonuclear isotopologues of this complex. PMID- 19566163 TI - Plastic crystal phases of simple water models. AB - We report the appearance of two plastic crystal phases of water at high pressure and temperature using computer simulations. In one of them the oxygen atoms form a body centered cubic structure (bcc) and in the other they form a face centered cubic structure (fcc). In both cases the water molecules were able to rotate almost freely. We have found that the bcc plastic crystal transformed into a fcc plastic crystal via a Martensitic phase transition when heated at constant pressure. We have performed the characterization and localization in the phase diagram of these plastic crystal phases for the SPC/E, TIP4P, and TIP4P/2005 water potential models. For TIP4P/2005 model free energy calculations were carried out for the bcc plastic crystal and fcc plastic crystal using a new method (which is a slight variation of the Einstein crystal method) proposed for these types of solid. The initial coexistence points for the SPC/E and TIP4P models were obtained using Hamiltonian Gibbs-Duhem integration. For all of these models these two plastic crystal phases appear in the high pressure and temperature region of the phase diagram. It would be of interest to study if such plastic crystal phases do indeed exist for real water. This would shed some light on the question of whether these models can describe satisfactorily the high pressure part of the phase diagram of water, and if not, where and why they fail. PMID- 19566164 TI - Growing correlation length in supercooled water. AB - The evolution of the structure of water from the stable high temperature liquid to its glass, low-density amorphous ice (LDA), is studied through large-scale molecular dynamics simulations with the mW model [J. Phys. Chem. B 113, 4008 (2009)]. We characterize the density, translational, and orientational ordering of liquid water from the high temperature stable liquid to the low-density glass LDA at the critical cooling rate for vitrification. A continuous transition to a tetrahedrally ordered low-density liquid is observed at 50 K below the temperature of maximum density and 25 K above a temperature of minimum density. The structures of the low-density liquid and glass are consistent with that of a continuous random tetrahedral network. The liquid-liquid transformation temperature T(LL), defined by the maximum isobaric expansivity, coincides with the maximum rate of change in the local structure of water. Long-range structural fluctuations of patches of four-coordinated molecules form in the liquid. The correlation length of the four-coordinated patches in the liquid increases according to a power law in the range 300 K to T(LL)+10 K; a maximum is predicted at T(LL). To the best of our knowledge this is the first direct estimation of the Widom line of supercooled water through the analysis of structural correlations. PMID- 19566165 TI - Effects of the mean free path and relaxation in a model for the aggregation of particles in superfluid media. AB - In this paper, we study a two-dimensional model for the growth of molecular clusters in superfluid helium at low temperature. In the model, particles of diameter a follow random ballistic moves of length delta = a - 256a. Upon attachment on the cluster surface, particles allow one-step relaxation to the nearest twofold coordinated site. Average coordination numbers of particles show that in the presence of relaxation the screening for incoming particles modifies the microscopic structure of the cluster. These results are in qualitative agreement with experimental aggregation of methane in He droplets, in which large abundance of fully coordinated sites is observed. The average coordination number increases with delta, showing that screening is relevant when relaxation is present. As the cluster size increases, the corresponding structure clusters changes from a compact to a fractal, typical of ballistic and diffusion limited models, respectively. A scaling ansatz describing the crossover between the two regimes is proposed. PMID- 19566166 TI - Minimum free-energy path of homogenous nucleation from the phase-field equation. AB - The minimum free-energy path (MFEP) is the most probable route of the nucleation process on the multidimensional free-energy surface. In this study, the phase field equation is used as a mathematical tool to deduce the MFEP of homogeneous nucleation. We use a simple square-gradient free-energy functional with a quartic local free-energy function as an example and study the time evolution of a single nucleus placed within a metastable environment. The time integration of the phase field equation is performed using the numerically efficient cell-dynamics method. By monitoring the evolution of the size of the nucleus and the free energy of the system simultaneously, we can easily deduce the free-energy barrier as a function of the size of the sub- and the supercritical nucleus along the MFEP. PMID- 19566167 TI - Crystal field theory analysis of rovibrational spectra of carbon monoxide monomers isolated in solid parahydrogen. AB - We report the first rotationally resolved and completely assigned rovibrational spectrum for a nonhydride molecule rotating in the solid phase: carbon monoxide (CO) monomers isolated in cryogenic solid parahydrogen (p-H(2)). We employ a modified crystal field theory model, in which the CO molecular spectroscopic constants are taken as adjustable parameters, to make good spectroscopic assignments for all the observed features. We discuss the limitations of this approach and highlight the need for improved theoretical models of molecular rotation dynamics in quantum solids. PMID- 19566168 TI - A molecular dynamics study of chirality transfer from chiral surfaces to nearby solvent. AB - The presence of a chiral surface can alter the characteristics of nearby solvent molecules such that, on average, these molecules become chiral. The extent of this induced chirality and its dependence on the surface and solvent characteristics are explored in this article. Three surfaces employed in chiral chromatography are examined: The Whelk-O1 interface, a phenylglycine-derived chiral stationary phase (CSP), and a leucine-derived CSP. All three interfaces are "brush type" in that the chiral molecules are attached to the underlying substrate via an achiral tether. The solvents consist of ethanol, a binary n hexane/ethanol solvent, 2-propanol, and a binary n-hexane/2-propanol solvent. Molecular dynamics simulations of the solvated chiral interfaces form the basis of the analysis. The chirality induced in the solvent is assessed based on a chirality index originally proposed by Osipov et al. [Mol. Phys. 84, 1193 (1995)]. Solvent chirality will depend on the solvent position relative to the surface. For this reason, a position-dependent chirality index is analyzed in detail. PMID- 19566169 TI - Mesoscopic simulations of the counterion-induced electro-osmotic flow: a comparative study. AB - We present mesoscopic simulations of the counterion-induced electro-osmotic flow in different electrostatic coupling regimes. Two simulation methods are compared, dissipative particle dynamics (DPD) and coupled lattice-Boltzmann/molecular dynamics (LB/MD). A general mapping scheme to match DPD to LB/MD is developed. For the weak coupling regime, analytic expressions for the flow profiles in the presence of partial-slip as well as no-slip boundary conditions are derived from the Poisson-Boltzmann and Stokes equations, which are in good agreement with the numerical results. The influence of electrofriction and partial slip on the flow profiles is discussed. PMID- 19566170 TI - Efficient solution of the self-consistent field theory for block copolymer fluids displaying Schulz-Flory polydispersity. AB - We develop a new formulation of polymer self-consistent field theory for polydisperse copolymer fluids satisfying the Schulz-Flory distribution. The result we obtain is remarkably simple and quickly solved numerically. The computational effort scales inversely with the width of the Schulz-Flory distribution. This suggests a significant saving in computational effort when compared with existing approximate solutions of the polymer self-consistent field equations for polydispersed systems. PMID- 19566171 TI - Silicon carbide nanostructures: a tight binding approach. AB - A tight-binding model Hamiltonian is newly parametrized for silicon carbide based on fits to a database of energy points calculated within the density functional theory approach of the electronic energy surfaces of nanoclusters and the total energy of bulk 3C and 2H polytypes at different densities. This TB model includes s and p angular momentum symmetries with nonorthogonal atomic basis functions. With the aid of the new TB model, minima of silicon carbide cagelike clusters, nanotubes, ring-shaped ribbons, and nanowires are predicted. Energetics, structure, growth sequences, and stability patterns are reported for the nanoclusters and nanotubes. The band structure of SiC nanotubes and nanowires indicates that the band gap of the nanotubes ranges from 0.57 to 2.38 eV depending on the chirality, demonstrating that these nanotubes are semiconductors or insulators. One type of nanowire is metallic, another type is semiconductor, and the rest are insulators. PMID- 19566172 TI - Self-diffusion and macroscopic diffusion of hydrogen in amorphous metals from first-principles calculations. AB - Diffusion of interstitial hydrogen plays a key role in potential uses for amorphous metals as membranes for hydrogen purification. We show how first principles-based methods can be used to characterize diffusion of interstitial H in amorphous metals using amorphous Fe(3)B as an example. Net transport of interstitial H is governed by the transport diffusion coefficient that appears in Fick's law. This diffusion coefficient is strongly dependent on the interstitial concentration, and is not equal to the self-diffusion coefficient except at dilute interstitial concentrations. Under conditions of practical interest, the concentrations of interstitial H in amorphous metals are nondilute so methods to determine the transport diffusion coefficient must be used if net mass transport is to be described. We show how kinetic Monte Carlo simulations of interstitial H diffusion that use rates derived from first-principles calculations can be used to assess both self- and transport diffusion coefficients of H in amorphous metals. These methods will be helpful in efforts to screen amorphous metal alloys as potential membranes for hydrogen purification. PMID- 19566173 TI - Adsorption and diffusion of Au atoms on the (001) surface of Ti, Zr, Hf, V, Nb, Ta, and Mo carbides. AB - The adsorption of atomic Au on the (001) surface of TiC, ZrC, HfC, VC, NbC, TaC, and delta-MoC and the mechanism of diffusion of this adatom through the surface have been studied in terms of a periodic density functional theory based approach. In all the cases, the Au adsorption energies are in the range of 1.90 2.35 eV. The moderately large adsorption energies allow the Au diffusion before desorption could take place. For TiC(001), ZrC(001), and HfC(001), atomic Au is adsorbed directly on top of C atoms and diffusion takes place along the diagonal of the squares formed by M-C-M-C atoms with the transition state located above the hollow sites. For the rest of transition metal carbides the situation is less simple with the appearance of more than one stable adsorption site, as for NbC and TaC, of a small energy barrier for diffusion around the most stable adsorption site and of a more complex diffusion pathway. The small energy barrier for diffusion around the most stable site will result in a highly mobile Au species which could be observed in scanning tunnel microscope experiments. After depositing Au on metal-carbide surfaces, there is a noticeable charge transfer from the substrate to the adsorbed Au atom. The electronic perturbations on Au increase when going from TiC to ZrC or TaC. Our results indicate that metal carbides should be better supports for the chemical activation of Au than metal oxides. PMID- 19566174 TI - Matrix effects on secondary ion emission from a room-temperature ionic liquid, 1 ethyl-3-methylimidazolium bis[trifluoromethanesulfonyl]imide. AB - The ionization mechanism of room-temperature ionic liquids has been investigated using time-of-flight secondary ion mass spectrometry in the temperature range of 15-300 K. Analyses of 1-ethyl-3-methylimidazolium bis[trifluoromethanesulfonyl]imide ([emim][Tf(2)N]) deposited on a Ni(111) substrate revealed that the [emim](+) and [Tf(2)N](-) yields increase together with the Ni(+) yield at monolayer coverage; no such increase was observed for the films deposited on a D(2)O spacer layer. Results indicated that the [emim][Tf(2)N] molecule is not perfectly ionized; the Ni(111) surface accepts (for [emim](+)) or donates (for [Tf(2)N](-)) an electron with higher efficiency than the counterion because of the metal band effect. This phenomenon might be induced by electrostatic interactions between the separated cation and anion during sputtering. It is also suggested that the sputtered Ni atom can be ionized nonadiabatically by the formation of a quasimolecule with adspecies. The multilayer of [emim][Tf(2)N] deposited at 15 K has a porous structure, resembling that of polar molecules, because of nonionic intermolecular interactions. The phase transition is identifiable, together with the morphological change in the crystalline film, from temperature evolutions of the secondary ion yields. PMID- 19566175 TI - Structure and interaction of flexible dendrimers in concentrated solution. AB - We study the influence of mutual interaction on the conformation of flexible poly(propyleneamine) dendrimers of fourth generation in concentrated solution. Mixtures of dendrimers with protonated and deuterated end groups are investigated by small-angle neutron scattering up to volume fractions of 0.23. This value is in the range of the overlap concentration of the dendrimers. The contrast between the solute and the solvent was varied by using mixtures of protonated and deuterated solvents. This allows us to investigate the partial structure factors of the deuterated dendrimers in detail. An analysis of the measured scattering intensities reveals that the shape of the flexible dendrimers is practically independent of the concentration in contrast to the pronounced conformational changes in flexible linear polymers. PMID- 19566176 TI - Theoretical study of solvent effects on the coil-globule transition. AB - The coil-globule transition of a polymer in a solvent has been studied using Monte Carlo simulations of a single chain subject to intramolecular interactions as well as a solvent-mediated effective potential. This solvation potential was calculated using several different theoretical approaches for two simple polymer/solvent models, each employing hard-sphere chains and hard-sphere solvent particles as well as attractive square-well potentials between some interaction sites. For each model, collapse is driven by variation in a parameter which changes the energy mismatch between monomers and solvent particles. The solvation potentials were calculated using two fundamentally different methodologies, each designed to predict the conformational behavior of polymers in solution: (1) the polymer reference interaction site model (PRISM) theory and (2) a many-body solvation potential (MBSP) based on scaled particle theory introduced by Grayce [J. Chem. Phys. 106, 5171 (1997)]. For the PRISM calculations, two well-studied solvation monomer-monomer pair potentials were employed, each distinguished by the closure relation used in its derivation: (i) a hypernetted-chain (HNC)-type potential and (ii) a Percus-Yevick (PY)-type potential. The theoretical predictions were each compared to results obtained from explicit-solvent discontinuous molecular dynamics simulations on the same polymer/solvent model systems [J. Chem. Phys. 125, 194904 (2006)]. In each case, the variation in the coil-globule transition properties with solvent density is mostly qualitatively correct, though the quantitative agreement between the theory and prediction is typically poor. The HNC-type potential yields results that are more qualitatively consistent with simulation. The conformational behavior of the polymer upon collapse predicted by the MBSP approach is quantitatively correct for low and moderate solvent densities but is increasingly less accurate for higher densities. At high solvent densities, the PRISM-HNC and MBSP approaches tend to overestimate, while the PRISM-PY approach underestimates the tendency of the solvent to drive polymer collapse. PMID- 19566177 TI - Entanglement network in nanoparticle reinforced polymers. AB - Polymer nanocomposites have been widely studied in efforts to engineer materials with mechanical properties superior to those of the pure polymer, but the molecular origins of the sought-after improved properties have remained elusive. An ideal polymer nanocomposite model has been conceived in which the nanoparticles are dispersed throughout the polymeric matrix. A detailed examination of topological constraints (or entanglements) in a nanocomposite glass provides new insights into the molecular origin of the improved properties in polymer nanocomposites by revealing that the nanoparticles impart significant enhancements to the entanglement network. Nanoparticles are found to serve as entanglement attractors, particularly at large deformations, altering the topological constraint network that arises in the composite material. PMID- 19566178 TI - Influences of streaming potential on cross stream migration of flexible polymer molecules in nanochannel flows. AB - We execute augmented Brownian dynamics (BD) simulation studies to show that the migration of flexible polyelectrolyte chains through nanochannels may be strongly governed by a complicated interplay between the electroviscous effects, near-wall interaction mechanisms, and diffusophoretic transport due to thermal gradients prevailing in the system. We further illustrate that in presence of mutually opposing pressure-driven and electro-osmotic transport and with an optimal choice of the ratio of the strength of these two flow fields, the electroviscous effects may turn out to be immensely consequential in strengthening the effective confinement of the polyelectrolyte. This, in turn may permit in achieving important biophysical feats that are otherwise obtainable only through significantly reduced nanochannel dimensions. PMID- 19566179 TI - Microcanonical analysis of adsorption of homopolymer chain on a surface. AB - The adsorption process of a homopolymer chain nongrafted on an attractive surface is numerically investigated using replica-exchange multicanonical Monte Carlo simulation. Based on the microcanonical analysis, the microcanonical entropy in the adsorption transition shows convex features. Correspondingly, with the coexistence of two phases, negative specific heat is also observed in the region, implying first-orderlike transition. The origin of the negative specific heat is due to the nonextensitivity of the energy in the system. This adsorption process has some similarities to the nucleation and growth mechanism in the crystalline process. Further study reveals that the transition type, either first- or second orderlike, during chain adsorption is strongly dependent on the chain length, interactions among segments, whether chain grafted on the surface, and force upon segments imposed by surface. PMID- 19566180 TI - The influence of shape on the glassy dynamics of hard nonspherical particle fluids. I. Dynamic crossover and elasticity. AB - We extend and apply the center-of-mass version of the microscopic naive mode coupling theory to study the ideal kinetic glass transition of dense fluids and suspensions composed of broad families of one-, two-, and three-dimensional hard nonspherical particles. A kinetic arrest diagram is constructed which indicates a dynamical crossover or onset of activated barrier hopping controlled transport. We find (quasi-) one-dimensional rods and rings form ideal glasses at the lowest volume fractions which decrease strongly with aspect ratio. Two-dimensional disks form ideal glasses at intermediate volume fractions which decrease slowly with the number of particles comprising the planar objects. Compact three-dimensional cluster particles exhibit a subtle nonmonotonic variation of the onset volume fraction that depends on their detailed shape, surface corrugation, and intraparticle interstitial volume. A strong correlation between the ideal kinetic arrest volume fraction and dimensionless compressibility (amplitude of density fluctuations) is predicted. The elastic shear modulus (transient localization length) grows (decreases) exponentially with volume fraction in a manner that becomes stronger as particle dimensionality increases. PMID- 19566181 TI - The influence of shape on the glassy dynamics of hard nonspherical particle fluids. II. Barriers, relaxation, fragility, kinetic vitrification, and universality. AB - We extend and apply the nonlinear Langevin equation theory of activated barrier hopping dynamics in glassy fluids and colloidal suspensions to study broad families of one-, two-, and three-dimensional hard nonspherical particles. Beyond the ideal kinetic arrest volume fraction, entropic barriers emerge with heights (alpha relaxation times, inverse diffusion constants) that increase nonlinearly (nonexponentially) with volume fraction and in a manner that becomes stronger with particle dimensionality. Partial collapse of the volume fraction dependence of barrier heights and reduced relaxation times of different particle shapes within a fixed dimensionality class are achieved based on a difference volume fraction variable that quantifies the distance from the ideal mode coupling theory dynamic crossover. However, the barrier, alpha relaxation time, and self diffusion constant results of all shapes are remarkably well collapsed onto a single universal master curve based on a theoretically motivated coupling constant which quantifies the renormalized mean square force on a tagged particle. The latter is determined mainly by the square of the intermolecular site-site contact value of the pair correlation function, thereby providing an explicit microscopic connection between local packing, binary collisions, and slow dynamics. A large variation of the dynamic fragility with particle shape is found with compact cluster particles being the most fragile. A kinetic glass transition map is constructed that is relevant to vitrification of laboratory colloidal suspensions. The possible relevance of the hard particle results for understanding the dynamic fragility of thermal van der Waals liquids is discussed. PMID- 19566182 TI - The interactions between surfactants and vesicles: dissipative particle dynamics. AB - The interactions between surfactants and vesicles formed by double-tail amphiphiles are investigated by the dissipative particle dynamics. As the surfactant concentration is increased, vesicle solubilization can be generally described by the three-stage hypothesis including vesicular region, vesicle micelle coexistence, and mixed micellar region. We study the partition of surfactants between the bilayer phase and the aqueous phase where a higher value of K indicates that more surfactant molecules are incorporated in the bilayer. It is found that ln(K(-1)) is proportional to the hydrophile-lipophile balance (HLB), which depicts the degree of hydrophilicity associated with a surfactant. As the overall hydrophilicity of surfactants increases, i.e., higher HLB, K declines and vice versa. When the amounts of surfactants reach a critical point, the solubilization begins and the coexistence of vesicles and mixed micelles is observed. Further increase in the surfactant concentration results in total collapse of the vesicle. Consistent with experimental observations, the three stages are identified through the vesicle size-surfactant concentration relation. Our simulations clearly demonstrate the process of the vesicle solubilization and confirm the validity of the three-stage hypothesis. PMID- 19566183 TI - Positive feedback regulation results in spatial clustering and fast spreading of active signaling molecules on a cell membrane. AB - Positive feedback regulation is ubiquitous in cell signaling networks, often leading to binary outcomes in response to graded stimuli. However, the role of such feedbacks in clustering, and in spatial spreading of activated molecules, has come to be appreciated only recently. We focus on the latter, using a simple model developed in the context of Ras activation with competing negative and positive feedback mechanisms. We find that positive feedback, in the presence of slow diffusion, results in clustering of activated molecules on the plasma membrane, and rapid spatial spreading as the front of the cluster propagates with a constant velocity (dependent on the feedback strength). The advancing fronts of the clusters of the activated species are rough, with scaling consistent with the Kardar-Parisi-Zhang equation in one dimension. Our minimal model is general enough to describe signal transduction in a wide variety of biological networks where activity in the membrane-proximal region is subject to feedback regulation. PMID- 19566184 TI - Rigidity, conformation, and solvation of native and oxidized tannin macromolecules in water-ethanol solution. AB - We studied by light scattering and small angle x-rays scattering (SAXS) conformations and solvation of plant tannins (oligomers and polymers) in mixed water-ethanol solutions. Their structures are not simple linear chains but contain about 6% of branching. Ab initio reconstruction reveals that monomers within a branch are closely bound pairwise. The chains are rather rigid, with the Kuhn length b = 13+/-3 nm, corresponding to about 35 linearly bound monomers. Contribution of solvation layer to SAXS intensity varies in a nonmonotonic way with ethanol content phi(A), which is an indication of amphipathic nature of tannin molecules. Best solvent composition phi(A)(B) is a decreasing function of polymerization degree N, in agreement with increasing water solubility of tannins with N. Polymers longer than b present a power-law behavior I approximately Q(-d) in the SAXS profile at high momentum transfer Q. The monotonic decrease in d with increasing phi(A) (from 2.4 in water to 1.9 in ethanol) points that the tannins are more compact in water than in ethanol, presumably due to attractive intramolecular interactions in water. Tannins were then oxidized in controlled conditions similar to real biological or food systems. Oxidation does not produce any intermolecular condensation, but generates additional intramolecular links. Some oxidation products are insoluble in water rich solvent. For that reason, we identify these species as a fraction of natural tannins called "T1" in the notation of Zanchi et al. [Langmuir 23, 9949 (2007)]. Within the fraction left soluble after oxidation, conformations of polymeric tannins, despite their higher rigidity, remain sensitive to solvent composition. PMID- 19566185 TI - Escape from cavity through narrow tunnel. AB - The paper deals with a diffusing particle that escapes from a cavity to the outer world through a narrow cylindrical tunnel. We derive expressions for the Laplace transforms of the particle survival probability, its lifetime probability density, and the mean lifetime. These results show how the quantities of interest depend on the geometric parameters (the cavity volume and the tunnel length and radius) and the particle diffusion coefficients in the cavity and in the tunnel. Earlier suggested expressions for the mean lifetime, which correspond to different escape scenarios, are contained in our result as special cases. In contrast to these expressions, our formula predicts correct asymptotic behavior of the mean lifetime in the absence of the cavity or tunnel. To test the accuracy of our approximate theory we compare the mean lifetime, the lifetime probability density, and the survival probability (the latter two are obtained by inverting their Laplace transforms numerically) with corresponding quantities found by solving numerically the three-dimensional diffusion equation, assuming that the cavity is a sphere and that the particle has the same diffusion coefficient in the cavity and in the tunnel. Comparison shows excellent agreement between the analytical and numerical results over a broad range of the geometric parameters of the problem. PMID- 19566186 TI - Comment on "On the Crooks fluctuation theorem and the Jarzynski equality" [J. Chem. Phys. 129, 091101 (2008)]. AB - It has recently been argued that a self-consistency condition involving the Jarzynski equality (JE) and the Crooks fluctuation theorem (CFT) is violated for a simple Brownian process [L. Y. Chen, J. Chem. Phys.129, 091101 (2008)]. This note adopts the definitions in the original formulation of the JE and CFT and demonstrates the contrary. PMID- 19566187 TI - Comment on "Dynamic aspects of the liquid-liquid phase transformation in silicon" [J. Chem. Phys. 129, 104503 (2008)]. AB - Recently, Jakse and Pasturel [J. Chem. Phys.129, 104503 (2008)] using first principles molecular dynamics simulations reported evidence for a fragile-to strong transition in supercooled liquid Si. This conclusion was based on the presence (absence) of a Boson peak in the low-density, strong liquid (high density, fragile liquid) phase. Based on existing experimental data it is pointed out that (i) the assignment of the Boson peak in the above reference is erroneous and (ii) the Boson peak strength is not generally useful for determining the strong or fragile character of a supercooled liquid. Therefore, the inference that liquid-liquid phase transition in supercooled liquid Si is a fragile-to strong transition is unjustifiable. PMID- 19566190 TI - High spatial resolution confocal microscope with independent excitation and detection scanning capabilities. AB - We present the design of a confocal microscope adapted for optical spectroscopy and imaging at cryogenic temperatures. This system is based on the existing approach of partly inserting the optical components of the microscope inside a helium-bath cryostat. It provides a spatial resolution approaching the diffraction limit with a mechanical stability allowing uninterrupted integration times exceeding 10 h and allows keeping track of a single emitter for unlimited periods of time. Furthermore, our design allows scanning the excitation spot and detection area independently of the sample position. This feature provides the means to perform probeless transport experiments on one-dimensional nanostructures. The scanning capabilities of this microscope are fully detailed and characterized using the photoluminescence of single nitrogen dyads at 4.5 K. PMID- 19566191 TI - A methodology for improving laser beam induced current images of dye sensitized solar cells. AB - Using the laser beam induced current (LBIC) technique for the study of solar cells and photovoltaic devices, it is possible to obtain images representing the different degrees of quantum efficiency observed on the surface of these elements. Dye sensitized solar cells (DSSCs) or photoelectrochemical solar cells, in contrast to those based on solid-solid interfaces, show a slow response to irradiance variations--up to tens of seconds. This is basically due to both viscous matter transport processes and load transfer. This response is inappreciable when the device is functioning continuously but when a LBIC scan is performed, in which the laser moves quickly from one point to another, the slow response produces a memory effect and the signal generated at one given point depends on the conversion efficiency coefficients of the previously excited positions, resulting in diffuse images and a lack of sharpness. This work presents a methodology to correct high-resolution LBIC mappings of DSSCs using an algorithm based on the kinetics of the discharge process of the irradiated zone. The validity of the proposed method has been evaluated by carrying out experiments where the algorithm has been applied to LBIC mappings. PMID- 19566192 TI - High-resolution, high-transmission soft x-ray spectrometer for the study of biological samples. AB - We present a variable line-space grating spectrometer for soft x-rays that covers the photon energy range between 130 and 650 eV. The optical design is based on the Hettrick-Underwood principle and tailored to synchrotron-based studies of radiation-sensitive biological samples. The spectrometer is able to record the entire spectral range in one shot, i.e., without any mechanical motion, at a resolving power of 1200 or better. Despite its slitless design, such a resolving power can be achieved for a source spot as large as (30 x 3000) microm2, which is important for keeping beam damage effects in radiation-sensitive samples low. The high spectrometer efficiency allows recording of comprehensive two-dimensional resonant inelastic soft x-ray scattering (RIXS) maps with good statistics within several minutes. This is exemplarily demonstrated for a RIXS map of highly oriented pyrolytic graphite, which was taken within 10 min. PMID- 19566193 TI - NIF unconverted light and its influence on DANTE measurements. AB - NIF laser facility produces 1053 nm light and a fundamental requirement for NIF is to give up to 1.8 MJ of 351 nm light for target physics experiments. The 351 nm light is provided by frequency tripling the 1053 nm light in nonlinear crystals in the final optics assembly, just before the laser light enters the target chamber. Since this tripling process is not 100% efficient, unconverted light from the conversion process also enters the chamber. This unconverted light does not directly hit the target but it can strike target support structures at average intensities of few TW/cm2 where it can generate unwanted, background soft x-rays that are measured by the soft x-ray diagnostic DANTE installed on the NIF target chamber. This diagnostic quantifies the x-radiation intensity inside the hohlraum by measuring the x-ray flux coming from the target's laser entrance hole. Due to its centimeter wide field of view, it integrates x-ray emission from both the flux exiting a hohlraum laser entrance hole and from the target support structure irradiated by residual 1omega and 2omega unconverted light. This work gives quantitative evaluations of the unconverted light for the first time and the effects on DANTE measurements for the future NIF tuning experiment called "Shock timing." Emission spectra are significantly modified leading to an overestimation of radiative temperature during the foot of the laser pulse since background x-rays are predominant in first two DANTE channel measurements. Mitigations of these effects by coating silicon paddle with plastic, using a smaller collimator to reduce DANTE field of view or eliminating DANTE channels in the analysis have been investigated. PMID- 19566194 TI - Near-infrared photon time-of-flight spectroscopy of turbid materials up to 1400 nm. AB - Photon time-of-flight spectroscopy (PTOFS) is a powerful tool for analysis of turbid materials. We have constructed a time-of-flight spectrometer based on a supercontinuum fiber laser, acousto-optical tunable filtering, and an InP/InGaAsP microchannel plate photomultiplier tube. The system is capable of performing PTOFS up to 1400 nm, and thus covers an important region for vibrational spectroscopy of solid samples. The development significantly increases the applicability of PTOFS for analysis of chemical content and physical properties of turbid media. The great value of the proposed approach is illustrated by revealing the distinct absorption features of turbid epoxy resin. Promising future applications of the approach are discussed, including quantitative assessment of pharmaceuticals, powder analysis, and calibration-free near infrared spectroscopy. PMID- 19566195 TI - Investigation of spectra unfolded for a filtered x-ray diode array with improved smoothness. AB - An unfolding algorithm using parabolic B-splines to smoothly reconstruct the soft x-ray spectra from the measurements of a filtered x-ray diode array is proposed. This array has been fabricated for the study of the soft x ray emitted by Z-pinch plasma. Unfolding results show that for the simulated noise-free blackbody spectra with temperature ranging from 20 to 250 eV, both the spectra and the total power are accurately recovered. Typical experimental waveforms along with the unfolded spectra and total power of x rays are presented. Possible defects due to the adoption of parabolic B-splines instead of conventionally used histograms are discussed. PMID- 19566196 TI - Three-axis rapid steering of optically propelled micro/nanoparticles. AB - This paper presents the design and implementation of a three-axis steering system, wherein a micro/nanoparticle is optically trapped and propelled to serve as a measurement probe. The actuators in the system consist of a deformable mirror enabling axial steering and a two-axis acousto-optic deflector for lateral steering. The actuation range is designed and calibrated to be over 20 microm along the two lateral axes and over 10 microm along the axial direction. The actuation bandwidth of the two lateral axes is over 50 kHz and the associated resolution is 0.016 nm (1sigma). The axial resolution is 0.16 nm, while the bandwidth is enhanced to over 3 kHz by model cancellation method. The performance of the three-axis steering system is illustrated by three sets of experiments. First, active Brownian motion control of the trapped probe is utilized to enhance trapping stability. Second, a large range three-dimensional (3D) steering of a 1.87 microm probe, contouring a complex 3D trajectory in a 6 x 6 x 4 microm3 volume, is demonstrated. Third, a closed-loop steering is implemented to achieve improved precision. PMID- 19566198 TI - Electron spectroscopy using two-dimensional electron detection and a camera in a single electron counting mode. AB - A brief description is given of an economical implementation of the read out of a two-dimensional detector in an electron spectrometer by a charge coupled device camera, using a pulse counting mode. Count rates up to 10 kHz can be handled in this way. A comparison with results obtained using a resistive anode detector is given for the case of electron scattering from Xe atoms. Good agreement was obtained between both detection techniques, establishing the validity of the method described here. PMID- 19566197 TI - A compact electron beam ion source with integrated Wien filter providing mass and charge state separated beams of highly charged ions. AB - A Wien filter was designed for and tested with a room temperature electron beam ion source (EBIS). Xenon charge state spectra up to the charge state Xe46+ were resolved as well as the isotopes of krypton using apertures of different sizes. The complete setup consisting of an EBIS and a Wien filter has a length of less than 1 m substituting a complete classical beamline setup. The Wien filter is equipped with removable permanent magnets. Hence total beam current measurements are possible via simple removal of the permanent magnets. In dependence on the needs of resolution a weak (0.2 T) or a strong (0.5 T) magnets setup can be used. In this paper the principle of operation and the design of the Wien filter meeting the requirements of an EBIS are briefly discussed. The first ion beam extraction and separation experiments with a Dresden EBIS are presented. PMID- 19566199 TI - A time-gating scintillation detector for the measurement of laser-induced fast neutrons. AB - A time-gating scintillation detector, in which a fast high voltage switch is used for gating a channel photomultiplier, was developed for a measurement of laser induced fast neutrons. The x rays generated from the intense femtosecond laser and the solid target interactions were suppressed selectively and a time-of flight signal of a laser-generated fast neutron was measured effectively. The detector was used successfully to measure the neutron yield of a femtosecond, deuterated, polystyrene plasma. PMID- 19566200 TI - Production and ion-ion cooling of highly charged ions in electron string ion source. AB - The scheme of an internal injection of Au atoms into the working space of the "Krion-2" electron string ion source (ESIS) was applied and tested. In this scheme Au atoms are evaporated from the thin tungsten wire surface in vicinity of the source electron string. Ion beams with charge states up to Au51+ were produced. Ion-ion cooling with use of C and O coolant ions was studied. It allowed increasing of the Au51+ ion yield by a factor of 2. Ions of Kr up to charge state 28+ were also produced in the source. Electron strings were first formed with injection electron energy up to 6 keV. Methods to increase the ESIS ion output are discussed. PMID- 19566201 TI - Digital ratemeter with fast response. AB - Digital ratemeter that operates according to a self-adjusting algorithm, which at low rates automatically switches from a preset count to the preset time method, is proposed. It combines the good properties of both methods concerning the accuracy of the measured average counting rate and the response time to sudden changes of rate. PMID- 19566202 TI - Method for analyzing E x B probe spectra from Hall thruster plumes. AB - Various methods for accurately determining ion species' current fractions using E x B probes in Hall thruster plumes are investigated. The effects of peak broadening and charge exchange on the calculated values of current fractions are quantified in order to determine the importance of accounting for them in the analysis. It is shown that both peak broadening and charge exchange have a significant effect on the calculated current fractions over a variety of operating conditions, especially at operating pressures exceeding 10(-5) torr. However, these effects can be accounted for using a simple approximation for the velocity distribution function and a one-dimensional charge exchange correction model. In order to keep plume attenuation from charge exchange below 30%, it is recommended that pz < or = 2, where p is the measured facility pressure in units of 10(-5) torr and z is the distance from the thruster exit plane to the probe inlet in meters. The spatial variation of the current fractions in the plume of a Hall thruster and the error induced from taking a single-point measurement are also briefly discussed. PMID- 19566203 TI - Generation of Mie size microdroplet aerosols with applications in laser-driven fusion experiments. AB - We have developed a tunable source of Mie scale microdroplet aerosols that can be used for the generation of energetic ions. To demonstrate this potential, a terawatt Ti:Al2O3 laser focused to 2 x 10(19) W/cm2 was used to irradiate heavy water (D2O) aerosols composed of micron-scale droplets. Energetic deuterium ions, which were generated in the laser-droplet interaction, produced deuterium deuterium fusion with approximately 2 x 10(3) fusion neutrons measured per joule of incident laser energy. PMID- 19566204 TI - State feedback control for adjusting the dynamic behavior of a piezoactuated bimorph atomic force microscopy probe. AB - We adjust the transient dynamics of a piezoactuated bimorph atomic force microscopy (AFM) probe using a state feedback controller. This approach enables us to adjust the quality factor and the resonance frequency of the probe simultaneously. First, we first investigate the effect of feedback gains on dynamic response of the probe and then show that the time constant of the probe can be reduced by reducing its quality factor and/or increasing its resonance frequency to reduce the scan error in tapping mode AFM. PMID- 19566205 TI - A passive long-wavelength infrared microscope with a highly sensitive phototransistor. AB - A passive scanning confocal microscope in the long-wavelength infrared (LWIR) region has been developed for sensitive imaging of spontaneous LWIR radiation by utilizing an ultrahighly sensitive detector, called the charge-sensitive infrared phototransistor (CSIP). The microscope consisted of room-temperature components including a Ge objective lens and liquid helium temperature components including a confocal pinhole, Ge relay lenses, and CSIP detector. With the microscope, thermal radiation (wavelength of 14.7 microm) spontaneously emitted by the object was studied with a spatial resolution of 25 microm. Clear passive LWIR imaging pictures were obtained by scanning a sample consisting of glass, Al foil, Ag paste, and Au. Clear passive LWIR image was also obtained even when the sample surface was covered by a GaAs or Si plate. This work suggests usefulness of CSIP detectors for application of passive LWIR microscopy. PMID- 19566206 TI - Concept for support and cleavage of brittle crystals. AB - We report on sample holders for crystals to be cleaved for the preparation of surfaces with large atomically flat terraces. The concept for mounting sample crystals is based on a vicelike clamping mechanism to securely hold the crystal in position while reducing the risk of fragmentation. Sample holders based on this concept and made of suitable materials allow preparation and cleavage of crystals in the ultrahigh vacuum at high or low temperatures. To cleave the crystal, we employ a scalpel blade mounted on a wobble stick to generate a highly localized stress field initiating the cleavage process. The sample holders are used for experiments of highest resolution scanning force microscopy, however, the concept can be transferred to any other system where cleavage faces of crystals are of interest. Exemplarily, scanning force microscopy results demonstrate that (111) cleavage faces of CaF2 crystals can be prepared with steps only a few F-Ca-F triple-layers high and atomically flat terraces extending over areas of several microm2. PMID- 19566207 TI - Ultrastable combined atomic force and total internal reflection fluorescence microscope [corrected]. AB - Combining atomic force microscope (AFM) with other microscopy techniques has expanded the range of potential applications for single molecule investigations dramatically. Particularly hybrid instruments with total internal reflection fluorescence (TIRF) excitation have opened new routes in life sciences. Here we present a novel design for such a hybrid microscope, which overcomes the limitations of conventional combinations caused by their limited mechanical stability. A thorough analysis of the noise spectra and a comparison of the different designs and the different operation modes are given. With this instrument we demonstrate single molecule manipulation by AFM and simultaneous TIRF imaging. PMID- 19566208 TI - Making a commercial atomic force microscope more accurate and faster using positive position feedback control. AB - This paper presents experimental implementation of a positive position feedback (PPF) control scheme for vibration and cross-coupling compensation of a piezoelectric tube scanner in a commercial atomic force microscope (AFM). The AFM is a device capable of generating images with extremely high resolutions down to the atomic level. It is also being used in applications that involve manipulation of matter at a nanoscale. Early AFMs were operated in open loop. Consequently, they were susceptible to piezoelectric creep, thermal drift, hysteresis nonlinearity, and scan-induced vibration. These effects tend to distort the generated image and slow down the scanning speed of the device. Recently, a new generation of AFMs has emerged that utilizes position sensors to measure displacements of the scanner in three dimensions. These AFMs are equipped with feedback control loops that work to minimize the adverse effects of hysteresis, piezoelectric creep, and thermal drift on the obtained image using proportional plus-integral (PI) controllers. These feedback controllers are often not designed to deal with the highly resonant nature of an AFM's scanner nor with the cross coupling between various axes. In this paper we illustrate the improvement in accuracy and imaging speed that can be achieved by using a properly designed feedback controller such as a PPF controller. Such controllers can be incorporated into most modern AFMs with minimal effort since they can be implemented in software with the existing hardware. Experimental results show that by implementing the PPF control scheme, relatively good images in comparison with a well-tuned PI controller can still be obtained up to line scan of 60 Hz. PMID- 19566209 TI - Thin film nanocalorimeter for heat capacity measurements of 30 nm films. AB - A silicon nitride membrane-based nanocalorimeter is described for measuring the heat capacity of 30 nm films from 300 mK to 800 K and in high magnetic fields with absolute accuracy approximately 2%. The addenda heat capacity of the nanocalorimeter is less than 2 x 10(-7) J/K at room temperature and 2 x 10(-10) J/K at 2.3 K. This is more than ten times smaller than any existing calorimeter suitable for measuring thin films over this wide temperature range. The heat capacities of thin Cu and Au films are reported and agree with bulk values. The thermal conductivity of the thin low stress silicon nitride is substantially smaller than thicker membranes while the specific heat is enhanced below 20 K. Design of the nanocalorimeter will be discussed along with fabrication details and calibration results. PMID- 19566210 TI - Diagnoses of coaxial probes in shock compression. AB - A miniature diagnostic apparatus, which consists of a target, a guiding tube, a nail-gun, and a simple base, has been developed in the proposed research to estimate batches of pinlike coaxial probes with selectable collision speeds, 198.3, 361.0, and 420.6 mps, corresponding to thrust capabilities of certain minibullets. This work aims at filling the gap between typical two stage light gas gun (>500 mps) and pendulum machine (<100 mps) with low cost and risk in realistic shock compressions. As a part of the experiment, the kappa coefficient and the statistical agreement and reliability are all evaluated. PMID- 19566211 TI - Cancellation of environmental effects in resonant mass sensors based on resonance mode and effective mass. AB - A novel technique is developed to cancel the effect of environmental parameters, e.g., temperature and humidity, in resonant mass sensing. Utilizing a single resonator, the environmental cancellation is achieved by monitoring a pair of resonant overtones and the effective sensed mass in those overtones. As an eminent advantage, especially compared to dual-mode temperature compensation techniques, the presented technique eliminates any need for previously measured look-up tables or fitting the measurement data. We show that a resonant cantilever beam is an appropriate platform for applying this technique, and derive an analytical expression to relate the actual and effective sensed masses on a cantilever beam. Thereby, it is shown that in applying the presented technique successfully, the effective sensed masses must not be the same in the investigated pair of resonance overtones. To prove the feasibility of the proposed technique, flexural resonance frequencies of a silicon cantilever are measured before and after loading with a strip of photoresist. Applying the presented technique shows significant reductions in influence of environmental parameters, with the temperature and humidity coefficients of frequency being improved from -19.5 to 0.2 ppm degrees C(-1) and from 0.7 to -0.03 ppm %RH(-1), respectively. PMID- 19566212 TI - Setup for optimized grazing incidence x-ray absorption experiments on thin films on substrates. AB - We present a state-of-the-art experimental apparatus and a proper setup to perform x-ray absorption spectroscopy (XAS) experiments in grazing incidence mode. This geometry is appropriate for doped thin films or interfaces buried at moderate depth in a thick matrix, whenever the scattering and/or fluorescence from the matrix has to be strongly attenuated. Both the calculation and the experimental data demonstrate that the specific setup that consists in a grazing incidence and grazing collection geometry is extremely advantageous. In fact, with respect to the standard geometry used to perform XAS experiments in fluorescence mode, the present setup allows an enhancement in the interesting fluorescence signal from the surface layer without a corresponding increase in the elastic scattering contribution from the matrix. The sample holder especially designed for this kind of experiment can work in vacuum and at low temperature. An easy and quick automatic sample alignment procedure is detailed. PMID- 19566213 TI - End-compensated magnetostatic cavity for polarized 3He neutron spin filters. AB - We have expanded upon the "Magic Box" concept, a coil driven magnetic parallel plate capacitor constructed out of mu-metal, by introducing compensation sections at the ends of the box that are tuned to limit end-effects similar to those of short solenoids. This ability has reduced the length of the magic box design without sacrificing any loss in field homogeneity, making the device far more applicable to the often space limited neutron beam line. The appeal of the design beyond affording longer polarized 3He lifetimes is that it provides a vertical guide field, which facilitates neutron spin transport for typical polarized beam experiments. We have constructed two end-compensated magic boxes of dimensions 28.4 x 40 x 15 cm3 (length x width x height) with measured, normalized volume averaged transverse field gradients ranging from 3.3 x 10(-4) to 6.3 x 10(-4) cm( 1) for cell sizes ranging from 8.1 x 6.0 to 12.0 x 7.9 cm2 (diameter x length), respectively. PMID- 19566214 TI - Sizing and burn time measurements of micron-sized metal powders. AB - Detailed ignition and combustion mechanisms are needed to develop optimized propellant and energetic formulations using micron-sized metal powders, such as aluminum. Combustion researchers have traditionally used relatively coarse metal particles to characterize the burn time dependence on particle size. However, measurements of burn times for particles below 10 microm in diameter are still needed for aluminum powders and other metal fuels. The apparatus described here sizes the particles just before the ignition event, providing a direct correlation between individual particle size and its burn time. Two lasers were utilized: a 785 nm laser diode for sizing the particles and a 125 W CO2 laser for particle ignition. The particles crossed the 785 nm laser beam just before crossing the CO2 laser beam. The particle size was determined from the amplitude of the scattered 785 nm light pulse. The burn time was determined from the duration of the visible light emission produced from the ignited particle. The in situ measured particle size distributions compared well with the size distributions measured for the same powders by a commercial instrument using low angle laser light scattering. Our measurements with two nominally spherical aluminum powders, suggest that the burn times increase from 0.5 to approximately 2.5 ms as the particle diameters increase from 3 to 8 microm. PMID- 19566215 TI - RF testbed for thermoacoustic tomography. AB - Thermoacoustic signal excitation is a function of intrinsic tissue properties and illuminating electric field. De-ionized (DI) water is a preferred acoustic coupling medium for thermoacoustics because acoustic and electromagnetic waves propagate in DI water with very little loss. We have designed a water-filled testbed propagating a controlled electric field with respect to pulse shape, power, and polarization. Directional coupler line sections permit measurement of incident, reflected, and transmitted powers. Both S-parameters and E(y) measurement show that the electric-field distribution is relatively uniform in testbed. Comparing baseline power measurements to those taken with a test object in place yields power loss in the object, which should correlate to thermoacoustic signal strength. Moreover, power loss--and therefore thermoacoustic computerized tomography signal strength--is sensitive to the orientation of the object to the polarization of the electric field. This testbed will enable quantitative characterization of the thermoacoustic contrast mechanism in ex vivo tissue specimens. PMID- 19566216 TI - The PLATO Dome A site-testing observatory: power generation and control systems. AB - The atmospheric conditions above Dome A, a currently unmanned location at the highest point on the Antarctic plateau, are uniquely suited to astronomy. For certain types of astronomy Dome A is likely to be the best location on the planet, and this has motivated the development of the Plateau Observatory (PLATO). PLATO was deployed to Dome A in early 2008. It houses a suite of purpose built site-testing instruments designed to quantify the benefits of Dome A site for astronomy, and science instruments designed to take advantage of the observing conditions. The PLATO power generation and control system is designed to provide continuous power and heat, and a high-reliability command and communications platform for these instruments. PLATO has run and collected data throughout the winter 2008 season completely unattended. Here we present a detailed description of the power generation, power control, thermal management, instrument interface, and communications systems for PLATO, and an overview of the system performance for 2008. PMID- 19566217 TI - Stable voltage source for Penning trap experiments. AB - A voltage reference has been developed to bias ring electrodes of two Penning traps between -90 and 0 V. For output voltages near -90 V, the Allan deviation of the system's voltage instability is less than 1 part in 10(8) over all time scales shorter than 10(4) s. For averaging times longer than several seconds, the system's stability is determined almost completely by the noise, drift, and aging of the zener diodes in the array of voltage reference integrated circuits. For shorter averaging times, active filters built into the new system significantly reduce the intrinsic noise of the zener diodes. The system makes it possible to continuously adjust the ring voltages for frequency locking the axial motion in the two Penning traps. By keeping electrical noise highly correlated between the two traps, measurement uncertainty should be reduced for precision experiments such as Penning trap mass spectrometry. PMID- 19566218 TI - Simultaneous measurements of the specific heat and thermal conductivity of suspended thin samples by transient electrothermal method. AB - The electrothermal technique is developed to simultaneously measure the specific heat and thermal conductivity of individual thin samples suspended across two heat sinks, resorting to pulsed direct currents with or without a dc offset. The temperature evolution due to Joule self-heating is recorded and compared with the numerical solutions of transient heat conduction equations using the finite volume method. The thermal conductivity is determined by the steady temperature level and the specific heat by the transient temperature rise or relaxation. This technique is applied to a 10 microm thick platinum wire and the thermal conductivity and specific heat are in good agreement with the literature values. In addition, the influences of thermal radiation and thermal boundary resistance between the sample and heat sinks on the experimental results are discussed. PMID- 19566219 TI - Heat transfer analysis for peripheral blood flow measurement system. AB - Some disorders such as circulatory disease and metabolic abnormality cause many problems to peripheral blood flow condition. Therefore, frequent measurement of the blood flow condition is bound to contribute to precaution against those disorders and to control of conditions of the diseases. We propose a convenient means of blood flow volume measurement at peripheral part, such as fingertips. Principle of this measurement is based on heat transfer characteristics of peripheral part containing the blood flow. Transition response analysis of skin surface temperature has provided measurement model of the peripheral blood flow volume. We developed the blood flow measurement system based on that model and evaluated it by using artificial finger under various temperature conditions of ambience and internal fluid. The evaluation results indicated that proposed method could estimate the volume of the fluid regardless of temperature condition of them. Finally we applied our system to real finger testing and have obtained results correlated well with laser Doppler blood flow meter values. PMID- 19566220 TI - Static and dynamic analysis of a four-tube piezoelectric actuator. AB - Piezoelectric tubes with film electrodes on their outer and inner surfaces can be used to compose multitube actuators. The actuator of four piezoelectric tubes can substitute the traditional single-tube actuator with quartered electrodes which has been widely used in scanning probe microscopy and piezoelectric motors. In this article, the static deflection, static bending moment, resonant frequency, and dynamic deflection of the four-tube actuator are all studied in detail. The comparison between this actuator and the traditional single-tube actuator is also done and the results show that the four-tube actuator has better performances under certain conditions. The experiment results of a prototype actuator testified the validation of the analysis. PMID- 19566221 TI - Quasistatic displacement self-sensing method for cantilevered piezoelectric actuators. AB - Piezoelectric meso- and microactuator systems required for manipulation or assembly of microscale objects demand reliable force and/or displacement information. Available sensors are prone to dimension restrictions or precision limitation. Self-sensing method, based on the electric charge measurement, may represent a solution in terms of cost-effectiveness and integration, the actuator performing simultaneously as its own sensor. This paper presents a self-sensing method dedicated to free uni- and bimorph piezocantilevers but can also be adapted to other piezoactuator types. The integrated electric current, used to convert the charge, can be compensated against piezoelectric material nonlinearities to provide accurate displacement information. The advantages relative to existing self-sensing methods consist in the ability to keep this displacement information for long-term periods (more than a thousand seconds) and in the reduction in signal noise. After introductive issues related to the method the base principle allowing the estimation of tip displacement is presented. Then, the identification procedure of the estimator parameters is depicted and representative experimental results are shown. Finally, a series of aspects related to electronic circuits are discussed, useful for successful system implementation. PMID- 19566222 TI - Experimental device to measure the electrical and optical properties of radiochromic films as a function of temperature. AB - In the last decade, radiochromic films (RCF) have been used extensively in medical physics for evaluating uniformity of radiation beams and dose distributions. These films are very sensitive to changes in temperature; therefore, one needs to know its thermal behavior to use them efficiently. Normally, the thermal assessment is done using optical methods on previously irradiated samples. Here, we report the use of a complementary dielectric method. We designed an experimental device that allows us to measure, simultaneously, the dielectric and optical properties as a function of temperature. We performed the measurements in real-time to temperature increases from 27 to 48 degrees C of EBT and MD-55 RCF previously exposed to UV. We found for both films a decrease in the real part of the dielectric permittivity as the temperature increases, but an increment and decrease in the dielectric loss factor for the EBT and MD-55 film, respectively. PMID- 19566224 TI - Research on three dimensional machining effects using atomic force microscope. AB - This research studies the use of scanning probe microscope as the tool to manufacture three dimensional nanoscale objects. We modified a commercial atomic force microscope (AFM) and replaced the original probe control system with a personal computer (PC) based controller. The modified system used the scanning probe in the AFM for the cutting tool and used the PC controller to control work piece. With the new controller, one could implement multiaxes motion control to perform trajectory planning and to test various cutting strategies. The experiments discovered that the debris can coalesce with the sample material and cause tremendous problem in the nanomachining process. This research thus proposed to make use of this material and developed a piling algorithm to not only cut but also pile up the debris in a favorable way for steric shaping. The experimental results showed that the proposed cutting and shaping algorithm can produce nano-objects as high as a few hundred nanometers. The probe tip typically wears down to around 500 microm diameter after the machining process, putting a limit on the machining resolution. The vertical resolution can achieve less than 10 nm without controlled environment. PMID- 19566223 TI - Development of a dual-modal tissue diagnostic system combining time-resolved fluorescence spectroscopy and ultrasonic backscatter microscopy. AB - We report a tissue diagnostic system which combines two complementary techniques of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) and ultrasonic backscatter microscopy (UBM). TR-LIFS evaluates the biochemical composition of tissue, while UBM provides tissue microanatomy and enables localization of the region of diagnostic interest. The TR-LIFS component consists of an optical fiber based time-domain apparatus including a spectrometer, gated multichannel plate photomultiplier, and fast digitizer. It records the fluorescence with high sensitivity (nM concentration range) and time resolution as low as 300 ps. The UBM system consists of a transducer, pulser, receiving circuit, and positioning stage. The transducer used here is 45 MHz, unfocused, with axial and lateral resolutions 38 and 200 microm. Validation of the hybrid system and ultrasonic and spectroscopic data coregistration were conducted both in vitro (tissue phantom) and ex vivo (atherosclerotic tissue specimens of human aorta). Standard histopathological analysis of tissue samples was used to validate the UBM-TRLIFS data. Current results have demonstrated that spatially correlated UBM and TR-LIFS data provide complementary characterization of both morphology (necrotic core and calcium deposits) and biochemistry (collagen, elastin, and lipid features) of the atherosclerotic plaques at the same location. Thus, a combination of fluorescence spectroscopy with ultrasound imaging would allow for better identification of features associated with tissue pathologies. Current design and performance of the hybrid system suggests potential applications in clinical diagnosis of atherosclerotic plaque. PMID- 19566225 TI - Development of novel hybrid flexure-based microgrippers for precision micro object manipulation. AB - This paper describes the process of developing a microgripper that is capable of high precision and fidelity manipulation of micro-objects. The design adopts the concept of flexure-based hinges on its joints to provide the rotational motion, thus eliminating the inherent nonlinearities associated with the application of conventional rigid hinges. A combination of two modeling techniques, namely, pseudorigid body model and finite element analysis was utilized to expedite the prototyping procedure, which leads to the establishment of a high performance mechanism. A new hybrid compliant structure integrating cantilever beam and flexural hinge configurations within microgripper mechanism mainframe has been developed. This concept provides a novel approach to harness the advantages within each individual configuration while mutually compensating the limitations inherent between them. A wire electrodischarge machining technique was utilized to fabricate the gripper out of high grade aluminum alloy (Al 7075T6). Experimental studies were conducted on the model to obtain various correlations governing the gripper performance as well as for model verification. The experimental results demonstrate high level of compliance in comparison to the computational results. A high amplification characteristic and maximum achievable stroke of 100 microm can be achieved. PMID- 19566226 TI - Accurate noncontact calibration of colloidal probe sensitivities in atomic force microscopy. AB - The absolute force sensitivities of colloidal probes comprised of atomic force microscope, or AFM, cantilevers with microspheres attached to their distal ends are measured. The force sensitivities are calibrated through reference to accurate electrostatic forces, the realizations of which are described in detail. Furthermore, the absolute accuracy of a common AFM force calibration scheme, known as the thermal noise method, is evaluated. It is demonstrated that the thermal noise method can be applied with great success to colloidal probe calibration in air and in liquid to yield force measurements with relative standard uncertainties below 5%. Techniques to combine the electrostatics-based determination of the AFM force sensitivity with measurements of the colloidal probe's thermal noise spectrum to compute noncontact estimates of the displacement sensitivity and spring constant are also developed. PMID- 19566227 TI - The bone diagnostic instrument III: testing mouse femora. AB - Here we describe modifications that allow the bone diagnostic instrument (BDI) [P. Hansma et al., Rev. Sci. Instrum. 79, 064303 (2008); Rev. Sci. Instrum. 77, 075105 (2006)], developed to test human bone, to test the femora of mice. These modifications include reducing the effective weight of the instrument on the bone, designing and fabricating new probe assemblies to minimize damage to the small bone, developing new testing protocols that involve smaller testing forces, and fabricating a jig for securing the smaller bones for testing. With these modifications, the BDI was used to test the hypothesis that short-term running has greater benefit on the mechanical properties of the femur for young growing mice compared to older, skeletally mature mice. We measured elastic modulus, hardness, and indentation distance increase (IDI), which had previously been shown to be the best discriminators in model systems known to exhibit differences in mechanical properties at the whole bone level. In the young exercised murine femora, the IDI was significantly lower than in young control femora. Since IDI has a relation to postyield properties, these results suggest that exercise during bone development increases post yield mechanical competence. We were also able to measure effects of aging on bone properties with the BDI. There was a significant increase in the IDI, and a significant decrease in the elastic modulus and hardness between the young and old groups. Thus, with the modifications described here, the BDI can take measurements on mouse bones and obtain statistically significant results. PMID- 19566229 TI - Corrected direct force balance method for atomic force microscopy lateral force calibration. AB - This paper reports corrections and improvements of the previously reported direct force balance method (DFBM) developed for lateral calibration of atomic force microscopy. The DFBM method employs the lateral force signal obtained during a force-distance measurement on a sloped surface and relates this signal to the applied load and the slope of the surface to determine the lateral calibration factor. In the original publication [Rev. Sci. Instrum. 77, 043903 (2006)], the tip-substrate contact was assumed to be pinned at the point of contact, i.e., no slip along the slope. In control experiments, the tip was found to slide along the slope during force-distance curve measurement. This paper presents the correct force balance for lateral force calibration. PMID- 19566228 TI - A direct micropipette-based calibration method for atomic force microscope cantilevers. AB - In this report, we describe a direct method for calibrating atomic force microscope (AFM) cantilevers with the micropipette aspiration technique (MAT). A closely fitting polystyrene bead inside a micropipette is driven by precisely controlled hydrostatic pressures to apply known loads on the sharp tip of AFM cantilevers, thus providing a calibration at the most functionally relevant position. The new method is capable of calibrating cantilevers with spring constants ranging from 0.01 to hundreds of newtons per meter. Under appropriate loading conditions, this new method yields measurement accuracy and precision both within 10%, with higher performance for softer cantilevers. Furthermore, this method may greatly enhance the accuracy and precision of calibration for colloidal probes. PMID- 19566230 TI - Diffractive grating structure for coherent light source production. AB - A diffractive structure of cavities is designed to produce resonances at very high frequencies. This type of cavity structure may be useful for producing controllable coherent synchrotron radiation by microbunching the particle beam. PMID- 19566231 TI - A rapid method for flow-ready cylindrical microchannel fabrication. AB - Most current methods of microchannel fabrication are expensive and time consuming or result in noncylindrical channels, which is undesirable for many applications. A rapid inexpensive method for the fabrication of flow-ready cylindrical polymer microchannels from polydimethylsiloxane (PDMS) is presented here. In this method, cylindrical needles are used as molds for the channels and also function as flow delivery devices after fabrication. Validation of channel function and smoothness can be accomplished by comparing experimental data to theoretical models. One model was previously developed by the author to predict the incipient motion of a glass sphere in contact with a perfectly smooth PDMS surface and deviations from the model are expected to be a function of channel roughness. The data collected in the present channels show fairly good agreement with the theoretical model, indicating a relatively smooth and consistent surface. In addition, detailed SEM images of the channel showed that the internal surface was qualitatively very smooth. PMID- 19566232 TI - Proximity detector circuits: an alternative to tunnel diode oscillators for contactless measurements in pulsed magnetic field environments. AB - A radio frequency oscillator circuit based on a proximity detector integrated circuit is described as an alternative for the traditional tunnel diode oscillator used for pulsed magnetic field measurements at low temperatures. The proximity detector circuit design, although less sensitive than tunnel diode oscillator circuits, has a number of essential advantages for measurements in the extreme environments of pulsed magnetic fields. These include the insensitivity of operation to voltages induced in the inductor coil, the elimination of a diode bias circuit and tuning, and a broad dynamic range of resonant frequency variation. The circuit has been successfully applied to measure the superconducting upper critical field in Ba(0.55)K(0.45)Fe2As2 single crystals up to 60 T. PMID- 19566236 TI - Empirical analysis of Internet telephone network: from user ID to phone. AB - In order to study the interaction between different communication networks, in this paper, personal computer (PC)-to-phone log data in the year of 2007 are collected from UUCALL database and described as an ID-to-phone bipartite network (ItPBN). The ItPBN contains one giant component (GC) and a large number of satellitic components (SCs), both of which are carefully analyzed. It is found that the ItPBN has power-law incoming/outgoing degree distributions as well as a power-law clustering function (by proposing a new definition of clustering coefficient) indicating a hierarchical and modular structure of the ItPBN. Furthermore, the fact that most of the weak links always surrounding those ID nodes of large degree in the GC suggests that weak links may be more important to keep the structure of the GC than those strong ones. More interestingly, it is also revealed that there is strong correlation between many statistical properties of different SCs and their size, these extra information may be very useful in modeling the ItPBN in the future. PMID- 19566237 TI - Spatial order prevails over memory in boosting cooperation in the iterated prisoner's dilemma. AB - This paper presents some results on a spatial version of the iterated prisoner's dilemma in which every player imitates in any iteration the optimal strategy of its neighbors. Neighbors are defined with different degrees of random variation (initial rewiring) based on a square lattice, and optimal is defined with different degrees of memory, ranging from only the single preceding iteration up to all preceding iterations. It is concluded that memory notably stimulates cooperation in the iterated prisoner's dilemma played in ordered lattices, but it is unable to boost cooperation as the wiring network becomes highly disordered. PMID- 19566239 TI - Recurrences determine the dynamics. AB - We show that under suitable assumptions, Poincare recurrences of a dynamical system determine its topology in phase space. Therefore, dynamical systems with the same recurrences are dynamically equivalent. This conclusion can be drawn from a theorem proved in this paper which states that the recurrence matrix determines the topology of closed sets. The theorem states that if a set of points M is mapped onto another set N, such that two points in N are closer than some prescribed fixed distance if and only if the corresponding points in M are closer than some, in general different, prescribed fixed distance, then both sets are homeomorphic, i.e., identical up to a continuous change in the coordinate system. The theorem justifies a range of methods in nonlinear dynamics which are based on recurrence properties. PMID- 19566238 TI - Frequently asked questions about global modeling. AB - When a global model is attempted from experimental data, some preprocessing might be required. Therefore it is only natural to wonder what kind of effects the preprocessing might have on the modeling procedure. This concern is manifested in the form of recurrent frequently asked questions, such as "how does the preprocessing affect the underlying dynamics?" This paper aims at providing answers to important questions related to (i) data interpolation, (ii) data smoothing, (iii) data-estimated derivatives, (iv) model structure selection, and (v) model validation. The answers provided will hopefully remove some of those doubts and one shall be more confident not only on global modeling but also on various data analyses which may be also dependent on data preprocessing. PMID- 19566240 TI - Anticipating synchronization of a class of chaotic systems. AB - This paper studies the anticipating synchronization of a class of coupled chaotic systems. The asymptotic stability and exponential stability criteria for the involved error dynamical system are established by means of model transformation incorporated with Lyapunov-Krasovskii functional and linear matrix inequality. Based on the proposed stability conditions the coupling strength is then explicitly designed in terms of system parameters and anticipating time. Numerical simulations are presented to verify the theoretical results. PMID- 19566241 TI - Cluster synchronization in community networks with nonidentical nodes. AB - In this paper dynamical networks with community structure and nonidentical nodes and with identical local dynamics for all individual nodes in each community are considered. The cluster synchronization of these networks with or without time delay is studied by using some feedback control schemes. Several sufficient conditions for achieving cluster synchronization are obtained analytically and are further verified numerically by some examples with chaotic or nonchaotic nodes. In addition, an essential relation between synchronization dynamics and local dynamics is found by detailed analysis of dynamical networks without delay through the stage detection of cluster synchronization. PMID- 19566242 TI - Inverse synchronizations in coupled time-delay systems with inhibitory coupling. AB - Transitions between inverse anticipatory, inverse complete, and inverse lag synchronizations are shown to occur as a function of the coupling delay in unidirectionally coupled time-delay systems with inhibitory coupling. We have also shown that the same general asymptotic stability condition obtained using the Krasovskii-Lyapunov functional theory can be valid for the cases where (i) both the coefficients of the Delta(t) (error variable) and Delta(tau)=Delta(t tau) (error variable with delay) terms in the error equation corresponding to the synchronization manifold are time independent and (ii) the coefficient of the Delta term is time independent, while that of the Delta(tau) term is time dependent. The existence of different kinds of synchronization is corroborated using similarity function, probability of synchronization, and also from changes in the spectrum of Lyapunov exponents of the coupled time-delay systems. PMID- 19566243 TI - Chaos synchronization basing on symbolic dynamics with nongenerating partition. AB - Using symbolic dynamics and information theory, we study the information transmission needed for synchronizing unidirectionally coupled oscillators. It is found that when sustaining chaos synchronization with nongenerating partition, the synchronization error will be larger than a critical value, although the required coupled channel capacity can be smaller than the case of using a generating partition. Then we show that no matter whether a generating or nongenerating partition is in use, a high-quality detector can guarantee the lead of the response oscillator, while the lag responding can make up the low precision of the detector. A practicable synchronization scheme basing on a nongenerating partition is also proposed in this paper. PMID- 19566244 TI - Parameter identification and projective synchronization between different chaotic systems. AB - In this paper, adaptive generalized projective synchronization and parameter identification in different chaotic systems are carefully revisited. We use a concrete counterexample to demonstrate that the result in a previous work [R. Li, W. Xu, and S. Li, Phys. Lett. A 367, 199 (2007)] is imperfect, where a scheme of generalized projective synchronization is proposed for parameter identification with some drawbacks on ignoring the conditions which ensure the parameter convergence. We further discuss the two conditions of parameter convergence, which are linear independence and persistent excitation. A special relationship between them is addressed to estimate unknown model parameters effectively. PMID- 19566245 TI - Influence of thermal effects on buoyancy-driven convection around autocatalytic chemical fronts propagating horizontally. AB - The spatiotemporal dynamics of vertical autocatalytic fronts traveling horizontally in thin solution layers closed to the air can be influenced by buoyancy-driven convection induced by density gradients across the front. We perform here a combined experimental and theoretical study of the competition between solutal and thermal effects on such convection. Experimentally, we focus on the antagonistic chlorite-tetrathionate reaction for which solutal and thermal contributions to the density jump across the front have opposite signs. We show that in isothermal conditions the heavier products sink below the lighter reactants, providing an asymptotic constant finger shape deformation of the front by convection. When thermal effects are present, the hotter products, on the contrary, climb above the reactants for strongly exothermic conditions. These various observations as well as the influence of the relative weight of the solutal and thermal effects and of the thickness of the solution layer on the dynamics are discussed in terms of a two-dimensional reaction-diffusion convection model parametrized by a solutal R(C) and a thermal R(T) Rayleigh number. PMID- 19566246 TI - Dynamics in scheduled networks. AB - When studying real or virtual systems through complex networks theories, usually time restrictions are neglected, and a static structure is defined to characterize which node is connected to which other. However, this approach is oversimplified, as real networks are indeed dynamically modified by external mechanisms. In order to bridge the gap, in this work we present a scheduled network formalism, which takes into account such dynamical modifications by including generic time restrictions in the structure of an extended adjacency matrix. We present some of its properties and apply this formalism to the specific case of the air transportation network in order to analyze its efficiency. Real data are used at this point. We finally discuss on the applicability of this formalism to other complex systems. PMID- 19566247 TI - Delay-induced multiple stochastic resonances on scale-free neuronal networks. AB - We study the effects of periodic subthreshold pacemaker activity and time-delayed coupling on stochastic resonance over scale-free neuronal networks. As the two extreme options, we introduce the pacemaker, respectively, to the neuron with the highest degree and to one of the neurons with the lowest degree within the network, but we also consider the case when all neurons are exposed to the periodic forcing. In the absence of delay, we show that an intermediate intensity of noise is able to optimally assist the pacemaker in imposing its rhythm on the whole ensemble, irrespective to its placing, thus providing evidences for stochastic resonance on the scale-free neuronal networks. Interestingly thereby, if the forcing in form of a periodic pulse train is introduced to all neurons forming the network, the stochastic resonance decreases as compared to the case when only a single neuron is paced. Moreover, we show that finite delays in coupling can significantly affect the stochastic resonance on scale-free neuronal networks. In particular, appropriately tuned delays can induce multiple stochastic resonances independently of the placing of the pacemaker, but they can also altogether destroy stochastic resonance. Delay-induced multiple stochastic resonances manifest as well-expressed maxima of the correlation measure, appearing at every multiple of the pacemaker period. We argue that fine-tuned delays and locally active pacemakers are vital for assuring optimal conditions for stochastic resonance on complex neuronal networks. PMID- 19566249 TI - Detrended fluctuation analysis of the magnetic and electric field variations that precede rupture. AB - Magnetic field variations are detected before rupture in the form of "spikes" of alternating sign. The distinction of these spikes from random noise is of major practical importance since it is easier to conduct magnetic field measurements than electric field ones. Applying detrended fluctuation analysis (DFA), these spikes look to be random at short time lags. On the other hand, long-range correlations prevail at time lags larger than the average time interval between consecutive spikes with a scaling exponent alpha around 0.9. In addition, DFA is applied to recent preseismic electric field variations in long duration (several hours to a couple of days) and reveals a scale invariant feature with an exponent alpha approximately 1 over all scales available (around five orders of magnitude). PMID- 19566248 TI - Effects of mucosal loading on vocal fold vibration. AB - A chain model was proposed in this study to examine the effects of mucosal loading on vocal fold vibration. Mucosal loading was defined as the loading caused by the interaction between the vocal folds and the surrounding tissue. In the proposed model, the vocal folds and the surrounding tissue were represented by a series of oscillators connected by a coupling spring. The lumped masses, springs, and dampers of the oscillators modeled the tissue properties of mass, stiffness, and viscosity, respectively. The coupling spring exemplified the tissue interactions. By numerically solving this chain model, the effects of mucosal loading on the phonation threshold pressure, phonation instability pressure, and energy distribution in a voice production system were studied. It was found that when mucosal loading is small, phonation threshold pressure increases with the damping constant R(r), the mass constant R(m), and the coupling constant R(mu) of mucosal loading but decreases with the stiffness constant R(k). Phonation instability pressure is also related to mucosal loading. It was found that phonation instability pressure increases with the coupling constant R(mu) but decreases with the stiffness constant R(k) of mucosal loading. Therefore, it was concluded that mucosal loading directly affects voice production. PMID- 19566250 TI - Singularities of two-dimensional invertible piecewise isometric dynamics. AB - We investigate the singularity structure of two-dimensional invertible piecewise isometric dynamics. The main purpose of this research is to establish the connection between the geometrical properties of the singularity and the dynamics of the system. We classify the singularity of two-dimensional bounded invertible piecewise isometric dynamics into three types with respect to their geometrical properties. Among the three, we show that one type of the singularity can be removed by lifting up the dynamics to a suitably defined (branched) manifold. Among the remaining two, we prove that only one of them contributes to the intricate orbit structure of the system and generates the sensitive dependence on the initial condition, while the other does nothing. PMID- 19566251 TI - Propagation failures, breathing fronts, and nonannihilation collisions in the ferroin-bromate-pyrocatechol system. AB - The emergence of propagating pulses was investigated with the photosensitive ferroin-bromate-pyrocatechol reaction in capillary tubes, in which various interesting spatiotemporal behaviors such as propagation failure, breathing fronts, and transitions between propagating pulses and fronts have been observed. Rather than a mutual annihilation, the collision of a propagating pulse and a growing front forces the front to recede gradually. A phase diagram in the pyrocatechol-bromate concentration space shows that the pulse instabilities take place throughout the conditions at which the system generates wave activities, suggesting that the presence of coupled autocatalytic feedbacks may facilitate the onset of pulse instabilities. PMID- 19566252 TI - Long time evolution of phase oscillator systems. AB - It is shown, under weak conditions, that the dynamical evolution of large systems of globally coupled phase oscillators with Lorentzian distributed oscillation frequencies is, in an appropriate physical sense, time-asymptotically attracted toward a reduced manifold of the system states. This manifold was previously known and used to facilitate the discovery of attractors and bifurcations of such systems. The result of this paper establishes that attractors for the order parameter dynamics obtained by restriction to this reduced manifold are, in fact, the only such attractors of the full system. Thus all long time dynamical behaviors of the order parameters of these systems can be obtained by restriction to the reduced manifold. PMID- 19566253 TI - Antiphase and in-phase synchronization of nonlinear oscillators: the Huygens's clocks system. AB - We introduce an interaction mechanism between oscillators leading to exact antiphase and in-phase synchronization. This mechanism is applied to the coupling between two nonlinear oscillators with a limit cycle in phase space, leading to a simple justification of the antiphase synchronization as observed in Huygens's pendulum clocks experiment. If the two coupled nonlinear oscillators reach the antiphase or the in-phase synchronized oscillatory state, the period of oscillation is different from the eigenperiods of the uncoupled oscillators. PMID- 19566254 TI - Complex modular structure of large-scale brain networks. AB - Modular structure is ubiquitous among real-world networks from related proteins to social groups. Here we analyze the modular organization of brain networks at a large scale (voxel level) extracted from functional magnetic resonance imaging signals. By using a random-walk-based method, we unveil the modularity of brain webs and show modules with a spatial distribution that matches anatomical structures with functional significance. The functional role of each node in the network is studied by analyzing its patterns of inter- and intramodular connections. Results suggest that the modular architecture constitutes the structural basis for the coexistence of functional integration of distant and specialized brain areas during normal brain activities at rest. PMID- 19566255 TI - Phase synchronization analysis by assessment of the phase difference gradient. AB - Phase synchronization is an important phenomenon of nonlinear dynamics and has recently received much scientific attention. In this work a method for identifying phase synchronization epochs is described which focuses on estimating the gradient of segments of the generalized phase differences between phase slips in an experimental time series. In phase synchronized systems, there should be a zero gradient of the generalized phase differences even if the systems are contaminated by noise. A method which tests if the gradient of the generalized phase difference is statistically different from zero is reported. The method has been validated by numerical studies on model systems and by comparing the results to those published previously. The method is applied to cardiorespiratory time series from a human volunteer measured in clinical settings and compared to synchrogram analysis for the same data. Potential problems with synchrogram analysis of experimental data are discussed. PMID- 19566256 TI - Adaptive control and synchronization in a class of partially unknown chaotic systems. AB - This paper first addresses a novel control scheme to control a class of chaotic systems. In this method, no knowledge on the bounds of perturbations and disturbances is required in advance and parameters of the proposed controller are updated through an adaptive algorithm. Using the Lyapunov theory is employed to guarantee the stability of the closed loop system. Then synchronization of two nonidentical uncertain chaotic systems is investigated. To demonstrate the feasibility of the proposed scheme, numerical simulations on the application of control/synchronization of some famous chaotic systems are provided. PMID- 19566257 TI - General anticipating response in coupled dynamical systems. AB - We suggest a general anticipating response scheme from which a large class of different drive-response systems can be constructed by choosing a function to be determined. A specific case of the general scheme is studied in detail. This specific scheme, which is called the accelerative anticipating response scheme, makes the response system evolve along the same trajectory as the driver but with a faster speed. This counterintuitive phenomenon is proved to be robust against the disturbances of the driving term and the driving signal as well as parameter mismatches. So it may have potential applications in the environment where disturbances and parameter mismatches are unavoidable. PMID- 19566258 TI - On the energy transfer to small scales in a discrete model of one-dimensional turbulence. AB - We consider a discrete one-dimensional model that was investigated numerically by Daumont and Peyrard [Chaos 13, 624 (2003)] as a model for turbulence in fluids, i.e., for the energy transfer from large to small scales. They found numerically that there exist two different regimes for the energy spectrum at high energies and low energies, respectively, and gave an analytical explanation for the high energy spectrum. An analytical explanation is given here for the low-energy spectrum, which corresponds to the laminar regime. PMID- 19566259 TI - On the dynamics of chaotic spiking-bursting transition in the Hindmarsh-Rose neuron. AB - The paper considers the neuron model of Hindmarsh-Rose and studies in detail the system dynamics which controls the transition between the spiking and bursting regimes. In particular, such a passage occurs in a chaotic region and different explanations have been given in the literature to represent the process, generally based on a slow-fast decomposition of the neuron model. This paper proposes a novel view of the chaotic spiking-bursting transition exploiting the whole system dynamics and putting in evidence the essential role played in the phenomenon by the manifolds of the equilibrium point. An analytical approximation is developed for the related crucial elements and a subsequent numerical analysis signifies the properness of the suggested conjecture. PMID- 19566260 TI - Estimation of the control parameter from symbolic sequences: unimodal maps with variable critical point. AB - The work described in this paper can be interpreted as an application of the order patterns of symbolic dynamics when dealing with unimodal maps. Specifically, it is shown how Gray codes can be used to estimate the probability distribution functions (PDFs) of the order patterns of unimodal maps whose dynamics is controlled by an external parameter. Furthermore, these PDFs depend on the value of the external parameter, which eventually provides a handle to estimate the parameter value from symbolic sequences (in form of Gray codes), even when the critical point depends on the parameter. PMID- 19566261 TI - Introduction to focus issue: bipedal locomotion--from robots to humans. AB - Running and walking, collectively referred to as bipedal locomotion, represent self-organized behaviors generated by a spatially distributed dynamical system operating under the constraint that a person must be able to move without falling down. The organizing principles involve both forces actively regulated by the nervous system and those generated passively by the biomechanical properties of the musculoskeletal system and the environment in which the movements occur. With the development of modern motion capture and electrophysiological techniques it has become possible to explore the dynamical interplay between the passive and active controllers of locomotion in a manner that directly compares observation to predictions made by relevant mathematical and computer models. Consequently, many of the techniques initially developed to study nonlinear dynamical systems, including stability analyses, phase resetting and entrainment properties of limit cycles, and fractal and multifractal analysis, have come to play major roles in guiding progress. This Focus Issue discusses bipedal locomotion from the point of view of dynamical systems theory with the goal of stimulating discussion between the dynamical systems, physics, biomechanics, and neuroscience communities. PMID- 19566262 TI - Neuromechanical considerations for incorporating rhythmic arm movement in the rehabilitation of walking. AB - We have extensively used arm cycling to study the neural control of rhythmic movements such as arm swing during walking. Recently rhythmic movement of the arms has also been shown to enhance and shape muscle activity in the legs. However, restricted information is available concerning the conditions necessary to maximally alter lumbar spinal cord excitability. Knowledge on the neuromechanics of a task can assist in the determination of the type, level, and timing of neural signals, yet arm swing during walking and arm cycling have not received a detailed neuromechanical comparison. The purpose of this research was to provide a combined neural and mechanical measurement approach that could be used to assist in the determination of the necessary and sufficient conditions for arm movement to assist in lower limb rehabilitation after stroke and spinal cord injury. Subjects performed three rhythmic arm movement tasks: (1) cycling (cycle); (2) swinging while standing (swing); and (3) swinging while treadmill walking (walk). We hypothesized that any difference in neural control between tasks (i.e., pattern of muscle activity) would reflect changes in the mechanical constraints unique to each task. Three-dimensional kinematics were collected simultaneously with force measurement at the hand and electromyography from the arms and trunk. All data were appropriately segmented to allow a comparison between and across conditions and were normalized and averaged to 100% movement cycle based on shoulder excursion. Separate mathematical principal components analysis of kinematic and neural variables was performed to determine common task features and muscle synergies. The results highlight important neural and mechanical features that distinguish differences between tasks. For example, there are considerable differences in the anatomical positions of the arms during each task, which relate to the moments experienced about the elbow and shoulder. Also, there are differences between tasks in elbow flexion/extension kinematics alongside differential muscle activation profiles. As well, mechanical assistance and constraints during all tasks could affect muscle recruitment and the functional role of muscles. Overall, despite neural and mechanical differences, the results are consistent with conserved common central motor control mechanisms operational for cycle, walk, and swing but appropriately sculpted to demands unique to each task. However, changing the mechanical parameters could affect the role of afferent feedback altering neural control and the coupling to the lower limbs. PMID- 19566263 TI - Dynamic stability and phase resetting during biped gait. AB - Dynamic stability during periodic biped gait in humans and in a humanoid robot is considered. Here gait systems of human neuromusculoskeletal system and a humanoid are simply modeled while keeping their mechanical properties plausible. We prescribe periodic gait trajectories in terms of joint angles of the models as a function of time. The equations of motion of the models are then constrained by one of the prescribed gait trajectories to obtain types of periodically forced nonlinear dynamical systems. Simulated gait of the models may or may not fall down during gait, since the constraints are made only for joint angles of limbs but not for the motion of the body trunk. The equations of motion can exhibit a limit cycle solution (or an oscillatory solution that can be considered as a limit cycle practically) for each selected gait trajectory, if an initial condition is set appropriately. We analyze the stability of the limit cycle in terms of Poincare maps and the basin of attraction of the limit cycle in order to examine how the stability depends on the prescribed trajectory. Moreover, the phase resetting of gait rhythm in response to external force perturbation is modeled. Since we always prescribe a gait trajectory in this study, reacting gait trajectories during the phase resetting are also prescribed. We show that an optimally prescribed reacting gait trajectory with an appropriate amount of the phase resetting can increase the gait stability. Neural mechanisms for generation and modulation of the gait trajectories are discussed. PMID- 19566264 TI - Nonlinear time series analysis of knee and ankle kinematics during side by side treadmill walking. AB - Nonlinear time series analysis was used to estimate maximal Lyapunov exponents of select ankle and knee kinematics during three different conditions of treadmill walking: independent, side by side, and side by side with forced synchronization of stepping. Stride to stride variability was significantly increased for the condition in which individuals walked side by side and synchronized unintentionally when compared to the conditions of forced synchronization and independent walking. In addition, standard deviations of three kinematic variables of lower extremity movement were significantly increased during the condition in which unintentional synchronization occurred. No relationship was found between standard deviation and estimates of maximal Lyapunov exponents. An increase in kinematic variability during side by side walking for nonimpaired individuals who are not at risk of falling suggests that variability in certain aspects of performance might be indicative of a healthy system. Modeling this variability for an impaired individual to imitate may have beneficial effects on locomotor function. These results may therefore have implications for the rehabilitation of gait in humans by suggesting that a different functional outcome might be achieved by practicing side by side walking as opposed to more commonly used strategies involving independent walking. PMID- 19566265 TI - Maneuvers during legged locomotion. AB - Maneuverability is essential for locomotion. For animals in the environment, maneuverability is directly related to survival. For humans, maneuvers such as turning are associated with increased risk for injury, either directly through tissue loading or indirectly through destabilization. Consequently, understanding the mechanics and motor control of maneuverability is a critical part of locomotion research. We briefly review the literature on maneuvering during locomotion with a focus on turning in bipeds. Walking turns can use one of several different strategies. Anticipation can be important to adjust kinematics and dynamics for smooth and stable maneuvers. During running, turns may be substantially constrained by the requirement for body orientation to match movement direction at the end of a turn. A simple mathematical model based on the requirement for rotation to match direction can describe leg forces used by bipeds (humans and ostriches). During running turns, both humans and ostriches control body rotation by generating fore-aft forces. However, whereas humans must generate large braking forces to prevent body over-rotation, ostriches do not. For ostriches, generating the lateral forces necessary to change movement direction results in appropriate body rotation. Although ostriches required smaller braking forces due in part to increased rotational inertia relative to body mass, other movement parameters also played a role. Turning performance resulted from the coordinated behavior of an integrated biomechanical system. Results from preliminary experiments on horizontal-plane stabilization support the hypothesis that controlling body rotation is an important aspect of stable maneuvers. In humans, body orientation relative to movement direction is rapidly stabilized during running turns within the minimum of two steps theoretically required to complete analogous maneuvers. During straight running and cutting turns, humans exhibit spring-mass behavior in the horizontal plane. Changes in the horizontal projection of leg length were linearly related to changes in horizontal-plane leg forces. Consequently, the passive dynamic stabilization associated with spring-mass behavior may contribute to stability during maneuvers in bipeds. Understanding the mechanics of maneuverability will be important for understanding the motor control of maneuvers and also potentially be useful for understanding stability. PMID- 19566267 TI - Neuromechanical models for insect locomotion: Stability, maneuverability, and proprioceptive feedback. AB - We describe a hierarchy of models for legged locomotion, emphasizing relationships among feedforward (preflexive) stability, maneuverability, and reflexive feedback. We focus on a hexapedal geometry representative of insect locomotion in the ground plane that includes a neural central pattern generator circuit, nonlinear muscles, and a representative proprioceptive sensory pathway. Although these components of the model are rather complex, neglect of leg mass yields a neuromechanical system with only three degrees of freedom, and numerical simulations coupled with a Poincare map analysis shows that the feedforward dynamics is strongly stable, apart from one relatively slow mode and a neutral mode in body yaw angle. These modes moderate high frequency perturbations, producing slow heading changes that can be corrected by a stride-to-stride steering strategy. We show that the model's response to a lateral impulsive perturbation closely matches that of a cockroach subject to a similar impulse. We also describe preliminary studies of proprioceptive leg force feedback, showing how a reflexive pathway can reinforce the preflexive stability inherent in the system. PMID- 19566268 TI - Understanding the complexity of human gait dynamics. AB - Time series of human gait stride intervals exhibit fractal and multifractal properties under several conditions. Records from subjects walking at normal, slow, and fast pace speed are analyzed to determine changes in the fractal scalings as a function of the stress condition of the system. Records from subjects with different age from children to elderly and patients suffering from neurodegenerative disease are analyzed to determine changes in the fractal scalings as a function of the physical maturation or degeneration of the system. A supercentral pattern generator model is presented to simulate the above two properties that are typically found in dynamical network performance: that is, how a dynamical network responds to stress and to evolution. PMID- 19566266 TI - Lateral stability of the spring-mass hopper suggests a two-step control strategy for running. AB - This paper investigates the control of running gaits in the context of a spring loaded inverted pendulum model in three dimensions. Specifically, it determines the minimal number of steps required for an animal to recover from a perturbation to a specified gait. The model has four control inputs per step: two touchdown angles (azimuth and elevation) and two spring constants (compression and decompression). By representing the locomotor movement as a discrete-time return map and using the implicit function theorem we show that the number of recovery steps needed following a perturbation depends upon the goals of the control mechanism. When the goal is to follow a straight line, two steps are necessary and sufficient for small lateral perturbations. Multistep control laws have a larger number of control inputs than outputs, so solutions of the control problem are not unique. Additional constraints, referred to here as synergies, are imposed to determine unique control inputs for perturbations. For some choices of synergies, two-step control can be expressed as two iterations of its first step policy and designed so that recovery occurs in just one step for all perturbations for which one-step recovery is possible. PMID- 19566270 TI - The time-delayed inverted pendulum: implications for human balance control. AB - The inverted pendulum is frequently used as a starting point for discussions of how human balance is maintained during standing and locomotion. Here we examine three experimental paradigms of time-delayed balance control: (1) mechanical inverted time-delayed pendulum, (2) stick balancing at the fingertip, and (3) human postural sway during quiet standing. Measurements of the transfer function (mechanical stick balancing) and the two-point correlation function (Hurst exponent) for the movements of the fingertip (real stick balancing) and the fluctuations in the center of pressure (postural sway) demonstrate that the upright fixed point is unstable in all three paradigms. These observations imply that the balanced state represents a more complex and bounded time-dependent state than a fixed-point attractor. Although mathematical models indicate that a sufficient condition for instability is for the time delay to make a corrective movement, tau(n), be greater than a critical delay tau(c) that is proportional to the length of the pendulum, this condition is satisfied only in the case of human stick balancing at the fingertip. Thus it is suggested that a common cause of instability in all three paradigms stems from the difficulty of controlling both the angle of the inverted pendulum and the position of the controller simultaneously using time-delayed feedback. Considerations of the problematic nature of control in the presence of delay and random perturbations ("noise") suggest that neural control for the upright position likely resembles an adaptive type controller in which the displacement angle is allowed to drift for small displacements with active corrections made only when theta exceeds a threshold. This mechanism draws attention to an overlooked type of passive control that arises from the interplay between retarded variables and noise. PMID- 19566269 TI - Control entropy identifies differential changes in complexity of walking and running gait patterns with increasing speed in highly trained runners. AB - Regularity statistics have been previously applied to walking gait measures in the hope of gaining insight into the complexity of gait under different conditions and in different populations. Traditional regularity statistics are subject to the requirement of stationarity, a limitation for examining changes in complexity under dynamic conditions such as exhaustive exercise. Using a novel measure, control entropy (CE), applied to triaxial continuous accelerometry, we report changes in complexity of walking and running during increasing speeds up to exhaustion in highly trained runners. We further apply Karhunen-Loeve analysis in a new and novel way to the patterns of CE responses in each of the three axes to identify dominant modes of CE responses in the vertical, mediolateral, and anterior/posterior planes. The differential CE responses observed between the different axes in this select population provide insight into the constraints of walking and running in those who may have optimized locomotion. Future comparisons between athletes, healthy untrained, and clinical populations using this approach may help elucidate differences between optimized and diseased locomotor control. PMID- 19566271 TI - Neuromechanical tuning of nonlinear postural control dynamics. AB - Postural control may be an ideal physiological motor task for elucidating general questions about the organization, diversity, flexibility, and variability of biological motor behaviors using nonlinear dynamical analysis techniques. Rather than presenting "problems" to the nervous system, the redundancy of biological systems and variability in their behaviors may actually be exploited to allow for the flexible achievement of multiple and concurrent task-level goals associated with movement. Such variability may reflect the constant "tuning" of neuromechanical elements and their interactions for movement control. The problem faced by researchers is that there is no one-to-one mapping between the task goal and the coordination of the underlying elements. We review recent and ongoing research in postural control with the goal of identifying common mechanisms underlying variability in postural control, coordination of multiple postural strategies, and transitions between them. We present a delayed-feedback model used to characterize the variability observed in muscle coordination patterns during postural responses to perturbation. We emphasize the significance of delays in physiological postural systems, requiring the modulation and coordination of both the instantaneous, "passive" response to perturbations as well as the delayed, "active" responses to perturbations. The challenge for future research lies in understanding the mechanisms and principles underlying neuromechanical tuning of and transitions between the diversity of postural behaviors. Here we describe some of our recent and ongoing studies aimed at understanding variability in postural control using physical robotic systems, human experiments, dimensional analysis, and computational models that could be enhanced from a nonlinear dynamics approach. PMID- 19566272 TI - Optimal speeds for walking and running, and walking on a moving walkway. AB - Many aspects of steady human locomotion are thought to be constrained by a tendency to minimize the expenditure of metabolic cost. This paper has three parts related to the theme of energetic optimality: (1) a brief review of energetic optimality in legged locomotion, (2) an examination of the notion of optimal locomotion speed, and (3) an analysis of walking on moving walkways, such as those found in some airports. First, I describe two possible connotations of the term "optimal locomotion speed:" that which minimizes the total metabolic cost per unit distance and that which minimizes the net cost per unit distance (total minus resting cost). Minimizing the total cost per distance gives the maximum range speed and is a much better predictor of the speeds at which people and horses prefer to walk naturally. Minimizing the net cost per distance is equivalent to minimizing the total daily energy intake given an idealized modern lifestyle that requires one to walk a given distance every day--but it is not a good predictor of animals' walking speeds. Next, I critique the notion that there is no energy-optimal speed for running, making use of some recent experiments and a review of past literature. Finally, I consider the problem of predicting the speeds at which people walk on moving walkways--such as those found in some airports. I present two substantially different theories to make predictions. The first theory, minimizing total energy per distance, predicts that for a range of low walkway speeds, the optimal absolute speed of travel will be greater--but the speed relative to the walkway smaller--than the optimal walking speed on stationary ground. At higher walkway speeds, this theory predicts that the person will stand still. The second theory is based on the assumption that the human optimally reconciles the sensory conflict between the forward speed that the eye sees and the walking speed that the legs feel and tries to equate the best estimate of the forward speed to the naturally preferred speed. This sensory conflict theory also predicts that people would walk slower than usual relative to the walkway yet move faster than usual relative to the ground. These predictions agree qualitatively with available experimental observations, but there are quantitative differences. PMID- 19566274 TI - Delayed and lasting effects of deep brain stimulation on locomotion in Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder characterized by a variety of motor signs affecting gait, postural stability, and tremor. These symptoms can be improved when electrodes are implanted in deep brain structures and electrical stimulation is delivered chronically at high frequency (>100 Hz). Deep brain stimulation (DBS) onset or cessation affects PD signs with different latencies, and the long-term improvements of symptoms affecting the body axis and those affecting the limbs vary in duration. Interestingly, these effects have not been systematically analyzed and modeled. We compare these timing phenomena in relation to one axial (i.e., locomotion) and one distal (i.e., tremor) signs. We suggest that during DBS, these symptoms are improved by different network mechanisms operating at multiple time scales. Locomotion improvement may involve a delayed plastic reorganization, which takes hours to develop, whereas rest tremor is probably alleviated by an almost instantaneous desynchronization of neural activity in subcortical structures. Even if all PD patients develop both distal and axial symptoms sooner or later, current computational models of locomotion and rest tremor are separate. Furthermore, a few computational models of locomotion focus on PD and none exploring the effect of DBS was found in the literature. We, therefore, discuss a model of a neuronal network during DBS, general enough to explore the subcircuits controlling locomotion and rest tremor simultaneously. This model accounts for synchronization and plasticity, two mechanisms that are believed to underlie the two types of symptoms analyzed. We suggest that a hysteretic effect caused by DBS-induced plasticity and synchronization modulation contributes to the different therapeutic latencies observed. Such a comprehensive, generic computational model of DBS effects, incorporating these timing phenomena, should assist in developing a more efficient, faster, durable treatment of distal and axial signs in PD. PMID- 19566275 TI - Comment on "Rewiring networks for synchronization" [Chaos 18, 037105 (2008)]. AB - This is a comment on a recent paper by A. Hagberg and D. A. Schult [Chaos 18, 037105 (2008)]. By taking the eigenratio of the Laplacian of an undirected and unweighted network as its synchronizability measure, they proposed a greedy rewiring algorithm for enhancing the synchronizability of the network. The algorithm is not capable of avoiding local minima, and as a consequence, for each initial network, different optimized networks with different synchronizabilities are obtained. Here, we show that by employing a simulated annealing based optimization method, it is possible to further enhance the synchronizability of the network. Moreover, using this approach, the optimized network is not biased by the initial network and regardless of the initial networks, the final optimized networks have similar synchronization properties. PMID- 19566276 TI - Introduction to controversial topics in nonlinear science: is the normal heart rate chaotic? AB - In June 2008, the editors of Chaos decided to institute a new section to appear from time to time that addresses timely and controversial topics related to nonlinear science. The first of these deals with the dynamical characterization of human heart rate variability. We asked authors to respond to the following questions: Is the normal heart rate chaotic? If the normal heart rate is not chaotic, is there some more appropriate term to characterize the fluctuations (e.g., scaling, fractal, multifractal)? How does the analysis of heart rate variability elucidate the underlying mechanisms controlling the heart rate? Do any analyses of heart rate variability provide clinical information that can be useful in medical assessment (e.g., in helping to assess the risk of sudden cardiac death)? If so, please indicate what additional clinical studies would be useful for measures of heart rate variability to be more broadly accepted by the medical community. In addition, as a challenge for analysis methods, PhysioNet [A. L. Goldberger et al., "PhysioBank, PhysioToolkit, and PhysioNet: Components of a new research resource for complex physiologic signals," Circulation 101, e215-e220 (2000)] provided data sets from 15 patients of whom five were normal, five had heart failure, and five had atrial fibrillation (http://www.physionet.org/challenge/chaos/). This introductory essay summarizes the main issues and introduces the essays that respond to these questions. PMID- 19566277 TI - Delays in the human heartbeat dynamics. AB - This paper explores the possibility of applying statistical nonlinear physics methods to elucidate the underlying mechanisms controlling the heart rate. In particular, the presence of delays in RR interval dynamics is studied by using a lagged detrended fluctuation analysis. The results indicate that patients with congestive heart failure (CHF) have a prolonged time delay in the baroreflex response. Some implications of large delays for the functioning of autonomic control in subjects with CHF are discussed. PMID- 19566273 TI - Gait dynamics in Parkinson's disease: common and distinct behavior among stride length, gait variability, and fractal-like scaling. AB - Parkinson's disease (PD) is a common, debilitating neurodegenerative disease. Gait disturbances are a frequent cause of disability and impairment for patients with PD. This article provides a brief introduction to PD and describes the gait changes typically seen in patients with this disease. A major focus of this report is an update on the study of the fractal properties of gait in PD, the relationship between this feature of gait and stride length and gait variability, and the effects of different experimental conditions on these three gait properties. Implications of these findings are also briefly described. This update highlights the idea that while stride length, gait variability, and fractal scaling of gait are all impaired in PD, distinct mechanisms likely contribute to and are responsible for the regulation of these disparate gait properties. PMID- 19566278 TI - Normal heartbeat series are nonchaotic, nonlinear, and multifractal: new evidence from semiparametric and parametric tests. AB - We present new evidence that normal heartbeat series are nonchaotic, nonlinear, and multifractal. In addition to considering the largest Lyapunov exponent and the correlation dimension, the results of the parametric and semiparametric estimation of the long memory parameter (long-range dependence) unambiguously reveal that the underlying process is nonstationary, multifractal, and has strong nonlinearity. PMID- 19566279 TI - On the nature of heart rate variability in a breathing normal subject: a stochastic process analysis. AB - Human heart rate is moderated by the autonomous nervous system acting predominantly through the sinus node (the main cardiac physiological pacemaker). One of the dominant factors that determine the heart rate in physiological conditions is its coupling with the respiratory rhythm. Using the language of stochastic processes, we analyzed both rhythms simultaneously taking the data from polysomnographic recordings of two healthy individuals. Each rhythm was treated as a sum of a deterministic drift term and a diffusion term (Kramers Moyal expansion). We found that normal heart rate variability may be considered as the result of a bidirectional coupling of two nonlinear oscillators: the heart itself and the respiratory system. On average, the diffusion (noise) component measured is comparable in magnitude to the oscillatory (deterministic) term for both signals investigated. The application of the Kramers-Moyal expansion may be useful for medical diagnostics providing information on the relation between respiration and heart rate variability. This interaction is mediated by the autonomous nervous system, including the baroreflex, and results in a commonly observed phenomenon--respiratory sinus arrhythmia which is typical for normal subjects and often impaired by pathology. PMID- 19566280 TI - Identifying chaos from heart rate: the right task? AB - Providing a conclusive answer to the question "is this dynamics chaotic?" remains very challenging when experimental data are investigated. We showed that such a task is actually a difficult problem in the case of heart rates. Nevertheless, an appropriate dynamical analysis can discriminate healthy subjects from patients. PMID- 19566281 TI - Characterizing heart rate variability by scale-dependent Lyapunov exponent. AB - Previous studies on heart rate variability (HRV) using chaos theory, fractal scaling analysis, and many other methods, while fruitful in many aspects, have produced much confusion in the literature. Especially the issue of whether normal HRV is chaotic or stochastic remains highly controversial. Here, we employ a new multiscale complexity measure, the scale-dependent Lyapunov exponent (SDLE), to characterize HRV. SDLE has been shown to readily characterize major models of complex time series including deterministic chaos, noisy chaos, stochastic oscillations, random 1/f processes, random Levy processes, and complex time series with multiple scaling behaviors. Here we use SDLE to characterize the relative importance of nonlinear, chaotic, and stochastic dynamics in HRV of healthy, congestive heart failure, and atrial fibrillation subjects. We show that while HRV data of all these three types are mostly stochastic, the stochasticity is different among the three groups. PMID- 19566282 TI - Multifractality and heart rate variability. AB - In this paper, we participate to the discussion set forth by the editor of Chaos for the controversy, "Is the normal heart rate chaotic?" Our objective was to debate the question, "Is there some more appropriate term to characterize the heart rate variability (HRV) fluctuations?" We focused on the approximately 24 h RR series prepared for this topic and tried to verify with two different techniques, generalized structure functions and wavelet transform modulus maxima, if they might be described as being multifractal. For normal and congestive heart failure subjects, the h(q) exponents showed to be decreasing for increasing q with both methods, as it should be for multifractal signals. We then built 40 surrogate series to further verify such hypothesis. For most of the series (approximately 75%-80% of cases) multifractality stood the test of the surrogate data employed. On the other hand, series coming from patients in atrial fibrillation showed a small, if any, degree of multifractality. The population analyzed is too small for definite conclusions, but the study supports the use of multifractal series to model HRV. Also it suggests that the regulatory action of autonomous nervous system might play a role in the observed multifractality. PMID- 19566283 TI - Is the normal heart rate "chaotic" due to respiration? AB - The incidence of cardiovascular diseases increases with the growth of the human population and an aging society, leading to very high expenses in the public health system. Therefore, it is challenging to develop sophisticated methods in order to improve medical diagnostics. The question whether the normal heart rate is chaotic or not is an attempt to elucidate the underlying mechanisms of cardiovascular dynamics and therefore a highly controversial topical challenge. In this contribution we demonstrate that linear and nonlinear parameters allow us to separate completely the data sets of the three groups provided for this controversial topic in nonlinear dynamics. The question whether these time series are chaotic or not cannot be answered satisfactorily without investigating the underlying mechanisms leading to them. We give an example of the dominant influence of respiration on heart beat dynamics, which shows that observed fluctuations can be mostly explained by respiratory modulations of heart rate and blood pressure (coefficient of determination: 96%). Therefore, we recommend reformulating the following initial question: "Is the normal heart rate chaotic?" We rather ask the following: "Is the normal heart rate 'chaotic' due to respiration?" PMID- 19566284 TI - Stochastic vagal modulation of cardiac pacemaking may lead to erroneous identification of cardiac "chaos". AB - Fluctuations in the time interval between two consecutive R-waves of electrocardiogram during normal sinus rhythm may result from irregularities in the autonomic drive of the pacemaking sinoatrial node (SAN). We use a biophysically detailed mathematical model of the action potentials of rabbit SAN to quantify the effects of fluctuations in acetylcholine (ACh) on the pacemaker activity of the SAN and its variability. Fluctuations in ACh concentration model the effect of stochastic activity in the vagal parasympathetic fibers that innervate the SAN and produce varying rates of depolarization during the pacemaker potential, leading to fluctuations in cycle length (CL). Both the estimated maximal Lyapunov exponent and the noise limit of the resultant sequence of fluctuating CLs suggest chaotic dynamics. Apparently chaotic heart rate variability (HRV) seen in sinus rhythm can be produced by stochastic modulation of the SAN. The identification of HRV data as chaotic by use of time series measures such as a positive maximal Lyapunov exponent or positive noise limit requires both caution and a quantitative, predictive mechanistic model that is fully deterministic. PMID- 19566285 TI - Imaging of glioma tumor with endogenous fluorescence tomography. AB - Tomographic imaging of a glioma tumor with endogenous fluorescence is demonstrated using a noncontact single-photon counting fan-beam acquisition system interfaced with microCT imaging. The fluorescence from protoporphyrin IX (PpIX) was found to be detectable, and allowed imaging of the tumor from within the cranium, even though the tumor presence was not visible in the microCT image. The combination of single-photon counting detection and normalized fluorescence to transmission detection at each channel allowed robust imaging of the signal. This demonstrated use of endogenous fluorescence stimulation from aminolevulinic acid (ALA) and provides the first in vivo demonstration of deep tissue tomographic imaging with protoporphyrin IX. PMID- 19566286 TI - Optically sectioned fluorescence endomicroscopy with hybrid-illumination imaging through a flexible fiber bundle. AB - We present an endomicroscope apparatus that exhibits out-of-focus background rejection based on wide-field illumination through a flexible imaging fiber bundle. Our technique, called HiLo microscopy, involves acquiring two images, one with grid-pattern illumination and another with standard uniform illumination. An evaluation of the image contrast with grid-pattern illumination provides an optically sectioned image with low resolution. This is complemented with high resolution information from the uniform illumination image, leading to a full resolution image that is optically sectioned. HiLo endomicroscope movies are presented of fluorescently labeled rat colonic mucosa. PMID- 19566287 TI - Intraluminal fluorescence spectroscopy catheter with ultrasound guidance. AB - We demonstrate the feasibility of a time-resolved fluorescence spectroscopy (TRFS) technique for intraluminal investigation of arterial vessel composition under intravascular ultrasound (IVUS) guidance. A prototype 1.8-mm (5.4 Fr) catheter combining a side-viewing optical fiber (SVOF) and an IVUS catheter was constructed and tested with in vitro vessel phantoms. The prototype catheter can locate a fluorophore in the phantom vessel wall, steer the SVOF in place, perform blood flushing under flow conditions, and acquire high-quality TRFS data using 337-nm wavelength excitation. The catheter steering capability used for the coregistration of the IVUS image plane and the SVOF beam produce a guiding precision to an arterial phantom wall site location of 0.53+/-0.16 mm. This new intravascular multimodal catheter enables the potential for in vivo arterial plaque composition identification using TRFS. PMID- 19566288 TI - In vivo trans-rectal ultrasound-coupled optical tomography of a transmissible venereal tumor model in the canine pelvic canal. AB - In vivo trans-rectal near-infrared (NIR) optical tomography was performed concurrently with, albeit reconstructed without spatial a prior of, trans-rectal ultrasound (US) on transmissible venereal tumor (TVT) developed as a model in the canine pelvic canal. Studies were taken longitudinally at prior to, 14 days after, and 35 days after the TVT injection. As the tumor grew, the nodules became increasingly hyperabsorptive and moderately hyperscattering on NIR. The regions of strong NIR contrast, especially on absorption images, correlated well with those of US hypoechoic masses indicative of tumors. Combining the information of trans-rectal NIR and US detected the tumor more accurately than did the US alone at 14 days postinjection. PMID- 19566289 TI - Developing compact multiphoton systems using femtosecond fiber lasers. AB - We implement a fiber-delivered compact femtosecond fiber laser at 1,030-nm wavelength in multiphoton imaging. The laser pulse duration is 150 fs, the average power is 200 mW, and the repetition rate is 40 MHz. The laser measures 200 x 160 x 45 mm in size and its output is delivered through a photonic bandgap fiber. Intrinsic second-harmonic generation signal is excited from rat tail tendon and human skin samples. Two-photon excited fluorescence signal is obtained from human skin tissues stained with exogenous fluorophore. Our results show that femtosecond fiber lasers at 1030-nm wavelength have significant potential in developing compact, all-fiber-based, portable multiphoton systems and endoscopes. PMID- 19566291 TI - Diffuse light suppression of back-directional gating imaging in high anisotropic media. AB - We experimentally demonstrate that back-directional gating in an imaging setup can potentially remove unwanted diffuse light to improve the contrast of an object embedded in a high anisotropic surrounding medium. In such back directional gating, the high anisotropic property of the surrounding medium can serve as a waveguide to deliver the incident light to the embedded object and to isolate the ballistic or snake-like light backscattered from the object in a moderate depth. We further discuss the effects of back-directional gating in the image formation in terms of the image resolution and the depth of field. Although backscattering detections of biological tissue have recently received considerable attention, we, for the first time to our knowledge, show its potential advantage for the contrast improvement in high anisotropic media. PMID- 19566290 TI - In-vivo imaging of murine tumors using complete-angle projection fluorescence molecular tomography. AB - We interrogate the ability of free-space fluorescence tomography to image small animals in vivo using charge-coupled device (CCD) camera measurements over 360 deg noncontact projections. We demonstrate the performance of normalized dual wavelength measurements that are essential for in-vivo use, as they account for the heterogeneous distribution of photons in tissue. In-vivo imaging is then showcased on mouse lung and brain tumors cross-validated by x-ray microcomputed tomography and histology. PMID- 19566292 TI - Simultaneous photoacoustic imaging technique using an acoustic imaging lens. AB - A simultaneous photoacoustic (PA) tomography imaging technique in multilayer samples was developed. Instead of using the PA image reconstruction methods on the basis of complex algorithms, obtaining a two-dimensional PA image in real time is available by using an acoustic lens that has the ability of parallel imaging. The imaging system can acquire the complete PA signals of high signal-to noise ratio from all the object planes by utilizing the advantages of the acoustic lens with long focal depth and a fast data acquisition system with a high sampling rate. With the time-resolved technique, the PA signals from different object planes can be distinguished and then the high optical contrast multilayer PA images can be reconstructed simultaneously without any algorithms. The experimental results show that the reconstructed sections agree well with the original samples. PMID- 19566293 TI - Speckle reduction in optical coherence tomography using angular compounding by B scan Doppler-shift encoding. AB - We propose a novel method for speckle reduction for optical coherence tomography based on angular compounding by B-scan Doppler-shift encoding (AngularCBD). By decentering the probe beam from the pivot of a scanning mirror, the illumination angle represented by different components of the beam can be encoded in Doppler shift. Compounding multiple images reconstructed from different Doppler-shift bands, we can suppress speckle without sacrificing image acquisition speed. Speckle reduction with AngularCBD is demonstrated by imaging a phantom and tissue sample in vitro and in vivo. PMID- 19566294 TI - Confocal microscopy of thick specimens. AB - Confocal microscopy is an excellent tool to gain structural information from deep within a biological sample. The depth from which information can be extracted as well as the resolution of the detection system are limited by spherical aberrations in the laser pathway. These spherical aberrations of the visible light can be efficiently canceled by optimizing the refractive index of the immersion media. Another way of cancelling spherical aberrations is by changing tube length, or alternatively, by changing the objective from infinite correction to finite correction, or vice versa, depending on which microscope is used. A combination of these two methods allows for confocal imaging at continuous depths. Presently, confocal microscopes typically operate at a maximum depth of 40 microm in the sample, but with the methods presented here, we show that information can easily be gained from depths up to 100 microm. Additionally, the precision of localization of a single fluorophore in the axial direction, limited by spherical aberrations, can be significantly improved, even if the fluorophore is located deep within the sample. In principle, this method can improve the efficiency of any kind of microscopy based on visible light. PMID- 19566295 TI - Influence of oxygen saturation on the optical scattering properties of human red blood cells in the spectral range 250 to 2,000 nm. AB - The intrinsic optical parameters absorption coefficient mu(a), scattering coefficient micros, anisotropy factor g, and effective scattering coefficient micros were determined for human red blood cell (RBC) suspensions of hematocrit 33.2% dependent on the oxygen saturation (SAT O(2)) in the wavelength range 250 to 2,000 nm, including the range above 1,100 nm, about which there are no data available in the literature. Integrating sphere measurements of light transmittance and reflectance in combination with inverse Monte Carlo simulation were carried out for SAT O(2) levels of 100 and 0%. In the wavelength range up to 1,200 nm, the absorption behavior is determined by the hemoglobin absorption. The spectral range above the cells' absorption shows no dependence on SAT O(2) and approximates the absorption of water with values 20 to 30% below the respective values for water. Parameters micros and g are significantly influenced by the SAT O(2)-induced absorption changes. Above 600 nm, micros decreases continuously from values of 85 mm(-1) to values of 30 mm(-1) at 2,000 nm. The anisotropy factor shows a slight decrease with wavelengths above 600 nm. In the spectral regions of 1,450 and 1,900 nm where water has local absorption maxima, g shows a significant decrease down to 0.85, whereas micros increases. PMID- 19566296 TI - Effect of low-level laser treatment of tissue-engineered skin substitutes: contraction of collagen lattices. AB - Fibroblast-populated collagen lattices (FPCL) are widely used in tissue engineered artificial skin substitutes, but their main drawback is that interaction of fibroblasts and matrix causes contraction of the lattice, reducing it to about 20% of its original area. The effect of low-level laser treatment (LLLT) on the behavior of 3T3 fibroblasts seeded in collagen lattices containing 20% chondroitin-6-sulphate was investigated to determine whether LLLT could control the contraction of FPCL. A He-Ne laser was used at 632.8 nm to deliver a 5-mW continuous wave with fluences from 1 to 4 J/cm(2). Laser treatment at 3 J/cm(2) increased contraction of collagen lattices in the absence of cells but decreased contraction of cell seeded lattices over a 7-day period. The effect was energy dependent and was not observed at 1, 2, or 4 J/cm(2). There was no alteration in fibroblast viability, morphology, or mitochondrial membrane potential after any laser treatments, but the distribution of actin fibers within the cells and collagen fibers in the matrices was disturbed at 3 J/cm(2). These effects contribute to the decrease in contraction observed. LLLT may offer a means to control contraction of FPCL used as artificial skin substitutes. PMID- 19566297 TI - Wavelet minimum description length detrending for near-infrared spectroscopy. AB - Near-infrared spectroscopy (NIRS) can be employed to investigate brain activities associated with regional changes of the oxy- and deoxyhemoglobin concentration by measuring the absorption of near-infrared light through the intact skull. NIRS is regarded as a promising neuroimaging modality thanks to its excellent temporal resolution and flexibility for routine monitoring. Recently, the general linear model (GLM), which is a standard method for functional MRI (fMRI) analysis, has been employed for quantitative analysis of NIRS data. However, the GLM often fails in NIRS when there exists an unknown global trend due to breathing, cardiac, vasomotion, or other experimental errors. We propose a wavelet minimum description length (Wavelet-MDL) detrending algorithm to overcome this problem. Specifically, the wavelet transform is applied to decompose NIRS measurements into global trends, hemodynamic signals, and uncorrelated noise components at distinct scales. The minimum description length (MDL) principle plays an important role in preventing over- or underfitting and facilitates optimal model order selection for the global trend estimate. Experimental results demonstrate that the new detrending algorithm outperforms the conventional approaches. PMID- 19566299 TI - Experimental and theoretical evaluation of rotating orthogonal polarization imaging. AB - Rotating orthogonal polarization imaging is a new technique that provides quantitative measurements of the polarization properties of scattering media, such as tissue, which are free from surface reflections. The technique is investigated using both experiments and Monte Carlo simulations of a polarizing target embedded within a scattering medium. The technique is sensitive to the polarization properties of the target up to a depth of 17 mean free paths. Preliminary images of bovine tendon, lamb tendon, chicken breast, and human skin are also demonstrated. PMID- 19566298 TI - Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning. AB - A multiphoton endoscopy system has been developed using a two-axis microelectromechanical systems (MEMS) mirror and double-cladding photonic crystal fiber (DCPCF). The MEMS mirror has a 2-mm-diam, 20-deg optical scanning angle, and 1.26-kHz and 780-Hz resonance frequencies on the x and y axes. The maximum number of resolvable focal spots of the MEMS scanner is 720 x 720 on the x and y axes, which indicates that the MEMS scanner can potentially support high resolution multiphoton imaging. The DCPCF is compared with standard single-mode fiber and hollow-core photonic bandgap fiber on the basis of dispersion, attenuation, and coupling efficiency properties. The DCPCF has high collection efficiency, and its dispersion can be compensated by grating pairs. Three configurations of probe design are investigated, and their imaging quality and field of view are compared. A two-lens configuration with a collimation and a focusing lens provides the optimum imaging performance and packaging flexibility. The endoscope is applied to image fluorescent microspheres and bovine knee joint cartilage. PMID- 19566300 TI - Dynamics of retinal photocoagulation and rupture. AB - In laser retinal photocoagulation, short (<20 ms) pulses have been found to reduce thermal damage to the inner retina, decrease treatment time, and minimize pain. However, the safe therapeutic window (defined as the ratio of power for producing a rupture to that of mild coagulation) decreases with shorter exposures. To quantify the extent of retinal heating and maximize the therapeutic window, a computational model of millisecond retinal photocoagulation and rupture was developed. Optical attenuation of 532-nm laser light in ocular tissues was measured, including retinal pigment epithelial (RPE) pigmentation and cell-size variability. Threshold powers for vaporization and RPE damage were measured with pulse durations ranging from 1 to 200 ms. A finite element model of retinal heating inferred that vaporization (rupture) takes place at 180-190 degrees C. RPE damage was accurately described by the Arrhenius model with activation energy of 340 kJ/mol. Computed photocoagulation lesion width increased logarithmically with pulse duration, in agreement with histological findings. The model will allow for the optimization of beam parameters to increase the width of the therapeutic window for short exposures. PMID- 19566302 TI - Lipid concentrations in human coronary artery determined with high wavenumber Raman shifted light. AB - Raman spectroscopy is a rapid nondestructive technique capable of assaying chemicals in human artery tissues and characterizing atherosclerotic plaques in vivo, but clinical applications through optical fiber-based catheters have been hindered by large background signals generated within the fibers. Previous workers realized that this background was reduced significantly in the high wavenumber (HWVN) Raman region ( approximately 2,400 cm(-1) to approximately 3,800 cm(-1)), and with proper selection of optical fibers, one could collect quality Raman spectra remotely via a single optical fiber with no additional filters or optics. This study compared lipid concentrations in coronary artery tissue that were determined with chemical assay techniques to those estimated from HWVN Raman spectra collected through a single optical fiber. The standard error of predictions between the Raman and chemical assay techniques were small for cholesterol and esterified cholesterols, at 1.2% and 2.7%, respectively. PMID- 19566301 TI - Enhanced detection of early-stage oral cancer in vivo by optical coherence tomography using multimodal delivery of gold nanoparticles. AB - Contrast in optical coherence tomography (OCT) images can be enhanced by utilizing surface plasmon resonant gold nanoparticles. To improve the poor in vivo transport of gold nanoparticles through biological barriers, an efficient delivery strategy is needed. In this study, the improved penetration and distribution of gold nanoparticles were achieved by microneedle and ultrasound, respectively, and it was demonstrated that this multimodal delivery of antibody conjugated PEGylated gold nanoparticles enhanced the contrast in in vivo OCT images of oral dysplasia in a hamster model. PMID- 19566303 TI - Measurement of the refractive index of human teeth by optical coherence tomography. AB - We describe a novel method based on optical coherence tomography (OCT) for the accurate measurement of the refractive index of in vitro human teeth. We obtain the refractive indices of enamel, dentin, and cementum to be 1.631+/-0.007, 1.540+/-0.013, and 1.582+/-0.010, respectively. The profile of the refractive index is readily obtained via an OCT B scan across a tooth. This method can be used to study the refractive index changes caused by dental decay and therefore has great potential for the clinical diagnosis of early dental caries. PMID- 19566304 TI - In situ permeation study of drug through the stratum corneum using attenuated total reflectance [corrected] Fourier transform infrared spectroscopic imaging. AB - Infrared (IR) spectroscopy is one of the most chemically specific analytical methods that gives information about composition, structure, and interactions in a material. IR spectroscopy has been successfully applied to study the permeation of xenobiotics through the skin. Combining IR spectroscopy with an IR array detector led to the development of Fourier transform infrared (FTIR) spectroscopic imaging, which generates chemical information from different areas of a sample at the microscopic level. This is particularly important for heterogeneous samples, such as skin. Attenuated total reflectance [corrected] (ATR)-FTIR imaging has been applied to measure, in situ, the diffusion of benzyl nicotinate (BN) through the outer layer of human skin [stratum corneum (SC)]. In vitro experiments have demonstrated the heterogeneous distribution of SC surface lipids before the penetration of a saturated solution of BN. Image analysis demonstrated a strong correlation between the distribution of lipids and drugs, while ethanol appeared to be homogenously distributed in the SC. These results show the ability of ATR-FTIR imaging to measure simultaneously the affinities of drug and solvent to the lipid-rich and lipid-poor skin domains, respectively, during permeation. This information may be useful in better understanding drug diffusion pathways through the SC. PMID- 19566305 TI - Sensitivity and specificity for detecting basal cell carcinomas in Mohs excisions with confocal fluorescence mosaicing microscopy. AB - Recent studies have demonstrated the ability of confocal fluorescence mosaicing microscopy to rapidly detect basal cell carcinomas (BCCs) directly in thick and fresh Mohs surgical excisions. Mosaics of confocal images display large areas of tissue with high resolution and magnification equivalent to 2x, which is the standard magnification when examining pathology. Comparison of mosaics to Mohs frozen histopathology was shown to be excellent for all types of BCCs. However, comparisons in the previous studies were visual and qualitative. In this work, we report the results of a semiquantitative preclinical study in which 45 confocal mosaics are blindly evaluated for the presence (or absence) of BCC tumor. The evaluations are made by two clinicians: a senior Mohs surgeon with prior expertise in interpreting confocal images, and a novice Mohs fellow with limited experience. The blinded evaluation is compared to the gold standard of frozen histopathology. BCCs are detected with an overall sensitivity of 96.6%, specificity of 89.2%, positive predictive value of 93.0%, and negative predictive value of 94.7%. The results demonstrate the potential clinical utility of confocal mosaicing microscopy toward rapid surgical pathology at the bedside to expedite and guide surgery. PMID- 19566306 TI - Raman spectroscopy of synovial fluid as a tool for diagnosing osteoarthritis. AB - For many years, viscosity has been the primary method used by researchers in rheumatology to assess the physiochemical properties of synovial fluid in both normal and osteoarthritic patients. However, progress has been limited by the lack of methods that provide multiple layers of information, use small sample volumes, and are rapid. Raman spectroscopy was used to assess the biochemical composition of synovial fluid collected from 40 patients with clinical evidence of knee osteoarthritis (OA) at the time of elective surgical treatment. Severity of knee osteoarthritis was assessed by a radiologist using Kellgren/Lawrence (K/L) scores from knee joint x rays, while light microscopy and Raman spectroscopy were used to examine synovial fluid (SF) aspirates (2 to 10 microL), deposited on fused silica slides. We show that Raman bands used to describe protein secondary structure and content can be used to detect changes in synovial fluid from osteoarthritic patients. Several Raman band intensity ratios increased significantly in spectra collected from synovial fluid in patients with radiological evidence of moderate-to-severe osteoarthritis damage. These ratios can be used to provide a "yes/no" damage assessment. These studies provide evidence that Raman spectroscopy would be a suitable candidate in the evaluation of joint damage in knee osteoarthritis patients. PMID- 19566307 TI - Doppler optical coherence tomography imaging of local fluid flow and shear stress within microporous scaffolds. AB - Establishing a relationship between perfusion rate and fluid shear stress in a 3D cell culture environment is an ongoing and challenging task faced by tissue engineers. We explore Doppler optical coherence tomography (DOCT) as a potential imaging tool for in situ monitoring of local fluid flow profiles inside porous chitosan scaffolds. From the measured fluid flow profiles, the fluid shear stresses are evaluated. We examine the localized fluid flow and shear stress within low- and high-porosity chitosan scaffolds, which are subjected to a constant input flow rate of 0.5 ml min(-1). The DOCT results show that the behavior of the fluid flow and shear stress in micropores is strongly dependent on the micropore interconnectivity, porosity, and size of pores within the scaffold. For low-porosity and high-porosity chitosan scaffolds examined, the measured local fluid flow and shear stress varied from micropore to micropore, with a mean shear stress of 0.49+/-0.3 dyn cm(-2) and 0.38+/-0.2 dyn cm(-2), respectively. In addition, we show that the scaffold's porosity and interconnectivity can be quantified by combining analyses of the 3D structural and flow images obtained from DOCT. PMID- 19566308 TI - Diffuse optical monitoring of hemodynamic changes in piglet brain with closed head injury. AB - We used a nonimpact inertial rotational model of a closed head injury in neonatal piglets to simulate the conditions following traumatic brain injury in infants. Diffuse optical techniques, including diffuse reflectance spectroscopy and diffuse correlation spectroscopy (DCS), were used to measure cerebral blood oxygenation and blood flow continuously and noninvasively before injury and up to 6 h after the injury. The DCS measurements of relative cerebral blood flow were validated against the fluorescent microsphere method. A strong linear correlation was observed between the two techniques (R=0.89, p<0.00001). Injury-induced cerebral hemodynamic changes were quantified, and significant changes were found in oxy- and deoxy-hemoglobin concentrations, total hemoglobin concentration, blood oxygen saturation, and cerebral blood flow after the injury. The diffuse optical measurements were robust and also correlated well with recordings of vital physiological parameters over the 6-h monitoring period, such as mean arterial blood pressure, arterial oxygen saturation, and heart rate. Finally, the diffuse optical techniques demonstrated sensitivity to dynamic physiological events, such as apnea, cardiac arrest, and hypertonic saline infusion. In total, the investigation corraborates potential of the optical methods for bedside monitoring of pediatric and adult human patients in the neurointensive care unit. PMID- 19566309 TI - Morphological studies of living cells using gold nanoparticles and dark-field optical section microscopy. AB - The morphologic changes of living cells under drug interactions were studied by using 80-nm gold nanoparticles and dark-field optical section microscopy. The gold nanoparticles were coated with poly (L-lysine), which attached to the membranes of various cells by way of electrostatic attractive force. A three dimensional (3-D) morphological image was obtained by measuring the peak scattering intensities of gold nanoparticles at different focal planes. An algorithm for the reconstruction of 3-D cell morphology was presented. With the measured nanoparticle images and calculations, we show morphologic changes of lung cancer cells under the interaction of cytochalasin D drug at different times. PMID- 19566310 TI - New cross-talk measure of near-infrared spectroscopy and its application to wavelength combination optimization. AB - In near-infrared spectroscopy, concentration changes in oxygenated and deoxygenated hemoglobin are calculated from the changes in the attenuation of the measurement light. This is done by solving a linear equation based on the modified Lambert-Beer law. To solve this equation, we need to know the partial optical pathlengths in the activated region in the brain. Because they are difficult to know, a wavelength-independent constant or a wavelength-dependent total optical pathlength has been substituted for these values in actual measurements. This kind of substitution inevitably produces errors, called cross talk, when calculating concentration changes. In this paper, we propose a new cross-talk measure for dual and triple wavelength measurements, and analyze it over various wavelength combinations. The results indicate that constant substitution is not inferior to total path-length substitution in dual wavelength measurements, and that total path-length substitution is very effective for triple wavelength measurements. PMID- 19566312 TI - Dual-spectrum laser source based on fiber continuum generation for integrated optical coherence and multiphoton microscopy. AB - A single-laser dual-spectrum source designed for integrated optical coherence and multiphoton microscopy is demonstrated. The source implements the laser characteristics needed to optimally perform both modalities while extending the spectral range for this imaging technique. It consists of a widely tunable, mode locked, Ti-sapphire laser with a portion of its output spectrally broadened via continuum generation in a photonic crystal fiber. The continuum-broadened beam allows for enhanced optical sectioning with optical coherence microscopy, while the unbroadened beam from the ultrashort-pulse Ti-sapphire laser optimally excites fluorescent markers. The noise power of the continuum-broadened beam is less than 1.1 dBmHz higher than the Ti-sapphire laser in the range from 1 Hz to 25 MHz, and the fiber shows no sign of damage after approximately 100 h of use. We demonstrate the use of this source across a wide spectral range by imaging green fluorescent protein-transfected mouse fibroblast cells costained with fluorescent dyes that are maximally excited at various wavelengths. Images of unstained in vivo human skin are also presented. This source extends the feasibility of this integrated imaging modality and will facilitate new investigations in in vivo microscopy, tissue engineering, and cell biology. PMID- 19566311 TI - Single-walled carbon nanotubes as a multimodal-thermoacoustic and photoacoustic contrast agent. AB - We have developed a novel carbon nanotube-based contrast agent for both thermoacoustic and photoacoustic tomography. In comparison to deionized water, single-walled carbon nanotubes exhibited more than twofold signal enhancement for thermoacoustic tomography at 3 GHz. In comparison to blood, they exhibited more than sixfold signal enhancement for photoacoustic tomography at 1064 nm wavelength. The large contrast enhancement of single-walled carbon nanotubes was further corroborated by tissue phantom imaging studies. PMID- 19566313 TI - Fabrication of indocyanine green encapsulated biodegradable microbubbles for structural and functional imaging of cancer. AB - We developed a novel dual-modal contrast agent for the structural and functional imaging of cancer. The contrast agent was fabricated by encapsulating indocyanine green (ICG) in poly(lactic-co-glycolic acid) (PLGA) microbubbles using a modified double-emulsion method. More stabilized absorption and fluorescence emission characteristics were observed for aqueous and plasma suspensions of ICG encapsulated microbubbles. The technical feasibility of concurrent structural and functional imaging was demonstrated through a series of benchtop tests in which the aqueous suspension of ICG-encapsulated microbubbles was injected into a transparent tube embedded in an Intralipid phantom at different flow rates and concentrations. Concurrent fluorescence imaging and B-mode ultrasound imaging successfully captured the changes of microbubble flow rate and concentration with high linearity and accuracy. One potential application of the proposed ICG encapsulated PLGA microbubbles is for the identification and characterization of peritumoral neovasculature for enhanced coregistration between tumor structural and functional boundaries in ultrasound-guided near-infrared diffuse optical tomography. PMID- 19566314 TI - Compensation of optical heterogeneity-induced artifacts in fluorescence molecular tomography: theory and in vivo validation. AB - We present a method for reduction of image artifacts induced by the optical heterogeneities of tissue in fluorescence molecular tomography (FMT) through identification and compensation of image regions that evidence propagation of emission light through thin or low-absorption tunnels in tissue. The light tunneled as such contributes to the emission image as spurious components that might substantially overwhelm the desirable fluorescence emanating from the targeted lesions. The proposed method makes use of the strong spatial correlation between the emission and excitation images to estimate the tunneled components and yield a residual image that mainly consists of the signal due to the desirable fluorescence. This residual image is further refined using a coincidence mask constructed for each excitation-emission image pair. The coincidence mask is essentially a map of the "hot spots" that occur in both excitation and emission images, as such areas are often associated with tunneled emission. In vivo studies are performed on a human colon adenocarcinoma xenograft tumor model with subcutaneous tumors and a murine breast adenocarcinoma model with aggressive tumor cell metastasis and growth in the lungs. Results demonstrate significant improvements in the reconstructions achieved by the proposed method. PMID- 19566315 TI - In vivo quantification of chromophore concentration using fluorescence differential path length spectroscopy. AB - We present an optical method based on fluorescence spectroscopy for measuring chromophore concentrations in vivo. Fluorescence differential path length spectroscopy (FPDS) determines chromophore concentration based on the fluorescence intensity corrected for absorption. The concentration of the photosensitizer m-THPC (Foscan) was studied in vivo in normal rat liver, which is highly vascularized and therefore highly absorbing. Concentration estimates of m THPC measured by FDPS on the liver are compared with chemical extraction. Twenty five rats were injected with 0.3 mg kg m-THPC. In vivo optical concentration measurements were performed on tissue 3, 24, 48, and 96 h after m-THPC administration to yield a 10-fold variation in tissue concentration. After the optical measurements, the liver was harvested for chemical extraction. FDPS showed good correlation with chemical extraction. FDPS also showed a correlation between m-THPC fluorescence and blood volume fraction at the two shortest drug light intervals. This suggests different compartmental localization of m-THPC for different drug-light intervals that can be resolved using fluorescence spectroscopy. Differences in measured m-THPC concentration between FDPS and chemical extraction are related to the interrogation volume of each technique; approximately 0.2 mm(3) and approximately 10(2) mm(3), respectively. This indicates intra-animal variation in m-THPC distribution in the liver on the scale of the FDPS sampling volume. PMID- 19566317 TI - Integrated microscopy for real-time imaging of mechanotransduction studies in live cells. AB - Mechanical force is an important stimulus and determinant of many vascular smooth muscle cell functions including contraction, proliferation, migration, and cell attachment. Transmission of force from outside the cell through focal adhesions controls the dynamics of these adhesion sites and initiates intracellular signaling cascades that alter cellular behavior. To understand the mechanism by which living cells sense mechanical forces, and how they respond and adapt to their environment, a critical first step is to develop a new technology to investigate cellular behavior at subcellular level that integrates an atomic force microscope (AFM) with total internal reflection fluorescence (TIRF) and fast-spinning disk (FSD) confocal microscopy, providing high spatial and temporal resolution. AFM uses a nanosensor to measure the cell surface topography and can apply and measure mechanical force with high precision. TIRF microscopy is an optical imaging technique that provides high-contrast images with high z resolution of fluorescently labeled molecules in the immediate vicinity of the cell-coverslip interface. FSD confocal microscopy allows rapid 3-D imaging throughout the cell in real time. The integrated system is broadly applicable across a wide range of molecular dynamic studies in any adherent live cells, allowing direct optical imaging of cell responses to mechanical stimulation in real time. PMID- 19566316 TI - Optoelectronic holographic otoscope for measurement of nano-displacements in tympanic membranes. AB - Current methodologies for characterizing tympanic membrane (TM) motion are usually limited to either average acoustic estimates (admittance or reflectance) or single-point mobility measurements, neither of which suffices to characterize the detailed mechanical response of the TM to sound. Furthermore, while acoustic and single-point measurements may aid in diagnosing some middle-ear disorders, they are not always useful. Measurements of the motion of the entire TM surface can provide more information than these other techniques and may be superior for diagnosing pathology. We present advances in our development of a new compact optoelectronic holographic otoscope (OEHO) system for full field-of-view characterization of nanometer-scale sound-induced displacements of the TM surface at video rates. The OEHO system consists of a fiber optic subsystem, a compact otoscope head, and a high-speed image processing computer with advanced software for recording and processing holographic images coupled to a computer-controlled sound-stimulation and recording system. A prototype OEHO system is in use in a medical research environment to address basic science questions regarding TM function. The prototype provides real-time observation of sound-induced TM displacement patterns over a broad frequency range. Representative time-averaged and stroboscopic holographic interferometry results in animals and human cadaver samples are shown, and their potential utility is discussed. PMID- 19566318 TI - Comparison of instruments for investigation of microcirculatory blood flow and red blood cell concentration. AB - The use of laser Doppler perfusion imaging (LDPI) and laser speckle perfusion imaging (LSPI) is well known in the noninvasive investigation of microcirculatory blood flow. This work compares the two techniques with the recently developed tissue viability (TiVi) imaging system, which is proposed as a useful tool to quantify red blood cell concentration in microcirculation. Three systems are evaluated with common skin tests such as the use of vasodilating and vasoconstricting drugs (methlynicotinate and clobetasol, respectively) and a reactive hyperaemia maneuver (using a sphygmomanometer). The devices investigated are the laser Doppler line scanner (LDLS), the laser speckle perfusion imager (FLPI)-both from Moor Instruments (Axminster, United Kingdom)-and the TiVi imaging system (WheelsBridge AB, Linkoping, Sweden). Both imaging and point scanning by the devices are used to quantify the provoked reactions. Perfusion images of vasodilatation and vasoconstriction are acquired with both LDLS and FLPI, while TiVi images are acquired with the TiVi imager. Time acquisitions of an averaged region of interest are acquired for temporal studies such as the reactive hyperaemia. In contrast to the change in perfusion over time with pressure, the TiVi imager shows a different response due its measurement of blood concentration rather than perfusion. The responses can be explained by physiological understanding. Although the three devices sample different compartments of tissue, and output essentially different variables, comparisons can be seen between the three systems. The LDLS system proves to be suited to measurement of perfusion in deeper vessels, while FLPI and TiVi showed sensitivity to more superficial nutritional supply. LDLS and FLPI are insensitive to the action of the vasoconstrictor, while TiVi shows the clear boundaries of the reaction. Assessment of the resolution, penetration depth, and acquisition rate of each instrument show complimentary features that should be taken into account when choosing a system for a particular clinical measurement. PMID- 19566319 TI - Diffuse reflectance spectroscopy characterization of hemoglobin and intralipid solutions: in vitro measurements with continuous variation of absorption and scattering. AB - Absorption and scattering processes in biological tissues are studied through reflectance spectroscopy in tissue-like phantoms. For this aim, an experimental setup is designed to independently control both processes in hemoglobin and intralipid solutions. From the analysis of the obtained spectra, a simple empirical power law equation is found that relates absorbance with scattering and absorption coefficients. This relationship includes three wavelength independent parameters, which can be determined geometry from in vitro measurements for each particular optical optode. The dependence of the optical path length on the absorption and scattering coefficients is also analyzed, and estimations of this parameter for physiological conditions are presented. This study is useful to better understand the scattering phenomena in biological tissue, and to obtain absolute concentration of absorber particles when a homogeneous medium can be assumed. PMID- 19566320 TI - In-vivo Fourier domain optical coherence tomography as a new tool for investigation of vasodynamics in the mouse model. AB - In-vivo imaging of the vascular system can provide novel insight into the dynamics of vasoconstriction and vasodilation. Fourier domain optical coherence tomography (FD-OCT) is an optical, noncontact imaging technique based on interferometry of short-coherent near-infrared light with axial resolution of less than 10 microm. In this study, we apply FD-OCT as an in-vivo imaging technique to investigate blood vessels in their anatomical context using temporally resolved image stacks. Our chosen model system is the murine saphenous artery and vein, due to their small inner vessel diameters, sensitive response to vasoactive stimuli, and advantageous anatomical position. The vascular function of male wild-type mice (C57BL/6) is determined at the ages of 6 and 20 weeks. Vasoconstriction is analyzed in response to dermal application of potassium (K(+)), and vasodilation in response to sodium nitroprusside (SNP). Vasodynamics are quantified from time series (75 sec, 4 frames per sec, 330 x 512 pixels per frame) of cross sectional images that are analyzed by semiautomated image processing software. The morphology of the saphenous artery and vein is determined by 3-D image stacks of 512 x 512 x 512 pixels. Using the FD-OCT technique, we are able to demonstrate age-dependent differences in vascular function and vasodynamics. PMID- 19566322 TI - Glucose concentration monitoring using a small Fabry-Perot etalon. AB - Accurate measurements of aqueous glucose concentrations have been made in a double-chamber Fabry-Perot etalon that can be miniaturized for subcutaneous implantation to determine the concentration of glucose in interstitial fluid. In general, optical approaches to glucose detection measure light intensity, which in tissue varies due to inherent scattering and absorption. In our measurements, we compare the spectral positions of transmission maximums in two adjunct sections of an etalon in order to determine the refractive index difference between these sections and therefore we can tolerate large changes in intensity. With this approach, we were able to determine aqueous glucose concentrations between 0 mg/dl and 700 mg/dl within the precision of our reference measurement (+/-2.5 mg/dl or 2% of the measurement value). The use of reference cavities eliminates interference due to temperature variations, and we show the temperature independence over a temperature range of 32 degrees C to 42 degrees C. Furthermore, external filters eliminate interference from large molecule contaminants. PMID- 19566321 TI - Use of an oxygen-carrying blood substitute to improve intravascular optical coherence tomography imaging. AB - Optical coherence tomography (OCT) is a catheter-based imaging technology with powerful resolution capable of identifying vulnerable plaques and guiding coronary intervention. However, a significant limitation of intravascular OCT imaging is its attenuation by blood. We propose that the use of an oxygen carrying blood substitute could potentially optimize OCT image quality. Surgical isolation of the descending thoracic aorta of six rabbits is performed, followed by intravascular OCT imaging of the abdominal aorta. Perfluorodecalin (PFD) is oxygenated using a bubble-through technique with 100% oxygen. OCT imaging is performed and compared using three different flushing modalities: PFD; saline; and blood. OCT imaging of the rabbit abdominal aorta is successful in all of the subjects. In each of the six studied subjects, flushing with PFD consistently provides dramatically better imaging of the vessel wall tissue structures. OCT image quality is highly dependent on the ability of the flushing modality to remove blood from the imaging field. From this proof-of-concept study, we demonstrate that endovascular flushing with an oxygen-carrying blood substitute (PFD) is optically superior to saline flushing for intravascular imaging. PMID- 19566323 TI - High heterogeneity of plasma membrane microfluidity in multidrug-resistant cancer cells. AB - Diffusion-time distribution analysis (DDA) has been used to explore the plasma membrane fluidity of multidrug-resistant cancer cells (LR73 carcinoma cells) and also to characterize the influence of various membrane agents present in the extracellular medium. DDA is a recent single-molecule technique, based on fluorescence correlation spectroscopy (FCS), well suited to retrieve local organization of cell membrane. The method was conducted on a large number of living cells, which enabled us to get a detailed overview of plasma membrane microviscosity, and plasma membrane micro-organization, between the cells of the same line. Thus, we clearly reveal the higher heterogeneity of plasma membrane in multidrug-resistant cancer cells in comparison with the nonresistant ones (denoted sensitive cells). We also display distinct modifications related to a membrane fluidity modulator, benzyl alcohol, and two revertants of multidrug resistance, verapamil and cyclosporin-A. A relation between the distribution of the diffusion-time values and the modification of membrane lateral heterogeneities is proposed. PMID- 19566324 TI - Millimeter-resolution acousto-optic quantitative imaging in a tissue model system. AB - We have investigated the application of ultrasound modulated coherent light for quantitative determination of the ratio of dye concentrations and total concentration of absorbers in a blood vessel-mimicking sample. A 3-mm-diam tube containing the mixture of dyes inside an Intralipid-based gel with optical properties similar to tissue was interrogated by two different laser wavelengths in combination with intense microsecond ultrasound bursts. The use of calibration curves allowed us to extract quantitative information on the ratio of dye concentrations with the accuracy of better than 15%, as well as on the total concentration. Furthermore, we demonstrated the feasibility to obtain a quantitative 3-D map of the absorbing structure with a spatial resolution of better than 3 mm. These findings give an outlook to apply this technique for noninvasive 3-D mapping of oxygen saturation and total concentration of hemoglobin in tissue. PMID- 19566325 TI - Acoustic fingerprints of dye-labeled protein submicrosphere photoacoustic contrast agents. AB - Dye-labeled protein microspheres, submicron in size and capable of producing thermoelastically generated ultrasound in response to laser stimulation, are presented as contrast agents for photoacoustic imaging. Incident laser energy absorbed by fluorescein isothiocyanate (FITC)-labeled elastin submicrospheres results in thermoelastically generated sound production. Plotted A-line graphs reveal a distinctive morphology and a greater than two orders of magnitude increase in signal amplitude subsequent to converting FITC elastin into submicrospheres (despite a four orders of magnitude decrease in concentration). Evidence of nonlinearity and enhancement of ultrasound backscatter indicate a potential use in contrast-enhanced harmonic imaging. Photoacoustic and ultrasound imaging of FITC-elastin submicrospheres in a water-filled phantom vessel shows enhanced contrast at low concentration and clear delineation of the phantom vessel wall. PMID- 19566326 TI - Early diagnosis of melanotic melanoma based on laser-induced melanin fluorescence. AB - Because of the increasing incidence of skin cancer, interest in using the autofluorescence of skin tissue as a noninvasive tool for early diagnosis is enforced. Focus is especially on malignant melanotic melanoma. On the basis of a newly developed method to selectively excite melanin fluorescence of skin tissue by stepwise two-photon excitation with nanosecond laser pulses at 810 nm, we have investigated information from this melanin fluorescence with respect to the differentiation of pigmented lesions. A distinct difference in the melanin fluorescence spectrum of malignant melanoma (including melanoma in situ) when compared to that of benign melanocytic lesions (i.e., common nevi) has been found for freshly excised samples as well as for histopathological samples. There is also specific fluorescence from dysplastic nevi. In this way, early detection of malignant melanoma is possible. PMID- 19566327 TI - Automated identification of tumor microscopic morphology based on macroscopically measured scatter signatures. AB - An automated algorithm and methodology is presented to identify tumor-tissue morphologies based on broadband scatter data measured by raster scan imaging of the samples. A quasi-confocal reflectance imaging system was used to directly measure the tissue scatter reflectance in situ, and the spectrum was used to identify the scattering power, amplitude, and total wavelength-integrated intensity. Pancreatic tumor and normal samples were characterized using the instrument, and subtle changes in the scatter signal were encountered within regions of each sample. Discrimination between normal versus tumor tissue was readily performed using a K-nearest neighbor classifier algorithm. A similar approach worked for regions of tumor morphology when statistical preprocessing of the scattering parameters was included to create additional data features. This type of automated interpretation methodology can provide a tool for guiding surgical resection in areas where microscopy imaging cannot be realized efficiently by the surgeon. In addition, the results indicate important design changes for future systems. PMID- 19566328 TI - Multimodality bronchoscopic [corrected] imaging of tracheopathica osteochondroplastica. AB - Results of a commercial optical coherence tomography system used as part of a multimodality diagnostic bronchoscopy platform are presented for a 61-year-old patient with central airway obstruction from tracheopathica osteochondroplastica. Comparison to results of white-light bronchoscopy, histology, and endobronchial ultrasound examination are accompanied by a discussion of resolution, penetration depth, contrast, and field of view of these imaging modalities. White-light bronchoscopy revealed irregularly shaped, firm submucosal nodules along cartilaginous structures of the anterior and lateral walls of the trachea, sparing the muscular posterior membrane. Endobronchial ultrasound showed a hyperechoic density of 0.4 cm thickness. optical coherence tomography (OCT) was performed using a commercially available, compact time-domain OCT system (Niris System, Imalux Corp., Cleveland, Ohio) with a magnetically actuating probe (two dimensional, front imaging, and inside actuation). Images showed epithelium, upper submucosa, and osseous submucosal nodule layers corresponding with histopathology. To our knowledge, this is the first time these commercially available systems are used as part of a multimodality bronchoscopy platform to study diagnostic imaging of a benign disease causing central airway obstruction. Further studies are needed to optimize these systems for pulmonary applications and to determine how new-generation imaging modalities will be integrated into a multimodality bronchoscopy platform. PMID- 19566330 TI - Confocal reflectance and two-photon microscopy studies of a songbird skull for preparation of transcranial imaging. AB - We present experiments and analyses of confocal reflectance and two-photon microscopy studies of zebra finch skull samples. The thin and hollow structure of these birds' skulls is quite translucent, which can allow in vivo transcranial two-photon imaging for brain activation monitoring. However, the skull structure is also quite complex, with high refractive index changes on a macroscopic scale. These studies aim at exploring the geometrical and scattering properties of these skull samples with the use of several confocal microscopy contrasts. Moreover, the study of the axial reflectance exponential decay is used to estimate the scattering coefficients of the bone. Finally, two-photon imaging experiments of a fluorescent object located beneath the skull are carried out. It reveals that two photon fluorescence can be collected through the skull with a strong signal. It also reveals that the spatial resolution loss is quite high and cannot be fully explained by the bulk scattering properties of the bone, but also by the presence of the high refractive index inhomogeneity of this pneumatic skull structure. Even if the optical properties of the skull are different during in vivo experiments, these preliminary studies are aimed at preparing and optimizing transcranial brain activation monitoring experiments on songbirds. PMID- 19566329 TI - Mapping microscope object polarized emission to the back focal plane pattern. AB - The back focal plane (BFP) intensity pattern from a high-aperture objective separately maps far- and near-field emission from dipoles near a bare glass or metal-film-coated glass/aqueous interface. Total internal reflection (TIR) excitation of a fluorescent sample gave a BFP pattern interpreted in terms of fluorescent dipole orientation and distance from the interface. Theoretical consideration of this system led to identification of emission characteristics that remove a dipole orientation degeneracy in conventional microscope fluorescence polarization measurements. BFP pattern inspection removes the degeneracy. Alternatively, a BFP mask blocking a small fraction of emitted light in a standard imaging microscope prevents uniform collection of the BFP intensity and also eliminates the degeneracy. The BFP pattern from a single photoactivated photoactivatable green fluorescent protein (PAGFP) tagged myosin in a muscle fiber was observed despite the large background light from the highly concentrated myosin tagged with unphotoactivated PAGFP. This was accomplished by imaging the pattern from a nontelecentric plane, where most of the background intensity's pattern was translated laterally from the single-molecule object's pattern. TIR/BFP pattern imaging requires a simple alteration of the fluorescence microscope and is consistent with single-molecule imaging in a fluorophore dense three-dimensional object like a muscle fiber. PMID- 19566331 TI - Enhanced yellow fluorescent protein photoconversion to a cyan fluorescent protein like species is sensitive to thermal and diffusion conditions. AB - Ongoing research efforts into fluorescent proteins continuously generates new mutation variants, some of which can become photoactivated or photoconverted to a red-shifted color upon intense UV or blue light illumination. We report a built in propensity for enhanced yellow fluorescent protein (EYFP) to undergo irreversible photoconversion into a cyan fluorescent protein (CFP)-like species upon green-light illumination. The photoconversion is thermally activated, happens mainly in fixed, nonsealed cell samples, and may result in a very bright and relatively photostable CFP-like species. The photoconversion efficiency depends on the sample diffusivity and is much increased in dehydrated, oxygenated samples. Given the large variations in conversion efficiency observed among samples as well as within a sample, photoconversion cannot be appropriately accounted for in the analysis of acceptor photobleaching fluorescence resonance energy transfer (pbFRET) images and should rather be completely avoided. Thus, samples should always be checked and discarded if photoconversion is observed. PMID- 19566333 TI - Selection of optimal wavelengths for spectral reconstruction in diffuse optical tomography. AB - Using lasers with different wavelengths in diffuse optical tomography (spectral DOT) has the advantage that the concentrations of chromophores can be reconstructed quantitatively. In continuous wave spectral DOT, it is furthermore possible to distinguish between scattering and absorption. The choice of the laser wavelengths has a strong impact on how well the scattering parameter and chromophore concentrations can be determined. Current methods to optimize the set of wavelengths disregard the sensitivity of the reconstruction result to uncertainties in the absorption spectra of the chromophores. But since available absorption spectra show significant deviations, it seems to be necessary to take this into account. The wavelength optimization approach presented here is an extension to a method of Corlu et al. The original method optimizes the wavelength sets such that scattering parameters and chromophore concentrations can be separated optimally. We introduce an additional criterion that evaluates the dependence of reconstructed chromophore concentrations on deviations of the extinction coefficients. The wavelength sets found by the new approach are different from those determined with the original method. Reconstructions of simulated data show the effect of using various absorption spectra for reconstruction with different wavelength sets and illustrate the advantages of the new wavelength sets. PMID- 19566332 TI - Automated algorithm for breast tissue differentiation in optical coherence tomography. AB - An automated algorithm for differentiating breast tissue types based on optical coherence tomography (OCT) data is presented. Eight parameters are derived from the OCT reflectivity profiles and their means and covariance matrices are calculated for each tissue type from a training set (48 samples) selected based on histological examination. A quadratic discrimination score is then used to assess the samples from a validation set. The algorithm results for a set of 89 breast tissue samples were correlated with the histological findings, yielding specificity and sensitivity of 0.88. If further perfected to work in real time and yield even higher sensitivity and specificity, this algorithm would be a valuable tool for biopsy guidance and could significantly increase procedure reliability by reducing both the number of nondiagnostic aspirates and the number of false negatives. PMID- 19566334 TI - Assessment of the effects of ultrasound-mediated alcohols on skin optical clearing. AB - Our previous studies have shown the ultrasound-induced skin optical clearing enhancement with topical application of glycerol on in vitro porcine skin and in vivo human skin. The objective of this study was to find more effective ultrasound-alcohol combinations on skin optical clearing. The effect of sonophoretic delivery (SP) in combination with a series of alcohols such as glycerol, propylene glycol, butanediol, butanol, polyethylene glycol (PEG200, PEG400) on skin optical clearing was investigated. Light transmittance and imaging depth of in vitro porcine skin were measured with spectroscopy and optical coherence tomography (OCT). Sixty percent alcohols and ultrasound (SP) with a frequency of 1 MHz and a power of 0.75 W over a 3-cm probe was simultaneously applied for 15 min. Optical clearing capability of alcohols was much improved with the simultaneous application of ultrasound compared with alcohols alone. Light transmittance at 1276 nm increased by 31% (butanol) to 57% (PEG200) at 60 min, respectively. PEG200 with ultrasound exhibited 1.8-fold the clearing capability of PEG200 alone, which was the highest enhancement by ultrasound among the six alcohols. Sixty minutes of PEG200/SP exposure resulted in a significantly increased 1e light penetration depth of 1.43 mm compared to 0.95 mm for control in OCT measurement. PMID- 19566335 TI - New method for the detection of micro-organisms in blood: application of quantitative interpretation model to aerobic blood cultures. AB - The physical and chemical changes occurring in blood that has been inoculated into a blood culture bottle can be used as means to detect the presence of microorganisms in blood cultures. These changes include primarily the conversion of oxy- to deoxyhemoglobin within the red blood cells (RBCs) and changes in the cell number densities. These changes in the physical and chemical properties of blood can be readily detected using spectrophometric methods thus enabling the continuous monitoring of blood culture vials to provide quantitative information on the growth behavior of the microorganisms present. This paper reports on the application of spectrophotometric information obtained from diffuse reflectance measurements of aerobic blood cultures to detect microbial growth and compares the results to those obtained using the standard blood culture system. PMID- 19566336 TI - Selection of regularization parameter for optical topography. AB - The choice of the regularization parameter has a profound effect on the solution of ill-posed inverse problems such as optical topography. We review 11 different methods for selecting the Tikhonov regularization parameter that have been described previously in the literature. We test them on two trial problems, deblurring and optical topography, and conclude that the L-curve method is the method of choice, though in particularly ill-posed problems, generalized cross validation may provide an alternative. PMID- 19566337 TI - Three-dimensional surface profile intensity correction for spatially modulated imaging. AB - We describe a noncontact profile correction technique for quantitative, wide field optical measurement of tissue absorption (microa) and reduced scattering (micros) coefficients, based on geometric correction of the sample's Lambertian (diffuse) reflectance intensity. Because the projection of structured light onto an object is the basis for both phase-shifting profilometry and modulated imaging, we were able to develop a single instrument capable of performing both techniques. In so doing, the surface of the three-dimensional object could be acquired and used to extract the object's optical properties. The optical properties of flat polydimethylsiloxane (silicone) phantoms with homogenous tissue-like optical properties were extracted, with and without profilometry correction, after vertical translation and tilting of the phantoms at various angles. Objects having a complex shape, including a hemispheric silicone phantom and human fingers, were acquired and similarly processed, with vascular constriction of a finger being readily detectable through changes in its optical properties. Using profilometry correction, the accuracy of extracted absorption and reduced scattering coefficients improved from two- to ten-fold for surfaces having height variations as much as 3 cm and tilt angles as high as 40 deg. These data lay the foundation for employing structured light for quantitative imaging during surgery. PMID- 19566338 TI - Automated segmentation of tissue structures in optical coherence tomography data. AB - Segmentation of optical coherence tomography (OCT) images provides useful information, especially in medical imaging applications. Because OCT images are subject to speckle noise, the identification of structures is complicated. Addressing this issue, two methods for the automated segmentation of arbitrary structures in OCT images are proposed. The methods perform a seeded region growing, applying a model-based analysis of OCT A-scans for the seed's acquisition. The segmentation therefore avoids any user-intervention dependency. The first region-growing algorithm uses an adaptive neighborhood homogeneity criterion based on a model of an OCT intensity course in tissue and a model of speckle noise corruption. It can be applied to an unfiltered OCT image. The second performs region growing on a filtered OCT image applying the local median as a measure for homogeneity in the region. Performance is compared through the quantitative evaluation of artificial data, showing the capabilities of both in terms of structures detected and leakage. The proposed methods were tested on real OCT data in different scenarios and showed promising results for their application in OCT imaging. PMID- 19566339 TI - Photophysical properties of Cerulean and Venus fluorescent proteins. AB - Cerulean and Venus are recently developed fluorescent proteins, often used as a donor-acceptor pair by researchers in Forster resonance energy transfer-based colocalization studies. We characterized the fluorescent properties of these two proteins in a broad spectral range (form ultraviolet to visible region). Excitation spectra, lifetimes, and polarization spectra show significant energy transfer from aromatic amino acids to the fluorescent protein chromophore. High steady-state anisotropy values and the lack of a fast component in anisotropy decays show that the fluorescent protein chromophore is rigidly fixed within the protein structure. Furthermore, we show that the chromophores are not accessible to external quenchers, such as acrylamide or potassium iodide (KI), allowing the removal of "unwanted" background in the environment with external quencher, while leaving the Cerulean/Venus fluorescence unchanged. PMID- 19566340 TI - Multiphoton adaptation of a commercial low-cost confocal microscope for live tissue imaging. AB - The Nikon C1 confocal laser scanning microscope is a relatively inexpensive and user-friendly instrument. We describe a straightforward method to convert the C1 for multiphoton microscopy utilizing direct coupling of a femtosecond near infrared laser into the scan head and fiber optic transmission of emission light to the three-channel detector box. Our adapted system can be rapidly switched between confocal and multiphoton mode, requires no modification to the original system, and uses only a few custom-made parts. The entire system, including scan mirrors and detector box, remain under the control of the user-friendly Nikon EZ C1 software without modification. PMID- 19566341 TI - Noninvasive characterization of the fission yeast cell cycle by monitoring dry mass with digital holographic microscopy. AB - Digital holography microscopy (DHM) is an optical technique which provides phase images yielding quantitative information about cell structure and cellular dynamics. Furthermore, the quantitative phase images allow the derivation of other parameters, including dry mass production, density, and spatial distribution. We have applied DHM to study the dry mass production rate and the dry mass surface density in wild-type and mutant fission yeast cells. Our study demonstrates the applicability of DHM as a tool for label-free quantitative analysis of the cell cycle and opens the possibility for its use in high throughput screening. PMID- 19566342 TI - Feasibility of digitally stained multimodal confocal mosaics to simulate histopathology. AB - Fluorescence confocal mosaicing microscopy of tissue biopsies stained with acridine orange has been shown to accurately identify tumors and with an overall sensitivity of 96.6% and specificity of 89.2%. However, fluorescence shows only nuclear detail similar to hematoxylin in histopathology and does not show collagen or cytoplasm, which may provide necessary negative contrast information similar to eosin used in histopathology. Reflectance mode contrast is sensitive to collagen and cytoplasm without staining. To further improve sensitivity and specificity, digitally stained confocal mosaics combine confocal fluorescence and reflectance images in a multimodal pseudo-color image to mimic the appearance of histopathology with hematoxylin and eosin and facilitate the introduction of confocal microscopy into the clinical realm. PMID- 19566345 TI - Plagiorchis elegans (Trematoda) and incompatible snail hosts: implications for snail life history traits and biocontrol of human schistosomiasis. AB - We examined the effect of Plagiorchis elegans on egg production and survival on Bulinus truncatus and Helisoma trivolvis trivolvis. Both species are incompatible hosts for P. elegans. Helisoma t. trivolvis occurs sympatrically with P. elegans; B. truncatus does not. Overall, P. elegans had no effect on survivorship or egg production in H. t. trivolvis or on the survivorship of B. truncatus. Its effect on egg production in B. truncatus was transitory; egg production was reduced by 50% for 5 wk following exposure but returned to normal thereafter. Neither egg production nor survivorship was affected in adult H. t. trivolvis. Egg production ceased at 14 wk post-exposure (PE), but resumed when the snails were paired. Young H. t. trivolvis also produced eggs after exposure, but later than the adults and only after they had been paired with another snail. This suggests that a need for periodic cross-fertilization in H. t. trivolvis rather than the effect of the parasite is responsible for the cessation of egg production in this species. Survivorship in young H. t. trivolvis was significantly higher in exposed snails between wk 7 to 10 PE than in controls. PMID- 19566346 TI - A new metastrongilid species (Nematoda: Metastrongylidae): a lungworm from Akodon montensis (Rodentia: Sigmodontinae) in Brazil. AB - Angiostrongylus lenzii n. sp. (Nematoda: Metastrongyloidea) is described and illustrated from the pulmonary artery of the wild rodent Akodon montensis Thomas, 1913, collected in the municipality of Teresopolis, Rio de Janeiro State, Brazil. It represents the 17th species of the genus and the 2nd in South America as parasites in rodents. This new species is the most similar morphologically to Angiostrongylus vasorum, Angiostrongylus gubernaculatus, Angiostrongylus schmidti, and Angiostrongylus morerai. It can be distinguished from them by the morphology of the caudal bursa, ventral rays 2 and 3 with conspicuous knobs, differences in width and length of lateral and dorsal rays, presence of rays 8, and length of spicules. PMID- 19566347 TI - New species of rhynchobdellid leech (Hirudinea: Glossiphoniidae): a parasite of turtles from Chiapas, Mexico. AB - A new species of rhynchobdellid leech, Placobdella ringueleti n. sp., is described based on the examination of 25 specimens. Leeches were collected from body appendages of freshwater turtles (Kinosternon leucostomum, Dermatemys mawii, and Staurotypus triporcatus ) from Chiapas, Mexico. The new species resembles other members of the genus in the ocular morphology, bilobated ovaries, and presence of 1 pair of elongated mycetomes, but it is distinguished from them by the dorsal papillar pattern, by having a continuous longitudinal mid-dorsal stripe along the dorsal surface and papillae on the dorsal surface of the posterior sucker. Placobdella ringueleti represents the third species of the genus in Mexico. The type locality of the new species is part of the "El Ocote" Biosphere Reserve, Chiapas, Mexico. PMID- 19566348 TI - The Iranian connection. PMID- 19566349 TI - An estimation of the chronic rejection of kidney transplant using an eternal Weibull regression: a historical cohort study. AB - BACKGROUND: We estimated the chronic rejection of kidney transplant using an eternal Weibull regression. METHODS: In this historical cohort study, we enrolled all patients with chronic renal failure who were admitted to Shahid Labbafinejad medical center (Tehran, Iran) from 1984 to 2003. Using Matlab 7.0, we considered the eternal proportion theta, as a logistic-type function of the covariates and modified the survival function. We estimated the survival function in unmodified and modified forms using Weibull distribution. RESULTS: The chance of chronic rejection was 1.95 times higher among those who received a kidney transplant before 1996. Considering all cases who received renal transplantation after 1984, males had a chance of rejection 20% less than females. Next to the eternity, Weibull model was fitted to patients who received renal transplantation after 1996. Treatment protocol was changed after 1996 expecting fewer chronic rejections; thereafter, the eternal proportion was estimated to be 0.81. This seems quite considerable as a percentage of non-failure cases. CONCLUSION: Providing a non-zero eternal proportion, the modified model would be superior over the unmodified model. PMID- 19566350 TI - Practical strategies to reduce morbidity and mortality of natural catastrophes: a retrospective study based on Bam earthquake experience. AB - BACKGROUND: Critical analysis of shortcomings of emergency medical management of earthquake casualties will provide an invaluable insight to improve outcomes for future events. Using a critical analysis methodology to evaluate the quality of emergency medical management after Bam earthquake, we suggest a practical strategic approach to decrease morbidity and mortality after such events. METHODS: We designed a questioner to register the basic demographic data and the key biologic parameters of all rescued victims arriving in hospitals. Based on that questioner a data bank was created and used for different analyses. In addition, published official reports and on the scene observations of our nephrologist colleagues were other sources of our data. RESULTS: Bam earthquake was publicly announced more than six hours after its occurrence. The earliest time when local and international rescue teams arrived at the scene was 12 hours after the disaster. Fifty-four percent of hospital inpatients had been admitted on the second or third day after the earthquake. The mean time of being under the rubble was 4.8+/-4.9 hours. The mean time between extrication and initiation of intravenous fluid infusion was 18.9 hours (min: 10 minutes, max: 96 hours). CONCLUSION: Problems encountered in the aftermath of the Bam earthquake were related to the lack of prepared action plan and data management system. Here, we present a specifically designed earthquake chart. By following the chart, rescue paramedic personnel and emergency medical teams will be able to recognize high risk victims, in order to provide timely medical management. PMID- 19566351 TI - The role of endoscopic autopsy in trauma cases: the first report from Iran. AB - BACKGROUND: The traditional autopsy is unpleasant for the family members of the dead person. The aim of this study was to assess abdominal traumas by a laparoscope and to determine the accuracy of laparoscopic examination in comparison with the traditional autopsy. METHODS: From December 2004 through August 2005, 50 fresh cadavers (<24 hours old) of blunt trauma victims were selected. They were first assessed by a laparoscope for intraperitoneal and retroperitoneal organs and then the traditional autopsy was performed as gold standard. The organs were assessed regarding the nature and intensity of damages and the results were compared with laparoscopic findings. The diagnostic accuracy of laparoscope was determined for each case with 95% confidence intervals (CIs) using Fisher's exact test. RESULTS: The accuracy for investigation of intraperitoneal and retroperitoneal organs and the overall accuracy values were significant. The accuracy of laparoscope for detecting intraperitoneal organ damage was 90% (95%CI of 81.7%to 94.8%) while for the retroperitoneal organs was 92% (95%CI of 84.7% to 96%). The overall accuracy of laparoscope was 84% (with 95%CI of 74.3% to 90.5%). CONCLUSION: The sensitivity and specificity of laparoscopic examination for intraperitoneal organs are acceptable but its accuracy for retroperitoneal organs is lower. Therefore, it seems that laparoscopic examination is an eligible substitute for the traditional autopsy. PMID- 19566352 TI - The prevalence of cigarette smoking in residents of Tehran. AB - BACKGROUND: To determine the prevalence of cigarette smoking in Tehran and some of its determinants. METHODS: In a cross-sectional population-based study, a sample from citizens of Tehran was obtained using stratified random cluster sampling. Through a face to face interview, data such as the history of smoking in the past (abstinence from smoking) and at the present time (active smoking), and number of cigarettes per day and per year were collected. In general, people who smoked more than 100 cigarette-days were considered smokers. Pack per year was used to evaluate the severity of smoking. RESULTS: Out of the sampled people, 4565 participated in the study. Of these, 74.4%, who were over 15 years of age, were studied for smoking. The prevalence of smoking in Tehran was 11.9% (95%CI, 10.6-13.3). Some 20.6% of the male participants (95%CI, 18.2-23.1) and 2.9% of the female participants (95%CI, 2.2-3.7) were smokers (P<0.001). The prevalence of smoking significantly increased with age until 54 years of age (OR=1.07; 95%CI, 1.06-1.08). People with higher education smoked more. Also, married people smoked significantly more than singles (OR=2.7; 95%CI, 1.85-3.95). Smokers used 209 cigarette pack-years on average, which was significantly higher in men and in people with lower education (P<0.001). CONCLUSION: The results of this study showed lower prevalence of cigarette smoking has relatively decreased compared with previous studies. However, healthcare officials should consider educational and healthcare programs to decrease the prevalence of smoking. PMID- 19566353 TI - Quality of life improvement in hearing-impaired elderly people after wearing a hearing aid. AB - BACKGROUND: Communication is an important aspect of everyday life especially for elderly people. Hearing loss can impair the exchange of information and therefore reduces the quality of life. The purpose of this study was to investigate the quality of life in elderly people who are hard of hearing after wearing a hearing aid. METHODS: Hearing-impaired old people who referred to the rehabilitation centers of Tehran Welfare Organization for having a hearing aid completed the Hearing Handicap Inventory for the Elderly questionnaire before and three months after using a hearing aid. Scores of the effects of hearing loss on quality of life were compared for each subject and between males and females. RESULTS: The results showed a significant improvement of the quality of life after three months of using a hearing aid in all participants and betterment of their most important problems i.e., the communication and exchange of information. CONCLUSION: In conclusion, with respect to the beneficial effects of hearing aids in presbycusis and due to the significant improvement in the quality of life, hearing aids are recommended for this group of population. PMID- 19566354 TI - Immunophenotypic pattern and cytokine profiles of dry type cutaneous leishmaniasis. AB - BACKGROUND: Dry type localized cutaneous leishmaniasis, one of the most prevalent cutaneous parasitic infections in Kerman Province, is presented as a polarized disease in which cytokine profiles secreted by immune cells play a major role in its presentation. In order to clarify the idea, immunohistochemical study of skin biopsies were performed to elucidate the cytokine release capabilities of immune cells. METHODS: Skin biopsies of acute, chronic nonlupoid, and chronic lupoid recidivans lesions of dry type localized cutaneous leishmaniasis were studied by immunohistochemical staining methods for immunophenotypic patterns (CD4, CD8, CD14, CD19, CD56, CD11a, CD18, CD1a, HLA-DR, CD54) and cytokines (INF-gamma, IL 12, IL-4, TNF-alpha) released by immune inflammatory cells. RESULTS: The descriptive analysis of data showed that the mean percentage of positive immunostained cells of CD4, CD8, and CD14; antigen-presenting cells (CD1a, HLA DR); and markers of the extravasated positive memory T cells (CD11a, CD18, CD54) are more frequent in lupoid recidivans than in acute active and chronic nonlupoid lesions, in order of frequency. CONCLUSION: Based on the results, it seems that Th1-like response is predominant in acute active form and lupoid recidivans while Th2-like response is predominant in chronic nonlupoid lesions. It seems that lupoid recidivans is a type IV hypersensitivity reaction to the reactivation of hidden antigens. PMID- 19566355 TI - Determining the inter- and intraobserver reproducibility of the diagnosis of endometrial hyperplasia subgroups and well-differentiated endometrioid carcinoma in endometrial curettage specimens. AB - BACKGROUND: Many studies have attempted to identify histologic features that help to distinguish atypical hyperplasia from hyperplasia without atypia and well differentiated endometrioid carcinoma of the endometrium; however, few have evaluated the reproducibility of these diagnoses. METHODS: Five pathologists independently reviewed 100 endometrial curettage specimens chosen to represent the spectrum of proliferative lesions of the endometrium. This included simple hyperplasia, complex hyperplasia, atypical hyperplasia, and well-differentiated endometrioid carcinoma. Slides were reviewed once for interobserver agreement among the five pathologists and twice for intraobserver agreement by one of them. RESULTS: The results were assessed using the weighted kappa statistic. The mean intraobserver kappa value was 0.86. The mean interobserver kappa values by diagnostic category were as follows: simple hyperplasia without atypia: 0.74; complex hyperplasia without atypia: 0.33; atypical hyperplasia: 0.34, and well differentiated endometrioid carcinoma: 0.64; with a kappa value of 0.53 for all cases combined. CONCLUSION: A major interobserver discrepancy exists in the diagnosis of complex and atypical hyperplasia which are the most similar mimics of endometrioid carcinoma. PMID- 19566356 TI - Ultrasonographic screening for abdominal aortic aneurysms in Iranian candidates for coronary artery bypass graft surgery. AB - BACKGROUND: To evaluate the prevalence of abdominal aortic aneurysm in Iranian candidates for coronary artery bypass graft surgery and its associated factors. METHODS: Totally, 2843 consecutive candidates for coronary artery bypass graft surgery underwent ultrasonography of abdominal aorta. The relation between abdominal aortic aneurysm with gender, age, smoking, dyslipidemia, hypertension, diabetes, and carotid and coronary stenosis was evaluated. RESULTS: The prevalence of abdominal aortic aneurysm was 2.7% and 0.7% in men and women subgroups, respectively. The prevalence was 3.8% in men older than 65 years. The largest diameter of abdominal aortic aneurysm was 61.7 mm. Only two men had aneurysm larger than 55 mm, which led to changing their surgery schedule to stenting. Gender, age, smoking, smoking more than 40 years, diabetes , hypertension, and significant carotid stenosis were associated factors of abdominal aortic aneurysm. CONCLUSION: Compared with the results sited in Western studies, the prevalence of abdominal aortic aneurysm is lower in Iran (P<0.0001). Its associated factors in our study were similar to previously published studies. Significant carotid stenosis, a factor not reported previously, was an associated factor in our study. Because of the low prevalence of abdominal aortic aneurysm in our population, screening the candidates for coronary artery bypass graft surgery is not recommended. PMID- 19566357 TI - Totally implantable subpectoral vs. subcutaneous port systems in children with malignant diseases. AB - BACKGROUND: For many years, subcutaneous therapeutic port system was known as a major route to access central veins. However, significant complications have been reported through recent years. One of the most important complications of subcutaneous port implantation is skin necrosis. In order to decrease this complication, we would like to introduce subpectoral fascia port implantation through this study. METHODS: Five hundred and twenty four patients with a variety of neoplastic diseases underwent port implantation, from March 2003 to March 2008 (60 months). All suitable size catheters were put in the superior vena cava through the internal jugular vein under general anesthesia. The ports were placed in the subcutaneous pocket (SCP group) in 342 patients and in the subpectoral fascia pocket (SPFP group) in 182 patients. Data were analyzed using Chi-square test and survival analysis for time (Kaplan-Meier). RESULTS: A total of 538 devices were placed for 524 patients in two groups (14 patients received a second device after removal of the first one, due to failure of the first implantation). Mean follow-up period was 508 days (8 - 2025 days).Common complications observed in the SPFP group were as follows: wound infection (7 cases, 3.8%), catheter obstruction (7 cases, 3.8%), catheter displacement (6 cases, 3.2%), port related infection (5 cases, 2.7%), and pocket hematoma (2 cases, 1.1%).Common complications observed in the SCP group were as follows: catheter displacement (12 cases, 3.5%), skin necrosis (11 cases, 3.21%), port exposure (9 cases, 2.6%), port related infection (8 cases, 2.3%), catheter obstruction (8 cases, 2.3%), and port rotation (3 cases, 0.9%). CONCLUSION: The results showed that port implantation in the subpectoral fascia pocket had a lower rate of skin complications than the subcutaneous pocket implantation. According to this study, this procedure was not complicated by skin necrosis over the port, port exposure or port rotation. PMID- 19566358 TI - Cancer-testis antigens: potential targets for cancer immunotherapy. AB - Cancer-testis antigens are tumor antigens that their expression is almost limited to male germ cells in the testis. Some of cancer-testis antigens are also expressed in the ovary and in trophoblasts. Recently their expression has been seen in different types of tumors. Many pathophysiologic studies suggest that a blood-testis barrier exists in the testis. Because spermatogenesis begins at puberty, new cell-surface antigens are expressed when the immune system has refined the ability to distinguish self from nonself. So, sperms in the testis do not stimulate immune responses. In addition, although antigen-presenting cells are commonly seen in the interstitial spaces of the testis, these cells are scarcely seen within the seminiferous tubules. So, testis is considered as an immune-privileged site, and testis-specific genes, if expressed in cancers can be immunogenic. For this reason cancer-testis antigens are promising candidates for cancer immunotherapy and have become a major focus for the development of vaccine based clinical trials in recent years. In addition, these antigens can also be used as biomarkers for early detection of cancers. PMID- 19566359 TI - An overview of heel Marjolin's ulcers in the Orthopedic Department of Urmia University of Medical Sciences. AB - Marjolin's ulcer is defined as a malignant, ulcerating neoplasm occurring in cicatricial tissues. The cancer is usually a well-differentiated squamous cell carcinoma. Wide resection is complicated with a recurrence rate of 20% to 50% and a metastasis rate of 54%. Therefore, we chose amputation as the modality of treatment for heel Marjolin's ulcers in Urmia and presented their results in this study.During the last 10 years in Urmia, nineteen cases of heel Marjolin's ulcers has been detected. About 47% were due to childhood burn. Malignancy was mainly squamous cell carcinoma. The mean latent period of malignant transformation was 11 years. All cases were treated with amputation, without any recurrence or metastasis in an average four-year follow-up period.The squamous cell carcinoma of Marjolin's ulcer has the worst prognosis in comparison with other squamous cell carcinomas and it requires an aggressive treatment. PMID- 19566360 TI - Surgical intervention for treatment of septic arthritis in infancy and childhood; a retrospective study. AB - Septic arthritis is a fearful condition because of its mortality and the potentially late sequels on immature skeleton including limb shortening, osteomyelitis, destruction of joint surface, severe limitation of motion, and dislocation.This study was performed to reveal the final outcome of our patients and find out the possible risk factors of poor result. The case records of 243 children who were admitted with the diagnosis of septic arthritis in Imam Khomeini and Bu Ali Sina Hospitals, Mazandaran Province, were studied between 1996 and 2005. The diagnosis was based on clinical and ultrasound findings in all patients and positive smear in 67% of them. Among these patients, we had access to 162 cases who had definitely septic arthritis and went through surgical interventions because of the involvement of the hip joint or uncertain response to medical treatments. As four out of six poor outcome cases were related to hip sepsis, hip was the main site of involvement and complications.Six cases of severe complications out of 162 showed the favorite result due to early diagnosis and intervention and highlighted the grave prognostic factors which were delayed diagnosis, infantile age and hip sepsis. PMID- 19566361 TI - Bilateral invasive ductal carcinoma in a woman with neurofibromatosis type 1. AB - A 51-year-old woman with neurofibromatosis type 1 presented to our department for investigation of a left breast lump (50 x 50 x 30 mm); a mass in the right breast (40 x 40 x 20 mm) was also detected on physical examination. The lumps were suspected to be malignant based on physical examination and ultrasonography. Biopsy and frozen sections subsequently confirmed a diagnosis of bilateral invasive ductal carcinoma. A standard bilateral radical mastectomy was performed, followed by postoperative chemoendocrine therapy. Tumor recurrence has not been observed within the first 23 months following the surgery. PMID- 19566362 TI - Palmaris profundus as the cause of carpal tunnel syndrome. AB - This report presents a palmaris profundus tendon anomaly in the carpal tunnel. The case was a 19-year-old female who underwent carpal tunnel surgery. After dividing the transverse carpal ligament, the palmaris profundus tendon was discovered. The abnormal tendon was within the carpal tunnel, laying over the investing tissue of the median nerve, and inserted deeply into the palmar aponeurosis. A segment of the abnormal tendon was excised and the patient's symptoms resolved completely. PMID- 19566363 TI - Ameloblastomatous calcifying odontogenic cyst: a case report of a rare histologic variant. AB - Calcifying odontogenic cyst is an uncommon developmental odontogenic cyst first described by Gorlin in 1962. It is considered as an extremely rare cyst and accounts for only 1% of jaw cysts reported. Because of its diverse histopathology, there has always been confusion about its nature as a cyst, neoplasm, or hamartoma. In this report, we present a rare case of calcifying odontogenic cyst with ameloblastic proliferation--an extremely rare histologic variant in a 22-year-old male in the right mandibular molar region. The lesion was surgically removed. After enucleation no recurrence has been recorded in the ensuing 14 months. PMID- 19566364 TI - Maffucci's syndrome with oral manifestations. AB - Maffucci's syndrome is a rare congenital mesodermal dysplasia combined with dyschondroplasia and hemangiomatosis. About 150 cases have been reported till now. Maffucci's syndrome is often combined with other neoplasms. Herein, we report a patient with Maffucci's syndrome and hemangiomas on the dorsum of the tongue, which is rare in this syndrome. PMID- 19566365 TI - Esophageal duplication cyst. PMID- 19566366 TI - Development of gastroenterology and hepatology in Iran: part I-training programs and the Iranian association of gastroenterology and hepatology. AB - During the first half of the 20th century, all subspecialties of internal medicine were managed by general internists and there was no gastroenterologist in Iran.1 The first group of Iranian gastroenterologists who had completed their training program in France started working in Tehran in 1951. Later on, other specialists trained in USA and the United Kingdom joined them. Some of these specialists and some internists with special interest in gastroenterology and hepatology were in charge of gastroenterology education in divisions of internal medicine in the Medical Schools of Tehran, Shiraz, Isfahan, and other universities, while most of them were also practicing gastroenterology in their private offices on a part-time basis. The establishment of subspecialty training program in gastroenterology and hepatology was firstly started in 1976 in Tehran University and later on and since 1984 in Shaheed Beheshti, and Shiraz Universities. This was a major step in the progress of this subspecialty, which is now one of the most active medical disciplines in Iran.2 In this paper, we aimed to describe the development and evolution of gastroenterology and hepatology in Iran during the last 60 years with emphasis on physicians and scientists, who played a major role in providing the first gastroenterology services in the country or had a major role in training, education, and research in this field. We also describe the attempts that led to the establishment of the Iranian Association of Gastroenterology and Hepatology, as the most important nongovernmental organization in this field. PMID- 19566367 TI - "Ibn Sina and the clinical trial". PMID- 19566369 TI - Re: The prevalence of hepatitis B surface antigen and anti-hepatitis B core antibody in Iran: a population-based study. PMID- 19566370 TI - Oral breathing and dental malocclusions. AB - AIM: Aim of the present study was to evaluate existing correlations between oral breathing and dental malocclusions. METHODS: The study was conducted on a paediatric group of 71 oral breathers selected at the Allergology and Paediatric Immunology Department of Umberto I General Hospital, University of Rome "La Sapienza" (Italy). The children were selected based on inclusion/exclusion criteria. Children aged 6 to 12 years with no history of craniofacial malformations or orthodontic treatment were included. The results were compared with a control group composed of 71 patient aged 6 to 12 years with nasal breathing. After their medical history was recorded, all patients underwent orthodontic/otolaryngological clinical examinations. The following diagnostic procedures were then performed: latero-lateral projection teleradiography, orthopantomogram, dental impressions, anterior rhinomanometry before and after administering a local vasoconstrictor, nocturnal home pulse oximetry (NHPO) recording, spirometry test, skin prick test, study cast evaluation and cephalometric analysis following Tweed's principles. The intraoral examination assessed: dental class type, overbite, overjet, midlines, crossbite, and presence of parafunctional oral habits such as atypical swallowing, labial incompetence, finger sucking and sucking of the inner lip. Evaluation of the study casts involved arch perimeter and transpalatal width assessment, and space analysis. RESULTS: The results showed a strong correlation between oral breathing and malocclusions, which manifests itself with both dentoskeletal and functional alterations, leading to a dysfunctional malocclusive pattern. CONCLUSION: According to the authors' results, dysfunctional malocclusive pattern makes it clear that the association between oral breathing and dental malocclusions represents a self-perpetuating vicious circle in which it is difficult to establish if the primary alteration is respiratory or maxillofacial. Regardless, the problem needs to be addressed and solved through the close interaction of the paediatrician, otorhinolaryngologist, allergologist and orthodontist. PMID- 19566371 TI - Dental and periodontal health status in children affected by cystic fibrosis in a southern Italian region. AB - AIM: The aim of this study was to record the oral health status through the mean value of the DMFT/dmft index, the CPITN values and the DDE modified index in children affected by cystic fibrosis (CF). STUDY DESIGN: Observational study. METHODS: A group of 54 CF patients, aged 7 to 12 years, was selected and their data were compared to those of the same age healthy control group of 101 children. RESULTS AND STATISTICS: CF patients showed a mean DMFT (1.5 +/- 2.17) and dmft (0.42 +/- 0.9) significantly lower than control subjects (respectively 3.70 +/- 3.92 and 2.96 +/- 3.30). Moreover, in the CF subjects a high prevalence (55.6%) of enamel defects and a better periodontal health were found, despite the low presence of calculus in both groups. CONCLUSION: The low caries experience and the high prevalence of enamel defects in CF patients could be due to the metabolic disease, and above all to the long-term pharmacological therapies (antibiotics and pancreatic enzymes) that they take. PMID- 19566372 TI - Orthodontic treatment needs in an urban Iranian population, an epidemiological study of 11-14 year old children. AB - OBJECTIVES: A preliminary study to investigate the orthodontic treatment need and its gender distribution in urban Iranian schoolchildren. STUDY DESIGN: A cross sectional study was carried out at the school premises. SUBJECTS AND METHODS: For this survey, 502 subjects attending 6 different schools in the city of Isfahan, Iran (253 females and 249 males, aged 11-14 years) were randomly selected and examined, including those who were wearing an orthodontic appliance at the time of the survey (1 female and 5 males). IOTN (DHC and AC) scores were recorded for those who were not undergoing orthodontic treatment. One examiner, who had been formally calibrated in the use of occlusal indices, screened all schoolchildren. IOTN scores was calculated from direct examination. Gender dimorphism was evaluated by the Chi-square test (AC and DHC components of IOTN index). The prevalence of most severe occlusal traits in those who scored DHC 4 and 5 was calculated. RESULTS: At the time of the survey, 1.1% of subjects were wearing an orthodontic appliance, 36.1% had definite need (DHC 4 or 5) for orthodontic treatment, 20.2% borderline need (DHC 3) and 43.8% showed slight or no need for treatment (DHC 1 or 2). Reviewing the AC scores, 17.9% of the studied subjects showed definite need (AC 8-10) for orthodontic treatment, 36.1% borderline need (AC 5-7) and 46% showed slight or no need for treatment (AC 1-4). The prevalence of the four most severe occlusal traits in those with definite orthodontic treatment need were: severe maxillary crowding (43.6%), increased overbite (39.1%), increased overjet (35.8%) and severe mandibular crowding (27.4%). No gender differences were found for AC (P>0.05) and DHC (P>0.05) of IOTN index. CONCLUSION: Approximately one-third of Iranian school children were in need of orthodontic treatment, as determined by the index of orthodontic treatment need. PMID- 19566373 TI - Enamel and dentine of deciduous teeth Er:YAG laser prepared. A SEM study. AB - AIM: The aim of this study was to observe dentine and enamel surfaces of deciduous teeth under SEM after cavity preparation with Er:YAG laser using different fluences. The results showed that when using Er:YAG laser for cavity preparation in deciduous teeth, no carbonisation or cracks were observed on the enamel and dentine surfaces using energy output between 150-250 Mj, and frequency 15 Hz. The SEM images of the dentine and enamel surfaces were similar to previous studies on permanent teeth: enamel with a typical "lava flow" appearance as a result of an open core of the prism that has lost its typical hexagonal aspect and the dentine with opened tubules plus a difference in the mineral thickness between peritubular and intertubular. CONCLUSION: The difference between the SEM images of deciduous enamel and dentine when using three different energies (150 200-250 mJ) is not significant in order to recommend the use of one type of output energy. In addition, the SEM images are similar TO those of permanent teeth. PMID- 19566374 TI - Bonding effectiveness of different adhesion approaches to unground versus ground primary tooth enamel. AB - AIM: This study aims to evaluate the bonding effectiveness of self-etch and etch and-rinse adhesive systems in on intact and ground primary tooth enamel. STUDY DESIGN: Sixty primary incisors were divided into 6 groups according to the adhesive system (etch-and-rinse - Adper Single Bond 2 - SB, 2 steps self-etch Clearfil SE Bond - SE, and 1 step self-etch - One Up Bond F Plus OBF) and to the substrate (ground or intact enamel): G1-SB/intact enamel; G2-SE/intact enamel; G3 OBF/intact enamel; G4-SB/ground enamel; G5- SE/ground enamel and G6-OBF/ground enamel. METHODS: Microshear bond test specimens were prepared with microhybrid composite and after 24h of water storage the microshear test was performed. STATISTICS: Data were submitted to statistical analysis using two-way ANOVA and Tukey's tests (p<0.05). RESULTS: Comparing the enamel characteristics (ground or intact) only when SE was used a statistically significant difference was found, as G2 (21.12+/-4.52) was statistically lower than G5 (33.29+/-5.44). Among the intact enamel groups, SB (26.11+/-7.56) was statistically superior to SE (21.12+/ 4.52) and OBF (17.01+/-3.96). However, when comparisons were made among groups of ground enamel, SE (33.29+/-5.44) was significantly higher than SB (26.35+/-8.18) and OBF (17.52+/-3.46). CONCLUSION: The two-step self-etch adhesive system is a reliable alternative to etch and rinse adhesive systems on both ground and intact primary enamel. PMID- 19566375 TI - Dental development in children with cleft lip and palate: a comparison between unilateral and bilateral clefts. AB - AIM: The aims of this study were to assess and compare the dental development in patients with unilateral (UCLP) and bilateral (BCLP) cleft lip and palate. STUDY DESIGN: Dental developmental delay was compared between 40 UCLP patients (20 boys and 20 girls, average age 9.6 +/- 2.7 years) and 40 BCLP patients matched by sex and age (to the nearest 6 months). For the purpose of comparison, a total of 80 panoramic radiographs of unaffected group matching in age and sex were selected randomly. METHODS: Using panoramic radiographs, the seven left mandibular permanent teeth, second molar to central incisor, were rated on eight stage scales using the methods described by Demirjian et al. [1973]. The stage of each tooth was converted to the corresponding numeric value, and then all values were added to obtain a dental maturity score, which corresponded to a dental age. STATISTICS: The differences in developmental delay between the two cleft groups were determined using independent t-test. RESULTS: There was statistically significant difference (P<0.0001) between chronologic and dental age in cleft patients. The mean dental development delay in UCLP was 0.34 years and for BCLP was 0.61 years. BCLP patients were found to have a greater delay than UCLP patients (P<0.05). Girls showed an advanced dental development in both types of clefts. Comparison between the two cleft groups showed only significant difference in the amount of delay in female subjects. CONCLUSION: Dental development in UCLP and BCLP patients was significantly delayed compared to non cleft reference. Dental development in boys was significantly behind that of girls. A significant difference in the amount of delay between both cleft types was found only in female subjects. PMID- 19566376 TI - Management of unilateral condylar fracture in a 9.6-year-old female. AB - BACKGROUND: Fractures represent a particular kind of pathology in children, as they take place within a rapidly growing organism. Condylar fractures must be focused upon not only as a cause of direct damage to the bone structure, but also with regards to future disorders in the dentofacial development, skeletal deformities and articular dysfunctions. The treatment aims at restoring function, occlusion, and symmetry. Aim of this paper is to emphasise, through the description of a case report, the need for correct diagnosis and the importance of a functional treatment by means of early muscle activation, in order to prevent any consequences on facial development. CASE REPORT: A case of a 9.6-year old female with monocondylar and intracapsular fracture following a car accident is described. In accordance with the literature, the patient did not undergo surgical treatment, but followed a functional orthodontic protocol. The significant symptoms reduction and the rapid improvement of mandibular functions confirm the excellent healing potential in paediatric age, while the X-ray evaluations 1 year and 3.7 years later testify the condylar reshaping. Since the latter can continue over many years, the clinical and radiological follow-ups of the patient will have to cover the entire growth period. PMID- 19566377 TI - T-cell epitope analogues from carcinoembryonic antigen for vaccination against cancer: WO2009002418. AB - BACKGROUND: Cancer is one of the leading causes of death in the Western world. Therapeutic vaccination to target minimal residual disease or prevent tumor recurrence represents an interesting and novel alternative for treatment of tumor diseases. T-cell peptide epitopes are commonly used as vaccine candidates for the induction of antitumor immune responses. By modifying the amino-acid sequence of the peptide at certain, so-called anchor positions, the binding affinity to MHC class I and the immunogenicity of the peptide can be improved. Vaccination with the modified peptide analogue can then be used to induce an immune response to the wild-type epitope. METHOD: The present application concerns the use of peptides representing wild-type T-cell epitopes and analogues from carcinoembryonic antigen for vaccination against cancer. The stated claims also include the use of these epitopes in several other vaccine modalities, including RNA, DNA and adenoviral vector vaccines. CONCLUSION: Although the available data clearly support the basic claims that some of the peptide analogues indeed are able to induce a potent immune response in mice to the corresponding wild-type epitopes, the lack of in vivo antitumor data for any of the covered vaccine modalities prevents a thorough evaluation of the stated claims. PMID- 19566378 TI - Patented small molecules against psoriasis. AB - BACKGROUND: It is hypothesized that psoriasis is an autoimmune disease. The most recent therapeutic approach that proved to be more effective than earlier methods of treatment is the use of mAb/fusion proteins. Efforts nowadays are focused on investigating the antipsoriatic affect of small molecules that can be administered orally, some of which are capable of entering cells, and being selective in targeting intracellular pathways. OBJECTIVE: Preclinical patented small molecules that are recommended for the treatment of psoriasis are reviewed. Emphasis is placed on their mechanism of action. METHODS: http://ep.espacenet.com/ , Pubmed, Scopus and Google websites were the main sources used for the patented small molecule search. A number of patents were poorly described and difficulties were faced in trying to figure out the patentee(s) explanation. Moreover, most patents were recommended for the treatment of a number of autoimmune diseases and cancer, and not only for psoriasis. RESULTS/CONCLUSIONS: Small molecules that inhibit the activation of T lymphocytes, leukocyte trafficking, leukotriene activity/production and angiogenesis, and promote apoptosis have been patented. Small molecules that have been patented for the treatment of other autoimmune diseases and could be used for treating psoriasis are described. Moreover, other possible mechanistic approaches using small molecules are discussed. PMID- 19566379 TI - A cocaine body packer case report: clinical and forensic aspects. PMID- 19566380 TI - Mushroom poisoning from species of genus Inocybe (fiber head mushroom): a case series with exact species identification. AB - BACKGROUND: Many species of the genus Inocybe (family Cortinariaceae, higher Basidiomycetes) are muscarine-containing mycorrhizal mushrooms, ubiquitous around the world. The few published reports on the poisonous Inocybe mushrooms are often limited by the inadequate identification of the species. The clinical course of patients with typical muscarinic manifestations, in whom Inocybe spp. was unequivocally identified, is reported. CASE SERIES: Between November 2006 and January 2008 14 consecutive patients with typical muscarinic syndrome after mushroom ingestion were recorded. The clinical manifestations included combinations of nausea, vomiting, diarrhea, abdominal pain, hypersalivation, diaphoresis, tachycardia, bradycardia, hypotension, lacrimation, blurred vision, miosis, tremor, restlessness, flushing, and syncope. Time to onset of toxicity ranged between 15 min and 2 h after consumption, 5 h in one patient. Treatment was supportive, including intravenous fluids, antiemetics, and 1 mg atropine intravenously. Full recovery ensued within 12 h. In all the cases, an expert mycologist unequivocally identified the leftovers of the consumed mushrooms as Inocybe fastigiata, Inocybe geophylla, and Inocybe patouillardii. CONCLUSION: In this case series of patients who ingested identified muscarine-containing mushrooms supportive treatment and atropine resulted in recovery in all cases. PMID- 19566381 TI - Use of clonidine in the prevention and management of neonatal abstinence syndrome. AB - INTRODUCTION: Neonatal abstinence syndrome (NAS) is a complicated medical condition with treatment regimens that traditionally have included methadone and other opioids, barbiturates, and benzodiazepines. We describe a case series in which clonidine was used for the prevention and management of patients with NAS. PATIENTS AND METHODS: Medical records of infants treated with clonidine for NAS from January 2003 to March 2006 were reviewed for gestational age, birth weight, NAS score, dose of clonidine, duration of treatment, and additional medications required. RESULTS: Fourteen patients were identified. The mean gestational age was 30.1 weeks (range 24.4-40.7 weeks); three patients were full-term. Eleven had been on intravenous fentanyl for sedation; three were born to opioid-dependent mothers. All patients were treated with clonidine, administered in doses of 0.5 1.0 mcg/kg orally every 6 h. No patient received opioids. Mean duration of treatment was 6.8 days (range 4-15). Mean abstinence scores were 6.4 pretreatment (range 0-20) and 1.9 posttreatment (range 0-5). No patients suffered an adverse event (hypotension, bradycardia, excessive sedation, and oxygen desaturation) from clonidine administration, and no seizures were identified. CONCLUSIONS: Our data suggest that clonidine may be a reasonable alternative to more traditional agents used to prevent or treat NAS. We agree with the statement of the American Academy of Pediatrics Committee on Drugs that states that larger trials and pharmacologic data are needed before the routine use of clonidine can be recommended. PMID- 19566382 TI - The effect of olanzapine pretreatment on acute cocaine toxicity in mice. AB - BACKGROUND: Acute cocaine poisoning causes neuroexcitation and can be fatal. The toxic effects of cocaine can be attenuated by antagonists of serotonin, muscarinic cholinergic, and dopamine receptors. Olanzapine, an atypical antipsychotic medication, is an antagonist of these receptors. The objective of this study is to evaluate the efficacy of olanzapine pretreatment for attenuation of acute cocaine toxicity using a mouse model. METHODS: Eighty male CF-1 mice were randomly assigned to olanzapine (1 mg/kg) or placebo pretreatment. Fifteen minutes later, all animals received 103 mg/kg intraperitoneal cocaine. RESULTS: Overall mortality was 11% for olanzapine-treated animals and 45% for placebo. Olanzapine also appeared to alter the characteristics of seizures due to cocaine. CONCLUSIONS: In this model of acute cocaine toxicity, olanzapine pretreatment attenuated acute cocaine toxicity. Olanzapine should be evaluated further as a potential treatment for acute cocaine poisoning. PMID- 19566383 TI - Limiting paracetamol pack size: has it worked in the UK? AB - INTRODUCTION: Paracetamol poisoning is a major health problem worldwide. Limitation of pack size is an approach increasingly advocated to reduce rates of suicide and serious self-harm from this agent. The United Kingdom adopted such a policy in 1998, restricting non-pharmacy sales to 8 g and pharmacy to pack sizes of 16 g. METHODS: A literature review was conducted and commentary written on the impact of the change in the United Kingdom on a variety of indices of paracetamol overdose. RESULTS: Potential markers of effect identified included paracetamol sales, poisons information data, laboratory results, liver unit referrals, and hospital activity and mortality data. Initial reports suggested effects associated with the legislation, but longer term suicide trend analysis has not confirmed these early findings, which were confounded by population trends in self-harm. CONCLUSION: Paracetamol pack size limitation as applied in the United Kingdom has not reduced paracetamol-related death. Reasons postulated for this failure include patient avoidance of the legislation's intentions, patient confusion, and ineffectiveness of the regulations as conceived and implemented. PMID- 19566385 TI - Real-time surveillance of illicit drug overdoses using poison center data. AB - BACKGROUND: In early 2006, government and media sources reported that crime syndicates were mixing fentanyl with heroin. This was followed by an increase in heroin overdoses and opiate-related deaths. The most recent fentanyl outbreak illustrated the need for identifying and establishing effective and responsive real-time surveillance tools to monitor drug overdoses in the United States. OBJECTIVE: In this study, poison call center data from Illinois were evaluated to determine whether the data could have detected the outbreak that occurred in Illinois in early 2006 and whether it could be used for real-time surveillance. METHODS: For this analysis, a two-step approach was used to analyze potential heroin-related calls. First, the data were analyzed retrospectively to identify whether any significant temporal shifts occurred, then a prospective analysis was conducted to simulate real-time surveillance. RESULTS: Between 2002 and 2007, there were a total of 1,565 potential heroin-related calls, and the calls increased by 63.6% in 2006 compared to 2005. In the prospective analysis, the principal model would have identified the outbreak in March 2006. CONCLUSIONS: If there had been a real-time surveillance program using poison center data, the outbreak would have been identified 1 month before the initial postmortem reports to the Centers for Disease Control and Prevention at the end of April 2006. Poison center data provide the potential for an earlier warning system than postmortem data sources, because the reports are usually made within hours of the exposure. Poison center data can be effectively used to monitor heroin-related exposures. PMID- 19566387 TI - Flow, compressive, and bioadhesive properties of various blends of poly(ethylene oxide). AB - BACKGROUND: Poly (ethylene oxide) (PEO) is nonionic, water soluble, and highly hydrophilic polymer with well-established applications in mucoadhesives, water soluble films, rheology control agents and thickeners, and additives in pharmaceutical products. METHODS: Different powder blends containing PEO in varying proportions were evaluated for their flow, compressive, and bioadhesive properties and subsequently compressed into gastroretentive tablets. Two optimized formulations, on the basis of above-mentioned examinations, were subjected to gamma scintigraphy studies on human volunteers. RESULTS: The values of bulk and tapped densities, Hausner ratio and Carr index, angle of repose, loss on drying, total moisture content, and particle size distribution provided a fine estimation of flowability and compressibility of the powder blends. Further, apart from the routine pharmacopoeial assessments, the evaluation of compressed tablets for their surface pH in both acidic and basic environments nullified the possibility of any irritation to the membrane where it is intended to adhere. The measurement of swelling index and bioadhesive strength of tablets revealed that both the parameters were a direct function of the concentration of PEO in the tablet. The results of gamma scintigraphy indicated a fourfold increase in the gastric retention time of the optimized formulation vis-a-vis control formulation. CONCLUSION: The results indicate that PEO, in a concentration of 10 50% (w/w), can be successfully employed in manufacturing gastroretentive tablets. PMID- 19566394 TI - Ultrasonic atomization and subsequent desolvation for monoclonal antibody (mAb) to the glycoprotein (GP) IIIa receptor into drug eluting stent. AB - An eluting-stent system with mAb dispersed in the PLLA (poly (L-lactic acid)) was validated in vitro. Specifically designed spray equipment based on the principle of ultrasonic atomization was used to produce a thin continuous PLLA (poly (L lactic acid)) polymer coating incorporating monoclonal antibody (mAb). This PLLA coating was observed in light microscopy (LM) and scanning electron microscopy (SEM). The concentration of the monoclonal antibody (mAb) to the platelet glycoprotein (GP) IIIa receptor and the eluting rate were then measured by a radioisotope technique with (125)I-labelled GP IIIa mAb. An in vitro perfusion circuit was designed to evaluate the release rates at different velocities (10 or 20 ml min(-1)). The PLLA coating was thin and transparent, uniformly distributed on the surface of the stent. Three factors influenced its thickness: PLLA concentration, duration and gas pressure. The concentration of mAb was influenced by the duration of absorption and the concentration of the mAb solution; the maximum was 1662.23 + or - 38.83 ng. The eluting rate was fast for the first 2 h, then decreased slowly and attained 80% after 2 weeks. This ultrasonic atomization spray equipment and technological process to prepare protein eluting-stents were proved to be effective and reliable. PMID- 19566395 TI - Chitosan derivative nanocarrier: safety evaluation,antibacterial property and ascorbyl palmitate encapsulation. AB - A chitosan derivative, methyl ether-terminated poly(ethylene oxide)-4 methoxycinnamolyphthaloylchitosan (PCPLC) was prepared, characterized and self assembled into nanoparticles. Encapsulation of ascorbyl palmitate (AP) into PCPLC gave 6890.98 nm particles with encapsulation efficiency of 84% at 56% drug loading. The encapsulated AP showed significant improved stability as examined by 1H NMR spectroscopy.The obtained particles displayed no short-term cytotoxicity against the human skin melanoma A-375 cell line using the MTT assay and no short term skin irritation on human volunteers using a single topical application as patch and photopatch tests. In addition, aqueous suspension of PCPLC nanoparticles successfully inhibited the growth of Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923. PMID- 19566396 TI - Mobile phone-based telemedicine system for the home follow-up of patients undergoing ambulatory surgery. AB - A pilot study was done to address the efficacy of a General Packet Radio Service mobile phone-based telemedicine system used to improve follow-up after ambulatory surgery. The method involves sending images of surgical wounds or other areas from the patient's home, to assess local complications and avoid unnecessary hospital visits. Ninety-six (N = 96) patients were enrolled in the study. The phone used was a Nokia 6600, which provides images in Joint Photographic Experts Group format. These images were sent via e-mail and visualized on a standard 17 inch screen of a personal computer. After the follow-up period, self-reported patient satisfaction was assessed by analyzing the replies to a 9-item questionnaire. Thirty of the 96 patients (31.3%) reported local problems including: hematoma in 20 (66.7%) patients, surgical bandage blood-stained in 7 (23.3%), exudates in 1 (3.3%), allergic skin reactions in 1 (3.3%), and bandage too tight in 1 (3.3%). In total, 225 photographs were evaluated by 3 physicians. In all cases, it was possible to identify and assess the postoperative problem with consensus among the 3 physicians. Images served to resolve patients' concerns in 20 individuals (66.7%). In 10 patients (33.3%), concerns were satisfied but it was suggested that follow-up images be sent in the following days. Only 1 patient (3.3%) was asked to visit the hospital. The telemedicine system proposed increases the efficiency of home follow-up to ambulatory surgery, avoids unnecessary hospital visits, and clearly improves patient satisfaction. PMID- 19566397 TI - Pre-experience perceptions about telemedicine among African Americans and Latinos in South Central Los Angeles. AB - This study explores perceptions about telemedicine among urban underserved African American and Latino populations. Telemedicine has been advanced as a vehicle to increase access to specialty care among the urban underserved, yet little is known about its acceptability among these populations. We conducted 10 focus groups with African American and Latino participants (n = 87) in urban Los Angeles in order to explore perceptions about this novel type of care. We found that concerns about telemedicine varied between the two racial/ethnic groups. These findings have implications for important issues such as adoption of telemedicine, patient satisfaction, and doctor-patient interaction. It will be critical to consider perceptions of this healthcare innovation in the development of strategies to market and implement telemedicine among urban, underserved African American and Latino populations. PMID- 19566400 TI - Innovative programs in telemedicine HealthBridge. PMID- 19566399 TI - Acute asthma management by a pediatric after-hours call center. AB - To describe an asthma management protocol used in a nurse-staffed pediatric After Hours Call Center (AHCC) that incorporates severity-based home treatment recommendations and follow-up call assessments. Call records for asthma advice from January 1, 2004 to June 30, 2004 were identified retrospectively and reviewed. Descriptive statistics were used to report patient demographics, frequencies of symptom severity zones (Red, Yellow, or Green) at initial calls, frequencies of call dispositions designating care advice provided (including home treatment recommendations and seeking emergency department [ED] care), and changes in severity zones between initial calls and follow-up calls when nurses reassessed patients after recommended home treatment. During the study period, 3,632 asthma calls (2,439 initial; 1,193 follow-up) were managed by AHCC nurses. Initial calls were classified mostly as Red (28%) or Yellow (42%) severity zones; 27% were Green zone and 3% could not be categorized. Fifty-two percent of initial calls with Red or Yellow severity zones involved home treatment recommendations; 50% of those Red zone and 63% of those Yellow zone calls had improved severity zones at follow-up call assessments. Twenty-eight percent of patients with home treatment recommendations were referred to the ED at the time of follow-up call nurse reassessment. This telephone-based nurse-staffed pediatric acute asthma management protocol includes provision of severity-based home treatment recommendations and follow-up assessments, and improved symptoms for many children with acute exacerbations. This protocol may also be successful in other locations and may improve outcomes, such as reduction in ED visits. PMID- 19566401 TI - Adherence among telemonitored patients with heart failure to pharmacological and nonpharmacological recommendations. AB - In many healthcare systems, the outcomes of care for patients with heart failure (HF) are in need of improvement. By applying telemonitoring, this study aimed to assess its short-term impact on patients' disease-specific knowledge, adherence, and depression. As part of a larger trial (N = 1,023, 17 centers), this study reports preliminary findings among 101 patients from three Dutch hospitals. Patients were randomized to receive care using telemonitoring or standard care. Data concerning patients' disease-specific knowledge, adherence to pharmacological and nonpharmacological recommendations, and depression were collected by postal questionnaires. In this study, data collected before randomization and 3 months afterwards were analyzed. Disease-specific knowledge improved significantly in two of the three hospitals (p < 0.001 and p = 0.040). Adherence in terms of fluid restrictions (p = 0.012), daily weighing (p < 0.001), physical exercising (p = 0.034), and alcohol restrictions (p = 0.040) improved significantly in the telemonitoring group. Additionally, in contrast with the hypothesized increase of depression, the use of telemonitoring resulted in a substantial but statistically not significant decrease in depression. The improved adherence rates within the 3-month study period underscore the potential of telemonitoring to enhance self-management among HF patients and consequently its potential impact on other outcomes. Longer-term results will enable solid conclusions to be reached concerning the relation between telemonitoring and patients' adherence. PMID- 19566403 TI - The Fourteenth Annual International Meeting and Exposition of the American Telemedicine Association. April 26-28, 2009 Las Vegas, NV. PMID- 19566405 TI - Is this a defining moment? PMID- 19566402 TI - American Telemedicine Association 2009 presidential address. PMID- 19566406 TI - Telemedicine in emergency neurological service provision in Singapore: using technology to overcome limitations. AB - Medical emergencies form a significant proportion of neurological and neurosurgical practice. The highly specialized nature of neurology and neurosurgery limits expertise to a small number of highly specialized centers; hence, many neurological emergencies will be managed in regional hospitals without comprehensive support. The common theme in neurological emergencies lies with prompt diagnosis, rapid access to scans, accurate scan interpretation, and quick institution of appropriate treatment. Failure in any one of these can result in devastating and permanent neurological deficits or death. The judicious use of information and communication technology can overcome some of the limitations imposed by scarce resources. Multimedia messaging service can be used to transmit important scan images to experienced staff to faciliate accurate and prompt diagnosis and commence optimal treatment. Telestroke based on the remote evaluation of acute ishemic stroke model can also be utilized so that selected stroke patients can be given intravenous tissue plasminogen activator in the emergency department of a regional hospital with the supervision of a stroke neurologist. We describe our experience with these technological initiatives to improve clinical care and outcome in our patients. PMID- 19566407 TI - Motion-based angiogenesis analysis: a simple method to quantify blood vessel growth. AB - Existing methods to quantify angiogenesis range from image analysis of photographs to fluorescent microscopy. These methods are often time consuming and costly; they also may not detect capillaries if they are indistinct from the background of the image. We have developed a simple method based on the motion of blood to create an image that reveals the entire angiogenic vasculature. Two image analysis software programs were used separately to demonstrate the method. Using either ImageJ or Environment for Visualizing Images, we analyzed a video clip of regenerated tissue from the partially amputated caudal fin of a zebrafish (Danio rerio). The deviations among the frames in the video stack were calculated to reveal pixels where motion has occurred. The resulting image highlighted all vessels through which blood flowed and allowed for automatic quantification of the newly developed vasculature. Using this method, we quantified the angiogenic action of basic fibroblast growth factor and vascular endothelial growth factor, as well as suppression of angiogenesis by an inhibitor. In a preliminary study, we also found that it could be used to trace the developing vasculature in zebrafish embryos. Thus, motion-based angiogenesis analysis may provide an easy and accurate quantification of angiogenesis. PMID- 19566408 TI - Growth and survival of zebrafish (Danio rerio) fed different commercial and laboratory diets. AB - The need to develop standardized diets to support zebrafish (Danio rerio) research is supported by the knowledge that specific dietary ingredients, nutrients, or antinutritional factors in diets have been shown to affect development and growth of adult D. rerio and their offspring. In this study, there were seven dietary treatments consisting of five commercially available diets and two laboratory-prepared diets, three replicates per treatment. Fish were fed ad libitum twice daily for 9 weeks. At 9 weeks, both weight and length were recorded to determine condition indices. D. rerio fed one of the laboratory prepared diets had significantly higher weights than individuals fed any of the other diets and exhibited significantly higher lengths than those fed five of the six remaining diets. Although there were significant differences in general growth demographics (length/weight) after the 9-week feeding trial, no significant differences in overall health of D. rerio were observed for the different dietary treatments as determined by statistical analysis of condition factor indices (K = [weight x 100]/length(3)). The success achieved with the laboratory-prepared diets represents the foundation for establishing an open formulation nutritional standard to ensure that the D. rerio model for research does not generate confounding research results caused by nutritional vagaries. PMID- 19566409 TI - Before maternal-zygotic transition ... There was morphogenetic function of nuclei. AB - The idea of a major developmental transition that includes the activation of the embryonic genome has a long history. In the 1950-1960s this concept was developed to a large extent due to the efforts of Alexander Neyfakh, who described a specific type of deleterious effect resulting from X-ray irradiation of fish eggs. He interpreted the radiation-sensitive target as the nucleus and established the onset of the function of the zygotic genome, naming it the morphogenetic function of nuclei, what we call now the midblastula transition. Most of his studies were performed using the loach (Misgurnus fossilis), a European teleost. Neyfakh's efforts paved the way to understanding the whole phenomenon of the maternal-zygotic transition. PMID- 19566410 TI - Management of chronic unilateral hematuria by ureterorenoscopy. AB - BACKGROUND AND PURPOSE: Chronic unilateral hematuria is characterized by intermittent or continuous gross hematuria that cannot be diagnosed using standard radiology and hematology methods. In the past, it was managed with partial or total nephrectomy. In the age of minimally invasive procedures, however, endoscopy has enabled more accurate diagnosis and management. We analyzed our experience with transurethral ureterorenoscopy using a flexible ureteroscope to determine the feasibility and success of endoscopic management of renal hematuria. PATIENTS AND METHODS: We reviewed the records of 13 patients who presented with chronic unilateral hematuria, in whom radiologic and laboratory tests failed to reveal the source of bleeding. In the cases in which the lesion was identified, after complete inspection of the collecting systems, the bleeding site was treated ureteroscopically with a holmium: yttrium-aluminum-garnet (YAG) laser. RESULTS: Follow-up ranged from 4 to 60 months (mean 26 mos). During the follow-up of the 13 patients, 11 remained symptom-free, with only one session of flexible ureterorenoscopy necessary. Relapse occurred in two patients after 4 months and 6 months, respectively; during a second session of flexible ureteroscopy, the bleeding site was successfully identified and cauterized with a holmium:YAG laser. No surgical complications occurred. CONCLUSIONS: Conservative treatment of patients with chronic unilateral hematuria should always be considered. Laser ureteroscopic treatment is an excellent method and should be considered as the first option for the management of chronic unilateral hematuria. PMID- 19566411 TI - Laparoendoscopic single-site surgery: early experience with tumor nephrectomy. AB - BACKGROUND AND PURPOSE: Laparoendoscopic single-site surgery (LESS) represents the closest surgical technique to scar-free surgery. We performed LESS for renal tumor nephrectomy in eight patients to assess feasibility and perioperative outcome. PATIENTS AND METHODS: Eight patients with a body mass index (BMI)